PMID- 10818011 TI - Doctors demand more information on GMC candidates. General Medical Council. PMID- 10818013 TI - Surgeon dies from hospital exposure to asbestos PMID- 10818012 TI - Controversy over new editor at New England Journal of Medicine. PMID- 10818014 TI - In brief PMID- 10818016 TI - Health websites in US propose new ethics code. PMID- 10818015 TI - Doctors' arrogance blamed for retention of childrens' organs. PMID- 10818018 TI - Cochlear hair cells generated in mammals after birth PMID- 10818017 TI - Coverage for cancer treatment extended to poor women in US. PMID- 10818019 TI - Too early to introduce colorectal screening, say specialists PMID- 10818020 TI - Heart transplants suspended in Scotland. PMID- 10818021 TI - Doctor puts herring on prescription. PMID- 10818022 TI - WHO accused of stifling debate about infant feeding. PMID- 10818023 TI - Drug industry is unwilling to run trials in children. PMID- 10818024 TI - Role of vaccinations as risk factors for ill health in veterans of the Gulf war: cross sectional study. AB - OBJECTIVES: To explore the relation between ill health after the Gulf war and vaccines received before or during the conflict. To test the hypothesis that such ill health is limited to military personnel who received multiple vaccines during deployment and that pesticide use modifies any effect. DESIGN: Cross sectional study of Gulf war veterans followed for six to eight years after deployment. SETTING: UK armed forces. PARTICIPANTS: Military personnel who served in the Gulf and who still had their vaccine records. MAIN OUTCOME MEASURES: Multisymptom illness as classified by the Centers for Disease Control and Prevention; fatigue; psychological distress; post-traumatic stress reaction; health perception; and physical functioning. RESULTS: The response rate for the original survey was 70.4% (n=3284). Of these, 28% (923) had vaccine records. Receipt of multiple vaccines before deployment was associated with only one of the six health outcomes (post-traumatic stress reaction). By contrast five of the six outcomes (all but post-traumatic stress reaction) were associated with multiple vaccines received during deployment. The strongest association was for the multisymptom illness (odds ratio 5.0; 95% confidence interval 2.5 to 9.8). CONCLUSION: Among veterans of the Gulf war there is a specific relation between multiple vaccinations given during deployment and later ill health. Multiple vaccinations in themselves do not seem to be harmful but combined with the "stress" of deployment they may be associated with adverse health outcomes. These results imply that every effort should be made to maintain routine vaccines during peacetime. PMID- 10818025 TI - Meta-analysis of increased dose of inhaled steroid or addition of salmeterol in symptomatic asthma (MIASMA). AB - OBJECTIVE: To examine the benefits of adding salmeterol compared with increasing dose of inhaled corticosteroids. DESIGN: Systematic review of randomised, double blind clinical trials. Independent data extraction and validation with summary data from study reports and manuscripts. Fixed and random effects analyses. SETTING: EMBASE, Medline, and GlaxoWellcome internal clinical study registers. MAIN OUTCOME MEASURES: Efficacy and exacerbations. RESULTS: Among 2055 trials of treatment with salmeterol, there were nine parallel group trials of >/=12 weeks with 3685 symptomatic patients aged >/=12 years taking inhaled steroid in primary or secondary care. Compared with response to increased steroids, in patients receiving salmeterol morning peak expiratory flow was greater at three months (difference 22.4 (95% confidence interval 15.0 to 30.0) litre/min, P<0.001) and six months (27.7 (19.0 to 36.4) litre/min, P<0.001). Forced expiratory volume in one second (FEV(1)) was also increased at three months (0.10 (0.04 to 0.16) litres, P<0.001) and six months (0.08 (0.02 to 0.14) litres, P<0.01), as were mean percentage of days and nights without symptoms (three months: days-12% (9% to 15%), nights-5% (3% to 7%); six months: days-15% (12% to 18%), nights-5% (3% to 7%); all P<0.001) and mean percentage of days and nights without need for rescue treatment (three months: days-17% (14% to 20%), nights-9% (7% to 11%); six months: days-20% (17 to 23%), nights-8% (6% to 11%); all P<0.001). Fewer patients experienced any exacerbation with salmeterol (difference 2.73% (0.43% to 5.04%), P=0. 02), and the proportion of patients with moderate or severe exacerbations was also lower (2.42% (0.24% to 4.60%), P=0.03). CONCLUSIONS: Addition of salmeterol in symptomatic patients aged 12 and over on low to moderate doses of inhaled steroid gives improved lung function and increased number of days and nights without symptoms or need for rescue treatment with no increase in exacerbations of any severity. PMID- 10818026 TI - Cost effectiveness of an intensive blood glucose control policy in patients with type 2 diabetes: economic analysis alongside randomised controlled trial (UKPDS 41). United Kingdom Prospective Diabetes Study Group. AB - OBJECTIVE: To estimate the cost effectiveness of conventional versus intensive blood glucose control in patients with type 2 diabetes. DESIGN: Incremental cost effectiveness analysis alongside randomised controlled trial. SETTING: 23 UK hospital clinic based study centres. PARTICIPANTS: 3867 patients with newly diagnosed type 2 diabetes (mean age 53 years). INTERVENTIONS: Conventional (primarily diet) glucose control policy versus intensive control policy with a sulphonylurea or insulin. MAIN OUTCOME MEASURES: Incremental cost per event-free year gained within the trial period. RESULTS: Intensive glucose control increased trial treatment costs by pound 695 (95% confidence interval pound 555 to pound 836) per patient but reduced the cost of complications by pound 957 (pound 233 to pound 1681) compared with conventional management. If standard practice visit patterns were assumed rather than trial conditions, the incremental cost of intensive management was pound 478 (-pound 275 to pound 1232) per patient. The within trial event-free time gained in the intensive group was 0.60 (0.12 to 1.10) years and the lifetime gain 1.14 (0.69 to 1.61) years. The incremental cost per event-free year gained was pound 1166 (costs and effects discounted at 6% a year) and pound 563 (costs discounted at 6% a year and effects not discounted). CONCLUSIONS: Intensive blood glucose control in patients with type 2 diabetes significantly increased treatment costs but substantially reduced the cost of complications and increased the time free of complications. PMID- 10818027 TI - Effect of beer drinking on risk of myocardial infarction: population based case control study. PMID- 10818029 TI - Bladders and brobdingnag PMID- 10818028 TI - The war in south africa: the hot dry summer PMID- 10818030 TI - The impact of patients' preferences on the treatment of atrial fibrillation: observational study of patient based decision analysis. AB - OBJECTIVE: To investigate the impact of patients' preferences for the treatment of atrial fibrillation, by using individualised decision analysis combining probability and utility assessments into a decision tree. DESIGN: Observational study based on interviews with patients. SETTING: Eight general practices in Avon. PARTICIPANTS: 260 randomly selected patients aged 70-85 years with atrial fibrillation. MAIN OUTCOME MEASURES: Patients' treatment preferences regarding anticoagulation treatment (warfarin) after individualised decision analysis; comparison of these preferences with treatment guidelines on the basis of comorbidity and absolute risk and compared with current prescription. RESULTS: Of 195 eligible patients, 97 participated in decision making using decision analysis. Among these 97, the decision analysis indicated that 59 (61%; 95% confidence interval 50% to 71%) would prefer anticoagulation treatment considerably fewer than those who would be recommended treatment according to guidelines. There was marked disagreement between the decision analysis and guideline recommendations (kappa=0.25 or less). Of 38 patients whose decision analysis indicated a preference for anticoagulation, 17 (45%) were being prescribed warfarin; on the other hand, 28 (47%) of 59 patients were not being prescribed warfarin although the results of their decision analysis suggested they wanted to be. CONCLUSIONS: In the context of shared decision making, individualised decision analysis is valuable in a sizeable proportion of elderly patients with atrial fibrillation. Taking account of patients' preferences would lead to fewer prescriptions for warfarin than under published guideline recommendations. Decision analysis as a shared decision making tool should be evaluated in a randomised controlled trial. PMID- 10818031 TI - Recent advances in rehabilitation. PMID- 10818033 TI - When medical journals were much less boring PMID- 10818034 TI - Eavesdropping on the great and the good PMID- 10818032 TI - Lesson of the week. Tinea capitis in adults. PMID- 10818036 TI - Ophthalmology in spain PMID- 10818035 TI - ABC of arterial and venous disease. Varicose veins. PMID- 10818037 TI - Health impact assessment. PMID- 10818039 TI - Obituaries PMID- 10818038 TI - Modernizing the NHS. Prevention and the reduction of health inequalities. PMID- 10818040 TI - GMC will get help to deal with more cases PMID- 10818041 TI - Fevered lives: tuberculosis in american culture since 1870 PMID- 10818042 TI - The museum of emotions PMID- 10818043 TI - Bookcase PMID- 10818044 TI - The steady drip of biased reporting PMID- 10818046 TI - Failures of process at the GMC PMID- 10818045 TI - Gulf war syndrome PMID- 10818047 TI - Happy forms? PMID- 10818049 TI - Multiple vaccines received during deployment may explain gulf war illness PMID- 10818048 TI - The gap between expectations and reality PMID- 10818050 TI - Adding salmeterol is more effective than increasing inhaled steroids in symptomatic asthma PMID- 10818051 TI - Tight glucose control is cost effective in diabetes PMID- 10818052 TI - Beer drinking protects against heart disease PMID- 10818054 TI - All policies should be subjected to some form of health impact assessment PMID- 10818053 TI - Patient preferences conflict with guidelines on atrial fibrillation PMID- 10818055 TI - Recognition of systolic hypertension for hypertension. PMID- 10818056 TI - Clinical Advisory Statement. Importance of systolic blood pressure in older Americans. PMID- 10818057 TI - Hypokalemia associated with diuretic use and cardiovascular events in the Systolic Hypertension in the Elderly Program. AB - The treatment of hypertension with high-dose thiazide diuretics results in potassium depletion and a limited benefit for preventing coronary events. The clinical relevance of hypokalemia associated with low-dose diuretics has not been assessed. To determine whether hypokalemia that occurs with low-dose diuretics is associated with a reduced benefit on cardiovascular events, we analyzed data of 4126 participants in the Systolic Hypertension in the Elderly Program (SHEP), a 5 year randomized, placebo-controlled clinical trial of chlorthalidone-based treatment of isolated systolic hypertension in older persons. After 1 year of treatment, 7.2% of the participants randomized to active treatment had a serum potassium <3.5 mmol/L compared with 1% of the participants randomized to placebo (P<0.001). During the 4 years after the first annual visit, 451 participants experienced a cardiovascular event, 215 experienced a coronary event, 177 experienced stroke, and 323 died. After adjustment for known risk factors and study drug dose, the participants who received active treatment and who experienced hypokalemia had a similar risk of cardiovascular events, coronary events, and stroke as those randomized to placebo. Within the active treatment group, the risk of these events was 51%, 55%, and 72% lower, respectively, among those who had normal serum potassium levels compared with those who experienced hypokalemia (P<0.05). The participants who had hypokalemia after 1 year of treatment with a low-dose diuretic did not experience the reduction in cardiovascular events achieved among those who did not have hypokalemia. PMID- 10818058 TI - Diuretic treatment of systolic hypertension in the elderly. PMID- 10818059 TI - Skip patterns in DINAMAP-measured blood pressure in 3 epidemiological studies. AB - Blood pressure measured by the oscillometric, automated device DINAMAP in 3 large population-based studies sponsored by the National Heart, Lung, and Blood Institute (The Atherosclerosis Risk in Communities Study, The Family Heart Study, and the Hypertension Genetic Epidemiology Network Study) were reviewed to determine an apparent skip pattern in the measurement values. Across the 3 studies, 2 different DINAMAP models were evaluated on >350 000 different blood pressure measurements. Measurements were taken in various positions, on both arm and ankles, and under various conditions (eg, resting and during stress). The following systolic blood pressure values were consistently skipped by the device: 89, 119, 120, 124, 125, 130, 140, 141, 150, 160, 170, 180, 190, and 200 mm Hg. No skip pattern was detected for diastolic blood pressure. Pulse data, which were only available in the Hypertension Genetic Epidemiology Network Study, also showed the following skipped values: 95, 99, 103, 106, and 109 bpm. Consultation with the manufacturer, the Critikon Corporation, indicated that the use of an algorithm designed to improve the accuracy of the DINAMAP device prevents these values from being displayed. Assessment of the extent and direction of bias caused by the skipped values is difficult, given that the algorithm is proprietary. While the implications of the skipped values are not clear, it is important for clinicians and researchers to be aware of this feature. PMID- 10818060 TI - Blood pressure-measuring devices: time to open Pandora's box and regulate. PMID- 10818061 TI - Influence of diabetes and type of hypertension on response to antihypertensive treatment. AB - The aim of our investigation was to determine whether the presence of additional risk factors or type of hypertension (diastolic or isolated systolic) influences blood pressure (BP) response to treatment. The International Nifedipine GITS Study: Intervention as a Goal in Hypertension Treatment (INSIGHT) study is a double-blinded outcome comparison of calcium channel blockade with diuretics in high-risk patients aged 55 to 80 years. Dynamic randomization between nifedipine once daily and hydrochlorothiazide/amiloride was performed to ensure that approximately equal numbers of patients in the 2 groups had each of the major cardiovascular risk factors. Patients with isolated systolic hypertension were also separately randomized. Atenolol or enalapril was the mandatory second-line drug. In 5669 patients who completed the 18-week titration, BP fell from 172+/ 15/99+/-9 mm Hg (mean+/-SD) while receiving placebo to 139+/-12/82+/-7 mm Hg. Twenty-six percent of patients required 2 drugs, and 4% required 3 drugs. Patients with diabetes were the most resistant to treatment, requiring second and third drugs 40% and 100% more frequently than patients without diabetes and achieving marginally the highest final BP, for any risk group, of 141+/-13/82+/-8 mm Hg. Age, smoking, gender, hypercholesterolemia, left ventricular hypertrophy, and existing atherosclerosis had little (<1 mm Hg) or no influence on BP at the end of titration, but all except smoking slightly reduced the initial response of either systolic or diastolic BP. Patients with isolated systolic hypertension were slightly more responsive than average to treatment. Our findings suggest that in patients at high absolute risk of cardiovascular complications from hypertension, the risk factors themselves do not prevent the recommended BP targets from being achieved. PMID- 10818062 TI - Insulin-induced decrease in large artery stiffness is impaired in uncomplicated type 1 diabetes mellitus. AB - Normal insulin action in vivo involves a decrease in stiffness of large arteries (a decrease in aortic pressure augmentation). We determined whether the ability of insulin to decrease arterial stiffness is altered in uncomplicated type 1 diabetes. Nine type 1 diabetic men (age 28+/-2 years, body mass index 24+/-1 kg/m(2)) and 9 matched normal men were studied under normoglycemic hyperinsulinemic (sequential 2-hour insulin infusions of 1 [step 1] and 2 [step 2] mU x kg(-1) x min(-1)) conditions. Central aortic pressure waveforms were synthesized from those recorded in periphery with applanation tonometry on the radial artery and a validated reverse transfer function to construct the central aortic pressure wave every 30 minutes. This allowed the determination of aortic augmentation (the pressure difference between the first and the second systolic peaks) and the augmentation index (augmentation divided by pulse pressure), as the measure of stiffness of large arteries. Whole-body glucose uptake was 44% (step 1) and 37% (step 2) lower (P<0.001) in the diabetic patients than in the normal subjects. At baseline, before the insulin infusion, augmentation averaged 0+/-1 and 2+/-1 mm Hg (NS) and the augmentation index was -1.5+/-4.5% and 4.0+/ 3.7% (NS) in the normal and diabetic subjects, respectively. After 1 hour of hyperinsulinemia, the augmentation index had decreased significantly (P<0.01) to 9.5+/-4.8% in the normal subjects but remained at 4.4+/-4.2% in the diabetic patients. A significant decrease was not observed in the diabetic patients until 150 minutes (-1.2+/-4.1%, P<0.05 versus baseline). Whole-body glucose uptake was significantly inversely correlated with the change in the augmentation index during step 1 (r=-0.61, P<0.01). Insulin resistance in type 1 diabetes involves a defect in the ability of insulin to decrease central aortic pressure. This defect could predispose these patients to premature stiffening of large arteries. PMID- 10818063 TI - Intrinsic stiffness of the carotid arterial wall material in essential hypertensives. AB - We have previously shown that the decrease in large artery distensibility observed in patients with essential hypertension (HT group) was primarily due to an increase in distending pressure and not to hypertension-associated structural modifications of the artery, suggesting a functional adaptation of the wall material. To evaluate the elastic properties of the wall material of the common carotid artery, we determined Young's incremental elastic modulus (Einc) in the HT group and in normotensive subjects (NT group) as a function of blood pressure and circumferential wall stress. In 102 HT patients with never-treated essential hypertension and 40 age- and gender-matched NT subjects, the Einc-pressure and Einc-stress curves were calculated from intima-media thickness and from diameter and pressure waveforms, determined with echo tracking and aplanation tonometry, respectively. The "effective" stiffness of the wall material, determined through Einc calculated at mean blood pressure, was significantly higher in the HT than in the NT group. The "intrinsic" stiffness of the wall material, determined through Einc calculated at a common circumferential wall stress, did not differ between the 2 groups. However, when each group (HT and NT) was analyzed according to tertiles of age, the "intrinsic" stiffness of the arterial wall material was increased only in younger HT patients. In middle-aged and older HT patients, the intrinsic mechanical properties of the carotid arterial wall material were unchanged, and the increased stiffness of the common carotid artery in the HT group was due primarily to the increased level of blood pressure. These results also indicate that the deleterious effects of aging and hypertension on "intrinsic" stiffness are not additive. PMID- 10818064 TI - Vascular NADH/NADPH oxidase is involved in enhanced superoxide production in spontaneously hypertensive rats. AB - This study was designed to test the hypothesis that stimulation of nicotinamide adenine dinucleotide/nicotinamide adenine dinucleotide phosphate (NADH/NADPH) oxidase is involved in increased vascular superoxide anion (*O(2)(-)) production in spontaneously hypertensive rats (SHR). The study was performed in 16-week-old and 30-week-old normotensive Wistar-Kyoto rats (WKY(16) and WKY(30), respectively) and in 16-week-old and 30-week-old SHR (SHR(16) and SHR(30), respectively). In addition, 16-week-old SHR were treated with oral irbesartan (average dose 20 mg/kg per day) for 14 weeks (SHR(30)-I). Aortic NADH/NADPH oxidase activity was determined by use of chemiluminescence with lucigenin. The expression of p22phox messenger RNA was assessed by competitive reverse transcription-polymerase chain reaction. Vascular responses to acetylcholine were determined by isometric tension studies. Aortic wall structure was studied, determining the media thickness and the cross-sectional area by morphometric analysis. Whereas systolic blood pressure was significantly increased in the 2 groups of hypertensive animals compared with their normotensive controls, no differences were observed in systolic blood pressure between SHR(30) and SHR(16). No other differences in the parameters measured were found between WKY(16) and SHR(16). In SHR(30) compared with WKY(30), we found significantly greater p22phox mRNA level, NADH/NADPH-driven *O(2)(-) production, media thickness, and cross sectional area and an impaired vasodilation in response to acetylcholine. Treated SHR had similar NADH/NADPH oxidase activity and p22phox expression as the WKY(30) group. The vascular functional and morphological parameters were improved in SHR(30)-I. These findings suggest that an association exists between p22phox gene overexpression and NADH/NADPH overactivity in the aortas of adult SHR. Enhanced NADH/NADPH oxidase-dependent *O(2)(-) production may contribute to endothelial dysfunction and vascular hypertrophy in this genetic model of hypertension. PMID- 10818065 TI - Inversion of the intracellular Na(+)/K(+) ratio blocks apoptosis in vascular smooth muscle cells by induction of RNA synthesis. AB - This study examines the involvement of RNA and protein synthesis in the modulation of apoptosis in vascular smooth muscle cells (VSMC) by intracellular monovalent cations. In VSMC transfected with E1A adenovirus (VSMC-E1A), inversion of the [Na(+)](i)/[K(+)](i) ratio by an inhibitor of the Na(+),K(+) pump, ouabain, prevented the development of apoptosis triggered by serum withdrawal. Inhibition of apoptosis by ouabain was abolished by inhibitors of RNA and protein synthesis, actinomycin D, and cycloheximide, respectively. In VSMC-E1A, incubation with ouabain for 4 and 24 hours augmented RNA synthesis by 20% to 50% and 3-fold to 4-fold, respectively. In quiescent VSMC, the effect of ouabain and serum on RNA synthesis was additive. Ouabain did not affect the level of phosphorylation of ERK, JNK, and p38 MAP kinases and blocked apoptosis independent of the presence of the MAPK kinase inhibitors PD98059 and SB 202190. Equimolar substitution of NaCl with KCl in the incubation medium abolished the effect of ouabain on intracellular Na(+) and K(+) concentration, apoptosis, and RNA synthesis. Thus, our results demonstrate that the antiapoptotic effect of the inverted [Na(+)](i)/[K(+)](i) ratio is mediated by MAPK-independent induction of de novo synthesis of RNA species encoding inhibitor(s) of programmed cell death. PMID- 10818066 TI - Proapoptotic and growth-inhibitory role of angiotensin II type 2 receptor in vascular smooth muscle cells of spontaneously hypertensive rats in vivo. AB - Angiotensin type 2 (AT(2)) receptors for angiotensin II suppress cell growth and induce apoptosis in vitro, but their role is poorly defined in vivo. We reported that transient induction of smooth muscle cell (SMC) apoptosis precedes DNA synthesis inhibition and aortic hypertrophy regression in spontaneously hypertensive rats treated with the AT(1) antagonist losartan or the converting enzyme inhibitor enalapril. Although both drugs are equipotent in reducing SMC number, apoptosis occurs significantly earlier with losartan than enalapril. To examine the role of AT(2) receptors in this model, spontaneously hypertensive rats were given valsartan, an AT(1) antagonist, or enalapril, in combination or not with the AT(2) antagonist PD123319 for 1 or 2 weeks. Control rats received vehicle. Systolic blood pressure was reduced similarly by valsartan and enalapril but it was not significantly affected by PD123319. Angiotensin II plasma levels were increased (6-fold) with valsartan and reduced (80%) with enalapril but unaffected by PD123319. Valsartan significantly increased internucleosomal DNA fragmentation indicative of apoptosis at 1 week only (2.7-fold) and significantly reduced aortic mass (18%), SMC number (33%), and DNA synthesis (24%, measured by (3)H-thymidine incorporation) at 2 weeks. These valsartan-induced changes were prevented by PD123319. In contrast, enalapril-induced DNA fragmentation (2-fold increase at 2 weeks) was not affected by PD123319. PD123319 given alone did not affect growth or apoptosis. AT(1) and AT(2) receptor mRNAs were detected in the aorta by reverse transcription-polymerase chain reaction. Together, these results provide the first evidence that AT(2) receptors mediate vascular mass regression by stimulating SMC apoptosis in vivo, an effect seen during AT(1) receptor blockade but not during converting-enzyme inhibition. PMID- 10818067 TI - Angiotensin type 2 receptor mediates valsartan-induced hypotension in conscious rats. AB - Inhibition of the renin-angiotensin system is associated with vasodilation and reduction in blood pressure. We hypothesized that angiotensin type 1 (AT(1)) receptor (AT(1)R) blockade is associated with increased production of renal nitric oxide (NO) mediated by release of bradykinin (BK). By use of a microdialysis technique, changes in renal interstitial fluid (RIF) BK, NO end products nitrite and nitrate (NOX), and cGMP were monitored in response to intravenous infusion of the AT(1)R blocker valsartan (10 mg/kg), the angiotensin type 2 (AT(2)) receptor (AT(2)R) blocker PD123319 (50 microg x kg(-1) x min(-1)), and the BK B(2) receptor blocker icatibant (10 microg x kg(-1) x min(-1)) in conscious rats (n=10) during low sodium intake. RIF BK, NOX, and cGMP significantly increased during valsartan treatment, whereas AT(2)R blockade caused a significant decrease in these autacoids. During icatibant infusion, RIF NOX and cGMP decreased by 64% and 40%, respectively, whereas BK increased. Combined administration of valsartan and icatibant, of valsartan and PD123319, or of valsartan, PD123319, and icatibant prevented the increase in RIF cGMP and NOX in response to valsartan alone. These data demonstrate that AT(1)R blockade with valsartan is associated with release of renal BK, which in turn mediates NO production. The results suggest that increased angiotensin II, in response to sodium restriction and valsartan infusion, stimulates AT(2)R, which mediates a BK and NO cascade. PMID- 10818068 TI - Local angiotensin II and transforming growth factor-beta1 in renal fibrosis of rats. AB - Studies have demonstrated that local angiotensin II (Ang II) generation is enhanced in repairing kidney and that ACE inhibition or AT(1) receptor blockade attenuates renal fibrosis. The localization of ACE and Ang II receptors and their relationship to collagen synthesis in the injured kidney, however, remain uncertain. Using a rat model of renal injury with subsequent fibrosis created with chronic elevations in circulating aldosterone (ALDO), we examined the distribution and binding density of ACE and Ang II receptors in repairing kidneys, as well as their anatomic relationship to transforming growth factor beta1 (TGF-beta1) mRNA, type I collagen mRNA, collagen accumulation, and myofibroblasts. Two groups of animals (n=7 in each group) were studied: (1) normal rats served as controls, and (2) uninephrectomized rats received ALDO (0.75 microg/h SC) and 1% NaCl in drinking water for 6 weeks. Compared with control rats, in ALDO-treated rats we found (1) significantly (P<0.01) increased blood pressure, reduced plasma renin activity, and increased plasma creatinine levels, (2) diffuse fibrosis in both renal cortex and medulla, (3) abundant myofibroblasts at these sites of fibrosis, (4) significantly increased (P<0.01) binding density of ACE and Ang II receptors (60% AT(1), 40% AT(2)) at the sites of fibrosis, and (5) markedly increased (P<0.01) expression of TGF-beta1 and type I collagen mRNAs at these same sites. Thus, in this rat model of renal repair, the enhanced expression of ACE, Ang II receptors, and TGF-beta1 is associated with renal fibrosis. Ang II generated at the sites of repair appears to have autocrine/paracrine functions in the regulation of renal fibrous tissue formation alone or through its stimulation of TGF-beta1 synthesis. PMID- 10818069 TI - Effects of aspirin-like drugs on nitric oxide synthesis in rat vascular smooth muscle cells. AB - The purpose of this study was to investigate the effects of aspirin-like drugs on nitric oxide (NO) synthesis in rat vascular smooth muscle cells (VSMCs). We measured the accumulation of nitrite, a stable oxidation product of NO, and the expression of inducible NO synthase (iNOS) mRNA and protein in rat cultured VSMCs. Sodium salicylate, aspirin, and indomethacin dose-dependently enhanced nitrite production by interleukin (IL)-1beta-stimulated VSMCs at therapeutic plasma concentration ranges. Increased nitrite production by aspirin-like drugs was accompanied by increased iNOS mRNA and protein accumulation in VSMCs. Addition of IL-1beta activated nuclear factor kappaB (NF-kappaB) in VSMCs, but sodium salicylate did not affect IL-1beta-induced NF-kappaB activation. The nonselective lipoxygenase (LO) inhibitor nordihydroguaiaretic acid inhibited sodium salicylate-induced nitrite production, whereas the selective 5-LO inhibitor caffeic acid did not influence production of nitrite. The 12-LO product 12-HETE dose-dependently enhanced nitrite production by IL-1beta-stimulated VSMCs, whereas the 15-LO product 15-HETE did not. Our study demonstrates that aspirin and the aspirin-like drugs, sodium salicylate and indomethacin, increase NO synthesis in IL-1beta-stimulated VSMCs by upregulation of iNOS transcription via a 12-LO pathway. These effects were independent of NF-kappaB activation. In addition to the direct inhibition of platelet function, aspirin-like drugs may contribute to the reduction of atherothrombotic risk in myocardial ischemia via enhancing NO production by VSMCs. PMID- 10818070 TI - Aldose reductase inhibitor improves insulin-mediated glucose uptake and prevents migration of human coronary artery smooth muscle cells induced by high glucose. AB - We examined involvement of the polyol pathway in high glucose-induced human coronary artery smooth muscle cell (SMC) migration using Boyden's chamber method. Chronic glucose treatment for 72 hours potentiated, in a concentration-dependent manner (5.6 to 22.2 mol/L), platelet-derived growth factor (PDGF) BB-mediated SMC migration. This potentiation was accompanied by an increase in PDGF BB binding, because of an increased number of PDGF-beta receptors, and this potentiation was blocked by the aldose reductase inhibitor epalrestat. Epalrestat at concentrations of 10 and 100 nmol/L inhibited high glucose-potentiated (22.2 mmol/L), PDGF BB-mediated migration. Epalrestat at 100 nmol/L inhibited a high glucose-induced increase in the reduced/oxidized nicotinamide adenine dinucleotide ratio and membrane-bound protein kinase C (PKC) activity in SMCs. PKC inhibitors calphostin C (100 nmol/L) and chelerythrine (1 micromol/L) each inhibited high glucose-induced, PDGF BB-mediated SMC migration. High glucose induced suppression of insulin-mediated [(3)H]-deoxyglucose uptake, which was blocked by both calphostin C (100 nmol/L) and chelerythrine (1 micromol/L), was decreased by epalrestat (100 nmol/L). Chronic high glucose treatment for 72 hours increased intracellular oxidative stress, which was directly measured by flow cytometry using carboxydichlorofluorescein diacetate bis-acetoxymethyl ester, and this increase was significantly suppressed by epalrestat (100 nmol/L). Antisense oligonucleotide to PKC-beta isoform inhibited high glucose-mediated changes in SMC migration, insulin-mediated [(3)H]-deoxyglucose uptake, and oxidative stress. These findings suggest that high glucose concentrations potentiate SMC migration in coronary artery and that the aldose reductase inhibitor epalrestat inhibits high glucose-potentiated, PDGF BB-induced SMC migration, possibly through suppression of PKC (PKC-beta), impaired insulin-mediated glucose uptake, and oxidative stress. PMID- 10818071 TI - Role of 11beta-hydroxysteroid dehydrogenase in nongenomic aldosterone effects in human arteries. AB - The aim of the present study was to demonstrate rapid effects of aldosterone on the Na(+)-H(+) exchanger in strips of human vascular vessels and to determine whether 11beta-hydroxysteroid dehydrogenase enzyme (11beta-HSD) could play a protective role in this response, such as that described for the classic type I mineralocorticoid receptor (MR). The activity of 11beta-HSD isoforms 1 and 2 were measured in fetal and adult arteries. Both isoforms are present in adult and fetal vessels. However, a significant difference in the proportion of each isoform was found. Isoform 1 activity (in pmol x min(-1) x 100 mg(-1) protein) was 42+/-5 in fetal vessels and 29+/-2 in adult arteries, and isoform 2 activity was 78+/-7 in fetal and 12+/-2 in adult tissue. The nongenomic effect of aldosterone on Na(+)-H(+) exchanger activity was measured in strips of chorionic and radial uterine arteries loaded with the pH-sensitive dye 2',7'-bis(2 carboxyethyl)-5,6-carboxyfluorescein. Recordings of intracellular pH (pH(i)) were made by videofluorescence microscopy. Aldosterone (0.5 nmol/L) rapidly increased pH(i), with a half-maximal effect between 2 and 3 nmol/L in both fetal and adult vessels. Ethylisopropylamiloride, a specific inhibitor of the Na(+)-H(+) exchanger, inhibited this effect. The hormone-mediated increase in pH(i) was unaffected by spironolactone, a classic antagonist of MR, but was completely blocked by RU28318. Cortisol (up to 1 micromol/L) had no effect on pH(i), but when applied in the presence of carbenoxolone, a dramatic increase in Na(+)-H(+) exchanger activity was evident. The increments on pH(i) for each cortisol concentration were similar to those observed for aldosterone. These findings suggest that vascular 11beta-HSD plays an active role in maintaining the specificity of the rapid effects of aldosterone. PMID- 10818072 TI - Effect of lovastatin on cerebral circulation in spontaneously hypertensive rats. AB - Statins, which are often given to hypertensive patients, reduce the incidence of stroke. However, their effects on the cerebral circulation have been scarcely studied, although lovastatin has been reported to reduce hypertension-induced renal arteriolar hypertrophy. We examined the structure and mechanics of cerebral arterioles and the lower limit of cerebral blood flow (CBF) autoregulation in spontaneously hypertensive rats (SHR) that were untreated (n=9) or treated for 1 month with lovastatin (n=12; 20 mg x kg(-1) x d(-1)) and in untreated Wistar Kyoto rats (WKY; n=8). We studied the lower limit of CBF autoregulation by repeated measurement of CBF (arbitrary units; laser Doppler) and internal arteriolar diameter (microm; cranial window) at baseline and during stepwise hypotension. Stress-strain relationships were calculated from repeated measurement of internal arteriolar diameter during stepwise hypotension and cross sectional area (CSA) of the vessel wall in maximally dilated cerebral arterioles (EDTA, 67 mmol/L). Lovastatin slightly reduced mean arterial pressure (treated, 153+/-3 versus untreated, 171+/-5 mm Hg, P<0.05; WKY, 106+/-3 mm Hg) and normalized CSA (treated, 826+/-52 versus untreated, 1099+/-16 microm(2), P<0. 05; WKY, 774+/-28 microm(2)). Stress-strain curves show that lovastatin also attenuated the increase in passive distensibility. Lovastatin had no effect on the external diameter of cerebral arterioles or the lower limit of CBF autoregulation. Our results show that although lovastatin has substantial effects on arteriolar mechanics and wall CSA, it has little effect on internal diameter. This phenomenon may explain its lack of effect on CBF autoregulation. PMID- 10818073 TI - Nitric oxide deficiency contributes to large cerebral infarct size. AB - The purpose of this study was to examine the role played by a deficit in nitric oxide (NO) in contributing to the large cerebral infarcts seen in hypertension. Cerebral infarction was produced in rats by occlusion of the middle cerebral artery (MCA). Studies were performed in Sprague-Dawley (SD) rats subjected to NO synthase blockade (N(G)-nitro-L-arginine [L-NNA], 20 mg x kg(-1) x d(-1) in drinking water) and in spontaneously hypertensive stroke-prone rats (SHRSP). NO released in the brain in response to MCA occlusion was monitored with a porphyrinic microsensor in Wistar-Kyoto rats. The increment in NO released with MCA occlusion was 1.31+/-0.05 micromol/L in L-NNA-treated rats, 1.25+/-0.04 micromol/L in SHRSP, 2. 24+/-0.07 micromol/L in control SD rats, and 2.25+/-0.06 micromol/L in Wistar-Kyoto rats (P<0.0001 for control versus the other groups). Infarct sizes in the L-NNA-treated and control SD rats were 8.50+/-0. 8% and 5.22+/-0.7% of the brain weights, respectively (P<0.05). The basilar arterial wall was significantly thicker in L-NNA-treated rats compared with their controls. We conclude that both the deficit in NO and the greater wall thickness contribute to the larger infarct size resulting from MCA occlusion in SHRSP and in L-NNA-treated rats compared with their respective controls. PMID- 10818074 TI - Atrial natriuretic peptide modifies arterial blood pressure through nitric oxide pathway in rats. AB - The aim of the present study was to determine the relationship between the hypotensive effect of the atrial natriuretic peptide (ANP) and the nitric oxide (NO) pathway. N(G)-nitro-L-arginine methyl ester bolus (L-NAME, 1 mg/kg) reverted the decrease in mean arterial pressure induced by ANP administration (5 microg/kg bolus and 0.2 microg x kg(-1) x min(-1) infusion), and the injection of L-NAME before peptide administration suppressed the ANP hypotensive response. To confirm these findings, a histochemical reaction was used to determine NADPH-diaphorase activity (a NO synthase marker) in the endothelium and smooth muscle of aorta and arterioles of the small and large intestine. ANP increased aorta and arteriole endothelium staining after both in vivo administration and in vitro tissue incubation. In both cases, L-NAME prevented the ANP effect on NADPH-diaphorase activity. Tissues incubated with 8-bromoguanosine 3',5'-cyclic monophosphate mimicked ANP action. In addition, ANP administration increased urinary excretion of NO(x) end products. These findings indicate that ANP increases NO synthesis capability and NO production and suggest that the cGMP pathway may be involved. In conclusion, the NO pathway could be an intercellular messenger in the ANP endothelium-dependent vasorelaxation mechanism. PMID- 10818075 TI - Estimation of skeletal muscle interstitial adenosine during forearm dynamic exercise in humans. AB - It has been proposed that adenosine is a metabolic signal that triggers activation of muscle afferents involved in the exercise pressor reflex. Furthermore, exogenous adenosine induces sympathetic activation that mimics the exercise pressor reflex, and blockade of adenosine receptors inhibits sympathetic activation induced by exercise. Thus, we hypothesize that adenosine is released locally by the muscle during exercise. We used microdialysis probes, placed in the flexor digitorium superficialis muscle, to estimate muscle interstitial adenosine levels in humans. We estimated resting in vivo muscle interstitial adenosine concentrations (0.292+/-0.058 micromol/L, n=4) by perfusing increasing concentrations of adenosine to determine the gradient produced in the dialysate. Muscle interstitial adenosine concentrations increased from 0.23+/-0.04 to 0.82+/ 0.14 micromol/L (n=14, P<0.001) during intermittent dynamic exercise at 50% of maximal voluntary contraction. Lactate increased from 0.8+/-0.1 to 2.3+/-0.3 mmol/L (P<0.001). Lower intensity (15% maximal voluntary contraction) intermittent dynamic exercise increased adenosine concentrations from 0.104+/ 0.02 to 0.42+/-0.16 micromol/L (n=7). The addition of ischemia to this low level of exercise produced a greater increase in adenosine (from 0.095+/-0.02 to 0.48+/ 0.2 micromol/L) compared with nonischemic exercise (0. 095+/-0.02 to 0.25+/-0.12 micromol/L). These results indicate that microdialysis is useful in estimating adenosine concentrations and in reflecting changes in muscle interstitial adenosine during dynamic exercise in humans. PMID- 10818076 TI - Characterization of murine vasopressor and vasodepressor prostaglandin E(2) receptors. AB - Four E-prostanoid (EP) receptors, designated EP(1), EP(2), EP(3), and EP(4), mediate the cellular effects of prostaglandin E(2) (PGE(2)). The present studies pharmacologically characterize the vasopressor and vasodepressor EP receptors in wild-type mice (EP(2)(+/+) mice) and mice with targeted disruption of the EP(2) receptor (EP(2)(-/-) mice). Mean arterial pressure (MAP) was measured via a carotid artery catheter in anesthetized male mice. Intravenous infusion of PGE(2) decreased MAP in EP(2)(+/+) mice but increased MAP in EP(2)(-/-) mice. Infusion of EP(3)-selective agonists, including MB28767, SC46275, and sulprostone, increased MAP in both EP(2)(+/+) and EP(2)(-/-) mice. Pretreatment with SC46275 desensitized mice to the subsequent pressor effect of sulprostone, but the vasodepressor effect of PGE(2) in EP(2)(+/+) mice remained intact. Although PGE(2) alone increased MAP in EP(2)(-/-) mice, prior desensitization of the pressor effect with SC46275 allowed a residual vasodepressor effect of PGE(2) to be seen in the EP(2)(-/-) mice. An EP(4)-selective agonist (prostaglandin E(1) OH) functioned also as a vasodepressor in both EP(2)(-/-) and EP(2)(+/+) mice. High levels of EP(3) receptor mRNA were detected in mouse aortas and rabbit preglomerular arterioles by nuclease protection, with lower expressions of EP(1), EP(2), and EP(4) mRNA. The findings suggest that combined vasodepressor effects of EP(2) and EP(4) receptors normally dominate, accounting for the depressor effects of PGE(2). In contrast, in EP(2)(-/-) mice, EP(4) receptor activity alone is insufficient to overcome the EP(3) vasopressor effect. These findings suggest that a balance between pressor and depressor PGE(2) receptors determines its net effect on arterial pressure and that these receptors may be important therapeutic targets. PMID- 10818077 TI - Weight gain-induced blood pressure elevation. AB - This study was conducted to evaluate the mechanisms in weight gain-induced blood pressure (BP) elevation focusing, in particular, on the contributions of sympathetic nervous system activity, fasting plasma insulin, and leptin to BP levels. The study design was longitudinal with a cohort of 1897 men. BP, pulse rate, body mass index (BMI), fasting plasma norepinephrine (NE), insulin, and leptin were measured at 6 and 12 months in those 172 lean normotensive, 79 obese normotensive, 64 lean untreated hypertensive, and 38 obese untreated hypertensive men whose BMI increased >10% during the first 6 months. At entry, levels of BP, pulse rate, plasma NE, insulin, and leptin in obese subjects, regardless of BP status, were significantly greater than those in lean subjects. The levels of plasma NE, insulin, and leptin increased with weight gain in the 4 study groups. In the subjects with BP elevation, the increase in pulse rate and plasma NE was significantly greater than that in the subjects without BP elevation at both 6 and 12 months for each of the 4 study groups, although the increase in BMI was similar between the subjects with and without BP elevation. In obese but not lean subjects, whether normotensive or hypertensive, the increases in plasma insulin and plasma leptin with weight gain were greater in the subjects with accompanying BP elevation compared with the subjects without BP elevation. On the other hand, at 6 months in lean subjects, the increase in plasma insulin with weight gain in the subjects with BP elevation was actually lower than that in the subjects without BP elevation. These results suggest that weight gain-induced sympathetic overactivity is more tightly linked to weight gain-induced BP elevation than the changes in plasma insulin and leptin that also accompany weight gain. It is probable that sympathetic nervous activation with weight gain is a major mechanism of blood pressure elevation. Hyperinsulinemia and hyperleptinemia may be ancillary factors that contribute to sympathetic nervous stimulation with weight gain. PMID- 10818078 TI - Heritability of angiotensin-converting enzyme and angiotensinogen: A comparison of US blacks and Nigerians. AB - Angiotensinogen (AGT) and angiotensin I-converting enzyme (ACE) are heritable traits, but whether the environmental context influences heritability has not been examined. Known genetic factors explain only a portion of variation in AGT and ACE, and levels of both proteins are influenced by the environment. The African diaspora provides an opportunity to compare these traits in genetically related populations in contrasting environments. As part of a study of the genetics of hypertension, we examined families that included 1449 Nigerians and 1147 African Americans. Body mass index (weight [kg]/height [m](2)) was 21 kg/m(2) in Nigeria and 29 kg/m(2) in the United States, which is consistent with a large environmental contrast. AGT was considerably higher among African Americans (1919 versus 1396, P<0.01), whereas ACE was higher in Nigerians (630 versus 517, P<0.01). A household effect was observed among the Nigerian families (spouse correlations 0.30 for AGT, 0.18 for ACE), and correlations among first degree relatives were large (0.42 to 0. 51 and 0.36 to 0.38 for AGT and ACE, respectively). Among African Americans, the familial aggregations of AGT and ACE were very limited, and the familial correlation for AGT was not different from zero. Heritability was 77% for AGT and 67% for ACE in Nigeria and 18% for AGT and ACE in the United States. The familial patterns of body mass index and blood pressure were similar among both family sets. In conclusion, less familial aggregation was observed for AGT and ACE in the United States than in Nigeria, most likely reflecting a greater random individual environmental effect on these traits. Variation in heritability of traits could influence the power of epidemiological studies to identify genetic effects. PMID- 10818079 TI - Antihypertensive effects of fasidotril, a dual inhibitor of neprilysin and angiotensin-converting enzyme, in rats and humans. AB - The aim of this study was to assess the antihypertensive activity of fasidotril, a dual inhibitor of neprilysin (NEP) and angiotensin I-converting enzyme (ACE), in various models of hypertension in rats (spontaneously hypertensive rats [SHR]; renovascular Goldblatt 2-kidney, 1-clip rats; and deoxycorticosterone acetate [DOCA]-salt hypertensive rats) and in patients with mild-to-moderate essential hypertension. Fasidotril treatment (100 mg/kg PO twice daily for 3 weeks) resulted in a progressive and sustained decrease in systolic blood pressure (-20 to -30 mm Hg) in SHR and Goldblatt rats compared with vehicle-treated rats and prevented the progressive rise in blood pressure in DOCA-salt hypertensive rats. After a 4-week placebo run-in period, 57 patients with essential hypertension were included in a randomized double-blind, placebo-controlled, parallel-group study and received orally either fasidotril (100 mg twice daily) or placebo for 6 weeks. Blood pressure was measured during the 6 hours after the first intake and then at trough (12 hours after the last intake) on days 7, 28, and 42. The first dose of fasidotril had no significant effect on blood pressure. After 42 days, compared with placebo, fasidotril lowered supine systolic and diastolic blood pressures by 7.4/5.4 mm Hg and standing blood pressure by 7.6/6.8 mm Hg. Fasidotril, a dual NEP/ACE inhibitor, was an effective oral antihypertensive agent during chronic treatment in high-renin renovascular rats, normal-renin SHR, and low-renin DOCA-salt hypertensive rats and in patients with essential hypertension. PMID- 10818080 TI - Relation between low calcium intake, parathyroid hormone, and blood pressure. AB - In a population health survey in 1995, serum parathyroid hormone (PTH) was measured in 1113 subjects, aged 30 to 79 years, and was found to be elevated (>6.9 pmol/L) in 118 subjects. In 1998, this group and 131 subjects with normal PTH levels were invited for reexamination, and 82 and 90 subjects from each respective group attended the follow-up. At the follow-up, 72 subjects had elevated and 100 had normal serum PTH levels. Those with elevated serum PTH levels (8 subjects with hyperparathyroidism were excluded) had significantly lower serum calcium levels and intake of calcium than those with normal PTH (2.24+/-0.09 and 2.29+/-0.10 mmol/L [mean+/-SD] and 400.3+/-227.3 and 592.1+/ 459.6 mg/d, respectively; P<0.01). Serum levels or intake of vitamin D did not differ between the 2 groups. Subjects with elevated PTH in both 1995 and 1998 had significantly lower bone mineral content and bone mineral density in the lumbar spine than did those with persistently normal PTH levels (P<0.05). In the females, but not in the males, the systolic and diastolic blood pressures were significantly higher in those with elevated serum PTH (158.0+/-27.5 versus 141.5+/-19.2 mm Hg and 90. 5+/-13.6 versus 82.6+/-8.6 mm Hg, respectively; P<0.01). This difference was even more pronounced when those with persistently elevated PTH were considered separately. In conclusion, reduced intake of calcium is frequently associated with high levels of serum PTH. This is associated with moderately reduced bone mineral content and bone mineral density in the lumbar spine. In women, high levels of serum PTH are also associated with markedly increased blood pressure. PMID- 10818081 TI - Activation of MAPKs in proximal tubule cells from spontaneously hypertensive and control Wistar-Kyoto rats. AB - The aim of this study was to test the hypothesis that differences exist in the activity and/or expression of mitogen-activated protein kinases (MAPKs) between spontaneously hypertensive rats (SHR) and control Wistar-Kyoto rats (WKY) and that these differences may account for the enhanced activity of the Na(+)/H(+) exchanger (NHE) previously observed in the renal proximal tubule of SHR. Therefore, the activities of c-jun N-terminal kinase(1) (JNK(1)), extracellular signal-regulated kinase(1/2) (ERK(1/2)), and p38 were investigated. A reduced amount of ERK(1) and JNK(1) protein was found in renal cortex specimens of SHR as compared with WKY; however, their activities were the same. To study the cellular basis of this difference, immortalized proximal tubule cell lines were grown on Millicell-CM filter inserts where the cell lines organize as polarized monolayers with separate access to apical and basolateral compartments. Although basal JNK(1) and ERK(1/2) activities were not significantly different between WKY and SHR cells, anisomycin stimulated JNK(1) activity in WKY cells more than in SHR cells (eg, at 15 minutes 300% versus 30%, respectively). Similarly, angiotensin II increased JNK(1) and ERK(1/2) activity in a time- and concentration-dependent manner in WKY cells but not in SHR cells. Western blot analyses showed a deficit in JNK(1) and ERK(1) protein in SHR (0.25 and 0.5, respectively, of the levels in WKY cells), although ERK(2) and p38 protein levels were the same. These observations suggest that, although angiotensin II activates MAPKs and MAPKs have been shown to regulate NHE, this regulatory pathway is unlikely to account for the increased activity of NHE in the proximal tubular epithelium of SHR. PMID- 10818082 TI - Comparison of hearts with 2 types of pressure-overload left ventricular hypertrophy. AB - Comparisons of myocardium remodeled by the 2 most common causes of left ventricular hypertrophy (LVH), hypertension and aortic constriction, are limited. We hypothesized that important differences may exist in the myocardium of hearts with these 2 origins of "pressure overload" LVH. Accordingly, we studied isolated hearts from 3 groups of Dahl salt-sensitive rats, controls, and hearts with matched amounts of LVH secondary to either hypertension or aortic constriction. Isovolumic LV function and myocardial energetics ((31)P nuclear magnetic resonance spectroscopy) were measured as coronary flow was lowered to 16% of baseline for 48 minutes. During this low-flow ischemia, isovolumic end-diastolic pressure, a measure of LV stiffness, increased to 52+/-4 mm Hg in controls and 51+/-6 mm Hg in aortic banded hearts but to only 35+/-5 mm Hg in hearts with hypertensive LVH. In all hearts, the P(i) resonance in the (31)P nuclear magnetic resonance spectrum, whose position indicates myocardial pH, split into 2 peaks during low-flow ischemia, which indicates distinct regions of pH 6.9 (moderate acidosis) and pH 6.2 (severe acidosis). Concentrations of ATP, PCr, P(i), and H(+) of the moderately acidotic region were not different among groups. However, the size of the severely acidotic region was smallest in the hypertensive LVH hearts, and in all 3 groups, the size of this region correlated (r(2)=0.65 to 0.80) with the degree of LV stiffening. We conclude that in Dahl rats, LVH secondary to hypertension protects against ischemia-induced diastolic dysfunction by minimizing the size of the region of severe acidosis. PMID- 10818084 TI - Hypertension online only : may 2000 PMID- 10818083 TI - Effects of sleep deprivation on neural circulatory control. AB - Effects of sleep deprivation on neural cardiovascular control may have important clinical implications. We tested the hypothesis that sleep deprivation increases heart rate, blood pressure, and sympathetic activity and potentiates their responses to stressful stimuli. We studied 8 healthy subjects (aged 40+/-5 years, 6 men and 2 women). Blood pressure, heart rate, forearm vascular resistance, and muscle sympathetic nerve activity were measured at rest and during 4 stressors (sustained handgrip, maximal forearm ischemia, mental stress, and cold pressor test). Measurements were obtained twice, once after normal sleep and once after a night of sleep deprivation. All measurements were obtained in a blinded, randomized manner. In comparison with normal sleep, sleep deprivation resulted in an increase in blood pressure (normal sleep versus sleep deprivation=82+/-8 versus 86+/-7 mm Hg, mean+/-SEM, P=0.012) and a decrease in muscle sympathetic nerve activity (normal sleep versus sleep deprivation=28+/-6 versus 22+/-6 bursts/min, P=0.017). Heart rate, forearm vascular resistance, and plasma catecholamines were not significantly changed by sleep deprivation, nor did sleep deprivation affect autonomic and hemodynamic responses to stressful stimuli. Sleep deprivation results in increased resting blood pressure, decreased muscle sympathetic nerve activity, and no change in heart rate. Thus, the pressor response to sleep deprivation is not mediated by muscle sympathetic vasoconstriction or tachycardia. PMID- 10818085 TI - J-shaped relation between blood pressure and stroke. PMID- 10818086 TI - Mitochondrial translocation of protein kinase C delta in phorbol ester-induced cytochrome c release and apoptosis. AB - Apoptosis is induced by the release of cytochrome c from mitochondria to the cytoplasm. The present studies demonstrate that the phorbol ester 12-O tetradecanoylphorbol-13-acetate (TPA) induces translocation of protein kinase C (PKC) delta from the cytoplasm to mitochondria. The results also show that translocation of PKCdelta results in release of cytochrome c. The functional significance of this event is further supported by the demonstration that PKCdelta translocation is required for TPA-induced apoptosis. These findings demonstrate that translocation of PKCdelta to mitochondria is responsible, at least in part, for inducing cytochrome c release and apoptosis. PMID- 10818087 TI - Single amino acid substitutions in kappa-conotoxin PVIIA disrupt interaction with the shaker K+ channel. AB - kappa-Conotoxin PVIIA (kappa-PVIIA), a 27-amino acid peptide with three disulfide cross-links, isolated from the venom of Conus purpurascens, is the first conopeptide shown to inhibit the Shaker K(+) channel (Terlau, H., Shon, K., Grilley, M., Stocker, M., Stuhmer, W., and Olivera, B. M. (1996) Nature 381, 148 151). Recently, two groups independently determined the solution structure for kappa-PVIIA using NMR; although the structures reported were similar, two mutually exclusive models for the interaction of the peptide with the Shaker channel were proposed. We carried out a structure/function analysis of kappa PVIIA, with alanine substitutions for all amino acids postulated to be key residues by both groups. Our data are consistent with the critical dyad model developed by Menez and co-workers (Dauplais, M., Lecoq, A., Song, J. , Cotton, J., Jamin, N., Gilquin, B., Roumestand, C., Vita, C., de Medeiros, C., Rowan, E. G., Harvey, A. L., and Menez, A. (1997) J. Biol. Chem. 272, 4802-4809) for polypeptide antagonists of K(+) channels. In the case of kappa-PVIIA, Lys(7) and Phe(9) are essential for activity as predicted by Savarin et al. (Savarin, P., Guenneugues, M., Gilquin, B., Lamthanh, H., Gasparini, S., Zinn-Justin, S., and Menez, A. (1998) Biochemistry 37, 5407-5416); these workers also correctly predicted an important role for Lys(25). Thus, although kappa-conotoxin PVIIA has no obvious sequence homology to polypeptide toxins from other venomous animals that interact with voltage-gated K(+) channels, there may be convergent functional features in diverse K(+) channel polypeptide antagonists. PMID- 10818088 TI - The homeodomain coordinates nuclear entry of the Lhx3 neuroendocrine transcription factor and association with the nuclear matrix. AB - LIM homeodomain transcription factors regulate development in complex organisms. To characterize the molecular signals required for the nuclear localization of these proteins, we examined the Lhx3 factor. Lhx3 is essential for pituitary organogenesis and motor neuron specification. By using functional fluorescent derivatives, we demonstrate that Lhx3 is found in both the nucleoplasm and nuclear matrix. Three nuclear localization signals were mapped within the homeodomain, and one was located in the carboxyl terminus. The homeodomain also serves as the nuclear matrix targeting sequence. No individual signal is alone required for nuclear localization of Lhx3; the signals work in combinatorial fashion. Specific combinations of these signals transferred nuclear localization to cytoplasmic proteins. Mutation of nuclear localization signals within the homeodomain inhibited Lhx3 transcriptional function. By contrast, mutation of the carboxyl-terminal signal activated Lhx3, indicating that this region is critical to transcriptional activity and may be a target of regulatory pathways. The pattern of conservation of the nuclear localization and nuclear matrix targeting signals suggests that the LIM homeodomain factors use similar mechanisms for subcellular localization. Furthermore, upon nuclear entry, association of Lhx3 with the nuclear matrix may contribute to LIM homeodomain factor interaction with other classes of transcription factors. PMID- 10818089 TI - A CCAAT/enhancer-binding protein site within antioxidant/electrophile response element along with CREB-binding protein participate in the negative regulation of rat GST-Ya gene in vascular smooth muscle cells. AB - Studies were conducted to evaluate the negative regulatory function of rat (r)GST Ya antioxidant/electrophile response element (ARE/EpRE) in vascular smooth muscle cells (vSMCs). We report that CCAAT/enhancer-binding protein (C/EBP)-beta interacts with ARE/EpRE in the rGST-Ya promoter and that aryl hydrocarbon receptor (AhR) is present within the protein complex binding to the C/EBP site. Overexpression of C/EBP-beta or C/EBP-alpha repressed, whereas AhR enhanced, 1.6CAT reporter activity in cells treated with benzo(a)pyrene (BaP). Overexpression of CREB-binding protein (CBP) nullified repression of rGST-Ya transcription. Human adenovirus E1A protein abrogated cotransactivation by CBP but an E1A mutant did not. Overexpression of C/EBPs abrogated stimulation of 1.6CAT by CBP or AhR alone, or in combination, regardless of BaP treatment. Similar profiles were observed using an AhRECAT construct. The C/EBP site within the ARE/EpRE inhibited chemical inducibility of the AhRE. The pattern of mouse GST-Ya regulation by BaP was similar to that of rGST-Ya. We conclude that multiple mechanisms mediate negative regulation of GST-Ya gene in vSMCs, most significant of which are that C/EBP-beta inhibits AhRE or ARE/EpRE inducibility of GST-Ya, limiting CBP levels compromise gene induction, functional interference exists between AhRE and ARE/EpRE, and AhR alone, or in combination with C/EBP beta, functions as a repressor of the ARE/EpRE. PMID- 10818090 TI - The PSI-K subunit of photosystem I is involved in the interaction between light harvesting complex I and the photosystem I reaction center core. AB - PSI-K is a subunit of photosystem I. The function of PSI-K was characterized in Arabidopsis plants transformed with a psaK cDNA in antisense orientation, and several lines without detectable PSI-K protein were identified. Plants without PSI-K have a 19% higher chlorophyll a/b ratio and 19% more P700 than wild-type plants. Thus, plants without PSI-K compensate by making more photosystem I. The photosystem I electron transport in vitro is unaffected in the absence of PSI-K. Light response curves for oxygen evolution indicated that the photosynthetic machinery of PSI-K-deficient plants have less capacity to utilize light energy. Plants without PSI-K have less state 1-state 2 transition. Thus, the redistribution of absorbed excitation energy between the two photosystems is reduced. Low temperature fluorescence emission spectra revealed a 2-nm blue shift in the long wavelength emission in plants lacking PSI-K. Furthermore, thylakoids and isolated PSI without PSI-K had 20-30% less Lhca2 and 30-40% less Lhca3, whereas Lhca1 and Lhca4 were unaffected. During electrophoresis under mildly denaturing conditions, all four Lhca subunits were partially dissociated from photosystem I lacking PSI-K. The observed effects demonstrate that PSI-K has a role in organizing the peripheral light-harvesting complexes on the core antenna of photosystem I. PMID- 10818091 TI - Tau phosphorylation at serine 396 and serine 404 by human recombinant tau protein kinase II inhibits tau's ability to promote microtubule assembly. AB - In Alzheimer's disease, hyperphosphorylated tau is an integral part of the neurofibrillary tangles that form within neuronal cell bodies and fails to promote microtubule assembly. Dysregulation of the brain-specific tau protein kinase II is reported to play an important role in the pathogenesis of Alzheimer's disease (Patrick, G. N., Zukerberg, L., Nikolic, M., De La Monte, S., Dikkes, P., and Tsai, L.-H. (1999) Nature 402, 615-622). We report here that in vitro phosphorylation of human tau by human recombinant tau protein kinase II severely inhibits the ability of tau to promote microtubule assembly as monitored by tubulin polymerization. The ultrastructure of tau-mediated polymerized tubulin was visualized by electron microscopy and compared with phosphorylated tau. Consistent with the observed slower kinetics of tubulin polymerization, phosphorylated tau is compromised in its ability to generate microtubules. Moreover, we show that phosphorylation of microtubule-associated tau results in tau's dissociation from the microtubules and tubulin depolymerization. Mutational studies with human tau indicate that phosphorylation by tau protein kinase II at serine 396 and serine 404 is primarily responsible for the functional loss of tau mediated tubulin polymerization. These in vitro results suggest a possible role for tau protein kinase II-mediated tau phosphorylation in initiating the destabilization of microtubules. PMID- 10818092 TI - Targeting the ets binding site of the HER2/neu promoter with pyrrole-imidazole polyamides. AB - Three DNA binding polyamides () were synthesized that bind with high affinity (K(a) = 8.7. 10(9) m(-1) to 1.4. 10(10) m(-1)) to two 7-base pair sequences overlapping the Ets DNA binding site (EBS; GAGGAA) within the regulatory region of the HER2/neu proximal promoter. As measured by electrophoretic mobility shift assay, polyamides binding to flanking elements upstream () or downstream (2 and 3) of the EBS were one to two orders of magnitude more effective than the natural product distamycin at inhibiting formation of complexes between the purified EBS protein, epithelial restricted with serine box (ESX), and the HER2/neu promoter probe. One polyamide, 2, completely blocked Ets-DNA complex formation at 10 nm ligand concentration, whereas formation of activator protein-2-DNA complexes was unaffected at the activator protein-2 binding site immediately upstream of the HER2/neu EBS, even at 100 nm ligand concentration. At equilibrium, polyamide 1 was equally effective at inhibiting Ets/DNA binding when added before or after in vitro formation of protein-promoter complexes, demonstrating its utility to disrupt endogenous Ets-mediated HER2/neu preinitiation complexes. Polyamide 2, the most potent inhibitor of Ets-DNA complex formation by electrophoretic mobility shift assay, was also the most effective inhibitor of HER2/neu promoter driven transcription measured in a cell-free system using nuclear extract from an ESX- and HER2/neu-overexpressing human breast cancer cell line, SKBR-3. PMID- 10818093 TI - Phosphorylation of collapsin response mediator protein-2 by Rho-kinase. Evidence for two separate signaling pathways for growth cone collapse. AB - We previously identified Rho-associated protein kinase (Rho-kinase) as a specific effector of Rho. In this study, we identified collapsin response mediator protein 2 (CRMP-2), as a novel Rho-kinase substrate in the brain. CRMP-2 is a neuronal protein whose expression is up-regulated during development. Rho-kinase phosphorylated CRMP-2 at Thr-555 in vitro. We produced an antibody that specifically recognizes CRMP-2 phosphorylated at Thr-555. Using this antibody, we found that Rho-kinase phosphorylated CRMP-2 downstream of Rho in COS7 cells. Phosphorylation of CRMP-2 was observed in chick dorsal root ganglion neurons during lysophosphatidic acid (LPA)-induced growth cone collapse, whereas the phosphorylation was not detected during semaphorin-3A-induced growth cone collapse. Both LPA-induced CRMP-2 phosphorylation and LPA-induced growth cone collapse were inhibited by Rho-kinase inhibitor HA1077 or Y-32885. LPA-induced growth cone collapse was also blocked by a dominant negative form of Rho-kinase. On the other hand, semaphorin-3A-induced growth cone collapse was not inhibited by a dominant negative form of Rho-kinase. Furthermore, overexpression of a mutant CRMP-2 in which Thr-555 was replaced by Ala significantly inhibited LPA induced growth cone collapse. These results demonstrate the existence of Rho kinase-dependent and -independent pathways for growth cone collapse and suggest that CRMP-2 phosphorylation by Rho-kinase is involved in the former pathway. PMID- 10818094 TI - Evidence for gelsolin as a corneal crystallin in zebrafish. AB - We have shown that gelsolin is one of the most prevalent water-soluble proteins in the transparent cornea of zebrafish. There are also significant amounts of actin. In contrast to actin, gelsolin is barely detectable in other eye tissues (iris, lens, and remaining eye) of the zebrafish. Gelsolin cDNA hybridized intensely in Northern blots to RNA from the cornea but not from the lens, brain, or headless body. The deduced zebrafish gelsolin is approximately 60% identical to mammalian cytosolic gelsolin and has the characteristic six segmental repeats as well as the binding sites for actin, calcium, and phosphatidylinositides. In situ hybridization tests showed that gelsolin mRNA is concentrated in the zebrafish corneal epithelium. The zebrafish corneal epithelium stains very weakly with rhodamine-phalloidin, indicating little F-actin in the cytoplasm. In contrast, the mouse corneal epithelium contains relatively little gelsolin and stains intensely with rhodamine-phalloidin, as does the zebrafish extraocular muscle. We propose, by analogy with the diverse crystallins of the eye lens and with the putative enzyme-crystallins (aldehyde dehydrogenase class 3 and other enzymes) of the mammalian cornea, that gelsolin and actin-gelsolin complexes act as water-soluble crystallins in the zebrafish cornea and contribute to its optical properties. PMID- 10818095 TI - The J-helix of Escherichia coli DNA polymerase I (Klenow fragment) regulates polymerase and 3'- 5'-exonuclease functions. AB - To assess the functional importance of the J-helix region of Escherichia coli DNA polymerase I, we performed site-directed mutagenesis of the following five residues: Asn-675, Gln-677, Asn-678, Ile-679, and Pro-680. Of these, the Q677A mutant is polymerase-defective with no change in its exonuclease activity. In contrast, the N678A mutant has unchanged polymerase activity but shows increased mismatch-directed exonuclease activity. Interestingly, mutation of Pro-680 has a Q677A-like effect on polymerase activity and an N678A-like effect on the exonuclease activity. Mutation of Pro-680 to Gly or Gln results in a 10-30-fold reduction in k(cat) on homo- and heteropolymeric template-primers, with no significant change in relative DNA binding affinity or K(m)((dNTP)). The mutants P680G and P680Q also showed a nearly complete loss in the processive mode of DNA synthesis. Since the side chain of proline is generally non-reactive, mutation of Pro-680 may be expected to alter the physical form of the J-helix itself. The biochemical properties of P680G/P680Q together with the structural observation that J-helix assumes helical or coiled secondary structure in the polymerase or exonuclease mode-bound DNA complexes suggest that the structural alteration in the J-helix region may be responsible for the controlled shuttling of DNA between the polymerase and the exonuclease sites. PMID- 10818096 TI - Autoinhibition of a calmodulin-dependent calcium pump involves a structure in the stalk that connects the transmembrane domain to the ATPase catalytic domain. AB - The regulation of Ca(2+)-pumps is important for controlling [Ca(2+)] in the cytosol and organelles of all eukaryotes. Here, we report a genetic strategy to identify residues that function in autoinhibition of a novel calmodulin-activated Ca(2+)-pump with an N-terminal regulatory domain (isoform ACA2 from Arabidopsis). Mutant pumps with constitutive activity were identified by complementation of a yeast (K616) deficient in two Ca(2+)-pumps. Fifteen mutations were found that disrupted a segment of the N-terminal autoinhibitor located between Lys(23) and Arg(54). Three mutations (E167K, D219N, and E341K) were found associated with the stalk that connects the ATPase catalytic domain (head) and with the transmembrane domain. Enzyme assays indicated that the stalk mutations resulted in calmodulin independent activity, with V(max), K(mATP), and K(mCa(2+)) similar to that of a pump in which the N-terminal autoinhibitor had been deleted. A highly conservative substitution at Asp(219) (D219E) still produced a deregulated pump, indicating that the autoinhibitory structure in the stalk is highly sensitive to perturbation. In plasma membrane H(+)-ATPases from yeast and plants, similarly positioned mutations resulted in hyperactive pumps. Together, these results suggest that a structural feature of the stalk is of general importance in regulating diverse P-type ATPases. PMID- 10818097 TI - Identities and phylogenetic comparisons of posttranscriptional modifications in 16 S ribosomal RNA from Haloferax volcanii. AB - Small subunit (16 S) rRNA from the archaeon Haloferax volcanii, for which sites of modification were previously reported, was examined using mass spectrometry. A census of all modified residues was taken by liquid chromatography/electrospray ionization-mass spectrometry analysis of a total nucleoside digest of the rRNA. Following rRNA hydrolysis by RNase T(1), accurate molecular mass values of oligonucleotide products were measured using liquid chromatography/electrospray ionization-mass spectrometry and compared with values predicted from the corresponding gene sequence. Three modified nucleosides, distributed over four conserved sites in the decoding region of the molecule, were characterized: 3-(3 amino-3-carboxypropyl)uridine-966, N(6)-methyladenosine-1501, and N(6),N(6) dimethyladenosine-1518 and -1519 (all Escherichia coli numbering). Nucleoside 3 (3-amino-3-carboxypropyl)uridine, previously unknown in rRNA, occurs at a highly conserved site of modification in all three evolutionary domains but for which no structural assignment in archaea has been previously reported. Nucleoside N(6) methyladenosine, not previously placed in archaeal rRNAs, frequently occurs at the analogous location in eukaryotic small subunit rRNA but not in bacteria. H. volcanii small subunit rRNA appears to reflect the phenotypically low modification level in the Crenarchaeota kingdom and is the only cytoplasmic small subunit rRNA shown to lack pseudouridine. PMID- 10818098 TI - Wild-type but not Parkinson's disease-related ala-53 --> Thr mutant alpha synuclein protects neuronal cells from apoptotic stimuli. AB - Recent works suggest that alpha-synuclein could play a central role in Parkinson's disease (PD). Thus, two mutations were reported to be associated with rare autosomal dominant forms of the disease. We examined whether alpha-synuclein could modulate the caspase-mediated response and vulnerability of murine neurons in response to various apoptotic stimuli. We established TSM1 neuronal cell lines overexpressing wild-type (wt) alpha-synuclein or the PD-related Ala-53 --> Thr mutant alpha-synuclein. Under basal conditions, acetyl-Asp-Glu-Val-Asp-aldehyde sensitive caspase activity appears significantly lower in wt alpha-synuclein expressing cells than in neurons expressing the mutant. Interestingly, wt alpha synuclein drastically reduces the caspase activation of TSM1 neurons upon three distinct apoptotic stimuli including staurosporine, etoposide, and ceramide C(2) when compared with mock-transfected cells. This inhibitory control of the caspase response triggered by apoptotic agents was abolished by the PD-related pathogenic mutation. Comparison of wild-type and mutated alpha-synuclein-expressing cells also indicates that the former exhibits much less vulnerability in response to staurosporine and etoposide as measured by the sodium 3'-[1-(phenylaminocarbonyl) 3, 4-tetrazolium]-bis(4-methoxy-6-nitro)benzenesulfonic acid assay. Altogether, our study indicates that wild-type alpha-synuclein exerts an antiapoptotic effect in neurons that appears to be abolished by the Parkinson's disease-related mutation, thereby suggesting a possible mechanism underlying both sporadic and familial forms of this neurodegenerative disease. PMID- 10818099 TI - A short C-terminal domain of Yku70p is essential for telomere maintenance. AB - The Yku heterodimer from Saccharomyces cerevisiae, comprising Yku70p and Yku80p, is involved in the maintenance of a normal telomeric DNA end structure and is an essential component of nonhomologous end joining (NHEJ). To investigate the role of the Yku70p subunit in these two different pathways, we generated C-terminal deletions of the Yku70 protein and examined their ability to complement the phenotypes of a yku70(-) strain. Deleting only the 30 C-terminal amino acids of Yku70p abolishes Yku DNA binding activity and causes a yku(-) phenotype; telomeres are shortened, and NHEJ is impaired. Using conditions in which at least as much mutant protein as full-length protein is normally detectable in cell extracts, deleting only 25 C-terminal amino acids of Yku70p results in no measurable effect on DNA binding of the Yku protein, and the cells are fully proficient for NHEJ. Nevertheless, these cells display considerably shortened telomeres, and significant amounts of single-stranded overhangs of the telomeric guanosine-rich strands are observed. Co-overexpression of this protein with Yku80p could rescue some but not all of the telomere-related phenotypes. Therefore, the C-terminal domain in Yku70p defines at least one domain that is especially involved in telomere maintenance but not in NHEJ. PMID- 10818100 TI - Endoplasmic reticulum oxidoreductin 1-lbeta (ERO1-Lbeta), a human gene induced in the course of the unfolded protein response. AB - Oxidative conditions must be generated in the endoplasmic reticulum (ER) to allow disulfide bond formation in secretory proteins. A family of conserved genes, termed ERO for ER oxidoreductins, plays a key role in this process. We have previously described the human gene ERO1-L, which complements several phenotypic traits of the yeast thermo-sensitive mutant ero1-1 (Cabibbo, A., Pagani, M., Fabbri, M., Rocchi, M., Farmery, M. R., Bulleid, N. J., and Sitia, R. (2000) J. Biol. Chem. 275, 4827-4833). Here, we report the cloning and characterization of a novel human member of this family, ERO1-Lbeta. Immunofluorescence, endoglycosidase sensitivity, and in vitro translation/translocation assays reveal that the products of the ERO1-Lbeta gene are primarily localized in the ER of mammalian cells. The ability to allow growth at 37 degrees C and to alleviate the "unfolded protein response" when expressed in ero1-1 cells indicates that ERO1 Lbeta is involved also in generating oxidative conditions in the ER. ERO1-L and ERO1-Lbeta display different tissue distributions. Furthermore, only ERO1-Lbeta transcripts are induced in the course of the unfolded protein response. Our results suggest a complex regulation of ER redox homeostasis in mammalian cells. PMID- 10818101 TI - Molecular recognition of taxol by microtubules. Kinetics and thermodynamics of binding of fluorescent taxol derivatives to an exposed site. AB - We have determined the kinetic scheme and the reaction rates of binding to microtubules of two fluorescent taxoids, 7-O-[N-(4'-fluoresceincarbonyl)-l alanyl]Taxol (Flutax-1) and 7-O-[N-(2,7-difluoro-4'-fluoresceincarbonyl)-l alanyl]Taxol (Flutax-2). Flutax-1 and Flutax-2 bind to microtubules with high affinity (K(a) approximately 10(7) m(-1), 37 degrees C). The binding mechanism consists of a fast bimolecular reaction followed by at least two monomolecular rearrangements, which were characterized with stopped-flow techniques. The kinetic constants of the bimolecular reaction were 6.10 +/- 0.22 x 10(5) m(-1) s( 1) and 13.8 +/- 1.8 x 10(5) m(-1) s(-1) at 37 degrees C, respectively. A second slow binding step has been measured employing the change of fluorescence anisotropy of the probe. The reversal of this reaction is the rate-limiting step of dissociation. A third step has been detected using small angle x-ray scattering and involves a 2-nm increase in the diameter of microtubules. It is suggested that the first step entails the binding of the Taxol moiety and the second a relative immobilization of the fluorescent probe. The equilibrium and some kinetic measurements required the use of stabilized cross-linked microtubules, which preserved taxoid binding. The results indicate that the Taxol binding site is directly accessible, in contrast with its location at lumen in the current model of microtubules. An alternative structural model is considered in which the binding site is located between protofilaments, accessible from the microtubule surface. PMID- 10818102 TI - MEK kinase 2 binds and activates protein kinase C-related kinase 2. Bifurcation of kinase regulatory pathways at the level of an MAPK kinase kinase. AB - MEK kinase 2 (MEKK2) is a 70-kDa protein serine/threonine kinase that has been shown to function as a mitogen-activated protein kinase (MAPK) kinase kinase. MEKK2 has its kinase domain in the COOH-terminal moiety of the protein. The NH(2) terminal moiety of MEKK2 has no signature motif that would suggest a defined regulatory function. Yeast two-hybrid screening was performed to identify proteins that bind MEKK2. Protein kinase C-related kinase 2 (PRK2) was found to bind MEKK2; PRK2 has been previously shown to bind RhoA and the Src homology 3 domain of Nck. PRK2 did not bind MEKK3, which is closely related to MEKK2. The MEKK2 binding site maps to amino acids 637-660 in PRK2, which is distinct from the binding sites for RhoA and Nck. This sequence is divergent in the closely related kinase PRK1, which did not bind MEKK2. In cells, MEKK2 and PRK2 are co immunoprecipitated and PRK2 is activated by MEKK2. Similarly, purified recombinant MEKK2 activated PRK2 in vitro. MEKK2 activation of PRK2 is independent of MEKK2 regulation of the c-Jun NH(2)-terminal kinase pathway. MEKK2 activation of PRK2 results in a bifurcation of signaling for the dual control of MAPK pathways and PRK2 regulated responses. PMID- 10818103 TI - Evolutionarily conserved features of the arginine attenuator peptide provide the necessary requirements for its function in translational regulation. AB - Neurospora crassa arg-2 mRNA contains an evolutionarily conserved upstream open reading frame (uORF) encoding the Arg attenuator peptide (AAP) that confers negative translational regulation in response to Arg. We examined the regulatory role of the AAP and the RNA encoding it using an N. crassa cell-free translation system. AAPs encoded by uORFs in four fungal mRNAs each conferred negative regulation in response to Arg by causing ribosome stalling at the uORF termination codon. Deleting the AAP non-conserved N terminus did not impair regulation, but deletions extending into the conserved region eliminated it. Introducing many silent mutations into a functional AAP coding region did not eliminate regulation, but a single additional nucleotide change altering the conserved AAP sequence abolished regulation. Therefore, the conserved peptide sequence, but not the mRNA sequence, appeared responsible for regulation. AAP extension at its C terminus resulted in Arg-mediated ribosomal stalling during translational elongation within the extended region and during termination. Comparison of Arg-mediated stalling at a rare or common codon revealed more stalling at the rare codon. These data indicate that the highly evolutionarily conserved peptide core functions within the ribosome to cause stalling; translational events at a potential stall site can influence the extent of stalling there. PMID- 10818104 TI - Identification and molecular cloning of a chondroitin synthase from Pasteurella multocida type F. AB - Pasteurella multocida Type F, the minor fowl cholera pathogen, produces an extracellular polysaccharide capsule that is a putative virulence factor. It was reported that the capsule was removed by treating microbes with chondroitin AC lyase. We found by acid hydrolysis that the polysaccharide contained galactosamine and glucuronic acid. We molecularly cloned a Type F polysaccharide synthase and characterized its enzymatic activity. The 965-residue enzyme, called P. multocida chondroitin synthase (pmCS), is 87% identical at the nucleotide and the amino acid level to the hyaluronan synthase, pmHAS, from P. multocida Type A. A recombinant Escherichia coli-derived truncated, soluble version of pmCS (residues 1-704) was shown to catalyze the repetitive addition of sugars from UDP GalNAc and UDP-GlcUA to chondroitin oligosaccharide acceptors in vitro. Other structurally related sugar nucleotide precursors did not substitute in the elongation reaction. Polymer molecules composed of approximately 10(3) sugar residues were produced, as measured by gel filtration chromatography. The polysaccharide synthesized in vitro was sensitive to the action of chondroitin AC lyase but resistant to the action of hyaluronan lyase. This is the first report identifying a glycosyltransferase that forms a polysaccharide composed of chondroitin disaccharide repeats, [beta(1,4)GlcUA-beta(1,3)GalNAc](n). In analogy to known hyaluronan synthases, a single polypeptide species, pmCS, possesses both transferase activities. PMID- 10818105 TI - Regulation of galpha i palmitoylation by activation of the 5-hydroxytryptamine-1A receptor. AB - Nearly all alpha subunits of heterotrimeric GTP-binding regulatory proteins (G proteins) are palmitoylated at cysteine residues near the N terminus. A regulated cycle of palmitoylation could provide a mechanism for modulating G protein signaling by affecting protein interactions and localization of the subunit. In the present studies we utilized both [(3)H]palmitate incorporation and pulse chase techniques to address the dynamics of alpha(i) palmitoylation in Chinese hamster ovary cells. Both techniques demonstrated a dose- and time-dependent change in [(3)H]palmitate labeling of alpha(i) upon activation of stably expressed 5-hydroxytryptamine-1A receptors by the agonist (+/-)-2-dipropylamino-8 hydroxy-1,2,3, 4-tetrahydronaphthalene hydrobromide (DPAT), with an EC(50) of approximately 10 nm. For the incorporation assay, DPAT elicited an approximate doubling in labeling at the earliest time point measured. For the pulse-chase assay, DPAT promoted a significant loss of radiolabel almost equally as fast. These data demonstrate that the exchange of palmitate on alpha(i) is increased upon stimulation of 5-hydroxytryptamine-1A receptors through the combined processes of depalmitoylation and palmitoylation. These results provide the basis for extending the concept of regulated exchange of palmitate beyond G(s) and provide a framework for exploring the specific functional attributes of the palmitoylated and depalmitoylated forms of subunit. PMID- 10818106 TI - Identification of a novel sequence involved in lysosomal sorting of the sphingolipid activator protein prosaposin. AB - Prosaposin is synthesized as a 53-kDa protein, post-translationally modified to a 65-kDa form and further glycosylated to a 70-kDa secretory product. The 65-kDa protein is associated to Golgi membranes and is targeted to lysosomes, where four smaller nonenzymatic saposins implicated in the hydrolysis of sphingolipids are generated by its partial proteolysis. The targeting of the 65-kDa protein to lysosomes is not mediated by the mannose 6-phosphate receptor. The Golgi apparatus appears to accomplish the molecular sorting of the 65-kDa prosaposin by decoding a signal from its amino acid backbone. This investigation deals with the characterization of the sequence involved in this process by deleting the saposin functional domains A, B, C, and D and the highly conserved N and C termini of prosaposin. The truncated cDNAs were subcloned into expression vectors and transfected to COS-7 cells. The destination of the mutated proteins was assessed by immunocytochemistry. Deletion of the C terminus did not interfere with the secretion of prosaposin but abolished its transport to lysosomes. Deletion of saposins and the N-terminal domain did not affect the lysosomal or secretory routing of prosaposin. A chimeric construct of albumin and the C terminus of prosaposin was not directed to lysosomes. However, albumin connected to the C terminus and one or more functional domains of prosaposin reached lysosomes, indicating that the C terminus and at least one saposin domain are required for this process. In summary, we are reporting a novel sequence involved in the targeting of prosaposin to lysosomes. PMID- 10818107 TI - A Di-leucine signal in the ubiquitin moiety. Possible involvement in ubiquitination-mediated endocytosis. AB - Some plasma membrane receptors in yeast are known to be internalized and degraded in lysosomes upon ligand-dependent ubiquitination. However, the role of ubiquitination in endocytosis and lysosomal degradation in higher eukaryotes has been controversial. In order to directly assess this question, we investigated the fate of chimeric molecules in which ubiquitin moiety was fused in-frame to the cytoplasmic region of membrane proteins. The chimeric proteins with the wild type ubiquitin were endocytosed and delivered to lysosomes efficiently. Mutant ubiquitin with lysine-to-arginine substitution could still mediate endocytosis, suggesting that polyubiquitination is not required for the endocytosis. We next searched for the existence of an endocytosis signal(s) in the ubiquitin moiety, and identified a di-leucine signal, Leu(43)-Ile(44). The Leu(43)-Ile(44) sequence mediated endocytosis and lysosomal sorting in a Leu(43)-dependent manner. These results suggest that the di-leucine signal in ubiquitin can be involved in ubiquitination-mediated endocytosis and lysosomal targeting of membrane proteins. PMID- 10818108 TI - Localization of the cyclic ADP-ribose-dependent calcium signaling pathway in hepatocyte nucleus. AB - CD38 is a type II transmembrane glycoprotein found on both hematopoietic and non hematopoietic cells. It is known for its involvement in the metabolism of cyclic ADP-ribose (cADPR) and nicotinic acid adenine dinucleotide phosphate, two nucleotides with calcium mobilizing activity independent of inositol trisphosphate. It is generally believed that CD38 is an integral protein with ectoenzymatic activities found mainly on the plasma membrane. Here we show that enzymatically active CD38 is present intracellularly on the nuclear envelope of rat hepatocytes. CD38 isolated from rat liver nuclei possessed both ADP-ribosyl cyclase and NADase activity. Immunofluorescence studies on rat liver cryosections and isolated nuclei localized CD38 to the nuclear envelope of hepatocytes. Subcellular localization via immunoelectron microscopy showed that CD38 is located on the inner nuclear envelope. The isolated nuclei sequestered calcium in an ATP-dependent manner. cADPR elicited a rapid calcium release from the loaded nuclei, which was independent of inositol trisphosphate and was inhibited by 8 amino-cADPR, a specific antagonist of cADPR, and ryanodine. However, nicotinic acid adenine dinucleotide phosphate failed to elicit any calcium release from the nuclear calcium stores. The nuclear localization of CD38 shown in this study suggests a novel role of CD38 in intracellular calcium signaling for non hematopoietic cells. PMID- 10818109 TI - Perlecan heparan sulfate proteoglycan: a novel receptor that mediates a distinct pathway for ligand catabolism. AB - Cell surface heparan sulfate proteoglycans (HSPGs) participate in the catabolism of many physiologically important ligands. We previously reported that syndecan HSPGs directly mediate endocytosis, independent of coated pits. We now studied perlecan, a major cell surface HSPG genetically distinct from syndecans. Cells expressing perlecan but no other proteoglycans bound, internalized, and degraded atherogenic lipoproteins enriched in lipoprotein lipase. Binding was blocked by heparitinase, and degradation by chloroquine. Antibodies against beta(1) integrins reduced initial ligand binding, consistent with their roles as cell surface attachment sites for perlecan. By several criteria, catabolism via perlecan was distinct from either coated pits or the syndecan pathway. The kinetics of internalization (t(12) = 6 h) and degradation (t(12) approximately 18 h) were remarkably slow, unlike the other pathways. Blockade of the low density lipoprotein receptor-related protein did not slow perlecan-dependent internalization. Internalization via perlecan was inhibited by genistein but unaffected by cytochalasin D, a pattern distinct from coated pits or syndecan mediated endocytosis. Finally, we examined cooperation between perlecan and low density lipoprotein receptors and found limited synergy. Our results demonstrate that perlecan mediates internalization and lysosomal delivery that is kinetically and biochemically distinct from other known uptake pathways and is consistent with a very slow component of HSPG-dependent ligand processing found in vitro and in vivo. PMID- 10818111 TI - Social capital: the role of narrative and historical research. PMID- 10818110 TI - A novel membrane-anchored Rab5 interacting protein required for homotypic endosome fusion. AB - The ras-related GTPase rab5 is rate-limiting for homotypic early endosome fusion. We used a yeast two-hybrid screen to identify a rab5 interacting protein, rab5ip. The cDNA sequence encodes a ubiquitous 75-kDa protein with an N-terminal transmembrane domain (TM), a central coiled-coil structure, and a C-terminal region homologous to several centrosome-associated proteins. rab5ip lacking the transmembrane domain (rab5ipTM(-)) had a greater affinity in vitro for rab5 guanosine 5'-O-2-(thio)diphosphate than for rab5-guanosine 5'-3-O (thio)triphosphate. In transfected HeLa cells, rab5ipTM(-) was partly cytosolic and localized (by immunofluorescence) with a rab5 mutant believed to be in a GDP conformation (GFP-rab5(G78A)) but not with GFP-rab5(Q79L), a GTPase-deficient mutant. rab5ip with the transmembrane domain (rab5ipTM(+)) was completely associated with the particulate fraction and localized extensively with GFP rab5(wt) in punctate endosome-like structures. Overexpression of rab5ipTM(+) using Sindbis virus stimulated the accumulation of fluid-phase horseradish peroxidase by BHK-21 cells, and homotypic endosome fusion in vitro was inhibited by antibody against rab5ip. rab5ipTM(-) inhibited rab5(wt)-stimulated endosome fusion but did not inhibit fusion stimulated by rab5(Q79L). rab5ip represents a novel rab5 interacting protein that may function on endocytic vesicles as a receptor for rab5-GDP and participate in the activation of rab5. PMID- 10818112 TI - Real or perceived adverse effects of vaccines and the media--a tale of our times. PMID- 10818113 TI - Social capital--is it a good investment strategy for public health? PMID- 10818114 TI - Social capital, economic capital and power: further issues for a public health agenda. PMID- 10818115 TI - Inequality and the social environment: a reply to Lynch et al. PMID- 10818116 TI - Epidemiology of participation: an Australian community study. AB - STUDY OBJECTIVE: To determine the levels of participation in social and civic community life in a metropolitan region, and to assess differential levels of participation according to demographic, socioeconomic and health status. To contribute to policy debates on community participation, social capital and health using these empirical data. DESIGN: Cross sectional, postal, self completed survey on health and participation. SETTING: Random sample of the population from the western suburbs of Adelaide, the capital city of South Australia, a population of approximately 210 000. PARTICIPANTS: 2542 respondents from a sample of 4000 people aged 18 years and over who were registered on the electoral roll. MAIN RESULTS: The response rate to the survey was 63.6% (n=2542). Six indices of participation, on range of social and civic activities, with a number of items in each, were created. Levels of participation were highest in the informal social activities index (46.7-83.7% for individual items), and lowest in the index of civic activities of a collective nature (2.4-5.9% for individual items). Low levels of involvement in social and civic activities were reported more frequently by people of low income and low education levels. CONCLUSIONS: Levels of participation in social and civic community life in an urban setting are significantly influenced by individual socioeconomic status, health and other demographic characteristics. An understanding of the pattern of participation is important to inform social and health policy making. Increasing levels of participation will reduce social exclusion and is likely to improve the overall quality of community life. PMID- 10818117 TI - Obesity and other health determinants across Europe: the EURALIM project. AB - STUDY OBJECTIVE: EURALIM (EURope ALIMentation), a European collaborative study, aimed to determine and describe the extent to which European data on risk factor distributions from different populations could be pooled and harmonised in a common database for international comparisons. SETTING: Seven independent population-based surveys from six European countries (France, Italy, Northern Ireland/United Kingdom, Spain, Switzerland, the Netherlands). METHODS: Data for 18 381 women and 12 908 men aged 40-59 were pooled in a common database. Central statistical analyses on major cardiovascular risk factors were conducted with careful consideration of methodological issues, including differences in study designs, data assessment tools, and analytic techniques used. MAIN RESULTS: Because of the detected variability among methods used, direct comparisons of risk factor distributions and prevalences between studies were problematic. None the less, comparisons of within population contrasts by sex, age group, and other health determinants were considered to be meaningful and apt, as illustrated here for obesity. Results were targeted and disseminated to both the general public and public health professionals and framed in the context of a European information campaign. CONCLUSIONS: International and national comparisons between existing locally run studies are feasible and useful, but harmonisation methods need improvement. Development of an international risk factor surveillance programme based on decentralised data collection is warranted. In the meantime, risk factor contrasts across populations can be used as a basis for targeting needed public health intervention programmes. PMID- 10818118 TI - Haematopoietic cancer and medical history: a multicentre case control study. AB - BACKGROUND: Viruses (such as Epstein-Barr virus) and pathological conditions (mainly involving immunosuppression) have been shown to increase the risk of haematolymphopoietic malignancies. Other associations (diabetes, tonsillectomy, autoimmune diseases) have been inconsistently reported. METHODS: The association between different haematolymphopoietic malignancies (lymphomas, myelomas and leukaemias) and the previous medical history has been studied in a population based case-control investigation conducted in Italy, based on face to face interviews to 2669 cases and 1718 population controls (refusal rates 10% and 19%, respectively). Controls were a random sample of the general population. RESULTS: Previous findings were confirmed concerning the association between non-Hodgkin's lymphoma (NHL) and lupus erythematosus (odds ratio, OR=8.4; 95% CI 1. 6, 45), tuberculosis (OR=1.6; 1.05, 2.5) and hepatitis (1.8; 1.4, 2. 3). An association was found also between NHL and maternal (OR=2.8; 1.1, 6.9) or paternal tuberculosis (OR=1.7; 0.7, 3.9). Odds ratios of 4.0 (1.4, 11.8) and 4.4 (1.1, 6.6) were detected for the association between NHL and Hodgkin's disease, respectively, and previous infectious mononucleosis, but recall bias cannot be ruled out. No association was found with diabetes, tonsillectomy and adenoidectomy. An association with malaria at young age and "low grade" lymphatic malignancies is suggested. One interesting finding was the observation of four cases of poliomyelitis among NHL patients, one among Hodgkin's disease and one among myeloid leukaemia patients, compared with none among the controls (Fisher's exact test for NHL and Hodgkin's disease, p= 0.03, one tail). CONCLUSIONS: Some of these findings are confirmatory of previous evidence. Other observations, such as the putative role of the polio virus and of malaria are new. A unifying theory on the mechanisms by which previous medical history may increase the risk of haematolymphopoietic malignancies is still lacking. PMID- 10818119 TI - Changes in social inequalities in health in the Basque Country. AB - STUDY OBJECTIVE: To determine the extent of the inequalities in self reported health between socioeconomic groups and its changes over time in the Basque Country (Spain). DESIGN: Cross sectional data on the association between occupation, education and income and three health indicators was obtained from the Basque Health Surveys of 1986 and 1992. Representative population samples were analysed. In 1986 the number of respondents was 24 657 and in 1992, 13 277. SETTING: Basque Country, Spain. MAIN OUTCOME MEASURES: The effect of socioeconomic position on health and the magnitude of social inequalities in health were quantified using the odds ratios based on logistic regression analysis, and the Relative Index of Inequality. RESULTS: As was expected, social inequalities in self reported health existed in both surveys, but the social gradient was greater in 1992. Social differences varied according to gender and health indicator. According to education an increase in social inequalities was observed consistently in all the health indicators except long term conditions in women. A consistent increase in inequalities in limiting longstanding illness was also observed according to all socioeconomic indicators. CONCLUSIONS: These results agree to a large extent with those of previous studies in other countries. In this context the unequal distribution of material circumstances and working conditions between socioeconomic groups seem to play a major part in health inequalities. The worsening of the labour market during this period and the onset of a new economic recession may explain the increase in social inequalities over time. PMID- 10818120 TI - Seroconversion and seroreversion in IgG antibodies to Helicobacter pylori: a serology based prospective cohort study. AB - STUDY OBJECTIVES: To assess the incidence of seroconversion and seroreversion in IgG antibodies to Helicobacter pylori within a 11 year observation period using these events as markers for acquisition and loss of the infection, respectively. DESIGN: Population based prospective cohort study. SETTING: Northern part of Copenhagen County, Denmark. PARTICIPANTS AND METHODS: A random sample of 2527 Danish adults were examined and blood obtained in 1983 and in 1994. Matching pairs of sera were analysed for the presence of IgG and IgM antibodies to H pylori with an in house enzyme linked immunosorbent assay. Participants who were seronegative at study entry and seropositive at follow up and had a fourfold increase in baseline IgG antibody levels were categorised as seroconverters and regarded as having acquired H pylori infection. Participants who were seropositive at study entry and had at least a fourfold decrease in baseline IgG antibody levels at follow up were assumed to have lost the infection (seroreverters). RESULTS: The seroprevalence of H pylori infection was 24.7 (95% confidence intervals (95% CI) 23.0, 26.4) % in 1983 and 24.5 (95% CI 22.8, 26. 2) % in 1994. A total of 14 participants seroconverted within the observation period (cumulative 11 year incidence proportion: 1.0 (95% CI 0.5, 1.5) %). Having increased IgM antibody levels at study entry significantly increased the likelihood of IgG seroconversion (relative risk 6.4 (95% CI 2.1, 19.6). Seroreversion was seen in 48 participants (cumulative 11 year incidence proportion: 7.7 (95% CI 5. 6, 9.8) %). CONCLUSIONS: Changes in H pylori infection status with time are rare in Danish adults. Few adults become infected with H pylori in Denmark. PMID- 10818121 TI - Factors explaining the geographical differences in Disability Free Life Expectancy in Spain. AB - OBJECTIVE: To study the geographical variations in Disability Free Life Expectancy (DFLE) at birth (DFLEb) and at 65 years (DFLE65) in Spain and to identify the main factors that explain these variations. DESIGN: Ecological study with the 50 provinces of Spain as the units of analysis. Sullivan's method is used to calculate DFLE for each province based on information from the death registry and the survey on disabilities, impairments and handicaps. Information on the independent variables-socioeconomic level, factors related with the health system and risk factors-was taken from various sources. MAIN OUTCOME MEASURE: Simple correlation coefficients were obtained between each dependent variable (DFLEb and DLFE65) and the independent variables. Two multiple linear regression models were fit to obtain the best set of factors that explain the geographical distribution of DFLEb and DLFE65. RESULTS: Both DFLEb and DLFE65 vary widely among provinces. The multiple linear regression analysis shows that the illiteracy rate, the percentage of the unemployed and the percentage of smokers in the population were the main factors associated with the geographical variation of DFLE. The models explained approximately 40% of the variance for DFLEb and 30% for DLFE65. CONCLUSIONS: The results obtained show the influence of education, the unemployment rate and smoking on the geographical differences of DFLE. The DFLE indicators are shown to be valid for use in health policy. PMID- 10818122 TI - Deprivation and childhood obesity: a cross sectional study of 20,973 children in Plymouth, United Kingdom. AB - OBJECTIVE: To study the association between socioeconomic deprivation and childhood obesity. DESIGN: Cross sectional study. SETTING: All state primary schools in Plymouth. Plymouth is a relatively deprived city in the United Kingdom, ranking 338th of 366 local authorities on the Department of the Environment Index of Local Conditions. SUBJECTS: 20 973 children between the ages of 5 and 14 years, 1994-96. MAIN OUTCOME MEASURE: Numbers of obese children (body mass index (BMI) above the 98th centile) by quarters of Townsend score. RESULTS: Plymouth had a rate of childhood obesity two and half times that expected nationally (5% v 2%). The obesity prevalence increased with age, being almost double in the oldest age quarter (boys 6.2%; girls 7.0%), compared with the youngest age quarter. Within Plymouth, there was a significant trend for higher rates of obesity related to increasing deprivation in both boys (p=0. 017) and girls (p=0.018). The odds ratio (OR) for childhood obesity (highest-lowest quarter of Townsend scores) had borderline significance in boys (OR 1.29, 95% confidence intervals (CI) 1.00 to 1.65, p=0.049) but was larger and more significant in the girls (OR 1.39, 95% CI 1.08 to 1.80, p=0.011). Unlike boys, the association between obesity in girls and Townsend scores became stronger with age such that in the oldest age quarter (over 11.7 years), girls in the highest quarter of Townsend scores were nearly twice as likely be obese, as compared with the lowest quarter (OR 1.95, 95% CI 1.23 to 3.08, p=0.005). State of pubertal development could not be accounted for as this information was not available. CONCLUSIONS: This study provides evidence for an association between deprivation and childhood obesity in this English population. The health of children from deprived households is affected by a number of adverse influences. The high prevalence of obesity in these children is yet another factor that could predispose to greater morbidity in adult life. PMID- 10818123 TI - How are policy makers using evidence? Models of research utilisation and local NHS policy making. AB - STUDY OBJECTIVE: This paper is based on a qualitative study that aimed to identify factors that facilitate or impede evidence-based policy making at a local level in the UK National Health Service (NHS). It considers how models of research utilisation drawn from the social sciences map onto empirical evidence from this study. DESIGN: A literature review and case studies of social research projects that were initiated by NHS health authority managers or GP fundholders in one region of the NHS. In depth interviews and document analysis were used. SETTING: One NHS region in England. PARTICIPANTS: Policy makers, GPs and researchers working on each of the social research projects selected as case studies. MAIN RESULTS: The direct influence of research evidence on decision making was tempered by factors such as financial constraints, shifting timescales and decision makers' own experiential knowledge. Research was more likely to impact on policy in indirect ways, including shaping policy debate and mediating dialogue between service providers and users. CONCLUSIONS: The study highlights the role of sustained dialogue between researchers and the users of research in improving the utilisation of research-based evidence in the policy process. PMID- 10818124 TI - Balkans briefing number 5. Seeking refuge (Macedonia, 9-25 June 1999): the changing needs of humanitarian aid in the face of the peace. PMID- 10818125 TI - Impact of a local newspaper campaign on the uptake of the measles mumps and rubella vaccine. PMID- 10818126 TI - Severity of fall injuries on sand or grass in playgrounds. PMID- 10818127 TI - Health care provision for illegal immigrants: should public health be concerned? PMID- 10818128 TI - The plasmin system is induced by and degrades amyloid-beta aggregates. AB - Amyloid-beta (Abeta) appears critical to Alzheimer's disease. To clarify possible mechanisms of Abeta action, we have quantified Abeta-induced gene expression in vitro by using Abeta-treated primary cortical neuronal cultures and in vivo by using mice transgenic for the Abeta precursor (AbetaP). Here, we report that aggregated, but not nonaggregated, Abeta increases the level of the mRNAs encoding tissue plasminogen activator (tPA) and urokinase-type plasminogen activator (uPA). Moreover, tPA and uPA were also upregulated in aged AbetaP overexpressing mice. Because others have reported that Abeta aggregates can substitute for fibrin aggregates in activating tPA post-translationally, the result of tPA induction by Abeta would be cleavage of plasminogen to the active protease plasmin. To gain insights into the possible actions of plasmin, we evaluated the hypotheses that tPA and plasmin may mediate Abeta in vitro toxicity or, alternatively, that plasmin activation may lead to Abeta degradation. In evaluating these conflicting hypotheses, we found that purified plasmin degrades Abeta with physiologically relevant efficiency, i.e., approximately 1/10th the rate of plasmin on fibrin. Mass spectral analyses show that plasmin cleaves Abeta at multiple sites. Electron microscopy confirms indirect assays suggesting that plasmin degrades Abeta fibrils. Moreover, exogenously added plasmin blocks Abeta neurotoxicity. In summation, we interpret these results as consistent with the possibility that the plasmin pathway is induced by aggregated Abeta, which can lead to Abeta degradation and inhibition of Abeta actions. PMID- 10818129 TI - Neuromodulatory effects of gonadotropin releasing hormone on olfactory receptor neurons. AB - The terminal nerve is an anterior cranial nerve that innervates the lamina propria of the chemosensory epithelia of the nasal cavity. The function of the terminal nerve is ambiguous, but it has been suggested to serve a neuromodulatory role. We tested this hypothesis by exposing olfactory receptor neurons from mudpuppies (Necturus maculosus) to a peptide, gonadotropin releasing hormone (GnRH), that is found in cells and fibers of the terminal nerve. We used voltage clamped whole-cell recordings to examine the effects of 0. 5-50 micrometer GnRH on voltage-activated currents in olfactory receptor neurons from epithelial slices. We found that GnRH increases the magnitude, but does not alter the kinetics, of a tetrodotoxin-sensitive inward current. This increase in magnitude generally begins 5-10 min after initial exposure to GnRH, is sustained for at least 60 min during GnRH exposure, and recovers to baseline within 5 min after GnRH is washed off. This effect occurred in almost 60% of the total number of olfactory receptor neurons examined and appeared to be seasonal: approximately 67% of neurons responded to GnRH during the courtship and mating season, compared with approximately 33% during the summer, when the sexes separate. GnRH also appears to alter an outward current in the same cells. Taken together, these data suggest that GnRH increases the excitability of olfactory receptor neurons and that the terminal nerve functions to modulate the odorant sensitivity of olfactory receptor neurons. PMID- 10818130 TI - Parallel cone bipolar pathways to a ganglion cell use different rates and amplitudes of quantal excitation. AB - The cone signal reaches the cat's On-beta (X) ganglion cell via several parallel circuits (bipolar cell types b1, b2, and b3). These circuits might convey different regions of the cone's temporal bandwidth. To test this, I presented a step of light that elicited a transient depolarization followed by a sustained depolarization. The contribution of bipolar cells to these response components was isolated by blocking action potentials with tetrodotoxin and by blocking inhibitory synaptic potentials with bicuculline and strychnine. Stationary fluctuation analysis of the sustained depolarization gave the rate of quantal bombardment: approximately 5100 quanta sec(-1) for small central cells and approximately 45,000 quanta sec(-1) for large peripheral cells. Normalizing these rates for the vastly different numbers of bipolar synapses (150-370 per small cell vs 2000 per large cell), quantal rate was constant across the retina, approximately 22 quanta synapse(-1) sec(-1). Nonstationary fluctuation analysis gave the mean quantal EPSP amplitude: approximately 240 microV for the transient depolarization and 30 microV for the sustained depolarization. The b1 bipolar cell is known from noise analysis of the On-alpha ganglion cell to have a near maximal sustained release of only approximately two quanta synapse(-1) sec(-1). This implies that the other bipolar types (b2 and b3) contribute many more quanta to the sustained depolarization (>/=46 synapse(-1) sec(-1)). Type b1 probably contributes large quanta to the transient depolarization. Thus, bipolar cell types b1 and b2/b3 apparently constitute parallel circuits that convey, respectively, high and low frequencies. PMID- 10818131 TI - Motoneuron activity patterns related to the earliest behavior of the zebrafish embryo. AB - As a first step in the study of the developing motor circuitry of the embryonic zebrafish spinal cord, we obtained patch-clamp recordings in vivo from identified motoneurons in curarized embryos from the onset of the first motor behavior. At an early developmental stage in which embryos showed slow and repetitive spontaneous contractions of the trunk, motoneurons showed periodic depolarizations that triggered rhythmic bursts of action potentials with a frequency and duration that were consistent with those of the spontaneous contractions. The periodic depolarizations were blocked by tetrodotoxin or Cd(2+). Surprisingly, the contractions and periodic depolarizations were insensitive to general blockade of synaptic transmission (by elevated Mg(2+) and reduced Ca(2+), or by Co(2+)) and to selective blockade of the major neurotransmitter receptors of the mature spinal cord (acetylcholine, GABA(A), NMDA, AMPA/kainate, and glycine). The periodic depolarizations were suppressed by heptanol or by intracellular acidification, treatments that are known to uncouple gap junctions, indicating that electrotonic synapses could underlie the earliest motor behavior. A few hours later, most motoneurons already showed a new pattern of repetitive activity consisting of bursts of glycinergic synaptic events, but these were not necessary for the spontaneous contractions. Transecting the spinal cord at the hindbrain border did not affect the rhythmic activity patterns of the motoneurons. We suggest that spontaneous contractions of the zebrafish embryo are mediated by an early spinal circuit that is independent of the main neurotransmitter systems and descending hindbrain projections that are required for locomotion in the mature vertebrate spinal cord. PMID- 10818132 TI - Muscarinic inhibition of calcium current and M current in Galpha q-deficient mice. AB - Activation of M(1) muscarinic acetylcholine receptors (M(1) mAChR) inhibits M type potassium currents (I(K(M))) and N-type calcium currents (I(Ca)) in mammalian sympathetic ganglia. Previous antisense experiments suggested that, in rat superior cervical ganglion (SCG) neurons, both effects were partly mediated by the G-protein Galpha(q) (Delmas et al., 1998a; Haley et al., 1998a), but did not eliminate a contribution by other pertussis toxin (PTX)-insensitive G proteins. We have tested this further using mice deficient in the Galpha(q) gene. PTX-insensitive M(1) mAChR inhibition of I(Ca) was strongly reduced in Galpha(q) /- mouse SCG neurons and was fully restored by acute overexpression of Galpha(q). In contrast, M(1) mAChR inhibition of I(K(M)) persisted in Galpha(q)-/- mouse SCG cells. However, unlike rat SCG neurons, muscarinic inhibition of I(K(M)) was partly PTX-sensitive. Residual (PTX-insensitive) I(K(M)) inhibition was slightly reduced in Galpha(q) -/- neurons, and the remaining response was then suppressed by anti-Galpha(q/11) antibodies. Bradykinin (BK) also inhibits I(K(M)) in rat SCG neurons via a PTX-insensitive G-protein (G(q) and/or G(11); Jones et al., 1995). In mouse SCG neurons, I(K(M)) inhibition by BK was fully PTX-resistant. It was unchanged in Galpha(q) -/- mice but was abolished by anti-Galpha(q/11) antibody. We conclude that, in mouse SCG neurons (1) M(1) mAChR inhibition of I(Ca) is mediated principally by G(q), (2) M(1) mAChR inhibition of I(K(M)) is mediated partly by G(q), more substantially by G(11), and partly by a PTX-sensitive G protein(s), and (3) BK-induced inhibition of I(K(M)) is mediated wholly by G(11). PMID- 10818133 TI - Role of cAMP cascade in synaptic stability and plasticity: ultrastructural and physiological analyses of individual synaptic boutons in Drosophila memory mutants. AB - Mutations of the genes rutabaga (rut) and dunce (dnc) affect the synthesis and degradation of cAMP, respectively, and disrupt learning in Drosophila. Combined ultrastructural analysis and focal electrophysiological recording in the larval neuromuscular junction revealed a loss of stability and fine tuning of synaptic structure and function in both mutants. Increased ratios of docked/undocked vesicles and poorly defined synaptic specializations characterized dnc synapses. In contrast, rut boutons possessed fewer, although larger, synapses with lower proportions of docked vesicles. At reduced Ca(2+) levels, decreased quantal content coupled with an increase in failure rate was seen in rut boutons and reduced pair-pulse facilitation were found in both rut and dnc mutants. At physiological Ca(2+) levels, strong enhancement, instead of depression, in evoked release was observed in some dnc and rut boutons during 10 Hz tetanus. Furthermore, increased variability of synaptic transmission, including fluctuation and asynchronicity of evoked release, paralleled an increase in synapse size variation in both dnc and rut boutons, which might impose problems for effective signal processing in the nervous system. Pharmacological and genetic studies indicated broader ranges of physiological alteration by dnc and rut mutations than either the acute effects of cAMP analogs or the available mutations that affect cAMP-dependent protein kinase (PKA) activity. This is consistent with previous reports of more severe learning defects in dnc and rut mutations than these PKA mutants and allows identification of the phenotypes involving long-term developmental regulation and those conferred by PKA. PMID- 10818134 TI - Inhibition of activity-dependent arc protein expression in the rat hippocampus impairs the maintenance of long-term potentiation and the consolidation of long term memory. AB - It is widely believed that the brain processes information and stores memories by modifying and stabilizing synaptic connections between neurons. In experimental models of synaptic plasticity, such as long-term potentiation (LTP), the stabilization of changes in synaptic strength requires rapid de novo RNA and protein synthesis. Candidate genes, which could underlie activity-dependent plasticity, have been identified on the basis of their rapid induction in brain neurons. Immediate-early genes (IEGs) are induced in hippocampal neurons by high frequency electrical stimulation that induces LTP and by behavioral training that results in long-term memory (LTM) formation. Here, we investigated the role of the IEG Arc (also termed Arg3.1) in hippocampal plasticity. Arc protein is known to be enriched in dendrites of hippocampal neurons where it associates with cytoskeletal proteins (Lyford et al., 1995). Arc is also notable in that its mRNA and protein accumulate in dendrites at sites of recent synaptic activity (Steward et al., 1998). We used intrahippocampal infusions of antisense oligodeoxynucleotides to inhibit Arc protein expression and examined the effect of this treatment on both LTP and spatial learning. Our studies show that disruption of Arc protein expression impairs the maintenance phase of LTP without affecting its induction and impairs consolidation of LTM for spatial water task training without affecting task acquisition or short-term memory. Thus, Arc appears to play a fundamental role in the stabilization of activity-dependent hippocampal plasticity. PMID- 10818135 TI - The evolution of lipophilin genes from invertebrates to tetrapods: DM-20 cannot replace proteolipid protein in CNS myelin. AB - The proteolipid protein (PLP) gene encodes two myelin-specific protein isoforms, DM-20 and PLP, which are members of the highly conserved lipophilin family of transmembrane proteins. While the functions of this family are poorly understood, the fact that null mutations of the PLP gene cause leukodystrophy in man is testament to the importance of DM-20 and PLP in normal CNS function. PLP differs from DM-20 by the presence of a 35 amino acid domain exposed to the cytoplasm, which is not encoded by other lipophilin genes and appears to have arisen in amphibians approximately 300 million years before present. However, the lipophilin gene family can be traced back at least 550 million years and is represented in Drosophila and silkworms. Thus, from an evolutionary perspective PLP can reasonably be anticipated to perform functions in CNS myelin that cannot be accomplished by other lipophilins. Herein we use a novel knock-in strategy to generate mice expressing wild-type levels of a Plp gene that has been modified to encode only DM-20. Although DM-20 is incorporated into functional compact myelin sheaths in young animals, our data show that the 35 amino acid PLP-specific peptide is required to engender the normal myelin period and to confer long-term stability on this multilamellar membrane. PMID- 10818136 TI - Intracellular modulation of NMDA receptor function by antipsychotic drugs. AB - The present study deals with the functional interaction of antipsychotic drugs and NMDA receptors. We show that both the conventional antipsychotic drug haloperidol and the atypical antipsychotic drug clozapine mediate gene expression via intracellular regulation of NMDA receptors, albeit to different extents. Data obtained in primary striatal culture demonstrate that the intraneuronal signal transduction pathway activated by haloperidol, the cAMP pathway, leads to phosphorylation of the NR1 subtype of the NMDA receptor at (897)Ser. Haloperidol treatment is likewise shown to increase (897)Ser-NR1 phosphorylation in rats in vivo. Mutation of (896)Ser and (897)Ser to alanine, which prevents phosphorylation at both sites, inhibits cAMP-mediated gene expression. We conclude that antipsychotic drugs have the ability to modulate NMDA receptor function by an intraneuronal signal transduction mechanism. This facilitation of NMDA activity is necessary for antipsychotic drug-mediated gene expression and may contribute to the therapeutic benefits as well as side effects of antipsychotic drug treatment. PMID- 10818137 TI - Cytoskeletal links of neuronal acetylcholine receptors containing alpha 7 subunits. AB - Nicotinic acetylcholine receptors serve a variety of signaling functions in the nervous system depending on cellular location, but little is known about mechanisms responsible for tethering them at specific sites. Among the most interesting are receptors containing the alpha7 gene product, because of their abundance and high relative permeability to calcium. On chick ciliary ganglion neurons alpha7-containing receptors are highly concentrated on somatic spines folded into discrete patches on the cell. We show that the spines contain filamentous actin and drebrin. After cell dissociation, the actin slowly redistributes, the spines retract, and the alpha7-containing receptors disperse and are subsequently lost from the surface. Latrunculin A, a drug that depolymerizes filamentous actin, accelerates receptor dispersal, whereas jasplikinolide, a drug that stabilizes the actin cytoskeleton, preserves large receptor clusters and prevents receptor loss from the surface. The receptors are resistant to extraction by nonionic detergent even after latrunculin A treatment. Other, less abundant, nicotinic receptors on the neurons are readily solubilized by the detergent even though these receptors are located in part on the spines. The results demonstrate that the actin cytoskeleton is important for retaining receptor-rich spines and indicate that additional cytoskeletal elements or molecular interactions specific for alpha7-containing receptors influence their fate in the membrane. The cytoskeletal elements involved are not dependent on the architecture of the postsynaptic density because alpha7-containing receptors are excluded from such sites on ciliary ganglion neurons. PMID- 10818138 TI - cAMP response element-mediated gene transcription is upregulated by chronic antidepressant treatment. AB - Regulation of gene transcription via the cAMP-mediated second messenger pathway has been implicated in the actions of antidepressant drugs, but studies to date have not demonstrated such an effect in vivo. To directly study the regulation of cAMP response element (CRE)-mediated gene transcription by antidepressants, transgenic mice with a CRE-LacZ reporter gene construct were administered one of three different classes of antidepressants: a norepinephrine selective reuptake inhibitor (desipramine), a serotonin selective reuptake inhibitor (fluoxetine), or a monoamine oxidase inhibitor (tranylcypromine). Chronic, but not acute, administration of these antidepressants significantly increased CRE-mediated gene transcription, as well as the phosphorylation of CRE binding protein (CREB), in several limbic brain regions thought to mediate the action of antidepressants, including the cerebral cortex, hippocampus, amygdala, and hypothalamus. These results demonstrate that chronic antidepressant treatment induces CRE-mediated gene expression in a neuroanatomically differentiated pattern and further elucidate the molecular mechanisms underlying the actions of these widely used therapeutic agents. PMID- 10818139 TI - Functional consequences of reduction in NMDA receptor glycine affinity in mice carrying targeted point mutations in the glycine binding site. AB - We have used site-directed mutagenesis in conjunction with homologous recombination to generate two mouse lines carrying point mutations in the glycine binding site of the NMDAR1 subunit (Grin1). Glycine concentration-response curves from acutely dissociated hippocampal neurons revealed a 5- and 86-fold reduction in receptor glycine affinity in mice carrying Grin1(D481N) and Grin1(K483Q) mutations, respectively, whereas receptor glutamate affinity remained unaffected. Homozygous mutant Grin1(D481N) animals are viable and fertile and appear to develop normally. However, homozygous mutant Grin1(K483Q) animals are significantly lighter at birth, do not feed, and die within a few days. No gross abnormalities in CNS anatomy were detected in either Grin1(D481N) or Grin1(K483Q) mice. Interestingly, in situ hybridization and Western blot analysis revealed changes in the expression levels of NMDA receptor subunits in Grin1(D481N) mice relative to wild type that may represent a compensatory response to the reduction in receptor glycine affinity. Grin1(D481N) mice exhibited deficits in hippocampal theta burst-induced long-term potentiation (LTP) and spatial learning and also a reduction in sensitivity to NMDA-induced seizures relative to wild-type controls, consistent with a reduced activation of NMDA receptors. Mutant mice exhibited normal prepulse inhibition but showed increased startle reactivity. Preliminary analysis indicated that the mice exhibit a decreased natural aversion to an exposed environment. The lethal phenotype of Grin1(K483Q) animals confirms the critical role of NMDA receptor activation in neonatal survival. A milder reduction in receptor glycine affinity results in an impairment of LTP and spatial learning and alterations in anxiety-related behavior, providing further evidence for the role of NMDA receptor activation in these processes. PMID- 10818140 TI - High-level neuronal expression of abeta 1-42 in wild-type human amyloid protein precursor transgenic mice: synaptotoxicity without plaque formation. AB - Amyloid plaques are a neuropathological hallmark of Alzheimer's disease (AD), but their relationship to neurodegeneration and dementia remains controversial. In contrast, there is a good correlation in AD between cognitive decline and loss of synaptophysin-immunoreactive (SYN-IR) presynaptic terminals in specific brain regions. We used expression-matched transgenic mouse lines to compare the effects of different human amyloid protein precursors (hAPP) and their products on plaque formation and SYN-IR presynaptic terminals. Four distinct minigenes were generated encoding wild-type hAPP or hAPP carrying mutations that alter the production of amyloidogenic Abeta peptides. The platelet-derived growth factor beta chain promoter was used to express these constructs in neurons. hAPP mutations associated with familial AD (FAD) increased cerebral Abeta(1-42) levels, whereas an experimental mutation of the beta-secretase cleavage site (671(M-->I)) eliminated production of human Abeta. High levels of Abeta(1-42) resulted in age-dependent formation of amyloid plaques in FAD-mutant hAPP mice but not in expression-matched wild-type hAPP mice. Yet, significant decreases in the density of SYN-IR presynaptic terminals were found in both groups of mice. Across mice from different transgenic lines, the density of SYN-IR presynaptic terminals correlated inversely with Abeta levels but not with hAPP levels or plaque load. We conclude that Abeta is synaptotoxic even in the absence of plaques and that high levels of Abeta(1-42) are insufficient to induce plaque formation in mice expressing wild-type hAPP. Our results support the emerging view that plaque-independent Abeta toxicity plays an important role in the development of synaptic deficits in AD and related conditions. PMID- 10818141 TI - Neurotrophin-3 sorts to the constitutive secretory pathway of hippocampal neurons and is diverted to the regulated secretory pathway by coexpression with brain derived neurotrophic factor. AB - Hippocampal neurons release nerve growth factor (NGF) through the constitutive secretory pathway, thus allowing the protein to be continuously available for promoting nerve cell survival. In contrast, hippocampal neurons use the regulated secretory pathway to process brain-derived neurotrophic factor (BDNF), which alters synaptic activity when released acutely from dense-core vesicles. Thus, understanding how neurons sort and deliver neurotrophins may provide clues to their functions in brain. In this study, we monitored the processing and delivery of neurotrophin-3 (NT-3). Pulse-chase studies, immunocytochemistry, and secretagogue-induced release experiments were performed on cultured hippocampal neurons and AtT-20 cells infected with vaccinia viruses encoding the NT-3 precursor (pro-NT-3). Results show that most newly synthesized NT-3 is released through the constitutive secretory pathway as a result of furin-mediated endoproteolytic cleavage of pro-NT-3 in the trans-Golgi network. Pro-NT-3 can also be diverted into the regulated secretory pathway when cells are treated with alpha1-PDX, a selective inhibitor of furin-like enzymes, or when pro-NT-3 expression is increased by transient transfection methods. In cells coinfected with viruses coding for pro-NT-3 and pro-BDNF, NT-3 is sorted into the regulated pathway, stored in secretory granules, and released in response to extracellular cues together with BDNF, apparently as a result of heterodimerization, as suggested by coimmunoprecipitation data. Taken together, these data show that sorting of the NT-3 precursor can occur in both the constitutive and regulated secretory pathways, which is consistent with NT-3 having both survival-promoting and synapse-altering functions. PMID- 10818142 TI - Identification of proteins in the postsynaptic density fraction by mass spectrometry. AB - Our understanding of the organization of postsynaptic signaling systems at excitatory synapses has been aided by the identification of proteins in the postsynaptic density (PSD) fraction, a subcellular fraction enriched in structures with the morphology of PSDs. In this study, we have completed the identification of most major proteins in the PSD fraction with the use of an analytical method based on mass spectrometry coupled with searching of the protein sequence databases. At least one protein in each of 26 prominent protein bands from the PSD fraction has now been identified. We found 7 proteins not previously known to be constituents of the PSD fraction and 24 that had previously been associated with the PSD by other methods. The newly identified proteins include the heavy chain of myosin-Va (dilute myosin), a motor protein thought to be involved in vesicle trafficking, and the mammalian homolog of the yeast septin protein cdc10, which is important for bud formation in yeast. Both myosin-Va and cdc10 are threefold to fivefold enriched in the PSD fraction over brain homogenates. Immunocytochemical localization of myosin-Va in cultured hippocampal neurons shows that it partially colocalizes with PSD-95 at synapses and is also diffusely localized in cell bodies, dendrites, and axons. Cdc10 has a punctate distribution in cell bodies and dendrites, with some of the puncta colocalizing with PSD-95. The results support a role for myosin-Va in transport of materials into spines and for septins in the formation or maintenance of spines. PMID- 10818143 TI - Ciliary neurotrophic factor and stress stimuli activate the Jak-STAT pathway in retinal neurons and glia. AB - Ciliary neurotrophic factor (CNTF) is pleiotrophic for central, peripheral, and sensory neurons. In the mature retina, CNTF treatment enhances survival of retinal ganglion and photoreceptor cells exposed to otherwise lethal perturbation. To understand its mechanism of action in vivo, the adult rat retina was used as a model to investigate CNTF-mediated activation of Janus kinase/signal transducer and activator of transcription (Jak-STAT) and ras mitogen activated protein kinase (ras-MAPK). Intravitreal injection of Axokine, an analog of CNTF, phosphorylates STAT3 and MAPK and produces delayed upregulation of total STAT3 and STAT1 protein in rat retina. Activated STAT3 is predominantly localized in nuclei of retinal Muller (glial) cells, ganglion cells, and astrocytes, but not in photoreceptors. Although CNTF alpha-receptor (CNTFRalpha) mRNA and protein are localized predominantly if not exclusively in retinal neurons, coincident CNTF-mediated STAT3 signaling was observed in both glia and neurons. CNTF-induced activation of Jak-STAT signaling prompted us to investigate STAT3 phosphorylation after a variety of stress-mediated, conditioning stimuli. We show that STAT3 is activated in the retina after exposure to subtoxic bright light, mechanical trauma, and systemic administration of the alpha(2)-adrenergic agonist xylazine, all of which have been shown previously to condition photoreceptors to resist light-induced degeneration. These results demonstrate that CNTF directly stimulates Jak-STAT and ras-MAPK cascades in vivo and strongly suggest that STAT3 signaling is an underlying component of neural responsiveness to stress stimuli. The observation that CNTF activates STAT3 in ganglion cells, but not in photoreceptors, suggests that Jak STAT signaling influences neuronal survival via both direct and indirect modes of action. PMID- 10818144 TI - Functional coupling between neurons and glia. AB - Neuronal-glial interactions play an important role in information processing in the CNS. Previous studies have indicated that electrotonic coupling between locus ceruleus (LC) neurons is involved in synchronizing the spontaneous activity. The results of the present study extend the functional electrotonic coupling to interactions between neurons and glia. Spontaneous oscillations in the membrane potential were observed in a subset of glia. These oscillations were synchronous with the firing of neurons, insensitive to transmitter receptor antagonists and disrupted by carbenoxolone, a gap junction blocker. Hyperpolarization of neurons with [Met] (5)enkephalin blocked the oscillations in glia. Selective depolarization of glia with a glutamate transporter substrate (l-alpha aminoadipic acid) increased the neuronal firing rate, suggesting that changes in the membrane potential of glia can modulate neuronal excitability through heterocellular coupling. Dye-coupling experiments further confirmed that small molecules could be transferred through gap junctions between these distinct cell types. No dye transfer was observed between neurons and oligodendrocytes or between astrocytes and oligodendrocytes, suggesting that the junctional communication was specific for astrocytes and neurons. Finally, immunoelectron microscopy studies established that connexins, the proteins that form gap junctions, were present on portions of the plasmalemma, bridging the cytoplasm of neurons and glia in LC. This heterocellular coupling extends the mechanisms by which glia participate in the network properties of the LC in which the degree of coupling is thought to influence cognitive performance. PMID- 10818145 TI - Heparan sulfate heterogeneity in skeletal muscle basal lamina: demonstration by phage display-derived antibodies. AB - The basal lamina (BL) enveloping skeletal muscle fibers contains different glycoproteins, including proteoglycans. To obtain more information on the glycosaminoglycan moiety of proteoglycans, we have selected a panel of anti heparan sulfate (HS) antibodies from a semisynthetic antibody phage display library by panning against glycosaminoglycan preparations derived from skeletal muscle. Epitope recognition by the antibodies is strongly dependent on O- and N sulfation of the heparan sulfate. Immunostaining with these antibodies showed a distinct distribution of heparan sulfate epitopes in muscle basal lamina of various species. Clear differences in staining intensity were observed between neural, synaptic, and extrasynaptic basal laminae. Moreover, temporal and regional changes in abundancy of heparan sulfate epitopes were observed during muscle development both in vitro and in vivo. Taken together, these data suggest a role for specific heparan sulfate domains/species in myogenesis and synaptogenesis. Detailed analysis of the functions of heparan sulfate epitopes in muscle morphogenesis has now become feasible with the isolation of antibodies specific for distinct heparan sulfate epitopes. PMID- 10818146 TI - Local directional cues control growth polarity of dopaminergic axons along the rostrocaudal axis. AB - The vertebrate CNS is composed of a variety of longitudinal axonal tracts extending rostrally and caudally. Although recent studies have demonstrated that chemoattraction and chemorepulsion play key roles in axon guidance along the circumferential axis in the neural tube of the vertebrate, mechanisms of axonal elongation along the longitudinal axis, and most importantly, what determines rostrocaudal polarity of axonal growth, remains unknown. Here, we examined the mechanism that guides midbrain dopaminergic axons rostrally, using flat whole mount preparations of embryonic rat brain both in vivo and in vitro. At embryonic day 11 (E11) and early stage E12, dopaminergic neurons in the ventral midbrain extended short axons dorsally. By middle stage E12, these axons had increased in number, some deflecting rostrally and others caudally. At E13, almost all axons showed rostrally oriented growth heading toward the forebrain targets. In in vitro whole-mount preparations prepared from an E12 embryo and cultured for 24 hr, these axons showed rostrally oriented growth, but when they were forced to grow on substratum of reversed rostrocaudal polarity, they turned abruptly and grew following the polarity of the reversed midbrain substratum. These results suggest that local directional cues in the midbrain guide these axons rostrally and support the idea that substratum-associated polarized cues play an important role in axon guidance along the longitudinal axis. PMID- 10818147 TI - Uncoupling of myelin assembly and schwann cell differentiation by transgenic overexpression of peripheral myelin protein 22. AB - We have generated previously transgenic rats that overexpress peripheral myelin protein 22 (PMP22) in Schwann cells. In the nerves of these animals, Schwann cells have segregated with axons to the normal 1:1 ratio but remain arrested at the promyelinating stage, apparently unable to elaborate myelin sheaths. We have examined gene expression of these dysmyelinating Schwann cells using semiquantitative reverse transcription-PCR and immunofluorescence analysis. Unexpectedly, Schwann cell differentiation appears to proceed normally at the molecular level when monitored by the expression of mRNAs encoding major structural proteins of myelin. Furthermore, an aberrant coexpression of early and late Schwann cell markers was observed. PMP22 itself acquires complex glycosylation, suggesting that trafficking of the myelin protein through the endoplasmic reticulum is not significantly impaired. We suggest that PMP22, when overexpressed, accumulates in a late Golgi-cell membrane compartment and uncouples myelin assembly from the underlying program of Schwann cell differentiation. PMID- 10818148 TI - Granule cells and cerebellar boundaries: analysis of Unc5h3 mutant chimeras. AB - Mutations in the Unc5h3 gene, a receptor for the netrin 1 ligand, result in abnormal migrations of both Purkinje and granule cells to regions outside the cerebellum and of granule cells to regions within the cerebellum. Because both Purkinje and granule cells express this molecule, we sought to determine whether one or both of these cell types are the primary target of the mutation. Chimeric mice were made between wild-type ROSA26 transgenic mouse embryos (whose cells express beta-galactosidase) and Unc5h3 mutant embryos. The resulting chimeric brains exhibited a range of phenotypes. Chimeras that had a limited expression of the extracerebellar phenotype (movement of cerebellar cells into the colliculus and midbrain tegmentum) and the intracerebellar phenotype (migration of granule cells into white matter) had a normal-appearing cerebellum, whereas chimeras that had more ectopic cells had attenuated anterior cerebellar lobules. Furthermore, the colonization of colliculus and midbrain tegmentum by cerebellar cells was not equivalent in all chimeras, suggesting different origins for extracerebellar ectopias in these regions. The granule cells of the extracerebellar ectopias were almost entirely derived from Unc5h3/Unc5h3 mutant embryos, whereas the ectopic Purkinje cells were a mixture of both mutant and wild-type cells. Intracerebellar ectopias in the chimera were composed exclusively of mutant granule cells. These findings demonstrate that both inside and outside the cerebellum, the granule cell is the key cell type to demarcate the boundaries of the cerebellum. PMID- 10818149 TI - The small GTP-binding protein TC10 promotes nerve elongation in neuronal cells, and its expression is induced during nerve regeneration in rats. AB - We have made a rat cDNA library using nerve-transected hypoglossal nuclei. Using this library, we performed expressed-sequence tag analysis coupled with in situ hybridization to identify genes whose expression is altered in response to nerve injury. In this gene screening, a member of Rho family GTPases, TC10, which had not yet been characterized in neuronal cells, was identified. TC10 mRNA expression was very low in normal motor neurons; however, axotomy induced its expression dramatically. Other family members such as RhoA, Rac1, and Cdc42 were moderately expressed in normal motor neurons and showed slight upregulation after axotomy. The expression level of TC10 mRNA was low in the embryonic brain and gradually increased with development. However, the expression of TC10 mRNA in the adult brain was lower and more restricted than that of RhoA, Rac1, and Cdc42. Cultured dorsal root ganglia exhibited dramatic neurite extension secondary to adenovirus-mediated expression of TC10. It can be concluded that although TC10 expression is lower in developing and mature motor neurons compared with other Rho family members, TC10 expression is induced by nerve injury to play a crucial role in nerve regeneration, particularly neurite elongation, in cooperation with other family members. PMID- 10818150 TI - A-type K+ current mediated by the Kv4 channel regulates the generation of action potential in developing cerebellar granule cells. AB - During neuronal differentiation and maturation, electrical excitability is essential for proper gene expression and the formation of synapses. The expression of ion channels is crucial for this process; in particular, voltage gated K(+) channels function as the key determinants of membrane excitability. Previously, we reported that the A-type K(+) current (I(A)) and Kv4.2 K(+) channel subunit expression increased in cultured cerebellar granule cells with time. To examine the correlation between ion currents and the action potential, in the present study, we measured developmental changes of action potentials in cultured granule cells using the whole-cell patch-clamp method. In addition to an observed increment of I(A), we found that the Na(+) current also increased during development. The increase in both currents was accompanied by a change in the membrane excitability from the nonspiking type to the repetitive firing type. Next, to elucidate whether Kv4.2 is responsible for the I(A) and to assess the effect of Kv4 subunits on action potential waveform, we transfected a cDNA encoding a dominant-negative mutant Kv4.2 (Kv4.2dn) into cultured cells. Expression of Kv4.2dn resulted in the elimination of I(A) in the granule cells. This result demonstrates that members of the Kv4 subfamily are responsible for the I(A) in developing granule cells. Moreover, elimination of I(A) resulted in shortening of latency before the first spike generation. In contrast, expression of wild-type Kv4.2 resulted in a delay in latency. This indicates that appearance of I(A) is critically required for suppression of the excitability of granule cells during their maturation. PMID- 10818151 TI - Role of founder cell deficit and delayed neuronogenesis in microencephaly of the trisomy 16 mouse. AB - Development of the neocortex of the trisomy 16 (Ts16) mouse, an animal model of Down syndrome (DS), is characterized by a transient delay in the radial expansion of the cortical wall and a persistent reduction in cortical volume. Here we show that at each cell cycle during neuronogenesis, a smaller proportion of Ts16 progenitors exit the cell cycle than do control, euploid progenitors. In addition, the cell cycle duration was found to be longer in Ts16 than in euploid progenitors, the Ts16 growth fraction was reduced, and an increase in apoptosis was observed in both proliferative and postmitotic zones of the developing Ts16 neocortical wall. Incorporation of these changes into a model of neuronogenesis indicates that they are sufficient to account for the observed delay in radial expansion. In addition, the number of neocortical founder cells, i.e., precursors present just before neuronogenesis begins, is reduced by 26% in Ts16 mice, leading to a reduction in overall cortical size at the end of Ts16 neuronogenesis. Thus, altered proliferative characteristics during Ts16 neuronogenesis result in a delay in the generation of neocortical neurons, whereas the founder cell deficit leads to a proportional reduction in the overall number of neurons. Such prenatal perturbations in either the timing of neuron generation or the final number of neurons produced may lead to significant neocortical abnormalities such as those found in DS. PMID- 10818152 TI - Insulin-like growth factor I stimulates dendritic growth in primary somatosensory cortex. AB - The temporal and spatial distributions of several growth factors suggest roles in the regulation of neuronal differentiation in the neocortex. Among such growth factors, the insulin-like growth factors (IGF-I and -II) are of particular interest because they are available to neurons from multiple sources under independent control. IGF-I is produced by many neurons throughout the brain and also by cells in the cerebral vasculature. IGF-II is found at high levels in the CSF, and both IGF-I and IGF-II cross the blood-brain barrier. Thus, the IGFs may act as both paracrine and endocrine regulators of neuronal development. As an initial step toward understanding the influence of IGFs in the developing cerebral cortex, the present study examined the effects of IGF-I and of the neurotrophins brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3) on the dendritic complexity of layer 2 pyramidal neurons. The results demonstrate that IGF-I increased the branching and total extent of both apical and basal dendrites of pyramidal cells in organotypic slices of rat primary somatosensory cortex. BDNF and NT-3 also enhanced dendritic development, but the two neurotrophins increased the extent of only basal, not apical, dendrites and promoted greater elongation than was seen after IGF-I treatment. These results provide direct evidence that IGF-I can regulate the dendritic elaboration of cortical neurons and indicate that endogenous IGFs may influence dendritic differentiation and the formation of cortical connections. In addition, IGF dependent regulation of dendritic structure may represent a link between age related declines in IGFs and cognitive deficits seen in senescence. PMID- 10818153 TI - A MAP kinase-signaling pathway mediates neurite outgrowth on L1 and requires Src dependent endocytosis. AB - The neural cell adhesion molecule L1 mediates the axon outgrowth, adhesion, and fasciculation necessary for proper development of synaptic connections. Mutations of human L1 cause an X-linked mental retardation syndrome termed CRASH (corpus callosum hypoplasia, retardation, aphasia, spastic paraplegia, and hydrocephalus), and L1 knock-out mice display defects in neuronal process extension resembling the CRASH phenotype. Little is known about the biochemical or cellular mechanism by which L1 performs neuronal functions. Here it is demonstrated that clustering of L1 with antibodies or L1 protein in rodent B35 neuroblastoma and cerebellar neuron cultures induced the phosphorylation/activation of the mitogen-activated protein kinases (MAPKs) and extracellular signal-regulated kinases 1 and 2. MAPK activation was essential for L1-dependent neurite outgrowth, because chemical inhibitors [2-(2'-amino-3' methoxyphenyl)-oxanaphthalen-4-one and 1,4-diamino-2, 3-dicyano-1,4-bis(2 aminophenylthio)butadiene] of the MAPK kinase MEK strongly suppressed neurite outgrowth by cerebellar neurons on L1. The nonreceptor tyrosine kinase pp60(c src) was required for L1-triggered MAPK phosphorylation, as shown in src-minus cerebellar neurons and by expression of the kinase-inactive mutant Src(K295M) in B35 neuroblastoma cells. Phosphatidylinositol 3-kinase (PI3-kinase) and the small GTPase p21(rac) were identified as signaling intermediates to MAPK by phosphoinositide and Rac-GTP assays and expression of inhibitory mutants. Antibody-induced endocytosis of L1, visualized by immunofluorescence staining and confocal microscopy of B35 cells, was blocked by expression of kinase-inactive Src(K295M) and dominant-negative dynamin(K44A) but not by inhibitors of MEK or PI3-kinase. Dynamin(K44A) also inhibited L1 antibody-triggered MAPK phosphorylation. This study supports a model in which pp60(c-src) regulates dynamin-mediated endocytosis of L1 as an essential step in MAPK-dependent neurite outgrowth on an L1 substrate. PMID- 10818154 TI - The development of abnormal axon trajectories after rotation of one eye in Xenopus. AB - The targeting of isthmotectal axons in the Xenopus binocular pathway is guided by both activity-dependent cues and activity-independent cues. Abnormal visual activity induced by unilateral eye rotation overrides activity-independent cues and causes isthmotectal axons to arborize at new locations during a critical period of development that ends approximately 3 months postmetamorphosis (PM). Horseradish peroxidase staining of isthmotectal axons reveals that they normally run rostrocaudally in the tectum; in contrast, those axons in animals with early eye rotation have circuitous trajectories. In this paper, by studying the trajectories and branching patterns of isthmotectal axons at different times after eye rotation, we aimed to investigate when and how activity cues determine the projection pattern of isthmotectal axons. As suggested by electrophysiological recording, isthmotectal axons initially grow normally and make arbors according to activity-independent cues despite the presence of abnormal visual input. Our findings demonstrate that the development of abnormal trajectories starts by 2 weeks PM in response to eye rotation and is a protracted process. It begins in the tectal regions in which the initial connections of isthmotectal axons are first formed according to activity-independent cues. At transitional stages (5 and 10 weeks), axons with arbors at two different locations are observed, with locations corresponding to the old and new termination sites, respectively. Later, at 10 weeks of age, the fainter horseradish peroxidase staining in arbors at old termination sites suggests that the older arbors are undergoing withdrawal. PMID- 10818155 TI - Leukemia inhibitory factor requires concurrent p75LNTR signaling to induce apoptosis of cultured sympathetic neurons. AB - Apoptosis may result either from positive induction by ligand binding to a plasma membrane receptor or from negative induction attributable to loss of a suppressor signal. For example, apoptosis of developing sympathetic neurons may be induced in culture either by exposure to leukemia inhibitory factor (LIF) or by deprivation of nerve growth factor. This study compared the cell death pathways activated in sympathetic neurons by these two different stimuli. Both types of cell death were developmentally regulated; both were maximal in the immediate postnatal period and disappeared over the next 2 weeks. Both types of cell death were reduced by genetic deletion of Bax or by virally mediated overexpression of Bcl-2. Similarly both were reduced by inhibition of caspase activity or by inhibition of Nedd-2 synthesis with antisense oligonucleotides. Finally, both involved activation of c-Jun N-terminal kinase (JNK) signaling. Nedd-2 expression by sympathetic neurons declined in parallel with the developmental loss of LIF mediated cell death, suggesting that downregulation of the caspase during development may underlie the loss of cytokine-mediated apoptosis. Treatment of sympathetic neurons with an antibody that blocks the function of the low-affinity neurotrophin receptor (p75(LNTR)) prevented LIF-induced cell death. Similarly genetic deletion of p75(LNTR) prevented apoptosis after LIF treatment. These observations suggest that concurrent p75(LNTR) signaling is necessary for LIF induced cell death and that cytokine-mediated cell death and growth factor deprivation appear to activate the same intracellular pathways involving JNK signaling. PMID- 10818156 TI - Upregulation of cAMP response element-mediated gene expression during experience dependent plasticity in adult neocortex. AB - Gene transcription is thought to be essential for memory consolidation and long lasting changes in synaptic function. In particular, the signal transduction pathways that activate the transcription factor cAMP response element binding protein (CREB) have been implicated in the process of synaptic potentiation. To study the involvement of this pathway in neocortical plasticity within the barrel cortex, we have used a strain of mice carrying a LacZ reporter gene with six cAMP response elements (CREs) upstream of a minimal promoter. Removal of all but one facial whisker results in the expansion of the spared whisker's functional representation within somatosensory cortex. Under the same conditions of whisker deprivation, we observed a strong (eightfold compared with baseline) and highly place-specific upregulation of CRE-mediated gene transcription in layer IV of the spared whisker barrel. Reporter gene upregulation occurred rapidly after deprivation (16 hr) and was only observed under experimental conditions capable of inducing whisker response potentiation. LacZ expression in layer IV was accompanied by an increase in responsiveness of a subpopulation of layers II/III cells to spared whisker stimulation as determined by in vivo single-unit recording. Given that CREB is involved in the expression of plasticity in superficial layers (Glazewski et al., 1999), and yet CRE-mediated gene expression occurs in layer IV, it is likely that the molecular events initiating plasticity occur presynaptically to the cells that exhibit changes in their receptive field properties. PMID- 10818157 TI - Role and origin of the GABAergic innervation of dorsal raphe serotonergic neurons. AB - Extracellular electrophysiological recordings in freely moving cats have shown that serotonergic neurons from the dorsal raphe nucleus (DRN) fire tonically during wakefulness, decrease their activity during slow wave sleep (SWS), and are nearly quiescent during paradoxical sleep (PS). The mechanisms at the origin of the modulation of activity of these neurons are still unknown. Here, we show in the unanesthetized rat that the iontophoretic application of the GABA(A) antagonist bicuculline on dorsal raphe serotonergic neurons induces a tonic discharge during SWS and PS and an increase of discharge rate during quiet waking. These data strongly suggest that an increase of a GABAergic inhibitory tone present during wakefulness is responsible for the decrease of activity of the dorsal raphe serotonergic cells during slow wave and paradoxical sleep. In addition, by combining retrograde tracing with cholera toxin B subunit and glutamic acid decarboxylase immunohistochemistry, we demonstrate that the GABAergic innervation of the dorsal raphe nucleus arises from multiple distant sources and not only from interneurons as classically accepted. Among these afferents, GABAergic neurons located in the lateral preoptic area and the pontine ventral periaqueductal gray including the DRN itself could be responsible for the reduction of activity of the serotonergic neurons of the dorsal raphe nucleus during slow wave and paradoxical sleep, respectively. PMID- 10818158 TI - Vertical shifts in self-administration dose-response functions predict a drug vulnerable phenotype predisposed to addiction. AB - The role of individual differences in the etiology of addiction is a very controversial issue. Neuroendocrine phenotypes that are able to predispose an individual to the development of drug intake have been identified previously. However, such information has been gathered by comparing individuals who differ in their sensitivity to low doses of the drug. Consequently, it remains unclear whether a phenotype predicting a higher sensitivity to low drug doses would be relevant in environmental conditions, such as the ones encountered by humans in which high drug doses are available. In this report, we studied dose-response, dose-intake, and ratio-intake functions for intravenous cocaine self administration in the laboratory rat. We show that individual differences in drug self-administration originate from vertical shift in the dose-response function. Thus, no matter the dose, drug intake is very high in some "vulnerable" subjects and very low in other "resistant" ones. Vulnerable subjects, the upward shifted ones, would then have a higher chance to develop drug abuse also when high drug doses are available. In conclusion, these results provide a solid foundation for the existence of a drug-vulnerable phenotype relevant for the etiology of addiction. PMID- 10818159 TI - The role of the hippocampus in instrumental conditioning. AB - Considerable evidence suggests that, in instrumental conditioning, rats can encode both the specific action-outcome associations to which they are exposed and the degree to which an action is causal in producing its associated outcome. Three experiments assessed the involvement of the hippocampus in encoding these aspects of instrumental learning. In each study, rats with electrolytic lesions of the dorsal hippocampus and sham-lesioned controls were trained while hungry to press two levers, each of which delivered a unique food outcome. Experiments 1A and 1B used an outcome devaluation procedure to assess the effects of the lesion on encoding the action-outcome relationship. After training, one of the two outcomes was devalued using a specific satiety procedure, after which performance on the two levers was assessed in a choice extinction test. The lesion had no detectable effect on either the acquisition of instrumental performance or on the rats' sensitivity to outcome devaluation; lesion and sham groups both reduced responding on the lever associated with the devalued outcome compared with the other lever. In experiment 2, the sensitivity of hippocampal rats to the causal efficacy of their actions was assessed by selectively degrading the contingency between one of the actions and its associated outcome. Whereas sham rats selectively reduced performance on the lever for which the action-outcome contingency had been degraded, hippocampal rats did not. These results suggest that, in instrumental conditioning, lesions of the dorsal hippocampus selectively impair the ability of rats to represent the causal relationship between an action and its consequences. PMID- 10818160 TI - Reflex excitability regulates prepulse inhibition. AB - Presentation of a weak stimulus, a prepulse, before a reflex-evoking stimulus decreases the amplitude of the reflex response relative to reflex amplitude evoked without a preceding prepulse. For example, presenting a brief tone before a trigeminal blink-eliciting stimulus significantly reduces reflex blink amplitude. A common explanation of such data are that sensory processing of the prepulse modifies reflex circuit behavior. The current study investigates the converse hypothesis that the intrinsic characteristics of the reflex circuit rather than prepulse processing determine prepulse modification of trigeminal and acoustic reflex blinks. Unilateral lesions of substantia nigra pars compacta neurons created rats with hyperexcitable trigeminal reflex blinks but normally excitable acoustic reflex blinks. In control rats, presentation of a prepulse reduced the amplitude of both trigeminal and acoustic reflex blinks. In 6-OHDA lesioned rats, however, the same acoustic prepulse facilitated trigeminal reflex blinks but inhibited acoustic reflex blinks. The magnitude of prepulse modification correlated with reflex excitability. Humans exhibited the same pattern of prepulse modification. An acoustic prepulse facilitated the trigeminal reflex blinks of subjects with hyperexcitable trigeminal reflex blinks caused by Parkinson's disease. The same prepulse inhibited trigeminal reflex blinks of age matched control subjects. Prepulse modification also correlated with trigeminal reflex blink excitability. These data show that reflex modification by a prepulse reflects the intrinsic characteristics of the reflex circuit rather than an external adjustment of the reflex circuit by the prepulse. PMID- 10818161 TI - Dopamine activates masculine sexual behavior independent of the estrogen receptor alpha. AB - Estrogen receptor alpha (ERalpha) is believed to be a critical part of the regulatory processes involved in normal reproduction and sexual behavior. However, in this study we show the ERalpha is not required for display of masculine sexual behavior. Male and female, ERalpha knock-out (ERalphaKO) and wild-type mice were gonadectomized and implanted with testosterone. Sexual behavior and social preferences were tested after injection of the dopamine agonist, apomorphine (APO), or vehicle. All wild-type mice showed normal masculine behavior, including mounts and pelvic thrusts in females, and ejaculation in males. In agreement with past reports, ERalphaKO mice, given vehicle, failed to show mating behavior. Yet, ERalphaKO males given APO showed masculine copulatory behavior and chemoinvestigatory behavior directed at females. ERalphaKO females, treated with APO, mounted and thrusted when tested with receptive females. HPLC revealed that wild-type and ERalphaKO mice had equivalent catecholamine content in brain regions associated with masculine sexual behavior. These data show that the ERalpha is not essential during development or adulthood for the expression of masculine sexual behavior in mice. Moreover, dopamine can activate sexual behavior via a mechanism that either acts on an ER other than ERalpha or via an estrogen-independent pathway. PMID- 10818162 TI - Evidence that separate neural circuits in the nucleus accumbens encode cocaine versus "natural" (water and food) reward. AB - Electrophysiological recording procedures were used to examine nucleus accumbens (Acb) cell firing in rats trained to press a lever on a multiple schedule [ fixed ratio (FR)1, FR1] for either two "natural" reinforcers (food and water), or a natural reinforcer and intravenous self-administration of cocaine. Of 180 cells recorded during water and food reinforcement (n = 13 rats), 77 neurons were classified as phasically active, exhibiting one of three well-defined types of patterned discharges relative to the reinforced-response (Carelli and Deadwyler, 1994). Of the 77 phasic cells, the majority (68%) showed similar types of patterned discharges across the two natural reinforcer conditions. In contrast, of 127 neurons recorded during water and cocaine reinforcement (n = 8 rats), only 5 of 60 phasically active cells (8%) exhibited similar types of patterned discharges relative to water- and cocaine-reinforced responding. The remaining 55 phasic cells (92%) displayed patterned discharges relative to the cocaine reinforced response (n = 26 cells), or relative to the water-reinforced response (n = 29 cells), but not both. For some rats (n = 3), food was substituted for water in the task. Again, the majority of phasic neurons (13 of 14 cells, 93%) exhibited nonoverlapping firing patterns across the drug and natural reinforcer conditions. These findings indicate that in the well-trained animal, cocaine activates a neural circuit in the Acb that is largely separate from the circuit that processes information about food and water reward. PMID- 10818163 TI - Membrane mechanisms underlying contrast adaptation in cat area 17 in vivo. AB - Contrast adaptation is a psychophysical phenomenon, the neuronal bases of which reside largely in the primary visual cortex. The cellular mechanisms of contrast adaptation were investigated in the cat primary visual cortex in vivo through intracellular recording and current injections. Visual cortex cells, and to a much less extent, dorsal lateral geniculate nucleus (dLGN) neurons, exhibited a reduction in firing rate during prolonged presentations of a high-contrast visual stimulus, a process we termed high-contrast adaptation. In a majority of cortical and dLGN cells, the period of adaptation to high contrast was followed by a prolonged (5-80 sec) period of reduced responsiveness to a low-contrast stimulus (postadaptation suppression), an effect that was associated, and positively correlated, with a hyperpolarization of the membrane potential and an increase in apparent membrane conductance. In simple cells, the period of postadaptation suppression was not consistently associated with a decrease in the grating modulated component of the evoked synaptic barrages (the F1 component). The generation of the hyperpolarization appears to be at least partially intrinsic to the recorded cells, because the induction of neuronal activity with the intracellular injection of current resulted in both a hyperpolarization of the membrane potential and a decrease in the spike response to either current injections or visual stimuli. Conversely, high-contrast visual stimulation could suppress the response to low-intensity sinusoidal current injection. We conclude that control of the membrane potential by intrinsic neuronal mechanisms contributes importantly to the adaptation of neuronal responsiveness to varying levels of contrast. This feedback mechanism, internal to cortical neurons, provides them with the ability to continually adjust their responsiveness as a function of their history of synaptic and action potential activity. PMID- 10818164 TI - Cellular mechanisms of long-lasting adaptation in visual cortical neurons in vitro. AB - The cellular mechanisms of spike-frequency adaptation during prolonged discharges and of the slow afterhyperpolarization (AHP) that follows, as occur in vivo with contrast adaptation, were investigated with intracellular recordings of cortical neurons in slices of ferret primary visual cortex. Intracellular injection of 2 Hz sinusoidal or constant currents for 20 sec resulted in a slow (tau = 1-10 sec) spike-frequency adaptation, the degree of which varied widely among neurons. Reducing either [Ca(2+)](o) or [Na(+)](o) reduced the rate of spike-frequency adaptation. After the prolonged discharge was a slow (12-75 sec) AHP that was associated with an increase in membrane conductance and a rightward shift in the discharge frequency versus injected current relationship. The reversal potential of the slow AHP was sensitive to changes in [K(+)](o), indicating that it was mediated by a K(+) current. Blockade of transmembrane Ca(2+) conductances did not reduce the slow AHP. In contrast, reductions of [Na(+)](o) reduced the slow AHP, even in the presence of pronounced Ca(2+) spikes. We suggest that the activation of Na(+)-activated and Ca(2+)-activated K(+) currents plays an important role in prolonged spike-frequency adaptation and therefore may contribute to contrast adaptation and other forms of adaptation in the visual system in vivo. PMID- 10818165 TI - Circadian and homeostatic control of rapid eye movement (REM) sleep: promotion of REM tendency by the suprachiasmatic nucleus. AB - The daily timing of rapid eye movement (REM) sleep reflects an interaction between the circadian pacemaker located in the suprachiasmatic nucleus of the hypothalamus (SCN) and a homeostatic process that induces compensatory REM sleep in response to REM sleep loss. Whether the circadian variation in REM sleep propensity is caused by active promotion, inhibition, or passive gating of REM sleep homeostasis by the SCN is unknown. To investigate these possibilities, compensatory responses to 24 hr REM sleep deprivation (RSD) were compared between SCN-lesioned (SCNx) and sham-lesioned rats at different times of day in constant dark. The attempts to enter REM sleep (REM tendency) increased during RSD in all rats and were modulated by circadian phase in sham-lesioned, but not SCNx rats. REM sleep homeostasis interacted with circadian time, such that REM tendency doubled during the rest phase in sham-lesioned rats relative to SCNx rats (F((6,93)) = 17.9; p = 0.0001). However, REM tendency was indistinguishable between SCNx and sham-lesioned rats during the activity phase, suggesting the SCN does not inhibit REM tendency at this time. By contrast, the amount of compensatory REM sleep examined 2, 6, 12, or 24 hr after RSD did not depend on circadian phase. Thus, transitions into REM sleep are facilitated by the SCN during the rest phase, but the amount of REM sleep, once initiated, is determined primarily by homeostatic mechanisms. This work supports a role for the SCN in the active promotion of REM sleep at specific times of day. PMID- 10818166 TI - Control of response selection by reinforcer value requires interaction of amygdala and orbital prefrontal cortex. AB - Goal-directed actions are guided by expected outcomes of those actions. Humans with bilateral damage to ventromedial prefrontal cortex, or the amygdala, are deficient in their ability to use information about positive and negative outcomes to guide their choice behavior. Similarly, rats and monkeys with orbital prefrontal or amygdala damage have been found to be impaired in their responses to changing values of outcomes. In the present study, we tested whether direct, functional interaction between the amygdala and the orbital prefrontal cortex is necessary for guiding behavior based on expected outcomes. Unlike control monkeys, rhesus monkeys with surgical disconnection of these two structures, achieved by crossed unilateral lesions of the amygdala in one hemisphere and orbital prefrontal cortex in the other, combined with forebrain commissurotomy, were unable to adjust their choice behavior after a change in the outcome (here, a reduction in the value of a particular reinforcer). The lesions did not affect motivation to work for a food reinforcer, or food preferences, per se. Hence, the amygdala and orbital prefrontal cortex act as part of an integrated neural system guiding decision-making and adaptive response selection. PMID- 10818167 TI - Medial frontal cortex mediates perceptual attentional set shifting in the rat. AB - If rodents do not display the behavioral complexity that is subserved in primates by prefrontal cortex, then evolution of prefrontal cortex in the rat should be doubted. Primate prefrontal cortex has been shown to mediate shifts in attention between perceptual dimensions of complex stimuli. This study examined the possibility that medial frontal cortex of the rat is involved in the shifting of perceptual attentional set. We trained rats to perform an attentional set shifting task that is formally the same as a task used in monkeys and humans. Rats were trained to dig in bowls for a food reward. The bowls were presented in pairs, only one of which was baited. The rat had to select the bowl in which to dig by its odor, the medium that filled the bowl, or the texture that covered its surface. In a single session, rats performed a series of discriminations, including reversals, an intradimensional shift, and an extradimensional shift. Bilateral lesions by injection of ibotenic acid in medial frontal cortex resulted in impairment in neither initial acquisition nor reversal learning. We report here the same selective impairment in shifting of attentional set in the rat as seen in primates with lesions of prefrontal cortex. We conclude that medial frontal cortex of the rat has functional similarity to primate lateral prefrontal cortex. PMID- 10818168 TI - Toward understanding the biology of a complex phenotype: rat strain and substrain differences in the sensorimotor gating-disruptive effects of dopamine agonists. AB - Sensorimotor gating, measured by prepulse inhibition (PPI) of the startle reflex, is reduced in schizophrenia patients and in rats treated with dopamine agonists. Strain differences in the sensitivity to the PPI-disruptive effects of dopamine agonists may provide insight into the genetic basis for human population differences in sensorimotor gating. We reported strain differences in the sensitivity to the PPI-disruptive effects of the D1/D2 agonist apomorphine in adult rats, with greater sensitivity in Harlan Sprague Dawley (SDH) versus Wistar (WH) rats. However, Kinney et al. (1999) recently reported opposite findings, using Bantin-Kingman Sprague Dawley (SDBK) and Wistar (WBK) rats; in fact, SDBK rats did not exhibit clear apomorphine-induced reductions in sensorimotor gating. These new findings of Kinney et al. (1999) directly conflict with over 15 years of results from our laboratories and challenge interpretations from a large body of literature. The present studies carefully assessed drug effects on sensorimotor gating in SD versus W strains, across rat suppliers (H vs BK). Significantly greater SDH than WH apomorphine sensitivity in PPI measures was observed in both adult and 18 d pups, confirming that these strain differences are both robust and innate. These strain differences in apomorphine sensitivity were not found in adult BK rats. Supplier differences in sensitivity (SDH > SDBK) were also evident in the PPI-disruptive effects of D1 but not D2-family agonists; PPI was clearly disrupted by quinpirole in both SDH and SDBK rats. These findings demonstrate robust, innate, neurochemically specific, and apparently heritable phenotypic differences in an animal model of sensorimotor gating deficits in human neuropsychiatric disorders. PMID- 10818169 TI - Mutations of the caenorhabditis elegans brain-specific inorganic phosphate transporter eat-4 affect habituation of the tap-withdrawal response without affecting the response itself. AB - The studies reported here were designed to investigate the role of the mutation eat-4 in the response to tap and in habituation in the nematode Caenorhabditis elegans. In C. elegans eat-4 has been found to affect a number of glutamatergic pathways. It has been hypothesized to positively regulate glutaminase activity and therefore glutamatergic neurotransmission. In the eat-4(ky5) loss-of-function worms, there is presumably insufficient glutamate available for sustained transmission. In the experiments reported here eat-4 worms showed no differences from wild-type in the magnitude of response to a single tap, indicating that the neural circuit underlying the response was intact and functional in the mutant worms. However, when eat-4 worms were given repeated taps the resulting habituation was different from that seen in wild-type worms: eat-4 worms habituate more rapidly and recover more slowly than wild-type worms at all interstimulus intervals tested. In addition, eat-4 worms do not show dishabituation. The same transgene rescues pharyngeal activity defects and both the habituation and dishabituation deficits seen in the eat-4 worms. Our results suggest that neurotransmitter regulation plays a role in habituation and may play a role in dishabituation. PMID- 10818170 TI - mu-Opioid receptors often colocalize with the substance P receptor (NK1) in the trigeminal dorsal horn. AB - Substance P (SP) is a peptide that is present in unmyelinated primary afferents to the dorsal horn and is released in response to painful or noxious stimuli. Opiates active at the mu-opiate receptor (MOR) produce antinociception, in part, through modulation of responses to SP. MOR ligands may either inhibit the release of SP or reduce the excitatory responses of second-order neurons to SP. We examined potential functional sites for interactions between SP and MOR with dual electron microscopic immmunocytochemical localization of the SP receptor (NK1) and MOR in rat trigeminal dorsal horn. We also examined the relationship between SP-containing profiles and NK1-bearing profiles. We found that 56% of SP immunoreactive terminals contact NK1 dendrites, whereas 34% of NK1-immunoreactive dendrites receive SP afferents. This result indicates that there is not a significant mismatch between sites of SP release and available NK1 receptors, although receptive neurons may contain receptors at sites distant from the peptide release site. With regard to opioid receptors, we found that many MOR immunoreactive dendrites also contain NK1 (32%), whereas a smaller proportion of NK1-immunoreactive dendrites contain MOR (17%). Few NK1 dendrites (2%) were contacted by MOR-immunoreactive afferents. These results provide the first direct evidence that MORs are on the same neurons as NK1 receptors, suggesting that MOR ligands directly modulate SP-induced nociceptive responses primarily at postsynaptic sites, rather than through inhibition of SP release from primary afferents. This colocalization of NK1 and MORs has significant implications for the development of pain therapies targeted at these nociceptive neurons. PMID- 10818171 TI - Teaching pediatric cardiology. PMID- 10818172 TI - Electrocardiography and 24-hour electrocardiographic ambulatory recording (Holter monitor) studies in children infected with human immunodeficiency virus type 1. The Pediatric Pulmonary and Cardiac Complications of Vertically Transmitted HIV-1 Infection Study Group. AB - Limited data are available on the electrocardiogram and ambulatory electrocardiogram recording (Holter) in children infected with the human immunodeficiency virus type 1 (HIV-1). The purpose of this study was to estimate the prevalence and cumulative incidence of rhythm and conduction abnormalities in HIV-1-infected children. Electrocardiograms and Holter monitoring studies were performed annually on 205 HIV-1-infected children enrolled after 28 days of life (group I), 93 HIV-1-infected infants enrolled during pregnancy or during the first 28 days of life (group IIa), and 463 HIV-1-uninfected infants enrolled during pregnancy or during the first 28 days of life (group IIb). The 5-year cumulative incidence in the group I children of second-degree atrioventricular block or supraventricular or ventricular tachycardia was 13.4%, and the 5-year incidence was higher for the older infected group I children (16.8% for children > or =4 years old at first study and 11.4% for children <4 years, p = 0.04). The mean corrected QT interval was also longer for the older infected group I children (p = 0.002) and prolonged in the HIV-1-infected compared to the HIV-1 uninfected group II children (p = 0.02). None of the children had atrial fibrillation or flutter. Arrhythmias are uncommon in children infected with HIV-1 and in children of HIV-1-infected mothers and the arrhythmias identified tend to be benign. Therefore, routine Holter monitoring does not appear to be indicated in asymptomatic children. PMID- 10818173 TI - Noninvasive prediction of pulmonary artery pressure in patients with isolated ventricular septal defect. AB - Management of patients with isolated ventricular septal defect (VSD) requires information regarding pulmonary artery pressure (PAP). The purpose of this study was to evaluate the individual predictive value of noninvasive methods for assessment of PAP and to determine if any combination of techniques significantly improved their predictive power. We reviewed the clinical history, electrocardiogram, and echocardiogram of 31 patients (age 1.9 +/- 1. 73 years) who underwent catheterization for isolated VSD. Noninvasive data were compared for patients with mean PAP <20 mmHg (group 1) and those with mean PAP > or =20 (group 2) at catheterization. Fourteen (45%) patients were in group 1 and 17 (55%) in group 2. Doppler estimation of VSD gradient, right ventricular hypertrophy by echocardiogram, interventricular septal orientation, and VSD size had predictive value for elevated mean PAP (p < 0.01). All patients (n = 6) with normal findings in all four variables had normal PAP. All patients (n = 12) with at least three of four abnormal findings had elevated PAP. Six patients in group 1 had at least one variable that incorrectly predicted high PAP, whereas 3 patients with normal findings on three of the four variables nevertheless had elevated PAP. No single noninvasive variable accurately predicted PAP in all cases. However, normal findings for all four significant variables did predict normal PAP and suggest that cardiac catheterization is unnecessary in that setting. However, any other combination of normal and abnormal findings for the four significant variables did not reliably predict PAP and such patients may require catheterization to directly measure PAP. PMID- 10818174 TI - The use of high-resolution computed tomography in the evaluation of pulmonary hemodynamics in patients with congenital heart disease: in pulmonary vessels larger than 1 mm in diameter. AB - High-resolution computed tomography (HRCT) was carried out in 36 patients with congenital left-to-right shunt disease and 10 normal control subjects to assess the feasibility of CT in the evaluation of pulmonary hemodynamics. The patients had a left-to-right or a bidirectional shunt and the hemodynamic data obtained by cardiac catheterization in these patients were compared to the information obtained by CT imaging. The pulmonary/systemic blood flow (Q(p)/Q(s)) ratio and pulmonic/systemic resistance (R(p)/R(s)) ratio had a significant correlation with the pulmonary artery/bronchus (PA/Br) ratio (r = 0.54 and r = -0.37, respectively) and pulmonary vein/bronchus (PV/Br) ratio (r = 0.66 and r = -0.66, respectively), and the R(p)/R(s) and mean PA pressure also showed a significant correlation with the PA/PV ratio (r = 0.53 and r = -0.61, respectively) in the mid-lung field when accompanying bronchi were 4. 0-5.9 mm in diameter. There was no correlation between the hemodynamic data and the size of the central and hilar PA or with the rate of PA tapering. With HRCT, it is possible to evaluate pulmonary hemodynamics in patients with congenital heart disease with a left-to right or bidirectional shunt, particularly R(p)/R(s) and mean PA pressure, which have been very difficult to obtain noninvasively. The small-sized pulmonary vessel/Br ratio or the small-sized PA/PV ratio could offer very useful information, but the dimension of the central PA provided the least useful information. PMID- 10818175 TI - Exercise capacity in young patients after total repair of Tetralogy of Fallot. AB - Tetralogy of Fallot is the most common form of cyanotic congenital heart disease. Measurement of physical activity is usually performed as a routine part of the patient's cardiac evaluation. The aim of this study was to examine the exercise performance of young patients operated on for tetralogy of Fallot, assessing the possible influence of known negative prognostic factors related to the surgical repair. The study group comprised 41 consecutive patients (29 male and 12 female, ages 11.2 +/- 3.9 years, range 6-16 years) operated on for tetralogy of Fallot. Patients in the study group were divided in subgroups in relation to the age of surgical intervention (before or after 2 years of life), the surgical approach (combined transatrial/transpulmonary approach or right ventriculotomy), and the presence of aortopulmonary shunts prior to performing total correction. Their data were compared with those of 33 aged-matched asymptomatic control subjects (19 male and 14 female, ages 11.9 +/- 1.3 years, range 11-16 years). Blood pressure and heart rate measured at rest were similar between control and Fallot groups. A normal increase in systolic blood pressure was observed in response to exercise intensity for all subgroups. No significant difference between control and Fallot groups was found under conditions of mild or moderate exercise or for diastolic blood pressure at rest and in response to exercise. Lower maximal heart rate and systolic blood pressure values were recorded in all patients when compared with the control subjects. Significant differences in peak workload were detected between control and Fallot groups and between the control and each subgroup; however, no difference was found between subgroups. In conclusion, despite their very satisfactory clinical status, all patients showed a reduced peak workload, irrespective of the surgical approach, age at surgery, and aortopulmonary shunts prior to performing total correction. PMID- 10818177 TI - A direct comparison of right and left ventricular performance in the isolated neonatal pig heart. AB - A comparison is presented between the performance of the right ventricle (RV) and the left ventricle (LV) in neonatal hearts studied under conditions of volume loading and tachycardia. Isolated, atrially paced (150 or 300 bpm), isovolumically beating pig hearts (1-3 days of age) underwent retrograde aortic perfusion with a nonrecirculating, crystalloid solution. Ventricular pressure was assessed with saline-filled balloon catheters, which allowed separate loading of the RV or LV. Both ventricles showed an initial increase followed by a leveling off, but no decline, in peak systolic pressure (PSP) and +dP/dt(max) with volume loading up to an end-diastolic pressure (EDP) of 18 mmHg. The LV generated a higher PSP and +dP/dt(max) compared to the RV at equivalent pressure or volume preloads. However, the maximal systolic elastance (E(max)) was comparable for both ventricles. Although the RV demonstrated a greater compliance than the LV, the myocardial relaxation time constant (tau) was similar for both chambers at equivalent volume preloads (sarcomere stretch). Positive dP/dt(max) correlated closely and in the same linear fashion with -dP/dt(max) for both ventricles, indicating that the RV and LV exhibited similar contraction-relaxation coupling. Increasing the heart rate to 300 bpm decreased PSP, +dP/dt(max), and -dP/dt(max) and increased EDP for both ventricles, whereas E(max) and tau were not significantly altered. Thus, although there are differences between the functional properties of the neonatal RV and LV, there are also important similarities, especially with regard to myocardial relaxation. PMID- 10818178 TI - Percutaneous pulmonary artery gene transfer in pigs using naked plasmid DNA delivered during angioplasty. AB - Gene transfer techniques are increasingly being used to study blood vessel biology and develop models for gene therapy. To date, there are no reports of pulmonary vascular gene transfer performed either without adjunctive agents or during angioplasty. We sought to demonstrate the feasibility of recombinant gene transfer to the pulmonary artery of juvenile pigs using naked plasmid DNA delivered via percutaneous angioplasty techniques. Plasmid DNA directing the expression of beta-galactosidase was used to transfect one pulmonary artery while the contralateral vessel served as an untreated control. One delivery technique used a standard angioplasty balloon coated with a DNA-heparin mixture. The second involved infusion of DNA between an angioplasty balloon and a surrounding, microporous balloon. Vessels were harvested 3 or 4 days following gene delivery. Protein expression was demonstrated by immunohistochemical staining in transfected but not control vessels in 9/9 pigs. Vascular wall expression was limited to endothelial cells. Pulmonary artery gene transfer using naked plasmid DNA delivered via percutaneous angioplasty techniques is feasible. Using naked plasmid DNA removes the potential for toxicity associated with adjunctive agents. The described techniques provide novel methods for studying pulmonary vascular biology in vivo and for developing future gene therapies. PMID- 10818180 TI - Correlation of plasma adrenomedullin to myocardial preservation during open-heart surgery. AB - Adrenomedullin (ADM) is a vasoactive peptide with potent dilatory effects. We studied whether perioperative myocardial injury could be altered by the presence of ADM. Blood samples from 19 children with congenital heart disease undergoing surgical repair were collected at six time points: preoperative, on cardiopulmonary bypass (CPB), and 0, 3, 6, and 12 hours after CPB. Blood levels of ADM (pg/ml) and troponin-I (Tn-I; ng/ml), a specific marker of myocardial injury, were measured. Patients were divided into three groups based on their 12 hour Tn-I levels (I, < 10, n = 6; II, 10-25, n = 6; III, >25, n = 7). Preoperative Tn-I levels were within the normal range for all patients. Preoperative ADM levels in group I (with little or no evidence of myocardial injury) were significantly greater than those of either group II or III (242.7 +/ 15.4 vs 83.8 +/- 18 and 85.2 +/- 5.5, respectively; p < or = 0.0001 for each). The 12-hour ADM levels in group I remained significantly lower than preoperative levels (242.7 +/- 15.4 vs 197.4 +/- 11.6, p < or = 0.03) but higher than in the other groups. In group III, ADM increased at the 12-hour time point (159.2 +/- 6.5, p < or = 0.0001 vs baseline). Higher preoperative ADM levels are associated with lower levels of myocardial injury (as assessed by troponin-I release) during congenital heart surgery. PMID- 10818181 TI - A comparison of fetal echocardiography in university and health maintenance organization settings. AB - Fetal echocardiographic findings, and decisions to continue or to terminate affected pregnancies, may differ between university (UNIV) and health maintenance organization (HMO) settings. The aim of this study was to review the fetal echocardiographic experience at a combined university/health maintenance organization program over a 4-year period. Imaging and counseling for affected pregnancies were provided by the same, single investigator at both sites. Out of a total of 1382 studies (940 UNIV, 442 HMO), 127 abnormals were identified (94 UNIV, 33 HMO). Among the 127 pregnancies with fetal heart disease, 24 (19%) underwent elective termination, 16% at UNIV and 27% at HMO (p = 0.2). Mean gestational age at the time of diagnosis was 25.2 weeks at UNIV compared with 22.3 weeks at HMO (p = 0.002). At UNIV, only 51% of diagnoses were made before 24 weeks compared with 79% at HMO (p = 0.003). Screening fetal sonograms, performed between 18 and 20 weeks on every pregnancy at HMO but not at UNIV, enabled earlier detection of congenital heart disease (CHD) and allowed more women with severely affected pregnancies the option to terminate. In both settings, indications with the highest yields for CHD included a right-sided stomach, abnormal four-chamber view, sustained bradycardia, abnormal fetal karyotype, fetal omphalocele, and maternal indomethacin. An echogenic reflector was identified in 86 pregnancies (7%) and did not represent a risk factor for CHD. No major differences in CHD were found between UNIV and HMO. In summary, this study found a significantly earlier diagnosis of CHD at HMO than at UNIV. This discrepancy between programs may explain, at least in part, the trend toward a higher frequency of decisions to terminate affected pregnancies at HMO than at UNIV, despite similar fetal findings. PMID- 10818183 TI - Radioangioscintigraphy and Doppler echocardiography in the quantification of left to-right shunt. AB - The magnitude of left-to-right shunt in 55 children with isolated congenital heart disease [atrial septal defect (ASD) or ventricular septal defect (VSD) (muscular and perimembranous)] was estimated by two methods: radionuclide quantification and Doppler echocardiography [flow (L/min) = mean velocity x area x ejection time x heart rate]. We found little difference between the magnitude of left-to-right shunt obtained with Doppler echocardiography and that with radioangioscintigraphy for a whole group of patients (N = 55, -11.42% to 12.04%) and for subgroups of ASD (n = 24, -12.49% to 12.19%) and VSD (n = 31, -10.69% to 12.23%). These results indicate that Doppler echocardiography, in comparison with radioangioscintigraphy, is sufficiently accurate for clinical estimation of the Q(p)/Q(s) ratio in isolated congenital heart disease with left-to-right shunt. PMID- 10818184 TI - Course of right and left ventricular function in patients with pulmonary insufficiency after repair of Tetralogy of Fallot. AB - Surgical repair of tetralogy of Fallot (TOF) frequently results in pulmonary valve insufficiency. Nevertheless, no serial information is available on the long term impact of the valvular insufficiency on right and left ventricular function. Right and left ventricular ejection fraction was measured serially by radionuclide angiocardiography in 21 patients with at least moderate pulmonary insufficiency after repair of TOF. A baseline study was obtained an average of 1.2 years after repair, and a follow-up study was performed an average of 10.2 years after surgery. Changes in ventricular function over time and deviations from the normal range were analyzed. At baseline evaluation the mean right ventricular ejection fraction (RVEF; 0.52 +/- 0.10) and left ventricular ejection fraction (LVEF; 0.68 +/- 0.10) were normal. At the time of follow-up the mean RVEF had significantly decreased to 0.45 +/- 0.09 (p < 0.01). The mean LVEF had decreased to 0.60 +/- 0.11 (p < 0.02). This change was independent of the RVEF (r = -0.13). Eleven patients (52%) had an abnormal RVEF or LVEF at follow-up. Nineteen patients (90%) showed a decrease of 0.05 or more in RVEF, LVEF, or both between studies. These data suggest a negative impact of long-standing pulmonary insufficiency on right and left ventricular systolic function after repair of TOF. Therefore, continued surveillance of biventricular function in this patient population appears warranted. PMID- 10818185 TI - Fetal echocardiography in ectopia cordis. AB - Ectopia cordis is an extremely rare congenital abnormality occurring in 5.5 to 7.9 per 1 million live births with high lethality. Between January 1995 and October 1997 eight cases of ectopia cordis were diagnosed at our institute before birth. On the basis of echocardiography the fetal heart anatomy was categorized as either normal heart anatomy (NHA; n = 3) or congenital heart defect (CHD; n = 5). In the majority of cases (seven of eight) other abnormalities were present. Some reports have described ectopia cordis being diagnosed in the first trimester of pregnancy. In our study group the average gestational age at diagnosis was 26 weeks. The prenatal diagnosis of isolated ectopia cordis is easy; counseling the patient, the perinatal management including term, place, and method of delivery, and optimal care of the newborn are more difficult. Ectopia cordis is a malformation that pediatricians rarely encounter, even at pediatric cardiology centers. Much more frequently it is a problem for sonographers and obstetricians; however, pediatric cardiologists should be aware of diagnostic algorithm for such cases, especially when additional abnormalities are present. PMID- 10818186 TI - Echocardiographic guidance for transcatheter coil embolization of congenital coronary arterial fistulas in children. AB - Congenital coronary arterial fistulas are rare anomalies that have traditionally been managed by surgical ligation. However, in recent years endovascular therapy has been employed with encouraging results. Between 1993 and 1996, we performed transcatheter coil embolization of coronary arterial fistulas to the right atrium or ventricle in four children ranging in age from 4.5 to 9.8 years. Cardiac and coronary arterial anatomy were diagnosed correctly on the preoperative echocardiogram in all patients, including the origin, course, and termination of the fistulas. The fistula was occluded completely in three of the patients, whereas trivial residual flow remained in the fourth. Transesophageal echocardiography was useful for monitoring the embolization procedure. In one of the patients, the fistula reopened while the child was on overnight heparin, although the magnitude of flow was less than that before the embolization. At follow-up ranging from 10 to 43 months, there was no flow through the fistula in any patient. We present our experience with these patients, with a focus on the importance of echocardiographic evaluation before, during, and after transcatheter therapy of coronary arterial fistulas. PMID- 10818187 TI - Unusual electrocardiogram findings in a preterm infant after fetal tachycardia with hydrops fetalis treated with flecainide. AB - Fetal tachycardia and signs of hydrops fetalis were diagnosed at 29 weeks of gestation. The heart rate normalized by combined treatment with digoxin and flecainide and was followed by improvement of the hydrops. Premature labor led to delivery at 33 weeks of gestation. The newborn infant showed mild respiratory distress and was in a hemodynamically stable condition. Marked QT segment anomalies on the electrocardiogram during the first postnatal days resolved completely within 3 weeks. They were unlikely to be attributable to myocarditis or myocardial infarction. We speculate that these anomalies were caused by the maternal flecainide therapy. PMID- 10818188 TI - Mycotic pseudoaneurysm of the aorta in children. AB - Mycotic pseudoaneurysm of the aorta is a rare disease in childhood. We report on two cases which were diagnosed in an unselected general pediatric population within an 8-month period. The first case was a 16-month-old toddler with a normal cardiac history who presented with purulent pericarditis due to group A streptococcus and subsequent pseudoaneurysm formation of the ascending aorta while convalescing from varicella infection. The second case was a 14-year-old girl with a previously undiagnosed coarctation of the aorta who developed a Staphylococcus aureus aortitis in the dilated poststenotic segment with pseudoaneurysm formation and infiltration into the adjacent lung tissue. In both cases parenteral antibiotic therapy was administered over 10 and 4 days, respectively, followed by emergency surgery consisting of aneurysmectomy, coarctectomy (case 2), and in situ homograft implantation. Recovery was uneventful. In both cases early institution of a femorofemoral cardiopulmonary bypass prevented a fatal outcome despite intraoperative rupture of the pseudoaneurysm. PMID- 10818189 TI - Heparin therapy and reversal of protein-losing enteropathy in a case with congenital heart disease. AB - We describe a patient with tricuspid hypoplasia who developed a protein-losing enteropathy. Having failed to respond to medical and surgical treatments, a heparin regimen was started with immediate decrease in enteric protein loss. PMID- 10818190 TI - Congenital subclavian artery to subclavian vein fistula presenting with congestive heart failure in an infant. AB - Contrary to acquired arteriovenous fistulas, the congenital malformations are very rare. Here we report a case of congenital arteriovenous fistula forming a communication between the right subclavian artery to subclavian vein. To our knowledge, there is no similar case in which an infant has been successfully operated on for cardiac failure. PMID- 10818192 TI - From PediHeart: inhaled nitric oxide-In clinical trial or in clinical practice? PMID- 10818191 TI - Transient neonatal cyanosis: unusual presentation of right-sided cardiac masses. AB - We report a case of a neonate with multiple cardiac masses, cyanosis, and a heart murmur. Arterial desaturation was the result of right-to-left shunting at the foramen ovale level caused by tricuspid regurgitation. Three right-sided cardiac masses were detected by echocardiography. By 2 weeks of age the patient had complete resolution of his cyanosis and improved tricuspid regurgitation following the normal decrease in pulmonary vascular resistance. At 2 years of age, he has no cardiovascular symptoms and the masses are calcified and have no hemodynamic consequences. PMID- 10818193 TI - Early presentation of a congenital coronary artery fistula in a neonate. AB - A neonate developed severe congestive heart failure secondary to a congenital coronary artery fistula requiring emergent surgery. Intraoperative transesophageal echocardiography helped guide successful emergent closure of the fistula without complications. PMID- 10818194 TI - From PediHeart: inhaled nitric oxide-references PMID- 10818195 TI - A quadricuspid pulmonic valve diagnosed in a live newborn by two-dimensional echocardiography. AB - Quadricuspid pulmonic valve is a rare congenital anomaly which appears to occur in the absence of other cardiac or systemic anomalies. It predominates in males and tends to be clinically quiescent. The first case of quadricuspid pulmonic valve in a live newborn infant diagnosed by two-dimensional echocardiography is presented here with a review of the literature. PMID- 10818196 TI - Atrioventricular block in a toxic child: do not forget diphtheria. AB - We describe a 4.5-year-old girl who presented with severe febrile throat infection and who, after a few days, developed ventricular tachycardia followed by atrioventricular block. Although a pacemaker was inserted, she died of cardiogenic shock. Throat cultures were positive for Corynebacterium diphtheriae. PMID- 10818197 TI - Atrial fibrillation associated with marijuana use. PMID- 10818198 TI - Selective coronary angiography in pediatric patients. AB - Selective coronary angiography (SCA) is an important diagnostic tool in pediatric cardiology; however, there are few reports on its feasibility and safety in young patients. We reviewed our experience with SCA from July 1, 1993 to December 31, 1997. There were 158 cardiac catheterizations that included SCA in patients whose ages ranged from 2 days to 46 years (median, 5.3 years). The most common indication was surveillance for coronary vasculopathy after heart transplantation. A retrograde approach was used in all patients through the femoral artery (n = 157) or umbilical artery (n = 1). Preformed coronary catheters were used and the Judkins left (JL) and Judkins right (JR) were the most common catheters, with the catheter curve size correlating with patient height (R(2) =.76 for JL, R(2) =. 673 for JR). Complications during SCA included brief ST-T wave changes (11%), bradycardia (2.5%), and ventricular fibrillation (0. 6%). Complications of vascular access were transient pulse loss (6%), hematoma (5%), and rebleeding (0.6%). Only one case of femoral artery occlusion was encountered on subsequent cath. In conclusion, complications of SCA were infrequent and serious complications were rare. SCA can be safely performed in pediatric patients at any age including neonates. PMID- 10818199 TI - An unusual variant of total anomalous pulmonary venous connection with varices and multiple drainage sites. AB - Total anomalous pulmonary venous connection (TAPVC) is an uncommon cardiac anomaly that has also rarely been associated with esophageal atresia. We report an unusual case of esophageal atresia with TAPVC with several varices and multiple drainage sites into the superior vena cava and portal vein. PMID- 10818201 TI - From other journals PMID- 10818200 TI - Protein-losing enteropathy after fontan operation: treatment with elementary diet in one case. PMID- 10818204 TI - Molecular cloning, gene expression, and identification of a splicing variant of the mouse parkin gene. AB - We have isolated mouse cDNA clones that are homologous to human Parkin gene, which was recently found to be responsible for the pathogenesis of autosomal recessive juvenile parkinsonism (AR-JP). One of these cDNA clones had the 1,392 bp open reading frame encoding a protein of 464 amino acids with presumed molecular weight of 51,615. The amino acid sequence of mouse parkin protein exhibits 83.2% identity to human Parkin protein, including the ubiquitin-like domain at the N-terminus (identity = 89.5%) and the RING finger-like domain at the C-terminus (identity = 90.6%). Two other clones had the 783-bp open reading frame encoding a truncated protein of 261 amino acids without RING finger-like domain. It was proved to be a novel splicing variant by 3'-RACE method. Northern blot analysis revealed that mouse parkin gene is expressed in various tissues including brain, heart, liver, skeletal muscle, kidney, and testis. It is notable that mouse parkin gene expression appears evident in 15th day mouse embryo and increases toward the later stage of development. These mouse parkin cDNA clones will be useful for elucidating the essential physiological function of parkin protein in mammals. PMID- 10818205 TI - A QTL on distal chromosome 3 that influences the severity of light-induced damage to mouse photoreceptors. AB - C57BL/6J-c(2J) (c2J) albino mice showed much less damage to their photoreceptors after exposure to prolonged light than BALB/c mice and seven other albino strains tested. There were no gender differences, and preliminary studies suggested that the c2J relative protective effect was a complex trait. A genome-wide scan using dinucleotide repeat markers was carried out for the analysis of 194 progeny of the backcross (c2J x BALB/c)F(1) x c2J and the thickness of the outer nuclear layer (ONL) of the retina was the quantitative trait reflecting retinal damage. Our results revealed a strong and highly significant quantitative trait locus (QTL) on mouse Chromosome (Chr) 3 that contributes almost 50% of the c2J protective effect, and three other very weak but significant QTLs on Chrs 9, 12, and 14. Interestingly, the Chrs 9 and 12 QTLs corresponded to relative susceptibility alleles in c2J (or relative protection alleles in BALB/c), the opposite of the relative protective effect of the QTLs on Chrs 3 and 14. We mapped the Rpe65 gene to the apex of the Chr 3 QTL (LOD score = 19.3). Northern analysis showed no difference in retinal expression of Rpe65 message between c2J and BALB/c mice. However, sequencing of the Rpe65 message revealed a single base change in codon 450, predicting a methionine in c2J and a leucine in BALB/c. When the retinas of aging BALB/c and c2J mice reared in normal cyclic light were compared, the BALB/c retinas showed a small but significant loss of photoreceptor cells, while the c2J retinas did not. Finding light damage-modifying genes in the mouse may open avenues of study for understanding age-related macular degeneration and other retinal degenerations, since light exposures may contribute to the course of these diseases. PMID- 10818206 TI - Isolation, genomic organization, and expression analysis of the mouse and rat homologs of MEFV, the gene for familial mediterranean fever. AB - Familial Mediterranean fever (FMF) is a recessive disorder characterized by episodes of fever with serositis or synovitis. Recently the FMF gene (MEFV) was cloned; the protein product, pyrin/marenostrin, is thought to regulate inflammation in myeloid cells. In this manuscript we report the mouse and rat homologs of MEFV. The murine gene contains ten exons with a coding sequence of 2304 bp, while the rat homolog has nine exons with a coding sequence of 2253 bp. A considerable amino acid sequence homology was observed between the mouse and human (47.6% identity and 65.5% similarity) and between the mouse and rat genes (73.5% identity and 82.1% similarity). The predicted rodent proteins have several important domains and signals found in human pyrin, including a B-box zinc finger domain, Robbins-Dingwall nuclear localization signal, and coiled-coil domain. However, perhaps because of an ancient frame-shift mutation, neither the mouse nor the rat protein has an intact C-terminal B30.2 domain, in which most FMF associated mutations have been found in human MEFV. Nevertheless, like the human gene, mouse Mefv is expressed in peripheral blood granulocytes but not lymphocytes. Consistent with its expression in granulocytes, Mefv was detected at high levels in the primary follicles and marginal zones of the splenic white pulp. Mefv is localized on mouse Chromosome (Chr) 16, region A3-B1, extending a region of synteny with human Chr 16p13.3. Development of knockout and knockin mouse models may provide further insights into the functional evolution of this gene. PMID- 10818207 TI - Gene encoding the mouse sulphamidase: cDNA cloning, structure, and chromosomal mapping. AB - Sulphamidase is an exoglycosidase involved in the degradation of heparan sulfate. Lack of sulphamidase activity leads to the lysosomal storage disorder Mucopolysaccharidosis type IIIA (Sanfilippo type A OMIM No. 252900). At present there are no naturally occurring small animal models of this disease that could be of fundamental importance to study the pathophysiology of the disease and to try therapeutic strategies. Cloning of the mouse gene is an important step to create a mouse model for this common mucopolysaccharidosis. We have isolated and sequenced the gene encoding mouse sulphamidase. Comparison of the deduced amino acid sequences of human and mouse sulphamidase showed 88% identity and 93% similarity. The exon-intron structure of the gene has been determined with the mouse 10-kb gene divided in 8 exons. The mouse sulphamidase gene (Sgsh) was mapped to the distal end of Chromosome (Chr) 11, in a region that is homologous with a segment of human Chr 17 containing the orthologous human gene. PMID- 10818208 TI - Gaping lids, gp, a mutation on centromeric chromosome 11 that causes defective eyelid development in mice. AB - In mammals, during fetal development, the eyelids grow and flatten over the eyes and temporarily fuse closed. Failure of this normal developmental process in mice leads to the defect, open-eyelids-at-birth. Nearly all newborns of the GP/Bc strain, homozygous for the spontaneous recessive mutation, gaping lids (gp), have bilateral open eyelids at birth, with essentially no fusion between the upper and lower eyelids. Histological sections and scanning electron microscopy of GP/Bc eyes during the normal period of eyelid growth and fusion indicate that gp/gp mutant fetuses have deficient upper and lower eyelids; surface periderm cells that appear to have some role in eyelid growth and fusion are present, but lack a normal "streaming pattern toward the fusion zone. No other defects due to the gaping lids mutation were detected. A genetic analysis based on outcrosses of GP/Bc to various linkage marker stocks and to CBA/J and ICR/Bc normal strains was done. Penetrance in F(2) segregants, but not in BC1 segregants, was usually significantly less than 100%, was strongly affected by the identity of the normal strain used, ranging from 44% to 92%, and indicated a potential complexity of modifiers. Forty-one affected F(2) and 120 BC(1) segregants from the outcross of GP/Bc to CBA/J, and 23 affected F(2) segregants from the outcross to ICR/Bc, were used to map gp to proximal Chr 11 between the centromere and D11Dal1 (Camk2b), an interval previously defined as less than 1 cM. Sets of whole F(2) litters from the crosses to CBA/J (n = 106) and ICR/Bc (n = 65) strains were typed for informative SSLPs near gp (D11Mit62 and D11Mit74, respectively) and demonstrated that the segregation ratios in the region are Mendelian. The known genes in the interval, Nf2 and Lif, do not seem to be obvious candidate genes for gp. An Egfr null allele was used to confirm the previously reported map position of the potential candidate locus, Egfr, to a more distal interval, between D11Mit62/226 and D11Mit151, from which gp had been excluded. Tests for allelism showed that the Egfr mutation and the gp mutation complement each other, and therefore also indicate that they are at different gene loci. Open-eyelids-at-birth is associated with several mutations at other loci with variable penetrance owing to modifiers and in other more complex genetic liabilities in inbred strains, and the genetics of this trait is a model for other genetically complex developmental threshold traits. The gaping lids mutation identifies a previously unknown locus on proximal Chromosome (Chr) 11 that has a strong role in fetal eyelid growth. PMID- 10818210 TI - Refined genetic and comparative physical mapping of the canine copper toxicosis locus. AB - Recently, the copper toxicosis (CT) locus in Bedlington terriers was assigned to canine chromosome region CFA10q26, which is homologous to human chromosome region HSA2p13-21. A comparative map between CFA10q21-26 and HSA2p13-21 was constructed by using genes already localized to HSA2p13-21. A high-resolution radiation map of CFA10q21-26 was constructed to facilitate positional cloning of the CT gene. For this map, seven Type I and eleven Type II markers were mapped. Using homozygosity mapping, the CT locus could be confined to a 42.3 cR(3000) region, between the FH2523 and C10.602 markers. On the basis of a partial BAC contig, it was estimated that 1-cR(3000) is equivalent to approximately 210 kb, implying that the CT candidate region is therefore estimated to be about 9 Mb. PMID- 10818209 TI - Isolation, characterization, expression and functional analysis of the zebrafish ortholog of MEN1. AB - Mutations in the MEN1 gene lead to an autosomal dominant disorder, multiple endocrine neoplasia type 1 (MEN1), which is characterized by tumors of the parathyroid, entero-pancreatic neuroendocrine, and pituitary tissues. The protein encoded by MEN1, 610-amino acid menin, resides primarily in the nucleus and binds to the transcription factor JunD, resulting in the repression of JunD-induced transcription. We report here a detailed characterization of the zebrafish men1 gene and its full-length (2551 nt) transcript, encoding a 617-amino acid protein with 67% identity and 80% similarity to human menin. Of the 81 missense mutations and in-frame deletions reported in MEN1 patients, 72 occur in residues that are identical in zebrafish, suggesting the importance of the conserved regions. The zebrafish men1 gene maps 61 cM from the top of linkage group 7 (LG7), a region that appears to show conserved synteny to the MEN1 loci at human 11q13. A 2.7-kb men1 message is detected at all stages of zebrafish development analyzed, from one-cell embryos to adult fish. Whole-mount in situ hybridization showed ubiquitous distribution of men1 message in zebrafish embryos at cleavage, blastula, gastrula, and early segmentation stages, with relatively abundant expression in blood cell progenitors (24 h post fertilization) and mesenchymal tissues (48 h post fertilization) at later stages. Zebrafish menin binds both human and mouse JunD, and represses JunD-induced transcription, indicating that the JunD-binding ability of menin is evolutionarily conserved. PMID- 10818211 TI - The upstream region of the mouse xist gene contains two ribosomal protein pseudogenes. PMID- 10818212 TI - A locus responsible for osteochondrodysplasia (ocd) is located on rat chromosome 11. PMID- 10818213 TI - Conserved linkage of NK-2 homeobox gene pairs Nkx2-2/2-4 and Nkx2-1/2-9 in mammals. PMID- 10818214 TI - Determination of the gene structure of human oligophrenin-1 and identification of three novel polymorphisms by screening of DNA from 164 patients with non-specific X-linked mental retardation. AB - We have recently shown that mutations in oligophrenin-1 (OPHN1) are responsible for non-specific X-linked mental retardation (MRX). The structure of the gene encoding the OPHN1 protein was determined by isolation of genomic DNA clones from the human cosmid library. Genomic fragments containing exons were sequenced, and the sequences of the exons and flanking introns were defined. Knowledge of the genomic structure of the OPHN1 gene, which spans at least 500 kb and consists of 25 exons, will facilitate the search for additional mutations in OPHN1. OPHN1 was screened for mutations in 164 subjects with non-specific mental retardation. Three nucleotide substitutions were identified, one of which was a silent mutation in the codon threonine 301 at position 903 (G-->C). The other substitutions were located in exon 2, a G-->A substitution at position 133 (A45T), and in exon 10, a C-->T substitution at position 902 (T301M), but these are common polymorphisms rather than disease-causing mutations. PMID- 10818215 TI - Evaluation of prenatal diagnosis of cleft lip/palate by foetal ultrasonographic examination. AB - Ultrasound scans in the midtrimester of pregnancy are now a routine part of antenatal care in many countries. This type of screening procedure can detect serious foetal anomalies. Thanks to our registry of congenital anomalies a study was undertaken. The objective of the study was to evaluate prenatal detection of cleft lip (palate)(CL/P) by routine ultrasonographic examination of the foetus in 265679 consecutive pregnancies from 1979 to 1998. The percentage of prenatal detection of CL/P was low. For isolated malformation (foetuses with only CL/P) the detection rate was low: 17.8%; however, this detection rate increased from 5.3% during the period 1979-1988 to 26.5% during the period 1989-1998, for foetuses with associated malformations (foetuses with CL/P and one or more additional major malformations) these detection rates were 34.6, 13. 3 and 50.0%, respectively. In foetuses with associated malformations with CL/P this detection rate was higher for chromosomal abnormalities with CL/P and for non-syndromic, non-chromosomal multiply malformed children with CL/P than for non-chromosomal recognized syndromes with CL/P. PMID- 10818216 TI - Formation of uniparental disomy 7 delineated from new cases and a UPD7 case after trisomy 7 rescue. Presentation of own results and review of the literature. AB - Maternal uniparental disomy for the entire chromosome 7 (matUPD7) has been reported several times in Silver-Russell syndrome (SRS) and growth-restricted patients. Here we present our results from the analysis of an abortion with confined placental mosaicism (CPM) for trisomy 7 which showed a maternal meiotic origin of the trisomy in the placenta and rescue to maternal UPD7 in foetal membrane. Furthermore, two newly detected SRS cases with maternal UPD7 revealed isodisomy and partial heterodisomy, respectively. Summarising these results with those published previously on the origin of UPD7, similar numbers of isodisomy (n=11) and cases with complete or partial heterodisomy (n=12) have been reported. In respect to the different formation mechanisms of UPD, complete isodisomy should be the result of a post-zygotic mitotic segregation error, whereas heterodisomic UPDs should be caused by trisomic rescue after meiotic non disjunction events. In maternal UPD7, 50% of cases seem to be caused by post zygotic mitotic segregation errors, which is similar to the situation in trisomy 7. This result corresponds to the situation in trisomy 8 but is in contrast to observations in the frequent aneuploidies. Thus, the different findings in these aberrations reflect the presence of multiple factors that act to ensure normal segregation, varying in importance for each chromosome. PMID- 10818217 TI - Origin of trisomy 21 in Down syndrome cases from a Spanish population registry. AB - We have carried out a population-based study on the origin of the extra chromosome 21 in 38 families with Down syndrome (DS) offspring in El Valles (Spain). From 1991 to 1994, a higher prevalence of DS (22.7/10000 live births, stillbirths and induced abortions) was found compared to the majority of EUROCAT registries. The distribution of trisomy 21 by origin was 88% maternal (90.6% meiosis I, 6.2% meiosis II, 3.1% maternal mosaicism), 5.6% paternal (50% meiosis I, 50% meiosis II) and 5.6% mitotic. The percentage of parental mosaicism was 2.7%. These percentages are similar to those previously reported. Recombination study revealed a maternal meiosis I genetic map of 32.68 cM (approximately one half the length of the normal female map). Mean maternal age among non recombinant cases involving MI errors was significantly lower (31.1 years) than among those cases showing one observable crossover (36.1 years) (P<0.05); this could support the hypothesis that 'achiasmate' chromosomes may be subject to aberrant segregation regardless of maternal age. PMID- 10818218 TI - Molecular diagnosis of the fragile X and FRAXE syndromes in patients with mental retardation of unknown cause in Mexico. AB - The fragile X syndrome (Fra-X) is the most common cause of inherited mental retardation with X-linked semi-dominant inheritance. The prevalence of Fra-X in the Mexican population is unknown. The aim of this population screening study was to determine if Fra-X or FRAXE mutations are the cause of a number of cases of mental retardation in a sample of Mexican children with mental retardation of unknown cause (MRUC) and to stress the importance of performing molecular analysis of the FMR-1 gene in all patients with MRUC. We report here the direct analysis of CGG and GCC repeats within the FMR-1 and FMR-2 genes, respectively, in 62 unrelated patients with MRUC. Two male index cases had the CGG expansion, although they did not express the Xq27.3 fragile site cytogenetically. Fra-X diagnosis was highly suspected on a clinical basis in one of the patients, but not in the other. Both mothers were found to be premutation carriers. The molecular studies of FMR-1 showed that the proportion of MRUC patients with Fra-X is 3.2%. This frequency was not significantly different to that reported in most populations. As reported in other series, no patients with FRAXE were found in our sample. Our findings confirm that the molecular analysis of the FMR-1 gene is necessary in MRUC patients to achieve unequivocal diagnosis of fragile X syndrome, carrier premutation detection and for accurate genetic counseling. PMID- 10818219 TI - Cerebellar ataxia and mental retardation in a child with an inherited satellited chromosome 4q. AB - We identified a case of familial satellited chromosome 4q through a child with cerebellar ataxia and mental retardation. No loss of genetic material could be demonstrated at the molecular level but other possible mechanisms of this association are discussed. We conclude that in these cases genetic counselling should be reassuring. PMID- 10818220 TI - Hypomelanosis of ito and a 'mirror image' whole chromosome duplication resulting in trisomy 14 mosaicism. AB - We describe a female infant with multiple congenital anomalies including unusual hyperpigmentation, tetralogy of Fallot, absent corpus callosum and wide prominent nasal bridge. The infant was initially seen for genetic consultation on day one after birth. Chromosome analysis from cultured lymphocytes showed a normal 46,XX karyotype. However, cultured skin fibroblasts showed mosaicism with 46,XX,add(14)(q32).ish psu dic dup(14)(q32p13)(wcp14+)/46,XX complements. A review of the published report with chromosome mosaicism and hypomelanosis of Ito (HMI) is included. We suggest that the trisomy 14 mosaicism seen in fibroblast cultures has importance in the expression of pigmentation dysplasias in this patient. Pigmentary anomaly may be due to loss or gain of specific genes that influence pigmentation located on the long arm of chromosome 14 in this patient. PMID- 10818221 TI - Molecular cytogenetics, RFLP analysis and clinical characterization of a de novo trisomy 10p case. AB - A new case of a de novo trisomy 10cen-->10pter is described. The karyotype was exactly defined by high resolution banding and FISH analysis; the chromosome aberration was of maternal meiotic origin as demonstrated by RFLP analysis. Clinical data are reported and correlated with other trisomy 10p cases from the literature. A critical review of the literature was made to define the phenotype of trisomy 10p syndrome. PMID- 10818222 TI - inv(9)(p24q13) in three sterile brothers. AB - Only nine non-polymorphic constitutional pericentric inversions of chromosome 9 have been described. We report on a familial inv(9)(p24q13) associated with sterility in three brothers. The mother's chromosomes were normal in blood lymphocytes (n=130); the father was already deceased and his karyotype unknown. However, the presence of any of the maternal chromosomes 9 (as assessed by C banding) in her carrier children is inconsistent with the assumption of maternal mosaicism. Two single sisters were also carriers. The same rearranged chromosome 9 in the three sterile brothers can hardly be regarded as a fortuitous association, especially when the breakpoints are almost identical to those of the sole inversion previously found in an azoospermic male. If their father was a carrier, the observed sterility may be the result of 'chromosome anticipation', a phenomenon already invoked for certain familial chromosomal rearrangements. PMID- 10818223 TI - An intracranial carcinoma in a Mexican woman with Bloom syndrome. AB - An intracranial cell squamous carcinoma was found in a 27-year-old Mexican woman with Bloom syndrome (BS), including growth retardation, sun-sensitive telangiectatic erythema, defective immunity, and increased number of mitotic chiasmata and sister chromatid exchanges. The tumour, probably originating from the inner or middle ear epithelium, was resected but the patient died a few days after surgery. There was no parental consanguinity nor Jewish or European ancestry for at least five generations; in fact, her parents were mostly indigenous people. This case represents an undescribed intracranial malignancy in BS and the third Mexican BS patient reported. The typical BS phenotype in a woman with pigmented skin challenges the contention that pigmented females are less severely affected. PMID- 10818224 TI - alpha-Lactalbumin: structure and function. AB - Small milk protein alpha-lactalbumin (alpha-LA), a component of lactose synthase, is a simple model Ca(2+) binding protein, which does not belong to the EF-hand proteins, and a classical example of molten globule state. It has a strong Ca(2+) binding site, which binds Mg(2+), Mn(2+), Na(+), and K(+), and several distinct Zn(2+) binding sites. The binding of cations to the Ca(2+) site increases protein stability against action of heat and various denaturing agents, while the binding of Zn(2+) to the Ca(2+)-loaded protein decreases its stability. Functioning of alpha-LA requires its interactions with membranes, proteins, peptides and low molecular weight substrates and products. It was shown that these interactions are modulated by the binding of metal cations. Recently it was found that some folding variants of alpha-LA demonstrate bactericidal activity and some of them cause apoptosis of tumor cells. PMID- 10818225 TI - The in vitro activity of ADAM-10 is inhibited by TIMP-1 and TIMP-3. AB - A recombinant soluble form of the catalytic domain of human ADAM-10 was expressed as an Fc fusion protein from myeloma cells. The ADAM-10 was catalytically active, cleaving myelin basic protein and peptides based on the previously described 'metallosheddase' cleavage sites of tumour necrosis factor alpha, CD40 ligand and amyloid precursor protein. The myelin basic protein degradation assay was used to demonstrate that hydroxamate inhibitors of matrix metalloproteinases (MMPs) were also inhibitors of ADAM-10. The natural MMP inhibitors, TIMP-2 and TIMP-4 were unable to inhibit ADAM-10, but TIMP-1 and TIMP-3 were inhibitory. Using a quenched fluorescent substrate assay and ADAM-10 we obtained approximate apparent inhibition constants of 0.1 nM (TIMP-1) and 0.9 nM (TIMP-3). The TIMP-1 inhibition of ADAM-10 could therefore prove useful in distinguishing its activity from that of TACE, which is only inhibited by TIMP-3, in cell based assays. PMID- 10818226 TI - alpha-Actinin is a potent regulator of G protein-coupled receptor kinase activity and substrate specificity in vitro. AB - G protein-coupled receptor kinases (GRKs) phosphorylate G protein-coupled receptors, thereby terminating receptor signaling. Herein we report that alpha actinin potently inhibits all GRK family members. In addition, calcium-bound calmodulin and phosphatidylinositol 4,5-bisphosphate (PIP2), two regulators of GRK activity, coordinate with alpha-actinin to modulate substrate specificity of the GRKs. In the presence of calmodulin and alpha-actinin, GRK5 phosphorylates soluble, but not membrane-incorporated substrates. In contrast, in the presence of PIP2 and alpha-actinin, GRK5 phosphorylates membrane-incorporated, but not soluble substrates. Thus, modulation of alpha-actinin-mediated inhibition of GRKs by PIP2 and calmodulin has profound effects on both GRK activity and substrate specificity. PMID- 10818227 TI - The RIP-like kinase, RIP3, induces apoptosis and NF-kappaB nuclear translocation and localizes to mitochondria. AB - A RIP-like protein, RIP3, has recently been reported that contains an N-terminal kinase domain and a novel C-terminal domain that promotes apoptosis. These experiments further characterize RIP3-mediated apoptosis and NF-kappaB activation. Northern blots indicate that rip3 mRNA displays a restricted pattern of expression including regions of the adult central nervous system. The rip3 gene was localized by fluorescent in situ hybridization to human chromosome 14q11.2, a region frequently altered in several types of neoplasia. RIP3-mediated apoptosis was inhibited by Bcl-2, Bcl-x(L), dominant-negative FADD, as well as the general caspase inhibitor Z-VAD. Further dissection of caspase involvement in RIP3-induced apoptosis indicated inhibition by the more specific inhibitors Z DEVD (caspase-3, -6, -7, -8, and -10) and Z-VDVAD (caspase-2). However, caspase 1, -6, -8 and -9 inhibitors had little or no effect on RIP3-mediated apoptosis. Mutational analysis of RIP3 revealed that the C-terminus of RIP3 contributed to its apoptotic activity. This region is similar, but distinct, to the death domain found in many pro-apoptotic receptors and adapter proteins, including FAS, FADD, TNFR1, and RIP. Furthermore, point mutations of RIP3 at amino acids conserved among death domains, abrogated its apoptotic activity. RIP3 was localized by immunofluorescence to the mitochondrion and may play a key role in the mitochondrial disruptions often associated with apoptosis. PMID- 10818228 TI - Subcellular differences in post-translational modification of barley 14-3-3 proteins. AB - Expression and post-translational modification of barley 14-3-3 isoforms, 14-3 3A, 14-3-3B and 14-3-3C, were investigated using isoform-specific antibodies. Although all three isoforms were shown to be present in the cytosolic, the nuclear and the microsomal cell fractions, differences in post-translational modification were identified for the different cell fractions. Germination related modifications of 14-3-3 proteins were observed in the cytosol and the microsomal fraction, but not in the nucleus. In vitro proteolytic cleavage of 14 3-3 proteins using trypsin suggests that for 14-3-3A this change was caused by proteolytic cleavage of the unconserved C-terminal region. PMID- 10818229 TI - Apical and non-polarized secretion of serpins from MDCK cells. AB - Corticosteroid binding globulin, a member of the serpin family, was previously shown to be secreted mainly apically from MDCK cells in an N-glycan independent manner [Larsen et al. (1999) FEBS Lett. 451, 19-22]. Apart from N-glycosylation, serpins are not known to carry any other posttranslational modifications, suggesting the presence of a proteinaceous apical sorting signal. In the present study we have expressed four other members of the serpin family: alpha1 antitrypsin, C1 inhibitor, plasminogen activator inhibitor-1 and antithrombin in MDCK cells. Tight monolayers of transfected cells were grown on filters and the amounts of recombinantly expressed serpins in the apical and the basolateral media were determined. alpha1-Antitrypsin and C1 inhibitor were found mainly in the apical medium whereas plasminogen activator inhibitor-1 and antithrombin were found in roughly equal amounts in the apical and basolateral media. Control experiments showed that all four serpins are transported along the exocytotic pathway in an uncomplicated way that does not involve transcytosis or differences in stability on the two sides of the cells. We conclude that some members of the serpin family including corticosteroid binding globulin, alpha1-antitrypsin and C1 inhibitor are secreted mainly apically from MDCK cells whereas plasminogen activator inhibitor-1 and antithrombin are secreted in a non-polarized manner. PMID- 10818230 TI - Molecular cloning, characterization and evolution of the gene encoding a new group of short-chain alpha-neurotoxins in an Australian elapid, Pseudonaja textilis. AB - The structure and organization of five genes responsible for the synthesis of six isoforms of short-chain alpha-neurotoxins in Pseudonaja textilis venom are reported in this paper. This also forms the first report which describes the synthesis of two neurotoxin mRNA variants from one of these genes (Pt-sntx1) as a result of alternative splicing. Each gene consists of three exons which are separated by two introns and each has a functional promoter. The promoter activity was confirmed by both CAT assay and Real-Time PCR. A transcription initiation site, two putative TATA boxes, one CCAAT box and the transcription factor binding consensus sites for AP-1, GATA-2, c/EBPb were identified in the 5' non-coding region of each gene. Phylogenetic analysis showed that these five genes from P. textilis constituted a distinct group which has evolved by gene duplication followed by accelerated evolution from an ancestral gene. PMID- 10818231 TI - Galectin-3 overexpression protects from cell damage and death by influencing mitochondrial homeostasis. AB - Galectins are a family of proteins involved in several cell processes, including their survival and death. Galectin-3 has in particular been described as an anti apoptotic molecule entangled with a number of subcellular activities including anoikis resistance. In this work we partially address the mechanisms underlying this activity pointing at two key factors in injury progression: the alteration of mitochondrial membrane potential and the formation of reactive oxygen species. Overexpression of galectin-3 appears in fact to exert a protective effect towards both these events. On the basis of these data, we propose a reappraisal of the role of galectin-3 as a regulator of mitochondrial homeostasis. PMID- 10818232 TI - Sequence, genomic structure and tissue expression of carp (Cyprinus carpio L.) vertebrate ancient (VA) opsin. AB - We report the isolation and characterisation of a novel opsin cDNA from the retina and pineal of the common carp (Cyprinus carpio L.). When a comparison of the amino acid sequences of salmon vertebrate ancient opsin (sVA) and the novel carp opsin are made, and the carboxyl terminus is omitted, the level of identity between these two opsins is 81% and represents the second example of the VA opsin family. We have therefore termed this C. carpio opsin as carp VA opsin (cVA opsin). We show that members of the VA opsin family may exist in two variants or isoforms based upon the length of the carboxyl terminus and propose that the mechanism of production of the short VA opsin isoform is alternative splicing of intron 4 of the VA opsin gene. The VA opsin gene consists of five exons, with intron 2 significantly shifted in a 3' direction relative to the corresponding intron in rod and cone opsins. The position (or lack) of intron 2 appears to be a diagnostic feature which separates the image forming rod and cone opsin families from the more recently discovered non-visual opsin families (pin-opsins (P), vertebrate ancient (VA), parapinopsin (PP)). Finally, we suggest that lamprey P opsin should be reassigned to the VA opsin family based upon its level of amino acid identity, genomic structure with respect to the position of intron 2 and nucleotide phylogeny. PMID- 10818233 TI - Molecular characterization of a calmodulin-like dictyostelium protein CalB. AB - A gene named calB was cloned and characterized in Dictyostelium. A relationship to calmodulin (CaM) is suggested by sequence identity (50%), similar exon-intron structure and cross-reactivity with anti-CaM sera. The level of calB mRNA is developmentally regulated with maxima during aggregation and in spores. CalB null cells grow normally, develop and produce viable spores. We demonstrated the capacity of tagged CalB to bind Ca(2+) using the (45)Ca(2+) overlay assay and showed that its mobility on SDS-PAGE is dependent on Ca(2+)/EGTA pretreatment. PMID- 10818234 TI - Genetic evidence of branching in the isoprenoid pathway for the production of isopentenyl diphosphate and dimethylallyl diphosphate in Escherichia coli. AB - An alternative mevalonate-independent pathway for isoprenoid biosynthesis has been recently discovered in eubacteria (including Escherichia coli) and plant plastids, although it is not fully elucidated yet. In this work, E. coli cells were engineered to utilize exogenously provided mevalonate and used to demonstrate by a genetic approach that branching of the endogenous pathway results in separate synthesis of the isoprenoid building units isopentenyl diphosphate (IPP) and its isomer dimethylallyl diphosphate (DMAPP). In addition, the IPP isomerase encoded by the idi gene was shown to be functional in vivo and to represent the only possibility for interconverting IPP and DMAPP in this bacterium. PMID- 10818235 TI - Activation of PPARdelta alters lipid metabolism in db/db mice. AB - Peroxisome proliferator-activated receptors (PPARs) are nuclear receptors, which heterodimerize with the retinoid X receptor and bind to peroxisome proliferator response elements in the promoters of regulated genes. Despite the wealth of information available on the function of PPARalpha and PPARgamma, relatively little is known about the most widely expressed PPAR subtype, PPARdelta. Here we show that treatment of insulin resistant db/db mice with the PPARdelta agonist L 165041, at doses that had no effect on either glucose or triglycerides, raised total plasma cholesterol concentrations. The increased cholesterol was primarily associated with high density lipoprotein (HDL) particles, as shown by fast protein liquid chromatography analysis. These data were corroborated by the chemical analysis of the lipoproteins isolated by ultracentrifugation, demonstrating that treatment with L-165041 produced an increase in circulating HDL without major changes in very low or low density lipoproteins. White adipose tissue lipoprotein lipase activity was reduced following treatment with the PPARdelta ligand, but was increased by a PPARgamma agonist. These data suggest both that PPARdelta is involved in the regulation of cholesterol metabolism in db/db mice and that PPARdelta ligands could potentially have therapeutic value. PMID- 10818236 TI - Isopentenyl diphosphate isomerase deficiency in Synechocystis sp. strain PCC6803. AB - Isopentenyl diphosphate isomerase (IPP isomerase) in many organisms and in plastids is central to isoprenoid synthesis and involves the conversion between IPP and dimethylallyl diphosphate (DMAPP). It is shown that Synechocystis PCC6803 is deficient in IPP isomerase activity, consistent with the absence in its genome of an obvious homologue for the enzyme. Incorporation of [1-(14)C]IPP in cell extracts, primarily into C(20), occurs only upon priming with DMAPP in Synechocystis PCC6803 and in Synechococcus PCC7942. Isoprenoid synthesis in these cyanobacteria does not appear to involve interconversion of IPP and DMAPP, raising the possibility that they are not within the plastid evolutionary lineage. PMID- 10818237 TI - Nature of linkage between the cationic headgroup and cholesteryl skeleton controls gene transfection efficiency. AB - Three novel cationic cholesterol derivatives with different modes of linkage between the cationic headgroup and the cholesteryl backbone have been synthesized and used as mixtures with 1, 2-dioleoyl-L-alpha-glycero-3-phosphatidyl ethanolamine (DOPE) for liposome-mediated gene transfection. A pronounced improvement in gene transfer efficiency was observed when the cationic center was appended to the cholesteryl backbone using an ether linkage as opposed to when the linkages were based on either ester or urethane groups. Amphiphiles with ether links such as cholest-5-en-3beta-oxyethane-N,N,N-trimethyl ammonium bromide (2) and cholest-5-en-3beta-oxyethane-N,N-dimethyl-N-2-hydroxyethyl ammonium bromide (3) showed transfection efficiencies considerably greater than commercially available gene transfer reagents. Notably, the transfection ability of 2 with DOPE in the presence of serum was significantly greater than Lipofectamine((R)) and Lipofectin((R)). Interestingly, 3 did not require the helper lipid DOPE for transfection. This suggests that these newly described cholesterol-based amphiphiles should be very promising in liposome-mediated gene transfection. The advantage that the ether linkage possesses would be important in the design of newer, more efficient cholesterol-based delivery reagents. PMID- 10818238 TI - Switch of histamine receptor expression from H2 to H1 during differentiation of monocytes into macrophages. AB - It is known that histamine suppresses gene expression and synthesis of tumor necrosis factor alpha (TNF-alpha) induced by lipopolysaccharide (LPS) in human peripheral blood mononuclear monocytes (HPM) or alveolar macrophages via histamine H2 receptors. We investigated the effect of histamine and differentiation in macrophages on the expression and secretion of TNF-alpha, TNF alpha-converting enzyme (TACE), and histamine H1 and H2 receptors by use of a leukemia cell line, U937, and HPM. Differentiation of U937 and HPM cells with 12 O-tetradecanoylphorbol-13-acetate (TPA) enhanced the H1 receptor expression and rather suppressed the H2 receptor, resulting in up-regulation of the histamine induced expression and secretion of TNF-alpha, modulated via TACE. Therefore, histamine failed to inhibit up-regulated expression of TNF-alpha induced by LPS in macrophages. The switch from H2 to H1 receptors during differentiation in the monocyte/macrophage lineage could participate in the pathogenic processes of atherosclerosis and inflammatory reactions in the arterial wall. PMID- 10818239 TI - Pairing of oligosaccharides in the Fc region of immunoglobulin G. AB - The Fc portion of immunoglobulin G (IgG) expresses paired oligosaccharides with microheterogeneities, which are associated with efficiencies of effector functions and with pathological states. A comparison of electrospray ionization mass spectrometry data obtained using a variety of Fc fragments derived from human and mouse IgG that do and do not retain the inter-chain disulfide bridge(s) revealed that (1) the Fc portion can be asymmetric as well as symmetric with respect to glycosylation and (2) the ratios of the individual glycoforms are different from what is expected from the random pairing. PMID- 10818240 TI - Analysis of post-translational modification of mycobacterial proteins using a cassette expression system. AB - A recombinant expression system was developed to analyse sequence determinants involved in O-glycosylation of proteins in mycobacteria. By expressing peptide sequences corresponding to known glycosylation sites within a chimeric lipoprotein construct, amino acids flanking modified threonine residues were found to have an important influence on glycosylation. The expression system was used to screen mycobacterial sequences selected using a neural network (NetOglyc) trained on eukaryotic O-glycoproteins. Evidence of glycosylation was obtained for eight of 11 proteins tested. The results suggest that sites involved in O glycosylation of mycobacterial and eukaryotic proteins share similar structural features. PMID- 10818241 TI - A novel putative transcription factor protein MYT2 that preferentially binds supercoiled DNA and induces DNA synthesis in quiescent cells. AB - Myelin transcription factor 2 (MYT2), a putative transcription factor found in the human central nervous system, was cloned from an expression cDNA library from human T-cells. MYT2 shares weak similarity to bacterial type I topoisomerases and shares 63% sequence identity to a replicase from Leuconostoc mesenteroides. MYT2 preferentially binds supercoiled DNA (scDNA). Incubation of MYT2 and scDNA at or above equal molar ratios generated topoisomer-like patterns that were abolished by deproteination. Thus, MYT2 appears to relax scDNA via a non-enzymatic mechanism. The banding pattern of MYT2-scDNA complexes was shown to be quantisized, saturable and sequence-independent. Microinjection of MYT2 mRNA induced G(o) growth-arrested NIH 3T3 cells to enter the S phase of the cell cycle. PMID- 10818242 TI - Multiple glandular origins of queen pheromones in the fire ant Solenopsis invicta. AB - The poison sac of the fire ant Solenopsis invicta is the only identified glandular source of pheromones produced by a functional ant queen. This structure, which contains the poison gland, has previously been shown to be the source of a releaser pheromone that mediates queen recognition and tending by workers. The poison sac has also been demonstrated to be the source of a primer pheromone that inhibits winged, virgin queens from shedding their wings (dealating) and developing their ovaries. To determine if the poison sac was the only source of these pheromones, we excised the poison sac from queens and observed whether operated queens retained their pheromonal effects. In a first experiment, the poison sac was removed from functional (egg-laying) queens which were then paired with unoperated nestmate queens in small colonies. Counts of the workers surrounding each queen two weeks after the operation showed that queens without poison sac were as effective as their unoperated nestmates in attracting worker retinues. In a second experiment, we removed the poison sacs of virgin queens which had not yet begun laying eggs and thus had not begun producing queen pheromone. After allowing them to develop their ovaries, these individuals produced amounts of queen recognition pheromone comparable to those secreted by unoperated or sham operated virgin queens as determined by bioassay. Testing the head, thorax and abdomens of functional queens separately revealed that the head was the most attractive region in relation to its relative surface area. Bioassays of extracts of two cephalic glands-the mandibular gland and postpharyngeal gland-showed that the postpharyngeal gland is a second source of the queen recognition pheromone. Finally, we found that virgin queens whose poison sacs were removed before they began producing queen pheromone initiated production of a primer pheromone that inhibits winged virgin queens from dealating, indicating that this pheromonal effect also has an additional but as yet undetermined source. These results parallel those on the honey bee in which several of the pheromonal effects of functional queens appear to have multiple glandular sources. PMID- 10818243 TI - Racing against host's immunity defenses: a likely strategy for passive evasion of encapsulation in Asobara tabida parasitoids. AB - The hymenopteran Asobara tabida Nees (Braconidae, Alysiinae) develops as a solitary endophagous parasite in larvae of several Drosophila species. Most A. tabida eggs possess a sticky chorion which attaches to the tissue of the host organs within a few hours following oviposition. A. tabida sticky eggs usually avoid encapsulation, though the probability of survival decreases in hosts carrying a larger number of circulating hemocytes. Here, we hypothesized that the elicitation of the encapsulation reaction may result from a race between two phenomena: the host's hemocytic reaction and the embedment of the parasitic egg within the host tissues. In order to test this hypothesis, we measured the speed of capsule formation in D. melanogaster larvae of different ages, knowing that the number of circulating hemocytes increases with the age of the larvae. Using a non-virulent A. tabida strain, the eggs of which do not attach to the host tissue, we found that the speed of capsule formation increased correlatively with the age of the D. melanogaster larva. Therefore, the hypothesis of a physiological race between host's immunity defenses and parasite's avoidance of host's defenses is strongly supported by our results. Also, A. tabida eggs which attach to the host's tissue before the attack by the hemocytes has taken place may be considered as a strategy of passive evasion from encapsulation. PMID- 10818244 TI - The role of AB - The effect of [His(7)]-corazonin on the body color in Locusta migratoria was examined by varying the injected dose and the time of injection in both an albino and a normal (pigmented) strain. Albino nymphs injected with a high dose (100pmol) of [His(7)]-corazonin at the beginning of the third instar turned completely black in the following instar, whereas those injected with the same dose in the middle of the instar developed black patterns with an orange background color, the body coloration characteristic of normal gregarious (crowded) individuals. Injection at the end of the third instar induced a reddish color with few black spots. Irrespective of the time of injection of the peptide, most of these individuals became completely black after ecdysis to the fifth instar. A similar result was obtained with a lower dose (1pmol), although the color expressed was lighter. In the normal strain, injection of 1nmol or 100pmol into crowded third instar nymphs also caused most of them to become completely black in the fourth and fifth instars, but a lower dose apparently had no influence. These results suggest that the temporal changes in hemolymph titer of [His(7)]-corazonin are important in the expression of body color in L. migratoria. PMID- 10818245 TI - Salivary proteins of aphids, a pilot study on identification, separation and immunolocalisation. AB - Salivary proteins (SPs) of Schizaphis graminum, Acyrthosiphon pisum and Myzus persicae were studied after probing and feeding on different artificial diets. Salivary sheaths as well as apical lumps of saliva were found, presumably representing subsequently excreted saliva of different types. Phenoloxidase, pectinase and peroxidase activities were detected by staining the enzyme converted products, thus confirming these enzyme activities found earlier by others. Proteinase and cellulase were not found. SPs in three major SDS-PAGE bands, at 154 and 66/69 kDa, were collected in fluid diets (soluble fraction) and as sheath material (solid fraction) attached to the membranes covering these diets. Proteins of both fractions presumably represented the enzymatic activities found, although this could not be proven. The lack of electrophoretic mobility of the undenaturated (isoelectrofocusing and PAGE) active proteins meant that they could not be separated, whereas the mobile denaturated (SDS-PAGE) proteins had lost their enzyme activity. Polyclonal antibodies, anti-SP154 and anti-SP66/69, both cross-reacted to most salivary proteins in Western blots. They also reacted to sheath material and to the principal salivary glands. For further studies of saliva some monoclonal antibodies were developed. The complexity of salivation and the relation of the results obtained to the behaviourally known secretion periods is discussed. PMID- 10818246 TI - Thermoregulation of water collecting honey bees (Apis mellifera). AB - Honey bees (Apis mellifera carnica, Apidae, Hymenoptera) visited a pond in order to collect water. During their stays at the pond the body surface temperature of water foragers was measured using contactless thermography. Irrespective of the ambient temperature (T(A)) which ranged from 13.6 to 27.2 degrees C, the water carriers reached thoracic temperatures of 36-38.8 degrees C (mean values of the measuring periods). The maximum thoracic value of an individual bee was 44.5 degrees C. At higher T(A) (20.9-27.2 degrees C) head and abdomen were only about 3 degrees C and 2 degrees C on the average higher than the surroundings, respectively. In the lower range of T(A) (13.6-16.6 degrees C), however, the bees warmed their heads up to 29.2 degrees C (13 degrees C above T(A)) and the abdomen up to 23.3 degrees C (7.1 degrees C above T(A); mean values of the measuring periods).The head and abdomen were even provided independently of one another with heat from the thorax. At a higher T(A) only little heat came from the heated thorax into the abdomen, at a cooler T(A) (13.6-16.6 degrees C) more heat reached the abdomen. In all probability, at a higher T(A) only a small amount of haemolymph was pumped from the thorax into the abdomen; the most warm blood probably circulated in the head-thorax area. The average duration of stays at the pond decreased linearly from 110 to 42 s with rising T(A). Head and thorax showed great fluctuations of temperature. For example, the head was heated by 4.6 degrees C within 25 s, the thorax by 6.1 degrees C within 30 s.Foragers drinking sucrose solution are known to increase their thoracic temperature with rising concentration of the sucrose solution. The water foragers had thoracic temperatures similar to that of bees feeding on 0.5 molar sucrose solution. It is hypothesized that the foraging motivation of both groups was similar and therefore they regulated their thoraces at the same temperature level. PMID- 10818247 TI - Central processing of sex pheromone stimuli is differentially regulated by juvenile hormone in a male moth. AB - In the male moth, Agrotis ipsilon, the neuronal basis for juvenile hormone (JH) linked modulation of sex pheromone responsiveness was investigated following stimulation of the antenna with i) an extract of female pheromone gland, ii) the synthetic pheromone blends from A. ipsilon and a closely related species, A. segetum, and iii) single components of the A. ipsilon blend. Response characteristics of olfactory interneurons were studied in the antennal lobe (AL) at different ages and with manipulated JH levels using intracellular recording techniques. Blend-specific, generalist and component-specific neurons were identified and described according to their response pattern. The proportion of low threshold AL interneurons increased significantly with age for all stimuli tested. Changes were, however, less pronounced for the minor single components. The proportion of low threshold AL interneurons in allatectomized (JH-deprived) mature males was significantly lower for all stimuli than in intact mature males. A large proportion of low threshold AL interneurons responding to the pheromone blends, but not as pronounced for single pheromone components, could be restored/induced by injecting JH either into JH-deprived mature males or into young immature males. The specificity for the species-specific blend compared to the A. segetum blend increased with age and JH injections. PMID- 10818248 TI - Nutrient absorption and utilization by wing and flight muscle morphs of the cricket Gryllus firmus: implications for the trade-off between flight capability and early reproduction. AB - Absorption efficiency (AD, approximate digestibility, assimilation efficiency) of various macronutrients and conversion of absorbed nutrients to biomass (ECD) were compared among the two types of flightless morph and the flight-capable morph of the cricket, Gryllus firmus. No biologically significant phenotypic or genetic difference in AD for carbohydrate, protein or lipid was observed among morphs fed either a high-nutrient (100%) or a low-nutrient (25%) diet. Thus, previously documented differences among adult morphs in carbohydrate and lipid content must be caused by processes other than variation in nutrient absorption by morphs during adulthood. Relative absorption efficiency of total dry mass of food by morphs of G. firmus appears to be a valid indicator of relative AD of total calories. Morphs did not differ phenotypically or genetically in the excretion of end products of nitrogen metabolism (uric acid, hypoxanthine plus xanthine) on either the high nutrient or the low nutrient diet. Nutritional indices corrected for excreted nitrogenous metabolites were very similar to uncorrected indices, and the pattern of variation among the morphs was the same for corrected or uncorrected values. Each of the two types of flightless morph converted a greater proportion of absorbed nutrients into body mass, mainly ovaries, and allocated a smaller proportion of assimilated nutrients to respiration than did the flight capable morph. Moreover, the trade-off between respiration and early reproduction was substantially magnified on the low nutrient diet. These results extend previous findings of a trade-off between flight capability and early reproduction in wing-polymorphic Gryllus species (1) to diets of very different nutrient quantity, and (2) to flightlessness arising from different causes: blockage of flight muscle development in juveniles vs histolysis of fully-developed flight muscles in adults. PMID- 10818249 TI - Terpenoid omega-hydroxylase (CYP4C7) messenger RNA levels in the corpora allata: a marker for ovarian control of juvenile hormone synthesis in Diploptera punctata. AB - Ribonuclease protection assays were used to measure changes in allatal transcript levels of the CYP4C7 gene which encodes a cytochrome P450 terpenoid omega hydroxylase thought to play a role in the metabolism of JH and its precursors. Denervation of the corpora allata does not affect the pattern of expression of the CYP4C7 gene. Transplantation experiments show that CYP4C7 mRNA levels are dependent on a humoral factor characteristic of the reproductive state of the insect. Messenger RNA levels rise substantially in mated or denervated females, or in mated or virgin females treated with hydroprene, when the follicle length is over 1.5 mm. Vitellogenic ovaries however exert a negative influence on CYP4C7 expression, as ovariectomy in mated females causes a premature rise in CYP4C7 mRNA levels. The half-life of the CYP4C7 transcript is approx. 2 h when the corpora allata are incubated in vitro. Under these conditions, coincubation with a post-vitellogenic ovary maintains high CYP4C7 transcript levels in the glands. Excess juvenile hormone or analog applied at the end of vitellogenesis blocks ovulation or causes abortion of embryos deposited in the brood sac. We conclude that expression of the CYP4C7 gene is tightly controlled by the ovary, and it coincides with the ovarian signal to turn off juvenile hormone synthesis. The role of the CYP4C7 enzyme may be to ensure the clearance of allatal juvenile hormone and its precursors at the end of the gonotrophic cycle. PMID- 10818250 TI - Native and heterologous neuropeptides are cardioactive in Drosophila melanogaster. AB - Nine neuropeptides isolated from Drosophila melanogaster and five neuropeptides, previously isolated from the CNS of Limulus with antisera to FMRFamide-related peptides, were tested for their effects on the myogenic heart of Drosophila melanogaster. Of the native peptides, TDVDHVFLRF-NH(2) (Dromyosuppressin), DPKQDFMRFamide, and PDNFMRFamide significantly slowed the heart. Of the Limulus peptides, DEGHKMLYFamide (LP1) increased heart rate significantly, GHSLLHFamide (LP2) and PDHHMMYFamide (LP3) decreased the heart's rate, while DHGNMLYFamide (LP4) and GGRSPSLRLRFamide (LP5) had no effect at the concentrations we employed. Dromyosuppressin, DPKQDFMRFamide, and PDNFMRFamide from Drosophila, and LP2 and LP3 from Limulus, which belong to a novel group of peptides structurally unrelated to FMRFamide, are among only a very few substances from within the general group of neuropeptides and neurohormones known to slow the heart of Drosophila, and as such offer an important tool for investigating the molecular mechanisms underlying the control of the pacemaker. PMID- 10818251 TI - Neuropeptides--an overview. AB - The present article provides a brief overview of various aspects on neuropeptides, emphasizing their multitude and their wide distribution in both the peripheral and central nervous system. Interestingly, neuropeptides are also expressed in various types of glial cells under normal and experimental conditions. The recent identification of, often multiple, receptor subtypes for each peptide, as well as the development of peptide antagonists, have provided an experimental framework to explore functional roles of neuropeptides. A characteristic of neuropeptides is the plasticity in their expression, reflecting the fact that release has to be compensated by de novo synthesis at the cell body level. In several systems peptides can be expressed at very low levels normally but are upregulated in response to, for example, nerve injury. The fact that neuropeptides virtually always coexist with one or more classic transmitters suggests that they are involved in modulatory processes and probably in many other types of functions, for example exerting trophic effects. Recent studies employing transgene technology have provided some information on their functional role, although compensatory mechanisms in all probability could disguise even a well defined action. It has been recognized that both 'old' and newly discovered peptides may be involved in the regulation of food intake. Recently the first disease-related mutation in a peptidergic system has been identified, and clinical efficacy of a substance P antagonist for treatment of depression has been reported. Taken together it seems that peptides may play a role particularly when the nervous system is stressed, challenged or afflicted by disease, and that peptidergic systems may, therefore, be targets for novel therapeutic strategies. PMID- 10818252 TI - Effects of the CRF(1) receptor antagonist, CP 154,526, in the separation-induced vocalization anxiolytic test in rat pups. AB - CRF(1) receptor antagonists have been proposed as novel pharmacological treatments for depression, anxiety and stress disorders. The primary goal of the present study was to evaluate the effects of the CRF(1) receptor antagonist, CP 154,526, in the separation-induced vocalization (SIV) model of anxiety. Nine- to 11-day-old rat pups were separated from their litter and the effects of intraperitoneally administered test compounds on the elicited ultrasonic vocalizations were measured. Side-effect potential was assessed using a modified inclined plane test ('time on an inclined plane', or TIP), and using negative geotaxis. SIV was reduced by CP 154,526 at doses that did not affect TIP or negative geotaxis, a profile like that of the 5-HT(1A) partial agonist buspirone. The benzodiazepine anxiolytic, diazepam, decreased SIV but also produced significant side effects at one to three-fold higher doses. Additional pharmacological characterization of SIV demonstrated anxiolytic-like effects of the atypical antipsychotic, clozapine, but not the typical antipsychotic, haloperidol, and of the serotonin reuptake inhibitor, zimelidine, but not the norepinephrine reuptake inhibitor, desipramine. In summary, the data support the conclusion that selective CRF(1) receptor antagonists may have utility in anxiety and stress disorders. The data further support the use of separation-induced vocalizations for identifying mechanistically diverse compounds with anxiolytic actions in man. PMID- 10818253 TI - Evidence for the abundant expression of arginine 185 containing human CRF(2alpha) receptors and the role of position 185 for receptor-ligand selectivity. AB - The abundance of a histidine residue at position 185 (His(185)) of the human corticotropin-releasing factor (CRF) type 2 alpha receptor (hCRF(2alpha)) was investigated. His(185) has only been reported in hCRF(2); CRF(2) proteins from other species and all CRF(1) receptors encode an arginine (Arg(185)) at the corresponding position. Cloning of partial and full-length hCRF(2) cDNAs from a variety of neuronal and peripheral tissues revealed the existence of receptor molecules encoding Arg(185) only. Sequence analysis of the hCRF(2) gene verified the existence of Arg(185) also on genomic level. Full-length cDNAs encoding either the His(185) (R2H(185)) or the Arg(185) (R2R(185)) variants of hCRF(2alpha) were stably expressed in HEK293 cells and tested for ligand binding properties. In displacement studies R2H(185) and R2R(185) displayed a similar substrate specificity, human and rat urocortin, and the peptide antagonists astressin and alpha-helical CRF((9-41)) were bound with high affinity whereas human and ovine CRF were low-affinity ligands. Significant differences were observed for sauvagine and urotensin I, which bound with 3-fold (sauvagine) and 9 fold (urotensin I) higher affinity to R2R(185). These data indicate that hCRF(2), like all vertebrate CRF(1) and CRF(2) proteins encodes an arginine residue at the junction between extracellular domain 2 and transmembrane domain 3 and that this amino acid plays a role for the discrimination of some CRF peptide ligands. PMID- 10818254 TI - Galanin/alpha2-receptor interactions in central cardiovascular control. AB - The modulation of the central cardiovascular effects of alpha(2)-adrenoceptor activation by galanin and its N-terminal fragment galanin-(1-15) has been evaluated by quantitative receptor autoradiography and cardiovascular analysis. Intracisternal coinjections of threshold doses of galanin and the selective and hypotensive alpha(2)-receptor agonist clonidine induced rapid and maintained vasopressor and tachycardic responses (p<0.001) instead of a hypotensive response, whereas the coinjections of threshold doses of the N-terminal galanin fragment (1-15) and clonidine did not elicit significant cardiovascular changes. Receptor autoradiographical experiments showed that galanin (1 nM) significantly increased the K(d) (p<0.01) and B(max) values (p<0.01) of [(3)H]p-Aminoclonidine binding sites in the nucleus tractus solitarii (NTS) compatible with a possible antagonistic interaction with the alpha(2)-adrenoceptors, and this effect was blocked by the presence of the specific galanin receptor antagonist M35. In addition, clonidine (30 nM) induced a 50% increase in the B(0) values of galanin based on competition experiments with [(125)I]-galanin binding in the NTS. These findings suggest the existence of an antagonistic effect of galanin, but not of galanin fragment (1-15), on the cardiovascular responses mediated by alpha(2) receptors as well as a reciprocal facilitatory effect of alpha(2)-receptors on galanin binding. These mechanisms could be mediated by a reciprocal galanin alpha(2) receptor interaction within the NTS. PMID- 10818255 TI - Galanin and spatial learning in the rat. Evidence for a differential role for galanin in subregions of the hippocampal formation. AB - Anatomical, neurochemical and behavioural evidence support a role for galanin in hippocampally mediated functions such as spatial learning and memory. To obtain more precise information on this role, galanin (3 nmol/rat) was infused via bilateral chronic cannulae into different areas of the hippocampal formation which are characterized by different galanin receptor subtypes and also by different galanin innervation patterns. The effects of infused galanin on spatial learning were examined in the Morris swim maze. Infusions of galanin into both the dorsal and ventral dentate gyrus, which mainly contain GAL-R2 receptor mRNA and a high degree of galanin-noradrenaline coexistence, significantly retarded spatial acquisition without affecting swim speed or performance in the visible platform test. This spatial learning deficit was fully blocked by pretreatment with the non-selective galanin antagonist M35. Analysis of retention performance suggested that the major effect of intrahippocampal galanin is mediated via a specific disruption of acquisition mechanisms of importance for performance in the probe trial. Galanin infused into the ventral CA1 (a mainly GAL-R1 receptor mRNA expressing region) or into anterior, ventral CA3 regions did not produce any deficits in spatial learning compared to control animals. These results suggest that galanin mediates its action on spatial learning mainly through the GAL-R2 receptor subtype in areas where most of the galanin is present in noradrenergic terminals. A possible role for the GAL-R1 receptor subtype in cognition in the dorsal and ventral hippocampus remains to be defined. The results suggest a differential functional role for galanin and galanin receptor subtypes within subregions of the hippocampal formation. PMID- 10818256 TI - Galanin receptor plasticity within the nucleus basalis in early and late Alzheimer's disease: an in vitro autoradiographic analysis. AB - Hypertrophy of fibers containing galanin (GAL), the inhibitory neurotransmitter of acetylcholine, occur on remaining cholinergic nucleus basalis neurons in late stage Alzheimer's disease (AD). The present investigation evaluated whether changes in the number of GAL receptors (GALR) were detectable within the nucleus basalis in the early or late stage of AD when compared to age-matched controls. Postmortem neuropathological specimens were obtained at autopsy from three groups: late AD, early (possible) AD, and normal (age-matched controls) human subjects. Autoradiography of GALR binding was performed on human brain sections from each of the three groups. Analysis of autoradiographic images show no change in the distribution of ([125])hGAL binding sites in early AD cases throughout the nucleus basalis. In contrast, the number of ([125])hGAL binding sites was increased over the anterior nucleus basalis subfield in late stage AD. A region of-interest densitometric analysis of the anterior nucleus basalis in the late stage AD cases depict an increase in the number of ([125])hGAL binding sites by approximately two-three-fold when compared to normal (age-matched controls). Quantitative measures of ([125])hGAL binding densities were not significantly different in the anterolateral, intermediate or posterior nucleus basalis subsectors of early or late stage AD when compared to age-matched controls. These observations show that the occurrence of overexpression of GALRs coincide with earlier reports showing galaninergic fibers hyperinnervating surviving cholinergic basal forebrain neurons in late stage AD. PMID- 10818258 TI - Molecular determinants of peptide and nonpeptide NK-2 receptor antagonists binding sites of the human tachykinin NK-2 receptor by site-directed mutagenesis. AB - A series of 14 mutants on nine selected residues of the human tachykinin NK(2) receptor was produced and stably transfected into CHO cells to investigate the binding of the peptide MEN 11420 and the nonpeptide SR 48968 antagonists. The main interactions found for MEN 11420 were with Thr171, Tyr206, Tyr266 and Phe270. In the case of SR 48968 crucial residues were Tyr266 and Tyr289. While some overlapping of the binding sites exists, the binding modes suggested by this study appear not to allow structural correlation, and therefore general SAR, between these two antagonists. PMID- 10818257 TI - Pharmacological blockade or genetic deletion of substance P (NK(1)) receptors attenuates neonatal vocalisation in guinea-pigs and mice. AB - The regulation of stress-induced vocalisations by central NK(1) receptors was investigated using pharmacological antagonists in guinea-pigs, a species with human-like NK(1) receptors, and transgenic NK1R-/- mice. In guinea-pigs, i.c.v. infusion of the selective substance P agonist GR73632 (0.1 nmol) elicited a pronounced vocalisation response that was blocked enantioselectively by the NK(1) receptor antagonists CP-99,994 and L-733,060 (0.1-10 mg/kg). GR73632-induced vocalisations were also markedly attenuated by the antidepressant drugs imipramine and fluoxetine (30 mg/kg), but not by the benzodiazepine anxiolytic diazepam (3 mg/kg) or the 5-HT(1A) agonist buspirone (10 mg/kg). Similarly, vocalisations in guinea-pig pups separated from their mothers were blocked enantioselectively by the highly brain-penetrant NK(1) receptor antagonists L 733,060 and GR205171 (ID(50) 3 mg/kg), but not by the poorly brain-penetrant compounds LY303870 and CGP49823 (30 mg/kg). Separation-induced vocalisations were also blocked by the anxiolytic drugs diazepam, chlordiazepoxide and buspirone (ID(50) 0.5-1 mg/kg), and by the antidepressant drugs phenelzine, imipramine, fluoxetine and venlafaxine (ID(50) 3-8 mg/kg). In normal mouse pups, GR205171 attenuated neonatal vocalisations when administered at a high dose (30 mg/kg) only, consistent with its lower affinity for the rat than the guinea-pig NK(1) receptor. Ultrasound calls in NK1R-/- mouse pups were markedly reduced compared with those in WT pups, confirming the specific involvement of NK(1) receptors in the regulation of vocalisation. These observations suggest that centrally-acting NK(1) receptor antagonists may have clinical utility in the treatment of a range of anxiety and mood disorders. PMID- 10818259 TI - Distribution of the neurotensin receptor NTS1 in the rat CNS studied using an amino-terminal directed antibody. AB - The distribution of neurotensin receptor 1 immunoreactivity in the rat brain was studied using an antibody against the amino-terminal of the receptor expressed as a fusion protein with glutathione-S transferase. Affinity purified antibodies detected the fusion protein and the complete neurotensin receptor sequence expressed in Escherichia coli. The immunostaining was abolished by preabsorption with the amino-terminal fusion protein. Immunoreactive neurotensin receptor 1 immunoreactivity was detected on cell bodies and their processes in a number of CNS regions. In agreement with previous binding studies neurotensin receptor 1 immunoreactivity was particularly localised in cell bodies in the basal forebrain, nucleus basalis and substantia nigra. At the electron microscope level immunoreactivity was found both in axonal bouton and dendrites and spines in the basal forebrain indicating that neurotensin may act both pre- and post synaptically. There were several regions such as the substantia gelatinosa, ventral caudate-putamen and the lateral reticular nucleus where the neurotensin receptor 1 positive cells had not previously been reported, indicating that distribution of this receptor is widespread. PMID- 10818260 TI - Selective somatostatin sst(2) receptor blockade with the novel cyclic octapeptide, CYN-154806. AB - The cyclic octapeptide, CYN-154806, inhibited specific [(125)I]-[Tyr(11)]-SRIF binding to CHO-K1 cell membranes expressing human recombinant somatostatin (SRIF) sst(2) receptors (pIC(50) 8. 58) or rat sst(2(a)) and rat sst(2(b)) receptors (pIC(50) 8.35 and 8. 10, respectively). The affinity of CYN-154806 at other human somatostatin receptor types was at least 100 times lower (pIC(50) 5. 41-6.48). In functional studies, CYN-154806 inhibited SRIF-induced increases in extracellular acidification (EAR) in CHO-K1 cells expressing h sst(2) receptors (pK(B) 7.92) but had no effect on UTP-induced increases in EAR. CYN-154806 also blocked SRIF induced increases [(35)S]-GTPgammaS binding in CHO-K1 cell membranes expressing h sst(2) receptors as well as rat sst(2(a)) and rat sst(2(b)) receptors (pK(B) 7.81, 7.68 and 7.96, respectively). In marked contrast, no blockade was observed at h sst(5) receptors in concentrations as high 10 microM. The antagonistic activity of CYN-154806 was also studied in isolated tissue preparations that are known to express endogenous SRIF receptors. Thus CYN-154806 blocked SRIF, but not DAMGO-induced inhibition of neurogenic contractions in rat isolated vas deferens and guinea-pig ileum (pK(B) 7.79 and 7.49, respectively). CYN-154806 had no effect on SRIF-28 induced inhibition of neurogenic contractions in guinea-pig vas deferens. The results demonstrate that CYN-154806 is a highly potent specific and selective SRIF sst(2) receptor blocking drug. Furthermore, sst(2) receptors mediate SRIF-induced inhibition of neurogenic contractions in rat vas deferens and guinea-pig ileum but not guinea-pig vas deferens which is thought to be mediated by sst(5) receptors. PMID- 10818261 TI - Cloning, expression and pharmacological characterisation of the mouse somatostatin sst(5) receptor. AB - The mouse somatostatin (somatotropin release inhibiting factor, SRIF) sst(5) receptor coding sequence was cloned from a mouse BALB/c genomic library. It shows 97% and 81% homology with the corresponding rat and human receptors, respectively. The msst(5) receptor messenger RNA (mRNA) is present at low levels in the adult mouse brain, with significant expression in a few nuclei only, e.g. in the septum (lateral septal nuclei) or the amygdala (medial amygdaloid nucleus); very few signals were observed in the mesencephalon, metencephalon, and myelencephalon (except the dorsal motor nucleus of the vagus nerve). The msst(5) receptor was stably expressed in the hamster fibroblast cell line CCL39-SRE-Luci, which harbours the luciferase reporter gene driven by the serum responsive element. [(125)I]LTT-SRIF-28 ([Leu(8), D-Trp(22), (125)I-Tyr(25)]-SRIF-28), [(125)I]Tyr(10)-CST, [(125)I]CGP 23996, and [(125)I]Tyr(3)-octreotide labelled msst(5) receptors with high affinity (pK(d) values: 11.0, 10.15, 9.75 and 9.43) and in a saturable manner, but defined different Bmax values: 697, 495, 540 and 144 fmoles/mg, respectively. [(125)I]LTT-SRIF-28-labelled sites displayed the following rank order: SRIF-28> rCST-14> somatuline > CGP-23996= SRIF-14= octreotide, whereas [(125)I]Tyr(3)-octreotide-labelled sites displayed a different profile: octreotide > SRIF-28> rCST-14= somatuline > SRIF-14> CGP 23996. The pharmacological profiles determined with [(125)I]LTT-SRIF-28, [(125)I]CGP 23996 and [(125)I]Tyr(10)-CST correlated highly significantly (r(2) =0.88-0.99), whereas [(125)I]Tyr(3)-octreotide binding was rather divergent (r(2) =0.77). Also, human and mouse sst(5) receptor profiles are very different, e. g. r(2) =0.385 for [(125)I]Tyr(10)-CST and r(2) =0.323 for [(125)I]LTT-SRIF-28 labelled sites. Somatostatin induces expression of luciferase reporter gene in CCL39-SRE-Luci cells. The profile was consistent with a msst(5) receptor-mediated effect although apparent potency in the luciferase assay was much reduced compared to radioligand binding data: Octreotide = SRIF-28> rCST-14= SRIF-14= CGP 23996. Octreotide, SRIF-28, BIM23052 and D Tyr Cyanamid 154806 behaved as full or nearly full agonists in comparison to SRIF-14, whereas the other compounds had relative efficacies of 40 to 70%. The present study shows that agonists radioligands define apparently different receptor populations in terms of number of sites and pharmacological profile in cells expressing a single recombinant receptor. These variations suggest that the conformation of the ligand receptor complex may vary depending on the agonist. Further, the msst(5) receptor, although primarily coupled to Gi/Go proteins, is able to stimulate luciferase gene expression driven by the serum responsive element. Finally, it is suggested that putative sst(2) selective agonists e.g. octreotide, RC160 or BIM23027 show similar or higher potency at msst(5) receptors than SRIF-14. PMID- 10818262 TI - Involvement of hippocampal neuropeptide Y in mediating the chronic actions of lithium, electroconvulsive stimulation and citalopram. AB - Neuropeptide Y (NPY) has been considered to be involved in the pathogenesis of affective disorders, and chronic treatment with lithium or electroconvulsive stimuli (ECS) has been shown to increase mRNA and peptide levels of NPY in rat brain tissue. Consequently, parameters reflective of NPYergic neurotransmission were studied in the hippocampus of rats following chronic treatment with lithium, ECS or the selective serotonin re-uptake inhibitor (SSRI), citalopram. Lithium (28 days, diet) and ECS (10 days, once daily) treatments caused a marked increase in levels of preproNPY mRNA in the CA1 area and dentate gyrus (DG). This increase was accompanied by an increase in extracellular levels of NPY in the dorsal hippocampus of freely moving rats as determined by microdialysis, suggesting that lithium and ECS treatments lead to an increased biosynthesis and release of NPY in this area. (125)I-peptide YY (PYY) binding was reduced by 40 and 60% respectively in the DG following the same treatments, showing that the increased release is accompanied by a down-regulation of corresponding binding sites. In contrast, citalopram (10 mg/kg i.p., twice daily for 28 days) caused a 100% increase in (125)I-PYY binding in CA, CA3 and DG while levels of preproNPY mRNA and extracellular NPY in the hippocampus were unaffected. The results indicate that various agents and stimuli exerting antidepressant effects in humans, such as chronic lithium, ECS and citalopram all increase NPYergic neurotransmission in the hippocampus by distinct modes of action. Moreover, NPY (6 microg) given intracerebroventricularly (i.c.v.) induced an antidepressant-like effect in the forced swim test. It is hypothesised that the increase in NPYergic neurotransmission may be associated with the mechanism of action of various antidepressant treatments in the alleviation of depression. PMID- 10818263 TI - Neuropeptide Y promotes sleep and inhibits ACTH and cortisol release in young men. AB - Anxiolytic and sedative effects of neuropeptide Y (NPY) are thought to involve inhibition of corticotropin-releasing hormone (CRH). Enhanced secretion of CRH plays a critical role in the pathophysiology of major depression, characterized by sleep disturbances, anxiety and loss of appetite. We examined for the first time in young men effects of intravenous injections of NPY (4x50 or 100 microg, n = 9 and 11, respectively, at 22.00, 23.00, 24. 00 and 01.00 compared to saline) on the sleep electroencephalogram (EEG; recorded from 23.00 to 07.00) and nocturnal secretion of adrenocorticotrophic hormone (ACTH), cortisol, growth hormone (GH), prolactin and leptin. Repeated measures MANOVA showed that ACTH secretion during the first half of the night was reduced by the lower dose of NPY only (F = 8.7, p<0.05), while cortisol secretion during the second half of the night was reduced regardless of the dose (F = 7.9, p<0.05). Regardless of the dose, NPY enhanced sleep period time and stage 2 sleep (F = 12.8 and 5.4, each p<0.05), and also reduced sleep latency and time awake (F = 4.9 and 4.4, each p<0.05) and modulated REM sleep. In summary, NPY promotes sleep and inhibits the hypothalamo-pituitary-adrenocortical (HPA) axis in humans, pointing to a possible role of NPY agonists for the development of novel treatment strategies for affective disorders. PMID- 10818264 TI - Organic compounds in the extrapalial fluid and haemolymph of Anodonta cygnea (L.) with emphasis on the seasonal biomineralization process. AB - Bivalve mollusks, such as the freshwater mussel Anodonta cygnea, show seasonal changes in calcification. This cycle of calcification must either be a cause or a consequence for seasonal fluctuations in the organic composition of the animal's fluids, haemolymph and extrapallial fluid, the liquid media for biomineralization. We monitored the fluids of A. cygnea, throughout a 1-year cycle, for the presence of organic constituents, known to be important for biomineralization, such as proteins, glycosaminoglycans (GAGs) and hexosamines. Proteins were subjected to further study, namely through the total amino acid determination and fraction separation by agarose gel electrophoresis. GAG levels were fairly constant throughout the year, with a maximum concentration in July and a minimum in January, a feature also detected for glucosamine, although with higher fluctuations. Proteins showed highly increased concentrations during June and July, both in total amounts and individual fractions. All fractions showed similar trends throughout the year, with lowest general levels in October, the starting month of a period when some fractions were not detectable at all. All fractions ended this low period in May, when a sometimes-important increase could be detected. As to the total amino acid composition of the fluids, the general trend followed that of proteins, except for ornithine (Orn), a non-proteic amino acid. The overall fluctuations detected in the biological fluids of A. cygnea suggest that the main variation related to the calcification cycle must be quantitative, since no different compounds appear in specific periods, to achieve also specific results. PMID- 10818265 TI - Differential expression of ras in organs and embryos of shrimp Penaeus monodon (Crustacea: Decapoda). AB - Total RNA from shrimp hepatopancreas of Penaeus monodon showed three prominent bands that react with the shrimp ras probe, a 239-bp product, of approximately 4.8 kb (R1), 3.1 kb (R2) and 1.3 kb (R3) on the northern blot. The R1 is the least abundant. Analyses of total RNA from gill and heart were similar to each other. The highest expression of Ras was observed in the gill, while a negligible signal was detected with the Ras probe in muscle. Ras expression is developmentally regulated in embryonic stages of shrimp. Messenger RNA levels of ras were increased from a minimum in the nauplius stage to a maximum in the post larvae stage for R1 and R2. R3 showed a maximum at the protozoea stage. On the other hand, the activity of protein geranylgeranyltransferase I was increased significantly in the early nauplius stage. No correlative increase of prenylation activity by protein geranylgeranyltransferase I was observed with the transcription activity of ras. PMID- 10818266 TI - Effects of soybean meal and salinity on intestinal transport of nutrients in Atlantic salmon (Salmo salar L.) and rainbow trout (Oncorhynchus mykiss). AB - Groups of fresh- and seawater-adapted Atlantic salmon (Salmo salar L.) and rainbow trout (Oncorhynchus mykiss) were fed diets with (SBM diet) or without (control diet) extracted soybean meal (30% of protein substituted with SBM) for 3 weeks. Average fish size per group ranged from 597 to 1763 g. One tank or net pen per species, dietary group and water salinity was used. In vitro nutrient transport (D-glucose, the L-amino acids aspartate, lysine, methionine, phenylalanine and proline, and the dipeptide glycyl-sarcosine) was measured using intact tissue (everted sleeve method) from the different postgastric intestinal regions. The dimensions of the different intestinal regions were also measured for each treatment group. Results indicate that SBM causes decreased carrier mediated transport and increased permeability of distal intestinal epithelium for the nutrients, and the capacity of this region to absorb nutrient was diminished. Salinity may also affect the relative contribution of carrier-mediated and independent uptake to total nutrient absorption. PMID- 10818267 TI - Binding sites for human interferon-gamma in protocerebrum and hemolymph of tobacco hornworm (Manduca sexta) larvae differ in sensitivity to polycationic peptides. AB - We have recently characterized specific binding sites for human interferon-gamma on particulates prepared from the protocerebrum and hemolymph of tobacco hornworm larvae, Manduca sexta ?(Parker, M.S., Ourth, D.D., 1999. Comp. Biochem. Physiol. B 122, 155-163). The sensitivity to sulfated polysaccharides indicated an involvement of oligobasic epitopes of hIFN-gamma in the binding. In the present study, we found that polycationic peptides inhibited the binding of [125I]hIFN gamma to particulates from either the hemolymph or the protocerebrum of Manduca sexta larvae. With amino acid homopolymers, the rank order of potency was poly-L lysine > poly-L-arginine >> poly-L-ornithine, while the acidic side chain polymer poly-L-aspartate was not inhibitory. However, the potency of all polycationic peptides was at least three-fold greater at the hemolymph particulates. Also, acidic polysaccharides such as heparin were much more efficacious in the inhibition of hIFN-gamma binding to hemolymph relative to protocerebral particulates. The peptide polycations inhibited the binding of [125I](Leu31,Pro34)human peptide YY, a ligand selective for the Y1 subtype of the neuropeptide Y receptor, to rabbit kidney or to parietal cortex particulates with the expected rank order of poly-L-arginine > poly-L-lysine >> poly-L-ornithine, and with little cross-tissue difference in affinity. The selectivity observed with M. sexta particulates indicates a preferential involvement of oligobasic lysine-rich C-terminal sequences of IFN-gamma, while large insect tissue-related affinity differences point to involvement of diverse oligoacidic sequences in binding to protocerebrum and hemolymph sites. This study provides evidence for the presence of molecules in lepidopteran larvae that are similar in structure to vertebrate co-receptors of IFN-gamma, and adds to the characterization of these binding sites. PMID- 10818268 TI - The muscle fatty acid binding protein of spadefoot toad (Scaphiopus couchii). AB - Fatty acid binding protein was purified from skeletal muscle of the spadefoot toad (Scaphiopus couchii), an estivating species. While estivating, this animal relies on the fatty acid oxidation for energy. Hence we were interested in the behaviour of fatty acid binding protein under conditions of elevated urea (up to 200 mM) and potassium chloride such as exist during estivation. Also we examined whether there were interactions between glycolytic intermediates and the binding ability of the protein. The amount of bound fatty acid (a fluorescence assay using cis-parinarate) was not affected (P < 0.05) by glucose, fructose 6 phosphate or phosphoenolpyruvate at physiological concentrations. By contrast, glucose 6-phosphate increased the amount of bound cis-parinarate but the apparent dissociation constant was not different from the control. Fructose 1,6 bisphosphate but not fructose 2,6-phosphate decreased cis-parinarate binding by 40%, commensurate with doubling the apparent dissociation constant (1.15-2.62 microM). Urea, guanidinium and trimethylamine N-oxide at 200 mM increased cis parinarate binding 60% over controls. Urea (1 M) and KCl (200 mM) did not affect cis-parinarate binding compared to controls. The interaction of this fatty acid transporter with fructose 1,6-bisphosphate is discussed in terms of reciprocal interaction with phosphofructokinase since fatty acid is also an inhibitor of phosphofructokinase. PMID- 10818269 TI - Storage metabolism in the Pacific oyster (Crassostrea gigas) in relation to summer mortalities and reproductive cycle (west coast of France). AB - We describe seasonal changes in the biochemical composition of digestive gland, adductor muscle and gonad and surrounding mantle area in Crassostrea gigas from the Western Atlantic coast of France. Seasonality in histology of storage tissues and glycogen storage capacity in isolated vesicular cells were also studied. Proteins, the main muscle components did not contribute to the gametogenetic effort. Glycogen and lipids were stored in the digestive gland, gonad and surrounding mantle area during the wintering period and the gonad and surrounding mantle area represented the main storage compartment supplying the reproductive effort. Gametogenesis in spring and summer was associated with an increase in lipid and protein contents and took place at the expense of glycogen reserves. Histological study of storage tissue in the gonad led us to define four seasonal stages of storage tissue development. In vitro, glycogen storage capacity in isolated vesicular cells was high from November to March and markedly reduced during gametogenesis, decreasing below detectable levels after spawning. This physiological state should be taken into account with relation to summer mortalities occurring in commercial growing areas. PMID- 10818270 TI - A comparison of plasma vitamin C and E levels in two Antarctic and two temperate water fish species. AB - Antarctic fish have a high polyunsaturated lipid content and their muscle cells have a high mitochondria density suggesting that Antarctic fish are under greater oxidative stress than temperate water fish. To test this hypothesis, the plasma concentrations of the antioxidant vitamins E and C were measured in two Antarctic fish species, Pagothenia borchgrevinki and Trematomus bernacchii, and compared with the plasma concentrations of these vitamins in two New Zealand temperate water fish species, blue cod (Parapercis colias) and banded wrasse (Notolabrus fucicola). Neither vitamin is known to be synthesised in fish and so must be obtained from the diet. The plasma from both Antarctic fish species had vitamin E concentrations five to six times higher than those found in the two temperate water fish species. However, significantly higher levels of vitamin C were only found in the plasma of T. bernacchii, a benthic Antarctic fish. The average level of vitamin C in the plasma of the cryopelagic P. borchgrevinki was approximately one-third that of T. bernacchii. The T. bernacchii plasma yielded a high range of vitamin C values, possibly reflecting differences in nutritional status among the animals captured. No beta-carotene was found in any of the fish plasma samples studied. The data suggest that even though Antarctic fish live at -1.5 degrees C they may be exposed to greater metabolic stress from free radical mediated oxidation than temperate water species. PMID- 10818271 TI - Cloning of the Atlantic salmon (Salmo salar) estrogen receptor-alpha gene. AB - A cDNA clone encoding most of an Atlantic salmon (Salmo salar) estrogen receptor (ER) was obtained from a liver cDNA library and the remainder of the coding sequence from the gene was isolated from a genomic library. Sequence comparisons showed that the cloned gene represents ER-alpha. Expression of the ER-alpha gene in male and female salmon parr was analysed by RT-PCR. Highest expression was found in brain and liver, with lower levels of ER-alpha mRNA present in all other tissues tested. There was little difference in expression of ER-alpha between male and female. PMID- 10818272 TI - Hepatic glucose phosphorylating activities in perch (Perca fluviatilis) after different dietary treatments. AB - Increased activity of hepatic glucose phosphorylation was observed in perch after feeding previously fasted fish. When a pellet diet containing 14% carbohydrate was given, most of the increased activity had a low affinity towards glucose (S0.5 = 19.5 mM) and resembled the mammalian glucokinase (Hexokinase IV or D) and the glucokinase-like activity previously observed in salmon liver. In addition, increased activity of a hexokinase with high affinity towards glucose (Km = 0.50 mM) was observed with the pellet diet. An increase in the activity of this hexokinase alone was observed when the fish were fed with filet of cod containing less than 0.2% carbohydrate. Perch with a very high hepatic glucokinase-like activity after eating the pellet diet had high activities of pyruvate kinase and glucose-6-phosphate dehydrogenase, indicating a high capacity of glycolysis and carbohydrate utilization. Simultaneously, the activity of glycogen phosphorylase was strongly reduced while the activity of fructose-1,6-bisphosphatase was not significantly changed. These observations were made with perch captured in the spawning season and brought to the laboratory. Assays of glucose phosphorylation in livers of perch eating the natural diet (insects) in the lake showed no glucokinase-like activity. PMID- 10818273 TI - Astaxanthin and its metabolites idoxanthin and crustaxanthin in flesh, skin, and gonads of sexually immature and maturing Arctic charr (Salvelinus alpinus (L.)). AB - Carotenoid compositions of the flesh, skin, and ovaries were determined in sexually maturing and immature Arctic charr (Salvelinus alpinus) fed diets supplemented with astaxanthin (optical isomer ratio (3S,3'S):(3R,3'S; meso):(3R,3'R); 1:2:1). Astaxanthin comprised 64-79% of the flesh carotenoids, and the 3',4'-cis and 3',4'-trans glycolic isomers of idoxanthin, present in a 1:1 ratio, represented 20-35%. The flesh of the sexually maturing charr contained relatively more idoxanthin than that of sexually immature fish (20 vs 35% of total carotenoids), possibly being indicative of a higher metabolic turnover of astaxanthin in the latter. The relative proportions of flesh carotenoids were unaffected by sex. The relative carotenoid composition of ovaries was similar in sexually maturing and immature females. The 3',4'-cis and 3',4'-trans glycolic isomers of idoxanthin (ratio 0.7:1) were the major carotenoids (56% of total), followed by crustaxanthin (20%), and astaxanthin comprised less than 5% of ovarian carotenoids. Three glycolic isomers of crustaxanthin were detected (3,4,3',4'-di-cis-:3,4-cis-3',4'-trans-:3,4,3',4'-di-trans-glycolic isomer ratio 2.6:3.1:1) in the ovaries. Sex and maturity status had no apparent effect on the relative composition of skin carotenoids. The skin carotenoids consisted mainly of diesters (82-87% of total carotenoids) and monoesters (7-13% of total carotenoids). Saponification revealed that astaxanthin comprised 85% and idoxanthin 10% of total carotenoids, and minor amounts of tunaxanthin-, lutein-, and zeaxanthin-like metabolites were also present. Maturity status seems to be more important than sex in determining the relative carotenoid composition of the tissues of Arctic charr, with astaxanthin and its metabolites being selectively accumulated in different tissues. PMID- 10818274 TI - Proinsulin cDNAs from the leopard frog, Rana pipiens: evolution of proinsulin processing. AB - We have isolated a proinsulin cDNA from the Amphibian Rana pipiens. The predicted R. pipiens insulin A- and B-chain amino acid sequences differ from that deduced from the closely related Rana catesbeiana at one residue (Asp for Pro at B2). The R. pipiens and Xenopus laevis proinsulin precursor sequences are of identical length, with the amino acid sequences of the mature A- and B-chains being well conserved. The proinsulin C-peptide amino acid sequence is less well conserved between R. pipiens and X. laevis and also differs in length. The R. pipiens C peptide is shorter than the homologous X. laevis sequence due to a two amino acid residue truncation. The truncation of the R. pipiens C-peptide compensates for a two amino acid residue extension observed at the N-terminal of the A-chains of insulins from Ranid frogs. A change in the site of proinsulin processing can explain both the C-peptide and A-chain length differences. The evolution of the new proinsulin processing site required two amino acid substitutions. PMID- 10818275 TI - Permeability of the infective juveniles of Steinernema carpocapsae to glycerol during osmotic dehydration and its effect on biochemical adaptation and energy metabolism. AB - Permeability of the sheath and cuticle of the infective juveniles (IJs) of Steinernema carpocapsae to glycerol and its effect on biochemical adaptation of the IJs to osmotic dehydration were examined by incubating both sheathed and exsheathed IJs in glycerol-d5 solution then monitoring the changes in levels of deuterium labelled and non-labelled glycerol and trehalose. Energy metabolism of the IJs during osmotic dehydration and subsequent rehydration and the effect of the permeated glycerol on this process were investigated by examining and comparing the changes in mean dry weight and key biochemical composition of the IJs dehydrated in glycerol and sodium chloride solutions. The results show: (1) similarly to evaporative dehydration, osmotic dehydration induces IJs to synthesise the protectants glycerol and trehalose; (2) glycerol permeates the sheath and the cuticle into the body of IJs during dehydration in glycerol solution. Part of the permeated glycerol plays a role as protectant like that synthesised by IJs from their energy reserve materials while part is incorporated into trehalose; (3) the sheath reduces the rate of permeation of glycerol and therefore affects the equilibrium glycerol and trehalose levels of the IJs and also the time needed to reach the equilibrium levels; (4) the reduction in mean dry weight and lipids of the IJs during dehydration in glycerol solution is substantially less than those dehydrated in sodium chloride solution. Both the total protectant level and the ratio of glycerol to trehalose of the IJs dehydrated in glycerol solution are higher than those dehydrated in sodium chloride solution; (5) glycogen reserves of the IJs play a role as a buffer reservoir of the protectants during both dehydration and rehydration but the principal sources of the protectants during dehydration are more likely to be lipids and proteins rather than glycogen. PMID- 10818276 TI - Occurrence of ommochrome-containing pigment granules in the central nervous system of the silkworm, Bombyx mori. AB - Dark-red pigment granules were found in the brain and ganglion of the normal strain of the silkworm, Bombyx mori, by light microscopy. No other pigmentation was seen in the brain or ganglia. Electron microscopy showed that the granules were electron-dense. The granules were similar to the ommochrome-containing pigment granules that are present in the epidermal cells of the quail mutant, as previously reported. The pigment in the larval central nervous system (CNS) of the normal silkworm was identical to the ommin standard with respect to the absorption spectrum, the infrared spectrum, and the Rf value in thin-layer chromatography (TLC). After acid hydrolysis of the pigment, 3-hydroxykynurenine was detected by TLC. The pigment granules in the CNS contained mainly ommin. An ommochrome-binding protein was also detected in the CNS by in vitro binding studies and Western blotting. The ommochrome granules may have an important function in the CNS of the silkworm. PMID- 10818277 TI - Activity and localisation of the lysosomal marker enzymes acid phosphatase, N acetyl-beta-D-glucosaminidase, and beta-galactosidase in the earthworms Eisenia fetida and E. veneta. AB - The present study describes the activity and localisation of three putative lysosomal marker enzymes, acid phosphatase (AP), N-acetyl-beta-D-glucosaminidase (beta-NAG), and beta-galactosidase (beta-Gal), in whole individuals and in distinct parts of the earthworms, Eisenia veneta and Eisenia fetida. Activities of AP and beta-NAG were high in the two species with most of the activity located to the anterior and mid-parts of the worms. The activity of beta-Gal was low in all body regions. We found interspecies difference in the AP activity as E. veneta had significantly higher activity of AP than E. fetida in posterior and mid-parts, as well as in whole individuals. Of the three enzymes tested, AP was the only enzyme located to lysosomes, yielding high latency all over the worms with especially high latency in the coelomic fluids and posterior regions. The lysosomal APs in E. veneta and E. fetida may be utilised as a new biomarker for xenobiotic-induced lysosomal membrane damage in earthworms. PMID- 10818278 TI - Blunted temperature and cortisol responses to ipsapirone in major depression: lack of enhancement by electroconvulsive therapy. AB - Depression has been shown in some studies to be associated with a reduction in hypothalamic 5-HT(1A) receptor function, as indicated by reduced hormone and/or hypothermic responses to 5-HT(1A) agonists such as ipsapirone. The hypothermic response to ipsapirone was reduced in depressed patients treated with amitriptyline. Hormone and hypothermic responses to 5-HT(1A) agonists were reduced in normal subjects administered specific serotonin reuptake inhibitors. Effects of electroconvulsive therapy (ECT) on 5-HT(1A) receptor-mediated responses in humans have not been reported. In the present work, ten depressed patients and 15 control subjects were challenged with placebo and with 0.3 mg/kg ipsapirone, administered 48 h apart in a randomised double blind design. Hypothermic, growth hormone (GH) and cortisol responses were measured. Seven of the depressed patients were treated with a course of ECT, and placebo and ipsapirone challenges were repeated 24 and 72 h after the last treatment. The cortisol response to ipsapirone was significantly reduced in the depressed patients compared with controls. The hypothermic response to ipsapirone was totally abolished in the depressed patients. When tested after a course of ECT, the seven depressed patients again showed reduced or blunted responses. We conclude that hypothalamic 5-HT(1A) receptor function is reduced in depression. In contrast to the effects of electroconvulsive shock (ECS) on post-synaptic 5 HT(1A) receptor function in animals, which have chiefly been measured in the hippocampus using electrophysiological techniques, ECT in humans does not induce an increase in sensitivity of post-synaptic 5-HT(1A) receptors in the hypothalamus. PMID- 10818279 TI - Modulation of neuroendocrinological function by psychosomatic training: acute effect of ChunDoSunBup Qi-training on growth hormone, insulin-like growth factor (IGF)-I, and insulin-like growth factor binding protein (IGFBP)-3 in men. AB - The neuroendocrine system is regarded as the major link between the psychological state and the immune system in man. The present study was undertaken to examine the acute effect of ChunDoSunBup Qi-training, a traditional psychosomatic training, on the plasma level of growth hormone (GH), insulin-like growth factor (IGF)-I and insulin like growth factor binding protein (IGFBP)-3 in young men. To characterize the plasma level of hormones and the pattern of IGFBP-3 at pre- (-10 min), mid-time (40 min), and immediately after (post-time, +10 min) Qi-training, radioimmunoassay (RIA) and western ligand blot (WLB) analysis were used. The plasma level of GH at the mid-time was significantly increased (P<0.05) compared to the level at pre-time of Qi-training. The plasma level of IGF-I was also increased at mid-time (P<0.05) and immediately after Qi-training. There was a significant correlation between the levels of GH and IGF-I immediately after Qi training (r=0.69, P<0.01). In this study, the change of IGFBP-3 among the several IGFBPs was determined. There was a relative shift in IGFBP-3 for the 43-kDa fraction during mid-time of Qi-training (P<0.05). These results suggest that Qi training influences the growth hormone, IGF-I and IGFBP-3 status via brain-target organ axis in men. PMID- 10818280 TI - Leptin and IGF-I levels in unconditioned male volunteers after short-term exercise. AB - We have previously shown that serum gonadotropins, particularly LH, decline after acute exercise in male volunteers. The mechanism for this decline is unknown. Plasma leptin and IGF-I concentrations were measured in seven male volunteers after acute exercise to exhaustion using the Bruce protocol. Leptin concentrations declined following exercise reaching nadir values 30-120 min after exercise. As anticipated, plasma IGF-I concentrations showed a transient rise immediately after exercise falling thereafter to nadir levels 60-90 min after exercise before returning towards baseline levels. In view of the previously described decline in gonadotropin release after acute exercise, the decline in plasma leptin levels, perhaps related to the rise in IGF-I, may play a role in exercise-induced inhibition of gonadotropin release presumably by inhibition of GnRH secretion. PMID- 10818281 TI - Reduced adrenocortical responsiveness to adrenocorticotropic hormone (ACTH) in socially subordinate female marmoset monkeys. AB - Socially subordinate female common marmoset monkeys undergo pronounced, chronic reductions in basal plasma cortisol levels, which appear to result both from socially induced suppression of reproductive hormones and from direct effects of social subordination. In this study, we tested the hypothesis that this cortisol suppression is mediated by reduced adrenocortical responsiveness to adrenocorticotropic hormone (ACTH). Dominant, subordinate, and ovariectomized females were given dexamethasone (5 mg/kg, IM), followed the next morning by human ACTH(1-39) (10 microg/kg, IV) or sterile saline (0.5 ml/kg, IV); blood samples were collected at -20 through 150 min from ACTH or saline treatment and assayed for cortisol. ACTH, but not saline, caused a marked elevation of plasma cortisol levels. Prior to ACTH treatment, dominant females tended to have higher dexamethasone-suppressed cortisol levels than subordinate and ovariectomized females. After ACTH treatment, dominant females had significantly higher cortisol concentrations, as well as higher peak and net integrated cortisol responses to ACTH, than did subordinate and ovariectomized animals; the latter two groups showed comparable cortisol responses to ACTH. These results suggest that dampened adrenocortical responsiveness to ACTH contributes to chronic reductions in cortisol levels in subordinate female marmosets and may be mediated by suppression of reproductive hormones. PMID- 10818282 TI - Neuroendocrine correlates of temperamental traits in humans. AB - Studies investigating temperament traits in humans and their biological correlates have found high levels of novelty seeking (NS) linked with dopaminergic system changes, and particularly a deficit of dopamine transporter. Harm avoidance and reward dependence, on the other hand, appeared to be associated, respectively with serotonin and noradrenaline changes. In the present study, we have investigated the dopaminergic (DA), serotonergic (5-HT), and noradrenergic (NE) functions in healthy volunteers by challenging the monoamine systems with the DA agonist bromocriptine, the 5-HT agonist D-fenfluramine, and the NE agonist clonidine, respectively. Parallel to this investigation, we examined the temperament traits of our subjects by measuring NS, harm avoidance (HA) and reward dependence (RD) using the 'Three-dimensional Personality Questionnaire' (TPQ). The aims of the study were to see whether or not the monoamine functions were correlated with temperament traits. Bromocriptine challenge induced a significant GH increase and a significant suppression of PRL. D-fenfluramine test significantly increased PRL and cortisol plasma levels and Clonidine test induced a significant rise in GH values. NS scores showed a significant direct correlation with brom-stimulated GH values (r=0.426, P<0.05) and a significant inverse correlation with brom-inhibited PRL values (r=-0.498, P<0.01). HA scores correlated significantly with D-fen-stimulated PRL and CORT AUCs, (PRL: r=0.424, P<0.05; CORT: r=0. 595, P<0.005). RD scores correlated positively with clon-stimulated GH values (r=0.55; F=8.6; P<0.01) and negatively with brom-inhibited-PRL AUCs (r=-0.439, P<0.05). Our data support Cloninger theory concerning the biological correlates of temperamental traits, and evidence the link between the neuroendocrine responses to dynamic challenges and stable temperament features. PMID- 10818283 TI - Language lateralization and handedness in women prenatally exposed to diethylstilbestrol (DES). AB - Hand preferences and language lateralization were assessed in women exposed prenatally to the synthetic estrogen, diethylstilbestrol (DES), and in their unexposed sisters. The DES-exposed women showed an increased degree of hand preference (regardless of direction) and were more likely to be left handed for writing. However, the groups did not differ significantly on a dichotic listening measure of language lateralization. Perhaps as a result of the alterations in hand preferences, the typical relationship between hand preferences and language lateralization was disrupted in the DES-exposed group. Also, within the DES exposed group, exposure early in gestation correlated with left handedness whereas exposure late in gestation correlated with reduced left ear (right hemisphere) scores on the verbal dichotic task. Results are discussed in terms of theoretical perspectives predicting hormonal influences on sexual differentiation of hemispheric asymmetry and in terms of separate critical periods for hormonal effects on individual sexually differentiated characteristics. PMID- 10818284 TI - Increased activity of the hypothalamus-pituitary-adrenal system after treatment with the mineralocorticoid receptor antagonist spironolactone. AB - The hippocampal mineralocorticoid receptor (MR) is critical for the regulation of the basal activity of the hypothalamus-pituitary-adrenocortical (HPA) system. It has been hypothesized that reduced capacity of the hippocampal MR is involved in the HPA-system dysregulation found in depression and aging. We applied the combined dexamethasone suppression/corticotropin releasing hormone stimulation (DEX/CRH) test to six healthy young females both before and after 12 days of treatment with the MR antagonist spironolactone to assess HPA regulation. Treatment with spironolactone caused a significant increase in post-dexamethasone cortisol concentrations (75.1+/-56.7 vs. 36.6+/-24.6 nmol/l, p<0.05). Furthermore, we observed a significant rise in peak cortisol concentration after additional human CRH (hCRH) application (223. 6+/-139.1 vs. 126.7+/-73.3 nmol/l, p<0.02). There was no change in ACTH plasma concentrations. We thus conclude that (1) the MR antagonist spironolactone affects HPA system regulation as reflected in the DEX/CRH test and (2) these findings are in accordance with the assumption that MR dysfunction may underlie HPA-system dysfunction in depression and/or aging. PMID- 10818285 TI - GABA levels within the medial preoptic area: effects of chronic administration of sodium valproic acid. AB - Sodium valproic acid (VPA) is a widely prescribed anticonvulsant medication that has been shown to interfere with pubertal maturation of the reproductive system, and induce endocrine abnormalities in adults, within a subset of the clinical population. While VPA's mechanism of action is still poorly understood, it may exert its anti-reproductive effects by enhancing GABAergic inhibition of the GnRH neuronal population within the medial preoptic area (mPOA). The purpose of this study was to determine if chronic administration of VPA alters GABA levels within the mPOA region. In Experiment 1, the mPOA, caudate, and arcuate nucleus regions were harvested from VPA-treated and control mice. Analysis of whole tissue content of GABA revealed that levels were lower in the caudate and arcuate nucleus regions of VPA-treated animals, whereas there were no group differences for the mPOA region. Collapsing across drug group, there was also a trend for males having overall higher levels of GABA as compared to females. In Experiments 2 and 3, mice were implanted with microdialysis probes within the mPOA region and sampled for extracellular GABA levels. Females (Exp. 3) were sampled either on diestrous, proestrous, or estrous. Results from males (Exp. 2) revealed that VPA enhanced extracellular GABA levels in the mPOA region compared with controls. However, GABA levels for both groups remained stable across the sampling period. Conversely, in Exp. 3, females showed cyclical release of GABA across the sampling period. For control females, GABA levels increased during the afternoon on all cycle days, but the rise on proestrus was smaller than on other cycle days. VPA-treated animals showed an overall reduction in GABA levels compared with controls. Furthermore, while GABA increased over sampling time on estrus and diestrus days of the cycle, there was not a significant rise in GABA on proestrus. These data indicate: (1) regional specificity in VPA effects upon GABA levels, (2) a sex difference in the effects of VPA on GABA levels within the mPOA, and (3) GABA levels increase on the afternoon of all days of the estrous cycle with VPA attenuating the rise seen on the afternoon of proestrus. These results provide evidence that VPA effects upon the reproductive axis may involve changes in GABA release, and that males and females show different patterns of neurochemical response to the drug. PMID- 10818297 TI - Between and within breed variation in lamb survival and the risk factors associated with major causes of mortality in indigenous Horro and Menz sheep in Ethiopia. AB - Data collected on 3256 lambs born to Horro and Menz breed ewes single-sire mated to 71 rams at the International Livestock Research Institute (ILRI) Debre Berhan station between September 1992 and June 1996 were analysed for rates of survival and growth from birth to weaning. A significantly lower proportion of Menz lambs died before 1 year of age (28%) than the Horro lambs (59%). Least squares means for pre- and post-weaning mortality were 8.8 and 19.3%, respectively in Menz, and 25.3 and 34.2% for Horro sheep. Major causes of death were similar in Horro versus Menz lambs and were pneumonia (53 vs. 54%, respectively), digestive problems (14 vs. 12%), endoparasite infections (9 vs. 13%), starvation mismothering-exposure (SME) complex (10 vs. 7%) and septicemia (3 vs. 2%). Relationships among causes of mortality with breed, birth weight (BWT), season of birth, parity, litter size and lamb health category (number of times a lamb was sick between birth and 1 year of age) were determined. The impact of these factors on mortality varied with lamb age. Lambs that were born with <2kg BWT had a greater risk of dying from any cause except pneumonia. But, even though Horro lambs were heavier than Menz at birth (2.4 vs. 2.1kg, respectively), twice as many died before 1 year of age. The cause of mortality was further influenced by season of birth, lamb sex and health category. In addition, sires were a significant source of variation for progeny survival at 6, 9 and 12 months of age, but not at the younger ages. The best and worst Horro ram sired progeny groups that had mortality rates up to 1 year of age of 22 vs. 80%, respectively. The same estimates in Menz rams were 11 and 48%, respectively. Reduced mortality rate would significantly increase lamb output. However, isolated efforts to solve this problem are likely to have limited impact. Instead, an integrated approach to minimise the impact of underlying factors is advocated. Farm (animal) management routines that could be introduced in the short or longer term are discussed. PMID- 10818298 TI - Comparative electrocardiographic studies, and differing effects of pentazocine on ECG, heart and respiratory rates in young sheep and goats. AB - The effects of pentazocine on electrocardiogram (ECG) heart and respiratory rates were studied in young sheep and goats as a prelude to its clinical use as an opioid analgesic in the two species. The ECG was monitored by standard bipolar leads that provided discernible electrocardiographic features only in Lead I. Electrocardiograms of lambs differ significantly from those of kids with respect to QRS complex, P-R interval and T wave. Higher basal heart rate in lambs is apparently due to shorter P-R interval and shorter interval between successive cardiac cycles (p<0.05). Pentazocine, 3mgkg(-1) i.m., appears safer for lambs than kids as it significantly decreased P-R interval, Q-T interval, cardiac cycle duration, interval between successive cardiac cycles, respiration rate, and increased heart rate in kids but not in lambs (p<0.05). PMID- 10818299 TI - Microscopic and PCR findings in sheep after experimental infection with Ehrlichia phagocytophila. AB - Methods available for the diagnosis of Ehrlichia phagocytophila infection are limited and of low sensitivity. Since polymerase chain reaction (PCR) is a highly sensitive and specific method for the detection of infectious agents in animals, it was considered useful to develop a PCR method specific for this agent. An experimental protocol in which blood from 10 different flocks was inoculated into 10 naive 1 month-old lambs was carried out. Progress of the infection was monitored during 3 weeks by daily rectal temperature and blood samples. Eight lambs showed hyperthermia, presence of E. phagocytophila in blood, and lower growth rate while the other two never showed the agent in their blood or any clinical signs. This experimental infection, essentially designed to provide infected samples, also showed that flocks from areas previously thought to be free of E. phagocytophila were actually infected. PCR on blood samples allowed detection of the infection earlier than in case of microscopic examination. This was related to an increased sensitivity (87-91%). PCR yielded positive results on samples with as few as 1.08 infected neutrophils/ul (about 0.1%). PMID- 10818300 TI - The pattern of ciliation and the development of the epithelial lining of the respiratory tract in the neonatal kid: a scanning electron microscopic study. AB - The pattern of ciliation and the development of the epithelial lining of the respiratory tract was investigated in 20 Cashmere kids, aged between 6h to 21 days. Samples were taken from the alar and basal folds, nasal conchae, nasal septum, nasopharynx, epiglottis, vocal fold, infraglottic cavity, trachea, extrapulmonary bronchus, and parenchyma to include respiratory bronchi and alveoli. The samples were routinely prepared and evaluated with scanning electron microscopy and light microscopy. The results showed that cilia were more densely packed and more extensively distributed within the rostral region of the nasal cavity of the kids. Large patches of non-ciliated microvillous cells usually seen in adult goats were not a feature of the kid. It was also noted that the epithelium covering the nasal septum was heavily ciliated in newborn to 3-day-old kids. From that age onwards, the epithelium became deciliated to attain a 'moth eaten' appearance. Bronchioles in kids were poorly developed, with respiratory bronchioles being rarely encountered in the first week of life. A cell type characterized by a wrinkled, large apical surface presenting short, apical microvilli, was frequently observed in the larynx and trachea of the kid. Lung parenchyma frequently presented evidence of alveoli formation, whereas alveolar pores were less numerous. PMID- 10818301 TI - Effects of dietary energy density on feed intake, body weight gain and carcass chemical composition of Omani growing lambs. AB - Forty male Omani lambs were used in a feeding trial to study the effects of feeding diets containing various levels of metabolizable energy (ME) on growth and carcass composition. Ten lambs were selected randomly and slaughtered at the start of the trial to provide information on initial carcass composition. Thirty lambs were divided randomly into three groups and fed three diets varying in ME concentration (low, medium and high) from weaning (at average 76 days) until slaughter at the mean weight of 30kg. Digestibility of dry matter (DM) was 66.9, 68.7 and 73.9% for low, medium and high energy diets, respectively. Apparent gross energy digestibility was 66.8, 67.2 and 73.3% corresponding to dietary concentrations of 12.2, 12.6 and 13.9MJ of DE/kg for low, medium and high energy diets, respectively. Daily DM intake ranged between 3.12 and 3.73 % of body weight (BW) which was equivalent to 76.5-97.5g/kg(0.75) or 0.738-1.142MJ ME/kg(0.75). Daily BW gain increased (P<0.001) with increasing ME density with a maximum of 154g/day observed in lambs on high energy diet during the last 4 weeks of the experiment. Feed conversion ratio (FCR), i.e., kg feed/kg BW, improved with increasing ME density (P<0.001). Sheep fed high energy diet had heavier BW (P<0.01), empty BW weight (P<0.001), carcass weight (P<0.01) higher dressing percentage (P<0.05) but lower gut content (P<0.001) than lambs fed medium and low energy diets. Sheep slaughtered at the end had lower water, protein but higher carcass and non-carcass chemical fat than sheep slaughtered at the start of the experiment. This study indicated that meat production from sheep in Oman will be improved in form of higher BW gains and better carcass composition by increasing energy levels in the diet. PMID- 10818302 TI - Pharmacokinetics and penetration into tissue fluid of ceftizoxime in normal and hyperthermic sheep. AB - The pharmacokinetics of ceftizoxime was studied in six sheep before and after inducing hyperthermia using Escherichia coli endotoxin. Sheep implanted subcutaneously with cages of non-reactive material for collecting tissue cage fluid (TCF) were used to conduct two trials. In Trial 1 animals with normal basal temperature (normal sheep (NS)) were given intravenous (i.v.) and intramuscular (i.m.) monodoses of ceftizoxime (20mg/kg BW) at 1 week interval. One and 5 weeks later (Trial 2) each sheep were injected 1ug/kg BW of endotoxin to produce hyperthermia (hyperthermic sheep (HS)) previously to i.v. administration (HSi.v.) and i.m. (HSi.m.) of ceftizoxime (20mg/kg BW), respectively. Serum and TCF samples were collected over 6h post-administration. Ceftizoxime concentrations in serum and TCF were determined by a microbiological assay. The concentrations in serum and TCF of ceftizoxime were analyzed through compartmental and non compartmental models.Rectal temperature were significantly incremented in all animals during Trial 2. The half-time and constant of elimination in serum of ceftizoxime in NSi.v. (t(1/2)=1.1+/-0.4h; lambda(z)=0.7+/-0.2h(-1)) were statistically different those observed in HSi.v. (t(1/2)=1.4+/-0.4h; lambda(z)=0.5+/-0.2h(-1)). The constants of distribution in NSi.v. and HSIV were 5.1+/-4.6 and 4.1+/-3.4h(-1), respectively. The time to reach the maximum concentrations in TCF was latter (p<0.05) in NS (t(max)=2.3+/-0.7h) than in HS (t(max)=1.3+/-0.6 h). After i.m. administration in NS the absorption half-life (0.12+/-0.19h) was latter (p<0.05) than in HS (0.06+/-0.007h) with greater areas under the curve (AUC in NS=65.4+/-20.8 and AUC in HS=34.7+/-7.5 (ug/ml) h). The maximum value of concentration in serum (C(max)) and AUC in TCF were greater (p<0.05) in NS (C(max)=46.1+/-10.6ug/ml and AUC=84.4+/-17.4 (ug/ml) h) as compared to same HS (C(max)=27.0+/-12.9ug/ml and AUC=47.9+/-3.9 (ug/ml) h). The concentrations of ceftizoxime in TCF after i.v. and i.m. in NS and HS were elevated during a 6h period after administration. The bioavailability of ceftizoxime in NS (101.6+/-59.9%) and HS (87.4+/-63.3%) was suitable for its use by the i.m. route. PMID- 10818303 TI - Effect of naloxone on the plasma levels of LH, FSH, prolactin and testosterone in Beetal bucks. AB - Ten adult male Beetal goats were used for the study to elucidate the modulation of gonadotrophin, prolactin and testosterone secretion by endogenous opioid peptides. An indwelling catheter was placed in the jugular vein of each buck 20h before the onset of the experiment. Bucks were divided randomly into two groups: Group I (n=5) received naloxone at a dose rate of 1mg/kg body weight (BW) and Group II (n=5) received naloxone at a dose rate of 2mg/kg BW intravenous. Blood samplings were done from 2h before treatment until 2h after treatment at 15min intervals. Blood samples were quantified for plasma LH, FSH and prolactin concentration using a heterologous double antibody radioimmunoassay (RIA) and testosterone concentration was quantified by coat-a-count RIA kit. The mean plasma LH levels during pretreatment phase were 0.41+/-0.03ng/ml in Group I and 0.44+/-0.02ng/ml in Group II which significantly (p<0.05) increased to 0.91+/ 0.05ng/ml in Group I and 1.53+/-0.07ng/ml in Group II. The mean plasma FSH levels did not show a difference in pre- and post-treatment animals in both groups. A significant (p<0.05) increase in plasma testosterone concentration was observed in both groups after naloxone treatment, whereas, a decrease (p<0.05) was observed in plasma prolactin levels after naloxone treatment. Thus, it can be concluded that endogenous opioids do play an important role in modulating plasma LH, prolactin and testosterone concentrations in male goats. PMID- 10818304 TI - Embryo recovery and transfer for the production of transgenic goats from Korean native strain, Capra hircus aegagrus. AB - During the breeding season in Korea (September 1997 to April 1998), a goat embryo recovery and transfer program using a Korean native strain (Capra hircus aegagrus) was performed for the production of transgenic goats. Donors were synchronized with norgestomet implants and superovulated by a combined treatment with FSH and hCG. The treatment regime induced a consistent and efficient superovulation rate of 90% from donors with an ovulation rate of 12.1+/-0.5. 50.5% of the recovered oocytes/embryos were fertilized and most of them were at the 1-cell stage. After DNA microinjection, a total of 188 embryos were immediately transferred to naturally cycling or hormonally synchronized recipients with two or three embryos per animal. There was a tendency for the pregnancy rate of naturally cycling recipients to be higher (38.9%) than that of hormonally synchronized recipients (25.7%). When the recipients were classified into a two or three embryo-received group, the embryo viability was markedly decreased from 58.3% in two embryo-received group to 35.3% in three embryo received group, without an increase in the kidding rate. This resulted from a high occurrence of abortions or stillbirths from the multiple-pregnant recipients which had received three embryos. This indicated that the transfer of two embryos per recipient is highly recommendable for an optimal embryo survival in Korean goats. Altogether, 188 embryos were transferred to 71 recipients and two transgenic Korean goats were produced from 25 offspring. PMID- 10818305 TI - Evaluation of cellular and biochemical parameters of blood and peritoneal fluid following enterectomy in the goat. AB - To evaluate the effects of enterectomy on cellular and biochemical parameters of blood and peritoneal fluid, an experiment was conducted using 10 Iranian crossbred male goats. Ten milliliter of blood and 1-1.5ml of peritoneal fluid were sampled from all animals prior to operation for the estimation of control values. Enterectomy was performed under local anesthesia. Blood and peritoneal fluid samples were collected at 24, 48, 72 and 96h after enterectomy. The results revealed that after enterectomy, the number of WBCs, neutrophils, monocytes and band neutrophils in the blood significantly increased (p<0.05). However, the percent of lymphocytes decreased significantly (p<0.05). The concentrations of glucose and blood urea nitrogen (BUN) significantly increased (p<0.05). Furthermore, following the operation, the number of WBCs and the percent of neutrophils and band neutrophils in the peritoneal fluid significantly increased (p<0.05). In contrast, the percent of lymphocytes and monocytes in the peritoneal fluid decreased significantly (p<0.05). The concentrations of protein and the activities of amylase and alkaline phosphatase (ALP) in the peritoneal fluid increased significantly (p<0.05). However, the concentration of glucose decreased significantly (p<0.05). This study showed that enterectomy can have profound effects on blood and peritoneal fluid parameters. PMID- 10818306 TI - Monitoring of estrous cycle of ewes by ram-seeking behavior. AB - For 6 years adult, non-synchronized Hampshire ewes (n=2,5,8,8,9 and 12) and rams (n=6,10,12,11,11 and 9) were maintained separately and released to contiguous pastures having in common a woven-wire fence about 80m long and 1.1m high. Natural estrous cycles were monitored daily by observations of ram-seeking behavior as defined as a ewe being persistent in staying at the common fence with attention focused on the ram to the virtual exclusion of grazing. Day 0 of the estrous cycle is the first day that ram-seeking is observed. One hundred and thirty four fence stays and 115 estrous cycles have been observed. For cycles falling within the normal range, the mean cycle duration (+/-SE) was 16.53+/-0.03 days. During the first four study years, a copulation test with an intact ram, carried out on a spot-check basis, indicated that all tested ewes showing ram seeking, except one, were in estrus. Ram-seeking never occurred outside of the estrous period of the cycle. During the fifth year, daily copulation tests with an intact ram and daily observations for ram-seeking has indicated that estrus generally lasts about two days while ram-seeking lasts for one of these days, so that the two behaviors while overlapping are not coextensive. During the sixth year, daily copulation tests with a vasectomized ram showed again a partial coincidence of the two behaviors, but also that occasionally they appeared to have the same duration. It is concluded that the utilization of ram-seeking provides an alternative, practical means to monitor the estrous cycle. PMID- 10818307 TI - Production responses of two genetic different types of Merino sheep subjected to different nutritional levels. AB - The primary selection objective in the South African Mutton Merino is meat production, while wool plays a secondary role. The Merino, on the other hand is selected for wool production. Respectively 20 and 23 of each genetic different type of ewes were used in a 15-week indoor study to quantify the influence of nutrient intake on certain production traits when fed at different levels of their respective requirements during late pregnancy (last 6 weeks) and lactation (first 6 weeks). Results were used to provide production parameters for the two types of ewes and their lambs at different levels of nutrition. Lambing percentage (per ewe lambed) of SA Mutton Merino ewes was 171% compared to 135% for Merino ewes. Live weight gains for meat-wool and wool-type ewes during late pregnancy amounted to 15.9 and 21.9%, with corresponding live weight losses of 8.3 and -1.1% during the first 6 weeks of lactation. Greasy wool production during the last 9 weeks of pregnancy and the first 6 weeks of lactation amounted to 1.57kgewe(-1) (meat-wool type ewes) and 2.84kgewe(-1) (wool-type ewes). Milk production 17 days post partum was 2.33kg/day and 1.84kg/day for the two types of sheep. Lambs of the meat-wool type ewes had a mean birth weight of 4.1kg and an average daily gain (ADG) of 270g/day to 42 days of age. Lambs of the wool-type ewes had a mean birth weight of 4.9kg and a 42-day ADG of 192g/day. There were clear differences in production traits of the two types of Merino sheep, which were in accordance with their genetic differences. The study gave practical production norms for meat-wool and wool-type Merino ewes and their lambs when subjected to different nutritional levels. PMID- 10818308 TI - Carcass characteristics of llamas (Lama glama) reared in Central Chile. AB - Body and carcass composition were studied on 10 male and 10 female naturally reared llamas (Lama glama). Half the animals were young (9-12 months) and the other half adult (>3 year). The average live weights for young and adult males were 104.4 and 100.6kg, and for females 67.6 and 104.6kg, respectively. Average carcass weights for the four groups were 58.9, 56.2, 36.8 and 56.7kg, respectively. Carcass composition for males and females was similar, but males had slightly higher dressing percentages than females (56.1 and 55.8 vs. 54.1 and 54.2 for young and adult males and females, respectively). Carcass length and fat depth at the loin and proportions of cuts in the carcass were similar for both the sexes, except for leg and tail, which were proportionately heavier in young females compared to the other groups. The composition of meat on fresh basis was: moisture 70.2%, protein 20.5%, ether extract 8.23% and ash 3.4%. Age and sex seemed to have no effects on the body and carcass characteristics studied nor on the chemical composition of meat. PMID- 10818309 TI - Effect of parity on milk yield, composition, somatic cell count, renneting parameters and bacteria counts of Comisana ewes. AB - Twenty-four Comisana ewes, with no history of mastitis, were included in this study, with eight ewes each in parities 1, 2 and 3. Groups were separately penned on straw litter and ewes were individually checked for yield, composition, renneting properties and bacteriological characteristics of milk from January, when separated from their lambs (50+/-3 days after lambing), to May. Samples with more than 3.5x10(5) somatic cells/ml were cultured for mastitis related pathogens. Milk yield was not significantly affected by parity. The P3 ewes had significantly higher milk protein, casein and fat contents compared to the P1 and P2 ewes. The P3 ewes also had improved renneting ability of milk as compared to the P1 ewes. Quality of milk decreased with lower lactations. The milk of P1 ewes had significantly greater amounts of mesophilic bacteria than the P2 and P3 ewes, as well as higher concentrations of psychrotrophs and total coliforms in their milk with respect to the P3 ewes. Somatic cell counts in milk and the prevalence of subclinical mastitis were not changed by parity, although mastitis infection set in progressively earlier as the number of lactations decreased. These results suggest that ewes in first or second lactation have a less favourable milk secretion status in relation to mastitis than ewes with a higher number of lactations. Milk yield and quality of younger ewes may be improved by offering feed rations that take into account this reduced capacity to mobilise body reserves. Also, most scrupulous control of sanitation of housing, equipment and personnel is necessary. PMID- 10818310 TI - Sexual activity and body and testis growth in prepubertal ram lambs of Friesland, Chios, Karagouniki and Serres dairy sheep in Greece. AB - Sexual activity, body size and testis growth in the prepubertal period (120 210days of age) were studied in five male lambs of each of four dairy sheep breeds. Sexual responses developed earlier in Friesland than in Chios and Karagouniki and latest in Serres ram lambs. Friesland lambs expressed interest in females at 127days of age and 34kg live weight 18-36days younger and 3.4-6.3kg lighter than the three Greek breeds. Puberty (first mounting with ejaculation occurred) of ram lambs appeared to be more closely associated with body weight than with chronological age. Age at puberty averaged 179days in Friesland, 8, 10 and 30days younger (P<0.05) than in Karagouniki, Chios and Serres, respectively. On the other hand, weight at puberty averaged 50kg for Friesland and Chios compared to about 44kg for the two other breeds (P<0.05). In the four breeds studied there was a 41-52days interval between the time when lambs first expressed interest in females and the time puberty occurred, the longest was that of Friesland lambs. There was a significant breed difference in scrotal circumference. At puberty the scrotal circumference was 33.9cm in Friesland lambs, 4-6cm greater than in the three Greek breeds. Friesland ram lambs had a greater scrotal circumference throughout the entire period studied. PMID- 10818311 TI - Comparison of mineral and cholesterol composition of different commercial goat milk products manufactured in USA. AB - Concentrations of 12 major and trace minerals and cholesterol in commercial goat fluid milk, evaporated, powdered, yogurt, and cheese products manufactured in the US were evaluated for compositional differences. Minerals were determined by an Inductively Coupled Argon Plasma Emission Spectroscopy (ICAP), while cholesterol was analyzed using colorimetric and gas chromatographic (GC) methods. Mean total solids content (%) of fluid milk, evaporated milk, powdered milk, yogurt, plain soft and Monterey Jack cheeses were: 11.3, 20.9, 94.1, 11.5, 32.5, and 57.7, respectively. Mean calcium and phosphorus contents (ppm, dry basis) of the corresponding products were: 103, 125; 440, 393; 7715, 7471; 161, 144; 691, 1105; 3492, 3067, respectively. The respective iron and zinc contents (ppm) of the corresponding products were: 0.062, 0.349; 1.518, 1.635; 3.33, 30.21, 0.117, 0.338; 7.16, 3.64; 8.86, 3.81. The levels of potassium (K) in cheeses were lowest among all the products including fluid goat milk, suggesting that a significant amount of K was lost during cheese manufacturing processes. Levels of all trace minerals were higher in yogurt and cheeses than in fluid milk. The levels of trace minerals in cheeses were greater than those in yogurt products. Iron and aluminum contents of certain goat milk products were considerably higher than normal, possibly due to contamination of minerals from manufacturing utensils and product metal cans. Sulphur contents of fermented products were significantly greater than those of fluid milk, which may be accountable for the microbial synthesis of sulfur containing proteins during the manufacturing processes of the products. Cholesterol contents (mg/100g, wet basis) of fluid, evaporated, powdered goat milk and Monterey Jack cheese determined by GC method were: 11.0, 24.9, 119.5 and 91.7, respectively. Cholesterol contents of the goat milk products analyzed by colorimetric method were substantially greater than those by GC method. PMID- 10818312 TI - Seasonal variation in semen characteristics of Chios and Friesian rams in Greece. AB - Five Chios and five Friesian rams were used to study the effect of season on semen production. Semen samples were collected with an artificial vagina. In spite of the large variation of most of the semen characteristics among rams of the two breeds, seasonal variation in semen quantity (volume, concentration and total number of spermatozoa per ejaculate) and quality (percentage of motile spermatozoa, sperm progressive motility and percentage of abnormal spermatozoa) were significant (P<0.05). The best semen was produced mainly during autumn and the worst during spring (semen volume: 1.47ml in Chios and 1.48ml in Friesian rams versus 1.24ml in Chios and 1.07ml in Friesian rams; percentage of abnormal sperm: 5.66% in Chios and 5.42% in Friesian rams versus 8.22% in Chios and 7.75% in Friesian rams; total sperm/ejaculate (x10(9)): 7.05 in Chios and 6.74 in Friesian rams versus 4.09 in Chios and 4.05 in Friesian rams). The magnitude of these seasonal effects was not sufficient to prevent rams from being used for breeding throughout the year. However, the existence of differences among rams within each breed in semen quality and quantity makes it necessary to perform a semen evaluation on individual basis in order to select the best males before they are used for breeding. PMID- 10818313 TI - Comparative performance of Awassi lambs and Black goat kids on different crude protein levels in Jordan. AB - An experiment was conducted to investigate the effect of different crude protein (CP) levels on the growth performance of Awassi lambs and Black goat kids. Forty lambs and 40 kids were randomly selected and allocated to separate pens of equal size for 75 days. Each group was fed one of four rations containing 12, 14, 16, or 18% CP. Final weights and gains for lambs were higher (P<0.001) than those for kids for all rations fed. No differences were obtained between lambs fed with 16 and 18% CP ration. However, both groups had higher (P<0.001) body weight and gain than lambs in other groups. Kids had higher (P<0.001) body weights and gain when fed with 16% ration. There were no significant differences among kids of other groups. Black goat kids have good potential to support red meat production in Jordan with less efficiency than Awassi lambs. PMID- 10818314 TI - Preliminary survey for antibodies against caprine arthritis-encephalitis virus (CAEV) using recombinant GAG proteins: studies among small ruminant populations in north-eastern Nigeria. AB - A total of 1000 serum samples were obtained from small ruminants in Maiduguri, Borno state, Nigeria and tested for the presence of antibodies against caprine arthritis-encephalitis virus (CAEV) using enzyme-linked immunosorbent assay (ELISA) based on p17 and p28 recombinant GAG proteins. The distribution of the sera tested was as follows: 900 serum samples collected at slaughter from 700 goats and 200 sheep in the municipal abattoir as well as 100 sera obtained from 50 each of goats and sheep in four different flocks under the semi-intensive system of animal husbandry. All the animals sampled were aged >/=2 years and had no previous contact with imported stocks. It was observed that none of the sera had antibody against CAEV. The need to impose strict quarantine as well as the practice of testing and slaughtering of positive animals imported from CAEV endemic areas into Nigeria for breeding are suggested to prevent the introduction of the disease into the country. PMID- 10818315 TI - Sheep-goat hybrid born under natural conditions. AB - The paper reports a 5 year old male sheep-goat interspecific hybrid born under natural conditions. The hybrid was castrated at 10 months of age. Karyotype analysis confirmed that the animal was a male and a hybrid with 57 chromosomes, an intermediate number between sheep (2n=54) and goat (2n=60). Morphological characteristics of the animal were compared with those of castrated male Tswana goats of the same age. Data on castrated male Tswana sheep of the same age are not available. The hybrid at 5 years of age weighed 93kg compared to 53.73+/ 13.83kg, the average weight of castrated Tswana male goats of the same age. The paper also raises a question of whether the animal could be of commercial interest in meat production in Botswana. PMID- 10818316 TI - The effect of water availability on feed intake and digestion in sheep. AB - Six castrated male Karagouniko sheep, offered three levels of water, were used in a 3x3 Latin Square design to study the effects of mild water restriction on roughage intake and digestion. The three treatments were offered water ad libitum throughout the day (A), made available to the animals for 1h daily (B) or 65% of the water consumed ad libitum (C). Sheep in Treatment A consumed more water (181.9g/kg BW(0.75)) than the two water restriction regimes, which did not differ significantly (B: 128.8, C: 117.5g/kg BW(0.75)) between them. Roughage dry matter intake and digestibility of nutrients did not differ significantly between treatments. Non-significant differences (p>0.05) were also observed in the rate of passage of undigested residues between treatments (A: 48.7, B: 56.9 and C: 54.5 %/h). It was concluded that the water restrictions usually applied during the dry season did not have any significant effect on the nutrition of a local breed of sheep in Greece. PMID- 10818317 TI - Effects of dietary sulfur level on amino acid concentrations in ruminal bacteria of goats. AB - 12 Angora (18+/-0.6kg BW) and 20 Alpine (24+/-1.0kg BW) goat wethers consumed diets (14.3% CP and 1.67-1.80Mcal/kg ME, DM basis) with 0.11, 0.20, 0.28 or 0.38% S (supplemental S:CaSO(4); N:S ratio is 21, 12, 8 and 6, respectively) for 10 weeks to determine effects of dietary S on amino acid concentrations in ruminal fluid bacteria. The concentration of cysteine in bacterial DM changed quadratically (P<0.08) as dietary S increased (3.28, 3.77, 3.80 and 3.65% for 0.11, 0.20, 0.28 and 0.38% S, respectively). However, dietary S did not alter methionine concentration in bacterial DM or total amino acids, and for the few amino acids whose concentrations were affected, magnitudes of change were relatively small. In conclusion, with diets moderate to low in ME, levels of S greater than 0.20% and N:S ratios less than 12:1 had very little effect on amino acid concentrations in ruminal fluid bacteria of growing goats, which supports the contention that the primary potential influence of inorganic dietary S on absorbed S-containing amino acids is through the quantity of microbial protein synthesized in the rumen. PMID- 10818318 TI - Effect of two doses of progesterone on estrus response and fertility in acycling crossbred Bharat Merino ewes in a semi-arid tropical environment. AB - The efficacy of two doses of progesterone (P4), i.e. 350 and 300mg was evaluated in acycling crossbred ewes (50) during the autumn breeding season. Ewes were treated with intravaginal progesterone sponges with either 350mg or 300mg for 12 days. At standing estrus, ewes were hand mated (three cycles). Progesterone (350mg) gave a better (P<0.05) estrus response (75%), compared to 300mg dose (42%). Ewes treated with 350mg P4 also showed better (P<0.05) synchronization response (93%) than those treated with 300mg (56%) at 72h after sponge removal. Lower doses of progesterone (300mg) significantly delayed (P<0.01) the onset of estrus. However, dose had no significant influence(P>0.05) on estrus length, conception rate and the proportion of ewes lambing at term. This study indicates that dose of progesterone may have an effect on estrus exhibition and response time without altering the conception rate and lambing in acycling crossbred Bharat Merino ewes during the major breeding season in a semi-arid tropical environment. PMID- 10818319 TI - Repeatability and relationship with field mating performance of a serving capacity pen test in rams. AB - Two studies were conducted to analyse the repeatability of a serving capacity (SC) pen test and to evaluate the correlation of the results of such a test with field performance. The Laborde SC test, which measures SC by exposing an individual ram for 40min to two restrained non-oestrous ewes in a pen was used. In the first study, SC was measured on four and six occasions during the autumn of 1994 and 1995, respectively. The results indicated that this test is reliable in measuring SC of rams, since SC was repeatable within, and between, breeding season of consecutive years (p<0.05). In the second study conducted on two commercial farms, two groups of ewes synchronised with intravaginal sponges containing medroxyprogesterone acetate, were served by rams of high or low SC. On both the farms, the group of ewes served by rams of high SC had higher pregnancy rates than those served by low SC rams (p<0.05). The Laborde test proved to be a reliable test for the evaluation of SC in rams and that high SC rams had better field performance. PMID- 10818320 TI - Uniport soft-tip, open-ended catheters versus multiport firm-tipped close-ended catheters for epidural labor analgesia: a quality assurance study. AB - STUDY OBJECTIVE: To compare a multiport, firm-tipped, close-ended, epidural catheter (Portex catheter) with a uniport, open-ended, soft-tipped, wire reinforced catheter (Arrow catheter) in obstetric patients. STUDY DESIGN: Prospective data collection for intradepartmental quality assurance. SETTING: Obstetric unit in a tertiary care maternity hospital. PATIENTS: 2612 patients requesting labor analgesia. INTERVENTIONS: The Arrow catheter was used in 1,352 women and the Portex catheter in 1,260 women. MEASUREMENTS AND MAIN RESULTS: The incidence of unsatisfactory block were 3.3% and 4.4% with the Arrow and Portex catheters, respectively (p = 0.2). The catheter perforated the dura matter in 0.4% of cases with both catheters. The incidence of epidural venipuncture was 1.1% with the Arrow catheter and 5.7% with the Portex catheter (p = 0.0001). Paresthesias occurred in 6% of cases with the Arrow catheter and 11. 2 % of cases with the Portex catheter (p = 0.0001). Epidural catheter reinsertion was required in fewer patients in the Arrow group than in the Portex group (4.8% vs. 7.1%; p = 0.01). CONCLUSIONS: In obstetric patients, the softer uniport Arrow catheter produces paresthesias and venipunctures less frequently than the firm multiport Portex catheter. PMID- 10818321 TI - Determination of eicosanoid and cytokine production in salvaged blood, stored red blood cell concentrates, and whole blood. AB - STUDY OBJECTIVE: To determine the production of the eicosanoids prostaglandin 2 (PGE2) and thromboxane 2 (TxB2) and the cytokines interleukin 1 beta (IL-1-beta) and interleukin 6 (IL-6) in whole blood (WB), unfiltered red blood cell (RBC), and filtered RBC concentrates, and salvaged blood. DESIGN: Prospective study. SETTING: University hospital of Erlangen. PATIENTS: 32 healthy volunteers and 14 ASA physical status I, II, and III radical prostatectomy patients (mean age 65 yrs). INTERVENTIONS: Sixteen WB units and 16 RBC units (divided into 16 filtered and unfiltered units each) were taken from 32 volunteers. Fourteen salvaged RBC units were obtained from the 14 radical prostatectomy patients. Sixteen WB units were stored for 35 days. From the 16 WB donations, RBC concentrates (PAGGS-M) were prepared. The RBC concentrates were halved, one half had its leukocytes removed at day 0; both halves were stored for 49 days. Salvaged blood (n = 14) was stored up to 2 hours during surgery and then retransfused. MEASUREMENTS AND MAIN RESULTS: Immediately at the start of the study, in all blood units (WB, RBC filtered, and RBC unfiltered units) at days 0 and 21, and at the end of the storage period (WB: 35 days, RBC concentrates: 49 days) and in the salvaged RBC units, the following parameters were measured: PGE2, TxB2, IL-1-beta, IL-6, hematocrit, platelet number, leukocytes, blood volume, and hemoglobin. During storage, different levels of PGE2, TxB2, IL-1-beta, IL-6 for WB, filtered RBC concentrates, and unfiltered RBCs were found. The higher levels of PGE2, TxB2, IL 1-beta, and IL-6 were found in the WB and RBC salvaged units than the filtered RBCs or unfiltered RBC units. There was no statistically significant difference between WB and salvaged RBCs. Higher levels of leukocytes and platelets were found in WB units and salvaged RBCs as compared to filtered or unfiltered RBCs. CONCLUSIONS: The eicosanoid and cytokine levels in the salvaged, filtered RBC, unfiltered RBC, and WB units stayed within physiological limits, suggesting that these levels do not contribute to the risk of nonhemolytic, immunomodulated transfusion reactions, even in massive transfusions. PMID- 10818322 TI - Coasting after overpressure induction with sevoflurane. AB - STUDY OBJECTIVE: To evaluate the clinical feasibility of using a coasting technique to temporarily maintain anesthesia after overpressure induction with sevoflurane. STUDY DESIGN: Prospective clinical study. SETTING: Large teaching hospital. PATIENTS: 12 ASA physical status I, II, and III patients receiving general anesthesia for a variety of peripheral procedures. INTERVENTIONS: After overpressure induction of anesthesia with sevoflurane (8%) in an O(2)/N(2)O mixture, the fresh gas flow (FGF) was lowered to 0.5 L/min and the vaporizer was turned off (coasting). MEASUREMENTS AND MAIN RESULTS: After priming a circle system with sevoflurane (8% sevoflurane vaporizer setting in 6 L/min O(2)/N(2)O [33%/66%] for 30 s), patients took several vital capacity breaths from the mixture until loss of consciousness. After 3.4 +/- 0.7 min, depth of anesthesia was considered adequate for laryngeal mask airway (LMA) insertion, and FGF was reduced to 0.5 L/min (33% O(2), 66% N(2)O) and the sevoflurane vaporizer was turned off. The end-expired sevoflurane concentration (Et(sevo)) decreased from 5.8 +/- 1.3% just before insertion of the LMA to 0.97 +/- 0.22% at 20 minutes. CONCLUSIONS: After overpressure induction with sevoflurane, coasting during minimal flow anesthesia (FGF 0.5 L/min) is a simple technique that can maintain anesthesia for short procedures (less than 15 to 20 min), or can be used as a bridge or an adjunct to other low-flow techniques. PMID- 10818323 TI - Awareness during laryngoscopy and intubation: quantitating incidence following induction of balanced anesthesia with etomidate and cisatracurium as detected with the isolated forearm technique. AB - STUDY OBJECTIVE: To measure the incidence of awareness during induction of anesthesia with etomidate and fentanyl, and to model its frequency as a function of dose of etomidate. DESIGN: Prospective cohort study. SETTING: Anesthesia department of a university hospital. PATIENTS: 30 ASA physical status I, II, and III patients undergoing elective general surgery. INTERVENTIONS: Patients were assigned to one of three groups of etomidate (0.2 mg/kg, 0.3 mg/kg, 0.4 mg/kg) and received fentanyl (2 microg/kg) and 2 x ED(95) of cisatracurium (0.1 mg/kg). Neuromuscular block was monitored with a peripheral nerve stimulator. Intubation was performed after maximum T(1)-depression. To identify awareness, the isolated forearm technique (IFT) was used. The IFT was performed by prompting the patient every 20 seconds. Only a verified response was considered a positive IFT response. Anesthesia was maintained with isoflurane in oxygen/air and fentanyl. MEASUREMENTS AND MAIN RESULTS: Maximum neuromuscular block occurred after 352 +/- 96 seconds and intubation was performed 424 +/- 86 seconds after loss of consciousness (LOC). Awareness was dose dependent: 80% of patients receiving 0.2 mg/kg etomidate, 70% of patients receiving 0.3 mg/kg etomidate, and 20% of patients receiving 0.4 mg/kg etomidate had a positive IFT response. Awareness occurred in one patient 3 minutes after LOC, in 65% during laryngoscopy, and in 30% within the following 120 seconds. One patient had explicit recall without finding awareness unpleasant. Hemodynamic parameters did not differ between patients with a positive or a negative IFT response. CONCLUSIONS: The incidence of awareness during bolus induction can be modeled as dose dependent. However, when combining a short-acting induction drug and a delayed-onset neuromuscular blocker, the continuous infusion of the hypnotic drug may prevent awareness during induction. PMID- 10818324 TI - Gender is an essential determinant of blood transfusion in patients undergoing coronary artery bypass graft procedure. AB - STUDY OBJECTIVE: To determine factors that account for gender difference in the need for blood transfusion in coronary artery bypass graft (CABG) patients. DESIGN: Retrospective study of consecutive patients. SETTING: Anesthesiology department of a teaching hospital. PATIENTS: 253 CABG patients (163 males and 90 females). INTERVENTIONS: Packed red blood cells (PRBCs), platelets, and fresh frozen plasma (FFP) were transfused depending on the need of each patient. MEASUREMENTS AND MAIN RESULTS: For each patient, we recorded the gender, age, weight, height, body surface area (BSA), and duration of surgery. Hematocrit (Hct) levels prior to surgery, end of surgery, and at discharge from the hospital were recorded. PRBC administration and use of FFP and platelets were noted. Differences between the data for female and male patients were evaluated using Student's t-test, Chi-square test, and regression analysis. Approximately 60% female and only 20% male patients received PRBCs intraoperatively, whereas 78% females and only 43% males received PRBCs during their entire hospital stay. On average, females received 1.20 units of PRBCs intraoperatively and 2.38 units during the entire hospital stay, while the males received 0.31 units and 1.36 units for similar periods. Gender differences in PRBC transfusion persisted even when females and males were compared within the same subgroups for age, weight, duration of surgery, and preoperative Hct. PRBC units given intraoperatively had a significant correlation with age and preoperative Hct in females, but they had a significant correlation with age, preoperative Hct, and duration of surgery in males. PRBCs given during the entire hospital stay, however, had significant correlation with age, preoperative Hct, and duration of surgery in both females and males. Multiple logistic regression analysis showed that the probability of a patient receiving or not receiving PRBC transfusion is significantly influenced by age, preoperative PRBC mass, duration of surgery, and gender. CONCLUSION: Gender is an independent essential determinant of blood transfusion in CABG patients, and it may interact with age, weight, preoperative Hct, duration of surgery, and other factors in determining the probability of transfusion. PMID- 10818325 TI - Cardiovascular homeostasis during inhalational general anesthesia: a clinical comparison between sevoflurane and isoflurane. On behalf of the Italian Research Group on sevoflurane. AB - STUDY OBJECTIVE: To obtain more information on cardiovascular homeostasis and patient discharge from the recovery area after general anesthesia with either sevoflurane or isoflurane as the main anesthetic. DESIGN: Prospective, randomized, multicenter study. SETTING: Inpatient anesthesia at 13 University Departments of Anesthesia. PATIENTS: 247 ASA physical status I, II, and III patients, aged 18 to 85 years, receiving general anesthesia for elective urological, orthopedic, ENT, vascular, and low abdominal surgery. INTERVENTIONS: General anesthesia was maintained using a 60% nitrous oxide in oxygen mixture with either isoflurane (n = 125) or sevoflurane (n = 122) adjusted according to hemodynamic variables. MEASUREMENTS AND MAIN RESULTS: Occurrence of hypotension [systolic arterial blood pressure (SBP) decrease >30% from baseline], hypertension (SBP increase >30% from baseline), bradycardia [heart rate (HR) <50 bpm], or tachycardia (HR>100 bpm) provoked stepwise changes in the inspired concentration of the study drug. If this action proved to be ineffective after an adequate stabilization period, a specific treatment was given, and the need for pharmacological treatment was recorded as a hemodynamic side effect by an independent observer. No differences in duration of anesthesia were observed between sevoflurane (126 +/- 76 min) and isoflurane patients (139 +/- 60 min). Mean duration from anesthetic discontinuation to fulfillment of discharge criteria was shorter after sevoflurane (21 min; 25(th) to 75(th) percentiles: 27 to 13 min) than isoflurane (27 min; 25(th) to 75(th) percentiles: 17 to 35 min) (p < 0.0005). Hemodynamic side effects requiring therapy occurred in 18 sevoflurane patients (14.6%) and 26 isoflurane patients (20.8%) (p = NS). The risk for hemodynamic side effects increased with age (>50 vs. < or OFFyrs: odds ratio 2.5; 95% CI 1.2 to 5.4; p = 0.015) and ASA physical status (III vs. I and II: odds ratio 2.2; 95% CI 0.9 to 5.7; p = 0.048). When only patients over 50 years of age were considered (72 in the sevoflurane group and 79 in the isoflurane group), the incidence of hemodynamic side effects was higher with isoflurane (29.1%) than with sevoflurane (15.2%) (odds ratio 2.3; 95% CI 1.0 to 5.2; p = 0.04). CONCLUSIONS: Sevoflurane provided equally safe and effective control of cardiovascular homeostasis as isoflurane, with a more rapid discharge from the recovery area. Interestingly, patients over 50 years of age showed a lower risk for hemodynamic side effects when receiving sevoflurane than isoflurane. PMID- 10818326 TI - Effects of the phosphodiesterase-III inhibitor enoximone on skeletal muscle specimens from malignant hyperthermia susceptible patients. AB - STUDY OBJECTIVES: To study the in vitro effects of the phosphodiesterase-III inhibitor enoximone on skeletal muscle specimens from malignant hyperthermia susceptible (MHS) and normal (MHN) patients. DESIGN: Prospective study. SETTING: Malignant hyperthermia (MH) laboratory at a university hospital. PATIENTS: 47 patients with clinical suspicion for MH undergoing in vitro contracture test (IVCT) for diagnosis of MH susceptibility. INTERVENTIONS: Biopsies of M. quadriceps femoris were performed in adult patients with a 3-in-1 nerve block and in children with trigger-free general anesthesia. MEASUREMENTS AND MAIN RESULT: Patients were first classified as MHS or MHN by the IVCT according to the protocol of the European MH Group (EMHG). Patients with equivocal results (MHE) or with neuromuscular diseases were excluded from the study. Enoximone was added to the organ bath to surplus vital muscle specimens in single bolus concentrations of 0.4, 0.6, 0. 8, or 1.6 mmol/L. The in vitro effects of enoximone on muscle contractures and twitch were measured. Seventeen patients were classified as MHS and 30 as MHN by the EMHG criteria. Enoximone induced contractures in skeletal muscles in a dose-dependent manner. Contractures of MHS compared to MHN muscle specimens were significantly larger at all concentrations used in this study. No overlap in maximum contractures was seen between MHS and MHN muscles at a bath concentration of 0.6 mmol/L enoximone only. CONCLUSIONS: Diagnosis of MH by an IVCT test with a single bolus administration of enoximone seems to be possible using a concentration of 0.6 mmol/L. The findings of this study may indicate an involvement of the phosphodiesterase-III and cAMP system in pathogenesis of MH. Further in vivo investigation should determine the trigger potency of enoximone in MH susceptible individuals. PMID- 10818327 TI - A multicenter comparison of isoflurane and propofol as adjuncts to remifentanil based anesthesia. AB - STUDY OBJECTIVE: To compare recovery, hemodynamics, and side effects of remifentanil-based anesthesia with hypnotic concentrations of isoflurane or propofol. DESIGN: Multicenter, prospective, randomized, two-group study. SETTING: 15 university and 5 municipal hospitals. PATIENTS: 249 ASA physical status I, II, and III adult patients scheduled for elective gynecological laparoscopy, varicose vein, or arthroscopic surgery of at least 30 minutes' duration. INTERVENTIONS: Anesthesia was induced in the same manner in both groups: remifentanil-bolus (1 microg/kg), start of remifentanil-infusion (0. 5 microg/kg/min), followed by propofol as needed for induction. Five minutes after intubation, remifentanil was reduced to 0.25 microg/kg/min, and it was combined with either a propofol infusion (0.1 mg/kg/min) or with isoflurane (0.6 vol% end-tidal) in O(2)/air. Adverse hemodynamic responses of heart rate and systolic blood pressure were recorded and treated according to a predefined protocol. With termination of surgery, anesthetic delivery was discontinued simultaneously without tapering, and recovery times were recorded. MEASUREMENTS AND MAIN RESULTS: No significant differences were observed between the remifentanil-isoflurane or remifentanil propofol treatment regimens. Recovery times (means +/- SD) were similar for spontaneous ventilation (5.8 +/- 3.2 min vs. 6. 3 +/- 3.7 min), extubation (7.6 +/- 3.5 vs. 8.5 +/- 4.2 min), eye opening (6.8 +/- 3.2 vs. 7.5 +/- 3.8 min), and arrival to the postanesthesia care unit (16.5 +/- 7.0 vs.18.0 +/- 7.2 min). There were no significant differences in adverse hemodynamic responses, postoperative shivering, nausea, or vomiting between the groups. CONCLUSIONS: Emergence after remifentanil-based anesthesia with 0.6 vol% of isoflurane is at least as rapid as with 0.1 mg/kg/min propofol. Both isoflurane and propofol are suitable adjuncts to remifentanil, and the applied dosages are clinically equivalent with respect to emergence and recovery. Therefore, both combinations should be appropriate, particularly in settings in which rapid recovery from anesthesia is desirable, such as fast tracking and/or ambulatory surgery. PMID- 10818328 TI - Myocardial ischemia correlates with reduced fibrinolytic activity following peripheral vascular surgery. AB - STUDY OBJECTIVES: To evaluate the relationship between perioperative ischemia and serial concentrations of D-dimer, which is a sensitive and specific marker of fibrinolytic activity. Myocardial ischemia and infarction are well-recognized complications of peripheral vascular surgery. We hypothesized that patients at increased risk of perioperative myocardial ischemia might be identified preoperatively by abnormal hemostatic indices. DESIGN: Prospective clinical outcomes study. SETTING: A 1,124-bed tertiary care medical center. PATIENTS: 42 ASA physical status II, III, and IV patients undergoing peripheral vascular surgery. INTERVENTIONS: Serial D-dimer concentrations were measured preoperatively, and at 24 and 72 hours postoperatively. Continuous 12-lead ST segment monitoring (Mortara Instrument, Inc., Milwaukee, WI) was performed with the acquisition of a 12-lead ECG every 20 seconds for 72 hours. MEASUREMENTS AND MAIN RESULTS: D-dimer measurements were performed in duplicate using the Dimer Gold assay (American Diagnostica, Greenwich CT). Ischemic episodes, as defined by continuous 12-lead ST-segment monitoring, occurred in 49% of patients. There were no demographic differences between ischemic and nonischemic groups. Although baseline D-dimer concentrations were not statistically significantly different between groups, patients experiencing perioperative myocardial ischemia generated significantly less D-dimer during the perioperative period (p = 0. 014). CONCLUSIONS: PATIENTS with an impaired fibrinolytic response, as defined by reduced generation of D-dimer, experienced an increased incidence of perioperative myocardial ischemia. PMID- 10818329 TI - A randomized study of left versus right internal jugular vein cannulation in adults. AB - STUDY OBJECTIVE: To compare the success rate and incidence of complications of right internal jugular vein (RIJV) versus left internal jugular vein (LIJV) cannulation using external landmarks or surface ultrasound guidance. DESIGN: Prospective randomized study. SETTING: Operating room of a university-affiliated hospital. PATIENTS: 120 adult patients scheduled for elective abdominal, vascular, or cardiothoracic procedures with general anesthesia and mechanical ventilation in whom central venous cannulation was clinically indicated. INTERVENTIONS: Patients were randomized to four groups for RIJV cannulation using the landmark approach (Group 1) or surface ultrasound (Group 2) versus LIJV cannulation with the landmark approach (Group 3) or ultrasound (Group 4). MEASUREMENTS AND MAIN RESULTS: The data collected included time from first puncture to guidewire insertion, number of attempts, and associated complications. If conversion to the ultrasound technique was required, the number of crossover patients and reasons for failure were recorded. Cannulation of the LIJV was more time consuming; it required more attempts; and it was associated with a greater number of complications when compared to the right side (p < 0.05). CONCLUSIONS: Left IJV cannulation is more time consuming than RIJV cannulation and is associated with a higher incidence of complications. The use of ultrasound improves success rate and decreases the number of complications during IJV cannulation. PMID- 10818330 TI - Risk attitudes of anesthesiologists and surgeons in clinical decision making with expected years of life. AB - STUDY OBJECTIVE: To understand anesthesiologists' and surgeons' risk attitudes about expected years of life. DESIGN: Prospective study. SETTING: Central operating rooms (ORs) in a university hospital. RESPONDENTS: 122 anesthesiologists and surgeons. MEASUREMENTS AND MAIN RESULTS: A 7-page survey questionnaire regarding the length of survival and the choice of treatments was sent to 122 physicians. Certainty equivalents and Arrow-Pratt coefficients of absolute risk aversion (APC) were calculated from the survey. Of 122 physicians, 93 responded (38 anesthesiologists and 55 surgeons; response rate 76. 2%). There was no significant difference in risk attitudes between the anesthesiologists and the surgeons. Their age was a statistically significant predictor of the risk attitudes: the older they were, the greater their APCs were (i.e., the more risk averse they were). There was no significant difference between gender in risk attitudes, nor was there any significant difference between the risk attitudes of becoming patients and those of working in the OR. CONCLUSIONS: Physicians' risk attitudes cannot be predicted by their specialties or gender. However, the older they are, the more risk-averse they are. PMID- 10818331 TI - Equipment for the difficult airway in obstetric units in Germany. AB - STUDY OBJECTIVE: To examine the availability of specialized equipment for the difficult airway management in obstetric units of German departments of anesthesiology. DESIGN AND SETTING: An anonymous questionnaire survey was mailed to the directors of 993 German departments of anesthesiology. Completed replies were grouped by number of deliveries performed each year. MAIN RESULTS: 55.5% of the hospitals responded. Data of 449 answers were evaluated for this investigation. A difficult airway cart was available in 99.3% of the departments. More detailed investigation revealed that different shaped laryngoscope blades (74.9% of the departments), laryngeal masks (91.0%), a fiberoptic bronchoscope (85.9%), and transtracheal puncture devices (59.9%) were available in the majority of the units. However, only a minority of the departments had these devices directly available in their obstetric operating rooms (OR; laryngeal masks 36.2%, fiberoptic bronchoscope 23.9%, transtracheal puncture set 22.0%). Larger units with more than 1,000 deliveries per year provided their equipment more often directly in the obstetric OR or the facility housing the obstetric unit than did smaller units with less than 1,000 deliveries per year (p< 0.001). CONCLUSIONS: The survey of German departments of anesthesia revealed that specialized equipment for the difficult airway management often is not directly available in the obstetric OR. Anesthesiologists must familiarize themselves as to which difficult airway equipment is available in their unit and where it is stored. PMID- 10818332 TI - Living liver donor surgery: report of initial anesthesia experience. AB - The charts and anesthetic records of 12 patients who donated the left lateral segment of their liver to a related infant or child to treat liver failure were retrospectively reviewed. Blood loss, need for transfusion, fluids administered, surgical length, and perioperative complications were investigated. The records also were examined to determine the hemodynamic stability of patients undergoing donor hepatectomy to assess their need for invasive monitoring. There were no episodes of hypotension or hemodynamic instability. The average operating time was 9.6 +/- 1.1 hours. The blood loss was 562 +/- 244 mL (range 300 to 1100 mL). Four patients received their own cell saver blood (200 mL, 220 mL, 300 mL, 475 mL), and one patient received 1 U (350 mL) of predonated autologous blood. The average hemoglobin decreased significantly (p = 0.001) from a preoperative value of 14.1 +/- 1.2 to 12.3 +/- 1.8 g/dL in the recovery room. All patients were extubated in the operating room or recovery room. Patients were discharged home in 6.9 +/- 1.3 days (range 5 to 9 days). Living-related liver resection can be performed with noninvasive monitoring and without the need for heterologous blood products. PMID- 10818333 TI - Sudden bronchospasm on intubation: latex anaphylaxis? AB - I present a case of a patient with a history of cerebral palsy and asthma, living in a group home, who developed acute onset bronchospasm immediately after intubation. The patient developed hypotension 5 minutes after intubation. The bronchospasm lasted 20 minutes, and the case was complicated further by continued hypotension and a pneumothorax. A diagnosis of latex-mediated anaphylaxis was made in the intensive care unit after immunoglobin E (IgE), serum tryptase, and latex-specific IgE antibody were shown to be markedly elevated. This case report demonstrates that immediate onset of bronchospasm on intubation of an asthmatic patient is not always an asthma attack, and that other causes of bronchospasm should be considered in the differential diagnosis. Patients with a history of atopy, including those with a history of asthma, have an increased risk of developing latex sensitivity. It is important to remember that more than one etiology may be responsible for this kind of bronchospasm, and that it may be difficult to differentiate between multiple etiologies of bronchospasm. PMID- 10818334 TI - Methemoglobinemia after topical anesthesia with lidocaine and benzocaine for a difficult intubation. AB - Methemoglobinemia is an uncommon cause of cyanosis; however, rapid diagnosis is critical to avoid potentially fatal consequences. Several local anesthetics can precipitate methemoglobinemia in susceptible patients. This case report of acute methemoglobinemia occurred during fiberoptic intubation in an awake patient without a past medical history of adverse reactions to local anesthetics. PMID- 10818335 TI - Comparative study of Mm. Multifidi in lumbar and thoracic spine. AB - Imbalance of Mm. Multifidi may play a role in spinal disorders such as scoliosis in the thoracic spine, and lumbar disc herniation and lower back pain in the lumbar spine. Even though changes in these muscles are related to the etiology of these disorders, their anatomy is still poorly understood, especially in the upper regions of the spine. With the aim of gaining a better understanding of the anatomy of Mm. Multifidi in the lumbar and thoracic spine, 12 fresh and two embalmed cadavers were dissected. Our results indicate that Mm. Multifidi present differences in lumbar and thoracic spines concerning their deepness, fibre trajectory, muscle length, muscle mass and tendinous tissue. In the lumbar spine Mm. Multifidi are a superficial, thick and fleshy mass, and their fibres are more vertical in relation to the spinous processes. In the thoracic spine Mm. Multifidi are deeper, thinner, and their fibres are more tendinous and oblique than in the lumbar spine. These differences have implications on Mm. Multifidi architecture and consequently for their function in these two regions of the spine. PMID- 10818336 TI - Between-days reliability of subjective and objective assessments of back extensor muscle fatigue in subjects without lower-back pain. AB - BACKGROUND: It is important to evaluate the reliability of common used methods of examining muscle fatigue from the lower back since the methods are used in patient evaluation. METHODS: To establish between-days reliability, ten subjects without lower-back pain performed a Sorensen test, a prone test for back extensor muscles against gravity, on three separate days. EMG was recorded from the L1 and L5 of the back extensor muscles. Fatigue was subjectively rated using a Borg CR 10 scale. Intraclass correlation coefficient, standard error of measurement and coefficient of variation were calculated from a one-way ANOVA. Percent agreement was also calculated. RESULTS: The study revealed good reliability for the slope for the total time (ICC 0.65-0.90), the initial and end median frequency (ICC 0.75-0.89), median frequencies at Borg ratings of three (ICC 0.63-0.88), five (ICC 0.62-0.84) and seven (ICC 0.67-0. 87), endurance time (ICC 0.89). The Borg ratings of the first minute agreed better than those of the second and the third. The Borg ratings at the second and the third test agreed to 40-80%, indicating a need for a practice session. CONCLUSION: The protocol used for assessing fatigue in the back extensor muscles proved to be reliable and is recommended for further use. PMID- 10818337 TI - Assessment of neuromuscular response characteristics at the knee following a functional perturbation. AB - Previous research indicates that both the extent and timing of muscular activation at the knee can be influenced by muscle activity state, joint angle, weight-bearing status and trunk position. However, little research to date has evaluated protective neuromuscular response times and activation patterns to an imposed perturbation with the knee joint in a functional, weight-bearing stance. Hence, we designed a lower extremity perturbation device to produce a sudden, forward and either internal or external rotation moment of the trunk and femur relative to the weight-bearing tibia. Surface electromyography (EMG) recorded long latency reflex times of the medial and lateral quadriceps, hamstring and gastrocnemius muscles in 64 intercollegiate lacrosse and soccer players in response to both internal and external rotation perturbation. We found the gastrocnemius fired significantly faster that the hamstring, which in turn fired significantly faster than the quadriceps. There was also a significant difference in activation times of the medial and lateral hamstring not found for the quadriceps or gastrocnemius muscles. Our findings confirmed that reactive neuromuscular responses following this functional perturbation differ markedly from those previously reported using seated, partial weight-bearing and/or uni planar models under relaxed conditions. PMID- 10818338 TI - Anticipatory postural adjustments depend on final equilibrium and task complexity in vertical high jump movements. AB - Anticipatory postural adjustments (APA) associated with high vertical jumping were studied in five normal men. The dependence of APA on final equilibrium and task complexity was studied by analyzing the characteristics of single versus repetitive jumps. Subjects performed series of 25 single jumps and 25 repetitive jumps on a force platform, in response to a Go signal. Surface EMG from the Soleus (SO), Tibialis anterior (TA), Vastus lateralis (VL) and Biceps femoris (BF) muscles were recorded simultaneously with the three components of the ground reaction forces. The existence of APA associated with both single and repetitive jumps was evidenced by a backward shift of the center of pressure, an SO deactivation, and a TA activation which preceded the onset of movement by hundreds of milliseconds. In repetitive jumps, SO deactivation occurred later than in single jump, whereas the center of pressure shift started earlier, lasted longer and had a greater amplitude. Moreover, repetitive jumps were initiated later with respect to the Go signal. These data show that the central nervous system anticipates not only the equilibrium disturbance caused by an initial movement but that caused by a whole sequence of movements. They also show that the corresponding APA are based heavily on a single functional ankle synergy. PMID- 10818339 TI - Reflex facilitation during the stretch-shortening cycle. AB - Maximal torque during the concentric phase of a movement has been shown to be enhanced by prior eccentric muscle actions, a movement strategy referred to as the stretch-shortening cycle. Although the mechanical basis for this enhancement is well established, the neural component is not. We hypothesized that brief high frequency bursts of spindle afferent discharge during the eccentric phase of the stretch-shortening cycle could be one mechanism for facilitating the volitional drive. To test this hypothesis, three sets of experiments were done. In the first (N=15), we demonstrated that both the peak and mean EMG of the soleus (S) and lateral gastrocnemius (LG) muscles were considerably greater during a reciprocal hopping (RHOP) task than for maximum isometric contractions (MIVCs). In the second experiment, we tested whether the dynamic nature of the RHOP or the eccentric phase of the RHOP contributed to the EMG potentiation. Peak and mean EMG produced with a concentric hop (CHOP), in which the lengthening phase of the hop was eliminated, were compared with that produced with the RHOP and MIVCs conditions (N=7). The RHOP produced greater peak EMG than either the CHOP or the MIVCs while the mean EMG for both hopping conditions was considerably more than the MIVCs. In the final experiment, we attempted to mimic the brief high frequency burst of spindle afferent activity during the lengthening phase of the stretch-shortening cycle in the absence of muscle length changes. High-frequency (100 Hz) afferent stimulation (HFS) was delivered during MIVCs. At rest, the HFS produced negligible EMG activity but when superimposed over MIVCs produced a marked potentiation of the S EMG over values obtained during MIVCs alone. Evidence that HFS synchronizes the EMG associated with volitional activation is also provided. We conclude that a substantial but brief facilitation and possible synchronization of the neural drive is provided by the spindle afferents during the eccentric phase of the stretch-shortening cycle. PMID- 10818340 TI - Gender and muscle differences in EMG amplitude and median frequency, and variability during maximal voluntary contractions of the quadriceps femoris. AB - The purpose of this study was to evaluate gender and muscle differences in electromyographic (EMG) amplitude and median frequency mean and standard deviation during maximal voluntary contractions of the quadriceps femoris. Thirty recreationally active volunteers were assessed for isometric EMG activity of the vastus medialis (VM), vastus lateralis (VL), and rectus femoris (RF) muscles during three 5-s maximal isometric voluntary contractions (MVCs). Median frequency of the three muscles was assessed through a power spectral analysis (fast Fourier transformation, Hanning window processing, 512 points). The power spectral analysis was performed during the middle 3 s of each contraction over 11 consecutive, 512 ms epochs overlapping each other by half their length (256 ms). The median frequency (F(med)) for each of the 11 windows was determined for each muscle. The mean and standard deviation of the F(med) across the 11 overlapping windows were then calculated for each contraction and muscle. EMG amplitude was determined by calculating the root mean square (RMS-50 ms time constant) over the same contraction period for each muscle. The mean amplitude and standard deviation about the mean value were then determined. A three-factor ANOVA with repeated measures was performed on the calculated F(med) mean and standard deviation values, and RMS standard deviations, to assess any gender, muscle, or trial differences, or interactions. A two-factor (gender by muscle) ANOVA with repeated measures was performed on the RMS mean amplitude for each muscle. Intraclass correlation coefficients (ICCs-2,1), standard errors of measurement (SEMs), and associated 95% confidence intervals were then calculated for maximal quadriceps torque and F(med) for each muscle. The results from this study demonstrated that the VL muscle displayed significantly higher F(med) values than the RF and VM muscles. The RF muscle showed significantly higher F(med) values (mean of 11 overlapping windows) than the VM muscle. Intrasession reliability was found to be high for the calculated mean values (ICC=0.85-0.96), but was shown to be low for variability (ICC=0.13-0.45). The major findings of this study support the notion that the EMG signal is "quasi-random" in nature, as demonstrated by the reproducible F(med) means and unreliable variability. PMID- 10818341 TI - EMG and kinematics of normal subjects performing trunk flexion/extensions freely in space. AB - Ten normal subjects performed continuous trunk flexion/extensions (F/E) without any restraining apparatus at free, 3, 2.25 and 1.5 s periods and a fatiguing task of F/E at 1.5 s period during 45 s. Kinematics of the trunk was obtained with bilateral electromyographic (EMG) activity of the erector spinae (three levels), the abdominal oblique muscles and the rectus abdominis muscles. The free period chosen by the subjects was found to vary between 3.05 and 1.47 s. Lateral flexion of the spine was similar in each task but rotation about its longitudinal axis increased as the F/E period shortened. When left and right side EMG signals were grouped by level of recording, a significant difference in activity was found. Subjects who produced the slowest free F/E displayed larger fatigue indexes derived from the EMG signals for some of their back muscles than for other subjects. The flexion/relaxation phenomenon was considered present in a muscle if a level <10% of the maximum signal recorded during extension was detected. The phenomenon was found in >50% of the observations and occurred at a similar angle in each task. Kinematics and several characteristics of the EMG signals of the trunk were statistically independent of the speed of motion. PMID- 10818342 TI - Upper limb H reflexes and somatosensory evoked potentials modulated by movement. AB - In the human lower limb, the magnitudes of both Hoffmann (H) reflexes and primary somatosensory evoked potentials (SEPs) from scalp electrodes, are reduced by active and/or passive movement. We surmised that similar effects occur for the upper limb and specifically hypothesised that amplitudes of median nerve induced flexor carpii radialis H reflexes and cortical SEPs are reduced with passive movement about the wrist or elbow. The results showed (P<0. 05) that either movement significantly attenuated mean magnitudes of SEPs elicited from stimulation at elbow or wrist and that reflex magnitudes attenuated with wrist movement. Thus, the upper limb shows similar movement-induced modulation to the lower limb. These attenuations of fast conducting sensory paths consequent to movement per se, may be a basic level of motor control, initiated from muscle mechanoreceptor discharge. Upon this basic level, more complex modulations then may be laid as appropriate for the particular characteristics of active motor tasks. PMID- 10818343 TI - The 1.30 A resolution structure of the Bacillus subtilis chorismate mutase catalytic homotrimer. AB - The crystal structure of the Bacillus subtilis chorismate mutase, an enzyme of the aromatic amino acids biosynthetic pathway, was determined to 1.30 A resolution. The structure of the homotrimer was determined by molecular replacement using orthorhombic crystals of space group P2(1)2(1)2(1) with unit cell parameters a = 52.2, b = 83. 8, c = 86.0 A. The ABC trimer of the monoclinic crystal structure [Chook et al. (1994), J. Mol. Biol. 240, 476-500] was used as the starting model. The final coordinates are composed of three complete polypeptide chains of 127 amino-acid residues. In addition, there are nine sulfate ions, five glycerol molecules and 424 water molecules clearly visible in the structure. This structure was refined with aniosotropic temperature factors, has excellent geometry and a crystallographic R factor of 0.169 with an R(free) of 0.236. The three active sites of the macromolecule are at the subunit interfaces, with residues from two subunits contributing to each site. This orthorhombic crystal form was grown using ammonium sulfate as the precipitant; glycerol was used as a cryoprotectant during data collection. A glycerol molecule and sulfate ion in each of the active sites was found mimicking a transition state analog. In this structure, the C-terminal tails of the subunits of the trimer are hydrogen bonded to residues of the active site of neighboring trimers in the crystal and thus cross-link the molecules in the crystal lattice. PMID- 10818344 TI - Structure of buffalo lactoferrin at 3.3 A resolution at 277 K. AB - The three-dimensional structure of diferric buffalo lactoferrin has been determined at 3.3 A resolution. The structure was solved by molecular replacement using the coordinates of diferric human lactoferrin as a search model and was refined by simulated annealing (X-PLOR). The final model comprises 5316 protein atoms for all 689 residues, two Fe(3+) and two CO(3)(2-) ions. The final R factor was 21.8% for 11 711 reflections in the resolution range 17.0-3.3 A. The folding of buffalo lactoferrin is essentially similar to that of the other members of the transferrin family. The significant differences are found in the dimensions of the binding cleft and the interlobe orientation. The interlobe interactions are predominantly hydrophobic in nature, thus facilitating the sliding of two lobes owing to external forces. The interdomain interactions are comparable in the N and C lobes. PMID- 10818345 TI - Structures of oxidized and reduced azurin II from Alcaligenes xylosoxidans at 1.75 A resolution. AB - Crystallographic structures of oxidized and reduced forms of azurin II are reported at 1.75 A resolution. Data were collected using one crystal in each case and by translating the crystal after each oscillation range to minimize photoreduction. Very small differences are observed at the Cu site upon reduction and these cannot be determined with confidence at current resolution. A comparison with the three-dimensional EXAFS reveals a good correspondence for all the ligand distances except for Cu-His46, where a larger deviation of approximately 0.12-0.18 A is observed, indicating that this ligand is more tightly restrained in the crystallographic refinement at the current resolution. PMID- 10818346 TI - 3D EXAFS refinement of the Cu site of azurin sheds light on the nature of structural change at the metal centre in an oxidation-reduction process: an integrated approach combining EXAFS and crystallography. AB - Three-dimensional information is obtained for the Cu site in azurin at very high resolution by combining high-resolution crystallographic structures and EXAFS data for the oxidized and reduced form of the protein. This combined approach has allowed us to define the subtle structural changes (<0.1 A) which take place at the Cu site during a single-electron redox process. PMID- 10818347 TI - Ab initio structure determination of the lantibiotic mersacidin. AB - The crystal structure of mersacidin, a potential novel antibiotic against methicillin- and vancomycin-resistant Staphylococcus aureus strains, has been determined by ab initio methods. Despite all crystals being merohedrally twinned, an accurate structural model with an R value of 13.4% has been obtained at atomic resolution. With six molecules in the asymmetric unit and no atom heavier than sulfur, the structure corresponds to a protein of 120 amino acids and is the largest approximately equal-atom unknown structure solved by direct methods. In the crystal, the molecule assumes a compact fold different from that found by NMR in solution. Comparison of the NCS-related molecules reveals regions of variable flexibility. The region highly homologous to the related antibiotic actagardine is very rigid and possibly defines an essential building block of this class of new antibacterial substances. PMID- 10818348 TI - Objective comparison of protein structures: error-scaled difference distance matrices. AB - Understanding of macromolecular function in many cases relies on the comparison of related structural models. Commonly used least-squares superposition methods suffer from bias introduced into the comparison process by the subjective choice of atoms employed for the superposition. Difference distance matrices are a more objective means of comparing structures as they do not depend on a particular superposition scheme. However, they suffer from very high noise originating from coordinate errors. Modern refinement programs allow the rigorous estimation of standard uncertainties for individual atomic positions. These errors can be propagated through the calculation of a difference distance matrix allowing one to assess the significance level of structural differences. An algorithm is presented which produces an intuitive graphical representation of difference distance matrices after normalization to their error levels. Two examples where its application was revealing are described. Alternatives are suggested for cases where rigorous estimation of individual errors by the inversion of the full least squares matrix is not feasible. The method offers an unbiased way to detect significant similarities and differences between related structures, as encountered in studies of complexes and mutants or when multiple models are obtained from experiments such as crystal structures involving non crystallographic symmetry or different crystal modifications, or ensembles derived from NMR spectroscopy. PMID- 10818349 TI - Determining protein structure from electron-density maps using pattern matching. AB - TEXTAL is an automated system for building protein structures from electron density maps. It uses pattern recognition to select regions in a database of previously determined structures that are similar to regions in a map of unknown structure. Rotation-invariant numerical values, called features, of the electron density are extracted from spherical regions in an unknown map and compared with features extracted around regions in maps generated from a database of known structures. Those regions in the database that match best provide the local coordinates of atoms and these are accumulated to form a model of the unknown structure. Similarity between the regions in the database and an uninterpreted region is determined firstly by evaluating the numerical difference in feature values and secondly by calculating the electron-density correlation coefficient for those regions with similar feature values. TEXTAL has been successful at building protein structures for a wide range of test electron-density maps and can automatically model entire protein structures in a few hours on a workstation. Models built by TEXTAL from test electron-density maps of known protein structures were accurate to within 0.6-0.7 A root-mean-square deviation, assuming prior knowledge of C(alpha) positions. The system represents a new approach to protein structure determination and has the potential to greatly reduce the time required to interpret electron-density maps in order to build accurate protein models. PMID- 10818350 TI - A new vector-search rotation function: image-seeking functions revisited in macromolecular crystallography. AB - A new rotation function in Patterson space is described. An image-seeking function can be defined as a criterion of fit between the observed Patterson map and a suitable vector set extracted from a specially calculated Patterson map of the search model. The behaviour of image-seeking functions has appeared to be heavily dependent on certain relations between some statistical parameters of both maps. A new algorithm, which carries out the crucial step of selecting the appropriate vector set from the search model, has been established. As a consequence of the combination of these two preceding results, a new vector search rotation function has been proposed and tested. PMID- 10818351 TI - Crystallization of Escherichia coli beta-ketoacyl-ACP synthase III and the use of a dry flash-cooling technique for data collection. AB - beta-Ketoacyl-acyl carrier protein (ACP) synthase III (FabH) is a condensing enzyme active in the fatty-acid biosynthesis pathway of bacteria. The enzymes of this pathway provide a set of targets for the discovery of previously unknown antibiotics. FabH from Escherichia coli has been crystallized in two crystal forms using the sitting-drop vapor-diffusion technique. The first form crystallized in the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 63.1, b = 65.1, c = 166.5 A; the second form crystallized in the tetragonal space group P4(1)2(1)2, with unit-cell parameters a = b = 72.7, c = 99.8 A. A flash-cooling technique using no cryoprotectant was utilized in obtaining data from the second type of crystals. PMID- 10818352 TI - Crystallization and preliminary X-ray analysis of high-alkaline pectate lyase. AB - Pel-15, a high-alkaline pectate lyase (pectate transeliminase; E.C. 4.2.2.2) from Bacillus sp. strain KSM-P15, has been crystallized using the hanging-drop vapour diffusion method at 277 K. Two different crystal forms were obtained and preliminary X-ray diffraction data were collected from each crystal form at 100 K. Both forms belong to the orthorhombic space group P2(1)2(1)2(1) and contain one molecule per asymmetric unit. The unit-cell parameters of form I are a = 43.2 (2), b = 60.2 (2), c = 82.2 (2) A and those of form II are a = 42.9 (1), b = 43.4 (1), c = 105.9 (3) A. Diffraction data to a resolution of 1.5 A were collected from form II crystals using a synchrotron-radiation source. PMID- 10818353 TI - Purification, crystallization and preliminary X-ray analysis of a complex between granulocyte colony-stimulating factor and its soluble receptor. AB - Crystals of the complex between granulocyte colony-stimulating factor and its soluble receptor were obtained by a vapour-diffusion method using ammonium sulfate as a precipitant. Addition of 1, 4-dioxane was critical in order to grow the crystals to sufficient sizes. Cryoprotection was essential in order to collect diffraction data at atomic resolution. Two kinds of crystal forms were obtained depending on the cryoprotectants. In a cryosolvent with the same salt concentration as in the crystallization conditions, the crystal belonged to the space group I4(1)22. At higher salt concentrations, the crystal was converted to a different space group P4(1)2(1)2 (P4(3)2(1)2) with the same unit-cell parameters. PMID- 10818354 TI - Preliminary X-ray crystallographic study of DsrD protein from the sulfate reducing bacterium Desulfovibrio vulgaris. AB - DsrD (dissimilatory sulfite reductase D) protein encoded by the dsr operon of the sulfate-reducing bacterium Desulfovibrio vulgaris Hildenborough has been crystallized using the vapour-diffusion method with ammonium sulfate as a precipitating agent. The crystals diffract to 1.7 A resolution and belong to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 60.54 (6), b = 65. 20 (4), c = 46.41 (3) A. The crystal contains two DsrD molecules per asymmetric unit, giving a Matthews coefficient (V(M)) of 2.6 A(3) Da(-1). A gold derivative (NaAuCl(4)) crystal has been successfully prepared. PMID- 10818355 TI - Crystallization and preliminary X-ray crystallographic analysis of thioesterase I from Escherichia coli. AB - The Escherichia coli thioesterase I specifically catalyzes the deacylation of fatty acyl-CoA thioesters, especially those with long acyl groups (C(12)-C(18)). Single crystals of thioesterase I (E.C. 3. 1.2.2) from E. coli have been obtained using methoxypolyethylene glycol 5000 (PEG-MME 5K) as a precipitant at room temperature in 21 d. The crystals belong to the tetragonal space group P4(1)2(1)2 or its enantiomorph P4(3)2(1)2, with unit-cell parameters a = b = 50.85 (7), c = 171.5 (1) A. The crystals diffract to beyond 2.4 A resolution. There is one molecule of molecular weight 20.5 kDa in the asymmetric unit, with a solvent content of 55%. PMID- 10818356 TI - Crystallization and preliminary X-ray analysis of the matrix protein from Ebola virus. AB - The matrix protein from Ebola virus is a membrane-associated molecule that plays a role in viral budding. Despite its functional similarity to other viral matrix proteins, it displays no sequence similarity and hence may have a distinct fold. X-ray diffraction quality crystals of the Ebola VP40 matrix protein were grown by the hanging-drop vapour-diffusion method. The crystals belong to the monoclinic space group C2, with unit-cell parameters a = 64.4, b = 91.1, c = 47.9 A, beta = 96.3 degrees. A data set to 1.9 A resolution has been collected using synchrotron radiation. The unit cell contains one molecule of molecular weight 35 kDa per asymmetric unit, with a corresponding volume solvent content of 35%. PMID- 10818357 TI - Crystallization and initial crystallographic analysis of phosphomannomutase/phosphoglucomutase from Pseudomonas aeruginosa. AB - The enzyme phosphomannomutase/phosphoglucomutase (PMM/PGM) catalyzes the conversion of mannose 6-phosphate to mannose 1-phosphate in the second step of the alginate biosynthetic pathway of Pseudomonas aeruginosa. PMM/PGM has been crystallized by hanging-drop vapor diffusion in space group P2(1)2(1)2(1). Crystals diffract to 1.75 A resolution on a synchrotron X-ray source under cryo cooling conditions. PMM/PGM substituted with selenomethionine has been purified and crystallizes isomorphously with the native enzyme. Structure determination by MAD phasing is under way. Because of its role in alginate biosynthesis, PMM/PGM is a potential target for therapeutic inhibitors to combat P. aeruginosa infections. PMID- 10818358 TI - Crystallization and aldo-keto reductase activity of Gcy1p from Saccharomyces cerevisiae. AB - Crystallization and preliminary X-ray diffraction studies of Gcy1p, an aldo-keto reductase from Saccharomyces cerevisiae, have been performed. Both the wild type and a double-mutant form of Gcy1p were crystallized using the hanging-drop method at 298 K; however, only the double-mutant form has so far yielded crystals suitable for X-ray diffraction analysis. These crystals belonged to the primitive monoclinic space group P2(1), with unit-cell parameters a = 50.94, b = 65.64, c = 86.23 A, beta = 92.64 degrees. Diffraction data were collected to 2.5 A. Assuming two 35 kDa subunits in the asymmetric unit yielded a V(m) of 2.06 A(3) Da(-1). Additionally, a kinetic study performed by measuring the rate of oxidation of NADPH in the presence of several substrates indicates that both wild-type and double-mutant proteins are enzymes possessing NADPH-dependent reductase activity. PMID- 10818359 TI - Expression, purification, crystallization and preliminary X-ray analysis of the iota-carrageenase from Alteromonas fortis. AB - This is the first crystallization report of a glycoside hydrolase which belongs to family 82. A recombinant form of His-tagged iota-carrageenase from Alteromonas fortis was expressed, purified and crystallized. Crystals were obtained by the vapour-diffusion method using polyethylene glycol (M(W) = 6000) as a precipitant. They belong to space group P2(1), with unit-cell parameters a = 56. 75, b = 91.04, c = 125.01 A, beta = 93.41 degrees. The unit cell contains two molecules in the asymmetric unit related by a non-crystallographic twofold axis. Crystals diffracted to 2.0 A resolution on a synchrotron beamline. PMID- 10818360 TI - Crystallization and preliminary crystallographic studies of a SAM domain at the C terminus of human p73alpha. AB - p73 is a recently discovered homologue of the tumour suppressor p53 and contains all three functional domains of p53. The alpha-splice variant of p73 (p73alpha) contains an additional structural domain near its C--terminus that has sequence homology with the sterile alpha-motif (SAM) domain. This domain is considered to be responsible for mediating protein-protein interactions. Pyramidal crystals of human p73alpha SAM domain were obtained by the hanging-drop vapour-diffusion method with ammonium dihydrogen orthophosphate as the precipitant. The crystals diffract to 2.54 A resolution and belong to the tetragonal space group P4(1)2(1)2, with unit-cell parameters a = b = 32.02, c = 133.84 A. The structure was solved by molecular replacement using the NMR structure of the same protein as the search model. PMID- 10818361 TI - Crystallization and preliminary X-ray analysis of human grancalcin, a novel cytosolic Ca2+-binding protein present in leukocytes. AB - Recombinant human grancalcin, a calcium-binding protein from leukocytes, has been crystallized in the presence or absence of Ca(2+) by the vapor-diffusion method. Two crystal forms of apo grancalcin were obtained: space group P2(1), with unit cell parameters a = 48.4, b = 81.1, c = 46.6 A, beta = 111.3 degrees, diffracting to 1.9 A, and space group C2, with unit-cell parameters a = 97.0, b = 51.9, c = 75.9 A, beta = 108.5 degrees, diffracting to 2.4 A. Crystals were also grown in the presence of 5 mM Ca(2+). They also belong to space group C2, with unit-cell parameters a = 97.4, b = 50.3, c = 77.6 A, beta = 108.2 degrees, which are very similar to the second apo grancalcin form. These crystals diffract to 2.5 A. PMID- 10818362 TI - Overexpression, crystallization and preliminary X-ray crystallographic analysis of dihydrofolate reductase from bacteriophage T4. AB - Dihydrofolate reductase (DHFR) from bacteriophage T4 is a homodimer consisting of 193-residue subunits. It has been crystallized in the presence of the cofactor (NADPH) and an inhibitor (aminopterin) at 296 K using sodium chloride as precipitant. The crystals are tetragonal, belonging to the space group P4(1)22 (or P4(3)22), with unit-cell parameters a = b = 61.14, c = 123.23 A under cryogenic conditions. The asymmetric unit contains a single subunit, with a corresponding V(m) of 2.65 A(3) Da(-1) and a solvent content of 53. 6%. Native data have been collected from a crystal to 1.9 A resolution using synchrotron X rays. PMID- 10818363 TI - Crystallization and preliminary X-ray diffraction analysis of Saccharomyces cerevisiae Ygr203p, a homologue of Acr2 arsenate reductase. AB - Ygr203p, a 148-residue protein encoded by the ygr203w gene of Saccharomyces cerevisiae, is a homologue of the yeast Acr2 arsenate reductase encoded by the acr2 (or ypr200c) gene. It also shows significant sequence similarity to the human cell-cycle control Cdc25 phosphatase family. It has been overexpressed in soluble form in Escherichia coli with a His(6) tag at its C-terminus. The recombinant protein has been crystallized at 296 K using sodium chloride as precipitant. The crystals belong to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 40.48, b = 50.95, c = 91.95 A. The asymmetric unit contains a monomer, giving a crystal volume per protein mass (V(m)) of 2.61 A(3) Da(-1) and a solvent content of 53.8%. The crystals diffract to better than 1.9 A resolution with Cu Kalpha X-rays. They are therefore suitable for high-resolution structure determination. PMID- 10818364 TI - Crystallization in cubo: general applicability to membrane proteins. AB - Obtaining well ordered crystals of membrane proteins is the single most serious stumbling block in the pursuit of their high-resolution structures. The applicability of lipidic cubic phase-mediated crystallization is demonstrated on a diverse set of bacterial membrane proteins: two photosynthetic reaction centres, a light-harvesting complex and two retinal proteins, halorhodopsin and bacteriorhodopsin. Despite marked differences in molecular dimensions, subunit composition and membrane origin, one single lipid, monoolein, is sufficient to form a crystallization matrix for all the aforementioned systems. Therefore, the lipidic cubic phase approach is proposed as a general method for crystallizing membrane proteins. PMID- 10818365 TI - Oxidation of selenomethionine: some MADness in the method! AB - Since it was first reported, the multiwavelength anomalous diffraction (MAD) technique for the determination of protein structures has become widely accepted and increasingly popular. Here, it is demonstrated that the anomalous signal from selenomethione (SeMet) substituted proteins can be significantly enhanced by oxidation. PMID- 10818368 TI - Endless comparisons. PMID- 10818369 TI - It's a knockout: survey of the management of avulsed teeth. AB - Avulsion of a tooth is a not uncommon childhood injury, with up to 16% of dental injuries involving the loss of a tooth. Although this is an apparently 'minor' injury, for a child there may be long-term psychological and treatment problems. Correct advice in the early stages of this injury, and the recognition that prompt treatment may allow a tooth to be successfully reimplanted are vital in the management of these injuries. Often, nursing staff in the accident and emergency or paediatric department may be asked for advice either professionally or informally, and can influence the long-term dental health for these patients. This paper looks at the knowledge of parents and healthcare professionals in the management of these injuries. PMID- 10818370 TI - A study of the effects of the appointment of a clinical nurse educator in one Victorian emergency department. AB - The Clinical Nurse Educator (CNE) project saw the appointment and evaluation of a CNE position in the Emergency Department at Dandenong Hospital, Australia. The study aimed to identify the educational needs of nursing staff, the self reported levels of knowledge of nursing staff, the perceptions of nursing staff surrounding education and clinical support and to compare responses over the 6 month period to identify any statistically significant changes. Data was collected at three intervals during the study period and the responses compared using the Kruskal-Wallis test(H). Since the appointment of the CNEs, the reported levels of knowledge increased for all areas of emergency nursing included in the study. Tutorials, in-service education sessions, direct clinical support and self directed learning packages were reported to be useful educational methods. There were increases in the reported adequacy of in-service education (P = 0.0000), level of clinical support and satisfaction with current level of knowledge in emergency nursing. Nursing staff found the process of basic and advanced life support assessment less intimidating (P = 0.0031), more important and less affected by workload constraints of the ED (P = 0.0002). The reported thoroughness of orientation of new employees (P = 0.0005) and levels of clinical support and education when orientated to the triage role (P = 0.0225) also increased. PMID- 10818371 TI - Dilemma. Role conflict. PMID- 10818372 TI - Closer professional links--a must for the future NHS. AB - Inter-professional collaboration between doctors, nurses and the professions allied to medicine (PAMs) will have to increase if the quality of patient care is to be sustained and improved, according to researchers based at Sheffield University's School of Health and Related Research (ScHARR). They have been collaborating with teams at the University of Bristol's School for Policy Studies, and the King's Fund Nursing Developments Programme. PMID- 10818373 TI - Head lice for A & E nurses. AB - Accident and emergency nurses are in an excellent position to offer skill and advice on the treatment of head lice, from health education to diagnosis and cure to contact tracing. This paper offers guidance on how to detect lice and how to treat infestation. There are a number of treatment options available, including pharmacological and physical treatments. Choosing the most appropriate option can be difficult due to reports of 'Super Nits' resistant to insecticides and lack of data to support the use of alternative methods such as 'wet combing' and aromatherapy treatments. However, nurses do need to be aware of how lice move between people, so they can take an active role in health education and make sure that patients affected by head lice are managed properly. PMID- 10818374 TI - Student approaches: supporting students in A&E settings. AB - Nurses working in accident and emergency (A&E) departments near to Higher Education Institutions will be familiar with students attending as the result of sports injuries, and as the consequence of a range of accidents stemming from over-enthusiastic socializing. However, a number of students arrive at A&E departments in distress as the result of mental health problems. This article reports the findings of a small research project exploring mechanisms and support systems for assisting students with mental health needs. It briefly outlines the research and its findings and then considers three relevant issues for those working in A&E: student profiles, student support and the position of students who are considering or engaged in professional education. The article ends with a brief discussion of the position of students undertaking placements within A&E departments. PMID- 10818375 TI - Children's nurses in the accident and emergency department: literature review. AB - It is estimated that over 30% of Accident and Emergency attendees are children; despite this, many hospitals still do not have arrangements for them. Children often receive care from health professionals who are unaware of their specialist requirements. If we are to meet the demands of the document A First Class Service (National Health Service Executive 1998), which states that all patients are entitled to high quality care, emergency services for children and young people must be planned specifically, as an adjunct to adult services. PMID- 10818377 TI - An evaluation of the care of patients with minor injuries in emergency settings. AB - This paper describes a study which evaluated three models of emergency care for patients with minor injuries; a traditional Accident and Emergency (A&E) department where patients with minor injuries were seen by senior house officers and nurses working together; a Minor Accident Treatment Service (MATS) unit, located in a main A&E, where patients with minor injuries were seen by Emergency Nurse Practitioners (ENPs) and a nurse-led Minor Injuries Unit (MIU), where patients with minor injuries were also seen by ENPs. Information about waiting times and length of stay was recorded by the researchers and patients were asked to complete a satisfaction questionnaire at the end of their stay. A sample of 181 patients were included in the study (57 in the traditional A&E, 57 in the MATS unit and 67 in the nurse-led MIU). Patients in the traditional A&E waited significantly longer to be seen than patients seen by an ENP, by an average of 40 minutes, and spent an average of 33 minutes longer in the department. ENPs spent an average of 12 minutes longer than doctors undertaking the first consultation where patients' needs were assessed and documented. Patients were very satisfied with care given by all health professionals. Those seen by ENPs were significantly more likely to have been given health advice and information, written instructions on discharge and told who to contact if they needed further advice. They were found to be subsequently significantly less worried about their health than patients seen in the traditional A&E. It is suggested that although ENPs spent longer with patients, this resulted in an improvement in the quality and depth of information and advice provided. PMID- 10818378 TI - Nurse practitioners and the Ottawa Ankle Rules: comparisons with medical staff in requesting X-rays for ankle injured patients. AB - This paper reports on part of a study of 354 consecutive attendances to an Accident and Emergency (A&E) department by patients with ankle injuries. The aim of this study was to examine the percentage of X-ray photographs requested by nurse practitioners compared to those requested by medical practitioners. The study took place in an A&E department where nurse practitioners have the authority to request X-rays photographs for ankle injuries, either at triage assessment or at the subsequent treatment stage. Nurse practitioners applied the Ottawa Ankle Rules (OAR) in 187 patients. The results show that nurse practitioners requested X-rays for 61.5% of patients assessed using the OAR. This is compared with 80.4% for patients assessed by medical practitioners. The difference is highly significant. Detection rates show that fractures were identified in 29.6% of patients sent for X-ray by nurse practitioners compared with 22.8% in patients seen by medical practitioners, although this difference is not significant. Although nurse practitioners applied the OAR appropriately in all 187 cases, four patients who were assessed by nurse practitioners and judged not to need an X-ray photograph were subsequently found to have a fracture. PMID- 10818379 TI - Headaches: how serious are they and when do you admit? PMID- 10818382 TI - The impact of the quality agenda on workforce planning. PMID- 10818383 TI - Taking the advanced knee arthroscopic surgery course to Singapore. AB - The Advanced Knee Arthroscopic Surgery course was first organised in 1995 under the joint auspices of the British Association for Surgery of the Knee and The Royal College of Surgeons of England. This is a two-day course in advanced knee arthroscopic techniques, with lectures, videos and extensive practical sessions. The approach is very much hands-on, using cadavers together with sawbones and model knees. Use is also made of the specimens in the Anatomical Museum of the College. Instruction is given in anterior cruciate ligament reconstruction, meniscal repair and posterior cruciate with posterolateral ligament reconstruction. Since 1995 the course has been held on seven occasions and has been attended by over 100 senior trainees and consultants. PMID- 10818384 TI - Core skills, courses and competency. PMID- 10818385 TI - Introduction: setting reproductive health priorities to meet the needs of the new millennium. PMID- 10818386 TI - Neuropeptides, neurotransmitters, neurosteroids, and the onset of puberty. AB - Puberty results from withdrawal of the "gonadostat" mechanisms and from increased gonadotropin sensitivity to GnRH. It has been hypothesized that GnRH release may be modulated by a non-steroid-mediated mechanism. Modifications of neuropeptides, neurotransmitters, and neurosteroids may underlie the onset of pubertal processes. Neuropeptides mainly involved in the control of GnRH release are opioids, neuropeptide Y (NPY), galanin, and corticotropin-releasing factor (CRF), whereas neurotransmitters are noradrenaline, dopamine, serotonin, melatonin and gamma-aminobutyric acid (GABA). Norepinephrine, epinephrine, and dopamine stimulate GnRH, whereas the effect of serotonin on hypothalamic-pituitary-ovarian axis seems to be norepinephrine-mediated. Neurosteroids are steroid hormones that bind to the GABA-A receptor, synthesized in the brain de novo or from blood-borne precursors. DHEA, a GABA-A antagonistic neurosteroid, and allopregnanolone, a GABA-A agonistic neurosteroid, may be important in the onset of gonadarche. In conclusion, the onset of puberty derives from the complex interplay among neuropeptides, neurotransmitters, and neurosteroids that occurs in the awakening of hypothalamic-pituitary-ovarian axis. PMID- 10818387 TI - Ovarian regulators of gonadotropin secretion. AB - Great progress in the ovarian mechanisms that control gonadotrophin secretion in women has recently been achieved. Estradiol (E2) is the main component of the ovarian negative effect on basal gonadotropin secretion during the normal menstrual cycle. However, nonsteroidal substances such as inhibins and activins that can affect follicle-stimulating hormone (FSH) secretion in vitro may also participate in the control of FSH secretion in vivo. Recent evidence has shown that the ovaries also produce another nonsteroidal substance, named gonadotropin surge-attenuating factor (GnSAF), that specifically attenuates GnRH-induced LH secretion and the endogenous LH surge in superovulated women. It is possible that during the normal menstrual cycle GnSAF controls the amplitude of the midcycle LH surge by antagonizing the stimulating effect of E2 on the pituitary. PMID- 10818389 TI - A clinical understanding of the estrogen receptor. PMID- 10818388 TI - Progesterone is an autocrine/paracrine regulator of human granulosa cell survival in vitro. AB - Ovarian follicles are composed of granulosa cells (GC), which undergo apoptosis within 24 hours of culture in serum-free medium. The present study was designed to assess the role of progesterone in regulating human GC survival. Human GC were isolated from follicular aspirates of women undergoing in vitro fertilization. GC were then cultured for 24 hours in serum-free media supplemented with progesterone and/or the progesterone antagonist RU486 and dexamethasone. Cells were then fixed and assessed for apoptosis by in situ end labeling of DNA fragments, cell cycle analysis of DNA content, and electron microscopy. When compared with controls, progesterone reduced and RU486 increased the percentage of apoptotic GC (p < 0.05), whereas dexamethasone had no effect. In addition, RU486 inhibited the protective effect of progesterone on GC survival (p < 0.05). Taken together, these data indicate that progesterone inhibits human GC apoptosis, and this effect is mediated through the progesterone receptor. PMID- 10818390 TI - Molecular defects causing ovarian dysfunction. AB - The functions of the hypothalamic-pituitary-ovarian and -adrenal axes are intertwined, and molecular defects in either axis may cause ovarian dysfunction. Advances in molecular genetics have allowed new insights into the pathophysiology of ovarian disorders. Specific gene mutations causing delayed puberty and/or ovarian failure, and heterosexual or isosexual precocious puberty have recently been described. The molecular insights gained into ovarian dysfunction have already led to rational therapies for some of these conditions. PMID- 10818391 TI - Follicular atresia and luteolysis. Evidence of a role for N-cadherin. AB - Studies suggest that cell-cell interactions may regulate apoptosis; and, in particular, the calcium-dependent cell adhesion molecule N-cadherin has been shown to be capable of modulating this process. Here, we review the evidence that N-cadherin is expressed by human granulosa cells (GCs) and mediates cell-cell adhesion between GCs. There is strong correlation between N-cadherin expression by granulosa or luteal cells and follicular survival in isolated follicles and archival tissue sections. There exists a strong expression of the molecule by GCs in follicles of the resting pool, of growing antral follicles, and of healthy corpora lutea. In contrast, the molecule is lost in degenerating GCs of atretic follicles and in luteal cells of the late luteal phase. Further, the experimental evidence demonstrates that cell-cell adhesion is critical to the survival of GCs and that N-cadherin-mediated cell-cell adhesion is a critical mediator of survival signals and inhibits apoptosis in these cells. Possible mechanisms by which apoptosis may be triggerred in GCs include the downregulation of N cadherin, which is mediated, at least in part, through the enzymatic cleavage of the extracellular domain of the molecule. Collectively, these observations suggest that downregulation of N-cadherin or the absence of a functional extracellular domain of the molecule prevent GC aggregation and is associated with GC apoptosis. We propose that N-cadherin-mediated GC signaling plays a central role in follicular and luteal cell survival. PMID- 10818392 TI - Molecular basis of gynecological cancer. AB - Alterations in the cellular genome affecting the expression or function of genes controlling cell growth and differentiation are considered to be the main cause of cancer. Genes that cause cancer are of two distinct types: oncogenes and onco suppressor genes. The normal proto-oncogene can be converted into an active oncogene by deletion or point mutation in its coding sequence, gene amplification, and by specific chromosome rearrangements. Mutations and abnormal expression in ras, myc, c-erbB-2, and other oncogenes have been reported in several types of gynecological cancer. Onco-suppressor genes are involved in gynecological cancer, their functions are localized in different phases of the cell cycle. Structural changes and deletions of these genes can cause cancer. Mutations in the p53, BRCA1, DCC, and PTEN genes have been reported in gynecological cancers such as ovarian, cervical, and endometrial cancer. Human papillomaviruses are of major interest because specific types (HPV-16, -18, and several others) have been identified as causative agents in at least 90% of cancers of the cervix. In this study we summarize the available information regarding the implication of specific oncogenes, onco-suppressor genes, and HPV in the development of female genital malignancies. PMID- 10818393 TI - The hypothalamic-pituitary-thyroid axis and the female reproductive system. AB - Increasing evidence derived from experimental and clinical studies suggests that the hypothalamic-pituitary-thyroid axis (HPT) and the hypothalamic-pituitary ovarian axis (HPO) are physiologically related and act together as a unified system in a number of pathological conditions. The suggestion that specific thyroid hormone receptors at the ovarian level might regulate reproductive function, as well as the suggested influence of estrogens at the higher levels of the HPT axis, seems to integrate the reciprocal relationship of these two major endocrine axes. Both hyper- and hypothyroidism may result in menstrual disturbances. In hyperthyroidism the most common manifestation is simple oligomenorrhea. Anovulatory cycles are very common. Increased bleeding may also occur, but it is rare. Hypothyroidism in girls can cause alterations in the pubertal process; this is usually a delay, but occasionally it can result in pseudo-precocious puberty. In mature women hypothyroidism usually is associated with abnormal menstrual cycles characterized mainly by polymenorrhea, especially anovulatory cycles, and an increase in fetal wastage. PMID- 10818394 TI - Thyroidopathies. AB - Pregnancy affects thyroid physiology in many ways: (a) The renal iodide clearance rate is increased, hence iodine requirements increase. (b) The fetal requirements for thyroid hormones and iodide are an additional problem. (c) Serum thyroxine binding globulin increases, thus producing an increase in the levels of total T4 and T3. (d) Chorionic gonadotropin has a thyroid-stimulating activity. This may be compensated for by a decrease in TSH, but in some cases gestational thyrotoxicosis occurs. (e) Thyroid autoimmunity usually subsides during pregnancy, but may rebound a few months after parturition, and postpartum thyroiditis may occur. Because maternal antithyroid autoantibodies cross the placenta readily, fetal and neonatal hyperthyroidism (or hypothyroidism) may develop. Pre-existing thyroid diseases are influenced. Nontoxic goiter increases in size. Iodine and/or thyroxine may be required. Graves' disease may remit. If present, antithyroid drugs should be given in small doses, and quite often they may be stopped altogether. Hypothyroid patients may require a larger T4 dose. PMID- 10818395 TI - Placental corticotropin-releasing factor. An update. AB - Corticotropin-releasing factor (CRF) produced in placenta has paracrine effects within placenta, decidua, and myometrium and endocrine effects on mother and fetus. CRF is a potent local regulator of myometrial contractility and of prostaglandin release, Recently, urocortin, a new member of the CRF family, has been localized in human placenta and membranes. Urocortin mimics some of the local effects of CRF in intrauterine tissues, that is, increase of adrenocorticotrophic hormone (ACTH) and prostagiandin release and myometrial contractility. A local CRF-BP modulates the paracrine effects of CRF and urocortin. The various CRF receptor subtypes are well distributed in placenta and membranes. CRH also acts on placental blood vasculature and has an action on fetal adrenal gland to stimulate the production of the steroid DHEA-S. In nonpregnant women, plasma CRF levels are low; they become higher during the first and second trimesters of pregnancy. A clear increase is evident at term and when CRF-BP levels decrease. Women with preterm labor show high CRF and low CRF-BP levels, supporting an involvement of this pathway in mechanism of parturition. PMID- 10818396 TI - Maternal hypothalamic-pituitary-adrenal axis in pregnancy and the postpartum period. Postpartum-related disorders. AB - During pregnancy, placenta-derived CRH increases exponentially in the plasma. Circulating levels of CRH-binding protein decrease considerably in the last trimester of pregnancy, resulting in further elevation of bioavailable plasma CRH. The adrenal glands during pregnancy gradually become hypertrophic because of the increase in ACTH, which parallels that of CRH. Thus, pregnancy is a transient period of relative hypercortisolism. The activation of the hypothalamic-pituitary adrenal axis during pregnancy has been proposed to function as a biological clock. In this model, the placenta is perceived as a stress-sensitive organ and placental CRH as a timing starter, determining a preterm, term, or postterm labor. During pregnancy, as well as during the immediate postpartum period, the hypothalamic maternal CRH secretion is suppressed, because of the circulating levels of cortisol. Hypothalamic CRH secretion normalizes within 12 weeks. This transient postpartum maternal hypothalamic CRH suppression, together with the steroid withdrawal that follows parturition, might be causally related to the mood disorders and the vulnerability to autoimmune diseases such as thyroiditis or rheumatoid arthritis often observed during the postpartum period. PMID- 10818397 TI - Clinical implications of the ovarian/endometrial renin-angiotensin-aldosterone system. AB - New organ-specific functions of angiotensin II have recently been described: the importance of its role in the regulation of secretory epithelial function in many tissues including components of the reproductive tract has been documented. The source of angiotensin II in these tissues is the reproductive tract itself, and there is considerable evidence to suggest a distinct renin-angiotensin aldosterone system in the ovary and uterus. Two main subtypes of angiotensin II receptors are recognized as angiotensin-receptor I and II, according to their sensitivity to the angiotensin II antagonists. However, the presence of angiotensin II receptors in the male and female reproductive tract suggests a multiplicity of roles that are unrelated to their primary functions or to each other. The renin-angiotensin-aldosterone system is a major determinant of sodium balance in pregnancy. More recently RT-PCR methods have revealed angiotensinogen transcription in the smooth muscle of spiral anteries of the decidua; a specific allele of this gene may be associated with hypertension in pregnancy as well as in pre-eclampsia. We investigated the evolution of plasma renin activity and aldosterone levels during normal and hypertensive pregnancy. Both were found to increase progressively during all three trimesters of normotensive pregnancy. Plasma renin activity in hypertensive women remained unchanged during all three trimesters of pregnancy. Plasma aldosterone levels in hypertensive women increased progressively during all three trimesters of pregnancy. However, plasma aldosterone levels remained significantly lower than the ones of normotensive pregnant women. These increased aldosterone levels were noticed despite unchanged renin levels. Further clinical studies investigating the renin-angiotensin aldosterone system in the pathogenesis of pregnancy hypertension are needed. A renin-independent role of aldosterone in this pathological entity is suggested. PMID- 10818398 TI - Reproductive health in female patients with beta-thalassemia major. PMID- 10818399 TI - Anemia in pregnancy. AB - Anemia is one of the most frequent complications related to pregnancy. Normal physiologic changes in pregnancy affect the hemoglobin (Hb), and there is a relative or absolute reduction in Hb concentration. The most common true anemias during pregnancy are iron deficiency anemia (approximately 75%) and folate deficiency megaloblastic anemia, which are more common in women who have inadequate diets and who are not receiving prenatal iron and folate supplements. Severe anemia may have adverse effects on the mother and the fetus. Anemia with hemoglobin levels less than 6 gr/dl is associated with poor pregnancy outcome. Prematurity, spontaneous abortions, low birth weight, and fetal deaths are complications of severe maternal anemia. Nevertheless, a mild to moderate iron deficiency does not appear to cause a significant effect on fetal hemoglobin concentration. An Hb level of 11 gr/dl in the late first trimester and also of 10 gr/dl in the second and third trimesters are suggested as lower limits for Hb concentration. In an iron-deficient state, iron supplementation must be given and follow-up is indicated to diagnose iron-unresponsive anemias. PMID- 10818400 TI - Antenatal assessment for the detection of fetal asphyxia. An evidence-based approach using indication-specific testing. AB - One of the most important advances in perinatal health care is the use of antepartum fetal testing. Antepartum fetal testing methods may include inexpensive tests such as fetal kick counts or tests that can be quite expensive such as non-stress tests, fetal biophysical profiles, and Doppler assessments as well as invasive tests such as amniocentesis or cordocentesis. Clinical experience, combined with recent literature, suggest that there is no ideal test for all high-risk fetuses and that some antepartum fetal tests may be more appropriate than others, depending on the underlying pathophysiology or the indication for testing. Because many different pathophysiological processes lead to fetal acidemia and in-utero death, indication-specific testing may be not only logical, but also cost-effective. In this article, specific guidelines of antepartum fetal testing are presented. These indication-specific guidelines are based on the underlying pathophysiological processes that place the fetus at risk and also on the need to use the fewest number of tests without compromising safety. PMID- 10818401 TI - Recent advances in respiratory care of the term neonate. AB - Persistent pulmonary hypertension is a major cause of morbidity and mortality in the term and near-term infant. Management of this condition, which is characterized by respiratory distress and cyanosis, has been greatly enhanced by inhaled nitric oxide (NO) therapy. The following treatment regime is suggested: Conventional ventilation should be used initially, and hyperventilation should be avoided. Surfactant should be administered early, preferably within 6 hours of diagnosis. If conventional ventilation fails, the next step is high-frequency ventilation or inhaled NO. Some infants who do not respond to inhaled NO when administered by conventional ventilation will respond to NO delivery via a high frequency ventilator. If all of these therapies fail, extracorporeal membrane oxygenation (ECMO) should be considered. By the use of this approach, the mortality from PPH has been considerably reduced, and concerns today relate primarily to morbidity, particularly long-term neurologic outcome and chronic lung disease resulting from ventilation and barotrauma. PMID- 10818402 TI - Mechanisms of perinatal cerebral injury in fetus and newborn. AB - Cerebral hypoxia in the fetus and newborn results in neonatal morbidity and mortality as well as long-term sequelae such as mental retardation, seizure disorders, and cerebral palsy. In the developing brain, determinants of susceptibility to hypoxia should include the lipid composition of the brain cell membrane, the rate of lipid peroxidation, the presence of antioxidant defenses, and the development and modulation of excitatory amino acid neurotransmitter receptors such as the N-methyl-D-aspartate (NMDA) receptor, the intracellular Ca2+, and the intranuclear Ca(2+)-dependent mechanisms. In addition to the developmental status of these cellular components, the response of these potential mechanisms to hypoxia determines the fate of the hypoxic brain cell in the developing brain. Using electron spin resonance spectroscopy of alpha-phenyl N-tert-butyl-nitrone spin adducts, studies from our laboratory demonstrated that tissue hypoxia results in increased free radical generation in the cortex of fetal guinea pigs and newborn piglets. Pretreatment with MgSO4 significantly decreased the hypoxia-induced increase in free radical generation in the term fetal brain. We also showed that brain tissue hypoxia modifies the NMDA receptor ion-channel recognition and modulatory sites. Furthermore, a higher increase in NMDA receptor agonist-dependent Ca2+ in synaptosomes was demonstrated. The increase in intracellular Ca2+ may activate several enzymatic pathways such as phospholipase A2 and metabolism of archidonic acid by cyclooxygenase and lipoxygenase, conversion of xanthine dehydrogenase to xanthine oxidase by proteases, and activation of nitric oxide synthase. Using inhibitors of each of these enzymes such as cyclooxygenase (indomethacin), lipoxygenase (nordihydroguaiaretic acid), xanthine oxidase (allopurinol), and nitric oxide synthase (N-nitro-L-arginine), studies have shown that these enzyme reactions result in oxygen free radical generation, membrane peroxidation, and cell membrane dysfunction in the hypoxic brain. Specifically, generation of nitric oxide free radicals during hypoxia may lead to nitration and nitrosylation of specific membrane proteins and receptors, resulting in dysfunction of receptors and enzymes. We conclude that hypoxia-induced modification of the NMDA receptor leading to increased intracellular Ca2+ results in free radical generation and cell injury. We suggest that during hypoxia the increased intracellular Ca2+ may lead to increased intranuclear Ca2+ concentration and alter nuclear events including transcription of specific apoptotic genes and activation of endonucleases, resulting in programmed cell death. PMID- 10818403 TI - Angiogenic factors in the perinatal period: diversity in biological functions reflected in their serum concentrations soon after birth. AB - These studies investigated whether serum levels of the angiogenic factors angiogenin, basic fibroblast growth factor (bFGF), and vascular endothelial growth factor (VEGF) change soon after birth due to the elimination of the placenta and to diminished angiogenic but increased adaptational demands in extrauterine life. Also investigated was whether serum levels correlate with sex, birth weight, or mode of delivery. Serum from healthy mothers and their healthy full-term infants at birth (umbilical cord, UC), day 1 (N1) and day 4 (N4) postpartum was analyzed by enzyme immunoassays. Angiogenin levels were higher in maternal serum and rose significantly from UC to N1 and N4, possibly because of the elimination of the placenta, which produces an angiogenin inhibitor. bFGF and VEGF maternal levels were lower than fetal and neonatal ones. Although neonatal bFGF levels did not differ from fetal levels, possibly reflecting diminished angiogenesis ex utero, VEGF levels increased in neonatal serum, possibly signifying adaptational demands. Neither factor depended on sex, mode of delivery, or birth weight. Thus, significant differences from normal reference values of the studied factors might reflect ill-defined situations of the placenta and fetus/newborn serving as early diagnostic markers. PMID- 10818404 TI - Role of leptin in reproduction. AB - Leptin is a 16-kDa adipocyte-secreted protein the serum levels of which reflect mainly the amount of energy stores but are also influenced by short-term energy imbalance as well as several cytokines and hormones. Leptin, by binding to specific receptors, alters the expression of several hypothalamic neuropeptides that regulate neuroendocrine function as well as energy intake and expenditure. More specifically, accumulating evidence suggests that this hormone may serve to signal to the brain information on the critical amount of fat stores that are necessary for LHRH secretion and activation of the hypothalamic-pituitary-gonadal axis. Rising leptin levels have been associated with initiation of puberty in animals and humans and normal leptin levels are needed for maintenance of menstrual cycles and normal reproductive function. Moreover, circadian and ultradian variations of leptin levels are associated with minute to minute variations of LH and estradiol in normal women. Falling leptin levels in response to starvation result in decreased estradiol levels and amenorrhea in subjects with anorexia nervosa or strenuously exercising athletes. In addition, leptin has a potentially important role during pregnancy and in the physiology of the neonate. Finally, recent evidence suggests that leptin may influence ovarian steroidogenesis directly, but the exact role of intraovarian leptin action in the physiology and pathophysiology of the human reproductive system needs to be further elucidated. PMID- 10818405 TI - The potential role of intraovarian factors on ovarian androgen production. AB - The circuit of gonadotropins (FSH, LH) and ovaries (theca and granulosa cells) in ovarian estrogen and androgen production is well established. Recent research has revealed an intraovarian network that may ultimately prove relevant to the understanding of ovarian hyperandrogenism. Most of these substances, such as growth factors and cytokines, do not have independent effects on basal androgen production, but exhibit their regulatory potential by modulating hCG- or LH stimulated steroid production. Precise understanding of the regulatory role of intraovarian factors in ovarian androgen production would shed new light on the pathophysiology and therapy of hyperandrogenemic excess in women. PMID- 10818406 TI - The genetics of obesity. Lessons for polycystic ovary syndrome. AB - Both polycystic ovary syndrome (PCOS) and obesity are common disorders with a complex phenotype. Both are presumably heterogeneous in etiology. Understanding the genetics of obesity, which has a longer and richer history, may therefore illuminate the genetics of PCOS, where major projects are now underway. Obesity may be the penultimate condition in which the effects of heredity and environment will forever mingle. Most obesity mutations identified to date (with the exception of the Agouti mutation) are inherited in an autosomal recessive manner. Therefore, it is unlikely that such mutations, even when identified in a human population, could explain only a fraction of the cases that make up the high prevalence of both of these disorders. Although the mouse models of single gene defects causing obesity contain many similar aspects of the PCOS phenotype such as obesity and subfecundity, there is no mouse model that mimics all aspects of the syndrome, especially the circulating androgen excess. This elevation in circulating androgens may be the sine qua non of the syndrome as indicated by our findings in sisters of PCOS probands that hyperandrogenemia may be the distintinctive reproductive phenotype. Isolation of PCOS and obesity genes may allow the development of targeted interventions that will lead to effective and safe treatment of both obesity and PCOS. PMID- 10818407 TI - Insulin sensitizers and antiandrogens in the treatment of polycystic ovary syndrome. AB - The heterogeneous origin of polycystic ovary syndrome (PCOS) has been demonstrated by several studies. Abnormalities in steroidogenesis and metabolism are present, but the exact link between these two pathologic features remains to be clarified. In clinical practice, more than one therapeutic approach for the treatment of this syndrome has been proposed over the last few decades. Because hyperandrogenism and hyperinsulinemia contribute to a different degree to the phenotype of PCOS, therapeutic efforts have focused on agents that could treat or modify the clinical manifestations of these disorders. Antiandrogens as a sole treatment or combined with oral contraceptives are considered the treatment of choice for the manifestations of hyperandrogenemia, but there is no agreement about their efficacy on the metabolic sequelae of PCOS (insulin resistance, hyperinsulinemia, dislipidemia). Furthermore, the improvement of insulin sensitivity by insulin sensitizers may be of therapeutic value directly and/or indirectly in the management of clinical manifestations of hyperinsulinemia and hyperandrogenemia. PMID- 10818408 TI - Optimizing estrogen replacement in adolescents with Turner syndrome. PMID- 10818409 TI - Contraception in women at high risk or with established cardiovascular disease. AB - Oral contraceptives are one of the most effective and widely used reversible contraceptive methods. Over 90 million women worldwide, including over 44 million in developing countries, are now using oral contraceptives. Despite their advantages, there is concern about the links between combined oral contraceptives and the risk of cardiovascular disease. The risk attributable to oral contraceptive use in women < 35 years of age is small, even if they smoke, but there are substantially increased risks in older women who both smoke and use oral contraceptives. Differences between oral contraceptive types in the relative risk of venous thromboembolism contribute little to the total cardiovascular mortality associated with oral contraceptive use, even though the total number of cardiovascular events is increased. It is important to consider the user's age and smoking status when determining oral contraceptive-attributable risks. Hormonal oral contraceptives have changed and now contain lower doses of estrogen and progestagen. PMID- 10818410 TI - Oral contraceptives' effects on the vascular component. Thrombophilic parameters. PMID- 10818411 TI - Dysmenorrhea. AB - Dysmenorrhea presents as painful periods that start two to three years after menarche. The pain usually begins when the bleeding starts and lasts for 48-32 hours. The cause of menstrual cramps and associated symptoms in primary dysmenorrhea is related to prostaglandin production. In secondary dysmenorrhea, there is documented pelvic pathology that causes the painful menstrual cramps, and treatment is cause related. Available treatments for primary dysmenorrhea- NSAIDS, oral combined contraceptives, beta-blockers, psychotherapeutic methods, and cervical dilatation--are discussed. PMID- 10818412 TI - Combined oral contraceptive treatment of adolescent girls with polycystic ovary syndrome. Lipid profile. AB - The clinical signs and neuroendocrine features of adolescents with polycystic ovary syndrome (PCOS) resemble those found in adult women with PCOS. These adolescent patients are candidates for long-term treatment by one of the different therapeutic approaches that have been proposed. It is therefore essential that the treatment does not induce unfavorable metabolic effects. We investigated and compared the effects of cyproterone acetate (CA) and desogestrel (D), as part of combined oral contraceptives (COC), on lipid metabolism and hirsutism in adolescents with PCOS. Twenty-four girls with clinical signs of PCOS were recruited. They were all hyperandrogenic and euthyroid and had normal prolactin plasma levels. Non-classic congenital adrenal hyperplasia was ruled out by the ACTH stimulation test. Blood samples were obtained for sex hormone binding globulin (SHBG), total cholesterol (TC), LDL cholesterol, HDL cholesterol, triglycerides (TGs), apolipoproteins A-I, A-II, B, and lipoprotein (a) (Lp(a)) measurements. After the initial examination, therapy was initiated in a randomly selected order (12 and 12 patients were treated daily by 2 mg CA and 0.150 mg D, respectively, plus 0.035 and 0.030 mg ethinylestradiol, respectively). The degree of hirsutism and the lipid profile were reevaluated every 3 months after initiation of therapy for 1 year. Our data show that after 12 months of treatment with the D or CA COC, the Ferriman-Gallway hirsutism score decreased and TC and LDL-C increased. TGs and HDL-C were raised significantly in the CA COC group, whereas apolipoprotein A1 increased during D COC treatment. The atheromatic indices did not change. These data suggest that treatment of adolescent girls with PCOS is comparably effective with the two contraceptive formulations and that the desogestrel COC could be considered in the treatment of adolescent PCOS patients because it does not have side effects on lipid metabolism. PMID- 10818413 TI - Laparoscopic approach to ovarian cysts in women over 40 years of age. PMID- 10818414 TI - Management of hydrosalpinx: reconstructive surgery or IVF? AB - Tubal disease remains the most important factor in female infertility. Many investigators reported that patients with hydrosalpinx had a decreased clinical pregnancy rate and an increased miscarriage rate, resulting in a decreased ongoing pregnancy rate when compared to that of patients with other types of tubal disease. Different studies showed a deleterious effect of the presence of hydrosalpinx on the outcome of in vitro fertilization-embryo transfer, because toxic agents flowing from the hydrosalpinx to the uterus impair the implantation rate. Operative laparoscopy is effective in the treatment of hydrosalpinges (Stage I or II). Fertility outcome is related to tubal damage. Patients with Stage III and IV disease should be managed from the beginning with in vitro fertilization. Excision of hydrosalpinx(-ges) improves the pregnancy potential after in vitro fertilization. We believe that assisted reproductive technology and reproductive surgery can be complementary. The development of laparoscopic surgery and in vitro fertilization improved the pregnancy rate in patients with tubal factor infertility. PMID- 10818415 TI - Adhesions: laparoscopic surgery versus laparotomy. AB - This study was undertaken to assess the effectiveness in pregnancy rates of microsurgery and operative laparoscopy in adhesiolysis. Adhesions were found to be the sole infertility factor in 15% of our patients. One hundred and ninety infertile patients with periadnexal adhesions as the only cause of their infertility were treated by microsurgery (86) or operative laparoscopy (104) and were followed up for 24 months. Our results indicate that advanced laparoscopic surgery in general is as effective as microsurgery in healthy infertile patients with adhesions but offers some advantages in comparison to laparotomy. Factors that adversely affect the postoperative success rates are the age of the women, the duration of infertility, and the severity of the adhesions. PMID- 10818416 TI - Chlamydia screening--yes, but of whom, when, by whom, and with what? AB - The importance of screening programs in reducing the prevalence of genital chlamydial infections is stressed by the fact that the majority of infected persons are more or less asymptomatic. The use of oral contraceptives may mask infections affecting the upper genital tract. This imposes selective screening and rescreening of women with a history of pelvic inflammatory disease. The recent knowledge that vaginal introital samples will provide a detection rate equal to or even higher than that of cervical samples collected in the same women opens up the possibility of screening women in health units lacking a gynecological examination chair. It also opens up the possibility of outpatient screening programs, for example, home sampling and mailing samples to laboratories that will perform analyses. The use of nucleic acid-based assays means increased sensitivity and specificity compared with earlier used techniques such as ELISA. These former methods can also be used in low-prevalence populations with acceptable positive predictable value, but may be misleading if used in post-therapy check-ups because the antigen may persist in microbiologically cured cases. PMID- 10818417 TI - Update on Chlamydia trachomatis. AB - Chlamydia trachomatis is one the most important sexually transmitted diseases; it can cause serious sequelae despite the absence of symptoms in some people. It's estimated that about 25% of women who have acute salpingitis become infertile, and chlamydial infection is the commonest cause. The introduction of screening programs for its detection are still a topic of discussion. The literature shows that the total cost of examination and treatment of complications known to be associated with genital chlamydial infection (PID, chronic pelvic pain, tubal factor infertility) is generally higher than the total cost of a large-scale Chlamydia screening program. The selection of a diagnostic test for detection of chlamydial genital infection depends on availability, local expertise, and prevalence of Chlamydia trachomatis in the test population. Cell culture is too expensive in nonendemic regions, so the use of non-culture techniques is very attractive. PCR (polymerase chain reaction) and LCR (ligase chain reaction) are actually the two most commonly used alternatives to conventional methods for detecting STD agents. In fact, PCR and LCR have proved useful for detection of Chlamydia trachomatis in cervical and urethral samples both in symptomatic and asymptomatic women. Recently, testing of first-void urine (FVU) specimens with these techniques has shown that the amplification tests are as sensitive as tests with endocervical swab cultures. PMID- 10818418 TI - PID: clinical and laparoscopic aspects. AB - Clinical signs of pelvic inflammatory disease (PID) are not constant and are often limited to slight pelvic pain. Laparoscopy can lead to a rapid and correct diagnosis of PID. Intrapelvic bacteriologic samples can be obtained so as to administer the proper antibiotic. The exact nature of the lesions can be evaluated, and in severe cases, recent abscesses can be treated with good results for fecundity. Because the results in cases of long-standing abscess are not so good, laparoscopy should be performed at the onset of infection and not be reserved until after some weeks of inefficient medical treatment, especially in young women who have not completed their family. In primary chronic salpingitis, the lack of any clinical signs usually leads to a delay in diagnosis until women consult for fertility problems. The ideal point would be to detect some biologic or clinical change that may lead to diagnosis such as a positive anti-Chlamydia trachomatis (CT) serology or, in the future, positive anti-CT Hsp 60 antibody could be the key to detecting and treating silent salpingitis in young women, CT being the main microorganism involved in chronic salpingitis. Screening for C. trachomatis low genital tract infection is mandatory in young people in order to control the epidemic. PMID- 10818419 TI - Conservative management of PID. AB - The goals in the management of pelvic inflammatory disease (PID) are not only treatment of the infection and prevention of immediate complications, but also prevention of its long-term consequences. There are criteria for hospitalization, but patients who do not meet them can be safely treated as outpatients. A variety of sexually transmitted and other microorganisms can cause this infection, but the most important are Chlamydia trachomatis and Neisseria gonorrhoeae. Regimens with activity against gonococci, chlamydiae, streptococci, gram-negative bacteria, and anaerobes should be administered. Several such antimicrobial regimens have shown very good clinical and microbiologic efficacy. However, their efficacy in preventing long-term complications, such as infertility, has not been established. Close follow-up is an important part of management. Evaluation of male sexual partners is imperative to prevent reinfection. Better diagnostic techniques and treatment modalities for PID must be developed to prevent its long term consequences. PMID- 10818420 TI - Surface morphology of the human endometrium. Basic and clinical aspects. AB - The human endometrium is an extremely sensitive target for steroid hormones. During the menstrual cycle, this tissue undergoes dynamic changes that are reflected on the surface morphology of the epithelium and that can be followed by scanning electron microscopy. The morphologic changes peak at the midsecretory phase, with the formation of the so-called pinopodes. Increasing evidence suggests that these pinopodes are accurate markers for endometrial receptivity, and their detection may be of high clinical utility in the preparation of endometrium before embryo transfer. This article recapitulates published figures of endometrial ultrastructure and presents some unpublished data from ongoing studies. PMID- 10818421 TI - Biological factors in culture media affecting in vitro fertilization, preimplantation embryo development, and implantation. AB - Optimal culture conditions are of paramount importance for in vitro fertilization of gametes, preimplantation embryo development, and implantation for all species. Water is the basis of all culture media, and ultrapure water should be employed. The main energy sources of a medium are lactate, pyruvate, and glucose. The concentrations of the first two vary in different media, whereas the latter is necessary mainly for the later stages (morula to blastocyst) of development. A fixed nitrogen source is essential for implantation embryo development whether this is provided by amino acids, albumin, or serum. Suboptimal culture conditions can block development. Pronuclear zygotes of most species (but not human) arrest at some point between the two-cell and the 16-cell stage. Modifying culture conditions can lead the embryos to develop through this block. Hypoxanthine also causes a two-cell block to mouse pronuclear zygotes, and this again depends largely on culture conditions. Simple culture media are bicarbonate-buffered systems with pyruvate, lactate, and glucose. Complex media, such as Ham's F-10, contain in addition amino acids and other elements found in serum. Human tubal fluid simulates the fallopian tube microenvironment. EDTA, gonadotropins, growth factors, and other substances can be included in the media to stimulate development. Coculture of embryos with oviductal cells has shown promising results. PMID- 10818422 TI - Intracytoplasmic sperm injection. Survey of world results. AB - The widespread application of intracytoplasmic sperm injection (ICSI) has raised concern about the efficacy and safety of this novel technique. The European Society of Human Reproduction and Embryology (ESHRE) has established an ICSI Task Force to collect annually the clinical results, the outcome of pregnancy, and the follow-up of children after ICSI using ejaculated, epididymal, and testicular sperm in order to address these important issues in a relatively short time. Over a 3-year span (1993-1995), the number of centers for ICSI increased from 35 to 101, and the total number of ICSI cycles per year rose from 3,157 to 23,932. The incidence of oocytes damaged by the procedure remained low (< 10%), whereas the fertilization rates obtained with ejaculated, epididymal, and testicular spermatozoa for 1995 were 64%, 62%, and 52%, respectively. Thus, 86-90% of the couples had embryo transfer, and the viable pregnancy rate was 21% for ejaculated, 22% for epididymal, and 19% for testicular sperm, while the incidence of multiple gestations was 29%, 30%, and 38%, respectively. It is noteworthy that no difference was found in ICSI results concerning the etiology of azoospermia, for example, obstructive (congenital or acquired) or nonobstructive. Furthermore, 3,149 transfers of frozen-thawed embryos after ICSI with ejaculated, epididymal, or testicular sperm were performed, and in 11%, 9%, and 7% of them, respectively, a viable pregnancy was achieved. The ICSI results were similar during this 3-year period, irrespective of the origin of the sperm. The perinatal outcome of children born after ICSI was not different from that after in vitro fertilization or natural conception and was only affected by multiplicity. Moreover, the incidence of major or minor malformations was not increased, but the chromosomal, especially the sex chromosomal, aberration rate was slightly elevated (approximately 2%). Therefore, ICSI has opened new horizons in the treatment of male infertility. The achievement of pregnancy after ICSI using ejaculated, epididymal, or testicular sperm is very satisfactory. The procedure seems to be safe, but further follow-up of the children is necessary to more accurately assess its safety. PMID- 10818423 TI - Assessing the outcome of IVF. PMID- 10818424 TI - Autoimmune antiovarian antibodies and their impact on the success of an IVF/ET program. AB - In previous papers, we referred to studies of the influence of antiovarian autoantibodies on menstrual cycle disorders in adolescent girls. We examined autoantibodies against ooplasma, zona pellucida, membrana granulosa, theca folliculi interna, and lutein cells. In infertile women in the IVF/ET program, we studied the positivity of antiovarian antibodies and cytokines, namely, TNF-alpha and IL-1 beta, in follicular fluid correlated with the following subgroups, characterized by the outcome of in vitro fertilization, as follows: G, pregnant; F, fertilized; N, nonfertilized; and O, no oocyte gained. The presence of autoantibodies corresponds to the success or failure of the IVF/ET program. Our results support the hypothesis that antiovarian autoantibodies play an important role in both the endocrine and the reproductive function of the human ovary and that it can influence them negatively. PMID- 10818425 TI - Ethical aspects of reproduction in the next century. AB - Doubtless, the technological advancements achieved in the current century in the area of young women's reproductive health care have also led to the evolution of very important ethical issues that will have to be dealt with in the coming century. Abortion, perhaps the most controversial issue of all, continues to raise a number of ethical issues related to the rights of the women versus the rights of the fetus, which, in addition to the risk of sexually transmitted diseases including the human immunodeficiency virus, emphasize the need for adequate family planning and sexual education. Genetic testing for late-onset diseases, disease susceptibilities, and carrier status may offer medical or psychological benefits; however, several complex ethical, legal, and social issues have been revealed with the advent of this new information. New family structures deviating from the traditional heterosexual couple, consisting of either single or lesbian mothers, have appeared, raising serious disputes regarding the welfare of the child. Important demographic changes are expected in the world population in the 21st century, characterized mainly by a significant increase of the older age groups. PMID- 10818426 TI - The perimenopausal transition. PMID- 10818427 TI - Premature ovarian failure is not premature menopause. AB - Normal menopause occurs at an average age of 50 and results from ovarian follicle depletion. Normal menopause is an irreversible condition, whereas premature ovarian failure is characterized by intermittent ovarian function in half of these young women. These young women produce estrogen intermittently and sometimes even ovulate despite the presence of high gonadotropin levels. Indeed, pregnancy has occurred after a diagnosis of premature ovarian failure. On pelvic ultrasound examination, follicles were equally likely to be detected in patients more than 6 years after a diagnosis of premature ovarian failure as in patients less than 6 years after the diagnosis. Thus, the probability of detecting a follicle appears to remain stable during the normal reproductive lifespan of these young women. Indeed, pregnancy was reported in a 44-year-old woman 16 years after a diagnosis of premature ovarian failure. No treatment to restore fertility in patients with premature ovarian failure has proved to be safe and effective in prospective controlled studies. Theoretically, these unproved therapies might even prevent one of these spontaneous pregnancies from occurring. PMID- 10818428 TI - Osteopenic syndromes in the adolescent female. AB - Low bone density in growing girls and mature young women is usually a finding that needs an explanation and further clinical investigation. Population-based epidemiologic studies on osteoporosis in young persons do not exist. As a disease, osteoporosis among children and adolescents is rate, and since 1965 only 100 cases of idiopathic juvenile osteoprososis have been reported. When osteoporosis occurs in children, it is usually secondary to an underlying medical disorder (e.g., anorexia nervosa, leukemia) or to medications, but occasionally no identifiable primary cause can be detected. It may also be the result of a genetic disorder such as osteogenesis imperfecta. On the other hand, osteopenia in growing and young persons seems much commoner and needs further investigation. Adolescence is a period of increased calcium requirement, and girls with an underlying bone disease are at higher risk for bone demineralization. An additional point of interest is the changes in the geometry of bones through their continuous adaptation to simultaneous skeletal and muscular growth. Bones, through the mechanostat mechanism, adapt to mechanical loading by differentiating their geometry. A recent finding in this direction is that before and during the teenage years there is an environmental effect of physical activity and nutrition on hip geometry. Another important finding is an age-dependent increase in bone cross-sectional area and bone strength index in the absence of an increase in volumetric spongiosa bone density and cortical bone density. Girls, in comparison to boys, deposit more calcium in their bones during puberty, thus probably preparing their skeleton for the forthcoming events of pregnancy and lactation. PMID- 10818429 TI - Idiopathic juvenile osteoporosis. AB - Osteoporosis in childhood is uncommon, and it may be secondary to a spectrum of diverse conditions. When such causes have been excluded, some patients remain who have a congenital disease (osteoporosis imperfecta) or a disease in which the etiology is obscure, called idiopathic juvenile osteoporosis (IJO). The cause of IJO is unknown, and the diagnosis is based both on the exclusion of other diseases and on its own positive fractures. The basic strategy of treatment is to protect the spine until remission occurs. Sex steroids are contraindicated. Bisphosphonates, calcitriol, fluoride, and calcitonin have been administered therapeuticlly, but the results were equivocal. Usually the disease remits by itself. PMID- 10818430 TI - Alternatives to estrogen replacement therapy. PMID- 10818431 TI - Estrogen replacement therapy in the management of osteopenia related to eating disorders. AB - The effect of hormone replacement therapy on the bone mineral content of hypoestrogenic subjects depends on the pathogenesis of the disease as well as on the dosage and route of administration. This is particularly true in hypoestrogenism related to eating disorders. We present a longitudinal study of 26 young women with diet-induced amenorrhea compared with a group of subjects with POF. The study protocol included the quantification of weight loss, the endocrine profile (follicle-stimulating hormone, luteinizing hormone, prolactin, E2, FT3, FT4, thyroid-stimulating hormone, and cortisol), the evaluation of markers of bone turnover (GLA, OSTK-PR, ALP, OHP, and DPYR), and spinal bone density by DEXA at observation and after weight recovery. No hormone replacement therapy was administered. Mean BMD and Z scores before and after recovery do not differ significantly; OHP and DPYR appear significantly higher during basal evaluation, whereas GLA and ALP do not. Data on the impact of oral contraceptive use on bone mineral density are controversial. We particularly discuss the question of long-term treatment with 20 micrograms ethinyl estradiol pills on peak bone mass acquisition during adolescence. PMID- 10818432 TI - Hormone replacement therapy and breast cancer risk. AB - The female breast is subject to a lifetime of hormonal control. After menopause, breast tissue becomes quiescent when estrogens drop to low levels. Menopause associated hormonal decreases produce short- and long-term consequences that can be treated successfully by hormone replacement therapy (HRT). Despite the beneficial effects of HRT, the potential risk of breast cancer is a concern of both women and physicians. The available data indicate that HRT administered for longer than 5 years moderately increases the risk of breast cancer, but overall the benefits outweight the potential risk. PMID- 10818433 TI - Gonadotropin-releasing hormone analogues in the management of precocious puberty. PMID- 10818434 TI - Current and potential application of GnRH agonists in gynecologic practice. AB - The development of GnRH-a (analogues or agonists) is a major leap forward in the treatment of various hormone-dependent diseases in medicine. Their introduction in reproductive endocrinology, in in vitro fertilization/embryo transfer (IVF/ET) and other assisted reproduction techniques had a revolutionary impact. They have been effective in other gynecologic conditions including fibroids, endometriosis, anovulatory disorders, precocious puberty, dysfunctional uterine bleeding, and operative hysteroscopy. Medical castration induced by GnRH-a has become first line therapy in metastatic breast cancer. Their long-term use, though, has been associated with a variety of adverse effects such as bone loss and decreased cardioprotection. "Steroid add-back" therapy in these cases apparently is an effective alternative. It may allow their safe long-term application beyond 6 months, averting unpleasant side-effects and maintaining bone mass and cardioprotection. PMID- 10818435 TI - Preface: the omnific lysophospholipid growth factors. PMID- 10818436 TI - Development of our current understanding of bioactive lysophospholipids. AB - Lysophosphatidic acid (LPA) serves as the prototypic lysophospholipid mediator that acts through G-protein-coupled receptors to evoke a host of responses in numerous target cells. The hormone- and growth-factor-like activities of LPA, mediated by distinct G proteins, were discovered about 10 years ago. Since then, considerable progress has been made in our understanding of LPA receptor signaling, culminating in the recent identification of a growing family of heptahelical receptors specific for LPA and the structurally related lysolipid, sphingosine-1-phosphate (S1P). In addition to stimulating Gi-Ras-mediated cell proliferation, LPA and S1P induce rapid G alpha 12/13-RhoA-mediated cytoskeletal changes underlying such diverse responses as neurite retraction, cell rounding, and enhanced tumor cell invasiveness. LPA also triggers inhibition of gap junctional communication. This overview focuses on how our understanding of LPA as an intercellular lipid mediator has developed during the last decade. PMID- 10818437 TI - Group IV cytosolic phospholipase A2 activation by diacylglycerol pyrophosphate in murine P388D1 macrophages. AB - Diacylglycerol pyrophosphate (DGPP) is a novel phospholipid identified in yeast, bacteria, and plants, but not yet in mammalian cells. Given its structural resemblance to other phospholipid-activating molecules, such as lysophosphatidate, phosphatidate, and diacylglycerol, it was questioned whether DGPP was capable of activating macrophages to release arachidonic acid (AA) metabolites such as the prostaglandins. It has been found that DGPP is able to potently stimulate prostaglandin production in the murine cell line P388D1 by a mechanism that involves activation of the cytosolic Group IV phospholipase A2 (cPLA2). Our results demonstrate that DGPP possesses macrophage-activating-factor properties and suggest a role for this novel compound in the inflammatory response. PMID- 10818438 TI - Sphingosine-1-phosphate signaling via the EDG-1 family of G-protein-coupled receptors. AB - The bioactive lipid sphingosine-1-phosphate (SPP) is abundantly formed and released during the activation of platelets by thrombotic stimuli. Once exported, SPP interacts with the G-protein-coupled receptors (GPCR) of the EDG-1 family. SPP binds to EDG-1 with the dissociation constant of approximately 8 nM and induces signal transduction events such as mitogen-activated protein kinase (MAP kinase) activation, decrease of cAMP levels, remodeling of the actin cytoskeleton, among others. EDG-1 is a prototypical member of a large family of GPCRs that interact with glycero- and sphingolysolipid phosphates, namely, SPP and lysophosphatidic acid (LPA). Three other GPCRs, trivially termed EDG-3, EDG 5, and EDG-8, are also high-affinity receptors for SPP. The four SPP receptor subtypes regulate different intracellular signal transduction pathways. In vascular endothelial cells, cooperative signaling between EDG-1 and EDG-3 subtypes of SPP receptors results in adherens junction assembly, cell survival, morphogenesis into capillary-like networks, and angiogenesis. SPP acts distinctly, albeit cooperatively, with polypeptide angiogenic factors, resulting in the formation of mature neovessels. Thus SPP signaling as an extracellular mediator via the EDG-1 family of GPCRs may be a heretofore unrecognized mechanism for the regulation of angiogenesis and vascular endothelial cell function. PMID- 10818439 TI - Molecular identification and characterization of G protein-coupled receptors for lysophosphatidic acid and sphingosine 1-phosphate. AB - Lysophospholipid mediators, lysophosphatidic acid (LPA) and sphingosine 1 phosphate (S1P), play important roles in diverse biological processes, including cell proliferation, survival, cytoskeleton changes, migration, wound healing, angiogenesis, tumor invasion, and embryonic development. The recent breakthrough in cloning and identification of several G-protein-coupled receptors for LPA and S1P has provided tools to dissect the complex mechanisms by which these lysophospholipids exert their (patho)physiological functions. Here, strategies and efforts in cloning and identifying the Edg family receptors for LPA and S1P are reviewed. Reporter gene assays used to take advantage of signaling properties of LPA and S1P, and to overcome intrinsic difficulties unique to this system, are described. From these experimental approaches, important lessons can be learned and applied to future studies of signal transduction of lysophospholipid receptors. PMID- 10818440 TI - Pharmacological characterization of phospholipid growth-factor receptors. AB - The phospholipid growth-factor (PLGE) terminology is proposed to describe a group of endogenous glycerol- and sphingolipid mediators that regulate cell proliferation through plasma membrane receptors. In addition to LPA and SPP, multiple PLGFs are present in blood plasma and serum. PLGF activity is regulated by its stimulus-coupled production and by endogenous inhibitors. In addition to LPA and SPP, alkenyl-glycerophosphate, cyclic-phosphatidic acid, and sphingosylphosphorylcholine were detected in biological fluids using mass spectrometry. Heterologous desensitization studies indicate the expression of multiple LPA-activated receptors in a variety of cell types, which are differentially activated by the different PLGFs. Northern blot and RT-PCR results reinforce the coexpression of PSP24 alpha and different members of the EDG1-7 receptors in the same cell. Stable heterologous expression of the PSP24 alpha, EDG2, and EDG4 receptors in HEK293 cells show distinct PLGF specificities and dose-response properties for each receptor subtype. Thus, both the controlled availability of the different agonists/inhibitors and the regulated expression of their receptors regulate the biological effects of PLGFs. PMID- 10818441 TI - Sphingosine 1-phosphate: a ligand for the EDG-1 family of G-protein-coupled receptors. AB - Ample evidence indicates that sphingosine-1-phosphate (SPP) can serve as an intracellular second messenger regulating calcium mobilization, cell growth, and survival. Moreover, the dynamic balance between levels of the sphingolipids metabolites, ceramide, and SPP, and consequent regulation of opposing signaling pathways, is an important factor that determines whether a cell survives or dies. This ceramide/SPP rheostat is an evolutionarily conserved stress regulatory mechanism influencing growth and survival of yeast. In addition, SPP also has been identified as the ligand for the G-protein-coupled receptors EDG-1, -3, -5, and -6. Binding of SPP to EDG-1 regulates chemotaxis and in vitro angiogenesis of endothelial cells, whereas EDG-5, and possibly EDG-3, are likely the cell surface receptors responsible for cell rounding and neurite retractions induced by SPP. Hence, the studies identify a family of highly specific SPP receptors that are capable of mediating different biological responses. Thus, it is suggested that SPP is a prototype for a novel class of lipid mediators that act both extracellularly as ligands for cell surface receptors and intracellularly as second messengers. Recently, sphingosine kinase was purified to homogeneity and the first mammalian sphingosine kinase, the enzyme responsible for the formation of SPP, was cloned. The studies should provide the necessary tools to develop insight into the biological roles of this important bioactive sphingolipid. PMID- 10818442 TI - Phospholipase D. AB - Phospholipase D is an ubiquitous enzyme that hydrolyzes phosphatidylcholine to phosphatidic acid and choline. Its cellular actions are related to the production of phosphatidic acid and include alterations to cell growth, shape, and secretion. There are two mammalian phospholipase D genes whose products (PLD1 and PLD2) are alternatively spliced. Both forms have two highly conserved HKD motifs that are essential for catalysis and dimerization. PLD1 is regulated in vitro and in vivo by protein kinase C and small GTPases of the Rho and ARF families, whereas PLD2 shows a higher basal activity with little or no response to these proteins. The cellular locations and specific functions of the two PLD isoforms remain to be established. PMID- 10818443 TI - A novel polyisoprenyl phosphate signaling cascade in human neutrophils. AB - Activation of neutrophil (PMN) surface receptors can evoke inflammation and tissue injury via aberrant release of excess effectors. The molecular mechanisms involved in host protection and control of PMN responses have yet to be defined. As Billah and coworkers (1989), and Exton (1997), for example, have pointed out, phospholipase D (PLD) signaling is known to play a pivotal role in PMN activation. Here, we determined the relationship between polyisoprenyl phosphate (PIPP) remodeling and PLD signaling and their impact in activation of PMN receptors by "pro-inflammatory" (leukotriene B4), and "anti-inflammatory" (aspirin-triggered lipoxinA4) ligands. Activation of the leukotriene B4 receptor initiated a rapid (within seconds) decrement in presqualene disphosphate (PSDP), activation of PLD and production of superoxide anions. This contrasts with activation of the LXA4 receptor by an aspirin-triggered lipoxin A4 mimetic that before leukotriene B4 gave an inverse relationship with rapidly increasing PSDP levels, and inhibition of both PLD activity and superoxide generation. PSDP proved to be a potent and direct-acting inhibitor of PLD (rhPLD1b:Ki = 5.9 nM), a property not shared by structurally related endogenous lipids. This PIPP also interacted with Src homology domains, selectively targeting SH2 and not SH3 domains. These results indicate a role for ligand-driven rapid PIPP remodeling as an early switch and "stop" signaling event that controls PMN. Moreover, they indicate that PSDP directly down-regulates PMN signaling events via select protein-target interactions controlling intracellular responses relevant in inflammation. PMID- 10818444 TI - Lipid phosphate phosphatase-1 in the regulation of lysophosphatidate signaling. AB - Mammalian lipid phosphate phosphatases (LPPs, or Type 2 phosphatidate phosphohydrolases) constitute a family of enzymes that belongs to a phosphatase superfamily. The LPPs dephosphorylate a variety of bioactive lipid phosphates including phosphatidate, lysophosphatidate, sphingosine 1-phosphate, and ceramide 1-phosphate. Mouse LPP-1 was stably expressed in rat2 fibroblasts to determine its structural and functional properties. Transduced cells showed increased dephosphorylation of exogenous lysophosphatidate. This result is compatible with mutational studies that show the active site of LPP-1 to be located on the external surface of the plasma membrane. Elevated LPP-1 activity attenuated the ability of lysophosphatidate to stimulate mitogen-activated protein kinase (ERK1 and 2) activities and DNA synthesis. It is concluded that one function of LPP-1 is to dephosphorylate exogenous lysophosphatidate, thereby attenuating cell signaling through endothelial cell differentiation gene (EDG) receptors. PMID- 10818445 TI - The role of de novo ceramide synthesis in chemotherapy-induced apoptosis. AB - The de novo pathway of sphingolipid synthesis has been implicated as an alternative to sphingomyelinase activation in generating an apoptotic response through ceramide. A chemotherapy agent was used to activate this pathway in a human T-cell line in order to investigate the role of de novo ceramide synthesis in apoptosis. In data obtained from intact cell radiolabeling studies, it was observed that the first and rate-limiting enzyme in de novo synthesis, serine palmitoyltransferase, is activated and controls the production of ceramide through this pathway. Furthermore, using agents that selectively inhibit ceramide production by this pathway, partial protection from cell death was observed that was independent of caspase activation. These results reveal that serine palmitoyltransferase, an enzyme that controls sphingolipid synthesis for housekeeping functions, is activated during apoptosis and serves to mediate events in this process. PMID- 10818447 TI - Neurobiology of receptor-mediated lysophospholipid signaling. From the first lysophospholipid receptor to roles in nervous system function and development. AB - Identification of the first lysophospholipid receptor, LPA1/Vzg-1, cloned by way of neurobiological analyses on the embryonic cerebral cortex, has led to the realization and demonstration that there exist multiple, homologous LP receptors, including those encoded by a number of orphan receptor genes known as "Edg," all of which are members of the G-protein-coupled receptor (GPCR) superfamily. These receptors interact with apparent high affinity for lysophosphatidic acid (LPA) or sphingosine-1-phosphate (S1P or SPP), and are referred to based upon their functional identity as lysophospholipid receptors: LPA and LPB receptors, respectively, with the expectation that additional subgroups will be identified (i.e., LPC, etc.). Here an update is provided on insights gained from analyses of these receptor genes as they relate to the nervous system, particularly the cerebral cortex, and myelinating cells (oligodendrocytes and Schwann cells). PMID- 10818446 TI - PPAR-gamma-mediated regulation of normal and malignant B lineage cells. AB - Prostaglandins of the E-series stimulate B lymphocytes by enhancing immunoglobulin-class switching and antibody production. Little is known about whether or not other prostaglandins affect B lineage cells and perhaps counterbalance the stimulatory effects of PGE2. PGD2 is a major product of cyclooxygenase in bone marrow and in macrophages, suggesting a role for this lipid product in immunological responses. PGD2 undergoes dehydration to the biologically active prostaglandin 15-deoxy-delta 12,14-PGJ2 (15d-PGJ2) that binds to the nuclear receptor known as peroxisome proliferator-activated receptor gamma (PPAR-gamma). We found that normal mouse B cells and a variety of B lymphoma cells (e.g., 70Z/3, WEHI-231, CH12, and J558) express PPAR-gamma mRNA and the 67 kDa PPAR-gamma protein. 15d-PGJ2 had a dose-dependent antiproliferative/cytotoxic effect on normal and malignant B cells, as shown by 3H-thymidine and MTT assays. Only PPAR-gamma agonists (i.e., thiazolidinediones) mimicked the effect of 15d PGJ2 on B lineage cells, indicating that the mechanism by which 15d-PGJ2 negatively affects B lineage cells involves PPAR-gamma. The mechanism whereby PPAR-gamma agonists induced cytotoxicity is via apoptosis, as shown by Annexin V assay. PPAR-gamma agonists may serve as a counterbalance to the stimulating effects of PGE2, which promotes B-cell differentiation. The use of prostaglandins, such as 15d-PGJ2, and synthetic PPAR-gamma agonists to induce apoptosis in B lineage cells may lead to the development of therapies for fatal PGE2-resistant B lymphomas. PMID- 10818448 TI - EDG receptors as a therapeutic target in the nervous system. AB - EDG receptors are a family of closely related G-protein-coupled receptors, so called since the first family member to be cloned is encoded by an endothelial differentiation gene. Of the six family members identified, five use lysophospholipids as their endogenous ligands. The sixth receptor, EDG-6, remains an orphan. These receptors activate multiple secondary-messenger pathways involving coupling to Gi, Gq/11, and G12/13 trimeric guanine nucleotide-binding proteins and are thought to play an important role in cell growth, development and maintenance, and cytoskeletal-dependent changes. EDG receptors are expressed in most mammalian cells and tissues, each subtype having a distinct distribution pattern, raising the possibility of tissue-specific biological roles that could be explored in drug-discovery programs. In this study the distribution of EDG receptor mRNA within the nervous system has been investigated. As seen in peripheral tissues, these receptors appear to be discretely localized within specific brain regions and cell types. For example, EDG-1, -3, -4 receptors are confined to neuronal cells, EDG-2 receptors to white matter tracts, while EDG-5 receptors appear to be expressed in various cell types, including neuronal cells, white matter tracts, and ependymal cells. EDG-6-receptor mRNA was not detected in the nervous system. Speculation as to the role of these receptors in physiological/pathophysiological processes, particularly those involving cell development, proliferation, maintenance, migration, differentiation, plasticity, and apoptosis can be made from such distribution studies. EDG receptors located in brain neuronal cells might, for example, influence apoptosis and be involved in cell rescue following ischemic damage or during the early stages of progressive neurodegenerative diseases. Those restricted to oligodendrocytes might play a crucial role in myelination and offer a potential target in the treatment of demyelinating diseases, such as multiple sclerosis. In order to explore the role of these receptors, it is necessary to identify selective compounds. To this end we have developed an agonist-induced [35S]GTP gamma S binding assay using an HEK cell line expressing a pertussis-toxin-insensitive human-EDG-2-receptor-rat-Gi alpha 1-fusion protein. Such as assay system overcomes the problems associated with the almost ubiquitous responsiveness of mammalian cells to lysophospholipid. This assay lends itself to high throughput application, opening up the possibility of identifying compounds to further probe the therapeutic potential of EDG receptor manipulation. PMID- 10818449 TI - Lysophosphatidic acid induction of neuronal apoptosis and necrosis. AB - Lysophosphatidic acid (LPA) elicits a unique response in primary hippocampal neurons and sympathetic neuron-like cells, PC12 cells differentiated with nerve growth factor; LPA is cytotoxic. Treatment of rat hippocampal neurons with 50 microM LPA resulted in necrosis, as determined morphologically and by release of lactate dehydrogenase. Lower concentrations of LPA, 0.1, and 1 microM, induced neuronal apoptosis, as assessed by chromatin condensation, annexin V binding, TUNEL staining, and the caspase sensitivity of these events. In addition, 10 and 25 microM LPA induced apoptosis of PC12 cells. In order to define intracellular events associated with this neuronal apoptosis, protective agents were identified. Neurons and PC12 cells were protected against LPA-induced apoptosis by pretreatment with the antioxidant, propyl gallate, or with nitric oxide synthase inhibitors. PC12 cells were protected by insulin and insulin-like growth factor-1 treatment. There is also evidence for mitochondrial participation in LPA mediated apoptosis, including cyclosporin A-mediated protection. Thus, LPA induced neuronal apoptosis is associated with mitochondrial alterations, the generation of reactive oxygen species and nitric oxide, and protection by pretreatment with a serum constituent, insulin-like growth factor 1. PMID- 10818450 TI - Phospholipid growth factors and corneal wound healing. AB - In many tissue types, wound healing involves cell division and migration over and into the wound area to cover and remodel the wound. LPA and other members of the phospholipid lipid growth factor (PLGF) family stimulate many of the activities involved in wound healing. In the rabbit cornea, we have found that keratocytes from wounded corneas have a volume-activated Cl- current activated by LPA and alkenyl-LPA. This current is minimally activated by cyclic PA and SPC, and is not activated by LPA in cells from uninjured corneas. Biochemical examination of PLGFs in aqueous humor and lacrimal fluid before and after wounding identified LPA, alkenyl-GP, PA, and lyso PS, with elevated PLGF activity after wounding. In recent experiments examining human corneal cell lines and cultured cells using RT PCR, we found mRNA for EDG receptors 1-5, with an apparent increase in EDG-3, -4, and -5 following brief SDS application to cell lines, and EDG receptors 2-5 induction in late-passage human corneal epithelial cells. This work points to a significant role for PLGFs in the corneal wound-healing process. PMID- 10818451 TI - LPA as a paracrine mediator of adipocyte growth and function. AB - Adipogenesis corresponds to the recruitment of new adipocytes in adipose tissue, and results from the proliferation/differentiation of preadipocytes. Production of paracrine and autocrine factors by adipocytes plays an important role in adipogenesis. We recently demonstrated the existence of adipocyte production of lysophosphatidic acid (LPA) both in vitro and in situ. This production is modulated by catecholamines via alpha 2-adrenergic receptors. Adipocyte-LPA present in conditioned media increases the growth of a preadipose cell line in culture. This growth is associated with an activation of mitogen-activated protein kinases, and of the focal adhesion kinase. Because of the close proximity of preadipocytes and adipocytes within adipose tissue, adipocyte-LPA could play an important role in autocrine/paracrine control of adipogenesis. PMID- 10818452 TI - Eicosanoid regulation of angiogenesis in human prostate carcinoma and its therapeutic implications. AB - Cancer of the prostate is the most commonly diagnosed cancer in America. There are several lines of evidence implicating the involvement of arachidonate 12 lipoxygenase, an enzyme metabolizing arachidonic acid to form 12(S) hydroxyeicosatetraenoic acid (HETE), in prostate cancer progression. First, as prostate cancer reaches a more advanced stage, the level of 12-lipoxygenase expression is increased. Second, overexpression of 12-lipoxygenase in human prostate cancer cells stimulates angiogenesis and tumor growth. Third, an inhibitor of 12-lipoxygenase has been found effective against metastatic prostate tumor growth, and the inhibition of 12-lipoxygenase is related with the reduction of tumor angiogenesis. Collectively, these studies suggest that 12-lipoxygenase regulates tumor angiogenesis in prostate cancer and that inhibition of 12 lipoxygenase is a novel therapeutic approach for the treatment of prostate cancers. PMID- 10818453 TI - Mechanisms of lysolipid phosphate effects on cellular survival and proliferation. AB - The specificity of cellular effects of lysolipid phosphate (LLP) growth factors is determined by binding to endothelial differentiation gene-encoded G protein coupled receptors (EDG Rs), which transduce diverse proliferative and effector signals. The primary determinants of cellular responses to LLPs are the generative and biodegradative events, which establish steady-state concentrations of each LLP at cell surfaces, and the relative frequency of expression of each EDG R. There are major differences among types of cells in the net effective generation of the LLPs, lysophosphatidic acid (LPA) and sphingosine 1-phosphate (S1P), and in their profile of expression of EDG Rs. The less well characterized secondary determinants of cellular specificity of LLPs are high-affinity binding proteins with carrier and cell-presentation functions, cell-selective regulators of expression of EDG Rs, and cellular factors that govern coupling of EDG Rs to G protein transductional pathways. The roles of components of the LLP-EDG R system in normal physiology and disease processes will be definitively elucidated only after development of animal models with biologically meaningful alterations in genes encoding EDG Rs and the discovery of potent and selective pharmacological probes. PMID- 10818454 TI - Lysophospholipid growth factors in the initiation, progression, metastases, and management of ovarian cancer. AB - Levels of lysophosphatidic acid (LPA) and lysophosphatidylcholine (LPC) are elevated in the plasma and ascites of ovarian cancer patients, but not in most other tumor types. LPA increases cell proliferation, cell survival, resistance to cisplatin, cell shrinkage, and production of vascular endothelial growth factor, urokinase plasminogen activator, and LPA itself in ovarian cancer cells, but not in normal ovarian surface epithelial cells. PSP24 and members of the endothelial differentiation gene (EDG) family (EDG1, EDG2, EDG4, and EDG7) of G protein coupled receptors mediate LPA signaling. Ovarian cancer cell lines do not express EDG1 mRNA, have variable EDG2 mRNA and protein levels, and frequently exhibit levels of EDG4 mRNA and protein, suggesting that EDG4 may contribute to the deleterious effects of LPA in ovarian cancer. In contrast, activation of the EDG2 LPA receptor on ovarian cancer cells may lead to apoptosis and counter the effects of other LPA receptors. Thus, the development of agonists and antagonists for the appropriate spectrum of LPA receptors may alter proliferation, apoptosis, or response to therapy of ovarian cancer cells. Indeed, over 60% of all current drugs target the G protein-coupled family of receptors, making the LPA receptor family a "drugable" target. LPC, although not as thoroughly studied, increases cellular proliferation and mediates multiple other functions through unique signaling pathways. PMID- 10818455 TI - Sphingolipid regulation of female gonadal cell apoptosis. AB - Ceramide and sphingosine-1-phosphate (S1P) are sphingosine-based lipid signaling molecules that have been implicated as key mediators of cellular growth, differentiation, and apoptosis. The cellular response depends on cell type, on the absence or presence of other signals initiated by the same or another stimulus, and on the subcellular location of sphingomyelin hydrolysis leading to ceramide generation. Consistent with mounting evidence implicating components of the sphingomyelin pathway as mediators of cellular life and death in nonreproductive tissues, recent data have indicated that sphingolipid-based signaling events are also prominent features of cellular development and apoptosis in the fetal and postnatal female gonads. This area of investigation represents a new research avenue of considerable significance for both basic biology and clinical medicine because of the massive levels of developmental death that occur normally in the female germ line, especially during gametogenesis, as well as of the central role of oocyte apoptosis in female gonadal failure resulting from pathologic insults. PMID- 10818456 TI - Structure-function analyses of eicosanoid receptors. Physiologic and therapeutic implications. AB - Prostaglandins (PGs) are ubiquitous lipid mediators derived from cyclooxygenase (COX) metabolism of arachidonic acid that exert a broad range of physiologic activities including modulation of inflammation, ovulation, and arterial blood pressure. The physiologic actions of PGs are mediated in part by their interaction with specific G-protein-coupled PG receptors. Eight PG receptors have been cloned, including four for the major COX metabolite, PGE2. The physiologic roles of the PGE2 receptors have been investigated utilizing subtype-selective agonists, localization of receptor mRNA expression, and creation of mice with targeted disruption of PG receptor genes. These analyses have delineated discrete roles for the various PG receptor subtypes. Recent studies on mice lacking the PGE2 EP2 receptor have implicated the PGE2 EP2 receptor subtype in arterial dilatation and salt-sensitive hypertension, and also indicate that this receptor plays a key role in female fertility. The EP2 receptor may thus prove to be a productive target for pharmacological intervention in the treatment of hypertension and infertility. PMID- 10818457 TI - The molecular pharmacology of lysophosphatidate signaling. PMID- 10818458 TI - Evaluation of plasma lysophospholipids for diagnostic significance using electrospray ionization mass spectrometry (ESI-MS) analyses. AB - We previously reported that lysophosphatidic acid (LPA) represents a potential biomarker for ovarian and other gynecologic cancers. To further improve the accuracy and potentially increase the sensitivity and specificity of the assay, we developed an electrospray ionization mass spectrometry (ESI-MS)-based method to analyze LPA and related lysophospholipids. LPA, lysophosphatidylinositol (LPI), lysophosphatidylserine (LPS), and lysophosphatidylcholine (LPC) could be detected with high sensitivity (in low pmol range) using this method. Standard curves were established for quantitative analysis. LPA and closely related lysophospholipids isolated from thin-layer chromatography (TLC) plates were analyzed directly by ESI-MS. This ESI-MS-based assay allows simultaneous detection and quantitation of all different species of LPAs and LPIs in a sample over a range of at least 5-300 pmol. Moreover, this test was at least 50 times more sensitive when a multiple reaction monitoring (MRM) mode was used. Using these protocols in a limited set of analysis, we found that both LPA and LPI were elevated in patients with ovarian cancer. PMID- 10818459 TI - Switching intracellular signaling pathways to study sphingosine 1-phosphate receptors. PMID- 10818460 TI - A novel human G-protein-coupled receptor, EDG7, for lysophosphatidic acid with unsaturated fatty-acid moiety. PMID- 10818461 TI - Quantitative analysis of lysophosphatidic acid in human blood fractions. PMID- 10818462 TI - Topical application of LPA accelerates wound healing. PMID- 10818463 TI - Local and systemic delivery of an aspirin-triggered lipoxin stable analog inhibits neutrophil trafficking. PMID- 10818464 TI - Analysis of the EDG2 receptor based on the structure/activity relationship of LPA. PMID- 10818465 TI - Stimulation of platelets and endothelial cells by mildly oxidized LDL proceeds through activation of lysophosphatidic acid receptors and the Rho/Rho-kinase pathway. Inhibition by lovastatin. PMID- 10818466 TI - Characterization of endogenous and heterologously expressed LPA receptor subtypes. PMID- 10818467 TI - Regulation of expression of EDG family receptors in human prostate cancer cell lines. PMID- 10818468 TI - Proinflammatory signals upregulate COX-2 and increase PGE2 production in biphenotypic B/macrophage cells. PMID- 10818469 TI - Peroxisome proliferator-activated receptor gamma (PPAR-gamma) activation in naive mouse T cells induces cell death. PMID- 10818470 TI - Sphingosine-1-phosphate inhibits haptotactic motility by overproduction of focal adhesion sites in B16 melanoma cells through EDG-induced activation of Rho. AB - We investigated the molecular mechanism by which Sph-1-P affects the FN-dependent haptotactic motility of serum-starved mouse melanoma B16/F10 cells. We found that EDG-5-induced Rho activation followed by enhanced tyrosine phosphorylation of FAK and paxillin, and beta 1-integrin activation leading to overexpression of focal adhesion sites, as well as increment of stress fiber formation, must be the molecular basis of inhibition of haptotactic cell motility by Sph-1-P. PMID- 10818471 TI - Sphingosylphosphorylcholine is a bona fide mediator regulating heart rate. PMID- 10818472 TI - Endothelin-1 stimulates sphingosine kinase in human hepatic stellate cells. A novel role for sphingosine-1-P as a mediator of growth inhibition. PMID- 10818473 TI - Platelet phospholipids decrease vascular endothelial permeability via a novel signaling pathway independent of cAMP/protein kinase A. PMID- 10818474 TI - A novel lipid mediator, cyclic phosphatidic acid (cPA), and its biological functions. PMID- 10818475 TI - Effects of 15-deoxy-delta 12,14-PGJ2 on B lineage cells. PMID- 10818476 TI - Lysophospholipid-induced cell migration. PMID- 10818477 TI - Structural features of EDG1 receptor-ligand complexes revealed by computational modeling and mutagenesis. PMID- 10818478 TI - Effect of sphingosine-1-phosphate and analogues of lysophosphatidic acid on mesangial cell proliferation. PMID- 10818479 TI - 2-arachidonoylglycerol: a possible multifunctional lipid mediator in the nervous and immune systems. PMID- 10818480 TI - Lysophosphatidic acids produced by lysophospholipase D in mammalian serum and body fluid. PMID- 10818481 TI - Acyl and alkyl ether-linked lysophosphatidic acids in the nervous system. PMID- 10818482 TI - Vascular factors in cognitive impairment--where are we now? PMID- 10818483 TI - Role of perivascular cells and myocytes in vascular amyloidosis. AB - Amyloidogenic processing of amyloid-beta precursor protein (APP) by cells of the brain is the major pathologic component of Alzheimer's disease. Amyloid-beta (A beta) is of heterogeneous origin. Perivascular cells of monocyte-macrophage microglial cell lineage produce fibrillar A beta in the wall of capillaries, whereas parenchymal microglial cells produce fibrillar A beta in the parenchyma of gray matter. Fibrillar A beta deposition by perivascular cells lead to endothelial cell degeneration and death, obliteration of affected capillaries, and reduction of the length of the vascular network. These changes cause local ischemia with neuronal degeneration and death. Smooth muscle cells are the source of A beta in the tunica media of parenchymal and leptomeningeal arteries and veins. Fibrillar A beta in the tunica media of leptomeningeal and parenchymal vessels causes degeneration and necrosis of smooth muscle cells and leads to multiple cortical hemorrhages. Smooth muscle cells isolated from blood vessels with amyloid deposits secrete A beta and accumulate nonfibrillar A beta intracellularly. The amyloidogenic processing of APP can be enhanced by apolipoprotein E, reduced by transthyretin, and modulated by several cytokines. PMID- 10818484 TI - Binswanger disease: the history of a silent epidemic. PMID- 10818485 TI - Cascading glia reactions: a common pathomechanism and its differentiated control by cyclic nucleotide signaling. AB - A pathological glia activation, stimulated by inflammatory proteins, beta amyloid, or brain ischemia, is discussed as a common pathogenic factor for progressive nerve cell damage in vascular and Alzheimer dementia. A critical point seems to be reached, if the cytokine-controlled microglial upregulation causes a secondary activation of astrocytes which loose the negative feedback control, are forced to give up their physiological buffering function, and may add to neuronal damage by the release of nitric oxide (NO) and by promoting toxic beta-amyloid formation. A strengthening of the cyclic adenosine-5',3' monophosphate (cAMP) signaling exerted a differential inhibition of the stimulatory cytokines tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 beta (IL-1 beta) released from cultured rat microglia, but maintained the negative feedback signal IL-6; cAMP inhibited also the release of free oxygen radicals (OR) but not of NO. Reinforcement of the NO-induced cyclic guanosine monophosphate (cGMP) increase by blockade of the phosphodiesterase (PDE) subtype 5 with propentofylline counterbalanced the toxic NO action that causes with OR neuronal damage by peroxynitrate formation. In rat cultured astrocytes, a prolonged cAMP elevation favored cell differentiation, the expression of a mature ion channel patter, and an improvement of the extracellular glutamate uptake. Cyclic AMP signaling could be strengthened by PDE blockade and by raising extracellular adenosine, which stimulates A2 receptor-mediated cAMP synthesis. Via an A1 receptor-mediated effect, elevated adenosine was found to overcome a deficient intracellular calcium mobilization resulting from an impaired muscarinic signaling at pathologically decreased acetylcholine concentrations. We suggest that pharmaca, which elevate extracellular adenosine and/or block the degradation of cyclic nucleotides, may be used to counteract glia-related neuronal damage in dementing processes. PMID- 10818486 TI - Linguistic ability in early life and the neuropathology of Alzheimer's disease and cerebrovascular disease. Findings from the Nun Study. AB - Findings from the Nun Study indicate that low linguistic ability in early life has a strong association with dementia and premature death in late life. In the present study, we investigated the relationship of linguistic ability in early life to the neuropathology of Alzheimer's disease and cerebrovascular disease. The analyses were done on a subset of 74 participants in the Nun Study for whom we had handwritten autobiographies completed some time between the ages of 19 and 37 (mean = 23 years). An average of 62 years after writing the autobiographies, when the participants were 78 to 97 years old, they died and their brains were removed for our neuropathologic studies. Linguistic ability in early life was measured by the idea (proposition) density of the autobiographies, i.e., a standard measure of the content of ideas in text samples. Idea density scores from early life had strong inverse correlations with the severity of Alzheimer's disease pathology in the neocortex: Correlations between idea density scores and neurofibrillary tangle counts were -0.59 for the frontal lobe, -0.48 for the temporal lobe, and -0.49 for the parietal lobe (all p values < 0.0001). Idea density scores were unrelated to the severity of atherosclerosis of the major arteries at the base of the brain and to the presence of lacunar and large brain infarcts. Low linguistic ability in early life may reflect suboptimal neurological and cognitive development, which might increase susceptibility to the development of Alzheimer's disease pathology in late life. PMID- 10818487 TI - Cerebrovascular pathology in Alzheimer's disease and leukoaraiosis. AB - A high percentage of patients with Alzheimer's disease (AD) show evidence of white matter degeneration known as leukoaraiosis (LA), which is due to chronic ischemia. We found that the periventricular veins tend to become occluded by multiple layers of collagen in the vessel walls in the elderly. This collagen deposition is particularly excessive in LA lesions. Therefore, it is present in the brains of many AD patients, along with other ischemia-causing cerebrovascular pathology. We found evidence that there is severe loss of oligodendrocytes in LA, due to extensive apoptosis. No evidence of inflammation was found in the LA lesions. In thick celloidin sections of AD brain, we have obtained detailed 3D views of small (early) deposits of amyloid (stained with beta-amyloid antibody) around capillaries (stained with collagen IV antibody). PMID- 10818488 TI - Role of blood vessels in producing pathological changes in the brain with Alzheimer's disease. AB - Vascular factors have been shown to be highly involved in the deposition of the amyloid beta-protein (A beta) in the brain of Alzheimer's disease (AD). However, the detailed mechanism remains unknown. Here, we showed that more numerous deposits of A beta 40 and A beta 42 in the brain were found in AD patients than in controls. Together with evidence of no difference in the level of A beta 40 and A beta 42 in sera between sporadic AD and controls, a certain dysfunction of the blood-brain barrier could induce an abnormal transport of A beta from sera to the parenchyma in AD. In addition, vascular A beta deposits and mature A beta plaques stained by Congo red in AD brains contained more A beta 40 than A beta 42, whereas Congo red-negative immature plaques mainly consisted of A beta 42. Our confocal laser scanning microscopy demonstrated an intimate relationship between A beta 40 and the vascular network. The amount of mature plaques but not that of immature plaques was reportedly correlated with the severity of dementia in AD patients. These results suggest that serum-derived A beta 40 and/or A beta 42 cause A beta 40 deposition in and around blood vessels through unknown but possible mechanisms such as (1) endocytosis of A beta 40, (2) selective transport A beta 40 and A beta 42 into blood vessels and the parenchyma, respectively, and (3) proteolysis of A beta 42 into A beta 40 induced by a putative carboxyl dipeptidase in blood vessels including vascular feet, which is involved in A beta fibrillation and cognitive deterioration in the patients. Therefore, the accumulation of A beta 40 associated with blood vessels may play a critical role in the development of AD. PMID- 10818489 TI - Cerebrovasculature-mediated neuronal cell death. AB - The presence of significant vascular disease in patients with Alzheimer's disease (AD) and the recognition of the ApoE genotype as a risk factor for both coronary disease and AD support an association between AD and vascular disease. It is our hypothesis that brain microvessels contribute to the pathogenesis of AD by producing soluble factors that injure or kill neurons. In this study we report that AD microvessels produce factors that are noxious to neurons and that these vessels can evoke neuronal cell death in vitro. In these experiments, microvessels are isolated from the cerebral cortices of AD patients and non demented elderly and young controls. Microvessels isolated from AD brains produce high levels of a known neurotoxin nitric oxide, compared to vessels from aged matched controls. In addition, we demonstrate a direct neurotoxic effect of AD microvessels when co-cultured with primary rat cerebral cortical neurons. In contrast, vessels from elderly non-demented donors are less lethal, and brain vessels from younger donors are not neurotoxic. Similarly, AD vessels exhibit a dose-dependent toxicity in co-culture with the human neurons. Finally, treatment of AD microvessels with the protein synthesis inhibitor cycloheximide reduces AD vessel neurotoxicity, suggesting that the neurotoxic factor is a protein. These findings suggest that the cerebral microvasculature is a source of factors that can injure neurons and implicate a novel mechanism of vascular-mediated neuronal cell death in AD. PMID- 10818490 TI - Role of aberrant nitric oxide synthase-3 expression in cerebrovascular degeneration and vascular-mediated injury in Alzheimer's disease. AB - Nitric oxide (NO) is an important signaling molecule that is generated through the catalytic activity of nitric oxide synthase (NOS). In the brain, NO mediates neuronal survival, synaptic plasticity, vascular smooth muscle relaxation, and endothelial cell permeability. Previous studies demonstrated aberrant expression of the NOS-III gene in neurons and glial cells in brains with Alzheimer's disease (AD). Since NOS-III is also expressed in vascular cells, and cerebrovascular disease (CVD) frequently complicates the pathology of AD, we investigated the role of NOS-III in relation to CVD in AD. Vasculopathy in AD + CVD was characterized by thickening and hyalinization of the media of small and medium size vessels, variable degrees of beta-amyloid (A beta) deposition, and increased apoptosis of vascular smooth muscle and endothelial cells, particularly involving white matter vessels. These abnormalities were correlated with reduced levels of NOS-III expression in cerebral vessels. Double-labeling studies demonstrated that the low levels of cerebrovascular NOS-III were associated with increased levels of the pro-apoptosis gene product, p53 in smooth muscle and endothelial cells, suggesting a role for altered NOS-III expression in AD-associated vascular degeneration. Constitutively reduced cerebrovascular NOS-III expression and NO production could also lead to cerebral hypoperfusion due to impaired vasodilation responses, and diminished capacity to remove respiratory waste products and toxins from the extracellular space due to reduced capillary permeability. The role for phosphodiesterases as modulators of NOS activity is discussed, as these molecules represent potential therapeutic targets given their cell type and cyclic nucleotide specificities of action. PMID- 10818491 TI - Similar ultrastructural breakdown of cerebrocortical capillaries in Alzheimer's disease, Parkinson's disease, and experimental hypertension. What is the functional link? AB - The brain, as an intensely active organ, is highly dependent on a sufficient nutrient and oxygen availability in order to reach its optimal working capacity. It is well known that the vital supply of energy substrates is provided by the circulatory system, which splits up into a fine, terminal capillary network in target tissues. These capillaries are considered as important sites, since the actual nutrient trafficking takes place through their walls. That is why an intact, preserved structure of the microvessels is crucial to fulfill their function. Since the brain is known to be particularly vulnerable to suboptimal oxygen and glucose delivery, the intact morphology of capillaries is of paramount importance. Several observations have indicated that the cerebral capillary ultrastructure is damaged in Alzheimer's disease (AD). Curiously, the regional cerebral blood flow of AD patients is also significantly lower than in age matched control individuals. Based on these data, it has been suggested that the decreased blood supply and the cerebrovascular alterations contribute to the development of dementia. However, we have observed similar capillary damage in Parkinson's disease patients and chronically hypertensive rats in addition to AD cases, as presented here. These findings indicate that cerebral capillary damage is not exclusive for AD but occurs under other neurodegenerative disorders and hypertension, as well. We hypothesize that ultrastructural abnormalities of cerebral capillaries are causally related to decreased cerebral blood flow and create a condition that favors neurodegenerative mechanisms including the development of dementia. PMID- 10818492 TI - Distribution of amyloid beta 42 in relation to the cerebral microvasculature in an elderly cohort with Alzheimer's disease. AB - Amyloid beta (A beta) deposits and neurofibrillary pathology are characteristic features of Alzheimer's disease (AD). The association of A beta with cerebral vessels is an intriguing feature of AD. While some degree of cerebral A beta angiopathy involving the leptomeninges and intraparenchymal vessels occurs in almost all cases of AD, the proportion of microvessels within a neocortical region containing deposits of A beta peptide is not known. In this study, we examined a cohort of clinically and pathologically evaluated AD cases to assess the percentage of cerebral microvessels in the temporal cortex and parahippocampal gyrus associated with the predominant, A beta 42 form of the peptide. We also assessed whether the distribution and burden of amyloid was related to apolipoprotein E (APOE) genotype. Using double immunostaining methods, we surprisingly found that at least 40% of the microvessels in the two brain regions contained A beta 42 deposits. There was no correlation of such localization with APOE genotype, however, epsilon 4 homozygotes revealed a greater burden of A beta 40. These observations suggest that high proportions of cortical microvessels are associated with A beta 42, which may affect microvascular function. PMID- 10818493 TI - Cerebrovascular smooth muscle cell surface fibrillar A beta. Alteration of the proteolytic environment in the cerebral vessel wall. AB - Cerebrovascular deposition of the amyloid beta-protein (A beta) is a common pathologic event in patients with Alzheimer's disease (AD) and certain related disorders including hereditary cerebral hemorrhage with amyloidosis Dutch-type (HCHWA-D). A beta deposition occurs primarily in the medial layer of the cerebral vessel wall in an assembled fibrillar state. These deposits are associated with several pathological responses including degeneration of the smooth muscle cells in the cerebral vessel wall. Severe cases of cerebrovascular A beta deposition are also accompanied by loss of vessel wall integrity and hemorrhagic stroke. Although the reasons for this pathological consequence are unclear, altered proteolytic mechanisms within the cerebral vessel wall may be involved. Recent studies from our laboratory have shown that cell-surface assembly of A beta into fibrillar structures causes cellular degeneration via an apoptotic pathway and creates an altered proteolytic microenvironment on the cell surface of human cerebrovascular smooth muscle cells (HCSM cells). For example, HCSM cell-surface A beta fibrils serve as a site for tight binding of cell-secreted amyloid beta precursor protein (A beta PP). Since A beta PP is a potent inhibitor of key proteinases of coagulation cascade, its enhanced localization on the A beta fibrils would provide an strong anticoagulant environment. In addition, HCSM cell surface A beta fibrils are potent stimulators of tissue plasminogen activator (tPA) creating a profibrinolytic milieu. Our findings indicate that A beta fibril assembly on the HCSM cell surface causes cellular degeneration and results in both a strong anticoagulant and fibrinolytic environment. Together, these altered proteolytic events could create a setting that is conducive to loss of vessel wall integrity and hemorrhagic stroke. PMID- 10818494 TI - A beta vasoactivity: an inflammatory reaction. AB - Mounting evidence from in vitro and in vivo studies in transgenic mice overproducing beta-amyloid peptides (A beta) suggests that A beta can induce vasoconstriction and decrease cerebral blood flow. In this report, we describe the vasoactive properties of A beta, in particular the enhancement of endothelin 1-induced vasoconstriction and A beta's induction of a long-lasting vasoconstrictive event. Furthermore, we show that low doses (as low as 50 nM) of freshly solubilized A beta similar to those observed in the plasma of patients suffering from Alzheimer's disease are vasoactive. By using various inhibitors and activators of the phospholipase A2 (PLA2)/arachidonic acid (AA) cascade, we demonstrate that A beta vasoactivity is dependent on activation of this intracellular signaling pathway, resulting in stimulation of downstream cyclooxygenase-2 and 5-lipoxygenase, which mediate production of proinflammatory eicosanoids. Taken together, our data show that A beta directly activates an intracellular proinflammatory pathway, which is responsible for its vasoactive properties. PMID- 10818495 TI - Cerebral amyloid angiopathy: accumulation of A beta in interstitial fluid drainage pathways in Alzheimer's disease. AB - Cerebral amyloid angiopathy (CAA) is characterized by the accumulation of beta amyloid (A beta) peptides in the walls of arteries both in the cortex and meninges. Here, we test the hypothesis that CAA results from the progressive accumulation of A beta in the perivascular interstitial fluid drainage pathways of the brain. Experimental studies have shown that interstitial fluid (ISF) from the rat brain flows along periarterial spaces to join the cerebrospinal fluid (CSF) to drain to cervical lymph nodes. Such lymphatic drainage plays a key role in B-cell and T-cell mediated immunity of the brain. Anatomical studies have defined periarterial ISF drainage pathways in the human brain that are homologous with the lymphatic pathways in the rat brain but are largely separate from the CSF. Periarterial channels in the brain in man are in continuity with those of leptomeningeal arteries and can be traced from the brain to the extracranial portions of the internal carotid arteries related to deep cervical lymph nodes. The pattern of deposition of A beta in senile plaques and in CAA suggests that A beta accumulates in pericapillary and periarterial ISF drainage pathways. A beta could accumulate in CAA due to either (i) increased production of A beta, (ii) reduced solubility of A beta peptides, or (iii) impedance of drainage of A beta along periarterial ISF drainage pathways within the brain and leptomeninges due to aging factors in cerebral arteries. Elucidation of factors that reduce elimination of A beta via perivascular drainage pathways may lead to their rectification and to new strategies for treatment of Alzheimer's disease. PMID- 10818496 TI - Traumatic brain injury elevates the Alzheimer's amyloid peptide A beta 42 in human CSF. A possible role for nerve cell injury. AB - The increased risk for Alzheimer's Disease (AD) associated with traumatic brain injury (TBI) suggests that environmental insults may influence the development of this age-related dementia. Recently, we have shown that the levels of the beta amyloid peptide (A beta 1-42) increase in the cerebrospinal fluid (CSF) of patients after severe brain injury and remain elevated for some time after the initial event. The relationships of elevated A beta with markers of blood-brain barrier (BBB) disruption, inflammation, and nerve cell or axonal injury were evaluated in CSF samples taken daily from TBI patients. This analysis reveals that the rise in A beta 1-42 is best correlated with possible markers of neuronal or axonal injury, the cytoskeletal protein tau, neuron-specific enolase (NSE), and apolipoprotein E (ApoE). Similar or better correlations were observed between A beta 1-40 and the three aforementioned markers. These results imply that the degree of brain injury may play a decisive role in determining the levels of A beta 1-42 and A beta 1-40 in the CSF of TBI patients. Inflammation and alterations in BBB may play lesser, but nonetheless significant, roles in determining the A beta level in CSF after brain injury. PMID- 10818497 TI - Prospects for noninvasive imaging of brain amyloid beta in Alzheimer's disease. AB - The brain in patients with Alzheimer's disease (AD) contains large amounts of fibrillary amyloid beta protein. Studies attempting to use levels of amyloid beta protein in plasma, cerebrospinal fluid or skin as diagnostic tests for the disease have not been fruitful. A method for the noninvasive detection of cerebral amyloid beta would be valuable for dementia differential diagnosis, pathophysiology and monitoring of anti-amyloid therapies. Anti-amyloid monoclonal antibody 10H3 has been evaluated as an amyloid-imaging ligand, without success. Important considerations in the development of amyloid-imaging ligands include choice of radiolabel and physical and biological half-lives, route of administration, protein binding, use of control molecules, and imaging techniques. It is important that imaging studies be designed to reflect the slow nature of the process of amyloid deposition. We used a transgenic mouse model overexpressing beta protein precursor (beta PP) to assess the binding of basic fibroblast growth factor (bFGF) and serum amyloid P component (SAP) to amyloid beta (A beta) plaques in mouse brain. Although the binding of these ligands is similar to AD, neither is found endogenously associated with A beta deposits. Because SAP is a component of mouse serum, these findings suggest the blood-brain barrier in transgenic mice is not affected as it is in AD. These findings suggest that the transgenic mouse may be used as a model for evaluation of A beta imaging methods. PMID- 10818498 TI - A newly formed amyloidogenic fragment due to a stop codon mutation causes familial British dementia. AB - Familial British dementia (FBD) is an early-onset autosomal dominant disorder characterized by progressive cognitive impairment, spasticity, and cerebellar ataxia. Hippocampal neurofibrillar degeneration and widespread parenchymal and vascular amyloid deposits are the main neuropathological lesions. Amyloid fibrils are composed of a novel 34 amino acid subunit (ABri) with no sequence identity to any known amyloid molecule. The peptide derives from a larger precursor protein codified by a single gene BRI on chromosome 13. Affected family members have a single base substitution at the stop codon of the BRI gene that generates a longer open-reading frame resulting in a larger precursor protein. The release of the 34 C-terminal amino acids from the mutated precursor originates the ABri amyloid subunit. Our discovery of a new amyloid associated with the development of dementia supports the concept that amyloid peptides may be of primary importance in the initiation of neurodegeneration. PMID- 10818499 TI - Relationship between severe amyloid angiopathy, apolipoprotein E genotype, and vascular lesions in Alzheimer's disease. AB - In this brief review, we aim to describe the complex relationship between cerebral amyloid angiopathy (CAA), apolipoprotein E (ApoE), and cerebrovascular lesions in Alzheimer's disease (AD). First, we review the evidence that CAA is associated with, and may cause, specific types of vascular lesions (VLs). In addition to being a leading cause of lobar hemorrhages in the elderly, CAA has been implicated as a likely cause of small infarcts, microinfarcts, and incomplete infarctions in the deep white matter. We also review the role that ApoE4 (the major genetic risk factor for AD) has in predisposing toward CAA, coronary artery disease, and possibly toward cerebrovascular disease. Last, we provide evidence that the association between CAA and VLs is not a spurious one due to an increase in the ApoE4 genotype. Even within patient groups with the same ApoE genotype (specifically, E4/4 homozygotes and E3/3 homozygotes), our recent analyses have found significant increases in VLs in association with severe CAA. We discuss the implications of this finding as advancing a pathogenic role for severe CAA in producing many of the VLs commonly found in AD cases. PMID- 10818500 TI - Plasma beta-amyloid peptide, transforming growth factor-beta 1, and risk for cerebral amyloid angiopathy. AB - Despite the documented association between apolipoprotein E genotype and cerebral amyloid angiopathy (CAA), a substantial proportion of CAA-related hemorrhages occur in patients without known risks for this disorder. Two other factors implicated in the pathogenesis of CAA are the amyloid-beta peptide (preferentially deposited in vessels as a 40-amino acid species) and the multifunctional cytokine transforming growth factor-beta 1 (a specific promoter of vascular amyloid deposition in transgenic models). We measured plasma concentrations of these factors in a series of 25 patients diagnosed with probable or definite CAA-related hemorrhage and compared them with 21 patients with hemorrhage due to probable hypertensive vasculopathy and 42 elderly control subjects without hemorrhage. We found no differences among the groups in concentrations of the 40- or 42-amino acid species of beta-amyloid or either the active or latent form of transforming growth factor-beta 1. While the data do not exclude important roles for these molecules as risks for CAA, they indicate that plasma measurements are not useful in its diagnosis. PMID- 10818501 TI - The effect of iron and aluminum on transferrin and other serum proteins as revealed by isoelectric focusing gel electrophoresis. PMID- 10818502 TI - A beta vasoactivity in vivo. AB - Bilateral temporoparietal hypoperfusion has been frequently observed early in the Alzheimer's disease (AD) process. The beta-amyloid (A beta) peptide is believed to play a central role in the pathogenesis of AD. In vitro experiments have shown that freshly solubilized A beta enhances constriction of cerebral and peripheral vessels. We proposed that in vivo, A beta would also have vasoactive properties. To test this hypothesis, we intraarterially infused freshly solubilized A beta 1 40 in rats and observed changes in peripheral blood pressure, cerebral blood flow, and cerebrovascular resistance. We found that infusion of A beta in vivo significantly increased the blood pressure in hypotensive rats but not in normotensive and hypertensive rats. Moreover, A beta infusion also resulted in a decreased blood flow and increased vascular resistance specifically in cerebral cortex but not in heart or kidneys. These data suggest that A beta has a direct and specific constrictive effect on cerebral vessels in vivo, which may contribute to the cerebral hypoperfusion observed early in the AD process. PMID- 10818503 TI - Cytochemistry of intraplatelet Ca++ spots as a peripheral marker of age-related brain impairment. PMID- 10818504 TI - Lipoproteins in the central nervous system. AB - Although the synthesis and metabolism of plasma lipoproteins are well characterized, little is known about lipid delivery and clearance within the central nervous system (CNS). Our work has focused on characterizing the lipoprotein particles present in the cerebrospinal fluid (CSF) and the nascent particles secreted by astrocytes. In addition to carrying lipids, we have found that beta-amyloid (A beta) associates with lipoproteins, including the discoidal particles secreted by cultured astrocytes and the spherical lipoproteins found in CSF. We believe that association with lipoproteins provides a means of transport and clearance for A beta. This process may be further influenced by an interaction between A beta and apoprotein E (apoE), the primary protein component of CNS lipoproteins. Specifically, we have investigated the formation and physiologic relevance of a SDS-stable complex between apoE and A beta. In biochemical assays, native apoE2 and E3 (associated with lipid particles) form an SDS-stable complex with A beta that is 20-fold more abundant than the apoE4:A beta complex. In cell culture, native apoE3 but not E4 prevents A beta-induced neurotoxicity by a mechanism dependent on cell surface apoE receptors. In addition, apoE and the inhibition of apoE receptors prevent A beta-induced astrocyte activation. Therefore, we hypothesize that the protection from A beta induced neurotoxicity afforded by apoE3 may result from clearance of the peptide by SDS-stable apoE3:A beta complex formation and uptake by apoE receptors. PMID- 10818505 TI - Apolipoprotein E genotype and cerebral amyloid angiopathy-related hemorrhage. AB - Following the identification of the role of the apolipoprotein E (APOE) gene polymorphism in Alzheimer's disease (AD), this gene was examined in cerebral amyloid angiopathy (CAA). As in AD, the APOE epsilon 4 allele was found to be associated with CAA. Lobar intracerebral hemorrhage is the major clinical manifestation of CAA. Initial studies on a small number of patients with CAA related hemorrhage (CAAH) identified overrepresentation of APOE epsilon 4. However, it became clear that confounding bias from concomitant AD and the need for pathologically confirmed cases of CAAH would also have to be considered. A larger series of pathologically confirmed cases of CAAH, also assessed for AD pathology, found a surprising overrepresentation of the APOE epsilon 2 allele. Because of the association between CAA and AD, it might have been predicted that patients with CAAH would have a low, rather than a high, epsilon 2 frequency. The overrepresentation of APOE epsilon 2 was present both in patients with and without AD, whereas a high epsilon 4 frequency correlated with concomitant AD. Further studies found that overrepresentation of APOE epsilon 2 is specific for CAAH and is not found in intracranial hemorrhages due to other causes. In CAAH, APOE epsilon 2 may interact with putative risk factors for hemorrhage, including antiplatelet/anticoagulant medication, minor head trauma, and hypertension. Several microvascular abnormalities in amyloid-laden blood vessels have been assumed to antedate CAAH and increase its likelihood. APOE epsilon 2 has now been found to be associated with some of these vascular abnormalities, specifically a "double-barrel" appearance and fibrinoid necrosis. The currently favored interpretation is that APOE epsilon 4 enhances deposition of amyloid-beta protein in the walls of cerebral blood vessels, whereas epsilon 2 is a risk factor for hemorrhage from amyloid-laden blood vessels by promoting specific "CAA-associated vasculopathies." PMID- 10818506 TI - Apolipoprotein E, smooth muscle cells and the pathogenesis of cerebral amyloid angiopathy: the potential role of impaired cerebrovascular A beta clearance. AB - Cerebral amyloid angiopathy (CAA) is caused by the deposition of beta-amyloid (A beta) in Alzheimer disease brains. It also occurs isolated, representing a major cause for cerebral hemorrhage in the elderly. The E4 genotype of apolipoprotein E (ApoE) is a risk factor for CAA; however, the molecular mechanism underlying this genetic association is unknown. Various findings suggest that cerebrovascular A beta is derived from the soluble A beta contained in the cortical extracellular space or the cerebrospinal fluid (CSF) that communicates and surrounds small cortical or leptomeningeal vessels. CAA deposits are always intimately associated with smooth muscle cells (SMCs) or SMC-derived pericytes. As we have previously reported, SMCs internalize A beta in vitro via a lipoprotein pathway involving ApoE and the low-density lipoprotein receptor family. Internalized A beta is subsequently located to lysosomes, suggesting its intracellular degradation. We show that A beta is internalized via multiple pathways, because class A and class B scavenger receptors are also colocalized to A beta-containing endosomes in SMCs, and A beta uptake is inhibited by various scavenger receptor antagonists. It has been recently shown for different cell types that the cellular uptake of ApoE is more efficient for the ApoE3 isoform when compared to ApoE4 and that this isoform-specific difference depends on the presence of heparan sulfate proteoglycan (HSPG). HSPG is produced by SMCs and promotes A beta fibrillogenesis. We propose a pathogenetic model of CAA, in which the ApoE- and HSPG-mediated clearance of CSF-derived A beta peptides by SMCs protects the vascular extracellular matrix against critical A beta concentrations. Impairment of this pathway or its reduced efficiency in carriers of the ApoE4 genotype may increase the risk of developing CAA. PMID- 10818507 TI - Amyloid-beta-induced degeneration of human brain pericytes is dependent on the apolipoprotein E genotype. AB - Amyloid-beta (A beta) deposition in cerebral vessels (cerebral amyloid angiopathy, CAA) is accompanied by degeneration of vascular cells, including pericytes and smooth muscle cells. Previous studies indicated that specific A beta protein isoforms are toxic for cultured human brain pericytes and smooth muscle cells. In particular, A beta 1-40 carrying the E22Q mutation, as in hereditary cerebral hemorrhage with amyloidosis of the Dutch type (HCHWA-D), is toxic. We investigated the effects of the A beta-binding protein apolipoprotein E (ApoE) on the toxicity of A beta for cultured human brain pericytes. We compared the toxicity of HCHWA-D A beta 1-40 for pericyte cultures with different ApoE genotypes, studied the accumulation of A beta and ApoE in these different cell cultures, and investigated the effects of exogenous ApoE. Pericyte cultures with an ApoE epsilon 2/epsilon 3 genotype were more resistant to HCHWA-D A beta 1-40 treatment than cultures with a epsilon 3/epsilon 3 or epsilon 3/epsilon 4 genotype. Cell death was highest in cultures homozygous for ApoE epsilon 4. The extent to which both A beta ApoE accumulated at the cell surface was parallel to the degree of toxicity. The addition of purified ApoE resulted in a decrease in cell death. These data suggest that ApoE4 may direct A beta more efficiently than other ApoE isoforms into a pathological interaction with the HBP cell surface. The results of this study are in line with the observations that inheritance of the ApoE epsilon 4 allele increases the risk of developing Alzheimer's disease, and that the ApoE epsilon 2 allele has a relatively protective effect. PMID- 10818508 TI - Earlier age of onset of Alzheimer's disease in patients with both the transferrin C2 and apolipoprotein E-epsilon 4 alleles. AB - The etiology of Alzheimer's disease is now known to be multifactorial. The genetic factors transferrin C2 (TfC2) and apolipoprotein E epsilon 4 (ApoE epsilon 4) have both been associated with Alzheimer's disease (AD). Transferrin is the carrier protein for iron in the blood, while ApoE is involved with the transport and redistribution of lipids. In the present study, the polymerase chain reaction (PCR) method was used to determine the frequency of both TfC2 and ApoE-epsilon 4 in 27 AD patients, 9 vascular dementia (VaD) patients, and 27 controls. Patients were diagnosed according to the criteria as set out in the 4th edition of the Diagnostic and Statistical Manual of the American Psychiatric Association (DSM-IV). The frequency of the TfC2 allele for the AD patients was 24%, while for the VaD patients it was 12.5%, which was not significantly different from the controls at 13%. The frequency of ApoE-epsilon 4 for the AD patients was 44%, for the VaD patients 22%, and controls 17%. Of the 27 AD patients, 8 had both TfC2 and ApoE-epsilon 4. The age of onset of the disease in these 8 patients (51-67 years, mean 60.25) was significantly earlier (p < 0.02) than in the remaining AD patients (49-76 years, mean 66.9). None of the VaD patients had both the TfC2 and the ApoE-epsilon 4 alleles. PMID- 10818509 TI - Inherent abnormalities in energy metabolism in Alzheimer disease. Interaction with cerebrovascular compromise. AB - Alzheimer disease (AD) is a form of the dementia syndrome. AD appears to have a variety of fundamental etiologies that lead to the neuropathological manifestations which define the disease. Patients who are at high risk to develop AD typically show impairments of cerebral metabolic rate in vivo even before they show any evidence of the clinical disease on neuropsychological, electrophysiological, and neuroimaging examinations. Therefore, impairment in energy metabolism in AD can not be attributed to loss of brain substance or to electrophysiological abnormalities. Among the characteristic abnormalities in the AD brain are deficiencies in several enzyme complexes which participate in the mitochondrial oxidation of substrates to yield energy. There include the pyruvate dehydrogenase complex (PDHC), the alpha-ketoglutarate dehydrogenase complex (KGDHC), and Complex IV of the electron transport chain (COX). The deficiency of KGDHC may be due to a mixture of causes including damage by free radicals and perhaps to genetic variation in the DLST gene encoding the core protein of this complex. Inherent impairment of glucose oxidation by the AD brain may reasonably be expected to interact synergistically with an impaired supply of oxygen and glucose to the AD brain, in causing brain damage. These considerations lead to the hypothesis that cerebrovascular compromise and inherent abnormalities in the brain's ability to oxidize substrates can interact to favor the development of AD, in individuals who are genetically predisposed to develop neuritic plaques. PMID- 10818510 TI - Insulin effects on glucose metabolism, memory, and plasma amyloid precursor protein in Alzheimer's disease differ according to apolipoprotein-E genotype. AB - Higher fasting plasma insulin levels and reduced CSF-to-plasma insulin ratios, suggestive of insulin resistance, have been observed in patients with Alzheimer's disease (AD) who do not possess an apolipoprotein E (ApoE)-epsilon 4 allele. Insulin has also been implicated in processing of beta-amyloid and amyloid precursor protein (APP). We examined the effects of intravenous insulin administration while maintaining euglycemia on insulin-mediated glucose disposal, memory, and plasma APP in patients with AD and normal adults of varying ApoE genotypes. AD subjects without an epsilon 4 allele had significantly lower insulin-mediated glucose disposal rates than did AD patients with an epsilon 4 allele (p < 0.03) or than did normal adults without an epsilon 4 allele (p < 0.02). AD subjects without an epsilon 4 allele also showed significant memory facilitation with insulin administration (p < 0.04), whereas the AD-epsilon 4 group did not. Insulin reduced APP levels for AD patients without an ApoE epsilon 4 allele, but raised APP for AD patients with an ApoE epsilon H4 allele These results document ApoE-related differences in insulin metabolism in AD that may relate to disease pathogenesis. PMID- 10818511 TI - Interface between vascular dementia and Alzheimer syndrome. Nosologic redefinition. AB - Vascular dementia is redefined so as to include noninfarct vascular dementia: vascular dementia caused by underlying vascular factors other than cerebral infarction. Data are presented that bring into focus the interface between vascular dementia and Alzheimer syndrome, and the ambiguous transition between multifocality and diffuse or generalized disease. By cross-cutting both stroke and nonstroke vascular groups, arteriosclerosis, abnormal blood pressure, abnormal electrocardiogram, and other vascular factors are implicated in the distal causality of both infarct and noninfarct vascular dementia. PMID- 10818512 TI - Which vascular lesions are of importance in vascular dementia? AB - Most necropsy surveys of dementia have found that vascular disease is second only to Alzheimer's disease as a cause of dementia. Alzheimer's disease and cerebrovascular disease also often coexist. The purpose of the present study was to determine the nature of the cerebrovascular lesions that are most significant in producing dementia. These were analyzed in a group of cases of dementia in which only vascular pathology was present and, in particular, no more than trivial amounts of Alzheimer-type pathology were present. The cerebrovascular lesions in this group of cases were compared with those in a group of stroke cases who were nondemented and a group of elderly cases without stroke or dementia. Severe cribriform change and deep white/grey matter micro-infarcts were significantly more common in the test group than in either of the control groups, whereas single macroscopic infarcts were more common in the stroke control group than either of the other two groups. Thus, microvascular deep white and grey matter lesions, but not macroscopic infarction, were significant in vascular dementia. The results of this study will be discussed in relation to the view that microvascular lesions may also contribute to dementia in subjects with more extensive Alzheimer-type pathology and thus lower the threshold at which Alzheimer-type pathology becomes clinically manifest. PMID- 10818513 TI - Severity of cardiovascular disease, apolipoprotein E genotype, and brain pathology in aging and dementia. AB - Neuropathological lesions, essential for the diagnosis of Alzheimer's disease (AD), such as senile-neuritic plaques (SP/NPs), neurofibrillary tangles (NFTs), the beta-amyloid load (A beta 4) and the load of PHF-tau did not increase with increased severity of cardiovascular disease in 126 clinically demented and 303 nondemented aged individuals. In contrast, the extent of AD lesions was greater in nondemented and demented individuals with the ApoE epsilon 4 allele compared to those without this allele. On the other hand, the extent of vascular lesions currently used for the diagnosis of vascular dementia (VaD) showed correlation with the cardiovascular index, whereas ApoE epsilon 4 allele did not seem to influence the extent of vascular lesions. The calculated CVI showed significant correlation with premortem estimated Hachinski score, and both the CVI and Hachinski score were higher in demented patients with extensive vascular lesions. Our results demonstrate that ApoE epsilon 4 allele, a known risk factor for dementia, indeed influences the extent of Alzheimer's lesions seen in the brain tissue of demented patients and asymptomatic controls. The cardiovascular disease again seems to influence the extent of vascular lesions. PMID- 10818514 TI - Can PET data differentiate Alzheimer's disease from vascular dementia? AB - The present study endeavored to differentiate Alzheimer's disease (AD) from vascular dementia (VaD) by comparing the metabolic and hemodynamic parameters. Positron emission tomographic (PET) studies were carried out in 13 patients with probable AD and 20 patients with VaD. PET findings were not included in the diagnostic criteria of AD or VaD. Using oxygen-15 labeled compounds, cerebral blood flow (CBF), cerebral metabolic rate of oxygen (CMRO2), oxygen extraction fraction (OEF), cerebral blood volume, and vascular transit time (VTT) were measured quantitatively during the resting state. To evaluate vascular reactivity (VR), CBF was also measured during 7% CO2 inhalation. Regional CBF from the parietal cortex positively correlated with the neuropsychological scores in both AD and VaD groups. The typical parietotemporal pattern of hypoperfusion and hypometabolism was observed in the AD group, whereas the frontal lobe including the cingulate and superior frontal gyri were predominantly affected in the VaD group. The occipital cortex was preserved in both groups. A significant increase of the OEF was found in the parietotemporal areas in the AD group. No significant prolongation was seen with VTT. There was a marked difference in VR between the two groups: VR was depleted in the VaD group, whereas VR was normal in the AD group. The increased OEF with preserved vascular reserve seen in AD may implicate participation of a vascular factor in the pathogenesis of AD, possibly at the capillary level. Thus, PET provides important functional information in discriminating AD from VaD by comparing the patterns of hypoperfusion and/or hypometabolism, and in the understanding of the underlying hemodynamic pathophysiology. PMID- 10818515 TI - Limitations of clinical criteria for the diagnosis of vascular dementia in clinical trials. Is a focus on subcortical vascular dementia a solution? AB - Vascular dementia (VaD) includes several different vascular mechanisms and changes in the brain, and has different causes and clinical manifestations. Critical to its conceptualization and diagnosis are definitions of the cognitive syndrome, vascular etiologies, and changes in the brain. Variation in these has resulted in different definitions of VaD, estimates of prevalence, and types and distribution of brain lesions. This definitional heterogeneity may have been a factor for negative results in prior clinical trials on VaD. We propose that the division of VaD into subtypes can identify a more homogeneous group of patients for drug trials. A so-called "subcortical" VaD could incorporate two old clinical entities "Binswanger's disease" and "the lacunar state." Small vessel disease is the primary vascular etiology, lacunar infarcts and ischemic white matter lesions are the primary type of brain lesions, the subcortical areas and frontal connections are the primary location of lesions, and a subcortical syndrome as the primary clinical manifestation. The clinical syndromes are likely more variable, and urgently need to be categorized. Selection of these patients for clinical trials could mainly be based on brain imaging features, where the essential changes and the main aspects of the lesions include extensive ischemic white matter lesions and lacunar infarcts in the deep gray and white matter structures. Subcortical VaD is expected to show a more predictable clinical picture, natural history, outcomes, and treatment responses. PMID- 10818516 TI - CADASIL: hereditary arteriopathy leading to multiple brain infarcts and dementia. AB - Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) often begins with migraine with aura. Recurrent strokes usually appear between 30 and 50 years of age. The arteriopathy develops slowly, resulting in destruction of smooth muscle cells and thickening and fibrosis of the walls of small and medium-sized penetrating arteries with consequent narrowing of the lumen. This impairs cerebral blood flow, visible in PET, and produces characteristic white-matter hyperintensities in T2-weighted MRI on the basis of which CADASIL may be diagnosed well before the first stroke. Multiple lacunar infarcts, mainly in the frontal white matter and basal ganglia, lead to progressive permanent brain damage manifested as cognitive decline and finally as dementia. At present, no specific therapy is available. Infarcts result from thickening and fibrosis of the walls of small and medium-sized penetrating arteries with consequent obliteration and/or thrombosis. Although the symptoms are almost exclusively neurological, the arteriopathy is generalized and diagnosis can be made on the basis of accumulation of pathognomonic basophilic, PAS-positive and in electron microscopy osmiophilic material between degenerating smooth muscle cells in dermal arteries. CADASIL is caused by missense point mutations in the Notch3 gene, which encodes a transmembrane receptor protein with an important signaling function during development. The gene defects lead to either a gain or loss of a cysteine residue in the extracellular N-terminal part of the molecule, most probably causing a conformational and functional alteration. The function of Notch3 in adults and the definite pathogenesis of CADASIL are still unknown, but interestingly its intramembranous proteolytic cleavage may be regulated or implemented by presenilin similarly as cleavage of amyloid precursor protein in Alzheimer's disease. PMID- 10818517 TI - Skin biopsy value and leukoaraiosis. AB - In the field of leukoaraiosis, the identification of CADASIL and its link to Notch 3 mutation has shed light on the pathogenesis of white matter (WM) abnormalities related to small-vessel disease. Since 1993, its systemic vascular involvement allows skin biopsy diagnosis and research on tissues before postmortem examination. We received 160 skin biopsies from patients presenting subcortical dementia, recurrent strokes, behavioral disturbances or migraines, and suspected CADASIL. Almost all the patients lacked the well-known vascular risk factors. The ultrastructural study was systematically carried out looking at the vessel walls and the other components found in skin. In a third, we found endothelial changes, destruction of vascular smooth muscle cells (VSMCs), and characteristic granular osmiophilic material (GOM). In these cases, the genetic analysis confirmed the Notch 3 mutation. Curiously, the skin biopsies from the other two thirds presented marked alterations within the vessel walls. Such changes included destruction of VSMCs, lack of GOM, and replacement of these cells by an extracellular matrix. Frequently, we noticed endothelial pathological changes as well as other tissue impairments. By now, we are able to describe eight different groups of lesions according to either the prevalence of a lesion or the association of different lesions. The skin biopsy ultrastructural study seems to be highly informative given that we can observe vessel lesions and association of impairments in various tissues that might, in part, explain the brain vessel involvement and then the leukoaraiosis and probably some clinical symptoms. Moreover, these vessel lesions often belonged to young people (30-50 years old), and many of them seemed to run in families. These new data associated with early onset of clinical symptoms and leukoaraiosis would be extremely valuable in clarifying the wide field of leucoencephalopathy and might provide genetic research with new issues. PMID- 10818518 TI - Hereditary vascular dementia linked to notch 3 mutations. CADASIL in British families. AB - The most common form of familial vascular dementia is considered to be CADASIL or cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy, which is now also increasingly manifest in the United Kingdom. CADASIL has been previously dubbed as a familial form of Binswanger disease. However, unlike in Binswanger disease CADASIL does not involve hypertension or other risk factors associated with cardiovascular disease. CADASIL appears to be essentially a disorder of the arteries that is linked to single missense mutations in the NOTCH 3 gene locus on chromosome 19. The pathogenesis of the disorder or the genetic mechanism leading to brain infarcts and dementia is not known. The elucidation of the microvascular pathology evident in CADASIL may be an interesting way to delineate effects of defective genes on brain cells from systemic vascular influences. PMID- 10818519 TI - Linear relation between cerebral phosphocreatine concentration and memory capacities during permanent brain vessel occlusions in rats. AB - The present study investigates the interrelation between cerebral energy state and memory capacities in a rat model of stepwise cerebral vessel occlusions. After acute and subchronic permanent vessel occlusions, cortical energy metabolites (ATP, phosphocreatine, ADP, AMP) were detected by high-pressure liquid chromatography (HPLC) analysis, and the effects on learning, memory, and cognitive behavior were evaluated using a hole-board test. The results of the study demonstrated a drastic decrease in energy-rich phosphates by 33% for phosphocreatine and by 44% for ATP after acute vessel occlusions. In addition, rat working and reference memories were strikingly decreased to about 5% of controls. In contrast, two weeks after four-vessel occlusion, the energy state was almost completely restored to control levels. However, a significant decrease in memory capacities was observed in subchronic state. In summary, this study has demonstrated a close linear relationship (p < 0.001) between an impaired cerebral energy state and brain memory dysfunction after acute and permanent cerebral four vessel occlusion. Thus, this animal model of stepwise reduction of the cerebral blood supply may reflect some clinically relevant processes occurring during cerebrovascular and neurodegenerative diseases. PMID- 10818520 TI - Mechanisms of cerebrovascular amyloid deposition. Lessons from mouse models. AB - Cerebrovascular deposition of amyloid is a frequent observation in Alzheimer's disease patients. It can also be detected sporadically in normal aged individuals and is further found in familial diseases linked to specific gene mutations. The source and mechanism of this pathology are still unknown. It has been suggested that amyloidogenic proteins are derived from blood, the vessel wall itself, or from the central nervous system. In this article evidence is reviewed for and against each of these hypotheses, including new data obtained from transgenic mouse models. In APP23 transgenic mice that develop cerebral amyloid angiopathy (CAA) in addition to amyloid plaques, the transport and drainage of neuronally produced amyloid-beta (A beta) seem to be responsible for CAA rather than vascular A beta production or blood uptake. Although a number of mechanisms may contribute to CAA in humans, these results suggest that a neuronal source of A beta is sufficient to induce vascular amyloid deposition. The possibility to cross genetically defined mouse models of CAA with other mutant mice now has the potential to identify molecular mechanisms of CAA. PMID- 10818521 TI - Alzheimer's disease-like cerebrovascular pathology in transforming growth factor beta 1 transgenic mice and functional metabolic correlates. AB - Alzheimer's disease (AD) is frequently associated with cerebrovascular changes, including perivascular astrocytosis, amyloid deposition, and microvascular degeneration, but it is not known whether these pathological changes contribute to functional deficits in AD. To characterize the temporal relationship between amyloid deposition, cerebrovascular abnormalities, and potential functional changes, we studied transgenic mice that express transforming growth factor-beta 1 (TGF-beta 1) at low levels in astrocytes. TGF-beta 1 induced a prominent perivascular astrocytosis, followed by the accumulation of basement membrane proteins in microvessels, thickening of capillary basement membranes, and later, around 6 months of age, deposition of amyloid in cerebral blood vessels. At 9 months of age, various AD-like degenerative alterations were observed in endothelial cells and pericytes. Associated with these morphological changes were changes in regional cerebral glucose utilization. Preliminary results showed that TGF-beta 1 mice had significantly decreased glucose utilization in the mammillary bodies, structures involved in mnemonic and learning processes. Glucose utilization tended to be decreased in several other brain regions as well; however, in the inferior colliculus, it was markedly higher in TGF-beta 1 mice than in controls. We conclude that chronic overproduction of TGF-beta 1 triggers a pathogenic cascade leading to AD-like cerebrovascular amyloidosis, microvascular degeneration, and local alterations in brain metabolic activity. Similar mechanisms may be involved in AD pathogenesis. PMID- 10818522 TI - The role of apolipoprotein E in the deposition of beta-amyloid peptide during ischemia-reperfusion brain injury. A model of early Alzheimer's disease. AB - Transient brain ischemia in the rat produces a stereotyped pattern of selective neuronal degeneration which simulates early Alzheimer's disease (AD) pathology. The aim of the present study was to determine if apolipoprotein E (ApoE) variables are related to alterations in other proteins which play a central role in the pathogenesis of AD; amyloid precursor protein (APP) and beta-amyloid peptide (A beta). The postischemic time course of ApoE and APP and A beta immunoreactivity in brain was examined at survival time from 2 days to 1 year in rats subjected to 10 min cardiac arrest. These data indicate that there are long lasting alterations of ApoE and A beta after brain ischemia. The most likely stimulus for promoting increase of both ApoE and A beta expression are ischemic reperfusion processes. Our data suggest that ApoE modulates the outcome following cerebral ischemia via molecular events in common with AD pathogenesis. We propose that ischemic-reperfusion processes in brain are the fountain-head of a cycle of molecular and cellular events that have neurodegenerative consequences which finally lead to AD. PMID- 10818523 TI - Alterations of Alzheimer's disease in the cholesterol-fed rabbit, including vascular inflammation. Preliminary observations. AB - We determined the levels of endothelial inflammation using MECA-32 antibody and alpha 4 nicotinic receptor subunit densities employing [3H]epibatidine binding in the brains of Alzheimer's disease (AD) patients, cholesterol-fed rabbits, and appropriate controls. We also assessed rabbit brain for beta-amyloid levels and immunohistochemical localization, and for evidence of blood-brain barrier breach using normally-excluded Evans Blue dye. Dietary cholesterol induced a twofold increase in beta-amyloid concentration in rabbit hippocampal cortex, which may be related to the appearance of beta-amyloid immunoreactivity in the neuropil. Epibatidine binding was significantly decreased in AD superior frontal cortex, but unchanged in the superior frontal cortex of cholesterol-fed rabbits. Increased vascular MECA-32 immunoreactivity occurred in AD and cholesterol-fed rabbit brain. Evans Blue dye could be found in the parenchyma of cholesterol-fed rabbits only, and appeared as pockets of dye surrounding small blood vessels. The data suggest that vascular inflammation can lead to breach of the blood-brain barrier, which may produce biochemical derangements in surrounding brain tissue that are conducive to production of beta-amyloid. PMID- 10818525 TI - Vascular endothelium is a site of free radical production and inflammation in areas of neuronal loss in thiamine-deficient brain. AB - Free radical production in vascular endothelial cells and inflammatory responses in perivascular microglia accompany the selective neuronal death induced by TD. Lipid peroxidation and tyrosine nitration occur in neurons within susceptible areas. Thus, region- and cell-specific oxidative stress contributes to selective neurodegeneration during TD. These data are consistent with the hypothesis that in TD, vascular factors constitute a critical part of a cascade of events leading to increases in blood-brain barrier permeability to nonneuronal proteins and iron, leading to inflammation and oxidative stress. Inflammatory cells may release deleterious compounds or cytokines that exacerbate the oxidative damage to metabolically compromised neurons. Similar mechanisms may operate in the pathophysiology of neurodegenerative diseases in which vascular factors, inflammation and oxidative stress are implicated including AD. PMID- 10818524 TI - Animal model of Alzheimer-like vascular pathology and inflammatory reaction. AB - This in vivo animal model of vascular inflammatory reaction facilitates morphologic and hemodynamic analyses of leukocyte-endothelial interaction and can be monitored by video microscopy and electron microscopy. The model has served as a rapid means to explore the deleterious vascular actions and inflammatory response to the cytokines tumor necrosis factor, interleukin-1 and amyloid-beta, as well as the protective effects of superoxide dismutase, estrogen, and cytokine antagonists. PMID- 10818526 TI - Marked hippocampal neuronal damage without motor deficits after mild concussive like brain injury in apolipoprotein E-deficient mice. AB - Of various biological factors, only allele epsilon 4 of apolipoprotein E (apoE, protein; APOE4, gene) has been thus far suggested as a major determinant of genetic risk for sporadic and late-onset familial Alzheimer's disease (AD). Environmental influences such as lack of education, traumatic brain injury, oxidative stress, environmental toxins, hormonal imbalances, and alterations in immune or inflammatory responses may also contribute to the pathogenesis of AD. Thus genetic susceptibility and environmental risk factors may have synergistic effects on the development of AD. The purpose of present report was to assess whether the gene (APOE) and the environmental risk factor (traumatic brain injury) could interact in hippocampal neuronal degeneration. We investigated the histopathological changes of hipoccampal regions after mild concussive-like brain injury without motor deficits in apoE-deficient mice using the recently described novel weight-drop device. Control mice revealed minimal neurodegenerative changes limited to CA2 and CA3, while apoE-deficient mice showed widespread neuronal degeneration throughout hippocampal subfields and part of dentate gyrus. We also observed widespread glial fibrillary acidic protein (GFAP) immunoreactivity throughout the hippocampus, which was more intense in apoE-deficient mice. The results of this study indicate that even very mild traumatic brain injury could result in widespread hippocampal damage in apoE-deficient mice. This again supports the hypothesis that apoE might play a neurotrophic or neuroprotective function in the central nervous system. PMID- 10818527 TI - Cortical cholinergic denervation elicits vascular A beta deposition. AB - Selective destruction of the cholinergic nucleus basalis magnocellularis (nbm) in the rabbit by the p75 neurotrophin receptor (NTR) immunoglobulin G (IgG) complexed to the toxin saporin leads to the deposition of amyloid-beta (A beta) in and around cerebral blood vessels. In some instances, the perivascular A beta resemble the diffuse deposits observed in Alzheimer's disease (AD). We propose that cortical cholinergic deprivation results, among other perturbations, in the loss of vasodilation mediated by acetylcholine. In addition to a dysfunctional cerebral blood flow, alterations in vascular chemistry affecting endothelial and smooth muscle cells may result in cerebral hypoperfusion and a breached blood brain barrier (BBB). The selective removal of the rabbit nbm and A beta accumulation may serve as an important nontransgenic, and more physiological, model for the testing of pharmacological and immunological agents designed to control the deposition and the deleterious effects of A beta in AD. PMID- 10818528 TI - beta-Amyloid excitotoxicity in rat magnocellular nucleus basalis. Effect of cortical deafferentation on cerebral blood flow regulation and implications for Alzheimer's disease. AB - Alzheimer's disease is the most common type of dementia with a still largely unclear etiopathology. One of the factors that may directly contribute to the development and progression of the disorder is the abundant accumulation of beta amyloid peptides (A beta) in senile plaques. In the present account we review coherent in vivo experimental evidence that A beta infusion into the rat magnocellular nucleus basalis (MBN) induces abrupt and persistent behavioral dysfunctions, perturbations of sensory information processing, storage, and retrieval. These substantial behavioral changes are due to the loss of cholinergic neurons in the MBN and their ascending projections to the frontoparietal cortex. Both neuroanatomical and neurochemical observations pin point that infusion of A beta into the rat basal forebrain significantly decreases choline-acetyltransferase and acetylcholinesterase activities and the population of--probably--M2 muscarinic acetylcholine receptors in the cerebral cortex. Neuropharmacological data indicate that A beta toxicity is mediated by an excitotoxic cascade involving blockade of astroglial glutamate uptake, sustained activation of N-methyl-D-aspartate receptors and an overt intracellular Ca2+ influx. These changes are associated with increased nitric oxide synthase activity in cortical target areas that may directly lead to the generation of free radicals. Besides, as microvessels of the neocortex receive direct input from the MBN we assume that the loss of cholinergic innervation and hence that of tonic cholinergic vasoregulation ultimately leads to disturbances of vascular (endothelial) function and nutrient supply that may directly enhance neuronal vulnerability during aging and in Alzheimer's disease. PMID- 10818529 TI - Effect of a memory-enhancing drug, AIT-082, on the level of synaptophysin. AB - Our objective is to study the effect of AIT-082 on the level of synaptophysin in cultured pheochromocytoma (PC12) cells. The drug AIT-082, a unique purine hypoxanthine derivative, is under development for the treatment of Alzheimer's disease (AD). We analyzed synaptophysin protein as an index of synaptic numbers and density and indirectly neuronal transmission. PC12 cells were treated with nerve growth factor (NGF) (50 ng/ml) and/or different doses of AIT-082 (5-50 ng/ml) obtained from NeoTherapeutics, CA. In the western immunoblots of conditioned media and cell lysates, we detected synaptophysin as 36-40 kDa protein bands. When PC12 cells were treated with NGF and samples were analyzed at 24 or 48 hours after treatment, the secretion of synaptophysin was drastically reduced in the conditioned medium. A significant reduction in the intracellular levels of synaptophysin in NGF-treated samples was also noted. By contrast, when PC12 cells were treated with AIT-082, the secretion of synaptophysin was increased in the conditioned medium as compared to the control. There was also a significant increase in the intracellular levels of synaptophysin in AIT-082 treated cultures. NGF treatment resulted in sympathetic neuronal phenotypes in PC12 cells. As it is known that the immunoreactivity of the synaptophysin protein correlates with the density of the synaptic terminal, our results suggest that treatment by AIT-082 could enhance neurotransmitter release at the presynaptic terminal, which may play a role in the improvement of cognition seen in AD subjects. PMID- 10818530 TI - Differentiated cerebrovascular effects of physostigmine and tacrine in cortical areas deafferented from the nucleus basalis magnocellularis suggest involvement of basalocortical projections to microvessels. AB - Cholinesterase inhibitors used to treat Alzheimer's disease according to the principle of cholinergic replacement therapy have proved to be less beneficial than expected. The present study was designed to investigate the cerebrovascular response to physostigmine and tacrine in the experimental model of lesioning of the nucleus basalis magnocellularis (NBM), a model involving a cholinergic deficit. Regional cerebral blood flow was measured by the [14C]iodoantipyrine tissue sampling technique in conscious rats infused with i.v. physostigmine (0.2 mg/kg/h), tacrine (8 mg/kg/h), or saline, 3-5 weeks after unilateral lesion of the NBM with ibotenic acid. Physostigmine and tacrine dose-dependently increased blood flow in most cortical and subcortical regions compared to the control group. However, physostigmine caused smaller blood flow increases in several areas, mostly cortical, of the lesioned compared to the intact hemisphere. The converse was observed with tacrine. A facilitated circulatory response appeared in cortical areas deafferented from the NBM, especially in the frontal cortex. These results provide evidence for distinct NBM-dependent components of the cortical cerebrovascular effects of physostigmine and tacrine. They suggest the involvement of different cellular postsynaptic targets of the NBM. The physostigmine-type effects could involve direct projects onto an inhibitory cortical interneuron supersensitized by deafferentation. This arrangement may explain why physostigmine and perhaps other cholinergic agonists are unable to specifically compensate for a deficit in NBM functioning. The tacrine-type effects presumably involve projections to the microvasculature, including perivascular astrocytes. The neurovascular junction would be sensitized by deafferentation from the NBM. Our data suggest that the regulatory mechanisms of blood flow originating in the NBM might constitute a target of neurodegenerative processes of Alzheimer's disease. PMID- 10818531 TI - Plasma total homocysteine and cognitive performance in a volunteer elderly population. PMID- 10818532 TI - Cardiovascular and other risk factors for Alzheimer's disease and vascular dementia. AB - Factors accelerating cerebral degenerative changes represent potentially modifiable risks for cognitive decline. Putative risks accelerating subtle cognitive decline and dementia were correlated with repeated measures of cerebral atrophy, CT densitometry, perfusions, and cognitive testing among 224 neurologically and cognitively normative aging volunteers. After age 60, cerebral atrophy, ventricular enlargement, polioaraiosis, and leukoaraiosis geometrically increased as perfusions declined. Risks accelerating perfusional decline, cerebral atrophy, polioaraiosis, and leukoaraiosis were: transient ischemic attacks (TIAs), hypertension, smoking, hyperlipidemia, male gender. At age 71.5 +/- 11.9, subtle cognitive decline began, accelerated by TIAs, hypertension, and heart disease. Leukoaraiosis began before cognitive decline. TIAs, hypertension, and hyperlipidemia correlated with vascular dementias. Excessive cortical perfusional decreases and cerebral atrophy correlated with cognitive decline. Family history of neurodegenerative disease correlated with Alzheimer's disease. We concluded that TIAs, hypertension, hyperlipidemia, smoking, and male gender accelerate cerebral degenerative changes, cognitive decline, and dementia. PMID- 10818533 TI - Critically attained threshold of cerebral hypoperfusion: can it cause Alzheimer's disease? AB - After nearly a century of inquiry, the cause of sporadic Alzheimer's disease (AD) remains to be found. On the subject of AD pathogenesis, recent basic and clinical evidence strongly argues in favor of the concept that AD is linked to brain circulatory pathology. This concept, when viewed from many different medical disciplines and from close pre-morbid similarities to vascular dementia, assembles and hypothetically explains most of the key pathologic events associated with the development of AD. These pathologic events are triggered in AD by impaired cerebral perfusion originating in the microvasculature which affects the optimal delivery of glucose and oxygen and results in an energy metabolic breakdown of brain cell biosynthetic and synaptic pathways. We propose that two factors converge to initiate cognitive dysfunction and neurodegeneration as expressed in AD brain: (1) advanced aging, and (2) the presence of a condition that lowers cerebral perfusion. The first factor introduces a normal but potentially deconstructing process that lowers cerebral blood flow in proportion to increased aging, whereas the second factor adds a crucial burden that further lowers brain perfusion to a critical threshold that triggers neuronal metabolic compromise. When age and a condition that lowers cerebral perfusion converge, critically attained threshold of cerebral hypoperfusion (CATCH) results. CATCH is a cyclical and progressive cerebrovascular insufficiency that will destabilize neurons, synapses, neurotransmission, and cognitive ability, eventually evolving into a neurodegenerative process characterized by the formation of senile plaques, neurofibrillary tangles, and amyloid angiopathy. The concept of impaired cerebral perfusion as the cause of this dementia also explains the heterogeneic profile observed in AD patients, because an extensive list of risk factors for AD are also reported to significantly diminish blood flow to the aging brain. PMID- 10818534 TI - The renin angiotensin system and Alzheimer's disease. AB - Recent reports sustain the hypothesis of tight links between vascular and neurodegenerative diseases: associations between atherosclerosis lesions and Alzheimer's disease (AD), increased risk of AD for hypertensive subjects, decreased risk of dementia for elderly treated with hypotensive drugs, and a major impact of apolipoprotein E polymorphism, a protein of the lipid metabolism, on the occurrence of AD. All these results suggest that vascular determinants, both environmental and genetic, may predispose to or speed up dementia. As a major player of vascular homeostasis, the renin angiotensin system (RAS) proteins constitute an interesting source of candidate genes. Among these, the angiotensin I-converting enzyme gene (ACE), a central enzyme of the RAS, presents in its sequence a deletion (D)/insertion (I) polymorphism associated with variations of plasma ACE levels and with the risk of myocardial infarction. We explored the impact of this genetic polymorphism on the risk of cognitive impairment and of dementia in several epidemiological studies. Physiopathological hypotheses suggest a possible involvement of the RAS proteins in the occurrence and evolution of AD. Moreover, although inconsistent, several results of case-control studies tend to suggest that the ACE I/D genetic polymorphism may constitute a genetic susceptibility factor for dementia, reinforcing the hypothesis of a major implication of vascular risk factors in the occurrence of dementia. PMID- 10818535 TI - Neurocardiovascular instability, hypotensive episodes, and MRI lesions in neurodegenerative dementia. AB - We investigated whether carotid sinus hypersensitivity (CSH) and orthostatic hypotension (OH) were associated with a greater severity of hyperintensities on MRI scan in 30 patients with neurodegenerative dementia (17 dementia with Lewy bodies, 13 Alzheimer's disease), who had a detailed evaluation of OH and CSH during active standing and head-up tilt. Patients also underwent a 1.0 Tesla MRI scan, from which hyperintensities were rated on a standardized scale. A blood pressure (BP) drop > 30 mm Hg during carotid sinus massage or active standing was significantly associated with the severity of MRI hyperintensities in the deep white matter (OR 10.0, 95%; CI 1.8-55.7) and in the basal ganglia (OR 11.0, 95%; CI 1.2-99.5) but not in periventricular areas (OR 1.4, 95%; CI 0.3-1.8). Patients with the cardio-inhibitory form of CSH with the largest BP drops were the most at risk. Further longitudinal studies need to investigate the direction of causality to determine whether CSH or OH predispose to MRI hyperintensities and accelerate cognitive decline. PMID- 10818536 TI - beta-Amyloid vasoactivity and proinflammation in microglia can be blocked by cGMP elevating agents. PMID- 10818537 TI - beta-Amyloid fragment 25-35 induces changes in cytosolic free calcium in human platelets. PMID- 10818538 TI - Vascular involvement in cognitive decline and dementia. Epidemiologic evidence from the Rotterdam Study and the Rotterdam Scan Study. PMID- 10818539 TI - Relevance of white matter changes to pre- and poststroke dementia. AB - White matter changes are often associated with stroke, risk factors for stroke, and dementia. From a theoretical point of view, they may be associated with an increased risk of pre- or poststroke dementia because (i) they are linked with subtle cognitive decline, which may add to the consequences of the stroke lesions and of associated Alzheimer pathology; and (ii) they indicate an increased risk of stroke recurrence. The aim of this study was to evaluate the contribution of white matter changes to pre- and poststroke dementia. The relationship between preexisting dementia and white matter changes was evaluated in the Lille stroke dementia cohort. We assessed the cognitive functioning prior to stroke in 202 consecutive patients with ischemic or hemorrhagic stroke, by means of the Informant Questionnaire on Cognitive Decline in the Elderly (IQCODE). We classified in the dementia group patients with IQCODE scores of 104 or more. White matter changes were rated on CT with the Blennow's rating scale. Thirty three of 202 patients were demented before stroke (16.3%; 95% confidence interval: 11.2-21.4); the logistic regression analysis found that female sex, family dementia, white matter changes, and cerebral atrophy were independently associated with prestroke dementia. White matter changes were also associated with an increased risk of poststroke dementia, 2 years after stroke onset. Thus, white matter changes contribute to dementia occurring in stroke patients. PMID- 10818540 TI - Corpus callosum measurement as an in vivo indicator for neocortical neuronal integrity, but not white matter pathology, in Alzheimer's disease. PMID- 10818541 TI - Contrast-enhanced MRI of white matter lesions in patients with blood-brain barrier dysfunction. AB - White matter lesions (WMLs) and blood-brain barrier (BBB) dysfunction are common in dementia. Both conditions may be a consequence of small-vessel disease, in which case the BBB damage could be suspected to be located to the WMLs. Magnetic resonance imaging (MRI) can be used to show WMLs as well as to detect BBB damage when using an intravenous contrast agent, gadolinium. We examined 10 demented patients with WMLs, including 5 cases with BBB (elevated CSF/serum albumin ratios). Results showed no significant changes in MR signal in the WMLs after contrast administration. We conclude that WMLs are not related to BBB damage to such a degree that is detectable with this method and that the elevated CSF albumin might have another origin. PMID- 10818542 TI - The association between white matter lesions on magnetic resonance imaging and noncognitive symptoms. AB - A number of studies have suggested that cerebral changes, particularly deep white matter lesions (WML) visualized on magnetic resonance imaging (MRI), may be involved in the genesis of late life depression. This has been confirmed in a prospective study which also found a relationship between the presence of WML and poor 3-year outcome in elderly depressed subjects. Most studies find these lesions to predominate in frontal lobe and basal ganglia, supporting the hypothesis of "fronto-striatal" dysfunction in depression. To investigate whether WML are associated with mood disturbance in dementia, proton density and T2 weighted images were obtained in 80 subjects with dementia (dementia with Lewy bodies, n = 27; Alzheimer's disease, n = 28; vascular dementia, n = 25) and 26 age-matched normal controls. Periventricular lesions (PVL), white matter lesions (WML), and basal ganglia hyperintensities (BG) were visually rated blind to diagnosis using a semiquantitative scale. Frontal WML were associated with higher depression scores in patients with dementia, implying a common pathophysiology of depression irrespective of diagnosis. Further study of the neurobiological basis of WML is needed. This can best be achieved by serial clinical assessment combined with in vivo and in vitro MRI and neuropathological examination. PMID- 10818543 TI - Neuropathological findings in the very old. Results from the first 101 brains of a population-based longitudinal study of dementing disorders. AB - We report a unique longitudinal epidemiological study of cognitive decline in the elderly population of the city of Cambridge, UK. A population sample of people aged 75 and over was surveyed between 1984-1996 (n = 2,616) and followed 2.4, 6, and 9 years later. CAMDEX diagnostic criteria were used for clinical assessment, and the neuropathological protocol (in 101 cases) was based on the CERAD method, with additional features to allow Braak staging of neurofibrillary pathology. The main findings are of the heterogeneity of lesions to be found in very old populations, and the existence of considerable overlap in the pathologies found in the demented and nondemented. It seems that white matter (ischemic) pallor an amyloid angiopathy, as well as neuritic plaques, neurofibrillary tangles and Lewy body formation are all lesions that increase the likelihood of dementia. PMID- 10818544 TI - Leukoaraiosis at presentation and disease progression during follow-up in histologically confirmed cases of dementia. PMID- 10818545 TI - Vascular actions of estrogen and Alzheimer's disease. AB - Women are two to three times more likely to develop late-onset Alzheimer's disease (AD) than age-matched men. A large number of observational reports and a few randomized clinical trials have indicated that estrogen replacement therapy (ERT) may retard the development and severity of dementia in postmenopausal women. A chronic inflammatory reaction mediated by abnormal deposition of proteins such as amyloid-beta (A beta) is central to the pathology of AD. We investigated the effect of low doses of conjugated estrogen (Premarin) in an animal model of A beta-induced vascular disruption and inflammatory reaction. Estrogen prevented vascular deposition of A beta, endothelial and vessel wall disruption with plasma leakage, platelet and mast cell activation, and characteristic features of an inflammatory reaction: adhesion and transmigration of leukocytes. The beneficial effect was lost when estrogen treatment was discontinued. This novel protective effect of estrogen against A beta-induced vascular dysfunction may contribute to the therapeutic efficacy of estrogen in AD and coronary vascular disease. PMID- 10818546 TI - Subcortical vascular dementia as a specific target for clinical trials. AB - Vascular cognitive impairment is considered the second most common form of mental deterioration in the elderly after degenerative dementias. Therapeutic approaches to vascular dementia mainly rely on the identification and treatment of risk factors. A number of drugs have also been tested with the aim of improving or slowing cognitive decline in patients affected by various forms of cerebrovascular disease. Most of these trials have yielded unsatisfactory results. We hypothesize that some of these failures depend on the inclusion of patients with pathophysiologically heterogeneous types of vascular cognitive decline. In this paper, we review some of the most important trials that tested drugs with a preventive or therapeutic aim in vascular dementia patients. Preliminary results suggest that some beneficial effects can be detected only when the trial population is homogeneous on a clinical and pathogenic basis. In particular, subcortical vascular dementia, a form with a rather univocal clinical, radiological, and pathological picture, seems a particularly apt choice as a target for future clinical studies. At present, only one therapeutic trial is being conducted in patients affected by this specific form of vascular dementia. PMID- 10818548 TI - Glial modulating and neurotrophic properties of propentofylline and its application to Alzheimer's disease and vascular dementia. PMID- 10818547 TI - The diagnosis of "mixed" dementia in the Consortium for the Investigation of Vascular Impairment of Cognition (CIVIC). AB - If vascular risk factors are risk for Alzheimer's disease (AD), and if "pure" vascular dementia (VaD) is less common than has been thought, what do we make of the diagnosis of mixed dementia? We report characteristics of those with mixed dementia in a prospective, seven center, clinic-based Canadian study. Of 1,008 patients, 372 were diagnosed with AD, 149 with vascular cognitive impairment (VCI) including 76 with mixed AD/VaD, and 82 with other types of dementia. The mean age of patients with mixed AD/VaD was 78.0 +/- 7.6 years; 49% were female. These proportions differed significantly between dementia diagnosis subgroup (p < 0.001) showing a trend which is evident in all comparisons--AD/VaD patients fall in between AD and VaD. Vascular risk factors were present significantly more often in mixed AD/VaD than in AD (p < 0.001). More mixed AD/VaD (20%) than AD patients (4%) had focal signs, compared with 38% of those with vascular dementia and 12% with other types of dementia. Between the initial clinical diagnosis and the final diagnosis (which utilized neuroimaging and neuropsychological data) AD/VaD was the least stable diagnosis. Neuroimaging of ischemic lesions was the most common reason for reassignment from AD to the mixed AD/VaD diagnosis (17 cases). These data suggest that an operational definition of mixed AD/VaD can be proposed on presentation and clinical/radiographic findings, but indifferent to vascular risk factors. The concept of mixed dementia should be extended to include vascular dementia in combination with dementias, other than Alzheimer's disease. PMID- 10818549 TI - Investigating the natural course and treatment of vascular dementia and Alzheimer's disease. Parallel study populations in two randomized, placebo controlled trials. PMID- 10818550 TI - Preliminary results from an MRI/CT-based database for vascular dementia and Alzheimer's disease. PMID- 10818551 TI - Alzheimer's disease and vascular dementia. Some points of confluence. AB - The lack of biologic markers for Alzheimer's disease and vascular dementia, the controversy regarding the definition of vascular dementia, and the new evidence of vascular risk factors for Alzheimer's disease suggest that the traditional differentiation between Alzheimer's disease and vascular dementia is no longer very clear. We believe that both vascular and degenerative mechanisms contribute to the development of dementia, especially in very old age. The question of whether they are two independent parallel processes or interacting pathologies needs to be clarified. PMID- 10818552 TI - Identification and psychic closure. A dynamic structuralist approach of the psyche AB - In this paper, the emergence of psychic structures is conceived within a dynamic structuralist framework, developed in dialogue with the semiotic and dynamic viewpoint on levels from evolutionary systems theory. If psychic structures emerge, it is because they are particular kinds of organizationally closed systems that interpret the variety from below (the biological level) for the level above (the social level). The mechanism of psychic closure is presented as the key mechanism for the constitution of psychic structures, and is interpreted in terms of identification: an identificatory judgment is a self-referential, closing judgment that involves a reinterpretation of the level below (the organic body) and is addressed to the level above (the fellow human being). The way in which identification has worked determines the kinds of psychic structures that are formed and also leads to different kinds of signifying practices. Different types of identification are discussed and a brief presentation of clinical arguments serves to illustrate the determinative role of identification in the formation of psychic structures. PMID- 10818553 TI - Doing time. The emergence of irreversibility AB - By considering an enterprise to be a system of agents that observe and construct theories about themselves immediately raises issues of closure. These in turn pose questions about the identity and evolution of that which is exhibiting such closure. We address these questions by assigning enterprises to a class of systems whose models are triply articulated. The existential articulation provides an account of the possible behaviors of the enterprise's agents and of their interoperation. The referential articulation specifies outcomes that its agents are required to satisfy. The deontic articulation imposes constraints on the composition of the other two articulations that are sufficient to ensure that the enterprise effectively implements its specified requirements. Any of these articulations may be under-determined in that they admit more than one elaboration. The behavioral closure of an enterprise is a kind of composition (formally, a category theoretic limit construction) of its three articulations. If the enterprise is its own observer, then the articulations are its models of itself. The enterprise has many opportunities for error in constructing this model. In particular, it may find that it cannot choose among its under determined articulations in such a way that their composition is internally consistent. Such errors necessitate changes to its model, which may be denoted as steps in an irreversible trajectory through a space of such models. This approach seems to provide a conceptual bridge across the gulf between systems theory and psychoanalysis, and has provided valuable insights into strategy formulation within large enterprises. PMID- 10818554 TI - Regenerative process in life and mind. AB - The functional organization of the nervous system is discussed from the standpoint of organizational closure and regenerative process in order to draw parallels between life and mind. Living organization entails continual regeneration of material parts and functional relations (self-production). Similarly, dynamic stability of informational states in brains may entail coherent self-regenerating patterns of neural signals. If mind is the functional organization of the nervous system, then mental states can be seen as switchings between alternative sets of stable, self-regenerative neural signal productions. In networks of neurons, signaling resonances can be created through recurrent, reentrant neural circuits that are organized to implement a heterarchy of correlational operations. Neural representations are dynamically built-up through an interplay between externally-impressed, incoming sensory signals and internally-generated circulating signals to form pattern-resonances. Semiotic aspects of resonance states involve semantic sensori-motor linkages to and through the external environment and pragmatic linkages to evaluative mechanisms that implement internal goal states. It is hypothesized that coherent regenerative signaling may be an organizational requirement for a material system to support conscious awareness. In this view general anesthetics and seizures abolish awareness by temporarily disrupting the organizational coherence of regenerative neural signaling. PMID- 10818555 TI - A classification of closure concepts AB - Closure has been used in various ways. I propose a classification based on the specification hierarchy formalism, wherein ideas are represented as subclasses, or restrictions, of other ideas, beginning with the most generally applicable concept and working inward to more particular applications. This exercise is part of a revival of nineteenth century natural philosophy. The classification also represents a self-referential developmental system in which the classification itself emerges from the evolutionary process it describes. Since one form of closure builds upon a previously attained form, development traces a loss of evolutionary potential even as each closure gives access into a new, emergent, realm. PMID- 10818556 TI - Conceptual closure. How memories are woven into an interconnected worldview. AB - This paper describes a tentative model for how discrete memories transform into an interconnected conceptual network, or worldview, wherein relationships between memories are forged by way of abstractions. The model draws on Kauffman's theory of how an information-evolving system can emerge through the formation and closure of an autocatalytic network. Here, the information units are not catalytic molecules, but memories and abstractions, and the process that connects them is not catalysis but reminding events (i.e., one memory evokes another). The result is a worldview that both structures, and is structured by, self-triggered streams of thought. PMID- 10818557 TI - Importance of diversity. Reconciling natural selection and noncompetitive processes. AB - To better understand selection processes in evolutionary systems (ecological to economic to social to artificial systems), the origins and role of diversity are examined in two systems that show increased group functionality (better performance, efficiency, robustness, adaptability, stability, etc.). Diversity was chosen as a clarifying concept, because it appears to have been largely ignored, or misunderstood. One system is a model of group selection within an ecosystem. The other is the group solution of a sequential problem using self organizing dynamics in the absence of any selection. A comparison of the two systems show that while diversity is essential to both, improvement by natural selection is derived from consuming diversity, whereas improvement by noncompetitive self-organization is decreased by any reduction in diversity. The resulting perspective is that natural selection is a mechanism that increases the functionality of the individual (or groups within a larger system); noncompetitive self-organization of the system, without need for selection, increases the functionality of the whole above that of the individual or group. The two extreme roles for diversity are reconciled if natural selection is not strongly expressed in these systems--"survival of the fittest" becomes "survival of the adequate"--so that noncompetitive processes can occur. The resulting view of a mature ecosystem is an elastic web of interactions in which natural selection is dormant or retains the status quo. The processes of natural selection for individual or group improvement are activated only if environment changes are sufficient to break the elastic interconnections, as might occur in punctuated equilibria. PMID- 10818558 TI - Levels of control and closure in complex semiotic systems AB - It is natural to advance closures as atomic processes of universal evolution, and to analyze this concept specifically. Real complex systems like organisms and complex mechanisms cannot exist at either extreme of complete closure or lack of closure, nevertheless we should consider the properties of closures in general, the introduction of boundaries, a corresponding stability, the establishment of system autonomy and identity, and thereby the introduction of emergent new system of potentially new types. Our focus should move from simple physical closure of common objects and classical self-organizing systems to semiotically closed systems that maintain cyclic relations of perception, interpretation, decision, and action with their environments. Thus, issues arise concerning the use and interpretation of symbols, representations, and/or internal models (whether explicit or implicit) by the system; and the syntactic, semantic, and pragmatic relations among the sign tokens, their interpretations, and their use or function for the systems in question. Primitive semiotic closures are hypothesized as equivalent to simple control systems, and in turn equivalent to simple organisms. This leads us directly to the grand hierarchical control theories of Turchin, Powers, and Albus, which provide an explicit mechanism for the formation of new levels within complex semiotically closed systems. PMID- 10818559 TI - Complexity IX. Closure over the organization of a scientific truth. AB - The specificity of human knowledge allow one to construct specific truths about human behavior. A structural notation and language for describing a complex hierarchically organized biological system was developed for the explicit purpose of analyzing the origins of health and disease. A specific application of these concepts to a specific patient (such as an individual suffering from the heritable disease, sickle cell anemia) requires a systematic formulation of a scientific truth. No universal law is applicable. The value of a clinical truth for the patient, as well as for the physician and society, is substantial. This value has moral, ethical, and legal weight. Both physics and chemistry use a universal external invariant reference system. Human beings and other living organisms function by an internal reference system that is neither invariant nor universal. In order to address the complexity of scientific truths within living systems, a mathematical graph is constructed from observations, descriptions, and symbolizations of the relevant human scientific activities and is placed in mutual coreference with three philosophical theories of truth. Consistency within the referencing relations of the graphic object creates an image of complex truths. When mapped over degrees of internal organization, the structural consistencies can form hierarchically transitive relations (many-to-one) creating redundancies that confirm one-another. Both structural and dynamic information can be composed within the graphic framework. The redundancies intrinsic to the degrees of organization notation, semantics, and syntax augment one another in the search for scientific truth. The degree of certitude emerging from structural implications increases in relation to the number of hierarchical degrees of organization invoked to represent the various behaviors of complex systems (as illustrated by the sickle cell anemia example). The successful synthesis of the complex image (or complex simple) of a scientific truth approaches the Heideggerian notion of identity in the sense that A = A and A is A. PMID- 10818560 TI - Closure, open systems, and the modeling imperative AB - Natural systems cannot be closed to the environment. At the same time there is a necessity for closure in order to build the system. It is this quint-essential tension between openness and closure that drives systems to unfold into further stages or levels of growth and development. In other words, the emergence of organization in natural systems is a result of cycles of openness and closure. There are two distinct and complementary ways by which a system will carry over closure while involved in a process of expansion across the environment. These two ways need to be expressed in any formal representation: (1) within a level this will be by means of transitive closure, which is additive; and (2) between levels (i.e., from one level to the next higher level) this requires algebraic closure, which is multiplicative. The former expresses space closure, whereas the latter expresses topological or time closure. The conjunction of these two closures generates a hierarchy of levels. Prior to, and outside of, the system lies semantic closure. PMID- 10818561 TI - Opening up closure. Semiotics across scales AB - The dynamic emergence of new levels of organization in complex systems is related to the semiotic reorganization of discrete/continuous variety at the level below as continuous/discrete meaning for the level above. In this view both the semiotic and the dynamic closure of system levels is reopened to allow the development and evolution of greater complexity. PMID- 10818562 TI - Closure, identity, and the emergence of formal causation AB - The aim of this paper is to characterize a type of causality relevant to study the closure of complex systems that we call formal causation. By this term we understand the existence of a new (not materially inherent) causal relation among constituents, generated through an autonomous process of closure. Once a certain level of organization is reached, material systems can generate internal constraints that, through recursive processes, construct their own identity. We study two different forms of closure: closure in dissipative systems and closure in template self-replication. Finally, these two forms merge and bring forth a new one: informational closure, We show how complex forms of organization are based on informational closure, which is an explicit, recorded type of formal causation allowing a functional articulation between individual organizations and larger, collective and historical (meta)organizations. PMID- 10818563 TI - Varieties of chemical closure. Three kinds of coherence observed in chemical systems, and their relation to societal integration AB - Covalent bonding within chemical molecules and the internal electronic structure of atoms involve closure of phase relations in electronic wave functions, as suggested de Broglie by many years ago. The structures of crystals involving positive and negative ions can be understood in terms of replication of unit cells that may be classified in terms of symmetry. The main principle involved in crystal symmetry can be understood by examining possible patterns in decorative borders. A more widely applicable type of chemical closure occurs in oscillating reactions (dissipative structures), in which an autocatalytic process is balanced by some exit reaction. As is the case for the other types of chemical coherence, the number of distinct types of oscillating reactions is rather small. Otherwise puzzling aspects of human social and organizational behavior may be clarified by analogy with chemical oscillating reactions. PMID- 10818564 TI - Closure in the emergence and evolution of life. Multiple discourses or one? AB - Evolutionary theory formulated in terms of complex systems dynamics shows interesting convergences with the approaches of developmental systems theory and biosemiotics, especially when applied to the problem of the origin of life. Although starting from difference conceptual assumptions, all three approaches agree on the importance of closure in the form of semipermeable chemical and informational boundaries and a more circumscribed role for DNA. PMID- 10818565 TI - The scientific origin of life. Considerations on the evolution of information, leading to an alternative proposal for explaining the origin of the cell, a semantically closed system. AB - We hypothesize that the origin of life, that is, the origin of the first cell, cannot be explained by natural selection among self-replicating molecules, as is done by the RNA-world hypothesis. To circumvent the chicken and egg problem associated with semantic closure of the cell--no replication of information molecules (nucleotide strands) without functional enzymes, no functional enzymes without encoding in information molecules--a prebiotic evolutionary process is proposed that, from the informational point of view, must somehow have resembled the current scientific process. The cell was the outcome of interactions of a complex premetabolic community, with information molecules that were devoid of self-replicative properties. In a comparable manner, scientific progress is possible, essentially because of interaction between a complex cultural society and permanent information carriers like printed matter. This may eventually lead to self-replicating technology in which semantic closure occurs anew. Explaining the origin of life as a scientific process might provide a unifying theory for the evolution of information, wherebye at two moments symbolization/encoding of interactions into permanent information occurred: at one moment that of chemical interaction and at another moment that of animal behavior interaction. In one event this encoding led to autonomously duplicating chemistry (the cell), an event that possibly may be one of the outcomes of current scientific progress. PMID- 10818566 TI - The Rosetta Stone of stability. AB - A number of proposals to augment the classical theory of natural selection turn on the need to consider the role of neutral genetic modifications, as well as on the role of patterns of physicochemically stable substructures. There are similarities between this opposition and that within cognitive science, where the dynamic and computational approaches emphasize complementary aspects of perceptual-motor development. This paper summarizes the relevant arguments of the various schools of evolutionary and cognitive theory, as well as the various calls that have been made for unification or complementarity of explanations. An extended concept of stability is then proposed, which bridges the gap between thermodynamic equilibrium and semantic closure. PMID- 10818567 TI - From autopoiesis to semantic closure. AB - This article addresses the question of providing an adequate mathematical formulation for the concepts of autopoiesis and closure under efficient cause. What is required is metaphorically equivalent to reducing the act of writing to a set of mathematical equations, habitually effected by a human mathematician, within the ongoing function of the system itself. This, in turn, raises the question of the relationship between autopoiesis and semantics. The hypothesis suggested is that whereas semantics clearly requires autopoiesis, it may be also be the case that autopoiesis itself can only be materially realized in a system that is characterized by a semantic dimension. PMID- 10818568 TI - Remarks on autocatalysis and autopoiesis. AB - The notions of collective autocatalysis and of autopoiesis are clearly related; equally clearly, they are not quite the same. The purpose of this paper is to try to clarify the relationship. Specifically I suggest that autopoiesis can be at least roughly characterized as collective autocatalysis plus spatial individuation. Although some mechanism of spatial confinement or concentration is probably necessary to the effective operation of any collectively autocatalytic reaction network, autopoiesis requires, in addition, that the mechanism for maintaining this confinement should itself be a product of the reaction network- and should thus (?) be capable of separating or individuating otherwise identically organized networks. I suggest an informal heuristic test to discriminate the (merely) collectively autocatalytic from the (properly) autopoietic. Finally, in the light of this, I review a variety of published abstract or model systems, Alchemy, alpha-universes, Tierra, and SCL. PMID- 10818569 TI - Code-duality and the epistemic cut. AB - Howard Pattee has claimed that an epistemic cut separates the world from observers and therefore from organisms. The epistemic cut imputes a linguistic mode of operation to living systems. Among evolutionary systems-theorists on the other hand there seems to be a tendency to consider living systems as just one among many other kinds of evolutionary systems that, in principle, can all be described on the basis of thermodynamics or infodynamics. This paper searches a third position, in which the epistemic cut position is defended without adopting Pattee's distinction between a dynamic and a linguistic mode. It is observed that even the dynamic mode in living systems is always a semiotic mode although index- and analog-coded rather than symbolic and digitally coded. The analog-coded messages corresponds to a kind of tacit knowledge hidden in macromolecular structure and shape (e.g., molecular complementarity), and in organismic architecture and communication, that is, in the semiotic interactions of the body. PMID- 10818570 TI - Closure, function, emergence, semiosis, and life: the same idea? Reflections on the concrete and the abstract in theoretical biology. AB - In this note epistemological problems in general theories about living systems are considered; in particular, the question of hidden connections between different areas of experience, such as folk biology and scientific biology, and hidden connections between central concepts of theoretical biology, such as function, semiosis, closure, and life. PMID- 10818571 TI - Complementarity and closure AB - This paper compares two different accounts of closure to explain biological organization: as an organization enabled by an informational control on dynamics (Pattee) and as an internal closure of a dynamic nature (Varela). The two of them adopt different versions of a principle of complementarity to relate symbolic and dynamic explanations. PMID- 10818572 TI - Syntactic autonomy. Why there is no autonomy without symbols and how self organizing systems might evolve them. AB - Two different types of agency are discussed that are based on dynamically coherent and incoherent couplings with an environment, respectively. I propose that until a private syntax (syntactic autonomy) is discovered by dynamically coherent agents, there are no significant or interesting types of closure or autonomy. When syntactic autonomy is established, then, because of a process of description-based selected self-organization, open-ended evolution is enabled. At this stage, in addition to dynamics, agents depend on localized, symbolic memory, thus adding a level of dynamic incoherence to their interaction with the environment. Furthermore, it is the appearance of syntactic autonomy that enables much more interesting types of closures among agents sharing the same syntax. To investigate how we can study the emergence of syntax from dynamic systems, experiments with cellular automata leading to emergent computation that solves nontrivial tasks are discussed. RNA editing is also mentioned as a process that may have been used to obtain a primordial biological code necessary for open ended evolution. PMID- 10818573 TI - Genomes and form. The case for teleomorphic recursivity. AB - The genotype-phenotype (genome-form) distinction is considered by many to be fundamental to modern evolutionary thinking. Indeed, the premises that: DNA solely constitutes the genotype; that the phenotype is a transient product of the genotype, with the latter not only describing, but also implementing the construction of the former; and that the constructed materials and systems of the cell have no impact on the genotype, have become dogmas. Yet a vast body of data from molecular genetics reveals that cellular systems, directly and indirectly, alter the genome. Some of these data are reviewed. Proteins can influence mutations along the chromosomes, heritably modify the information content of DNA sequences, and, in some instances, reorganize the germline or somatic genome via DNA engineering pathways. These data suggest that the constructed (proteins, chromatin arrays, and metabolic pathways) has an important role in shaping the descriptor. Insofar as it is biochemically possible for states adopted by cellular structures to be stabilized and eventually memorized by engineering chromosomes, semantic closure can be transcended--meaning can be transferred from the domain of form to the genome, and this presumably ongoing process is termed teleomorphic recursivity. Throughout the paper, I implicitly argue that the genome-form partition is strictly a formal one, with no deeply material basis. PMID- 10818574 TI - Fundamental and modal processes in the evolution of natural systems. AB - This paper offers a brief sketch of the evolution of natural systems within the context of cosmic evolution. The argument, conducted from an energy stance but without recourse to the mathematical apparatus of quantum physics, distinguishes the processes by which energy is transformed from the processes in which it is distributed in a local context. The main focus is on the energetic stratification of natural systems, its causes, and its consequences. A brief coda points to some implications of this treatment. PMID- 10818575 TI - Closure and anticlosure in the realm of quantum gravity. Why evolution needs no origin AB - Dynamic systems with suitable nonlinearities yield self-organizing behavior. The evolution continues until the relationship among the components becomes self consistent; that is, until it reaches closure. Disruptions of closure that allow for continued change are also characteristic of biological evolution. Are the requisite nonlinearities add-ons that give an essentially linear world the appearance of circularity, or do they have their origin in the underlying physics of the universe? The picture developed here fits to the latter view. PMID- 10818576 TI - The nature of the organism. Life has a life of its own. AB - The question of closure in biological systems is central to understanding the origins of the biological variation and complexity upon which various forms of selection act. Much of evolutionary theory, especially in the second half of the twentieth century, is concerned with the consequences of environmental selection acting on bio-diversity, but neglects questions of the origin of that diversity. This has permitted us to act as if an explanation of consequences was the ultimate explanation in biology. However, Darwin understood that evolution was both information driven and information constrained. The link between evolutionary constraints and closure can be profitably explored by starting with Darwin's notion of the primary of "the nature of the organism" over "the nature of the conditions" articulated in the sixth edition of Origin of Species. Contemporary ideas of self-organization, emergence, complexity, and inherent (developmental and phylogenetic) constraints can be seen as an elaboration and refinement of Darwin's views if we adopt the following perspective: (1) information is cheap, not costly, to produce, but may have costly consequences; and (2) information is produced by systems that are informationally closed but remain thermodynamically open. PMID- 10818577 TI - Ontic closure and the hierarchy of scale AB - The Newtonian, universalist world view is incompatible with the notion of ontological indeterminacy. A hierarchical, or granular perspective, however, reveals how the consequences of pure chance, at scales for removed from those at which the event occurs, can be mitigated. PMID- 10818578 TI - Active motion, communicative aggregations, and the spatial closure of Umwelt. AB - On the basis of a simple model of movable organisms that are supplied by semiotic force of attraction or repulsion, several general features of spatial behavior are demonstrated: (1) the stochastic spatial distribution of actively moving organisms is unstable; (2) simple or complex aggregations of organisms appear as a result of active motion; and (3) the ability for active motion ties the organisms with place. These results show that models that apply an internalist approach can considerably simplify the theory of spatial behavior of organic systems. Explanations based on the effects of Darwinian fitness may not be necessary for understanding the origin of biological aggregations. PMID- 10818579 TI - Autonomy and process closure as the basis for functionality AB - Most accounts of functionality are based in etiology, either design or selection. In this paper I give an account of function as serving autonomy, which is the closure of self-maintaining processes, including those interacting with the environment. Autonomy is inherently dynamic, being based entirely on interacting processes, whose organization constitutes the integrity of the autonomous system. The etiological account focuses on external factors, either intentions in design or outcomes in selection. It ignores any but idiosyncratic organizational requirements within the biological entity and in its interactions with its environment, even though these may play a central role in the functionality of the trait in question. In particular, there is no simple relation between adaptation on the etiological account and adaptability, a higher order emergent trait that plays a central role in behavior and the evolutionary and developmental genesis of intelligence. I propose redefining adaptiveness in terms of autonomy. The definition naturally extends to adaptability and focuses on the organizational character of adaptiveness, forcing attention on this central biological characteristic, which is easily ignored in the etiological account. The result is a much richer account of both adaptation and selection. PMID- 10818580 TI - The organism and the open system. Ervin Bauer and Ludwig von Bertalanffy. AB - In this historical treatise two biological-system theories, formulated in the 1920s and 1930s, are roughly sketched. The first part discusses the concept of a thermodynamically open system, as coined by the Russian pathologist Ervin Bauer (1890-1942). Like Bertalanffy, Bauer wanted to prove the specificity of the biological sciences against physics. To achieve this, he postulated the necessity to formulate specific laws of motion which are valid for living matter alone. In the second part of the paper, the organismic-system theory of the Austrian Canadian philosopher and biologist Ludwig von Bertalanffy (1901-1972) is outlined. The focus of this theory relied on the process dynamics that is inherent inside an organismic system. Both theories exemplify closure models for a living organism from a methodical point of view that distinguishes these earlier models from semantic closure, developed by Howard Pattee as an epistemic clue in solving the enigma of living phenomena. The objective here is to disclose the essential differences between these closure conceptions. To encourage further research on closure, the essay concludes with a few questions concerning clarification of the term. PMID- 10818581 TI - Organizational closure and conceptual coherence AB - This paper reviews ideas developed by the late Gordon Pask as part of this conversation theory (CT). CT uses theories of the dynamics of complex, self organizing systems, in conjunction with models of conceptual structures, in order to give an account of conceptual coherence (for example, of a theory or a belief system) as a form of organizational closure. In Pask's own terms, CT is concerned both with the kinematics of knowledge structures and the kinetics of knowing and coming to know. The main features of modelling conceptual structures and processes used by Pask are presented. We continue by presenting a summary two cycle model of learning, aimed to capture some of Pask's key insights with respect to conceptual coherence and the organizational closure of conceptual systems. Parallels are drawn with other work in epistemology, classic cybernetic studies of self-organization, and the concept of autopoiesis. The two-cycle model is then applied recursively to generate learning cycles and conceptual structures at different levels of abstraction, as a contribution to the work of Pask on the topology of thought. Finally, the model is applied reflexively. That is, its own form is considered as a topic for conversation and conceptualization. Carrying out such a reflection provides a coherent way of characterizing epistemological limits, while retaining a clear sense of there being an (in principle) unlimited praxeology of awareness. PMID- 10818582 TI - Spontaneous order, autocatakinetic closure, and the development of space-time AB - Over its 4.6 billion year history, the time-dependent behavior of planet Earth, from the origin and emergence of life to the explosive globalization of human culture today, shows the progressive and accelerating production of increasingly more highly ordered dynamic states. Understanding our place as both productions and producers in this rapidly accelerating global becoming is a requisite step to the meaningful grounding of virtually every other discipline, most particularly those disciplines relating to the endeavors and activities of humans themselves. Recent advances in the study of spontaneous ordering provide both a minimal ontological framework required for causally addressing such systems, and the nomological basis for understanding the ubiquitous or universal generic nature of such ordering itself. This paper briefly outlines the main points. PMID- 10818583 TI - The formal nature of emergent biological organization and its implications for understandings of closure. AB - This paper shows how recognition of biological form, of which the dynamics of closure is an integral part, is mandated by research on autopoietic systems, self organization, evolutionary theory, and on topics in a variety of other areas, including infant and child development. It shows how a "matter pure and simple" (a mechanical concept of nature) is inimical to veridical understandings and explanations of emergent organization from the level of cells to the level of intact organisms--animate forms. By means of an analysis and discussion of writings by prominent researchers in diverse fields, a brief inquiry into neuron firing, and a consideration of intrinsic dynamics and primal animation, this paper shows that a principle of motion or animation informs biological nature. In other words, the fundamentally dynamic character of biological form at all levels exemplifies the kinetic character of living matter. The final section of this paper pinpoints implications for closure, emphasizing the need for an acknowledgement of the dynamics of closure and the need for theoretical and biological reformulations of living systems that incorporate that dynamics. It exemplifies the implication in each instance with reference to authors represented in this volume. PMID- 10818584 TI - The internalist stance. A linguistic practice enclosing dynamics AB - Natural dynamics, as manifested in evolutionary processes, refer to material bodies in movement in the present progressive mode. Any interacting material body in the present progressive mode must be sentient to others because there can be no global agency coordinating it to others in a globally synchronous manner. The internalist perspective, or the worm's eye view, referring primarily to the present progressive mode, renders local material bodies, large or small, subject to an inevitable inconsistency among local representations of neighborhood events registered in the local present perfect tense. Any sentient material body experiencing this inconsistency subsequently transforms itself into an inconsistency-free representation. The descriptive scheme unique to the internalist stance is internal and dynamic in the sense that it constantly strives to update constituent local representations, attempting to eliminate any inconsistencies residing within antecedent local representations. Compared to external descriptions of invariable universals grounded upon the Cartesian epistemic split, which are complementary to dynamics, internal description serves as a linguistic means of embodying natural dynamics even without recourse to the notion called forces. This makes our language powerful enough to enclose natural dynamics of material bodies in the empirical domain. PMID- 10818585 TI - Circulating DNA. What we know and what we need to learn. PMID- 10818586 TI - Quantitative aspects of plasma/serum DNA in cancer patients. PMID- 10818587 TI - An overview on the isolation and analysis of circulating tumor DNA in plasma and serum. AB - Molecular biology has brought about a new dimension to our understanding and analysis of human cancer. Diagnosis and prognosis of cancer have increasingly benefited from the use of molecular markers and molecular approaches. Although the description of circulating tumor DNA in the blood has a long history, the clinical implications of its presence require a large body of well-controlled and stringently performed large research and clinical tests. This chapter has provided an overview of the essential elements for the preparation and analysis of circulating DNA in the plasma or serum. We hope that the basic approaches and tips summarized here will serve as a foundation for the rigorous and comprehensive molecular analysis of circulating nucleic acids in the years to come. PMID- 10818588 TI - A review of studies on the detection of mutated KRAS2 sequences as tumor markers in plasma/serum of patients with gastrointestinal cancer. PMID- 10818589 TI - Factors influencing the detection of mutant K-ras in the serum of patients with colorectal cancer. PMID- 10818590 TI - Diagnostic value of K-ras mutations in serum of pancreatic cancer patients. PMID- 10818591 TI - Plasma DNA K-ras mutations in patients with gastrointestinal malignancies. PMID- 10818592 TI - Serum mutant K-ras in the colorectal adenoma-to-carcinoma sequence. Implications for diagnosis, postoperative follow-up, and early detection of recurrent disease. PMID- 10818593 TI - Enrichment methods for mutation detection. PMID- 10818594 TI - Mutation detection in K-ras, BRCA1, BRCA2, and p53 using PCR/LDR and a universal DNA microarray. AB - We have developed a multiplex PCR/ligase detection reaction (PCR/LDR) that combines high sensitivity with the ability to simultaneously detect hundreds of mutations in a single-tube reaction. To enable us to rapidly assay large numbers of samples, we have linked this mutation detection scheme with analysis on a Universal DNA microarray. We have successfully applied this approach to characterize K-ras and p53 mutations in DNA derived from undissected colon tumors. The sensitivity of the assay has also facilitated detection of low frequency mutations in BRCA1 and BRCA2 in pooled samples of DNA; thus, PCR/LDR can rapidly screen large numbers of DNA samples required for population studies. PMID- 10818595 TI - p53 and APC mutations are detectable in the plasma and serum of patients with colorectal cancer (CRC) or adenomas. AB - BACKGROUND: A variety of tumor-related DNA has been detected in the plasma and serum of cancer patients, including RAS, the BCL2-IgH transgene, and p53. It is not known whether the APC gene, which is frequently mutated in colorectal cancer (CRC), can be identified in the plasma of CRC patients. Similarly, it is unknown whether another tumor suppressor gene altered in CRC, p53, is detectable in people with precursor lesions to CRC. MATERIALS AND METHODS: The plasma of 240 colonoscopy patients was tested for mutations at two frequently altered sites (codons 175 and 248). A restriction enzyme-mediated enrichment assay was used to detect these mutants. We also tested tumor tissue from 8 patients with CRC for mutations in the mutation cluster region of the APC gene using direct DNA sequencing. Corresponding plasma from any positive patient was then similarly sequenced. RESULTS: Three plasma p53 mutations were identified. Two of these patients had adenomas at biopsy, and 1 had a hyperplastic polyp. All were tested for the same p53 mutations, and 1 of the adenomas was positive. One of the 8 CRC patients had a 5-base-pair deletion in the cancer and the same deletion was detectable in that patient's plasma, although at an amount that we estimate at 1 5% of the total APC DNA present. CONCLUSIONS: APC mutations are detectable in the plasma of CRC patients. p53 mutants are detectable in the plasma of colorectal adenoma patients. These may have important implications for cancer screening and diagnosis in the large intestine and suggest that all malignant and premalignant DNA alterations from the colon are identifiable in the blood. PMID- 10818596 TI - Profile of proteins complexed with circulating DNA of a lung cancer patient. PMID- 10818597 TI - Microsatellite analysis. Applications and pitfalls. PMID- 10818598 TI - Laser-fluorescence microsatellite analysis and new results in microsatellite analysis of plasma/serum DNA of cancer patients. PMID- 10818599 TI - Detection of tumor genetic alterations of bladder carcinomas in body fluids depends on sample treatment before DNA isolation. PMID- 10818600 TI - Detection of microsatellite alterations in the DNA isolated from tumor cells and from plasma DNA of patients with lung cancer. AB - In this paper, we show that the same panel of three microsatellite markers is useful for the detection of alterations in the DNA of tumor cells and plasma from patients diagnosed with SCLC and NSCLC. In 31% of the SCLC patients, we detected a microsatellite alteration(s) or LOH in at least one locus. In the group of patients diagnosed with NSCLC, a microsatellite alteration or LOH was detected in at least one locus in 33% of the patients. In all but 2 patients, the identical alteration observed in the DNA from tumor cells was also detected in the DNA isolated from blood plasma. This work confirms the results described by other groups and it extends the diagnostic possibilities of finding tumor cell-specific DNA alterations also in the DNA freely circulating in plasma and serum of patients with cancer. PMID- 10818601 TI - Loss of heterozygosity at the ACTBP2 locus in lung cancer detected on Elchrom precast Spreadex gels. AB - One of the most important goals in the continuous struggle against cancer is early detection of the disease by a simple method that can be used in routine laboratory practice. According to several recent reports, early detection may be achieved by monitoring alterations at several microsatellite loci containing tri- and tetranucleotide repeats. The method relies on comparison of the intensity of allele bands in cancer tissue, plasma, and lymphocytes of a patient. Microsatellite instability is typically detected by labeling the DNA with a radioactive probe or a fluorescent dye followed by electrophoresis on 20-40-cm long denaturing polyacrylamide gels. In this study, we show that loss of heterozygosity at the ACTBP2 locus in lung cancer patients can be detected using the Elchrom electrophoresis system and just 4-cm-long precast Spreadex gels. PMID- 10818602 TI - Clinical significance of circulating DNA microsatellite markers in plasma of melanoma patients. PMID- 10818603 TI - Plasma cell-free Epstein-Barr virus DNA quantitation in patients with nasopharyngeal carcinoma. Correlation with clinical staging. PMID- 10818604 TI - Tumor-derived epigenetic changes in the plasma and serum of liver cancer patients. Implications for cancer detection and monitoring. PMID- 10818605 TI - Detection of clone-specific immunoglobulin heavy chain CDR3 PCR products in serum and plasma of patients with B cell malignancies. PMID- 10818606 TI - RT-PCR-based detection of occult disseminated tumor cells in peripheral blood and bone marrow of patients with solid tumors. An overview. AB - Despite recent progress in early detection and local curative therapy, patients with primary epithelial cancer quite frequently relapse with incurable metastasis. Early disseminated tumor cells that may be seminal for distant failure and are undetectable by current diagnostic methods have been identified by immunocytochemical techniques in bone marrow of cancer patients using monoclonal antibodies against cytokeratins. Recently, promising new molecular approaches, namely, reverse transcriptase--polymerase chain reaction (RT-PCR) assays, have been suggested as a potential technique for the detection of minimal residual tumor burden by targeting mRNA transcribed from epithelial genes in bone marrow, peripheral blood, or lymph nodes. Several studies using RT-PCR thus far indicate a highly sensitive and specific staging tool, although the prognostic value is still controversial. However, limitations may arise from ectopic expression of marker mRNA in hematopoietic cells and deficient expression in circulating tumor cells. The present review focuses on the relevant literature and demonstrates the range of current applications of RT-PCR-based assays for detecting disseminated tumor cells in peripheral blood and bone marrow of patients with solid tumors. We will both summarize technical evaluations of published molecular approaches and discuss the widely disparate results on PCR findings in clinical studies. PMID- 10818607 TI - Cellular- versus extracellular-based assays. Comparing utility in DNA and RNA molecular marker assessment. PMID- 10818608 TI - DNA release by line-1 (L1) retrotransposon. Could it be possible? AB - We have verified the presence of line-1 retrotransposon (L1) in plasma DNA in 15/17 brain tumor (glioma) patients and in 6/6 healthy people by applying PCR amplification of part of the L1 5' end. The same samples were separately amplified for K-ras. Results suggested that L1 sequences are circulating throughout the body. We hypothesized the participation of transposable elements such as L1 in a putative DNA release mechanism. PMID- 10818609 TI - Detection of DNA in the plasma of septic patients. AB - Small amounts of plasma-free DNA have been observed both in healthy individuals and in patients with various diseases such as systemic lupus erythematosus, rheumatoid arthritis, viral hepatitis, and cancer. This communication demonstrates that septic patients also release DNA in plasma. After DNA extraction from plasma, exon 1 of the K-ras gene was amplified by PCR and products were analyzed by dot-blot hybridization. Plasmas from polytraumatic patients and control healthy individuals were used for comparisons with septic patients. Our results show that septic patients present DNA in their plasma. As far as we know, this is the first evidence of circulating DNA in septic patients. PMID- 10818610 TI - Fetal DNA in maternal plasma. AB - Recently, cell-free fetal DNA has been found in maternal plasma and serum. This discovery opens up a new field of investigation and provides an easily accessible source of fetal genetic material for prenatal diagnosis. Prenatal diagnostic applications of fetal DNA in maternal plasma include the investigation of sex linked disorders and fetal rhesus D status determination. Cell-free fetal DNA has been found to be present in much higher fractional concentrations than fetal nucleated cells in maternal blood. The concentration of fetal DNA increases throughout pregnancy, with a sharp rise towards the end of gestation. Abnormally high levels of cell-free DNA have been found in pregnancies complicated by preeclampsia and preterm labor, an observation that has potential diagnostic and pathophysiologic implications. Much remains to be learned regarding the mechanisms of production and clearance of maternal plasma fetal DNA. It is hoped that the eagerly awaited answers to these and other questions may ultimately enhance our understanding of the fetomaternal relationship. PMID- 10818611 TI - Multiplex and real-time quantitative PCR on fetal DNA in maternal plasma. A comparison with fetal cells isolated from maternal blood. AB - Fetal DNA has recently been detected in maternal plasma by PCR and has shown promise for the prenatal determination of fetal sex or rhesus D. In order to obtain the maximum amount of information from this fetal genetic material, we have devised a sensitive multiplex PCR method to permit simultaneous analysis for both the SRY locus and the rhesus D gene. Our studies show that this technique is very sensitive and specific. In the 22 cases from rhesus D negative women examined, we were able to determine both fetal genotypes correctly. In the parallel enrichment for fetal cells, fetal erythroblasts were only detected in 14 of the 19 cases. Our data also indicate that fetal DNA from rhesus D positive fetuses is present in maternal plasma even after prophylactic anti-D treatment. Furthermore, since fetal cells have been reported to be elevated in pregnancies with aneuploid fetuses, we have quantified the amount of fetal DNA present in the maternal plasma of 10 such affected pregnancies by real-time PCR. Our results indicate that fetal DNA is elevated under such circumstances when compared to gestationally matched normal pregnancies (mean of 7% in aneuploid samples versus 3.5% in normal pregnancies). These results indicate that the quantification of fetal DNA in maternal plasma may be an additional screening tool for pregnancies at risk of bearing an aneuploid fetus. PMID- 10818612 TI - Determination of fetal RhD status by maternal plasma DNA analysis. PMID- 10818613 TI - Fetal DNA analysis from maternal plasma and nucleated blood cells. PMID- 10818614 TI - The origin and mechanism of circulating DNA. PMID- 10818615 TI - New hypotheses on the material nature of horizontally mobile genes. AB - The evolutionary history of organisms is often assumed to be recorded in the structure of important molecules, such as DNA sequences. Whereas the structure of these molecules does sometimes affirm other evidence of ancestry, like fossil records, it sometimes does not. Horizontal gene transfer can distort perceptions of ancestry. Determining the impact of horizontal gene transfer on evolution has been limited by the crude tools available to detect it. Physical and genetic vectors are now known to conduct genes between organisms, even between biological kingdoms of organisms. The effects are being noticed in important molecules preserved in the genomes of organisms. This article will review the systematic bias in using molecular morphology, like DNA sequences, to infer ancestry and how this bias is the unavoidable result of the way that experimental genetics itself evolved. We present the novel hypothesis that genes usually called epigenes, like methylation patterns and prions, are infectiously transferred, sometimes using DNA as a vector, but not as a gene. PMID- 10818616 TI - Evolutionary Perspectives on Human Reproductive Behavior. Proceedings of a colloquium. New York City, New York, USA. Spring 1999. PMID- 10818617 TI - The relationship between the theory of evolution and the social sciences, particularly psychology. AB - The application of the theory of evolution to human social behavior has, along with some illumination, produced friction that occasionally bursts into flame. In this paper we will examine the relationship between the theory of evolution and the social sciences, psychology in particular. We will identify some of the sources of friction between proponents and opponents of applying evolutionary theory to the social sciences, and we will suggest that listening carefully to both sides in the debate points the way to an enriched understanding of human social behavior. PMID- 10818618 TI - Evolutionary psychology: counting babies or studying information-processing mechanisms. AB - Evolutionary psychology focuses on the study of adaptations. Its practitioners put little credence in the study of reproductive success in recent and current environments, and argue for an information-processing, cost-benefit conception of adaptation. Because ancestral and current environments differ, it is necessary to distinguish between innate and operational adaptations and between concurrently contingent and developmentally contingent behaviors. These distinctions lead to an evolutionary classification of behaviors into true pathologies, pseudopathologies, quasinormal behaviors, and adaptive-culturally-variable behaviors. I argue that a complete study of the functioning of a behavioral adaptation involves modeling ancestral selection pressures, cross-cultural research, experimental studies of mental processes, and studies of the proximate biological correlates of information-processing adaptations. Finally, I claim that evolutionary psychology can help us avoid making both naturalistic and moralistic fallacies. PMID- 10818619 TI - Desires in human mating. AB - This chapter traces theoretical and empirical progress in the study of human mating over the past few decades. Early pre-evolutionary formulations proposed that men and women were identical in their mating motivations. Most were simplistic, typically postulating a single motive for mating: the search for similarity, equity, or complementarity. Given the large sex differences in human reproductive biology, notably women bearing the burdens of internal fertilization and a greater obligatory parental investment, it would be extraordinarily unlikely that evolution by selection would fail to forge sex-differentiated mating strategies. Empirical research over the past 15 years has robustly confirmed evolutionary predictions in the domains of desire for sexual variety, the importance of fertility cues, and the importance of resource-provisioning. Recent work has revealed a hidden side of women's sexuality--a desire for extra pair partners and the conditions under which this desire is expressed. We now have the theoretical and empirical outlines of an evolutionary formulation of human mating strategies. PMID- 10818620 TI - Human sexual selection, good genes, and special design. AB - Evolutionary psychology seeks to understand the functional design underlying psychological processes and behavior. Theories of selection pressures developed within evolutionary biology are key components of this understanding. Because past selection pressures responsible for current design cannot be directly observed, theoretical understandings of the psychological processes and behavior must be inferred. The most important epistemological concept within evolutionary psychology is that of special design-evidence that a feature exhibits specificity, efficiency, and economy for producing a particular beneficial effect. A variety of sexual-selection processes have been proposed to account for aspects of human mating. These processes are not mutually exclusive. More than one may account for aspects of human mating. A core task of evolutionary psychology within this domain is to identify which processes account for which phenomena. I have attempted to illustrate how the search for special design is central to this endeavor. PMID- 10818621 TI - Mental traits as fitness indicators. Expanding evolutionary psychology's adaptationism. AB - According to most evolutionary psychologists, human psychological adaptations can be recognized by criteria such as high efficiency, high complexity, high modularity, low phenotypic variance, low genotypic variance, low heritability, universality across cultures, and universality across individuals. These criteria are appropriate for adaptations that have been shaped through stabilizing selection for survival utility. However, they are often inappropriate for adaptations that have been shaped by sexual selection through mate choice as reliable signals of heritable fitness. If some psychological adaptations evolved as sexually selected fitness indicators of this type, we should expect them to violate many standard criteria used by evolutionary psychology to distinguish adaptations from non-adaptations. This paper addresses the problems raised by new developments in sexual selection theory and animal signaling theory for evolutionary psychology's adaptationism. It suggests that our adaptationist criteria must recognize two typical kinds of psychological adaptations: naturally selected survival mechanisms and sexually selected fitness indicators. PMID- 10818622 TI - The optimal number of fathers. Evolution, demography, and history in the shaping of female mate preferences. AB - Around the world polygynous marriage (one man, several women) is vastly more common than polyandrous marriage (one woman, several men), and women tend to be more cautious about entering into sexual relationships than men are. Such patterns are often assumed to reflect essential differences between the sexes. However, the same dichotomy between "ardent" males and "coy" females is not found in other primates. Furthermore, under a range of circumstances females enhance their reproductive success by mating with multiple partners and use polyandrous mating (soliciting copulations from several or more males) to circumvent male imposed costs on their free choice of mates. The existence of one-male mating systems does not prove that females "naturally" gravitate to them. Typically monandrous (copulating with just one partner) mating systems are maintained by one male excluding rivals or by other circumstances that distort female options. As with many other animals, primate females (including women) can benefit reproductively from polyandrous matings. Understanding this takes us beyond narrow research programs intent on demonstrating "universal" differences between the sexes, and allows us to study females as flexible and opportunistic individuals who confront recurring reproductive dilemmas and tradeoffs within a world of shifting options. PMID- 10818623 TI - The other "closest living relative". How bonobos (Pan paniscus) challenge traditional assumptions about females, dominance, intra- and intersexual interactions, and hominid evolution. AB - Chimpanzee (Pan troglodytes) societies are typically characterized as physically aggressive, male-bonded and male-dominated. Their close relatives, the bonobos (Pan paniscus), differ in startling and significant ways. For instance, female bonobos bond with one another, form coalitions, and dominate males. A pattern of reluctance to consider, let alone acknowledge, female dominance in bonobos exists, however. Because both species are equally "man's" closest relative, the bonobo social system complicates models of human evolution that have historically been based upon referents that are male and chimpanzee-like. The bonobo evidence suggests that models of human evolution must be reformulated such that they also accommodate: real and meaningful female bonds; the possibility of systematic female dominance over males; female mating strategies which encompass extra-group paternities; hunting and meat distribution by females; the importance of the sharing of plant foods; affinitive inter-community interactions; males that do not stalk and attack and are not territorial; and flexible social relationships in which philopatry does not necessarily predict bonding pattern. PMID- 10818624 TI - The elements of a scientific theory of self-deception. AB - An evolutionary theory of self-deception--the active misrepresentation of reality to the conscious mind--suggests that there may be multiple sources of self deception in our own species, with important interactions between them. Self deception (along with internal conflict and fragmentation) may serve to improve deception of others; this may include denial of ongoing deception, self inflation, ego-biased social theory, false narratives of intention, and a conscious mind that operates via denial and projection to create a self-serving world. Self-deception may also result from internal representations of the voices of significant others, including parents, and may come from internal genetic conflict, the most important for our species arising from differentially imprinted maternal and paternal genes. Selection also favors suppressing negative phenotypic traits. Finally, a positive form of self-deception may serve to orient the organism favorably toward the future. Self-deception can be analyzed in groups and is done so here with special attention to its costs. PMID- 10818625 TI - The evolution of moral dispositions in the human species. AB - Kohlberg's model of moral development is viewed from the perspective of evolutionary biology. Moral judgments defining Kohlberg's stages of moral development are seen as manifestations of structures evolved to uphold systems of cooperation. Game theory research on adaptive strategies of cooperation supports the conclusion that humans inherit dispositions to uphold the systems of cooperation implicit in the first three stages in Kohlberg's sequence, but not the systems of cooperation implicit in the highest stages. The empirical evidence on real-life morality is more consistent with a biological model of ontogenesis than is the model espoused by Kohlbergians. Although people occasionally make moral judgments in their everyday lives to reveal their solutions to moral dilemmas, as Kohlberg's model assumes, they more often make moral decisions that advance their adaptive interests. PMID- 10818626 TI - Genomic imprinting, sex-biased dispersal, and social behavior. AB - Some genes carry a record of the sex of the gene's carrier in the previous generation that influences the gene's expression in this generation. This additional information can result in intragenomic conflicts between an individual's maternally and paternally derived alleles over behaviors that affect relatives with whom the individual has different degrees of maternal and paternal relatedness. Asymmetries of relatedness can arise because of sex-biased dispersal. For example, if females remain in their natal group and males disperse, female members of a group will all be matrilineal relatives, but may have unrelated fathers. Sex-linked inheritance creates an evolutionary bias in favor of social groups that trace descent through the homogametic sex. This bias has a positive and negative aspect. The positive aspect is increased relatedness among siblings of the homogametic sex. The negative aspect is the lack of sex linked relatedness between parents and offspring of the heterogametic sex. PMID- 10818627 TI - Do extraterrestrials have sex (and intelligence)? AB - This thought experiment addresses the range of possible evolved psychologies likely to be associated with extraterrestrial (ET) intelligence. The analysis rests on: (1) a number of assumptions shared by the SETI project; (2) recent arguments concerning convergent evolution; and (3) current theories of how intelligence evolved in our own species. It concludes that, regardless of how and which cognitive abilities arise initially, extraterrestrially they can develop into intelligence only if an amplification process involving a form of predation and/or sexual selection occurs. Depending on the amplification process, ETs may be xenophobic; however, it is more probable that they will be ethnocentric. Their ideas of reciprocity and fairness are likely to at least overlap with our own. They will definitely be culture-bearing and probably have two sexes, both of which are intelligent. Regardless of the degree of physical similarity of ETs to ourselves, convergence makes it likely that we will at least find their evolved psychology similar enough to our own for comprehension. PMID- 10818628 TI - Freud: the first evolutionary psychologist? AB - An evolutionary perspective on attachment theory and psychoanalytic theory brings these two fields together in interesting ways. Application of the evolutionary principle of parent-offspring conflict to attachment theory suggests that attachment styles represent context-sensitive, evolved (adaptive) behaviors. In addition, an emphasis on offspring counter-strategies to adult reproductive strategies leads to consideration of attachment styles as overt manifestations of psychodynamic mediating processes, including the defense mechanisms of repression and reaction formation. PMID- 10818629 TI - Female reproductive strategies as social organizers. AB - When male philopatry is linked to patriarchy, it is often assumed that female reproductive strategies are secondary to male reproductive strategies. By comparing the social structures of chimpanzees (Pan troglodytes) and bonobos (Pan paniscus) with those of other non-human primates, I argue that female reproductive strategies can be viewed as primary principles of social organization, including the establishment of patriarchies through differential investment in offspring. Emphasis on the contributions of female reproductive strategies may lead to a different picture of the evolution of primate social organization. PMID- 10818631 TI - Mood as mechanism. PMID- 10818630 TI - Sex, sex differences, and social behavior. AB - Sex differences in social behavior are center stage in recent formulations of evolutionary psychology. Evolutionary psychology, with its emphasis on the long term consequences of early adaptations, offers itself as an alternative meta theory to mainstream social psychology, which emphasizes the importance of social structures in determining the existence and extent of social and cognitive sex differences. Using a range of examples, we argue that evolutionary psychology is open to criticism on several fronts: It does not (a) include a role for mediating and moderating variables or test predictions rigorously; (b) appreciate the importance of the difference between first- and second-order effects; (c) offer a truly interactionist theory; or (d) seriously consider the social implications of sex-based inequities. We also argue that social psychology has, in its turn, failed to appreciate the non-intuitive richness of some evolutionary hypotheses or that there is a role for evolutionary psychology in a genuinely interactionist theory This paper restates the need for that perspective, and suggests how it may be achieved. PMID- 10818632 TI - Resisting biology. The unpopularity of a gene's-eye view. AB - David Haig's intragenomic conflict theory concerning the possible evolutionary origins of genomic imprinting is discussed. It is suggested that a useful way in which one might represent Haig's theory to a popular audience would be by emphasizing a gene's eye viewpoint. In doing so, an author may help to dispel common misconceptions about evolution and natural selection, misconceptions which arise in part from an overemphasis on the individual, rather than the gene, when discussing natural selection. Although it is possible to view either the gene or the individual as a unit of natural selection, our own proclivity towards overemphasizing the role of individuals in evolution is seen as a common impediment to a more complete understanding of both natural selection, as well as the philosophical implications arising from a gene's eye view of evolutionary theory. PMID- 10818633 TI - On the evolution of misunderstandings about evolutionary psychology. AB - Some of the controversy surrounding evolutionary explanations of human behavior may be due to cognitive information-processing patterns that are themselves the result of evolutionary processes. Two such patterns are (1) the tendency to oversimplify information so as to reduce demand on cognitive resources and (2) our strong desire to generate predictability and stability from perceptions of the external world. For example, research on social stereotyping has found that people tend to focus automatically on simplified social-categorical information, to use such information when deciding how to behave, and to rely on such information even in the face of contradictory evidence. Similarly, an undying debate over nature vs. nurture is shaped by various data-reduction strategies that frequently oversimplify, and thus distort, the intent of the supporting arguments. This debate is also often marked by an assumption that either the nature or the nurture domain may be justifiably excluded at an explanatory level because one domain appears to operate in a sufficiently stable and predictable way for a particular argument. As a result, critiques in-veighed against evolutionary explanations of behavior often incorporate simplified--and erroneous -assumptions about either the mechanics of how evolution operates or the inevitable implications of evolution for understanding human behavior. The influences of these tendencies are applied to a discussion of the heritability of behavioral characteristics. It is suggested that the common view that Mendelian genetics can explain the heritability of complex behaviors, with a one-gene-one trait process, is misguided. Complex behaviors are undoubtedly a product of a more complex interaction between genes and environment, ensuring that both nature and nurture must be accommodated in a yet-to-be-developed post-Mendelian model of genetic influence. As a result, current public perceptions of evolutionary explanations of behavior are handicapped by the lack of clear articulation of the relationship between inherited genes and manifest behavior. PMID- 10818634 TI - Poiseuille Award Lecture. Viscometric, in vitro and in vivo blood viscosity relationships: how are they related? PMID- 10818635 TI - Adhesion of nonmetastatic and highly metastatic breast cancer cells to endothelial cells exposed to shear stress. AB - The mechanical stimulus of shear stress has to date been neglected when studying the adhesion of cancer cells to the endothelium. Confluent monolayers of endothelial cells were subjected to either 4 or 15 hours of arterial shear stress. Adhesion of nonmetastatic (MCF-7) and highly metastatic (MDA-MB-435) human breast cancer cells was then quantified using a detachment assay carried out inside the parallel plate flow chamber. Four hours of shear stress exposure had no effect on adhesion. However, 15 hours of shear stress exposure led to marked changes in the ability of the endothelial monolayer to bind human breast cancer cells. An increase in adhesive strength was observed for nonmetastatic MCF 7 cells, while a decrease in adhesive strength was observed for highly metastatic MDA-MB-435 cells. Hence, endothelial shear stress stimulation does influence the adhesion of cancer cells to the endothelium and can have different effects on the adhesion of cancer cells with different metastatic potentials. Furthermore, adhesion of nonmetastatic and highly metastatic human breast cancer cells may be controlled by two different endothelial cell adhesion molecules that are differentially regulated by shear stress. Immunohistochemistry confirmed that shear stress did in fact differentially regulate endothelial cell adhesion molecule expression. PMID- 10818636 TI - Computer modeling of fluid dynamics related to a myocardial bridge in a coronary artery. AB - Fluid mechanics associated with blood flows induced by the so-called myocardial bridge (MB) has been studied systematically using a computational fluid dynamic modeling of the Newtonian, incompressible, two-dimensional, unsteady flow in a channel with a time-dependently flushing in/out indentation. During each cycle, a train of vortex wave flow was observed downstream of the phasic stenosis and both upper and lower walls suffer severely from consistently high, oscillating wall shear stresses (WSS). Extensive studies were conducted on the influence of the Reynolds number, the geometry and the Strouhal number of the MB movement on the nature of the vortex flow and the time-dependent wall shear stress distribution. Special attention was drawn to the relationship between the vortex wave and the pressure distribution. It was found that the pressure gradient changed markedly during one cycle, which was apparently dominated by the dynamics of the indentation. A steep, adverse pressure gradient was observed when the indentation was flushing out, which corresponded to the existence of the most developing vortices. It implies the possibility that the MB in a coronary artery can produce an extremely low pressure region immediately downstream of the phasic stenosis, where elastic choking or collapse of the coronary artery might occur. PMID- 10818637 TI - Mechanical characteristics of the canine thoracic duct: what are the driving forces of the lymph flow? AB - This study is designed to better understand the mode of lymph transport, particularly through the extrinsic pumping by external compression of the lymph vessel. The pressure-diameter relationship of lymphatic segments isolated from the canine thoracic duct was examined using a laser optical micrometer measurement system. Results revealed that the thoracic duct displayed a high extensibility or compliance in the physiological pressure range, yet became progressively less so with increasing internal pressure. The calculated incremental circumferential modulus of the thoracic duct under physiological pressure (range of 2 to 6 cm H2O) showed values ranging from 1.2 x 10(4) to 3.61 x 10(5) dyn/cm2. At a pressure of 35 cm H2O, the modulus reached a limiting value of approximately 6.0 x 10(6) dyn/cm2. In the physiological pressure range, the relative wall thickness (h/R0) of the canine thoracic duct was approximately 3.5%, which was much lower than that reported for canine arterial segments and similar in value to that of the canine jugular vein. In conclusion, the pressure diameter curve of the canine thoracic duct was shown to resemble that of venous vessels. However, the circumferential elastic modulus of the thoracic duct wall was lower than the moduli of veins, proving that lymphatics are more compliant than veins. This suggests lymph flow in the thoracic duct may be better promoted by external compression of the lymphatic vessel. PMID- 10818638 TI - Swelling and flow properties of tubular poly(vinyl alcohol) gels. AB - Swelling and flow properties of tubular poly(vinyl alcohol) (PVA) hydrogels prepared with the cooling method were investigated using an inflation testing method. When the tubular hydrogel in liquid paraffin was inflated by using liquid paraffin as a pressure transmitting medium, namely in the case that the liquids inside and outside the gel are both liquid paraffin (P/P combination), the gel showed a slight volume change determined by Poisson's ratio of the gel. When the gel in water was inflated by liquid paraffin (P/W combination), the gel swelled to large extent compared with the case of P/P. The hydrogel in W/W combination, namely in the situation that the gel was immersed in water and also inflated by water, showed a very large volume change if the comparison was done at the same pressure. The origin of the volume change in P/P, P/W and W/W combinations is discussed. The volume change in P/P was governed by the Poisson ratio as a material constant (mu 0) of the PVA gels, and the gels swelled by the change in the application of pressure (or deformation) in P/W. The volume change in W/W was closely related to the flow of solvent in the gel. PMID- 10818639 TI - Dielectric approach to the investigation of erythrocyte aggregation: I. Experimental basis of the method. AB - A method based on dielectric properties of dispersed systems was developed to investigate red blood cell (RBC) aggregation in blood and RBC suspensions. Measurements of capacitance and resistance were made in a rectangular channel at low (0.2 MHz) and high (14 MHz) frequencies relative to the mid-point of the beta dispersion range. Compared to capacitance, minimal post-shearing changes of resistance were observed; capacitance changes at 0.2 MHz were two orders of magnitude larger than those at 14 MHz and hence subsequent measurements were carried out at the lower frequency. It is shown that post-shearing changes in the capacitance are affected by the recovery of RBC shape and relaxation processes at the electrode-suspension interface. However, the dominant factor contributing to time-dependent changes in the capacitance is the dynamic process of RBC aggregation. It is experimentally shown that the time record of the capacitance at 0.2 MHz quantitatively reflects the aggregation process in RBC-plasma suspensions with hematocrit up to 0.56 (v/v) and in suspensions of RBCs in artificial aggregating media. It is concluded that a dielectric approach to the study of RBC aggregation in whole blood offers great potential for basic studies and for diagnostic use. PMID- 10818640 TI - Cell electrophoresis studies relevant to red blood cell aggregation. PMID- 10818641 TI - Evidence against macromolecular "bridging" as the mechanism of red blood cell aggregation induced by nonionic polymers. PMID- 10818642 TI - Basic phenomena of red blood cell rouleaux formation. PMID- 10818643 TI - Characterization of erythrocyte aggregation with ultrasound. PMID- 10818644 TI - The effect of red blood cell aggregation on blood flow resistance. PMID- 10818645 TI - What do measurements of filterability of blood tell us about the effect of red cell aggregation on flow resistance in small vessels? PMID- 10818646 TI - Effects of red cell aggregation on the venous microcirculation. PMID- 10818647 TI - Effect of red blood cells and their aggregates on platelets and white cells in flowing blood. PMID- 10818648 TI - Serotonin syndrome: potentially fatal but difficult to recognize. PMID- 10818649 TI - The future for non-principal general practitioners: lost doctors--lost to whom? PMID- 10818650 TI - Antidepressants and the serotonin syndrome in general practice. AB - BACKGROUND: As a consequence of the greater use of agents affecting the serotonergic system, a syndrome of serotonin hyperstimulation has been recognized more frequently. The serotonin syndrome is characterized by a constellation of symptoms that include mental status changes, agitation, myoclonus, hyperreflexia, sweating, shivering, tremor, diarrhoea, lack of coordination, and fever. Deaths have been reported. AIM: To identify cases of the serotonin syndrome among patients prescribed a new antidepressant in general practice, and to determine doctors' awareness of the syndrome. METHOD: Patients who were dispensed nefazodone in England between 1996 and 1997 were identified using dispensed prescription data. Prescribing doctors were sent questionnaires as part of a post marketing surveillance study. Patients reported to have experienced two or more features of the serotonin syndrome were identified, and specific questionnaires were sent to their general practitioners. RESULTS: There was a 96.2% return rate of serotonin syndrome questionnaires. Nineteen cases met criteria for the syndrome (incidence = 0.4 cases per 1000 patient-months of treatment with nefazodone). Eight patients developed symptoms while taking nefazodone alone. Serotonergic symptoms were reported to a similar degree with five other antidepressants studied by the same method. In total, 85.4% of responding general practitioners were unaware of the serotonin syndrome. CONCLUSION: Improved awareness of the syndrome is needed within general practice. There is a need to distinguish the relatively minor serotonergic symptoms from those of a severe, life-threatening serotonin syndrome. PMID- 10818651 TI - Patients' reasons for not presenting emotional problems in general practice consultations. AB - BACKGROUND: Patients commonly do not mention emotional problems in consultations, and this is a factor in general practitioners' (GPs') difficulty in identifying psychological morbidity. AIM: To investigate patients' self-reported reasons for not disclosing psychological problems in consultations with GPs. METHOD: From nine general practices, a sample of patients with high General Health Questionnaire scores, who planned to present only somatic symptoms to the GP, were interviewed after their consultation with the GP. The interview covered their reasons for not mentioning emotional problems. A patient satisfaction questionnaire was administered. RESULTS: A total of 83 patients were interviewed. Sixty-four patients confirmed that they had not mentioned emotional problems in the consultation; 23 (36%) of these gave primarily realistic reasons for not presenting emotional problems (e.g. able to cope with distress), 29 (45%) gave reasons related to psychological embarrassment or hesitation to trouble the GP, and 12 (19%) were mainly deterred by the doctors' interview behaviours. The latter group had significantly lower satisfaction scores than patients in the other two groups. In addition, patients in all groups commonly reported perceptions of lack of time (48%) and that there is nothing doctors can do to help (39%) as barriers to mentioning emotional problems. CONCLUSION: An understanding of patients' reasons for not disclosing emotional problems can assist in identifying subgroups of patients with different management needs. PMID- 10818652 TI - Non-attendance at psychiatric outpatient clinics: communication and implications for primary care. AB - BACKGROUND: What should happen when an outpatient fails to attend a psychiatric clinic? Guidelines from the General Medical Services Committee suggest that general practitioners (GPs) have no further responsibility of care for a patient once a referral to a psychiatrist has been made. This raises questions about the formulation of effective management plans for those patients with whom psychiatric services find it difficult to engage due to non-compliance with assessment and follow-up. AIMS: To investigate communication between GPs, patients, and psychiatrists at referral and following attendance or non attendance at outpatient appointments. METHOD: A prospective study of a random sample of attenders and non-attenders at psychiatric outpatient clinics. Patients and GPs were interviewed to obtain data about the referral process. GPs' views on communication from psychiatrists and the appropriate course of action following clinic non-attendance were investigated. The quality of referral and clinic letters for attenders and non-attenders was compared. RESULTS: There was no difference in the quality of referral letter for attenders and non-attenders. Psychiatrists were less likely to write to GPs about follow-up patients' appointments than new patients' appointments; communication was least likely when a follow-up patient missed their appointment. GPs considered follow-up non attenders were more likely to need a further appointment than new patient non attenders, but did not identify a role for themselves in engaging with follow-up non-attenders. CONCLUSION: Communication between GPs and psychiatrists about new patients seems adequate. However, there are important deficits in communication from psychiatrists to GPs about follow-up patients, especially non-attenders who are often more severely ill and difficult to engage. An effective response for this group is likely to need cooperative health and social service action rather than rigid guidelines concerning clinical responsibility. PMID- 10818653 TI - Morbidity, deprivation, and antidepressant prescribing in general practice. AB - BACKGROUND: Although the link between depression, unemployment, and measures of deprivation and morbidity has been previously documented, the relationship between general practice prescribing of antidepressants, morbidity, and the social demography of general practice populations is poorly understood. AIM: To consider whether morbidity and the social demography of general practice populations influence the prescribing costs of individual practices. METHOD: Data were analysed, using a forward stepwise regression procedure, of all 78 practices served by the Cornwall and Isles of Scilly Health Authority. Data on prescribing for antidepressants were provided by the Prescription Pricing Authority for the period from July to December 1995 and converted into defined daily doses (DDDs) to standardize for the variation in prescribing practice between general practitioners. RESULTS: A significant positive correlation exists between the rates of prescribing DDDs of antidepressants by general practices and the prevalence of permanent sickness in the areas in which these practices serve. CONCLUSION: Demonstrating an association between morbidity and prescribing rates for depression may prove helpful in setting prescribing budgets. PMID- 10818654 TI - Factors explaining the use of psychiatric services by general practices. AB - BACKGROUND: Referral rates from general practitioners to hospital services vary up to 25-fold, and several studies have sought reasons for this apparent inconsistency in clinical practice. However, few studies have concentrated on, or indeed included, psychiatric patients or psychiatric referral rates. AIM: To determine the effect of population, general practice, and mental health service factors on use of specialist mental health services by general practices. METHOD: Cross-sectional data from computerized records used in managing clinical care on all patients aged 16 to 64 years who had been in contact with any mental health service staff over a two-year period. Twenty-three practices in Huntingdon Health District were studied, with a list population of 87,643 patients aged 16 to 64 years, served by one inpatient ward and three community mental health teams. The main outcome measures were the relation between age-standardized utilization ratio and markers of morbidity, deprivation, community mental health provision, and practice prescribing. RESULTS: Variation between practices in the use of mental health services was relatively limited, especially compared with the use of other secondary medical and surgical services. Three factors together explained 60.8% of the variance in use between practices: a census-based index of long-term limiting illness in females registered with the practice, use of one of the three community mental health teams, and average quarterly defined daily doses of hypnotics prescribed per practice population. Relatively high prescribing of hypnotics was associated with lower service use. CONCLUSION: Population morbidity and factors in the mental health service explain a substantial part of the variation in the use of mental health services between practices. Further work is needed to replicate these findings and explore why team factors and prescribing patterns influence utilization ratios. This study underlines the importance of examining population, practice, and specialist service factors in explaining variation in the use of secondary care by general practices. PMID- 10818655 TI - Newer antidepressants: a comparison of tolerability in general practice. AB - BACKGROUND: An increasing number of antidepressants have been released on the United Kingdom market in recent years, and these are being prescribed more frequently in general practice. Clinical trials suggest that such agents have similar efficacy and the choice of drug is probably based on tolerability, toxicity in overdose, and cost. AIM: To compare the tolerability and safety profile of six, newly marketed antidepressants used in general practice. METHOD: Studies have been conducted for six antidepressants: fluoxetine, sertraline, paroxetine, moclobemide, venlafaxine, and nefazodone, using the technique of prescription-event monitoring. Patients were identified using incident dispensed prescription data. Questionnaires were sent to patients' general practitioners six months after the date of first prescription. Questionnaires asked for date of birth, sex, indication for prescribing each drug, and all events entered in the patients' records after the date of first prescription. RESULTS: Each cohort exceeded 10,000 patients. Nausea/vomiting was the most frequently reported event for all drugs. The difference in incidence rates for drowsiness/sedation, male sexual dysfunction, and hypertension is shown. Mortality data are also reported. CONCLUSION: Frequently reported events were similar for all six drugs but there were clinically and statistically significant differences for less frequently reported events. The adjusted mortality rate was identical between the six drugs. This study provides valuable comparative data for six, widely used antidepressants in general practice. PMID- 10818656 TI - Urinary symptoms and incontinence in women: relationships between occurrence, age, and perceived impact. AB - BACKGROUND: The prevalence of urinary symptoms that impact on quality-of-life will be important in determining resource allocation in primary care groups. AIM: To determine the prevalence of urinary symptoms and their perceived impact in a community population of women. METHOD: A postal survey using a validated self completed questionnaire among all women aged over 18 years and registered with one general practice in a major British city. The prevalence rates and perceived impact of a wide range of urinary symptoms and their relationship with age was determined. Data were analysed using the chi-squared test and the chi-squared test for trend. Spearman's rank correlation was used to assess the relationship between symptom severity and perceived impact. RESULTS: The number of completed questionnaires returned was 2075, giving an 80% response rate. Of these, the number of women who reported some degree of incontinence in the previous month was 1414 (69%), although only 578 (30%) indicated that it had social or hygienic impact. Other lower urinary tract symptoms reported included nocturia (19%), poor stream (19%), urgency (61%), and dysuria (23%). The most troublesome symptoms were incontinence for no obvious reason, nocturnal incontinence, and nocturia, with 73%, 69%, and 63% of sufferers, respectively, finding these symptoms troublesome. CONCLUSIONS: Incontinence and other urinary symptoms are more common than previously thought. These symptoms are not always perceived as bothersome or as having a social or hygienic impact, and therefore many women who report urinary leakage do not require treatment. Nocturnal symptoms in women are commoner than might have been supposed and are extremely troublesome to sufferers. PMID- 10818657 TI - Vaccine, yes; injection, no: maternal responses to the introduction of Haemophilus influenzae type b (Hib) vaccine. AB - We used in-depth interviews with 23 mothers of babies aged 1-2 years to explore their response to the addition of Haemophilus influenzae type b (Hib) vaccine to the primary schedule. Acceptability of the vaccine was principally attributable to maternal perceptions as to the severity of meningitis, with a tendency to overestimate the efficacy of the vaccine. Advice from health professionals and the behaviour of peers contributed to its acceptance. Barriers to the uptake of the vaccine included suspicion regarding the newness of the vaccine, a fear of vaccine overload in such young babies, and the distress of two separate injections. There may be a limit to the number of antigens, and particularly the number of injections, acceptable to mothers. PMID- 10818658 TI - Locality commissioning: how much influence have general practitioners really had? AB - This paper investigates the various models of locality commissioning in relation to the participation of general practitioners (GPs), and explores the perceived successes of locality commissioning in the 15 health boards in Scotland and 13 health authorities in the Northern and Yorkshire Region of England. A postal questionnaire was sent to 190 individuals involved in commissioning, and semi structured interviews with GPs (n = 31) and health authority managers (n = 41) were undertaken in each of the 28 health authorities. Seventy-five per cent of the health authorities had introduced some form of locality commissioning. Five types of locality commissioning organization were identified on the basis of the level of GP influence over decisions. All GP responders identified benefits resulting from their involvement in the process but only 27% of health authority responders did so. Most benefits related to improved professional relationships, not to service changes. On the whole, locality commissioning does not appear to have resulted in major changes to contracts or services. PMID- 10818659 TI - 'Bin bag' study: a survey of the research requests received by general practitioners and the primary health care team. AB - General practitioners receive a large and increasing number of unsolicited requests to participate in research. This study describes the volume and nature of research requests received by 18 primary care teams in a three-month period. On average, each practice receives 16 to 24 research requests each year. The most frequent request is to complete a questionnaire (32%). Only one-fifth of studies originate from academic or service general practice. Remuneration for participating in a study was only offered for 15% of studies. Although general practice teams feel swamped by research requests, this sensation may be exaggerated by invitations to participate in non-scientific surveys as well as true research projects. Practice teams would welcome help in distinguishing quality research proposals from the remainder. PMID- 10818660 TI - Acceptability of screening young children for anaemia. AB - Although anaemia is common among young children and may be detrimental to health and development, few blood tests are done in this age group. We found that thumb prick blood tests were not stressful to most young children and, despite the high mobility of the population, achieved an 81% uptake of screening for anaemia (273 out of 335 eligible children). PMID- 10818661 TI - They call it stormy Monday--reasons for referral from primary to secondary care according to the days of the week. AB - Previous studies of specialist care have shown that the onset of acute myocardial infarction occurs most frequently on Mondays; however, the septadian pattern in the reasons for referrals from primary care to secondary care has not been examined previously. We examined daily variations in rates of referral from primary to secondary care in central and northern Finland, using International Classification of Primary Care codes, during two weeks (30 November to 6 December 1992, and 28 November to 4 December 1994) before and after the introduction of a reform in the system of financing health care. Monday peaks were observed in the occurrence of angina pectoris, fractures of the tibia/fibula, lumbar disc lesions, and infections of the eye. Tuesday peaks were found in the occurrence of headache. Two peaks (on Tuesday and Thursday) were observed for patients with suspected appendicitis. Many problems of primary health care leading to referral to secondary care are not random events but occur in a weekly pattern. Further investigation is needed within these subgroups of patients to identify the causes of this variation. PMID- 10818663 TI - Conducting randomized trials in general practice: methodological and practical issues. AB - The evaluation of the outcome of health services technologies is a requirement for their efficient provision in clinical practice. The most reliable evidence for treatment efficacy comes from randomized trials. Randomized trials in general practice pose particular methodological and practical difficulties. In this paper, we discuss how best to plan and manage a clinical trial in this setting. We base our discussion on our experience of conducting randomized trials to evaluate the effectiveness of brief psychotherapy in general practice. PMID- 10818662 TI - Should sexual partners of women with bacterial vaginosis receive treatment? AB - Bacterial vaginosis is the most prevalent infectious cause of vaginitis. It is associated with significant morbidity, particularly in pregnant women and following gynaecological operations. Cure is difficult. There is some controversy over whether treating sexual partners of affected women can improve cure rates. This paper provides a critical appraisal of the evidence for simultaneously treating the male partner of women affected by bacterial vaginosis. Unfortunately, no evidence was found supporting the treatment of partners of women affected by bacterial vaginosis. PMID- 10818664 TI - SPREAD initiative. The Research Team. PMID- 10818665 TI - Screening for cervical chlamydial infection in general practice. PMID- 10818666 TI - Locum doctors in general practice: motivation and experiences. PMID- 10818667 TI - The need for an eBJGP. PMID- 10818669 TI - PACT quarterly returns. PMID- 10818668 TI - Antibiotic prescribing patterns. PMID- 10818670 TI - Influenza virus sialidase: a target for drug discovery. PMID- 10818671 TI - Fibrinogen receptor antagonists: design and clinical applications. PMID- 10818672 TI - Combinatorial chemistry as a tool for drug discovery. AB - The question 'will combinatorial chemistry deliver real medicines' has been posed [96]. First it is important to realise that the chemical part of the drug discovery process cannot stand alone; the integration of synthesis and biological assays is fundamental to the combinatorial approach. The results presented in Tables 3.1 to 3.8 suggest that so far smaller directed combinatorial libraries have obtained equivalent results to those obtained previously from traditional medicinal chemistry analogue programs. Unfortunately, because of the long time it takes to develop pharmaceutical drugs there are no examples yet of marketed drugs discovered by combinatorial methods. There are interesting examples where active leads have been discovered from the screening of the same library against multiple targets (e.g. libraries 13, 39, 43, 66, 71 and 76). It is now possible to handle much larger libraries of non-oligomeric structures and the chemistry required for such applications is becoming available. Whether combinatorial approaches can also be adapted to deal with all the other requirements of a successful pharmaceutical (lack of toxicity, bioavailability etc.) is open to question but there are already examples such as cassette dosing [235-237]. However we can still be optimistic about the possibility of larger libraries producing avenues of investigation for the medicinal chemist to develop into real drugs. Combinatorial chemistry is an important tool for the medicinal chemist. PMID- 10818673 TI - New directions in anxiolytic drug research. AB - Agents to treat anxiety have gained in acceptance and importance in the fast pace of life in the second half of this century. The discovery and refinement of the benzodiazepines represented a quantum leap in therapy from early compounds which were essentially sedatives. With the advent of molecular biology, an understanding of the basic mechanism by which the benzodiazepines exert their effects was revealed through the discovery and isolation of the GABAA receptor and its benzodiazepine binding site. This, in turn, has enabled benzodiazepines to be classified into a broad spectrum of pharmacological types ranging from agonist to inverse agonist, thus allowing fine tuning with respect to side effects. Consequently, newer, more promising agents have emerged which bind at the GABAA BZD site and have reduced side-effects. An example of this is RWJ-51204 (92), a member of a novel structural type which is superior to several marketed benzodiazepines in animals in terms of efficacy and side-effects. The cost conscious environment of managed health care presents continuing challenges to the discovery and development of safe, highly efficacious, and cost-effective anxiolytic agents. PMID- 10818674 TI - Selective inhibitors of cyclooxygenase-2 (COX-2). PMID- 10818675 TI - Molecularly imprinted polymers--preparation, biomedical applications and technical challenges. PMID- 10818676 TI - New paradigms for the treatment of cancer: the role of anti-angiogenesis agents. AB - Angiogenesis, the sprouting of new blood vessels, plays a role in diverse disease states including cancer, diabetic retinopathy, age-related macular degeneration, rheumatoid arthritis, psoriasis, atherosclerosis, and restenosis. With regard to cancer, the clinical association of tumor vascularity with tumor aggressiveness has been clearly demonstrated in numerous tumor types. The observation of increased microvessel density in tumors not only serves as an independent prognostic indicator, but also suggests that anti-angiogenic therapy may be an important component of treatment regimens for cancer patients. The complexity of the angiogenic process, which involves both positive and negative regulators, provides a number of targets for therapy. Many positive regulators, including growth factor receptors, matrix metalloproteinases, and integrins, have been correlated with increased vascularity of tumors and poor prognosis for patient survival. Thus, these serve as ideal targets for anti-angiogenesis therapy. Many inhibitors of these targets are currently undergoing clinical evaluation as potential anti-cancer agents. In this article, we discuss the role of positive regulators in angiogenesis and tumor growth and describe the anti-angiogenic agents under development. PMID- 10818677 TI - The hepatocyte growth factor/Met pathway in development, tumorigenesis, and B cell differentiation. AB - This article summarizes the structure, signal transduction and physiologic functions of the HGF/Met pathway, as well as its role in tumor growth, invasion, and metastasis. Moreover, it highlights recent studies indicating a role for the HGF/Met pathway in antigen-specific B-cell development and B-cell neoplasia. PMID- 10818678 TI - Clinical targets for anti-metastasis therapy. AB - Metastasis is responsible for most cancer deaths. Therapeutic strategies to prevent development of metastases thus have potential to impact on cancer mortality. Development of these therapies requires a better understanding of the biology and molecular events of the metastatic process. Metastasis is usually defined, clinically and experimentally, by evidence of the endpoint of the process, that is, the presence of metastatic tumors. Endpoint assays are suitable for determining if a therapeutic approach is effective, but can provide little information on how a treatment works in vivo and what steps in metastasis are affected. We describe here two methodological advances in the study of metastasis as a process: in vivo videomicroscopy, which permits direct observation of steps in metastasis, and a "cell accounting" technique that permits quantification of the fate of cells over time. These procedures have provided new and unexpected insights into the biology of the metastatic process. Based on these insights, we consider which steps in the metastatic process are biologically and clinically most appropriate as therapeutic targets for development of anti-metastasis therapies. We conclude that the most promising stage of the metastasis process for therapeutic targeting is the growth phase, after cancer cells have arrested in the microcirculation in secondary sites and have completed extravasation. Earlier phases in the process are either biologically inappropriate or clinically inaccessible, except in specific cases (e.g., chemoprevention strategies). The role of "seed" and "soil" in determining organ-specific metastasis is also discussed. The metastatic growth phase fortunately is a clinically broad target, and any treatment that limits growth of metastases prior to their causing irreversible harm to the patient has the potential to be clinically useful. A variety of therapeutic approaches to target this phase are under active development, including inhibition of angiogenesis or signal transduction pathways needed to support the growth of metastatic cells. PMID- 10818679 TI - Animal models of melanoma: recent advances and future prospects. PMID- 10818680 TI - The indispensable role of microenvironment in the natural history of low-grade B cell neoplasms. AB - Follicular lymphoma (FL) and B-cell chronic lymphocytic leukemia (B-CLL) are paradigmatic examples of lymphoid malignancies in which the relevant biological mechanisms are alterations in the control of apoptosis rather than an exaggerated proliferation. This explains why low-grade B-cell neoplasms still fail to be cured with current approaches. It is becoming increasingly clear that the defective apoptosis of FL and B-CLL has to be ascribed not only to intrinsic defects of the neoplastic cells, but also to extrinsic factors that influence their behavior. Malignant B cells retain the capacity to respond to microenvironmental signals, but have devised a monothematic responsiveness. They have a specific sensitivity to anti-apoptotic signals that favor their survival, whereas they seem to have become insensitive to pro-apoptotic signals. Bystander, nontumoral cells play a fundamental (though not sufficient) role both in the onset and in the progression of these diseases. The survival of leukemic cells appears to be dependent on direct cell-cell contacts. The localization of malignant B cells in bone marrow or neoplastic follicles is not a passive adhesion phenomenon but a crucial step for their survival. Bidirectional malignant lymphocyte-nontumoral cell interactions may lead to the amplification of a microenvironment able to inhibit the apoptosis of neoplastic B cells. The pressure of antigenic selection and the role of the tumor necrosis factor receptor family through the functional survival signal provided by CD40 together with the crippled death signal exerted by CD95 are new prominent characters on the stage. PMID- 10818681 TI - Epstein-Barr virus latency: LMP2, a regulator or means for Epstein-Barr virus persistence? AB - Like other herpesviruses, Epstein-Barr virus (EBV) persists in its host through an ability to establish a latent infection that periodically reactivates, producing infectious virus that infects naive hosts. Disease syndromes in humans caused by EBV reflect the cell types that EBV infects, being primarily of lymphoid or epithelial origin. The most notable lymphoid disease, infectious mononucleosis, is a self-limiting lymphoproliferative disease that occurs in normal adolescents on primary infection. Children are normally able to resolve primary EBV infection with few or no symptoms. By the age of 25 most individuals are EBV seropositive. EBV is associated with a variety of hematopoietic cancers such as African Burkitt's lymphoma, Hodgkin's, and adult T-cell leukemia. EBV associated lymphoproliferative disease occurs in individuals with congenital or acquired cellular immune deficiencies. The two notable epithelial diseases associated with EBV infection are nasopharyngeal cancer, a malignancy endemic to southern China, and oral hairy leukoplakia, an epithelial hyperplasia of the lingual squamous epithelium in AIDS patients. Latent membrane protein 2 (LMP2) is expressed both in normal EBV latency and EBV-associated pathologies. LMP2 may regulate reactivation from latency by interfering with normal B-cell signal transduction processes and in doing so may also provide a survival signal that could be important for viral persistence. Current knowledge about the function of LMP2 is described, defining a new class of regulators of herpesvirus latency. PMID- 10818682 TI - Biochemistry and pathological importance of mucin-associated antigens in gastrointestinal neoplasia. PMID- 10818683 TI - Studies on polyomavirus persistence and polyomavirus-induced tumor development in relation to the immune system. PMID- 10818684 TI - Improved nonradioactive RT-PCR method for relative quantification of mRNA. PMID- 10818685 TI - Epitope swapping to distinguish transgenic from endogenous fibroblast growth factor receptor type 1. PMID- 10818686 TI - Characterization of flanking sequences using long inverse PCR. PMID- 10818687 TI - Using the SMART cDNA system to map the transcription initiation site. PMID- 10818688 TI - Elevated temperature greatly improves transformation of fresh and frozen competent cells in yeast. PMID- 10818689 TI - Microdissection by exclusion and DNA extraction for multiple PCR analyses from archival tissue sections. PMID- 10818690 TI - Direct imaging of northern blots on an optical scanner using ethidium bromide. PMID- 10818691 TI - Accelerated titering of adenoviruses. PMID- 10818692 TI - BLAST ... what next? PMID- 10818693 TI - The 39 steps: a cautionary tale of quantitative 3-D fluorescence microscopy. PMID- 10818694 TI - Analysis of recombinant protein expression by MALDI-TOF mass spectrometry of bacterial colonies. AB - E. coli expressing soluble recombinant HIV antigens were analyzed directly by MALDI-TOF mass spectrometry (MS) from bacterial colonies picked from agar plates. An HIV envelope (ENV) antigen construct, penvA, was expressed in E. coli by transformation of the plasmid pPL/penvA-M. The plasmid was co-transformed into E. coli DH5 alpha cells with an equal quantity of the plasmid pKRR826, the parent vector without the penvA insert, and plated at medium density on L-agar plus ampicillin plates. A total of 24 colonies from four agar plates (six colonies per plate) were picked and transferred into 50% acetonitrile--0.1% trifluoroacetic acid aliquots for analysis by MALDI-TOF MS. The MS analysis detected 10 of 24 colonies expressing the recombinant protein; one colony expressed a mutant penvA protein; eleven of 24 colonies showed ions only from E. coli; and two of 24 colonies showed no detectable proteins. When E. coli transformed only with plasmid pPL/penvA-M were examined, all (10 of 10) colonies showed the penv insert by the MALDI-TOF MS method. The method is fast (less than 1.5 h for 24 colonies) and allows identification of colonies expressing intact or mutant proteins directly from culture plates without sample purification. PMID- 10818695 TI - Nonspecific, nested suppression PCR method for isolation of unknown flanking DNA. AB - We report the development of a simple, sensitive and robust two-step PCR method for the isolation of unknown sequences flanking characterized regions of genomic DNA or cDNA. The method requires 100 bp or less of a known sequence upstream of an oligonucleotide primer binding site. A first round of suppression PCR is conducted at low stringency with a polymerase lacking exonuclease activity to generate a mixture of products including fragments of the desired flanking sequence that are often greater than 1 kb in length. The desired fragments are then amplified from the mixture in a second round of suppression PCR using an extended oligonucleotide in combination with a polymerase exhibiting exonuclease activity. These fragments are subsequently identified by hybridization with the 100 bp of known sequence or simply by cloning and sequencing. The method is widely applicable and allows isolation of novel cDNA from very low abundance transcripts. PMID- 10818696 TI - Microcapillary reactors using solid-phase DNA sequencing for direct sample introduction into slab gels. AB - Solid-phase micro-reactors have been prepared in glass capillaries for DNA sequencing applications using slab gel electrophoresis, which consisted of a fused silica capillary (i.d. = 100 microns; o.d. = 365 microns; length = 15 cm; volume = 1.2 microL) that contained a covalently bound biotin molecule. With the addition of streptavidin to the capillary, an anchoring site was produced for the tethering of biotinylated DNA sequencing templates to the wall of the capillary. Using a four-lane, single dye primer chemistry sequencing strategy, the individual tracts were prepared in the capillaries using cycle sequencing (20 thermal cycles) on a PCR-generated lambda-bacteriophage template (about 1000 bp). The dye label in this case was a fluorescent tag that displayed emission properties in the near-IR and could be processed on an automated sequencer. The read length was found to be 589 bases, which was determined primarily by the fractionating power of the gel. It was also found that the tethering system was very stable to typical cycle sequencing conditions, with the amount of tethered DNA lost amounting to 40% after 120 thermal cycles. The ability to use dye terminator chemistry was also investigated by using a near-IR dye-labeled terminator (ddGTP). It was found that the quality of the ladder that was generated was comparable to that obtained in a conventional sample preparation format. However, ethanol precipitation was required before gel loading to remove excess terminator. PMID- 10818697 TI - Modified inter-simple sequence repeat PCR protocol for use in conjunction with the LI-COR Gene ImagIR2 DNA analyzer. AB - Inter-simple sequence repeat (inter-SSR) PCR was assessed for use in variety testing of chrysanthemum. This method was modified to allow detailed analysis of DNA profiles on a LI-COR Gene ImagIR2 DNA analyzer system. Protocols for unlabeled PCR were unsuccessful in producing labeled products when using infrared (IR) dye-labeled primers. Various modifications to the known protocols were investigated: (i) different ratios of labeled to unlabeled primer; (ii) various annealing temperatures; (iii) the use of an IR genotyping kit; (iv) end labeling; and (v) direct incorporation and cycle labeling. Successful amplification using labeled primers only occurred when two consecutive reactions were performed. The first PCR was performed using standard protocols for unlabeled reactions. The second PCR used a dilution of these reaction products as a template and 50% IR labeled and unlabeled primer. The complete procedure leading to a high-resolution analysis of inter-SSR PCR products on a LI-COR system is reported for the first time. This system allows high-throughput fingerprinting with the potential for applications on a commercial scale. PMID- 10818698 TI - Evaluation of cell membrane electropermeabilization by means of a nonpermeant cytotoxic agent. AB - For the evaluation of cell membrane electropermeabilization, cells are usually exposed to electric pulses in the presence of propidium iodide, a fluorescent dye activated by binding to cellular DNA. The fraction of permeabilized cells is then determined using a flow cytometer. This widely established method has several drawbacks: (i) an arbitrary choice of minimum fluorescence intensity for characterization of permeabilized cells; (ii) the inability to detect cells disintegrated because of intense electropermeabilization; and (iii) false detection of cellular ghosts devoid of fluorescence because of leakage of DNA caused by electropermeabilization. Here, we present a simple and inexpensive method that eliminates these drawbacks. The method is based on the use of a cytotoxic agent that cannot permeate through an intact plasma membrane and thus leads to selective death of the electropermeabilized cells. The amount of nonpermeabilized cells is then determined by a suitable viability test. Bleomycin at a 5-nM concentration causes no statistically significant effect on cell survival in the absence of electric pulses, yet this concentration is sufficient for lethal toxicity in electropermeabilized cells. The amount of cells surviving the exposure relative to the control gives a reliable value of the fraction of nonpermeabilized cells. PMID- 10818699 TI - Rapid determination of membrane transport parameters in adherent cells. AB - Reported here is a new method that permits rapid (approximately 5 s) determinations of membrane transport phenomena in cells grown in monolayers at the base of 17-mm glass scintillation vials. The method is convenient, cost effective and requires no special apparatus. Initial uptake rates, steady-state and free substrate levels are demonstrated in ZR-75-1 breast cancer and Chinese hamster ovary cell lines using methotrexate, a model agent transported by the reduced folate carrier. The technique should be applicable to the study of the transport properties in a broad range of substrates and cells in monolayer culture. PMID- 10818700 TI - Detection of Glu-Glu-tagged proteins in mammalian cell culture media by dot immunoblotting. AB - A dot immunoblotting technique has been developed to estimate the relative expression levels of tagged recombinant human proteins in mammalian cell culture media. Variations in sample denaturation, blocking agents and membrane composition and treatment were used to optimize the signal-to-noise ratio of the defined procedure. The method is rapid, with sensitivity extending to the low nanomolar range for a number of recombinant proteins. This technique should have general utility for antibody-based measurements of other tagged and non-tagged proteins in cell culture media or in biological fluids. PMID- 10818701 TI - Extraction of nuclear proteins with increased DNA binding activity. AB - We have developed a method for the rapid extraction of nuclear proteins from cultured cells. The ammonium sulfate method described here extracts larger quantities of proteins that retain DNA binding activity than the modified Dignam method and another popular method used for the extraction of transcription factors. The ammonium sulfate method is rapid and can be used to process a large number of samples for gel shift analysis. PMID- 10818702 TI - Making blind robots see: the synergy between fluorescent dyes and imaging devices in automated proteomics. AB - Proteomics investigations endeavor to provide a global understanding of gene product synthesis rate, degradation rate, functional competence, posttranslational modification, subcellular distribution and physical interactions with other cell components. Protein expression encompasses an enormous dynamic range. Since rare proteins cannot be amplified by any type of PCR method, sensitive detection is critical to proteome projects. Fluorescence methods deliver streamlined detection protocols, superior detection sensitivity, broad linear dynamic range and excellent compatibility with modern microchemical identification methods such as mass spectrometry. Two general approaches to fluorescence detection of proteins are currently practiced: the covalent derivatization of proteins with fluorophores or noncovalent interaction of fluorophores either via the SDS micelle or through direct electrostatic interaction with proteins. One approach for quantifying fluorescence is to use a photomultiplier tube detector combined with a laser light scanner. In addition, fluorescence imaging is performed using a charge-coupled device camera combined with a UV light or xenon arc source. Fluorescent dyes with bimodal excitation spectra may be broadly implemented on a wide range of analytical imaging devices, permitting their widespread application to proteomics studies and incorporation into semiautomated analysis environments. PMID- 10818703 TI - Improved method for detecting differentially expressed genes using cDNA indexing. AB - In cDNA indexing, differentially expressed genes are identified by the display of specific, corresponding subsets of cDNA. Subdivision of the cDNA population is achieved by the sequence-specific ligation of adapters to the overhangs created by class IIS restriction enzymes. However, inadequate specificity of ligation leads to redundancy between different adapter subsets. We evaluate the incidence of mismatches between adapters and class IIS restriction fragments during ligation and describe a modified set of conditions that improves ligation specificity. The improved protocol reduces redundancy between amplified cDNA subsets, which leads to a lower number of bands per lane of the differential display gel, and therefore simplifies analysis. We confirm the validity of this revised protocol by identifying five differentially expressed genes in mouse duodenum and ileum. PMID- 10818704 TI - Use of green fluorescent protein/Flp recombinase fusion protein and flow cytometric sorting to enrich for cells undergoing Flp-mediated recombination. AB - Flp recombinase has been used extensively for in vivo manipulation of eukaryotic DNA at specific sequences designated as FRT sites. We developed a method to use Flp-mediated recombination without the need for drug resistance or metabolic selection of cells in which recombination has occurred. We generated expression plasmids directing expression of fusion proteins consisting of Flp recombinase and green fluorescent protein (GFP) coding sequences. When the plasmids were introduced into K562 cells containing Flp recombinase substrates and transfected cells were selected for by flow cytometric sorting, GFP-positive cells were enriched 5- to 30-fold for Flp-mediated recombination events compared with unsorted cells. These studies demonstrate the usefulness of GFP/Flp recombinase fusion proteins to manipulate chromosomal DNA in vivo without requiring drug resistance or metabolic marker genes. PMID- 10818705 TI - Solubilization and display of G protein-coupled receptors on beads for real-time fluorescence and flow cytometric analysis. AB - G protein-coupled receptors (GPCR) and cellular signaling elements are prime targets for drug discovery. Sensitive real-time methods that expand the analytical capabilities for these elements can play significant roles in basic research and drug discovery. Here, we describe novel approaches for the real-time fluorescence analysis of GPCRs. Using the G protein-coupled N-formyl peptide receptor (FPR) as a model system in concert with a fluorescent ligand, we showed the quantitative solubilization of his-tagged FPRs in 1% dodecyl maltoside. Solubilized receptors reconstitute in dodecyl maltoside with a mixture of bovine brain Gi/Go showing an apparent Kd of 100 nM. Solubilized receptors were also bound to Ni(2+)-silica particles and were detected in a flow cytometer by the binding of fluorescent ligand. The efficiency of receptor uptake by the particles was in excess of 80% with an apparent affinity for the bead in the nM range. The receptors had largely homogeneous dissociation characteristics, an appropriate Kd for the ligand in the low nM range and a high site number, with several million receptor molecules per particle. However, the G protein reconstitution was not detected on the beads, apparently for steric reasons. These approaches for displaying receptors could prove useful in drug discovery and in the analysis of the molecular assemblies in signal transduction. PMID- 10818706 TI - Validation of fluorescent SSCP analysis for sensitive detection of p53 mutations. AB - We have developed a fluorescence-based single strand conformation polymorphism (SSCP) method that offers fast and sensitive screening for mutations in exons 5-8 of the human p53 gene. The method uses an ABI 377 DNA sequencer for unique color detection of each strand, plus accurate alignment of lanes for better detection of mobility shifts. To validate the method, 21 cell lines with reported mutations in p53 exons 5-8 were analyzed by SSCP using various gel conditions. The sensitivity for mutation detection was 95% for all cell lines studied, and no false positives were seen in 10 normal DNA samples for all four exons. Experiments mixing known amounts of tumor and normal DNA showed that mutations were detected even when tumor DNA was mixed with 80% normal DNA. Fluorescent SSCP analysis using the ABI sequencer is a useful tool in cancer research, where screening large numbers of samples for p53 mutations is desired. PMID- 10818707 TI - Development of a high-throughput process for detection and screening of genetic polymorphisms. AB - A method is described that uses the ABI PRISM 310 genetic analyzer in conjunction with custom-designed software to identify and classify RAPD products. This methodology will also work well with AFLPs and microsatellite analyses. The methodology uses the ABI PRISM 310's high-throughput (> 500 samples per week) capabilities and in-lane molecular weight standards to efficiently separate and size DNA products. Peak detection, locus classification and export of the data in a form accessible by several genetic analysis programs were accomplished through a custom-written software program (Peaks). Various criteria used by the program to identify and classify loci are described, and their effect on population analyses is examined. Criteria providing an effective, robust determination of population structure are presented. PMID- 10818708 TI - Focal extraction of surface-bound DNA from a microchip using photo-thermal denaturation. AB - High-throughput, selective extraction of a particular DNA fragment from a mixture of DNA before PCR amplification is becoming increasingly important in the DNA analysis field. Although the latest microchip technology has enabled real-time DNA expression analysis using hybridization between surface-bound probe DNA and sample DNA, the potential of this technology in purification of a small amount of DNA has not been demonstrated. We report here a method for area-selective release and collection of specific DNA, in which an IR laser beam is focused onto surface bound sample DNA at the target-spotted area to denature hybridized DNA. First, sample DNA labeled with a fluorescent dye was hybridized to a probe DNA immobilized on a chromium-coated chip. A 1053-nm IR laser beam with an intensity of 10-100 mW was then focused on the target area with a spatial resolution of 10 microns, causing the release of the fluorophore-labeled sample DNA as a result of photo-thermal denaturation. Confirmation of the amount of eluted DNA by PCR amplification after collection indicated that more than 10(-20) mol DNA/micron 2 area was eluted from the microchip, representing more than 70% of the chip-bound sample DNA. These results indicate that this method can be applied to the highly sensitive purification of DNA in microchip technology. PMID- 10818709 TI - Method for cloning restriction fragments containing the termini of BAC inserts. AB - We have developed a method to isolate the termini of BAC clones. The method is based on the two unique NotI sites located approximately 300 bp on either side of the EcoRI cloning site of the BAC vector pECS-BAC4. Our strategy includes the following steps: (i) generation of Southern blots with BAC clones digested with NotI and a second restriction enzyme; (ii) identification of the termini attached to the NotI/EcoRI fragment of the BAC vector via hybridization with a probe derived from sequences located between one NotI site (left or right arm) and the cloning site; (iii) ligation of the doubly digested BAC clone (NotI and the selected second restriction enzyme) with an equally doubly digested cloning plasmid vector; and (iv) confirmation of the clone as a terminus. This strategy has allowed us to begin the construction of a contig near a common bean gene that controls resistance to a group of potyviruses. PMID- 10818710 TI - Adaptation of conformation-sensitive gel electrophoresis to an ALFexpress DNA sequencer to screen BRCA1 mutations. AB - Conformation-sensitive gel electrophoresis (CSGE) has been introduced as the most reliable method for the screening of large and multi-exon genes because of its simplicity, sensitivity and specificity. Based on heteroduplex formation and with the use of mildly denaturing solvents, it allows detection of single-base mutations with accuracy. This is important in genes such as BRCA1 and BRCA2, in which alterations span the entire gene. We have adapted the CSGE assay to a fluorescent platform--a DNA sequencer one-color technology--that reduces the time involved and enhances resolving power for the complete scanning of the BRCA genes. Electrophoresis has high sensitivity and is performed in less than three hours, and the gel does not require staining with ethidium bromide. Eighteen single-base and six frameshift mutations in the BRCA1 gene were analyzed. We compared the manual and fluorescent CSGE methods, and all mutations were detected with accuracy. PMID- 10818711 TI - Cloning human natural killer cells. PMID- 10818712 TI - Generation of short-term murine natural killer cell clones to analyze Ly49 gene expression. PMID- 10818713 TI - Cloning and culturing of fetal mouse natural killer cells. PMID- 10818714 TI - Techniques for studying development of human natural killer cells and T cells. PMID- 10818715 TI - Murine natural killer cell cloning from fetal thymic organ cultures. PMID- 10818716 TI - Generation of HLA class I transfected target cell lines. PMID- 10818717 TI - Murine natural killer cells and hybrid resistance to hemopoietic cells in vivo. PMID- 10818718 TI - Hybrid resistance by mouse NK cells in vitro. PMID- 10818719 TI - Use of activated natural killer cells for tumor immunotherapy in mouse and human. PMID- 10818720 TI - Methods for in vivo analyses of natural killer (NK) cells. PMID- 10818721 TI - Assay for the transendothelial migration of human natural killer cells. PMID- 10818722 TI - Methods of analyzing chromatin changes accompanying apoptosis of target cells in killer cell assays. PMID- 10818723 TI - Methods for detecting lytic granules in natural killer cells. PMID- 10818724 TI - Fluorogenic substrates as detectors of caspase activity during natural killer cell-induced apoptosis. PMID- 10818725 TI - Techniques for studying murine natural killer cells in defense against viral infection. PMID- 10818726 TI - Assays for antibody-dependent cell-mediated cytotoxicity (ADCC) and reverse ADCC (redirected cytotoxicity) in human natural killer cells. PMID- 10818727 TI - Multiple color immunofluorescence for cytokine detection at the single cell level. PMID- 10818728 TI - Generation of antibodies to cell surface markers on mature natural killer cells. PMID- 10818729 TI - Human single-chain Fv fragments specific for natural killer cell receptors from phage display libraries. PMID- 10818730 TI - Binding of soluble KIR-Fc fusion proteins to HLA class I. PMID- 10818731 TI - Genetic analysis of a highly homologous gene family. The killer cell immunoglobulin-like receptors. PMID- 10818732 TI - Use of vaccinia virus for functional gene transfer in natural killer cells. PMID- 10818733 TI - Use of cDNA library expression cloning to identify components of heterodimeric receptor complexes. PMID- 10818734 TI - Functional analysis of natural killer cell receptors in the RNK-16 rat leukemic cell line. PMID- 10818735 TI - Use of mutant Jurkat T cell lines to define human natural killer cell surface receptor signaling pathways. PMID- 10818736 TI - Measurement of calcium mobilization responses in killer cell/target conjugates by FACS analysis. PMID- 10818737 TI - BIAcore analysis to test phosphopeptide-SH2 domain interactions. PMID- 10818738 TI - Analysis and use of mutant mice exhibiting natural killer cell defects. PMID- 10818739 TI - Isolation and analysis of natural killer cells in chickens. PMID- 10818740 TI - Analyzing nonspecific cytotoxic cells in fish. PMID- 10818741 TI - Natural killer cells and immunotoxicology. PMID- 10818742 TI - Purification of mu-calpain, m-calpain, and calpastatin from animal tissues. PMID- 10818743 TI - A simple protocol for separation and assay of mu-calpain, m-calpain, and calpastatin from small tissue samples. PMID- 10818744 TI - Purification and quantification of calcium-activated neutral proteases I and II and novel isoforms from cultured osteoblastic cells by ion-exchange fast protein liquid chromatography. PMID- 10818745 TI - Purification of calpain by affinity chromatography on reactive red-agarose or on casein-sepharose. PMID- 10818746 TI - Affinity purification of mu-calpain from erythrocytes on an immobilized peptide from the plasma membrane calcium pump. Some studies on erythrocyte mu-calpain. PMID- 10818747 TI - Expression of m-calpain in Escherichia coli. PMID- 10818748 TI - Purification and characterization of crustacean calpain-like proteinases. PMID- 10818749 TI - Drosophila calpains. Purification of a calpain-like enzyme from fruit flies, and expression in Escherichia coli. PMID- 10818750 TI - Molecular analysis of p94 and its application to diagnosis of limb girdle muscular dystrophy type 2A. PMID- 10818751 TI - Purification of recombinant calpastatin expressed in Escherichia coli. PMID- 10818752 TI - Preparation of calpastatin samples for western blotting. PMID- 10818753 TI - Isolation and characterization of calpain activator protein from bovine brain. PMID- 10818754 TI - Calpain zymography with casein or fluorescein isothiocyanate casein. PMID- 10818756 TI - Fluorescence measurements of Ca2+ binding to domain VI of calpain. PMID- 10818755 TI - Casein zymogram assessment of mu-calpain and m-calpain activity after traumatic brain injury in the rat in vivo. PMID- 10818757 TI - Kinetic analysis of human mu-calpain autolysis. PMID- 10818758 TI - A sensitive and continuous fluorometric activity assay using a natural substrate. Microtubule-associated protein 2. PMID- 10818759 TI - Measurement of calpain activity in vitro and in situ using a fluorescent compound and tau as substrates. PMID- 10818760 TI - Localization of calpain by immunofluorescence in adherent cells. PMID- 10818761 TI - A radioimmunologic technique for assessing calpain activation in cells. PMID- 10818762 TI - Calpains and myogenesis. PMID- 10818763 TI - Calpastatin (the endogenous calpain inhibitor) and membrane protein degradation in cell fusion. PMID- 10818764 TI - The role of calpain in neurofilament protein degradation associated with spinal cord injury. PMID- 10818765 TI - Calpain-mediated truncation of glutamate ionotropic receptors. Methods for studying the effects of calpain activation in brain tissue. PMID- 10818766 TI - Concurrent assessment of calpain and caspase-3 activity by means of western blots of protease-specific spectrin breakdown products. PMID- 10818767 TI - Rat renal proximal tubules, hypoxia, ionomycin, and calpain. PMID- 10818768 TI - Calpain activity in rat renal proximal tubules. An in vitro assay. PMID- 10818769 TI - Calpain activity in rat renal proximal tubules. An in situ assay. PMID- 10818770 TI - Cellular in vivo assay of calpain activity using a fluorescent substrate. Application to study of anoxic liver injury. PMID- 10818771 TI - Calpain methods in hepatic ischemia-reperfusion injury. PMID- 10818772 TI - Myocardial ischemia-reperfusion injury and proteolysis of fodrin, ankyrin, and calpastatin. PMID- 10818773 TI - Calpains in the lens and cataractogenesis. PMID- 10818774 TI - Proteolysis of cortactin by calpain in platelets and in vitro. PMID- 10818775 TI - Proteolysis of p53 protein by ubiquitous calpains. PMID- 10818776 TI - Modulation of calpain-mediated protein kinase C activation within intact cells. PMID- 10818777 TI - Strategies for regulating calpain activities in living cells. PMID- 10818778 TI - Assays of apoptosis. PMID- 10818779 TI - Tiaprofenic acid-photosensitized damage to nucleic acids: a mechanistic study using complementary in vitro approaches. AB - In order to determine whether or not tiaprofenic acid (TPA) could cause cellular DNA damage, human fibroblasts were irradiated in the presence of the drug and subsequently examined by means of the comet assay. This led to the observation that TPA actually sensitizes cellular DNA to the subsequent irradiation. When TPA was irradiated in the presence of supercoiled plasmid DNA, it produced large amounts of single-strand breaks (SSB); this is consistent with the effects observed on cellular genomic DNA by the comet assay. More importantly, low concentrations of TPA, unable to produce direct SSB, caused photo-oxidative damage to DNA as revealed by the use of excision-repair enzymes. The fact that TPA-irradiated DNA was a substrate of formamidopyrimidine glycosylase as well as endonuclease III revealed that both purine and pyrimidine bases were oxidized. This was further supported by the TPA-photosensitized oxidation of 2' deoxyguanosine which led to a product mixture characteristic of mixed type-I/II mechanisms. Thymidine was less reactive under similar conditions, but it also decomposed to give a typical type-I product pattern. Accordingly, the TPA triplet was quenched by the two nucleosides with clearly different rate constants (10(8) vs 10(7) M-1 s-1, respectively). As cellular RNA also contains oxidizable bases, it could be the target of similar processes, thus interfering with the biosynthesis of proteins by the cells. Extraction of total RNA from TPA irradiated human fibroblasts, followed by gel electrophoresis and PCR analysis, confirmed this hypothesis. Finally, photosensitization experiments with Saccharomyces cerevisiae showed that, in spite of an efficient drug-yeast interaction leading to cytotoxicity, neither intergenic recombination nor gene conversion took place. Thus, while TPA-photosensitized damage to nucleic acids can result in genotoxicity, the risk of mutagenicity does not appear to be significant. PMID- 10818780 TI - Photophysical properties of the lowest excited singlet and triplet states of thio and seleno-psoralens AB - The decay processes of the lowest excited singlet and triplet states of five heteropsoralens (HPS) were investigated by steady-state and shift-phase fluorometry and by laser-flash photolysis in different solvents. The emission spectra of HPS are detectable only in trifluoroethanol (TFE), where fluorescence lifetimes (tau F) and quantum yields (phi F) were measured. The triplet lifetimes (tau T), triplet (phi T) and singlet-oxygen production (phi delta) quantum yields were determined in benzene, ethanol and TFE by laser-flash photolysis. Semiempirical (INDO/1-CI) calculations allowed the nature of the lowest excited singlet and triplet states and transition probabilities to be obtained. Theoretical and experimental results indicate that the two lowest excited singlet states S1 and S2 of HPS are close-lying and different in nature (pi,pi* and n,pi*). The "proximity effect" between these two states controls the photophysical properties of HPS as it does for the other furocoumarins. However, HPS have a peculiar behavior with respect to the related compounds because they are fluorescent and have, in three cases, detectable intersystem crossing only in TFE. This behavior can be tentatively explained by a different energy gap and/or order between the S1 and S2 states. PMID- 10818781 TI - Systematic study of parameters influencing the action of Rose Bengal with visible light on bacterial cells: comparison between the biological effect and singlet oxygen production. AB - As part of a project to study different methods for the disinfection of effluent water, the inactivation of different microorganisms (Escherichia coli, Deinococcus radiodurans and spores of Bacillus subtilis) using a combination of a photosensitizer (Rose Bengal) with simulated sunlight and oxygen was determined under various environmental conditions (temperature, pH index). In parallel, the singlet-oxygen (1O2) production was also measured under the same conditions. Whereas the vegetative cells could be inactivated much more efficiently at increased temperature and altered index of pH, the production of 1O2 remained essentially the same under these alterations. Additionally, the relations among the sensitivities of different cell types to be killed by our photodynamic treatments (PDT) were opposite to those found after exposure to ionizing radiation. The results of photodynamic experiments do not reflect the cells' capacity to repair DNA strand breaks. Spores of B. subtilis, as a nonvegetative system, could not be inactivated by illuminations up to 100 J cm-2. Together, these findings indicate that DNA is not the primary target, the inactivation of which leads to the killing of our test organisms. Instead, the cellular envelope appears to be the component being assaulted by our PDT. PMID- 10818782 TI - Absolute rate constants for the quenching of reactive excited states by melanin and related 5,6-dihydroxyindole metabolites: implications for their antioxidant activity. AB - The triplet-excited state of benzophenone and the singlet-excited state of 2,3 diazabicyclo[2.2.2]oct-2-ene (Fluorazophore-P) have been employed as kinetic probes to obtain information on the antioxidant activity of the skin and eye pigment melanin and its biogenetic precursors 5,6-dihydroxyindole (DHI) and 5,6 dihydroxyindole-2-carboxylic acid (DHICA). The excited states were generated by the laser-flash photolysis technique and their reaction kinetics was examined by time-resolved transient absorption or fluorescence spectroscopy, respectively. The reaction between triplet benzophenone and DHI produced with unit efficiency the corresponding 6O-centered semiquinone radical, which was characterized by its characteristic transient absorption. The quenching rate constants for DHI (3.1 8.4 x 10(9) M-1 s-1) and DHICA (3.3-5.5 x 10(9) M-1 s-1) were near the diffusion controlled limit, indicating excellent antioxidant properties. Kinetic solvent effects were observed. The reactivity of synthetic melanin, assessed through the quenching rate constant of Fluorazophore-P and normalized to the number of monomer units, was more than one order of magnitude lower (2.7 x 10(8) M-1 s-1) than that of its precursors. The trend of the quenching rate constants, i.e. DHI > DHICA approximately alpha-tocopherol > melanin, along with the preferential solubility of DHICA in aqueous environments, serves to account for several experimental results from biochemical studies on the inhibition of lipid peroxidation by these natural antioxidants. PMID- 10818783 TI - Photochemical and photophysical studies of guanine derivatives: intermediates contributing to its photodestruction mechanism in aqueous solution and the participation of the electron adduct. AB - The low-intensity steady-state (254 nm), microsecond flash and nanosecond (266 nm) laser photolysis of some guanine (Gua) derivatives in aqueous solution were studied. A photodestruction yield between 10(-3) and 10(-2) at a base concentration of 75 microM was determined for 254 nm irradiation at room temperature using high-performance liquid chromatography. This yield decreases with increasing purine concentration. For a similar concentration of the purine bases (2 +/- 1) x 10(-5) M, the yield increases as follows: Gua approximately 9 ethylguanine < deoxyguanosine approximately guanosine (Guo) < guanosine 5' monophosphate. At concentrations higher than 2 x 10(-4) M the Gua derivatives' photodestruction yield seems to converge to a limiting value of the order of 10( 4). This behavior is explained in terms of self-quenching and aggregation effects which deactivate the excited states of the bases. The yields of electron photoejection have been determined in the nanosecond laser photolysis (0.083) and in the low-intensity steady-state (5.8 x 10(-3)) for Guo. Competition experiments using electron scavengers suggest that the electron adducts of the bases are one of the principal species participating in the photodestruction mechanism of these monomeric Gua. Close to 75% of the total destruction yield has contributions from initial reactions of the photojected electron at neutral pH. The quantum yield of photodestruction of Guo increases when the pH is increased as follows: 4.7 x 10( 3) (pH 1.1), 6.5 x 10(-3) (pH 2.9), 7.7 x 10(-3) (pH 7.5) and 8.1 x 10(-3) (pH 11.9). This dependence on pH and the electron scavenger experiments provide further evidence for the radical anion or its protonated form as one of the principal species involved in the photodestruction of the bases at the different pH. Under oxygen saturated conditions a 22% increase in the destruction yield is observed for Guo. However, for the dinucleotides adenylyl (3'-->5')-guanosine and thymidylyl (3'-->5')2'-deoxyguanosine, the participation of the electron is 41 and 36%, respectively, suggesting that going into a more DNA or RNA-like structure, the participation of the electron adducts species in the photodamage of DNA and RNA decreases. A mechanism of photodestruction for the Gua derivatives is proposed which takes into account these findings. PMID- 10818784 TI - Mechanism of formation of guanine as one of the major products in the 254 nm photolysis of guanine derivatives: concentration and pH effects. AB - The quantum yield of formation of guanine (Gua), one of the major products formed in the 254 nm steady-state photolysis of the following Gua derivatives (9 ethylguanine [9Et-Gua], deoxyguanosine [dGuo], guanosine [Guo], guanosine 5' monophosphate [GMP]) was determined under different conditions. The formation yield increases in the following order: 9Et-Gua < dGuo approximately Guo < GMP. Electron scavengers or triplet quenchers were incorporated into the irradiated solutions with the purpose of reacting specifically with postulated or previously identified intermediates in the photolysis of Gua derivatives. A decrease in the yield of formation of Gua is observed with increasing electron scavenger concentration or with pH. These results suggest a major contribution from Gua derivative electron adducts on the process of N(9)-R bond breakage. At an acidic pH a tautomer of the radical cation (E) of Guo is proposed as the precursor for Gua formation. The relative efficiency of the radical cation for initiating the release of free Gua depends on the pH of the solution, being less than 39% in neutral pH. Reactions from OH radicals may also result in base release as shown using N2O as additive. Finally, the formation of aggregates by the bases at concentrations used, plays an important role in the deactivation of the excited states and also in the probability of formation of the free base. PMID- 10818785 TI - Modulation of c-jun and c-fos transcription by UVB and UVA radiations in human dermal fibroblasts and KB cells. AB - We have previously demonstrated that the oxidizing component of ultraviolet-A (UVA) plays a central role in the activation of the nuclear oncogene and transcription factor, c-fos, in cultured human skin fibroblasts. We have now shown that expression of both c-jun and c-fos (AP-1) family of transcription factors is modulated by short and long wavelength solar ultraviolet (UV) radiation in human fibroblasts and human KB cells. UVA radiation activated c-jun and c-fos in both fibroblasts and KB cells, whereas ultraviolet-B (UVB) radiation activates such oncogenes only in KB cells. Moreover, decreasing the intracellular levels of reducing equivalents in human fibroblasts by glutathione (GSH) depletion lowered the UVA dose threshold for c-jun and c-fos activation several fold and greatly amplified the UVA-mediated activation of such genes. A more modest effect was observed in GSH-depleted KB cells. In both GSH-depleted fibroblasts and KB cells, UVB radiation failed to amplify c-jun and c-fos activation indicating that the oxidative component of UVB plays a minor role in the modulation of such oncogene expression. These findings clearly indicate that both c-jun and c-fos are activated by the oxidizing component of UVA radiation in human fibroblasts and KB cells, while UVB-mediated modulation seems to be restricted to human epithelial cells and does not involve oxidizing intermediates. PMID- 10818786 TI - Continuing damage to rat retinal DNA during darkness following light exposure. AB - The damaging effects of visible light on the mammalian retina can be detected as functional, morphological or biochemical changes in the photoreceptor cells. Although previous studies have implicated short-lived reactive oxygen species in these processes, the termination of light exposure does not prevent continuing damage. To investigate the degenerative processes persisting during darkness following light treatment, rats were exposed to 24 h of intense visible light and the accumulation of DNA damage to restriction fragments containing opsin, insulin 1 or interleukin-6 genes was measured as single-strand breaks (ssb) on alkaline agarose gels. With longer dark treatments all three DNA fragments showed increasing DNA damage. Treatment of rats with the synthetic antioxidant dimethylthiourea prior to light exposure reduced the initial development of alkali-sensitive strand breaks and allowed significant repair of all three DNA fragments. The time course of double-strand DNA breaks was also examined in specific genes and repetitive DNA. Nucleosomal DNA laddering was evident immediately following the 24 h light treatment and increased during the subsequent dark period. The increase in the intensity of the DNA ladder pattern suggests a continuation of enzymatically mediated apoptotic processes triggered during light exposure. The protective effects of antioxidant suggests that the light-induced DNA degradative process includes both early oxidative reactions and enzymatic processes that continue after cessation of light exposure. PMID- 10818787 TI - The effects of epimerization at the 3(1)-position of bacteriochlorophylls c on their aggregation in chlorosomes of green sulfur bacteria. Control of the ratio of 3(1) epimers by light intensity. AB - R- and S-epimerization at the 3(1) position of bacteriochlorophyll (BChl) c and the formation of rod-like aggregates in chlorosomes of green sulfur bacteria were markedly affected in Chlorobium (Cb.) tepidum and Cb. limicola by cultivation under various light intensities (photon fluence rate). The stronger the light, the higher the ratio of the S-epimer to the R-epimer for each homolog of BChl c in the bacteria. S[P,E] BChl cF and S[I,E] BChl cF were found to be the major S epimers in Cb. tepidum and Cb. limicola, respectively. R[P,E] BChl cF decreased markedly compared to R[E,E] BChl cF in Cb. tepidum, whereas no observable change in the ratio of R[P,E]/R[E,E] was detected for Cb. limicola. With increase in light intensity the Qy absorption maximum of the bacteria shifted to shorter wavelengths. In vitro spectroscopic studies of the aggregates showed a marked difference in the formation of aggregates from R- and S-epimers of BChl c; the S epimers formed aggregates much more slowly than did the R-epimers. These results suggest that the ratio of the epimers of BChl c might significantly affect the aggregation of BChl in the chlorosome. We propose different roles for the R- and S-epimers in chlorosomes of Cb. limicola and Cb. tepidum. PMID- 10818788 TI - Assignment of the lowest Qy-state and spectral dynamics of the CP29 chlorophyll a/b antenna complex of green plants: a hole-burning study. AB - Low-temperature absorption, fluorescence and persistent non-photochemical hole burned spectra are reported for the CP29 chlorophyll (Chl) a/b antenna complex of photosystem II of green plants. The absorption-origin band of the lowest Qy-state lies at 678.2 nm and carries a width of approximately 130 cm-1 that is dominated by inhomogeneous broadening at low temperatures. Its absorption intensity is equivalent to that of one of the six Chl a molecules of CP29. The absence of a significant satellite hole structure produced by hole burning, within the absorption band of the lowest state, indicates that the associated Chl a molecule is weakly coupled to the other Chl and, therefore, that the lowest-energy state is highly localized on a single Chl a molecule. The electron-phonon coupling of the 678.2 nm state is weak with a Huang-Rhys factor S of 0.5 and a peak phonon frequency (omega m) of approximately 20 cm-1. These values give a Stokes shift (2S omega m) in good agreement with the measured positions of the absorption band at 678.2 nm and a fluorescence-origin band at 679.1 nm. Zero-phonon holes associated with the lowest state have a width of approximately 0.05 cm-1 at 4.2 K, corresponding to a total effective dephasing time of approximately 400 ps. The temperature dependence of the zero-phonon holewidth indicates that this time constant is dominated at temperatures below 8 K by pure dephasing/spectral diffusion due to coupling of the optical transition to the glass-like two-level systems of the protein. Zero-phonon hole-widths obtained for the Chl b bands at 638.5 and 650.0 nm, at 4.2 K, lead to lower limits of 900 +/- 150 fs and 4.2 +/- 0.3 ps, respectively, for the Chl b-->Chl a energy-transfer times. Downward energy transfer from the Chl a state(s) at 665.0 nm occurs in 5.3 +/- 0.6 ps at 4.2 K. PMID- 10818789 TI - Heterodimeric versus homodimeric structure of the primary electron donor in Rhodobacter sphaeroides reaction centers genetically modified at position M202. AB - Using light-induced Fourier-transform infrared (FTIR) difference spectroscopy of the photo-oxidation of the primary donor (P) in chromatophores from Rhodobacter sphaeroides, we examined a series of site-directed mutants with His M202 changed to Gly, Ser, Cys, Asn or Glu in order to assess the ability of these side chains to ligate the Mg atom of one of the two bacteriochlorophylls (BChl) constituting P. In the P+QA-/PQA FTIR difference spectra of the mutants HG(M202), HS(M202), HC(M202) and HN(M202), the presence of a specific electronic transition at approximately 2650-2750 cm-1 as well as of associated vibrational (phase-phonon) bands at approximately 1560, 1480 and 1290 cm-1 demonstrate that these mutants contain a BChl/BChl homodimer like that in native reaction centers with the charge on P+ shared between the two coupled BChl. In contrast, the absence of all of these bands in HE(M202) shows that this mutant contains a BChl/bacteriopheophytin heterodimer with the charge localized on the single BChl, as previously determined for the mutant HL(M202). Furthermore, the spectra of the heterodimers HE(M202) and HL(M202) are very similar in the 4000-1200 cm-1 IR range. Perturbations of the 10a-ester and 9-keto carbonyl modes for both the P and P+ states are observed in the homodimer mutants reflecting slight variations in the conformation and/or in position of P. These perturbations are likely to be due to a repositioning of the dimer in the new protein cavity generated by the mutation. PMID- 10818790 TI - Resonance Raman and surface-enhanced resonance Raman spectra of LH2 antenna complex from Rhodobacter sphaeroides and Ectothiorhodospira sp. excited in the Qx and Qy transitions. AB - Well-resolved vibrational spectra of LH2 complex isolated from two photosynthetic bacteria, Rhodobacter sphaeroides and Ectothiorhodospira sp., were obtained using surface-enhanced resonance Raman scattering (SERRS) exciting into the Qx and the Qy transitions of bacteriochlorophyll a. High-quality SERRS spectra in the Qy region were accessible because the strong fluorescence background was quenched near the roughened Ag surface. A comparison of the spectra obtained with 590 nm and 752 nm excitation in the mid- and low-frequency regions revealed spectral differences between the two LH2 complexes as well as between the LH2 complexes and isolated bacteriochlorophyll a. Because peripheral modes of pigments contribute mainly to the low-frequency spectral region, frequencies and intensities of many vibrational bands in this region are affected by interactions with the protein. The results demonstrate that the microenvironment surrounding the pigments within the two LH2 complexes is somewhat different, despite the fact that the complexes exhibit similar electronic absorption spectra. These differences are most probably due to specific pigment-pigment and pigment-protein interactions within the LH2 complexes, and the approach might be useful for addressing subtle static and dynamic structural variances between pigment-protein complexes from different sources or in complexes altered chemically or genetically. PMID- 10818791 TI - Structural transformation among the aggregate forms of bacteriochlorophyll c as determined by electronic-absorption and NMR spectroscopies: dependence on the stereoisomeric configuration and on the bulkiness of the 8-C side chain. AB - Transformation among the aggregate forms of bacteriochlorophyll (BChl) c characterized by the wavelength of the Qy absorption, i.e. the dimer (B675), B705, B720 and B745, was traced by electronic-absorption spectroscopy for each of the isomers including R[E,E], R[P,E], R[I,E], S[P,E] and S[I,E] suspended in the mixtures of methylene chloride and n-hexane. A combination of NMR spectroscopy determining the structural motifs and calculation of the shift of the Qy absorption reflecting the long-range transition dipole-transition dipole interactions among the macrocycles in the entire aggregate structures proposed the following models: B705d (B705d'), a linear array of straight (inclined) columns consisting of a pair of the piggyback dimers; B720d and B745d, an assembly of two and five shifted-inclined columns consisting of more than six piggyback dimers; and B720m and B745m, an assembly of one and two parallel stepwise stacking of approximately 30 monomers. Calculations of the steric energies rationalized two different pathways of transformations: the dimer- >B705d (B705d')-->B720d-->B745d for the R isomers; and the monomer-->(B720m)- >B745m for the S isomers. Addition of S[I,E] seems to trigger the B745d-->B745m transformation of the R isomers. PMID- 10818792 TI - Light scattering and absorption model for the quantitative interpretation of human blood platelet spectral data. AB - Multiwavelength ultraviolet-visible (UV-Vis) transmission spectroscopy is a relatively simple technique that can provide considerable quantitative information on the properties of micron and submicron particle suspensions. Two important particle properties are particle size distribution (PSD) and chemical composition. These properties provide characteristics for the identification and classification of biological systems ranging in size and composition from proteins and nucleic acids to cells. By measuring the complete UV-Vis spectrum, the combined scattering and absorption properties are obtained as a function of wavelength. The quantitative evaluation of the size distribution and chemical composition is accomplished through the application of light-scattering theory. This paper reports on the estimation of the optical properties of human blood platelets and their use in the interpretation of platelet UV-Vis spectra within the context of Mie theory. The model developed herein provides reliable and accurate estimates for the PSD and particle number of platelet suspensions. One potential application of this characterization method is in the analysis of platelet activation by thrombin. Quantification of spectral data with respect to average particle size and particle number provides a real-time description of the dramatic changes that accompany the platelet activation process. PMID- 10818793 TI - Induction of multinucleated cells caused by UVA exposure in different stages of the cell cycle. AB - Fibroblasts of the line 3T3 from swiss albino mice were exposed to ultraviolet A (UVA) irradiation. The cells were synchronized by treatment with nocodazole and mitotic shake-off, and then exposed to UVA irradiation in different stages of the cell cycle. Their photosensitivity varied through the cell cycle, being greatest in the G2 phase. UVA irradiation was found to induce the formation of multinucleated cells. Cells in the G1 phase were found to be most prone to multinucleation 15 min after UVA irradiation, while cells exposed to UVA irradiation in S and G2 phases contained the largest fractions of multinucleated cells 24 h after treatment. The present results indicate that multinucleated cells are formed by fusion of two or more cells shortly after UVA irradiation of early G1 cells, while impairment of cytokinesis is a possible explanation for the delayed formation of multinucleated cells after irradiation in S and G2. PMID- 10818794 TI - Effects of photoinduced membrane rigidification on the lysosomal permeability to potassium ions. AB - Mechanism for the photoinduced increase in the lysosomal K+ permeability is still unknown. In this study, we investigated the effect of photodamage-induced membrane rigidification on the lysosomal K+ permeability by measuring the membrane potential with bis(3-propyl-5-oxoisoxazol-4-yl)pentamethine oxonol and by monitoring proton leakage with p-nitrophenol. Membrane fluidity was measured by the steady-state fluorescence anisotropy of 1,6-diphenyl-1,3,5-hexatriene. Methylene blue-mediated photodamage to lysosomes decreased their membrane fluidity and increased their K+ permeability. The photoinduced increase in the K+ permeability can be reversed by fluidizing the rigidified lysosomal membranes with benzyl alcohol. The results suggest that the membrane rigidification induced by photodamage may increase lysosomal K+ permeability. This conclusion is supported by the observation that rigidifying lysosomal membranes by the treatment with membrane rigidifier cholesteryl hemisuccinate also enhanced the lysosomal K+ permeability. PMID- 10818795 TI - Quantitative analysis of Pc 4 localization in mouse lymphoma (LY-R) cells via double-label confocal fluorescence microscopy. AB - Photodynamic therapy (PDT) is a novel cancer therapy that uses light-activated drugs (photosensitizers) to destroy tumor tissue. Reactive oxygen species produced during PDT are thought to cause the destruction of tumor tissue. However, the precise mechanism of PDT is not completely understood. To provide insight into the in vitro mechanisms of PDT, we studied the subcellular localization of the photosensitizer HOSiPcOSi(CH3)2-(CH2)3N(CH3)2 (Pc 4) in mouse lymphoma (LY-R) cells using double-label confocal fluorescence microscopy. This technique allowed us to observe the relative distributions of Pc 4 and an organelle-specific dye within the same cell via two, spectrally distinct, fluorescence images. To quantify the localization of Pc 4 within different organelles, linear correlation coefficients from the fluorescence data of Pc 4 and the organelle-specific dyes were calculated. Using this measurement, the subcellular spatial distributions of Pc 4 could be successfully monitored over an 18 h period. At early times (0-1 h) after introduction of Pc 4 to LY-R cells, the dye was found in the mitochondria, lysosomes and Golgi apparatus, as well as other cytoplasmic membranes, but not in the plasma membrane or the nucleus. Over the next 2 h, there was some loss of Pc 4 from the lysosomes as shown by the correlation coefficients. After an additional incubation period of 2 h Pc 4 slowly increased its accumulation in the lysosomes. The highest correlation coefficient (0.65) was for Pc 4 and BODIPY-FL C5 ceramide, which targets the Golgi apparatus, and also binds to other cytoplasmic membranes. The correlation coefficient was also high (0.60) for Pc 4 and a mitochondria-targeting dye (Mitotracker Green FM). Both of these correlation coefficients were higher than that for Pc 4 with the lysosome-targeting dye (Lysotracker Green DND-26). The results suggest that Pc 4 binds preferentially and strongly to mitochondria and Golgi complexes. PMID- 10818796 TI - 5-aminolevulinic acid, but not 5-aminolevulinic acid esters, is transported into adenocarcinoma cells by system BETA transporters. AB - 5-aminolevulinic acid (5-ALA) and its ester derivatives are used in photodynamic therapy as precursors for the formation of photosensitizers. This study relates to the mechanisms by which 5-ALA is transported into cells. The transport of 5 ALA has been studied in a human adenocarcinoma cell line (WiDr) by means of [14C] labeled 5-ALA. The rate of uptake was saturable following Michaelis-Menten kinetics (K(m) = 8-10 mM and Vmax = 18-20 nmol.(mg protein x h)-1), and Arrhenius plot of the temperature-dependent uptake of 5-ALA was characterized by a single discontinuity at 32 degrees C. The activation energy was 112 kJ.mol-1 in the temperature range 15 degrees-32 degrees C and 26 kJ.mol-1 above 32 degrees C. Transport of 5-ALA was Na+ and partly Cl(-)-dependent. Stoichiometric analysis revealed a Na+:5-ALA coupling ratio of 3:1. With the exception of valine, methionine and threonine, zwitterionic and basic amino acids inhibited the transport of 5-ALA. 5-ALA methyl ester was not an inhibitor of 5-ALA uptake. The transport was most efficiently inhibited, i.e. by 65-75%, by the beta-amino acids, beta-alanine and taurine and by gamma-aminobutyric acid (GABA). Accordingly, 5-ALA, but not 5-ALA methyl ester, was found to inhibit cellular uptake of [3H]-GABA and [14C]-beta-alanine. Protoporphyrin IX (PpIX) accumulation in the presence of 5-ALA (0.3 mM) was attenuated 85% in the presence of 10 mM beta-alanine, while PpIX formation in cells treated with 5-ALA methyl ester (0.3 mM) or 5-ALA hexyl ester (4 microM) was not significantly influenced by beta alanine. Thus, 5-ALA, but not 5-ALA esters, is transported by beta-amino acid and GABA carriers in this cell line. PMID- 10818797 TI - Luciferase and urate may act as antioxidant defenses in larval Pyrearinus termitilluminans (Elateridae: Coleoptera) during natural development and upon 20 hydroxyecdysone treatment. AB - Insects show unique adaptations to cope with oxidative challenges during larval development, metamorphosis and adulthood. Our previous findings suggested that bioluminescence may act as an auxiliary oxygen-detoxifying mechanism in larvae of Pyrearinus termitilluminans (Elateridae: Coleoptera). We now study the antioxidant status in larval P. termitilluminans, evaluated in terms of levels of chemical and enzymatic antioxidant defenses, as compared to luciferase activity in the prothorax (intensely bright) and abdomen (dim) of the larvae, during natural- and 20-hydroxyecdysone (20-HE)-induced development. In the prothorax, relative total SOD activities in small (< 1 cm), medium (1-2 cm) and large (> 2 cm) larvae were 1.00:0.53:0.32. Catalase activity also decreased with development (1.00:0.69:0.55). In contrast, prothorax luciferase activities and urate content increased with ratios of 1.0:2.2:2.5 and 1:15:97, respectively. No increases were found in the level of prothorax lipid and protein oxidation. In the abdomen, luciferase activity decreased markedly with development (1.00:0.33:0.17), as did other antioxidant enzymes, including dehydroascorbate reductase (1.00:0.59:0.17) and levels of lipid peroxidation products and protein carbonyls. Similar variations were observed in antioxidant enzyme activities when the larvae were treated with 20-HE, except for prothorax catalase. As observed in natural larval growth, luciferase activity was augmented (two-fold in prothorax) upon steroid treatment, and the levels of thiobarbituric acid-reactive substances were magnified in both segments. The increase of luciferase activity and a higher urate content in the prothorax during larval development may reflect metabolic adaptations to keep levels of oxyradicals low in order to compensate for decreased antioxidant enzyme activities. PMID- 10818799 TI - Characterization of blue-light and developmental regulation of the photolyase gene phr1 in Trichoderma harzianum. AB - Blue light and development regulate the expression of the phr1 gene of the filamentous fungus Trichoderma harzianum. The predicted product of phr1, the DNA repair enzyme photolyase, is likely to help protect Trichoderma, which grows in the soil as a mycoparasite or saprophyte, from damage upon emergence and exposure to ultraviolet-c. phr1 is transiently expressed in mycelium and conidiophores after illumination. phr1 mRNA also accumulates in conidiophores during development and spore maturation. As no other genes displaying rapid, direct light regulation have been described previously in this organism, we have characterized the fluence and time dependence of phr1 induction, and its relation to sporulation and photoreactivation. Induction is transient following a pulse, and, with slower decay, in continuous light. This implies that the photoreceptor, transducers or response are capable of adaptation. About two-fold more light is required to induce phr1 than conidiation, but this difference is modest, so both responses could use the same or similar chromophore. Adenosine 3':5'-cyclic monophosphate bypasses the requirement for light for sporulation, while atropine prevents sporulation even after photoinduction. Light regulation of phr1, however, is indifferent to both these effectors. Induction of photolyase expression behaves as a direct, rapid response to light, independent of the induction of sporulation. Indeed, illumination of mature spores increases their capacity for photoreactivation. Blue light seems to warn the organism against the harmful effects of short wave-lengths, inducing phr1 expression and sporulation by pathways that are, at least in part, distinct. PMID- 10818798 TI - The photoreactions of recombinant phytochrome from the cyanobacterium Synechocystis: a low-temperature UV-Vis and FT-IR spectroscopic study. AB - The interconvertible photoreactions of recombinant phytochrome from Synechocystis reconstituted with phycocyanobilin were investigated by light-induced optical and Fourier-transform infrared (FT-IR) difference spectroscopy at low temperatures for the first time. The photochemistry was found to be deferred below -100 degrees C for the transformation of red-absorbing form of phytochrome (Pr)-->far red-absorbing form of phytochrome (Pfr), and no formation of an intermediate similar to the photoproduct of phytochrome A obtained at -140 degrees C (lumi-R) was observed. Two intermediates could be stabilized below -40 degrees C and between -40 and -20 degrees C, and were denoted as meta-Ra and meta-Rc, respectively. Above -20 degrees C Pfr was obtained. In the reverse reaction two intermediates could be stabilized below -60 degrees C (lumi-F) and between -60 and -40 degrees C (meta-F). The FT-IR difference spectra of the late Pr-->Pfr photoreaction show great similarities to the spectra obtained from oat phytochrome A suggesting similar conformation of the chromophore and interactions with its protein environment, whereas deviations in the spectra of meta-Ra were observed. A large band around 1700 cm-1 in the difference spectra between the intermediates and Pr which is tentatively assigned to the C19=O group of the prosthetic group indicates the Z,E isomerization around the C15=C16-methine bridge of the chromophore during the formation of meta-Ra. In the difference spectra of the parent states only small differences are observed in this region suggesting that the frequency of the carbonyl group is similar in Pr and Pfr. Since the FT-IR difference spectra between lumi-F and Pfr show great similarities to the spectra of the parent states, it is assumed that during the formation of lumi-F the chromophore largely returns into the primary Pr conformation. The FT IR spectra recorded in a medium of 2H2O generally show a downshift of the significant bands due to the isotope effect. The appearance of a characteristic band around 935 cm-1 in all 2H2O spectra suggests an assignment to an N-2H bending vibration of the chromophore. PMID- 10818801 TI - ["La Revista de Investigacion Clinica" facing new challenges and old conflicts]. PMID- 10818800 TI - Immunological and biochemical evidence that blepharismin is not a prosthetic group. AB - A polyclonal, multispecific antiserum was raised against a whole 3[(3 cholamidopropyl)-dimethylammonio]-1-propanesulfonate-extract of nonirradiated Blepharisma japonicum cells. It was used to reveal the composition of solutions that were hypothesized to contain the photoreceptor of the ciliate. A Bio-Gel A 1.5 m fine column chromatography of the extract allowed recovery of a single elution peak isolated by recording the 580 nm light absorbance. Fused-rocket immunoelectrophoresis of this material revealed a large number of > 300 kDa coeluted proteins. Blepharismin-rich material with a molecular mass of approximately 50 kDa, consisting of at least nine proteins was obtained when the same extract underwent preparative isoelectric focusing before column chromatography separation. Purification of the pigment obtained from light exposed cells gave blepharismin-rich material with a molecular weight of approximately 200 kDa. Comparison of the materials obtained under the same conditions, either from the dark-kept or light-irradiated cells, by means of pore gradient electrophoresis confirmed that proteins present in the two preparations were different. It revealed only a very small amount, if any, of proteins in the chromatography fractions with the highest absorbance at 600 nm. Results are discussed on the basis of the hypothesis that a specific blepharismin-binding protein does not exist in the protozoan. PMID- 10818802 TI - [Primary gastric lymphoma in Mexico: annual cure of about 200 patients with antibiotic treatment?]. PMID- 10818803 TI - [Are diagnoses done with fine needle aspiration biopsy reliable for breast nodules?]. PMID- 10818804 TI - [New evidence for the usefulness of an old drug in the treatment of heart failure]. PMID- 10818805 TI - [Tolerance induction in transplant recipients: are we getting closer to our goal?]. PMID- 10818806 TI - Increasing frequency of gastric lymphoma in two national institutes of health in Mexico. AB - BACKGROUND: Some studies performed in developed countries with caucasian population have suggested an increase in the frequency of primary gastric lymphoma (PGL) in the past two decades. OBJECTIVE: To establish the frequency of PGL in two National Institutes of Health in Mexico in an interval of 18 years. METHODS: We reviewed 1,854 gastric malignant neoplasms diagnosed between 1979 1996 in mestizo patients seen at two different institutions. One of them (INC) is an oncological hospital, whereas the other one (INN) is a general hospital which treats patients with benign as well as malignant gastrointestinal diseases. RESULTS: A hundred and sixty three (8.8%) gastric lymphomas were identified from 1,854 gastric malignant tumors. The incidence of PGL showed a two to three-fold increase in the last years of the study in both institutions. The increased frequency was highly significant when compared to the initial rate. This increase does not appear to be due to patient selection criteria nor to the number of malignant gastric tumors in our institutions. CONCLUSIONS: Our results indicate an increased frequency of PGL the patient population of two National Institutes of Health in Mexico during the last years. Additional studies are necessary in order to define in the increase in PGL is limited to certain geographical areas or ethnic groups. PMID- 10818807 TI - [Scintigraphy with erythrocytes labelled in vitro with 99mTc compared with 111In octreotide in the detection of carotid glomus]. AB - OBJECTIVE: Demonstrate the utility of centelleography using 111In-octreotide in carotid glomus simple o multiple in comparison to erythrocytes with 99mTc calculate the sensitivity, specificity and accuracy of both test. DESIGN: Comparative survey (Diagnostic test) Place: Hospital Oncologico de Concentracion del Distrito Federal (Mexico). SUBJECTS: Twenty patients sent to nuclear medicine service for carotid glomus diagnosis. MAIN MEASURES: All of the patients went under a centelleography with 111In-octreotide labeled erythrocytes. Diagnostic was confirmed with US and CAT, taking as the gold standard the selective angiography and/or histopathology. RESULTS: Using 99mTc-labeled erythrocytes resulted in; True positives 13, true negatives 3, false positive 3, false negative 1. Sensitivity 92%, specificity 50%, accuracy 80% and Using 111In octreotide: True positive 14, true negative 6, and 100% sensitivity, specificity was obtained. CONCLUSIONS: We consider that Centelleography with 111In-octreotide must be the selected image methodology to accurately diagnose carotid glomus, previous to surgery, or in case of patients prior to radiotherapy that because of age, damage size or location can not be candidates for surgery. PMID- 10818809 TI - Parenteral infusions as culture media from a viewpoint of nosocomial bacteremia. AB - OBJECTIVE: To assess the growth patterns of selected organisms in common parenteral solutions, in order to ascertain implications for nosocomial bacteremia. DESIGN: A microbial suspension of approximately 300 CFU/mL was sequentially inoculated into common parenteral infusions from three different manufacturers and incubated at room temperature. Initially, 11 bacterial isolates and one Candida species from clinical specimens were studied. Eight gram-negative rods (GNR) were tested at varying pH's. Species variability was examined by testing an additional 39 isolates. RESULTS: The eight GNR grew in Ringer's lactate (RL) from two manufacturers and only two grew in dextrose 5% in water (D5/W) (Klebsiella pneumoniae and Serratia marcescens). No organism grew in saline or dextrose 5% in saline. The gram-positive cocci and Candida did not grow in any solution. No significant changes in growth were found after modifying the pH of solutions. Significant inter- and intra-species growth variability was noted. CONCLUSIONS: RL is a good culture media for GNR and D5/W is a poor culture media with the exception of some bacteria of the Tribe Klebsielleae. We recommend to follow high standards of nursing practice for administering intravenous infusions and to avoid nutrient-containing solutions for prolonged parenteral use, when possible. PMID- 10818808 TI - [Continuous hemodialysis in the treatment of acute kidney failure]. AB - INTRODUCTION: Continuous replacement therapy of renal function has gained acceptance over the last decade for the treatment of acute renal failure. In the present study we present our experience using continuous hemodialysis (CHD) in our institution. PATIENTS AND METHODS: This is a prospective analysis of the CHD treated patients in the intensive care unit (ICU) of our institution over an 24 month period. CHD was performed through a double-lumen catheter such as Mahurkar. We have performed 28 CHD procedures in 28 patients, from which four were excluded from the analysis. Three patients were excluded as CHD lasted less than 12 hours and one patient because he had chronic renal failure. The studied variables were: heart and respiratory rate, mean arterial pressure, body temperature, APACHE II classification status, arterial gasometry, cell blood count, BUN, creatinine, serum electrolytes, and hepatic enzymes. We also registered urine output, diuretic use, and the mean dose of inotropic drugs employed per day. These variables were obtained at the admittance to the ICU, before the initiation of CHD and after 24 and 48 hours. We also registered age, gender, and final evolution. RESULTS: We evaluated 24 patients with mean age of 58.1 +/- 17.5 years in which CHD was use for a mean time of 4.6 +/- 2.8 days. Total ultrafiltrate was 19.5 +/- 8.4 liters, for a mean of 4.2 liters per day. CHD resulted in improvement of heart and respiratory rate, mean arterial pressure and laboratory variables such as arterial pH, bicarbonate concentration, BUN and potassium. It also decreased significant by the use of inotropic drugs. Five out of twenty-four patients survived (20.8%). The survived patients had significant lower age than the died patients (39.2 +/- 20 years vs. 63 +/- 13.3; p < 0.001), lower time between the admittance to ICU and the beginning of CHD (1.4 +/- 0.5 days vs. 3.5 +/- 2.6; p < 0.01) and lower APACHE II classification at admittance to ICU (7.4 +/- 1.6 vs. 19.0 +/- 2.7; p < 0.001) and at the start of CHD (13.6 +/- 3.2 vs. 24.7 +/- 3.7; p < 0.001). However, multivariate analysis revealed that the only variable associated with a better survival was a lower time between the admittance to intensive care and the beginning of CHD. DISCUSSION: CHD is a safe technique that can be used for acute renal failure patients who have contraindications for intermittent HD. This technique can be used in hospitals offering intermittent hemodialysis and intensive care. CHD use is associated with improvement of hemodynamic and metabolic alterations in patients with shock. Our data support the concept that the earlier the initiation of CHD the better the prognosis. PMID- 10818810 TI - [Validation and reliability of the 1,000 meter aerobic test]. AB - BACKGROUND: In humans, measurement of maximum oxygen uptake is important because it is related to cardiovascular health. In Mexico there is scanty information in nationals and unfortunately such a data has been gathered from laboratory and field indirect methods ignoring reliability, validation and cross validation. PURPOSE: This study was designed to determine the validity and reliability of the 1,000 meters walk-run test in adults. METHODS: Fifty-one healthy subjects (31 men and 20 women) participated in four maximal tests; two in a laboratory on a treadmill for direct measurement of VO2max (L1 and L2) and twice in a track for distance walk-run time in 1,000 meters (C1 and C2). Values were analyzed by interclass correlation, Pearson product moment and "t" test. RESULTS: The VO2max in L1 y L2 were 2.67 and 2.70 L.min-1, (R = 0.97) and 44.2 and 46.2 mL.kg-1.min 1, meanwhile VO2max estimated from field were 44.2 and 46.2 mL.kg-1.min-1, (R = 0.98). Pearson correlation obtained from all subjects between laboratory and field test was of r = 0.88, without significant difference (P > 0.05). From those data we development the following equation: VO2max = 71.66-5.85 (t), where 71.66 and 5.85 are constants and t is the time employ in performing the test, R = 0.86, (p < 0.05). CONCLUSION: The 1,000 meters distance walk-run for time is highly reliable and valid test for estimation of VO2max in Mexican people. The test is safe, easy and short and can be administrated with a minimal amount of equipment. We propose its utilization in other population in order to perform cross validation. PMID- 10818811 TI - [Our editing work and analysis of peer review in 1994-98]. PMID- 10818812 TI - [Physiopathology and treatment of acquired immunodeficiency syndrome]. AB - The last pandemia of the century, that of the Acquired Immunodeficiency Syndrome (AIDS) caused by the Human Immunodeficiency Virus (HIV), continues to advance with gigantic steps at about 18,000 new infections per day worldwide. In the last four years, breakthroughs have been achieved and new medications have been introduced, which have impacted the progression of HIV and its disease, decreasing associated morbidity and mortality. During the course of infection, HIV replicates actively producing as much as 10(10) genetically different virions (quasi-species), which relates to immune escape, higher pathogenicity and drug resistance. Persistent viral replication causes T CD4+ cell destruction and immunodeficiency through several mechanisms. Currently, there are 14 approved anti-retrovirals that when used as triple regimens have been able to decrease opportunistic infections, hospitalization and mortality rates. Unfortunately, these regimens still have many limitations, do not cure and can only suppress the virus effectively in 50% of the treated patients. Besides, when they are inadequately used there is associated resistance development. On the other hand, indications for treatment initiation are changing continuously and heading towards a conservative approach. In the case of salvage regimens, there are only general guidelines that have not been evaluated in clinical studies. Resistance assays have great limitations and their use is very specific. All these factors have made antiretroviral treatment a very complicated issue that should be prescribed and followed by an expert. PMID- 10818813 TI - [Antimicrobial resistance in tuberculosis: mechanisms and therapeutic options]. PMID- 10818814 TI - [Pathogenesis of myasthenia gravis]. AB - Myasthenia gravis is a neuromuscular, autoimmune, and acquired disturbance characterized by weakness and fatigue of skeletal muscles. During the past two decades, remarkable progress has been made in the understanding of myasthenia gravis, and the new knowledge has been applied directly to the clinical diagnosis and treatment of this formerly severe disease. Myasthenia gravis is undoubtedly the most thoroughly understood of all human autoimmune diseases and has served as a model for the elucidation of mechanisms underlying other autoimmune disorders. In this review we mention the most important physiopathological aspects and its application in the clinic practice. PMID- 10818815 TI - [Thrombosis, antibodies, and systemic lupus erythematosus]. PMID- 10818816 TI - [From temples and sanctuaries to hospitals; 6,000 years of history]. AB - OBJECTIVE: To find out when and where the hospitals began, its history and evolution. MATERIAL: Review of books and Journals of personal and Instituto Nacional de la Nutricion Salvador Zubiran, libraries about history of medicine and surgery. RESULTS: During the last 5000 years the history of man has been full of events, the beginning and development of hospitals has been one of them. Everything started in Sumeria in 3500 bC, some changes were developed in Egypt, Greece and Rome. In the 4th century aD the first true hospitals are opened. Since then the hospitals had improved until now. CONCLUSIONS: The sick man has been looking for the healing of his ailments, and few places made him feel protected as the hospital does. The birth, growing and evolution of such institutions had been slow and steady until the end of the second millennium aD. PMID- 10818817 TI - [Diffuse pulmonary calcinosis in a case of primary hyperparathyroidism]. PMID- 10818818 TI - Proceedings of the workshop on possible health implications of the dioxin crisis. PMID- 10818819 TI - [Animal experiments: legal, scientific and ethical aspects]. AB - Among the legal aspects the following topics are treated: the definitions of an experimental animal, an animal experiment and alternative methods with special reference to the 3 R's (replacement, reduction and refinement of animal experiments); the qualifications, education and training of researchers and animal technicians; the licence for animal experimentation; the control on animal welfare; the origin and identification of experimental animals; statistical data on the number of experimental animals; ethics committees and their structure and functions in The Netherlands and Flanders. Extrapolation, species specificity and variability are the most important scientific limitations of animal experimentation. After a short historical survey on the man-animal relation, the following ethical aspects are discussed: the instrumental versus intrinsic value of an experimental animal; the hybrid status of the animal; the objectives of animal rights movements; the balance between the human benefit of an animal experiment and the discomfort for the animal; the problem of animal rights and animal suffering and pain. PMID- 10818820 TI - [The use of laboratory animals or alternatives in fundamental research]. AB - The use of animals in fundamental and applied research has become a point of controversy. Animals were first used for food or as pets and a historical overview is given. Research was applied or empirical. Fundamental research only developed in the 18th century, without much consideration for animal welfare. It is concluded that even now the use of animals for research remains necessary. However, one should also use alternative methods as often as possible, taking into consideration the three R's: reduction, refinement and replacement. We would like to add the use of comparative research in order to look for animal models in lower vertebrates. PMID- 10818821 TI - Induction of xenotransplant tolerance. AB - Clinical xenotransplantation will depend on the induction of xeno-tolerance. Conversely, xenotransplantation may offer opportunities to induce transplantation tolerance. We have previously shown that, xenotransplant tolerance for vascularized hamster organs could be achieved in athymic rats as far as the T cell independent xeno-reactivity is concerned. This tolerance was shown to be based on specific T-independent B lymphocyte and NK cell unresponsiveness. In the present study we have shown that this T-independent xeno-tolerance can be achieved also in a semi-discordant situation using rat recipients with high titers of pre-existing anti-hamster IgM xenoantibodies. In addition, we showed that T-independent xeno-tolerance can also be induced together with T-dependent xeno-tolerance using xeno-thymus transplantation. These experiments may be of relevance for clinical xenotransplantation. The major next question to be addressed is to see how self tolerance in xeno-thymus grafted recipients can be improved as until now the latter recipients usually develop a multi-organ autoimmune syndrome, several weeks after transplanting a xeno-thymus. PMID- 10818822 TI - [Latex allergy and cross-reactions: a new threat?]. AB - Allergy to latex proteins has become an important problem, especially in the medical profession, but also in people who have regularly contact with latex products. The responsible antigens are proteins, which are present in the natural rubber latex (NRL), extracted from Hevea brasiliensis. These proteins are important in the synthesis of rubber and in the defence of the plant against external noxes (pathogenesis related proteins). Life threathening reactions occur more frequently in the operating theatre. Sensitisation can occur via different routes (aerogen, skin, parenteral, ...) but aerogenic sensitisation seems to be very important. Starch particles are important vectors of the antigens. Diagnosis is made via history, confirmed by demonstration of specific IgE via skin test (in vivo) or via an in vitro method (e.g. CAP, ELISA). Immunological cross reactions are very frequent with other plant or fruit allergens but cross allergy is less frequent (sensitivity and specificity of the in vitro test varies considerably depending on the nature of the fruit or vegetable investigated). There is also a cross reactivity with Ficus. Prevention with latex allergens becomes primordial in hospital settings. The industry is aware of the problem and produces more and more synthetic latex products. PMID- 10818823 TI - AAOHN advisory. Getting results with psychiatric fitness for duty exams. PMID- 10818824 TI - Providing worksite health promotion through university-community partnerships. The South Carolina DOT project. AB - The occupational health nurse for the South Carolina Department of Transportation (SCDOT) collaborated with the Schools of Nursing within the state universities of South Carolina to coordinate individual health screenings for the employees of SCDOT. Personal Wellness Profiles (PWP) by Wellsource, Inc., were used to perform the health screenings and included family and personal histories, and assessment of blood pressure, vision, height, weight, total and high density lipoprotein cholesterol, and blood glucose levels. In addition, hepatitis and tetanus/diphtheria immunizations and influenza vaccines were provided. Each of the 48 county sites was visited twice during the semester by nursing faculty and nursing and public health students. The first visit was to collect assessment data for screening and provide immunizations. The second visit was to provide individualized analyzed data and health counseling. Of the 5,118 SCDOT employees, 3,141 were screened the first year and 2,315 were screened the second year. Of the original 3,141, only 1,549 elected to participate in the rescreening. Although the average population age was 41, 78.3% were rated at high coronary risk because of high blood pressure, high cholesterol and blood sugar levels, excessive weight and stress levels, and sedentary lifestyles. PMID- 10818825 TI - Work related assaults. The impact on victims. AB - This pilot project employed a case study design consistent with that of Rice to describe the impact of violence on individuals who incurred a work related assault in 1992. Ten randomly selected subjects were interviewed from a population of 429 individuals reporting a work related assault that resulted in a wage loss claim. Half of the subjects had received permanency ratings, thought to be a measure of injury severity. The study hypothesis, stating the impact of the assault (e.g., pain and suffering, decrease in functioning) years after a work related assault was associated with the severity of the injury (i.e., permanency rating), was not supported by the data. However, individuals' health and quality of life 4 years after the assault were affected significantly and resulted in job changes, chronic pain, changes in functional status, and depression. The new hypothesis resulting from this study is employer support and mental health intervention immediately after an assault may prevent employee job changes and decrease mental health sequelae. PMID- 10818826 TI - The effects of converting wheels on housekeeping carts in a large urban hotel. Program evaluation. AB - Occupational and environmental health nurses can identify the causes of injuries and use analytical skills to show how the prevention of a problem can save the company money and demonstrate a caring attitude from management. Nurses can expand their traditional roles to position themselves as both advocates for the employees and profit enhancers to management as demonstrated by this program evaluation project. Initial outcomes included improved employee morale and reduced soft tissue injuries in hotel housekeeping employees. Changes in the workplace made by a proactive occupational and environmental health nurse and a committed management can reap rewards beneficial for both the employer and the work force. PMID- 10818827 TI - Injury experience of temporary workers in a manufacturing setting. Factors that increase vulnerability. AB - An apparent two to three times higher injury frequency rate for temporary employees compared to permanent workers was identified in one manufacturing setting. Data were collected using demographic surveys (N = 20) and three focus group interviews (n = 13), with a convenience sample of temporary employees as well as four structured interviews with temporary agency owners and managers to explore factors which increase the vulnerability of temporary employees to workplace injuries. Several physical and psychological stressors were identified as well as a perception by the temporary employees that the reporting of work injuries could lead to loss of a temporary work assignment or the opportunity for permanent employment. In contrast, the agency owners and managers were concerned that reported injuries were not always legitimate. All of them actively used strategies to control their injury experience but faced the challenge of lacking day to day control of both the work environment and employee work behaviors. This preliminary study cannot be generalized to manufacturing worksites. Additional research is needed to document the injury experience of temporary employees, identify training needs, and design interventions. PMID- 10818828 TI - Office ergonomics. Measurements for success. AB - The successful implementation of an ergonomics program requires collecting data on worksite history repetitive motion injuries and assessing the corporate ergonomic needs. It is important to solicit management, department, and employee support. Program success depends on creating process, training program, skilled assessors, and an accountability method. A thorough understanding and application of neutral posture and the three seated chair positions is essential. Evaluation of the entire individual work-space is essential. Precise measurements of the individual, the office chair, and the work surfaces should be completed, and heights adjusted to meet individual needs. Functional, totally adjustable office chairs are a necessity. Vendor selection criteria and vendor contracts for chairs and office equipment assure consistent specifications and cost control. PMID- 10818829 TI - Electronic media storage. PMID- 10818830 TI - Adverse health effects among women living with heavy snorers. AB - Women living with heavy snorers were more frequently affected by symptoms of insomnia, morning headache, daytime sleepiness, and fatigue than women living with non-snorers. Questionnaire data were collected from 1,032 women 30 to 64 years of age residing in Dalarna county, in mid-Sweden. There were indications of a "dose-response relationship" between the conjectured sound exposure and reported symptoms, regardless of whether the female herself snored. Sleeping in separate bedrooms did not seem to give the women any alleviation. The results point to a possible contributory cause of disturbed sleep, morning headache, and daytime sleepiness among women living with a snoring spouse. The results also indicate that prevention and treatment of snoring are important issues for the couple as well as for the snorer. PMID- 10818831 TI - Treatment decision making in mature adults: gender differences. AB - Studies have shown that women are less likely to receive several specific medical interventions, including organ transplantation, cardiac diagnostic studies, and coronary artery bypass surgery. This study investigates the hypothesis that this inequity may be explained by gender differences in treatment decisions made by older adults. A self-report questionnaire using hypothetical situations, treatment choices, and influencing factors was designed based on literature review and interviews with key informants. The questionnaire was administered to 250 nonpatient adults over the age of 50 in urban and rural settings in Ontario, Canada. Results revealed no significant gender differences in hypothetical treatment decisions made by patients nor in the factors affecting those decisions. In the absence of evidence that patient choice accounts for gender differences in utilization rates, physicians need to carefully examine their assumptions about patient preference and gender in the provision of specific interventions to male and female patients. PMID- 10818832 TI - A feminist critique of research on women's work and health. AB - Research on women's work and health largely has failed to incorporate gender into the models of the processes through which work influences well-being. In this article, the research on women's work and health is critiqued from a feminist perspective; male-oriented and ethnocentric views on women's work are negated, and gender and socioeconomic issues are highlighted and included in the picture of women's work. Male-centered and ethnocentric views and assumptions on women's work are prevalent in the whole research process, and methodological limitations due to the distorted views are indicated. Based on the critique, some implications for future research on women's work and health are proposed. PMID- 10818833 TI - The use of natural seminal plasma in terminating intrauterine fetal death: a communication of the clinical trial. AB - Natural seminal plasma was used for the expulsion of dead fetuses in 153 cases of IUFD. The natural product was sterilized, diluted with saline, and injected extra amniotically into the intra-uterine space. The fetal expulsions in these cases were safe and were not accompanied by any complications. This is a novel method with simple procedures, low costs, and with a promising potential for further improvements and worldwide use. PMID- 10818834 TI - Factors influencing single mother's employment status. AB - Changes in the welfare system limit the length of time a person can receive welfare benefits, thus mandating employment for many current welfare recipients. Single mothers with young children who do not become employed will lose financial support for housing, food, clothing, and health care and place their own and their children's health and safety at risk. The purpose of this qualitative study was to explore women's experiences of being unemployed and to examine the barriers to employment perceived by single mothers who expressed a desire to be employed. Nine mothers were recruited from a larger sample of single mothers who had participated in a quantitative study about employment conducted 1 to 2 years earlier. Using focus group interviews, mothers were asked what it was like to be a single mother, and then what barriers to their employment they perceived. Two dimensions were identified from the mothers' statements. The first, a sense of obligation, included themes of "being there" for their own and their child's benefit and doing what it takes to optimize the child's growth and development. The second, negotiating the obstacles, referred to problems regarding child care, lack of involvement of the child's father and lack of support from relatives and friends for the mother's efforts toward securing employment. These findings have important implications for welfare reform, namely, that efforts aimed at moving nonemployed single mothers into the workforce will fail if these factors are not considered. PMID- 10818835 TI - Is home care a weak link? PMID- 10818836 TI - Caring for patients with latex allergies. AB - Natural Rubber Latex (NRL) is found in over 40,000 products that are used in healthcare settings as well as in the home. Providing care for the patient with a latex allergy requires understanding of how this allergy emerged, how it affects the patient, and what products are safe to use. PMID- 10818837 TI - Communicating with your supervisor: how to be heard. PMID- 10818838 TI - Nursing informatics: could this new specialty be for you? AB - The greatest challenge facing home care today is the need to provide the highest standards of care at a time when funds are cut and outcomes must be closely measured. From this challenge has emerged a specialty that integrates information science and computer technology with nursing science: nursing informatics. This article discusses the basics of the field and how home care nurses can best step into the role. PMID- 10818839 TI - Let stress make you--not break you. AB - Coping with stress is an important skill during these trying times. This article outlines some skills you can use to respond positively to stressful situations in the car, the office, and clients' homes. Coping strategies for dealing with stress in personal situations are also discussed. PMID- 10818840 TI - Integrating disease management and wound care critical pathways in home care. AB - This article discusses the need for an integration of the concepts of disease management and critical pathways as a foundation of a healthcare delivery system. The steps in the process for development, implementation, and evaluation of a wound care critical pathway are reviewed and variance classifications are defined. Co-pathways and algorithms are presented as methodologies for dealing with variances. A template of a wound care critical pathway that has been developed for use in the home care setting is included. PMID- 10818841 TI - How to assess and intervene in domestic violence situations. AB - The increasing incidence of domestic violence in America requires home care nurses to have a broad understanding of the problem. This article outlines the extent of domestic violent and lists specific ways in which nurses can integrate screening for domestic violence into routine assessments. Specific interventions and a comprehensive resource guide are provided. PMID- 10818842 TI - "Washington or bust". PMID- 10818843 TI - Snow go! PMID- 10818844 TI - Thanks to home health aides everywhere. PMID- 10818845 TI - Imagine this! Infinite uses of guided imagery in women's health. AB - Guided imagery, the use of focused concentration of formed mental images, provides the mechanism of an independent nursing intervention to facilitate mind and body healing. Nurse healers can channel clients to personal restorative potentials and independent health through this powerful and inexpensive tool. In a variety of outpatient, inpatient, chronic care, and home care settings, nurses can introduce this treatment modality early on and for lifelong use for any number of nursing diagnoses. Specific conceptualizations for women's health are presented here. By unleashing your own and your client's imagination, the endless possibilities of guided imagery applications and resulting self-empowerment become apparent. PMID- 10818846 TI - A holistic life view of human immunodeficiency virus-infected African American women. AB - Minority women represent the fastest growing segment of the population to acquire HIV/AIDS in the United States. Although African American women are a large proportion of this group, no published study has concentrated solely on a holistic view of the experiences of HIV-infected African American women. The primary purpose of this phenomenological pilot study was to describe the lives of five HIV-infected African American women. Audiotaped interviews were conducted and subsequently were examined, using Giorgi's steps of analysis. Twelve themes emerged from the data: violence, addiction, it couldn't happen to me, shock and denial, education, time, uncertainty, cycles, secretive nature of their lives, someone, survival, and children. It is clear, even from this small pilot study, that these women have complex experiences that must be better understood before effective health care interventions can be designed and implemented. PMID- 10818847 TI - The impact of premature menopause on women's experience of self. AB - Surgically or chemically induced premature menopause may have an impact on women's sense of self: the physical, psychological, social, and spiritual sense of self. This ethnographic research study explores the major factors that assaulted prematurely menopausal women's concept of self and the ways in which they attempted to readjust that concept. A unifying domain, evolution of self, and three constitutive themes emerged: change/loss, connectedness/disconnectedness, and transcendence/transformation. A holistic nurse-client relationship that transcends the hospital experience and extends into the community may help prematurely menopausal women to move toward a more synthesized sense of self and a greater degree of self-actualization. PMID- 10818848 TI - Journey to holism. AB - The journey to holism is the result of a grounded theory study of 18 members of the American Holistic Nurses' Association. Participants were prompted to "Tell me about holistic nursing" and replies revealed a seven-step process to becoming a holistic nurse. The process is an expansion of Van Gennep's three-stage theory of rites of passage, which involves separation, marginality/liminality, and reintegration. The expanded process begins with separation from mainstream nursing and concludes with reintegration into nursing as a holistic nurse. The marginality/liminality step shows five active steps: gathering information to quell the distress of the separation, applying that information to others, changing focus to self-care, applying the information to self, and increasing self-knowledge. This article describes the process and presents data to support the conclusion that the growth of a holistic nurse follows a recognizable course. PMID- 10818849 TI - Holistic interventions for acute pain episodes: an integrative review. AB - Pain is a universal experience that lends itself to nursing interventions based on a holistic approach. Pain is a dynamic process, and relaxation techniques are based on the recognition of the interaction between the physiological and psychological components of the human body. Although acute episodes of pain are found in all areas of nursing practice, limited research using relaxation techniques as supplements to pharmacologic intervention has been reported. This article reviews six research studies that have implications for holistic interventions in nursing practice. Theoretical bases, findings, and discussion of relevance for holistic nursing practice are presented as well as recommendations for future research in the area of relaxation technique use during episodes of acute pain. PMID- 10818850 TI - Early pregnancy prenatal diagnostic testing: risks associated with chorionic villus sampling and early amniocentesis and screening options. AB - Routine amniocentesis is an established test for the prenatal diagnosis of chromosome abnormalities and many single-gene conditions. Chorionic villus sampling and early amniocentesis, which are available in some centers, enable families to receive prenatal test results earlier in the pregnancy. There are concerns regarding the safety of early pregnancy testing because of increased risks for pregnancy loss and possible risks for birth defects. The use of first trimester testing has changed because of these concerns, and alternative methods of screening pregnancies at risk for a chromosome abnormality are being investigated. PMID- 10818851 TI - Carrier screening for cystic fibrosis: a perinatal perspective. AB - Cystic fibrosis (CF) is one of the most common autosomal recessive disorders in the United States today. One in 29 Americans, greater than 10 million people of Caucasian ethnicity, is a carrier for this genetic condition. Although it has been 10 years since the discovery of the gene for CF, carrier screening is not yet a standard of practice. The ultimate goal of carrier testing is to provide individuals at risk with information and guidance that will permit them to make informed and independent decisions. It can be anticipated that perinatal nurses will be part of the process of carrier screening of CF for prenatal patients. This article describes the nature of CF, the options for testing, and the basics of the molecular testing so nurses can be instrumental in the education process when this becomes standard practice. PMID- 10818852 TI - Noninvasive means of identifying fetuses with possible Down syndrome: a review. AB - Women who are 35 years or older are offered invasive prenatal testing because of the increased risk of chromosomal abnormalities, especially Down syndrome. In an attempt to increase the number of Down syndrome fetuses being detected and decrease the number of invasive procedures being performed on pregnancies not affected with a chromosome abnormality, both biochemical and ultrasound screening methods are being studied and are summarized in this article. The ultrasound markers reviewed include increased nuchal thickness, increased nuchal lucency, shortened femur, shortened humerus, pyelectasis, hypoplastic ears, echogenic intracardiac focus, hypoplasia of the fifth middle phalanx, and echogenic bowel. PMID- 10818853 TI - Genetic abortion: considerations for patient care. AB - Women who receive abnormal prenatal diagnosis results potentially face two emotionally difficult decisions. In this article, the first decision--whether or not to terminate the pregnancy--is presented with a discussion of the factors that may influence a women's choice. Women who choose to terminate the pregnancy face a second decision when more than one type of abortion procedure is available. Two second trimester abortion procedures--dilation and evacuation and labor induction--are compared and contrasted to delineate potential advantages and disadvantages of each. The decision-making process is examined, emphasizing the individual ways in which women may weigh this information to make a fully informed decision. In addition, a number of recommendations are offered to health care providers in the role of discussing options and supporting women in their choices. PMID- 10818854 TI - Chromosomal abnormalities: trisomy 18, trisomy 13, deletions, and microdeletions. AB - The birth of an infant with a chromosomal abnormality such as trisomy 18, 13, Wolf-Hirschhorn (4p-) syndrome, Cri-du-chat (5p-) syndrome, and the microdeletion syndromes creates a stressful and devastating experience for families. Many of these disorders have severe consequences encompassing major malformations and mental retardation. With increasing diagnostic accuracy, clinicians can now appreciate the wide variability and natural history of these disorders. Although many of these infants do not survive the neonatal period, others do. Neonatal nurses have the opportunity to assist families in these situations by providing anticipatory guidance and care. PMID- 10818855 TI - Prader Willi and Angelman syndromes: exemplars of genomic imprinting. AB - The molecular phenomenon genomic imprinting provides an explanation for why two clinically distinct syndromes share genetic etiologies. Increased understanding of genomic imprinting is affecting diagnostics. Use of improved diagnostic tests can enable early, syndrome-specific, and anticipatory interventions and consequently, improved quality of life; however, these tests are of little use unless clinicians are able to identify at-risk patients. Nurses knowledgeable about Prader Willi and Angelman syndromes and their associated genetic mechanisms can play a significant role in early identification, referral, and intervention of patients with these conditions. PMID- 10818856 TI - Neonatal nursing and the genetics revolution! AB - Discoveries from the Human Genome Project are having a significant impact on the way we look at genetic makeup, health, and disease. As a result of this new knowledge, changes will be made in how diseases are treated and how nurses work with patients and their families. The National Coalition for Health Professionals Education in Genetics was formed to strengthen baseline knowledge in genetics of nurses and other health professionals. For neonatal nurses, this is an extremely important area that will become more and more critical to care as genetics knowledge continues to rapidly expand. Neonatal nurses must be knowledgeable about genetics, genetic testing, and the use and possible misuse of genetic information. This article describes the impact of genetics on neonatal care. PMID- 10818857 TI - Implementation of a clinical pathway system in maternal newborn care: a comprehensive documentation system for outcomes management. AB - This article describes the design, implementation, and evaluation of an interdisciplinary clinical pathway system for maternal newborn care in a perinatal regional referral institution. Core issues in the design of this system are addressed to promote outcomes management and ongoing performance improvement. A discussion of the implementation follows, illustrating the lessons learned, changes made, and associated evaluation. This clinical pathway system has improved communication and collaboration among all disciplines, enhanced the discharge coordination process, and established protocols available to all members of the health care team. PMID- 10818859 TI - Nonimmune hydrops fetalis. AB - Nonimmune hydrops fetalis (NIHF) is a term that defines an edematous fetus that does not have erythroblastosis fetalis from isoimmunization. All of the other reasons that a fetus develops NIHF are included in this category. The causes of NIHF range from diseases to structural anomalies to genetic abnormalities. The mere presence of NIHF is a poor prognostic indicator, even with the etiology unknown. Some causes of NIHF are treatable with intrauterine therapies. These currently include cordocentesis to administer intrauterine transfusions or medications and fetal surgery. Prenatal testing to establish an etiology or follow the progress of the fetus is extensive, expensive, and stressful. The mortality rate remains high. Education, counseling, and support of the patient and family are the work of the prenatal period. Delivery room management of the infant with NIHF requires a resuscitation team with a specific plan of care to maximize chances of survival. PMID- 10818858 TI - Trauma in pregnancy. AB - Of all cases that come to hospital emergency rooms, the traumatized pregnant patient presents one of the most complicated dilemmas because nurses and doctors who commonly treat trauma victims rarely have comparable expertise in the management of pregnancy. Treatment and care of the traumatized pregnant patient are challenging because advanced pregnancy influences the pattern of trauma, alters laboratory values and clinical assessments, and changes hemodynamic parameters. Pregnancy may alter the usual trauma routines, and trauma may affect the outcome of the pregnancy. It is imperative that both the trauma team and perinatal team work collaboratively toward a common goal of resuscitating and stabilizing the pregnant woman without jeopardizing the fetus whenever possible. This article reviews alterations in anatomy and physiology that occur during pregnancy and discusses the impact of decision making by health care practitioners faced with the dilemma of managing trauma during pregnancy. PMID- 10818860 TI - Culturally competent nursing care during the perinatal period. AB - Effective nursing interventions require culturally competent nursing practice. Nurses can develop the skill sets needed to provide culturally competent care. These skills derive from nursing principles and practice that support respect for individual dignity and self-determination. Sweeping changes in health care delivery have shortened the length of client exposure to nursing care, especially in perinatal nursing. Moreover, changes in the United States population challenge the nurse's ability to respond appropriately to the expectations, values, and beliefs of many diverse cultural groups. Nursing theoretical frameworks provide a basis for cultural competence in practice. PMID- 10818861 TI - A clinical pathway system for the neonatal intensive care nursery. AB - Health care reform, managed care, and the current outcomes movement have generated a rapid acceleration toward the development and implementation of clinical pathways in the field of neonatology. This article describes the design, implementation, and evaluation of an interdisciplinary clinical pathway system for a neonatal intensive care nursery. This neonatal clinical pathway is a clinical tool that delineates practice guidelines for each discipline that provides care to a specific infant population. It has reduced variation in clinical process and thereby has been shown to improve the quality of infant care. When practice guidelines and documentation are linked to health and economic outcomes, they begin to significantly impact health care costs. PMID- 10818862 TI - Continuous insulin intravenous infusion therapy for VLBW infants. AB - Very low birth weight (VLBW) infants less than 1,000 g often experience hyperkalemia and hyperglycemia during the initial hospital course. Hyperkalemia has been noted in 44% to 50% of infants less than 800 g birth weight or less than 28 to 29 weeks' gestation. Hyperglycemia occurs 18 times more frequently in infants less than 1,000 g than in those weighing more than 2,000 g. Insulin has been used for VLBW infants less than 1,000 g to manage hyperkalemia, control hyperglycemia, and optimize parenteral nutrition. A protocol for using exogenous insulin therapy for VLBW infants is described. PMID- 10818863 TI - Infant handling in the NICU: does developmental care make a difference? An evaluative review of the literature. AB - Infant handling and disruptions in the neonatal intensive care unit are environmental stressors over which nurses have the most control. Two of the major goals of developmental care are individualizing care by decreasing infant disruptions and handling by caregivers, and modulating or attenuating infant responses to the care they receive. However, it has yet to be established to what extent these goals have been achieved. This article will provide a comparative review of selected literature to ascertain what effect, if any, the introduction of developmental care has had on infant handling or disruption in the neonatal intensive care unit. PMID- 10818864 TI - Schizophrenia: orthodoxy and heresies. A review of alternative possibilities. AB - Conflicts about the nature, causes and treatment of schizophrenia have never ceased. Recently, however, a particular set of beliefs ('the Orthodoxy') has become influential, and dominates political and managerial agenda for controlling clinical practice and educational initiatives. Theories and therapeutic strategies that differ from the particular biological, behavioural, cognitive and family management approaches favoured by orthodox clinicians have been given far less academic and clinical air-space. This review surveys some alternative ideas and practices ('the Heresies') and critically refers to some of the orthodox tenets such as the genetic and biological bases of schizophrenia, the disease model, 'family blaming' and drug treatment. Heresies briefly explored include schizophrenia as an evolutionary inevitability, creative sublimation of schizotypal tendencies and systems-theory-based family therapies. PMID- 10818865 TI - Towards understanding some complex borderline behaviours. AB - This article aims to explore and explain some possible antecedents to behaviours revealed by consumers in mental health settings who evoke strong and often negative responses in nurses and other health professionals. The contemporary exemplar par excellence is people who are diagnosed as having a borderline personality disorder. The paper discusses four different but overlapping domains that may contribute to understanding the difficulties many nurses face in relation to working with consumers whose behaviours are complex and distressing. Firstly, these begin with a brief discussion of psychiatric labelling and stereotyping associated with borderline personality disorder. Secondly, the connections between trauma in childhood and psychiatric vulnerability are explored, as nurses and other health professionals sometimes do not appreciate the debilitating and long-term consequences of the survival of childhood abuse or sexual assault. Thirdly, some unconscious defence mechanisms are outlined to partly explain behavioural interactions often displayed by people with a diagnosis of borderline personality disorder. Finally, behavioural concomitants of projection, splitting and projective identification are discussed: they include identity diffusion, self-mutilation and suicide attempts. All of these topics are explored to encourage nurses to support consumers to express pain in more constructive ways. PMID- 10818866 TI - Stress and burnout in forensic community mental health nurses: an investigation of its causes and effects. AB - The increasing development of forensic services has seen the numbers of Forensic Community Mental Health Nurses (FCMHNs) mushroom. Community mental health nursing (CMHNs) has been found to be a stressful occupation. FCMHNs have largely been ignored in research into occupational stress. The current study identified and surveyed all FCMHNs (n = 104) in England and Wales who were attached to NHS Medium Secure Units using measures of occupational stress. A response rate of 76.9% was achieved. Findings from this study showed that a substantial portion (44.3%) of FCMHNs were experiencing high burnout in relation to emotional exhaustion. A number (31.2%) of FCMHNs reached or exceeded the threshold for psychiatric caseness on one measure. Concerns of FCMHNs included 'not having facilities in the community to refer patients on to' and 'interruptions in the office'. Dealing with violent patients did not feature in the top 10 stressors, although dealing with suicidal patients did. As a group FCMHNs appear to feel supported by their managers and colleagues. PMID- 10818867 TI - Chronic mentally ill individuals reentering the community after hospitalization. Phase II: The urban experience. AB - The purpose of this qualitative study was to explore the experiences of persons reentering an urban community after hospitalization for mental illness. The sample consisted of five men and three women, aged early 30's to mid-50's, mostly diagnozed with schizophrenia, and who had had several hospitalizations within the previous year. Most returned to places where they had lived before, frequently to boarding homes for former psychiatric patients. Most lived alone. Each participant was interviewed shortly before discharge and one to two times in the community. The tape-recorded interviews were analyzed according to the Giorgi method. Three themes emerged from the data, related, respectively, to the hospital and its environs remaining a focus of the participants' lives, the added burden of social and financial conditions, and the presence of goals which nevertheless had barriers to their achievement. Contrasts are drawn with the findings of Phase I (Montgomery & Johnson 1998), in which there was found a much stronger sense of discharge marking a new beginning. The findings add to our knowledge about what it is like for chronic mentally ill individuals to live outside hospital, but also raise questions about the influence of particular diagnoses, community characteristics and changes in adjustment over time. PMID- 10818868 TI - General practitioners' perceptions of community psychiatric nurses in primary care. AB - The management of and responsibility for the care of people with mental health problems in the community is increasingly being assumed by general practitioners (GPs) and primary care personnel. As primary care groups (PCGs) evolve, so must their expertise in managing people with a wide range of mental health problems. It is expected that all mental health professionals will participate in this development, although it is likely that community psychiatric nurses (CPNs) will be the largest professional group involved, with a significant part to play in the shaping, management and delivery of mental health services. To date, there has been little research into how CPNs are perceived by other primary health care professionals. This study seeks to provide an insight into how GPs assess the contribution of CPNs in primary care. Overall, the results of the study suggest that GPs view CPNs favourably and consider that they have an important role to play. Greater involvement in primary care raises issues about the education and preparation of CPNs, their professional development and supervision needs. PMID- 10818869 TI - Suicidal patients and special observation. AB - Special observation is a nursing practice utilized in in-patient psychiatric facilities for patients who are suicidal. Special observation is carried out by an allocated registered nurse remaining continuously at arms length from, or within sight of (as specified), the patient; or a registered nurse observing the patient within a 30 min interval. In this exploratory study, we investigated the role of the registered nurse when caring for patients on special observation. Semistructured interviews were conducted with 10 randomly selected registered nurses from 4 acute psychiatric wards. The 9 themes that emerged were: 1) Safety; 2) Therapeutic relationships; 3) Supporting patients and carers; 4) Consequences of special observation for nurses; 5) Continuity of care concerns; 6) Peer support; 7) Suicide indicators; 8) Responsibilities and rights: nurses and patients; and 9) Nurses, doctors and the hospital hierarchy. Recommendations for alleviation of the concerns about the practice of special observation are outlined and discussed. PMID- 10818870 TI - 'Something always comes up': nurse-patient interaction in an acute psychiatric setting. AB - In this study, 10 nurses and 10 patients were interviewed to explore factors influencing nurse-patient interactions in an acute psychiatric inpatient facility. The six themes that emerged from the nursing interviews were; environment, something always comes up, nurses' attributes, patient factors, instrumental support and focus of nursing. The four themes from the patient interviews were; nurses' attributes, role perceptions, clinical care, and time. These findings have implications for clinical practice, the nurses' role and nursing education. PMID- 10818871 TI - Who wants to be a psychiatric nurse? Novice student nurses' interest in psychiatric nursing. AB - Available research findings suggest that nursing students do not regard psychiatric nursing as a desirable future career option. This paper addresses the results of a research project conducted by the author. This research addresses the relative popularity of psychiatric nursing in comparison to other nursing specialties, within Victoria, Australia. The research was conducted by use of a questionnaire in which commencing undergraduate nursing students were asked to rank nine areas of nursing specialty in order of preference and provide some explanation for their choices. The results indicate that psychiatric nursing emerges as the second least popular career choice for student nurses at this stage of their education. PMID- 10818872 TI - Involving service users and carers. PMID- 10818873 TI - Defining severe or enduring mental illness: by principle or prescription? PMID- 10818874 TI - On becoming an author: writing for the Journal of School Nursing. PMID- 10818875 TI - Developing a drug awareness program in an international school. Cross-cultural issues. AB - In 1996, as an overseas school nurse/health educator, the author designed a health and personal development curriculum for an international school in Bandung, Indonesia, where 220 children from 26 different countries were enrolled. Part of the health curriculum included a drug awareness program for students from kindergarten through high school. Many parents, students, administrators, and faculty had never been involved in such a program before; therefore, obtaining acceptance from these groups was a first step in designing the drug awareness program. Because there have been no school or government anti-drug groups to promote drug prevention, this program was the first of its kind in the international community of Bandung. A review of the literature guided the choice of intervention strategies built into the program. Information also was collected about the major risk and protective factors that are known to be associated with an increased risk of drug use in the international community. Existing models of drug use prevention were used in designing the program. As in most prevention programs, drug use was viewed as a deficit in coping or self-regulation skills. The drug awareness program was developed with emphasis on peer, school, and community factors potentially influencing the tobacco, alcohol, and other drug use of the adolescent students. PMID- 10818876 TI - A state association surveys school nurses to identify current issues and role characteristics. AB - A state association of school nurses conducted a statewide survey, collaborating with university nursing faculty to secure necessary resources and research expertise. A questionnaire, including demographic, employment, and practice characteristics, was mailed to 340 school nurses in Michigan. The response rate was 49%. Findings demonstrated the complex, multiple roles and responsibilities of school nurses. Two strengths identified were memberships in professional organizations and employer support for continuing education. Issues identified included educational preparation below the recommended baccalaureate level, lack of differentiation in practice based on education, and certification through non competency-based processes. Isolation and supervision/evaluation by non-nurses were pervasive themes. Findings are relevant for school nurses across the country. Results have been used both locally and at the state level in employer negotiations, with legislators for educational purposes, and in policy development. Strategies for addressing the identified issues are explored, including those facilitated through an association-university collaboration. PMID- 10818877 TI - An ethical dilemma in school nursing. AB - School nurses must consider the ethical principles that guide everyday practice. Autonomy, beneficence, justice, and nonmaleficence are the ethical principles most often confronted in the school setting. When beneficent care-giving begins to conflict with the family's decision-making autonomy, paternalism, a form of beneficence, affects the family's autonomy. This creates an ethical dilemma for the school nurse who guides his or her practice by ethical principles but who also must decide when it is appropriate to refer a child or family to a medical provider for further evaluation. A case study is presented to illustrate a specific ethical dilemma. The ethical dilemma is described using a model that examines external factors, professional responsibilities, and possible courses of action. The discussion includes cultural considerations and barriers pertinent to the case example. PMID- 10818878 TI - Attitudes about menstruation among fifth-, sixth-, and seventh-grade pre- and post-menarcheal girls. AB - A descriptive study of 106 fifth-, sixth-, and seventh-grade girls was conducted to determine their attitudes toward menarche. Attitudes of affirmation and worry were examined based on grade level, menarcheal status, and various other variables, including whether the participants had talked with their mother or a close friend, or seen a video on menstruation. Worry scores increased independently of menarcheal status from the fifth to sixth grade. From the sixth to seventh grade, there was no change in the level of worry of post-menarcheal girls, but there was a significant decrease in worry for pre-menarcheal girls. Perhaps these girls sought reassurance from a supportive person that their late menses was not abnormal. If so, this reassurance may have accounted for their decrease in worry scores. There was a decrease in worry in those girls who had sisters. Also, girls who had talked with someone about menstruation had higher affirmation scores than those who had not. Nursing implications include the need for school nurses to provide ongoing education for young females from fifth through seventh grades, and to encourage them to communicate with their support systems. PMID- 10818879 TI - Online continuing education. AB - Nurses looking for learning opportunities often have to travel many miles to find content that applies to their specialty area. The World Wide Web has continuing education offerings with content relevant to school nurse practice and the care of children in the community. The Web allows the convenience of taking courses at the student's choice of time, place, and pace. PMID- 10818880 TI - Creating a living document. The National Association of School Nurses Strategic Plan 1998-2001. AB - In all organizations, periodic assessments are prudent investments in the future. For the National Association of School Nurses (NASN), the dramatic changes in health care and their influence on school nurses, the shifting needs of schoolchildren, the ending of the time frame of NASN's previous strategic plan, and the need to better position the Association to identify and provide for the current and future needs of the membership, led NASN leaders and management staff to initiate an assessment of member needs. This assessment is the prelude to the development of the 1998-2001 Strategic Plan. The way in which it was accomplished -and the impact on school nurses--is described in the first of a series of three articles. Two future Nursing Practice Management section articles will discuss the Association's mission statement and suggest strategic planning for all school health programs as well as the implementation of the seven goals in the 1998-2001 National Association of School Nurses Strategic Plan. PMID- 10818881 TI - Use of lumbar cerebrospinal fluid drainage in thoracoabdominal aortic aneurysm repairs. AB - Thoracoabdominal aortic aneurysm repairs present many challenges, and the complication of paraplegia remains a concern for both the surgeon and the nurse caring for the patient in the postoperative period. Paraplegia can occur secondary to spinal cord ischemia from prolonged aortic clamping during the repair of the descending thoracic aorta. Paraplegia is a devastating complication for the patient and family. Multiple adjunct techniques have been instituted to prevent reduced spinal cord perfusion during and after the operation, including the use of shunts and cardiopulmonary bypass, femoral artery-femoral vein bypass, left atrial-femoral artery bypass, and selective revascularization of the dominant intercostal artery. Other methods, such as somatosensory evoked potential monitoring during the operation and regional spinal hypothermia techniques, have not reduced the incidence of paraplegia. Improved outcomes have been seen with the use of methods to reduce cerebrospinal fluid (CSF) pressure. One such method is the use of external CSF drainage during the operation, followed by use of a lumbar drain system for as long as 72 hours after the operation. This system setup uses a transducer to monitor CSF pressure and a drip chamber to drain CSF to maintain a normal pressure. This article describes thoracoabdominal aneurysms, surgical techniques to repair the aneurysm, and the use of external CSF drainage and related nursing care measures. PMID- 10818882 TI - Ultrasonographic diagnosis and guided compression repair of femoral artery pseudoaneurysm: an update for the vascular nurse. AB - Rapid advances in technology have led to the use of radiographic technology for therapeutic interventions. This article is an all-inclusive procedure guide for the imaging nurse as well as the bedside nurse caring for the patient before or after the procedure. Key aspects of the nurse's functions are educating the patients to gain their cooperation and improve their understanding of the procedure and post-procedure expectations, physiologic monitoring, and administering sedation and analgesia. Ultrasonographic guided compression repair is a safe, cost-effective first approach to treatment for many femoral artery pseudoaneurysms and does not affect possible surgical intervention if the ultrasonographic guided compression repair attempt fails. PMID- 10818883 TI - A new approach to an old and vexing problem: subfascial endoscopic perforator surgery. AB - Chronic venous insufficiency with venous hypertension causes leukocyte trapping, lipodermatosclerosis, and finally, skin ulceration involving the lower extremity. Perforator vein incompetence has been identified as an important contributing factor to ulceration when abnormally elevated pressure is transmitted to areas of affected skin, usually at the ankle medially. Surgical techniques for ligation of incompetent communication veins were first popularized by Linton and Dodd from 1940 to 1950. Early techniques used extensive longitudinal incisions for subfascial ligation through indurated skin. These procedures were plagued with wound complications: delayed healing, skin necrosis, and infection. Techniques continued to evolve that used minimally invasive incisions and avoided zones of affected skin. With the availability of endoscopic, fiberoptic, and laparoscopic advances in surgery, instrumentation has been developed for minimally invasive endoscopic approach to accomplish subfascia endoscopic perforator surgery (SEPS) under direct vision. SEPS is now used alone and in combination with other venous interventions to reduce transmission of venous hypertension to affected skin areas. The SEPS procedure, its indications, and the history of surgical treatment of perforator vein incompetence are discussed. Unique problems related to short hospital stays and postoperative care are outlined. This review will help the vascular nurse understand the rationale and techniques of SEPS. This comprehension will enable provision of accurate information to the patient and a knowledge-based plan of care. PMID- 10818884 TI - Extraanatomic bypass surgery. AB - The term "extraanatomic bypass" applies to grafts that pass through a different anatomic pathway than do the natural blood vessels they replace. These grafts are devised to circumvent complex problems when conventional vascular procedures are not possible or are too hazardous to perform. There are various indications, advantages, and disadvantages to each of these grafts. Extraanatomic bypass grafts are an accepted technique with known patency rates. Quality nursing care plays a significant role in successful patient outcome. PMID- 10818885 TI - Relieving intermittent claudication: a nursing approach. AB - Peripheral arterial disease affects at least 10% of adults older than 70 years. Risk factors such as diabetes, hypertension, hyperlipidemia, history of smoking, and genetics increase the incidence of the disease. Intermittent claudication, experienced as calf pain or cramping, is the primary symptom in patients with lower-extremity peripheral arterial disease. Patients with claudication are unable to walk even moderate distances. As a result, they often lead lives that are profoundly restricted. Medical therapeutic options available for patients with intermittent claudication are limited to a small number of medications and walking exercise rehabilitation. Walking exercise training can significantly increase ability and decrease calf discomfort for many patients. Nurses can have a major impact on improving the quality of life of patients with claudication, not only by seeking referrals to established institutional walking exercise programs, but also by helping patients in the community develop a personalized walking program. In this article, a nursing plan of care including short-term and long-term goals is addressed. A case study will illustrate the effectiveness and improved quality of life that an individualized program of walking exercise had for one community-based client. PMID- 10818886 TI - Effects of vascular surgery on the elderly vascular patient. AB - The majority of vascular patients are elderly and present a unique set of problems after an operation. Age plays a major role in their recovery, but the greatest challenge is their preexisting medical problems. The changes that occur with aging in the following body systems will be discussed: cardiac, pulmonary, renal, gastrointestinal, genitourinary, and central nervous system. Special concerns related to pain management, risk of delirium, and wound healing present continuing nursing challenges that require close observation after surgery. PMID- 10818887 TI - Compression wraps for venous ulcer healing: a review. AB - Compression to the lower extremities is used to increase healing of venous stasis ulcers by improving the blood supply and reducing edema and distension. Compression wraps are available in elastic or non-elastic and in single to multilayer systems requiring varying types of application and exerting different levels of compression. Elimination of edema is so basic and important to venous ulcer healing that the most effective level of compression should be used. Controversy exists regarding the most effective sub-bandage pressure (ranging from 20 mm Hg to 45 mm Hg) for timely healing. Because of differences in compression wraps, selection of the most effective and efficient wrap can be difficult. The purpose of this integrated review was to determine healing rates of venous ulcers with various wraps. The studies reviewed provide reasonable evidence that venous ulcers can be healed with the use of compression wraps and that various wraps are effective when used with correct assessment, application, and fit by the caregiver, along with compliance and mobility of the patient. PMID- 10818888 TI - Limb loss: alterations in body image. AB - An estimated 10% of persons older than 70 years have peripheral vascular disease. Peripheral vascular disease includes venous and arterial medical conditions. Peripheral arterial disease (PAD) is a major concern in assessment of patients for loss of limb. With the ever increasing numbers of senior citizens in the United States today, a major public health concern is the management of chronic health problems. Many persons who have progressive PAD as a result of circulatory changes eventually are forced to have a limb amputated. This amputation triggers the realization of the loss of the limb, which directly influences body image. Therefore, management of the effect of limb loss on body image becomes a major nursing consideration in health promotion/health maintenance endeavors by health care professionals. This article explores the concepts of limb loss and body image. The relationship between PAD and limb loss is established. The conceptual link between limb loss and body image alteration is described, with emphasis on the importance of nursing interventions to manage body image alterations. Pathophysiologic conditions leading to PAD and limb loss are reviewed. Nursing implications for management of body image alterations with limb loss are summarized. PMID- 10818889 TI - Internal carotid artery occlusion. PMID- 10818890 TI - Use of a silicone net dressing in severe mycosis fungoides. PMID- 10818891 TI - The role of a hydrofibre dressing in squamous cell carcinoma of the anal canal. PMID- 10818892 TI - Measurement of interface pressures in the evaluation of operating theatre mattresses. AB - This study compared the sacral interface pressures obtained in four mattresses (all supplied by the same manufacturer), in two positions adopted for surgical procedures, to determine the similarities and differences between them. The mattresses were all variations of the standard product used in operating departments. The null hypotheses were that there are no differences in interface pressures between the mattresses or between the two surgical positions, and that measurements are not influenced by subject body mass index. The measurement technology was also tested for reliability and suitability for intra-operative use. Sacral interface pressures of 25 healthy volunteers were recorded using each of the four mattresses (order individually randomised) in both the supine and the Lloyd Davies positions. A pressure-sensitive mat was used and one-way analysis of variance was the main statistical approach. Statistical significance was determined by p-values of < or = 0.05. One mattress recorded significantly lower interface pressures than the others and all pressures were influenced by body mass index. Surgical position was a significant factor, measurements in the Lloyd Davies position being 9.5-14.2% higher than in the supine position. The pressure measurement technique has proved to be reliable and easy to calibrate, position and use. Indications are that it can be used safely and effectively in operating departments. These results have implications for researchers and clinicians. PMID- 10818893 TI - A comparison of sub-bandage pressures produced with two multi-layer bandaging systems. AB - Fourteen nurses with experience in the use of high-compression bandaging were asked to bandage the same limb with two different bandaging systems: the 'Charing Cross' four-layer regimen (System A), and a modified system incorporating two new bandages (System B). A sub-bandage pressure monitor was used to quantify the efficacy of the resulting bandaging for the two systems. The results indicated that there was no significant difference between site sub-bandage pressures achieved using the two bandaging systems. Sub-bandage pressure profiles, however, fluctuated with patient posture, the best profiles being obtained with System B and the patient in the sitting position. PMID- 10818894 TI - Management of wound sinuses. PMID- 10818896 TI - Allergic reactions to dressings. PMID- 10818895 TI - Comparison of two dressings in the management of partial-thickness donor sites. AB - This study evaluated and compared the performance of an adhesive hydro-cellular dressing with that of a paraffin gauze dressing in the treatment of partial thickness skin-graft donor site wounds. Fifty patients were included in the study, each acting as his/her own control. Donor site area ranged from 20 cm2 to 71 cm2; half the area of each patient's donor site was treated with the trial dressing, the other half with paraffin gauze. Outcome measures assessed were: time to complete epithelialisation; ease of dressing removal; pain on removal; and appearance of the wound bed. The trial dressing demonstrated a significantly faster healing time (p < 10(-6)) and enhanced patient comfort. PMID- 10818897 TI - Foot impulse technology in arteriovenous disease. PMID- 10818898 TI - Motivation and compliance in wound management. PMID- 10818899 TI - The influence of nutrition and sepsis upon wound healing. PMID- 10818900 TI - The development of an in-house leg ulcer management course. PMID- 10818901 TI - The Steven Johnson syndrome. A case study. AB - Steven Johnson's Syndrome is a serious systemic disorder in which there are vesicobullous lesions involving the skin and mucous membranes. It can result as an immune response to an antigen or as a drug reaction. Most often it is considered as an allergic reaction. It is a self-limiting condition which responds to immediate management or may result in fluid loss, sepsis and death. PMID- 10818902 TI - Time to move towards a private nurse practitioner. PMID- 10818903 TI - Tasks and challenges in nursing. A note in perspective. PMID- 10818904 TI - High risk of violence against nurses. PMID- 10818905 TI - Is clinical audit under threat? PMID- 10818906 TI - The art of delegation. PMID- 10818907 TI - No workaholics need apply. PMID- 10818908 TI - Return ticket. PMID- 10818909 TI - Achieving uniform standards. PMID- 10818910 TI - Quality product. PMID- 10818911 TI - Patients know best. PMID- 10818912 TI - Dame Yvonne Moores. Interview by Tom Keighley. PMID- 10818913 TI - Career nurse, or nursing as a career: Part 2. PMID- 10818914 TI - Deliberate self harm in medium security. PMID- 10818915 TI - Leading from the front. PMID- 10818916 TI - A defining time in health care. PMID- 10818917 TI - Questions & answers from the JCAHO. What new standards are in store for 2000? AB - Preparing for 2000? So is the Joint Commission. Learn about changed standards that will impact nurse leaders in the Comprehensive Accreditation Manuals for Ambulatory Care, Behavioral Health Care, Health Care Networks, Home Care, Hospitals, and Long-term Care. PMID- 10818918 TI - File a complaint, face retaliation? AB - Sometimes it's okay to fire an employee after she files a complaint. Learn how to recognize unacceptable measures to avoid or defend lawsuits. PMID- 10818919 TI - R&D in the mergers and acquisitions age. AB - Learn about an emerging shift in the way research and development (R&D) is funded and the implications of that shift for health care information technology, product innovation, and delivery. PMID- 10818920 TI - How to mobilize an organization with commitment, not cash. AB - Leaders at an 852-bed community hospital planned an elaborate redesign that would reorganize units, revamp rooms, and update the exterior. With all the planning in place, the organization took a financial decline. Learn how nurse leaders and staff pulled together to accomplish their goals, save money, and serve patients better. PMID- 10818921 TI - Help the health care team release its hold on restraint. AB - As part of a three-site cooperative physical restraint reduction program in acute care hospitals, a multidisciplinary team created a survey instrument to measure staff's knowledge, unit beliefs about practice patterns, ethical concerns, and more. PMID- 10818922 TI - Poised for next-century politics? AB - From management hierarchies, to leveraging "who you know," to shared governance, organizational politics have changed over the years. Best bets for the future: Be a good leader, don't get hung up on titles, and value your employees. PMID- 10818923 TI - How to lead departments outside your clinical competence. AB - Leading departments outside your clinical competence takes a special set of skills. Follow these seven approaches and enhance your transferable leadership skills to succeed. PMID- 10818924 TI - Acute thoracic aortic dissection: can your nurses defuse a time bomb? AB - Sometimes, every second counts. Help your staff learn to differentiate between acute thoracic aortic dissection and myocardial infarction, how to confirm a dissection, and how to care for aortic dissection patients. PMID- 10818925 TI - Teach nurses effective ways to deal with inadequate staffing. PMID- 10818926 TI - Occupational exposure. Nurses who test HIV-positive need your support in their time of crisis. PMID- 10818927 TI - Vascular access devices: lines to live by. AB - Vascular access devices take many forms--all designed to maximize treatment options, reduce risks to patients and clinicians, and deliver optimal patient outcomes. PMID- 10818928 TI - Ask AONE's experts ... about how to reduce overtime and use of per diem staff. AB - Nurse leaders reduce overtime and use of supplementary staff by establishing resource pools, allocating staff based on nursing hours instead of full-time equivalents, and more. PMID- 10818930 TI - Start a disease state management program. PMID- 10818929 TI - Questions & answers from the JCAHO. When to log refrigerator temps. AB - The Joint Commission has standards for appropriate storage of medication, as well as food and nutritional products. Learn what the standards say about your staff watching the "fridge." PMID- 10818931 TI - What should you expect from your attorney? AB - If your're ever involved in litigation, you'll need legal support. Use a client bill of rights to build a good relationship with your attorney. PMID- 10818932 TI - Can Medicare beneficiaries pay outpatient drug bills? AB - This article updates nurse managers on the social and political aspects of how Medicare beneficiaries pay for outpatient drug bills. PMID- 10818933 TI - Intranet systems offer fast access to policies and procedures. AB - Still using paper policy and procedure manuals? Here's how one hospital staff switched from paging through bound photocopies to performing quick searches in online manuals. PMID- 10818934 TI - Enrich your performance coaching techniques. AB - Learn to read your employees' perspectives and recognize the personal traits that may impede professional performance. Then start a coaching dialogue to set goals together and reach them. PMID- 10818935 TI - Nursing leaders predict top trends for 2000. AB - Nursing Management's Editorial Advisory Board predicts the top 10 trends that will impact nurse leaders in 2000. Use them to lead in the new millennium. PMID- 10818936 TI - Plan faster, healthier recovery after orthopedic surgery. AB - Two hospitals--one community and one urban--use clinical pathways to streamline patient care. They lower the average length of stay by 41% and 30%, respectively, and shrink clinical and surgical complications. PMID- 10818937 TI - Do you help staff rise to the fall-prevention challenge? AB - Falls aren't normal occurrences for the elderly population. Nurses can prevent falls by assessing patients for risk and taking steps to prevent falls and minimize their effects. PMID- 10818938 TI - Win-win mentoring. AB - Learn strategies to build and enhance mentor-protege relationships, expand your experience, and help new nurses develop. PMID- 10818939 TI - Desperate strategies for last-minute staffing. AB - When a nurse calls in sick, don't fall apart--leap into action! The author humorously tells how to solve desperate staffing moments. PMID- 10818940 TI - Nursing takes a dual perspective on the future. AB - Nurses in service and academia joined to form the Nursing Resource Coalition in Southern California. The group works together to tackle common issues, such as shared resources, nursing's future, and measurement of vital indicators in both arenas. PMID- 10818941 TI - How to promote pulmonary health with kinetic therapy. AB - Kinetic Therapy can improve outcomes for immobilized patients, but the treatment can be costly. Using a protocol designed by critical care nurses, nurses in your units can provide cost-effective Kinetic Therapy. PMID- 10818942 TI - Restricted versus open ICUs. AB - A satisfaction study investigates whether a more liberal ICU visitation policy satisfactorily meets visitors' and nurses' needs and expectations. PMID- 10818943 TI - How do you rate your team's teamwork? PMID- 10818944 TI - Travel and registry nurses help fill staffing gaps. PMID- 10818945 TI - For the patient's sake, communicate! PMID- 10818946 TI - POC: testing on the move. AB - Learn the applications of point-of-care testing devices and how to decide if they're right for your department. PMID- 10818947 TI - Ask AONE's experts ... about how to implement a palliative-care program. AB - Learn what steps you should take to implement a palliative-care program that capitalizes on your institution's strengths. PMID- 10818949 TI - Don't forget the Journal Club! PMID- 10818948 TI - Time for action on poor weaning diets. PMID- 10818950 TI - Sleeping "problems" in babies and children: 1. When babies are wakeful, who has the sleeping problem? PMID- 10818951 TI - Meningitis: a new vaccine but the vigilance must go on. PMID- 10818952 TI - Kicking the habit: aids to giving up smoking. PMID- 10818953 TI - Why eat breakfast? PMID- 10818954 TI - Denying treatment to Down's syndrome children. PMID- 10818956 TI - School nursing in Moscow. PMID- 10818955 TI - Anorexia: the warning signs. PMID- 10818957 TI - The state of our vital resources. PMID- 10818958 TI - Bioterrorism: an overview. AB - How real is the threat of bioterrorism? Experts may disagree on the likelihood of use, but the possibility cannot be totally dismissed. Complacent ignorance of a low-probability, high-cost risk is dangerous and can result in devastating global consequences. PMID- 10818959 TI - Smart training: the US Army's pilot project for combat trauma surgical training. AB - Military medical personnel have an important wartime mission. To enhance combat trauma readiness and increase training opportunities, the military is experimenting with providing this additional trauma training in a civilian level 1 trauma center. PMID- 10818960 TI - Information systems disaster planning in the patient care environment. AB - This article focuses on preparedness planning for the nurse manager in the event of a network failure of a hospital clinical information system. The evolution of health care information systems is discussed. A 4-step preparation model is discussed to aid the nurse manager in preparing for network failure. PMID- 10818961 TI - Violence in the workplace: violence prevention strategies in the operating room. AB - Violence in the workplace has escalated, becoming a growing occupational hazard for health care workers today. Training and education strategies to de-escalate and prevent the rise in violent assaults in the health care setting are needed. A violence prevention program was developed at William Beaumont Army Medical Center to educate the operating room (OR) staff about prevention and intervention strategies to de-escalate or prevent violent assaults in their workplace. PMID- 10818962 TI - Malignant hyperthermia: a case study. AB - Malignant hyperthermia continues to be a life-threatening emergency that can occur without warning. With early discharge, this crisis may even occur at home. Perioperative, anesthesia, and Post-Anesthesia Care Unit (PACU) nursing staff need to be educated in the signs, symptoms, treatment, and care of a malignant hyperthermia patient. This is a US government work. There are no restrictions on its use. PMID- 10818963 TI - Transfusion reactions due to Yersinia enterocolitica. AB - The transfusion of blood products has become increasingly safe over the last few years, but the procedure is not without risk. One rare, but serious, complication is sepsis related to the transfusion of blood contaminated with bacteria. This article discusses the risk, treatment, and prevention of this potential complication of transfusions. PMID- 10818964 TI - The early history of orthopaedic surgery in Philadelphia. 1976. PMID- 10818965 TI - Musculoskeletal surgery in eighteenth century America. AB - Surgeons in America in the eighteenth century treated wounds, fractures, dislocations, and gunshot injuries of the skeletal structures and also contended with osteomyelitis and tuberculosis of bones and joints. In meeting this challenge, they proved innovative, adaptive and courageous, and developed principles of surgery, some of which still are in use today. PMID- 10818966 TI - The evolution of orthopaedic surgeons from bone and joint surgery at the University of Pennsylvania. AB - When the first medical school in the North American colonies was founded in 1765 at the Philadelphia College (University of Pennsylvania) there were only two branches of medicine physic (medicine) and surgery. Surgeons such as Philip Syng Physick, and his successors William Gibson, Henry Hollingsworth Smith, and D. Hayes Agnew, in addition to performing general surgery, treated patients with orthopaedic, ophthalmologic, and nuerosurgical problems. Treatment of patients with orthopaedic problems by surgeons continued at the University of Pennsylvania until DeForest Willard founded the Department of Orthopaedics in 1889. In the interval between 1805, when Physick was appointed the first professor of surgery at the University of Pennsylvania, and 1889 many ingenious instruments, splints, and operative procedures for treating patients with orthopaedic problems were developed. The author will describe some of the accomplishments of these pioneers. PMID- 10818967 TI - History and development of trauma care in the United States. AB - Until recently the development of systems for trauma care in the United States has been inextricably linked to wars. During the Revolutionary War trauma care was based on European trauma principles particularly those espoused by the Hunter brothers. Surgical procedures were limited mostly to soft tissue injuries and amputations. The American Civil War was remarkable because of the contributions that were made to the development of systems for trauma care. The shear magnitude of casualties required extensive infrastructure to support the surgeons at the battlefield and to care for the wounded. For the first time in an armed conflict, anaesthetics were used on a routine basis. Despite these major contributions, hospital gangrene was a terrible problem and was the cause of many mortalities. World War I and World War II were noteworthy because of the contributions made by surgeons in the use of blood. One of the major lessons of World War II was the reemphasis of how frequently lessons have to be relearned regarding the treatment and care of wounds. Between the Korean Conflict and the Vietnam War the discovery was made of the tremendous fluid shifts into the cell after severe hemorrhagic shock. As a consequence, the treatment of patients with shock was altered during the Vietnam Conflict, which resulted in better outcomes and less renal failure. The first trauma centers for civilians were started in the United States in 1966. Since 1988 the number of states with mature trauma systems has expanded from two to 35. During the same period, many studies have documented the efficacy of trauma systems in reducing unnecessary mortality and disability. PMID- 10818968 TI - Boston's contributions to the development of orthopaedics in the United States. A brief history. AB - Since the early part of the nineteenth century, physicians from Boston have had a major impact on orthopaedics in America. Initially, the general surgeons such as John Ball Brown and his son, Buckminster had an impact on orthopaedics, but contributors such as Henry Bigelow and Charles Scudder added greatly to the knowledge and capacity for care. The first orthopaedic ward, Ward I, was located at the Massachusetts General Hospital and began the new era. Surgical treatment by orthopaedists began in the twentieth century with such contributors as Philip Wilson, EA Codman, Robert B. Osgood, Joel Goldthwait, Elliot Brackett, Robert Lovett, and Edward Bradford. These physicians not only treated patients at the Children's, Tufts University, Boston City, the Beth Israel, and the Massachusetts General Hospitals, but assumed the academic responsibilities of a major educational center. More recently the contributors to orthopaedics have included Marius Smith-Petersen, Otto Aufranc, Henry Banks, Edward Cave, Carter Rowe, Joseph Barr, and others who have created a spectacular program for education, research, and clinical care. PMID- 10818969 TI - The early history of arthroplasty in the United States. AB - Arthroplasty is defined in the broadest sense as a reconstructive procedure that alters the structure or function of a joint. The first recorded procedures done in the United States in the early nineteenth century and the introduction of modern total joint replacement in the 1970s will be discussed. Although major surgical procedures occasionally were performed in the early 1800s, it was not until the introduction of general anesthesia and antiseptic techniques during the latter half of the nineteenth century that the field of surgery could be developed. Procedures involving the major joints of the upper and lower extremities are described. These procedures include resection and interposition arthroplasties, joint debridement, procedures done to correct complications of hip fractures and developmental dysplasia of the hip, cup arthroplasties, endoprosthetic replacement, hinge arthroplasties, resurfacing procedures, and early total joint replacement. PMID- 10818970 TI - Hospital for special surgery. A brief review of its development and current position. AB - On May 1, 1999, the Hospital for Special Surgery was 136 years old. To present a history that does adequate justice to the many people, who have been or still are involved in the making of it, is an impossible task. Nevertheless, this document is important because the hospital, first under the name of Ruptured and Crippled and then under that of Special Surgery has played such an important role in the development of orthopaedics and rheumatology in America during the past century. The reader must forgive an orthopaedic bias to this account and also realize the limitation of space that makes it at best fragmentary and incomplete. The account begins with a description of the hospital's current situation, physical layout and governance. A terse history of the hospital's origin and subsequent development follows, which includes a more specific description of the growth of its orthopaedic surgical services. Finally, separate records of the 112-year-old Postgraduate Orthopaedic Educational Program and 44-year-old Research Division are presented. PMID- 10818971 TI - The career and orthopaedic injuries of Joshua L. Chamberlain. The hero of Little Roundtop. AB - Joshua L. Chamberlain, a professor of religion and oratory at Bowdoin College, Brunswick, Maine was commissioned Lieutenant Colonel of the 20th Maine Volunteer Regiment in August 1862. He commanded the 20th Maine Volunteer Regiment in the historic defense of Little Roundtop against overwhelming odds and circumstances during the pivotal battle of Gettysburg. In June 1864 while leading a charge at the battle of Petersburg, he sustained what was thought to be a mortal wound when he was struck by a minie ball that entered just below and anterior to his right greater trochanter, injuring vessels, the urinary bladder and urethra, and fracturing his pelvis. Surgery was done in the field hospital, and after 5 months of recuperation, Chamberlain returned to lead his troops in combat. After the war he was elected Governor of Maine for four terms and subsequently served as President of his alma mater, Bowdoin College, for 12 years. Despite the fact that he had chronic lower abdominal pain, wound drainage, and a persistent urethral fistula, he lived a very productive life for 50 years after he was wounded at Petersburg. Included in the current paper are commentaries on the state of the art of surgery during the Civil War. PMID- 10818972 TI - An abbreviated history of orthopaedic oncology in North America. AB - Orthopaedic oncology in North America has its roots in European medicine of the 1800s where sarcomas were first classified on the basis of their gross characteristics (1804) and amended on the basis of their histologic features (1867). Surgical treatment, local excision, with unacceptable mortality led to amputation in the 1870s and remained so until limb-sparing resection was cautiously embarked on in the mid-1900s. Nonsurgical adjuvant therapy was first devised in the 1880s (as Coley's toxins) but remained largely ineffective until the advent of chemotherapy in the 1970s. The combination of these techniques in the past 30 years, and the vastly improved staging and reconstructive techniques has led to the current preponderance of limb-salvaging surgery and greatly improved survival rates. The application of these treatments has been enhanced by the development of orthopaedic oncology fellowships, orthopaedic oncology societies, and federally funded regional cancer centers and multidisciplinary teams to treat patients with sarcomas. PMID- 10818973 TI - Rancho Los Amigos Medical Center. A unique orthopaedic resource and teaching institution. AB - Rancho Los Amigos Medical Center, initially a poor farm in the County of Los Angeles, CA became a world renown medical institution because of the polio epidemics in the 1950s. Responding to the need for day to day inpatient care were an overflow of victims of polio who had spine and extremity weakness and were dependent on respirators. Team care, developed at the institution, was used by Vernon L. Nickel, chief orthopaedic surgeon so that maximum use of the limited staff's efforts would be to take care of patients. This need spawned many innovative developments through clinical observations and trials, basic research, and engineering innovations that resulted in the patient's functional improvement and helped return many victims of polio to independence and to their communities. Subsequently, orthopaedic surgeons, Jacquelin Perry, and Alice Garrett joined the full-time staff as the workload increased. Stabilizing the spine using fascial supports, spinal fusion, spinal instrumentation, orthoses, and seating systems allowed those patients who were not totally dependent on respirators to be upright and mobilized. When polio was eradicated, newer programs were established for physically disabled persons with musculoskeletal disorders affecting the spine and extremities and for those patients with congenital, acquired, neurologic, and neuromuscular disorders. In formal graduate residency affiliations, fellowships, and continuing medical education programs orthopaedic surgeons from around the world have been taught the basic principles of "categorical care" for physically disabled people for 50 years. Orthopaedic care given through these programs formed the basis of a new orthopaedic subspecialty, Orthopaedic Rehabilitation. PMID- 10818974 TI - A brief history of the Journal of Bone and Joint Surgery. AB - The Journal of Bone and Joint Surgery had its origins in the Transactions of the American Orthopedic Association, which first were published in 1889. In 1903, the Transactions of the American Orthopedic Association, Volume 16, became the first volume of the American Journal of Orthopedic Surgery, still under the sponsorship of the American Orthopedic Association. In 1919, the word American was dropped from the title and the Journal of Orthopaedic Surgery became the official publication of the newly formed British Orthopaedic Association and the American Orthopedic Association. The name was changed to The Journal of Bone and Joint Surgery in 1921. Shortly after the founding of the American Academy of Orthopaedic Surgeons in 1933, The Journal became the official organ of the academy; however, ownership of The Journal remained with the American Orthopaedic Association. The British volume appeared in 1948. The Journal of Bone and Joint Surgery became an independent, not-for-profit corporation in 1954. The Journal, which continues to be a leader in the area of orthopaedic publication, began publishing the full text on compact disk in January 1992, the full text in electronic form through BRS/Saunders in 1995, and the full text on The Journal's website in December 1999. PMID- 10818975 TI - A century of orthopaedic surgery at Washington University. 1899 to 1999. AB - The 100-year history of orthopaedic surgery at Washington University from 1899 to 1999 is presented. The history focuses on the chairmen, many of whom attained prominence in national orthopaedic organizations, other faculty members, the residency training program, and the affiliated hospitals. Orthopaedic surgery initially was organized as a department, became a division of the Department of Surgery in 1918, and recently again achieved departmental status. PMID- 10818976 TI - The history of the Campbell Clinic. AB - The history of the Campbell Clinic is deeply seated in the accomplishments of Dr. Willis Cahoon Campbell. At the age of 26 he decided to live in Memphis and become an orthopaedic surgeon. At the time Memphis was growing rapidly and orthopaedic surgery was evolving as a surgical specialty. These conditions provided him with the opportunity to do many things that were visionary. He established the Campbell Clinic and the orthopaedic department of the medical school and aided in founding hospitals and clinics for private patients and patients who were indigent. His major contributions to the field of orthopaedics include the recruitment of outstanding associates who would follow and build on his high standards of performance, the establishment of an outstanding residency program, the publication of Campbell's Operative Orthopaedics, and providing leadership for many orthopaedic surgeons. It can be said without reservation that Campbell and the early staff members played an important role in the growth of orthopaedic surgery essentially from its infancy and into the second half of the century. The legacy of excellence that Campbell and all the members of the staff established has continued to be the hallmark of the Campbell Clinic. PMID- 10818977 TI - Arthur Steindler. Orthopaedic scholar, teacher, and clinician. AB - Arthur Steindler, a native of Austria, immigrated to the United States in 1907 and during the next 50 years he became very influential in the development of orthopaedics during the twentieth century. His life is a compelling example of how an individual can make major contributions to the advancement of medical science and practice. His clinical interests spanned the entire range of musculoskeletal disorders including developmental, genetic, infectious, neoplastic and neuromuscular diseases, and their operative and nonoperative treatment. He founded the clinical and academic Orthopaedic Surgery programs at the University of Iowa in Iowa City and directed their growth for 34 years. During that time his clinical work, scholarship, and teaching helped the University of Iowa become a national and international orthopaedic center. He helped establish a statewide system to provide medical care for indigent patients and personally treated more than 70,000 patients, many of them children crippled by polio, tuberculosis, spastic paralysis, scoliosis, congenital deformities, and degenerative neuromuscular diseases. To improve the function of the weakened and deformed limbs of the children, he devised various innovative operations. He wrote more than 130 papers and nine books in several languages on the natural history, etiology, and treatment of musculoskeletal diseases and injuries, pioneered the application of biomechanics to evaluation and treatment of musculoskeletal disorders, and wrote the first comprehensive text on the mechanics of human movement. He taught more than 300 orthopaedists, and when Ruth Jackson, the individual recognized as the first woman orthopaedic surgeon in the United States, decided to pursue a career in orthopaedics, she sought out Arthur Steindler and received her formal orthopaedic education under his direction. That Steindler accomplished so much after moving to the small midwestern town of Iowa City at the age of 37 is impressive, and clearly documents his remarkable abilities and determination. PMID- 10818978 TI - The introduction of arthroscopy to North America. AB - Arthroscopy developed as an offshoot of cystoscopy. Various milestones in the development of endoscopy, and some of the individuals who made significant contributions to the development of arthroscopy are described. After World War II, technology improved and arthroscopy became a valuable adjunct to the armamentarium of the orthopaedic surgeon. There has been a tremendous surge of interest and development of arthroscopy since its reintroduction to North America in 1965. Arthroscopy ranks as one of the three most important contributions to orthopaedics in this past century. PMID- 10818979 TI - The evolution of Shriners Hospitals for Children in North America. AB - Shriners Hospitals for Children have been providing totally free care to children with neuromusculoskeletal disabilities since 1922. This mission expanded in 1962 to include the care of children with burn injuries and expanded in 1978 to include children with spinal cord injuries. Today, Shriners Hospitals for Children include 19 hospitals that provide pediatric orthopaedic care, four hospitals that provide acute and reconstructive burn care, and three hospitals in which spinal cord injury rehabilitation centers are located. During the past 20 years, case acuity and complexity have increased, comprehensiveness of care has been emphasized, members of the medical staff have increasingly become full-time, and sponsored intramural research has increased to a budget of $22 million annually. Annually, more than 250 orthopaedic residents receive pediatric orthopaedic training and more than 150 surgical residents are trained in pediatric burn care in Shriners Hospitals. In 1998, approximately 25,000 children were admitted for care, more than 22,000 operations were performed, and 236,000 children were treated as outpatients. In 1999, the annual operating budget for Shriners Hospitals for Children was $397 million. PMID- 10818980 TI - Partial rupture of the distal biceps tendon. AB - Partial rupture of the distal biceps tendon is a relatively rare event, and various degrees of partial tendon tears have been reported. In the current study four patients with partial atraumatic distal biceps tendon tears (mean age, 59 years; range, 40-82 years) are reported. In all four patients, a common clinical pattern emerged. Pain at the insertion of the distal biceps tendon in the radius unrelated to any traumatic event was the main symptom. In all patients the diagnosis was based on magnetic resonance imaging or computed tomography imaging. In three of four patients the partial rupture of the tendon caused a significant bursalike lesion. The typical appearance was a partially ruptured biceps tendon, with contrast enhancement signaling the degree of degeneration, tenosynovitis, and soft tissue swelling extending along the tendon semicircular to the proximal radius. In three patients, conservative treatment was successful. Only one patient needed surgery, with reinsertion of the tendon resulting in total functional recovery. PMID- 10818981 TI - Bipolar hemiarthroplasty for osteonecrosis of the femoral head. A 7- to 18-year followup. AB - This study evaluated clinical and radiographic results of bipolar hemiarthroplasties for the treatment of osteonecrosis of the femoral head. Forty eight hips in 35 patients with a mean age of 37 years who underwent primary bipolar hemiarthroplasties were observed for an average of 11.4 years. Osteonecrosis was associated with corticosteroid use (21 patients), alcohol (six patients), idiopathic (four patients), and other conditions (four patients). The average Harris hip score was 46 before surgery and 86 at the time of the final followup. Twenty (42%) hips were radiographic failures, and 12 (25%) hips were revised. Groin symptoms were present in 20 (42%) hips. Radiographic proximal migration greater than 4 mm and osteoarthritic signs of the acetabulum indicated a high risk of groin symptoms. The results were inferior to those previously reported for total hip arthroplasty. Thus, for the treatment of osteonecrosis of the femoral head in which necrotic lesions are wide, the authors no longer use this system and currently use total hip arthroplasty. PMID- 10818982 TI - Two- to 9-year outcome after autologous chondrocyte transplantation of the knee. AB - Autologous cultured chondrocyte transplantation was introduced in Sweden in 1987 for the treatment of large (1.5-12.0 cm2) full thickness chondral defects of the knee. The clinical, arthroscopic, and histologic results from the first 101 patients treated using this technique are reported in this study. Patients were assessed retrospectively using three types of endpoints: patient and physician derived clinical rating scales (five validated and two new); arthroscopic assessment of cartilage fill, integration, and surface hardness; and standard histochemical techniques. Ninety-four patients with 2- to 9-years followup were evaluable. Good to excellent clinical results were seen in individual groups as follows: isolated femoral condyle (92%), multiple lesions (67%), osteochondritis dissecans (89%), patella (65%), and femoral condyle with anterior cruciate ligament repair (75%). Arthroscopic findings in 53 evaluated patients showed good repair tissue fill, good adherence to underlying bone, seamless integration with adjacent cartilage, and hardness close to that of the adjacent tissue. Hypertrophic response of the periosteum or graft or both was identified in 26 arthroscopies; seven were symptomatic and resolved after arthroscopic trimming. Graft failure occurred in seven (four of the first 23 and three of the next 78) patients. Histologic analysis of 37 biopsy specimens showed a correlation between hyalinelike tissue (hyaline matrix staining positive for Type II collagen and lacking a fibrous component) and good to excellent clinical results. The good clinical outcomes of autologous chondrocyte transplantation in this study are encouraging, and clinical trials are being done to assess the outcomes versus traditional fibrocartilage repair techniques. PMID- 10818983 TI - Magnetic resonance arthrography in children with developmental hip dysplasia. AB - Conventional and gadolinium enhanced magnetic resonance arthrograms were done on 14 hips in 10 children ages 7 to 24 months. The contralateral normal hips in those with unilateral disease were studied with unenhanced magnetic resonance imaging for comparison. By conventional arthrography, there were no well visualized structures. Visualized structures seen as filling defects were the labrum, ligamentum teres, and transverse acetabular ligament. By magnetic resonance arthrography, well visualized structures were the labrum, ligamentum teres, transverse acetabular ligament, and pulvinar. By unenhanced magnetic resonance imaging, well visualized structures were the labrum, ligamentum teres, and transverse acetabular ligament. The difference in visualization by magnetic resonance arthrography versus conventional arthrography was statistically significant with respect to all five structures: labrum, ligamentum teres, transverse acetabular ligament, pulvinar, and psoas tendon. The difference in visualization by magnetic resonance arthrography versus unenhanced magnetic resonance imaging was statistically significant with respect to the labrum and pulvinar. Magnetic resonance arthrography is indicated for assessing complete concentric reduction when it does not appear to be achieved by conventional arthrography, for confirming closed reduction immediately after manipulation, and potentially for preoperative planning for an open reduction. PMID- 10818984 TI - Nonrhabdomyosarcoma soft tissue sarcomas in children. The Mayo Clinic experience. AB - Eighty-six children to 18 years of age were treated for nonrhabdomyosarcoma soft tissue sarcomas of the trunk and extremities. Synovial sarcoma (31), fibrosarcoma (13), malignant fibrous histiocytoma (11), epithelioid sarcoma (10), and clear cell sarcoma (7) were the most common diagnoses. Four patients presented with metastatic disease. A high percentage of patients presented after biopsy by the referring physician, although this could not be shown to affect outcome. Patients were treated with wide removal of the tumor when possible, with judicious use of adjuvant radiation, or with chemotherapy in selected cases. Mean followup was 11 years. Five- and 10-year survival was 92% and 84%, respectively. Tumors larger than 5 cm were associated with a worse prognosis. When compared with published data in adults, the prognosis of primary, localized nonrhabdomyosarcoma soft tissue sarcomas in children appears to be more favorable. PMID- 10818985 TI - Impact of postoperative gait analysis on orthopaedic care. AB - The impact of postoperative gait analysis on the ongoing orthopaedic care of 38 consecutive patients with a static encephalopathy was evaluated. Of the 38 postoperative gait analyses, 32 (84%) resulted in recommendations of a change in patient care. Surgery was recommended in 16 of 38 (42%) cases, bracing in 20 (53%) cases, and specific physical therapy regimens in eight (21%) cases. Eleven of the 38 (29%) patients had changes recommended in at least two of the three areas (surgery, bracing, and therapy). The results of this study suggest that postoperative gait analysis serves not only as a measure of treatment outcome, but also as a useful tool in planning ongoing care for these patients. PMID- 10818987 TI - Anatomy of the iliocapsularis muscle. Relevance to surgery of the hip. AB - Gross anatomic dissections of 20 fresh adult human anatomic hip specimens were performed to characterize the anatomy of the iliocapsularis muscle, a little known but constant muscle overlying the anteromedial hip capsule. This muscle originates in part from the inferior border of the anterior-inferior iliac spine, but the main origin arises from an elongated attachment to the anteromedial hip capsule and inserts just distal to the lesser trochanter. This muscle is an important landmark for exposure of the anteromedial hip capsule and psoas tendon interval during performance of the Bernese pericapsular osteotomy. It is postulated that contraction of this muscle theoretically can tighten the anterior hip capsule, thus helping to stabilize the femoral head within a dysplastic acetabulum. PMID- 10818986 TI - Spatial orientation of the microscopic elements of cortical repair bone. AB - This study was designed to assess whether the original orientation of the elements of cortical diaphyseal bone is reestablished during 18 months of repair. Full thickness 3-mm drill holes were created surgically through the tibial and femoral cortices of adult rabbits and studied by light and polarizing microscopic analysis at 1, 4, 8, and 16 weeks and 6, 12, and 18 months after the defects were created. From 16 weeks to 12 months the defects were repaired fully with lamellar bone, while the longitudinal spatial orientation of the repair lamellae was at a distinct angle to that of the original bone. At 18 months, the internal organization of the major individual microscopic elements (osteons) were reconstituted in a manner similar to that of the original bone; however, the three-dimensional spatial orientation of the osteons in the repair bone relative to the long axis of the osteons of the adjacent original bone and of the bone removed to create the defect was not reestablished completely. The imperfect spatial disposition of the microscopic elements of bone tissue may contribute significantly to refracture of a long bone after removal of screws from plated or unplated long bone fractures. PMID- 10818988 TI - Effects of monopolar radiofrequency energy on ovine joint capsular mechanical properties. AB - Radiofrequency energy may provide a relatively noninvasive method to stabilize joints with excessive laxity by thermally shrinking redundant joint capsular tissue. The authors determined the percentage of shrinkage associated with five radiofrequency treatment temperatures and evaluated the effect of this energy on the structural properties of joint capsular tissue in vitro. First, 36 adult sheep femoropatellar joint capsular specimens were treated with one of five treatment temperatures (n = 6 per group) or served as a control to determine tissue shrinkage. An additional 24 specimens were treated with three temperatures that resulted in different shrinkage: 45 degrees C, 65 degrees C, and 85 degrees C. Tissue stiffness, relaxation, and failure strength were determined for each specimen (n = 6 per group). Tissue shrinkage was correlated significantly with treatment temperature. There was a significant decrease in tensile stiffness in the 65 degrees C and 85 degrees C treatment groups. There were no significant differences between stress relaxation before treatment and after treatment. Relaxation properties after treatment were not different from each other or from control values either normalized to pretreatment values or expressed as raw data. Failure strength was not affected significantly at any temperature. PMID- 10818989 TI - Multiple lytic skeletal lesions and hypercalcemia in a 13-year-old girl. PMID- 10818990 TI - Osteogenic induction in hereditary disorders of heterotopic ossification. AB - The formation of heterotopic bone within soft connective tissue is a common feature of at least three distinct genetic disorders of osteogenesis in humans: fibrodysplasia ossificans progressiva; progressive osseous heteroplasia; and Albright hereditary osteodystrophy. The pathobiologic characteristics of osteogenic induction, the histopathologic features of osteogenesis, the anatomic distribution of heterotopic lesions, and the developmental patterns of disease progression differ among all three conditions. The molecular and cellular basis of redirecting a mature connective tissue phenotype to form bone is a remarkable biological phenomenon with enormous implications for the control of bone regeneration, fracture healing, and disorders of osteogenesis. PMID- 10818991 TI - [Is there an age limit for heart catheterization?]. PMID- 10818992 TI - [Sublingual captopril: indications and advantages?]. PMID- 10818993 TI - [Negative inotropic effects and constipation caused by verapamil?]. PMID- 10818994 TI - [Role of nitrates in the treatment of ischemia]. PMID- 10818995 TI - [Development of nitrate tolerance]. PMID- 10818996 TI - [Paroxysmal atrial fibrillation]. PMID- 10818997 TI - [What anticoagulant therapy should be used for what form of atrial fibrillation?]. PMID- 10818998 TI - [Anticoagulation after successful cardioversion in atrial fibrillation?]. PMID- 10818999 TI - [Prevention of embolism in atrial septal defects?]. PMID- 10819000 TI - [Hypertensive microangiopathy]. PMID- 10819001 TI - [Endomyocardial biopsy in myocarditis]. PMID- 10819002 TI - [Ebstein's anomaly]. PMID- 10819003 TI - [Pharmacological bases of circulation-stimulating effect of champagne]. PMID- 10819004 TI - [When must an indwelling venous catheter be flushed?]. PMID- 10819005 TI - [Theme emphasis for this issue--cardiology]. PMID- 10819006 TI - [Treatment for arrhythmia today--is there an optimal method?]. PMID- 10819007 TI - [Focal paroxysmal atrial fibrillation. Experiences with treatment using high frequency catheter ablation]. AB - BACKGROUND AND OBJECTIVE: As has recently been discovered, paroxysmal atrial fibrillation (PAF) can be induced by focal extrasystoles or tachycardia. This raises the question of whether this form of atrial fibrillation can be cured by high-frequency catheter ablation of the focal trigger. PATIENTS AND METHODS: Seven men and eleven women (mean age 45.6 +/- 11 years) with severe symptoms and treatment-resistant PAF underwent electrophysiological tests with the aim of high frequency catheter ablation, if long-term ECG monitoring had demonstrated frequent atrial extrasystoles or tachycardia as pointer to a focal origin. Ablation was performed at the point of earliest excitation after the origin of the ectopic focus had been localized. The end-point was reached if the atrial ectopic rhythm had ceased for more than 60 min. RESULTS: In 18 of the 20 patients an ectopic focus was found and successfully ablated (1 in the superior vena cava, 3 in the right atrium and 16 in a pulmonary vein). Atrial ectopic beats recurred within 24 hours of ablation in 6 of the 14 patients with a pulmonary vein focus: a second focus was found in two, re-emergence of the original focus in two, no re investigation in another two. 13 of the 18 patients have had no further symptoms after a mean follow-up of 11 months without anti-arrhythmia treatment. CONCLUSION: The results indicate that focally induced paroxysmal atrial fibrillation can be cured by locally applied high-frequency ablation. PMID- 10819008 TI - [Is the estimation of the progression of valvular aortic stenosis possible?]. AB - BACKGROUND AND OBJECTIVE: It is of great importance to assess progression of aortic valvar stenosis (AVS) when cardiac surgery is planned for other indications when established criteria for aortic valve replacement are not fulfilled at that moment. These considerations have often been ignored in prospective planning of treatment, necessitating a second cardiac surgical intervention just a few years later. The aim of this study was to establish criteria for estimating the rate of progression of AVS. PATIENTS AND METHODS: Clinical, echocardiographic and haemodynamic data were analysed for 169 patients with aortic valvar stenosis (169 men, 88 women; mean age at first cardiac catheterization [CC] 55.2 +/- 15.7 years, at second CC 63.4 +/- 15.6 years. RESULTS: The degree of AVS increases exponentially in relation to the extent of calcification (graded 0-3) and the fall in transaortic gradient (TG), from a TG > 0.6 mmHg/ml stroke volume and can be sufficiently predictable for clinical purposes. But neither age, sex nor the aetiology/pathology of the valvar defect have a sustained influence on the progression of AVS. CONCLUSIONS: These data indicate that knowing the current reduction in TG and the degree of calcification makes it possible to assess the likely progression of previously asymptomatic AVS and thus greatly facilitate the decision of whether or not to combine aortic valve replacement with another indicated cardiac operation. PMID- 10819009 TI - [Diagnosis and therapy of an intramural hematoma of the ascending aorta]. AB - HISTORY AND CLINICAL FINDINGS: A 79 year-old female presented with severe chest pain and shortness of breath. INVESTIGATIONS: The chest X-ray showed a widened mediastinum and a small left-sided pleural effusion. Transesophageal echocardiography demonstrated not only dilatation of the ascending aorta and aortic regurgitation but also an echo-poor crescentic seam within the aortic wall. Computed tomography identified this structure as an intramural hematoma. TREATMENT AND COURSE: The aortic valve and the ascending aorta were replaced. CONCLUSION: An intramural hematoma of the ascending aorta may be difficult to recognize but it is a life-threatening condition and should be treated by prompt surgical repair. PMID- 10819010 TI - [Myocardial revascularization for therapy-refractory angina pectoris in the elderly]. PMID- 10819011 TI - [Vasopeptidase inhibitors. Clinical implications of a new class of drugs]. PMID- 10819012 TI - [Drug therapy of cardiac arrhythmias]. PMID- 10819013 TI - [Interventional therapy of cardiac arrhythmias]. PMID- 10819014 TI - Genetic and biochemical studies with ataxia telangiectasia. A review. AB - This article summarizes the genetics and clinical features of ataxia telangiectasia (AT) and then reviews recent cytogenetic, cellular, and biochemical studies which support the hypothesis that a defect in DNA repair is responsible for the various manifestations of the disease. The biochemical evidence further indicates that the defect specifically reduces the cellular capacity to remove bases and nucleotides damaged by ionizing radiation, without affecting the cells' ability to scavenge free radicals or to rejoin breaks in the sugar-phosphate backbone of DNA. Suggestions for additional research to more precisely identify the repair defect will also be presented. PMID- 10819015 TI - Preferential fluorescent staining of heterochromatic regions in human chromosomes 9, 15, and the Y by D 287/170. AB - The utility of a newly synthesized chemical variation of DAPI (4'-6-diamidino-2 phenyl-indole), D 287/170, for differential staining of constitutive heterochromatin in man is demonstrated. Direct staining of human chromosomes with D 287/170 results in brilliant fluorescence of the paracentromeric C-band of chromosome 9, of a proximal short-arm segment of chromosome 15, and of certain heterochromatic regions in the Y. Bright, but less conspicuous fluorescence is occasionally seen at the centromeres of other chromosomes. The staining differentiation obtained by D 287/170 is very distinct, and the intensity of the fluorescent light is unusually high. The new fluorochrome should prove particularly useful for detecting and analyzing human chromosome 9 heterochromatin at various stages of the cell cycle in normal and structurally altered chromosomes. PMID- 10819016 TI - Variation in pattern and frequency of acrocentric association in normal and trisomy-21 individuals. AB - Chromosomally normal and trisomy-21 individuals were studied for the ability of their nucleolus-organising chromosomes to form satellite associations in G-banded lymphocyte metaphases. Two types of parameter, absolute association frequency and relative association frequency, were used. There was no significant difference between females and males or between Caucasoids and Mongoloids for either type of association parameter in the controls, nor was there significant correlation between age (17-40 years) and either type of parameter in the controls. The pattern of two chromosome associations is accounted for by two related models in both normal and trisomic individuals. These models imply that there is an extensive polymorphism for associating ability and that this ability may be zero in individual chromosomes. Homologous do not associate preferentially with each other. The absolute frequency of acrocentric association is lower in trisomy 21 individuals than disomic controls, but the relative involvement of chromosome 21 (after correction for the trisomic state) is higher than in the controls. PMID- 10819017 TI - X-linked mental retardation with fragile X. A pedigree showing transmission by apparently unaffected males and partial expression in female carriers. AB - A large family is reported in which mental retardation associated with the fragile site at Xq28 was found. Three normal males seemed to have transmitted the trait through their daughters to affected grandchildren. A total of 19 family members were investigated cytogenetically. Mentally retarded males showed macroorchidism and the fragile X. Three mentally retarded females were found, with the fragile X in a high percentage of cells; in contrast, the obligate carriers showed no or only few cells with the fragile X. PMID- 10819018 TI - Characterization of a new aberration of the human Y chromosome by banding methods and DNA restriction endonuclease analysis. AB - Comparative cytogenetic analyses were performed with ten different banding methods on a previously undescribed, inherited structural aberration of a Y chromosome, and the results compared with those of normal Y chromosomes occurring in the same family. The value of the individual staining techniques in investigations of Y chromosomal aberrations is emphasized. The aberrant Y chromosome analyzed can be formally derived from an isodicentric Y chromosome for the short arm with a very terminal long-arm breakpoint, in which the centromere, an entire short arm, and the proximal region on one long arm was lost. This interpretation was confirmed by determining the amount of the two Y-specific DNA sequences (2.1 and 3.4 kb in length) by means of Hae III restriction endonuclease analysis. The karyotype-phenotype correlations in the men with this aberrant Y chromosome, especially the fertility dysfunctions (oligoasthenoteratozoospermia, cryptozoospermia), are discussed. The possibility of the existence of fertility factors involved in the control of spermatogenesis within the quinacrine-bright heterochromatic region of the Y long arm is presented. PMID- 10819019 TI - Isoelectric focusing of red cell phosphoglucomutase (E.C.: 2.7.5.1) at the PGM1 locus in a French-Canadian population. AB - Phosphoglucomutase1 (PGM1) polymorphism was studied in a French-Canadian population of Quebec city, Canada by means of a low voltage (max 500 V) isoelectric focusing (IEF) procedure on vertical polyacrylamide gel slabs. Frequencies of the four common PGM1 genes estimated from the phenotype distribution in 308 unrelated individuals were PGM1(1+), 0.61 (+/- 0.02); PGM1(1 ), 0.13 (+/- 0.01); PGM1(2+), 0.18 (+/- 0.02); and PGM1(2-), 0.08 (+/- 0.01). The segregation patterns observed in 154 families, which included 31 different mating types and 353 children, confirmed a Mendelian inheritance of four autosomal genes. The distribution of the PGM1 phenotypes observed or expected in a Hardy Weinberg equilibrium was compared with that of other populations. A significant (P < 0.001) difference was found between the Quebec population and a Black population from Keneba, Gambia, West-Africa. PMID- 10819020 TI - Familial factors in early deaths: twins followed 30 years to ages 51-61 in 1978. AB - Subjects in the National Academy of Sciences-National Research Council Twin Registry of 31,848 male twin veterans were followed for mortality from 1 January 1946, or from the date of entry into military service if that was later, to 31 December 1978. During this time 3,573 deaths occurred among them, 837 due to trauma and 2,712 due to disease. Mortality from all causes for the entire follow up period was 10.2% among 11,350 monozygotic (MZ) twins and 11.4% among 14,450 dizygotic (DZ) twins. Mortality of veterans is known to be favorable compared to U.S. males. Among U.S. males of the same ages as the two respective twin zygosity groups, a mortality of 13.9% would have been expected during this time period. Observed mortality from trauma was 2.3% for MZ twins and 2.5% for DZ twins, with 3.0% expected in either group. Observed mortality from all disease was 7.9% for MZ twins and 8.8% for DZ twins, with 10.9% expected in either group. For total mortality, the case twin concordance rates, based on individual deaths, were 28.2% among MZ twins and 17.7% among DZ twins. For trauma, respectively by zygosity, these concordance rates were 6.9% and 3.9%. In this sample, familial factors appear to be of little consequence in trauma deaths. For all disease the concordance rates were 30.1% and 17.4%. Estimating heritability of liability to death from disease, as proposed by Edwards (1969), provides values of h2 = r = 0.51 for MZ twins, h2 = 2r = 0.48 for DZ twins, and h2 = 2(rMZ-rDZ) = 0.54 using data for the two zygosity groups combined. PMID- 10819021 TI - [Electrophoretic polymorphism of post-albumin and transferrin in different subspecies of anthropoid apes]. AB - The electrophoretic polymorphism of post-albumin and transferin was investigated in 46 apes of the Pan troglodytes, Gorilla gorilla and Pongo pygmaeus subspecies. This type of study enables the different subspecies to be distinguished and the ancestral form to be identified. The pattern of individual variability is found to be different between the various ape species and Homo sapiens. PMID- 10819022 TI - Genetic disposition to alcoholism. An EEG study in alcoholics and their relatives. AB - Family, twin, and adoption studies have shown that genetic factors are involved in the etiology of alcoholism. Based on earlier EEG findings in alcoholics and on the known genetic determination of the alcohol effect on the EEG, the hypothesis was tested whether the resting EEG reflects a certain disposition to alcoholism. Resting EEGs were examined for 115 alcoholics (78 males, 37 females) and matched controls. In addition, the first-degree relatives of two extreme groups of alcoholics--those with poor and those with particularly good alpha waves--were examined and compared with matched controls. The EEGs were analyzed with an EEG processor. Whereas male alcoholics did not differ from their controls, female patients showed a shift from the alpha and theta to the beta bands of the brain wave pattern. The relatives of the two extreme groups of alcoholics, who did not misuse alcohol, exhibited the same tendency. This is an argument supporting the notion that in females a poorly synchronized EEG pattern reflects a certain disposition to alcoholism. This finding is discussed in light of drinking motivation in males and females. The latter more often belong to the alpha- and gamma-types of alcoholism than do males. Because of comparable findings in schizophrenics it is argued that a genetically determined desynchronized resting EEG pattern is not specific for a certain illness, but reflects basic mechanisms that enhance the risk for different psychiatric disorders. PMID- 10819023 TI - Human transferrin (Tf) and group-specific component (Gc) subtypes in Tunisia. AB - Simultaneous subtyping of two genetic markers--group-specific component (Gc) and transferrin (Tf)--by electrofocusing enabled us to compute the following gene frequencies for the Tunisian population: Gc1S, 0.525; Gc1F, 0.260; Gc2, 0.215; TfC1, 0.770; TfC2, 0.215; TfD1, 0.015. The frequencies of TfD, TfC2, and Gc1 are higher than those found in Caucasoid populations and can be explained by Negroid contribution. A selective advantage related to the metabolic role of this vitamin D-binding protein does not seem very likely for any particular Gc type or subtype. It is postulated that the differences in the frequencies of the Gc alleles might be related to selective advantage for genes belonging to other genetic systems originally closely linked to either Gc1 or to Gc2 alleles. PMID- 10819024 TI - Complex segregation analysis of the locus for beta-aminoisobutyric acid excretion (BAIB). AB - High excretion of BAIB is determined by an incompletely recessive gene jointly with other familial factors. The parameters estimated from a Japanese sample are: gene frequency = 0.6, displacement between homozygotes = 2.1 sigma, dominance = 0.2, and polygenic heritability = 0.1. Supplementary tests support this mixed model and (unlike classical analysis) give no evidence of segregation disturbance. Complexity of BAIB inheritance should be allowed for in population comparisons and linkage studies, which can be performed reliably under the mixed model. PMID- 10819025 TI - Analysis of the red cell membrane in a family with hereditary elliptocytosis- total or partial of protein 4.1. AB - In a 12-year-old boy carrying a clinically silent elliptocytosis, we observed a total lack of red cell membrane band 4.1. Band 4.1 was partially absent in the father who also displayed a clinically silent elliptocytosis and, remarkably, in the mother although she presented normal discocytes. Band (2 and 2.1.) phosphorylation was sharply reduced in the three persons examined. In the propositus and his mother, but not in his father, a clearly phosphorylated band appeared at the level of band 4.2. We suggest that the father and the mother carry two distinct alleles affecting differently the interactions within the spectrin-actin protein 4.1 complex. The father's allele is elliptocytogenic in the heterozygous state and, among other molecular alterations, prevents the attachment of protein 4.1. The mother's allele is morphologically silent in the heterozygous state, yet it also affects the binding of protein 4.1, possibly because the latter is shortened. The propositus, being doubly heterozygous, has the same morphological phenotype as his father, but his protein 4.1 electrophoretic phenotype is the addition of both parental phenotypes. The distinct phosphorylation patterns in the region of bands 4.1 and 4.2 are also consistent with the two-allele hypothesis. PMID- 10819026 TI - Group-specific component (Gc) subtypes in the Chinese population of Hong Kong. AB - The distribution of Gc subtypes in a sample of the Chinese population of Hong Kong was studied using isoelectric focusing followed by immunofixation. A sensitive modification of this technique is described. Nine distinct phenotypes were observed which appear to result from the three common alleles Gc1F, Gc1S, and Gc2, which are found in most populations. The respective gene frequencies were 0.494, 0.258, and 0.247. In addition, two rare phenotypes were observed which appear to be due to a rare allele tentatively identified as Gc2C2. PMID- 10819027 TI - Gc subtyping in Malaysians and in Indonesians from north Sumatra. AB - Malays, Chinese and Indians from peninsular Malaysia; Ibans and Bidayuh from Sarawak state, Northern Borneo; and Bataks, Minangkabau and Javanese from North Sumatra, Indonesia, were subtyped for Gc (group-specific component) by polyacrylamide gel isoelectric focusing. All eight populations investigated were found to be polymorphic for three common alleles, Gc1F, Gc1S and Gc2. PMID- 10819028 TI - Red cell antigen, serum protein, and red cell enzyme polymorphisms in inhabitants of the Jimi Valley, Western Highlands, New Guinea. AB - A series of blood samples from four villages in the Jimi Valley, Western New Guinea Highlands, has been tested for genetic variation in blood group, serum protein, and red cell enzyme systems. Polymorphic variation was present for the AB0, MNS, P, and Rh blood group systems, for the Hp and Tf serum protein systems, and for the acid phosphatase, 6-PGD, PGM, MDH, and ADA enzyme systems. One each of the following variants was detected: Ge(a-), G6PD deficient, AK 2-1 and PHI 7 1 or 8-1. All samples tested were Cw-, K-, Kp(a-), Wr(a-), Fy(a+ b-), Rd-, and LDH normal. Genetic distance analysis places the Jimi Valley populations closer to peoples of the Chimbu-Chuave and Wahgi-Hagen areas than to the Maring people of the Simbai Valley to the north. PMID- 10819029 TI - Rare phenotypes of the phosphoglucomutase locus 1 detectable by isoelectric focusing on Cellogel. AB - The rare phenotypes PGM1, determined by alleles PGM1(3), PGM1(4), PGM1(6), and PGM1(7) were examined by starch gel electrophoresis and cellulose acetate gel isoelectric focusing and were compared with the commonest phenotypes of PGM1. The frequencies of the rare genes found in the Polish populations were as follows: in Lublin, PGM1(3) = 0.0002, PGM1(4) = 0.0005, PGM1(6) = 0.0010, and PGM1(7) = 0.0005; in Wroclaw, PGM1(3) = 0.0000, PGM1(4) = 0.0005, PGM1(6) = 0.0007, and PGM1(7) = 0.0002. The results suggest that the F and S type variants of the genes PGM1(4) and PGM1(7) probably do not occur. It is still possible that F and S variants exist for the genes PGM1(3) and PGM1(6). PMID- 10819030 TI - Discrepancy between G and R bands. Example of an acute non-lymphocytic leukemia. AB - An apparently different chromosome abnormality was observed in unstimulated blood cultures from an acute non-lymphocytic leukemic child: 11q- with G banding techniques and 17q- with R banding techniques. The abnormality is explained as a t(11;17) translocation, and the discrepancy between the G- and R-band patterns discussed. PMID- 10819031 TI - Steroid sulphatase levels in XX males, including observations on two affected cousins. AB - Quantitative assays of steroid sulphatase in XX males have shown that some individuals have two functional loci, and others only one. Two affected cousins, who cannot share the same X-chromosome, nevertheless have male levels of steroid sulphatase, suggesting functional abnormality of the X chromosome. The hypothesis is advanced that these and other unusual features of X-chromosome function in some XX males, could be explained if such cases were due to an autosomal mutation, exercising its effect by causing abnormal inactivation of a subterminal area of Xp which normally escapes the inactivation process. PMID- 10819032 TI - Frequency of human A gamma 75Thr globin chain in a population from Tunisia. AB - Cord blood samples, collected at Sousse and Monastir, from Tunisian newborns were focused on a thin layer of agarose in order to detect the carriers of the A gamma 75Thr chain (A gamma chain bearing a replacement Ile-->Thr at position 75). Nineteen individuals (10%) were positive for this variant. The frequency of the A gamma 75Thr gene in the Tunisian population (0.050) is compared with that of various ethnic populations. PMID- 10819033 TI - [The role of adsorbed water in polysaccharide radiation destruction]. AB - The role of water in the potato starch destruction during gamma-irradiation of native starch preparations with various equilibrium humidity (5-45%) and starch jellies (systems with different quantity of polymer structured water) has been studied. The scheme reactions of the formation of primary radicals and molecular products of radiolysis in the system "structured starch--water" is also given. PMID- 10819034 TI - [Structure-functional states of liver mitochondrial membranes of rats exposed to irradiation]. AB - The effect of the low dose gamma-irradiation (270 cGy--one-fold; 90 cGy per day during 3 days) on oxidative phosphorylation, lipid peroxidation, microviscosity of the annular and free lipids membrane, and membrane protein structural state was studied. The post-radiation influence on membrane functional activity and structural state in accordance with the irradiation regimes was established. PMID- 10819035 TI - [Effect of age and radiation exposure on the frequency of translocations and dicentrics detected by FISH method in human lymphocytes]. AB - The frequencies of translocations and dicentrics detected by "chromosome painting" in lymphocytes were estimated in 115 healthy donors and in 273 people exposed to uncontrolled irradiation at low doses 1-4 years ago. Age responses of both types of exchanges at the age range from 3 to 85 years fit to quadratic model. The frequency of translocations grew faster with age than the frequency of dicentrics. The yields of stable exchanges in exposed people was significantly higher than those in control donors of corresponding ages. PMID- 10819036 TI - [On the mechanism of cytogenetic effect of electromagnetic radiation: a role of oxidation homeostasis]. AB - It was established in the experiments on rats that the changes in free radical oxidation under the influence of non-ionizing radiation had a wavy character. It was revealed that the changes in oxidation homeostasis preceded development of cytogenetic effects and could be their reason. PMID- 10819037 TI - [The effect of small doses of ionizing radiation on the physico-chemical characteristics of the membranes of peripheral blood lymphocytes of rats]. AB - The fluorescence probe method was used for investigating the physical state of a total lipid phase of a bi-layer and an annular (near-protein) zone of the membrane lipids of lymphocytes in a peripheral blood of rats on the 10th day after a whole-body acute and chronic gamma-exposure to doses of 0.25, 0.5, and 1.0 Gy. It was discovered that exposure to doses of 0.25, 0.5, and 1.0 Gy revealed no reliable distinctions in the parameters of a physical state of the lipid component of membranes within the given period of observation if compared with those of controls. However chronic exposure to the same doses caused the increase in hydrophobicity of the total lipid phase of the membrane bi-layer with no change in the polarity of an annular lipid. The near-protein zone of lipids revealed a local decrease in microviscosity while the fluidity of total lipids of a membrane bi-layer remained unaltered. A detected change of tryptophan fluorescence of the membrane proteins after exposing them to small dosed has also been carried out. PMID- 10819038 TI - [Effect of small doses of ionizing radiation on the level of catecholamines and corticosteroids in mouse adrenals]. AB - Effects of 137Cs gamma-radiation (0.06-0.54 cGy, 0.06 cGy/day) on the levels of catecholamines and corticosteroids in mouse adrenals were investigated. There were observed increase of these parameters after mice irradiation during 1-2 days and their decrease after mice irradiation during 9 days. PMID- 10819039 TI - [The role of antioxidative status in development of the consequences of biological action under low dose and low intensity irradiation]. AB - The scale and direction of changes of a number of biophysical, biochemical and physiological parameters in SHK mice (male) tissues were studied depending on their initial values for mice control groups within 1 day after low intensity gamma-irradiation (15 cGy). The reciprocal dependences between the scales of the lipid antioxidative activity (AOA) in brain or the spleen mass changes and their values for mice control groups were found. The external dependences were revealed between the amounts of lipid peroxidation secondary products in spleen, liver and blood plasma of the control mice and a scale of their change after irradiation. The most substantial changes of this parameter were observed in blood plasma and the changes of the phospholipid content within the total lipid composition were found in spleen and blood erythrocytes of the irradiated mice, that is in the lipids of tissues, which had the lowest level of AOA for mice control groups. The experimental data obtained indicate that the initial antioxidant status of animal tissues plays the important role for the development of consequences of the biological action under low dose and low intensity irradiation. PMID- 10819040 TI - [Effect of low doses of ionizing radiation on thymus-dependent humoral immune response and the polyclonal activation of B lymphocytes]. AB - The exposure of mice to low dose of gamma-rays (10 cGy, 1 cGy/min) increased thymus-dependent humoral immune response and polyclonal activation of B-cells. Injection of hydroquinone eliminated radiation-induced augmentation of polyclonal response of B-lymphocyte. Thymogen decreased significantly the radiation-induced immunostimulation. The study of the dynamics of primary immune response showed that the period of radiation-induced elevation was followed by the phase of profound reduction of antibody formation. Possible negative consequences of action of low doses of ionizing radiation on immune system is discussed. PMID- 10819041 TI - [Comparative od adaptive response to gamma-radiation and nickel sulfate treatment in human cells]. AB - Increased viability of human cells (line rhabdomyosarcoma) to challenge doses of NiSO4 (10(-5)-10(-3) M) was formed when cells were preirradiated with low doses of gamma-radiation (10-14 cGy). Observed adaptive response was similar to radioadaptive response in human fibroblasts, pretreated with low doses of gamma radiation and challenge dose of the same mutagen. Pretreatment with low concentration of NiSO4 induced in human fibroblasts increased resistance of DNA to the treatment with challenge doses of gamma-radiation and stimulated DNA repair synthesis after treatment with NiSO4 and 4-nitroquinoline-1-oxide. These data confirm the existence of cross-adaptation in the experiments with NiSO4. PMID- 10819042 TI - [Clinico-immunological and allergological characteristics of population of the Middle ural]. AB - Clinical investigation of immune and allergic state of 633 adult dwellers of the Middle Ural region was carried out. The first (occupational) group consisted of 479 workers of the industrial plant, most of which exposed to negative factors such as radiation, harmful chemical substances and stress. The second (control) group consisted of 154 town-dwellers, who did not work at the industrial plant. Immune state was assessed in 714 people. Clinical symptoms of immune deficiency were found in 67% of the occupational group and in 76% of the control group. Allergic condition and allergic-infection syndrome were common for both groups, the rate of infection syndrome being relatively low. The immune state of the occupational group showed in comparison to control reliable increase in absolute and relative amount of T-lymphocytes, in the ratio CD4+/CD8+ and in the level of serum IgG. It was found also the decrease in the concentration of T-killers, the amount of NK cells and B-lymphocytes, phagocyte activity and the level of serum IgA. It was suggested that regional and ecological peculiarities influence the immune state rather than induce clinical symptoms of immune deficiency. PMID- 10819044 TI - [Possibility of prediction of synergistic effects at a whole organism level]. AB - The possibility of applying the mathematical model of synergism, developed earlier for description of combined effects in cell system, to the description of synergism on organisms level was developed. The model describes data on combined action of ionising radiation and other harmful environmental agents on survival and cancerogenesis in laboratory animals and human beings. PMID- 10819043 TI - Comparative analysis of proinflammatory cytokines in plasma of mice exposed to radiation or in combined radiation injury. AB - Male mice (CBA x C57BL/6)F1 were used for the experiments throughout this study. Blood serum levels of IL-1 beta, IL-6, TNF-alpha, IL-3, and GM-CSF were evaluated by means of enzyme-linked immunosorbent assay at 3, 6, 24, 48 and 72 hours after 7 Gy gamma-irradiation alone or combined injury (irradiation + thermal burn). Radiation as well as combined injury did not cause any important alterations of IL-1 beta, TNF-alpha, IL-3, and GM-CSF concentrations in the system circulation. Combined injury revealed more enhanced serum levels of IL-6 versus only irradiated mice. A possible significance of this phenomenon at the combined radiation and thermal burns' pathogenesis is discussed. PMID- 10819045 TI - [Immunological mechanisms of the radioprotective effects of ceruloplasmin]. AB - The aim of the work was to examine immunotropic features of celluroplasmine++ (Cp) and estimate its capacity as radioprotector. The pulse radiolysis method was used to set the mechanism of elementary reactions, which were responsible for antioxidant activity, and to demonstrate a particular role of reversible oxycomplexis, Cp...O2. Within in vitro test-systems the effects of Cp were examined on the model of interaction between lymphocytes from Shelter staff which constantly contacts with ionizing irradiation, and autologous erythrocytes in rosset-forming phenomenon (223 Shelter employees in comparison with 253 donors of Kyiv blood transfucion station). Expression of this Index in the presence of antigens with known poly-specificity was determined (antigens were taken from cortical and pyramid sections of kidney, liver, lungs, myocard, pancreatic gland, grey matter and aorta). Simultaneously the presence of analogous autoantibodies was determined (ELISA). It has been shown that Cp can reprogram the level of expression of immunovaluable receptors towards to norm. It has been also defined that Cp presence in blood irradiated in vitro (1-15 rem) promotes the masking of active centers of autoantibodies of different tissue specificity. PMID- 10819046 TI - [Radiosensitization of human tumor HeLa cells induced by methyl ester of 6-oxo-6 [2,2-ethylenedioxy-5-(dimethoxycarbonylmethyl)-cyclope nt- 1-yL]-hexanoic acid]. AB - Methyl ester of 6-oxo-6-[2,2-ethylenedioxy-5-(dimethoxycarbonylmethyl)- cyclopent 1-yl]-hexanoic acid (7-keto-9,9-ethylenedioxiprostanoid, that is analogue of 11 deoxy-PGE1 with modified chains) at concentration 10(-6) mol/l displayed maximal (25%) sensitizing effect when was tested in a range of concentrations 10(-7)-10( 5) mol/l and HeLa cells were gamma-irradiated at dose of 2 Gy, that was compared with metronidazol action. Optimal time of prostanoid contact with cells was between 30 and 60 min, and metronidazol--60 min. Prostanoid effect was exhibited at irradiation doses from 2 to 4 Gy and reached maximum of 46% at 4 Gy, its DMF was 1.5 as well as that of metronidazol. Cytotoxicity of comparable compounds was approximately equal. By its chemical structure and sensitizing features this 7 keto-9,9-ethylenedioxiprostanoid is a prototype of new radiosensitizing drug. PMID- 10819047 TI - [Experimental study of combined effect of actoprotectors and radioprotectors in radiation injury]. AB - Combined treatment of mice and dogs with actoprotector bimethyl and radioprotector indralin suppressed radioprotective action of the latter, the surveillance being decreased from 83.3 to 14.3% in dogs and from 70.3 to 5% in mice. At the same time vasodynamic action of indralin (increase of artherial pressure up to 30% from a reference level) did not change. PMID- 10819048 TI - [The state of dog's metabolism after long interval after oxide tritium administration]. AB - An attempt to evaluate the influence of tritium oxide on the metabolism by some indices of lipid metabolism (common lipids, beta-lipoproteins, cholesterin), protein metabolism (cholinesterasa) and carbohydrate metabolism (blood sugar) was made. It was established that the introduction into organism of tritium oxide in the quantities, which could form lethal and sublethal doses of internal radiation, provoked the main changes of values of mentioned indices of metabolism. The character of metabolism changes in the remote period allows us to judge about the development of sclerosis processes, which can be the result of radiation-stipulated acceleration of organism aging. PMID- 10819049 TI - [Delayed biological effect of electromagnetic fields action]. AB - The real possibility of the development of remote effects in people after longterm EMF-exposure was presented in the world scientific literature. Many authors decided that there is connection between long-term EMF-exposure and development of the breast cancer, brain tumours, leukaemia and neurodegenerative diseases (Alzheimer's and Parkinson's diseases, amyotrophic lateral sclerosis). The analysis of up-to-date publications leads us to the conclusion that this problem is actual and further researches of conditions provoking development of last-term effects are required. PMID- 10819050 TI - [Cytogenetic and population effects in oligochaeta from the Chernobyl zone]. AB - Research on intensity and character of reproduction of Oligochaeta (Dero obtusa, Nais pseudobtusa and Nais pardalis) was carried out in 1995-1996 in two water bodies: a) in the Chernobyl NPP zone near Yanov Village, with a dose rate of 14 microGy h-1 on the surface of the bottom sediment; b) at the Uzh River area (the control water-body). The changes in both intensity and reproduction type were observed in worms at the radioactively polluted water-body. Stimulation of paratomic division in D. obtusa as well as activation of sexual reproduction in two other species were found out along with increasing of amount of cytogenetic damages in somatic cells. PMID- 10819051 TI - [Distribution and dynamic of 137Cs in arboreal plants]. AB - Dynamics of 137Cs specific activity (SA) in woody plants was examined. It was found that interseasonal variations of SA are connected mainly with precipitations and related oxygen in soil as well as Eh level. The rate of daily variation in SA in an individual tree can reach 19%. Acropetal or basipetal 137Cs distribution in the bole depends on physiological condition category (PCC) of a tree, weather and environment; SA ratio of higher and lower PCC depends on the same factors. In winter compared to summer higher SA correlation of tree separate parts was observed. Trees with 137Cs acropetal distribution in bark had proven lower general CA level. PMID- 10819052 TI - Adhesion molecules: clinical implications. PMID- 10819053 TI - Treatment of morbid obesity with the Swedish adjustable gastric band (SAGB): complication rate during a 12-month follow-up period. AB - BACKGROUND: The Swedish adjustable gastric band (SAGB) is used to treat morbid obesity. However, no quantitative data are available describing the follow-up of these patients with respect to the frequency and the complication rate of inflating and deflating the adjustable system. METHODS: We prospectively investigated 207 morbidly obese patients. All patients completed 12 months of follow-up and were seen in intervals of 1 to 3 months on an outpatient basis. RESULTS: A total of 207 patients had 1692 consultations (8.3 +/- 2.4 consultations per patient per year [mean +/- SD]), 920 port-a-cath punctions (4.6 +/- 2.0), 820 inflations (4.1 +/- 1.6), and 100 deflations (1.4 +/- 0.6). Complications related to the port-a-cath (n = 6, 2.9% of all patients) were 1 leakage of the tube (0.5%), 2 disconnections of the connecting tube (1.0%), and 3 reimplantations of the port-a-cath as a result of discomfort (1.4%). Complications related to the SAGB (n = 10, 4.8%) were 6 leakages of the band (2.9%), 2 penetrations (1.0%), 1 intraoperative perforation of the esophagus (0.5%), and 1 dystopically implanted SAGB (0.5%). Additionally, 9 minor early postoperative wound infections (4.3%) were recorded. CONCLUSIONS: Follow-up can be safely performed on an outpatient basis after implantation of a SAGB without infectious and other minor complications directly linked to the filling procedure. PMID- 10819054 TI - Invited commentary: laparoscopic adjustable gastric banding--a caution. PMID- 10819055 TI - Invited commentary: laparoscopic adjustable gastric banding--a viable option in bariatric surgery. PMID- 10819056 TI - The surgical treatment of isolated left-sided hepatolithiasis: a 22-year experience. AB - BACKGROUND: Various therapeutic methods are used in isolated left hepatolithiasis (ILH), and long-term results are not as satisfactory as they should be. METHODS: A retrospective analysis of 128 patients with ILH who were treated in our center over the last 22 years was undertaken to address patient age, gender, preoperative evaluation, operative findings, treatment modalities, and postoperative courses. RESULTS: Sixty of the 128 patients were male and 68 were female, with a mean age of 42 years (range, 16-72 years). Among these patients, most (81%) had stones in both left external hepatic duct (LEHD) and left medial hepatic duct; in contrast 13 patients (10%) had stones only in the LEHD, and 7 patients (6%) had stones only in the left hepatic duct. Concomitant strictures were present in the left hepatic duct, left medial hepatic duct, and LEHD in 60%, 76%, and 82%, respectively, most of which were severe. When compared with left lateral segmentectomy, left hepatic lobectomy had a lower rate of residual stones (4% vs 22%; P < .01). Residual/recurrent stones and ductal strictures were the 2 most common causes that affected the long-term results. Before June 1996, left lateral segmentectomy was performed more frequently than left hepatic lobectomy (59% vs 12%; P < .01); after June 1996, left hepatic lobectomy was performed more frequently (77% vs 13%; P < .01). Although there were no differences in length of operation, intraoperative bleeding, and postoperative complications, residual stones were more common after left lateral segmentectomy. CONCLUSIONS: Left hepatic lobectomy appears to be the most effective treatment for selected patients with ILH, if other operative procedures cannot remove all the related lesions, which include stones, dilation, stricture, or potential cholangiocarcinoma. PMID- 10819057 TI - Long-term survival and prognostic factors in the surgical treatment of mass forming type cholangiocarcinoma. AB - BACKGROUND: Mass-forming type cholangiocarcinoma is a distinct from of cholangiocellular carcinoma, with pathologic and biologic behavior different from those of other types. The clinical consequences of these differences have never been clarified. METHODS: Fifty-two consecutive patients (32 men and 20 women, mean age 62 years) with mass-forming type cholangiocarcinoma that had been treated with curative surgical resection between 1980 and 1998 were retrospectively evaluated. Long-term survival and disease-free survival were calculated, and univariate and multivariate analysis of various prognostic factors was conducted. RESULTS: The 30-day postoperative mortality rate was 2%, and the overall and disease-free 5-year survival rates were 36% and 34%, respectively. Univariate analysis identified 5 significant risk factors for overall survival: surgical margin, lymph node metastasis, lymph node dissection, vascular invasion, and left-side location of the main tumor. Two risk factors were identified for disease-free survival: surgical margin and lymph node metastasis. Multivariate analysis confirmed that surgical margin, lymph node metastasis, and vascular invasion were independently significant variables for overall survival. CONCLUSIONS: This is the first reported study on the effectiveness of liver resection for the treatment of mass-forming type cholangiocarcinoma, showing that surgical therapy can prolong survival if local radicality can be achieved and lymph-node metastases are absent. PMID- 10819058 TI - A subcutaneous or subfascial jejunostomy is beneficial in the surgical management of extrahepatic bile duct cancers. AB - BACKGROUND: Extrahepatic bile duct cancers are rare tumors with a dismal prognosis. Even after a resection, obstructive cholestasis and other biliary complications are the rule. To facilitate retrograde access to the biliary tree for treatment of such biliary complications, a modified Roux-en-Y hepaticojejunostomy is constructed such that the afferent limb is brought up as a subcutaneous or subfascial jejunostomy (SJ). The safety and utility of construction of an SJ was evaluated in patients with extrahepatic cholangiocarcinoma. METHODS: From 1985 to 1997, 24 patients with extrahepatic bile duct cancers received an SJ as part of their management. Demographic data, operative data, tumor characteristics, and postoperative courses were retrospectively reviewed. All but 3 patients were followed to the time of death. RESULTS: The average age of the patients was 62 +/- 9 years. The tumor was resected in 17 patients. Major complications occurred in 5 patients (21%). There was 1 operative death (4%). None of the complications could be attributed to construction of the SJ, although 1 patient had a soft tissue infection at the site of the percutaneous access of the SJ. Frequent dilatations of biliary strictures were required in 5 patients, and 1 patient eventually required insertion of an internal biliary stent. These procedures could all be accomplished through the SJ. CONCLUSIONS: The SJ is a technically simple and safe addition to the management of resectable and unresectable extrahepatic bile duct cancers, particularly proximal lesions. The procedure facilitates brachytherapy if indicated, and it allows convenient management of postoperative biliary complications, including recurrent strictures. PMID- 10819059 TI - Standardized measurement of the future liver remnant prior to extended liver resection: methodology and clinical associations. AB - BACKGROUND: There is no agreement regarding the preoperative measurement of liver volumes and the minimal safe size of the liver remnant after extended hepatectomy. METHODS: In 20 patients with hepatobiliary malignancy and no underlying chronic liver disease, volumetric measurements of the liver remnant (segments 2 and 3 +/- 1) were obtained before extended right lobectomy (right trisegmentectomy). The ratios of future liver remnant to total liver volume were calculated by using a formula based on body surface area. In 12 patients, response to preoperative right trisectoral portal vein embolization was evaluated. In 15 patients who underwent the planned resection, preoperative volumes were correlated with biochemical and clinical outcome parameters. RESULTS: The future liver remnants increased after portal vein embolization (26% versus 36%, P < .01). Smaller size liver remnants were associated with an increase in postoperative liver function tests (P < .05) and longer lengths of hospital stay (P < .02). Preliminary data indicates an increase in major complications for liver volumes < or = 25% (P = .02). CONCLUSIONS: A simple method of measurement provides an assessment of the liver remnant before resection. It is useful in evaluating response to portal vein embolization and in predicating the outcome before extended liver resections. PMID- 10819060 TI - Long-term experience after ex situ liver surgery. AB - BACKGROUND: Ex situ liver surgery allows liver resection and vascular reconstruction in patients who have liver tumors located at critical sites. Only a small series of studies about ex situ liver surgery is available in the literature. No long-term results have been published. METHODS: Twenty-four patients were considered for ex situ liver surgery because conventional liver surgery was considered impossible or too hazardous. The patient's ages were 51.3 +/- 7.5 years. Indications were various primary and secondary liver malignancies and benign liver tumors in 2 patients. RESULTS: In 22 of 24 patients, the ex situ liver resection and subsequent autotransplantation were performed. The anhepatic periods in these patients lasted for 5.6 +/- 1.1 hours. In the remaining 2 patients, autotransplantation was not possible and allogenic liver transplantation was performed 17 and 19 hours after hepatectomy. In 4 patients, liver failure occurred after autotransplantation and required transplantation. The confluens between hepatic veins and the inferior vena cava was reconstructed in 5 patients. Fifteen patients survived the postoperative period and were discharged after 36.5 +/- 16 days. The median survival time of 6 patients who had metastases of colonic carcinoma was 21 months. The 2 patients with benign liver disease are alive 9 and 5 years after ex situ surgery. CONCLUSIONS: Extended liver resections with difficult reconstructions of the hepatic venous confluens are feasible by ex situ liver surgery and subsequent autotransplantation. However, the early postoperative mortality rate is high, especially in patients with cholestatic livers. Early tumor recurrence remained the problem in these patients with extended local tumor spread. Ex situ liver surgery should only be performed in selected patients. PMID- 10819061 TI - Aggressive surgical approach to recurrent tumors after hepatectomy for metastatic spread of colorectal cancer to the liver. AB - BACKGROUND: Liver resection is currently accepted as the only potential cure for patients with metastases of colorectal tumors in the liver. However, cancer will recur in more than 70% of patients. METHODS: In the 7 years to December 1997, 60 patients underwent liver resections for colorectal metastases at our institute. Of these, 20 patients had repeated surgical resections for recurrent disease of the liver and other organs. Another 2 patients had undergone previous hepatectomy elsewhere. The clinical data for these patients were reviewed. RESULTS: The median interval between the 2 resections was 16 months. Eighteen hepatectomies, 6 lung resections, and 1 pancreatoduodenectomy were performed in 22 patients. Operative mortality and complication rates were 0% and 18%, respectively. At a median follow-up of 25 months after repeated resection, the survival rate in these patients was 73% at 2 years (12 of 16 evaluable patients are surviving) and 22% at 5 years (2 of 10 evaluable patients are surviving); the median survival time was 44 months. CONCLUSIONS: Repeated resections for recurrent colorectal metastases yield comparable results to first liver resections in operative mortality and morbidity rates, survival time, and pattern of recurrence. Although the number of patients surviving more than 5 years is still limited, the absence of other proven treatments supports the concept of an aggressive resectional approach for these patients. PMID- 10819063 TI - Prolonged preservation increases surgical complications after pancreas transplants. AB - BACKGROUND: Prolonged preservation of the donor organ may result in delayed graft function or nonfunction after most organ transplants. We studied whether or not prolonged preservation increases surgical complications after pancreas transplants. METHODS: Between January 1, 1994, and September 30, 1998, a total of 294 recipients underwent cadaver pancreas transplants at our institution. Recipients were analyzed in 2 groups: those with pancreas preservation time < or = 20 hours (n = 211) versus > 20 hours (n = 83). RESULTS: Demographic data were similar between the 2 groups, except that mean donor age in the prolonged preservation group was significantly lower. Despite use of younger donors, prolonged preservation was associated with an increased incidence of surgical complications, most notably leaks, thrombosis, and wound infections. Grafts with prolonged preservation were more often noted by the transplant surgeon to be edematous after reperfusion, although the incidence of hyperamylasemia posttransplant did not differ between the 2 groups. Graft and patient survival rates also did not differ between the 2 groups. The incidence of early graft loss (< 3 months) was, however, higher in the prolonged preservation group (20.5% versus 9.0%, P = .04). CONCLUSIONS: Prolonged preservation of the donor organ increases the incidence of surgical complications after pancreas transplants. All attempts should be made to minimize preservation time, keeping it below 20 hours, if possible. PMID- 10819062 TI - Intraductal papillary mucinous tumors of the pancreas: pancreatic resections guided by preoperative morphological assessment and intraoperative frozen section examination. AB - BACKGROUND: We undertook this study to assess the results of a policy of limited pancreatic resections for intraductal papillary-mucinous tumors (IPMT), guided by a standardized preoperative morphological assessment and the frozen section histologic examination of pancreatic resection margins. METHODS: From 1991 to 1998, there were 41 patients who underwent resection of IPMT in our center after standardized morphological preoperative assessment, including abdominal computed tomography scans, endoscopic retrograde cholangiopancreatography, and endoscopic ultrasonography. All patients but one underwent a partial pancreatic resection. RESULTS: Preoperative assessment had an 89% diagnostic accuracy for IPMT. It detected malignant transformation with a 67% sensitivity and a 95% specificity, but failed to correctly assess ductal extension of the disease in 17 cases (41%). The diagnostic accuracy of frozen section histologic examination of the pancreatic resection margin was 92%. There was no recurrence in case of in situ carcinoma when epithelial lesions were completely resected (8 cases) nor in lower grades of dysplasia (23 cases). Conversely, recurrence was constant in invasive carcinoma (10 cases) regardless of the status of the pancreatic margin. CONCLUSIONS: Frozen section histologic examination of the pancreatic resection margin is useful and reliable to ensure a complete resection of IPMT by a partial resection that prevents recurrence of noninvasive lesions. PMID- 10819064 TI - Clinical application of malignancy potential grading as a prognostic factor of human esophageal cancers. AB - BACKGROUND: Various biologic markers have been reported to be prognostic factors in human esophageal cancers. In the current study, we established a new tumor grading system representing the malignancy potential of cancer cells and compared it with the clinical-stage system. METHODS: Tumor samples from 77 patients with squamous cell carcinoma of the esophagus were immunohistochemically evaluated for the expression of 10 molecules: the cell cycle-related molecules of cyclin D1, Rb, p16INK4, p27KIP1, and PCNA; the cell-cell adhesion molecules of E-cadherin, alpha-catenin, and beta-catenin; and the heat shock proteins of HSP27 and HSP70. RESULTS: P27KIP1, beta-catenin, and HSP70 were selected for their high hazard ratio in multivariate analysis, and the number of their disordered molecules was used to define the malignancy grade (MG). Five-year survival rates were 83%, 54%, 17%, and 0% for MG1, MG2, MG3, and MG4. The gradation of survival curves was better for MGs than for clinical stages. MGs and clinical stages showed significant correlation; however, 55% of those in higher clinical stages (stage 3 or 4) had lower MG (MG1 or 2) and showed better prognosis than others in their group (stage 3 or 4 and MG3 or 4). The proportions of shorter survival span to cancer death patients (less than 1 year) were 0%, 33%, 75%, and 100% in MG1, 2, 3, and 4, but the clinical stage was not associated with the survival span. CONCLUSIONS: The grading of malignancy potential is clinically useful, especially for selecting patients who may show good prognosis in the advanced clinical stage and for predicting short survival span. These predictions are not possible with the clinical-stage system, which is based on the anatomic spread of cancer cells. PMID- 10819065 TI - Reoperation for secondary uremic hyperparathyroidism: are technical difficulties influenced by initial surgical procedure? AB - BACKGROUND: Parathyroid surgery in patients with uremia and secondary hyperparathyroidism is performed either by subtotal parathyroidectomy or total parathyroidectomy with immediate reimplantation. The aim of this study was to compare the results of reoperation for persistent or recurrent hyperparathyroidism after parathyroidectomy according to which initial operative procedure was used. PATIENTS AND METHODS: Eighty-nine patients had reoperation for persistent (28 patients) or recurrent (61 patients) hyperparathyroidism after 53 subtotal parathyroidectomies and 36 total parathyroidectomies with immediate reimplantation. Results of the reoperation were assessed in terms of success rate, morbidity, and operative findings. RESULTS: The success rate of reoperation in patients with persistent hyperparathyroidism was 89% and was independent of the initial type of surgery. Success rates of reoperation for recurrent hyperparathyroidism after initial subtotal parathyroidectomy and total parathyroidectomy with immediate reimplantation were 87% and 70%, respectively (P = .02). Hypertrophy of the parathyroid remnant was the main cause of recurrence after subtotal parathyroidectomy. After total parathyroidectomy with immediate reimplantation, recurrence was located in the graft in half the patients, while hyperplastic tissue was found in the neck or the mediastinum in the other half. CONCLUSIONS: Subtotal parathyroidectomy provides the best conditions for successful reoperation in case of recurrent hyperparathyroidism and should become the surgical treatment of choice for secondary hyperparathyroidism. PMID- 10819066 TI - Carbon dioxide pneumoperitoneum is associated with increased liver metastases in a rat model. AB - BACKGROUND: In patients with primary colorectal cancer, liver metastases are the most important indicators of prognosis and survival; the effect of laparoscopic surgery on the development of hepatic tumor spread is still largely unknown. METHODS: Thirty WAG/Rij rats were randomly divided into 3 operative groups for intraportal tumor cell inoculation: carbon dioxide pneumoperitoneum (group I, n = 10), gasless laparoscopy (group II, n = 10) and open laparotomy (group III, n = 10). The total operating time was 90 minutes with tumor cell injection (50,000 CC531 colon carcinoma cells/mL) performed 45 minutes after the start of the procedure. Hepatic tumor growth and the total tumor load were evaluated 28 days after surgery. RESULTS: Hepatic tumor growth and total tumor load were significantly reduced in the gasless laparoscopy group (group II) as compared with the carbon dioxide pneumoperitoneum group (group I) and the open laparotomy group (group III) (P < .05). No significant difference was found between the carbon dioxide and the open laparotomy groups. CONCLUSIONS: Insufflation of carbon dioxide may actually stimulate metastatic disease of the liver. Gasless laparoscopy seems to preserve hepatic resistance against tumor growth. PMID- 10819067 TI - A new method of intrathecal PO2, PCO2, and pH measurements for continuous monitoring of spinal cord ischemia during thoracic aortic clamping in pigs. AB - BACKGROUND: Impaired spinal cord circulation during thoracic aortic clamping may result in paraplegia. Reliable and fast responding methods for intraoperative monitoring are needed to facilitate the evaluation of protective measures and efficiency of revascularization. METHODS: In 11 pigs, a multiparameter PO2, PCO2, and pH sensor (Paratrend 7, Biomedical Sensors Ltd, United Kingdom) was introduced into the intrathecal space for continuous monitoring of cerebrospinal fluid (CSF) oxygenation during thoracic aortic cross-clamping (AXC) distal to the left subclavian artery. A laser-Doppler probe was inserted into the epidural space for simultaneous measurements of spinal cord flux. Registrations were made before and 30 minutes after clamping and 30 and 60 minutes after declamping. The same measuring points were used for systemic hemodynamic and metabolic data acquisition. RESULTS: The mean CSF PO2 readings of 41 mm Hg (5.5 kPa) at baseline decreased within 3 minutes to 5 mm Hg (0.7 kPa) during AXC (P < .01). Spinal cord flux measurement responded immediately in the same way to AXC. Both methods indicated normalization of circulation during declamping. Significant (P < .01) changes were also observed in the CSF metabolic parameters PCO2 and pH. CONCLUSIONS: In this experimental model of spinal ischemia by AXC, online monitoring of intrathecal PO2, PCO2, and pH showed significant changes and correlated well with epidural laser-Doppler flowmetry (P < .01). PMID- 10819068 TI - Effect of endothelial and adventitial injury on somatostatin receptor expression. AB - BACKGROUND: The somatostatin analog, angiopeptin, inhibits intimal hyperplasia formation; although the specific somatostatin receptor (SSTR) subtypes transducing this effect are unknown. The purpose of this study was to determine the expression of SSTR subtypes in rat iliac arteries after balloon catheter endothelial injury and perivascular dissection. METHODS: Male rats received balloon endothelial injury to their left common and external iliac arteries with or without circumferential arterial dissection. The right arteries served as controls. At 1 and 2 months after intimal injury, animals were killed and their iliac arteries harvested and studied for SSTR expression by using immunocytochemical and molecular techniques. Quantitative polymerase chain reaction was used to determine the level of SSTR expression. RESULTS: Normal rat iliac arteries expressed only SSTR2 and 3. After balloon endothelial injury, there was significant upregulation of SSTR2 messenger RNA at 1 and 2 months after injury as compared with controls (1 month, 1.8 +/- 0.3 vs 0.4 +/- 0.1 zmol, P < .001; 2 months, 2.7 +/- 0.5 vs 1.1 +/- 0.2 zmol, P < .001). The addition of adventitial dissection to endothelial injury also showed a significant increase in SSTR2 expression (1 month, 2.4 +/- 0.4 vs 0.8 +/- 0.2, P < .05; 2 months, 1.3 +/- 0.3 vs 0.7 +/- 0.3, P < .05), but not significantly greater than that seen after balloon endothelial injury alone. Immunocyto-chemical studies also demonstrated an increase in SSTR2 immunoreactivity on the luminal surface of the endothelial cells in the balloon catheter-injured arteries. CONCLUSIONS: These findings show that SSTR2 is the primary SSTR that is upregulated after injury and likely mediates the effects of somatostatin analogs on intimal hyperplasia. PMID- 10819069 TI - Lactating adenoma presenting as a giant breast mass. PMID- 10819070 TI - Recombinant granulocyte macrophage colony stimulating factor (rhu-GM-CSF) in the treatment of extensive leg ulcers: a case report. PMID- 10819071 TI - Ulcer necessitans: a forgotten entity. PMID- 10819072 TI - Factor V Leiden mutation in a patient with warfarin-associated skin necrosis. PMID- 10819073 TI - Early work in lymphatic mapping. PMID- 10819074 TI - Endoscopic sphincterotomy in the management of anomalous pancreaticobiliary junction. PMID- 10819075 TI - Glycoproteins: rapid sequencing technology for N-linked and GPI anchor glycans. PMID- 10819076 TI - Biomedical and pharmaceutical applications of alginate and chitosan. PMID- 10819077 TI - Biopolymer mucoadhesives. PMID- 10819078 TI - Biotechnological strategies for the modification of food lipids. PMID- 10819079 TI - Polysialic acids: potential role in therapeutic constructs. PMID- 10819080 TI - Microbial polysaccharide products. PMID- 10819081 TI - The production and applications of genetically modified skin cells. PMID- 10819082 TI - The functions of 4-alpha-glucanotransferases and their use for the production of cyclic glucans. PMID- 10819083 TI - Thermomechanical properties of amorphous saccharides: their role in enhancing pharmaceutical product stability. PMID- 10819084 TI - Xylans of industrial and biomedical importance. PMID- 10819085 TI - Pectins, pectinases and plant-microbe interactions. PMID- 10819086 TI - The unfolding story of the chaperonins. PMID- 10819087 TI - Overview of neuropathic pain syndromes. AB - Patients with peripheral nerve injury often express a variety of positive sensory symptoms. Description of these symptoms can range from "painful" to "painless" to "painless but very unpleasant." Although it is widely accepted that both peripheral and central processes may play a role in the pathophysiology of these symptoms, peripheral mechanisms in particular are often overlooked. Sensitization of nociceptor unit receptors, spontaneous or stimuli-induced ectopic impulse generation, "multiplication" of impulses, and ephaptic transmission are pathophysiologic mechanisms resolved at the peripheral level that may explain the patient's symptomatology. It is important to reach an accurate diagnosis and to investigate the pathophysiology of every individual symptom. Only when this has been achieved is it possible to plan adequate symptom-oriented therapeutic strategies. In the future, therapy will be more symptom-oriented than illness oriented, and treatment options will most likely be tailored to fit the specific constellation of symptoms expressed by a particular patient. PMID- 10819088 TI - Mechanisms of pain in peripheral neuropathy. AB - Over the last few years, the mechanisms of pain due to peripheral nerve injury have been the subject of extensive clinical and fundamental investigation. Several types of peripheral mechanisms have been described in animal models of peripheral nerve injury. Abnormal (ectopic) neuronal activity has been reported in primary afferents and in the dorsal root ganglion, and appears related to dysregulation of the synthesis and/or the functioning of sodium channels (notably the tetrodotoxin-resistant channel). Fiber interactions (ephaptic or cross excitation), nociceptor sensitization and sympathetic sensory coupling may also be involved in some cases. Peripheral nerve lesions can also induce central changes; this has essentially been investigated at the spinal cord level in animals. Three major types of modifications could induce a pathologic activation of central nociceptive neurons: modification of the modulatory controls of the transmission of nociceptive messages; anatomic reorganization (neuroplasticity) of the central nociceptive neurons, and thus their pathologic activation; and central sensitization (hyperexcitability) of nociceptive neurons to produce modifications of their electrophysiologic properties. Central sensitization probably depends critically on intracellular changes induced by the activation of N-methyl-D-aspartate (NMDA) receptors by excitatory amino acids released by primary afferents. Due to the multiplicity of mechanisms, it is unlikely that neuropathic pain corresponds to a unique entity. Each of the painful symptoms may correspond to distinct mechanisms and thus respond to specific treatments. PMID- 10819089 TI - Treatment options in postherpetic neuralgia. AB - Postherpetic neuralgia (PHN) is a separate disease entity that represents a complication of acute herpes zoster. PHN, involving aberrant somatosensory processing in the peripheral and/or central nervous system, is considered to be a chronic neuropathic pain, frequently unresponsive to all treatment modalities. Despite the clinical trial data demonstrating successful pain relief with several drug regimens, the pharmacologic management of neuropathic pain is difficult, particularly in PHN. Response to therapy is generally inhomogeneous. Some patients experience long-term pain control with either topical or oral monotherapy with antidepressants, anticonvulsants, or opioids. Other PHN patients, such as those suffering pain due to central nervous system lesions, are extraordinarily refractory to all measures. This article will review current treatments--tricyclic antidepressants, anticonvulsants, local anesthetics, clonidine, N-methyl-D-aspartate (NMDA)-antagonists, and opioids and focus on mechanism-based pharmacologic interventions. Pharmacologic approaches can be classified into three groups: 1) drugs that act topically in the affected skin area; 2) drugs that act on nerve excitability and conduction in sensory axons; and 3) drugs that act on neural damage related synaptic changes. This last group is the only pain treatment option related to central denervation. To date, the treatment of PHN has relied on the use of tricyclic antidepressants (TCAs), which represent the most comprehensively studied medications for this pain syndrome. Clinical data indicate that TCAs are effective analgesics in approximately 50% of patients; these drugs have been recommended as first-line agents for all neuropathic pain syndromes except trigeminal neuralgia, but are frequently contraindicated or poorly tolerated in elderly patients with PHN. If monotherapy fails, a mechanism- and/or symptom-based multidrug regimen can be used. There is also consistent support for intravenous and topical lidocaine, intravenous ketamine, carbamazepine, and opioids. Gabapentin, a new anticonvulsant, can be considered a first-line oral medication for PHN based on the efficacy and safety results of a recently completed double-blind trial. In addition to positive effects on PHN, sleep, mood, and overall quality of life were significantly improved. PMID- 10819090 TI - Treatment options in painful diabetic neuropathy. AB - Diabetic neuropathy is common in patients with diabetes mellitus, and 7.5% of diabetics experience pain from diabetic neuropathy. Complications of diabetes mellitus are more common where control of the disease is not optimal. By improving the control of the disease, both the neuropathy and the pain it can produce may be improved. The pain of diabetic neuropathy can frequently be controlled using analgesics, antidepressants, anticonvulsants, topical capsaicin, and neuromodulation, either alone or in any combination. PMID- 10819091 TI - Gabapentin: a new tool in the treatment of neuropathic pain. AB - Human neuropathic pain remains a prevalent and pervasive problem in our society. Pharmacologically there is also no single, uniformly well-tolerated drug that is reliably helpful. Gabapentin has emerged as a useful new tool based on the results of two large multicenter trials in models of human neuropathy. Gabapentin proved to be a significantly better analgesic than placebo, was well tolerated in the elderly population, and had a significant positive impact on several subjective and objective outcome measures. A discussion of the standard treatments and the studies supporting this new tool is the purpose of this review. PMID- 10819092 TI - Failure-to-thrive, maltreatment and the behavior and development of 6-year-old children from low-income, urban families: a cumulative risk model. AB - OBJECTIVE: A cumulative risk model was used to examine the relationship among failure-to-thrive (FTT), maltreatment, and four aspects of children's development: cognitive performance (standardized testing), adaptive functioning at school, and classroom behavior (teacher report), and behavior at home (maternal report). METHOD: The sample included 193 6-year-old children and their families, recruited from pediatric clinics serving inner-city, low-income, primarily African-American families, who were part of a longitudinal investigation of child development and maltreatment. Four risk groups were formed based on their growth and maltreatment history: neither FTT nor Maltreatment, FTT Only, Maltreatment Only, and both FTT and Maltreatment. FTT was defined as a deceleration in weight gain (weight-for-age below the 5th percentile) prior to 25 months of age among children born at term with birth weight appropriate for gestational age. Maltreatment was defined as having at least one report to CPS for neglect, physical abuse and/or sexual abuse. RESULTS: Risk status was negatively associated with each of the four developmental outcomes. Children with a history of both FTT and maltreatment had more behavior problems and worse cognitive performance and school functioning than children with neither risk factor. Children with only one risk factor (either FTT or maltreatment) achieved intermediate scores. CONCLUSIONS: Findings support a cumulative risk model as being more detrimental to children's development than the presence of a single risk factor alone, consistent with theories linking the accumulation of environmental risks to negative consequences. These results underscore the importance of interventions to prevent both FTT and maltreatment during children's early years. PMID- 10819093 TI - What progress has been made in meeting the needs of seriously maltreated children? The course of 200 cases through the Boston Juvenile Court. AB - OBJECTIVES: The study examined child, parent, and case characteristics in a sample of 200 cases of serious child maltreatment brought before the Boston Juvenile Court (BJC) on Care and Protection petitions in 1994. Whether recent changes in Massachusetts law have been effective in reducing delays in adjudication and helping children achieve permanent placements more quickly was also examined. METHOD: Data were abstracted from court records by the research team. The 200 cases were followed prospectively for 4 years. Retrospective data on the families' previous involvement with the protective service system were also abstracted from the records. Data from the 1994 cases were compared to that obtained from a sample of cases brought before the BJC in 1985-1986. RESULTS: Children permanently removed from parental custody in the 1994 sample required less time post-disposition to achieve permanent placements. However, overall, time frames for the 1994 cases remained remarkably similar to those in 1985-1986: children were in the protective service system an average of 5 years; cases required an average of 1.6 years in court; and half of the children permanently removed from parental custody were still in "temporary" foster care at 4-year follow-up. CONCLUSIONS: Although some improvements have occurred since 1985-1986, the system still fails to meet the needs of seriously maltreated children to achieve permanent placements promptly. The implications of the findings for system reform are discussed. PMID- 10819094 TI - Psychosocial correlates of child abuse potential in multiply disadvantaged adolescent mothers. AB - OBJECTIVE: This study had two aims: First to examine psychosocial correlates of child maltreatment risk, and second to assess the validity of the CAP Inventory (Milner, 1986) with multiply disadvantaged teenage mothers. METHOD: Participants were 75 adolescent mothers who were wards of the Illinois child protection system. Mothers (aged 14-18) and infants participated in home-based psychosocial assessment of personal and parenting functioning. Group comparisons examined differences for mothers with elevated versus normal versus invalid CAP scores due to faking good. RESULTS: Findings indicated that abuse risk groups differed on emotional distress, social support satisfaction, reading achievement, and years of education, but not on parenting beliefs or quality of child stimulation. Differences favored the normal over the elevated risk group in all significant comparisons, whereas mothers with elevated faking good differed from normals only in lower reading achievement. Multiple regression analysis highlighted emotional distress, support dissatisfaction, and low achievement as significant predictors of greater abuse risk. CONCLUSIONS: Despite sharing multiple disadvantages, adolescent wards are a heterogeneous group who show different levels of psychosocial functioning corresponding to levels of child maltreatment risk. The findings provide support for the concurrent validity and clinical applicability of the CAP Inventory with disadvantaged teenage mothers. PMID- 10819096 TI - The relationship of social support to physically abused children's adjustment. AB - OBJECTIVE: This study had three main objectives: First, to assess physically abused children's perceptions of teacher, peer, and family support; second, to determine whether the levels of perceived support differ according to the person's social role; and third to assess which sources of social support show stronger associations with adjustment in a physically abused sample. METHOD: Perceived social support from teachers, families and peers was assessed in a sample of 37 physically abused children using a shortened version of the Survey of Children's Social Support (Dubow & Ullman, 1989). Child adjustment was indexed by child and parent reports of child depression, anxiety, and anger. RESULTS: Analyses indicated that the children rated their families, peers, and teachers highly as sources of social support, with families being rated as the most important source. Hierarchical multiple regression analyses indicated that perceived peer support was significantly negatively related to children's and parent's reports of children's depression and anxiety. Furthermore, perceived family support was significantly negatively associated with child reported depression. No significant relationships were found between perceived teacher support and symptomatology. CONCLUSIONS: Overall, the results suggest that peer and family support are particularly important for physically abused children's psychological functioning, particularly for internalizing problems. PMID- 10819095 TI - The abused child as parent: the structure and content of physically abused mothers' perceptions of their babies. AB - OBJECTIVE: The major aim of the study was to provide an empirical answer to the following question: Does a mother's history of being physically abused as a child have a discernible impact on the structure and content of her perceptions and beliefs concerning her own child? METHOD: Free-response memories and current descriptions of babies, self, and significant others such as parents were compared longitudinally in two groups of mothers when their babies were 6 months, 1 year, and 2 years old. One group of mothers consisted of individuals who reported being physically abused as children; the control group consisted of mothers who were not physically abused. The two groups were comparable with respect to age of baby, race, and socioeconomic status. RESULTS: Abused mothers were found to differ significantly from control mothers in the structure and content of their free-response perceptions of their own babies. More specifically, abused mothers lagged behind controls in how well-differentiated were their negative perceptions of their babies. Differentiation in this study is operationally defined as the number of unique clusters that underlie a mother's perceptions of her baby, when social perception data is analyzed using cluster analysis (HICLAS). The greater the number of clusters observed, the greater is the differentiation. On the other hand, abused mothers were comparable to controls with respect to differentiation of positive perceptions of babies. CONCLUSIONS: The findings constitute a discovery about the structural organization of social cognition in mothers at-risk for child abuse. Implications of the findings for theory and future research are briefly discussed, as are limitations of the current study. PMID- 10819097 TI - Dissociation as a mediator between child abuse history and adult abuse potential. AB - OBJECTIVE: It has commonly been found that abused children are at risk for later becoming abusive parents (Kaufman & Zigler. 1987; Oliver, 1993) and observational learning has been discussed as a mechanism that perpetuates this intergenerational cycle of abuse. However, two thirds of abused children do not become abusive (Kaufman & Zigler, 1987). Thus, the goal of the current study was to examine whether dissociation functions as an additional mechanism mediating the relation between a history of child abuse and abusiveness as an adult. METHOD: A cross-sectional design was used to examine physical abuse history, dissociation, and physical abuse potential in a sample of 190 college students. Questionnaires were utilized to assess the three constructs. RESULTS: Findings indicated that the three constructs were intercorrelated. In particular, a newly established association was found between dissociation and physical abuse potential (r = .54, p < .0001). The primary finding was that the relation between physical abuse history and physical abuse potential was significantly mediated (z = 2.19, p < .05) by level of dissociation, with dissociation accounting for approximately half of the observed relation between history of abuse and abuse potential. CONCLUSION: Results suggested that dissociation may be one mechanism that helps to perpetuate the intergenerational cycle of abuse. Although dissociation promotes psychological survival during children's abuse experiences, it may result in the development of abusive tendencies in later life. Reducing parental dissociation may assist clinicians in preventing or terminating physical child abuse. PMID- 10819098 TI - The prevalence of a history of child sexual abuse among adults visiting family practitioners in Israel. AB - OBJECTIVE: To determine the prevalence of a history of child sexual abuse (CSA) in a random sample of adult patients presenting for routine health care to family practice clinics in Israel. METHOD: One thousand and five randomly selected patients aged 18 to 55, attending 48 clinics, participated in this questionnaire study. RESULTS: Twenty-five percent indicated that they had been sexually abused as children. More women reported CSA (p < .0001 ) than men, as did women originating from Western countries (p = .02) and those with more than 12 years of education (p = .01). There were no significant associations between CSA and the other socio-demographic variables examined. Fondling was the most common and intercourse the least common activity experienced. Forty-five percent of the perpetrators were previously known. The mean age at which the child sexual abuse began varied between 10 and 14. Only 45% of the subjects had ever told anyone about the experience. CONCLUSIONS: Since no other prevalence study has been reported to date in Israel, these findings suggest that as in other Western countries CSA is a relatively common problem. Family physicians and other health professionals should be aware of this high prevalence and its known potential for initial and long-term deleterious outcomes. PMID- 10819099 TI - Assessing the relationship between child sexual abuse and marginal living conditions on HIV/AIDS-related risk behavior among women prisoners. AB - OBJECTIVE: There were two aims in this research. First, to examine the relationships between childhood sexual abuse and HIV drug and sexual risk taking behaviors among female prisoners, and second, to examine the relationship between a marginal adult living context and HIV drug and sexual risk taking behavior among female prisoners. METHOD: The data were collected through face-to-face interviews with a random sample of 500 women at admission to prison in 1994. Differences between women who were sexually abused while growing up (n = 130) were compared to women who reported no sexual abuse (n = 370) along various demographic, and HIV drug and sexual risk taking dimensions. RESULTS: A history of sexual abuse while growing up was associated with increased sexual risk taking behaviors in adulthood. A marginal adult living situation also emerged as an important factor increasing the risk for HIV infection. Examining the co occurrence of both childhood sexual abuse and adult marginal living context revealed a strong relationship between these two factors and HIV risk taking activities. CONCLUSIONS: The findings indicate that childhood sexual abuse may be a predictor for HIV sexual risk taking behaviors among incarcerated women. The marginal and chaotic adult living style of these women was also associated the extent of their HIV drug and sexual risk taking behaviors. Our research suggests that the co-occurrence of sexual victimization and marginality is a stronger predictor of HIV risk than each variable alone. PMID- 10819100 TI - Criminal investigation of child sexual abuse: a comparison of cases referred to the prosecutor to those not referred. AB - OBJECTIVE: The present study sought to identify characteristics of child sexual abuse cases which differentiate cases referred for criminal prosecution ("criminal-action") from those not referred ("dropped") by investigators. METHOD: The study sample consisted of 1043 children who completed a forensic interview for sexual abuse that allegedly occurred at the hands of an adult between January 1, 1993 and December 31, 1996 in Bernalillo County of New Mexico. Data was systematically obtained from forensic interview files and offender records at the local prosecutor's office. RESULTS: Differences between criminal-action and dropped cases were found in relation to the children (age, sex and ethnicity), the alleged offenders (age, sex and relationship to child), and the case characteristics (disclosure and injury to the child). CONCLUSIONS: The present study provided insight into the characteristics of a previously ignored population (reported child sexual abuse cases that are not referred for prosecution). Recommendations are made to address the needs of these children and their families. PMID- 10819101 TI - Childhood history of abuse and child abuse potential in adolescent mothers: a longitudinal study. AB - OBJECTIVES: The purpose of this research was to determine whether adolescent mothers of newborns are at higher risk for child abuse than adult mothers of newborns and to examine whether adolescent mothers with memories of child maltreatment have a higher risk for child abuse. METHOD: Two groups (adolescents and adults) of pregnant mothers were followed for 20 months beginning between the 5th and the 7th month of pregnancy until the child was 18 months old. Adolescent (N = 24) and adult (N = 24) mothers were matched on sociodemographic variables. During pregnancy, memories of child maltreatment were evaluated. When child was 1, 6, 12, and 18 months old, risk for child abuse was evaluated. RESULTS: Adolescent and adult mothers showed no differences in memories of childhood physical or emotional abuse. Nevertheless, adolescent mothers showed higher child abuse potential and depression scores than adult mothers. Mothers with memories of severe physical punishment showed higher child abuse potential scores and mothers with memories of physical punishment producing physical damage showed higher child abuse potential and depression scores. A statistically significant age of the mother by physical punishment producing physical damage interaction was found for depression. CONCLUSIONS: The results of this longitudinal study indicated that the potential for abuse was significantly greater in adolescent mothers than in adult mothers, and in mothers who had been victims of physical abuse than in those who had not. It also appeared that, among adolescent mothers, those who had been victims of childhood physical abuse constitute a higher risk group for child physical abuse. PMID- 10819102 TI - A history of abuse and negative life events in patients with a sexually transmitted disease and in a community sample. AB - OBJECTIVE: Our aim was to investigate the psychological impact of abuse and negative life events during childhood, adolescence and adulthood in patients recruited from a sexual health clinic. METHOD: Sixty-two patients with a sexually transmitted disease (STD) were matched on age and gender with a community sample. Forty-two patients without an STD formed another control group. Subjects self reported their trauma histories and current psychological distress. RESULTS: Compared to controls, categories of abuse and negative life events were more prevalent in STD patients, particularly Physical/Sexual Abuse in adolescence and adulthood. Both within the STD and community samples, a negative life event category that measured illness/death of loved ones during childhood and adolescence predicted current psychological distress. In addition, Control Abuse (a subtype of psychological abuse involving selfish manipulation and deprivation) in childhood and adolescence strongly predicted current psychological distress in STD patients. In general, effects were stronger in females than in males. Psychological/Verbal Abuse did not independently predict current psychological distress, but accompanied other abuse types and possibly amplified their adverse effects. CONCLUSIONS: Our findings suggest that multiple types of abuse and negative life events increase the risk of STD infection, perhaps by increasing the likelihood of multiple sexual partners. These patients may ignore social conventions of sexual behaviour because they are bitter about past life experiences. Alternatively, they may persistently search for affection to compensate for a lack of affection in the past. PMID- 10819103 TI - Management and outcome of glottic cancer: a population-based comparison between Ontario, Canada and the SEER areas of the United States. Surveillance, Epidemiology and End Results. AB - OBJECTIVE: We compared treatment practice and outcome in glottic cancer in Ontario, Canada to that in the Surveillance, Epidemiology and End Results (SEER) program areas in the United States to determine whether the Ontario emphasis on the use of delayed combined therapy was associated with similar survival and better laryngectomy-free survival than the U.S. approach, which emphasizes greater use of surgery. METHODS: Electronic, clinical, and hospital data were linked to cancer registry data. The study groups compared on survival comprised all patients diagnosed from 1982 to the end of 1991 in Ontario (2324 patients) and in the SEER areas (5715 patients). Comparisons on initial treatment, laryngectomy rates, and laryngectomy-free survival were limited to subsets of these study populations due to data availability. Initial treatment data were provided by the SEER registries in the U.S. and by the cancer clinic and hospitalization data in Ontario. Information about laryngectomies performed subsequent to initial treatment was available from Medicare hospitalization data in the U.S. and from Canadian Institute for Health Information hospitalization data in Ontario. RESULTS: Although radiotherapy was the most common initial treatment in both areas, it was used more often in Ontario (84.4% versus 63.2% in the U.S. [p < 0.001]). Relative survival was not statistically different with a relative risk comparing SEER to Ontario of 1.09, 95% confidence interval (CI) (0.93, 1.29). Laryngectomy rates were similar with a relative risk of 1.01, 95% CI (0.67, 1.52), and it follows from the survival and laryngectomy rate comparisons that the laryngectomy-free survival was not statistically different (p = .95). CONCLUSIONS: There are large differences in the management of glottic cancer between the U.S. and Ontario and no corresponding differences in survival or laryngectomy-free survival. This work highlights a need for more clinical investigation into the relative merits of differing management policies in glottic cancer. PMID- 10819104 TI - Laser facial skin resurfacing: discussion on erbium:YAG and CO2 lasers. AB - There are two lasers for resurfacing. The erbium:YAG laser and the CO2 laser both have different physical and physiologic differences when striking the skin. Therefore, each laser gives a different result. This article reviews the use of the Sharplan Silktouch CO2 laser for 4 years and the erbium:YAG laser for 1 year. In the last year, we have performed 40 laser procedures with the erbium:YAG laser. Discussion ensues regarding the differences and similarities of the two lasers as well as the difficulties and complications. PMID- 10819105 TI - Does intratympanic gentamicin treatment for Meniere's disease cause complete vestibular ablation? AB - OBJECTIVE: To determine changes in vestibular function following intratympanic gentamicin (ITG) treatment for Meniere's disease and to correlate changes with the need for further treatment. STUDY DESIGN: Prospective case series. PATIENTS: One hundred and three patients with disabling unilateral Meniere's disease who had failed a minimum of 6 months medical treatment. MAIN OUTCOME MEASURES: Vestibular function was measured by electronystagmography (ENG) caloric testing before and serially following treatment. Caloric responses were classified as normal (excitability difference [ED] < 25%), bithermal response (ED > or = 25%), positive response to ice water only, and absent ice water response. RESULTS: Twenty-one percent of patients had a bithermal caloric response, 62% had an absent ice water response, and 17% had an ice water response only 1 month following treatment. Mean follow-up was 27.3 months (range = 1-106 months). Eighty-four patients had one treatment course only. Fourteen patients required a further course of treatment due to recurrence of vertigo; 38% of these had recovery of caloric function. Significantly more patients with normal caloric function prior to initial treatment required further treatment compared to those with initial reduced caloric function (p < .05). Patients rendered absent ice water responsive were significantly less likely to require further treatment than those with a persistent caloric response (p < .0001). CONCLUSION: An absent ice water response is highly predictive of adequate vertigo control. Regimens of ITG that aim to completely ablate vestibular function are recommended. PMID- 10819106 TI - Eagle syndrome: classic and carotid artery types. AB - OBJECTIVE: To present a number of cases with elongated styloid process (Eagle) syndrome and to discuss the clinical presentation of this disorder. METHODS: A clinical study of patients operated on at King Abdul-Aziz University Hospital, Riyadh, from 1992 to 1996. RESULTS: The study group consisted of seven patients (six females and one male ranging in age from 20 to 43 years). Symptomatically, two distinct groups of patients could be identified: the classic syndrome (including three patients), which occurs in the tonsillectomized patients, and the styloid-carotid artery syndrome (including four patients), which occurs independently of tonsillectomy. In the first type, patients usually complain of spastic and nagging pain in the pharynx radiating to the mastoid region. In the second group, patients usually complain of attacks of syncope in association with pharyngeal pain referred to the course of the carotid artery. Diagnosis is made by history taking, palpating the tonsillar fossa, and radiographic demonstration of the process. Three-dimensional computed tomography reconstruction images were found to be very reliable in measuring the actual length of the styloid process and the stylohyoid ligament. CONCLUSION: This unusual disorder should be considered in the differential diagnosis of facial pain in some patients and as the cause of syncope in other patients. The paper discusses the embryologic, anatomic, pathogenetic, clinical, and therapeutic aspects of elongated styloid process with calcified stylohyoid ligament and the differential diagnosis is detailed. PMID- 10819107 TI - Management of benign thyroid nodules based on the findings of fine-needle aspiration. AB - Fine-needle aspiration (FNA) biopsy is a safe, simple, and inexpensive procedure that is particularly applicable for lesions of the head and neck. In our department, it is widely used for the evaluation of thyroid nodules. A total of 189 patients participated in a prospective study of benign thyroid nodules. Our aim was to verify the modality of treatment used in our department and the reliability of benign cytologic results of colloid goiter. All patients had cold nodules on thyroid scanning. The patients were divided into two groups, the first comprising 93 patients who did not undergo thyroid surgery and were followed up for 5 to 11 years. One case of malignancy was found in this group during the follow-up. The second group was comprised of 96 patients who were operated on despite FNA results of colloid goiter. Among them, five cases of malignancy were found. Four of these five cases could not be regarded as FNA failure. Our study confirms that the combination of clinical findings with those of the FNA is a reliable approach to the management of benign thyroid nodules. Nevertheless, long term follow-up is mandatory and repetitive aspirations should be considered. PMID- 10819108 TI - Squamous cell carcinoma of the nasal vestibule. AB - Squamous cell carcinoma of the nasal vestibule is a rare disease. Most advocate radiotherapy as a primary treatment for early tumours, with surgery reserved for salvage. For advanced disease, combined therapy with surgery and postoperative radiotherapy is generally recommended. Fourteen patients with squamous cell carcinoma of the nasal vestibule were reviewed. A classification of early versus late lesions was used. We achieved a 78% local regional control rate (minimum follow-up 3 years) in patients with early disease, with either radiotherapy or surgery as a primary modality of treatment. All patients with late disease recurred, requiring further surgical and/or radiation treatment. Only 20% of these patients were disease free at 2 years. Recurrent disease in either group, whether local or regional, carried a grave prognosis, with a 25% disease-free survival at 3 years. PMID- 10819109 TI - Basal cell adenocarcinoma: a rare malignancy of the salivary glands. PMID- 10819110 TI - Labyrinthine dysplasia, meningitis, and sensorineural deafness: a complex relationship. PMID- 10819111 TI - Sebaceous lymphadenoma of the parotid gland. PMID- 10819112 TI - Suction cautery use in lingual tonsillectomy. PMID- 10819113 TI - Peritonsillar abscess in a 2 1/2-month-old infant. PMID- 10819114 TI - Nasal natural killer/T cell lymphoma: case report with molecular biologic examination on Epstein-Barr virus and cell cycle regulatory p16, cyclin D1, Rb, and p53 genes. PMID- 10819115 TI - Congenital duplication in adult cervical esophagus. PMID- 10819116 TI - Implicit and explicit memory in autism: is autism an amnesic disorder? AB - Medial temporal lobe amnesic disorder is characterized by an impairment in explicit memory (e.g., remembering a shopping list) and intact implicit memory (e.g., a woman seems familiar although you cannot remember having met her before). This study examined whether children with high-functioning autism have this same dissociation between explicit and implicit memory abilities. Children with autism and normal development participated in three memory tasks: one implicit task (perceptual identification) and two explicit tasks (recognition and recall). Children with autism showed intact implicit and explicit memory abilities. However, they did not show the typical pattern of recalling more items from both the beginning and end of a list and instead only recalled items from the end of the list. These results do not support the theory that high functioning autism is a type of medial temporal lobe amnesia. However, these findings suggest that persons with autism use different organizational strategies during encoding or retrieval of items from memory. PMID- 10819117 TI - Recall for self and other in autism: children's memory for events experienced by themselves and their peers. AB - Research on memory processing suggests that memory for events that an individual experiences should be superior to that for similar events that someone else experiences (e.g., Baker-Ward et al., 1990). However, such predictions may not be applicable to individuals with autism. There are already suggestions that individuals with autism have specific difficulties in remembering (Boucher & Lewis, 1989). In addition, they are known to have more general difficulties involving processes related to the "self." If children with autism have difficulties in encoding information about themselves this could result in a deficit in personal episodic memory. The studies reported here compare memory for personally experienced events with that of memory for events experienced by a peer. An adaptation of a method devised by Boucher and Lewis has been employed to assess recall. Two separate studies were conducted to investigate whether children with autism are impaired at recalling personal events. Two groups of children took part in Study 1, a group of children with autism and a control group of typical children matched for verbal mental age. A group of children with moderate learning difficulties were employed in the second study to investigate whether the findings also occur in other groups of individuals who have learning disabilities. Findings indicate that, in the group with autism, events performed by the individual were recalled significantly less well than the observed events performed by a peer. However, the results for the nonautistic children in both studies showed that the opposite was true. Theoretical claims are discussed in the light of these findings. PMID- 10819118 TI - Schizophrenia with onset before the age of eleven: clinical characteristics of onset and course. AB - This study reports on the long-term course of 11 patients (6 girls, 5 boys) with childhood onset schizophrenia (COS, age at onset < 10 years). Patients were examined twice (mean follow-up period 38 years after onset). The premorbid development is assessed in terms of the Modified Premorbid Adjustment Scale (M PAS) and additionally described by distinct psychopathological categories. The psychopathology at the onset of psychosis and at the second follow-up examination was assessed by categorical application of the Positive and Negative Syndrome Scale (PANSS). The outcome was rated with the Disability Assessment Score (DAS). The course of psychotic episodes and intervals between them is presented according to DSM-IV subtype classifications. Ten of 11 patients presented premorbid developmental peculiarities that were not adequately covered by the M PAS subscales. Whereas in the 4 patients with acute onset of psychosis the positive PANSS-type was predominant, in the 7 patients with an insidious onset the negative PANSS-type prevailed. The nature of the diagnostic subtypes varied markedly across the course of the illness. In case of a continuous predominant catatonic symptomatology the outcome was poor. Detailed case descriptions help to illuminate the heterogeneous psychopathology of COS. Various temporary premorbid behavioral peculiarities were precursors of COS. A differentiation between premorbid and prodromal signs proved to be arbitrary. Our results contradict the assumption that COS is characterized only by a negative symptomatology. PMID- 10819119 TI - Review of adaptive behavior studies in mentally retarded persons with autism/pervasive developmental disorder. AB - A review is presented of the investigations carried out concerning the adaptive behavior of persons with the dual disability of mental retardation and autism/PDD. A close correspondence is found between the results obtained by means of a Dutch set of scales, the SRZ, SGZ, and SMZ, and those obtained by means of the Vineland Adaptive Behavior Scales. Compared with matched nonautistic persons, the performance of dually disabled persons is found to be particularly poor in the domain Social Skills/Socialization and somewhat less poor in the domain Communication. Performance of the two categories does not differ in the domains Self Help/Daily Living Skills and Gross Motor Skills/Motor Skills. As a net result, persons of the dually disabled category are found to obtain comparatively low total scores for social competency, the SRZ-Total/Adaptive Behavior Composite. In addition, autistic mentally retarded persons are found to display more maladaptive behavior. For comparative studies in the mental retardation field the use of level of Self Help/Daily Living Skills and, to a lesser extent, level of Gross Motor Skills/Motor Skills are recommended as suitable matching variables. PMID- 10819121 TI - Brief report: A longitudinal study of quality of life and independence among adult men with autism. PMID- 10819120 TI - Early development, temperament, and functional impairment in autism and fragile X syndrome. AB - We compared the developmental status, functional abilities, and temperament of 31 young boys with fragile X syndrome (FXS) who did not have autism, matched on chronological age, gender, and race, with 31 boys with autism but no FXS. Children with autism exhibited a more variable profile of development in comparison with a relatively flat profile for children with FXS. Children with autism were significantly more delayed in social skills and were rated by observers as exhibiting a greater degree of impairment in cognitive, communication, and social skills. On temperament ratings, both groups were slower to adapt, less persistent, and more withdrawing than the reference group. Boys with FXS were rated as more active than the referent group, whereas boys with autism were rated as less intense, more distractible, having a higher threshold for response, and less rhythmic than the reference group. A smaller three-group analysis compared boys with FXS, boys with autism, and boys with both FXS and autism. Children with both autism and FXS were substantially more delayed than children with autism or FXS alone. PMID- 10819122 TI - Brief report: Brief syntactic analysis in Asperger syndrome: a preliminary study. PMID- 10819123 TI - Regarding secretin for treating autistic disorder. PMID- 10819124 TI - Secretin in autism--a parent's perspective. PMID- 10819125 TI - Intravenous immunoglobulin treatment in autism. PMID- 10819126 TI - PDDNOS in DSM-IV. PMID- 10819127 TI - Progress with PETS: continuing concerns and experience to date. PMID- 10819129 TI - Looking to the future: educational issues debated at council seminar 2000. PMID- 10819128 TI - How frequently should dogs and cats be vaccinated? PMID- 10819130 TI - Descriptive epidemiology of scrapie in Great Britain: results of a postal survey. AB - In 1998, a questionnaire was sent to 11,554 British sheep farmers to determine how many believed that scrapie cases had occurred in their flock; 61.4 per cent of them responded anonymously. The results indicated that 14.9 per cent of farmers with more than 30 breeding ewes thought that they had ever experienced scrapie in their flock and 2.7 per cent thought that they had had cases in the past 12 months. A comparison of these results with the number of farmers reporting suspect scrapie cases to MAFF, in accordance with the statutory requirement, suggests that only 13 per cent of farmers who suspect that they may have cases of scrapie are currently reporting them. Scrapie occurred in all regions of the country but there was an apparent regional variation. Larger farms and those with purebred sheep appeared to be at greater risk of having cases. Other differences between affected and unaffected farms included lambing practices and sheep purchasing policy. On the majority of farms the first case occurred in a purchased animal. The survey also revealed a need for the provision of further information about scrapie to farmers. PMID- 10819131 TI - Effects of iron dextran injections on the incidence of abomasal bloat, clinical pathology and growth rates in lambs. AB - A preliminary study revealed significantly lower serum iron concentrations in lambs that developed abomasal bloat about one week later, than in lambs that did not develop bloat. In a subsequent trial, with 754 naturally reared twin lambs from five flocks, iron dextran injections were found to have a preventive effect on the development of abomasal bloat. Clinical signs of abomasal bloat were observed in the placebo-treated lamb of 16 couples and in the iron-treated lamb of six couples (P<0.05). In three further couples, both lambs developed bloat. The iron-treated group had significantly (P<0.001) better average weight gains, both from birth to summer, and from birth to autumn of approximately 0.5 and 1 kg, respectively. There was a reduction in red blood cell values and iron saturation in the placebo group 14 days after treatment but not in the iron treated lambs. In one of the flocks, there was a decrease in the cumulative incidence of abomasal bloat from 37 per cent to 3 per cent during the period of four years after measures such as later lambing, earlier turnout and iron injections were introduced. PMID- 10819132 TI - Clinical and laboratory observations in 91 dogs infected with Dirofilaria immitis in northern Greece. AB - The medical records of 91 dogs with heartworm (Dirofilaria immitis) infection were reviewed, and diagnoses were established by using parasitological and immunological methods. Twenty-one animals were asymptomatic (stage I), 57 had mild to moderate clinical signs (stage II), and 13 had the severe form of the disease including right congestive heart failure and the caval syndrome (stage III). Thoracic radiography revealed right ventricular enlargement in 38 of the dogs, pulmonary vascular enlargement in 43, and parenchymal lesions in 27. Only the cardiac and vascular changes were correlated positively with the clinical stages. D. immitis microfilaraemia was detected in 75 of 85 dogs. Occult infection occurred only in eight stage II and two stage III dogs. Thirty-two of the dogs were treated with thiacetarsamide and 39 were treated with melarsomine, and no differences were found in terms of drug efficacy or complication rate; nine stage II dogs suffered pulmonary thromboembolism and one suffered acute liver disease and there were six fatalities. The 50 treated dogs in stages II and III which were followed up for six months all recovered completely. The performance of 38 of 61 working dogs was completely restored, and the performance of another four was partially restored. PMID- 10819133 TI - Development of vitrified matured cattle oocytes after thawing and culture in vitro. AB - Bovine oocytes were partly denuded either at the beginning (t0) or six hours (t6) after the beginning of maturation and vitrified by the open pulled straw method at the end of the maturation process. After warming and fertilisation, their development in vitro and in vivo was assessed. The rates of production of blastocysts achieved in vitro were 3.4 per cent for the t0 group and 0.9 per cent for the t6 group compared with 40.4 per cent for the control oocytes. After transfer at the blastocyst stage pregnancies have been established in the three groups. Some of these pregnancies originated from vitrified oocytes which were further vitrified at the blastocyst stage before being transferred into synchronised recipients. PMID- 10819134 TI - Ileal muscular hypertrophy and rupture in a pony three years after surgery for ileocaecal intussusception. PMID- 10819135 TI - Serological evidence of the absence of peste des petits ruminants in Tanzania. PMID- 10819136 TI - Veterinary surveillance. PMID- 10819137 TI - Pet travel scheme. PMID- 10819138 TI - Exotic pets. PMID- 10819139 TI - Tendering for LVI work. PMID- 10819140 TI - Sebaceous adenitis in standard poodles. PMID- 10819141 TI - Thermal tolerance, climatic variability and latitude. AB - The greater latitudinal extents of occurrence of species towards higher latitudes has been attributed to the broadening of physiological tolerances with latitude as a result of increases in climatic variation. While there is some support for such patterns in climate, the physiological tolerances of species across large latitudinal gradients have seldom been assessed. Here we report findings for insects based on published upper and lower lethal temperature data. The upper thermal limits show little geographical variation. In contrast, the lower bounds of supercooling points and lower lethal temperatures do indeed decline with latitude. However, this is not the case for the upper bounds, leading to an increase in the variation in lower lethal limits with latitude. These results provide some support for the physiological tolerance assumption associated with Rapoport's rule, but highlight the need for coupled data on species tolerances and range size. PMID- 10819142 TI - Environmental colour affects aspects of single-species population dynamics. AB - Single-species populations of ciliates (Colpidium and Paramecium) experienced constant temperature or white or reddened temperature fluctuations in aquatic microcosms in order to test three hypotheses about how environmental colour influences population dynamics. (i) Models predict that the colour of population dynamics is tinged by the colour of the environmental variability. However, environmental colour had no effect on the colour of population dynamics. All population dynamics in this experiment were reddened, regardless of environmental colour. (ii) Models predict that populations will track reddened environmental variability more closely than white environmental variability and that populations with a higher intrinsic growth rate (r) will track environmental variability more closely than populations with a low r. The experimental populations behaved as predicted. (iii) Models predict that population variability is determined by interaction between r and the environmental variability. The experimental populations behaved as predicted. These results show that (i) reddened population dynamics may need no special explanation, such as reddened environments, spatial subdivision or interspecific interactions, and (ii) and (iii) that population dynamics are sensitive to environmental colour, in agreement with population models. Correct specification of the colour of the environmental variability in models is required for accurate predictions. Further work is needed to study the effects of environmental colour on communities and ecosystems. PMID- 10819143 TI - Signals and behavioural responses are not coupled in males: aggression affected by replacement of an evolutionarily lost colour signal. AB - Male Sceloporus virgatus lack the blue abdominal patches which are used during aggressive encounters in other Sceloporus lizards. Herein we report that, despite having lost this signal, males have retained a behavioural response to experimentally restored blue abdominal patches. We tested two adaptive hypotheses: selection acted primarily upon signallers or selection acted upon both signallers and receivers. The first predicts that only the signal is lost and that male interactions should be affected by the restoration of blue patches. The latter predicts that both the signal and behavioural response are lost and the display of the restored blue patches should have no effect on male-male interactions. We compared the behaviour of receivers in paired encounters where one male (signaller) had blue-painted abdominal patches to a set of trials where both males had white-painted abdomens, unmanipulated abdomens or a novel-painted pattern. The receivers of the blue-painted signal were more likely to display submissive behaviour. The receivers in either the unmanipulated, white-painted or novel-painted signal trials were more likely to display neutral behaviour. These results support the hypothesis that receivers have retained a behavioural response and selection has acted primarily on the signaller. We believe this is the first documentation of males responding to an evolutionarily lost signal in conspecific males. PMID- 10819144 TI - Energetics, cost reduction and functional consequences of fish morphology. AB - Cost reduction strategies are often invoked as explanations when studies of adaptation fail to find predicted costs. This might seem discouraging, offering little opportunity for further investigation. In this paper, we demonstrate that cost reduction strategies can themselves be investigated by arguments from design. Recent work on inducible morphological defences has shown that hydrodynamical disadvantages (e.g. high drag) in fishes can be compensated for by standard metabolic rate (SMR) adjustments. Here, we theoretically investigate the possibilities and limitations for swimming cost compensation through SMR adjustment. We continue by modelling how intraspecific power curve variation affects the optimal swimming velocity between food patches. Our results show that, even though SMR modifications may compensate for hydrodynamical disadvantages, low-drag fishes will nevertheless have a marked advantage under high food abundance. The relative advantage will decrease with decreasing food levels. We also show that hydrodynamical properties of fishes can be used to predict their propensity to become foraging (or swimming) specialists. Low-drag fishes can use a broad range of swimming velocities without substantial increases in swimming cost, whereas the cost of deviating from the optimal swimming velocity increases markedly in high-drag fishes. The results have important implications for the evolution of morphological diversity in fishes. PMID- 10819145 TI - Ants estimate area using Buffon's needle. AB - We show for the first time, to our knowledge, that ants can measure the size of potential nest sites. Nest size assessment is by individual scouts. Such scouts always make more than one visit to a potential nest before initiating an emigration of their nest mates and they deploy individual-specific trails within the potential new nest on their first visit. We test three alternative hypotheses for the way in which scouts might measure nests. Experiments indicated that individual scouts use the intersection frequency between their own paths to assess nest areas. These results are consistent with ants using a 'Buffon's needle algorithm' to assess nest areas. PMID- 10819146 TI - Polymorphic growth rates in myrmecophilous insects. AB - A polymorphism in growth rates was recently described affecting the larval development of the myrmecophilous butterfly Maculinea rebeli, spanning different years in a single insect population. The close integration of M. rebeli into the host ant colonies, facilitated by adaptations in behaviour and chemical mimicry, make extended larval development a successful strategy. Here we present additional data for M. rebeli and new data for Maculinea alcon (another cuckoo feeding lycaenid) and the two myrmecophilous predators Maculinea arion and Microdon mutabilis (Diptera: Syrphidae). As predicted, M. alcon shows the same growth pattern as M. rebeli with a proportion of caterpillars developing in one year and the remainder over two years. This pattern holds in both northern and southern European populations, where M. alcon exploits different species of host. Against expectation, the same bimodal distribution of pre-pupation body weights, indicating one and two year developers, was found for the larvae of M. arion and M. mutabilis. As predators, both species are less closely integrated in their host ant colonies, suggesting that the polymorphism in growth rates is a more general adaptation to a myrmecophilous life style, arrived at by convergent evolution between the Maculinea and Microdon species. For predatory species we suggest that biennialism is an adaptation to the migratory behaviour of the host made possible by the predators' ability to fast over extended periods. We also hypothesize that M. arion represents an ancestral strategy in Maculinea butterflies and that the growth polymorphism might have become genetically fixed in the cuckoo-feeding species. PMID- 10819147 TI - Invading parasites cause a structural shift in red fox dynamics. AB - The influence of parasites on host life histories and populations is pronounced. Among several diseases affecting animal populations throughout the world, sarcoptic mange has influenced many carnivore populations dramatically and during the latest epizootic in Fennoscandia reduced the abundance of red fox by over 70%. While the numerical responses of red fox populations, their prey and their competitors as well as clinical implications are well known, knowledge of how sarcoptic mange affects the structure of the dynamics of red fox populations is lacking. Integrating ecological theory and statistical modelling, we analysed the long-term dynamics (1955-1996) of 14 Danish red fox populations. As suggested by the model, invading sarcoptic mange significantly affected direct and delayed density dependence in red fox dynamics and concomitant shifts in fluctuation patterns were observed. Our statistical analyses also revealed that the spatial progressive spread of mange mites was mirrored in the autocovariate structures of red fox populations progressively exposed to sarcoptic mange. PMID- 10819148 TI - Manipulation of host behaviour by parasites: a weakening paradigm? AB - New scientific paradigms often generate an early wave of enthusiasm among researchers and a barrage of studies seeking to validate or refute the newly proposed idea. All else being equal, the strength and direction of the empirical evidence being published should not change over time, allowing one to assess the generality of the paradigm based on the gradual accumulation of evidence. Here, I examine the relationship between the magnitude of published quantitative estimates of parasite-induced changes in host behaviour and year of publication from the time the adaptive host manipulation hypothesis was first proposed. Two independent data sets were used, both originally gathered for other purposes. First, across 137 comparisons between the behaviour of infected and uninfected hosts, the estimated relative influence of parasites correlated negatively with year of publication. This effect was contingent upon the transmission mode of the parasites studied. The negative relationship was very strong among studies of parasites which benefit from host manipulation (transmission to the next host occurs by predation on an infected intermediate host), i.e. among studies which were explicit tests of the adaptive manipulation hypothesis. There was no correlation with year of publication among studies on other types of parasites which do not seem to receive benefits from host manipulation. Second, among 14 estimates of the relative, parasite-mediated increase in transmission rate (i.e. increases in predation rates by definitive hosts on intermediate hosts), the estimated influence of parasites again correlated negatively with year of publication. These results have several possible explanations, but tend to suggest biases with regard to what results are published through time as accepted paradigms changed. PMID- 10819149 TI - Life history of a malaria parasite (Plasmodium mexicanum): independent traits and basis for variation. AB - Plasmodium mexicanum, a malaria parasite of lizards, exhibits substantial variation among infections in the life-history traits which define its blood dwelling stages. Such variation in life histories among infections is common in Plasmodium and may influence the ecology and evolution of the parasite's transmission success and virulence. Insight into these issues requires identification of independent traits (some traits may be bound by developmental trade-offs) and the importance of genetic versus host effects producing the variation. We studied 11 life-history traits in 120 induced infections of P. mexicanum in its natural lizard host (20 each from six donor infections). The traits varied among infections and fell into three clusters: rate/peak (rate of increase and peak parasitaemia of asexuals and gametocytes), time (duration of pre-patent period and the infection's growth) and maturity (timing of first gametocytes). Thus, few life-history traits define an infection in the lizard's blood. Donor effects were significant for ten traits and two trait clusters (maturity was the exception) suggesting genetic differences among infections may influence the rate of increase and peak parasitaemia, but not the timing of the first production of gametocytes. PMID- 10819150 TI - Inbreeding avoidance and reproductive skew in a cooperative mammal. AB - In animal social groups, socially subordinate individuals frequently show low reproductive success or completely fail to breed. This suppression of subordinate reproduction is currently typically attributed to control by dominant individuals. However, subordinates in cooperative groups often lack access to unrelated mates, and an alternative possibility is that their reproduction is limited by inbreeding avoidance. Using the eusocial Damaraland mole-rat Cryptomys damarensis, this paper provides the first experimental evidence, to our knowledge, for this explanation. Subordinate, non-breeding female mole-rats were given access to unrelated mates while remaining in the presence of dominant females, and many became reproductively active soon after unrelated males were introduced. Inbreeding avoidance and the availability of unrelated mates provides a plausible and untested explanation for variation in reproductive skew across animal societies. PMID- 10819151 TI - Nuptial gifts and the use of body resources for reproduction in the green-veined white butterfly Pieris napi. AB - Reproduction in butterflies, as in many holometabolous insects, is usually constrained by the amount of nutrients the animals can collect as juveniles. In polyandric species the females can also supplement their larval-derived reserves with protein-rich donations, so-called nuptial gifts, delivered by the males at mating. Recent findings also indicate that females have access to nitrogen from the histolysis of flight muscles in the thorax. This field study examined how butterflies of the polyandric gift-giving species Pieris napi (Lepidoptera: Pieridae) use body resources in their reproduction and how the male donations affect the females use of stored reserves. The results support earlier studies, indicating that females use resources from the breakdown of thorax muscles to increase their reproductive potential and the results also indicate that males also use thorax material in their reproduction. The study also supports recent findings that the male donation increases the breakdown of body resources and thereby boosts the reproductive output of the female. PMID- 10819152 TI - Divorce in cooperatively breeding long-tailed tits: a consequence of inbreeding avoidance? AB - The decision of whether to divorce a breeding partner between reproductive attempts can significantly affect individual fitness. In this paper, we report that 63% of surviving pairs of long-tailed tits Aegithalos caudatus divorced between years. We examine three likely explanations for the high divorce rate in this cooperative breeder. The 'better option' hypothesis predicts that divorce and re-pairing increases an individual's reproductive success. However, divorcees did not secure better partners or more helpers and there was no improvement in their reproductive success following divorce. The 'inbreeding avoidance' hypothesis predicts that females should disperse from their family group to avoid breeding with philopatric sons. The observed pattern of divorce was consistent with this hypothesis because, in contrast to the usual avian pattern, divorce was typical for successful pairs (81%) and less frequent in unsuccessful pairs (36 43%). The 'forced divorce' hypothesis predicts that divorce increases as the number of competitors increases. The pattern of divorce among failed breeders was consistent with this hypothesis, but it fails to explain the overall occurrence of divorce because divorcees rarely re-paired with their partners' closest competitors. We discuss long-tailed tits' unique association between divorce and reproductive success in the context of dispersal strategies for inbreeding avoidance. PMID- 10819153 TI - A skew model for the evolution of sociality via manipulation: why it is better to be feared than loved. AB - Concession-based reproductive skew models predict that social groups can form via persuasion, whereby dominant individuals forfeit some reproduction to subordinates as an incentive to stay and help. We have developed an alternative skew model based on manipulation, whereby dominant individuals coerce subordinates into staying and helping by imposing costs on their independent reproductive prospects. Stable groups can evolve under a much wider range of genetic and ecological conditions under this manipulation model than under concession models. We describe evidence that various forms of pre-emptive and ongoing manipulation occur in nature and we discuss the implications of the model for the development of a general theory of social evolution. PMID- 10819154 TI - Spontaneous number representation in semi-free-ranging rhesus monkeys. AB - Previous research has shown that animals possess considerable numerical abilities. However, this work was based on experiments involving extensive training, a small number of captive subjects and relatively artificial testing procedures. We present the results of experiments on over 200 semi-free-ranging rhesus monkeys using a task which involves no training and mimics a natural foraging problem. The subjects observed two experimenters place pieces of apple, one at a time, into each of two opaque containers. The experimenters then walked away so that the subjects could approach. The monkeys chose the container with the greater number of apple slices when the comparisons were one versus two, two versus three, three versus four and three versus five slices. They failed at four versus five, four versus six, four versus eight and three versus eight slices. Controls established that it was the representation of number which underlay their successful choices rather than the amount of time spent placing apple pieces into the box or the volume of apple placed in the box. The failures at values greater than three slices stand in striking contrast to other animal studies where training was involved and in which far superior numerical abilities were demonstrated. The range of success achieved by rhesus monkeys in this spontaneous-number task matches the range achieved by human infants and corresponds to the range encoded in the syntax of natural languages. PMID- 10819155 TI - Adherence and drug resistance: predictions for therapy outcome. AB - We combine standard pharmacokinetics with an established model of viral replication to predict the outcome of therapy as a function of adherence to the drug regimen. We consider two types of treatment failure: failure to eliminate the wild-type virus, and the emergence of drug-resistant virus. Specifically, we determine the conditions under which resistance dominates as a result of imperfect adherence. We derive this result for both single- and triple-drug therapies, with attention to conditions which favour the emergence of viral strains that are resistant to one or more drugs in a cocktail. Our analysis provides quantitative estimates of the degree of adherence necessary to prevent resistance. We derive results specific to the treatment of human immunodeficiency virus infection, but emphasize that our method is applicable to a range of viral or other infections treated by chemotherapy. PMID- 10819156 TI - [4-[[N-(3-chlorophenyl)carbamoyl]oxy]-2-butynyl]-trimethylammonium (McN-A-343) related compounds. Effect of the butynyl chain inclusion into an aromatic unit on the potency for muscarinic receptors. AB - A series of derivatives of the known M1 selective muscarinic receptor agonist McN A-343 (1) was designed with the aim of investigating the effects of structural variations on both the butynyl chain and the phenyl ring of 1. The butynyl chain was replaced with an aromatic spacer, and the effects of such a modification on the stereoelectronic properties of the molecules were theoretically studied and considered compatible with muscarinic receptor affinity. Substituents on the phenyl ring of 1 were selected so as to vary their electronic and hydrophobic properties. This design strategy did not produce muscarinic M1 receptor agonists more potent than the prototype 1, even if some analogues displayed functional selectivity for different muscarinic receptor subtypes. Compounds 3 and 7 were selective agonists towards muscarinic M3 receptors, while compounds 14, 16 and 18 were selective muscarinic M2 receptor agonists. The most interesting derivative was 8, a full agonist at muscarinic M3 receptors devoid of activity at both muscarinic M1 and M2 subtypes. The pharmacological profile of the series was further characterized by studying the anticholinesterase and miotic activities of some representative compounds. Compounds 3-8 turned out to be weak acetylcholinesterase inhibitors, while derivatives 4, 6, 8 and 11 were able to significantly reduce the pupillary diameter in rabbit, indicating 8 as an effective miotic agent. PMID- 10819157 TI - Inhibitors of the fungal cell wall. Synthesis of 4-aryl-4-N-arylamine-1-butenes and related compounds with inhibitory activities on beta(1-3) glucan and chitin synthases. AB - As part of our project devoted to the search for antifungal agents, which act via a selective mode of action, we synthesized a series of new 4-aryl- or 4-alkyl-N arylamine-1-butenes and transformed some of them into 2-substituted 4-methyl tetrahydroquinolines and quinolines by using a novel three-step synthesis. Results obtained in agar dilution assays have shown that 4-aryl homoallylamines not possessing halogen in their structures, tetrahydroquinolines and quinolines, display a range of antifungal properties in particular against Epidermophyton floccosum and Microsporum canis. Regarding the mode of action, all active compounds showed in vitro inhibitory activities against beta(1-3) glucan-synthase and mainly against chitin-synthase. These enzymes catalyze the synthesis of beta(1-3) glucan and chitin, respectively, major polymers of the fungal cell wall. Since fungal but not mammalian cells are encased in a cell wall, its inhibition may represent a useful mode of action for these antifungal compounds. PMID- 10819158 TI - 3Beta-hydroxy-6-aza-cholestane and related analogues as phosphatidylinositol specific phospholipase C (PI-PLC) inhibitors with antitumor activity. AB - 6-Aza steroid analogues were synthesized as PI-PLC inhibitors. The most active compound, 3beta-hydroxy-6-aza-cholestane (1) showed potent PI-PLC inhibition (IC50 = 1.8 microM), similar to that of the commercially available steroid analogue U73122 (IC50 = 1-2.1 microM). Compound 1 exhibited significant growth inhibition effects (IC50 = 1.3 microM in each case) against MCF-7 and HT-29 cancer cells in in vitro cell culture. Compound 1 also inhibited the in vitro adhesion and transmigration of HT-1080 fibrosarcoma cells at 2.5 and 5.0 microM, respectively. In vivo, compound 1, at 1 mg/kg/day, reduced the volume of MCF-7 tumors in xenograft models, without weight loss in mice. Structure activity relationships of this series of compounds revealed that a hydrophobic cholesteryl side chain, 3beta-hydroxy group and a C-6 nitrogen containing a hydrogen atom at position-6 are crucial for activity. N-Maleic amidoacid derivative 11 also exhibited weak inhibition (IC50 = 16.2 microM). PMID- 10819159 TI - Acyloxymethyl as a drug protecting group. Part 6: N-acyloxymethyl- and N [(aminocarbonyloxy)methyl]sulfonamides as prodrugs of agents containing a secondary sulfonamide group. AB - Tertiary N-acyloxymethyl- and N-[(aminocarbonyloxy)methyl]sulfonamides were synthesised and evaluated as novel classes of potential prodrugs of agents containing a secondary sulfonamide group. The chemical and plasma hydrolyses of the title compounds were studied by HPLC. Tertiary N-acyloxymethylsulfonamides are slowly and quantitatively hydrolysed to the parent sulfonamide in pH 7.4 phosphate buffer, with half-lives ranging from 20 h, for 7d, to 30 days, for 7g. Quantitative formation of the parent sulfonamide also occurs in human plasma, the half-lives being within 0.2-2.0 min for some substrates. The rapid rate of hydrolysis can be ascribed to plasma cholinesterase, as indicated by the complete inhibition observed at [eserine] = 0.10 mM. These results suggest that tertiary N acyloxymethylsulfonamides are potentially useful prodrugs for agents containing a secondary sulfonamide group, especially with pKa < 8, combining a high stability in aqueous media with a high rate of plasma activation. In contrast, N [(aminocarbonyloxy)methyl]sulfonamides 7h-j do not liberate the parent sulfonamide either in aqueous buffers or in human plasma and thus appear to be unsuitable for development as sulfonamide prodrugs. PMID- 10819160 TI - Chemical and enzymatic synthesis of fructose analogues as probes for import studies by the hexose transporter in parasites. AB - Various D-fructose analogues modified at C-1 or C-6 positions were synthesized from D-glucose by taking advantage of the Amadori rearrangement or using the aldol condensation between dihydroxyacetone phosphate and appropriate aldehyde catalyzed by fructose 1,6-diphosphate aldolase from rabbit muscle. The affinities of the analogues for the glucose transporter expressed in the mammalian form of Trypanosoma brucei were determined by inhibition of radiolabelled 2-deoxy-D glucose (2-DOG) transport using zero-trans kinetic analysis. Interestingly, the analogues bearing an aromatic group (i.e. a fluorescence marker) at C-1 or C-6 positions present comparable apparent affinities to D-fructose for the transporter. This result could find applications for hexose transport studies and also provides criteria for the design of glucose import inhibitors. PMID- 10819161 TI - Characterization of fertilin beta-disintegrin binding specificity in sperm-egg adhesion. AB - An essential step leading to fertilization is the binding of a sperm to the egg plasma membrane. Fertilin beta, a membrane bound protein on the extracellular surface of sperm, partially mediates this binding via the alpha6beta1 integrin. Fertilin beta is a member of the still expanding family of ADAM proteins (a disintegrin and metalloprotease) that are implicated in many cellular functions ranging from neurogenesis to myoblast fusion and cytokine processing. Fertilin beta contains a highly conserved motif (D/E)ECD in the disintegrin domain. This suggests that (D/E)ECD could be the consensus sequence for recognition of disintegrins by alpha6beta1 integrins. Previously, it has been demonstrated that small peptides containing different moieties of this consensus sequence are inhibitors of in vitro fertilization. In the present study, we sought to determine whether a four amino acid peptide sequence with two adjacent acidic residues improved inhibition, and investigated the importance for inhibition of a cysteine versus a cystine. A series of linear and cyclic peptides were synthesized, in which either one or both adjacent acidic residues in the sequence DECD were mutated to their corresponding amides (N or Q). To explore the required oxidation state of the cysteine in the (D/E)ECD sequence, it was protected as a mixed disulfide. Our results indicate that only one acidic residue is required for inhibition of fertilization and a reduced C is required. PMID- 10819162 TI - Lipase-catalysed synthesis of new acetylcholinesterase inhibitors: N benzylpiperidine aminoacid derivatives. AB - New acetylcholinesterase inhibitors were synthetized via a lipase-mediated regioselective amidation using Candida antarctica lipase B as a biocatalyst in the key step. The new compounds have two different structural fragments: a N benzylpiperidine moiety to anchor the enzyme active site and a dicarboxylic aminoacid to act as a biological carrier. Some analogues of N-benzylpiperazine were also synthesised and studied but they did not display AChE inhibitor activity. A preliminary structure activity relationship study was performed employing some computational techniques as similarity indices and electrostatic potential maps. PMID- 10819163 TI - Structural studies on bioactive compounds. Part 29: palladium catalysed arylations and alkynylations of sterically hindered immunomodulatory 2-amino-5 halo-4,6-(disubstituted)pyrimidines. AB - The immunological agent bropirimine 5 is a tetra-substituted pyrimidine with anticancer and interferon-inducing properties. Synthetic routes to novel 5-aryl analogues of bropirimine have been developed and their potential molecular recognition properties analysed by molecular modelling methods. Sterically challenged 2-amino-5-halo-6-phenylpyrimidin-4-ones (halo = Br or I) are poor substrates for palladium catalysed Suzuki cross-coupling reactions with benzeneboronic acid because the basic conditions of the reaction converts the amphoteric pyrimidinones to their unreactive enolic forms. Palladium-mediated reductive dehalogenation of the pyrimidinone substrates effectively competes with cross-coupling. 2-Amino-5-halo-4-methoxy-6-phenylpyrimidines can be converted to a range of 5-aryl derivatives with the 5-iodopyrimidines being the most efficient substrates. Hydrolysis of the 2-amino-5-aryl-4-methoxy-6-phenylpyrimidines affords the required pyrimidin-4-ones in high yields. Semi-empirical quantum mechanical calculations show how the nature of the 5-substituent influences the equilibrium between the 1H- and 3H-tautomeric forms, and the rotational freedom about the bond connecting the 6-phenyl group and the pyrimidine ring. Both of these factors may influence the biological properties of these compounds. PMID- 10819164 TI - Synthesis, antiplatelet activity and comparative molecular field analysis of substituted 2-amino-4H-pyrido[1,2-a]pyrimidin-4-ones, their congeners and isosteric analogues. AB - 2-(1-Piperazinyl)-4H-pyrido[1,2-a]pyrimidin-4-one (5a) is a recently described in vitro inhibitor of human platelet aggregation which specifically inhibits the activity of high affinity cAMP phosphodiesterase. A number of substitution derivatives, isosteres, and analogues of 5a were now synthesized and tested in vitro for their inhibitory activity on human platelet aggregation induced in platelet-rich plasma by ADP, collagen, or the Ca2+ ionophore A23187. Among the most effective compounds, the 6-methyl, 8-methyl and 6,8-dimethyl derivatives of 5a resulted nearly as active as the lead when platelet aggregation was induced by ADP or A23187, but less active when collagen was the inducer. On the basis of present results and those previously obtained by us in this and 2-aminochromone structural fields, we have developed a statistically significant 3-D QSAR model, using comparative molecular field analysis (CoMFA), describing the variation of the antiplatelet activity in terms of molecular steric and electrostatic potential changes. PMID- 10819165 TI - Dialkylaminoalkylnaphthalenes as novel opioid-like analgesics. AB - The study of dialkylaminoalkylnaphthalenes as novel opioid-like analgesics is reported. In particular, the synthesis of (1R,2R/1S,2S)-1-ethyl-1-[2-(6 hydroxynaphthyl)]-1-hydrox-2-m ethyl-2-dimethylaminoethane and its structural analogue (1R,2R/1S,2S)-1-ethyl-1-[2-(6-fluoronaphthyl)]-1-hydroxy-2-methyl- 2 dimethylaminoethane and the configurational analysis by X-ray and 1H NMR spectroscopy are described. Pharmacological profiles are discussed on the basis of the experimental results of analgesia tests (hot plate and writhing test) and rota-rod test, which was performed to distinguish analgesia from drug-induced motor changes. The compounds showed dose-dependent antinociception, with less potency than morphine. Motor coordination appeared to be less involved. PMID- 10819166 TI - Recognition and cleavage of DNA by rebeccamycin- or benzopyridoquinoxaline conjugated of triple helix-forming oligonucleotides. AB - Indolocarbazole and benzopyridoquinoxaline derivatives have been shown to have anti-tumor activity and to stimulate DNA topoisomerase I-mediated cleavage. Two indolocarbazole compounds (R-6 and R-95) and one benzopyridoquinoxaline derivative (BPQ(1256)) were covalently attached to the 3'-end of a 16mer triple helix-forming oligonucleotide (TFO). These conjugates bind to DNA with a higher affinity than the unsubstituted oligonucleotides. Furthermore, they induce topoisomerase I-mediated and triplex-directed DNA cleavage in a sequence-specific manner. PMID- 10819167 TI - Triarylethylene bisphenols with a novel cycle are ligands for the estrogen receptor. AB - We have prepared a series of triarylethylene and triarylethane systems, analogues of the selective antiestrogen tamoxifen, in which the alkyl substituent is tethered to the distal, rather than the proximal aryl ring by a 5-, 6-, or 7 membered carbocycle. This unusual cyclic structure rigidifies the ligand and adds bulk in a manner that is different from the more typical cyclization to the proximal aryl ring, as in the antiestrogen nafoxidine. These new systems were prepared efficiently by the addition of a benzylic sodium reagent, generated from the corresponding chloride by treatment with sodium naphthalenide, to a doubly protected 4,4'-dihydroxybenzophenone, followed by dehydration and deprotection. In all cases, formation of the exocyclic alkene was preferred. Two of the corresponding alkanes could be obtained by catalytic hydrogenation. All of these compounds have relatively high binding affinity for the estrogen receptor, and some of them demonstrate a significant level of affinity selectivity for the estrogen receptor alpha subtype. Accommodation of these newly rigidified cyclic triarylethylene systems into the ligand-binding pocket of the estrogen receptor can be visualized by molecular modeling. PMID- 10819168 TI - The use of 'electronic nose' sensor responses to predict the inhibition activity of alcohols on the cytochrome P-450 catalyzed p-hydroxylation of aniline. AB - A quantitative structure activity relationship (QSAR) has been formulated to describe the inhibitory action of a series of alcohols on the cytochrome P-450 catalyzed p-hydroxylation of aniline. The descriptors used in the QSAR are the responses of individual sensors in a polymer-based electronic nose, and are all easily generated experimental values. If the various electronic nose sensor response patterns for the family of test alcohols reflect differences in the chemical properties that are involved in the cytochrome P-450 inhibition process, it ought to be possible to correlate the differences in the electronic nose signals of these analytes with the differences in the cytochrome P-450 inhibition by these species. To evaluate this possibility, multiple linear regression was performed on data obtained from exposure of a series of test alcohols to 19 sensors of a conducting polymer composite electronic nose array. A genetic algorithm was then used to select the optimal set of sensors that best described the inhibitory activity of these alcohols within a linear regression model. The regression equation fit the inhibition data of 20 of the alcohols with an R of 0.995. This fit compares favorably with previously published QSARs on this system that have used log P (P triple bond octanol-water partition coefficient) along with steric parameters of the alcohols, and also compares favorably to QSARs formulated using theoretically calculated parameters. PMID- 10819169 TI - Inhibition of protein phosphatases by Michael adducts of ascorbic acid analogues with alpha,beta-unsaturated carbonyl compounds. AB - By investigating the stereospecific Michael reaction of derivatives of ascorbic acid with acrolein we obtained a novel class of protein Ser/Thr phosphatase inhibitors. The inhibitory effect of the Michael adducts was examined using the canonical protein phosphatases type 1, 2A and 2B. Of the isozymes examined the type 1 isoform was strongly inhibited. PMID- 10819170 TI - Hippuryl-alpha-methylphenylalanine and hippuryl-alpha-methylphenyllactic acid as substrates for carboxypeptidase A. Syntheses, kinetic evaluation and mechanistic implication. AB - (R)- and (S)-Hippuryl-alpha-methylphenylalanine [(R)- and (S)-Hipp-alpha-MePhe] and (S)-hippuryl-alpha-methylphenyllactic acid [(S)-Hipp-alpha-MeOPhe] were synthesized and evaluated as substrates for carboxypeptidase A (CPA) in an effort to shed further light on the catalytic mechanism of the enzyme. The rate of CPA catalyzed hydrolysis of (S)-Hipp-alpha-MePhe was reduced by 105-fold compared with that of (S)-Hipp-Phe, but the hydrolysis rate of (S)-Hipp-OPhe was lowered by only 6.8-fold by the introduction of a methyl group at the alpha-position. (R) Hipp-alpha-MePhe failed to be hydrolyzed initially, then started to undergo hydrolysis in about 2 h at a much reduced rate. The results of present study may be envisioned on the basis of the proposition that while peptide substrate is hydrolyzed via a tetrahedral transition state formed by the attack of the zinc bound water molecule at the peptide carbonyl carbon, ester hydrolysis takes the path that involves an anhydride intermediate generated by the attack of the carboxylate of Glu-270 at the ester carbonyl carbon. PMID- 10819171 TI - Discovery of a muscarinic M3 receptor antagonist with high selectivity for M3 over M2 receptors among 2-[(1S,3S)-3-sulfonylaminocyclopentyl]phenylacetamide derivatives. AB - In the course of developing a metabolically stable M3 receptor antagonist from the prototype antagonist, J-104129 (1), introduction of certain substituents into the cyclopentane ring of 1 was found to be effective not only in improving metabolic stability but also in greatly enhancing the subtype selectivity. Among the cyclopentane analogues, sulfonamide derivatives (10f) and (10g) displayed 160 and 310-fold selectivity for M3 over M2 receptors, and both were significantly more selective than the prototype antagonist (120-fold). Subsequent derivatization of the sulfonamide series led to the highly selective M3 receptor antagonists (10h, 10i and 10j) with >490-fold selectivity for M3 over M2 receptors. Among them, p-nitrophenylsulfonamide (J-107320, 10h) exhibited 1100 fold selectivity for M3 receptors (Ki = 2.5 nM) over M2 receptors (Ki = 2800 nM) in the human muscarinic receptor binding assay using [3H]-NMS as a radio ligand. PMID- 10819172 TI - Design and synthesis of novel antimicrobial pseudopeptides with selective membrane-perturbation activity. AB - By incorporating carbamate bond(s) into a cytolytic peptide, novel pseudopeptides with potent antibacterial activity and low hemolytic activity were synthesized. Circular dichroism spectra suggested that the incorporation of carbamate bond(s) decrease the alpha helical conformation of the peptide in lipid membrane circumstances, which must be regarded as a major factor for the separation of antibacterial activity from cytotoxic activity for mammalian cell. Experiments in which dye was released from vesicles indicated that the potent antibacterial activity and low hemolytic activity of the pseudopeptides must be due to their great lipid membrane selectivity. The present result suggest that backbone modifications can be a great tool for developing pseudopeptides with improved biological activity and bioavailability from cytolytic peptides. PMID- 10819173 TI - Copper contamination in paddy soils irrigated with wastewater. AB - Copper (Cu) contamination was investigated in paddy soils where Cu-rich wastewater (12 mg Cu/l) was used for irrigation. The results showed that Cu contamination increased the soil Cu content from 17.0 mg Cu/kg in the non wastewater irrigated soils (NWIS) to 101.2 mg Cu/kg in the wastewater irrigated soils (WIS), and Cu accumulated mostly in the surface layer (0-10 cm) of the paddy soil. The average Cu contents in brown rice, rice hull and rice straw from NWIS were 1.4, 7.3 and 14.5 mg Cu/kg, while those from WIS were 15.5, 133.2, and 101.4 mg Cu/kg, respectively. Correlation analysis revealed that the relationship between the Cu content in the rice straw and the rice hull with the total Cu content of the soil could be described by an exponential function (R2 = 0.921 and 0.831, respectively; P <0.01). Rice plants grown in the WIS showed symptoms of black roots, less effective tiller, etc. Subsequently, the rice yield decreased by 18-25%, compared with that grown in NWIS. PMID- 10819174 TI - Oxygen deficit determinations for a major river in eastern Hong Kong, China. AB - Determination of oxygen deficit in the Hong Kong Shing-Mun River was based on the oxygen uptake by water, algal respiration and river sediment and the oxygen supplied to the river by surface reaeration and algal photosynthesis. A systematic study was conducted to examine the effect of water temperature, flow velocity and water depth on both the oxygen demands and the oxygen supplies. The oxygen budget of a water column in a selected section of the river was modeled. The results of the oxygen budget analysis showed that when water temperature was around 10 degrees C (the lowest temperature in the river), no deficit was observed. When water temperature was 10 degrees C to 20 degrees C, a small oxygen deficit appeared, especially in the deeper water. At the highest water temperature (30 degrees C), the oxygen deficit was maximal, -6.84 g O2/m2/day, in the night-time during the Spring tide period. PMID- 10819175 TI - Effects of a copper smelter on a grassland community in the Puchuncavi Valley, Chile. AB - A grassland formation has been subjected to pollution generated by the Ventanas copper smelter since 1964 (Puchuncavi Valley, central zone of Chile) with extensive damage to local vegetation and important changes in soil characteristics. The aims of the study were (1) to detect soil parameters that best explain changes observed in plant species richness and abundance and (2) to determine if pollution-derived stresses have also affected regeneration capabilities of plant communities from the soil seed bank. The grassland was quantitatively analysed in terms of physicochemical soil characteristics, plant species diversity and abundance, and soil seed bank species composition and abundance. Results showed that a decrease in total soil nitrogen explained 13% of the changes detected in plant abundance while soil pH and 0.05 M EDTA extractable copper explained 10% and 7%, respectively, of the vegetation change. It was also found that the pollution has already affected plant species regeneration capabilities from the soil seed bank and the microsite distribution of the seeds in soils. PMID- 10819176 TI - Acid rain, storm period chemistry and their potential impact on stream communities in Hong Kong. AB - Hong Kong experiences acid deposition, however, little is known about the potential impact upon aquatic ecosystems. In a small drainage basin observations reveal that despite acid rain runoff, both baseflow and stormflow, was close to neutral in terms of pH. During storm events chemical analysis reveals that calcium (Ca) concentrations tended to rise. It also appears that the input of acid rain may increase aluminium (Al) levels in the stream. Due to the increased levels of Ca and only slight changes in pH acid deposition may not be generating problems in this stream. The presence of mayflies reported elsewhere may further support the results of the chemical study. PMID- 10819177 TI - Fractionation and composition of colloidal and suspended particulate materials in rivers. AB - The association of pollutants (nutrients, heavy metals and organic compounds) with colloidal and suspended particle matter (SPM) plays a dominant role in determining their transport, fate, biogeochemistry, bioavailability and toxicity in natural waters. A scheme for the fractionation and composition of colloidal and SPM from river waters has been tested. All four separation methods, i.e. sieving, continuous flow centrifugation, tangential flow filtration, sedimentation field-flow fractionation, were for the first time used to separate five size particulate fractions from river. Significant (gram) amounts of colloidal material (<1 microm) in three size ranges, nominally 1-0.2, 0.2-0.006 and 0.006-0.003 microm were obtained. The separation scheme was able to process large samples (100 l), within reasonable times (1 day) and the apparatus was portable. The aquatic colloid size was also characterized with high resolution by using sedimentation field-flow fractionation technique. The mass-based particle size distribution for the water sample showed a broad size distribution between 0.05 and 0.4 microm with the maximum around 0.14 microm. There was a systematic increase in the content of organic carbon (estimated by loss on ignition), Mg, Ca, Na, Cu and Zn with decreasing particle size, highlighting the importance of the colloidal (<1 microm) fraction. It was concluded that the colloidal Cu and Zn concentrations in rivers might be much higher than those reported before. PMID- 10819178 TI - Intrasite sampling of Hong Kong soils contaminated by Caesium-137. AB - Previous measurements of soil contamination by Caesium-137 (137Cs) in Hong Kong have been used both to estimate background levels prior to the construction of the Guangdong Nuclear Power Station (GNPS) at Daya Bay and to evaluate health hazards arising from the radionuclide. These measurements are reviewed and contrasted with recent advances in understanding of 137Cs distribution in soil. Preliminary research findings are used to illustrate the microscale variability of 137Cs in the Hong Kong environment and to suggest intrasite sampling methods for establishing suitable reference values. PMID- 10819179 TI - Use of protein phosphatase inhibition assay to detect microcystins in Donghu Lake and a fish pond in China. AB - Seasonal variations in the level of total microcystins in water samples collected from Donghu Lake and a fish pond in Wuhan, China, were studied between March 1995 and February 1996 using a protein phosphatase inhibition assay involving a radioactive 32P-labelled substrate. The assay is highly reliable and repeatable, and is probably the most sensitive assay for microcystin detection to date. Results of the survey indicated the presence of microcystins in the water samples, and the concentration of microcystins appeared to be related to the degree of eutrophication and water temperature. There is also a correlative relationship between the quantity of microcystins and the abundance of microcystin-producing cyanobacteria (Anabaena and Oscillatoria) in the water bodies over a year cycle. In the present study, the positive detection of microcystins in water bodies having no signs of algal bloom warns of considerable potential threat of these waters to public health. PMID- 10819180 TI - Environmental epidemiology forward. AB - Environmental epidemiology is the specialized aspect of public health science that studies human health risk from environmental hazards. It is rises largely upon a foundation of public health surveillance, and relies heavily upon analyses of data for small areas and sparse population groups. To a degree, environmental epidemiology is assigned the role of discerning very subtle human health impacts, or discerning early evidence of a tragic sequence. In that context, environmental epidemiology has a substantial public education and risk communication role. Environmental epidemiology will be greatly advanced as effective biological markers of exposure and precursor health effects are developed. At this point in time, statistical methods are in place to monitor population-level disease rates in high-risk populations for early risk identification and sentinel event recognition. Advances in geographic methods have provided a boon to the discipline by advantaging spatial studies. These advances in the discipline still need further refinement and pilot experiences. The inclusion of environmental epidemiological considerations with instances of proposed industrial expansion, hazardous waste management, and contamination remediation is heartily recommended. PMID- 10819181 TI - Development of a risk assessment methodology for evaluating potential impacts associated with contaminated mud disposal in the marine environment. AB - In order to assess impacts associated with disposal of contaminated mud arising from Hong Kong's dredging and reclamation projects, a methodology has been formulated to determine the level of risk posed by consumption of seafood/marine prey species to humans and to the Chinese White Dolphin (Sousa chinensis). This methodology improves on previously used techniques by incorporating risks for organic contaminants, accounting for doses from sources other than seafood, and incorporating additional local knowledge on Sousa chinensis behaviour. It thus represents an advance in risk assessment techniques and a new integration of risk assessment and monitoring in environmental management. PMID- 10819182 TI - The use of environmental risk assessment methodologies for an indoor air quality investigation. AB - The authors of this paper chose several target compounds that have been found in average US homes, applied the current United States Environmental Protection Agency (USEPA) Superfund risk assessment methodologies to indoor air quality, and produced risk numbers for hazard quotients and predicted increases in incidence of cancer which would be unacceptable at US hazardous waste sites. The calculations were made for the average child and adult with USEPA default exposure values. Calculations were also made for a worst case scenario using maximum concentrations and exposure estimates defined by the USEPA as describing the reasonable exposure (RME). Significant cancer risks and non-cancer hazard quotients were predicted. PMID- 10819183 TI - Risk assessment approaches for ecosystem responses to transient pollution events in urban receiving waters. AB - Alternative risk assessment approaches are reviewed for the evaluation of the ecological status and health of urban receiving waters subject to intermittent pollution events. Performance-based criteria founded on exceedance probabilities and related to the end-of-pipe discharge of chemical-specific substances comprise the conventional basis for setting regulatory standards in both North America and Europe. The difficulties and limitations of this approach, particularly in identifying realistic chronic, sub-lethal toxic risks arising from complex effluents are discussed. The potential role of Toxicity Based Criteria (TBC) for setting ecological consent limits for stormwater effluents is considered and the capabilities and limitations of Direct Toxicity Assessment (DTA) are identified. The inability of DTA procedures to satisfactorily evaluate chronic, sub-lethal risks has led to increasing interest in the potential use of in-situ biomarker techniques for the fingerprinting of stress-response properties as a means of diagnosing risk assessment for integrated urban runoff management. PMID- 10819184 TI - Human exposure to respirable suspended particulate and airborne lead in different roadside microenvironments. AB - The aim of this study is to evaluate the particulate air pollution in selected roadside microenvironments of Hong Kong through an intensive field study dated from January 1997 to February 1997. The study employed the microenvironment monitoring technique to access the exposure of pedestrians to respirable suspended particulate and airborne lead (Pb) at heavily trafficked roadsides. A total of 62 roadside sites in 14 districts covering the most urbanized and densely populated areas were selected. It was found that pedestrians were exposed to a 24 h average of respirable suspended particulate, PM10, and airborne Pb (APb), typically ranged from 25.56 to 337.40 microg/m3 and 70.71 to 285.71 ng/m3, respectively. The average PM10 concentrations at different roadside microenvironments corresponding to urban residential, urban commercial, urban industrial and new town areas were 91.84, 129.08, 83.83, and 118.89 microg/m3 respectively. The corresponding values for APb were 130.01, 143.40, 127.40 and 173.17 ng/m3, respectively. It was found that measurement at EPD nearby rooftop monitoring stations might not reflect the actual roadside PM10 exposure. Most APb field study data was significantly higher than the nearby fixed station data. PMID- 10819185 TI - Trace metals in mussels from mariculture zones, Hong Kong. AB - In 1997, concentrations of cadmium (Cd), chromium (Cr), copper (Cu), lead (Pb) and nickel (Ni) were analysed in green-lipped mussels (Perna viridis) from three mariculture zones located in the north-east (Kat O), south (Lo Tik Wan) and to the west (Ma Wan) of Hong Kong. Spatial differences in the concentration of metals were found, chromium and copper were higher at Ma Wan and Lo Tik Wan compared to Kat O in the north-east. In contrast, the highest levels of lead (mean = 4.37 microg/g dry wt) were recorded at Kat O. There were no differences in the level of nickel between the study sites. A comparison of the metal concentrations in mussels with the results of a previous study seven years before, in 1990, showed a twofold increase in the mean levels of cadmium for all three sites. However, levels of the other metals in 1997 were lower by 12-32% for chromium, 32-39% for copper and 24-25% for nickel. The greatest reductions were recorded for lead: Kat O (39%), Ma Wan (51%) and Lo Tik Wan (75%). This may be related to the introduction of lead-free petrol in 1991. Despite the apparent reduction in some heavy metal bioaccumulation between 1990 and 1997, from a public health risk perspective, the data suggest a continued need for monitoring of heavy metals in mussels from mariculture zones. PMID- 10819186 TI - Indoor and outdoor air quality investigation at schools in Hong Kong. AB - Five classrooms in Hong Kong (HK), air-conditioned or ceiling fans ventilated, were chosen for investigation of indoor and outdoor air quality. Parameters such as temperature, relative humidity (RH), carbon dioxide (CO2), sulphur dioxide (SO2), nitric oxide (NO), nitrogen dioxide (NO2), respirable particulate matter (PM10), formaldehyde (HCHO), and total bacteria counts were monitored indoors and outdoors simultaneously. The average respirable particulate matter concentrations were higher than the HK Objective, and the maximum indoor PM10 level exceeded 1000 microg/m3. Indoor CO2 concentrations often exceeded 1000 microl/l in air conditioning and ceiling fan classrooms, indicating inadequate ventilation. Maximum indoor CO2 level reached 5900 microl/l during class at the classroom with cooling tower ventilation. Increasing the rate of ventilation or implementation of breaks between classes is recommended to alleviate the high CO2 level. Other pollution parameters measured in this study complied with the standards. The two most important classroom air quality problems in Hong Kong were PM10 and CO2 levels. PMID- 10819187 TI - Effects of arsenic on blast transformation and DNA synthesis of human blood lymphocytes. AB - Effects of inorganic arsenicals on DNA synthesis in unsensitized human blood lymphocytes were biphasic: the chemicals at very low concentrations enhanced blast transformation and DNA synthesis, whereas higher concentrations inhibited the transformation and DNA synthesis. The concentrations of arsenicals at which the maximum stimulating effect was found were 1 x 10(-5) M, 1 x 10(-6) M or 2 x 10(-6) M, and 0.8 x 10(-6) M or 1 x 10(-6) M for sodium arsenite exposure of 1 h, 3 days and 6 days, respectively; for sodium arsenate, 1 x 10(-5) M, 1 x 10(-5) M, and 2 x 10(-6) M or 5 x 10(-6) M, respectively. Arsenicals must be present for the entire 6 days culture period to produce maximum stimulation of blast transformation of human lymphocytes. The longer exposure of the lymphocytes to arsenicals, the lower the concentrations of arsenicals at which the maximum stimulating effect was found. The stimulating effect of trivalent arsenic (sodium arsenite) was stronger than pentavalent arsenic (sodium arsenate). PMID- 10819188 TI - Soil contamination with radionuclides and potential remediation. AB - Soils contaminated with radionuclides, particularly 137Cs and 90Sr, pose a long term radiation hazard to human health through exposure via the foodchain and other pathways. Remediation of radionuclide-contaminated soils has become increasingly important. Removal of the contaminated surface soil (often up to 40 cm) or immobilization of radionuclides in soils by applying mineral and chemical amendments are physically difficult and not likely cost-effective in practicality. Reducing plant uptake of radionuclides, especially 137CS and 90Sr by competitive cations contained in chemical fertilizers has the general advantage in large scale, low-level contamination incidents on arable land, and has been widely practiced in central and Western Europe after the Chernobyl accident. Phytoextraction of radionuclides by specific plant species from contaminated sites has rapidly stimulated interest among industrialists as well as academics, and is considered to be a promising bio-remediation method. This paper examines the existing remediation approaches and discusses phytoextraction of radionuclides from contaminated soils in detail. PMID- 10819189 TI - Genotoxicity of surface water samples from Meiliang Bay, Taihu Lake, Eastern China. AB - Taihu Lake is the third largest freshwater lake in China. Taihu Basin is one of the most densely populated and urbanized areas in this country. This area provides 15% of the GDP. Meiliang Bay is located in the north part of the Lake. It provides the municipal water source for Wuxi City. Some parts of the lake have been found to be highly polluted due to eutrophication for over a decade. Surface water (0-0.5 m) samples were collected from the Meiliang Bay by the aid of Global Position System (GPS) for positioning. Water samples were concentrated 5000 times with XAD-2 resin columns. A reverse mutation test using histidine-dependent Salmonella typhimurium strains was employed to assay the genotoxicity of the samples. The results showed that the sample from position 6 had the highest genotoxicity either in the case of activating with eucaryotic S9 system or without S9. The genotoxic effect included, at least, two different molecular mechanisms: nucleotide point substitution on DNA molecules and reading frame shifting caused by nucleotide insertion or deletion. The genotoxicity of the water body in Meiliang Bay, Taihu Lake should be kept in close monitoring. PMID- 10819190 TI - Effect of applying chemical fertilizers on forms of lead and cadmium in red soil. AB - A three-month incubation study was undertaken to examine the influence of N, P and K on the various forms (soluble plus exchangeable (SE), weakly specifically adsorbed (WSA), Fe-Mn oxides bound (OX), organic matter complexed (OM) and residual fractions (RES)) of lead (Pb) and cadmium (Cd) in a red soil. Application of urea at the rate of 200 mg N/kg significantly lowered the SE fraction, but raised the WSA or OX fraction of both metals. Supply of 80 mg P/kg caused a decrease in the SE fraction of the two metals. The WSA fraction of Pb was reduced, whereas that of Cd increased by adding P. However, addition of 100 mg K/kg led to an increase in the SE fraction, but a decrease in the WSA fraction of Pb and Cd. Applying chemical fertilizers had no significant consistent influences on the other fractions of metals. These findings suggest that in heavy metal contaminated red soil, applying fertilizers does not only provide plant nutrients, but may also change the speciations and thus biovailability of heavy metals. PMID- 10819191 TI - Effects of metabolites of benzo(a)pyrene on unschedule DNA synthesis in BALB/3T3 cell line. AB - In order to explore the damage from metabolites of benzo(a)pyrene on DNA of mammalian cells, the effects of four metabolites of benzo(a)pyrene (anti-BPDE, syn-BPDE, 3-OH-BP and 9-OH-BP) on synthesis of DNA and unschedule DNA synthesis (UDS) in BALB/3T3 cells were assayed, by methods of single-labeling and double labeling. The results showed that all of the four agents were able to increase the synthesis of DNA, but only three of them (apart from syn-BPDE) induced UDS in BALB/3T3 cells. The above indicates that the metabolites of benzo(a)pyrene are able to damage DNA in BALB/3T3 cells, and that this effect may be relative to the sterical structure of metabolites of benzo(a)pyrene. PMID- 10819192 TI - Study on the cytotoxicity of microcystin-LR on cultured cells. AB - The toxicity of purified blue-green algal toxin, microcystin-LR, on permanent cell lines KB, NIH/3T3, H-4-II-E, HeLa, Vero, Hep G2, Caco-2 and HL-60 was studied. Assessment of cell viability using colorimetric 3-(4,5-dimethylthiazol-2 yl)-2,5-diphenyltetrazolium bromide (MTT) assays indicated that purified microcystin-LR induced toxic effect on KB and H-4-II-E cell lines after 96 h incubation at toxin concentrations greater than 18.75 microg/ml. KB cell line was selected for further study when reproducibility, consistency and sensitivity were considered. Significant amounts of lactate dehydrogenase (LDH) were released from KB cells when incubation durations were 72 and 96 h with toxin concentrations of 18.75 microg/ml and higher. Although previous studies suggested that microcystin LR had no cytotoxic effect on permanent cell lines, LDH release assay performed on KB cells indicated that exposure to microcystin-LR could result in damage to the cell membrane. PMID- 10819193 TI - Degradation pathway of persistent branched fatty acids in natural anaerobic ecosystem. AB - Branched fatty acids (BAFs) in industrial effluents are often persistent in biological wastewater treatment systems and end up as organic contaminants in the water environment. In this study, degradation of eight characteristic BAFs in a natural anaerobic ecosystem of an eutrophic river sediment was studied in vitro by enrichment culture techniques. The anaerobic consortium, comprising of BFA degrading and methane-producing genera, degraded BFAs with a tertiary carbon through beta-oxidation followed by methanogenesis mechanisms. The consortium could not degrade BFAs with a quaternary carbon. The degree of branching at the alpha or beta position along the carbon chain interfered with the beta-oxidation mechanisms, and hence affected the degradability of the compound. PMID- 10819194 TI - Studies on thallium toxicity, its tissue distribution and histopathological effects in rats. AB - The distribution of thallium (Tl) in the body and its toxic effect on the histology and function of the liver and kidney of rats after Tl administration were investigated using biochemical and histopathological assays. Male albino rats exhibited a markedly dose-dependent increase in the serum levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) at 16 h after an intraperitoneal injection of 30, 60 or 120 mg/kg Tl. The serum level of creatinine in the rats injected with 30 mg/kg Tl, elevated significantly after 4 days of administration. The distribution of Tl in the tissues of intoxicated rats was uneven. The content of Tl was found to be highest in the kidney, followed by ileum, stomach and liver. Histological examination demonstrated frequent occurrence of hepatocyte necrosis and vacuolation in the liver and pathological changes of renal tubules in the treated rats. PMID- 10819195 TI - Soil solution Zn and pH dynamics in non-rhizosphere soil and in the rhizosphere of Thlaspi caerulescens grown in a Zn/Cd-contaminated soil. AB - Temporal changes in soil solution properties and metal speciation were studied in non-rhizosphere soil and in the rhizosphere of the hyperaccumulator Thlaspi caerulescens J. & C. Presl (population from Prayon, Belgium) grown in a Zn- and Cd-contaminated soil. This paper focuses on soil solution Zn and pH dynamics during phytoextraction. The concentration of Zn in both non-rhizosphere and rhizosphere soil solutions decreased from 23 mg/l at the beginning to 2 mg/l at the end of the experiment (84 days after transplanting of seedlings), mainly due to chemical sorption. There was no significant difference in overall Zn concentration between the planted and the unplanted soil solutions (P > 0.05). Soil solution pH decreased initially and then increased slightly in both planted and unplanted soil zones. From 60 to 84 days after transplanting, the pH of the rhizosphere soil solution was higher than that of non-rhizosphere soil solution (P<0.05). Zn uptake by the hyperaccumulator plants was 8.8 mg per pot (each containing 1 kg oven-dry soil) on average. The data indicate that the potential of T. caerulescens to remove Zn from contaminated soil may not be related to acidification of the rhizosphere. PMID- 10819196 TI - Effects of chronic lead exposure on short-term and long-term depression in area CA1 of the rat hippocampus in vivo. AB - Chronic developmental lead (Pb) exposure to the rat has been reported to impair the long-term potentiation (LTP) in area CA1 and DG of the hippocampus. The present study was performed to investigate the effects of chronic Pb exposure on homosynaptic short-term depression (STD) and long-term depression (LTD) of population spikes (PS) in area CA1 of the rat hippocampus in vivo. Neonatal Wistar rats were exposed to Pb from parturition to weaning via the milk of dams fed with 0.2% lead acetate solution. The input/output (I/O) function, paired pulse reaction (PPR), the PS were measured in the area CA1 in response to low frequency stimulation (LFS). The results showed that the homo-STD amplitude of PS depotentiation in Pb-exposed rats (87.48 +/- 7.44%, n = 14) was less significant than that in control rats (72.34 +/- 6.05%, n = 18, P<0.05), and the homo-LTD amplitude of PS depotentiation in Pb-exposed rats (72.80 +/- 5.86%, n = 14) was even less significant than that in control rats (47.80 +/- 5.03%, n = 18, P<0.01). The results suggest that chronic Pb exposure in neonatal rats caused impairments in the STD and LTD of area CA1 of hippocampus. PMID- 10819197 TI - Protective effects of seaweeds against liver injury caused by carbon tetrachloride in rats. AB - Three species of seaweeds collected from Tung Ping Chau, Hong Kong, were screened for their hepatoprotective activity using carbon tetrachloride (CCl4)-induced liver injury in the rat as a model of chemical hepatitis. A single oral dose of 1.25 ml/kg of CCl4 was able to produce significantly elevated levels of serum glutamic pyruvic transaminase (GPT) and glutamic oxaloacetic transminase (GOT). Administration of 150, 300 and 600 mg/kg of aqueous extracts from Myagropsis myagroides, Sargassum henslowianum and S. siliquastrum, respectively, significantly reduced the CCl4-induced acute elevation in the levels of GPT and GOT in rats. The same result was also seen in the histopathological study of liver tissue. The seaweed crude extracts probably acted to protect against CCl4 induced liver injury through their antioxidant properties. PMID- 10819198 TI - Effects of 2,4-D, glyphosate and paraquat on growth, photosynthesis and chlorophyll-a synthesis of Scenedesmus quadricauda Berb 614. AB - The effects of 2,4-D, glyphosate and paraquat on growth, photosynthesis and chlorophyll-a synthesis by a freshwater green alga, Scenedesmus quadricauda Berb 614, were determined. These herbicides are the most often used in Hong Kong. Within the concentration range 0.02-200 mg/l, paraquat was more toxic than glyphosate and 2,4-D to the growth, photosynthesis and chlorophyll-a synthesis. The presence of 0.02, 0.2 or 2 mg/l of 2,4-D was not toxic to the alga. Algal growth, photosynthesis and chlorophyll-a synthesis were stimulated by the presence of low concentrations (0.02 or 0.2 and 0.02 mg/l, respectively) of 2,4-D and glyphosate. The presence of 0.02 or 0.2 mg/l of paraquat, 2 mg/l of glyphosate or 20 mg/l of 2,4-D was significantly inhibitory to the three test parameters, whereas the presence of 2 or more mg/l of paraquat, 20 or more mg/l of glyphosate or 200 mg/l of 2,4-D completely inhibited algal growth, photosynthesis and chlorophyll-a synthesis. The use of the alga as a bio indicator of herbicide contamination in freshwater environment was discussed. PMID- 10819199 TI - Effects of tri-o-cresyl phosphate on serum estrogen and progesterone concentration and ATPase activity in the shell gland of adult hens. AB - The activities of calcium-activated ATPase (Ca2+-ATPase) and calcium magnesium activated ATPase (Ca2+-Mg2+-ATPase) in the shell gland, and concentrations of 17beta-estradiol (E2) and progesterone in serum were monitored, respectively, from hens orally dosed with tri-o-cresyl phosphate (TOCP) (750 mg/kg). Treated birds were monitored daily for laying and development of delayed neurotoxicity, and activities of Ca2+-ATPase and Ca2+-Mg2+-ATPase were measured at 7 and 10 days after dosing. TOCP-treated birds manifested motor deficit by 7-9 days postdosing, while hens administered vehicle exhibited no signs of delayed neurotoxicity. Ca2+ ATPase and Ca2+-Mg2+-ATPase activities of shell glands from TOCP-dosed hens were not significantly affected (P > 0.05). The serum E2 concentration was significantly reduced in TOCP-treated hens (P < 0.01); however, progesterone levels were unaffected. PMID- 10819200 TI - Promotive effect of diethylstilbestrol on urethan-induced mouse lung tumorigenesis. AB - The effect of diethylstilbestrol (DES) on urethan-induced mouse lung tumorigenesis was assessed by a single intraperitoneal injecting of urethan (50 mg/kg) or/and multi intramuscular injecting of DES (5 or 50 mg/kg). All mice were sacrificed 18 weeks after administration, and the lung tumors were examined histopathologically. DES did not produce an elevated lung tumor response when administered alone, but it produced a statistically significant enhancement of incidence of tumors, average number of tumors, incidence of cancers and constituent ratio of malignant tumors when given in conjunction with urethan. The results indicated that DES may be a promoter in lung tumor formation. PMID- 10819201 TI - Acute toxicity of excess mercury on the photosynthetic performance of cyanobacterium, S. platensis--assessment by chlorophyll fluorescence analysis. AB - Measurement of chlorophyll fluorescence has been shown to be a rapid, non invasive, and reliable method to assess photosynthetic performance in a changing environment. In this study, acute toxicity of excess Hg on the photosynthetic performance of the cyanobacterium S. platensis, was investigated by use of chlorophyll fluorescence analysis after cells were exposed to excess Hg (up to 20 microM) for 2 h. The results determined from the fast fluorescence kinetics showed that Hg induced a significant increase in the proportion of the Q(B)-non reducing PSII reaction centers. The fluorescence parameters measured under the steady state of photosynthesis demonstrated that the increase of Hg concentration led to a decrease in the maximal efficiency of PSII photochemistry, the efficiency of excitation energy capture by the open PSII reaction centers, and the quantum yield of PSII electron transport. Mercury also resulted in a decrease in the coefficients of photochemical and non-photochemical quenching. Mercury may have an acute toxicity on cyanobacteria by inhibiting the quantum yield of photosynthesis sensitively and rapidly. Such changes occurred before any other visible damages that may be evaluated by other conventional measurements. Our results also demonstrated that chlorophyll fluorescence analysis can be used as a useful physiological tool to assess early stages of change in photosynthetic performance of algae in response to heavy metal pollution. PMID- 10819202 TI - Role of plants, mycorrhizae and phytochelators in heavy metal contaminated land remediation. AB - Phytoremediation is a site remediation strategy, which employs plants to remove non-volatile and immisible soil contents. This sustainable and inexpensive process is emerging as a viable alternative to traditional contaminated land remediation methods. To enhance phytoremediation as a viable strategy, fast growing plants with high metal uptake ability and rapid biomass gain are needed. This paper provides a brief review of studies in the area of phytoaccumulation, most of which have been carried out in Europe and the USA. Particular attention is given to the role of phytochelators in making the heavy metals bio-available to the plant and their symbionts in enhancing the uptake of bio-available heavy metals. PMID- 10819203 TI - Impacts of a mosquito selective pesticide, Bti, on the macroinvertebrates of a subtropical stream in Hong Kong. AB - To test the hypothesis that Bti was specific to mosquito larvae, the granular form of Bti was tested on commonly found Hong Kong species from four naturally occuring aquatic insect orders and one species of decapod crustacean (Neocardina serra). Because data on Bti impacts on species in tropical and subtropical countries is relatively rare, the present study was conducted in Hong Kong's New Territories. Using static acute toxicity bioassays, all mosquito larvae exposed to Bti at the recommended dosage were dead except for some dark pigmented fourth instar individuals that had stopped feeding prior to emergence and as a result, did not ingest the Bti toxin. The only non-target species killed by the Bti were some chironomid species. In 1998 Bti inoculated pools along the Tai Tan River in the New Territories of Hong Kong had significantly fewer chironomids and mosquito larvae than control pools. By January, 1999, these same Bti inoculated pools still had no mosquito larvae in them, however, chironomids were no longer significantly rarer when compared to the control ponds. Thus, it would appear, that during the 1998-99 study in which Bti was added to the study sites at weekly intervals, Bti resistant chironomid species replaced Bti sensitive species in the Bti inoculated pools. PMID- 10819204 TI - Current approaches to the revegetation and reclamation of metalliferous mine wastes. AB - Abandoned metalliferous mine wastes can result in severe pollution and have aesthetic impacts on the local environment. Use of a vegetation cover gives a cost-effective and environmentally sustainable method of stabilising and reclaiming wastes such as mine-spoils and tailings. Many characteristics of metalliferous wastes are often inimical to successful vegetation establishment, most notably phytotoxic levels of residual heavy metals, low nutrient status and poor physical structure of the substratum. Current approaches to revegetation and reclamation involve both ameliorative and adaptive strategies to allow plant establishment and encourage subsequent vegetation development. Different techniques of revegetation are available for temperate and arid, subtropical regions depending on the characteristics of the waste. These include direct seeding with commercially available plants, use of cover and barrier systems and the enhancement of natural revegetation processes. PMID- 10819205 TI - Chemical methods and phytoremediation of soil contaminated with heavy metals. AB - The effects of chemical amendments (calcium carbonate (CC), steel sludge (SS) and furnace slag (FS)) on the growth and uptake of cadmium (Cd) by wetland rice, Chinese cabbage and wheat grown in a red soil contaminated with Cd were investigated using a pot experiment. The phytoremediation of heavy metal contaminated soil with vetiver grass was also studied in a field plot experiment. Results showed that treatments with CC, SS and FS decreased Cd uptake by wetland rice, Chinese cabbage and wheat by 23-95% compared with the unamended control. Among the three amendments, FS was the most efficient at suppressing Cd uptake by the plants, probably due to its higher content of available silicon (Si). The concentrations of zinc (Zn), lead (Pb) and Cd in the shoots of vetiver grass were 42-67%, 500-1200% and 120-260% higher in contaminated plots than in control, respectively. Cadmium accumulation by vetiver shoots was 218 g Cd/ha at a soil Cd concentration of 0.33 mg Cd/kg. It is suggested that heavy metal-contaminated soil could be remediated with a combination of chemical treatments and plants. PMID- 10819206 TI - The effects of chemical remediation treatments on the extractability and speciation of cadmium and lead in contaminated soils. AB - Two rural soils contaminated by cadmium (Cd) and lead (Pb) were used to evaluate the effect of different chemical treatments on changes in speciation and extractability of Cd and Pb, and in phytoavailability to wheat. Triplicates of seven chemical treatments were tested to compare and evaluate the remediation techniques for contaminated soils using pot experiments. Treatments applied were calcium carbonate, a high quantity of phosphate salt, hog composts, iron oxide, manganese oxide, zeolite, and unamended control. Wheat (Triticum aestivum) was planted in the different amended soils for a further one month to evaluate the effectiveness of treatments on uptake of Cd and Pb by the wheat shoots. Results indicated that addition of calcium carbonate, manganese oxide, or zeolite reduces the extractability of Cd or Pb in both soils, and significantly reduce the uptake of Cd and Pb by wheat shoots. Changes in the extractability and metal sequential fractionations indicate that the exchangeable (or available) form of Cd and Pb in two soils can be transformed into unavailable forms after these amendments. PMID- 10819207 TI - Improving phosphate removal of sand infiltration system using alkaline fly ash. AB - Septic tank effluent is customarily disposed of by soil infiltration. Coarse, sandy soil such as those found in Perth, Western Australia, exhibit low attenuation capabilities for phosphate (PO4(3-)) during effluent infiltration. Amendment of such soil with different amounts of alkaline precipitator and lagoon fly ashes was investigated as a means of reducing phosphorus (P) leakage to ground water. Alkaline precipitator fly ash possessed the highest P sorption capacity in terms of its Langmuir and Freundlich isotherm parameters during initial batch tests. The test materials were repeatedly contacted with fresh PO4(3-) solutions over 90 contacting cycles to gain a better indication of long term P sorption capability. Again, precipitator fly ash exhibited higher P sorption capacity than lagoon fly ash and Spearwood sand. Column studies assessed the influence of various application rates of alkaline precipitator and lagoon fly ashes on the P removal of septic tank effluent. Septic tank effluent was applied at the rate of 4 cm/day to the column for 12 weeks. Concentrations of P were monitored in the column effluent. All the fly ash columns were more efficient in reducing P migration compared to the sand column. Increased levels of fly ash in the soil columns resulted in increased P attenuation. Lagoon fly ash was inferior to precipitator fly ash for P removal; high application rates of fly ash caused clogging of the infiltration bed apparently due to their lower permeability. It is reasoned that 5-15% precipitator fly ash, and less than 30% lagoon fly ash could be added to coarse sands to produce an infiltration bed, which would result in a better quality effluent than can be obtained with untreated sand alone. PMID- 10819208 TI - Performance of different microalgal species in removing nickel and zinc from industrial wastewater. AB - A series of batch experiments was conducted to compare the ability of 11 microalgal species of the same cell density in removing nickel (Ni) and zinc (Zn) from synthetic wastewater. These included Chlorella vulgaris (commercially available), Chlorella sorokiniana and Scenedesmus quadricauda (isolates from polluted water of Wuhan, China), and eight different isolates from Hong Kong. The Wuhan isolate of Scenedesmus removed most Ni, probably due to its large biomass. Nickel concentration was reduced from an initial 30 to 0.9 mg/l after 5 min (97% Ni removal), and further declined to 0.4 mg/l after 90 min of treatment. In wastewater containing 30 mg/l Ni and 30 mg/l Zn, more than 98%, Ni and Zn were removed simultaneously at the end of 5 min treatment, indicating that the presence of Zn in wastewater did not affect Ni removal by this Scenedesmus isolate. The second most effective species for Ni removal was an isolate, tentatively identified as Chlorella miniata, Ni concentration was reduced to 10 mg/l after 90 min, and was only slightly interfered by the presence of Zn. In terms of metal removal per unit biomass or unit surface area of algal cells, C. miniata was the best species in removing Ni and Zn. At the other extreme, one Hong Kong isolate (Synechocystis sp.) did not remove any Ni and only achieved 40% Zn removal. Performance of the other isolates was comparable with the commercial C. vulgaris, less than 50% Ni was removed after 5 h of treatment and Ni removal was significantly reduced by the presence of Zn. All algae tested were found to be viable, showing these 11 species could tolerate a mixture of 30 mg/l Ni and 30 mg/l Zn in wastewater. PMID- 10819209 TI - Strategies for sustainable woodland on contaminated soils. AB - Extensive in situ reclamation treatment technologies are appropriate for a large proportion of contaminated land in place of total removal or complete containment of soil. In this paper, initial results are presented of site descriptions, tree survival and metal uptake patterns from two field planting trials on a highly industrially contaminated site adjacent to a metal refinery and on old sanitary landfill sites. Survival rate was high in both trials but factors besides heavy metals were particularly significant. Uptake patterns of metals into foliage and woody tissues were variable, with substantial uptake in some species and clones supporting the findings of earlier pot experiments. It is argued that there is sufficient evidence to consider the use of trees in reclamation as part of a realistic, integrated, low-cost, ecologically-sound and sustainable reclamation strategy for contaminated land. This is an opportunity to bring a large number of brownfield sites into productive use, which otherwise would be prohibitively expensive to restore. PMID- 10819210 TI - Synthetic zeolites as amendments for sewage sludge-based compost. AB - The effects of incorporating a synthetic zeolite (Zeolite P) in a range of concentrations (0.1-1.0 w:w) into an experimental horticultural compost, derived from sewage sludge, have been investigated. The impact of zeolite treatment on time-related changes of the labile zinc, copper, iron and manganese pools within the compost was compared to lime incorporation (5% w:w) and to a proprietary unamended peat-based compost. Addition of 0.5% and 1.0% zeolite significantly reduced labile zinc over a 90 day period. The highest zeolite treatment was more effective than liming; 0.5% zeolite was as effective as lime. Plant growth trials measuring transfer of metals to ryegrass (Lolium perenne L. cv Elka) in successive harvests demonstrated that both 1.0% zeolite and 5% lime treatment caused significant reduction in total metal transfer from soil-plant over a 116 day growth period. It is concluded that the use of synthetic zeolite as an amendment for compost of this type significantly reduces potential for soil metal mobility and soil-plant transfer. PMID- 10819211 TI - Ionic strength effects in biosorption of metals by marine algae. AB - Biosorption, the passive accumulation of metals by biomass, can be used as a cost effective process for the treatment of metal polluted industrial effluents. The green alga Ulva fascia and the brown seaweeds Sargassum hemiphyllum, Petalonia fascia, and Colpomenia sinuosa were characterized in terms of their number of binding sites, their charge density and intrinsic proton binding constant (pKa) using pH titrations at different ionic strengths. The determined number of binding sites decreased in the order Petalonia > or = Sargassum > Colpomenia > Ulva. Due to their high number of binding sites Sargassum and Petalonia are most promising for biosorption applications. The decrease of proton binding with increasing ionic strength and pH as well as the increase of Cu and Ni binding with increasing pH and decreasing ionic strength could be described by the Donnan model in conjunction with an ion exchange biosorption isotherm. PMID- 10819212 TI - Removal of heavy metals from anaerobically digested sewage sludge by isolated indigenous iron-oxidizing bacteria. AB - The removal of heavy metals (Cr, Cu, Zn, Ni and Pb) from anaerobically digested sludge from the Yuen Long wastewater treatment plant, Hong Kong, has been studied in a batch system using isolated indigenous iron-oxidizing bacteria. The inoculation of indigenous iron-oxidizing bacteria and the addition of FeSO4 accelerated the solubilization of Cr, Cu, Zn, Ni and Pb from the sludge. pH of the sludge decreased with an increase in Fe2+ concentrations and reached a low pH of 2-2.5 for treatments receiving both bacterial inoculation and FeSO4. After 16 days of bioleaching, the following heavy metal removal efficiencies were obtained: Cr 55.3%, Cu 91.5%, Zn 83.3%, Ni 54.4%, and Pb 16.2%. In contrast, only 2.6% of Cr, 42.9% of Cu, 72.1% of Zn, 22.8% of Ni and 0.56% of Pb were extracted from the control without the bacterial inoculation and addition of FeSO4. The residual heavy metal content in the leached sludge was acceptable for unrestricted use for agriculture. The experimental results confirmed the effectiveness of using the isolated iron-oxidizing bacteria for the removal of heavy metals from sewage sludge. PMID- 10819213 TI - Linking ecological and ecotoxicological techniques to support river rehabilitation. AB - Human activities in river catchments interfere with natural fluxes of water and materials. Diffuse inputs and point-sources of toxicants have modified the ecological state of riverine communities considerably, and sanitation schemes are now under development for various rivers. To improve analysis, monitoring and prospecting the role of toxicants in river ecosystems a review of the available methods is undertaken. Ecotoxicological techniques are discussed in relation to basic ecological principles that are thought to regulate the functioning of communities. The response to toxicants among species is highly diverse and therefore the choice of test species (e.g. of typical riverine insects as caddisflies or mayflies) is critical, as it is the use of test-batteries. Long term exposure may lead to developmental disturbances that may be assessed through morphometric techniques like analysis of asymmetry. Multi-generation exposure, although rarely studied, provides a useful insight into the genetic consequences of pollution. Selection for tolerant species or varieties has been experimentally assessed for smaller organisms such as insects, micro-algae, and bacteria. There is also perspective for multivariate analysis of species distribution in relation to pollutant exposure. Furthermore, a system approach to benthic ecology and sediment testing is needed. Such an approach reflects the strong linkage of ecological and ecotoxicological processes. Toxicants are transformed by biological activity; in some cases this alleviates toxicant stress, but in other cases degradation products are toxic as well. The risk of transformation to mutagenic products in the environment is indicated. The re-assessment of some of the classical ecotoxicological techniques is needed to adequately fulfil the needs of ecological recovery programs. To this purpose integration of ecotoxicological and ecological tools is needed. PMID- 10819214 TI - Separation of toxaphene by high resolution gas chromatography. AB - A summary is given about the separation of toxaphenes by high resolution gas chromatography (HRGC). The suitability of different stationary phases for isomer and/or enantiomer-selective separations is compared. Multidimensional and tandem techniques are also presented. In addition, problems caused by thermal degradation in the injector and on the column are addressed. Furthermore, a brief survey is included about detection methods such as electron capture detection and different mass spectrometric methods. PMID- 10819215 TI - Relative retention times of chlorinated monoterpenes. AB - GC retention times for 67 chlorinated monoterpenes, belonging to the technical pesticide Toxaphene have been measured under the same conditions and relative retention times have been determined. They consisted of 45 chlorinated bornanes, 13 chlorinated bornenes, 8 chlorinated dihydrocamphenes and 1 chlorinated camphene. Useful correlations between structure and retention time were found. Similar changes in structure for different compounds lead to similar changes in retention times. These correlations allow to predict retention times for yet unknown Toxaphene congeners. PMID- 10819216 TI - Discrimination and thermal degradation of toxaphene compounds in capillary gas chromatography when using split/splitless and on-column injection. AB - Technical toxaphene and a 22-component Reference Mixture were analyzed using capillary gas chromatography with split/splitless injection (SSL) and on-column injection (OC). In both techniques, electron-capture, negative ionization mass spectrometry (ECNI-MS) was used for detection of chlorobornanes, chlorocamphenes and related compounds. Significant discrimination of highly chlorinated congeners was observed as a result of incomplete transfer of these compounds from the vaporizer to the analytical column when using SSL. This resulted in a much lower response for nona- and decachloro congeners than when using OC. In addition, several toxaphene components, especially the chlorobornanes with gem dichloro substitution on the six-member carbon ring, undergo thermal degradation when using SSL. Some of these congeners are major components of technical toxaphene, but generally are not present, except at low concentrations, in environmental and biological samples. Therefore, technical toxaphene may be discriminated and/or degraded differently than toxaphene compounds in environmental samples when using SSL. This results in significant bias of the quantitative data when using the technical material as a reference. OC suffers much less from these deficiencies and, therefore, is a preferable technique for toxaphene analysis. PMID- 10819217 TI - The application of standard methods for the determination of toxaphene in environmental media. AB - In the United States, it is necessary to analyze for toxaphene using approved, validated methods acceptable to the regulatory agencies. As a result of an interlaboratory study and technical exchanges among the US EPA Region IV (EPA), the State of Georgia Environmental Protection Division (EPD), and Hercules Incorporated, guidelines for the application of SW-846 Method 8080 were developed. Results of analyses for sludge, soil, and water samples agreed within a percent relative standard deviation (% RSD) range of 6.9-20%. Through continued technical interchanges, guidance for the application of SW-846 Method 8081 has been developed. The results of analyses of fourteen split samples of soil and sediment produced a percent relative standard deviation that ranged from 1.6% to 127%, with an average of 38%. When two unusually divergent results were removed from consideration, the average % RSD reduced to 26%. The results of analyses of split samples show agreement between the EPA laboratory and a Hercules contract laboratory. Therefore, the guidance has achieved its purpose of producing agreement among data from different laboratories so that data reviewers may have assurance that the analytical method has been applied correctly and consistently for the determination of toxaphene in environmental samples. PMID- 10819218 TI - The determination of toxaphene in environmental samples by negative ion electron capture high resolution mass spectrometry. AB - A method is described for the analysis of toxaphene in environmental samples by high resolution gas chromatography high resolution mass spectrometry in selected ion electron capture negative ion mode. Toxaphene is detected by monitoring [M Cl]- fragments ions of chlorobornanes (Cl6-Cl10) and chlorobornenes (Cl6-Cl7) using formal ion ratio criteria. The method shows high sensitivity and excellent specificity for the detection of chlorobornanes with negligible interference from of a wide range of common environmental organochlorines including chlordanes, PCBs and PCB-oxygen adducts. The analytical scheme is applicable to sediment and biological tissue samples and typically provide sample detection limits of less than 0.2 ng/g technical toxaphene for a 10 g sample. PMID- 10819219 TI - Toxaphene in standard solutions and cleaned biota extracts--results of the first QUASIMEME interlaboratory studies. Quality Assurance of Information for Marine Environmental Monitoring in Europe. AB - Two interlaboratory studies on individual toxaphene congeners have been organised by the project Quality Assurance of Information for Marine Environmental Monitoring in Europe (QUASIMEME). Fifteen laboratories analysed two standard solutions in the first study and 13 laboratories analysed a standard solution and two cleaned biota extracts in the second study. The coefficients of variation obtained for the standard solutions were 6-21% and for the cleaned extracts 16 39%. Although the results were comparable to those of other studies, further improvement in the level of agreement between the participating laboratories was considered possible. PMID- 10819220 TI - Enantioselective determination of persistent and partly degradable toxaphene congeners in high trophic level biota. AB - Enantiomer separation of chiral toxaphene components in biological samples was studied by application of different chiral stationary phases based on modified cyclodextrins. Several pairs of enantiomers were resolved on permethylated beta cyclodextrin (beta-PMCD), among them 2-endo,3-exo,5-endo,6-exo,8,8,9,10 octachlorobornane (B8-1412), which was not enantiomerically resolved on tert butyldimethylsilylated beta-cyclodextrin (beta-BSCD). The latter column was applied to determine the enantiomer ratios (ERs) of 2-endo,3-exo,5-endo,6 exo,8,8,10,10-octachlorobornane (B8-1413 or P-26) in brain tissue of three seal species. The ER of B8-1413 (P-26) in brain was virtually racemic as well as those of the two persistent and chiral components of technical chlordane, 1 exo,2,2,4,5,6,7,8,8-octachloro-3a,4,7,7a-tetrahydro-4,7-metha noindane (trans nonachlor III or MC 6) and 1-exo,2-endo,3-exo,4,5,6,8,8-octachloro-3a,7,7a tetrahydro-4,7- methanoindane (U82). In contrast, B8-1412 and 2 exo,5,5,8,9,9,10,10-octachlorobornane (B8-2229 or P-44) were significantly enantiomerically enriched in several samples of high trophic level biota. 2,2,5,5,8,9,9,10,10-Nonachlorobornane (B9-1025 or P-62), a chlorobornane metabolisable by seals and the presumable precursor of B8-2229 (P-44), was also enantiomerically enriched in seal blubber. These results confirm the assumption that some less persistent toxaphene components may be significantly degraded in biological samples. Enantioselective gas chromatography provides the information that such a degradation is happening by the characteristic change of the ratio of the two enantiomers in the respective tissues. PMID- 10819221 TI - Total toxaphene and specific congeners in fish from the Yukon, Canada. AB - Toxaphene is one of the major persistent organic pollutants with global environmental impacts. We have measured total toxaphene and specific congeners concentrations in 19 fish samples collected from the Yukon, Canada using gas chromatography coupled to ion trap MS/MS. The total toxaphene concentrations ranged from 42 to 242 ng/g (mean = 107+/-61 ng/g). The sum of the three specific congeners (Parlar 26, 50 and 62) was within 10-55 ng/g. The ratio of the sum of the three congeners to the total toxaphene varied between 8% and 25% in the fish samples but the ratio may be species specific. Our results suggest that consumption of these Yukon fish should have minimal risk of toxaphene exposure. PMID- 10819222 TI - Toxaphene levels in salmon (Salmo salar) from the Baltic Sea. AB - Three toxaphene congeners have been determined in salmon from the Swedish coastal environment using both supercritical fluid (SFE) and the traditional liquid/liquid extractions. The levels obtained using a modifier-free SFE technique, followed by group separation on a silica gel column, were by far much lower than concentrations obtained by SFE with a modifier or liquid/liquid extraction. The mean concentrations on fresh weight basis using a liquid/liquid extraction technique were 5.87, 8.70 and 1.59 microg/kg for CHBs 26, 50 and 62, respectively. There was a plausible relationship between the various fishing sites and the toxaphene levels. PMID- 10819223 TI - Determination of toxaphenes in fish and marine mammals. AB - An analytical method for the determination of toxaphene in biological materials using gas chromatography with an electron capture detector (GC-ECD) has been established and validated for three single congeners (chlorinated bornanes (CHB) 26, 50 and 62). The analytical method was based on a method for determination of PCB, DDT and other chlorinated pesticides. To include toxaphene congeners an extra step, adsorption chromatography on silica columns, was introduced to separate the pesticides from PCB. The recovery of CHB-26, 50 and 62 were 97+/ 11%, 94+/-10% and 99+/-12%, respectively. Samples from cod, ringed seal and polar bear from the Norwegian arctic environment have been analysed. The levels of CHB 26 and 50 found were 13-55 ng/g fat in cod, 1.3-7.7 ng/g fat in ringed seal and 0.4-119 ng/g fat in polar bear. The levels of CHB-62 were 2.0-13, 0.8-3.4, 0.2-11 ng/g fat in cod, ringed seal and polar bear, respectively. PMID- 10819224 TI - Congener-specific analysis of toxaphene in eggs of seabirds from Germany by HRGC NCI-MS using a carborane-siloxane copolymer phase (HT-8). AB - A method for the congener-specific analysis of toxaphene in eggs of seabirds from a monitoring program from the northern part of Germany was carried out. The method was optimized in most steps of the procedure: injection temperature, HRGC with an HT-8 column, ion source temperature and the MS detection mode NCI/SIM measuring the isotope clusters of [M] , [M-Cl]-, [M-HCl]- and [M-2Cl]-. The suitability of 1,4-exo, 7,8,9,10,10-heptachloro-5-methoxytricyclo [5,2,1,0(2,6)]dec-3,8-diene as internal standard was demonstrated. 14 toxaphenes with Parlar numbers and more than 60 unknown toxaphenes could be identified. Spatial and temporal trends of toxaphene contamination are presented by using the Parlar 22 components standard for quantification. PMID- 10819225 TI - Priority assessment of toxic substances in life cycle assessment. Part I: calculation of toxicity potentials for 181 substances with the nested multi-media fate, exposure and effects model USES-LCA. AB - Toxicity potentials are standard values used in life cycle assessment (LCA) to enable a comparison of toxic impacts between substances. In most cases, toxicity potentials are calculated with multi-media fate models. Until now, unrealistic system settings were used for these calculations. The present paper outlines an improved model to calculate toxicity potentials: the global nested multi-media fate, exposure and effects model USES-LCA. It is based on the Uniform System for the Evaluation of Substances 2.0 (USES 2.0). USES-LCA was used to calculate for 181 substances toxicity potentials for the six impact categories freshwater aquatic ecotoxicity, marine aquatic ecotoxicity, freshwater sediment ecotoxicity, marine sediment ecotoxicity, terrestrial ecotoxicity and human toxicity, after initial emission to the compartments air, freshwater, seawater, industrial soil and agricultural soil, respectively. Differences of several orders of magnitude were found between the new toxicity potentials and those calculated previously. PMID- 10819226 TI - Priority assessment of toxic substances in life cycle assessment. Part II: assessing parameter uncertainty and human variability in the calculation of toxicity potentials. AB - Toxicity potentials are standard values used in life cycle assessment (LCA) to enable a comparison of toxic impacts between substances. This paper presents the results of an uncertainty assessment of toxicity potentials that were calculated with the global nested multi-media fate, exposure and effects model USES-LCA. The variance in toxicity potentials resulting from input parameter uncertainties and human variability was quantified by means of Monte Carlo analysis with Latin Hypercube sampling (LHS). For Atrazine, 2,3,7,8-TCDD and Lead, variation, expressed by the ratio of the 97.5%-ile and the 2.5%-ile, ranges from about 1.5 to 6 orders of magnitude. The major part of this variation originates from a limited set of substance-specific input parameters, i.e. parameters that describe transport mechanisms, substance degradation, indirect exposure routes and no effect concentrations. Considerable correlations were found between the toxicity potentials of one substance, in particular within one impact category. The uncertainties and correlations reported in the present study may have a significant impact on the outcome of LCA case studies. PMID- 10819227 TI - Electron microscopy study of biosorbents from marine macro alga Durvillaea potatorum. AB - Biosorbents derived from the biomass of marine algae have shown to have high uptake capacities for heavy metals and the internal structure has been generally assumed to be pseudo-homogenous. In this paper, the microstructures of biosorbents derived from Australian marine alga Durvillaea poratorum were analysed using scanning electron microscopy. The structural components of the biosorbent resembled fiber-like cylinders. The internal structure was a highly connected network of cylinders with varying sizes. Methods of drying and pre treatment of the biomass also affected the details of the internal structure. Calcium chloride followed by thermal treatment provided the most uniform cylinder networks for the biosorbents. Heavy metal Cu2+ and Cd2+ binding in the biomass was confirmed by using an electron probe microanalyser. PMID- 10819228 TI - The influence on partial order ranking from input parameter uncertainty. Definition of a robustness parameter. AB - The method of partial order ranking has been used within the environmental area for a variety of purposes as an attractive way of handling complex information. However, the environmental data are often associated with a significant degree of uncertainty. In this investigation the general nature of the influence from data uncertainty on the partial order ranking is analyzed. A Monte Carlo type analysis is performed in which a series of randomly formed data are used to test the influence of data uncertainty. The partial order ranking is interpreted, where the results are transferred to a one-dimensional ranking scale taking into account that not all elements are ranked with the same certainty. A simple general robustness parameter (E) in form of the expected number of comparisons for each ranking element is defined and correlated to the uncertainty analysis results. A simple equation relates E to the number of elements and the number of parameters, respectively. The magnitude of the ranking uncertainty is shown to increase rapidly when the E value decreases below 4-5 comparisons per element. When the E value exceeds 5 the ranking uncertainty becomes nearly constant and independent on the actual E value. PMID- 10819229 TI - Passenger aircraft cabin air quality: trends, effects, societal costs, proposals. AB - As aircraft operators have sought to substantially reduce propulsion fuel cost by flying at higher altitudes, the energy cost of providing adequate outside air for ventilation has increased. This has lead to a significant decrease in the amount of outside air provided to the passenger cabin, partly compensated for by recirculation of filtered cabin air. The purpose of this review paper is to assemble the available measured air quality data and some calculated estimates of the air quality for aircraft passenger cabins to highlight the trend of the last 25 years. The influence of filter efficiencies on air quality, and a few medically documented and anecdotal cases of illness transmission aboard aircraft are discussed. Cost information has been collected from the perspective of both the airlines and passengers. Suggestions for air quality improvement are given which should help to result in a net, multistakeholder savings and improved passenger comfort. PMID- 10819230 TI - Precipitation of carbonates by Nesterenkonia halobia in liquid media. AB - We investigated the precipitation of carbonates by Nesterenkonia halobia in a liquid medium at different concentrations of salts and incubation times. N. halobia only produced crystals at salt concentrations of 2.5%, 7.5% and 15%. At 20% salt concentration no crystal formation was observed. Calcite, aragonite and dolomite were precipitated in different quantities, depending on the salinity of the medium and incubation time. Scanning and transmission electron microscopy, microanalysis and electron diffraction were all used to study in detail the morphology, composition and internal structure of the bioliths. We propose a mechanism for biolith formation involving both biological and inorganic processes. PMID- 10819231 TI - Plant protection products: assessing the risk for terrestrial plants. AB - The fundamental data requirements for the authorization of plant protection products and the inclusion of active ingredients in Annex I of Council Directive 91/414/EEC (Council Directive of 15 July 1991 referring to placing plant protection products on the market (91/414/EEC). Official Journal of European Communities L 230, 19 August 1991) are described in the Annexes II and III of this Directive. Definite instructions with regard to preconditions for implementation and methodology (guidelines) concerning investigations with terrestrial plants are deficient. In addition to that, the uniform principles for the registration of plant protection products in the Member States described in Annex VI of the directive do not include any criteria concerning the risk assessment for non-target plants. However, plant protection products often show effects on non-target plants which need to be assessed as a requirement for the authorisation of the product. Hence, the German Federal Environmental Agency has developed a tiered approach to assess the effects of plant protection products on non-terrestrial plants. The risk is assessed using the effect-concentration evaluated in ecotoxicological tests and the environmental concentration predicted by validated exposure models. To protect non-target plants in terrestrial ecosystems assessment factors need to be considered. In the future, the risk for terrestrial plants needs to be addressed, also with regard to the revision of the Annexes of Directive 91/414/EEC. PMID- 10819232 TI - Autonomic regulation of islet hormone secretion--implications for health and disease. AB - The pancreatic islets are richly innervated by parasympathetic, sympathetic and sensory nerves. Several different neurotransmitters are stored within the terminals of these nerves, both the classical neurotransmitters, acetylcholine and noradrenaline, and several neuropeptides. The neuropeptides, vasoactive intestinal polypeptide, pituitary adenlyate cyclase activating polypeptide and gastrin releasing peptide are constituents of the parasympathetic nerves, whereas the neuropeptides galanin and neuropeptide Y are localised to sympathetic nerve terminals. Furthermore, the neuropeptide calcitonin gene-related peptide is localised to sensory nerves and cholecystokinin is also an islet neuropeptide, although the nature of the cholecystokinin nerves is not established. Stimulation of the autonomic nerves and treatment with neurotransmitters affect islet hormone secretion. Thus, insulin secretion is stimulated by parasympathetic nerves or their neurotransmitters and inhibited by sympathetic nerves or their neurotransmitters. The islet autonomic nerves seem to be of physiological importance in mediating the cephalic phase of insulin secretion, in synchronising the islets to function as a unit allowing oscillations of islet hormone secretion, and in optimising islet hormone secretion during metabolic stress, e.g. hypoglycaemia and neuroglycopenia. The autonomic nerves could also be involved in the islet adaptation to insulin resistance with possible implication for the development of glucose intolerance and Type II (non-insulin-dependent) diabetes mellitus. It is concluded that islet innervation, through the contribution of all branches of the autonomic nerves and several different neurotransmitters is of importance both for the physiology and pathophysiology of the islets. PMID- 10819233 TI - Erythromycin improves glycaemic control in patients with Type II diabetes mellitus. AB - AIMS/HYPOTHESIS: Erythromycin mimics the effect of the gastrointestinal hormone motilin by binding to its receptor and acting as a motilin agonist. We recently found that motilin stimulates insulin secretion at lower doses than doses required to stimulate gastric contractile activity. We studied the effects of erythromycin on insulin secretion and glycaemic control in patients with diabetes mellitus. METHODS: Inpatients (n = 34) with Type II (non-insulin-dependent) diabetes mellitus were randomly assigned to receive either erythromycin (400 mg orally three times a day, n = 19) or a placebo (n = 15) for 1 week (first study). Another 34 outpatients with Type II diabetes were also treated with erythromycin (200 mg orally three times a day, n = 17) or a placebo (n = 17) for 4 weeks (second study). Finally, nine inpatients with Type II diabetes and eight normal control subjects received intravenous erythromycin (10 mg x kg(-1) x h(-1)) or saline infusion and insulin secretion was examined (third study). RESULTS: Erythromycin lowered fasting blood glucose and fructosamine concentrations (p < 0.01) and increased basal as well as glucose-stimulated insulin secretion (p <0.05-0.01) (first study). Low doses of erythromycin treatment for 4 weeks also significantly improved glycaemic control in Type II diabetic patients (second study). Erythromycin infusion significantly increased plasma insulin and decreased glucose concentrations in Type II diabetic and control subjects and greatly potentiated glucose-induced insulin secretion in the latter (third study). CONCLUSION/INTERPRETATION: These results indicate that erythromycin given orally has an antidiabetogenic effect and therefore erythromycin derivatives that lack the antibacterial activity could have a therapeutic value in Type II diabetic patients. PMID- 10819234 TI - Enhanced escape of non-esterified fatty acids from tissue uptake: its role in impaired insulin-induced lowering of total rate of appearance in obesity and Type II diabetes mellitus. AB - AIMS/HYPOTHESIS: To estimate non-esterified fatty acids kinetics in patients with Type II (non-insulin-dependent) diabetes mellitus and obese subjects in the postabsorptive state and during hyperinsulinaemia using non-equilibrium tracer conditions. METHODS: We evaluated the effect of hyperinsulinaemia [euglycaemic clamp with insulin infused at 30 mU x kg(-1) x h(-1) (3-4 h) and 150 mU x kg(-1) x h(-1) (3 h)] on non-esterified fatty acid kinetics, traced with [14C]-palmitate using non-equilibrium tracer conditions in non-obese and obese healthy subjects and Type II diabetic patients (10 per group). Michaelis-Menten kinetics were applied for total non-esterified fatty acid disposal, which was assumed to be composed of total arterial plasma non-esterified fatty acid rate of appearance (equalling the rate of disappearance) and tissue uptake of non-esterified fatty acids derived from intravascular triglyceride hydrolysis. A model was developed to calculate the rate of escape of non-esterified fatty acids from tissue uptake and the net rate of tissue lipolysis. RESULTS: Total arterial plasma non esterified fatty acid rate of appearance was lower in non-obese healthy subjects than in the other groups at low insulin infusion (p < 0.05) and in obese Type II diabetic patients at high insulin infusion (p < 0.05). Plasma triglycerides were also lowest in non-obese healthy subjects during hyperinsulinaemia (p <0.05 from other groups). The rate of escape from tissue uptake decreased during hyperinsulinaemia (p < 0.05 for each group) but remained higher in obese Type II diabetic patients (p < 0.05 from non-obese healthy subjects). In contrast, net rate of tissue lipolysis was not different between the groups at baseline and its decline during hyperinsulinaemia (p < 0.05 for each group) was similar in all groups. CONCLUSION/INTERPRETATION: This study challenges the view that the antilipolytic effect of insulin is impaired in Type II diabetes and obesity. We suggest that a high plasma triglyceride concentration causes a higher escape of non-esterified fatty acids from tissue uptake, leading to an impaired suppression of total arterial plasma rate of appearance during a low degree of hyperinsulinaemia in obese subjects and Type II diabetic patients and during a high degree of hyperinsulinaemia in obese Type II diabetic patients. PMID- 10819235 TI - Mechanism of impaired nitric oxide synthase activity in skeletal muscle of streptozotocin-induced diabetic rats. AB - AIMS/HYPOTHESIS: The aims of our study were to investigate whether nitric oxide synthase (NOS) activity is impaired in skeletal muscle of insulin-deficient [Type I (insulin-dependent)] diabetic rats and if the case, to determine the mechanism of NOS dysregulation in this disorder. METHODS: Rats were rendered diabetic by streptozotocin injection (65 mg/kg, i.v.) and NOS activity and expression in gastrocnemius muscles were studied 1, 2, 3 or 4 weeks after diabetes induction. RESULTS: The diabetic state was associated with a progressive reduction (down to 42 % of control values after 4 weeks) in muscle NOS activity compared with control rats. Using reverse transcriptase-polymerase chain reaction, we could not detect statistically significant changes in the expression of either neuronal NOS (nNOS) or endothelial NOS (eNOS) mRNAs in diabetic muscle. The contents of nNOS and eNOS protein were, however, progressively reduced in muscle homogenates of diabetic rats and these alterations were prevented by insulin treatment. Subcellular fractionation of skeletal muscle showed that both nNOS and eNOS proteins are mainly localised to the plasma membrane with lower abundance in T tubules and not detectable in sarcoplasmic reticulum-enriched fractions. After 1 week of diabetes, eNOS protein content was decreased only in the plasma membrane whereas nNOS protein abundance was not affected at this time. Neither the expression nor the interaction of caveolin-1 and caveolin-3 with NOS enzymes was found to be altered in muscle of diabetic rats. CONCLUSION/INTERPRETATION: These results show that skeletal muscle NOS activity is impaired during the progression of insulin-deficient diabetes and reduced NOS activity is associated with a decreased abundance of both nNOS and eNOS proteins, which appears to involve post transcriptional mechanisms. PMID- 10819236 TI - Free and protein bound leptin are distinct and independently controlled factors in energy regulation. AB - AIMS/HYPOTHESIS: Leptin exerts important regulating effects on energy homeostasis and could have a central role in our understanding of obesity, diabetes mellitus and the metabolic syndrome. Leptin circulates in a free and protein bound form. The aim of the present study was to test whether both fractions of the leptin system can be selectively regulated and thus serve independent physiological roles. METHODS: Using specific radioimmunoassays we measured both leptin components in relation to BMI in healthy subjects before and after weight reduction and in hyperthyroid patients during correction of thyrotoxicosis. In the latter group body composition and resting energy expenditure was monitored. In addition, we measured serum and cerebrospinal fluid concentrations of free and bound leptin in patients with neurological disorders. RESULTS: Under all conditions free leptin concentrations reflected body fat mass. Bound leptin concentrations decreased during weight reduction but also after treatment of hyperthyroidism despite an increase in fat mass. Direct measurement of resting energy expenditure and bound leptin in hyperthyroid patients and under thyrostatic treatment showed a significant positive correlation of both variables. In contrast to free leptin whose transport into the cerebrospinal fluid appears to be saturated at low physiological concentrations of serum free leptin, bound leptin concentrations in the cerebrospinal fluid increased in parallel to serum concentrations over the whole physiologically relevant range. CONCLUSION/INTERPRETATION: Our data indicate a distinct role of free and bound leptin in the feedback regulating energy intake and expenditure and could have important implications for our understanding of the physiology and pathophysiology of leptin-dependent signalling. PMID- 10819237 TI - Serine residues 994 and 1023/25 are important for insulin receptor kinase inhibition by protein kinase C isoforms beta2 and theta. AB - AIMS/HYPOTHESIS: Inhibition of the signalling function of the human insulin receptor (HIR) is one of the principle mechanisms which induce cellular insulin resistance. It is speculated that serine residues in the insulin receptor beta subunit are involved in receptor inhibition either as inhibitory phosphorylation sites or as part of receptor domains which bind inhibitory proteins or tyrosine phosphatases. As reported earlier we prepared 16 serine to alanine point mutations of the HIR and found that serine to alanine mutants HIR-994 and HIR 1023/25 showed increased tyrosine autophosphorylation when expressed in human embryonic kidney (HEK) 293 cells. In this study we examined whether these mutant receptors have a different susceptibility to inhibition by serine kinases or an altered tyrosine kinase activity. METHODS: Tyrosine kinase assay and transfection studies. RESULTS: In an in vitro kinase assay using IRS-1 as a substrate we could detect a higher intrinsic tyrosine kinase activity of both receptor constructs. Additionally, a higher capacity to phosphorylate the adapter protein Shc in intact cells was seen. To test the inhibition by serine kinases, the receptor constructs were expressed in HEK 293 cells together with IRS-1 and protein kinase C isoforms beta2 and theta. Phorbol ester stimulation of these cells reduced wild type receptor autophosphorylation to 58 % or 55 % of the insulin simulated state, respectively. This inhibitory effect was not observed with HIR-994 and HIR 1023/25, although all other tested HIR mutants showed similar inhibition induced by protein kinase C. CONCLUSION/INTERPRETATION: The data suggest that the HIR domain which contains the serine residues 994 and 1023/25 is important for the inhibitory effect of protein kinase C isoforms beta2 and theta on insulin receptor autophosphorylation. PMID- 10819238 TI - Genetic and immunological characteristics of Type I diabetes mellitus in an Indo Aryan population. AB - AIMS/HYPOTHESIS: Our aim was to characterise the genetic and immunological features associated with Type I (insulin-dependent) diabetes mellitus in a cohort of Indo-Aryan children resident in the United Kingdom. METHODS: Children with Type I diabetes (n = 53), unaffected first-degree relatives (n = 146) and unrelated healthy control children (n = 54) were typed for alleles of the HLA DRB1, HLA-DQA1 and HLA-DQB1 genes. Islet cell antibodies and antibodies to glutamic acid decarboxylase, protein tyrosine phosphatase-2 (IA-2ic) and insulin were measured in the diabetic and control children. RESULTS: The DRB1*03.DQA1*05.DQB1*02 haplotype was positively associated with the disease, occurring in 78% of diabetic children compared with 22.6 % of healthy children (p(c) < 2.4 x 10(-5)). In simplex families, this haplotype was transmitted more frequently to the diabetic children than to their unaffected siblings (p < 1 x 10(-4)). The DRB1*04.DQA1* 03.DQB1*0302 haplotype was also transmitted preferentially to the diabetic probands (p < 0.025) but was not associated with disease in the case control study. Islet-related autoantibodies were detected in 89.6 % of diabetic patients compared with 11.8 % of control children (p < 1 x 10( 6)). Although protein tyrosine phosphatase-2 autoantibodies were detected more frequently among DRB1*04-positive diabetic patients compared with patients lacking this allele, the overall frequency of these autoantibodies was lower than observed in Europid diabetic subjects. This could reflect the absence of a disease association with DRB1*04 in the Indo-Aryan cohort. CONCLUSION/INTERPRETATION: Type I diabetes in our Indo-Aryan cohort is similar to the disease observed in Anglo-Europeans but has important immunogenetic differences. The low frequency of protein tyrosine phosphatase-2 autoantibodies among the Indo-Aryan diabetic children could have important implications for the design of future strategies for disease prediction in this population. PMID- 10819239 TI - Stability of autoantibodies and their relation to genetic and metabolic markers of Type I diabetes in initially unaffected schoolchildren. AB - AIMS/HYPOTHESIS: To study temporal changes in positivity for autoantibodies associated with Type I (insulin-dependent) diabetes mellitus and the relations between these antibodies, HLA-DQB1-risk markers and first-phase insulin response (FPIR) in non-diabetic schoolchildren. METHODS: The stability of the antibody status over 2 years was assessed in 104 schoolchildren initially positive for islet cell antibodies (ICA) or antibodies to the 65,000 M(r) isoform of the glutamic acid decarboxylase (GADA) or both and in 104 antibody-negative control children matched for sex, age and place of residence. All children were also studied for their first-phase insulin response and HLA-DQB1 alleles on the second occasion. RESULTS: On the second occasion 3 of the 98 initially ICA-positive children, 3/13 of those positive for antibodies to the IA-2 protein (IA-2A), 1/17 GADA-positive and 2/7 of those positive for insulin autoantibodies (IAA) tested negative for these antibodies. Children with IA-2A, GADA, IAA and multiple (> or = 2) antibodies had significantly lower first-phase insulin responses than the control children. In contrast, these responses did not differ between subjects with and without specific HLA-DQB1-risk alleles or genotypes. Of the six subjects with a considerably reduced first-phase insulin response three had multiple antibodies on both occasions but none of them had a DQB1 genotype conferring increased diabetes risk. Two subjects progressed to Type I diabetes within 3.4 years of follow-up, both of them having multiple antibodies and a considerably reduced first-phase insulin response but neither of them having a DQB1-risk genotype. CONCLUSIONS/INTERPRETATION: Positivity for diabetes-associated autoantibodies is a relatively stable phenomenon in unaffected schoolchildren, although conversion to seronegativity can occur occasionally. Our observations also indicate that DQB1 alleles associated with decreased susceptibility to Type I diabetes do not protect from impaired beta-cell function or from progression to overt disease in initially unaffected schoolchildren. PMID- 10819240 TI - Cloning and quantitative determination of the human Ca2+/calmodulin-dependent protein kinase II (CaMK II) isoforms in human beta cells. AB - AIMS/HYPOTHESIS: The Ca2+/calmodulin-dependent protein kinase II (CaMK II) is highly expressed in pancreatic islets and associated with insulin secretion vesicles. The suppression of CaMK II disturbs insulin secretion and insulin gene expression. There are four isoforms of CaMK II, alpha to delta, that are expressed from different genes in mammals. Our aim was to identify the isoforms of CaMK II expressed in human beta cells by molecular cloning from a human insulinoma cDNA library and to assess its distribution in humans. METHODS: The previously unknown complete coding sequences of human CaMK IIbeta and the kinase domain of CaMK IIdelta were cloned from a human insulinoma cDNA library. Quantitative determination of CaMK II isoform mRNA was carried out in several tissues and beta cells purified by fluorescence activated cell sorting and compared to the housekeeping enzyme pyruvate dehydrogenase. RESULTS: We found CaMK IIbeta occurred in three splice variants and was highly expressed in endocrine tissues such as adrenals, pituitary and beta cells. Liver showed moderate expression but adipose tissue or lymphocytes had very low levels of CaMK IIbeta-mRNA. In human beta cells CaMK IIbeta and delta were expressed equally with pyruvate dehydrogenase whereas tenfold lower expression of CaMK IIgamma and no expression of CaMK IIalpha were found. CONCLUSION/INTERPRETATION: Although CaMK IIdelta is ubiquitously expressed, CaMK IIbeta shows preferential expression in neuroendocrine tissues. In comparison with the expression of a key regulatory enzyme in glucose oxidation, pyruvate dehydrogenase, two of the four CaM kinases investigated are expressed at equally high levels, which supports an important role in beta-cell physiology. These results provide the basis for exploring the pathophysiological relevance of CaMK IIbeta in human diabetes. PMID- 10819241 TI - Testicular Sertoli cells exert both protective and destructive effects on syngeneic islet grafts in non-obese diabetic mice. AB - AIMS/HYPOTHESIS: Testicular Sertoli cells protect allogeneic islet grafts from rejection after transplantation into animals with chemically induced diabetes. The aims of this study were to determine whether Sertoli cells can protect syngeneic islets from autoimmune destruction after transplantation into non-obese diabetic (NOD) mice and, if so, whether protection is due to Sertoli cell expression of Fas ligand (FasL), believed to be the mechanism that protects against allograft rejection. METHODS: We compared the survival of syngeneic islets transplanted under the renal capsule of nonobese diabetic mice, alone and together with purified Sertoli cells prepared from testes of newborn nonobese diabetic mice. Additionally, we examined the composition of the islet and Sertoli cell co-transplants by immunohistochemistry to determine whether islet graft survival correlated with Sertoli cell expression of Fas ligand. RESULTS: Sertoli cell doses of 1, 2 and 4 x 10(6) cells produced a dose-dependent prolongation of median islet graft survival from 11 days (islets alone) to 32 days (islets + 4 x 10(6) Sertoli cells); addition of 8 x 10(6) Sertoli cells to the islet grafts decreased, however, median survival to 8 days. Immunohistochemical analysis of the islet and Sertoli cell co-transplants showed a correlation between Fas ligand expression by Sertoli cells and graft infiltration by neutrophilic leucocytes, leading to islet beta-cell destruction and diabetes recurrence. CONCLUSION/INTERPRETATION: Sertoli cells exert opposing effects on survival of syngeneic islet grafts in nonobese diabetic mice: Fas ligand-dependent neutrophil infiltration and graft destruction, and Fas ligand-independent protection of the graft from autoimmune destruction. PMID- 10819242 TI - Response of Charcot's arthropathy to contact casting: assessment by quantitative techniques. AB - AIMS/HYPOTHESIS: This study used two different methods of quantitative bone scanning to study the relation between activity of Charcot's arthropathy and clinical variables over 12 months. METHODS: Skin temperature of affected and unaffected feet was measured at baseline and every 3 months for 12 months in 17 subjects. Eight subjects underwent a three-phase quantitative bone scan at baseline and 3 monthly for 12 months. Bone isotope uptake in a standard rectangular area over the foot and tibia was analysed by the bilateral scan method (the ratio of isotope uptake of affected and unaffected feet) and the unilateral scan method (the ratio of isotope uptake of affected foot and ipsilateral tibia). The affected foot was placed in a contact cast for an average of 8 months. RESULTS: At presentation the affected foot was hotter than the unaffected foot but the temperature became progressively cooler over 12 months. Median isotope uptake in the affected foot was 2.1% of the injected dose (interquartile range, IQR 1.9-3.0). In both scanning methods the ratio of uptake decreased after casting but at 12 months the affected foot still had more isotope uptake. There was a strong correlation between temperature difference and the ratio of uptake in the bilateral scan method (r = 0.90; p < 0.0001) but when using the unilateral scan method this relation was not significant (r = 0.1;p = 0.6). A strong relation existed between perfusion of the affected foot in the dynamic phase and isotope uptake in the delayed phase of the scans (r = 0.92; p < 0.0001). CONCLUSION/INTERPRETATION: Bone activity and skin temperature of Charcot's arthropathy can be measured quantitatively and both improve over 12 months with contact casting. There is a strong relation between perfusion and disease activity in this condition. PMID- 10819243 TI - Very low density lipoprotein subfractions in Type II diabetes mellitus: alterations in composition and susceptibility to oxidation. AB - AIMS/HYPOTHESIS: Type II (non-insulin-dependent) diabetes mellitus is associated with raised triglycerides and increased very low density lipoprotein cholesterol. The aim of this study was to assess if very low density lipoprotein subfraction composition and potential to oxidise were altered in this condition. METHODS: Very low density lipoprotein was separated into four subfractions (A-->D) by a novel, rapid ultracentrifugation procedure. Analysis of each subfraction included lipid and fatty acid composition. Preformed peroxides were measured spectrophotometrically and conjugated dienes were used as an indicator of in vitro lipid oxidation. RESULTS: In all results we compared patient and control subfractions. Mean fasting plasma glucose was 8.9 +/- 2.0 mmol/l in patients vs 5.1 +/- 0.4 mmol/l in control subjects (p < 0.001); patient HbA1c was 7.6 +/- 1.4%. Patient total lipid standardised for apo B was higher than controls in subfractions A, B and C; A, 201 vs 60; B, 191 vs 40; C, 63 vs 21; D, 29 vs 34 micromol lipid per mg apo B (p < 0.05). Preformed peroxides were higher in all patient subfractions compared with controls: A, 340 vs 48; B, 346 vs 42; C, 262 vs 28; D, 54 vs 16 nmol per mg apo B (p < 0.001). Patient subfractions A and D were more susceptible to in vitro oxidation. Monounsaturated fatty acids were lower in patients subfractions, 35.2 vs 36.7; B, 35.1 vs 38.7; C, 34.4 vs 36.5; D, 33.0 vs 35.5 as per cent total (p < 0.05). CONCLUSIONS/INTERPRETATION: These results indicate abnormalities in very low density lipoprotein subfraction composition and oxidation profile in Type II diabetic subjects, which are characteristic of more atherogenic particles and that may contribute to the development of cardiovascular disease in these patients. PMID- 10819244 TI - Intimal medial thickness of the carotid artery in South Indian diabetic and non diabetic subjects: the Chennai Urban Population Study (CUPS). AB - AIM/HYPOTHESIS: Increased intimal medial thickness (IMT) of the carotid arteries is considered a useful marker of atherosclerosis. The aim of this study was to compare the intimal medial thickness values in urban non-diabetic and diabetic South Indian subjects who have a high risk of coronary artery disease. METHODS: The subjects for this study were 140 diabetic and 103 non-diabetic control subjects matched with them for age and sex selected from The Chennai Urban Population Study which is an ongoing epidemiological study. Intimal medial thickness of the right common carotid artery was determined using high resolution B mode ultrasonography. RESULTS: The mean intimal medial thickness values of the diabetic subjects (0.95 +/- 0.31 mm) were significantly higher than those of the non-diabetic (0.74 +/- 0.14 mm) subjects (p < 0.001). Both in the normal and diabetic subjects, these values increased with age. At any given age, the diabetic subjects had higher values than the non-diabetic subjects but the difference reached statistical significance after age 50 years (p < 0.05). Intimal medial thickness showed a correlation with age, total cholesterol, LDL cholesterol, waist:hip ratio and systolic blood pressure in non-diabetic subjects and with age and duration of diabetes in the diabetic subjects. Multivariate linear regression analysis showed that age and diabetes were the major risk factors for intimal medial thickness. CONCLUSION/INTERPRETATION: Diabetic subjects have higher intimal medial thickness values than non-diabetic subjects. Diabetes and age are the most important risk factors associated with increased intimal medial thickness in this South Indian cohort. PMID- 10819245 TI - Learning and hippocampal synaptic plasticity in streptozotocin-diabetic rats: interaction of diabetes and ageing. AB - AIMS/HYPOTHESIS: Diabetes mellitus leads to functional and structural changes in the brain which appear to be most pronounced in the elderly. Because the pathogenesis of brain ageing and that of diabetic complications show close analogies, it is hypothesized that the effects of diabetes and ageing on the brain interact. Our study examined the effects of diabetes and ageing on learning and hippocampal synaptic plasticity in rats. METHODS: Young adult (5 months) and aged (2 years) rats were examined after 8 weeks of streptozotocin-diabetes. Learning was tested in a Morris water maze. Synaptic plasticity was tested ex vivo, in hippocampal slices, in response to trains of stimuli of different frequency (0.05 to 100 Hz). RESULTS: Statistically significant learning impairments were observed in young adult diabetic rats compared with controls. These impairments were even greater in aged diabetic animals. In hippocampal slices from young adult diabetic animals long-term potentiation induced by 100 Hz stimulation was impaired compared with controls (138 vs 218% of baseline). In contrast, long-term depression induced by 1 Hz stimulation was enhanced in slices from diabetic rats compared with controls (79 vs 92%). In non-diabetic aged rats synaptic responses were 149 and 93% of baseline in response to 100 and 1 Hz stimulation, compared with 106 and 75% in aged diabetic rats. CONCLUSION/INTERPRETATION: Both diabetes and ageing affect learning and hippocampal synaptic plasticity. The cumulative deficits in learning and synaptic plasticity in aged diabetic rats indicate that the effects of diabetes and ageing on the brain could interact. PMID- 10819246 TI - Association of MHC Class I chain-related A (MIC-A) gene polymorphism with Type I diabetes. AB - AIMS/HYPOTHESIS: A distinct family of MHC genes has been identified in the class III region and denominated MHC Class I chain-related genes (MIC). The MIC-A gene is located between the TNFA and the HLA-B genes. The aim of our study was to test the association of the polymorphism of the MIC-A gene with Type I (insulin dependent) diabetes mellitus and evaluate the interaction between MIC-A and TNFA, HLA-B, HLA-DR and HLA-DQ gene polymorphism. METHODS: Type I diabetic (n =95) and healthy (n = 98) Italian subjects were typed for exon 5 of MIC-A and for HLA DRB1, HLA-DQA1, HLA-DQB1 and TNFA alleles. All subjects were also typed for the presence of HLA-B8 or HLA-B15. RESULTS: The frequency of MIC-A5 was increased in diabetic subjects (53 % vs 15 %) (OR = 6.1) (corrected p, p(c) < 0.0005). Among HLA class II haplotypes, both HLA-DRB1*03-DQA1*0501-DQB1*0201 (DR3-DQ2) and DRB1*04-DQA1*0301-DQB1*0302 (DR4-DQ8) ("at-risk class II haplotypes") were positively associated with diabetes (OR = 6.7 and 6.0, respectively) (p(c) < 0.003). Also HLA-B8 was more frequent among Type I diabetic subjects than among healthy control subjects (OR = 2.8, p = 0.01). None of the TNFA alleles were statistically significantly associated with Type I diabetes. The MIC-A5 exon was negatively associated with age at clinical onset of diabetes (p = 0.012). Thus, 68 % diabetic subjects younger than 25 years and 29 % older than 25 years were carrying this allele. Both MIC-A5 and the at-risk class II haplotypes were independently associated with Type I diabetes and the combined association of the two markers had the highest relative risk (OR = 172). In subjects younger than 25 years, the OR of MIC-A5 was as high as 21.7 and was more than twofold that of at risk class II haplotypes (OR = 9.5). The MIC-A5 exon was not in linkage disequilibrium with any of the HLA-class I, class II or TNFA alleles studied. CONCLUSIONS/INTERPRETATION: The MIC-A gene polymorphism is associated with genetic risk for Type I diabetes and the combination of MIC-A5 and at-risk class II haplotypes is now to be seen as the strongest genetic marker for this disease. PMID- 10819247 TI - Reduced second phase insulin secretion in carriers of a sulphonylurea receptor gene variant associating with Type II diabetes mellitus. AB - AIMS/HYPOTHESIS: The sulphonylurea receptor is a subunit of the ATP-sensitive potassium channel in the pancreatic beta cell. Mutations at nt -3 of the splice acceptor site of exon 16 and a silent mutation in exon 18 of the gene for the sulphonylurea receptor (SUR1) associate with Type II (non-insulin-dependent) diabetes mellitus in several independent populations. We investigated whether these gene variants associate with changes in the pattern of glucose-stimulated insulin secretion. METHODS: Subjects who had normal glucose tolerance (n = 67) and subjects with an impaired glucose tolerance (n = 94), originating from two independent studies, were included in the study. Beta-cell function and insulin sensitivity were assessed by the hyperglycaemic clamp. RESULTS: Frequencies of the exon 16 -3t allele in the normal and impaired glucose tolerant groups were 46% and 44% respectively (p = NS). The more rare exon 18 T allele showed frequencies of 5 and 7% respectively (p = NS). We observed an approximately 25% reduced second-phase insulin secretion in carriers of the exon 16 -3t allele in both groups (p < 0.05). Estimates of insulin sensitivity did not show differences between carriers and non-carriers. The variant in exon 18 and the combined presence of variants in exon 16 and exon 18 were not associated with differences in insulin secretion or insulin sensitivity in our study groups. CONCLUSION/INTERPRETATION: The diabetes associated exon 16 -3t variant of the SUR1 gene associates with a functional change of the beta cell as reflected by reduced second-phase insulin secretion in response to a standardized hyperglycaemia in normal and impaired glucose tolerant subjects. PMID- 10819248 TI - R127W-HNF-4alpha is a loss of function mutation but not a rare polymorphism and causes Type II diabetes in a Japanese family with MODY1. AB - AIMS/HYPOTHESIS: Mutations in the hepatocyte nuclear factor (HNF)-4alpha gene cause the type 1 form of maturity-onset diabetes of the young (MODY1). The R127W mutation is a missense mutation located in the T-box region of HNF-4alpha that was first identified in a Japanese family with MODY. We have examined the functional properties of this mutation in order to clarify the molecular basis of MODY1. METHODS: The intracellular localisation, DNA binding ability, transactivation activity and functional synergism with the coactivator CREB binding protein (CBP) of R127W-HNF-4alpha were investigated. RESULTS: The nuclear import and functional synergy with CBP of R127W-HNF-4alpha were normal. The DNA binding ability of the mutant was decreased as was its transcriptional activation of the HNF-1alpha and L-type pyruvate kinase (PKL) genes. CONCLUSION/INTERPRETATION: The R127W mutation seems to be a loss-of-function mutation. PMID- 10819249 TI - The role of tumour necrosis factor (TNF)-alpha resistance in obesity and insulin resistance. PMID- 10819250 TI - Obligatory information on patient-oriented end point outcome. PMID- 10819251 TI - Randomized controlled trials remain fundamental to clinical decision making in Type II diabetes mellitus: a comment to the debate on randomized controlled trials (For debate) PMID- 10819252 TI - Informed consent in neurosurgery: patients' recall of preoperative discussion. AB - OBJECTIVE: Informed consent (IC) is an important principle of modern medicine and the quality of the process is likely to receive increasing attention in future due to complex surgical procedures and a development of social mistrust for medical treatment. Medico-legal action is also becoming an important influence on IC, in particular the extent of warning to be given about the degree of risk. Evaluation of IC, however, encounters various problems. One key element of a knowledgeable decision is an analysis based on the disclosed risks. METHODS: In a prospective study, 104 consecutive patients were asked to write down as many items of the list of risks as possible two hours after consent interview. The sample included adults (mean age: 52 years) without any significant neuropsychological impairment who underwent elective intracranial (52%) or spinal (48%) surgery. Consent interview based on the proposals of the Deutsche Gesellschaft fur Neurochirurgie and the Berufsverband Deutscher Neurochirurgen 1998. The average number of disclosed risks was 32 in intracranial and 25 in spinal surgery. According to the results in the literature 6 'typical major risks' of the proposed treatment were selected. RESULTS: The median value of the total of recalled risks was 4 in the spinal group and 5 in the intracranial group. The mean score of general information retention was 18%. 65% of the patients did not recall more than 2 of 6 'typical major risks'. No significant correlation between recall performance and age or education was found. CONCLUSION: The general principles of memory apply and have implications in states of emotional arousal. Whereas education and age, unlike in previous reports, did not appear to influence recall, thus indicating that we had succeeded in tailoring IC to the individual, recall rate was low in most cases. Physicians should highlight the discrepancy and conflict between the requirements for the defence of medical practice in the law courts and the actual interests of patients. They should promote research to establish what really is 'appropriately informed consent'. PMID- 10819253 TI - Hydrostatic and hydrodynamic considerations in shunted normal pressure hydrocephalus. AB - Symptoms of overdrainage, a frequent complication following shunting procedures especially in patients with normal-pressure hydrocephalus (NPH), are presumed to be caused by siphoning: gravity-enhanced flow of cerebro spinal fluid (CSF) through shunts in upright position. A critical reappraisal of siphoning and overdrainage (OD) is presented considering the occurrence of subdural hygromas (SDH) encountered in a group of patients shunted for NPH. The patients participated in the multicentre Dutch NPH study: 101 NPH patients were randomised for treatment with shunts with low or medium-high working pressure. Treatment with shunts with low working pressure resulted in an increased incidence of SDH (iSDH). Current physiological concepts of overdrainage are discussed and criticised. A new theory is presented on the basis of general physiological considerations. The theoretical model proposed yields a revised explanation for the origin of extremely low ventricular pressure in the upright position found with overdrainage. The efficacy of siphoning-preventing shunts and subtemporal decompression is explored in the light of the proposed theory. PMID- 10819254 TI - Magnetic resonance-guided stereotactic biopsies: results in 100 consecutive cases. AB - BACKGROUND: We present the results of 100 consecutive magnetic resonance (MR) guided biopsies in cases where computerised tomography (CT) guiding was considered dangerous or impossible. METHOD: MR guiding was preferred to CT guiding for cases where lesions were located in the central area, or were not clearly visible on CT scan, or where the visualization of vessels was considered necessary. For most of the patients, calculation of target co-ordinates was performed using dedicated software enabling trajectory previsualization. There were 62 cases of contrast enhanced lesions, 32 cases of lesions without contrast enhancement, and 6 cases of very small lesions appearing hyperintense on T2 weighted images. FINDINGS: Biopsies allowed a histological diagnosis in 92 cases. In 8 cases, the biopsy was negative (necrosis, gliosis or normal brain tissue). Three patients had a transient worsening of their neurological disturbances. Two patient had a non-regressive loss of motor function. No patient died. INTERPRETATION: MR guiding for stereotactic biopsies was effective for CT invisible or ill-defined lesions, lesions located in functional or densely vascularized areas and in the brain stem. The rate of postoperative complications was equivalent to or less than that reported in series of CT-guided biopsies. PMID- 10819255 TI - Usefulness and pitfalls of intraoperative spinal motor evoked potential recording by direct cortical electrical stimulation. AB - OBJECTIVE: The aim of this study is to investigate the usefulness and problems with spinal motor evoked potential (MEP) recording, especially the reasons for failed recording. We report our personal experience over the last 8 years in patients with lesions adjacent to the primary motor cortex. METHODS: MEP records of 50 consecutive patients were retrospectively reviewed. MEP was recorded by a catheter electrode inserted in the cervical epidural space. Stimulation electrodes were placed on the cortical surface during surgery. SEP recording was also performed in 29 of 50 patients. RESULTS: MEP was obtained in 40 cases, and SEP was recorded in all 29 cases. The central sulcus was identified in 93% of patients in whom both MEP and SEP were performed, whereas in only 86% of patients who underwent only MEP. The main reason for MEP failure were inadequate exposure of the motor cortex, pre-existing hemiparesis and technical errors. Postoperative deterioration of motor function was closely related to intra-operative MEP changes. CONCLUSION: MEP is a useful tool to determine the motor cortex and to predict postoperative motor function. However, precise pre-operative craniotomy planning and combination with intra-operative SEP is essential to reduce the MEP failure. PMID- 10819256 TI - Intra-operative mapping of the motor cortex during surgery in and around the motor cortex. AB - The intra-operative use of neurophysiological techniques allows reliable identification of the sensorimotor region, and constitutes a prerequisite for its anatomical and functional preservation. The present prospective study combines monopolar cortical stimulation (MCS) with the recording of phase reversal of somatosensory evoked potentials (SEP-PR) in a protocol for the intra-operative mapping of the motor cortex. Functional mapping of the motor cortex by SEP-PR and MCS was performed in 70 patients during surgery in and around the motor cortex. The central sulcus was identified by SEP-PR. Cortical motor mapping was then performed by monopolar anodal (400 Hz) stimulation. Motor responses were recorded by needle electrodes placed in the muscles of the contralateral extremities. Surgery was performed under general anaesthesia without muscle relaxants. Intra operative localization of the central sulcus by SEP-PR was possible in 68 patients (97.14%). Motor evoked potentials (MEP) were elicited following MCS in 67 cases (95.7%). In 3 cases no MEP was recorded, not even after maximal stimulation intensity, the central sulcus being localized by SEP-PR only. On the other hand, MCS allowed localizing the motor cortex in the 2 cases with no recordable SEP-PR. Thus, combining SEP-PR and MCS allowed intra-operative localization of the sensorimotor cortex in 100% of the cases. PMID- 10819257 TI - Neovascularization (angiogenesis) after revascularization in moyamoya disease. Which technique is most useful for moyamoya disease? AB - The effects of direct and indirect revascularization for moyamoya disease were analyzed for each donor artery to determine which surgical procedure is most useful for the induction of neovascularization. In the past 12 years, 85 patients with moyamoya disease were surgically treated by combined surgery consisting of indirect revascularization via encephalo-duro-arterio-myo-synangiosis (EDAMS) and direct revascularization via the superficial temporal artery and the middle cerebral artery (STA-MCA) bypass. Among those patients, the post-operative changes in digital subtraction angiography were examined in 56 sides, including 34 sides in paediatric cases and 22 sides in adult cases. The neovascularization after indirect revascularization using the 1) superficial temporal artery (skin), 2) middle meningeal artery (dura mater), 3) deep temporal artery (temporal muscle) was analyzed. As results, in paediatric cases, the deep temporal artery and middle meningeal artery induced good neovascularization. However, the induction of neovascularization from the superficial temporal artery was not always good in most pediatric and adult cases. On the other hand, the direct bypass was useful in 90% of adult cases. In indirect revascularization surgery for moyamoya disease, the temporal muscle (the deep temporal artery) and the dura mater (the middle meningeal artery) are useful donors to the ischemic brain. The simple encephalo-arterio-synangiosis is not always effective. The direct bypass is a useful technique for adult moyamoya disease. PMID- 10819258 TI - The angiogenesis marker ED-B+ fibronectin isoform in intracranial meningiomas. AB - Fibronectins (FNs), adhesive glycoproteins mainly expressed in the extracellular matrix, are polymorphic molecules whose various isoforms are dependent on alternative splicing patterns. The isoform containing the ED-B sequence and occurring in foetal and neoplastic tissues (oncofoetal or B+FN) has been previously recognized as a marker for angiogenesis. The distribution of this isoform was analyzed in a consecutive series of 134 surgically obtained intracranial meningiomas, using specific monoclonal antibodies. Oncofoetal FN was found to be widely distributed in the vessels of anaplastic meningiomas, with its expression being restricted in the vasculature of the typical subtypes. and absent in the neighbouring cerebral tissue. The ubiquitous vascular expression of B+FN in meningiomatous malignancies might provide a potential target for the in vivo delivery of angiosuppressive agents. PMID- 10819259 TI - Outcome in Cloward anterior fusion for degenerative cervical spinal disease. AB - The Cloward ventral interbody fusion is often employed for treatment of cervical degenerative disease. The present study was aimed at evaluating results and complications in this classical type of autologous bone graft procedure in a cohort of patients with radiculopathy (RP) or myeloradiculopathy (MRP). Indications for and limitations of the technique were investigated by retrospective data analysis in a series of 106 patients (30 females and 76 males). These underwent single or multiple level Cloward fusion in a total of 145 levels. Neuroradiological investigations included lateral and antero-posterior cervical spine X-rays, axial CT scans, and MRI. The presence of postoperative ossification and stable bony fusion in the fused segments was confirmed by X-rays and, when necessary, by CT. The median postoperative follow-up period was 6.5 years (range 4-10.5 years). Short term outcome in RP patients was good in 26 cases (92.9%) and fair in 2 cases (7.1%). A good short term outcome was seen in 55 MRP patients (70.5%), a fair outcome in 20 patients (25.6%), and a poor outcome in 3 patients (3.8%). Patients having myelopathy signs for less than 1 year had a significantly better outcome than those with clinical signs for more than 1 year (p < 0.05). MRP patients below the age of 40 years had a significantly better outcome than those above the age of 40 (p < 0.05). In the long term, radiculopathy was cured or significantly improved in 92.8% of cases, and myeloradiculopathy in 64%. One year after surgery, there were 139 stably fused segments (96%) and 6 segments showing osseous non-union (4%). Plain lateral radiographs demonstrated, besides the bony fusion in the respective segment, relatively frequent graft collapse with slight to severe correction losses and kyphotic deformity of the cervical spine. However, these findings did not necessarily correlate with the clinical outcome. Autologous bone graft harvesting caused a rather high short-term morbidity with donor site pain and/or wound haematoma in 33% of the cases. These surgery-related complications, however, were of a temporary nature, as long-term complications (cutaneous hypaesthesiae) were found in 2 patients (1.8%) only. In conclusion, Cloward anterior cervical fusion for degenerative spinal disease is a relatively simple and safe surgical procedure with favourable short and long term results. In our hands, graft donor site complications dominate the side effects of surgery, and the percentage of non-unions is rather low. Because of the relatively frequent bone graft collapse and the late loss of postural correction of the spine, we cannot recommend the Cloward type fusion for multisegmental procedures. In such cases, an instrumented plate fusion should be carried out in order to prevent graft collapse and non union, and to allow for a shorter convalescence period. PMID- 10819260 TI - Surgery for syringomyelia: an analysis based on 163 surgical cases. AB - OBJECT: The authors analyzed the cases of 163 patients with syringomyelia to assess the appropriate surgical procedure. METHODS: Depending on the aetiological factors and treatment considerations the series was classified into three groups. Group I were cases where there was no definite demonstrable aetiological factor; Group II cases had basilar invagination and/or Chiari malformation; and Group III consisted of cases where the syrinx was secondary to an obvious aetiology, such as a mass lesion either in the posterior cranial fossa or in the spine or a severe kyphotic spinal deformity. Post-traumatic syringomyelia and syrinx in association with spina bifida were not studied. CONCLUSIONS: We concluded that for Group I cases syringosubarachnoid shunting is the ideal form of treatment. In Group II cases foramen magnum bony decompression is satisfactory and physiological. Good results were obtained even in cases where either a foramen magnum decompression alone or in combination with a syringo-subarachnoid shunt was done. Only syringosubarachnoid shunt (without a foramen magnum decompression) in Group II cases was found to produce poor outcome. Group III cases should be treated for the primary aetiological problem. Only syrinx drainage procedure without treatment of aetiology in these cases produced poor results. It was observed that clinical outcome rather than radiological improvement is the reliable indicator of the surgical result. PMID- 10819261 TI - 290 surgical procedures for ulnar nerve entrapment at the elbow: physiopathology, clinical experience and results. AB - Ulnar nerve entrapment at the elbow is an important and relatively frequent pathological condition that may be related to different causes depending on individual or external factors. The cause of the nerve lesion is also idiopathic in about one-quarter to one-third of cases. This variable aetiopathogenetic presentation has often suggested different diagnostic and clinical approaches and, moreover, various surgical procedures. We present our 8-years surgical experience with 290 cases of ulnar nerve entrapment at the elbow analysing the salient clinical features and the results of the surgical treatment in the light of the relevant literature available on this topic. PMID- 10819262 TI - The relationship between cerebral blood flow velocities and the amount of blood clots in computed tomography after subarachnoid haemorrhage. AB - 127 patients with aneurysmal subarachnoid haemorrhage (SAH) were analyzed for the relationship between the amount of blood clots as detected by initial computed tomography (CT) up to 48 hours after SAH and changes of blood flow velocities as measured using transcranial Doppler ultrasonography (TCD). All patients were operated on within 72 hours after SAH. Patients who presented with remarkable brain oedema or with pathological intracranial pressure (ICP) due to mass effects of a haematoma, and who were in a poor neurological condition classified according to Hunt-Hess as grade V were excluded from this study. Serial TCD examination of the middle cerebral arteries (MCA) and anterior cerebral arteries (ACA) started within 48 hours after SAH and were performed daily up to three weeks. A statistically significant correlation between blood load designated according to Fisher's grading as group CT I-CT IV and mean flow velocities (MFV) was found in groups CT I, II, and III. High values of MFV in MCA examinations were noted in patients with severe SAH (group CT III)--161 cm/s, and low values in patients without SAH (group CT I)--119 cm/s. Patients with haematocephalus and/or haematoma without a mass effect (group CT IV) had lower blood flow velocities than patients with severe SAH (group CT III) but values were higher than in patients without SAH (group CT I). The number of days for which MFV in the MCA was > 120 cm/s and was statistically (p < 0.05) correlated with the amount of blood clots as observed in the respective CT (in group CT I, II, and III). MFV values in the anterior cerebral artery (ACA) were lower than those obtained in the middle cerebral artery (MCA) in all groups. Statistically significant (p < 0.05) differences were noted between groups CT I and CT III (first and third week) and between groups CT I and CT IV (third week). If the SAH was extensive in the CT scan, pathological values of MFV > 90 cm/s were observed in ACA, and this was more pronounced in group CT III than in group CT IV. Blood flow velocities obtained via TCD were registered to compare side-to-side differences and particularly high differences were observed in patients with severe SAH. It is concluded that the amount of blood clots in the initial computed tomography after SAH is significantly correlated with cerebral blood flow velocity measurements by TCD. PMID- 10819263 TI - Comparison of 3-D stereoscopic MR imaging with pre and post lesion recording in pallidotomy. AB - OBJECTIVES: A quantitative on-line analysis of electrical activity in the pallidum of Parkinsonian patients has been developed to determine the focal point of lesioning. Additional recordings are made after the lesioning, to assess residual neural activity. A 3-D volume stereoscopic image system is proposed to display the complex anatomy and to superimpose the electrophysiological data into this system. The purpose of this study is to understand the complex pathophysiology in real-time anatomic/image space and determine the location and effect of lesioning residual energy sites. MATERIAL AND METHODS: Thirty patients undergoing 41 pallidotomies are presented. Neuronal activity from the pallidum is recorded using a semi-microelectrode. Based on this activity, lesioning is performed. Post-lesion recordings are made to determine the necessity of additional lesioning. 3-D volume MR images are acquired pre and postoperatively and compared for accuracy of lesion sites. A 3-D stereoscopic image system has been developed to depict basal ganglia geometry in the last 5 patients. Electrophysiological data are superimposed on this image system to show the surgeon the virtual position of the electrode tip. RESULTS: A stereoscopic 3-D volume MR image system has been developed. This system more efficiently and accurately visualizes and records the coordinates of high neural activity in the pallidum and post lesion residual activity. Post-lesion power analysis was carried out in 30 patients. Additional lesions were indicated in 8 and different trajectories in 6. CONCLUSION: Real-time visualization of neural recording, both pre and post-lesioning during pallidotomy, facilitates the surgeon's understanding of the spatiotemporal relationships of pathophysiological properties within the globus pallidus. PMID- 10819264 TI - A new cortical electrode for neuronavigation-guided intraoperative neurophysiological monitoring: technical note. AB - Intraoperative neurophysiological mapping and monitoring of eloquent brain areas can be combined with image-guided localisation to enhance the safety and efficacy of surgical procedures in the motor cortex. We designed a new type of cortical electrode which can be repeatedly placed on the cortical surface and allows accurate and reproducible stimulation by means of a navigation pointer. The newly designed device consists of a monopolar electrode contact for direct cortical stimulation, housed in a holder which allows placement, easy removal, and precise repeated placement of a surgical navigation pointer. It can be used for navigation-guided, high-frequency anodal monopolar cortical stimulation (MCS) for the mapping of eloquent cortex, and for monitoring of motor pathways. While the cortex is stimulated, compound muscle action potentials (CMAP) are recorded from muscles of the contralateral extremities and are assessed both qualitatively and quantitatively. When the device is used in combination with intraoperative navigation, the stimulation sites may optionally be registered or displayed on the system monitor. This allows repeated pinpointing and obviates the need for strip or grid electrodes in the operative field; although such electrodes may be useful for continuous monitoring, they often are in the surgeon's way. In addition, the primary and supplementary motor cortex can be mapped by determining the location of the sites of stimulation on surface-projected images of the cerebral cortex. PMID- 10819265 TI - Leukocyte-endothelium-interaction in pial vessels following global, cerebral ischaemia. AB - BACKGROUND: Investigations have shown an increase of leukocyte-endothelium interaction in a variety of organs following an ischaemic insult. To elucidate the role of leukocyte-endothelium-interaction following global, cerebral ischaemia the present study was performed. METHODS: Global, cerebral ischaemia was induced for twenty minutes by four-vessel-occlusion (PULSINELLI). Leukocyte endothelium-interaction was studied in the cerebral microcirculation using a rat closed cranial window and intravital microscopy. Leukocytes were stained intravenously using rhodamine 6G. Diameters of pial vessels, leukocyte centreline velocity and number of rolling or adhering leukocytes were determined off-line up to 2 h following global cerebral ischaemia. To confirm these results immunohistochemistry of the brain was performed. FINDINGS: Four-vessel-occlusion induced an iso-electric EEG, venular stasis and minimal rest flow in arterioles. Reperfusion yielded a significant increase of the arteriolar (p < 0.001) and a smaller increase of the venular diameters (p < 0.01). Up to 2 h after ischaemia no significant increase of the number of rolling or adhering leukocytes was measured which was confirmed by immunohistochemistry. INTERPRETATION: In contrast to other studies, in particular regarding focal cerebral ischaemia, an increase of leukocyte-endothelium-interaction in rat brain following 20 min of global cerebral ischaemia was not observed despite histological evidence of ischaemic damage. Thus in our model leukocytes seem not to contribute to the brain damage following global ischaemia. PMID- 10819266 TI - Benign recurrence of a cerebellar pilocytic astrocytoma 45 years after gross total resection. AB - OBJECTIVE AND IMPORTANCE: Cerebellar pilocytic astrocytomas are common benign tumours of childhood and mostly easy to treat because of their location and conformation. Nevertheless, unpredictable recurrence as well as the lack of features to predict the rare malignant changes after first removal, still represent a challenging problem. CLINICAL PRESENTATION: A 54 years old man presented with a recurrence of a cerebellar pilocytic astrocytoma 45 years after complete removal. INTERVENTION: Gross total resection was performed. The pathological examination confirmed the same histological pattern as the first tumour. DISCUSSION: Current opinions about management and recurrence of pilocytic astrocytomas are discussed with a review of the literature. PMID- 10819267 TI - Cerebral aneurysm associated with an anomalous hyperplastic anterior choroidal artery. AB - An unruptured internal carotid artery (ICA) aneurysm arising at the origin of a hyperplastic anomalous AchoA was identified together with a second unruptured middle cerebral artery aneurysm during angiography performed to investigate a striatal and intraventricular haemorrhage in a 55-years-old woman. The anomalous hyperplastic AchoA supplied the left temporal and occipital lobes, and the aneurysm arose proximal to its origin. The patient underwent clipping of the aneurysms, and intra-operative observation revealed that several perforating branches arose directly from the ICA between the AchoA and the ICA bifurcation. PMID- 10819268 TI - Microvascular decompression for treating glossopharyngeal neuralgia complicated by sick sinus syndrome. PMID- 10819269 TI - Ocular ischaemia with hypoplasia of the internal carotid artery associated with neurofibromatosis type 1. PMID- 10819270 TI - Empty sella syndrome following pituitary apoplexy. PMID- 10819271 TI - Single Price/Case Price Purchasing in orthopaedic surgery: experience at the Lahey Clinic. AB - BACKGROUND: Hospital revenues for orthopaedic operations are not keeping pace with inflation or with rising hospital expenses. In an attempt to reduce the hospital cost of orthopaedic operations by reducing the cost of operating-room supplies, we developed a Single Price/Case Price Purchasing Program for implants used in total hip arthroplasty, total knee arthroplasty, and total shoulder arthroplasty as well as for arthroscopic shavers and burrs, interference screws, and bone-suture anchors. METHODS: The Lahey Clinic asked orthopaedic vendors to supply all instruments, implants, and disposable items related to these selected products for one single price per unit or case. For example, a single price for total hip arthroplasty implants included instruments, acetabular cups, acetabular liners, acetabular screws, femoral stems, femoral heads, and stem centralizers, if required. The hospital implemented the Single Price/ Case Price Purchasing Program with a competitive-bid request for proposal. Surgeons evaluated the responses to the bidding process, and they made final decisions on product selection. RESULTS: The Single Price/Case Price Purchasing Program at the Lahey Clinic was successful in reducing the cost of orthopaedic implants and supplies. In the present article, we could not disclose the specific prices that we agreed to pay our vendors. The specific cost reductions were 32 percent for hip implants with a change of vendor, 23 percent for knee implants without a change of vendor, 25 percent for shoulder implants with a change of vendor, 45 percent for arthroscopic shavers and burrs without a change of vendor, 45 percent for interference screws without a change of vendor, and 23 percent for bone-suture anchors without a change of vendor. CONCLUSIONS: The Single Price/Case Price Purchasing Program at the Lahey Clinic allowed the hospital to reduce its cost of orthopaedic operations by lowering the cost of operating-room supplies. This cost reduction is important in a health-care economy in which hospital revenues per unit of service or care are decreasing. PMID- 10819272 TI - Isolated subtalar arthrodesis. AB - BACKGROUND: The purposes of this retrospective study were to review the results of isolated subtalar arthrodesis in adults and to identify factors influencing the union rate. The hypotheses were that (1) the overall outcome is acceptable but is not as favorable as previously reported, (2) complication rates, especially the nonunion rate, are higher than previously reported, and (3) factors contributing to a less favorable union rate can be identified. METHODS: Between January 1988 and July 1995, 184 consecutive isolated subtalar arthrodeses were performed in 174 adults (115 men and fifty-nine women) whose average age was forty-three years (range, eighteen to seventy-nine years). Eighty patients (46 percent) were smokers. The indications for the procedure included posttraumatic arthritis after a fracture of the calcaneus (109 feet), a fracture of the talus (thirteen feet), or a subtalar dislocation (thirteen feet); primary subtalar arthritis (thirteen feet); failure of a previous subtalar arthrodesis (twenty eight feet); and residual congenital deformity (eight feet). Rigid internal fixation with one or two screws was used for all feet. Bone graft was used in 145 feet; the types of graft material included cancellous autograft (ninety-four feet), structural autograft (twenty-nine feet), cancellous allograft (seventeen feet), and structural allograft (five feet). Bone graft was not used in the remaining thirty-nine feet. RESULTS: Clinical and radiographic follow-up examinations were performed for 148 (80 percent) of the 184 feet at an average of fifty-one months (range, twenty-four to 130 months) postoperatively. The average ankle-hindfoot score according to the modified scale of the American Orthopaedic Foot and Ankle Society (maximum possible score, 94 points) improved from 24 points preoperatively to 70 points at follow-up. Thirty feet had clinical evidence of nonunion. The union rate was 84 percent (154 of 184) overall, 86 percent (134 of 156) after primary arthrodesis, and 71 percent (twenty of twenty eight) after revision arthrodesis. The union rate was 92 percent (ninety-three of 101 feet) for nonsmokers and 73 percent (sixty-one of eighty-three feet) for smokers (p < 0.05). Intraoperative inspection revealed that 42 percent (seventy eight) of the 184 feet had evidence of more than two millimeters of avascular bone at the subtalar joint; all thirty nonunions occurred in this group (p < 0.05). A nonunion occurred in three of the five feet that had been treated with structural allograft and in two of the six feet in which the subtalar arthrodesis had been performed adjacent to the site of a previous ankle arthrodesis. After elimination of the subgroups of feet in patients who smoked, those that had had a failure of a previous subtalar arthrodesis, those that had been treated with a structural graft, and those that had had the subtalar arthrodesis adjacent to the site of a previous ankle arthrodesis, the union rate improved to 96 percent (seventy-three of seventy-six). Complications other than nonunion included prominent hardware requiring screw removal (thirty-six of 184 feet; 20 percent), lateral impingement (fifteen of 148 feet; 10 percent), symptomatic valgus malalignment (five of 148 feet; 3 percent), symptomatic varus malalignment (four of 148 feet; 3 percent), and infection (five of 184 feet; 3 percent). CONCLUSIONS: To the best of our knowledge, the present study includes the largest reported series of isolated subtalar arthrodeses in adults. Our results suggest that the outcome following isolated subtalar arthrodesis is not as favorable as has been reported in previous studies. The rate of union was significantly diminished by smoking, the presence of more than two millimeters of avascular bone at the arthrodesis site, and the failure of a previous subtalar arthrodesis (p < 0.05 for all). Other factors that probably affect the union rate include the use of structural allograft and performance of the arthrodesis adjac PMID- 10819273 TI - A biomechanical analysis of gait during pregnancy. AB - BACKGROUND: There are many anatomical changes during pregnancy that could potentially lead to substantial alterations in gait. Gait deviations may contribute to a variety of musculoskeletal overuse conditions associated with pregnancy, such as low-back, hip, and calf pain. Because we are aware of little research on this topic, the purpose of this study was to objectively analyze gait during pregnancy. METHODS: Three-dimensional gait analysis was performed on fifteen women during the second half of the last trimester of pregnancy and again one year post partum. Selected kinematic and kinetic parameters for the pregnancy and one-year postpartum conditions were compared with use of paired t tests (95 percent significance level). RESULTS: Overall, gait kinematics were remarkably unchanged during pregnancy. No evidence of a so-called waddling gait during pregnancy was found. Maximum anterior pelvic tilt during gait increased a mean of 4 degrees during pregnancy, although individual subject-to-subject variation (range, an increase of 13 degrees to a decrease of 10 degrees) was observed. Significant increases in hip and ankle kinetic gait parameters, however, were observed during pregnancy (p < 0.05). CONCLUSIONS: Significant increases in kinetic gait parameters during pregnancy (p < 0.05) explain how gait motion remained relatively unchanged despite increases in body mass and width as well as changes in mass distribution about the trunk. This finding indicates that during pregnancy there may be an increased demand placed on hip abductor, hip extensor, and ankle plantar flexor muscles during walking. PMID- 10819274 TI - Displaced scaphoid fractures treated with open reduction and internal fixation with a cannulated screw. AB - BACKGROUND: This study was performed to determine if the accuracy of screw placement was improved with use of the Herbert-Whipple cannulated screw compared with use of the AO/ASIF cannulated screw and also to evaluate the functional results in patients with an acute displaced fracture of the waist of the scaphoid treated with open reduction and internal fixation with a cannulated screw. METHODS: We retrospectively reviewed the results for thirty-five patients in whom an acute displaced fracture of the waist of the scaphoid had been treated with internal fixation with use of a cannulated screw. The patients were divided into two groups; Group 1 consisted of nineteen patients managed with a 3.5-millimeter cannulated AO/ASIF screw from 1990 through 1997, and Group 2 consisted of sixteen patients managed with a Herbert-Whipple screw from 1993 through 1997. RESULTS: There were no clinical or radiographic differences between the two groups. The average time to union (and standard deviation), confirmed with tomography, was 4.2 +/- 1.2 months for Group 1 and 4.0 +/- 1.2 months for Group 2. Both screws significantly improved the alignment of the scaphoid and decreased carpal collapse (p < 0.01). Importantly, the use of either cannulated screw improved the height-to-length ratio and the lateral intrascaphoid angle, which were correlated with an increase in the range of motion of the wrist (r = 0.584 and 0.625). In addition, both screws allowed for accurate placement in the central portion of the proximal pole. Regardless of the type of screw used, the time to union increased with increasing age of the patient (r = 0.665) and with increasing initial displacement of the fracture (r = 0.541). Within both groups, the time to union was longer for the patients who smoked (p < 0.01). CONCLUSIONS: Within both groups, cannulated screw fixation maintained the corrected fracture alignment and promoted healing and return of function. Our study shows cannulated screws to be a safe and effective method of treatment. PMID- 10819275 TI - Functional and oncological outcome of acetabular reconstruction for the treatment of metastatic disease. AB - BACKGROUND: Metastatic disease of the acetabulum can be painful and disabling. Operative intervention is indicated for patients who fail to respond adequately to nonoperative treatment. We evaluated the functional and oncological outcome of acetabular reconstruction after curettage for the treatment of refractory symptomatic acetabular metastases. METHODS: Fifty-five patients with metastatic disease of the acetabulum were treated with operative acetabular reconstruction combined with a total hip replacement. The most common primary tumor was carcinoma of the breast (eighteen patients), followed by carcinoma of the kidney (seven patients) and carcinoma of the prostate (seven patients). Forty (73 percent) of the patients presented with multiple skeletal metastases, and eighteen (33 percent) had associated visceral metastases. Twenty-eight (51 percent) had severe pain requiring continuous use of narcotics, twenty-four (44 percent) had moderate pain requiring periodic use of narcotics, and the remaining three (5 percent) had mild pain requiring use of non-narcotic analgesics. Eighteen (33 percent) of the patients could not walk, twenty-three (42 percent) needed a walker or crutches, twelve (22 percent) used a single cane, and two (4 percent) walked without assistive devices. Intralesional curettage of the tumor was performed in all of the patients. Fifty-four of the hips were reconstructed with a protrusio cup and one, with a hemipelvis endoprosthesis. Large defects were reinforced with cement and pin or screw fixation (the modified Harrington technique), which allowed transmission of weight-bearing forces to the remaining intact pelvis. Thirty-six acetabular reconstructions were performed with antegrade pins or cannulated screws; fifteen, with long retrograde screws; and four, with cement. RESULTS: The median period of survival was nine months. Patients with visceral metastases had a median period of survival of three months compared with twelve months for patients without visceral metastases (p < 0.001). Patients with breast cancer presented later in the disease process (p < 0.004) and lived longer than did those with other carcinomas (p < 0.004). Forty-five patients were evaluated three months after reconstruction. Thirty-four (76 percent) of them had relief of pain as determined by decreased use of narcotics. Nine of the eighteen patients who could not walk preoperatively regained the ability to walk. Fourteen of the seventeen patients who originally were able to walk in the community retained that ability. Thirty-three patients were available for evaluation at six months. Twenty-five (76 percent) still had relief of pain, and nineteen (58 percent) were able to walk and function in the community. Overall, fourteen (25 percent) of the fifty-five patients had moderate local progression of the disease, and five of these patients had failure of the fixation. Fourteen early complications developed in twelve (22 percent) of the patients. One patient (2 percent) died perioperatively. CONCLUSIONS: Patients who have acetabular metastases that are refractory to radiation and chemotherapy have a short life expectancy. The early, gratifying results of reconstruction validate the role of operative treatment as a short-term palliative procedure. Protrusio acetabular cups presumably compensate for deficiencies of the medial wall, while cement and pin fixation can be used effectively to reconstruct large defects in the acetabular column and dome. The low rate of fixation failure supports the biomechanical principles of the reconstruction. Generally, the reconstructions are sufficiently durable to exceed the life expectancy of the patients. PMID- 10819276 TI - Restoration of prehension with the double free muscle technique following complete avulsion of the brachial plexus. Indications and long-term results. AB - BACKGROUND: Recent interest in reconstruction of the upper limb following brachial plexus injuries has focused on the restoration of prehension following complete avulsion of the brachial plexus. METHODS: Double free muscle transfer was performed in patients who had complete avulsion of the brachial plexus. After initial exploration of the brachial plexus and (if possible) repair of the fifth cervical nerve root, the first free muscle, used to restore elbow flexion and finger extension, is transferred and reinnervated by the spinal accessory nerve. The second free muscle, transferred to restore finger flexion, is reinnervated by the fifth and sixth intercostal nerves. The motor branch of the triceps brachii is reinnervated by the third and fourth intercostal nerves to restore elbow extension. Hand sensibility is restored by suturing of the sensory rami of the intercostal nerves to the median nerve or the ulnar nerve component of the medial cord. Secondary reconstructive procedures, such as arthrodesis of the carpometacarpal joint of the thumb, shoulder arthrodesis, and tenolysis of the transferred muscle and the distal tendons, may be required to improve the functional outcome. RESULTS: The early results were evaluated in thirty-two patients who had had reconstruction with use of the double free muscle procedure. Twenty-six of these patients were followed for at least twenty-four months (mean duration, thirty-nine months) after the second free muscle transfer, and they were assessed with regard to the long-term outcome as well. Satisfactory (excellent or good) elbow flexion was restored in twenty-five (96 percent) of the twenty-six patients and satisfactory prehension (more than 30 degrees of total active motion of the fingers), in seventeen (65 percent). Fourteen patients (54 percent) could position the hand in space, negating simultaneous flexion of the elbow, while moving the fingers at least 30 degrees and could use the reconstructed hand for activities requiring the use of two hands, such as holding a bottle while opening a cap and lifting a heavy object. The results were analyzed to identify factors affecting the outcome. CONCLUSIONS: The double free muscle procedure can provide reliable and useful prehensile function for patients with complete avulsion of the brachial plexus. PMID- 10819277 TI - Initial symptoms and clinical features in osteosarcoma and Ewing sarcoma. AB - BACKGROUND: The time between the initial symptoms of osteosarcoma and Ewing sarcoma and the correct diagnosis and treatment is long. Over the last two decades, the prognosis for patients with these diseases has dramatically improved due to a new chemotherapy regimen. As a consequence, a limb-sparing operation has become an alternative to amputation. The aim of this study was to establish the initial symptoms and physical signs of osteosarcoma and Ewing sarcoma from the records of the first medical visit and to identify early characteristics of the diseases to shorten the delay to diagnosis. METHODS: A group of patients with osteosarcoma or Ewing sarcoma was identified from the Swedish Cancer Register of patients thirty years old and younger. Records from the first medical visit due to symptoms related to the bone tumor were obtained for 102 patients with osteosarcoma and forty-seven patients with Ewing sarcoma. RESULTS: Pain related to strain was reported by eighty-seven (85 percent) of the patients with osteosarcoma and thirty (64 percent) of those with Ewing sarcoma, but only twenty one (21 percent) of the patients with osteosarcoma and nine (19 percent) of those with Ewing sarcoma reported pain at night. Forty-eight (47 percent) of the patients with osteosarcoma and twelve (26 percent) of those with Ewing sarcoma related the onset of symptoms to minor trauma occurring around the same time. A palpable mass was noted in forty (39 percent) of the patients with osteosarcoma and sixteen (34 percent) of those with Ewing sarcoma at the first visit, and in most cases the tumor diagnosis was suspected. There was a broad spectrum of misdiagnoses; the most common was tendinitis, which was the initial diagnosis in thirty-two (31 percent) of the patients with osteosarcoma and ten (21 percent) of those with Ewing sarcoma. The doctor's delay (the period from the first medical visit due to the symptoms to the correct diagnosis) was longer for Ewing sarcoma than for osteosarcoma (nineteen weeks and nine weeks, respectively; p < 0.0001). CONCLUSIONS: An initial symptom of both osteosarcoma and Ewing sarcoma was pain, which was intermittent and often related to strain but not frequently felt at night. A history of trauma was common, but the clinical course often diverged from what was expected from trauma. The clinical course of osteosarcoma and particularly of Ewing sarcoma was not steadily progressive but intermittent, which often misled the doctor into believing that the condition was temporary. The most important clinical feature was a palpable mass, which was noted in more than one-third of the patients at the first visit. This finding emphasizes that a thorough physical examination is absolutely necessary. PMID- 10819279 TI - Relationship of peak height velocity to other maturity indicators in idiopathic scoliosis in girls. AB - BACKGROUND: Our aim was to compare height velocity data, obtained from clinical height measurements, for girls who had idiopathic scoliosis with the data for adolescents who did not have scoliosis. We also compared the growth data with chronological age, menarchal age, and Risser sign in terms of their accuracy in the prediction of growth and progression of the scoliosis. METHODS: One hundred and twenty of 371 patients in a database of girls managed with a brace for the treatment of idiopathic scoliosis had sufficient height data for us to quantify their growth peak. Height velocity data was generated from standing-height measurements obtained, in a scoliosis clinic, with a minimum six-month interval between measurements, and the timing of peak height velocity was calculated. The age at menarche was recorded from the patients' records. The Risser sign and Cobb angle were determined by a single observer. Progression of the scoliosis was defined as an increase in the Cobb angle of at least 10 degrees, compared with the curve magnitude at the time of the initial evaluation, after a minimum of six months. Progression to a magnitude requiring surgery was defined as progression of at least 10 degrees to a magnitude of 45 degrees or more. RESULTS: The height velocity plot grouped by peak height velocity showed a high peak and a sharp decline with values similar to those in normal populations. Extrapolating from percentile charts, 90 percent of our patients ceased growing by 3.6 years after peak height velocity. The growth peak was blunted (averaged over too long a period such that the data for the period of most rapid growth was averaged in with that for a period of slower growth) when chronological age, menarchal age, and Risser sign were used to predict growth; this indicated that these maturity scales grouped the patients poorly in terms of growth. The primary curve was progressive in eighty-eight of the 120 patients. Sixty of these patients had a curve of more than 30 degrees at peak height velocity, and in fifty (83 percent) of the sixty the curve progressed to 45 degrees or more. The remaining twenty eight patients had a curve of 30 degrees or less at peak height velocity, with only one curve (4 percent) progressing to 45 degrees or more. Peak height velocity also grouped patients for maximal progression of the curve more accurately than did the other maturity scales, as most of the curves progressed maximally at peak height velocity. There was a wider spread of timing of maximal progression when chronological age, menarchal age, and Risser sign were used to predict progression. CONCLUSIONS: Height velocities generated from clinical height measurements for patients with idiopathic scoliosis document the growth peak and predict cessation of growth reliably. Knowing the timing of the growth peak provides valuable information on the likelihood of progression to a magnitude requiring spinal arthrodesis. PMID- 10819278 TI - Aprotinin decreases exposure to allogeneic blood during primary unilateral total hip replacement. AB - BACKGROUND: Aprotinin, a hemostatic agent, regulates fibrinolysis, modulates the intrinsic coagulation pathway, stabilizes platelet function, and exhibits anti inflammatory properties through inhibition of serine proteases, such as trypsin, plasmin, and kallikrein. Aprotinin has been used successfully for many years in cardiac operations, and there have been preliminary investigations of its use in hip replacement operations. The objectives of this multicenter, randomized, placebo-controlled, double-blind trial were to evaluate the efficacy and safety of aprotinin as a blood-sparing agent in patients undergoing an elective primary unilateral total hip replacement and to examine its effect on the prevalence of deep-vein thrombosis in this population. METHODS: Seventy-three patients received a placebo; seventy-six patients, a low dose of aprotinin (a load of 500,000 kallikrein inhibitor units [KIU]); seventy-five, a medium dose of aprotinin (a load of 1,000,000 KIU, with infusion of 250,000 KIU per hour); and seventy-seven patients, a high dose of aprotinin (a load of 2,000,000 KIU, with infusion of 500,000 KIU per hour). The end points for the determination of efficacy were transfusion requirements and blood loss. Patients received standard prophylaxis against deep-vein thrombosis and underwent compression ultrasonography with color Doppler imaging of the proximal and distal venous systems of both legs to evaluate for the presence of deep-vein thrombosis. RESULTS: Aprotinin reduced the percentages of patients who required any form of blood transfusion (47 percent of the patients managed with a placebo needed a transfusion compared with 28 percent of those managed with low-dose aprotinin [p = 0.02],27 percent of those managed with high-dose aprotinin [p = 0.008], and 40 percent of those managed with medium dose aprotinin [p = 0.5]). Only 6 percent (twelve) of the 212 patients treated with aprotinin required allogeneic blood compared with 15 percent (ten) of the sixty-eight patients treated with the placebo (p = 0.03). Aprotinin decreased the estimated intraoperative blood loss (p = 0.02 for the low-dose group, p = 0.04 for the medium-dose group, and p = 0.1 for the high-dose group), the measured postoperative drainage volume (p = 0.4 for the low-dose group, p = 0.006 for the medium-dose group, and p = 0.000 for the high-dose group), and the mean reduction in the hemoglobin level on the second postoperative day (thirty-four grams per liter for the placebo group, twenty-eight grams per liter for the low-dose group [p = 0.000], twenty-six grams per liter for the medium-dose group [p = 0.000], and twenty-three grams per liter for the high-dose group [p = 0.0001). The rate of deep-vein thrombosis was similar for all groups. CONCLUSIONS: We concluded that aprotinin is safe and effective for use as a hemostatic agent in primary unilateral total hip replacements. In patients who are at high risk of receiving allogeneic blood, use of aprotinin may be of particular clinical and economic benefit. PMID- 10819280 TI - Osteocutaneous radial forearm free flaps. The necessity of internal fixation of the donor-site defect to prevent pathological fracture. AB - BACKGROUND: Osteocutaneous radial forearm free flaps have fallen from favor due to pathological fractures of the radius. The purposes of this study were to propose a means to decrease the rate of pathological fracture by prophylactic fixation of the donor-site defect and to evaluate this technique biomechanically. METHODS: Two groups of ten matched pairs of fresh-frozen cadaveric radii were harvested. In Group 1, an eight-centimeter length of radius comprising 50 percent of the cross-sectional area of the bone was removed to simulate an osteocutaneous radial forearm donor-site defect. This defect was created in one member of each pair, with the other bone in the pair left intact. In Group 2, both members of the ten matched pairs of radii had identical defects created as previously described. However, one radius in each pair had a twelve-hole, 3.5-millimeter dynamic compression plate placed across the segmental defect. In each group, five matched pairs were tested to failure in torsion and five matched pairs were tested to failure in four-point bending. RESULTS: In Group 1, the intact radius was a mean of 5.7 times stronger in torsion and 4.2 times stronger in four-point bending than the radius with the segmental resection. In Group 2, the radius that was ostectomized and fixed with a plate was a mean of 4.0 times stronger in torsion and 2.7 times stronger in four-point bending than the ostectomized radius. CONCLUSIONS: Removal of an eight-centimeter segment from the radius dramatically decreased both torsion and bending strength. Application of a plate over the defect in the radius significantly restored the strength of the radius (p = 0.01). PMID- 10819281 TI - Cemented rotating-platform total knee replacement. A nine to twelve-year follow up study. AB - BACKGROUND: Although the LCS (low contact stress) rotating-platform mobile bearing knee replacement has been used extensively, there have been few intermediate or long-term clinical and radiographic follow-up studies evaluating the device. The purpose of this study was to report the nine to twelve-year results of a consecutive series of patients who had a primary total knee replacement performed with this device. METHODS: Between November 1985 and November 1988, the senior author (R. C. J.) performed 119 consecutive total knee arthroplasties in eighty-six patients with LCS rotating-platform femoral and tibial components and a Townley all-polyethylene dome patellar component. All components were fixed with cement. The average age of the patients at the time of the operation was seventy years (range, thirty-seven to eighty-eight years). Fifty-two patients (seventy-six knees) were female, and thirty-four patients (forty-three knees) were male. The patients were evaluated with clinical knee ratings and radiographic analysis nine to twelve years following the knee replacement. RESULTS: At the time of the nine to twelve-year follow-up, sixty four patients (eighty-six knees) were alive, eighteen patients (twenty-eight knees) had died, and four patients (five knees) had been lost to follow-up. Of the 114 knees in the eighty-two patients for whom the final outcome was known, none required a reoperation and none had a dislocation of the mobile-bearing prosthesis. For the forty-five patients (sixty-six knees) who returned for final clinical and radiographic follow-up examinations at nine to twelve years, the average clinical and functional Knee Society ratings were 30 points (range, 2 to 70 points) and 44 points (range, 0 to 80 points) preoperatively and 90 points (range, 63 to 102 points) and 75 points (range, 30 to 100 points) at the final follow-up evaluation. The average Hospital for Special Surgery knee rating was 57 points (range, 28 to 80 points) preoperatively and 84 points (range, 59 to 97 points) at the final follow-up evaluation. The average active range of knee flexion was from 0 degrees (range, 0 to 10 degrees) to 102 degrees (range, 15 to 120 degrees) at the final follow-up evaluation. Seven of the sixty-six knees were painful anteriorly. There was no periprosthetic osteolysis and no evidence of loosening on follow-up radiographs. CONCLUSIONS: After nine to twelve years of follow-up, the cemented LCS rotating-platform knee replacement was found to be performing well, with durable clinical and radiographic results. PMID- 10819282 TI - Supracondylar osteotomy of the femur with use of compression. Osteosynthesis with a malleable implant. AB - BACKGROUND: The goal of treatment of a valgus deformity of the knee that is secondary to osteoarthritis of the lateral compartment is to obtain axial correction of the malalignment of the extremity. Osteosynthesis of the osteotomized femur with use of internal fixation and a stiff implant has not been as successful as expected. We evaluated the accuracy and maintenance of correction and the stability of fixation with a malleable plate after a supracondylar osteotomy of the distal aspect of the femur that was performed to correct a valgus deformity of the knee. METHODS: We performed an incomplete oblique osteotomy of the distal aspect of the femur in nineteen patients (twenty one knees) and stabilized the osteotomy site with a malleable semitubular plate, which was bent to form an angled plate, and lag-screws. Postoperatively, the patients were immediately encouraged to walk, with partial weight-bearing on the affected extremity. The mean age of the patients was fifty-seven years (range, thirty-nine to seventy-one years), and the mean duration of follow-up was five years (range, two to twelve years). RESULTS: In seventeen knees, the osteosynthesis withstood the mechanical loading that occurred during the postoperative functional rehabilitation program. Prolonged use of crutches or immobilization, or both, was necessary after the operation in three knees. The osteosynthesis failed in one knee. The loss of correction in eighteen knees, after bone-healing, averaged 1.7 degrees (range, 0 to 4 degrees). CONCLUSIONS: Our method of achieving osteosynthesis is based on the concept that inherent endogenous stability mechanisms can be mobilized by circumferentially compressing the two cortical tubes with the cut ends congruently apposed to each other. We believe that our technique provides an alternative to osteosynthesis with use of a stiff implant such as a fixed-angle blade-plate device. PMID- 10819283 TI - The orthopaedic forum. Falling in love again. PMID- 10819284 TI - The orthopaedic forum. The doctor-patient partnership. PMID- 10819285 TI - The anatomy of the lateral femoral cutaneous nerve, with special reference to the harvesting of iliac bone graft. PMID- 10819286 TI - Ethics in practice. AB - H. K. is a ninety-two-year-old woman with Alzheimer's disease and mild hypertension. She resides at a nursing home, where she transfers from bed to chair with maximal assistance. She presents to our emergency department with a painful right hip. Physical examination demonstrates a confused, elderly patient with significant right hip pain and shortening and external rotation of the lower extremity. Radiographs demonstrate a displaced intertrochanteric hip fracture. The patient lacks the capacity for informed consent. Her family is contacted to obtain consent for insertion of a compression screw. The family refuses to give consent, stating that the patient is too old and the surgery is too dangerous. PMID- 10819287 TI - The prisoner's dilemma. PMID- 10819288 TI - Proximal femoral osteotomy. PMID- 10819289 TI - Metallosis and metal-on-metal bearings. PMID- 10819290 TI - Role of heparin in compartment syndrome. PMID- 10819291 TI - Ophthalmological numismatics. A look at the past... PMID- 10819292 TI - Arisostatins A and B, new members of tetrocarcin class of antibiotics from Micromonospora sp. TP-A0316. I. Taxonomy, fermentation, isolation and biological properties. AB - Arisostatins A and B, new members of tetrocarcin class of antibiotics were isolated from the culture broth of an actinomycete strain. The producing strain, TP-A0316, was identified as Micromonospora sp. Arisostatins were obtained from the culture fluid by solvent extraction and chromatographic purification. They showed antibiotic activity against Gram-positive bacteria and antitumor activity. PMID- 10819293 TI - Arisostatins A and B, new members of tetrocarcin class of antibiotics from Micromonospora sp. TP-A0316. II. Structure determination. AB - Structures of arisostatins A and B were determined by spectroscopic analyses. Arisostatins were found to be new analogs of tetrocarcin A and possess an iso butanoyldigitoxose unit instead of the acetyldigitoxose one. NMR analyses of arisostatins and tetrocarcin A led to the revision of the anomeric configurations in the tetrasaccharide moiety of tetrocarcin A. PMID- 10819294 TI - Activation of fibrinolysis by SMTP-7 and -8, novel staplabin analogs with a pseudosymmetric structure. AB - Two novel staplabin analogs, SMTP-7 and -8, have been isolated from cultures of Stachybotrys microspora IFO 30018. Spectroscopic analyses showed that the SMTP-7 molecule consisted of two identical staplabin core structures and ornithine which bridges the two partial structures. In the SMTP-8 molecule, the bridging unit was lysine. At concentrations of 80 approximately 150 microM, the two compounds caused 2- to 12-fold increase in urokinase-catalyzed plasminogen activation, fibrin binding of plasminogen, and urokinase- and plasminogen-mediated fibrinolysis. These activities of SMTP-7 and -8 were two to ten times higher than those of staplabin and previously isolated SMTPs, which exerted such effects at concentrations ranging from 150 to 800 microM. PMID- 10819296 TI - Biological activity of guanacastepene, a novel diterpenoid antibiotic produced by an unidentified fungus CR115. AB - Fermentation extracts of culture CR115, an unknown plant endophyte originally isolated from Costa Rica, were found to be active against antibiotic-resistant bacteria. The metabolite responsible for activity was identified as a novel diterpenoid antibiotic guanacastepene (mol. wt. 374.47 and mol. formula C22H30O5). Mechanistic studies done in an E. coli imp strain suggested membrane damage as the primary mode of bactericidal action. This compound also lysed human RBCs and caused leakage of intracellular potassium from E. coli imp. PMID- 10819295 TI - Chaetoatrosin A, a novel chitin synthase II inhibitor produced by Chaetomium atrobrunneum F449. AB - Chaetoatrosin A, a novel chitin synthase II inhibitor, was isolated from the culture broth of fungus F449, which was identified as Chaetomium atrobrunneum F449. Chaetoatrosin A was purified by solvent partition, silica gel, ODS, preparative TLC, and Sephadex LH-20 column chromatographies, consecutively. The structure of chaetoatrosin A was assigned as 1,8-dihydroxy-3(2-hydroxypropionyl) 6-methoxynaphthalene on the basis of various spectroscopic analyses including UV, IR, mass spectral, and NMR. Its molecular weight and formula were found to be 262 and C14H14O5, respectively. ,Chaetoatrosin A inhibited chitin synthase II by 50% at the concentration of 104 microg/ml in an enzyme assay system. This compound showed antifungal activities against Rhizoctonia solani, Pyricularia oryzae, Botrytis cinerea, Cryptococcus neoformans and Trichophyton mentagrophytes. PMID- 10819297 TI - Enhancement of fibrinolytic activity of vascular endothelial cells by chaetoglobosin A, crinipellin B, geodin and triticone B. AB - Four fungal metabolites, chaetoglobosin A (CGA), crinipellin B (CPB), geodin (GE) and triticone B (TTB), were found to enhance fibrinolytic activity of bovine aortic endothelial cells. Plasmin generation on the cells was elevated 2- to 4 fold when treated with these agents at a concentration of 3 approximately 100 microM. These effects were dependent on plasminogen and inhibited by anti urokinase antibody. The effect of CGA, but not of CPB, GE and TTB, was abolished by cycloheximide. In a cell-free system, plasmin and urokinase activities as well as urokinase-catalyzed plasminogen activation were not enhanced by these agents. CGA, but not others, induced the production of urokinase in endothelial cells. CPB and GE accelerated plasminogen activator inhibitor- 1 (PAI-1) inactivation, and TTB caused direct, reversible inhibition of PAI-1. Thus, induction of urokinase by CGA and inhibition of PAI-1 by CPB, GE and TTB may, at least partly, account for the elevation of fibrinolytic activity of endothelial cells. PMID- 10819298 TI - Structures of ADP-ribosylated rifampicin and its metabolite: intermediates of rifampicin-ribosylation by Mycobacterium smegmatis DSM43756. AB - 23-(O-ADP-Ribosyl)rifampicin [RIP-TAs (3, Na+ form), RIP-TAf (4, H+ form)] was obtained as an intermediate in the conversion process of rifampicin (1) to RIP-Mb (2) that is mediated by cell homogenates of Mycobacterium smegmatis DSM43756 or of Escherichia coli carrying a mycobacterial mono(ADP-ribosyl) transferase gene, in the presence of NADH. 23-[O-(5'-Phosphoribosyl)]rifampicin (5, RIP-TAp) was also obtained by the reaction of rifampicin with NADH in the presence of a homogenate of M. smegmatis. The structures of 3, 4, and 5 were determined by means of MS and NMR analyses. PMID- 10819299 TI - DNase I induced DNA degradation is inhibited by neomycin. AB - Preparations of antimicrobials from biotechnological sources containing nucleic acids may serve as vector for the dissemination of resistance genes. An essential prerequisite for the acquisition of a new resistance phenotype in a transformational scenario is the availability of physically intact DNA molecules capable of transforming competent microorganisms. DNA is thought to be an easy target for catabolic processes when present in the natural habitat of bacteria (e.g. gastrointestinal tract, soil) due to the overall presence of nucleolytic enzymes. Aminoglycoside antibiotics are known to display a strong affinity to nucleic acids rendering these compounds to be primary candidates for exerting DNA protective functions in the gastrointestinal tract when applied orally during antibiotic chemotherapy. Using a DNase I protection assay it could be demonstrated that neomycin B at a concentration of 2 mM completely inhibited degradation of plasmid DNA in vitro. No inhibition of degradation was observed with streptomycin and kanamycin and the non-aminoglycoside antibiotics oxytetracycline and ampicillin under identical assay conditions. Thus, neomycin preparations may be able to promote structural integrity of contaminating DNA fragments in DNase-rich environments. PMID- 10819300 TI - Structure-activity relationships in the series of eremomycin carboxamides. AB - A series of new carboxamides of the glycopeptide antibiotic eremomycin was synthesized and investigated in vitro. The goal of the study was the comparison of the influence of the substituents introduced onto the eremomycin skeleton on the activity of these compounds against vancomycin susceptible and resistant bacterial strains. Eremomycin amides derived from amines with small substituents (C0 approximately C4) demonstrated antibacterial activity against vancomycin susceptible strains similar to that of the parent antibiotic and were inactive against vancomycin resistant strains. The derivatives of alkylamines with linear lipophilic substituents (like C10H21) were active against VanA and VanB enterococci strains with the scope of activity similar to that of N'-decyl or 7d CH2NH-decyl eremomycins described earlier. Eremomycin amides of 5-methoxy- and 5 benzyloxytryptamine were active both against vancomycin susceptible and resistant strains. The introduction of a spacer (lysine or piperazine) between the decyl and antibiotic moieties did not seriously influence antibacterial properties of the compounds in comparison with the corresponding derivatives without a spacer. The most active carboxamides are of interest for secondary modifications of the antibiotic. PMID- 10819301 TI - UCS1025A, a novel antibiotic produced by Acremonium sp. PMID- 10819302 TI - New strobilurins O and P from a mushroom. PMID- 10819303 TI - Preparation of 13-epi-selamectin by biotransformation using a blocked mutant of Streptomyces avermitilis. PMID- 10819304 TI - Anticancer and some biological activities of thiazinotrienomycin B. PMID- 10819305 TI - Syntheses of oxanosine and carbocyclic oxanosine derivatives as anti-HIV agents. PMID- 10819306 TI - A novel 1beta-methylcarbapenem, J-111,225: effects of the C-3 and C-5 stereochemistry of the pyrrolidinylthio side chain on antibacterial activities. PMID- 10819307 TI - Defining wild-type life span in Caenorhabditis elegans. AB - The nematode Caenorhabditis elegans reproduces predominantly as a self fertilizing hermaphrodite, and this drives laboratory populations to be homozygous at all genetic loci. Passaging of stocks can lead to fixation of spontaneous mutations, especially when the latter do not result in a selective disadvantage under laboratory conditions. Life span may be such a trait, since a comparison of six wild-type N2 lines derived from a common ancestor (but maintained separately in several laboratories) revealed four variants with median adult life spans ranging from 12.0 +/- 0.8 to 17.0 +/- 0.6 days at 20 degrees C. Fertility was also reduced in the two shortest-lived strains. We determined which life span most closely corresponds to that of the authentic wild type by two means. Firstly, N2 hermaphrodites were compared with seven C. elegans wild isolates. The latter were found to resemble only the longest-lived N2 strain. Comparison of male life spans of six lines also revealed additional strain variation. Secondly, life spans of F1 progeny issuing from crosses between N2 variants showed that short life spans were recessive, indicating that they result from loss-of-function mutations. We infer that the longest-lived N2 variant best resembles the original N2 isolate. This is the N2 male stock currently distributed by the Caenorhabditis Genetics Center. PMID- 10819308 TI - Increase of oxidatively modified protein is associated with a decrease of proteasome activity and content in aging epidermal cells. AB - For the process of aging in epidermal cells to be characterized, the status of oxidized and damaged protein accumulation and removal by the proteasome has been investigated. Modified protein content and proteasome activity were assayed in lysates of epidermal cells from donors of different ages. Increased levels of oxidized proteins, glycated proteins, and proteins modified by the lipid peroxidation product 4-hydroxy-2-nonenal were observed in cells from old donors. At the same time, a decline of chymotrypsin-like and peptidylglutamyl-peptide hydrolase activities of the proteasome was found in aging keratinocytes. This age related decline of the proteasome peptidase activities can be explained, at least in part, by a decreased proteasome content as observed by immunoblotting and enzyme-linked immunosorbent assay. In keratinocyte cultures, a decrease of proteasome activity and content was observed upon serial passaging. In cultures, as well as in skin, an inverse relationship was found between the aging marker 1 galactosidase and the proteasome content. These results suggest that proteasome is downregulated during replicative senescence as well as in aged cells in vivo, possibly resulting in the accumulation of modified proteins. PMID- 10819309 TI - Minireview: mechanisms by which the metabolic syndrome and diabetes impair memory. AB - Type 2 diabetes is associated with an increased risk of cognitive dysfunction. These effects seem particularly true for memory functions. This article examines how diabetes and the biological changes that occur with diabetes such as hyperglycemia, changes in insulin concentration, hypertension, and changes in lipid levels might lead to these alterations in cognitive functioning, with an emphasis on the mechanisms leading to changes in memory. PMID- 10819310 TI - Age-dependent changes in the midsized neurofilament subunit in sensory-motor systems of the cat brainstem: an immunocytochemical study. AB - This study documents age-related changes in the immunoreactivity of the medium molecular weight subunit of neurofilaments in sensory and motor neurons in the brainstem of the cat. In old age, there was a clear decrease in immunoreactivity in the following brainstem sensory and motor nuclei: sensory trigeminal, gracile, cuneate, and facial motor. Only a few neuronal perikarya and dendrites were labeled in these nuclei in old cats; moreover, when present, the labeling was weak. In contrast, in adult cats, these nuclei contained intensely stained neuronal perikarya and dendrites. In other sensory and motor nuclei of the brainstem, there was an obvious age-related increase in the immunoreactivity of the medium-molecular weight subunit of neurofilaments in the perikarya. Despite different patterns of age-related alterations in immunoreactivity within perikarya and dendrites in distinct brainstem regions, most sensory and motor axons in old cats were smaller than those in adult cats. A decrease in the medium molecular weight neurofilament subunit in the dendrites may be the basis for the dendritic atrophy that has been shown to occur in sensory nuclei in old animals. The decrease in axonal size is likely to be one of the causes of the decrease in axonal conduction velocity, in these neurons, that was reported in our previous studies. PMID- 10819311 TI - Delayed and blunted induction of mRNA for tissue plasminogen activator in the brain of old rats following pentylenetetrazole-induced seizure activity. AB - The ability of the rodent brain to support plasticity-related phenomena declines with increasing age. Here we investigated the extent to which old rats retain the capacity to initiate transcription for immediate early genes, particularly as it relates to brain plasticity, in response to a strong stimulus. The intraperitoneal administration of pentylenetetrazole (PTZ) to rats of various ages evoked tonic-clonic seizures. Using an RNA gel-blot and in situ hybridization analysis, we found that 1 hour after the onset of seizure, messenger RNA (mRNA) for tissue plasminogen activator (TPA) was increased approximately 3.7-fold in the hippocampi of 3-month-old rats. The levels of TPA mRNA in the hippocampi and cortices of 3-month-old rats returned to control levels by 3 hours after PTZ administration. The levels of TPA mRNA increased 2.5 fold in the hippocampi of 18-month-old rats and 1.8-fold in the brains of the 28 month-old-rats at 3 hours and returned to basal levels by 15 hours following PTZ treatment. Quantitatively similar increases were calculated for the cortex. At peak induction the transcripts were localized throughout the cortical layers of the 3-month-old rats, whereas the TPA mRNA expression was restricted to cortical layer V of the older rats. Our results suggest that although the aging brain retains the capacity to respond to chemically induced seizures, the induction of TPA mRNA is temporarily delayed and the levels are diminished with increasing age. Because TPA has been implicated in neuronal plasticity, this finding suggests that immediate early genes are important factors in the limited plasticity of the aging brain. PMID- 10819312 TI - Large neutral amino acid changes and delirium in febrile elderly medical patients. AB - A hypothesized but unexplored mechanism for delirium in older persons is that changes in plasma large neutral amino acid (LNAA) concentrations alter brain serotonin levels, result in neurotoxicity, or both. Therefore we performed a prospective study of 21 acutely febrile long-term-care residents to study the relationship between LNAA changes and delirium. Plasma LNAA concentrations were evaluated during illness and 1 month later. Delirium was diagnosed by using the Confusion Assessment Method. Other data included age, body mass index, cognitive impairment, comorbidity, gender, maximum temperature, and medication use. Seven subjects (33%) were delirious during febrile illness. Although the phenylalanine (PHE)/LNAA ratio was higher during illness in both delirious and nondelirious groups, a two-sample t test demonstrated that delirium was associated with a higher illness PHE/LNAA ratio (p = .03). The amount of change in PHE/LNAA from illness to recovery was not different between the delirious and nondelirious groups. Tryptophan/LNAA was not associated with delirium during illness or at recovery. These findings identify another potentially fruitful area of investigation for the prevention and treatment of delirium in older persons. PMID- 10819313 TI - Diabetes mellitus: a major disease of older persons. PMID- 10819314 TI - Epidemiology of hyperglycemia in elderly persons. AB - BACKGROUND: Our study used data collected in the Chung-Shing-Shin-Tseun community of Taiwan in May 1998 to evaluate the distribution of fasting glucose and the relation between hyperglycemia and the cardiovascular risk factors and sociodemographic factors in elderly persons. METHODS: Individuals aged 65 and over were recruited as study subjects. A total of 1,093 persons, out of 1,774 registered residents, were contacted in face-to-face interviews. The response rate was 61.6%. However, only 586 respondents took blood tests and completed questionnaires. Analysis in this study was based on these 586 subjects. The t test, chi-square analysis, and multivariate logistic regression were used to study the significant correlates of hyperglycemia. RESULTS: Of the individuals in our study, 66.0% were men and 34.0% were women. The mean age was 73.1 +/- 5.3 years. The mean values of fasting glucose were 5.5 +/- 1.6 mmol/L in elderly men and 5.7 +/- 2.1 mmol/L in elderly women. The hyperglycemic rates determined by modified World Health Organization criteria (> or =6.05 mmol/L) were 20.2% in elderly men and 20.7% in elderly women. Multivariate logistic regression analysis was used, after controlling the other covariates, to show that the significant related factors in hyperglycemia were obesity (odds ratio [OR] 2.1, 95% confidence interval [CI] 1.02-4.5), high systolic pressure (OR 2.1, 95% CI 1.1 4.0), and hypertriglyceridemia (OR 2.1, 95% CI 1.03-4.4). No significant association was found between hyperglycemia and gender, age, high diastolic pressure, abnormal glutamic pyruvic transaminase, hypercholesterolemia, hyperuricemia, renal function impairment, education level, retirement status, or marital status. CONCLUSIONS: The prevalence of hyperglycemia is high in elderly persons. Hyperglycemia is significantly associated with obesity, high systolic pressure, and hypertriglyceridemia in elderly persons. It is important to examine other cardiovascular risk factors if one cardiovascular risk factor is observed. PMID- 10819315 TI - Effects of chromium picolinate supplementation on insulin sensitivity, serum lipids, and body composition in healthy, nonobese, older men and women. AB - BACKGROUND: Chromium is an essential nutrient required for carbohydrate and lipid metabolism. Chromium supplementation in humans has been reported to improve glucose metabolism and improve serum lipid parameters and to reduce body fat; parameters that worsen with aging. As a result, chromium picolinate has been widely promoted as a health aid for the general population. The purpose of the study was to examine the effects of chromium supplementation on insulin sensitivity, serum lipids, and body composition in nonobese, healthy men and women of advanced age. METHODS: A randomized, double-blind, placebo-controlled study with 19 subjects (9 men and 10 women), aged 63-77, were given either chromium picolinate, 1,000 microg/d, or a placebo for 8 weeks. Serum lipids were measured at baseline and 8 weeks. Insulin sensitivity and body composition were measured with the minimal-model intravenous glucose tolerance test and dual energy x-ray absorptiometry scan, respectively, at baseline and after 8 weeks of chromium or placebo supplementation. RESULTS: No significant change in serum lipids, insulin sensitivity, or body composition was observed in the chromium group compared with the placebo group. CONCLUSIONS: Chromium picolinate supplementation alone does not appear to improve insulin sensitivity, serum lipids, or change body composition in nonobese, healthy men and women of advanced age. PMID- 10819316 TI - Association between complement regulatory protein factor H and AM34 antigen, detected in senile plaques. AB - BACKGROUND: We have previously shown that monoclonal antibody AM34, which is reactive with senile plaques, may recognize the C terminus of complement factor H. In this study, we investigated the expression of factor H in tissue from a human brain and the relation between AM34 antigen and factor H. METHOD: Total ribonucleic acid (RNA) was extracted from a normal human brain. A reverse transcriptase-polymerase chain reaction method was employed for detecting messenger RNAs coding for factor H and related proteins. Protein extracts from a normal human brain were also analyzed to detect factor H and related proteins by means of Western blotting. The cerebrospinal fluid from an Alzheimer's disease patient was immunoprecipitated with AM34 and anti-factor-H antibodies, and then it was subjected to gel electrophoresis followed by immunoblotting with AM34 and anti-factor-H antibodies. RESULTS: 26 clones of complementary DNA fragment were obtained by reverse transcriptase-polymerase chain reaction. Among them, seven clones were identical to factor H, and the others were related proteins and unreported sequences. A Western blot analysis of protein extracts from the normal brain tissue exhibited a 150-kd band, indicating the presence of factor H. AM34 was immunoreactive with the 150-kd molecule contained in the immunoprecipitates with anti-factor H antibodies, and vice versa. These results suggest that AM34 antigen could be identical to complement factor H. CONCLUSIONS: The results of our experiments indicate that factor H is possibly detected in the human brain, and that the AM34 antibody could recognize factor H. Because AM34 is capable of staining senile plaques positively, factor H is suggested to be associated with senile plaques in the human brain. PMID- 10819317 TI - Age-related differences in laterally directed compensatory stepping behavior. AB - BACKGROUND: Lateral falls are common in older adults and are associated with an elevated risk of hip fracture, compared with falls in other directions. Although rapid stepping movements can play an important functional role in maintaining balance, control of lateral stepping is a complex and demanding motor task. This study examined whether there are age-related differences in the stepping behavior used to recover from lateral loss of balance. METHODS: Rapid stepping reactions were evoked in healthy, active young (aged 20-30 years; N = 10) and older (aged 65-73 years; N = 10) volunteers by means of a sudden unpredictable motion of a platform on which the subject either stood quietly or walked in place. Subjects were instructed to respond naturally. Video analysis was performed to characterize the patterns of limb movement evoked by lateral platform motion. RESULTS: In responding to lateral perturbation of stance, the older adults were much more likely than the young adults to take multiple steps or use arm reactions to regain equilibrium, particularly when attempting crossover steps. During walk-in-place trials, both young and older subjects more frequently used a sequence of side steps rather than crossovers; however, older adults were still more likely to take extra steps or use arm reactions. Collisions between swing foot and stance limb occurred in 55% of walk-in-place trials in older adults versus only 8% in young adults. CONCLUSIONS: Control of lateral-stepping reactions appears to create difficulties for active and healthy older adults above and beyond previously reported problems in controlling forward and backward stepping. Impaired control of lateral-stepping reactions may be an early indicator of increased risk for lateral falls and hip fracture and should be an important consideration in the development of clinical approaches to predicting and preventing falls and related injuries. PMID- 10819318 TI - Factors associated with nursing-home entry for elders in Manitoba, Canada. AB - BACKGROUND: As the population ages, a greater demand for long-term care services and, in particular, nursing homes is expected. Policy analysts continue to search for alternative, less costly forms of care for the elderly and have attempted to develop programs to delay or prevent nursing-home entry. Health care administrators require information for planning the future demand for nursing home services. This study assesses the relative importance of predisposing, enabling, and need characteristics in predicting and understanding nursing-home entry. METHODS: Proportional hazard models, incorporating changes in needs over time, are used to estimate the hazard of nursing-home entry over a 5-year period, using health and sociodemographic characteristics of a representative sample of elderly residents from Manitoba, Canada. RESULTS: After age, need factors have the greatest impact on nursing-home entry. Specific medical conditions have at least as great a contribution as functional limitations. The presence of a spouse significantly reduces the hazard of entry for males only. CONCLUSIONS: The results suggest that the greatest gains in preventing or delaying nursing-home entry can be achieved through intervention programs targeted at specific medical conditions such as Alzheimer's disease, musculoskeletal disorders, and stroke. PMID- 10819319 TI - The prevalence of dizziness and its association with functional disability in a biracial community population. AB - BACKGROUND: Information regarding the prevalence of dizziness and its association with functional disability among African American and white residents from defined community populations is limited. METHODS: A total of 6,158 persons 65 years and older (78.8% of age-eligible persons) completed in-home interviews that included three common measures of self-reported disability: the Katz Activities of Daily Living (ADL) Scale, the Rosow-Breslau Functional Health Scale, and the Nagi Physical Disability Scale. A stratified random sample of 729 persons underwent a detailed evaluation that included questions on the frequency and severity of dizziness, which was defined as having an episode of dizziness or lightheadedness at least once a month. RESULTS: The overall prevalence of dizziness in this population was 9.6% (95% confidence interval [CI] 7.2-12.0). It increased with age, from 6.6% in those 65-74 years old, to 11.6% in those 75-84 years old, and to 18.4% in those persons > or =85 years old. It was more common in women (odds ratio [OR] 2.03, 95% CI 0.99-4.19) but was not associated with race. After adjusting for age, sex, and race, dizziness was associated with greater disability on the Rosow-Breslau (OR 2.29, 95% CI 1.18-4.46) and Nagi (OR 2.54, 95% CI 1.48-4.36) measures but not on the Katz ADL Scale (OR 1.18, 95% CI 0.64-2.20). CONCLUSIONS: Dizziness is common among older persons and is associated with functional disability. PMID- 10819320 TI - Age differences and changes in midline-crossing inhibition in the lower extremities. AB - BACKGROUND: The effects of crossing the body midline on the lower extremities of individuals 65 years of age and older was investigated. METHODS: The subjects were 10 individuals 65-79 years of age, 10 individuals 80 years and older, and 10 individuals 20-35 years. The total testing phase consisted of 2 sets of 30 trials per leg on 2 days. The subjects performed trials that involved movements ipsilaterally, contralaterally, and directly in front of the anterior superior iliac crest of the leg being tested. Reaction time and movement time scores were recorded. RESULTS: Individuals 65 years of age and older were found to exhibit slower reaction times to movements in a contralateral direction when compared with movements made in the ipsilateral direction whereas individuals 80 years of age and older were also found to exhibit slower reaction times to movements in a contralateral direction when compared with movements made in the midline and ipsilateral direction. CONCLUSIONS: These findings indicate that the effects of midline-crossing inhibition on the lower extremities reemerge in individuals 65 years and older, whereas in early development this effect disappears by 8 or 9 years of age. PMID- 10819321 TI - Prospective study of the impact of fear of falling on activities of daily living, SF-36 scores, and nursing home admission. AB - BACKGROUND: The aim of this study was to assess the impact of fear of falling on the health of older people. METHODS: A total of 528 subjects (mean age 77 years) were recruited from two hospitals in Sydney, Australia, and followed for approximately 12 months. Eighty-five subjects died during follow-up, and 31 were admitted to an aged care institution. Tinetti's Falls Efficacy Scale (FES) was successfully administered to 418 subjects as part of the baseline assessment. Among those with baseline FES scores, ability to perform 10 activities of daily living (ADLs) was assessed at baseline and follow-up in 307 subjects, and SF-36 scores were assessed at baseline and follow-up in 90 subjects recruited during the latter part of the study. Falls during follow-up were identified using a monthly falls calendar. RESULTS: Compared with those with a high fall-related self-efficacy (FES score = 100), those with a low fall-related self-efficacy (FES score < or = 75) had an increased risk of falling (adjusted relative risk 2.09, 95% confidence interval [CI] 1.31-3.33). Those with poorer fall-related self efficacy had greater declines in ability to perform ADLs (p < .001): the total ADL score decreased by 0.69 activities among persons with low FES scores (< or =75) but decreased by only 0.04 activities among persons with FES scores of 100. Decline in ADLs was not explained by the higher frequency of falls among persons with low FES scores. SF-36 scores (particularly scores on the Physical Function and Bodily Pain subscales) tended to decline more among persons with poor fall related self-efficacy. Nonfallers who said they were afraid of falling had an increased risk of admission to an aged care institution. CONCLUSIONS: Fear of falling has serious consequences for older people. Interventions that successfully reduce fear of falling and improve fall-related self-efficacy are likely to have major health benefits. PMID- 10819322 TI - Genomic, transcriptional and phenotypic analysis of ftsE and ftsX of Neisseria gonorrhoeae. AB - Although ftsE and ftsX are not universally present in bacteria, they are present in various Neisseria species as determined by Southern hybridization. The ftsE and ftsX genes of Neisseria gonorrhoeae (Ng) CH811 were cloned, sequenced and were shown to be co-transcribed from two promoters (P(E)1 and P(E)2) which were identified upstream of ftsE(Ng) by primer extension. Sequence analysis of FtsE(Ng) and alignment with other FtsE indicated that it contained the conserved motifs of ABC domains while sequence alignment of FtsX(Ng) with other published FtsX sequences predicted that they all contain four transmembrane segments and a conserved motif (Leu-hydrophobic aa-Gly-Ala/Gly) which may prove to be important for FtsX function. The viability of ftsE(Ng) and ftsX(Ng) mutants that were constructed by insertional inactivation indicated that these genes are not essential. The role of FtsE and FtsX is controversial. Analysis of ftsE(Ng) and ftsX(Ng) mutants by transmission electron microscopy showed that both exhibited morphological abnormalities indicative of defective division sites and in some cases aberrant condensation of DNA. PMID- 10819323 TI - In vivo conformation of mitochondrial DNA revealed by pulsed-field gel electrophoresis in the true slime mold, Physarum polycephalum. AB - Pulsed-field gel electrophoresis (PFGE) was used to examine the in vivo and in vitro conformations of Physarum polycephalum mitochondrial DNA (mtDNA). We used plugs containing isolated mitochondria, isolated mitochondrial nucleoids (mt nuclei), and isolated mtDNA, in addition to whole cells. The mtDNA contained in the myxamoebae, plasmodia, isolated mitochondria, and isolated mt-nuclei was circular, but most of the isolated mtDNA had been site-specifically fragmented and linearized during DNA preparation and storage under low ionic strength conditions. Restriction mapping of Physarum mtDNA by the direct digestion of the isolated mt-nuclei from two different strains, DP89 x AI16 and KM88 x AI16, resulted in the circular form. A linear mitochondrial plasmid, mF, is known to promote mitochondrial fusion and integration of itself into the mtDNA in Physarum. Linearization of mtDNA by the integration of the mF plasmid was demonstrated when we used PFGE to analyze isolated mitochondria from the plasmodial strain DP89 x NG7 carrying the mF plasmid (mF+). The PFGE system can be used not only to determine whether the form of mtDNA is linear or circular but also to analyze the dynamic conformational changes of mtDNA. PMID- 10819324 TI - Mapping of genes for cooking and eating qualities in Thai jasmine rice (KDML105). AB - Thai jasmine rice, KDML 105, is known as the best quality rice. It is known not only for its aroma but also for its good cooking and eating qualities. Amylose content (AC), gel consistency (GC) and gelatinization temperature (GT) are important traits determining rice quality. A population of recombinant inbred lines (RIL) derived from KDML105 x CT9993 cross was used to study the genetic control of AC, GC and GT traits. A total of 191 markers were used in the linkage map construction. The 1605.3 cM linkage map covering nearly the whole rice genome was used for QTL (define QTL) analysis. Four QTLs for AC were detected on chromosomes 3, 4, 6 and 7. These QTLs accounted for 80% of phenotypic variation explained (PVE) in AC. The presence of one major gene as well as several modifiers was responsible for the expression of the trait. Two QTLs on chromosome 6 and one on chromosome 7 were detected for GC, which accounts for 57% of PVE. A single gene of major effect along with modifier genes controls GC from this cross. The QTLs in the vicinity of waxy locus were major contributors in the expression of AC and GC. The finding that the position of QTLs for AC and GC were near each other may reflect tight linkage or pleiotropy. Three QTLs were detected, one on chromosome 2 and two on chromosome 6, which accounted for 67% of PVE in GT. Just like AC and GC, one major gene and modifier genes governed the variation in GT resulting from the KDML105 x CT9993 cross. Breeding for cooking and eating qualities will largely rely on the preferences of the end users. PMID- 10819325 TI - Characterization and identification of (CT)n microsatellites in soybean using sheared genomic libraries. AB - Three small insert (300 to approximately 600 bp) sheared genomic libraries were constructed by pipetting and DNase I treatment of soybean DNA. About 15,000 clones from each library were screened for CT- simple sequence repeats (CT-SSRs). The CT-SSRs were abundant in the soybean genome at an estimated frequency of approximately one SSR per 110 kb of genomic DNA. Following the sequencing of 129 positive clones, the repeat types and frequency of CT repeats among the positive clones were characterized. Forty-nine primer pairs were designed and preliminarily evaluated for their ability to amplify genomic DNA from a set of six varieties, including parents of a mapping family. Amplified products were analyzed by 10% PAGE. Eighty-eight percent of the designed primers were able to amplify all these genomic DNAs using a single PCR profile of 53 degrees C annealing temperature, of which 22 (45%) were polymorphic in the six varieties, and 14 of them were polymorphic in the parents of the mapping family. The polymorphic primer sets were further assessed for allelic information using DNA from 16 soybean cultivars. The average number of alleles was 4, ranging from 2 to 7 with the highest polymorphism information content value 0.84. Fourteen of these SSRs were mapped, using an existing soybean RFLP map. The findings presented here will advance our understanding of the soybean genome, and assist in the mapping genome and discrimination of closely related varieties of this species. PMID- 10819326 TI - Genomic characterization of the human heterotrimeric G protein alpha, beta, and gamma subunit genes. AB - Heterotrimeric guanine nucleotide binding proteins (G proteins) transduce extracellular signals received by transmembrane receptors to effector proteins. Each subunit of the G protein complex is encoded by a member of one of three corresponding gene families. Currently, 16 different members of the alpha subunit family, 5 different members of the beta subunit family, and 11 different members of the gamma subunit family have been described in mammals. Here we have identified and characterized Bacterial Artificial Chromosomes (BACs) containing the human homologs of each of the alpha, beta, and gamma subunit genes as well as a G alpha11 pseudogene and a previously undiscovered G gamma5-like gene. The gene structure and chromosome location of each gene was determined, as were the orientations of paired genes. These results provide greater insight into the evolution and functional diversity of the mammalian G protein subunit genes. PMID- 10819327 TI - C. elegans KLP-11/OSM-3/KAP-1: orthologs of the sea urchin kinesin-II, and mouse KIF3A/KIFB/KAP3 kinesin complexes. AB - Kinesins are intracellular multimeric transport motor proteins that move cellular cargo on microtubule tracks. It has been shown that the sea urchin KRP85/95 holoenzyme associates with a KAP115 non-motor protein, forming a heterotrimeric complex in vitro, called the Kinesin-II. Here we describe isolation of a cDNA clone corresponding to the klp-11 kinesin in C. elegans. Our sequence analysis of the encoded KLP-11 shows that it shares high homology with the OSM-3 kinesin. We also describe a nematode cDNA encoding KAP-1 that shares extensive homology with the sea urchin KAP115 kinesin associated protein. Sequence-based structural analysis of the OSM-3, KLP-11, and KAP-1, presented here suggests that these may form a heterotrimeric complex. We also describe the presence of a Drosophila armadillo consensus motif in CeKAP-1, first found in spKAP115, that suggests a possible role for the KAP-1 in signal transduction. PMID- 10819328 TI - Generation of 7137 non-redundant expressed sequence tags from a legume, Lotus japonicus. AB - For comprehensive analysis of genes expressed in a model legume, Lotus japonicus, a total of 22,983 5' end expressed sequence tags (ESTs) were accumulated from normalized and size-selected cDNA libraries constructed from young (2 weeks old) plants. The EST sequences were clustered into 7137 non-redundant groups. Similarity search against public non-redundant protein database indicated that 3302 groups showed similarity to genes of known function, 1143 groups to hypothetical genes, and 2692 were novel sequences. Homologues of 5 nodule specific genes which have been reported in other legume species were contained in the collected ESTs, suggesting that the EST source generated in this study will become a useful tool for identification of genes related to legume-specific biological processes. The sequence data of individual ESTs are available at the web site: http://www.kazusa.or.jp/en/plant/lotus/EST/. PMID- 10819329 TI - Structural analysis of Arabidopsis thaliana chromosome 3. I. Sequence features of the regions of 4,504,864 bp covered by sixty P1 and TAC clones. AB - Based on the physical map of Arabidopsis thaliana chromosome 3 previously constructed with CIC YAC, TAC, P1 and BAC clones (Sato, S. et al., DNA Res., 5, 163-168, 1998), a total of 60 P1 and TAC clones were sequenced, and the sequence features of the resulting 4,504,864 bp regions were analyzed by applying various computer programs for similarity search and gene modeling. As a result, a total of 1054 potential protein-coding genes were identified. The average density of the genes identified was 1 gene per 4066 bp. Introns were observed in 77% of the genes, and the average number per gene and the average length of the introns were 3.9 and 156 bp, respectively. These sequence features are essentially identical to those of chromosome 5 in our previous reports, but the gene density was slightly higher than that observed for chromosomes 2 and 4. The regions also contained 10 tRNA genes when searched by similarity to reported tRNA genes and the tRNA scan-SE program. The sequence data and information on the potential genes are available through the World Wide Web database KAOS (Kazusa Arabidopsis data Opening Site) at http://www.kazusa.or.jp/kaos/. PMID- 10819330 TI - Isolation and characterization of a novel zinc finger gene, ZNF219, and mapping to the human chromosome 14q11 region. AB - We have isolated and characterized a novel zinc finger gene by screening a human testis cDNA library. The isolated cDNA, termed ZNF219, contains an open reading frame of 2169 nucleotides encoding 723 amino acids. Sequence analysis revealed 9 sets of Kruppel-related zinc finger structures and proline-rich regions in several parts. ZNF219 exhibited ubiquitous expression in all fetal and adult tissues examined. The transcript size was 5.5 kb in adult tissues, while the main transcript size in the embryo stage was 3.5 kb. The transcript size is developmentally regulated. When the plasmid cloned with green fluorescent protein (GFP)-tagged ZNF219 was expressed in HeLa and COS7 cells, strong fluorescence intensities were observed only in the nucleus of both cells by fluorescence confocal microscopy. These data suggest that ZNF219 may be related to the regulation of transcription and developmental regulation. Genomic structure analysis mapped ZNF219 to chromosome 14q11 between markers D14S72 and D14S990, because a sequence tagged site mapped to the locus was found in the intron region of the ZNF219 gene. PMID- 10819331 TI - Prediction of the coding sequences of unidentified human genes. XVII. The complete sequences of 100 new cDNA clones from brain which code for large proteins in vitro. AB - To provide information regarding the coding sequences of unidentified human genes, we have conducted a sequencing project of human cDNAs which encode large proteins. We herein present the entire sequences of 100 cDNA clones of unknown human genes, named KIAA1444 to KIAA1543, from two sets of size-fractionated human adult and fetal brain cDNA libraries. The average sizes of the inserts and corresponding open reading frames of cDNA clones analyzed here were 4.4 kb and 2.6 kb (856 amino acid residues), respectively. Database searches of the predicted amino acid sequences classified 53 predicted gene products into the following five functional categories: cell signaling/communication, nucleic acid management, cell structure/motility, protein management and metabolism. It was also revealed that homologues for 32 KIAA gene products were detected in the databases, which were similar in sequence through almost their entire regions. Additionally, the chromosomal loci of the genes were determined by using human rodent hybrid panels unless their chromosomal loci were already assigned in the public databases. The expression levels of the genes were monitored in spinal cord, fetal brain and fetal liver, as well as in 10 human tissues and 8 brain regions, by reverse transcription-coupled polymerase chain reaction, products of which were quantified by enzyme-linked immunosorbent assay. PMID- 10819332 TI - A simple and efficient method to determine the terminal sequences of restriction fragments containing known sequences. AB - We present an improvement of the inverse PCR method for the determination of end sequences of restriction fragments containing unknown DNA sequences flanked by known segments. In this approach, a short "bridge" DNA is inserted during the self-ligation step of the inverse PCR technique. This bridge DNA acts as primer annealing sites for amplification and subsequent direct sequencing. Successive PCR amplifications enable selective amplification of the unknown sequences from a complex mixture. Unlike previously described methods, our method does not require special materials, such as synthetic adapters or biotinylated primers that must be prepared each time to adapt the target. Furthermore, no complex steps such as dephosphorylation or purification are needed. Our method can save time and reduce the cost of cloning unknown sequences; it is ideal for routine, rapid gene walking. We applied this method to a GC-rich bacterial genome and succeeded in determining the end sequences of a 4.5-kb fragment. PMID- 10819333 TI - Is the pneumococcus the one and only in acute otitis media? PMID- 10819334 TI - Critical review of evidence for short course therapy for tuberculous adenitis in children. PMID- 10819335 TI - Twice weekly vs. daily chemotherapy for childhood tuberculosis. AB - BACKGROUND: Treating childhood tuberculosis places a large burden on health services, and ways of lessening this were sought. METHODS: A randomized controlled trial was conducted to determine the effectiveness of fully intermittent twice weekly treatment for intrathoracic childhood tuberculosis and its effect on adherence to treatment, in comparison with daily (weekday) treatment. The setting was a district of Cape Town, South Africa, an area of high incident tuberculosis. We randomized 206 children with confirmed (4%), probable (94%) and suspected (2%) intrathoracic tuberculosis: 89 (median age, 25 months) received intermittent treatment; and 117 (median age, 28 months) received daily treatment. Intermittent treatment (twice weekly for 6 months) was isoniazid 15 mg/kg/dose, rifampin 15 mg/kg/dose and pyrazinamide 55 mg/kg/dose for 2 months, followed by isoniazid and rifampin only for 4 months. Daily treatment was isoniazid 10 mg/kg/day, rifampin 10 mg/kg/day and pyrazinamide 25 mg/kg/day on weekdays for 6 months. RESULTS: At 6 months 97% of subjects were discharged, with treatment outcomes in the two groups equivalent at that time (P = 0.90) and at the 18- to 30-month follow-up. One relapse occurred in the twice weekly group (P = 0.25). Adherence was equivalent; 70 children (79%) on intermittent and 90 (77%) on daily treatment took 75% or more of the prescribed doses (P = 0.90). Nonadherence over the full course of therapy was significantly associated with nonadherence during the first month of treatment (P = 0.0002) and household crowding (P = 0.002). CONCLUSIONS: Six month fully intermittent antituberculosis treatment is an effective and acceptable alternative to daily treatment. PMID- 10819336 TI - Zanamivir for treatment of symptomatic influenza A and B infection in children five to twelve years of age: a randomized controlled trial. AB - BACKGROUND: Influenza infection rates are higher in children than in other age groups. This study evaluated the efficacy, safety and tolerability of a 5-day course of twice daily inhaled zanamivir, 10 mg, compared with placebo in the treatment of symptomatic influenza A and B viral infections among children 5 to 12 years of age. METHODS: This double blind, randomized, placebo-controlled, parallel group, multicenter study conducted in the Northern Hemisphere during the 1998 and 1999 influenza season enrolled 471 patients with influenza-like symptoms for < or = 36 h. Patients were randomly assigned to zanamivir (n = 224) or placebo (n = 247). Symptoms were recorded on diary cards twice daily during treatment, for 9 days after treatment and for 14 additional days (if still reporting moderate/severe cough and/or taking relief medication). FINDINGS: A total of 346 (73%) patients were influenza-positive by culture, serology or polymerase chain reaction (65% influenza A, 35% influenza B). Zanamivir reduced the median time to symptom alleviation by 1.25 days compared with placebo among patients with confirmed influenza infection (P < 0.001). Zanamivir-treated patients returned to normal activities significantly faster and took significantly fewer relief medications than placebo-treated patients. Zanamivir was well-tolerated, demonstrating adverse event profiles similar to those of placebo and no clinically significant changes in laboratory findings. Viral susceptibility testing revealed no zanamivir-resistant strains of influenza A or B. CONCLUSIONS: Zanamivir was effective in shortening the duration and severity of influenza symptoms and was well-tolerated among children 5 to 12 years of age. PMID- 10819337 TI - Diminution of the anti-polyribosylribitol phosphate response to a combined diphtheria-tetanus-acellular pertussis/Haemophilus influenzae type b vaccine by concurrent inactivated poliovirus vaccination. AB - BACKGROUND: Prelicensure studies of Haemophilus influenzae type b vaccines (Hib) and diphtheria-tetanus-acellular pertussis vaccines (DTaP) were evaluated with concurrent oral poliovirus vaccine (OPV). However, inactivated poliovirus vaccine (IPV) is now recommended. A trial was conducted in which infants received a DTaP and Hib vaccine, separately (+) or combined (/), with either all OPV, all IPV or sequential IPV-OPV for the primary series of vaccinations. METHODS: In this protocol 567 infants were equally randomized to receive one of the following: Reference Arm A, DTaP + Hib + OPV; Treatment Arm B, DTaP/Hib + OPV; Treatment Arm C, DTaP/Hib + IPV at 2 and 4 months and OPV at 6 months; or Treatment Arm D, DTaP/Hib + IPV. antibodies against all administered antigens were measured at 7 months of age. Children with an antibody response to Hib (anti-polyribosylribitol phosphate (anti-PRP) <0.15 microg/ml had an antibody titer repeated after the toddler booster immunization. RESULTS: A significant diminution in the anti-PRP response was observed at 7 months of age in children given two or three doses of IPV concurrently with DTaP/Hib, compared with the groups given OPV. The geometric mean concentration of anti-PRP, percentage of children with > or = 0.15 microg/ml and percentage of children with > or = 1.0 microg/ ml, respectively, were: A, 4.4, 98%, 81%; B, 3.2, 94%, 78%; C, 1.3, 86%, 58% and D, 1.2, 84%, 53%. CONCLUSION: In this trial concurrent IPV appeared to interfere with the anti-PRP response to DTaP/Hib vaccine, suggesting that introduction of new vaccines may require evaluation of immune responses to all concurrently administered vaccines. PMID- 10819338 TI - Risk factors for mortality among children hospitalized because of acute respiratory infections in Bangui, Central African Republic. AB - BACKGROUND: Acute respiratory infections are the most common cause of death in children in developing countries. Little information is available on risk factors for mortality among African children presenting with symptoms compatible with acute respiratory infections. OBJECTIVE: To identify risk factors for death among children hospitalized for respiratory complaints who satisfy the WHO clinical definition for pneumonia or severe pneumonia. METHODS: Children <5 years of age who presented with cough and/or difficult breathing and were hospitalized in Bangui during a 1-year period were investigated for risk factors for mortality. The study population consisted of 395 children who satisfied the WHO clinical definition for pneumonia/severe pneumonia. The associations between death and demographic, nutritional, socioeconomic, laboratory and clinical variables were examined. RESULTS: Of the 49 (12.4%) children who died, all but one had had indrawing of the chest which, in univariate analysis, was the risk factor most strongly associated with death [odds ratio, 22.99; 95% confidence interval (CI), 3.81 to 935.2]. In a multivariate model the independent risk factors for death were indrawing of the chest [adjusted odds ratio (AOR) 8.35, CI 1.04 to 66.82], hepatomegaly (AOR 6.72, CI 2.35 to 19.21), age between 2 and 11 months (AOR 6.37, CI 2.18 to 18.59), grunting (AOR 4.53, CI 1.96 to 10.45), a moderate/severe alteration of general status (AOR 3.23, CI 1.17 to 8.94) and acute malnutrition (AOR 2.74, CI 0.96 to 7.78). CONCLUSIONS: These findings could be used in flow charts for the management of children with respiratory complaints to identify children at increased risk of death who need to receive aggressive therapy. PMID- 10819339 TI - Nonsusceptible Streptococcus pneumoniae in children with chronic otitis media with effusion and recurrent otitis media undergoing ventilating tube placement. AB - CONTEXT: Children with chronic otitis media are at risk for nonsusceptible Streptococcus pneumoniae (NSP) infection. If these children undergo ventilating tube placement, there is an opportunity to culture middle ear fluid and the nasopharynx to determine carriage of NSP. OBJECTIVE: To determine the incidence of NSP carriage, NSP antibiotic susceptibility and risk factors for NSP carriage in children with chronic otitis media undergoing tube placement. DESIGN AND SETTING: Prospective cohort study in an academic medical center with recruitment of patients from an otolaryngology private practice and clinic. PATIENTS: Children < 18 years of age undergoing tube placement for chronic otitis media. INTERVENTIONS: Myringotomy and tube placement, with culture of middle ear fluid and nasopharynx. MAIN OUTCOME MEASURES: The incidence of NSP cultured from the middle ears and nasopharynx of recruited subjects with the use of the minimum inhibitory concentration break points for penicillin susceptibility recommended by the National Committee for Clinical Laboratory Standards. RESULTS: S. pneumoniae was identified in at least 1 site from 23 of 300 study subjects (7.6%); of these 23, 12 case subjects (52.2%) harbored NSP. Of the risk factors assessed by preoperative questionnaire, only younger age was associated with NSP colonization (P < 0.0001). Of the six oral cephalosporins studied, cefpodoxime and cefuroxime showed good in vitro activity against S. pneumoniae isolates with intermediate penicillin resistance. CONCLUSIONS: Children with chronic otitis media undergoing tube placement may carry NSP and provide a means of monitoring the incidence of NSP and antibiotic susceptibilities for children with ear infections in their communities. Younger age is a risk factor for NSP carriage in this population. PMID- 10819340 TI - Antimicrobial resistance of nasopharyngeal isolates of Streptococcus pneumoniae and Haemophilus influenzae from children in the Central African Republic. AB - BACKGROUND: To assist the Central African Republic (CAR) develop national guidelines for treating children with pneumonia, a survey was conducted to determine antimicrobial resistance rates of nasopharyngeal isolates of Streptococcus pneumoniae (SP) and Haemophilus influenzae (HI). Secondary purposes of the survey were to identify risk factors associated with carriage of a resistant isolate and to compare the survey methods of including only children with pneumonia vs. including all ill children. METHODS: A cross-sectional survey of 371 ill children was conducted at 2 outpatient clinics in Bangui, CAR. RESULTS: In all 272 SP isolates and 73 HI isolates were cultured. SP resistance rates to penicillin, trimethoprim-sulfamethoxazole (TMP-SMX), tetracycline and chloramphenicol were 8.8, 6.3, 42.3 and 9.2%, respectively. All penicillin resistant SP isolates were intermediately resistant. HI resistance rates to ampicillin, TMP-SMX and chloramphenicol were 1.4, 12.3 and 0%, respectively. The most common SP serotypes/groups were 19, 14, 6 and 1; 49% of HI isolates were type b. History of antimicrobial use in the previous 7 days was the only factor associated with carriage of a resistant isolate. Resistance rates were similar among ill children regardless of whether they had pneumonia. CONCLUSIONS: Resistance rates were low for antimicrobials recommended by the World Health Organization for children with pneumonia. We recommended TMP-SMX as the first line treatment for pneumonia in CAR because of its low cost, ease of dosing and activity against malaria. PMID- 10819341 TI - Effect of fever on the serum antibody response of Gambian children to Haemophilus influenzae type b conjugate vaccine. AB - BACKGROUND: Acute malaria is a major pediatric problem in developing countries and it is known to be immunosuppressive. METHODS: The serum antibody response to Haemophilus influenzae type b (Hib) conjugate vaccine was investigated in children ages 12 to 30 months with fever associated with malaria, fever associated with other causes or no fever. Groups of 57 children with malaria, 57 children with fever without malaria and 60 healthy children were bled and vaccinated with a single dose of H. influenzae type b capsular polysaccharide tetanus protein conjugate vaccine. Of these 137 were bled again 1 to 2 months after vaccination. RESULTS: The median antibody titers at baseline were low and similar in the three groups; 77, 65 and 57% of children in the malaria, febrile and healthy groups, respectively, had prevaccination titers of anti polyribosylribitol phosphate antibodies below 0.15 microg/ml. The median antibody titers after vaccination were 6.3, 7.5 and 23 microg/ml in the malaria, febrile and healthy groups, respectively (P < 0.001, healthy group vs. the two febrile groups). All the healthy children had protective titers (>0.15 microg/ml) after vaccination, but 11% of the children with malaria and 4% of the other febrile children did not have protective titers. CONCLUSIONS: Anti-polyribosylribitol phosphate titers after Hib vaccination were lower in children with malaria or other febrile illnesses at the time of vaccination than in controls. Fever associated with malaria or other acute illnesses is associated with a diminished response to Hib conjugate vaccine. These findings raise questions about the vaccination of febrile children and indicate the need for further studies in this area. PMID- 10819342 TI - Ciprofloxacin for treatment of tularemia in children. AB - BACKGROUND: Children with tularemia are, irrespective of severity of disease, usually subjected to parenteral treatment with aminoglycosides. Based on available susceptibility testing, quinolones might be effective oral alternatives of parenteral therapy. These drugs cause arthropathy in immature animals, but this risk is currently regarded to be low in humans. PATIENTS AND METHODS: In 12 patients (median age, 4 years; range, 1 to 10) with ulceroglandular tularemia, a 10- to 14-day course of oral ciprofloxacin, 15 to 20 mg/kg daily in 2 divided doses, was prescribed. Microbiologic investigations included identification of the infectious agent by PCR and culture of wound specimens, as well as determination of antibiotic susceptibility of isolates of Francisella tularensis. RESULTS: Defervescence occurred within 4 days of institution of oral ciprofloxacin in all patients. After a median period of 4.5 days (range, 2 to 24), the patients were capable of outdoor activities. In 2 cases, treatment was withdrawn after 3 and 7 days because of rash. In both cases a second episode of fever occurred. All children recovered without complications. In 7 cases F. tularensis was successfully cultured from ulcer specimens and tested for susceptibility to ciprofloxacin. MIC values for all isolates were 0.03 mg/l. CONCLUSION: In our sample of 12 patients ciprofloxacin was satisfactory for outpatient treatment of tularemia in children. PMID- 10819343 TI - Pneumococcal bacteremia during a decade in children in Soweto, South Africa. AB - OBJECTIVES: To monitor for a decade the incidence and the clinical and microbiologic characteristics of pneumococcal bacteremia in children in Soweto and to assess the influence of HIV infection on any changes. METHODS: Case records of children with pneumococcal bacteremia at Chris Hani Baragwanath Hospital from July, 1986, to June, 1987 (1986/ 1987), and from July, 1996, to June, 1997 (1996/ 1997), were retrospectively reviewed. RESULTS: There were 194 episodes, 62 in 19861 1987 and 132 in 1996/1997. The minimum annual incidence for children younger than 5 years of age increased from 61 per 100000 (179 per 100000 for those <12 months old) in 1986/1987 to 130 per 100000 (349 per 100000 for those <12 months old) in 1996/1997. Sixty-seven (60%) of 111 patients tested in 1996/1997 were HIV-seropositive; none were tested in 1986/1987. The HIV-infected compared with HIV-noninfected were more likely to be malnourished (61% vs. 36%, P = 0.02), less likely to have other underlying disease (12% vs. 50%, P = 0.00001) and more frequently used antibiotics recently (69% vs. 43%, P = 0.008). Penicillin-nonsusceptible isolates were found in 22 (35%) patients in 1986/1987 and 52 (39%) in 1996/1997. There was no significant change in antimicrobial susceptibility during the decade or by HIV serostatus. CONCLUSIONS: Children in Soweto had a high incidence of pneumococcal bacteremia which doubled during the decade mainly as a result of the impact of the HIV epidemic. There has been no significant change in antimicrobial susceptibility for the decade. PMID- 10819344 TI - Five vs. ten days of antibiotic therapy for acute otitis media in young children. AB - BACKGROUND: Many publications in recent years have argued in favor of shortened therapy for acute otitis media. However, doubt persists regarding children younger than 2 years, and some authors therefore restrict short course therapy to children older than 2 years. METHODS: In a prospective, comparative, double blind, randomized, multicenter trial we compared cefpodoxime-proxetil, 8 mg/kg/day in two divided doses for 10 days, with an identical 5-day regimen followed by a 5-day placebo period. RESULTS: Between October, 1996, and April, 1997, 450 children (mean age, 14.3 months) were enrolled, 227 in the 5-day group and 223 in the 10-day group. In the per protocol analysis clinical success was obtained on Days 12 to 14 after the beginning of treatment (main analysis) in 175 (84.1%) of the 208 children receiving the 5-day regimen and 194 (92.4%) of the 210 children receiving the 10-day regimen (P = 0.009). The superiority of the standard regimen was more marked among children cared for outside their homes (92.5% vs. 81.5%). Clinical success persisted on Days 28 to 42 among 134 (85.4%) of the 157 assessable patients in the 5-day group and 144 (83.7%) of the 172 assessable patients in the 10-day group (P = 0.68). CONCLUSIONS: The 10-day regimen resulted in a higher success rate at the conclusion of therapy, but there were no differences between the two study groups 4 to 6 weeks after enrollment in the study protocol. PMID- 10819345 TI - Safety and immunogenicity of a nonavalent pneumococcal vaccine conjugated to CRM197 administered simultaneously but in a separate syringe with diphtheria, tetanus and pertussis vaccines in Gambian infants. AB - BACKGROUND: Unrelenting high morbidity and mortality have mandated that immunogenic vaccines be used to combat pneumococcal disease in infants. OBJECTIVES: To evaluate the safety and immunogenicity of a nonavalent pneumococcal conjugate vaccine and the antigenic interaction when administered simultaneously with diphtheria, tetanus and pertussis vaccines. METHODS: Two hundred seven infants were randomized to receive three doses of either nonavalent protein conjugate pneumococcal vaccine (PnCV) or inactivated polio vaccine (IPV) at 2, 3 and 4 months of age with routine Expanded Program of Immunization vaccines as scheduled. Vaccinees were visited on Days 1, 2 and 7 to observe local and systemic adverse reactions. Blood was drawn before the first dose and 1 month after the third dose. Antibody concentrations in sera were measured by standardized enzyme-linked immunosorbent assay. Nasopharyngeal carriage of pneumococci was tested at 5 and 9 months of age. RESULTS: No serious reactions were observed. Local induration and tenderness were observed more commonly at the site of administration of diphtheria, tetanus and pertussis vaccines than at the site of administration of IPV or PnCV. Between 79 and 91% achieved >1 microg/ml antibody against specific pneumococcal serotypes. Antibody responses to diphtheria and pertussis antigens were similar in both groups; however, antibody response to tetanus toxoid was significantly lower in infants who received PnCV (geometric mean concentration, 11.1 vs. 17.4; P < 0.001). Nasopharyngeal carriage in PnCV-vaccinated children was reduced but not significantly different from those vaccinated with IPV. CONCLUSION: Simultaneous administration of PnCV with Expanded Program of Immunization vaccines is safe and immunogenic. immune response to the composite antigens is likely to confer protection. PMID- 10819346 TI - Duration of therapy for acute otitis media. PMID- 10819347 TI - Expanding microbiology of pulmonary infection in cystic fibrosis. PMID- 10819348 TI - Progressive respiratory distress in an infant treated for presumed pertussis. PMID- 10819349 TI - Tuberculosis-associated hemophagocytic syndrome in an infant. PMID- 10819350 TI - Lack of association between Kawasaki syndrome and infection with parvovirus B19, human herpesvirus 8, TT virus, GB virus C/hepatitis G virus or Chlamydia pneumoniae. PMID- 10819351 TI - Acute disseminated histoplasmosis with multifocal choroiditis in a child. PMID- 10819352 TI - Intrauterine staphylococcal scalded skin syndrome: report of a case. PMID- 10819353 TI - Primary cutaneous aspergillosis in ventilated neonates. PMID- 10819354 TI - Mycobacterium fortuitum osteomyelitis of the cuboid after nail puncture wound. PMID- 10819355 TI - Tungiasis in a young child adopted from South America. PMID- 10819356 TI - Bacillus anthracis sepsis in a newborn. PMID- 10819357 TI - Monitoring of aminoglycoside serum concentrations. PMID- 10819358 TI - Echovirus 30 outbreak of aseptic meningitis in Turkey. PMID- 10819359 TI - Utility of peripheral venous blood cultures in patients with central venous catheters. PMID- 10819360 TI - High altitude as an explanation for bronchiolitis-associated hospitalizations. PMID- 10819361 TI - Multiple nodal basin drainage in truncal melanomas. PMID- 10819362 TI - Sentinel node metastasis from thin melanomas with vertical growth phase. PMID- 10819363 TI - Extended (D2) lymph node dissection for gastric cancer: do patients benefit? PMID- 10819364 TI - Significance of multiple nodal basin drainage in truncal melanoma patients undergoing sentinel lymph node biopsy. AB - BACKGROUND: Although previous studies have demonstrated that truncal site is associated with an adverse prognosis, explanations for such risk are lacking. In addition, the number of nodal basins as well as the number of lymph nodes containing regional metastases are important prognostic factors in these patients. Because the lymphatic drainage pattern of truncal melanoma often includes more than one basin, we designed a study to evaluate (1) whether patients with multiple nodal basin drainage (MNBD) were at an increased risk of lymph node metastases identified by sentinel lymph node (SLN) biopsy, and (2) whether the histological status of an individual basin reliably predicted the status of the other draining basins in patients with MNBD. METHODS: The records of 295 consecutive truncal melanoma patients who were managed primarily with intraoperative lymphatic mapping and SLN biopsy, between 1991 and 1997, were reviewed. All patients underwent preoperative lymphoscintigraphy, which established the number and location of draining nodal basins. Univariate and multivariate analyses of relevant clinicopathological factors were performed to assess which factors may predict the presence of a pathologically positive SLN. RESULTS: At least one SLN was identified in 281 patients. MNBD was present in 86 (31%) patients, and a pathologically positive SLN was found in 56 (20%) patients. By multivariate analysis, the presence of MNBD (relative risk = 1.9; P = .03), tumor thickness (P = .007), and tumor ulceration (relative risk = 2.4; P = .01) were significant independent risk factors for the presence of at least one pathologically positive SLN. SLN pathology in one basin did not predict the histology of other basins in 19 (22%) of 86 patients with MNBD. CONCLUSIONS: MNBD is independently associated with an increased risk of nodal metastases in truncal melanoma patients. Because the histological status of an individual basin did not reliably predict the status of the other draining basins in patients with MNBD, it is important to adequately identify and completely assess all nodal basins at risk, as defined by lymphoscintigraphy, in truncal melanoma patients. PMID- 10819365 TI - Incidence of sentinel node metastasis in patients with thin primary melanoma (< or = 1 mm) with vertical growth phase. AB - BACKGROUND: Patients with thin primary melanomas (< or = 1 mm) generally have an excellent prognosis. However, the presence of a vertical growth phase (VGP) adversely impacts the survival rate. We report on the rate of occurrence of nodal metastasis in patients with thin primary melanomas with a VGP who are offered sentinel lymph node (SLN) biopsy. METHODS: Among 235 patients with clinically localized cutaneous melanomas who underwent successful SLN biopsy, 71 had lesions 1 mm or smaller with a VGP. The SLN was localized by using blue dye and a radiotracer. If negative for tumor by using hematoxylin and eosin staining, the SLN was further examined by immunohistochemistry. RESULTS: The rate of occurrence of SLN metastasis was 15.2% in patients with melanomas deeper than 1 mm and 5.6% in patients with thin melanomas. Three patients with thin melanomas and a positive SLN had low-risk lesions, based on a highly accurate six-variable multivariate logistic regression model for predicting 8-year survival in stage I/II melanomas. The fourth patient had a low- to intermediate-risk lesion based on this model. At the time of the lymphadenectomy, one patient had two additional nodes with metastasis. CONCLUSIONS: VGP in a melanoma 1 mm or smaller seems to be a risk factor for nodal metastasis. The risk of nodal disease may not be accurately predicted by the use of a multivariate logistic regression model that incorporates thickness, mitotic rate, regression, tumor-infiltrating lymphocytes, sex, and anatomical site. Patients with thin lesions having VGP should be evaluated for SLN biopsy and trials of adjuvant therapy when stage III disease is found. PMID- 10819366 TI - Systemic toxicity and cytokine/acute phase protein levels in patients after isolated limb perfusion with tumor necrosis factor-alpha complicated by high leakage. AB - BACKGROUND: Since the introduction of high-dose tumor necrosis factor-alpha (TNFalpha) in the setting of isolated limb perfusion (ILP) in the clinic, prevention of leakage to the body of the patient is monitored with great precision for fear of TNF-mediated toxicity. That we observed remarkably little toxicity in patients with and without leakage prompted us to determine patterns of cytokines and acute phase proteins in patients with high leakage and in patients without any leakage. METHODS: TNFalpha, interleukin (IL)-6, IL-8, C reactive protein, and secretory (s)-phospholipase A2 were measured at several time points during and after (until 7 days) ILP in 10 patients with a leakage to the systemic circulation varying in percentage from 12% to 65%. As a control, the same measurements, both in peripheral blood and in perfusate, were performed in nine patients without systemic leakage. RESULTS: In patients with systemic leakage, levels of TNFalpha increased during ILP, reaching values to 277 ng/ml. IL-6 and IL-8 peaked 3 hours after ILP with values significantly higher compared with patients without systemic leakage. C-reactive protein and s-phospholipase A2 peaked at day 1 in both patient groups, s-phospholipase A2 with significant higher levels and C-reactive protein, in contrast, with lower levels in the leakage patients. CONCLUSIONS: High leakage of TNFalpha to the systemic circulation, caused by a complicated ILP, led to 10-fold to more than 100-fold increased levels of TNFalpha, IL-6, and IL-8 in comparison with patients without leakage. The increase of the acute phase proteins was limited. Even when high leakage occurs, this procedure should not lead to fatal complications. The most prominent clinical toxicity was hypotension (grade III in four patients), which was easily corrected. No pulmonary or renal toxicity was observed in any patient. It is our experience that, even in the rare event of significant leakage during a TNFa-based ILP, postoperative toxicity is usually mild and can be easily managed by the use of fluid and, in some cases, vasopressors. PMID- 10819367 TI - Diagnostic primer sets for microsatellite instability optimized for a minimal amount of damaged DNA from colorectal tissue samples. AB - BACKGROUND: The diagnosis of microsatellite instability from a minimal amount of highly damaged DNA, extracted from formalin-fixed, paraffin-embedded tissue by the microdissection method, is difficult. Therefore, optimized primer sets were newly designed for substitution of documented ones. METHODS: DNA was extracted from 15 archival colorectal carcinomas and used as templates for polymerase chain reaction. Nine standard microsatellite markers (BAT-25, BAT-26, BAT-40, D18S69, D2S123, D5S346, D10S197, D17S250, and D18S58) were selected for diagnosis of microsatellite instability in colorectal carcinomas. All polymerase chain reaction conditions for primer sets were unified to save experimental time. RESULTS: The primer sets for the latter five markers documented in the literature were redesigned because of poor efficiency for damaged DNA. As a result, the number of DNA samples, sufficiently amplified at all markers, improved from 0% to 93%. CONCLUSIONS: Diagnostic primer sets for microsatellite instability, optimized for a minimal amount of damaged DNA from colorectal tissue samples, were established. PMID- 10819368 TI - Preoperative multivariate prediction of morbidity after gastrectomy for adenocarcinoma. AB - BACKGROUND: Gastrectomy remains the only curative treatment for gastric cancer. However, surgical morbidity and mortality remains high. Our aim was to identify the risk factors that determine operative morbidity and mortality and to describe a simple method for preoperative stratification of morbidity outcome. METHODS: Retrospective review of patients who underwent gastrectomy for gastric cancer. Multivariate analysis was used to define risk factors for surgical morbidity and mortality. RESULTS: A total of 208 cases were included. Fifty-one episodes of operative morbidity and 19 surgery-related deaths were found. Operative blood loss (risk ratio [RR], 1.0012), serum albumin (RR, 0.42), extent of gastrectomy (RR, 2.8), lymphocyte count (RR, 0.999), and splenectomy (RR, 1.51) were the most important risk factors for morbidity. However, location of the tumor, serum albumin level, and lymphocyte count were the most important preoperative risk factors that determine the appearance of surgical complications. Receiver operating characteristic analysis of this model allowed definition of three risk groups in terms of surgical morbidity (11.8%, 28.5%, and 52.4%, respectively). CONCLUSIONS: A new method for preoperative calculation of the probability of surgical complications was developed. It must be validated prospectively and in different settings to be used in preoperative interventions designed to reduce that risk. PMID- 10819369 TI - Differences in release mechanisms and distributions for sialyl Le(a) and sialyl Le(x) antigens in colorectal cancer. AB - BACKGROUND: To investigate colorectal cancer-related carbohydrate antigen release and distribution, we evaluated serum levels of sialyl Le(a) (CA19-9) and sialyl Le(x) antigen (SLX) in blood samples obtained from both a peripheral vein and a tumor's draining vein. METHODS: Blood samples were obtained during surgery from 126 patients. Based on these samples, patients were placed into a high-antigen group, with a concentration above a selected cutoff value, or into a low-antigen group, with a tumor marker concentration below that same value. The blood samples obtained from peripheral veins were designated by the "p" prefix, and samples from drainage veins were designated by the "d." RESULTS: Serum d-SLX levels were significantly higher than p-SLX levels (P < .0001), although there was no difference between those of d-CA19-9 and p-CA19-9. Only 1 (3.6%) of 28 patients in the high d-CA19-9 group had a low p-CA19-9. In contrast, 6 (33.3%) of 18 patients in the high d-SLX group had low p-SLX levels (P = .0103). Correlations between pathological variables and either p-CA19-9 levels or d-CA19-9 levels were similar. However, both distant metastasis and venous invasion did prove to be independent variables related to d-SLX levels, as shown by logistic regression analysis. CONCLUSIONS: SLX may drain predominantly via the draining veins of colorectal tumors into portal circulation, whereas CA19-9 may drain via another route. PMID- 10819370 TI - Hemodynamic and cardiac function parameters during heated intraoperative intraperitoneal chemotherapy using the open "coliseum technique". AB - BACKGROUND: Heated intraoperative intraperitoneal chemotherapy achieves high peritoneal concentrations with limited systemic absorption and has become an important tool in the management of patients with peritoneal carcinomatosis from low-grade malignancies such as pseudomyxoma peritonei and in selected cases of high-grade tumors such as colon adenocarcinoma. When the closed abdomen technique is used, its perioperative toxicity seems to be related to the hemodynamic and cardiac function changes associated with increased body temperature and increased intra-abdominal pressure. METHODS: Hemodynamic and cardiac function variables during heated intraoperative intraperitoneal chemotherapy, using an open abdomen "coliseum technique," were measured in 15 patients with the use of a noninvasive esophageal Doppler monitor. RESULTS: The hemodynamic and cardiac function changes were characterized by an increased heart rate, increased cardiac output and decreased systemic vascular resistance associated with an increased body temperature, and decreased effective circulating volume with the urinary output tending to decrease as the therapy progressed. CONCLUSION: Heated intraoperative intraperitoneal chemotherapy with the open abdomen coliseum technique induces a hyperdynamic circulatory state with an increased intravenous fluid requirement and avoids changes because of increased intra-abdominal pressure. Hemodynamic and cardiac stability, as documented by normal blood pressure and adequate urinary output, can be achieved by liberal intravenous fluids, titrated to frequent urinary output determination. PMID- 10819371 TI - PCNA and Ki-67 as prognostic markers in human parathyroid carcinomas. AB - BACKGROUND: It is widely accepted that histological diagnosis of parathyroid tumors is established with great difficulty. Carcinomas cannot be reliably separated from adenomas by histology alone. In this study, immunohistochemical staining for proliferating cell nuclear antigen (PCNA) and Ki-67 was determined in 10 cases of parathyroid carcinomas, labeling indices (LIs) were calculated, and the results were correlated with the clinical outcomes. METHODS: Ten cases of formalin-fixed, paraffin-embedded tissue with surgically resected parathyroid carcinoma were used. Immunohistochemical staining for PCNA and Ki-67 was performed and the LIs were calculated. We also examined whether LI could become a useful marker for parathyroid carcinomas. RESULTS: Although nine patients with minimally invasive growth without recurrence of the tumor showed a low LI for both markers, one patient with a widely invasive neoplasm, and who died, had a high LI. CONCLUSIONS: These results suggested that the LI of PCNA and Ki-67, in addition to the histological appearance, may be markers of the biological behavior of parathyroid carcinomas. However, this study was on a small scale, so it may be valuable to repeat these studies in a larger group of patients with better defined histological criteria. PMID- 10819372 TI - Prognostic significance of immunohistochemically localized biomarkers in stage II and stage III breast cancer: a multivariate analysis. AB - BACKGROUND: The aim was to investigate the expression of a panel of biomarkers such as prolactin (PRL), p53, Bcl-2, c-erb B2, Ki-67, CD44, and factor VIII related antigen (FVIII-RA) in primary tumors of stage II and stage III breast cancer and its correlation with disease prognostication. METHODS: The streptavidin-biotin peroxidase complex technique was used for the detection of these antigens. Cytoplasmic staining pattern was observed for PRL, Bcl-2, and Ki 67. Staining pattern for p53 was nuclear. Membranous and/or cytoplasmic staining was noted for c-erb B2 and CD44. Microvessel staining was noted for FVIII-RA. RESULTS: Of the 93 primary breast tumors analyzed, positivity for PRL was noted in 82%, for p53 in 56%, for Bcl-2 in 73%, for c-erb B2 in 68%, and for Ki-67 and CD44 in 78% each. The microvessel count (MVC) for FVIII-RA ranged from 0.0 to 29.0, with a median of 6.0, which was used as a cutoff. MVC > or = 6.0 was noted in 51% of breast tumors. With increasing tumor size, the higher frequency of positivity of MVC > or = 6.0 (P = .0001), CD44 (P = .001), PRL (P = .002), and c erb B2 (P = .008), and higher frequency of Bcl-2 negativity (P = .033), was noted. In stage III patients, a higher positivity of the following biomarkers was noted, compared with stage II patients: MVC > or = 6.0 (P = .0004), PRL (P = .0002), c-erb B2 (P = .001), and CD44 (P = .005). Further, Bcl-2 positivity was significantly lower in patients with stage III disease compared with those with stage II disease (P = .024). In patients with nodal involvement, the frequency of c-erb B2 (P = .006), MVC > or = 6.0 (P = .011), and PRL (P = .032) was higher than in those without nodal involvement. Moreover, in these patients, with the increase in the number of involved lymph nodes, there was a significant increase in frequency of CD44+ (P = .0004) and PRL+ (P = .013) tumors. Abnormal expression of one biomarker was seen in 7% of tumors, of two biomarkers in 4%, of three in 15%, of four in 19%, of five in 28%, of six in 20%, and of all seven biomarkers in 7% of tumors. The frequency of an increasing number of biomarkers coexpressed was higher in stage III patients compared with stage II patients (P = .00003). In the total number of patients (n = 93), tumors with Bcl-2 negativity (P = .00001), MVC > or = 6.0 (P = .001), PRL positivity (P = .02), and CD44 positivity (P = .034) had a significantly poorer overall survival (OS) compared with their respective counterparts. In stage II patients (n = 40), only p53 expression was significantly associated with reduced relapse-free survival (P = .009) and OS (P = .040). In multivariate analysis, p53 expression was an independent prognostic factor that influenced relapse-free survival (P = .034) of stage II breast cancer patients. However, it failed to attain statistical significance for OS. In stage III patients (n = 53), tumors with Bcl-2 negativity (P = .0005) and MVC > or = 6.0 (P = .039) had a significantly poorer OS compared with their respective counterparts. In multivariate analysis of stage III patients, Bcl-2 was the only independent prognostic factor (P = .001) for predicting OS. There was a significant association between coexpression of the biomarkers and OS (P = .001). The OS rates decreased with the increase in number of abnormally expressed biomarkers. CONCLUSIONS: p53 expression in primary tumors was an independent prognostic factor that influenced relapse-free survival in patients with stage II disease. In stage III patients, lack of Bcl-2 expression was independently associated with a poor prognosis and, thus, may be an indicator of aggressive phenotype. PMID- 10819373 TI - Resection of invasive pulmonary aspergillosis in immunocompromised patients. AB - BACKGROUND: Immunocompromised patients are prone to develop invasive pulmonary aspergillosis (IPA). Relapse and high mortality rates are seen in those patients who receive subsequent immunotoxic therapy. Standard antifungal regimens often fail to completely eradicate IPA, which then warrants an aggressive surgical approach. METHODS: We performed a retrospective chart review of 13 immunocompromised patients who were considered to have IPA and who underwent surgery between 1988 and 1998. RESULTS: Twelve patients had a hematological malignancy and one patient had breast cancer. The diagnosis of IPA was based on a chest computed tomographic scan in all patients. A preoperative diagnosis of aspergillosis was made in three patients, and mucormycosis in one patient, by bronchoalveolar lavage. Before surgery, seven patients received chemotherapy, one patient underwent bone marrow transplantation, and five patients received a combination of chemotherapy and bone marrow transplantation. Symptoms included cough (54%), fever (54%), hemoptysis (30%), and shortness of breath (8%). Three patients (23%) were asymptomatic. The mean preoperative absolute neutrophil count was 4881 cells/microl. Seventeen thoracic operations were performed, i.e., 12 wedge resections, 4 lobectomies, and 1 pneumonectomy. One patient also underwent nephrectomy for invasive aspergillosis and one patient underwent craniotomy to resect an aspergillus brain mass. Surgical pathology revealed IPA in 13 (76%), invasive mucormycosis in 2 (15%), aspergilloma in 1, and diffuse alveolar hemorrhage in 1. Postoperative complications included the following: operative bleeding requiring transfusion, three patients; prolonged air leak, two patients; death because of hepatic/renal failure, one patient; and death because of overwhelming multisystem aspergillosis, one patient. Seven (54%) patients underwent further immunotoxic treatment with no aspergillosis recurrence. After a mean follow-up of 12 months, five (38%) patients are alive and seven (54%) have died without evidence of aspergillosis and/or mucormycosis. CONCLUSIONS: Surgical resection, in combination with antifungal agents, is a safe and effective form of therapy for invasive mycoses. It prevents recurrence and allows for subsequent cytotoxic therapies. PMID- 10819374 TI - Is intensive follow-up really able to improve prognosis of patients with local recurrence after curative surgery for rectal cancer? PMID- 10819375 TI - A review of telemedicine in accident and emergency: the story so far. AB - Recent developments in information and communications technology have the potential to revolutionise health care. This has been recognised at government level, and plays a significant part in the new information strategy for the NHS "Information For Health". Telemedicine (literally, medicine at a distance) is one of the most successful techniques in this rapidly expanding field, and in preliminary studies has proved to be both successful and popular with patients and health care professionals. In the UK telemedicine has been mainly applied to two major areas of accident and emergency (A&E) practice. These are the transmission of computed tomography scans for urgent neurosurgical opinion and the ongoing support of minor injuries units. The latter also involves transmission and interpretation of radiographs, usually peripheral limb films. Telemedicine is not a medical subspecialty in itself, but a facilitator of all medical and surgical specialties. While recent modernisation initiatives have permitted A&E departments to purchase a range of telemedical equipment, overall progress is hampered by a lack of large or scientifically rigorous studies, and a complete absence of data on the economic implications of this new technique. This review introduces A&E telemedicine in terms that avoid jargon and complex technical details. After a brief consideration of the origins of the subject, attention is given to recent publications relating to minor injuries support and A&E teleradiology. The technical and clinical feasibility of A&E telemedicine are demonstrated, and a case is made for the transmission and interpretation of minor injuries radiographs using a relatively simple and inexpensive system, supported by timely radiological reporting. After a brief study of various legal and ethical issues, the likely developments of the future are discussed. PMID- 10819376 TI - Current opinions and practices in the treatment of spontaneous pneumothorax. AB - The approach to the initial management of spontaneous pneumothorax differs markedly from centre to centre, and it is difficult in practice to establish a standard protocol. This article reviews the concepts behind the British Thoracic Society guidelines, and reports the varying opinions and alternative practices existing currently. There is a need for more evidence-based studies to identify what is the best approach. Based on a review of relevant recent reports, the author attempts to work out an unbiased practical approach that can be used safely and that can possibly give the best overall cost effective results. PMID- 10819377 TI - Echoes of things to come. Ultrasound in UK emergency medicine practice. AB - Ultrasound is widely used in the US and continental Europe in the immediate assessment of patients after blunt abdominal trauma. There are also now other recognised "primary" indications for ultrasound in emergency medicine. In this paper current evidence supporting the implementation and use of emergency ultrasound in these primary conditions and possible other indications are assessed. The issues surrounding introduction of the technology into the practice of emergency medicine in this country are considered. It is accepted that further debate is necessary but the establishment of a robust evidence base in the UK will help to clarify the place of ultrasound. PMID- 10819378 TI - Brain attack!--How good is the early management of subarachnoid haemorrhage in accident and emergency departments? South Thames A&E Specialty Sub Committee Audit Group. AB - OBJECTIVES: The aim of this study was to assess the characteristics and audit the management of patients for whom a diagnosis of subarachnoid haemorrhage was made and who were admitted to a neurosurgical centre from an accident and emergency (A&E) department. The objective was to use the results to make recommendations for improving care in this group of patients. METHODS: Four neurosurgical centres in the South Thames Region provided lists of patients admitted with a diagnosis of subarachnoid haemorrhage during 1997. The medical records and computed tomography of 162 patient episodes from 20 A&E departments were reviewed. The speed of presentation, the provisional diagnosis and the referral patterns were examined. Patient records were audited to investigate what proportion were treated according to established guidelines while in the A&E departments. RESULTS: 63.7% of patients presented to A&E within six hours of ictus and 81.1% within three days. Some 49.2% of patients were seen by a doctor within 30 minutes and 74.7% within one hour. Computed tomography was performed on 18.8% within two hours and only 66.3% within six hours of arrival at the A&E department. Eighteen per cent were treated with nimodipine, 26% with intravenous fluids and 32% were given analgesia before leaving the department. Delays occurred in obtaining computed tomography. CONCLUSIONS: The management of patients who present to A&E departments with subarachnoid haemorrhage is suboptimal. Improved awareness of the disorder, management by senior A&E staff and clear care pathways could help with management. PMID- 10819379 TI - The effect of a picture archiving and communications system (PACS) on diagnostic performance in the accident and emergency department. AB - OBJECTIVE: A study has been conducted to identify the benefits to the accident and emergency (A&E) department of a hospital wide Picture Archiving and Communications System (PACS). METHODS: The study was conducted in two parts: firstly while the hospital was using conventional radiographic films, and secondly when the PACS was in operation. For each part of the study, the diagnoses of radiographic images made by A&E clinicians were compared with those made by radiologists. This resulted in the estimation of the incidence of false negative findings by the A&E staff. The management of patients with such findings was studied to identify those for whom a change of treatment was required. Such data for the two periods, when film and when PACS was used, were compared. RESULTS: It was found that the overall rate of misdiagnoses across all A&E patients who had radiography was low in both periods and there was a significant reduction when PACS was used (1.5% for film and 0.7% for PACS, 95% CI for difference between proportions: -0.014 to -0.0034), but the rate of serious misdiagnoses involving patient recall did not change significantly (95% CI for difference between proportions: -0.0059 to +0.0001). CONCLUSIONS: When PACS was used the diagnostic performance by A&E staff improved by reducing false negative interpretations but the rate of serious misdiagnosis did not change. PMID- 10819380 TI - A process approach to improving pain management in the emergency department: development and evaluation. AB - AIM: (1) To describe a process approach to the improvement of pain management in emergency departments. (2) To compare analgesia ordering and administration practices for patients with acute fractures before and after implementation of a nurse managed, titrated intravenous narcotic policy. METHOD: Retrospective chart review of patients with long bone fractures for the years 1993 and 1997. RESULTS: There was a dramatic change in analgesia administration practices away from the intramuscular route in favour of the intravenous route (p<0.001). For long bone fractures, in 1993, 53% of patients received intramuscular narcotic analgesia compared with 5% in 1997. In contrast, in 1993, 6% of the patients received intravenous narcotic analgesia compared with 54% in 1997. CONCLUSION: This study demonstrates that a process approach to improving pain management that resulted in both changes in drug administration and pain assessment and management processes made a significant and sustained change to analgesia ordering and administration practices for patients with long bone fractures in an emergency department. PMID- 10819382 TI - Emergency department discharge instructions: a wide variation in practice across Australasia. AB - OBJECTIVES: All patients discharged from the emergency department (ED) should be given discharge instructions (DIs). This study aimed to describe the DI practices of EDs in Australasia and to examine the differences between public and private EDs. METHODS: A voluntary, questionnaire-based survey of public and private EDs throughout Australia and New Zealand. RESULTS: 58 of 74 (78.4%) EDs provided instruction notes upon discharge although 51 (68.9%) gave them to only "some" patients. There were no significant differences between the public and private EDs (p>0.05). Thirty seven (50%) EDs provided pre-formatted instruction sheets although 22 (29.7%) EDs gave them to only "some" patients. There were no significant differences between the public and private EDs (p>0.05). Only five (6.8%) EDs retained a copy of the instruction sheets and no (O%) ED provided sheets that included all recommended features. Sixty six (89.2%) EDs provided information sheets although 44 (59.5%) EDs gave them to only "some" patients. Private EDs gave information sheets to significantly more patients than public EDs (p=0.04). CONCLUSIONS: The DI practices of individual EDs varied and the rates of DI provision were low. EDs should improve their DI practices. Pre formatted instruction sheets, containing all recommended features, should be provided to all patients with a copy kept in the medical record. Consideration should be given to the use of DI practices as an ED performance indicator. PMID- 10819381 TI - The management of hyperkalaemia in the emergency department. AB - Life threatening hyperkalaemia (> 7.0 mmol/l) is commonly associated with acute renal failure. Moderate hyperkalaemia (6.1-6.9 mmol/l) is also common and well tolerated in patients with chronic renal failure. Renal failure is the most common cause of hyperkalaemia although other causes to consider include drugs (potassium sparing diuretics, angiotensin converting enzyme inhibitors), hyperglycaemia, rhabdomyolysis and adrenal insufficiency. Hyperkalaemia affects the cardiac conducting tissue and can cause serious arrhythmias including ventricular fibrillation and asystolic arrest. Therefore it is important to treat hyperkalaemia promptly in the emergency department. This paper evaluates the therapeutic options available for treatment of hyperkalaemia. PMID- 10819383 TI - The development of an assault patient questionnaire to allow accident and emergency departments to contribute to Crime and Disorder Act local crime audits. AB - OBJECTIVE: To evaluate and refine an assault patient questionnaire to facilitate the contribution of accident and emergency (A&E) departments to Crime and Disorder Act local crime audits. METHOD: A brief nine item questionnaire was devised in collaboration with the authors of the Home Office British Crime Survey. A prospective sample of 46 consecutive assault patients who attended Cardiff Royal Infirmary A&E department were interviewed by either reception staff or triage nurses. The questionnaire was revised appropriately. RESULTS: The collection of information in A&E departments about the circumstances of violence was straightforward. Questions about motive for violence and about relationships between the injured and their assailants were problematic. CONCLUSION: The collection of information relevant to Crime and Disorder Act crime audits was possible without extra resource. Receptionists were found to be the most appropriate staff to record information. PMID- 10819384 TI - The domestic iron. A danger to young children. AB - OBJECTIVES: To study the epidemiology of thermal injury caused by the domestic iron in children 5 years old or less. METHODS: Retrospective review of case notes held in the accident and emergency (A&E) department of a large teaching hospital over a 36 month period. Data regarding demographics, site and extent of injury, mechanism of injury and outcome were retrieved. RESULTS: 62 thermal injuries were identified in 59 patients. Of these, 60 were contact burns and two were scalds. The male to female ratio was 2:1. The mean age was 24 months. Fifty five per cent were aged between 1 and 2 years old. The hand was the commonest site of injury (63%) and, of these, two thirds were on the palm. Interestingly 10% occurred on the face. Iron contact burns accounted for 23.5% of all contact burns in this age group over this period. The majority of contact burns were partial thickness and most were less than 1% body surface area. Inadequate supervision is a recurring theme in many of these cases. A suspicion of non-accidental injury was raised in 10 cases and confirmed in nine of these. CONCLUSIONS: Iron burns are common in young children, particularly boys aged between 1 and 2 years old. Most can be treated in the A&E clinic. The potential for serious injury does exist. Non accidental injury always needs to be considered. Efforts at prevention and increasing public awareness are needed. PMID- 10819385 TI - Physiological variables during open chest cardiopulmonary resuscitation: results from a small series. AB - OBJECTIVES: To evaluate the efficacy of open chest cardiac compressions for the resuscitation of pre-hospital cardiac arrest patients presenting with non shockable rhythms on arrival at the emergency department. DESIGN: Prospective observational study of outcomes and physiological parameters during open chest cardiac compressions for non-traumatic pre-hospital cardiac arrest. SETTING: Large accident and emergency department receiving pre-hospital cardiac arrest patients with pre-hospital advanced life support provided by ambulance service paramedics. SUBJECTS: All patients in whom open chest cardiac compressions were performed by the author (JCC) during the period from January to May 1998 (seven patients). INTERVENTIONS: Assessment of artificial pulse and blood pressure generated by open chest cardiac compressions, measurement of arterial blood gases, recovery of spontaneous cardiac output, recovery of spontaneous ventilatory efforts and survival to admission and to discharge. RESULTS: Artificial pulse and recordable non-invasive blood pressures were generated in all patients; PO2 was physiological or supra-physiological in all patients; PCO2 was physiological or subphysiological in all patients; pH and base deficit were not corrected in the five patients with repeated samples (including two receiving 50 mEq sodium bicarbonate); three patients recovered spontaneous cardiac output (ROSC); two patients recovered spontaneous respiratory efforts (unrelated to ROSC); no patients survived to admission. CONCLUSIONS: Open chest cardiac compressions provide effective perfusion, enabling correction of ventilation parameters and showing clinical signs of adequate perfusion. However, acidosis was not corrected and the use of 50 mEq sodium bicarbonate was ineffective. PMID- 10819386 TI - An introduction to everyday statistics--1. AB - Descriptive statistics are used to summarise numerical information so that it is in a more manageable form. There are a variety of ways of carrying this out depending upon what type of data we are dealing with. There is also a choice when presenting data. Graphical and tabular formats are possible but each have strengths and weaknesses. Selection therefore needs to take these into account along with the format of the presentation and the type of data. PMID- 10819387 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Antibiotics and compound finger fracture. PMID- 10819388 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Troponin T to rule out myocardial damage in chest pain. PMID- 10819389 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Flumazenil and suspected benzodiazepine overdose. PMID- 10819390 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Corticosteroids in acute spinal cord injury. PMID- 10819391 TI - Fracture of the occipital condyle: the forgotten part of the neck. AB - A case of occipital condylar fracture in a multiply injured and unconscious motorcyclist is reported. This injury was clinically unsuspected but found on the lowest cuts of head computed tomography. It is shown that this site is often inadequately imaged when scanning the head and neck in victims of trauma. The Anderson and Montesano classification of occipital condylar fracture is described. It is noted that types 1 and 2 are stable injuries but type 3 is potentially unstable. A retrospective analysis of 30 head computed tomography scans in trauma cases revealed that in only 16 were the occipital condyles adequately imaged. It is emphasised that vigilance is required to detect fractures of the occipital condyle and that it should be standard practice to include this area when performing computed tomography of the head in trauma victims. PMID- 10819392 TI - Spontaneous pneumothorax: is it under tension? AB - A diagnosis of tension pneumothorax is usually only considered within the context of trauma, incorrect chest drain insertion or positive pressure ventilation. Four patients are presented who developed spontaneous tension pneumothorax with no precipitating factors. In three of these instances, the diagnosis was only made radiologically and in every case the treating physician was unaware that a spontaneous tension pneumothorax could occur. Previously, emphasis has been placed on tracheal deviation in a tension pneumothorax. However, this is an inconsistent finding as one of the cases highlights. Patients may appear surprisingly clinically well until they decompensate. These cases are highlighted to raise awareness of this potentially life threatening condition. PMID- 10819393 TI - Aorto caval fistula--the "bursting heart syndrome". AB - Aorto caval fistula is one of the less well recognised complications of abdominal aortic aneurysm seen in accident and emergency departments. It presents in a number of different ways the commonest of which is high output congestive cardiac failure with warm peripheries. Initial diagnosis is based on the index of suspicion of the clinician. However, early diagnosis by the emergency physician and early surgery can markedly improve the patients prognosis. PMID- 10819394 TI - Screwdriver assaults and intracranial injuries. AB - Four patients with intracranial penetrating injuries from screwdrivers are presented. Two cases were fatal; the others were left with functional deficits. In two of the patients a penetrating injury was not suspected initially because the history was limited and the significance of the small entry wounds were not appreciated. Unless these wounds are carefully examined a penetrating injury is easily overlooked. PMID- 10819395 TI - Volar metacarpophalangeal joint dislocation. AB - Volar dislocation of the metacarpophalangeal joint is a very rare clinical finding. A volar dislocation of the metacarpophalangeal joint of the left index finger in a 44 year old man is reported. Closed reduction proved unsuccessful requiring subsequent open reduction and internal fixation via a combined dorsal and volar approach. The presentation, mechanism of injury and treatment of this case and other previously reported cases are discussed. PMID- 10819396 TI - Spontaneous spinal epidural haematoma: an unusual cause of neck pain. PMID- 10819397 TI - Late presentation of diaphragmatic hernia and gastric volvulus. PMID- 10819398 TI - Self insertion of urethral foreign bodies. PMID- 10819399 TI - An unusual cause of pilonidal sinus. PMID- 10819400 TI - Myocardial infarction and left bundle branch block. PMID- 10819401 TI - Pedestrian injuries sustained in negotiating traffic calming measures. PMID- 10819402 TI - Shock and ipsilateral pulmonary oedema. PMID- 10819403 TI - Reproducibility of time-domain and three different frequency-domain techniques for the analysis of the signal-averaged electrocardiogram. AB - Because time-domain (TD) analysis of the signal-averaged ECG (SAECG) has some limitations that limit its use, several frequency-domain analysis techniques were developed in an attempt to improve the diagnostic ability of the SAECG. However, it is not known how reliable these techniques are at detecting late potentials. This prospective study compares the short-term reproducibility of 4 analysis techniques: TD analysis, spectral temporal mapping (STM), spectral turbulence analysis (STA), and acceleration spectrum analysis (ASA), in a large series of normal patients and post-myocardial infarction (MI) patients. Two consecutive SAECGs were recorded in 634 patients that were divided into 3 groups: 117 remote MI patients undergoing programmed electrical stimulation for the inducibility of ventricular tachycardia (Group 1), 407 consecutive acute MI survivors (Group 2), and in 110 healthy volunteers (Group 3). The diagnostic reproducibility of the 4 techniques was evaluated by comparing rates of inconsistent results (1 normal and the other abnormal). The numeric reproducibility for each technique was assessed by comparing the normalized differences of each single SAECG parameter between the 2 recordings. Inconsistent results of diagnostic reproducibility were observed in 4.1%, 6.9%, 9.8%, and 18.0%, with TD, STA, ASA, and STM, respectively. Comparisons of these rates were significantly different (P < .05) except between STA and ASA (P = .07). The numeric reproducibility was highest for TD parameters, lowest for STM factors of normality, and intermediate for STA and ASA indices. TD analysis remains the most reproducible SAECG analysis technique, whereas STM showed the worst reproducibility, which limits its clinical applicability. STA and ASA provide an acceptable intermediate reproducibility, the former being slightly, although not significantly, more reproducible than the latter. PMID- 10819404 TI - The short-term and long-term reproducibility of spectral turbulence and late potential variables of the signal-averaged ECG in a population sample of healthy subjects and the impact of gender, age, and noise. AB - Previous methods for frequency domain analysis of the signal-averaged ECG (SAECG) have had low reproducibility. The reproducibility of time domain late potential analysis and spectral turbulence analysis was evaluated with 2 immediately consecutive SAECG recordings in 121 randomly selected subjects without heart disease (short-term) and also in 47 subjects after 1 month (long-term). A test was late potential positive if 2 or more of 3 variables were outside the reference limits and spectral turbulence positive if the score was 3 or 4. The short-term reproducibility was high for the filtered QRS duration (FQRSD), root mean square amplitude of the last 40 ms (RMS40) and high frequency low amplitude signals less than 40 microV (HFLAS40) of the time domain and total QRS duration (TQRSD), power spectral density of the last 40 ms and the late potential duration of time domain analogous analyses. The Spearman rank order correlation coefficients were 0.89, 0.88, and 0.84 and 0.97, 0.91 and 0.97, respectively. The reproducibility of the spectral score variables varied, and the correlation for the low slice correlation ratio was 0.71, spectral entropy 0.61, interslice correlation mean 0.58, and interslice correlation SD 0.28. A diagnostic inconsistency between 2 tests occurred in 0 (0%) subjects in late potential analysis if FQRSD was required for positivity, and in 7 (6%) otherwise, and in 9 (7%) of spectral turbulence analysis. If the spectral variable mean peaks per slice, with a correlation of 0.89, replaced interslice correlation SD in the spectral score, diagnostic inconsistency occurred in 0 (0%) subjects. The reproducibility seemed higher in women and in younger people but significantly only for interslice correlation subjects mean and HFLAS40. The long-term reproducibility did not differ significantly from short-term for any variable. In conclusion, the reproducibility was high in all time domain and time domain analogous variables. It varied among the spectral turbulence score variables and was very low for interslice correlation SD. The reproducibility of the spectral score improved substantially if this variable was replaced by mean peaks per slice. PMID- 10819405 TI - Beat-to-beat QT interval variability in coronary patients. AB - Changes in ventricular repolarization have been described in patients after myocardial infarction, whereas data for coronary patients without prior myocardial infarction are lacking. This study was designed to evaluate ventricular repolarization in coronary patients with effort angina pectoris. Beat to-beat QT interval variability (QTV) using 5-minute resting high-resolution ECG recordings was measured in 26 men (mean age 62.1 years) with effort angina pectoris and without prior myocardial infarction, and in 30 age-matched men without clinically evident coronary heart disease (controls). To evaluate the degree of coronary artery disease in coronary patients, coronary angiography was performed. Coronary patients displayed significantly higher values of QTV compared with the control patients (P < .001). Rate adaptation of QT interval correlated significantly with the degree of coronary artery disease in the study group patients (P < .05). The significant association between QTV and coronary heart disease suggests altered ventricular repolarization in coronary patients without prior myocardial infarction. PMID- 10819406 TI - Recurrent ischemia during continuous 12-lead ECG-ischemia monitoring in patients with acute coronary syndromes treated with eptifibatide: relation with death and myocardial infarction. PURSUIT ECG-Ischemia Monitoring Substudy Investigators. Platelet glycoprotein IIb/IIIa in Unstable angina: Receptor Suppression Using Integrilin Therapy. AB - Computer-assisted continuous monitoring of the ST-segment allows detection and quantification of recurrent ischemia in patients with acute coronary syndromes. In a substudy of the PURSUIT (Platelet glycoprotein IIb/IIIa in Unstable angina: Receptor Suppression Using Integrilin Therapy) trial, this technique was used to evaluate the effects of the glycoprotein IIb/IIIa inhibitor eptifibatide on the incidence and severity of recurrent ischemia, and to investigate the relationship between recurrent ischemia and the occurrence of subsequent death or myocardial (re)infarction. A total of 258 patients with unstable angina or evolving myocardial infarction without ST elevation were monitored for 24 hours during infusion with either eptifibatide or placebo with a computer-assisted 12-lead ECG ischemia monitoring device. Recurrent ischemic episodes were identified by an automated computer algorithm. Two hundred and sixteen patients (84%) had ECG recordings suitable for analysis. Ischemic episodes were detected in 35 (33%) of the 105 eptifibatide patients and in 32 (29%) of the 111 placebo patients (not significant). No difference in ischemic burden was apparent between both treatment groups. Patients who exhibited 2 or more episodes of recurrent ischemia more frequently died or suffered a myocardial infarction, both at 7 and 30 days, as well as through the 6-month follow-up. A greater ischemic burden was significantly related to adverse outcome during the 6-month follow-up period. Real-time computer-assisted continuous multilead ECG-ischemia monitoring may help to identify patients with unstable coronary syndromes at increased risk of adverse outcome and, thus, allow for better prognostic triage and more appropriate selection of therapeutic strategies. Integration of these systems in coronary care units and emergency wards should, therefore, be recommended. PMID- 10819407 TI - Enhancing long-term ECG monitoring with graphic analysis of coupling intervals. AB - By graphically identifying patterns of ventricular ectopic beat (VEB) interval characteristics, we sought to enhance arrhythmia analysis, especially in long term ECG monitoring. Coupling intervals as a function of preceding sinus RR intervals (CI/RR diagrams) with the aid of coupling interval and interectopic interval histograms were analyzed in 172 patients with frequent VEBs. Four distinct types of CI/RR diagrams were observed: horizontal-elongated (25 patients), linear (4 patients), triangular (37 patients), and clusters separated by dot-sparse areas (17 patients). In 89 patients, no definite pattern was discerned. The patients with elongated diagrams were significantly younger, had fixed coupling, uniform QRS complexes, and lacked periodicities in their interectopic interval histograms. The linear pattern was detected in a small group with rate-related VEBs. In the group with a triangular pattern, 30% of the patients exhibited interectopic periodicities suggesting parasystole. The patients with dot-sparse areas in the CI/RR diagrams had more variable coupling and predominantly more multiform QRS complexes. We conclude that CI/RR diagrams in conjunction with coupling interval and interectopic interval histograms enhance arrhythmia analysis by identifying patterns, such as those consistent with either fixed coupling, rate dependence, parasystole, or multiform VEBs. PMID- 10819408 TI - Frequency-domain heart rate variability in 24-hour Holter recordings: role of spectral method to assess circadian patterns and pharmacological autonomic modulation. AB - Different spectral methodologies for heart rate variability were recently shown to provide the same qualitative results in the context of passive tilt test. However, the impact of the method and the use of normalized power units in long term ECG monitoring is still debated. Autoregressive and Fast Fourier transform (FFT) spectral approaches were applied to assess circadian modulation and the effect of beta-blocker administration in mild hypertensive patients who underwent continuous ambulatory ECG recording (n = 44, 51 +/- 12 years, 30 men). Spectral analysis was applied to 5-minute sequences and spectral parameters representative of each circadian period (24 hour, day, night) were calculated. In baseline recordings, FFT spectral method provided a smaller estimate of total and very low frequency powers. On the contrary, low- and high-frequency components were systematically larger with FFT. Circadian variations were in favor of an increased overall nocturnal variability but of a reduced low frequency normalized power with both spectral methods. Chronic oral administration of beta-blocker induced an increase of all spectral components except for an unchanged low frequency normalized power, independently from the spectral approach. In spite of quantitative differences, the qualitative assessment of circadian patterns and beta-blockade effect by autoregressive- and FFT-based spectral analyses is equivalent. The low-frequency component of heart rate variability cannot be considered a reliable direct marker of sympathetic activity in long-term ambulatory ECG recording. PMID- 10819409 TI - Rapid ventricular repolarization in rodents: electrocardiographic manifestations, molecular mechanisms, and clinical insights. AB - This article examines specific electrocardiographic (ECG) and electrophysiological features of ventricular repolarization in rats and mice, and the role of depolarization-activated potassium currents in mediating the unique features of ECG recordings in these rodents. This article describes the currents that underlie ventricular repolarization in these rodents, identifies terminology that appropriately describes the unique features of murine ECG recordings, and correlates these unique findings with selected human ECG ventricular repolarization abnormalities. The absence of a distinct isoelectric interval between the QRS complex and the T wave, accompanied by a relatively short QT interval, are common features of ECG recordings in mice and rats, but not in ECGs in guinea pigs. The murine ECG morphology is apparently attributable to the presence of large outward K+ currents that dominate the early phase of ventricular repolarization. In rats and mice, the predominant current underlying the early phase of repolarization appears to be the rapidly activating and inactivating 4-aminopyridine-sensitive transient outward current (ie, I(to)). Importantly, the density of I(to) in rats and mice is high, whereas this current is not evident in the ventricular myocytes of guinea pigs. The high density of I(to) appears to underlie the prominent J wave or downsloping ST-segment elevation seen in rats and mice, whereas the ST-segment is isoelectric in guinea pigs. The unusual J wave and ST-segment pattern in murine ECGs, however, does bear some resemblance to ECG features observed in humans with Brugada syndrome, and with hypothermia and ischemia. These patterns in rats and mice might, therefore, serve as an experimental model for the idiopathic J wave. PMID- 10819410 TI - Estimates of repolarization and its dispersion from electrocardiographic measurements: direct epicardial assessment in the canine heart. AB - This study investigates a technique to estimate dispersion based on the root mean square (RMS) signal of multiple electrocardiographic leads. Activation and recovery times were measured from 64 sites on the epicardium of canine hearts using acute in situ or Langendorff perfused isolated heart preparations. Repolarization and its dispersion were altered by varying cycle length, myocardial temperature, or ventricular pacing site. Mean and dispersion of activation and recovery times, and activation-recovery interval (ARI) were calculated for each beat. The waveform was then calculated from all leads. Estimates of mean and dispersion of activation and recovery times and mean ARI were derived using only inflection points from the RMS waveform. QT intervals were also measured and QT dispersion was determined. Estimates determined from the RMS waveform provided accurate estimates of repolarization and were, in particular, a better measure of repolarization dispersion than QT dispersion. PMID- 10819411 TI - Influences of catecholamines on the sudden death induced in dogs by an antifungal agent, D0870. AB - We previously reported the occurrence of QT prolongation and sudden death owing to torsades de pointes (TdP) in dogs treated with D0870, an antifungal agent. In the present study, we evaluated the influences of epinephrine and isoproterenol on the onset of TdP each time D0870 was given to 6 anesthetized open-chest dogs at a dosage of 20 mg/kg, 5 times every 40 minutes, by the simultaneous measurements of surface electrocardiogram and epicardial monophasic action potential (MAP). D0870 alone induced noticeable prolongation of the QT interval and action potential duration (APD), but neither ventricular premature contraction (VPC) nor sudden death. In contrast, the additional administration of the catecholamines induced a greater shortening of APD during the later phase of repolarization than during its earlier one and VPCs in all dogs tested, and sudden deaths owing to TdPs in 4 of the 6 dogs treated with D0870. These results suggest that D0870 alone does not induce TdP but that catecholamines play an important part in the development of sudden death induced by D0870 in dogs. PMID- 10819412 TI - Incomplete ECG expression of acute true posterior myocardial infarction, owing to an antecedent anterior infarction. AB - We present a 75-year-old man with an inferior and true posterior (TP) myocardial infarction (MI), who showed disproportionately deeper ST-segment depression (decreased ST) in the precordial electrocardiogram (ECG) leads than the ST segment elevation (increased ST) noted in leads 2, 3, and aVF. This suggested that the precordial decreased ST was not reciprocal to the inferior increased ST, but it was indicative of TP ischemic injury. However, the precordial decreased ST, in subsequent serial ECGs, was not followed up by R waves of increasing amplitude, or tall T waves in the V1 and V2 leads, as would be expected had this been a case of evolving TP MI. Nevertheless, TP MI was confirmed by echocardiography and dipyridamole/thallium SPECT myocardial perfusion scintigraphy. These last 2 modalities also revealed evidence of an anterior (A) MI, which had indeed occurred 11 years previously, and was confirmed by a history of hospitalization, serial ECGs, and enzymatic evidence of necrosis. This incomplete ECG expression of TP MI in our patient was felt to be owing to the previous large A MI, which had long ago deprived the heart from requisite healthy myocardium opposite the recent TP MI, for generation of tall R waves or R/S ratio greater than 1 in the V1 and V2 leads. This case is presented as an example of electrical cancellation affecting the QRS complexes. PMID- 10819413 TI - Structure of the reentrant circuit of idiopathic left ventricular tachycardia: new insights into the role of the Purkinje network. AB - In idiopathic left ventricular tachycardia (ILVT), the reentrant circuit is considered to involve the Purkinje system, and the Purkinje potential (P potential) appears to be a marker for successful ablation. However, the characteristics of the reentrant circuit in ILVT have not yet been defined. In 2 cases of ILVT, we performed detailed mapping along the left ventricular septum during VT and sinus rhythm. ILVTs were successfully ablated at the posteroapical area of the left ventricular septum where the high frequency P-potential was recorded and this portion was considered to be the exit site of the reentrant circuit. A small P-potential was also recorded at the portion proximal to the exit site, and it preceded the P-potential at the exit site. However, the local ventricular electrogram at the exit site preceded that at the proximal site during VT. Moreover, the small P-potential was orthodromically entrained by ventricular pacing from the proximal site. These findings suggest that the reentry circuit of ILVT appeared to have considerable size. PMID- 10819414 TI - New onset left bundle branch block with right axis deviation in a patient with Wegener's granulomatosis. AB - Left bundle branch block (LBBB) is usually associated with a normal axis or left axis deviation (LAD). When it is seen in association with right axis deviation (RAD) it is felt to be a marker of diffuse advanced myocardial disease. We report a case of new onset LBBB with RAD in a patient with Wegener's granulomatosis who had an otherwise functionally and structurally normal heart. To our knowledge, this is the first case report of LBBB with RAD without severe cardiomyopathy, as well as the first case report of new onset LBBB as a result of Wegener's granulomatosis. PMID- 10819415 TI - Differentiation of atrial flutter from atrial fibrillation. PMID- 10819416 TI - An enhanced process for encapsulating aspirin in ethyl cellulose microcapsules by solvent evaporation in an O/W emulsion. AB - An enhanced process for microencapsulating aspirin in ethylcellulose was demonstrated using an oil-in-water emulsification/solvent evaporation technique. Methylene chloride (CH2Cl2) was used as the dispersed medium and water as the dispersing medium. The recovered weight, particle size distribution, aspirin loading efficiency, and the aspirin release rate of microcapsules were analysed. The addition of appropriate amounts of non-solvent (n-heptane) prior to the emulsification increases the recovered weight, but decreases the size of the formed microcapsules. The addition of non-solvent also changes the microcapsule characteristics, resulting in a coarser surface and an increased release rate. Increasing the polymer (ethylcellulose) concentration in the dispersed phase increases the size of the microcapsules, the recovered weight, and loading efficiency, but decreases the release rate. The release rate follows first-order kinetics during the first 12 h, suggesting a monolithic system with aspirin uniformly distributed in the microcapsule. PMID- 10819417 TI - Calcium pectate gel beads for cell entrapment. 6. Morphology of stabilized and hardened calcium pectate gel beads with cells for immobilized biotechnology. AB - The structure of standard and stabilized calcium pectate gel (CPG) beads has been examined by scanning (SEM) and transmission (TEM) electron microscopy. A two stage crosslinking procedure with polyethyleneimine (PEI) and glutaraldehyde (GA) led to the formation of a more compact layer on the bead surface. On the other hand, the stabilization procedure did not significantly change either gel bead interior or morphologic properties, vitality and biotransformation activity of immobilized bacterial cells (Nocardia tartaricans) against cis-epoxysuccinate as well as yeast cells (Trigonopsis variabilis) against cephalosporin C. The structure of these cells within the calcium pectate matrix remained unchanged. Moreover, the two-step chemical stabilization of CPG containing T. variabilis or N. tartaricans had a favourable effect on storage and operational stability at semi-continuous and continuous processing in stirred batch and packed-bed reactors. The most valuable effect of stabilization was the fact that the hardened CPG comprising the cells N. tartaricans resisted, for a long time (360 days and more), the destructive effects of the product (such strong sequestering reagent as L-(+)-tartaric acid) at high concentrations (up to 1 M). Non-hardened CPG was destroyed after 21 h. The reference materials, hardened and non-hardened calcium alginate gels (CAG), were destroyed over 3 h or 30 min, respectively. PMID- 10819418 TI - Comparison of in vivo fate and immunogenicity of hepatitis B surface antigen incorporated in cationic and neutral liposomes. AB - To compare cationic liposomes (CatL) and neutral liposomes (NeuL), as a vaccine carrier, the in vivo fate and immunogenicity of hepatitis B surface antigen (HBsAg), incorporated in CatL and NeuL, were investigated. CatL, composed of phosphatidyl choline (PC) and stearyl amine (SA) with a molar ratio of 9:1, showed a 2.5-fold higher incorporating efficiency of HBsAg than NeuL composed of PC alone. Most of HBsAg incorporated in both liposomes existed in an antibody available form on the outer surface of liposomes. After intramuscular injection to rats, HBsAg in CatL resided at the injection site for a longer period than that in NeuL with terminal half lives of 52.5 and 42.9 h, respectively. However, HBsAg in NeuL was more efficiently taken up by the lymphatic organs and spleen than that in CatL. Furthermore, the group treated with HBsAg in NeuL showed earlier sero-conversion with higher anti-HBsAg titre than the group treated with HBsAg in CatL. Sero-conversion rates (SCRs) in both CatL- and NeuL-treated animals were 100% after every injection carried out, except the primary injection of CatL. These results demonstrate that CatL can enhance the retention of incorporated antigen at the injection site, compared with NeuL. However, the production of antibody by HBsAg in NeuL is more effective than that by HBsAg in CatL, probably due to the higher lymphatic targeting ability of NeuL. Thus, NeuL appears to be a better carrier for HBsAg than CatL. PMID- 10819419 TI - Intraliposomal chemical activation patterns of liposomal cis-bis-neodecanoato trans-R,R-1,2-diaminocyclohexane platinum (II) (L-NDDP)-a potential antitumour agent. AB - L-NDDP is a liposome-entrapped platinum compound currently in phase 2 clinical trials that has been shown to undergo intraliposomal activation. The degradation/activation kinetics of liposome entrapped cis-bis-neodecanoato-trans R,R-1,2-diamminocyclohexane platinum (II) [L-NDDP] at different conditions of pH, and temperature is presented. Liposomes were reconstituted in a solution of 0.9% sodium chloride (NaCl) in water (pH 5) at room temperature (formulation conditions currently used in the ongoing clinical trials). In the temperature experiments, L-NDDP 0.9% sodium chloride liposomes were incubated in a water-bath at 40, 60, and 80 degrees C. In the pH experiments, these solutions were compared to water, phosphate with and without chloride ion present, phosphate buffer without chloride ion at pH 3.1, 5.0, and 7.4, and glycine buffer with and without chloride ion. In 0.9% sodium chloride at room temperature, the chemical degradation/activation of liposome-bound NDDP was biphasic, with most of the degradation (approximately 45% conversion) occurring during the first hour after formation of the liposome suspension. NDDP degradation was pH dependent: when using pH 3 phosphate buffer as a reconstituting solution, liposome-bound NDDP degraded rapidly, whereas in pH 7.4 phosphate buffer it was stable for > 72 h. NDDP degradation was also temperature-dependent, the 50% point decreasing from 12 h at 25 degrees C to 9.5 h at 40 degrees C, 3.8 h at 60 degrees C, and 0.3 h at 80 degrees C when using 0.9% NaCl in water as a reconstituting solution. Using glycine buffer solution with and without NaCl at room temperature, no NDDP degradation over a 72 h period was observed at 25 degrees C; however, at 40 degrees C, only 68% NDDP remained intact at 72 h. Atomic absorption spectrophotometry (AAS) analysis of the eluting fractions after injection of L NDDP samples reconstituted in chloride-containing and non chloride-containing solutions clearly indicated that the formation of DACH-Pt-Cl2 was only observed when chloride-containing solutions were used and was first detected at 3 h when using 0.9% NaCl in water as a reconstituting solution. These results indicate that pH and temperature, and not the presence of chloride ion, are the main factors leading to the activation of NDDP. Since 45% of NDDP is already degraded at 1 h in the same conditions, it is concluded that (1) the first active intermediates of L-NDDP formed within the liposomes are the DACH-Pt chloro-aquo and diaquo intermediates, and (2) the in vivo, antitumour activity of L-NDDP is most likely mediated by direct intracellular delivery of the active species. PMID- 10819420 TI - A chemometrics method for separation of size dependent and size independent attributes of microspheres for medical ultrasound imaging. AB - The size dependent and size independent contributions to the mechanical properties of poly(ethyleneglycol) microspheres with encapsulated air, made for medical ultrasound imaging, have been studied. A relation between the size of microspheres in a reference group and their acoustic attenuation efficacy was derived by partial least square regression. The developed model was used to estimate acoustic attenuation as a function of the measured size distribution for various preparations made during formulation and process studies. The ratio between the measured attenuation and model estimated attenuation was then used to evaluate variations in the mechanical properties of the polymer shell and the contribution of such variations to the acoustical properties of the substance. The statistical parameter was compared with results for the mechanical compressibility derived from a theoretical approach and the results from the two methods mutually confirmed each other. The statistical procedure outlined has been found applicable for studies of other particulate products with size dependent in vitro or in vivo properties. PMID- 10819421 TI - Development of controlled release formulations of alachlor in ethylcellulose. AB - The herbicide alachlor (2-chloro-N-(2,6-diethylphenyl)-N-(methoxymethyl) acetamide) is frequently implicated in groundwater contamination. Microencapsulated alachlor should have reduced potential for leaching in the soil while maintaining effective biological activity. Microspheres of alachlor were prepared using ethylcellulose, according to the solvent evaporation method. The influence of formulation variables affecting the release rate of pesticide, such as the molecular weight of ethylcellulose, the amount of emulsifying agent, the pesticide/polymer ratio and the particle size, were investigated. The results showed that microspheres retarded the release of alachlor in different degrees. Pesticide/polymer ratio and particle size were the more important factors determining the alachlor release. Ethylcellulose microspheres may prove useful for the prolonged release of alachlor. PMID- 10819422 TI - Controlled delivery of drugs from a novel injectable in situ formed biodegradable PLGA microsphere system. AB - A novel method for in situ preparation of injectable biodegradable microspheres from the copolymer, poly(lactide-co-glycolide) (PLGA), without incorporating unacceptable organic solvents is described. The delivery system is a dispersion of PLGA microglobules ('premicrospheres' or 'embryonic microspheres') in an acceptable vehicle mixture (continuous phase) and whose integrity is maintained by the use of appropriate stabilizers. A solution of PLGA, triacetin, a model protein (cytochrome c), PEG 400, and Tween 80 (oil phase 1) is added dropwise with continuous homogenization to Miglyol 812-Span 80 solution (oil phase 2), thereby inducing phase separation (coacervation) of PLGA and forming PLGA microglobules (containing cytochrome c) dispersed in the continuous phase. This novel drug delivery system (NDDS) is a dispersion and has a viscous consistency, but is sufficiently syringeable. When injected, it comes in contact with water from an aqueous buffer or physiological fluid and, as a result, the microglobules harden to form solid matrix type microparticles entrapping cytochrome c (in situ formed microspheres). Cytochrome c is then released from these microspheres in a controlled fashion. The composition, rationale, and optimization of the NDDS are described here. Various formulation variables such as the PLGA concentration and type and the substitution of the continuous phase by a fresh oil phase 2 influenced the characteristics of this system. A preliminary investigation of the reproducibility and stability of the NDDS, as well as the physical stability of the encapsulated cytochrome c, revealed that these characteristics were not adversely affected. PMID- 10819423 TI - Effect of lecithins on BCG-alginate-PLL microcapsule particle size and stability upon storage. AB - Alginate-PLL microcapsules containing BCG were prepared by emulsification/internal gelation of an alginate solution dispersed within a vegetable oil containing lecithins as emulsifiers. The lecithins studied were soy bean lecithin at 0.1, 0.5, 1 and 2% concentration; and dried egg yolk lecithin at 0.1, 0.25, 0.5 and 1%. The microcapsule particle size showed a dependence upon the amount and type of lecithin added to the oil. Increasing the emulsifying agent concentration was found to reduce particle size, from 50.9 microm obtained when lecithins were not used in the emulsification step, to 13.9 microm obtained when 1% dried egg yolk lecithin was employed. The encapsulated BCG was identified by the Difco TB stain set K, followed by observation under optical microscopy. Once prepared, microcapsules were freeze-dried using 5% trehalose as cryoprotectant in order to preserve their stability upon storage. The stability of the microcapsules was assayed over 12 months at room temperature, finding that alginate-PLL microcapsules were stable up to 6 months. Moreover, in the case of microcapsules prepared with lecithins, a significant increase in particle size was observed, from 16.9 microm at the beginning of the study to 25.2 microm at 12 months storage. PMID- 10819425 TI - Literature alerts. PMID- 10819424 TI - In vitro release kinetics of Tolmetin from tabletted Eudragit microparticles. AB - In a previous paper the preparation has been described, by three different techniques, of microparticles made of Eudragit RS 100 and RL 100 containing a NSAI agent, Tolmetin. Freely flowing microparticles failed to affect significantly the in vitro drug release, which displayed a similar dissolution profile after micro-encapsulation to the free drug powder. Microparticles were then converted into tablets and the effect of compression on drug delivery, as well as that of the presence of co-additives, was studied in the present work. Furthermore, microparticles were also prepared by adding MgO to the polymer matrix, to reduce the sensitivity of the drug to pH changes during its dissolution. Similarly, magnesium stearate was also used for microparticle formation as a droplet stabilizer, in order to reduce particle size and hinder rapid drug release. A mathematical evaluation, by using two semi-empirical equations, was applied to evaluate the influence of dissolution and diffusion phenomena upon drug release from microparticle tablets. PMID- 10819427 TI - Assessment of attention following pre-school traumatic brain injury: a behavioural attention measure. AB - This study investigated attentional impairments and recovery in pre-school children following traumatic brain injury (TBI). Children aged between 3-8 years were assigned to two groups on the basis of severity of injury: (i) mild TBI and (ii) severe TBI. Children were evaluated at the acute stage postinjury (0-3 months) and again at 6 months on a range of standardized intellectual and attention measures. In addition, a specifically developed behavioural attention measure was employed. Results revealed a tendency for children with severe injuries to exhibit greater attentional impairments in the acute phase post injury. In addition, children who sustained a severe TBI demonstrated significant recovery in attentional functioning over time. Results indicated differential recovery rates for separate components of attention. Both groups demonstrated a trend towards recovery of arousal and motivation over time, while focused attention, impulsivity and hyperactivity remained stable. PMID- 10819426 TI - Early effects of traumatic brain injury on young children's language performance: a preliminary linguistic analysis. AB - Language skills undergo rapid development during the early childhood years, so that by the time children start school they are competent communicators with well established syntactic, semantic and pragmatic abilities for their age. Little is known about the effects of traumatic brain injury (TBI) on the acquisition of these language skills during the early childhood years. This study used a prospective, cross-sectional design to compare the language abilities of young children following their head injury. Fifteen brain injured children, aged between 4-6 years, were divided into three injury groups depending on severity of injury, i.e. mild, moderate and severe, and compared with a matched community control group. They were assessed within 3 months of sustaining their injury on a range of expressive and receptive language tests, and free speech conversation samples, which were analysed pragmatically and syntactically. Results indicated that the severe group performed most poorly on language tasks. It is suggested that linguistic evaluation is an important component of follow up at least for the severe head injured population. PMID- 10819428 TI - Functional memory skills following traumatic brain injury in young children. AB - Traumatic brain injury (TBI) may have a profound impact on a child's ongoing development. Various risk factors have been found to predict outcome, but considerable variability remains unexplained. This study used a prospective, longitudinal design to examine recovery of memory function following TBI within the pre-school period. Forty-four children with TBI were divided according to injury severity (mild, moderate, severe), and compared to age and SES matched healthy controls (n = 26). Children were evaluated acutely and at 12 months post injury using the Rivermead Behavioural Memory Test for Children. Results failed to show a clear dose-response relationship between injury severity and memory function during the acute phase of recovery. However, this relationship developed over time, with greater memory impairments evident for children with more severe TBI by 12 months post-injury. Children with mild TBI exhibited few memory problems. PMID- 10819429 TI - Recovery of educational skills following paediatric traumatic brain injury. AB - Academic success in the classroom is often dependent upon a child's ability in the areas of literacy, such as reading and spelling, and arithmetic. Following traumatic brain injury these skills are often compromised. The present study examined the recovery of educational skills (reading accuracy, reading comprehension, spelling and arithmetic) over 24 months post-injury, in a group of children who had sustained a mild, moderate or severe TBI. Results showed that the severe TBI group exhibited greater deficits on reading comprehension and arithmetic, while the moderate and severe TBI groups performed similarly in the areas of reading accuracy and spelling. Future research is required to further investigate predictors of educational outcome post-TBI. PMID- 10819430 TI - The development of pragmatic communication skills in head injured children. AB - This study investigated the developmental levels of pragmatic language skills in children following head injury (HI), in comparison to their uninjured peers. Participants were 30 head-injured and 19 healthy controls, classified into a 'young' age group, 8-9 years, and an 'old' age group, 11-12 years. Participants were administered the WISC- III, a negotiating requests task and a hint task, the latter two assessing verbal reasoning skills and abilities to be indirect, respectively. It was found that negotiation and hinting strategies were rapidly developing in these age groups, where abilities to hint were less mature for all groups. Results found a main effect for injury on cognitive and functional language tasks, reflected by lower performance levels and inflexibility in reasoning for the head-injured group. Injury sustained at an earlier age consistently predicted poorer performance on the language tasks, complicating the ongoing development of generalized and higher-order communicative skills. Severity of injury did not predict performance on either language task. PMID- 10819431 TI - Specific attention and executive function deficits in the long-term outcome of severe closed head injury. AB - Twenty-eight children (mean age= 10.9 years, SD= 1.9 years) who had experienced a severe closed head injury were assessed for outcome on tests of intelligence, attention and executive function at 1 year post-injury. The children with closed head injury were matched with children from a normative sample for age and intellectual ability. Specific deficits in attention and executive function were found in the closed head injury sample. The importance of including these outcome variables in child neuropsychological assessment is supported. PMID- 10819432 TI - Does family functioning affect outcome in children with neurological disorders? AB - Children with neurological disorders are at increased risk of poor psychological and social adjustment. Medical factors are insufficient to explain the individual variation in outcome, so family variables have been considered. This review evaluates studies which have investigated the effect of family functioning on outcome in children with traumatic brain injuries, epilepsy and other neurological disorders. Methodological problems include the cross-sectional nature of many studies and inadequate control for confounding variables. There is good evidence that family functioning influences behavioural adjustment and adaptive functioning after traumatic brain injury. The role of family functioning in epilepsy and other neurological disorders requires further investigation. The development of effective interventions to improve family functioning may lead to important clinical benefits for children and their families. PMID- 10819433 TI - Novel approaches to the treatment of hepatitis C virus infection. AB - Hepatitis C virus (HCV) has infected millions of people worldwide and emerged as a global health crisis. This review reports approaches currently being taken to combat the virus. Viral targets have received the most attention, particularly the NS3 serine protease where potent inhibitors have been described. Crystal structures of key replicative enzymes, NS3 protease, NS3 helicase, NS5B polymerase and now full-length NS3 protease-helicase, are available. More recently, targeting the host system has become of interest, particularly inhibitors of inosine monophosphate dehydrogenase. Research aimed at novel treatments for HCV disease is gathering pace and very recent developments in cell based assay systems can only hasten the discovery of improved therapies. PMID- 10819434 TI - Synthesis and anti-HIV activity of prodrugs derived from saquinavir and indinavir. AB - With a view to improving the pharmacological properties, safety and pharmacokinetic profiles of current protease inhibitors, the synthesis of various acyl-substituted saquinavir and indinavir prodrugs, their in vitro stability with respect to hydrolysis and their anti-HIV (LAI and HTLV IIIB) activity and cytotoxicity in CEM-SS and MT4 cells have been investigated. Hydrolysis of the ester bond and liberation of the active free drug was found to be crucial for HIV inhibition: the faster the hydrolysis, the closer the anti-HIV activity was to that of the respective parent drug. This is the case for most of the C-14 substituted indinavir and saquinavir derivatives (IC50 from 10 to 360 nM for ester half-lives of 90 min to 40 h). Concomitantly, the level of HIV inhibition is very low for the prodrugs for which hydrolysis is very slow. This is the case with the myristoyl or oleyl saquinavir esters, owing to the stable masking of the hydroxyl that is part of the peptidomimetic non-cleavable transition state isostere responsible for the inhibitory potency of saquinavir (and indinavir). In contrast, the anti-HIV activity of the monosubstituted C-8 indinavir prodrugs seems not to be correlated with their resistance to hydrolysis, as expected (the C-8 hydroxyl of indinavir is not involved in the transition state isostere). No cytotoxicity was detected for the indinavir and saquinavir prodrugs for concentrations as high as 10 or even 100 microM, thus indicating promising therapeutic potential. PMID- 10819435 TI - Phosphoramidate derivatives of stavudine as inhibitors of HIV: unnatural amino acids may substitute for alanine. AB - Some novel phosphoramidate derivatives of the nucleoside analogue stavudine have been prepared as membrane-soluble prodrugs of the bioactive free phosphate forms. Phenyl phosphates linked via nitrogen to methyl esterified amino acid analogues were studied, where the amino acid was an unnatural alpha-alkyl (or aryl) glycine or an alpha,alpha-dialkyl glycine. All compounds were characterized by a range of spectroscopic, spectrometric and analytical methods and were subjected to in vitro evaluation of their anti-human immunodeficiency virus efficacy. It is notable that certain unnatural amino acid derivatives could substitute for alanine with only a relatively small loss of activity and, moreover, that this activity did not fall-off with increasing alkyl chain length for the C2-C4 mono alkyl series. These data are further probed by the application of our recently reported 31P-NMR-based carboxyl esterase assay, with informative results. PMID- 10819436 TI - Structure-activity relationship studies on potential non-nucleoside DABO-like inhibitors of HIV-1 reverse transcriptase. AB - Using 2,6-dichloro-4-aminopyrimidine, a number of uracil and cytosine derivatives with both arylthio and alkoxy moieties were prepared. These novel pyrimidines share chemical similarities with DABOs and HEPTs, two classes of non-nucleoside human immunodeficiency virus type 1 (HIV-1) reverse transcriptase inhibitors (NNRTIs), which have been widely studied of late. All new derivatives were tested in MT-4 cells to explore their potential in vivo anti-HIV activity. Like other NNRTIs, they selectively inhibit HIV-1 but not HIV-2. The majority of test derivatives were found to have low potency and were sometimes more cytotoxic than zidovudine and emivirine (formerly MKC-442), used here as reference drugs. Uracil and cytosine derivatives bearing a sec-butoxy chain and a methyl-substituted benzenesulphonyl moiety were the most potent. Enzyme assays proved that these derivatives target RT. Structure-activity relationship studies established a correlation between the anti-HIV-1 activity and the meta substitution on the phenyl ring; furthermore, oxidation of sulphide to sulphone significantly increased the potency of certain derivatives. PMID- 10819437 TI - N-[2-(4-methylphenyl)ethyl]-N'-[2-(5-bromopyridyl)]-thiourea as a potent inhibitor of NNRTI-resistant and multidrug-resistant human immunodeficiency virus type 1. AB - The composite non-nucleoside reverse transcriptase inhibitor (NNRTI) binding pocket model was used to study a number of thiourea analogues with different substitutions at the 4-phenyl position including N-[2-(4-methylphenyl)ethyl]-N' [2-(5-bromopyridyl)]-thiourea (compound HI-244), which inhibited recombinant RT better than trovirdine or compound HI-275 with an unsubstituted phenyl ring. HI 244 effectively inhibited the replication of HIV-1 strain HTLV(IIIB) in human peripheral blood mononuclear cells with an IC50 value of 0.007 microM, which is equal to the IC50 value of trovirdine. Notably, HI-244 was 20 times more effective than trovirdine against the multidrug-resistant HIV-1 strain RT-MDR with a V106A mutation (as well as additional mutations involving the RT residues 74V, 41L and 215Y) and seven times more potent than trovirdine against the NNRTI resistant HIV-1 strain A17 with a Y181C mutation. PMID- 10819438 TI - Non-nucleoside HIV-1 reverse transcriptase inhibitors: synthesis and biological evaluation of novel quinoxalinylethylpyridylthioureas as potent antiviral agents. AB - New heterocyclic derivatives of ethylpyridylthiourea, quinoxalinylethylpyridylthiourea (QXPT) and analogues, inhibited human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) activity and prevented HIV-1 cytopathogenicity in T4 lymphocytes. Several of these novel non nucleoside RT inhibitors, with a substituted pyrroloquinoxalinone heteroaromatic skeleton, showed inhibitory activity against wild-type RT as well as against mutant RTs containing the single amino acid substitutions L1001, K103N, V106A, Y1811 and Y188L that was much greater than other non-nucleoside inhibitors such as nevirapine. Maximum potency in enzymatic assays was achieved with a fluoropyrroloquinoxaline skeleton linked to the ethylpyridylthiourea moiety (FQXPT). In cell-based assays on different cell lines and on human monocyte macrophages, 6-FQXPT exhibited EC50 values in the nanomolar range, with a promising selectivity index. Moreover, 6-FQXPT showed synergistic antiviral activity with zidovudine. PMID- 10819439 TI - Extract of Prunella vulgaris spikes inhibits HIV replication at reverse transcription in vitro and can be absorbed from intestine in vivo. AB - It has been reported that extracts of the spike of Prunella vulgaris (PS) exhibit anti-HIV activity at the adsorption and reverse transcription stages. In this study, the actual activity of PS in cells, kinetic analysis of the inhibitory activity of PS against HIV reverse transcriptase and the feasibility of oral administration were examined. First, to clarify whether this extract shows anti HIV activity in cells in vitro, the number of copies of proviral DNA in HIV exposed cells was calculated. The number of copies was significantly decreased in cells cultured in the presence of PS extract, but not in the presence of dextran sulphate. The activity of PS extract in the cells was also assessed by the drug addition test, during and after HIV adsorption. PS extract and dextran sulphate suppressed HIV production to similar levels when added after HIV adsorption. However, only PS extract suppressed HIV production at the same concentration when the drugs were added during HIV adsorption. Presumably, the penetration of the PS extract into the cells was required for this activity. Secondly, fractionated PS inhibited HIV reverse transcription in a non-competitive manner. This fractionated PS kept anti-HIV activity, but inhibited HIV replication and adsorption to a lesser extent compared to dextran sulphate. Lastly, an active component(s) was detected in plasma in vivo, after injection into the intestine, which demonstrates the feasibility of oral administration dosing. PMID- 10819440 TI - Peyronie's disease: etiology, medical, and surgical therapy. AB - Peyronie's disease remains an enigma. With the recent introduction of an animal model for Peyronie's disease, the entry of a number of double-blind placebo controlled clinical trials, and the application of new molecular diagnostic methods, the investigation of this wound-healing disorder of the penile tunica albuginea should illuminate many of the unknowns. Investigators need to be open to innovations in other fields of medicine involving idiopathic fibrosing conditions in other organ systems, eg, Dupuytren's contracture, keloids, hypertrophic scarring, etc. Applications from these other disciplines will undoubtedly widen our scope about Peyronie's disease. While a minority of patients respond with observation alone, most authorities recommend at least a trial of medical therapy with a safe, inexpensive, and well-tolerated agent, as early-stage disease is reputedly more likely to respond better than patients with established, longstanding Peyronie's plaques. The reintroduction of intralesional therapies (verapamil and interferon alpha-2b) provides the clinician with an alternative minimally invasive intervention that has promising possibilities. In severe fibrotic or calcified plaques or with major structural abnormalities, the judicious use of surgery with or without grafting materials and a penile prosthesis can restore many men back to their previous level of high esteem and provide both partners an excellent quality of life. PMID- 10819441 TI - Fertilizing ability of epididymal sperm from dead animals. PMID- 10819442 TI - Genetic fixes and future generations. PMID- 10819443 TI - Sperm preparation methods. PMID- 10819444 TI - Bombesin modulates the association of Src with a nuclear 110-kd protein expressed in dividing prostate cells. AB - Polyclonal antibodies produced against a peptide derived from the Fer tyrosine kinase sequence also specifically recognized a 110-kd protein (p110) up-regulated in dividing versus resting dog prostate epithelial cells in vitro. In vivo in the dog prostate, p110 expression was detected when basal cell metaplasia was induced by estrogens after castration but not when renewing the differentiated epithelium with androgens. It was also detected in extracts from the human prostatic carcinoma cell lines LNCaP, DU145, and PC-3, and from 6 out of 11 human prostate cancer tissues analyzed, but not from normal or hyperplastic glands. The tyrosine kinase Src was shown by coimmunoprecipitation to associate with p110, and this interaction was positively modulated by bombesin stimulation of PC-3 cells. However, p110 was not tyrosine phosphorylated. Moreover, it was mainly distributed in the nuclear fraction. This nuclear p110 protein, expressed in dividing prostate epithelial cells and in human prostate cancer cells and tissues, could thus be a downstream mediator of bombesin-signaling pathways, acting via its association with Src. PMID- 10819445 TI - Immunolocalization of CA II and H+ V-ATPase in epithelial cells of the mouse and rat epididymis. AB - Acidification of the epididymal lumen has been suggested to play an important role in sperm functions; however, the cell types, pumps, and mechanisms involved have not been fully addressed. In this study, carbonic anhydrase II (CA II) and a 67-kd subunit of Neurospora crassa vacuolar proton adenosinetriphosphatase (H+ V ATPase) pump were immunolocalized using light microscopy and electron microscopy (EM) in the epididymis of rats and mice. In both animals, narrow cells, identified in the initial segment and intermediate zone of the epididymis, contained numerous small vesicles in their apical region, often cup-shaped in appearance. In the mouse but not rat, these cells also possessed numerous cisternae of smooth endoplasmic reticulum, suggesting steroid synthesis; and cytoplasmic blebs of their apical cell surface, which appeared to detach, suggesting apocrine secretion. Anti-CA II antibody was immunocytochemically localized in the light microscope within narrow cells but not over any other cell types of the entire epididymis. Anti-H+ V-ATPase antibody was also localized in narrow cells of the initial segment and intermediate zone; as well as clear cells of the caput, corpus, and cauda regions. Using EM, gold particles for anti-CA II and H+ V-ATPase antibodies were noted in the apical region of narrow cells in relation to the numerous, small, cup-shaped vesicles. Although CA II was mainly located in the cytosol near these vesicles, H+ V-ATPase appeared on their delimiting membrane and on the apical plasma membrane of these cells. A similar distribution was noted for H+ V-ATPase in clear cells. The nature of the small vesicles of the apical region of narrow cells was examined with electron-dense fluid phase tracers that were introduced into the epididymal lumen. The tracers appeared within these vesicles and a few endosomes 1 hour after injection, suggesting that they contact the apical plasma membrane. Since these vesicles are also related to CA II and H+ V-ATPase, the data suggests that, as the site of proton production, the vesicles recycle to and from the apical cell surface, and in this way, deliver protons to the epididymal lumen for acidification. Clear cells and their expression of H+ V-ATPase may also serve in this function. In summary, both narrow and clear cells appear to be involved in luminal acidification, an activity that may be essential for sperm as they traverse and are stored in the epididymis. PMID- 10819446 TI - Effects of luteinizing hormone, follicle-stimulating hormone, and epidermal growth factor on expression and kinase activity of cyclin-dependent kinase 5 in Leydig TM3 and Sertoli TM4 cell lines. AB - We examined the effects of luteinizing hormone (LH), follicle-stimulating hormone (FSH), and epidermal growth factor (EGF) on the expression and kinase activity of cyclin-dependent kinase 5 (Cdk5) in Leydig TM3 and Sertoli TM4 cell lines. Hormonal regulation of the expression and activity of Cdk5 by using normal and hypophysectomized rat testes was also investigated to elucidate its role. Cdk5 levels and kinase activity were significantly elevated in TM3 cells that were grown in the presence of 7.5% serum, EGF, or LH and were associated with an increase in testosterone production compared with controls. These increases were accompanied by an increase in proliferation of TM3 cells after treatment with serum or EGF but not with LH suggest that Cdk5 may be involved in cellular differentiation that is induced with LH treatment. In contrast, the presence of neither serum, EGF, nor FSH had a significant effect on Cdk5 activity levels in the Sertoli TM4 cell line, and there was no correlation with proliferative activity or transferrin levels. A significant decrease in Cdk5 expression and activity were noted in rat testis after hypophysectomy compared with normal rat testis and is associated with a simultaneous decrease in testosterone and transferrin levels. Immunohistochemical analysis revealed that Cdk5 was strongly expressed in the nuclei and cytoplasm of Leydig cells, Sertoli cells, spermatogonia, and peritubular cells of normal adult rat testis. After hypophysectomy, the pattern of Cdk5 staining differed markedly from that in normal rat testis and a profound reduction in staining of Cdk5 was observed in each tubule. Our results suggest that LH and EGF influence and modulate Cdk5 expression and activity in Leydig TM3 cells and may, conceivably, be involved in signal transduction cascades that are initiated by hormones or growth factors. Cdk5 in Sertoli TM4 cells is likely to possess some constitutive functions that are not affected by the cells' proliferation state. Moreover, Cdk5 is probably involved in the constitutive and hormonally stimulated activities of the rat testis, in addition to its involvement in cell proliferation. PMID- 10819447 TI - Accidental radiation exposure and azoospermia. AB - Seven Georgian male soldiers (19-25 years old) had accidentally been exposed to radiation by Cs-137 between April 1996 and May 1997. No information about the exact time and duration of exposure was available. All patients presented with the subacute stage of Cutaneous Radiation Syndrome with deep painful ulcers on different body sites, predominantly on the legs. Semen analyses showed complete azoospermia in 4 patients, with elevated follicle-stimulating hormone (FSH) in 3 and elevated luteinizing hormone (LH) in 2 of them. One patient had severe oligozoospermia of 7 million sperm per mL, with normal sperm motility and morphology; his FSH and LH levels were elevated. One patient had complete normozoospermia, and the seventh patient had polyzoospermia of 340 million per mL; both of these patients had normal serum hormone levels. Only the patient with oligozoospermia reported a history of delayed testicular descent; his physical examination showed relatively soft and small testicles and a varicocele with considerable reflux. The physical andrological examinations were normal in the other 6 patients. It is very likely that the azoospermia in the 4 patients can be attributed to the radiation accident. In conclusion, it is essential to perform andrological examinations in patients who have been exposed to radiation even if there are only cutaneous injuries detectable, as a high percentage of them can show azoospermia. PMID- 10819448 TI - Cryopreservation of a small number of fresh human testicular spermatozoa and testicular spermatozoa cultured in vitro for 3 days in an empty zona pellucida. AB - It is known that the motility of human testicular sperm can be improved when they are cultured in vitro for a few days. The purpose of this study was to determine whether it is better to freeze human testicular spermatozoa on the day of biopsy (fresh) or after they were cultured for 3 days. A modified, single-sperm freezing technique was used in this study. The study consisted of two parts: (1) ejaculated spermatozoa were used to examine the influence of different concentrations of glycerol and synthetic serum substitute (SSS) on the survival rate after cryopreservation, and (2) the survival rates between cryopreserved fresh testicular spermatozoa (Group 1) and testicular spermatozoa that were cultured for 3 days before freezing (Group 2) were compared. Empty zonae pellucidae were obtained from mouse eggs. Five to 10 motile spermatozoa were selected and injected into an empty zona pellucida. For freezing, the zona pellucida with spermatozoa was transferred into a HEPES-buffered human tubal fluid containing different concentrations of glycerol and kept at room temperature for 10 to 15 minutes, and then loaded into a 0.25-ml-plastic straw. The straws were exposed to liquid nitrogen vapor for 2 hours and then plunged into liquid nitrogen. For thawing, the straws were taken out of liquid nitrogen and placed into a 37 degrees C waterbath for 25 to 30 seconds. There was no statistically significant difference in survival rates between 3% and 10% SSS with different glycerol concentrations. There was no statistically significant difference in the survival rates of spermatozoa between Group 1 and Group 2 after cryopreservation. It appears that in vitro culture of testicular spermatozoa before freezing does not increase survival rate. PMID- 10819449 TI - Osteopontin gene expression in the Holstein bull reproductive tract. AB - The objective of this study was to localize gene expression of osteopontin in the Holstein bull reproductive tract using Northern blot analysis and in situ hybridization. For Northern blot analysis, a digoxigenin-labeled osteopontin complementary deoxyribonucleic acid (cDNA) was used to probe blots containing total ribonucleic acid (RNA) isolated from the testis, epididymis, vas deferens, ampulla, seminal vesicle, prostate, and bulbourethral glands. The digoxigenin labeled cDNA for the bovine homologue of osteopontin was hybridized to a single band at approximately 1.6 kb to RNA samples from the ampulla and seminal vesicle. For in situ hybridization studies, antisense and sense riboprobes were synthesized and used to hybridize cryosections that had been obtained from bull reproductive tissues. In situ hybridization of the bull testis detected osteopontin messenger RNA in the developing germ cells. Osteopontin gene expression was detected only in seminiferous tubules that contained elongated spermatids, which suggests that expression varies with the stage of the seminiferous epithelium. Within the epididymis, silver grains were distributed over the sperm that were located within the lumen of the caput, corpus, and cauda epididymis. Osteopontin expression was primarily observed in the epithelial cells of the ampulla. Antisense riboprobes also hybridized to sperm that were located within the lumen of the ampulla, confirming the presence of osteopontin transcripts in the haploid male gamete. PMID- 10819450 TI - Multiple promoter and splicing mRNA variants of the epididymis-specific gene EP2. AB - The EP2 gene codes for a family of androgen-dependent, epididymis-specific secretory proteins. Using probes derived from human HE2 cDNA, a chimpanzee epididymal cDNA library was screened. Five variants of chimpanzee EP2 cDNA were identified. Variant 1 (EP2A) is the chimpanzee ortholog of HE2. Variant 2 (EP2B) has an alternative 5' end. Variant 3 (EP2C) has an alternative 3' end. Two additional variants were identified by reverse transcriptase-polymerase chain reaction analysis. Variant 4 (EP2D) and variant 5 (EP2E) appear to lack an exon, resulting in a shift in the open reading frame. Presumably, the 5 variants originate from the same gene and result from alternative promoters and alternative splicing. Each of the putative proteins encoded by these variant messages has a leader sequence characteristic for a secretory protein. After removal of the leader sequence, each of these proteins is predicted to consist of 1 or 2 out of 4 possible peptide modules. Two of these modules have no recognizable homology to known proteins. The other 2 modules have a distribution of cysteine residues characteristic for beta-defensins, a family of proteins with antimicrobial activity. PMID- 10819451 TI - Posttesticular antifertility action of triptolide in the male rat: evidence for severe impairment of cauda epididymal sperm ultrastructure. AB - A variety of active diterpene epoxides, including the triptolide (isolated from Tripterygium wilfordii) have been reported to cause infertility in male rats. Previously, we showed that oral administration of triptolide at a dosage of 100 microg/kg per body weight for 70 days completely inhibited fertility in male rats, with little or no demonstrable detrimental effect on spermatogenesis and Leydig cell function as determined by testicular light microscopic appearance and serum and intratesticular testosterone levels. Despite the apparent absence of effects on the testes, cauda epididymal sperm were abnormal, with complete cessation of sperm motility and some reduction in sperm numbers. This study was undertaken to provide additional insight into the subcellular sites and possible mechanisms of action of this compound using ultrastructural analysis of the testes and epididymidis. The most striking effect of triptolide treatment was observed in sperm in the epididymis. In rats rendered infertile with 100 microg/kg per body weight of triptolide daily for 70 days, virtually all cauda epididymal sperm exhibited complete absence of plasma membrane over the entire middle and principal piece, premature decondensation of the nuclei, and disorganization of the mitochondrial sheath with many vacuolated mitochondria. No ultrastructural differences in the epididymal epithelium were observed between control and triptolide-treated rats. The testes appeared to be mildly affected after triptolide treatment but exhibited only subtle ultrastructural defects in the germ cells. The findings of severe impairment of cauda epididymal sperm ultrastructure, along with minimal discernible abnormalities in the fine structural cytology of the testes, further suggest that the site of action of this compound is posttesticular and may be confined to the cauda epididymal sperm. However, we cannot rule out an effect of triptolide that occurs during germ cell maturation but is delayed in its manifestation or triggered at the rete testis and epididymal level. PMID- 10819452 TI - Experimental cryptorchidism inhibited growth of the rat ventral prostate. AB - Adult Sprague-Dawley male rats, weighing about 350 g, were rendered cryptorchid by suturing the testes to the lateral abdominal wall. Twenty-eight days later, cryptorchidism resulted in a significant decline in testis weight and suppressed spermatogenesis. The ventral prostate was significantly smaller in cryptorchid rats. There was no significant difference in serum testosterone levels between the normal and cryptorchid rats. Charcoal-stripped aqueous extracts of the testis from intact and cryptorchid animals were tested on primary cultures of rat prostatic stromal cells. Cultures treated with extract from the intact testis had a significantly increased cell proliferation as assessed by cell count and by the rate of 3H-thymidine incorporation. Additionally, extracts of seminiferous tubules significantly increased prostate stromal cell proliferation compared to extracts of testicular interstitial components. Furthermore, this proliferative effect of testicular extracts is specific to the prostate as extract of both normal and cryptorchid testis stimulated proliferation of rat footsole fibroblasts in culture, but only extracts from intact testis stimulated proliferation of prostate stromal cells. These observations demonstrate that the testis produces nonandrogenic substances that can promote growth of prostatic stromal cells and that these substances were eliminated in the cryptorchid testis. PMID- 10819453 TI - The permeability of the microvasculature of the perfused rat testis to small hydrophilic substances. AB - The permeability-surface area products (PS) for sodium, Cr-EDTA, and cyanocobalamine (CoB12) have been determined in isolated perfused rat testes, using the single-passage multiple tracer technique, with albumin as the reference tracer. The validity of using albumin was established from its recovery in the perfusate leaving the testis, which was 98.73+/-0.48% of that for Cr-labeled red cells. The PS values obtained for Na, Cr-EDTA, and CoB12 were correlated with perfusate flow, both below and above levels that were equivalent to normal rates of blood flow in the testis (0.3 mL/[g x min]). The values found at the highest flow rates obtained (between 2.7 and 3.5 mL/[g x min]) were 2230+/-240 microL/(g x min) (n = 8) for sodium, 1460+/-140 microL/(g x min) (n = 7) for Cr-EDTA, and 850+/-80 microL/(g x min) (n = 7) for CoB12. These values are similar to those reported at equivalent flow rates for heart muscle and greater than those reported for skeletal muscle, both of which have unfenestrated capillaries similar to testis, but are less than the values for pancreas and salivary gland, which have fenestrated capillaries and are similar to most other endocrine tissues. However, the permeability coefficients for these markers in the testis (calculated using published values for the surface area of the testicular microvasculature) appear to be considerably greater than for any other tissue studied thus far. By extrapolating extraction values, either linearly or logarithmically, to obtain maximal values for PS for Cr-EDTA and CoB12, and comparing the ratio of these PS area values with the ratio of the diffusion coefficients of these molecules, it can be calculated that the equivalent pore radius for the testicular endothelium is between 5 and 6 nm, comparable to those calculated for other nonfenestrated endothelia. PMID- 10819454 TI - The uptake of amino acids, in particular leucine, by isolated perfused testes of rats. AB - The uptake of amino acids by the isolated rat testis perfused with Krebs-Ringer solution with albumin has been studied using the single-passage, multiple-tracer technique with [14C]-mannitol as the reference tracer. When the perfusate contained no added amino acids, the uptake of [3H]-Leu was between 60% and 80% of the uptake of mannitol at all times after injection of the bolus; there was a small but significant uptake of some other amino acids studied (Ala, Gly, Glu, and Asp); and with Ala, Glu, and Asp, uptake increased slightly in increasing times after injection. There was no significant uptake of Arg. The uptake of Leu could be decreased by the inclusion of nonradioactive Leu in the perfusate, and the Km and Vmax of the transport were 0.067 mM and 19.5 nmole/(g x min), respectively. The Km value is similar to that for transport into brain and much less than the values obtained in other tissues for the related amino acid Phe, which is transported by the same L system. The transport of L-Leu in the testis was also inhibited by L-Phe or D-Phe, D-Leu, and by the synthetic amino acid Bch, the characteristic marker for the L system, but was only slightly reduced if the perfusate was free of sodium, as is expected for the L system. By autoradiography after fixation by perfusing with glutaraldehyde, the transport of Leu could be localized to the endothelial cells of the larger vessels of the testicular microvasculature. PMID- 10819455 TI - Mechanism of protection of rat spermatogenesis by hormonal pretreatment: stimulation of spermatogonial differentiation after irradiation. AB - Pretreatment of rats with hormones that suppress testosterone levels and sperm production enhances the recovery of spermatogenesis from stem cells after a cytotoxic insult. It is not known whether the enhanced recovery results from an increase in the numbers of surviving stem cells or whether their ability to differentiate is enhanced. In this study, untreated rats and rats pretreated with testosterone plus estradiol-17beta (T + E) were irradiated with 3.5 or 6 Gy, and the recovery of spermatogenesis from surviving stem cells was assessed at 6, 10, and 20 weeks after irradiation. T + E pretreatment did not significantly affect the numbers of A spermatogonia remaining in the tubules at 6 weeks after irradiation. In rats that were given irradiation alone, spermatogenesis steadily declined after 6 weeks because the stem cells lost their ability to differentiate. However, when rats were treated with T + E before irradiation, this decline was prevented, and in fact, at least at the lower dose of radiation, there was a progressive recovery of spermatogenesis. Given the similar spermatogonial counts at 6 weeks after irradiation in the irradiated-only and T + E-treated, irradiated rats, the hormone treatment appears not to protect stem cells from being killed by the cytotoxic agent. Rather, the later enhancement of spermatogenic recovery results from prevention of an injury-induced change in spermatogonia or in their environment, which would have otherwise resulted in failure of spermatogonial differentiation. PMID- 10819456 TI - Dynamic subcellular distribution of the DAZL protein is confined to primate male germ cells. AB - We have examined the location of the DAZL protein in fetal and adult rodents and human specimens and found that there is a shift from a predominantly nuclear to a predominantly cytoplasmic distribution of the protein in human testis. In rat testis and human ovary, however, the protein is predominantly, if not exclusively, cytoplasmic throughout germ cell development. One possible explanation for this could be that the DAZ protein is responsible for the nuclear localization of DAZL in human males. We have tested this hypothesis by examining the testis of marmosets, which lack the Daz genes and have found that the DAZL protein is both nuclear and cytoplasmic in spermatogonia, and by analyzing testis sections from DAZ-deleted patients in whom the cytoplasmic location of DAZL is evident in remaining germ cells. Transfection experiments indicate that the differences in DAZL expression between rodents and humans are not caused by the amino acid differences between the 2 proteins, and that DAZL is a cytoplasmic protein per se. Variations in location seem to be independent of the presence of the DAZ protein are species specific and, as in Drosophila, may not have great functional significance. PMID- 10819458 TI - As we celebrate the 25th anniversary of our Society, it is important to briefly review the events that led to its beginning. PMID- 10819457 TI - Evaluation of a container for collection and shipment of semen with potential uses in population-based, clinical, and occupational settings. AB - Large, population-based studies of semen quality are encumbered by the logistics and expense of obtaining semen samples from men who live in a variety of locations. A prototype semen collection and transportation kit, the TRANSEM100, can be distributed to study participants and then directly shipped to a central laboratory for analysis. This study was designed to evaluate the ability of male volunteers to correctly use the kit. Thirty volunteers aged 20 to 44 years with no history of diabetes, recent chemotherapy, fertility problems, or vasectomy were recruited through a newspaper advertisement, interviewed to obtain demographic information, and instructed on the use of the kit. Twenty-six of the initial subjects provided at least 1 semen specimen using the kit and returned the specimens by overnight delivery to the laboratory for analysis, 25 completed a follow-up interview on the use of the collection kit, and 20 submitted a second semen sample using the same method. The average volunteer was white, 27.8 years old, and held at least a college degree. Forty percent of the volunteers were married. In general, participants correctly followed the instructions for collecting, packaging, and shipping the semen samples. Volunteers were instructed to collect samples after at least 2, but no more than 7 days of abstinence. For the first and second samples submitted, participants collected semen samples after an average of 3.3 and 3.9 days of abstinence, respectively. Seventeen (65%) of the samples from the first sampling period and 16 (80%) of the samples from the second period were received in the laboratory the day after they had been collected. In summary, the TRANSEM100 may prove to be useful for collecting human semen in field studies. Further testing of this method is warranted to evaluate preservation of sample quality and use of the kit by men among diverse socioeconomic groups. PMID- 10819459 TI - Pretreatment platelet 5-HT concentration predicts the short-term response to paroxetine in major depression. PMID- 10819460 TI - Why zinc in zinc enzymes? From biological roles to DNA base-selective recognition. AB - Intrinsic chemical properties of the zinc(II) ion in zinc enzymes have been investigated by the model of 1:1 Zn2+-macrocyclic polyamine complexes, including Zn2+-1,5,9-triazacyclododecane ([12]aneN3) and 1,4,7,10-tetraazacyclododecane (cyclen). The physiologically most suitable pKa values for the Zn2+-bound H2O in enzymes were illustrated by the first model Zn2+-[12]aneN3 complex, which mimics the essential kinetic and thermodynamic roles of Zn2+ in carbonic anhydrase. The activation of proximate serine residues (in alkaline phosphatase) and activation of alcohols for hydride transfer to NAD+ (in alcohol dehydrogenase) were also mimicked by Zn2+ -[12]aneN3 complexes. The functions of two zincs in dinuclear metallophosphatases were explained by a new dinuclear Zn2+-cryptate. For an aldolase type II model, a Zn2+-cyclen derivative showed facile enolate formation from a proximate carbonyl pendant under physiological conditions. The strong anion affinities, which Zn2+ intrinsically possesses, were exploited into novel selective nucleobase thymine (or uracil) recognition of Zn2+-cyclen complexes by the strong Zn2+ -imido anion bond formation. The Zn2+-aromatic-pendant cyclen complexes selectively bind to T (or U) in single- and double-stranded DNA (or RNA). Thus, Zn2+ complexes act like molecular zippers to break A-T pairs in DNA, which was proven by various physicochemical measurements and DNA footprinting assays. These Zn2+ complexes showed some relevant biochemical and biological properties such as inhibition of transcriptional factor, TATA binding protein, or strong antimicrobial activities to gram-positive bacterial strains. PMID- 10819461 TI - In situ X-ray absorption spectroelectrochemical study of hydroxocobalamin. AB - An in situ X-ray absorption spectroscopy (XAS) spectroelectrochemical study of aquocobalamin (system B12a-B12r-B12s) has been carried out in aqueous solutions buffered at different pH values. To the best of our knowledge, this is the first structural study of aquocobalamin at room temperature under controlled oxidation conditions. Most of the previous work was in fact performed using frozen samples chemically treated to produce the species. The spectroelectrochemical approach offers several advantages: (1) the reduction products may be studied without poisoning the system with chemical reductive reagents and (2) any possible variation of the oxidation state owing to the electrons produced by the incident beam is avoided as the electrode, under potentiostatic control, acts as a scavenger. The spectroelectrochemical approach, together with more careful data analysis, has led to an improved interpretation of the XAS data. These conditions were not met in previous works where the oxidation state was not controlled and multiple scattering contributions were not taken into account. The general shape of the XAS spectra of the different species is not greatly affected by pH. A signature for the base-off square-planar coordination has been evidenced for the Co(II) compound at basic pH. A new signature for Co(I), indicating square-planar coordination, has been identified on the experimental spectra and simulated in theoretical X-ray absorption near-edge structure (XANES) studies. The flexibility of the electrochemical approach, that permits to unambiguously establish the formal oxidation state, has led to very reliable values for energy shift and peak intensity variations. The experimental XANES and extended X-ray absorption fine structure (EXAFS) spectra with a very good signal-to-noise ratio have been processed using the GNXAS package that takes into account multiple scattering contributions. EXAFS and XANES independent analysis result in the same structural model. The reduction from Co(III) to Co(II) produces the most significant structural changes: the cobalt coordination number decreases from six to five, and the edge position shifts by 2.4 +/- 0.3 eV. In addition, the XANES spectra are strongly modified. The reduction from Co(II) to Co(I) produces mainly electronic effects with no apparent change of the coordination number. A discussion of the limits and potentialities of EXAFS in this type of study has also been included. PMID- 10819462 TI - Modular organization and identification of a mononuclear iron-binding site within the NifU protein. AB - The NifS and NifU nitrogen fixation-specific gene products are required for the full activation of both the Fe-protein and MoFe-protein of nitrogenase from Azotobacter vinelandii. Because the two nitrogenase component proteins both require the assembly of [Fe-S]-containing clusters for their activation, it has been suggested that NifS and NifU could have complementary functions in the mobilization of sulfur and iron necessary for nitrogenase-specific [Fe-S] cluster assembly. The NifS protein has been shown to have cysteine desulfurase activity and can be used to supply sulfide for the in vitro catalytic formation of [Fe-S] clusters. The NifU protein was previously purified and shown to be a homodimer with a [2Fe-2S] cluster in each subunit. In the present work, primary sequence comparisons, amino acid substitution experiments, and optical and resonance Raman spectroscopic characterization of recombinantly produced NifU and NifU fragments are used to show that NifU has a modular structure. One module is contained in approximately the N-terminal third of NifU and is shown to provide a labile rubredoxin-like ferric-binding site. Cysteine residues Cys35, Cys62, and Cys106 are necessary for binding iron in the rubredoxin-like mode and visible extinction coefficients indicate that up to one ferric ion can be bound per NifU monomer. The second module is contained in approximately the C-terminal half of NifU and provides the [2Fe-2S] cluster-binding site. Cysteine residues Cys137, Cys139, Cys172, and Cys175 provide ligands to the [2Fe-2S] cluster. The cysteines involved in ligating the mononuclear Fe in the rubredoxin-like site and those that provide the [2Fe-2S] cluster ligands are all required for the full physiological function of NifU. The only two other cysteines contained within NifU, Cys272 and Cys275, are not necessary for iron binding at either site, nor are they required for the full physiological function of NifU. The results provide the basis for a model where iron bound in labile rubredoxin-like sites within NifU is used for [Fe-S] cluster formation. The [2Fe-2S] clusters contained within NifU are proposed to have a redox function involving the release of Fe from bacterioferritin and/or the release of Fe or an [Fe-S] cluster precursor from the rubredoxin-like binding site. PMID- 10819463 TI - The interactions of square platinum(II) complexes with guanine and adenine: a quantum-chemical ab initio study of metalated tautomeric forms. AB - The influence of binding of square planar platinum complexes on tautomeric equilibria of the DNA bases guanine and adenine was investigated using the density functional B3LYP method. Neutral trans-dichloro(amine)-, +1 charged chloro(diamine)-, and +2 charged triamine-platinum(II) species were chosen for coordination to bases. Only the N7 interaction site of the bases was considered. The calculations demonstrate that the neutral platinum adduct does not change the tautomeric equilibria of the bases. Furthermore, N7 binding of the neutral Pt adduct moderately reduces the probability of protonation of the N1 position of adenine. Larger effects can be observed for +1 and mainly +2 adducts, but these can be rationalized by electrostatic effects. Since the electrostatic effects are expected to be efficiently compensated for by a charged backbone of DNA and counterions in a polar solvent, no dramatic increase in mispair formation is predicted for Pt(II) adducts, which is in agreement with experiment. The interaction energies between Pt adducts and the nucleobases were also evaluated. These interaction energies range from ca. 210 kJ/mol for neutral adducts, interacting with both bases and their tautomers, up to 500 kJ/mol for the +2 charged adducts, interacting with the keto-guanine tautomer and the anti-imino adenine tautomer. The surprisingly large interaction energy for the latter structure is due to the strong H-bond between the NH3 ligand group of the metal adduct and the N6 nitrogen atom of the base. PMID- 10819464 TI - The metal binding properties of the zinc site of yeast copper-zinc superoxide dismutase: implications for amyotrophic lateral sclerosis. AB - We have investigated factors that influence the properties of the zinc binding site in yeast copper-zinc superoxide dismutase (CuZnSOD). The properties of yeast CuZnSOD are essentially invariant from pH 5 to pH 9. However, below this pH range there is a change in the nature of the zinc binding site which can be interpreted as either (1) a change in metal binding affinity from strong to weak, (2) the expulsion of the metal bound at this site, or (3) a transition from a normal distorted tetrahedral ligand orientation to a more symmetric arrangement of ligands. This change is strongly reminiscent of a similar pH-induced transition seen for the bovine protein and, based on the data presented herein, is proposed to be a property that is conserved among CuZnSODs. The transition demonstrated for the yeast protein is not only sensitive to the pH of the buffering solution but also to the occupancy and redox status of the adjacent copper binding site. Furthermore, we have investigated the effect of single site mutations on the pH- and redox-sensitivity of Co2+ binding at the zinc site. Each of the mutants H46R, H48Q, H63A, H63E, H80C, G85R, and D83H is capable of binding Co2+ to a zinc site with a distorted tetrahedral geometry similar to that of wild-type. However, they do so only if Cu+ is bound at the copper site or if the pH in raised to near physiological levels, indicating that the change at the zinc binding site seen in the wild-type is conserved in the mutants, albeit with an altered pKa. The mutants H71C and D83A did not bind Co2+ in a wild-type-like fashion under any of the conditions tested. This study reveals that the zinc binding site is exquisitely sensitive to changes in the protein environment. Since three of the mutant yeast proteins investigated here contain mutations analogous to those that cause ALS (amyotrophic lateral sclerosis) in humans, this finding implicates improper metal binding as a mechanism by which CuZnSOD mutants exert their toxic gain of function. PMID- 10819465 TI - Protein control of the formation and decomposition of the CYP119 and CYP101 aryl iron complexes. AB - CYP119, the first thermophilic P450 enzyme, reacts much more slowly than CYP101 (P450cam) with aryldiazenes to give sigma-bonded aryl-iron complexes. The CYP119 complexes are stable anaerobically at 80 degrees C but are readily oxidized by O2 to give the N-arylprotoporphyrin IX regioisomers. The aryl shift can also be initiated in the absence of O2 by K3Fe(CN)6. In contrast, the corresponding CYP101 complexes are insensitive to O2 but decompose at temperatures above 50 degrees C owing to denaturation of the protein. The rate of the CYP119 aryl shift is decreased by electron-withdrawing substituents, with rho = -1.50 for both the O2- and K3Fe(CN)6-dependent reactions. A similar dependence (rho = -0.90) is observed for the K3Fe(CN)6-dependent CYP101 shift. The enthalpies and entropies of activation suggest that the CYP119 and CYP101 K3Fe(CN)6-mediated reactions are similar, but the CYP119 O2-dependent reaction proceeds via a different transition state. In all cases, the rate-determining step is oxidation of the aryl-iron complex. The temperature dependence of the O2- and K3Fe(CN)6-dependent CYP119 shifts provides evidence for temperature-dependent equilibration of two active site conformations. The oxygen sensitivity of the CYP119 aryl-iron complexes, and the temperature dependence of their rearrangement, reflect the unique active site properties of this thermophilic P450 enzyme. PMID- 10819466 TI - The reaction mechanism of nitrosothiols with copper(I). AB - Copper and other transition metal ions and their complexes are catalysts for the decomposition of nitrosothiols. In this way they catalyze the biological functions of nitrosothiols. The kinetics and mechanism of the reaction of two nitrosothiols, S-nitrosothiolactic acid and S-nitrosoglutathione (GSNO), with copper(I) are reported. The kinetics of the reaction of Cu(MeCN)+(n) (n=0-3) with the nitrosothiols were studied. The results indicate that Cu+(aq) is the active species in the GSNO system, with k(Cu+(aq)+GSNO)=(9.4 +/- 2.0) x 10(7) dm3 mol( 1) s(-1). The results also indicate that the Cu(MeCN)+(n) (n=0-3) complexes react with S-nitrosothiolactic acid. Transient species are formed in these processes. The results suggest that these species contain copper(I) and thiol. The results shed light on the catalytic role of copper complexes in the decomposition of S nitrosothiols. PMID- 10819467 TI - Zinc(II) and cadmium(II) metal complexes of thiamine pyrophosphate and 2-(alpha hydroxyethyl)thiamine pyrophosphate: models for activation of pyruvate decarboxylase. AB - Metal complexes of thiamine pyrophosphate (TPP) of the general formula [M2(TPPH)2Cl2]x4H2O (M = Zn2+, Cd2+) were isolated from methanolic solutions and characterized by elemental analysis, FT-IR, and multinuclear NMR spectroscopies. The data provide evidence for the bonding of the metals to the N(1') atom of the pyrimidine ring and to the pyrophosphate group. The stability constant measurements of TPP and 2-(alpha-hydroxyethyl)thiamine pyrophosphate (HETPP) metal complexes in aqueous solution imply the formation of dimeric complex species similar to the isolated solid products. They indicate also that HETPP forms more stable metal complexes than does TPP. To evaluate the coenzyme action of TPP and HETPP metal complexes, enzymic studies have been done using pyruvate decarboxylase apoenzyme. TPP metal complexes do not bind to the apoenzyme, unlike the Zn(II)-HETPP complex which can act as coenzyme. Considering these results, possible functional implications for thiamine involvement in catalysis are discussed. PMID- 10819468 TI - Formation of two types of low-spin heme in horseradish peroxidase isoenzyme A2 at low temperature. AB - Electronic absorption, resonance Raman and EPR spectra are reported for ferric horseradish peroxidase isoenzyme A2 at neutral and alkaline pH together with its imidazole complex at 12 K. The data are compared with those obtained at room temperature. At neutral pH, lowering the temperature induces conformational changes with the formation of two types of low-spin hemes, a bis-histidyl type and a hydroxo type. The transition induced by lowering the temperature is accompanied by a change in the orientation of a vinyl substituent which appears less conjugated to the porphyrin macrocycle than at room temperature. At low temperature the low-spin hemes coexist with a quantum admixed spin species. All the forms are characterized by extremely high resonance Raman frequencies, indicating a contraction of the core size from that of the room temperature species. At alkaline pH, only one low-spin species is observed at both room and low temperatures, with a hydroxo ligand bound to the heme iron. The v(Fe-OH) stretching mode has been assigned at 512 cm(-1), on the basis of the isotopic shift observed in D2O and H2(18)O. This relatively low frequency, together with the anomalous shift observed in deuterium, indicates that the hydrogen bonds between the oxygen atom and the distal residues are stronger than in metmyoglobin, but weaker than those of horseradish peroxidase isoenzyme C. This is in agreement with the lower tetragonality, determined from the EPR g values, of alkaline horseradish peroxidase isoenzyme A2 than of metmyoglobin. PMID- 10819469 TI - A comparative study of galactose oxidase and active site analogs based on QM/MM Car-Parrinello simulations. AB - A parallel study of the radical copper enzyme galactose oxidase (GOase) and a low molecular weight analog of the active site was performed with dynamical density functional and mixed quantum-classical calculations. This combined approach enables a direct comparison of the properties of the biomimetic and the natural systems throughout the course of the catalytic reaction. In both cases, five essential forms of the catalytic cycle have been investigated: the resting state in its semi-reduced (catalytically inactive) and its oxidized (catalytically active) form, A(semi) and A(ox), respectively; a protonated intermediate B; the transition state for the rate-determining hydrogen abstraction step C, and its product D. For A and B the electronic properties of the biomimetic compound are qualitatively very similar to the ones of the natural target. However, in agreement with the experimentally observed difference in catalytic activity, the calculated activation energy for the hydrogen abstraction step is distinctly lower for GOase (16 kcal/mol) than for the mimetic compound (21 kcal/mol). The enzymatic transition state is stabilized by a delocalization of the unpaired spin density over the sulfur-modified equatorial tyrosine Tyr272, an effect that for geometric reasons is essentially absent in the biomimetic compound. Further differences between the mimic and its natural target concern the structure of the product of the abstraction step, which is characterized by a weakly coordinated aldehyde complex for the latter and a tightly bound linear complex for the former. PMID- 10819470 TI - Binding of nitrite and its reductive activation to nitric oxide at biomimetic copper centers. AB - The reactivity of nitrite towards the copper(II) and copper(I) centers of a series of complexes with tridentate nitrogen donor ligands has been investigated. The ligands are bis[(1-methylbenzimidazol-2-yl)methyl]amine (1-bb), bis[2-(1 methylbenzimidazol-2-yl)ethyl]amine (2-bb), and bis[2-(3,5-dimethyl-1 pyrazolyl)ethyl]amine (ddah) and carry two terminal benzimidazole (1-bb, 2-bb) or pyrazole (ddah) rings and a central amine donor residue. While 2-bb and ddah form two adjacent six-membered chelate rings on metal coordination, 1-bb forms two smaller rings of five members. The binding affinity of nitrite and azide to the Cu(II) complexes (ClO4- as counterion) has been determined in solution. The association constants for the two ligands are similar, but nitrite is a slightly stronger ligand than azide when it binds as a bidentate donor. The X-ray crystal structure of the nitrite complex [Cu(ddah)(NO2)]ClO4 (final R=0.056) has been determined: triclinic P1space group, a=8.200(2) A, b=9.582(3) A, c=15.541(4) A. It may be described as a perchlorate salt of a "supramolecular" species resulting from the assembly of two complex cations and one sodium perchlorate unit. The copper stereochemistry in the complex is intermediate between SPY and TBP, and nitrite binds to Cu(II) asymmetrically, with Cu-O distances of 2.037(2) and 2.390(3) A and a nearly planar CuO2N cycle. On standing, solutions of [Cu(ddah)(NO2)]ClO4 in methanol produce the dinuclear complex [Cu(ddah)(OMe)]2(ClO4)2, containing dibridging methoxy groups. In fact the crystal structure analysis (final R=0.083) showed that the crystals are built up by dinuclear cations, arranged on a crystallographic symmetry center, and perchlorate anions. Electrochemical analysis shows that binding of nitrite to the Cu(II) complexes of 2-bb and ddah shifts the reduction potential of the Cu(II)/Cu(I) couple towards negative values by about 0.3 V. The thermodynamic parameters of the Cu(II)/Cu(I) electron transfer have also been analyzed. The mechanism of reductive activation of nitrite to nitric oxide by the Cu(I) complexes of 1-bb, 2-bb, and ddah has been studied. The reaction requires two protons per molecule of nitrite and Cu(I). Kinetic experiments show that the reaction is first order in [Cu(I)] and [H+] and exhibits saturation behavior with respect to nitrite concentration. The kinetic data show that [Cu(2-bb)]+ is more efficient than [Cu(1-bb)]+ and [Cu(ddah)]+ in reducing nitrite. PMID- 10819471 TI - Inhibition of the catecholase activity of biomimetic dinuclear copper complexes by kojic acid. AB - The inhibition of the catechol oxidase activity exhibited by three dinuclear copper(II) complexes, derived from different diaminotetrabenzimidazole ligands, by kojic acid [5-hydroxy-2-(hydroxymethyl)-gamma-pyrone] has been studied. The catalytic mechanism of the catecholase reaction proceeds in two steps and for both of these inhibition by kojic acid is of competitive type. The inhibitor binds strongly to the dicopper(II) complex in the first step and to the dicopper dioxygen adduct in the second step, preventing in both cases the binding of the catechol substrate. Binding studies of kojic acid to the dinuclear copper(II) complexes and a series of mononuclear analogs, carried out spectrophotometrically and by NMR, enable us to propose that the inhibitor acts as a bridging ligand between the metal centers in the dicopper(II) catalysts. PMID- 10819472 TI - Interactions in solution of calcium(II) and copper(II) with nucleoside monophosphates: a calorimetric study. AB - The thermodynamic parameters enthalpy and entropy of the interaction between calcium(II) or copper(II) with 5'-UMP, 5'-CMP, 5'-AMP, 5'-GMP or 5'-IMP in aqueous solution were determined calorimetrically (ionic strength adjusted to 0.1 with tetramethylammonium bromide) at 25 degrees C and pH 7 for Ca(II) or pH 3-5 for Cu(II). The experimental conditions were carefully selected to avoid polynuclear complex formation and nucleotide self-stacking. The calorimetric data confirm the tendency toward macrochelation which was indicated by Sigel after very precise potentiometric studies, and which follows the order Cu(II) > Ca(II) for the metal ions and GMP > IMP > AMP > CMP = UMP for the nucleotides. Macrochelate formation for these metal-nucleoside monophosphate complexes is energetically favorable and entropically unfavorable. PMID- 10819473 TI - Structural evidence for a common zinc binding domain in archaeal and eukaryal transcription factor IIB proteins. AB - X-ray absorption spectroscopy has been used to compare the metal coordination of the N-terminal zinc binding domain of eukaryal human transcription factor (TF) IIB to the previously reported structure of archaeal Pyrococcus furiosus (Pf) TFB. Full length and N-terminal fragments for both PfTFB and human TFIIB were cloned, expressed, and purified. The [C10H] variant of PfTFB was constructed to resemble the metal binding motif of higher eukaryal TFIIB proteins by mutating the second cysteine ligand to a histidine. All five proteins bind zinc in a 1:1 ratio. Zn X-ray absorption spectroscopy of human TFIIB and [C10H]PfTFB mutant are consistent with ZnS3(N,O) ligation, and further suggest that the N/O ligand is an imidazole. PMID- 10819474 TI - Significant changes in the serum levels of IL-6, h-HGF, and type IV collagen 7S during the perioperative period of a hepatectomy: relevance to SIRS. AB - We analyzed the changes in the serum levels of both interleukin-6 (IL-6), human hepatocyte growth factor (h-HGF), and type IV collagen 7S (7S) during the perioperative period of a hepatectomy and evaluated their relationship with systemic inflammatory response syndrome (SIRS). The study subjects consisted of 40 patients who underwent a hepatectomy. In 14 out of 40 patients, postoperative SIRS(+) was observed. Between the SIRS(+) and SIRS(-) cases, there were significant differences in the preoperative values of prothrombin time, hepaplastin test, cholinesterase, and indocyanine green retention at 15 min (P < 0.01). Compared with the SIRS(-) cases, the IL-6, h-HGF, and 7S of the SIRS(+) cases fluctuated in a higher range and remained significantly higher after postoperative day 1 (P < 0.05). Eight out of 14 SIRS(+) patients had postoperative complications. In the 8 SIRS(+) patients with postoperative complications and in the 4 patients in which the SIRS(+) state lasted 3 days or longer, the 7S levels were significantly higher during the perioperative period (P < 0.05). In the SIRS(+) cases, the postoperative levels of IL-6 and h-HGF, as well as pre- and postoperative levels of 7S, were elevated. We therefore consider these levels to be risk factors for complications during the perioperative period of a hepatectomy. PMID- 10819475 TI - The effectiveness of preoperative intraluminal brachytherapy in preventing wall penetration and nodal involvement of rectal carcinomas. AB - A retrospective review was conducted of 115 patients with middle or lower rectal carcinoma, without distant metastasis, who were consecutively treated with preoperative intraluminal brachytherapy and radical surgery (IBT group). Another 115 rectal carcinoma patients treated with surgery alone were reviewed as a control (NIBT group). We investigated whether conventional pathologic stagings were correlated with prognosis. All tumors were diagnosed by the TNM classification. According to the distribution of pathologic staging in the resected specimens, the prevalence of totally ablated tumors (stage T0) in the IBT group was significantly higher than that in the NIBT group, but there were no significant differences in the distribution of other T stages between the two groups. Good local control was achieved for the stage-T3 tumors in the IBT group, similarly to the T < or = 2 tumors in both groups. In the IBT group, the survival period of the patients with stage-T4 tumors even after radiotherapy was significantly shorter than that of those with T3 tumors (P = 0.042), while those with N < or = 2-stage tumors had significantly longer survival periods than those with N3-stage tumors (P = 0.047). These findings demonstrate the usefulness of analyzing clinicopathologic parameters to predict local control and the prolongation of survival. Determining these factors will also help to identify high-risk patients likely to require more effective adjuvant chemotherapy. PMID- 10819476 TI - Pulmonary thromboembolism after surgery for esophageal cancer: its features and prophylaxis. AB - We attempt to clarify the problems of pulmonary thromboembolism (PTE), which occurs less frequently in Japan than in the West, regarding its special perioperative management and prophylaxis for PTE after esophagectomy. We studied 26 patients with PTE following esophagectomy among 1023 patients with esophageal cancer between 1984 and 1997. The presence of embolism was confirmed by pulmonary perfusion scintigraphy. The incidence, diagnosis, and other issues of PTE were all reviewed. The incidence of PTE was 2.5%, with patients showing a biphasic early and late onset. The main symptoms were dyspnea in 19 patients and tachycardia in 17. Scintigraphy demonstrated 154 lesions, 35.7% of which were located in the left lower lobe and 25.3% in the right lower lobe. Treatment mainly consisted of the administration of heparin and urokinase. Four of the 26 patients died. Intermittent pneumatic compression (IPC) with the administration of heparin has been used in our department since 1994 to prevent PTE and this has also helped to decrease the incidence from 3.2% to 0.7%. Because the incidence of PTE following esophagectomy is higher than expected, PTE should be considered whenever hypoxemia of some unknown cause is found. Both early diagnosis and treatment are essential. It is also important to prevent PTE by the use of IPC. PMID- 10819477 TI - The effect of intraoperative colonic lavage with NG-nitro-L-arginine methyl ester (L-NAME) on anastomotic healing in the presence of left-sided colonic obstruction in the rat. AB - The effect of intraoperative colonic lavage with NG-nitro-L-arginine methyl ester (L-NAME) on the healing of colonic anastomosis in the presence of a left-sided obstruction in the rat was investigated. Left-sided colonic obstruction was created in 144 Wistar rats. The obstruction site was excised 24h later and anastomosis was performed after either no irrigation or colonic lavage with either saline, povidone iodine (PI), short-chain fatty acids (SCFA), L-NAME, or glutamine, in 24 animals each. Animals were killed on days 3 and 6, and a 4-cm colonic segment with the anastomosis at the center was excised. Bursting pressure (BP) and hydroxyproline (HP) content were measured. In the saline, PI, and SCFA groups, BP was higher (P < 0.05, P < 0.05, and P < 0.001, respectively) and HP concentration was similar compared with controls. Both the BP and HP concentrations were higher in the glutamine group compared with controls (P < 0.001). BP was lower (P < 0.05) and HP concentration was similar in the L-NAME group compared with the control group. Colonocyte nutrition and tissue perfusion are the mainstays of anastomotic healing. Intraoperative colonic lavage with L NAME suppresses colonic anastomotic healing in the presence of a left-sided obstruction. PMID- 10819478 TI - Enhanced graft healing of high-porosity expanded polytetrafluoroethylene grafts by covalent bonding of fibronectin. AB - The effect of covalent bonding of fibronectin on the patency and graft healing of high-porosity expanded polytetrafluoroethylene (ePTFE) grafts was evaluated. Bilateral carotid grafting was performed in ten mongrel dogs using high-porosity (60 microm) ePTFE grafts, 4 cm in length and 4 mm in internal diameter, that either had been pretreated by the covalent bonding of fibronectin (fibronectin grafts) or were untreated (control grafts). The grafts were harvested 4 to 6 weeks after surgery and subjected to macroscopic and light-microscopic observations. There was no significant difference in patency between the fibronectin grafts and the control grafts with rates of 80% and 70%, respectively. The thrombus-free area score was significantly greater in the fibronectin grafts than in the control grafts, at 86.9% vs 34.0%. Furthermore, the pseudointima was better replaced by fibrous tissue in the fibronectin grafts than in the control grafts, being lined with a layer of endothelial-like cells. More transmural tissue ingrowth was evident in the fibronectin grafts than in the control grafts. The covalent bonding of fibronectin improves graft healing by stimulating transmural tissue ingrowth in high-porosity ePTFE grafts. PMID- 10819479 TI - Gastric carcinoma presenting with extensive extraluminal growth: report of a case. AB - We report a 48-year-old-man with gastric carcinoma presenting with an unusual extraluminal growth. The patient underwent a barium meal examination and gastrofiberscopy because of progressive anemia over 6 months. These examinations revealed a Borrmann type 3 advanced gastric carcinoma of the greater curvature of the antrum. Biopsies showed moderately differentiated tubular adenocarcinoma. The intraoperative findings showed gastric carcinoma associated with extensive extraluminal invasion into the adjacent organs, i.e., the transverse colon and mesocolon. A palliative distal gastrectomy with a partial resection of the transverse colon was performed because of peritoneal dissemination found in the mesocolon and rectovesical pouch. A histological examination of the specimen confirmed adenocarcinoma which had massively infiltrated the transverse colon and mesocolon. His postoperative course was uneventful. However, he died of peritonitis carcinomatosa 9 months later. PMID- 10819480 TI - Gastric perforation caused by a bulimic attack in an anorexia nervosa patient: report of a case. AB - We report a rare case of gastric perforation due to a bulimic attack in a 17-year old girl suffering from anorexia nervosa. She was admitted to our hospital with the chief complaint of abdominal pain following bulimia. Initially, her symptoms were reduced after drainage using a nasogastric tube. Eight hours later, however, she fell into a state of preshock. Abdominal radiography revealed subphrenic free air. We diagnosed the patient as having diffuse peritonitis. At laparotomy, the stomach was dilated and necrotic with perforation. Almost the entire stomach was resected. Postoperatively, the patient recovered uneventfully. We should therefore be aware of this condition when treating patients with anorexia nervosa who complain of abdominal pain. PMID- 10819481 TI - Jejuno-ileal atresia in identical twins: report of a case. AB - Small intestinal atresia occurring in twins is extremely rare. The aims of this study are to report a case of identical twins with different types of jejunal atresia, to review six other pairs of twins, and discuss the etiology. Our case was the seventh in which both twins were diagnosed to have jejuno-ileal atresia. Furthermore, they are also only the sixth and seventh patients known to survive. All the other published cases involved identical twins except one. Three pairs had different types of atresia and four pairs did not have any other anomalies. In addition, to date there have been no further indications of any other members of these families being affected with intestinal atresia. Therefore, we conclude that most such cases might be due to environmental influences during gestation. PMID- 10819482 TI - Appendiceal intussusception induced by tubulovillous adenoma with carcinoma in situ: report of a case. AB - Appendiceal intussusception is an uncommon pathologic condition; however, villous adenoma of the appendix is a distinctly rare entity. We report herein a case of appendiceal intussusception induced by tubulovillous adenoma with carcinoma in situ. A 67-year-old man was admitted to our hospital with a 1-year history of lower abdominal pain for investigation. Barium enema showed a filling defect with an irregular surface in the cecum, and colonoscopy revealed a cecal tumor with a granular surface. Pathological examination of biopsy samples revealed tubulovillous adenoma with well-differentiated adenocarcinoma, and a diagnosis of cecal cancer in tubulovillous adenoma was made. Surgery was performed and the resected specimen was found to contain a tumor arising from the appendix. The tumor was 5.5 x 4.5 cm in size in the cecal cavity, and the appendix had invaginated into the cecum at its base. The cut surface of the appendix showed the villous tumor filling the appendiceal lumen and projecting into the cecal cavity. Microscopic examination revealed well-differentiated adenocarcinoma in tubulovillous adenoma. To the best of our knowledge, this is the first report of appendiceal intussusception caused by tubulovillous adenoma with carcinoma of the appendix. PMID- 10819483 TI - Ileal duplication presenting as perforation: report of a case. AB - We describe herein the case of a 10-year-old boy in whom generalized peritonitis was caused by perforation of a tubular communicating ileal duplication cyst. Alimentary tract duplication cysts are rare congenital malformations, found primarily in children under the age of 15 years. The perforation was caused by heterotopic gastric mucosa within the duplication, giving rise to peptic ulceration in the adjacent intestinal mucosa. The presence of heterotopic gastric tissue is a primary cause of perforation that has been reported in as many as one third of patients with small intestine duplications; however, the detection of such ectopic tissue is time-consuming, and there is no readily available method of diagnosing tubular duplications. Although very few patients present with peritonitis as the initial manifestation, the possibility should be borne in mind when diagnosing and planning therapy for such emergencies, particularly in children. PMID- 10819484 TI - Hemoperitoneum secondary to exophytic leiomyoma: report of a case. AB - We report herein the case of a 63-year-old male with hemoperitoneum secondary to exogastric leiomyoma. The patient had been receiving anticoagulation therapy for a cerebral embolism and complained of sudden, severe abdominal pain. A sonogram and computed tomography scan showed an exogastric mass and massive ascites. A peritoneal puncture proved the presence of an intraperitoneal hemorrhage. An emergency laparotomy revealed a pedunculated bleeding tumor, thus confirming the preoperative diagnosis of a ruptured exogastric tumor. A microscopic analysis of the excised tumor demonstrated gastric leiomyoma. Other authors have reported hemoperitoneum secondary to gastric myogenic tumors, but no cases of leiomyomas could be found in the literature. PMID- 10819485 TI - Idiopathic segmental infarction of the greater omentum successfully treated by laparoscopy: report of case. AB - Idiopathic or spontaneous segmental infarction of the greater omentum (ISIGO) is a rare cause of acute right-sided abdominal pain. The symptoms simulate acute appendicitis in 66% of cases and cholecystitis in 22%. Progressive peritonitis usually dictates laparotomy, and an accurate diagnosis is rarely made before surgery. The etiology of the hemorrhagic necrosis is unknown, but predisposing factors such as anatomic variations in the blood supply to the right free omental end, obesity, trauma, overeating, coughing, and a sudden change in position may play a role in the pathogenesis. We present herein the case of a 37-year-old man in whom ISIGO, precipitated by obesity and overeating, was successfully diagnosed and treated by laparoscopy. Resection of the necrotic part of the greater omentum is the therapy of choice, and ensures fast recovery and pain control. Serohemorrhagic ascites is a common finding in ISIGO, and careful exploration of the whole abdominal cavity should be performed. The laparoscopic approach allows both exploration and surgical intervention. PMID- 10819486 TI - Generalized peritonitis due to spontaneously perforated pyometra presenting as pneumoperitoneum: report of a case. AB - We report a rare case of generalized peritonitis due to a ruptured pyometra in an 86-year-old woman, and also conduct a review of the previous Japanese literature. The patient presented with muscle guarding and rebound tenderness. Computed tomography (CT) disclosed a cystic mass in the peritoneal cavity, in which an air fluid level was noted. Pneumoperitoneum around the uterus due to gas production of anaerobic bacteria was noted on a CT. At laparotomy, the uterus was markedly enlarged with a necrotic area on the uterine fundus, which was found to be perforated. A supravaginal hysterectomy and drainage were performed. We found only eight cases of a ruptured pyometra presenting as pneumoperitoneum in the Japanese literature between 1977 and 1999. The most common cause of pneumoperitoneum is a perforation of the gastrointestinal tract. However, other possible causes, as seen in our patient, should also be taken into consideration. Although it is rare, a perforated pyometra should therefore also be considered when elderly women present with acute abdominal pain. PMID- 10819487 TI - Tuberculous peritonitis defying diagnosis: report of a case. AB - A case of tuberculous peritonitis, which has been scarcely encountered in clinical practice in recent years, is reported. A 32-year-old man was admitted to our hospital complaining of abdominal fullness, anorexia, and a 15 kg weight loss. His abdomen was distended. There was neither any previous history nor recent contact with tuberculosis. The laboratory data indicated increased C reactive protein and erythrocyte sedimentation rate, but the white blood cell count was normal. A chest X-ray examination revealed no abnormalities. Abdominal X-ray showed scattered, small-intestinal gas shadows. Abdominal computed tomography scanning revealed a diffuse thickening of the dilated bowel wall, mainly adjacent to the mesentery. After a detailed examination a diagnosis of peritonitis carcinomatosa of unknown origin was suspected, and an exploratory laparotomy was done. Severe adhesions between the parietal peritoneum and the bowel were found. An excisional biopsy specimen was taken from the peritoneum, and a diagnosis of tuberculosis was thus made. Triple therapy with isoniazid, rifampicin, and kanamycin was started, and both the intestinal obstruction and anorexia were thus resolved. PMID- 10819488 TI - Rib metastasis appearing 8 years after surgery for lung cancer: report of a case. AB - A 39-year-old man underwent a right upper lobectomy and lymph node dissection for right lung adenocarcinoma on March 7, 1988. He was referred for an evaluation of a systemic bone scintillation scan on October 29, 1996. A hot spot at the right fourth rib was recognized. After performing needle aspiration cytology, a diagnosis of adenocarcinoma was made. This case was considered to be rib metastasis occurring 8 years after surgery for lung cancer. In general, regular follow-up examinations are performed for at least 5 years after surgery; however, surgeons should also keep such late metastatic cases in mind. PMID- 10819489 TI - Successful treatment of a bronchial inflammatory pseudotumor by bronchoplasty in an 8-year-old boy: report of a case. AB - We report herein the rare case of an 8-year-old boy in whom an inflammatory pseudotumor of the upper lobe bronchus of the right lung was successfully treated by bronchoplasty. A bronchoscopy was initially performed to investigate the cause of pulmonary atelectasis in the right upper lobe, which revealed a tumor in the right main bronchus. Thus, a thoracotomy followed by bronchotomy of the right main bronchus was carried out. The tumor was seen to have polypoid protrusion into the right main bronchus at the orifice of the upper lobe. A bronchoplasty with a wedge resection of the right main bronchus and right upper lobectomy was carried out, effectively preserving right pulmonary function. Histological examination confirmed the diagnosis of an inflammatory pseudotumor. The patient had an uneventful postoperative course and has been free of recurrence for 3 years since his operation. PMID- 10819490 TI - Bilateral recurrent pneumothorax complicating chemotherapy for pulmonary metastatic breast ductal carcinoma: report of a case. AB - Secondary spontaneous pneumothorax (SSP) is a rare complication of chemotherapy for pulmonary metastases and to the best of our knowledge, only 28 cases have been described, most of which occurred in patients with osteosarcoma or germ cell tumors. We present herein the case of a 56-year-old woman in whom bilateral and recurrent SSP was caused by the rupture of pulmonary lacunae induced by chemotherapy, given for bilateral lung metastases secondary to breast carcinoma. Our experience of this case led us to conclude that: patients with pulmonary metastases may develop bilateral and/or recurrent pneumothoraces following chemotherapy; computed tomography scan is essential for defining the cause of SSP; and closed chest tube drainage remains the therapy of choice, while chemical pleurodesis may also be used to prevent recidivant SSP. PMID- 10819491 TI - Castleman's disease in the posterior mediastinum: report of a case. AB - We report a case of Castleman's disease which developed in the posterior mediastinum, with a review of the Japanese literature. A 19-year-old female patient with asthma was pointed out to have an abnormal shadow in the right posterior mediastinum on chest X-ray. We had tried to perform thoracoscopic surgery for this tumor, but we had to convert the surgical approach from thoracoscopy to a thoracotomy because of both tight adhesion of the tumor and muscle and profuse bleeding from the tumor. The tumor was diagnosed to be the hyaline vascular type of Castleman's disease histopathologically. In the Japanese literature, Castleman's disease, which develops in the posterior mediastinum, has been reported to often accompany tight adhesion between the tumor and surrounding tissue, and profuse bleeding thus cannot be avoided at surgery. The large amount of bleeding observed during the surgery of a patient with Castleman's disease in the posterior mediastinum may be due to tight adhesion and hypervascularity of the tumor. Therefore, care should be exercised in choosing the surgical approach if Castleman's disease is suspected in cases of posterior mediastinal tumor. PMID- 10819492 TI - Blunt carotid artery injury after accidental neck compression: report of a case. AB - Almost all cases of carotid artery injury are precipitated by a high-energy impact such as motor vehicle accidents or gunshot wounds, and are usually diagnosed using angiography. We report herein a case of carotid artery injury induced by a low-energy insult with rare clinical signs which was diagnosed using ultrasonography as well as angiography. A 37-year-old man sustained an accidental compression of the neck and was transferred to our emergency room. Horner's syndrome and phrenic nerve palsy were detected on the left side. Ultrasonography demonstrated two sites of injury with an intimal flap of the distal left common carotid artery as well as angiography. The patient was placed on anticoagulants and was discharged on the 10th hospital day with both Horner's syndrome and phrenic nerve palsy. This case suggests that surgeons should investigate any possible carotid artery injury, even after low-velocity injuries such as compression of the neck, and therefore an ultrasonic examination should be performed at the initial evaluation and at follow-up studies. In addition, further investigations are also called for to investigate the utility of anticoagulation in the treatment of carotid artery injury. PMID- 10819493 TI - Surgical repair of a solitary deep femoral arterial aneurysm: report of two cases. AB - We report herein two cases of a solitary deep femoral arterial aneurysm that were successfully treated by aneurysmectomy and ligation of the deep femoral artery. The patients were a 69-year-old man and a 73-year-old man, both of whom presented with localized pain and a pulsatile mass in the femoral area showing rapid enlargement. Ultrasonography and arteriography confirmed the diagnosis and revealed no evidence of aneurysm or occlusive disease in the other arteries. Aneurysmectomy and ligation of the deep femoral artery were performed, and no ischemic symptoms or thrombus developed postoperatively in either patient. Preoperative arteriography is essential to delineate not only the state of the aneurysm, but also that of the distal arterial tree. An emergency operation is also mandatory due to the rapid enlargement of this type of aneurysm, even if rupture occurs. PMID- 10819494 TI - Autografting with peripheral blood CD34-positive cells following high-dose chemotherapy against breast cancer. AB - We report autologous CD34+ cell transplantation performed in 3 cases of recurrent breast cancer. The hematological recovery in these cases was assessed by comparing with that in the previous cases of autologous hematopoietic stem cell transplantation performed with the same high-dose chemotherapy regimen. Patient 1 was a 32-year-old woman with pulmonary and skeletal metastases; patient 2, a 55 year-old woman with pulmonary metastases; and patient 3, a 48-year-old woman with hepatic metastases. On day 1, cyclophosphamide 1000 mg/m2 and epirubicin 130 mg/m2 were administered concurrently with granulocyte colony-stimulating factor, and peripheral blood stem cells were harvested on days 14-16. These stem cells were processed using anti-CD34 monoclonal antibody and an immunomagnetic bead device, Isolex 300i. The high-dose chemotherapy regimen consisted of cyclophosphamide 2000 mg/m2/day, div, and thiotepa 200 mg/m2/day, div on day -5, 4, and -3. The harvested CD34+ cells numbered 3.9 +/- 2.8 x 10(6)/kg (range: 0.73 7.8/10(6)/kg), and the CFU-GM, 8.3 +/- 5.6 x 10(5)/kg (range: 1.2-15.1/10(5)/kg). After the separation, the percent of CD34+ cells was 81.9 +/- 11.6% (range: 65.8 96.4%), the CD34+ cell yield, 71.8 +/- 30.2% (range: 46.0-129.6%), and the CFU-GM yield, 48.9 +/- 9.1% (range: 35.3-62.0%). At the time of transplantation, the number of nucleated cells was 0.55 +/- 0.31 x 10(5)/kg, and that of CFU-GM, 31.2 +/- 17.8 x 10(5)/kg. Comparison of the hematological recovery in these three cases with that in patients receiving an identical high-dose chemotherapy regimen revealed recovery rates significantly faster than in patients having bone marrow transplants, and approximately identical with that in peripheral blood stem cell transplantation cases. PMID- 10819496 TI - Delayed ischemic neurological deficit of short duration in patients with subarachnoid hemorrhage. AB - We retrospectively examined the duration of delayed ischemic neurological deficits (DIND) attributed to vasospasm following subarachnoid hemorrhage (SAH) in order to delineate the nature and involved factors. Among 605 patients with SAH, 201 developed DIND, and 137 of these had undergone early aneurysmal obliteration. In these 137, duration of DIND was clearly determined in 67 cases. Hypervolemic therapy was instituted only after the onset of DIND. In the 67 patients, the mean duration of DIND was 5.2 days (ranging from 2 to 13 days); 22 patients had DIND lasting only 2 to 3 days, 26 patients had DIND lasting 4 to 6 days, and in 19, DIND lasted 7 to 13 days. Clinical factors associated with short DIND duration (2 to 3 days) as determined by multivariate analysis included internal carotid artery or middle cerebral artery aneurysm, age under 60, and a good World Federation of Neurological Surgeons grade on admission. No patients developed pulmonary edema from hypervolemic therapy, and the outcomes of all 67 patients were extremely favorable. In SAH patients developing DIND after early aneurysmal surgery, 16% (22 of 137) had a documented brief duration of DIND. We believe DIND rapidly improved and resolved with hypervolemic therapy because antecedent brain damage from SAH or surgical manipulation had been minimal. PMID- 10819495 TI - Theoretical background for inward rectification. AB - Theoretical background has been reviewed for inward rectification due to a potassium current termed IRK. The Eyring rate theory in which the thermodynamic rate coefficient for chemical reactants (channels and ions in this case) can be described in terms of energy barriers for potassium ions can mimic not only the polarity and degree of rectification but also the voltage-dependence of the barium-induced IRK block. The model predicts that the blocking site locates 30 70% depth from the outer margin of the IRK channel. PMID- 10819497 TI - A case of gastrointestinal stromal tumor of the stomach. AB - In a 55-year-old man, a tumor about 3 cm in diameter was detected in the upper abdomen by abdominal ultrasound screening during follow-up of chronic hepatitis C discovered in 1990. There were no symptoms and no abnormalities on physical examination. Tests for tumor markers were negative. By barium meal and gastroscopy, submucosal tumor was found on the lesser curvature of the stomach, with bridging fold in the absence of central ulceration. Biopsy revealed no tumor tissue. Under the diagnosis of submucosal tumor of the stomach, either a leiomyoma or leiomyosarcoma, partial resection of stomach was performed. Direct invasion of the surrounding organs, lymph node metastasis or distant metastasis was not observed grossly in the operation. Histologic examination of the resected specimen revealed proliferation of spindle cells and oval cells in an interlacing pattern. Immunohistochemistry for CD34, vimentin and c-kit protein was strongly positive, while smooth muscle actin, S-100 protein, desmin and p53 protein were negative. The proliferating cell nuclear antigen index was about 50%, while the MIB-1 index was < or = 1%. From these findings, this tumor was diagnosed as a gastrointestinal stromal tumor of the uncommitted type. PMID- 10819498 TI - Medical students and infection control: risks and precautions. AB - PURPOSE: To investigate factors associated with medical students' use of infection control measures to protect themselves from HIV and other blood-borne infections. METHODS: All clinical students in the academic year of 1993-94 at the University of Washington, Seattle, were surveyed on the percent of time they use Universal Precautions (UPs), their sense of control and concern over HIV risk, estimates of occupational HIV risk and career plans. RESULTS: Responses of 294 students (86%) were studied. Most students use UPs most of the time but only 25% do so universally. Women (88.1%) use UPs more often than men (82.8%). Use of UPs was not related to the factors that actually predict risk of infection: incidence of exposure accidents and prevalence of HIV among patients. UP use was not associated with demographic characteristics of the student, specialty choice or level of concern about HIV. Students with greater sense of control over their risk reported greater use of UP. Those who plan to restrict patient services because of HIV risk use UP less often than others. CONCLUSION: Students use UPs but could do more to protect themselves. Infection control among young clinicians may be improved by education about the reality of the risk and the efficacy of precautionary measures. PMID- 10819499 TI - Estrogens and progestins in mammary development and neoplasia. Introduction. PMID- 10819500 TI - Are the TDLU of the human the same as the LA of mice? PMID- 10819501 TI - Mammary gland growth and development from the postnatal period to postmenopause: ovarian steroid receptor ontogeny and regulation in the mouse. AB - Ovarian steroid hormones play a critical role in regulating mammary gland growth and development. The mammary gland sequentially acquires and cyclically exhibits proliferative responses to estrogen and/or progesterone from birth to postmenopause. The focus of this review is to present our current understanding of estrogen and progesterone receptor distribution in epithelial and stromal cells and their functions in relation to mammary gland development. Insights gained from the study of the normal mammary gland are relevant to our understanding of the conditions which may predispose women to the development of breast cancer as well as to alterations in hormonal regulation that occur in breast cancer. PMID- 10819502 TI - Estrogen responsiveness and control of normal human breast proliferation. AB - Our understanding of the hormonal control of the proliferation of normal human breast epithelium is still surprisingly meager. However, the results of a number of recent studies have confirmed that estrogen is the major steroid mitogen for the luminal epithelial cell population (the usual targets for neoplastic transformation). Estrogen seemingly exerts its effects on cell division indirectly as there is complete dissociation between the population of luminal epithelial cells expressing the estrogen receptor (ER)4 and those that proliferate. We suggest that the ER-negative proliferating cells represent a precursor or stem cell population that differentiates to ER-containing, nonproliferative cells. In turn, these ER-positive cells act as 'estrogen sensors' and transmit positive or negative paracrine growth signals to the precursor cells depending on the prevailing hormonal environment. As yet there is no direct evidence supporting this hypothesis but we suggest ways in which it may be obtained. The implication of these studies is that inhibition of luminal epithelial proliferation with tamoxifen or pure antiestrogens or by preventing ovarian steroid secretion should be an effective strategy for the prevention of breast cancer. In addition, we may be able to predict the risk of breast cancer in an individual by measuring the intrinsic estrogen sensitivity of her breast epithelium. Finally, study of the paracrine mechanisms of growth control in the normal human breast may provide new, more specific, therapeutic targets for breast cancer prevention. PMID- 10819503 TI - Use of the autopsy to study ontogeny and expression of the estrogen receptor gene in human breast. AB - Historically, autopsies have been a valuable resource for morphologic studies of the human breast, and have been used in conjunction with radiologic evaluation and epidemiology to provide information about population prevalence of pre malignant and malignant disease. More recently, well-preserved post-mortem breast tissue has also been used to evaluate the status of genes and their expression. Using molecular techniques and immunohistochemistry, quantitation of gene expression and localization of proteins of hormone receptors, growth factor receptors, growth factors, cell-proliferation related antigens and proto oncogenes can be evaluated in autopsy-derived breast tissue. Expression of the estrogen receptor (ER)3 gene at the mRNA and protein product levels has been evaluated in breast tissue from infants, children, adolescent girls, and adult women in various phases of the menstrual cycle, pregnancy and post-menopausal. The results of these studies support a role for the ER gene in early as well as pubescent breast development, and also in normal cyclical and abnormal cell proliferation in the terminal-duct-lobular-units (TDLU) of adult women. PMID- 10819504 TI - Role of hormones in mammary cancer initiation and progression. AB - Breast cancer, the most frequent spontaneous malignancy diagnosed in women in the Western world, is a classical model of hormone dependent malignancy. There is substantial evidence that breast cancer risk is associated with prolonged exposure to female hormones, since early onset of menarche, late menopause, hormone replacement therapy and postmenopausal obesity are associated with greater cancer incidence. Among these hormonal influences a leading role is attributed to estrogens, either of ovarian or extra-ovarian origin, as supported by the observations that breast cancer does not develop in the absence of ovaries, ovariectomy causes regression of established malignancies, and in experimental animal models estrogens can induce mammary cancer. Estrogens induce in rodents a low incidence of mammary tumors after a long latency period, and only in the presence of an intact pituitary axis, with induction of pituitary hyperplasia or adenomas and hyperprolactinemia. Chemicals, radiation, viruses and genomic alterations have all been demonstrated to have a greater tumorigenic potential in rodents. Chemical carcinogens are used to generate the most widely studied rat models; in these models hormones act as promoters or inhibitors of the neoplastic process. The incidence and type of tumors elicited, however, are strongly influenced by host factors. The tumorigenic response is maximal when the carcinogen is administered to young and virgin intact animals in which the mammary gland is undifferentiated and highly proliferating. The atrophic mammary gland of hormonally-deprived ovariectomized or hypophysectomized animals does not respond to the carcinogenic stimulus. Administration of carcinogen to pregnant, parous or hormonally treated virgin rats, on the other hand, fails to elicit a tumorigenic response, a phenomenon attributed to the higher degree of differentiation of the mammary gland induced by the hormonal stimulation of pregnancy. In women a majority of breast cancers that are initially hormone dependent are manifested during the postmenopausal period. Estradiol plays a crucial role in their development and evolution. However, it is still unclear whether estrogens are carcinogenic to the human breast. The apparent carcinogenicity of estrogens is attributed to receptor-mediated stimulation of cellular proliferation. Increased proliferation could result in turn in accumulation of genetic damage and stimulation of the synthesis of growth factors that act on the mammary epithelial cells via an autocrine or paracrine loop. Alternatively estrogens may induce cell proliferation through negative feedback by removing the effect of one or several inhibitory factors present in the serum. Multidisciplinary studies are required for the elucidation of the mechanisms responsible for the initiation of breast cancer. Understanding of such mechanisms is indispensable for developing a rational basis for its prevention and control. PMID- 10819505 TI - Estrogen and progestin regulation of cell cycle progression. AB - Estrogens and progesterone, acting via their specific nuclear receptors, are essential for normal mammary gland development and differentiated function. The molecular mechanisms through which these effects are mediated are not well defined, although significant recent progress has been made in linking steroid hormone action to cell cycle progression. This review summarizes data identifying c-myc and cyclin D1 as major downstream targets of both estrogen- and progestin stimulated cell cycle progression in human breast cancer cells. Additionally, estrogen induces the formation of high specific activity forms of the cyclin E Cdk2 enzyme complex lacking the cyclin-dependent kinase (CDK)3 inhibitor, p21. The delayed growth inhibitory effects of progestins, which are likely to be prerequisites for manifestation of their function in differentiation, also involve decreases in cyclin D1 and E gene expression and recruitment of CDK inhibitors into cyclin D1-Cdk4 and cyclin E-Cdk2 complexes. Thus estrogens and progestins affect CDK function not only by effects on cyclin abundance but also by regulating the recruitment of CDK inhibitors and, as yet undefined, additional components which determine the activity of the CDK complexes. These effects of estrogens and progestins are likely to be major contributors to their regulation of mammary epithelial cell proliferation and differentiation. PMID- 10819506 TI - Estrogen receptor variants. AB - Estrogen receptor (ER)3 gene expression in breast epithelium is an intricately regulated event. The human ER gene is transcribed from at least three different promoters which are expressed in a cell- and tissue-specific manner, and result in mRNA isoforms with unique 5'-untranslated exons. The ER is overexpressed in about two thirds of breast tumors, and even in early premalignant breast lesions compared with adjacent normal breast epithelium. Furthermore, normal breast epithelium as well as breast cancer tissue contains alternatively spliced ER mRNA variants where single or multiple exons are skipped. It is still unclear if any or all of the ER mRNA splicing variants are translated in vivo, and if a change in the balance of ER variants could effect tumor development and progression to hormone-independent growth. Although infrequent in primary breast cancer, single amino acid changes within the ER in metastatic disease which might influence cell proliferation may also contribute to neoplastic progression of the mammary epithelium. PMID- 10819507 TI - The loss of estrogen and progesterone receptor gene expression in human breast cancer. AB - Hormone responsiveness is a critical determinant of breast cancer progression and management, and the response to endocrine therapy is highly correlated with the estrogen receptor (ER)3 and progesterone receptor (PR) status of tumor cells. Thus, key areas of study in breast cancer are those mechanisms that regulate ER and PR expression in normal and malignant breast tissues. One-third of all breast cancers lack ER and PR; these conditions are associated with less differentiated tumors and poorer clinical outcome. In addition, approximately one-half of ER positive tumors lack PR protein and patients with this phenotype are less likely to respond to hormonal therapies than those whose tumors express both receptors. Since PR is induced by ER; its presence is a marker of a functional ER. In this review, we will discuss possible mechanisms for loss of ER and PR gene expression, especially structural changes within each gene including deletions, polymorphisms or methylation. Improved understanding of the pathways that lead to loss of ER and/or PR proteins should allow the development of better predictive indicators as well as novel therapeutic approaches to target these hormone independent cancers. PMID- 10819508 TI - Role of the androgen receptor in human breast cancer. AB - Although the androgen receptor (AR)3 is often co-expressed with the estrogen receptor (ER) and progesterone receptor (PR) in human breast tumors, its role in breast cancer is poorly understood. Specific growth stimulatory and inhibitory actions of androgens have been described in human breast cancer cell lines. The mechanisms by which androgens exert these contrasting growth effects are unknown. A commonly utilized second line therapy for the treatment of advanced breast cancer is high dose medroxyprogesterone acetate (MPA). Although MPA, a synthetic progestin, was thought to act exclusively through the PR, the androgenic side effects observed in women taking MPA suggest that its action may also be mediated in part by the AR. In support of this hypothesis, the level of AR measured by radioligand binding in primary breast tumors was correlated with the duration of response to MPA treatment following failure of tamoxifen therapy. Recent data suggest that the presence of structurally altered AR in breast cancers may account for unresponsiveness to MPA in some of these cases. Further studies are warranted to determine the role of AR mediated pathways in regulating breast tumor growth. In particular, identification of androgen-regulated genes may lead to new possibilities for the hormonal treatment of breast cancer. PMID- 10819509 TI - Helene Smith: a memorial. PMID- 10819510 TI - The cell biology of milk secretion: historical notes. Introduction. PMID- 10819511 TI - Tight junction regulation in the mammary gland. AB - Tight junctions form a narrow, continuous seal that surrounds each endothelial and epithelial cell at the apical border, and act to regulate the movement of material through the paracellular pathway. In the mammary gland, the tight junctions of the alveolar epithelial cells are impermeable during lactation, and thus allow milk to be stored between nursing periods without leakage of milk components from the lumen. Nonetheless mammary epithelial tight junctions are dynamic and can be regulated by a number of stimuli. Tight junctions of the mammary gland from the pregnant animal are leaky, undergoing closure around parturition to become the impermeable tight junctions of the lactating animal. Milk stasis, high doses of oxytocin, and mastitis have been shown to increase tight junction permeability. In general changes in tight junction permeability in the mammary gland appear to be the results of a state change and not assembly and disassembly of tight junctions. Both local factors, such as intramammary pressure and TGF-beta, and systemic factors, such as prolactin, progesterone, and glucocorticoids, appear to play a role in the regulation of mammary tight junctions. Finally, the tight junction state appears to be closely linked to milk secretion. An increase in tight junction permeability is accompanied by decrease in the milk secretion rate, and conversely, a decrease in tight junction permeability is accompanied by an increase in the milk secretion rate. PMID- 10819513 TI - Origin and secretion of milk lipids. AB - The cream fraction of milk comprises droplets of triacylglycerol coated with cellular membranes. In this review, we discuss how these droplets are formed and secreted from mammary epithelial cells during lactation. This secretory system is especially interesting because the assembled lipid droplets are secreted from the cytoplasm enveloped by cellular membranes. In other cells, such as hepatocytes and enterocytes, lipid is secreted by exocytosis from membrane-bounded compartments of the secretory pathway. Milk lipids originate as small droplets of triacylglycerol, synthesized in or on the surfaces of rough endoplasmic reticulum (ER)4 membranes. These droplets are released into the cytoplasm as microlipid droplets (MLDs) with a surface coat of protein and polar lipid. MLDs may fuse with each other to form larger cytoplasmic lipid droplets (CLDs). Droplets of varying size, are transported to the apical cytoplasm by unknown mechanisms and are secreted from the cell coated with an outer bilayer membrane. CLDs may increase in size in all regions of the cell, especially at the plasma membrane during secretion. Two possible mechanisms for lipid secretion have been proposed: an apical mechanism, in which lipid droplets are enveloped with apical plasma membrane, and a secretory-vesicle mechanism, in which fat droplets are surrounded by secretory vesicles in the cytoplasm and are released from the surface by exocytosis from intracytoplasmic vacuoles. A combination of both mechanisms may be possible. Following secretion, a fraction of the membrane surrounding the globules may be shed from the droplets and give rise to membrane fragments in the skim milk phase. PMID- 10819512 TI - Mammary gland membrane transport systems. AB - The secretion of milk depends on the activity of a large number of membrane transport systems located on the apical and basolateral membranes of mammary secretory cells. It follows that a thorough knowledge of individual mammary tissue membrane transport systems is required if we are to fully understand the process of milk secretion. The distribution of the transporters between the apical and basolateral poles of the mammary epithelium must be asymmetrical given that the mammary gland is capable of vectorial transport. This is particularly evident in the case of glucose and amino acid transport systems: the transport mechanisms for these compounds are predominantly situated in the blood-facing aspect of the secretory cells. In addition. it is apparent that there is a polarized distribution of transport systems (carriers and channels) which accept sodium, potassium, chloride, phosphate, and calcium as substrates. PMID- 10819514 TI - Secretion of milk proteins. AB - Mammary epithelial cells secrete milk proteins in a polarized manner from their apical surface during lactation. These secreted proteins are either synthesized by the mammary cells or are transported by transcytosis from blood plasma. The intracellular trafficking pathways by which milk proteins are secreted are known in general outline. In this review the basic cell biology of the mammary epithelial cell secretory pathway is considered in relation to what is known in more detail for other cell types. In addition, potential points of control of protein secretion are examined. The secretory biology of mammary epithelial cells has not been characterized extensively in recent years and, while some aspects are well understood, other key issues, which still remain to be resolved, have been highlighted. PMID- 10819515 TI - Epithelial transcytosis of immunoglobulins. AB - Transcytosis plays a central role in the immunological functions of epithelia, including the sampling of antigens that enter the body via the digestive, respiratory and urogenital tracts and their presentation to underlying lymphoid tissues, the secretion of specific immunoglobulins required for the immune protection of mucosal surfaces and the transfer of maternal immunoglobulins to the fetus or newborn, providing the latter with passive immunity for the first weeks of independent life. PMID- 10819516 TI - Transferrin and prolactin transcytosis in the lactating mammary epithelial cell. AB - The mammary epithelial cell ferries constituents originating from blood and from stromal cells, into milk, by transcytosis. Morphological analysis of a membrane marker of transcytosis in the lactating mammary epithelial cell showed that very rapid endocytosis of surface membrane occurs from both the basolateral and the apical side of the cell. In both cases, membrane trafficking between endosomes and the Golgi complex allows communication between the endocytic and the biosynthetic pathways. Transferrin and prolactin are internalized in mammary cells and transported through multivesicular bodies and Golgi stacks. They are released into milk via different types of secretory vesicles, prolactin being carried in secretory vesicles containing casein micelles. Consequences of the intracellular transport of these proteins and physiological benefits for cell function are discussed. PMID- 10819517 TI - Beta1,4-galactosyltransferase and lactose biosynthesis: recruitment of a housekeeping gene from the nonmammalian vertebrate gene pool for a mammary gland specific function. AB - Beta1,4-galactosyltransferase (beta4GalT-I) is a constitutively expressed trans Golgi enzyme, widely distributed in vertebrates, which synthesizes the beta4-N acetyllactosamine structure commonly found in glycoconjugates. In mammals beta4GalT-I has been recruited for a second biosynthetic function, the production of lactose; this function takes place exclusively in the lactating mammary gland. In preparation for lactose biosynthesis, beta4GalT-I enzyme levels are increased significantly. We show that mammals have evolved a two-step mechanism to achieve this increase. In step one there is a switch to the use of a second transcriptional start site, regulated by a stronger, mammary gland-restricted promoter. The transcript produced is distinguished from its housekeeping counterpart by the absence of approximately 180 nt of 5'-untranslated sequence. In step two, this truncated transcript is translated more efficiently, relative to the major transcript expressed in all other somatic tissues. PMID- 10819518 TI - Developmental and hormonal regulation of protein N-glycosylation in the mammary gland. AB - Glycosylation represents the most common conjugation of both membrane-bound and secreted proteins of animal cells. Among the different types of glycosylation, the N-linked attachment of sugars to the polypeptide backbone is by far the most abundant modification. The biosynthesis of the precursor carbohydrate unit of these proteins is initiated by a stepwise assembly of Glc3Man9GlcNAc2P-P-Dol in the dolichol cycle, its transfer en bloc to the nascent polypeptide in the rough endoplasmic reticulum (RER), followed by excision of the glucosyl residues by processing-specific enzymes, glucosidase I and II, also resident in the endoplasmic reticulum. Additional posttranslational modifications of the carbohydrate moiety in the RER, Golgi, and trans-Golgi network, differ for individual glycoproteins for the completion of final products as high mannose, complex or hybrid glycoproteins en route to their final destinations in the secretory pathway. The enzyme GlcNAc-1-P transferase (GPT) catalyzes the first and committed step, i.e., the transfer of GlcNAc-1-P from UDP-GlcNAc to Dol-P to form GlcNAc-P-P-Dol, in the assembly of the oligosaccharide precursor. Glucosidase I triggers the maturation phase by clipping the distal alpha 1,2 linked Glc residue on the incipient glycoprotein. The critical juxtaposition of the two enzymes in the multistep pathway makes them excellent candidates for the overall regulation of protein N-glycosylation. The highly elevated needs of glycosylation during lactation demand regulation of glycosylation in the gland over and above the levels in the quiescent, virgin and postlactating, regressed gland. PMID- 10819519 TI - The mammary gland as a bioreactor: expression, processing, and production of recombinant proteins. AB - A variety of transgenic animal species are being used to produce recombinant proteins. The general approach is to target the expression of the desired protein to the mammary gland using regulatory elements derived from a milk protein gene and then collect and purify the product from milk. Promoter sequences from a number of different milk protein genes have been used to target expression to the mammary gland, although significant problems remain with regard to achieving transgene expression levels consistent with commercial exploitation. The mammary gland appears to be capable of carrying out the complex posttranslational modifications. such as glycosylation and gamma-carboxylation required for the biological activity and stability of specific proteins. Effective purification protocols have been established and products produced by this route have now entered clinical trials. PMID- 10819520 TI - Modification and repression of genes expressed in the mammary gland using gene targeting and other technologies. AB - Transgenic experiments using oocyte micro-injection methodology are often performed in order to target expression of a foreign gene in a specific tissue or, to a lesser extent, to study the regulation of gene expression. However, the isolation of embryonic stem cells in mice and the development of antisense and ribozyme technologies have allowed more subtle alterations of endogenous gene expression to be achieved. The mammary gland is one of the few organs able to undergo several cycles of development, differentiation and apoptosis through complex multihormonal regulation during adult life. It is thus an attractive model to assess the in vivo function of some genes potentially involved in these mechanisms, either by silencing them or by partially repressing their expression. Furthermore, such alterations of gene expression have also been performed for more applied objectives such as the modification of milk composition for nutritional and technological purposes. This review will describe the experimental procedures used toward these aims and the results already obtained in this field. Some potential new targets will be suggested. PMID- 10819521 TI - The mammary fat pad. AB - The mammary fat pad is essential for development of the mammary epithelium, providing signals that mediate ductal morphogenesis and, probably, alveolar differentiation. The "cleared" fat pad is often used as a transplantation site. Considering the crucial role of the fat pad, its properties have received relatively little attention from researchers in the field. Some of the questions whose investigation is pertinent to understanding both normal mammary development and carcinogenesis are outlined in this commentary in the spirit of stimulating enquiry into this important subject. It is clear from a brief perusal of the available literature that until studies are specifically designed to clearly differentiate between functional effects of the fibrous and the adipose stroma, more substantive information about their differential effects on mammary development and tumorigenesis will not be forthcoming. PMID- 10819522 TI - The role of mammary stroma in modulating the proliferative response to ovarian hormones in the normal mammary gland. AB - Postnatal mammary gland development is highly dependent on the ovarian steroids, estrogen and progesterone. However, evidence from both in vitro and in vivo studies indicates that steroid-induced development occurs indirectly, requiring stromal cooperation in epithelial proliferation and morphogenesis. Stromal cells appear to influence epithelial cell behavior by secretion of growth factors and/or by altering the composition of the extracellular matrix in which epithelial cells reside. This review will discuss the requirement for stromal tissue in modulating proliferative responses to ovarian hormones during postnatal development and the potential role of the EGF, IGF, HGF and FGF3 growth factor families. Additionally, the roles of extracellular matrix proteins, including fibronectin, collagens and laminin, will be summarized. PMID- 10819523 TI - Roles of hepatocyte growth factor/scatter factor and transforming growth factor beta1 in mammary gland ductal morphogenesis. AB - Epithelial-mesenchymal interactions are responsible for the unique pattern of ductal branching morphogenesis characteristic of the mammary gland. To investigate the factors which control the elongation and branching of lactiferous ducts, we developed an in vitro model of ductal morphogenesis in which clonal mouse mammary epithelial cells (TAC-2 cells) are grown in collagen gels. In this experimental system, fibroblast conditioned medium (CM)3 stimulates the formation of extensively arborized tubules. The molecule responsible for this tubulogenic effect was identified as hepatocyte growth factor/scatter factor (HGF/SF). To determine whether HGF/SF plays a role in mammary gland morphogenesis in vivo, the expression of HGF/SF and its receptor, c-Met, were analyzed in the rat mammary gland during pregnancy, lactation, and involution. Levels of HGF/SF and c-Met transcripts were progressively reduced during pregnancy, were virtually undetectable during lactation, and increased again during involution. Collectively, these in vitro and in vivo findings suggest that HGF/SF is a paracrine mediator of mammary gland ductal morphogenesis. We subsequently investigated the effect of another multifunctional cytokine, namely TGF-beta1, on branching morphogenesis of TAC-2 cells. TGF-beta1 had a striking biphasic effect: whereas relatively high concentrations of this cytokine inhibited colony formation, lower concentrations stimulated extensive elongation and branching of epithelial cords. Taken together, these studies indicate that HGF/SF is a stromal derived paracrine mediator of mammary ductal morphogenesis, and that when present at low concentrations, TGF-beta1 can contribute to this process. PMID- 10819524 TI - Control of normal mammary epithelial phenotype by integrins. AB - Mammary epithelial cells contact a specialized extracellular matrix in vivo known as the basement membrane. Interactions with extracellular matrix are mediated through integrins. These cell surface receptors are involved with the formation of adhesion complexes, which link the extracellular matrix with the actin-based cytoskeleton, and are also associated with components of growth factor signaling pathways. Differentiation of breast epithelia into lactational cells requires appropriate hormones and integrin-mediated interactions with basement membrane. Integrins may regulate the ability of lactogenic hormones to trigger their intracellular signaling pathways. PMID- 10819525 TI - The potential influence of radiation-induced microenvironments in neoplastic progression. AB - Ionizing radiation is a complete carcinogen, able both to initiate and promote neoplastic progression and is a known carcinogen of human and murine mammary gland. Tissue response to radiation is a composite of genetic damage, cell death and induction of new gene expression patterns. Although DNA damage is believed to initiate carcinogenesis, the contribution of these other aspects of radiation response are beginning to be explored. Our studies demonstrate that radiation elicits rapid and persistent global alterations in the mammary gland microenvironment. We postulate that radiation-induced microenvironments may affect epithelial cells neoplastic transformation by altering their number or susceptibility. Alternatively, radiation induced microenvironments may exert a selective force on initiated cells and/or be conducive to progression. A key impetus for these studies is the possibility that blocking these events could be a strategy to interrupt neoplastic progression. PMID- 10819526 TI - Matrix metalloproteinases in remodeling of the normal and neoplastic mammary gland. AB - Alterations in mammary gland structure and function are associated with changes in the expression of members of the matrix metalloproteinase (MMP)3 family of enzymes. In this review, the evidence for a role for specific MMPs in mammary gland development and cellular differentiation, proliferation, and apoptosis is discussed. In addition, MMP expression is altered during the development and progression of preneoplastic and neoplastic breast lesions. The expression of MMP family members in human breast cancer is described, and studies with mouse model systems addressing the role of MMPS in the initiation, growth, invasion, and metastasis of breast neoplasms are reviewed. PMID- 10819527 TI - Altered integrin expression and the malignant phenotype: the contribution of multiple integrated integrin receptors. AB - The integrins are a family of cell surface adhesion receptors that mediate adhesion to either components of the extracellular matrix or to other cells. The beta1 family of integrins represent the major class of cell substrate receptors with specificities primarily for collagens, laminins, and fibronectins. The role of the integrin family of cell surface adhesion receptors in normal mammary gland morphogenesis and the contributions of altered integrin receptor expression to the invasive and metastatic phenotype have been the primary focus of our lab, as well as a number of other laboratories. The alpha2beta1 integrin is expressed at high levels by normal differentiated epithelial cells including those of the normal breast. Using breast cancer as a model, we evaluated changes in integrin expression in malignancy. We and other investigators made the key observation that alpha2beta1 integrin expression is decreased in adenocarcinoma of the breast in a manner that correlates with the stage of differentiation. Studies of other adenocarcinomas have yielded similar results. When the alpha2beta1 integrin was reexpressed in a poorly differentiated mammary carcinoma that expressed no detectable alpha2 integrin subunit, a dramatic reversion of malignant phenotype to a differentiated epithelial phenotype was observed, indicating a critical role for alpha2beta1 expression in mammary gland differentiation. Other laboratories using monoclonal antibodies to competitively inhibit alpha2beta1 integrin adhesion or oncogenic transformation using c-erb2 have confirmed the important role of that alpha2beta1 integrin in mammary gland morphogenesis. Re-expression of the alpha2beta1 integrin also results in upregulation of both the alpha6 and beta4 integrin subunits. To determine the contribution of enhanced alpha6 and beta4 integrin expression to the abrogation of the malignant phenotype by alpha2beta1 integrin expression, we have now separately re-expressed the human alpha6 or beta4 integrin subunit in the breast cancer model. PMID- 10819528 TI - Structural cues from the tissue microenvironment are essential determinants of the human mammary epithelial cell phenotype. AB - Historically, the study of normal human breast function and breast disorders has been significantly impaired by limitations inherent to available model systems. Recent improvements in human breast epithelial cell lines and three-dimensional (3-D)3 culture systems have contributed to the development of in vitro model systems that recapitulate differentiated epithelial cell phenotypes with remarkable fidelity. Molecular characterization of these human breast cell models has demonstrated that normal breast epithelial cell behavior is determined in part by the precise interplay that exists between a cell and its surrounding microenvironment. Recent functional studies of integrins in a human model system provide evidence to support the idea that the structural stability afforded by integrin-mediated cell-extracellular matrix interactions is an important determinant of normal cellular behavior, and that alterations in tissue structure can give rise to tumorigenic progression. PMID- 10819529 TI - The role of stroma in breast carcinoma growth in vivo. AB - The malignant progression of tumors is thought to be related to the expression of oncogenes and loss of expression of tumor suppressor gene. These factors are intrinsic to the cancer cells themselves. However, carcinomas are also infiltrated by host cells (fibroblasts, endothelial cells, inflammatory cells) and surrounded by an extracellular matrix which is extensively remodeled. The extracellular matrix components and infiltrating host cells provide a microenvironment that conditions both tumor progression and the metastatic process. Transplantation of human tumors into athymic nude mice has become an important experimental approach to study the biology of human cancers. The different models developed so far are beginning to elucidate the role of matrix molecules, growth factors and enzymes as well as fibroblasts in tumor progression. These animal models are likely to provide a useful tool to evaluate new antitumor treatments. PMID- 10819530 TI - Introduction: BRCA1 and BRCA2 in mammary gland development and tumorigenesis. PMID- 10819532 TI - Screening and clinical implications for BRCA1 and BRCA2 mutation carriers. AB - In this article, we review the history of testing for mutations in breast cancer susceptibility genes and discuss the current state of testing for mutations in BRCA1 and BRCA2 in different clinical settings including at-risk individuals and cancer patients. The risk of breast cancer. other associated malignancies and prognosis in carriers of these mutations are reviewed. A final section includes discussion of current recommendations for surveillance and the need for further research to identify environmental and genetic factors which modify the risk of developing cancer in mutation carriers. PMID- 10819531 TI - The genetics of inherited breast cancer. AB - The isolation of BRCA1 and BRCA2 has greatly increased our understanding of the genetics of inherited breast and ovarian cancer although the functions of these genes are not yet fully understood. We will discuss the current state of knowledge about the function of BRCA1 and BRCA2 and summarize the cancer risks in women carrying a BRCA1 or BRCA2 mutation. We review the evidence for gene-gene and gene-environment interaction in modifying that risk, and discuss the contribution of BRCA1 and BRCA2 and other high penetrance genes to both inherited and sporadic breast cancer. PMID- 10819533 TI - Expression of BRCA1 and BRCA2 in normal and neoplastic cells. AB - Current evidence strongly supports a role for the breast cancer susceptibility genes, BRCA1 and BRCA2, in both normal development and carcinogenesis. Valuable clues regarding the function of these genes have been garnered through studies of their patterns of expression. A central feature of the in vivo pattern of BRCA1 and BRCA2 expression is that each of these putative tumor suppressor genes is expressed at maximal levels in rapidly proliferating cells. This feature is consistent with in vitro observations that BRCA1 and BRCA2 are expressed in a cell cycle-dependent manner. This feature is also well illustrated during mammary gland development wherein the expression of BRCA1 and BRCA2 is induced in rapidly proliferating cellular compartments undergoing differentiation, such as terminal end buds during puberty and developing alveoli during pregnancy. Strikingly, the spatial and temporal patterns of BRCA1 and BRCA2 expression are virtually indistinguishable during embryonic development and in multiple adult tissues despite the fact that these genes are unrelated. These observations have contributed to the emerging hypothesis that these genes function in similar regulatory pathways. PMID- 10819534 TI - Functional domains of the BRCA1 and BRCA2 proteins. AB - The BRCA1 and BRCA2 genes encode large unrelated proteins that presumably function as tumor suppressors in normal epithelial cells of the breast. However, the primary amino acid sequences of these proteins provide few insights into the mechanisms by which BRCA1 and BRCA2 inhibit tumor development. Nevertheless, recent studies have uncovered many similarities in the biological properties of BRCA1 and BRCA2, raising the prospect that these proteins may function in a common pathway of tumor suppression and that inactivation of either gene may represent an equivalent step in the development of breast cancer. Several lines of evidence now suggest a role for BRCA1 and BRCA2 in the cellular response to DNA damage, possibly by virtue of their relationship with proteins required for the recombinational repair of double-strand DNA breaks. Accordingly, the loss of BRCA1 or BRCA2 function might accelerate tumor development by allowing cells to accumulate DNA lesions that are potentially oncogenic. PMID- 10819535 TI - Functional characterization of BRCA1 and BRCA2: clues from their interacting proteins. AB - The familial breast and ovarian cancer susceptibility genes, BRCA1 and BRCA2 have been the subject of extensive functional analysis studies since their cloning. Clues to their biological role in maintaining the genomic integrity were provided by studies that revealed their interaction with the recombination repair protein HsRad51. The first clue of an interaction between HsRad51 and BRCA1 came from the colocalization of the characteristic nuclear foci formed by these two proteins during S phase of the cell cycle. An interaction between murine Brca2 and MmRad51 was detected by the yeast two hybrid system. Utilizing the yeast two hybrid system and other techniques several other Brca1 and Brca2 interacting proteins have been identified like, BARD1, importin-alpha, BIPs, RNA polymerase II holoenzyme, BRAP2 etc. Recently, mutations suggesting a role as a tumor suppressor have been identified in the BARD1 gene in primary human tumors. The identification of molecules that interact with Brca1 and Brca2 has greatly enhanced our knowledge of how BRCA1 and BRCA2 may function as tumor suppressors. PMID- 10819536 TI - Localization of BRCA1 protein at the cellular level. AB - Based on its amino acid sequence and the existence of three nuclear localization signal (NLS)3 regions, BRCA1 is likely to be a cell cycle-dependent nuclear protein, regulated by cyclin-dependent kinases (cdk) and associated with nuclear proteins such as Rad51 and BARD1, involved in transcription regulation and participating in DNA replication checkpoints. However, many authors have also described a cytoplasmic expression pattern. Moreover, BRCA1 was present not only in a dot like pattern in the nucleus but also associated with a channel-like system of cytoplasm and endoplasmic reticulum invaginating into the nucleus. BRCA1 expression patterns can also be influenced by alternative splice variants and by cell cycle-dependent expression level and localization. Further ultrastructural and confocal studies using C-terminal antibodies, that do not react with C-terminal truncated form of BRCA1 should shed new light upon the exact localization of BRCA1. PMID- 10819538 TI - How proper protocol, materials selection, and communication yield aesthetic, functional outcomes and patient satisfaction in complex restorative cases. PMID- 10819539 TI - Paper alert. Cell biology. PMID- 10819537 TI - Developmental studies of Brca1 and Brca2 knock-out mice. AB - In humans, the inheritance of mutations in the breast cancer susceptibility genes BRCA1 and BRCA2 increases the risk of developing breast and ovarian cancer. To study their biological function and to create animal models for these cancer susceptibility genes, several strains of mice mutated in the homologous genes Brca1 and Brca2 have been generated by gene targeting. Analyses of these "knock out" mouse mutants have provided invaluable knowledge about the function of these genes. Brca1 and Brca2 null mutants are similar in phenotype: mutations in both genes result in embryonic lethality and the developing embryos show signs of a cellular proliferation defect associated with activation of the p53 pathway. The significance of this activation, as well as the role of these cancer susceptibility genes in DNA damage repair, is discussed. PMID- 10819540 TI - Web alert. Cell regulation. PMID- 10819541 TI - BMP, Wnt and Hedgehog signals: how far can they go? AB - Wnt, Hedgehog and bone morphogenetic proteins function as either short-range or long-range signaling molecules depending on the tissue in which they are expressed. In the past year, filapodia-like cytoplasmic extensions, cell-surface proteogylcans and/or extracellular binding proteins have been identified that may enable these molecules to signal at a distance. Furthermore, recent studies suggest that variations in the signaling range of these molecules may be due to tissue-specific differences in intracellular processing or tissue-restricted expression of binding proteins. PMID- 10819542 TI - Apoptosis and cell division. PMID- 10819543 TI - [Secondary prevention of coronary artery disease: interaction between the hospital and outpatient clinic in or der to form successful, low-cost strategies]. PMID- 10819544 TI - Peristomal lichen sclerosus affecting colostomy sites. PMID- 10819545 TI - COL3A1 mutation leading to acrogeria (Gottron Type) PMID- 10819546 TI - Zosteriform relapse of B-cell lymphoma. PMID- 10819547 TI - Zosteriform metastasis of colonic carcinoma. PMID- 10819548 TI - Subcutaneous T-cell lymphoma in a child with eosinophilia. PMID- 10819549 TI - Melanocytes in squamous cell carcinoma of the skin. PMID- 10819550 TI - Ultraviolet-induced generalized acquired dermal melanocytosis with numerous melanophages. PMID- 10819551 TI - Perianal ulceration: a rare manifestation of tuberculosis. PMID- 10819552 TI - Hepatitis C virus: a common triggering factor for both nodular vasculitis and Sjogren's syndrome? PMID- 10819553 TI - Radiotherapy and erythema nodosum. PMID- 10819554 TI - Interferon alpha 2b for treatment of complex cutaneous haemangiomas of infancy: a reduced dosage schedule. PMID- 10819555 TI - A case of toxic epidermal necrolysis treated with intravenous immunoglobin. PMID- 10819556 TI - Angiolymphoid hyperplasia with eosinophilia successfully treated with the flash lamp pulsed-dye laser. PMID- 10819557 TI - A caution about surgical treatment for facial blushing. PMID- 10819558 TI - Staphylococcal toxins and the scalded skin syndrome. PMID- 10819559 TI - Lip papillomatosis in immunodeficiency: therapy with imiquimod. PMID- 10819560 TI - Crusted scabies in a girl with epidermolysis bullosa simplex. PMID- 10819561 TI - Isoretinoin associated with craving for cigarettes. PMID- 10819562 TI - On bacterial tactic response times and latencies. PMID- 10819563 TI - Transfusion-related acute lung injury. PMID- 10819564 TI - Non-neoplastic mortality of European workers who produce man made vitreous fibres. PMID- 10819565 TI - Karo jisatsu (suicide from overwork): a spreading occupational threat. PMID- 10819566 TI - The industrial athlete. PMID- 10819567 TI - CQ sources/bibliography. PMID- 10819568 TI - Electroporation-enhanced gene delivery in mammary tumors. AB - Electroporation was applied to enhance gene transfer into subcutaneous MC2 murine breast tumors. Cultured MC2 cells were also transfected by electroporation or by cationic liposomes in the presence of serum using pSV-luc plasmids. Electroporation parameters and liposome formulation were optimized to achieve the highest relative levels of transfection. An electric field threshold for successful electrotransfection in cultured cells appeared around 800-900 V/cm. The liposomes used contained the cationic lipid dioleoyl-3-trimethylammonium propane (DOTAP). Multilamellar vesicles (MLV) had a 10-fold advantage over small unilamellar vesicles (SUV) in cell culture transfection. For in vivo gene delivery, the plasmids were injected either alone, or in complex with MLV or SUV DOTAP liposomes. A series of six electric pulses 1 ms long were applied across tumors, using caliper electrodes on the skin surface. Electric field strengths ranged from 400-2300 V/cm. Luciferase expression was approximately two orders of magnitude higher than controls in tumors treated with pulses > or =800 V/cm. Differences between enhanced relative levels of transfection using uncomplexed plasmid and lipoplexes were not statistically significant. Distribution of DNA into tumor tissues was monitored by fluorescence in situ PCR. The highest numbers of fluorescent cells were found in tumors electroporated following the injection of plasmid. The significant transfection improvement shows that in vivo electroporation is a powerful tool for local gene delivery to tumors. PMID- 10819569 TI - Hepatocyte-targeted in vivo gene expression by intravenous injection of plasmid DNA complexed with synthetic multi-functional gene delivery system. AB - To achieve hepatocyte-targeted in vivo gene expression, a carrier that controls both the tissue and intracellular distribution of DNA was designed and synthesized. A cationic polymer, poly(L-ornithine) (pOrn), was modified first with galactose, then with a fusigenic peptide (mHA2) to obtain Gal-pOrn-mHA2. When applied with Gal-pOrn-mHA2 to asialoglycoprotein receptor-positive cells, fluorescein-labeled DNA showed a diffuse profile, suggesting the release of DNA from endosomes and/or lysosomes by the carrier. Then the biodistribution and gene expression after intravenous injection of DNA complexes (10 microg DNA per mouse) were examined. After injection of [32P]DNA/Gal-pOrn-mHA2, about 60% of the radioactivity was recovered in the liver, mostly in parenchymal cells. A large amount (81 ng/g tissue) of transgene product (luciferase) was detected in the liver of mice injected with DNA/Gal-pOm-mHA2, which was 280-fold greater than that obtained with DNA/DOTMA:Chol liposomes (50 microg DNA). Prior administration of galactosylated albumin reduced the gene expression to 1/100, indicating the asialoglycoprotein receptor-mediated gene transfer in liver parenchymal cells, ie hepatocytes. The luciferase activity in hepatocytes contributed more than 95% of the total activity in all the tissues examined. Thus, hepatocyte-targeted in vivo gene expression was achieved by the intravenous injection of DNA complex with the multifunctional gene carrier. PMID- 10819570 TI - Insulation from viral transcriptional regulatory elements improves inducible transgene expression from adenovirus vectors in vitro and in vivo. AB - Recombinant adenoviruses (Ad) are attractive vectors for gene transfer in vitro and in vivo. However, the widely used E1-deleted vectors as well as newer generation vectors contain viral sequences, including transcriptional elements for viral gene expression. These viral regulatory elements can interfere with heterologous promoters used to drive transgene expression and may impair tissue specific or inducible transgene expression. This study demonstrates that the activity of a metal-inducible promoter is affected by Ad sequences both upstream and downstream of the transgene cassette in both orientations. Interference with expression from the heterologous promoter was particularly strong by viral regulatory elements located within Ad sequences nucleotides 1-341. This region is present in all recombinant Ad vectors, including helper-dependent vectors. An insulator element derived from the chicken gamma-globin locus (HS-4) was employed to shield the inducible promoter from viral enhancers as tested after gene transfer with first-generation Ad vectors in vitro and in vivo. Optimal shielding was obtained when the transgene expression cassette was flanked on both sides by HS-4 elements, except for when the HS-4 element was placed in 3'-->5' orientation in front of the promoter. The insulators reduced basal expression to barely detectable levels in the non-induced stage, and allowed for induction factors of approximately 40 and approximately 230 in vitro and in vivo, respectively. Induction ratios from Ad vectors without insulators were approximately 40-fold lower in vitro and approximately 15-fold lower in vivo. This study proves the potential of insulators to improve inducible or tissue-specific gene expression from adenovirus vectors, which is important for studying gene functions as well as for gene therapy approaches. Furthermore, our data show that insulators exert enhancer-blocking effects in episomal DNA. PMID- 10819571 TI - Pseudotyped human lentiviral vector-mediated gene transfer to airway epithelia in vivo. AB - We used a replication defective human lentiviral (HIV) vector encoding the lacZ cDNA and pseudotyped with the vesicular stomatitis virus (VSV) glycoprotein (G) to evaluate the utility of this vector system in airway epithelia. In initial studies, apical application of vector to polarized well differentiated human airway epithelial cell cultures produced minimal levels of transgene expression whereas basolateral application of vector enhanced levels of transduction approximately 30-fold. Direct in vivo delivery of HIV vectors to the nasal epithelium and tracheas of mice failed to mediate gene transfer, but injury with sulfur dioxide (SO2) before vector delivery enhanced gene transfer efficiency to the nasal epithelium of both mice and rats. SO2 injury also enhanced HIV vector mediated gene transfer to the tracheas of rodents. These data suggest that SO2 injury increases access of vector to basal cells and/or the basolateral membrane of airway surface epithelial cells. Quantification of gene transfer efficiency in murine tracheas demonstrated that transduction was more efficient when vector was delivered on the day of exposure (7.0%, n = 4) than when vector was delivered on the day after SO2 exposure (1.7%, n = 4). PMID- 10819573 TI - The use of oligonucleotide directed cleavage of DNA and homologous recombination in the production of large recombinant adenoviral vectors. AB - The genetic manipulation of large plasmid DNA often requires the fortuitous presence of convenient restriction enzyme sites. For large plasmids, such as those containing full length recombinant adenovirus, it is desirable to direct the cloning or sequence alterations without having to depend on such convenient restriction sites. We report a general and efficient method to modify or clone large covalently closed circular DNA molecules at any predetermined sequence. This procedure involves two main steps. First, supercoiled DNA is hybridized to a short pre-selected synthetic oligonucleotide to form a D-loop. This hybrid is then linearized in vitro at that target site by digestion with S1 nuclease. Second, D-loop/S1 linearized DNA is transformed into E. coli with a second linear DNA fragment carrying a foreign gene flanked by sequences homologous to the target site. In vivo recombination results in the desired recombinant construct. We demonstrate the use of this method by cloning the SV40 origin of replication into the E3 transcription unit of human adenovirus type 5. PMID- 10819572 TI - Gene transfer of virally encoded chemokine antagonists vMIP-II and MC148 prolongs cardiac allograft survival and inhibits donor-specific immunity. AB - Introducing immunomodulatory molecules into allografts by gene transfer may avoid the side-effects of systemic immunosuppression. vMIP-II and MC148 are two recently identified chemokine homologues encoded by human herpes virus 8 and Molluscum contagiosum, respectively, that have antagonistic activities against multiple different CC and CXC chemokine receptors. We hypothesized that introduction of these molecules into cardiac allografts may block leukocyte infiltration into the grafts and prolong survival. Vascularized and nonvascularized cardiac allografts in mice were performed and plasmid DNA encoding vMIP-II, MC148 and/or vIL-10 was transferred into the allograft at the time of transplantation. Gene transfer of either vMIP-II or MC148 into cardiac allografts markedly prolonged graft survival. Combining gene transfer of either one of these chemokine antagonists with vIL-10 gene transfer, which has a mechanistically different immunosuppressive action, further enhanced graft survival. vMIP-II and MC148 gene transfer both resulted in a marked decrease of donor-specific cytotoxic T lymphocytes (CTL) infiltrating the grafts and inhibited alloantibody production. These results demonstrate that plasmid mediated gene transfer of virally encoded chemokine antagonists vMIP-II and MC148 can block donor-specific lymphocyte immunity within cardiac allografts and prolong graft survival. This is a new mechanistic approach to analyze, treat, and prevent graft rejection. Delivery of these or related molecules by gene transfer or conventional pharmacologic means may represent a novel therapeutic modality for alloactivation. PMID- 10819574 TI - Effective repeat administration with adenovirus vectors to the muscle. AB - Effective repeat administration of adenovirus vectors following intranasal or intravenous delivery is hindered by a strong neutralizing antibody response to the vector. Intramuscular administration of adenovirus vectors elicited a neutralizing antibody response that peaked between 14 and 21 days after infection. However, effective repeat intramuscular administration of adenovirus vectors was not hindered by the presence of neutralizing antibodies in the serum. Surprisingly, beta-galactosidase expression in the skeletal muscle of immunized mice was equivalent to that observed in control mice. As expected, these serum neutralizing antibodies effectively blocked repeat administration of adenovirus vectors when delivered via the intravenous route. These results were observed in both C57BL/6 and Balb/c mice and thus do not appear to be strain specific. Successful repeat administration of adenovirus vectors to skeletal muscle has significant implications for the use of adenovirus vectors clinically and for increasing the safety and efficacy of adenovirus vector gene delivery. PMID- 10819575 TI - Gene transfer into stimulated and unstimulated T lymphocytes by HIV-1-derived lentiviral vectors. AB - Genetic modification of T lymphocytes holds great potential for treatments of cancer, T cell disorders and AIDS. While in the past recombinant murine retroviruses were the vectors of choice for gene delivery to T cells, vectors based on lentiviruses can provide additional benefits. Here, we show that VSV-G pseudotyped HIV 1 vector particles delivering the enhanced green fluorescent protein (EGFP) efficiently transduce human T lymphocytes. Transduction efficiency was optimal when infection included centrifugation of cells with concentrated vector supernatant in the presence of Polybrene. In contrast to previous reports describing murine retrovirus-mediated gene transfer to T lymphocytes, fibronectin did not improve the transduction efficiency of the VSVG-pseudotyped HIV-1 particles. Similar gene transfer efficiencies were observed following stimulation of cells with PHA/IL-2 or anti-CD3i/CD28i antibodies, although greater transgene expression was observed in the latter case. Interestingly, production of vectors in the absence of the accessory proteins Vif, Vpr, Vpu and Nef was accompanied by a 50% decrease in transduction efficiency in activated T cells. Transduction of T cells that were not stimulated before infection was achieved. No transduction of non-prestimulated cells was observed with a GAL V-pseudotyped murine retroviral vector. The requirement for accessory proteins in non-prestimulated cells was more pronounced. Our results have implications for lentiviral vector targeting of other cells of the hematopoietic system including stem cells. PMID- 10819576 TI - Anti-idiotypic DNA vaccines for lymphoma immunotherapy require the presence of both variable region genes for tumor protection. AB - Vaccination with immunogenic formulations of lymphoma-derived immunoglobulin can elicit strong anti-idiotypic immune responses which have proved effective in murine B cell tumor challenge experiments and suggested possible benefits in recent human clinical trials. Naked plasmid DNA vaccines encoding the Id determinants as scFv fragments provide the most promising alternative to protein immunization. With this approach the addition of an immunogenic domain linked to the scFv has proved essential for the induction of a protective immune response. In this study we have produced a scFv gene construct linked to the CH3 exon of the human IgG1 constant region and tested its efficacy in inducing protective immunity against the mouse BCL1 lymphoma. We have also generated a second construct in which the BCL1 VL gene was deleted to investigate whether the VH region domain contains sufficient antigenic determinants for a protective immune response. Both constructs induced anti-idiotypic antibodies that specifically reacted with the BCL1 IgM protein in ELISA and with BCL1 tumor cells in flow cytometry assays. Protection against tumor challenge was fully achieved with the complete scFv construct whereas immunization with the construct lacking the VL gene resulted in only a slight prolongation of the survival. We therefore conclude that a plasmid DNA vaccine containing the VH and VL genes of the lymphoma Ig linked to the human IgG1 CH3 exon is highly effective in inducing a protective immune response in the BCL1 model. We also demonstrated that VH gene immunization can induce strong anti-idiotypic antibody responses. PMID- 10819577 TI - A murine tracheal culture system to investigate parameters affecting gene therapy for cystic fibrosis. AB - Cystic fibrosis (CF) is a life-threatening condition caused by mutations in the cystic fibrosis transmembrane conductance regulator gene (CFTR). Delivery of the CFTR gene to the airways offers a potential treatment for CF but requires improvement in efficiency to obtain clinical benefit. We have developed a murine tracheal culture system that maintains tissue integrity as judged by normal histological appearance, high transepithelial resistance and electrophysiological responses similar to fresh tissue. This ex vivo system allows precise control of gene delivery parameters to a structure that retains the in vivo cellular architecture. We have demonstrated correction of CFTR-dependent Cl- secretion following ex vivo delivery of the CFTR gene to tracheas from CF null mice. We have used this system to examine parameters affecting liposome-mediated gene delivery to the upper airway such as plasmid dose. We have also found that a contact time of 1 min for the transfection mixture is sufficient to achieve significant DNA binding and maximal reporter gene expression. PMID- 10819578 TI - Telomerase allows the immortalization of T antigen-positive DMD myoblasts: a new source of cells for gene transfer application. AB - The limited proliferative capacity of DMD myoblasts severely limits their ability to be genetically modified and used for myoblast transplantation. Transformation by SV40 large T antigen (Tag) delays senescence of mouse and human myoblasts but fails to immortalize these cells. The cells ceased to proliferate and entered into crisis. Reconstitution of telomerase activity has been shown sufficient to enable different types of transformed cells to escape crisis. DMD myoblasts, previously transformed by Tag, were therefore infected with a telomerase retrovirus. The expression of telomerase was sufficient to allow DMD-Tag myoblasts to escape crisis. The telomerase-positive transformed myoblasts continued to divide for more than 55 doublings while Tag myoblasts stopped proliferating after 35 doublings. These cells are able to fuse and to differentiate normally. The average telomere length of these telomerase-positive DMD-Tag myoblasts seems to continue to elongate. Thus, transiently genetically modified myoblasts could constitute an important pool of DMD myoblasts for autologous transplantation in DMD patients. PMID- 10819579 TI - Efficacy of cancer gene therapy in aging: adenocarcinoma cells engineered to release IL-2 are rejected but do not induce tumor specific immune memory in old mice. AB - Numerous studies have demonstrated the efficacy of cytokine gene-engineered tumor cells to induce tumor rejection and specific memory acquisition into syngeneic immunocompetent mice by activation of host-dependent antitumor responses. A progressive immune dysfunction, mainly involving thymus-dependent specific immunity, occurs during aging. In this study we evaluated whether the injection of IL-2 gene-transfected tumor cells in old mice causes an immune activation which results in tumor rejection and induction of specific immune memory as occurs in young animals. Young and old mice were inoculated with syngeneic parental mammary adenocarcinoma cells (TS/A p.c.) or with TS/A cells engineered to release IL-2 (TS/A-IL2). Three clones of TS/A-IL-2 cells were used producing low (30 U, B1.30), intermediate (3600 U, B6.3600), or high (6000 U, B4.6000) IL 2. While the B1.30 clone grew in 100% of mice, the B6.3600 and B4.6000 clones were promptly rejected in both young and old animals. In young mice, rejection was associated with a large neutrophil and macrophage infiltration, with a minor number of CD4+ and CD8+ lymphocytes. In old mice, neutrophils and macrophages were the main cells involved in tumor rejection whereas both CD4+ and CD8+ lymphocytes were scarcely present in tumoral infiltrate. A lower number of apoptotic tumor cells was found in TS/A-IL2-challenged old mice in comparison with young animals. To test whether the injection of TS/A-IL2 cells induced a specific immune memory, mice with no tumors after the challenge with B6.3600 and B4.6000 clones received a lethal challenge of TS/A p.c. 90% and 30% of young mice previously injected with B4.6000 or B6.3600 clones, respectively, rejected TS/A p.c. In old mice, B4.6000 cells did not confer protection, whereas only 10% of mice which received B6.3600 cells were able to reject TS/A p.c. Neither the graft of a young thymus or the adoptive transfer of young T lymphocytes to old mice induced specific immune memory for TS/A p.c. in old animals. These data suggest the necessity to refine antitumor vaccination procedures in aging. PMID- 10819580 TI - German-Austrian guidelines for antiretroviral therapy in HIV infection. June 1999. Deutsche AIDS-Gesellschaft (DAIG). Osterreichische AIDS-Gesellschaft (OAG). PMID- 10819581 TI - Notable and anniversary dates in biochemistry for 2000. PMID- 10819582 TI - [Abstracts of work presented at the 49th Pharmacology Seminar held in Bratislava (8-10 September 1999)]. PMID- 10819583 TI - 5th International Workshop on Computer Assisted Surgery and Rapid Prototyping in Medicine. Erlangen, Germany, October 14-16, 1999. Abstracts. PMID- 10819584 TI - Proceedings of the 3rd International Conference on Human Herpesviruses 6, 7 and 8. Clearwater Beach, Florida, USA. May 13-15, 1999. PMID- 10819585 TI - Historical review of the combined oral contraceptive pill and injectable progestogens. PMID- 10819586 TI - Development of contraceptive implants. PMID- 10819587 TI - The current status of intra-uterine contraceptive devices and systems. PMID- 10819588 TI - Contraception for the older woman-a century of change. PMID- 10819589 TI - Termination of pregnancy: past, present and future. PMID- 10819590 TI - The global challenges for family planning in the 21st century. PMID- 10819591 TI - Faculty of Family Planning and Reproductive Health Care. Royal College of Obstetricians and Gynaecologists. First prescription of combined oral contraception: recommendations for clinical practice. PMID- 10819592 TI - Female genital mutilation; its implications for reproductive health. An overview. PMID- 10819593 TI - Contraceptives for the 21st century. PMID- 10819594 TI - [Usefulness of 99mTc-DTPA cysternoscintigraphy in CSF fistula in the sphenoid region]. PMID- 10819595 TI - [Informed consent in radioisotopic synoviorthesis]. PMID- 10819596 TI - Dimetallic hydrolases and their models. AB - Recent mechanistic studies of Fe3+/M2+ purple acid phosphatase present conflicting pictures about the roles of the metals. Recent model studies with Co3+/Co3+ and Fe3+/Fe3+ complexes have supported each conflicting mechanism for phosphate hydrolysis. With dizinc and trizinc complexes, the competitive binding of hydroxyl and phosphate substrates has been demonstrated. PMID- 10819597 TI - Great leaps forward: translesion synthesis gets unstalled. PMID- 10819598 TI - Changes in intercellular junctions during apoptosis precede nuclear condensation or phosphatidylserine exposure on the cell surface. PMID- 10819599 TI - The apoptotic regulatory gene, BCL10, is mutated in sporadic mismatch repair deficient colorectal cancers. PMID- 10819600 TI - Frameshift mutations in Fas, Apaf-1, and Bcl-10 in gastro-intestinal cancer of the microsatellite mutator phenotype. PMID- 10819601 TI - Flaws within travel medicine. PMID- 10819603 TI - Credentialing of travel cities. PMID- 10819602 TI - Weight-based units reporting of anti-Vi antibody concentrations. PMID- 10819604 TI - Delayed primary attack of vivax malaria. PMID- 10819605 TI - Papular fixed drug eruption mimicking folliculitis due to acetominophen. PMID- 10819606 TI - Gingival hyperplasia in psoriasis patients treated with cyclosporin. PMID- 10819607 TI - Verrucous cutaneous sarcoidosis. PMID- 10819608 TI - The discovery of the ophthalmoscope by Herman von Helmholtz (1824-1894). PMID- 10819609 TI - J.F. Dieffenbach: treatment of congenital squint by section of the medial rectus. PMID- 10819610 TI - What speaks in favor of a complete correction of renal anemia? Nephrology Workshop. Berlin, Germany, October 10, 1998. PMID- 10819611 TI - P1 blood group and haemolytic uraemic syndrome. PMID- 10819612 TI - Site of subcutaneous desferrioxamine injection is the initial site of post transfusion urticaria in multi-transfused beta-thalassaemia major patients. PMID- 10819613 TI - Artefactual elevation of the automated white cell count in the context of a bone marrow aspirate analysis. PMID- 10819614 TI - Stability of prothrombin time and the impact of container material. PMID- 10819615 TI - Meera Khan dedication issue. PMID- 10819616 TI - Vitamin A supplementation in early infancy. PMID- 10819617 TI - Atrial myxoma identified by transesophageal echocardiography in a patient with NAME syndrome. PMID- 10819618 TI - Verrucous varicella zoster virus lesions associated with acquired immunodeficiency syndrome. PMID- 10819619 TI - Basal cell carcinoma associated with chronic venous leg ulcer. PMID- 10819620 TI - Corticosteroid-resistant erosive oral lichen planus successfully treated with topical cyclosporine therapy. PMID- 10819621 TI - Ambulatory impedance cardiography: new possibilities. PMID- 10819622 TI - Direct urease test on BACTEC blood cultures: early presumptive diagnosis of brucellosis in an area of endemicity. PMID- 10819623 TI - Generation of oxoiron (IV) tetramesitylporphyrin pi-cation radical complexes by m CPBA oxidation of ferric tetramesitylporphyrin derivatives in butyronitrile at - 78 degrees C. Evidence for the formation of six-coordinate oxoiron (IV) tetramesitylporphyrin pi-cation radical complexes FeIV = O(tmp*)X (X = Cl-, Br-), by Mossbauer and X-ray absorption spectroscopy. AB - The generation of six-coordinate oxoiron (IV) tetramesitylporphyrin pi-caption radical complexes by m-CPBA (meta-chloroperbenzoic acid) oxidation of ferric tetramesitylporphyrin derivatives in butyronitrile at - 78 degrees C was investigated. UV-Vis and EPR spectroscopies indicate that the axial ligand present in the ferric starting derivatives is retained in the high-valent iron complexes. Indirect evidence for the formation of six-coordinate oxoiron (IV) tetramesitylporphyrin complexes FeIV = O(tmp*)X (X=Cl-, Br-) by m-CPBA oxidation of FeX(tmp) (X=Cl-, Br-) in butyronitrile at - 78 degrees C was also obtained by Mossbauer spectroscopy. Direct confirmation of the presence of a halide ion as second axial ligand of iron in these high-valent iron species was obtained by X ray absorption spectroscopy. The EXAFS spectra of the samples obtained by m-CPBA oxidation of FeX(tmp) (X=Cl-, Br-) were refined using two different coordination models including both four porphyrinato-nitrogens and the axial oxo group. The two models include (model I) or exclude (model II) the axial halogen. The statistical tests indicate the presence of a halide ion as second axial ligand of iron in both derivatives. The refinements led to the following bond distances: FeIV=O(tmp*)Cl(3):Fe-O=1.66(1),Fe-Cl=2.39(2) and Fe-Np=1.99(1) A;FeIV=O(tmp*)Br(4):Fe-O=1.65(1),Fe-Br=2.93(2), Fe-Np=2.02(1) A. The lengthening of the Fe-X(X=Cl-, Br-) distances relative to those occurring in the ferric precursor porphyrins is, most probably, related to the strong trans influence of the oxoiron(IV) fragment present in 3 or 4. PMID- 10819624 TI - Evaluation of intramolecular equilibria in complexes formed between substituted imidazole ligands and nickel (II), copper (II) or zinc (II). AB - The metal ion-binding properties of imidazole-4-acetate (ImA-), 4(5) aminoimidazole-5(4)-carboxamide (AImC), 2,2'biimidazole(BiIm) (I. Torok et al., J. Inorg. Biochem. 71 (1998) 7-14), and bis (imidazol-2-yl)methane(BiImM) (K. Varnagy et al., J. Chem. Soc., Dalton Trans. (1994) 2939-2945) have been evaluated by using the recently published stability constants and by applying the recently established log K(ML)M versus pK(HL)H straight-line plots (L. E. Kapinos et al., Inorg. Chim. Acta 280 (1998) 50-56) which hold for simple imidazole-type ligands. The indicated analysis regarding the intramolecular equilibrium between a monodentatally imidazole-nitrogen-coordinated (open) species and a chelated isomer provides helpful insights, e.g., the formation degree of chelates is more favored if six-membered rings can be formed, as in the case with M(BiImM)2+ compared to M(BiIm)2+, though in both instances the formation degree of the chelates is large. The formation degree of chelates in the M(ImA)+ complexes increases in the series Zn(ImA)+ (87%)G shows a severe phenotype. PMID- 10819641 TI - Alkaptonuria in Italy: polymorphic haplotype background, mutational profile, and description of four novel mutations in the homogentisate 1,2-dioxygenase gene. PMID- 10819639 TI - Three novel SALL1 mutations extend the mutational spectrum in Townes-Brocks syndrome. PMID- 10819642 TI - Rough skin, brittle hair, and photosensitivity: a mild phenotypic variant of trichothiodystrophy. PMID- 10819643 TI - The first description of lethal pterygium syndrome with facial clefting(Bartsocas Papas syndrome) in 1600. PMID- 10819644 TI - Sex reversal and diaphragmatic hernia in phenotypicaly female sibs with normal XY chromosomes. PMID- 10819645 TI - Clinical variability of Stickler syndrome with a COL2A1 haploinsufficiency mutation: implications for genetic counselling. PMID- 10819646 TI - [Diagnosis of autoimmune hepatitis]. PMID- 10819647 TI - [Critical remarks on the state of research in Germany]. PMID- 10819648 TI - Hepatic arterial infusion of chemotherapy for metastatic colorectal cancer. PMID- 10819649 TI - Hepatic arterial infusion of chemotherapy for metastatic colorectal cancer. PMID- 10819651 TI - Hepatic arterial infusion of chemotherapy for metastatic colorectal cancer. PMID- 10819650 TI - Hepatic arterial infusion of chemotherapy for metastatic colorectal cancer. PMID- 10819652 TI - Volume of procedures at transplantation centers and mortality after liver transplantation. PMID- 10819653 TI - Volume of procedures at transplantation centers and mortality after liver transplantation. PMID- 10819654 TI - Volume of procedures at transplantation centers and mortality after liver transplantation. PMID- 10819655 TI - Firearms and suicide. PMID- 10819656 TI - Congenital autoimmune diabetes mellitus. PMID- 10819657 TI - Patients 65 years of age or older in cancer-treatment trials. PMID- 10819658 TI - Patients 65 years of age or older in cancer-treatment trials. PMID- 10819659 TI - More on chewing gum. PMID- 10819660 TI - Anabolic-androgenic steroids as a gateway to opioid dependence. PMID- 10819661 TI - California targets GM-trial vandals with new legislation. PMID- 10819662 TI - Bid to relax rules on tissue transport runs into opposition. PMID- 10819663 TI - Politicians seek to block human-gene patents in Europe. PMID- 10819664 TI - Oxidant stress in hemodialysis. PMID- 10819665 TI - Fungemia in neonates: report of 80 cases from seven University hospitals. PMID- 10819666 TI - Insulin infusions in extremely low birth weight infants. PMID- 10819667 TI - Sleep location and suffocation: how good is the evidence? PMID- 10819668 TI - Sleep location and suffocation: how good is the evidence? PMID- 10819669 TI - Sleep location and suffocation: how good is the evidence? PMID- 10819671 TI - Multiethnic families: an underrecognized influence on health statistics. PMID- 10819670 TI - Theophylline-induced convulsions in children with epilepsy. PMID- 10819672 TI - Ultrasonography detects appendicular mucocele in cystic fibrosis patients suffering recurrent abdominal pain. PMID- 10819674 TI - [Peer review of scientific papers]. PMID- 10819675 TI - [Dentistry, a liberal profession--dream or reality?]. PMID- 10819676 TI - [Lasers in surgery--2]. PMID- 10819673 TI - [Negative feedback loop of clock oscillating genes]. PMID- 10819678 TI - [We repress emergency situations. Interview by Peter Jager]. PMID- 10819677 TI - [Anonymous (after Gerard Wigmana): the tooth-puller and a woman patient behind a balustrade. The BonaDent Collection]. PMID- 10819679 TI - [The Bone Symposium Bern, 11-12 December 1999]. PMID- 10819680 TI - [A report on the 4th Basel Materials Science Symposium of 3-4 December 1999]. PMID- 10819681 TI - [Laser therapy in the dental office--1]. PMID- 10819682 TI - Field of the future: neurogenic disease. PMID- 10819683 TI - British Thoracic Society Winter Meeting 1999. PMID- 10819684 TI - Mechanism of osteoporosis in patients with cystic fibrosis. PMID- 10819685 TI - Spirometry in primary care. PMID- 10819686 TI - Asthma guidelines. PMID- 10819687 TI - Respiratory questionnaires in COPD. PMID- 10819688 TI - Athletes and fenoterol. PMID- 10819689 TI - Asthma education. PMID- 10819690 TI - [Some data concerning the clinical investigation of birds of prey]. PMID- 10819691 TI - [The introduction of cesarean section into veterinary practice]. PMID- 10819692 TI - [Veterinary diseases, a veterinary problem?]. PMID- 10819693 TI - [Professor Nikolai Alekseevich Karpov (birth centenary)]. PMID- 10819694 TI - International Congress on Viral Hepatitis A and B: Experience in Education and Prevention. Warsaw, Poland, 18-21 October 1998. Proceedings. PMID- 10819695 TI - Antibiotic Resistance. Proceedings of the 11th Internordic Symposium of the Nordic Committee for Veterinary Scientific Cooperation (NKVet). Sundvolden, Norway, 7-8 November 1997. PMID- 10819696 TI - Proceedings of the 2nd International Workshop on Molecular Pathogenesis of Marek's Disease. Slovak Republic, August 8-11, 1998. PMID- 10819697 TI - Is there a parallel relation between the increased incidence of obesity and asthma? PMID- 10819698 TI - Automatico-voluntary dissociation in Angelman syndrome. PMID- 10819699 TI - Cost-effectiveness of cholesterol-lowering therapies according to selected patient characteristics. AB - BACKGROUND: The National Cholesterol Education Program Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel II) recommends treatment guidelines based on cholesterol level and number of risk factors. OBJECTIVE: To evaluate how the cost-effectiveness ratios of cholesterol-lowering therapies vary according to different risk factors. DESIGN: Cost-effectiveness analysis. DATA SOURCES: Published data. TARGET POPULATION: Women and men 35 to 84 years of age with low-density lipoprotein cholesterol levels of 4.1 mmol/L or greater (> or =160 mg/dL), divided into 240 risk subgroups according to age, sex, and the presence or absence of four coronary heart disease risk factors (smoking status, blood pressure, low-density lipoprotein cholesterol level, and high-density lipoprotein cholesterol level). TIME HORIZON: 30 years. PERSPECTIVE: Societal. INTERVENTIONS: Step I diet, statin therapy, and no preventive treatment for primary and secondary prevention. OUTCOME MEASURES: Incremental cost-effectiveness ratios. RESULTS OF BASE-CASE ANALYSIS: Incremental cost-effectiveness ratios for primary prevention with step I diet ranged from $1900 per quality-adjusted life-year (QALY) gained to $500000 per QALY depending on risk subgroup characteristics. Primary prevention with a statin compared with diet therapy was $54000 per QALY to $1400000 per QALY. Secondary prevention with a statin cost less than $50000 per QALY for all risk subgroups. RESULTS OF SENSITIVITY ANALYSIS: The inclusion of niacin as a primary prevention option resulted in much less favorable incremental cost-effectiveness ratios for primary prevention with a statin (>$500000 per QALY). CONCLUSIONS: Cost-effectiveness of treatment strategies varies significantly when adjusted for age, sex, and the presence or absence of additional risk factors. Primary prevention with a step I diet seems to be cost-effective for most risk subgroups but may not be cost-effective for otherwise healthy young women. Primary prevention with a statin may not be cost-effective for younger men and women with few risk factors, given the option of secondary prevention and of primary prevention in older age ranges. Secondary prevention with a statin seems to be cost-effective for all risk subgroups and is cost-saving in some high-risk subgroups. PMID- 10819700 TI - Cost-effectiveness of 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor therapy in older patients with myocardial infarction. AB - BACKGROUND: 3-Hydroxy-3-methylglutaryl coenzyme A reductase inhibitor (statin) therapy has proven efficacy in reducing the rate of coronary and cerebrovascular events in patients 75 years of age or younger with a history of myocardial infarction. However, in patients older than 75 years of age, the efficacy and potential cost-effectiveness of statins are unknown. OBJECTIVE: To estimate the incremental cost-effectiveness of statin therapy compared with usual care in patients 75 to 84 years of age with previous myocardial infarction. DESIGN: Cost effectiveness analysis. DATA SOURCES: Published data from cohort studies. TARGET POPULATION: Patients 75 to 84 years of age with a history of myocardial infarction. TIME HORIZON: Lifetime. PERSPECTIVE: Societal. INTERVENTION: Statin therapy. OUTCOME MEASURES: Life expectancy, quality-adjusted life expectancy, and incremental cost-effectiveness. RESULTS OF BASE-CASE ANALYSIS: The incremental cost-effectiveness of statin therapy compared with usual care in patients 75 to 84 years of age with previous myocardial infarction was $18800 per quality adjusted life-year (QALY). RESULTS OF SENSITIVITY ANALYSIS: On the basis of a probabilistic sensitivity analysis, there is a 75% chance that statin therapy costs less than $39800 per QALY compared with usual care. If the cost of statin therapy and efficacy of statin therapy at reducing myocardial infarction were set to their most favorable values, statin therapy cost $5400 per QALY; if cost and efficacy were set to their least favorable values, statin therapy cost $97800 per QALY. CONCLUSIONS: The cost-effectiveness ratios of statin therapy in older patients with previous myocardial infarction are reasonable under a wide variety of assumptions about drug efficacy, drug cost, and rates of cardiac and cerebrovascular events. Pending results of randomized, controlled trials of secondary prevention in patients in this age group, statin therapy seems to be as cost-effective as many routinely accepted medical interventions in this setting. PMID- 10819701 TI - Oral clonidine in postmenopausal patients with breast cancer experiencing tamoxifen-induced hot flashes: a University of Rochester Cancer Center Community Clinical Oncology Program study. AB - BACKGROUND: Hot flashes are the most frequently reported side effect of tamoxifen treatment. Although hormones are an effective treatment, their safety is questionable in women with breast cancer. It is therefore important to evaluate nonhormonal treatments for hot flashes. OBJECTIVE: To evaluate the effectiveness of oral clonidine for control of hot flashes associated with tamoxifen therapy in postmenopausal women with breast cancer. DESIGN: Randomized, double-blind, placebo-controlled clinical trial. SETTING: University of Rochester Cancer Center Community Clinical Oncology Program. PATIENTS: 194 postmenopausal women with breast cancer who were receiving adjuvant tamoxifen therapy. INTERVENTION: Oral clonidine hydrochloride, 0.1 mg/d, or placebo for 8 weeks. MEASUREMENTS: In a daily diary, patients recorded number, duration, and severity of hot flashes and overall quality-of-life score (on a 10-point scale) during a 1-week baseline period and during the 4th, 8th, and 12th weeks of the study. RESULTS: Patients in the placebo and treatment groups were similar in age, duration of tamoxifen use, reported frequency and duration of hot flashes at baseline, and dropout rates. One hundred forty-nine patients completed 12 weeks of follow-up. The mean decrease in hot flash frequency was greater in the clonidine group than in the placebo group after 4 weeks of treatment (37% compared with 20% [95% CI for difference, 7% to 27%]) and 8 weeks of treatment (38% compared with 24% [CI for difference, 3% to 27%]). Patients receiving clonidine were more likely than patients receiving placebo to report difficulty sleeping (41% compared with 21%; P = 0.02). A significant difference was seen in the mean change in quality-of life scores (0.3 points in the clonidine group compared with -0.2 points in the placebo group; P = 0.02) at 8 weeks, although the median difference was 0 in both groups. CONCLUSION: Oral clonidine, 0.1 mg/d, is effective against tamoxifen induced hot flashes in postmenopausal women with breast cancer. PMID- 10819702 TI - Antibody inhibitors to von Willebrand factor metalloproteinase and increased binding of von Willebrand factor to platelets in ticlopidine-associated thrombotic thrombocytopenic purpura. AB - BACKGROUND: Thrombotic thrombocytopenic purpura (TTP) affects 1 in 1600 to 1 in 5000 patients who receive ticlopidine, but little is known about the pathogenesis of this complication. OBJECTIVE: To investigate whether von Willebrand factor (vWF), which has been associated with idiopathic TTP, is involved in the pathogenesis of ticlopidine-associated TTP. DESIGN: Case series. SETTING: Three tertiary care, university-affiliated medical centers. PATIENTS: Seven patients who developed TTP 2 to 7 weeks after initiation of ticlopidine therapy. Controls were 7 consecutive patients without thrombocytopenia who had been receiving ticlopidine for 3 to 5 weeks and 10 randomly selected hospitalized patients. MEASUREMENTS: Platelet-bound vWF in patients' EDTA-anticoagulated whole blood samples; vWF proteinase activity in patients' plasma samples; inhibitory activity of IgG isolated from patients' plasma samples against the proteinase from the controls' plasma samples; and vWF multimeric patterns in patients' EDTA anticoagulated plasma samples. RESULTS: Binding of vWF to single platelets was increased in the three patients tested during the most thrombocytopenic phase of TTP episodes. Initial plasma samples from all seven patients lacked the largest vWF multimers and were severely deficient in vWF metalloproteinase. IgG molecules, isolated from plasma samples of five patients, inhibited metalloproteinase in plasma samples from the controls. In patients examined, these abnormalities resolved upon the remission that accompanied plasma exchange and discontinuation of ticlopidine therapy. CONCLUSION: In the patients who developed ticlopidine-associated TTP, autoantibodies to the vWF metalloproteinase were formed; this led to the same type of vWF abnormalities observed in patients with idiopathic acute TTP. The findings suggest that failure to process large and unusually large vWF multimers in vivo caused binding of vWF to platelets, systemic platelet thrombosis, and TTP. PMID- 10819703 TI - Absorption and hemodynamic effects of airway administration of adrenaline in patients with severe cardiac disease. AB - BACKGROUND: If intravenous access cannot be attained during resuscitation of adult patients, endotracheal application of at least 2 mg of adrenaline is recommended. However, the effects of this intervention have not yet been demonstrated in adults. OBJECTIVE: To demonstrate the effects of adrenaline administered through the airways. DESIGN: Prospective clinical trial. SETTING: Operating theater at university hospital. PATIENTS: 34 patients receiving implantable cardioverter defibrillators under general anesthesia. INTERVENTION: When mean arterial pressure decreased below 80 mm Hg, 100 times the effective central intravenous dose of adrenaline (mean +/- SD, 1.3+/-0.6 mg [range, 0.7 to 3 mg]) was administered over 5 seconds into the endotracheal tube or through a bronchial catheter. Ten forced ventilations followed. MEASUREMENTS: Hemodynamic variables were recorded with a polygraph recorder. Adrenaline levels were measured in 13 patients. RESULTS: Plasma levels and arterial pressure increased in all patients (P < 0.002). Higher plasma levels (P < 0.039) and greater arterial pressure (P < 0.001) were achieved with this method than with intravenous injection. The effects of adrenaline did not differ between the two airway routes. Sustained ventricular arrhythmia did not occur. CONCLUSION: These substantial effects support the standard recommendation to consider the airways as an alternate route for at least 2 mg of adrenaline during resuscitation. PMID- 10819704 TI - The first positive: computing positive predictive value at the extremes. AB - Computing the positive predictive value (PPV) of a wellknown test for a relatively common disease is a straight-forward exercise. However, in the case of a new test for a rare disorder; the extreme numbers involved-the very low prevalence of the disorder and the lack of previous false-positive results--make it difficult to compute the PPV. As new genetic tests become available in the next decade, more and more clinicians will have to answer questions about PPVs in cases with extreme prevalence, sensitivity, and specificity. This paper presents some tools for thinking about these calculations. First, a standard PPV calculation with rough estimates of the prevalence, sensitivity, and specificity is reviewed. The "zero numerator" problem posed by not having seen any false positive results is then discussed, and a Bayesian approach to this problem is described. The Bayesian approach requires specification of a prior distribution that describes the initial uncertainty about the false-positive rate. This prior distribution is updated as new evidence is obtained, and the updated expected false-positive rate is used to calculate PPVs. The Bayesian approach provides appropriate and defensible PPVs and can be used to estimate failure rates for other rare events as well. PMID- 10819705 TI - Accuracy of the Papanicolaou test in screening for and follow-up of cervical cytologic abnormalities: a systematic review. AB - PURPOSE: To evaluate the accuracy of conventional and new methods of Papanicolaou (Pap) testing when used to detect cervical cancer and its precursors. DATA SOURCES: Systematic search of English-language literature through October 1999 using MEDLINE, EMBASE, other computerized databases, and hand searching. STUDY SELECTION: All studies that compared Pap testing (conventional methods, computer screening or rescreening, or monolayer cytology) with a concurrent reference standard (histologic examination, colposcopy, or cytology). DATA EXTRACTION: Two reviewers independently reviewed selection criteria and completed 2 x 2 tables for each study. DATA SYNTHESIS: 94 studies of the conventional Pap test and three studies of monolayer cytology met inclusion criteria. No studies of computerized screening were included. Data were organized by cytologic and histologic thresholds used to define disease. For conventional Pap tests, estimates of sensitivity and specificity varied greatly in individual studies. Methodologic quality and frequency of histologic abnormalities also varied greatly between studies. In the 12 studies with the least biased estimates, sensitivity ranged from 30% to 87% and specificity ranged from 86% to 100%. CONCLUSIONS: Insufficient high-quality data exist to estimate test operating characteristics of new cytologic methods for cervical screening. Future studies of these technologies should apply adequate reference standards. Most studies of the conventional Pap test are severely biased: The best estimates suggest that it is only moderately accurate and does not achieve concurrently high sensitivity and specificity. Cost-effectiveness models of cervical cancer screening should use more conservative estimates of Pap test sensitivity. PMID- 10819706 TI - The Balanced Budget Act of 1997: its impact on U.S. teaching hospitals. AB - The Balanced Budget Act of 1997 had a profound impact on the financing and organization of many health care services. The Act disproportionately affected U.S. teaching hospitals, leading to substantial budget reductions in many institutions and the threat of cuts in major programs and services that teaching hospitals provide to communities. This paper examines the overall financial and organizational impact of the Balanced Budget Act on teaching hospitals and considers its effect on residency education. It also discusses to what degree the Balanced Budget Refinement Act of 1999 will mitigate these effects and posits other solutions to the serious financial issues facing teaching hospitals in the United States. PMID- 10819707 TI - In search of a good death: observations of patients, families, and providers. AB - Despite a recent increase in the attention given to improving end-of-life care, our understanding of what constitutes a good death is surprisingly lacking. The purpose of this study was to gather descriptions of the components of a good death from patients, families, and providers through focus group discussions and in-depth interviews. Seventy-five participants-including physicians, nurses, social workers, chaplains, hospice volunteers, patients, and recently bereaved family members-were recruited from a university medical center, a Veterans Affairs medical center, and a community hospice. Participants identified six major components of a good death: pain and symptom management, clear decision making, preparation for death, completion, contributing to others, and affirmation of the whole person. The six themes are process-oriented attributes of a good death, and each has biomedical, psychological, social, and spiritual components. Physicians' discussions of a good death differed greatly from those of other groups. Physicians offered the most biomedical perspective, and patients, families, and other health care professionals defined a broad range of attributes integral to the quality of dying. Although there is no "right" way to die, these six themes may be used as a framework for understanding what participants tend to value at the end of life. Biomedical care is critical, but it is only a point of departure toward total end-of-life care. For patients and families, psychosocial and spiritual issues are as important as physiologic concerns. PMID- 10819708 TI - Using cost-effectiveness analysis to target cholesterol reduction. PMID- 10819709 TI - On being a doctor. Shattered all. PMID- 10819710 TI - Antithrombotic therapy to prevent stroke in patients with atrial fibrillation. PMID- 10819711 TI - Chronic infection and coronary risk. PMID- 10819712 TI - Elevated ambulatory and normal office blood pressure. PMID- 10819713 TI - The sharer. PMID- 10819714 TI - Diagnosis of intercritical gout. PMID- 10819715 TI - Transmission of tuberculosis in a jail. PMID- 10819716 TI - Prophylactic fluconazole in liver transplant recipients. PMID- 10819717 TI - Role of communication skills training. PMID- 10819718 TI - Update in preventive medicine. PMID- 10819719 TI - Gated myocardial perfusion scan leading to diagnosis of unsuspected massive pulmonary embolism. PMID- 10819720 TI - Infection with hepatitis C virus genotype 4 is associated with a poor response to interferon-alpha. PMID- 10819721 TI - A common problem with therapy for HIV infection. PMID- 10819722 TI - Most gastric low-grade B-cell lymphomas of mucosa-associated lymphoid tissue persist after Helicobacter pylori eradication. PMID- 10819723 TI - Immediate cerebral angiography and mechanical fragmentation of cerebral embolus after percutaneous myocardial revascularization. PMID- 10819724 TI - Rapid bioprosthetic valve degeneration in a patient undergoing hemodialysis. PMID- 10819725 TI - From the ground up: a look at the role of foundations in health. PMID- 10819726 TI - [Staphylococcal toxic shock]. PMID- 10819727 TI - Microbiological efficacy of levofloxacin for treatment of community-acquired pneumonia due to Chlamydia pneumoniae. PMID- 10819728 TI - Papers presented at the 107th Scientific Session of the Western Surgical Association. Santa Fe, New Mexico, USA. November 15-17, 1999. PMID- 10819729 TI - Critical aortic stenosis in the neonate. Clinical assessment and surgical outcome in 18 patients. AB - OBJECTIVE: To assess neonates with aortic stenosis with early decompensation operated upon. (LCO) (CHF). METHODS: A and retrospective study analyzing 6 neonates with LCO, group I (GI), and 12 neonates with CHF, group II (GII). Clinical radiographic, electrocardiographic and echocardiographic findings also provided comparative bases for the study, as did surgical and evolutional findings. RESULTS: The mean ages at hospitalization and surgery (p = 0.0031) were 14.3 and 14.8 days in GI and 35.4 and 42.8 days in GII, respectively. Cardiac murmurs were more intense in GII (p = 0.0220). The aortic ring was smaller in GI (8.0 +/- 2.5mm) as compared to GII (11.4 +/ 1.4mm) (p = 0.2882). Ventricular function was reduced to 18 +/- 5.5% and 33.3 +/- 7.6% in GI and GII, respectively (p = 0.0162). Aortic atresia, however, was present only in 2 neonates in GI. Five of 6 patients in GI died but all patients in GII survived (p=0.0007). In the latter group, 84.6% of the patients were in functional class I (FC-I) in the long term follow-up, with moderate residual lesions in 6 neonates, discrete residual lesions in 4, and reoperation in 2. CONCLUSION: Aortic stenosis is a severe anomaly of the neonate, whose immediate evolution depends on the pre-operative anatomic and functional findings, and the late evolution essentially depends on the anatomic features of the valve. PMID- 10819730 TI - Leisure-time physical activity and structural arterial properties of the common carotid artery. PMID- 10819731 TI - Scholarship for a new era. PMID- 10819732 TI - Meeting the challenges of the 21st century. PMID- 10819733 TI - Old wine in new bottles: delivering nursing in the 21st century. PMID- 10819734 TI - Integrated literature review of continuity of care: Part 1, Conceptual issues. AB - PURPOSE: Continuity of patient care is a fundamental tenet of professional nursing, yet comprehension of the concept and related issues remains elusive. The purpose of this study was to explore issues associated with definitions, related concepts, and research methods of continuity of care through systematic literature-based study. In Part 1 of this two-part series, definitions and related concepts, factors, and variables associated with continuity of care were explored. DESIGN: Qualitative, integrated literature review of a sample of 38 nursing research articles about continuity of care, 1990-1995. METHODS: Ganong's (1987) stages of an integrative research review guided this study. A data collection tool was developed and pilot tested, and rules for data analysis and interpretation were established. Research articles were systematically analyzed and reported using descriptive analysis. FINDINGS AND CONCLUSIONS: No consensus was found in the literature about the conceptual definition of continuity of care. Continuity of patient care is a multifactorial concept affected by environmental influences, communication, patient, professional, and system factors. PMID- 10819736 TI - Sleep in acute care settings: an integrative review. AB - PURPOSE: To report an integrative review about sleep patterns, factors that influence sleep, and sleep interventions in acutely ill hospitalized adults; discuss methodological challenges associated with studying sleep in this population; and propose future research. ORGANIZING FRAMEWORK: Research literature on sleep patterns of acutely ill hospitalized adults, obtained using computerized searches of Medline and CINAHL, was reviewed. Studies of patients on intensive care and medical-surgical units conducted in a range of health disciplines over a span of almost 30 years were included. Studies were organized into the following categories: descriptive studies of sleep using objective and subjective measures; correlational studies of personal, health status, and environmental variables and sleep; and intervention studies. FINDINGS: Sleep disturbance is a common but highly variable experience during acute care hospitalization. Multiple personal, health status, and environmental factors are related to sleep disturbance. Behavioral interventions show promise as a means to modify factors related to sleep. Few studies have shown the outcomes of sleep disturbance in acute care settings. CONCLUSIONS: More systematic research is needed to determine correlates of sleep disturbance in acutely ill hospitalized adults. Such studies can help investigators to identify patients who are most at risk for sleep disturbance and to provide the theoretical and conceptual bases for sleep-promoting interventions. Methodological challenges include characteristics of acute care patients and hospital environments, as well as the complexity of measuring sleep. PMID- 10819735 TI - Information needs of elderly postsurgical cancer patients during the transition from hospital to home. AB - PURPOSE: To describe information needs of elderly postsurgical cancer patients. Few reports have described learning needs of postsurgical cancer patients during the initial weeks after hospitalization. Responding to information needs during the transition to home is important for helping patients and families to appropriately manage illness. DESIGN: Descriptive, using records kept for patients assigned to the experimental group in a larger study completed in 1996. The sample was 148 American patients surgically treated for a new diagnosis of prostate, breast, gastrointestinal, lung, or head and neck cancer. Information needs were identified from topics addressed by advanced practice nurses during 4 week episodes of home care. METHODS: Content analysis of 3,280 statements of teaching interventions. FINDINGS: Teaching themes were instructing on postoperative self-care, advising on symptom management, clarifying the illness experience, discussing psychological responses, and preparing patients and families to coordinate follow-up care. Topics included describing the expected course of illness, informing about community resources, identifying events that require physician notification, and advising on pain management. Topic emphasis varied by site of cancer. CONCLUSIONS: Information needs of elderly postsurgical cancer patients during transition from hospital to home are extensive. Teaching interventions ranged from giving concrete instructions about care of a surgical wound to interpreting complex information about options for cancer treatment. Elderly postsurgical cancer patients need informational support during the transition from hospital to home. PMID- 10819737 TI - Being strong: how black West-Indian Canadian women manage depression and its stigma. AB - PURPOSE: To discover how women from a nondominant cultural background (West Indian) experience and manage depression. DESIGN: Explanatory using grounded theory. METHODS: Semistructured interviews were conducted with 12 Black West Indian Canadian women who experienced depression. Between 1994 and 1996, the first author engaged in participant observation. FINDINGS: The women used the basic social process they called "being strong" to manage or ameliorate depression. Being strong included "dwelling on it," "diverting myself," and "regaining my composure." For most of the women, the range of available life choices was limited to the three processes; however, a few engaged in "trying new approaches." These women were less limited in their range of cultural and behavioral boundaries than were the others, and began tentatively to explore other options for themselves. CONCLUSIONS: Black West-Indian Canadian women in this study managed their depression in culturally defined ways by being strong and not showing vulnerability. Because being strong was also evident in a previous study of dominant-culture women as a prelude to depression, the process may be widespread in women prone to depression. The findings provide helpful information for intervening in an unfamiliar culture. PMID- 10819738 TI - An historical analysis of disclosure of terminal status. AB - PURPOSE: To examine the concept of disclosure of terminal status and trace its development and implementation over time. An individual's ability to make appropriate end-of-life decisions, exert some control over the place and manner of death, and prepare self and significant others for this loss depends on knowing that life is drawing to a close. ORGANIZING FRAMEWORK: An analysis of disclosure of terminal status was performed within the context of American social, political, and cultural history from 1930 to 1990. METHODS: A combination of manual and computerized search mechanisms was performed utilizing popular, nursing, medical, sociological, psychological, and thanatological literature from 1930 to 1990. The articles and books obtained through these searches were scanned and those articles and books that dealt with health care disclosure issues were selected. Works included in the review were 149 articles and 22 books. FINDINGS: A long-standing tradition of nondisclosure exists in the medical profession. Societal events in the 1950s and 1960s influenced individuals' perceptions of death, individual rights, and physician responsibility, and culminated in some change in this tradition in the 1970s. The magnitude of this change is unknown. Current practice has not been investigated. This topic is rarely addressed in nursing literature even though nursing care is influenced by physicians' practice. CONCLUSIONS: A tradition of nondisclosure prohibits patients from managing the end of their lives and making choices about their manner of death. Nursing care of terminally ill patients is compromised when patients are uninformed. PMID- 10819739 TI - Developing a database to describe the practice patterns of adult nurse practitioner students. AB - PURPOSE: To describe the practice patterns of adult nurse practitioner students using a database composed of core health data elements and standardized nursing language. DESIGN: Descriptive study of 3,733 patient visits documented by 19 adult nurse practitioner students in the academic year 1996-1997. METHODS: A database was designed for documenting the full scope of practice of adult nurse practitioner students by use of core health data elements and the standardized nursing languages of NANDA and NIC. Nurse practitioner students used the database to document every clinical encounter during their final clinical year of study. Most visits occurred in ambulatory care settings in a midwestern American city. FINDINGS: Based on the American Medical Association's Evaluation/Management coding system, data indicated that 50% of visits were classified as problem focused, while 31.9% were expanded, 10% were detailed, and 8.1% were comprehensive. The most frequently occurring NANDA diagnoses were pain, health seeking behavior, altered health maintenance, and knowledge deficit. The most frequently reported nursing intervention classifications (NIC) were patient education, drug management, information management, and risk management. CONCLUSIONS: Using standardized nursing language to describe clinical encounters made visible the complex clinical decision-making patterns of adult nurse practitioner students. Systematic use of a database designed for documenting the full scope of practice of nurse practitioner students showed the applicability of standardized nursing language to advanced practice nursing contexts. PMID- 10819740 TI - Beyond the significance test in administrative research and policy decisions. AB - PURPOSE: To describe confidence interval (CI) analysis and show how it can be used in administrative decisions. ORGANIZING CONSTRUCT: Statistical significance testing should be supplemented, if not replaced, by effect size (ES) estimation and confidence interval analysis. Hypothesis testing based on the statistical significance test is the dominant paradigm in statistics; however, this approach has inherent problems which can ultimately diminish the usefulness of research for operational decisions. After identifying major difficulties with significance testing, the authors use hypothetical examples to demonstrate how ES and CI analysis provide more informative answers to nursing administrative research questions. CONCLUSIONS: CI analysis provides the basis for both improving the interpretation of findings from individual studies and for facilitating the analysis of cumulative evidence. By clarifying the meaning of results, CI analysis can increase the relevance and usefulness of research for health care executives and practitioners. PMID- 10819741 TI - Comparison of nurse practitioner and family physician relative work values. AB - PURPOSE: With the enactment of the Balanced Budget Act of 1997, American nurse practitioners were granted direct Medicare reimbursement for Part B services. Payment structures in fee-for-service and managed care systems are physician based, leading to difficulties in constructing payments for other health care professionals. The purpose of this pilot study was to examine the feasibility of using nurse practitioner data for specifying relative work values in the Medicare Fee Schedule for three office-visit codes. DESIGN: An exploratory survey was designed to establish relative work values using magnitude-estimation scaling. Nurse practitioners (N = 43) responded to a structured questionnaire in a national mail survey. Physician data (N = 46) were obtained from a computerized database from the American Academy of Family Physicians. METHODS: The methods used in this study were the same as the process used by the American Medical Association and the Health Care Financing Administration to establish relative work values in the Medicare Fee Schedule. Respondents established relative work values for three Current Procedural Terminology (CPT) codes for office visits (99203, 99213, 99215) commonly billed in primary care practice. Each CPT code descriptor and associated vignette were compared with reference services germane to the practice of nurse practitioners and family physicians, using magnitude estimation scaling. To establish relative work values for each code, respondents were asked to consider the time to provide the service and intensity of the work involved for each CPT code. FINDINGS: No significant differences between nurse practitioners and family physicians were found in the three CPT codes for relative work values and intensity. Nurse practitioners estimated significantly (p < .01) higher intraservice (face to face) time with patients than did family physicians, and family physicians estimated significantly (p < .05) higher pre service time for two codes and significantly (p < .05) higher postservice times for three codes. CONCLUSIONS: Nurse practitioner relative work values did not differ significantly from family physician relative work values. Although the sample sizes were small, the significance of the findings support the need for further research with large data sets and additional CPT codes. Such studies could then be used as a basis for decisions about Medicare payment and public policy. PMID- 10819742 TI - Research priorities for staffing, case mix, and quality of care in U.S. nursing homes. PMID- 10819743 TI - Avoiding gender and minority barriers to NIH funding. AB - PURPOSE: To inform investigators of the National Institutes of Health (NIH) guidelines for the inclusion of women and minorities as subjects in clinical research and provide tips on avoiding barriers to federal funding. METHODS: Information about gender and minority barriers to funding was obtained by reviewing NIH policies, information submitted by applicants, and comments provided by NIH reviewers. ORGANIZING FRAMEWORK: A brief history of the goals of U.S. federal legislation to increase opportunities for obtaining information to help enhance health and disease treatment for all Americans and to detect and account for ethnic and gender differences is described. Examples were derived from NIH Summary Statement comments of initial review groups. CONCLUSIONS: Applications submitted to NIH are being barred from initial funding, even with excellent priority scores, if the plan for inclusion of women and minorities is judged to be scientifically unacceptable by the initial review group. This situation is correctable and avoidable. PMID- 10819744 TI - Nursing publications outside the United States. AB - PURPOSE: To replicate a 1992-1993 study of the characteristics of English language nursing journals originating in countries other than the United States and to compare findings. Such information heightens awareness of publishing opportunities globally and enhances dissemination of information throughout the world. DESIGN: Descriptive survey with a questionnaire mailed to 159 editors of nursing and nursing-related journals. Data about the publication year, 1995, were collected from April 1996 through March 1997. METHOD: A 38-item questionnaire pertaining to journal, readership, manuscript review, and journal staff characteristics was used. FINDINGS: Information about 82 journals from 13 countries was collected with an overall response rate of 55%. In the 1992-1993 study the United Kingdom, Canada, and Australia accounted for the largest percent of publications. With few exceptions, results of the 1996-1997 and the earlier survey are remarkably similar. Differences include a higher total circulation, changes in circulation among journal categories, and more publications offering services to authors. Two main reasons for manuscript rejection continue to be that a manuscript is poorly written or poorly developed. CONCLUSIONS: Increased awareness of non-U.S. publishing outlets can lead to the acceptance of informative and well-written manuscripts and ultimately to the dissemination of information and knowledge. PMID- 10819745 TI - Trends in nursing education in China. AB - PURPOSE: To explore the current trends and development of nursing education in China. Nursing education in China has developed rapidly in alignment with economic growth of this vast country. DESIGN: Descriptive, using 345 manuscript abstracts from various regions of China, submitted for the 1996 Sino-Hong Kong Educational Conference. METHODS: Content analysis of manuscript abstracts was done to describe the trends. Codes were determined after several rounds of independent analysis. FINDINGS: Development of nursing education in China appears in line with global trends. The preparation of a pragmatic nurse was emphasized by nurse educators who also advocate total patient care and the integration of traditional Chinese medicine in nursing education. Areas seldom mentioned, however, were student-centered approaches to learning, advanced practice, and the evaluation of educational processes and outcomes. CONCLUSIONS: Nurse educators in mainland China appear aware of the dynamics and challenges in health care, and the need for preparing a new generation of nurses to face these challenges. Educators in China need to adopt new approaches in teaching and learning to prepare nurses to function in a wide range of health care settings, and to monitor and evaluate educational outcomes. PMID- 10819746 TI - Gene expression in abnormal ovarian structures of ewes homozygous for the inverdale prolificacy gene. AB - Animals heterozygous (I+) for the Inverdale prolificacy gene (FecX(I)) have an increased ovulation rate whereas those homozygous (II) for FecX(I) are infertile with "streak" ovaries and follicular development arrested at the primary (type 2 follicle) stage. The streak ovaries also contain small oocyte-free nodules with granulosa-like cells and often tumor-like structures. It has been hypothesized that these abnormal structures are of granulosa cell origin, and the aim of this study was to determine whether genes normally expressed in granulosa cells are also expressed in the nodules and tumor-like structures. The mRNAs encoding c-kit and its ligand stem cell factor (SCF), FSH receptor (FSH-R), follistatin, alpha inhibin subunit, and the beta(A)- and beta(B)-activin/inhibin subunits were localized in ovaries of ewes with 0 (++), 1 (I+), or 2 (II) copies of the FecX(I) gene (n = 4-9 animals per genotype per gene) using in situ hybridization. Ontogeny of expression of all mRNAs examined was similar between ++ and I+ ewes. Expression of c-kit mRNA was observed in the oocyte of all follicular types present in ++, I+, and II ewes. Moreover, granulosa cells of type 2 (II) and type 2 and larger follicles (++, I+) expressed SCF mRNA. The mRNAs encoding FSH-R, follistatin, alpha-inhibin subunit, and beta(B)-activin/inhibin subunit were identified in type 3 and larger follicles of ++ and I+ ewes but not in follicles of II ewes that were only at the type 1, 1a, or 2 stages of development. However, the cells within the oocyte-free nodules of II ewes expressed all of these genes. The mRNAs encoding c-kit and beta(A)-activin/inhibin subunit were not observed in granulosa cells until antrum formation (type 5 follicles) or in the nodules of II ewes. Tumors from 4 ewes were obtained and classified as cystic, semisolid, or solid structures containing granulosa-like cells or as solid structures containing predominately fibroblast- and luteal-like cells. Often, two tumors were present on the same ovary. Tumors containing granulosa-like cells (n = 3-4 per gene) expressed the mRNAs encoding alpha-inhibin subunit, beta(A)-, and beta(B)-activin/inhibin subunits, follistatin, and the FSH-R but did not contain detectable amounts of mRNA for c-kit or SCF. Tumors composed predominately of fibroblast- and luteal-like cells expressed very low levels of SCF mRNA; of the other mRNAs examined, none were detected. Also, none of the genes examined were found to be expressed by the surface epithelium, theca externa, fibroblast, or vascular cells within the ovary of animals of any genotype. These findings are consistent with the hypothesis that the somatic cells in oocyte-free nodules and tumor-like tissue in II ewes originate from the granulosa cells of the small follicles. PMID- 10819747 TI - Expression of growth and differentiation factor-9 in the ovaries of fetal sheep homozygous or heterozygous for the inverdale prolificacy gene (FecX(I)). AB - Abnormal follicular and oocyte growth in ovaries of sheep homozygous (II) for the Inverdale gene, FecX(I), suggest that this gene may influence a fundamental event in initiation of folliculogenesis, with two copies of the gene inhibiting growth at the primordial/primary stage. In addition, striking similarities in ovarian morphology between mice deficient in growth and differentiation factor-9 (GDF-9) and II sheep suggest a relationship between the FecX(I) gene and GDF-9 function in the ovary. Therefore, it was hypothesized that GDF-9 mRNA expression would be inhibited in ovaries of II fetal sheep. To test this hypothesis, in situ hybridization was used to characterize GDF-9 mRNA expression in ovaries of homozygous (II), heterozygous (I+), and control (++) fetal sheep at Day 135 of gestation. GDF-9 mRNA expression was localized exclusively to oocytes from the type 1 follicle stage onward in all genotypes and is the first demonstration of GDF-9 mRNA expression in ovaries of fetal sheep. In addition, GDF-9 mRNA expression was detected in oocytes of abnormal type 2 follicles in the ovaries of II sheep. Thus, it does not appear that inhibition of GDF-9 gene expression is the mechanism of action whereby the FecX(I) gene exerts its influence. However, the possibility of translation at specific stages of follicular development cannot presently be ruled out. In addition, the FecX(I) gene may be involved, either directly or indirectly, in regulating expression of receptors for GDF-9. At present, however, neither the FecX(I) gene product nor the GDF-9 receptor has been isolated or characterized. PMID- 10819748 TI - Involvement of p38 mitogen-activated protein kinase activation in bromocriptine induced apoptosis in rat pituitary GH3 cells. AB - Bromocriptine, a dopamine D(2) receptor agonist, is a therapeutic agent for patients with prolactinoma and hyperprolactinemia. In this study we demonstrated that bromocriptine induced activation of p38 mitogen-activated protein (MAP) kinase, with concomitant induction of apoptosis in rat pituitary adenoma cell line GH3 cells. Treatment of GH3 cells for 48 h with bromocriptine increased the p38 MAP kinase activity up to 3- to 5-fold and simultaneously increased the number of apoptotic cells. Inclusion in the medium of SB212090 or SB203580, specific p38 MAP kinase inhibitors, completely abolished the bromocriptine induced activation of p38 MAP kinase and significantly reduced the number of apoptotic cells. The bromocriptine-induced p38 MAP kinase activation was not prevented by S(-)-eticropride hydrochloride, a specific D(2) receptor antagonist. Treatment with either epidermal growth factor (EGF) or thyrotropin-releasing hormone (TRH), which stimulates p44/42 MAP kinase, rescued cells from the bromocriptine-induced apoptosis, with concomitant inhibition of the bromocriptine induced p38 MAP kinase activation. These results suggest that bromocriptine induces apoptosis in association with p38 MAP kinase activation, and that the p44/42 MAP kinase signaling through EGF and TRH receptors has an opposing effect on p38 MAP kinase activation as well as on apoptosis induced with bromocriptine in GH3 cells. PMID- 10819749 TI - Blood chemistry, nutrient metabolism, and organ weights in fetal and newborn calves derived from in vitro-produced bovine embryos. AB - Calves born after in vitro production (IVP) of embryos often show reduced perinatal viability. The present experiment investigated a series of physiological variables in the immediate prenatal and postnatal period of IVP dairy calves. Fetal IVP and control calves (each n = 7) were prepared with vascular catheters at 248+/-1 day gestation (term = 280 days), and blood samples were taken for five days before premature delivery by cesarean section. IVP fetuses compared with controls had significantly elevated arterial hemoglobin and oxygen content (8.41 vs. 7.52% and 5.75 vs. 3.79%, respectively) whereas lactate level was lowered (1.89 vs. 2.26 mM). The umbilical venous-arterial concentration differences in oxygen, lactate, and glucose indicated that IVP fetuses relied more on lactate and less on glucose as oxidative substrates. The fetal glucose tolerance, and the basal and adrenocorticotropin-stimulated cortisol levels were similar between the groups. In the immediate postnatal period, IVP calves showed elevated venous blood pH (7.294 vs. 7.270), hemoglobin (9.06 vs. 8.25%), oxygen contents (6.33 vs. 4. 64%), K(+) levels (4.89 vs. 4.56 mM), and rectal temperature (38.9 vs. 37.4 degrees C), and lowered blood Na(+) (139.9 vs. 141.0 mM), Cl(-) (100.2 vs. 103.1 mM) and glucose levels (2.86 vs. 3.11 mM). There were no differences in body dimensions and organ weights, except that the fore legs and hind legs were slightly longer in the IVP group (76.1 vs. 72.4 cm and 93.4 vs. 88.8 cm, respectively). Although prenatal and neonatal IVP calves differed from control calves in a number of variables, the effects were relatively minor and provide no direct evidence for the hypothesis that IVP calves have an impaired capacity to adapt to life ex utero. In fact, several parameters indicated enhanced rather than retarded maturation of IVP calves when data from premature calves were compared with data from a group of control calves delivered at term. PMID- 10819750 TI - Passive immunization with anti-laminin immunoglobulin G modifies the integrity of the seminiferous epithelium and induces arrest of spermatogenesis in the guinea pig. AB - In the testis, the base of the Sertoli cells is in contact with the basement membrane matrix, in which the laminins constitute the major noncollagenous components. We have previously demonstrated that antibodies against a preparation enriched in basement membranes of seminiferous tubules (STBM) or a noncollagenous fraction of STBM passively transferred induced modifications to the basement membranes and focal sloughing of the seminiferous epithelium in the rat. In the present report, we tested the effect of passive immunization with anti-laminin IgG on the limiting membrane of the seminiferous tubules, spermatogenesis, and maintenance of the blood-testis barrier in the adult guinea pig. Rabbit antibodies to laminin 1 (IgG fraction) were injected in adult male guinea pigs (GP). Nonimmunized GP and GP immunized with normal rabbit serum IgG were used as controls. Measurements of variations in the diameter and lumen of the tubules and in the size of individual components of the tubular limiting membrane showed that the highest percentage of tubules with reduced lumen occurred 30 days after passive immunization with anti-laminin, when the limiting membrane was thickest and lesions to the seminiferous epithelium were most severe. The lesions included thickening of the limiting membrane, infolding in the basal lamina, deposits of immune complexes coincident with sloughing of pachytene spermatocytes and spermatids, and vacuolization of the Sertoli cells. Mononuclear cell infiltration of the tubules was rare. Permeability tracer studies revealed that Sertoli cell tight junctions remained impermeable. Fifty and 80 days after treatment, the basement membrane of the tubules and the progression of the spermatogenesis were normal. Passive immunization with anti-laminin IgG provided a valuable experimental model for the in vivo study of the influence of the basement membrane on the issue of spermatogenesis and the integrity of the seminiferous epithelium. PMID- 10819751 TI - Characterization of temporal and cell-specific changes in transcripts for prostaglandin E(2) receptors in pseudopregnant rat endometrium. AB - In the rodent uterus, prostaglandin E(2) (PGE(2)) is believed to have a major role in implantation and decidualization. The present study investigated the temporal and hormonal control of mRNA expression for the four E-prostanoid (EP(1 4)) receptors in the rat endometrium. For Northern blot analysis and in situ hybridization, samples were obtained from rats on Days 1-10 of pseudopregnancy or from rats differentially sensitized for the decidual cell reaction with estradiol. No EP(1) mRNA signal was detected. Endometrial EP(2) and EP(3) mRNA levels increased to a maximum on Day 5, and the mRNAs were localized to the luminal epithelium at the antimesometrial pole, and in the endometrial stroma and glandular epithelium, respectively. Endometrial EP(4) mRNA levels were unchanged on Days 1-5, but the mRNA was concentrated in the antimesometrial endometrial stroma on Day 5. Cell-specific expression of EP(2), EP(3), and EP(4) on Day 5 was dependent upon a dose of estradiol given on Day 4 that induced differential uterine sensitization on Day 5. After the application of a deciduogenic stimulus on Day 5, mRNA levels for these receptors decreased significantly, while in nonstimulated horns they remained elevated. Overall, these results support a role for PGE(2) in the onset of receptivity and initiation of decidualization in the rat. PMID- 10819752 TI - Differential effects of culture on imprinted H19 expression in the preimplantation mouse embryo. AB - The H19 gene is imprinted with preferential expression from the maternal allele. The putative imprinting control region for this locus is hypermethylated on the repressed paternal allele. Although maternal-specific expression of H19 is observed in mouse blastocysts that develop in vivo, biallelic expression has been documented in embryos and embryonic stem cells experimentally manipulated by in vitro culture conditions. In this study the effect of culture on imprinted H19 expression and methylation was determined. After culture of 2-cell embryos to the blastocyst stage in Whitten's medium, the normally silent paternal H19 allele was aberrantly expressed, whereas little paternal expression was observed following culture in KSOM containing amino acids (KSOM+AA). Analysis of the methylation status of a CpG dinucleotide located in the upstream imprinting control region revealed a loss in methylation in embryos cultured in Whitten's medium but not in embryos cultured in KSOM+AA. Thus, H19 expression and methylation were adversely affected by culture in Whitten's medium, while the response of H19 to culture in KSOM+AA approximated more closely the in vivo situation. It is unlikely that biallelic expression of H19 following culture in Whitten's medium is a generalized effect of lower methylation levels, since the amount of DNA methyltransferase activity and the spatial distribution of Dnmt1 protein were similar in in vivo-derived and cultured embryos. Moreover, imprinted expression of Snrpn was maintained following culture in either medium, indicating that not all imprinted genes are under the same stringent imprinting controls. The finding that culture conditions can dramatically, but selectively, affect the expression of imprinted genes provides a model system for further study of the linkage between DNA methylation and gene expression. PMID- 10819753 TI - A basic 18-amino acid peptide contains the polysulfate-binding domain responsible for activation of the boar proacrosin/acrosin system. AB - Proacrosin is the zymogen of acrosin, a serine protease localized in the acrosomal matrix of mammalian sperm. Proacrosin/acrosin binds to solubilized zona pellucida glycoproteins (ZPGs) and various polysulfates in a non-enzymatic mechanism. In addition, both polysulfates and ZPGs induce proacrosin activation once they bind to the polysulfate-binding domain (PSBD) of the enzyme. We show here that the peptide (43)IFMYHNNRRYHTCGGILL(60) inhibited the proacrosin activation induced by either fucoidan or ZPGs. In addition, the peptide was recognized by the monoclonal antibody C5F10, which is directed against the PSBD region. Our data suggest that the PSBD is composed of many "subsites" that may or may not interact with each other. PMID- 10819754 TI - Posttranscriptional regulation of human leukocyte antigen G during human extravillous cytotrophoblast differentiation. AB - Human maternal tolerance to a semiallogenic fetus may be maintained, in part, by the unusual expression pattern of antigen-presenting molecules in placental trophoblast cells. Extravillous cytotrophoblast (EVC) cells, which invade the maternal decidua, express high levels of human leukocyte antigen G (HLA-G), a nonclassical, major histocompatibility complex (MHC) class I molecule. HLA-G transcripts have been detected in tumors and other tissues, yet protein accumulation is rare. We show that, within EVC cells themselves, the mRNA is more broadly expressed than the protein. Specifically, accumulation of HLA-G protein was markedly delayed during EVC cell differentiation. To elucidate this mechanism, we performed a comprehensive analysis comparing the expression of HLA G and proteins essential for MHC class I expression at the cell surface. The transporter for antigen processing proteins TAP1 and TAP2, as well as tapasin and beta(2)-microglobulin, appeared to be coordinately expressed throughout EVC cell columns. Strikingly, they all accumulated well in advance of the HLA-G protein but concurrently with its mRNA. A similar delay in the accumulation of the HLA-G protein was observed in vitro, using cultures of chorionic villi. We conclude that posttranscriptional regulation of HLA-G is fundamental to EVC cell development and is achieved independently of the peptide loading system. This represents a novel mechanism of MHC class I regulation. PMID- 10819755 TI - Evidence that luteinizing hormone-releasing hormone statin from ovine rete testis fluid is immunologically related to alphaC inhibin. AB - LHRH Statin is a putative gonadal protein that increases the interval between two consecutive LHRH pulses. The present work was aimed at analyzing the immunological homology between LHRH Statin and the N-terminal region of the alphaC subunit of inhibin. Thus, rete testis fluid (RTF) proteins were purified by immunoaffinity chromatography using antibodies against residues 1-7 plus 7-30 (experiment 1, A-fractions) and 14-28 of the alphaC inhibin subunit (experiment 2, B-fractions), and the LHRH Statin activity of the fractions was examined by intracerebroventricular administration in castrated rams followed by RIA of plasma LH levels in 15-min blood samples. Fractions that bound to the immunoaffinity column with low affinity were eluted with 0.5 M NaCl, pH 7.4 ( F2); then highly bound fractions were eluted sequentially in acidic (pH 2.5, -F3) followed by basic conditions (pH 11.5, -F4). In experiment 1, RTF (40 microg, n = 4) and highly bound fractions (A-F3, 30 ng, n = 8, 150 ng, n = 3; A-F4, 120 ng, n = 5) decreased LH mean plasma levels between 4 and 6 h after injection by 39%, 29%, 43%, and 37%, respectively (P<0.001 to 0.01), while the weakly bound fractions (A-F2, 180 ng, n = 4) and albumin control (40 microg, n = 4) had no activity. In experiment 2, RTF (100 microg, n = 4) and B-F3 (100 ng, n = 3) decreased plasma LH levels by 48% and 38%, respectively (P<0.001 to 0.05), whereas B-F4 (100 ng, n = 4) and albumin control (100 microg, n = 4) had no effect. A fraction obtained from B-F3 by gel filtration had significant LHRH Statin activity (63%, n = 6, P<0.001). PAGE with colloidal gold staining revealed 3 high molecular weight bands and 5 low molecular weight bands in B-F3. The 3 high molecular weight bands were shown to belong to the clusterin family and did not appear to have LHRH Statin activity. The 5 low molecular weight bands were all labeled by anti-alphaC inhibin antibodies. Collectively, these results strongly suggest that LHRH Statin has some homology with the 14-28 alphaC inhibin sequence. PMID- 10819756 TI - Differential localization of conventional protein kinase C isoforms during mouse oocyte development. AB - Protein kinase C (PKC), the major cell target for tumor-promoting phorbol esters, plays a central role in signal transduction pathways. In many biological systems where Ca(2+) serves as a second messenger, regulatory control is mediated by PKC. The activation of PKC depends on its binding to RACK1 receptor, which is an intracellular protein anchor for activated PKC. We demonstrate that the conventional PKC (cPKC) isoforms, PKC-alpha, PKC-betaI, and PKC-betaII, as well as RACK1, are expressed in mouse oocytes (germinal vesicle [GV]) and mature eggs (metaphase II [MII]). In GV oocytes, PKC-alpha, PKC-betaII, and RACK1 were uniformly distributed in the cytoplasm, while PKC-betaI was localized in the cytoplasm and in the plasma membrane as well. Treatment of GV oocytes with the biologically active phorbol ester, 12-o-tetradecanoyl phorbol-13-acetate (TPA), resulted in a rapid translocation of the cytosolic PKC-alpha, but not PKC-betaI, PKC-betaII, or RACK1, to the plasma membrane. This was associated with inhibition of GV breakdown. In MII eggs (17 h post-hCG), PKC-alpha was uniformly distributed in the cytoplasm while PKC-betaI and -betaII were distributed in the cytoplasm and in the plasma membrane as well. Treatment with TPA resulted in a rapid translocation of PKC-alpha from the cytoplasm to the plasma membrane and a significant decrease of PKC-betaI throughout the cytoplasm, while it also remained in the cell periphery. No change in the distribution of PKC-betaII or RACK1 was observed. TPA also induced pronucleus formation. Physiological activation of MII eggs by sperm induced cortical granule exocytosis associated with significant translocation of PKC-alpha and -betaI, but not -betaII, to the plasma membrane. Overall, these results suggest a possible involvement of cPKC isoforms in the mechanism of mouse oocyte maturation and egg activation. PMID- 10819757 TI - Prolactin-induced expression of intercellular adhesion molecule-1 and the accumulation of monocytes/macrophages during regression of the rat corpus luteum. AB - Intercellular adhesion molecule-1 (ICAM-1) is thought to facilitate the recruitment and migration of monocytes/macrophages to sites of inflammation. Here we investigated whether the luteolytic effect of prolactin in the hypophysectomized rat is associated with the expression of ICAM-1. In addition, we examined the effect of exogenous testosterone (or its potential conversion to estradiol endogenously) on the corpus luteum to address recent speculation that ovarian steroids might augment luteal regression. Immature, 30-day-old rats were ovulated with eCG and hCG and then hypophysectomized; this resulted in a single cohort of persistent corpora lutea. The rats were assigned randomly into four treatment groups: vehicle treatment without or with testosterone (VEH-T4, VEH+T4) and prolactin treatment without or with testosterone (PRL-T4, PRL+T4). Corpora lutea of control rats exhibited minimal ICAM-1 staining and contained relatively few monocytes/macrophages. In contrast, corpora lutea of prolactin-treated rats exhibited prominent ICAM-1 staining and contained numerous monocytes/macrophages. Testosterone did not overtly affect ICAM-1 staining, numbers of monocytes/macrophages, or concentrations of plasma progestins (progesterone and 20alpha-dihydroprogesterone) in either VEH or prolactin treatment groups; notwithstanding, luteal weights increased significantly in response to testosterone in VEH+T4 rats compared to VEH-T4 rats and prolactin-treated rats. We conclude that ICAM-1 expression and monocyte/macrophage accumulation are associated with prolactin-induced luteal regression in the rat and that these aspects are not influenced by testosterone. PMID- 10819758 TI - Production of male cloned mice from fresh, cultured, and cryopreserved immature Sertoli cells. AB - Although it is generally accepted that relatively high efficiencies of somatic cell cloning in mammals can be achieved by using donor cells from the female reproductive system (e.g., cumulus/granulosa, oviduct, and mammary gland cells), there is little information on the possibility of using male-specific somatic cells as donor cells. In this study we injected the nucleus of immature mouse Sertoli cells isolated from the testes of newborn (Days 3-10) males into enucleated mature oocytes in order to examine the ability of their nuclei to support embryonic development. After activation of the oocytes that had received the freshly recovered immature Sertoli cells, some developed into the morula/blastocyst stage, depending on the age of the donor cells (22.0-37.4%). When transferred into pseudopregnant females, 7 (3.3%, 7 of 215) developed into normal pups at term. Nuclear transfer of immature Sertoli cells after 1 wk in culture also produced normal pups after embryo transfer (3.1%, 2 of 65). Even after cryopreservation in a conventional cryoprotectant solution, their ability as donor cells was maintained, as demonstrated by the birth of cloned young (6.7%, 7 of 105). Immature Sertoli cells transfected with green fluorescent protein gene also supported embryo development into morulae/blastocysts, which showed specific fluorescence. This study demonstrates that immature Sertoli cells, male-specific somatic cells, are potential donors for somatic cell cloning. PMID- 10819759 TI - Activin, inhibin, and follistatin in zebrafish ovary: expression and role in oocyte maturation. AB - Activins, inhibins, and follistatins are important regulators of mammalian reproduction. However, their roles in lower vertebrates are poorly understood. In this study, we examined the expression of activin A, inhibin A, and follistatins in the zebrafish ovary and determined their role in final oocyte maturation. Using reverse transcription-polymerase chain reaction with primers specific for activin/inhibin beta(A) subunit and for follistatins, we detected DNA fragments of the expected size, which, upon sequencing, conformed to activin/inhibin beta(A) and follistatin. Western blot analysis using an antibody against activin/inhibin beta(A) subunit revealed two bands with sizes similar to those of activin A and inhibin A. The expression of follistatins was also confirmed by Western blot analysis. These results suggest that activin A, an inhibin A-like molecule, and follistatins are expressed in the zebrafish ovary. In cultured zebrafish follicles, activin A and inhibin A both induced final oocyte maturation in a dose-dependent manner. The effects of activin A and inhibin A were blocked by their binding protein, follistatin-288. Interestingly, follistatin-288 also inhibited final oocyte maturation induced by gonadotropin and by maturation inducing hormone (MIH), suggesting that activin A and/or inhibin A may be local regulators mediating gonadotropin- and MIH-induced final oocyte maturation. Taken together, these findings suggest that activin A and inhibin A are paracrine regulators of ovarian functions in fish. PMID- 10819760 TI - Identification of INSL6, a new member of the insulin family that is expressed in the testis of the human and rat. AB - A new member of the insulin gene family (INSL6) was identified from an Expressed Sequence Tag database through a search for proteins containing the insulin family B-chain cysteine motif. Human and rat INSL6 encoded polypeptides of 213 and 188 amino acids, respectively. These orthologous sequences contained the B-chain, C peptide, and A-chain motif found in other members of the insulin family. Human INSL6 was 43% identical to human relaxin H2 in the B- and A-chain regions. As with other family members, human and rat INSL6 had predicted dibasic sequences at the junction of the C-peptide and A-chain. Human INSL6 sequence had an additional dibasic site near the C-terminus of the A-chain. The presence of a single basic residue at the predicted junction of the B-chain and C-peptide suggests that multiple prohormone convertases are required to produce the fully mature hormone. INSL6 was found to be expressed at high levels in the testis as determined by Northern blot analysis and specifically within the seminiferous tubules in spermatocytes and round spermatids as detected by in situ hybridization analysis. Radiation hybrid mapping placed the human INSL6 locus at chromosome 9p24 near the placenta insulin-like homologue INSL4 and the autosomal testis-determining factor (TDFA) locus. PMID- 10819761 TI - Expression and action of kit ligand/stem cell factor in normal human and bovine ovarian surface epithelium and ovarian cancer. AB - Greater than 95% of ovarian cancers originate from the epithelial cells on the surface of the ovary termed ovarian surface epithelium (OSE). A normal aspect of OSE function is repeated proliferation after ovulation, and this is postulated to be involved in part in the onset of ovarian cancer. The hypothesis tested is that locally produced growth factors have an important role in controlling OSE proliferation. The current study investigates the potential role of the growth factor kit ligand (KL)/stem cell growth factor and its receptor c-kit in normal OSE biology and ovarian cancer. Human tumors from borderline, stage I, and stage III cases of ovarian cancer were found to express KL and c-kit protein in the epithelial cell component by ICC analysis. The stromal cell component of human ovarian tumors contained little immunostaining. Bovine ovarian physiology and endocrinology are similar to the human such that cow ovaries were used as a model system to investigate normal OSE functions. KL and c-kit proteins were detected in the OSE from both normal human and bovine ovaries. Adjacent ovarian stromal tissue contained less intense but positive KL and c-kit immunostaining. To extend the ICC results, RNA was collected from normal bovine OSE and ovarian stromal cells to examine KL gene expression. KL transcripts were detected in cultured OSE and stromal cells by Northern blot analysis. KL gene expression was found to be high in freshly isolated OSE but low in freshly isolated stroma using a quantitative polymerase chain reaction procedure. Levels of KL gene expression in cultured OSE and stroma increased to high levels. Observations indicate that normal OSE expresses high levels of KL in vivo and in vitro. The actions of KL on the growth of both normal OSE cells and ovarian cancer cells was investigated. KL was found to stimulate the growth of normal OSE cells in a similar manner to epidermal growth factor. Observations demonstrate the production and action of KL by normal OSE cells and ovarian cancer cells. Coexpression of KL and c-kit by normal OSE suggests that KL can act as an autocrine factor for OSE. The local production and action of KL on OSE provides insight into normal OSE biology, and a factor that may be involved in the onset and progression of ovarian cancer. PMID- 10819762 TI - Detection of progesterone receptor transcript in human spermatozoa. AB - The present study, to our knowledge, is the first to demonstrate presence of progesterone receptor (PR) transcript in human spermatozoa. The study shows the presence of low copy number PR mRNA in mature human spermatozoa. The PR transcript in spermatozoa was detected by reverse transcriptase-polymerase chain reaction using primers specific for the hormone binding domain and the DNA binding domain of the conventional uterine PR. Further, the cDNA sequence of the partial PR transcript from spermatozoa was found to be identical to the region spanning nucleotides 2694 to 3230 of the conventional PR full-length cDNA sequence. This study also indirectly suggests that the PR protein indeed is an intrinsic sperm protein and is not acquired through proteinaceous secretions of accessory reproductive organs. PMID- 10819763 TI - Protein tyrosine phosphorylation during prolonged in vitro incubation of ejaculated bovine spermatozoa is regulated by the oxidative state of the medium. AB - Protein tyrosine phosphorylation plays a regulatory role in a multitude of physiological processes in sperm. Changes in protein tyrosine phosphorylation, viability, and motility were studied as a function of extended incubation of bovine sperm in vitro at ambient temperature (18-20 degrees C). Fresh ejaculates were incubated after dilution for 8 days. On Days 0, 2, 5, and 8, an aliquot of sperm was incubated with or without theophylline at 37 degrees C for 30 min prior to assessing sperm viability, motility, and tyrosine phosphorylation of soluble and whole-cell proteins. There was a time-dependent decline in sperm motility, which was to some extent reversed by incubation with theophylline. The sum of the phosphotyrosine signal from two soluble proteins (M(r) 67 000 and 36 000) declined with incubation time in both theophylline-treated and untreated sperm. There were major differences in the pattern of tyrosine phosphorylation during incubation between ejaculates from different bulls. Tyrosine phosphorylation of a number of proteins from whole-cell extracts increased in a time-dependent manner during in vitro incubation and was unaffected by the presence of theophylline in the medium. The oxygenation state of the incubation medium had profound effects on sperm motility, viability, and tyrosine phosphorylation of proteins from whole cell extracts. Sperm motility and viability declined more rapidly under aerobic compared with anaerobic conditions. Tyrosine phosphorylation of proteins from whole-cell extracts increased considerably during anaerobic incubation, while there was no significant change during aerobic incubation. This increase in phosphorylation due to anaerobic incubation was reversed when sperm were transferred from an anaerobic to an aerobic environment, indicating that the oxygenation state of the medium regulates both protein tyrosine kinases and phosphatases. In addition, sperm incubated under aerobic conditions for 5 days retained the ability to phosphorylate proteins when transferred to an anaerobic environment. The increase in protein tyrosine phosphorylation during in vitro incubation took place in a medium that did not contain capacitating substances such as heparin, sodium bicarbonate, or BSA. It transpired over a time scale of days and was not augmented by an increase in intracellular cAMP concentration through phosphodiesterase inhibition. Protein tyrosine phosphorylation during extended in vitro incubation at ambient temperature was significantly inhibited by the presence of oxygen in the medium. PMID- 10819764 TI - Pregnancy-associated bovine and ovine glycoproteins exhibit spatially and temporally distinct expression patterns during pregnancy. AB - The pregnancy-associated glycoproteins (PAG) constitute a large family of recently duplicated genes. They show structural resemblance to pepsin and related aspartic proteinases. A total of 21 bovine (bo) PAG and 9 ovine (ov) PAG cDNA have been identified. Phylogenetic analysis indicated that the PAG are divided into two main groupings that accurately reflect their tissue expression, as determined by in situ hybridization. In the first pattern, represented by ovPAG-2 and boPAG-2, -8, -10, and -11 (where the numbering is arbitrary and reflects order of discovery within species), expression occurred throughout the outer epithelial layer of the placenta (trophectoderm). The second pattern was predominant localization to binucleate cells. Ribonuclease protection assays, which allow discrimination between closely related transcripts, have shown that the expression of PAG varies in a temporal manner over pregnancy. Of those bovine PAG expressed predominantly in binucleate cells, boPAG-1, -6, and -7 are expressed weakly, if at all, by Day 25 placenta, but are present at the middle and end of pregnancy. Others, such as boPAG-4, -5, and -9, are expressed at Day 25 and at earlier stages. Although not among the earliest PAG produced by the trophoblast, boPAG-1 has been used for pregnancy diagnosis, particularly in dairy cows, where there is a major need for a sensitive method capable of detecting pregnancy within 1 mo of conception. It seems likely that some of the newly discovered PAG will be better candidates than PAG-1 for pregnancy diagnosis. PMID- 10819766 TI - Establishment of a porcine cell line from in vitro-produced blastocysts and transfer of the cells into enucleated oocytes. AB - The present study was conducted to establish a porcine cell line from blastocysts produced in vitro and to examine the developmental ability of nuclear transfer embryos reconstituted with the cells and enucleated mature oocytes. When hatched blastocysts were cultured in Dulbecco's modified Eagle's medium with supplements, no colonies of embryo-derived cells were observed. In contrast, 56% of embryos that were attached to feeder layers of STO cells formed colonies in NCSU-23 with supplements. When the colonies were subcultured in the absence of feeder cells, a cell line with an epithelial-like cell morphology was obtained. This cell morphology was stable up to at least passage 30. Although no fused embryos were observed when a pulse of 100 V/mm was applied, the fusion rate increased significantly at 150 V/mm (28%) and 200 V/mm (64%). At 200 V/mm, 39% of fused embryos cleaved, but no embryos developed beyond the 3-cell stage. When cocultured with electro-activated oocytes, percentages of reconstructed embryos cleaved (65%) and developed to the 4-cell stage (23%) were significantly higher than percentages for those (cleavage: 38%; 4-cell stage: 3%) in the absence of activated oocytes. At 7 days after culture, one reconstructed embryo successfully developed to the blastocyst stage in the presence of activated oocytes. When green fluorescent protein-expressing cells and enucleated oocytes were fused and the fused embryos were cultured with electro-activated oocytes, 3 of 102 reconstructed embryos developed to the blastocyst stage. All of the blastocysts were positive for fluorescent green under ultraviolet light. The results of the present study indicate that a porcine cell line can be established from the hatched blastocyst and maintained in vitro for a long period, and that reconstructed embryos obtained by transferring the blastocyst-derived cells into enucleated oocytes have the ability to develop to the blastocyst stage in vitro. PMID- 10819765 TI - Macrophage migration inhibitory factor-induced Ca(2+) response in rat testicular peritubular cells. AB - Macrophage migration inhibitory factor (MIF), originally described as a T-cell product, has recently been identified in several endocrine organs. In the rat testis, MIF is secreted by the Leydig cells into testicular interstitial fluid that directly contacts Sertoli and peritubular cells. To investigate whether MIF is involved in calcium-dependent signal transduction, we have isolated rat Sertoli and peritubular cells. Despite progress in understanding functional properties of MIF, the molecular mechanism of MIF action in target cells is almost completely unknown. Here we find that recombinant MIF evokes a transient increase in calcium levels in peritubular cells but not in Sertoli cells from dissociated rat testis. Concentrations in the range between 12.5 ng/ml and 120 ng/ml of recombinant MIF were found to be effective, with 50 ng/ml yielding the largest increase in intracellular calcium. Preincubation of MIF with a neutralizing monoclonal antibody specifically blocked the response. Incubation of the peritubular cells in calcium-free buffer clearly decreased the evoked response in intracellular calcium concentration. However, the calcium response was greatly decreased by thapsigargin, an inhibitor of the Ca(2+) ATPase of the endoplasmic reticulum. The results strongly indicate that calcium is mobilized from reticulum stores during MIF-mediated signal transduction in the testis. In conclusion, our results provide the first characterization of MIF signal transduction in the testis and suggest that signaling from Leydig cells to peritubular cells through MIF is mediated by receptors coupled to release of intracellular calcium. PMID- 10819767 TI - A role for phosphorylation of glycogen synthase kinase-3alpha in bovine sperm motility regulation. AB - The long-term goal of our work is to understand biochemical mechanisms underlying sperm motility and fertility. In a recent study we showed that tyrosine phosphorylation of a 55-kDa protein varied in direct proportion to motility. Tyrosine phosphorylation of the protein was low in immotile compared to motile epididymal sperm. Inhibition or stimulation of motility by high calcium levels or cAMP, respectively, results in a corresponding decrease or increase in tyrosine phosphorylation of the 55-kDa protein. Here we report purification and identification of this motility-associated protein. Soluble extracts from bovine caudal epididymal sperm were subjected to DEAE-cellulose, Affi-Gel blue, and cellulose phosphate chromatography. Tyrosine phosphate immunoreactive fractions contained glycogen synthase kinase-3 (GSK-3) activity, suggesting a possible correspondence between these proteins. This suggestion was verified by Western blot analyses following one-dimensional and two-dimensional gel electrophoresis of the purified protein using monoclonal and affinity-purified polyclonal antibodies against the catalytic amino-terminus and carboxy-terminus regions of GSK-3. Further confirmation of the identity of these proteins came from Western blot analysis using antibodies specific to the tyrosine phosphorylated GSK-3. Using this antibody, we also showed that GSK-3 tyrosine phosphorylation was high in motile compared to immotile sperm. Immunocytochemistry revealed that GSK-3 is present in the flagellum and the anterior portion of the sperm head. These data suggest that GSK-3, regulated by phosphorylation, could be a key element underlying motility initiation in the epididymis and regulation of mature sperm function. PMID- 10819768 TI - Association of the mouse infertility factor DAZL1 with actively translating polyribosomes. AB - The DAZ (Deleted in AZoospermia) gene family was isolated from a region of the human Y chromosome long arm that is deleted in about 10% of infertile men with idiopathic azoospermia. DAZ and an autosomal DAZ-like gene, DAZL1, are expressed in germ cells only. They encode proteins with an RNA recognition motif and with either a single copy (in DAZL1) or multiple copies (in DAZ) of a DAZ repeat. A role for DAZL1 and DAZ in spermatogenesis is supported by their homology to a Drosophila male infertility protein Boule and by sterility of Dazl1 knock-out mice. The biological function of these proteins remains unknown. We found that DAZL1 and DAZ bound similarly to various RNA homopolymers in vitro. We also used an antibody against the human DAZL1 to determine the subcellular localization of DAZL1 in mouse testis. The sedimentation profiles of DAZL1 in sucrose gradients indicate that DAZL1 is associated with polyribosomes, and further capture of DAZL1 on oligo(dT) beads demonstrates that the association is mediated through the binding of DAZL1 to poly(A) RNA. Our results suggest that DAZL1 is involved in germ-cell specific regulation of mRNA translation. PMID- 10819769 TI - Hormonal variation of rat uterine contractile responsiveness to selective neurokinin receptor agonists. AB - Regulated uterine contractions are important in many reproductive functions such as sperm transport and embryo positioning during implantation. The role of classical neurotransmitters including acetylcholine and norepinephrine in regulating myometrial contractility has been well studied; however, the peripheral role of sensory neurotransmitters such as the neurokinins is less clear. The major neurokinins are substance P, neurokinin A, and neurokinin B, which predominantly activate neurokinin receptors (NK-Rs) 1, 2, and 3, respectively. This study utilized selective receptor agonists to examine the role of NK-Rs in uterine contractility. Uterine tissues, obtained from the major stages of the rat estrous cycle, were stimulated with selective NK-R agonists. Addition of each agonist resulted in a significant contractile response. However, the magnitude and nature of the response were dependent upon the stage of the estrous cycle, with responses to all agonists being significantly decreased in tissue from proestrus and estrus. Furthermore, the nature of NK3-R-mediated contraction was different in tissue from proestrus and estrus compared to metestrus and diestrus. The hormonal dependence of NK-R-mediated contractility was then examined in the ovariectomized estrogen-supplemented rat model. These studies confirmed that the magnitude and nature of uterine contractility in response to NK-R activation depend upon the hormonal environment. PMID- 10819770 TI - Rat sperm 2B1 glycoprotein (PH20) contains a C-terminal sequence motif for attachment of a glycosyl phosphatidylinositol anchor. Effects of endoproteolytic cleavage on hyaluronidase activity. AB - Rat sperm 2B1 antigen (the orthologue of guinea pig sperm PH20) is a plasma membrane-bound glycoprotein that is endoproteolytically cleaved during passage through the epididymis and subsequently migrates from the tail to the acrosomal domain during capacitation. Unlike guinea pig PH20, however, sperm surface 2B1 is insensitive to phosphatidylinositol phospholipase C, nor is it known how endoproteolytic cleavage affects its hyaluronidase activity. In this investigation we have expressed 2B1 cDNA in Chinese hamster ovary cells; we have shown that it contains an internal sequence motif for attachment of a glycosyl phosphatidylinositol (GPI) anchor and that cleavage from a single- into a two chain molecule causes a significant shift in the optimum pH for hyaluronidase activity. Functionally, these results suggest that 1) 2B1 glycoprotein on rat spermatozoa is attached to the plasma membrane via a GPI anchor and that this is an important factor in its ability to migrate from the tail to the acrosomal domain during capacitation; and 2) endoproteolytic cleavage of 2B1 serves to optimize its hyaluronidase activity immediately before fertilization, thereby facilitating penetration of spermatozoa through the cumulus oophorus. PMID- 10819771 TI - Onset of the first S-phase is determined by a paternal effect during the G1-phase in bovine zygotes. AB - The aim of this study was to characterize the respective influences of the paternal and the maternal components on the timing of the first S-phase in the bovine zygote. In vitro-matured oocytes were fertilized in vitro with sperm conferring a high blastocyst rate (embryos of group 1) or a low blastocyst rate (embryos of group 2). Resulting zygotes were either allowed to develop in vitro to the blastocyst stage or exposed to 5'-bromo-2'-deoxyuridine in order to characterize the timing of their first S-phases. Timing of pronuclear formation was similar in the two groups, but the onset of S-phase and the first cleavage occurred earlier in group 1 than in group 2. We also showed that the length of the S-phase represented 30% of the first cell cycle in group 1 and 20% in group 2. Differences in times of onset of the first S-phase observed between embryo groups concerned both male and female pronuclei in a similar manner and were not dependent on the maternal component of the zygote. Our data demonstrated that the precocity of the onset of the first S-phase stemmed from a paternal control exerted during a transient period of the G1-phase. PMID- 10819772 TI - Effect of neonatal gonadotropin-releasing hormone antagonist administration on sertoli cell number and testicular development in the marmoset: comparison with the rat. AB - The primary purpose of this study was to establish whether Sertoli cells proliferate in the neonatal period in the marmoset monkey (Callithrix jacchus) and whether administration of a long-acting GnRH antagonist (GnRHa) during this phase induced any transient or permanent effects on Sertoli cell number or on any other aspect of testicular development. Male marmoset co-twins (n = 9) were treated during Weeks 1-14 with either vehicle or GnRHa. Four sets of co-twins were examined at Weeks 18-22 (start of infancy) and 5 sets in adulthood (92+ wk), and Sertoli cell number was determined using either the nucleator or optical disector methods; other testicular morphometric analyses (e.g., germ cell volume, Leydig cell volume) used standard point-counting. Data for the marmoset were compared with that obtained in similarly treated rats. Sertoli cell number in marmosets treated neonatally with GnRHa was reduced by 35% compared with that of controls at Weeks 18-22 but was comparable to control values in adulthood. However, seminiferous epithelium volume was reduced significantly in adult marmosets treated neonatally with GnRHa, and there was a tendency for reduced germ cell volume per Sertoli cell. In the same animals, there was significant expansion of the interstitium and an increase in Leydig cell volume per testis when compared with co-twin controls; a similar increase in Leydig cell volume was evident in adult rats treated neonatally with GnRHa. Comparison of Sertoli cell numbers in 6 infantile (18-24 wk) and 10 adult marmosets showed that adult numbers of Sertoli cells were present by the start of infancy but, unlike rats, marmosets were still able to replicate Sertoli cells beyond this period. However, marmoset Sertoli cells supported only approximately 20% of the germ cell volume supported by rat Sertoli cells, indicative of poor efficiency of spermatogenesis, as shown previously in the human. This finding, together with the demonstration of a temporal pattern of Sertoli cell replication similar to that in the human, supports the use of marmosets as a model for human male testicular development and function. PMID- 10819773 TI - Molecular cloning and characterization of phosphatidylcholine transfer protein like protein gene expressed in murine haploid germ cells. AB - We have isolated a cDNA clone specifically expressed in spermiogenesis from a subtracted cDNA library of mouse testis. The cDNA consisted of 1392 nucleotides and had an open reading frame of 873 nucleotides encoding a protein of 291 amino acid residues. Computer-mediated homology search revealed that the nucleotide sequence was unique but the deduced amino acid sequence had similarity to mouse phosphatidylcholine transfer protein (PCTP). We named this newly isolated gene PCTP-like protein. Northern blot analysis revealed a 1.4-kilobase mRNA expressed in the testis, kidney, liver, and intestine with the highest level in the testis. Messenger RNA expression in the testis was detected first on Day 23 in postnatal development and then increased up to adulthood. The protein, having a molecular weight of approximately 40 000, was encoded by the mRNA and was detected at the tail of the elongated spermatids and sperm by immunohistochemical staining. PMID- 10819774 TI - Ability of integrins to mediate fertilization, intracellular calcium release, and parthenogenetic development in bovine oocytes. AB - The ability of arginine-glycine-aspartic acid (RGD; a sequence recognized by integrins) or non-RGD-containing peptides to block fertilization, induce intracellular Ca(2+) oscillations, and initiate parthenogenetic development in bovine oocytes was investigated. Addition of a soluble RGD peptide during fertilization at concentrations ranging from 10 to 1000 microg/ml significantly decreased (P<0.05) fertilization as compared to the in vitro-fertilized controls. The addition of non-RGD peptide had no effect on fertilization. Two intracellular Ca(2+) transients 21.5+/- 1.9 min apart were observed in 56 of 60 oocytes incubated in RGD peptide concentrations ranging from 20 to 1000 microg/ml. No intracellular Ca(2+) transients were observed in medium alone, non-RGD treatment groups or in the RGD peptide at 10 microg/ml. The percentage of oocytes activated with ionomycin and 6-dimethylaminopurine (63% cleavage and 34% blastocyst development) was significantly higher (P<0.05) than those activated with the RGD peptide and 6-dimethylaminopurine (35% cleavage and 19% blastocyst development). These groups were significantly higher (P<0.05) than either peptide alone, 6 dimethylaminopurine alone, or the non-RGD peptide and 6-dimethylaminopurine treatment groups. These data provide evidence that ligation of an integrin on bovine oocytes with a soluble RGD peptide is capable of blocking fertilization, inducing intracellular Ca(2+) transients, and initiating parthenogenetic development. PMID- 10819775 TI - Estrogen-astrocyte-luteinizing hormone-releasing hormone signaling: a role for transforming growth factor-beta(1). AB - The purpose of this study was to identify factors from astrocytes that can regulate LHRH neurosecretion. Exposure of LHRH-secreting (GT1-7) cells to conditioned media (CM) from C6 glial cells and hypothalamic astrocytes (HA) stimulated LHRH release. Assays of C6 and HA CM revealed that transforming growth factor-beta(1) (TGF-beta(1)) and 3alpha-hydroxy-5alpha-pregnane-20-one (3alpha, 5alpha-THP), both known LHRH secretagogues, were present in CM and their levels increased in parallel to the LHRH-releasing activity of CM. In contrast, TGF alpha was undetectable in C6 or HA CM. Ultrafiltration to remove peptides with molecular weights >10 kDa virtually abolished the LHRH-releasing ability of the HA CM. Furthermore, immunoneutralization with a panspecific THF-beta antibody dose-dependently attenuated the LHRH-releasing activity of the CM. Rat hypothalamus and GT1-7 cells were demonstrated to express TGF-beta receptors as well as furin, an enzyme that converts latent TGF-beta(1) to active TGF-beta(1). Estrogen receptor-alpha and ER-beta mRNA and protein were also demonstrated in HAs by reverse transcription-polymerase chain reaction and double immunofluorescence, and treatment with 17beta-estradiol (17beta-E(2)) increased both active and latent TGF-beta(1) levels in HA CM. The effect of 17beta-E(2) was completely blocked by the ER antagonist ICI8280. As a whole, these studies provide evidence of a previously undescribed 17beta-E(2)-TGF-beta(1)-LHRH signaling pathway. PMID- 10819776 TI - Sexual dimorphism in the regulation of meiotic process in the rabbit. AB - Meiosis, mitosis, and apoptosis during fetal and postnatal periods were investigated in order to explore mechanisms of sexual dimorphism in initiation of germ cell meiosis. Gonads were obtained from Japanese white rabbits from 23 to 51 days postcoitum (dpc). Gonadal thin sections were stained with hematoxylin and eosin. Germ cell alkaline phosphatase and apoptosis were detected with histochemical and immunohistochemical methods, respectively. In the ovary, meiotic germ cells were initially recognized at 29 dpc and arrested after enclosure within follicles. Similarly, meiotic germ cells were recognized outside seminiferous tubules at 29 dpc, but no meiotic figures were identified in intratubular spaces. Apoptotic germ cells were not recognized in the intratubular spaces before 35 dpc, and no apoptotic figures were recognized in the ovary during the period studied. In conclusion, the initiation of meiosis in testicular interstitial tissue at the time comparable to that in the ovary indicates that germ cells of both sexes have the ability to enter meiosis during the same stage of fetal development; and it appears most likely that delayed initiation of meiosis in the intratubular space is attributable to meiosis-inhibiting substance(s) present in seminiferous tubules. PMID- 10819777 TI - Interferon-tau stimulates granulocyte-macrophage colony-stimulating factor gene expression in bovine lymphocytes and endometrial stromal cells. AB - Interferon-tau (IFN-tau), the antiluteolytic signal produced by the trophoblast prior to implantation in ruminants, exhibits immunomodulatory properties. It stimulates the production of prostaglandin (PG) E(2) in bovine endometrial cells via the induction of cyclooxygenase-2 (COX-2). We previously demonstrated that preconditioning lymphocytes with PGE(2) increases the expression of granulocyte macrophage colony-stimulating factor (GM-CSF), a cytokine that promotes conceptus growth and survival. Our goal in the present study was to evaluate the impact of IFN-tau on the expression of GM-CSF in bovine peripheral blood lymphocytes (PBL) and endometrial epithelial and stromal cells. Changes in PGE(2) production and mRNA levels of COX-2 were also studied in PBL in response to IFN-tau. Gene expression was estimated by semiquantitative reverse transcription-polymerase chain reaction and Northern analysis. The expression of GM-CSF in PBL was stimulated by treatment with IFN-tau. Furthermore, GM-CSF mRNA levels were increased after preconditioning PBL for 3 days with IFN-tau, followed by a 12-h restimulation without IFN-tau. Inhibition rather than stimulation of PGE(2) production and COX-2 expression in PBL during treatment with IFN-tau suggests a direct effect on GM-CSF expression. Moreover, GM-CSF expression was stimulated in uterine stromal cells in response to IFN-tau. This study provides the first evidence for stimulation of GM-CSF expression by IFN-tau in both leukocytes and endometrial stromal cells. In view of the role of GM-CSF on fetal growth and survival, these results support the hypothesis that the conceptus mediates accommodation mechanisms in the uterus during early pregnancy by modulating the expression of beneficial cytokines at the fetomaternal interface. PMID- 10819778 TI - Significance of the number of embryonic cells and the state of the zona pellucida for hatching of mouse blastocysts in vitro versus in vivo. AB - We investigated the course of mouse blastocyst hatching in vitro after experimental modulation of the hatching process by growth hormone or by laser treatment and compared it to embryos grown in vivo. When embryos were grown in vitro, successful hatching was dependent on blastocyst expansion and was based on a minimum number of embryonic cells. Embryos grown in the presence of growth hormone were more advanced in their development and hatched earlier. When an artificial opening was laser-drilled into the zona pellucida, hatching occurred at lower numbers of embryonic cells. In vivo, escape from the zona pellucida occurred earlier and independent of blastocyst expansion. However, when we isolated in vivo-grown blastocysts with intact zonae that had developed in vivo and then cultured them in vitro, blastocysts started to expand and hatched the following day when a sufficiently high number of embryonic cells was present. Our data show that successful hatching in vitro is dependent on a sufficiently high number of embryonic cells, which enables blastocyst expansion and zona shedding. In vivo, the lower number of embryonic cells detected in zona-free blastocysts indicates that the underlying mechanism of zona escape is different, does not depend on blastocyst expansion, and presumably involves lytic factors from the uterus. PMID- 10819779 TI - Involvement of mitochondria in oxidative stress-induced cell death in mouse zygotes. AB - Accumulation of reactive oxygen species during aging leads to programmed cell death (PCD) in many cell types but has not been explored in mammalian fertilized eggs, in which mitochondria are "immature," in contrast to "mature" mitochondria in somatic cells. We characterized PCD in mouse zygotes induced by either intensive (1 mM for 1.5 h) or mild (200 microM for 15 min) hydrogen peroxide (H(2)O(2)) treatment. Shortly after intensive treatment, zygotes displayed PCD, typified by cell shrinkage, cytochrome c release from mitochondria, and caspase activation, then terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining in condensed pronuclei. On the other hand, after mild treatment, zygotes arrested developmentally and showed neither cytochrome c release nor caspase activation over 48 h; until 72 h, 46% zygotes exhibited TUNEL staining, and 88% of zygotes lost plasma membrane integrity. Interestingly, mild oxidative treatment induced a decline in mitochondrial membrane potential and disruption of the mitochondrial matrix. Taken together, these results suggest that oxidative stress caused by H(2)O(2) induces PCD in mouse zygotes and that mitochondria are involved in the early phase of oxidative stress-induced PCD. Furthermore, mitochondrial malfunction also may contribute to cell cycle arrest, followed by cell death, triggered by mild oxidative stress. PMID- 10819780 TI - In vitro-cultured bovine oviductal cells bind acrosome-intact sperm and retain this ability upon sperm release. AB - The mammalian oviduct plays a key role in sperm storage, capacitation, and selection. Specific oviduct secretions and/or binding to oviductal cells are thought to be responsible for the extension of the fertile life span of sperm. In this in vitro study, a quantitative assay for sperm binding was developed to analyze the mechanisms of sperm-oviductal cell adhesion and release in the bovine species. Distribution and acrosomal status of sperm bound to in vitro-cultured ampullary and isthmic cell monolayers were followed until the time of sperm release by means of fluorescence labeling techniques. In order to understand whether release is due to surface changes of sperm or oviductal cells, double incubation experiments with unlabeled and Hoechst-labeled sperm have been performed. Main findings demonstrate that (1) only acrosome-intact sperm bind specific bovine oviductal epithelial cells; (2) acrosomes of bound sperm are preserved intact over time; and (3) release of unreacted sperm is likely to be due to changes of the sperm surface, probably triggered by capacitation. These findings support the hypothesis that binding to oviductal cells is essential for preserving the sperm fertilization competence during the interval from the onset of estrus to ovulation. PMID- 10819781 TI - Transcriptional interference between glucocorticoid receptor and estradiol receptor mediates the inhibitory effect of cortisol on fish vitellogenesis. AB - In oviparous species, the synthesis of vitellogenin (Vg) takes place in the liver according to a strictly estrogen-dependent mechanism that first involves an up regulation of the estrogen receptor (ER) by its own ligand. However, reports from the literature indicate that in trout stress or cortisol may cause a reduction of cytosolic E2-binding sites in the liver and a decrease in plasma Vg levels. To investigate the mechanisms underlying these effects, in vivo and in vitro experiments were designed in rainbow trout (Oncorhynchus mykiss). The results demonstrate that cortisol implanted into maturing females caused a marked decrease of rainbow trout ER (rtER) and rainbow trout Vg (rtVg) mRNA levels in the liver. In vitro experiments on hepatocyte aggregates also showed that dexamethasone (Dex) caused a strong decrease in the basal and E2-stimulated rtER mRNA and to a lesser extent rtVg mRNA. These effects were specific as no other hormones were able to mimic the inhibitory action of Dex. A study of rtER mRNA stability indicated that the effects of glucocorticoids are likely to take place at the transcriptional level. This was further indicated by transfection experiments in CHO-K(1) cells, which showed that rainbow trout glucocorticoid receptor (rtGR) strongly inhibited the E2-stimulated transcriptional activity of the rtER promoter. Taken together, these results indicate that the rtGR exerts a transcriptional interference on the expression of the rtER that may explain some of the negative effects of stress or cortisol on vitellogenesis. PMID- 10819782 TI - Keratinocyte growth factor: expression by endometrial epithelia of the porcine uterus. AB - Keratinocyte growth factor/fibroblast growth factor-7 (KGF/FGF-7) is an established paracrine mediator of hormone-regulated epithelial growth and differentiation. In all organs studied, KGF is uniquely expressed in cells of mesenchymal origin. To determine whether KGF and its receptor, keratinocyte growth factor receptor (KGFR) or fibroblast growth factor receptor-2IIIb, were expressed in the porcine uterus as a potential paracrine system mediating progesterone action, we cloned KGF and KGFR partial cDNAs from the porcine endometrium. KGF and KGFR expression was detected in endometrium by Northern blot hybridization. Interestingly, in situ hybridization results demonstrated that KGF was expressed by endometrial epithelia and was particularly abundant between Days 12 and 15 of the estrous cycle and pregnancy. KGF secretion into the lumen of the porcine uterus was also detected on Day 12 of the estrous cycle and pregnancy. KGFR was expressed in both endometrial epithelia and conceptus trophectoderm. These novel findings suggest that KGF may act on the uterine endometrial epithelium in an autocrine manner and on the conceptus trophectoderm in a paracrine manner in the pig, which is the only species possessing a true epitheliochorial type of placentation. PMID- 10819783 TI - Prolactin receptor and uterine milk protein expression in the ovine endometrium during the estrous cycle and pregnancy. AB - Lactogenic hormones regulate epithelial proliferation, differentiation, and function in a variety of epitheliomesenchymal organs. During pregnancy, the ovine uterus is a potential site for endocrine and paracrine actions of lactogenic hormones in the form of pituitary prolactin (PRL) and placental lactogen (PL). These studies determined temporal and spatial alterations in PRL receptor (PRL-R) and expression of uterine milk proteins (UTMP), a marker of endometrial secretory activity, in the ovine endometrium during the estrous cycle and pregnancy. Slot blot hybridization analysis indicated that steady-state levels of endometrial PRL R mRNA increased during pregnancy. In situ hybridization and immunohistochemical analyses indicated that PRL-R mRNA and protein were exclusively expressed in the endometrial glandular epithelium (GE). No PRL-R mRNA expression was detected in luminal epithelium, stroma, myometrium, or conceptus trophectoderm. Reverse transcription-polymerase chain reaction analyses determined that the endometrial GE expressed both long and short alternative splice forms of the ovine PRL-R gene. Slot-blot hybridization analysis indicated that steady-state levels of intercaruncular endometrial UTMP mRNA increased about 3-fold between Days 20 and 60, increased another 3-fold between Days 60 and 80, and then declined slightly to Day 120. In pregnant ewes, UTMP mRNA expression was restricted to the endometrial GE in the stratum spongiosum (sGE), increased substantially between Days 15 and 17, and, between Days 17 to 50 of gestation, was markedly higher in upper than lower sGE. After Day 50, hyperplasia of the sGE was accompanied by increased UTMP mRNA expression by all sGE. Collectively, results indicate that 1) endometrial sGE is a primary target for actions of lactogenic hormones and 2) UTMP mRNA expression is correlated with PL production by the trophectoderm and state of sGE differentiation during pregnancy. It is proposed that activation of PRL-R signal transduction pathways by PRL and PL plays a major role in endometrial GE remodeling and differentiated function during pregnancy in support of conceptus growth and development. PMID- 10819784 TI - Stallion epididymal fluid proteome: qualitative and quantitative characterization; secretion and dynamic changes of major proteins. AB - Proteins present in and secreted into the lumen of various regions of the stallion epididymis were characterized qualitatively and quantitatively by two dimensional electrophoresis. Using this proteomic approach, 201 proteins were found in the lumen and 117 were found that were secreted by the epithelium in various parts of the organ. Eighteen proteins made up 92.6% of the total epididymal secretory activity, lactoferrin (41.2%) and clusterin (24.8%) being the most abundant. Procathepsin D, HE1/CTP (cholesterol transfer protein), GPX (glutathione peroxidase), beta-N-acetyl-hexosaminidase, and PGDS (prostaglandin D2 synthase) were the other major compounds secreted. The most abundant proteins found in the luminal fluid were albumin and the secreted proteins: lactoferrin, PGDS, GPX, HE1/CTP, and hexosaminidase. Three main secretory epididymal regions were identified from the protein pattern, i.e., regions E0-E2, E3-E5, and E6-E9. Region E0-E2 was characterized by the secretion of clusterin (53%), PGDS (44%), and GPX (6%). Region E3-E5 had the highest number of secreted proteins, the highest protein concentrations (60-80 mg/ml), and the highest spermatocrit value (85%). Lactoferrin (60% in E4), clusterin (29% in E3), hexosaminidase (10% in E3), and procathepsin D (6.9% in E4) were the most abundant proteins in this region. Region E6-E9, in which few region-specific secreted compounds were found, was characterized by a high quantity of lactoferrin in the luminal fluid (2-14 mg/ml). Comparison between the secretion of the major proteins and their concentrations in the lumen throughout the organ showed that the behavior of each protein is specific, in particular for the three isoforms of clusterin. PMID- 10819785 TI - Horse conceptuses secrete insulin-like growth factor-binding protein 3. AB - Insulin-like growth factor-I (IGF-I) promotes early embryonic development in several species. In the rabbit, IGF-I binds to the embryonic coats from Day 3 of development onward by a 38-kDa protein that is probably insulin-like growth factor-binding protein 3 (IGFBP3). In the present study, ligand, Western, and Northern blot analyses were used to demonstrate the presence of IGF-I-binding activity, several immunoreactive IGFBP3 proteins, and IGFBP3 mRNA in horse conceptuses with particularly large amounts of immunoreactive IGFBP3 in the conceptus capsule. In addition, immunoprecipitation of radiolabeled proteins showed that cultured horse conceptuses secreted IGFBP3 into the culture medium. Endometrial samples from mares also contained IGFBP3 mRNA and protein; but there was no evidence of secretion of IGFBP3 into the uterine lumen by ligand blot analysis, and there was evidence of only very small amounts by Western blot analysis. These results indicate that the horse conceptus secretes significant quantities of IGFBP3 toward the conceptus capsule from as early as Day 10 after ovulation. Thus, most of the IGFBP3 contained within the capsule, which binds IGF I to this special extracellular matrix of the preimplantation horse conceptus, is likely to be embryonic in origin. IGFBP3 in the horse conceptus capsule may enhance or modulate the action of IGFs on the developing conceptus. PMID- 10819786 TI - Cell type-specific regulation of fetal fibronectin expression in amnion: conservation of glucocorticoid responsiveness in human and nonhuman primates. AB - The appearance of oncofetal fibronectin (FFN) in cervical and vaginal secretions is predictive of human labor. Levels of FFN in amnion increase with the onset of labor in rhesus monkeys. Since glucocorticoid (GC) levels in serum and amniotic fluid increase in association with parturition, we compared GC-mediated regulation of FFN expression in cultures of amnion epithelial cells and fibroblasts isolated from human and baboon amnions. Cells were maintained with and without dexamethasone (DEX), and levels of FFN in the conditioned media were determined by ELISA. We observed that DEX treatment suppressed FFN levels in both human and baboon amnion epithelial cells, whereas it increased FFN levels in amnion fibroblasts. DEX treatment reduced FFN levels in cytotrophoblasts from human placenta and increased FFN levels in placental fibroblasts. Northern blots revealed that DEX reduced levels of fibronectin (FN) mRNA in amnion epithelial cells and cytotrophoblasts, whereas it increased FN mRNA in amnion and placental fibroblasts. We conclude that GC differentially regulates FFN expression in epithelial and mesenchymal cells from amnion and placenta. In addition, this pattern of cell type-specific FFN regulation by GC is conserved in human and nonhuman primates and may be responsible for parturition-dependent changes in FFN expression in gestational tissues. PMID- 10819787 TI - Region-specific regulation of cytochrome P450 aromatase messenger ribonucleic acid by androgen in brains of male rhesus monkeys. AB - We demonstrated previously that testosterone regulates aromatase activity in the anterior/dorsolateral hypothalamus of male rhesus macaques. To determine the level of the androgen effect, we developed a ribonuclease protection assay to study the effects of testosterone or dihydrotestosterone (DHT) on aromatase (P450(AROM)) mRNA in selected brain areas. Adult male rhesus monkeys were treated with testosterone or DHT. Steroids in serum were quantified by RIA. Fourteen brain regions were analyzed for P450(AROM) mRNA. Significant elevations of its message over controls (P<0.05) were found in the medial preoptic area/anterior hypothalamus of both androgen treatment groups and the medial basal hypothalamus of the testosterone-treated males. Other brain areas were not affected by androgen treatment. We conclude that testosterone and DHT regulate P450(AROM) mRNA in brain regions that mediate reproductive behaviors and gonadotropin release. The P450(AROM) mRNA of other brain areas is not androgen dependent. Brain-derived estrogens may also be important for maintaining neural circuitry in brain areas not related to reproduction. The control of P450(AROM) mRNA in these areas may differ from what we report here, but it is equally important to understand the function of in situ estrogen formation in these areas. PMID- 10819788 TI - Evidence for spontaneous postlactational estrus in gray short-tailed opossums (Monodelphis domestica). AB - Previous studies of the gray short-tailed opossum have shown that ovarian activity and estrus are induced by male pheromones, but we recently documented urogenital sinus (UGS) estrus in postlactational females despite their isolation from the male stimuli known to be associated with induced estrus. Body weights and UGS smears were collected after removal of pups in midlactation (19-37 days postpartum), after weaning (55-61 days postpartum), or after pheromone exposure. Estradiol was measured by RIA in plasma samples collected from dams during lactation, after separation from pups, and at estrus. Average days to UGS estrus from pup removal or initial pheromone exposure differed (P<0.05) only between the midlactation and pheromone exposure groups. Postlactational females showed a decrease in body weight from the time of pup removal or weaning to estrus, which contrasts with the increase seen in pheromonally stimulated females. Plasma estradiol was elevated at estrus in all groups, and females that were paired with males at postlactational estrus mated and produced litters. This study demonstrates that gray short-tailed opossums consistently experience estrus within 2 wk of weaning their young and that postlactational estrus appears to be hormonally and behaviorally equivalent to estrus induced by direct exposure to male pheromones. PMID- 10819789 TI - Cytoplasm mediates both development and oxidation-induced apoptotic cell death in mouse zygotes. AB - Eggs must be the major locus of reproductive aging in women, because donation of eggs from younger to middle-aged women abrogates the effects of age on fertility. Oxidative stress, mitochondrial dysfunction, and apoptosis are associated with senescence. To develop an animal model of egg senescence, we treated mouse zygotes with 175 microM H(2)O(2) that induced mitochondrial dysfunction and developmental arrest, followed by delayed cell death, consistent with apoptosis. We reconstructed zygotes with nuclei and cytoplasm from treated or untreated zygotes, then followed development and apoptotic cell death in the reconstituted embryos. Pronuclear exchange between untreated, normal zygotes served as nuclear transfer controls. Rates of cleavage and development to morula and blastocysts were significantly lower (P<0.01) in zygotes reconstituted from untreated pronuclei and H(2)O(2)-stressed cytoplasts than those of nuclear transfer controls. Instead, the arrested, reconstituted zygotes displayed TUNEL staining at a similar rate to that of H(2)O(2)-treated controls, suggesting that apoptotic potential could be transferred cytoplasmically. On the other hand, rates of cleavage and development to morula and blastocyst of the reconstituted zygotes, derived from stressed pronuclei and untreated cytoplasm, were significantly increased (P<0.05), compared to those of H(2)O(2)-treated, control zygotes, indicating that healthy cytoplasm could partly rescue pronuclei from oxidative stress. Although oxidation stressed both nuclei and cytoplasm, cytoplasm was more sensitive than nuclei to oxidative stress. It is suggested that cytoplasm, most likely mitochondria, plays a central role in mediating both development and apoptotic cell death induced by oxidative stress in mouse zygotes. PMID- 10819790 TI - Somatostatin inhibits follicle-stimulating hormone-induced adenylyl cyclase activity and proliferation in immature porcine Sertoli cell via sst2 receptor. AB - The potential involvement of somatostatin (SRIF) in testicular function was studied by using as a model primary cultures of purified immature porcine Sertoli cells. In the present report we show that Sertoli cells express mRNA for sst2 SRIF receptor and display SRIF-sensitive adenylyl cyclase. Sensitivity of adenylyl cyclase to SRIF and its analogues is compatible with the pharmacological profile of this receptor type. Relevant cAMP production is similarly inhibited by SRIF in both basal and stimulated (by gonadotropin FSH or by forskolin) conditions. Moreover, the observed SRIF actions on Sertoli cells require functional coupling of specific membrane receptors to adenylyl cyclase via Gi proteins because pertussis toxin prevents SRIF-dependent inhibition of adenylyl cyclase in either basal or FSH-stimulated conditions. Given the potent antiproliferative actions of SRIF in other cell types, we further assessed the possible SRIF-dependent modulation of [(3)H]thymidine incorporation by Sertoli cells. Our data point to SRIF-mediated inhibition of both basal and FSH stimulated [(3)H]thymidine uptake. This inhibition of Sertoli cell proliferation is, at least in basal conditions, also blocked by pertussis toxin pretreatment. Altogether, these data suggest that SRIF may play a role as an (local) inhibitor of FSH actions in testicular development. PMID- 10819791 TI - Expression of hepatocyte growth factor and its receptor c-met in the ovine uterus. AB - Epithelial-mesenchymal interactions (EMI) are necessary for epithelial cell proliferation, differentiation, and function in the uterus and are mediated, in part, by paracrine growth factors of stromal origin. The objective of this study was to determine if hepatocyte growth factor (HGF, scatter factor) and its receptor c-met were present in the ovine uterus and to characterize their temporal and spatial expression during the estrous cycle and pregnancy. Reverse transcription-polymerase chain reaction was used to clone partial cDNAs for ovine HGF and c-met from endometrial total RNA. Northern blot analysis of endometrial RNA revealed expression of a 6-kb mRNA for HGF and an 8-kb mRNA for c-met in ovine endometrium. In situ hybridization demonstrated that HGF mRNA was expressed by stromal cells of the endometrium, whereas c-met mRNA was localized exclusively to luminal and glandular epithelial cells. In the early conceptus, HGF mRNA was expressed by chorioallantoic mesenchyme, and c-met was expressed by trophectoderm. Steady-state levels of endometrial c-met mRNA increased after Day 9 in both cyclic and pregnant ewes. The HGF mRNA was expressed during both the estrous cycle and early pregnancy. Results indicate that HGF is a stromal-derived paracrine growth factor in the ovine uterus and placenta that is potentially involved in endometrial epithelial-stromal interactions and chorioallantoic stromal-trophectodermal interactions. In the ovine uterus, HGF may stimulate epithelial morphogenesis and differentiated function required for establishment and maintenance of pregnancy, conceptus implantation, and placentation. PMID- 10819792 TI - Modulation of estrogen production and 17beta-hydroxysteroid dehydrogenase-type 1, cytochrome P450 aromatase, c-met, and protein kinase Balpha messenger ribonucleic acid content in rat ovarian granulosa cells by hepatocyte growth factor and follicle-stimulating hormone. AB - Hepatocyte growth factor (HGF) suppresses FSH-dependent estradiol-17beta (E(2)) production in ovarian granulosa cells (GC). The mechanisms of action for HGF in GC are unknown; however, activation of the HGF receptor, c-Met, can induce c Akt/protein kinase B (PKB)-mediated signal transduction in nonovarian cells. Using immature rat GC, the present study investigated the effects of HGF within the estrogen biosynthetic pathway, concomitant with changes in c-Met and PKBalpha mRNA expression. Granulosa cells were incubated with androstenedione and FSH, HGF, and/or dibutyryl-cAMP (Bu(2)-cAMP). Follicle-stimulating hormone and Bu(2) cAMP each stimulated estrone (E(1)) and E(2) synthesis at 48 h. Hepatocyte growth factor suppressed FSH-dependent E(2), but not E(1), synthesis. Semiquantitative reverse transcription-polymerase chain reaction showed that HGF impaired FSH supported 17beta-hydroxysteroid dehydrogenase type-1 (17beta-HSD) and cytochrome P450 aromatase (P450arom) mRNA levels. Hepatocyte growth factor did not reduce E(2) synthesis or 17beta-HSD and P450arom mRNA expression in the presence of Bu(2)-cAMP at 48 h. The FSH and HGF each down-modulated c-Met mRNA accumulation, whereas Bu(2)-cAMP increased c-Met mRNA content. Between 0 and 48 h a biphasic change in PKBalpha mRNA content occurred with either FSH or HGF; however, PKBalpha mRNA accumulation was augmented by HGF. Collectively, results suggest that HGF can suppress E(2) production in GC by disrupting cAMP-dependent 17beta HSD and P450arom. Changes in c-Met and PKBalpha mRNA content provide a potential link between HGF signaling and the FSH-dependent mechanisms that control the steroidogenic differentiation of GC. PMID- 10819793 TI - Regulation of transforming growth factor-beta-receptor type I and type II messenger ribonucleic acid expression in the hamster ovary by gonadotropins and steroid hormones. AB - The hormonal regulation of ovarian transforming growth factor-beta (TGF-beta) type I receptor (TbetaRI) and TbetaRII messenger (mRNA) expression was evaluated using cyclic and hypophysectomized hamsters. Northern blot analysis revealed that three TbetaRI and one TbetaRII gene transcripts were expressed in the hamster ovary. Reverse transcription-polymerase chain reaction quantitation revealed that receptor mRNA was differentially expressed during the estrous cycle. Although, mRNA levels for both receptor types increased steadily up to Day 4:0900 h, a sharp decline occurred following the gonadotropin surge. In fact, receptor mRNA started declining by Day 4:1200 h, long before the gonadotropin surge; however, only TbetaRI mRNA levels recovered partially by 1500 h to fall again by 1600 h. Although hypophysectomy preferentially reduced TbetaRII mRNA levels, gonadotropins as well as ovarian steroids significantly induced TbetaRI and TbetaRII mRNA expression within 48 h and 24 h, respectively; 5alpha dihydrotesterone (DHT) induced only TbetaRII mRNA. The induction of ovarian TbetaRI and TbetaRII mRNA by estradiol-17beta() or progesterone was severely attenuated by dexamethasone. A marked increase in serum cortisol coincided with the periovulatory rise in serum gonadotropins. These results suggest that the increase in TGF-beta receptor mRNA expression correlates with gonadotropin induced ovarian follicular development during the estrous cycle. Moreover, receptor mRNA expression is critically and differentially regulated by gonadotropins as well as ovarian steroids. Most importantly, glucocorticoid appears to play a critical modulatory role in the temporal expression of receptor mRNA in the ovary, hence, controlling folliculogenesis. PMID- 10819794 TI - Oxidative phosphorylation-dependent and -independent oxygen consumption by individual preimplantation mouse embryos. AB - The self-referencing electrode technique was employed to noninvasively measure gradients of dissolved oxygen in the medium immediately surrounding developing mouse embryos and, thereby, characterized changes in oxygen consumption and utilization during development. A gradient of depleted oxygen surrounded each embryo and could be detected >50 microm from the embryo. Blastocysts depleted the surrounding medium of 0.6+/-0.1 microM of oxygen, whereas early cleavage stage embryos depleted the medium of only 0.3+/-0.1 microM of oxygen, suggesting a twofold increase in oxygen consumption at the blastocyst stage. Mitochondrial oxidative phosphorylation (OXPHOS) accounted for 60-70% of the oxygen consumed by blastocysts, while it accounted for only 30% of the total oxygen consumed by cleavage-stage embryos. The amount of oxygen consumed by non-OXPHOS mechanisms remained relatively constant throughout preimplantation development. By contrast, the amount of oxygen consumed by OXPHOS in blastocysts is greater than that consumed by OXPHOS in cleavage-stage embryos. The amount of oxygen consumed by one-cell embryos was modulated by the absence of pyruvate from the culture medium. Treatment of one-cell embryos and blastocysts with diamide, an agent known to induce cell death in embryos, resulted in a decline in oxygen consumption, such that the medium surrounding dying embryos was not as depleted of oxygen as that surrounding untreated control embryos. Together these results validate the self-referencing electrode technique for analyzing oxygen consumption and utilization by preimplantation embryos and demonstrate that changes in oxygen consumption accompany important physiological events, such as development, response to medium metabolites, or cell death. PMID- 10819795 TI - Distribution and localization of calmodulin-binding proteins in bull spermatozoa. AB - Previous studies from our laboratory have shown that a decrease in the calmodulin binding properties of a few sperm proteins occurs during the capacitation process, an effect associated with a decrease in intracellular calmodulin concentrations. Using biotinylated-calmodulin nitrocellulose overlay assay on protein extracts of subcellular fractions of bull spermatozoa, one of these proteins (p32) is detected in the flagellar-enriched fractions, whereas p30 is found in the fraction enriched with sperm heads. This latter calmodulin binding protein, p30, appears to be associated with the perinuclear theca. None of these binding proteins was solubilized by nonionic detergents. Sodium dodecyl sulfate was effective solubilizing p32, whereas p30 was extracted only in conditions reported to isolate the perinuclear theca. Cellular localization of calmodulin binding proteins was also achieved by incubating spermatozoa fixed on slides with biotinylated calmodulin and revealed in a further step by fluorescein-conjugated streptavidin. Using this procedure, it was found that calmodulin binds to the sub and postacrosomal areas of the sperm head along with the midpiece in the presence of Ca(2+). Only a sharp band of fluorescence at the subacrosomal area was observed when this procedure was performed in the absence of Ca(2+) in the presence of EGTA. The pattern of cellular calmodulin binding was highly decreased when spermatozoa were incubated under capacitating conditions, in the presence of heparin, in agreement with the published effect of capacitation on calmodulin binding proteins. PMID- 10819796 TI - Inhibition of rat testicular androgenesis by a polychlorinated biphenyl mixture aroclor 1248. AB - Polychlorinated biphenyls (PCBs) are complex mixtures of congeners that exhibit carcinogenic and toxicant activities in a variety of mammalian tissues. Here, we studied the acute in vivo and in vitro effects of a commercially used PCB product, Aroclor 1248 (A1248), a mixture of tri-, tetra-, and pentachloro congeners. Single intraperitoneal (i.p.) or bilateral intratesticular (i.t.) injections of A1248 decreased serum androgen levels in both groups 24 h after injection. Chorionic gonadotropin-stimulated androgen production by acute testicular cultures from both groups was also reduced, and progesterone production was attenuated in cultures from i.t.-treated animals. The capacity of the postmitochondrial fractions from testes of i.t.-treated animals to convert pregnenolone to progesterone and progesterone to testosterone was reduced as well. In vitro studies revealed that a 10- to 15-min exposure of postmitochondrial testicular fractions and intact interstitial cells from normal animals to A1248 in a subnanomolar concentration range was sufficient to attenuate the conversion of pregnenolone to progesterone and progesterone to testosterone. At micromolar concentrations, A1248 added in vitro also inhibited the conversion of Delta(4)-androstendione to testosterone without affecting the viability of interstitial cells. These results indicate that A1248 down-regulates the testicular androgenesis by an acute inhibition of 3beta-hydroxysteroid dehydrogenase, 17alpha-hydroxylase/lyase, and 17beta-hydroxysteroid dehydrogenase activities. PMID- 10819797 TI - Incidence of cardiac dysrhythmias in patients during pulmonary artery catheter removal after cardiac surgery. AB - OBJECTIVE: No published studies focus on determining the frequency of dysrhythmias during pulmonary artery catheter removal by use of a standard technique. The objective was to assess the incidence and hemodynamic effect of dysrhythmias in patients who had recently undergone cardiac surgery (within 24 hours of when dysrhythmia was noted). METHODS: The study used an electrocardiogram recording by continuous rhythm strip during removal of the pulmonary artery catheter. Pulmonary artery catheters were removed by use of a standard procedure. Data were collected with respect to patients' serum potassium levels, acid-base status, and if they were receiving intravenous antiarrhythmic or inotropic drugs during the PAC removal procedure. RESULTS: One hundred pulmonary artery catheters were removed, with the following types of dysrhythmia recorded: 2 patients had self-terminating ventricular tachycardia with transient hypotension; 1 patient had supraventricular tachycardia without hypotension; 13 patients had isolated ventricular ectopic dysrhythmia without hypotension; 2 patients had isolated atrial ectopic dysrhythmia without hypotension; and 1 patient had isolated atrial ectopic dysrhythmia and 3-beat supraventricular tachycardia without hypotension. CONCLUSION: The use of a standard technique for pulmonary artery catheter removal demonstrated a 2% incidence of nonsustained ventricular tachycardia associated with transient hypotension. Fewer incidences of dysrhythmias were noted in the patients (4 of 29) who had abnormal serum potassium levels, abnormal pH, or pharmacologic association during catheter removal in comparison with those patients without this association (15 patients of 71). No statistically significant difference was noted in the incidence of dysrhythmia during pulmonary artery catheter removal between these 2 groups (chi(2) = 0.72, P =.39). PMID- 10819798 TI - Perceived learning needs of the patient undergoing coronary angioplasty: an integrative review of the literature. AB - OBJECTIVE: This study presents, through an integrative review, a comprehensive account of the perceived concerns and learning needs of patients in the early recovery period after a coronary angioplasty. SCOPE: Nineteen studies involving the patient who has undergone coronary angioplasty were identified using CINAHL and MEDLINE. These studies were examined to compare samples, methods, findings, implications, and suggestions for future research. FINDINGS: Overall the subjects believed that undergoing coronary angioplasty was positive and beneficial, and they viewed it as a minimally invasive, routine procedure. Informational knowledge, such as risk factor education and survival management, were considered of high importance. The majority of subjects modified their behavior, and the most common modification was in diet. Both learned knowledge and lifestyle changes decreased over time. Self-efficacy expectations and levels of anxiety were predictors of behavior changes and knowledge retention in the early recovery period after the coronary angioplasty procedure. IMPLICATIONS: Health professionals must emphasize the seriousness and long-term outcomes of untreated heart disease. Education programs should be individualized and streamlined. Spouses and significant others have informational needs and should be included in education programs. Learning needs in the acute care setting differ from those in the outpatient setting. Continuing education and resources need to be available for patients who are recovering from percutaneous transluminal coronary angioplasty and their families, and should build upon knowledge obtained during hospitalization. PMID- 10819799 TI - A pilot study exploring mood state and dyspnea in mechanically ventilated patients. AB - OBJECTIVE: The objective of this study was to explore preweaning mood state and dyspnea in mechanically ventilated patients. METHODS: Before ventilatory weaning, 21 critically ill patients completed the short profile of mood states (higher scores equal greater disturbance), and a 10 cm dyspnea visual analogue scale (none to extremely severe). Weaning outcome at 24 hours was recorded. RESULTS: The mean +/- SD total mood disturbance (possible range, 0 to 16) and subscale scores (possible range, 0 to 4) were as follows: total, 6.10 +/- 4.06; tension, 1.07 +/- 0.64; depression, 1.16 +/- 0. 93; anger, 1.05 +/- 0.82; vigor, 1.04 +/- 0.84; fatigue, 1.96 +/- 0. 90; and confusion, 1.27 +/- 0.91. Mean dyspnea was 3.22 +/- 2.26 cm. Dyspnea intensity correlated negatively with vigor (r = -0.38, P <. 10). Lower preweaning vigor tended to differentiate successful from unsuccessful weaning (Mann-Whitney U = 22.0; P =.07). CONCLUSION: Patients who weaned successfully experienced greater mood disturbance. Moderate mood disturbance may be a necessary stimulus for successful weaning. PMID- 10819800 TI - Research on the quality of life of lung transplant candidates and recipients: an integrative review. AB - Although lung transplantation is one of the most rapidly growing areas of solid organ transplantation, there has been little research on the quality of life of lung transplant candidates or recipients. This review critiques and synthesizes the quality-of-life reports concerning these patients that have been published between January 1980 and January 1999. The purposes of this review of the literature were to (1) examine both the conceptual and operational definitions of quality of life used; (2) identify and list the instruments used to measure quality of life; (3) investigate methodologic issues; and (4) determine the state of-the-art of research in this area. On the basis of this review, suggestions are made for future studies. PMID- 10819801 TI - Family assessment tools: a review of the literature from 1978-1997. AB - Traditionally, nursing practice in critical care settings has been focused on recognizing and addressing the needs of the patient with an acute and serious health problem and individual family members. Little progress has been made in understanding how families manage this hospitalization experience; however, family health has been reported to be a significant factor in the patient's recovery. The purpose of this article is to review the literature from 1978 to 1997 that has examined family assessment tools in a variety of clinical settings. The ultimate goal of the review is to determine their usefulness for critical care environments and their congruence with family systems nursing, which is aimed at the cognitive, behavioral, and affective domains of family functioning. The following characteristics are used to review each of the selected instruments: theoretical framework; purpose; description; the unit of analysis; ease of administration and scoring; reading level; psychometric evaluation; and utility to guide clinical practice and research. Although the instruments have a variety of strengths, none of them are congruent with the philosophy of family systems nursing. Therefore instruments need to be developed that would guide assessment and interventions for nurses in critical care settings. PMID- 10819802 TI - Advanced practice nursing role delineation in acute and critical care: application of the strong model of advanced practice. AB - PURPOSE: This purpose of this study was to differentiate between the roles of clinical nurse specialists and acute care nurse practitioners. BACKGROUND AND SIGNIFICANCE: Hypothesized blending of the clinical nurse specialist and acute care nurse practitioner roles is thought to result in an acute care clinician who integrates the clinical skills of the nurse practitioner with the systems knowledge, educational commitment, and leadership ability of the clinical nurse specialist. Ideally, this role blending would facilitate excellence in both direct and indirect patient care. The Strong Model of Advanced Practice, which incorporates practice domains of direct comprehensive care, support of systems, education, research, and publication and professional leadership, was tested to search for practical evidence of role blending. METHODS: This descriptive, exploratory, pilot study included subjects (N = 18) solicited from an academic medical center and from an Internet advanced practice listserv. Questionnaires included self-ranking of expertise in practice domains, as well as valuing of role-related tasks. Content validity was judged by an expert panel of advanced practice nurses. RESULTS: Analyses of descriptive statistics revealed that clinical nurse specialists, who had more experience both as registered nurses and in the advanced practice nurse role, self-ranked their expertise higher in all practice domains. Acute care nurse practitioners placed higher importance on tasks related to direct comprehensive care, including conducting histories and physicals, diagnosing, and performing diagnostic procedures, whereas clinical nurse specialists assigned greater importance to tasks related to education, research, and leadership. CONCLUSIONS: Levels of self-assessed clinical expertise as well as valuing of role-related tasks differed among this sample of clinical nurse specialists and acute care nurse practitioners. Groundwork has been laid for continuing exploration into differentiation in advanced practice nursing roles. IMPLICATIONS: As the clinical nurse specialist role changes and the acute care nurse practitioner role emerges, it is imperative that advanced practice nurses describe their contribution to health care. Associating advanced practice nursing activities with outcomes will help further characterize these 2 advanced practice roles. PMID- 10819803 TI - Acinetobacter baumanii line-associated infection. PMID- 10819804 TI - Asthma education: creating a partnership. AB - This article advances the theory that the key to creating an effective partnership is teaching asthma patients what to self-treat, how to self-treat, and when to consult a clinician. The five comanaging rules that the health educator is encouraged to emphasize with the adult asthma patient are: know your own unique asthma symptoms and triggers; keep written records; see appropriate specialists; know your medicines and follow your action plan; and accept no treatment you do not understand. Current research shows asthma to be a chronic inflammatory disorder of the airways. In susceptible individuals, this inflammation causes recurrent episodes of wheezing, breathlessness, chest tightness, and cough, particularly at night and in the early morning. The stepwise approach to asthma therapy divides asthma into several levels of severity. However, patients at any level of severity can have mild, moderate, or severe exacerbations. Asthma triggers; how to use a metered dose inhaler (MDI), a dry powder inhaler (DPI), and a peak flow meter; and how to follow an asthma action plan are thoroughly covered. The last section of the article deals at length with the indications for and actions of long-term-control medications, used to achieve and maintain control of persistent asthma, and quick-relief medications, used to treat symptoms and exacerbations. PMID- 10819806 TI - Prospective trial of early feeding and bowel stimulation after radical hysterectomy. AB - OBJECTIVE: In an attempt to decrease hospital stay we performed a prospective trial of aggressive bowel stimulation and early postoperative feeding after radical hysterectomy. STUDY DESIGN: In a prospective trial of 20 consecutive patients undergoing class 3 radical hysterectomy, feeding of a clear liquid diet and bowel stimulation with oral 66% sodium phosphate solution (Fleet Phospho Soda) were instituted on postoperative day 1. Patients were discharged after passage of flatus or stool. RESULTS: Median time to discharge was 3.5 days. No patient had emesis, ileus, or bowel obstruction. The decrease in hospital stay with respect to those in our previous trial with traditional postoperative feeding and our original study on postoperative bowel stimulation was statistically significant. CONCLUSION: Aggressive bowel stimulation with Fleet Phospho-Soda and early feeding after radical hysterectomy resulted in early return of bowel function and early discharge without significant intestinal complication. PMID- 10819807 TI - Gap junctional intercellular communication and connexin 43 expression in ovarian carcinoma. AB - OBJECTIVES: Gap junctions, which are composed of subunits termed connexins, are plasma membrane channels that link the interior of adjacent cells and permit cells to directly exchange small molecules and ions. Loss or dysfunction of gap junctions appears to be important in allowing cancer cells to escape growth regulation. In a previous study we showed that human ovarian surface epithelial cells exhibited extensive gap junctions and expression of connexin 43. These were nearly absent in human ovarian adenocarcinoma cell lines. To ensure that this variation was not artificially produced by culturing techniques, this study evaluated gap junctions and connexin 43 expressions in normal ovaries and in ovarian adenocarcinomas. STUDY DESIGN: Specimens of normal ovaries and ovarian adenocarcinomas were obtained at the time of surgery and flash-frozen in liquid nitrogen. Connexin 43 immunostaining was performed on all specimens. RESULTS: Among the 11 normal ovaries an average of 59% of the surface epithelium stained positively for connexin 43. In contrast, among the 10 ovarian adenocarcinomas only 19% of each specimen stained positively for connexin 43 (P =.01). CONCLUSION: Similar to our studies on human ovarian surface epithelial cells and ovarian adenocarcinoma cell lines, surgical specimens of normal ovary exhibited extensive connexin 43 expression, whereas connexin 43 expression was nearly absent in ovarian adenocarcinomas. It thus appears that the previously reported loss of gap junctions and connexin 43 was actually associated with a neoplastic process, rather than being artificially induced in the laboratory. PMID- 10819808 TI - Predictors of success after embryo transfer: experience from a single provider. AB - OBJECTIVE: Our goal was to examine the variables present at the time of embryo transfer and to determine their effects on the clinical pregnancy rate. STUDY DESIGN: All fresh and frozen embryo transfers during a 3-year period in a university-based in vitro fertilization program were examined. Female age, previous in vitro fertilization attempt, diagnosis, embryo number and quality, transfer technique, and presence of a clinical pregnancy were recorded for each couple. Logistic regression analyses were performed both univariately and multivariately to determine the association between a clinical pregnancy and the independent variables. RESULTS: All transfers during the study period were included in the analysis. The four primary diagnoses were pelvic or tubal disease, male factor infertility, unexplained infertility, and endometriosis. The 46 frozen embryo transfers had a clinical pregnancy rate similar to that among the 159 fresh embryo transfers and were therefore included in the analysis. One variable was found to significantly affect the outcome, the number of high-grade embryos placed. The presence of a previous failed embryo transfer tended to lower the success rate for future attempts; however, this result did not reach statistical significance. The catheter type and the transfer difficulty did not affect outcome. CONCLUSION: The two most important variables for predicting a clinical pregnancy are a first-time transfer and the number of high-grade embryos placed. Neither the type of embryo transfer catheter used nor the diagnosis affected outcome. In this small sample difficult embryo transfers did not diminish the chance for a successful outcome. PMID- 10819809 TI - Prediction of endometrial ablation success according to perioperative findings. AB - OBJECTIVE: The aim of this study was to determine which factors in the perioperative period influence the success of endometrial ablation in alleviating menorrhagia. STUDY DESIGN: We performed a retrospective chart review of 120 women aged 27 to 49 years who underwent endometrial ablation after 2 months of preoperative treatment with danazol (Danocrine, 800 mg/d orally) or leuprolide (Lupron, 3.75 mg in one intramuscular injection each month). Patients who required medical management or additional operations to control the vaginal bleeding during follow-up (median follow-up, 37 weeks) were considered to have ablation failures. RESULTS: Sixty-three percent of patients (76/120) had a successful procedure. The chance of success was greater if a cavity of normal appearance was found (odds ratio, 2.3; P =.04). The finding of an intramural fibroid before the procedure resulted in a reduced trend toward success (odds ratio, 0.4; P =.06). The use of danazol pretreatment improved the rate of success overall (odds ratio, 2.2; P =.05) and especially among women <40 years old (P =.01) CONCLUSION: Perioperative findings may provide useful information in counseling patients regarding endometrial ablation. Success is greater among patients with a normal intrauterine cavity and after preoperative treatment with danazol. PMID- 10819811 TI - Presence of ribonucleic acid in human spermatozoa: differences in content between normal and abnormal spermatozoa. AB - OBJECTIVE: Our purpose was to determine whether there are any differences in total ribonucleic acid content between normal and abnormal human spermatozoa. STUDY DESIGN: Spermatozoa were obtained from men undergoing routine semen analysis at a university-based reproductive genetics laboratory. Specimens were classified as normal or abnormal according to World Health Organization criteria. Total ribonucleic acid was removed by acid-phenol extraction, and ribonucleic acid expression levels were determined by spectrophotometric analysis. RESULTS: Abnormal spermatozoa were found to have significantly more ribonucleic acid (0.14 +/- 0.02 mg/10(6) spermatozoa) than normal spermatozoa (0.05 +/- 0.01 mg/10(6) spermatozoa; P <.001). CONCLUSION: Ribonucleic acid content is significantly altered in abnormal spermatozoa, and this alteration may be the result of some defect in the posttranscriptional pathway. PMID- 10819810 TI - A randomized trial of intravaginal nonoxynol 9 versus oral metronidazole in the treatment of vaginal trichomoniasis. AB - OBJECTIVE: This study was undertaken to investigate the efficacy of nonoxynol 9 suppositories in the treatment of vaginal trichomoniasis. STUDY DESIGN: In this prospective comparison trial 46 women with documented motile trichomonads found on a wet preparation were randomly assigned to one of two treatment arms: (1) a single oral dose of 2 g metronidazole and (2) a single 150-mg nonoxynol 9 suppository placed intravaginally for 3 consecutive nights. Cure was determined by a repeated wet preparation examination. After its first year, the study was terminated because of the poor efficacy of the nonoxynol 9 suppositories. RESULTS: Results were available for 33 patients. Three of 17 patients treated with nonoxynol 9 had negative wet preparation results at retest (17.6% cure rate). All 16 patients treated with metronidazole had negative wet preparation results (100% cure rate). All women with nonoxynol 9 failures who were evaluated after treatment with 2 g metronidazole had negative wet preparation results. CONCLUSION: Intravaginal nonoxynol 9 at the tested dose and by the tested method of delivery was not an effective cure for vaginal trichomoniasis. PMID- 10819812 TI - Type 5 phosphodiesterase regulation of human sperm motility. AB - OBJECTIVE: Inhibition of phosphodiesterases results in the buildup of intracellular cyclic nucleotides, which have been shown to affect sperm motility and acrosome reaction. The objective of this study was to determine whether the cyclic guanosine monophosphate-specific type 5 phosphodiesterase inhibitor sildenafil has an effect on sperm motility and acrosome parameters. STUDY DESIGN: Sperm cells were washed by two-layer colloid wash and resuspended in modified human tubal fluid with 5% serum albumin. They were incubated in the presence of different concentrations (0-40 nmol/L) of the type 5 phosphodiesterase inhibitor sildenafil. Aliquots of sperm were removed at hours 0, 4, 24, and 48, and motility parameters were measured on the Hamilton-Thorn HTM-C (Hamilton-Thorn Research, Danvers, Mass) motility analyzer. Sperm acrosomes were analyzed with the Spermac (Stain Enterprises, South Africa; distributed by Sage Biopharma, Bedminster, NJ) acrosome stain. RESULTS: Sperm progressive motility and hyperactivation were stimulated to greater than the control at hour 4, followed by a decrease. There was a dose-dependent effect of the type 5 phosphodiesterase inhibitor on sperm motility parameters but not on percentage of cells with acrosome reaction. The type 5 phosphodiesterase inhibitor stimulated sperm acrosome reaction by almost 50% above the control. CONCLUSION: These results suggest that inhibition of type 5 phosphodiesterase activity in human sperm resulted in enhanced progressive motility and hyperactivation. In addition, inhibition of type 5 phosphodiesterase also caused an increase in acrosome reaction. This suggests a role for type 5 phosphodiesterase in preventing premature acrosome reaction, which is associated with failed fertilization. PMID- 10819813 TI - Analysis of papillomavirus consensus L1 gene in a closed colony of baboons (Papio anubis). AB - OBJECTIVE: The purpose of this study was to analyze cervical specimens and semen from a closed colony of baboons for the presence of the papillomavirus consensus L1 gene. STUDY DESIGN: Cervical swabs were collected from lightly anesthetized female baboons. Semen was collected from a male baboon by standard electroejaculation techniques. Deoxyribonucleic acid was extracted from the cells by two different methods and analyzed by polymerase chain reaction targeting the L1 consensus gene common for >25 genital papillomaviruses. RESULTS: Analyses of the polymerase chain reaction-amplified products did not reveal bands for the papillomavirus in either the cervical specimens or the semen. CONCLUSIONS: The hypothesis of a linkage between primates with papillomavirus as a common factor is not supported by the results of this study. This information is also important in assisting clinicians in setting up specific pathogen-free colonies of baboons. PMID- 10819814 TI - Changing trends in patient decisions concerning genetic amniocentesis. AB - OBJECTIVE: This study was undertaken to determine whether there was a change in patient decisions concerning genetic amniocentesis during the period 1995-1998. STUDY DESIGN: All patients referred for genetic counseling because of advanced maternal age, abnormal serum triple-screen results, or ultrasonographic abnormalities between January and March 1995 and between January and March 1998 were evaluated through a retrospective chart review. Patient characteristics included age, race, and gestational age. Group 1 consisted of patients from 1995. Group 2 consisted of patients from 1998. Data on patient decisions concerning amniocentesis before and after genetic counseling and ultrasonographic examination were compared in each group. Groups 1 and 2 were then compared with respect to decisions before and after genetic counseling and ultrasonographic evaluation. RESULTS: A total of 112 patients were studied. Group 1 consisted of 53 patients and group 2 consisted of 59 patients. When the groups were compared, no differences in age, race, or gestational age were noted. In group 1, before counseling, 18 of 53 patients desired genetic testing, compared with 44 of 53 after counseling (P =.02). In group 2, before counseling, 4 of 59 patients desired genetic testing, compared with 15 of 59 after counseling (P =.01). A significantly greater number of patients in group 1 than in group 2 desired genetic testing both before counseling (n = 18/53 vs n = 4/59; P =.01) and after counseling (n = 44/53 vs n = 15/59; P =.01). CONCLUSION: Fewer patients at risk for Down syndrome in 1998 than in 1995 desired amniocentesis both before and after genetic counseling and ultrasonographic examination. PMID- 10819815 TI - Urinary incontinence and pelvic organ prolapse in women with Marfan or Ehlers Danlos syndrome. AB - OBJECTIVE: This study was undertaken to determine the prevalence of urinary incontinence and pelvic organ prolapse in a cohort of women with either Marfan syndrome or Ehlers-Danlos syndrome. STUDY DESIGN: Female patients with either Marfan syndrome or Ehlers-Danlos syndrome were identified through a medical records search at two urban hospitals. Each patient's medical record was reviewed, and the history of pelvic organ prolapse and urinary incontinence was obtained through telephone interview. RESULTS: Twelve women with Marfan syndrome were identified. Among these women 5 (42%) reported a history of urinary incontinence and 4 (33%) reported a history of pelvic organ prolapse. Eight women with Ehlers-Danlos syndrome were identified. Among these women 4 (50%) reported a history of urinary incontinence and 6 (75%) reported a history of pelvic organ prolapse. CONCLUSIONS: Women with Marfan or Ehlers-Danlos syndrome have high rates of urinary incontinence and pelvic organ prolapse. This finding supports the hypothesized etiologic role of connective tissue disorders as a factor in the pathogenesis of these conditions. PMID- 10819816 TI - Gestational diabetes screening in subsequent pregnancies of previously healthy patients. AB - OBJECTIVE: Our purpose was to evaluate women without gestational diabetes mellitus in an index pregnancy for the likelihood that gestational diabetes would develop and for risk factors for carbohydrate intolerance in a subsequent pregnancy. STUDY DESIGN: A retrospective review of medical records at a teaching hospital universally screening for gestational diabetes identified multiparous women who had been delivered twice between 1994 and 1997 and who, in the first (index) pregnancy, had had a normal result on a screening test with 50 g of glucose used in a "glucola" beverage (< or =140 mg/dL). RESULTS: In this population with normal glucose screening values in the index pregnancy, 352 (92.4%) of 381 women had at least one risk factor for gestational diabetes. However, none of the 381 women had gestational diabetes in the subsequent pregnancy (0/381, 95% confidence interval < or =1%), including 45 (12. 4%) who had an abnormal result on the 50-g glucose screening test. Regression analysis showed this test result in the index pregnancy (P =.001) to be the only studied variable significantly associated with the 50-g glucose value in the subsequent pregnancy. CONCLUSION: Despite a high rate of risk factors for gestational diabetes, women in our population with a normal glucose value in an index pregnancy have a minimal risk (<1%) that gestational diabetes will develop in a subsequent singleton pregnancy within 4 years. This factor may be included in determining whether women should undergo screening for gestational diabetes. PMID- 10819817 TI - Appendicitis in pregnancy: new information that contradicts long-held clinical beliefs. AB - OBJECTIVE: Our purpose was to elicit a better understanding of the presentation of acute appendicitis in pregnancy and to clarify diagnostic dilemmas reported in the literature. STUDY DESIGN: We retrospectively reviewed 66,993 consecutive deliveries from 1986 to 1995 by a computer program. Selected records were reviewed for gestational age; signs and symptoms at presentation; complications including preterm contractions, preterm labor, and appendiceal rupture; and histologic diagnosis of appendicitis. RESULTS: Of 66, 993 deliveries, 67 (0.1%) were complicated by a preoperative diagnosis of probable appendicitis. Acute appendicitis was confirmed histologically in 45 (67%) of the 67 cases, for an incidence of 1 in 1493 pregnancies in this population. Distribution of suspected appendicitis in pregnancy was as follows: first trimester, 17 cases (25%); second trimester, 27 (40%); and third trimester, 23 (34%). Right-lower-quadrant pain was the most common presenting symptom regardless of gestational age (first trimester, 12 [86%] of 14 cases; second trimester, 15 [83%] of 18 cases; and third trimester, 10 [78%] of 13 cases). The mean maximal temperature for proven appendicitis was 37.6 degrees C (35.5 degrees C-39.4 degrees C), in comparison with 37.8 degrees C (36.7 degrees C-38.9 degrees C; not significant) for those with normal histologic findings. The mean leukocyte count in patients with proven appendicitis was 16.4 x 10(9)/L (8.2-27.0 x 10(9)/L), in comparison with 14.0 x 10(9)/L (5. 9-25.0 x 10(9)/L) for patients with normal histologic findings. At the time of surgery, perforation had occurred in 8 cases. Of 23 patients at > or =24 weeks' gestational age, 19 (83%) had contractions and an additional 3 patients (13%) had preterm labor with documented cervical change. One patient was delivered in the immediate postoperative period because of abruptio placentae. CONCLUSION: Pain in the right lower quadrant of the abdomen is the most common presenting symptom of appendicitis in pregnancy regardless of gestational age. Fever and leukocytosis are not clear indicators of appendicitis in pregnancy and preterm labor is a problem after appendectomy, but preterm delivery is rare. PMID- 10819818 TI - Transvaginal ultrasonographic cervical measurement as a predictor of successful labor induction. AB - OBJECTIVE: Our purpose was to compare transvaginal cervical measurement and the Bishop score as indicators of duration of labor and successful induction of labor at term. STUDY DESIGN: This prospective observational study recruited women with singleton gestations scheduled for induction of labor at > or =37 weeks. Transvaginal ultrasonographic measurement of cervical length was performed and the Bishop score was determined, each by operators masked to the other measurement. Data were collected on parity, gestational age, mode of delivery, induction agent, induction-to-delivery interval, Bishop score, and cervical length measurement. RESULTS: A total of 77 women were analyzed. Vaginal delivery occurred in 69%. Both Bishop score and cervical length showed linear correlation with duration of labor (R(2) = 0.43, P <. 001; R(2) = 0.48, P <.001; respectively). Women with cervical length <3.0 cm had shorter labors (P <.001) and were more likely to be delivered vaginally (P <.001). Women with a Bishop score >4 also had shorter labors and were more likely to be delivered vaginally, with similar P values. A logistic regression model identified cervical length and parity as the only independent predictors of vaginal delivery. CONCLUSIONS: Both ultrasonographically measured cervical length and Bishop score predict duration of labor and likelihood of vaginal delivery. However, only cervical length and parity were independent predictors of mode of delivery. PMID- 10819819 TI - Postoperative morbidity in the morbidly obese parturient woman: supraumbilical and low transverse abdominal approaches. AB - OBJECTIVE: Our purpose was to determine the differences in postoperative morbidity in obese women who had a supraumbilical or a Pfannenstiel incision at cesarean delivery. STUDY DESIGN: A case-control retrospective review was conducted of all patients who were at >150% ideal body weight when undergoing cesarean delivery between 1989 and 1995 by means of either a supraumbilical or a Pfannenstiel incision. Patients were excluded if medical records were unavailable. A total of 15 women who had a supraumbilical incision and 54 who had a low transverse incision were included in the analysis. Antenatal complications were examined, as were age, weight, and training level of the surgeon. Postoperative complications were then compared. RESULTS: The groups were similar in age and antepartum complications. However, mean weight and percentage of ideal body weight in the supraumbilical group were both higher (P <.00001 and P <.0001, respectively), with the supraumbilical group 83 lb heavier on average. No significant differences were seen in any postoperative complication. CONCLUSION: Postoperative morbidity in morbidly obese women undergoing cesarean delivery does not differ between a supraumbilical approach and the low transverse abdominal incision. PMID- 10819820 TI - Demographics, management, and outcome of peripartum cardiomyopathy in a community hospital. AB - OBJECTIVE: The purpose of this study was to describe the outcome of peripartum cardiomyopathy in patients cared for in a community hospital. STUDY DESIGN: The cases of peripartum cardiomyopathy treated at Central Baptist Hospital in Lexington, Kentucky, from January 1, 1992, to December 31, 1998, were reviewed. RESULTS: Eleven patients with peripartum cardiomyopathy were identified. The patient population was 91% white and 9% African American. Seventy-two percent of patients were nulliparous, and the prevalence of chronic hypertension was 27%. All patients were examined with echocardiography and met diagnostic criteria for the disease when this modality was used. The mean ejection fraction was 32% +/- 10%. Invasive techniques used to assist in diagnosis included left ventricular catheterization (63%), right ventricular catheterization (54%), and cardiac biopsy (54%). One patient required cardiac transplantation. This patient also had an embolic stroke from a confirmed mural thrombus. No study patient died of the disease, and no other major complications were observed. CONCLUSIONS: The patient profile of peripartum cardiomyopathy in this study differed remarkably from profiles in published reports. Nulliparous white women have better outcomes than indicated by previous reports, probably because of the low frequency of coexisting chronic disease and a younger age at diagnosis. PMID- 10819821 TI - A randomized trial of misoprostol versus extra-amniotic sodium chloride infusion with oxytocin for induction of labor. AB - OBJECTIVE: Our purpose was to compare the efficacy and safety of misoprostol and extra-amniotic sodium chloride infusion with oxytocin for induction of labor. STUDY DESIGN: This randomized trial compared two methods of labor induction in women requiring cervical ripening. One hundred twenty-three women undergoing labor induction with a Bishop score < or =5 were randomly selected to receive either misoprostol, 50 microg intravaginally every 4 hours, or extra-amniotic sodium chloride infusion. The primary outcome variable was the time interval from induction to vaginal delivery. RESULTS: Sixty-one women received extra-amniotic sodium chloride infusion and 62 women received misoprostol. The mean time interval from the start of induction to vaginal delivery was 15.0 +/- 5.0 hours and 16.5 +/- 7.2 hours for the extra-amniotic infusion and misoprostol groups, respectively (P, not significant). The cesarean delivery rate was not significantly different between the 2 groups (32.8% for the extra-amniotic infusion group; 19.4% for the misoprostol group). Maternal and neonatal outcomes were similar between the 2 groups. CONCLUSIONS: Both methods of induction are equally efficacious and result in similar maternal and neonatal outcomes. PMID- 10819822 TI - Semen pH in patients with normal versus abnormal sperm characteristics. AB - OBJECTIVE: The World Health Organization laboratory manual, last revised in 1992, states that the normal pH of semen ranges from 7.2 to 8.0. Our experience has been that values in our patient population are consistently higher than this range. To confirm this we reviewed >1100 semen records. STUDY DESIGN: All patient records from January 1994 to December 1998 that had semen pH measurements and sperm concentration and motility measurements recorded were included in this study. We also determined the semen pH in a subgroup of patients who underwent sperm preparations for intrauterine inseminations that resulted in documented pregnancies. Histograms were used to describe the populations and the Mann Whitney test was used for group comparisons. RESULTS: For all patients (N = 1199) mean (+/-SD) semen pH was 8.2 +/- 0.3. The range was 7.3 to 9.5, with pH <8.0 in 32% of the samples. The semen pH among the patients with normal sperm concentration and motility values (n = 602) was not different from that among those with abnormal parameters (n = 597). Mean semen pH value was 8.2 for both groups. In a small group of patients (n = 19) whose sperm preparations had been documented to result in a clinical pregnancy after intrauterine insemination the semen pH was 8.3 +/- 0.3, with a range of 7.9 to 8.7. CONCLUSION: Our study questions the reference range defined by the World Health Organization for semen pH of 7.2 to 8.0. The mean values that we observed in our population, including those of samples from patients with normal sperm parameters, consistently lay outside that range. PMID- 10819823 TI - The papanicolaou smear: inadequate screening test for bacterial vaginosis during pregnancy. AB - OBJECTIVE: Our purpose was to evaluate the ability of the Papanicolaou smear to identify bacterial vaginosis in comparison with the Amsel clinical criteria. STUDY DESIGN: We retrospectively identified 159 pregnant women screened for bacterial vaginosis with the Amsel criteria who had a contemporaneous Papanicolaou smear and negative results on screening for Chlamydia trachomatis and Neisseria gonorrhoeae. Bacterial vaginosis was identified in 45 women. We used the McNemar chi(2) test to determine discrepancies between the two screening methods for the detection of bacterial vaginosis. RESULTS: Compared with the Amsel criteria, the sensitivity and specificity of the Papanicolaou smear for yielding a diagnosis of bacterial vaginosis were 49% (95% confidence interval, 36%-64%) and 93% (95% confidence interval, 86%-97%), respectively, with a positive predictive value of 73% and a negative predictive value of 82%. The detection of bacterial vaginosis by Papanicolaou smear was significantly different from that by Amsel criteria (P =. 01). CONCLUSION: The Papanicolaou smear is not a reliable screening test for bacterial vaginosis during pregnancy. PMID- 10819824 TI - The effect of obstetric resident gender on forceps delivery rate. AB - OBJECTIVE: This study was undertaken to examine the effect of obstetric resident physician gender on the forceps delivery rate. STUDY DESIGN: Analysis was based on >350,000 deliveries performed by >800 residents in obstetrics and gynecology throughout the United States from 1994 to 1998. A chi(2) analysis was performed on resident statistics from residency review committee report forms. RESULTS: The percentage of total deliveries performed with forceps during residency was significantly higher among male residents (P <. 0001), as was the percentage of vaginal deliveries performed with forceps during residency (P <.0001). The percentage of overall operative vaginal deliveries (vacuum plus forceps) was significantly higher for male residents (P <.0001); however, the percentage of vacuum deliveries did not vary according to gender of the resident when considered independently. CONCLUSION: These results strongly suggest that resident gender affects performance of forceps delivery. PMID- 10819825 TI - Oral glucose tolerance test and the preparatory diet. AB - OBJECTIVE: A 3-day diet containing at least 150 g carbohydrate per day has been used in many centers in preparation for the oral glucose tolerance test. The preparatory diet is thought to reduce false-positive diagnoses of gestational diabetes. The purpose of this study was to evaluate the necessity of a 3-day preparatory diet containing > or =150 g carbohydrate in otherwise healthy pregnant patients. STUDY DESIGN: Twenty healthy obstetric patients with abnormal results on a 1-hour glucose challenge test (> or =140 mg/dL) were enrolled in this prospective pilot study. Two oral glucose tolerance tests were administered. The first was with no dietary restrictions, and the second test was performed after a 3-day diet containing at least 150 g carbohydrate. Patients were given a 3-day dietary supplement, which contained 150 g carbohydrate per day. A food diary verified compliance with the diet and indicated other food intake. RESULTS: There was no difference in the number of oral glucose tolerance tests with abnormal results, with or without the diet (5 in each group). Additionally, no significant difference was found in the mean glucose values in the diet versus no diet groups. CONCLUSION: A preparatory diet does not significantly alter the results of an oral glucose tolerance test administered to healthy pregnant women. The diet unnecessarily delays the diagnosis of gestational diabetes. PMID- 10819826 TI - Obstetric conditions and erythropoietin levels. AB - OBJECTIVE: Our purpose was to evaluate and compare erythropoietin levels as related to obstetric conditions, including acute and chronic bleeding, preeclampsia, and multiple gestations. STUDY DESIGN: During April 1999 all women in the labor and delivery unit with delivery expected to occur within 24 to 72 hours of admission had erythropoietin and hematocrit values obtained. First trimester hematocrit values, obstetric problems, medications, and history of vaginal bleeding were obtained from patient interview, examination, and the prenatal record. Statistics were analyzed by the Student t test and chi(2). RESULTS: During a 1-month period, 302 consecutive women were divided into 5 groups on the basis of obstetric events. Group 1 consisted of women with normal, uncomplicated term singleton gestations (n = 230); group 2, women with acute vaginal bleeding (n = 10); group 3, women with chronic vaginal bleeding (n = 29); group 4, women with multiple gestations (n = 13); and group 5, women with preeclampsia (n = 16). The mean erythropoietin level in group 1 (20. 2 +/- 10.3 mU/mL) was significantly different from values in the other 4 groups (group 2, 74.2 +/- 29.2 mU/mL; group 3, 65.0 +/- 33.0 mU/mL; group 4, 34.8 +/- 16.8 mU/mL; group 5, 43.4 +/- 11.4 mU/mL; P <.001). The admission hematocrit for group 1 (0.369 +/- 0.029) was significantly greater than for groups 2 and 3 (group 2, 0.323 +/- 0. 024; group 3, 0.321 +/- 0.023; P <.001) and significantly lower than for group 5 (0.384 +/- 0.022; P <.05). CONCLUSION: The maternal serum erythropoietin level varies depending on the events occurring during gestation. Acute and chronic bleeding, multiple gestations, and preeclampsia are all associated with various serum erythropoietin levels. PMID- 10819827 TI - Is the management of epidural analgesia associated with an increased risk of cesarean delivery? AB - OBJECTIVE: The aim of this study was to quantify the association of cesarean delivery with epidural analgesia management, specifically with the timing of epidural catheter placement in relation to labor, the type of epidural analgesia, and the use of bolus dosing. STUDY DESIGN: A retrospective cohort design was used to investigate 1561 consecutive nulliparous parturients whose labor occurred between November 1, 1996, and June 30, 1997, at Northwestern Memorial Hospital and who were delivered of term, singleton neonates in a cephalic presentation. The relationship between the management of epidural analgesia and the risk for cesarean delivery was determined with stepwise logistic regression to control for potential confounding variables. RESULTS: There was a significantly increased risk of cesarean delivery associated with decrements in cervical effacement (P =.001), cervical dilatation (P =.001), and fetal station (P =.001) at the time of epidural catheter placement. An increasing number of epidural boluses during the first stage of labor was also associated with increased risk of cesarean delivery (P =.001). After we controlled for maternal age, maternal body mass index, gestational age, infant birth weight, induction of labor, use of magnesium sulfate, and presence of chorioamnionitis, the adjusted odds of cesarean delivery associated with fetal station (odds ratio, 1.45; 95% confidence interval, 1.2 1.7) and epidural boluses (odds ratio, 1.55; 95% confidence interval, 1.3-1.8) during the first stage of labor remained significant. CONCLUSION: The management of epidural analgesia during labor was associated with the potential for increased risk of cesarean delivery. This risk increased with higher stations of the fetal head at the time of epidural catheter placement and with more frequent epidural boluses of local anesthetic during the first stage of labor. PMID- 10819829 TI - The effects of physical abuse on pregnancy outcomes in a low-risk obstetric population. AB - OBJECTIVE: This study was undertaken to evaluate the impact of physical abuse on pregnancy outcomes. STUDY DESIGN: This prospective investigation identified all women seen in the outpatient obstetric clinic with a history of physical abuse and matched each with the next 2 women seen without a history of physical abuse (case/control ratio, 1:2). RESULTS: Twenty-eight abused women were matched with 56 control subjects. The frequencies of preterm birth, mode of delivery, Apgar scores <7 at 5 minutes, umbilical artery pH <7.10 at birth, and unhappy or ambivalent feelings about the pregnancy were not statistically different between the 2 groups. Admission to the neonatal intensive care unit was more common among infants of the abused women (n = 4 [14.2%] vs n = 2 [3.6%]), despite heavier birth weights in the abused group (3501 +/- 581 g vs 3200 +/- 549 g; P =. 023), but this trend did not reach significance. CONCLUSION: A history of physical abuse did not result in adverse pregnancy outcomes, although neonatal intensive care unit admission was more likely. PMID- 10819828 TI - Safety of intra-amniotic digoxin administration before late second-trimester abortion by dilation and evacuation. AB - OBJECTIVE: The purpose of this study was to determine the safety of intra amniotic digoxin injection before late second-trimester pregnancy termination by dilation and evacuation through an assessment of maternal systemic digoxin absorption, cardiac rhythm, and coagulation parameters. STUDY DESIGN: Pregnant women at between 19 and 23 weeks' gestation received 1.0 mg digoxin through intra amniotic injection and then had serum digoxin levels determined for 48 hours and Holter cardiac monitoring performed for 24 hours. Clotting parameters were assessed before digoxin injection and 24 hours later, at the time of the dilation and evacuation procedure. RESULTS: Eight patients completed the study. The mean (+/-SD) serum digoxin peak concentration was 0.81 +/- 0.22 microg/L (range, 0.5 1.1 microg/L). The mean (+/-SD) time to peak digoxin concentration was 11.0 +/- 5.55 hours (range, 4-20 hours). Ambulatory cardiac monitoring showed no rhythm or conduction abnormalities associated with digoxin. Prothrombin time, partial thromboplastin time, and fibrinogen levels did not change significantly between determinations before and after the dilation and evacuation procedure (11.5 to 11.4 seconds, 24.1 to 24.4 seconds, and 441 to 475 mg/dL, respectively). CONCLUSION: The maximum digoxin concentration peak achieved after intra-amniotic injection was in the low therapeutic range. No rhythm or conduction abnormalities associated with digoxin were noted by Holter monitoring. Coagulation parameters did not change significantly. On the basis of the limited systemic absorption and the absence of clinically significant cardiac or clotting effects, intra amniotically administered digoxin may be considered safe for use before late second-trimester pregnancy terminations. PMID- 10819830 TI - The relationship of maternal position to the results of brief nonstress tests: a randomized clinical trial. AB - OBJECTIVE: This study was undertaken to determine whether maternal posture (left lateral recumbent vs semi-Fowler position) had any effect on nonstress test results when the test was performed for a shortened period (10 minutes). STUDY DESIGN: In this randomized clinical trial of 108 patients with singleton pregnancies at 32 to 42 weeks' gestation, patients were randomly assigned to a "sitting first" (semi-Fowler position) or a "supine first" (left lateral recumbent position) group at the initial visit. The order of position was alternated at subsequent visits. Ten minutes of fetal heart rate monitoring was performed in each position at each visit. Computer analysis of the nonstress test was used to interpret each 10-minute segment for reactivity. Statistical analyses were performed on the paired nonstress test unit (sitting and supine). RESULTS: There were no adverse clinical outcomes among the participants. Logistic regression analysis showed that both the sequence of the nonstress test and the position were significant and independent factors related to nonstress test reactivity. Tests performed during the second 10 minutes and tests performed with the patient in the semi-Fowler position were more likely to have reactive results. CONCLUSION: The semi-Fowler position is a superior position for conducting a nonstress test in a short period. Use of this position could decrease the need for prolonged monitoring, thus leading to a more time-effective evaluation of patients at risk. PMID- 10819831 TI - Risk factor scoring for predicting venous thromboembolism in obstetric patients. AB - OBJECTIVE: The aim of this study was to develop a risk factor scoring system for the prediction of venous thromboembolism in obstetric patients. STUDY DESIGN: We conducted a retrospective case-control study of all pregnant or postpartum women admitted to the Jack D. Weiler Hospital from 1987 through 1998 with a discharge diagnosis indicating thromboembolism. For each study subject the three women who were delivered immediately before that index patient were selected for the control group. Data collected included the following: history of thrombosis, age, body mass index, previous abdominal surgical procedures, presence of systemic diseases, and blood type. Each patient was assigned a score that was based on the risk factors identified. RESULTS: We identified 21 patients who had sustained thromboembolic events during pregnancy or 6 weeks post partum. Nineteen of the thromboembolic events (90%) were diagnosed during pregnancy, and these cases were distributed throughout gestation-8 (42%) in the first trimester, 2 (10%) in the second trimester, and 9 (48%) in the third trimester. Six (28%) of these patients had pulmonary embolisms. Two cases of postpartum thromboembolic events were documented. Both were cases of pulmonary embolism. There was 1 maternal death. Patients with a score >2 were at significantly increased risk for having a thromboembolism, with an odds ratio of 4.8 (P <.05). The sensitivity, specificity, positive predictive value, and negative predictive value of this cutoff point were 21%, 95%, 57%, and 78%, respectively. CONCLUSION: Obstetric patients with high risk factor scores were at increased risk for thromboembolism. PMID- 10819832 TI - Evaluation of the centrifuged and Gram-stained smear, urinalysis, and reagent strip testing to detect asymptomatic bacteriuria in obstetric patients. AB - OBJECTIVE: Our purpose was to compare the efficacy of the centrifuged and Gram stained smear with the efficacy of both urinalysis and reagent strip testing for nitrites and leukocyte esterase in detecting asymptomatic bacteriuria in obstetric patients. STUDY DESIGN: A midstream urine specimen was evaluated in 528 patients either at the initial prenatal visit or at a visit because of possible preterm labor. Separate aliquots were tested by centrifugation (with a Cytospin Cytocentrifuge; Shandon, Inc, Pittsburgh, Pa) with Gram stain, by microscopic urinalysis for the presence of moderate to large numbers of bacteria or >10 leukocytes per high-power field, and by reagent strips for the presence of nitrites or leukocyte esterase activity. Results were compared with those of a quantitative urine culture obtained with blood and MacConkey agar plates. RESULTS: Thirty-six women (6.8%) had urine cultures showing 100,000 colony forming units of a uropathogen per milliliter. The sensitivity and specificity of testing by centrifugation and Gram stain were 100% and 7.7%, respectively. Urinalysis and dipstick testing offered a sensitivity of 80.6% and 47.2%, respectively, with a specificity of 71.5% and 80.3%. No combination of tests, in series or in parallel, offered improved specificity over urinalysis alone. CONCLUSIONS: Centrifugation with Gram stain of a urine specimen offers excellent sensitivity but very poor specificity compared with microscopic urinalysis for the detection of asymptomatic bacteriuria and is not an acceptable screening test in an obstetric population. The false-negative rates of urinalysis (19.4%) and reagent strip testing (52.8%) preclude these from being excellent screening tests for asymptomatic bacteriuria. Given the potential sequelae of undiagnosed asymptomatic bacteriuria in an obstetric population, we conclude that urine cultures should be used for all pregnant patients to detect asymptomatic bacteriuria. PMID- 10819833 TI - Postpartum depression: a comparison of screening and routine clinical evaluation. AB - OBJECTIVE: This study compared the efficacy of routine clinical evaluation with that of screening with the Edinburgh Postnatal Depression Scale for the detection of postpartum depression in a residency training program practice. STUDY DESIGN: Three hundred ninety-one patients during a period of 1 year were assigned according to delivery date to screening for postpartum depression with the Edinburgh Postnatal Depression Scale or to a control group who had only spontaneous detection during routine clinical evaluation. The incidences of postpartum depression detection and demographic characteristics were compared between 79 patients in the Edinburgh Postnatal Depression Scale group and 96 patients in the clinical evaluation group by means of chi(2) analyses. RESULTS: The incidence of postpartum depression detection with the Edinburgh Postnatal Depression Scale was significantly higher than the incidence of spontaneous detection during routine clinical evaluation (35.4% and 6.3%, respectively; P =.001). CONCLUSIONS: The Edinburgh Postnatal Depression Scale is an effective adjunct to clinical interview for diagnosis of postpartum depression and should be considered in residency training. PMID- 10819835 TI - Laparoscopic removal of a transabdominal cervical cerclage. AB - Cervical incompetence has been acknowledged as a significant entity predisposing patients to second-trimester miscarriage. Various surgical techniques and approaches have been used in an attempt to prolong pregnancy and improve perinatal outcome. These include transvaginal and transabdominal cervical cerclage. Some patients require the placement of a transabdominal cervicoisthmic cerclage. Should the cerclage fail or the patient have preterm premature rupture of membranes, removal of the cerclage may be necessary. As a result the application of laparoscopy for the management of cervicoisthmic cerclage removal has been advocated in an effort to limit surgical complications. We report a case of laparoscopic removal of a transabdominally placed cervical cerclage in a 32 year-old woman at 16 weeks' gestation with preterm premature rupture of membranes and inevitable miscarriage. Laparoscopy appeared to be a safe and effective means of managing the removal of this transabdominally placed cervicoisthmic cerclage. PMID- 10819834 TI - Severe perineal lacerations during vaginal delivery: the University of Miami experience. AB - OBJECTIVE: After childbirth-related third- or fourth-degree perineal lacerations, the estimated incidence of wound disruption, fecal incontinence, or fistula ranges from 1% to 10%. Risk factors associated with severe laceration were analyzed at a single large teaching institution. STUDY DESIGN: This study consisted of an analysis of data from the delivery database of Jackson Memorial Hospital, University of Miami, from 1989 through 1995. Included were vaginal deliveries for which complete information was available on maternal age, parity, ethnicity (white, black, or Hispanic), birth weight, episiotomy versus no episiotomy, type of episiotomy, and delivery (normal spontaneous, vacuum, or forceps). Multiple gestations, cases of shoulder dystocia, cesarean deliveries, patients with a history of cesarean delivery, and babies weighing <500 g at birth were excluded from this study. Both univariate and multivariate analyses were performed with variables such as maternal age, race, birth weight, type of episiotomy if any, and type of vaginal delivery. RESULTS: Among the 71,959 women who were delivered at our institution during the 7-year study period, 50,210 met the inclusion criteria. Through time there had been a decline in the use of episiotomy in general and of midline episiotomy in particular. The annual total number of deliveries also decreased. The episiotomy procedure per se and the type of episiotomy as well as birth weight, assisted vaginal delivery, and older maternal age were identified as independent risk factors associated with third- and fourth-degree perineal lacerations. CONCLUSION: Although episiotomy is an important risk factor for severe lacerations after vaginal delivery, there are other significant independent risk factors, such as maternal age, birth weight, and assisted vaginal delivery, that should be considered in counseling and making decisions regarding delivery modality. Older patients who are being delivered of a first child are at higher risk for severe laceration. Midline episiotomy and assisted vaginal delivery should therefore be avoided in this population whenever possible, especially in the presence of a large baby. PMID- 10819836 TI - The prevalence of thromboembolic events among women with extended bed rest prescribed as part of the treatment for premature labor or preterm premature rupture of membranes. AB - OBJECTIVE: This study was undertaken to determine the prevalence of thromboembolic events among women with extended bed rest prescribed as part of the treatment of premature labor or preterm premature rupture of membranes. STUDY DESIGN: A retrospective chart review was undertaken of all women who had bed rest of >/=3 days' duration prescribed as part of the treatment of premature labor or preterm premature rupture of membranes in the Akron General Medical Center Perinatal Unit during the period January 1, 1997-December 31, 1998. The prevalence of thromboembolic events in this population was determined. The charts of all additional gravid women with antepartum or postpartum deep vein thrombosis or pulmonary embolism diagnosed during the study period were also reviewed. The prevalence of these disorders among the pregnant population for whom extended bed rest was not prescribed as part of the treatment of premature labor or preterm premature rupture of membranes was also calculated. Statistical comparison of the prevalences in the 2 populations was undertaken by means of the chi(2) analysis with the Fisher exact test. RESULTS: There were 192 patients admitted during the study period who had extended bed rest prescribed as part of the treatment of premature labor or preterm premature rupture of membranes. Three of these women had thromboembolic events, for a prevalence of 15.6 cases per 1000 women. Five additional gravid women were admitted for the treatment of deep vein thrombosis or pulmonary embolism. There were 6164 deliveries among women not treated with extended bed rest for premature labor or preterm premature rupture of membranes during this period. Thus the prevalence of these phenomena among the remaining pregnant women was 0.8 cases per 1000 women. The prevalences of these disorders in the 2 populations were highly significantly different. CONCLUSION: The prevalence of thromboembolic events among women for whom extended bed rest is prescribed as part of the treatment of premature labor or preterm premature rupture of membranes is significantly increased with respect to that among gravid women who do not receive this therapy and is substantially higher than previously reported. If this finding is confirmed in other populations, it may be prudent to undertake further studies to determine whether this prevalence can be reduced. PMID- 10819837 TI - In vitro effects of sildenafil and phentolamine, drugs used for erectile dysfunction, on human sperm motility. AB - OBJECTIVE: The aim of this study was to determine the effects of sildenafil and phentolamine on sperm motility in vitro. STUDY DESIGN: Semen or washed sperm was mixed with various doses of sildenafil or phentolamine and analyzed for motility during a 30-minute period. The pH was measured for each of the samples tested. Statistical analyses were performed with analysis of variance. RESULTS: A 200 microg/mL dose of sildenafil had no effect on sperm motility. However, the highest dose (2000 microg/mL) significantly reduced motility by about 50%. The pH was reduced in this high-dose sample. The lowest dose of phentolamine (20 microg/mL) had no effect, whereas a dose of 200 microg/mL resulted in a significant reduction in sperm motility. The highest dose (2000 microg/mL) stopped virtually all sperm from moving. CONCLUSION: This study demonstrated a direct dose-related effect of phentolamine on reducing sperm motility. Only the highest dose of sildenafil had an effect, which may have been caused by a decline in pH. PMID- 10819838 TI - The effect of diabetes on alpha2-adrenergic receptor activity in the reproductive centers of the female rat brain. AB - OBJECTIVE: We hypothesized that known diabetes-induced deficits in female rat reproduction may result in part from decreased central alpha(2)-noradrenergic receptor density or affinity. STUDY DESIGN: Female rats were oophorectomized and divided into 2 groups; one group received streptozocin during the operation to induce diabetes, and the other served as a nondiabetic control group. Random blood glucose levels were measured. Half the rats in each group were killed on postoperative day 10, and half were killed on postoperative day 14. Direct radioligand binding assays were performed on tissue prepared from the hypothalamus, preoptic area, and cortex of each rat. Analysis of variance was used to evaluate intergroup differences in receptor concentration or equilibrium constant. RESULTS: We detected no significant difference in the mean receptor concentration or equilibrium constant between the groups with and without diabetes in the hypothalamus, the preoptic area, and the cortex on postoperative day 10 or 14. CONCLUSION: Diabetes-induced impairments in female rat reproduction do not involve alterations in alpha(2)-receptor density or affinity in the hypothalamus, preoptic area, or cortex. PMID- 10819839 TI - The effect of raloxifene on the uterine weight response in immature mice exposed to 17beta-estradiol, 1,1,1-trichloro-2, 2-bis(p-chlorophenyl)ethane, and methoxychlor. AB - OBJECTIVE: The purpose of this study was to determine the effects of raloxifene on the uterine responses to both estradiol and the environmental estrogens 1,1,1 trichloro-2, 2-bis(p-chlorophenyl)ethane and methoxychlor in immature mice. STUDY DESIGN: Immature female mice received the following compounds alone or in combination: sesame oil (control), 17beta-estradiol 1 mg/kg body weight, tamoxifen 1 mg/kg body weight, raloxifene 5 mg/kg body weight, 1,1,1-trichloro 2,2-bis(p-chlorophenyl)ethane 10 mg/kg body weight, and methoxychlor 10 mg/kg body weight. The animals were treated subcutaneously once a day for 5 consecutive days with the compound or compounds of interest in 0.1 mL sesame oil. Approximately 24 hours after the final treatment the animals were killed and the uteri were excised, stripped of remaining fat and mesentery, and weighed. Groups were analyzed with analysis of variance. RESULTS: Estradiol increased the mean (+/-SE) weight from 20 +/- 6.4 mg (as measured in the control group) to 77 +/- 6.2 mg. Tamoxifen increased uterine weight to 60 +/- 6.2 mg; however, raloxifene had no effect on uterine weight. Both 1,1,1-trichloro-2, 2-bis(p chlorophenyl)ethane and methoxychlor increased uterine weight significantly, to 82 +/- 2.4 mg and 35 +/- 6.0 mg respectively. When raloxifene was coadministered with 17beta-estradiol it did not block the increase in uterine weight; however, when raloxifene was coadministered with 1,1,1-trichloro-2, 2-bis(p chlorophenyl)ethane or methoxychlor, it completely blocked the uterine weight gain induced by either xenoestrogen. CONCLUSION: Raloxifene blocked the xenoestrogens 1,1,1-trichloro-2, 2-bis(p-chlorophenyl)ethane and methoxychlor but did not block 17beta-estradiol in the mouse model described. These results suggest that the xenoestrogens exert their estrogenic activities through a different site on the estrogen receptor or through a different mechanism than 17beta-estradiol. PMID- 10819840 TI - Fetal fibronectin detection as a predictor of preterm birth in actual clinical practice. AB - OBJECTIVE: This study was undertaken to determine whether fetal fibronectin determination is more useful for predicting preterm delivery in clinical practice than it has appeared to be in prospective blinded studies. STUDY DESIGN: Charts of 151 patients with fetal fibronectin tests performed during 2 years were reviewed. Patients were included if they had symptoms of preterm labor, a singleton pregnancy at 24 to 35 weeks' gestation, intact membranes, and cervical dilatation < or =3 cm. RESULTS: Complete data were available for 85 tests. For delivery within 7 days after specimen collection the sensitivity, specificity, positive predictive value, and negative predictive value were 89%, 84%, 40%, and 98%, respectively. The positive predictive value was greater (P <.002) than those reported in three prospective studies evaluating delivery within 7 days in patients with symptoms. Gestational age at delivery and birth weight were lower for patients with positive results (P <. 0001 and P <.006, respectively). Patients with positive results were also treated more with tocolysis, corticosteroid use, and hospitalization than were patients with negative results. For direct comparison with studies of patients with cervical dilatation <3 cm, only 4 patients with cervical dilatation of 3 cm were enrolled. All 4 had negative results of fetal fibronectin testing, and their outcomes therefore did not affect the positive predictive value. CONCLUSION: The positive predictive value of fetal fibronectin measured in actual clinical practice was significantly greater for delivery within 7 days than has been reported in blinded prospective studies. PMID- 10819841 TI - Nucleated red blood cell count in the differentiation of fetuses with pathologic growth restriction from healthy small-for-gestational-age fetuses. AB - OBJECTIVE: The purpose of this study was to evaluate the utility of the neonatal nucleated red blood cell count in differentiating the fetus with growth restriction from the small but otherwise healthy fetus. STUDY DESIGN: Perinatal outcomes were evaluated prospectively for all neonates admitted to the neonatal intensive care unit in 1997. Nonanomalous neonates with normal phenotype and a complete blood cell count performed within the first 6 hours after birth were included in the study. All neonates with birth weights lower than the 10th percentile for gestational age were considered small for gestational age. Neonates were divided into four groups: small-for-gestational-age neonates with elevated nucleated red blood cell counts, appropriately grown neonates with elevated nucleated red blood cell counts, small-for-gestational-age neonates with normal nucleated red blood cell counts, and appropriately grown neonates with normal nucleated red blood cell counts. Analysis of variance, chi(2) tests, and stepwise regression were used for statistical analysis. RESULTS: Two hundred thirty-seven neonates met the inclusion criteria. Forty-three were small for gestational age. Small-for-gestational-age neonates with high nucleated red blood cell counts had significantly lower umbilical artery pH and were more likely to require mechanical ventilation or blood pressure support agents. Subgroup analysis demonstrated that small-for-gestational-age neonates with elevated nucleated red blood cell counts had significantly more adverse outcomes than did small-for-gestational-age neonates with normal nucleated red blood cell counts. Outcomes of small-for-gestational-age neonates with normal nucleated red blood cell counts were essentially identical to those of appropriately grown neonates. CONCLUSION: An elevated nucleated red blood cell count may distinguish the fetus with growth restriction from the small but healthy fetus. PMID- 10819842 TI - Differences in TDx fetal lung maturity assay values between twin and singleton gestations. AB - OBJECTIVE: The purpose of this study was to quantify differences in indexes of pulmonary maturity between singleton and twin gestations by means of the TDx fetal lung maturity assay. STUDY DESIGN: We identified records of a total of 830 singleton and twin pregnancies not complicated by diabetes and delivered between 28 and 37 weeks' gestation from December 1994 through August 1995. Among these, 170 (20%) had TDx fetal lung maturity measurements performed within 72 hours of delivery. Linear regression was used to assess differences in TDx fetal lung maturity assay values between singleton gestations (n = 143 gestations) and twin gestations (n = 27 gestations) while controlling for potential confounding factors. RESULTS: Twin gestations were no more likely than singleton gestations to undergo TDx fetal lung maturity screening (odds ratio, 1.3; 95% confidence interval, 0.8-2.2). Pregnancy complications and corticosteroid treatment were similar in the two groups. After 31 weeks' gestation the twin gestations had significantly higher TDx fetal lung maturity values. Linear regression with controls for gestational age indicated that twin gestations on average had a TDx fetal lung maturity value that was 22.0 mg/g (95% confidence interval, 9.8-34.6 mg/g) higher than that of gestational age-matched singleton gestations. CONCLUSION: Beyond 31 weeks' gestation twin pregnancies appeared to have a TDx fetal lung maturity value that was 22 mg/g higher than that of singleton pregnancies. If the underlying incidences of respiratory distress syndrome are similar between twin and singleton gestations, then the potential exists for false-positive prediction of adequate lung maturity values among twin gestations. PMID- 10819843 TI - Outcomes of extremely low-birth-weight infants between 500 and 750 g. AB - OBJECTIVE: The purpose of this study was to describe morbidity, mortality, and developmental outcomes among extremely low-birth-weight infants (500-750 g). STUDY DESIGN: This retrospective cohort study included 167 live-born infants born at Long Beach Memorial Medical Center between January 1990 and December 1995. RESULTS: Mortality rates were 86% among infants <600 g and 44% among those > or =600 g, and the rate decreased with increasing gestational age. The absence of chorioamnionitis (P =.01) and the use of antepartum corticosteroids (P <.0001) or neonatal surfactant (P =.0001) were associated with survival. Sixty-four percent of studied infants had respiratory distress syndrome, and 17% had grade III or IV intraventricular hemorrhage. Among the 63 survivors 57% were tested at 30 months of corrected age with the Bayley Scales of Infant Development. Mild or significant delays were seen on the mental development index in 68% of these cases and on the psychomotor development index in 58% of cases. CONCLUSION: Morbidity and mortality rates and the rate of developmental delay among infants with birth weights between 500 and 750 g were significant. PMID- 10819844 TI - Estimation of fetal weight before and after amniotomy in the laboring gravid woman. AB - OBJECTIVE: This study was undertaken to search for differences between fetal weights estimated both ultrasonographically and clinically before and after amniotomy in laboring gravid women. STUDY DESIGN: Estimates of fetal weight (ultrasonographic and clinical) were obtained for laboring gravid women before and after amniotomy. These estimates were compared with actual birth weights determined post partum. RESULTS: One hundred sixty-two patients completed the study protocol. Comparisons made with unpaired Student t test analyses demonstrated a difference (P <.001) between ultrasonographically estimated fetal weights before and after amniotomy. Simple regression analysis showed a correlation between both ultrasonographic and clinical estimates of fetal weight and actual birth weights before and after amniotomy, with postamniotomy clinical estimates having the strongest correlation (ultrasonographic preamniotomy estimate, R = 0.717; ultrasonographic postamniotomy estimate, R = 0.630; clinical preamniotomy estimate, R = 0.742; and clinical postamniotomy estimate, R = 0.788). Of all ultrasonographic parameters measured, preamniotomy abdominal circumference correlated best with actual birth weight (R = 0.730). CONCLUSION: Clinical estimates of fetal weight after amniotomy correlated well with actual birth weights. Preamniotomy abdominal circumference was the ultrasonographic parameter best for prediction of actual birth weight. Maternal weight affected clinical but not ultrasonographic estimates of fetal weight in this study. However, clinical estimates of fetal weight were actually superior to ultrasonographic estimates of fetal weight in this study. PMID- 10819845 TI - Drugs for the gynecologist to prescribe in the prevention of breast cancer: current status and future trends. AB - Tamoxifen was approved for breast cancer prevention in October 1998. Thus, for the first time, we as gynecologists are being asked to prescribe this drug to healthy women. In the past each one of us has cared for women with breast cancer who have been treated with tamoxifen by oncologists or breast surgeons for the malignancy. Effects of tamoxifen on the uterus resulting in carcinomas, hyperplasia, and polyps are well known. Furthermore, tamoxifen has estrogenic properties in the venous system, increasing the incidence of deep vein thrombosis and pulmonary emboli. A new SERM (selective estrogen receptor modulator), raloxifene, has been approved for prevention and treatment of osteoporosis in postmenopausal women. It does not have stimulatory effects on the endometrium; however, it is estrogenic in the venous system. Preclinical data, as well as the breast cancer incidence reported in studies of the skeleton, seem to indicate that its effects in the breast are similar to those of tamoxifen. This article reviews tamoxifen and the new SERM, raloxifene, in an attempt to help gynecologists better understand each compound and what data are currently known, what we hope to learn from future studies, and what currently makes sense for clinical practice. PMID- 10819846 TI - Changes in hemodynamic parameters and volume homeostasis with the menstrual cycle among women with a history of preeclampsia. AB - OBJECTIVE: Among women with a history of preeclampsia the prevalence of hemodynamic and clotting disorders is elevated. In this study we tested the hypothesis that the normal cyclic variation in hemodynamic and renal function parameters with the menstrual cycle that is seen among healthy women would be preserved in women with a history of preeclampsia irrespective of whether they had an underlying hemodynamic or clotting disorder. STUDY DESIGN: We compared the hemodynamic and volume cyclic variations during the menstrual cycle among women with a history of preeclampsia (n = 39) with those among healthy parous control women (control group, n = 10). The participants with a history of preeclampsia were subdivided into groups of women with hypertension with or without thrombophilia (hypertension group, n = 10), women with a normotension and a thrombophilic disorder (thrombophilia group, n = 17), and women without either of these abnormalities (symptom-free group, n = 12). We measured > or =5 months post partum, once during the follicular phase of the menstrual cycle (day 5 +/- 2) and once during the luteal phase (day 22 +/- 2), the following variables: body weight and length, mean arterial pressure, heart rate, cardiac output, plasma volume, glomerular filtration rate, effective renal plasma flow, and concentrations of renal volume homeostatic hormones, reproductive hormones, and catecholamines. From the measured data we calculated body mass index, cardiac index, left ventricular work, total peripheral and renal vascular resistances, effective renal blood flow, and renal filtration fraction. RESULTS: The hypertension group differed from the control group in having higher baseline (follicular phase) values for cardiac output, cardiac output, left ventricular work, renal vascular resistance, and atrial natriuretic peptide and norepinephrine levels. The symptom free group differed from the control group in having a lower baseline plasma volume and higher baseline cardiac output and left ventricular work values. Women in the thrombophilia group were comparable to those in the control group with respect to baseline hemodynamic and renal function variables except for a higher renal vascular work value. In the control group heart rate, plasma volume, effective renal plasma volume, effective renal blood flow, and concentrations of renin-angiotensin-aldosterone system hormones and norepinephrine were increased during the luteal phase with respect to values during the follicular phase, whereas the renal vascular resistance and atrial natriuretic peptide values were decreased. In the three subgroups of women with a history of preeclampsia this cyclic pattern with the menstrual cycle was preserved for most of these parameters. CONCLUSION: Although baseline hemodynamic and volume status among women with a history of preeclampsia differed from that among healthy parous control subjects, the cyclic variation with the menstrual cycle was largely preserved. PMID- 10819847 TI - Laser scanning confocal microscopy of cervical tissue before and after application of acetic acid. AB - OBJECTIVES: The use of high-resolution in vivo confocal imaging may offer a clinically useful adjunct to standard methods for the diagnosis and screening of epithelial precancers. This study assesses the feasibility of real-time confocal reflectance imaging of cervical tissue and the use of acetic acid as a contrast agent to increase visualization of cell nuclei. STUDY DESIGN: A confocal microscope was used to image cervical cells and colposcopically normal and abnormal cervical biopsy specimens. Images were obtained before and after the application of 6% acetic acid. RESULTS: The confocal imaging system resolved subcellular detail throughout the entire epithelial thickness. Normal and abnormal tissues were clearly able to be differentiated. Addition of acetic acid enhanced nuclear signal in all acquired images. CONCLUSION: High-contrast reflected light images of cervical tissue are attainable in near real time. Acetic acid significantly increases light scattering from cell nuclei, which may partially explain why acetowhitening occurs. PMID- 10819848 TI - Effects of leuprolide acetate on low-grade endometrial stromal sarcoma. AB - We describe a low-grade endometrial stromal sarcoma coexistent with leiomyoma and adenomyosis treated with leuprolide acetate. We describe its histologic characteristics and clinical significance. PMID- 10819849 TI - Gonadotropin-releasing hormone analog repairs reduced endometrial cell apoptosis in endometriosis in vitro. AB - OBJECTIVE: Impaired sensitivity of endometrial tissue to spontaneous apoptosis in women with endometriosis contributes to the abnormal implantation and growth of endometrium at ectopic sites. Our purpose was to examine the effect of gonadotropin-releasing hormone analog, widely used in the treatment of endometriosis, on the reduced rate of endometrial apoptosis in endometriosis. STUDY DESIGN: Paired ectopic and eutopic endometrial tissue specimens were obtained from 13 patients with endometriosis, and control samples were taken from 8 patients with uterine myoma. Apoptotic cell death was assessed biochemically and morphologically with an enzyme-linked immunoassay and Hoechst No. 33342 staining of deoxyribonucleic acid fragment, respectively. RESULTS: Spontaneous apoptosis was significantly lower in ectopic and eutopic endometrial tissue from patients with endometriosis (0.22 +/- 0.082 in absorbance) than in endometrial tissue from control subjects (0.52 +/- 0.483)(P < 0.001). Incubation with a gonadotropin-releasing hormone analog (1 micromol/L) increased the apoptotic rate of endometrial cells from patients with endometriosis to 0.56 +/- 0.501 (P <.001). The effect of this gonadotropin-releasing hormone revealed a dose dependency; a half-maximal effect occurred with 10 nmol/L; however, the control endometrium was not affected. CONCLUSION: Exposure to gonadotropin-releasing hormone results in changes of the sensitivity of endometriotic endometrium to spontaneous apoptosis; these changes in sensitivity may, in turn, release endometrial cells from resistance to apoptosis and result in reduced survival and growth. This phenomenon could, at least in part, account for the therapeutic action of gonadotropin-releasing hormone analog on endometriosis. PMID- 10819850 TI - Puerperal infection after cesarean delivery: evaluation of a standardized protocol. AB - OBJECTIVE: Our goal was to evaluate an antibiotic protocol for treatment of postcesarean endometritis. STUDY DESIGN: Endometritis was diagnosed as a persistent fever > or =100.4 degrees F beyond 24 hours after cesarean delivery and one or more of the following: uterine tenderness, tachycardia, foul lochia, or leukocytosis. Antibiotic therapy included gentamicin plus clindamycin and ampicillin (or vancomycin) as a triple antimicrobial in 148 women. Antibiotic failure was defined as persistent fever after 5 days of antibiotics and 72 hours of triple antibiotics. RESULTS: Between 1993 and 1996, 322 of 1643 (20%) women were diagnosed with postcesarean endometritis. One hundred seventy-four patients (54%) were cured with clindamycin-gentamicin, and 129 who additionally received ampicillin or vancomycin (40%) were cured. Nineteen of the 322 (6%) women had persistent fever despite triple antibiotics. Of these, 6 had a wound complication, 12 were suspected to have antimicrobial resistance, and 1 had an infected hematoma. CONCLUSION: A prospective protocol consisting of clindamycin gentamicin plus the selective addition of ampicillin or vancomycin cured 303 of 322 (94%) women with postcesarean endometritis. PMID- 10819851 TI - Unstable angina during pregnancy in two patients with premature coronary atherosclerosis and aortic stenosis in association with familial hypercholesterolemia. AB - Obstructive atherosclerotic coronary artery disease is uncommon in women during childbearing age, and the occurrence of myocardial ischemia during pregnancy has therefore been anecdotal. Two young patients with premature coronary artery disease in association with familial hypercholestrolemia had unstable angina in the second trimester of pregnancy. Workup revealed coronary artery disease and aortic stenosis. One patient opted for abortion at the twentieth week of gestation, and the other decided to continue pregnancy and was delivered by cesarean at 28 weeks' gestation. Coronary artery bypass grafting was performed after pregnancy in both patients. In addition, one of the patients underwent aortic valve replacement, and other had replacement of the narrowed ascending aorta with uneventful recovery. Our report describes an uncommon presentation of unstable angina during pregnancy in 2 young women with premature coronary artery disease and aortic valvular and supravalvular stenosis as a result of familial hypercholesterolemia. The management of these conditions during pregnancy is influenced by the effects of available therapeutic modalities on both maternal and fetal outcome. PMID- 10819852 TI - Identification of monozygotic twins that are concordant for preeclampsia. AB - Preeclampsia is thought to be discordant in monozygotic twins. While recruiting for a genetic study into preeclampsia, we identified 4 sets of twins and a triplet gestation; all were monozygous on deoxyribonucleic acid "fingerprinting." Two twins were concordant for preeclampsia, and 2 of the triplets had pregnancy induced hypertension, although only 1 of them had proteinuria. Hence we confirm the existence of monozygotic twins that are concordant for preeclampsia. PMID- 10819853 TI - Cost-effectiveness of a delayed pushing policy for patients with epidural anesthesia. The PEOPLE (Pushing Early or Pushing Late with Epidural) Study Group. AB - OBJECTIVE: The purpose of this study was to estimate the economic efficiency of a policy of delayed pushing for nulliparous women who have full dilatation while they are under epidural anesthesia. STUDY DESIGN: A cost-effectiveness evaluation was based on a randomized controlled trial. Resource use and clinical effectiveness data were collected for 1862 women who were randomly allocated to either a delayed pushing group (n = 936) or an early pushing group (n = 926). Costs (in 1997 Canadian dollars) were collected for each item of resource use. Sensitivity analysis was used to examine the robustness of the main results. RESULTS: Delayed pushing was effective at reducing the number of difficult deliveries (relative risk, 0.79; 95% confidence interval, 0.66 to 0.95). The mean cost of intrapartum care was significantly higher in the delayed pushing group ($625.86 vs $557.64; P <.0005). There were no significant differences in mean costs of postnatal care ($2146.67 vs $2133.54; P =.871) or total hospital care ($2772.53 vs $2691.18; P =.324). The incremental cost per difficult delivery prevented was estimated at $1743.06. The incremental cost-effectiveness ratio remained fairly robust with variations in the values of key parameters incorporated into the sensitivity analysis. CONCLUSIONS: The results of this economic evaluation should inform decision makers determining whether to advocate a policy of delayed pushing for nulliparous women who have full dilatation while they are under epidural anesthesia. PMID- 10819854 TI - Multicenter, randomized, controlled trial of delayed pushing for nulliparous women in the second stage of labor with continuous epidural analgesia. The PEOPLE (Pushing Early or Pushing Late with Epidural) Study Group. AB - OBJECTIVE: This study was undertaken to determine whether a policy of delayed pushing for nulliparous women with continuous-infusion epidural analgesia reduces the risk of difficult delivery (cesarean delivery, operative delivery from a midpelvic position, low-pelvic procedures with rotation >45 degrees ). STUDY DESIGN: In this multicenter, randomized, controlled trial women in the delayed pushing group (n = 936) were advised to wait > or =2 hours after full dilatation before commencement of pushing. Women in the early pushing group (n = 926) were advised to commence pushing as soon as they had been randomly assigned. RESULTS: Difficult delivery was reduced with delayed pushing (relative risk, 0.79; 95% confidence interval, 0.66-0.95). The greatest effect was on midpelvic procedures (relative risk, 0.72; 95% confidence interval, 0.55-0.93). Although there was little evidence for an effect on low-pelvic procedures, spontaneous delivery was more frequent among women who practiced delayed pushing (relative risk, 1.09; 95% confidence interval, 1.00-1.18). Abnormal umbilical cord blood pH (<7.15 venous value or <7.10 arterial value) was more frequent in the delayed pushing group (relative risk, 2.45, 95% confidence interval, 1.35-4. 43). However, scores for a summary indicator, the Neonatal Morbidity Index, were similarly distributed in the 2 groups. CONCLUSION: Delayed pushing is an effective strategy to reduce difficult deliveries among nulliparous women. PMID- 10819855 TI - An oxytocin receptor antagonist (atosiban) in the treatment of preterm labor: a randomized, double-blind, placebo-controlled trial with tocolytic rescue. AB - OBJECTIVES: This study was designed to evaluate the efficacy and safety of the oxytocin receptor antagonist atosiban in the treatment of preterm labor. STUDY DESIGN: A multicenter, double-blind, placebo-controlled trial with tocolytic rescue was designed. Five hundred thirty-one patients were randomized to receive, and 501 received, either intravenous atosiban (n = 246) or placebo (n = 255), followed by subcutaneous maintenance with the assigned agent. Standard tocolytics as rescue tocolysis were permitted after 1 hour of either placebo or atosiban if preterm labor continued. The primary end point was the time from the start of study drug to delivery or therapeutic failure. Secondary end points were the proportion of patients who remained undelivered and did not receive an alternate tocolytic at 24 hours, 48 hours, and 7 days. RESULTS: No significant difference was found in the time from start of treatment to delivery or therapeutic failure between atosiban and placebo (median, 25.6 days vs 21.0 days, respectively; P =.6). The percentages of patients remaining undelivered and not requiring an alternate tocolytic at 24 hours, 48 hours, and 7 days were significantly higher in the atosiban group than in the control group (all P < or =.008). A significant treatment-by-gestational age interaction existed for the 48-hour and 7-day end points. Atosiban was consistently superior to placebo at a gestational age of > or =28 weeks. Fourteen atosiban-treated patients and 5 placebo-treated patients were randomized at <24 weeks; the incidence of fetal-infant deaths was higher for the atosiban group at <24 weeks. Maternal-fetal adverse events were similar except for injection-site reactions, which occurred more often with atosiban. CONCLUSIONS: In this trial the treatment of patients in preterm labor with atosiban resulted in prolongation of pregnancy for up to 7 days for those at a gestational age > or =28 weeks, and this occurred with a low rate of maternal fetal adverse effects. In addition, at a gestational age > or =28 weeks, the infant morbidity and mortality of atosiban-initiated standard care were similar to those with placebo-initiated standard care. Given that all patients in this study were eligible for tocolysis and that, in practice, nearly all patients who are eligible for a tocolytic receive one, the benefit of using atosiban is the placebo-like maternal-fetal side effect profile. These observations support the use of this oxytocin receptor antagonist in the treatment of patients in preterm labor with intact membranes. Efficacy and infant outcome data at <28 weeks are inconclusive. PMID- 10819856 TI - Maintenance treatment of preterm labor with the oxytocin antagonist atosiban. The Atosiban PTL-098 Study Group. AB - OBJECTIVES: Patients admitted with an acute episode of preterm labor who respond to early intravenously administered tocolysis remain at risk of having subsequent episodes of preterm labor and preterm delivery. Several pharmacologic agents have been used in an attempt to reduce subsequent episodes of preterm labor, and all are associated with significant side effects. Atosiban, an oxytocin receptor antagonist, is effective in the treatment of an acute episode of preterm labor. This study was designed to compare the efficacy and safety of atosiban with those of placebo maintenance therapy in women with preterm labor who achieved uterine quiescence with intravenous atosiban. STUDY DESIGN: A multicenter, double-blind, placebo-controlled trial was designed for patients in preterm labor who responded to early intravenous treatment with atosiban. Five hundred thirteen patients were randomly assigned to receive maintenance therapy, 252 to receive atosiban, and 251 to receive matching placebo. Maintenance therapy was administered as a continuous subcutaneous infusion, via pump, of 30 microg/min to the end of 36 weeks' gestation. The primary end point was the number of days from the start of maintenance therapy until the first recurrence of labor. A secondary end point was the percentage of patients receiving subsequent intravenous atosiban therapy. RESULTS: The time (median) from the start of maintenance treatment to the first recurrence of labor was 32.6 days with atosiban and 27.6 days with placebo (P =.02). At least one subsequent intravenous atosiban treatment was needed by 61 atosiban patients (23%) and 77 placebo patients (31%). Except for injection site reactions, adverse event profiles of atosiban and placebo were comparable. There were 4 neonatal deaths reported in the atosiban group and 5 in the placebo group after the start of maintenance therapy. Infant outcomes (including birth weight) were comparable between maintenance and treatment groups. CONCLUSIONS: Maintenance therapy with the oxytocin receptor antagonist atosiban can prolong uterine quiescence after successful treatment of an acute episode of preterm labor with atosiban. Treatment was well tolerated. PMID- 10819857 TI - Double-blind, randomized, controlled trial of atosiban and ritodrine in the treatment of preterm labor: a multicenter effectiveness and safety study. AB - OBJECTIVE: This study was undertaken to compare the efficacy and safety of intravenous administration of atosiban versus ritodrine for the treatment of preterm labor. STUDY DESIGN: Women with preterm labor and intact membranes diagnosed at 23 to 33 gestational weeks (n = 247) were randomly assigned to treatment arms and received atosiban (6.75 mg intravenous bolus, 300 microg/min for 3 hours, then 100 microg/min intravenously) or ritodrine (0.10-0.35 mg/min intravenously) for as long as 18 hours. Tocolytic effectiveness was assessed in terms of the numbers of women who had not been delivered after 48 hours and after 7 days. Safety was assessed in terms of maternal side effects and neonatal morbidity. Secondary outcomes included mean gestational age at delivery and mean birth weight. An intent-to-treat analysis was performed with the Cochran-Mantel Haenszel test. RESULTS: The proportion of women who had not been delivered at 48 hours was 84.9% (n = 107) in the atosiban group and 86.8% (n = 105) in the ritodrine group. At 7 days 92 women had still not been delivered in both the atosiban (73.0%) and ritodrine (76.0%) groups. Neither of these differences was statistically significant. The incidence of maternal cardiovascular side effects was substantially lower in the atosiban group (4.0% vs 84.3%, P <.001). In addition, intravenous therapy was terminated more frequently as a result of maternal adverse events in the ritodrine group (29.8%) than in the atosiban group (0.8%). The overall occurrences of fetal adverse events in the two treatment groups were comparable. Neonatal morbidity was similar between the treatment groups after adjustment for unbalanced enrollment of women with multiple pregnancies and for gestational ages within treatment groups. CONCLUSION: Atosiban was comparable in clinical effectiveness to conventional ritodrine therapy but was better tolerated than ritodrine, with no evidence of significant maternal or fetal adverse events. Neonatal morbidity, which was similar between the two treatment arms, was apparently related to the gestational age of the infant rather than to the exposure to either tocolytic agent. PMID- 10819858 TI - Plasma adenosine levels increase in women with normal pregnancies. AB - OBJECTIVE: The aim of this study was to investigate plasma adenosine levels during normal pregnancy and to evaluate the possible roles of platelet activation and 5'-nucleotidase as causes of changes in adenosine levels. STUDY DESIGN: We measured plasma adenosine levels, the platelet activation markers beta thromboglobulin and platelet factor 4, and 5'-nucleotidase activity, which catalyzes dephosphorylation from adenosine monophosphate to adenosine, in 34 nonpregnant women and 34 women with normal pregnancies in the third trimester. RESULTS: The mean plasma adenosine level in pregnant women was 0.59 +/- 0.08 micromol/L (mean +/- SEM), which was significantly higher than that found in nonpregnant women (0.18 +/- 0.04 micromol/L; P <.01). In pregnant women plasma beta-thromboglobulin levels, platelet factor 4 levels, and 5'-nucleotidase activity were significantly higher than in nonpregnant women (P <.05). CONCLUSION: The increase of plasma adenosine may be attributed at least in part to platelet activation and an increase of 5'-nucleotidase activity during normal pregnancy. This increase may be an endogenous compensatory mechanism that diminishes platelet activation and maintains vessel integrity during normal pregnancy. PMID- 10819859 TI - Clinical significance of early diastolic notch depth: uterine artery Doppler velocimetry in the third trimester. AB - OBJECTIVE: Our purpose was to evaluate the usefulness of early diastolic notch depth in predicting adverse perinatal outcome. STUDY DESIGN: Uterine artery Doppler velocimetry is widely applied in predicting pregnancy outcomes. Recent studies of uterine artery early diastolic notch have demonstrated its usefulness as a marker for fetal well-being. The early diastolic notch represents the reflected blood flow of uteroplacental circulation. This study was carried out under the hypothesis that when early diastolic notch is present evaluating its depth plays a significant role in predicting poor pregnancy outcomes. Its ability to predict adverse perinatal outcomes was evaluated in 198 pregnant women with early diastolic notch after 28 weeks' gestation. RESULTS: The incidence of adverse perinatal outcome (5-minute Apgar score <7, cesarean delivery because of fetal distress, admission to neonatal intensive care unit, fetal growth restriction, or perinatal death) was 90.5% when the notch index was /=0.90, the respective incidences were 72.3%, 46. 9%, and 28.1%. CONCLUSION: This study suggests that evaluating notch depth in the presence of early diastolic notch on uterine artery velocimetry during the third trimester would be a useful method for predicting adverse perinatal outcomes. PMID- 10819860 TI - Outcome after successful resuscitation of babies born with apgar scores of 0 at both 1 and 5 minutes. AB - OBJECTIVE: Our purpose was to evaluate the outcome of infants who underwent successful resuscitation after initial Apgar scores of 0 at both 1 and 5 minutes. STUDY DESIGN: Eligible infants were identified through the perinatal database at the University of Tennessee, Memphis. Hospital records and long-term outcomes, where available, of babies who met the above criteria occurring between January 1986 and February 1999 were reviewed. RESULTS: Thirty-three of 81,603 infants (0.4/1000 births) met our study criteria. Twenty-two (67%) babies died during hospitalization. Mortality decreased significantly from 100% for babies with a birth weight of <750 g to 38% for those weighing > or =2500 g at birth (P =.03). All 6 babies delivered before 26 weeks' gestation died. The incidence of 10 minute Apgar scores >0 was significantly higher among survivors than among those who subsequently died (82% vs 33%, P <.05). Nine survivors had hypoxic-ischemic encephalopathy diagnosed before discharge. Of the 7 infants with available follow up, 4 had significant persisting morbidity. Two infants had normal neurologic examinations at follow-up. CONCLUSION: Survival in babies born with 1- and 5 minute Apgar scores of 0 is predicted by birth weight, gestational age, and 10 minute Apgar score. Long-term sequelae are common but not ensured. PMID- 10819861 TI - Obstetric emergencies precipitated by malignant brain tumors. AB - OBJECTIVE: Our goal was to present a case series of pregnancy-associated malignant brain tumors. STUDY DESIGN: A review was conducted from 1978-1998 at 5 hospitals. RESULTS: Ten women were diagnosed with a malignant brain tumor during pregnancy (n = 8) or post partum (n = 2). Patients diagnosed antenatally exhibited severe symptoms, manifest between 27 and 32 weeks' gestation. Six were emergently delivered of their infants because of maternal deterioration, and 2 were delivered electively in the early third trimester after documentation of fetal pulmonary maturity. There were 4 maternal deaths and 1 neonatal death; all of the other infants maintained viability. CONCLUSIONS: Malignant brain tumors rarely occur in pregnancy. In contrast to reports that describe an indolent course, each of the 8 antenatal patients experienced a neurologic crisis. If symptoms are amenable to pharmacologic control, we advocate delivery in the early third trimester after documentation of fetal pulmonary maturity. To minimize temporal lobe or cerebellar herniation in neurologically unstable patients, a consideration should be made for cesarean delivery with the patient under general anesthesia, followed by immediate neurosurgical decompression. PMID- 10819862 TI - Splenic artery Doppler peak systolic velocity predicts severe fetal anemia in rhesus disease. AB - OBJECTIVE: We sought to determine whether main splenic artery Doppler peak systolic velocity predicts severe anemia in the rhesus-alloimmunized fetus. STUDY DESIGN: Splenic artery Doppler peak systolic velocity was obtained before cordocentesis in rhesus-alloimmunized fetuses. Normative values for mean peak systolic velocity based on gestational age were obtained cross-sectionally from a separate group of 144 normal fetuses. The peak systolic velocity values in the study group were expressed as multiples of the median for gestation, and threshold values were used as a screening test for severe anemia. The hemoglobin deficit was defined as mean hemoglobin for gestation minus measured hemoglobin. A hemoglobin deficit value of > or =5 g/dL was used to define severe anemia. We used the peak systolic velocity to screen for severe anemia in the overall study group and the subgroups with or without prior transfusions. RESULTS: The study population consisted of 26 singleton nonhydropic fetuses in which cordocentesis and Doppler measurements were performed on a total of 55 occasions. The mean gestational age and standard deviation at cordocentesis was 29.6 +/- 4.0 weeks. Severe anemia was noted in 20% of fetal cord blood specimens obtained. On the basis of a receiver operating characteristic curve, a peak systolic velocity of > or =1.4 multiples of the median had a detection rate of 100%, with a false positive rate of 20.8% in the subgroup with no prior transfusion (relative risk, 4.8; 95% confidence interval, 2.2-10.5). For peak systolic velocity threshold of > or =1.50 multiples of the median, corresponding values in the group with one prior transfusion were 80% and 12.5%, respectively (relative risk, 2.5; 95% confidence interval, 1.2-5.3). There was no risk of severe anemia with a peak systolic velocity below the median for gestation. CONCLUSION: Fetal hydrops is rare, with a hemoglobin deficit of <5 g/dL. In the first such report the main splenic artery peak systolic velocity was noted to be a strong predictor of severe anemia. For the overall population, all such instances could be diagnosed while cordocentesis was performed 22.7% of the time. There is no risk of severe anemia with Doppler peak systolic velocities below the median for gestational age. The measurement is easily obtained and should be investigated as a clinical tool for minimizing the necessity for cordocentesis. PMID- 10819863 TI - Power spectral analysis of R-R interval variability before and during the sinusoidal heart rate pattern in fetal lambs. AB - OBJECTIVE: The appearance of the sinusoidal heart rate pattern found on fetal cardiotocograms has not been fully explained, either physiologically or clinically. In this study we performed power spectral analysis on the sinusoidal heart rate pattern obtained by administration of arginine vasopressin and atropine sulfate to investigate its frequency components in fetal lambs with long term instrument implantation. STUDY DESIGN: Eleven tests were performed in 4 fetal lambs at 120 to 130 days' gestation. An artificial sinusoidal heart rate pattern was obtained by administration of atropine sulfate and arginine vasopressin in 9 tests. An autoregression model was used to compare the spectral patterns before and during the sinusoidal heart rate pattern. RESULTS: Marked decreases in low-frequency (0.025-0.125 cycles/beat) and high-frequency (0.2-0.5 cycles/beat) areas were observed in the presence of the sinusoidal heart rate pattern. However, there were no significant changes in the very-low-frequency area (0.01-0.025 cycles/beat), which corresponds to the frequency of the sinusoidal heart rate pattern. CONCLUSION: The sinusoidal heart rate pattern may represent a very low-frequency component inherent in fetal heart rate variability that appears when low- and high-frequency components are reduced as a result of strongly suppressed autonomic nervous activity. PMID- 10819864 TI - Neonatal alloimmune thrombocytopenia: antenatal management. AB - OBJECTIVE: The optimal management of pregnancies at risk for neonatal alloimmune thrombocytopenia is debated. Proposed management includes the administration of intravenous immunoglobulin and serial determination of the fetal platelet count. The aims of our study were to determine the effectiveness and likely mechanism of action of intravenous immunoglobulin and to evaluate the safety of cordocentesis in cases of neonatal alloimmune thrombocytopenia. STUDY DESIGN: Eighteen mother infant pairs were studied. All were at risk for neonatal alloimmune thrombocytopenia on the basis of delivery of a previously affected infant and confirmation of specific maternal antiplatelet antibodies. The pertinent antigen was HPA-1a in 13 cases, HPA-3a in 2 cases, and undetermined in 3 cases. Serial cordocenteses were used to determine fetal platelet counts. If the platelet count was <50,000/microL before 37 weeks' gestation, treatment was initiated with intravenous immunoglobulin administered to either the fetus (n = 2) or the mother (n = 8). In 3 cases fetal and maternal immunoglobulin G levels were determined before and after treatment. RESULTS: Seven (39%) fetuses had adequate platelet counts, were not treated, and were delivered of infants with normal platelet counts. Eleven (61%) fetuses were thrombocytopenic. Eight thrombocytopenic infants were treated with maternally administered intravenous immunoglobulin. In 6 (75%) of 8 cases the fetal platelet count increased after administration of intravenous immunoglobulin, but 2 fetuses remained severely thrombocytopenic. Two thrombocytopenic fetuses were treated with intravenous immunoglobulin infusion directly into the umbilical vein; both remained thrombocytopenic. Moreover, fetal immunoglobulin G levels did not correlate well with the response to intravenous immunoglobulin. Two (5.3%) of 38 cordocenteses were complicated by hemorrhagic complications, necessitating immediate cesarean delivery despite the use of prophylactic platelet transfusion in one case. CONCLUSION: Severe fetal alloimmune thrombocytopenia does not always occur in subsequent fetuses. Thus either fetal antigen status or platelet counts or both of these are necessary to determine whether treatment is needed. The effect of intravenous immunoglobulin on raising the fetal platelet count is inconsistent and appears to be caused by maternal or placental factors rather than a direct inhibition of fetal platelet destruction by immunoglobulin. The risk of hemorrhagic complications from cordocentesis in pregnancies complicated by neonatal alloimmune thrombocytopenia is higher than generally appreciated and is not always avoided by platelet transfusion at the time of the procedure. PMID- 10819865 TI - Effect of external cephalic version at term on fetal circulation. AB - OBJECTIVE: We sought to investigate the subclinical effect of external cephalic version on fetal circulation. STUDY DESIGN: A prospective observational study was conducted on 136 subjects who had external cephalic version at or beyond 36 weeks of gestation without clinical complication. Doppler ultrasonographic studies of the umbilical and middle cerebral circulations were performed before and after the external cephalic version. The following Doppler indexes were measured: (1) the pulsatility index of the umbilical artery, which represents disturbance of placental circulation, and (2) the pulsatility index of the fetal middle cerebral artery, which represents fetal response. The Wilcoxon signed rank test was used for all statistical analyses. RESULTS: There was no significant difference in pulsatility index of the umbilical artery before and after external cephalic version (P =.674). There was a statistically significant reduction in the pulsatility index of the middle cerebral artery after external cephalic version (P =.043), and this difference existed only among multiparous women (P =.029), among those in whom the external cephalic version was considered to be difficult (P =.038), and when the placenta was posteriorly located (P =.028). The reduction in pulsatility index was not related to whether the external cephalic version was successful. In all cases the Doppler indexes remained within the normal ranges, and there were no associated fetal complications. CONCLUSION: External cephalic version was not associated with any significant disturbance of placental resistance to blood flow. Conversely, external cephalic version was associated with a significant reduction in the pulsatility index of the middle cerebral circulation, especially among the multiparous women, after a difficult procedure or in those with a posterior placenta. This probably represents a normal fetal physiologic response to manipulation of the fetal head. PMID- 10819866 TI - Effect of single versus multiple courses of antenatal corticosteroids on maternal and neonatal outcome. AB - OBJECTIVE: Treatment of pregnant mothers with a single course of antenatal corticosteroids significantly reduces neonatal mortality and morbidity. Multiple weekly courses are often given. However, the safety and efficacy of repeated courses of antenatal corticosteroids have not been adequately studied. STUDY DESIGN: A retrospective study was performed for 609 mothers and their 713 infants who were treated with 1 to 12 courses of antenatal corticosteroids. Data for 369 singleton preterm infants born at < or =34 weeks' gestation, 210 multiple gestations, and 134 infants delivered at > or =35 weeks' gestation were analyzed separately. RESULTS: The incidence of respiratory distress syndrome was 45% for single-course and 35% for multiple-course groups (P =.005; odds ratio, 0.44; 95% confidence interval, 0.25-0.79). The multiple-course group also had significantly less patent ductus arteriosus (20% vs 13%; P =.016). Incidence of death before discharge and other neonatal morbidities were similar. The multiple-course group had a reduction of 0.46 +/- 0.19 cm in head circumference at birth (P =.013) when adjusted for gestational age and preeclampsia. The 2 groups had similar birth weights. Infants born at > or =35 weeks' gestation, multiple-gestation infants, and infants who were born >7 days after the last corticosteroid dose had similar outcomes, regardless of the number of courses they received. Mothers treated with multiple courses compared with a single course had a significantly higher incidence of postpartum endometritis (P =.013), even though they had a lower incidence of prolonged rupture of membranes (24% vs 33%, P =.06) and similar cesarean delivery rates. CONCLUSION: Exposure to multiple courses of antenatal corticosteroids compared with a single course resulted in a significant reduction in the incidence of respiratory distress syndrome in singleton preterm infants delivered within a week of the last corticosteroid dose. This was associated with a reduction in birth head circumference and an increased incidence of maternal endometritis. Whether the potential benefits of repeated therapy clearly outweigh the risks will ultimately be determined in randomized prospective controlled trials. PMID- 10819867 TI - Birth weight references for twins. AB - OBJECTIVE: We sought to formulate fetal and birth weight references for twins from longitudinal data. STUDY DESIGN: This historic cohort study was based on 1831 pregnancies of twins born alive at >/=28 weeks' gestation from Baltimore, Maryland; Miami, Florida; Charleston, South Carolina; and Ann Arbor, Michigan. RESULTS: When we compared singletons and twins, the percentiles of twins fell substantially below the 10th percentile of singletons by 28 weeks' gestation, below the 50th percentile by 30 weeks' gestation, and below the 90th percentile by 34 weeks' gestation. The difference at the 50th percentile was 147 g (10%) at 30 weeks' gestation, 242 g (14%) at 32 weeks' gestation, 347 g (17%) at 34 weeks' gestation, 450 g (19%) at 36 weeks' gestation, 579 g (22%) at 38 weeks' gestation, and 772 g (27%) at 40 weeks' gestation. CONCLUSION: This new reference demonstrates that, although the overall pattern of fetal growth is slower for twins versus singletons from about 30 weeks' gestation, well-grown twins and singletons do not differ as much as previously believed. PMID- 10819868 TI - A common mutation in the 5,10-methylenetetrahydrofolate reductase gene as a new risk factor for placental vasculopathy. AB - OBJECTIVE: This study was undertaken to investigate whether the cytosine-to thymine substitution at nucleotide 677 (C677T) in the 5, 10 methylenetetrahydrofolate reductase gene is a risk factor for placental vasculopathy (abruptio placentae or placental infarction with fetal growth restriction). STUDY DESIGN: This case-control study enrolled 165 women with placental vasculopathy and 139 matched control women with normal pregnancy outcomes. Measurements included fasting total plasma homocysteine concentration, serum and red blood cell folate concentrations, serum vitamin B(12) concentration, whole-blood vitamin B(6) concentration, and analysis of the 5, 10 methylenetetrahydrofolate reductase gene C677T mutation. RESULTS: The median total plasma homocysteine concentration was significantly higher in the study group than in the control group (P <.01; odds ratio >97.5th percentile, 4.66; 95% confidence interval, 1.55-14.0). Homozygous genotype for the mutated 5,10 methylenetetrahydrofolate reductase gene was found in 12% of the study group and 5% of the control group (odds ratio, 2.45; 95% confidence interval, 1.00-6.02). CONCLUSIONS: Mild hyperhomocysteinemia was confirmed among women with placental vasculopathy, for which homozygosity for a mutated 5, 10 methylenetetrahydrofolate reductase gene was found to be a new risk factor. The risk of placental vasculopathy probably increases in conditions of low serum folate concentration. PMID- 10819869 TI - Cornual heterotopic pregnancy: contemporary management options. AB - This review covers the clinical presentations, treatments, and outcomes of cornual heterotopic pregnancies reported in the literature. Infertile women with a history of ectopic pregnancy, tubal surgery, or disease are at increased risk for cornual heterotopic pregnancy when they undergo in vitro fertilization. Women who have undergone bilateral salpingectomy also seem to be predisposed to this condition when they undergo in vitro fertilization. We recommend that these patients be followed up closely after a successful in vitro fertilization cycle with monitoring of serum beta-human chorionic gonadotropin levels and serial transvaginal ultrasonography because of the high associated morbidity. Laparotomy remains the treatment of choice for rupture of a cornual heterotopic pregnancy. In the absence of cornual rupture, however, medical management is an option that eliminates the risk of surgery and anesthesia and results in outcomes similar to those associated with surgical treatment. Currently there is insufficient evidence to recommend any single treatment modality, and the decision should be based on such factors as clinical presentation, surgeon's expertise, side effects, overall cost, and the patient's preference. PMID- 10819870 TI - HELLP (hemolysis, elevated liver enzymes, and low platelets) needs help. PMID- 10819872 TI - Obstetric maneuvers in cases of shoulder dystocia. PMID- 10819896 TI - Methamphetamine decreases calcium-calmodulin dependent protein kinase II activity in discrete rat brain regions. AB - A Ca(2+)/calmodulin-dependent signaling cascade has been implicated in the regulation of dopaminergic neurotransmission after chronic administration of amphetamine and methamphetamine (METH). We found a decrease in Ca(2+)/calmodulin dependent protein kinase II (CaM-kinase II) activity in five regions of the rat brain (parietal cortex; frontal cortex; hippocampus; striatum; and nucleus accumbens) after a single injection of METH. Pretreatment with the selective dopamine D1 receptor antagonist SCH 23390 prevented the acute METH-induced decrease in CaM-kinase II activity in the parietal cortex, striatum, nucleus accumbens, and substantia nigra/ventral tegmental area (SN/VTA). Pretreatment with the N-methyl-D-aspartate receptor antagonist MK-801 significantly restored the acute METH-induced decrease in CaM-kinase II activity in the parietal cortex, nucleus accumbens, and SN/VTA. Striatal CaM-kinase II activity was still significantly lower than that of the chronic saline-treated controls after a 1 week, but not a 4-week, abstinence from chronic administration of METH. A METH challenge after a 4-week abstinence period induced a more pronounced decrease in CaM-kinase II activity in rats chronically injected with METH than in rats chronically injected with saline. Western blot analysis revealed that the amount of CaM-kinase II protein was not altered after a single METH injection or after chronic METH injections, compared with saline-treated controls. However, amounts of phosphorylated (Thr(286)) CaM-kinase II in the parietal cortex, striatum and SN/VTA were significantly decreased at 3 h after an acute METH injection compared with saline-treated controls. These results suggest that dephosphorylation of CaM kinase II may contribute to the decreased enzyme activities induced by acute METH administration, and that chronic treatment with METH leads to an enhanced capacity of METH to decrease CaM-kinase II activity after an extended withdrawal period. PMID- 10819897 TI - Expression of complexin I and II mRNAs and their regulation by antipsychotic drugs in the rat forebrain. AB - Complexin (cx) I and II are homologous synaptic protein genes which are differentially expressed in mouse and human brain and differentially affected in schizophrenia. We characterized the distribution of cx I and II mRNAs in rat forebrain and examined whether their abundance, or the transcript of the synaptic marker synaptophysin, is affected by 14 days' administration of antipsychotic drugs (haloperidol, chlorpromazine, risperidone, olanzapine, or clozapine). Cx I mRNA predominated in medial habenula, medial septum-diagonal band complex, and thalamus, whereas cx II mRNA was more abundant in most other regions, including isocortex and hippocampus. Within the hippocampus, cx I mRNA was primarily expressed by interneurons and cx II mRNA by granule cells and pyramidal neurons. Localized cx II mRNA signal was seen in the dentate gyrus molecular layer, suggestive of its transport into granule cell dendrites. Antipsychotic treatment produced selective, modest effects on cx mRNA expression. Cx I mRNA was elevated by olanzapine in dorsolateral striatum and frontoparietal cortex, while the abundance of cx II mRNA relative to cx I mRNA was decreased in both areas by olanzapine and haloperidol. Chlorpromazine increased cx II mRNA in frontoparietal cortex and synaptophysin mRNA in dorsolateral striatum. In summary, the data have implications both for understanding the effects of antipsychotic medication on synaptic organization, and for synaptic protein expression studies in patients treated with the drugs. PMID- 10819898 TI - beta-adrenergic receptors primarily are located on the dendrites of granule cells and interneurons but also are found on astrocytes and a few presynaptic profiles in the rat dentate gyrus. AB - In the rat dentate gyrus, beta-adrenergic receptor (beta-AR) activation is thought to be important in mediating the effects of norepinephrine (NE). beta-AR immunoreactivity (beta-AR-I) was localized in this study by light and electron microscopy in the rat dentate gyrus by using two previously characterized antibodies to the beta-AR. By light microscopy, dense beta-AR-I was observed in the somata of granule cells and a few hilar interneurons. Diffuse and slightly granular beta-AR-I was found in all laminae, although it was most noticeable in the molecular layer. Ultrastructurally, the cytoplasm of granule cell and interneuronal perikarya (some of which contained parvalbumin immunoreactivity) contained beta-AR-I. beta-AR-I was associated primarily with the endoplasmic reticula; however, a few patches were observed near the plasmalemma. Quantitative analysis revealed that the greatest proportion of beta-AR-labeled profiles was found in the molecular layer. The majority of beta-AR-labeled profiles were either dendritic or astrocytic. In dendritic profiles, beta-AR-I was prominent near postsynaptic densities in large dendrites, many of which originated from granule cell somata. Moreover, some beta-AR-I was found in dendritic spines, sometimes affiliated with the spine apparati. Astrocytic profiles with beta-AR-I were commonly found next to unlabeled terminals which formed asymmetric (excitatory-type) synapses with dendritic spines. Additionally, beta-AR-I was observed in a few unmyelinated axons and axon terminals, many of which formed synapses with dendritic spines. Dual-labeling studies revealed that axons and axon terminals containing tyrosine hydroxylase (TH), the catecholamine synthesizing enzyme, often were near both neuronal and glial profiles containing beta-AR-I. These studies demonstrate that hippocampal beta-AR-I is localized: 1) principally in postsynaptic sites on granule cells and a few interneurons (some of which were basket cells); and 2) in glial processes. These observations add further support to the contention that beta-AR-activation modulates synaptic function through disparate pathways: directly, at either postsynaptic densities or presynaptic processes, or indirectly, through adjacent glial processes. PMID- 10819899 TI - Substantia nigra glutamate antagonists produce contralateral turning and basal ganglia Fos expression: interactions with D1 and D2 dopamine receptor agonists. AB - Experiments measuring behavior and immediate-early gene expression in the basal ganglia can reveal interactions between dopamine (DA) and glutamate neurotransmission. Nigrostriatal DA projections influence two striatal efferent pathways that, in turn, directly and indirectly influence the activity of the substantia nigra pars reticulata (SNr). This report tests the interactions between striatal DA receptors and nigral glutamate receptors on basal ganglia function by examining both contralateral turning and Fos immunoreactivity in striatum and pallidum following unilateral intranigral microinfusions of glutamate antagonists given to intact and 6-OHDA-lesioned rats. The NMDA antagonist AP5 (1 microg), or the AMPA/kainate antagonist DNQX (0.015-1.5 microg), injected into the SNr (0.5 microl) elicited contralateral turning as well as both striatal and pallidal Fos expression. Moreover, intranigral DNQX elicited more turning and greater numbers of Fos-positive striatal neurons in 6 OHDA-lesioned animals than in unlesioned controls, suggesting that the 6-OHDA injection induces functional changes in nigral glutamate transmission. In 6-OHDA lesioned rats, systemic injections of the DA D1 receptor agonist SKF38393 (0.5 mg/kg, i.p.) increased striatal Fos expression due to intranigral DNQX. In contrast, the D2 agonist quinpirole (0.1 mg/kg, i.p.) decreased striatal Fos expression but increased the pallidal Fos arising from intranigral AP5. In additional experiments, both intact and 6-OHDA-lesioned rats were given simultaneous intranigral and intrastriatal infusions and turning and pallidal Fos expression were measured. 6-OHDA-lesioned rats given 5 microg of intrastriatal quinpirole exhibited both turning and pallidal Fos that was significantly increased by intranigral AP5. These results indicate that the opposing influences of D2 agonists and endogenous nigral glutamate transmission are mediated by striatal D2 receptors. Finally, the behavioral effects of intranigral glutamate antagonism can be dissociated from the effects on striatal or pallidal immediate early gene expression. PMID- 10819900 TI - Serotonin(2C) receptors tonically suppress the activity of mesocortical dopaminergic and adrenergic, but not serotonergic, pathways: a combined dialysis and electrophysiological analysis in the rat. AB - The present study evaluated, via a combined electrophysiological and dialysis approach, the potential influence of serotonin (5-HT)(2C) as compared to 5-HT(2A) and 5-HT(2B) receptors on dopaminergic, adrenergic, and serotonergic transmission in frontal cortex (FCX). Whereas the selective 5-HT(2A) antagonist MDL100,907 failed to modify extracellular levels of dopamine (DA), noradrenaline (NA) or 5 HT simultaneously quantified in single dialysate samples of freely-moving rats, the 5-HT(2B)/5-HT(2C) antagonist SB206,553 dose-dependently increased levels of DA and NA without affecting those of 5-HT. This action was attributable to 5 HT(2C) receptor blockade inasmuch as the selective 5-HT(2C) antagonist SB242,084 likewise increased FCX levels of DA and NA, whereas the selective 5-HT(2B) antagonist SB204,741 was ineffective. Further, the preferential 5-HT(2C) receptor agonist Ro60-0175 dose-dependently depressed FCX levels of DA. The suppressive influence of 5-HT(2C) receptors on DA release was also expressed on mesolimbic and nigrostriatal dopaminergic pathways, in that levels of DA in nucleus accumbens and striatum were likewise reduced by Ro60-0175 and elevated, though less markedly, by SB206,553. In line with the above findings, Ro60-0175 dose dependently decreased the firing rate of ventrotegmental dopaminergic and locus coeruleus (LC) adrenergic perikarya, whereas their activity was dose-dependently enhanced by SB206,553. Furthermore, SB206,553 transformed the firing pattern of ventrotegmental dopaminergic neurons into a burst mode. In contrast to SB206,553, MDL100,907 had little affect on the firing rate of dopaminergic or adrenergic neurons. In conclusion, as compared to 5-HT(2A) and 5-HT(2B) receptors, 5-HT(2C) receptors exert a tonic, suppressive influence on the activity of mesocortical - as well as mesolimbic and nigrostriatal - dopaminergic pathways, likely via indirect actions expressed at the level of their cell bodies. Frontocortical adrenergic, but not serotonergic, transmission is also tonically suppressed by 5 HT(2C) receptors. PMID- 10819901 TI - Presynaptic dopamine D(4) receptor localization in the rat nucleus accumbens shell. AB - Dopamine D(4) receptors in the nucleus accumbens shell (AcbSh) are thought to play a key role in mediating the locomotor and sensitizing affects of psychostimulants, as well as the therapeutic efficacy of atypical antipsychotic drugs. We used electron microscopic immunocytochemistry to determine the functional sites for endogenous and exogenous D(4) receptor activation in this region. Of 1,090 D(4) receptor-labeled profiles observed in the AcbSh of rat brain, 65% were axons and axon terminals, while 22% were dendrites and dendritic spines. Within axons and terminals, D(4) receptor immunoreactivity was localized to segments of the plasma membrane and membranes of nearby vesicles. The axon terminals were morphologically heterogenous, varying in size and content of either all small synaptic vesicles (ssv), or ssv and large dense-core vesicles. The labeled terminals occasionally formed asymmetric excitatory-type axospinous synapses, but the majority were without recognizable synaptic specializations. In a separate series of tissue sections that were processed for dual-labeling of the D(4) receptor and the catecholamine synthesizing enzyme, tyrosine hydroxylase (TH), 56% of all observed associations were appositions between differentially labeled axonal profiles, and 17% were terminals that contained immunoreactivity for both antigens. Dendritic spines containing D(4) receptor-labeling also received convergent input from TH-immunoreactive terminals and unlabeled terminals forming asymmetric synapses. These results provide the first ultrastructural evidence for a major presynaptic, and a more minor postsynaptic, involvement of D(4) receptors in dopaminergic modulation of excitatory transmission in the AcbSh. PMID- 10819902 TI - Basic fibroblast growth factor (bFGF) acts on both neurons and glia to mediate the neurotrophic effects of astrocytes on LHRH neurons in culture. AB - Luteinizing hormone-releasing hormone (LHRH) neurons play a pivotal role in the neuroendocrine control of mammalian reproduction. Astrocytes were shown to be involved in the regulation of LHRH neuronal function, but little is known about the contribution of astroglial-derived factors in the regulation of LHRH neuron development. In order to gain insight into the mechanisms regulating the development of these cells, at morphological and biochemical levels we characterized the neurotrophic effects exerted by young astrocytes (maintained in culture for 8 days in vitro) and old astrocytes (maintained 26 days) on the differentiation, proliferation, and phenotypic expression of immortalized hypothalamic LHRH (GT(1-1)) neurons in vitro. Culturing GT(1-1) cells in the presence of young glia for different time intervals caused a marked acceleration in the acquisition of their neuronal phenotype. At all times examined, GT(1-1) cells cocultured with young glia exhibited a significantly greater extension of processes/cell, larger number of processes/cell and greater surface area of growth cones than GT(1-1) cells grown over nonglial adhesive substrates (polylysine). By contrast, when GT(1-1) neurons were cocultured with old glia, the length of neuronal processes and the growth cone surface area were significantly lower than in control GT(1-1) neurons cultured in the absence of glia. At 3 days in vitro (DIV), GT(1-1) neurons cocultured with young glia exhibited a 50% lower incorporation of [(3)H]thymidine than GT(1-1) neurons cultured without glia. By contrast, in the presence of old glia [(3)H]thymidine incorporation was significantly higher in cells cocultured with glia than in GT(1 1) neurons cultured alone. Localization of the proliferating cells by dual immunohistochemical staining revealed that the incorporation of bromodeoxiuridine (BrdU) was restricted to nuclei of GT(1-1) neurons when these were cocultured with young glia, but associated with both neurons and astrocytes in the presence of old glia. At the functional level, coculture of GT(1-1) neurons with young glia increased the spontaneous release of LHRH as compared to GT(1-1) neurons grown in the absence of glia. By contrast, in the presence of old glia LHRH release in the medium was significantly lower than in controls. Conditioned medium of young glia (ACM-Y) induced significant neurotrophic and functional effects on GT(1-1) cells, but these effects were 50% less potent than the coculture itself. Heat denaturation of ACM-Y totally abolished its neurotrophic and functional properties, indicating that they involved a peptide factor. Suppression of bFGF activity in ACM-Y reduced its neurotrophic activity by approximately 40%, but did not affect its LHRH release-promoting effects. By contrast, neutralization of endogenous bFGF activity in GT(1-1) neurons cocultured with young glia counteracted both neurotrophic and functional effects of young glia. Treatment of old glia with bFGF rescued its neurotrophic and functional effects on GT(1-1) cells. Moreover, the ACM of aged bFGF-treated old glia was the most powerful neurotrophic stimulus for GT(1-1) neurons. These results suggest that: 1) soluble peptidic factors, including bFGF, and mechanism(s) requiring coculture are responsible for the highly potent neurotrophic and functional effects of young glia; 2) the inhibitory effects of old glia on neurite outgrowth and LHRH release are mediated in part by soluble inhibitory molecules and in part by factors requiring coculture with old glia; 3) old glia may revert to a growth-supporting state when treated with bFGF and this functional shift involves a diffusible molecule with potent neurotrophic and functional effects on immortalized LHRH neurons. (c) 2000 Wiley-Liss, Inc. PMID- 10819903 TI - Maturation of NK1 receptor involvement in the nociceptive response to formalin. AB - Administration of NK1 antagonists in adult animals attenuates the nociceptive response in the formalin test, indicating that the neurokinins and the NK1 receptor play a role in mediating this pain response. The number and distribution of NK1 receptors change dramatically during development, and the age at which they become involved in pain processing is not known. We examined the role of NK1 receptors in the formalin model in rats ranging in age between 3- and 21-days old. An NK1 antagonist, CP99,994, and its less active enantiomer CP100,263 were administered to the spinal cord (intrathecal), systemically (subcutaneous), or locally (intraplantar). Intrathecal administration of CP99,994, but not CP100,263, attenuated pain behaviors in the second phase of the formalin response in 14-day and 21-day old rats, but did not alter the pain response in 3-day or 10 day old rats. CP99,994 also reduced the expression of the c-fos protein in the superficial dorsal horn of 21-day old rats. Systemic and intraplantar injection of either CP99,994 or CP100,263 reduced the pain response to formalin in 3-day and 21-day old rats, suggesting a non-NK1 mediated mechanism of action. These results indicate that, within the spinal cord, NK1 receptors start to play a role in the pain response to formalin between 10 and 21 days. Moreover, analgesia induced by systemic or local injection of NK1 antagonists involves mechanisms other than, or in addition to, the NK1 receptor. PMID- 10819904 TI - Non-NMDA receptor-mediated mechanisms are involved in levodopa-induced motor response alterations in Parkinsonian rats. AB - Chronic dopaminomimetic administration to parkinsonian animal models or Parkinson's disease patients leads to characteristic alteration in motor response. Previous studies suggested that the nonphysiologic stimulation of dopaminergic receptors on striatal medium spiny neurons enhances the synaptic efficacy of juxtaposed glutamate receptors of the N-methyl-D-aspartate (NMDA) subtype. Resultant NMDA receptor sensitization due to differential changes in subunit phosphorylation appears to favor alterations in striatal output in ways that influence motor function. To detail the involvement of NMDA receptors further as well as to determine whether similar functional changes might develop in alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) receptors, the effects of selective antagonist of AMPA receptors (6-nitro-7-sulfamoyl-benzo[f] quinoxaline-2,3 (1H,4H)-dione sodium salt, NBQX, 10 mg/kg) on levodopa-induced response alterations in 6-hydroxydopamine (6-OHDA) lesioned rats were compared with drugs which act competitively (3-(+/-)-2-carboxypiperazin-4-yl)-propyl-1 phosphonicacid, CPP, 6.25 mg/kg) or noncompetitively (dextromethorphan, 40 mg/kg) to block NMDA receptors, or a nonselective inhibitor of glutamatergic transmission (2-amino-6-trifluoromethoxy benzothiazole, riluzole, 5 mg/kg). We found that the shortened duration of the motor response to levodopa, which underlies human wearing-off fluctuations, was reversed to a similar degree by the acute coadministration of CPP, NBQX, or riluzole (n = 4-6) but dextromethorphan did not. These observations strengthen the possibility that a reduction in levodopa-associated changes in motor response by inhibitors of glutamatergic transmission acting generally or selectively at the glutamate binding-sites may relate to their ability to attenuate pathologic gain in striatal glutamatergic function. The capacity of NBQX to reverse these altered responses suggests that an enhanced synaptic efficacy of striatal AMPA receptors may also participate in the generation of these motor response changes in levodopa-treated parkinsonian rats. PMID- 10819905 TI - Direct effects of 3,4-methylenedioxymethamphetamine (MDMA) on serotonin or dopamine release and uptake in the caudate putamen, nucleus accumbens, substantia nigra pars reticulata, and the dorsal raphe nucleus slices. AB - We examined the effects of pressure ejected 3, 4-methylenedioxymethamphetamine (MDMA) from a micropipette on direct chemically stimulated release, and on electrically stimulated serotonin (5-HT) or dopamine (DA) release in the caudate putamen (CPu), nucleus accumbens (NAc), substantia nigra pars reticulata (SNr), and the dorsal raphe nucleus (DRN) brain slices of rat, using fast cyclic voltammetry (FCV). MDMA is electroactive, oxidising at +1100 mV. When the anodic input waveform was reduced from +1.4 to +1.0 volt, MDMA was not electroactive. Using this waveform, pressure ejection of MDMA did not release 5-HT or DA in brain slices prepared from any of the nuclei studied. MDMA significantly potentiated electrically stimulated 5-HT release in the SNr and DA release in CPu. In the DRN or in the NAc, MDMA was without effect on peak electrically stimulated 5-HT or DA release. The rates of neurotransmitter uptake, expressed as t(1/2), were in all cases significantly decreased after MDMA. The results indicate that MDMA, unlike (+)amphetamine, is not as a releaser of DA or 5-HT, it is a potent inhibitor of both DA and 5-HT uptake. PMID- 10819906 TI - Sequential changes in the synaptic structural profile following long-term potentiation in the rat dentate gyrus. II. Induction/early maintenance phase. AB - Long-term potentiation (LTP), one of the most compelling models of learning and memory, has been associated with changes in synaptic morphology. In this study, LTP was induced and animals were sacrificed 1 h after the stimulation of the LTP group (induction / early maintenance phase). Synapses in the directly stimulated middle third of the dentate gyrus molecular layer (MML) were examined while synapses from the inner third of the dentate molecular layer (IML) of the LTP animals and both the MML and the IML of implanted animals served as controls. The total number of synapses per neuron, synaptic curvature, the presence of synaptic perforations, and the maximum length of the synaptic contact and active zone were examined. No overall change in the number of synapses per neuron was observed in the LTP tissue. LTP was associated with a significant increase in the proportion of perforated and irregular-shaped synapses compared to controls. The increase in perforated synapses was particularly apparent in the proportion of concave perforated synapses. Nonperforated concave synapses were found to be significantly larger in potentiated tissue. The total synaptic length per neuron of synapses in a concave configuration was also significantly higher following potentiation. These results suggest that the specific structural profile associated with 1-h post-LTP induction, which differed from the profile observed at 24 h post-induction, may represent a unique early phase of synaptic remodeling in a series of changes observed during LTP induction, maintenance, and decay. PMID- 10819907 TI - Dynamic dopamine receptor interactions in the core and shell of nucleus accumbens differentially coordinate the expression of unconditioned motor behaviors. AB - Many neurochemical and behavioral functions mediated by dopamine require the dynamic interaction between dopamine receptors. We examined the behavioral effects evoked by microinjections of drugs with relative selectivity for specific dopamine receptors into the nucleus accumbens (Acb). The results showed that, at behaviorally inactive doses, the dopamine D1-class receptor agonist SKF 38393 switched the behavioral profile induced by injections of the dopamine D2-class receptor agonist quinpirole into the Acb, from sedation, yawning, and motor inhibition to hyperactive-like behavior. Further, the effects of injections of the dopamine D2-class receptor agonist (+)-PD 128907 into the shell of Acb, including suppression of rearing, locomotion, and grooming, and induction of oral dyskinesia, yawning, and sedation, could not be distinguished from those elicited by (+)-PD 128907 following infusions into the core of Acb. However, the behavioral effects elicited by coadministration of SKF 38393 and (+)-PD 128907 into the core or the shell of Acb showed a striking anatomical specificity. The infusion of SKF 38393 plus (+)-PD 128907 into the core, but not into the shell, of Acb modified the pattern of responses induced by (+)-PD 128907, inducing behavioral hyperactivity. These results suggest critical differences in the functional interaction between dopamine receptors in the core and the shell of the Acb and reveal a mechanism of behavioral switching in the core of Acb by virtue of which dopamine D1-class receptors regulate the transition from states of behavioral suppression to states of heightened psychomotor arousal. PMID- 10819908 TI - Nicotinic modulation of [(3)H]D-aspartate outflow from cultured cerebellar granule cells. AB - The effect of nicotine on basal and electrically evoked (20 Hz for 20 sec) [(3)H]D-aspartate efflux (assumed as an index of transmitter release) was studied in rat cerebellar granule primary cultures. Nicotine (10-100 nM) increased the basal efflux two to three times and concentration-dependently enhanced the electrically evoked efflux up to ten times. Higher drug concentration (1 microM) underwent rapid desensitization. Facilitation of the efflux was similarly reduced by the nicotinic acetylcholine receptor antagonists, alpha-bungarotoxin and mecamylamine, suggesting the involvement of at least two receptor subtypes containing and lacking alpha(7) subunits, respectively. Since the increased efflux induced by nicotine in granule cells kept at rest or depolarized by KCl 15 mM was antagonized by tetrodotoxin, the involvement of sodium channels by receptors located at preterminal sites was suggested. Taken together, these findings emphasize the role of the cholinergic input in granule cell function and in glutamatergic signaling. PMID- 10819909 TI - Nicotine-induced Fos expression in the nucleus accumbens and the medial prefrontal cortex of the rat: role of nicotinic and NMDA receptors in the ventral tegmental area. AB - We have previously shown that the nicotine-induced dopamine release in the nucleus accumbens can be attenuated by local administration into the ventral tegmental area (VTA), of antagonists at nicotinic and N-methyl-D-aspartate (NMDA), but not alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) receptors. In the present study, we investigated the role of nicotinic and NMDA receptors in the VTA for the expression of Fos-like immunoreactivity (FLI) in the shell and core of the nucleus accumbens and in the medial prefrontal cortex (mPFC) of the rat after acute nicotine administration. Systemically administered nicotine increased FLI in both the mPFC and the nucleus accumbens when compared to saline controls, although this effect was more pronounced, and reached statistical significance in the nucleus accumbens, especially in the core region. When mecamylamine was delivered by reverse dialysis into the VTA, the systemic nicotine-induced FLI was significantly attenuated in the nucleus accumbens. Similarly, the NMDA receptor antagonist 2-amino-5-phosphonopentanoic acid (AP-5), infused locally in the VTA, also antagonized the nicotine-induced FLI in the nucleus accumbens. Neither mecamylamine nor AP-5 alone affected basal FLI levels in any of the structures studied. Local administration of nicotine in the VTA increased FLI in the nucleus accumbens but not in the mPFC. Since the nicotine induced FLI is probably due to an increased dopamine release in both the nucleus accumbens and the mPFC, we conclude that FLI in the nucleus accumbens is mediated, to a large extent, through the activation of dopamine neurons via nicotinic and NMDA receptors in the VTA, whereas the nicotine-induced FLI in the mPFC is subjected to a differential control mechanism, tentatively involving nicotinic receptors at the terminal level of the mPFC-projecting dopamine neurons. PMID- 10819910 TI - Dopamine D(2) receptor quantification in extrastriatal brain regions using [(123)I]epidepride with bolus/infusion. AB - The iodinated benzamide epidepride, which shows a picomolar affinity binding to dopamine D(2) receptors, has been designed for in vivo studies using SPECT. The aim of the present study was to apply a steady-state condition by the bolus/infusion approach with [(123)I]epidepride for the quantification of striatal and extrastriatal dopamine D(2) receptors in humans. In this way the distribution volume of the tracer can be determined from a single SPECT image and one blood sample. Based on bolus experiments, an algorithm using conventional convolution arguments for prediction of the outcome of a bolus/infusion (B/I) experiment was applied. It was predicted that a B/I protocol with infusion of one third of the initial bolus per hour would be appropriate. Steady-state conditions were attained in extrastriatal regions within 3-4 h but the infusion continued up to 7 h in order to minimize the significance of individual differences in plasma clearance and binding parameters. A steady-state condition, however, could not be attained in striatal brain regions using a B/I protocol of 20 h, even after 11 h. Under near steady-state conditions a striatal:cerebellar ratio of 23 was demonstrated. Epidepride has a unique signal-to-noise ratio compared to [(123)I]IBZM but present difficulties for steady-state measurements of striatal regions. The bolus/infusion approach is particularly feasible for quantification of the binding potential in extrastriatal regions. PMID- 10819911 TI - Pindolol occupancy of 5-HT(1A) receptors measured in vivo using small animal positron emission tomography with carbon-11 labeled WAY 100635. AB - Positron emission tomography (PET), following an intravenous injection of [carbonyl-(11)C]WAY 100635, was used to image central 5-HT(1A) receptors in rat following pretreatment with graded doses of (-)-pindolol (0.001-3 mg/kg, i.v.). The use of PET had advantages over ex vivo radioligand binding methods in that it produced parametric image volumes and reduced errors due to inter-rat variability. Time-radioactivity curves from regions of interest (ROI) acquired from individual rats enabled the estimation of specific binding of the radioligand using a compartmental model with reference tissue input. Binding potential (BP) of [(11)C]WAY 100635 was estimated for frontal cortex and hippocampus (postsynaptic), and midbrain raphe nuclei (presynaptic). In the latter ROI, pindolol dose-dependently decreased BP. The saturation curve could be fitted to a single-site model up to the lowest dose of pindolol used, giving an ED(50) (dose to cause 50% occupancy) value of 0.26 +/- 0. 05 mg/kg, and inclusion of control (nonpindolol-treated) rats did not affect the fit. In contrast, in cortex and hippocampus ROI, low doses of pindolol caused an increase in BP compared with controls. Pindolol doses greater than approximately 0.1 mg/kg, resulted in a dose-dependent decrease in BP, and ED(50) values in cortex and hippocampus were estimated as 0.44 +/- 0.13 and 0.48 +/- 0.12 mg/kg, respectively. The increase in [(11)C]WAY 100635 binding at low pindolol doses is feasibly related to a decrease in basal receptor occupancy following reduced release of endogenous 5-HT. Considering the apparently greater potency of pindolol at the midbrain raphe ROI, this effect could be mediated via agonist activity at the autoreceptor. PMID- 10819912 TI - Altered responsiveness of medial prefrontal cortex neurons to glutamate and dopamine after withdrawal from repeated amphetamine treatment. AB - Using in vivo extracellular single-cell recording and microiontophoresis, we compared the responsiveness of medial prefrontal cortex (mPFC) neurons (layers V VI) to dopamine and glutamate in rats that had received repeated amphetamine or saline injections. Neurons from amphetamine-pretreated rats showed increased responsiveness to glutamate and decreased responsiveness to dopamine after three days of withdrawal, suggesting that the mPFC is transiently hyperexcitable during amphetamine withdrawal. Published 2000 Wiley-Liss, Inc. PMID- 10819913 TI - Dedicated extremity MR imaging of the knee: how low can you go? AB - Low-field strength, dedicated extremity MR imaging systems can provide clinically effective MR images at substantially lower costs, compared with high-field strength, whole-body MR imaging machines. Dedicated extremity units also reduce risks of patient safety and claustrophobia. The diagnostic performance of presently existing extremity MR scanners rivals that of high-field strength units. Focusing on the imaging of the knee joint, this article discusses issues of technique, imaging protocol, clinical roles, and economics, and reviews diagnostic efficacy studies. PMID- 10819914 TI - Current concepts and controversies in meniscal imaging. AB - Analyzing meniscal abnormalities on MR images depends on a thorough understanding of the normal appearance of meniscal structures and on strict adherence to the MR imaging criteria for meniscal tears. When proper MR imaging technique is followed, meniscal tears can be diagnosed more acutely than with clinical examination alone, even in the presence of associated ligament injuries. The value of MR imaging lies in its ability to reveal details of meniscal tears, including location, morphology, length, depth, and possible stability. If this MR imaging information can be synthesized, it will dramatically affect treatment planning by helping to predict meniscal tears that are potentially reparable and tears that might not need surgical intervention. PMID- 10819915 TI - Second time around: evaluating the postoperative anterior cruciate ligament. AB - Anterior cruciate ligament (ACL) injuries are especially common in the younger athletic population. In 1998, more than 100,000 ACL reconstructions were performed. MR imaging examination has proved to be valuable in imaging and evaluating ACL reconstructions. This article reviews different surgical reconstruction procedures, indications for postoperative MR imaging, and the MR imaging appearance seen in routine ACL reconstructions and in complications associated with ACL reconstructions. PMID- 10819916 TI - "Slip sliding Away": patellofemoral dislocation and tracking. AB - This article reviews the anatomy of the medial retinacular complex and the relationship of the anatomy to patellar dislocation. Clinical and radiographic findings of patellar dislocation are discussed, and the technical considerations and clinical usefulness of patellar tracking studies are addressed. PMID- 10819917 TI - Too much of a good thing: overuse injuries of the knee. AB - Overuse injuries constitute a distinct group of abnormalities arising from excessive stress, repeated microtrauma, inadequate injury repair, or pathologic motion of the knee. Chronic pain resulting from overuse injuries may progress to the point that the patient can no longer perform the inciting activity. MR image appearances of the more common overuse syndromes involving the soft tissues surrounding the knee are presented, pertinent anatomic features are reviewed, and the role of MR imaging in patient management in these disorders is discussed. PMID- 10819918 TI - "Dem bones": osteochondral injuries of the knee. AB - MR imaging plays a valuable role in the diagnosis and staging of osteochondral injuries of the femorotibial joint. Bone contusions may be the source of a patient's pain, and MR imaging characteristics of certain types may help to predict which contusions might progress to more serious osteochondral lesions. MR imaging also is vital in the diagnosis of occult osteochondral fractures and in accurately classifying displaced intra-articular fractures. Although osteochondral dissecans usually is diagnosed radiographically, MR imaging is the best noninvasive test for determining if an osteochondral fragment is unstable. Unstable lesions are a treatable cause of knee pain. PMID- 10819919 TI - Water on the knee: cysts, bursae, and recesses. AB - Many synovial-lined cavities occur around the knee, and MR imaging is capable of demonstrating the precise anatomic relationships of these spaces. Knowledge of normal anatomy, however, is necessary to accurately assess abnormal, fluid-filled structures. This article reviews the MR imaging appearance of normal and pathologic bursae and joint recesses, and the appearance of common cystic masses. Distinguishing features of several noncystic lesions occurring around the knee also are discussed because these can mimic bursae, recesses, and cysts. PMID- 10819920 TI - Loose fragments and other debris: miscellaneous synovial and marrow disorders. AB - Conventional MR imaging evaluation of the knee focuses mainly on the effects of acute trauma and degenerative conditions. Numerous other conditions that affect the knee may mimic clinical characteristics of common traumatic and degenerative disorders. Synovial and marrow abnormalities are frequently encountered and often unsuspected on routine MR imaging studies of the knee. This article presents the MR imaging appearance of the normal synovium, bone marrow, and commonly encountered abnormalities of these structures. PMID- 10819922 TI - A piece of My mind: who cares about tarawa? PMID- 10819921 TI - Scratching the surface: articular cartilage disorders in the knee. AB - Powerful techniques with MR imaging are being developed for evaluating articular cartilage. Pharmaceutical and biotechnology development is driven largely by the search for novel therapeutic solutions to the growing problem of arthritis in our aging society. Accordingly, imaging tools that will be used to direct patients to specific therapies and then to monitor treatment effectiveness and safety presently are being refined and validated in rigorous multicenter and multinational clinical trials aimed at gaining regulatory approval. As trials approach completion, radiologists can anticipate an increased demand for expertise and experience in evaluating articular cartilage disorders. This article reviews this novel mechanism for evolving imaging techniques for clinical use through clinical drug trials, lists the most promising MR imaging markers available for evaluating cartilage integrity, and speculates on how these techniques will find their way into clinical practice. PMID- 10819923 TI - Ambulatory gynecology. PMID- 10819925 TI - JAMA 100 years ago: DOES NOT TAKE A JOKE PMID- 10819924 TI - JAMA 100 years ago: ATTEMPTED CONTROL OF SECTION ON MATERIA MEDICA PMID- 10819926 TI - JAMA 100 years ago: EPILEPSY, RICKETS, AND THE LYMPHATIC CONSTITUTION PMID- 10819927 TI - National Health Service Corps faces reauthorization during a risky time. PMID- 10819928 TI - JAMA Web site enters new phase. PMID- 10819929 TI - Can male hormones really help women? PMID- 10819931 TI - Quick uptakes: resistant salmonella PMID- 10819930 TI - Addiction medicine specialists add a new therapeutic approach. PMID- 10819933 TI - Quick uptakes: fighting MS fatigue PMID- 10819932 TI - Quick uptakes: cancer recurrence predicted PMID- 10819934 TI - Quick uptakes: chemotherapy breath test PMID- 10819936 TI - AIDS-Related opportunistic illness and potent antiretroviral therapy PMID- 10819935 TI - AIDS-Related opportunistic illness and potent antiretroviral therapy. PMID- 10819937 TI - A critical pathway for treatment of community-acquired pneumonia. PMID- 10819938 TI - A critical pathway for treatment of community-acquired pneumonia PMID- 10819939 TI - Gifts to physicians from the pharmaceutical industry. PMID- 10819940 TI - Gifts to physicians from the pharmaceutical industry. PMID- 10819941 TI - Gifts to physicians from the pharmaceutical industry. PMID- 10819943 TI - Gifts to physicians from the pharmaceutical industry PMID- 10819942 TI - Gifts to physicians from the pharmaceutical industry. PMID- 10819944 TI - Gifts to physicians from the pharmaceutical industry PMID- 10819945 TI - William Osler at 150. PMID- 10819946 TI - William osler at 150 PMID- 10819947 TI - Arsenic contamination of museum artifacts repatriated to a Native American tribe. PMID- 10819948 TI - Causes of maternal mortality in Japan. AB - CONTEXT: Japan's maternal mortality rate is higher than that of other developed countries. OBJECTIVES: To identify causes of maternal mortality in Japan, examine attributes of treating facilities associated with maternal mortality, and assess the preventability of such deaths. DESIGN AND SETTING: Cross-sectional study of maternal deaths occurring in Japan between January 1, 1991, and December 31, 1992. SUBJECTS: Of 230 women who died while pregnant or within 42 days of being pregnant, 197 died in a hospital and had medical records available, 22 died outside of a medical facility, and 11 did not have records available. MAIN OUTCOME MEASURES: Maternal mortality rates per 100,000 live births by cause (identified by death certificate review and information from treating physicians or coroners); resources and staffing patterns of facilities where deaths occurred; and preventability of death, as determined by a 42-member panel of medical specialists. RESULTS: Overall maternal mortality was 9.5 per 100,000 births. Hemorrhage was the most common cause of death, occurring in 86 (39%) of 219 women. Seventy-two (37%) of 197 deaths occurring in facilities were deemed preventable and another 32 (16%) possibly preventable. Among deaths that occurred in a medical facility with an obstetrician on duty, the highest rate of preventable deaths (4.09/100,000 live births) occurred in facilities with 1 obstetrician. Among the 72 preventable deaths, 49 were attributed to 1 physician functioning as the obstetrician and anesthetist. While the unpreventable maternal death rate was highest in referral facilities, the preventable maternal death rate was 14 times lower in referral facilities than in transferring facilities. CONCLUSIONS: Inadequate obstetric services are associated with maternal mortality in Japan. Reducing single-obstetrician only delivery patterns and establishing regional 24-hour inpatient obstetrics facilities for high-risk cases may reduce maternal mortality in Japan. JAMA. 2000;283:2661-2667. PMID- 10819949 TI - Laboratory-based surveillance of Salmonella serotype Typhi infections in the United States: antimicrobial resistance on the rise. AB - CONTEXT: Multidrug-resistant Salmonella serotype Typhi infections have been reported worldwide, but data on the incidence of resistant strains in the United States are lacking. OBJECTIVES: To determine the incidence of antimicrobial resistant Salmonella Typhi infections and to identify risk factors for infection. DESIGN: Cross-sectional laboratory-based surveillance study. SETTING AND PARTICIPANTS: A total of 293 persons with symptomatic typhoid fever who had Salmonella Typhi isolates and epidemiological information submitted to US public health departments and laboratories from June 1, 1996, to May 31, 1997. MAIN OUTCOME MEASURES: Proportion of Salmonella Typhi isolates demonstrating resistance to 12 antimicrobial agents; patient epidemiological factors associated with drug-resistant infections. RESULTS: Median age was 21 years (range, 3 months to 84 years); 56% were male. Two hundred twenty-eight (80%) were hospitalized; 2 died. In the 6 weeks before illness onset, 81% of patients had traveled abroad. Seventy-four Salmonella Typhi isolates (25%) were resistant to 1 or more antimicrobial agent, and 51 (17%) were resistant to 5 or more agents, including ampicillin, chloramphenicol, and trimethoprim-sulfamethoxazole (multidrug resistant Salmonella Typhi [MDRST]). Although no resistance to ciprofloxacin or ceftriaxone was observed, 20 isolates (7%) were nalidixic acid-resistant (NARST). Patients with MDRST and NARST infections were more likely to report travel outside the United States, particularly to the Indian subcontinent (Bangladesh, India, and Pakistan) (odds ratio [OR], 29.3; 95% confidence interval [CI], 6.8 126.7; P<.001 and OR, 35.9; 95% CI, 3.4-377.3; P<.001, respectively). CONCLUSIONS: Our data suggest that ciprofloxacin and ceftriaxone are appropriate empirical therapy for suspected typhoid fever; however, resistance may be anticipated. Continued monitoring of antimicrobial resistance among Salmonella Typhi strains will help determine vaccination and treatment policies. JAMA. 2000;283:2668-2673. PMID- 10819950 TI - Association of coffee and caffeine intake with the risk of Parkinson disease. AB - CONTEXT: The projected expansion in the next several decades of the elderly population at highest risk for Parkinson disease (PD) makes identification of factors that promote or prevent the disease an important goal. OBJECTIVE: To explore the association of coffee and dietary caffeine intake with risk of PD. DESIGN, SETTING, AND PARTICIPANTS: Data were analyzed from 30 years of follow-up of 8004 Japanese-American men (aged 45-68 years) enrolled in the prospective longitudinal Honolulu Heart Program between 1965 and 1968. MAIN OUTCOME MEASURE: Incident PD, by amount of coffee intake (measured at study enrollment and 6-year follow-up) and by total dietary caffeine intake (measured at enrollment). RESULTS: During follow-up, 102 men were identified as having PD. Age-adjusted incidence of PD declined consistently with increased amounts of coffee intake, from 10.4 per 10,000 person-years in men who drank no coffee to 1.9 per 10,000 person-years in men who drank at least 28 oz/d (P<.001 for trend). Similar relationships were observed with total caffeine intake (P<.001 for trend) and caffeine from non-coffee sources (P=.03 for trend). Consumption of increasing amounts of coffee was also associated with lower risk of PD in men who were never, past, and current smokers at baseline (P=.049, P=.22, and P=.02, respectively, for trend). Other nutrients in coffee, including niacin, were unrelated to PD incidence. The relationship between caffeine and PD was unaltered by intake of milk and sugar. CONCLUSIONS: Our findings indicate that higher coffee and caffeine intake is associated with a significantly lower incidence of PD. This effect appears to be independent of smoking. The data suggest that the mechanism is related to caffeine intake and not to other nutrients contained in coffee. JAMA. 2000;283:2674-2679. PMID- 10819951 TI - Impact of a diagnostic cerebrospinal fluid enterovirus polymerase chain reaction test on patient management. AB - CONTEXT: Enterovirus (EV) infection, the most common cause of aseptic meningitis, can be rapidly diagnosed with an EV-specific reverse transcriptase polymerase chain reaction (EV-PCR) test. However, no studies have examined EV-PCR in a clinical context in which it is routinely used. OBJECTIVE: To determine the impact of EV-PCR testing on diagnosis and clinical management of suspected aseptic meningitis cases. DESIGN AND SETTING: Retrospective review of electronic medical records from a 220-bed tertiary care pediatric medical center in San Diego, Calif. PATIENTS: A total of 276 pediatric patients for whom a diagnostic EV-PCR test was performed during the calendar year 1998. MAIN OUTCOME MEASURES: Clinical parameters such as length of stay, medication use, and ancillary test use. RESULTS: One hundred thirty-seven patients (49.6%) had a positive cerebrospinal fluid EV-PCR result. Enterovirus-positive patients with results available before hospital discharge (n=95) had significantly fewer ancillary tests performed (26% vs 72% with at least 1 test performed; P<.001), received intravenous antibiotics for less time (median, 2.0 vs 3.5 days; P<.001), and had shorter hospital stays (median, 42 vs 71.5 hours; P<.001) than EV-negative patients (n=92). A positive EV-PCR result was associated with more rapid hospital discharge (median EV-PCR-to-discharge time, 5.2 hours) compared with a negative result (median EV-PCR-to-discharge time, 27.4 hours; P<.001). CONCLUSIONS: Our results suggest that a positive EV-PCR result may affect clinical decision making and can promote rapid discharge of patients, and that unnecessary diagnostic and therapeutic interventions can be reduced by use of EV-PCR testing. JAMA. 2000;283:2680-2685. PMID- 10819952 TI - Mortality and prehospital thrombolysis for acute myocardial infarction: A meta analysis. AB - CONTEXT: Early administration of thrombolysis for acute myocardial infarction (AMI) may improve survival if safely and appropriately delivered. No systematic reviews that have comprehensively examined this topic exist in the literature. OBJECTIVE: To perform a meta-analysis of randomized controlled trials of prehospital vs in-hospital thrombolysis for AMI measuring in-hospital mortality. DATA SOURCES: The Cochrane search strategy was used to search MEDLINE, EMBASE, and the Science Citation Index (1982-1999); Dissertation Abstracts (1987-1999); and Current Contents (1994-1999) for the terms thrombolysis, thrombolysis therapy, prehospital, and acute myocardial infarction. In addition, text and journal article bibliographies were hand searched, the National Institutes of Health Web site was reviewed, and primary authors and thrombolytic drug manufacturers were contacted for unpublished studies. STUDY SELECTION: Randomized controlled trials of prehospital vs in-hospital thrombolysis for AMI measuring all-cause hospital mortality were included. Two authors independently reviewed 175 citations by title, abstract, or complete article. After exclusion of 30 duplicate citations, 145 studies remained, of which 6 studies and 3 follow-up studies met the inclusion criteria. DATA EXTRACTION: Independent data abstraction by 2 reviewers blinded to the journal, title, and author was confirmed by consensus. Trial quality was independently assessed by 2 other coauthors, blinded to the author, title, journal, introduction, and discussion. DATA SYNTHESIS: The results of the 6 randomized trials (n=6434) were pooled and indicated significantly decreased all-cause hospital mortality among patients treated with prehospital thrombolysis compared with in-hospital thrombolysis (odds ratio, 0.83; 95% confidence interval, 0.70-0.98). Results were similar regardless of trial quality or training and experience of the provider. Estimated (SE) time to thrombolysis was 104 (7) minutes for the prehospital group and 162 (16) minutes for the in-hospital thrombolysis group (P=.007). CONCLUSIONS: Our meta-analysis suggests that prehospital thrombolysis for AMI significantly decreases the time to thrombolysis and all-cause hospital mortality. JAMA. 2000;283:2686-2692. PMID- 10819953 TI - A 52-year-old suicidal man. PMID- 10819954 TI - A 26-year-old woman seeking an abortion, 1 year later. PMID- 10819955 TI - What makes clinical research ethical? AB - Many believe that informed consent makes clinical research ethical. However, informed consent is neither necessary nor sufficient for ethical clinical research. Drawing on the basic philosophies underlying major codes, declarations, and other documents relevant to research with human subjects, we propose 7 requirements that systematically elucidate a coherent framework for evaluating the ethics of clinical research studies: (1) value-enhancements of health or knowledge must be derived from the research; (2) scientific validity-the research must be methodologically rigorous; (3) fair subject selection-scientific objectives, not vulnerability or privilege, and the potential for and distribution of risks and benefits, should determine communities selected as study sites and the inclusion criteria for individual subjects; (4) favorable risk-benefit ratio-within the context of standard clinical practice and the research protocol, risks must be minimized, potential benefits enhanced, and the potential benefits to individuals and knowledge gained for society must outweigh the risks; (5) independent review-unaffiliated individuals must review the research and approve, amend, or terminate it; (6) informed consent-individuals should be informed about the research and provide their voluntary consent; and (7) respect for enrolled subjects-subjects should have their privacy protected, the opportunity to withdraw, and their well-being monitored. Fulfilling all 7 requirements is necessary and sufficient to make clinical research ethical. These requirements are universal, although they must be adapted to the health, economic, cultural, and technological conditions in which clinical research is conducted. JAMA. 2000;283:2701-2711. PMID- 10819957 TI - Women's health: A call for papers PMID- 10819956 TI - Where should women deliver babies in Japan? PMID- 10819958 TI - Violence in G-rated animated films. AB - CONTEXT: Children's exposure to violence in the media is a possible source of public health concern; however, violence in children's animated films has not been quantified. OBJECTIVE: To quantify and characterize violence in G-rated animated feature films. DESIGN: Violence content was reviewed for all 74 G-rated animated feature films released in theaters between 1937 and 1999, recorded in English, and available for review on videocassette in the United States before September 1999. MAIN OUTCOME MEASURES: Duration of violent scenes, type of characters participating in violent acts (good, neutral, or bad), number of injuries/fatalities, and types of weapons used for each film. RESULTS: All 74 films reviewed contained at least 1 act of violence (mean duration, 9.5 minutes per film; range, 6 seconds-24 minutes). Analysis of time trends showed a statistically significant increase in the duration of violence in the films with time (P=.001). The study found a total of 125 injuries (including 62 fatal injuries) in 46 (62%) of the films. Characters portrayed as "bad" were much more likely to die of an injury than other characters (odds ratio, 23.2; 95% confidence interval, 8.5-63.4). A majority of the violence (55%) was associated with good or neutral characters dueling with bad characters (ie, using violence as a means of reaching resolution of conflict), and characters used a wide range of weapons in violent acts. CONCLUSIONS: Our content analysis suggests that a significant amount of violence exists in animated G-rated feature films. Physicians and parents should not overlook videocassettes as a source of exposure to violence for children. JAMA. 2000;283:2716-2720. PMID- 10819959 TI - Pains, strains, and fasciculations: lower extremity muscle disorders. AB - Muscle injuries that were suspected only on the basis of clinical examination are now readily demonstrated and characterized using MR imaging. Imaging is used not only for assessment of location, type, and severity of muscle injuries; it plays an important role in treatment planning, and the detection of complications. MR imaging also is useful in nontraumatic muscle disorders including congenital, inflammatory, and infectious myositis, and in cases of denervation, MR imaging is valuable for diagnosis, staging, and prognostication. In cases where the origin of myopathy is unknown, MR imaging can direct biopsy to increase diagnostic yield. PMID- 10819961 TI - Lysine-73 is involved in the acylation and deacylation of beta-lactamase. AB - Lysine 73 is a conserved active-site residue in the class A beta-lactamases, as well as other members of the serine penicillin-sensitive enzyme family; its role in catalysis remains controversial and uncertain. Mutation of Lys73 to alanine in the beta-lactamase from Bacillus licheniformis resulted in a substantial reduction in both turnover rate (k(cat)) and catalytic efficiency (k(cat)/K(m)), and a very significant shift in pK(1) to higher pH in the bell-shaped pH-rate profiles (k(cat)/K(m)) for several penicillin and cephalosporin substrates. The increase in pK(1) is consistent with the removal of the positive ammonium group of the lysine from the proximity of Glu166, to which the acid limb has been ascribed. The alkaline limb of the k(cat)/K(m) vs profiles is not shifted appreciably, as might have been expected if this limb reflected the ionization of Lys73 in the wild-type enzyme. The k(cat)/K(m) at the pH optimum for the mutant was down about 200-fold for penicillins and around 10(4) for cephalosporins, compared to the wild-type, suggesting significant differences in the mechanisms for catalysis of penicillins compared to cephalosporins. Burst kinetics were observed with several substrates assayed with K73A beta-lactamase, indicating an underlying branched-pathway kinetic scheme, and rate-limiting deacylation. FTIR analysis was used to determine whether acylation or deacylation was rate limiting. In general, acylation was the rate-limiting step for cephalosporin substrates, whereas deacylation was rate-limiting for penicillin substrates. The results indicate that Lys73 plays an important role in both the acylation and deacylation steps of the catalytic mechanism. The effects of this mutation (K73A) indicate that Lys73 does not function as a general base in the catalytic mechanism of beta-lactamase. The existence of bell-shaped pH-rate profiles for the K73A variant suggests that Lys73 is not directly responsible for either limb in such plots. It is likely that both Glu166 and Lys73 are important to each other in terms of maintaining the optimum electrostatic environment for fully efficient catalytic activity to occur. PMID- 10819960 TI - Comparative structural analysis and substrate specificity engineering of the hyperthermostable beta-glucosidase CelB from Pyrococcus furiosus. AB - The substrate specificity of the beta-glucosidase (CelB) from the hyperthermophilic archaeon Pyrococcus furiosus, a family 1 glycosyl hydrolase, has been studied at a molecular level. Following crystallization and X-ray diffraction of this enzyme, a 3.3 A resolution structural model has been obtained by molecular replacement. CelB shows a homo-tetramer configuration, with subunits having a typical (betaalpha)(8)-barrel fold. Its active site has been compared to the one of the previously determined 6-phospho-beta-glycosidase (LacG) from the mesophilic bacterium Lactococcus lactis. The overall design of the substrate binding pocket is very well conserved, with the exception of three residues that have been identified as a phosphate binding site in LacG. To verify the structural model and alter its substrate specificity, these three residues have been introduced at the corresponding positions in CelB (E417S, M424K, F426Y) in different combinations: single, double, and triple mutants. Characterization of the purified mutant CelB enzyme revealed that F426Y resulted in an increased affinity for galactosides, whereas M424K gave rise to a shifted pH optimum (from 5.0 to 6.0). Analysis of E417S revealed a 5-fold and a 3-fold increase of the efficiency of hydrolyzing o-nitrophenol-beta-D-galactopyranoside-6-phosphate, in the single and triple mutants, respectively. In contrast, their activity on nonphosphorylated sugars was largely reduced (30-300-fold). The residue at position E417 in CelB seems to be the determining factor for the difference in substrate specificity between the two types of family 1 glycosidases. PMID- 10819963 TI - Transmembrane movement of diether phospholipids in human erythrocytes and human fibroblasts. AB - We have synthesized spin-labeled (SL) and fluorescently labeled diacyl, 1-alkyl-2 acyl-, and di-alkyl glycerophospholipids. The sn-2 chain was a short chain with either a nitroxide group or a 7-nitro-2, 1,3-benzoxadiazol-4-yl (NBD). After incorporation in the exoplasmic leaflet of human erythrocytes, we found that SL phosphatidylcholine (PC) redistributed very slowly across the plasma membrane, less than 20% reaching the cytoplasmic leaflet in 3 h at 37 degrees C. In contrast, SL-phosphatidylserine (PS) accumulated on the cytoplasmic leaflet with the same plateau corresponding to 90% of the probes inside. The characteristic times for inward redistribution were different for the three PS analogues: at 37 degrees C, the t(1/2) for the diacyl, alkyl-acyl, and dialkyl compounds were 2.3, 3.5, and 41 min, respectively. ATP depletion or incubation with N-ethylmaleimide inhibited the rapid translocation of the PS derivatives. The diether PS bearing an NBD group translocated very slowly in human erythrocytes and no acceleration by ATP could be measured. On the other hand, in human fibroblasts, the diether NBD-PS and SL-PS were both transported from the exoplasmic to the cytoplasmic monolayer of the plasma membrane as it is the case for the transport of the respective diester PS analogues. These results prove that the ether bonds do not prevent completely PS binding and translocation by the aminophospholipid translocase despite a probable hindrance due to the ether linkage on the sn-2 chain. Because of the high stability of the ether linkage, SL and NBD diether analogues should be useful to investigate lipid traffic in cultured cells. PMID- 10819962 TI - 1H NMR structural characterization of a nonmitogenic, vasodilatory, ischemia protector and neuromodulatory acidic fibroblast growth factor. AB - A shortened genetically engineered form of acidic fibroblast growth factor (aFGF), that includes amino acids 28-154 of the full-length sequence (154 residues) plus Met in substitution of Leu27, does not induce cell division even though it is recognized by the cell membrane receptor, triggers the early mitogenic events, and retains the neuromodulatory, vasoactive, and cardio- and neuroprotective properties of the native full-length molecule. Taken together, these properties make this truncated aFGF a promising compound in the treatment of a wide assortment of neurological and cardiovascular pathologies where aFGF mitogenic activity is dispensable. Differences in biological activities between the shortened aFGF and the wild-type form have been attributed to lack of stability, and to the specific amino acid sequence missing at the N-terminus. Here we show that this shortened aFGF form has a three-dimensional structure even more stable than the wild-type protein at the mitogenic assay conditions; that this structure is similar to that of the wild type except at site 1 of interaction with the cell membrane receptor; that its lack of mitogenic activity cannot be attributed to the specific missing sequence; and that the vasodilatory activity of aFGF seems impaired by alterations of the three-dimensional structure of site 2 of interaction with the cell membrane receptor. PMID- 10819964 TI - Determination of the structural role of the linking moieties in the DNA binding of adozelesin. AB - Adozelesin (formerly U73975, The Upjohn Co.) is a monofunctional DNA alkylating analogue of the antitumor antibiotic (+)-CC-1065. Adozelesin consists of a cyclopropa[c]pyrrolo[3,2-e]indol-4(5H)-one (CPI) alkylating subunit of (+)-CC 1065 and a indole and benzofurans subunit replacing the more complex pyrroloindole B and C subunits, respectively, of (+)-CC-1065. Previous studies have shown that adozelesin forms a reversible covalent DNA duplex adduct via a reaction between the N3 of adenine and the cyclopropyl of the cyclopropapyrroloindole (CPI) subunit. Gel electrophoresis studies have shown that adozelesin, like all the monofunctional (CPI)-based antitumor antibiotics, has a sequence preference for 5'-TTA-3' [the asterisk () indicates covalently modified base]. Molecular-modeling studies have shown that the bound adozelesin ligand spans a total of five base pairs including the modified adenine. These studies have also indicated that, owing to the orientation of the ligand within the base minor groove, there should be an overall preference for sequences rich in A.T base pairs, thus avoiding steric crowding around the exocyclic NH(2) of any guanines present. In this study, we have prepared and studied, by high-field NMR and restrained molecular mechanics (rMM) and dynamics (rMD), the duplex adduct formed between adozelesin and 5'-CGTAAGCGCTTACG-3'. Previous molecular modeling studies suggested that this sequence should be less preferred, since the two GC base pairs should lead to extensive steric crowding within the adduct, and this hypothesis has, however, never been supported by DNA-footprinting data. (1)H NMR of the adozelesin duplex adduct has reveals that, although Watson-Crick base pairing is maintained throughout the DNA duplex, there is significant distortion around the central base pairs. This distortion is the result of strong hydrogen bonding between the amide linker of the indole and benzofuran subunits, and the carbonyl of a central thymine base and second, weaker, hydrogen bond to the exocyclic NH(2) of the central guanine was also observed. (1)H NMR and rMD also indicate that, to accommodate this hydrogen-bond system, the bound adozelesin is not positioned centrally within the minor groove but pushed toward the modified DNA strand. Previous studies on the dimeric CPI analogue bizelesin have indicated the important role the ureylene linker plays in the DNA binding. This study indicates that a similar situation exists in the reaction of adozelesin with double-stranded DNA and provides a possible explanation into the unpredicted sequence selectivity of these ligands. PMID- 10819966 TI - Side-chain interactions in the plastocyanin-cytochrome f complex. AB - Cytochrome f and plastocyanin are redox partners in the photosynthetic electron transfer chain. Electron transfer from cytochrome f to plastocyanin occurs in a specific short-lived complex. To obtain detailed information about the binding interface in this transient complex, the effects of binding on the backbone and side-chain protons of plastocyanin have been analyzed by mapping NMR chemical shift changes. Cytochrome f was added to plastocyanin up to 0.3 M equiv, and the plastocyanin proton chemical shifts were measured. Out of approximately 500 proton resonances, 86% could be observed with this method. Nineteen percent demonstrate significant chemical-shift changes and these protons are located in the hydrophobic patch (including the copper ligands) and the acidic patches of plastocyanin, demonstrating that both areas are part of the interface in the complex. This is consistent with the recently determined structure of the complex [Ubbink, M., Ejdeback, M., Karlsson, B. G., and Bendall, D. S. (1998) Structure 6, 323-335]. The largest chemical-shift changes are found around His87 in the hydrophobic patch, which indicates tight contacts and possibly water exclusion from this part of the protein interface. These results support the idea that electron transfer occurs via His87 to the copper in plastocyanin and suggest that the hydrophobic patch determines the specificity of the binding. The chemical shift changes in the acidic patches are significant but small, suggesting that the acidic groups are involved in electrostatic interactions but remain solvent exposed. The existence of small differences between the present data and those used for the structure may imply that the redox state of the metals in both proteins slightly affects the structure of the complex. The chemical-shift mapping is performed on unlabeled proteins, making it an efficient way to analyze effects of mutations on the structure of the complex. PMID- 10819965 TI - The X6 "thermostabilizing" domains of xylanases are carbohydrate-binding modules: structure and biochemistry of the Clostridium thermocellum X6b domain. AB - Many polysaccharide-degrading enzymes display a modular structure in which a catalytic module is attached to one or more noncatalytic modules. Several xylanases contain a module of previously unknown function (termed "X6" modules) that had been implicated in thermostability. We have investigated the properties of two such "thermostabilizing" modules, X6a and X6b from the Clostridium thermocellumxylanase Xyn10B. These modules, expressed either as discrete entities or as their natural fusions with the catalytic module, were assayed, and their capacity to bind various carbohydrates and potentiate hydrolytic activity was determined. The data showed that X6b, but not X6a, increased the activity of the enzyme against insoluble xylan and bound specifically to xylooligosaccharides and various xylans. In contrast, X6a exhibited no affinity for soluble or insoluble forms of xylan. Isothermal titration calorimetry revealed that the ligand-binding site of X6b accommodates approximately four xylose residues. The protein exhibited K(d) values in the low micromolar range for xylotetraose, xylopentaose, and xylohexaose; 24 microM for xylotriose; and 50 microM for xylobiose. Negative DeltaH and DeltaS values indicate that the interaction of X6b with xylooligosaccharides and xylan is driven by enthalpic forces. The three dimensional structure of X6b has been solved by X-ray crystallography to a resolution of 2.1 A. The protein is a beta-sandwich that presents a tryptophan and two tyrosine residues on the walls of a shallow cleft that is likely to be the xylan-binding site. In view of the structural and carbohydrate-binding properties of X6b, it is proposed that this and related modules be re-assigned as family 22 carbohydrate-binding modules. PMID- 10819967 TI - Active site residues of cis-2,3-dihydro-2,3-dihydroxybiphenyl dehydrogenase from Comamonas testosteroni strain B-356. AB - cis-2,3-dihydro-2,3-dihydroxybiphenyl dehydrogenase (BphB) from Comamonas testosteroni strain B-356 is the second enzyme of the biphenyl/polychlorinated biphenyl degradation pathway. Based on the crystal structure of a related BphB, three conserved residues, Ser142, Tyr155, and Lys159, have been suggested to function as a "catalytic triad" as for other members of the short-chain alcohol dehydrogenase/reductase (SDR) family. In this study, substitution of each triad residue was examined in BphB. At pH 9.0, turnover numbers relative to wild-type enzyme were as follows: Y155F, 0.1%; S142A, 1%; and K159A, 10%. Although the Michaelis constants of K159A and S142A for cis-2,3-dihydro-2,3-dihydroxybiphenyl increased about 20-fold, relatively little change was observed in the K(m) for dinucleotide. The K159A mutant, which showed little dehydrogenase activity at pH 7, was sharply activated by increasing the pH, reaching almost 25% of the activity of the wild-type enzyme at pH 9. 8. These three residues are therefore critical for BphB activity, as suggested by the crystal structure and similarity to other SDR family members. In addition, BphB showed a strong preference for NAD(+) over NADP(+), with a 260-fold higher specificity constant (k(cat)/K(m)). Evidence is presented that the inefficient use of NADP(+) by BphB might partly be due to the presence of an aspartate residue at position 36. PMID- 10819968 TI - Limited proteolysis as a structural probe of the soluble alpha-glycerophosphate oxidase from Streptococcus sp. AB - As reported previously [Parsonage, D., Luba, J., Mallett, T. C., and Claiborne, A. (1998) J. Biol. Chem. 273, 23812-23822], the flavoprotein alpha glycerophosphate oxidases (GlpOs) from a number of enterococcal and streptococcal sources contain a conserved 50-52 residue insert that is completely absent in the homologous alpha-glycerophosphate dehydrogenases. On limited proteolysis with trypsin, the GlpO from Streptococcus sp. (m = 67.6 kDa) is readily converted to two major fragments corresponding to masses of approximately 40 and 23 kDa. The combined application of sequence and mass spectrometric analyses demonstrates that the 40-kDa fragment represents the N-terminus of intact GlpO (Met1-Lys368; 40.5 kDa), while the 23-kDa band represents a C-terminal fragment (Ala405-Lys607; 22.9 kDa). Hence, limited proteolysis in effect excises most of the GlpO insert (Ser355-Lys404), indicating that this represents a flexible region on the protein surface. The active-site and other spectroscopic properties of the enzyme, including both flavin and tryptophan fluorescence spectra, titration behavior with both dithionite and sulfite, and preferential binding of the anionic form of the oxidized flavin, were largely unaffected by proteolysis. Enzyme-monitored turnover analyses of the intact and nicked streptococcal GlpOs (at [GlpO] approximately 10 microM) demonstrate that the single major catalytic defect in the nicked enzyme corresponds to a 20-fold increase in K(m)(Glp); the basis for this altered kinetic behavior is derived from an 8-fold decrease in the second order rate constant for reduction of the nicked enzyme, as measured in anaerobic stopped-flow experiments. These results indicate that the flexible surface region represented by elements of the GlpO insert plays an important role in mediating efficient flavin reduction. PMID- 10819969 TI - The onset of the deuterium isotope effect in cytochrome c oxidase. AB - We have investigated the dynamics of proton equilibration within the proton transfer pathway of cytochrome c oxidase from bovine heart that is used for the transfer of both substrate and pumped protons during reaction of the reduced enzyme with oxygen (D-pathway). The kinetics of the slowest phase in the oxidation of the enzyme (the oxo-ferryl --> oxidized transition, F --> O), which is associated with proton uptake, were studied by monitoring absorbance changes at 445 nm. The rate constant of this transition, which is 800 s(-)(1) in H(2)O (at pH approximately 7.5), displayed a kinetic deuterium isotope effect of approximately 4 (i.e., the rate was approximately 200 s(-)(1) in 100% D(2)O). To investigate the kinetics of the onset of the deuterium isotope effect, fully reduced, solubilized CO-bound cytochrome c oxidase in H(2)O was mixed rapidly at a ratio of 1:5 with a D(2)O buffer saturated with oxygen. After a well-defined time period, CO was flashed off using a short laser flash. The time between mixing and flashing off CO was varied within the range 0. 04-10 s. The results show that for the bovine enzyme, the onset of the deuterium isotope effect takes place within two time windows of O transition is internal proton transfer from a site, proposed to be Glu (I-286) (R. sphaeroides amino acid residue numbering), to the binuclear center. The spontaneous equilibration of protons/deuterons with this site in the interior of the protein is slow (approximately 1 s). PMID- 10819970 TI - Synchronization of the three reaction centers within carbamoyl phosphate synthetase. AB - Carbamoyl phosphate synthetase from E. coli catalyzes the synthesis of carbamoyl phosphate through a series of four reactions occurring at three active sites connected by a molecular tunnel of 100 A. To understand the mechanism for coordination and synchronization among the active sites, the pre-steady-state time courses for the formation of phosphate, ADP, glutamate, and carbamoyl phosphate were determined. When bicarbonate and ATP were rapidly mixed with CPS, a stoichiometric burst of acid-labile phosphate and ADP was observed with a formation rate constant of 1100 min(-)(1). The burst phase was followed by a linear steady-state phase with a rate constant of 12 min(-)(1). When glutamine or ammonia was added to the initial reaction mixture, the magnitude and the rate of formation of the burst phase for either phosphate or ADP were unchanged, but the rate constant for the linear steady-state phase increased to an average value of 78 min(-)(1). These results demonstrate that the initial phosphorylation of bicarbonate is independent of the binding or hydrolysis of glutamine. The pre steady-state time course for the hydrolysis of glutamine in the absence of ATP exhibited a burst of glutamate formation with a rate constant of 4 min(-)(1) when the reaction was quenched with base. In the presence of ATP and bicarbonate, the rate constant for the formation of the burst of glutamate was 1100 min(-)(1). The hydrolysis of ATP thus enhanced the hydrolysis of glutamine by a factor of 275, but there was no effect by glutamine on the initial phosphorylation of bicarbonate. The pre-steady-state time course for the formation of carbamoyl phosphate was linear with an overall rate constant of 72 min(-)(1). The absence of an initial burst of carbamoyl phosphate formation eliminates product release as a rate-determining step for CPS. Overall, these results have been interpreted to be consistent with a mechanism whereby the phosphorylation of bicarbonate serves as the initial trigger for the rest of the reaction cascade. The formation of the carboxy phosphate intermediate within the large subunit must induce a conformational change to the active site of the small subunit that enhances the hydrolysis of glutamine. Thus, ammonia is not released into the molecular tunnel until the activated bicarbonate is ready to form carbamate. The rate-limiting step for the steady-state assembly of carbamoyl phosphate is either the formation, migration, or phosphorylation of the carbamate intermediate. PMID- 10819971 TI - Asymmetry in the autophosphorylation of the two-component regulatory system transmitter protein nitrogen regulator II of Escherichia coli. AB - Autophosphorylation of the homodimeric two-component system transmitter protein nitrogen regulator II (NRII; also NtrB) of Escherichia coli is the first step in the activation of nitrogen-regulated (Ntr) gene transcription. We show that the autophosphorylation of NRII was asymmetric, with phosphorylation of the first and second subunits of the dimer displaying different equilibria (under our experimental conditions K(1) approximately 0. 345, K(2) approximately 0.0044). Phosphorylation of both subunits of NRII was rapid, but the very rapid reversal of the phosphorylation of the second subunit was responsible for the equilibrium position of the reaction. Complete phosphorylation of NRII was only observed under conditions where ADP, a product of the autophosphorylation reaction, was removed by an enzymatic system. Purified, doubly phosphorylated NRII (NRII approximately P(2)) was stable in the absence of nucleotides at 0 degrees C but was dephosphorylated to the hemiphosphorylated form at 37 degrees C. In the presence of a low concentration of ADP, half of the phosphoryl groups from NRII approximately P(2) were rapidly dephosphorylated, while the remaining phosphoryl groups were slowly dephosphorylated. Experiments with heterodimers containing wild-type and mutant, nonphosphorylatable subunits suggested that the asymmetry of NRII autophosphorylation was not preexisting but resulted from the autophosphorylation of one subunit. PMID- 10819972 TI - Interaction of NADP(H) with oxidized and reduced P450 reductase during catalysis. Studies with nucleotide analogues. AB - Previous studies have shown that the interaction of P450 reductase with bound NADP(H) is essential to ensure fast electron transfer through the two flavin cofactors. In this study we investigated in detail the interaction of the house fly flavoprotein with NADP(H) and a number of nucleotide analogues. 1,4,5,6 Tetrahydro-NADP, an analogue of NADPH, was used to characterize the interaction of P450 reductase with the reduced nucleotide. This analogue is inactive as electron donor, but its binding affinity and rate constant of release are very close to those for NADPH. The 2'-phosphate contributes about 5 kcal/mol of the binding energy of NADP(H). Oxidized nicotinamide does not interact with the oxidized flavoprotein, while reduced nicotinamide contributes 1.3 kcal/mol of the binding energy. Oxidized P450 reductase binds NADPH with a K(d) of 0.3 microM, while the affinity of the reduced enzyme is considerably lower, K(d) = 1.9 microM. P450 reductase catalyzes a transhydrogenase reaction between NADPH and oxidized nucleotides, such as thionicotinamide-NADP(+), acetylpyridine-NADP(+), or [(3)H]NADP(+). The reverse reaction, reduction of [(3)H]NADP(+) by the reduced analogues, is also catalyzed by P450 reductase. We define the mechanism of the transhydrogenase reaction as follows: NADPH binding, hydride ion transfer, and release of the NADP(+) formed. An NADP(+) or its analogue binds to the two electron-reduced flavoprotein, and the electron-transfer steps reverse to transfer hydride ion to the oxidized nucleotide, which is released. Measurements of the flavin semiquinone content, rate constant for NADPH release, and transhydrogenase turnover rates allowed us to estimate the steady-state distribution of P450 reductase species during catalysis, and to calculate equilibrium constants for the interconversion of catalytic intermediates. Our results demonstrate that equilibrium redox potentials of the flavin cofactors are not the sole factor governing rapid electron transfer during catalysis, but conformational changes must be considered to understand P450 reductase catalysis. PMID- 10819973 TI - Suppression of microtubule dynamic instability and treadmilling by deuterium oxide. AB - Deuterium oxide (D(2)O) is known to promote the assembly of tubulin into microtubules in vitro, to increase the volume of mitotic spindles and the number and length of spindle microtubules, and to inhibit mitosis. Reasoning that its actions on cellular microtubules could be due to modulation of microtubule dynamics, we examined the effects of replacing H(2)O with D(2)O on microtubule dynamic instability, treadmilling, and steady-state GTPase activity. We found that replacing 50% or more of the H(2)O with D(2)O promoted microtubule polymerization and stabilized microtubules against dilution-induced disassembly. Using steady-state axoneme-seeded microtubules composed of pure tubulin and video microscopy, we found that 84% D(2)O decreased the catastrophe frequency by 89%, the shortening rate by 80%, the growing rate by 50%, and the dynamicity by 93%. Sixty percent D(2)O decreased the treadmilling rate of microtubules composed of tubulin and microtubule-associated proteins by 42%, and 89% D(2)O decreased the steady-state GTP hydrolysis rate by 90%. The mechanism responsible for the ability of D(2)O to stabilize microtubule dynamics may involve enhancement of hydrophobic interactions in the microtubule lattice and/or the substitution of deuterium bonds for hydrogen bonds. PMID- 10819974 TI - Partial IGF affinity of circulating N- and C-terminal fragments of human insulin like growth factor binding protein-4 (IGFBP-4) and the disulfide bonding pattern of the C-terminal IGFBP-4 domain. AB - Within the IGF axis, the insulin-like growth factor-binding proteins (IGFBPs) are known to play a pivotal role in cell proliferation and differentiation. Defined proteolysis of the IGFBPs is proposed to be an essential mechanism for regulating IGF bioavailability. The generated IGFBP fragments in part exhibit different IGF dependent and -independent biological activities. Characterizing naturally occurring forms of IGFBPs in human plasma, we identified both a N- and a C terminal fragment of IGFBP-4 by means of immunoreactivity screening. As a source for peptide isolation, we used large amounts of human hemofiltrate obtained from patients with chronic renal failure. Purification of the IGFBP-4 peptides from hemofiltrate was performed by consecutive cation-exchange and reverse-phase chromatographic steps. Mass spectrometric and sequence analysis revealed an M(r) of 13 233 for the purified N-terminal fragment spanning residues Asp(1)-Phe(122) of IGFBP-4 and an M(r) of 11 344 for the C-terminal fragment extending from Lys(136) to Glu(237). Proteolytic digestion and subsequent biochemical analysis showed that the six cysteines of the C-terminal IGFBP-4 fragment are linked between residues 153-183, 194-205, and 207-228 (disulfide bonding pattern, 1-2, 3 4, and 5-6). Plasmon resonance spectroscopy, ligand blot analysis, and saturation and displacement studies demonstrated a very low affinity of the C-terminal IGFBP 4 fragment for the IGFs (IGF-II, K(d) = 690 nM; IGF-I, K(d) > 60 nM), whereas the N-terminal fragment retained significant IGF binding properties (IGF-II, K(d) = 17 nM; IGF-I, K(d) = 5 nM). This study provides the first molecular characterization of circulating human IGFBP-4 fragments formed in vivo exhibiting an at least 5-fold decrease in the affinity of the N-terminal IGFBP-4 fragment for the IGFs and a very low IGF binding capacity of the C-terminal fragment. PMID- 10819975 TI - Binding of IRS proteins to calmodulin is enhanced in insulin resistance. AB - The IRS proteins, major endogenous targets of the insulin receptor, bind to calmodulin in a Ca(2+)-dependent manner. Here, we have examined the interaction between these proteins in animal and cultured cell models of insulin resistance. Both IRS-1 and IRS-2 co-immunoprecipitate with calmodulin from insulin target tissues in rats. The interaction between calmodulin and IRS proteins in rat soleus muscle was enhanced when insulin resistance was induced in rats by treatment with dexamethasone for 5 days. Moreover, injection of angiotensin II into the inferior vena cava enhanced the binding in rat cardiac muscle. Similarly, increased binding between calmodulin and IRS-1 was observed in isolated cells incubated with tumor necrosis factor-alpha. Overexpression of calmodulin in Chinese hamster ovary cells reduced the tyrosine phosphorylation of IRS-1 induced by insulin, with a concomitant decrease in insulin-stimulated association of IRS-1 with the 85-kDa regulatory subunit of phosphatidylinositol 3 kinase. Insulin-stimulated phosphatidylinositol 3-kinase activity associated with IRS-1 was also reduced in cells overexpressing calmodulin, while this activity was increased in cells incubated with the cell-permeable calmodulin antagonist trifluoperazine. These data demonstrate an enhanced interaction between calmodulin and IRS proteins in models of insulin resistance and suggest a possible mechanism by which increased intracellular Ca(2+) concentrations may contribute to impaired insulin sensitivity. PMID- 10819976 TI - Ligand recognition by the lactose permease of Escherichia coli: specificity and affinity are defined by distinct structural elements of galactopyranosides. AB - Specificity of substrate recognition in lactose permease is directed toward the galactosyl moiety of lactose. In this study, binding of 31 structural analogues of D-galactose was examined by site-directed N-[(14)C]ethylmaleimide-labeling of the substrate-protectable Cys148 in the binding site. Alkylation of Cys148 is blocked by D-galactose with an apparent affinity of approximately 30 mM. Epimers of D-galactose at C-3 (D-gulose) and C-4 (D-glucose) or deoxy derivatives at these positions exhibit no binding whatsoever, indicating that these OH groups participate in essential interactions. Interestingly, the C-2 epimer alpha-D talose binds almost as well as D-galactose, while 2-deoxy-D-galactose affords no substrate protection, indicating that nonstereospecific H-bonding at C-2 is required for stable binding. No substrate protection is detected with D-fucose, L arabinose, 6-deoxy-6-fluoro-D-galactose, 6-O-methyl-D-galactose, or D galacturonic acid, suggesting that the C-6 OH is an essential H-bond donor. Both alpha- and beta-methyl D-galactopyranosides bind more strongly than galactose, supporting the notion that the cyclic pyranose conformation is the bound form and that the anomeric configuration at C-1 does not contribute to substrate specificity. However, methyl or allyl alpha-D-galactopyranosides exhibit 60-fold lower apparent K(d)'s than D-galactose, demonstrating that binding affinity is significantly influenced by the functional group at C-1 and its orientation. Taken together, the observations confirm and extend the current binding site model [Venkatesan, P., and Kaback, H. R. (1998) Proc. Natl. Acad. Sci. U.S.A. 95, 9802-9807] and indicate that specificity toward galactopyranosides is governed by H-bonding interactions at C-2, C-3, C-4, and C-6 OH groups, while binding affinity can be increased dramatically by hydrophobic interactions with the nongalactosyl moiety. PMID- 10819977 TI - Kinetic analysis of the role of zinc in the interaction of domain 5 of high molecular weight kininogen (HK) with heparin. AB - Previous investigations have shown that HK and its light chain bind heparin, preventing the enhancement of antithrombin inhibition of thrombin and potentiating the inhibition of plasma kallikrein by antithrombin. We found that both HK and HKa bound heparin, but HK exhibited a greater affinity. We therefore localized the binding sites for heparin on HK. HK domains 5 and 6 of the light chain as well as domain 3 from the heavy chain, expressed as glutathione S transferase (GST) fusion proteins in Escherichia coli, were tested for binding to immobilized heparin by surface plasmon resonance using a BiaCore 2000 instrument. GST-D5, but not GST-D3, GST-D6, or GST, bound to heparin when the recombinant domains were present at a concentration of 70 nM. To localize more precisely the amino acid sequences on D5, both of the subdomains, histidine-glycine-rich GST (K420-D474) and histidine-glycine-lysine-rich GST-(H475-S626), were expressed and tested for binding to immobilized heparin. The K(d) was much lower for GST-(K420 D474) than for GST-(H475-S626) in the presence or absence of Zn(2+). GST-(K420 D474) was effective in decreasing the rate of inactivation of thrombin by antithrombin in the presence of heparin and Zn(2+), while GST-(H475-S626) had no effect. We conclude that the binding of heparin to HK is a complex function of Zn(2+) interacting with histidines in the sequence K420-D474 to create high affinity binding sites. HK has the potential to be an important modulator of heparin therapy. PMID- 10819978 TI - Heterotrimeric collagen peptides as fluorogenic collagenase substrates: synthesis, conformational properties, and enzymatic digestion. AB - The collagenase cleavage site of collagen type I, i.e., the sequence portions 772 784 (P(4)-P(9)') and 772-785 (P(4)-P(10)') of the two alpha1-chains and the sequence portion 772-784 (P(4)-P(9)') of the alpha2-chain, were assembled in an alpha1alpha2alpha1' register by C-terminal cross-linking of these peptides with an artificial cystine knot. The triple-helical conformation of the construct was stabilized by N-terminal extensions with (Gly-Pro-Hyp)(5) repeats. The gaps in the sequence alignment were filled up, and the alpha1-chain was dansylated and the alpha1'-chain was acylated with a tryptophan residue to place in spatial proximity the two chromophores for an efficient fluorescence resonance energy transfer. Although the incorporation of the two N-terminal chromophores leads to partial destabilization of the overall triple-helical fold, the heterotrimer behaved as a collagen-like substrate of the matrix metalloproteinases MMP-1 and MMP-13. Cleavage of the fluorogenic heterotrimer leads to a 6-fold increase in fluorescence intensity, thus making it a useful fluorogenic substrate for interstitial collagenases. With this folded heterotrimeric collagen molecule it was shown that fluorescence resonance energy transfer, as applied so far only for the design of linear fluorogenic enzyme substrates, can also be exploited in conformation dependency. PMID- 10819979 TI - A hemerythrin-like domain in a bacterial chemotaxis protein. AB - Hemerythrin (Hr) is an O(2)-carrying protein found in some marine invertebrates. A conserved sequence motif in all Hrs provides five histidine and two carboxylate ligands to an oxo-/hydroxo-bridged diiron active site, as well as a hydrophobic O(2) binding pocket. Database searches located a previously unrecognized Hr-like sequence motif at the 3' end of the gene, dcrH, from the anaerobic sulfate reducing bacterium, Desulfovibrio (D.) vulgaris (Hildenborough). This gene encodes a putative methyl-accepting chemotaxis protein, DcrH. We have established by immunoblotting that a full-length DcrH, including the Hr-like domain, is expressed in D. vulgaris (Hildenborough). The C-terminal domain of DcrH, when expressed separately in recombinant form in Escherichia coli, was found to fold into a stable protein, DcrH-Hr. The UV-vis absorption and resonance Raman spectra of DcrH-Hr, and of its azide adduct, provide clear evidence for an oxo-bridged diiron(III) site very similar to that found in Hr. Based on UV-vis absorption spectra, exposure of the reduced (colorless, presumably diferrous) DcrH-Hr to air resulted in formation of an O(2) adduct also very similar to that of Hr. Unlike that of Hr, the O(2) adduct of DcrH-Hr autoxidized within a few minutes at room temperature. The O(2) binding pocket of DcrH-Hr appears to be larger than that of Hr. Given the air-sensitive nature of D. vulgaris and the putative chemotactic function of DcrH, one possible role for the Hr-like domain of DcrH is O(2) sensing. DcrH-Hr is the first characterized example of a Hr-like protein from any microorganism. PMID- 10819980 TI - Inhibition of gene expression and cell proliferation by triple helix-forming oligonucleotides directed to the c-myc gene. AB - Triple helix-forming oligonucleotides (TFOs) bind with high affinity and specificity to homopurine-homopyrimidine sequences in DNA and have been shown to inhibit transcription of target genes in various experimental systems. In the present study, we evaluated the ability of 3'-amino-modified phosphodiester TFOs directed to four sites in the c-myc gene to inhibit gene expression and proliferation of human leukemia (CEM, KG-1, and HL-60) and lymphoma (Raji and ST486) cells. GT-rich TFOs were designed to target sequences located either upstream (myc1 and -2) or downstream (myc3 and -4) of the P2 promoter, which is the major c-myc promoter. Myc2, which was directed to a site immediately upstream of this promoter, inhibited c-myc expression and proliferation of CEM cells. The effects of this TFO were sequence- and target-specific, since control oligonucleotides and TFOs directed to other sites were less or not active. Myc2 was also effective in KG-1, HL-60, and Raji cells. In contrast, ST486 cells were more sensitive to myc3, which targets a sequence in intron 1 upstream of the P3 promoter, than myc2. As result of a chromosomal translocation, P3 is the active promoter in ST486 cells. This study demonstrates the activity and specificity of TFOs designed to act as repressors of c-myc gene expression in human leukemia and lymphoma cells. Our results suggest that this is a valid approach to selectively inhibit gene expression and cancer cell growth, and encourage further investigation of its potential applications in cancer therapy. PMID- 10819981 TI - Differential effects of N-acetyl-2-aminofluorene and N-2-aminofluorene adducts on the conformational change in the structure of DNA polymerase I (Klenow fragment). AB - The carcinogen N-acetyl-2-aminofluorene forms two major DNA adducts: the N-(2' deoxyguanosin-8-yl)-2-acetylaminofluorene adduct (dG-C8-AAF) and its deacetylated derivative, the N-(2'-deoxyguanosin-8-yl)-2-aminofluorene adduct (dG-C8-AF). It is well established that the AAF adduct is a very strong block for DNA synthesis in vitro while the AF adduct is more easily bypassed. In an effort to understand the molecular mechanism of this phenomenon, the structure of the complex of an exonuclease-deficient Escherichia coli DNA polymerase I (Klenow fragment) bound to primer-templates containing either an AF or AAF adduct in or near the active site was probed by nuclease and protease digestion analyses. The results of these experiments suggest that positioning the AAF adduct in the polymerase active site strongly inhibits the conformational change that is required for the insertion of a nucleotide. Similar experiments with AF-modified primer-templates shows a much less pronounced effect. The inhibition of the conformational change by either adduct is not detected if they are positioned in the single-stranded part of the template just one nucleotide before the active site. These findings may explain the different abilities of these lesions to block DNA synthesis. PMID- 10819982 TI - Statistical conformation of human plasma fibronectin. AB - Fibronectin is a multifunctional glycoprotein (molecular mass, M = 530 kg/mol) of the extra cellular matrix (ECM) having a major role in cell adhesion. In physiological conditions, the conformation of this protein still remains debated and controversial. Here, we present a set of results obtained by scattering experiments. In "native" conditions, the radius of gyration (R(g) = 15.3 +/- 0.3 nm) was determined by static light scattering as well as small-angle neutron scattering. The hydrodynamic radius (R(H) = 11.5 +/- 0.1 nm) was deduced from quasi-elastic light scattering measurements. These results imply a low internal concentration compared to that of usual globular proteins. This is also confirmed by the ratio R(H)/R(g) = 0. 75 +/- 0.02 consistent with a Gaussian chain, whereas R(H)/R(g) = 1. 3 for spherical shaped molecules. However, adding a denaturing agent (urea 8 M) increases R(g) by a factor 2. This means that fibronectin "native" chain is not either completely unfolded. The average shape of fibronectin conformation was also probed by small-angle neutron scattering performed for reverse scattering vector q(-)(1) smaller than R(g) (0.2 < q(-)(1) < 15 nm). The measured form factor is in complete agreement with the form factor of a random string of 56 beads of 5 nm diameter. It rules out the possibility of unfolded chain as well as globular structures. These results have structural and biological implications as far as ECM organization is concerned. PMID- 10819983 TI - Valine of the YVDD motif of moloney murine leukemia virus reverse transcriptase: role in the fidelity of DNA synthesis. AB - The YXDD motif is highly conserved in the reverse transcriptase family. The variable X residue is occupied by valine and methionine in MuLV RT and HIV-1 RT, respectively. Previous studies have shown that Tyr 222, the Y residue of the YXDD motif in MuLV RT, constitutes a major component of the fidelity center of the enzyme [Kaushik, N., Singh, K., Alluru, I., and Modak, M. J. (1999) Biochemistry 38, 2617-2627]. In this work, we present evidence that reverse transcriptases containing valine in the "X" position of the YXDD motif generally catalyze DNA synthesis with greater fidelity than those containing methionine or alanine. In the MuLV RT system, the two mutants V223M and V223A exhibited an overall reduced fidelity of DNA synthesis, specifically for RNA-templated reactions. Further analysis revealed that these mutants exhibit a higher efficiency of misinsertion on MS2 RNA than the wild-type enzyme for every mispair tested. However, unlike HIV-1 RT, the insensitivity of the wild-type MuLV RT to all four ddNTPs remained unchanged by mutation of V223 to Met or Ala. A 3D molecular model of the ternary complex of MuLV RT, template primer, and dNTP suggests that Val 223 along with its neighboring Tyr 222 stabilizes the substrate binding pocket via hydrophobic interactions with the dNTP substrate and template-primer. PMID- 10819984 TI - Strand-separating activity of hepatitis C virus helicase in the absence of ATP. AB - HCV helicase [E(wt)] catalyzed strand separation of a short DNA duplex (F21:HF31) formed from a 5'-hexachlorofluorescein-tagged 31-mer (HF31) and a 3'-fluorescein tagged 21-mer (F21) complementary to the 5'-end of HF31. Strand separation was monitored by the fluorescence increase associated with the formation of F21 from F21:HF31. In the presence of ATP, the strand-separating activity was catalytic. In the absence of ATP and with E(wt) concentrations greater than that of F21:HF31, a biphasic fluorescence increase was observed at 25 degrees C. The late phase of this reaction was assigned to the separation of F21 from F21:HF31. The ATP-independent strand-separating reaction occurred more rapidly in the absence of Mg(2+) than in its presence. This result correlated with a lower T(m) value of F21:HF31 in the absence of 3.5 mM Mg(2+) than in its presence (45 vs 63 degrees C). The stoichiometry for the strand-separating reaction in the absence of ATP was 8 mol of E(wt) per mole of F21:HF31 separated into single-stranded F21 and HF31. The dissociation constants of HCV helicase for F21, HF31, and F21:HF31 in the absence of Mg(2+) were 0.6 +/- 0.4, 6 +/- 1, and 7.3 +/- 0.9 nM, respectively. Histidinyl-tagged E(wt) [hE(wt)] and a mutant enzyme [hE(V432A)] were prepared. hE(wt) and E(wt) bound F21 and HF31 with similar affinities and had similar ATP-dependent helicase activities, whereas hE(V432A) bound F21 and HF31 with affinities similar to that of E(wt) but had greatly reduced ATP dependent helicase activities. In contrast to E(wt) and hE(wt), hE(V432A) did not support the ATP-independent strand-separating reaction. Consequently, the ATP independent strand-separating reaction was not only the result of the high affinity of the enzyme for single-stranded DNA. The enzyme preferentially used duplex DNA with a 3'-tail for the ATP-dependent helicase reaction. In contrast, the enzyme strand-separated blunt-ended, 5'-tailed, and 3'-tailed duplex DNA equally effectively in the ATP-independent strand-separating reaction. PMID- 10819985 TI - Kinetic analysis of the effects of mutagenesis of W501 and V432 of the hepatitis C virus NS3 helicase domain on ATPase and strand-separating activity. AB - Two hydrophobic residues, W501 and V432, in the nucleic acid (NA) binding pocket of the HCV helicase domain (E) were mutagenized in an effort to investigate contributions of these residues to substrate affinities and to enzymatic activities. The affinities of wild-type [hE(wt)] and mutant enzymes [hE(W501F), hE(W501A), and hE(V432A)] for NA and ATP were determined by monitoring changes in the intrinsic protein fluorescence, in the fluorescence of fluorescently tagged nucleic acid, and in the enzymatic activity. The steady-state kinetic parameters of the mutant enzymes for ATP hydrolysis (at saturating concentrations of NA) were similar to those of hE(wt). hE(W501F), hE(W501A), and hE(V432A) had strand separating activities that were 136%, 3.8%, and 3.1% of that of hE(wt). The processivities of hE(W501F), hE(W501A), and hE(V432A) were reduced relative to that of hE(wt). The reduced processivities of hE(W501F) and hE(W501A) were primarily due to an increase in the rate of dissociation of E. ATP from E.ATP.NA. The reduced processivity of hE(V432A) was primarily due to a reduction in the intrinsic forward rate constant for strand separation. This result suggested that V432 may constitute part of the forward "stepping" motor of E. hE(W501A) and hE(V432A) did not display a dominant negative phenotype in a steady-state helicase assay with hE(wt). hE(wt) stored in the presence of beta-mercaptoethanol was covalently modified at three cysteinyl residues. The biological significance of the potential reactivity of these cysteinyl residues on hE(wt) is unknown. PMID- 10819986 TI - Peridinin chlorophyll a protein: relating structure and steady-state spectroscopy. AB - Peridinin chlorophyll a protein (PCP) from Amphidinium carterae has been studied using absorbance (OD), linear dichroism (LD), circular dichroism (CD), fluorescence emission, fluorescence anisotropy, fluorescence line narrowing (FLN), and triplet-minus-singlet spectroscopy (T-S) at different temperatures (4 293 K). Monomeric PCP binds eight peridinins and two Chls a. The trimeric structure of PCP, resolved at 2 A [Hofmann et al. (1996) Science 27, 1788-1791], allows modeling of the Chl a-protein and Chl a-Chl a interactions. The FLN spectrum shows that Chl a is not or is very weakly hydrogen-bonded and that the central magnesium of the emitting Chl a is monoligated. Simulation of the temperature dependence of the absorption spectra indicates that the Huang-Rhys factor, characterizing the electron-phonon coupling strength, has a value of approximately 1. The width of the inhomogeneous distribution function is estimated to be 160 cm(-)(1). LD experiments show that the two Chls a in PCP are essentially isoenergetic at room temperature and that a substantial amount of PCP is in a trimeric form. From a comparison of the measured and simulated CD, it is concluded that the interaction energy between the two Chls a within one monomer is very weak, <10 cm(-)(1). In contrast, the Chls a appear to be strongly coupled to the peridinins. The 65 cm(-)(1) band that is visible in the low-frequency region of the FLN spectrum might indicate a Chl a-peridinin vibrational mode. The efficiency of Chl a to peridinin triplet excitation energy transfer is approximately 100%. On the basis of T-S, CD, LD, and OD spectra, a tentative assignment of the peridinin absorption bands has been made. PMID- 10819987 TI - Biochemical characterization of rat P450 2C11 fused to rat or bacterial NADPH P450 reductase domains. AB - cDNAs coding for rat P450 2C11 fused to either a bacterial (the NADPH-cytochrome P450 BM3 reductase domain of P450 BM3) or a truncated form of rat NADPH-P450 reductases were expressed in Escherichia coli and characterized enzymatically. Measurements of NADPH cytochrome c reductase activity showed fusion-dependent increases in the rates of cytochrome c reduction by the bacterial or the mammalian flavoprotein (21 and 48%, respectively, of the rates observed with nonfused enzymes). Neither the bacterial flavoprotein nor the truncated rat reductase supported arachidonic acid metabolism by P450 2C11. In contrast, fusion of P450 2C11 to either reductase yielded proteins that metabolized arachidonic acid to products similar to those obtained with reconstituted systems containing P450 2C11 and native rat P450 reductase. Addition of a 10-fold molar excess of rat P450 reductase markedly increased the rates of metabolism by both fused and nonfused P450s 2C11. These increases occurred with preservation of the regioselectivity of arachidonic acid metabolism. The fusion-independent reduction of P450 2C11 by bacterial P450 BM3 reductase was shown by measurements of NADPH dependent H(2)O(2) formation [73 +/- 10 and 10 +/- 1 nmol of H(2)O(2) formed min( )(1) (nmol of P450)(-)(1) for the reconstituted and fused protein systems, respectively]. These studies demonstrate that (a) a self-sufficient, catalytically active arachidonate epoxygenase can be constructed by fusing P450 2C11 to mammalian or bacterial P450 reductases and (b) the P450 BM3 reductase interacts efficiently with mammalian P450 2C11 and catalyzes the reduction of the heme iron. However, fusion is required for metabolism and product formation. PMID- 10819988 TI - Iron-sulfur cluster interconversions in biotin synthase: dissociation and reassociation of iron during conversion of [2Fe-2S] to [4Fe-4S] clusters. AB - Biotin synthase catalyzes the insertion of a sulfur atom into the saturated C6 and C9 carbons of dethiobiotin. This reaction has long been presumed to occur through radical chemistry, and recent experimental results suggest that biotin synthase belongs to a family of enzymes that contain an iron-sulfur cluster and reductively cleave S-adenosylmethionine, forming an enzyme or substrate radical, 5'-deoxyadenosine, and methionine. Biotin synthase (BioB) is aerobically purified as a dimer of 38 kDa monomers that contains two [2Fe-2S](2+) clusters per dimer. Maximal in vitro biotin synthesis requires incubation of BioB with dethiobiotin, AdoMet, reductants, exogenous iron, and crude bacterial protein extracts. It has previously been shown that reduction of BioB with dithionite in 60% ethylene glycol produces one [4Fe-4S](2+/1+) cluster per dimer. In the present work, we use UV/visible and electron paramagnetic resonance spectroscopy to show that [2Fe 2S] to [4Fe-4S] cluster conversion occurs through rapid dissociation of iron from the protein followed by rate-limiting reassociation. While in 60% ethylene glycol the product of dithionite reduction is one [4Fe-4S](2+) cluster per dimer, the product in water is one [4Fe-4S](1+) cluster per dimer. Further, incubation with excess iron, sulfide, and dithiothreitol produces protein that contains two [4Fe 4S](2+) clusters per dimer; subsequent reduction with dithionite produces two [4Fe-4S](1+) clusters per BioB dimer. BioB that contains two [4Fe-4S](2+/1+) clusters per dimer is rapidly and reversibly reduced and oxidized, suggesting that this is the redox-active form of the iron-sulfur cluster in the anaerobic enzyme. PMID- 10819989 TI - Lecithin retinol acyltransferase contains cysteine residues essential for catalysis. AB - Lecithin retinol acyltransferase (LRAT) is an essential enzyme in vitamin A metabolism and mobilization. The membrane-bound enzyme catalyzes the transfer of an acyl group from the sn-1 position of lecithin to vitamin A to generate retinyl esters. The sequence of LRAT is novel and hence does not suggest a mechanistic class to which the enzyme belongs. However, the activity of the enzyme is exceedingly sensitive to affinity labeling and group-specific reagents directed toward thiol groups. LRAT from human retinal pigment epithelium has cysteine residues at positions 161, 168, 182, and 208. Site-specific mutagenic studies show that C182 and C208 can be converted to alanines with little affect on activity. The activities of the C161A and C168A mutants are virtually nil. Moreover, while C168S is substantially active, C161S possesses only a few percent of the activity of wild-type (WT) LRAT. Also, pH-rate profiles show that C168S has virtually the same profile as WT LRAT, while C161S shows an aberrant profile quite unlike that of WT LRAT. Therefore, LRAT is a thiol acyltransferase and C161 may be the essential nucleophilic residue critical for catalysis. PMID- 10819990 TI - Cyclic peptides incorporating 4-carboxyphenylalanine and phosphotyrosine are potent inhibitors of pp60(c-)(src). AB - The protein tyrosine kinase, pp60(c-)(src), is involved in cellular signaling and is activated during mitosis and in various tumors. We have been employing cyclic decapeptides to identify the determinants for substrate binding and phosphorylation to develop inhibitors competitive with protein substrates of Src. A structure-activity study [McMurray, J. S., Budde, R. J. A., Ke, S., Obeyesekere, O. U., Wang, W., Ramdas, L., and Lewis, C. A. (1998) Arch. Biochem. Biophys. 355, 124] revealed that, at the position 3 residues C-terminal to the phosphorylated tyrosine (Y + 3), both glutamic acid and phenylalanine gave identical K(i), K(m), and V(max) values. We hypothesized that the area of Src that binds the Y + 3 residue contains either a positively charged lysine or an arginine, capable of ionic interactions with glutamic acid or cation-pi interactions with phenylalanine. To test this hypothesis, a series of phenylalanine analogues were substituted at position 7 (the Y + 3 residue) in cyclo(Asp(1)-Asn(2)-Glu(3)-Tyr(4)-Ala(5)-Phe(6)-Phe(7)-Gln(8)-D-Phe(9 )-Pro(10)). Of these, 4-carboxyphenylalanine (4-Cpa) and phosphotyrosine resulted in high affinity peptides exhibiting K(i) values of 0.85 and 1.1 microM, respectively, 180- and 130-fold increases in potency over the parent cyclic peptide (K(i) = 150 microM). These peptides were noncompetitive with respect to ATP and competitive against the phosphate-accepting substrate, polyGlu(4)Tyr. The truncated cyclic peptide, cyclo(Phe-4-Cpa-Gln-D-Phe-Pro-Asp-Aca) (Aca = epsilon-aminocaproic acid), which did not contain tyrosine, was also a competitive inhibitor with a K(i) value of 24 microM. We conclude that these cyclic peptides bind to a positively charged area that is near the phosphate transfer region of the active site of Src but does not necessarily include the tyrosine-binding pocket. Furthermore, the 4-Cpa-containing cyclic decapeptide shows remarkable selectivity in the inhibition of Src versus the src family members Yes and Lck, as well as other protein tyrosine kinases, Ser/Thr kinases, and other ATP-utilizing enzymes. PMID- 10819991 TI - Tyrosinase and glycoprotein folding: roles of chaperones that recognize glycans. PMID- 10819992 TI - ATP-Dependent histone octamer mobilization and histone deacetylation mediated by the Mi-2 chromatin remodeling complex. AB - The Mi-2 complex has been implicated in chromatin remodeling and transcriptional repression associated with histone deacetylation. Here, we use a purified Mi-2 complex containing six components, Mi-2, Mta 1-like, p66, RbAp48, RPD3, and MBD3, to investigate the capacity of this complex to destabilize histone-DNA interactions and deacetylate core histones. The Mi-2 complex has ATPase activity that is stimulated by nucleosomes but not by free histones or DNA. This nucleosomal ATPase is relatively inefficient, yet is essential to facilitate both translational movement of histone octamers relative to DNA and the efficient deacetylation of the core histones within a mononucleosome. Surprisingly, ATPase activity had no effect on deacetylation of nucleosomal arrays. PMID- 10819993 TI - Electron paramagnetic resonance signals from the S(3) state of the oxygen evolving complex. A broadened radical signal induced by low-temperature near infrared light illumination. AB - The tetranuclear manganese cluster responsible for the oxidation of water in photosystem II cycles through five redox states denoted S(i)() (i = 0, 1, 2, 3, 4). Progress has been made recently in the detection of weak low-field EPR absorptions in both the perpendicular and parallel modes, associated with the integer spin state S(3) [Matsukawa, T., Mino, H., Yoneda, D., and Kawamori, A. (1999) Biochemistry 38, 4072-4077]. We confirm observation of these signals and have obtained them in high yield by illumination of photosystem II membranes, in which the non-heme iron was chemically preoxidized. It is shown that a split g = 4 signal accompanies the S(3) state signals. The signals diminish in the presence of ethanol and vanish in the presence of methanol. This effect is similar to that exerted by these alcohols to the high-spin component (g = 4.1) of the S(2) state and suggests that the latter spin configuration is the precursor of the S(3) state low-field signals. The S(3) state shows similar sensitivity to infrared illumination as has been observed previously in the S(2) state [Boussac, A., Un, S., Horner, O., and Rutherford, A. W. (1998) Biochemistry 37, 4001-4007]. Illumination of the S(3) state with near-infrared light (700-900 nm), at temperatures around 50 K, results in the modification of the low-field signals and most notably to the appearance of a broad (DeltaH approximately 200 G) radical-type signal centered at g = 2. The signal is tentatively assigned to the interaction of the Mn cluster in a modified S(2) state with a radical. PMID- 10819994 TI - Binding of alpha-actinin to titin: implications for Z-disk assembly. AB - Titin is an exceptionally large protein (M.Wt. approximately 3 MDa) that spans half the sarcomere in muscle, from the Z-disk to the M-line. In the Z-disk, it interacts with alpha-actinin homodimers that are a principal component of the Z filaments linking actin filaments. The interaction between titin and alpha actinin involves repeating approximately 45 amino acid sequences (Z-repeats) near the N-terminus of titin and the C-lobe of the C-terminal calmodulin-like domain of alpha-actinin. The conformation of Z-repeat 7 (ZR7) of titin when complexed with the 73-amino acid C-terminal portion of alpha-actinin (EF34) was studied by heteronuclear NMR spectroscopy using (15)N-labeling of ZR7 and found to be helical over a stretch of 18 residues. Complex formation resulted in the protection of one site of preferential cleavage of EF34 at Phe14-Leu17, as determined by limited proteolysis experiments coupled to mass spectrometry measurements. Intermolecular NOEs show Val16 of ZR7 to be positioned close in space to the backbone of EF34 around Phe14. These observations suggest that the mode of binding of ZR7 to EF34 is similar to that of troponin I to troponin C and of peptide C20W to calmodulin. These complexes would appear to represent a general alternative binding mode of calmodulin-like domains to target peptides. PMID- 10819995 TI - ATPase kinetic characterization and single molecule behavior of mutant human kinesin motors defective in microtubule-based motility. AB - Conventional kinesin is a microtubule-based motor protein that is an important model system for understanding mechanochemical transduction. To identify regions of the kinesin protein that participate in microtubule binding and force production, Woehlke et al. [(1997) Cell 90, 207-216] generated 35 alanine mutations in solvent-exposed residues. Here, we have performed presteady-state kinetic and single molecule motility analyses on three of these mutants [Y138A, loop 11 triple (L248A/D249A/E250A), and E311A] that exhibited a similar approximately 3-fold reduction in both microtubule gliding velocity and microtubule-stimulated ATPase activity. All mutants showed normal second-order ATP binding kinetics, indicating correct folding of the active site. The Y138A and loop 11 triple mutants were defective both in nucleotide hydrolysis and in microtubule-stimulated ADP release rates, the latter suggesting a defect in allosteric communication between the microtubule and the active site. A single molecule fluorescence assay further revealed that the loop 11 mutant is defective in initiating processive motion, suggesting that this loop is important for the initial contact between kinesin and the microtubule. Y138A, on the other hand, can bind to the microtubule normally but cannot move processively. For E311A, neither the rate of nucleotide hydrolysis nor ADP release could account for its slower ATPase and gliding velocity, which suggests that either phosphate release or a conformational transition is rate-limiting in this mutant. The single molecule assay showed that E311A has a reduced velocity of movement, but is not defective in processivity. Thus, while these mutants behave similarly in solution ATPase and multiple motor gliding assays, kinetic and single molecule analyses reveal defects in distinct processes in kinesin's mechanochemical cycle. PMID- 10819996 TI - Calcium regulation of gelsolin and adseverin: a natural test of the helix latch hypothesis. AB - The gelsolin family of actin filament binding proteins have highly homologous structures. Gelsolin and adseverin, also known as scinderin, are the most similar members of this family, with adseverin lacking a C-terminal helix found in gelsolin. This helix has been postulated to serve as a calcium-sensitive latch, keeping gelsolin inactive. To test this hypothesis, we have analyzed the kinetics of severing by gelsolin, adseverin, and a gelsolin truncate which lacks the C terminal latch. We find that the relationship between severing rate and calcium ion concentration differs between gelsolin and adseverin, and suggest that calcium controls one rate-limiting step in the activation of adseverin and two in the activation of gelsolin. In contrast, both proteins are activated equally by protons, and have identical severing kinetics at pHs below 6.3. The temperature sensitivity of severing by adseverin and gelsolin is remarkably different, with gelsolin increasing its severing rate 8-fold per 10 degrees C increase in temperature and adseverin increasing its rate only 2-fold per 10 degrees C increase in temperature. Analysis of the gelsolin construct lacking the C terminal helix demonstrates that this helix is responsible for the regulatory differences between gelsolin and adseverin. These results support the C-terminal latch hypothesis for the calcium ion activation of gelsolin. PMID- 10819997 TI - The 1.9 A resolution crystal structure of phosphono-CheY, an analogue of the active form of the response regulator, CheY. AB - To structurally characterize the activated state of the transiently phosphorylated signal transduction protein CheY, we have constructed an alpha thiophosphonate derivative of the CheY D57C point mutant and determined its three dimensional structure at 1.85 A resolution. We have also characterized this analogue with high-resolution NMR and studied its binding to a peptide derived from FliM, CheY's target component of the flagellar motor. The chemically modified derivative, phosphono-CheY, exhibits many of the chemical properties of phosphorylated wild-type CheY, except that it is indefinitely stable. Electron density for the alpha-thiophosphonate substitution is clear and readily interpretable; omit refinement density at the phosphorus atom is greater than 10sigma. The molecule shows a number of localized conformational changes that are believed to constitute the postphosphorylation activation events. The most obvious of these changes include movement of the side chain of the active site base, Lys 109, and a predominately buried conformation of the side chain of Tyr 106. In addition, there are a number of more subtle changes more distant from the active site involving the alpha4 and alpha5 helices. These results are consistent with our previous structural interpretations of other CheY activation mutants, and with our earlier hypotheses concerning CheY activation through propagation of structural changes away from the active site. PMID- 10819998 TI - Features of F(1)-ATPase catalytic and noncatalytic sites revealed by fluorescence lifetimes and acrylamide quenching of specifically inserted tryptophan residues. AB - Catalytic and noncatalytic nucleotide sites of the F(1) sector of ATP synthase were characterized by tryptophan fluorescence techniques. Seven Trp residues inserted in varied microenvironments in the catalytic sites, and one in the noncatalytic sites, were studied in mutant F(1) enzymes which were otherwise devoid of Trp. Parameters measured were fluorescence lifetimes and dynamic and static quenching by acrylamide in the absence or presence of nucleotide. The results indicated that the solution structures of the mutant enzymes were consistent with reported crystal structures. In enzyme with three empty noncatalytic sites, all sites were relatively inaccessible to acrylamide, indicating a closed conformation. In contrast, when the three catalytic sites were empty, they were relatively and equally accessible to acrylamide, indicating an open conformation. This was the case in the presence or absence of Mg(2+). Residue beta-Trp-331 has been extensively used previously to determine nucleotide binding parameters in F(1). Results here showed that in betaY331W mutant F(1), each of the three beta-Trp-331 residues has an unusually long fluorescence lifetime, confirming that each contributes equally to the overall fluorescence signal. PMID- 10819999 TI - Cooperativity in nicotinamide adenine dinucleotide binding induced by mutations of arginine 475 located at the subunit interface in the human liver mitochondrial class 2 aldehyde dehydrogenase. AB - The low-activity Oriental variant of human mitochondrial aldehyde dehydrogenase possesses a lysine rather than a glutamate at residue 487 in the 500 amino acid homotetrameric enzyme. The glutamate at position 487 formed two salt bonds, one to an arginine at position 264 in the same subunit and the other to arginine 475 in a different subunit [Steinmetz, C. G., Xie, P.-G.,Weiner, H., and Hurley, T. D. (1997) Structure 5, 2487-2505]. Mutating arginine 264 to glutamine produced a recombinantly expressed enzyme with nativelike properties; in contrast, mutating arginine 475 to glutamine produced an enzyme that exhibited positive cooperativity in NAD binding. The K(M) for NAD increased 23-fold with a Hill coefficient of 1.8. The binding of both NAD and NADH was affected by the mutation at position 475. Restoring the salt bonds between residues 487 and either or both 264 and 475 did not restore nativelike properties to the Oriental variant. Further, the R475Q mutant was thermally less stable than the native enzyme, Oriental variant, or other mutants. The presence of NAD restored nativelike stability to the mutant. It is concluded that movement of arginine 475 disrupted salt bonds between it and residues other than the one at 487, which caused the apo-R475Q mutant to have properties typical of an enzyme that exhibits positive cooperativity in substrate binding. Breaking the salt bond between glutamate 487 in the Oriental variant and the two arginine residues cannot be the only reason that this enzyme has altered catalytic properties. PMID- 10820000 TI - Recruitment of a double bond isomerase to serve as a reductive dehalogenase during biodegradation of pentachlorophenol. AB - Tetrachlorohydroquinone dehalogenase catalyzes the replacement of chlorine atoms on tetrachlorohydroquinone and trichlorohydroquinone with hydrogen atoms during the biodegradation of pentachlorophenol by Sphingomonas chlorophenolica. The sequence of the active site region of tetrachlorohydroquinone dehalogenase is very similar to those of the corresponding regions of maleylacetoacetate isomerases, enzymes that catalyze the glutathione-dependent isomerization of a cis double bond in maleylacetoacetate to the trans configuration during the catabolism of phenylalanine and tyrosine. Furthermore, tetrachlorohydroquinone dehalogenase catalyzes the isomerization of maleylacetone (an analogue of maleylacetoacetate) at a rate nearly comparable to that of a bona fide bacterial maleylacetoacetate isomerase. Since maleylacetoacetate isomerase is involved in a common and presumably ancient pathway for catabolism of tyrosine, while tetrachlorohydroquinone dehalogenase catalyzes a more specialized reaction, it is likely that tetrachlorohydroquinone dehalogenase arose from a maleylacetoacetate isomerase. The substrates and overall transformations involved in the dehalogenation and isomerization reactions are strikingly different. This enzyme provides a remarkable example of Nature's ability to recruit an enzyme with a useful structural scaffold and elaborate upon its basic catalytic capabilities to generate a catalyst for a newly needed reaction. PMID- 10820001 TI - Structure-based design guides the improved efficacy of deacylation transition state analogue inhibitors of TEM-1 beta-Lactamase(,). AB - Transition state analogue boronic acid inhibitors mimicking the structures and interactions of good penicillin substrates for the TEM-1 beta-lactamase of Escherchia coli were designed using graphic analyses based on the enzyme's 1.7 A crystallographic structure. The synthesis of two of these transition state analogues, (1R)-1-phenylacetamido-2-(3-carboxyphenyl)ethylboronic acid (1) and (1R)-1-acetamido-2-(3-carboxy-2-hydroxyphenyl)ethylboronic acid (2), is reported. Kinetic measurements show that, as designed, compounds 1 and 2 are highly effective deacylation transition state analogue inhibitors of TEM-1 beta lactamase, with inhibition constants of 5.9 and 13 nM, respectively. These values identify them as among the most potent competitive inhibitors yet reported for a beta-lactamase. The best inhibitor of the current series was (1R)-1 phenylacetamido-2-(3-carboxyphenyl)ethylboronic acid (1, K(I) = 5.9 nM), which resembles most closely the best known substrate of TEM-1, benzylpenicillin (penicillin G). The high-resolution crystallographic structures of these two inhibitors covalently bound to TEM-1 are also described. In addition to verifying the design features, these two structures show interesting and unanticipated changes in the active site area, including strong hydrogen bond formation, water displacement, and rearrangement of side chains. The structures provide new insights into the further design of this potent class of beta-lactamase inhibitors. PMID- 10820002 TI - Mapping the functional surface of domain 2 in the gelsolin superfamily. AB - The crystal structure of the F-actin binding domain 2 of severin, the gelsolin homologue from Dictyostelium discoideum, has been determined by multiple isomorphous replacement and refined to 1.75 A resolution. The structure reveals an alpha-helix-beta-sheet sandwich similar to the domains of gelsolin and villin, and contains two cation-binding sites, as observed in other domain 1 and domain 2 homologues. Comparison of the structures of several gelsolin family domains has identified residues that may mediate F-actin binding in gelsolin domain 2 homologues. To assess the involvement of these residues in F-actin binding, three mutants of human gelsolin domain 2 were assayed for F-actin binding activity and thermodynamic stability. Two of the mutants, RRV168AAA and RLK210AAA, demonstrated a lowered affinity for F-actin, indicating a role for those residues in filament binding. Using both structural and biochemical data, we have constructed a model of the gelsolin domain 1-domain 2-F-actin complex. This model highlights a number of interactions that may serve as positive and negative determinants of filament end- and side-binding. PMID- 10820003 TI - Mapping the CD4 binding domain of gp17, a glycoprotein secreted from seminal vesicles and breast carcinomas. AB - gp17, a secretory CD4-binding factor isolated from the human seminal plasma, is identical to the gross cystic disease fluid protein-15, a specific marker for primary and metastatic breast tumors. We previously demonstrated that gp17 binds to CD4 with high affinity and strongly inhibits T lymphocyte apoptosis induced by sequential cross-linking of CD4 and T cell receptor (TCR). To further characterize the gp17/CD4 interaction and map the gp17 binding site, we produced a secreted form of recombinant gp17 fused to human IgG1 Fc, gp17-Ig. We showed that gp17-Ig exhibits a binding affinity for CD4 similar to that of native gp17. As no information about gp17 structure is presently available, 99 overlapping gp17 peptides were synthesized by the Spot method, which allowed the mapping of two CD4 binding regions. Alanine scanning of CD4-reactive peptides identified critical residues, selected for site-directed mutagenesis. Nine gp17-Ig mutants were generated and characterized. Three residues within the carboxy-terminal region were identified as the major binding domain to CD4. The Spot method combined with mutagenesis represents a refined approach to distinguish the contact residues from the ones contributing to the conformation of the CD4 binding domain. PMID- 10820004 TI - Alternative splicing of Wilms' tumor suppressor protein modulates DNA binding activity through isoform-specific DNA-induced conformational changes. AB - The Wilms' tumor suppressor protein (WT1) is a zinc finger transcription factor that appears to function differently according to the presence of a posttranscriptional modification that adds three amino acids into one of the linker regions between the zinc fingers. We have investigated the structural consequences of the insertion of the Lys-Thr-Ser (KTS) sequence by preparing recombinant protein constructs of the four zinc finger DNA-binding domain of WT1 corresponding to the two isoforms with (+KTS) and without (-KTS) the insertion, which is located in the linker region between the third and fourth zinc fingers. NMR resonance assignments were used to estimate the structural differences between the two isoforms both free in solution and in complex with a 14 base pair DNA duplex corresponding to the WT1 recognition element. The NMR spectra indicate that the two isoforms are nearly identical in structure in the absence of the DNA. Only the immediate region of the insertion showed any change in chemical shifts. Upon DNA binding, the NMR spectrum of each isoform changed to indicate greater structure formation in the linker regions. Significant differences were observed between the spectra of the DNA complexes of the +KTS and -KTS isoforms, with the -KTS construct forming a more stable complex, consistent with prior biochemical assays. The majority of the differences between the spectra of the two complexes occur in the immediate region of the insertion, which appears to be closer in structure to the free form of the protein in the case of the +KTS complex. The insertion of the KTS sequence disrupts important interactions of the linker region with the adjacent zinc fingers, thus lowering the stability of the complex. The "normal" (-KTS) sequence of the linker appears to be involved in a C terminal helix-capping interaction with the helix of the preceding zinc finger, a stabilizing interaction which is abrogated in the +KTS isoform. PMID- 10820005 TI - Identification of a binding site for quaternary amines in factor Xa. AB - In the process of characterizing the Na(+)-binding properties of factor Xa, a specific inhibition of this enzyme by quaternary amines was identified, consistent with previous observations. The binding occurs with K(i) in the low millimolar range, with trimethylphenylammonium (TMPA) showing the highest specificity. Binding of TMPA inhibits substrate hydrolysis in a competitive manner, does not inhibit the binding of p-aminobenzamidine to the S1 pocket, and is positively linked to Na(+) binding. Inhibition by TMPA is also seen in thrombin and tissue plasminogen activator (tPA), though to a lesser extent compared to factor Xa. Computer modeling using the crystal structure of factor Xa suggests that TMPA binds to the S2/S3 specificity sites, with its hydrophobic moiety making van der Waals interactions with the side chains of Y99, F174, and W215, and the charged amine coupling electrostatically with the carboxylates of E97. Site-directed mutagenesis of factor Xa, thrombin, and tPA confirms the predictions drawn by docking calculations and reveal a dominant role for residue Y99. Binding of TMPA to factor Xa is drastically (25-fold) reduced by the Y99T replacement. Likewise, the Y99L substitution compromises binding of TMPA to tPA. On the other hand, the affinity of TMPA is enhanced 4-fold in thrombin with the substitution L99Y. The identification of a binding site for quaternary amines in factor Xa has a bearing on the rational design of selective inhibitors of this clotting enzyme. PMID- 10820006 TI - Utilization of site-directed spin labeling and high-resolution heteronuclear nuclear magnetic resonance for global fold determination of large proteins with limited nuclear overhauser effect data. AB - To test whether distances derived from paramagnetic broadening of (15)N heteronuclear single quantum coherence (HSQC) resonances could be used to determine the global fold of a large, perdeuterated protein, we used site directed spin-labeling of 5 amino acids on the surface of (15)N-labeled eukaryotic translation initiation factor 4E (eIF4E). eIF4E is a 25 kDa translation initiation protein, whose solution structure was previously solved in a 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate hydrate (CHAPS) micelle of total molecular mass approximately 45-50 kDa. Distance-dependent line broadening consistent with the three-dimensional structure of eIF4E was observed for all spin-label substitutions. The paramagnetic broadening effects (PBEs) were converted into distances for modeling by a simple method comparing peak heights in (15)N-HSQC spectra before and after reduction of the nitroxide spin label with ascorbic acid. The PBEs, in combination with HN-HN nuclear Overhauser effects (NOEs) and chemical shift index (CSI) angle restraints, correctly determined the global fold of eIF4E with a backbone precision of 2.3 A (1.7 A for secondary structure elements). The global fold was not correctly determined with the HN-HN NOEs and CSI angles alone. The combination of PBEs with simulated restraints from another nuclear magnetic resonance (NMR) method for global fold determination of large proteins (methyl-protonated, highly deuterated samples) improved the quality of calculated structures. In addition, the combination of the two methods simulated from a crystal structure of an all alpha-helical protein (40 kDa farnesyl diphoshphate synthase) correctly determined the global fold where neither method individually was successful. These results show the potential feasibility of obtaining medium-resolution structures for proteins in the 40-100 kDa range via NMR. PMID- 10820007 TI - Snapping of the carboxyl terminal tail of the catalytic subunit of PKA onto its core: characterization of the sites by mutagenesis. AB - A set of 45 mutants of the carboxyl terminal tail of the PKA catalytic subunit was prepared and used to assess the contribution of this tail to the structure and function of the kinase. Ala substitutions of Asp 323, Phe 327, Glu 333, and Phe 350 resulted in a complete loss of enzymatic activity. Other replacements by Ala (Phe 314, Tyr 330, Glu 332, and Phe 347) brought about either a drop in activity to less than 10% of the wild-type enzyme or a reduction of affinity toward ATP (Lys 317, Lys 319, Tyr 330, and Glu 332) or toward Kemptide (Ile 315, Tyr 330, Val 337, Ile 339, Lys 345, and Glu 346). Mutations of Ser 338, a major autophosphorylation site of PKA, by Ala, Glu, Asp, Gln, and Asn showed that the kinetic parameters of these mutants are similar to those of the wild-type. The contribution of each of these tail mutations to the structure and stability of the kinase was assessed by monitoring its effect on the heat stability (when measurable) or by determining the susceptibility of the mutant kinase to cleavage by the Kinase Splitting Membranal Proteinase/Meprin beta. Here we show that the tail of PKA has a key role in creating the active conformation of the kinase. It does so by means of specific amino acid residues, which act as "snapping points" to embrace the two lobes of the kinase and orient them in the correct juxtaposition for substrate docking, biorecognition, and catalysis. PMID- 10820008 TI - Deimination of myelin basic protein. 1. Effect of deimination of arginyl residues of myelin basic protein on its structure and susceptibility to digestion by cathepsin D. AB - The effect of deimination of arginyl residues in bovine myelin basic protein (MBP) on its susceptibility to digestion by cathepsin D has been studied. Using bovine component 1 (C-1) of MBP, the most unmodified of the components with all 18 arginyl residues intact, we have generated a number of citrullinated forms by treatment of the protein with purified peptidylarginine deiminase (PAD) in vitro. We obtained species containing 0-9.9 mol of citrulline/mol of MBP. These various species were digested with cathepsin D, a metalloproteinase which cleaves proteins at Phe-Phe linkages. The rate of digestion compared to component 1 was only slightly affected when 2.7 or 3.8 mol of citrulline/mol of MBP was present. With 7.0 mol of citrulline/mol of MBP, a large increase in the rate of digestion occurred. No further increase was observed with 9.9 mol of citrulline/mol of MBP. The immunodominant peptide 43-88 (bovine sequence) was released slowly when 2.7 and 3.8 mol of citrulline/mol of MBP was present, but it was released rapidly when 7.0 mol of citrulline/mol of MBP was present. The dramatic change in digestion with 7.0 mol of citrulline/mol of MBP or more could be explained by a change in three-dimensional structure. Molecular dynamics simulation showed that increasing the number of citrullinyl residues above 7 mol/mol of MBP generated a more open structure, consistent with experimental observations in the literature. We conclude that PAD, which deiminates arginyl residues in proteins, decreases both the charge and compact structure of MBP. This structural change allows better access of the Phe-Phe linkages to cathepsin D. This scheme represents an effective way of generating the immunodominant peptide which sensitizes T-cells for the autoimmune response in demyelinating disease. PMID- 10820009 TI - Deimination of myelin basic protein. 2. Effect of methylation of MBP on its deimination by peptidylarginine deiminase. AB - Deimination of myelin basic protein (MBP) has been implicated in the chemical pathogenesis of multiple sclerosis (MS). Degradation of bovine MBP by cathepsin D, a myelin-associated protease, was increased when 6 arginyl residues were deiminated and became very rapid when all 18 arginyl residues were deiminated. Since MBP contains a number of modifications, including methylation, phosphorylation, etc., we studied the effect of methylation, an irreversible modification, to determine how this modification affected deimination. Methylation of Arg 106 in bovine MBP (Arg 107 in human), a naturally occurring modification of MBP, has been shown to affect the deimination of arginyl residues in the present study. Since fractionation of MBP into unmethylated, monomethylated, and dimethylated species cannot be done readily on a preparative scale, mass spectrometry with the Q-TOF instrument resolved these species readily since each differed from the other by 14 atomic mass units (amu). Examination of five different hMBP samples, two from normal brain and three from MS brain, revealed that increased deimination of arginyl residues correlated with a decreased methylation of Arg 107 (human sequence). To study this process in vitro, bovine MBP (bMBP) was used. Component 1 (C-1) is the most cationic of the MBP "charge isomers" and the most unmodified, in which all arginyl residues are intact. It was deiminated to various extents with purified bovine brain peptidylarginine deiminase, generating a number of species containing 0-13.7 mol of citrulline/mol of bMBP. Mass spectrometry of each of these species permitted us to determine the influence of methylation of Arg 106 (bovine sequence) on deimination by this enzyme. We found that bMBP with unmethylated arginine was deiminated at a rate of 0.081 mol of citrulline/min, with monomethylarginine, 0.068 mol of citrulline/min, and with dimethylarginine, 0.036 mol of citrulline/min. We suggest that the methylated arginyl residue becomes sequestered in the hydrophobic beta-sheet structure and disrupts the three dimensional structure of the protein so that other arginyl residues are less accessible to peptidylarginine deiminase. PMID- 10820010 TI - Fluoride inhibition of Klebsiella aerogenes urease: mechanistic implications of a pseudo-uncompetitive, slow-binding inhibitor. AB - Klebsiella aerogenes urease uses a dinuclear nickel active site to catalyze the hydrolysis of urea. Here, we describe the steady-state and pre-steady-state kinetics of urease inhibition by fluoride. Urease is slowly inhibited by fluoride in both the presence and absence of substrate. Steady-state rate studies yield parallel double-reciprocal plots; however, we show that fluoride interaction with urease is not compatible with classical uncompetitive inhibition. Rather, we propose that fluoride binds to an enzyme state (E) that is in equilibrium with resting enzyme (E) and produced during catalysis. Fluoride binding rates are directly proportional to inhibitor concentration. Substrate reduces both the rate of fluoride binding to urease and the rate of fluoride dissociation from the complex, consistent with urea binding to E and E.F in addition to E. Fluoride inhibition is pH-dependent due to a protonation event linked to fluoride dissociation. Fluoride binding is pH-independent, suggesting that fluoride anion, not HF, is the actual inhibitor. We assess the kinetic results in terms of the known protein crystal structure and evaluate possible molecular interpretations for the structure of the E state, the site of fluoride binding, and the factors associated with fluoride release. Finally, we note that the apparent uncompetitive inhibition by fluoride as reported for several other metalloenzymes may need to be reinterpreted in terms of fluoride interaction with the corresponding E states. PMID- 10820011 TI - Roles of two conserved amino acid residues in the active site of galactose-1 phosphate uridylyltransferase: an essential serine and a nonessential cysteine. AB - Galactose-1-phosphate uridylyltransferase (GalT) catalyzes the reversible transformation of uridine 5'-diphosphate glucose (UDPGlc) and galactose-1 phosphate into uridine 5'-diphosphate galactose (UDPGal) and glucose-1-phosphate through a double displacement mechanism, with the intermediate formation of a covalent uridylyl-enzyme (UMP-enzyme). The covalent linkage is a phosphoramidate formed between the UMP moiety and the His 166 N(epsilon)(2) of GalT, with His 166 N(delta1) retaining a proton throughout the catalytic cycle. Cys 160 and Ser 161 in Escherichia coli GalT are engaged in hydrogen bonding with the peripheral phosphoryl oxygen atoms of the substrate in the crystalline UMP-enzyme and in the crystalline complex of H166G-GalT with UDPGlc [Wedekind, J. E., Frey, P. A., and Rayment, I. (1996) Biochemistry 35, 11560-11569; Thoden, J. B., Ruzicka, F. J., Frey, P. A., Rayment, I., and Holden, H. M. (1997) Biochemistry 36, 1212-1222]. Site-directed mutagenesis, thermodynamic, transient kinetic, and steady-state kinetic studies have been performed to investigate the roles of Cys 160 and Ser 161 in catalysis. The absence of the thiol group of Cys 160 in the variants C160S and C160A did not seriously alter the enzymatic activity. However, the variant S161A displayed 7000-fold less activity than wild-type GalT. The low activity of S161A was directly related to impaired uridylylation rate constant (3.7 x 10( )(2) s(-)(1)) and de-uridylylation rate constant (0.5 x 10(-)(2) s(-)(1)) resulting from a higher kinetic barrier for uridylyl-group transfer by the variant S161A as compared with the wild-type GalT. Equilibrium uridylylation studies showed that neither Cys 160 nor Ser 161 was involved in stabilizing the uridylyl-enzyme intermediate. The results lead to the conclusion that the conserved Cys 160 does not play a critical role in catalysis. Ser 161 is most likely involved in donating a hydrogen bond to the beta-phosphoryl group of a substrate, thereby providing proper orientation for nucleophilic catalysis. PMID- 10820012 TI - Involvement of a novel hydroxylamine oxidoreductase in anaerobic ammonium oxidation. AB - In this study a novel hydroxylamine oxidoreductase (HAO) was purified and characterized from an anaerobic ammonium-oxidizing (Anammox) enrichment culture. The enzyme, which constituted about 9% of the protein mass in the soluble fraction of the cell extract, was able to oxidize hydroxylamine and hydrazine. When phenazine methosulfate and methylthiazolyltetrazolium bromide were used as electron acceptors, a V(max) [21 and 1.1 micromol min(-)(1) (mg of protein)( )(1)] and K(m) (26 and 18 microM) for hydroxylamine and hydrazine were determined, respectively. The hydroxylamine oxidoreductase is a trimer and contains about 26 hemes per 183 kDa. As deduced from UV/vis spectra, hydroxylamine reduced more and different cytochromes than hydrazine. The dithionite-reduced spectrum showed an unusual 468 nm peak. Inhibition experiments with H(2)O(2) showed that hydroxylamine bound to this P-468 cytochrome, which is assumed to be the putative substrate binding site. Cyanide and hydrazine inhibited the oxidation of hydroxylamine. The amino acid sequences of several peptide fragments of HAO from Anammox showed a clear difference with the deduced amino acid sequence of HAO from the aerobic ammonia-oxidizing bacterium Nitrosomonas europaea. In EPR spectra of the Anammox HAO, two g-values (g(z)() = 2.37 and 2.42) were observed, which were not present in HAO of N. europaea. PMID- 10820013 TI - Cobalt(2+) binding to human and tomato copper chaperone for superoxide dismutase: implications for the metal ion transfer mechanism. AB - The copper chaperone for superoxide dismutase (CCS) gene encodes a protein that is believed to deliver copper ions specifically to copper-zinc superoxide dismutase (CuZnSOD). CCS proteins from different organisms share high sequence homology and consist of three distinct domains; a CuZnSOD-like central domain 2 flanked by domains 1 and 3, which contain putative metal-binding motifs. We report deduced protein sequences from tomato and Arabidopsis, the first functional homologues of CCS identified in plants. We have purified recombinant human (hCCS) and tomato (tCCS) copper chaperone proteins, as well as a truncated version of tCCS containing only domains 2 and 3. Their cobalt(2+) binding properties in the presence and absence of mercury(2+) were characterized by UV vis and circular dichroism spectroscopies and it was shown that hCCS has the ability to bind two spectroscopically distinct cobalt ions whereas tCCS binds only one. The cobalt binding site that is common to both hCCS and tCCS displayed spectroscopic characteristics of cobalt(2+) bound to four or three cysteine ligands. There are only four cysteine residues in tCCS, two in domain 1 and two in domain 3; all four are conserved in other CCS sequences including hCCS. Thus, an interaction between domain 1 and domain 3 is concluded, and it may be important in the copper chaperone mechanism of these proteins. PMID- 10820014 TI - Substitution of tyrosine for the proximal histidine ligand to the heme of prostaglandin endoperoxide synthase 2: implications for the mechanism of cyclooxygenase activation and catalysis. AB - Prostaglandin H(2) synthesis by prostaglandin endoperoxide synthase (PGHS) requires the heme-dependent activation of the protein's cyclooxygenase activity. The PGHS heme participates in cyclooxygenase activation by accepting an electron from Tyr385 located in the cyclooxygenase active site. Two mechanisms have been proposed for the oxidation of Tyr385 by the heme iron: (1) ferric enzyme oxidizes a hydroperoxide activator and the incipient peroxyl radical oxidizes Tyr385, or (2) ferric enzyme reduces a hydroperoxide activator and the incipient ferryl-oxo heme oxidizes Tyr385. The participation of ferrous PGHS in cyclooxygenase activation was evaluated by determining the reduction potential of PGHS-2. Under all conditions tested, this potential (<-135 mV) was well below that required for reactions leading to cyclooxygenase activation. Substitution of the proximal heme ligand, His388, with tyrosine was used as a mechanistic probe of cyclooxygenase activation. His388Tyr PGHS-2, expressed in insect cells and purified to homogeneity, retained cyclooxygenase activity but its peroxidase activity was diminished more than 300-fold. Concordant with this poor peroxidase activity, an extensive lag in His388Tyr cyclooxygenase activity was observed. Addition of hydroperoxides resulted in a concentration-dependent decrease in lag time consistent with each peroxide's ability to act as a His388Tyr peroxidase substrate. However, hydroperoxide treatment had no effect on the maximal rate of arachidonate oxygenation. These data imply that the ferryl-oxo intermediates of peroxidase catalysis, but not the Fe(III)/Fe(II) couple of PGHS, are essential for cyclooxygenase activation. In addition, our findings are strongly supportive of a branched-chain mechanism of cyclooxygenase catalysis in which one activation event leads to many cyclooxygenase turnovers. PMID- 10820015 TI - The role of lysine 529, a conserved residue of the acyl-adenylate-forming enzyme superfamily, in firefly luciferase. AB - Firefly luciferase catalyzes the highly efficient emission of yellow-green light from the substrates luciferin, Mg-ATP, and oxygen in a two-step process. The enzyme first catalyzes the adenylation of the carboxylate substrate luciferin with Mg-ATP followed by the oxidation of the acyl-adenylate to the light-emitting oxyluciferin product. The beetle luciferases are members of a large family of nonbioluminescent proteins that catalyze reactions of ATP with carboxylate substrates to form acyl-adenylates. Formation of the luciferase-luciferyl-AMP complex is a specific example of the chemistry common to this enzyme family. Site directed mutants at positions Lys529, Thr343, and His245 were studied to determine the effects of the amino acid changes at these positions on the individual luciferase-catalyzed adenylation and oxidation reactions. The results suggest that Lys529 is a critical residue for effective substrate orientation and that it provides favorable polar interactions important for transition state stabilization leading to efficient adenylate production. These findings as well as those with the Thr343 and His245 mutants are interpreted in the context of the firefly luciferase X-ray structures and computational-based models of the active site. PMID- 10820016 TI - Role of the kidney in regulating the metabolism of HDL in rabbits: evidence that iodination alters the catabolism of apolipoprotein A-I by the kidney. AB - To evaluate the factors that regulate HDL catabolism in vivo, we have measured the clearance of human apoA-I from rabbit plasma by following the isotopic decay of (125)I-apoA-I and the clearance of unlabeled apoA-I using a radioimmunometric assay (RIA). We show that the clearance of unlabeled apoA-I is 3-fold slower than that of (125)I-apoA-I. The mass clearance of iodinated apoA-I, as determined by RIA, is superimposable with the isotopic clearance of (125)I-apoA-I. The data demonstrate that iodination of tyrosine residues alters the apoA-I molecule in a manner that promotes an accelerated catabolism. The clearance from rabbit plasma of unmodified apoA-I on HDL(3) and a reconstituted HDL particle (LpA-I) were very similar and about 3-4-fold slower than that for (125)I-apoA-I on the lipoproteins. Therefore, HDL turnover in the rabbit is much slower than that estimated from tracer kinetic studies. To determine the role of the kidney in HDL metabolism, the kinetics of unmodified apoA-I and LpA-I were reevaluated in animals after a unilateral nephrectomy. Removal of one kidney was associated with a 40-50% reduction in creatinine clearance rates and a 34% decrease in the clearance rate of unlabeled apoA-I and LpA-I particles. In contrast, the clearance of (125)I-labeled molecules was much less affected by the removal of a kidney; FCR for (125)I-LpA-I was reduced by <10%. The data show that the kidneys are responsible for most (70%) of the catabolism of apoA-I and HDL in vivo, while (125)I-labeled apoA-I and HDL are rapidly catabolized by different tissues. Thus, the kidney is the major site for HDL catabolism in vivo. Modification of tyrosine residues on apoA-I may increase its plasma clearance rate by enhancing extra renal degradation pathways. PMID- 10820017 TI - Novel purification scheme and functions for a C3-binding protein from Streptococcus pneumoniae. AB - To isolate microbial proteins capable of binding the third component of complement (C3), we coupled the free sulfhydryl group of methylamine-inactivated C3 to a thiolSepharose matrix. This simple technique facilitated the purification of the first C3-binding protein isolated from a bacterium (Streptococcus pneumoniae). Both metastable (native) and thioester-disrupted C3 were recognized by this protein; binding of C3 was noncovalent, independent of thioester conformation, and preferential for the C3 alpha-chain. Sequencing of amino terminal and internal peptides from the C3-binding protein disclosed a proline rich region spanning approximately 20 amino acids and a signal peptide that had not been previously reported. The gene was isolated from a library of genomic DNA from laboratory strain CP1200 by screening with a 1200 bp PCR product amplified from degenerate oligonucleotides encoding the amino terminal sequence and the internal proline-rich sequence. The open reading frame spanned 1692 bp; all peptide sequences were identified in the translated gene product, which also contained at least three choline-binding repeats at the carboxy-terminus. The gene was conserved, and the translated protein was functionally active in pneumococcal clinical isolates of serotypes 1, 3, 4, 14, and 19F. Serum from a patient recovering from acute pneumococcal infection contained IgG antibodies specific for this protein by immunoblot. Wide conservation among clinical isolates, saturable binding of C3, and the ability to stimulate the human immune response have not previously been reported for this choline-binding protein. A similar biochemical approach should enable the identification of other C3-binding proteins in microorganisms able to elude complement-mediated host defense. PMID- 10820018 TI - Biphasic kinetics of activation and signaling for PAR1 and PAR4 thrombin receptors in platelets. AB - Thrombin activates platelets in an ordered sequence of events that includes shape change, increase in cytoplasmic Ca(2+), activation of the alphaIIbbeta3 integrin, granule secretion, aggregation, and formation of a stable hemostatic plug. Activation of this process has also been implicated in the pathogenesis of atherosclerosis, stroke, and thrombosis. There are two identified thrombin activated receptors on the surface of human platelets. PAR1 is a high-affinity thrombin receptor, and PAR4 is a low apparent affinity thrombin receptor of uncertain function. The goal of these studies is to determine the kinetics of thrombin activation of PAR1 and PAR4 and to relate the individual inputs from each receptor to platelet Ca(2+) signaling, secondary autocrine stimulation, and aggregation. Using a combination of PAR-specific peptide ligands and anti-PAR1 reagents, we separated the biphasic thrombin Ca(2+) response of platelets into two discrete components-a rapid spike response caused by PAR1, followed by a slower prolonged response from PAR4. Despite having a 20-70-fold slower rate of activation, PAR4 produces the majority of the integrated Ca(2+) signal that is sustained by the continuous presence of catalytically active thrombin. Surprisingly, PAR4 activation is much more effective than PAR1 activation in mounting secondary autocrine Ca(2+) signals from secreted ADP. The strong ADP response due to activated PAR4, however, requires prior activation of PAR1 as would normally occur during treatment of platelets with thrombin. Thus, the late signal generated by activated PAR4 is not redundant with the early signal from PAR1 and instead serves to greatly extend the high intracellular Ca(2+) levels that support the late phase of the platelet aggregation process. PMID- 10820019 TI - Self-association of human protein S. AB - Protein S functions as a cofactor with activated protein C in the down-regulation of the blood coagulation cascade. In vitro studies have historically produced conflicting data with regard to the extent of various protein S activity in clotting assays which typically involve adding CaCl(2) to initiate reactions. We report here that protein S reversibly self-associates in the absence of Ca(2+). Sedimentation experiments showed a transition in sedimentation velocity from 7.2 to 4.2 S with a transition midpoint (T(m)) of 0.42 mM Ca(2+) for intact protein S. Studies of thrombin cleaved (Arg(70)) protein S revealed similar results with a transition in sedimentation velocity from 7.9 to 4.4 S with a T(m) of 0.42 mM Ca(2+). This transition is reversible with the addition of 10 mM EDTA. Sedimentation equilibrium data suggest at a minimum, a monomer-dimer-trimer association. Sedimentation velocity experiments were also performed on mixtures of protein S and prothrombin which showed no heterodimer formation in either Ca(2+) or EDTA solutions. These data suggest that previous interpretations of protein S structure and function may have been confounded by the self-associative behavior of protein S in non-Ca(2+) solutions. PMID- 10820020 TI - Activated leukocytes oxidatively damage DNA, RNA, and the nucleotide pool through halide-dependent formation of hydroxyl radical. AB - A variety of chronic inflammatory conditions are associated with an increased risk for the development of cancer. Because of the numerous links between DNA oxidative damage and carcinogenesis, a potential role for leukocyte-generated oxidants in these processes has been suggested. In the present study, we demonstrate a novel free transition metal ion-independent mechanism for hydroxyl radical ((*)OH)-mediated damage of cellular DNA, RNA, and cytosolic nucleotides by activated neutrophils and eosinophils. The mechanism involves reaction of peroxidase-generated hypohalous acid (HOCl or HOBr) with intracellular superoxide (O(2)(*)(-)) forming (*)OH, a reactive oxidant species implicated in carcinogenesis. Incubation of DNA with either isolated myeloperoxidase (MPO) or eosinophil peroxidase (EPO), plasma levels of halides (Cl(-) and Br(-)), and a cell-free O(2)(*)(-) -generating system resulted in DNA oxidative damage. Formation of 8-hydroxyguanine (8-OHG), a mutagenic base which is a marker for (*)OH-mediated DNA damage, required peroxidase and halides and occurred in the presence of transition metal chelators (DTPA +/- desferrioxamine), and was inhibited by catalase, superoxide dismutase (SOD), and scavengers of hypohalous acids. Similarly, exposure of DNA to either neutrophils or eosinophils activated in media containing metal ion chelators resulted in 8-OHG formation through a pathway that was blocked by peroxidase inhibitors, hypohalous acid scavengers, and catalytically active (but not heat-inactivated) catalase and SOD. Formation of 8-OHG in target cells (HA1 fibroblasts) occurred in all guanyl nucleotide containing pools examined following exposure to both a low continuous flux of HOCl (at sublethal doses, as assessed by [(14)C]adenine release and clonogenic survival), and hyperoxia (to enhance intracellular O(2)(*)(-) levels). Mitochondrial DNA, poly A RNA, and the cytosolic nucleotide pool were the primary targets for oxidation. Moreover, modest but statistically significant increases in the 8-OHG content of nuclear DNA were also noted. These results suggest that the peroxidase-H(2)O(2)-halide system of leukocytes is a potential mechanism contributing to the well-established link between chronic inflammation, DNA damage, and cancer development. PMID- 10820021 TI - DNA-Bound peptide radicals generated through DNA-mediated electron transport. AB - Flash-quench experiments were carried out to explore peptide/DNA electron transfer reactions. DNA-bound [Ru(phen)(2)(dppz)](3+) (phen = 1,10 phenanthroline; dppz = dipyridophenazine) and [Ru(phen)(bpy')(dppz)](3+) [bpy' = 4-(4'-methyl-2, 2'-bipyridyl)valerate], generated in situ by flash-quench methodology, are powerful ground-state oxidants, capable of oxidizing guanine or tyrosine intercalated in DNA. In flash-quench experiments with mixed-sequence oligonucleotides in the presence of Lys-Tyr-Lys, transient absorption spectroscopy yielded a spectrum with a sharp maximum at 405 nm assigned to the tyrosine radical. Experiments with poly(dG.dC) suggested the intermediacy of the guanine radical, since the rise of the 405 nm signal occurred with the same kinetics as the disappearance of the guanine radical, as monitored at 510 nm. In oligonucleotide duplexes containing [Ru(phen)(bpy')(dppz)](2+) tethered at one end, damage to distant guanines was observed by gel electrophoresis, consistent with the mobility of the electron hole through the DNA duplex; the presence of the peptide did not inhibit but instead altered the distribution of guanine damage. Covalent adducts of the DNA and Lys-Tyr-Lys were detected as final irreversible products of this peptide-to-DNA electron-transfer chemistry by mass spectrometric and enzymatic digestive analysis. From these different assays and comparison of reactions of Lys-Trp-Lys and Lys-Tyr-Lys, the reactivity of the DNA bound tyrosine radical was found to differ considerably from that of the tryptophan radical. These results establish that Lys-Tyr-Lys and Lys-Trp-Lys can participate in long-range electron-transfer reactions through the DNA from a distinct binding site. On that basis, proposals for functional roles for these peptide radicals may be considered. PMID- 10820022 TI - Role for the C-terminus in agonist-induced mu opioid receptor phosphorylation and desensitization. AB - Determining which domains and amino acid residues of the mu opioid receptor are phosphorylated is critical for understanding the mechanism of mu opioid receptor phosphorylation. The role of the C-terminus of the receptor was investigated by examining the C-terminally truncated or point-mutated mu opioid receptors in receptor phosphorylation and desensitization. Both wild-type and mutated receptors were stably expressed in Chinese hamster ovary (CHO) cells. The receptor expression was confirmed by receptor radioligand binding and immunoblottting. After exposure to 5 microM of DAMGO, phosphorylation of the C terminally truncated receptor and the mutant receptor T394A was reduced to 40 and 10% of that of the wild-type receptor, respectively. Mutation effects on agonist induced desensitization were studied using adenylyl cyclase inhibition assays. The C-terminally truncated receptor and mutant receptor T394A both showed complete loss of DAMGO-induced desensitization, while the mutant T/S-7A receptor only lost part of its ability to desensitize. Taken together, these results suggest that the C-terminus of the mu opioid receptor participates in receptor phosphorylation and desensitization with threonine 394, a crucial residue for both features. DAMGO-induced mu opioid receptor phosphorylation and desensitization are associated and appear to involve both the mu opioid receptor C-terminus and other domains of the receptor. PMID- 10820023 TI - Synthesis and characterization of photolabile derivatives of serotonin for chemical kinetic investigations of the serotonin 5-HT(3) receptor. AB - A series of photolabile o-nitrobenzyl derivatives of serotonin (caged serotonin) were synthesized: the amine-linked serotonin derivatives N-(2-nitrobenzyl) serotonin (Bz-5HT) and N-(alpha-carboxy-2-nitrobenzyl) serotonin (N-CNB-5HT), and O-alpha-carboxy-2-nitrobenzyl) serotonin (O-CNB-5HT), which has the caging group attached to the phenolic OH group. All the derivatives released free serotonin when excited by 308-nm or 337-nm laser pulses. The time constant of serotonin release from N-CNB-5HT was 1. 2 ms, with a quantum yield of 0.08. This is too slow for rapid chemical kinetic measurements. O-CNB-5HT is suitable for transient kinetic investigations of the serotonin 5-HT(3) receptor. It released serotonin with a time constant of 16 micros and a quantum yield of 0.03. The biological properties of O-CNB-5HT were evaluated, and the applicability of the compound for kinetic studies of the 5-HT(3) receptor was demonstrated. O-CNB-5HT does not activate the 5-HT(3) receptor by itself, nor does it modulate the response of a cell when co-applied with serotonin. When irradiated with a 337-nm laser pulse, O CNB-5HT released free serotonin that evoked 5-HT(3) receptor-mediated whole-cell currents in NIE-115 mouse neuroblastoma cells. PMID- 10820024 TI - Posttranslational modifications of the lutropin receptor: mass spectrometric analysis. AB - Our present knowledge of the lutropin (LH/hCG) receptor structure derives from deductions made from its amino acid sequence as established by studying the cDNA. To obtain direct experimental information, luteinizing hormone (LH) receptor expressed in L cells was immunopurified in sufficient amounts to warrant analysis by mass spectrometry and microsequencing. The mature receptor, complexed to human chorionic gonadotropin (hCG), was purified by using monoclonal antibodies recognizing the hormone, whereas the mannose-rich non-hormone-binding precursor was purified by use of antireceptor antibodies. Determination of the N-terminus showed that (2)/(3) of protein molecules started at Thr24 whereas (1)/(3) started at Ala28. All these molecules bound hCG, suggesting that the most N-terminal region of the receptor does not participate in hormone binding. Six N glycosylation sites have been predicted from the amino acid sequence. One of them (Asn299) was found to be nonglycosylated in both the precursor and the mature protein. The most heavily glycosylated residue was Asn291, followed by Asn195 and Asn99. These three sites accounted for 82% and 97% of carbohydrate moieties in the mature receptor and in the mannose-rich precursor, respectively. The presence of some receptor molecules nonglycosylated at sites 99, 174, and 195 in hormone receptor complexes dismisses a direct role of these glycosylation sites in hormone binding or in the correct folding of the protein. The mature carbohydrate chains were homogeneous at position 174, 195, and 313 (absence of Golgi mannosidase II activity at positions 174 and 313, absence of GlcNAc tranferases III and IV activity at position 195). Heterologous carbohydrates were present at sites 99 and 291. The latter, which is highly variable in carbohydrate chains, is unlikely to participate in a direct interaction with hormone. Site 313 thus remains as the main candidate for a role in hormone binding. PMID- 10820025 TI - Analysis of the properties of the N-terminal nucleotide-binding domain of human P glycoprotein. AB - Human P-glycoprotein, the MDR1 gene product, requires both Mg(2+)-ATP binding and hydrolysis to function as a drug transporter; however, the mechanism(s) defining these events is not understood. In the present study, we explored the nature of Mg(2+)-ATP binding in the N-terminal nucleotide-binding domain of human P glycoprotein and identified the minimal functional unit required for specific ATP binding. Recombinant proteins encompassing amino acids within the region beginning at 348 and ending at 707 were expressed in Escherichia coli, purified from inclusion bodies under denaturing conditions, and renatured by rapid dilution. The ability of ATP to interact with these proteins was examined by use of the photoactive ATP analogue [alpha-(32)P]-8-azido-ATP. Photoaffinity labeling of recombinant proteins identified the region between amino acids 375 and 635 as the region necessary to obtain specific ATP-binding properties. Specific protein labeling was saturable, enhanced by Mg(2+), and inhibited by ATP. Recombinant proteins confined within the region beginning at amino acid 392 and ending at amino acid 590 demonstrated nonspecific [alpha-(32)P]-8-azido-ATP labeling. Nonspecific labeling was not enhanced by Mg(2+) and was inhibited only by high concentrations of ATP. Using a D555N mutated protein, we found that the conserved aspartate residue in the Walker B motif plays a role in magnesium-enhanced ATP binding. Taken together, these data define the region of the N-terminal nucleotide-binding domain of P-glycoprotein that is required for specific ATP binding and suggest that magnesium may play a role in stabilizing the ATP-binding site. PMID- 10820026 TI - Identification of the carbonic anhydrase II binding site in the Cl(-)/HCO(3)(-) anion exchanger AE1. AB - The human Cl(-)/HCO(3)(-) anion exchanger (AE1) possesses a binding site within its 33 residue carboxyl-terminal region (Ct) for carbonic anhydrase II (CAII). The amino acid sequence comprising this CAII binding site was determined by peptide competition and by testing the ability of truncation and point mutants of the Ct sequence to bind CAII with a sensitive microtiter plate binding assay. A synthetic peptide consisting of the entire 33 residues of the Ct (residues 879 911) could compete with a GST fusion protein of the Ct (GST-Ct) for binding to immobilized CAII, while a peptide consisting of the last 16 residues (896-911) could not. A series of truncation mutants of the GST-Ct showed that the terminal 21 residues of AE1 were not required for binding CAII. Removal of four additional residues (887-890) from the Ct resulted in loss of CAII binding. Acidic residues in this region (D887ADD) were critical for binding since mutating this sequence in the GST-Ct to DAAA, AAAA, or NANN caused loss of CAII binding. A GST-Ct construct mutated to D887ANE, the homologous sequence in AE2, could bind CAII. AE2 is a widely expressed anion exchanger and has a homologous Ct region with 60% sequence identity to AE1. A GST fusion protein of the 33 residue Ct of AE2 could bind to CAII similarly to the Ct of AE1. Tethering of CAII to an acidic motif within the Ct of anion exchangers may be a general mechanism for promoting bicarbonate transport across cell membranes. PMID- 10820027 TI - Annexin V binding perturbs the cardiolipin fluidity gradient in isolated mitochondria. Can it affect mitochondrial function? AB - The phosholipid bilayer fluidity of isolated mitochondria and phospholipid vesicles after calcium-dependent binding of annexin V was studied using EPR spectroscopy. The membranes were probed at different depths by alternatively using cardiolipin, phosphatidylcholine, or phosphatidylethanolamine spin labeled at position C-5 or C-12 or C-16 of the beta acyl chain. Computer-aided spectral titration facilitated observing and quantitating the EPR spectrum from phospholipid spin labels affected by annexin binding, and spectral mobility was calibrated by comparison with standard spectra scanned at various temperatures. In most cases it was found that binding of the protein to the membranes makes the inner bilayer more rigid up to acyl position C-12 than afterward, in agreement with the previously observed effect in SUVs [Megli, F. M., Selvaggi, M., Liemann, S., Quagliariello, E., and Huber, R. (1998) Biochemistry 37, 10540-10546]. Moreover, in isolated mitochondrial membranes, cardiolipin apparently is more readily affected than the other main phospholipids, while in vesicles made from mitochondrial phospholipids, the different species are affected in essentially the same way. This behavior is consistent with the existence of distinct cardiolipin pools in mitochondria, and with the already advanced hypothesis that these domains are the binding site for annexin V to the isolated organelles [Megli, F. M., Selvaggi, M., De Lisi, A., and Quagliariello, E. (1995) Biochim. Biophys. Acta 1236, 273-278]. Keeping in mind the funcional importance of cardiolipin in the mitochondrial membrane, the question is raised as to whether the observed influence of annexin V binding to this phospholipid and its consequent local fluidity alteration might affect the mitochondrial functionality, at least in vitro. PMID- 10820028 TI - Nodulisporic acid opens insect glutamate-gated chloride channels: identification of a new high affinity modulator. AB - Nodulisporic acid (NA) is an indole diterpene fungal product with insecticidal activity. NA activates a glutamate-gated chloride channel (GluCl) in grasshopper neurons and potentiates channel opening by glutamate. The endectocide ivermectin (IVM) induces a similar, but larger current than NA. Using Drosophila melanogaster head membranes, a high affinity binding site for NA was identified. Equilibrium binding studies show that an amide analogue, N-(2-hydroxyethyl-2,2 (3)H)nodulisporamide ([(3)H]NAmide), binds to a single population of sites in head membranes with a K(D) of 12 pM and a B(max) of 1.4 pmol/mg of protein. A similar K(D) is determined from the kinetics of ligand binding and dissociation. Four lines of evidence indicate that the binding site is a GluCl. First, NA potentiates opening of a glutamate-gated chloride current in grasshopper neurons. Second, glutamate inhibits the binding of [(3)H]NAmide by increasing the rate of dissociation 3-fold. Third, IVM potently inhibits the binding of [(3)H]NAmide and IVM binds to GluCls. Finally, the binding of [(3)H]IVM is inhibited by NA. The B(max) of [(3)H]IVM is twice that of [(3)H]NAmide, and about half of the [(3)H]IVM binding sites are inhibited by NA with high affinity (K(I) = 25 pM). In contrast, [(3)H]IVM binding to Caenorhabditis elegans membranes is not inhibited by NA at 100 nM, and there are no high affinity binding sites for NA on these membranes. Thus, half of the Drosophila IVM receptors and all of the NA receptors are associated with GluCl. NA distinguishes between nematode and insect GluCls and identifies subpopulations of IVM binding sites. PMID- 10820029 TI - A conserved negatively charged amino acid modulates function in human nonmuscle myosin IIA. AB - A myosin surface loop (amino acids 391-404) is postulated to be an important actin binding site. In human beta-cardiac myosin, mutation of arginine-403 to a glutamine or a tryptophan causes hypertrophic cardiomyopathy. There is a phosphorylatable serine or threonine residue present on this loop in some lower eukaryotic myosin class I and myosin class VI molecules. Phosphorylation of the myosin I molecules at this site regulates their enzymatic activity. In almost all other myosins, the homologous residue is either a glutamine or an aspartate, suggesting that a negative charge at this location is important for activity. To study the function of this loop, we have used site-directed mutagenesis and baculovirus expression of a heavy meromyosin- (HMM-) like fragment of human nonmuscle myosin IIA. An R393Q mutation (equivalent to the R403Q mutation in human beta-cardiac muscle myosin) has essentially no effect on the actin activated MgATPase or in vitro motility of the expressed HMM-like fragment. Three mutations, D399K, D399A, and a deletion mutation that removes residues 393-402, all decrease both the V(max) of the actin-activated MgATPase by 8-10-fold and the rate of in vitro motility by a factor of 2-3. The K(ATPase) of the actin activated MgATPase activity and the affinity constant for binding of HMM to actin in the presence of ADP are affected by less than a factor of 2. These data support an important role for the negative charge at this location but show that it is not critical to enzymatic activity. PMID- 10820030 TI - Stereochemical outcome and kinetic effects of Rp- and Sp-phosphorothioate substitutions at the cleavage site of vaccinia type I DNA topoisomerase. AB - To probe the mechanism of the reversible DNA phosphodiester bond cleavage and religation mechanism of the type I topoisomerase from vaccinia virus, we have synthesized DNA substrates carrying a single nonbridging Rp- or Sp phosphorothioate (Ps) modification at the scissile phosphodiester (Pd) bond. Analysis of the stereochemical outcome of the net cleavage and rejoining reaction established that the reaction proceeds with retention of configuration, as expected for a double-displacement mechanism. Single-turnover kinetic studies on irreversible strand cleavage using 18/24 mer suicide substrates showed thio effects (k(Pd)/k(Ps)) of 340- and 30-fold for the Rp-Ps and Sp-Ps stereoisomers, respectively, but approximately 10-fold smaller thio effects for the reverse single-turnover religation reaction (Rp-Ps = 30 and Sp-Ps = 3). As compared to the smaller suicide cleavage substrates, approach-to-equilibrium cleavage studies using 32/32 mer substrates showed 7-9-fold smaller thio effects on cleavage, similar effects on religation, and the same ratio of the Rp to Sp thio effect as the suicide cleavage reaction ( approximately 10). In general, thio effects of 2.4-7.2-fold on the cleavage equilibrium are observed for the wild-type and H265A enzymes, suggesting differences in the interactions of the enzyme with the nonbridging sulfur in the noncovalent and covalent complexes. Studies of the cleavage, religation, and approach-to-equilibrium reactions catalyzed by the H265A active site mutant revealed a stereoselective, 11-fold decrease in the Rp thio effect on cleavage and religation as compared to the wild-type enzyme. This result suggests that His-265 interacts with the nonbridging pro-Rp oxygen in the transition state for cleavage and religation, consistent with the arrangement of this conserved residue in the crystal structure of the human topoisomerase-DNA complex. In general, the greatest effect of thio substitution and the H265A mutation is to destabilize the transition state, with smaller effects on substrate binding. The interaction of His-265 with the pro-Rp nonbridging oxygen is inconsistent with the proposal that this conserved residue acts as a general acid in the strand cleavage reaction. PMID- 10820031 TI - Regulation of rho-dependent transcription termination by NusG is specific to the Escherichia coli elongation complex. AB - To terminate transcription in E. coli, Rho protein binds an RNA loading site on the nascent transcript, translocates 5'--> 3' along the RNA in an ATP-driven process, and, upon reaching the transcription elongation complex, brings about RNA release. Thus, the Rho-dependent termination process can be viewed, in part, as a kinetic competition between the rate of transcript elongation by RNA polymerase (RNAP) and the rate of Rho translocation along the nascent transcript. In the context of this model, NusG, which is an essential E. coli protein, regulates Rho-dependent termination in an apparently paradoxical way, increasing the rate of transcription elongation of E. coli RNAP in the absence of Rho while also shifting the sites of Rho-dependent termination upstream on the template. Here we investigate the regulation of Rho-dependent termination by NusG. Analytical ultracentrifugation was used to establish the existence of a stable complex of NusG and Rho and to demonstrate a stoichiometry of one NusG monomer per Rho hexamer. Surface plasmon resonance was used to examine the kinetics of the formation and dissociation of the NusG-Rho complex, yielding an association rate constant (k(on)) of 2.8 (+/-0.8) x 10(5) M(-)(1) s(-)(1), a dissociation rate constant (k(off)) of 3.9 (+/-0.7) x 10(-)(3) s(-)(1), and a calculated equilibrium (dissociation) constant (K(d)) of 1.5 (+/-0.3) x 10(-)(8) M. The apparent stability of the NusG-Rho complex is insensitive to changes in salt (potassium acetate) concentration between 0.05 and 0.15 M. The translocation and transcription termination activities of Rho at saturating NusG concentrations were, however, both sensitive to salt concentration over this range, suggesting that these activities do not directly reflect the stability of the NusG-Rho complex. Rho-dependent termination could be demonstrated for transcription complexes in which E. coli RNAP had been substituted by either bacteriophage SP6 or T7 RNAP. NusG, however, was not active in transcription termination assays with either of these phage RNAPs. Thus, we conclude that NusG modulates Rho dependent termination by interacting specifically with the RNAP of the E. coli elongation complex to render the complex more susceptible to the termination activity of Rho. PMID- 10820033 TI - Interaction of cisplatin and DNA-targeted 9-aminoacridine platinum complexes with DNA. AB - Interaction of acridine- and 9-aminoacridinecarboxamide platinum complexes with DNA was investigated with respect to their DNA sequence specificity and kinetics of binding. The DNA sequence specificity of the compounds was quantitatively analyzed using a polymerase stop assay with the plasmid pUC19. The 9 aminoacridinecarboxamide platinum complexes exhibited a different sequence specificity to that of cisplatin, shifted away from runs of consecutive guanines (the main binding site for cisplatin). This alteration was dependent on chain length. Shorter chain length compounds (n = 2, 3) showed a greater difference in sequence specificity, while longer chain length compounds (n = 4, 5) more closely resembled cisplatin. An acridinecarboxamide platinum complex showed a similar sequence specificity to cisplatin, revealing that the major change of sequence specificity was due to the presence of the 9-amino substituent. A linear amplification system was used to investigate the time course of the reaction. The presence of an intercalating group (acridinecarboxamide or 9 aminoacridinecarboxamide) greatly increased the rate of reaction with DNA; this is proposed to be due to a different reaction mechanism with DNA (direct displacement by the N-7 of guanine). PMID- 10820032 TI - Novel substrates of Escherichia coli nth protein and its kinetics for excision of modified bases from DNA damaged by free radicals. AB - Escherichia coli Nth protein (endonuclease III) is a DNA glycosylase with a broad substrate specificity for pyrimidine derivatives. We discovered novel substrates of E. coli Nth protein using gas chromatography/isotope-dilution mass spectrometry and DNA samples, which were damaged by gamma-irradiation or by H(2)O(2)/Fe(III)-EDTA/ascorbic acid. These were 4, 6-diamino-5 formamidopyrimidine, 5,6-dihydroxyuracil, and 5, 6-dihydroxycytosine. The first compound was recognized for the first time as a purine-derived substrate of the enzyme. We also investigated kinetics of excision of a multitude of modified bases from three damaged DNA substrates. Excision of modified bases was determined as a function of enzyme concentration, incubation time, and substrate concentration. Excision followed Michaelis-Menten kinetics. Kinetic parameters were determined for the following modified bases: 4,6-diamino-5 formamidopyrimidine, cis- and trans-thymine glycols, 5-hydroxycytosine, cis- and trans-uracil glycols, 5-hydroxyuracil, 5-hydroxy-5-methylhydantoin, alloxan, 5, 6 dihydroxycytosine, 5,6-dihydroxyuracil, 5-hydroxy-6-hydrothymine, and 5-hydroxy-6 hydrouracil. The results show that three newly discovered substrates were excised by the enzyme with a preference similar to excision of its known major substrates such as thymine glycol and 5-hydroxycytosine. Excision kinetics significantly depended on the nature of the damaged DNA substrates in agreement with previous results on other DNA glycosylases. Specificity constants (k(cat)/K(M)) of E. coli Nth protein were compared to those of its previously investigated functional homologues such as human and Schizosaccharomyces pombe Nth proteins and Saccharomyces cerevisiae Ntg1 and Ntg2 proteins. This comparison shows that significant differences exist with respect to substrate specificity and kinetic parameters despite extensive structural conservation among the Nth homologues. PMID- 10820034 TI - Tyrosine residues serve as proton donor in the catalytic mechanism of epoxide hydrolase from Agrobacterium radiobacter. AB - Epoxide hydrolase from Agrobacterium radiobacter catalyzes the hydrolysis of epoxides to their diols via an alkyl-enzyme intermediate. The recently solved X ray structure of the enzyme shows that two tyrosine residues (Tyr152 and Tyr215) are positioned close to the nucleophile Asp107 in such a way that they can serve as proton donor in the alkylation reaction step. The role of these tyrosines, which are conserved in other epoxide hydrolases, was studied by site-directed mutagenesis. Mutation of Tyr215 to Phe and Ala and mutation of Tyr152 to Phe resulted in mutant enzymes of which the k(cat) values were only 2-4-fold lower than for wild-type enzyme, whereas the K(m) values for the (R)-enantiomers of styrene oxide and p-nitrostyrene oxide were 3 orders of magnitude higher than the K(m) values of wild-type enzyme, showing that the alkylation half-reaction is severely affected by the mutations. Pre-steady-state analysis of the conversion of (R)-styrene oxide by the Y215F and Y215A mutants showed that the 1000-fold elevated K(m) values were mainly caused by a 15-40-fold increase in K(S) and a 20 fold reduction in the rate of alkylation. The rates of hydrolysis of the alkyl enzyme intermediates were not significantly affected by the mutations. The double mutant Y152F+Y215F showed only a low residual activity for (R)-styrene oxide, with a k(cat)/K(m) value that was 6 orders of magnitude lower than with wild-type enzyme and 3 orders of magnitude lower than with the single tyrosine mutants. This indicates that the effects of the mutations were cumulative. The side chain of Gln134 is positioned in the active site of the X-ray structure of epoxide hydrolase. Mutation of Gln134 to Ala resulted in an active enzyme with slightly altered steady-state kinetic parameters compared to wild-type enzyme, indicating that Gln134 is not essential for catalysis and that the side chain of Gln134 mimics bound substrate. Based upon this observation, the inhibitory potential of various unsubstituted amides was tested, resulting in the identification of phenylacetamide as a competitive inhibitor with an inhibition constant of 30 microM. PMID- 10820036 TI - Critical role of tyrosine 277 in the ligand-binding and transactivating properties of retinoic acid receptor alpha PMID- 10820035 TI - Solution structure by NMR and molecular dynamics of a duplex containing a guanine opposite a N-(2-deoxy-beta-D-erythro-pentofuranosyl)formamide lesion. AB - One- and two-dimensional NMR spectroscopy has been used combined with molecular dynamics to determine the fine structure of the DNA duplex 5' d(AGGAGCCACG).d(CGTGGFTCCT) where F is the N-(2-deoxy-beta-D-erythro pentofuranosyl)formamide residue which is a ring fragmentation product of thymine. The formamide deoxyribose exists as two isomers with respect to the orientation about the peptide bond. The two isomers (trans and cis) are observed in a ratio 3:2 in solution. For both species, the oligonucleotide adopts a globally B form structure although conformational changes are observed around the mismatch site. The formamide residue, whatever the isomer, is intrahelical and can pair with the guanine on the opposite strand with one hydrogen bond. For the cis isomer, the residue adopts a syn orientation and is able to form a second hydrogen bond with the guanine on the 5' side on the same strand. Off-resonance ROESY experiments have been used to investigate the chemical exchange observed at low temperature of the duplex. Conformational exchange has only been found for the oligonucleotide with the formamide residue in the trans conformation. PMID- 10820037 TI - Lipid oxidation in a chicken muscle model system: oxidative response of lipid classes to iron ascorbate or methemoglobin catalysis. AB - Catalysis by iron ascorbate and activated methemoglobin generated different oxidative responses in chicken muscle model systems. In iron ascorbate systems, large increases in hydroperoxides and thiobarbituric acid-reactive substances (TBARS) occurred during the initial stage of incubation. Thereafter, iron ascorbate catalysis led to a slow increase in the oxidation of triacylglcyerol (TG) and sarcoplasmic reticulum (SR) membrane lipids. By the end of incubation, 24, 36, and 32% of the initial content of n-3 fatty acids in free fatty acids, TG, and SR single-lipid model systems catalyzed by iron ascorbate had been lost. Reduced losses of n-3 fatty acids were observed in the SR and TG fractions (0 and 24%, respectively) when iron ascorbate model systems contained all three lipid fractions (mix). Hydroperoxides and TBARS in model systems catalyzed by activated methemoglobin were characterized by a lag phase during most of the incubation. Consistent with their role as antioxidants, losses of alpha-tocopherol (42-49%), gamma-tocopherol (36-42%), and protein sulfhydryls (41-52%) were observed in model systems catalyzed by activated methemoglobin. SR and mix model systems were 30-50% slower to oxidize than TG model systems when activated methemoglobin served as the catalytic agent. PMID- 10820038 TI - Influence of dietary fat and vitamin E supplementation on free radical production and on lipid and protein oxidation in turkey muscle extracts. AB - The objectives of this study were to investigate the effects of dietary fat (6% soy oil or rapeseed oil or tallow) and alpha-tocopheryl acetate supplementation at two levels (30 or 200 ppm) on radical production, measured by ESR spectroscopy, and on lipid and protein oxidation in turkey muscle extracts oxidized by an enzymic system (NADPH, ADP, FeSO(4)/cytochrome P450 reductase). Two muscles were tested: pectoralis major (glycolytic) and sartorius (oxidative) muscles. Radical production measured by ESR was higher in pectoralis major muscle than in sartorius muscle, whereas lipid and protein oxidation was more important in sartorius muscle, showing the importance of the pro-/antioxidant ratio in oxidative processes in muscular cells and of the measurement methodology to appreciate the free radical production. Dietary fat had no effect on the level of ESR signals, whereas feeding of animals with soy oil induced higher oxidation of lipids. Protein oxidation was less sensitive to the nature of the dietary fat than lipid oxidation. Vitamin E supplementation significantly decreased radical production, as measured by ESR spectroscopy. Vitamin E also decreased lipid and protein oxidation, but the effect of vitamin E on protein oxidation was less pronounced than on lipid oxidation. PMID- 10820039 TI - Effect of high-pressure treatment on the texture of cherry tomato. AB - The effect of high-pressure treatment (200-600 MPa for 20 min) on the texture of cherry tomatoes and on the key softening enzymes (pectinmethylesterase and polygalacturonase) was investigated. When subjected to high-pressure treatment whole cherry tomatoes showed increasing textural damage with increasing pressures up to 400 MPa. However, treatment at pressures above 400 MPa (500-600 MPa) led to less apparent damage than treatment at 300 and 400 MPa; the tomatoes appearing more like the untreated samples. These visual changes were reflected in the texture (firmness) and amount of cell rupture in the tomatoes, with the least firmness and the most cell rupture being seen after treatment at 400 MPa. Light and scanning electron microscopy supported these observations. Although a sample of purified commercial pectinmethylesterase was partially inactivated at pressures above 200 MPa, irrespective of pH (4-9), in the whole cherry tomatoes no significant inactivation was seen even after treatment at 600 MPa, presumably because other components in the tomato offered protection or the isoenzymes were different. Polygalacturonase was more susceptible to pressure, being almost totally inactivated after treatment at 500 MPa. It is concluded that the textural changes in tomato induced by pressure involve at least two related phenomena. Initially, damage is caused by the greater compressibilty of the gaseous phase (air) compared to liquid-solid components, giving rise to a compact structure which, on pressure release, is damaged as the air rapidly expands, leading to increases in membrane permeability. This permits egress of water, and the damage also enables enzymatic action to increase, causing further cell damage and softening. The major enzyme involved in the further softening is polygalacturonase, which is inactivated at 500 MPa and above, and not pectinmethylesterase, which in the whole fruit, is barotolerant. PMID- 10820040 TI - Antioxidant activity of tomato products as studied by model reactions using xanthine oxidase, myeloperoxidase, and copper-induced lipid peroxidation. AB - The antioxidant content and activity of commercial tomato products differing in variety and processing were studied. Two procedures for extracting hydrophilic and lipophilic antioxidants, namely, two-step 0.1 M phosphate buffer (pH 3.0 and 7.4) extraction and tetrahydrofuran extraction followed by petroleum ether fractionation, were developed. Carotenoids (lycopene, beta-carotene, and lutein) and ascorbic acid were analyzed by HPLC with spectrophotometric and electrochemical detectors, respectively. Total phenolics were determined by using the Folin-Ciocalteu reagent. The antioxidant activity was studied by the following three model systems: (a) the xanthine oxidase (XOD)/xanthine system, which generates superoxide radical and hydrogen peroxide; (b) the myeloperoxidase (MPO)/NaCl/H(2)O(2) system, which produces hypochloric acid; and (c) the linoleic acid/CuSO(4) system, which promotes lipid peroxidation. Results showed that the hydrophilic and lipophilic fractions of all tomato products were able to affect model reactions, whatever reactive oxygen species and catalysts were used to drive oxidation. In the XOD/xanthine system both the hydrophilic and lipophilic fractions displayed an inhibitory activity. The hydrophilic fractions were more effective (I(50) ranging from 680 to 3200 microg, dry weight) than the lipophilic fractions (I(50) ranging from 4000 to 7750 microg, dry weight). In the MPO/NaCl/H(2)O(2) system the hydrophilic fractions inhibited oxidation (I(50) ranging from 2300 to 2900 microg, dry weight), whereas the lipophilic fractions had a lower inhibitory effect at the same concentration. Conversely, in the copper-catalyzed lipid peroxidation only the lipophilic fractions were effective (I(50) ranging from 1030 to 2100 microg, dry weight), whereas the hydrophilic fractions had a pro-oxidant effect in the same concentration range. The extent of inhibition varied according to the tomato sample in the superoxide and hydrogen peroxide generating system and in lipid peroxidation, but was substantially the same in the HClO generating system. Fresh tomato varieties differed considerably in the antioxidant activities of their hydrophilic and lipophilic fractions. Processed tomatoes showed a significantly lower antioxidant activity than fresh tomatoes in their hydrophilic fractions but had a high antioxidant activity in their lipophilic fractions. Because the oxidative reactions produced by the above mentioned model systems are also involved in the pathogenesis of several chronic diseases, the antioxidant activity of tomato fractions might be related to their in vivo activity. Hence, these measurements may be used for optimizing tomato technologies. PMID- 10820041 TI - In vitro antioxidant and ex vivo protective activities of green and roasted coffee. AB - The antioxidant properties of green and roasted coffee, in relation to species (Coffea arabica and Coffea robusta) and degree of roasting (light, medium, dark), were investigated. These properties were evaluated by determining the reducing substances (RS) of coffee and its antioxidant activity (AA) in vitro (model system beta-carotene-linoleic acid) and ex vivo as protective activity (PA) against rat liver cell microsome lipid peroxidation measured as TBA-reacting substances. RS of C. robustasamples were found to be significantly higher when compared to those of C. arabica samples (p < 0.001). AA for green coffee samples were slightly higher than for the corresponding roasted samples while PA was significantly lower in green coffee compared to that of all roasted samples (p < 0.001). Extraction with three different organic solvents (ethyl acetate, ethyl ether, and dichloromethane) showed that the most protective compounds are extracted from acidified dark roasted coffee solutions with ethyl acetate. The analysis of acidic extract by gel filtration chromatography (GFC) gave five fractions. Higher molecular mass fractions were found to possess antioxidant activity while the lower molecular mass fractions showed protective activity. The small amounts of these acidic, low molecular mass protective fractions isolated indicate that they contain very strong protective agents. PMID- 10820042 TI - Antioxidant properties of crocin from Gardenia jasminoides Ellis and study of the reactions of crocin with linoleic acid and crocin with oxygen. AB - Crocin-a water soluble carotenoid-is found in the fruits of gardenia (Gardenia jasminoides Ellis) and in the stigmas of saffron (Crocus sativus Linne). For crocin purification, gardenia fruits are extracted with 50% acetone, followed by ether washing, ion exchange, and separation by preparative HPLC. Purified crocin with purity of >99.6% has an antioxidative activity at concentrations up to 40 ppm. At 20 ppm the antioxidative activity of crocin is comparable to that of BHA. The antioxidant property of crocin as evaluated by the thiocyanate method was better than with the thiobarbituric acid method. The adduct between the linoleic acid radical and crocin was detected by LC-MS. When crocin reacted with oxygen in the presence of FeSO(4), intermediates such as monohydroperoxides and dihydroperoxides of crocin were formed and detected by LC-MS. PMID- 10820043 TI - In vivo antioxidative activity of propolis evaluated by the interaction with vitamins C and E and the level of lipid hydroperoxides in rats. AB - In vivo antioxidative activity of propolis was evaluated on the basis of ameliorative effects on the oxidative stress induced by vitamin E deficiency in rats. The control group was fed vitamin E-deficient diet, and the propolis group was fed vitamin E-deficient diet supplemented with 1% of propolis for 4 and 8 weeks. Comparisons were made in tissue concentrations of vitamin C, vitamin E, and lipid hydroperoxides between these groups. No significant difference was observed in tissue vitamin E concentration between these groups after both 4 and 8 weeks. After 4 weeks, the plasma vitamin C concentration of the propolis group was significantly higher than that of the control group. After 8 weeks, the tissue concentrations of vitamin C in the kidney, stomach, small intestine, and large intestine of the propolis group were significantly higher than those of the control group. These results suggest that some components of propolis are absorbed to circulate in the blood and behave as a hydrophilic antioxidant that saves vitamin C. The concentration of lipid hydroperoxides in the large intestine of the propolis group was significantly lower than that of the control group after 8 weeks. These results suggest that propolis exerts its antioxidative effect where it is assumed to accumulate, such as on the kidney, where it is excreted, and on the gastrointestinal tract, where propolis influences these tissues even from the outside of the cell. PMID- 10820044 TI - Studies on the antioxidant activity of Echinacea root extract. AB - Methanol extracts of freeze-dried Echinacea (E. angustifolia, E. pallida, and E. purpurea) roots were examined for free radical scavenging capacities and antioxidant activities. Root extracts of E. angustifolia, E. pallida, and E. purpurea were capable of scavenging hydroxyl radical. Similar scavenging activities for each variety were found for both 1,1-diphenyl-2-picrylhydrazyl radical and ABTS radical. Meanwhile, antioxidant activities of all three varieties of Echinacea were found to delay the formation of conjugated diene hydroperoxide induced by the thermal decomposition of 2, 2'-azobis(2 amidinopropane) dihydrochloride and extend the lag phase of peroxidation of soybean liposomes. Echinacea root extracts suppressed the oxidation of human low density lipoprotein, as evaluated by reduced agarose electrophoretic mobility following oxidative modification by Cu(2+). The mechanisms of antioxidant activity of extracts derived from Echinacea roots included free radical scavenging and transition metal chelating. PMID- 10820045 TI - Mechanisms of the antioxidant activity of a high molecular weight fraction of whey. AB - The antioxidant mechanisms of whey proteins in a Tween 20-stabilized salmon oil in-water emulsion were investigated. The antioxidant activity of the high molecular weight (HMW) fraction of whey from pasteurized milk was found to increase with concentration, as determined by its ability to inhibit TBARS and lipid peroxide formation. The ability of sulfhydryl-blocked whey to inhibit TBARS formation was reduced 60% compared to the HMW fraction alone at 7 days of storage. HMW fraction was able to scavenge peroxyl radicals, with scavenging decreasing approximately 20% when sulfhydryls were blocked. HMW fraction was able to chelate iron away from the surface of negatively charged BSA-stabilized emulsion droplets, indicating that the whey proteins were able to chelate iron. A better understanding of the mechanisms by which whey proteins inhibit lipid oxidation could increase the use of whey proteins as food antioxidants. PMID- 10820046 TI - Isolation and structure determination of new antioxidative ferulic acid glucoside esters from the rhizome of Alpinia speciosa, a Zingiberaceae plant used in Okinawan food culture. AB - An assay-guided isolation gave three antioxidants including two newly identified compounds from the rhizomes of Alpinia speciosa, which is used as an important plant in the food culture of the Okinawa area of Japan. Spectroscopic analysis of the two new compounds revealed them to be new glucoside esters of ferulic acid. The antioxidant activity of the esters was measured using two different methods. Both compounds showed greater activity than that of Trolox in the TLC method; however, one of the compounds showed weaker inhibitory activity than that of Trolox and epicatechin against AMVN-induced methyl linoleate oxidation. PMID- 10820047 TI - Changes in antioxidant effects and their relationship to phytonutrients in fruits of sea buckthorn (Hippophae rhamnoides L.) during maturation. AB - Different fractions of sea buckthorn fruits were investigated for antioxidant activity and its relationship to different phytonutrients. Capacity to scavenge radicals of the crude extract, like the phenolic and ascorbate extracts, decreased significantly with increased maturation. The changes were strongly correlated with the content of total phenolics and ascorbic acid. Antioxidant capacity of the lipophilic extract increased significantly and corresponded to the increase in total carotenoids. The phenolic fractions made a major contribution to the total antioxidant capacity due to the high content of total phenolics. The lipophilic fractions were most effective if the comparison was based on the ratio between antioxidant capacity and content of antioxidants. The crude extract of fruits showed the highest inhibitory effect in both 2,2 azobis(2,4-dimethylvaleronitrile) (AMVN) and ascorbate-iron induced lipid peroxidations. The aqueous and ascorbate-free extracts showed higher inhibition in the AMVN assay, but lower inhibition in ascorbate-iron induced peroxidation, than the lipophilic extract. PMID- 10820048 TI - Interfacial shear rheology of aged and heat-treated beta-lactoglobulin films: displacement by nonionic surfactant. AB - Interfacial shear rheology of adsorbed beta-lactoglobulin films (bulk protein concentration 10(-)(3) wt %) has been studied over the temperature range 20-90 degrees C using a two-dimensional Couette-type viscometer. Effects of the type of interface (air-water, triolein-water, and n-dodecane-water), the pH (2.0, 5.6, 6.0, 7.0, and 9.0), and the extent of the heat treatment have been assessed via measurements of changes in the apparent interfacial shear viscosity and elasticity before and after the addition of increasing amounts of nonionic surfactant Tween 20 (polyoxyethylene sorbitan monolaurate). The highest interfacial viscosities were obtained at the n-dodecane-water interface and the lowest at the air-water interface. Competitive displacement of protein from the interface by Tween 20 was easier at the air-water and n-dodecane-water interfaces as compared to the triolein-water interface. The surface shear viscosity was higher and the displacement by Tween 20 more difficult as the isoelectric point of the protein was approached, which is in agreement with the presence of a more strongly cross-linked protein network at the interface. The effect of heat treatment was dependent on the pH of the aqueous solution. No simple relationship between the surface rheological characteristics and the ease of displacement by Tween 20 could be inferred. PMID- 10820049 TI - Identification of flavonoid markers for the botanical origin of Eucalyptus honey. AB - European Eucalyptus honeys showed a common and characteristic HPLC profile in which the flavonoids myricetin (3,5,7,3',4', 5'-hexahydroxyflavone), tricetin (5,7,3',4',5'-pentahydroxyflavone), quercetin (3,5,7,3',4'-pentahydroxyflavone), luteolin (5,7,3', 4'-tetrahydroxyflavone), and kaempferol (3,5,7, 4' tetrahydroxyflavone) were identified. Their contents, and relative amounts, in the analyzed honey samples were quite constant and supported their floral origin. In addition, ellagic acid and the propolis-derived flavonoids pinobanksin, pinocembrin, and chrysin were detected in most samples. The contents of these nonfloral phenolics were much more variable as could be expected for their propolis origin. Myricetin, tricetin, and luteolin had not been identified as floral markers in any other honey sample previously analyzed in our laboratory (chestnut, citrus, rosemary, lavender, acacia, rapeseed, sunflower, heather, lime tree, etc.) or reported in the literature, suggesting that these could be useful markers. Only in some individual heather samples produced in Portugal has tricetin previously been detected in minor amounts. These samples, however, were contaminated with Eucalyptus as revealed by their pollen analysis and the lack of tricetin or their glycosides in heather floral nectar. It remains to be established if myricetin, tricetin, and luteolin originate from Eucalyptus floral nectar where the corresponding glycosides should be present. PMID- 10820050 TI - Bactericidal action of egg yolk phosvitin against Escherichia coliunder thermal stress. AB - Chicken egg yolk phosvitin showed a remarkable antibacterial effect against Escherichia coli under thermal stress at 50 degrees C. E. coli cells (10(6)/mL) completely disappeared in 1 mL of L-broth coexisting with 0.l mg/mL phosvitin when incubated at 50 degrees C for 20 min, whereas a considerable amount of cells (10(5)/mL) survived at the same thermal stress without phosvitin. Blocking of the chelating effect of phosvitin by the addition of Ca(2+) ion displayed a protective effect against the bactericidal activity at 50 degrees C. In addition, the antibacterial activity of phosvitin was dramatically reduced by treatment with alpha-chymotrypsin, although the chelating effect remained. The surface properties, such as interfacial tension and emulsifying properties of phosvitin, which are an index of the affinity with the outer membrane, were greatly reduced by the alpha-chymotrypsin digestion. This indicates that the alpha-chymotrypsin digested membrane-penetrating hydrophobic domains at the N- and C-terminal regions play an important role in antibacterial activity. These results suggest that a significant part of the bactericidal activity of phosvitin against E. coli resides in the synergistic effect of the high metal-chelating ability and the high surface activity under the influence of thermal stress. PMID- 10820051 TI - Objective measurement of red grapefruit juice color. AB - The color properties (CIE Lab, hue, and chroma) of 90 red grapefruit juices were studied by tristimulus reflectance measurement for two growing seasons. Juices were prepared from six red grapefruit cultivars including Ruby Red, Rio Red, Star Ruby, Ray Ruby, Flame, and Marsh Red grown in Florida. Very wide varietal and seasonal variations on juice color were observed, and most notably, CIE a values varied widely from -1.41 to 9.06 (CV = 82.6%). Lycopene is the major colored pigment in red cultivars with lesser amounts of beta-carotene as determined by HPLC on a carotenoid C(30) column with gradient elution using MeOH and MTBE. The highest correlation coefficients (r = 0.963) found between CIE a values and lycopene contents in juices from all cultivars combined suggest the possibility that the CIE a value would be a good indicator for pigmentation in red grapefruit juice. PMID- 10820052 TI - Origin of acetaldehyde during milk fermentation using (13)C-labeled precursors. AB - Acetaldehyde formation by Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus during fermentation of cow's milk was investigated using (13)C-labeled glucose, L-threonine, and pyruvate with a recent static-and trapped-headspace technique that does not require derivatization of acetaldehyde prior to gas chromatography-mass spectrometry. Over 90% and almost 100% of acetaldehyde originated from glucose during fermentation by L. delbrueckii subsp. bulgaricus and S. thermophilus, respectively, taking into account both singly and doubly labeled acetaldehyde. As both microorganisms showed threonine aldolase activity and formed labeled acetaldehyde from (13)C-labeled threonine during the fermentation of milk, this amino acid should also contribute to the acetaldehyde produced. PMID- 10820053 TI - Structure-activity relationships of (1'S)-1'-acetoxychavicol acetate, a major constituent of a southeast Asian condiment plant Languas galanga, on the inhibition of tumor-promoter-induced Epstein-Barr virus activation. AB - The structure-activity relationships of (1'S)-1'-acetoxychavicol acetate (ACA), a cancer chemopreventive agent of food origin, were investigated in an inhibitory test of tumor promoter teleocidin B-4-induced Epstein-Barr virus (EBV) activation in Raji cells. Through a test of 16 derivatives, the structural factors regulating activity were found to be as follows: (1) the absolute configuration at the 1'-position does not affect activity; (2) hydrogenation of the terminal methylene group abolishes activity; (3) both the phenolic and alcoholic hydroxyl groups are compulsorily acetylated, and it is necessary that the former is oriented only at the position para to the side chain; (4) an additional acetoxyl group is allowed to locate at the ortho or meta position; and (5) substitution of the hydrogen atom at the 1'-position by a methyl group reduces activity. Upon esterase blockade in Raji cells, (1'R,S)-ACA suppressed EBV activation, the extent of which was the same as tested in the control, suggesting that ACA bearing two acetoxyl groups is an intracellular structure prerequisite for activity exhibition. The present study suggests that nucleophilic attack to the 3'-position is important and involved in the interaction of ACA with an unidentified target molecule(s) participating in the process of EBV activation. PMID- 10820054 TI - Study of the compositional changes of mango during ripening by use of nuclear magnetic resonance spectroscopy. AB - Liquid-state NMR spectroscopy was used to follow the compositional changes in mango juice during ripening, whereas MAS and HR-MAS techniques enabled resolved (13)C and (1)H NMR spectra of mango pulps to be recorded. Spectral assignment enabled the identification of several organic acids, amino acids, and other minor components, and the compositional changes upon ripening were followed through the changes in the spectra. In pulps, sucrose was found to predominate over fructose and glucose at most ripening stages, and citric acid content decreased markedly after the initial ripening stages while alanine increased significantly. Other spectral changes reflect the complex biochemistry of mango ripening and enabled the role played by some compounds to be discussed. Some differences observed between the composition of juices and pulps are discussed. This work shows that NMR spectroscopy enables the direct characterization of intact mango pulps, thus allowing the noninvasive study of the overall biochemistry in the whole fruit. PMID- 10820055 TI - Purification and kinetic characterization of an anionic peroxidase from melon (Cucumis melo L.) cultivated under different salinity conditions. AB - The partial characterization of an anionic peroxidase in melon fruit is described. Four melon peroxidase (MPX) isoenzymes were detected in crude extracts after isoelectric focusing. The major MPX isoenzyme (pI = 3.7) was partially purified by including hydrophobic and anion-exchange chromatography in the purification scheme. The sample obtained was used to characterize MPX. This peroxidase did not show activity on ascorbic acid but oxidized guaiacol at a high rate, showing an optimum pH of 5.5 when acting on this last reducing substrate. Melon fruits grown under highly saline conditions showed slightly increased levels of this anionic isoenzyme. Kinetic studies using 2,2'-azinobis(3 ethylbenzothiazolinesulfonic acid) (ABTS) as reducing substrate showed that increased salinity in the growth medium did not modify the kinetic parameters of melon peroxidase on both hydrogen peroxide and reducing substrate. PMID- 10820056 TI - Raman spectroscopic study of oat globulin conformation. AB - Analysis of Raman spectra of oat globulin showed that extreme pH values caused an increase in the amide and C-H stretching band intensity, indicating changes in the secondary structures of the protein due to denaturation. Similar changes were observed when oat globulin was treated with chaotropic salts and several protein perturbants. Sodium dodecyl sulfate, beta-mercaptoethanol, and ethylene glycol also caused a shift in the amide III' band, suggesting a transition from beta sheet to a random coil conformation. Heating at temperatures near the denaturation temperature of oat globulin led to increases in the amide and C-H band intensity, indicating unfolding of the protein. The data indicate that FT Raman spectroscopy is suitable for studying the secondary structure of plant proteins such as oat globulin. PMID- 10820057 TI - Formation of new protein structures in heated mixtures of BSA and alpha lactalbumin. AB - The heat-induced protein-protein interactions of alpha-lactalbumin (alpha-La) and bovine serum albumin (BSA), dispersed in a pH 6.8, 10% whey protein concentrates (WPC) permeate, were followed using alkaline and sodium dodecyl sulfate (SDS) 1D and 2D polyacrylamide gel electrophoresis (PAGE) and size-exclusion high performance liquid chromatography (SE-HPLC). Heated (75 degrees C) 5% BSA solution contained large disulfide-bonded BSA aggregates, although some monomer BSA (SDS-monomeric BSA) could be dissociated from the aggregates by SDS. In contrast, similarly heated alpha-La solutions contained small quantities of several monomeric forms of alpha-La and dimeric alpha-La but no large aggregates. When 10% solutions of 1:1 (w/w) mixtures of alpha-La and BSA were heated, large disulfide-bonded aggregates and SDS-monomeric BSA and alpha-La were present. However, heated 2% mixtures contained more modified alpha-La monomers, alpha-La dimers, and alpha-La-trimers, fewer large disulfide-bonded aggregates, and less SDS-monomeric alpha-La or BSA. These results suggest that BSA forms disulfide bonded aggregates that contain available thiol groups that can catalyze the formation of differently structured alpha-La monomers, dimers, higher polymers, and adducts of alpha-La with BSA. PMID- 10820058 TI - Novel nitric oxide and superoxide generation inhibitors, persenone A and B, from avocado fruit. AB - One known, (2R)-(12Z,15Z)-2-hydroxy-4-oxoheneicosa-12,15-dien+ ++-1-yl acetate (1), and two novel compounds, persenone A (2) and B (3), have been isolated from avocado fruit (Persea americana P. Mill), as inhibitors of superoxide (O(2)(-)) and nitric oxide (NO) generation in cell culture systems. They showed marked inhibitory activities toward NO generation induced by lipopolysaccharide in combination with interferon-gamma in mouse macrophage RAW 264.7 cells. Their inhibitory potencies of NO generation (1, IC(50) = 3.6; 2, IC(50) = 1.2; and 3, IC(50) = 3.5 microM) were comparable to or higher than that of a natural NO generation inhibitor, docosahexaenoic acid (DHA; IC(50) = 4.3 microM). Furthermore, compounds 1-3 and DHA markedly suppressed tumor promoter 12-O tetradecanoylphorbol-13-acetate-induced O(2)(-) generation in differentiated human promyelocytic HL-60 cells (1, IC(50) = 33.7; 2, IC(50) = 1.4; 3, IC(50) = 1.8; and DHA, IC(50) = 10.3 microM). It is notable that they were found to be suppressors of both NO- and O(2)(-)-generating biochemical pathways but not to be radical scavengers. The results indicate that these compounds are unique antioxidants, preferentially suppressing radical generation, and thus may be promising as effective chemopreventive agent candidates in inflammation associated carcinogenesis. PMID- 10820059 TI - Ascorbic acid and ascorbic acid 6-palmitate induced oxidation in egg yolk dispersions. AB - The oxidation in aqueous dispersions of egg yolk powder and the influence of addition of the proposed antioxidants ascorbic acid and ascorbic acid 6-palmitate indicate that both ascorbic acid and ascorbic acid 6-palmitate propagated the oxidation of egg yolk powder dispersions. Ascorbic acid 6-palmitate was found to be more prooxidative than ascorbic acid. Moreover, it was found that addition of ascorbic acid or ascorbic acid 6-palmitate gave rise to an increase in the amount of free iron Fe(II) in the egg yolk dispersions. It is proposed that ascorbic acid and ascorbic acid 6-palmitate react with the phosvitin-Fe(III) complex found in egg yolk and release Fe(II), which subsequently propagates lipid oxidation. It appears that less oxidation occurs in egg yolk dispersions exposed to high concentrations of peroxy radicals with added ascorbic acid than egg yolk dispersions with added ascorbic acid without exposure to peroxy radicals. PMID- 10820060 TI - Kinetics of Fe(III) reduction by ascorbic acid in aqueous solutions. AB - The reaction of Fe(III) and ascorbic acid (AA) in food products and digestive tracts affects the efficiency and uptake of these two nutrients. We investigated the kinetics of Fe(III) reduction by AA at pH 5 and 6 in a model system at 25 degrees C. The results indicate that the reduction of Fe(III) by AA is of zero order with respect to AA. The reaction order with respect to Fe(III) cannot be represented by a simple kinetic model at pH 5 or 6. The major stage of the reduction (about 80%, stoichiometrically), however, could be represented by a general equation of -d[Fe(III)]/dt = k[Fe(III)],(1. 811) where k is a rate constant and [Fe(III)] is the total ferric concentration. The rate constant decreased 1 order of magnitude as pH increased from 5 to 6. Aging of Fe(III) solution slows its reduction rate at pH 6 but not at pH 5. PMID- 10820061 TI - Purification and partial characterization of three turnip (Brassicanapus L. var. esculenta D.C.) peroxidases. AB - Three turnip peroxidases (fractions C1, C2, and C3) were partially purified and characterized, to permit study of their feasibility for use in clinical and enzyme immunoassays. These fractions represented 20% of the initial activity, and fractions C1 and C2 were purified to homogeneity. The optimum pH was between 5.0 and 5.5, while optimum temperature ranged from 40 to 55 degrees C. The ABTS K(m) values for the two acidic fractions (C2 and C3) were 0.70 and 0.42 mM, respectively; about 5 times lower than that reported for the acidic commercial horseradish peroxidase (HRP). Fraction C3 had 4 times higher K(m) value than commercial cationic HRP. The molecular weights determined by SDS-PAGE ranged from 39.2 to 42.5 kDa. Activation energies for inactivation were 113 (C1), 130 (C2), and 172 kJ/mol (C3) which are higher or comparable to other peroxidase isoenzymes reported. Fractions C1 and C3 represent an alternative source of peroxidase because of their higher purification yield and specific activity, when compared to fraction C2. PMID- 10820062 TI - Physicochemical properties and susceptibility to proteolytic digestion of myoglobin-phenol derivatives. AB - This paper deals with the interactions of chlorogenic, caffeic, and quinic acids and p-quinone with myoglobin. The myoglobin derivatives formed have been characterized in terms of physicochemical properties and susceptibility to proteolysis. The results show that the free amino group and tryptophan contents of the myoglobin-phenol derivatives decrease with the increasing extent to which the protein becomes derivatized. Furthermore, the solubility of myoglobin-phenol derivatives decreases in the pH range 3.5-6.5 as compared to solubility of the native protein. The reaction also influences the hydrophilic-hydrophobic character of the protein. The isoelectric point of the derivatized myoglobin is shifted to a lower pH value, and formation of high molecular fractions is also documented. This paper also demonstrates the influence of the protein derivatization with plant phenols on susceptibility to digestion by trypsin, alpha-chymotrypsin, and pepsin, determined in vitro. The enzymatic digestion of the derivatized proteins is adversely affected. PMID- 10820063 TI - Anthocyanin-based natural colorants: a new source of antiradical activity for foodstuff. AB - The antiradical capacity (radical scavenger capacity, RSC) of anthocyanin-based fruit extracts prepared in the laboratory (black chokeberry, black-thorn, and strawberry) was studied by using the 2, 2-diphenyl-1-picrylhydrazyl radical (DPPH(*)). To determine their RSC, the second-order rate constant (k(2)) for the oxidation of these extracts by DPPH(*) was calculated. The value of k(2) was compared to that used in the food industry as natural (alpha-tocopherol) or synthetic (butylated hydroxytoluene (BHT) and butylated hydroxyanisole (BHA)) antioxidants, as well as for a commercial elderberry concentrate and a synthetic colorant (Ponceau 4R). The k(2) values ((mg/mL)(-)(1) s(-)(1)), in methanol at 25 degrees C, were 1.87, 0.7, 0.42, 0.2, 0.05, 0.03, and 0.008 for alpha-tocopherol, black chokeberry, BHA, black-thorn, BHT, strawberry, and elderberry, respectively. Ponceau 4R lacked RSC. Therefore, these natural colorants proved to be a combined source of color and RSC for food material. PMID- 10820064 TI - Influence of sucrose on the thermal denaturation, gelation, and emulsion stabilization of whey proteins. AB - The influence of sucrose (0-40 wt %) on the thermal denaturation and functionality of whey protein isolate (WPI) solutions has been studied. The effect of sucrose on the heat denaturation of 0.2 wt % WPI solutions (pH 7.0) was measured using differential scanning calorimetry. Sucrose increased the temperature at which protein denaturation occurred, for example, by 6-8 degrees C for 40 wt % sucrose. The dynamic shear rheology of 10 wt % WPI solutions (pH 7.0, 100 mM NaCl) was monitored as they were heated from 30 to 90 degrees C and then cooled to 30 degrees C. Sucrose increased the gelation temperature and the final rigidity of the cooled gels. The degree of flocculation in 10 wt % oil-in-water emulsions stabilized by 1 wt % WPI (pH 7.0, 100 mM NaCl) was measured using a light scattering technique after they were heated at fixed temperatures from 30 to 90 degrees C for 15 min and then cooled to 30 degrees C. Sucrose increased the temperature at which maximum flocculation was observed and increased the extent of droplet flocculation. These results are interpreted in terms of the influence of sucrose on the thermal unfolding and aggregation of protein molecules. PMID- 10820065 TI - Influence of calcium, magnesium, or potassium ions on the formation and stability of emulsions prepared using highly hydrolyzed whey proteins. AB - Oil-in-water emulsions (4 wt % soy oil) containing 4 wt % whey protein hydrolysate (WPH) (27% degree of hydrolysis) and different levels of calcium, magnesium, or potassium chloride were prepared in a two-stage homogenizer. Other emulsions containing 4 wt % WPH but including 0.35 wt % hydroxylated lecithin and different levels of the above minerals were similarly prepared. The formation and stability of these emulsions were determined by measuring oil droplet size distributions using laser light scattering and by confocal scanning laser microscopy and a gravity creaming test. Both lecithin-free and lecithin containing emulsions showed no change in droplet size distributions with increasing concentration of potassium in the range 0-37.5 mM. In contrast, the diameter of emulsion droplets increased with increasing calcium or magnesium concentration >12.5 mM. Emulsions containing hydroxylated lecithin were more sensitive to the addition of calcium or magnesium than the lecithin-free emulsions. Storage of emulsions at 20 degrees C for 24 h further increased the diameter of droplets and resulted in extensive creaming in emulsions containing >25 mM calcium or magnesium. It appears that both flocculation and coalescence processes were involved in the destabilization of emulsions induced by the addition of divalent cations. PMID- 10820066 TI - Changes in tocopherol and plastochromanol-8 contents in seeds and oil of oilseed rape (Brassica napus L.) during storage as influenced by temperature and air oxygen. AB - The changes in tocopherol and plastochromanol-8 contents in seeds and oil of oilseed rape (Brassica napus L.) were studied during a storage period of 24 weeks at different incubation temperatures and exposure to air oxygen (open and closed flasks). In the extracted oil, total tocopherol content remained unaltered at 5 and 20 degrees C throughout the 24 weeks of storage. At 40 degrees C, a beginning degradation was observed already after 4 weeks in both open and closed flasks; the alpha-tocopherol content was affected most, followed by gamma-tocopherol and plastochromanol-8. After 16 weeks at 40 degrees C, the total tocopherol content in the oil was reduced by more than 90%. In intact seeds, no tocopherol degradation was observed; only the seeds incubated at 40 degrees C and in open flasks showed slightly lower tocopherol contents. However, the analysis of the tocopherol composition in the stored seeds showed a decrease in the alpha tocopherol content and an increase in the gamma-tocopherol content, which resulted in a decreasing alpha-/gamma-tocopherol ratio. This trend was most apparent at 40 degrees C and after 24 weeks of storage. A reduction of plastochromanol-8 occurred only at 40 degrees C and was more pronounced in open flasks. At 40 degrees C and in closed flasks a gradual increase in the content of alpha-tocotrienol was observed, a compound normally not accumulated in rapeseed. PMID- 10820067 TI - Characterization of commercial and experimental sodium caseinates by multiangle laser light scattering and size-exclusion chromatography. AB - A range of sodium caseinate samples were characterized by a multiangle laser light scattering (MALLS) system or by the use of MALLS as an on-line detector with size-exclusion chromatography (SEC). Sodium caseinate solutions, analyzed using a MALLS system alone, gave weight-average molar mass (M(w)) values in the range 1200-4700 kDa and z-average root-mean-square radius (R(g)) values ranged from approximately 50 to 120 nm. When these solutions were ultracentrifuged at 90000g for 1 h, a cloudy top layer was formed; the subnatant was carefully removed and analyzed by SEC-MALLS. The M(w) values were found to be in the range approximately 30-575 kDa, and R(g) values ranged from approximately 22 to 49 nm. During SEC, the MALLS system detected some very large-sized material that eluted close to the void volume; this material was hardly detected by the concentration detectors, i.e., ultra-violet (UV) and differential refractive index (DRI). The intensity of the light scattering (LS) signal from this very large sized material was greatly reduced in the subnatant. SEC of sodium caseinate samples revealed two main peaks with M(w) of approximately 420-750 kDa and 39-69 kDa, respectively. The R(g) values were very large for a protein molecule, and initial calculations suggested that the shape of caseinate molecules was likely to be highly elongated. PMID- 10820068 TI - Xanthophyll esterification accompanying carotenoid overaccumulation in chromoplast of Capsicum annuum ripening fruits is a constitutive process and useful for ripeness index. AB - Changes in xanthophyll esterification degree during pepper fruit ripening have been studied in five cultivars (Numex, Mana, Belrubi, Delfin, and Negral). Esterification of xanthophylls with fatty acids is seen to be a process that is contemporary with and directly linked to the transformation of chloroplast (present in the green fruit) into chromoplast (present in the red fruit). Changes in the fractions of free and partially and totally esterified carotenoids are similar between varieties, reflecting the constitutive nature of esterification as part of the ripening process and being controlled by it. From the first stages of ripening, the fraction of totally esterified pigments (zeaxanthin diester, beta-cryptoxanthin diester, capsanthin diester, and capsorubin diester) makes up almost 50% of the total carotenoid content. The proportion of the partially esterified pigment fraction (zeaxanthin monoester, capsanthin monoester, and capsorubin monoester) in the total carotenoid content increases, with a gradual decrease in the fraction of free pigments (beta-cryptoxanthin, beta-carotene, zeaxanthin, capsanthin, and capsorubin). In the fully ripe stage, a balance is reached between the three esterification fractions (free, partially esterified, and totally esterified), with mean values of 24.17 +/- 4.06, 31.48 +/- 4. 61, and 44.36 +/- 5.05, respectively, which seems to be largely independent of variety. This suggests a marked control of the carotenoid composition of the totally developed chromoplast, indicating its use as an index of ripeness. The inclusion in the present study of a variety (Negral) that retains chlorophylls when ripening, and which shows the same esterification behavior, supports the idea that carotenogenesis is normal and independent of chlorophyll catabolism. PMID- 10820069 TI - Oleuropein site selective hydrolysis by technomimetic nuclear magnetic resonance experiments. AB - Technomimetic NMR experiments were performed in accordance with the lye treatment adopted during table olive industrial procedures for the debittering process causing oleuropein degradation. The site selective hydrolysis of the two ester groups, characterizing the biophenolic secoiridoid molecule, was shown to be dependent on the different reactivities of these functionalities. The process is controlled by the experimental conditions exerted on the olive pulp and determined by the buffering capacity of the olive mesocarp and by the epicarp molecular components influencing the reactant penetration into the fruit pulp. The overall hydrolytic process of oleuropein, the bitter principle of olives, using the technomimetic experimental mode, gave rise to its catabolic derivatives, hydroxytyrosol, 11-methyloleoside, and the monoterpene glucoside, technomimetically produced, isolated, and structurally characterized by (1)H, (13)C, and COSY spectroscopy as the oleoside. PMID- 10820070 TI - Flavor enhancement of reduced fat cheddar cheese using an integrated culturing system. AB - Mild cheese flavor in reduced fat Cheddar cheese was enhanced by using an integrated starter culture system. Three cultures, Lactococcus lactis subsp. cremoris SK11, L. lactis subsp. lactis biovar. diacetylactis JVI, and Lactobacillus casei 7A, were carefully selected to obtain a nonbitter, mildly acid, buttery flavored cheese. Cheeses were produced from all possible combinations of these cultures with the constraint that L. lactis subsp. cremoris SK11 was used as the primary acid-producing culture. Cheeses made with SK11 were compared to cheeses produced using an L. lactis subsp. cremoris commercial starter culture. Cheeses were ripened for 150 days and periodically sampled for chemical, microbiological, and sensory analysis. Cheeses produced with L. lactis subsp. cremorisSK11 had substantially lower bitterness intensity than the cheeses produced with commercial starter culture. L. lactis subsp. lactis biovar. diacetylactis JVI significantly increased diacetylacetoin and acetate concentrations. Sensory results indicate that these cheeses had increased buttery (diacetyl) flavor. PMID- 10820071 TI - Thaumatin-like proteins and chitinases, the haze-forming proteins of wine, accumulate during ripening of grape (Vitis vinifera) berries and drought stress does not affect the final levels per berry at maturity. AB - Thaumatin-like proteins and chitinases, which are pathogenesis-related (PR) proteins, were the major soluble protein components of grapes from five cultivars of Vitis vinifera. This dominance of PR proteins was apparent at berry softening (veraison) and then throughout berry development for the Muscat of Alexandria, Sultana, and Shiraz cultivars and in the berries of the Sauvignon Blanc and Pinot Noir cultivars examined at commercial maturity. The M(r) of the major thaumatin like protein from Muscat of Alexandria grapes was 21 272, and those of the three major chitinases from this cultivar, ChitB, ChitC, and ChitD, were 25 588, 25 410, and 25 457, respectively. The vines in the study were irrigated and showed no obvious signs of disease. Shiraz vines that had not been irrigated throughout the season were clearly water stressed, but had levels of PR proteins in the berry similar to vines that had been fully irrigated. It appears that the production of PR proteins that cause protein instability in wines by grapes may be little influenced by environmental conditions. PMID- 10820072 TI - Fermentation of plant cell wall derived polysaccharides and their corresponding oligosaccharides by intestinal bacteria. AB - New types of nondigestible oligosaccharides were produced from plant cell wall polysaccharides, and the fermentation of these oligosaccharides and their parental polysaccharides by relevant individual intestinal species of bacteria was studied. Oligosaccharides were produced from soy arabinogalactan, sugar beet arabinan, wheat flour arabinoxylan, polygalacturonan, and rhamnogalacturonan fraction from apple. All of the tested substrates were fermented to some extent by one or more of the individual species of bacteria tested. Bacteroides spp. are able to utilize plant cell wall derived oligosaccharides besides their reported activity toward plant polysaccharides. Bifidobacterium spp. are also able to utilize the rather complex plant cell wall derived oligosaccharides in addition to the bifidogenic fructooligosaccharides. Clostridium spp., Klebsiella spp., and Escherichia coli fermented some of the selected substrates in vitro. These studies do not allow prediction of the fermentation in vivo but give valuable information on the fermentative capability of the tested intestinal strains. PMID- 10820073 TI - Water extract of defatted rice bran suppresses visceral fat accumulation in rats. AB - Rice bran has been reported to inhibit pancreatic lipase activity in vitro. This action shows that administration of rice bran may result in a decrease in plasma triglyceride levels and suppress accumulation of fat in vivo. We administered water extract of defatted rice bran (WED-rice bran) to rats to determine its effects. Single administration of WED-rice bran at a dose of 1 g/kg body weight caused a decrease in plasma triglyceride levels in fat emulsion induced hypertriglyceridemic rats. Four week administration of WED-rice bran suppressed accumulation of visceral fat and body weight gain without influencing food consumption, liver function, and renal function. These results indicate that a reduction of plasma triglycerides and suppression of visceral fat accumulation may be induced by pancreatic lipase inhibition caused by administration of WED rice bran. PMID- 10820074 TI - MALDI-TOF MS analysis of food flavonol glycosides. AB - Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a new technique that is having a great impact on food analysis. This study is the first to demonstrate the use of MALDI-TOF MS to identify flavonol glycosides in food samples. 2',4',6'-Trihydroxyacetophenone was chosen as the best matrix because it worked for crude sample extracts and ionized flavonol glycosides in both positive and negative MALDI-TOF MS modes. In the positive mode, multiple ion forms were observed for flavonol glycosides, including [M + H](+), [M + Na](+), [M + K](+), and [M - H + Na + K](+), with further fragmentation through loss of glycosidic residues. The negative mode for all flavonol glycosides resulted in [M - H](-) ion formation without detectable fragmentation. The multiple ions in the positive mode gave more information on individual flavonol glycoside structures than the negative mode. Flavonol glycosides showed similar intensities or responses in the positive mode, while kaempferol glycosides exhibited much less response than quercetin glycosides in negative mode. PMID- 10820075 TI - Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry of polygalloyl polyflavan-3-ols in grape seed extract. AB - Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was employed to characterize the polygalloyl polyflavan-3-ols (PGPF) in grape seed extracts. Masses corresponding to a series of PGPF units inclusive of nonamers were observed in the positive-ion reflectron mode. Masses of PGPF inclusive of undecamers were observed in the positive-ion linear mode, providing the first known evidence of PGPF of this size. Soluble PGPF of grape seed extracts were precipitated by complexation with Yb(3+). The PGPF were then recovered by dissolving the precipitate in water and removing the Yb(3+) by a weakly acidic cation-exchange resin (Amberlite IRP-64). Comparisons of HPLC chromatograms of the crude grape seed extract prior to precipitation with Yb(3+) and after recovery of the PGPF indicated that 96% of the phenolic compounds were precipitated and 99% of the precipitated PGPF were recovered by cation-exchange resin. These results indicate that MALDI-TOF MS is able to determine the mass distribution of complex mixtures of oligomeric PGPF and that precipitation of PGPF by Yb(3+) is useful for isolation and quantification. PMID- 10820076 TI - Supercritical fluid extraction of atrazine and other triazine herbicides from fortified and incurred eggs. AB - Triazines are a class of important pre-emergent weed herbicides. Some members of this class of herbicides exhibit carcinogenic and immunotoxicity properties, which make their use controversial in areas where animal feed crops are grown. It is therefore important to determine if triazine residues are transported to animal food products in order to ascertain the extent of human exposure. Most of the current herbicide residue extraction methods are time-consuming and solvent intensive. Supercritical fluid extraction (SFE) using CO(2) has been used as a alternative for other residue extraction methods as a replacement for hazardous organic solvents. In this study, 10 triazines were extracted from eggs fortified at 100 ppb using unmodified supercritical CO(2) at a pressure of 10000 psi and a temperature of 50 degrees C with off-line collection on a solid phase extraction cartridge containing Florisil. Atrazine recovery averaged 90.4% with an RSD of 3.3%. The other triazines were recovered at mean levels >73%. In a separate feeding study, atrazine and two of its dealkyl metabolites were detected in the egg. The results indicate that SFE is a viable technique for isolating triazine residues from eggs, requiring only 8 mL of solvent for each analysis. PMID- 10820077 TI - Liquid chromatographic analysis of incurred amoxicillin residues in catfish muscle following oral administration of the drug. AB - Improper application of antibiotic chemicals to livestock and aquaculture species may lead to the occurrence of residues in food supplies. An appropriate depletion period is needed after the administration of drugs to animals for ensuring that residues in edible tissues are below established tolerance levels. This study was conducted to determine incurred amoxicillin residues in catfish muscle following oral administration. Dosed fish were harvested after four depletion periods, and muscle fillets were analyzed for amoxicillin residues using an HPLC method with precolumn derivatization and fluorescence detection. The residue levels in fish after a 6-h depletion ranged from 40 to 64 ng/g with one exception at 297 ng/g. Average residue levels decreased to 5.4 and 2. 8 ng/g after 24- and 48-h depletions, respectively. Residue levels after a 72-h depletion decreased to below the method's limit of quantitation (1.2 ng/g). An LC-MS/MS confirmatory method was developed. Confirmation of the presence of amoxicillin was demonstrated in incurred fish samples containing residues at approximately 50-300 ng/g. PMID- 10820078 TI - Determination of potato glycoalkaloids using a liposome immunomigration, liquid phase competition immunoassay. AB - Polyclonal anti-solanine antibodies were raised and used in the assembly of a liposome immunomigration, liquid-phase competition strip immunoassay. In this format, a similar cross-reactivity was observed between the glycoalkaloids alpha solanine and alpha-chaconine. The strip assay was implemented to quantitate total glycoalkaloids (TGA) from potatoes. Recoveries in spiked potato samples using the strip assay and water:acetic acid:sodium bisulfite extracting solvent were in the range of 84-111%. The values of the TGA quantitations by the strip assay as compared with those obtained by HPLC, in nonspiked tubers coming from cloned potato samples donated by potato breeders, were equivalent and highly correlated (r(2) = 0.91). The strip assay proved advantageous over HPLC for extra-laboratory measurements such as the rapid identification of samples that should be rejected due to an elevated TGA content. PMID- 10820079 TI - Effect of hydrolysis time on the determination of amino acids in samples of soybean products with ion-exchange chromatography or precolumn derivatization with phenyl isothiocyanate. AB - Accurate determination of amino acid levels in soy products facilitates optimum diet formulation and amino acid supplementation. A study was carried out to investigate the effect of hydrolysis time and method of amino acid measurement on amino acid levels. Correction factors to standardize amino acid levels to 24 h of hydrolysis were also determined. Six different soybean products were evaluated. Hydrolysis was carried out for 10 different periods of time. Amino acids were analyzed by both ion-exchange chromatography and precolumn derivatization with phenyl isothiocyanate. Both hydrolysis time and measurement method affected (P < 0.05) amino acid levels. Standard hydrolysis conditions (hydrolysis in 6 M HCl at 110 degrees C for 24 h) rarely provide the maximal amino acid values. Therefore, sequential hydrolyses curves were very useful and should be used. PMID- 10820080 TI - Measuring asparagine synthetase activity in crude plant extracts. AB - Asparagine synthetase B (AS) is the primary enzyme responsible for asparagine synthesis in plants. Routine biochemical studies of this enzyme's activity have been hindered by several problems including enzyme instability and rapid physiological turnover, endogenous inhibitors, competing pathways, and asparaginase activity. We describe an extraction procedure and assay conditions that provide a reliable, direct assay for the determination of AS activity in crude plant extracts. This assay performed well with several leguminous species and the enzyme preparation retained activity for up to 3 weeks when stored at -80 degrees C. Radio-HPLC detection enabled quantitative measurement of de novo aspargine synthesis in the extracts. Optimal activity was obtained with 1 mM glutamine and 10 mM ATP in the reaction assay. Aminooxyacetic acid (AOA, 1 mM) which prevents the assimilation of aspartate into the TCA cycle, was necessary to measure AS activity in peas, but not in lupine or soybean. PMID- 10820081 TI - Correlation of lycopene measured by HPLC with the L, a, b color readings of a hydroponic tomato and the relationship of maturity with color and lycopene content. AB - Tomatoes (Lycopersicon esculentum cv. Laura) were separated, according to the ripening stage, by a sensory panel into seven groups, and color was measured on the tomato surface with a Minolta Chroma meter. The L, a, b, hue, chroma, and lycopene content were plotted against the maturity stages of the tomatoes, and several good correlations were found. The a/b ratio and the lycopene content were the parameters that allowed six of seven maturity groups in the tomato to be statistically distinguished. The lycopene content, measured by HPLC, was also correlated with the color measurements, and the a, a/b, and (a/b)(2) color factors produced the best regressions. An estimation of the lycopene content in tomatoes can be achieved by using a portable chroma meter, with a possible field usage application. Equations to calculate the lycopene content of tomatoes based on the color readings are reported. PMID- 10820082 TI - Identification of flavone C-glycosides including a new flavonoid chromophore from barley leaves (Hordeum vulgare L.) by improved NMR techniques. AB - Six flavone C-glycosides were isolated from young leaves of barley. One of the C glucosides has a new type of nucleus, a 2',4',5,5', 7-penta-OH-substituted flavone bearing a 6-C-beta-D-glucoside, which has apparently never been isolated before. One mono- and two di-C-glycosyl flavones were isolated for the first time from barley and identified as isoscoparin 7-O-beta-D-glucoside, carlinoside, and shaftoside, respectively. Other flavones were 7-O-beta-D-glucosides of isoorientin and isovitexin. The known problematic NMR structure elucidation of C glycosyl flavonoids has been solved by using both a temperature close to the freezing point of the solvent (22.5 degrees C in DMSO-d(6)) and a high temperature (70, 90 degrees C) for comparison during NMR measurements. Structural determination of all the compounds was achieved by employing 1D and 2D NMR techniques. PMID- 10820083 TI - High-performance liquid chromatographic method for determination of biogenic amines in feedstuffs, complete feeds, and animal tissues. AB - Numerous methods to analyze biogenic amines in biological materials have been described. A versatile and rapid methodology to analyze these compounds in feedstuffs, complete feeds, and animal tissues, however, has not been reported. The current method was developed to address this need. Biogenic amines in feedstuffs, complete animal feeds, and animal tissues were extracted with 10% trichloroacetic acid, reacted with O-phthaladehyde using high-performance liquid chromatographic employing a cation exchange column. Detection limits were 50 pmol/mL for tyramine, histamine, putrescine, and spermine; 40 pmol/mL for cadaverine; and 25 pmol/mL for spermidine. Extraction efficiency of biogenic amines in feedstuffs, duodenum, liver, ileum + jejunum, and whole shrimp and shrimp hepatopancreas ranged between 99-105, 93-135, 80-85, 65-102, 88-98, and 88 97%, respectively. It can be concluded that the current method can be applied to individual feedstuffs, complete feeds, and animal tissues for the rapid and accurate determination of concentration of biogenic amines. PMID- 10820084 TI - Changes in phytochemical and antioxidant activity of selected pepper cultivars (Capsicum species) as influenced by maturity. AB - The effect of fruit maturation on changes in carotenoids, flavonoids, total soluble reducing equivalents, phenolic acids, ascorbic acid, and antioxidant activity (AOX) in different pepper types (Capsicum annuum, Capsicum frutescens, and Capsicum chinese) was determined. Generally, the concentration of these chemical constituents increased as the peppers reached maturity. Peppers contained high levels of L-ascorbic acid and carotenoids at maturity, contributing 124-338% of the RDA for vitamin C and 0.33-336 RE/100 g of provitamin A activity, respectively. Levels of phenolic acids, capxanthin, and zeaxanthin generally increased during maturation, whereas the level of lutein declined. Flavonoid concentrations varied greatly among the pepper types analyzed and were negatively correlated to AOX under the conditions of the beta-carotene linoleic assay. Model systems were used to aid in understanding the relationship between flavonoids and AOX. Significant increases in AOX were observed in pepper juice models in response to increasing dilution factors and the presence of EDTA, indicating a pro-oxidant effect due to metal ions in the system. In vitro models demonstrated that increasing levels of flavonoids in combination with constant levels of caffeic and ascorbic acid gave a resultant AOX that was either additive of the two compounds or competitive in their ability to scavenge peroxyl radicals. The model systems were in good agreement with the chemical composition of the pepper cultivars and reflected the interactions affecting AOX. More research is needed to understand the complex interactions that occur among various antioxidants present in pepper extracts. PMID- 10820085 TI - Formation of the mutagen IFP in model systems and detection in restaurant meats. AB - Mixtures of the free amino acids, creatine and glucose, were dry-heated to model the potential formation of heterocyclic amines in meats. The formation of the mutagenic amine IFP (determined to be 2-amino-(1,6-dimethylfuro[3,2-e]imidazo[4,5 b])pyridine) was investigated by varying heating time, heating temperature, and precursors. With an optimized mixture of glutamine, creatine, and glucose, heated at 200 degrees C for 60 min, 2 mg of IFP was purified for studies to define its structure. Trideuteriomethyl-IFP was made from trideuteriomethylcreatinine in the model system for use in LC-MS detection of IFP in foods. Analysis of well-done meats purchased from restaurants showed about half to contain IFP at levels from 1.4 to 46 ng/g of cooked meat, demonstrating human exposure to this mutagen. PMID- 10820086 TI - Fermentation of a bacterial cellulose/xylan composite by mixed ruminal microflora: implications for the role of polysaccharide matrix interactions in plant cell wall biodegradability. AB - Growth of the cellulose-synthesizing bacterium Acetobacter xylinum ATCC 53524 in media supplemented with 5% (w/v) glucose and 0.2% (w/v) of a water-soluble, nearly linear xylan from tobacco stalks resulted in the synthesis of a highly crystalline composite having a xylose/glucose ratio ranging from 0.06 to 0.24. The digestion of one composite (88% cellulose/12% xylan) by mixed ruminal microflora displayed kinetics of gas production similar to those of an unassociated mixture of the two components added in a xylan/cellulose ratio similar to that of the composite. The data suggest that intimate association of xylan and cellulose, as is typically found in secondary plant cell walls, does not inhibit the rate of digestion of the component polysaccharides. PMID- 10820087 TI - Bioavailability of iron from purple laver (Porphyra spp.) estimated in a rat hemoglobin regeneration bioassay. AB - Laver belongs to the genius of Porphyra and is the most valuable seaweed in the aqua-culture industry. It contains higher iron than many other plant foods. The bioavailability of iron from laver was evaluated in a rat hemoglobin regeneration assay. Reagent-grade ferrous sulfate was used as the reference standard, and the relative biological value (RBV) for laver was expressed as a percentage of the response to ferrous sulfate. RBV was calculated by two methods: slope-ratio and ratio of hemoglobin regeneration efficiency, and both yielded RBV of 26 for laver. Amount of available iron from laver estimated from RBV was comparable to many iron-fortified foods. PMID- 10820088 TI - Antioxidant activity and radioprotective effects against chromosomal damage induced in vivo by X-rays of flavan-3-ols (Procyanidins) from grape seeds (Vitis vinifera): comparative study versus other phenolic and organic compounds. AB - The quantitative distribution of several flavan-3-ols was determined using HPLC in a grape (Vitis vinifera) seed extract (GSE) of four cultivars grown in the region of Murcia. Polymer >/= C(4) units made up the largest group of procyanidins in the GSE (90.92%, expressed as HPLC % area). The antioxidant activity of GSE and other reference compounds was investigated by measuring their ability to scavenge the ABTS(*)(+) radical cation (TEAC). The most effective compounds were, in order: GSE > rutin > (+)-catechin > diosmin >/= ascorbic acid. The radioprotective effects of GSE and other reference compounds were determined by using the micronucleus test for anticlastogenic activity, any reduction of the frequency of micronucleated polychromatic erythrocytes (MnPCEs) being evaluated in the bone marrow of mouse exposed to X-rays. The most effective compounds were, in order: GSE > rutin > dimethyl sulfoxide (DMSO) > ascorbic acid > 6-n-propyl-2 thiouracil-6c (PTU) > diosmin. The higher ABTS(*)(+) scavenging capacity and anticlastogenic activity of GSE can be explained, structurally, by the high number of conjugated structures between the catechol groups in the B-rings and the 3-OH free groups of the polymeric polyphenolic skeleton and, in addition, by the stability of the aroxyl flavonoid radical generated in the above processes. PMID- 10820089 TI - Catechin contents of foods commonly consumed in The Netherlands. 1. Fruits, vegetables, staple foods, and processed foods. AB - Catechins, compounds that belong to the flavonoid class, are potentially beneficial to human health. To enable epidemiological evaluation of these compounds, data on their contents in foods are required. HPLC with UV and fluorescence detection was used to determine the levels of (+)-catechin, (-) epicatechin, (+)-gallocatechin (GC), (-)-epigallocatechin (EGC), (-)-epicatechin gallate (ECg), and (-)-epigallocatechin gallate (EGCg) in 24 types of fruits, 27 types of vegetables and legumes, some staple foods, and processed foods commonly consumed in The Netherlands. Most fruits, chocolate, and some legumes contained catechins. Levels varied to a large extent: from 4.5 mg/kg in kiwi fruit to 610 mg/kg in black chocolate. (+)-Catechin and (-)-epicatechin were the predominant catechins; GC, EGC, and ECg were detected in some foods, but none of the foods contained EGCg. The data reported here provide a base for the epidemiological evaluation of the effect of catechins on the risk for chronic diseases. PMID- 10820090 TI - Catechin contents of foods commonly consumed in The Netherlands. 2. Tea, wine, fruit juices, and chocolate milk. AB - Catechins, compounds that belong to the flavonoid class, are potentially beneficial to human health. To enable an epidemiological evaluation of catechins, data on their contents in foods are required. HPLC with UV and fluorescence detection was used to determine the levels of (+)-catechin, (-)-epicatechin, (+) gallocatechin (GC), (-)-epigallocatechin (EGC), (-)-epicatechin gallate (ECg), and (-)-epigallocatechin gallate (EGCg) in 8 types of black tea, 18 types of red and white wines, apple juice, grape juice, iced tea, beer, chocolate milk, and coffee. Tea infusions contained high levels of catechins (102-418 mg of total catechins/L), and tea was the only beverage that contained GC, EGC, ECg, and EGCg in addition to (+)-catechin and (-)-epicatechin. Catechin concentrations were still substantial in red wine (27-96 mg/L), but low to negligible amounts were found in white wine, commercially available fruit juices, iced tea, and chocolate milk. Catechins were absent from beer and coffee. The data reported here provide a base for the epidemiological evaluation of the effect of catechins on the risk for chronic diseases. PMID- 10820091 TI - Insoluble condensed tannins of canola/rapeseed. AB - The contents of soluble, SDS-extractable, and insoluble condensed tannins were determined in canola/rapeseed hulls from several varieties by utilizing the proanthocyanidin assay. The total amount of tannins in rapeseed/canola hulls ranged from 1913 to 6213 mg per 100 g of oil-free hulls. Insoluble tannins predominated in canola/rapeseed hulls and comprised from 70 to 95.8% of total tannins present. The amounts of SDS-extractable tannins were comparable to those of soluble tannins but constituted only 4.7-14. 1% of insoluble tannins present. PMID- 10820092 TI - Evaluation of auraptene content in citrus fruits and their products. AB - Auraptene quantities in Tanaka's 77 Citrus species (including 14 varieties and cultivars), 5 Fortunella species, one Poncirus species, 27 hybrids between Citrus species, and 51 intergeneric hybrids between Citrus and Poncirus have been evaluated. The genus Citrus has been divided into eight groups. Auraptene is found in all of the species of Cephalocitrus group, a part of the species of Aurantium group, and most of the species of Osmocitrus group. The Citrus species contain a small amount of auraptene in the juice sacs compared with in the peels except for Henka mikan (C. pseudo-aurantium), Ichang lemon (C. wilsonii), and a Hassaku (C. hassaku)-pummelo hybrid (Okitsu No. 39), which contain large quantities of auraptene in their juice sacs (0.23, 0.52, and 0.14 mg/g, respectively). The Hong Kong wild kumquat (F. hindusii) alone contains auraptene in Fortunella species. All of the Citrus-trifoliate orange (P. trifoliata) hybrids as well as the trifoliate orange contain a large quantity of auraptene in both the peel (16.57-0.51 mg/g) and the juice sac (10.32-0.15 mg/g). These hybrids are almost inedible. The Iyo (C. iyo)-trifoliate orange hybrid (IyP269) is edible and contains auraptene in the peel (1.49 mg/g) and in the juice sac (1.73 mg/g). Citrus fruit products, for example, brand-named grapefruit juice and marmalade, retain about 0. 1 mg and 0.3 mg/100 g of auraptene, respectively. PMID- 10820093 TI - alpha-tocopherol content of Greek virgin olive oils. AB - alpha-Tocopherol, the main tocopherol homologue found in olive oil, was determined using normal phase HPLC. Ninety Greek virgin olive oils, selected according to a designed sampling protocol from different cultivars and regions all over Greece for three successive crop years, were analyzed. For a specific olive cultivar, which is widely used for the production of olive oil in Greece, additional measurements were made to study the effect of milling conditions on alpha-tocopherol concentration. Finally, a significant number of commercial olive oil samples (25) obtained from the retail market were analyzed. High concentrations of alpha-tocopherol were observed in most of the samples selected from various regions. Values ranging between 98 and 370 mg/kg were found (>200 mg/kg in 60% of samples). Extraction conditions were not found to influence alpha tocopherol level. alpha-Tocopherol content of retail market samples was high, ranging from 120 to 250 mg/kg of oil (>180 mg/kg in 60% of samples). Storage of samples under domestic conditions for two years showed that good handling is quite important for retaining high alpha-tocopherol levels and for increasing, thus, the storage life and nutritional value of this exquisite oil. PMID- 10820094 TI - Comparison of the most odor-active compounds in fresh and dried hop cones (Humulus lupulus L. variety spalter select) based on GC-olfactometry and odor dilution techniques. AB - Application of aroma extract dilution analysis on the volatiles obtained from dried cones of Spalter Select hops grown in the German hop-growing area of Hallertau revealed 23 odorants in the flavor dilution (FD) factor range of 16 4096, 20 of which could be identified. On the basis of high FD factors, trans-4, 5-epoxy-(E)-2-decenal, linalool, and myrcene were identified as the most potent odorants, followed by ethyl 2-methylpropanoate, methyl 2-methylbutanoate, (Z)-1,5 octadien-3-one, nonanal, (E,Z)-1,3, 5-undecatriene, 1,3(E),5(Z),9-undecatetraene, propyl 2-methylbutanoate, 4-ethenyl-2-methoxyphenol, and 1-octen-3-one. Ten of the high-impact hop aroma compounds had previously not been identified as hop constituents and, in particular, 1,3(E),5(Z), 9-undecatetraene has not yet been reported as a food odorant. In an extract obtained from fresh hops, in addition to the odorants found in dry hops, (Z)-3-hexenal was characterized as a further key odorant rendering an additional green aroma note to the fresh material. PMID- 10820095 TI - Interactions between anthocyanins and aroma substances in a model system. Effect on the flavor of grape-derived beverages. AB - Evaluation of the sensory quality of wine or grape-derived beverages led us to study the interactions between flavors and anthocyanins, the colored family of polyphenols. The flavylium cation-ligand complexation, resulting in copigmentation (rise in pigment visible absorption with a concomitant bathochromic shift), was investigated using visible absorption spectroscopy. Sole volatile phenols were found to markedly interact with malvidin-3,5-O-diglucoside. With series of guaiacyl-derived aroma substances, acyl-substituted ligands proved to be better copigments than alkyl-substituted ones. Association constants and 1:1 complex stoichiometry were further determined for several substrates. Decreasing binding to malvin was observed for acetosyringone, syringaldehyde, acetovanillone, vanillin, 3,5-dimethoxyphenol, and 4-ethylguaiacol. Addition of 10% ethanol lowered by one-third the association constants for malvin-ligand couples and for malvidin-3-O-glucoside with acetosyringone and syringaldehyde. The main driving force was ascribed to hydrophobicity, although this study evidenced an influence of the ligand substitution pattern on copigmentation. PMID- 10820096 TI - Influence of a Saccharomyces cerevisiae selected strain in the volatile composition of rose wines. Evolution during fermentation. AB - There has been considerable controversy about the use of selected pure strains in wine fermentation. For that reason it is important to determine the influence of this vinification technique in the composition of wine because it arises from the type of yeast and the subsequent evolution during fermentation. This study researches the volatile composition of rose wines from the Garnacha must, inoculated with one selected NA33 strain of Saccharomyces cerevisiae. The inoculated yeast did not predominate in all of the samples. These samples showed a behavior intermediate between those of the control and samples in which NA33 did predominate. The greatest concentration of higher alcohols was in the control wine, and its evolution was similar in all fermentations. The esters formed at the end of the fermentation and their concentrations were higher in the control than in the inoculated samples. In the control, acids were produced above all, in the first half of fermentation, and decreased from then onward. In the sample in which the yeast predominated, the synthesis occurred later and to a lesser extent than in the control. PMID- 10820097 TI - A powerful aromatic volatile thiol, 2-furanmethanethiol, exhibiting roast coffee aroma in wines made from several Vitis vinifera grape varieties. AB - The chemical compound 2-furanmethanethiol (2FM), with a strong roast coffee aroma, has been identified in sweet white wines made from the Petit manseng grape variety, and in certain red Bordeaux wines (made from the Merlot, Cabernet franc, and Cabernet sauvignon grape varieties). This was done by extracting specific volatile thiols using p-hydroxymercuribenzoate. The 2FM has also been found in toasted oak used in barrel-making. All the Petit manseng sweet white wines and some of the red Bordeaux wines analyzed contained between a few ng/L and several dozen ng/L of 2FM. Taking into account its very low perception threshold (0.4 ng/L in a model hydro alcoholic environment), 2FM could therefore contribute to the roast coffee aroma of certain wines. PMID- 10820098 TI - Volatile flavor components in red fermented soybean (Glycine max) curds. AB - Red fermented soybean (Glycine max) curds (FSC) were extracted with a simultaneous steam distillation and extraction (SDE) apparatus and analyzed by gas chromatography-mass spectrometry (GC-MS) and further evaluated by gas chromatography-mass spectrometry-flame ionization detector-olfactometry (GC-MS FID-O). A combined total of 89 compounds were identified from three samples (A C). The numbers of components identified (samples A-C) were 87, 85, and 81, respectively, which were divided into 10 compound classes. The quantity of 48 compounds was statistically significant (p < 0.05). Sample B generally had the lowest level, whereas sample C had the higher level of each common component among the samples. GC-MS-FID-O analyses showed that the majority of the fruity, diacetyl, cantaloupe-like odors were eluted early during the run (RI < 1100), followed by the meaty flavor, rosy, and prune-like aromas (RI > 1100). Ethyl 2 methylpropanoate, 2,3-butanedione, ethyl butanoate, ethyl 2-methylbutanoate, 3 (methylthio)propanal, benzeneacetaldehyde, and ethyl 3-phenylpropionate were found to be important common components contributing to the characteristic aroma of red FSCs. PMID- 10820099 TI - Flavor release from salad dressings: sensory and physicochemical approaches in relation with the structure. AB - The effect of process and formulation on sensory perception and flavor release was investigated on salad dressing models. Oil/vinegar emulsions (phi = 0.5, droplet size > 10 microm) with thickeners and a whey protein concentrate were prepared with different fat droplet sizes and different distributions of fat droplet size. The effect of the amount of emulsifier was also tested. Sensory profile analysis was performed by a trained panel and flavor release quantified by dynamic headspace analysis. When the droplet size is increased, the lemon smell and citrus aroma significantly increase, whereas the egg note, mustard, and butter aroma significantly decrease. The concentrations of alcohols and acids significantly increase when droplet size increases, whereas those of other compounds such as limonene or benzaldehyde significantly decrease. The dispersion of the droplet size has a small effect on flavor perception, and the effect of the increase of the amount of emulsifier is noticed only by instrumental analysis. PMID- 10820100 TI - Effects of thermal processing and storage on available lysine and furfural compounds contents of infant formulas. AB - The Maillard reaction-related effects that thermal treatments during the manufacturing process and storage (at 20 and 37 degrees C) have on powdered adapted and follow-up milk-based infant formulas were estimated by measuring the available lysine and furfural compounds contents of raw cow milk used in manufacturing, intermediate products and formulas. A fluorimetric method was used to measure the available lysine contents, and free and total furfural compounds were determined by HPLC. Statistically significant losses in available lysine (about 20%) in the infant formulas with respect to raw milk were found. The storage period did not affect the available lysine contents of adapted formulas but reduced (16%) the contents of the follow-up ones (from 6.61 to 5.33 g/100 g of protein). No furfural compounds were detected in raw milk, and free and total furyl methyl ketone (FMC) and methylfurfural (MF) were not observed in the analyzed samples. After 6 months of storage, an increase in free hydroxymethylfurfural (HMF) (from 0.34 to 0.77 mg/100 g of protein) and furfural (F) (from nondetectable to 0.1 mg/100 g of protein) in adapted formulas and free HMF (from 1.84 to 2.62 mg/100 g of protein) in follow-up formulas was observed. PMID- 10820101 TI - Kinetics of formation of Di-D-fructose dianhydrides during thermal treatment of inulin. AB - Thermal treatments of solid mixtures of inulin and citric acid result in the formation of di-D-fructose dianhydrides and oligomers derived therefrom. The kinetics of formation of these compounds have been investigated and simulated in computer studies. A mechanism is proposed. The conditions used in this study were analogous to the conditions pertaining to the roasting of chicory, during which similar compounds are formed. PMID- 10820102 TI - Volatile changes during dehydration of d'Agen prunes. AB - Solid phase microextraction (SPME) was used in conjunction with GC-MS to monitor the changes in some major volatile flavors during drying of plums. The aroma profile was significantly modified during the process and substantial loss of the original volatile flavors was observed. The generation of some compounds was shown to be due to the thermal decomposition of carbohydrates. This paper discusses the importance of aroma profiling in detecting the progress of the chemical (degradation) reactions and identifying marker volatiles in quality control of the product. PMID- 10820103 TI - Oxidation of encapsulated oil in tailor-made cellular solid. AB - A cellular alginate solid containing oil was prepared by freeze-drying. The oil was incorporated in the matrix by emulsification in the pre-gel state. The alginate-oil gels were immersed in 60 degrees Brix sucrose solution for various periods, before freeze-drying. The extent of the collapse expressing the reduction in sample volume was affected by immersion duration and freeze-drying conditions. Sucrose diffusion during immersion followed an exponential pattern. Effective diffusivity calculated using nonlinear regression gave a value of 3.64 x 10(-)(10) m(2)/s. The effect of relative humidity on water content calculated on a dry basis excluding sucrose showed a significant increase in water content at 75% RH. Image analysis was utilized to quantify the area of the encapsulated oil droplets. The area of the droplets was divided into four subregions defined as (0.02-0.1) x 10(-)(12), (0. 1-1.0) x 10(-)(12), (1-10) x 10(-)(12), and (10 100) x 10(-)(12) m(2). A distribution resembling a Gaussian bell distribution with a maximum of 54% for the (1-10) x 10(-)(12) m(2) area range was found. The number of oil droplets was almost constant for the first three area regions, and then decreased markedly. Oxidation index was not effected by porosity at 0 and 22% RH. A 75% RH and porosity above a critical value of ca. 0.45 was found to increase oxidation significantly. Samples immersed for less than 5.5 h in sucrose solution were mechanically stronger after equilibration at 0 and 22% RH when compared to their counterpart equilibrated at 75% RH. Immersion for more than 24 h resulted in similar mechanical strength irrespective of the RH. PMID- 10820104 TI - Lipoxygenase inactivation in green beans (Phaseolus vulgaris L.) due to high pressure treatment at subzero and elevated temperatures. AB - The kinetics of lipoxygenase (LOX) inactivation in green beans due to high pressure treatment were studied in the pressure-temperature area of 0.1 up to 650 MPa and -10 up to 70 degrees C for systems with different levels of food complexity, i.e., in green bean juice and intact green beans (in situ study). For both systems, LOX was irreversibly inactivated by high-pressure treatment combined with subzero and elevated temperatures and the inactivation could be described as a first-order reaction. At ambient pressure, in situ LOX was less thermostable than in the juice at temperatures below 68 degrees C whereas the stability ranking was reverse at temperatures above 68 degrees C. At temperatures below 63 degrees C, sensitivity of the inactivation rate constants to temperature changes was on the same order of magnitude in the juice and in situ, while at higher temperature it was lower in situ. The pressure needed to obtain the same rate of LOX inactivation at a given temperature was lower in situ than in the juice. Application of high-pressure treatment at low/subzero temperature resulted in an antagonistic effect on LOX inactivation for both systems, whereas no such effect was found above room temperature. The pressure-temperature dependence of the LOX inactivation rate constants in green beans was successfully modeled. PMID- 10820105 TI - Natural occurrence of fumonisins in corn from Iran. AB - Corn collected in the Mazandaran and Isfahan Provinces of Iran was analyzed for fumonisin B(1) (FB(1)), fumonisin B(2) (FB(2)), and fumonisin B(3) (FB(3)). The samples from Mazandaran Province, situated on the Caspian littoral of Iran, were random samples from farmers' corn lots collected in September 1998, whereas those from Isfahan Province, situated further south in the center of Iran, were bought as corn cobs in the local retail market during October 1998. All 11 samples from Mazandaran showed high levels of fumonisin contamination with FB(1) levels between 1.270 and 3.980 microg/g, FB(2) levels between 0.190 and 1.175 microg/g, and FB(3) levels between 0.155 and 0.960 microg/g. Samples from Isfahan showed lower levels of contamination with eight of eight samples having detectable FB(1) (0.010-0.590 microg/g), two of eight samples having detectable FB(2) (0.050-0.075 microg/g), and two of eight samples having detectable FB(3) (0.050-0.075 microg/g). This is the first report of fumonisin contamination of corn from Iran, in which samples from the area of high esophageal cancer on the Caspian littoral have been shown to contain high levels of fumonisins. PMID- 10820106 TI - Influence of halogen salts on the production of the ochratoxins by Aspergillus ochraceus Wilh. AB - The first report of the biological production of bromo ochratoxin B by Aspergillus ochraceus Wilh. is presented as well as a study of the influence of potassium bromide, potassium iodide, potassium fluoride, and potassium chloride on the production of ochratoxin A and ochratoxin B. Potassium fluoride and potassium iodide inhibited the growth of the fungus, whereas potassium chloride substantially stimulated the production of ochratoxin A in shaken solid substrate fermentation on whole wheat or shredded wheat, generally giving a high yield of ochratoxins. Increasing levels of potassium bromide led to a decline in ochratoxin A production and an increase in bromo-ochratoxin B, ochratoxin B, and 4-hydroxy ochratoxin B. Nevertheless, A. ochraceus was much less versatile in the bromo analogues than other fungi, which produce metabolites containing chlorine. Analysis included aminopropyl solid-phase extraction column cleanup followed by quantitative analysis on reversed-phase HPLC using fluorescence detection and employing N-(5-chloro-2-hydroxybenzoyl)phenylalanine as an internal standard. PMID- 10820107 TI - C(18) acetylenic fatty acids of Ximenia americana with potential pesticidal activity. AB - Bioactivity-driven fractionation of the CHCl(3) extract of the root of Ximenia americana, using the brine shrimp lethality test (BST) and hatchability test with Clavigralla tomentosicollis eggs, gave C(18) acetylenic fatty acids 1 and 2. 1 is octadeca-5-ynoic acid (tariric acid). 2 is a novel ene-ene-yne-ene acetylenic fatty acid (10Z,14E,16E-octadeca-10,14,16-triene-12-ynoi c acid). The structures of 1 and 2 were assigned from the MS and NMR data. Fractions that are rich in acetylenic fatty acids inhibited the hatching of C. tomentosicollis eggs. PMID- 10820108 TI - Structure elucidation and antifungal activity of an anthracycline antibiotic, daunomycin, isolated from Actinomadura roseola. AB - The actinomycete strain Ao108 producing antifungal metabolites active against some plant pathogenic fungi was identified as Actinomadura roseola, based on the analyses of morphological and physiological characteristics. The antibiotic Da2B that showed a strong antifungal activity was isolated from the culture broth and mycelial mats of A. roseola strain Ao108 using various chromatographic procedures. On the basis of (1)H NMR, (13)C NMR, and 2-D NMR correlation data, the antibiotic Da2B was confirmed to have the structure of an anthracycline antibiotic, daunomycin. In vitro antimicrobial spectrum tests showed that the antibiotic Da2B had substantial inhibitory activity (10 microg mL(-)(1) of MICs) against mycelial growth of Phytophthora capsici and Rhizoctonia solani. The antibiotic also showed antiyeast activity against Saccharomyces cerevisiae, but the growth of Candida albicans was not affected. Antibacterial activity was found only against Gram-positive bacteria. In the further evaluation of in vivo efficacy, application of the antibiotic Da2B effectively inhibited the development of Phytophthora blight in pepper plants. However, the control efficacy of the antibiotic against Phytophthora infection was somewhat less than that of metalaxyl. The antibiotic Da2B did not show any phytotoxicity on pepper plants even at 500 microg mL(-)(1). PMID- 10820109 TI - Convenient synthesis of 2,4-diacetylphloroglucinol, a natural antibiotic involved in the control of take-all disease of wheat. AB - 2,4-Diacetylphloroglucinol (DAPG) is an antibiotic with broad-spectrum antibacterial and antifungal activities. It is a major determinant in the biological control of several plant diseases. DAPG is produced by Pseudomonas fluorescens both in vitro and in the rhizosphere of wheat. It is involved in the natural suppression of take-all disease known as take-all decline, which develops in soils following extended monoculture of wheat or barley. A one-step synthesis of DAPG from the commercially available 2-acetylphloroglucinol is described. This reaction involves the direct alkylation of 2-acetylphloroglucinol using acetic anhydride as the acetylation reagent, with boron trifluoride-etherate as the catalyst. This synthesis is simple and produces higher yields of DAPG (90%) as compared with previously described procedures. As ecological concerns are gaining equal status with agricultural concerns, the demand for natural biocontrol measures is increasing. There is tremendous pressure from society on agriculture to reduce the use of pesticides. A discussion is given on the agricultural and ecological importance of this natural antibiotic and its application as an alternative to reduce the use of synthetic pesticides. PMID- 10820110 TI - Insect antifeedant flavonoids from Gnaphalium affine D. Don. AB - The antifeedant flavonoids, 5-hydroxy-3,6,7,8,4'-pentamethoxyflavone (1), 5 hydroxy-3,6,7,8-tetramethoxyflavone (2), 5,6-dihydroxy-3, 7-dimethoxyflavone (3), and 4,4',6'-trihydroxy-2'-methoxychalcone (4), have been isolated from cudweed Gnaphalium affine D. Don (Compositae). Four natural flavonoids showed insect antifeedant activity against the common cutworm (Spodoptera litura F.). These flavonoids were detected in small amounts in the plant by HPLC analysis, but these natural compounds had strong antifeedant activity against the common cutworm. On the other hand, 4 was detected in a large amount in the plant, but this compound had only a slight activity. Therefore, these natural compounds were regarded as one of the plant's defensive systems against phytophagous insects along with the woolly plant surface. As for the structure-activity relationship, it is an advantage for antifeedant activity to have no oxy-substituents on the B ring of the flavonoid but have an ether linkage such as a pyran in the chemical structure. PMID- 10820111 TI - Development and testing of seven new synthetic coyote attractants. AB - Available evidence indicates that effective coyote attractants are blends of volatile substances. Typically, attractants are a combination of biological substances such as fermented glandular materials, urines, and rotted meats. Although effective, these attractants have several distinct disadvantages. Among these is the possibility that they are unnecessarily complex and variable and, thus, difficult to replicate from one batch to the next. Although attractants containing a few reagent grade materials are available, the chemicals selected and their concentrations are not derived from actual attractants. For this reason, commercially available coyote attractants were analyzed with the intention of developing relatively simple synthetic alternatives. Purge and trap headspace analysis with gas chromatography/mass selective detection was employed to identify the volatile components of known conventional and synthetic attractants. All identified compounds were grouped according to chemical functionality, and one compound from each functional group was chosen to represent the group. Using only these representative compounds, seven synthetic attractants were formulated. Bioassays with captive coyotes (Canis latrans) were conducted to compare behavioral responses elicited by the seven new attractants, a currently available synthetic attractant, and a control. The results indicated that the attractants elicited significantly different behavioral profiles. PMID- 10820112 TI - Factors affecting the dissolution and degradation of oriental mustard-derived sinigrin and allyl isothiocyanate in aqueous media. AB - Sinigrin, the predominant glucosinolate in the oriental mustard Brassica juncea, is mainly degraded upon the enzymatic action of myrosinase under normal conditions to give allyl isothiocyanate (AITC) in an aqueous media. Because AITC is considered to be the principal nematicidal ingredient in B. juncea, its stability in aqueous media is an important issue in achieving efficient nematode control. Pure sinigrin and AITC were found to be relatively stable in buffered water in the pH range of 5.00-7.00 but less stable at pH 9.00. Both sinigrin and AITC were more stable in soil water (supernatant of a 1:1 water/air-dried soil mixture) than in buffered water at the same pH range of 5.00-9.00. Sinigrin dissolved from the mustard bran or ground seed into water very quickly and was degraded by codissolved myrosinase to AITC. The AITC that formed from the degradation of sinigrin was found to be more stable in the soil water than in the buffered water. Buffer capacity was considered to be one of the factors that contributed to the stabilization of AITC in the soil water, but other unknown factors from both bran or seed and soil may also have contributed to the stabilization. PMID- 10820113 TI - Growth inhibitory effects on fall armyworm Spodoptera frugiperda of some limonoids isolated from Cedrela spp. (Meliaceae). AB - Dichloromethane extracts of Cedrela salvadorensis and Cedrela dugessi afforded a photogedunin epimeric mixture, gedunin and cedrelanolide. These compounds and the photogedunin epimeric acetates 3 and 4 at the 23-OH position were evaluated against Spodoptera frugiperda. Toosendanin, isolated from Melia azedarach, was used as a positive control. When tested for activity on neonate larvae into the no-choice bioassays, gedunin, photogedunin epimeric mixture, and photogedunin acetates mixture caused significant larval mortality with LC(50) of 39.0, 10.0, and 8.0 ppm at 7 days, respectively, as well as growth reduction. All the compounds tested inhibited larval growth, compared to the control, in a concentration-dependent manner. In addition, it was possible to observe significant reduced pupal weights and adult emergence. All the tested compounds except cedrelanolide showed comparable activity to that of toosendanin. PMID- 10820114 TI - Chlordane uptake and its translocation in food crops. AB - Chlordane is a member of the persistent organic pollutants (POPs), a group of chemicals characterized by extremely long residence in the environment after application. Technical chlordane, composed of a large number of components, is a synthetic organochlorine substance that was used primarily as an insecticide. Uptake by root crops of persistent soil residues of chlordane was noted early in the chronology of the material. The present report is the first comprehensive study of the uptake of weathered soil residues of chlordane and its translocation throughout the tissues of food crops under both greenhouse and field conditions. The data show that for all 12 crops chlordane is not limited to root tissue but is translocated from the root to some of the aerial tissues. Chlordane accumulation in edible aerial tissue appears to be dependent on plant physiology. As expected, chlordane was detected in the edible root tissue of the three root crops examined, carrots, beets, and potatoes. In the remaining crops chlordane was detected in the edible aerial tissue of spinach, lettuce, dandelion, and zucchini, whereas it was not detected in edible aerial tissue of tomatoes, peppers, and corn; trace amounts of chlordane were detected in the edible aerial tissue of bush beans and eggplant. Under the conditions of the field trial the data indicate that for weathered chlordane residues, the soil-to-plant uptake route dominates over the air-to-plant uptake route. This is the case even when the soil concentration of the recalcitrant, weathered residues, for which volatilization is expected to be minimal, is as high as it would be directly following application. Greenhouse trials confirm this observation for zucchini, a member of the Cucurbitaceae family, which bioaccumulates weathered chlordane very efficiently in its edible fruits. PMID- 10820115 TI - Use of stable tracer studies to evaluate pesticide photolysis at elevated temperatures. AB - New methods were developed to determine photolysis rates of medium-weight pesticides in the gas phase using elevated air temperatures and solid-phase microextraction (SPME). A 57-L glass chamber was constructed that utilized collimated xenon arc irradiation that could heat chamber air to increase the amount of pesticide in the gas phase. Gas-phase photolysis rates were determined at various air temperatures by comparing the rate of loss of each of the tested pesticides to a photochemically stable tracer, hexachlorobenzene. Interval sampling of gas-phase constituents was performed using SPME immediately followed by GC-ECD or GC-MSD analysis. The two pesticides under examination were the dinitroaniline herbicide trifluralin and the organophosphorus insecticide chlorpyrifos. The gas-phase photolysis for trifluralin was found to be rapid with half-lives of 22-24 min corrected for sunlight. These results were comparable to photochemical lifetime estimates from other investigators under sunlight conditions. Elevating temperatures from 60 to 80 degrees C did not affect photolysis rates, and these rates could be extrapolated to environmental temperatures. From 60 to 80 degrees C, gas-phase chlorpyrifos photolysis lifetimes were observed to range from 1.4 to 2.2 h corrected for sunlight and will thus be important together with hydroxyl radical reactions for removing this substance from the atmosphere. At these elevated temperatures, pesticides and tracer compounds were found to be substantially in the gas phase, and possible effects on reaction rates from wall interactions were minimized. PMID- 10820116 TI - Reactivity of certain organophosphorus insecticides toward hydroxyl radicals at elevated air temperatures. AB - Methods were developed to determine OH reaction rates for medium-weight organophosphorus pesticides in the gas phase. A 57-L chamber was constructed that utilized xenon arc irradiation (>290 nm) to photolyze the OH precursor, methyl nitrite. Experiments were performed at elevated temperatures ranging from 60 to 80 degrees C to minimize wall sorption. Solid-phase microextraction (SPME) sampling of the gas phase was employed to assess the simultaneous rate of loss of the pesticides in relation to the rate of loss of two reference substances with known OH rate constants. An internal gas-phase standard (hexachlorobenzene), relatively stable to hydroxyl radicals, was used to assess other processes, which included dilution and wall sorption. The relative reaction rates of the organophosphorus insecticides, diazinon and chlorpyrifos, when compared to reference compounds, were unaffected by various air temperatures between 60 and 80 degrees C. Although both insecticides were expected to react at similar rates on the basis of structural activity model predictions, diazinon reacted 3 times more rapidly than chlorpyrifos and gave estimated environmental OH half-lives of 0.5 and 1.4 h, respectively. The degree of sorption onto the chamber walls was minimal and similar for each of the compounds examined. Experimental gas-phase determinations at elevated temperatures may provide important information that can be used when in the assessment of the potential of airborne pesticide risks to nontarget and ecologically sensitive areas. PMID- 10820117 TI - Influence of soil pH-sorption interactions on imazethapyr carry-over. AB - Soil pH affects imazethapyr sorption-desorption, which in turn can affect persistence and bioavailability. Long-term imazethapyr carry-over has been observed in soil that is below pH 6.5, resulting in significant sugarbeet damage. Imazethapyr concentration decreased rapidly in field soil, regardless of pH. Despite similar amounts of imazethapyr remaining in aged soils at different pH levels, there were differences in bioavailability, which can be explained by sorption-desorption. At low pH more imazethapyr was sorbed than at high pH, but it readily desorbed. At high pH less imazethapyr was sorbed initially, but it did not readily desorb. Thus, after 3 months, the remaining imazethapyr in low-pH soil was desorbable and bioavailable, resulting in injury to canola and sugarbeet. Liming aged, low-pH soil released bound imazethapyr residues, which would then be degraded and result in less carry-over. PMID- 10820118 TI - Sorption and catalytic hydrolysis of diethatyl-ethyl on homoionic clays. AB - Sorption and catalytic hydrolysis of the herbicide diethatyl-ethyl [N chloroacetyl-N-(2,6-diethylphenyl)glycine ethyl ester] on homoionic Na(+)-, K(+) , Ca(2+)-, and Mg(2+)-montmorillonite clays were studied in aqueous media. The Freundlich sorption coefficient, K(f), measured from isotherms on clay followed the order of Na(+) approximately K(+) > Mg(2+) approximately Ca (2+). Analysis of FT-IR spectra of diethatyl-ethyl sorbed on clay suggests probable bonding at the carboxyl and amide carbonyl groups of the herbicide. The rate of herbicide hydrolysis in homoionic clay suspensions followed the same order as that for sorption, indicating that sorption may have preceded and thus caused hydrolysis. Preliminary product identification showed that hydrolysis occurred via nucleophilic substitution at the carboxyl carbon, causing cleavage of the ester bond and formation of diethatyl and its dechlorinated derivative, and at the amide carbon, yielding an ethyl ester derivative and its acid. These pathways also suggest that hydrolysis of diethatyl-ethyl was catalyzed by sorption on the clay surface. PMID- 10820119 TI - Bioremediation of olive oil mill wastewater: chemical alterations induced by Azotobacter vinelandii. AB - An environmentally friendly bioremediation system of olive oil mill wastewater (OMWW) is studied with respect to its physicochemical characteristics and degradation efficiency on major characteristic constituents. The method exploits the biochemical versatility of the dinitrogen fixing bacterium Azotobacter vinelandii (strain A) to grow in OMWW at the expense of its constituents and to transform it into an organic liquid fertilizer. The system eliminates the phytotoxic principles from OMWW and concomitantly enriches it with an agriculturally beneficial microbial consortium along with useful metabolites of the latter. The end product, branded "biofertilizer", is used as soil conditioner and liquid organic fertilizer. Growth of A. vinelandii in OMWW results in the decline of content of most of the compounds associated with phytotoxicity, and this is confirmed by the assessment of degradation yields. In parallel, during the process several other compounds noncommittally undergo degradation and biotransformation. More specifically, the biofertilization system is capable of achieving removal yields as high as 90 and 96% after 3 and 7 days of treatment, respectively. Statistical analysis of the results showed that between the periods of operation no significant difference occurs with respect to the degradation yield. Moreover, the degradation yield from 3 to 7 days of continuous operation of the system remains almost unaltered during 2 consecutive years. PMID- 10820120 TI - Characterization of different commercial soybean peroxidase preparations and use of the enzyme for N-demethylation of methyl N-methylanthranilate To produce the food flavor methylanthranilate. AB - The potential of different peroxidase preparations for the N-demethylation of methyl N-methylanthranilate to produce the food flavor methylanthranilate (MA) was investigated. All tested peroxidase preparations were able to catalyze the N dealkylation. The tested soybean preparations vary widely with respect to their heme content. Furthermore, the operational stability of purified soybean peroxidase (SP) is at least 25-fold lower than that of horseradish peroxidase and only 5-fold higher than that of microperoxidase 8. Thus, the presence of a large protein chain around a porphyrin cofactor in a peroxidase is, by itself, insufficient to explain the observed differences in operational stability. Despite its relatively low operational stability, SP proved to be the most efficient biocatalyst for the production of MA with high yield and purity, especially observed at the high temperature and low pH values at which SP appeared to be optimally active. PMID- 10820121 TI - Novel approach for large-scale, biocompatible, and low-cost fractionation of peptides in proteolytic digest of food protein based on the amphoteric nature of peptides. AB - A large-scale, biocompatible, and low-cost procedure for peptide fractionation based on the amphoteric nature of peptide is developed. A sample cell (120 x 100 x 50 mm) with four joint tubes (17 mm i.d. and 20 mm in length) on the front and back was prepared. On the end of the joint tubes, a nylon screen (100 mesh) supported agarose gel layer was formed. Five or nine of the sample cells were connected. A tryptic digest of casein (2.0-3.6 L) was applied to the sample cells. At each end of the sample cell apparatus, an additional cell filled with 0.1 M H(3)PO(4) or NaOH was connected and used as anode and cathode compartments, respectively. Reproducible fractionation of peptide could be achieved by collecting fractions with specific pH values when voltage reached a plateau by applying direct current at constant power. Running time necessary for fractionation of peptide was inversely proportional to electric power and directly proportional to sample volume. PMID- 10820122 TI - Inactivation of orange pectinesterase by combined high-pressure and -temperature treatments: a kinetic study. AB - Pressure and/or temperature inactivation of orange pectinesterase (PE) was investigated. Thermal inactivation showed a biphasic behavior, indicating the presence of labile and stable fractions of the enzyme. In a first part, the inactivation of the labile fraction was studied in detail. The combined pressure temperature inactivation of the labile fraction was studied in the pressure range 0.1-900 MPa combined with temperatures from 15 to 65 degrees C. Inactivation in the pressure-temperature domain specified could be accurately described by a first-order fractional conversion model, estimating the inactivation rate constant of the labile fraction and the remaining activity of the stable fraction. Pressure and temperature dependence of the inactivation rate constants of the labile fraction was quantified using the Eyring and Arrhenius relations, respectively. By replacing in the latter equation the pressure-dependent parameters (E(a), k(ref)(T)()) by mathematical expressions, a global model was formulated. This mathematical model could accurately predict the inactivation rate constant of the labile fraction of orange PE as a function of pressure and temperature. In a second part, the stable fraction was studied in more detail. The stable fraction inactivated at temperatures exceeding 75 degrees C. Acidification (pH 3.7) enhanced thermal inactivation of the stable fraction, whereas addition of Ca(2+) ions (1 M) suppressed inactivation. At elevated pressure (up to 900 MPa), an antagonistic effect of pressure and temperature on the inactivation of the stable fraction was observed. The antagonistic effect was more pronounced in the presence of a 1 M CaCl(2) solution as compared to the inactivation in water, whereas it was less pronounced for the inactivation in acid medium. PMID- 10820123 TI - Optimization of simultaneous saccharification and fermentation for the production of ethanol from lignocellulosic biomass. AB - Simultaneous saccharification and fermentation (SSF) of alkaline hydrogen peroxide pretreated Antigonum leptopus (Linn) leaves to ethanol was optimized using cellulase from Trichoderma reesei QM-9414 (Celluclast from Novo) and Saccharomyces cerevisiae NRRL-Y-132 cells. Response surface methodology (RSM) and a three-level four-variable design were employed to evaluate the effects of SSF process variables such as cellulase concentration (20-100 FPU/g of substrate), substrate concentration (5-15% w/v), incubation time (24-72 h), and temperature (35-45 degrees C) on ethanol production efficiency. Cellulase and substrate concentrations were found to be the most significant variables. The optimum conditions arrived at are as follows: cellulase = 100 FPU/g of substrate, substrate = 15% (w/v), incubation time = 57.2 h, and temperature = 38.5 degrees C. At these conditions, the predicted ethanol yield was 3.02% (w/v) and the actual experimental value was 3.0% (w/v). PMID- 10820124 TI - Development of a polyclonal antibody-based sensitive enzyme-linked immunosorbent assay for fumonisin B(4). AB - Polyclonal antibodies (PAb) against fumonisin B(4) (FmB(4)), which have good cross-reactivity with four major fumonisins, were produced by immunizing a rabbit with FmB(4)-keyhole limpet hemocyanin conjugate. A sensitive competitive direct enzyme-linked immunosorbent assay (CD-ELISA) for fumonisins was developed. Because of the limited supply of FmB(4), both FmB(1)-horseradish peroxidase conjugate (HRP) and FmB(3)-HRP were tested as the toxin-enzyme markers in the CD ELISA. In the FmB(1)-HRP-based CD-ELISA, the concentrations of FmB(1), FmB(2), FmB(3), and FmB(4) causing 50% inhibition of binding of enzyme marker (IC(50)) were 9.0, 2.1, 9.0, and 6.5 ng/mL (or the relative cross-reactivities toward FmB(1), FmB(2), FmB(3), and FmB(4) were 58.5, 309.5, 58.5, and 100%), respectively. In the FmB(3)-HRP-based CD-ELISA, the IC(50) values for FmB(1), FmB(2), FmB(3), and FmB(4) were 7.1, 1.9, 7.6, and 5.3 ng/mL (or the relative cross-reactivities toward FmB(1), FmB(2), FmB(3), and FmB(4) were 74, 280, 70, and 100%), respectively. The FmB(3)-HRP-based CD-ELISA was then used in a series of analytical recovery experiments using Fusarium moniliforme corn culture material spiked with FmB(1) and with clean corn spiked with a FmB(3)/FmB(4) containing extract. The overall recovery of FmB(1) from culture material in the range of 10-100 ppm was 65%. The detection limit for FmB(1) with clean corn as matrix was between 100 and 500 ppb. F. moniliforme cultures were analyzed with the developed CD-ELISA and a well-established FmB(1) antibody-based ELISA, which is not sensitive to FmB(4). Differences in the fumonisin levels found by the two assays were used as an indication of the presence of FmB(4) in the culture material and, therefore, as a method to identify FmB(4)-producing strains. Using ELISA in combination with HPLC individual B-series fumonisins were quantified. The ELISA developed in the present study would be a useful supplement to FmB(1) antibody-based ELISA for screening of Fusarium strains for the production of major fumonisins. PMID- 10820125 TI - Mapping of spinocerebellar ataxia 13 to chromosome 19q13.3-q13.4 in a family with autosomal dominant cerebellar ataxia and mental retardation. AB - We examined a large French family with autosomal dominant cerebellar ataxia (ADCA) that was excluded from all previously identified spinocerebellar ataxia genes and loci. The patients-seven women and a 4-year-old boy-exhibited slowly progressive childhood-onset cerebellar gait ataxia associated with cerebellar dysarthria, moderate mental retardation (IQ 62-76), and mild developmental delays in motor acquisition. Nystagmus and pyramidal signs were also observed in some cases. This unique association of clinical features clearly distinguishes this new entity from other previously described ADCA. Cerebral magnetic-resonance imaging showed moderate cerebellar and pontine atrophy in two patients. We performed a genomewide search and found significant evidence for linkage to chromosome 19q13.3-q13.4, in an approximately 8-cM interval between markers D19S219 and D19S553. PMID- 10820126 TI - Testing the robustness of the new Haseman-Elston quantitative-trait loci-mapping procedure. AB - Variance components (VC) techniques have emerged as among the more powerful methods for detection of quantitative-trait loci (QTL) in linkage analysis. Allison et al. found that, with particularly marked leptokurtosis in the phenotypic distribution and moderate-to-high residual sibling correlation, maximum likelihood (ML) VC methods may produce a severe excess of type I errors. The new Haseman-Elston (NHE) method is a least-squares-based VC method for mapping of QTL in sib pairs (Elston et al.). Using simulation, we investigate the robustness of the NHE to marked nonnormality, by means of the same distributions and worst-case conditions identified by Allison et al. for the ML approach (i.e., 100 pairs; high residual sibling correlation). Results showed that, when marked nonnormality is present, the NHE can be used without severe type I error-rate inflation, even at very small alpha levels. PMID- 10820127 TI - A genomic scan of families with prostate cancer identifies multiple regions of interest. AB - A 10-cM genomewide scan of 94 families with hereditary prostate cancer, including 432 affected men, was used to identify regions of putative prostate cancer susceptibility loci. There was an average of 3.6 affected, genotyped men per family, and an overall mean age at diagnosis of 65.4 years. A total of 50 families were classified as early onset (mean age at diagnosis <66 years), and 44 families were classified as later onset (mean age at diagnosis > or =66 years). When the entire data set is considered, regions of interest (LOD score > or =1.5) were identified on chromosomes 10, 12, and 14, with a dominant model of inheritance. Under a recessive model LOD scores > or =1.5 were found on chromosomes 1, 8, 10, and 16. Stratification by age at diagnosis highlighted a putative susceptibility locus on chromosome 11, among the later-onset families, with a LOD score of 3. 02 (recombination fraction 0) at marker ATA34E08. Overall, this genomic scan suggests that there are multiple prostate cancer loci responsible for the hereditary form of this common and complex disease and that stratification by a variety of factors will be required for identification of all relevant genes. PMID- 10820128 TI - High incidence of propionic acidemia in greenland is due to a prevalent mutation, 1540insCCC, in the gene for the beta-subunit of propionyl CoA carboxylase. AB - Propionyl CoA carboxylase (PCC) is a mitochondrial, biotin-dependent enzyme involved in the catabolism of amino acids, odd-chain fatty acids, and other metabolites. PCC consists of two subunits, alpha and beta, encoded by the PCCA and PCCB genes, respectively. Inherited PCC deficiency due to mutations in either gene results in propionic acidemia (PA), an autosomal recessive disease. Surprisingly, PA is highly prevalent among Inuits in Greenland. We have analyzed reverse transcriptase-PCR products of the beta-subunit mRNA, to characterize the responsible mutation(s). A 3-bp insertion, 1540insCCC, was found in homozygous form in three patients and in compound heterozygous form in one patient. The resulting PCC has no measurable activity, and the mutant beta-subunit appears to be very unstable. To test the hypothesis that a common mutation is responsible for PA in the Greenlandic Inuit population, 310 anonymous DNA samples of Inuit origin were screened for 1540insCCC. We found a carrier frequency of 5%, which is very high compared with those of most other autosomal recessive diseases. Analysis of alleles of a very closely linked marker, D3S2453, revealed a high degree of linkage disequilibrium between one specific allele and 1540insCCC, suggesting that this mutation may be a founder mutation. PMID- 10820131 TI - Short communication. The effects of drinking and smoking on the CYP2D6 metabolic capacity. AB - We studied the influence of drinking and smoking habits on CYP2D6 metabolic capacity measured by the use of debrisoquine as a substance test. We did not find any significant differences in the frequency of subjects with CYP2D6 deficiency (poor metabolizers) among four groups of healthy individuals: nonsmokers/nondrinkers, smokers/drinkers, nondrinkers/smokers, and nonsmokers/drinkers. We demonstrated that, among poor metabolizers, alcohol and tobacco consumption was associated with higher metabolic ratios than it was with the control group, but the differences were not statistically significant. Among extensive metabolizers, the lowest metabolic ratio (highest enzyme activity) was detected for nondrinkers/smokers, intermediate values for smokers/drinkers, and the highest metabolic ratio (lowest enzyme activity) for nonsmokers/drinkers. These variations were slight but statistically significant when logarithmic ratio values were applied. These results show that smoking and drinking habits do not need to be taken into account when humans are phenotyped for CYP2D6. PMID- 10820129 TI - Localization of a recessive gene for North American Indian childhood cirrhosis to chromosome region 16q22-and identification of a shared haplotype. AB - North American Indian childhood cirrhosis (NAIC, or CIRH1A) is an isolated nonsyndromic form of familial cholestasis reported in Ojibway-Cree children and young adults in northwestern Quebec. The pattern of transmission is consistent with an autosomal recessive mode of inheritance. To map the NAIC locus, we performed a genomewide scan on three DNA pools of samples from 13 patients, 16 unaffected siblings, and 22 parents from five families. Analysis of 333 highly polymorphic markers revealed 3 markers with apparent excess allele sharing among affected individuals. Additional mapping identified a chromosome 16q segment shared by all affected individuals. When the program FASTLINK/LINKAGE was used and a completely penetrant autosomal recessive mode of inheritance was assumed, a maximum LOD score of 4.44 was observed for a recombination fraction of 0, with marker D16S3067. A five-marker haplotype (D16S3067, D16S752, D16S2624, D16S3025, and D16S3106) spanning 4.9 cM was shared by all patients. These results provide significant evidence of linkage for a candidate gene on chromosome 16q22. PMID- 10820130 TI - Evidence for a prostate cancer-susceptibility locus on chromosome 20. AB - Recent studies suggest that hereditary prostate cancer is a complex disease involving multiple susceptibility genes and variable phenotypic expression. While conducting a genomewide search on 162 North American families with > or =3 members affected with prostate cancer (PRCA), we found evidence for linkage to chromosome 20q13 with two-point parametric LOD scores >1 at multiple sites, with the highest two-point LOD score of 2.69 for marker D20S196. The maximum multipoint NPL score for the entire data set was 3.02 (P=.002) at D20S887. On the basis of findings from previous reports, families were stratified by the presence (n=116) or absence (n=46) of male-to-male transmission, average age of diagnosis (<66 years, n=73; > or =66 years, n=89), and number of affected individuals (<5, n=101; > or =5, n=61) for further analysis. The strongest evidence of linkage was evident with the pedigrees having <5 family members affected with prostate cancer (multipoint NPL 3.22, P=.00079), a later average age of diagnosis (multipoint NPL 3.40, P=.0006), and no male-to-male transmission (multipoint NPL 3.94, P=.00007). The group of patients having all three of these characteristics (n=19) had a multipoint NPL score of 3.69 (P=.0001). These results demonstrate evidence for a PRCA susceptibility locus in a subset of families that is distinct from the groups more likely to be linked to previously identified loci. PMID- 10820132 TI - Ethylphenidate formation in human subjects after the administration of a single dose of methylphenidate and ethanol. AB - Ethylphenidate was recently reported as a novel drug metabolite in two overdose fatalities where there was evidence of methylphenidate and ethanol coingestion. This study explores the pharmacokinetics of ethylphenidate relative to methylphenidate and the major metabolite ritalinic acid, in six healthy subjects who received methylphenidate and ethanol under controlled conditions. Subjects (three males, three females) received a single oral dose of methylphenidate (20 mg; two 10-mg tablets) followed by consumption of ethanol (0.6 g/kg) 30 min later. Methylphenidate, ritalinic acid, and ethylphenidate were quantified using liquid chromatography-tandem mass spectrometry. Ethylphenidate was detectable in the plasma and urine of all subjects after ethanol ingestion. The mean (+/-S.D.) area under the concentration versus time curve for ethylphenidate was 1.2 +/- 0.7 ng/ml/h, representing 2.3 +/- 1.3% that of methylphenidate (48 +/- 12 ng/ml/h). A significant correlation was observed between the area under the concentration versus time curve of methylphenidate and that of ethylphenidate. In view of the known dopaminergic activity of racemic ethylphenidate, it remains possible that under certain circumstances of higher level dosing, e.g., in the abuse of methylphenidate and ethanol, the metabolite ethylphenidate may contribute to drug effects. PMID- 10820133 TI - Immunohistochemical localization of the acylases that catalyze the deacetylation of N-acetyl-L-cysteine and haloalkene-derived mercapturates. AB - Acylases catalyze the hydrolysis of a range of S-substituted N-acetyl-L cysteines. The hydrolysis of N-acetyl-L-cysteine is catalyzed by cytosolic acylase I, and activity is present in human endothelial cells and rat lung, intestinal, and liver homogenates. Many haloalkenes are metabolized to mercapturates, which also undergo acylase-catalyzed hydrolysis. The acylases that catalyze the deacetylation of N-acetyl-L-cysteine and several haloalkene-derived mercapturates have been recently identified: acylase I catalyzes the deacetylation of N-acetyl-L-cysteine and some haloalkene-derived mercapturates whereas an acylase purified from rat kidney cytosol catalyzes the deacetylation of a distinct set of substrates, including several haloalkene-derived mercapturates. The objective of these studies was to examine the tissue and subcellular localization of acylase I and purified rat kidney acylase. Immunoblotting showed the presence of immunoreactive acylase I and purified rat kidney acylase in rat kidney, liver, lung, and brain. Both acylases were identified by immunohistochemistry in several rat organs, including kidney, liver, lung, brain, stomach, intestines, adrenals, pancreas, and testis, indicating that acylase activity is widespread in rat tissues. PMID- 10820134 TI - Metabolism of A dopamine D(4)-selective antagonist in rat, monkey, and humans: formation of A novel mercapturic acid adduct. AB - 3-([4-(4-Chlorophenyl)piperazin-1-yl]-methyl)-1H-pyrrolo-2, 3-beta-pyridine (L 745,870) is a dopamine D(4) selective antagonist that has been studied as a potential treatment for schizophrenia, with the expectation that it would not exhibit the extrapyramidal side effects often observed with the use of classical antipsychotic agents. The metabolism of L-745,870 in vivo was investigated in the rat, rhesus monkey, and human using liquid chromatography-tandem mass spectrometry and/or NMR techniques in conjunction with radiochemical detection. In all three species, two major metabolic pathways were identified, namely N dealkylation at the substituted piperazine moiety and the formation of a novel mercapturic acid adduct. It is proposed that the latter biotransformation process involves the formation of an electrophilic imine methide intermediate, analogous to that produced from 3-methyl indole. This report appears to represent the first example of metabolic activation of a 3-alkyl-7-azaindole nucleus. PMID- 10820135 TI - A distribution study with (14)C-otilonium bromide in the rat: evidence for selective tropism for large intestine after oral administration. AB - The aim of this study was to determine the plasma levels and the tissue distribution of otilonium bromide, measured as total radioactivity, after oral administration of 2 mg/kg of (14)C-labeled drug to rats. Radioactivity levels were very low in the plasma (ranging from 2.7 ng Eq/ml at 1.5 h to 0.6 ng Eq/ml at 24 h) as compared with those found in the gastrointestinal (GI) tract, indicating negligible systemic otilonium bromide absorption. Results from both quantitative radioluminography of whole body tissue distribution and radioassay of dissected parts of the GI tract carried out with liquid scintillation counting clearly demonstrate the presence of radioactive compounds in the walls of the GI tract at all sacrifice times. In the other tissues and organs examined, radioactivity was only found in trace amounts in the liver. The presence of radioactivity in the GI walls reflected the transit kinetics of drug-enriched contents. The radioactivity in large intestine walls was measurable at otilonium bromide concentrations in the range of micromole equivalents/kg, from 4 to 8 h after drug administration. Total body radioactivity recovery was 95, 101, 24, and 9% at 1.5, 4, 8, and 24 h, respectively. In conclusion, orally administered (14)C otilonium bromide is poorly absorbed systemically, as indicated by the very low plasma radioactivity levels, but it is able to effectively penetrate into the large intestine walls, a recognized target for drugs oriented toward irritable bowel syndrome therapy. PMID- 10820136 TI - Metabolism of dipropyl disulfide by rat liver phase I and phase II enzymes and by isolated perfused rat liver. AB - The metabolism of dipropyl disulfide (DPDS), an Allium sulfur compound, was investigated in rat liver cell subfractions and in an isolated perfused rat liver. DPDS is oxidized to dipropyl thiosulfinate (DPDSO) by rat microsomes. The contribution of cytochrome P450 enzymes (CYPs) or flavin-containing monooxygenases (FMO) to the formation of DPDSO from its precursor was investigated. In rat microsomes, the reaction followed Michaelis-Menten kinetics with a K(m) = 0.52 +/- 0.1 mM and a V(max) = 5.91 +/- 0.5 nmol/min/mg of protein, respectively (mean +/- S.E., n = 4). Both FMOs and CYPs were involved in DPDS oxidation, although the contribution of CYPs was predominant. Liver microsomes from phenobarbital-treated rats showed a 3.2-fold increase in the rate of formation of DPDSO. Among many CYP isoform-specific inhibitors, only CYP2B1/2 inhibitors decreased the formation of DPDSO and the best correlation between the rate of DPDS oxidation with specific monooxygenase activities was found with a marker of CYP2B1/2 activity. The action of phase II enzymes on DPDS metabolism was studied by incubating DPDS or DPDSO with liver cytosols or microsomes. Two metabolites were formed from DPDS: propylglutathione sulfide conjugate and propylmercaptan, whereas with DPDSO, only the glutathione conjugate was observed. No conjugate compound was detected in the presence of UDP-glucuronyl transferases. When isolated rat liver was perfused with DPDS, different metabolites were obtained in the output and in the liver tissues: propylmercaptan appeared in the output, whereas methylpropyl sulfide, methylpropyl sulfone, and propylglutathione sulfide were detected in the liver tissue. PMID- 10820137 TI - Pharmacological inhibition of P-glycoprotein transport enhances the distribution of HIV-1 protease inhibitors into brain and testes. AB - HIV protease inhibitors have proven remarkably effective in treating HIV-1 infection. However, some tissues such as the brain and testes (sanctuary sites) are possibly protected from exposure to HIV protease inhibitors due to drug entry being limited by the membrane efflux transporter P-glycoprotein, located in the capillary endothelium. Intravenous administration of the novel and potent P glycoprotein inhibitor LY-335979 to mice (1-50 mg/kg) increased brain and testes concentration of [(14)C]nelfinavir, up to 37- and 4-fold, respectively, in a dose dependent fashion. Similar effects in brain levels were also observed with (14)C labeled amprenavir, indinavir, and saquinavir. Because [(14)C]nelfinavir plasma drug levels were only modestly increased by LY-335979, the increase in brain/plasma and testes/plasma ratios of 14- to 17- and 2- to 5-fold, respectively, was due to increased tissue penetration. Less potent P-glycoprotein inhibitors like valspodar (PSC-833), cyclosporin A, and ketoconazole, as well as quinidine and verapamil, had modest or little effect on brain/plasma ratios but increased plasma nelfinavir concentrations due to inhibition of CYP3A-mediated metabolism. Collectively, these findings provide "proof-of-concept" for increasing HIV protease inhibitor distribution into pharmacologic sanctuary sites by targeted inhibition of P-glycoprotein using selective and potent agents and suggest a new therapeutic strategy to reduce HIV-1 viral replication. PMID- 10820138 TI - The glucuronidation of morphine by dog liver microsomes: identification of morphine-6-O-glucuronide. AB - Canines are used extensively in the pharmaceutical industry for the preclinical screening of novel therapeutics, yet comparatively little is known about the phase 2 metabolism in this species. In humans, morphine is known to undergo extensive metabolism by glucuronidation, and the UDP-glucuronosyltransferase isoform, which catalyzes the formation of morphine-3-O-glucuronide and morphine-6 O-glucuronide is UGT2B7. This study was designed to investigate the glucuronidation of morphine using dog liver microsomes. Liver microsomes from beagle dogs catalyzed the glucuronidation of morphine-3(and 6)-O-glucuronide at rates 4 to 10 times that of rhesus monkey and human liver microsomes. The K(m) of morphine using beagle dog liver microsomes was approximately 270 microM, which is similar to that found for expressed human UGT2B7. The V(max) for morphine, using dog liver microsomes, was 27 nmol/min/mg of protein. Flunitrazepam inhibited the glucuronidation of morphine in dog liver microsomes, and the K(i) was 40 microM, which is similar to human UGT2B7 for other substrates. The effects of detergents were also investigated with dog liver microsomes, and Brij 35 and Brij 58 were found to be the best detergents to use for maximal activation of the dog liver morphine UGT. These studies suggest that dog has a UGT2B isoform similar to human UGT2B7. PMID- 10820139 TI - In vitro inhibition and induction of human hepatic cytochrome P450 enzymes by modafinil. AB - The ability of modafinil to affect human hepatic cytochrome P450 (CYP) activities was examined in vitro. The potential for inhibition of CYP1A2, CYP2A6, CYP2B6, CYP2C9, CYP2C19, CYP2D6, CYP2E1, CYP3A4/5, and CYP4A9/11 by modafinil (5-250 microM) was evaluated with pooled human liver microsomes. Modafinil exhibited minimal capacity to inhibit any CYP enzyme, except CYP2C19. Modafinil inhibited the 4'-hydroxylation of S-mephenytoin, a marker substrate for CYP2C19, reversibly and competitively with a K(i) value of 39 microM, which approximates the steady state C(max) value of modafinil in human plasma at a dosage of 400 mg/day. No irreversible inhibition of any CYP enzyme was observed, and there was no evidence of metabolism-dependent inhibition. The potential for induction of CYP activity was evaluated by exposing primary cultures of human hepatocytes to modafinil (10 300 microM). Microsomes were then prepared and assayed for CYP1A2, CYP2A6, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, and CYP3A4/5 activities. The mean activities of microsomal CYP1A2, CYP2B6, and CYP3A4/5 from modafinil-treated hepatocytes were higher (up to 2-fold) than those in the solvent-treated controls but were less than those produced by reference inducers of these enzymes. At high concentrations of modafinil (>/=100 microM), the mean activity of CYP2C9 was decreased (up to 60%) relative to that in the solvent controls. Overall, modafinil was shown to have effects on human hepatic CYP1A2, CYP2B6, CYP2C9, CYP2C19, and CYP3A4/5 activities in vitro. Although effects obtained in vitro are not always predictive of effects in vivo, such results provide a rational basis for understanding drug-drug interactions that are observed clinically and for planning subsequent investigations. PMID- 10820140 TI - First-pass disposition of (-)-6-aminocarbovir in rats: II. Inhibition of intestinal first-pass metabolism. AB - A CBV [(-)-carbovir, (-)-carbocyclic 2',3'-didehydro-2', 3'-dideoxyguanosine] prodrug, 6AC [(-)-6-aminocarbovir, (-)-carbocyclic 2',3'-didehydro-2', 3'-dideoxy 6-deoxy-6-aminoguanosine], was previously evaluated in rats, and it exhibited superiority to the parent drug in increasing systemic and central nervous system exposure to CBV. The gut wall was determined to be the dominant site of the first pass activation of 6AC after lumenal administration. If subsequent delivery to the brain is desired, then such a first-pass effect might not be viewed favorably. Because the first-pass conversion of 6AC primarily takes place in the intestine by adenosine deaminase (ADA), quenching of the intestinal activation of 6AC by oral administration of ADA inhibitors may result in an increased 6AC bioavailability, and thus an improved brain exposure to CBV. The objectives of the study were to determine whether the ADA inhibitors 2'-deoxycoformycin and erythro-9-(2-hydroxy-3-nonyl)adenine were capable of achieving a substantial and selective inhibition of gut wall activation of 6AC, and to determine whether the systemic concentrations of 6AC would be thus increased. Thirty-nine male Sprague Dawley rats were divided into two groups. One group received 6AC by either the portal vein or intralumenally with the coadministration of intralumenal 2' deoxycoformycin. Similarly, the other group received 6AC with coadministration of erythro-9-(2-hydroxy-3-nonyl)adenine. Substantial suppression of the first-pass conversion of 6AC was achieved with both inhibitors. This inhibition appeared to be relatively selective, allowing the choice of dose of inhibitor that would sufficiently inhibit the first-pass metabolism while leaving the activation capacity in the systemic circulation unaltered. The systemic level of 6AC increased with the escalating dose of inhibitors, thus increasing the driving force for passive uptake into the brain. PMID- 10820141 TI - Comparison of imidazole- and 2-methyl imidazole-containing farnesyl-protein transferase inhibitors: interaction with and metabolism by rat hepatic cytochrome P450s. AB - Methylation at the 2-position of the imidazole ring of IBN (I), a 1, 5 substituted imidazole-containing compound, was carried out to minimize its inhibition of rat cytochrome P450 (CYP)3A activity. The resulting analog 2-MIBN (II) exhibited an inhibitory potency 70-fold weaker (K(i) = 25 microM) than that of I (K(i) = 0.3 microM) toward CYP3A, the major rat liver microsomal P450 isoform(s) for the metabolism of I and II by rat liver microsomes in the presence of NADPH. The structural modification did not switch the major metabolic pathways for I and II, but significantly decreased the affinity of II to the metabolizing enzyme(s) as reflected by the difference in their K(i) values for CYP3A. Enzyme kinetic studies also demonstrated that I had a lower apparent K(m) (0.3 microM) than than II (18 microM), but an apparent V(max) 14 times lower than II. This finding indicates that methylation at the imidazole ring reduced the affinity of the compound to CYP3A, but increased the catalytic capacity, turning I as a substrate of low K(m) value but low capacity into a compound of high K(m) but high capacity for the metabolism. Our results suggest the significance of substrate concentration in comparing the metabolic stability of compounds with different kinetic parameters. Although higher intrinsic clearance is implied for I when the substrate concentration is below or close to its K(m) value, higher metabolic rate was constantly seen with II over micromolar range. The different kinetic parameters of I and II may also explain the observation that no significant difference in pharmacokinetic behavior was seen after an i.v. administration of I and II to the rat. PMID- 10820142 TI - Pharmacokinetics and biological fate of 3-(2,2, 2-trimethylhydrazinium)propionate dihydrate (MET-88), a novel cardioprotective agent, in rats. AB - In this study, we examined the disposition, metabolism, and excretion of a novel cardioprotective agent, 3-(2,2, 2-trimethylhydrazinium)propionate dihydrate (MET 88), in rats. The disposition of MET-88 after oral and i.v. administration of 2, 20, and 60 mg/kg indicated that the pharmacokinetics of MET-88 were nonlinear. The profiles of radioactive MET-88 and total radioactivity in plasma were consistent at doses of 20 and 60 mg/kg. However, at 2 mg/kg, the plasma MET-88 levels were obviously lower than the total. The excretion of radioactivity after oral administration of MET-88 indicated that increasing doses led to a shift from exhaled CO(2) to urinary excretion as the major excretion route. Major metabolites in plasma after oral administration of MET-88 were glucose, succinic acid, and 3-hydroxypropionic acid, and in vitro studies revealed that MET-88 was converted to 3-hydroxypropionic acid by gamma-butyrobetaine hydroxylase (EC 1.14. 11.1). An isolated liver perfusion system modified to trap CO(2) gas was used to examine the excretion pathway of MET-88. [(14)C]CO(2) gas was decreased by the addition of iodoacetic acid, DL-fluorocitric acid, or gamma-butyrobetaine to this system, and subsequent thin-layer chromatography analyses of perfusates revealed that MET-88 was first converted to 3-hydroxypropionic acid by gamma-butyrobetaine hydroxylase and then was biosynthesized to glucose and metabolized to CO(2) gas via the glycolytic pathway and tricarboxylic acid cycle. PMID- 10820143 TI - Metabolism of sameridine to monocarboxylated products by hepatocytes isolated from the male rat. AB - The metabolism of sameridine (LPB) (an amide-type local anesthetic-analgesic agent with a hexyl side chain) to carboxylic acid derivatives by isolated male rat hepatocytes was studied using gradient reversed-phase HPLC and mass spectrometry. Incubation of sameridine with hepatocytes resulted in the formation of numerous different metabolites. Two carboxylic acids, i.e., the C(6) and C(4) carboxylated derivatives of sameridine (LPB-6'-oic acid and LPB-4'-oic acid), were found to be produced from the intermediate omega-hydroxy metabolite (6' hydroxy-LPB). Shortening of the alkyl chain in LPB-6'-oic acid by two carbon atoms resulted in LPB-4'-oic acid. However, incubation of rat hepatocytes with 5' hydroxy-LPB [the (omega-1)-hydroxy derivative of sameridine] did not give rise to any carboxylated derivative. Addition of SKF525A inhibited the metabolism of sameridine by rat hepatocytes, indicating that the initial step is catalyzed by cytochrome P450. Furthermore, the metabolism of sameridine to LPB-4'-oic acid was enhanced in hepatocytes isolated from rats treated with clofibrate, an up regulator of peroxisomal fatty acid beta-oxidation and of microsomal cytochrome P450 4A. L-Carnitine (which increases the rate of mitochondrial fatty acid beta oxidation) had no effect on the level of LPB-4'-oic acid produced by isolated rat hepatocytes. The metabolism of 6'-hydroxy-LPB to LPB-6'-oic acid was inhibited almost completely by 4-methylpyrazole, an inhibitor of alcohol dehydrogenase. Considered together, our findings suggest that cytochrome P450 4A, cytosolic dehydrogenases, and the enzymes involved in peroxisomal fatty acid beta-oxidation catalyze the metabolism of sameridine to LPB-4'-oic acid. PMID- 10820145 TI - Use of a novel real-time quantitative reverse transcription-polymerase chain reaction method to study the effects of cytokines on cytochrome P450 mRNA expression in mouse liver. AB - In this study, we developed a real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) method to study cytochrome P450 (CYP) mRNA regulation by cytokines in mouse liver. The method combines standard RT-PCR with a fluorogenic probe in which the intensity of fluorescence is proportional to the amount of target template present. We show that this method provides very rapid, sensitive, and reproducible quantification of mouse Cyp mRNA with a wide dynamic range of starting target molecule. We validated the method by using several prototypic CYP inducers. As expected, the mRNA levels of Cyp3a11, Cyp2b10, and Cyp1a2 were induced by a single dose of dexamethasone (100 mg/kg), phenobarbital (80 mg/kg), and 3-methylcholanthrene (80 mg/kg), respectively. The method of real time RT-PCR was then used to evaluate the effects of interleukin (IL)-6 (100 ng/mouse), IL-1beta (500 ng/mouse), and tumor necrosis factor (TNF)-alpha (2 microgram/mouse) on Cyp mRNA expression in the mouse. Constitutive Cyp2b10 mRNA was reduced to 40% by IL-6 and 15% by IL-1beta, whereas Cyp2d9 mRNA was reduced to 70% by TNF-alpha administration. The level of Cyp1a2 mRNA was suppressed to 67% by IL-6 and 59% by TNF-alpha. Cyp3a11 and Cyp2e1 mRNAs were not affected by any of the cytokines that were studied. We conclude that the real-time RT-PCR method is a powerful new tool to study CYP mRNA expression and regulation. Using this method, we are the first to report that the expression of constitutive Cyp2b10, 1a2, and 2d9 mRNAs was suppressed by proinflammatory cytokines. PMID- 10820144 TI - Dose and inducer-dependent induction of cytochrome P450 1A in endothelia of the eel, including in the swimbladder rete mirabile, a model microvascular structure. AB - Endothelium is a common site of cytochrome P450 1A (CYP1A) induction in vertebrates, and endothelial CYP1A could affect the distribution and toxicity of CYP1A substrates. We investigated CYP1A induction in organs rich in endothelium, gill, heart, and a microvascular model, the swimbladder rete mirabile, in the eel. Benzo[a]pyrene (BP) and 3, 3',4,4'-tetrachlorobiphenyl (TCB), radiolabeled and injected intraperitoneally, showed similar distribution in eels, with dose dependent increases in concentration in heart and rete mirabile. BP [given at 0.1, 1, and 10 mg/kg (0.4, 4, and 40 micromol/kg)], TCB [given at 0.1, 1, and 10 mg/kg (0.3, 3, 30, and 60 micromol/kg)], and beta-naphthoflavone (BNF) [given at 0.1, 1, 5, 10, and 100 mg/kg (0.4, 4, 20, 40, and 400 micromol/kg)] induced microsomal CYP1A and ethoxyresorufin O-deethylase in heart and rete mirabile. Immunohistochemical analysis confirmed that induction of CYP1A in heart and rete mirabile occurs in the endothelium. Increasing doses of each compound caused increasing penetration of induction into the vascular bed of the rete, but with BNF and BP induction penetrated further than with TCB. At high doses of BNF there also was induction in epithelial cells adjacent to endothelium in gill and kidney. CYP1A also was induced in heart and rete mirabile of eels from sites heavily contaminated by aryl hydrocarbon receptor (AHR) agonists. The penetration of CYP1A induction into capillaries of the rete mirabile reflects the penetration of the inducer itself, consistent with the idea that endothelial CYP1A can indicate the local distribution of AHR agonists. The microvascular rete mirabile in the eel provides a model system to explore further a hypothesis that endothelial CYP1A participates in removal of some AHR agonists from the circulation and to examine the consequences of CYP1A induction to the vascular system. PMID- 10820146 TI - Developmentally regulated alternative splicing of the alpha1(XI) collagen chain: spatial and temporal segregation of isoforms in the cartilage of fetal rat long bones. AB - Type XI collagen is a component of the heterotypic collagen fibrils of fetal cartilage and is required to maintain the unusually thin diameter of these fibrils. The mature matrix form of the molecule retains an N-terminal variable region whose structure is modulated by alternative exon splicing that is tissue specific and developmentally regulated. In the alpha1(XI) chain, antibodies to two of the peptides, p6b and p8, encoded by the alternatively spliced exons localized these epitopes to the surface of the collagen fibrils and were used to determine the pattern of isoform expression during the development of rat long bones (humerus). Expression of the p6b isoform was restricted to the periphery of the cartilage underlying the perichondrium of the diaphysis, a pattern that appears de novo at embryonic Day (E) 14. P8 isoforms appeared to be associated with early stages of chondrocyte differentiation and were detected throughout prechondrogenic mesenchyme and immature cartilage. After E16, p8 isoforms gradually disappeared from the diaphysis and then from the epiphysis preceding chondrocyte hypertrophy, but were highly evident at the periarticular joint surface, where ongoing chondrogenesis accompanies the formation of articular cartilage. The spatially restricted and differentiation-specific distribution of alpha1(XI) isoforms is evidence that Type XI collagen participates in skeletal development via a mechanism that may be distinct from regulation of fibrillogenesis. PMID- 10820147 TI - DNA fiber-FISH staining mechanism. AB - Fluorescence in situ hybridization to DNA fibers (Fiber-FISH) is a high resolution, wide-ranging physical DNA mapping method that finds increasing application in the study of pathological gene rearrangements. Here we present experiments designed to understand the nature of the discontinuous FISH signal patterns seen after Fiber-FISH. Use of a novel cisplatin-based chemical labeling method enabled us to produce intact biotin-labeled cosmid target DNA molecules. We monitored by immunofluorescence the fate of such cosmid targets during denaturation and hybridization. The same cosmid DNA labeled with digoxigenin by nick-translation was used to analyze the FISH probe signal distribution in a different color. The probe signals proved to be a subset of the target signals remaining after denaturation and hybridization. We argue that the discontinuity of probe signals in Fiber-FISH is mainly caused by loss of target DNA and limited accessibility due to in situ renaturation and attachment. Furthermore, we conclude that FISH sensitivity is determined by hybridization efficiency and not the ability to generate sufficient signal from small probes. (J Histochem Cytochem 48:743-745, 2000) PMID- 10820148 TI - Distribution of IGFBP-rP1 in normal human tissues. AB - IGFBP-rP1/mac25 is a recently described member of the insulin-like growth factor binding protein (IGFBP) family. It has structural homology to the other members of the IGFBP family but has a lower affinity for insulin-like growth factors (IGFs). In previous studies using RNA blot hybridization, it was shown that the expression of IGFBP-rP1/mac25 was ubiquitous in normal human tissues. In this report we show by immunohistochemistry that the expression of IGFBP-rP1/mac25 is actually restricted to certain organs and specific cell types. We used an antibody raised against a decapeptide of the C-terminal part of the protein that recognizes a approximately 37-kD protein under reduced conditions. The immunohistochemistry performed on normal human tissues showed a ubiquitous intense staining of peripheral nerves and a variable degree of positive staining in smooth muscle cells, including those from blood vessel walls, gut, bladder, and prostate. Cilia from the respiratory system, epididymis, and fallopian tube showed intense immunoreactivity. Most endothelial cells showed some positivity, whereas fat cells, plasma cells, and lymphocytes were negative. There was specific expression limited to certain cell types in the kidney, adrenal gland, and skeletal muscle, indicating a possible specialized function of IGFBP rP1/mac25 in these organs. We further noted an opposite pattern of staining in the lining epithelium of breast (typically positive) and prostate glands (largely negative). The specific localization of IGFBP-rP1/mac25 as described implies a function of the protein. However, its regulation within the IGF axis or a possible direct action of IGFBP-rP1/mac25 remains to be demonstrated. PMID- 10820149 TI - Thymidylate synthase expression in patients with colorectal carcinoma using a polyclonal thymidylate synthase antibody in comparison to the TS 106 monoclonal antibody. AB - Colorectal cancer is one of the most common human cancers, for which 5 fluorouracil (5FU) is usually part of the treatment. Thymidylate synthase (TS), the target enzyme for 5FU, can be predictive for the outcome of 5FU-based therapy. TS levels in tumor samples can be determined with radiochemical enzyme assays, RT-PCR, and immunohistochemical staining. We validated TS immunohistochemistry with a polyclonal rabbit anti-human TS antibody using the avidin-biotin method. This antibody can be used on paraffin-embedded, formalin fixed material using an antigen retrieval method with citrate buffer and microwave treatment. The antibody shows a granular cytosolic staining pattern. The reproducibility in cross-sections from colorectal tumors from 50 patients was 90% and the interobserver variability was acceptable with a kappa of 0.45. On Western blotting it detects purified TS at 36 kD, while in 5FU-treated cells the ternary complex between FdUMP, TS, and 5, 10-methylene-tetrahydrofolate is clearly visible at 38 kD, with no other interfering bands. In a separate set of tumors, immunostaining was compared with enzyme levels; Western blots correlated with enzyme levels. Because both this polyclonal antibody and the monoclonal antibody TS-106 are being used for large-scale studies, we also determined whether they could be used interchangeably. No differences were observed. This polyclonal antibody is specific and gives reproducible results. A study on a larger scale is ongoing to determine the role of TS as a predictive parameter in patients with colorectal cancer treated either with postoperative adjuvant 5FU/levamisole or with surgery only. PMID- 10820150 TI - Expression of pancreatic islet MHC class I, insulin, and ICA 512 tyrosine phosphatase in low-dose streptozotocin-induced diabetes in mice. AB - Activated immune cells contribute to the development of diabetes mellitus in multiple low-dose streptozotocin-treated mice. However, a role in the process for MHC Class I restricted T-cells remains a matter of debate. In this study, we examined by confocal microscopy the pancreatic expression of MHC Class I protein, insulin, and ICA 512 protein tyrosine phosphatase in C57BL/Ks mice given 40 mg/kg bw streptozotocin IP on 5 consecutive days. All animals were hyperglycemic from Day 7 and onwards. A loss of ICA 512 from the central portions of the islets was noted on Day 3. On Day 7, an increase in MHC Class I expression, confined primarily to immune cells in the exocrine pancreas and the periinsular areas, was detected. Later, several MHC class I/glucagon and some MHC class I/insulin double positive cells were found. The insulitis was maximal on Day 14 and declined thereafter. The induction of MHC Class I expression in endocrine cells, occuring only after the cellular infiltration and when the animals were diabetic, indicates that the immune component of the disease does not depend on MHC Class I restricted cytotoxic T-cells but rather comprises a non-antigen-specific process. (J Histochem Cytochem 48:761-767, 2000) PMID- 10820151 TI - Presence of the Kv1.5 K(+) channel in the sinoatrial node. AB - The aim of this study was to establish, using immunolabeling, whether the Kv1.5 K(+) channel is present in the pacemaker of the heart, the sinoatrial (SA) node. In the atrial muscle surrounding the SA node and in the SA node itself (from guinea pig and ferret), Western blotting analysis showed a major band of the expected molecular weight, approximately 64 kD. Confocal microscopy and immunofluorescence labeling showed Kv1.5 labeling clustered in atrial muscle but punctate in the SA node. In atrial muscle, Kv1.5 labeling was closely associated with labeling of Cx43 (gap junction protein) and DPI/II (desmosomal protein), whereas in SA node Kv1.5 labeling was closely associated with labeling of DPI/II but not labeling of Cx43 (absent in the SA node) or Cx45 (another gap junction protein present in the SA node). Electron microscopy and immunogold labeling showed that the Kv1.5 labeling in atrial muscle is preferentially associated with desmosomes rather than gap junctions. PMID- 10820152 TI - Confocal laser scanning microscopy of rat follicle development. AB - This study used confocal laser scanning microscopy (CLSM) to study follicular development in millimeter pieces of rat ovary. To use this technology, it is essential to stain the tissue before laser excitation with the confocal microscope. Various fluorescent stains (Yo-Pro, Bo-Pro, LysoTracker Red, hydroethidine, ethidium bromide, and 7-aminoactinomycin-d) were applied either to fresh tissue or to tissue that had been fixed with glutaraldehyde or paraformaldehyde. After fixation and staining, the tissue was dehydrated with MEOH and cleared with benzyl alcohol/benzyl aldehyde. CLSM was then used with the appropriate laser excitation, dichroics, and bandpass filters to acquire images of oocytes contained in follicles. Analysis of the data revealed three principal findings. First, a rapid increase in oocyte size occurred in the preantral stages of follicle development. In the antral stage of follicle development, there was a rapid increase in follicle size without any substantial increase in oocyte size. Second, accompanying these changes in oocyte and follicle growth was a differential staining pattern in which the nucleus stained more than the cytoplasm in a young follicle, but stained less than the cytoplasm as the follicle enlarged into the late antral stage. Lastly, using CLSM, atretic follicles showed increased LysoTracker Red staining in the granulosa region of the antral follicle, suggestive of cell death. PMID- 10820153 TI - Connexin43 gene expression and regulation in the rodent seminiferous epithelium. AB - Connexin43 (Cx43) is one of the most predominant gap junction proteins found in the testis. We used in situ hybridization and indirect immunofluorescence to study the distribution of Cx43 mRNA and protein in the rodent seminiferous epithelium. During mouse testis maturation, Cx43 mRNA and its corresponding protein were first detected in the adluminal compartment of the growing seminiferous tubules (early postnatal age: Day 12) to become progressively located in the basal compartment at later ages (Days 16, 19, 27). In seminiferous tubules of sexually mature animals, the intensity of the hybridization signal was stage-dependent, with a maximum at Stage VII compared with Stages V and IX of the spermatogenic cycle (p<0.05). The highest expression of Cx43 mRNA was observed in the supporting Sertoli cells and, to a lesser extent, in the most basally located and less mature germ cells (spermatogonia and spermatocytes). Consistent with these observations, in situ dye coupling was observed between Sertoli cells and basal germ cells. In a mutant mouse deficient for the retinoid X receptor beta, which exhibited abnormal spermatogenesis due to altered Sertoli cell function, Cx43 transcripts were markedly decreased in the seminiferous epithelium (p<0.01). The immunoreactive signal for Cx43 was significantly reduced in seminiferous tubules of the 3-month-old mutant mice (p<0.05) and undetectable in older animals. These data provide new information about the precise localization of Cx43 mRNA and protein in seminiferous tubules of immature and mature rodent testes. Moreover, they suggest that retinoids, through the RXRbeta receptors, could be involved in the control of Cx43 gene expression in Sertoli cells. PMID- 10820154 TI - Immunodetection of alpha1E voltage-gated Ca(2+) channel in chromogranin-positive muscle cells of rat heart, and in distal tubules of human kidney. AB - The calcium channel alpha1E subunit was originally cloned from mammalian brain. A new splice variant was recently identified in rat islets of Langerhans and in human kidney by the polymerase chain reaction. The same isoform of alpha1E was detected in rat and guinea pig heart by amplifying indicative cDNA fragments and by immunostaining using peptide-specific antibodies. The apparent molecular size of cardiac alpha1E was determined by SDS-PAGE and immunoblotting (218 +/- 6 kD; n = 3). Compared to alpha1E from stably transfected HEK-293 cells, this is smaller by 28 kD. The distribution of alpha1E in cardiac muscle cells of the conducting system and in the cardiomyoblast cell line H9c2 was compared to the distribution of chromogranin, a marker of neuroendocrine cells, and to the distribution of atrial natriuretic peptide (ANP). In serial sections from atrial and ventricular regions of rat heart, co-localization of alpha1E with ANP was detected in atrium and with chromogranin A/B in Purkinje fibers of the conducting system in both rat atrium and ventricle. The kidney is another organ in which natriuretic peptide hormones are secreted. The detection of alpha1E in the distal tubules of human kidney, where urodilatin is stored and secreted, led to the conclusion that the expression of alpha1E in rat heart and human kidney is linked to regions with endocrine functions and therefore is involved in the Ca(2+)-dependent secretion of peptide hormones such as ANP and urodilatin. PMID- 10820155 TI - Expression of epidermal growth factor and its receptor in cirrhotic liver disease. AB - Polypeptide growth factors, including epidermal growth factor (EGF), play a central role in regulating hepatocyte growth both in vivo and in primary culture. To characterize EGF gene expression in the pathogenesis of regenerative cirrhotic fibrosis, we employed biotinylated antisense oligonucleotide probes to localize hepatic mRNA transcripts in situ. In control tissue and regenerative hepatic nodules, EGF receptor (EGFR) mRNA transcripts were expressed constitutively. In contrast, oligonucleotide probes targeting the human EGF coding region showed that EGF transcription was extremely low in control liver but was highly elevated and localized to regenerative hepatic nodules and bile duct epithelia of cirrhotic liver. To determine whether EGF mRNA accumulation accompanied a comparable increase in the EGF peptide, we performed immunohistochemistry using an antibody specific for the nonprocessed peptide aminoterminus. We observed that positive localized EGF staining paralleled its mRNA transcript. These results indicate that EGF upregulation is a characteristic of cirrhotic liver disease and suggest that persistent de novo ligand synthesis and its signaling contribute to an autocrine-mediated hepatocyte proliferation within the regenerative nodule. PMID- 10820156 TI - Applicability of different antibodies for immunohistochemical localization of CFTR in sweat glands from healthy controls and from patients with cystic fibrosis. AB - The hereditary disease cystic fibrosis (CF) is caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Understanding of the consequences of CFTR gene mutations is derived chiefly from in vitro studies on heterologous cell cultures and on cells hyperexpressing CFTR. Data from ex vivo studies on human tissue are scarce and contradictory, a fact which is in part explained by secondary tissue destruction in most affected organs. The purpose of this study was to establish conditions under which wild-type and mutated CFTR can be studied in affected human tissue. Sweat glands carry the basic defect underlying CF and are not affected by tissue destruction and inflammation. Therefore, we used this tissue to test a panel of eight different CFTR antibodies under various fixation techniques. The antibodies were tested on skin biopsy sections from healthy controls, from CF patients homozygous for the most common mutation, DeltaF508, and from patients carrying two nonsense mutations. Of the eight CFTR antibodies, only three-M3A7, MATG 1104, and cc24-met the criteria necessary for immunolocalization of CFTR in sweat glands. The labeling pattern in the CF sweat glands was consistent with the postulated processing defect of DeltaF508 CFTR. The antibodies exhibited different sensitivities for detecting DeltaF508 CFTR. PMID- 10820157 TI - Immunocytochemical evidence suggesting that diamine oxidase catalyzes biosynthesis of gamma-aminobutyric acid in antropyloric gastrin cells. AB - gamma-Aminobutyric acid (GABA) is a neurotransmitter that also occurs in a few non-neuronal cell types, where it may serve as a paracrine modulator. GABA is biosynthesized from glutamate by glutamate decarboxylase (GAD) and from putrescine via diamine oxidase (DAO). GAD is demonstrable in several GABA positive cell types but is undetectable in the GABA-containing gastrin cells and somatostatin cells of the antropyloric mucosa of the stomach. Using two antisera raised against synthetic peptides corresponding to two different regions of rat DAO, we now demonstrate strong reactivity for DAO in gastrin-positive cells of the rat antropyloric mucosa, whereas somatostatin-positive cells as well as other structures of the antrum are unreactive. Western blotting analysis of antrum and colon demonstrate that both antisera react with a single band of 85 kD, consistent with the predicted molecular weight of DAO. Expression of DAO mRNA in the antrum is demonstrated by reverse transcriptase polymerase chain reaction (RT PCR). Our results strongly indicate that gastrin cells produce GABA via DAO catalyzed oxidation of putrescine, and experimental data moreover suggest that the biosynthesis of GABA is regulated by the prandial state. Because GABA modulates release of somatostatin, these results point to a new mechanism of paracrine interaction between gastrin cells and somatostatin cells. PMID- 10820158 TI - Analysis of tissue chimerism in nude mouse brain and abdominal xenograft models of human glioblastoma multiforme: what does it tell us about the models and about glioblastoma biology and therapy? AB - In situ hybridization coupled to immunohistochemistry for antigens of interest allows unequivocal identification of tumor cells from reactive stroma cells and normal adjacent structures in human glioblastoma multiforme grafts transplanted into nude mice. With this methodology, we have explored the development of glioblastoma multiforme solid grafts transplanted into nude mouse brains or flanks. The brain transplants closely resembled the human situation, particularly in relation to differentiation and growth patterns. The morphological features of peritumoral reactive gliosis were similar to those observed in humans. A mouse glial stroma within the main tumor masses was also demonstrated. Kinetic studies showed that the compartment of isolated tumor cells that infiltrated host brains and the reactive gliosis constituted two cycling cell populations. Despite VEGF protein expression by tumor cells and some reactive astrocytes, the abnormally permeable microvascular beds were not hyperplastic. The observation of a non infiltrative pattern of growth when grafts were established in host flanks demonstrated that the organ-specific environment plays a determining role in the growth and invasive properties of glioblastoma. The phylogenetic distance between man and mouse and the recipient immunoincompetence should not impose serious limitations on the use of this model for studying malignant glioma biology and therapy in vivo. PMID- 10820159 TI - Luminal localization of blood-brain barrier sodium, potassium adenosine triphosphatase is dependent on fixation. AB - Cytochemical data in the literature reporting localization of sodium, potassium adenosine triphosphatase (Na(+), K(+)-ATPase) in the blood-brain barrier (BBB) have been contradictory. Whereas some studies showed the enzyme to be located exclusively on the abluminal endothelial plasma membrane, others demonstrated it on both the luminal and abluminal membranes. The influence of fixation on localization of the enzyme was not considered a critical factor, but our preliminary studies showed data to the contrary. We therefore quantitatively investigated the effect of commonly used fixatives on the localization pattern of the enzyme in adult rat cerebral microvessels. Fixation with 1%, 2%, and 4% formaldehyde allowed deposition of reaction product on both the luminal and abluminal plasma membranes. The luminal reaction was reduced with increasing concentration of formaldehyde. Glutaraldehyde at 0.1%, 0.25%, 0.5%, in combination with 2% formaldehyde, drastically inhibited the luminal reaction. The abluminal reaction was not significantly altered in all groups. These results show that luminal localization of BBB Na(+), K(+)-ATPase is strongly dependent on fixation. The lack of luminal localization, as reported in the literature, may have been the result of fixation. The currently accepted abluminal polarity of the enzyme should be viewed with caution. PMID- 10820160 TI - Distribution of NADPH-diaphorase and nitric oxide synthase (NOS) in different regions of porcine oviduct during the estrous cycle. AB - Nitric oxide synthase (NOS) is responsible for the biological production of nitric oxide (NO) in several organs, including those of the reproductive tract. We investigated potential changes in NADPH-diaphorase (NADPH-d) activity (marker for NOS activity) and the presence and distribution of NOS in the porcine oviduct. Tissues were obtained from gilts (n=16) on different days of the estrous cycle. One fallopian tube was used for histo- and immunohistochemistry and the other for Western blotting analysis. NADPH-d activity was much higher in the epithelium of the mucosa than in the myosalpinx. The highest activity of NADPH-d was always found in the epithelium of the isthmus. The intensity of the reaction (arbitrary units +/- SEM) in isthmus epithelium increased from the postovulatory period until early proestrus (96.2 +/- 11.2) and then gradually decreased. The lowest intensity of NADPH-d reaction in the epithelium of the isthmus was seen at estrus (58.4 +/- 7.7). The most intense NADPH-d activity in myosalpinx of all parts of the oviduct was observed at the postovulatory stage of the estrous cycle (isthmus 38.3 +/- 2.5; ampulla 35.6 +/- 4.2; infundibulum 24.7 +/- 0.8) and then decreased during the remaining stages of the estrous cycle (p< 0.001). The presence of endothelial NOS (eNOS) was detected in epithelial cells of mucosa and in endothelium of vascular tissues and myosalpinx during all studied days of the estrous cycle. The positive reaction for inducible NOS (iNOS) was restricted only to the endothelium of lymph vessels and some blood vessels. Because our Western blotting analysis revealed that porcine oviduct contains eNOS but not iNOS, we suggest that eNOS is the main isoform of NOS expressed in the porcine oviduct. We concluded that the different activity of NADPH-d in the various regions of the oviduct, accompanied by changes in its activity during the course of the estrous cycle, could indicate an important role of NO in regulation of tubal function. PMID- 10820161 TI - A new method to visualize the Helicobacter pylori-associated Lewis(b)-binding adhesin utilizing SDS-digested freeze-fracture replica labeling. AB - Freeze-fracture replica labeling has become a versatile tool to visualize both membrane components and other cell structures using SDS-replica cleaning before specific immunogold labeling of proteins or lipids. We report here for the first time the adoption and optimization of the method to studies of bacterial envelopes, as applied to structural analysis of the distribution of the unique BabA-adhesin of the gastric pathogen Helicobacter pylori. BabA is important for bacterial adherence to the human epithelial cell lining of the stomach. The adhesin was found to be distributed all over the bacterial cell surfaces. Our results suggest that the SDS-replica labeling allows assessment of protein localization to distinct cell compartments and analysis of co-localization with neighboring membrane structures. PMID- 10820162 TI - Primary structure of mouse and rat nephrin cDNA and structure and expression of the mouse gene. AB - Nephrin is a central component of the glomerular podocyte slit diaphragm and is essential for the normal renal filtration process. This study describes the complete structure of the mouse nephrin gene, which was shown to be homologous to the human gene, the major difference being 30 exons in the mouse gene as opposed to 29 in human. The complete primary structure of mouse and rat nephrins was also determined. The sequence identity between the mouse and rat proteins was shown to be 93%, while both rodent proteins have only about 83% sequence identity with human nephrin. The availability of the three mammalian sequences is significant for the interpretation of sequence variants and mutations in the nephrin gene in patients with congenital nephrotic syndrome. In situ hybridization analyses of whole mouse embryos and tissues revealed high expression of nephrin in kidney glomeruli and, surprisingly, an intense and highly restricted expression in a set of cells in hindbrain and spinal cord. No expression was observed elsewhere. This expression pattern may explain occasionally occurring neural symptoms caused by inactivating mutations in the nephrin gene in patients with congenital nephrotic syndrome. PMID- 10820163 TI - Effect of serotonin receptor antagonist on phosphate excretion. AB - To determine whether endogenous intrarenal 5-hydroxytryptamine affects phosphate excretion, the serotonin receptor antagonist methiothepin (20 microgram/kg, +6 microgram/kg per h) was infused into the renal interstitium of rats fed a normal phosphate diet (0.7% phosphate [Pi]) in the presence of endogenous parathyroid hormone (PTH). Renal interstitial infusion of methiothepin significantly increased fractional phosphate excretion (FE(Pi)) from 23 +/- 4 to 30 +/- 4% (n = 8, P < 0.05). To determine whether serotonin modulates the phosphaturic response to PTH during conditions of dietary phosphate excess or deprivation, rats were fed either a high (1.8% Pi, HPD) or low (0.07% Pi, LPD) phosphate diet, and methiothepin (100 microgram/kg, +30 microgram/kg per h) or saline vehicle was infused intravenously before and during PTH infusion (33 U/kg, +1 U/kg per min). Methiothepin infusion significantly increased FE(Pi) in thyroparathyroidectomized rats fed a HPD from 25 +/- 4 to 32 +/- 4% (n = 9, P < 0.05), and the subsequent administration of PTH further increased the FE(Pi) to 64 +/- 3% (P < 0.05). The increase in FE(Pi) during PTH infusion was similar in the absence (Delta27 +/- 5%, n = 7) and presence (Delta33 +/- 6%) of methiothepin, P > 0.05. In thyroparathyroidectomized rats fed a LPD, methiothepin infusion did not increase phosphate excretion (0.8 +/- 0.4 to 1.3 +/- 0.9%, n = 7, P > 0.05). However, the increase in FE(Pi) during PTH infusion was significantly greater in the presence of methiothepin (1.3 +/- 0.9 to 20.0 +/- 4.0%, Delta18.7 +/- 3.5%) than in the vehicle-infused rats (0.5 +/- 0.2 to 8.8 +/- 1.1%, Delta8.3 +/- 1.2%; n = 8, P < 0.05). In conclusion, these observations suggest that endogenous intrarenal serotonin enhances phosphate reabsorption in phosphate-replete rats, and attenuates the phosphaturic response to PTH in phosphate-deprived rats. PMID- 10820164 TI - Luminal and basolateral membrane transport of glutathione in isolated perfused S(1), S(2), and S(3) segments of the rabbit proximal tubule. AB - Lumen-to-bath and bath-to-lumen transport rates of glutathione (GSH) were measured in isolated perfused S(1), S(2), and S(3) segments of the rabbit proximal tubule. In lumen-to-bath experiments, the perfusion solution contained 4.6 microM (3)H-GSH with or without 1.0 mM acivicin. In all three segments perfused without acivicin, luminal disappearance rate (J(DL)) and bath appearance rate (J(AB)) of (3)H-GSH were 14.5 +/- 0.5 and 2.2 +/- 0.8 fmol/min per mm tubule length, respectively. With acivicin present, J(DL) and J(AB) were reduced to 1.3 +/- 0.4 and 0.5 +/- 0.3, respectively, with no differences among segments. Cellular concentrations of (3)H-GSH in S(1), S(2), and S(3) segments when acivicin was absent were 23.1 +/- 2.0, 31.7 +/- 11.4, and 143.5 +/- 17.9 microM, respectively. With acivicin in perfusate, cellular concentrations were reduced but there was no change in the heterogeneity profile. In bath-to-lumen transport experiments (S(2) segments only), the bathing solution contained 2.3 microM (3)H GSH. (3)H-GSH appearance in the lumen (J(AL), fmol/min per mm) and cellular accumulation from the bath were studied with and without acivicin in the perfusate. J(AL) values were 3.0 +/- 0.2 and 0.2 +/- 0.03 while cellular concentrations were 9.5 +/- 1.0 and 6.1 +/- 0.5 microM, respectively. It is concluded that: (1) GSH is primarily removed from the luminal fluid after degradation to glycine, cysteine, and glutamate, which are absorbed; (2) GSH can be absorbed intact at the luminal membrane; (3) the S(3) segment has the greatest GSH cellular concentration because its basolateral membrane has less capacity for cell-to-bath transport of GSH; and (4) GSH can be secreted intact from the peritubular compartment into the tubular lumen. PMID- 10820165 TI - Regenerative and proinflammatory effects of thrombin on human proximal tubular cells. AB - Interstitial fibrin deposition is a common histologic feature of tubulointerstitial diseases, which suggests that the coagulation system is activated. Thrombin, generated during the activation of the coagulation cascade, is a powerful activating factor for different cell types. Although proximal tubular cells are potential targets for this coagulation factor, no information is available on the effect of thrombin on these cells. Thus, the expression of protease-activated receptor-1 (PAR-1), the main thrombin receptor, was investigated in human proximal tubular cells (hPTC) in vivo and in vitro. A diffuse expression of PAR-1 was observed by immunohistochemistry along the basolateral membrane of PTC in normal human kidney. This observation was confirmed in vitro in cultured hPTC. Because tubular damage and monocyte infiltration are two hallmarks of tubulointerstitial injury, the effect of thrombin on DNA synthesis and monocyte chemotactic peptide-1 (MCP-1) gene and protein expression was evaluated in cultured hPTC. Thrombin induced a significant and dose-dependent increase in thymidine uptake and a striking upregulation of MCP-1 mRNA expression and protein release into the supernatant. Although PAR-1 is a G protein-coupled receptor, its activation in hPTC, as in other cell systems, resulted in a transient increase in cellular levels of tyrosine-phosphorylated proteins. An increased level of tyrosine-phosphorylated c-src suggested the activation of this cytoplasmic tyrosine kinase in response to thrombin and its potential role in thrombin-induced protein-tyrosine phosphorylation. Interestingly, thrombin-induced DNA synthesis and MCP-1 gene expression were completely blocked by genistein, a specific tyrosine kinase inhibitor, but not by its inactive analogue daidzein, demonstrating a central role for tyrosine kinase activation in the thrombin effects on hPTC. Moreover, the specific src inhibitor PP1 abolished the thrombin effect on DNA synthesis. In conclusion, thrombin might represent a powerful regenerative and proinflammatory stimulus for hPTC in acute and chronic tubulointerstitial diseases. PMID- 10820166 TI - Mitogen-activated protein kinase phosphorylation in kidneys of beta(s) sickle cell mice. AB - Previous studies in beta(s) sickle cell mice demonstrated renal immunostaining for nitrotyrosine, which is putative evidence of peroxynitrite (ONOO(-)) formation. ONOO(-) is known to nitrate tyrosine residues of various enzymes, thereby interfering with phosphorylation and inactivating them. The present study examined the state of phosphorylation of mitogen-activated protein (MAP) kinase signal transduction enzymes, i.e., p38, c-Jun NH(2)-terminal kinase (JNK), and extracellular signal-regulated kinase (ERK). Western blot performed with antibodies directed against specific phosphorylated threonine/tyrosine residues of these enzymes demonstrated reduced phosphorylation of renal p38 and a trend toward reduced phosphorylation of ERK. In contrast, phosphorylation of renal JNK was markedly increased compared with normal mice. The abundance of MAP kinase phosphatase-1 (MKP-1), a key upstream enzyme that modulates phosphorylation of MAP kinases, was not different in beta(s) versus normal mice. To determine whether nitration of tyrosine by ONOO(-) was responsible for reduced phosphorylation of p38 and ERK, mercaptoethylguanidine (MEG), a compound known to reduce inducible isoform of nitric oxide synthase activity and to scavenge ONOO( ), was administered to beta(s) mice for 5 d. MEG was found to restore phosphorylation of p38 and ERK toward normal levels. These observations provide evidence that ONOO(-) (or closely related reaction products of NO) contributes to dephosphorylation of p38 and ERK, and presumably reduces activity of these enzymes. The increased phosphorylation of JNK, which suggests activation of this signaling pathway by extracellular stress signals, may play a role in apoptosis in the kidneys of these mice. The changes in phosphorylation of MAP kinase pathways found in this study could have important consequences for regulation of nuclear transcription factors, and thus renal function and pathology in sickle cell kidneys. PMID- 10820167 TI - Nephrogenic diabetes insipidus: functional analysis of new AVPR2 mutations identified in Italian families. AB - The aim of this study was to identify loss-of-function mutations of the V2 vasopressin receptor gene (AVPR2) in Italian patients affected by X-linked nephrogenic diabetes insipidus (NDI). Mutations were found in 15 of the 18 unrelated families investigated: nine of these mutations were previously unknown, including two affecting residues located in regions known to be important for determining the pharmacologic properties of the receptor, which were therefore functionally investigated. The first (A84D) involves a residue located near an aspartic acid (D85) that is highly conserved in all G protein-coupled receptors and that is believed to play a role in the process of their isomerization into functionally active and inactive states. The present study indicates that this mutation not only affects receptor folding in such a way as to lead to its retention inside the intracellular compartments but, as expected, also has profound effects on its binding and coupling properties. The second was a mutation of a tryptophan located at the beginning of the first extracellular loop (W99R) that greatly impaired the binding properties of the receptor and had a minor effect on its intracellular routing. Molecular analysis of the first extracellular loop bearing this mutation suggests that this residue plays a fundamental role in stabilizing the peptide/receptor interactions responsible for the high-affinity binding of agonists to the V2 receptor. PMID- 10820168 TI - Report of 33 novel AVPR2 mutations and analysis of 117 families with X-linked nephrogenic diabetes insipidus. AB - X-linked nephrogenic diabetes insipidus (NDI) is a rare disease caused by mutations in the arginine vasopressin receptor 2 gene (AVPR2). Thirty-three novel AVPR2 mutations were identified in 62 families that were not included in our previous studies. This study describes the diversity of mutations observed in a total of 117 families, the number of affected people at the time of diagnosis, skewed X chromosome inactivation in severely affected females, the inferred parental origin of de novo mutations, and it provides estimates of incidence. Among 117 families, there were 82 different putative disease-causing mutations. Based on haplotype analysis, it can be inferred that when the same AVPR2 mutation is identified in different families that were not known to be related, the mutations most likely arose independently. More than half of the families had only one affected male; two families presented with a severely affected female and no family history of NDI. A de novo mutation arose during oogenesis in the mother in 20% of isolated cases. The estimate of about 8.8 per million male live births of the incidence of X-linked NDI in the province of Quebec, Canada may be representative of the general population except in Nova Scotia and New Brunswick, where the incidence is more than six times higher. Documentation of the diversity of mutations will assist in revealing the full spectrum of clinical variation. Discussion of genetic and population genetic aspects of X-linked NDI may contribute to early diagnosis and treatment. PMID- 10820169 TI - Angiopoietin-2 is a site-specific factor in differentiation of mouse renal vasculature. AB - Angiopoietin-1 (Ang-1) stimulates endothelial and vascular network differentiation through the Tie-2 receptor tyrosine kinase, while Ang-2 modulates this activation in embryo and tumor growth. The nephrogenic pattern of Ang-2 was documented in a mouse strain that expresses the LacZ reporter gene driven by the Ang-2 promoter. Heterozygous animals were healthy with morphologically normal kidneys, and they were examined after X-gal staining. At embryonic days 10.5 (E10.5) and E12.0, transgene expression was absent in the mesonephros and metanephros. At E14.0, expression was noted in the metanephric artery and its major branches. At E19.0 and in neonatal kidneys, expression was maintained in larger renal artery branches, extending to arcuate and smaller cortical vessels. Histologically, transgene expression was located in multiple layers of vessel wall cells, extending further from the endothelium than alpha-smooth muscle actin. The mesangium of immature glomeruli also expressed LacZ. In the first 3 postnatal weeks, a new pattern became evident, with intense X-gal staining in the inner stripe of the outer medulla, where a subset of thin descending limbs of loops of Henle expressed the transgene. This dynamic and developmentally regulated pattern indicates that Ang-2 is an early marker of the renal pericyte and vascular smooth muscle lineage and is also an epithelial-derived growth factor. Because Tie-2 is widely expressed by differentiating renal endothelia, this study is consistent with the hypothesis that Ang-2 has roles in kidney vascular maturation. PMID- 10820170 TI - Course of renal injury in the Mpv17-deficient transgenic mouse. AB - The mutant Mpv17 mouse is a transgenic strain that fails to express a protein that is normally expressed in the kidney and that is associated with peroxisomes. The present studies provide a quantitative examination of renal function and structure in this strain compared to its control CFW strain. By 52 wk of age, the mutant strain developed proteinuria (urinary protein to creatinine ratio: 25 +/- 14 versus 3 +/- 1, mutant versus control), albuminuria (urinary albumin to creatinine ratio: 23 +/- 15 versus 0.1 +/- 0.1, mutant versus control), and hypoalbuminemia (2.1 +/- 0.4 versus 2.5 +/- 0.2 G/dl, mutant versus control), but without arterial hypertension or major reduction in filtration (serum creatinine 0.14 +/- 0.04 versus 0.18 +/- 0.12 mg/dl, mutant versus control). The Mpv17 glomeruli were enlarged (0.98 +/- 0.12 versus 0.52 +/- 0.02 micrometer(3) x 10(6), mutant versus control). Glomerular sclerosis became widespread (95 +/- 3 versus 23 +/- 32%, mutant versus control) and was preceded by mesangiolysis and microaneurysms. Tubulointerstitial disease was conspicuous by its absence. The intrarenal vasculature was normal in the mutant mice. Electron microscopy demonstrated focal foot process fusion and mesangiolysis. Thus, this mutant strain of mouse develops proteinuria and a distinct glomerulopathy including mesangiolysis but little interstitial injury all due to the loss of expression of a single gene. PMID- 10820171 TI - Functional relevance of activated beta1 integrins in mercury-induced nephritis. AB - Cell adhesion through different adhesion molecules is a crucial event in the inflammatory response. Integrins can only bind and mediate cellular adhesion after their activation by different specific stimuli. The state of beta1 integrin activation can be assessed by a group of monoclonal antibodies (HUTS) that selectively recognize beta1 integrins in their active form. A similar activated epitope in the rat was defined using the anti-human monoclonal antibody HUTS-21, which recognizes an activation-dependent epitope on the beta1 chain. It was found that the divalent cations Mn(2+) and Hg(2+) were able to induce in vitro the activation of beta1 integrins on rat lymphocytes. The Hg(2+) cation induces an autoimmune disease in the Brown Norway rat characterized by synthesis and glomerular deposits of anti-glomerular basement membrane antibodies, proteinuria, and interstitial nephritis. Using the mercury model of nephritis, it was found that the expression of HUTS-21 epitope is induced in vivo in rat lymphocytes, and its appearance is correlated with the other parameters at the onset of the disease. In addition, the administration of HUTS-21 monoclonal antibody to HgCl(2)-treated rats offered evidence of its protective effects (1) against infiltration of renal interstitium by leukocytes, and (2) in the reduction of anti-glomerular basement membrane synthesis and glomerular deposition. Nevertheless, urinary protein values remained unaffected. These results demonstrate a key role of beta1-activated integrins in both leukocyte cell-cell interactions and leukocyte infiltration pathway mechanism, and also indicate that leukocyte migration may have less importance in the development of this disease than previously thought. PMID- 10820172 TI - C-terminal parathyroid hormone-related protein increases vascular endothelial growth factor in human osteoblastic cells. AB - The N-terminal region of parathyroid hormone (PTH) and PTH-related protein (PTHrP) interacts with a common PTH/PTHrP receptor in osteoblasts. These cells synthesize PTHrP, but its role in bone turnover is unclear. Intermittent treatment with N-terminal PTHrP or PTH stimulates bone growth in vivo, possibly by increasing local bone factors. In addition, C-terminal PTHrP (107-139), which does not bind to the PTH/PTHrP receptor, appears to affect bone resorption in vivo and in vitro, although its effect on bone formation in vivo remains controversial. Bone angiogenesis is an often overlooked but critical event in the process of bone remodeling. Recently, PTH (1-34) has been shown to induce gene expression of vascular endothelial growth factor (VEGF), a potent angiogenic factor, by osteoblastic cells. However, no data are available on the effect of PTHrP (107-139) on VEGF expression in these cells. Using semiquantitative reverse transcription followed by PCR, we found that PTHrP (107-139), between 10 nM and 1 pM, increased VEGF mRNA in human osteoblastic (hOB) cells from trabecular bone. This effect of this agonist, at 10 nM, was maximal (fivefold for VEGF(165), and twofold for VEGF(121), compared to control) within 1 to 4 h. This effect was similar to that induced by PTHrP (1-34) in these cells, as well as in human osteosarcoma MG-63 cells, using Northern blot analysis. Moreover, the effect of both peptides, added together at 100 pM, was not higher than that observed with each peptide alone in hOB cells. The effects of PTHrP (107-139) and that of PTHrP (1-34) were abolished by actinomycin D in hOB cells. In these cells, the protein kinase C inhibitor staurosporine, but not the protein kinase A inhibitor H89, inhibited the increase in VEGF mRNA induced by 10 nM PTHrP (107-139). PTHrP (107 139), at 10 nM, also stimulated cytosolic VEGF immunostaining in hOB cells, and VEGF secretion into the medium conditioned by hOB or MG-63 cells for 24 h, which was (ng/mg protein): 10 +/- 1 or 5 +/- 3 (control), respectively, and 21 +/- 1 or 11 +/- 2 (PTHrP [107-139]-stimulated), respectively. Furthermore, medium conditioned by these cells for 24 h in the presence of 10 nM PTHrP (107-139), with or without 10 nM PTHrP (1-34), increased about 30% bovine aortic endothelial cell (BAEC) growth at 48 h. This effect was inhibited by adding a specific anti VEGF antibody to the BAEC incubation medium. These findings demonstrate that the C-terminal domain of PTHrP induces expression and secretion of VEGF, a main angiogenic factor, in hOB cells and MG-63 cells. This relationship between PTHrP and VEGF has potential implications for both bone vascularization and bone formation, and neoangiogenesis in PTHrP-producing tumors. PMID- 10820173 TI - High prevalence of low bone turnover and occurrence of osteomalacia after kidney transplantation. AB - Kidney transplantation corrects most of the metabolic abnormalities that cause renal osteodystrophy. However, many transplanted patients develop osteoporosis and other bone lesions that are related, at least in part, to their immunosuppressive regimen. The precise histologic patterns of bone disease after transplantation are not well defined. In a study designed to investigate this issue, 57 adult posttransplant patients agreed to undergo bone biopsies and blood drawings. There were 32 men and 25 women, mean age 45 +/- 2 yr, who had received a kidney transplantation 5.6 +/- 0.8 yr before biopsy. History of bone pain, fractures, and avascular necrosis was found in 22, 12, and 7 patients, respectively. Serum creatinine was 1.68 +/- 0.1 mg/dl, 21% of patients were hypercalcemic, 63.2% had elevated parathyroid hormone (PTH) (>65 pg/ml), and 91.2% had normal calcitriol levels. Cancellous bone volume/tissue volume was below normal compared to age- and gender-matched control subjects in 56.1% of patients. Bone turnover (activation frequency) was low in 45.6%, normal in 28.1%, and elevated in 26.3% of patients. Bone formation rate/bone surface was low in 59.7%, normal in 35%, and elevated in 5. 3% of the patients. Erosion surface/bone surface was high in 21.1% of patients. Mineralization was prolonged in 87.5% of patients, including 9 patients with osteomalacia and 12 patients with focal osteomalacia. Cumulative and maintenance doses of prednisone and time elapsed since transplantation correlated negatively with bone volume and bone turnover (r = -0.32 to -0.59, P < 0.05 to 0.01), whereas cumulative doses of cyclosporine or azathioprine, age, gender, or serum PTH levels did not. Regression analysis identified prednisone as the main factor responsible for low bone volume and bone turnover (r = 0.54 and r = 0.43, P < 0.01). No factors were found to predict delayed mineralization. The present study shows that low bone volume, low bone turnover, and generalized or focal osteomalacia are frequent histologic features in transplanted patients. The effects of age, gender, PTH, and cyclosporine on bone volume and bone turnover are apparently overridden by the prominent effects of glucocorticoids. The prevalence of mineralization defect in the presence of normal serum levels of calcidiol and calcitriol suggests vitamin D resistance and deserves further study. PMID- 10820174 TI - Urinary protein binding does not affect response to furosemide in patients with nephrotic syndrome. AB - Response to loop diuretics in patients with nephrotic syndrome (NS) is subnormal. Studies in animal models of NS have suggested that binding of diuretic to urinary albumin is one of the mechanisms that may be operative in this diuretic resistance. To explore this hypothesis, 12 patients with NS were studied to determine whether displacement from urinary protein binding with sulfisoxazole would restore response to 120 mg of furosemide. The study was stopped after treating seven patients because it was clear that sulfizoxazole had no effect. Sodium excretion (mean +/- SD) from furosemide alone was 239 +/- 90 versus 240 +/ 115 mEq/8 h with sulfisoxazole. Sulfisoxazole had modest effects on serum pharmacokinetics of furosemide but had no effect on either the time course of furosemide urinary excretion or overall amount excreted: 49 +/- 15 mg versus 54 +/- 12 mg for furosemide alone and furosemide plus sulfisoxazole, respectively. It is concluded that urinary protein binding of loop diuretics is not a major mechanism for the diuretic resistance of NS. In turn, strategies aimed at displacing such binding are unlikely to be clinically helpful. PMID- 10820175 TI - Effect of high dose folic acid therapy on hyperhomocysteinemia in hemodialysis patients: results of the Vienna multicenter study. AB - Homocysteine is associated with atherosclerosis and enhanced cardiovascular risk. In previous studies, treatment with folic acid up to 15 mg/d failed to correct hyperhomocysteinemia in the majority of end-stage renal disease patients. A dose of 30 or 60 mg of folic acid per day was compared with 15 mg/d in an attempt to normalize hyperhomocysteinemia in 150 hemodialysis patients. In a randomized, double-blind, multicenter study, 144 patients completed the 4-wk treatment period and 121 patients completed the 6-mo follow-up. Total homocysteine plasma levels were reduced by 32.1% (15 mg/d), 29. 9% (30 mg/d), or 37.8% (60 mg/d) with no significant differences found between the three treatment groups. Baseline total homocysteine plasma concentration was an independent predictor of the response to folic acid therapy (P = 0.0001), whereas the 5, 10-methylenetetrahydrofolate reductase polymorphisms (MTHFR 677C --> T and 1298A --> C) had no influence. Nevertheless, patients with the MTHFR 677TT genotype more frequently attained normal total homocysteine plasma levels than patients with the CC or CT genotype (P = 0.025). In response to 60 mg of folic acid per day, TT genotype patients had lower folate plasma levels compared to CC or CT genotype patients (P = 0.016). After completion of the 4-wk treatment period with 30 or 60 mg of folic acid per day, there was a marked rebound of total homocysteine plasma levels at the end of the follow-up in patients with the MTHFR 677TT genotype, which even exceeded baseline values in several patients (P = 0.0001). This study clearly demonstrates that doses of 30 or 60 mg of folic acid per day are not more effective than 15 mg/d in reducing hyperhomocysteinemia in regular hemodialysis patients. Patients with the MTHFR 677TT genotype are more likely to realize normal total homocysteine plasma levels. Folic acid at 30 or 60 mg/d but not 15 mg/d results in a rebound of total homocysteine plasma concentrations when treatment is stopped. PMID- 10820176 TI - Pharmacokinetics of once daily intraperitoneal cefazolin in continuous ambulatory peritoneal dialysis patients. AB - This study determined the pharmacokinetic characteristics of once daily intraperitoneal (IP) cefazolin in continuous ambulatory peritoneal dialysis (CAPD) patients. Each of the 10 volunteer CAPD patients without active peritonitis received a single IP dose of 1 g of cefazolin sodium for a 6-h dwell. All patients underwent a fixed CAPD regimen comprising a first 6-h dwell followed by two 3-h dwells and a final 12-h overnight dwell. Blood and dialysate samples were collected at 0, 0.5, 1, 2, 3, 6 (end of first dwell), and 24 h after the administration of IP cefazolin. Any urine produced was collected over the 24-h study period. A validated HPLC method was used to analyze cefazolin in plasma, dialysate, and urine. The bioavailability was found to be 77.9 +/- 3.1%, volume of distribution 0.20 +/- 0.05 L/kg, and plasma half-life 39.9 +/- 25.4 h. Mean total, renal, and peritoneal clearances were 4.5 +/- 2.3, 1. 4 +/- 1.1, and 3.5 +/- 1.8 ml/min, respectively. Mean plasma and dialysate concentrations at 24 h were 42.8 +/- 14.3 and 31.8 +/- 11. 7 mcg/ml, respectively, well above the minimum inhibitory concentrations (MIC) of susceptible organisms. A once daily IP cefazolin dose of 500 mg/L gave desirable pharmacokinetic attributes for use as a suitable alternative to vancomycin for empiric treatment of CAPD-associated peritonitis. PMID- 10820177 TI - Low intraindividual variability of cyclosporin A exposure reduces chronic rejection incidence and health care costs. AB - The present study applied a receiver operating characteristic (ROC) analysis to assess the role of intraindividual variability of cyclosporin A (CsA) drug exposure in predisposing renal transplant recipients to the occurrence of chronic rejection, as well as to increased health care costs using a resource-based economic analysis. Two hundred and four adult renal transplant recipients were treated with tapering doses of prednisone (Pred) and with a concentration controlled strategy that selected doses of the olive oil-based formulations of CsA (Sandimmune(R)) that achieved target concentrations based on serial pharmacokinetic profiles. The ROC analysis revealed an inflection point of plots of the coefficient of variation (%CV) of CsA exposure versus the risk of chronic rejection at >/=28.4% for the average concentration (C(av)), i.e., the dosing interval-corrected area under the concentration-time curves, and >/=36% for the trough concentration (C(0)). The incidence of chronic rejection over a period of 5 yr was 24% among the less variable (LV) versus 40% among the variable (V) cohort. The economic analysis revealed that the total mean facility and physician costs per patient were $48,789 versus $60,998, respectively (P < 0.01). The degree of variability displayed by any individual could only be predicted by serial measurements of CsA concentrations, and not by demographic features, laboratory determinations, clinical characteristics, individual or mean values of any observed CsA concentration, or other pharmacokinetic parameters calculated following a single drug exposure. Thus, strategies that reduce intrapatient variability of CsA exposure over time may lead to reductions in chronic allograft loss and in treatment costs. PMID- 10820178 TI - Thrombotic microangiopathy associated with parvovirus B 19 infection after renal transplantation. AB - Human parvovirus B19 is considered an etiologic agent of aplastic anemia in immunosuppressed patients. Microscopic vasculitis, with or without renal involvement, has recently been attributed to this viral infection in immunocompetent patients. This study describes four cases of thrombotic renal graft microangiopathy presumably secondary to B19 infection. Twelve to 50 days after transplantation, four patients presented a renal graft dysfunction with creatinine rising to 360 to 1088 micromol/L and requiring hemodialysis in three cases. Renal involvement appeared after a systemic illness characterized by fever, fatigue and arthralgia, aplastic anemia (hemoglobin ranged from 5.3 to 7.8 g/dl), and thrombocytopenia. A thrombotic microangiopathy was observed in the renal biopsies, and the parvovirus B19 genome was isolated by PCR from the specimens. All four patients also became IgM-positive for parvovirus. Three of the four renal biopsies taken at the time of transplantation (T0) from the same patients were found positive for the B19 genome. Graft function recovered, with resolution of the aplastic anemia, within 22 to 110 d. Twenty biopsies performed as routine controls or for suspected acute rejection and nine T0 biopsies of patients with no signs of B19 infection were used. The B19 genome was found in two of 20 posttransplant biopsies and in one of nine T0 biopsies. The temporal association between aplastic anemia and the onset of thrombotic graft microangiopathy, isolation of the viral genome in renal specimens, seroconversion, and endothelial tropism of the virus suggests that B19 could be the etiologic agent of thrombotic microangiopathy in these cases. The development of the disease after infection could depend on other detrimental cofactors, which make the patient more susceptible to microthrombi formation in the renal microvasculature. The renal graft could represent the route of B19 transmission. PMID- 10820179 TI - Encrusted pyelitis of native kidneys. AB - This study reports the first four cases of encrusted pyelitis involving native kidneys. The clinical features, management, and outcome of these patients were analyzed. Predisposing factors were underlying urologic disease and/or urologic manipulations, debilitating diseases, hospitalization, and prolonged antibiotic therapies. Presenting symptoms were renal failure in three patients with ureteroileal urinary diversion and manifestations of cystitis in one patient. Computed tomography scan of the urinary tract was critical for diagnosis. Presence of struvite was demonstrated by crystalluria and infrared spectrophotometry analysis of the encrusted material. Corynebacterium urealyticum urinary infection was identified in one case. Surgery (one patient) and palliative ureteral diversion (one patient), respectively, led to death and end stage renal failure. Successful dissolution of encrusted pyelitis was obtained in two patients treated with intravenous vancomycin and local acidification of the renal collecting system. Clinical observation shows that encrusted pyelitis is a threatening disorder that destroys the native kidneys and may lead to end-stage renal failure. Successful treatment of the disease by chemolysis and antibiotics depends on correct and early diagnosis. Diagnosis required recognition of the predisposing factors, computed tomography imaging of the urinary tract, crystalluria, and identification of urea-splitting bacteria with prolonged culture on selective medium. PMID- 10820180 TI - Cell biology of parathyroid gland hyperplasia in chronic renal failure. PMID- 10820181 TI - The diabetic foot in the dialyzed patient. PMID- 10820182 TI - The salt-inducible kinase, SIK, is induced by depolarization in brain. AB - Membrane depolarization of neurons is thought to lead to changes in gene expression that modulate neuronal plasticity. We used representational difference analysis to identify a group of cDNAs that are induced by membrane depolarization or by forskolin, but not by neurotrophins or growth factors, in PC12 pheochromocytoma cells. One of these genes, SIK (salt-inducible kinase), is a member of the sucrose-nonfermenting 1 protein kinase/AMP-activated protein kinase protein kinase family that was also recently identified from the adrenal gland of rats treated with high-salt diets. SIK mRNA is induced up to eightfold in specific regions of the hippocampus and cortex in rats, following systemic kainic acid administration and seizure induction. PMID- 10820183 TI - Identification and cataloging of genes induced by long-lasting long-term potentiation in awake rats. AB - Maintenance of long-term potentiation (LTP) requires de novo gene expression. Here we report the direct isolation, using PCR-differential display, of genes whose expression level was altered after induction of long-lasting LTP in the hippocampus of freely moving awake rats. Differential display using 480 primer combinations revealed 17 cDNA bands that showed a reproducible change in expression level. These cDNAs represented at least 10 different genes (termed RM1 10), all of which showed up-regulation at 75 min after LTP induction and a return to basal expression levels within 24 h. Three of these genes were known only from expressed sequence tags (RM1-3), two were known genes whose up-regulation by LTP has not been described (GADD153/CHOP and ler5), and five were known genes whose up-regulation by LTP has already been reported (MAPK phosphatase, NGFI-A/zif268, vesl-1S/homer-1a, Ag2, and krox-20). We characterized the expression profiles of genes in the two former categories with respect to NMDA receptor dependency, tissue specificity, and developmental regulation using northern blotting and semiquantitative RT-PCR. The up-regulation of all five of these genes was NMDA receptor-dependent and correlated with the persistence of LTP, suggesting that these genes may play functional roles in prolonged LTP maintenance. PMID- 10820184 TI - Molecular mechanisms underlying the activity-linked alterations in acetylcholinesterase mRNAs in developing versus adult rat skeletal muscles. AB - The molecular mechanisms underlying the activity-linked plasticity of acetylcholinesterase (AChE) mRNA levels in mammalian skeletal muscle have yet to be established. Here, we demonstrate that denervation of adult muscle induces a dramatic (up to 90%) and rapid (within 24 h) decrease in the abundance of AChE mRNAs. By contrast, denervation of 14-day-old rats leads to a significantly less pronounced reduction (50% of control) in the expression of AChE mRNAs. Assessment of the transcriptional activity of the AChE gene reveals that it remains essentially unchanged in adult denervated muscles, whereas it displays an approximately two- to three-fold increase (p < 0.05) in denervated muscles from 2 to 14-day-old rats. In addition, we observed a higher rate of degradation of in vitro transcribed AChE mRNAs upon incubation with protein extracts from denervated muscles. Finally, UV-crosslinking experiments reveal that denervation increases the abundance of RNA-protein interactions in the 3' untranslated region of AChE transcripts. Taken together, these data suggest that the abundance of AChE transcripts in mature muscles is controlled primarily via posttranscriptional regulatory mechanisms, whereas in neo- and postnatal muscles, both transcriptional and posttranscriptional regulation appears critical in dictating AChE mRNA levels. Accordingly, the activity-linked transcriptional regulation of the AChE gene appears to demonstrate a high level of plasticity during muscle development when maturation of the neuromuscular junctions is still occurring. PMID- 10820185 TI - Evolutionarily distinct classes of S27 ribosomal proteins with differential mRNA expression in rat hypothalamus. AB - Using an in situ hybridization screen for cDNA clones of brain region-specific mRNAs, we isolated a rat transcript that encodes a ribosomal protein S27. Searching GenBank DNA databases, we found two S27 protein isoforms. One isoform, encoded by multiple genes, is extant in archaea and eukarya, but not bacteria. The second isoform appears to be recently evolved because it has been identified only in mammals. Multiple transcripts encode each isoform and exhibit different tissue expression patterns throughout rat brain and periphery, with abundant expression in the hypothalamus. In situ hybridization studies revealed predominant expression of S27(1) in distinct hypothalamic nuclei, such as the paraventricular, supraoptic, suprachiasmatic, arcuate, and circularis nuclei, whereas expression of S27(2) mRNA was discretely expressed in select neurons of the periventricular and supraoptic nuclei. Combined with the genetic evidence that S27 has extraribosomal functions in plants, the complexity of S27 biology observed here may suggest auxiliary functions for S27 proteins in the mammalian nervous system. PMID- 10820186 TI - The Ginkgo biloba extract (EGb 761) protects and rescues hippocampal cells against nitric oxide-induced toxicity: involvement of its flavonoid constituents and protein kinase C. AB - An excess of the free radical nitric oxide (NO) is viewed as a deleterious factor involved in various CNS disorders. Numerous studies have shown that the Ginkgo biloba extract EGb 761 is a NO scavenger with neuroprotective properties. However, the mechanisms underlying its neuroprotective ability remain to be fully established. Thus, we investigated the effect of different constituents of EGb 761, i.e., flavonoids and terpenoids, against toxicity induced by NO generators on cells of the hippocampus, a brain area particularly susceptible to neurodegenerative damage. Exposure of rat primary mixed hippocampal cell cultures to either sodium nitroprusside (SNP; 100 microM) or 3-morpholinosydnonimine resulted in both a decrease in cell survival and an increase in free radical accumulation. These SNP-induced events were blocked by either EGb 761 (10-100 microg/ml) or its flavonoid fraction CP 205 (25 microg/ml), as well as by inhibitors of protein kinase C (PKC; chelerythrine) and L-type calcium channels (nitrendipine). In contrast, the terpenoid constituents of EGb 761, known as bilobalide and ginkgolide B, as well as inhibitors of phospholipases A [3-[(4 octadecyl)benzoyl]acrylic acid (OBAA)] and C (U-73122), failed to display any significant effects. Moreover, EGb 761 (50 microm) CP 205 (25 microg/ml), and chelerythrine were also able to rescue hippocampal cells preexposed to SNP (up to 1 mM). Finally, EGb 761 (100 microg/ml) was shown to block the activation of PKC induced by SNP (100 microM). These data suggest that the protective and rescuing abilities of EGb 761 are not only attributable to the antioxidant properties of its flavonoid constituents but also via their ability to inhibit NO-stimulated PKC activity. PMID- 10820187 TI - 4-Hydroxynonenal induces oxidative stress and death of cultured spinal cord neurons. AB - Primary spinal cord trauma can trigger a cascade of secondary processes leading to delayed and amplified injury to spinal cord neurons. Release of fatty acids, in particular arachidonic acid, from cell membranes is believed to contribute significantly to these events. Mechanisms of fatty acid-induced injury to spinal cord neurons may include lipid peroxidation. One of the major biologically active products of arachidonic acid peroxidation is 4-hydroxynonenal (HNE). The levels of HNE-protein conjugates in cultured spinal cord neurons increased in a dose dependent manner after a 24-h exposure to arachidonic acid. To study cellular effects of HNE, spinal cord neurons were treated with different doses of HNE, and cellular oxidative stress, intracellular calcium, and cell viability were determined. A 3-h exposure to 10 microM HNE caused approximately 80% increase in oxidative stress and 30% elevation of intracellular calcium. Exposure of spinal cord neurons to HNE caused a dramatic loss of cellular viability, indicated by a dose-dependent decrease in MTS [3-(4, 5-dimethylthiazol-2-yl)-5-(3 carboxymethoxyphenyl)-2-(4-s ulfophenyl)- 2H-tetrazolium, inner salt] conversion. The cytotoxic effect of HNE was diminished by pretreating neurons with ebselen or N-acetylcysteine. These data support the hypothesis that formation of HNE may be responsible, at least in part, for the cytotoxic effects of membrane-released arachidonic acid to spinal cord neurons. PMID- 10820188 TI - Expression of a dominant-negative mutant of p21(ras) inhibits induction of nitric oxide synthase and activation of nuclear factor-kappaB in primary astrocytes. AB - The present study underlines the importance of p21(ras) in regulating the inducible nitric oxide synthase (iNOS) in primary astrocytes. Bacterial lipopolysaccharides induced the GTP loading of p21(ras), and the expression of a dominant-negative mutant of p21(ras) (Deltap21(ras)) inhibited lipopolysaccharide induced GTP loading in rat primary astrocytes. To delineate the role of p21(ras) in the induction of iNOS, we examined the effect of Deltap21(ras) on the expression of iNOS and the production of nitric oxide. It is interesting that expression of Deltap21(ras) markedly inhibited the production of nitric oxide and the expression of iNOS in lipopolysaccharide- and proinflammatory cytokine (tumor necrosis factor-alpha, interleukin-1beta; interferon-gamma)-stimulated rat and human primary astrocytes. Inhibition of iNOS promoter-derived chloramphenicol acetyltransferase activity by Deltap21(ras) suggests that p21(ras) is involved in the transcription of iNOS. As activation of nuclear factor-kappaB (NF-kappaB) is necessary for the transcription of iNOS, we examined the effect of Deltap21(ras) on the activation of NF-kappaB. Expression of Deltap21(ras) inhibited the DNA binding as well as the transcriptional activity of NF-kappaB in activated astrocytes, suggesting that Deltap21(ras) inhibits the expression of iNOS by inhibiting the activation of NF-kappaB. These studies also suggest that inhibitors of p21(ras) may be used as therapeutics in nitric oxide- and cytokine mediated neuroinflammatory diseases. PMID- 10820189 TI - Dopamine-induced apoptosis is mediated by oxidative stress and Is enhanced by cyanide in differentiated PC12 cells. AB - Dopamine (DA) oxidation and the generation of reactive oxygen species (ROS) may contribute to the degeneration of dopaminergic neurons underlying various neurological conditions. The present study demonstrates that DA-induced cytotoxicity in differentiated PC12 cells is mediated by ROS and mitochondrial inhibition. Because cyanide induces parkinson-like symptoms and is an inhibitor of the antioxidant system and mitochondrial function, cells were treated with KCN to study DA toxicity in an impaired neuronal system. Differentiated PC12 cells were exposed to DA, KCN, or a combination of the two for 12-36 h. Lactate dehydrogenase (LDH) assays indicated that both DA (100-500 microM) and KCN (100 500 microM) induced a concentration- and time-dependent cell death and that their combination produced an increase in cytotoxicity. Apoptotic death, measured by Hoechst dye and TUNEL (terminal deoxynucleotidyltransferase dUTP nick end labeling) staining, was also concentration- and time-dependent for DA and KCN. DA plus KCN produced an increase in apoptosis, indicating that KCN, and thus an impaired system, enhances DA-induced apoptosis. To study the mechanism(s) of DA toxicity, cells were pretreated with a series of compounds and incubated with DA (300 microM) and/or KCN (100 microM) for 24 h. Nomifensine, a DA reuptake inhibitor, rescued nearly 60-70% of the cells from DA- and DA plus KCN-induced apoptosis, suggesting that DA toxicity is in part mediated intracellularly. Pretreatment with antioxidants attenuated DA- and KCN-induced apoptosis, indicating the involvement of oxidative species. Furthermore, buthionine sulfoximine, an inhibitor of glutathione synthesis, increased the apoptotic response, which was reversed when cells were pretreated with antioxidants. DA and DA plus KCN produced a significant increase in intracellular oxidant generation, supporting the involvement of oxidative stress in DA-induced apoptosis. The nitric oxide synthase inhibitor N(G)-nitro-L-arginine methyl ester and the peroxynitrite scavenger uric acid blocked apoptosis and oxidant production, indicating involvement of nitric oxide. These results suggest that DA neurotoxicity is enhanced under the conditions induced by cyanide and involves both ROS and nitric oxide-mediated oxidative stress as an initiator of apoptosis. PMID- 10820190 TI - Preferential resistance of dopaminergic neurons to the toxicity of glutathione depletion is independent of cellular glutathione peroxidase and is mediated by tetrahydrobiopterin. AB - Depletion of glutathione in the substantia nigra is one of the earliest changes observed in Parkinson's disease (PD) and could initiate dopaminergic neuronal degeneration. Nevertheless, experimental glutathione depletion does not result in preferential toxicity to dopaminergic neurons either in vivo or in vitro. Moreover, dopaminergic neurons in culture are preferentially resistant to the toxicity of glutathione depletion, possibly owing to differences in cellular glutathione peroxidase (GPx1) function. However, mesencephalic cultures from GPx1 knockout and wild-type mice were equally susceptible to the toxicity of glutathione depletion, indicating that glutathione also has GPx1-independent functions in neuronal survival. In addition, dopaminergic neurons were more resistant to the toxicity of both glutathione depletion and treatment with peroxides than nondopaminergic neurons regardless of their GPx1 status. To explain this enhanced antioxidant capacity, we hypothesized that tetrahydrobiopterin (BH(4)) may function as an antioxidant in dopaminergic neurons. In agreement, inhibition of BH(4) synthesis increased the susceptibility of dopaminergic neurons to the toxicity of glutathione depletion, whereas increasing BH(4) levels completely protected nondopaminergic neurons against it. Our results suggest that BH(4) functions as a complementary antioxidant to the glutathione/glutathione peroxidase system and that changes in BH(4) levels may contribute to the pathogenesis of PD. PMID- 10820191 TI - Chick pineal melatonin synthesis: light and cyclic AMP control abundance of serotonin N-acetyltransferase protein. AB - Melatonin production in the pineal gland is high at night and low during the day. This rhythm reflects circadian changes in the activity of serotonin N acetyltransferase [arylalkylamine N-acetyltransferase (AA-NAT); EC 2.3.1.87], the penultimate enzyme in melatonin synthesis. The rhythm is generated by an endogenous circadian clock. In the chick, a clock is located in the pinealocyte, which also contains two phototransduction systems. One controls melatonin production by adjusting the clock and the other acts distal to the clock, via cyclic AMP mechanisms, to switch melatonin synthesis on and off. Unlike the clock in these cells, cyclic AMP does not appear to regulate activity by altering AA NAT mRNA levels. The major changes in AA-NAT mRNA levels induced by the clock seemed likely (but not certain) to generate comparable changes in AA-NAT protein levels and AA-NAT activity. Cyclic AMP might also regulate AA-NAT activity via changes in protein levels, or it might act via other mechanisms, including posttranslational changes affecting activity. We measured AA-NAT protein levels and enzyme activity in cultured chick pineal cells and found that they correlated well under all conditions. They rose and fell spontaneously with a circadian rhythm. They also rose in response to agents that increase cyclic AMP. They were raised by agents that increase cyclic AMP, such as forskolin, and lowered by agents that decrease cyclic AMP, such as light and norepinephrine. Thus, both the clock and cyclic AMP can control AA-NAT activity by altering the total amount of AA-NAT protein. Effects of proteosomal proteolysis inhibitors suggest that changes in AA-NAT protein levels, in turn, reflect changes in the rate at which the protein is destroyed by proteosomal proteolysis. It is likely that cyclic AMP induced changes in AA-NAT protein levels mediate rapid changes in chick pineal AA NAT activity. Our results indicate that light can rapidly regulate the abundance of a specific protein (AA-NAT) within a photoreceptive cell. PMID- 10820192 TI - Increased expression of Galpha(q/11) and of phospholipase-Cbeta1/4 in differentiated human NT2-N neurons: enhancement of phosphoinositide hydrolysis. AB - The CNS is enriched in phosphoinositide-specific phospholipase C (PLC) and in the G proteins linked to its activation. Although the regional distributions of these signaling components within the brain have been determined, neither their cell type-specific localizations (i.e., neuronal versus glial) nor the functional significance of their high expression has been definitively established. In this study, we have examined the expression of phosphoinositide signaling proteins in human NT2-N cells, a well characterized model system for CNS neurons. Retinoic acid-mediated differentiation of NT2 precursor cells to the neuronal phenotype resulted in five- to 15-fold increases in the expression of PLC-beta1, PLC-beta4, and Galpha(q/11) (the prime G protein activator of these isozymes). In contrast, the expression of PLC-beta3 and PLC-gamma1 was markedly reduced following neuronal differentiation. Similar alterations in cell morphology and in the expression of PLC-beta1, PLC-beta3, and Galpha(q/11) expression were observed when NT2 cells were differentiated with berberine, a compound structurally unrelated to retinoic acid. NT2-N neurons exhibited a significantly higher rate of phosphoinositide hydrolysis than NT2 precursor cells in response to direct activation of either G proteins or PLC. These results indicate that neuronal differentiation of NT2 cells is associated with dramatic changes in the expression of proteins of the phosphoinositide signaling system and that, accordingly, differentiated NT2-N neurons possess an increased ability to hydrolyze inositol lipids. PMID- 10820193 TI - A monoclonal antibody to amyloid precursor protein induces neuronal apoptosis. AB - Although there is considerable evidence suggesting that altered metabolism of beta-amyloid precursor protein (APP) and accumulation of its beta-amyloid fragment are key features of Alzheimer's disease (AD), the normal physiological function of APP remains elusive. We investigated the potential role of APP in neurons using the monoclonal antibody 22C11, which binds to the extracellular domain of the human, rat, or mouse APP. Exposure of cortical neurons to 22C11 induced morphological changes including neurite degeneration, nuclear condensation, and internucleosomal DNA cleavage that were consistent with neurons dying by apoptosis. Supporting a role for 22C11-mediated apoptosis occurring by binding to APP were data demonstrating that preincubation of 22C11 with either purified APP or a synthetic peptide (APP(66-81)) that contains the epitope for 22C11 significantly attenuated neuronal damage induced by 22C11. The specificity of 22C11 was further supported by data showing no apparent effects of either mouse IgG or the monoclonal antibody P2-1, which is specific for the aminoterminal end of human but not rat APP. In addition, biochemical features indicative of apoptosis were the formation of 120- and 150-kDa breakdown products of fodrin following treatment of cortical neurons with 22C11. Both the morphological and the biochemical changes induced by 22C11 were prevented following pretreatment of neurons with the general caspase inhibitor N benzyloxycarbonyl-Val-Ala-Asp(O-methyl)-fluoromethyl ketone. Prior incubation of cortical neurons with GSH ethyl ester (GEE), a cell-permeable form of GSH, resulted in complete protection from the 22C11 insult, thus implicating an oxidative pathway in 22C11-mediated neuronal degeneration. This was further supported by the observation that prior treatment of neurons with buthionine sulfoximine, an inhibitor of gamma-glutamylcysteinyl synthetase, potentiated the toxic effects of 22C11. Finally, with use of compartmented cultures of hippocampal neurons, it was also demonstrated that selective application of 22C11 caused local neuritic degeneration that was prevented by the addition of GEE to the neuritic compartment. Thus, the binding of a monoclonal antibody to APP initially triggers neurite degeneration that is followed by caspase-dependent apoptosis in neuronal cultures and illustrates a novel property of this protein in neurons that may contribute to the profound neuronal cell death associated with AD. PMID- 10820194 TI - Unusual regulation of cyclin D1 and cyclin-dependent kinases cdk2 and cdk4 during in vivo mitotic stimulation of olfactory neuron progenitors in adult mouse. AB - The molecular mechanisms underlying cell cycle control in neuronal progenitors have been investigated with adult mouse olfactory epithelium as a model system. Odor receptive neurons of mammalian olfactory epithelium are short-lived and renewed in the adult by mitotic division of intrinsic neuronal progenitors. Ablation of the synaptic target, olfactory bulb, induces sequentially extensive apoptosis of sensory neurons and then stimulation of progenitor proliferation, peaking at 36 h and 4 days, respectively, postlesion. Known molecular effectors of G1 phase entry have been assessed on protein extracts of olfactory organs sampled at various postbulbectomy times in adult mice. The decay of betaIII tubulin and olfactory marker protein levels and the rise of proliferating cell nuclear antigen (PCNA) levels, starting 1 and 3 days, respectively, postlesion, provided the kinetic frame of neuronal dynamics. Cyclin D1, cyclin E, and cyclin dependent kinase cdk2 levels, low in olfactory organ of intact mice, increased 3 days after bulbectomy in parallel with PCNA levels; cdk4 content was initially high and unaffected by lesioning. Western blots of the known cdk inhibitors revealed proliferation-related decreases of p18, p21, and p27 from high expression in intact organs. Immunoprecipitation of cdk2 and cdk4 fractions of protein extracts at 4 days postlesion (mitotic reaction peak) versus control, followed by cyclin D1 immunoblotting, and vice versa, revealed that levels of both cyclin D1/cdk2 and cyclin D1/cdk4 complexes, as well as their kinase activities, were dramatically increased after lesion. In vivo proliferation of olfactory neuronal lineage cells thus involves functional binding of cyclin D1 with cdk2 and cdk4, with differential activation mechanisms for cdk2 and cdk4. In addition, the RT-PCR-detected cyclin D1 mRNA level remained unaffected after bulbectomy, which indicated that the cyclin D1 rise should involve posttranscriptional mechanisms in this in vivo neuronal system. These observations are discussed, along with their relevance to cell cycle control and to olfactory neuron dynamics. PMID- 10820195 TI - Spatiotemporal interaction of alpha(2) autoreceptors and noradrenaline transporters in the rat locus coeruleus: implications for volume transmission. AB - We investigated the roles of alpha(2) autoreceptors and noradrenaline (NA) transporters on NA efflux and uptake in the rat locus coeruleus after electrical stimulation. NA efflux was evoked by various trains (50 pulses, 10-500 Hz) and measured by fast cyclic voltammetry. NA efflux and uptake half-time (t(1/2)) were stimulus-dependent, ranging from 43 +/- 3 nM and 2.45 +/- 0.21 s, respectively, with 500-Hz stimuli to 127 +/- 11 nM and 4.41 +/- 0.34 s, respectively, with 100 Hz trains. Based on these data, we calculate that each transporter removes 0.19 NA molecules from the extracellular space every second, a velocity compatible more with transporter-than channel-mode conduction. Dexmedetomidine (10 nM) decreased NA efflux by approximately 30% on stimulations of < or =1 s in duration. BRL 44408 (1 microM) increased NA efflux on stimuli of > or =2 s (by up to 92 +/- 16%). Desipramine (50 nM) increased NA efflux on stimuli of > or =1 s (by 113 +/- 24%) but slowed NA uptake on all stimuli. When given together, the effects of desipramine and BRL 44408 were additive at stimuli of >or =1 s but showed potentiation on shorter trains. There was a significant time delay for the elevation of NA efflux by blockade of uptake (0.79 s) or autoreceptors (1.14 s), suggesting that both are located extrasynaptically and that NA must diffuse through the extracellular space to these structures. We suggest that released NA may interact with alpha(2) autoreceptors and NA transporters as far as 10 microm from the release sites, an action compatible with a volume transmission role of NA in the locus coeruleus. PMID- 10820196 TI - Effect of N-glycosylation on turnover and subcellular distribution of N acetylgalactosaminyltransferase I and sialyltransferase II in neuroblastoma cells. AB - Gangliosides are sialylated glycosphingolipids whose biosynthesis is catalyzed by a series of endoplasmic reticulum (ER)- and Golgi-resident glycosyltransferases. Protein expression, processing, and subcellular localization of the key regulatory enzymes for ganglioside biosynthesis, sialyltransferase II (ST-II) and N-acetylgalactosaminyltransferase I (GalNAcT), were analyzed upon transient expression of the two enzymes in the neuroblastoma cell lines NG108-15 and F-11. The enzymes were endowed with a C-terminal epitope tag peptide (FLAG) for immunostaining and immunoaffinity purification using a FLAG-specific antibody. Mature ST-II-FLAG and GalNAcT-FLAG were expressed as N-glycoproteins with noncomplex oligosaccharides. ST-II-FLAG was distributed to the Golgi apparatus, whereas GalNAcT-FLAG was found in the ER and Golgi. Inhibition of early N glycoprotein processing with castanospermine resulted in a distribution of ST-II FLAG to the ER, whereas that of GalNAcT-FLAG remained unaltered. In contrast to GalNAcT, the activity of ST-II and the amount of immunostained enzyme were reduced concomitantly by 75% upon incubation with castanospermine. This was due to a fourfold increased turnover of ST-II-FLAG, which was not found with GalNAcT FLAG. The ER retention and increased turnover of ST-II-FLAG were most likely due to its inability to bind to calnexin upon inhibition of early N-glycoprotein processing. Calnexin binding was not observed for GalNAcT-FLAG, indicating a differential effect of N-glycosylation on the turnover and subcellular localization of the two glycosyltransferases. PMID- 10820197 TI - Lithium ions enhance cysteine string protein gene expression in vivo and in vitro. AB - Lithium is a well established pharmacotherapy for the treatment of recurrent manic-depressive illness. However, the mechanism by which lithium exerts its therapeutic action remains elusive. Here we report that lithium at 1 mM significantly increased the expression of cysteine string proteins (CSPs) in a pheochromocytoma cell line (PC12 cells) differentiated by nerve growth factor. These cells concomitantly exhibited increased expression of CSPs in their cell bodies and boutons. Enhanced CSP expression was also observed in the brain of rats fed a lithium-containing diet, which elevated serum lithium to a therapeutically relevant concentration of approximately 1.0 mM. However, both in vitro and in vivo, the expression of another synaptic vesicle protein, synaptophysin, and the t-SNARE, synaptosomal-associated protein of 25 kDa (SNAP 25), was not significantly altered by lithium. These observations indicate that lithium-induced changes of CSP gene expression may contribute to the therapeutic efficacy of this monovalent cation. PMID- 10820198 TI - gp120 induces cell death in human neuroblastoma cells through the CXCR4 and CCR5 chemokine receptors. AB - To infect target cells, the human immunodeficiency virus (HIV) type I (HIV-1) must engage not only the well-known CD4 molecule, but it also requires one of several recently described coreceptors. In particular, the CXCR4 (LESTR/fusin) receptor allows fusion and entry of T-tropic strains of HIV, whereas CCR5 is the major coreceptor used by primary HIV-1 strains that infect macrophages and CD4(+) T-helper cells (M-tropic viruses). In addition, the alpha chemokine SDF1alpha and the beta chemokines MIP1alpha, MIP1beta, and RANTES, natural ligands of CXCR4 and CCR5, respectively, are potent soluble inhibitors of HIV infection by blocking the binding between the viral envelope glycoprotein gp120 and the coreceptors. Approximately two-thirds of individuals with acquired immunodeficiency syndrome (AIDS) show neurologic complications, which are referred to a syndrome called AIDS dementia complex or HIV-1-associated cognitive/motor complex. The HIV-1 coat glycoprotein gp120 has been proposed as the major etiologic agent for neuronal damage, mediating both direct and indirect effects on the CNS. Furthermore, recent findings showing the presence of chemokine receptors on the surface of different cell types resident in the CNS raise the possibility that the association of gp120 with these receptors may contribute to the pathogenesis of neurological dysfunction. Here, we address the possible role of alpha and beta chemokines in inhibiting gp120-mediated neurotoxicity using the human neuroblastoma CHP100 cell line as an experimental model. We have previously shown that, in CHP100 cells, picomolar concentrations of gp120 produce a significant increase in cell death, which seems to proceed through a Ca(2+) - and NMDA receptor-dependent cascade. In this study, we gained insight into the mechanism(s) of neurotoxicity elicited by the viral glycoprotein. We found that CHP100 cells constitutively express both CXCR4 and CCR5 receptors and that stimulation with phorbol 12-myristate 13-acetate down-regulates their expression, thus preventing gp120-induced cell death. Furthermore, all the natural ligands of these receptors exerted protective effects against gp120-mediated neuronal damage, although with different efficiencies. These findings, together with our previous reports, suggest that the neuronal injury observed in HIV-1 infection could be due to direct (or indirect) interactions between the viral protein gp120 and chemokine and/or NMDA receptors. PMID- 10820199 TI - Pregnenolone sulfate, a naturally occurring excitotoxin involved in delayed retinal cell death. AB - The present study was designed to investigate the neurosteroid pregnenolone sulfate (PS), known for its ability to modulate NMDA receptors and interfere with acute excitotoxicity, in delayed retinal cell death. Three hours after exposure of the isolated and intact retina to a 30-min PS pulse, DNA fragmentation as assessed by genomic DNA gel electrophoresis and a modified in situ terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling (TUNEL) method appeared concurrently with an increase in superoxide dismutase (SOD) activity and thiobarbituric acid-reactive substances (TBARS) levels. At 7 h, the increased amount of DNA laddering was accompanied by a higher number of TUNEL positive cells in the inner nuclear and ganglion cell layers. Necrotic signs were characterized by DNA smear migration, lactate dehydrogenase (LDH) release, and damage mainly in the inner nuclear layer. PS-induced delayed cell death was markedly reduced by the NMDA receptor antagonists 4-(3-phosphonopropyl)-2 piperazinecarboxylic acid and 3alpha-hydroxy-5beta-pregnan-20-one sulfate but completely blocked after concomitant addition of the non-NMDA receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione. Steroids with antioxidant properties (progesterone, dehydroepiandrosterone and its sulfate ester, and 17beta estradiol) differently prevented PS-induced delayed cell death. Cycloheximide treatment protected against DNA fragmentation and LDH release but failed to prevent the rise in SOD activity and TBARS level. We conclude that a brief PS pulse causes delayed cell death in a slowly evolving apoptotic fashion characterized by a cycloheximide-sensitive death program downstream of reactive oxygen species generation and lipid peroxidation, turning into secondary necrosis in a retinal cell subset. PMID- 10820200 TI - Transcriptional responses to growth factor and G protein-coupled receptors in PC12 cells: comparison of alpha(1)-adrenergic receptor subtypes. AB - Transcriptional responses to growth factor and G protein-coupled receptors were compared in PC12 cells using retroviral luciferase reporters. In cells stably expressing alpha(1A)-adrenergic receptors, norepinephrine activated all five reporters [AP1 (activator protein-1), SRE (serum response element), CRE (cyclic AMP response element), NFkappaB) (nuclear factor-kappaB), and NFAT (nuclear factor of activated T cells)], whereas nerve growth factor (NGF) and epidermal growth factor activated only AP1 and SRE. Activation of P2Y2 receptors by UTP did not activate any reporters. Protein kinase C inhibition blocked NFkappaB activation by norepinephrine, but potentiated CRE. Mitogen-activated protein kinase kinase inhibition blocked AP1 activation by norepinephrine, but also potentiated CRE. p38 mitogen-activated protein kinase inhibition reduced most norepinephrine responses, but not NGF responses. inhibition of Src eliminated SRE responses to norepinephrine and NGF, and reduced all responses except CRE. Phosphatidylinositol 3-kinase inhibitors markedly potentiated CRE activation by norepinephrine, with only small effects on the other responses. Comparison of the three human subtypes showed that the alpha(1A) activated all five reporters, the alpha(1B) showed smaller effects, and the alpha(1D) was ineffective. Cell differentiation caused by norepinephrine, but not NGF, was reduced by all inhibitors studied. These experiments suggest that alpha(1A)-adrenergic receptors activate a wider array of transcriptional responses than do growth factors in PC12 cells. These responses are not linearly related to second messenger production, and different subtypes show different patterns of activation. PMID- 10820201 TI - Kainic acid-induced neurotrophic activities in developing cortical neurons. AB - Using primary cultured cortical neurons from embryonic rat brains, we elucidated an alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA)/kainic acid (KA) receptor-mediated neuroprotective mechanism through actions of nerve growth factor (NGF) in developing neurons. Neurotoxicity of KA in early days in vitro neurons was quite low compared with the mature neurons. However, pretreatment with anti-NGF antibody or TrkA inhibitor AG-879 profoundly raised KA toxicity. Furthermore, KA stimulation resulted in an increase of TrkA expression and phosphorylation, which was blocked not only by the AMPA/KA receptor antagonist 6 cyano-7-nitroquinoxaline-2,3-dione and AG-879, but also by the phospholipase C inhibitor U73122 and the intracellular calcium chelator BAPTA. A study of polyphosphoinositide turnover showed that KA-stimulated phospholipase C (PLC) activity was directly triggered by the AMPA/KA receptor activity, but not by the activity of TrkA or other excitatory amino acid receptor subtypes. Sources of KA increased intracellular calcium levels were contributed by both extracellular calcium influx and intracellular calcium release and were partially sensitive to guanosine 5'-O-(2-thiodiphosphate). These results indicate that in developing cortical neurons, activation of AMPA/KA receptors by KA may induce expression, followed by activation of TrkA via PLC signaling and intracellular calcium elevation and hence increase reception of NGF on KA-challenged neurons. A G protein-coupled AMPA/KA receptor may be involved in these metabotropic events for neuronal protection. PMID- 10820202 TI - Retinoic acid-mediated enhancement of the cholinergic/neuronal nitric oxide synthase phenotype of the medial septal SN56 clone: establishment of a nitric oxide-sensitive proapoptotic state. AB - It is unclear what mechanisms lead to the degeneration of basal forebrain cholinergic neurons in Alzheimer's or other human brain diseases. Some brain cholinergic neurons express neuronal nitric oxide (NO) synthase (nNOS), which produces a free radical that has been implicated in some forms of neurodegeneration. We investigated nNOS expression and NO toxicity in SN56 cells, a clonal cholinergic model derived from the medial septum of the mouse basal forebrain. We show here that, in addition to expressing choline acetyltransferase (ChAT), SN56 cells express nNOS. Treatment of SN56 cells with retinoic acid (RA; 1 microM) for 48 h increased ChAT mRNA (+126%), protein (+88%), and activity (+215%) and increased nNOS mRNA (+98%), protein (+400%), and activity (+15%). After RA treatment, SN56 cells became vulnerable to NO excess generated with S nitro-N-acetyl-DL-penicillamine (SNAP) and exhibited increased nuclear DNA fragmentation that was blocked with a caspase-3 inhibitor. Treatment with dexamethasone, which largely blocked the RA-mediated increase in nNOS expression, or inhibition of nNOS activity with methylthiocitrulline strongly potentiated the apoptotic response to SNAP in RA-treated SN56 cells. Caspase-3 activity was reduced when SNAP was incubated with cells or cell lysates, suggesting that NO can directly inhibit the protease. Thus, whereas RA treatment converts SN56 cells to a proapoptotic state sensitive to NO excess, endogenously produced NO appears to be anti-apoptotic, possibly by tonically inhibiting caspase-3. PMID- 10820203 TI - Visualization and trafficking of the vesicular acetylcholine transporter in living cholinergic cells. AB - The present experiments investigated the trafficking of the vesicular acetylcholine transporter (VAChT) tagged with the enhanced green fluorescent protein (EGFP) in living cholinergic cells (SN56). The EGFP-VAChT chimera was located in endosomal-like compartments in the soma of SN56 cells, and it was also targeted to varicosities of neurites. In contrast, EGFP alone in cells was soluble in the cytoplasm. The C-terminal cytoplasmic tail of VAChT has been implicated in targeting of VAChT to synaptic vesicles; thus, we have examined the role of the C-terminal region in the trafficking to varicosities. A C-terminal fragment tagged with EGFP appeared to be selectively accumulated in varicosities when expressed in SN56 cells. Interestingly, the protein was not freely soluble in the cytosol, and it presented a punctate pattern of expression. However, EGFP C terminus did not present this peculiar pattern of expression in a nonneuronal cell line (HEK 293). Moreover, the C-terminal region of VAChT did not seem to be essential for VAChT trafficking, as a construct that lacks the C-terminal tail was, similar to EGFP-VAChT, partially targeted to endocytic organelles in the soma and sorted to varicosities. These experiments visualize VAChT for the first time in living cells and suggest that there might be multiple signals that participate in trafficking of VAChT to sites of synaptic vesicle accumulation. PMID- 10820204 TI - Light-induced tyrosine phosphorylation of BIT in the rat suprachiasmatic nucleus. AB - Circadian changes of protein tyrosine phosphorylation in the hypothalamic suprachiasmatic nucleus have been studied using rats maintained under 12-h light/ 12-h dark cycles as well as constant dark conditions. We found that tyrosine phosphorylation of BIT (brain immunoglobulin-like molecule with tyrosine-based activation motifs), a transmembrane glycoprotein of 90-95 kDa, was higher in the light period than in the dark period and was increased after light exposure in the dark period. Similar changes in tyrosine phosphorylation were observed under constant dark conditions, but its amplitude was weaker than that in 12-h light/12 h dark cycles. As the tyrosine-phosphorylated form of BIT is able to bind to the Src homology 2 domain of a protein tyrosine phosphatase, SHP-2, we examined association of these proteins in suprachiasmatic nucleus extracts and found that SHP-2 was coprecipitated with BIT in parallel with its tyrosine phosphorylation. These results suggest that tyrosine phosphorylation of BIT might be involved in light-induced entrainment of the circadian clock. PMID- 10820205 TI - GABA(A)-mediated toxicity of hippocampal neurons in vitro. AB - In the present study, we examined whether the elevation of GABA by gamma-vinyl GABA protects cultured rat fetal hippocampal neurons against toxicity induced by a 20-min incubation with 100 microM L-glutamate. Neither a 24-h pretreatment nor posttreatment with gamma-vinyl-GABA (100 microM) had any neuroprotective effects, as determined by counting microtubule-associated protein-2 positive cells and lactate dehydrogenase assay 24 h after the glutamate treatment. Unexpectedly, gamma-vinyl-GABA alone induced a 20% loss of microtubule-associated protein-2 positive cells in a culture that was grown in medium containing 25 mM KCl. The toxic effect of gamma-vinyl-GABA was mimicked by a 24-h treatment with GABA (100 microM) and the GABA(A) receptor agonist, muscimol (10 microM), but not the GABA(B) receptor agonist, baclofen (10 microM). The GABA(A) receptor antagonist, bicuculline (10 microM), protected against gamma-vinyl-GABA and GABA-evoked toxicity. Neither gamma-vinyl-GABA nor GABA was toxic in culture medium containing 15 mM KCl. These data indicate that, under depolarizing conditions, an increased GABA level is toxic for a subpopulation of developing hippocampal neurons in vitro. The effect is GABA(A) receptor-mediated. These data provide a new view for understanding neurodegenerative processes, and raise a question of the safety of therapies aimed at increasing GABA concentration following brain insults, especially in immature brains. PMID- 10820206 TI - Synergetic activation of p38 mitogen-activated protein kinase and caspase-3-like proteases for execution of calyculin A-induced apoptosis but not N-methyl-d aspartate-induced necrosis in mouse cortical neurons. AB - We examined the possibility that p38 mitogen-activated protein kinase and caspase 3 would be activated for execution of apoptosis and excitotoxicity, the two major types of neuronal death underlying hypoxicischemic and neurodegenerative diseases. Mouse cortical cell cultures underwent widespread neuronal apoptosis 24 h following exposure to 10-30 nM calyculin A, a selective inhibitor of Ser/Thr phosphatase I and IIA. Activity of p38 was increased 2-4 h following exposure to 30 nM calyculin A. Addition of 3-10 microM PD169316, a selective p38 inhibitor, partially attenuated calyculin A neurotoxicity. Activity of caspase-3-like proteases was increased in cortical cell cultures exposed to 30 nM calyculin A for 8-16 h as shown by cleavage of DEVD-p-nitroanilide and phosphorylated tau. Proteolysis of tau was completely blocked by addition of 100 microM N benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone (z-VAD-fmk), a broad-spectrum inhibitor of caspases, but incompletely by 10 microM PD169316. Calyculin A neurotoxicity was partially sensitive to 100 microM z-VAD-fmk. Cotreatment with 10 microM PD169316 and 100 microM z-VAD-fmk showed additive neuroprotection against calyculin A. Neither PD169316 nor z-VAD-fmk showed a beneficial effect against excitotoxic neuronal necrosis induced by exposure to 20 microM NMDA. Thus, caspase-3-like proteases and p38 likely contribute to calyculin A-induced neuronal apoptosis but not NMDA-induced neuronal necrosis. PMID- 10820207 TI - A(2A) adenosine receptor facilitation of neuromuscular transmission: influence of stimulus paradigm on calcium mobilization. AB - The influence of stimulus pulse duration on calcium mobilization triggering facilitation of evoked [(3)H]acetylcholine ([(3)H]ACh) release by the A(2A) adenosine receptor agonist CGS 21680C was studied in the rat phrenic nerve hemidiaphragm. The P-type calcium channel blocker omega-agatoxin IVA (100 nM) decreased [(3)H]ACh release evoked with pulses of 0.04-ms duration, whereas nifedipine (1 microM) inhibited transmitter release with pulses of 1-ms duration. Depletion of intracellular calcium stores by thapsigargin (2 microM) decreased [(3)H]ACh release evoked by pulses of 1 ms, an effect observed even in the absence of extracellular calcium. With short (0.04-ms) stimulation pulses, when P type calcium influx triggered transmitter release, facilitation of [(3)H]ACh release by CGS 21680C (3 nM) was attenuated by both thapsigargin (2 microM) and nifedipine (1 microM). With longer stimuli (1 ms), a situation in which both thapsigargin-sensitive internal stores and L-type channels are involved in ACh release, pretreatment with either omega-agatoxin IVA (100 nM) or nifedipine (1 microM) reduced the facilitatory effect of CGS 21680C (3 nM). The results suggest that A(2A) receptor activation facilitates ACh release from motor nerve endings through alternatively mobilizing the available calcium pools (thapsigargin sensitive internal stores and/or P- or L-type channels) that are not committed to the release process in each stimulation condition. PMID- 10820208 TI - Regulation of tyrosine hydroxylase activity and phosphorylation at Ser(19) and Ser(40) via activation of glutamate NMDA receptors in rat striatum. AB - The activity of tyrosine hydroxylase, the rate-limiting enzyme in the biosynthesis of dopamine, is stimulated by phosphorylation. In this study, we examined the effects of activation of NMDA receptors on the state of phosphorylation and activity of tyrosine hydroxylase in rat striatal slices. NMDA produced a time-and concentration-dependent increase in the levels of phospho Ser(19)-tyrosine hydroxylase in nigrostriatal nerve terminals. This increase was not associated with any changes in the basal activity of tyrosine hydroxylase, measured as DOPA accumulation. Forskolin, an activator of adenylyl cyclase, stimulated tyrosine hydroxylase phosphorylation at Ser(40) and caused a significant increase in DOPA accumulation. NMDA reduced forskolin-mediated increases in both Ser(40) phosphorylation and DOPA accumulation. In addition, NMDA reduced the increase in phospho-Ser(40)-tyrosine hydroxylase produced by okadaic acid, an inhibitor of protein phosphatase 1 and 2A, but not by a cyclic AMP analogue, 8-bromo-cyclic AMP. These results indicate that, in the striatum, glutamate decreases tyrosine hydroxylase phosphorylation at Ser(40) via activation of NMDA receptors by reducing cyclic AMP production. They also provide a mechanism for the demonstrated ability of NMDA to decrease tyrosine hydroxylase activity and dopamine synthesis. PMID- 10820209 TI - Analyses of signal transduction cascades reveal an essential role of calcium ions for regulation of melatonin biosynthesis in the light-sensitive pineal organ of the rainbow trout (Oncorhynchus mykiss). AB - Signal transduction processes regulating melatonin production in the light sensitive trout pineal organ were investigated by immunocytochemical and immunochemical demonstration of phosphorylated cyclic AMP-responsive element binding protein (pCREB) and measurements of cyclic AMP, melatonin, and calcium levels. Melatonin levels were tightly controlled by light and darkness. Elevation of cyclic AMP levels by 8-bromo-cyclic AMP, forskolin, and 3-isobutyl-1 methylxanthine increased the levels of pCREB and melatonin in light- or dark adapted pineal organs in vitro. Without pharmacological treatment, the levels of pCREB and cyclic AMP remained constant for several hours before and after light onset. Inhibition of cyclic AMP-dependent proteasomal proteolysis by lactacystin, MG 132, and calpain inhibitor I did not prevent the rapid, light-induced suppression of melatonin biosynthesis. However, changes in the intracellular calcium concentration by drugs affecting voltage-gated calcium channels of the L type and intracellular calcium oscillations (cobalt chloride, nifedipine, Bay K 8644) had dramatic effects on the rapid, light-dependent changes in melatonin levels. These effects were not accompanied by changes in cyclic AMP levels. Thus, the rapid, light-dependent changes in melatonin levels in the trout pineal organ are regulated apparently by a novel calcium signaling pathway and do not involve changes in cyclic AMP levels, cyclic AMP-dependent proteasomal proteolysis, or phosphorylation of cyclic AMP-responsive element-binding protein. PMID- 10820210 TI - Nongenomic inhibition of catecholamine secretion by 17beta-estradiol in PC12 cells. AB - We investigated the effects of 17beta-estradiol, an estrogen, on [(3)H]norepinephrine ([(3)H]NE) secretion in PC12 cells. Pretreatment with 17beta estradiol reduced 70 mM K(+)-induced [(3)H]NE secretion in a concentration dependent manner with a half-maximal inhibitory concentration (IC(50)) of 2 +/- 1 microM. The 70 mM K(+)-induced cytosolic free Ca(2+) concentration ([Ca(2+)](i)) rise was also reduced when the cells were treated with 17beta-estradiol (IC(50) = 15 +/- 2 microM). Studies with voltage-sensitive calcium channel (VSCC) antagonists such as nifedipine and omega-conotoxin GVIA revealed that both L- and N-type VSCCs were affected by 17beta-estradiol treatment. The 17beta-estradiol effect was not changed by pretreatment of the cells with actinomycin D and cycloheximide for 5 h. In addition, treatment with pertussis or cholera toxin did not affect the inhibitory effect of 17beta-estradiol. 17beta-Estradiol also inhibited the ATP-induced [(3)H]NE secretion and [Ca(2+)](i) rise. In PC12 cells, the ATP-induced [Ca(2+)](i) rise is known to occur through P2X(2) receptors, the P2Y(2)-mediated phospholipase C (PLC) pathway, and VSCCs. 17beta-Estradiol pretreatment during complete inhibition of the PLC pathway and VSCCs inhibited the ATP-induced [Ca(2+)](i) rise. Our results suggest that 17beta-estradiol inhibits catecholamine secretion by inhibiting L- and N-type Ca(2+) channels and P2X(2) receptors in a nongenomic manner. PMID- 10820211 TI - Somatostatin release in the hippocampus in the kindling model of epilepsy: a microdialysis study. AB - Somatostatin biosynthesis in the hippocampus is activated during and following kindling epileptogenesis. The aim of this study was to investigate whether this phenomenon is associated with enhanced somatostatin release in vivo. Experiments have been run in awake, freely moving rats, implanted with a bipolar electrode in the right amygdala (for kindling stimulation), and with a recording electrode and a microdialysis probe in the left hippocampus. Basal somatostatin-like immunoreactivity (-LI) release was significantly greater in kindled than naive rats. In naive rats, a 2-min perfusion with 100 mM K(+) did not affect behavior and EEG recordings and nonsignificantly increased somatostatin-LI release; a 10 min K(+) perfusion evoked numerous wet dog shakes, electrical seizures (class 0; latency congruent with 8 min, duration congruent with 8 min), and somatostatin-LI release ( congruent with 350% of basal); and a single kindling after-discharge (4 +/- 3-s duration in the hippocampus) also evoked somatostatin-LI release ( congruent with 200% of basal). In kindled rats, a 2-min 100 mM K(+) perfusion evoked hippocampal discharges in three of seven animals (latency congruent with 2 min, mean duration congruent with 1.5 min) and increased somatostatin-LI release ( congruent with 250% of basal); a 10-min K(+) perfusion evoked behavioral seizures (class 1 to 5, latency congruent with 4 min, mean duration congruent with 12 min) with numerous wet dog shakes and robust somatostatin-LI release ( congruent with 350% of basal); and a kindling stimulation evoked generalized seizures (class 4 or 5, 77 +/- 15-s duration in the hippocampus) with remarkable somatostatin-LI release ( congruent with 300% of basal). These data demonstrate that hippocampal somatostatin release is increased in the kindling model in vivo. PMID- 10820212 TI - Poly(ADP-ribose) synthase inhibition reduces ischemic injury and inflammation in neonatal rat brain. AB - Poly(ADP-ribose) synthase (PARS), an abundant nuclear protein, has been described as an important candidate for mediation of neurotoxicity by nitric oxide. However, in cerebral ischemia, excessive PARS activation may lead to energy depletion and exacerbation of neuronal damage. We examined the effect of inhibiting PARS on the (a) degree of cerebral injury, (b) process of inflammatory responses, and (c) functional outcomes in a neonatal rat model of focal ischemia. We demonstrate that administration of 3-aminobenzamide, a PARS inhibitor, leads to a significant reduction of infarct volume: 63 +/- 2 (untreated) versus 28 +/- 4 mm(3) (treated). The neuroprotective effects currently observed 48 h postischemia hold up at 7 and 17 days of survival time and attenuate neurological dysfunction. Inhibition of PARS activity, demonstrated by a reduction in poly(ADP ribose) polymer formation, also reduces neutrophil recruitment and levels of nitrotyrosine, an indicator of peroxynitrite generation. Taken together, our results demonstrate that PARS inhibition reduces ischemic damage and local inflammation associated with reperfusion and may be of interest for the treatment of neonatal stroke. PMID- 10820213 TI - Accumulation of N-acetyl-L-aspartate in the brain of the tremor rat, a mutant exhibiting absence-like seizure and spongiform degeneration in the central nervous system. AB - The tremor rat is a mutant that exhibits absence-like seizure and spongiform degeneration in the CNS. By positional cloning, a genomic deletion was found within the critical region in which the aspartoacylase gene is located. Accordingly, no aspartoacylase expression was detected in any of the tissues examined, and abnormal accumulation of N-acetyl-L-aspartate (NAA) was shown in the mutant brain, in correlation with the severity of the vacuole formation. Therefore, the tremor rat may be regarded as a suitable animal model of human Canavan disease, characterized by spongy leukodystrophy that is caused by aspartoacylase deficiency. Interestingly, direct injection of NAA into normal rat cerebroventricle induced 4- to 10-Hz polyspikes or spikewave-like complexes in cortical and hippocampal EEG, concomitantly with behavior characterized by sudden immobility and staring. These results suggested that accumulated NAA in the CNS would induce neuroexcitation and neurodegeneration directly or indirectly. PMID- 10820214 TI - Oxidative modification of creatine kinase BB in Alzheimer's disease brain. AB - Creatine kinase (CK) BB, a member of the CK gene family, is a predominantly cytosolic CK isoform in the brain and plays a key role in regulation of the ATP level in neural cells. CK BB levels are reduced in brain regions affected by neurodegeneration in Alzheimer's disease (AD), Pick's disease, and Lewy body dementia, and this reduction is not a result of decreased mRNA levels. This study demonstrates that posttranslational modification of CK BB plays a role in the decrease of CK activity in AD brain. The specific CK BB activity and protein carbonyl content were determined in brain extracts of six AD and six age-matched control subjects. CK BB activity per microgram of immunoreactive CK BB protein was lower in AD than in control brain extracts, indicating the presence of inactive CK BB molecules. The analysis of specific protein carbonyl levels in CK BB, performed by two-dimensional fingerprinting of oxidatively modified proteins, identified CK BB as one of the targets of protein oxidation in the AD brain. The increase of protein carbonyl content in CK BB provides evidence that oxidative posttranslational modification of CK BB plays a role in the loss of CK BB activity in AD. PMID- 10820215 TI - Enhancement of diphtheria toxin potency by replacement of the receptor binding domain with tetanus toxin C-fragment: a potential vector for delivering heterologous proteins to neurons. AB - This study describes the expression, purification, and characterization of a recombinant fusion toxin, DAB(389)TTC, composed of the catalytic and membrane translocation domains of diphtheria toxin (DAB(389)) linked to the receptor binding fragment of tetanus toxin (C-fragment). As determined by its ability to inhibit cellular protein synthesis in primary neuron cultures, DAB(389)TTC was approximately 1,000-fold more cytotoxic than native diphtheria toxin or the previously described fusion toxin, DAB(389)MSH. The cytotoxic effect of DAB(389)TTC on cultured cells was specific toward neuronal-type cells and was blocked by coincubation of the chimeric toxin with tetanus antitoxin. The toxicity of DAB(389)TTC, like that of diphtheria toxin, was dependent on passage through an acidic compartment and ADP-ribosyltransferase activity of the DAB(389) catalytic fragment. These results suggest that a catalytically inactive form of DAB(389)TTC may be useful as a nonviral vehicle to deliver exogenous proteins to the cytosolic compartment of neurons. PMID- 10820216 TI - Neuronal nicotinic acetylcholine receptors from Drosophila: two different types of alpha subunits coassemble within the same receptor complex. AB - Although neuronal nicotinic acetylcholine receptors from insects have been reconstituted in vitro more than a decade ago, our knowledge about the subunit composition of native receptors as well as their functional properties still remains limited. Immunohistochemical evidence has suggested that two alpha subunits, alpha-like subunit (ALS) and Drosophila alpha2 subunit (Dalpha2), are colocalized in the synaptic neuropil of the Drosophila CNS and therefore may be subunits of the same receptor complex. To gain further understanding of the composition of these nicotinic receptors, we have examined the possibility that a receptor may imbed more than one alpha subunit using immunoprecipitations and electrophysiological investigations. Immunoprecipitation experiments of fly head extracts revealed that ALS-specific antibodies coprecipitate Dalpha2, and vice versa, and thereby suggest that these two alpha subunits must be contained within the same receptor complex, a result that is supported by investigations of reconstituted receptors in Xenopus oocytes. Discrimination between binary (ALS/beta2 or Dalpha2/beta2) and ternary (ALS/Dalpha2/beta2) receptor complexes was made on the basis of their dose-response curve to acetylcholine as well as their sensitivity to alpha-bungarotoxin or dihydro-beta-erythroidine. These data demonstrate that the presence of the two alpha subunits within a single receptor complex confers new receptor properties that cannot be predicted from knowledge of the binary receptor's properties. PMID- 10820217 TI - Purification and structure of neurostatin, an inhibitor of astrocyte division of mammalian brain. AB - Neurostatin was originally described as an inhibitor of astroblast and astrocytoma division present in rat brain extracts and immunologically related to the sugar moiety of epidermal growth factor receptor and to blood group antigens. It was purified recently from mammalian brain extracts and characterized as a glycosphingolipid, but its precise structure remained unknown. Neurostatin has now been purified to apparent homogeneity from ganglioside extracts of rat, bovine, and porcine brain. It is cytostatic for astroblasts, C6 glioma cells, and various human astrocytomas grades III and IV, with IC(50) values ranging from 250 to 450 nM, but does not affect the division of primary or transformed fibroblasts up to concentrations >4 microM. Matrix-assisted laser desorption ionization time of-flight mass spectrometry of purified pig neurostatin showed a molecular ion of 1, 905 Da and ions of 1,863 and 1,934 Da, compatible with a disialoganglioside. Mono- and bidimensional NMR spectra, together with biochemical studies, suggest that neurostatin may be the 9-O-monoacetyl ester of GD1b. PMID- 10820218 TI - Identification of the isoforms of Ca(2+)/Calmodulin-dependent protein kinase II in rat astrocytes and their subcellular localization. AB - Ca(2+)/calmodulin-dependent protein kinase II (CaM kinase II) occurs in astrocytes as well as in neurons in brain. We have reported that CaM kinase II is involved in the regulation of cytoskeletal proteins and gene expression in astrocytes. In this study, we identified all isoforms of CaM kinase II in astrocytes and examined their subcellular localization. When we amplified the isoforms of four subunits by RT-PCR followed by the "nested" PCR, totally 10 isoforms were obtained. Immunoblot analyses with five types of antibodies against CaM kinase II indicated that the most abundant isoform was delta2. Immunostaining suggested that the delta2 isoform was localized predominantly at the Golgi apparatus. The localization of the delta2 isoform at the Golgi apparatus was also observed in NG108-15 cells. We overexpressed all isoforms that contained the nuclear localization signal to examine their nuclear targeting in NG108-15 cells. In contrast to the alphaB and delta3 isoforms that entered the nucleus, as reported, the gammaA isoform was excluded from the nucleus in the transfected NG108-15 cells. These results suggest that the 15-amino acid insertion following the nuclear localization signal inhibits the nuclear targeting of the gammaA isoform. PMID- 10820219 TI - Effect of stress exposure on the activation pattern of enkephalin-containing perikarya in the rat ventral medulla. AB - We examined the effects of acute and chronic psychogenic stress on the activation pattern of enkephalin-containing perikarya in the rat ventrolateral medulla. Rats allocated to the chronic stress groups were subjected to 90 min of immobilization for 10 days. On the 11th day, the chronically stressed rats were exposed to homotypic (90-min immobilization) or to heterotypic but still psychogenic (90-min immobilization coupled to air jet stress) stress. The acute stress group was subjected once to an acute 90-min immobilization. For each group, the rats were anesthetized either before stress (time 0) or 90, 180, and 270 min after the onset of stress. Brain sections were then processed using immunocytochemistry (Fos protein) followed by radioactive in situ hybridization histochemistry (enkephalin mRNA). Following immobilization, the acute group displayed a marked increase in the number of activated enkephalin-containing perikarya within the paragigantocellularis and lateral reticular nuclei. This level of activation was sustained up to 180 min following the onset of the immobilization stress and had returned to baseline levels by 270 min from the initiation of the stress. However, this stress-induced activation of enkephalin-containing perikarya of the ventrolateral medulla was not seen following either homotypic or heterotypic stress in the chronically stressed group. These results provide evidence that enkephalin-containing perikarya of the ventrolateral medulla may constitute a potential circuit through which they regulate some aspect of the stress responses. Conversely, this enkephalinergic influence from the ventrolateral medulla was shown to be absent following chronic stress exposure. This would suggest a decrease in enkephalin inhibitory input originating from the ventrolateral medulla, thereby allowing a neuroendocrine and/or autonomic response to chronic stress. PMID- 10820220 TI - Regulatory effects of D2 receptors in the ventral tegmental area on the mesocorticolimbic dopaminergic pathway. AB - To investigate the regulatory effects of somatodendritic D2 receptors on the terminal's extracellular dopamine (DA) concentration, a D2 antagonist (eticlopride) was infused directly into the ventral tegmental area via a microdialysis probe in chloral hydrate-anesthetized rats. Extracellular DA changes in both the nucleus accumbens (N ACC) and the medial prefrontal cortex (mPFC) were monitored. Infusion of 10.0 fM eticlopride had no effect on DA in the mPFC (110.2 +/- 10.0% of baseline) but significantly increased DA in the N ACC (150.1 +/- 11.7%). Infusion of a higher dose of eticlopride (100.0 or 1,000.0 fM) significantly augmented the DA in the mPFC (121.1 +/- 7.6 and 180.7 +/- 25.8%, respectively) but surprisingly had no effect on DA in the N ACC (111.5 +/- 7.3 and 104.1 +/- 8.7%, respectively). To further investigate whether the bluntness of DA increase in the N ACC was due to DA receptor activation in the mPFC, eticlopride or SCH23390 was infused into the mPFC prior to and during intrategmental eticlopride infusion, and the change of DA in the N ACC was simultaneously monitored. During intra-mPFC 1.0 nM eticlopride infusion but not during 10.0 nM SCH23390 administration (95.5 +/- 6.1%), intrategmental 1,000.0 fM eticlopride infusion could further elevate DA in the N ACC (130.0 +/- 4.6%). Our results indicated that (1) the mesolimbic and the mesocortical pathways were under tonic inhibition by somatodendritic D2 receptors; (2) the DA concentration in the N ACC first increased and then returned to baseline while the intrategmental infusion dose of eticlopride increased; and (3) the bluntness of DA increase in the N ACC resulted from the D2 receptor activation in the mPFC. PMID- 10820221 TI - The FTDP-17-linked mutation R406W abolishes the interaction of phosphorylated tau with microtubules. AB - The recent finding that several point mutations in the gene encoding for the microtubule-binding protein tau correlate with neurological disorders has heightened interest in the mechanisms of destabilization of this protein. In this study the functional consequences of the tau mutation R406W on the interaction of the protein with microtubules have been analyzed. Mutated tau is less phosphorylated than its normal counterpart at serines 396 and 404. Furthermore, the phosphorylated mutant protein is unable to bind to microtubules, and, as a consequence, microtubules assembled after transient nocodazole treatment in the presence of this tau variant contain only unmodified tau and appear to form more and longer bundles than those assembled in the presence of wild-type tau. We propose that phosphorylated tau, unbound to microtubules, could accumulate in the cytoplasm. PMID- 10820222 TI - Electrophysiological evidence for the coexistence of alpha1 and alpha6 subunits in a single functional GABA(A) receptor. AB - The subunit combinations alpha1beta2gamma2, alpha6beta2gamma2, and alpha1alpha6beta2gamma2 of the GABA(A) receptor were functionally expressed in Xenopus oocytes. The properties of the resulting ion currents were characterized by using electrophysiological techniques. The concentration-response curve of the channel agonist GABA for alpha1alpha6beta2gamma2 showed a single apparent component characterized by an EC(50) of 107 +/- 26 microM (n = 4). It was different from the one for alpha1beta2gamma2, which had an EC(50) of 41 +/- 9 microM (n = 4), that for alpha6beta2gamma2, with an EC(50) of 6.7 +/- 1.9 microM (n = 5), and those for alpha1beta2 and alpha1alpha6beta2. There was no appreciable functional expression of alpha6beta2. Allosteric responses of alpha1alpha6beta2gamma2 to diazepam were intermediate to those of alpha1beta2gamma2 and alpha6beta2gamma2, and allosteric responses to flumazenil were comparable to the ones for alpha1beta2gamma2. The inhibition by furosemide of the currents elicited by GABA in alpha1alpha6beta2gamma2 [IC(50) = 298 +/- 116 microM (n = 7), assuming only one component] was not identical with inhibition of alpha6beta2gamma2 (IC(50) = 38 +/- 2 microM, n = 4), alpha1beta2gamma2 (IC(50) = 5,610 +/- 910 microM, n = 5), or a mixture of these components (assuming two components). These findings indicate unambiguously the formation of functional GABA(A) receptors containing two different alpha subunits, alpha1 and alpha6, with properties different from those of alpha1beta2gamma2 and alpha6beta2gamma2. Furthermore, we provide evidence for the facts that in the Xenopus oocyte (a) the formation of the different receptor types depends on the relative abundance of cRNAs coding for the different receptor subunits and (b) that functional dual subunit combinations alphabeta do not form in the presence of cRNA coding for the gamma subunit. PMID- 10820223 TI - Structure-activity relationships among N-arachidonylethanolamine (Anandamide) head group analogues for the anandamide transporter. AB - Two putative endocannabinoids, N-arachidonylethanolamine (AEA) and 2 arachidonylglycerol, are inactivated by removal from the extracellular environment by a process that has the features of protein-mediated facilitated diffusion. We have synthesized and studied 22 N-linked analogues of arachidonylamide for the purpose of increasing our understanding of the structural requirements for the binding of ligands to the AEA transporter. We have also determined the affinities of these analogues for both the CB(1) cannabinoid receptor and fatty acid amide hydrolase (FAAH). We have identified several structural features that enhance binding to the AEA transporter in cerebellar granule cells. We have confirmed the findings of others that replacing the ethanolamine head group with 4-hydroxybenzyl results in a high-affinity ligand for the transporter. However, we find that the same molecule is also a competitive inhibitor of FAAH. Similarly, replacement of the ethanolamine of AEA with 3-pyridinyl also results in a high-affinity inhibitor of both the transporter and FAAH. We conclude that the structural requirements for ligand binding to the CB(1) receptor and binding to the transporter are very different; however, the transporter and FAAH share most, but not all, structural requirements. PMID- 10820224 TI - A synthetic peptide ligand of neural cell adhesion molecule (NCAM) IgI domain prevents NCAM internalization and disrupts passive avoidance learning. AB - The neural cell adhesion molecule (NCAM) mediates cell adhesion and signal transduction through trans-homophilic- and/or cis-heterophilic-binding mechanisms. Intraventricular infusions of anti-NCAM have revealed a functional requirement of NCAM for the consolidation of memory in rats and chicks in a specific interval 6-8 h after training. We have now extended these studies to a synthetic peptide ligand of NCAM (C3) with an affinity for the IgI domain and the capability of inhibiting NCAM-mediated neurite outgrowth in vitro. Intraventricular administration of a single 5 microg bolus of C3 strongly inhibited recall of a passive avoidance response in adult rats, when given during training or in the 6-8-h posttraining period. The effect of C3 on memory consolidation was similar to that obtained with anti-NCAM as the amnesia was not observed until the 48-h recall time. The unique amnesic action of C3 during training could be related to disrupted NCAM internalization following training. In the 3-4-h posttraining period NCAM 180, the synapse-associated isoform, was down-regulated in the hippocampal dentate gyrus. This effect was mediated by ubiquitination and was prevented by C3 administration during training. These findings indicate NCAM to be involved in both the acquisition and consolidation of a passive avoidance response in the rat. Moreover, the study provides the first in vivo evidence for NCAM internalization in learning and identifies a synthetic NCAM ligand capable of modulating memory processes in vivo. PMID- 10820225 TI - Taurine blocks spontaneous cone contraction but not horizontal cell dark suppression in isolated goldfish retina. AB - The objective of this study was to investigate the effects of taurine on cone retinomotor movements and the responses of cone-driven horizontal cells in dark adapted teleost retina. In isolated goldfish retina preparations maintained in the dark, cones spontaneously contracted, and the responses of horizontal cells were suppressed. Addition of 5 mM taurine to the physiological solution blocked the spontaneous contraction of cones in the dark but did not block the dark suppression of horizontal cell responses. These results indicate that the mechanism that leads to horizontal cell dark suppression is not sensitive to taurine. Although both cone retinomotor position and horizontal cell responsiveness are known to be modulated by dopamine, the present results do not support the hypothesis that taurine inhibits dopamine release in the dark because only spontaneous cone contraction was affected by taurine. These results also indicate that spontaneous cone contraction in the dark is not the cause of horizontal cell dark suppression because, in the presence of taurine, cones were elongated yet horizontal cell responses were still suppressed. Consequently, these results make it clear that horizontal cell dark suppression is not an artifact produced by incubating isolated teleost retina preparations in taurine free physiological solution. PMID- 10820226 TI - Molecular cloning of a novel brain-type Na(+)-dependent inorganic phosphate cotransporter. AB - We have isolated a human cDNA encoding a protein, designated DNPI, that shows 82% amino acid identity and 92% similarity to the human brain-specific Na(+) dependent inorganic phosphate (Na(+)/P(i)) cotransporter (BNPI), which is localized exclusively to neuron-rich regions. Expression of DNPI mRNA in Xenopus oocytes resulted in a significant increase in Na(+)-dependent P(i) transport, indicating that DNPI is a novel Na(+)/P(i) cotransporter. Northern blot analysis shows that DNPI mRNA is expressed predominantly in brain, where the highest levels are observed in medulla, substantia nigra, subthalamic nucleus, and thalamus, all of which express BNPI mRNA at low levels. In contrast, DNPI mRNA is expressed at low levels in cerebellum and hippocampus, where BNPI mRNA is expressed at high levels. No hybridizing signal for DNPI mRNA is observed in the glia-rich region of corpus callosum. In other regions examined, both mRNAs are moderately or highly expressed. These results indicate that BNPI and DNPI, which coordinate Na(+)-dependent P(i) transport in the neuron-rich regions of the brain, may form a new class within the Na(+)/P(i) cotransporter family. PMID- 10820227 TI - Cutting edge: persistent fetal microchimerism in T lymphocytes is associated with HLA-DQA1*0501: implications in autoimmunity. AB - The host's MHC genotype plays a critical role in susceptibility to autoimmune diseases. We previously proposed that persistent fetal microchimerism from pregnancy contributes to the pathogenesis of autoimmune diseases such as scleroderma. In the current study, we investigated whether the specific host MHC genotype is associated with persistent microchimerism among T lymphocytes in women with scleroderma and in healthy women. Fetal microchimerism among T lymphocytes was strongly associated with HLA DQA1*0501 of the mother (odds ratio (OR) = 13.5, p = 0.007, p corrected (pc) = 0.06) and even more strongly with DQA1*0501 of the son (OR = infinity; p = 0. 00002, pc = 0.0002). This is the first description of an association between persistent fetal microchimerism in maternal T lymphocytes and specific HLA class II alleles. Although the association was observed in both healthy women and in women with scleroderma, the finding suggests an additional route by which HLA genes might contribute to susceptibility to autoimmune disease. PMID- 10820228 TI - Murine bone marrow-derived mast cells as potent producers of IL-9: costimulatory function of IL-10 and kit ligand in the presence of IL-1. AB - Recently, the Th2-type cytokine IL-9 was identified by genetic mapping analyses as a key mediator that determines the susceptibility to asthma. This has been further supported by data from IL-9-transgenic mice in which the overexpression of IL-9 in the lung causes airway inflammation, mast cell hyperplasia, and bronchial hyperresponsiveness. In an accompanying paper, we demonstrate that murine bone marrow-derived mast cells (BMMC) after stimulation with either ionomycin, a combination of ionomycin and IL-1, or via IgE-Ag complexes and IL-1 are very potent producers of IL-9. Herein we show that a dramatic increase of IL 9 production is observed when BMMC activated with ionomycin/IL-1 or with IgE-Ag complexes/IL-1 are treated with either additional kit ligand (KL) or IL-10. Both KL and IL-10 considerably enhance the production of IL-9 mRNA and protein. We were also able to demonstrate that the production of endogenous IL-10 by activated mast cells acts on the production of IL-9. Half-life measurements of IL 9 mRNA revealed no significant effect by KL, but a 2-fold increase of mRNA stability under the influence of IL-10. Reporter gene assays of transfected BMMC showed an enhanced transcriptional activity of the IL-9 promoter in the presence of either IL-10 or KL compared with cells stimulated only with a combination of IL-1 and ionomycin. The influence of KL and IL-10 might be of physiological importance, because it is known that both cytokines are produced by bronchial epithelial cells. PMID- 10820229 TI - In activated mast cells, IL-1 up-regulates the production of several Th2-related cytokines including IL-9. AB - Mast cells can play detrimental roles in the pathophysiology and mortality observed in anaphylaxis and other Th2-dominated allergic diseases. In contrast, these cells contribute to protective host defense mechanisms against parasitic worm infections. After IgE/Ag activation, mast cells can produce multiple cytokines that may enhance allergic inflammations, while a similar panel of Th2 related cytokines may support immunological strategies against parasites. Here we report that in primary mouse bone marrow-derived mast cells activated by ionomycin or IgE/Ag, the proinflammatory mediator IL-1 (alpha or beta) up regulated production of IL-3, IL-5, IL-6, and IL-9 as well as TNF, i.e., cytokines implicated in many inflammatory processes including those associated with allergies and helminthic infections. IL-1 did not induce significant cytokine release in the absence of ionomycin or IgE/Ag, suggesting that Ca dependent signaling was required. IL-1-mediated enhancement of cytokine expression was confirmed at the mRNA level by Northern blot and/or RT-PCR analysis. Our study reveals a role for IL-1 in the up-regulation of multiple mast cell-derived cytokines. Moreover, we identify mast cells as a novel source of IL 9. These results are of particular importance in the light of recent reports that strongly support a central role of IL-9 in allergic lung inflammation and in host defense against worm infections. PMID- 10820230 TI - Inhibition of lipopolysaccharide-induced signal transduction in endotoxin tolerized mouse macrophages: dysregulation of cytokine, chemokine, and toll-like receptor 2 and 4 gene expression. AB - In this study, the effect of in vitro endotoxin tolerance on LPS-induced mitogen activated protein kinase activation, transcription factor induction, and cytokine, chemokine, and Toll-like receptor (TLR) 2 and 4 gene expression, as well as the involvement of TNF and IL-1 signaling pathways in tolerance, were examined. Pretreatment of mouse macrophages with LPS inhibited phosphorylation of the extracellular signal-regulated kinases, c-Jun NH2-terminal kinases, and p38 kinase; degradation of I-kappaBalpha (inhibitory protein that dissociates from NF kappaB) and I-kappaBbeta; and activation of the transcription factors NF-kappaB and AP-1 in response to subsequent LPS stimulation. These changes were accompanied by suppression of LPS-induced expression of mRNA for GM-CSF, IFN gamma-inducible protein-10, KC, JE/monocyte chemoattractant protein-1, macrophage inflammatory protein-1beta, and macrophage-inflammatory protein-2, with concurrent inhibition of chemokine secretion. In contrast to control cells, endotoxin-tolerant macrophages exhibited an increased basal level of TLR2 mRNA, and failed to increase levels of TLR2 mRNA or to down-regulate TLR4 gene expression upon restimulation with LPS. As judged by transcription factor activation, LPS and IL-1 were found to induce a state of cross-tolerance against each other, while no such reciprocal effect was seen for LPS and TNF-alpha. In addition, macrophages from TNFR I/II double knockout mice were LPS tolerizable, and blocking of endogenous TNF-alpha with TNFR-Fc fusion protein did not affect the capacity of LPS to tolerize macrophages. These data extend our understanding of LPS-signaling mechanisms that are inhibited in endotoxin-tolerized macrophages and suggest that endotoxin tolerance might result from impaired expression and/or functions of common signaling intermediates involved in LPS and IL-1 signaling. PMID- 10820231 TI - Regulation of murine airway eosinophilia and Th2 cells by antigen-conjugated CpG oligodeoxynucleotides as a novel antigen-specific immunomodulator. AB - The characteristic features of bronchial asthma reflect the orchestrated activity of Th2 cells. Oligodeoxynucleotides containing CpG motifs (CpG) have recently been highlighted as an immunomodulator that biases toward a Th1-dominant phenotype. We have previously reported that intratracheal coadministration of CpG and allergen inhibited airway eosinophilia and hyperresponsiveness in a synergistic manner. To substantiate the synergism between CpG and Ag, we introduced a covalently linked conjugate between CpG and Ag and examined the efficacy on airway eosinophilia and Th2 cytokine production. We found that the conjugated form of CpG plus Ag was 100-fold more efficient in regulating airway eosinophilia than the unconjugated mixture. The inhibitory effects lasted for at least 2 mo. The inhibition of airway eosinophilia by the conjugate was Ag specific and associated with an improvement of the airway hyperresponsiveness and the unresponsiveness of the Ag-specific Th2 cells in the regional lymph nodes. The CpG-Ag conjugate was 100-fold more effective than the unconjugated mixture for inducing in vitro Th1 differentiation in an IL-12-dependent manner. Our data show that CpG conjugated to Ag can work as a novel Ag-specific immunomodulator and imply that inhalation of allergen-CpG conjugate could be a desensitization therapy for patients with bronchial asthma. PMID- 10820232 TI - A soluble lymphocyte activation gene-3 molecule used as a vaccine adjuvant elicits greater humoral and cellular immune responses to both particulate and soluble antigens. AB - The lymphocyte activation gene-3 (LAG-3) product is a MHC class II ligand that has been used in vivo to stimulate MHC class II+ APCs to increase tumor-specific immune responses. We investigated whether LAG-3 could also play an adjuvant role in vivo for the induction of humoral and CD4 or CD8 cell-mediated immune responses when immunizing mice with a particulate (hepatitis B surface Ag) or soluble (OVA) Ag. In both cases, coadministration of 1 microg of a soluble fusion protein between murine LAG-3 and the Fc fraction of a murine IgG2a mAb (mLAG-3Ig) as a vaccine adjuvant induced or increased CTL responses to the corresponding MHC class I-restricted peptide. In addition, splenocytes of mice vaccinated with either the particulate or soluble Ag plus mLAG-3Ig exhibited a significantly greater proliferative response than did splenocytes of mice immunized with Ag and a control Ig molecule. Similarly, these splenocytes had a greater Th1- but not Th2-type cytokine response. Finally, mice immunized with Ag plus mLAG-3Ig produced higher titers of Abs than mice immunized with Ag and a control Ig molecule. Thus, these data provide evidence of a novel means of improving the immunogenicity of subunit vaccines. PMID- 10820233 TI - Molecular basis for functional maturation of thymocytes: increase in c-fos translation with positive selection. AB - In the process of positive selection, immature CD4+8+ double positive (DP) thymocytes expressing TCR reactive to self-MHC by appropriate avidity develop into mature thymocytes. Positive selection involves not only down-regulation of either CD4 or CD8 but also acquisition of immunocompetent potential such as cell proliferation and cytokine production. To understand the molecular basis for such functional maturation during the positive selection process, we examined whether nonselected DP, selected DP, and CD4+8- single positive thymocytes possess the activation potential for signaling pathways from mitogen-activated protein kinases (extracellular signal-regulated kinase and c-Jun N-terminal kinase) to AP 1. In response to stimulation, a marked induction of c-Fos protein expression as well as cell proliferation is detected only in CD4+8- single positive cells but not in selected and nonselected DP cells, though mitogen-activated protein kinase activities and c-fos transcripts are equally induced. In the presence of proteasome inhibitors, c-Fos protein became detectable in selected DP cells but still not in nonselected DP cells, suggesting that DP cells receiving positive selection signals acquire the capacity to translate the c-fos gene, but it may not be sufficiently high to overcome the degradation of c-Fos protein. These data indicate that the translating ability of the c-fos gene is up-regulated in the thymic positive selection process, from nonselected DP to CD4+8- single positive cells through positively selected DP cells. The distinguished responsiveness to stimulation in thymocytes with and without positive selection may be a result in part of the distinct regulation of the c-fos gene at the translational level. PMID- 10820234 TI - Heterogeneity of tonsillar subepithelial B lymphocytes, the splenic marginal zone equivalents. AB - The VH4 genes expressed by both resting and in vivo-activated subepithelial (SE) B cells from human tonsils were studied. Resting SE B cells were subdivided according to the presence (IgDlow) or absence (IgM-only) of surface IgD. CD27 was abundant on activated SE B cells and low on resting IgM-only B cells. Resting IgDlow SE B cells could be subdivided into CD27low and CD27high cell fractions. Resting IgDlow SE B cells displayed VH4 genes with a substantial number of mutations (13/29 of the molecular clones were mutated), whereas 25/26 of the clones from resting IgM-only SE B cells were unmutated. Moreover, mutated VH4 genes were detected mainly within the CD27high cell fraction of the IgDlow SE B cells. Several identical unmutated VH4DJH sequences (11/32) were found in different molecular clones from resting IgM-only SE B cells, suggesting local cellular expansion. Both unmutated (14/25) and mutated (11/25) sequences were found in mu transcripts of activated SE B cells. Extensive mutation was observed in the gamma transcripts of activated SE B cells. Therefore, SE B cells are heterogeneous, being comprised of B cells with mutated Ig VH4 genes, that are Ag experienced B cells, and a subset of B cells with unmutated VH4 genes that are either virgin cells or cells driven by Ags that did not induce or select for V gene mutations. PMID- 10820235 TI - B cell responses to a peptide epitope. IX. The kinetics of antigen binding differentially regulates costimulatory capacity of activated B cells. AB - We explore the possible mechanism by which association rates of Ag with activated B cells influences the ability of the latter to selectively recruit Th subsets. Our system used cocultures of Ag-activated B and T cells, where the Ag was a synthetic peptide, G41CT3. Restimulation was with either peptide G41CT3 or its analogue, G28CT3. Peptide G28CT3 has been previously shown to display a higher on rate, relative to the homologous peptide G41CT3, of binding to G41CT3-activated B cells. This difference in on rates was eventually exerted at the level of IFN gamma, but not of IL-10, induction from T cells, with peptide G28CT3 proving more effective. However, various treatment regimens rendered peptide G41CT3 as potent as peptide G28CT3 at eliciting IFN-gamma responses from the above cultures. This included simultaneous treatment of B cells with peptide G41CT3 and the protein tyrosine kinase inhibitor tyrphostin. Alternatively, pretreatment of B cells with a peptide representing only the B cell epitope constituent of peptide G28CT3 (G28) was also equally effective. Subsequent experiments revealed that IFN-gamma production from activated T cells resulted from an engagement of CD28 by B7-1 on the B cell surface. Finally, the extent of cell surface B7-1 up-regulation on activated B cells was dependent on the on rate of Ag binding to the membrane bound Ig receptor. Thus, cumulative results suggest that the kinetics of Ag binding to activated B cells can differentially regulate intracellular signaling. This influences selective costimulatory molecule expression, with its consequent effects on relative Th subset activation. PMID- 10820236 TI - B cell responses to a peptide epitope. X. Epitope selection in a primary response is thermodynamically regulated. AB - We examine the etiological basis of hierarchical immunodominance of B cell epitopes on a multideterminant Ag. A model T-dependent immunogen, containing a single immunodominant B cell epitope, was used. The primary IgM response to this peptide included Abs directed against diverse determinants presented by the peptide. Interestingly, affinity of individual monomeric IgM Abs segregated around epitope recognized and was independent of their clonal origins. Furthermore, affinity of Abs directed against the immunodominant epitope were markedly higher than that of the alternate specificities. These studies suggested that the affinity of an epitope-specific primary response, and variations therein, may be determined by the chemical composition of epitope. This inference was supported by thermodynamic analyses of monomer IgM binding to Ag, which revealed that this interaction occurs at the expense of unfavorable entropy changes. Permissible binding required compensation by net enthalpic changes. Finally, the correlation between chemical composition of an epitope, the resultant affinity of the early primary humoral response, and its eventual influence on relative immunogenicity could be experimentally verified. This was achieved by examining the effect of various amino-terminal substitutions on immunogenicity of a, hitherto cryptic, amino-terminal determinant. Such experiments permitted delineation of a hierarchy of individual amino acid residues based on their influence; which correlated well with calculated Gibbs free energy changes that individual residue side chains were expected to contribute in a binding interaction. Thus, maturation of a T-dependent humoral response is initiated by a step that is under thermodynamic control. PMID- 10820237 TI - Determination of the relationship between T cell responsiveness and the number of MHC-peptide complexes using specific monoclonal antibodies. AB - We describe the generation of three mAbs that recognize the complex of the class II MHC molecule IEk bound to a peptide derived from the carboxyl terminus of moth cytochrome c (residues 95-103). Reactivities of these mAbs are sensitive to single alterations in the sequence of both helices of the MHC molecule and to the bound peptide. The epitopes of these reagents are distinct but overlap substantially. One of these mAbs specifically blocks lymphokine release by T cells responsive to this complex but not others. We have used another to examine how the number of complexes on an APC is related to its ability to stimulate T cells. We find that 200-400 complexes per cell are necessary and sufficient to induce a degree of stimulation, whereas maximum stimulation is achieved only if more than 5000 complexes are present. The analysis indicates that T cell activation is a stochastic process. PMID- 10820238 TI - Early block in maturation is associated with thymic involution in mammary tumor bearing mice. AB - We previously reported that mice implanted with mammary tumors show a progressive thymic involution that parallels the growth of the tumor. The involution is associated with a severe depletion of CD4+8+ thymocytes. We have investigated three possible mechanisms leading to this thymic atrophy: 1) increased apoptosis, 2) decreased proliferation, and 3) disruption of normal thymic maturation. The levels of thymic apoptosis were determined by propidium iodide and annexin V staining. A statistically significant, but minor, increase in thymic apoptosis in tumor-bearing mice was detected with propidium iodide and annexin V staining. The levels of proliferation were assessed by in vivo labeling with 5'-bromo-2' deoxyuridine (BrdU). The percentages of total thymocytes labeled 1 day following BrdU injection were similar in control and tumor-bearing mice. Moreover, the percentages of CD4-8- thymocytes that incorporated BrdU during a short term pulse (5 h) of BrdU were similar. Lastly, thymic maturation was evaluated by examining CD44 and CD25 expression among CD4-8- thymocytes. The percentage of CD44+ cells increased, while the percentage of CD25+ cells decreased among CD4-8- thymocytes from tumor-bearing vs control animals. Together, these findings suggest that the thymic hypocellularity seen in mammary tumor bearers is not due to a decreased level of proliferation, but, rather, to an arrest at an early stage of thymic differentiation along with a moderate increase in apoptosis. PMID- 10820239 TI - CD4+ T cell responses to self- and mutated p53 determinants during tumorigenesis in mice. AB - We analyzed CD4+ T helper responses to wild-type (wt) and mutated (mut) p53 protein in normal and tumor-bearing mice. In normal mice, we observed that although some self-p53 determinants induced negative selection of p53-reactive CD4+ T cells, other p53 determinants (cryptic) were immunogenic. Next, BALB/c mice were inoculated with J774 syngeneic tumor cell line expressing mut p53. BALB/c tumor-bearing mice mounted potent CD4+ T cell responses to two formerly cryptic peptides on self-p53. This response was characterized by massive production of IL-5, a Th2-type lymphokine. Interestingly, we found that T cell response was induced by different p53 peptides depending upon the stage of cancer. Mut p53 gene was shown to contain a single mutation resulting in the substitution of a tyrosine by a histidine at position 231 of the protein. Two peptides corresponding to wt and mutated sequences of this region were synthesized. Both peptides bound to the MHC class II-presenting molecule (Ed) with similar affinities. However, only mut p53.225-239 induced T cell responses in normal BALB/c mice, a result strongly suggesting that high-affinity wt p53.225 239 autoreactive T cells had been eliminated in these mice. Surprisingly, CD4+ T cell responses to both mut and wt p53.225-239 peptides were recorded in J774 tumor-bearing mice, a phenomenon attributed to the recruitment of low-avidity p53.225-239 self-reactive T cells. PMID- 10820240 TI - Mouse CD8+ CD122+ T cells with intermediate TCR increasing with age provide a source of early IFN-gamma production. AB - Although CD8+ IL-2Rbeta (CD122)+ T cells with intermediate TCR reportedly develop extrathymically, their functions still remain largely unknown. In the present study, we characterized the function of CD8+ CD122+ T cells with intermediate TCR of C57BL/6 mice. The proportion of CD8+ CD122+ T cells in splenocytes gradually increased with age, whereas CD8+ IL-2Rbeta-negative or -low (CD122-) T cells conversely decreased. The IFN-gamma production from splenocytes stimulated with immobilized anti-CD3 Ab in vitro increased with age, whereas the IL-4 production decreased. When sorted CD8+ CD122+ T cells were stimulated in vitro by the anti CD3 Ab, they promptly produced a much larger amount of IFN-gamma than did CD8+ CD122- T cells or CD4+ T cells, whereas only CD4+ T cells produced IL-4. The depletion of CD8+ CD122+ T cells from whole splenocytes greatly decreased the CD3 stimulated IFN-gamma production and increased the IL-4 production, whereas the addition of sorted CD8+ CD122+ T cells to CD8+ CD122+ T cell-depleted splenocytes restored the IFN-gamma production and partially decreased IL-4 production. It is of interest that CD8+ CD122+ T cells stimulated CD4+ T cells to produce IFN gamma. The CD3-stimulated IFN-gamma production from each T cell subset was augmented by macrophages. Furthermore, CD3-stimulated CD8+ CD122+ T cells produced an even greater amount of IFN-gamma than did liver NK1.1+ T cells and also showed antitumor cytotoxicity. These results show that CD8+ CD122+ T cells may thus be an important source of early IFN-gamma production and are suggested to be involved in the immunological changes with aging. PMID- 10820241 TI - Constitutive expression of a chimeric receptor that delivers IL-2/IL-15 signals allows antigen-independent proliferation of CD8+CD44high but not other T cells. AB - We have prepared transgenic mice whose T cells constitutively express a chimeric receptor combining extracellular human IL-4R and intracellular IL-2Rbeta segments. This receptor can transmit IL-2/IL-15-like signals in response to human, but not mouse, IL-4. We used these animals to explore to what extent functional IL-2R/IL-15R expression controls the capacity of T cells to proliferate in response to IL-2/IL-15-like signals. After activation with Con A, naive transgenic CD8+ and CD4+ T cells respond to human IL-4 as well as to IL-2. Without prior activation, they failed to proliferate in response to human IL-4, although human IL-4 did prolong their survival. Thus, IL-2-induced proliferation of activated T cells requires at least one other Ag-induced change apart from the induction of a functional IL-2R. However, a fraction of CD8+CD44high T cells proliferate in human IL-4 without antigenic stimulation or syngeneic feeder cells. In contrast, CD4+CD44high T cells are not constitutively responsive to human IL-4. We conclude that although all transgenic T cells express a functional chimeric receptor, only some CD8+CD44high T cells contain all molecules required for entry into the cell cycle in response to human IL-4 or IL-15. PMID- 10820242 TI - Down-regulation of the orphan nuclear receptor ROR gamma t is essential for T lymphocyte maturation. AB - Thymocyte development is a tightly regulated process. CD4+CD8+ double-positive (DP) immature thymocytes exhibit distinct phenotypic features from mature T cells; they express only 10% of surface TCR that are found on mature T cells and do not proliferate and produce IL-2 in response to stimulation. In this report we show that transgenic expression of the orphan nuclear receptor ROR gamma t in mature T cells down-regulates their surface TCR expression. The ROR gamma t transgene inhibits IL-2 production by mature T cells, and this inhibition may be partially due to the inhibitory effect of ROR gamma t on c-Rel transcription. Furthermore, ectopic expression of ROR gamma t inhibits the proliferation of mature and immature T cells. These results, together with its predominant expression in DP thymocytes, suggest that ROR gamma t controls these distinct phenotypic features of DP thymocytes. Our data suggest that down-regulation of ROR gamma t expression in thymocytes is essential for their maturation. PMID- 10820243 TI - Ligand-specific selection of MHC class II-restricted thymocytes in fetal thymic organ culture. AB - Positive and negative selection of thymocytes is determined by the specificity of the TCR and signaling through its associated molecules. We have studied selection of thymocytes bearing a MHC class II-restricted TCR using fetal thymic organ culture. This system allows the addition of peptides to the already diverse panoply of endogenous peptide ligands and is useful for analyzing ligand-specific negative selection of CD4 single positive (CD4SP) thymocytes. The data reveal that the ability of a given ligand to mediate negative selection is related to its dissociation rate from the TCR. We find that negative selection is very sensitive, and only the weakest ligand that we can identify fails to induce negative selection. None of the numerous peptides tested were able to induce an increase in CD4SP thymocytes. In addition, the ligands that induce negative selection of CD4SP thymocytes also cause an increase in numbers of CD8SP thymocytes bearing high levels of the class II-restricted TCR. Although these cells have a cell surface phenotype consistent with positive selection, they most likely represent cells in the process of negative selection. Further analysis reveals that these cells are not induced by these ligands in intact adult animals and that their induction is probably only revealed in the organ culture system. PMID- 10820244 TI - CpG oligonucleotides are potent adjuvants for the activation of autoreactive encephalitogenic T cells in vivo. AB - The mechanism of action of microbial adjuvants in promoting the differentiation of autoimmune effector cells remains to be elucidated. We demonstrate that CpG containing oligodeoxynucleotides (ODN) can completely substitute for heat-killed mycobacteria in the priming of encephalitogenic myelin-reactive T cells in vivo. The adjuvanticity of the CpG ODN was secondary to their direct ability to induce IL-12 or to act synergistically with endogenous IL-12 to promote Th1 differentiation and encephalitogenicity. T cells primed in the absence of CpG with Ag and IFA alone appeared to be in a transitional state and had not undergone differentiation along a conventional Th pathway. Unlike Th2 cells, they expressed low levels of the IL-12R beta 2 subunit and retained the ability to differentiate into encephalitogenic effectors when reactivated in vitro under Th1 polarizing conditions. These results support the use of CpG ODN as adjuvants but also suggest that they could potentially trigger autoimmune disease in a susceptible individual. PMID- 10820245 TI - Cooperation among Stat1, glucocorticoid receptor, and PU.1 in transcriptional activation of the high-affinity Fc gamma receptor I in monocytes. AB - IFN-gamma and glucocorticoids regulate inflammatory and immune responses through Stat1 and glucocorticoid receptor (GR) transcription factors, respectively. The biological responses to these polypeptides are determined by integration of various signaling pathways in a cell-type and promoter-dependent manner. In this study we have characterized the molecular basis for the functional cooperation between IFN-gamma and dexamethasone (Dex) in the induction of the high-affinity Fc gamma receptor I (Fc gamma RI) in monocytes. Dex did not affect IFN-gamma induced Stat1 DNA binding activity or induce novel DNA-binding complexes to the Fc gamma RI promoter. By using cell systems lacking functional GR or Stat1, we showed that GR stimulated Stat1-dependent transcription in a ligand-dependent manner, while Stat1 did not influence GR-dependent transcription. The cooperation required phosphorylation of Tyr701, DNA binding, and the trans-activation domain of Stat1, but did not involve Ser727 phosphorylation of Stat1 or physical interaction between GR and Stat1. The costimulatory effect of Dex was not dependent on a consensus glucocorticoid response element in the Stat1-responsive promoters, but required the DNA-binding and trans-activation functions of GR, and Dex-induced protein synthesis. GR activated the natural Fc gamma RI promoter construct, and this response required both Stat1 and the Ets family transcription factor PU.1. Previously, physical association between GR and Stat5 has been shown to enhance Stat5-dependent and suppress GR-dependent transcription. The results shown here demonstrate a distinct, indirect mechanism of cross-modulation between cytokine and steroid receptor signaling that integrates Stat1 and GR pathways with cell type-specific PU.1 transcription factor in the regulation of Fc gamma RI gene transcription. PMID- 10820246 TI - Antigenic epitopes regulate the phenotype of CD8+ CTL primed by exogenous antigens. AB - We previously reported that insulin-specific, MHC class I-restricted CTL precursors can be primed by injecting C57BL/6 mice with bovine insulin in CFA. These bovine insulin-primed CTL displayed a type 0 CTL phenotype, producing IL-4, IL-5, IL-10, low levels of IFN-gamma, but no TNF-alpha. By contrast, CTL generated from C57BL/6 mice primed with OVA in CFA produced IFN-gamma and TNF alpha but no IL-4, IL-5, or IL-10 and therefore were classified as type 1 CTL. Although CD4+ T cell subsets have been compared extensively in the literature, CTL subsets are less well characterized. Here, the phenotype, function, and requirements for the in vivo activation of type 1 and type 0 CTL cells were studied. Although both types of CTL express many of the same cell-surface Ags, OVA-specific CTL but not bovine insulin-primed CTL expressed CT-1, a carbohydrate epitope of CD45, and bovine insulin-primed CTL but not OVA-specific CTL expressed Fas constitutively. Priming of CTL was abrogated by depletion of phagocytic cells but not CD4+ T cells, whereas depletion of CD4+ T cells but not phagocytic cells inhibited Ab responses in the same mice. Neither endogenous IL-4 nor the dose of priming Ag altered the CTL phenotypes, but the antigenic peptides of OVA and bovine insulin were key to determining the differentiation of either type 1 or type 0 CTL. To our knowledge, this is the first time that antigenic epitopes have been demonstrated to influence the phenotype of Ag-specific CTL responses. These results may be relevant to the development of peptide vaccines in which a particular type of CTL response is desired. PMID- 10820247 TI - IL-4 promotes the migration of circulating B cells to the spleen and increases splenic B cell survival. AB - We report that IL-4 causes a redistribution of B cells and modestly increases B cell life span. Intravenous injection of a long-acting formulation of IL-4 induces increases in both spleen cell number and the percentage of splenic B cells. These effects are observed within 1 day of IL-4 administration and plateau after approximately 3 days if IL-4 treatment is continued. The increase in splenic B cell number is IL-4 dose dependent, CD4+ T cell independent, FcgammaRII/FcgammaRIII independent, and Stat6 independent. Decreases in the number of B cells in the blood and the percentage of mature B cells in the bone marrow, concomitant with the increase in splenic B cell number, suggest that redistribution of circulating B cells to the spleen is partially responsible for IL-4 induction of splenic B cell hyperplasia. Considerable reduction in the effect of 5 days of IL-4 treatment on splenic B cell number when B lymphopoiesis is blocked with anti-IL-7 mAb suggests that generation of new B cells is also involved in IL-4-induced splenic B cell hyperplasia. 5-Bromo-2'-deoxyuridine labeling experiments demonstrate that IL-4 modestly prolongs the life span of newly generated splenic B cells, and experiments that measure B cell HSA (CD24) expression as an indicator of B cell age suggest that IL-4 may also prolong the life span of mature splenic B cells. Thus, IL-4 increases splenic B cell number through two Stat6-independent effects: increased net migration of circulating B cells to the spleen and increased B cell life span. Both effects may promote Ab responses to a systemic Ag challenge. PMID- 10820248 TI - Regulation of extrathymic T cell development and turnover by oncostatin M. AB - Chronic exposure to oncostatin M (OM) has been shown to stimulate extrathymic T cell development. The present work shows that in OM transgenic mice, 1) massive extrathymic T cell development takes place exclusively the lymph nodes (LNs) and not in the bone marrow, liver, intestines, or spleen; and 2) LNs are the sole site where the size of the mature CD4+ and CD8+ T cell pool is increased (6- to 7 fold). Moreover, when injected into OM transgenic mice, both transgenic and nontransgenic CD4+ and CD8+ T cells preferentially migrated to the LNs rather than the spleen. Studies of athymic recipients of fetal liver grafts showed that lymphopoietic pathway modulated by OM was truly thymus independent, and that nontransgenic progenitors could generate extrathymic CD4+CD8+ cells as well as mature T cells under the paracrine influence of OM. The progeny of the thymic independent differentiation pathway regulated by OM was polyclonal in terms of Vbeta usage, exhibited a phenotype associated with previous TCR ligation, and displayed a rapid turnover rate (5-bromo-2'-deoxyuridine pulse-chase assays). This work suggests that chronic exposure to OM 1) discloses a unique ability of LNs to sustain extrathymic T cell development, and 2) increases the number and/or function of LN niches able to support seeding of recirculating mature T cells. Regulation of the lymphopoietic pathway discovered in OM transgenic mice could be of therapeutic interest for individuals with thymic hypoplasia or deficient peripheral T cell niches. PMID- 10820249 TI - Gestational age-dependent expression of IL-10 and its receptor in human placental tissues and isolated cytotrophoblasts. AB - Control of antifetal immune responses is thought to be regulated locally by the placenta. Because the physiologic programming of the placenta across gestation is likely to influence the local immunity, we hypothesize that a potent anti inflammatory cytokine such as IL-10 may be produced in a gestational age dependent manner. In the present study, we examined the expression of IL-10 and its receptor in placental explants or freshly isolated cytotrophoblasts from different gestational ages and compared it with the expression profiles of other cytokines. First and second trimester placental tissues from normal pregnancies predominantly expressed IL-10, whereas the levels of IL-2, IL-4, and IFN-gamma were mostly below detection throughout pregnancy. The expression of IL-10, but not its receptor, diminished significantly in term placental tissues collected "before" the onset of labor and did not change appreciably "after" labor. On the other hand, TNF-alpha and IL-1beta were significantly up-regulated in response to labor-associated conditions. IL-10 expression was transcriptionally attenuated at term as observed in cytotrophoblasts. In contrast to the placental cytokine milieu, autologous PBMCs, when activated with PHA, secreted significant amounts of IL-2, IL-4, IL-10, and IFN-gamma, albeit with a statistically significantly enhanced IL-10 production in first trimester compared with age-matched nonpregnant women. These data suggest that IL-10 is expressed in the placenta in a gestational age-dependent manner and that its down-regulation at term may be an important mechanism underlying the subtle changes associated with parturition. PMID- 10820251 TI - Regulatory cells potentiate the efficacy of IL-4 gene transfer by up-regulating Th2-dependent expression of protective molecules in the infectious tolerance pathway in transplant recipients. AB - We have previously shown that the tolerant state in allograft recipients can be maintained and perpetuated by an "infectious" T cell-dependent regulatory mechanism. Hence, 1) treatment of LEW rats with RIB-5/2, a CD4 nondepleting mAb, produces indefinite survival of LBNF1 cardiac allografts; 2) donor-specific tolerance can be then transferred by spleen cells into new cohorts of test allograft recipients; and 3) putative regulatory CD4+ Th2-like cells are instrumental in this tolerance model. We now report on studies aimed at exposing mechanisms underlying the infectious tolerance pathway, with emphasis on the interactions between intragraft adenovirus-IL-4 gene transfer and systemic infusion of regulatory cells from tolerant hosts. Unlike individual treatment regimens, adjunctive therapy with adenovirus-IL-4 and suboptimal doses of regulatory spleen cells was strongly synergistic and extended donor-type test cardiac allograft survival to about 2 mo. RT-PCR-based expression of intragraft mRNA coding for IL-2 and IFN-gamma remained depressed, whereas that of IL-4 and IL-10 reciprocally increased selectively in the combined treatment group, data supported by ELISA studies. In parallel, only adjunctive treatment triggered intragraft induction of molecules with anti-oxidant (HO-1) and anti-apoptotic (Bcl-xL/Bag-1) but not with pro-apoptotic (CPP-32) functions, both in the early and late posttransplant phases. Hence, systemic infusion of regulatory cells potentiates the effects of local adenovirus-IL-4 gene transfer in transplant recipients. Th2-driven up-regulation of protective molecule programs at the graft site, such as of anti-oxidant HO-1 and/or anti-apoptotic Bcl-xL and Bag-1, may contribute, at least in part, to the maintenance of the infectious tolerance pathway in transplant recipients. PMID- 10820250 TI - Identification of murine germinal center B cell subsets defined by the expression of surface isotypes and differentiation antigens. AB - Germinal centers (GCs) are inducible lymphoid microenvironments that support the generation of memory B cells, affinity maturation, and isotype switching. Previously, phenotypic transitions following in vivo B cell activation have been exploited to discriminate GC from non-GC B cells in the mouse and to delineate as many as seven distinct human peripheral B cell subsets. To better understand the differentiative processes occurring within murine GCs, we sought to identify subpopulations of GC B cells corresponding to discrete stages of GC B cell ontogeny. We performed multiparameter flow-cytometric analyses of GC B cells at consecutive time points following immunization of BALB/c mice with SRBC. We resolved the murine GC compartment into subsets based on the differential expression of activation markers, surface Ig isotypes, and differentiation Ags. Class-switched and nonswitched GC B cells emerged contemporaneously, and their relative frequencies remained nearly constant throughout the GC reaction, perhaps reflecting the establishment of a steady state. A significant percentage of the nonswitched B cells with a GC phenotype exhibited surface markers associated with naive B cells, including CD23, surface IgD, and high levels of CD38 consistent with either prolonged recruitment into the GC reaction or protracted expression of these markers during differentiation within the GC. Expression of the activation marker BLA-1 was dynamic over time, with all GC B cells being positive early after immunization, followed by progressive loss as the GC reaction matured into the second and third week. Implications of these results concerning GC evolution are discussed. PMID- 10820252 TI - Differential effect of cytokine treatment on Fc alpha receptor I- and Fc gamma receptor I-mediated tumor cytotoxicity by monocyte-derived macrophages. AB - Macrophages represent an important effector cell for Ab-mediated tumor therapy. Previous studies have documented that cytokines can influence Fc receptor (FcR) expression and function. In this study we examined the tumoricidal activities of the type I receptors for IgG (Fc gamma RI) and the IgA FcR (Fc alpha RI) on monocyte-derived macrophages (MDM) cultured in the presence of IFN-gamma, M-CSF, or GM-CSF. Bispecific Abs were used to target a Her2/neu breast carcinoma cell line, SKBR-3, to Fc alpha RI or Fc gamma RI on MDM. Although Fc alpha RI and Fc gamma RI share a common signaling pathway contingent on association with the gamma-chain (FcR gamma subunit), a marked difference in their efficiency in mediating tumoricidal functions was seen in response to specific cytokines. M-CSF and GM-CSF-treated MDM mediated efficient phagocytosis of SKBR-3 cells by Fc alpha RI and Fc gamma RI; however, IFN-gamma-treated MDM phagocytosed tumor cells only with the Fc gamma RI-directed bispecific Abs. Similarly, IFN-gamma-cultured MDM lysed tumor cells more efficiently via Fc gamma RI then by Fc alpha RI as measured in Ab-dependent cellular cytotoxicity assays. Conversely, GM-CSF-treated MDM mediated more efficient lysis of tumor cells via Fc alpha RI than Fc gamma RI, while M-CSF-cultured MDM were relatively less efficient in mediating Ab dependent cellular cytotoxicity through either receptor. With the exception of IFN-gamma-mediated enhancement of Fc gamma RI expression and Fc gamma RI gamma chain complexes, the regulation of Fc gamma RI- or Fc alpha RI-mediated activity occurred without significant change in either receptor expression or total complexes with gamma subunit. These data suggest that the efficiency of Ab mediated tumor therapy, which depends on FcR effector cell functions, may be modified by the use of specific cytokines. PMID- 10820253 TI - Hoxa3 and pax1 transcription factors regulate the ability of fetal thymic epithelial cells to promote thymocyte development. AB - Thymocyte maturation into T cells depends on interactions between thymocytes and thymic epithelial cells. In this study, we show that mutations in two transcription factors, Hoxa3 and Pax1, act synergistically to cause defective thymic epithelial cell development, resulting in thymic ectopia and hypoplasia. Hoxa3+/-Pax1-/- compound mutant mice exhibited more severe thymus defects than Pax1-/- single mutants. Fetal liver adoptive transfer experiments revealed that the defect resided in radio-resistant stromal cells and not in hematopoietic cells. Compound mutants have fewer MHC class II+ epithelial cells, and the level of MHC expression detected was lower. Thymic epithelial cells in these mutants have reduced ability to promote thymocyte development, causing a specific block in thymocyte maturation at an early stage that resulted in a dramatic reduction in the number of CD4+8+ thymocytes. This phenotype was accompanied by increased apoptosis of CD4+8+ thymocytes and their immediate precursors, CD44-25-(CD3-4-8-) cells. Our results identify a transcriptional regulatory pathway required for thymic epithelial cell development and define multiple roles for epithelial cell regulation of thymocyte maturation at the CD4-8- to CD4+8+ transition. PMID- 10820254 TI - FTY720 immunosuppression impairs effector T cell peripheral homing without affecting induction, expansion, and memory. AB - FTY720 (2-amino-2-(2-[4-octylphenyl]ethyl)-1,3-propanediol hydrochloride) prolongs survival of solid organ allografts in animal models. Mechanisms of FTY720 immunomodulation were studied in mice infected with lymphocytic choriomeningitis virus (LCMV) to assess T cell responses or with vesicular stomatitis virus to evaluate Ab responses. Oral FTY720 (0.3 mg/kg/day) did not affect LCMV replication and specific CTL and B cells were induced and expanded normally. Moreover, the anti-viral humoral immune responses were normal. However, FTY720 treatment showed first a shift of overall distribution of CTL from the spleen to peripheral lymph nodes and lymphocytopenia was observed. This effect was reversible within 7-21 days. Together with unimpaired T and B cell memory after FTY720 treatment, this finding rendered enhancement of lymphocyte apoptosis by FTY720 in vivo unlikely. Secondly, the delayed-type hypersensitivity reaction to a viral MHC class I-presented peptide was markedly reduced by FTY720. These results were supported by impaired circulation of LCMV specific TCR transgenic effector lymphocytes in the peripheral blood and reduced numbers of tissue infiltrating CTL in response to delayed-type hypersensitivity reaction. Thirdly, in a CD8+ T cell-mediated diabetes model in a transgenic mouse expressing the LCMV glycoprotein in the islets of the pancreas, FTY720 delayed or prevented disease by reducing islet-infiltrating CTL. Thus, FTY720 effectively reduced recirculation of CD8+ effector T cells and their recruitment to peripheral lesions without affecting the induction and expansion of immune responses in secondary lymphoid organs. These properties may offer the potential to treat ongoing organ-specific T cell-mediated immunopathologic disease. PMID- 10820255 TI - Revisiting tolerance induced by autoantigen in incomplete Freund's adjuvant. AB - Injection of autoantigens in IFA has been one of the most effective ways of preventing experimental, T cell-mediated, autoimmune disease in mice. The mechanism that underlies this protection has, however, remained controversial, with clonal deletion, induction of suppressor cells or of type 2 immunity being implicated at one time or another. Using high resolution enzyme-linked immunospot (ELISPOT) analysis, we have revisited this paradigm. As models of autoimmunity against sequestered and readily accessible autoantigens, we studied experimental allergic encephalomyelitis, induced by myelin oligodendrocyte glycoprotein, proteolipid protein, myelin basic protein, and renal tubular Ag-induced interstitial nephritis. We showed that the injection of each of these Ags in IFA was immunogenic and CD4 memory cells producing IL-2, IL-4, and IL-5, but essentially no IFN-gamma. IgG1, but not IgG2a, autoantibodies were produced. The engaged T cells were not classic Th2 cells in that IL-4 and IL-5 were produced by different cells. The IFA-induced violation of self tolerance, including the deposition of specific autoantibodies in the respective target organs, occurred in the absence of detectable pathology. Exhaustion of the pool of naive precursor cells was shown to be one mechanism of the IFA-induced tolerance. In addition, while the IFA-primed T cells acted as suppressor cells, in that they adoptively transferred disease protection, they did not interfere with the emergence of a type 1 T cell response in the adoptive host. Both active and passive tolerance mechanisms, therefore, contribute to autoantigen:IFA-induced protection from autoimmune disease. PMID- 10820256 TI - Size estimate of the alpha beta TCR repertoire of naive mouse splenocytes. AB - The diversity of the T cell repertoire of mature T splenocytes is generated, in the thymus, by pairing of alpha and beta variable domains of the alpha beta TCR and by the rearrangements of various gene segments encoding these domains. In the periphery, it results from competition between various T cell subpopulations including recent thymic migrants and long-lived T cells. Quantitative data on the actual size of the T cell repertoire are lacking. Using PCR methods and extensive sequencing, we have measured for the first time the size of the TCR-alpha beta repertoire of naive mouse T splenocytes. There are 5-8 x 105 different nucleotide sequences of BV chains in the whole spleen of young adult mice. We have also determined the size of the BV repertoire in a subpopulation of AV2+ T splenocytes, which allows us to provide a minimum estimate of the alpha beta repertoire. We find that the mouse spleen harbors about 2 x 106 clones of about 10 cells each. This figure, although orders of magnitude smaller than the maximum theoretical diversity (estimated up to 1015), is still large enough to maintain a high functional diversity. PMID- 10820257 TI - Molecular basis for recognition of an arthritic peptide and a foreign epitope on distinct MHC molecules by a single TCR. AB - KRN TCR transgenic T cells recognize two self-MHC molecules: a foreign peptide, bovine RNase 42-56, on I-Ak and an autoantigen, glucose-6-phosphate isomerase 282 294, on I-Ag7. Because the latter recognition event initiates a disease closely resembling human rheumatoid arthritis, we investigated the structural basis of this pathogenic TCR's dual specificity. While peptide recognition is altered to a minor degree between the MHC molecules, we show that the receptor's cross reactivity critically depends upon a TCR contact residue completely conserved in the foreign and self peptides. Further, the altered recognition of peptide derives from discrete differences on the MHC recognition surfaces and not the disparate binding grooves. This work provides a detailed structural comparison of an autoreactive TCR's interactions with naturally occurring peptides on distinct MHC molecules. The capacity to interact with multiple self-MHCs in this manner increases the number of potentially pathogenic self-interactions available to a T cell. PMID- 10820258 TI - KIR2DL5, a novel killer-cell receptor with a D0-D2 configuration of Ig-like domains. AB - Four novel killer-cell Ig-like receptor (KIR) genes were discovered by analysis of genomic DNA from a human donor. One gene, KIR2DL5, is expressed by subpopulations of NK cells and T cells, whereas expression of the other three genes could not be detected. KIR2DL5 has two extracellular Ig-like domains of the D0 and D2 type, a structural configuration that was previously unique to KIR2DL4. Although having a similar structure overall, the KIR2DL4 and KIR2DL5 receptors have distinctive amino acid sequences in the ligand-binding extracellular domains and differ in the transmembrane and cytoplasmic motifs that determine signal transduction. Whereas the KIR2DL4 gene is present on all KIR haplotypes and is expressed by all human NK cells, the KIR2DL5 gene is restricted to the "B" subset of KIR haplotypes and is clonally expressed by NK cells within an individual. Chimpanzee genes for KIR2DL4 and KIR2DL5 have been defined and are very similar in sequence to their human orthologs. The donor in whom KIR2DL5 was first detected bears two variants of it that differ by five nucleotide substitutions in the coding region. Although the substitutions are not predicted to affect gene expression, transcription of only one of the two KIR2DL5 variants could be detected. PMID- 10820259 TI - GRID: a novel Grb-2-related adapter protein that interacts with the activated T cell costimulatory receptor CD28. AB - Adapter proteins such as Grb2 play a central role in the formation of signaling complexes through their association with multiple protein binding partners. These interactions are mediated by specialized domains such as the well-characterized Src homology SH2 and SH3 motifs. Using yeast three-hybrid technology, we have identified a novel adapter protein, expressed predominantly in T lymphocytes, that associates with the activated form of the costimulatory receptor, CD28. The protein is a member of the Grb2 family of adapter proteins and contains an SH3 SH2-SH3 domain structure. A unique glutamine/proline-rich domain (insert domain) of unknown function is situated between the SH2 and N-terminal SH3 domains. We term this protein GRID for Grb2-related protein with insert domain. GRID coimmunoprecipitates with CD28 from Jurkat cell lysates following activation of CD28. Using mutants of CD28 and GRID, we demonstrate that interaction between the proteins is dependent on phosphorylation of CD28 at tyrosine 173 and integrity of the GRID SH2 domain, although there are also subsidiary stabilizing contacts between the PXXP motifs of CD28 and the GRID C-terminal SH3 domain. In addition to CD28, GRID interacts with a number of other T cell signaling proteins, including SLP-76 (SH2 domain-containing leukocyte protein of 76 kDa), p62dok, and RACK-1 (receptor for activated protein kinase C-1). These findings suggest that GRID functions as an adapter protein in the CD28-mediated costimulatory pathway in T cells. PMID- 10820260 TI - Vesnarinone suppresses TNF-induced activation of NF-kappa B, c-Jun kinase, and apoptosis. AB - Vesnarinone, a synthetic quinolinone derivative used in the treatment of cardiac failure, exhibits immunomodulatory, anti-inflammatory, and cell growth regulatory properties. The mechanisms underlying these properties are not understood, but due to the critical role of nuclear transcription factor NF-kappa B in these responses, we hypothesized that vesnarinone must modulate NF-kappa B activation. We investigated the effect of vesnarinone on NF-kappa B activation induced by inflammatory agents. Vesnarinone blocked TNF-induced activation of NF-kappa B in a concentration- and time-dependent manner. This effect was mediated through inhibition of phosphorylation and degradation of I kappa B alpha, an inhibitor of NF-kappa B. The effects of vesnarinone were not cell type specific, as it blocked TNF-induced NF-kappa B activation in a variety of cells. NF-kappa B-dependent reporter gene transcription activated by TNF was also suppressed by vesnarinone. The TNF-induced NF-kappa B activation cascade involving TNF receptor 1-TNF receptor associated death domain-TNF receptor associated factor 2 NF-kappa B inducing kinase-IKK was interrupted at the TNF receptor associated factor 2 and NF-kappa B-inducing kinase sites by vesnarinone, thus suppressing NF-kappa B reporter gene expression. Vesnarinone also blocked NF-kappa B activation induced by several other inflammatory agents, inhibited the TNF-induced activation of transcription factor AP-1, and suppressed the TNF-induced activation of c-Jun N terminal kinase and mitogen-activated protein kinase kinase. TNF-induced cytotoxicity, caspase activation, and lipid peroxidation were also abolished by vesnarinone. Overall, our results indicate that vesnarinone inhibits activation of NF-kappa B and AP-1 and their associated kinases. This may provide a molecular basis for vesnarinone's ability to suppress inflammation, immunomodulation, and growth regulation. PMID- 10820261 TI - Definition of minimal domains of interaction within the recombination-activating genes 1 and 2 recombinase complex. AB - During V(D)J recombination, recognition and cleavage of the recombination signal sequences (RSSs) requires the coordinated action of the recombination-activating genes 1 and 2 (RAG1/RAG2) recombinase complex. In this report, we use deletion mapping and site-directed mutagenesis to determine the minimal domains critical for interaction between RAG1 and RAG2. We define the active core of RAG2 required for RSS cleavage as aa 1-371 and demonstrate that the C-terminal 57 aa of this core provide a dominant surface for RAG1 interaction. This region corresponds to the last of six predicted kelch repeat motifs that have been proposed by sequence analysis to fold RAG2 into a six-bladed beta-propeller structure. Residue W317 within this sixth repeat is shown to be critical for mediating contact with RAG1 and concurrently for stabilizing binding and directing cleavage of the RSS. We also show that zinc finger B (aa 727-750) of RAG1 provides a dominant interaction domain for recruiting RAG2. In all, the data support a model of RAG2 as a multimodular protein that utilizes one of its six faces for establishing productive contacts with RAG1. PMID- 10820262 TI - IFN regulatory factor-1-mediated transcriptional activation of mouse STAT-induced STAT inhibitor-1 gene promoter by IFN-gamma. AB - STAT-induced STAT inhibitor-1 (SSI-1), also referred to as suppressor of cytokine signaling-1 and JAK-binding protein, is a member of a new family, the members of which are negative regulators of cytokine signals. SSI-1 is induced by various cytokines; however, the transcriptional mechanism of the SSI-1 gene is not fully understood. Here, we showed that transcription of the mouse SSI-1 gene was initiated from six adjoining sites accompanying three GC boxes and a single GC box-like element near them, but not from the TATA box or an initiator sequence. We also showed that IFN-gamma induced SSI-1 mRNA more strongly than IL-6 in NIH 3T3 fibroblasts and that this IFN-gamma effect was mediated by Stat1. To determine the signal pathway downstream of Stat1, transcriptional activities of several mutant promoters were examined. The region mediating stimulatory effect of IFN-gamma to the gene transcription was localized to the -88/-60 region containing three tandem GAAA units, named variant IFN-gamma-responsive element (VIRE), while four IFN-gamma activation site (GAS)-like elements located far upstream were not related to the IFN-gamma response. Gel-shift assays revealed that IFN-gamma induced IFN regulatory factor-1 (IRF-1) binding to VIRE, but not that of IRF-2 or three components of ISGF3. Furthermore, forced expression of IRF 1 mimicked and that of IRF-2 inhibited the stimulatory effect of IFN-gamma on SSI 1 gene transcription. Finally, mouse embryonal fibroblasts lacking IRF-1 showed impaired SSI-1 mRNA induction by IFN-gamma. These results demonstrated that IRF 1, which is induced by activation of Stat1, mediated transcriptional activation of the SSI-1 gene by IFN-gamma via VIRE. PMID- 10820263 TI - Structure of the major peanut allergen Ara h 1 may protect IgE-binding epitopes from degradation. AB - In the past decade, there has been an increase in allergic reactions to peanut proteins, sometimes resulting in fatal anaphylaxis. The development of improved methods for diagnosis and treatment of peanut allergies requires a better understanding of the structure of the allergens. Ara h 1, a major peanut allergen belonging to the vicilin family of seed storage proteins, is recognized by serum IgE from >90% of peanut-allergic patients. In this communication, Ara h 1 was shown to form a highly stable homotrimer. Hydrophobic interactions were determined to be the main molecular force holding monomers together. A molecular model of the Ara h 1 trimer was constructed to view the stabilizing hydrophobic residues in the three dimensional structure. Hydrophobic amino acids that contribute to trimer formation are at the distal ends of the three dimensional structure where monomer-monomer contacts occur. Coincidentally, the majority of the IgE-binding epitopes are also located in this region, suggesting that they may be protected from digestion by the monomer-monomer contacts. On incubation of Ara h 1 with digestive enzymes, various protease-resistant fragments containing IgE-binding sites were identified. The highly stable nature of the Ara h 1 trimer, the presence of digestion resistant fragments, and the strategic location of the IgE-binding epitopes indicate that the quaternary structure of a protein may play a significant role in overall allergenicity. PMID- 10820264 TI - Soluble NSF attachment protein receptors (SNAREs) in RBL-2H3 mast cells: functional role of syntaxin 4 in exocytosis and identification of a vesicle associated membrane protein 8-containing secretory compartment. AB - Mast cells upon stimulation through high affinity IgE receptors massively release inflammatory mediators by the fusion of specialized secretory granules (related to lysosomes) with the plasma membrane. Using the RBL-2H3 rat mast cell line, we investigated whether granule secretion involves components of the soluble N ethylmaleimide-sensitive factor attachment protein receptor (SNARE) machinery. Several isoforms of each family of SNARE proteins were expressed. Among those, synaptosome-associated protein of 23 kDa (SNAP23) was central in SNARE complex formation. Within the syntaxin family, syntaxin 4 interacted with SNAP23 and all vesicle-associated membrane proteins (VAMPs) examined, except tetanus neurotoxin insensitive VAMP (TI-VAMP). Overexpression of syntaxin 4, but not of syntaxin 2 nor syntaxin 3, caused inhibition of FcepsilonRI-dependent exocytosis. Four VAMP proteins, i.e., VAMP2, cellubrevin, TI-VAMP, and VAMP8, were present on intracellular membrane structures, with VAMP8 residing mainly on mediator containing secretory granules. We suggest that syntaxin 4, SNAP23, and VAMP8 may be involved in regulation of mast cell exocytosis. Furthermore, these results are the first demonstration that the nonneuronal VAMP8 isoform, originally localized on early endosomes, is present in a regulated secretory compartment. PMID- 10820265 TI - IL-12p70 production by Leishmania major-harboring human dendritic cells is a CD40/CD40 ligand-dependent process. AB - Leishmaniasis, a vector-borne parasitic disease, is transmitted during a sandfly blood meal as the parasite is delivered into the dermis. The parasite displays a unique immune evasion mechanism: prevention of IL-12 production within its host cell, the macrophage (i.e., where it differentiates and multiplies). Given the close proximity of skin dendritic cells (DC) to the site of parasite delivery, their critical role in initiating immune responses and the self-healing nature of Leishmania major (Lm) infection, we examined the interaction between myeloid derived human DC and Lm metacyclic promastigotes (infectious-stage parasites) to model the early "natural" events of infection. We found that DC can take up Lm and, after this internalization, undergo changes in surface phenotype suggesting "maturation". Despite the intracellular location of the parasite and resultant up regulation of costimulatory and class II molecules, there was no detectable cytokine release by these Lm-harboring DC. However, using intracellular staining and flow cytometry to analyze cytokine production at the single-cell level, we found that Lm-harboring DC, but not monocytes, produce large amounts of IL-12p70 in a CD40 ligand (CD40L)-dependent manner. Finally, DC generated from mononuclear cells from patients with cutaneous leishmaniasis (Lm), once loaded with live metacyclic promastigotes, were found to reactivate autologous primed T lymphocytes and induce a CD40L-dependent IFN-gamma response. Our results link the required CD40/CD40L interactions for healing with DC-derived IL-12p70 production and provide a mechanism to explain the genesis of a protective T cell-mediated response in the face of local immune evasion within the macrophage at the site of Leishmania delivery. PMID- 10820266 TI - Localization of lung surfactant protein D on mucosal surfaces in human tissues. AB - Lung surfactant protein-D (SP-D), a collectin mainly produced by alveolar type II cells, initiates the effector mechanisms of innate immunity on binding to microbial carbohydrates. A panel of mRNAs from human tissues was screened for SP D mRNA by RT-PCR. The lung was the main site of synthesis, but transcripts were readily amplified from trachea, brain, testis, salivary gland, heart, prostate gland, kidney, and pancreas. Minor sites of synthesis were uterus, small intestine, placenta, mammary gland, and stomach. The sequence of SP-D derived from parotid gland mRNA was identical with that of pulmonary SP-D. mAbs were raised against SP-D, and one was used to locate SP-D in cells and tissues by immunohistochemistry. SP-D immunoreactivity was found in alveolar type II cells, Clara cells, on and within alveolar macrophages, in epithelial cells of large and small ducts of the parotid gland, sweat glands, and lachrymal glands, in epithelial cells of the gall bladder and intrahepatic bile ducts, and in exocrine pancreatic ducts. SP-D was also present in epithelial cells of the skin, esophagus, small intestine, and urinary tract, as well as in the collecting ducts of the kidney. SP-D is generally present on mucosal surfaces and not restricted to a subset of cells in the lung. The localization and functions of SP-D indicate that this collectin is the counterpart in the innate immune system of IgA in the adaptive immune system. PMID- 10820267 TI - Beta 2 integrins are involved in cytokine responses to whole Gram-positive bacteria. AB - Proinflammatory cytokines have an important pathophysiologic role in septic shock. CD14 is involved in cytokine responses to a number of purified bacterial products, including LPS. However, little is known of monocyte receptors involved in cytokine responses to whole bacteria. To identify these receptors, human monocytes were pretreated with different mAbs and TNF-alpha was measured in culture supernatants after stimulation with whole heat-killed bacteria. Human serum and anti-CD14 Abs significantly increased and decreased, respectively, TNF alpha responses to the Gram-negative Escherichia coli. However, neither treatment influenced responses to any of the Gram-positive bacteria tested, including group A and B streptococci, Listeria monocytogenes, and Staphylococcus aureus. Complement receptor type III (CR3 or CD18/CD11b) Abs prevented TNF-alpha release induced by heat-killed group A or B streptococci. In contrast, the same Abs had no effects when monocytes were stimulated with L. monocytogenes or S. aureus. Using either of the latter bacteria, significant inhibition of TNF-alpha release was produced by Abs to CD11c, one of the subunits of CR4. To confirm these blocking Ab data, IL-6 release was measured in CR3-, CR4-, or CD14-transfected Chinese hamster ovary cells after bacterial stimulation. Accordingly, streptococci triggered moderate IL-6 production (p < 0.05) in CR3 but not CD14 or CR4 transfectants. In contrast, L. monocytogenes and S. aureus induced IL-6 release in CR4 but not CR3 or CD14 transfectants. Collectively our data indicate that beta 2 integrins, such as CR3 and CR4, may be involved in cytokine responses to Gram-positive bacteria. Moreover, CD14 may play a more important role in responses to whole Gram-negative bacteria relative to Gram-positive ones. PMID- 10820268 TI - The role of CD40-CD154 interaction in antiviral T cell-independent IgG responses. AB - Polyomavirus (PyV) infection elicits protective T cell-independent (TI) IgG responses in T cell-deficient mice. The question addressed in this report is whether CD40 signaling plays a role in this TI antiviral IgG response. Because CD40 ligand (CD40L) can be expressed on numerous cell types in addition to activated T cells, it is possible that cells other than T cells provide CD40L to signal through CD40 on B cells and hence positively influence the antiviral TI IgG responses. In this study we show, by blocking CD40-CD40L interactions in vivo with anti-CD40L Ab treatment in TCR betaxdelta-/- mice and by using SCID mice reconstituted with CD40-/- B cells, that the lack of CD40 signaling in B cells results in a 50% decrease in TI IgG secreted in response to PyV. SCID mice reconstituted with CD40L-/- B cells also responded to PyV infection with diminished IgG secretion compared with that of SCID mice reconstituted with wild type B cells. This finding suggests that B cells may provide the CD40L for CD40 signaling in the absence of T cell help during acute virus infection. Our studies demonstrate that, although about half of the TI IgG responses to PyV are independent of CD40-CD40L interactions, these interactions occur in T cell deficient mice and enhance antiviral TI Ab responses. PMID- 10820269 TI - Secreted and membrane-associated matrix metalloproteinases of IL-2-activated NK cells and their inhibitors. AB - We have previously documented that rat IL-2-activated NK (A-NK) cells produce matrix metalloproteinase-2 (MMP-2) and MMP-9. In this study, we describe mouse A NK cell-derived MMPs, including MT-MMPs, and also TIMPs. RT-PCR analysis from cDNA of mouse A-NK cells revealed mRNA for MMP-2, MMP-9, MMP-11, MMP-13, MT1-MMP, MT2-MMP, TIMP-1, and TIMP-2. MMP-2 and MMP-9 expression was confirmed by gelatin zymography. Moreover, we report for the first time that MT-MMPs are expressed by NK cells, i.e., large granular lymphocytes as determined by both RT-PCR and Western blots. TIMP-1 expression was detected as a 29-kDa protein in Western blots. It is intriguing that TIMP-2 protein from A-NK cells was also detected as a 29-kDa protein, which is clearly different from the previously reported molecular mass of 21 kDa in mouse and human cells. In addition, inhibition of MMPs by BB-94, a selective inhibitor of MMP, significantly inhibited the ability of mouse A-NK cells to migrate through Matrigel, a model basement membrane. Taken together, these findings suggest that A-NK cells may therefore use multiple MMPs in various cellular functions, including degradation of various extracellular matrix molecules as they extravasate from blood vessels and accumulate within cancer metastases following their adoptive transfer. PMID- 10820270 TI - Genetically resistant mice lacking IL-18 gene develop Th1 response and control cutaneous Leishmania major infection. AB - IL-18 has been shown to play a critical role in the development of a Th1 response and immunity against intracellular pathogens. To determine the role of IL-18 in the development of protective immunity against Leishmania major, we have analyzed the course of cutaneous L. major in IL-18-deficient C57BL/6 mice (IL-18-/-) compared with similarly infected wild-type mice (IL-18+/+). After L. major infection, IL-18-/- mice may develop larger lesions during early phase of infection but eventually will resolve them as efficiently as IL-18+/+ mice. By 2 wk after infection, although Ag-stimulated lymph node cells from L. major infected IL-18+/+ and IL-18-/- mice produced similar levels of IFN-gamma, those from IL-18-/- mice produced significantly more IL-12 and IL-4. By 10 wk after infection, both IL-18+/+ and IL-18-/- mice had resolved L. major infection. At this time, lymph node cells from both IL-18+/+ and IL-18-/- mice produced IL-12 and IFN-gamma but no IL-4. Furthermore, administration of anti-IFN-gamma Abs to IL-18-/- mice rendered them susceptible to L. major. These results indicate that despite the role IL-18 may play in early control of cutaneous L. major lesion growth, this cytokine is not critical for development of protective Th1 response and resolution of L. major infection. PMID- 10820271 TI - Salmonella typhimurium infection and lipopolysaccharide stimulation induce similar changes in macrophage gene expression. AB - Changes in macrophage phenotype induced during infection result from the recognition of bacterial products as well as the action of bacterial virulence factors. We used the unprecedented opportunity provided by gene arrays to simultaneously study the expression of hundreds of genes during Salmonella typhimurium infection of macrophages and to assess the contribution of the bacterial virulence factor, LPS, in initiating the host responses to Salmonella. We found that S. typhimurium infection caused significant changes in the expression of numerous genes encoding chemokines, cell surface receptors, signaling molecules, and transcriptional activators at 4 h postinfection of the RAW 264.7 murine macrophage cell line. Our results revealed changes in the expression of several genes that had not been previously implicated in the host responses to S. typhimurium infection, as well as changes in the expression of several genes previously shown to be regulated by S. typhimurium infection. An overlapping spectrum of genes was expressed in response to virulent S. typhimurium and purified S. typhimurium LPS, reinforcing the major role of this surface molecule in stimulating the early response of macrophages to bacterial infection. The macrophage gene expression profile was further altered by activation with IFN-gamma, indicating that host cell responses depend on the activation state of the cell. PMID- 10820272 TI - Improving protective immunity induced by DNA-based immunization: priming with antigen and GM-CSF-encoding plasmid DNA and boosting with antigen-expressing recombinant poxvirus. AB - Intramuscular immunization with a naked DNA plasmid expressing the Plasmodium yoelii circumsporozoite protein (pPyCSP) protects mice against challenge with P. yoelii sporozoites. This protection can be improved either by coadministration of a plasmid expressing murine GM-CSF (pGMCSF) or by boosting with recombinant poxvirus expressing the PyCSP. We now report that combining these two strategies, by first mixing the priming dose of pPyCSP with pGMCSF and then boosting with recombinant virus, can substantially increase vaccine effectiveness. Not only were immune responses and protection improved but the pPyCSP dose could be lowered from 100 microg to 1 microg with little loss of immunogenicity after boost with recombinant poxvirus. Comparing mice primed by the 1-microg doses of pPyCSP plus 1 microg pGMCSF with mice primed by 1-microg doses of pPyCSP alone, the former were better protected (60% vs 0) and had higher concentrations of Abs (titers of 163, 840 vs 5, 120 by indirect fluorescent Ab test against sporozoites), more ex vivo CTL activity (25% vs 7% specific lysis), and more IFN gamma-secreting cells by enzyme-linked immunospot assay (1460 vs 280 IFN-gamma spot-forming cells/106 cells). Priming with plasmid vaccine plus pGMCSF and boosting with recombinant poxviruses strongly improves the immunogenicity and protective efficacy of DNA vaccination and allows for significant reduction of dose. PMID- 10820273 TI - CD40 ligand (CD154) enhances the Th1 and antibody responses to respiratory syncytial virus in the BALB/c mouse. AB - CD40 ligand (CD40L) is a cell surface costimulatory molecule expressed mainly by activated T cells. CD40L is critically important for T-B cell and T cell dendritic cell interactions. CD40L expression promotes Th1 cytokine responses to protein Ags and is responsible for Ig isotype switching in B cells. Respiratory syncytial virus (RSV) is an important pathogen of young children and the elderly, which causes bronchiolitis and pneumonia. Studies of mice infected with RSV suggest that a Th2 cytokine response may be responsible for enhanced pulmonary disease. To investigate the effect CD40L has on RSV immunity, mice were infected simultaneously with RSV and either an empty control adenovirus vector or one expressing CD40L or were coimmunized with plasmid DNA vectors expressing CD40L and RSV F and/or G proteins and subsequently challenged with RSV. The kinetics of the intracellular and secreted cytokine responses, the cytotoxic T lymphocyte precursor frequency, NO levels in lung lavage, rates of virus clearance, and anti RSV Ab titers were determined. These studies show that coincident expression of CD40L enhances the Th1 (IL-2 and IFN-gamma) cytokine responses, increases the expression of TNF-alpha and NO, accelerates virus clearance, and increases the anti-F and anti-G Ab responses. These data suggest that CD40L may have the adjuvant properties needed to optimize the safety and efficacy of RSV vaccines. PMID- 10820274 TI - Vascular endothelial growth factor expression and regulation of murine collagen induced arthritis. AB - We have examined the expression and function of the angiogenic factor, vascular endothelial growth factor (VEGF) during the evolution of type II collagen-induced arthritis (CIA). Biologically active VEGF was expressed along a time course that paralleled the expression of two specific VEGF receptors, Flk-1 and Flt-1, and the progression of joint disease. Moreover, levels of VEGF expression correlated with the degree of neovascularization, as defined by vWF levels, and arthritis severity. Macrophage- and fibroblast-like cells, which infiltrated inflamed sites and were then activated by other inflammatory mediators, are probably important sources of VEGF and may thus regulate angiogenesis during the development of CIA. Administration of anti-VEGF antiserum to CIA mice before the onset of arthritis delayed the onset, reduced the severity, and diminished the vWF content of arthritic joints. By contrast, administration of anti-VEGF antiserum after the onset of the disease had no effect on the progression or ultimate severity of the arthritis. These data suggest that VEGF plays a crucial role during an early stage of arthritis development, affecting both neovascularization and the progression of experimentally induced synovitis. PMID- 10820275 TI - Human blood monocytes interact with type I collagen through alpha x beta 2 integrin (CD11c-CD18, gp150-95). AB - Human blood monocytes are attracted into connective tissues during early steps of inflammation and wound healing, and locally interact with resident cells and extracellular matrix proteins. We studied the effects of type I collagen on monocyte adhesion and superoxide anion production, using human monocytes elutriated from peripheral blood and type I collagen obtained from rat tail tendon. Both acid-soluble and pepsin-digested type I collagens promoted the adhesion of monocytes, whereas only acid-soluble collagen with intact telopeptides induced the production of superoxide. Adhesion and activation of monocytes on acid-soluble type I collagen depended on the presence of divalent cations. mAbs directed against integrin subunits CD11c and CD18 specifically inhibited adhesion and activation of monocytes on type I collagen. Protein membrane extracts obtained from monocytes were submitted to affinity chromatography on collagen I-Sepharose 4B, and analyzed by Western blotting using specific anti-integrin subunit Abs. In the case of both acid-soluble and pepsin digested collagens, two bands were revealed with mAbs against CD11c and CD18 integrin subunits. Our results demonstrate that monocytes interact with type I collagen through CD11c-CD18 (alpha x beta 2) integrins, which promote their adhesion and activation. For monocyte activation, specific domains of the type I collagen telopeptides are necessary. Interactions between monocytes and collagen are most likely involved in the cascade of events that characterize the initial phases of inflammation. PMID- 10820276 TI - Expression of CXCR4 in eosinophils: functional analyses and cytokine-mediated regulation. AB - We examined the expression of transcripts of a panel of chemokine receptors in human eosinophils and found intense constitutive expression of CXCR4 mRNA. Although surface CXCR4 protein was hardly detectable in the peripheral blood or freshly isolated eosinophils, surface expression of CXCR4 became gradually apparent during incubation at 37 degrees C. In contrast, the level of CCR3 expression was virtually unchanged during the incubation. Stromal cell-derived factor-1alpha (SDF-1alpha), the natural ligand of CXCR4, elicited an apparent Ca2+ influx in these cells and induced a strong migratory response comparable to that by eotaxin. The surface expression of CXCR4 in eosinophils was up-regulated by IFN-gamma, TNF-alpha, and TGF-beta while it was down-regulated by IL-4 and eosinophil-directed hemopoietins such as IL-5. The CXCR4 expression did not always parallel the apoptotic changes in cytokine-treated eosinophils. In contrast to IL-4 and IFN-gamma, IL-5 potently reduced the level of CXCR4 mRNA. It seems unlikely that CXCR4 is fundamentally involved in the pathogenesis of allergic disorders by inducing the migration of eosinophils toward inflammatory sites, because a Th2-dominant state down-regulates eosinophil CXCR4 expression. However, CXCR4 may affect the size of the mobilizable pool by holding eosinophils at noninflamed tissues. Th2-dominant state may favor the liberation of eosinophils by down-regulating CXCR4 expression. The interplay between CXCR4 and SDF-1alpha in eosinophils potentially plays an important role in the accumulation of these cells at the allergic inflammatory sites. PMID- 10820277 TI - Requirement of a novel upstream response element in respiratory syncytial virus induced IL-8 gene expression. AB - Respiratory syncytial virus (RSV) produces intense pulmonary inflammation, in part, through its ability to induce chemokine synthesis in infected airway epithelial cells. In this study, we compare mechanisms for induction of the CXC chemokine IL-8, in human type II alveolar (A549) cells by RSV infection and by stimulation with the cytokine TNF. Promoter deletion and mutagenesis experiments indicate that although the region from -99 to -54 nt is sufficient for TNF induced IL-8 transcription, this region alone is not sufficient for RSV-induced IL-8 transcription. Instead, RSV requires participation of a previously unrecognized element, spanning from -162 to -132 nt, that we term the RSV response element (RSVRE), and a previously characterized element at -132 to -99 nt, containing an AP-1 binding site. RSV infection of A549 cells induces increased RSVRE- and AP-1-binding activities and increased synthesis of IFN regulatory factor-1 protein, which is present in the RSVRE-binding complex. These data confirm that the IL-8 gene enhancers are controlled in a stimulus-specific fashion and participation of distinct promoter elements is required to activate gene transcription. These observations are important for rational design of inhibitors of RSV-induced lung inflammation. PMID- 10820278 TI - Th2 cell membrane factors in association with IL-4 enhance matrix metalloproteinase-1 (MMP-1) while decreasing MMP-9 production by granulocyte macrophage colony-stimulating factor-differentiated human monocytes. AB - Monocytes/macrophages are directly involved in tissue remodeling and tissue destruction through the release of matrix metalloproteinases (MMP). In the present study, we examined the effect mediated by contact of polarized Th cells with mononuclear phagocytes on the production of MMP-1, MMP-9, and their inhibitor. Plasma cell membranes from Ag-activated Th1 and Th2 cells were potent inducers of MMP-1 production by THP-1 cells. Cell membrane-associated TNF was found to be only partially involved in MMP-1 induction by both Th1 and Th2 cells. In Th2 cells exclusively, membrane-associated IL-4 induced MMP-1 production by THP-1 cells. This membrane-associated IL-4 effect was additive to that of TNF and was specifically observed on MMP-1 as MMP-9 production was concomitantly inhibited. Similarly, soluble IL-4 induced THP-1 cells to produce MMP-1, its effect proving additive to that of soluble TNF and to that of cell membranes of mitogen-activated HUT-78 cells. Its activity was blocked by IL-4 neutralization, and was unaffected by the presence of indomethacin. These effects on THP-1 cells were observed at protein and mRNA levels. Although inhibitory on freshly isolated peripheral blood monocytes, soluble IL-4 enhanced T cell-induced MMP-1 and inhibited MMP-9 production both at protein and mRNA levels in monocytes cultured for 7 days in the presence of GM-CSF. Thus, in contrast with previously reported effects, Th2 and IL-4 specifically induce MMP-1 production by mononuclear phagocytes at various stages of differentiation. This IL-4 activity may be relevant to pathological conditions dominated by Th2 inflammatory responses, resulting in tissue remodeling and destruction. PMID- 10820279 TI - Differential ability of exogenous chemotactic agents to disrupt transendothelial migration of flowing neutrophils. AB - Neutrophils migrate through endothelium using an ordered sequence of adhesive interactions and activating signals. To investigate the consequences of disruption of this sequence, we characterized adhesion and migration of neutrophils perfused over HUVEC that had been treated with TNF-alpha for 4 h and evaluated changes caused by exogenously added chemotactic agents. When HUVEC were treated with 2 U/ml TNF, flowing neutrophils adhered, with the majority rolling and relatively few migrating through the monolayer. If fMLP, IL-8, zymosan activated plasma (a source of activated complement factor C5a), epithelial cell derived neutrophil-activating peptide (ENA-78), or growth-regulating oncogene, GRO-alpha, was perfused over these neutrophils, they stopped rolling and rapidly migrated over the monolayer, but did not penetrate it. When HUVEC were treated with 100 U/ml TNF, the majority of adherent neutrophils transmigrated. If neutrophils were treated with fMLP, IL-8, C5a, ENA-78, or GRO-alpha just before perfusion over this HUVEC, transmigration, but not adhesion, was abolished. However, when platelet-activating factor was used to activate neutrophils, migration through HUVEC treated with 100 U/ml TNF was not impaired, and migration through HUVEC treated with 2 U/ml TNF was actually increased. Transmigration required ligation of CXC chemokine receptor-2 on neutrophils, and differential desensitization of this receptor (e.g., by fMLP but not platelet-activating factor) may explain the pattern of disruption of migration. Thus, transmigration may require presentation of the correct activators in the correct sequence, and inappropriate activation (e.g., by systemic activators) could cause pathological accumulation of neutrophils in the vessel lumen. PMID- 10820280 TI - Molecular mechanisms of increased nitric oxide (NO) in asthma: evidence for transcriptional and post-translational regulation of NO synthesis. AB - Evidence supporting increased nitric oxide (NO) in asthma is substantial, although the cellular and molecular mechanisms leading to increased NO are not known. Here, we provide a clear picture of the events regulating NO synthesis in the human asthmatic airway in vivo. We show that human airway epithelium has abundant expression of NO synthase II (NOSII) due to continuous transcriptional activation of the gene in vivo. Individuals with asthma have higher than normal NO concentrations and increased NOSII mRNA and protein due to transcriptional regulation through activation of Stat1. NOSII mRNA expression decreases in asthmatics receiving inhaled corticosteroid, treatment effective in reducing inflammation in asthmatic airways. In addition to transcriptional mechanisms, post-translational events contribute to increased NO synthesis. Specifically, high output production of NO is fueled by a previously unsuspected increase in the NOS substrate, l -arginine, in airway epithelial cells of asthmatic individuals. Finally, nitration of proteins in airway epithelium provide evidence of functional consequences of increased NO. In conclusion, these studies define multiple mechanisms that function coordinately to support high level NO synthesis in the asthmatic airway. These findings represent a crucial cornerstone for future therapeutic strategies aimed at regulating NO synthesis in asthma. PMID- 10820281 TI - Thiol-reactive metal compounds inhibit NF-kappa B activation by blocking I kappa B kinase. AB - Gold compounds are used in the treatment of rheumatoid arthritis. NF-kappa B is a transcription factor implicated in the expression of many inflammatory genes. NF kappa B is activated by signal-induced phosphorylation and subsequent degradation of inhibitory I kappa B (inhibitory protein that dissociates from NF-kappa B) proteins, and a multisubunit I kappa B kinase (IKK) has been identified previously. We tested the effect of various gold compounds on the activation of NF-kappa B and IKK in LPS-stimulated RAW 264.7 mouse macrophages. A lipophilic gold compound, auranofin, suppressed the LPS-induced increase of nuclear kappa B binding activity, degradation of I kappa B proteins, and IKK activation. Auranofin also blocked IKK activation induced by TNF and PMA/ionomycin, suggesting that the target of auranofin action is common among these diverse signal pathways. In vitro IKK activity was suppressed by addition of hydrophilic gold compounds, such as aurothiomalate, aurothioglucose, and AuCl3. Other thiol reactive metal ions such as zinc and copper also inhibited IKK activity in vitro, and induction of IKK in LPS-stimulated macrophages. In vitro IKK activity required the presence of reducing agent and was blocked by addition of thiol group-reactive agents. Two catalytic subunits of IKK complex, IKK alpha and IKK beta, were both inhibited by these thiol-modifying agents, suggesting the presence of a cysteine sulfhydryl group in these subunits, which is critical for enzyme activity. The antiinflammatory activity of gold compounds in the treatment of rheumatoid arthritis may depend on modification of this thiol group by gold. PMID- 10820282 TI - NF-kappa B regulates VCAM-1 expression on fibroblast-like synoviocytes. AB - Expression of VCAM-1 on synovial fibroblasts is a clinical hallmark of rheumatoid arthritis. The interaction of VCAM-1 and its integrin receptor very late Ag-4 is believed to be critically involved in the recruitment and retention of immune cells in the inflamed joints. To study the regulation of VCAM-1 in synovial fibroblasts, fibroblast-like synoviocytes (FLS) were isolated from the knee joints of normal mice and passaged repeatedly to obtain a homogeneous cell population. We have found that VCAM-1 is constitutively expressed on mouse FLS (mFLS) and that its surface expression is further increased after exposure to TNF alpha. Nuclear translocation of transcription factor NF-kappa B including P50/P50 homodimer and P65/P50 heterodimer was activated by TNF-alpha treatment. In mFLS stably expressing a dominant-negative mutant of the inhibitory protein I-kappa B alpha- (mI-kappa B), which does not undergo proteolytic degradation, NF-kappa B remains in the cytosol and its activation in response to TNF-alpha is abolished. VCAM-1 protein expression after TNF-alpha stimulation was blocked in cells expressing the mI-kappa B. This effect is likely due to the loss of NF-kappa B mediated transcription of VCAM-1, because the 5-fold increase in mRNA levels in response to TNF-alpha is absent in the mutant cells. To confirm these findings, we transduced mFLS with an adenoviral vector containing the mI-kappa B transgene. VCAM-1 expression was also blocked by mI-kappa B in this system, whereas cells transduced with a control adenoviral vector remained responsive to TNF-alpha. These results indicate that NF-kappa B mediates TNF-alpha-induced VCAM-1 expression on mFLS. PMID- 10820283 TI - Differential expression and regulation of toll-like receptors (TLR) in human leukocytes: selective expression of TLR3 in dendritic cells. AB - Members of the Toll-like receptor (TLR) family probably play a fundamental role in pathogen recognition and activation of innate immunity. The present study used a systematic approach to analyze how different human leukocyte populations express specific transcripts for the first five characterized TLR family members. TLR1 was expressed in all leukocytes examined, including monocytes, polymorphonuclear leukocytes, T and B cells, and NK cells. In contrast TLR2, TLR4, and TLR5 were expressed in myelomonocytic elements. Exposure to bacterial products, such as LPS or lipoarabinomannan, or to proinflammatory cytokines increased TLR4 expression in monocytes and polymorphonuclear leukocytes, whereas IL-10 blocked this effect. TLR3 was only expressed in human dendritic cells (DC) wherein maturation induced by bacterial products or cytokines was associated with reduced expression. TLR3 mRNA expression was detected by in situ hybridization in DC and lymph nodes. These results demonstrate that TLR1 through TLR5 mRNAs are differentially expressed and regulated in human leukocytes. In particular, expression of TLR3 transcripts is restricted to DC that are the only elements which express the full TLR repertoire. These data suggest that TLR can be classified based on expression pattern as ubiquitous (TLR1), restricted (TLR2, TLR4, and TLR5 in myelomonocytic cells), and specific (TLR3 in DC) molecules. PMID- 10820284 TI - Prevention of experimental colitis in SCID mice reconstituted with CD45RBhigh CD4+ T cells by blocking the CD40-CD154 interactions. AB - Increased expression of CD40 and CD40 ligand (CD40L or CD154) has been found in inflamed mucosa of human inflammatory bowel disease (IBD), and interactions between these molecules seem to be involved in local cytokine production by macrophages. However, the precise role of CD40 signaling in the pathogenesis of IBD is still poorly understood. The aim of the present study was to investigate the in vivo relevance of CD40 signaling in experimental colitis in SCID mice reconstituted with syngeneic CD45RBhighCD4+ T cells. The results demonstrated that CD40+ and CD40L+ cells as well as their mRNA levels were significantly increased in inflamed mucosa. Administration of anti-CD40L neutralizing mAb over an 8-wk period starting immediately after CD45RBhighCD4+ T cell reconstitution completely prevented symptoms of wasting disease. Intestinal mucosal inflammation was effectively prevented, as revealed by abrogated leukocyte infiltration and decreased CD54 expression and strongly diminished mRNA levels of the proinflammatory cytokines IFN-gamma, TNF, and IL-12. When colitic SCID mice were treated with anti-CD40L starting at 5 wk after T cell transfer up to 8 wk, this delayed treatment still led to significant clinical and histological improvement and down-regulated proinflammatory cytokine secretion. These data suggest that the CD40-CD40L interactions are essential for the Th1 inflammatory responses in the bowel in this experimental model of colitis. Blockade of CD40 signaling may be beneficial to human IBD. PMID- 10820285 TI - Simian immunodeficiency virus (SIV)-specific CTL are present in large numbers in livers of SIV-infected rhesus monkeys. AB - The immunopathogenesis of AIDS-associated hepatitis was explored in the SIV/rhesus monkey model. The livers of SIV-infected monkeys showed a mild hepatitis, with a predominantly CD8+ T lymphocyte infiltration in the periportal fields and sinusoids. These liver-associated CD8+ T cells were comprised of a high percentage of SIV-specific CTL as defined by MHC class I/Gag peptide tetramer binding and Gag peptide epitope-specific lytic activity. There was insufficient viral replication in these livers to account for attracting this large number of functional virus-specific CTL to the liver. There was also no evidence that the predominant population of CTL were functionally end-stage cells trapped in the liver and destined to undergo apoptotic cell death in that organ. Interestingly, we noted that liver tetramer-binding cells showed an increased expression of CD62L, an adhesion molecule usually only rarely expressed on tetramer-binding cells. This observation suggests that the expression of specific adhesion molecules by CTL might facilitate the capture of these cells in the liver. These results demonstrate that functional SIV-specific CD8+ T cells are present in large numbers in the liver of chronically SIV-infected monkeys. Thus, the liver may be a trap for virus-specific cytotoxic T cells. PMID- 10820286 TI - Novel protein transfection of primary rat cortical neurons using an antibody that penetrates living cells. AB - An Ab-based system to deliver functional proteins into neurons was developed using the murine mAb, mAb 3E10. This was achieved by covalently conjugating catalase to the Ab so that the conjugate retained high activity for the degradation of hydrogen peroxide. Three-dimensional fluorescence microscopy was used to demonstrate penetration of the Ab into the nucleus of living primary cortical neurons. The Ab conjugate localized in both the cytoplasm and nucleus. Retention of catalase activity after penetration and distribution of conjugate was demonstrated by reduction in cell death following exposure of treated neurons to hydrogen peroxide. These studies illustrate the potential of this method for the intracellular delivery of therapeutic proteins. PMID- 10820287 TI - Intraallograft chemokine RNA and protein during rejection of MHC-matched/multiple minor histocompatibility-disparate skin grafts. AB - Chemokines direct leukocyte recruitment into sites of tissue inflammation and may facilitate recruitment of leukocytes into allografts following transplantation. Although the expression of chemokines during rejection of MHC-disparate allografts has been examined, chemokine expression in MHC-matched/multiple minor histocompatibility Ag-disparate allografts has not been tested. The intraallograft RNA expression of several C-X-C and C-C chemokines was tested during rejection of full thickness skin grafts from B10. D2 donors on control Ig , anti-CD4 mAb-, and anti-CD8 mAb-treated BALB/c recipients. In all recipients, two patterns of intragraft chemokine expression were observed during rejection of these grafts: 1) macrophage-inflammatory protein-1alpha, macrophage-inflammatory protein-1beta, GRO-alpha (KC), JE, and IFN-gamma-inducible protein (IP-10) were expressed at equivalent levels in allo- and isografts for 2-4 days posttransplant and then returned to low or undetectable levels; and 2) IP-10 and monokine induced by IFN-gamma (Mig) were expressed in the allografts 3 days before rejection was completed, suggesting a possible role in recruiting primed T cells into the allograft. Three days before completion of rejection, intraallograft IP 10 protein was restricted to the epidermis, whereas Mig was located in the lower dermis and associated with the intense infiltration of mononuclear cells. Treatment of B10.D2 recipients with rabbit antiserum to Mig, but not to IP-10, delayed rejection of the allografts 3-4 days. The results suggest that Mig mediates optimal recruitment of T cells into MHC-matched/multiple minor histocompatibility Ag-disparate allografts during rejection. PMID- 10820288 TI - Threshold signaling of human Th0 cells in activation and anergy: modulation of effector function by altered TCR ligand. AB - Molecular interactions between TCR and its natural ligand, in the presence of costimulatory signals, elicit T cell effector functions, whereas subtle changes in the structure of antigenic peptides may induce only selected T cell effector function including anergy. In this study, we have investigated the immunological activity of an altered TCR ligand (p 2, 28-40A34,36) derived from the immunodominant T cell epitope of the group 2 allergen of house dust mite, in which residues at positions 34 and 36 were substituted by alanine. Elevated IFN gamma synthesis was induced by equimolar concentrations of the analogue compared with native peptide (p 2, 28-40) and was paralleled by increased down-regulation of cell surface CD3. IL-5 and IL-10 production exhibit the same sensitivity to both peptides, implying that the induction of T cell effector functions are not all proportional to TCR occupancy. Both native peptide and the analogue bound to MHC class II (DRB1*1101) molecules with similar affinities. Furthermore, p 2, 28 40A34,36 induced T cell anergy at lower concentrations than native peptide. During the induction of anergy, TGF-beta production was comparable for both peptides, whereas IL-10 secretion was markedly increased but more so in response to p 2, 28-40A34,36. Membrane expression of costimulatory ligands CD80 and CD86 was similar for native peptide and p 2, 28-40A34,36 and increased in activation, whereas only CD86 was elevated during anergy. The modulation of T cell effector function with altered TCR ligands may have practical applications in reprogramming allergic inflammatory responses through the induction of T cell anergy and/or the promotion of Th1 cytokines. PMID- 10820289 TI - Cytolytic T lymphocytes raised against a human bladder carcinoma recognize an antigen encoded by gene MAGE-A12. AB - Human bladder carcinoma line LB831-BLC expresses several distinct Ags that are recognized by different autologous CTL. Here, we show that one of these Ags is presented by HLA-Cw7 and encoded by gene MAGE-A12. This is the first time that CTL directed against a MAGE-encoded Ag have been derived from the lymphocytes of a patient with cancer other than melanoma. This new Ag was found to be nonapeptide VRIGHLYIL, corresponding to position 170-178 of the MAGE-A12 protein. Gene MAGE-A12 is silent in normal tissues except in male germline cells, which do not express HLA molecules. It is expressed in 26-62% of melanomas, infiltrating bladder carcinomas, lung carcinomas, esophageal carcinomas, and head and neck carcinomas. Because HLA-Cw7 is present in 43% of Caucasians, this new Ag is shared by many tumors and should be a useful target for cancer immunotherapy. PMID- 10820290 TI - Costimulation by B7-1 and B7-2 is required for autoimmune disease in MRL-Faslpr mice. AB - Autoimmune lupus nephritis is dependent on infiltrating autoreactive leukocytes and Igs. B7 costimulatory molecules (B7-1 and B7-2) provide signals essential for T cell activation and Ig class switching. In MRL-Faslpr mice, a model of human lupus, although multiple tissues are targeted for autoimmune injury, nephritis is fatal. We identified intrarenal B7-1 and B7-2 expression, restricted to kidney infiltrating leukocytes, before and increasing with progressive nephritis in MRL Faslpr mice. Thus, we hypothesized that the B7 pathway is required for autoimmune disease in MRL-Faslpr mice. To investigate the role of B7 costimulatory molecules in this autoimmune disease, we generated a MRL-Faslpr strain deficient in B7-1 and B7-2. Strikingly, MRL-Faslpr mice lacking both B7 costimulators do not develop kidney (glomerular, tubular, interstitial, vascular) pathology, or proteinuria, and survive far longer. Intrarenal downstream effector transcripts (IFN-gamma, IL-12, monocyte chemoattractant protein-1, CSF-1) linked to nephritis remained at normal levels compared with wild-type mice. Skin lesions and lymphoid enlargement characteristic of MRL-Faslpr mice were diminished in B7-1/B7-2 deficient MRL-Faslpr mice. B7-1/B7-2-deficient MRL-Faslpr mice did not develop leukocytic infiltrates, elevated serum IgG and isotypes (G1,G2b,G3), autoantibodies, and intrarenal IgG deposits. Our findings demonstrate that B7-1 and B7-2 costimulatory pathways are critical to the pathogenesis of autoimmune lupus. PMID- 10820291 TI - High frequency of autologous anti-melanoma CTL directed against an antigen generated by a point mutation in a new helicase gene. AB - We have identified an Ag recognized by autologous CTL on the melanoma cells of a patient who enjoyed an unusually favorable clinical evolution. The antigenic peptide, which is presented by HLA-A28 molecules, is encoded by a mutated sequence in a new gene. This gene, which was named MUM-3, is expressed ubiquitously and shows homology with the RNA helicase gene family. Limiting dilution analysis indicated that at least 0.15% of the blood CD8 T cells were tumor-specific CTL precursors. The MUM-3 Ag was recognized by 90% of these CTL, indicating that it is the dominant target Ag of the tumor-specific CTL response. The high frequency of anti-MUM-3 CTL was confirmed with tetramers of soluble HLA A28 molecules loaded with the antigenic peptide. MUM-3 tetramers stained 1.2% of blood CD8 cells, a frequency that has never been reported for T cells directed against a strictly tumor-specific Ag. To confirm these results, the CD8 T cells that were clearly labeled with tetramers were restimulated in clonal conditions. About 90% of these cells proliferated, and all the resulting clones proved lytic and MUM-3 specific. By improving the conditions used for the in vitro restimulation of CTL precursors by the tumor cells, the same frequency could be obtained in limiting dilution analysis. These results show that some cancer patients have a high frequency of circulating CTL that are directed against a strictly tumor-specific Ag. These CTL are responsive to restimulation in vitro and are easily detected with tetramers. Such responses may therefore be an achievable goal for therapeutic vaccination with tumor-specific Ags. PMID- 10820292 TI - IL-18 prevents the development of chronic graft-versus-host disease in mice. AB - The development of chronic graft-versus-host disease (GVHD), which is induced by the transfer of DBA/2 spleen cells into (C57BL/6 x DBA/2)F1 (BDF1) mice, is closely related to diminished donor anti-host CTL activity and host B cell hyperactivation. Therefore, an approach which activates donor CD8+ T cells or suppresses donor CD4+ T cell-host B cell interaction may have clinical utility in the treatment of chronic GVHD. We have previously demonstrated that IL-18 induces the development of naive CD8+ T cells into type I effector cells in DBA/2 anti BDF1 MLC. In this paper we examined the effect of IL-18 administration on the development of chronic GVHD in mice. The treatment was started before or after the onset of clinical evidence of the disease. Regardless of the treatment schedule, IL-18 significantly decreased immunological parameters indicative of chronic GVHD, such as elevated serum IgG antinuclear Abs, IgG1, and IgE levels, and host B cell numbers and their activation. Importantly, IL-18-treated mice did not show the same acute GVHD-like symptoms reported for IL-12 treatment, because there was no weight loss, death, or severe immunodeficiency as indicated by a decrease in IL-2 and IFN-gamma production by Con A-stimulated spleen cells. In contrast, IL-18 treatment partially but significantly restored the production of these cytokines. Data further suggested that these IL-18-mediated therapeutic effects may be due to the induction of donor CD8+ CTL, the decrease in donor CD4+ T cell numbers, and a down-regulation of host B cell MHC class II expression. Thus, our results suggest that IL-18 has beneficial effects in the prevention and treatment of chronic GVHD. PMID- 10820293 TI - Exceptional resistance of human H2 glioblastoma cells to complement-mediated killing by expression and utilization of factor H and factor H-like protein 1. AB - Of over 20 nucleated cell lines we have examined to date, human H2 glioblastoma cells have turned out to be the most resistant to complement-mediated cytolysis in vitro. H2 cells expressed strongly the membrane attack complex inhibitor protectin (CD59), moderately CD46 (membrane cofactor protein) and CD55 (decay accelerating factor), but no CD35 (complement receptor 1). When treated with a polyclonal anti-H2 Ab, anti-CD59 mAb, and normal human serum, only 5% of H2 cells became killed. Under the same conditions, 70% of endothelial-like EA.hy 926 cells and 40% of U251 control glioma cells were killed. A combined neutralization of CD46, CD55, and CD59 increased H2 lysis only minimally, demonstrating that these complement regulators are not enough to account for the resistance of H2 cells. After treatment with Abs and serum, less C5b-9 was deposited on H2 than on U251 and EA.hy 926 cell lines. A reason for the exceptional resistance of H2 cells was revealed when RT-PCR and protein biochemical methods showed that the H2 cells, unlike the other cell lines tested, actively produced the soluble complement inhibitors factor H and factor H-like protein 1. H2 cells were also capable of binding human factor H from the fluid phase to their cell surface and promoted the cleavage of C3b to its inactive form iC3b more efficiently than U251 and EA.hy 926 cells. In accordance, anti-factor H mAbs enhanced killing of H2 glioblastoma cells. Taken together, our results show that production and binding of factor H and factor H-like protein 1 is a novel mechanism that these malignant cells utilize to escape complement-mediated killing. PMID- 10820294 TI - A combined ischemic compression and spinal manipulation in the treatment of fibromyalgia: a preliminary estimate of dose and efficacy. AB - OBJECTIVES: To provide preliminary information on whether a regimen of 30 chiropractic treatments that combines ischemic compression and spinal manipulation effectively reduces the intensity of pain, sleep disturbance, and fatigue associated with fibromyalgia. In addition, to study the dose-response relation and identify the baseline characteristics that may serve as predictors of outcome. DESIGN: Subjects were assessed with self-administered questionnaires taken at baseline, after 15 and 30 treatments, and 1 month after the end of the treatment trial. SETTING: Private practice. METHODS: Participating subjects were adult members of a regional Fibromyalgia Association. Participating subjects had fibromyalgia for more than 3 months. They received 30 treatments including ischemic compression and spinal manipulation. The 3 outcomes being evaluated were pain intensity, fatigue level, and sleep quality. A minimum 50 improvement in pain intensity from baseline to the end of the treatment trial was needed to include the patient in the respondent category. RESULTS: Fifteen women (mean age 51.1 years) completed the trial. A total of 9 (60) patients were classified as respondents. A statistically significant lessening of pain intensity and corresponding improvement in quality of sleep and fatigue level were observed after 15 and 30 treatments. After 30 treatments, the respondents showed an average lessening of 77.2 (standard deviation = 12.3) in pain intensity and an improvement of 63.5 (standard deviation = 31.6) in sleep quality and 74.8 (standard deviation = 23. 1) in fatigue level. The improvement in the 3 outcome measures was maintained after 1 month without treatment. Subjects with less than 35 improvement after 15 treatments did not show a satisfactory response after 30 treatments. A trend, determined as not statistically significant, suggests that older subjects with severe and more chronic pain and a greater number of tender points respond more poorly to treatment. CONCLUSION: This study suggests a potential role for chiropractic care in the management of fibromyalgia. A randomized clinical trial should be conducted to test this hypothesis. PMID- 10820295 TI - Reliability of chiropractic methods commonly used to detect manipulable lesions in patients with chronic low-back pain. AB - OBJECTIVE: To assess the intraexaminer and interexaminer reliability of a multidimensional spinal diagnostic method commonly used by chiropractors. DESIGN: An intraexaminer and interexaminer Latin square, repeated measures reliability study. The techniques of diagnosis under investigation included visual postural analysis, pain description by the patient, plain static erect x-ray film of the lumbar spine, leg length discrepancy, neurologic tests, motion palpation, static palpation, and orthopedic tests. PARTICIPANTS: Three experienced chiropractors examined 19 patients, and 2 experienced chiropractors examined 10 and 9 patients, respectively, who were suffering from chronic mechanical low-back pain. RESULTS: Intraexaminer reliability of the decision to manipulate a certain spinal segmental level was moderate (kappa = 0.47). The interexaminer agreement pooled across all spinal joints indicated fair agreement (kappa = 0.27). Interexaminer reliability for individual examiner pairs for the L4/L5 segmental level was slight (kappa = 0.09). At the L5/S1 level, the interexaminer reliability was fair (kappa = 0.25). For the sacroiliac joints, interexaminer reliability was slight (kappa = 0.04 and 0.14). CONCLUSION: This study of commonly used chiropractic diagnostic methods in patients with chronic mechanical low-back pain to detect manipulable lesions in the lower thoracic spine, lumbar spine, and the sacroiliac joints has revealed that the measures are not reproducible. The implementation of these examination techniques alone should not be seen by practitioners to provide reliable information concerning where to direct a manipulative procedure in patients with chronic mechanical low-back pain. PMID- 10820296 TI - Patient characteristics, practice activities, and one-month outcomes for chronic, recurrent low-back pain treated by chiropractors and family medicine physicians: a practice-based feasibility study. AB - BACKGROUND: Chronic low-back pain is a significant public health problem for which few therapies are supported by predictable outcomes. In this report, practice activities and 1-month outcomes data are presented for 93 chiropractic patients and 45 medical patients with chronic, recurrent low-back pain. DESIGN: A prospective, observational, community-based feasibility study involving chiropractors and family medicine physicians. SETTING: Forty private chiropractic clinics, the outpatient clinic of the Department of Family Medicine at Oregon Health Sciences University, and 5 other Portland area family medicine clinics. OUTCOMES MEASURES: The main outcome measures were pain severity, functional disability, sensory and affective pain quality at 1 month, and patient satisfaction assessed at 7 to 10 days and at 1 month. RESULTS: Although differences were noted in age, sex, education, and employment, the patients were closely matched at baseline with respect to frequency, severity, and type of low back pain and the psychosocial dimensions of general health. The treatment of choice for chiropractors was spinal manipulation and physical therapy modalities; for medical physicians antiinflammatory agents were most frequently used. Chiropractic patients averaged 4 visits, and medical patients averaged 1 visit. On average, chiropractic patients showed improvement across all outcomes: 31% change in pain severity, 29% in functional disability, 36% in sensory pain quality, and 57% in affective pain quality. Medical patients showed minimal improvement in pain severity (6%) and functional disability (1%) and showed deterioration in the sensory (29%) and affective (26%) dimensions of pain quality. Satisfaction scores were higher for chiropractic patients. Outcomes for medical patients were heavily dependent on psychosocial status at baseline. CONCLUSION: Patients with chronic low-back pain treated by chiropractors show greater improvement and satisfaction at 1 month than patients treated by family physicians. Nonclinical factors may play an important role in patient progress. Findings from the Health Resources and Services Administration-funded project will include a report on the influence of practice activities, including more frequent visits by chiropractic patients, on the clinical course of low-back pain and patient outcomes. (J Manipulative Physiol Ther 2000;23:239-45). PMID- 10820297 TI - Use of a mental rotation reaction-time paradigm to measure the effects of upper cervical adjustments on cortical processing: a pilot study. AB - OBJECTIVES: To investigate the potential usefulness of a mental rotation paradigm in providing an objective measure of spinal manipulative therapy. To determine if cortical processing, as indicated by response time to a mental rotation reaction time task, is altered by an upper cervical toggle recoil adjustment. DESIGN: Prospective, double-blind, randomized, controlled trial. SETTING: Chiropractic college clinical training facility. PARTICIPANTS: Thirty-six chiropractic student volunteers with clinical evidence of upper cervical joint dysfunction. INTERVENTION: Participants in the experimental group received a high-velocity, low-amplitude upper cervical adjustment. A non-intervention group was used to control for improvement in the mental rotation task as a result of practice effects. OUTCOME MEASURES: Reaction time was measured for randomly varying angular orientations of an object appearing either as normal or mirror-reversed on a computer screen. RESULTS: The average decrease in mental rotation reaction time for the experimental group was 98 ms, a 14.9% improvement, whereas the average decrease in mental rotation reaction time for the control group was 58 ms, an 8. 0 improvement. The difference scores after the intervention time were significantly greater for the experimental group compared with the control group, as indicated by a one-tailed, 2-sample, equal variance Student t test, (P < 05). CONCLUSION: The results of this study have demonstrated a significant improvement in a complex reaction-time task after an upper cervical adjustment. These results provide evidence that upper cervical adjustment may affect cortical processing. PMID- 10820298 TI - Biomechanic evaluation of the Rola Stretcher as a passive distraction device. AB - INTRODUCTION: Many devices have been marketed claiming to increase the mobility of the articulations of the spine with active or passive distraction. In this study, the Rola Stretcher (Unique Relief, Inc, Davenport, Iowa) and an earlier version, the True Back II (True Back, Inc, Clearwater, Fla), were evaluated to see if they have a measurable biomechanic effect on the spine. METHODS: Two studies were conducted, each with 6 male participants and 6 female participants, using a stadiometer to accurately measure a person's sitting height. The increase in sitting height after using the True Back II or Rola Stretcher for 10 minutes was compared with that after lying supine for 10 minutes. A third intervention, a firm foam block cut to the same size and shape as the True Back II, was also used in this study. RESULTS: The Rola Stretcher had a significantly greater lengthening effect on the spine compared with supine rest (P <0050). The True Back II had a similar but lesser effect (P <.0509). Women demonstrated a greater height gain than men. CONCLUSION: The True Back II and the Rola Stretcher in particular appear to lengthen the spine after a single use of 10 minutes. The observed discrepancy between the effects in men and women may be an experimental artifact occurring as a result of less effective resetting of the posturing mechanisms in men compared with women. A trend showed the Rola Stretcher to be more effective than the foam block. PMID- 10820299 TI - Are chiropractic tests for the lumbo-pelvic spine reliable and valid? A systematic critical literature review. AB - OBJECTIVE: To systematically review the peer-reviewed literature about the reliability and validity of chiropractic tests used to determine the need for spinal manipulative therapy of the lumbo-pelvic spine, taking into account the quality of the studies. DATA SOURCES: The CHIROLARS database was searched for the years 1976 to 1995 with the following index terms: "chiropractic tests," "chiropractic adjusting technique," "motion palpation," "movement palpation," "leg length," "applied kinesiology," and "sacrooccipital technique." In addition, a manual search was performed at the libraries of the Nordic Institute of Chiropractic and Clinical Biomechanics, Odense, Denmark, and the Anglo-European College of Chiropractic, Bournemouth, United Kingdom. STUDY SELECTION: Studies pertaining to intraexaminer reliability, interexaminer reliability, and/or validity of chiropractic evaluation of the lumbo-pelvic spine were included. DATA EXTRACTION: Data quality were assessed independently by the two reviewers, with a quality score based on predefined methodologic criteria. Results of the studies were then evaluated in relation to quality. DATA SYNTHESIS: None of the tests studied had been sufficiently evaluated in relation to reliability and validity. Only tests for palpation for pain had consistently acceptable results. Motion palpation of the lumbar spine might be valid but showed poor reliability, whereas motion palpation of the sacroiliac joints seemed to be slightly reliable but was not shown to be valid. Measures of leg-length inequality seemed to correlate with radiographic measurements but consensus on method and interpretation is lacking. For the sacrooccipital technique, some evidence favors the validity of the arm fossa test but the rest of the test regimen remains poorly documented. Documentation of applied kinesiology was not available. Palpation for muscle tension, palpation for misalignment, and visual inspection were either undocumented, unreliable, or not valid. CONCLUSION: The detection of the manipulative lesion in the lumbo-pelvic spine depends on valid and reliable tests. Because such tests have not been established, the presence of the manipulative lesion remains hypothetical. Great effort is needed to develop, establish, and enforce valid and reliable test procedures. PMID- 10820300 TI - Lumbar intraspinal synovial cyst containing gas as a cause for low-back pain. AB - OBJECTIVE: To discuss intraspinal synovial cysts caused by degenerative changes involving the posterior articular facets in the lumbar spine and to provide differential considerations for patients with low-back pain. CLINICAL FEATURES: A 70-year-old man with low-back and gluteal pain demonstrating eventual progression of radiating pain into the left thigh, calf, ankle, and foot over a 5-month period. Radiographs of the lumbar spine revealed mild degenerative disk disease at L5-S1 with associated vacuum phenomena of the L5 disk. Degenerative osteophytes were present at L3, L4, and L5. Moderate posterior joint arthrosis was evident at L4-L5 and L5-S1. Computed tomography and magnetic resonance imaging studies revealed an intraspinal gas-containing synovial cyst at the left lateral aspect of the central canal at the level of the left L4-L5 facet articulation. INTERVENTION AND OUTCOME: The patient underwent surgical excision of the synovial cyst with remission of symptoms. CONCLUSION: Gas-containing intraspinal synovial cysts can be a significant finding and a causative factor in patients with low-back pain and pain radiating into the lower extremities. Both computed tomography and magnetic resonance imaging are important in defining intraspinal synovial cysts as a cause of back pain in patients whose low-back pain does not respond to chiropractic care. PMID- 10820301 TI - Case study: acceleration/deceleration injury with angular kyphosis. AB - OBJECTIVE: To discuss the case of a patient who received upper cervical chiropractic care after trauma-induced arcual kyphosis in the cervical spine. A practical application of conservative management for posttrauma cervical spine injury in the private office setting is described. CLINICAL FEATURES: A 17-year old female patient suffered an unstable C3/C4 motor segment after a lateral impact motor vehicle collision. Additional symptoms on presentation included vertigo, tinnitus, neck and shoulder pain, and confusion. INTERVENTION AND OUTCOME: Conservative management consisted exclusively of upper cervical-specific adjustments guided by radiographic analysis and paraspinal bilateral skin temperature differential analysis of the cervical spine. During 10 weeks of care and 22 office visits, all symptoms subsided and the instability of C3/C4 motor segment appeared to be completely resolved. CONCLUSION: This study provides support for the use of upper cervical chiropractic management in cervical spine trauma cases. The clinical work-up consisted of physical examination, radiographic analysis, computer-administered and scored cognitive function testing, and audiometric examination. After conservative care, these examinations were repeated and demonstrated that the objective findings concurred with the subjective improvements reported by the patient. PMID- 10820302 TI - Evidence that tension-type headache and cervicogenic headache are distinct disorders. PMID- 10820303 TI - Presenting chiropractic to medical students. PMID- 10820304 TI - Preliminary study of the effects of a placebo chiropractic treatment with sham adjustments. PMID- 10820305 TI - In response PMID- 10820306 TI - Management of acute lumbar disk herniation initially presenting as mechanical low back pain. PMID- 10820307 TI - In response PMID- 10820308 TI - Chronic spinal pain syndromes: a clinical pilot trial comparing acupuncture, a nonsteroidal anti-inflammatory drug, and spinal manipulation. PMID- 10820309 TI - Possible manifestation of temporomandibular joint dysfunction on chiropractic cervical X-ray studies. PMID- 10820311 TI - In response PMID- 10820310 TI - Torque: an appraisal of misuse of terminology in chiropractic literature and technique. PMID- 10820312 TI - Motor learning and performance: A problem-based learning approach PMID- 10820313 TI - Analysis of nuclear ribonucleoproteic structures during notochordal cell differentiation and maturation in chick embryos. AB - The ultrastructure of notochordal cells and the quantitative changes of nuclear mRNA-containing particles were studied in several stages of the development of the chick embryo. The modifications in the frequency of perichromatin granules (PCG) were analyzed in embryos at 24 hr to 10 days of incubation (stages 6-36 of Hamburger and Hamilton). The ultrastructural and morphometric data show that notochordal cells undergo changes that can be systematized in four periods. Very early notochordal cells (stages 6-11), are characterized by the presence of large nucleoli and abundant PCG, traits probably related to the frequent mitotic division and the expression of inductive signals reported in numerous papers. During the second period (stages 16-21) the number of PCG and the size of the nucleolus decrease. These changes are coincident with the beginning of vacuolization. In the third period (stages 21-30), the notochordal cells undergo a second cytodifferentiation characterized by a large increase of cytoplasmic vacuolization and secretion of materials that thicken the perichordal sheath. During this period, the nucleolus becomes smaller and the number of PCG increases. Similar features were previously described during functional maturation of embryonic neurons and striated fibers at synaptogenesis, and epidermal cells. The fourth period, beginning at stage 30, is characterized by the decrease of the density of PCG and of the nucleolar volume and corresponds to cessation of mitosis and cell degeneration. PMID- 10820314 TI - Periosteally derived osteoblast-like cells differentiate into chondrocytes in suspension culture in agarose. AB - Pluripotent cells from the periosteal layer adjacent to cortical bone attain an osteoblast-like phenotype in culture when reaching confluence in monolayer. It is unknown whether such newly differentiated osteoblast-like cells preserve the chondrogenic potential characteristics for stem cells derived from the periosteum. Primary osteoprogenitor cells derived from bovine metacarpal periosteum were differentiated into alkaline phosphatase-positive osteoblast-like cells by an established monolayer culture protocol. After transfer into suspension culture in agarose gels, the cells differentiated into chondrocytes demonstrated by the production of collagen II, but not of collagen I, as well as alkaline phosphatase activity was abated. Contrarily, with continuation of monolayer culture, the cells maintained their osteoblast-like phenotype and secreted large amounts of collagen I and a minor quantity of collagen III and V. The alkaline phosphatase activity steadily increased during the entire culture period of 2 weeks. Thus, our culture techniques can serve as useful tools to study mechanisms of differentiation by modulating the phenotypic potential of osteogenic cells. The results presented here support the notion that the extracellular environment strongly influences the cell type and its metabolism. PMID- 10820315 TI - Transport and rearrangement of the intra-acrosomal protein acrin1 (MN7) during spermiogenesis in the guinea pig testis. AB - We have recently shown that a 90-kDa glycoprotein, acrin1 (MN7), is exclusively localized in the dorsal region of the acrosomal apical segment of mature guinea pig sperm, and that its location changes during epididymal maturation. The present study examined the process of transport and organization of this protein in the acrosome during spermatogenesis in the guinea pig testis. Immunoperoxidase electron microscopy showed stage-specific localization of acrin1 within the developing acrosome, as follows: acrin1 first appeared in the proacrosomic vesicles of the early Golgi phase spermatids, and it was then localized in the electron-lucent matrix region of the acrosomic vesicles of the late Golgi phase spermatids. During the cap phase, acrin1 was abundant in the electron-lucent matrix of the acrosomal apical segment and in the head-cap region (principal segment). acrin1 became more restricted to the peripheral region of the electron lucent matrix of acrosome phase spermatids and it was localized in the electron lucent dorsal matrix region of maturation phase spermatids. In the final step of spermiogenesis, acrin1 disappeared from the equatorial and principal segments, and it was finally confined to the dorsal matrix region of the acrosomal apical segment. In addition, Western blot analysis showed that acrin1 of testes and epididymal sperm was of the identical size, indicating that acrin1 is not proteolytically modified during epididymal sperm maturation. These results indicate that acrosome morphogenesis is closely associated with the rearrangement of acrosomal proteins. PMID- 10820316 TI - Somatostatin and somatostatin receptors in the pig pineal gland during postnatal development: an immunocytochemical study. AB - An immunohistochemical study of the pineal gland of the domestic pig was carried out using rabbit antisera raised against synthetic peptide fragments corresponding to different amino acid sequences of the prosomatostatin, the somatostatin-14, and the somatostatin-28 molecule. The study was supplemented by immunohistochemical staining with rabbit antisera raised against five subtypes of somatostatin receptors. The pineal glands were taken from the newborn, 21-day-old and 7-month-old pigs. Immunoreactive nerve fibers and cells were observed in the pineal gland with all the antisera against somatostatin and prosomatostatin. The nerve fibers were located throughout the pineal gland-in the capsule, connective septa, and parenchyma-with the highest density in proximo-ventral part of the gland. The somatostatin positive fibers were also found in the habenular and posterior commissurae areas. Somatostatin-immunoreactive cell bodies were observed mostly in the central part of the gland. These results point to the existence of two somatostatin sources in the pig pineal gland: 1) nerve fibers, probably of central origin; and 2) cells that may represent intrapineal neurons or specialised pinealocytes. A clear difference in the immunoreactivity between newborn, 21-day-old, and 7-month-old pigs was found. Generally, the density of nerve fibers was lower in adult than young animals. The number of the cells also decreased with age. By using the antisera against the five somatostatin receptors, only sst3 - receptor immunoreactivity could be detected. The receptor immunoreactivity was confined to varicose and smooth fibers and some cells. The sst(3)-receptor positive structures were localised in all parts of the gland and their number was higher in younger pigs. PMID- 10820317 TI - Development of the anatomical alteration of the cerebellar fissura prima. AB - The development of the naturally occurring malformation of the cerebellar fissura prima was monitored in rats starting from 4 days of life to the adulthood. The first sign of the malformation was evident at 10 days of life and consisted of an interruption of the pia mater and the fusion of the external granular layers on the two sides of the fissura. Later, nests of apparently mature granule cells could be seen to be encircled by cells of the external granular layer and to be connected to the granule cell layer by thin bridges of cells. Calretinin immunoreactive fibers followed the bridges of cells to reach the ectopic masses of cells. Towards the end of histogenesis and in adult animals, brush cells and Golgi cells were present in the ectopic masses of granule cells. The latter appeared to contribute to the formation of normal glomeruli, as in the orthotopic granule cell layer. In addition, bundles of parallel fibers crossed the boundary between the molecular layers on the two side of the fissure, thus suggesting that parallel fibers can contact Purkinje cells of the opposite folium. PMID- 10820318 TI - Mist1 expression is a common link among serous exocrine cells exhibiting regulated exocytosis. AB - Mist1 is a basic helix-loop-helix transcription factor that represses E-box mediated transcription. Previous studies have suggested that the Mist1 gene is expressed in a wide range of tissues, although a complete characterization of Mist1 protein accumulation in the adult organism has not been described. In an effort to identify specific cell types that contain the Mist1 protein, antibodies specific for Mist1 were generated and used in Western blot and immunohistochemical assays. Our studies show that the Mist1 protein is present in many different tissues but that it is restricted to cell types that are exclusively secretory in nature. Pancreatic acinar cells, serous or seromucous cells of the salivary glands, chief cells of the stomach, and secretory cells of the prostate and seminal vesicle show high levels of Mist1 protein, whereas nonserous exocrine cells, including the mucus-producing cells of the salivary glands, remain Mist1 negative. These results identify Mist1 as the first transcription factor that exhibits this unique serous-specific expression pattern and suggest that Mist1 may have a key role in establishing and maintaining a pathway responsible for the exocytosis of serous secretions. PMID- 10820319 TI - MMP-2 expression during early avian cardiac and neural crest morphogenesis. AB - Matrix metalloproteinase-type 2 (MMP-2) degrades extracellular matrix, mediates cell migration and tissue remodeling, and is implicated in mediating neural crest (NC) and cardiac development. However, there is little information regarding the expression and distribution of MMP-2 during cardiogenesis and NC morphogenesis. To elucidate the role of MMP-2, we performed a comprehensive study on the temporal and spatial distribution of MMP-2 mRNA and protein during critical stages of early avian NC and cardiac development. We found that ectodermally derived NC cells did not express MMP-2 mRNA during their initial formation and early emigration but encountered MMP-2 protein in basement membranes deposited by mesodermal cells. While NC cells did not synthesize MMP-2 mRNA early in migration, MMP-2 expression was seen in NC cells within the cranial paraxial and pharyngeal arch mesenchyme at later stages but was never detected in NC-derived neural structures. This suggested NC MMP-2 expression was temporally and spatially dependent on tissue interactions or differed within the various NC subpopulations. MMP-2 was first expressed within cardiogenic splanchnic mesoderm before and during the formation of the early heart tube, at sites of active pharyngeal arch and cardiac remodeling, and during cardiac cushion cell migration. Collectively, these results support the postulate that MMP-2 has an important functional role in early cardiogenesis, NC cell and cardiac cushion migration, and remodeling of the pharyngeal arches and cardiac heart tube. PMID- 10820320 TI - Nonadrenergic innervation of the rat laryngeal vasculature. AB - In order to gain a better understanding of the central and local control of laryngeal blood flow, the vascular innervation to the rat laryngeal muscles was examined. To visualize the vascular network, the animals were perfused with a gelatin/India ink solution. The larynges were removed and fixed. The superior laryngeal, cricothyroid, and inferior laryngeal arteries (all branch off the superior thyroid artery) were dissected in continuity into their respective muscles. Specimens were reacted in toto using immunohistochemical techniques for the presence of neuropeptide-Y (NPY), vasoactive intestinal peptide (VIP), calcitonin gene related peptide (CGRP), and neuronal nitric oxide synthase (NOS 1). Results show that all of the laryngeal vasculature is richly innervated by fibers containing these peptides. Qualitatively, the most prominent of these is NPY in association with the superior and the inferior laryngeal arteries, followed by VIP and NOS-1, and finally CGRP distributed equally on all the vessels. Immunopositive fibers are found along the entire course of the feeding arteries, beginning with the superior thyroid artery and continuing down to small arterioles into the terminal vascular beds. These peptides can act as vasodilators, vasoconstrictors, and/or neuromodulators and may work synergistically or antagonistically with other transmitters in controlling laryngeal blood flow. Their effects are dependent on the specific vascular bed in question, that is, in some areas they are vasodilators, in others vasoconstrictors, and in other neuromodulators. What effects they have on the laryngeal vasculature and how they interact within the larynx have yet to be determined. PMID- 10820321 TI - Observations on the muscles of the eye of the bowhead whale, Balaena mysticetus. AB - The muscles of the eyelids and the extraocular muscles of mysticete whales are poorly described for a variety of reasons, including considerable difficulty in obtaining specimens. Our objective is to provide such a description for the bowhead whale, Balaena mysticetus. This study has examined the gross anatomy of the region in six specimens (five adults, one fetus) of the bowhead whale. Results show that the muscles associated with the eye are well developed and possess several distinctive features. For example, precise limits of each extraocular muscle are difficult to determine along their entire length because these muscles intermingle with one another near their insertion. Furthermore, some fibers from these muscles (except the retractor bulbi) also insert into the eyelids. Pulling on these muscles to simulate contraction results in movement of the eyelids and suggests a role for these muscles in palpebral retraction. Insertion of a large levator palpebrae superioris muscle into the upper eyelid further enhances opening of the palpebral fissure. Another unusual feature is the presence of tunnel-like structures that redirect the dorsal and ventral oblique muscles. The dorsal oblique muscle is redirected caudally about 90 degrees, then directed medially by another 90 degrees. These directional changes are accomplished via a connective tissue tunnel derived in part from the fibrous connective tissue of the dorsal rectus and the levator palpebrae superioris muscles. In most terrestrial mammals, a similar change in direction is accomplished by a cartilaginous trochlea. The ventral oblique muscle originates via a slender tendon from the frontal bone and undergoes a similar radical change in direction. Its tendon of insertion undergoes about a 90-degree change in direction that is accomplished through a tunnel-like structure derived from fibrous connective tissue of the ventral rectus muscle. Based on the morphology of the musculature presented, it is likely that the eyeballs and eyelids of the bowhead whale are quite mobile and appear capable of complex movement. The possibility of retraction and protrusion of the eyeball is discussed. PMID- 10820322 TI - Closer look at lactose-mediated support of retinal morphogenesis. AB - We have previously shown in intact isolated eye rudiments from Xenopus laevis that lactose, but not mannose, permits the formation of organized photoreceptor outer segments in the absence of the retinal pigment epithelium (RPE). The purpose of this study was to determine, using electron microscopic analysis, the key ultrastructural differences between healthy retinas, lactose-protected retinas, and retinas that developed aberrantly to reveal which subcellular structures were exclusively present in healthy retinas. Filamentous actin was also localized in retinas to determine its distribution under the various conditions. In healthy retinas, calycal processes extending approximately three fourths of the length of the outer segment surrounded highly organized photoreceptor outer segments. Adherens junctions were localized between adjacent photoreceptors and Muller cells at the outer limiting membrane. In addition, Muller cells possessed apical processes that extended for a short distance beyond the adherens junctions. These fine cytoarchitectural details were missing in retinas that completed differentiation in the absence of the RPE; both calycal and apical processes were no longer present and adherens junctions were sparsely intermittent. Muller cells appeared atrophic. Similarly, mannose promoted none of the fine cytoarchitectural details of the retina. Lactose, however, supported the formation of the proper subcellular cytoarchitecture of both photoreceptor and Muller cells. These results suggest that these subcellular structures may be fundamental for the proper assembly and stability of organized outer segments and are necessary to allow for normal cytogenesis of the outer retina. PMID- 10820323 TI - Immunocytochemical characteristics of cells and fibers in the nasal mucosa of young and adult macaques. AB - The mammalian nasal cavity is lined by an olfactory mucosa (OM) and a respiratory mucosa (RM). The principal OM cell type is the olfactory receptor neuron (ORN). However, little is known about ORNs in the life histories of primates. The RM, similar to the RM in the tracheobronchial tract (TBT), is dominated by ciliated columnar cells. Neuroendocrine cells (NECs) are essential in the TBT; little is known about nasal NECs. This study examined the immunolabeling characteristics of primate OM and RM for three important proteins-calretinin (CR), olfactory marker protein (OMP), and protein gene product 9.5 (PGP). Tissues from newborn to 15 year-old macaques were analyzed to determine the expression of these proteins during various stages of development. Standard immunocytochemistry on aldehyde fixed tissues was applied, utilizing the avidin-biotin peroxidase (ABC) method. Immuno-electron microscopy confirmed the immunoreactive cell types. ORNs were immunoreactive for CR, OMP, and PGP at all ages studied. Immunoreactivity for PGP also was displayed in a subset of ciliated, columnar epithelial cells in the RM and in an extensive network of subepithelial fibers spread throughout both mucosae. The results suggest that macaque ORNs express three important proteins over a wide life history, and that the macaque may be a reliable model for studying primate/human olfaction during aging. The PGP-labeling results also suggest that the macaque nasal peptidergic fibers express PGP and that the respiratory epithelium contains NECs with labeling characteristics similar to those in the TBT. PMID- 10820324 TI - High resolution imaging of the mouse inner ear by microtomography: a new tool in inner ear research. AB - A newly developed desktop microtomograph was used to evaluate whether it is suitable for visualizing the three-dimensional (3D) morphology of the mouse inner ear (at a micrometer level) and whether it is applicable as a fast screening tool to detect hereditary abnormalities in this organ. To this end, the epistatic circler, a mutant mouse showing abnormal circling behaviour, was used as a model. The inner ears were dissected out, formaldehyde-fixed, and scanned at maximal resolution along the longitudinal axis. After segmentation, stacks of tomographic images were used for 3D reconstruction of the bony labyrinth. Finally, the obtained data were correlated with subsequent conventional histological examination. The spatial resolution (8 microm) achieved by this instrument, was found to be far superior to that obtained by conventional computer tomography (CT) and magnetic resonance (MR)-imaging equipment. The technique provides detailed tomographic images of the bony labyrinths and enables an adequate 3D reconstruction of the inner ear structures in this small mammal. In addition, it allows a screening for pathologic specimens prior to the more time- and labour consuming histological techniques, which are still essential to gather information at a (sub)cellular level. This imaging technique can be regarded as a valuable tool in future research on hereditary inner ear abnormalities. PMID- 10820325 TI - Laboratory scaledown of protein purification processes involving fractional precipitation and centrifugal recovery. AB - The ability to predict the performance of large-scale processes is central to the rapid development of successful operations at the pilot and industrial scale. In this article, we examine the operation, at laboratory scale, of precipitation reactors and centrifuges for protein precipitate recovery and dewatering and how they might best mimic large-scale reactors and centrifuges, in this case, a pilot scale batch stirred-tank reactor and a multichamber-bowl centrifuge. Novel approaches to bench-top centrifuge operation are provided, in particular with a view to delivery of material for subsequent high-resolution purification, which would be obtained at full pilot scale. Results are presented in terms of properties of the protein precipitates, the fraction of solids recovered, and the extent of dewatering achieved. Good agreement was obtained at bench scale (a 1000 fold scale down factor) for all of these parameters for pilot-scale, batch-feed operation. In addition, the methodology developed allows identification of the extent of break-up that occurs in continuous-feed centrifuges when processing shear-sensitive materials such as the protein precipitates studied here. PMID- 10820326 TI - Different expression of an S-adenosylmethionine synthetase gene in transgenic tobacco callus modifies alkaloid biosynthesis. AB - Transformed callus cultures of Nicotiana tabacum were generated in which the SAM 1 gene from Arabidopsis thaliana encoding S-adenosylmethionine synthetase (SAM S), under the control of the 35S promoter, had been integrated. The presence of the SAM-1 gene was detected in all tested transformants and the SAM-S activity correlated with the accumulation of SAM in the tobacco callus cultures. Three distinct phenotypic classes were identified among the transgenic cell lines in relation to growth of the cells, structure of the calli, and level of SAM. Transgene silencing was observed in several cultivated transgenic calli and this phenomenon was correlated directly with a low level of SAM-1 mRNA accompanied by a decrease of the SAM-S activity. The transgenic calli overexpressing the SAM-1 gene accumulated a high SAM level. The modifications in SAM-S activity were reflected in the pattern of secondary products present in the different cell lines, thereby demonstrating that the flux through the biosynthetic pathway of a plant secondary product can be modified by means of genetic engineering. PMID- 10820327 TI - A model for high-pressure ultrafiltration of blood. AB - A semi-analytic model to predict the permeate flux during high-pressure ultrafiltration of blood with highly permeable membranes is proposed. This model explicitly considers the hydraulic resistance of the retained particles that limits the flux. An empirically derived relationship between particle surface concentration and hydraulic resistance is used. This model incorporates the axial variations in blood cell and solute surface concentrations (or concentration polarization), shear-induced diffusion coefficient for the blood cells, effective diffusion coefficient for the blood solutes, hydraulic (lumen) pressure, and flow rate. This model agrees well with experimental results in the pressure independent filtration flux region. PMID- 10820328 TI - Dynamic kinetic resolution of suprofen thioester via coupled trioctylamine and lipase catalysis. AB - A lipase-catalyzed enantioselective hydrolysis process under conditions of continuous in situ racemization of substrate with trioctylamine as the catalyst was developed for the production of (S)-suprofen from (R,S)-suprofen 2,2,2 trifluoroethyl thioester in isooctane. A detailed investigation of trioctylamine concentration on the enzyme activation and stability as well as the kinetic behaviors of the thioester in racemization and enzymatic reaction was conducted, in which good agreement between the experimental data and theoretical results was observed. A complete conversion of the racemate for the desired (S)-suprofen in 95% ee(P) was obtained. Moreover, the recovery of the acid product by extraction and reuse of the organic solution were reported. PMID- 10820329 TI - Characterization of polymeric buffers for operating membrane-trapped enzyme reactors in an electric field. AB - A novel class of amphoteric, polymeric buffers, is described, consisting of grafting onto growing polyacrylamide chains weakly acidic and basic acrylamido monomers (called Immobilines; protolytic groups as N-substituents on the nitrogen of the amido bond), for operating a membrane-immobilized enzyme reactor (MIER) in an electric field. With these soluble, polymeric buffers, it is possible to operate the membrane reactor at any optimum of pH activity, for any given enzyme, in the pH 3-10 scale. Such buffers, being amphoteric, are confined in the enzyme reaction chamber by the same isoelectric trapping mechanism. The best buffers were found to be those polymerized in presence of 9% neutral monomer (acrylamide) and containing 20 mM Immobiline as buffering ion. To decrease their viscosity in solution, the polymeric buffers are synthesized at high temperatures (70 degrees C) and in presence of a chain-transfer agent. The weight average molecular size in these conditions has been found to be ca. 200,000 Da. These buffers exhibited excellent performance in a variety of enzyme reactions in the MIER, such as in the case of penicillin G acylase and histidine decarboxylase and were found to greatly stabilize enzyme activity, permitting operation of the MIER over extended periods of time. As an example, in a penicillin G acylase reactor, >75% enzyme activity was maintained over a 10-d cycle of operation, while with conventional buffers more than 90% inactivation was experienced over the same period of time. This novel class of macromolecular, amphoteric buffers could also be exploited in other types of conventional bioreactors not based on an isoelectric trapping mechanism. PMID- 10820330 TI - Biomass evolution in porous media and its effects on permeability under starvation conditions. AB - The purpose of this study was to understand bacteria profile modification and its applications in subsurface biological operations such as biobarrier formation, in situ bioremediation, and microbial-enhanced oil recovery. Biomass accumulation and evolution in porous media were investigated both experimentally and theoretically. To study both nutrient-rich and carbon-source-depleted conditions, Leuconostoc mesenteroides was chosen because of its rapid growth rate and exopolymer production rate. Porous micromodels were used to study the effects of biomass evolution on the permeability of a porous medium. Bacterial starvation was initiated by switching the feed from a nutrient solution to a buffer solution in order to examine biofilm stability under nutrient-poor conditions. Four different evolution patterns were identified during the nutrient-rich and nutrient-depleted conditions used in the micromodel experiments. In phase I, the permeability of the porous micromodel decreased as a result of biomass accumulation in pore bodies and pore throats. In phase II, starvation conditions were initiated. The depletion of nutrient in the phase II resulted in slower growth of the biofilm causing the permeability to reach a minimum as all the remaining nutrients were consumed. In phase III, permeability began to increase due to biofilm sloughing caused by shear stress. In phase IV, shear stress remained below the critical shear stress for sloughing and the biofilm remained stable for long periods of time during starvation. The critical shear stress for biofilm sloughing provided an indication of biofilm strength. Shear removal of biofilms occurred when shear stress exceeded critical shear stress. A network model was used to describe the biofilm formation phenomenon and the existence of a critical shear stress. Simulations were in qualitative agreement with the experimental results, and demonstrate the existence of a critical shear stress. PMID- 10820331 TI - Continuous enzymatic production of peptide precursor in aqueous/organic biphasic medium. AB - N-(benzyloxycarbonyl)-L-aspartic acid (Z-L-Asp) has generally been used as a carboxyl substrate for the enzymatic synthesis of a precursor of aspartame (synthetic sweetener); however, alternative inexpensive protection groups have been in demand for lowering the total cost of its industrial-scale production. A formyl group (F-) was found to be a more desirable protecting group for the N terminus of amino acid derivatives due to its low cost of preparation, introduction, and removal. The yield of F-AspPheOMe (N-formyl-L-aspartyl-L-phe- nylalanine methylester), however, was found to be <10% in a conventional aqueous medium. We found that F-L-Asp and L-PheOMe were partitioned mainly to the aqueous phase in an aqueous/organic biphasic medium, whereas F-AspPheOMe partitioned to the organic phase, especially when some extracting agents were added. In this study, simultaneous operation of an enzymatic reaction and a product separation by liquid-liquid extraction was thus applied to the F-AspPheOMe synthesis. We succeeded in synthesizing F-AspPheOMe continuously in an aqueous/tributylphosphate (TBP) biphasic medium with >95% yield, which was about tenfold higher than that in an aqueous monophasic medium. PMID- 10820332 TI - Reactive extraction of penicillin G in hollow-fiber and hollow-fiber fabric modules. AB - Penicillin G (Pen G) can be rapidly extracted in hollow-fiber liquid-liquid contactors using N-lauryl-N-trialkylmethylamine (Amberlite LA-2) as the extractant. n-Butylacetate is much better than decanol as a diluent for such an extraction, although decanol can give a partition coefficient four times larger. The overall mass transfer coefficient found is a function of aqueous flow on the lumen side of the fiber, and is less dependent on shell-side flow. In backextraction, the overall mass transfer coefficient determined is only one tenth that of the forward extraction, primarily because the hydrophobic hollow fibers used have a high mass transfer resistance under these conditions. The mass transfer experiments show that hollow-fiber extraction of Pen G is competitive with centrifugal extraction. The prospects for extraction of other fermentation products with hollow fibers can be estimated based on the present study. PMID- 10820333 TI - Achievement of high cell density and high antibody productivity by a controlled fed perfusion bioreactor process. AB - Controlled feeding of nutrient supplements to a cell culture to enhance monoclonal antibody productivity has been practiced widely in high-yield, fed batch processes. In this study, a similar feeding concept has been applied to a perfused culture and evaluated for the effects on bioreactor productivity and product quality. Our experimental results show that, by using such a "controlled fed perfusion" approach, the volumetric antibody productivity (antibody per liter per day) was significantly increased by nearly twofold over the perfusion process, and surpassed fed-batch and batch processes by almost tenfold. The substantial boost in the overall productivity is attributable primarily to the combined effects of increased cell density as well as reduced product dilution. Both were achieved through careful nutrient supplementation in conjunction with metabolite minimization. As the manufacturing process evolved from roller bottles to the controlled-fed perfusion bioreactor system, the immunoreactivity and the cDNA sequences of the antibody were well preserved. However, the product glycosylation distribution patterns did alter. The controlled-feed perfusion process demonstrated a unique encompassment of the advantages of fed-batch and perfusion methods; that is, high product concentration with high volume throughput. Therefore, it may be very suitable for large-scale production of monoclonal antibodies. PMID- 10820334 TI - Aqueous two-phase systems containing urea: influence of protein structure on protein partitioning. AB - During recombinant E. coli fermentation with high-expression levels inclusion bodies are often formed. Aqueous two-phase systems have been successfully used in the presence of urea for the initial recovery step of inclusion bodies from E. coli. Basic studies of the complex interactions are lacking. For a systematic study of protein partitioning in the presence of urea we selected T4-lysozyme mutants with different thermal stability as a model. The stabilization of these variants by phase components was investigated measuring the fluorescence emission of tryptophan residues in the protein. Protein structure was stabilized at pH 7 in the order of S0(4)(2-) >> PEG = Dextran > H(2)O. The conformation of proteins was shown to have a strong influence on the partitioning in aqueous two-phase systems. Tryptophan and its homologuous di- and tripeptdides were partitioned in similar phase systems to normalize for contribution from hydrophobic interactions. PMID- 10820335 TI - Production of enantiopure styrene oxide by recombinant Escherichia coli synthesizing a two-component styrene monooxygenase. AB - A whole cell biocatalytic process was developed to enable the efficient oxidation of styrene to chiral (S)-styrene oxide with an enantiomeric excess better than 99%. Recombinant Escherichia coli cells were employed to express the genes styAB encoding the styrene monooxygenase of Pseudomonas sp. strain VLB120 from an expression plasmid utilizing the alk regulatory system of P. oleovorans GPo1. The strains reached specific activities of up to 70 U* (g cell dry weight)(-1) in shake-flask experiments with glucose as the carbon source. An efficient two liquid phase fed-batch process was established for the production of (S)-styrene oxide with hexadecane as an apolar carrier solvent and a nutrient feed consisting of glucose, magnesium sulfate, and yeast extract. Engineering of the phase fraction and the composition of organic phase and feed led to a 2-L scale process with maximal volumetric productivities of 2.2 g (S)-styrene oxide per liter liquid volume per hour. This optimized process was based completely on defined medium and used bis(2-ethylhexyl)phthalate as the apolar carrier solvent, which together with substrate and inducer consisted of 50% of the total liquid volume. Using this system, we were able to produce per liter liquid volume 11 g of enantiopure (S)-styrene oxide in 10 h. PMID- 10820336 TI - Purification of plasmid DNA by tangential flow filtration. AB - A simple, scalable method for purification of plasmid DNA is described. The method includes modification of the classical alkaline-lysis-based plasmid extraction method by extending the solubilization step from less than 30 min to 24 h. The extraction is followed by the novel use of tangential flow filtration (TFF) for purification of the remaining contaminants. The method does not include the use of any organic solvents, RNase, high-speed centrifugation, or column chromatography steps. The method typically yields 15 to 20 mg of plasmid DNA per liter of bacterial culture and results in removal of >99% of RNA and >95% of the protein that remains after the modified alkaline lysis procedure. The procedure has been demonstrated to be effective in the isolation of seven different plasmids. Plasmids isolated using this method had comparable transfection capability relative to plasmid isolated using a classical, cesium chloride gradient-based method. PMID- 10820337 TI - Transformation of mono- and dichlorinated phenoxybenzoates by phenoxybenzoate dioxygenase in pseudomonas pseudoalcaligenes POB310 and a modified diarylether metabolizing bacterium. AB - Pseudomonas pseudoalcaligenes POB310 contains genes that encode phenoxybenzoate dioxygenase. The enzyme transforms mono- and dichlorinated phenoxybenzoates to yield protocatechuate that is used as a growth substrate and chlorophenols that are nonmetabolizable. Mass spectral analysis of (18)O metabolites obtained from the protocatechuate 3,4-dioxygenase-deficient mutant, POB310-B1, suggested that the reaction mechanism is a regioselective angular dioxygenation. A cloning vector containing reaction relevant genes (pD30.9) was transferred into Pseudomonas sp. strain B13 containing a modified ortho-cleavage pathway for aromatic compounds. The resultant Pseudomonas sp. strain B13-D5 (pD30.9) completely metabolized 3-(4-chlorophenoxy)benzoate. During growth on 3 phenoxybenzoate, strain B13-D5 (pD30.9) (K(s) = 0.70+/-0.04 mM, mu(max) = 0.45+/ 0.03 h(-1), t(d) = 1.5 h, Y = 0.45+/-0.03 g bio- mass x g substrate(-1)) was better adapted to low substrate concentrations, had a faster rate of growth, and a greater yield than POB310 (K(s) = 1.13+/-0.06 mM, mu(max) = 0.31+/-0.02 h(-1), t(d) = 2.2 h, Y = 0.39+/-0.02 g biomass. g substrate(-1)). PMID- 10820338 TI - Increase of hybrids in yeast electrofusion by mitochondrial mutations. AB - Yeast mitochondria were found to affect the zeta-potential of protoplasts, resulting in electrofusion of membrane behavior. For modeling purposes, two fusion systems were investigated: (1) parent yeasts of Saccharomyces cerevisiae G706 (rho(+)) x O708-11-16A (rho(+)), with zeta-potentials of -10 to -27 mV and 28 to -45 mV, depending on MgCl(2) concentration in the medium, respectively; and (2) parent yeasts of S. cerevisiae G706-1 (rho(-)) x O708-11-16A (rho(+)), with zeta-potentials of -30 to -60 mV, depending on MgCl(2). Yields of the hybrids in system (2) were over 100-fold higher than those in system (1). Thus, regulation of yeast electrofusion was found to be possible by mitochondrial mutations. PMID- 10820339 TI - Exposure. PMID- 10820340 TI - Increased expression of matrix metalloproteinase 7 and 9 and membrane type 1 matrix metalloproteinase in esophageal squamous cell carcinomas. AB - BACKGROUND: The 5-year survival rate of patients with submucosal (sm) esophageal carcinoma continues to be much worse than that with intramucosal (m) lesions. The reasons are unclear, but matrix metalloproteinases (MMPs) and membrane type matrix metalloproteinases (MT-MMPs) are known to be involved in degradation of extracellular matrix macromolecules associated with tumor cell invasion. Expression of these enzymes may have some relation to the difference in survival between patients with sm and m esophageal carcinomas. METHODS: Surgical specimens from 148 primary esophageal squamous cell carcinomas were examined for expression of MMP-1, 2, 3, 7, and 9; tissue inhibitor of MMP (TIMP)-1 and 2; MT1-MMP; and MT2-MMP by immunohistochemical staining. The results were compared with tumor progression and other clinicopathologic parameters. Localization of gelatinase activity was also confirmed in carcinoma cells by in situ zymography. RESULTS: Expression of MMP-7 and 9, expression of MT1-MMP, and gelatinase activity were elevated in the carcinomas, correlating with depth of tumor invasion. Percentages of positive sm cases were 44.0%, 51.9%, and 52.0%, respectively, significantly higher than for the m cases. Percentages of venous invasion positive cases were 61.9%, 65.1%, and 65.1%, respectively. CONCLUSIONS: Expression of MMP-7 and 9 and MT1-MMP is closely associated with invasion depth and venous invasion in esophageal squamous cell carcinomas. PMID- 10820341 TI - A clinicopathologic study of gastric mucosa-associated lymphoid tissue lymphoma. AB - BACKGROUND: It is still unclear which patients with gastric mucosa-associated lymphoid tissue (MALT) lymphoma will benefit from the eradication of Helicobacter pylori. METHODS: The authors studied a total of 34 patients. Twenty-three patients had primary gastric lymphoma and underwent gastric resection as initial treatment. Eleven patients with gastric MALT lymphoma who received antibiotics against H. pylori as initial treatment were also included. In all 34 patients, the presence of H. pylori, endoscopic findings, and pathologic features were evaluated. Immunohistochemical expression of Bcl-2, p53, and proliferating cell nuclear antigen (PCNA) was classified as follows: (-), no reactive cells; (+), scattered positive cells; (2+), nests of positive cells; (3+), diffuse positive cells. RESULTS: Patients with low grade MALT lymphoma (LG) tended to be positive for H. pylori (6 of 9), to localize within the submucosa (7 of 9), not to have lymph node involvement (7 of 8), and to have lower tumor stage compared with patients with high grade MALT components (HG). Bcl-2 protein was expressed with high frequency by LG (7 of 9). Strong expression of p 53 was more common in the HG tumors (4 of 14), and strong expression of PCNA showed a significant difference between LG (1 of 8) and HG patients (12 of 13). Investigation of the patients with long term follow-up (n = 4) revealed that LG remained superficial for a long time and showed gradual progression. Most of these tumors were Bcl 2+/p53-approximately+/-/ PCNA- approximately +. There were two patients whose superficial LG (sm/Bcl-2+/p53-/PCNA- approximately +) regressed after the disappearance of H. pylori. On the other hand, one patient developed ulcerated LG (sm/Bcl-2 /p53+/PCNA3+) after disappearance of H. pylori. The authors found complete regression of MALT lymphoma in 9 of 11 patients after H. pylori eradication. Initial tumors of these 9 patients were superficial/sm/n(-)/low grade/Bcl-2+approximately +/-/p53-approximately+ (n = 9), /PCNA-approximately+(n = 6), /PCNA 2+ (n = 3). Two local recurrence and one non-Hodgkin lymphoma in other sites were observed after initial therapy. CONCLUSIONS: Gastric MALT lymphoma with (H. pylori positive/superficial/sm/low grade/Bcl-2 +/p53- approximately +/PCNA- approximately +) pattern will disappear after a patient is cured of H. pylori infection. PMID- 10820342 TI - The role of thymidine phosphorylase expression in the invasiveness of gastric carcinoma. AB - BACKGROUND: Thymidine phosphorylase (TP) has angiogenic activity in various cancer tissues. Gastric carcinomas are classified into two histologic groups: differentiated and undifferentiated adenocarcinomas. There are differences in the modes of development and the extent of infiltration between the two groups. The purpose of the current study was to determine whether TP is involved in the invasiveness and progression of these two types of gastric carcinoma. METHODS: To investigate the expression and localization of TP and the microvessel counts, the authors examined specimens from 149 gastric carcinoma patients. The specimens were stained using monoclonal antibody against TP and polyclonal antibody against factor VIII. To determine the cell type expressing TP, immunohistochemical staining using a monoclonal antibody against CD68 that is specific for macrophages and double staining using antibodies to both TP and CD68 were performed. RESULTS: The proportion of TP positive tumors in differentiated adenocarcinomas was higher than that in undifferentiated adenocarcinomas. The TP positive differentiated adenocarcinomas invaded more deeply than the TP negative ones, but this was not the case with undifferentiated adenocarcinomas. TP was expressed mainly in the invasive edges of tumors and was expressed more frequently in macrophages than in tumor cells. TP expression was correlated with microvessel count and CD68 expression. Patients with TP positive carcinomas had a poorer prognosis than those with TP negative differentiated adenocarcinomas. CONCLUSIONS: TP expressed in macrophages may be correlated with microvessel count and play an important role in tumor invasiveness and progression in differentiated gastric adenocarcinoma. PMID- 10820343 TI - Pericolonic tumor deposits in patients with T3N+MO colon adenocarcinomas: markers of reduced disease free survival and intra-abdominal metastases and their implications for TNM classification. AB - BACKGROUND: A pericolonic tumor deposit (PTD) is a grossly palpated adenocarcinomas within pericolonic adipose tissue not within a lymph node. The source and prognostic significance of PTDs has not been well defined. METHODS: The authors studied 418 T3N+M0 colon adenocarcinomas to determine the frequency and significance of PTDs. They also step-sectioned 30 PTDs to determine their origin and assist in their optimum TNM classification. RESULTS: Seventy-one (18%) of 400 consecutively examined cases had PTDs. The actuarial 1-, 2-, and 5-year disease free survival rates were significantly lower among patients with a PTD. PTDs, regardless of size, significantly impacted disease free survival. Increasing numbers of PTDs was associated with shorter disease free survival. Adenocarcinoma grade, a PTD, increasing numbers of PTDs, and number of lymph node metastases were independently associated with shorter disease free survival. The likelihood of extrahepatic abdominal failure was proportionally greater with increasing numbers of PTDs. Adenocarcinoma was observed in perineural, peri-large vessel, or intravascular locations in step-sectioned PTDs. CONCLUSIONS: A PTD is a perineural, perivascular, or intravascular tumor extension beyond the muscularis propria. They are distinct from lymph node metastases and should not be considered their prognostic equivalent. The disease free survival impact of even small PTDs was significant, suggesting that PTDs of all sizes should be considered a single entity. TNM classification of PTDs as lymph node metastases or discontinuous tumor extension is probably not accurate. The number and greatest dimension of PTDs should be reported separately from lymph node metastases. PMID- 10820344 TI - High expression of vascular endothelial growth factor is associated with liver metastasis and a poor prognosis for patients with ductal pancreatic adenocarcinoma. AB - BACKGROUND: Vascular endothelial growth factor (VEGF), a recently identified growth factor with significant angiogenic properties, is a multifunctional angiogenic cytokine that is expressed in many tumors. High VEGF expression has been shown to correlate with the incidence of metastasis and poor prognosis in various cancers. In this study, the authors investigated VEGF expression and microvessel density (MVD) in ductal pancreatic adenocarcinoma and examined the correlations among VEGF expression, clinicopathologic factors, and clinical outcome. The authors especially focused on the correlation between VEGF expression and liver metastasis. METHODS: Paraffin embedded tumor specimens of 142 surgically resected pancreas carcinoma were immunohistochemically stained for VEGF and MVD. The correlations among VEGF expression and MVD, clinicopathologic factors, and clinical outcome were then statistically analyzed. RESULTS: One hundred thirty-two (93%) of 142 ductal pancreatic adenocarcinomas were positive for VEGF protein by immunohistochemistry. A significant correlation was observed between VEGF positivity and MVD (P < 0.0001). Multivariate logistic regression analysis indicated a significant association between high VEGF expression and liver metastasis (P = 0.010) but no other factors, such as age, tumor size, histologic type, lymph node metastasis, venous invasion, neural invasion, peritoneal metastasis, or local recurrence. Patients with tumors that showed moderate or high VEGF expression had significantly shorter survival than patients with low VEGF expression or none at all in their tumors (P < 0.05). CONCLUSIONS: These results indicate that VEGF expression is closely correlated with MVD and seems to be an important predictor for both liver metastasis and poor prognosis in ductal pancreatic adenocarcinoma. PMID- 10820345 TI - Elective ipsilateral neck irradiation of patients with locally advanced maxillary sinus carcinoma. AB - BACKGROUND: The current study was conducted to investigate retrospectively whether elective ipsilateral neck irradiation (EINI) is effective in controlling subclinical neck disease in patients with locally advanced (T3 and T4) nonmetastatic (N0, M0) squamous cell carcinoma (SCC) of the maxilla. METHODS: Between 1987 and 1993 a total of 44 patients were treated with EINI. The primary tumor bed was treated with 60 grays (Gy) in patients undergoing radical maxillectomy or with 66 Gy in patients undergoing partial maxillectomy. The ipsilateral upper and lower neck (down to the clavicle) was treated with either opposing anteroposterior-posteroanterior fields or appositional electron fields. The dose of elective neck radiotherapy was 50 Gy in 25 daily fractions. RESULTS: The 5-year and 10-year survival rates (with standard error [SE]) were 66% (SE 7%) and 60% (SE 8%), respectively, whereas the 5-year and 10-year recurrence free survival rates both were 64% (SE 7%). The 10-year local recurrence free survival rate was 69% (SE 7%), whereas the 10-year regional recurrence free survival rate was 94% (SE 4%). Of the 2 patients who developed a recurrence in the neck, 1 was salvaged successfully by surgery, producing an ultimate 10-year regional recurrence free survival rate of 97%. The 10-year distant metastasis free survival rate was 91% (SE 4%). CONCLUSIONS: The findings of the current study appear to suggest the potential efficacy of EINI in patients with locally advanced, nonmetastatic SCC of the maxilla and that the high rate of control of cervical lymph nodes may lead to better overall survival than that reported in the majority of the recent series. PMID- 10820346 TI - Prognostic significance of an increased number of micrometastatic tumor cells in the bone marrow of patients with first recurrence of breast carcinoma. AB - BACKGROUND: Using cytokeratin (CK) as a histogenetic marker of epithelial tumor cells in the bone marrow of patients with primary breast carcinoma, a subgroup of patients with decreased survival can be identified. This study was designed to evaluate the frequency and prognostic relevance of such cells in patients with recurrent breast carcinoma. METHODS: Bone marrow aspirates from 65 patients were analyzed immunocytochemically for the presence of CK positive cells. A quantitative immunoassay with monoclonal anti-CK antibody A45-B/B3 was used and 2 x 10(6) bone marrow cells per patient were evaluated. For prognostic evaluation the authors calculated a cutoff value of micrometastatic tumor cells by analogy to classification and regression tree (CART) analysis. Patients were monitored prospectively for a median of 37 months (range, 11-63 months). RESULTS: Bone marrow micrometastases were present in 5 of 32 patients (16%) with locoregional recurrence and in 24 of 33 patients (73%) with distant recurrence. The bone marrow status yielded no prognostic indication for patients with locoregional recurrence. In contrast, a cutoff value of 2.5 tumor cells per 1 million bone marrow cells analyzed (2.5 x 10(-6) tumor cells) correlated with a significantly different prognosis for women with distant disease. Patients with metastatic disease and a micrometastatic tumor load of > 2.5 x 10(-6) tumor cells survived for a mean of 6 months (95% confidence interval [95% CI], 2.0-9.1) compared with 17 months (95% CI, 11.6-22.0) for patients with < or = 2.5 x 10(-6) tumor cells (P < 0.0001). Multivariate analysis, allowing for hormone receptor status, disease free interval prior to recurrence, manifestation site of metastases, age, and micrometastases in bone marrow, revealed that bone marrow involvement was an independent risk factor, with a hazard ratio of 7.4 (95% CI, 1.6-13.3) for disease-related death. CONCLUSIONS: An increased number of micrometastases identified in the bone marrow of patients with metastatic breast carcinoma represents an independent prognostic factor that may influence future therapeutic strategies for patients with metastatic breast carcinoma. PMID- 10820347 TI - The effects of radiation therapy on quality of life of women with breast carcinoma: results of a randomized trial. Ontario Clinical Oncology Group. AB - BACKGROUND: The purpose of this study was to evaluate the effect of breast irradiation on quality of life, including cosmetic outcome, for patients enrolled in a clinical trial. METHODS: Between 1984 and 1989, a randomized trial was conducted in Ontario, Canada, in which women with lymph node negative breast carcinoma who had undergone lumpectomy and axillary lymph node dissection were randomized to either breast irradiation or no further treatment. A modified version of the Breast Cancer Chemotherapy Questionnaire (BCQ) was administered to women at baseline, 1 month (4 weeks), and 2 months (8 weeks) after randomization. Irritation of the skin of the breast, breast pain, and appearance of the breast to the patient were also assessed every 3 months for the first 2 years of the study. RESULTS: Of 837 patients, 416 were randomly allocated to radiation therapy and 421 to no further treatment. The mean change in quality of life from baseline to 2 months was -0.05 for the radiation group and +0.30 for the control group. The difference between groups was statistically significant (P = 0.0001). Longer term radiation therapy increased the proportion of patients who were troubled by irritation of the skin of the breast and breast pain. Radiation therapy did not increase the proportion of patients at 2 years who were troubled by the appearance of the treated breast; 4.8% in irradiated and nonirradiated patients (P = 0.62). CONCLUSIONS: Breast irradiation therapy had an effect on quality of life during treatment. After treatment, irradiated patients reported increased breast symptoms compared with controls. However, no difference was detected between groups at 2 years in the rates of skin irritation, breast pain, and being upset by the appearance of the breast. PMID- 10820348 TI - Early cervical carcinoma: the natural history of lymph node involvement redefined on the basis of thorough parametrectomy and giant section study. AB - BACKGROUND: Although parametrectomy is the most difficult step in the surgical treatment of cervical carcinoma and is the main cause of postoperative complications, little attention has been given to the patterns of parametrial spread. METHODS: Sixty-nine patients with previously untreated cervical carcinoma (Federation Internationale de Gynecologie et d'Obstetrique [FIGO] Stage IB1, 49 patients [71%]; Stage IB2, 8 patients [12%]; and Stage IIA, 12 patients [17%]; squamous, 59 patients [86%]; and adenocarcinoma, 10 patients [14%]) underwent radical hysterectomy and pelvic +/- aortic lymphadenectomy. Hysterectomy specimens were processed with the giant section technique. To obtain a thorough three-dimensional assessment of the paracervical tissue, both the superficial and deep layers of the cervicovesical ligament (anterior parametrium) and the uterosacral ligament (posterior parametrium) were separated from the uterus and submitted for pathologic evaluation. After resection of the lateral parametrium with hemoclips, the lympho-fatty tissue remaining around the pudendal vessels was removed carefully and referred to as "the distal part of the lateral parametrium." RESULTS: When analyzing all the parametria, lymph nodes were present in 64 patients (93%). Clinically undetected parametrial involvement was found by pathologic examination in 15 Stage IB1 patients (31%), 5 Stage IB2 patients (63%), and 7 Stage IIA patients (58%). Metastases were found in the cardinal, cervicovesical, and sacrouterine ligaments and principally were comprised of lymph node and vascular space invasion. Twenty-five patients (36%) had pelvic lymph node metastases whereas concomitant parametrial involvement was observed in all patients. The overall 5-year survival was 91%, being higher for parametria and lymph node negative patients (100%) than for those with lymph node and/or parametrial metastases (78%). CONCLUSIONS: A three-dimensional pathologic assessment showed that subclinical parametrial spreading of the so-called "early" tumors (Stage IB-IIA) occurred in approximately 30-60% of these patients, and metastasis to the pelvic lymph nodes always was associated with parametrial disease. A better understanding of the patterns of parametrial diffusion will improve knowledge of the natural history of cervical carcinoma and in the future may influence the treatment of these patients. Furthermore, pathologic assessment of cervical carcinoma should be modified to evaluate correctly the parametrial status of each patient. The current routine pathologic evaluation of the parametria makes it very difficult to detect lymph node metastases and tumor emboli. PMID- 10820349 TI - Photodetection of cervical intraepithelial neoplasia using 5-aminolevulinic acid induced porphyrin fluorescence. AB - BACKGROUND: Screening for cervical carcinoma and its precursors is based on cervical cytology and diagnostic colposcopy. Despite the decrease in the incidence of cervical carcinoma in countries with a good screening program, this rate of decline is leveling off. Known problems are false-negative rates of cytology and low specificity of colposcopy. This clinical study examined the diagnostic potential of porphyrin fluorescence in patients with cervical intraepithelial neoplasia Grade 1-3 (CIN 1-3). METHODS: Sixty-eight women attending our colposcopy clinic underwent a gynecologic examination, including cytology, human papillomavirus (HPV) testing, and colposcopy. They received 10 mL 0.5% or 1.0% 5-aminolevulinic acid (5-ALA) topically. After 30-360 minutes, real time image analysis was performed, and spectra were obtained from 685 sites. RESULTS: Due to rapid photobleaching, 0.5% 5-ALA proved ineffective for fluorescence assessment. Using 1% 5-ALA, the authors found that fluorescence intensities correlated with incubation time; however, fluorescence contrast showed a maximum at 60-90 minutes (ratio 11:1). HPV DNA positive lesions showed significantly higher fluorescence. Fluorescence imaging after 60-90 minutes achieved similar sensitivity and specificity compared with colposcopy in detecting CIN with 94% and 51% versus 95% and 50%, respectively. However, the specificity was markedly improved by fluorescence spectroscopy, achieving 75%. The evaluation of spectral measurements revealed significantly higher values for CIN compared with normal tissue and for CIN 2/3 compared with CIN 1 (P < 0.001). CONCLUSIONS: Using a time interval of 60-90 minutes after topical application of 1% 5-ALA, the authors observed specific porphyrin fluorescence of CIN. Fluorescence spectroscopy promises to become a valuable tool for the diagnosis of CIN. PMID- 10820350 TI - Papanicolaou smear history and diagnosis of invasive cervical carcinoma among members of a large prepaid health plan. AB - BACKGROUND: Despite the widespread use of Papanicolaou (Pap) smear screening, substantial morbidity and mortality from cervical carcinoma continue in the U.S. Although access to screening is a major barrier to use of the Pap smear, invasive cervical carcinoma (ICC) still is observed in health plan members who have comprehensive preventive care coverage. METHODS: For all women diagnosed with ICC between 1988 and 1994 in a large prepaid health plan, the authors retrospectively reviewed the medical records for prediagnosis Pap smear history to identify antecedents to ICC. RESULTS: Of 642 women diagnosed as having ICC, 455 (71%) had been plan members for > or = 30 of the 36 months before diagnosis. Of these 455 women, 240 (53%) had no Pap smear during the 6-36 months prior to diagnosis (i.e., were nonadherent to screening), 127 (28%) had only "normal" Pap smear results, 42 (9%) had at least 1 abnormal Pap smear and were adequately followed, 17 (4%) had at least 1 abnormal result without adequate follow-up, and 29 (6%) were classified as "other." Compared with adherent women, more nonadherent women presented with later stage disease, were symptomatic at the time of diagnosis, were older, and were of a race/ethnicity other than non-Hispanic white. CONCLUSIONS: Nonadherence to screening recommendations was found to be the most important modifiable antecedent to ICC in this population. The rate of incidence of ICC could be reduced by interventions to increase screening in women who do not have Pap smears regularly and by the use of newer screening technologies to reduce the false-negative rate of Pap smears. PMID- 10820351 TI - Frameshift mutations at coding mononucleotide repeat microsatellites in endometrial carcinoma with microsatellite instability. AB - BACKGROUND: Microsatellite instability (MI) is a frequent occurrence in endometrioid carcinoma of the endometrium (EC). Several genes known to contain mononucleotide short tracts in their coding sequences (TGF-beta RII, IGFIIR, BAX, hMSH6, and hMSH3) are likely targets for mutations in these tumors. METHODS: DNA from 24 patients with EC and MI was extracted from blood and from fresh-frozen and paraffin embedded tumor tissue. Seven of these patients were found to have metastatic spread to paraaortic lymph nodes. DNA also was studied from 10 patients with EC without MI. RESULTS: Frameshift mutations at coding mononucleotide repeats were detected by single strand conformation polymorphism analysis and DNA sequencing. Frameshift mutations were detected more frequently in BAX (11 of 24 MI positive (+) tumors; 45.8%) than in TGF-beta RII (0 of 24 tumors; 0%), IGFIIR (3 of 24 tumors; 12.5%), hMSH3 (6 of 24 tumors; 25%), or hMSH6 (0 of 24 tumors; 0%). The mutations frequently were distributed heterogeneously throughout the tumors. Overall, frameshift mutations at 1 or more of these mononucleotide repeat microsatellites were found in 17 of 24 MI+ tumors (70.8%) but in none of the 10 MI negative neoplasms. In the seven EC patients with lymph node metastases, mutations in IGFRII were found more commonly in those with metastatic (three of seven patients) rather than primary (one of seven) tumors. CONCLUSIONS: The results of the current study confirm that BAX is an important target gene in ECs with MI. The frequent detection of IGFRII frameshift mutations in lymph node metastases suggest that IGFRII may play a role in tumor progression in these patients. PMID- 10820352 TI - Transvaginal color Doppler ultrasonic characterization of benign and malignant ovarian cystic teratomas and comparison with serum squamous cell carcinoma antigen. AB - BACKGROUND: The preoperative diagnosis of squamous cell carcinoma (SCC) arising in mature cystic teratoma of the ovary remains difficult. The purpose of this study is to examine the usefulness of transvaginal color Doppler ultrasound (TV CDU) in differentiating malignant (SCC) from benign cystic teratoma of the ovary. METHODS: Eighty-eight patients with an ovarian tumor showing gray scale sonographic appearances of mature cystic teratoma were preoperatively evaluated for the presence or absence of intratumoral blood flow by TV-CDU. The blood flow characteristics of the tumor vessels were analyzed using the resistance index (RI), pulsatility index (PI), and peak systolic velocity (PSV). The serum levels of SCC antigen were also randomly examined preoperatively in 50 patients. RESULTS: Intratumoral blood flow was significantly detected in malignant teratomas (SCCs) (80.0%; 4 of 5) compared with benign teratomas (20.5%; 17 of 83) (P < 0.01). All malignant teratomas with intratumoral blood flow showed both RI less than 0.4 and PI less than 0.6, whereas no benign teratomas showed any such value except for 1 case with struma ovarii. In addition, both the mean RI and the mean PI values in the tumor vessels were significantly lower in the malignant teratomas (RI: 0.31 +/- 0.07; PI: 0.40 +/- 0.16) than in the benign teratomas (RI: 0.62 +/- 0.13; PI: 1.06 +/- 0.44) (P < 0.001). However, the mean PSV value of the malignant teratomas (PSV: 20.6 +/- 8.33) was not significantly different from the benign teratomas (PSV: 18.1 +/- 9.9). Elevation of serum SCC was found in 4 of 5 patients (80%) with malignant teratomas, whereas the elevation was found in 11 of 45 patients (24.4%) with benign teratomas (P < 0.05). The diagnostic accuracy using the RI (cutoff value 0.4) as well as the PI (cutoff value 0.6) was thus 95.2%, which was significantly superior to that obtained by using the serum SCC (76%) (cutoff value, 1.5 ng/mL). CONCLUSIONS: Evaluating the presence or absence of intratumoral blood flow, together with blood flow resistance, in tumor vessels using TV-CDU thus may be more useful to differentiate malignant (SCC) from benign cystic teratomas of the ovary than by measuring serum SCC levels. PMID- 10820353 TI - The correlation of serial prostate specific antigen measurements with clinical outcome after external beam radiation therapy of patients for prostate carcinoma. AB - BACKGROUND: The authors analyzed retrospectively their institution's experience in treating patients with localized prostate carcinoma with external beam radiation therapy (EBRT) to determine the correlation of various biochemical failure (BF) definitions with clinical failure and cause specific survival (CSS). METHODS: Between January 1987 and December 1997, 1,094 patients with clinical T1 T3N0M0 prostate carcinoma were treated with definitive EBRT alone at William Beaumont Hospital, Royal Oak, Michigan. All patients received EBRT alone (no adjuvant hormones) to a median total prostate dose of 66.6 grays (Gy) (range, 59.4-70.4 Gy). Multiple BF definitions were tested for their correlation with clinical failure and cause specific death (CSD = 1-CSS). All BF definitions were tested for sensitivity, specificity, and accuracy of predicting subsequent clinical failure and CSD. Positive and negative predictive values were calculated in the form of 10-year actuarial clinical failure and CSD rates. Analyses were performed on all 1,094 patients as well as for those 727 patients who had at least 5 post-RT prostate specific antigen (PSA) level measurements and who did not receive hormonal therapy for post-RT PSA elevations. The median PSA follow-up was 4.0 years for the entire population and 4.5 years for those 727 patients included in the second analysis. RESULTS: In the entire population, 167 patients (15%) experienced clinical failure corresponding to 5- and 10-year actuarial rates of 16% and 34%, respectively. The correlation of various BF definitions with outcome was calculated in those 727 patients who did not receive hormonal therapy. For these patients, BF (as defined by the American Society for Therapeutic Radiology and Oncology Consensus Panel) yielded a 73% sensitivity, 76% specificity, and 75% overall accuracy for predicting clinical failure and a 74% sensitivity, 69% specificity, and 69% overall accuracy for predicting CSD. The 10-year clinical failure rate for those 251 patients demonstrating 3 consecutive PSA rises (BF) was 64% versus 14% for those patients who did not meet these criteria (biochemically controlled [BC]). As expected, definitions requiring only two rises were more sensitive but less specific in predicting clinical failure than those definitions requiring three or four rises. Because there were dramatically more clinically controlled patients (85%) than clinical failures (15%), the overall accuracy for each definition more closely approximated its specificity. The definitions classifying BF as a postnadir increase of > or = 3 or > or = 4 ng/mL above the nadir yielded the highest accuracies of 87% and 88%, respectively. In addition, these definitions also appeared to provide the greatest separation in clinical failure rates between BC and BF patients, an absolute difference of 77% and 76%, respectively. CONCLUSIONS: The correlation between BF and clinical failure and CSD varies markedly depending on the BF definition used. Definitions incorporating a fixed baseline (the nadir level) and the postnadir PSA profile may have better correlation with clinical failure than definitions using the nadir only or a specific number of consecutive rises in which a variable baseline "resets" after a PSA decrease. PMID- 10820354 TI - Father-son testicular tumors: evidence for genetic anticipation? A case report and review of the literature. AB - BACKGROUND: The etiology of testicular carcinomas is unknown. Father-son testicular tumors represent an important subset of patients because the sons have a 6- to 10-fold increase in the risk of developing a testicular tumor compared with the general population, suggesting a possible genetic component in the etiology of the disease. Genetic anticipation has been established for several diseases and manifests itself clinically with earlier age of onset (AO) of disease or with increased severity of disease in children compared with their parents. METHODS: We report only the 13th case of pure seminoma occurring in both a father and a son. The most striking feature of our case study was a 27-year difference in the AO between father and son. A review of the literature from 1972 through 1999 provided 47 cases of father-son testicular tumors in which sufficient information was available to compare the AO and the severity of the disease in the fathers (G1) and the sons (G2). RESULTS: The mean AO for G1 was 43.3 (+/- 1.6) years compared with a mean AO for G2 of 27.0 (+/- 1.0) years for all 47 cases (P < 0.01). For fathers presenting with a pure seminoma, the mean AO was 45.9 (+/- 1.8) years for G1 compared with 27.0 (+/- 1.1) years for G2 (P < 0.01). For fathers presenting with a nonseminomatous tumor, the mean AO in G1 was 37.9 (+/- 2.6) years and 26.9 (+/- 1.8) years for G2 (P < 0.01). For the entire group of 47 cases, disease was more severe in G2 compared with G1 in 43% of cases, of equal severity in 47% of cases, and less severe in G2 compared with G1 in 10% of cases. CONCLUSIONS: Genetic anticipation may be responsible for many father-son testicular tumors. PMID- 10820355 TI - Predicting the survival of bladder carcinoma patients treated with radical cystectomy. AB - BACKGROUND: Clinical outcomes vary for patients treated with radical cystectomy. The authors sought to identify factors associated with the survival of patients treated with radical cystectomy for urothelial carcinoma of the urinary bladder. METHODS: The authors studied 218 patients treated with radical cystectomy for urothelial carcinoma between 1980 to 1984. Patient ages ranged from 41 to 78 years (mean, 64 years). Using the 1997 TNM system, T classifications were Ta (17 patients), T1 (44), T2 (71), T3a (42), T3b (14), T4a (28), and T4b (2). Thirty two patients had lymph node metastasis at the time of surgery. Histologic grade was determined according to the newly proposed World Health Organization and International Society of Urological Pathology grading system; tumor was low grade in 43 patients and high grade in 175. The male-to-female ratio was 4.9 to 1. The mean follow-up of patients still alive was 13.1 years (median, 13.8 years; range, 30 days to 18 years). Cox proportional hazards models were used to determine the impact of numerous clinical and pathologic findings on survival. RESULTS: Ten year local recurrence free, distant metastasis free, cancer specific, and all cause survival were 71%, 73%, 67%, and 41%, respectively. In univariate analysis, cancer size, T classification, and lymph node status were associated with distant metastasis free, cancer specific, and all-cause survival. Histologic grade and surgical margin status were significantly associated with worse cancer specific and all-cause survival, but not with distant metastasis free survival. In multivariate analysis, cancer size, margin status, T classification, and lymph node status were identified as significantly associated with cancer specific survival after adjustment for age and gender. CONCLUSIONS: Long term survival is achieved in a significant number of patients treated with radical cystectomy. In this study, patients with organ-confined (< or = pT2) and small size (< or = 3 cm) cancer had favorable 10-year distant metastasis free (93%) and cancer specific survival (88%) after cystectomy. Tumor size, margin status, extravesical involvement, and lymph node metastasis are important pathologic factors and should be considered as stratification variables in identifying patients for whom adjuvant chemotherapy should be evaluated in clinical trials. PMID- 10820357 TI - Discrepancies in diagnoses of neuroepithelial neoplasms: the San Francisco Bay Area Adult Glioma Study. AB - BACKGROUND: Valid and reliable diagnoses of disease are key both to meaningful epidemiologic and clinical investigations and to decision-making about appropriate treatment. One previous study highlighted the lack of precision in diagnosing primary brain tumors in a neuropathology referral practice. The current study explores diagnostic discrepancies in a population-based adult glioma series by hospital of origin, specialty training of the original diagnosing pathologist, and clinical significance. METHODS: To confirm patients' eligibility for the San Francisco Adult Glioma Study, the authors obtained participants' pathology specimens and conducted a uniform secondary neuropathology review. Eligible patients were all adults age 20 years or older newly diagnosed with glioma between August 1, 1991, and March 31, 1994, who resided in 1 of 6 San Francisco Bay Area counties. RESULTS: Overall, the original and secondary diagnoses were the same (concordant) for 352 (77%) of the 457 cases available for study. Twenty-six percent of the cases from community hospitals were discordant, compared with 12% of the cases from academic hospitals P= 0.004. Of the 105 discordant diagnoses, 17 (16%) were determined to be clinically significant, defined as a difference that could significantly alter patient management and/or prognosis. Sixteen of these 17 cases originated at community hospitals, and only 1 originated at a hospital with a neuropathologist. Based on the distribution of review diagnoses, subjects presenting at nonacademic hospitals were more likely than those presenting at academic hospitals to have glioblastoma (61% vs. 52%; P = 0.07). CONCLUSIONS: The percentage of cases with discrepant original and review diagnoses was higher among those originally diagnosed at community hospitals without a neuropathologist than among those originally diagnosed at an academic hospital with a neuropathologist. Clinically significant discrepancies were much more likely to have originated at a community hospital without a neuropathologist. These data highlight the importance of review of brain tumors by a neuropathologist prior to decision-making regarding treatment. A separate implication of this study is that glioma cases selected exclusively from academic or nonacademic institutions in a particular geographic area are unlikely to be representative of all cases occurring in that area. PMID- 10820356 TI - Expression of the human mismatch repair gene hMSH2: a potential marker for urothelial malignancy. AB - BACKGROUND: The human mismatch repair (MMR) gene hMSH2 (human mutS homolog-2) is a DNA repair gene that has been reported to be mutated in 40% of hereditary nonpolyposis colon cancer (HNPCC) kindreds and a small percentage of sporadic tumors. HNPCC is a cancer predisposition syndrome with an increased risk of carcinoma of the colon, endometrium, stomach, small intestine, ovary, ureter, and renal pelvis. Immunohistochemical analysis demonstrated increased hMSH2 expression in sporadic colon carcinoma and in the replicative compartment of normal epithelium. A recent immunohistochemical analysis of hMSH2 in bladder tumors correlated reduced hMSH2 expression with recurrence and higher tumor grade. In the current study, we examined hMSH2 expression in urothelial malignancy using immunohistochemical analysis and developed a molecular assay for the detection of hMSH2 expression in bladder washes. METHODS: Immunohistochemical analysis of 17 tumors from the genitourinary tract and reverse transcription coupled with polymerase chain reaction (RT-PCR) of 40 bladder washes were used to investigate hMSH2 expression in noninvasive and invasive urothelial malignancies. RESULTS: Increased expression of hMSH2 was detected in all tumors examined using immunohistochemical analysis independent of grade or stage. Reverse transcription PCR of hMSH2 mRNA from bladder washes detected 17 of 21 patients with primary or recurrent urothelial neoplasms or tumors involving the urothelial system. Four patients with urothelial malignancies without detectable hMSH2 expression from their bladder washes had high grade lesions. Ten of 13 patients without pathologic or cystoscopic evidence of bladder tumors were negative for hMSH2 expression in bladder washes. Two patients with bladder tumors and bladder washes that were positive for hMSH2 subsequently were found to be negative for hMSH2 after treatment of their tumors and at last follow-up had remained recurrence free for at least 1 year. CONCLUSIONS: The results of the current study suggest that hMSH2 expression is increased in low and high grade urothelial neoplasms, similar to the expression pattern in sporadic colon carcinoma. However, a fraction of high grade lesions may not express hMSH2 as detected by RT-PCR from bladder washes. The ability to detect hMSH2 expression in bladder washes may allow the use of hMSH2 expression as a marker for urothelial malignancy. In addition, the ability to define hMSH2 deficient tumors using bladder washes may have prognostic significance in the treatment of patients with urothelial carcinoma. PMID- 10820358 TI - Therapy for patients with high grade astrocytoma using intraarterial chemotherapy and radiation therapy. AB - BACKGROUND: High grade astrocytomas account for approximately 40% of all primary brain tumors. The median survival is approximately 8-10 months for patients with glioblastoma multiforme and 36 months for patients with anaplastic astrocytoma. The results of systemic chemotherapy in the treatment of brain tumors have been reported to be less than satisfactory, mainly because of the blood-brain barrier impermeability for chemotherapeutic drugs. Intraarterial chemotherapy has been an attractive alternative with which to overcome this problem. METHODS: Eighty-three patients with high grade astrocytoma (glioblastoma multiforme [63 patients] and anaplastic astrocytoma--[20 patients]) were treated with intraarterial (intracarotid and/or intravertebral) chemotherapy and radiation therapy between 1987 and 1997. Patients received cisplatin, 60 mg/m2, and etoposide, 40 mg/m2. Radiation therapy was delivered either after completion of the chemotherapy or concomitantly with the chemotherapy. RESULTS: Thirty-four of 71 evaluable patients with high grade astrocytoma (48%) responded to the chemotherapy. The median survival for patients with glioblastoma multiforme who received chemotherapy prior to radiation therapy was 20 months versus 7 months for those patients who underwent concomitant chemotherapy/radiation therapy. Patients with anaplastic astrocytoma who received chemotherapy prior to radiation therapy had a median survival of 45 months compared with 12 months for patients who received concomitant chemotherapy/ radiation therapy. The toxicity profile has been reported to be mild and well tolerated. CONCLUSIONS: Intraarterial chemotherapy for patients with glioblastoma multiforme, delivered prior to radiation therapy, appears to result in a median survival three times longer than that achieved with concomitant chemotherapy/radiation therapy. In addition, patients appear to survive substantially longer than they do after radiation therapy with the addition of systemic chemotherapy. Side effects are reported to be acceptable. PMID- 10820359 TI - The incidence of lymphoma in first-degree relatives of patients with Hodgkin disease and non-Hodgkin lymphoma: results and limitations of a registry-linked study. AB - BACKGROUND: The precise incidence of familial Hodgkin disease (HD) and non Hodgkin lymphoma (NHL) in first-degree relatives is unknown. Through record linkage using two population-based sources, the authors estimated the risk of HD and NHL in family members of lymphoma probands. METHODS: The authors identified 8,037 first-degree relatives of 2,606 lymphoma cases (28.5% HD, 71.5% NHL) treated between 1970 and 1993 in 3 hospitals in Israel via the family file of the Population Registry. The authors linked this file with the Israel Cancer Registry, then calculated the standardized incidence ratio (SIR) by dividing the observed number of cases with the expected, adjusting for age, gender, calendar year, and continent of origin. RESULTS: The family file yielded incomplete ascertainment of relatives (for 771 probands, no relatives were identified). Twenty cases of lymphoma--6 HD and 14 NHL--were identified among relatives of lymphoma patients. The SIR for HD was 1.15 (95% confidence interval [CI]: 0.42 2.51) and for NHL 1.71 (95% CI: 0.93-2.87), considering the entire population of first-degree relatives. SIRs among siblings of lymphoma probands were 3.12 (95% CI: 1.01-7.29) for HD, 2.16 (95% CI: 0.45-6.31) for NHL, and 2.68 (95% CI: 1.15 5.27) for all lymphomas. There were 4 HD/HD, 1 NHL/NHL, and 3 NHL/HD sibling pairs. For HD/HD and NHL/NHL sibling pairs, the interval between lymphoma occurrence in proband and sibling was 1-4 years, whereas for HD/NHL pairs this ranged from 16 to 21 years. CONCLUSIONS: The risk of lymphoma among siblings of lymphoma probands was over 2.5-fold that of the general population and lower among other family members. The temporal proximity of HD/HD and NHL/NHL sibling pairs argues for environmental as well as genetic etiology. This method was hampered by incomplete data. PMID- 10820360 TI - Onycholysis as a complication of systemic chemotherapy: report of five cases associated with prolonged weekly paclitaxel therapy and review of the literature. AB - BACKGROUND: Onycholysis has been reported in association with the use of several noncytotoxic drugs and with chemotherapy in 135 patients. Onycholysis may be precipitated by exposure to ultraviolet radiation. METHODS: The authors studied 91 patients who received paclitaxel and 187 patients who received doxorubicin. RESULTS: Onycholysis occurred in 5 of 21 patients who received > 6 courses of weekly paclitaxel, developing in the summer months in all 5 patients. It did not occur in patients who received fewer weekly paclitaxel courses or those who were treated every 3 weeks. Onycholysis did not occur in 187 patients who received doxorubicin. Review of the literature revealed that onycholysis is nearly exclusively associated with anthracycline and taxane therapy. CONCLUSIONS: Prolonged weekly paclitaxel, other taxanes, and anthracyclines cause onycholysis in some patients, which may be precipitated by exposure to sunlight. Patients receiving these drugs should protect their nails from sunlight. PMID- 10820361 TI - Genomic imprinting of insulin-like growth factor-2 in infant leukemia and childhood neuroblastoma. AB - BACKGROUND: Loss of imprinting (LOI) of insulin-like growth factor-2 (IGF-2) has been implicated in the pathogenesis of certain human cancers and tumor predisposing overgrowth disorders, such as Beckwith-Wiedemann syndrome. In a previous study, the authors revealed that certain patients with childhood acute leukemia and neuroblastoma had had rapid somatic growth after birth, suggesting the involvement of growth factor(s) in tumorigenesis. In the current study, the authors examined whether relaxation of IGF-2 imprinting occurred in infant leukemia and childhood neuroblastoma. METHODS: The genomic DNA of infant leukemia, childhood neuroblastoma, and control individuals was amplified by polymerase chain reaction (PCR). Patients who had heterozygous genotype were selected as informative cases using Apa I polymorphism in exon 9 of the IGF-2 gene. Total RNA was isolated from informative cases, followed by cDNA synthesis. cDNA was amplified by PCR, and direct sequence was performed for determining allele specific transcription. RESULTS: Twenty of 22 infant leukemia blasts and all of 16 neuroblastoma cells showed normal monoallelic expression of IGF-2 as well as 23 controls. The height and weight of two acute lymphoblastic leukemia patients with LOI were within normal ranges for Japanese children. CONCLUSIONS: The current study revealed that the imprinting status of IGF-2 was generally maintained in infant leukemia and confirmed that it was maintained in childhood neuroblastoma. The results suggest that LOI of IGF-2 does not play a major role in the carcinogenesis of these diseases or in rapid physical growth of the patients. PMID- 10820362 TI - T-cell-rich large B-cell lymphoma in children and adolescents: a clinicopathologic report of six cases from the Children's Cancer Group Study CCG 5961. AB - BACKGROUND: T-cell-rich large B-cell lymphoma (TCRLBCL) is a morphologic subset of diffuse large B-cell lymphoma that has been confused with Hodgkin disease and reactive lymphadenopathies. To the authors' knowledge the majority of reports of TCRLBCL are from adults, and it is not widely recognized as occurring in the pediatric population. The current study reports a cohort of six cases of TCRLBCL from the Children's Cancer Group CCG-5961 study. METHODS: Biopsies from patients entered on CCG-5961 were submitted for central pathology review and immunophenotyping. Six cases of TCRLBCL were identified and correlated with clinical characteristics. RESULTS: Of 86 cases centrally reviewed to date on CCG 5961, 20 (23%) were diagnosed as diffuse large B-cell lymphomas. Of these, 6 cases (7% of total cases and 30% of large B-cell cases) were TCRLBCL, based on a diffuse growth pattern with a minor population of neoplastic large B cells and an associated extensive reactive T-cell infiltrate. All patients with TCRLBCL were males ages 12-16 years. Three patients with TCRLBCL had advanced stage disease. No bone marrow or central nervous system involvement was detected in any case. CONCLUSIONS: TCRLBCL is a morphologic subtype of diffuse large B-cell lymphoma that may be difficult to recognize due to the extensive infiltrate of reactive T cells. This entity is not well recognized in pediatric patients, but in the current study represented 7% of all cases and 30% of large B-cell lymphomas received for central review from the ongoing CCG-5961 protocol. Because TCRLBCL may be confused with Hodgkin disease and reactive lymphadenopathies, it is essential that this entity be recognized in the pediatric age group. PMID- 10820363 TI - A brief pain diary for ambulatory patients with advanced cancer: acceptability and validity. AB - BACKGROUND: Undertreatment of pain is common among ambulatory patients with advanced cancer. Available pain assessment tools are complex and not easy to use in ambulatory care settings. METHODS: The authors developed and assessed the acceptability and psychometric properties of a simple, brief 4-week pain diary for ambulatory care assessing 3 indicators: current pain intensity on rising and retiring, number of daily rescue doses, and weekly impact of pain on quality of life. Selected European Organization for Research and Treatment (EORTC) Quality of Life Questionnaire subscales were administered once concurrently for validation purposes. Subjects were 98 adult French-speaking ambulatory patients with advanced cancers who were on opioids, free of apparent cognitive impairment, and recruited through 2 oncology clinics in Quebec City, Quebec, Canada. RESULTS: In the first 3 weeks, > or =80% of patients completed all requested diary data. Internal consistencies of the 5-item scale assessing pain impact on quality of life were 0.87-0.92 over the study period. Pain intensity predicted both increased use of rescue doses and negative pain impact on quality of life. This latter scale was also responsive to decreases in pain intensity over 1-week intervals. As hypothesized, pain intensity, rescue doses, and pain impact on quality of life correlated with EORTC scale scores. Pain intensity correlated most strongly with the EORTC pain and global quality of life scales (r = 0.65 and -0.55, respectively). CONCLUSIONS: This diary is simple, very brief, acceptable to patients, and appears to be valid. It can thus likely be used to monitor pain management for advanced cancer patients. PMID- 10820364 TI - The annual report to the nation on the status of cancer, 1973-1997, with a special section on colorectal cancer. AB - BACKGROUND: This annual report to the nation addresses progress in cancer prevention and control in the U.S. with a special section on colorectal cancer. This report is the joint effort of the American Cancer Society, the National Cancer Institute (NCI), the North American Association of Central Cancer Registries (NAACCR), and the Centers for Disease Control and Prevention (CDC), including the National Center for Health Statistics (NCHS). METHODS: Age-adjusted rates were based on cancer incidence data from the NCI and NAACCR and underlying cause of death as compiled by NCHS. Joinpoint analysis was based on NCI Surveillance, Epidemiology, and End Results (SEER) program incidence rates and NCHS death rates for 1973-1997. The prevalence of screening examinations for colorectal cancer was obtained from the CDC's Behavioral Risk Factor Surveillance System and the NCHS's National Health Interview Survey. RESULTS: Between 1990 1997, overall cancer incidence and death rates declined. Joinpoint analyses of cancer incidence and death rates confirmed the declines described in earlier reports. The incidence trends for colorectal cancer have shown recent steep declines for whites in contrast to a leveling off of the rates for blacks. State to-state variations occurred in colorectal cancer screening prevalence as well as incidence and death rates. CONCLUSIONS: The continuing declines in overall cancer incidence and death rates are encouraging. However, a few of the top ten incidence or mortality cancer sites continued to increase or remained level. For many cancer sites, whites had lower incidence and mortality rates than blacks but higher rates than Hispanics, Asian and Pacific Islanders, and American Indians/Alaska Natives. The variations in colorectal cancer incidence and death rates by race/ethnicity, gender, age, and geographic area may be related to differences in risk factors, demographic characteristics, screening, and medical practice. New efforts currently are underway to increase awareness of screening benefits and treatment for colorectal cancer. PMID- 10820365 TI - 5-fluorouracil-induced small bowel toxicity in patients with colorectal carcinoma. PMID- 10820366 TI - Author reply. PMID- 10820367 TI - Author reply. PMID- 10820368 TI - Classification of gastric carcinoma using the Goseki system provides prognostic information additional to TNM staging. PMID- 10820369 TI - Author reply. PMID- 10820370 TI - Interaction within clusters of dendritic cells and helper T cells during initial Th1/Th2 commitment. AB - Cytokines are the main agents known to regulate Th1 / Th2 commitment, where they may operate through paracrine activity within clusters of T cells gathered around dendritic cells (DC). An in vitro system is used here to test this possibility, using clusters around DC composed of naive TCR-transgenic ovalbumin peptide 323 - 339-specific CD4(+) T cells as targets plus TCR-transgenic pigeon cytochrome C peptide 88 - 104-specific CD4(+) polarized Th1 or Th2 cells as inducers. The polarized inducer cells exerted their maximum effect when the two T cell populations were activated within the same cluster, implemented by allowing a single DC to present both their epitopes. This finding thus supports the paracrine hypothesis. The system was then employed to explore the role of individual cytokines by means of inhibition by monoclonal antibodies. Development of Th2 commitment proved strictly dependent on the IL-4 produced by the Th2 inducers. For Th1 commitment, IFN-gamma and IL-12 were both needed, but with IFN gamma required only during the initial period of culture. The rapid timing observed under these conditions places constraints on the molecular basis of commitment, and appears accurately to reflect the physiological response in vivo. PMID- 10820371 TI - Early myeloid cells are high producers of nitric oxide upon CD40 plus IFN-gamma stimulation through a mechanism dependent on endogenous TNF-alpha and IL-1alpha. AB - Bone marrow contains nonadherent low-density wheat germ agglutinin-positive (Fr3 WGA(+)) cells that release large amounts of NO and show natural suppressor activity if stimulated with activated T cells. We have assessed the involvement of CD40-derived signals in NO production and their cytokine requirements. Production of NO by Fr3-WGA(+) cells in co-culture with activated T cells is inhibited by a competing CD40 soluble fusion protein. Fr3-WGA(+) cells express the inducible NO synthase (iNOS) and release NO following CD40 plus IFN-gamma activation. Production of NO through CD40 is strictly dependent on endogenous TNF alpha and / or IL-1alpha, since it is inhibited by neutralizing these cytokines or blocking the TNF receptor (p55). Both cytokines are transcribed when Fr3 WGA(+) cells are stimulated by CD40 signaling plus IFN-gamma, although TNF-alpha remains below detection limits in stimulated Fr3-WGA(+) cell cultures. Phenotypic studies combined with data on intracellular iNOS expression and cell sorting indicate that NO-producing cells are CD40, CD31 (ER-MP12), CD11b (Mac-1)low, ER MP20 (Ly-6C) and Gr-1 (Ly-6G) positive, consistent with myeloid progenitors. The results point to early myeloid cells as an important cell source of NO once triggered by activated T cells through CD40 and IFN-gamma-derived signals, in a mechanism involving the production of TNF-alpha and / or IL-1alpha. PMID- 10820372 TI - Cell desensitization by sublytic C5b-9 complexes and calcium ionophores depends on activation of protein kinase C. AB - Basal cell resistance to lysis by complement C5b-9 complexes depends on extracellular and intracellular protection. Cell membrane regulatory proteins and enzymes interfere with complement activation and intracellular processes of protein phosphorylation and synthesis support cell resistance and damage repair. K562 human erythroleukemic cells treated with sublytic complement doses become protected from lytic doses of complement within 50 min. The early signaling processes leading to cell desensitization to complement-mediated lysis were studied. Treatment with calcium ionophores or phorbol 12-myristate 13-acetate rapidly induced in K562 cells protection from complement as well as synthesis of a large protein complex similar to the large complement-induced protein complex L CIP induced by sublytic complement. Both ionophore- and complement-induced protection were blocked by treatment with protein kinase C (PKC) inhibitors. Calphostin C, sphingosine and GF109203X abrogated complement-induced protection almost completely, whereas Go6976 inhibited it only partially. Since Go6976 is a selective inhibitor of the conventional PKC type, it is proposed that sublytic complement doses activate both conventional and non-conventional PKC types. Immunofluorescence analysis of K562 cells demonstrated sublytic complement induced translocation of the conventional PKCalpha and PKCbetaII from the cytoplasm to the plasma membrane. These results indicate that PKC activation is an early obligatory signal in cell desensitization by sublytic C5b-9 or calcium ionophore. PMID- 10820373 TI - Thymic selection is influenced by subtle structural variation involving the p4 residue of an MHC class I-bound peptide. AB - The T lineage repertoire is shaped by opposing processes of positive and negative selection. To probe the specificity of selection, N15 TCR-transgenic (tg) recombinase-activating gene (RAG)-2(- / -) H-2(b) mice recognizing the VSV8 octapeptide RGYVYQGL bound to K(b) were utilized in conjunction with VSV8 variants differing only at the central p4 position. The V4I mutant octamer, like VSV8, induces negative selection of immature double-positive thymocytes on the beta(2)-microglobulin (beta(2)M)(+ / +) background and is a strong agonist for mature N15 T cells. In contrast, V4L or V4norvaline octamers promote positive selection in N15tg RAG-2(-/-) beta(2)M(-/-) H-2(b) fetal thymic organ culture and are weak agonists for N15 T cells. Hence, the absence of a p4 side chain Cbeta methyl group results in positive selection of the N15 TCR. Hydrophobicity of the p4 residues also modulates thymocyte fate: the positively selecting norvaline and leucine variants have one and two Cgamma-methyl groups, respectively, while the weakly selecting gamma-methylleucine p4 contains three Cgamma-methyl groups. Moreover, the most hydrophobic octamer containing p4 cyclohexylglycine substitution fails to select. Thus, for N15 and presumably other MHC class I restricted TCR, there is a high degree of structural specificity to peptide dependent thymic selection processes. PMID- 10820374 TI - IL-5 and eosinophils mediate the rejection of fully histoincompatible vascularized cardiac allografts: regulatory role of alloreactive CD8(+) T lymphocytes and IFN-gamma. AB - CD8(+) T lymphocytes are known to inhibit the development of eosinophilia and IL 5 synthesis in models of experimental lung disease. In transplantation, the rejection of fully mismatched cardiac allografts by recipients depleted of CD8(+) T cells is characterized by the recruitment of eosinophils in the rejected organs. We show here that this intragraft eosinophilia is dependent on the production of IL-5 since hearts transplanted into IL-5-deficient recipients depleted of CD8(+) cells did not contain eosinophils. More importantly, allograft survival was significantly extended in these animals. In mixed lymphocyte cultures (MLC), the presence of CD8(+) T cells in the responding cell population inhibited the secretion of IL-5. This inhibition was IFN-gamma dependent since adding neutralizing anti-IFN-gamma antibodies induced the production of IL-5. Furthermore, spleen cells isolated from IFN-gamma receptor (IFN-gammaR)-deficient mice secreted IL-5 upon allogeneic stimulation in primary MLC. In vivo, eosinophilia was observed in allografts rejected by IFN-gammaR-deficient recipients. On the contrary, grafts rejected by IFN-gammaR-deficient mice treated with neutralizing anti-IL-5 antibodies did not exhibit eosinophilic infiltration. Our study reveals the capacity of IL-5-secreting CD4(+) T cells and eosinophils to promote the rejection of heart allograft and demonstrates the importance of CD8(+) T cells and IFN-gamma in regulating this pathway of rejection. PMID- 10820375 TI - Tumor size at the time of adoptive transfer determines whether tumor rejection occurs. AB - Here we investigate the minimal requirements for induction of an anti-tumor response in CD8 T cells in vivo. We compare the efficacy of adoptive transfer of CD8 T cells with a transgenic TCR specific for the main cytotoxic T lymphocyte epitope of the influenza virus nucleoprotein (NP) on the growth of NP-expressing EL4 tumors under different conditions. In a setting in which tumor rejection is solely dependent on tumor-specific CD8 T cells, small immunogenic tumors fail to induce a rejection response, despite the fact that they are not ignored: tumor specific CD8 T cells are activated, differentiate into effector cells and infiltrate the tumor bed. Nevertheless, tumor rejection does not occur. In sharp contrast, the same immunogenic tumor, when growing as a large tumor mass, is rejected by transferred tumor-specific CD8 T cells. The main features which distinguish the rejection response to a large tumor mass from the response to a small tumor is that, in the latter case, activated CD8 T cells appear much later, and in much smaller numbers. Efficacy of adoptive transfer is thus dictated by the size of the tumor mass at the time of transfer. These findings predict that treatment of minimal residual disease with adoptive transfer will fail, unless vaccination is also provided at the time of transfer. PMID- 10820376 TI - Survival and cytokine polarization of naive CD4(+) T cells in vitro is largely dependent on exogenous cytokines. AB - Naive CD4(+) T cells differ from memory cells by their heightened expression of the disialoceramide recognized by antibody 3G11. 3G11(bright) cells respond well to immobilized anti-CD3 / anti-CD28 and to their cognate antigens but produce little or no IFN-gamma or IL-4 "acutely" and undergo cell death even in the presence of IL-2. They can be rescued by IL-4, IL-6 or IL-12. IL-6 is particularly notable since it is neutral in regard to Th1 / Th2 priming, allowing an assessment of the role of endogenous IL-4 in priming for IL-4 production. Naive TCR-transgenic BALB / c scid T cells cultured with an ovalbumin peptide and IL-4(- / -) antigen-presenting cells in the presence of IL-6 showed a modest degree of priming for IL-4 production if both IFN-gamma and IL-12 were neutralized. This priming is far less than that observed if IL-4 is added to the priming culture. These results indicate that IL-4 production as a result of TCR engagement is sufficient for only a minor component of the polarization observed when unseparated BALB / c CD4 T cell populations are primed or when IL-4 is intentionally added to the priming culture. PMID- 10820377 TI - Cytoskeletal protein tyrosine phosphatase PTPH1 reduces T cell antigen receptor signaling. AB - The subgroup of protein tyrosine phosphatases that contain an N-terminal ezrin-, radixin- and moesin homology (ERM) domain and a C-terminal catalytic domain is represented by three enzymes in Jurkat T cells, PTPH1, PTP-MEG1 and PTP36. These enzymes are located at the cytoplasmic face of the plasma membrane and may be involved in regulation of the membrane cytoskeleton, signal transduction, or both. Here we report that expression of PTPH1 in Jurkat T cells reduced the TCR induced activation of reporter genes encompassing parts of the IL-2 gene promoter and driven by nuclear factor of activated T cells plus activator protein-1. PTP MEG1 had a weaker inhibitory effect, while PTP36 had none. The catalytically inactive mutants PTPH1-CS and PTP-MEG1-CS lacked effects on gene transcription. Expression of active PTPH1 also reduced receptor-induced activation of Erk2 MAP kinase, its upstream activator, Mek, and the Jnk kinases. The effect of PTPH1 was reduced by deletion of its N-terminal ERM domain. We suggest that PTPH1 inhibits T cell activation by dephosphorylating membrane-associated targets involved in TCR signaling. PMID- 10820378 TI - BLNK is associated with the CD72/SHP-1/Grb2 complex in the WEHI231 cell line after membrane IgM cross-linking. AB - Tyrosine phosphorylation of CD72 strongly correlates with B cell antigen receptor signals leading to apoptosis. We have previously shown that CD72 carrying two immunoreceptor tyrosine-based inhibition motifs (ITIM) is an in vivo substrate of SHP-1. CD72 forms a complex with SHP-1 and Grb2 via its tyrosine-phosphorylated ITIM when the WEHI231 cell line, which is representative of immature B cells, undergoes apoptosis. The CD72 complex formation was also demonstrated in normal primary B cells, suggesting that the complex formation in apoptotic B cells is a universal mechanism. In this study, we further investigated the molecular components of the CD72 complex in WEHI231 cells in order to understand the molecular mechanism involved in the signaling pathway mediated through the complex. Our experiments demonstrate that BLNK, a recently identified adaptor molecule predominantly expressed in B cells, is associated with the CD72 complex via the Src homology 3 domain(s) of Grb2 in the cell line after membrane IgM (mIgM) engagement. The results suggest that the mIgM-mediated signal strongly correlates with the formation of the CD72 / SHP-1 / Grb2 / BLNK complex. PMID- 10820379 TI - Evidence of a role for galectin-1 in acute inflammation. AB - Galectin-1 (Gal-1), a member of a family of beta-galactoside-binding proteins, has been suggested to play key roles in immunological and inflammatory processes. The present study deals with the concept of an in vivo role for Gal-1 in acute inflammation by using the rat hind paw edema test. Local administration of Gal-1 (0.5, 2, 4 and 8 microg/ml) inhibited acute inflammation induced by bee venom phospholipase A(2) (PLA(2)) when it was injected 30 min before the enzyme or co injected together with PLA(2). The anti-inflammatory effect was prevented by a specific antibody, but independent of its carbohydrate-binding properties. In contrast, Gal-1 failed to inhibit histamine-induced edema. Histopathological studies showed a clear reduction of the inflammatory process when Gal-1 was injected before PLA(2), evidenced by a diminished number of infiltrated polymorphonuclear neutrophils and scarce degranulated mast cells. The anti inflammatory effect was also assessed in vitro, showing that Gal-1 treatment reduced prostaglandin E(2) secretion and arachidonic acid release from stimulated peritoneal macrophages. Results presented here provide the first evidence for a role of Gal-1 in acute inflammation and suggest that the anti-inflammatory effect involves the inhibition of both soluble and cellular mediators of the inflammatory response. PMID- 10820380 TI - IL-13 and TNF-alpha inhibit dual-tropic HIV-1 in primary macrophages by reduction of surface expression of CD4, chemokine receptors CCR5, CXCR4 and post-entry viral gene expression. AB - We show that IL-13 in the presence of TNF-alpha effected an equal or greater antiviral activity against a dual-tropic HIV-1 (R5X4) in macrophages. A temporary or continued exposure of macrophages to both cytokines significantly decreased the infection and replication of R5X4 HIV-1(89.6) (median, 128-fold, n = 9, p = 0.024) in macrophages as compared to untreated controls when analyzed over six decreasing multiplicities of infection. A quantitative flow cytometric assay revealed that IL-13 induced a significant (approximately 50 %) reduction in the number of CD4 and CC chemokine receptor 5 (CCR5) antibody binding sites while completely abrogating surface expression of CXC chemokine receptor 4 (CXCR4). In the presence of IL-13 and TNF-alpha, expression of CCR5 was completely abrogated while the expression of CD4 and CXCR4 remained significantly reduced as compared to untreated controls. A reduction in CD4 and HIV-1 coreceptors was associated with a decrease in reverse-transcribed viral DNA at 24 h post-infection. Quantification of viral gene expression using amphotropic MLV Env pseudotyped luciferase reporter viruses suggested that IL-13 inhibited HIV-1 gene expression within 24 h by up to 90 % in the presence or absence of TNF-alpha. In conclusion, our data suggest that IL-13 is a powerful counter-regulatory agent against TNF alpha-induced HIV-1 expression while also acting with TNF-alpha in inhibiting de novo infection of macrophages. PMID- 10820381 TI - Murine cytomegalovirus replication in salivary glands is controlled by both perforin and granzymes during acute infection. AB - The course of mouse cytomegalovirus (MCMV) infection was compared between wild type and mutant C57BL / 6 (B6) mice deficient in either RAG-2, perforin, granzyme A, granzyme B or combinations thereof at two time points post infection (p. i.). At day 15 p. i., virus titers were similarly elevated in salivary glands of all mutant, but not wild-type B6 mice and undetectable in lung and spleen tissues of any of the mouse strains. Significant pathological alterations were only seen in salivary glands and spleen from RAG2(- / -), but not in those from other mice whereas few inflammatory foci were observed in lung tissues of all mice except B6. At day 30 p. i., elevated virus titers were observed only in salivary glands, lung and spleen from RAG2(- / -), but in none of the other mice, and were accompanied by extended pathological alterations in all three organs. The data extend previous reports on the critical role of NK / CD8(+) T cells in the early control of MCMV infection by showing that both perforin and granzymes A / B contribute to viral elimination in salivary glands; however, neither of the three molecules alone seem to be indispensable for the final control of infection. PMID- 10820382 TI - Direct quantitation of rapid elimination of viral antigen-positive lymphocytes by antiviral CD8(+) T cells in vivo. AB - Lysis of infected cells by CD8(+) T cells is an important mechanism for the control of virus infections, but remains difficult to quantify in vivo. Here, we study the elimination kinetics of viral antigen-positive lymphocytes by antiviral CD8(+) T cells using flow cytometry and mathematical analysis. In mice acutely infected with lymphocytic choriomeningitis virus, more than 99.99 % of target cells were eliminated each day, corresponding to a half-life of 1.4 h. Even in mice exposed to virus 300 days previously, and with no ex vivo killing activity, 84 % of the target cells were eliminated per day. Unexpectedly, the elimination kinetics of antigen-positive lymphocytes was not significantly impaired in mice deficient in either perforin-, CD95 ligand- or TNF-mediated cytotoxicity. For viruses with a particular tropism for lymphocytes, such as Epstein-Barr virus or HIV, our results illustrate how effectively CD8(+) T cell-mediated elimination of target cells can potentially contribute to virus control and immunosuppression. PMID- 10820383 TI - Cytokine regulation of IL-12 receptor beta2 expression: differential effects on human T and NK cells. AB - The biological activities of IL-12 are mediated through a specific, high-affinity receptor composed of IL-12 receptor(R)beta1 and IL-12Rbeta2 subunits that exist primarily on T and NK cells. Remarkably, the expression of IL-12Rbeta2 on CD4(+) T cells in mouse and humans appears to be differentially regulated by IFN-gamma and IFN-alpha, respectively. Using an antibody specific for the human IL-12Rbeta2 subunit, the effect of IFN-gamma, IFN-alpha, IL-12 and IL-2 on the regulation of IL-12R expression and IL-12 responsiveness of human T and NK cells was assessed. The presence of IFN-alpha or IFN-gamma in cultures enhanced IL-12Rbeta2 expression of CD4(+) and CD8(+) T cells. The enhancing effect of IFN-alpha and IFN-gamma was independent of endogenous IL-12. Furthermore, the clearest effects of IFN-alpha and IFN-gamma on IL-12Rbeta2 expression on T cells were seen by abrograting the inhibition induced by the presence of IL-4 in cultures. In contrast to T cells, IFN-alpha and IFN-gamma had little effect on regulating IL 12Rbeta2 expression on human NK cells. Taken together, these data show that there is differential regulation of IL-12Rbeta2 expression by IFN-alpha and IFN-gamma on human T and NK cells. PMID- 10820384 TI - Yersinia enterocolitica-mediated translocation of defined fusion proteins to the cytosol of mammalian cells results in peptide-specific MHC class I-restricted antigen presentation. AB - Yersinia enterocolitica delivers a set of effector proteins [Yersinia outer proteins (Yop)] into the cytosol of target cells to modulate host cell signal transduction pathways required for the extracellular survival of the bacterium. Secretion and subsequent translocation of Yop across the eukaryotic cell membrane are achieved via a type III secretion system. About 50 - 100 amino acids of the N terminus of Yop are required for chaperone-directed secretion and translocation. In this study, it is demonstrated by immunoblot analysis of Yersinia-infected cultured epithelial cells that one ot these proteins, YopE, can serve as a molecular carrier to deliver protein fragments of the heterologous p60 antigen of Listeria monocytogenes into the cytosol of target cells. T cell activation assays revealed that the observed type III-mediated antigen translocation led to a p60 peptide-specific MHC class I-restricted antigen presentation. Efficient translocation and antigen presentation were strictly dependent on the co localized expression of hybrid YopE-p60 proteins and the YopE-specific chaperone SycE. These results suggest that the Yersinia type III secretion system may serve as an attractive tool for antigen delivery in Yersinia-based live vaccines to induce cellular immune responses. PMID- 10820385 TI - T lymphocyte and monocyte interaction by CD40/CD40 ligand facilitates a lymphoproliferative response and killing of Cryptococcus neoformans in vitro. AB - This study explored the role of CD40 / CD40 ligand (CD40L) in the induction of a lymphoproliferative response and killing of Cryptococcus neoformans in vitro. In our experimental system, monocytes exposed to C. neoformans were used as antigen presenting cells (APC) and co-cultured with autologous T cells. The results showed that CD40 / CD40L strongly regulated the blastogenic response of T cells to C. neoformans. The fungus up-regulated CD40 expression on APC. An acapsular strain appeared to be a better inducer than an encapsulated strain. Time course experiments showed optimal regulation of CD40 expression at 48 h of incubation. Blocking the interaction of CD40 on APC with CD40L on T cells using mAb to CD40L resulted in a significant inhibition of IFN-gamma production. The anti cryptococcal activity of monocytes was greatly influenced by the CD40 / CD40L interaction, and a positive correlation was found between nitric oxide secretion and enhanced killing of C. neoformans. Finally, the CD40 / CD40L interaction was critical for induction of optimal secretion of pro-inflammatory cytokines such as TNF-alpha and IL-1beta. These results indicate an important role for CD40 / CD40L interaction in inducing activation of T cells. Such cell-to-cell contact promotes anti-cryptococcal activity as well as secretion of pro-inflammatory cytokines by monocytes. PMID- 10820386 TI - B7.1 and B7.2 co-stimulatory molecules regulate crescentic glomerulonephritis. AB - The contribution of B7.1 and B7.2 co-stimulation to Th1-directed, cell-mediated renal injury was studied in a murine model of crescentic glomerulonephritis (GN) initiated by a "planted" antigen. Mice treated with anti-B7.2 monoclonal antibody (mAb), starting prior to disease initiation, developed more severe renal injury with increased glomerular crescent formation (p = 0.031), glomerular accumulation of T cells (p = 0.014) and proteinuria (p = 0.022) compared to mice treated with control antibodies. Mice treated with anti-B7.1 mAb had reduced crescent formation (p = 0.019) compared to control treated mice, but reductions in glomerular CD4(+) T cell accumulation and proteinuria were not statistically significant. B7. 1 mAb treatment significantly reduced all parameters of renal injury (above) compared to anti-B7.2 mAb treatment. Neither treatment altered the circulating antibody titer or cutaneous delayed type hypersensitivity to the nephritogenic antigen. Antibody subclasses and antigen-stimulated ex vivo splenocyte IL-4, IL-10 and IFN-gamma production did not indicate effects on Th subset responses. Treatment with CTLA4-Fc or combined treatment with anti-B7.1 and B7. 2 antibodies did not significantly attenuate crescentic GN. These data indicate that B7.1 and B7.2 are important co-stimulatory molecules involved in crescentic GN, which have opposing effects on disease development without altering the T helper cell subset response to the nephritogenic antigen. PMID- 10820387 TI - Selective involvement of the Fas (CD95)/Fas ligand pathway in bone marrow B cell progenitors. AB - B lymphocyte generation in bone marrow (BM) compensates for cell loses. The Fas / Fas ligand (FasL) pathway has been implicated in apoptosis of various cell types. Abnormalities of the Fas receptor or of FasL expression are associated with excessive T cell proliferation and autoimmunity. To examine the role of the Fas / FasL system in B cell differentiation, we created double-chimeric mice by transferring both C57BL / 6 (B6)-Fas(+) and lpr-FasL(+) BM cells into RAG-2(- / ) hosts. Equal numbers of stem cells were co-injected into sublethally irradiated recipients, and their progeny were studied by using antibodies directed against the B6-Ly5. 1(+)5.2(+) and lpr-Ly5.1(-)5.2(+) populations. A longitudinal study lasting for up to 6 months revealed that cells of the lpr phenotype dominated the B6 phenotype in the BM, as a result of their active proliferation. Analysis of the B cell compartment showed more lpr than B6 cells among immature HSA(hi)B220(lo) populations. In contrast, the lpr and B6 phenotypes were equally represented among mature B cells. BM transfer to second hosts indicated that B6 derived B cell progenitors were absent from the first host. These data suggest that activation of the Fas / FasL pathway disturbs the early steps of B cell development and might therefore contribute to the onset of autoimmune disorders. PMID- 10820388 TI - Induction of experimental autoimmune encephalomyelitis in C57BL/6 mice deficient in either the chemokine macrophage inflammatory protein-1alpha or its CCR5 receptor. AB - Macrophage inflammatory protein (MIP)-1alpha is a chemokine that is associated with Th1 cytokine responses. Expression and antibody blocking studies have implicated MIP-1alpha in multiple sclerosis (MS) and in experimental autoimmune encephalomyelitis (EAE). We examined the role of MIP-1alpha and its CCR5 receptor in the induction of EAE by immunizing C57BL / 6 mice deficient in either MIP 1alpha or CCR5 with myelin oligodendrocyte glycoprotein (MOG). We found that MIP 1alpha-deficient mice were fully susceptible to MOG-induced EAE. These knockout animals were indistinguishable from wild-type mice in Th1 cytokine gene expression, the kinetics and severity of disease, and infiltration of the central nervous system by lymphocytes, macrophages and granulocytes. RNase protection assays showed comparable accumulation of mRNA for the chemokines interferon inducible protein-10, RANTES, macrophage chemoattractant protein-1, MIP-1beta, MIP-2, lymphotactin and T cell activation gene-3 during the course of the disease. CCR5-deficient mice were also susceptible to disease induction by MOG. The dispensability of MIP-1alpha and CCR5 for MOG-induced EAE in C57BL / 6 mice supports the idea that differential chemokine expression patterns represent differences in disease mechanism that underlie various models of EAE, and possibly distinct patterns of pathology seen in MS. PMID- 10820389 TI - Stat6 activation and Th2 cell differentiation [correction of proliferation] driven by CD28 [correction of CD28 signals]. AB - CD28 engagement by specific monoclonal antibody (mAb) or binding of the natural ligands, CD80 and CD86, induces tyrosine phosphorylation of CD28, which in turn recruits and activates the signal transducer and activator of transcription 6 (Stat6). The Stat6 association with CD28 is specifically induced by CD80 or CD86 ligand binding and is not dependent upon the secretion of IL-4 or IL-13. Activated Stat6 translocates to the nucleus and binds to a Stat6-responsive element on the human IL-4 promoter. CD28 ligation induces Stat6-dependent transcriptional activation of a reporter gene under the control of a multimerized Stat6-responsive element fused to an essential part of the IL-4 promoter. Primary stimulation of naive CD4(+) T cells with anti-CD28 mAb in the presence of IL-2, but in the absence of anti-CD3 mAb induces preferential production of IL-4 and expression of CCR4 mRNA after secondary stimulation with anti-CD3, indicating the preferential differentiation of Th2 cells. These findings suggest that initial IL 4 production required for commitment of naive T cells toward Th2 cells may be provided in response to signals delivered via CD28 by antigen-presenting cells. PMID- 10820390 TI - IL-12 selectively regulates STAT4 via phosphatidylinositol 3-kinase and Ras independent signal transduction pathways. AB - IL-12 is an important immunomodulatory cytokine that induces tyrosine phosphorylation and activation of the signal transducer and activator of transcription (STAT)4. IL-12 induces sustained activation and nuclear translocation of STAT4 and this regulatory process is coupled to both tyrosine and serine phosphorylation of this molecule. IL-12-activated tyrosine kinases are the Janus kinases Jak2 and Tyk2 but little is known about IL-12 regulation of serine kinases. The object of the present study was to explore the role of mitogen-activated protein kinases (MAPK) Erk1 and Erk2 and phosphatidylinositol 3 kinase (PI3K) in STAT4 regulation. Here we show that the IL-12-induced STAT4 serine kinase is not sensitive to inhibitors of the PI3K or MAPK Erk1,2. Moreover, IL-12 activation of STAT4 in human peripheral blood-derived T cells is not accompanied by stimulation of the Ras guanine nucleotide binding cycle or stimulation of MAPK Erk1,2 or initiation of the PI3K signaling pathways. IL-12 is unable to initiate the serine phosphorylation of STAT 1 and 3. This reveals that the STAT1, 3 and 4 serine kinases are not coordinately regulated in human T cells and that IL-12 must regulate serine phosphorylation of STAT4 by a kinase network distinct to the MAPK Erk1,2 or PI3K pathways. PMID- 10820391 TI - Immunosuppressant FTY720 inhibits thymocyte emigration. AB - One major role of the thymus is to provide the peripheral immune system with mature T cells, but the mechanisms involving the cellular export are not fully understood. In this study, we examined the ability of a novel immunosuppressive reagent, FTY720, to inhibit T cell export from the thymus. Daily administration of FTY720 at a dose of 1 mg / kg resulted in a marked decrease in the number of peripheral blood T lymphocytes. In the thymus, long-term daily administration of FTY720 caused a three- to fourfold increase in the proportion of mature medullary thymocytes (CD4(+)CD8(-) and CD4(-)CD8(+)) as well as a slight decrease in the double-positive cell (CD4(+)CD8(+)) ratio. Phenotypic analysis (TCRalpha beta, H 2K(d), CD44, CD69 and CD24) revealed that these increased subsets represent possible peripheral recent thymic emigrants. High level expression of L-selectin by these subsets further suggests that they were prevented from leaving the thymus. By intrathymic labeling with fluorescein isothiocyanate, only one fourth of labeled cells could be detected in the lymph nodes and in the spleen of FTY720 treated mice compared to saline-treated control mice. Taken together, these results suggest that the immunosuppressive action of FTY720, at least in part, could be due to its inhibitory effect on T cell emigration from the thymus to the periphery. PMID- 10820392 TI - IL-12 enhances IL-2 function by inducing CD25 expression through a p38 mitogen activated protein kinase pathway. AB - We previously showed that an IL-2 mutant, Q126D, could induce T cells to proliferate to the same extent as wild-type IL-2 but was unable to sensitize T cells to activation-induced cell death (AICD). Here we show that the partial signaling of Q126D is attributable to its inability to up-regulate the IL-2 receptor alpha chain (CD25). IL-12, which can up-regulate CD25 expression, enhances the ability of Q126D to induce AICD sensitivity and proliferation. IL-12 synergism with Q126D is dependent on CD25 up-regulation because the synergism is absent in CD25-deficient T cells. Inhibition of IL-12-induced up-regulation of CD25 by a p38 mitogen-activated protein (MAP) kinase inhibitor, SB203580, also ablates the synergism between IL-12 and Q126D. Although CD25 is important for IL 2-induced proliferation and AICD sensitivity, it is not absolutely required because a high concentration of IL-2 can overcome the requirement for CD25. Under physiological concentrations of IL-2, CD25 expression is critical for the function of IL-2. IL-12 can enhance the function of IL-2 by up-regulating CD25 in a p38 MAP kinase-dependent manner. PMID- 10820393 TI - Expression of functional IL-2 receptors on mature splenic dendritic cells. AB - We report here the expression of functional IL-2 receptor (IL-2R) on mature splenic dendritic cells (DC) and synergistic effect of IL-2 on IFN-gamma production by DC. IL-2 augmented IL-12-dependent IFN-gamma production by DC purified from both splenocytes of wild-type and anti-asialoGM1 Ab-treated Rag-2( /-) splenocytes devoid of T, B, NK and NKT cells. A neutralizing mAb against IL 2Ralpha blocked such enhancing effect of IL-2 on IFN-gamma production, indicating the presence of functional IL-2R on DC. Synergistic effects of IL-2 were also observed on IFN-gamma production by DC stimulated through CD40 or MHC class II, suggesting that T cell-derived IL-2 can act on DC during antigen presentation. Furthermore, we provide evidence that DC produce IFN-gamma during interaction with allogeneic CD4(+) T cells from IFN-gamma(-/-) mice. These results suggest that IL-2 produced by naive T cells upon antigen stimulation is an important factor during Th0 to Th1 differentiation by inducing IFN-gamma from DC. PMID- 10820394 TI - Tracing and characterization of the low-avidity self-specific T cell repertoire. AB - It is well established that expression of self antigens results in the deletion of the functional high-avidity self-specific T cell repertoire. Due to the low frequency of naturally occurring low-avidity self-specific T cells, a detailed evaluation of their ability to survive and differentiate into effector and memory populations in vivo has yet to be obtained. We here employ tetramer technology to characterize and determine the in vivo fate of a self-specific CD8(+) T cell population specific for a ubiquitously expressed T cell epitope. We find that in influenza nucleoprotein (NP)-transgenic mice (B10NP mice) an oligoclonal population of NP(366 - 374)-specific T cells can be triggered by live influenza virus exposure. The main hallmark of this self-specific T cell population is its diminished avidity for the tetrameric MHC / NP peptide complex. These low-avidity T cells are not deleted and do not down-regulate their antigen or CD8 receptors, and exhibit cytolytic activity towards tumor cells expressing NP endogenously. Strikingly, a secondary influenza infection generates a typical memory response in the low-avidity repertoire. The observation that low-avidity T cells persist in vivo and can differentiate into memory T cells underscores their potential role in anti-tumor immunity. PMID- 10820395 TI - Variability of invariant mouse CD3epsilon chains detected by anti-CD3 antibodies. AB - Current models of the TCR / CD3 complex assume that, in mature peripheral T lymphocytes, variability is restricted to the alpha beta (or gamma delta) chains of the TCR heterodimer responsible for antigen recognition, whereas the CD3 polypeptides involved in signal transmission are invariant. Here we show that mouse CD4(+) T lymphocytes and T cell lines are bound with different avidity by anti-CD3 monoclonal antibodies. These findings cannot be accounted for by allelic differences between CD3 chains, by the nature of the TCR chains, or by the ratio of CD3epsilon delta to CD3epsilon gamma chain pairing. Rather, they are linked to heterogeneity of the N-terminal region of CD3epsilon chains, as detected by peptide-specific antibodies. In turn, these differences among CD3epsilon chains correlate with variations in the strength of TCR / CD3 interaction. N-terminal CD3epsilon heterogeneity is not due to alternative splicing mechanisms, but rather involves digestion by metalloproteases, as suggested by reverse transcription-PCR amplification and by the effect of protease inhibitors, respectively. Based on these data, we propose a model linking CD3epsilon N terminal variability with altered CD3 recognition by monoclonal antibodies and TCR / CD3 interaction. This model suggests the possibility of distinct spatial arrangements of the TCR / CD3 complex. PMID- 10820396 TI - Definition of polymorphic residues on killer Ig-like receptor proteins which contribute to the HLA-C binding site. AB - Killer cell immunoglobulin-like receptors (KIR) bind HLA class I proteins in an allele- and locus-specific manner. This report describes the use of transfectants expressing recombinant chimeric proteins, comprising the extracellular portions of KIR molecules and the transmembrane and cytoplasmic tails of CD3-zeta, to create an in vitro system in which signaling is readily measured and that preserves the specificity of the KIR / HLA-C interaction. The identity of the amino acid residues on the KIR molecule important for binding to the HLA protein is not well understood; although some KIR2D residues involved in HLA-C recognition have been identified, their relative importance and whether other amino acids contribute to binding was unclear. This novel system was used to study, by site-directed mutagenesis, the role of various amino acids in KIR binding to HLA-C ligand. The data presented here show that while multiple polymorphic residues contribute to the HLA-C binding site on KIR proteins, two clusters of polymorphic residues define the group allotype specificity of HLA-C binding to a KIR2D molecule. PMID- 10820397 TI - Hapten-induced colitis associated with maintained Th1 and inflammatory responses in IFN-gamma receptor-deficient mice. AB - IFN-gamma is a potent pro-inflammatory cytokine thought to be involved in the pathogenesis of Crohn's disease. To further define the role of IFN-gamma in intestinal inflammation, we studied the effects of intra-colonic 2,4,6 trinitrobenzene sulfonic acid (TNBS) instillation in mice with a functionally inactivated IFN-gamma receptor 1 (IFN-gammaR1(- / -)). Our results indicate that IFN-gamma is not necessary for the induction of hapten-induced colitis: after TNBS administration both wild-type and IFN-gammaR1(- / -) mice lost body weight, and the histological features of TNBS-induced colitis were comparable. Colons of IFN-gammaR1(- / -) mice contained a greater number of cells, represented by macrophages and CD4(+) T cells; caudal lymph node cells produced more IFN-gamma and TNF-alpha upon stimulation in vitro. Moreover, IL-18 and IL-12 p40 RNA levels were comparably up-regulated after TNBS treatment in IFN-gammaR1(- / -) wild-type mice. These findings demonstrate that IFN-gamma is dispensable for the development of TNBS-induced colitis. Importantly, the production of Th1 cytokines (e. g. IFN-gamma and TNF-alpha) by caudal lymph node T lymphocytes was enhanced rather than decreased in IFNgammaR1(- / -) mice with no evidence for default Th2 development. PMID- 10820398 TI - Proteolytic processing of peptides in the lumen of the endoplasmic reticulum for antigen presentation by major histocompatibility class I. AB - We have tested the hypothesis that MHC class I molecules are actively involved as protease in the production of natural MHC class I ligands. First, the structure of a class I molecule was analyzed for homology with catalytic sites of known proteases. While several clusters of amino acids in the restriction element resembled protease active sites, structural discrepancies and the influence of nearby residues suggest that these sites are unlikely to have protease activity. Second, we have tested the presentation of viral cytotoxic T cell determinants with affinity for the same restriction element (H-2K(d) or K(k)), when targeted as tandem peptides into the endoplasmic reticulum. Peptide transporter-defective cells were used to exclude cleavage of the tandem peptides by cytosolic proteases. Cleavage by signal peptidase of the tandem peptides was ascertained. The C-terminal peptides in the tandem arrays were almost exclusively presented, suggesting that an aminopeptidase in the endoplasmic reticulum degraded the N terminally positioned peptides. This result is inconsistent with an MHC class I catalyzed cleavage following binding of longer peptides in the cleft of the restriction elements. Finally, we conclusively show that an aminopeptidase in the endoplasmic reticulum is also involved in antigen presentation in cells with a functional peptide transporter. PMID- 10820399 TI - Mouse complement components C4 and Slp act synergistically in a homologous hemolytic C4 assay. AB - Goals of the present study were to compare the hemolytic activities of mouse C4 and Slp in a homologous system and to study a possible interaction between these proteins during complement activation. As reagents for mouse C4 and Slp, we used serum of C4(- / -) knockout C57BL / 6 (C4(-) / Slp(-)) mice and sensitized rabbit erythrocytes as target cells. Sera to be tested contained none, either of the two or both proteins. We found that C4(-) / Slp(+) serum has some hemolytic C4 activity, but less than C4(+) / Slp(-) serum. Comparing C4 activities of C4(+) / Slp(-) and C4(+) / Slp(+) sera, we found a threefold enhanced activity in double positive serum. Hemolytic C4 levels of mixtures of solely C4- and Slp-sufficient sera did not overlap with expected C4 levels, but rather these sera showed synergy. This explains the enhanced activity of double-positive serum. Similar results were observed for total complement activation. In conclusion, Slp has measurable, but poor C4 activity as compared with mouse C4. Using our homologous system, we showed that the enhanced classical pathway activity of double-positive sera is most probably based on synergy between C4 and Slp. Our results answer an old question as to why C4(+) / Slp(+) mice have higher complement levels than C4(+) / Slp(-) mice. PMID- 10820400 TI - Vgamma9/Vdelta2 T lymphocytes reduce the viability of intracellular Mycobacterium tuberculosis. AB - An effective immune response against the intracellular pathogen Mycobacterium tuberculosis is strictly dependent on T cell activation. Although this protective response mainly depends on local release of pro-inflammatory cytokines by Th1 CD4(+) T cells, contribution of Vgamma9 / Vdelta2 T lymphocytes to immune protection against this pathogen is suggested by the antimycobacterial reactivity of this subset and its ability to produce large amounts of Th1 cytokines. Here we show that Vgamma9 / Vdelta2 T lymphocytes kill macrophages harboring live M. tuberculosis. The cytotoxic activity of Vgamma9 / Vdelta2 T lymphocytes was not MHC class I or class II restricted but was blocked by anti-TCR monoclonal antibodies, thus indicating that it involved specific interaction between the TCR and the target cell. The cytotoxicity of Vgamma9 / Vdelta2 T lymphocytes was not mediated by TNF-alpha or Fas-Fas ligand, but was shown to occur through a granule dependent mechanism that resulted in reduction of the viability of intracellular bacilli. Perforin was shown to play an important role in killing of both infected macrophages and intracellular mycobacteria. These data strongly suggest that Vgamma9 / Vdelta2 T lymphocytes contribute to the host defense against M. tuberculosis infection. PMID- 10820401 TI - Increased frequency of RAG-expressing, CD4(+)CD3(low) peripheral T lymphocytes in patients with defective responses to DNA damage. AB - Accumulating evidence indicates that peripheral lymphocyte variants with altered antigen receptor expression may be capable of expressing recombination-activating genes (RAG). We and others recently observed functional RAG gene products in mature T cells with defective TCR expression (MacMahan and Fink, Immunity 1998. 9: 637 - 647; Lantelme et al., J. Immunol., 2000. 164: 3455 - 3459). Here, the association between TCR expression and RAG activity was assessed further in lymphocytes from patients with defective responses to DNA damage. We show that T cells with altered TCR surface expression are present in increased numbers in these patients and that they express RAG genes. The finding of RAG gene expression by TCR variants suggests the possibility that secondary V(D)J rearrangements could be induced in these cells to rescue their defective phenotype and cellular function. Moreover, as V(D)J recombination has been implicated in chromosome translocations involving antigen receptor genes, we discuss a possible relationship between altered TCR expression, RAG activity and the frequent lymphoma-specific translocations observed in these patients. PMID- 10820402 TI - Non-concordance of CVS and liver glycine cleavage enzyme in three families with non-ketotic hyperglycinaemia (NKH) leading to false negative prenatal diagnoses. AB - We report three false negative prenatal diagnostic results, using direct measurement of glycine cleavage enzyme activity in uncultured chorionic villus tissue from 290 pregnancies at risk for non-ketotic hyperglycinaemia (NKH). Testing was done by two centres: Vancouver, Canada and Lyon, France. One false negative result had activity near the lower limit of the normal range but two samples gave completely normal results well within the control range. All three pregnancies continued and the three children were born affected with NKH. Because of the first result, we now counsel that there is a grey zone of uninterpretable activity where affected and normal enzyme values overlap. Because of the other two results we now counsel that there is an approximately 1% chance of a pregnancy with a normal CVS activity resulting in an affected child. The clinical and biochemical findings in the three families are discussed. PMID- 10820403 TI - DNA based prenatal testing for the skin blistering disorder epidermolysis bullosa simplex. AB - Epidermolysis bullosa simplex (EBS) is a skin fragility disorder in which mild physical trauma leads to blistering. The phenotype of the disorder is variable, from relatively mild affecting only the hands and/or feet, to very severe with widespread blistering. For the severest forms of EBS there is a demand for prenatal diagnosis which until now has involved a fetal skin biopsy in the second trimester. The identification of mutations in the genes encoding keratins K5 and K14 as the cause of EBS opens up the possibility of much earlier diagnosis of the disease. We report here four cases in which prenatal testing was performed. In three of the cases the genetic lesions were unknown at the start of the pregnancy, requiring the identification of the causative mutation prior to testing fetal DNA. In two of the four cases novel mutations were identified in K14 and in the two remaining families, a previously identified type of mutation was found. Fetal DNA, obtained by chorionic villus sampling or amniocentesis, was analysed for the identified mutations. Three of the DNA samples were found to be normal; a mutant K14 allele was identified in the fourth case and the pregnancy was terminated. These results demonstrate the feasibility of DNA-based prenatal testing for EBS in families where causative mutations can be found. PMID- 10820404 TI - Prenatal diagnosis of beta-thalassaemia in Pakistan: experience in a Muslim country. AB - A service for prenatal diagnosis of beta-thalassaemia was introduced in Pakistan in May 1994. Two renowned Islamic scholars, consulted before the service was introduced, ruled that a pregnancy can be terminated if the fetus is affected by a serious genetic disorder, and if termination is before 120 days (17 weeks) of gestation. During the first 3(1/2) years of the service 300 couples requested the test. Almost all the couples had been informed by their treating doctors. Most diagnoses were made between 10 and 16 weeks of gestation, and only 15 (5%) were reached after the 16th week. DNA analysis was by the amplification refractory mutation system (ARMS). A multiplex ARMS was developed in which three primer combinations identified the mutations in 91.5% of the couples. In 13 couples (4. 3%) linkage analysis was required for the fetal diagnosis. In 47/53 (88.7%) women carrying an affected fetus the pregnancy was terminated. In six cases it was declined principally on religious grounds. Postnatal confirmation of the prenatal diagnosis was possible in 117 unaffected children. One year after the start of the service, interviews with 141 couples with an affected child showed that 72% knew of the availability of prenatal diagnosis. Thirty-two of the informed couples had had a pregnancy, but only 18 (56%) used prenatal diagnosis. The main reasons for non-utilization of prenatal diagnosis were the cost of the test and fear of undergoing the test, though some gave no clear explanation. This study demonstrates that prenatal diagnosis is feasible and acceptable in a Muslim country such as Pakistan. PMID- 10820405 TI - Prenatal diagnosis of inherited satellited non-acrocentric chromosomes. AB - We report on the prenatal diagnosis of two sib female fetuses with a satellited short arm of chromosome 4 and a male fetus with a satellited long arm of chromosome X. The first two fetuses had a cryptic balanced translocation (4;15)(p16;p11.1) inherited from a mother carrying a satellited 4p and having an affected child with the Wolf-Hirschhorn syndrome. The third fetus had a satellited Xq, with a deletion of subtelomeric region of Xq. The mother was subsequently found to have the same satellited Xq but without the presence of a reciprocal translocation. She decided to continue the pregnancy. The proband with a satellited Xq manifested developmental delay, mental retardation, hypertelorism, ptosis of one eye, low-set ears, and hearing disturbance at age 6 months. Fluorescence in situ hybridization (FISH) with a specific telomeric or subtelomeric probe, and genetic marker analyses were used to confirm the diagnosis. Pregnant women with satellited non-acrocentric chromosomes are at risk for carrying fetuses with chromosome abnormalities. If the X chromosome is involved, the fetuses can be affected with X-linked recessive disorders including mental retardation. Detailed genetic counselling, cytogenetic studies, FISH and genetic marker analyses are useful in prenatal detection of abnormal chromosome rearrangements. PMID- 10820406 TI - Maternal serum free beta-hCG and PAPP-A in fetal sex chromosome defects in the first trimester. AB - We have studied maternal serum free beta-hCG and PAPP-A, and fetal nuchal translucency (NT) in a series of 46 cases of fetal Turner's syndrome, 13 cases of other sex chromosomal anomalies and compared these with 947 control pregnancies in the first trimester. In cases of Turner's syndrome (45,X) the median fetal NT was significantly higher than in controls (4.76 MoM), the median PAPP-A was significantly lower (0.49 MoM), whilst the free beta-hCG was not significantly different (1.11 MoM). For NT, 93% (43/46) of cases were equal to or greater than the 95th centile of controls, for PAPP-A 35% (16/46) of cases were less than or equal to the 5th centile of controls and for free beta-hCG 15% (7/46) of cases were equal to or greater than the 95th centile of controls. For other sex chromosomal anomalies (47XXX, XXY, XYY) the median NT was increased (2.07 MoM) whilst PAPP-A was not significantly decreased (0.88 MoM) and free beta-hCG was not significantly different (1.07 MoM) from controls. Using a previously derived multivariate risk algorithm for trisomy 21, incorporating NT, PAPP-A, free beta hCG and maternal age, 96% of the Turner's cases and 62% of the other sex chromosomal anomalies would have been identified. PMID- 10820408 TI - Prenatal diagnosis of alobar holoprosencephaly by two-dimensional and three dimensional ultrasound. AB - The aim of this study is to evaluate the sonographic characteristics of alobar holoprosencephaly (AH) in utero. Seventeen cases were diagnosed at 16-30 weeks' gestation by two-dimensional and three-dimensional ultrasound from October 1994 to December 1998. In this series, the prenatal prevalence was 1 out of 415 and the detection rate of AH by prenatal ultrasound was 100%. Eleven cases (64.7%) had concurrent facial anomalies. Cleft lip and hypotelorism were the most common associated facial anomalies (72.7%). Two cases (11.8%) were twin pregnancy with one fetus affected. We present one of the largest series in the literature and compare it with previous reports. From this series, we concluded that: (1) intracranial findings, including monoventricle, fused thalami, and the absence of midline structures, were reliable sonographic characteristics for prenatal diagnosis; (2) cleft lip and hypotelorism seemed to be more common associated facial malformations than cyclopia or cebocephaly with AH; and (3) although three dimensional ultrasound did not change the diagnosis of AH made by two-dimensional ultrasound in this series, three-dimensional ultrasound does assist in defining the severity and extent of AH. PMID- 10820407 TI - Screening for trisomies 21 and 18 with maternal serum placental isoferritin p43 component. AB - A component of placental isoferritin, p43, is an immuno-regulatory protein associated with suppression of the immune system. Maternal serum p43 levels increase throughout pregnancy and low serum levels have been associated with various pathological pregnancies, particularly those with a defect of placentation. We measured maternal serum p43 retrospectively in banked samples from 42 Down syndrome, 20 Edwards' syndrome and 281 unaffected pregnancies to assess its screening potential in both the first and second trimesters. The median maternal serum p43 level in Down syndrome was 1.58 times higher than that in the unaffected pregnancies (p=0.01, two-tail). The median level was slightly, but not significantly, reduced in Edwards' syndrome. Statistical modelling, including parameters for alpha-fetoprotein, free beta-human chorionic gonadotrophin, and unconjugated oestriol suggested that it might have a role in Down syndrome screening when combined with two or three of these markers. Larger scale studies are now needed. PMID- 10820409 TI - Diaphragmatic hernia as the first echographic sign in Apert syndrome. AB - We report left diaphragmatic hernia as the first presenting echographic sign in a malformed male fetus with Apert syndrome. Molecular studies confirmed the presence of a mutation in the FGFR2 gene (Ser 252 Trp). PMID- 10820410 TI - The use of transferrin for enrichment of fetal cells from maternal blood. AB - Iron loaded transferrin (holotransferrin) was used for enrichment of fetal cells from peripheral blood of pregnant women. Cord blood samples were used to evaluate enrichment efficacy of single and double MACS separations. Blood samples were obtained from 10 pregnant women prior to chorion villus sampling (CVS). Erythroblasts and other mononuclear cells were isolated by triple-density gradient centrifugation. Fetal cells were further enriched by positive magnetic sorting (VarioMACS) using biotinylated transferrin and streptavidin conjugated to magnetic microbeads. The isolated cells were analysed with dual-colour in situ hybridization (FISH) with X- and Y-chromosome specific probes. Male fetuses were correctly identified in three out of four (75%) pregnancies and female fetuses in six out of six (100%) pregnancies. By using transferrin instead of antibodies to the transferrin receptor to label and enrich fetal cells, we believe that unspecific binding attributed to immunological labelling can be minimized. The results indicate that transferrin may be an alternative to antibodies to transferrin receptor for separation of fetal cells from maternal blood. PMID- 10820411 TI - Screening for trisomy 13 by fetal nuchal translucency and maternal serum free beta-hCG and PAPP-A at 10-14 weeks of gestation. AB - In 42 cases of trisomy 13 at 10-14 weeks of gestation, compared with 947 controls, the median multiple of the median (MoM) of maternal serum free beta human chorionic gonadotrophin (beta-hCG) and pregnancy associated plasma protein A (PAPP-A) was significantly decreased (0.506 MoM and 0.248 MoM respectively), whilst fetal nuchal translucency was increased (2.872 MoM). In 38% and 71% of cases of trisomy 13 maternal serum free beta-hCG and PAPP-A was below the 5th centile of the appropriate normal range for gestation and in 62% of cases the nuchal translucency was above the 95th centile. When combined together in a multivariate algorithm with maternal age, 90% of cases of trisomy 13 could be detected at a 0.5% false positive rate or 84% at a 0.1% false positive rate. We conclude that specific trisomy 13 risks should be part of developing risk algorithms combining maternal serum biochemistry and nuchal translucency for use in first trimester screening alongside those for trisomy 21 and trisomy 18. PMID- 10820412 TI - Apparent confined placental mosaicism of trisomy 16 and multiple fetal anomalies: case report. AB - Trisomy 16 is frequently found confined to the placenta (confined placental mosaicism (CPM)), with a structurally normal fetus. In some cases of trisomy 16, the fetus has uniparental disomy for chromosome 16 (UPD16) which is associated with intrauterine growth restriction (IUGR) and fetal anomalies. We report a case of apparent confined placental mosaicism for trisomy 16, using standard cytogenetic techniques, but with multiple fetal abnormalities including congenital diaphragmatic hernia in which there was no evidence of UPD in the disomic tissues examined. Subsequent examination of fetal tissues using fluorescent in situ hybridization (FISH) demonstrated low levels of mosaicism for trisomy 16 in all the tissues examined. The use of FISH permits identification of mosaicism which conventional techniques may not identify. PMID- 10820413 TI - Prenatal ultrasound of regional akinesia with Pena-Shokier phenotype. AB - This report describes sonographic features of the Pena-Shokeir phenotype secondary to regional akinesia at 28 weeks of gestation with maternal perception of good fetal movement. The diagnosis was based on the findings of no fetal activity in some parts of the body (upper limbs, a part of the face, and thorax), with deformation sequence of fixed flexion at wrist, elbow, and shoulder joints, fixed open mouth, fetal growth restriction, lung hypoplasia, polyhydramnios and normal chromosome study. Surprisingly, fetal akinesia involved only the upper limbs, a part of the face, and chest, whereas the lower limbs were completely normal in both morphology and activity. With vibroacoustic stimulation, the response of fetal heart rate acceleration, gross body movement including fetal head, spine and lower limb movement were observed, but the movement of the upper limbs and fetal breathing were completely absent. Spontaneous labour and delivery occurred at 29 weeks' gestation. Postnatal autopsy confirmed the prenatal findings. In conclusion, prenatal sonography plays an important role not only in detecting Pena-Shokeir phenotype but also in providing the detailed pattern of fetal akinesia. Finally, this case reaffirms the concept that function is an integral part of normal development. PMID- 10820414 TI - Complex genetic counselling and prenatal analysis in a woman with external ophthalmoplegia and deleted mtDNA. AB - Single large mitochondrial DNA deletions (DeltamtDNA) are usually spontaneously occurring and cause a wide variety of symptoms, ranging from severe infantile multisystem disorders to adult onset progressive external ophthalmoplegia (PEO). There is always heteroplasmy with a mixture of normal and mutant mtDNA and the levels usually vary widely between tissues. There is at present insufficient scientific basis for accurate genetic counselling of women with DeltamtDNA, but it is reasonable to assume that DeltamtDNA can be transmitted if it is present in the female germ cells. Here, we present the results of prenatal analysis in a woman with DeltamtDNA and PEO. No DeltamtDNA was detected by Southern blot and PCR analyses of chorionic villi from the first trimester of pregnancy, in cord blood obtained at birth or in peripheral blood from the child at six months of age. This makes it unlikely that the child will develop a severe infantile mitochondrial disorder due to transmission of high levels of DeltamtDNA. However, the complex mitochondrial genetics and the limited access to human tissues makes it impossible to exclude transmission of low levels of DeltamtDNA that possibly could cause disease later in life. PMID- 10820415 TI - Survival of Down syndrome in utero. PMID- 10820416 TI - Fetal hepatic haemangioendothelioma: a new association with elevated maternal serum alpha-fetoprotein. PMID- 10820417 TI - Trisomy 8 in chorionic villi-unpredictable results in follow-up. PMID- 10820418 TI - Serum screening in a Japanese population. PMID- 10820419 TI - Current awareness in prenatal diagnosis. AB - In order to keep subscribers up-to-date with the latest developments in their field, John Wiley & Sons are providing a current awareness service in each issue of the journal. The bibliography contains newly published material in the field of prenatal diagnosis. Each bibliography is divided into 17 sections: 1 Books, Reviews & Symposia; 2 General Interest; 3 Normal Fetal Development; 4 Gametogenesis and Pre-implantation Diagnosis; 5 First Trimester Diagnosis; 6 Second Trimester Diagnosis; 7 Fetal Diagnosis by Ultrasound and Other Imaging; 8 Maternal Screening; 9 Screening for Carriers of Genetic Abnormality; 10 Technological Developments; 11 Confined Placental Mosaicism and Uniparental Disomy; 12 Molecular Cytogenetics; 13 Fetal Cells in Maternal Circulation; 14 Fetal Therapy; 15 Psychosocial Aspects; 16 Epidemiology and Environmental Factors; 17 Developmental Pathology. Within each section, articles are listed in alphabetical order with respect to author. If, in the preceding period, no publications are located relevant to any one of these headings, that section will be omitted. PMID- 10820420 TI - Anticarcinogenicity potential of spinasterol isolated from squash flowers. AB - Spinasterol, an antimutagen, was isolated from squash flowers by solvent partitioning and repeated vacuum liquid chromatography. Spinasterol was then tested for its anticarcinogenic potential by using the mouse skin tumor assay. There was a 90% skin tumor incidence for the positive control group (DMBA + croton oil + acetone). At a concentration of 15.0 microg/0.2 ml acetone, spinasterol decreased the incidence of skin tumors by 55.6% and decreased the number of tumors by 65.0% when applied immediately after croton oil. Hence, spinasterol showed antitumorigenic potential. It is not a co-carcinogen nor a co tumor promoter as there was no increase in the incidence of skin tumors after spinasterol application. Teratogenesis Carcinog. Mutagen. 20:99-105, 2000. PMID- 10820421 TI - Restriction site mutation (RSM) analysis of 2-acetylaminofluorene (2-AAF)-induced mouse liver mutations and comparison with the measurement of in vivo micronucleus induction in the bone marrows of (2-AAF)-treated mice. AB - We report here the successful application of the restriction site mutation (RSM) assay in detecting 2-acetylaminofluorene (2-AAF)-induced mouse liver mutations. A total of seven 2-AAF-induced liver mutations were detected out of a total of 304 analyses performed on 2-AAF-treated liver tissue. No mutations were detected in the 190 RSM analyses performed on untreated liver tissue. The 2-AAF-induced point mutations comprised 60% GC-->TA transversions, 30% GC-->AT transitions, 10% GC- >CG transversions, and 1 insertional event was also detected. All seven mutations were detected in intron 6 of the mouse p53 gene, with no mutations detectable in exons 4 or 5, supporting our previous data on the greater mutability of intron regions. In addition to the RSM analysis, we also report the application of the in vivo bone marrow micronucleus assay in detecting the clastogenicity of 2-AAF. We detected a small, but statistically significant, increase in the number of micronuclei induced by 2-AAF, but only after 2,000 cells were scored. This also confirms previous data showing that 2-AAF is a weak clastogen. Finally, we attempted to compare the sensitivity of the two assays to 2-AAF-induced genotoxicity, as had been previously undertaken with ENU. Both assays detected genotoxicity in their respective tissues; however, different endpoints were analysed. The RSM assay appears to be more adaptable than the micronucleus assay, due to its tissue and organism independence and has the potential to provide more molecular information on genotoxicity. Teratogenesis Carcinog. Mutagen. 20:107 117, 2000. PMID- 10820422 TI - DNA damage and repair in human lymphocytes exposed to three anticancer platinum drugs. AB - Cisplatin is a widely used anticancer drug, but its application is limited due to severe side effects. To reduce these effects, many other platinum drugs have been synthesized. In the present work comparative analysis of the toxicity of cisplatin, oxoplatin, and a conjugate (NH(3))(2)Pt(SeO(3)) (Se-Pt) in terms of cell viability, DNA binding, and DNA damage and repair in human lymphocytes was performed using the Trypan blue exclusion test, atomic absorption spectroscopy, and the comet assay, respectively. Cisplatin and oxoplatin did not cause a significant change in the viability of the lymphocytes even at the highest used concentration (750 microM), but the conjugate dramatically diminished viability at 100 microM only about 60% of the lymphocytes were viable (P < 0.05), and at 750 microM, less than 20% (P < 0.001). Se-Pt bound to isolated DNA was about 100 times weaker than the remaining two compounds; the binding of cisplatin was about 30% stronger than oxoplatin. Cisplatin and oxoplatin formed crosslinks with DNA in lymphocytes, whereas the conjugate induced DNA strand breaks. The lesions evoked by cisplatin and oxoplatin were slowly removed, but damage induced by Se Pt was not repaired after 5 h even at a drug concentration of 10 microM. Severe cytotoxic and genotoxic effects exerted by Se-Pt in normal human lymphocytes preclude its intravenous application in cancer therapy. Teratogenesis Carcinog. Mutagen. 20:119-131, 2000. PMID- 10820423 TI - Induction of micronuclei in bone marrow and sperm head abnormalities after combined exposure of mice to low doses of X-rays and acrylamide. AB - People come into contact with chemical and physical agents which are present in the environment and in workplaces. We investigated the effects of combined exposures to low doses of X-rays (0.05-0.25 Gy) and acrylamide (AA; 75 mg/kg bw) in the somatic and germ cells of outbred male mice by using a bone-marrow micronucleus test and a sperm morphology test. Combined treatment of germ cells to 0.25 Gy of X-rays + 75 mg/kg bw of acrylamide enhanced the effect induced by each agent given alone. The results confirmed the sensitivity to damage of spermatozoa and late spermatids, which can be demonstrated by sperm head abnormalities and reduced fertility. The sensitivity of somatic cells to acrylamide alone was similar to that of germ cells. Combined exposure to 0.05 Gy + 75 mg/kg bw of AA induced micronuclei in polychromatic erythrocytes of bone marrow although each dose did not produce a mutagenic effect. Teratogenesis Carcinog. Mutagen. 20:133-140, 2000. PMID- 10820424 TI - Enhanced cytogenetic and antitumor effects by 9-nitrocamptothecin and antineoplastics. AB - Enhanced sister chromatid exchange (SCE) frequency by either melphalan (Mel) or epirubicin (Epir) was observed when human lymphocytes were exposed in vitro to 9 nitro-20(S)-camptothecin (9NC). A correlation was observed between the magnitude of the SCE response and the depression of the cell proliferation index. The antitumor activity of Mel and of 9NC was tested on leukemia P-388-bearing mice. The two chemicals in combination enhance antitumor activity in a synergistic manner. Therefore, the in vivo antitumor effect of Mel in conjunction with 9NC appears to correlate well with the in vitro synergistic effect on SCE induction caused by the combined Mel plus 9NC treatment. Teratogenesis Carcinog. Mutagen. 20:141-146, 2000. PMID- 10820425 TI - 17-alpha-ethinylestradiol and norgestrel in combination induce micronucleus increases and aneuploidy in human lymphocyte and fibroblast cultures. AB - Oral contraceptives are highly efficient and easily administered drugs; however, it must not be forgotten that they are composed of chemical substances which can be classified as potential carcinogens. Testing of a substance for genotoxicity represents a reliable approach both to evaluate the genetic hazard and to obtain information on its possible tumorigenic (cancerogenic) properties. The present study was undertaken to evaluate through carefully planned and controlled investigations the in vitro cytogenetic effects of oral contraceptives (ethynilestradiol and norgestrel mixed in the proportion 1:5) using three different concentrations, with two different durations of treatment (48 and 72 h), on two types of human cells (lymphocytes and fibroblasts) and a series of short-term test procedures: sister chromatid exchange (SCE), micronucleus test (MN), and chromosome aberrations (CA). In addition, the FISH procedure and in vitro anaphase and metaphase preparation analyses were performed. In contrast to CA and SCE frequencies, the frequency of MN in treated blood lymphocytes showed higher values by comparison with the controls, although the difference was statistically significant only for the lowest concentration (P = 0. 016). When using pancentromeric alphoid probes, the FISH procedure gave positive signals in more than 85% of micronuclei, clearly indicating that MN may contain whole chromosomes rather than acentric fragments. Unlike the lymphocytes, the fibroblasts showed dose-dependent effects, although those treated with the highest hormone concentrations showed an increased number of highly damaged cells (cytoplasmatic vacuolization, nuclear fragmentation, etc.), a decreased number of anaphase cells, a large number of which were abnormal, and a reduction of mitotic index. In conclusion, our data confirm that hormones do not induce structural chromosome aberrations in lymphocytes and indicate that ethynilestradiol and norgestrel have an aneugenic effect on fibroblast and lymphocyte cultures; FISH analysis on micronuclei from lymphocyte cultures and anaphase preparations from fibroblast cultures support this hypothesis. Teratogenesis Carcinog. Mutagen. 20:147-159, 2000. PMID- 10820426 TI - Cytogenetic and molecular biomonitoring of agricultural workers exposed to pesticides in Brazil. AB - The use of agricultural chemicals without correct protection may lead to alterations in the genetic material of cells and the possible development of several types of tumors. The individual genetic variability in the enzymes which metabolize agricultural chemicals is also involved in this process, such as when the enzymes are not efficient in the detoxifying process of the organism, the metabolic subproducts accumulate, contributing to the tumorigenic process. Cytogenetic monitoring was carried out on a group of 20 male workers occupationally exposed to a mixture of pesticides in the town of Sao Jeronimo da Serra, PR (Brazil). Student's t = test and Wilcoxon's test showed, respectively, that there was no significant difference between the chromosome aberration frequencies between the exposed and control groups and between the paired individuals. However, there was a significant difference in the two analyses regarding the mitotic index of the sampled individuals. Smoking and time of exposure to agricultural chemicals did not influence the cytogenetic responses obtained, but the mitotic index of the control individuals was higher than that of the exposed individuals from the different age groups. The GSTM1 gene polymorphism was 33% null. When statistical tests were carried out to assess the relationship of the GSTM1 genotypes with the chromosome aberrations and mitotic indexes, there was no significant difference. The CA frequencies found in this study were low, making it difficult to associate it with the GSTM1 gene polymorphism. Teratogenesis Carcinog. Mutagen. 20:161-170, 2000. PMID- 10820427 TI - Schwann cell remyelination is restricted to astrocyte-deficient areas after transplantation into demyelinated adult rat brain. AB - The ability to generate large numbers of Schwann cells from a peripheral nerve biopsy makes them potential candidates for the clinical application of cell transplantation to enhance remyelination in human demyelinating disease. Transplant-derived Schwann cell remyelination has previously been demonstrated in the spinal cord but not for demyelinated axons in the brain, a more likely site for initial clinical intervention. We have transplanted Schwann cells from male neonatal rat sciatic nerves into ethidium bromide-induced areas of demyelination in the deep cerebellar white matter of adult female rats. The extent of Schwann cell remyelination 28 days after transplantation was significantly increased in lesions that received direct injections of Schwann cells compared with non transplanted lesions. Using in situ hybridisation to identify the rat Y chromosome, transplanted male cells were found to co-localise with the P0 immunoreactive area of Schwann cell remyelination. Combined immunohistochemistry and in situ hybridisation confirmed that many remyelinating Schwann cells were transplant-derived. P0 immunoreactivity and transplanted male cells were found in GFAP-negative, astrocyte-free areas. Transplanted Schwann cells were not identified outside of transplanted lesions, nor did they did not contribute to remyelination of a lesion at a distance from the site of transplantation. Our findings indicate that demyelinated axons in the adult brain can be remyelinated by transplanted Schwann cells but that migration and remyelination are restricted to areas from which astrocytes are absent. PMID- 10820428 TI - Differential expression of alternatively spliced neural cell adhesion molecule L1 isoforms during oligodendrocyte maturation. AB - The expression of neural cell adhesion molecules and myelin-specific molecules is precisely regulated according to cell type and developmental age. We investigated whether different isoforms of these molecules change during development of oligodendrocytes. Immature oligodendrocytes cultured from embryonic day 18 rat cerebrum were distinguished into early stage and late stage by morphological and immunocytochemical criteria. mRNA levels of the neural cell adhesion molecule L1 in late-stage immature oligodendrocytes were approximately fivefold higher than in early-stage cells, but early-stage immature oligodendrocytes predominantly expressed an L1 spliced isoform lacking two region (exon 2 and 27). Late-stage cells expressed full-length L1 identical to the neuronal form. mRNA for the neural cell adhesion molecules NCAM and MAG did not show any difference in expression pattern. These results suggest that alternatively spliced isoforms of L1 might be regulated by temporal and spatial factors during oligodendrocyte development. PMID- 10820429 TI - Dimerization-dependent block of the proapoptotic effect of p75(NTR). AB - The biochemical mechanism by which neurons become dependent on neurotrophins for survival is unknown. We found previously that the common neurotrophin receptor, p75(NTR), is a mediator of neurotrophin dependence and that this effect requires a novel type of domain dubbed a neurotrophin dependence domain. We report here that, in contrast to other proapoptotic receptors such as Fas, apoptosis induction by p75(NTR) requires monomerization, with dimerization inhibiting the effect. Blocking the proapoptotic effect of the monomer by dimerization requires a distinct domain that lies at the carboxyterminus of p75(NTR). These results define a novel type of domain required for inhibiting apoptosis induction by p75(NTR). PMID- 10820430 TI - Functional expression of P-glycoprotein and multidrug resistance-associated protein (Mrp1) in primary cultures of rat astrocytes. AB - Although it has been well established that the drug efflux pump P-glycoprotein (P gp) protects the brain against the entry of cytotoxic drugs, its real in situ localization, i.e., at brain capillary endothelial cells or on astrocyte foot processes, is still controversial. The aim of this study was to compare the expression of P-gp and of multidrug resistance-associated protein (Mrp1), another drug efflux pump, in cultured neonatal rat brain astrocytes and in cultured brain capillary endothelial cells. Reverse transcriptase-polymerase chain reaction (RT PCR) analysis showed that the mdr1b gene was preferentially expressed in astrocytes, whereas both mdr1a and mdr1b mRNA were detected in endothelial cells. Moreover, the mrp1 gene encoding Mrp1 was expressed in both cell types. Western blotting analysis revealed higher expression of P-gp in endothelial cells as compared with astrocytes, but higher expression of Mrp1 in astrocytes. Moreover, P-gp and Mrp1 expression was not modified in more differentiated astrocytes obtained when cultured with db-cAMP for 48 hr. Our functional analysis of P-gp showed a modest effect of P-gp modulators (CsA, verapamil, PSC 833) on the uptake of colchicine (a substrate of P-gp) by astrocytes, whereas they increased by about 50% the uptake of vincristine (a common substrate of P-gp and MRP) by astrocytes. MRP modulators (genistein, probenecid, and sulfinpyrazone) did not modify the uptake of colchicine but increased that of vincristine with a major effect found for sulfinpyrazone. Moreover, indomethacin, probenecid, and sulfinpyrazone increased the uptake of fluorescein (a substrate of MRP but not of P-gp). Taken together, our results provide the first biochemical and functional evidence supporting the expression of P-gp and Mrp1 in rat cultured astrocytes. PMID- 10820432 TI - Nitric oxide synthase induction in astroglial cell cultures: effect on heat shock protein 70 synthesis and oxidant/antioxidant balance. AB - Glial cells in the nervous system can produce nitric oxide in response to cytokines. This production is mediated by the inducible isoform of nitric oxide synthase. Radical oxygen species (ROS) and nitric oxide (NO) derivatives have been claimed to play a crucial role in many different processes, both physiological such as neuromodulation, synaptic plasticity, response to glutamate, and pathological such as ischemia and various neurodegenerative disorders. In the present study we investigated the effects of NO synthase (iNOS) induction in astrocyte cultures on the synthesis of heat shock proteins, the activity of respiratory chain complexes and the oxidant/antioxidant balance. Treatment of astrocyte cultures for 18 hr with LPS and INFgamma produced a dose dependent increase of iNOS associated with an increased synthesis of hsp70 stress proteins. This effect was abolished by the NO synthase inhibitor L-NMMA and significantly decreased by addition of SOD/CAT in the medium. Time course experiments showed that iNOS induced protein expression increased significantly by 2 hr after treatment with LPS and INFgamma and reached a plateau at 18 hr; hsp70 protein synthesis peaked around 18 and 36 hr after the same treatment. Addition to astrocytes of the NO donor sodium nitroprusside resulted in a dose dependent increase in hsp70 protein that was comparable to that found after a mild heat shock. Additionally, a decrease in cytochrome oxidase activity, a marked decrease in ATP and protein sulfhydryl contents, an increase in the activity of the antioxidant enzymes mt-SOD and catalase were found which were abolished by L-NMMA. These findings suggest the importance of mitochondrial energy impairment as a critical determinant of the susceptibility of astrocytes to neurotoxic processes and point to a possible pivotal role of hsp70 in the signalling pathways of stress tolerance. PMID- 10820431 TI - Expression of the AMF/neuroleukin receptor in developing and adult brain cerebellum. AB - The peptide sequence of autocrine motility factor (AMF), a tumor secreted cytokine that induces cell motility, corresponds to that of the previously identified cytokine/enzyme, neuroleukin/glucose-6-phosphate isomerase. Neuroleukin is a neurotrophic factor that promotes neuronal survival and sprouting at the neuromuscular junction. The AMF receptor (AMF-R) has been identified and shown to be highly expressed in malignant tumors with minimal expression in adjacent normal tissue. Neuroleukin mRNA is highly expressed in the cerebellum and we therefore undertook a developmental study of AMF-R expression in rat cerebellum. As determined by immunoblot, AMF-R is expressed at equivalent high levels in brain and cerebellum of postnatal day 5 (P5) and 12 (P12) rats and at significantly reduced levels in the adult. Coimmunofluorescence studies with MAP-2 and gamma-actin revealed that at P12, AMF-R was mainly localized to Purkinje and granule cells. Moreover, the premigratory cells of the external granular layer were also immunoreactive for AMF-R suggesting a role for AMF-R in granule cell migration during cerebellar development in the first two weeks after birth. In the adult, AMF-R distribution was similar to P12, although weaker, and was localized to Purkinje and granule cells. AMF-R labeling of GFAP positive glial processes could not be detected in cerebellar sections although in cerebellar primary cultures, both neurons and glial cells were labeled for AMF-R. In neurons, AMF-R labeling was present in the cell body, neurites and growth cones. These data indicate that regulation of the neurotrophic function of neuroleukin might be regulated spatially and temporally by expression of its receptor, AMF-R, in developing and adult cerebellum. PMID- 10820433 TI - Constitutive activity and differential localization of p38alpha and p38beta MAPKs in adult mouse brain. AB - To understand the roles of p38 mitogen-activated protein kinase (p38 MAPK) isoforms in adult mouse brain, in vivo activities and detailed expression patterns of two p38 isoforms, p38alpha and p38beta, were examined by using biochemical and immunohistochemical analyses. The result indicated that the activity of both p38alpha and p38b MAPKs in normal adult mouse brain was remarkably high, and the nuclear pool of the p38 isoforms was primarily responsible for most of the constitutive p38 MAPK activity in brain. Both p38alpha and p38beta were highly expressed in brain areas including cerebral cortex, hippocampus, cerebellum, and few nuclei of the brainstem. At the subcellular level, p38alpha was distributed in dendrites and in cytoplasmic and nuclear regions of cell body of neurons, which is in contrast to p38beta, since p38beta was preferentially expressed in nucleus of neurons. These results suggest that the p38 pathway may play an important role, not only in inflammation and neuronal cell death as previously suggested, but also in normal physiology of adult mouse brain. PMID- 10820434 TI - In vivo glutamine hydrolysis in the formation of extracellular glutamate in the injured rat brain. AB - Hydrolysis of extracellular glutamine as a potential source of increased extracellular glutamate in the quinolinic acid (QUIN)-injured brain of the unanesthetized, free-moving rat was examined by microdialysis and HPLC analysis. Injury was initiated by injection of 100 nmoles of QUIN into the hippocampus. Immediately postinjury or 24 hr postinjury, the injection site was perfused with artificial cerebrospinal fluid + (14)C-glutamine to measure its conversion to (14)C-glutamate. L-trans-pyrrolidine-2,4-dicarboxylate (L-PDC), a glutamate uptake inhibitor, was added to the perfusate to enhance the detection of extracellular (14)C-glutamate. QUIN injury was followed by an immediate increase in extracellular glutamate that persisted 24 hr later. When (14)C-glutamine was added to the perfusate, a significant amount of (14)C-glutamate was recovered, and it was greater following QUIN injury than in control animals (P < 0.001). Up to 32% of the extracellular (14)C-glutamine was converted to (14)C-glutamate following QUIN injury. Considering the high concentration of glutamine normally present in the extracellular fluid, glutamine hydrolysis is a potential and important source for the increase in extracellular glutamate after neuronal injury in vivo. PMID- 10820436 TI - Differential mechanisms of neutrophil and monocyte adhesion on neuroblastoma cells: CD18 and VLA-4 integrins mediate adhesion to SK-N-SH, but not to SK-N-MC cell line. AB - We examined the adhesion of monocytes and polymorphonuclear leukocytes (PMNLs) to the neuroblastoma (NB) cell lines SK-N-SH and SK-N-MC, which have some distinct differentiation characteristics. Monocytes adhered to SK-N-SH and SK-N-MC to the same extent (20 +/- 1.4% and 24 +/- 0.8% of monocytes added). Monocyte adhesion to SK-N-SH but not SK-N-MC was partially inhibited by treating monocytes with a mAb to the CD18 (beta2) integrin chain. The adhesion was further inhibited when monocytes were treated with a combination of mAb to CD18 and VLA-4. Treatment of both NB cell lines with interleukin-1alpha (0.5 ng/ml), tumor necrosis factor alpha (100 U/ml), interferon gamma (200 U/ml), or their combinations increased monocyte adhesion to SK-N-SH and SK-N-MC. With each condition, monocyte adhesion to SK-N-SH was partially blocked by mAb to CD18. The inhibition of adhesion to IL 1alpha- or TNFalpha-treated SK-N-SH cells was greater when the monocytes were treated with mAb to both CD18 and VLA-4. In contrast, monocyte adhesion to IL 1alpha or IFNgamma treated SK-N-MC was only slightly inhibited with a combination of mAb to CD18 + VLA-4 and there was no inhibition at all to TNFalpha-treated SK N-MC. Spontaneous PMNL adhesion to SK-N-SH was almost negligible but increased by treating the cell line with IL-1alpha, TNFalpha, IFNgamma or their combinations. A mAb to CD18 blocked this increase in each case. The pattern of adhesion of PMNLs to SK-N-MC was totally different. PMNL adhesion to unstimulated SK-N-MC was very high (24 +/- 1.3%), was not inhibited by mAb to CD18, and did not increase by stimulating the cell line with IL-1alpha, TNFalpha, IFNgamma or their combinations. Overall, these results suggest two distinct patterns of monocyte and PMNL interaction with neural cells, such as the SK-N-SH and MC cell lines. While monocyte and PMNL adhesion to SK-N-SH is mainly via CD18/VLA-4 or the CD18 mechanisms, respectively, leukocyte adhesion to SK-N-MC is CD18- and VLA-4 independent. Thus, leukocyte-neural cell interactions share some mechanisms common also to leukocyte-endothelium interaction, but there are also unique mechanisms which may be neural cell and differentiation specific. PMID- 10820435 TI - Inhibition of the glutamate-induced K(+) current in identified Onchidium neurons by nitric oxide donors. AB - Nitric oxide (NO) acts as a neurotransmitter and neuromodulator in the nervous system of many vertebrates and invertebrates. The effects of extracellularly applied sodium nitroprusside (SNP) and diethylamine NO (C(2)H(5))(2)N[N(O)NO] Na(+) (DEA/NO), NO donors, on a glutamate (Glu)-induced K(+) current in identified Onchidium neurons were investigated using voltage clamp and pressure ejection techniques. Bath-applied SNP (10 microM) and DEA/NO (5-10 microM) reduced the Glu-induced K(+) current without affecting the resting membrane conductance and holding current. The Glu-induced K(+) current also was inhibited by the focal application of SNP to the neuron somata. The suppressing effects of NO donors were concentration-dependent and completely reversible. Pretreatment with hemoglobin (50 microM), a nitric oxide scavenger, and 1H-[1,2, 4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ; 1 microM), a specific inhibitor of NO stimulated guanylate cyclase, decreased the SNP-induced inhibition of the Glu induced current. Bath-applied 50 microM 3-isobutyl-1-methylxanthine (IBMX), a nonspecific phosphodiesterase inhibitor, or intracellular injection of 1 mM guanosine 3',5'-cyclic monophosphate (cGMP) inhibited the Glu-induced current, mimicking the effect of NO donors. These results demonstrate that SNP and DEA/NO inhibit the Glu-induced K(+) current and that the mechanism of NO inhibition of the Glu-induced current involves cGMP-dependent protein kinase. PMID- 10820437 TI - Human neuroblastoma SH-SY5Y cell line: neurosteroid-producing cell line relying on cytoskeletal organization. AB - Pregnenolone, the precursor of all steroids, is synthesized by CNS structures. The synthesis requires an obligatory step involving cholesterol transport to mitochondrial cytochrome P450-cholesterol side chain cleavage (cytP450scc), although the underlying mechanism(s) are still mostly unknown. We used the human neuroblastoma SH-SY5Y cell line to investigate cytP450scc expression and activity and to establish a role of cytoskeleton in pregnenolone synthesis. Immunocytochemical and biochemical approaches revealed that undifferentiated as well as differentiated cells either by retinoic acid (RA) or phorbol ester 12-O tetradecanoylphorbol-13-acetate (TPA), possess cytP450scc and rapidly synthesize pregnenolone in the presence of a NADPH-generating system. The newly neurosteroid formation by SH-SY5Y cells was increased by 22R-hydroxycholesterol and blocked by the cytP450scc inhibitor, aminoglutethimide. When trilostane was used to inhibit 3beta-hydroxysteroid dehydrogenase catalyzing pregnenolone conversion into progesterone, a higher pregnenolone accumulation occurred in TPA-differentiated cells than in RA-differentiated ones. Although SU 10603, a blocker of 17alpha hydroxylase/c17,20-lyase enzyme involved in DHEA formation from pregnenolone, gave rise to an elevated neurosteroid content only in RA-differentiated cells. No difference in pregnenolone levels was found in undifferentiated cells treated with each inhibitor. Thus, differentiation seems to promote pregnenolone metabolizing enzyme activities that may vary upon phenotypic changes induced by RA or TPA. Treatments of differentiated cells with the microtubule-depolymerizing drug colchicine and the actin microfilament-altering agent cytochalasin D decreased pregnenolone synthesis without affecting cell viability or cytP450scc amount. Addition of the cell-permeant cholesterol analogue 22R-hydroxycholesterol known to elude cholesterol transport systems induced pregnenolone synthesis, however, indicating that perturbations in cytoskeleton likely affect endogenous cholesterol transport. The relevance of this finding may rest on the observed involvement of cytoskeletal organization in such events as neuronal plasticity, cognitive function and also neurodegenerative disorders in which neurosteroids have been shown to have a part. PMID- 10820439 TI - Phospholipid incorporation and metabolic conversion of n-3 polyunsaturated fatty acids in the Y79 retinoblastoma cell line. AB - The metabolic conversion of n-3 fatty acids was studied in the human Y79 retinoblastoma cell line. Cultured cells were exposed to increasing concentrations of either 18:3n-3, 22:5n-3, or 22:6n-3, and their phospholipid fatty acid composition was analyzed after 72 hr. Cells internalized the supplemental fatty acids and proceeded to their metabolic conversion. Supplemental 22:6n-3 was directly esterified into cell phospholipids, at levels typical for normal neural retinas (41% by weight of phosphatidylethanolamine fatty acids, and 24% of phosphatidylcholine fatty acids). In contrast, 18:3n-3 was mainly converted to 20:5n-3 and 22:5n-3, both of which appeared in cell phospholipids after exposure to low external concentrations of 18:3n-3 (10 microg/ml). Y79 cells can proceed to the metabolic conversion of 18:3n-3 through elongation and Delta6- and Delta5-desaturation. When cells were exposed to high external concentrations of 18:3n-3 (30 microg/ml), the supplemental fatty acid was directly incorporated, and its relative content increased in both phospholipid classes to the detriment of all other n-3 fatty acids. Cells cultured in the presence of 22:5n-3 did not incorporate 22:6n-3 into their phospholipids but did incorporate 20:5n-3 and 22:5n-3. The data suggest that Y79 cells can proceed to the microsomal steps of n-3 metabolism, involving elongation, desaturation, and chain shortening of 22C fatty acids. Although Y79 cells avidly used supplemental 22:6n-3 for phospholipid incorporation at levels typical for normal photoreceptor cells, they failed to match such levels through metabolic conversion of n-3 parent fatty acids. The terminal step of the very long-chain polyunsaturated fatty acid synthesis, consisting in Delta6 desaturation followed by peroxisomal chain shortening of 24C-fatty acids, could be rate-limiting in Y79 cells. PMID- 10820438 TI - Near-terminus axonal structure and function following rat sciatic nerve regeneration through a collagen-GAG matrix in a ten-millimeter gap. AB - The objectives of this study were to evaluate the regenerated axon structure at near-terminal locations in the peroneal and tibial branches 1 year following implantation of several tubular devices in a 10-mm gap in the adult rat sciatic nerve and to determine the extent of recovery of selected sensory and motor functions. The devices were collagen and silicone tubes implanted alone or filled with a porous collagen-glycosaminoglycan matrix. Intact contralateral nerves and autografts were used as controls. Nerves were retrieved at 30 and 60 weeks postoperatively for histological evaluation of the number and diameter of regenerated axons proximal and distal to the gap and in the tibial and peroneal nerve branches, near the termination point. Several functional evaluation methods were employed: gait analysis, pinch test, muscle circumference, and response to electrical stimulation. A notable finding was that the matrix-filled collagen tube group had a significantly greater number of large-diameter myelinated axons (> or =6 microm in diameter) in the distal nerve branches than any other group, including the autograft group. These results were consistent with previously reported electrophysiological measurements that showed that the action potential amplitude for the A fibers in the matrix-filled collagen tube group was greater than for the autograft control group. Functional testing revealed the existence of both sensory and motor recovery following peripheral nerve regeneration through all devices; however, the tests employed in this study did not show differences among the groups with regeneration. Electrical stimulation in vivo showed that threshold parameters to elicit muscle twitch were the same for reinnervating and control nerves. The investigation is of importance in showing for the first time the superiority of a specific fully resorbable off-the-shelf device over an autograft for bridging gaps in peripheral nerve, with respect to the near-terminus axonal structure. PMID- 10820442 TI - Dead Ends and Detours En Route to Total Syntheses of the 1990s A list of abbreviations can be found at the end of the article. AB - From the very beginning organic chemistry and total synthesis have been intimately joined. In fact, one of the first things that freshmen in organic chemistry learn is how to join two molecules together to obtain a more complex one. Of course they still have a long way to go to become fully mature synthetic chemists, but they must have the primary instinct to build molecules, as synthesis is the essence of organic chemistry. With the different points of view that actually coexist in the chemical community about the maturity of the science (art, or both) of organic synthesis, it is clear that nowadays we know how to make almost all of the most complex molecules ever isolated. The primary question is how easy is it to accomplish? For the readers of papers describing the total synthesis of either simple or complex molecules, it appears that the routes followed are, most of the time, smooth and free of troubles. The synthetic scheme written on paper is, apparently, done in the laboratory with few, if any, modifications and these, essentially, seem to be based on finding the optimal experimental conditions to effect the desired reaction. Failures in the planned synthetic scheme to achieve the goal, detours imposed by unexpected reactivity, or the absence of reactivity are almost never discussed, since they may diminish the value of the work reported. This review attempts to look at total synthesis from a different side; it will focus on troubles found during the synthetic work that cause detours from the original synthetic plan, or on the dead ends that eventually may force redesign. From there, the evolution from the original route to the final successful one that achieves the synthetic target will be presented. The syntheses discussed in this paper have been selected because they contain explicit information about the failures of the original synthetic plan, together with the evolution of the final route to the target molecule. Therefore, they contain a lot of useful negative information that may otherwise be lost. PMID- 10820440 TI - Glutamate and gamma-aminobutyric acid content and release of synaptosomes from temporal lobe epilepsy patients. AB - During surgical intervention in medically refractory temporal lobe epilepsy (TLE) patients, diagnosed with either mesial temporal lobe sclerosis (MTS)- or tumor (T)-associated TLE, biopsies were taken from the anterior temporal neocortex and the hippocampal region. Synaptosomes, isolated from these biopsies were used to study intrasynaptosomal Ca(2+) levels ([Ca(2+)](i)), and glutamate and gamma aminobutyric acid (GABA) contents and release. All synaptosomal preparations demonstrated a basal [Ca(2+)](i) of about 200 nM, except neocortical synaptosomes from MTS-associated TLE patients (420 nM). K(+)-induced depolarization resulted in a robust increase of the basal [Ca(2+)](i) in all preparations. Neocortical synaptosomes from TLE patients contained 22.9 +/- 3.0 nmol glutamate and 4.6 +/- 0.5 nmol GABA per milligram synaptosomal protein, whereas rat cortical synaptosomes contained twice as much glutamate and four times as much GABA. Hippocampal synaptosomes from MTS-associated TLE patients, unlike those from T associated TLE patients, contained about 70% less glutamate and 55% less GABA than neocortical synaptosomes. Expressed as percentage of total synaptosomal content, synaptosomes from MTS-associated TLE patients exhibited an increased basal and a reduced K(+)-induced glutamate and GABA release compared to rat cortical synaptosomes. In MTS-associated TLE patients, only GABA release from neocortical synaptosomes was partially Ca(2+)-dependent. Control experiments in rat synaptosomes demonstrated that at least part of the reduction in K(+)-induced release can be ascribed to resection-induced hypoxia in biopsies. Thus, synaptosomes from MTS-associated TLE patients exhibit a significant K(+)-induced increase in [Ca(2+)](i), but the consequent release of glutamate and GABA is severely impaired. Our data show that at least part of the differences in glutamate and GABA content and release between human biopsy material and fresh rat tissue is due to the resection time. PMID- 10820444 TI - Atomic Glimpses on a Billion-Year-Old Molecular Machine. PMID- 10820443 TI - The Orbital Origins of Magnetism: From Atoms to Molecules to Ferromagnetic Alloys. AB - A chemical view of spin magnetic phenomena in finite (atoms and molecules) and infinite (transition metals and their alloys) systems using the concepts of bonding and electronic shielding is presented. The concept is intended to serve as a semiquantitative signpost for the synthesis of new ferromagnets. After a concise overview of the historic development of related theories developed within the physics community, the consequences of spin-spin coupling (made manifest in the exchange or Fermi hole) in atoms and molecules are explored. Upon moving to a paramagnetic state, the majority/minority spin species become more/less tightly bound to the nucleus, resulting in differences in the energies and spatial extents of the two sets of spin orbitals. By extrapolating well-known arguments from ligand-field theory, the paucity of ferromagnetic transition metals arises from quenching the paramagnetism of the free atoms due to strong interatomic interactions in the solid state. Critical valence electron concentrations in Fe, Co, and Ni, however, result in local electronic instabilities due to the population of antibonding states at the Fermi level varepsilon(F). Removal of these antibonding states from the vicinity of varepsilon(F) is the origin of ferromagnetism; in the pure metals this results in strengthening the chemical bonds. In the 4d and 5d transition metals, the valence d orbitals are too well shielded from the nucleus, so a transition to a ferromagnetic state does not result in sufficiently large changes to occur. Thus, the exceptional occurence of ferromagnetism only in the first transition series appears to parallel the special main-group chemistry of the first long period. A connection between ferromagnetism in the transition metals and Pearson's absolute hardness eta is easily established and shows that ferromagnetism appears only when eta<0.2 eV in the nonmagnetic calculation. As expected from the principle of maximum hardness, Fe, Co, and Ni all become harder upon moving to the more stable ferromagnetic state. Magnetism in intermetallic alloys follows the same path. Whether or not an alloy contains ferromagnetic elements, the presence of antibonding states at varepsilon(F) serves as a "fingerprint" to indicate a ferromagnetic instability. The differences in the sizes of the local magnetic moments on the constituent atoms of a ferromagnetic alloy can be understood in terms of the relative contributions to the density of states at varepsilon(F) in the nonmagnetic calculations. Appropriately parameterized, nonmagnetic, semi-empirical calculations can also be used to expose the ferromagnetic instability in elements and alloys. These techniques, which have become relatively commonplace, can be used to guide the synthetic chemist in search of new ferromagnetic materials. PMID- 10820445 TI - How Does Tyrosinase Work? Recent Insights from Model Chemistry and Structural Biology This work was supported by the Medicine and Science Center of the University of Mainz (H.D.) and the Deutsche Forschungsgemeinschaft (F.T., R.D.). The authors thank M.Moller for help with the graphical artwork. PMID- 10820446 TI - From Molecular Flat Tapers, Discs, and Cones to Supramolecular Cylinders and Spheres using Frechet-Type Monodendrons Modified on their Periphery Financial support by the National Science Foundation (DMR-97-08581), the British Engineering and Physical Science Research Council (UK), and the Synchrotron Radiation Source at Daresbury (UK) is gratefully acknowledged. PMID- 10820447 TI - Nickel-Catalyzed Cross-Coupling of Aryl Chlorides with Aryl Grignard Reagents N Heterocyclic Carbenes, Part 27. This work was supported by Aventis R&T, the Deutsche Forschungsgemeinschaft (DFG), the Bayerischer Forschungsverbund Katalyse (FORKAT), Degussa-Huls AG, and the Fonds der Chemischen Industrie (studentships for V.P.W.B. and T.W.). We thank C. Brossmer, K. Denk, R. W. Eckl, M. G. Gardiner, J. Schwarz, and C.-P. Reisinger for preparation of commercially unavailable ligands used in the screening assay and T. Boussie from Symyx Technologies for helpful discussions. Part 26: V. P. W. Bohm, C. W. K. Gstottmayr, T. Weskamp, W. A. Herrmann, J. Organomet. Chem. 2000, 595, 186-190. PMID- 10820448 TI - High-Spin Molecules: A Novel Cyano-Bridged MnMo Molecular Cluster with a S=51/2 Ground State and Ferromagnetic Intercluster Ordering at Low Temperatures Financial support by the Swiss National Science Foundation through Project No. 21 52699.97 and the TMR Research Network ERBFMRXCT 980181 of the European Union, entitled: "Molecular Magnetism: From Materials towards Devices" is gratefully acknowledged. The authors also thank Thierry Strassle from the Paul Scherrer Institute, Villigen, for assistance with the heat capacity measurement and Professor Fernando Palacio, University of Zaragoza, for stimulating discussions. PMID- 10820449 TI - Reaction of Dichloromethyltris(trimethylsilyl)silane with Organolithium Reagents: Synthesis of an Intramolecularly Donor-Stabilized Silene This work was supported by the State of Mecklenburg-Vorpommern, by the Deutsche Forschungsgemeinschaft, and by the Fonds der Chemischen Industrie. We thank Prof. M. Michalik and Prof. N. Stoll for recording NMR and mass spectra. PMID- 10820450 TI - Linking Icosahedral, Strong Molecular Magnets {MoFe} to Layers-A Solid-State Reaction at Room Temperature The authors thank M. Baumann, C. Kuhlmann, and S. Q. N. Shah for their assistance, and Prof. A. X. Trautwein and Dr. V. Schunemann (University of Lubeck) for the measurement of the (57)Fe Mossbauer spectrum. S.S. (Department of Chemistry, IIT Kanpur, India) thanks the Alexander von Humboldt Foundation for financing his three-month stay in Bielefeld. PMID- 10820451 TI - Thirty Electrons "Trapped" in a Spherical Matrix: A Molybdenum Oxide-Based Nanostructured Keplerate Reduced by 36 Electrons. PMID- 10820452 TI - A Molecular Knot with Twelve Amide Groups-One-Step Synthesis, Crystal Structure, Chirality Extracts were presented during a talk at the Universitat Dusseldorf on January 11, 2000. We thank Dr. Christian Seel and Dr. Rudolf Hartmann for suggestions and measurements. PMID- 10820453 TI - Highly Regioselective Oxygenation of C-H Bonds: Diamidomanganese Constructs with Attached Substrates as Catalyst Models Financial support has been provided by Columbia University and the Camille and Henry Dreyfus Foundation. P.M.W. acknowledges the REU-NSF program for summer support. We also thank Dr. J. B. Schwarz (manuscript preparation) and Dr. Y. Itagaki (mass spectrometry). PMID- 10820454 TI - Solid-Phase Synthesis of Unprotected N-Glycopeptide Building Blocks for SPOT Synthesis of N-linked Glycopeptides. PMID- 10820455 TI - Can a Homometallic Chain Be Ferrimagnetic? This research was supported by CICYT (Grant PB96/0163) and OENB (Grants 6630 and 7967). F.A.M. thanks Prof. C. Kratky and Dr. F. Belaj (Universitat Graz) for the use of experimental equipment. A.E. and R.V. thanks Prof. Dante Gatteschi (University degli Studi di Firenze) for helpful discussions. PMID- 10820456 TI - Stereoselectivity in the Formation of a Cyclic Trinuclear cis-Dioxomolybdenum(VI) Complex of a Chiral Siderophore Analogue This work was supported by the Fonds der Chemischen Industrie. We thank Dr. T. A. Dransfield, York, for the ES mass spectrum and Dr. T. Vogel and B. Beermann, Munster, for recording the CD spectra. PMID- 10820457 TI - An Enzyme-Labile Safety Catch Linker for Combinatorial Synthesis on a Soluble Polymeric Support This work was supported by the Bundesministerium fur Bildung und Forschung and BASF AG. PMID- 10820458 TI - The First Structurally Characterized Metal Complex with the Molecular Unit M=C=C=C=CR(2) This work was supported by the Deutsche Forschungsgemeinschaft (SFB 347), the Fonds der Chemischen Industrie (Ph. D. grant to K. I.), and the BASF AG. PMID- 10820459 TI - Homoleptic Carbonyl Ni-Pd Clusters: Synthesis of PMID- 10820460 TI - [Li] PMID- 10820461 TI - Novel Oxyfunctionalized Phosphonite Ligands for the Hydroformylation of Isomeric n-Olefins. PMID- 10820462 TI - First Noncovalently Bound Calix PMID- 10820463 TI - Lithium Amides: Intra-Aggregate Complexation of Lithium and Entropy Control of Basicity. PMID- 10820464 TI - Synthesis, Crystal Structure, and Magnetic Behavior of Linear MU(IV) Complexes (M=Co, Ni, Cu, Zn). PMID- 10820465 TI - A Catalytic Asymmetric Strecker-Type Reaction: Interesting Reactivity Difference between TMSCN and HCN This work was supported by CREST and RFTF. We thank Professor Kobayashi and Dr. Ishitani at the University of Tokyo for kindly showing us their recent results and the procedure for the preparation of HCN. We also thank Professor Hoveyda in Boston College for kindly showing us his recent results. PMID- 10820466 TI - Metric Engineering of Crystalline Inclusion Compounds by Structural Mimicry This work was supported in part by the MRSEC Program of the National Science Foundation (Award Number DMR-9809364) and by the Natural Sciences and Engineering Research Council of Canada (postdoctoral fellowship for K.T.H.). PMID- 10820467 TI - The Eight Stereoisomers of LNA (Locked Nucleic Acid): A Remarkable Family of Strong RNA Binding Molecules We acknowledge the Danish Natural Science Research Council, the Danish Technical Research Council, and Exiqon A/S for financial support. Ms Britta M. Dahl is thanked for oligonucleotide synthesis, Dr. Carl E. Olsen for MALDI-MS analysis, and Ms. Karen Jorgensen for recording CD spectra. PMID- 10820468 TI - Metal Incorporation into and Dimerization of M(3)E(4) Clusters (M=Mo, W; E=S, Se) in Supramolecular Assemblies with Cucurbituril: A Molecular Model of Intercalation This work was supported by the Russian Foundation for Basic Research (research grant 99-03-32788) and an EU INTAS collaboration (research grant 96-1256). A.V.V. thanks the Royal Society for financial supporting of his visit to the University of Newcastle upon Tyne. We also thank the EPSRC (UK) for funding. PMID- 10820469 TI - Nanoscale Super Clusters of Clusters Assembled around a Dendritic Core We thank the Royal Society (Smithson Research Fellowship at Peterhouse to D.S.S.), The Singapore Government (P.G.G.), and ICI Wilton for financial support, and Dr. S. J. Teat at the Daresbury SRS (UK) for assistance in crystallography. PMID- 10820470 TI - The Synthesis of Streptogramin Antibiotics: (-)-Griseoviridin and Its C-8 Epimer Financial support has been provided by the National Institutes of Health (NIH). We thank the NIH for financial support of this study and Drs. Russell Linderman, Donald Walker, Ronald Spohn, and Enrico Marcantoni for their assistance in various stages of this effort. PMID- 10820471 TI - A Structural Model for the Galactose Oxidase Active Site which Shows Counteranion Dependent Phenoxyl Radical Formation by Disproportionation This work was supported by Grants-in-Aid for Scientific Research (No. 09304062 and 0149219 (Priority Areas) to O.Y. and No. 07CE2004(COE) to A.O.) from the Ministry of Education, Science, Sports, and Culture of Japan, for which we express our thanks. PMID- 10820472 TI - Reaction of Organic Selenocyanates with Hydroxides: The One-Pot Synthesis of Dialkyl Diselenides from Alkyl Bromides. PMID- 10820473 TI - Eliminative Ring Opening of Oxiranium Ions in the Gas Phase This work was supported by the Ministero della Universita e della Ricerca Scientifica e Tecnologica (MURST) and the Consiglio Nazionale delle Ricerche (CNR). Thanks are due to Claudio Lillocci, who kindly provided us with a purified sample of threo-2 dimethylamino-3-methoxybutane. PMID- 10820474 TI - High Oxidation State Organocobalt Complexes: Synthesis and Characterization of Dihydridodisilyl Cobalt(V) Species We thank the National Institutes of Health (Grant GM 28938) for financial support. C.P.L. thanks the Fonds der Chemischen Industrie, Germany, for a Kekule fellowship. Allocation of computing time by the North Carolina Supercomputing Center is gratefully acknowledged. PMID- 10820475 TI - Traceless, Solid-Phase Synthesis of Biarylmethane Structures through Pd-Catalyzed Release of Supported Benzylsulfonium Salts We thank Rhone Poulenc Rorer and Rhone Poulenc Agro for financial support. PMID- 10820476 TI - A Highly Soluble Luminescent Decanuclear Gold(I) Complex with a Propeller-Shaped Structure V.W.-W.Y. acknowledges financial support from the Research Grants Council and The University of Hong Kong. E.C.-C.C. acknowledges the receipt of a postgraduate studentship (1997-1999) and a Croucher Foundation Scholarship (1999 2000), administered by The University of Hong Kong and the Croucher Foundation, respectively. PMID- 10820477 TI - Metal-Mediated Transfer of Electrons between Two Different C-C Single Bonds That Function as Electron-Donor and Electron-Acceptor Units This work was supported by the Fonds National Suisse de la Recherche Scientifique (Grant No. 20-53336.98) and Action COST D9 (European Program for Scientific Research, OFES No. C98.008). PMID- 10820478 TI - An Octanuclear Fe(III) Compound Featuring a New Type of Double Butterfly Iron-Oxo Core Financial support from Sustainable Technologies Australia Ltd. and Postgraduate Awards to P.A. and C.M.K. are gratefully acknowledged. Kevin Berry is thanked for assistance with the magnetic calculations. PMID- 10820479 TI - FBeNg(+) (Ng=He, Ne, Ar): Suitable Cations for Salts of the Lightest Noble Gases? This work was supported by the Italian Ministero dell' Universita e della Ricerca Scientifica e Tecnologica (MURST) and the Consiglio Nazionale delle Ricerche (CNR). PMID- 10820480 TI - Self-Assembly of Rotaxane-Like Complexes with Macrocycles Containing Reversible Coordinate Bonds This work was supported by the Korea Science and Engineering Foundation (1999-2-123-001-3). We thank Professor Moon-Gun Choi for help in X-ray crystallography, and Professor Weontae Lee for assistance with EXSY experiments. PMID- 10820481 TI - Silole-Thiophene Alternating Copolymers with Narrow Band Gaps This work was partly supported by a Grant-in-Aid (No. 09239103) from the Ministry of Education, Science, Sports and Culture, Japan, and Japan Chemical Innovation Institute. PMID- 10820482 TI - Telomerase and telomere stability: a new class of tumor suppressor? AB - Introduction of telomerase into normal cells provides telomere maintenance and an extended cellular life span, establishing the critical role of telomere attrition in cellular senescence. Additional data surrounding this observation suggest that expression of telomerase renders these "mortal" cells genomically stable with decreased frequencies of mutation, ultimately leading to continued proliferation without signs of changes typically associated with progression to a cancer-like phenotype. Interestingly, oncogenic insult after exogenous telomerase expression does not result in cellular transformation, yet addition of an oncogene first followed by telomerase does transform cells. Taken together, these results imply that order of addition is important for telomerase-mediated genomic protection and that telomerase expression is critical for the transformation process. The hypothesis proposed here is that telomerase, via its function in telomere stabilization, is capable of protecting cells from acquiring the required mutations and genomic instability necessary for malignant transformation, suggesting that telomerase is not an oncogene but may act as a novel class of tumor suppressor. PMID- 10820483 TI - Eyes wide shut: protein kinase C isozymes are not the only receptors for the phorbol ester tumor promoters. AB - In addition to the well-characterized interaction with classical and novel protein kinase C (PKC) isozymes, the phorbol ester tumor promoters bind to other receptors lacking kinase activity. Among these novel phorbol ester receptors, two families of proteins may play a role in the regulation of cell growth and malignant transformation: chimaerins and ras guanyl-releasing protein (ras-GRP). These proteins possess a single copy of the C1 domain that is involved in binding of phorbol esters and the lipid second messenger diacylglycerol. Four isoforms of chimaerins (alpha1-, alpha2-, beta1-, and beta2-chimaerins) have been isolated to date, all of them possessing GTPase-activating protein activity for Rac, a small GTP-binding protein that controls actin cytoskeleton organization, cell-cycle progression, adhesion, and migration. Ras-GRP is a guanine nucleotide exchange factor for ras and promotes malignant transformation in fibroblasts in a phorbol ester-dependent manner. The C1 domain in Ras-GRP may, therefore, have a dominant role in Ras-GRP activation and is essential for phorbol ester-dependent activation of downstream effectors of ras, i.e., the mitogen-activated protein kinase cascade. Thus, a novel concept emerges in which phorbol esters may exert cellular responses through pathways not involving phorbol ester-responsive PKC isozymes. The discovery of "nonPKC" phorbol ester receptors adds an additional level of complexity to the understanding of phorbol ester effects and the molecular mechanisms of carcinogenesis. PMID- 10820484 TI - Development of a prostate cDNA microarray and statistical gene expression analysis package. AB - A cDNA microarray comprising 5184 different cDNAs spotted onto nylon membrane filters was developed for prostate gene expression studies. The clones used for arraying were identified by cluster analysis of > 35 000 prostate cDNA library derived expressed sequence tags (ESTs) present in the dbEST database maintained by the National Center for Biotechnology Information. Total RNA from two cell lines, prostate line 8.4 and melanoma line UACC903, was used to make radiolabeled probe for filter hybridizations. The absolute intensity of each individual cDNA spot was determined by phosphorimager scanning and evaluated by a bioinformatics package developed specifically for analysis of cDNA microarray experimentation. Results indicated 89% of the genes showed intensity levels above background in prostate cells compared with only 28% in melanoma cells. Replicate probe preparations yielded results with correlation values ranging from r = 0.90 to 0.93 and coefficient of variation ranging from 16 to 28%. Findings indicate that among others, the keratin 5 and vimentin genes were differentially expressed between these two divergent cell lines. Follow-up northern blot analysis verified these two expression changes, thereby demonstrating the reliability of this system. We report the development of a cDNA microarray system that is sensitive and reliable, demonstrates a low degree of variability, and is capable of determining verifiable gene expression differences between two distinct human cell lines. This system will prove useful for differential gene expression analysis in prostate-derived cells and tissue. PMID- 10820485 TI - A connexin 43 mutant lacking the carboxyl cytoplasmic domain inhibits both growth and motility of mouse 3T3 fibroblasts. AB - Connexins have been shown to inhibit the growth of a wide number of communication deficient cells both in vivo and in vitro, but the molecular mechanism remains largely unknown. In previous work we have shown that stable transfectants of 3T3 A31 fibroblasts, which express a Connexin 43 (Cx43) mutant (Cx43-256M) consisting of amino acids 1-256 of rat Cx43 fused to a c-myc tag, exhibit a decreased basal growth rate and weakened mitogenic response to platelet derived growth factor compared with either the parent cell line or cells transfected with an expression vector that did not encode a functional protein. Here we have investigated further the growth characteristics of these cells in order to establish the mechanism by which this protein suppresses cell growth. Analysis of DNA synthesis in individual cells by immunofluorescence staining of bromodeoxyuridine incorporation demonstrated that the slow growth of Cx43-256M cells was due to a decrease in the number of cells that undergo DNA synthesis following growth factor stimulation. This was associated with an increased proportion of the cell population in the G2/M phases of the cell cycle suggesting growth may be arrested during G2 or metaphase. In addition to effects on cell growth, Cx43-256M expression inhibited cell motility as assayed both in a wounding assay and in a Boyden chamber assay. These results now raise the question as to whether the primary effect of the Cx43-256M protein is on cell growth or cell motility. PMID- 10820486 TI - The role of wild-type and mutated N-ras in the malignant transformation of liver cells. AB - In order to determine the role of N-ras overexpression and mutation in malignant liver cell transformation, wild-type and mutated N-ras were transfected into the rat liver epithelial cell line OC/CDE 22, and N-ras expression, growth kinetics, growth in soft agar, and tumorigenicity in vivo as well as the involvement of the mitogen-activated protein kinase (MAPK) signal transduction pathway in the expression of the malignant phenotype were analyzed. Although OC/CDE 22 cells transfected with wild-type N-ras showed a high expression of N-ras at the mRNA and protein levels, the cells did not grow in soft agar and were not tumorigenic in vivo. In contrast, OC/CDE 22 cells transfected with mutated N-ras showed anchorage-independent growth and were tumorigenic. When cultured in fetal bovine serum-supplemented medium, OC/CDE 22 cells expressing mutant N-ras showed a higher proliferation rate than nontransfected OC/CDE 22 cells or OC/CDE 22 cells transfected with wild-type N-ras. When held in serum-free medium, untreated OC/CDE 22 cells did not grow at all, while OC/CDE 22 cells transfected with wild type or mutant N-ras proliferated at a similar rate, which can be explained by the high MAPK activity in these cells. Selective inhibition of the MAPK cascade abolished the growth of OC/CDE 22 cells carrying mutant N-ras in soft agar; furthermore, these cells ceased pile up and formed monolayers on Petri dishes. Thus, activation of the MAPK signaling pathway, though alone not sufficient to malignantly transform liver cells (as shown in liver cells overexpressing wild type N-ras), is not only essential for growth control but also for the expression of the malignant phenotype (as demonstrated in liver cells transformed by mutated N-ras). PMID- 10820487 TI - Transcriptional regulation of human papillomavirus type 16 LCR by different C/EBPbeta isoforms. AB - During genital human papillomavirus (HPV) infection several cytokines are released, such as interleukin-1 (IL-1), tumor necrosis factoralpha (TNFalpha), IL 6, and IL-8. These cytokines may play a role in the immune surveillance against viral infection. Two of these cytokines, IL-1 and TNFalpha, suppress the transcription of the HPV16 early genes. CAATT/ enhancer binding protein, (C/EBPbeta), which is activated by IL-1 and TNFalpha, has been suggested to act as a mediator of this transcriptional downregulation. C/EBPbeta contains three different translation initiation sites that can lead probably by leaky ribosome scanning to the generation of three isoforms of C/EBPbeta, namely full-length C/EBPbeta, liver enriched transcriptional activator protein (LAP), and liver enriched inhibitory protein (LIP). When transiently expressed in C33A and HeLa cells, the first two C/EBPbeta isoforms activate the HPV16 long control region (LCR). LIP, which acts as an antagonist of C/EBPbeta, represses the HPV16 LCR activity. Our observation that treatment of HeLa cells with IL-1 leads to induction of LIP supports the hypothesis that the LCR downregulation by IL-1 is mediated by LIP. PMID- 10820488 TI - Genome-wide loss of heterozygosity analysis of chemically induced rat hepatocellular carcinomas reveals elevated frequency of allelic imbalances on chromosomes 1, 6, 8, 11, 15, 17, and 20. AB - Neoplastic development is a multistep process that involves the stochastic accumulation of heritable genetic alterations in proto-oncogenes, DNA repair genes, and tumor suppressor genes. Loss of heterozygosity (LOH) analysis has been used successfully to identify the genetic determinants of neoplastic development, including tumor suppressor genes, in several species and organs but not in the rat liver. We report the results of a sensitive genome-wide LOH analysis of rat hepatocellular carcinomas (HCCs). Heterozygous rats (Wistar-Furth x Fisher 344) were subjected to an Initiation-Promotion-Progression (IPP) protocol of hepatocarcinogenesis. Two weeks after initiation (by partial hepatectomy, 10 mg/kg diethylnitrosamine), the rats were placed on a diet containing 0.05% phenobarbital (PB). After 24 wk of PB promotion, the rats received either 100 or 1 50 mg/kg ethylnitrosourea. Hepatocellular tumors were resected after a total of 76wk of PB promotion. LOH analysis was completed on 26 HCCs by using 60 microsatellite markers covering all 20 rat autosomes and chromosome X. While 85% of the HCCs had one or more allelic imbalances, the average HCC had 3.3 allelic imbalances (range 0-9). A conditional hypothesis-testing method called the Hot Cold model was used to determine the location of statistically significant elevations in the frequency of allelic imbalances. Elevated allelic imbalances were observed on chromosomes 1q, 6, 8, 11, 15, 17, and 20p. Together, these allelic imbalances suggest that the retinoblastoma and insulin-like growth factor genes as well as the resistance to chemical carcinogenesis (rcc) locus may be involved in HCC development in the rat but that LOH of the p53 gene is not. The elevated rate of allelic imbalances on chromosomes 8,11, and 17 may indicate the location of undiscovered tumor suppressor genes important to neoplastic development in rat liver. Microdissection-based LOH analysis of HCC revealed that contamination of non-neoplastic and nonhepatocellular tissue was not masking LOH in the whole-tumor analysis. There were no statistically significant differences in the frequency of allelic imbalances between HCC of any differentiation state (histological grade). To the degree that it does not reflect differences in etiological factors, the absence of allelic imbalances in chromosomal regions containing the p53 and mamose-6-phosphate/insulin-like growth factor II receptor tumor suppressor genes and the generally low frequency of allelic imbalances in these tumors, suggests that LOH and allelic imbalances play a less significant role in the molecular pathogenesis of HCC in rats than humans. PMID- 10820489 TI - The human PCPH proto-oncogene: cDNA identification, primary structure, chromosomal mapping, and expression in normal and tumor cells. PMID- 10820516 TI - Functional properties of the intestinal wall: novel aspects and recent avenues. PMID- 10820517 TI - Roles of molecular chaperones in pancreatic secretion and their involvement in intestinal absorption. AB - This review focuses on the contribution of molecular chaperones in the secretory process of digestive enzymes and their interaction with enterocytes. By using biochemistry and immunocytochemistry, we have shown that Grp94, Cpn10, Cpn60, and protein disulfide isomerase (PDI) are present all along the rough endoplasmic reticulum-Golgi-granule secretory pathway of the pancreatic acinar cells and are secreted into the acinar lumen. Two other molecular chaperones, Grp78 and the Hsp70, appear to be restricted to the rough endoplasmic reticulum and the trans Golgi apparatus, respectively. We have found that chaperones can be associated with pancreatic enzymes along the secretory pathway. Indeed, double immunogold and immunocoprecipitation revealed an association between Cpn60 and the colipase dependent lipase (CDL) and between Grp94 and the bile salt-dependent lipase (BSDL). These complexes are secreted into the acinar lumen and diverted to the duodenal lumen. These findings led us to investigate these enzyme-chaperone complexes in intestinal tissue. Grp94, Cpn60, and PDI are present on microvilli and on the endosomal compartment of enterocytes. Furthermore, we have shown that the Grp94-BSDL complexes are internalized by enterocytes through classical endocytosis. Upon dissociation of the BSDL-Grp94 complex in the late endosome, BSDL is transferred to the basolateral membrane. We propose that Grp94 interacts with specific receptors and/or could force the associated protein to adopt a specific conformation that allows its binding to corresponding membrane receptors and its internalization by enterocytes. These two hypotheses need not to be exclusive. The existence of such a pancreatic secretion-intestinal absorption link speaks in favor of a coordinated functional connection between these two entities, through molecular chaperones, in order to optimize intestinal activities. PMID- 10820518 TI - Intestinal absorption of peptides through the enterocytes. AB - Transport of intact peptides and proteins from the intestinal lumen into the blood is a unique phenomenon, which differs from the regular process of food digestion and absorption. Intestinal absorption of minute amounts of proteins is, however, being considered as a normal physiological process. It is thus important to define and understand the routes for protein transfer from the intestinal lumen to the blood and the mechanisms by which the macromolecules overcome the sieving barrier of the intestinal wall. The study on insulin has demonstrated that, upon proper introduction into the intestinal lumen, insulin is absorbed by the epithelial cells and transferred to the circulation. The peptides absorbed and transferred to the blood retained their biological activity and induced significant lowering of blood glucose levels. The efficiency of the absorption does not differ among the ileum, duodenum, and colon. Morphological examination demonstrated no alteration of the structural integrity of the epithelia, the enterocytes stay intact with well-developed microvilli, and the cells remain joined by tightly closed junctions. Application of immunocytochemistry on thin tissue sections revealed insulin antigenic sites at different locations depending on the time point. Insulin detected in the lumen of the intestinal tract is absorbed through the endosomal compartment of the epithelial cells rather than passing between cells. Internalization occurs through invaginations of the luminal plasma membrane and vesicular structures of the endosomal compartment. In 5-10 minutes, insulin is transferred to the basolateral membrane and released into the interstitial space to reach the circulation. Definition of the transcytotic pathway will contribute to a better understanding of drug delivery for potential therapeutic applications. PMID- 10820519 TI - Epithelial cell polarity as reflected in enterocytes. AB - Absorptive cells are the main cells present in the intestinal epithelium. The plasma membrane of these tall columnar cells reflects their high degree of polarization, by dividing into apical and basolateral domains with different compositions. The most characteristic structure of these cells consists of closely packed apical microvilli with the same height, looking like a brush, which is why they were named the brush border. The concentrated pattern of some apical markers observed in a restricted brush border domain shows that mature enterocytes are hyperpolarized epithelial cells: the filamentous brush border glycocalyx is anchored at the top of the microvilli and the annexin XIII is concentrated in the lower three fourths. Many studies have been carried out on the biosynthesis and intracellular pathway of domain markers. The results show clearly that the basolateral markers take a direct pathway from the trans-Golgi network to the basolateral membrane. However, the two apical pathways, one direct and one indirect pathway via the basolateral membrane, are used, depending on the apical protein involved. Efficient protein sorting and addressing are essential to the establishment and maintenance of cell polarity, on which the integrity of the epithelial barrier depends. PMID- 10820520 TI - Developmental aspects of lipid and lipoprotein synthesis and secretion in human gut. AB - This review article focuses on the ontogeny and the regulatory mechanisms involved in the modulation of the intracellular events governing the assembly and delivery of lipoproteins in human gut. The human fetal intestine organizes villi covered with well-differentiated enterocytes during the end of the first trimester in utero. One striking event is the formation of villi in the colonic mucosa similar to those of the small intestine. The small intestine exhibits very early (14-20 weeks) the capacity to absorb lipids, to elaborate most of the major lipoprotein classes (chylomicrons, very-low-density lipoproteins, low-density lipoproteins, high-density lipoproteins), and to efficiently export these lipoproteins from the intestinal cells. The ontogenic changes of lipid and lipoprotein synthesis are correlated with specific patterns of regulatory enzymes (HMG-CoA reductase, ACAT, MGAT) that are representative of key patterns such as the cholesterol pathway, cholesterol esterification, and neutral lipid pathway. The human fetal colon also has the capability to synthesize lipids, lipoproteins, and apolipoproteins. However, comapred with the small intestine, it is much less efficient at exporting these lipoproteins. Epidermal growth factor, insulin, and hydrocortisone, which are known modulators of the brush border digestive functions of the human gut, differentially modulate the synthesis and secretion of lipoproteins in the small intestine and colon. The use of human fetal gut represents a unique model to further our understanding of the complex biosynthetic molecular events essential for the formation and secretion of lipoproteins relevant to human intestine, both in normal or pathological conditions. PMID- 10820521 TI - Use of immunoelectron microscopy and intestinal models to explore the elaboration of apolipoproteins required for intraenterocyte lipid transport. AB - The intestine is the organ that contributes the majority of circulating alimentary lipoproteins. Intestinal epithelial cells have the unique ability to elaborate chylomicrons, the largest triglyceride-rich lipoproteins and the main vehicle for the transport of dietary lipids. The final intracellular assembly and exocytosis of chylomicrons require enterocyte-derived apolipoproteins (apo). As research on lipoprotein metabolism evolved, it has become increasingly evident that apo B is a crucial protein for the normal packaging of triglyceride-rich lipoproteins. Immunocytochemical techniques have successfully been used to demonstrate the presence of two types of apo B, the B-100 and the B-48, in different subcellular compartments of the human enterocyte. Confirmation was obtained by biochemically analyzing human lymph and intestine from pediatric patients. In addition, the immunoelectron microscopic approach revealed the location of apo A-I in the rough endoplasmic reticulum (ER) and predominantly in the Golgi apparatus and the basolateral membrane, which confirms the rapid transport of apo A-I documented by other studies. Proven utility and experimental conditions were defined to demonstrate the ability of Caco-2 cells, a colon carcinoma cell line, to esterify lipids, synthesize apo, and assemble lipoproteins. Thus, immunocytochemical and biochemical techniques can be combined with in vivo and in vitro intestinal models for the study of the intestinal lipid transport. PMID- 10820522 TI - Electron microscopic improvement in the study of diarrheagenic Escherichia coli. AB - In the versatile single species of Escherichia coli, the diarrheagenic group displays a remarkable array of virulence traits. These comprise microbial attachment, production of secretory endotoxins or cell-destroying cytotoxins, direct epithelial cell invasion, and localized effacement of the epithelium. The knowledge of how enteric E. coli induce disease has become increasingly important in the world, because of new pathogen emergence, increasing threats of drug resistance, and growing awareness of their importance in malnutrition and diarrhea. Numerous research programs have demonstrated various mechanisms of pathogenesis. We point out how some pathogens are able to develop intercourse with their host through subversion of its cytoskeleton and signaling processes without toxin secretion or heavy invasiveness. In that domain, the cellular biology of infected cells owes fundamental data to the electron microscopic approach. Combined with advances in microbiology and molecular biology, this approach may provide answers to many unanswered questions. PMID- 10820523 TI - Human cell models to study small intestinal functions: recapitulation of the crypt-villus axis. AB - The intestinal epithelium is continuously and rapidly renewed by a process involving cell generation, migration, and differentiation, from the stem cell population located at the bottom of the crypt to the extrusion of the terminally differentiated cells at the tip of the villus. Because of the lack of normal human intestinal cell models, most of our knowledge about the regulation of human intestinal cell functions has been derived from studies conducted on cell cultures generated from experimental animals and human colon cancers. However, important advances have been achieved over recent years in the generation of normal human intestinal cell models. These models include (a) intestinal cell lines with typical crypt cell proliferative noncommitted characteristics, (b) conditionally immortalized intestinal cell lines that can be induced to differentiate, and (c) primary cultures of differentiated villuslike cells that can be maintained in culture for up to 10 days. Each of these models should help in the investigation of the specific aspects of human intestinal function and regulation. Furthermore, taken together, these models provide an integrated system that allows an in vitro recapitulation of the entire crypt-villus axis of the normal human small intestine. PMID- 10820524 TI - HPV primary screening for cervical cancer: more pain than protection. PMID- 10820525 TI - Diagnosis and classification of lymphoma based on cytospin preparations: a comparison of hematopathologists and cytopathologists. AB - The diagnosis of malignant lymphoma based on cytologic preparations is a source of much debate. The purpose of this study was to assess the ability of a number of pathologists to diagnose and classify lymphoma using cytospin preparations, and to compare the rate of agreement between cytopathologists and hematopathologists. One hundred twenty-five cytospins prepared from histologically confirmed hematologic lesions were examined retrospectively and independently by four hematopathologists/fellows and two cytopathologists without knowledge of the final diagnosis; the results were compared with the final diagnoses derived from histology and immunophenotyping. Eighty-one cases were histologically diagnosed as lymphoma (including 67 cases of B-cell non-Hodgkin's lymphoma), and 44 cases represented a reactive process histologically. The distinction of a malignant from a benign process was made in 75% of the cases by cytospin examination, with cytopathologists correctly diagnosing 75% and hematopathologists 76% of the cases. The accuracy rate for subclassification of the lymphoma cases was 49% (46% for cytopathologists, 52% for hematopathologists). The cytopathologists correctly recognized large-cell lymphoma at an increased frequency compared with the hematopathologists (70% vs. 56%, P = 0.11), while the hematopathologists showed a greater ability to recognize and classify nonfollicle center low-grade B-cell lymphomas (57% vs. 28%, P = 0.01). We conclude that cytopathologists and hematopathologists generally achieve similar accuracy rates in the morphologic evaluation of cytologic preparations of lymphoid lesions, though some differences in their performance do exist. PMID- 10820526 TI - Endocervical reactive atypia: a histologic-cytologic study. AB - Of 1,500 cervical tissue specimens, 27 cases showed histologic changes of reactive glandular atypia which we defined as endocervical cells with large hyperchromatic, often irregular nuclei, which did not fulfill the criteria for endocervical adenocarcinoma. Eighteen of these 27 cases had preceding or concurrent cervico-vaginal smears. Six of these showed cells which were similar to those seen in histologic sections. The cytologic characteristics of these cells are defined. To determine if atypia is related to inflammatory-regenerative changes, 29 cases of endocervical polyps were examined, of which 11 showed histologic changes of endocervical reactive atypia; 4 showed these changes cytologically as well. Twenty-eight cases of routine hysterectomy specimens were examined, of which 2 cases showed endocervical reactive atypia, which indicated that the atypical changes were indeed reactive. Nine out of 27 cases were associated with hormonal usage. Fourteen cases were associated with squamous intraepithelial lesions or evidence of human papilloma virus. Follow-up of our 27 index cases revealed no progression to adenocarcinoma. These findings indicate that atypia, as we define it, of the endocervix can be due to inflammatory reparative changes or possibly related to hormonal usage, and permit its separation from precursor lesions of endocervical adenocarcinoma. PMID- 10820527 TI - Relation between natural killer cells and neoplastic cells in serous fluids. AB - Natural killer (NK) cells are capable of lysing certain tumor cells in vitro. We investigated whether the percentage of NK cells (NK%) in serous effusions has any relation with the presence of malignant cells. Fresh effusions were routinely processed, and a portion of them was submitted for lymphocyte immunophenotyping, using a flow cytometer. Fifty-one fluids (15 peritoneal, 36 pleural), from 47 patients were analyzed. Thirty-two cases were negative and 19 were positive for neoplastic cells: there were 13 adenocarcinomas, 2 malignant mesotheliomas (MM), 3 malignant lymphomas (ML), and 1 rhabdomyosarcoma (R). Negative cases showed an average NK% of 5.09% (1-16%). Effusions positive for adenocarcinoma had an average NK% of 22.08% (12-33%) (P < 0.001). MM, ML, and R had a low NK%, with an average of 3.01% (0-5%). Investigation of the NK% could be useful in cytologically doubtful cases and in the differential diagnosis between MM and adenocarcinoma. PMID- 10820528 TI - Cytologic diagnosis of medullary carcinoma of the thyroid gland. AB - The cytomorphologic features in fine-needle aspiration (FNA) biopsies from 91 histologiacally verified medullary carcinomas of the thyroid (MCT) were investigated. FNA was able to diagnose neoplasms with indications of surgical removal in 98.9% of cases and moreover, was accurate in specific tumor typing in 89% of cases. The most important cytologic criteria of MCT with FNA are: dispersed cell-pattern of polygonal or triangular cells, azurophilic cytoplasmic granules, and extremely eccentrically placed nuclei with coarse granular chromatin and amyloid. These and other cytologic features of MCT are discussed in detail. Fourteen cases of thyroid tumors originally diagnosed as MCT by cytology are illustrated to discuss the differential diagnosis of MCT and its potential pitfalls. If MCT is cytologically presumed but amyloid and azurophilic cytoplasmic granules are not demonstrated, the use of immunostaining is necessary for a correct tumor typing. The application of immunocytochemistry in MCT is discussed. PMID- 10820529 TI - Proliferating ductules are a diagnostic discriminator for intrahepatic cholangiocarcinoma in FNA biopsies. AB - The histologic distinction of cholangiocarcinoma (CC) from metastatic carcinoma and hepatocellular carcinoma (HCC) is difficult. In particular, the distinction of CC from metastatic carcinoma on morphologic features alone is not possible and is dependent on the identification of an extrahepatic primary carcinoma. The proliferative response to many types of liver injury is characterized by a proliferation of either hepatocyte ductular clusters (HDC) or biliary ductular clusters (BDC). This study examined the presence of such ductular reactions in fine needle aspiration biopsies of 20 consecutive cases each of CC and HCC, and compared the findings to those of 20 cases of hepatic metastases from a wide variety of sites. All 18 cases of CC with adequate smears showed ductular proliferation of either HDC or BDC type, the latter being more common; in 13 cases, there were more than 10 ductular clusters per smear. In contrast, only one case of metastatic carcinoma displayed so many ductular clusters, this being a case with multiple hepatic deposits. Five cases of HCC showed more than 10 clusters. The presence of more than 10 ductular clusters associated with malignant cells is a useful discriminator to separate CC from metastatic carcinoma. PMID- 10820530 TI - Malignant phyllodes tumor with liposarcomatous components: a case report with cytological presentation. AB - Cytological and histological findings of a rare case of a malignant phyllodes tumor (with liposarcomatous components) of the breast are presented. The atypia of the stromal cells and naked nuclei in a lesion considered benign clinically and on imaging alerted us to the possibility of a phyllodes tumor, despite the low cellularity of the preoperative fine-needle aspiration smears. The excisional biopsy was histologically diagnosed as malignant phyllodes tumor with liposarcomatous components. Peculiar atypical cells with large, pale, transparent cytoplasm, or with intranuclear chromatolytic areas similar to intranuclear cytoplasmic inclusions, in addition to atypical lipoblasts revealed in imprint smears from the excised tumor, may be important for cytopathologists to achieve a definitive cytological diagnosis, and also to observe patients over long periods for recurrences and metastasis after surgery for this rare breast tumor. PMID- 10820531 TI - Cytomorphologic features of classic and variant lobular carcinoma: A comparative study. AB - There are several subtypes of lobular carcinoma (LC), and their cytomorphologic features differ from classic lobular carcinoma (CLC). The finer details of the differences between CLC and variant lobular carcinoma (VLC) have not been adequately studied. A comparative study of 54 cases of CLC and VLC was done in order to verify any statistically significant differences between them. All cases had histologic confirmation of the diagnosis. Six parameters, which included cellularity, signet-ring cells, intracytoplasmic lumina (ICL), anisonucleosis, cell size, and prominent nucleoli, were studied. The only statistically significant findings were cellularity and cell size when compared to CLC. The cellularity in VLC was higher and the cells in VLC were larger when compared to CLC. There are no definite diagnostic features to identify VLC; however, in a cellular specimen with plenty of large cells with other features of LC, one should have a high index of suspicion of VLC. PMID- 10820532 TI - Aspiration cytology and core biopsy of a carcinoid tumor arising in a retrorectal cyst: a case report. AB - Retrorectal cysts are uncommon lesions of uncertain histogenesis, and primary carcinoid tumors arising in retrorectal cysts are extremely rare. We present the case of a 52-yr-old man who had a 22-cm partially cystic, partially solid mass in the presacral space. A computed tomography-guided fine-needle aspiration of the mass was performed. The smears contained abundant keratinous debris and rare groups of tumor cells. The tumor cells were cuboidal, with slightly granular cytoplasm and centrally located nuclei with speckled chromatin and inconspicuous nucleoli. Immunocytochemical analysis revealed strong reactivity for chromogranin and keratin, and focal reactivity for synaptophysin and neuron-specific enolase. The cytological diagnosis of a carcinoid arising in a tail-gut cyst was confirmed histologically. This is the first reported case of a carcinoid arising in a retrorectal cyst diagnosed preoperatively by cytology. This diagnosis is possible in the setting of consistent clinical, radiographic, and cytological findings. PMID- 10820533 TI - Granular-cell tumor of trachea masquerading as Hurthle-cell neoplasm on fine needle aspirate: a case report. AB - We report on a case of extraluminal tracheal granular-cell tumor which was interpreted as a Hurthle-cell neoplasm of the thyroid on fine-needle aspirate. Review of the literature reveals only one other such case. The patient was a 35 yr-old female who presented with an enlarged thyroid. Aspiration cytology revealed a syncytium of cells with abundant granular cytoplasm interpreted as a thyroid follicular neoplasm with Hurthle-cell change. However, histology of the resection specimen with immunohistochemistry confirmed it as a granular-cell tumor. The cytologic differential diagnosis of neoplasms with oncocytoid cytoplasm in and around the thyroid should include granular-cell tumor of the trachea. PMID- 10820534 TI - Myxoid chondrosarcoma of the sphenoid sinus and chondromyxoid fibroma of the iliac bone: cytomorphologic findings of two distinct and uncommon myxoid lesions. AB - Myxoid chondrosarcoma (MCS) and chondromyxoid fibroma (CMF) are two uncommon myxoid cartilaginous neoplasms with distinct cytologic features, histologic patterns, and immunoprofiles. Because these neoplasms have characteristic biological behaviors and management, their correct diagnosis is crucial to avoid debilitating and unnecessary surgical procedures. We report the imprint cytology (IC) preparation findings along with the differential diagnosis in one case each of myxoid chondrosarcoma and chondromyxoid fibroma of the splenoid sinus and iliac bone, respectively. The two great mimickers for these neoplasms, chordoma and chondrosarcoma, represent difficult diagnostic challenges, especially when MCS and CMF occur in unusual locations. IC in conjunction with the clinical and radiologic findings can provide a rapid preliminary intraoperative diagnostic interpretation which can aid in planning the immediate surgical management, as well as guide specific tissue triage for key ancillary studies such as electron microscopy and cytogenetic analyses. To the best of our knowledge, there have been no cytologic reports of MCS of the sphenoid sinus and CMF of the iliac bone. PMID- 10820535 TI - Nuclear pseudoinclusions in fine-needle aspiration cytology of hepatic angiomyolipoma: case report. AB - A nodular lesion was found in the fourth segment of the liver in a 51-yr-old Caucasian woman and subjected to aspiration cytology. Smears exhibited solid sheets or small aggregates of epithelioid cells with numerous nuclear pseudoinclusions, scanty mature adipocytes, and red blood cells. With a presumptive diagnosis of angiomyolipoma, the patient underwent laparotomy with resection of the nodule; histologic and immunohistochemical findings confirmed the diagnosis, identifying the trabecular variant. Although the presence of nuclear pseudoinclusions has been previously reported in renal angiomyolipomas, this cytologic feature has not received adequate attention in liver localization, where only cell block samples have been reported. Therefore, when the typical different cellular components of angiomyolipomas are absent in cytologic smears, nuclear pseudoinclusions should not be regarded as a criterion of malignancy, but they may be considered as an additional nonspecific cytological feature in hepatic angiomyolipoma. PMID- 10820536 TI - Melanocytoma of the ciliary body diagnosed by fine-needle aspiration biopsy. AB - A 55-yr-old African American man was referred for evaluation of a mass involving the anterior choroid/ciliary body in his left eye. Dilated fundus examination revealed a large, deeply pigmented mass associated with an exudative retinal detachment. Ultrasonography demonstrated an elevated mass which involved the anterior uvea and showed low reflectivity by standardized A-scan. Cytologic examination of fine needle aspiration (FNA) biopsy from the tumor was interpreted as a melanocytoma. The patient developed a total retinal detachment and secondary glaucoma which led to enucleation of the eye. Histopathologic examination confirmed the diagnosis of melanocytoma. To our knowledge, this is the first report in which the diagnosis of uveal melanocytoma was established by FNA biopsy and later confirmed histopathologically. PMID- 10820537 TI - Follicular carcinoma of the thyroid with extensive clear-cell differentiation: a potential diagnostic pitfall. AB - Clear-cell features have been recognized in several different thyroid neoplasms. A case of thyroid follicular carcinoma with extensive clear- and Hurthle-cell features is described in a 37-yr-old white female, with cytochemical and immunohistochemical analysis. The tumor of the thyroid gland, with anterior neck soft-tissue extension, displayed clear cells on fine-needle aspiration, which were negative for thyroglobulin. The surgical specimen displayed predominately clear cells (80%), and only the nonclear-cell areas stained for thyroblobulin. Proper categorization of clear-cell lesions of the thyroid and soft tissues requires a multimodality approach, involving clinical/pathological correlation, morphological analysis, and ancillary tissue studies. Immunohistochemical stains for thyroglobulin are quite definitive in making the distinction between primary clear-cell thyroid tumors vs. metastatic clear-cell tumors. Cytologists should be aware, however, that the clear-cell areas of thyroid tumors might not stain for thyroglobulin. PMID- 10820538 TI - Lymphoepithelial carcinoma of salivary gland - cytologic, histologic, immunocytochemical, and in situ hybridization features in a case. PMID- 10820539 TI - Electronic publication of biomedical research. PMID- 10820540 TI - Cybermedicine: promises and perils. PMID- 10820541 TI - T-lymphocyte subpopulations in tuberculosis. AB - BACKGROUND: Tuberculosis is associated with both qualitative and quantitative defects in the cell mediated immune response. The changes that occur in the lymphocyte profile in blood in children with tuberculosis are not well understood. DESIGN: Prospective study. SETTING: Referral hospitals. METHODS: Lymphocyte subpopulations were determined by flow cytometry in 17 healthy tuberculin positive children, in 22 children with newly diagnosed pulmonary tuberculosis and in 8 of these children after antituberculosis therapy. RESULTS: Absolute numbers and percentages of CD3+ and CD4+ T cells were reduced in children with tuberculosis, compared to controls. CD4+ counts increased significantly following antituberculosis treatment, compared to baseline values. In contrast, the proportion of T cells expressing the gdT cell receptor was similar in tuberculosis patients and controls. CONCLUSION: Children with tuberculosis have a systemic decrease in the proportion and number of CD3+ and CD4+ T cells which reverses during therapy. PMID- 10820542 TI - Bone density values in healthy Turkish infants. AB - OBJECTIVE: To determine values of bone density of Turkish infants under one year of age. DESIGN: Cross sectional. METHODS: This study included 1, 2, 4, 6, 9 and 12 months old 164 male and female healthy infants who were evaluated by dual energy X-ray absorbtiometry. RESULTS: The values for each age were computed. There was no significant difference between boys and girls. A significant relationship was found between bone mineral density, bone mineral content, total body calcium and calcium/body surface, and age, height, body weight, head circumference and body surface area. Multiple regression analysis showed that body mineral density was affected by weight, height, head circumference and body mass index. Body mineral content and total calcium content were affected by weight and height. CONCLUSION: The results will serve as reference values for healthy 1-12 months old Turkish infants. PMID- 10820543 TI - Scorpion sting. PMID- 10820544 TI - Measures of mortality and morbidity in children. PMID- 10820545 TI - Use of high frequency ventilation as a rescue measure in premature babies with severe respiratory failure. PMID- 10820546 TI - Hemophagocytic lymphohistiocytosis: a case series. PMID- 10820547 TI - Prevalence of anemia among adolescent girls of rural Tamilnadu. PMID- 10820548 TI - Iodine deficiency disorders in Bargarh district of Western Orissa. PMID- 10820549 TI - ANA negative lupus erythematosus. PMID- 10820550 TI - Aicardi's syndrome in a male child: an unusual presentation. PMID- 10820551 TI - Myoclonic epilepsy with ragged red fibres. PMID- 10820552 TI - Candidial intestinal perforation in a non-immunocompromised infant. PMID- 10820553 TI - Septic arthritis due to Ureaplasma urealyticum. PMID- 10820554 TI - Subcutaneous nodules. PMID- 10820555 TI - Dose of varicella vaccine. PMID- 10820556 TI - Dose of varicella vaccine - reply PMID- 10820558 TI - HIV infection and immunization - reply PMID- 10820557 TI - HIV infection and immunization. PMID- 10820560 TI - HIV infection prevention in specific situations PMID- 10820559 TI - HIV infection prevention in specific situations. PMID- 10820562 TI - BETARU: an indigenously designed cordless stethoscope. PMID- 10820561 TI - Neonatal pericardial tamponade from a silastic central venous catheter. PMID- 10820563 TI - Management of beta thalassemia. PMID- 10820564 TI - Management of beta thalassemia - reply PMID- 10820565 TI - Intramuscular vaccination in hemophilia. PMID- 10820566 TI - Intramuscular vaccination in hemophilia - reply PMID- 10820567 TI - Clinical profile and sero-conversion pattern of children with HBsAg positivity. PMID- 10820568 TI - Clinical profile and sero-conversion pattern of children with HBsAg positivity PMID- 10820569 TI - The attitude of nurses towards breastfeeding--a cross-sectional survey. PMID- 10820570 TI - Anti-cholinergic drugs and wheezy infants. PMID- 10820571 TI - External examining: the challenge for clinically-based degree programmes. PMID- 10820572 TI - An exploration into the role of the teacher/lecturer in practice: findings from a case study in adult nursing. AB - Against a background of increasing concern over the widening theory practice gap and research evidence of problems in providing quality practice education, the English National Board (ENB) commissioned an 18 month study into the role of the teacher/lecturer in practice. This article, after providing a brief overview of the background, literature and methods, outlines the results from the Adult Case Study stage of the research. Documents, individual and focus group interviews of lecturers (n = 76), practitioners (n = 46) and students (n = 131) in five schools of nursing and midwifery provided data for analysis. A role labeled as 'link lecturer' was the most common approach acknowledged by all respondent groups. Despite overt commitment to the practice role by schools of nursing and midwifery, findings indicate that lecturers are unprepared, unsupported and unmonitored. Students and practitioners had very variable experiences and expressed a strong need for a better-organized approach which would provide them with consistent and sustained support in the practice setting. Overall the study highlighted a lack of strategic management of the practice role by university schools of nursing and midwifery. Convergence of findings from midwifery and the other branches of nursing provides strong research evidence for recommendations for more active management of the practice curriculum to be implemented as part of any future proposed reforms for nurse education in the UK. PMID- 10820573 TI - Articulation of pre-registration nursing courses in Western Sydney. AB - This paper will describe a project, undertaken in 1996-1997, which sought to develop formal articulation arrangements between nursing education courses provided by Technical and Further Education (TAFE) institutions in Greater Western Sydney and the University of Western Sydney (UWS). The project sought to facilitate the access of TAFE-trained Enrolled Nurses (ENs) to UWS's Bachelor of Nursing (BN) courses and eliminate costly duplication in programs of study. Although conducted in Australia, the project generated insights of utility to colleagues in the UK, the USA and New Zealand, in particular, who are dealing with the effects of the 'knowledge explosion' and reductions in government funding to universities. The project comprised two stages. Stage 1 involved a statewide survey of all stakeholders, using individual telephone interviews and focus groups, of practices and perceptions with respect to formal articulation arrangements. The findings demonstrated an overwhelming consensus with respect to both the benefits of and barriers to formal articulation arrangements. These benefits and barriers will be described. Stage 2 involved the TAFE-UWS collaborative design and implementation of 2 EN-BN bridging programs which allow, differentially, ENs to access UWS's BN programs with 1 and 2 semesters block credit. This paper includes details of the project's background, survey methods and findings, bridging courses' contents and courses' implementation evaluative data. PMID- 10820574 TI - Disability as an equal opportunity issue within nurse education in the UK. AB - Whilst equal opportunity policies, related research and subsequent curricula changes over recent years may have begun to have a positive impact in some areas of nursing education and practice, 'disability' struggles for recognition as an equality issue. Disabled people, making up approximately six million of the UK population, experience discrimination as part of their daily lives and this is also evident in the nursing services they receive. Far from challenging discrimination, nursing education may simply sustain the negative dominant ideological view of disability. Drawing comparisons with 'race', which has an established equal opportunity dimension, this paper explores disability equality, which is relatively neglected in nursing curricula. From this review it is argued that nursing as a profession has a role beyond patient care in influencing public perceptions concerning disabled people and should take more positive steps in promoting social justice by challenging discrimination. Stakeholders in nursing curricula and nursing practice have both legal and professional responsibilities to ensure that nursing and nurses become part of the solution to disability discrimination. Including disability in equal opportunity policies and strengthening their impact on pre and post registration nursing curricula, provides nursing with a tangible opportunity to do just that. PMID- 10820575 TI - Implementing evidence-based health care: the nurse teachers' role in supporting the dissemination and implementation of SIGN clinical guidelines. AB - The effectiveness of clinical care and treatment is central to the quality of health care. Clinical guidelines, audit and research are interrelated activities central to the delivery of high quality services and care. A culture is required that supports the implementation of clinical guidelines and encourages clinicians to critically appraise and evaluate their practice and to become more involved in clinical audit and research. An essential element in the effective implementation of clinical guidelines is educational support. This paper will describe the results of a study which explored the nature of the Scottish Intercollegiate Guidelines Network (SIGN) and the nurse teachers' role in supporting the dissemination and implementation of clinical guidelines within acute care nursing and subsequent curriculum design. PMID- 10820576 TI - An evaluative study of clinical preceptorship. AB - Clinical preceptorships, in collaboration between clinical agencies and educational institutions have been documented as an effective and innovative means of facilitating student learning, providing advantages for both the clinical and educational settings. A preceptorship programme of 100 hours duration was developed and delivered by the nurse education institute, in consultation with a health care organization. The objectives of the preceptorship programme were to help registered nurses, in partnership with clinical nurse educators, to effectively integrate, support and assist the development of clinical competence in the undergraduate nursing student. Following the implementation of the preceptorship programme a research study was conducted to evaluate programme effectiveness. The purpose of the study was to examine preceptors, preceptees, and nurse managers' preceptions of the preceptor role and factors which influenced the performance of preceptors. The methods used in this study included those commonly found in evaluation research. That is, participants were drawn from those who were involved, either directly or indirectly, in the preceptorship programme, namely preceptors, preceptees and nurse managers. Using focus groups, they were each asked to identify the outcomes of the programme in practice. Study findings highlighted the importance of formal preceptor preparation, which was shown to enhance teaching and learning opportunities for student preceptees, personal and professional development of the preceptors, and the promotion of positive partnerships between nurse educators and nurse practitioners. The need for formal recognition of the preceptor role in practice, particularly in relation to the provision of adequate time and resources, emerged from the study. The research findings enabled the development of an evaluative model of preceptorship, which highlights the intrinsic and extrinsic factors impacting on the preceptor role. PMID- 10820577 TI - 'We don't sing the chorus when the folksinger's here': the learning society and health care. AB - This paper reports on a study carried out as part of the Economic and Social Research Council's (ESRC's) Learning Society research programme. The study focused on continuing education and training (CET) in Scotland and covered four sectors, of which one was health care. The paper reports on qualitative evidence gathered from focus groups and individual interviews. It deals with three key issues: initial education and the implications for continuing education and training; markets, investment and the returns to CET; and cultural factors. The paper concludes with some key implications. PMID- 10820578 TI - The way forward for transcultural nursing. AB - This paper suggests that the incorporation of a sociological perspective into transcultural nursing would enhance the present anthropological approach. The discussion considers the concept of ethnicity, which encompasses a wider perspective than that of culture alone, and is inclusive of both dominant and subordinate human social groupings. Anthropological understanding explores the beliefs and values of a particular community whereas sociological understanding reflects the position of that community within society and their associated experiences. The case is made by discussion of transcultural nursing, its roots in anthropology and the contribution that a sociological perspective can provide. Concepts of assimilation and acculturation are discussed, as is the adaptive nature of culture. A need for application of theoretical understanding to professional practice is demonstrated and a requirement for development of cultural competence is highlighted. The implications for curriculum development are discussed. This paper adopts a perspective that considers transcultural health care from the focal point of the user of health care service rather than the more usual dominant provider viewpoint. PMID- 10820579 TI - Education about pain: a neglected area? AB - To test the hypothesis that educational deficits are in part responsible for the low priority given to pain management by nurses, a study was carried out to ascertain the pain content in the Common Foundation Programme (CFP) and the four branches of pre-registration diploma nursing courses in England. Questionnaires were sent to branch co-ordinators at all the institutions (n = 47) in England which provide the three year pre-registration nursing diploma courses. Results indicate that while child and adult branch programmes cover a wide breadth of topics in on average less than 10 hours. Learning disabilities branch and mental health branch programmes have minimal pain content. These results suggest that there is superficial coverage of the topic. There is need to re-evaluate the teaching of pain in pre-registration nursing courses if pain and its management are to be adequately covered prior to qualification. These results may also have implications for patient welfare and are discussed in this context. PMID- 10820580 TI - Interaction of monensin and sulfadimethoxine in broilers, as mediated by hepatic microsomal cytochrome P-450 monooxygenases. AB - The influence of monensin + sulfadimethoxine on cytochrome P-450 monooxygenase activity in broilers, and the possible consequences of modification of this system, including changes in blood levels of sulfadimethoxine, influence on the duration of xylazine-ketamine anesthesia, total antioxidant status and superoxide dismutase activity were studied. The results indicate that the combination of monensin + sulfadimethoxine gave a short-term inhibition of microsomal cytochrome P-450 monooxygenase activity but apparently did not influence the metabolism of other (exogenic) substances (ketamine, xylazine), and did not change the state of antioxidant systems or the relative liver weight. There was a rise in blood sulfadimethoxine levels. PMID- 10820581 TI - The relevance of ethnic influences on pharmacogenetics to the treatment of psychosis. AB - Interethnic variation amongst the drug metabolising enzymes relevant to the treatment of psychosis is reviewed. The frequency of genetically determined variants at the extremes of enzyme activity is seen to vary considerably between different ethnic groups; in addition, a shift in the frequency distribution giving an overall lower population mean activity may occur. The role of dietary and other environmental influences in the generation of interethnic variation in cytochrome activity is also discussed. Clinical studies pertinent to this variation are reviewed. It is suggested that the reason for conflicting data from some clinical studies is the existence of overlapping substrate specificity, so that one cytochrome is able to substitute for another. Individuals deficient for more than one cytochrome would be likely to show much more pronounced clinical effects than those showing single cytochrome deficiency. PMID- 10820582 TI - The influence of ethnicity and antidepressant pharmacogenetics in the treatment of depression. AB - Antidepressant disposition can be influenced by a variety of CYP isozymes and their effects in the treatment of depression are reviewed. The CYP isozymes 2D6, 3A4, 1A2 and 2C are discussed in regard to antidepressant drug pharmacokinetics, clinical relevance and variability in activity for each isozyme. Polymorphism has been identified with CYP 2D6 and 2C19. Disposition of antidepressants which are substrates of these two isozymes can also be influenced and contributes towards the wide interpatient and interethnic variability found with these drugs. Antidepressants (especially SSRIs) can be CYP isozyme inhibitors and produce significant drug-drug interactions. PMID- 10820583 TI - Psychopharmacology research in the English-speaking Caribbean. AB - Ethnic differences in patients' responses to drugs used to treat psychiatric disorders have been increasingly reported. Differences can be found in the English-speaking Caribbean population due to its multi-ethnic composition. With the goal of identifying studies on psychopharmacology and ethnicity, we searched for all abstracts and full papers on clinical psychopharmacology emerging from research in the English-speaking Caribbean published in the West Indies Medical Journal, the Caribbean Medical Journal and Medline. We found no studies focusing on this topic, although there were some which may indirectly imply the existence of such differences. High blood levels of imipramine have been found in Barbadian patients, and the majority of cases of neuroleptic malignant syndrome have been reported in Africans, as well as high levels of CPK. However, the numbers of patients studied have not been large enough to be representative. A significantly higher incidence of alcoholism has also been reported in males of East Indian origin. We concluded there is an urgent need in the region for undertaking this type of research to ensure a more favorable treatment outcome of patients treated with such drugs. PMID- 10820584 TI - Modality effects and the development of the word length effect in children. AB - Two experiments investigated the development of the word length effect in children aged 4 to 10 years, comparing auditory and visual stimuli. The question addressed was whether word length effects emerged earlier with auditory presentation or visual presentation, or whether they emerged at the same age regardless of presentation modality. Results provided evidence that word length effects emerge earlier with visual than auditory presentation. The implication of our results is that with visual presentation, 4-year-olds engage in some form of verbalization strategy that involves obtaining phonological representations of picture names and mapping them on to articulatory output plans. This strategy is clearly verbal in nature, but is not necessarily characterised as cumulative verbal rehearsal. PMID- 10820585 TI - A developmental trajectory in implicit memory is revealed by picture fragment completion. AB - Dissociations between performance on implicit and explicit tasks have often been taken as evidence that different neural structures subserve the two types of memory. One such dissociation involves developmental differences that emerge in explicit tasks, but which appear to be absent in implicit tasks. Such findings are consistent with the idea that implicit memory is subserved by a more primitive system that evolves earlier at both phylogenetic and ontogenetic levels. The present paper reviews previous studies that claimed to find evidence that implicit memory is fully developed in very young children. Issues of measurement error, ceiling effects, and insufficient power brought up questions about those studies with respect to the developmental issue. The present study compares performance on implicit (picture fragment completion) and explicit (free recall and recognition) memory tasks with groups ranging in age from 5-28 years. We find a reliable developmental trend in both implicit and explicit performance in which the former cannot be attributed to the operation of explicit memory processes. Thus, we conclude that implicit memory, like explicit memory, develops with age. PMID- 10820586 TI - Remembering the street names of one's childhood neighbourhood: a study of very long-term retention. AB - Life-span retention of street names was studied in a sample of former students of a Dutch elementary school. Participants were requested to recall the street names of their childhood neighbourhood and indicate their position on a map. In addition, information was gathered concerning (a) the extensiveness of the original learning experience, (b) its elaborateness, and (c) the amount of interference from similar materials occurring between original learning experience and time of recall. Retention intervals varied from 0 to 71 years. Amount of exposure, elaborateness of learning, and retroactive interference all contributed to the memorability of names. In addition, the forgetting curve showed a permastore effect (Bahrick, 1984), suggesting that memory for non schematic, incidentally learned material is subject to processes of forgetting similar to those that affect intentionally learned material, such as subject matter acquired in school. PMID- 10820587 TI - Working memory capacity and time course of predictive inferences. AB - A naming task assessed activation of inference concepts during reading. A predicting, or a control, context sentence was followed by a target word to be named, which represented the predicted event or an inconsistent event. The interval between the end of the context and the onset of the target word varied between 50 and 1050 msec. Individual differences in working memory capacity were assessed by the reading span task. As reflected by facilitation in naming latencies in the predicting condition, relative to the control condition, (a) inferences were not made within the first 50 msec after the context, regardless of reading span; (b) only the high-span participants drew inferences within a 550 msec interval; and, (c) both the high- and the low-span participants generated them within a 1050-msec interval. These results indicate that high working memory capacity accelerates the time course of predictive inferences, although they do not become automatic. We propose that this effect occurs because these inferences involve time-consuming elaborations that place demands on the effortful and limited resources of working memory. Deficiencies in word knowledge, speed of lexical access, or comprehension of explicit information do not account for low span readers' difficulties in generating predictive inferences. PMID- 10820588 TI - A code-theoretical note on object handedness. AB - This study is a theoretical exercise dealing with discrimination between images and mirror-images. It focuses on the way codes of shapes represent their handedness. We compare two code systems with different reference frames. These frames determine the specific sensitivity of each system. One system uses an asymmetric reference frame. It is called the H-system and was inspired by an idea of Corballis (1988 Psychological Review 95 115-123). The other system, being our proposal, uses a symmetric reference frame and we have named it the M-system. We demonstrate the following. A code of the H-system provides a cue for the handedness of a shape, but not for rotation, i.e. no cue for the appropriate kind of code rotation which should be tested in case images and mirror-images are discerned by mental rotation. The M-system is the converse in both respects. A code of this system does not provide a handedness cue but, instead, a rotation cue. Thus, for handedness discrimination, the H-system neither needs nor guides mental rotation, whereas the M-system does both. This M-system generates object centred structural codes enriched with viewpoint information. Various visual experiments reported in the literature favour the M-system over the H-system, implying that perception does not make use of an asymmetric but of a symmetric reference frame. PMID- 10820589 TI - Detection of bilateral symmetry in complex biological images. AB - The recognition of bilateral symmetry in simple dot patterns is reliably influenced by orientation. Performance is best when the axis of symmetry is vertical. We conducted two experiments to determine whether stimulus orientation also affects detection of the low levels of naturally occurring asymmetry in complex biological images. University students judged whether colour images displayed on a computer monitor possessed perfect bilateral symmetry. Stimuli were generated from high-resolution plan-view images of crabs and insects. In experiment 1, the asymmetric stimuli were the original animals, displayed on a standard black background. Symmetrical versions of each natural image were generated by sectioning the shape at the midline, copying and reflecting one side and then fusing the two halves together. To facilitate comparison of results with those obtained in earlier studies, we also presented dot patterns based upon both the slightly asymmetric and perfectly symmetrical natural images. Experiment 2 was designed to assess whether symmetry detection was dependent upon the markings and patterns on the body and appendages of the animals. The natural images were converted to silhouettes and tested against matched dot patterns. In both studies, images were presented in a random sequence with the axis of symmetry vertical, horizontal, oblique left, and oblique right. Performance with the biological images was consistently better than with the dot patterns. Abolishing fine detail did not appreciably reduce this effect. A pronounced vertical advantage was apparent with all stimuli, demonstrating that this phenomenon is robust despite considerable variation in image complexity. The implications of orientation effects for perception of natural structures are discussed. PMID- 10820590 TI - Route navigating without place recognition: what is recognised in recognition triggered responses? AB - The use of landmark information in a route-navigation task has been investigated in a virtual environment. After learning a route, subjects were released at intermediate points along the route and asked to indicate the next movement direction required to continue the route. At each decision point, three landmarks were present, one of which was viewed centrally and two which appeared in the periphery of the visual field when approaching the decision point. In the test phase, landmarks could be replaced either within or across places. If all landmarks combined into a new place had been associated with the same movement direction during training, subjects performed as in the control condition. This indicates that they did not need to recognise places as configurations of landmarks. If, however, landmarks that had been associated with conflicting movement directions during training were combined, subjects' performance was reduced. We conclude that local views and objects are recognised individually and that the associated directions are combined in a voting scheme. No evidence was found for a recognition of places as panoramic views or configurations of objects. PMID- 10820591 TI - Global-perspective jitter improves vection in central vision. AB - Previous vection research has tended to minimise visual-vestibular conflict by using optic-flow patterns which simulate self-motions of constant velocity. Here, experiments are reported on the effect of adding 'global-perspective jitter' to these displays--simulating forward motion of the observer on a platform oscillating in horizontal and/or vertical dimensions. Unlike non-jittering displays, jittering displays produced a situation of sustained visual-vestibular conflict. Contrary to the prevailing notion that visual-vestibular conflict impairs vection, jittering optic flow was found to produce shorter vection onsets and longer vection durations than non-jittering optic flow for all of jitter magnitudes and temporal frequencies examined. On the basis of these findings, it would appear that purely radial patterns of optic flow are not the optimal inducing stimuli for vection. Rather, flow patterns which contain both regular and random-oscillating components appear to produce the most compelling subjective experiences of self-motion. PMID- 10820592 TI - Direct measurement of the curvature of visual space. AB - We consider the horizontal plane at eye height, that is all objects seen at the horizon. Although this plane visually degenerates into a line in the visual field, the 'depth' dimension nevertheless gives it a two-dimensional structure. We address the problem of intrinsic curvature of this plane. The classical geometric method is based on Gauss's original definition: The angular excess in a triangle equals the integral curvature over the area of the triangle. Angles were directly measured by a novel method of exocentric pointing. Experiments were performed outside, in the natural environment, under natural viewing conditions. The observers were instructed not to move from a set location and to maintain eye height, but were otherwise free to perform eye, head, and body movements. We measured the angular excess for equilateral triangles with sides of 2-20 m, the vantage position at the barycenter. We found angular excesses and deficits of up to 30 degrees. From these data we constructed the metric. The curvature changes from elliptic in near space to hyperbolic in far space. At very large distances the plane becomes parabolic. PMID- 10820593 TI - Comparing contrast-modulated and luminance-modulated masking: effects of spatial frequency and phase. AB - The masking of a sinusoidal test grating by contrast-modulated (CM) gratings could, in principle, be attributable to the presence of a distortion product, injected into the stimulus during some nonlinear transformation at an early level of visual processing (e.g. Nachmias, 1989 Vision Research 29 137-142). If so, CM gratings and luminance-modulated (LM) gratings of similar effective contrast and spatial frequency should mask the detection of sinusoids in a similar fashion. We compared the effects of masking by 1 cycle deg-1 CM gratings [both simple beats (8 + 9 cycles deg-1) and amplitude-modulated gratings (8 + 9 + 10 cycles deg-1)], with those of masking by 1 cycle deg-1 LM gratings of low contrast. We found that: (i) CM and low-contrast LM grating masks yielded similar spatial-frequency tuning functions around the modulation frequency of 1 cycle deg-1; (ii) low contrast LM gratings masked the detection of test sinusoids in a highly phase dependent fashion, while masking by CM gratings did not vary systematically with relative spatial phase. The results suggest that masking produced by CM gratings cannot simply be explained by the presence of a distortion product at the beat or modulation frequency. PMID- 10820594 TI - Performance of early-blind and sighted children on olfactory tasks. AB - The goal of the study was to test whether children with congenital or early-onset blindness outperform sighted children on olfactory tasks. Measures of olfactory sensitivity, odour recognition, and odour labeling were obtained. The results show that the blind children were more proficient at correctly labeling 25 common odours than were matched sighted children. However, the blind were not more sensitive to a target odour, nor more proficient at choosing a correct odour label from a list of four. Together, the data point to a circumscribed advantage of blind children at self-generating and retrieving odour labels and, as such, to a limited, but still compensatory, cognitive function. PMID- 10820595 TI - The discrimination of dog odours by humans. AB - The domestic dog is able to identify both individual conspecifics, i.e. other dogs, and individual interspecifics, e.g. humans, by smell. Whilst humans can recognise individual members of their own species using olfactory information, it is unknown whether they can identify members of any other species in this manner. We examined the ability of humans to identify individual dogs by smell. Twenty six dog owners were required to smell two blankets, one impregnated with the odour of their own dog, the other impregnated with the odour of an unfamiliar dog. Participants were required to indicate which of the odours smelt the strongest, which smelt the most pleasant, and which of the odours belonged to their own dog. Most of the participants (88.5%) were able to recognise the odour of their own dog. They showed no significant bias, however, in responding which of the odours they thought smelt the strongest or most pleasant. The results indicate that dogs produce odours that are individually distinctive to their owners, and highlight the fact that humans can recognise members of another species using olfactory cues--an ability presumably acquired without conscious effort. PMID- 10820596 TI - Trieste in the mirror. PMID- 10820597 TI - The perception and recognition of natural object shape from deforming and static shadows. AB - In this study of the informativeness of shadows for the perception of object shape, observers viewed shadows cast by a set of natural solid objects and were required to discriminate between them. In some conditions the objects underwent rotation in depth while in other conditions they remained stationary, thus producing both deforming and static shadows. The orientation of the light source casting the shadows was also varied, leading to further alterations in the shape of the shadows. When deformations in the shadow boundary were present, the observers were able to reliably recognize and discriminate between the objects, invariant over the shadow distortions produced by movements of the light source. The recognition performance for the static shadows depended critically upon the content of the specific views that were shown. These results support the idea that there are invariant features of shadow boundaries that permit the recognition of shape (cf Koenderink, 1984 Perception 13 321-330). PMID- 10820598 TI - Bottom-up clues in target finding: why a Dalmatian may be mistaken for an elephant. AB - We provide informal psychophysical support for a strategy where bottom-up features guide attention toward a target, and the top-down path interprets hypothetical shapes at the target location--as opposed to a dominant top-down approach. In our survey, for which we used the familiar picture of a Dalmatian dog against a dappled background, (i) 75% of subjects initially found a bulging body which overlaps that of the dog, but final 'top-down' percepts were unexpected: nearly all subjects assigned an incorrect head and limbs to the body; (ii) after random rotation of texture elements overlapping computed features only 45% of subjects reported a bulging body, with a few adding limbs etc. The picture of the Dalmatian dog must therefore contain many bottom-up features--a top-down strategy may find 'incorrect' targets at correct target locations. Computational support for these claims is more easily constructed than one may expect. We could compute at least two bottom-up features, both useful in 3-D surface interpolation from 2-D scenes, which yielded significant values at the location of the Dalmatian dog: anisotropic texture compression and affine texture distortion cues. We therefore conclude that the role of top-down processing is overstated in a traditional example such as the Dalmatian dog picture. PMID- 10820599 TI - The face-inversion effect as a deficit in the encoding of configural information: direct evidence. AB - We report four experiments leading to conclusions that: (i) the face-inversion effect is mainly due to the deficits in processing of configural information from inverted faces; and (ii) this effect occurs primarily at the encoding stage of face processing, rather than at the storage stage. In experiment 1, participants discriminated upright faces differing primarily in configuration with 81% accuracy. Participants viewing the same faces presented upside down scored only 55%. In experiment 2, the corresponding discrimination rates for faces differing mainly in featural information were 91% (upright) and 90% (inverted). In experiments 3 and 4, the same faces were used in a memory paradigm. In experiment 3, a delayed matching-to-sample task was used, in which upright-face pairs differed either in configuration or features. Recognition rates were comparable to those for the corresponding upright faces in the discrimination tasks in experiments 1 and 2. However, there was no effect of delay (1 s, 5 s, or 10 s). In experiment 4, we repeated experiment 3, this time with inverted faces. Results were comparable to those of inverted conditions in experiments 1 and 2, and again there was no effect of delay. Together these results suggest that an 'encoding bottleneck' for configural information may be responsible for the face-inversion effect in particular, and memory for faces in general. PMID- 10820600 TI - Perceptual assumptions and projective distortions in a three-dimensional shape illusion. AB - We examine a shape illusion, in which the balconies of a building appear to tilt up or down, depending on the viewpoint. The balconies are actually level parallelogram shapes, but appear as tilted rectangles. We measured the illusory tilts observed when parallelogram shapes are viewed above the line of sight, using three-dimensional stimuli consisting of parallelograms of various tilts viewed at different orientations. Under perspective projection, parallelism and orthogonality are not preserved. However, perspective distortions alone cannot account for the perceived tilts measured in these experiments, since observers perceived illusory tilts even for stimuli in the frontoparallel plane. We introduce a model, based on the theory that observers assume ambiguously projected three-dimensional angles to be equal to 90 degrees, but revise their predictions on the basis of observation. In the model, perceived tilt is predicted as a weighted sum of the tilts predicted by the assumptions that the shape is rectangular, and that the shape is level (i.e. that the angle between the shape and the vertical backboard is equal to 90 degrees). We prove that it is mathematically impossible for a planar rectangle to share a projection with a nonrectangular parallelogram. A less restrictive assumption that just the two leading internal angles are equal to 90 degrees is suggested as an alternative, and it is further proven that this new configuration of angles leads to a unique perceived tilt. The relative weights in the model reflect the amount that each prediction is revised, and are shown to vary systematically with stimulus orientation. For some observers a better fit was found by replacing the level tilt assumptions with an assumption that physical tilt was equal to the projected tilt. PMID- 10820601 TI - Amplitude and phase in the Muller-Lyer illusion. AB - It has previously been claimed that the Muller-Lyer illusion is the result of low pass spatial filtering. One way to understand this would be that the distribution of amplitudes is what generates this illusion. This possibility was investigated by computing the 2-D Fourier transforms of the two Muller-Lyer stimuli and extracting their phase and amplitude spectra. These spectra were combined to create hybrid spectra having the phase of one Muller-Lyer figure and the amplitudes of the other. Images were then created by computing the inverse Fourier transform of the hybrid spectra. Except in cases where the analysis was performed patchwise on very small patches, the figures generated with the phase spectrum of the stimuli having outward-pointing fins appear the longer. This was also the case when stimuli were generated with flat amplitude spectra. Because they show that the Muller-Lyer illusion does not depend on any particular distribution of amplitudes, these demonstrations do not support the theory that the Muller-Lyer illusion is the result of low-frequency filtering. PMID- 10820602 TI - Depth capture by kinetic depth and by stereopsis. AB - The perceived depth of regions within a stereogram lacking explicit disparity information can be captured by the surface structure of regions where disparity is explicit: stereo capture. In two experiments, observers estimated surface curvature/depth of an untextured object (a 'ribbon') superimposed on a cylinder textured with dots, the cylinder curvature being defined by disparity (stereo depth) or by motion parallax (kinetic depth: KD). With the stereo-defined cylinder, depth capture was obtained under conditions where the disparity of the ribbon was ambiguous; with the KD, cylinder depth capture was obtained under conditions where the motion flow of the cylinder was in a direction parallel to that of the ribbon. These results demonstrate yet another similarity between KD and stereopsis. PMID- 10820603 TI - Porterfield and Wells on the motions of our eyes. AB - William Porterfield (ca 1969-1771) and William Charles Wells (1757-1817) conducted experimental investigations on eye movements related to accommodation, binocular vision, and vertigo. Porterfield gave a correct interpretation of Scheiner's experiment and invented an optometer to measure the near and far points of distinct vision. He also demonstrated the involvement of the crystalline lens in accommodation by examining vision in an aphakic person. Wells devised an alternative means of measuring the limits of vision and noted his own deterioration of sight with age; he studied the effects of belladonna on pupil size and accommodation. Their analyses of binocular visual direction contrasted Porterfield's view that perceived location was innately determined with Well's argument that visual direction was innate whereas visual distance was learned. Both Porterfield and Wells investigated the involvement of eye movements in binocular vision and in postrotary visual motion. Porterfield maintained that the eyes did not move following body rotation, whereas Wells, using an afterimage as stabilised retinal image, described the characteristics of postrotary nystagmus and their dependence on head orientation. Despite the neglect of Well's work, he should be considered as laying the foundations for the study of vestibular-visual interaction, even though the function of the vestibular system was not known at that time. PMID- 10820604 TI - Distractor ratio influences patterns of eye movements during visual search. AB - We examined the flexibility of guidance in a conjunctive search task by manipulating the ratios between different types of distractors. Participants were asked to decide whether a target was present or absent among distractors sharing either colour or shape. Results indicated a strong effect of distractor ratio on search performance. Shorter latency to move, faster manual response, and fewer fixations per trial were observed at extreme distractor ratios. The distribution of saccadic endpoints also varied flexibly as a function of distractor ratio. When there were very few same-colour distractors, the saccadic selectivity was biased towards the colour dimension. In contrast, when most of the distractors shared colour with the target, the saccadic selectivity was biased towards the shape dimension. Results are discussed within the framework of the guided search model. PMID- 10820605 TI - Perception of head orientation. AB - There are two visual components to gaze: head orientation and orientation of the eyes relative to the head. This study explores the accuracy with which subjects can discriminate head orientation when the eyes are centered in the head. Discrimination thresholds averaged 1.9 degrees of head rotation for base head orientations of 0 degree and 15 degrees, but discrimination was markedly poorer around a 30 degrees head orientation. Results were independent of spatial frequency and size over a 4-fold range. Neither negative contrast nor head inversion affected discrimination. Experiments dissociating the internal features from head outline revealed the presence of two main cues to discrimination: deviation of the head profile from bilateral symmetry, and deviation of nose orientation from vertical. Simulations show that model V4 units revealed in previous experiments with Glass patterns can extract the relevant head orientation information. The data are consistent with neurological data indicating a selective loss of face recognition in prosopagnosia with spared gaze discrimination. PMID- 10820606 TI - Perceptual learning in object recognition: object specificity and size invariance. AB - A series of four experiments measured the transfer of perceptual learning in object recognition. Subjects viewed backward-masked, gray-scale images of common objects and practiced an object naming task for multiple days. In Experiment 1, recognition thresholds decreased on average by over 20% over 5 days of training but increased reliably following the transfer to a new set of objects. This suggests that the learning was specific to the practiced objects. Experiment 2 ruled out familiarity with strategies specific to the experimental context, such as stimulus discrimination, as the source of the improvement. Experiments 3 and 4 found that learning transferred across changes in image size. Learning could not be accounted for solely by an improvement in general perceptual abilities, nor by learning of the specific experimental context. Our results indicate that a large amount of learning took place in object-specific mechanisms that are insensitive to image size. PMID- 10820607 TI - Every discrete, finite image is uniquely determined by its dipole histogram. AB - A finite image I is a function assigning colors to a finite, rectangular array of discrete pixels. Thus, the information directly encoded by I is purely locational. Such locational information is of little visual use in itself: perception of visual structure requires extraction of relational image information. A very elementary form of relational information about I is provided by its dipole histogram DI. A dipole is a triple, ((dx, dy), alpha, beta), with dx and dy horizontal and vertical, integer-valued displacements, and alpha and beta colors. For any such dipole, DI((dx, dy), alpha, beta) gives the number of pixel pairs ((x1, y1), (x2, y2)) of I such that I[x1, y1] = alpha, I[x2, y2] = beta, and, (x2, y2) - (x1, y1) = (dx, dy). Note that DI explicitly encodes no locational information. Although DI is uniquely determined by (and easily constructed from) I, it is not obvious that I is uniquely determined by DI. Here we prove that any finite image I is uniquely determined by its dipole histogram, DI. Two proofs are given; both are constructive, i.e. provide algorithms for reconstructing I from DI. In addition, a proof is given that any finite, two dimensional image I can be constructed using only the shorter dipoles of I: those dipoles ((dx, dy), alpha, beta) that have magnitude of dx < or = ceil((# columns in I)/2) and magnitude of dy < or = ceil((# rows in I)/2), where ceil(x) denotes the greatest integer < or = x. PMID- 10820608 TI - Global factors that determine the maximum disparity for seeing cyclopean surface shape. AB - A disparity gradient limit explains why the maximum amplitude of sinusoidal disparity gratings increases with decreasing disparity spatial frequency. It also explains why the largest disparity for binocular fusion (diplopia threshold) varies directly with stimulus element separation. Does a disparity gradient limit also apply to the detection of cyclopean shape? A previous study addressed this question and concluded that it does not. We examined this question by measuring the largest disparity amplitude (dmax) at which observers could judge the shape of cyclopean disparity gratings. We used trapezoidal, triangular, sinusoidal, and square wave gratings in order to dissociate the effects of disparity gradient and disparity spatial frequency. Gabor micropatterns were used to minimize potential scale-dependent interactions with luminance processing. Our results support a disparity gradient limit for cyclopean shape perception, with additional factors being involved at high disparity spatial frequencies. Combining the gradient limit hypothesis with lowpass disparity filtering describes the pattern of dmax for both smooth and discontinuous surface shapes. PMID- 10820609 TI - Summation between nearby motion signals and facilitative/inhibitory interactions between distant motion signals. AB - To explain the finding that motion assimilation was dominant between nearby motion signals while motion contrast between distant ones, a center-surround antagonistic mechanism was proposed [Nawrot & Sekuler (1990). Vision Research, 30, 1439-1451]. However, motion assimilation occurred not only between nearby signals but also between distant ones, suggesting the existence of a center surround non-antagonistic mechanism [Ido. Ohtani & Ejima (1997). Vision Research, 37, 1565-1574]. The present study was designed to provide direct evidence for the non-antagonistic mechanism, and to examine further the motion interactions which operate in different spatial scales. The nature of motion interaction between the test and the inducer was examined by varying the size, the number of frames, the frame duration and the inter-frame displacement of random-dot kinematograms. The results were consistent with the notion that there are three types of interactions in human motion processing; one is a summation process effective within nearby regions, and the other two are facilitative and inhibitory induction processes operating over larger spatial scales. Analysis of the results in terms of the Fourier components suggests that the facilitative and the inhibitory induction processes may be sensitive, respectively to the lower and the higher temporal frequency components of the stimulus. PMID- 10820610 TI - Linear combinations of signals from two lines of the same or different orientations. AB - Whereas the influence on the elevation of visually perceived eye level (VPEL) by two bilaterally symmetric, long (64 degrees-long), pitched-from-vertical lines in total darkness is only a little more than the average of the VPELs of the two lines measured separately [Matin & Li (1999). Vision Research, 39, 307-329], in the present experiments with 49 2-line combinations of seven orientations (-30 degrees to +30 degrees pitch), the VPEL for two short (12 degrees-long) lines equals the additive sum of the separate influences of the two lines. With one line at a fixed orientation, the slope of the VPEL-versus-pitch function with the second line variable equals the slope of the function when viewing one line alone, but is shifted from the 1-line-alone function by the magnitude of the VPEL of the fixed line. Both the near-averaging and the additivity are summarized by V(theta l, theta r) = k1 + k2 [V(theta l) + V(theta r)], where V(theta l) and V(theta r) are the 1-line VPELs for the pitches of the left and right lines, and V(theta l, theta r) is the 2-line VPEL; the slope constant k2 equals 0.5 for averaging, and 1.00 for simple additivity of the separate visual influences. Measured values are k2 = 0.99 and k2 = 0.61 for short and long lines, respectively. The shift of slope constant is determined by line length and not orientation: parallel and nonparallel lines follow the same rules of combination for short lines as they do for long lines. As for long lines, the short-line results are clear in showing that the visual influence on VPEL is controlled by an opponent-process mechanism. Although 'saturation-near-an-asymptote' along with opponency are required components of the interpretation for the basis of the combination of lines of different orientations and different lengths, they are not by themselves sufficient: All results conform to a neurophysiologically-based model [Matin and Li (1997b). Society for Neuroscience, 23, 175; Matin & Li, under review] that parallel processes feedforward signals from orientation-selective neural units in V1; the model accounts for the shift from additivity to near averaging with increase in line length as a consequence of the increased contribution of shunting. PMID- 10820611 TI - Prism induced accommodation in infants 3 to 6 months of age. AB - Convergence-accommodation, one of several cross-linkages in the oculomotor system is manifested by opening the accommodative feedback loop and increasing the vergence input. We elicited this response in human infants aged 3-6 months by placing a 15 delta prism (base-out) before one eye while they viewed a diffuse patch of light. Accommodation was measured and ocular alignment was confirmed with a video photorefractor. The convergence-accommodation response is therefore present during a time when blur driven accommodation and disparity vergence are maturing. The gain of convergence-accommodation (expressed as the stimulus CA/C ratio) appeared to be greater for infants than adults. PMID- 10820613 TI - Perceived heading during simulated torsional eye movements. AB - Observer translation through the environment can be accompanied by rotation of the eye about any axis. For rotation about the vertical axis (horizontal rotation) during translation in the horizontal plane, it is known that the absence of depth in the scene and an extra retinal signal leads to a systematic error in the observer's perceived direction of heading. This heading error is related in magnitude and direction to the shift of the centre of retinal flow (CF) that occurs because of the rotation. Rotation about any axis that deviates from the heading direction results in a CF shift. So far, however, the effect of rotation about the line of sight (torsion) on perceived heading has not been investigated. We simulated observer translation towards a wall or cloud, while simultaneously simulating eye rotation about the vertical axis, the torsional axis or combinations thereof. We find only small systematic effects of torsion on the set of 2D perceived headings, regardless of the simulated horizontal rotation. In proportion to the CF shift, the systematic errors are significantly smaller for pure torsion than for pure horizontal rotation. In contrast to errors caused by horizontal rotation, the torsional errors are hardly reduced by addition of depth to the scene. We suggest the difference in behaviour reflects the difference in symmetry of the field of view relative to the axis of rotation: the higher symmetry in the case of torsion may allow for a more accurate estimation of the rotational flow. Moreover, we report a new phenomenon. Simulated horizontal rotation during simulated wall approach increases the heading-dependency of errors, causing a larger compression of perceived heading in the horizontal direction than in the vertical direction. PMID- 10820612 TI - Visual and proprioceptive shifts in perceived egocentric direction induced by eye position. AB - The relation of three main effects of eye-position on perceived direction was investigated using a method of hand pointing in the horizontal plane: (1) Retinal eccentricity is overestimated with respect to the fovea by a constant factor of 2.6 degrees; (2) an extraretinal signal induces a shift in perceived visual direction (slope 0.12) that is opposite to the direction of eccentric gaze; and (3) the perceived position of the median plane of the head shifts toward the direction of eccentric eye-position (slope 0.23) while perceived trunk position remains unchanged. PMID- 10820614 TI - Spontaneous fluctuations in pupil size are not triggered by lens accommodation. AB - Fluctuations in pupil size and lens accommodation are measured concurrently under open loop conditions, constant illumination and far fixation. In 12/17 trials no correlation was measured between the fluctuations in pupil size and lens accommodation. For the remaining 5/17 trials no lag was observed between the changes in pupil size and lens accommodation indicating that this correlation does not arise as a consequence of a near response. These observations suggest that under conditions of constant illumination and far fixation, the supranuclear centers controlling the near response are not active. PMID- 10820615 TI - Color-specific depth mechanisms revealed by a color-contingent depth aftereffect. AB - Models of stereoscopic depth perception for both natural and random-dot images have focused mainly on the matching of achromatic features of binocular images. Recently, a growing body of research has investigated whether chromatic features can also contribute to the construction of stereoscopic depth. Here we present experiments yielding color-contingent depth aftereffects comparable in magnitude to those measured after adaptation to achromatic stimuli as evidence of neural mechanisms tuned to both color and depth. Furthermore, we report that the locus of the combined processing of color and depth is likely to lie beyond the site of binocular matching. PMID- 10820616 TI - The foveal 'crowding' effect: physics or physiology? AB - It has been known for some time that both foveal and peripheral visual acuity is higher for single letters than for letters in a row. Early work showed that this was due to the destructive interaction of adjacent contours (termed 'crowding' or contour interaction). It has been assumed to have a neural basis and a number of competing explanations have been advanced which implicate either high-level or low-level stages of visual processing. Our results suggest a much simpler explanation, one primarily determined by the physics of the stimulus rather than the physiology of the visual system. We show that, under conditions of contour interaction or 'crowding', the most relevant physical spatial frequency band of the letter is displaced to higher spatial frequencies and that foveal vision tracks this change in spatial scale. PMID- 10820617 TI - Form-deprivation myopia in monkeys is a graded phenomenon. AB - To shed light on the potential role of the phenomenon of form-deprivation myopia in normal refractive development, we investigated the degree of image degradation required to produce axial myopia in rhesus monkeys. Starting at about 3 weeks of age, diffuser spectacle lenses were employed to degrade the retinal image in one eye of 13 infant monkeys. The diffusers were worn continuously for periods ranging between 11 and 19 weeks. The effects of three different strengths of optical diffusers, which produced reductions in image contrast that ranged from about 0.5 to nearly 3 log units, were assessed by retinoscopy and A-scan ultrasonography. Control data were obtained from ten normal infants and three infants reared with clear, zero-powered lenses over both eyes. Eleven of the 13 treated infants developed form-deprivation myopia. Qualitatively similar results were obtained for the three diffuser groups, however, the degree of axial myopia varied directly with the degree of image degradation. Thus, form-deprivation myopia in monkeys is a graded phenomenon and can be triggered by a modest degree of chronic image degradation. PMID- 10820618 TI - Amodal completion and localization. AB - In a three-line vernier acuity task, a central line was localized between two aligned line segments. Accuracy did not improve when the segments formed a unified amodally completed percept. In a two-line vernier acuity task, a line was localized below a target line placed next to a physically segmented flank stimulus. Shifts in mean points of subjective equality suggested that amodally completed segments influenced localization. Previous conflicting findings are explained by a representation that is available early to influence perceived alignment. However, position tuning is poor. This representation may be realized physiologically by interpolation responses between amodally completed segments. PMID- 10820619 TI - Visual system's adjustments to illuminant changes: heuristic-based model revisited. AB - This study evaluates the effect of illuminant changes along the two post receptoral mechanisms: red-green (L - 2M) and yellow-blue (L + M - S). By means of a CRT colour monitor, Mondrian-type scenes were simulated and a series of asymmetric colour matches were made with five test illuminants. The standard objects comprising the scenes were simulations of surfaces under equal-energy illuminant and were selected according to lines of equal excitation of the red green and the yellow-blue mechanisms. Results show that observers' matches are well predicted by assuming affine transformations between test and standard illuminant conditions. The best linear fits derived from the data corroborates the previous heuristic-based algorithms [Zaidi Q. (1998) Journal of the Optical Society of America A. 7. 1767-1776] although some discrepancies were found. Results along red-green mechanism confirm that the significant effect of the illuminant is an additive change along this axis, while data for yellow-blue mechanism suggest that illuminant induces not only multiplicative changes along this axis but additive too. In addition, we found that memory factors involved in the experiment could influence the observers' matches and would be taken into account as responsible of the differences found between the yellow-blue and the red-green systems. PMID- 10820620 TI - A size illusion of the letter 'P'. AB - The perceived size of 'P' and 'p' is influenced by their vertical position relative to nearby letters. In the experiments reported here, we show that uppercase 'P' appears smallest when it is displaced downward toward the letter's lowercase position, whereas lowercase 'p' appears largest when it is near the uppercase position. Our results show that this P-illusion not only occurs in the presence of nearby letters, but is also found when 'P' is displaced relative to a nearby horizontal line. This type of size illusion seems to clearly occur only with 'P' and 'p', and we suggest that it is a result of learning to read and recognize letters in the English alphabet. PMID- 10820621 TI - The discrimination of abrupt changes in speed and direction of visual motion. AB - A random dot pattern that moved within an invisible aperture was used to present two motions contiguously in time. The motions differed slightly either in speed (Experiments 1 and 3) or in direction (Experiments 2 and 4) and the subject had to discriminate the sign of the change (e.g. increment or decrement). The same discrimination task was performed when the two motions were temporally separated by 1 s. In Experiments 1 and 2 discrimination thresholds were measured with motion durations of 0.125, 0.25, 0.5 and 1.0 s and mean speeds of 2, 4, 8, and 16 degrees/s. In Experiments 3 and 4 thresholds were measured with aperture widths of 5 and 20 cm. The discrimination of contiguous motions progressively deteriorated with decreasing duration and mean speed of motion. For the lowest value of duration the Weber fraction for contiguous speeds was more than three times as the Weber fractions for separate speeds. For the same low value of duration the thresholds for discrimination of direction of contiguous motions were only about 50% higher than the thresholds for separate motions. The Weber fraction for contiguous speeds was ca. three times higher with the smaller aperture than with the larger one, provided the ratio 'aperture width mean speed' (i.e. the lifetime of the moving dots) was less than 0.3 s. Aperture width did not affect the discrimination of direction of contiguous motions. The discrimination of contiguous motions is discussed together with the known data for detection of changes in speed and direction. It is suggested that both, detection of changes in speed and discrimination of the sign of speed changes, may be performed by a common visual mechanism. PMID- 10820622 TI - Spatial frequency tuned covariance channels for red-green and luminance-modulated gratings: psychophysical data from human adults. AB - Both chromatic and luminance-modulated stimuli are served by multiple spatial frequency-tuned channels. This experiment investigated the independence versus interdependence of spatial frequency channels that serve the detection of red green chromatic versus yellow-black luminance-modulated stimuli at low spatial frequencies. Contrast thresholds for both chromatic and luminance-modulated gratings were measured within 12 individual subjects using a repeated-measures design. Spatial frequencies ranged from 0.27 to 2.16 c/deg. A covariance structure analysis of individual differences was applied to the data. We computed statistical sources of individual variability, used them to define covariance channels, and determined the number and frequency tuning of these channels. For luminance-modulated gratings, two covariance channels were found, including one above and one below 1 c/deg [cf. Peterzell, & Teller (1996). Individual differences in contrast sensitivity functions: the coarsest spatial pattern analyzer. Vision Research, 36, 3077-3085]. For chromatic gratings, correlations between thresholds for most spatial frequencies were uniformly high, yielding a single covariance channel covering all but the highest spatial frequency tested. A combined analysis of both data sets recovered the same three covariance channels, and showed that detection thresholds for low-frequency red-green chromatic and luminance-modulated stimuli are served by separate, statistically independent processes. PMID- 10820623 TI - Spatial frequency tuned covariance channels for red-green and luminance-modulated gratings: psychophysical data from human infants. AB - This study concerns the spatial-frequency-tuned channels underlying infants' contrast sensitivity functions (CSFs) for red-green chromatic stimuli, and their relationship to the channels underlying infants' CSFs for luminance-modulated stimuli. Behavioral (forced-choice preferential-looking) techniques and stationary stimuli were used. In experiment 1. contrast thresholds were measured in 4- and 6-month-olds, using isoluminant red-green gratings with spatial frequencies ranging from 0.27 to 1.53 c deg. In experiment 2. contrast thresholds were measured in 4-month-olds. using both red-green and luminance-modulated gratings in the same low spatial frequency range. Covariance analyses of individual differences were performed. Experiment 1 revealed one dominant covariance channel for the detection of red-green gratings, with a second channel contributing to detection of the highest spatial frequencies used. Experiment 2 revealed two to three channels serving color and luminance: but surprisingly these channels were not statistically separable for luminance versus chromatic stimuli. Thus, covariance channels for color and luminance that are independent for adults [Peterzell & Teller (2000). Spatial frequency tuned covariance channels for red-green and luminance-modulated gratings: psychophysical data from human adults. Vision Research, 40, 417-430] are apparently interdependent in infants. These data suggest that for infants, detection thresholds for chromatic and luminance-modulated stimuli may be limited by common mechanisms. PMID- 10820624 TI - The development of symmetrical OKN in infants: quantification based on OKN acuity for nasalward versus temporalward motion. AB - We quantified OKN asymmetry in 140 normal infants, 3-24 months old, by varying spatial frequency to determine OKN acuity for temporal-to-nasal (T-N) versus nasal-to-temporal (N-T) motion. At all ages, OKN acuity was asymmetrical (better for T-N than for N-T motion) but the size of the asymmetry decreased from 3.2 to 0.7 octaves between 3-24 months, primarily because of improvements in OKN acuity for N-T motion. The results suggest that immaturities in the cortical pathways involved in OKN persist until at least 2 years of age. PMID- 10820625 TI - Psychophysical measurement of temporal modulation sensitivity in the tree shrew (Tupaia belangeri). AB - Temporal modulation sensitivity functions (MSFs) were measured behaviorally in three adult tree shrews (Tupaia belangeri). Shrews were trained to detect temporal sinusoidally-modulated full-field luminance variations in one of three stimuli, the two alternatives being static stimuli of equal size and time averaged luminance (34 cd/m2). Modulation depth was varied trial-by-trial using a modified staircase technique under ambient illumination of 16 lux. Threshold modulation depths were determined for five temporal frequencies ranging from 3.7 to 47 Hz. Results revealed temporal MSFs that peaked at 15 Hz with a low frequency roll-off and an extrapolated high-frequency cut-off beyond 50 Hz. These findings confirm the comparatively good temporal vision of Tupaia predicted by behavioral observations. PMID- 10820626 TI - Solutions at your fingertips. PMID- 10820627 TI - LASIK surgery. AB - The laser in situ keratomileusis (i.e., LASIK) procedure reshapes the surface of the cornea to focus visual images directly onto the retina, thereby improving visual acuity. In 1998, approximately 420,000 LASIK procedures were performed in the United States, and approximately two million people throughout the world already are benefiting from the LASIK vision correction procedure. PMID- 10820628 TI - Boyer's model of scholarly nursing applied to professional development. AB - Scholarship in nursing often is difficult to define and identify, but it is an important element in nursing. Scholars are needed in every practice setting to question the status quo and pursue fresh approaches to issues. This article examines Boyer's revised model of scholarship, which uses paradigms of discovery, integration, application, and teaching. Knowledge dissemination is emphasized as an important component for sharing information with fellow nurses and the public. Boyer's model of nursing scholarship provides institutions and OR nurse managers with a scholarly method for evaluating employee efforts. Strategies offered include professional development plans and employee portfolios. These applications encourage scholarly activities. PMID- 10820629 TI - Perioperative care of children in a transcultural context. AB - As part of a large, comparative study of how children are cared for in developed and developing countries' hospitals, health care professionals and parents were asked questions relating to their beliefs about parental presence during anesthesia induction and in postanesthesia care units. Children were not questioned. The researcher compared parents' (n = 957) and staff members' (n = 780) responses between developed and developing countries. Results indicate that parents and staff members in developed and developing countries responded significantly differently (P < .000001) to whether they believe parents or relatives should stay with their children until anesthesia takes effect. More staff members in developing countries responded that parents should be present (P = .007). Cultural constructions (e.g., class) are believed to influence strongly how care is delivered in developing countries studied. Cultural constructions were not as important in the developed countries. PMID- 10820630 TI - Parental participation in pediatric surgical care. AB - Hospitalization and surgery are stressful experiences for children and their parents. In recent years, pediatric health care has shifted toward family centered care that is based on close and continuous involvement of the child's family members. To shape and improve how pediatric care is delivered, health care providers need to know what children and parents need, expect, and experience. Such knowledge would enable the development of appropriate, systematic, and effective routines to optimize care for all children. Parental participation is beneficial to children, parents, and health care facilities, but it is dependent on the existence of effective routines to facilitate adequate communication among all parties. PMID- 10820631 TI - Selecting the perioperative patient focused model. AB - The Project Team on a Perioperative Model was charged with finding or creating a theoretical model for perioperative nursing. Team members evaluated 15 existing theories and decided to use the conceptual model developed by AORN's Data Elements Coordinating Committee. The Perioperative patient Focused Model is logical, and the concepts and principles are supported in practice environments. PMID- 10820632 TI - Maximizing nurses' advocacy role to improve patient outcomes. AB - Perioperative nurses are eminently situated, academically prepared, and professionally qualified to positively affect patient outcomes. Increasing efficiency and justifying the need for an RN in the perioperative milieu revolve around nurses' role as patient advocates. Perioperative nurses can take many measures to maximize this role, including reducing surgical site infections by enabling the timely administration of antimicrobial prophylaxis and by ensuring that surgical staff members realize the documented risks of removing hair from the surgical site. PMID- 10820633 TI - Nurses' contributions to the US space program. AB - Nurses have been engaged in aspects of aviation and space endeavors from their beginnings. Although no nurse has been chosen as a potential astronaut, nurses have contributed to the space program in many ways. Nurses and other health care providers use technology adapted from the space program. The future of extended space flight remains unclear, although work continues on completing a space station for extended stays on the moon and trips to Mars. Nurses may be one of the health care team members on these excursions. PMID- 10820635 TI - Internet resources for researching eye surgical procedures. PMID- 10820634 TI - Ensuring proper delegation to unlicensed assistive personnel. PMID- 10820636 TI - Methods to conduct focus groups and the moderator's role. PMID- 10820637 TI - Virtual instrumentation tools for real-time performance indicators: using PIVIT for data-driven decisions. PMID- 10820638 TI - Establishing maintenance intervals based on measurement reliability of engineering endpoints. AB - Methods developed by the metrological community and principles used by the research community were integrated to provide a basis for a periodic maintenance interval analysis system. Engineering endpoints are used as measurement attributes on which to base two primary quality indicators: accuracy and reliability. Also key to establishing appropriate maintenance intervals is the ability to recognize two primary failure modes: random failure and time-related failure. The primary objective of the maintenance program is to avert predictable and preventable device failure, and understanding time-related failures enables service personnel to set intervals accordingly. PMID- 10820639 TI - Effects of a wireless local area network (LAN) system, a telemetry system, and electrosurgical devices on medical devices in a hospital environment. AB - Concerns have been raised about interference of wireless local area network (LAN) systems and telemetry systems with medical devices in hospitals. The authors have investigated the susceptibility of 65 electromedical devices to a wireless LAN system and a telemetry system in preselected areas of a hospital. Testing was based on the American National Standards Institute Standard C63.18. The wireless LAN system operated at 2.42 GHz with an output power of 100 mW. The telemetry system operated at 466 MHz with an output power of 4 mW. Of the 65 devices tested, only two hand-held Doppler units, a Mini Doppler Model D900 (Huntleigh Healthcare Ltd) and a Ultrasonic Doppler Model 811 (Parks Medical Electronics, Inc.), were affected by the LAN system. Placed within 10 cm of the LAN system in standby mode, both units emitted periodic high-pitched beating sounds, which could be misinterpreted as normal beating sounds from the patient. These changed to random static noise during data transmission by the LAN. Under normal conditions of use, a LAN system would never be placed this close to a medical device. The quality of data transmission from the LAN system changed from "good" to "acceptable" in the colonoscopy room. This deterioration in transmission quality could have been caused by the lead shielding in the room. Electrosurgical devices operating at 0.5 to 1 MHz did not affect the LAN system at distances up to 3 m. None of the devices was affected by the telemetry system. These findings suggest that wireless LAN systems and telemetry systems can be acceptable for use in hospitals. Nevertheless, other systems should be tested on potentially susceptible devices by the hospital before use. PMID- 10820640 TI - Aeration time following ethylene oxide sterilization for reusable rigid sterilization containers: concentration of gaseous ethylene oxide in containers. AB - Because ethylene oxide (EO) gas is toxic to humans, restrictions have been imposed on its use for sterilization, specifying allowable levels of residual EO remaining in sterilized apparatus and materials. However, the aeration time that optimizes the removal of the remaining EO when a rigid sterilizing container is used for a vessel had not been identified. Therefore, polyvinyl chloride, which easily adsorbs EO, was placed in rigid sterilizing containers, and aeration was carried out after 1, 8, 12, 17, and 24 hours. After standard EO sterilization, the EO concentrations remaining in the air in the rigid containers were measured. The results indicate that a period of 17 hours of aeration is appropriate when a rigid sterilizing container is used. PMID- 10820641 TI - Ag-AgCl electrode noise in high-resolution ECG measurements. AB - The authors measured the noise and impedance from face-to-face Ag-AgCl electrode pairs, as well as the noise from Ag-AgCl electrodes placed on the human body surface, in the frequency band from 0.5 Hz to 500 Hz, which corresponds to high resolution ECG measurements. Electrode noise and electrode impedance were measured simultaneously to compare electrode noise with the thermal noise from the real part of electrode impedance. The results show that electrode noise depends on electrode area, electrolytic gel, the patient, and the placement site. In the frequency band from 0.5 Hz to 500 Hz, root-mean-square electrode noise is typically less than 1 microV for electrodes placed face-to-face and ranges from 1 microV to 15 microV for electrodes on the body surface. The noise spectral density increases at low frequencies as 1/fa and it is always higher than the thermal noise from the real part of the electrode impedance. There is a high correlation between electrode dc offset voltage and electrode noise. Thus, offset voltage measurements allow identification of noise from low-noise electrodes. PMID- 10820642 TI - Focus on medical errors provides clinical engineering opportunities. AB - Based upon the IOM report and the reaction it has already generated, as well as any new requirements that could emerge, we can expect to see increased pressure on health care to learn from its mistakes over the next few years. Clinical engineering professionals, who bring a breadth of experience and understanding in engineering, management and clinical principles to the table, have the ability to be an essential part of the team working to make the recommendations in the IOM report a reality. It is up to each of us to prepare ourselves to assume a prominent role within our hospitals as they tackle the challenge of reducing medical errors and improving patient safety. PMID- 10820643 TI - Second sourcing. Interview by Edward J. Snyder. PMID- 10820644 TI - Routing fundamentals, Part One: Distance vector routing. PMID- 10820645 TI - Oral delivery of HIV-protease inhibitors. AB - Strategies for optimizing the oral delivery of HIV-protease inhibitors draw from drug discovery efforts in molecular design, drug development tools in dosage formulation, and dosage regimen considerations in clinical medicine. This review outlines the evolution of these strategies for drugs that have been approved for human use, drug candidates still in development, and molecules that are no longer in development but from which valuable delivery information was obtained. Molecular design for obtaining desirable pharmacokinetics following oral administration primarily involved maximizing aqueous solubility and minimizing first-pass metabolism. Optimization of molecular design for oral drug delivery purposes is tempered by additional considerations for drug potency, toxicity, potential for interactions, and development of viral resistance. Strategies for improving oral bioavailability through dosage formulation use information from the effects of coadministered meals on drug plasma levels. Patient adherence to dosage regimens remains a major issue in assuring effective oral drug treatment and in preventing the development of resistance. Progress has been made in clinical studies where improved oral bioavailability and reductions in drug plasma level variability have been achieved with appropriate dosage regimen adjustment. PMID- 10820646 TI - Poly(ethylene glycol) conjugated drugs and prodrugs: a comprehensive review. AB - Low molecular weight Poly(ethylene glycol) (PEG) (< 20,000)-drug conjugates, prepared over a 20-year period, have been scrutinized and their properties and efficacy reviewed. No commercial products have thus far been reported for these types of compounds. However, during the past 5 years a renaissance in the field of PEG-(anticancer) drug conjugates has taken place, initiated by the use of higher molecular weight PEGs (> 20,000), especially 40,000, which is estimated to have a plasma circulating half-life of approximately 8-9 h. This recent resuscitation of small organic molecule delivery by high molecular weight PEG conjugates was founded on meaningful in vivo testing using established tumor models and has led to a clinical candidate. Recent applications of high molecular weight PEG prodrug strategies to amino-containing drugs are also detailed. PMID- 10820647 TI - Recent developments and emerging therapies for type 2 diabetes mellitus. AB - Most patients with type 2 (non-insulin-dependent) diabetes mellitus require pharmacotherapy, initially as monotherapy and subsequently in combination, as adjuncts to diet and exercise. Exogenous insulin is ultimately required in a substantial proportion, reflecting the progressive natural history of the disease. Sulphonylureas and biguanides have been employed for over 4 decades as oral antidiabetic agents, but they have a limited capacity to provide long term glycaemic control and can cause serious adverse effects. Thus, more efficacious and tolerable antidiabetic agents are required. Recent years have witnessed the introduction of agents with novel modes of action, that is, the alpha-glucosidase inhibitors acarbose and miglitol (which reduce postprandial hyperglycaemia) and the first of the thiazolidinedione insulinsensitising drugs--troglitazone and rosiglitazone. Although the former has been withdrawn in some countries due to adverse effects, another 'glitazone' pioglitazone is expected to be approved in the near future. Other recently introduced drugs include glimepiride and the meglitinide insulin secretagogue, repaglinide. Attention is also focusing increasingly on combination therapy using insulin together with sulphonylureas, metformin or troglitazone. Rapid-acting insulin analogues are now being used as alternatives to conventional insulins; their role in the management of type 2 diabetes mellitus is presently uncertain but reports of a reduced frequency of hypoglycaemia are encouraging. The development of new drugs aims to counter the principal metabolic defects of the disorder, respectively, relative insulin deficiency and insulin resistance. Novel classes of rapid-acting secretagogues under evaluation include the morphilinoguanide BTS 67582 and the meglitinides mitiglinide (KAD 1229) and senaglinide (A-4166). Succinate ester derivatives represent a potential novel approach to improving beta-cell function through enhancement of insulin biosynthesis and secretion. Enhancement of nutrient induced insulin secretion is a mechanism with several putative targets within the beta-cell; potentiators of insulin secretion include glucagon-like peptide-1 and its analogues, phosphodiesterase inhibitors and the imidazoline derivative PMS 812 (S 21663). The amylin agonist pramlintide slows gastric emptying and suppression of glucagon secretion. Non-thiazolidinedione insulin-sensitising agents include the gamma-receptor agonist G 1262570X (GG 570) and D-chiro inositol. Insulin analogues with prolonged action and inhaled insulin preparations are also under investigation. Insulin-mimetic agents include organic vanadium compounds. Whether newer agents will offer clinically relevant efficacy and tolerability advantages over existing therapies remains to be determined. PMID- 10820648 TI - Agents in development for type 2 diabetes. PMID- 10820649 TI - BTS 67582. PMID- 10820650 TI - Insulin aspart. Asp-B28, NovoRapid. PMID- 10820651 TI - Insulin glargine. Glargine, HOE 71GT15, HOE 71GT80, HOE 901. PMID- 10820652 TI - Insulin inhalation (Aradigm Corporation). NN 1998. PMID- 10820653 TI - Insulin inhalation (Pfizer/inhale therapeutic systems). HMR 4006. PMID- 10820654 TI - Mitiglinide. KAD 1229. PMID- 10820655 TI - PMS 812. S 21663. PMID- 10820656 TI - Pramlintide. AC 137, ACO 137, Normylin, Symlin, Tripro-amylin. PMID- 10820657 TI - Senaglinide. A 4166, AY 4166, Fastic, SDZ DJN 608, Starlix, Starsis, YM 026, Nateglinide. PMID- 10820658 TI - Agents in development for dementia syndromes in Japan. Summary and table. PMID- 10820659 TI - Azetirelin. YM 14673. PMID- 10820660 TI - Fasoracetam. LAM 105, NS 105. PMID- 10820661 TI - Nefiracetam. DM 9384, DZL 221, Translon. PMID- 10820662 TI - Perospirone. SM 9018. PMID- 10820663 TI - Sufoxazine. Lucelan, Metatone, Teniloxazine, Y 8894. PMID- 10820664 TI - TJ 960. PMID- 10820665 TI - Host/environment medicine and social realities. PMID- 10820666 TI - Research funding article is discussed. PMID- 10820667 TI - More on family medicine research funding article. PMID- 10820668 TI - Steelworker furnaces and families: from history and fiction to medicine. PMID- 10820669 TI - Research training for residents. PMID- 10820670 TI - Managing the difficult learning situation. PMID- 10820671 TI - Reflections from a rotation in Cambridge, England. PMID- 10820672 TI - Accuracy of electrocardiogram reading by family practice residents. AB - OBJECTIVES: This study evaluated the electrocardiogram (EKG) reading skills of family practice residents. METHODS: A multicenter study was carried out to evaluate the accuracy of EKG reading in the family practice setting. Based on the frequency and potential for clinical significance, we chose 18 common findings on 10 EKGs for evaluation. The EKGs were then distributed to residents at six family practice residencies. Residents were given one point for the identification of each correct EKG finding and scored based on the number correct over a total of 18. RESULTS: Sixty-one residents (20 first year, 23 second year, and 18 third year) completed readings for 10 EKGs and were evaluated for their ability to identify 18 EKG findings. The median score out of 18 possible points for all first-, second-, and third-year residents was 12, 12, and 11.5, respectively. Twenty-one percent of residents did not correctly identify a tracing of an acute myocardial infarction. Data analysis showed no statistically significant difference among the three groups of residents. CONCLUSIONS: We evaluated the accuracy of EKG reading skills of family practice residents at each year of training. This study suggests that EKG reading skills do not improve during residency, and further study of curricular change to improve these skills should be considered. PMID- 10820673 TI - Rejecting family practice: why medical students switch to other specialties. AB - BACKGROUND AND OBJECTIVES: Medical schools have been encouraged to increase the number of primary care graduates. This study determined the proportion of medical students who change specialty preference during the clinical years and explored how ultimate choice is affected by perceptions of medical specialties acquired during this period. METHODS: A survey was mailed to 397 graduating medical students at the University of California, San Francisco (UCSF) after the National Resident Matching Program Match and before graduation in 1996, 1997, and 1998. RESULTS: The response rate was 81% (320/397). Of 41 respondents who reported that family practice had been their first specialty choice prior to beginning clinical rotations, only 15 (37%) eventually matched in family practice. Comparable numbers for internal medicine and pediatrics were 50% and 69%. Students rejecting family practice were more likely than their colleagues rejecting other specialties to cite insufficient prestige, low intellectual content, and concern about mastering too broad a content area as reasons. CONCLUSIONS: At UCSF, family practice retains fewer interested students than other primary care specialties. To reverse this trend, schools such as UCSF need to raise the prestige of family practice and counter concerns about its intellectual content being impossible to master. PMID- 10820674 TI - Responding to patients' emotions: important for standardized patient satisfaction. AB - BACKGROUND AND OBJECTIVES: Several experts have emphasized the need to respond to a patient's emotions as an essential component of effective medical interviewing. This study examined the relationship of faculty observers' scores of students' performance in standardized patient (SP) interviewing stations in a family medicine clerkship objective structured clinical examination (OSCE) with SP satisfaction measures. METHODS: The faculty observers scored students in the following performance domains: 1) interviewing skills, 2) negotiating the diagnosis or plan, 3) gathering case-specific content information, 4) responding to the patient's emotions, and 5) student's overall performance. Pearson Product Moment correlations were calculated for each of these domains and the Standardized Patient Satisfaction Questionnaire (SPSQ) scores. RESULTS: There were moderate correlations between the SPSQ score and the overall OSCE score (.45) and the response to patient's emotions skill score (.36). The faculty observer's response to patient's emotion score correlated highly with the student's overall OSCE score (.75). CONCLUSIONS: A student's ability to respond to the patient's emotions appears to be an important skill for successful medical interviewing from both the faculty observer's perspective and the SP's perspective. Results also demonstrate that the SP's perspective is similar, but not identical, to the faculty observer's perspective, suggesting that SPs have an important evaluative role in student assessment. PMID- 10820675 TI - US medical schools and the rural family physician gender gap. AB - BACKGROUND: Women comprise increasing proportions of medical school graduates. They tend to choose primary care but are less likely than men to choose rural practice. METHODS: This study used American Medical Association masterfile data on 1988-1996 medical school graduates to identify the US medical schools most successful at producing rural family physicians and general practitioners of both genders. RESULTS: The number of listed rural female family physician or general practitioner graduates among schools ranged from 0-27 (0% to 4.4% of each school's 1988-1996 graduates). There were approximately twice as many male as female rural family physicians and general practitioners. Publicly funded schools produced more rural female family physicians and general practitioners than their privately funded counterparts. CONCLUSIONS: Our findings suggest that a few schools, most of them public, may serve as models for schools that aim to train women who later enter rural practice. PMID- 10820676 TI - Improving pneumococcal vaccination rates: a three-step approach. AB - BACKGROUND: Streptococcus pneumoniae is the cause of significant preventable morbidity and mortality each year, especially among the elderly. This study evaluated the effectiveness of a three-step strategy to improve pneumococcal vaccination rates in patients ages 65 and older. METHODS: We obtained a baseline vaccination rate by chart review of a randomized list of patients ages 65 and older. Interventions that included physician education, a system of office prompts, and patient education were instituted for 1 year. We performed a second chart review to determine the success of the intervention. RESULTS: Vaccination rates increased significantly from 56.7% to 75.8%. No significant difference in initial vaccination rates between men and women was found. Although rates improved for both men and women, only the increase in vaccination for men was significant. CONCLUSIONS: This study shows that an easy and inexpensive intervention can substantially impact pneumococcal vaccination rates. Specifically, a three-step approach that combines provider education, office prompts, and patient education has proven effective. PMID- 10820677 TI - Israel under threat of biological warfare--the reactions of our patients during the 1998 Persian Gulf crisis. AB - BACKGROUND AND OBJECTIVES: During February 1998, there was heightened tension in the Persian Gulf area. Iraq's ruler, Saddam Hussein, threatened Israeli citizens with the use of biological weapons. This study observed the use of health care services by patients visiting our clinics during this period. METHODS: During the period of February 17, 1998, through March 1, 1998, 12 family physicians classified all clinic visits as "related" or "unrelated" to the crisis. The participating clinics were situated in (A) high-risk and (B) low-risk areas, according to their location as related to missile hits during the 1991 Gulf War. RESULTS: A total of 1,841 visits were recorded during the crisis period (February 17-23). There were 934 visits in Area A and 907 in Area B. Overall, 194 visits related to the crisis were recorded, of which 155 were in Area A. The rate of crisis-related visits was higher in Area A (16.6% versus 3.4%). There were more visits for authorizations for special masks (8.2% versus .8%), requests for information (3.9% versus 1.3%), and tranquilizers (2.1% versus .5%). In the period subsequent to the agreement reached on February 24, crisis-related visits declined sharply in both areas. CONCLUSIONS: During the crisis, there was an increase in visits related to the threat of biological warfare. The phenomenon was more pronounced in areas that had sustained missile hits during the 1991 Gulf War. PMID- 10820678 TI - Family practice residency training: can we make it better? PMID- 10820679 TI - Creative potential, creative achievement, and personal growth. AB - We tested the idea that young people who have creative potential (are complex and unconventional) increase in intrapsychic awareness as they mature but often have difficulty with psychosocial growth, especially the development of a cohesive identity; and that for some women commitment to creative work solves the problem of psychosocial integration and leads to creative achievement. In a longitudinal sample of 109 women, these ideas were supported: Creative potential and creative achievement were both associated with intrapsychic growth but only creative achievement was associated with psychosocial growth. Regression analyses showed that the development of a cohesive identity from early to middle adulthood added to and interacted with creative potential in the prediction of creative achievement. PMID- 10820680 TI - A behavioral genetic analysis of the relationship between the socialization scale and self-reported delinquency. AB - This investigation examined the genetic (A), and shared (C) and nonshared (E) environmental variance contributions to the relationship of self-reported delinquency (as measured by the "Delinquent Behavior Inventory" [DBI; Gibson, 1967]) to the Socialization (So) scale of the California Psychological Inventory using univariate and bivariate structural equation models. The scales were administered to 222 male (145 monozygotic; 77 dizygotic) and 159 female (107 monozygotic; 52 dizygotic) 16- to 18-year-old same-sex twin pairs. Principal components analysis with varimax rotation revealed three interpretable So factors representing family/home environment, self-concept, and behavioral control. Univariate modeling suggested sex differences in etiological influences associated with individual differences in most scales. The bivariate ACE model fit the data, suggesting that the covariance between the So scale and self reported delinquency owes in part to shared etiological factors. PMID- 10820681 TI - Personal goals in social roles: divergences and convergences across roles and levels of analysis. AB - Most contemporary personal goal research aggregates across goals, perhaps masking important differences between goals. We assessed this risk by examining both similarities and differences between the goals that participants pursued in five important social roles. Previous relevant findings (Cantor, Norem, Niedenthal, Langston, & Brower, 1987) and self-determination theory (Deci & Ryan, 1985) were used to predict between-role differences in goal appraisal dimensions. Although theoretically meaningful differences were found across child, employee, romantic, friendship, and student goals, and also across within- and between-subject levels of analysis, all goals were essentially the same in one important way: Making longitudinal progress in them predicted positive change in accompanying role circumstances and role-satisfaction (excepting friendship goals). This indicates that researchers do not necessarily lose information by aggregating, and affirms that goal-attainment is generally desirable. PMID- 10820682 TI - Comparing personality scales across time: an illustrative study of validity and consistency in life-span archival data. AB - The goals of this study were: (a) to examine whether personality scales, meaningful in contemporary terms, could be derived from archival data; and (b) to use these scales to aid our understanding of the relation of personality to mortality. NEO PI-R data and a battery of archival items, taken from Terman's Life Cycle Study, were collected on two new samples (sample 1 mean age = 11.9, n = 167; sample 2 mean age = 22.2, n = 203). Measurement invariance of the archival scales was assessed, and validity was examined using both rational analyses and associations with the Five Factor Model. It was demonstrated that interpretable scales can be derived from 50- to 70-year-old archival data. The archival adult personality data were then used to predict mortality. Conscientiousness remains the strongest personality predictor of longevity. Criteria for establishing the validity of archivally derived scales are suggested. PMID- 10820683 TI - Free-response self-discrepancies across, among, and within FFM personality dimensions. AB - An approach for personality-based self-discrepancy (PBSD) measurement is proposed, whereby self-beliefs and incongruities among their contents are assessed with respect to five-factor model (FFM) semantic dimensions. Selves Questionnaire attributes from 191 college students were coded against L. R. Goldberg's (1990) FFM factor analysis to construct personality scores for actual, ideal, and ought self domains, as well as several PBSD indices. Multivariate analyses were conducted to test self-discrepancy and personality-structure hypotheses, and to demonstrate this strategy's operational flexibility. Profile analyses indicated that empirical self-discrepancies depend upon whether and how personality structure is incorporated. Methodological alterations influenced self discrepancy findings negligibly. Initial evidence for PBSD construct validity and predictive specificity is presented, and the approach's implications, advantages, and extensions are discussed. PMID- 10820684 TI - Spirituality: description, measurement, and relation to the five factor model of personality. AB - The present article focused on the development and measurement of a factor model of the expressions of spirituality. Study 1 (N = 534) involved the use of factor analysis to examine the latent factor structure in a sample of 11 measures of spiritual constructs. Study 2 (N = 938) focused on the replication of Study 1 results and on the construction and initial validation of an instrument to operationalize the factor model of spirituality. Results indicate that at least 5 robust dimensions of spirituality underlie the spirituality test domain. These dimensions were labeled Cognitive Orientation Towards Spirituality (COS), Experiential/Phenomenological Dimension (EPD), Existential Well-Being (EW-B), Paranormal Beliefs (PAR), and Religiousness (REL). The measure developed, named the Expressions of Spirituality Inventory (ESI), takes the form of a 98-item instrument that generated scores demonstrating satisfactory reliability and adequate initial validity. Examination of the relation of spirituality to the Five Factor Model (FFM) as measured by the NEO Personality Inventory-Revised revealed that the dimensions of the FFM appear to differentially relate to the major elements of spirituality but are nevertheless conceptually unique, pointing to the possible existence of major aspects of personality not represented in the FFM. PMID- 10820685 TI - Hope and dysphoria: the moderating role of defense mechanisms. AB - It was hypothesized that the adaptive value of hope cognitions would be dependent upon the quality of an individual's defense style. Undergraduate students completed measures of hope, defense mechanisms, and dysphoria in two studies. As predicted, defense mechanisms significantly moderated the relation between hope and dysphoria. In addition, both hope and defense mechanisms predicted dysphoria as main effects. Individuals who had low hope and an immature defense style had particularly high levels of dysphoria. Low hope was not maladaptive for individuals with a mature defense style, suggesting that a subtype of low hope ("defensive hopelessness") may exist that is analogous to defensive pessimism. Individuals with high hope had low levels of dysphoria regardless of defense style. Overall, the present study suggests that an integration of psychodynamic and cognitive perspectives on hope may be productive. PMID- 10820686 TI - Fragile self-esteem in children and its associations with perceived patterns of parent-child communication. AB - We examined the extent to which 11- to 12-year-old children's (N = 174) self esteem (SE) stability and level related to their perceptions of various aspects of parent-child communication. Compared to children with stable SE, children with unstable SE reported that their fathers were more critical and psychologically controlling, and less likely to acknowledge their positive behaviors or to show their approval in value-affirming ways. Likewise, children with low SE reported that their fathers exhibited these qualities to a greater extent than did children with high SE. In addition, fathers of children with stable high SE were viewed as especially good at problem solving. Children's SE level related to perceptions of mothers' communication styles very similarly to how it did with fathers'; with respect to SE stability, however, relationships were generally less consistent and frequently absent. Discussion centered on the role of parent child communication in promoting unstable SE. PMID- 10820687 TI - Thinking ahead: complexity of expectations and the transition to parenthood. AB - This study examined the integrative complexity of thinking in individuals making the transition to parenthood, and the relationship between complexity and adjustment during this period. Sixty-nine couples were interviewed 3 months before their babies were born, and 6 months after the birth. The prenatal interview focussed on individuals' expectations about what it would be like being a parent; the postnatal interview focussed on individuals' actual experiences as parents. In addition, participants completed measures of depression, self-esteem, and marital satisfaction after each interview, and a measure of stress after the 6-month postnatal interview. Both men and women demonstrated a significant increase in the complexity of their thinking from the prenatal to the postnatal interview, with women demonstrating higher levels of complexity at both times. In addition, women with more complex expectations demonstrated better adjustment after their babies were born than did women with simpler expectations; these results were not obtained for men. Results are discussed with regard to the way in which thinking about the self changes as one negotiates major life transitions, and the way in which complex thinking can help counter some of the stresses that individuals may experience at these times. PMID- 10820688 TI - The fruits of their labors: a longitudinal exploration of parent personality and adjustment in their adult children. AB - This longitudinal study of several aspects of parenting examines how children "turn out" as adults. Ratings of adjustment (educational and occupational attainment as well as social and emotional maturity) in young adult children (aged 25-37) were made on the basis of information from 64 mothers who were participants in a longitudinal study of women's lives. As hypothesized, mothers who were demanding yet responsive, sensitive, and psychologically mature at ages 27 and 43 had children with higher overall adult adjustment scores. Other factors correlated with adjustment included the mother's long-term commitment to being a wife and mother, and the cohesiveness of the home environment. In their 50s, characteristics of parents associated with adult child adjustment were different for men and women (competence in women and forcefulness and individuality in men). Divorce was not a negative factor, and mothers' paid work (after age 27) was positive at a trend level. PMID- 10820689 TI - Worries and values. AB - Relations of individuals' value priorities to their worries are investigated in seven samples from four cultural groups (N = 1,441). A social-cognitive analysis suggests that value priorities influence worries by increasing attention to and perception of threats to valued goals. On this basis, we generate hypotheses relating two types of worries, micro (about self and its extensions) and macro (about society and world), to 10 types of values. As predicted, giving priority to self-transcendence values (universalism and benevolence) is associated with low micro and high macro worry, whereas giving priority to self-enhancement values (power, hedonism, and--to a lesser degree--achievement) is associated with high micro and low macro worry. Meaningful associations are also found for other values. Values account for substantially more variance in macro than in micro worries. PMID- 10820690 TI - Self-pluralism: assessment and relations to adjustment, life changes, and age. AB - Individuals differ in the degree to which they see themselves as behaving and feeling similarly or differently in different situations and at different times. This dimension of the self-concept, which is conceived to extend from strong constancy of the self-concept at one extreme to marked variability at the opposite pole, is designated as self-pluralism. An inventory for the assessment of degree of self-pluralism--the Self-Pluralism Scale (SPS)--is presented, and the correlations of this scale with other relevant personality measures are reported. Several studies utilizing the SPS are presented. The results indicate that self-pluralism is negatively related to psychological adjustment, positively related to long-term real-life variability, and negatively related to age. PMID- 10820691 TI - Extrinsic value orientation and "the tragedy of the commons". AB - Two studies examined the effect of Extrinsic Value Orientation (Kasser & Ryan, 1993, 1996) upon harvesting strategies and personal profit within commons dilemmas, in which individual and group interests can be at odds. At an individual or within-group level of analysis, extrinsically oriented persons (who value money, fame, and popularity) harvested more than intrinsically oriented persons (who value self-acceptance, intimacy, and community). However, a counteracting group-level effect was found such that groups with a greater number of extrinsic members harvested less on average than did groups with more intrinsic members, because their commons did not last as long. As a result, even excessive harvesters within extrinsic groups did no better than did self restrained harvesters within intrinsic groups. Supplementary analyses indicate that extrinsic values are associated with acquisitiveness regarding resources, more so than apprehension regarding others' acquisitiveness. PMID- 10820692 TI - Public health and mental health. PMID- 10820693 TI - What every nurse needs to know about ... juvenile firesetters. A report of the Juvenile Firesetter Intervention Project. AB - With the number of juvenile firesetters growing each year and the amount of psychological and financial damage they create, it is essential that clinical professionals are aware of the characteristics surrounding juveniles who set fires. This study had two primary purposes: to educate nurses and mental health practitioners about the etiology and treatment of juvenile firesetters and to discuss a number of exploratory predictors, including age, gender, aggression, internalizing behaviors, level of deviancy, family dynamics, and sociability. Certain individual and environmental characteristics relate to varying levels of damage caused by the fire and the presence or absence of recidivistic behaviors. Firesetters are a group widely seen in the general population but not often identified or studied by clinical professionals. Therefore, this article has implications for the clinical practice of nurse practitioners and others who come into contact with this clinical population. PMID- 10820694 TI - Psychiatric patients' perceptions of waiting time in the psychiatric emergency service. AB - Recent customer service surveys have indicated that patients in the Psychiatric Emergency Service (PES) are increasingly dissatisfied with the "waiting" time connected with expanded services. This study examined the impact of six interventions that altered the environment of waiting patients and their resulting perceptions of time spent in PES. The interventions were communication (three styles), relaxing music, educational videos, and recreational activities. Each intervention was conducted for 7 days, followed by 7 days of no intervention. Patients completed a Time Assessment Tool that measured expectations of, perceptions of, and satisfaction with waiting time. These data were compared to actual time, as reflected on the log maintained in the PES. In general, environmental interventions reduced patients' perceived time, compared to no intervention. Significantly, interacting with patients with caring and concern consistently resulted in a reduced perception of waiting time, compared to the other interventions. PMID- 10820695 TI - Lunacy revisited. The influence of the moon on mental health and quality of life. AB - The idea that the stars and planets may influence human health and behavior can be traced to at least Roman times, and research suggests a high proportion of health professionals continue to hold this belief. Nevertheless, evidence for the supposed influence of the moon on human behavior has proved particularly elusive, and research has tended to suffer from weaknesses in methodology and data analysis. This article reports findings drawn from a re-analysis of data from a research study into the functioning of a sample of mentally ill people living in the community. The mental health and quality of life of a sample of 100 people were assessed on four occasions during a 30-month period. Data were aggregated to represent the span of one lunar month, with scores being allocated to the relevant week of the lunar cycle during which each assessment was made. Comparison of mean values across the weeks of the lunar cycle was preformed using the ANOVA, Results showed significant change at the time of the full moon only in subjects with a diagnosis of schizophrenia (n = 56), where deterioration was observed in three areas of psychopathology and one area of quality of life. Some implications for nursing practice are discussed, and it is suggested that future research into the possibility of a lunar effect on human life should focus on the direct measurement of functioning in people with schizophrenia. PMID- 10820697 TI - Dramatic rise in the number of preschoolers taking psychiatric drugs. PMID- 10820698 TI - Parents' choice: custody vs. treatment. PMID- 10820696 TI - Maximizing learning outcomes by videotaping nursing students' interactions with a standardized patient. AB - 1. Videotaping is a beneficial instructional methodology to help undergraduate students develop awareness of their strengths and opportunities for growth in therapeutic interactional skills. 2. Educational interventions with undergraduate students provide an opportunity to involve doctoral students in vital instruction, research, and knowledge dissemination through publication and presentation. 3. Allowing novice nurses to view their interactional skills with a simulated patient can reduce the anxiety they experience in first therapeutic encounters with assigned patients. 4. Continuous quality improvement in educational methodology is dependent on listening carefully to students who have invaluable feedback to offer and making appropriate correction as indicated. PMID- 10820699 TI - Differences in men's and women's brains responsible for differences in schizophrenia. PMID- 10820700 TI - [Impaired motility. Why strabologists are unwilling to move]. PMID- 10820701 TI - [Employing a medical assistant? Which?]. PMID- 10820702 TI - [Specular microscopy follow-up of endophthalmitis after cataract operation]. AB - INTRODUCTION: Since the first reports of specular microscopic photographs in vivo of the corneal endothelium, endophthalmitis is said to damage endothelial cells irreversibly. PATIENTS AND METHOD: We controlled 29 eyes 1 to 10 years after endophthalmitis following cataract surgery using specular microscopy. We tried to find out, if endophthalmitis leads to significant endothelial damage and we wanted to describe the in-vivo-cytological follow-up of the implanted lenses. RESULTS: Endothelial cell-density of the eyes with endophthalmitis (n = 29) was 2733/mm2 (+/- 680). In those eyes receiving cataract-surgery without endophthalmitis in the fellow eye (n = 14), the endothelial density was 2851/mm2 (+/- 360). If the fellow eye has had no cataract surgery (n = 13), cell density was 3110/mm2 (+/- 750). Hypopyon-iritis after cataract-surgery did not decrease endothelial cell count significantly more than cataract-surgery without hypopyon iritis (p > 0.05). At the last control at least one year after surgery, 36% of all IOLs were free of any cellular deposits. There was no case of a foreign-body reaction. CONCLUSION: In our patients, postoperative endophthalmitis has not led to significant endothelial damage. In none of our patients, endophthalmitis has led to chronic foreign-body reaction against the implant or to granulomatous uveitis. Specular microscopy of the corneal endothelium and of the implanted lens may help to differentiate in the case of postoperative inflammation between an infection and a foreign-body-reaction. In the case of a sterile foreign-body reaction, there are no inflammatory cells on the endothelium or in the anterior chamber. PMID- 10820703 TI - [25 years Cardona keratoprosthesis after severe chemical eye burns--long-term outcome of 4 eyes]. AB - BACKGROUND: A keratoprosthesis should be implanted only in such eyes, in which a risk-keratoplasty (HLA-typified) was unsuccessful. However, the keratoprosthesis is an ultima ratio procedure. PATIENTS AND METHODS: 25 years ago a modified keratoprosthesis according to Cardona was implanted in 2 patients in 4 eyes with very heavy alkali burns. Complications are described in a 25-years follow up in all of the 4 eyes. The loss of visual acuity was the first alarm sign for each complication. Reversible epicorneal complications are: epithelium over-running in front of the prosthesis, retraction of mucosa, lyophilized sclera including fistulation and dislocation or extrusion of the prosthesis. The opposite of the epithelium over-running was the development of a retro-prosthetic membrane as a sign for over-running inside. Severe (intraocular) complications were: macular oedema and choriodal detachment in bulbar hypotension, vitreal haemorrhage, retinal detachement and exacerbation of a secondary glaucoma. An endophthalmitis and later a phthisis bulbi were observed due to a permanent fistulation. RESULTS: Exactly after 25 years the visual acuity is in both patients: patient 1: 0.6 p and amaurosis; patient 2: 1.3 p and 0.8 p. The visual field is limited by the keratoprosthesis usually to 30 to 40 degrees, in our both eyes with secondary glaucoma the visual field was reduced to 10 and 25 degrees. CONCLUSIONS: Bad auspices bring many complications and many operations. The patients should be sent back to the surgeons very early. Besides, a psychological guidance is important for the patients. PMID- 10820704 TI - [Clinical anophthalmos. Cosmetic outcome after 2 years therapy with an orbital expander for stimulating orbital growth]. AB - BACKGROUND: Congenital anophthalmos is a rare condition in which intervention in an early age can stimulate orbital expansion. The therapeutic goal was to allow retention of a suitable prosthesis and to maximise facial symmetry. METHODS AND PATIENTS: We report on 3 anophthalmic newborn male patients, of which one presented with microphthalmos in the other eye as well as associated systemic disorders. Solid moulded shapes were adapted and manufactured out of methylmethacrylate to increase expansion of orbital soft tissue and bone. The moulding of the orbita was performed by silicone made of two components under general anaesthesia during the first two years of life. The following day the solid shapes were fitted in the outpatient department. They were increased in size when they started to rotate within the orbit or when the eyelids were relatively loose around the solid shape. The initial insertion of the orbital expander was performed in the first weeks after birth and was repeated every 3 to 6 weeks within the first year of life. The degree of orbital expansion was determined by measurements of the horizontal eyelid length, the volume of the solid shapes, by measuring the volume in cm3 of water displaced by the shape after submerging it in a cylinder of water and assessment of the cosmetic aspect. RESULTS: Within 8 months of therapy a horizontal eyelid lengthening to 10 mm and an increase of the volume of the orbital expander from 1.5 cm3 to 6 cm3 was achieved in patient 1. After 20 months of therapy patient 2 showed symmetry of the face and the horizontal eyelid length. In patient 3 therapy started 3 years late and after 2 years only a horizontal eyelid lengthening of 3 mm and a suboptimal symmetry of the face was achieved. CONCLUSION: Our own experience suggests that early prosthetic fittings are necessary for an ideal cosmetic outcome. Motivation and cooperation of the parents is an important factor to achieve optimal results. PMID- 10820705 TI - [Changes in the lens epithelium of diabetic and non-diabetic patients with various forms of opacities in senile cataract]. AB - BACKGROUND: The crucial role of the lens epithelium with respect to cataractogenesis has to be further evaluated. In this prospective clinical study, morphological characteristics of human lens epithelium in type-II diabetics and nondiabetics were examined and compared. MATERIALS AND METHODS: 30 (16 female, 14 male, mean age 73.0 years) of the overall 59 patients with age-related cataract suffering from type-II diabetes were assigned to group I and 29 nondiabetics (16 female, 13 male, mean age 68.2 years) to group II. Age, gender, type of cataract and the blood parameters fasting blood sugar and glycolized hemoglobin were further parameters. The different types of opacities (LOCS II) were summarized into 4 groups. After surgery the collected hematoxylin-eosin stained anterior central lens capsules with attached lens epithelial cells were analyzed by light microscope for the cell parameters cell density (morphometry), nucleus area (A0), nucleus volume (V), cell area (A) and nucleus-plasma-ratio. RESULTS: The mean cell density in type-II diabetics (group I) is 3691 +/- 346 cells/mm2 and in nondiabetics (group II) 4162 +/- 504 cells/mm2, respectively (p = 0.001). The total female mean cell density (4036 +/- 525 cells/mm2) was not significantly higher than the male (3788 +/- 412 cells/mm2). A decrease of the mean cell density could be attributed to age only in the nondiabetic group. With regard to the type of cataract the posterior subcapsular cataract shows the lowest mean cell density (3620 +/- 333 cells/mm2) and the nuclear cataract (4250 +/- 513 cells/mm2) the highest, respectively. The medium nucleus area and -volume and cell area are in the type-II diabetic group significantly larger than in nondiabetics. With regard to the type of cataract the posterior subcapsular opacity has the significantly largest values. The medium nucleus-plasma-ratio in type-II diabetics is lower than in nondiabetics and decreased with age. CONCLUSIONS: The significantly lower mean cell density in type-II diabetics compared with nondiabetic eyes and in posterior subcapsular cataracts in comparison with nuclear and cortical cataracts seem together with the other morphological cell characteristics to be due to the cataractogenic influence of diabetic metabolic disorder on the lens epithelium, especially in cases of posterior subcapsular opacity. These may be some clues for the primary cataractogenic importance of the lens epithelium in type-II diabetes. PMID- 10820706 TI - [Tubingen 1895-1899. Incidence, diagnosis and therapy of selected optic nerve, uveal and retinal diseases 100 years ago]. AB - BACKGROUND: The records of the University Eye Clinic are almost completely preserved at the historical archive of the University of Tubingen since 1865 and shell now be reduced. MATERIALS AND METHODS: 150 records from the historical archive of the University of Tubingen were selected. We were interested in those patients treated for the first time at the University Eye Clinic between 1895 and 1899. Only diseases of the optic nerve, the uveal tract and the retina were considered. The records were distributed into groups following historical classification and statistics of the same period. These statistics were also used to compare our selection with all patients treated. For information about frequency, diagnostics and therapy a questionnaire was developed. RESULTS: Glaucoma: The data of the subgroup "Glaucoma simplex" showed high correspondence with the historical data regarding the frequency (24.3 vs. 25.5%). The data of the subgroup "Glaucoma secundaria" differed (34.5 vs. 25.5%). Testing the visual acuity was the first step of examination here and in all groups (100%). Measurement of intraocular pressure with digital palpation followed (86.3%). First of all Eserin eye drops were applicated (76.5%). Iritis: The frequency of acute iritis was 16.8 vs. 25.0%, and of chronic iritis 59.6 vs. 52.2%. As typical symptoms ciliar injections, fixed anisocore pupil and exudation were often seen (80.0%). Chronic iritis showed as typical symptom posterior synechia (80.9%). Atropin eye drops were firstly applied in acute iritis (100%), chronical iritis was treated by iridectomy (57.1%). Myopia: In all subgroups our data differed from the historical data. Staphyloma posticum was the characteristic diagnosis (65.8%). Myopia was treated by decision of the lens. Retinal detachment: The frequency 37.2 vs. 28.0%. In nearly all cases retinal detachment was described after ophthalmoscopic examination (92.8%). The first therapeutical step was to keep the patients staying in bed (53.8%). CONCLUSIONS: As expected, diagnostic and therapeutical options were strongly limited in Tubingen 100 years ago. Nevertheless, therapeutic successes were achieved in certain cases. PMID- 10820707 TI - [Tilorone-induced functional changes in the rat retina]. AB - BACKGROUND: The amphiphilic interferon-inducing drug tilorone belongs to the group of synthetic fluorenone with antitumorus and antiviral effects and is known to cause lipidosis in the human and rat retina. METHODS: We treated female albino Wistar-Kyoto-Rats and Spontaneously-Hypertensive-Rats orally with tilorone for 8 weeks. The animals were submitted to electroretinography, and the retinae were prepared for histological investigations. RESULTS: Tilorone caused moderate lipidosis in the ganglion cells and slight lipidosis in the pigment epithelium after 8 weeks of treatment with a stronger effect in Spontaneously-Hypertensive Rats. The b-wave amplitude was reduced by 30% compared to controls in Spontaneously-Hypertensive-Rats and by 10% in the Wistar-Kyoto-group. The a-wave amplitude was reduced by 25% in Spontaneously-Hypertensive-Rats. CONCLUSION: Tilorone induced lipidosis shows a clear affinity to ganglion cells. The electroretinographic changes in Spontaneously-Hypertensive-Rats are most likely caused by the severe lipidosis in the neuroretina compared to the Wistar-Kyoto group. PMID- 10820708 TI - Intraocular concentrations of clindamycin obtained by sequential parabulbar injections. AB - PURPOSE: To measure the intraocular concentration of Clindamycin that can be maintained by sequential injection through a retained parabulbar catheter. MATERIALS AND METHODS: The peak intraocular concentration and half-life of Clindamycin after parabulbar injection was determined in a rabbit model. In a second experiment a parabulbar catheter was inserted and the maximum concentration that could be maintained by sequential injection through the catheter at intervals of 6 hours was determined. In a third experiment both eyes of the rabbit were catheterized and one infused with Clindamycin. The eyes were enucleated and studied by light and electron microscopy for changes induced by Clindamycin and the catheter. RESULTS: The concentration of Clindamycin after a single parabulbar injection of 10 mg/kg peaked in the retina and choroid at 2 hours and was 336 micrograms/g. The serum level at 2 hours was 2.2 micrograms/ml. The half-life in choroid and retina was 1 hour. Sequential administration at 6 hour intervals maintained a minimum concentration of 87 micrograms/ml. The vitreous concentration was maintained at 2 micrograms/ml. CONCLUSION: The level of Clindamycin in the retina and choroid obtained by sequential injections of 10 mg/kg exceeds the minimum lethal dose for organisms susceptible to the drug. The low serum concentration suggests that sequential doses of Clindamycin by the parabulbar route might be an effective therapy for toxoplasmic retinochoroiditis and would diminish the risk of colitis or other deleterious systemic side effects. PMID- 10820709 TI - [Bilateral inferior hemianopsia as an early symptom of Heidenhain type Creutzfeldt-Jakob disease]. AB - BACKGROUND: The Heidenhain variant is a subtype of Creutzfeldt-Jakob disease (CJD) where visual symptoms are leading. HISTORY AND SIGNS: We report on a 60 year-old woman who developed within a few weeks bilateral inferior hemianopsia. Morphological findings of the eyes were normal. Neurological examination and cerebral CT/MRI were normal too. P 100 latency of visual evoked potentials (VEP) of both eyes, however, was delayed. OUTCOME: During the course of the disease visual field narrowed progressively and 3 months after the first symptoms the patient went blind. Only at that time myoclonia, hyperkinesia, and psychiatric symptoms appeared. Changes of EEG and cerebrospinal fluid were suggestive of Creutzfeldt-Jakob disease (CJD). The patient died after 5 1/2 months and diagnosis was confirmed neuropathologically with mainly occipital spongiform encephalopathy. CONCLUSION: This case illustrates the diagnostic difficulties of the Heidenhain type of CJD and demonstrates that in patients with progressive visual field defects without morphological abnormalities of the eyes or radiological abnormalities of the brain CJD should be considered. It should be looked out for additional neurological (especially extrapyramidal) and psychiatric symptoms. PMID- 10820710 TI - [Foreign body granuloma after lower eyelid avulsion and surgical revision with silicone tube splint]. AB - BACKGROUND: Foreign-body-reactions with granulomas after use of silicone containing biomaterials are of clinical importance [4]. HISTORY AND SIGNS: A 4 year old patient entered our hospital with a reddish vascularised tumor. The tumor's origin was in the conjunctiva of the nasal lid-angle of the right eye. THERAPY AND OUTCOME: After a traumatic disconnection of the canaliculus lacrimalis inferior, a bicanalicular silicon-tube intubation had been performed with the patient in the age of one year. Only 3 years after this intervention a secondary clinical examination could be performed. At this time, an extended solid conjunctival-tumor appeared at the operated eye. After removal and histological examination it could be classified as a foreign body granuloma. CONCLUSIONS: The blood- and tissue compatibility of silicone is only insufficiently guaranteed on the basis of the hydrophobic material-surface. After reconstructive lacrimal surgery postoperative care and the removal of the stabilizing silicone material is necessary. PMID- 10820711 TI - [Walker-Warburg syndrome in adulthood?]. AB - BACKGROUND: Walker-Warburg-Syndrome (WWS) comprises malformations of the brain and the eyes with muscle dystrophy. Its course is usually lethal within the first 60 weeks of life. As far as we know there are no reports in the literature dealing with WWS in adults. CASE REPORT: A 70-year old female died because of cardio-pulmonary failure while she had an infection of the respiratory tract. The patient had been mentally deficient and had suffered from recurrent single convulsions. 17 years before death funduscopy had revealed an optic atrophy. Diabetes mellitus was known for ca. 15 years. The neuropathologists found a lissencephaly with a typical histology and a cerebellar malformation. Histologically, the eyes showed diabetic changes like vacuolization of the iris pigment epithelium and a thickening of basement membranes. There were marked deposits in the Pars plicata of the ciliary body and drusen formation in Bruch's membrane. The nerve fiber layer of the retina seemed to be thinned, and there was a considerable rarefication of optic ganglion cells. CONCLUSIONS: Taking the clinical and especially the morphological data into account, a Walker-Warburg Syndrome was assumed. Within the eyes developmental and diabetic changes were combined. Because the cerebral and ocular malformations were comparably mild, the patient became unusually old, and visual function allowed at least good orientation. References were searched by medline and by the indices of known papers. PMID- 10820712 TI - [20 MHz ultrasound diagnosis in chronic canaliculitis]. AB - BACKGROUND: High frequency sonography is used with a medium frequency of 50 to 100 MHz for non-invasive examination of the anterior eye segment. 20-MHz probes are considered to be a cost-efficient alternative, with a lower resolution and a higher penetration depth. We examined the value of the 20-MHz probe in diagnosing chronic canaliculitis. CASE REPORT: A 50-year-old woman was evaluated for a chronic canaliculitis. A pathognomonic sign of the chronic canaliculitis was the detection of concrements by high-resolution ultrasound. Grains measuring 1-2 mm in diameter could be shown in an ecstatic canaliculus with 20-MHz sonography. The high-resolution sonography showed the extent of the ectasia of the canaliculus and was therefore useful in planing the operative strategy. CONCLUSION: The spectrum of possibilities in diagnosing chronic canaliculitis is broadened with this diagnostic tool. Specially when characteristic symptoms are missing, the 20 MHz probe may be useful in diagnosing patients with chronic canaliculitis. PMID- 10820713 TI - Bacterial protein toxins. An overview. PMID- 10820714 TI - Purification of Clostridium botulinum type A neurotoxin. PMID- 10820715 TI - Shiga toxins. PMID- 10820716 TI - Isolation and detection of microcystins and nodularins, cyanobacterial peptide hepatotoxins. PMID- 10820717 TI - Genetic construction, expression, and characterization of diphtheria toxin-based growth factor fusion proteins. PMID- 10820718 TI - Structure-function analysis of cysteine-engineered entomopathogenic toxins. PMID- 10820719 TI - Use of Fourier-transformed infrared spectroscopy for secondary structure determination of staphylococcal pore-forming toxins. PMID- 10820720 TI - The use of fluorescence resonance energy transfer to detect conformational changes in protein toxins. PMID- 10820722 TI - Characterization of molecular properties of pore-forming toxins with planar lipid bilayers. PMID- 10820721 TI - Site-directed spin labeling of proteins. Applications to diphtheria toxin. PMID- 10820723 TI - Determination of affinity and kinetic rate constants using surface plasmon resonance. PMID- 10820724 TI - ADP-ribosylation of alpha-Gi proteins by pertussis toxin. Positional dissection of acceptor sites using membrane anchored synthetic peptides. PMID- 10820725 TI - Phage libraries for generation of anti-botulinum scFv antibodies. PMID- 10820726 TI - T-cell cytotoxicity assays for studying the functional interaction between the superantigen staphylococcal enterotoxin A and T-cell receptors. PMID- 10820727 TI - In vitro physiological studies on clostridial neurotoxins. Biological models and procedures for extracellular and intracellular application of toxins. PMID- 10820728 TI - The biology of endotoxin. PMID- 10820729 TI - Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry of lipopolysaccharides. PMID- 10820730 TI - Applications of combined capillary electrophoresis-electrospray mass spectrometry in the characterization of short-chain lipopolysaccharides. Haemophilus influenzae. PMID- 10820731 TI - Deacylation of lipopolysaccharides and isolation of oligosaccharide phosphates. PMID- 10820732 TI - Electrophysiological measurements on reconstituted outer membranes. PMID- 10820733 TI - Extraction and purification of an enzyme potentially involved in ABA biosynthesis. PMID- 10820734 TI - Differential display. Analysis of gene expression during plant development. PMID- 10820735 TI - Abscisic acid. ABA immunoassay and gas chromatography/mass spectrometry verification. PMID- 10820736 TI - Auxin analysis. PMID- 10820737 TI - Photoacoustic and photothermal detection of the plant hormone ethylene. PMID- 10820738 TI - Analysis of gibberellins. PMID- 10820739 TI - Cytokinins. Extraction, separation, and analysis. PMID- 10820740 TI - Binding studies. PMID- 10820741 TI - Mutagenesis. PMID- 10820742 TI - The identification of ethene biosynthetic genes by gene silencing. Antisense transgenes, agrobacterium-mediated transformation, and the tomato ACC oxidase cDNA. PMID- 10820743 TI - Extraction, separation, and analysis of plant phosphoinositides and complex glycolipids. PMID- 10820744 TI - Reverse genetics. Screening plant populations for gene knockouts. PMID- 10820745 TI - Isolation of large gel-forming mucins. PMID- 10820746 TI - Preparation of membrane mucin. PMID- 10820747 TI - Histologically based methods for detection of mucin. PMID- 10820748 TI - Detection and quantitation of mucins using chemical, lectin, and antibody methods. PMID- 10820749 TI - The gastrointestinal adherent mucous gel barrier. PMID- 10820750 TI - Quantitation of biosynthesis and secretion of mucin using metabolic labeling. PMID- 10820751 TI - Separation and identification of mucins and their glycoforms. PMID- 10820752 TI - Heterogeneity and size distribution of gel-forming mucins. PMID- 10820753 TI - Rheology of mucin. PMID- 10820754 TI - Amino acid analysis of mucins. PMID- 10820755 TI - Identification of glycosylation sites in mucin peptides by edman degradation. PMID- 10820756 TI - Synthetic peptides for the analysis and preparation of antimucin antibodies. PMID- 10820757 TI - Mucin domains to explore disulfide-dependent dimer formation. PMID- 10820758 TI - Monosaccharide composition of mucins. PMID- 10820759 TI - O-linked oligosaccharide chain release and fractionation. PMID- 10820760 TI - Structural analysis of mucin-type O-linked oligosaccharides. PMID- 10820761 TI - Measurement of sulfate in mucins. PMID- 10820762 TI - Biosynthesis of mucin cell and organ culture methods for biosynthetic study. PMID- 10820763 TI - Metabolic labeling methods for the preparation and biosynthetic study of mucin. PMID- 10820764 TI - Identification of mucins using metabolic labeling, immunoprecipitation, and gel electrophoresis. PMID- 10820765 TI - Mucin precursors. Identification and analysis of their intracellular processing. PMID- 10820766 TI - Inhibition of mucin glycosylation. PMID- 10820767 TI - O-linked chain glycosyltransferases. PMID- 10820768 TI - Mucin cDNA cloning. PMID- 10820769 TI - Northern blot analysis of large mRNAs. PMID- 10820770 TI - Southern blot analysis of large DNA fragments. PMID- 10820771 TI - In situ hybridization techniques for localizing mucin mRNA. PMID- 10820772 TI - Detection of mucin gene polymorphism. PMID- 10820773 TI - Polyclonal and monoclonal techniques. PMID- 10820774 TI - Monoclonal antibodies to mucin VNTR peptides. PMID- 10820775 TI - Mucinase activity. PMID- 10820776 TI - Proteinase activity. PMID- 10820777 TI - Glycosidase activity. PMID- 10820778 TI - Assays for bacterial mucin-desulfating sulfatases. PMID- 10820780 TI - Growth of mucin degrading bacteria in biofilms. PMID- 10820779 TI - Mucin-bacterial binding assays. PMID- 10820781 TI - Generation of MUC1 cytotoxic T-cells in mice and epitope mapping. PMID- 10820782 TI - Analysis of the frequency of MHC-unrestricted MUC1-specific cytotoxic T-cells in peripheral blood by limiting dilution assay. PMID- 10820783 TI - Expression of MUC1 in insect cells using recombinant baculovirus. PMID- 10820784 TI - Mucin antigen presentation using dendritic cells. PMID- 10820785 TI - Detection of humoral immune responses to mucins. PMID- 10820786 TI - Peniamidienone and penidilamine, plant growth regulators produced by the fungus Penicillium sp. No. 13. AB - Peniamidienone and penidilamine were isolated from cultures of the fungus Penicillium sp. No. 13 as new plant growth regulators and their structures were established by NMR spectroscopic studies. Peniamidienone showed weak inhibition of lettuce seedling growth. PMID- 10820787 TI - Anti-platelet aggregation constituents from Gynura elliptica. AB - A p-hydroxyacetophenone-like derivative, (+)-gynunone, and a chromane, together with six known compounds were isolated from the CHCl3 fraction of the roots of Gynura elliptica. Their structures were determined by means of spectral analyses. Among the isolates, 6-acetyl-2,2-dimethylchroman-4-one and vanillin showed anti platelet aggregation activity induced by arachidonic acid in vitro. PMID- 10820788 TI - Iridoids from Dunnia sinensis. AB - A plumieride type iridoid glucoside, dunnisinoside, and a non-glucosidic iridoid, dunnisinin, were isolated from the leaves of Dunnia sinensis. Their structures were established by 1D and 2D NMR and FABMS experiments. PMID- 10820789 TI - Cassane diterpenoid from Caesalpinia major. AB - A new cassane diterpenoid, 14-deoxy-epsilon-caesalpin was isolated from the seed kernels of Caesalpinia major and its structure was determined by spectroscopic data. PMID- 10820790 TI - Sesquiterpenoids and diterpenoids from the Chilean liverwort Lepicolea ochroleuca. AB - The ether extract of the Chilean liverwort Lepicolea ochroleuca yielded three sesquiterpenoids, ent-4 beta-Hydroxy-10 alpha-methoxyaromadendrane, ent-3 beta Hydroxyspathulenol, and 1,10-Dioxotayloriane, as minor components. The major components were ledol and 13-epi-neoverrucosan-5 beta-ol, four other minor fusicoccanoids were identified. PMID- 10820791 TI - Synthesis of methyl dihydrohardwickiate and its C-4 epimer. Structural amendment of natural crolechinic acid. AB - Reduction of the alpha, beta-unsaturated ester moiety of (+)-methyl hardwickiate with magnesium in methanol afforded methyl (4aS,6S,8aS,1R,5R)-5, 6,8a-trimethyl-5 [2'-(3"-oxoyl)-ethyl-perhydro-1-naphthalenyl]- carboxaylate, while reduction with sodium in n-propanol, followed by esterification with diazomethane, furnished its C-4 epimer. After comparison of the 1H- and 13C-NMR data of these compounds with those reported for crolechinic acid isolated from Croton lechleri, a stereochemical revision for the natural product is suggested. PMID- 10820792 TI - ent-Kaurene diterpenoids from Isodon rubescens. AB - Two kaurenoids, taibairubescensins A and B, were isolated from the ethanol extract of the leaves and branches of Isodon rubescens. Their structures are designated as 2 beta, 3 beta-diacetoxy-11 beta, 13 alpha-dihydroxy-ent-kaur-16-en 15-one and 3 beta, 11 beta-diacetoxy-2 beta, 6 alpha-dihydroxy-ent-kaur-16-en-15 one, respectively, on the basis of detailed spectroscopic analyses. PMID- 10820793 TI - Triterpenoid saponins containing an acetylated branched D-fucosyl residue from Quillaja saponaria Molina. AB - Seven novel saponins were isolated from a bark extract of Quillaja saponaria Molina. the compounds were characterized, using mainly NMR spectroscopy, mass spectrometry and chemical methods, as quillaic acid substituted at C-3 with oligosaccharides consisting of various compositions of D-glucuronic acid D galactose, D-xylose, and L-rhamnose and at C-28 with complex oligosaccharide structures consisting of various compositions of D-xylose, L-rhamnose, D-apiose and a branched 4-O-acetyl-D-fucose residue. PMID- 10820794 TI - Cyclopentene dialdehydes from Tabebuia impetiginosa. AB - The isolation of two cyclopentene dialdehydes, 2-formyl-5-(4'-methoxybenzoyloxy) 3-methyl-2-cyclopentene-1-acetal dehyde, and 2-formyl-5-(3', 4' dimethoxybenzoyloxy)-3-methyl-2-cyclopentene-1-acetaldehyde, from the bark of Tabebuia impetiginosa is reported. The structures were established by analysis of spectroscopic data. These compounds showed anti-inflammatory activity. PMID- 10820795 TI - A glycosidic eudesmanolide from Hyaloseris salicifolia. AB - The flowers of Hyaloseris salicifolia afforded known ivasperin, while the leaves afforded ivasperin and 2-O-(6'-O-acetyl-beta-D-glucopyranosyl)-ivasperin, whose structure was determined by spectroscopic methods. PMID- 10820796 TI - A cytotoxic diterpene from Alomia myriadenia. AB - An extract of the aerial parts from Alomia myriadenia Schultz-Bip. ex Baker (Asteraceae) showed significant cytotoxicity against a panel of human cancer cell lines in a screening of extracts from Brazilian Atlantic Forest plant species. Employing a bioassay-linked HPLC-electrospray/MS method, followed by semi preparative HPLC, the active component was isolated and characterized as a mixture of epimers of the labdane diterpene 12S,16-dihydroxy-ent-labda-7,13-dien 15,16-olide. PMID- 10820797 TI - Sesterterpenoid from Gentianella alborosea. AB - The structure of a new type of sesterterpenoid, designated as alborosin, isolated from Gentianella alborosea, has been deduced from a spectroscopic investigation. PMID- 10820798 TI - Triterpenoid saponins and sapogenin lactones from Albizia gummifera. AB - The structures of two new monodesmosidic and bisdesmosidic triterpenoid saponins (1 and 2) and the known compound delta 5-stigmasterol-3-O-beta-D-glucopyranoside (3) as well as two new oleanane type triterpene lactone glycosides 4, 5 and a new sapogenin lactone 6 isolated from the stem bark of Albizia gummifera C.A. Smith (Mimosaceae) have been elucidated as 3-O-?beta-D-glucopyranosyl(1-->2)-[alpha-L arabinopyranosyl(1-->6) ]-beta-D- glucopyranosyl?-oleanolic acid (1), beta-D glucopyranosyl(1-->2)-beta-D-glucopyranosyl 3-O-?beta-D-glucopyra-nosyl(1-->2) [alpha-L-arabinopyrano syl(1-->6)]-beta-D- glucopyranosyl?-oleanolate (2), 3 beta ?O-D-glucopyranosyl-(1-->2)-[O-alpha-L-arabinopyranosyl(1-->6 )] beta-D glucopyranosyloxy?-machaerinic acid gamma-lactone (4), 3 beta-O-beta-D glucopyranosiduronic acid (1-->2)-beta-D-glucopyranosyloxy]-machaerinic acid gamma-lactone (5), and A-homo-3a-oxa-5 beta-olean-12-en-3-one-28-oic acid (6), respectively. The complete assignment of the 1H and 13C resonances of 1, 2, 4 and 6 and of the peracetate of 5 were achieved by means of 2D-NMR studies. PMID- 10820799 TI - Oleanane saponins from Gymnema sylvestre. AB - Six oleanane-type saponins, along with two known triterpene saponins, were isolated from the leaves of Gymnema sylvestre. The structures of the oleanane triterpene glycosides were characterized as longispinogenin 3-O-beta-D glucuronopyranoside, 21 beta-benzoylsitakisogenin 3-O-beta-D-glucuronopyranoside, 3-O-beta-D-glucopyranosyl(1-->6)-beta-D-glucopyranosyl oleanolic acid 28-O-beta-D glucopyranosyl ester, oleanolic acid 3-O-beta-D-xylopyranosyl(1-->6)-beta-D glucopyranosyl(1-->6)-beta-D- glucopyranoside, 3-O-beta-D-xylopyranosyl(1-->6) beta-D-glucopyranosyl (1-->6)-beta-D-glucopyranosyl oleanolic acid 28-O-beta-D glucopyranosyl ester and 3-O-beta-D-glucopyranosyl(1-->6)-beta-D-glucopyranosyl oleanolic acid 28-beta-D-glucopyranosyl(1-->6)-beta-D-glucopyranosyl ester on the basis of hydrolysis and spectral evidence, including 1D- and 2D-NMR (TOCSY, ROESY, HMQC and HMBC) and FABMS analyses. PMID- 10820800 TI - Triterpenoid glycosides and a triterpene from Ilex brevicuspis. AB - Two saponins and a sapogenin were isolated from the leaves of Ilex brevicuspis. Their structures were established by means of spectroscopic methods as brevicuspisaponin 1 (3-O-alpha-L-arabinopyranosyl-20(S)-19 alpha,24 dihydroxyursolic acid), brevicuspisaponin 2 (3-O-alpha-L-arabinopyranosyl-20(S) 19 alpha,23,24-trihydroxyursolic acid) and 23-methylester of 20(S)-3 beta,19 alpha,24-trihydroxyurs-12-en-23,28-dioic acid. PMID- 10820801 TI - Triterpene saponins from the fruits of Hedera helix. AB - Six triterpene saponins, including two new compounds, were isolated from the fruits of Hedera helix L. (Araliaceae). The structures of the new compounds, named helixosides A and B, were established as 3-O-beta-D-glucopyranosyl-(1-->2) beta-D-glucopyranosyl hederagenin 28-O-beta-D-glucopyranosyl-(1-->6)-beta-D glucopyranosyl ester, and 3-O-beta-D-glucopyranosyl-(1-->2)-beta-D-glucopyranosyl oleanolic acid 28-O-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl ester, respectively, on the basis of chemical and spectral data. PMID- 10820802 TI - Icetexane and abietane diterpenoids from Salvia gilliessi. AB - One icetexane and two abietane diterpenes were isolated from the aerial parts of Salvia gilliesii, and characterized as 5-epi-icetexone; 12-hydroxy-11,14-diketo 6,8,12-abietatrien-19,20-olide and 6 alpha,12,19-trihydroxy-11,14-diketo-8,12 abietadien-20,7 beta-olide, respectively. The structures were established by analysis of their 1H and 13C NMR spectra with the aid of 2D experiments. The triterpene oleanolic acid was isolated from the same source. PMID- 10820803 TI - A cleistanthane diterpene lactone from Vellozia compacta. AB - 8,11,13-cleistanthatrien-7-one-19,20 beta-olide and six other previously described diterpenes were isolated by silica gel chromatography of the ethyl alcohol extract of Vellozia compacta. The structure of the cleistanthane lactone was deduced on the basis of spectral data analysis. PMID- 10820804 TI - Highly oxygenated ecdysteroids from Vitex canescens root bark. AB - Highly oxygenated ecdysteroids, (24R)-11 alpha, 20,24-trihydroxyecdysone and 11 alpha,20,26-trihydroxyecdysone, have been isolated from the polar fraction of Vitex canescens root bark. The latter exists as two C-25 epimers which could be separated by reversed-phase HPLC. PMID- 10820805 TI - Iridals from Iris tectorum and Belamcanda chinensis. AB - Three iridals, iridotectorals A and B, and iridobelamal. A, were isolated from rhizomes of Iris tectorum and Belamcanda chinensis, respectively, along with five known iridals. Their structures were elucidated on the basis of spectral evidence. The human promyelocytic leukemia (HL-60) cell-adhesion activity of the eight iridals is also discussed. PMID- 10820806 TI - Phlorigidosides A-C, iridoid glucosides from Phlomis rigida. AB - From the aerial parts of Phlomis rigida, three iridoid glucosides, phlorigidoside A (2-O-acetyllamiridoside), B (8-O-acetyl-6-beta-hydroxyipolamide) and C (5 deoxysesamoside), were isolated together with the known iridoid glucosides, shanzhiside methyl ester, 8-O-acetylshanzhiside methyl ester, deoxypulcheloside I, lamiridoside, and 6-beta-hydroxyipolamide. The structures of the new compounds were elucidated based on spectral and chemical evidence. PMID- 10820807 TI - Monoterpene and pregnane glucosides from Solenostemma argel. AB - From the aerial parts of Solenostemma argel, two monoterpene glucosides have been isolated and identified as 6,7-dihydroxy-dihydrolinalool 3-O-beta-glucopyranoside and 6,7-dihydroxy-dihydrolinalool 7-O-beta-glucopyranoside. A pregnane glucoside was also isolated and assigned as pregn-5-ene-3,14-beta-dihydroxy-7,20-dione 3-O beta-glucopyranoside together with the known compounds benzyl alcohol O-beta apiofuranosyl-(1-->6)-beta-glucopyranoside, 2-phenylethyl O-alpha arabinopyranosyl-(1-->6)-beta-glucopyranoside, astragalin and kaempferol-3-O alpha-rhamnopyranosyl-(1-->2)-beta-glucopyranoside. PMID- 10820808 TI - Triterpenoid saponins from the roots of tea plant (Camellia sinensis var. assamica). AB - Three olean-12-ene type triterpenoid saponins, named TR-saponins A, B and C, were isolated as methyl esters from tea roots (Camellia sinesis var. assamica) after treatment with diazomethane. Their structures were established as the methyl esters of 3-O-alpha-L-arabinopyranosyl (1-->3)-beta-D-glucuronopyranosyl-21, 22 di-O-angeloyl-R1-barrigenol-23-oic acid, 3-O-alpha-L-arabinopyranosyl (1-->3) beta-D-glucuronopyranosyl-21-O-angeloyl-22-O-2-me thylbutanoyl-R1- barrigenol-23 oic acid and 3-O-alpha-L-arabinopyranosyl (1-->3)-beta-D-glucuronopyranosyl-16 alpha-O-acetyl-21-O-angeloyl-22-O-2-methylbutanoyl-R1-bar rigenol-23-oic acid, by extensive 1D and 2D-NMR as well as FABMS and HR-MS analyses. PMID- 10820809 TI - Constituents and bioactivity of the tubers of Euphorbia sessiliflora. AB - The diterpene ent-12-hydroxy-12[R]-abieta-8(14),13(15)-dien-16,12-olide was isolated from the tubers of Euphorbia sessiliflora Roxb., together with four known ent-abietadienolides, four known cycloartane triterpenes and ellagic acid beta-D-glucopyranoside. Two of these metabolates displayed moderate antibacterial activities. PMID- 10820810 TI - Diterpenoids from Salvia miltiorrhiza. AB - The abietane diterpenoid, neocryptotanshinone II, and the known 6,12 dihydroxyabieta-5,8,11,13-tetraen-7-one were isolated as minor components from the roots of Salvia miltiorrhiza. Their structures were established on the basis of spectral evidence. PMID- 10820811 TI - Diterpenoids from Humirianthera ampla. AB - Two diterpenoids, humirianthol and acrenol, as well as the known annonalide, were isolated from Humirianthera ampla. Humirianthol and acrenol were determined by 1D and 2D NMR spectroscopic techniques to be 3 beta,20:14 beta,16-diepoxy-3 alpha, 15 alpha-dihydroxy-7-pimaren-19,6 beta-olide and 3 beta,20-epoxy-3 alpha,15,16 trihydroxy-7-pimaren-19,6 beta-olide, respectively. PMID- 10820812 TI - Mulinane-type diterpenoids from Laretia acaulis. AB - In addition to the known mulinolic acid and mulin-11, 13-dien-20-oic acid, 13 epimulinolic acid has been isolated from the aerial parts of Laretia acaulis (Cav.) Gill et Hook (Umbelliferae). Its structure was based on spectroscopic comparison with mulinolic acid and by chemical characterization. PMID- 10820813 TI - 6-Hydroxyluteolin-7-O-(1''-alpha-rhamnoside) from Vriesea sanguinolenta Cogn. and Marchal (Bromeliaceae). AB - The isolation of 6-hydroxyluteolin-7-O-(1"-alpha-rhamnoside) from the Central American epiphyte Vriesea sanguinolenta Cogn. and Marchal (Bromeliaceae) is described here. Its stereostructure was established by spectroscopic methods and an X-ray structure analysis of its hepta-O-acetyl derivative. This flavonoid glycoside had previously been reported from some Teucrium species (Labiatae), yet without sufficient physical data and spectroscopic evidence. PMID- 10820814 TI - Minor diarylheptanoid glycosides of Alnus rubra bark. AB - The diarylheptanoid (S)-1,7-bis-(4-hydroxyphenyl)-heptan-3-one-5-O-beta-D xylopyranosi de, and two known compounds, 1,7-bis-(3,4-dihydroxyphenyl)-heptan-3 one-5-O-beta-D-glucopyranos ide and platyphylloside were isolated from Alnus rubra bark. Structures were established by application of spectrometric techniques. PMID- 10820815 TI - Two chromene derivatives from Calyptranthes tricona. AB - Two alpha-monomethyl chromene derivatives were isolated from the leaf essential oil of Calyptranthes tricona from Brazil which were characterized by 1H- and 13C NMR. Besides these components, which represent about half of the oil, classical terpenoid structures could be identified, among which cis-beta-farnesene is the most abundant (26.6%). A biosynthetic pathway could be proposed to explain the formation of the chromene derivatives in the plant. PMID- 10820816 TI - Indicanines B and C, two isoflavonoid derivatives from the root bark of Erythrina indica. AB - In addition to two known compounds, 5,4'-di-O-methylalpinumisoflavone and cajanin, a new 3-phenylcoumarin metabolite, named indicanine B, and a new isoflavone derivative, named indicanine C, were isolated from the root bark of Erythrina indica. By means of spectroscopic analysis, the structures of the new compounds were characterized as 4-hydroxy-3-(4'-hydroxyphenyl)-5-methoxy-2",2" dimethylpyrano [5",6":6,7] coumarin and 4'-hydroxy-5-methoxy-2",2"-dimethylpyrano [5",6":6,7] isoflavone, respectively. The 13C-NMR data of cajanin and the in vitro antimicrobial spectrum and potencies of the isolated compounds are also reported. PMID- 10820817 TI - Phenanthrenes from Dendrobium plicatile. AB - From Dendrobium plicatile stems, three phenanthrenes were isolated. The structures are 2,5-dihydroxy-4,9,10-trimethoxyphenanthrene, 2,5-dihydroxy-4 methoxyphenanthrene and 2,5,9-trihydroxy-4-methoxy-9,10-dihydrophenanthrene. PMID- 10820818 TI - Chalconoids from Fissistigma bracteolatum. AB - Phytochemical studies on the leaves of Fissistigma bracteolatum yielded besides the two known compounds 2-hydroxy-3,4,6-trimethoxychalcone (1) and 5,7,8 trimethoxyflav-3-ene (2), five new chalconoids 2-hydroxy-3,4,6-trimethoxychalcene (3), 2-hydroxy-3,4,6-trimethoxydihydrochalcone (4), 2'-hydroxy-3',4',6' trimethoxydihydrochalcone (5), 2'-hydroxy-3',4',6'-trimethoxy-beta' methoxychalcane (6) and 2'-hydroxy-3',4',6'-trimethoxy-beta'-ethoxychalcane (7). The structures of these compounds were determined by mass and NMR spectroscopic methods. PMID- 10820819 TI - Flavonoids from Goodyera schlechtendaliana. AB - A flavonol glycoside, 3-[[6-O-(6-deoxy-alpha-L-mannopyranosyl)-beta-D- glucopyranosyl]oxi]-5,7-dihydroxy-8-[(4-hydroxy-3,5-dimethoxyphenyl)meth yl]- 2 (3,4-dihydroxypheny)-4H-1-benzopyran-4-one, trivially named goodyerin, was isolated from the whole plant of Goodyera schlechtendaliana, along with three known flavonoids, rutin, kaempferol-3-O-rutinoside and isorhamnetin-3-O rutinoside. The structures were established by spectroscopic analysis. PMID- 10820820 TI - Two glycosides from the stem bark of Tetracentron sinense. AB - Two glycosides, tetracentronsides A and B, were isolated from the stem bark of Tetracentron sinense Oliv., along with ten known compounds, beta-sitosterol, lupeol, betulinic acid, oleanolic acid, vanillic aldehyde, vanillic acid, maslinic acid, huazhongilexin, daucosterol and catechin. On the basis of spectral and chemical evidence, tetracentronside A and B were identified as 3,4,5 trimethoxyphenyl-O-6'-O-vanilloyl-beta-D-glucopyranoside and (8R, 8'R) 9-beta-D glucopyranosyl dihydrocubebin, respectively. PMID- 10820821 TI - A bichalcone from the twigs of Rhus pyroides. AB - The twigs of Rhus pyroides yielded a novel bichalcone 2',4",2"'-trihydroxy-4',4"' dimethoxy-4-O-5"'-bichalcone. It was identified on the basis of spectroscopic data including 1D and 2D NMR spectroscopy. The name rhuschalcone-1 is proposed. PMID- 10820822 TI - Stilbene oligomers in roots of Sophora davidii. AB - Three stilbene oligomers, davidiols A-C were isolated from the roots of Sophora davidii in addition to the seven known phenols, leachianone A, sophoraflavanones G, H and I, miyabenol C, alpha-viniferin and epsilon-viniferin. Their structures and relative configurations were established by means of 2D-NMR spectroscopy including COLOC and PSNOESY. PMID- 10820823 TI - Stilbenoids in the stem bark of Hopea parviflora. AB - From the bark of Hopea parviflora, two stilbenoids, named (+)-parviflorol and (-) ampelopsin A, were isolated in addition to three known compounds: (+) balanocarpol, (-)-epsilon-viniferin and (-)-hopeaphenol. Their structures were determined by analysis of spectral data, including 2D NMR and NOE experiments, respectively. PMID- 10820824 TI - Xanthones from Calophyllum teysmannii var. inophylloide. AB - Further study of the wood of Calophyllum teysmannii Miq. var. inophylloide from Thailand yielded the xanthones 1,2,8-trimethoxyxanthone, 6-hydroxy-1,2,5,7 tetramethoxyxanthone and 3-(3'-carboxybutyl)-4-methoxyxanthone (teysmannic acid) in addition to scriblitifolic acid, 1,7-dihydroxyxanthone and the benzoic acid derivative leiocarpic acid. PMID- 10820825 TI - Coumarins from Ferulago capillaris and F. brachyloba. AB - Four new coumarins, (+)-senecioylprangol, (-)-3'-senecioyloxymarmesin, (+)-3' hydroxyprantschimgin and (+)-2"-senecioyloxymarmesin, besides 12 known coumarins have been isolated from two Ferulago species. Their structures have been established by spectroscopic methods and partial synthesis. New synthetic 3' oxocoumarins are also described. There is a remarkable difference in the contents of the most abundant coumarins found in the roots of both species: osthol and aurapten are specific to F. capillaris and F. brachyloba, respectively. PMID- 10820826 TI - 7-Caffeoylsedoheptulose from Nyssa sylvatica. AB - 7-Caffeoylsedoheptulose has been isolated from the wood of Nyssa sylvatica, along with two ellagic acid derivatives and scopoletin. Its structure was established by chemical and spectral evidence. PMID- 10820827 TI - Structure elucidation of clavilactone D: an inhibitor of protein tyrosine kinases. AB - Clavilactones D and E were isolated from an agar culture of the Basidiomycetous fungus Clitocybe clavipes, and their structure was elucidated by 1H- and 13C-NMR studies. Clavilactone D is an inhibitor of tyrosine kinases. PMID- 10820828 TI - Xanthone and dihydroisocoumarin from Montrouziera sphaeroidea. AB - A xanthone, montrouxanthone and a dihydroisocoumarin, montroumarin were isolated from the stem bark of Montrouziera sphaeroidea Pancher Ex Planchon et Triana [Guttiferae], along with two known compounds. Their structures were elucidated on the basis of spectroscopic analyses. This is the first report of the analysis of chemical constituents of Montrouziera species. PMID- 10820829 TI - Flavonoids from Brosimum acutifolium. AB - 4'-Hydroxy-7,8-[2-(2-hydroxyisopropyl)dihydrofuran]flavan and 4',7-dihydroxy-8 (3,3-dimethylallyl)flavan, together with 10 known plant constituents, were obtained from the trunk bark of Brosimum acutifolium. Their structures were elucidated by spectroscopic methods, including 2D NMR spectroscopic techniques. PMID- 10820830 TI - Phenylpropanoid glycosides from Smilax glabra. AB - Five phenylpropanoid esters of sucrose glycosides, trivially named smiglasides A E, were isolated from the rhizomes of Smilax glabra. Their structures were elucidated on the basis of spectroscopic studies. PMID- 10820831 TI - Quaternary indole alkaloids from the stem bark of Strychnos guianensis. AB - Two new quaternary alkaloids, 9-methoxy-Nb-methylgeissoschizol and guiachrysine together with the known compounds C-alkaloid O, fluorocurine, mavacurine, macusine B and C-profluorocurine, were isolated from Strychnos guianensis stembark. The structures of the compounds were elucidated on the basis of spectroscopic studies. PMID- 10820832 TI - Aporphinoid alkaloids and other constituents from Lettowianthus stellatus. AB - Two new aporphinoid alkaloids, f1ttowianthine and 11-methoxylettowianthine were isolated from the root bark of Lettowianthus stellatus, together with the new sesquiterpene 11-hydroxyguaia-4,6-diene and the known compounds liriodenine, (Z) 7-octadecen-9-ynoic acid, methyl (2E,6E,10R)-10,11-epoxy-3,7,11-trimethyl-2,6 dodecadienoate, methyl (2E,6E,10R)-10,11-dihydroxy-3,7,11-trimethyl-2,6 dodecadienoate , and 3,4,5-trimethoxyphenol. The structure elucidation was achieved by spectroscopic methods. PMID- 10820833 TI - Alkaloids and phenylpropanoids from Peganum nigellastrum. AB - Two canthin-6-one alkaloids, luotonins C and D, and two phenylpropanoids, dihydrosinapyl ferulate and dihydroconiferyl ferulate, were isolated from the aerial parts of Peganum nigellastrum along with four known alkaloids, harmine, 3 phenylquinoline, 3-(4-hydroxyphenyl)quinoline and 3-(1H-indol-3-yl)quinoline. Their structures were elucidated by spectroscopic techniques. The structures of luotonins C and D were also confirmed by chemical synthesis. PMID- 10820834 TI - An azaanthracene alkaloid from Polyalthia suberosa. AB - An azaanthracene alkaloid, 1-aza-9,10-dimethoxy-4-methyl-2-oxo-1,2 dihydroanthracene (kalasinamide) has been isolated from the stems of Polyalthia suberosa. In addition, the known N-trans-feruloyltyramine and N-trans coumaroyltyramine are also reported from the same source. The structures were elucidated by spectroscopic methods. PMID- 10820835 TI - Volatile constituents in juice and oil of Australian wild lime (Microcitrus inodora). AB - Fifty-three volatile constituents from the juice and twenty from the peel oil of Microcitrus inodora have been identified by gas chromatographic and mass spectral analysis. All except seven had been reported earlier as citrus constituents. Since M. inodora is used as a parent for production of new citrus hybrids, this information will be useful to horticulturists, plant breeders and phytochemists. PMID- 10820837 TI - Three putrescine bisamides from the leaves of Aglaia grandis. AB - Three putrescine (i.e. 1,4-butanediamine) bisamides were isolated from the leaves of Aglaia grandis. Their structures were elucidated by interpretation of spectral data. PMID- 10820836 TI - Structure and absolute configuration of new acetylenic compounds isolated from cultures of Clitocybe catinus. AB - Investigations of the extracts of a culture of Clitocybe catinus gave rise to the isolation of new acetylenic diols 1-3. Their structure was determined on the basis of 1H- and 13C-NMR evidence and the absolute configuration elucidated by means of the modified Mosher's method. PMID- 10820838 TI - Procyanidins from the seeds of Vitis amurensis. AB - Three procyanidins, procyanidin B-5 3'-O-gallate, vitisinol, and amurensisin, were isolated from the seeds of Vitis amurensis, whose structures were elucidated on the basis of spectral and chemical evidence. Vitisinol and amurensisin contain a spiro-type biflavanyl linkage and a biphenyl-lactone partial structure, respectively. PMID- 10820839 TI - Nor-lignan and sesquiterpenes from Cremanthodium ellisii. AB - A nor-lignan and two sesquiterpenes, along with six known compounds, have been isolated from the medicinal plant Cremanthodium ellisii. Their structures were determined on the basis of spectral evidence, especially 2D NMR (1H-1H COSY, HMQC, HMBC). PMID- 10820840 TI - The prototropic tautomerism of nitrogen bases: a new insight into the old problem. AB - The summary is made of the series of theoretical calculations performed by the semiempirical quantum-chemical AM1 method of physico-chemical features of prototropic tautomerism of nucleotide bases with full optimization of all parameters. Applying of nontraditional approach based on the view that the highest possible number of prototropic tautomers of bases are determined by an overall quantity of protons and electron lone pairs of proton acceptor atoms, the success was achieved in ascertaining that, as regards prototropic tautomerism, nucleotide bases are more mobile structures (prototropic tautomerism of nucleotide bases has a molecular-zwitterionic character involving carboprotons as well) than it was assumed earlier. A biological significance of the data obtained, which are generalized and may be extended from the authors point of view to more wide range of objects, amino acids included. PMID- 10820841 TI - [Membrane mechanisms of regulating Ca ion concentration in smooth muscle cells. II. System of passive Ca2+ transport in smooth muscles]. AB - The aim of this review is to summarize some current aspects on the membrane mechanisms of energy-independent Ca2+ transport in the smooth muscles. The emphasis is placed on the characteristics of voltage-gated and receptor-operated calcium channels of plasma membrane and two major Ca2+ release channels of sarco(endo)plasmic reticulum: one is activated by IP3 and sensitive to heparin and the other by Ca2+ and sensitive to ryanodine. A brief discussion of the electro- and pharmacomechanical coupling mechanisms in the smooth muscle is given. PMID- 10820842 TI - [Effect of vitamin D3 on metabolic processes in rat erythrocytes under extreme factors]. AB - The effects of vitamin D3 (as an ingredient of the preparation "Videchol") on the activity of some glycolysis enzymes (lactic dehydrogenase, LDH; pyruvate kinase PK; hexokinase HK) and of pentose phosphate shunt (glucose-6-phosphate dehydrogenase), and also on the changes of metabolites concentration levels: ATP, NAD+, pyruvate and lactate following the administration of the vitamin to the control group rats and animals irradiated by low rate ionizing radiation were established. "Videchol" administration leads to the of LDH activity decrease within 10 days, of HK activity within 20 days and to the decrease of PK activity within 5 and 20 days after the experiment beginning. In the low dose radiation conditions a reconnection of carbohydrates' metabolism from glycolisys into pentose phosphate pathway in the erythrocytes was observed. The administration of vitamin D3 as a component of the preparation "Videchol" following the low rate radiation background facilitates the normalization of LDG, PK and HK activities in comparison with the same parameters of only irradiated rats. The studies on LDH--reaction kinetics showed the redistribution of the LDH isozymes pattern with different K(m) means for pyruvate in the rats subjected to the joint action of low dose chronical radiation and "Videchol" occurred. PMID- 10820843 TI - [pH-dependence of kinetic parameters in the superprecipitation reaction and ATPase activity of myometrial actomyosin]. AB - The investigation of pH-dependence of superprecipitation reaction and ATPase activity of myometrium actomyosin in the interval of pH 5.5-8.0 has detected cupola-shaped curves with maximal activity of both processes by pH 6.5. On the basis of calculating the constants of ionization it was supposed that in the case of actomyosin ATPase imidazole groups of two histidins had an essential role in reaction of ATP hydrolysis and in superprecipitation process--imidazol group of histidine and carboxyl group of asparagin acid. The investigation of [ATP]- and [Mg2+]-dependence of superprecipitation reaction by pH 6.0, 6.5 and 7.0 has demonstrated different pH-sensitiveness of Michaelis constants and maximal speeds relatively Mg2+ and ATP for both processes. It was shown that pH-optimum of ATPase activity of myometrium actomyosin coincided with maximal affinity of actomyosin with ATP and Mg2+ while as for superprecipitation reaction the correlation between value of process by certain pH and affinity with ATP and Mg2+ was not detected. PMID- 10820844 TI - [Effect of melatonin and epithalamin on activity of marker enzymes in the cell membrane in the forebrain of rats under acute hypoxia]. AB - The effects of a single-shot intraperitoneally administration of melatonin in a dose of 1 mg per kg body weight and epithalamin in a dose of 2.5 mg per kg body weight on the activities of Na+, K(+)-ATPase and 5'-nucleotidase were investigated in the forebrain of juvenile male white rats under the acute hypobaric hypoxia. The melatonin and epithalamin administration against the background of acute hypoxia prevented an acute hypoxia inducing decrease in the activity of Na+, K(+)-ATPase as well as increased in the activity of 5' nucleotidase. Such effects of pineal hormones can promote antihypoxic protection of neurons. PMID- 10820845 TI - [Characteristics of antioxidant properties of 20-hydroxyecdysone in low density lipoproteins by kinetic parameters of chemiluminescence]. AB - 20-Hydroxyecdysone is capable to terminate the lipid free-radical oxidation in low density lipoprotein in vitro as displayed by the kinetic chemiluminescence parameters. In concentrations interval from 2 x 10(-6) mol/l up to 8 x 10(-6) mol/l it statistically reliably reduces maximum of the first flash intensity of the low density lipoprotein Fe(2+)-initiated chemiluminescence. It testifies about Fe(2+)-initiated lipid peroxide process decomposition in low density lipoprotein slowing down by 20-hydroxyecdysone. In concentration of 20 hydroxyecdysone 8 x 10(-6) mol/l the statistically reliable reduction of angle tangent of an ascending branch of the second flash of low density lipoprotein Fe(2+)-initiated chemiluminescence was found. This is a result of free-radical lipid oxidation rate reduction in the low density lipoprotein at the 20 hydroxyeodysone presence. The equations of the kinetic parameters linear dependences Fe(2+)-initiated chemiluminescence in low density lipoprotein on the concentration of 20-hydroxyeodysone and hydroquinone have been received. The correlation factors in the interval from 0.8980 up to 0.6789 have been calculated. Thus, the assumption has been forwarded that 20-hydroxyeodysone has antioxidizing properties. However, its antioxidizing activity in free radical lipid oxidation of is low density lipoprotein is as less as for hydroquinone. PMID- 10820846 TI - [Effect of amizon on structural modifications of liver nuclear chromatin from rats, intoxicated with tetrachloromethane]. AB - The actions of the amizon on the antioxidant activity and free radicaloformation level in the nuclear chromatin fractions of rat liver methan tetrachlorid intoxication was studied using the Fe(+2)-induction biochemiluminescence and microcalorimetry methods. The amizon genomoprotective effect on the chromatin transcriptionally active fraction was shown. PMID- 10820847 TI - [Isolation and characteristics of antistreptokinase antibodies from blood serum of myocardial infarct patients treated with streptokinase]. AB - Antistreptokinase IgG (antiSK IgG) from blood of 8 patients with acute myocardial infarction that were treated by streptokinase (SK) has been investigated. AntiSK IgG contained 1.8% of total serum IgG. They had high affinity to SK (K50% approximately 10 nM) and inhibited activation of plasminogen (Pg) by SK with K50% approximately 6 nM. AntiSK IgG were bound with chymothriptic fragments of SK with affinity decreased in the set of fragments: 47 > 36 > 30 > 17 > or = 11 > 7 kDa. 11 linear epitopes of antiSK IgG were localized in I1-S12, T43-M70, G139-Q152, T163-I190, T193-S222, F241-Y252, Y275-P286, T315-L336, I365-E376, S379-T390 and Y397-N410 sites of SK primary structure using SK decapeptides. 70% of antibodies were bound with T43-M70 (38.3%), T315-L336 (13.2%) and Y397-N410 (17.7%) SK sequences located in alpha and gamma SK domains. By depletion of antiSK IgG on Pg SK complex it was shown that 80-85% of antiSK IgG bound to Pg-SK complex, 47.9% of that contained antibodies with epitopes located in I1-S12, T43-M70, T193-S222 and S379-T390 SK sequences, and rest of IgG had probably spatial epitopes. Unbound with Pg-SK complex antibodies inhibited activation of Pg by SK with higher affinity (Ki approximately 1.2 nM) in comparison with total antiSK IgG fraction. The role of different sites in antigenity of SK and in Pg-SK complex formation is discussed. PMID- 10820848 TI - Partial dependence of human peripheral blood leukocyte binding to high molecular weight fucoidan on divalent cations. AB - L-selectin (CD62L) is the principal leukocyte adhesion molecule for the high endothelial venules of peripheral lymph nodes. This adhesion has an absolute requirement for calcium ions. Nevertheless, some studies have shown carbohydrate adhesion receptor interactions on lymphocytes and neutrophils, including the L selectin molecule, that are Ca-independent. In the present study fucoidan, a reportedly Ca2+ independent ligand of L-selectin, and Mabs to human CD62L were coupled to magnetic polystyrene beads (MPB), as a model of leukocyte-surface interactions, and the efficiency of human leukocyte separation was investigated. 30% of Ficoll-purified human mononuclear cells and 75% of dextran-purified human leukocytes (DPHL) were specifically bound by fucoidan-modified MPB in the presence of Ca2+; 55% of dextran-purified leukocytes were specifically bound in the absence of Ca2+. The specific binding was inhibited by an excess of free fucoidan. The data obtained show the presence of Ca-independent adhesion determinants, specific to fucoidan on human leukocytes. No significant specific binding of leukocytes to fucoidan-modified MPB was found after the incubation with fresh human Ca(2+)-depleted whole blood. More than 90% of DPHL were specifically bound to MPB modified with Mabs to human CD62L irrespective of Ca2+ presence. The same degree of separation was achieved after the incubation with fresh human Ca(2+)-depleted-whole blood with anti-CD62L modified beads. PMID- 10820849 TI - [Oxidation of low density lipoproteins in the presence of silica gel treated with fullerene]. AB - This study estimated the effects of low density lipoprotein (LDL) oxidation in the presence of fullerene silicagel in comparison with ions of Cu+2 in the wide region of oxidation times and reagent concentrations. Investigation is directed to reveal the reagent parameters, that promote to increase the oxidation rate of the LDL solution. It revealed only qualitative coincidence of the LDL oxidation kinetics in the presence as Cu+2 as fullerene, that indicated on the identity of the oxidation mechanism in both cases. At the same time the revealed quantitative difference between the oxidation process parameters are interpreted in the terms of oxidation potential and the electron affinity that characterize the reagent activity in the reaction accompanied by the electron transfer. The good correlation between above mentioned parameters and the oxidation rate of the aqueous LDL solution is observed: its increasing takes place at the increase of the electron affinity or at the decrease of the oxidation potential. PMID- 10820850 TI - [Functional status of cryopreserved rat liver mitochondria]. AB - Mitochondria from the rat liver have been frozen down to -196 degrees C under protection of dimethylsulfoxide, after which they were subjected to periodical increase and decrease in temperature (cycling) in the ranges of -105 degrees C to -196 degrees C or -135 degrees to -196 degrees C. There have been observed some non-lethal damages of mitochondria which were revealed in the decrease in the respiratory control and in the ADP/O ratio after the cycling. The damage rate increased with the increase of the number of temperature decrease-increase cycles. The damages were also greater in the case when the upper limit of cycling temperature was higher than the glass transition temperature of the freezing medium (Tg = -126 degrees C). The non-lethal damages of mitochondria are explained by the influence of electric fields, appearing in the frozen sample during the temperature increase or decrease. PMID- 10820851 TI - [Compensation for changes in the acid-base balance in intact newborn calves and in experimental metabolic acidosis and alkalosis]. AB - The quantitative parameters of the new-born calves acid-alkali balance in the early period of their life has been defined to be essentially changed. Considering the latter their organism acid-base balance natural stabilization is characterized by the certain dynamics which is being disturbed displays a quick development of adaptive reactions. The new-born ones' organism capability to support the acid-base homeostasis has been identified as considerably determined by their tissues alkaline reserve level. PMID- 10820852 TI - [Status of the rat antioxidant system and correction by medicinal plant preparations during the dynamics of growth of Heren's carcinoma]. AB - The work deals with the experimental researches of lipids peroxidation in the rats liver and reduced glutatione and some antioxidant enzymes activity in the same animals blood and liver in dependence on the Heren's carcinoma dynamics as well as in the tumor-carriers ones injected subcutaneously by "NIKA" herbal preparation. As the result of the experiments it was identified, that in the course of the tumor growing there was an increase of the rats liver level of malonic dialdehyde--the final product of the lipids peroxidation, as well as of some antioxidant enzymes activity growth in the blood and liver. In the 21-th day following the tumor implantation the animals-tumor-carriers antioxidant system is exhausted. The tested herbal medicine displays the normalizing and correcting action on the rats organism protective system and provides for the tumor growth slowering. PMID- 10820853 TI - [Metabolism of heme and hemoproteins in rat liver upon administration of mercuric chloride]. AB - Rat liver delta-aminolevulinate synthase (delta-ALAS) activity in the early period after mercury chloride administration (0.7 mg per 100 g body weight) was found to be followed by free heme level increase, which was registered by the increase of heme saturation of the heme-binding protein tryptophan-2,3 dioxygenase (T-2,3-DO). delta-ALAS and heme oxygenase activity increase was observed 24 h after action. Microsomal cytochromes P450 and b5 levels decrease. Heme saturation of the T-2,3-DO returned to control level. Heme oxygenase and T 2,3-DO induction promoted hepatocytes free heme level normalization. Heme oxygenase and delta-ALAS induction role in the liver cells defense from the oxidative damage is discussed. PMID- 10820855 TI - [Determination of antibody affinity using ELISA]. AB - Some new approaches for the determination of antibody affinity were proposed. It was pointed out that the proposed methods are more simple, convenient, precise, and informative than that of Friguet et al. (1985). The approach that allows determination of two-valence antibodies affinity was also proposed. The example of two monovalent antibodies presented in the examined mixture was considered. It allows to estimate the affinity of both kinds of antibodies as well as to determine their concentration relations in the mixture. PMID- 10820854 TI - [Effect of eosin Y on Ca2+-dependent activity of Ca2+-Mg2+-ATPase in smooth muscle cell membrane]. AB - With the aim to elucidate mechanism of eosin Y inhibitory effect on the Ca(2+) transporting ATPase activity of myometrial cell plasma membrane effect of this inhibitor on the maximal initial rate of ATP hydrolysis reaction, catalyzed by Ca2+, Mg(2+)-ATPase, and on the enzyme affinity for Ca2+ was studied. It was established that eosin Y decreased the rate of Ca2+, Mg(2+)-ATPase catalitic turnover determined by Ca2+ and had no effect on enzyme affinity for this cation. PMID- 10820856 TI - [A quantitative method for determining lectin activity on microplates]. AB - A modification of a known quantitative solidphase microtiter plate lectin assay elaborated by Hatakeyama and coauthors developed with biotinylated lectins and ExtrAvidin--alkaline phosphatase conjugate was proposed. The modification was performed in order to use a broad spectra of lectin ligands, including glycopeptides, peptides, etc. The optimal concentrations of the reagents and time of the reagents incubation were selected. Being known from the literature Kdiss for Concanavalin A and monosaccharides: methyl-Man, Man, Glc and Gal were determined in a model experiments. The results were in agreement with the range of monosaccharides specificity for the Concanavalin A. The modification described is relatively simple and sensitive. PMID- 10820857 TI - [A convenient method for laboratory synthesis of the N-([1 14C]palmitoyl)ethanolamine, labeled by a fatty acid]. AB - Data concerning the synthesis of bioactive lipid compound N-([1-14C] palmitoyl)ethanolamine labeled by 14C fatty acid are reported. The method is based on the ability of ethanolamine and fatty acid to the direct chemical condensation at 180 degrees C with yielding of N-acylethanolamine. The purification of the end product by the double crystallization in ethanol allows to obtain chromatographically pure substance. The presented method of the labeled N-([1-14C]-palmitoyl)ethanolamine synthesis is simple and in extensive that's why it might be used in the area of biologically active compounds investigation. PMID- 10820858 TI - [Structural regularities in activated cleavage sites of thrombin receptors]. AB - Comparison of thrombin receptors activation splitting sites sequences testifies to their similarity both in activation splitting sites of protein precursors and protein proteinase inhibitors reactive sites. In all these sites corresponded to effectory sites P2'-positions are placed by hydrophobic amino-acids only. The regularity defined conforms with previous thesis about the role of effectory S2' site in regulation of the processes mediated by serine proteinases. PMID- 10820859 TI - [Validation of the new German translation version of the "Short Inflammatory Bowel Disease Questionnaire" (SIBDQ)]. AB - In the face of a hardly predictable course of illness--potentially effecting a severe diminution of daily activities--the quality of life (QOL) represents a chief outcome measure in the treatment of inflammatory bowel diseases (IBD). For the German-speaking area an economical disease-specific instrument measuring QOL has been missing, so far. Therefore, we translated and adapted the recently validated "Short Inflammatory Bowel Disease Questionnaire" SIBDQ (11). Bowel symptoms, systemic symptoms, moods, and social activities are represented by a total of 10 items. Along with this questionnaire 7 accepted instruments assessing QOL-components were administered to 125 outpatients (54 ulcerative colitis, 71 Crohn's disease) and 51 healthy controls. The psychometric properties are good. Consistency is alpha = .84, thereby slightly surpassing the Anglo-American SIBDQ (alpha = .78). Retest-reliability after 26 weeks is r = .60. Systematic covariations with accepted QOL-instruments give evidence for its convergent and discriminatory validity. Differences between groups (high vs. low inflammatory activity vs. healthy controls) substantiate its sensitivity. Thus, the German adaptation of the accepted SIBDQ supplies an economic, reliable, and valid instrument that proved able to assess clinically relevant differences in QOL in patients with IBD. We suggest that it should be used by its overall score rather than its subscales. The instrument is appropriate to be used in therapy outcome and evaluation studies. It allows international comparison. PMID- 10820860 TI - [Budesonide foam as a new therapeutic principle in distal ulcerative colitis in comparison with mesalazine enema. An open, controlled, randomized and prospective multicenter pilot study]. AB - Budesonide is a locally acting steroid with a high first-pass metabolism in the liver and low systemic effects. We performed the first pilot study comparing budesonide foam (1 mg/50 ml b.i.d.) with mesalazine enemas (4 g/60 ml o.d.). 33 patients from 3 centres were enrolled in this open randomized clinical trial. 16 patients got budesonide foam and 17 got mesalazine enemas. The drugs were administered for 4 weeks. Histological index (HI) and endoscopic index (EI) were assessed at day 1 and day 28, clinical activity index (CAI) at day 1, 14 and 28. For patients with left-sided colitis and proctosigmoiditis improvement was defined as decrease in CAI of > or = 2 points. For patients with proctitis improvement was defined as decrease in HI of > or = 1 point. The primary efficacy evaluation was performed with the intention to treat population (n = 32). Improvement was found in 67% of the patients in the budesonide group compared to 71% in the mesalazine group. There was no statistically significant difference between the groups. Adverse events were mild and rare in both groups. Both treatments had no significant influence on plasma cortisol. In this pilot study for distal ulcerative colitis budesonide foam is as safe and effective as mesalazine enema. PMID- 10820861 TI - Sonographic measurement of thickened bowel wall segments as a quantitative parameter for activity in inflammatory bowel disease. AB - Inflammatory bowel disease (IBD) is associated with morphological changes of the bowel wall that can be visualized by abdominal ultrasound (US). This method is a tool to detect the extent of bowel wall thickening and the length of involved segments. The purpose of this study was to determine the value of sonographic measurement of inflamed bowel wall segments as a quantitative parameter for disease activity. 137 patients with Crohn's disease (CD) and 32 patients with ulcerative colitis (UC) were included in the present study. A total 356 US examinations were performed within a one-year period. In a segment-by-segment analysis we determined the "volume of inflamed bowel wall" (VIB) by measuring wall thickness and longitudinal extent of pathologically altered bowel segments. VIB was used as a quantitative parameter for disease activity based on sonomorphological findings. At the same time the following parameters were also determined: CD activity index (CDAI) in patients with CD, clinical activity index (CAI) in patients with UC, erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP). We found no relevant correlation between VIB and biochemical indices of inflammation (ESR, CRP) and between VIB and clinical activity of IBD (CDAI, CAI). All correlation coefficients were below 0.5. It can be concluded that the extent of inflammatory changes of the bowel wall detected by US is not strictly associated with clinical activity and laboratory parameters of inflammation. PMID- 10820862 TI - [Obstructive jaundice caused by spontaneous rupture of an Echinococcus granulosus cyst into the bile duct system]. AB - Obstructive jaundice caused by a parasitosis is very rare. It can be a complication of a cystic echinococcosis. We present the case of a 33-year-old man who suffered from cystic echinococcosis with an affection of the liver for several years. As a complication an echinococcal cyst had ruptured into the biliary tract and had led to an occlusion of the ductus hepatocholedochus. By means of ERCP the membrane of the echinococcal liver cyst was extracted in toto. Cholangioscopy followed and showed a free biliary tract without remaining cyst fragments. After these interventions the blood parameters normalized and the patient recovered. An additive chemotherapy with albendazole was started. PMID- 10820863 TI - Campylobacter jejuni-induced severe colitis--a rare cause of toxic megacolon. AB - The development of toxic megacolon as a sequel of infectious colitis is rare. We have observed the very rare case of a campylobacter jejuni-induced toxic megacolon. A 28-year-old man was admitted with severe enterocolitis and appearance of blood in stools. He had been treated with loperamide without success. Two days after admission stool cultures revealed campylobacter jejuni and then an oral antibiotic therapy was started. On the fifth day clinical performance deteriorated again with development of toxic megacolon and consecutive subtotal colectomy. Rectoscopy before discharge after 13 days showed a normal mucosa. The unusual course with first improvement and then rapid deterioration despite adequate therapy was observed in 4 other cases, which may also be a hint of ensuing megacolon. Even in usually harmless enterocolitis like campylobacter infection, predisposing factors such as loperamide are known to precipitate toxic megacolon and should be considered in clinical practice. PMID- 10820864 TI - Malignant peritoneal mesothelioma with mimicry of pseudomyxoma peritonei in a patient with a history of perforated sigmadiverticulitis. AB - We describe a 57-year-old man who presented with diffuse abdominal pain, abdominal enlargement, vomitus, dyspnea and a weight loss of 30 kg within 6 months. These acute symptoms were preceded by an episode of ascites and an acute sigmadiverticulitis 7 months ago. Ultrasonography and computed tomography were suggestive of pseudomyxoma peritonei. However, malignant mesothelioma peritonei was diagnosed by open surgery with biopsy for histological examination. Despite R 2-resection of the tumor and following open hyperthermic intraperitoneal chemotherapy with initial remarkable recovery the patient died 5 months after therapeutical intervention. Malignant peritoneal mesothelioma is an extremely rare tumor with great diagnostic and therapeutic difficulties. We report a case including diagnostical work up and the medical surgical therapy of this disease. PMID- 10820865 TI - [Helicobacter pylori and coronary heart diseases--hypotheses and facts]. AB - In 1994 Mendall et al. (9) have suggested that there might be a correlation between Helicobacter pylori (HP) infection and coronary heart disease (CHD), mediated by a chronic low-grade acute phase reaction with mildly raised serum or plasma concentrations of C-reactive protein and fibrinogen. According to this hypothesis, a gastric HP colonization might be an additional risk factor for CHD. In the meantime, 35 studies have examined whether HP seropositivity is associated with CHD occurrence. However, in 8 publications only CHD was definitively proven (in CHD+ patients) or excluded (in the corresponding control groups) by coronary artery angiography, and in only 2 of them (1 abstract, 1 full-length publication) a significant association between HP positivity (serologically proven) and CHD was ascertained. Additionally, a metaanalysis of 18 studies including 10,000 patients could not demonstrate any correlations between HP seropositivity and different acute phase proteins (66). Thus, a positive correlation between gastric HP colonization and CHD is far from being proven. Further proposed links between HP infection and CHD such as hyperhomocysteinemia (67) or autoimmune mechanisms (71) due to cross-reacting antibodies to HP HSP60/65 (heat shock protein) with human endothel-derived HSP60/65 need further confirmation. PMID- 10820866 TI - [Results of the expert workshop "Enteral Nutrition Therapy" of the German Society of Nutrition Medicine, Bonn, 14 June 1999. Key points]. PMID- 10820867 TI - [Regional chemotherapy of the liver after resection of liver metastases in colorectal carcinoma]. PMID- 10820868 TI - [Mutations of the lamin A/C gene causing familial partial lipodystrophy: significance for the development of hyperlipidemia and insulin resistant diabetes mellitus]. PMID- 10820869 TI - [How likely is sexual transmission of hepatitis C viruses?]. PMID- 10820870 TI - [Changes in gait caused by plateau shoes]. PMID- 10820871 TI - [Value of synovectomy in surgical rheumatology]. PMID- 10820872 TI - [Design of a multicenter study for assessing the effectiveness of extracorporeal shockwave therapy in epicondylitis humeri radialis]. AB - INTRODUCTION: Previously published studies concerning, extracorporeal shock-wave therapy (ESWT) in the treatment of lateral epicondylitis do not fulfil the biometric standards of modern clinical research. The objective of the trial is to show that ESWT is effective in the treatment of chronic LE. METHOD: A prospective, randomized, placebo-controlled, single-blinded, multicenter trial with an independent blinded observer was designed. The effectiveness of ESWT is evaluated by comparison with a control group in which sham-ESWT is performed, both under local anaesthesia. Outcome is determined on the basis of the Roles/Maudsley-Score. Inclusion criteria are a history of at least 6 months of LE and failure of conventional treatment. The therapy includes 3 sessions of low energy ESWT with 2000 impulses (energy flux density 0.07-0.09 mJ/mm2). Sample size is 272 patients. STATUS: Randomisation started in October 1998 and is planned over a period of two and a half years. CONCLUSION: Only a randomised clinical trial with adequate control of placebo effects and observer bias can provide the required evidence for the efficiency of ESWT in the treatment of lateral epicondylitis of the elbow. PMID- 10820873 TI - [Revisions after arthroscopic interventions in the subacromial space]. AB - PROBLEM: In the present study, we evaluated the failure and revisions rates after arthroscopic subacromial decompression (SAD). METHOD: We examined 83 patients who were treated with arthroscopic subacromial decompression for primary shoulder impingement (stage II and stage III according to Neer) at an average follow-up time of 30 months. Hereby, special attention was paid to the revision operations resulting from our treatment. RESULTS: In patients with impingement stage II, the mean follow-up Constant-score was 84.7 (SD +/- 16.7) while in patients with impingement stage III it was 78.0 (SD +/- 21.8). In ten patients (12%) revision operation had to be performed, nine of them with initial stage II impingement and one with initial stage III impingement. Reasons for revisions were persisting or increasing pain as well as functional dissatisfaction. At the follow up examination, six of these revised patients (60%) were satisfied with the result. The mean follow up Constant-score in the revised patients was 77.3 (SD +/- 17.4). In 60% of the revised patients the necessity for the revision operation was directly related to the technical problems at the primary operation, in 40% we found reasons not related to the primary SAD. CONCLUSIONS: Technical falls are the most common cause for the need of revision operations after SAD. This demonstrates how demanding this kind of operation procedure is. In case of revision operations individual strategies should be developed in which the decision whether to perform arthroscopic or open revision procedures is of special importance. PMID- 10820874 TI - [Relevant differences after post-traumatic and degenerative humeral head replacement (intermediate-term results)]. AB - INTRODUCTION: In a comparative study, mid-term findings after posttraumatic and degenerative hemi-prosthetic shoulder joint replacement were analyzed and compared with current reports in the literature. METHODS: The patient sample in the retrospective study consisted of 20 patients with proximal fractures of the humeral head (x = 64 years) and 6 patients (x = 61 years) with degenerative shoulder joint disease. In 14 patients the humeral head was replaced as a primary procedure at < 4 weeks after the trauma and in 6 patients as a secondary treatment at > 4 weeks. The follow-up period was 38-41 months on average. Evaluation was based on the Constant Score, subjective assessment by the patient, and conventional radiographs. RESULTS: After posttraumatic shoulder replacement, the patients achieved a Constant Score of 65 (+/- 19) and after degenerative replacement 74 (+/- 34) points. In the posttraumatic group, the Constant Score after primary implantation was better than after secondary implantation (68 +/- 16 versus 59 +/- 15 points). After posttraumatic replacement, the only improvement over time was in pain levels (p < 0.05). 92% of all patients were satisfied with the result and would accept the same treatment again. Radiologically, a clinical correlate could be found for the arrosions at the acromion and occasionally for the secondary upwards displacement of the humerus. The results correlated well with data from the literature. CONCLUSIONS: For posttraumatic conditions, hemiarthroplasty led to better results within four weeks and seemed to be a suitable alternative to other procedures, especially in older patients. Patients with primary osteoarthritis and idiopathic humeral head necrosis can expect good to excellent results after hemiarthroplasty. In both groups, the overall results depend mainly upon patient compliance and the state of the rotator cuff. PMID- 10820875 TI - [Results of surgical therapy of mobility impaired elbow joints]. AB - AIM OF THE STUDY: Analysis of results after operative treatment of elbow stiffness in a retrospective clinical study. METHODS: 36 patients were evaluated by an independent observer with a mean follow-up of 53 months (7-128). The mean age at operation was 32.9 years (4-64). Post-traumatic origins of elbow stiffness were differentiated from non-traumatic origins. The evaluation was done according to a modified Morrey-Score with 37 points of range of motion, 30 points of pain and 12 points of activity of daily living. The absolute and relative improvement of range of motion was calculated. The strength of elbow flexion and extension was measured with a isocinetic dynamometer in 23 patients. RESULTS: The mean score improved from 46.6 to 67.4 points postoperative (p < 0.001). Range of motion improved from 30.1 to 34.2, pain from 9.9 to 23.1 and ADL from 6.6 to 10.1 points. The mean relative improvement of range of motion for flexion-extension was 49%, for pronation-supination 25%. The strength of flexion was 78.5%, the strength of extension 77.4% of the unaffected contralateral side. The results were affected by grade, duration and origin of elbow stiffness. CONCLUSION: The operative therapy of post-traumatic and non-traumatic elbow stiffness is an effective option of treatment, when conservative therapy failed. The range of motion improved, and the patients were able to manage 80% of the activities of daily living. PMID- 10820876 TI - [Decompensation of a supinator syndrome in submuscular lipoma. A rare differential diagnosis of elbow pain]. AB - Important differential diagnoses to epicondylitis humeri radialis are the nerve compression syndromes of the elbow. After a long period of conservative treatment, paresis of the motor branch of nervus radialis in this case led to the diagnosis and surgical treatment showing an unknown submuscular lipoma as the cause of a supinator syndrome with paresis of the finger extensors. In cases of therapy-resistant pain of the elbow, especially resistant to the conservative therapy of an epicondylitis humeri radialis, a nontraumatic supinator syndrome should be considered as a differential diagnosis. A tumor as the cause of a compression syndrome of the motor branch of the nervus radialis is very rare. PMID- 10820877 TI - [Differential diagnostic considerations in microcephalic dwarfism]. AB - PURPOSE: While the rare Seckel-Syndrome is defined by clear criteria, clinical and radiologic findings for microcephalic osteodysplastic primordial dwarfism (MOPD) make an exact diagnosis and classification difficult. By comparing our patients to previously described cases of MOPD we evaluate the hypothesis that this disorder has a greater heterogeneity than has been believed up until now. Furthermore the differential diagnosis of the MOPD-complex is discussed. RESULTS: Two cases that show typical growth retardation, microcephalus and facial anomalies as well as osteodysplastic deformities including hip dysplasia are presented. The parents of both children are consanguineous and of Arabic race. In one of the children growth hormone levels were noticeably decreased. In discrepancy to the Seckel-syndrome both children showed no signs of mental retardation, therefore the classification into the heterogeneous group of microcephalic osteodysplastic primordial dwarfism (MOPD) is the most likely diagnosis. CONCLUSION: It is suggested that microcephalic osteodysplastic primordial dwarfism (MOPD) has a greater clinical and radiological expression than has been assumed up until now. Whether our results are merely a variant or suggest a new subtype of the MOPD can only be resolved by further cases. The exact pathogenesis of the disease currently remains unknown but the most probable cause is an autosomal recessive inheritance. PMID- 10820878 TI - [Scoliosis and hip flexion contracture in Duchenne muscular dystrophy]. AB - Spinal deformity is common in muscular dystrophy and usually occurs after loss of walking ability. Unlike in idiopathic and other scoliosis forms, there seems to be no side preference of the convexity. Aim of the study was to analyse, if there is any relation between incidence and extent of walking ability, lower limb contractures and development of scoliosis. METHODS: In a retrospective study, 45 patients with Duchenne muscular dystrophy who underwent surgery were analysed, concerning walking ability, contractures of lower extremities and scoliosis. RESULTS: 1: No scoliosis was observed in ambulatory patients. 2: 96% of the wheelchair bound patients suffered from scoliosis. 3: 96% of the scoliosis patients had hip flexion or abduction contractures. 4: In 12 of 15 cases with side-different contractures, scoliosis tended to the side with the greater contracture. CONCLUSIONS: The influence of hip contracture and pelvic obliquity on scoliosis is discussed controversially. Concerning muscular dystrophy, there seems to be a positive correlation between convexity and hip contracture. If this is a causal relation or if there is a faster progression of structural alteration of the muscles on one side has to be further investigated. PMID- 10820879 TI - [Intraoperative hyperpyrexia in patients with osteogenesis imperfecta]. AB - The aim of this study was to evaluate the influence of different types of anesthesia upon the intra- and postoperative body temperature in osteogenesis imperfecta patients. The development of an intraoperative hyperpyrexia of unknown origin is a typical phenomenon in patients with osteogenesis imperfecta. Body temperatures of up to 40 degrees C are known to complicate the operation. Therefore, in a retrospective study, the pre-, intra- and postoperative body temperature curves of 45 operations under different anaesthesias were measured. Group A underwent a common balanced anaesthesia with the volatile anaesthetic Enfluran in combination with Fentanyl, while group B was operated on under total intravenous anaesthesia (TIVA) with Propofol and Alfentanil. The preoperative temperatures were not different in the two groups. The intraoperative curves showed a constant body core temperature or even an increase under Enfluran, while the temperature always decreased under TIVA. This could be confirmed by intraindividual studies in 5 patients. PMID- 10820881 TI - [Anti-osteolytic therapy preserves trabecular structure and mechanical properties of bone in tumor osteolysis]. AB - PURPOSE OF THE STUDY: Little is known about the effect of a tumor on the trabecular architecture, therefore we employed an animal model for the assessment of bone quality in tumor osteolysis to determine the alterations of the trabecular architecture in tumor osteolysis and after an interventional treatment with a bisphosphonate. METHODS: To assess the bone mass and the micro architecture of the trabecular bone in tumor osteolysis we employed a micro computed tomography system. For the assessment of the mechanical properties of the treated and non-treated tumor-bearing bones we used a torsion test. RESULTS: The presence of a tumor in bone resulted in a reduction of bone mass, stability and architectural parameters. An interventional treatment of the animals with a bisphosphonate increased the bone mineral content, mechanical and architectural parameters compared to the non-treated, tumor-bearing animals. CONCLUSIONS: These results clearly show a beneficial effect of an anti-osteolytic treatment with a bisphosphonate in regard of bone quality in tumor-induced osteolysis. PMID- 10820880 TI - [Minimal invasive therapy of localized Langerhans-cell histiocytosis of bone]. AB - PURPOSE: Scaglietti introduced a method of steroid injection for the management of unicameral bone cysts in 1974. Subsequently the intralesional infiltration of corticosteriods has also been recommended as a primary therapy for localized Langerhanscell histiocytosis (LCH). We report our experience with the administration of methylprednisolone acetate in children and young adults localized LCH. MATERIAL AND METHODS: [corrected] Nine patients with localized LCH, aged 2 3/12 to 29 years, were treated with a single--in only one case two- intralesional injection of methylprednisolone acetate as a crystalline suspension. The dose was between 40 and 150 mg depending on the size of the radiolucent defect. We treated 4 lesions in the skull, 3 in the femur, 1 in the distal humerus and 1 in the mandibula. In each case the diagnosis was established by biopsy. Follow-up ranged from 2 to 8 8/12 years (4 years 4 months on average). RESULTS: 7 out of 9 patients with localized LCH had excellent results with complete healing of the lesion. In 2 patients there was no response to the initial injection therapy and dissemination of the disease occurred. In 4 patients with an additional soft tissue tumor, after injection therapy of the bone lesion, the soft tissue tumor resolved without further treatment. CONCLUSION: Intralesional infiltration of methylprednisolone acetate as a primary therapy for localized Langerhans cell histiocytosis leads to rapid relief of pain, restoration of bone morphology and reduction of associated soft tissue tumors. Performed with appropriate skill under sterile condition with the reported high percentage of effectiveness and low recurrence rate, this low invasive method is the treatment of choice, resulting in a lower morbidity and lower costs. PMID- 10820883 TI - [2 benign bone lesions in a tubular bone: misinterpretation of clinical and radiological findings]. AB - INTRODUCTION: Bone lesions are common at puberty. Most of them are detected by chance at examination for other reasons. The nonossifying fibroma does not produce any clinical symptoms and normally is diagnosed by chance. A spontaneous healing is discussed. The osteoid osteoma is characterized by its nocturnal pain and relief of pain at therapy with salicylates. Operative procedures are indicated for healing. CASE: A 12-year-old girl presented with radiological signs of a nonossifying fibroma. Because of local radionuclid uptake and clinical symptoms an operative procedure was carried out. Postoperative remaining pain resulted in radiological detection of a nearby osteoid osteoma and local operative therapy. CONCLUSION: Two benign bone lesions at one tubular bone are an extremely rare situation. The clinical symptoms and the radionuclid uptake are misinterpretated and falsely assigned to the nonossifying fibroma. All clinical symptoms and radiological signs have to be correlate correctly with the diagnosis and consecutive therapy in order to avoid a misinterpretation of obvious clear radiological results. PMID- 10820882 TI - [Surgical reconstruction of fibrous dysplasia of bone in long-term follow-up]. AB - PURPOSE: To examine how different operative measures influence the surgical outcome in patients with fibrous dysplasia of bone. METHODS: 118 dysplastic fibrous lesions of bone were surgically treated and reviewed in 70 patients between 1983 to 1993 (eleven years) with a median follow-up of six and a half years. Surgery consisted of intralesional curettage in 93 and marginal en bloc resection in 25 lesions. Bony defects were reconstructed with autogenous iliac crest graft in 55 lesions, with autogenous fibula graft in 9, with homologous bone chips in 28, and 5 times with a homologous fibula graft from the bone bank. In 33 lesions the entire defect was filled with polymethylmethacrylate. Osteosynthesis was performed in 41 patients. RESULTS: Recurrences requiring surgical revision were observed in 26 of 74 primary lesions (= 35% overall recurrence rate) at a mean 123.6 weeks postoperatively. The most frequent primary and recurrence location was the proximal femur (85% revision rate). 69% of all recurrences occurred under the age of 20. After intralesional curettage the reoperation rate was 32% and after marginal resection 8%. After reconstruction with autogenous iliac crest graft recurrence rate was 36%, after autogenous fibula graft 55%, after homologous bone chips 18%, after polymethylmethacrylate 9% and allograft fibula reconstruction showed no recurrences. A combined stable osteosynthesis bridging the fibrous osseous defect significantly reduced the revision rate to 3% (p = 0.01). CONCLUSION: Intralesional curettage and reconstruction with autogenous iliac crest graft in fibrous dysplasia of bone leads to a high recurrence rate. Reconstruction with cortical grafts or bone chips from the bone bank, if necessary in combination with a durable osteosynthesis in mechanically demanding locations, or solely bone cement in mechanically less demanding areas, reduces the revision rate in patients with monoostotic and polyostotic fibrous dysplasia. PMID- 10820884 TI - [Dorsal lumbar interbody implantation of cages for stabilizing segmental spinal instabilities]. AB - PURPOSE: The technique of posterior lumbar interbody fusion (PLIF) has been critically discussed due to a high degree of complications, including the development of pseudarthrosis. With the recent establishment of intercorporeal implants new aspects have to be contemplated in surgical techniques, especially concerning the posterior approach. In this study we present our first results after intercorporeal stabilisation of segmental spinal instabilities utilising carbon and titanium cages. METHOD: 45 spinal instabilities were surgically stabilised in 42 patients who were evaluated on average for 2.8 years post operatively. 12 patients had isthmic and 19 patients degenerative instabilities while 11 patients suffered from instabilities resulting from prior spinal surgery. RESULTS: Assessed according to the Hambly-score, 69% of the patients had an excellent or good result; 2 (4.8%) patients were subjectively worse off than before surgical treatment. After implantation of cages precise radiological evaluation of bony ingrowth is frequently impaired by artefacts. We found that three months after implantation of a titanium cage, which had to be removed after incorrect placement, no bony consolidation was visible. Persisting or recurrent instabilities in fused segments were not recorded. CONCLUSION: By means of PLIF and implantation of cages the interbody space is reconstructed and jeopardized neural structures are decompressed. In addition to this, the frequently osteochondrotically degenerated segment is immobilized. The posterior approach allows decompression of neural structures and, with comparable results concerning stability, the considerable risks of the ventral approach are avoided. PMID- 10820885 TI - [Implantation of a Harms titanium mesh cylinder for vertebral body replacement in spinal metastases]. AB - OBJECTIVE: Vertebral metastases are quiet common and may result in vertebral collapse, spinal instability, and progressive neurological compromise. An anterior approach is the accepted method for decompression and stabilization by vertebral replacement. There is still need for discussion regarding the material of the replacement device. Here, the authors report on Harms' titanium cage (DePuy-Motech) for vertebral replacement. METHODS: From 1992 until 1995 17 patients with spinal metastases were treated by anterior vertebral replacement at the Orthopaedic Department, University of Magdeburg. Indications for the procedure were single metastases and respectively, progressive neurological deficit with vertebral destruction seen radiologically. Implantation of the cage was performed solely or in combination with anterior or posterior instrumentation. Pre- and postoperative assessments were evaluated by clinical scores. RESULTS: Mean follow-up was 9.3 months. At the end of our study 11 patients had died (mean survival 6.7 months). Mean follow-up of the surviving 6 was 9.8 months. Patient's daily activity (Karnofsky) improved in 8 cases, became worse in 4 cases, and did not change in 5 cases. Neurologically (Frankel) there was improvement in 3 patients, deterioration in 4 patients, and no change in 10 patients. Walking ability showed improvement in 6 cases, deterioration in 3 cases and no change in 8 cases. Relief of pain (Moskowitz) was reported by 12 patients, no patient complained of increasing pain, 5 patients reported no change, and 10 patients were pain-free. CONCLUSION AND CLINICAL RELEVANCE: In most cases with spinal metastases, the quality of life may be improved by vertebral replacement with Harms' titanium cage. Amelioration of clinical symptoms such as neurological deficit, pain, and tack of walking ability occurred in most patients. PMID- 10820886 TI - [Arterial supply of the femoral condyles]. AB - INTRODUCTION: The most frequent site for aseptic bone necrosis and osteochondrosis dissecans in the human knee joint is the medial femoral condyle. The aim of this study is to analyze the three-dimensional vascularization of the human knee joint and to find out if there are any differences in the blood supply of the medial and lateral femoral condyles which may explain the preponderance of aseptic bone necrosis in the medial side. MATERIAL AND METHODS: The femoral arteries of 14 unfixed lower extremities have been injected with epoxy resin. After documentation of the extraosseous course of the blood vessels the bone was macerated with 10% formic acid. The insertions of tendons and ligaments were investigated by light microscopy and immunohistochemistry. RESULTS: The arterial supply of the lateral femoral condyle originates from the upper lateral geniculate artery, the terminal ends of which penetrate the bone from lateral. Branches of the middle geniculate artery reach the bone from the medial wall of the intertubercular notch. The subchondral bone of the medial femoral condyle draws its blood supply mainly from the descending geniculate artery. Branches of the middle geniculate artery penetrate the medial femoral condyle only in the posterior part of the intercondylar fossa. These posterior vessels do not contribute to the supply of the subchondral bone. In the anterior part of the medial wall of the intercondylar fossa there is the wide femoral insertion of the posterior cruciate ligament. Injection techniques and immunohistochemical investigations show that the fibrocartilage of the chondral apophyseal insertion of the posterior cruciate ligament is avascular and acts as banier which impedes blood vessels to penetrate the bony surface in this region. The subchondral bone adjacent to the femoral insertion of the posterior cruciate ligament receives its blood supply from vessels that penetrate the bone at the medial wall of the medial femoral condyle. The blood vessels which supply the lateral parts of the medial condyle have the longest intraosseous course. The density of vessels within this region is largely reduced. CONCLUSIONS: The region adjacent to the femoral insertion of the posterior cruciate ligament is the most frequent site for osteochondrosis dissecans in the knee joint. Our results show that the arterial supply of the subchondral bone may be considered as a cofactor for the etiology of osteochondrosis dissecans. PMID- 10820887 TI - [The course of inflammatory mediators after elective orthopedic interventions]. AB - PURPOSE OF THE STUDY: The aim of the study was to characterize the changes of C reactive protein, erythrocyte sedimentation rate, white blood count and body temperature by serial measurements after different types of uncomplicated orthopedic surgery. METHODS: The uncomplicated course of 180 patients after elective total hip and knee arthroplasties, ventral and dorsal spinal fusions and diagnostic knee and shoulder arthroscopies were analyzed. RESULTS: The maximal CRP-values were recorded on the second or third day after operation. The peak levels correlated with the extent of the procedures and reached 1.28-17.71 mg/dl. The ESR increased to maximal rates five to six days after surgery and remained elevated during the 14 days of the investigation period. WBC and body temperature showed a slow and uncharacteristic increase followed by a slow and irregular decrease. CONCLUSION: Awareness of the typical pattern of CRP, ESR, WBC and body temperature may help to evaluate the early postoperative course. The CRP is a sensitive marker. ESR, WBC and body temperature are less costly but a poor diagnostic aid for the early detection of postoperative complications especially infections. PMID- 10820888 TI - A comparison of vancomycin and sulfated beta-cyclodextrin as chiral selectors for enantiomeric separations of selenoamino acids using capillary electrophoresis with UV absorbance detection. AB - The enantiomeric separation of three selenoamino acids, D,L-selenomethionine, D,L selenoethionine and D,L-selenocystine is described. Both sulfated beta cyclodextrin and vancomycin have been successfully used to separate all enantiomers of the compounds with UV detection. Reproducible separations, in terms of peak area and migration time were obtained using sulfated beta cyclodextrin with reversed polarity and UV detection. With vancomycin as a chiral selector, reversed polarity was found to be more reproducible than positive polarity in terms of peak migration times. PMID- 10820889 TI - High-performance liquid chromatography and inductively coupled plasma mass spectrometry (HPLC-ICP-MS) for the analysis of xenobiotic metabolites in rat urine: application to the metabolites of 4-bromoaniline. AB - The use of HPLC-ICP-MS for the profiling and quantification of the metabolites of 4-bromoaniline following reversed-phase gradient chromatography is demonstrated. In the 0-8 h post dose sample, which contained the highest concentrations of compound-related material, it was possible to detect at least 16 metabolites of the compound. The methodology described offers the possibility of obtaining metabolite profiles and quantification for drugs and other xenobiotics in biological fluids and excreta without the requirement for radiolabelled tracers. PMID- 10820890 TI - Probing protein-peptide-protein molecular architecture by atomic force microscopy and surface plasmon resonance. AB - We demonstrate the creation of a protein multilayer which utilises the high affinity interaction between streptavidin and biotin and incorporates a peptidic spacer. Surface plasmon resonance measurements enabled us to monitor the construction of the multilayer in real time. Atomic force microscopy was utilised to determine surface functionality at each stage of the multilayer construction, allowing us to investigate the associated mechanical properties. In this context we observed an increase in biomolecular stretching on the formation of the multilayer. We demonstrate, utilising circular dichroism, that variations in the solvent can affect the secondary structure of the peptide linker and hence its mechanical properties. Trifluoroethanol titrations on the assembled system indicate that the multilayer properties are also stimuli responsive with regard to solvent conditions. These results indicate that the multilayer stretch before cleavage is increased in the presence of trifluoroethanol. This was not expected from the study of the individual linker alone, indicating the need to study the system as a whole as opposed to the isolated components. PMID- 10820891 TI - Automatic semi-microcolumn liquid chromatographic determination of catecholamines in rat plasma utilizing peroxyoxalate chemiluminescence reaction. AB - A fully automated and highly sensitive method with a semi-microcolumn liquid chromatography system for the determination of rat plasma catecholamines (CAs) was developed. Automated on-line extraction of CAs in diluted plasma using a precolumn packed with strong acidic cation exchange resin was coupled with separation of CAs on a semi-microcolumn (250 x 1.5 mm id). fluorogenic derivatization with ethylenediamine and finally postcolumn peroxyoxalate chemiluminescence detection utilizing bis[2-(3,6,9-trioxadecanyloxycarbonyl)-4 nitrophenyl]oxalate (TDPO) and hydrogen peroxide. The detection limits were 0.91, 0.36 and 1.1 fmol for norepinephrine (noradrenaline), epinephrine (adrenaline) and dopamine, respectively, at a signal-to-noise ratio of 3. A good linearity of the calibration curve for each CA was observed in the range of 5.0 to 500 fmol for each CA using N-methyldopamine (N-MeDA) as an internal standard. The RSD for the proposed method (n = 5) were 3.7-9.5% for the intra-day assay and 6.6-10.0% for the inter-day assay. The volume of rat plasma required for the determination of CAs was 10 microliters. PMID- 10820892 TI - Determination of arsenic in biological fluids by electrothermal atomic absorption spectrometry. AB - A procedure for the determination of the total content of arsenic in urine, serum and blood by electrothermal atomic absorption spectrometry (ETAAS) is described. Zeeman correction is used to compensate the high background signals. The samples are diluted (1 + 1 for urine and 1 + 3 for both serum and blood samples) in a medium containing 0.1% w/v Triton X-100 before being introduced directly into the furnace. A solution containing 15% w/v hydrogen peroxide, 0.65% w/v nitric acid and 0.5% w/v nickel is also introduced into the atomizer by means of a separate injection. Calibration is carried out against aqueous standards for blood and serum samples and using the standard additions method for urine samples. The detection limit is 20 pg (2 ng ml-1). The reliability of the procedure is checked by analyzing three certified reference materials and by recovery studies. PMID- 10820893 TI - Oral contraceptives and cardiovascular risk. AB - In 1992, we advised that "the oral contraceptive of choice for a healthy young woman probably remains a combined preparation containing 30 or 35 micrograms oestrogen and one of the newer progestogens" (i.e. desogestrel, gestodene, norgestimate). In 1995, evidence emerged suggesting that combined oral contraceptives containing desogestrel or gestodene were associated with a risk of venous thromboembolism about twice that of pills containing other progestogens, and the Committee on Safety of Medicines (CSM) advised that desogestrel- and gestodene-containing pills should be used only by women who were intolerant of other combined oral contraceptives and prepared to accept an increased risk of venous thromboembolism. Following an appeal by the drug manufacturers to the UK Medicines Commission, this advice has been modified to state that, provided women are fully informed of the increased risks, "it should be a matter of clinical judgement and personal choice which type of oral contraceptive should be prescribed". Here, we discuss the evidence and events leading to the latest advice, reconsider our previous recommendations, and discuss other cardiovascular problems that may be associated with oral contraceptive pills. PMID- 10820894 TI - The use of inhaled corticosteroids in adults with asthma. AB - Inhaled corticosteroids have now been used for the treatment of asthma for over 25 years. They have largely replaced oral corticosteroids for the prevention of asthma symptoms and asthma attacks, and for most patients they have become the first line of preventive drug therapy. We recently discussed the role of inhaled corticosteroids in the management of childhood asthma. Here, we review their place in the management of adults with asthma. A forthcoming article will review the various types of inhaler now available for delivering drugs for asthma. PMID- 10820895 TI - Thioacetamide toxicity and the spleen: histological and biochemical analysis. AB - Exposure to thioacetamide is associated with the development of liver cirrhosis in experimental animals. In addition to liver, thioacetamide toxicity has been observed in other organs. In this study, the toxic effect of thioacetamide on the spleen was investigated at 0, 4, 8 and 12 weeks post-treatment durations. The level of tissue copper and selenium increased until the eighth week when a significant drop was observed. The zinc level was also increased but returned back to normal by week 8, thereafter it showed further increase. Calculation of the copper/zinc ratio showed an increase, but, recovered and returned to normal value by week 12. The level of manganese fluctuated until the eighth week. It then increased rapidly. Histological studies of the spleen tissue showed a significant increase in extramedullary haematopoiesis in the red pulp region and marked hyperplasia in the marginal zone and follicles. The results of this study, demonstrate an intimate association between trace element levels and spleen pathology, as observed in studies of other organs. PMID- 10820896 TI - Immunohistochemistry of carbonic anhydrase isozymes (CA-I, II and III) in canine salivary glands: a distributional and comparative assessment. AB - The immunohistolocalization of carbonic anhydrase isozymes (CA-I, II, III) in canine salivary glands was studied using antiserum against CA-I, II, III. In parotid glands, immunostaining intensely localized cytosolic CA-II antiserum throughout the cytoplasm of acinar secretory cells and ductal epithelial cells, especially in the striated duct region. CA-III reactivity in the glands was only seen selectively at the intercalated ductal cells. In contrast, no immunoreaction localized CA-I in the gland. In the submandibular and sublingual glands, CA-I, II, and III were all observed in the ductal segments of the glands, whereas serous demilune appeared devoid of all three cytosolic CA isozymes. In contrast, in zygomatic glands (i.e. dorsal buccal glands) all CA isozymes were observed in both serous demilune and ductal segments. In all of the salivary glands examined, no mucous acinar cells were found to be reactive for any CA. PMID- 10820897 TI - Light and ultrastructural immunocytochemical study of somatotropic cells (GH cells) in ovine adenohypophysis: lactation and weaning influences. AB - The influence of lactation period and weaning on the distribution, number, and structure of somatotropic cells (GH cells) in ewes was studied using immunocytochemical procedures for light and electron microscopy, as well as morphometric and stereological techniques. The adenohypophyseal gland of 12 ewes of the Segurena breed in different stages of milk production and weaning was studied, while three ewes in anoestrus served as controls. The size of secretory granules was heterogeneous in all stages, suggesting that this characteristic is not related to functional activity. During lactation and weaning the size of GH cells decreased, while the number of 'synthesis cells' increased. The rough endoplasmic reticulum and Golgi complex appeared more developed and small secretory granules showed lower gold particle labelling. These data suggest that GH cells are more active during lactation, particularly during late lactation. PMID- 10820898 TI - Complex carbohydrate histochemistry and ultracytochemistry of the sheep lacrimal gland. AB - The chemical content of the secretion of the sheep lacrimal gland was analysed at the light and electron microscope levels by applying histochemical techniques and an ultrastructural histochemical method (periodic acid, thiocarbohydrazide and silver proteinate). Mucosubstance histochemistry demonstrated acidic glycoconjugates, mainly sulphated, in the mucous and seromucous glandular cells and in the apical portion of the cells lining the terminal ducts. Moreover, secretory granules, stained with PA-TCH-SP, showed a different localization of the reaction product. The presence of lysozyme was also found in the glandular serous cells. These histochemical studies demonstrate that the secretion of sheep lacrimal glands is mixed, having serous, mucous and seromucous components, and that an excellent correlation exists between the secretory granule substructure and glycoprotein localization. PMID- 10820899 TI - Anatomical evaluation of the caprine mammary gland by computed tomography, radiology and histology. AB - An anatomical study of the mammary gland by computed tomography, X-rays and histology was carried out. Three canarian breed goats were used. Identification of the udder structures by different methods represented the basis of this study. Using these techniques the internal and external organization of the caprine mammary gland was studied. PMID- 10820900 TI - An anatomical study of the arteries feeding the triceps brachii muscle of swine. AB - The arterial vascular network of the porcine triceps brachii muscle (TBM) (an extensor muscle to the forearm) was studied and compared to another extensor muscle (the soleus muscle) of small rodents. The left axillary arteries (LAA) of nine Yucatan miniature swine were perfused with latex material to reveal the organization of the arterial blood supply to the TBM. Blood is supplied to the TBM by the main branches of the LAA. Some of the branches end in one of the four heads of the TBM. Other branches continue to bifurcate further and to supply blood to adjacent muscles. The feed arteries (FAs) arise as side-branches at regular distances from the branches of the LAA. The mean number of FAs per TBM was 109.7 and the mean diameter was 388 microns. The distribution of FAs to the muscle area is heterogenous. Most FAs penetrate the epimysium of the TBM on its medial aspect. It appears that more FAs come out of the branches of LAA that end in the TBM as compared to arteries which continue to bifurcate away from TBM. FA diameters were in the range of 300 microns. This study is the first necessary step towards understanding the distribution of oxygen and nutrients to the porcine skeletal muscle. It reveals the presence of a more complicated vascular network than that observed previously in small laboratory animals. PMID- 10820901 TI - Ultrastructure of the horse tongue: further observations on the lingual integumentary architecture. AB - This investigation examined primarily epidermal specializations of the adult horse tongue by light, scanning and transmission electron microscopy. Samples were collected from seven regions of the normal tongue of various breeds of horse. The filiform papillae, present on the dorsal and lateral aspects but not the ventral aspect of the tongue, were short, slender and finger-like structures with variable-shaped terminae. The epidermal thickness and height of dermal ridges were reduced on fungiform and vallate papillae, but tissue architecture and keratinocyte ultrastructure of most of the lingual epidermis corresponded to the common mammalian epidermal paradigm. One unique finding was the highly localized clustering of epidermal cells with exceptionally high content of PAS negative trichohyalin cytoplasmic granules at a location atop the dermal ridges and beneath the base of filiform papillae. These granular cells were immediately subjacent to clusters of clear, non-granulated epidermal cells. It is believed that this integumentary specialization may enhance the structural strength at this localized site of the tissue architecture, in relationship to the mechanical papillae. PMID- 10820902 TI - Various types of glomus anastomoses in developing rat fetuses. AB - Some observations on arteriovenous anastomosis (AVA) morphogenesis of rats were considered in this investigation. Interesting typical glomus anastomoses observed under light microscopy were determined by photography before evaluation. AVAs in the skin and mesentery were compared with those reported in morphological studies in the literature. Most of the glomus anastomoses were determined to be in conformity with those in the studies reported. Some resembled simple shunts and others complex glomus anastomoses. It is suggested that the existence of various types of anastomoses of rat fetuses during development is dependent on blood flow. PMID- 10820903 TI - Suspensory apparatus of the bovine udder in the Estonian black and white Holstein breed: increased milk production (udder mass) induced changes in the pelvic structure. AB - Bovine udders, together with the attached section of the ventral abdominal wall, as well as the bony pelvic floor were studied. The parts of the suspensory apparatus and attachment to the body were investigated. The location of the medial and lateral laminae served as the basis for classifying them into abdominal and pelvic parts. The udder base is attached to the ventral abdominal wall by means of the dorsal connective tissue plate. The abdominal part of the medial lamina and the pelvic part of the lateral lamina are the functionally more important bands. Sixty per cent of the udder base is attached to the ventral abdominal wall and 40% to the pelvic floor by the symphysial tendon. The symphysial tendon is attached to the ventral pubic tubercle and the symphysial eminence as well as to the symphysial crest, which develops by the fourth year. In order to study the osseous changes, bovine pelvises from the Middle Ages, the beginning of the 20th century, and the present time were compared. Increased udder mass has brought about changes in the structures of the suspensory apparatus of the udder and in the structure of the pelvis. PMID- 10820904 TI - Establishment of an animal model of hepatic encephalopathy. AB - BACKGROUND: Hepatic encephalopathy is a neuropsychiatric syndrome associated with acute liver failure, chronic parenchymal liver disease or portal systemic anastomosis. The spectrum of symptoms may vary from subtle mental changes to a disrupted circadian rhythm to hepatic coma. In order to investigate the possible pathogenetic mediators and the use of potential new therapies, the aim of our study was to create a reliable animal model for research on hepatic encephalopathy. METHODS: Forty male Sprague-Dawley rats were used. Fulminant hepatic failure was induced by intraperitoneal injection of thioacetamide (TAA, 350 mg/kg) for three consecutive days (n = 30). Rats treated with normal saline served as controls (n = 10). The clinical stage of hepatic encephalopathy in TAA treated rats was graded according to neurobehavioral testing. Spontaneous motor activities of rats were calculated using the Opto-Varimex animal activity meter. We also investigated the relationships between the neurobehavioral score and the motor activity count. RESULTS: Compared with normal, saline-treated rats, those receiving consecutive injections of TAA had apparently lower neurobehavioral scores (p < 0.001) accompanied by significantly blunted motor activities (p < 0.001). A significant correlation was demonstrated between the neurobehavioral score and total movements in rats with encephalopathy (r = 0.71, p < 0.001). In addition, the neurobehavioral score also correlated well with ambulatory movements (r = 0.73, p < 0.001) and vertical movements (r = 0.45, p < 0.05). CONCLUSIONS: The rat model is acceptable for the study of hepatic encephalopathy, as established in this study. This experimental model can be used to explore the pathogenesis and management of hepatic encephalopathy. PMID- 10820905 TI - Analysis of lipids in non-diabetic patients with acute myocardial infarction. AB - BACKGROUND: The incidence of postinfarct cardiac events can be reduced through secondary prevention by lipid regulation. The relationship between early-detected lipids and prognosis was investigated prospectively in 97 non-diabetic patients with acute myocardial infarction (AMI). METHODS: Blood samples were analyzed in five evolving stages of AMI: 1) immediately after admission (< 24 hours after the onset of symptoms); 2) on the second day after admission (< 48 hours after the onset of symptoms); 3) on the seventh day after admission; 4) two weeks after the AMI; and 5) three months after the AMI. Cardiac events, including congestive heart failure, reinfarction, unstable angina, ventricular tachyarrhythmia and sudden cardiac death were evaluated at a mean follow-up of 26 months. RESULTS: Serial measurements in 75 cases with complete follow-up showed that all lipids except lipoprotein(a) had a decline in plasma level after patients were admitted to hospital. The concentrations of lipids three months postinfarct approached the admission values. Age, body mass index, vessel number, severity of vessel disease and the initial values of lipids on admission had no influence on postinfarct cardiac events in these patients. CONCLUSIONS: The results indicate that the plasma levels of lipids detected within 24 hours after AMI can be used as the baseline lipid levels. Nevertheless, the impact of these lipids on the adverse outcomes in non-diabetic AMI patients should be further studied in a large-scale study. PMID- 10820906 TI - Impact of Helicobacter pylori eradication on the development of MALT, gland atrophy and intestinal metaplasia of the antrum. AB - BACKGROUND: Chronic Helicobacter pylori infection leads to the development of mucosa-associated lymphoid tissue (MALT), gland atrophy, intestinal metaplasia, and in certain individuals, gastric lymphoma and adenocarcinoma. We conducted a prospective study to determine the effect of H pylori eradication on the development of MALT, gland atrophy and intestinal metaplasia. METHODS: From July 1995 to October 1996, 63 consecutive patients with H pylori-associated peptic ulcer disease were enrolled in a bacterial eradication trial. The grade of MALT and the degree of gastritis of the antrum were assessed using Wotherspoon's scale and the Sydney system before treatment, and at eight weeks and one year after H pylori eradication therapy. RESULTS: During the study period, 43 patients received complete follow-up. In the patient group with eradication failure (n = 22), MALT score was significantly decreased at the end of the eighth week (p < 0.05) but returned to the initial level by one year of follow-up. There were no changes in the scores for inflammation, neutrophil activity, gland atrophy or intestinal metaplasia at the end of the eighth week and at one year following H pylori eradication therapy. In contrast, there was a marked reduction in the MALT, inflammation, and activity scores at eight weeks (p < 0.01, < 0.05 and < 0.05, respectively) and one year after treatment (p < 0.05, < 0.001 and < 0.001, respectively) in the patient group with successful eradication (n = 21). However, no significant changes in gland atrophy and intestinal metaplasia were observed during the follow-up period. CONCLUSIONS: Eradication of H pylori leads to regression of MALT in the stomach, but the degrees of gland atrophy and intestinal metaplasia remain unchanged, even after one-year of follow-up. PMID- 10820907 TI - Prevalence of the Taiwan variant of the Epstein-Barr virus in nasopharyngeal carcinoma patients and normal individuals. AB - BACKGROUND: Latent membrane protein 1 (LMP1) of Epstein-Barr virus (EBV) is a potentially oncogenic protein, able to inhibit differentiation and induce transformation of epithelial cells. The LMP1 gene sequence of EBV in nasopharyngeal carcinoma (NPC) differs from that of EBV in B95-8 cells. A change of nucleotide at the XhoI site results in loss of the restriction site in the EBV LMP1 gene from NPC, and this of EBV is designated as the Taiwan variant. In this study, we further investigated the Taiwan variant of EBV in nasopharyngeal biopsies and throat washings of NPC patients and normal controls. METHODS: DNA was extracted from nasopharyngeal biopsies of 22 NPC patients and 40 normal controls, and from throat washings of 28 NPC patients and 78 normal controls. The DNA was amplified by polymerase chain reaction (PCR) using LMP1 gene-specific primers, then the EBV variant was identified by analysis with the XhoI restriction enzyme. RESULTS: The LMP1 gene was detected in all NPC tumors, in only 25 of 40 normal nasopharyngeal biopsies, and in 12 of 28 and 44 of 78, respectively, throat washings of NPC patients and normal controls. The Taiwan variant was detected in 84.1% to 100% of EBV isolates. The distribution of Taiwan variant EBV was not statistically different between the NPC group and normal controls in either nasopharyngeal biopsies or throat washings. CONCLUSIONS: The results indicate that, based on XhoI restriction site analysis, the Taiwan variant of EBV is the predominant EBV strain in NPC patients and normal individuals in Taiwan. PMID- 10820909 TI - Endoscopic endonasal transsphenoidal surgery for pituitary tumors. AB - BACKGROUND: The traditional transsphenoidal approach for pituitary surgery can be performed via transnasal, transseptal, or sublabial routes through unilateral or bilateral nostrils. It requires wide mucosal and septal dissection and postoperative nasal packing. Endoscopic surgery has been widely used because it allows excellent visualization with minimal invasion. Recently, it has also been applied to pituitary surgery. METHODS: From January, 1997 to February, 1999, 40 patients with pituitary adenomas underwent the transsphenoidal approach using a rigid endoscope via one nostril. Among the 40 cases, 18 were prolactinomas, seven were growth hormone-secreting adenomas and 15 were nonfunctional adenomas. RESULTS: After surgery, complete resolution or improvement of symptoms and restoration of normal hormone levels were achieved in 16 patients with prolactinomas, five with growth hormone-secreting adenomas and 12 with nonfunctional macroadenomas. One patient with a recurrent microprolactinoma needed a second operation to remove the cavernous portion of the tumor. Another male patient with a macroprolactinoma who experienced galactorrhea and gynecomastia showed improvement of clinical symptoms after the operation. Two patients with residual growth hormone-secreting macroadenomas in the cavernous sinus needed Sandostatin treatment. Three patients with nonfunctional macroadenomas underwent nearly total resection leaving residual tumor in the cavernous sinus, which then required adjuvant radiotherapy. No complications related to this approach were encountered in the patients during the follow-up period. CONCLUSIONS: Endonasal transsphenoidal endoscopic surgery can be employed for treating pituitary tumors without septal or sublabial complications. Postoperative suffering was reduced and hospitalization was shortened by this mini-invasive procedure. This surgical procedure can be used for both microadenomas and macroadenomas. PMID- 10820908 TI - Effect of milrinone on postbypass pulmonary hypertension in children after tetralogy of Fallot repair. AB - BACKGROUND: Postbypass pulmonary hypertension in surgical correction of tetralogy of Fallot (TOF) is a risk for right ventricular failure. Effective management remains a major challenge. Milrinone is a new drug with a unique mechanism of "inodilation", which offers both inotropic and vasodilatory effects. We attempted to determine if application of milrinone could improve cardiopulmonary dysfunction in children after TOF repair. METHODS: We studied 10 children with postbypass pulmonary hypertension after TOF repair within six months. Heart rate, systolic pulmonary arterial pressure (PAP), systolic arterial blood pressure (SBP), pulmonary capillary wedge pressure and PAP/SBP ratio were recorded. Standard cardiopulmonary bypass (CPB) was performed. After CPB, if PAP/SBP was more than 0.5, pulmonary hypertension was suspected and milrinone was administered with a loading dose of 20 micrograms/kg followed by continuous infusion of 0.2 microgram/kg/minute. Hemodynamics were compared before and after administration of milrinone to evaluate its effect. RESULTS: significant reduction in PAP/SBP ratio within 15 minutes was found after administration of milrinone. The effect persisted for 24 hours during continuous infusion of milrinone. No remarkable adverse effect was noted in the study. CONCLUSIONS: We conclude that milrinone is effective in the management of pulmonary hypertension following CPB in children who underwent TOF repair. PMID- 10820910 TI - Changes in cytokine (IL-8, IL-6 and TNF-alpha) levels in the amniotic fluid and maternal serum in patients with premature rupture of the membranes. AB - PURPOSE: To study the changes in interleukin-6 (IL-6), interleukin-8 (IL-8) and tumor necrosis factor-alpha (TNF-alpha) levels in patients with premature rupture of the membranes (PROM). METHODS: A total of 46 normal term pregnant women served as controls and 46 women with PROM were enrolled for study. Maternal serum and amniotic fluid IL-6, IL-8 and TNF-alpha levels were determined by enzyme-linked immunosorbent assay and a sensitive radioimmunoassay, respectively. RESULTS: The maternal serum IL-6 and IL-8 levels and amniotic fluid IL-6, IL-8 and TNF-alpha levels were higher than those of controls, and the differences were significant between the two groups (p < 0.05). Although the levels of TNF-alpha in maternal serum were higher than that of controls, the difference was not significant. There were significant relationships between the levels of IL-6, IL-8 and TNF alpha and the duration of PROM. The longer the duration of PROM, the higher the levels of maternal serum and amniotic fluid IL-6, IL-8 and TNF-alpha were. There were 12 cases of clinical chorioamnionitis, and the levels of IL-6, IL-8 and TNF alpha in the maternal and amniotic fluid were higher than that of those without chorioamnionitis (p < 0.05). CONCLUSION: Determining cytokines (IL-6, IL-8 and TNF-alpha) is a valuable clinical method for identifying chorioamnionitis in patients with PROM. PMID- 10820911 TI - Mah-jong epilepsy: a new reflex epilepsy. AB - BACKGROUND: Flickering light and color patterns, reading, language, movement, decision making, eating, tapping and touching, hot water immersion and auditory stimulation can induce seizures in some patients. This is known as "reflex epilepsy". The mechanism of reflex epilepsy is not clear. Recently, we cared for 12 patients (11 men and 1 woman, age 41-74 years; mean age = 63.1 years) whose seizures were induced by playing mah-jong, with mean age at seizure onset of 48.7 years. Nine of the 12 patients had seizures exclusively while playing mah-jong. METHODS: We retrospectively reviewed the medical records of 12 epileptic patients who visited our neurologic clinic from 1987 to 1999, with the chief complaint of mah-jong induced seizures. All patients underwent examinations including electroencephalography, brain computed tomography or magnetic resonance imaging, analysis of clinical manifestation of seizures, family history and past medical history. All were given anticonvulsant therapy for preventing seizures. RESULTS: Clinically, 10 patients had generalized tonic-clonic seizures and two patients had partial seizures with secondary generalization. Interictally, normal results on EEG were found in six patients, three had focal temporal spikes, and three had intermittent slow activity in the frontotemporal regions. Neuroimaging studies of the brain were normal in seven patients, two had lacunar infarctions, one had generalized atrophy and one had focal left parietal lesions of an unclear nature. Only one patient had a family history of epilepsy. All 12 patients received anticonvulsant therapy, and of these, nine had good epilepsy control. CONCLUSIONS: "Mah-jong epilepsy," a new reflex epilepsy, is probably related to thinking and decision making while playing mah-jong. The easy control of seizures induced by mah-jong suggests a benign nature. In addition to anticonvulsant therapy, avoiding playing mah-jong may be essential in preventing seizures. PMID- 10820912 TI - Minimally invasive mitral valve surgery via submammary hidden incision. AB - BACKGROUND: With the recent advancement in minimally invasive cardiac surgery based on the needs of decreasing costs, hospital stay, recovery time and improving cosmetic appearance, minimally invasive mitral valve surgery (MIMS) is being actively pursued. The objective of this study was to compare our submammary hidden incision method to two other more proven methods of MIMS with respect to length of operation, clinical outcome and cosmetic satisfaction. METHODS: From July, 1996 to June, 1998, a total of 14 patients underwent MIMS in our hospital. Three different approaches were used: the limited right-side thoracotomy approach, the parasternal incision approach and our submammary hidden incision approach, of which, this is the first report. All relevant clinical results are reported, including general characteristics of the operations, morbidity and mortality, length of hospitalization, pain relief and patient satisfaction. RESULTS: Five patients underwent MIMS via limited thoracotomy, four via parasternotomy and five with our submammary hidden incision method. There were six mitral valves repaired and eight replaced. Conversion to traditional sternotomy was not necessary in any of the three groups. The results from the three different approaches did not differ significantly across the different parameters measured. Nonetheless, the submammary hidden incision approach showed promise in the areas of length of stay for both intensive care patients (36.3 +/- 5.0 hours) and total hospitalization (10.7 +/- 0.6 days), pain relief and patient satisfaction. CONCLUSIONS: The submammary hidden incision method does not result in any significant disadvantage in clinical outcome. The method resulted in more satisfied patients due to less pain, shorter hospital stay, and most importantly, cosmetic appearance of the resulting surgical scar. We will make the submammary hidden incision approach the MIMS approach of choice at our institution and continue to monitor the long-term results. PMID- 10820913 TI - Unusual signs for dural arteriovenous fistulas with diffuse basal ganglia and cerebral calcification. AB - We present a case of multiple dural arteriovenous fistulas (AVFs) in a 60-year old man with the chief complaint of worsening headache, altered mental status and progressively unsteady gait over the course of one year. Computerized tomography revealed diffuse, symmetric calcification in the bilateral basal ganglia and bilateral periventricular and subcortical white matter. Magnetic resonance imaging revealed multiple, enhanced, punctate and linear vessels. These images were due to reflux into the parenchymal veins in the dural AVF of the superior sagittal sinus within the basal ganglia and deep white matter of both cerebral hemispheres. Cerebral angiography disclosed multiple dural AVFs. The exact mechanism of basal ganglia and subcortical calcification is proposed to be an arterial steal phenomenon or persistent venous congestion, with calcification occurring in a chronic hypoperfused state or with dystrophic changes in the walls of congested veins. PMID- 10820914 TI - Ectopic lacrimal gland cyst of the orbit. AB - Lacrimal duct cysts are not common. It is extremely rare when a lacrimal duct cyst and an ectopic lacrimal gland develop in the orbital cavity. A unique case of an ectopic lacrimal gland cyst of the orbit is presented. A 33-year-old man had a palpable mass above the inferior medial orbital rim for nearly two years. An ocular examination was normal except for a movable, firm mass found in the anterior nasal inferior orbit of the right eye. An echogram revealed a homogeneous, hypoechoic cystic mass. Computed tomography of the orbit showed a well-encapsulated lesion in the lower orbit of the right eye near the inferior rectus muscle, without bony erosion. A tense, thin-walled, clear fluid-filled cyst measuring 15 x 12 x 13 mm in size was completely enucleated without rupture by anterior orbitotomy. Pathologic examination disclosed a small nest of normal gland tissue surrounded by a cystic lesion lined with two layers of lacrimal duct epithelium cells. No recurrent signs were noticed during a 12-month period of follow-up. PMID- 10820915 TI - Recurrent gastrointestinal bleeding and high output cardiac failure caused by hereditary hemorrhagic telangiectasia. AB - Hereditary hemorrhagic telangiectasia, commonly known as Osler-Weber-Rendu disease, is a systemic autosomal dominant inherited disorder, that occurs in Caucasian populations. We report the case of a 56-year-old housewife who was admitted to the gastrointestinal and cardiovascular ward because she had suffered from recurrent gastrointestinal bleeding and heart failure from 1994 to 1997. Panendoscopy showed vascular ectasia scattered over the tongue, larynx, esophagus, and posterior wall of the gastric body. Colonoscopy showed clusters of telangiectasia over the cecum and ascending colon. Arteriovenous malformations (AVMs) were found in the liver and lungs on computerized tomography. Recurrent gastrointestinal bleeding was controlled by estrogen treatment during the follow up period. In July 1997, the patient was readmitted to our cardiovascular section due to aggravated dyspnea, orthopnea and bilateral lower leg edema. Cardiac catheterization showed a large fistula from the left pulmonary artery to the left atrium and left ventricle, pulmonary arterial pressure of 37/13 mmHg and cardiac output of 9.61/minute. Other studies excluded the possibility of sepsis, and high output cardiac failure was suspected. The patient was discharged in a stable condition and scheduled for AVM embolization management. Unfortunately, she died of a suspected heart attack at home two weeks following discharge. PMID- 10820916 TI - Malignant mixed mullerian tumors of the ovary. AB - Malignant mixed Mullerian tumor (MMMT) of the ovary is very rare, and to the best of our knowledge, only a few cases have been reported in the literature from Taiwan. We report two recent cases of ovarian MMMT at our hospital. Case 1 was a 59-year-old female with stage IIIC MMMT of the ovary, with a tumor having carcinomatous and sarcomatous elements. The carcinomatous component was composed of a high-grade epithelial malignancy including serous, endometrioid, clear cell and undifferentiated carcinoma elements. The sarcomatous component was composed of a homologous malignant mesenchymal element with conspicuous hyaline globules. The patient died of the disease six months after debulking surgery. Case 2 was a 42-year-old female with ovarian stage IIC MMMT. The carcinomatous component was composed of grade II-III clear cell carcinoma and the sarcomatous component was composed of high-grade non-specific spindle cell sarcoma, which was positive for vimentin, but negative for cytokeratin, desmin and S-100 protein on immunostaining. The patient died of the disease four months after debulking surgery. These two patients both underwent hysterectomy, bilateral salpingo oophorectomy and omentectomy and both received platinum-based chemotherapy after debulking surgery. PMID- 10820917 TI - Rethinking functional limitation pathways. AB - Functional limitation has received considerable attention in gerontology and geriatrics. Much of this work has focused on single-wave transitions devoid of context rather than on the pattern of transitions over time that constitute trajectories. This Forum article suggests that it is time for a different way of looking at functional limitation pathways. It focuses on trajectories. Responses to three Rosow and Breslau (1966) and two Nagi (1976) items, asked of 12,998 older adults who participated in up to seven waves of data collection as part of the Established Populations for the Epidemiologic Study of the Elderly, are used to illustrate this approach, emphasizing both its conceptual and pragmatic advantages. The results provide greater clarity in terms of those who become functionally limited, take on more functional limitations, or recover as well as those who are likely to be lost to follow-up and in terms of the outcomes associated with those individuals over time. PMID- 10820918 TI - Gender differences in psychiatric morbidity among family caregivers: a review and analysis. AB - The major goal of this article was to review and synthesize the empirical research on caregiver gender and psychiatric morbidity, with the aim of answering three questions: (a) Is there greater psychiatric morbidity among female than male caregivers, (b) is the excess psychiatric morbidity among female caregivers attributable to caregiving, and (c) what factors in the caregiving situation contribute to the excess psychiatric morbidity among female caregivers? In almost all studies reviewed, women caregivers reported more psychiatric symptoms than men caregivers. Comparisons with noncaregiving community samples suggest that female caregivers experience excess psychiatric morbidity attributable to caregiving. Using a stress process model as an organizing framework, the study demonstrated that at all stages of the stress process, women are at greater risk for psychiatric morbidity than men. Directions for future research and implications for interventions and public policy are discussed. PMID- 10820919 TI - The dynamics of caregiving: transitions during a three-year prospective study. AB - This prospective study (n = 476) examined 3 types of caregiving transitions experienced by wives and daughters of older persons: entry, institutionalization, and bereavement. Daughters were more likely to enter the caregiving role than wives, but the impact of entering the role was more pronounced for wives. After becoming a caregiver, wives decreased in their participation in leisure activities, perceptions of quality of family relations, and marital satisfaction. Daughter caregivers were more likely than wives to place their care recipient in an institution, and they increased in social participation and decreased in subjective burden after placement. Roughly the same percentage of wife and daughter caregivers were bereaved during the study period, and for wives bereavement was accompanied by an increase in social involvement and personal growth. The results underscore the highly dynamic nature of the caregiving career and the importance of the kinship relationship between caregiver and care recipient in conditioning the effects of caregiving transitions. PMID- 10820920 TI - Matrilineal advantage in grandchild-grandparent relations. AB - This study examines the sources of matrilineal advantage in grandchild grandparent relations using data from the Iowa Youth and Families Project. Results from fixed-effect models indicate that the observed matrilineal advantage in grandchild-grandparent ties arises from lineage differentials in the quality of relations between grandparents and the parents of grandchildren. Specifically, better relations between mothers and the maternal line facilitate closer ties between grandchildren and maternal grandparents. Fathers' closer ties with the paternal side also promote better relations between a grandchild and paternal grandparents, but the greater prevalence of matrilineal bias in parent grandparent ties leads to an overall matrilineal advantage in grandchild grandparent relations. PMID- 10820921 TI - Goals in geriatric assessment: are we measuring the right outcomes? AB - Previous evaluative studies of outpatient geriatric assessment have focused on a limited set of outcomes related to functioning, health services utilization, and costs. The purpose of this study was to identify important goals for care as described by patients and family caregivers being cared for in this setting. Using a cross-section of 226 consecutive sets of patients and their primary family caregivers, physicians, and case managers, goals of care for individual patients were coded from open-ended interview responses. The most common categories of goals expressed by family caregivers were obtaining education and referrals (57.5%) and improving social and family relationships (53.0%). The process of establishing and meeting such goals should be explicitly included in the design of future evaluations of outpatient geriatric assessment. PMID- 10820922 TI - Managed care and multilevel long-term care providers: reluctant partners. AB - Managed care is reshaping our health care system, although long-term care is only beginning to feel its effects. We report on the managed care involvement of 492 multilevel, long-term care facilities (MLFs; including skilled nursing and assisted/independent living) nationally. Organizational structure and culture and especially environmental characteristics are associated with whether facilities have contracts with managed care organizations (MCOs), plan to have contracts, are only gathering information on MCOs, or intend to do nothing in the near future. Resource dependence theory best explains MCO contracting patterns with MLFs appearing to be responding more to survival than to growth. PMID- 10820923 TI - Reality orientation for dementia: a systematic review of the evidence of effectiveness from randomized controlled trials. AB - The effectiveness of classroom reality orientation (RO) in dementia was evaluated by conducting a systematic literature review. This yielded 43 studies, of which 6 were randomized controlled trials meeting the inclusion criteria (containing 125 subjects.) Results were subjected to meta-analysis. Effects on cognition and behavior were significant in favor of treatment (cognition standardized mean difference [SMD] = -0.59; 95% confidence interval [CI] -0.95(-)-0.22; behavior SMD = -0.64, 95% CI = -1.20(-)-0.08). The evidence indicates that RO has benefits on both cognition and behavior for dementia sufferers. However, a continued program may be needed to sustain potential benefits. Future research should evaluate RO in well-designed multicenter trials. PMID- 10820924 TI - Implementation of advance directives among community-dwelling veterans. AB - To evaluate the feasibility and effectiveness of implementing a "Let Me Decide" advance directive education program among veterans living in the community, the authors studied 150 veterans in south central Ontario. Thirty-four veterans had preexisting Powers of Attorney and were removed from the analysis, leaving a total sample of 116. Two methods of systematically implementing a directive program were evaluated after the intervention period and 6 months later. Eighty two (71%) of the 116 veterans expressed interest in receiving detailed information about the program, and 67 (82%) of the 82 interested veterans were educated. Forty-two (63%) of the 67 educated veterans completed directives. Of the 116 interested veterans, 42 (36%) completed directives. Veterans who were educated about directives were surveyed at follow-up, and 37 of 38 (97%) respondents reported that the education process was beneficial and should be offered to other veterans. This response pattern was consistent among those who completed and those who did not complete directives. PMID- 10820925 TI - Participants in the Program of All-Inclusive Care for the Elderly (PACE) demonstration: developing disease-impairment-disability profiles. AB - The Program of All-Inclusive Care for the Elderly (PACE) builds on On Lok's community-based care and financing model for disabled elderly people who are state certified as eligible for nursing home care. Yet PACE's diverse population has not been fully described. We obtained data for a complete cross-section of PACE participants from early 1997 (N = 2,917). Using grade-of-membership analysis, we classified participants on the basis of their specific diseases, impairments, and disabilities. The classification was reviewed by a physician panel to produce clinical profiles, which were then validated against participants' PACE tenure, demographics, supports, and health. Cognitive impairment, incontinence, and activities of daily living disabilities were influential in producing eight types, which correspond predictably to responses in tenure (the more disabled, ill types likely to be in PACE longer), demographics, health, and informal support. PMID- 10820926 TI - Social contact, socioeconomic status, and the health status of older Malaysians. AB - We tested the applicability of the stress buffering hypothesis in a developing country setting with data from the Senior Sample of the Malaysian Family Life Survey-2. Using ordered logistic regression methods, we examined whether having daily contact with adult children moderates the effect of low socioeconomic status (SES; conceptualized as a chronic stressor) on self-assessed health status. We found that low SES is associated with poorer health for all three ethnic groups--Malay, Chinese, and Indian. Further, for Malays and Chinese, we found that the negative effects of low SES on health tend to be stronger for older people with less frequent contact with adult children than for those who have daily contact. These results provide general support for the buffering model and suggest that, as found in developed countries, active intergenerational relationships in developing country settings may have protective effects on the health of older people experiencing chronic stressors. PMID- 10820927 TI - Health contract calendars: a tool for health professionals with older adults. AB - A modification of the health contract technique was applied by 4 geriatric fellows from the Center on Aging, University of Texas Medical Branch, and 3 family medicine residents from the Department of Family Medicine, University of Texas Medical Branch, to a diverse group of 48 older adults. The innovation, a calendar component to the health contract, allowed for the calculation of specific success rates. Fifteen clients had a 100% success rate, and 21 were highly successful, 8 not too successful, and 4 unsuccessful. PMID- 10820928 TI - Angiotensin II type 1 receptor blocking agents in regression of cardiac hypertrophy--how and why? PMID- 10820929 TI - Influence of total cholesterol levels on long-term mortality in coronary heart disease: a reappraisal. AB - To examine the prognostic significance of total cholesterol levels at baseline in subjects with stable coronary heart disease, 605 patients with stable coronary heart disease were enrolled; 45 of these did not meet inclusion criteria, 41 were lost to follow-up and 40 opted for coronary bypass surgery. Data of the remaining 479 (389 males, 90 females) were analysed. There were 102 males in group I (cholesterol < 200 mg/dL), 187 in group II (cholesterol 200-239 mg/dL), and 100 in group III (cholesterol > or = 240 mg/dL) and 49 females in group I and 41 in group II. The groups were evenly matched for age and numbers with stable angina or survivors of myocardial infarction. Proportion of smokers, hypertensives, diabetics or obese was also similar (p > 0.05). Mean follow-up in years in men was 6.82 +/- 3.15 in group I, 6.37 +/- 3.11 in group II and 6.81 +/- 2.84 in group III while in women it was 6.95 +/- 2.84 in group I and 7.03 +/- 2.58 years in group II and was not different in various groups (p > 0.05). The overall cardiovascular mortality in various groups in men was 20.6 percent in group I, 28.9 percent in group II and 23.0 percent in group III and in women it was 14.3 percent in group I and 22.0 percent in group II. The crude mortality rate was 2.51 percent per year in males and 1.77 percent per year in females. Actuarial survival at end of seven years in males was 0.76 +/- 0.05 in group I, 0.67 +/- 0.04 in group II, and 0.67 +/- 0.05 in group III and in females it was 0.85 +/- 0.05 in group I and 0.73 +/- 0.09 in group II. The cumulative hazard rates per 1000 person- year follow-up in group I, II and III in males were, at age less than 50 years: 5.4 +/- 5, 19.8 +/- 7, 17.4 +/- 8; at 50-59 years: 23.8 +/- 11, 38.5 +/- 9, 39.8 +/- 13; and at 60 years and over: 76.9 +/- 20, 112.6 +/- 20, 108.2 +/- 28, respectively (p < 0.001 on comparison of group I with groups II and III). In females the trends were not significant. Total cholesterol levels at baseline predict long-term cardiovascular mortality in men with stable coronary heart disease. PMID- 10820930 TI - High risk for coronary heart disease in Thiruvananthapuram city: a study of serum lipids and other risk factors. AB - There is a trend towards increase in the incidence of coronary heart disease among Indian population. Also, little information is available on the population distribution of serum lipid components and risk factors for coronary heart disease in Kerala, a state fast turning urban. To study the serum lipid profile and the prevalence of other risk factors for coronary heart disease in the residents of an urban housing settlement in Thiruvananthapuram, fasting blood sample was collected from 206 (64%) residents above the age of 19 years and analysed for plasma glucose and various fractions of serum lipids. A detailed questionnaire on the clinical profile and history of the subjects, and measured weights and heights was also administered. Mean serum total cholesterol was 223.7 +/- 45.3 mg/dL; 223.7 +/- 44.9 mg/dL among males and 223.7 +/- 45.8 mg/dL among females. Mean high-density lipoprotein cholesterol was consistently higher in females in all age groups, while mean low-density lipoprotein cholesterol was higher in males till the age group 40-49 after which the pattern was reversed. Mean total cholesterol in the age range 35-64, after age standardisation, was 229.4 mg/dL. Mean serum total cholesterol was higher in this sample when compared to US population, as well as north and west Indian populations. Thirty-two percent subjects were in the highest risk category with serum cholesterol exceeding 239 mg/dL, while in the US population this fraction constituted only 18 percent. Other risk factors such as high blood pressure, obesity, diabetes, sedentary lifestyle and smoking also had a high prevalence in this population. In this settlement of urban residents in Thiruvananthapuram, serum total cholesterol and low-density lipoprotein cholesterol are high. The causes are likely to be dietary. Combined with the high prevalence of other risk factors such as obesity, hypertension, smoking, diabetes and lack of exercise, this situation demands a preventive programme. PMID- 10820931 TI - Serum fibrinogen and C-reactive protein levels predict major adverse cardiac events in unstable angina. AB - The aim of this study was to ascertain the relationship of raised serum levels of fibrinogen and C-reactive protein at admission with in-hospital major adverse cardiac events in patients with unstable angina and to delineate their angiographic morphology. This single centre, prospective study consisted of 192 patients admitted in an intensive coronary care unit of a large municipal hospital with final diagnosis of unstable angina. The clinical endpoints were: in hospital recurrent angina, new myocardial infarction or cardiac death. Patients with elevated levels of serum fibrinogen and C-reactive protein at admission showed a significantly higher incidence of an in-hospital recurrent major adverse cardiac event (p = 0.001). The mean levels of these markers were also significantly higher in patients with an in-hospital cardiac event as compared to patients with an uneventful hospital stay (p = 0.001). At angiographic evaluation, patients with type B and type C lesions and intracoronary thrombus had significantly higher levels of these markers as compared to patients with type A lesions (p = 0.001). It is concluded that in patients with unstable angina, elevated levels of serum fibrinogen and C-reactive protein at admission indicate an adverse in-hospital outcome and a more complex coronary morphology. The elevated levels of these easily measurable serum markers can therefore be useful in risk stratification of patients with unstable angina. PMID- 10820932 TI - Accelerated infusion of streptokinase in acute myocardial infarction results in better TIMI flow grade in infarct-related artery. AB - Bolus followed by rapid infusion of tissue plasminogen activator results in higher grade of TIMI flow in infarct-related artery as compared to slow infusion. In the present study, an accelerated regimen of streptokinase given over 15 minutes was compared with conventional infusion over one hour in 47 patients presenting within 12 hours of acute myocardial infarction. Forty-seven patients (44 males, 3 females; mean age 54.0 +/- 1.1 years) were randomly allocated to receive 1.5 million units of streptokinase either over 15 minutes (group 1, n = 24) or over one hour (group 2, n = 23) at a mean interval of 5.4 +/- 3.6 hours after onset of symptoms. All the patients received aspirin and intravenous heparin (1000 U/hr) for 96 hours after thrombolysis. Coronary angiography was performed in 43 patients (22 in group 1, 21 in group 2) prior to discharge from the hospital (mean 7 +/- 2.1 days after acute myocardial infarction) and patency of the infarct-related artery and grade of TIMI flow were determined. Infarct related artery was patent (TIMI 2/3 flow) in 19 (86.4%) patients in group 1 as compared to 12 (57.1%) in group 2 (p < 0.05). TIMI grade 3 flow in the infarct related artery was present in 13 (59.1%) in group 1 as compared to 7 (33.3%) in group 2 (p = 0.1). There was no significant difference between group 1 and 2 in time of presentation (mean 5.3 +/- 3.9 hrs vs 5.5 +/- 3.2 hrs), time to needle in hospital (25.6 +/- 11.2 min vs 26.3 +/- 6.2 min), site of infarct (anterior myocardial infarction 12 in group 1 vs 11 in group 2), relief of pain at 90 min (13 vs 12), more than 50 percent reduction of ST elevation at 90 minutes (17 vs 12) and left ventricular ejection fraction (48.8 +/- 9.1% vs 49.8 +/- 16.0%), respectively. Streptokinase was well tolerated in both the groups, although hypotension was more common with the accelerated regimen (5 in group 1 vs 3 in group 2; p = NS). Thus, 'accelerated' streptokinase given over 15 minutes in patients presenting within 12 hours of acute myocardial infarction is well tolerated and results in higher grades of TIMI flow in the infarct-related artery as compared to the "conventional" one-hour infusion regimen. PMID- 10820933 TI - Coronary stenting without pre-dilatation: an observational study. AB - Conventional coronary stenting is done after pre-dilatation of the lesion. The bleeding complications and incidence of subacute stent thrombosis have been reduced significantly by the use of antiplatelet agents and high pressure balloon inflation to ensure complete stent expansion. Elective stenting also can be done without pre-dilatation by "stent alone technique." This approach significantly reduces the procedural cost and ischaemia time, avoiding potential complications such as abrupt vessel closure because of extensive dissection after conventional angioplasty and prior to stent deployment. Eighty patients of stable angina pectoris suitable for coronary angioplasty underwent stenting without pre dilatation. Out of the 100 stents used, 38 were hand-crimped and 62 were pre mounted. The target vessels were left anterior descending artery in 56 percent, right coronary artery in 32 percent and left circumflex in 12 percent. The procedure was successful in 88 percent lesions. In 12 percent stenting could be done only after pre-dilatation. In all these, there was proximal tortuosity and calcification. The fluoroscopy time was 10.2 +/- 4.5 minutes. The average number of balloons used per lesion was 1.08. Stent embolisation occurred in only one patient. There were no major adverse cardiac events in any of the patients. Thus stenting without pre-dilatation is safe. Patients who are eligible for stenting without pre-dilatation are those with stable angina pectoris without fluoroscopically visible calcium or coronary artery tortuosity and with lesions of moderate complexity. PMID- 10820934 TI - Intraoperative transoesophageal echocardiography in aortic valve surgery. AB - From January 1994 to May 1998, 272 patients underwent homograft aortic valve replacement (n = 139), Ross procedure (n = 100) and aortic valve repair (n = 33). Transoesophageal echocardiography was performed intraoperatively before and after cardiopulmonary bypass. Aortic valve morphology, aortic root diameter, pulmonary valve morphology, pulmonary annulus diameter and mitral valve morphology were assessed by two-dimensional imaging. Colour flow mapping was used for assessing severity of aortic regurgitation before and after the procedure. There were no complications related to the procedure. The accuracy of aortic annular diameter measured in the long axis view was confirmed at surgery. The aortic valve morphology was thought suitable for repair and a satisfactory repair was performed in 33 patients as assessed by transoesophageal echocardiography. Post operative transoesophageal echocardiography showed a competent aortic valve in all but four of the remaining 239 patients. Intraoperative transoesophageal echocardiography is easy to learn and provides the surgeon additional information necessary to decide a particular procedure. In addition, intraoperative transoesophageal echocardiography provides accurate assessment of the results of surgery on the table. PMID- 10820936 TI - Profound hypothermic circulatory arrest in management of aortic aneurysms. AB - A total of 15 patients having aneurysms of aorta were operated from June 1997 to December 1998 using deep hypothermic circulatory arrest as a modality of brain protection. There were 12 males and 3 females. The age ranged from 19 years to 74 years and the mean age was 44.9 years. Nine patients had aneurysms of ascending aorta (group I), one had aneurysm of ascending aorta and arch of aorta (group II), four had aneurysm of the distal aortic arch (group III) and one patient had thoracoabdominal aortic aneurysm (group IV). In group I, six patients underwent Bentall procedure, two underwent Wheat procedure and one patient had repair of pseudoaneurysm of ascending aorta. The only patient in group II had his ascending aorta and arch replaced, with reimplantation of left common carotid and innominate artery. In group III, three patients had interposition Gelseal graft and one had repair of the tear in distal aortic arch. The lone patient in group IV had interposition Gelseal graft of thoracoabdominal aorta. The hypothermic circulatory arrest was used in all of them for brain and/or spinal cord protection. Retrograde cerebral perfusion was used in two patients. There were two (13%) operative deaths. One patient died of cerebrovascular accident on eighth post-operative day and second died of inadequate surgical repair. There was one instance of left hemiparesis secondary to an infarct in right frontoparietal region. To conclude, hypothermic circulatory arrest could provide an adequate brain protection for aortic aneurysm surgery. Retrograde cerebral perfusion could be an adjuvant when the anticipated time of hypothermic circulatory arrest is likely to exceed 45 minutes. PMID- 10820935 TI - Management of tetralogy of Fallot with absent pulmonary valve: early and mid-term results of a uniform approach. AB - The operative management of absent pulmonary valve syndrome remains controversial regarding palliative or one-stage correction, the need for pulmonary valve implantation and pulmonary arterioplasty. This retrospective report summarises the experience of a single centre with a view to provide some answers to this controversy. Forty-six consecutive patients including five infants, aged 2 months to 43 years, underwent primary surgical correction during the last 8.5 years. All the patients underwent two-dimensional echocardiography and cardiac catheterisation. Nine patients had mild and 10 moderate pulmonary artery hypertension. Repair consisted of patch closure of the ventricular septal defect and reconstruction of the right ventricular outflow tract. A valve was incorporated in the pulmonary position in 19 patients. Pulmonary arterioplasty was performed only in infants. Overall hospital mortality was 4 out of 46 patients (8.6%). Two out of five infants died accounting for 40 percent mortality. Forty-two survivors were followed up from 4 to 101 months; 40 patients are in functional class I and two in class II. Actuarial survival at 8.5 years was 91 percent. It is concluded that reconstruction of the right ventricular outflow tract with a transannular patch is sufficient in majority of patients. A selective approach to pulmonary valve insertion is recommended in patients with pulmonary hypertension or other anomalies. Pulmonary arterioplasty should be performed as the primary treatment in infants. PMID- 10820937 TI - The "hybrid" approach for revascularisation: direct myocardial revascularisation and coronary angioplasty. AB - We report the cases of two patients where catheter-based laser direct myocardial revascularisation has been coupled with conventional coronary angioplasty at the same sitting using the Biosense left ventricular electromechanically guided laser procedure. In both the cases, the non-revascularizable ischaemic target zone was identified using left ventricular electromechanical mapping signals, and Ho: YAG laser channels were placed at the designated target lesion following successful coronary stenting. The results did not show any procedural complications. This preliminary report suggests the feasibility and safety of this "hybrid" percutaneous approach for myocardial revascularisation. PMID- 10820938 TI - Aortopulmonary fistula following penetrating chest injury. PMID- 10820939 TI - Uhl's anomaly. PMID- 10820940 TI - Genetically transmitted ventricular pre-excitation in a family with hypertrophic cardiomyopathy. PMID- 10820941 TI - Role of infections in the pathogenesis of atherosclerotic coronary artery disease: past, present and future. PMID- 10820942 TI - Nebivolol: a new beta blocker on the horizon. PMID- 10820943 TI - Renoprotection with angiotensin type-1 receptor blockers in hypertension. PMID- 10820944 TI - Intellectual, neuropsychological, and academic functioning in long-term survivors of leukemia. AB - OBJECTIVE: To assess the effects of treatment for acute lymphoblastic leukemia (ALL) on children's cognitive functioning. METHOD: Participants were long-term survivors of ALL treated with cranial irradiation and central nervous system (CNS) chemotherapy (n = 20), or CNS chemotherapy only (n = 21), healthy children (n = 21), and children with chronic asthma (n = 21). The groups were compared on measures of intellectual, neuropsychological, and academic functioning. RESULTS: CNS chemotherapy, with and without cranial irradiation, was associated with significantly lower levels of intellectual and academic functioning. Children with chronic asthma obtained lower scores than healthy controls, but these differences were not significant. Tests of neuropsychological functioning did not consistently separate the groups. CONCLUSIONS: CNS chemotherapy and, to a lesser extent, chronic illness both contribute to the poorer performance of long-term survivors of ALL on measures of intellectual and academic functioning. PMID- 10820945 TI - The parent-physician relationship in pediatric asthma care. AB - OBJECTIVE: To examine the feasibility of using a behavioral coding system, the Five Minute Speech Sample (FMSS), to estimate the prevalence of relationship difficulties between parents and physicians in a pediatric asthma care setting. METHOD: The FMSS was administered to 20 parents of children with severe, chronic asthma and to the physicians they worked with during a brief day program admission. RESULTS: Rates of relationship difficulties between parents and physicians ranged from 15% to 40%, comparable to those previously identified in patient-therapist relationships in psychiatric populations. CONCLUSIONS: Although further information is needed about validity and reliability, the FMSS appears to provide clinically relevant information about the parent-physician relationship in the context of pediatric chronic illness. PMID- 10820946 TI - Predictors of distress in parents of children with cancer: a prospective study. AB - OBJECTIVE: To investigate levels of psychological distress in parents of children with cancer and relationships between distress and measures of illness variables, appraisal, psychosocial resources and coping strategies. METHODS: Questionnaires were completed by 68 mothers and 58 fathers at 6 (Time 1) and 18 (Time 2) months post-diagnosis. RESULTS: High levels of distress for 51% of mothers and around 40% of fathers were apparent at both time points, with little change over time. For mothers, their appraisal of the strain of the illness, and their own ability to deal with it, and family cohesion were predictive of distress, both concurrently and prospectively, and, at Time 1, greater use of self-directed coping strategies was related to higher levels of distress. For fathers, risk factors of employment problems (Time 1) and the number of the child's hospital admissions (Time 2) were significant, along with appraisal and family cohesion. CONCLUSIONS: Continuation of high levels of distress over time points to the importance of identification of those at risk at an early stage and provision of ongoing support. Implications for such support are discussed. PMID- 10820947 TI - Psychosocial adjustment in children with kidney disease. AB - OBJECTIVE: To examine family environment, levels of parenting stress, and child behavior problems in children with one of three kidney diseases compared to healthy children and to examine predictors of psychological distress in the full sample. METHOD: Parents of children with steroid sensitive nephrotic syndrome, chronic renal insufficiency, or kidney transplant (n = 41) were compared to 34 healthy children of similar demographic characteristics. RESULTS: Mean scores on family functioning, parenting stress, and child behavior were within normal limits. Family environment variables significantly predicted child behavior and parenting stress for parents of ill and healthy children. Qualitative responses provided insight into developmentally specific stressors and intervention needs in the illness groups. CONCLUSIONS: These data indicate that long-term survivors of kidney disease function similarly to demographically matched peers and that the family environment may buffer stress caused by illness. Specific concerns raised by parents in the kidney disease groups indicate the need to appropriately assess and intervene with this understudied population. PMID- 10820948 TI - Identity in adolescent survivors of childhood cancer. AB - OBJECTIVE: To investigate identify formation among adolescent survivors of childhood cancer. Family functioning, perceived emotional support from family and peers, life stress, and anxiety produced by the cancer experience also were examined as they influenced identity development. METHOD: Participants were 52 adolescent survivors and their mothers recruited from a medical center and 42 healthy adolescent counterparts and their mothers recruited from the community. RESULTS: A greater frequency of survivors than their healthy peers was found within the foreclosed identity status. Factors associated with the foreclosed identity status included the cancer diagnosis, symptoms of posttraumatic stress disorder (PTSD), and family functioning characterized by greater levels of conflict. CONCLUSIONS: Data were interpreted to suggest that the foreclosed identity status may serve a protective function in assisting survivors to cope with the stressors of the cancer experience. PMID- 10820949 TI - Brief clinical report: procedural pain and anxiety management with mother and sibling as co-therapists. AB - OBJECTIVE: To describe a multidimensional family-focused intervention for pain and anxiety management during port-access with a Spanish-speaking preschool child being treated for acute lymphoblastic leukemia, using the patient's mother and one sibling as co-therapists. METHODS: Assessment procedures included interviews with mother, sibling, and primary nurse and behavioral observations during port access, and during a medical examination. Intervention consisted of two training and two "in-vivo" sessions, using behavioral and cognitive behavioral techniques. RESULTS: Physical resistance was eliminated and anxiety responses reduced during port-access sessions. Maternal anxiety and sibling distress were also reduced. CONCLUSIONS: This case study illustrates an empirically supported child and family focused in the clinical care context and highlights cultural factors that may influence the success of such interventions. PMID- 10820950 TI - Factors affecting the outcome of surgical treatment of acromegaly. AB - PURPOSE: To assess the results of and factors associated with the outcome of surgery for acromegaly. METHODS: We retrospectively examined the medical records of acromegalic patients who underwent trans-sphenoidal adenomectomy at our hospital during the period of January 1991 through August 1997. Preoperative evaluations included measurement of basal serum growth hormone (GH), insulin-like growth factor-I (IGF-I), prolactin (PRL), GH response to oral glucose, and GH and PRL response to bromocriptine, as well as pituitary magnetic resonance (MR) imaging. Postoperative evaluations included measurement of basal serum GH and IGF I concentrations, and pituitary MR imaging. RESULTS: Thirty patients (14 men) with a mean age of 38 years were included. The mean follow-up period was 50 months (range, 15-90 mo). Ten of the 30 patients (33%) had early postoperative (1 mo after surgery) GH levels of less than 5 ng/mL. Twenty patients (67%) had final postoperative (last follow-up, 15-90 mo after surgery) GH levels of less than 5 ng/mL. Preoperative GH levels were positively correlated with early postoperative GH levels (r = 0.458, p = 0.011) and final postoperative GH levels (r = 0.479, p = 0.007). Early postoperative GH levels were also positively correlated with final postoperative GH levels (r = 0.595, p = 0.001). Tumor grade and stage were not significantly correlated with early or final postoperative GH levels. Thirteen of 21 patients (62%) who had postoperative MR imaging follow-up had residual tumor. There was no surgical mortality. CONCLUSIONS: These results highlight that acromegaly is not easily treated with surgery alone. The preoperative GH level was associated with the surgical outcome. PMID- 10820951 TI - Toxic megacolon secondary to infective colitis in children. AB - BACKGROUND AND PURPOSE: Toxic megacolon is a fulminating and potentially lethal complication of severe colitis. Toxic megacolon secondary to infective colitis in children is rare. We analyzed the clinical course, pathology, treatment, and outcome of toxic megacolon secondary to infective colitis in children. METHODS: The medical records of all 20 children treated for infective colitis complicated with toxic megacolon during a 12-month (October 1997-October 1998) period were retrospectively reviewed. RESULTS: There were 10 boys and 10 girls, with a mean (+/- standard deviation, SD) age of 26.2 +/- 12.9 months (range, 6-57 mo). With an initial presentation of nonspecific gastroenteritis syndrome lasting several days, the disease progressed rapidly. In the acute stage, most patients developed toxic signs such as mental change, ranging from irritability to stupor (20, 100%), fever (19, 95%), tachycardia (20, 100%), abdominal distension (20, 100%), and abnormal stool pattern (19, 95%). Initial investigations revealed anemia (11, 55%), leukocytosis (11, 55%), and elevated levels of C-reactive protein ranging from 25.0 mg/L to 483.0 mg/L with a mean +/- SD of 185.7 +/- 129.1 mg/L (normal range, < 8 mg/L) (20, 100%). Salmonella enteritidis (12 patients, 60%) and Clostridium difficile (1, 5%) were isolated from stool samples in some cases. Plain abdominal x-rays revealed severe colonic dilatation. Prolonged hospitalization (mean, 33.6 d) and intensive therapy including a combination of broad-spectrum antibiotics, physical decompression, and total parenteral nutrition were necessary. Three patients (15%) underwent surgical management; the pathologic findings in these patients demonstrated severe transmural inflammation. We believe that bacterial and/or endotoxin translocation played an important role in gut failure. Three patients (15%) in the study died. CONCLUSION: Toxic megacolon in infective colitis is a fulminating illness that has a high mortality rate. The disease course can be divided into three stages: the acute toxic stage, the gut failure stage, and the convalescence or deterioration stage. Early diagnosis and aggressive management are important. PMID- 10820952 TI - Safety and effectiveness of percutaneous fluoroscopic gastrostomy in cancer patients. AB - PURPOSE: To evaluate the safety and effectiveness of percutaneous fluoroscopic gastrostomy (PFG) in cancer patients. METHODS: We retrospectively analyzed the success rate, complication rate, and patient outcomes of PFG performed during a 15-month period in our hospital. The Cope type 10-French and 12-French self retaining catheters were used for gastrostomy. RESULTS: Of the 113 PFG procedures requested during the study period, 112 (99.1%) were performed successfully; one procedure was cancelled because the nasogastric tube could not be passed through the obstructed esophagus. Thirty-day follow-ups were obtained for 92.0% of the procedures. Of the patients with at least 30 days of follow-up, eight (7.8%) died, but only two deaths (1.9%) were procedure-related. Major complications, including peritonitis and severe wound infection, occurred after eight procedures (7.8%). Minor complications, including superficial wound infection, tube fracture, leakage, severe pain, mild hemorrhage, and tube migration, occurred after 23 procedures (22.3%). The severity of pneumoperitoneum was significantly associated with the frequency of complications (p < 0.05). After placement of the gastrostomy, 68% of the patients maintained or increased their initial body weight; 28% lost 10% or less of their body weight; and 4% lost more than 10% of their body weight. CONCLUSIONS: PFG appears to be effective and relatively safe for long-term nutritional support in cancer patients. PMID- 10820953 TI - Selective inhibition of inducible nitric oxide in ischemia-reperfusion of rat small intestine. AB - PURPOSE: We investigated the role of constitutive and inducible nitric oxide (NO) synthases in intestinal ischemia-reperfusion (I/R) injury by observing the alterations in hemodynamics and intestinal microcirculation in response to I/R in rats, with or without inhibitors of NO synthases. METHODS: Adult male Sprague Dawley rats (n = 9/group) received a standard I/R procedure alone: I/R plus intravenous administration of aminoguanidine (an inhibitor of inducible NO synthase); I/R plus L-NAME (NG-nitro-L-arginine methyl ester, an inhibitor of constitutive and inducible NO synthase); IR + L-Arg (L-arginine, an NO precursor); or a sham operation plus the vehicle. The I/R procedure was performed by clamping the perfusion vessels of a segment of the terminal ileum, and medication was administered intravenously before and after intestinal ischemia. The intestinal perfusion and leukocyte-endothelial interactions were evaluated with in vivo microscopy and laser Doppler flowmetry. Surface expression of CD11b (an adhesion molecule) of circulating granulocytes was measured with flow cytometry. RESULTS: Intestinal I/R produced circulatory alterations, intestinal microcirculatory derangement, energy depletion, and lipid peroxidation. Aminoguanidine significantly attenuated the reperfusion-related depression of mean arterial pressure (MAP), the decrease in intestinal perfusion index, the decrease in tissue ATP preservation, the increase in tissue malondialdehyde (MDA) level, and the expression of CD11b of circulating granulocytes. Administration of L-NAME had only minor and transient effects on reperfusion-related changes of MAP, intestinal flux, numbers of adherent leukocytes, and CD11b expression, but had some protective effects on tissue MDA and adenosine triphosphate levels and flow velocity. L-Arg further decreased the MAP but did not affect reperfusion related variables. CONCLUSIONS: Our results show that the selective inhibition of inducible NO synthase by aminoguanidine attenuates the hemodynamic and microcirculatory derangement that results from intestinal I/R. PMID- 10820954 TI - Diffuse toxoplasmic retinochoroiditis as the initial manifestation of acquired immunodeficiency syndrome. AB - Toxoplasmic retinochoroiditis is an important opportunistic retinal infection in human immunodeficiency virus (HIV)-infected patients. It may present as diffuse necrotizing retinochoroiditis instead of a focal lesion and may be the initial manifestation of HIV infection. A 50-year-old heterosexual man presented with blurred vision in his left eye of 3 months' duration. Fundus examination revealed diffuse necrotizing retinochoroiditis, mainly at the posterior pole, with marked vitritis in the left eye. Serologic studies and aqueous fluid antibody titers indicated recent toxoplasmic infection. Positive enzyme immunoassays (EIA) and Western blot tests proved HIV infection. The retinochoroiditis and vitritis improved after an antitoxoplasmic regimen with trimethoprim-sulfamethoxazole (TMP SMX). Nonetheless, toxoplasmic encephalitis developed 6 months after the onset of ocular toxoplasmosis and responded well to TMP-SMX. This is the first case of toxoplasmic retinochoroiditis as the initial manifestation of AIDS reported in Taiwan. We suggest that Toxoplasma infection should be included in the differential diagnosis of diffuse necrotizing retinochoroiditis and vitritis. We also recommend that adults with newly diagnosed ocular toxoplasmosis be screened for HIV infection. PMID- 10820955 TI - Pertussis seroepidemiology in Taipei. AB - PURPOSE: We evaluated the seroprevalence of pertussis among apparently healthy Taipei residents. METHODS: From January 1992 through December 1994, we recruited subjects from a well baby clinic (children < 3 years), kindergartens, primary, and secondary schools and colleges (3-20 years), a health exam clinic (> 20 years), and obstetric clinics (pregnant women). Subjects were stratified into 12 groups according to age and pregnant women were considered separately. The serum antibody titers against filamentous hemagglutinin (FHA) and pertussis toxin (PT) were measured by enzyme-linked immunosorbent assay. RESULTS: The anti-PT and anti FHA titers were elevated in the 4 to 6-month and 19 to 20-month age groups, coinciding with regular pertussis vaccinations. The anti-PT titers rose again in the 3 to 4-year age group, reflecting a higher prevalence of natural pertussis infection. The anti-PT titers gradually decreased among older age groups, although a peak occurred in the 11 to 15-year age group. The sequential changes in anti-FHA titers followed those of anti-Pt titers in the younger age groups, but the anti-FHA titers were persistently elevated beyond 15 years of age. The antibody levels were similar in the two sexes, except that anti-PT titers were higher in males of 19 to 20 months and 21 to 30 years of age. Anti-PT titers were equivalent between neonates (0-2 months) and pregnant women, but anti-FHA titers were much lower in neonates. CONCLUSIONS: The seroprevalence patterns in this study indicate that young children, adolescents, and even adults remain at risk of pertussis, despite the current immunization program. Booster vaccinations after completion of the current four-dose immunization schedule, possibly continuing into adolescence, should be considered to block the transmission of infection. PMID- 10820956 TI - Expression of p53 protein in oral submucous fibrosis, oral epithelial hyperkeratosis, and oral epithelial dysplasia. AB - BACKGROUND AND PURPOSE: In our previous study, positive p53 staining was observed in 47 of 81 (58%) cases of oral squamous cell carcinoma associated with areca quid (AQ) chewing and cigarette smoking. This study looked for expression of p53 protein in premalignant oral lesions in patients who chewed AQ or smoked cigarettes, or both. METHODS: Expression of p53 protein was examined in histologic sections of oral submucous fibrosis (OSF, n = 50), epithelial hyperkeratosis (EH, n = 10), epithelial dysplasia (ED, n = 10), and normal oral mucosa (NOM, n = 10) with antibodies against p53 protein using an immunoperoxidase technique. RESULTS: Positive p53 staining was observed in 30 (60%) OSF specimens, four (40%) EH specimens, seven (70%) ED specimens, and none of the NOM specimens. Only four (8%) of the OSF specimens and none of the EH specimens had more than 25% p53-positive keratinocytes. However, in four (40%) of the ED specimens, more than 50% of the keratinocytes were p53-positive. The degree of p53 staining increased with the morphologic transformation of normal appearing epithelial cells into dysplastic epithelial cells. There was no significant correlation between expression of p53 in OSF epithelium and the clinicohistologic parameters of patients with OSF. CONCLUSIONS: These results demonstrate that p53 is often present in precancerous lesions of patients who chew AQ and smoke cigarettes. We suggest that p53 may play a role in dysplastic cell transformation in premalignant oral lesions. PMID- 10820957 TI - Quality assurance in clinical laboratories in Taiwan. AB - BACKGROUND AND PURPOSE: In 1994, the Taiwan National Health Administration assigned the execution of a quality assurance (QA) survey program to the Association of Laboratory Medicine. The purpose of this program was to investigate the quality of clinical laboratory assessments and to promote QA in the fields of clinical microscopy, hematology, chemistry, microbiology, serology, and blood banking. We report the findings of QA surveys conducted in 1998 and the effect of voluntary training on improvement of clinical laboratory testing. METHODS: A total of 1,008 clinical laboratories were included in the program in 1998. Proficiency testing (PT) was performed to evaluate various laboratory tests. Continuing education programs were conducted and experts visited laboratories that sought guidance before the PT was conducted. The full mark was set at a score of 100 for each PT scheme. The criterion for acceptability of PT results was set at a score of 80 or more. RESULTS: The rates of acceptable results were 82.4% (607/736) for hematology, 57.4% (267/465) for blood banking, 69.3% (561/810) for chemistry, and 80.1% (321/401) for microbiology. The rates of acceptable microscopy results were 90.9% (509/560) for urine chemical tests and 84.6% (610/721) for others. The rates of acceptable serology tests were 73.3% (384/524) for hepatitis and 85.6% (441/515) for syphilis. The rates of acceptable performance differed significantly among clinical laboratories with different rankings: clinical laboratories at institutions classified below the level of district hospital showed comparatively poor performance. Laboratories that received guidance showed significant improvement in performance from 1997 to 1998. CONCLUSIONS: A QA program is urgently needed in Taiwan to improve laboratory performance. PMID- 10820958 TI - Idiopathic multicentric osteolysis with nephropathy. AB - Idiopathic multicentric osteolysis is a rare syndrome that manifests with progressive loss of carpal and tarsal bones in childhood. Affected children have arthritic-like episodes, followed by progressive deformities, radiographic osteolytic changes, and variable degrees of disability. A rare form of this disease (type III, sporadic) is associated with serious nephropathy. We present the first reported case of type III idiopathic multicentric osteolysis in a Chinese woman. The patient, a 34-year-old woman with normal mental development and no family history of bone or kidney disease, presented with a 4-day history of nausea and vomiting. She had shortening and swelling of the hands, which had occurred in childhood and persisted at the time of admission. X-ray studies showed disappearance of the carpal bones, and multiple osseous erosions of the tarsal bones. Hypertension, severe azotemia, and metabolic acidosis were also noted. Advanced renal disease was documented after a series of investigations, including renal biopsy. She is now dialysis-dependent. This case illustrates the importance of early diagnosis and management of idiopathic multicentric osteolysis with nephropathy. PMID- 10820959 TI - Cyclosporine-induced encephalopathy in a patient with relapsed acute myeloid leukemia treated with unrelated allogeneic bone marrow transplantation. AB - Cyclosporine (CSP) is the most frequently used immunosuppressive agent for prevention of graft versus host disease (GVHD) in allogeneic bone marrow transplantation (BMT). Some adverse effects such as hepatic and renal toxicity have been frequently encountered, but central nervous system (CNS) toxicity caused by CSP is rare. We report an adult male patient with acute myeloid leukemia who developed CSP-induced encephalopathy under treatment for allogeneic BMT from an unrelated donor. Methotrexate and CSP were used for GVHD prophylaxis. Leukocyte and platelet engraftment were successfully achieved on days 21 and 24 after BMT, respectively. Abrupt onset of mental confusion and disorientation occurred on day 25, followed by a generalized tonic clonic seizure and consciousness disturbance. The whole blood CSP level was 160.65 ng/mL. Magnetic resonance (MR) imaging revealed high signal intensities in the bilateral occipital lobes with predominant involvement of the cortical areas. The patient recovered from the CNS toxicity, but with slight memory impairment, 6 days after CSP was discontinued. When patients receiving CSP treatment for allogeneic BMT develop mental confusion, consciousness disturbance, or seizure, CSP-induced CNS toxicity should be taken into consideration. PMID- 10820960 TI - Swyer-James syndrome complicated by lung abscess. AB - Swyer-James syndrome, a rare disease with unilateral hyperlucent lung due to bronchiolitis obliterans and pulmonary artery hypoplasia, generally develops after lower respiratory tract infection during early childhood. Invasive procedures, including bronchoscopy and angiography, are often necessary for a definitive diagnosis. We report a 17-year-old man admitted because of cystic bronchiectasis complicated by lung abscess. Chest roentgenography showed the typical findings of Swyer-James syndrome. Noninvasive magnetic resonance angiography was used to confirm hypoplasia of the right pulmonary artery. The patient received antibiotic therapy, underwent a right lower lobectomy for the lung abscess, and recovered. PMID- 10820961 TI - Use of extracorporeal membrane oxygenation to rescue a child with acute respiratory distress syndrome. AB - Acute respiratory distress syndrome (ARDS) carries a high mortality of about 60%. The results of conventional treatments for ARDS are poor. We report the use of extracorporeal membrane oxygenation (ECMO) to rescue a child with ARDS. The patient, a 7-year-old boy, underwent a Ross procedure and mitral valvuloplasty because of severe aortic and mild mitral regurgitation. ARDS due to massive transfusion and prolonged cardiopulmonary bypass developed in the early postoperative period. Hypoxemia persisted despite conventional treatments, including pressure-controlled ventilation and high-frequency ventilation. Finally, venovenous ECMO was used to rescue the patient. With ECMO support, gas exchange was well maintained with a lower ventilator setting, and ventilator induced lung injuries were avoided. ECMO was used for 183 hours, at which point the boy was weaned without complications. His recovery was uneventful. At the latest follow-up, 6 months after the operation, he was in New York Heart Association function class I and had no complaints of lung disease. This case suggests that venovenous ECMO can be a rescue method for patients with ARDS that is refractory to conventional treatments. PMID- 10820962 TI - Partial left ventriculectomy as a biologic bridge to heart transplantation. AB - Heart failure refractory to medical treatment is consuming an increasingly large proportion of health care resources. Partial left ventriculectomy has recently been used to treat patients with end-stage dilated cardiomyopathy. A 45-year-old man in end-stage heart failure had progressive exertional dyspnea for 3 years. Cardiac catheterization showed dilated cardiomyopathy with severe pulmonary hypertension and severely impaired left ventricular function. After partial left ventriculectomy, cardiac output increased from 2.11 L/min to 5.0 L/min. The left ventricular ejection fraction measured by radionuclide angiography increased from 13% preoperatively to 28% 1 month after the operation. The patient was discharged and monitored at the outpatient clinic. He had an exercise capacity of NYHA functional class II. However, he received heart transplantation 10 months after the partial left ventriculectomy because of recurrent heart failure. Partial left ventriculectomy improves heart function and may be used as a bridge to heart transplantation as an alternative to ventricular assist devices. PMID- 10820963 TI - Persistent myocardial ischemia after termination of dipyridamole-induced ventricular tachycardia by intravenous aminophylline: scintigraphic demonstration. AB - Ventricular tachycardia is a rare complication of dipyridamole stress testing. We present a case in which dipyridamole induced symptomatic ventricular tachycardia. The patient, a 41-year-old man with a history of chest discomfort on exertion for 1 year, underwent dipyridamole thallium imaging. Sustained ventricular tachycardia occurred 1 minute and 40 seconds after completion of the dipyridamole infusion. Intravenous aminophylline (125 mg) was given immediately and the tachycardia was terminated. Two minutes later, thallium-201 was injected and subsequent myocardial imaging showed reversible perfusion defects in the inferior wall, septum, and apex. Coronary angiography revealed three-vessel disease. This case discloses that reversible perfusion defects can still be demonstrated on thallium scan in spite of injection of aminophylline, an antagonist of dipyridamole, before thallium administration. PMID- 10820964 TI - Treatment of chronic pain by spinal cord stimulation. AB - Chronic pain after injury of the nervous system is difficult to treat. This report describes our experience in the treatment of chronic pain by spinal cord stimulation (SCS) in four patients (three men and one woman, with ages ranging from 61 to 73 yr). One patient had chronic intractable pain due to a spinal cord injury, one had post-herpetic neuralgia, one had failed back surgery syndrome, and one had brachial plexus and spinal cord injuries. A permanent spinal cord stimulator was implanted in the patient with spinal cord injuries. In the other three patients, a permanent spinal cord stimulator was implanted after a successful trial stimulation with temporarily implanted electrodes. After 19 to 25 months (mean, 21 mo) of follow-up, three patients had satisfactory improvement of pain and one patient had temporary pain relief, but pain recurred two months after implantation. One patient had recurrence of pain after migration of an electrode, and the pain-relieving effects of SCS returned after repositioning the electrode. According to this preliminary experience, SCS is effective for pain reduction in selected patients with chronic pain. Long-term follow-up is mandatory to evaluate the benefits and complications of SCS for relief of chronic pain. PMID- 10820965 TI - Was typhoid inoculation safe and effective during World War I? Debates within German military medicine. PMID- 10820966 TI - Nettleship, Pearson and Bateson: the biometric-Mendelian debate in a medical context. PMID- 10820967 TI - The Duchenne de Boulogne-Meryon controversy and pseudohypertrophic muscular dystrophy. PMID- 10820968 TI - Poverty and child abuse: suffer the children. PMID- 10820969 TI - Addressing the mental health needs of the poor. PMID- 10820971 TI - The positive economic impact of health care for the homeless. PMID- 10820970 TI - Characteristics of persons with AIDS in rural and urban Georgia: use of alcohol and other drugs, history of sexually transmitted diseases and tuberculosis, high risk sexual behaviors, and problems obtaining social services. PMID- 10820972 TI - Making room at the table: pushing aside poverty and hunger. PMID- 10820973 TI - Giving Georgia's babies a healthy beginning. PMID- 10820974 TI - Health status and socio-economic status in Georgia: a quality of life perspective. PMID- 10820975 TI - A call to action: growing health insurance problem threatens vitality of Georgia hospitals. PMID- 10820976 TI - State must close health insurance gap. PMID- 10820977 TI - Poverty, race and prevention. PMID- 10820978 TI - The challenges of providing home health care to the indigent population. PMID- 10820979 TI - Colon cancer discovery and the American Cancer Society neglect of single contrast barium enema. PMID- 10820980 TI - Maxillomandibular advancement surgery for obstructive sleep apnea. PMID- 10820981 TI - Preparing for HIPAA. PMID- 10820982 TI - Are we any closer to being able to measure leprosy exposure? PMID- 10820983 TI - Women with leprosy. A woman with leprosy is in double jeopardy. PMID- 10820985 TI - Which physical signs help most in the diagnosis of leprosy? A proposal based on experience in the AMFES project, ALERT, Ethiopia. AB - As integration of leprosy control programmes proceeds, general health staff will have responsibility for the diagnosis of most new cases of leprosy. The training required by these workers has not yet been set out in detail. In this paper the criteria for making the diagnosis of leprosy in the AMFES cohort of 594 new cases are examined. Since this study does not include details of suspects in whom leprosy was excluded on clinical grounds, true sensitivity and specificity values cannot be calculated, but the positive predictive value of the diagnostic criteria can be measured. Sensory loss in a typical skin patch is the most important sign of early leprosy, but was not present in 132 (49%) of the 268 cases with a positive skin smear. Thickening of the ulnar nerve is a valuable sign of leprosy in Ethiopia. It can be taught to health workers, who can practise by examining their own ulnar nerves. It is more likely to be present than nerve function impairment and is particularly important when skin smears are difficult to do or are unreliable. We recommend that five basic signs are used, the presence of any two being diagnostic of leprosy: Skin lesion(s) consistent with leprosy. Loss of sensation in such a lesion. Thickening of either ulnar nerve. Loss of sensation in the palm of the hand or the sole of the foot. The presence of acid-fast bacilli in skin smears. Exact policies for the diagnosis of leprosy should be worked out and validated for each national programme. PMID- 10820984 TI - Incidence rates of acute nerve function impairment in leprosy: a prospective cohort analysis after 24 months (The Bangladesh Acute Nerve Damage Study). AB - In this paper, the incidence rates and cumulative incidence of nerve function impairment (NFI) and leprosy reactions over 24 months follow-up of the prospective cohort of 2664 new leprosy cases are presented. Graphs showing the cumulative incidence of NFI relative to time since registration are presented. Hazard ratios (HRs) for the development of NFI for four variables are given. The majority of patients who developed NFI after registration did so in the first year (67% of multibacillary (MB) patients, and 91% of paucibacillary (PB) patients who developed NFI). Thirty-three percent of all MB patients who developed NFI after registration did so in the second year of follow-up. No PB patients developed NFI for the first time in the last 6 months of follow-up. However, seven NFI events occurred amongst PB patients in that period, amongst those who had already had one NFI event. The incidence rate (IR) of NFI amongst MB patients was 24/100 person-years at risk (PYAR), and amongst PB patients was 1.3/100 PYAR. The HR for the development of NFI amongst MB patients compared with PB patients was 16 using univariate analysis. Amongst patients who had long standing NFI present at registration, the IR was 27/100 PYAR compared with 1.7/100 PYAR amongst those who did not have long-standing NFI. The HR for developing acute NFI amongst those with long-standing NFI present at registration compared with those without was 14 using univariate analysis. When multivariate regression analysis is applied, the apparently significant univariate HRs for sex and age disappeared. The resultant multivariate HR for leprosy group is 8.8, and 6.1 for the presence/absence of long-standing NFI at registration. In all, 142/166 (86%) of all new NFI events were silent, underlining the need for regular nerve function testing. IRs are presented for the four 6-month periods of the 24 month follow-up. They show a clear stepwise reduction over the total period. The IRs amongst MB patients and those with long-standing NFI present at registration are very high at 34 and 41/100 PYAR, respectively, for the first 6 months of follow-up. Even during the final 6-month period, the IR is maintained at a moderately high level (18 and 15/100 PYAR, respectively). PMID- 10820986 TI - A 6 week quadruple drug regimen for the treatment of multibacillary leprosy. PMID- 10820987 TI - Computerization of leprosy records: national leprosy recording and reporting system in China. AB - This paper describes the national system of leprosy recording and reporting in China and the computerization of records. The system was designed for data collection at local level and data entry by optically scanned or manual mode as well as for sophisticated data analysis. The major functions include data entry, data check, sum-up, maintenance, communication, inquiry, statistics, graph and print. A total of 17 options for epidemiological and clinical data analysis are available. Through the implementation for about 10 years, the system has gained widespread acceptance. This acceptance would facilitate introduction of computer analysis to other leprosy projects and other disease control programs in China. Up to 1998, a database of more than 740,000 records covering all the leprosy patients detected since 1949 had been established by this system. PMID- 10820989 TI - A new face for an old disease: some reflections on the role of the media in Nepal's first National Leprosy Elimination Campaign. PMID- 10820988 TI - A survey on knowledge and skills in the early diagnosis of leprosy in general health services at different levels in Shandong Province, The People's Republic of China. AB - In the late phase of a leprosy control programme, problems arise with regard to the early detection and treatment of a small number of new incident cases. We describe a study in the province of Shandong, People's Republic of China, on the knowledge and skills regarding leprosy of general health service staff, including rural doctors, paramedical doctors at township level, doctors from county general and provincial hospitals and dermatologists. The results showed that there is a continuing need for suitable training programmes for medical staff in the general health services. Most dermatologists had good levels of knowledge and skills and more than 80% of new cases have been diagnosed in skin clinics in this province since 1990. Their participation in early diagnosis and training of staff should be strengthened. PMID- 10820990 TI - Recent trends in leprosy in a large district of West Bengal, India, revealed by a modified leprosy elimination campaign (MLEC), 1998. AB - A Modified Leprosy Elimination Campaign (MLEC) in September 1998 in the District of Midnapore, West Bengal, covered a population of 8.1 million people and detected 8181 new cases. Available data from 7328 cases were studied to observe the trend for leprosy in this area. Data are presented on sex and age distribution, classification and the proportions of multibacillary (MB), paucibacillary (PB) and single skin lesion (SSL) cases discovered in a period of only 8 days. The large numbers of people examined in this district and the high total of new cases revealed are in keeping with experience in other parts of the State and in other parts of India. However, many cases were found in endemic areas and these will receive special attention in a second MLEC, planned for January 2000. PMID- 10820991 TI - Leprosy elimination campaign, Amazonas-Brazil 1997. AB - A leprosy elimination campaign (LEC) was carried out in 15 endemic areas of Amazonas State, Brazil, in 1997. The LEC concentrated effort to detect leprosy cases during a multi-vaccination national campaign for serious public health problems other than leprosy, such as polio, diphtheria, hepatitis, measles, etc. The national campaign involved intensive population mobilization, giving a valuable opportunity to examine people for leprosy. The LEC personnel included 2964 individuals (municipal and state health workers and community volunteers), distributed in 688 health units and 53 reference health centres. As a result of the LEC, 74,814 person-to-person communications in the community were given; 10,297 clinical skin examinations were conducted, and 40 new leprosy cases were detected on the day of the campaign in urban areas of the municipalities. This total was low, compared to results in other states of Brazil, possibly due to the development of health education activities and regular community services in the state of Amazonas since 1987 and to the early implementation of WHO multiple drug therapy (MDT) from 1982 onwards. Despite the fact that the LEC was carried out only in the urban areas of the municipalities, the finding of no cases of leprosy in 7 out of 15 of them was surprising and may indicate that the prevalence of hidden cases of leprosy is not all that high, at least in these areas of the Amazonas State. PMID- 10820992 TI - Genital nodules and testicular hydrocele in a case of relapsed lepromatous leprosy. PMID- 10820993 TI - Radio as a means to enhance early case finding in leprosy. PMID- 10820994 TI - Sensory testing using neurothesiometry. PMID- 10820995 TI - Grading impairment in leprosy. PMID- 10820996 TI - Third International Conference on the Elimination of Leprosy. PMID- 10820997 TI - 21st Biennial Conference of Indian Association of Leprologists (IAL) held at Chandigarh, India. PMID- 10820998 TI - 'The past & present of leprosy': international congresses on the evolution and paleoepidemiology of infectious diseases, Bradford University UK, July 1999. PMID- 10820999 TI - 'Cytotoxic T-lymphocytes against malaria and tuberculosis: from natural immunity to vaccine design' PMID- 10821000 TI - Tuberculosis. An interdisciplinary perspective PMID- 10821001 TI - Leishmania and HIV co-infection. PMID- 10821002 TI - WHO: diagnostic discovery operations, with emphasis on TB. PMID- 10821003 TI - WHO: vaccine research PMID- 10821004 TI - Improving malaria information support for health professionals in southern Africa: 'Red Malaria Reference Initiative'. PMID- 10821005 TI - Global alliance for leprosy elimination. PMID- 10821006 TI - First annual April (fool's) commentary. Cognitive sinks: the black holes of neuropsychology: a brief overview. PMID- 10821007 TI - October 26, 1977: a day to cherish. PMID- 10821008 TI - A surgical approach to low back pain. PMID- 10821009 TI - A non-surgical approach to low back pain. PMID- 10821010 TI - Physicians in Rhode Island. PMID- 10821011 TI - Bone metastases. PMID- 10821012 TI - Spinal surgery for severe scoliosis in Parkinson's disease. PMID- 10821013 TI - A resource for the primary care physician. PMID- 10821014 TI - The thiazolidinediones or "glitazones" a treatment option for type 2 diabetes mellitus. AB - The glitazones offer a promising alternative to patients not currently able to achieve target gylcemic control with current therapy. Glitazones have a synergistic effect when combined with sulfonlyureas and metformin and may have a favorable effect on the dyslipidemia experienced by patients with type 2 diabetes. The main concern with these agents is safety due to the hepatocellular injury experienced by several patients taking troglitazone. Although elevations in serum alanine amino transferase (ALT) concentrations more than three times normal occurred in 1.9% of patients on troglitazone in clinical trials, liver toxicity was not truly associated with this drug until post-marketing use by more than 600,000 people in the US. The onset of elevated ALT is typically delayed. In the clinical trials only one patient experienced an elevation in the first month, the majority occurred between the third and seventh month. The glitazones should be avoided in patients with liver disease or a history of alcohol abuse. The mechanism of action for troglitazone-induced liver disease is unknown; consequently, identifying a particular subset of people as being at an increased risk for developing liver failure is difficult. Therefore, monitoring liver function is of critical importance. The newer agents rosiglitazone and pioglitazone may provide a safer alternative, however this question will remain unanswered until clinicians have access to substantially more post marketing surveillance data. The two recent cases of hepatocellular injury associated with rosiglitazone therapy further supports a cautious approach to utilization of these agents. PMID- 10821015 TI - Progress in the early identification of breast cancer, Rhode Island, 1987-1998. PMID- 10821016 TI - Bone mineral density assessment. PMID- 10821017 TI - Disclosure--is the worm turning, at last? PMID- 10821018 TI - The CPIA in practice--the legal profession's experience. PMID- 10821019 TI - Home office/government perceptions, past and present. Proposals for future strategy. PMID- 10821020 TI - Arrest in Thunder-Ten-Tronckh. PMID- 10821021 TI - The prosecuting authority's role. Making the Criminal Procedure and Investigations Act work to facilitate fair trials and just verdicts. PMID- 10821022 TI - The CPIA disclosure regime; PII and third party disclosure, the defence perspective. PMID- 10821023 TI - Prosecution disclosure in criminal cases: the European Convention on Human Rights and the Human Rights Act 1998. PMID- 10821024 TI - 'Near PVS': a new medico-legal syndrome? AB - This paper addresses the current medico-legal issues surrounding PVS (Permanent Vegetative State), including: the lack of a unified definition of the acronym PVS, the varying criteria for diagnosis of PVS, and the issue of patients who maintain a minimal degree of consciousness and cannot be categorized as PVS patients. First, we analyse the differing medical definitions and criteria for diagnosis in vegetative conditions. We also ask what part 'consciousness' plays in treatment decisions made by the family, the healthcare team, and the courts, by analysing a unique Irish case of a patient in a state deemed by the courts as 'near PVS'. The paper demonstrates that there is now a legal dichotomy in vegetative patients. However, the manner in which the court treated these patients is the same. Underlying this discussion we hope to demonstrate how medical practice is subject to legal decisions and thus the importance of establishing uniform medical guidelines to assist the non-medical professional. PMID- 10821025 TI - Prediction of violence on a psychiatric intensive care unit. AB - The aim of the present study was to identify patients who were at particular risk of violent behaviour, over a period of one month in a psychiatric high dependency unit, and to examine whether particular patients could be identified at admission who would subsequently be violent. Medical records, and other documents for 52 inpatients were included. Demographic, historical and clinical data available at admission were noted. All violent incidents were recorded and classified. Seventeen of 52 patients committed physical violence, usually to the staff. Most incidents were carried out by a small number of individuals. Identifying the violent, using variables available at admission, was not possible. The ward admitted a group with a subsequent high rate of violence. Prediction within this high-risk group was not possible. Measures aimed at preventing violence on a local secure ward should apply to all patients and should not be targeted just at those that would appear to be the most likely to be violent. PMID- 10821026 TI - The informal patient in England and Wales. AB - The informal status of a patient is a very important topic because the vast majority of mental patients in hospital are informal. The origins of the status are traced to the Royal Commissions of 1924-6 and 1954-7 which recommended voluntary admissions and informal admissions, respectively. It is pointed out, inter alia, that it is only generally true to say the informal patient has consented to admission and cannot be treated without his or her consent because exceptionally he or she can be given such treatment, e.g. on the grounds of necessity, as held by the House of Lords in R v Bournewood Community and Mental Health Trust (1998) and that today there are two types of informal patients: those who can, and do, consent to admission, and those who cannot consent to admission, but do not show willingness to leave hospital. It is argued that there is only a power under the Mental Health Act 1983 to admit patients informally. Finally, the informal patient's consent to admission, consent to treatment, other rights, leaving hospital, and how his or her position can be improved are looked at. PMID- 10821027 TI - Reconviction after discharge from a regional secure unit. AB - There are at present no published data on criminal conviction following discharge from medium-secure psychiatric facilities in the United Kingdom. The results of an audit at the Kent Forensic Psychiatry Service found a 7% rate for serious offences in an average follow-up period of three years and five months, a figure similar to high-security hospital studies. Only one serious offence was unknown to the clinical team. Reports on larger samples are awaited. PMID- 10821028 TI - Detention of elderly psychiatric inpatients under section 5(2) of the Mental Health Act 1983. AB - Emergency detention of elderly psychiatric inpatients appears to have attracted very little or no attention in published studies. In this retrospective review, all applications of sections 5(2) and 5(4) of the Mental Health Act 1983, detaining elderly inpatients in North Cheshire between 1985 and 1997 were reviewed. Forty-three percent of elderly inpatients under s.5(2) regained their voluntary status, while 57% were detained under another section by the end of the 72 hours (p < 0.05). Duration in hospital prior to applying s.5(2), clinical diagnosis of functional mental illness and use of s.5(4) appear to increase the likelihood of converting s.5(2) into other sections. The high rate of non conversion of s.5(2) in the elderly to s.2 or 3 may imply that in almost half of the cases, emergency detention may have been used to control isolated incidents of disturbed behaviour in otherwise co-operative patients. Educating doctors and nurses in guidance put forward by the Code of Practice (1993) remains, probably, the main key to a better use of emergency holding powers. PMID- 10821029 TI - Aluminum phosphide fatalities, new local experience. AB - Aluminum phosphide (AlP) pesticide is a highly toxic, low cost, and easily accessible rodenticidal agent. Its toxicity results from the liberation of phosphine gas upon exposure to moisture, which leads to multisystem involvement, resulting in serious consequences. The highly toxic parathion insecticide was a common cause of mortality in pesticide fatalities, prior to its banning. Its toxicity was familiar to the public as well as to physicians. Recently, ten fatalities due to AlP were encountered within a three-month period during spring, when it was used as a rodenticide in the vicinity of grain stores. The victims' ages ranged from 1-34 years. The circumstances of death were accidental in six cases, suicidal in two and possibly homicidal in two cases. Retrospectively, the clinical manifestations, scene investigation, autopsy, histological and toxicological findings supported the diagnosis of AlP intoxication. Immediate recognition was difficult due to unfamiliarity of the agent to the physicians. The occurrence of these fatalities might suggest changes of pattern in pesticide poisoning. This should raise the attention of the physician to the problem of AlP poisoning and also necessitates the awareness of the public to the hazards of this poison. Education, proper handling, strict observation and abiding by the regulations controlling this material are good protective measures against AlP poisoning. PMID- 10821030 TI - A case of serial homicide by injection of succinylcholine. AB - The abstract of this paper was presented at the 14th Meeting of the International Association of Forensic Sciences, Tokyo in 1996. We report a bizarre criminal case of suspected serial homicide by injection of a muscle relaxant (succinylcholine). Five victims were found buried in a rural area. In two victims showing moderate decomposition (about three months after death), intense pulmonary oedema with pleural effusion was observed. Evidence of a puncture site was found in one of the victims. Succinylcholine could not be detected in the victims, but was identified in a syringe found near the corpses. The 40-mg ampule dose of succinylcholine administered intramuscularly to the victims, possibly causing prolonged apnea, was considered to be at least around the minimum lethal dose, although the combined effect of the sedation with hypnotics also used was not negligible. PMID- 10821031 TI - An autopsy case of sudden death in a patient with idiopathic scoliosis. AB - A 38-year-old woman with idiopathic scoliosis (right convex thoracic scoliosis, 78 degrees; left convex lumbar curvature, 75 degrees) died suddenly. Forensic autopsy and histopathologic examination revealed chronic congestive oedema, numerous cavities and atrophic changes of heart. These changes, including both respiratory changes and biventricular failure caused by hypoplastic cardiac changes, were due to a deformed thoracic cage. This case illustrates that not only abnormalities of respiratory function and cor pulmonare, but also hypoplastic cardiac changes, may cause sudden death in a patient with untreated scoliosis. PMID- 10821032 TI - Importance of examination of buccal cavity, trachea, hand and all injuries. AB - Four cases are discussed, with the point in view that examination of oral cavity, trachea, hands and all injuries is a vital part of post-mortem examination for the administration of justice. The cases are as follows. 1. Alleged case of death due to faulty treatment by physician, in which the death was due to choking by a medicinal tablet given forcefully to the child by his grandmother. 2. A case of hanging in which examination of the left palm revealed a ball pen testing mark on the hypothenar region. 3. In a case of homicidal death, a metal fragment was found in an incised wound which became an important piece of evidence for the administration of justice. 4. A case of hanging in which the suicide note was found in the oral cavity. PMID- 10821033 TI - [Facial dysmorphism due to complicated congenital nasolacrimal duct obstruction]. AB - Congenital nasolacrimal duct obstruction (in the literature also called congenital nasolacrimal mucocele or dacryocystocele) presents either as a simple nasolacrimal obstruction with epiphora or as a complicated obstruction on both sides of the lacrimal sac with midfacial deformation and respiratory insufficiency. Five neonates were treated for such complicated congenital nasolacrimal duct obstruction. Symptoms varied from an inner canthal purple-red swelling in the medial corner of the eye, with epiphora, to a serious midfacial deformation with respiratory insufficiency. The presence of a bilateral nasolacrimal mucocele leads to a broadened nasal bridge, which may be falsely interpreted as a dysmorphic feature of a congenital syndrome. Recognition of the condition allows a quick and efficient diagnosis and prevents unnecessary diagnostic procedures and anxiety. Differential diagnosis mainly includes frontobasal encephalocele and haemangioma. Treatment consists of massage, and catheterization if necessary. PMID- 10821035 TI - [Gastrointestinal surgery and gastroenterology. X. Obstipation: treatment]. AB - Treatment of the symptom of constipation is usually symptomatic. Correction of abnormal lifestyle and dietary habits may constitute a sufficient treatment of constipation. In addition, bulk-forming agents and osmotically active laxatives may be used. Only when these measurements are insufficiently effective, may more drastic treatment options such as contact laxatives and enemas may be used. In cases of spastic pelvic floor syndrome physiotherapy of the pelvic floor is indicated. PMID- 10821034 TI - [Gastrointestinal surgery and gastroenterology. IX. Obstipation: etiology and diagnosis]. AB - Constipation and disordered defaecation are symptoms, not disease entities. Usually these symptoms are not caused by organic abnormalities, but by disordered motility of the colon and pelvic floor. Both decreased colonic motility (inertia coli) and increased frequency and amplitude of haustrating colonic contractions in the context of an irritable bowel syndrome may lead to constipation. A third important functional cause of constipation is by paradoxical straining of the pelvic floor muscles during (attempts to) defaecate: anismus. In the diagnosis of constipation the primary aim usually is the exclusion of organic disorders. A plain abdominal X-ray and measurement of colonic transit with radiopaque particles will provide information about the severity of the constipation. Defaecography is indicated primarily if disordered faecal expulsion exists. Anorectal manometry is relevant when Hirschsprung's disease is suspected. PMID- 10821036 TI - [Methicillin-resistant Staphylococcus aureus in medical and paramedical personnel returned from work in a foreign hospital]. AB - The cultures of two patients of the Surgical Intensive Care Unit (IC) of the Medical Centre of Utrecht University were found positive for methicillin resistant Staphylococcus aureus (MRSA). A male nurse turned out to be the source, 4 months after his return from working in an English hospital. Cultures were, by mistake, not taken directly on arrival from abroad. Pulsed-field gel electrophoresis proved MRSA strains from both source and the 2 patients to be identical to a strain which was epidemic in Great Britain but had never occurred in the Netherlands. The IC has meanwhile been closed; at source investigation, 14 other patients and six staff members were found MRSA-positive. The policy in the hospital is to screen health care workers for MRSA carriership on return from an hospital abroad. The success of the policy depends strongly on the cooperation of health care workers in this matter. PMID- 10821037 TI - [Terminologia anatomica: a new reference work for anatomy]. AB - The Nomina anatomica (6th edition in 1989) was the standard anatomical nomenclature until recently. It has been succeeded by the Terminologia anatomica (TA) by resolution of the International Federation of Associations of Anatomists in 1999. By now the TA has been published as a book and on cd-rom. The objective of the TA is to further uniformize the anatomical terminology, also internationally, and especially to democratize it, i.e. to ensure that it accommodates the needs of clinicians and medical scientists. The number of anatomical terms listed has grown to more than 7500. The increase is caused mainly by the insertion of clinical anatomical and neuro-anatomical names. A number of incorrect or misleading terms have been replaced. Each Latin term has a unique code number and is supplied with an English equivalent (International anatomical terminology; IAT) the spelling of which in either UK English or American English is considered equally correct. Based on the TA every language can formulate its own anatomical nomenclature. The nomenclature is presented per system or organ tract. An alphabetic index follows TA as well as the English IAT list. The use of eponyms is discouraged, but a list of well-known ones is appended to facilitate accessibility of older literature. Relevant suggestions about amendments are eagerly awaited and a broad basis of endorsement among the medical scientific world is hoped for. This journal will adopt the TA/IAT from now on as the work of reference regarding anatomical terms. PMID- 10821038 TI - [The influence of urinary incontinence on quality of life of community-dwelling, 45-70 year old Dutch women]. AB - OBJECTIVE: To establish the prevalence of urinary incontinence in non institutionalized Dutch women and determine the consequences of urinary incontinence for their quality of life. DESIGN: Cross sectional questionnaire survey. METHOD: A random sample of 1905 women aged 45-70 years of the population of Zeist, the Netherlands, were sent a questionnaire. Generic quality of life was measured with the RAND-36 and disease specific quality of life was measured with a Dutch translation of the Urogenital Distress Inventory (UDI) and Incontinence Impact Questionnaire (IIQ). Based on the answers to the questionnaire four groups of women were formed: no incontinence, only stress, only urge or a combination of stress and urge incontinence. These 4 groups were compared as to questionnaire scores. RESULTS: A total of 1086 questionnaires could be analysed. The prevalence of urinary incontinence was 57.1%: 28.7% stress incontinence, 5.6% urge incontinence and 22.7% both. Of those who reported urinary incontinence 6% were severely inconvenienced by it. Women with urinary incontinence reported a decrease in physical functioning and vitality as compared with women without incontinence (RAND-36). Especially women with urge or a combination of stress and urge incontinence had more severe impairment of their quality of life as compared to women with only stress incontinence. CONCLUSION: The prevalence of urinary incontinence is higher than it is usually reported. Especially the urge component affects the quality of life in a negative way. PMID- 10821039 TI - [Link between indication for cesarean section and need for resuscitation of the neonate]. AB - OBJECTIVE: To study the relationship between the indication for caesarean section (CS) and the need for resuscitation of the infant. DESIGN: Prospective cohort study. METHODS: During one year the following information was collected for each CS in a secondary teaching hospital (Isala klinieken/Weezenlanden Hospital), Zwolle, the Netherlands: indication for the CS, Apgar scores after 1 and 5 min and measures taken by the attending paediatrician at birth. These measures were divided into two groups: resuscitation (bag and mask ventilation or endotracheal intubation and ventilation) or no resuscitation (stimulation, supplemental oxygen, or no action at all). RESULTS: During the study year 202 CS were carried out. In the 50 CS carried out because of a narrow pelvis or a previous CS and in the 61 CS carried out for non-progressive labour there was little need for resuscitation of the newborn (4 and 3.3%, respectively), similar to resuscitation requirements after vaginal birth. The need for resuscitation of the newborn was significantly greater in the other indication groups, namely in 6/37 (16%) of cases of foetal malposition, in 3/10 (30%) of placental dysfunction, and in 11/44 (25%) of foetal distress. One infant had to be intubated directly after birth. The 1-minute Apgar score was lower in infants born after CS under general anaesthesia than under spinal anaesthesia (p = 0.002), regardless of the indication for the CS. CONCLUSION: Based on the indication for a CS, a paediatrician or physician experienced in neonatal resuscitation is required for a high-risk CS (foetal malposition, placental dysfunction, foetal distress, and general anaesthesia). PMID- 10821041 TI - [Objective evaluation procedures in cases of possible diagnostic error]. PMID- 10821040 TI - [Hypocalcemia as a cause of reversible heart failure]. AB - A 72-year-old woman with therapy resistant congestive heart failure presented with severe hypocalcaemia due to hypoparathyroidism after strumectomy more than 25 years before. After suppletion of calcium her complaints resolved and there was considerable improvement in left ventricular function. Our case report suggests that hypocalcaemia induced cardiomyopathy should be considered in the differential diagnosis of therapy resistant heart failure and that myocardial impairment is reversible after administration of calcium. PMID- 10821042 TI - Effect of sesquiterpene lactones on antioxidant enzymes and some drug metabolizing enzymes in rat liver and kidney. AB - Previously we have reported that several sesquiterpene lactones isolated from Helenium aromaticum and Telekia speciosa showed pro-oxidative properties and caused glutathione level depletion in rat liver in vivo. In the present study we examined the in vivo effect of these lactones on antioxidant enzyme systems and some drug metabolizing enzymes in the liver and the kidney of rats. We found that the majority of the compounds increased the hepatic activity of glutathione peroxidase (GPx), glutathione reductase (GR), and catalase (CAT), but superoxide dismutase (SOD) activity was distinctly lowered by five lactones. A few of the compounds tested caused a decrease in the hepatic cytochrome P450 content and reduced the activity of NADPH-cytochrome P450 reductase, aminopyrine demethylase, aniline hydroxylase and glutathione-S-transferase. Results for the kidney showed fewer changes in activities of both classes of enzymes when compared to the liver. Not all lactones affected the enzymes under test, the most active were: linifolin, helenalin, mexicanin 1 and telekin. 6 alpha-Hydroxy-2,3 dihydroaromaticin behaved differently towards monooxygenases since it induced the activity of aminopyrine demethylase and aniline hydroxylase. PMID- 10821043 TI - Inhibition of serine proteases by anti-inflammatory triterpenoids. AB - The lupane triterpenoid lupeol, the ursane triterpenoid alpha-amyrin and esters of these compounds are present in the bark of roots of Alstonia boonei (Apocynaceae) and have anti-inflammatory properties. alpha-Amyrin is a competitive inhibitor of bovine trypsin and chymotrypsin (Ki values 29 microM and 18 microM, respectively). Lupeol linoleate, lupeol palmitate and alpha-amyrin linoleate are non-competitive inhibitors of trypsin (Ki values 7 microM, 10 microM and 16 microM, respectively). alpha-Amyrin linoleate is also a non competitive inhibitor of chymotrypsin (Ki value 28 microM). Lupeol is a competitive inhibitor of both trypsin and chymotrypsin (Ki values 22 and 8 microM, respectively). alpha-Amyrin palmitate is a potent non-competitive inhibitor of chymotrypsin (Ki 6 microM). Lupeol, alpha-amyrin and the palmitic and linoleic acid esters of these compounds are ineffective or very weak as inhibitors of porcine pancreatic elastase and of Lucilia cuprina and Helicoverpa punctigera leucine aminopeptidases. These hydrophobic triterpenoids represent further examples of anti-inflammatory triterpenoids that are PKA inhibitors as well as being selective protease inhibitors. PMID- 10821044 TI - Yangambin, a lignan obtained from Ocotea duckei, differentiates putative PAF receptor subtypes in the gastrointestinal tract of rats. AB - We investigated the presence of PAF receptor subtypes in the tissues of the gastrointestinal tract, airways, blood vessels and in murine macrophages. For this purpose we have used a competitive PAF receptor antagonist, yangambin (YAN), extracted from the Brazilian plant "louro de cheiro" (Ocotea duckei Vattimo). Rat duodenum, jejunum, ileum, colon, stomach fundus, trachea and bronchia were removed and 1.5-2 cm muscle segments from those regions were mounted in a 10 ml organ bath with aerated physiological solution at 37 degrees C. PAF evoked a contraction of the rat jejunum, ileum, colon and stomach fundus. The contraction was slow and resistant to wash and was followed by desensitization to further doses of PAF. Contractions induced by PAF (10(-6) M) were inhibited by YAN (10( 7) to M-2 x 10(-5) M) and WEB 2086 (10(-6) m to M-5 M) in rat jejunum, ileum and colon but not in the stomach fundus. In the rat stomach fundus only WEB 2086 (5 x 10(-6) M) was able to block PAF-induced contraction. The contractions induced by acetylcholine, histamine, 5-hydroxytryptamine and vasopressin were not inhibited by prior administration of YAN. Yangambin also significantly inhibited PAF induced vascular permeability in rat duodenum, jejunum, ileum, colon, and mesentery. Yangambin significantly inhibited PAF-induced lipid body formation in mice peritoneal macrophages. We suggest that YAN is a selective PAF antagonist which is able to discriminate putative PAF receptors subtypes present in the stomach fundus. PMID- 10821045 TI - Inhibition of Helicobacter pylori hemagglutination by polysaccharide fractions from roots of Panax ginseng. AB - Polysaccharides from the roots of Panax ginseng were extracted by hot water and fractionated by using ethanol precipitation and ion exchange chromatography. Fractions FC (crude extract), F1 (fraction precipitated by ethanol), F1N (fraction unbound to DEAE-Sepharose CL-6B), and F1A (bound fraction) were obtained. Their carbohydrate analyses showed that acidic fraction F1A contains higher amounts of galactose, arabinose and uronic acids, in comparison to FC and F1. Fraction F1N mainly consists of glucose. The inhibition of Helicobacter pylori-induced hemagglutination revealed different inhibitory activities of these fractions. In particular, acidic fraction F1A showed a remarkable inhibitory activity (minimum inhibition concentration was 0.25 mg/ml) among the polysacharide fractions. However, digestion of the fraction F1A with pectinase resulted in a lower molecular weight oligosaccharide fraction F1AP which was non inhibitory at the concentration of 4 mg/ml. Comparison of inhibitory activities and carbohydrate compositions of isolated fractions indicates that the activity correlated with the contents of galactose, arabinose, and uronic acids. These data suggest that acidic polysaccharides may be responsible for the inhibitory activity. PMID- 10821046 TI - In vivo studies on the anti-inflammatory activity of pachymic and dehydrotumulosic acids. AB - Pachymic and dehydrotumulosic acids were studied in different models of acute and chronic inflammation. They proved to be active in most of the methods applied. None of them were active against arachidonic acid-induced ear edema. Dehydrotumulosic acid significantly diminished the mouse ear edema induced by ethyl phenylpropiolate, while pachymic acid was ineffective. When the putative corticoid-like mechanism of both compounds was explored, pachymic acid activity was partially abolished by the glucocorticoid receptor antagonist progesterone, but dehydrotumulosic acid activity was not affected. In vivo experiments demonstrated the inhibition by both principles of the phospholipase A2 (PLA2) induced extravasation. The previous report on the effects of both compounds in vitro against PLA2, together with the present in vivo results, support the idea that the inhibition of this enzyme probably constitutes their main mechanism of action. PMID- 10821047 TI - Caffeic acid as active principle from the fruit of Xanthium strumarium to lower plasma glucose in diabetic rats. AB - The antihyperglycemic effect of caffeic acid, one of the phenolic compounds contained in the fruit of Xanthium strumarium, was investigated. After an intravenous injection of caffeic acid into diabetic rats of both streptozotocin induced and insulin-resistant models, a dose-dependent decrease of plasma glucose was observed. However, a similar effect was not produced in normal rats. An insulin-independent action of caffeic acid can thus be considered. Otherwise, this compound reduced the elevation of plasma glucose level in insulin-resistant rats receiving a glucose challenge test. Also, glucose uptake into the isolated adipocytes was raised by caffeic acid in a concentration-dependent manner. Increase of glucose utilization by caffeic acid seems to be responsible for the lowering of plasma glucose. PMID- 10821048 TI - Reproductive toxicity of piperine in Swiss albino mice. AB - Piperine (CAS 94-62-2) is a constituent of various spices which are used as common food additives all over the world. The reproductive toxicity of piperine was studied in Swiss albino mice. Relevant short-term tests were employed to assess the effect on estrous cycle, mating behaviour, toxicity to male germ cells, fertilization, implantation and growth of pups. Piperine (10 and 20 mg/kg b.w.) increased the period of the diestrous phase which seemed to result in decreased mating performance and fertility. Post-partum litter growth was not affected by the piperine treatment. Sperm shape abnormalities were not induced by piperine at doses up to 75 mg/kg b.w. Considerable anti-implantation activity was recorded after five days post-mating oral treatment with piperine. The sex ratio and post-implantation loss were unaffected after treatment with piperine. Intrauterine injection of piperine caused the total absence of implants in either of the uterine horns (16.66%) or one of the horns (33%) of treated females. No histopathological changes were detected in the ovary and the uterus at the cellular level. Prostaglandin E1-induced acute inflammation of rat paw was significantly reduced after piperine treatment. Our results show that piperine interferes with several crucial reproductive events in a mammalian model. PMID- 10821049 TI - Haploid plants regenerated from androgenic cell cultures of Digitalis lanata. AB - Androgenic callus was obtained from cold treated anthers and pollen of Digitalis lanata. The callus was mixoploid and contained haploid, diploid and tetraploid cells as shown by impulse cytophotometry. Haploid cell lines were selected by colony cloning. They were unstable and selection had to be repeated every 1-2 months. Mixoploid shoot cultures were derived from embryogenic haploid cell lines via somatic embryos. Haploid shoots were selected by explanting shoot tips. The shoots showed wide variability in cardenolide content and profile. Rooting of the haploid shoots resulted in haploid plants. These plants were smaller in size than diploid plants. Often the flowers were morphologically abnormal and showed male sterility due to crippled anthers. PMID- 10821050 TI - Light-mediated antifungal activity of Echinacea extracts. AB - This study demonstrated that plant extracts containing acetylenic isobutylamides and polyacetylenes, previously reported as occurring in Echinacea, have phototoxic antimicrobial activity against fungi, including clinically relevant pathogenic fungi. Results show that hexane extracts of Echinacea variably inhibit growth of yeast strains of Saccharomyces cerevisiae, Candida shehata, C. kefyr, C. albicans, C. steatulytica and C. tropicalis under near UV irradiation (phototoxicity) and to a lower extent without irradiation (conventional antifungal activity). The presence of polyacetylenes and alkylamides in extracts of different organs was confirmed in Echinacea purpurea by HPLC in agreement with previously reported data in the literature, and was related to phototoxic activity. Two representative pure compounds, undeca-2E,4Z-diene-8,10-diynoic acid isobutylamide and dodeca-2E,4E,8Z,10E/Z-tetraenoic acid isobutylamide, were isolated from Echinacea purpurea root extracts, and compared in a disk assay (5 micrograms/disk) with the highly conjugated trideca-1-ene-3,5,7,9,10-pentayne (previously isolated in our laboratory and found here in E. purpurea). Significant phototoxicity was demonstrated by pure trideca-1-ene-3,5,7,9,10 pentayne, while only minor phototoxicity was induced by the other two acetylenic compounds. Phototoxic activity of Echinacea spp. is primarily attributed to the ketoalkenes and ketoalkynes abundantly present in the roots. PMID- 10821051 TI - Parameters influencing the yield and composition of the essential oil from Cretan Vitex agnus-castus fruits. AB - Mature and immature fruits of a Cretan Vitex agnus-castus L. population were chosen to investigate different parameters such as comminution, maturity, distillation period and extraction method influencing the essential oil yield and composition. The effect of the comminution and the maturity of the plant material showed highly significant differences in yield and composition of the essential oils obtained, as well as the distillation duration from one to five hours and the method applied (hydrodistillation and simultaneous distillation extraction). The variation of 36 essential oil components due to the parameters applied was studied. The results showed that many different essential oil qualities can be obtained from the same plant material according to the parameters employed in its extraction. Entire fruits hydrodistilled for one hour yielded an oil much richer in monoterpene hydrocarbons and oxygenated compounds whereas the best combination to obtain an oil rich in less volatile compounds is by SDE of comminuted fruits for five hours. For mature fruits the main components varied as follows due to the parameters studied: sabinene 16.4-44.1%, 1,8-cineole 8.4-15.2%, beta caryophyllene 2.1-5.0%, and trans-beta-farnesene 5.0-11.7%. PMID- 10821052 TI - Anti-platelet aggregation alkaloids and lignans from Hernandia nymphaeifolia. AB - A new aporphine, N-(N-methylcarbamoyl)-O-methyl-bulbocapnine (1), together with seven known compounds, (-)-5'-methoxypodorhizol (2), a mixture of beta sitosterone (3) and stigmasta-4,22-dien-3-one (4), a mixture of 3 beta hydroxystigmast-5-en-7-one (5) and 3 beta-hydroxystigmasta-5,22-dien-7-one (6), and a mixture of 6 alpha-hydroxystigmast-4-en-3-one (7) and 6 alpha hydroxystigmasta-4,22-dien-3-one (8), were isolated in continuing studies on the trunk bark of Formosan Hernandia nymphaeifolia. The structures of these compounds were determined through spectral analyses. In addition, the previously reported six alkaloids, laurotetanine, oxohernagine, thalicarpine, reticuline, (+) vateamine-2'-beta-N-oxide, (+)-hernandaline and six lignans, (+)-epiaschantin, (+)-epimagnolin, (+)-epiyangambin, (-)-hernone, (-)-yatein, (-) deoxypodophyllotoxin were demonstrated to have anti-platelet aggregation activity. PMID- 10821053 TI - New phenylethanoids from Buddleja cordata subsp. cordata. AB - Bioassay-guided fractionation of a crude extract of the stem bark of Buddleja cordata subsp. cordata with significant antimycobacterial activity led to the isolation of a mixture composed by ten new long-chain esters of 2[4' hydroxyphenyl]-ethanol (1-10), along with the lichen metabolites methyl beta orcinolcarboxylate (11) and beta-orcinolcarboxylate (12). Extensive HPLC allowed the separation of the major components of the mixture, which were characterized by spectral means as 2[4'-hydroxyphenyl]-ethyl stearate (3), 2[4'-hydroxyphenyl] ethyl behenate (6), and 2[4'-hydroxyphenyl]-ethyl lignocerate (8). The minor esters were identified as 2[4'-hydroxyphenyl]-ethyl palmitate (1), 2[4' hydroxyphenyl]-ethyl heptadecanoate (2), 2[4'-hydroxyphenyl]-ethyl nonadecanoate (4), 2[4'-hydroxyphenyl]-ethyl arachidate (5), 2[4'-hydroxyphenyl]-ethyl tricosanoate (7), 2[4'-hydroxyphenyl]-ethyl pentacosanoate (9), and 2[4' hydroxyphenyl]-ethyl hexacosanoate (10) by GC-MS analysis of the methyl esters derivatives of the fatty acids obtained by alkaline hydrolysis of the mixture. Compound 8 exhibited moderate antibacterial activity against Mycobacterium tuberculosis (MIC = 64 micrograms/ml). PMID- 10821054 TI - New antimalarial and cytotoxic sungucine derivatives from Strychnos icaja roots. AB - Reinvestigation of Strychnos icaja Baillon resulted in the isolation of vomicine, isostrychnine and of three new sungucine derivatives, named isosungucine (8), 18 hydroxy-sungucine (9) and 18-hydroxy-isosungucine (10). They were identified by detailed spectroscopic methods. The complete 1H- and 13C-NMR study of sungucine was also realized. Some of these compounds were highly active against Plasmodium falciparum in vitro and more particularly against the chloroquine-resistant strain. Compound 10 showed a selective antiplasmodial activity, with > 100-fold greater toxicity towards Plasmodium falciparum, relative to cultured human cancer cells (KB and HeLa lines) or fibroblasts (WI38). PMID- 10821055 TI - Discrimination among three species of medicinal Scutellaria plants using RAPD markers. AB - An analysis of random amplified polymorphic DNA (RAPD) was performed using nine accessions of three species of medicinal plants in the genus Scutellaria (S. galericulata, S. lateriflora and S. baicalensis; known collectively as skullcap) in an effort to distinguish between members of these three species. Dried aerial parts of the two species S. galericulata and S. lateriflora are difficult to distinguish morphologically. Ten arbitrary primers produced 92 fragments, and eight of the primers yielded 23 species-specific fragments among the three species. Six fragments were specific for S. galericulata, seven for S. lateriflora and ten for S. baicalensis. When primers A02 and A06 were used in the polymerase chain reaction, RAPD fragments that were specific for each of the three species were generated simultaneously. Primer A02 produced five species specific fragments: one was specific for S. galericulata; two for S. lateriflora; and two for S. baicalensis. Primer A06 produced three species-specific fragments: one for S. galericulata; one for S. lateriflora; and one for S. baicalensis. The RAPD markers that were generated with these two primers should rapidly identify members of the three species of Scutellaria. The consistency of the identifications made with these species-specific RAPD markers was demonstrated by the observation that each respective marker was generated from three accessions of each species, all with different origins. Furthermore, cluster analysis using the 92 RAPD fragments produced a dendrogram of genetic relatedness that was in good agreement with the taxonomic designations of the three species. Thus, the RAPD markers should be useful for the future identification of members of the three species of medicinal Scutellaria plants. PMID- 10821056 TI - alpha-Glycosidase inhibitory activity of hexagalloylglucose from the galls of Quercus infectoria. AB - Hexagalloylglucose (3-O-digalloyl-1,2,4,6-tetra-O-galloyl-beta-D- glucose), which was isolated from the methanol extract of the galls of Quercus infectoria, significantly inhibited alpha-glycosidases such as sucrase, maltase and isomaltase. Its inhibitory activity was comparable to acarbose being used as a hypoglycemic agent, while the inhibitory activity on alpha-amylase was approximately 10 times lower than that of acarbose. The results indicate that, when compared to acarbose, hexagalloylglucose might reduce the side effects by reducing inhibition of alpha-amylase. PMID- 10821057 TI - Tyrosinase inhibitors from Artocarpus gomezianus. AB - Eight compounds including, phenyl-beta-naphthylamine (1), isocyclomorusin (2), cycloartocarpin (3), artocarpin (4), norartocarpetin (5), cudraflavone C (6), albanin A (7), and resveratrol (8) were isolated from the roots of Artocarpus gomezianus. Compounds 5 and 8 exhibited potent tyrosinase inhibitory activity. The 1H- and 13C-NMR properties of 1, 3 and 8 were extensively studied. PMID- 10821058 TI - Coumarins and carbazoles with antiplasmodial activity from Clausena harmandiana. AB - Activity guided fractionation of extracts from Clausena harmandiana have led to the identification of four known compounds, heptaphylline (1), clausine K (2), dentatin (5), and clausarin (6). All these compounds, except clausine K (2), exhibited antiplasmodial activity against Plasmodium falciparum. While the new dimethylated derivative 4, derived from 2, showed no antiplasmodial activity, the monomethylated product 3 (clausine H) exhibited activity comparable to that observed for compounds 1 and 5. PMID- 10821059 TI - Anti-inflammatory activity of coumarins from Decatropis bicolor on TPA ear mice model. AB - From the aerial parts of Decatropis bicolor, heraclenin (1), seselin (2), psoralen (3), imperatorin (4), skimmianine (5), and heraclenol (6), were isolated. This is the first time that coumarin-like compounds are isolated from Decatropis genus. The anti-inflammatory properties of compounds 1-6 were examined against the ear edema in mice produced by TPA. The results suggest that the anti inflammatory activity of each compound depends of its individual substitution on the aromatic ring rather than the coumarin skeleton itself. PMID- 10821060 TI - Aquaticol, a bis-sesquiterpene and iridoid glucosides from Veronica anagallis aquatica. AB - Aquaticol, an unusual bis-sesquiterpene was isolated from the medicinal plant Veronica anagallis-aquatica, in addition to 11 known compounds, aucubin, geniposidic acid, mussaenoside, catalposide, verproside, amphicoside, catalpol, boschnaloside, shanzhiside methyl ester, sitosterol and beta-stigmast-4-en-6 beta ol-3-one. The structure of aquaticol was determined by spectroscopic means and by X-ray crystallographic analysis. PMID- 10821061 TI - Anti-inflammatory and analgesic activities of the latex containing triterpenes from Himatanthus sucuuba. AB - Some triterpenes and iridoids were previously isolated from the stem bark of Himatanthus sucuuba. The latex from Himatanthus sucuuba is used in popular amazonian medicine as an anti-inflammatory remedy. Fractions of the latex were pharmacologically evaluated with a view to verify this popular use in the carrageenan-induced rat paw edema and in the acetic acid-induced mouse constriction tests. The hexane fraction inhibited the edema formation by 35.9% at a dose of 200 mg/kg (p.o.) but no activity was observed at 100 mg/kg (p.o.). The triterpenes present in the hexane fraction were identified as lupeol acetate, alpha-amyrin and lupeol cinnamates. The fraction containing only cinnamates inhibited the edema and the abdominal constrictions by 50-40% and 57.9%, respectively, at 100 mg/kg (p.o.). Among all the fractions studied, the fraction containing only cinnamates showed the greatest anti-inflammatory activity which suggests that these compounds were responsible for the previously described activity of the crude extract. PMID- 10821062 TI - Antiplatelet and antithrombotic effects of the diterpene spiramine Q from Spiraea japonica var. incisa. AB - Spiramine Q, a diterpene, was isolated from a Chinese herbal plant Spiraea japonica var. incisa Yu. Born's and Wan HY's methods were used to investigate effects of spiramine Q on rabbit platelet aggregation and serotonin release, respectively. Its antithrombotic effect in mice was also evaluated by Myers' method. Spiramine Q selectively inhibited arachidonic acid-induced platelet aggregation in vitro or ex vivo, and decreased serotonin secretion from rabbit platelets. Spiramine Q (5 mg/kg) decreased the mouse mortality caused by injection of 80 mg/kg arachidonic acid in the tail vein. The results suggested that spiramine Q showed potent antiplatelet and antithrombotic activites. PMID- 10821063 TI - Immunomodulating properties of the diterpene andalusol. AB - Andalusol [ent-13(16),14-labdadiene-6 alpha,8 alpha,18-triol], a labdane diterpene isolated from an anti-inflammatory extract of the medicinal plant Sideritis foetens Clem. was evaluated for immunomodulating properties. It exhibited a dose-dependent inhibitory effect on the hemolytic activity of the classical complement pathway and also inhibited lymphocyte proliferation induced by concanavalin A at non-cytotoxic concentrations (1-100 microM). No effect on the alternative pathway was observed. These results indicate that andalusol has immunosuppressive effects in vitro in addition to its anti-inflammatory activity. PMID- 10821064 TI - Diterpenoids from Euphorbia peplus. AB - From a pro-inflammatory active extract of Euphorbia peplus, two new diterpene polyesters based on the pepluane and jatrophane skeletons were isolated, together with four known ingenane and jatrophane diterpenes. The structures were determined on the basis of extensive NMR studies. Ingenol 3-angelate, which was obtained for the first time from this plant, is an irritant toxin with high activity. PMID- 10821065 TI - Three new taxoids from the seeds of Japanese yew, Taxus cuspidata. AB - Five taxane diterpenoids were isolated from the seeds of the Japanese yew, Taxus cuspidata Sieb. et Zucc. The structures were established as 2 alpha,7 beta,13 alpha-triacetoxy-5 alpha-(3'-dimethylamino-3'-phenyl)-propionyloxy-2(3-->20) aboe+ ++-taxa-9,10-dione (1) and 2 alpha,7 beta,9 alpha,10 beta-tetraacetoxy-5 alpha-[(2R,3S),N,N-dimethyl-3-phenylisoseryloxy]-taxa-4(20),11- dien-13-one (2), 2 alpha, 9 alpha, 10 beta-triacetoxy-5 alpha-cinnamoxy-taxa-4(20),11-diene-13 alpha-ol (3), taxezopidine J (4), and taxuspine D (5) on the basis of 1D, 2D NMR, and MS spectral analysis. Compounds 1, 2, and 3 are new compounds. PMID- 10821067 TI - The appearance of evil. PMID- 10821066 TI - Comprehensive spectroscopic investigation of alpha-onocerin. AB - A detailed NMR and MS analysis of alpha-onocerin was performed in order to provide precise phytochemical reference for structure identification and quality control of this analytical lead compound from Ononis spinosa the roots of which are widely used as a diuretic drug. Unambiguous 1H- and 13C-NMR assignments revealed that the linkage of two subunits were confirmed to possess identical stereochemistries in this triterpenoid. PMID- 10821068 TI - Health care is online to stay. PMID- 10821069 TI - Health care on a tightrope: is there a safety net? Part I: Uncompensated care. PMID- 10821070 TI - Loss prevention case of the month. History--examination--suspicion. PMID- 10821071 TI - Nontreatment variables affecting return-to-work in Tennessee-based employees with complaints of low back pain. AB - Disability and health care-related costs continue to rise as a result of work related low back injury. Our investigation examined treatment-independent variables that influenced return-to-work outcome in a sample of workers employed in Northeast Tennessee. METHODS: The review collected 11 variables from two different outpatient physical therapy clinics utilizing a balanced quota sampling design. The patients were enrolled if the documented complaint was low back pain and was an employment-related injury. The patients were grouped according to whether or not they returned to full-time pre-injury work. Twenty-five patients were enrolled in the positive outcome group, those who returned to full-time pre injury work. Twenty-two patients who did not achieve this goal were enrolled in a separate group. RESULTS: Return-to-work for these patients was not dependent upon age, gender, insurer, number of physical therapy treatments attended, or previously reported low back injury. Those who returned to work had (1) a higher percentage of patients working full-time at their pre-injury position during the rehabilitation process (28% vs. 0%); (2) a higher compliance with the treatment schedule (97% vs. 93%); (3) a lower cancellation rate (0.5 vs. 2.4); (4) a shorter interval in days between reporting the injury and initiation of physical therapy rehabilitation (27 vs. 58); and (5) a lower percentage of previous surgeries resulting from low back injuries (12% vs. 36%), than those who did not. A relationship was also demonstrated between previous surgery and the interval prior to beginning treatments (P < or = 0.0001). However, no relationship was observed between previous surgery and compliance, or between the interval prior to beginning treatments and compliance. DISCUSSION: These results document two variables representing independent factors affecting return-to-work in this population. The first was previous injury influencing the current injury, as documented by both previous surgery and the interval between the current injury and beginning of treatments. The second was compliance with the treatment schedule for the current injury. The psychosocioeconomic aspects of these results are discussed. PMID- 10821072 TI - Bilateral adrenal cortical hyperplasia. PMID- 10821073 TI - Tennessee's medical examiner system. PMID- 10821074 TI - Tularemia, Kosovo. PMID- 10821075 TI - Progress towards global poliomyelitis eradication, 1999. PMID- 10821076 TI - Dracunculiasis. PMID- 10821077 TI - A comparative study of perception of sickle cell anaemia by married Nigeria rural and urban women. AB - Environmental factors may influence perception of or attitude to chronic disorders. The perception of sickle cell anaemia (SCA by 165 married Nigerian rural and 507 urban women was studied to determine how living in an urban or rural environment may influence perception. None of the subjects had children with SCA. The instrument used for data collection was a structured questionnaire designed to enquire into their knowledge about the cause, precipitating factors for crises, clinical features of SCA and their opinions regarding traditional and modern treatment options for the disorder. As a group, urban women had better knowledge about SCA than rural women probably because their social environment afforded a wider scope for interaction with and information exchange among people. For most respondents, the educational institutions attended the health institutions in the locality and the electronic media were poor sources of information on SCA. The study showed a serious lack of information about important aspects of SCA among rural women. We think the training of primary health care providers as counsellors on SCA, the inclusion of instruction about SCA in the curriculum of schools and sustained outreach programmes on SCA on the electronic media would ensure early education of people in both rural and urban communities and help to improve perception of the disorder. PMID- 10821078 TI - Retinoblastomas in Ibadan Nigeria: II--Clinicopathologic features. AB - Retinoblastomas are primary malignant intraocular neoplasms of childhood displaying photoreceptor differentiation. The present study reviews clinicopathological features of these neoplasms in Ibadan, Nigeria. Clinical, surgical pathology, and histological material of 44 histologically verified cases of retinoblastoma indexed in the Cancer Registry, University of Ibadan were analyzed. Retinoblastomas accounted for 1.1% of all malignant neoplasms and were equally frequent in male and female children. The mean age of our patients at the time of diagnosis was 32.3 months, which exceeds a corresponding age of 16-21.5 months recorded among Caucasian children. Leukocoria was the most common clinical manifestation. Eighteen percent of our patients had bilateral neoplasms. These patients were significantly younger than those with unilateral retinoblastomas. Flexner-Wintersteiner rosettes occurred in 61% and optic nerve involvement in 78% of the cases and the sites of predilection were regional lymph nodes, scalp, jaw, brain, skull, long bones, and gum. PMID- 10821079 TI - A comparative clinical evaluation of econazole nitrate, miconazole, and nystatin in the treatment of vaginal candidiasis. AB - A controlled triple open clinical study on three vaginal tablets--Econazole nitrate, Miconazole and Nystatin, was carried out. Seventy-five patients, aged between 18 and 45 years, presenting with mycologically proven cases of vaginal candidiasis, met certain set criteria and were admitted into the study. Twenty five patients were randomly assigned to each of the three treatment groups. Results of the study showed that at the 4th week after treatment, Econazole was comparable in antifungal action to Miconazole (x2 = 0.2128; p > 0.05) but significantly more antifungal than Nystatin (x2 = 8.8540; P < 0.05), although the overall clinicomycological assessment of the drugs showed no significant difference in their ratings (F = 21.34; P > 0.05). PMID- 10821080 TI - Correlation of plain radiological diagnostic features with antral lavage results in chronic maxillary sinusitis. AB - To justify the continued relevance of plain radiography of maxillary sinus in establishing diagnosis and planning treatment of chronic maxillary sinusitis, its specificity and predictive values must have a high degree of confidence. The objective of this study was to examine different plain radiographic features and correlate them with antral lavage results in patients suffering from chronic maxillary sinusitis. Air-fluid level and antral opacity have specificity of 92.3% each and positive predictive value of 87.5% and 96.0% respectively. Gross mucosal thickening and haziness showed low specificity and predictive values of 36.7% and 29.4% respectively. Normality of radiographs was reliable in predicting clear antral lavage effluent. Plain radiograph of the maxillary sinus is still relevant in establishing diagnosis and planning treatment in chronic maxillary sinusitis. PMID- 10821081 TI - Misdiagnosis of stroke--a computerised tomography scan study. AB - Misdiagnosis of stroke has significant implication for definitive therapy. This study assessed the frequency of misdiagnosis of stroke using computerised tomography (CT) scan of the brain. One hundred and fifty-six patients admitted over a five year period (1991-1996) with clinical features suggestive of stroke had their CT brain scan reviewed. Only 89 (57%) had neuroradiological features consistent with stroke, of which 59 (66%) had cerebral infarction while 30 (34%) had cerebral haemorrhage. In 67 (43%) of the cases, there were no features of cerebro-vascular accident (CVA) on the CT scan. In this group, cerebral atrophy was the commonest radiological abnormality (21/67), followed by brain tumor (10/67) and subdural haematoma (9/67). The CT scan was normal in 25 patients. The misdiagnosis of surgically treatable conditions in as many as 21 (13.5%) of the 156 patients calls for better neurological evaluation of patients admitted for stroke in tertiary and non-tertiary hospitals. Where available, patients with clinical diagnosis of stroke should have CT scan evaluation to ensure that patients who can be helped surgically are identified early and appropriately treated. PMID- 10821082 TI - Prevalence of rubella-IgG antibody in women of childbearing age in Lagos, Nigeria. AB - The prevalence of Rubella-IgG antibody was assessed in 152 Nigerian women consisting of 62 pregnant women. 82 non-pregnant women of childbearing age and 7 women who had suffered miscarriage. Rubella-IgG antibody was detected in sera of the female subjects by Immunocomb II Rubella IgG test kit (Orgenics, Israel) which is an indirect solid face Immunoassay (EIA). Rubella-IgG antibody was detected in 76% of pregnant women, 77% of non-pregnant women of childbearing age and 86% of women who had miscarriage. Similarly, the Rubella-IgG antibody titres of between 15 iu/ml and 120 iu/ml was encountered among the female subjects. Thus in general 77% of all the female subjects were positive for Rubella-IgG antibody while 23% had no detectable Rubella IgG antibody. There is need to protect this set of people considering the potential dangers of congenital anomalies associated with Rubella infection during pregnancy. The implications of the presence or absence of Rubella IgG antibody and titres are discussed. PMID- 10821083 TI - The effect of glycaemic control on the prevalence and pattern of dyslipidaemia in Nigerian patients with newly diagnosed non insulin dependent diabetes mellitus. AB - Dyslipidaemia (DL) is a common condition in patients with NIDDM, but its prevalence and the effect of glycaemic control on the disorder have only been scantily reported in Nigerians. The present study is therefore aimed at determining the effect of diabetic control on prevalence and pattern of DL in Nigerian patients with NIDDM. Thirty six diabetics were followed up for 24 weeks. Indices determined included anthropometric measurements, fasting (FBG) and two hour post prandial blood glucose (2 hours PPBG), together with glycated haemoglobin (GHb) levels, and fasting lipids at presentation, 12 and after 24 weeks of treatment. The prevalence rates of raised total cholesterol/high density lipoprotein cholesterol (TC/HDL) ratio reduced HDL-cholesterol and mixed DL decreased significantly between 0-week and 24 weeks of treatment (57.1% vs 14.3% 50% vs 11.4% and 44% vs 22.2% respectively, P < 0.001 for each). The proportion of patient with elevated low-density lipoprotein-cholesterol also decreased significantly from 21.4% at 0-week to 8.8 after 24 weeks (P < 0.025). On the other hand, the prevalence of hypercholesterolaemia and hypertriglycerilaemia were not significantly changed between 0 and 24 weeks (P > 0.05). Patients with DL despite treatment were characterised by higher FBG at 24 weeks of treatment compared with normolipidaemic patients (P < 0.001). It is concluded from this study that improved glycaemic control reduced some dyslipidaemia, and may therefore suffice to correct them in some Nigerian patients with NIDDM. PMID- 10821084 TI - Determination of diagnostic Widal titres in Kumasi, Ghana. AB - Three hundred and seven healthy food handlers and 34 blood-culture positive enteric fever patients were screened for Salmonellae agglutinins using the Widal test. Of the 307 healthy food handlers, only 3 (1.0%) had an anti-O titre of > or = 1/160 and 8 (2.6%) an anti-H titre of > or = 1/320 for Salmonella typhi, but the majority, 214 (69.7%) and 149 (48.5%) had titres of < 1/20 for O and H agglutinins respectively. Similar agglutinin titres were also seen for S. Paratyphi A, B, and C. In the 34 enteric fever patients, for S typhi, based on anti-O titre of > or = 1/160, 25 persons showed a significant titre, a sensitivity of 73.5%, and a specificity of 99.0%. And 21 persons showed a significant titre of > or = 1/320 for anti-H, a sensitivity of 61.8% and a specificity of 97.4%. Based on these findings, titres of > or = 1/160 and > or = 1/320 for anti-0 and anti-H respectively, were considered diagnostic for enteric fever in Kumasi, Ghana. PMID- 10821085 TI - Intestinal atresia and stenosis: a retrospective analysis of presentation, morbidity and mortality in Zaria, Nigeria. AB - Over a period of 19 years 22 children with intestinal atresia and stenosis were managed at the Ahmadu Bello University Teaching Hospital Zaria. The condition ranks as the fourth most common cause of neonatal intestinal obstruction after anorectal malformation, Hirschsprung's disease and strangulated inguinal hernia at the hospital. Three patients had duodenal atresia, 18 had jejunoileal involvement (atresia 15, stenosis 3) and one colonic atresia. The median age at presentation to the surgeon was 6 days (range 1 day-2 years). The common features were bilious vomiting and abdominal distension, the latter being more common in patients with lower atresia. Non-passage of meconium in the first 24 hours of birth occurred in 11 patients; mostly in those with lower jejunal, ileal and colonic involvement. Eight patients had associated anomalies, 5 of them with ileal atresia or stenosis. Diagnostic investigation was mainly plain abdominal radiography showing double-bubble shadow in duodenal atresia and varying degrees of air-fluid levels in other atresias. Contrast studies were not employed for diagnosis in any patient. The patients with duodenal atresia and had duodenoduodenostomy and duodenojejunostomy respectively while the jejunoileal atresia and stenosis were treated by resection and anastomosis. One patient with ileal atresia associated with total colonic Hirschsprung's disease and extensive atresia from terminal ileum down to descending colon had ileostomy. Postoperatively, 7 patients had infections of varying degrees and 3 anastomotic dehiscence. Mortality was 9 (41%) due mostly to septic complications and prematurity. While the survival of children with intestinal atresia and stenosis has improved over the years in developed countries, ours is still low (59%) due to late presentation and lack of neonatal intensive care facilities. PMID- 10821086 TI - Current status of traditional mental health practice in Ilorin Emirate Council area, Kwara State, Nigeria. AB - Twenty-seven traditional mental health practitioners (TMHPs) and 16 patients' relatives (PR) were studied with a view to gaining an understanding of the current status of traditional mental health practice in five local government areas in Ilorin Emirate Council Area, Kwara State, Nigeria. Data was collected using Practitioners' Questionnaire (PQ), Patients' Relatives' Questionnaire (PRQ), Focus Group Discussions (FGDs) and observation of TMHPs in their clinics. Factors which affect utilization of traditional mental health services were also reviewed. We found that TMHPs still enjoy considerable patronage from the populace, are more in numerical strength, and are more widely and evenly dispersed in the community than orthodox mental health practitioners (OMHPs). About 74% of TMHPs expressed interest in attending seminars aimed at improving their skills. Most of the patients' relatives expressed the belief that only traditional healers can understand the supernatural aetiological basis of mental disorders, and can therefore offer more effective care than OMHPs. Some of the negative practices observed were (i) infliction of corporal punishment and physical restraints on patients by some TMHPs resulting in wounds, which often become septic (ii) low level of hygiene at the clinics and (iii) lack of adequate follow-up care. In conclusion, since TMHPs still play a major role in the treatment of the mentally ill in this environment, OMHPs should assist them in improving on some of the negative practices identified. Thus, there is an urgent need to organize a training programme for TMHPs to expose them to the general rules of hygiene in medical care, basic principles of orthodox mental health practice, including human treatment of the mentally ill. PMID- 10821087 TI - Childhood acute lymphoblastic leukaemia (ALL) in Ghanaians and Germans--a comparative study. AB - The records of 20 ALL patients aged 12 years and below seen at Korle-Bu Teaching Hospital(KBTH) between December 1982 and May 1990 were examined as well as that of their counterparts in Hamburg University Children's Hospital(HHOG) matched by age, sex and as closely as possible date of presentation. Data on clinical and laboratory findings, treatment and outcomes were taken and analysed using binomial proportions and paired samples T-tests. It was found that KBTH patients presented with lower haemoglobin levels, bigger organ enlargements, received less intensive treatment and defaulted more often than HHOG patients. White blood cell(WBC) counts were statistically the same. Achievability of remission was 75% and 100% for KBTH and HHOG patients respectively. Average follow up was 6 months for KBTH patients and 69 months for HHOG. 60% of KBTH patients had defaulted at the time of the study. 75% of HHOG patients were alive at the time of study with the longest follow up being 114 months. It is concluded that in spite of the bigger organ enlargement for the KBTH patients, WBC count which is the single most important prognostic factor was the same for both groups, and with less intensive therapy, achievability of remission for KBTH was 75%--reasonably good. More work need to be done in Africa on prognostic features and treatment. PMID- 10821088 TI - Leucocyte counts in children with sickle cell anaemia usefulness of stable state values during infections. AB - The total white blood cell count and total and differential neutrophil counts were compared in two hundred steady state sickle cell anaemia children and sixty age and sex matched AA genotype controls to establish steady state cell counts. Steady state patients were found to have significantly higher mean total white blood cell counts (T-WBC--12.4 +/- 3.0 x 10(9/1)), mean total polymorphonuclear neutrophil counts (T-PMN--4.2 +/- 1.2 x 10(9/1)), mean total segmented (T-S--2.7 +/- 0.9 x 10(9/1)), and mean total non-segmented neutrophil counts (T-NS--1.5 +/- 0.5 x 10(9/1)), when compared with their M genotype controls; T-WBC (5.2 +/- 1.9 x 10(9/1)) (P < 0.001), T-PMN (2.2 +/- 8.0 x 10(9/1)) (P < 0.001), T-S (2.2 +/- 0.9 x 10(9/1)) (P < 0.01), and T-NS (0.02 +/- 0.04 x 10(9/1)) (P < 0.001). Contrary to an expected neutrophilia, steady state patients had a low percentage polymorphonuclear neutrophil count (34%) which did not change significantly with increasing age (P > 0.05). The raised counts in steady state patients suggest that previously established criteria for the diagnosis of bacterial infections cannot be applied to sickle cell patients, and thus new criteria must be generated. PMID- 10821089 TI - Characteristics of VVF patients as seen at the specialist hospital Sokoto, Nigeria. AB - This paper is a preliminary report of a longitudinal study of VVF patients seen at the specialist hospital Sokoto, Nigeria. Thirty-one (31) patients admitted into the VVF ward during the period, May 1996 to April 1997 were studied. The patients were mainly Hausa/Fulani Muslims (100%), short-statured (mean height, 149 cm), full-time house wives (100%) and were married at the mean age of 13 years. Only 6% of the patients could read or write in English while 81% had Quranic school education only. 60% of the women live in rural communities which lacked, or remotely located to appropriate health care services. Most of the cases developed the disease at the mean age of 15 years. Majority of the cases (77%) did not receive antenatal care while the mean duration of labour for all the patients was 4 days. 84% of the patients were eventually referred to hospital for delivery. Pregnancy wastage rate was 87%. Averagely, twenty-six, (84%) of the patients have had 3 surgical repairs done. Divorce rate following the occurrence of the disease was 55%. Our recommendations include: the integration of maternal and child health (MCH) education in the curriculum of Quranic schools, and advocacy for free antenatal care and delivery services at the primary and secondary levels of health care, special health education programs on maternal health for religious leaders, training of more personnel, and bringing MCH services closer to the grassroots. PMID- 10821090 TI - How often are Ghananian gallbladder stones cholesterol-rich. AB - BACKGROUND: Although the prevalence of symptomatic gallbladder stone disease in Ghana increased almost four-fold between 1966 and 1991, little is known about the composition of and aetiopathogenesis of these stones, nor about their suitability for non-surgical (dissolution) treatments. METHODS: To study this, gallstones from 67 out of 90 patients coming to cholecystectomy were retrieved and, based on their external appearance, classified provisionally as cholesterol(chol) (n = 8), black pigment (n = 28) and brown pigment (n = 31) stones. The gallstones were then homogenised, their cholesterol(chol) content measured chemically and the stones re-classified as cholesterol-poor (< 10% chol by weight), intermediate (10 75% chol) and cholesterol-rich (> 75% chol). The relationship between the initial and the definitive classifications was then examined and the biliary bacteriology (carried out on fresh samples of gallbladder(GB) bile obtained by fine needle aspiration) on gallstone composition, analysed. RESULTS: The external appearance correctly predicted stone composition in the 28 thought, initially, to have black pigment stones (all of whom had stones containing < 10% chol by weight, on chemical analysis) and the eight believed, originally to have "cholesterol" stones (all of whom had stones with > 75% chol) but it proved unreliable in the 31 considered, at the time of surgery, to have brown pigment stones (mean chol content 58+ SEM 35%; range 0-98%. By chemical analysis, more than half the patients 35 of 67 or 52% had cholesterol-poor stones, nine (13% of the total) had intermediate stones, while 23 (34%) had cholesterol-rich stones. Cholesterol-rich gallstones were also more frequent in women than in men (p < 0.03). Only nine of 43 patients (21%) whose GB bile was aspirated, had positive bacterial cultures. There was no obvious difference in stone composition between those with positive, and those with negative, cultures. CONCLUSION: Since the majority of Ghanaian patients with cholecystolithiasis have gallbladder stones with < 75% chol by weight, when active treatment is indicated surgery is more appropriate than dissolution therapy. However contrary to common belief, cholesterol-rich gallstones do occur in West Africa: 34% of the present series had stones with > 70% chol by weight. PMID- 10821091 TI - Chemical injuries to the eye in Benin City, Nigeria. AB - Chemical injury to the eye is still an important cause of blindness and serious complications in Benin City, Nigeria. Twelve patients were seen with chemical injury to the eye over a five year period. They were all male patients. Ten were assault cases and two were accidental cases. The most common injurious agent was ammonia. Complications seen were corneal opacification in ten patients, symblepharon in nine patients, entropion, ectropion, cataract and staphyloma developed in some of the patients. There were four cases of bilateral blindness and six cases of unilateral blindness. Late presentation to hospital and failure to initiate adequate first aid treatment were contributing factors to the poor outcome. Health education of the public about the importance of prompt, prolonged irrigation of the chemically injured eye and early presentation to the hospital is advised. PMID- 10821092 TI - Heterotopic pregnancy following ovulation stimulation with clomiphene: a report of three cases. AB - Heterotopic pregnancy was a medical rarity, prior to the use of ovulation inducing drugs and advent of assisted reproduction technology. Three cases were encountered in the course of clinical practice in a teaching hospital, over a period of six and a half years. Two of them occurred in association with ovulation induction with CLOMIPHENE and the third case with no identifiable underlying cause. We postulate dizygotic twinning, highest among the Yoruba race of Nigeria as a possible aetiological factor. One of the two cases following CLOMIPHENE has been highlighted in more details, together with a review of literature. Diagnosis in all cases was made in retrospect following surgical intervention for the ruptured ectopic components. All the patients presented with persistent pregnancy symptoms, with sonographic confirmation of viable intrauterine fetuses which were carried to term. PMID- 10821093 TI - Balloon dilation and stenting of chronic iliac vein obstruction: technical aspects and early clinical outcome. AB - PURPOSE: To describe the technical aspects of percutaneous balloon dilation and stenting for the treatment of venous outflow obstruction in chronic venous insufficiency. METHODS: Between March 1997 and December 1998, 94 consecutive patients (median age 48 years, range 14 to 80) with suspected iliac vein obstruction in 102 limbs were studied prospectively with the intent to treat any venous occlusion or stenosis verified during femoral vein cannulation. Data from the history, clinical examination, procedure, and follow-up were recorded. Preoperative indicators of obstruction were venographic evidence of occlusion, stenosis, or pelvic collateral vessels; increased arm-foot venous pressure differential; and abnormal hyperemia-induced venous pressure elevation. RESULTS: Cannulation and technical success rates were 98% and 97%, respectively, with 118 Wallstents deployed in 77 veins. Primary, assisted primary, and secondary patency rates at 1 year were 82%, 91%, and 92%, respectively. Clinical improvement in pain and swelling was significant. CONCLUSIONS: Stenting of benign iliac vein obstruction is a safe method with good short-term results. Venous lesions should always be stented; when treating iliocaval junction lesions, stents should be inserted well into the inferior vena cava. Absence of collateral vessels does not exclude the existence of significant obstruction, and their presence may indicate an obstruction not visualized. No gold standard for accurate pre- or intraoperative patient selection is currently available. PMID- 10821094 TI - A major step forward in the treatment of venous occlusion. PMID- 10821095 TI - Matched-pair analysis of conventional versus endoluminal AAA treatment outcomes during the initial phase of an aortic endografting program. AB - PURPOSE: To investigate whether endovascular stent-grafts implanted during the early phase of an aortic endografting program have advantages over conventional surgical procedures for treatment of infrarenal aortic aneurysm (AAA). METHODS: In the first months of an endografting program, 37 patients (36 men; mean age 67.9 +/- 7.1 years, range 55 to 86) underwent AAA repair with endovascular implantation of a Vanguard (n = 17) or Talent (n = 20) bifurcated stent-graft. Data collected during the perioperative period and in follow-up were compared retrospectively to a matched group of 37 elective surgical patients. RESULTS: All endograft implantations were completed. Two type I and 6 type II endoleaks (21.6%) were seen postoperatively. Five type II sealed without intervention; 1 type I endoleak was corrected with an additional stent, but 1 type I and 1 type II endoleaks persisted despite attempts with coil embolization. Two (5.4%) endograft patients died during the perioperative period; however, this was not significantly different (p = 0.15) from the control group. In the mean follow-up of 12 +/- 6 months for both groups, 1 (2.7%) late conversion was necessary at 2 years for aneurysm expansion in an endograft patient with an unsealed type I endoleak. CONCLUSIONS: In our learning curve experience with aortic endografting, postoperative morbidity and mortality were higher in endograft patients compared to conventionally treated controls. Only in the endograft group was reoperation required during follow-up. Careful monitoring with periodic imaging studies is mandatory after endoluminal AAA treatment. PMID- 10821096 TI - How will endoluminal grafting for aneurysms be judged? PMID- 10821097 TI - First-generation aortic endografts: analysis of explanted Stentor devices from the EUROSTAR Registry. AB - PURPOSE: To examine the structure and healing characteristics of chronically implanted Stentor endografts that were explanted due to migration, endoleak, thrombosis, or aneurysm expansion. METHODS: The devices were harvested following reoperation (n = 5) or autopsy (n = 1) with implantation times ranging from 13 to 53 months. Structural modifications to the metal components were examined using radiography, endoscopy, and magnetic resonance imaging (MRI). Specimens taken from components of the modular stent-grafts were examined histologically and with scanning electron microscopy (SEM) to assess healing behavior. Physical and chemical stability of the nitinol wires and woven polyester graft material was evaluated using SEM and electron spectroscopy for chemical analysis. RESULTS: Although the endografts were retrieved for a variety of reasons, they exhibited similar healing and structural modifications. The woven polyester sleeve showed evidence of yarn shifting and distortion, yarn damage, and filament breakage leading to the formation of openings in the fabric. The luminal surface endografts showed incomplete healing characterized by a poorly organized, nonadherent thrombotic matrix of variable thickness. Radiographic and endoscopic observations indicated that structural failure of the grafts, particularly in the main aortic component, was related to severe compaction and dislocation of the metallic frame due to suture breaks. Corrosion marks were observed on some nitinol wires in all devices. Chemical analysis and ion bombardment of the nitinol wires revealed that the surface concentrations of titanium and nickel were not homogenous. The first layer was composed of carbon or organic elements, followed by a stratum of highly oxidized titanium with a low nickel concentration; the titanium-nickel alloy lay beneath these layers. CONCLUSIONS: Although the materials selected for construction of endovascular grafts appears judicious, the assembly of these biomaterials into various interrelated structures within the device requires further improvement. PMID- 10821098 TI - Malpositioned or dislocated aortic endoprostheses: repositioning using percutaneous pull-down maneuvers. AB - PURPOSE: To present the capabilities and potential complications of 2 percutaneous techniques for repositioning malpositioned or dislodged aortic endografts. METHODS: Seven male patients (median age 67.9 years, range 59 to 78) required correction of misplaced or dislocated endografts in the thoracic (n = 1) or infrarenal abdominal aorta (n = 6). In 1 patient, an infrarenal bifurcated stent-graft was mistakenly deployed across a renal artery; repositioning was accomplished by tugging caudally on a guidewire placed across the endograft bifurcation and exteriorized from both femoral arteries. An inflated balloon catheter was used to reposition 3 dislocated aortic devices (1 thoracic, 2 infrarenal) and 3 iliac graft limbs that had disconnected from the main graft body 6 to 12 months after implantation. RESULTS: Repositioning maneuvers were successful in all cases, with the devices being moved from 5 to 27 mm (median 7.8 mm). There were no procedure-related complications. CONCLUSIONS: Nonsurgical repositioning of misplaced aortic prostheses is technically feasible in individual cases. The risk associated with the procedure, however, cannot yet be evaluated. PMID- 10821099 TI - Cerebrospinal fluid drainage to reverse paraplegia after endovascular thoracic aortic aneurysm repair. AB - PURPOSE: To report a case of endovascular descending thoracic aortic aneurysm (TAA) repair in which delayed-onset paraplegia was reversed using cerebrospinal fluid (CSF) drainage. METHODS AND RESULTS: A 74-year-old patient with a 6.0-cm TAA underwent endovascular stent-graft repair that involved overlapping placement of 3 Talent devices to cover the 31-cm-long defect. Twelve hours later, a neurological deficit occurred manifesting as left leg paralysis with paresis on the right. After urgent intrathecal catheter placement and drainage of cerebrospinal fluid for 48 hours, the neurological deficit resolved. The patient's clinical condition was normal and endoluminal exclusion of the TAA remained secure at 8-month follow-up. CONCLUSIONS: This case demonstrates the potential therapeutic role for CSF drainage to reduce the complications of spinal cord injury after endovascular thoracic aneurysm repair. PMID- 10821100 TI - Stent-graft treatment of a common carotid artery pseudoaneurysm. AB - PURPOSE: To report a case demonstrating successful endovascular treatment of a right common carotid artery pseudoaneurysm using a commercially prepared balloon expandable covered stent. METHODS AND RESULTS: A 50-year-old man was evaluated for syncopal episodes. He had a history of severe trauma sustained in a motor vehicle accident 3 years before symptom onset. Doppler ultrasound scanning detected a pseudoaneurysm at the origin of the right common carotid artery. The defect measured 25 mm x 20 mm with a 22-mm-long neck on angiography and computed tomography; there was no evidence of carotid stenosis or associated vascular pathology. Via a percutaneous femoral access, 2 Jostent peripheral stent-grafts were placed at the level of the aneurysm, safely achieving complete repair of the arterial wall defect. The patient was asymptomatic at his 12-month evaluation. Color flow duplex scans showed continued exclusion of the pseudoaneurysm. CONCLUSIONS: Wide-necked aneurysms in the extracranial carotid arteries may be treated with stent-grafts, which can achieve complete and permanent reconstruction of the arterial wall by excluding the aneurysm. PMID- 10821102 TI - Coronary techniques and devices: potential applications in peripheral vessels. PMID- 10821101 TI - Vibrational angioplasty and hydrophilic guidewires in the treatment of chronic total coronary occlusions. AB - PURPOSE: To study the efficacy and safety of vibrational angioplasty versus hydrophilic guidewires for recanalizing chronic total occlusions (CTOs) of the coronary arteries. METHODS: In a retrospective study, 99 patients with old (>3 months) CTOs resistant to conventional techniques were treated either with vibrational angioplasty (group A, n = 72) or 0.014-inch hydrophilic guidewires (group B, n = 27). The selection of the technique (vibrational angioplasty or hydrophylic guidewires) was dependent only upon device availability. A variety of guidewires were employed in conjunction with vibrational angioplasty. RESULTS: The crossing success rates in groups A and B were 86.1% (62/72) and 55.5% (15/27) (p < 0.05), with final procedural success rates of 75% (54/72) and 44.4% (12/27) (p < 0.01), respectively. The main reasons for failure were inability to cross the lesion with a guidewire (10/18 in group A and 12/15 in group B) and large dissections resulting in vessel closure (2/18 in group A and 2/15 in group B). Three major complications were seen, 2 in group A and 1 in group B. Three vessel perforations were reported in group A. Both techniques needed prolonged fluoroscopy times. CONCLUSIONS: Vibrational angioplasty was more successful in treating CTOs compared to hydrophilic guidewires and had similar complication rates (most without clinical sequelae). PMID- 10821103 TI - Osteomyelitis of the spine and abscess formation in the left thigh after stent graft implantation in the superficial femoral artery. AB - PURPOSE: To present a rare case of abscess formation around a covered stent in the superficial femoral artery. METHODS AND RESULTS: Two weeks after balloon dilation of a left superficial femoral artery (SFA) occlusion, during which a Hemobahn covered stent had been placed to treat dissection, a 77-year-old nondiabetic male developed intolerable pain and swelling of his left thigh. An abscess had formed around the stent, which was patent; intravenous antibiotic therapy quelled the symptoms, and the patient discontinued his oral antibiotic regimen weeks after discharge. General septicemia ensued. Acute lower limb ischemia and excruciating back pain prompted readmission. The SFA stent-graft occlusion required femoropopliteal bypass; the abscess and spondylodiskitis that had developed in the T12 and L1 vertebrae responded to intravenous antibiotics. The patient is without signs of infection at 6 months. CONCLUSIONS: Local and systemic infections associated with intraluminal prostheses are rare, and prophylactic antibiotic therapy is not commonly employed. Balloon- or device induced arterial injury may expose the arterial wall to bacterial colonization, suggesting that patients receiving lengthy stents or experiencing arterial injury during angioplasty should receive antibiotics as a precautionary measure. PMID- 10821104 TI - Endovascular treatment of multiple visceral artery paradoxical emboli with mechanical and pharmacological thrombolysis. AB - PURPOSE: To report a case of paradoxical emboli to multiple visceral vessels treated with both mechanical (AngioJet device) and pharmacological (urokinase) thrombolysis. METHODS AND RESULTS: A 72-year-old man presented with a 48-hour history of symptomatic right renal ischemia, which was treated with heparinization. Five days later, an abrupt creatinine elevation prompted arteriography, which demonstrated thromboembolism of the superior mesenteric artery (SMA) and both renal arteries. The AngioJet aspiration device was employed to successfully remove the clot from the SMA; urokinase infusion restored flow to the left kidney. At the 16-month follow-up evaluation, the patient was normotensive without medication and had a stable creatinine (1.4 mg/dL). CONCLUSIONS: Because of its speed and minimal morbidity, the AngioJet device may be an attractive alternative to surgical embolectomy or pharmacological thrombolysis in highly selected cases of acute visceral artery thromboembolism. PMID- 10821105 TI - Symptomatic subtotal occlusion of the innominate artery treated with balloon angioplasty and stenting. AB - PURPOSE: To report the endovascular treatment of a subtotal occlusion of the innominate artery giving rise to subclavian steal syndrome. METHODS AND RESULTS: A 60-year-old man in general good health was admitted to the hospital for sudden onset of amaurosis in the right eye. Thrombosis of the central retinal artery was diagnosed. Physical examination, color flow duplex imaging, and aortic arch angiography showed a subtotal occlusion of the innominate artery with right subclavian steal syndrome. One month later, balloon dilation and stenting of the innominate artery was performed through a right axillary access without cerebral protection. The innominate artery was recanalized with correction of the steal syndrome and restoration of the right radial pulse; no complications occurred. Twelve months later, color flow duplex sonography confirmed innominate stent patency and antegrade flow in the right vertebral artery. CONCLUSIONS: Our experience supports the view that percutaneous endovascular techniques are appropriate and are the preferred treatment for lesions of the supra-aortic vessels. Continued surveillance will determine their long-term durability. PMID- 10821106 TI - Regarding outpatient carotid artery stenting. PMID- 10821107 TI - Alkaloids from frog skin: the discovery of epibatidine and the potential for developing novel non-opioid analgesics. PMID- 10821108 TI - Recent studies of the mechanism of protein prenylation. PMID- 10821109 TI - Chiroptical methods in the stereochemical analysis of natural products. PMID- 10821110 TI - Biosynthesis of pyrrolnitrin and other phenylpyrrole derivatives by bacteria. PMID- 10821111 TI - Diterpenoids. PMID- 10821112 TI - Simple indole alkaloids and those with a nonrearranged monoterpenoid unit. PMID- 10821113 TI - Oligomeric proanthocyanidins: naturally occurring O-heterocycles. PMID- 10821114 TI - Apoptotic response and cell cycle transition in ataxia telangiectasia cells exposed to oxidative stress. AB - The recently identified ATM gene plays a role in a signal transduction network activating multiple cellular functions in response to DNA damage. An attractive hypothesis is that the ATM protein is involved in a specialized antioxidant system responsible for detoxifying reactive oxygen intermediate and that the absence or dysfunction of this protein in AT cells would render them less capable of dealing with oxidative stress. In order to investigate the role of the ATM gene in cell cycle control and programmed cell death, Lymphoblastoid cell lines derived from four Ataxia-Telangiectasia (AT) patients and six controls have been analyzed. All cell lines were incubated with 2-deoxy-D-ribose (dRib), a reducing sugar that induces apoptosis through oxidative stress. The result showed an impaired response to dRib-induced apoptosis in AT cells, as well as a defect of cellular cycle arrest in G1/S phase and a normal expression of p53 protein. This indicate that the kinase activity of ATM gene product plays a very important role in the cellular response to oxidative stress. In conclusion the altered response of AT cells to oxidative stress and particularly their resistance to apoptotic cell death, could explain the high predisposition of these cells to progress toward malignant transformation. PMID- 10821115 TI - The effect of selenium and/or vitamin E treatments on radiation-induced intestinal injury in rats. AB - Cytotoxic effects of ionizing radiation on gastrointestinal epithelium may be related to oxidative stress. In this study, we wanted to investigate the effects of selenium, vitamin E and selenium plus vitamin E pretreatments prior to whole abdominal irradiation on intestinal injury. Irradiation caused increased lipid peroxide and decreased GSH levels in the intestine. Intestinal superoxide dismutase and glutathione peroxidase activities were increased, but glutathione transferase activity decreased following irradiation. Selenium and/or vitamin E pretreatments ameliorated these disturbances in prooxidant-antioxidant balance. This amelioriation has been verified with histopathological findings. These results indicate that antioxidant pretreatments prior to irradiation may have some beneficial effects against irradiation-induced intestinal injury. PMID- 10821116 TI - Opioid propeptide mRNA content and receptor density in the brains of AA and ANA rats. AB - Recent evidence has indicated an association between the rewarding effects of ethanol intake and endogenous opioid activity. The present studies examine the presence of differences in opioid peptide mRNA content and mu and kappa opioid receptor densities, between ethanol naive AA and ANA rats bred selectively for their high and low alcohol consumption, respectively. In situ hybridization was used to compare the content of proopiomelanocortin, proenkephalin and prodynorphin mRNA in distinct brain regions known to be involved in the reinforcing properties of addictive drugs, between rats from each line. Results indicated that AA rats had a significantly greater content of proopiomelanocortin mRNA in the arcuate nucleus of the hypothalamus, of proenkephalin mRNA in the prefrontal cortex and of prodynorphin mRNA in the mediodorsal nucleus of the thalamus (p < or = .05). Receptor autoradiography was performed using 3H-labeled ligands specific for mu and kappa opioid receptors. AA rats were found to have a greater density of mu opioid receptors in the shell region of the nucleus accumbens and prefrontal cortex, but a lower density of kappa opioid receptors in the ventromedial hypothalamus, compared to ANA rats. The present data demonstrate the presence of inherited differences in the activity of distinct components of the endogenous opioid system in some brain regions associated with the processes of reward and reinforcement; and as such, may play a role in determining differences in ethanol drinking between AA and ANA rats. PMID- 10821117 TI - The effect of growth hormone on the proliferation of human Th cell clones. AB - The effects of human growth hormone (hGH) on human Th cell clones were examined. Both 20K and 22K hGH stimulated the proliferation of Th2 and Th0 cells in the presence of mite antigen, whereas they did not stimulate the proliferation of Thl cells. Because the effect of 20K hGH was almost the same as that of 22KhGH, it was suggested that the action of hGH was not mediated through prolactin receptor but through hGH receptors. The application of growth hormone binding protein (GHBP) inhibited the cell growth of Th1 clones. In Th2 and Th0 cells GHBP inhibited the hGH-stimulated cell proliferation. However, GHBP alone did not affect the proliferation of Th2 and Th0 cells. hGH was detected in the supernatant of Th1 clones in the presence of mite antigen but it was not detected in Th2 clones. hGH was detected in one out of 4 batches of Th0 clones. These data indicated that hGH was secreted from Thl clones, and that Th0 clones possessed characteristics of both Th2 and Th0 clones. PMID- 10821118 TI - Role of chronic exercise in decreasing oxidized LDL-potentiated platelet activation by enhancing platelet-derived no release and bioactivity in rats. AB - This study investigates how chronic exercise affects Ox-LDL mediated-platelet activation. Five-week-old male Wistar rats were assigned to either control or trained groups. Trained rats were treadmill-trained for 10 weeks after familiarization. The following measurements were taken in both control and trained groups: plasma lipid profile, oxidation of LDL, platelet adhesiveness, aggregability, cGMP contents, plasma and platelet-NO metabolite (nitrite plus nitrate) levels, and urinary 8-iso-prostaglandin F2alpha (8-iso-PG F2alpha) levels. Based on those measurements, major findings in this study can be summarized as follows: 1) the trained group prolonged the lag time of isolated LDL subjected to copper-induced in vitro oxidation significantly longer than the control group; 2) although having higher plasma and platelet derived-NO metabolite levels, the trained group had lower urinary excretion of 8-iso PGF2alpha than the control group; 3) the trained group had a lower platelet adhesiveness and aggregability and higher platelet derived-NO metabolite and cGMP productions than the control group; 4) the trained group had a lower Ox-LDL potentiated platelet adhesiveness and aggregability and Ox-LDL-attenuated NO metabolite and cGMP productions in platelet than the control group; and 5) treating the platelet with L-arginine inhibited Ox-LDL-potentiated platelet activation in both control and trained groups. Results in this study demonstrate that amounts of preformed lipid peroxides decrease while NO production (which acts as an antioxidant) is significantly increased after chronic exercise. Moreover, exercise training decreases Ox-LDL-potentiated platelet activation most likely by enhancing platelet-derived NO release and bioactivity. PMID- 10821119 TI - Metabolic defects caused by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and by HPTP (the tetrahydropyridinyl analog of haloperidol), in rats. AB - The results of previous studies in the baboon have suggested that HPTP, the tetrahydropyridinyl analog of haloperidol causes a urinary biochemical marker profile similar to those seen in humans suffering from inborn errors of mitochondrial respiration. In order to identify a possible relationship between compromised cellular energy production and neuronal damage we now have compared the urinary profiles of rats treated with the pro-neurotoxin, MPTP as well as with HPTP. Significantly increased urinary excretion of lactic acid and 2 ethylhydracrylic acid in MPTP and HPTP treated rats was observed, indicating that both MPTP and HPTP and/or their respective metabolites cause mitochondrial inhibition in the rat. PMID- 10821120 TI - Establishment of Salmonella strain expressing catalytically active human UDP glucuronosyltransferase 1A1 (UGT1A1). AB - Human uridinediphosphate-glucuronosyltransferase 1A1 (UGT1A1) was expressed in Salmonella typhimurium TA1535 cells by transfection of the cells with plasmids carrying the UGT1A1 cDNA. UGT1A1 cDNA was isolated by a polymerase chain reaction from human liver total RNA and was inserted into the pSE420 plasmid, linked to the trc promoter and terminator. The plasmid thus constructed was introduced into Salmonella TA1535 cells. The expression of human UGT1A1 protein was confirmed by Western blot analysis. The maximal expression was observed at 24 h after the addition of isopropyl-beta-D-thiogalactopyranoside, an inducer. However, the bilirubin conjugation activity of the membrane fraction from the Salmonella cells was not detectable. When a beta-glucuronidase inhibitor such as saccharic acid 1,4-lactone, glycyrrhizin or 1-naphtyl-beta-D-glucuronide was added to the reaction mixture, the bilirubin conjugation activity of the human UGT1A1 was detected. When geniposide was added to the reaction mixture, the bilirubin conjugation activity of UGT1A1 was not seen. Taking these results into account, the established Salmonella strain possesses the beta-glucuronidase activity. Since the beta-glucuronidase activity of the Salmonella was lower than that of E. coli, it was concluded that Salmonella seemed to be a good host to express UGT protein. This is the first study to demonstrate the establishment of a bacterial strain expressing native human UGT protein showing catalytic activity. PMID- 10821121 TI - Age-dependent effect of Freund's adjuvant on 24-hour rhythms in plasma prolactin, growth hormone, thyrotropin, insulin, follicle-stimulating hormone, luteinizing hormone and testosterone in rats. AB - The effect of Freund's adjuvant administration on 24-hour changes of plasma prolactin, growth hormone (GH), thyrotropin (TSH), insulin, follicle-stimulating hormone (FSH), luteinizing hormone (LH) and testosterone were studied in young (2 months) and aged (18 months) male Wistar rats. Rats were injected s.c. with Freund's adjuvant or adjuvant's vehicle and, 18 days later, they were killed at 6 different time intervals throughout a 24-hour cycle to measure circulating hormone levels by specific RIAs. Young rats receiving adjuvant's vehicle exhibited significant time-of-day-dependent variations in plasma TSH, LH and testosterone, with maximal levels at 1300 h, 0100 h and 1700 h, respectively. Prolactin and insulin levels, analyzed globally in a factorial ANOVA, showed significant time-of-day changes with maximal levels at 1300 - 1700 h and 2100 h, respectively. The daily rhythms in plasma LH and testosterone found in young rats were not longer observed in Freund's adjuvant-injected rats, while as far as TSH, a second peak was observed at 0100 h after Freund's adjuvant administration. Twenty-four hour rhythms in circulating TSH, LH and testosterone were blunted in old rats receiving either Freund's adjuvant or its vehicle. Aged rats exhibited significantly higher circulating levels of prolactin, and lower levels of GH, TSH, FSH and testosterone. The results indicate that secretion of prolactin, GH, TSH, FSH and testosterone are age-dependent, as are the responses of TSH, LH and testosterone to Freund's adjuvant administration. PMID- 10821122 TI - Sigma ligands inhibit the growth of small cell lung cancer cells. AB - The effects of sigma ligands on small cell lung cancer (SCLC) cells were investigated. 125I-N-(2-(piperidino)ethyl)-2-iodobenazmide (2-IBP) bound with high affinity to SCLC cell line NCI-H209 and NCI-N417. Specific 125I-2-IBP binding was inhibited with high affinity by ifendipine, haloperidol, (2 piperidinyl-aminoethyl)-4-iodobenzamide (IPAB) and 1,3-ditolylguanidine (DTG) with IC50 values of 3, 10, 15 and 90 nM respectively. In vitro, 10 microM 2-IBP, haloperidol or IPAB inhibited NCI-N417 proliferation using a MTT or clonogenic assay. In vivo, 4 mg/kg IPAB or 2-IBP inhibited NCI-N417 xenograft proliferation. 125I-2-IBP localized to the SCLC tumors after subcutaneous injection. These results suggest that sigma ligands may be utilized to localize and inhibit the proliferation of SCLC tumors. PMID- 10821123 TI - Influence of acetylsalicylic acid on oxidation of native and glycated low-density lipoprotein. AB - It is generally accepted that oxidation of low-density lipoproteins (LDL) is a causal factor in the development of atherosclerosis. Non-enzymatic glycosylation of LDL, i.e."glycation", plays a central role in late complications of diabetes mellitus and may initiate and/or accelerate the oxidation process. Therefore, the inhibition of this processes is of major therapeutic relevance. The influence of acetylsalicylic acid (ASA) on the oxidation of native and glycated LDL was studied in vitro. LDL (0.25 mg protein/ml ) was oxidatively modified with 5.0 microM CuSO4. Only at "supratherapeutical" ASA concentrations in the range 0.06 2.0 mg /ml we found a significant concentration-dependent inhibition of LDL oxidation both for native and glycated LDL, which was from 0.2 mg/ml upwards significantly more marked for native LDL than for glycated LDL. The maximal inhibitory effect occurred at 2.0 mg/ml with 89.6% inhibition of LDL-oxidation for native LDL and 64.4% for glycated LDL. At 0.2 mg/ml ASA the respective inhibitory values were 38.5% and 31.0%. For glycated LDL the ASA doses of maximal and approximately 50%-inhibition, as found for native LDL, were chosen to investigate the inhibitory effect on 2,4,8 and 24 hours oxidation of glycated LDL to monitor the time-dependency of inhibition by ASA. This revealed that ASA only delayed, not permanently inhibited LDL oxidation. PMID- 10821124 TI - Somatosensory cortex stimulation-evoked analgesia in rats: potentiation by NO synthase inhibition. AB - Clinical and immunohistochemical evidence suggests the possible significance of electrical stimulation of the secondary somatosensory cortex (S-II) as an analgesic therapy. The aim of the present study was to gain behavioral evidence for S-II stimulation-induced antinociception in conscious rats and to evaluate if the evoked antinociception can be potentiated by the neuronal NO synthase inhibitor 7-nitro-indazole. S-II stimulation produced a weak antinociception in the formalin-induced nociception test, but not in the thermal or mechanical nociception tests. This effect was remarkably potentiated by systemic administration of 7-nitro-indazole at a small dose that had no effect by itself. Thus, our data provide behavioral evidence for S-II stimulation-induced analgesia and may also predict a novel therapeutic strategy in combination with NO synthase inhibitors. PMID- 10821125 TI - Destabilization of tumor necrosis factor-alpha mRNA by 5-alpha dihydrotestosterone in Jurkat cells. AB - The effect of a male steroid hormone, 5DHT, on the expression of TNF-alpha was examined using a human leukemia T cell line, Jurkat. Cells were treated with 5DHT in the presence or absence of PHA, and RNA was isolated followed by a reverse transcriptase - mediated PCR (RT-PCR) to measure the steady state levels of TNF alpha mRNA. The treatment of cells with 5DHT resulted in a 50% of decrease in the level of TNF-alpha mRNA compared to that in untreated conditions (basal level). A similar level of reduction of the message by 5DHT was also observed in PHA stimulated cells. The reduction of the steady state levels of TNF-alpha mRNA in Jurkat cells was a result of destabilization of the gene as demonstrated by actinomycin D treatment; a half-life of TNF-alpha message in 5DHT treated cells and non-treated cells was 1 hr and 2.5 hr, respectively, whereas that in 5DHT/PHA and PHA-treated cells was 3hr and 6hr, respectively. PMID- 10821126 TI - Eating habits and intensity of medication in elderly hypertensive outpatients. AB - Several dietary factors have been shown to lower blood pressure in elderly patients in clinical trials, but it is not known how eating habits affect blood pressure control in outpatients treated with antihypertensive drugs. We obtained data on dietary information regarding food groups rich in potassium, magnesium, and calcium by submitting a questionnaire to 190 elderly outpatients. Blood pressure levels and the intensity and cost of antihypertensive treatment were obtained from clinical records. The mean age and blood pressure were 72.3 +/- 9.3 years and 138.7 +/- 15.2/74.6 +/- 8.8 mmHg, respectively. Patients were divided into three groups on the basis of the intensity of medication: the LS group (n=52), treated with a low dose of a single drug; the HS group (n=95), treated with a high dose of a single drug; and the M group (n=43), treated with multiple drugs. Average age, body mass index, blood pressure levels, and extent of target organ damage were similar among the three groups. Intake frequency (rarely, once or twice per week, 3 to 6 times per week, or every day) of the food groups was compared among the three groups. The LS group ate fruit (p < 0.05) and seaweed (p < 0.01) with significant frequency compared with the other groups, whereas milk and dairy products were taken with similar frequency by all groups. The monthly cost of antihypertensive medications averaged 17,218 +/- 620 yen in the LS group, 17,746 +/- 375 yen in the HS group, and 20,066 +/- 1,364 yen in the M group. These data suggest that habitual intake of foods rich in potassium and magnesium are associated with reduced intensity and cost of medication and with preservation of blood pressure control in elderly hypertensive outpatients. PMID- 10821127 TI - Deletion polymorphism of ACE gene is associated with higher blood pressure after hospitalization in normotensive subjects. AB - Blood pressure has been shown to decrease in response to hospital admission. Several parameters including the decline of sympathetic nervous activity and negative sodium balance have been shown to be involved in this phenomenon. We investigated genetic influence on office BP and BP after hospitalization. One hundred and sixty-three men from the general population, free from antihypertensive medication, were enrolled in the present study. They stayed at the hospital for general medical check-up. BP was measured on the day of admission, and again the following day. Mean systolic blood pressure was significantly decreased after hospitalization from 117.3 +/- 9.9 mmHg to 115.3 +/ 12.8 mmHg (p=0.042). Subjects with DD+ID genotype showed a significantly higher systolic blood pressure after hospitalization than that of subjects with genotype II. There were no genotype specific differences in diastolic blood pressure or changes in blood pressure by the administration. In summary, systolic blood pressure after hospitalization was significantly higher in normotensive male subjects who possessed the D allele of ACE I/D polymorphism. PMID- 10821128 TI - Suppression of cardiac sympathetic nervous system during dental surgery in hypertensive patients. AB - We determined the changes in blood pressure, pulse rate, and heart rate variability during dental surgery in hypertensive patients. The study included 18 essential hypertensives and 18 age and sex matched normotensive controls who underwent tooth extraction at our hospital. Holter electrocardiographic monitoring was used to determine the power spectrum of R-R variability before and during dental surgery. The low frequency (LF: 0.041 to 0.140 Hz), high frequency (HF: 0.140 to 0.500 Hz), and total spectral powers (TF: 0.000 to 4.000 Hz) were calculated, and the ratio of LF to HF and the percentage of HF relative to TF (%HF: HF/TF x 100) were used as indexes of sympathetic and parasympathetic activities, respectively. The baseline blood pressure for hypertensive patients (149 +/- 4/85 +/- 2 mmHg) was significantly higher than that for normotensive patients (119 +/- 3/71 +/- 2 mmHg). The baseline pulse rates were similar between the two groups. Blood pressure increased during tooth extraction in both groups; however, changes in blood pressure did not differ between them. Administration of local anesthetic significantly decreased the %HF in normotensive patients (before vs. after anesthesia; 22.3 +/- 2.4 vs. 13.8 +/- 2.7%, p < 0.05). In contrast, the LF/HF significantly decreased during the local anesthesia and tooth extraction in hypertensive patients. These results suggest that pressor response induced by tooth extraction did not differ between normotensive and hypertensive patients, and that suppression of the cardiac sympathetic nervous system during dental surgery might attenuate the pressor response in patients with hypertension. PMID- 10821129 TI - Low-dose atropine attenuates muscle sympathetic nerve activity in healthy humans. AB - Central muscarinic receptors play an important role in the regulation of cardiac vagal nerve activity. We studied the inhibition of central muscarinic receptors and sympathetic nerve function in humans, since very little information is currently available on this subject. We examined the effects of graded doses of atropine (five doses, range 0.001 to 0.016 mg/kg) on heart rate, arterial pressure, heart rate variability, and muscle sympathetic nerve activity in 13 healthy young volunteers. Atropine caused biphasic effects on heart rate and the high-frequency (HF) power of R-R interval variability. At lower doses (< or =0.002 mg/kg for heart rate, 0.001 mg/kg for HF power), atropine decreased heart rate and increased HF power. In contrast, at higher doses, atropine increased heart rate and decreased HF power. Low-dose atropine significantly attenuated muscle sympathetic nerve activity, burst rate (bursts/min) by -30.5 +/- 6.0% and burst incidence (bursts/100 heart beats) by -23.8 +/- 6.9% at 0.002 mg/kg. Systolic and diastolic arterial pressure did not change with atropine infusion. Low-dose atropine (< or =0.002 mg/kg) did not significantly affect either low frequency (LF) power or LF/HF. These results suggest that central muscarinic receptors may modulate not only cardiac vagal nerve activity but also sympathetic nerve activity in the skeletal muscle vasculature. PMID- 10821130 TI - Effect of combination therapy of angiotensin-converting enzyme inhibitor plus calcium channel blocker on urinary albumin excretion in hypertensive microalbuminuric patients with type II diabetes. AB - It has been demonstrated that antihypertensive treatment of hypertensive diabetic patients is quite effective in preventing macrovascular and microvascular complications and improving prognosis. Nevertheless, the target blood pressure level of antihypertensive treatment in hypertensive diabetic patients with microalbuminuria (i.e., with early diabetic nephropathy) remains to be established. In this study, we evaluated the effect of intensive blood pressure control (diastolic blood pressure <80 mmHg) on urinary albumin excretion in hypertensive, type II diabetic patients with microalbuminuria. We examined the effects of a combination therapy using an angiotensin-converting enzyme (ACE) inhibitor plus a long-acting calcium channel blocker (amlodipine), and compared them with the effect of an ACE inhibitor alone. Thirty hypertensive, type II diabetic patients with microalbuminuria were treated with either an ACE inhibitor alone (group I, n=17) or an ACE inhibitor plus amlodipine (group II, n=13) for 32 weeks. With treatment, blood pressures in both groups were significantly reduced, and diastolic blood pressure was lowered to a much greater extent in group II (76 +/- 2 mmHg) than in group I (83 +/- 2 mmHg, p < 0.05). Although the urinary albumin excretion rate was decreased in both groups, the decrease attained statistical significance only in group II (from 141 +/- 25 mg/day to 69 +/- 18 mg/day, p < 0.05); the extent of reduction in microalbuminuria during antihypertensive treatment was significantly greater in group II (50 +/- 10%) than in group I (14 +/- 13%, p < 0.05). In conclusion, this study showed that in hypertensive microalbuminuric type II diabetic patients, the combination of an ACE inhibitor plus amlodipine resulted in a more pronounced decreased in blood pressure (diastolic blood pressure <80 mmHg) and a greater reduction in urinary albumin excretion than did use of an ACE inhibitor alone. This combination strategy should thus be a more effective tool for obtaining optimal blood pressure control in patients with diabetic nephropathy. PMID- 10821131 TI - Participation in school sports clubs and related effects on cardiovascular risk factors in young males. AB - The effects of belonging to sports clubs on male high school students was evaluated. The relationships between the type and extent of school-based exercise were examined in conjunction with percent body fat, blood pressure (BP), and other key metabolic parameters. A total of 264 male Japanese high school students (age range: 17-18 years old) were studied. Percent body fat was measured and blood was collected in the fasting state during a routine health check. Subjects were divided into two groups. The exercise (E) group (n=150) included students who had belonged to a sports club during the past 2 years. The non-exercise (NE) group (n=114) included students who did not belong to a sports club during the past 2 years. The body mass index was significantly greater in group E (21.7 +/- 2.3 (SD) kg/m2) than in group NE (20.7 +/- 2.6 kg/m2, p < 0.01). However, the percent body fat in group E (13.6 +/- 3.4%) was significantly lower than that in group NE (14.9 +/- 3.8%, p < 0.01). The diastolic BP and heart rate in group E (64 +/- 7 mmHg, 70 +/- 11/min) were significantly lower in group E than in group NE (66 +/- 8 mmHg, p < 0.05; 76 +/- 14/min, p < 0.01). The serum triglyceride level was significantly lower, and the HDL cholesterol level was higher in group E than in group NE. The homeostasis model assessment (HOMA) index, used as an index of insulin resistance, was similar in the two groups. However, the level of the HOMA index was significantly lower among the 62 subjects in group E who preferred highly dynamic exercise (1.50 +/- 0.46) than it was among those in group NE (1.66 +/- 0.49, p < 0.05). Results indicate that belonging to sports clubs influences the BP and lipid profiles of adolescent males, as well as their percent body fat. In view of the reduction of cardiovascular risk factors, it is recommended that even young males practice regular exercise, especially aerobic exercise. PMID- 10821132 TI - Association of coronary risk factors and endothelium-dependent flow-mediated dilatation of the brachial artery. AB - Impaired endothelial function has been reported to be the initial step in atherosclerosis. Some coronary risk factors independently relate to impaired endothelial function. However, few studies have examined the association between coronary risk factors and endothelial function in patients who have multiple risk factors without clinical atherosclerosis. This study was undertaken to elucidate the relationship between accumulation of coronary risk factors and vascular endothelial dysfunction. We examined 101 subjects with one or more coronary risk factors 56.8 +/- 1.0 years old and 40 age-matched control subjects without coronary risk factors. We measured brachial artery diameter non-invasively using a 7.5-MHz ultrasound machine at rest, during reactive hyperemia caused by endothelium-dependent vasodilatation, and after sublingual administration of nitroglycerin, which causes endothelium-independent vasodilatation. The percentage change in flow-mediated diameter (%FMD; deltaD/D x 100), in subjects with one or more coronary risk factors was significantly lower than that in control subjects(4.8 +/- 0.3% vs. 6.7 +/- 0.5%, p < 0.01). Endothelium independent vasodilatation by nitroglycerin did not differ between the two groups. Endothelial function was impaired according to the accumulation of coronary risk factors. On multiple regression analysis, the number of risk factors, age, and brachial artery diameter at rest showed significant correlation with %FMD. Our results suggest that an accumulation of coronary risk factors was significantly related to impairment of endothelial function. PMID- 10821133 TI - Effects of coronary blood flow on left ventricular function in essential hypertensive patients. AB - To elucidate the mechanism of left ventricular dysfunction associated with left ventricular hypertrophy in hypertension, coronary blood flow (CBF) and left ventricular mass (LVM) were measured in 62 patients with essential hypertension (mean age, 54 +/- 13 years) and 22 normotensive control subjects (mean age, 57 +/ 13 years). According to the indicator fractionation principle, CBF/cardiac output (CO), estimated on the basis of the ratio of myocardial uptake/total injected dose of thallium-201 (% cardiac uptake), was measured. CBF and CBF per 100 g of myocardium (unit CBF) were calculated according to the following formulas: CBF=% cardiac uptake x CO, and unit CBF=(CBF/LVM) x 100, where CO and LVM are echocardiographically determined. Midwall fractional shortening (FS) and isovolumic relaxation time (IRT) were calculated as the indices of systolic and diastolic functions. CBF was greater in hypertensives than in controls (218.2 +/- 74.0 vs. 187.4 +/- 40.4 ml/min, p < 0.05), though unit CBF was smaller in hypertensives than in controls (99.1 +/- 22.0 vs. 141.2 +/- 31.6 ml/min/100 g, p < 0.0001). Multiple regression analyses showed that unit CBF was the most potent predictor of both midwall FS and IRT. A positive correlation was found between midwall FS and unit CBF (r=0.669, p < 0.0001), and a negative correlation between IRT and unit CBF (r=-0.579, p < 0.0001). In conclusion, myocardial ischemia reflected by the decrease in unit CBF may be closely related to left ventricular dysfunction in patients with essential hypertension. PMID- 10821134 TI - Atypical aortic coarctation with resistant hypertension treated with axilloiliac artery bypass. AB - A 68-year-old woman was found to have atypical coarctation of the aorta, accompanied by systolic hypertension of the upper extremities despite administration of five types of antihypertensive drugs. Since the systolic hypertension was resistant to the conventional antihypertensive therapy, axilloiliac artery bypass grafting with a subcutaneous tunnel was performed to alleviate the pressure gradient. Systolic blood pressure was successfully reduced and hypertension was controlled after surgery. PMID- 10821135 TI - Cellular mechanisms of cardioprotection afforded by inhibitors of angiotensin converting enzyme in ischemic hearts: role of bradykinin and nitric oxide. AB - Angiotensin converting enzyme (ACE) inhibitors inhibit the degradation of bradykinin and contribute to accumulation of bradykinin and NO, both of which may be beneficial for diseased hearts. To test this idea, we administered imidaprilat and cilazaprilat, respectively to the canine ischemic myocardium. In the open chest dogs with low constant coronary perfusion pressure (CPP, from 104 +/- 3 to 42 +/- 3 mmHg), coronary blood flow (CBF, 91 +/- 1 to 32 +/- 2 ml/100 g/min), fractional shortening (FS), and lactate extraction ratio (LER) decreased. Either imidaprilat or cilazaprilat increased CBF, FS, and LER with increases in cardiac bradykinin and NO levels. The beneficial effects of ACE inhibitors were blunted by either L-NAME (an inhibitor of NO synthase) and HOE140 (an inhibitor of bradykinin receptors), respectively. ACE inhibitors, on the other hand, are reported to attenuate the severity of myocardial stunning, which effect is partially attributable to bradykinin- and NO-dependent mechanisms. Further, ACE inhibitors limited infarct size following coronary occlusion and reperfusion. This infarct size-limitation was blunted by either L-NAME and IBTX (the antagonist of K(Ca) channels). Bradykinin is also reported to close K(Ca) channels. Thus, we concluded that ACE inhibitors attenuate both reversible and irreversible myocardial cellular injury via bradykinin/NO-dependent mechanisms. In experimental and clinical settings, the cardioprotective effects of ACE inhibitors on the diseased heart may be attributable to these mechanisms. PMID- 10821136 TI - No correlation of polymorphism of angiotensin-converting enzyme genes with left ventricular hypertrophy in essential hypertension. AB - To investigate the correlation of polymorphism of angiotensin-converting enzyme (ACE) genes with left ventricular hypertrophy in essential hypertension, 151 patients with essential hypertension were studied. ACE genotypes were determined by PCR technology and diastolic left ventricular diameter (DLVd), systolic left ventricular diameter (SLVd), interseptal ventricular thickness (IVS), and left ventricular posterior wall thickness (LVPW) were scanned by echocardiography. Left ventricular mass (LVM) and the left ventricular mass index (LVMI) were calculated from echocardiographic findings. Results revealed that DLVd, SLVd, IVS, LVPW, LVM, and LVMI of the DD genotype group were 49.9 +/- 5.6 mm, 30.5 +/- 6.5 mm, 11.2 +/- 1.6 mm, 11.7 +/- 1.5 mm, 259.5 +/- 62.1 g, 92.7 +/- 23.5 g/m2, respectively. DLVd, SLVd, IVS, LVPW, LVM, and LVMI of the ID genotype group were 8.9 +/- 5.3 mm, 31.5 +/- 5.2 mm, 11.4 +/- 1.7 mm, 11.9 +/- 1.6 mm, 261.3 +/- 70.3 g, and 94.9 +/- 25.8 g/m2, respectively, and DLVd, SLVd, IVS, LVPW, LVM, and LVMI of the II genotype group are 48.9 +/- 5.5 mm, 31.8 +/- 6.5 mm, 11.1 +/- 1.9 mm, 11.5 +/- 1.8 mm, 250.8 +/- 82.5 g and 90.8 +/- 30.1 g/m2 respectively. There was no significant difference between the ID, DD and II genotype groups as regards DLVd, SLVd, IVS, LVPW, LVM, and LVMI (p > 0.05). These findings indicate that there is no association between the ACE gene and left ventricular hypertrophy in essential hypertension occurring in the Chinese population. PMID- 10821137 TI - Correlative factors of insulin resistance in essential hypertension. AB - Essential Hypertension (EH) is correlated with a metabolic disturbance characterized by insulin resistance (IR). In this study, there were observed in 47 subjects with EH and 30 subjects with normal blood pressure. Serum levels of insulin-like growth factor-1 (IGF-1), serum levels of growth hormone (GH), the activity of erythrocyte insulin receptors (EIR), and ATP levels in erythrocytes, the insulin sensitivity index (ISI) was used to study the correlative factors of essential hypertension. 1. Among patients with EH, ISI, GH, and low-affinity insulin binding sites of EIRs (RT2) were found to be in significantly lower amounts, IGF-1 levels and the KD2 of the erythrocyte insulin receptors were noted to be significantly higher. Compared with the control group, there was a marked difference between EH group and the control group. However, no statistical difference was observed between the hypertensive group and the group with normal blood pressure as regards erythrocyte ATP levels, high-affinity insulin binding sites of EIRs (RT1), and the KD1 of EIRs. 2. In the hypertensive group, the ISI was negatively correlated with mean arterial blood pressure (MBP), a family history of hypertension, the body mass index (BMI), the waist-hip ratio (WHR) and IGF-1 levels (r=-0.614delta, -0.354**, -0.386**, -0.472**, -0.298*, delta p < 0.001, **p < 0.01, *p < 0.05), were positively correlated with RT2 and GH levels (r=0.301**, 0.275*, **p < 0.01, *p < 0.05). There were no statistically significant differences between ISI and age, sex, smoking history, drinking, RT1, KD1, and ATP levels in erythrocytes. 3. The ISI was used as the dependent variable in multiple linear stepwise regression analysis. MBP (X1), a family history of EH (X2), WHR (X3), GH (X4), IGF-1 (X5), RT2 (X6), and the body mass index (X7) was used as independent variables. X1, X2, X3, X5, X6, and X7 were used in the equations. The results indicate that patients with EH also tend to have IR. We suggest that MBP, a family history of hypertension, BMI, WHR, IGF-1, and RT2 might be independent factors affecting IR in cases of essential hypertension. PMID- 10821138 TI - Association of polymorphism in the promoter region of the apolipoprotein E gene with diastolic blood pressure in normotensive Japanese. AB - The epsilon4 allele of apolipoprotein E (APOE) is reported to be a genetic risk factor of atherosclerosis through hyperlipidemia and late-onset Alzheimer's dementia. A recent report showed that a genetic variant (A -491T) in the promoter region of the APOE gene increases the risk of Alzheimer's disease. In the present study, we examined whether these APOE polymorphisms were genetically involved in essential hypertension. Japanese hypertensives (n=180) with a family history of hypertension and normotensive controls (n=195, sex and age matched with hypertensives) were recruited from the outpatients of Osaka University Hospital, and an informed consent to participate in the study was obtained from each person. APOE polymorphisms were determined using polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). The frequencies of the A 491 allele in hypertensives and normotensives were 0.98 and 0.97, respectively, and the TT/-491 genotype was not found in either group. No significant differences between hypertensives and normotensives were observed in allele frequencies in either APOE polymorphism; however, the mean diastolic blood pressure in normotensive subjects with AA/-491 was significantly higher than in the subjects with AT/-491 (p < 0.01). These results suggest that the presence of the APOE promoter polymorphism is not a major risk factor for hypertension but that it does have some minor effect on basal blood pressure variation. PMID- 10821139 TI - Oral taurine supplementation prevents the development of ethanol-induced hypertension in rats. AB - Taurine is known to lower blood pressure in essential hypertension and some experimental hypertensive models. Taurine has also been reported to activate aldehyde dehydrogenase and to inhibit the elevation of plasma acetaldehyde concentration after ethanol intake. Because acetaldehyde, the first metabolite of ethanol, is suspected to be responsible for many adverse effects of alcohol consumption, we examined the effect of taurine supplementation on ethanol-induced hypertension and abnormalities in the intracellular cation metabolism in Witar Kyoto rats. In Study 1, systolic blood pressure and intraplatelet free calcium were significantly higher in rats who received 15% ethanol in drinking water than in control rats. Oral taurine supplementation (1% taurine and 15% ethanol in drinking water) completely prevented the development of ethanol-induced hypertension. Intraerythrocyte sodium and intraplatelet free calcium were significantly decreased in taurine-supplemented rats as compared with rats who received 15% ethanol only. In Study 2, hemoglobin-associated acetaldehyde (HbAA) was measured as a marker of protein-bound acetaldehyde. HbAA was significantly elevated in rats who received 5% ethanol in drinking water as compared with control rats. Taurine supplementation (1% taurine and 5% ethanol in drinking water) significantly decreased HbAA. Our findings suggest that the oral supplementation of taurine prevents ethanol-induced hypertension by decreasing protein bound acetaldehyde and altering the cation handling by the membrane. PMID- 10821141 TI - Oral contraceptives and prevention of implantation. PMID- 10821140 TI - Assessment of in vivo oxidative stress in hypertensive rats and hypertensive subjects in Tanzania, Africa. AB - Oxidative stress has been reported to be involved in not only cardiovascular diseases but in hypertension, which is a major risk for cardiovascular diseases. Urinary 8-hydroxy-2'-deoxyguanosine (8-OHdG) has been recognized as a sensitive biomarker of oxidative DNA damage and also of oxidative stress. In the present study, we assessed the oxidative stress in human subjects with hypertension and in hypertensive rats. In stroke-prone spontaneously hypertensive rats at the age of 14 weeks, the excretion of urinary 8-OHdG was significantly (p < 0.05) increased compared with that in age-matched normotensive Wistar-Kyoto rats. Next, we investigated the relationship between oxidative DNA damage and cardiovascular risk factors among Tanzanians aged 46-58 years in a population study carried out in 1998 in at Dar es Salaam, Tanzania, according to the WHO-CARDIAC Study Protocol. Sixty subjects (male/female, 28/32) were selected by SPSS Base 8.0 from those who completed a 24-h urine collection. The 24-h urinary 8-OHdG of the hypertensive subjects (SBP > or =140 mmHg and/or DBP > or =90 mmHg) was significantly (p < 0.05) higher than that of the normotensive subjects (SBP <140 mmHg and DBP <90 mmHg) after adjusting for age and gender (Hypertensives: 17.31 +/- 2.0 ng/mg creatinine, n=38; Normotensives: 10.10 +/- 2.64 ng/mg creatinine, n=22). Oxidative stress was thought to be involved in hypertensive subjects and in hypertensive rats. PMID- 10821142 TI - Oral contraceptives and prevention of implantation. PMID- 10821143 TI - An uninvited guest in the ear. PMID- 10821144 TI - Asymptomatic microscopic hematuria. PMID- 10821145 TI - Asymptomatic microscopic hematuria. PMID- 10821146 TI - Factors at play in the athletic preparticipation examination. PMID- 10821147 TI - It won't be me next time: an opinion on preparticipation sports physicals. PMID- 10821148 TI - Family practice and the elimination of tuberculosis. PMID- 10821149 TI - Identification and management of tuberculosis. AB - Although the resurgence of tuberculosis in the early 1990s has largely been controlled, the risk of contracting this disease remains high in homeless persons, recent immigrants and persons infected with the human immunodeficiency virus (HIV). Purified protein derivative testing should be targeted at these groups and at persons with known or suspected exposure to active tuberculosis. Most patients with latent tuberculosis are treated with isoniazid administered daily for nine months. In patients with active tuberculosis, the initial regimen should include four drugs for at least two months, with subsequent therapy determined by mycobacterial sensitivities and clinical response. To avoid harmful drug interactions, regimens that do not contain rifampin may be employed in HIV infected patients who are taking protease inhibitors or nonnucleoside reverse transcriptase inhibitors. To maximize compliance and minimize the emergence of mycobacterial drug resistance, family physicians should consider using directly observed therapy in all patients with tuberculosis. PMID- 10821150 TI - The preparticipation athletic evaluation. AB - A comprehensive medical history that includes questions about a personal and family history of cardiovascular disease is the most important initial component of the preparticipation athletic evaluation. Additional questions should focus on any history of neurologic or musculoskeletal problems. A limited physical examination should emphasize cardiac auscultation with provocative maneuvers to screen for hypertrophic cardiomyopathy. This condition is the most common cause of sudden death in young male athletes. Other components of the physical examination include an evaluation of the spine and extremities. Screening tests such as electrocardiography, treadmill stress testing and urinalysis are not indicated in the absence of symptoms or a significant history of risk factors. Specific conditions that would exclude or limit athletic participation include hypertrophic cardiomyopathy, long QT interval syndrome, concussion, significant knee injury, sickle cell disease and uncontrolled seizures. Overall, about 1 percent of athletes who are screened are completely disqualified from sports participation. PMID- 10821151 TI - Treatment of seborrheic dermatitis. AB - Seborrheic dermatitis is a chronic inflammatory disorder affecting areas of the head and trunk where sebaceous glands are most prominent. Lipophilic yeasts of the Malassezia genus, as well as genetic, environmental and general health factors, contribute to this disorder. Scalp seborrhea varies from mild dandruff to dense, diffuse, adherent scale. Facial and trunk seborrhea is characterized by powdery or greasy scale in skin folds and along hair margins. Treatment options include application of selenium sulfide, pyrithione zinc or ketoconazole containing shampoos, topical ketoconazole cream or terbinafine solution, topical sodium sulfacetamide and topical corticosteroids. PMID- 10821152 TI - Practical use of the pessary. AB - The pessary is an effective tool in the management of a number of gynecologic problems. The pessary is most commonly used in the management of pelvic support defects such as cystocele and rectocele. Pessaries can also be used in the treatment of stress urinary incontinence. The wide variety of pessary styles may cause confusion for physicians during the initial selection of the pessary. However, an understanding of the different styles and their uses will enable physicians to make an appropriate choice. Complications can be minimized with simple vaginal hygiene and regular follow-up visits. PMID- 10821153 TI - Bisphosphonates: safety and efficacy in the treatment and prevention of osteoporosis. AB - Osteoporosis affects more than 28 million Americans. With the advent of accessible and affordable diagnostic studies, awareness and recognition of this disease by patients and clinicians are growing. Osteoporotic fractures of the spine and hip are costly and associated with significant morbidity and mortality. Over the past decade, a surge of new antiosteoporotic drugs have been labeled or are awaiting labeling by the U.S. Food and Drug Administration. One class of agents used to treat osteoporosis is the bisphosphonates, which inhibit bone resorption, cause an increase in bone mineral density and reduce the risk of future fractures caused by aging, estrogen deficiency and corticosteroid use. Overall, bisphosphonates have been shown to have a strong safety and tolerability profile. PMID- 10821154 TI - New classification and update on the quinolone antibiotics. AB - The newer fluoroquinolones have broad-spectrum bactericidal activity, excellent oral bioavailability, good tissue penetration and favorable safety and tolerability profiles. A new four-generation classification of the quinolone drugs takes into account the expanded antimicrobial spectrum of the more recently introduced fluoroquinolones and their clinical indications. First-generation drugs (e.g., nalidixic acid) achieve minimal serum levels. Second-generation quinolones (e.g., ciprofloxacin) have increased gram-negative and systemic activity. Third-generation drugs (e.g., levofloxacin) have expanded activity against gram-positive bacteria and atypical pathogens. Fourth-generation quinolone drugs (currently only trovafloxacin) add significant activity against anaerobes. The quinolones can be differentiated within classes based on their pharmacokinetic properties. The new classification can help family physicians prescribe these drugs appropriately. PMID- 10821155 TI - Noise-induced hearing loss. AB - Hearing loss caused by exposure to recreational and occupational noise results in devastating disability that is virtually 100 percent preventable. Noise-induced hearing loss is the second most common form of sensorineural hearing deficit, after presbycusis (age-related hearing loss). Shearing forces caused by any sound have an impact on the stereocilia of the hair cells of the basilar membrane of the cochlea; when excessive, these forces can cause cell death. Avoiding noise exposure stops further progression of the damage. Noise-induced hearing loss can be prevented by avoiding excessive noise and using hearing protection such as earplugs and earmuffs. Patients who have been exposed to excessive noise should be screened. When hearing loss is suspected, a thorough history, physical examination and audiometry should be performed. If these examinations disclose evidence of hearing loss, referral for full audiologic evaluation is recommended. PMID- 10821156 TI - An approach to drug abuse, intoxication and withdrawal. AB - The symptomatic effects of drug abuse are a result of alterations in the functioning of the following neurotransmitters or their receptors: acetylcholine, dopamine, gamma-aminobutyric acid, norepinephrine, opioids and serotonin. Anticholinergic drugs antagonize acetylcholine receptors. Dissociative drugs affect all transmitter sites. Opiates act on both opioid and adrenergic receptor sites. Psychedelic drugs stimulate serotonin release, and sedative-hypnotic drugs potentiate the gamma-aminobutyric acid receptor. Specific signs and symptoms are associated with the neurotransmitters and receptors affected by each drug class. By recognizing symptomatic changes related to particular neurotransmitters and their receptors, family physicians can accurately determine the drug class and intervene appropriately to counteract drug-induced effects. PMID- 10821157 TI - Medical surveillance in work-site safety and health programs. AB - Businesses frequently call on family physicians to provide employee health services at the work site or in the clinician's office. These services include medical screening (detection of dysfunction or disease before an employee would ordinarily seek medical care) and medical surveillance (analysis of health information to identify workplace problems that require targeted prevention). Such services can transform acute care and routine screening activities into opportunities for primary prevention when they are integrated into the broader framework of work-site safety and health programs. Components of these programs include management commitment, employee participation, hazard identification and control, employee training and program evaluation. For optimal program success, family physicians must communicate with frontline safety officers and have first hand knowledge of the workplace and its hazards. Professional and technical resources are available to guide the family physician in the role of medical surveillance program coordinator. PMID- 10821158 TI - Bilious vomiting in the newborn: rapid diagnosis of intestinal obstruction. AB - Bilious vomiting in newborns is an urgent condition that requires the immediate involvement of a team of pediatric surgeons and neonatologists for perioperative management. However, initial detection, evaluation and treatment are often performed by nurses, family physicians and general pediatricians. Bilious vomiting, with or without abdominal distention, is an initial sign of intestinal obstruction in newborns. A naso- or orogastric tube should be placed immediately to decompress the stomach. Physical examination should be followed by plain abdominal films. Dilated bowel loops and air-fluid levels suggest surgical obstruction. Contrast radiography may be required. Duodenal atresia, midgut malrotation and volvulus, jejunoileal atresia, meconium ileus and necrotizing enterocolitis are the most common causes of neonatal intestinal obstruction. PMID- 10821159 TI - Postpartum axillary masses. PMID- 10821160 TI - ACC/AHA revise guidelines for coronary bypass surgery. American College of Cardiology/American Heart Association. PMID- 10821161 TI - What your patients don't tell you. PMID- 10821162 TI - Adult immunization--influenza vaccine. PMID- 10821163 TI - Assessment of specificity of eight chemical inhibitors using cDNA-expressed cytochromes P450. AB - 1. The selectivity of eight chemical inhibitors has been extensively evaluated with 10 cDNA-expressed human cytochrome P450 isoforms (CYP). The results indicate that sulphaphenazole, quinidine and alpha-naphthoflavone are selective inhibitors of CYP2C9 (IC50 = 0.5-0.7 microM), CYP2D6 (0.3-0.4 microM) and CYP1A (0.05-5 microM) respectively on the basis of the IC50, which are much lower than those of other P450 isoforms (> 10-fold). 2. Ketoconazole exhibited potent inhibition of both CYP3A4-catalysed metabolism of phenanthrene, testosterone, diazepam (IC50 = 0.03-0.5 microM) and CYP1A1-catalysed deethylation of 7-ethoxycoumarin (0.33 microM). The selectivity of ketoconazole for other P450s was highly related to the concentration used. 3. Diethyldithiocarbamate, orphenadrine and furafylline were shown separately to be less selective inhibitors of CYP2E1, CYP2B6 and CYP1A isoforms by a broad range of IC50 that overlap those observed with other P450 isoforms. 4. Furafylline, quinidine and alpha-naphthoflavone activated CYP3A4 catalysed phenanthrene metabolism by 1.7-, 2- and 15-fold respectively. 5. The selectivity of orphenadrine and ketoconazole was further examined by using inhibitory monoclonal antibodies (MAb). Inhibitory MAb specific for the individual P450 isoforms may be of greater value than chemical inhibitors. PMID- 10821164 TI - Thyroid hormone modulation of rat sulphotransferase mRNA expression. AB - 1. Thyroid hormones modulate sulphotransferase (SULT) enzyme expression. Specific substrates are not available for the study of the SULT isoforms, so the regulation of hepatic SULT mRNA expression by thyroid hormones was examined by Northern blot analysis with oligonucleotide probes specific for each SULT mRNA, including male-dominant phenol SULT (1A1, 1C1, 1E2), female-dominant hydroxysteroid SULT (20/21, 40/41, 60), and a non-sex-dependent SULT, 1B1. The male and female rat were either untreated, thyroidectomized (TX), or TX and given thyroid hormones (thyroxine [T4, 20 microg kg(-1) day(-1)] and 3,5,3' triiodothyronine [T3, 5 microg kg(-1) day(-1)]). 2. With regard to phenol SULTs, expression of SULT1A1 or SULT1B1 mRNA was not altered in either sex by TX. TX increased SULT1E2 mRNA expression 3-fold in the male and 2.5-fold in the female rat. The increase in SULT1E2 mRNA was partially reversed by infusion of T3/T4 in the male, and was not reversed in the female. 3. With regard to hydroxysteroid SULTs, TX decreased expression of SULT20/21 mRNA in the male rat by 70 and 60% in the female, and these decreases were reversed by T3/T4 infusion. SULT40/41 mRNA expression increased in the male rat 3-fold and decreased in the female TX rat by 25%. SULT60 mRNA expression increased 3-fold by TX in the female rats. The effects of TX on SULT40/41 and SULT60 mRNA expression were reversed by infusion of T2/T4. 5. Thus, phenol sulphotransferases were not markedly affected by thyroid hormones except for SULT1E2, but each hydroxysteroid sulphotransferase isoform was affected by thyroidectomy. Therefore, thyroid hormones regulate SULT gene expression in an isoform-specific manner. PMID- 10821165 TI - Reduction of stilbene oxide and styrene oxide to the corresponding alkenes by intestinal bacteria. AB - 1. This study provides the first evidence that stilbene oxide and styrene oxide are reductively metabolized to the corresponding alkenes by intestinal bacteria in animals. 2. When trans- or cis-stilbene oxide were incubated with the caecal contents of rat under anaerobic conditions, both trans- and cis-stilbene were isolated from the incubation mixture. Styrene oxide was also reduced to styrene by rat caecal contents. 3. Caecal contents of mouse, hamster and guinea pig also exhibited alkene oxide reductase activities toward cis- and trans-stilbene oxides, and styrene oxide. In contrast, liver microsomes or cytosol exhibited no epoxide reductase activities toward these substrates. 4. Seven pure strains of intestinal bacteria exhibited alkene oxide reductase activities of varying degrees under anaerobic conditions, with the highest activity being observed in Clostridium sporogenes. 5. Cell-free extracts of either the intestinal bacteria in rat caecal contents or C. sporogenes exhibited reductase activity when supplemented with both NAD(P)H and FMN under anaerobic conditions. Reductase activity was also observed on addition of the photochemically reduced form of FMN instead of both NAD(P)H and FMN. PMID- 10821166 TI - Metabolism of roquinimex in mouse and rat: an in vitro/in vivo comparison. AB - 1. In vitro studies with roquinimex, an immuno-modulator, in liver microsomes from mouse and rat were conducted to evaluate the primary metabolism and compare the metabolite pattern as well as the rate of metabolism with the in vivo pharmacokinetics of the compound in these two species. 2. In the presence of NADPH, roquinimex was metabolized to six primary metabolites (R1-6) by liver microsomes from mouse and rat. The formation of these metabolites was qualitatively similar in both species, and was greatly enhanced by pretreatment with PCN, an inducer of cytochrome P4503A. 3. The identification of the R1-6 demonstrated that roquinimex had been hydroxylated and demethylated. Hydroxylation at different sites of the quinoline moiety was the dominating reaction in both species. 4. Comparison of the resulting microsomal intrinsic clearance of 0.3 micromol mg(-1) protein min(-1) in mouse liver microsomes, versus 0.03 micromol mg(-1) protein min(-1) in rat liver microsomes demonstrated that the mouse possesses about a 10-fold greater metabolic capacity for roquinimex than the rat. 5. The in vivo pharmacokinetics of roquinimex demonstrated a 7-fold higher clearance in mouse than in the rat (82 ml h(-1) kg( 1) in mouse, 10.6 ml h(-1) kg(-1) in rat), which is in concordance with the in vitro findings. PMID- 10821167 TI - Enantioselective liver microsomal sulphoxidation of albendazole in cattle: effect of nutritional status. AB - 1. The enantioselective liver microsomal sulphoxidation of the benzimidazole anthelmintic, albendazole (ABZ), by cattle liver microsomes has been investigated. The influence of nutritional condition on this biotransformation process was also characterized. 2. ABZ was oxidized to its sulphoxide metabolite (ABZSO) in a NADPH concentration-dependent reaction and the (+) and (-) ABZSO enantiomers formed were identified. 3. Vmax (0.27 nmol ABZSO formed per min x mg( 1) microsomal protein) and Km (15.10 microM) for ABZ sulphoxidation by cattle liver microsomes were obtained. Different Vmax (0.11 and 0.16 nmol x min(-1) x mg(-1)) and Km (9.40 and 26.70 microM) characterized the enantioselective formation of (+) and (-) ABZSO antipodes, respectively. 4. Free fatty acid (FFA) concentrations and beta-hydroxybutyrate concentrations (beta-OHB) in serum and liver homogenates were significantly higher in feed-restricted (poor nutritional condition) compared with control animals in an optimal nutritional status. Serum protein concentrations and liver cytosolic glucose 6-phosphate dehydrogenase (G6PD) activity were significantly lower in the feed-restricted compared with control calf. 5. Animal nutritional condition affected the pattern of ABZ sulphoxidation. A higher Km for (total) ABZSO and (+) ABZSO production was observed in the calf subjected to a period of undernutrition. 6. A nutritionally induced impairment in the affinity of microsomal mixed-function oxidases responsible of ABZ oxidation may be responsible for the observed changes in the liver microsomal sulphoxidation of ABZ in the feed-restricted calf. Furthermore, undernutrition may affect primarily the FMO-mediated formation of (+) ABZSO. These in vitro observations agree with the changes observed in vivo following the administration of ABZ to the calf subjected to a dietary restriction. PMID- 10821168 TI - Metabolism of Cyanox in rat. II. Sex-related differences in oxidative dearylation and desulphuration. AB - 1. To examine the metabolites of Cyanox (O-4-cyanophenyl O,O-dimethyl phosphorothioate, cyanophos, CYAP) in brain, liver, blood cells and plasma during the early toxic period, the male and female rat was administered a single oral dose of [phenyl-14C]Cyanox at dose levels of 50 mg/kg and killed 5, 10 and 20 min thereafter. 2. Sex-related differences in the concentrations of metabolites were observed. Cyanoxon, produced by oxidative desulphuration, was observed in the brains of both sexes at all time points, but the concentrations were 2-6 times higher in the male. The same metabolite was detected in the liver, blood cells and plasma of the male but not the female. The total concentrations of oxidative dearylation metabolites (4-cyanophenol + 4-cyanophenylsulphate + glucuronide of 4 cyanophenol) in plasma, blood cell, brain and liver were larger in the male at all time points than those in the female, whereas the reverse was the case for demethylated metabolites (desmethylcyanox + desmethylcyanoxon) in all tissues except for the brain. 3. Studies of the in vitro metabolism of Cyanox revealed no sex-related difference for hepatic cytosolic fractions in terms of the major in vitro metabolic reaction, demethylation. On the other hand, the major reactions in microsomal fractions, oxidative desulphuration and oxidative dearylation, were significantly (2-3 times) greater in the male than in the female. 4. Oxidative desulphuration and oxidative dearylation, involving cytochrome P450 enzymes, were inhibited by male-specific rat CYP2C11 antiserum. The degree of inhibition was more pronounced in the male case. Thus, the results strongly suggest that the 2C family of cytochrome P450 (male, CYP2C11 and CYP2C13; female, CYP2C12) contributes to oxidative desulphuration and dearylation of cyanox in the rat and that the activity of male-specific CYP2C11 (and CYP2C13) is greater than that of female-specific CYP2C12. The consequent greater formation of cyanoxon in the male is consistent with the higher toxicity in this sex. PMID- 10821169 TI - Urinary metabolites of a novel quinoxaline non-nucleoside reverse transcriptase inhibitor in rabbit, mouse and human: identification of fluorine NIH shift metabolites using NMR and tandem MS. AB - 1. The urinary metabolites of (S)-2-ethyl-7-fluoro-3-oxo-3,4-dihydro-2H quinoxaline-carboxylic acid isopropylester (GW420867X) have been investigated in samples obtained following oral administration to rabbit, mouse and human. GW420867X underwent extensive biotransformation to form hydroxylated metabolites and glucuronide conjugates on the aromatic ring, and on the ethyl and isopropyl side-chains in all species. In rabbit urine, a minor metabolite was detected and characterized as a cysteine adduct that was not observed in mouse or man. 2. The hydroxylated metabolites and corresponding glucuronide conjugates were isolated by semi-preparative HPLC and characterized using NMR, LC-NMR and LC-MS/MS. The relative proportions of fluorine-containing metabolites were determined in animal species by 19F-NMR signal integration. 3. The fluorine atom of the aromatic ring underwent NIH shift rearrangement in the metabolites isolated and characterized in rabbit, mouse and human urine. 4. The characterization of the NIH shift metabolites in urine enabled the detection and confirmation of the presence of these metabolites in human plasma. PMID- 10821170 TI - Identification of turkey biliary metabolites of ractopamine hydrochloride and the metabolism and distribution of synthetic [14C]ractopamine glucuronides in the turkey. AB - 1. The bile duct cannulated turkey poult (n = 3) dosed orally with [14C]ractopamine HCl [(1R*,3R*),(1R*,3S*)-4-hydroxy-alpha-[[[3-(4 hydroxy[14C]phenyl)-1-methy lpropyl]amino]methyl]-benzenemethanol hydrochloride; 19.9 mg; 9.28 microCi] excreted 37.4 +/- 12.1% (mean +/- SD) of the administered radioactivity in bile by 24 h post-dosing. 2. A mono-glucuronide, conjugated at C 10 (the methylpropylamino phenol) of ractopamine, accounted for 76.6% of biliary radioactivity. 3. Urine collected from the colostomized turkey poult (n = 3) orally dosed with synthetic [14C]ractopamine-glucuronides (10.1 mg; 3.6 microCi) contained 11.9 +/- 1.0% (mean +/- SD) of the administered radioactivity 24 h after dosing, indicating that some absorption of radioactivity occurred. Faeces contained 60.6% of the administered radioactivity and carcasses (with gastrointestinal tracts) contained 23.3% of the starting radioactivity. 4. Five colostomized poults were fitted with bile duct cannulas and were dosed intraduodenally with 10.2 mg (3.6 microCi) synthetic [14C]ractopamine glucuronides. Urine and bile contained 15.5 +/- 2.2 and 16.8 +/- 2.1% respectively of the administered radiocarbon by 24 h post-dosing. Faeces contained 54.3% of the administered radioactivity. Total absorption of the dosed radioactivity averaged 33.4%. 5. Bile and urine collected from the colostomized, bile-duct cannulated bird contained mainly ractopamine glucuronides. Indirect evidence suggests that the dosed ractopamine glucuronides were not absorbed intact. PMID- 10821172 TI - Quantification of total genomic DNA and selected repetitive sequences reveals concurrent changes in different DNA families in indica and japonica rice. AB - This paper describes a fluorescence in situ hybridization (FISH) analysis of three different repetitive sequence families, which were mapped to mitotic metaphase chromosomes and extended DNA fibers (EDFs) of the two subspecies of rice (OrYza sativa), indica and japonica (2n = 2x = 24). The repeat families studied were (1) the tandem repeat sequence A (TrsA), a functionally non significant repeat; (2) the [TTTA-GGG]n telomere sequence, a non-transcribed, tandemly repeated but functionally significant repeat; and (3) the 5S ribosomal RNA (5S rDNA). FISH of the TrsA repeat to metaphase chromosomes of indica and japonica cultivars revealed clear signals at the distal ends of twelve and four chromosomes, respectively. As shown in a previous report, the 17S ribosomal RNA genes (17S rDNA) are located at the nucleolus organizers (NORs) on chromosomes 9 and 10 of the indica cultivar. However, the japonica rice lacked the rDNA signals on chromosome 10. The size of the 5S rDNA repeat block, which was mapped on the chromosome 11 of both cultivars, was 1.22 times larger in the indica than in the japonica genome. The telomeric repeat arrays at the distal ends of all chromosome arms were on average three times longer in the indica genome than in the japonica genome. Flow cytometric measurements revealed that the nuclear DNA content of indica rice is 9.7% higher than that of japonica rice. Our data suggest that different repetitive sequence families contribute significantly to the variation in genome size between indica and japonica rice, though to different extents. The increase or decrease in the copy number of several repetitive sequences examined here may indicate the existence of a directed change in genome size in rice. Possible reasons for this phenomenon of concurrent evolution of various repeat families are discussed. PMID- 10821171 TI - A spindle pole body-associated protein, SNAD, affects septation and conidiation in Aspergillus nidulans. AB - The nudA1 mutation in the cytoplasmic dynein heavy chain gene inhibits nuclear migration, colony growth and asexual sporulation (conidiation) in the filamentous fungus Aspergillus nidulans. It also alters the location of the first cell division event (septation) and prevents nucleation of tip cells. We showed previously that a suppressor of nudA1, snaD290, partially reversed the nuclear migration defect and partially restored colony growth. We have now demonstrated that the snaD290 mutation also delays septation and restores the septum to its normal position, allowing tip cells to be nucleated. Although snaD290 does not affect nuclear migration or vegetative hyphal growth, it almost completely inhibits conidiation. We propose that the SNAD protein participates in septation, and is essential for asexual spore formation. SnaD encodes a novel 76-kDa coiled coil protein (SNAD) that is located at the spindle pole body throughout the cell cycle. Therefore, our results suggest that proteins at the spindle pole body are likely to be involved in temporal regulation of septation in A. nidulans. PMID- 10821173 TI - A linear RNA replicon from the oomycete Phytophthora infestans. AB - An unusual RNA element was discovered in an isolate of the oomyceteous fungus Phytophthora infestans. The RNA exists predominantly as single-stranded molecules of about 625 nucleotides with complementary strands present at a ratio of approximately 130:1. Gel mobility and PCR assays indicated that the element was linear. The RNA appeared to be an autonomous element, since P. infestans DNA did not contain cross-hybridizing sequences. Standard methods for virus purification yielded no evidence for encapsidation of the RNA, or for other virus particles in the isolate bearing the replicon. The replicon contained polyU and polyA tracts at its 5' and 3' termini, respectively, with a central region that had a GC content of 47%, and lacked obvious ORFs. Two-thirds of the replicon co-purified with nuclei, at about 200 copies per nucleus, while one-third resided in a cytoplasmic but non-mitochondrial location. Maternal inheritance was observed in sexual crosses, with a few exceptions. The replicon was not widely distributed throughout the species and had little effect on growth or pathogenicity. The data suggest that the RNA is best characterized as a novel linear RNA plasmid. PMID- 10821174 TI - Tools for chloroplast transformation in Chlamydomonas: expression vectors and a new dominant selectable marker. AB - Reverse-genetic studies of chloroplast genes in the green alga Chlamydomonas reinhardtii have been hampered by the paucity of suitable selectable markers for chloroplast transformation. We have constructed a series of vectors for the targeted insertion and expression of foreign genes in the Chlamydomonas chloroplast genome. Using these vectors we have developed a novel selectable marker based on the bacterial gene aphA-6, which encodes an aminoglycoside phosphotransferase. The aphA-6 marker allows direct selection for transformants on medium containing either kanamycin or amikacin. The marker can be used to inactivate or modify specific chloroplast genes, and can be used as a reporter of gene expression. The availability of this marker now makes possible the serial transformation of the chloroplast genome of Chlamydomonas. PMID- 10821175 TI - Protein phosphatase type-1 regulatory subunits Reg1p and Reg2p act as signal transducers in the glucose-induced inactivation of maltose permease in Saccharomyces cerevisiae. AB - The REG1 gene encodes a regulatory subunit of the type-1 protein phosphatase (PP1) G1c7 in Saccharomyces cerevisiae, which directs the catalytic subunit to substrates involved in glucose repression. Loss of REG1 relieves glucose repression of many genes, including the MAL structural genes that encode the maltose fermentation enzymes. In this report, we explore the role of Reglp and its homolog Reg2p in glucose-induced inactivation of maltose permease. Glucose stimulates the proteolysis of maltose permease and very rapid loss of maltose transport activity - more rapid than can be explained by loss of the permease protein alone. In a reg1delta strain we observe a significantly reduced rate of glucose-induced proteolysis of maltose permease, and the rapid loss of maltose transport activity does not occur. Instead, surprisingly, the slow rate of proteolysis of maltose permease is accompanied by an increase in maltose transport activity. Loss of Reg2p modestly reduces the rates of both glucose induced proteolysis of maltose permease and inactivation of maltose transport activity. Overexpression of Reg2p in a reg1delta strain suppresses the effect on maltose permease proteolysis and partially restores the inactivation of maltose transport activity, but does not affect the insensitivity of MAL gene expression to repression by glucose observed in this strain. Thus, protein phosphatase type 1 (Glc7p-Reglp and Glc7p-Reg2p) plays a role in transduction of the glucose signal during glucose-induced proteolysis of maltose permease, but only Glc7p Reglp is involved in glucose-induced inactivation of maltose transport activity and glucose repression of MAL gene expression. Overexpression of REG1 partially restores proteolysis of maltose permease in a grr1delta strain, which lacks glucose signaling, but does not rescue rapid inactivation of maltose transport activity or sensitivity to glucose repression. A model for the role of Reglp and Reg2p in glucose signaling pathways is discussed. We also uncovered a previously unrecognized G2/M delay in the grr1delta but not the reg1delta strains, and this delay is suppressed by REG1 overexpression. The G1/S delay seen in grr1delta mutants is slightly suppressed as well, but REG1 overexpression does not suppress other grr1delta phenotypes such as insensitivity to glucose repression. PMID- 10821176 TI - A carotenogenic gene cluster from Brevibacterium linens with novel lycopene cyclase genes involved in the synthesis of aromatic carotenoids. AB - The carotenogenic (crt) gene cluster from Brevibacterium linens, a member of the commercially important group of coryneform bacteria, was cloned and identified. An expression library of B. linens genes was constructed and a fragment of the crt cluster was obtained by functional complementation of a colourless B. flavum mutant, screening transformed cells for production of a yellow pigment. Subsequent screening of a cosmid library resulted in the cloning of the whole crt cluster from B. linens. All genes necessary for the synthesis of the aromatic carotenoid isorenieratene were identified on the basis of sequence homologies. In addition a novel type of lycopene cyclase was identified by complementation of a lycopene-accumulating B. flavum mutant. Two genes, named crt Yc and crt Yd, which code for polypeptides of 125 and 107 amino acids, respectively, are necessary to convert lycopene to beta-carotene. The amino acid sequences of these polypeptides show no similarity to any of the known lycopene cyclases. This is the first example of a carotenoid biosynthetic conversion in which two different gene products are involved, probably forming a heterodimer. PMID- 10821177 TI - Senescence associated with the over-replication of a mitochondrial retroplasmid in Neurospora crassa. AB - Mitochondrial DNA rearrangements and deletions are a prevailing feature of filamentous fungal cultures that undergo senescence. In Neurospora spp., strains containing the Mauriceville and Varkud mitochondrial retroplasmids routinely senesce at elevated temperatures, a process that is initiated by the integration of variant forms of the plasmids into the mitochondrial genome. Here, we describe a strain that is phenotypically distinguishable from previously characterized senescent strains and show that senescence can occur in the absence of plasmid integration and associated alterations in mitochondrial DNA. The MS4416 strain contains a unique variant of the Mauriceville retroplasmid, and undergoes senescence at highly predictable frequencies at 37 degrees, 25 degrees and 18 degrees C. Decline in vegetative growth rate correlates with increased levels of the variant plasmid and alterations in the synthesis of mitochondrially encoded proteins, suggesting that plasmid over-replication interferes with mitochondrial translation. We also report the isolation of a mutant strain that escapes senescence yet still maintains high levels of the variant plasmid. Its ability to tolerate a growth-suppressive retroplasmid suggests that there are mechanisms in Neurospora which compensate for the deleterious effects that plasmid over replication has on mitochondrial function. PMID- 10821178 TI - P-element transposition in Drosophila melanogaster: influence of size and arrangement in pairs. AB - In Drosophila melanogaster, several factors have been suggested to influence the rates of P-element transposition and excision, including position effects, size and structure of the elements and differences in transposase source. We have investigated the effect of the size of the starting P-element on the rates of excision and transposition. Four transgenes localized at the same insertion site on the X chromosome and which differ by the number of copies of an internal repeated sequence, were studied. Transgenes with sizes ranging from 11 kb to 22 kb excise at similar rates, and size does not correlate with the differences in transposition rate between them. We also studied the behavior of double P elements, located at the same site and arranged in various configurations: nested, contiguous or separated by a few base pairs, in the same or reverse orientation. These double P-elements display different mobilities depending on the arrangement of the two transgenes. Transposition and excision rates were also studied for an insertion bearing four transgenes in very close proximity. Our results suggest that several neighboring elements could excise together. We also propose a new model to explain the formation of all the double P-elements we describe. PMID- 10821179 TI - The isolation and characterization of missense mutants in the general repressor protein Ssn6 of Saccharomyces cerevisiae. AB - Ssn6, a TPR repeat-containing protein, associates with the Tup1 protein to form a general transcriptional repression complex in Saccharomyces cerevisiae. As part of a genetic analysis of this complex, we targeted mutations to the TPR repeat coding region of the SSN6 gene, and applied selection for constitutive expression of the hypoxic gene ANB1. All but one of the resulting mutants failed to express full-length Ssn6 protein, indicating that they harbored deletion, frameshift, or nonsense mutations. The one missense mutation encoded a protein with three amino acid substitutions, and the combination was required for the mutant phenotype. One mutation, a proline substitution for a serine at codon 51, was used in a second round of mutagenesis in which six further multiple-substitution alleles were obtained. These were separated into their component mutations, and again, all but one of the single substitutions displayed the wild-type phenotype. The single and multiple mutants were characterized in terms of their effects on the repression of the glucose-repressible SUC2 gene and the a mating-type gene STE2. The mutant Ssn6 proteins were also tested for their ability to associate with Tup1. The S5 P mutation, despite its lack of a mutant phenotype, had lost the ability to fully associate with Tupl in vitro. In general, those single substitutions that fell within the first two TPR repeats impaired Tup1 associating activity, while the two that fell in TPR repeats five and eight retained this activity. Overexpression of TUP1 partially suppressed the mutant phenotype in only some of the multiple mutants. The results are discussed in terms of the current models of Ssn6 function and the structure of TPR repeats. PMID- 10821180 TI - Characterisation of a cyanide hydratase gene in the phytopathogenic fungus Leptosphaeria maculans. AB - A gene encoding a cyanide hydratase was cloned from an aggressive isolate of Leptosphaeria maculans, the fungus which causes blackleg disease of oilseed Brassica spp. This enzyme catalyses the breakdown of hydrogen cyanide to a less toxic compound, formamide. The predicted amino acid sequence of cyanide hydratase in L. maculans is 77% and 82% identical to cyanide hydratases from two other ascomycetes, Gloeocercospora sorghi and Fusarium lateritium, respectively. The gene is present as a single copy in the L. maculans genome, in both aggressive and non-aggressive isolates, although there is a restriction fragment length polymorphism between these two isolate groups for this gene. The cyanide hydratase promoter contains four putative target sites for GATA transcription factors, proteins that regulate nitrogen metabolism and other processes. Transcription of cyanide hydratase in an aggressive L. maculans isolate is induced strongly by potassium cyanide. Transcription of the gene is detectable in cotyledons of Brassica juncea and B. napus during infection. L. maculans can utilise the reaction product, formamide, as a sole source of nitrogen. PMID- 10821181 TI - Phenotypic and molecular characterization of conjugative antibiotic resistance plasmids isolated from bacterial communities of activated sludge. AB - In order to isolate antibiotic resistance plasmids from bacterial communities found in activated sludge, derivatives of the 3-chlorobenzoate-degrading strain Pseudomonas sp. B13, tagged with the green fluorescent protein as an identification marker, were used as recipients in filter crosses. Transconjugants were selected on agar plates containing 3-chlorobenzoate as the sole carbon source and the antibiotic tetracycline, streptomycin or spectinomycin, and were recovered at frequencies in the range of 10(-5) to 10(-8) per recipient. A total of 12 distinct plasmids, designated pB1-pB12, was identified. Their sizes ranged between 41 to 69 kb and they conferred various patterns of antibiotic resistance on their hosts. Two of the plasmids, pB10 and pB11, also mediated resistance to inorganic mercury. Seven of the 12 plasmids were identified as broad-host-range plasmids, displaying extremely high transfer frequencies in filter crosses, ranging from 10(-1) to 10(-2) per recipient cell. Ten of the 12 plasmids belonged to the IncP incompatibility group, based on replicon typing using IncP group specific PCR primers. DNA sequencing of PCR amplification products further revealed that eight of the 12 plasmids belonged to the IncPbeta subgroup, whereas two plasmids were identified as IncPalpha plasmids. Analysis of the IncP-specific PCR products revealed considerable differences among the IncPbeta plasmids at the DNA sequence level. In order to characterize the gene "load" of the IncP plasmids, restriction fragments were cloned and their DNA sequences established. A remarkable diversity of putative proteins encoded by these fragments was identified. Besides transposases and proteins involved in antibiotic resistance, two putative DNA invertases belonging to the Din family, a methyltransferase of a type I restriction/modification system, a superoxide dismutase, parts of a putative efflux system belonging to the RND family, and proteins of unknown function were identified. PMID- 10821182 TI - Som1, a third component of the yeast mitochondrial inner membrane peptidase complex that contains Imp1 and Imp2. AB - The mitochondrial inner membrane peptidase Imp is required for proteolytic processing of the mitochondrially encoded protein Cox2, the nucleus-encoded Cyt b2, Mcr1, and Cyt c1, and possibly other proteins, during their transport across the mitochondrial membranes. The peptidase contains two catalytic subunits, Imp1 and Imp2. The small protein Soml was previously shown to affect the function of Imp1, but the precise role of Soml remained unknown. Using mutants deleted for IMP1, IMP2 and SOM1, we show here that the Som1 protein is absent in the imp1delta mutant, whereas the level of the Imp1 subunit of the peptidase is only slightly reduced in the soml null mutant. The Soml protein is not essential for proteolytic processing of Cyt b2, while the two other known Imp1 substrates, Cox2 and Mcr1, are not processed in the absence of Som1. Proteolytic processing of Cyt c1 by the Imp2 subunit, and of Ccp by an as yet unidentified peptidase, is not impaired in the som1 deletion mutant. By crosslinking and co-immunoprecipitation assays we demonstrate that the Imp1 and Som1 proteins physically interact. We conclude from our results that stabilisation of Som1 and correct Imp1 function is mediated by a direct interaction between the Imp1 and Som1 proteins, suggesting that Som1 represents a third subunit of the Imp peptidase complex. PMID- 10821183 TI - A nuclear protein that binds specifically to several maize transposons is not essential for Ds1 excision. AB - Specific binding of plant nuclear proteins to GGTAAA-like motifs in the terminal regions of the transposable elements Ac and Mu1 has been detected in several laboratories. However, the role of these proteins in transposition remains unknown. To test the hypothesis that this binding activity is necessary for transposition, we identified and mutagenized all the binding motifs within the Ds1 element. This analysis enabled us to define more precisely the requirements for binding of the host protein. We then tested the ability of the mutated elements to excise from the maize streak virus (MSV) genome. We found that mutated Ds1 elements that do not bind the host proteins, as determined by gel shift competition assay, are still capable of undergoing excision in maize, although for one of the maize lines the rate of excision was reduced. Excision of mutated Ds1 elements generated typical excision footprints. These data indicate that binding of host protein(s) to the GGTAAA-like motifs is not essential for Ds1 excision; however, it may contribute to the efficiency of the process. PMID- 10821184 TI - Genetic analysis of olfC demonstrates a role for the position-specific integrins in the olfactory system of Drosophila melanogaster. AB - Genetic analysis of olfC provides evidence for a role for integrins in the development and/or function of the olfactory system of Drosophila. The olfC gene was identified on the basis of mutations that result in specific defects in behavioural responses to acetate esters, and has been mapped to the cytogenetic interval 7D1;D5-6 on the X chromosome. The myospheroid (mys) gene maps to this region and encodes a beta subunit of integrins. Integrins are alpha(beta) heterodimers which are present on the cell surface and have been implicated in a variety of signalling roles. Mutations in mys fail to complement the olfactory deficits of olfC mutants. These defects can be rescued by misexpression of the mys+ gene under control of a hsp70 promoter. Mutations that affect the alpha subunit of the position-specific integrin PS2 show a dominant interaction with olfC. These results suggest that olfC is allelic to mys and functions together with alphaPS2 integrins in the olfactory pathway in Drosophila. PMID- 10821185 TI - Ambient pH signalling in ascomycetous yeasts involves homologues of the Aspergillus nidulans genes palF and paIH. AB - In Yarrowia lipolytica, the transcription factor Rim101p mediates both pH regulation and control of mating and sporulation. Like its homologues PacC of Aspergillus nidulans and Rim101p of Saccharomyces cerevisiae, Y1Rim101p is activated by proteolytic C-terminal processing, which occurs in response to a signal transduced by a pathway involving several PAL gene products. We report here the cloning and sequencing of two of these genes, PAL2 and PAL3. PAL2 encodes a putative 632-residue protein with six possible transmembrane segments, which differs from the transmembrane proteins Rim9p of S. cerevisiae and Pall of A. nidulans, but is homologous to A. nidulans Pa1H and to the product of the ORF YNL294c, a predicted polypeptide of unknown function in S. cerevisiae. PAL3 encodes an 881-residue polypeptide that is homologous to PalF of A. nidulans and to a newly identified putative polypeptide of S. cerevisiae. Both PAL2 and PAL3 are expressed constitutively, regardless of ambient pH. Mutations in these genes affect growth at alkaline pH and sporulation in both Y. lipolytica and in S. cerevisiae. They affect invasiveness of haploid strains in S. cerevisiae only, and conjugation in Y. lipolytica only. These results highlight the conservation of the Pal pathway initially described in A. nidulans. PMID- 10821186 TI - The mitochondrial DNA of Dictyostelium discoideum: complete sequence, gene content and genome organization. AB - We present an overview of the gene content and organization of the mitochondrial genome of Dictyostelium discoideum. The mitochondria genome consists of 55,564 bp with an A + T content of 72.6%. The identified genes include those for two ribosomal RNAs (rn1 and rns), 18 tRNAs, ten subunits of the NADH dehydrogenase complex (nad1, 2, 3, 4, 4L, 5, 6, 7, 9 and 11), apocytochrome b (cytb), three subunits of the cytochrome oxidase (cox1/2 and 3), four subunits of the ATP synthase complex (atp1, 6, 8 and 9), 15 ribosomal proteins, and five other ORFs, excluding intronic ORFs. Notable features of D. discoideum mtDNA include the following. (1) All genes are encoded on the same strand of the DNA and a universal genetic code is used. (2) The cox1 gene has no termination codon and is fused to the downstream cox2 gene. The 13 genes for ribosomal proteins and four ORF genes form a cluster 15.4 kb long with several gene overlaps. (3) The number of tRNAs encoded in the genome is not sufficient to support the synthesis of mitochondrial protein. (4) In total, five group I introns reside in rnl and cox1/2, and three of those in cox1/2 contain four free-standing ORFs. We compare the genome to other sequenced mitochondrial genomes, particularly that of Acanthamoeba castellanii. PMID- 10821187 TI - The yeast peptidyl proline isomerases FPR3 and FPR4, in high copy numbers, suppress defects resulting from the absence of the E3 ubiquitin ligase TOM1. AB - Tom1p is a 3268-amino acid protein with extensive homology to the hect-domain class of E3 ubiquitin ligases. Disruption of the TOMI gene results in temperature sensitivity for growth. Genes encoding the peptidyl proline isomerases Fpr3p and Fpr4p, when present on multicopy plasmids, will suppress this temperature sensitive growth phenotype. FPR3 can also suppress the mating defect seen in tom1 strains. Suppression is specific for disruption of TOM1, since FPR3 does not restore wild-type growth to strains lacking the E2 ubiquitin-conjugating enzyme Rad6p or the transcriptional regulator Ngglp. Interestingly, the peptidyl proline isomerase domains of Fpr3p and Fpr4p are not required for suppression; rather the essential sequences include about 170 highly conserved residues at the proteins' N-termini. Previously we found that Tomlp plays a role in gene regulation. Since overexpression of FPR4 does not suppress the reduced expression of the ARG1 promoter found in tom1 deletion strains, Tom1p probably has one or more functions beyond its involvement in gene expression. PMID- 10821188 TI - Functional specificity of the mitochondrial DnaJ protein, Mdj1p, in Saccharomyces cerevisiae. AB - Inactivation of the gene for the mitochondrial DnaJ homolog, Mdj1p, in Saccharomyces cerevisiae results in temperature sensitivity and the loss of respiratory activity; the latter phenotype has been attributed to the loss of mitochondrial DNA. To investigate the functional specificity of Mdj1p, non mitochondrial DnaJ proteins were targeted to mitochondria and tested for their ability to substitute for Mdj1p. The tested DnaJ proteins were able to complement the two Mdj1p-linked phenotypes, i.e., respiratory activity and growth at 37 degrees C, to different extents, ranging from full to very poor complementation. All DnaJ homologs ensured faithful propagation of the mitochondrial genome. N terminal fragments of Mdjlp and Escherichia coli DnaJ comprising the well characterized J domain partially substituted for Mdj1p. As the only hitherto known function of the N-terminal fragment is modulation of the substrate binding activity of the cognate Hsp70, we conclude that both Mdj1p-linked phenotypes - maintenance of respiratory activity and the ability to grow at elevated temperature - involve a mitochondrial Hsp70 partner protein. PMID- 10821189 TI - Cysteine is essential for transcriptional regulation of the sulfur assimilation genes in Saccharomyces cerevisiae. AB - Transcription of the genes for sulfur assimilation and methionine biosynthesis in Saccharomyces cerevisiae is regulated by the size of the intracellular pool of an organic sulfur compound. The identity of this compound is not clear, but suggestions include S-adenosylmethionine (SAM) and cysteine. By studying the repression of selected sulfur assimilation (MET) genes, we found that the ability to form cysteine from homocysteine is crucial for methionine-mediated repression to take place. The transcription of MET14 and MET25 could not be repressed by methionine in strains in which either STR4 (which encodes cystathionine beta synthase) or STR1 (cystathionine gamma-lyase) was disrupted, whereas the repression was independent of GSH1 (which encodes the enzyme responsible for the first step in glutathione biosynthesis from cysteine). In contrast, cysteine could repress the MET genes in all of these strains. Two genes that presumably encode cystathionine gamma-synthase and cystathionine beta-lyase were identified by genetic disruption (ORFs YJR130c and YGL184c), yielding yeast strains that cannot convert cysteine into homocysteine. Repression by cysteine was possible in either disruptant, suggesting a role in repression for cysteine alone. While some repression of MET genes could be accomplished by homocysteine in a strain that cannot form SAM from methionine, a low intracellular level of SAM seems to be necessary for full cysteine-mediated repression to take place. PMID- 10821190 TI - Duplicated gene clusters suggest an interplay of glycogen and trehalose metabolism during sequential stages of aerial mycelium development in Streptomyces coelicolor A3(2). AB - DNA sequencing and operon disruption experiments indicate that the genes glgBI and glgBII, which code for the two developmentally specific glycogen branching enzymes of Streptomyces coelicolor A3(2) each form part of larger duplicated operons consisting of at least four genes in the order pep1-treS-pep2-glgB. The sequences of the TreS proteins are 73% identical (93% similar) to that of an enzyme that converts maltose into trehalose in Pinmelobacter, a distantly related actinomycete; and the Pep1 proteins show relatedness to the alpha-amylase superfamily. Disruptions of each operon have spatially specific effects on the nature of glycogen deposits, as assessed by electron microscopy. Upstream of the glgBI operon, and diverging from it, is a gene (glgP) that encodes a protein resembling glycogen phosphorylase from Thermatoga maritima and a homologue in Mycobacterium tuberculosis. These three proteins form a distinctive subgroup compared with glycogen phosphorylases from most other bacteria, which more closely resemble the enzymes from eukaryotes. Diverging from the glgBII operon, and separated from the pep1 gene by two very small ORFs, is a gene (glgX) encoding a probable glycogen debranching enzyme. It is suggested that most of these gene products participate in the developmentally modulated interconversion of immobile, inert glycogen reservoirs, and diffusible forms of carbon, both metabolically active (e.g. glucose-1-phosphate generated by glycogen phosphorylase) and metabolically inert but physiologically significant (trehalose). PMID- 10821191 TI - Rapid isolation of promoter sequences by TAIL-PCR: the 5'-flanking regions of Pal and Pgi genes from yams (Dioscorea). AB - Using a modified TAIL-PCR technique, the 5'-flanking regions of the phenylalanine ammonia lyase (Pal) genes of a yam species, Dioscorea bulbifera, and the phosphoglucose isomerase (Pgi) gene of D. tokoro were successfully isolated. Two novel modifications of the TAIL-PCR procedure introduced here, namely (1) the use of a battery of random 10-mers (RAPD primers) as short arbitrary primers, and (2) the use of a total of five nested, gene-specific primers, allow the rapid isolation of the 5'-flanking region of any gene from organisms with large genomes. Isolated 5'-flanking regions were fused to the gus gene, and tested for transient expression in tobacco BY2 cells. All the isolated 5'-flanking regions were shown to drive reporter gene expression. Three Pal promoters responded to salicylic acid, presumably as a result of the binding of a MYB transcriptional activator to the multiple MREs (Myb Recognition Elements) present in these regions. PMID- 10821192 TI - Focalism: a source of durability bias in affective forecasting. AB - The durability bias, the tendency to overpredict the duration of affective reactions to future events, may be due in part to focalism, whereby people focus too much on the event in question and not enough on the consequences of other future events. If so, asking people to think about other future activities should reduce the durability bias. In Studies 1-3, college football fans were less likely to overpredict how long the outcome of a football game would influence their happiness if they first thought about how much time they would spend on other future activities. Studies 4 and 5 ruled out alternative explanations and found evidence for a distraction interpretation, that people who think about future events moderate their forecasts because they believe that these events will reduce thinking about the focal event. The authors discuss the implications of focalism for other literatures, such as the planning fallacy. PMID- 10821193 TI - Facing faces: studies on the cognitive aspects of physiognomy. AB - Physiognomy, the art of reading personality traits from faces, dates back to ancient Greece, and is still very popular. The present studies examine several aspects and consequences of the process of reading traits from faces. Using faces with neutral expressions, it is demonstrated that personality information conveyed in faces changes the interpretation of verbal information. Moreover, it is shown that physiognomic information has a consistent effect on decisions, and creates overconfidence in judgments. It is argued, however, that the process of "reading from faces" is just one side of the coin, the other side of which is "reading into faces." Consistent with the latter, information about personality changes the perception of facial features and, accordingly, the perceived similarity between faces. The implications of both processes and questions regarding their automaticity are discussed. PMID- 10821194 TI - The psychology of the unthinkable: taboo trade-offs, forbidden base rates, and heretical counterfactuals. AB - Five studies explored cognitive, affective, and behavioral responses to proscribed forms of social cognition. Experiments 1 and 2 revealed that people responded to taboo trade-offs that monetized sacred values with moral outrage and cleansing. Experiments 3 and 4 revealed that racial egalitarians were least likely to use, and angriest at those who did use, race-tainted base rates and that egalitarians who inadvertently used such base rates tried to reaffirm their fair-mindedness. Experiment 5 revealed that Christian fundamentalists were most likely to reject heretical counterfactuals that applied everyday causal schemata to Biblical narratives and to engage in moral cleansing after merely contemplating such possibilities. Although the results fit the sacred-value protection model (SVPM) better than rival formulations, the SVPM must draw on cross-cultural taxonomies of relational schemata to specify normative boundaries on thought. PMID- 10821195 TI - Just say no (to stereotyping): effects of training in the negation of stereotypic associations on stereotype activation. AB - The primary aim of the present research was to examine the effect of training in negating stereotype associations on stereotype activation. Across 3 studies, participants received practice in negating stereotypes related to skinhead and racial categories. The subsequent automatic activation of stereotypes was measured using either a primed Stroop task (Studies I and 2) or a person categorization task (Study 3). The results demonstrate that when receiving no training or training in a nontarget category stereotype, participants exhibited spontaneous stereotype activation. After receiving an extensive amount of training related to a specific category, however, participants demonstrated reduced stereotype activation. The results from the training task provide further evidence for the impact of practice on participants' proficiency in negating stereotypes. PMID- 10821196 TI - Influence of social motives on integrative negotiation: a meta-analytic review and test of two theories. AB - A meta-analysis of 28 studies examined support for the Theory of Cooperation and Competition (M. Deutsch, 1973) and Dual Concern Theory (D. G. Pruitt & J. Z. Rubin, 1986). Effects of social motive (prosocial vs. egoistic) and resistance to yielding (high vs. low vs. unknown) on contenting, problem solving, and joint outcomes were examined. Consistent with Dual Concern Theory, results showed that negotiators were less contentious, engaged in more problem solving, and achieved higher joint outcomes when they had a prosocial rather than egoistic motive, but only when resistance to yielding was high (or unknown) rather than low. The authors also explored the moderating effects of study characteristics and found effects for participation inducement (class exercise, participant pool), for publication status, and for treatment of no-agreement dyads. PMID- 10821197 TI - Pro-norm and anti-norm deviance within and between groups. AB - Participants evaluated other individuals who deviated in either an anti- or pro normative direction relative to normative members. In Study 1, in-group gender normative members were rated more positively than deviant members. The pro-norm deviant was viewed as more attractive than the anti-norm deviant. In Study 2 anti norm in-group deviants were evaluated more negatively than anti-norm out-group deviants even though both held identical attitudes. In both studies, despite objective equivalence, pro-norm deviance was perceived as less "atypical" than anti-norm deviance. Judgments and reactions to deviance depend on group membership and the direction of deviance, not just its magnitude. Evaluations of deviants are also related to perceivers' identification with their own group. These findings are consistent with our model of subjective group dynamics. PMID- 10821198 TI - The moderating effect of trivial triggering provocation on displaced aggression. AB - Two studies examined the interaction between the presence or absence of (a) an initial provocation and (b) a subsequent minor triggering action on the part of the target of displaced aggression. Consistent with the triggering event being seen by participants as indeed trivial when administered by itself without prior provocation, exposure to it literally had no impact on aggression toward its source. When previously provoked, however, this subsequent triggering event strongly increased displaced aggression, causing it to reliably exceed both that displayed when there was no antecedent provocation and that elicited by provocation alone. Mediation analyses showed that for participants who had been provoked, subjective feelings of displeasure concerning the triggering event mediated the effect of the trigger on aggression. PMID- 10821199 TI - The frequency of temporal-self and social comparisons in people's personal appraisals. AB - Although past literature emphasizes the importance of social comparisons, in this study it was predicted that participants would often mention temporal comparisons in their self-descriptions. The first 3 studies revealed that participants report as many or more temporal-past comparisons than social comparisons. It was predicted that people would particularly favor temporal-past comparisons when they are interested in enhancing themselves. Temporal-past comparisons are gratifying, because they tend to indicate improvement over time. Social comparisons may be preferred when people are motivated to evaluate themselves accurately. These predictions were supported when self-evaluation and self enhancement goals were explicitly manipulated (Study 4) or primed (Study 5). PMID- 10821200 TI - Culture, control, and perception of relationships in the environment. AB - East Asian cognition has been held to be relatively holistic; that is, attention is paid to the field as a whole. Western cognition, in contrast, has been held to be object focused and control oriented. In this study East Asians (mostly Chinese) and Americans were compared on detection of covariation and field dependence. The results showed the following: (a) Chinese participants reported stronger association between events, were more responsive to differences in covariation, and were more confident about their covariation judgments; (b) these cultural differences disappeared when participants believed they had some control over the covariation judgment task; (c) American participants made fewer mistakes on the Rod-and-Frame Test, indicating that they were less field dependent; (d) American performance and confidence, but not that of Asians, increased when participants were given manual control of the test. Possible origins of the perceptual differences are discussed. PMID- 10821201 TI - An event-related potential analysis of extraversion and individual differences in cognitive processing speed and response execution. AB - Individual differences in cognitive processing speed and response execution were examined in relation to extraversion. Event-related potentials (ERPs) were recorded concurrently with reaction time and movement time (MT) measures as participants (N = 67) performed simple reaction time and stimulus-response compatibility tasks. Slower processing speed for extraverts, as indicated by longer latency of a late positive ERP wave, P3, was only evident in conditions in which stimulus information was in conflict with response selection demands. As previously reported, the salient effect in all conditions of both tasks was faster MT for extraverts, an effect that is indicative of differences in fundamental motor processes. On the simple reaction time task, amplitudes of the N1 component, an early negative ERP wave, were smaller for extraverts than for introverts in response to auditory tones, an effect that affirms the enhanced sensory reactivity of introverts to punctate physical stimuli. PMID- 10821202 TI - Making judgments about ability: the role of implicit theories of ability in moderating inferences from temporal and social comparison information. AB - Two studies examined the novel proposal that implicit theories of intelligence (C. S. Dweck & E. L. Leggett, 1988) moderate both the effects of performance trends on ability inferences and the perceived diagnosticity of temporal versus normative feedback. Results from 613 adolescents and 42 teachers confirmed that entity theorists perceived initial outcome as more diagnostic and inferred higher ability in another (Study 1) and in the self (Study 2) in a declining outcome condition; incremental theorists perceived last outcome as more diagnostic and inferred higher ability in an ascending condition. Experimental induction of beliefs about ability had similar effects. As predicted, self-appraisal was affected more by temporal feedback among incremental theorists and by normative feedback among entity theorists. Results help resolve prior mixed findings regarding order effects and responses to temporal and normative evaluation. PMID- 10821203 TI - Stress and alcohol use: a daily process examination of the stressor-vulnerability model. AB - The authors used a daily diary methodology to examine over 60 days how the within person associations among event stress, alcohol consumption, and desire to drink varied as a function of gender, positive and negative alcohol-outcome expectancies, and avoidant coping in a sample of 88 regular drinkers. Multilevel regression analyses indicated that men who more strongly anticipated positive outcomes or a sense of carelessness from drinking drank relatively more on stressful days compared with low-stress days. Similar results were found predicting desire to drink. Men who anticipated greater impairment from drinking drank relatively less on stressful days. In general, these effects did not hold for women. Little evidence was found for the predicted effects for avoidant coping style, and some results showed that avoidant coping style buffered the exacerbating effects of careless unconcern expectancies. PMID- 10821204 TI - Differentiating benzodiazepine- and barbiturate-like discriminative stimulus effects of lorazepam, diazepam, pentobarbital, imidazenil and zaleplon in two- versus three-lever procedures. AB - Previous studies found that animals trained to discriminate pentobarbital show a relatively inclusive generalization profile. They generalize to sedative hypnotics and anxiolytics, regardless of differences among such drugs in molecular mechanism of action. In contrast, animals trained to discriminate lorazepam have shown a generalization profile that appears selective for compounds with in-vitro profiles as full agonists at the benzodiazepine modulatory site. The present study investigated whether benzodiazepine receptor ligands, to which pentobarbital-trained rats had generalized under a two-lever procedure, would occasion pentobarbital- or lorazepam-appropriate responding when the rats were retrained to discriminate among pentobarbital, lorazepam and the no drug condition under a three-lever procedure. A second group of rats was trained first to discriminate lorazepam and then retrained under the same three-lever procedure. Under the two-lever procedure, all pentobarbital-trained rats showed dose-dependent generalization to lorazepam, but not all lorazepam-trained rats showed full generalization to pentobarbital. Both groups showed full generalization to diazepam and zaleplon, a novel hypnotic that is selective for alpha, 1-subunit-containing subtypes of the gamma-aminobutyric acid (GABA)A receptor. Pentobarbital-trained rats, but not all lorazepam-trained rats, generalized to imidazenil. Under the three-lever procedure, dose-dependent generalization to lorazepam and pentobarbital was demonstrated on the appropriate levers. Diazepam shared discriminative effects with pentobarbital, zaleplon shared discriminative effects with lorazepam, and imidazenil shared discriminative effects with lorazepam and pentobarbital. These results show that when the opportunity for finer differentiation of discriminative effects of GABAergic drugs is provided, a generalization profile more in line with differential in-vitro profiles can be revealed. PMID- 10821205 TI - The discriminative stimulus effects of morphine in pigeons responding under a progressive ratio schedule of food presentation. AB - The purpose of this study was to determine whether progressive ratio (PR) schedules might provide additional information, as compared with fixed ratio or fixed interval schedules, on pharmacologic features of discriminative stimuli (e.g. stimulus intensity). Five pigeons discriminated between 5.6 mg/kg morphine and saline under an arithmetic PR5 schedule of food presentation. The final ratio before pigeons either stopped responding for 5 min or switched responding from the selected to the non-selected key was designated as the last completed ratio (LCR). Pigeons responded 6.8% on the drug key following saline and 96.4% on the drug key following 5.6 mg/kg morphine. The average LCR value for saline was not significantly different from the average LCR value for morphine. A larger dose of morphine (10.0 mg/kg) increased the LCR value and significantly decreased rates of responding. Smaller doses of morphine (0.32 and 1.0 mg/kg) occasioned primarily saline-appropriate responding and decreased LCR values. Buprenorphine substituted for morphine and significantly increased LCR values, whereas nalbuphine produced only partial (20-80%) morphine-key responding and significantly decreased LCR. Cocaine did not substitute for morphine or modify LCR compared with saline control. Together, these results suggest that the stimulus effects of micro-opioids vary on a dimension (e.g. intensity) that can be quantified using PR schedules. PMID- 10821206 TI - Blockade of the serotonergic system counteracts the dizocilpine-induced changes in dopaminergic function. AB - The administration of dizocilpine, a non-competitive N-methyl-D-aspartate (NMDA) receptor antagonist acting at the associated ion channel, increased the grooming time induced in rats by the D1 dopamine receptor agonist SKF 38393 and the stereotyped behaviour elicited by the D1/D2 dopamine receptor agonist apomorphine, and reduced the locomotor response to the D2 dopamine receptor agonist quinpirole. This supports the view that glutamate deficiency plays an important role in the pathogenesis of schizophrenia by altering the balance between glutamatergic and dopaminergic systems. Blockade of serotonin receptors counteracted the effect of dizocilpine on dopaminergic responses. Both the non selective 5HT1/5HT2 antagonist methysergide, and ketanserin, which more specifically blocks 5HT2 receptors, given at doses inhibiting serotonin-mediated behaviours but which did not affect spontaneous motility and dopaminergic behaviours, hampered the dizocilpine-induced potentiation of responses elicited by the stimulation of D1 or D1/D2 dopamine receptors and counteracted the dizocilpine-induced reduction of hyperactivity observed following quinpirole administration. The results suggest that the functional integrity of the serotonergic system is fundamental for the occurrence of dopaminergic changes resulting from non-competitive NMDA blockade. PMID- 10821207 TI - Effects of N-methyl-D-aspartate receptor antagonists on reinstatement of cocaine seeking behavior by priming injections of cocaine or exposures to cocaine associated cues in rats. AB - The reinstatement of extinguished cocaine self-administration behavior was studied in rats pretreated with N-methyl-D-aspartate receptor antagonists. Rats were trained to self-administer intravenous cocaine (0.32 mg/kg/infusion) during five consecutive daily sessions that were followed by five consecutive daily extinction sessions, during which cocaine was unavailable and cocaine-associated cues (sound and light) were absent. Neither the competitive N-methyl-D-aspartate receptor antagonist D-CPPene (0.3-3 mg/kg) nor the low-affinity N-methyl-D aspartate receptor channel blocker memantine (1-10 mg/kg) reinstated extinguished responding. Priming injections of intravenous cocaine (Experiment 1), and exposures to cocaine-associated stimuli (buzzer and light; Experiment 2) engendered responding on the reinforced lever in excess of that on the non reinforced lever. In Experiment 1, administration of D-CPPene or memantine prior to the priming injection of cocaine eliminated the difference between reinforced lever and non-reinforced-lever response rates. For both D-CPPene and memantine, however, this effect was largely due to increased responding upon the non reinforced lever rather than to decreased reinforced-lever responding. In Experiment 2, D-CPPene, but not memantine, abolished in a dose-dependent manner the selective increase in reinforced-lever over non-reinforced-lever responding that was induced by exposures to cocaine-related stimuli. This effect of D-CPPene was not due to increased non-reinforced-lever responding. These data help define the boundaries within which N-methyl-D-aspartate receptor antagonists can prevent reinstatement of cocaine-seeking behavior (e.g. type of antagonist used and reinstatement procedure). PMID- 10821208 TI - Specificity of effects of chronically administered diazepam on the responding of rats under two different spaced-responding schedules. AB - Lever pressing of rats was maintained in different chambers during two different sessions each day. At 0900 h, responding was maintained under a two-component multiple schedule in which responses initiated an interval that had to elapse before delivery of food (time delay of 20 s and 40 s). In this schedule, a 'response-pause' sequence preceded reinforcers, and acutely administered diazepam only decreased responding. At 1400 h, responding by the same subjects was maintained under a different two-component multiple schedule, in which individual responses initiated an interval that had to be terminated by another response before delivery of food (DRL 20 s and 40 s). In this second schedule, a 'response pause-response' sequence preceded reinforcers, and acutely administered diazepam increased responding. After studying the acute behavioral effects of diazepam during each separate 'timing' schedule, animals systematically received 1.7 mg/kg per day diazepam 2-5 min prior to their different schedule components, in order to study the influence of reinforcement contingency on the chronic effects of this drug. Diminution of the initial effects of diazepam during daily drug administration prior to DRL 20 s responding did not extend to DRL 40 s responding or to time-delay responding, and tolerance did not develop at all for time-delay responding. When diazepam was again administered after all the daily schedules for approximately 1 month, and then given before the individual DRL schedules, DRL responding was increased again as it had been prior to chronic drug administration. These results suggest that the behavioral effects of acutely administered diazepam are influenced by different 'timing' requirements, and that the behavioral effects of chronically administered diazepam are influenced by 'timing' requirements and by drug- and chamber-related stimuli. PMID- 10821209 TI - Intrahippocampal spermidine administration improves inhibitory avoidance performance in rats. AB - Polyamines are polycations present at high concentrations in the mammalian brain. We investigated the effect of an intrahippocampal infusion of spermidine, a polyamine agonist, immediately post-training on the inhibitory avoidance learning paradigm in rats. Bilateral intrahippocampal microinjection of spermidine (0.02 20 nmol) caused an increase in test step-down latencies at low concentrations. Administration of arcaine (0.002-0.2 nmol), an antagonist of the N-methyl-D aspartate (NMDA) receptor polyamine binding site, did not modify the test step down latencies. In contrast, co-administration of arcaine and spermidine completely reversed the facilitatory effect of spermidine on the test step-down latencies. These results provide evidence that polyamines may be involved in learning and memory modulation. PMID- 10821210 TI - Sex differences in the acute effects of cigarette smoking on the reinforcing value of alcohol. AB - Alcohol consumption acutely increases smoking behavior, but the reverse relationship, the acute effects of smoking on alcohol intake, largely has been ignored. We examined whether smoking acutely increases the reinforcing value of alcohol, first in the absence of recent alcohol intake and then following an alcohol pre-load. Healthy, social-drinking smokers (n = 11 men, 14 women) engaged in a computerized task involving concurrent schedules of reinforcement for beer (FR10, 3 oz (90 ml) per reinforcement) or money (FR5 to FR30, $0.20 per reinforcement) during two sessions, one following day-long ad lib smoking and the other following overnight smoking abstinence. During each session, subjects performed the task in two sets of trials, one before and one after consumption of an alcohol pre-load, with 60 min between sets. To standardize the alcohol pre load, all subjects were led to believe that they had earned 9 oz (270 ml) of beer after the first trial set, which they then consumed before the second set of trials. Compared to responding during the abstinent session, responding for alcohol during the smoking session was no different before the alcohol pre-load (trial set one) but was significantly greater following the alcohol pre-load (trial set two), although only in men and not women. Subjective sedation after the alcohol pre-load was attenuated during the smoking session in both men and women, but attenuated sedation due to smoking was related to subsequent alcohol reinforced responding only in men. Additional research is needed to determine the extent to which these effects in men are pharmacological in nature or are conditioned responses to smoking or to consuming a preferred alcoholic beverage. PMID- 10821211 TI - Nicotine gum as a substitute for cigarettes: a behavioral economic analysis. AB - The present experiment attempted to identify a substitute for cigarette smoking in a laboratory analog of the behavioral economics of drug dependence. The interaction between cigarette consumption and nicotine gum consumption was examined with eight human smokers by increasing the price of cigarettes (i.e. the number of responses required to obtain puffs) across sessions, while the price of concurrently available nicotine gum remained constant. In another phase, the price of nicotine gum was increased while the price of concurrently available cigarettes remained constant. To determine whether the presence of concurrently available nicotine gum influenced cigarette consumption, we also examined the effect of increasing the price of cigarettes on cigarette consumption without available nicotine gum. When cigarettes and nicotine gum were concurrently available, increases in the price of cigarettes increased consumption of nicotine gum. When the price of nicotine gum increased while the price of cigarettes remained constant, smokers nearly exclusively consumed cigarettes. The presence or absence of nicotine gum did not affect the relation between cigarette consumption and cigarette price. The results suggest that nicotine gum can maintain operant behavior of smokers in the laboratory and can function, in a behavioral economic sense, as a weak substitute for cigarette puffs. As a result, nicotine gum may be useful in human laboratory studies of the behavioral economics of reinforcer interactions and their role in drug dependence. PMID- 10821212 TI - Delta9-THC training dose as a determinant for (R)-methanandamide generalization in rats: a systematic replication. AB - Jarbe et al. (1998a) trained rats to discriminate between (-)-delta9 tetrahydrocannabinol (delta9-THC) and vehicle, using different training doses in order to create assays with different efficacy demands, to examine whether (R) methanandamide, an analog of the endogenous ligand anandamide, had lower efficacy than delta9-THC. Rats were initially trained with 3 mg/kg delta9-THC, then tested with (R)-methanandamide and delta9-THC. Thereafter, the rats were split into two groups and retrained with either 1.8 or 5.6 mg/kg delta9-THC, followed by additional tests with the two agonists. The current study systematically replicated this study in two groups of rats, trained from the outset to discriminate between vehicle and either 1.8 or 5.6 mg/kg delta9-THC, respectively. Two-lever operant drug discrimination procedures were used. The outcomes in the two studies were similar. In tests with (R)-methanandamide, full substitution occurred in the low-dose delta9-THC training group, whereas substitution was partial in the high-dose delta9-THC training group. (R) Methanandamide in higher doses exerted marked suppression of lever pressing. In tests with delta9-THC, full substitution occurred in both delta9-THC-trained groups, and rates of responding were comparable to those observed during regular drug training sessions. In conclusion, both sets of data indicate that cannabinoid agonists either can have varying degrees of efficacy at a receptor site, or may produce their behavioral actions through multiple mechanisms, or both. Prevailing training-dose condition rather than prior training-dose history is the major determinant for the substitution pattern. PMID- 10821213 TI - Alternative reinforcers differentially modify cocaine self-administration by humans. AB - Six experienced cocaine smokers (two men, four women) participated in an inpatient study to compare self-administration of smoked cocaine when either a $5 money or merchandise voucher was available as an alternative reinforcer. A six trial choice procedure was used, with sessions consisting of (1) one sample trial, where participants received the cocaine dose and the alternative reinforcer available that day, and (2) five choice trials, where participants chose between the available cocaine dose and the alternative reinforcer. There were eight sessions: in separate sessions, each dose of cocaine (0, 12, 25, 50 mg) was paired with a money voucher and with a merchandise voucher. The choice to self-administer cocaine significantly increased with escalating cocaine doses, and significantly less cocaine was self-administered when money vouchers were available as compared to merchandise vouchers. These data demonstrate that money vouchers are a more effective alternative reinforcer than merchandise vouchers in cocaine abusers. PMID- 10821214 TI - Acquisition deficit and time-dependent retrograde amnesia for contextual fear conditioning in agmatine-treated rats. AB - Cumulative evidence indicates that the hippocampus plays a time-limited role in contextual learning paradigms. Pharmacological studies have indicated that acquisition of background contextual cues during Pavlovian fear conditioning is dependent upon hippocampal function, whereas early inactivation of the hippocampus after training produces retrograde amnesia. When administered prior to contextual fear conditioning, agmatine (5 and 10 mg/kg, i.p.), an endogenous polyamine and N-methyl-D-aspartate (NMDA) receptor ligand found at excitatory synapses in the hippocampus, impaired the acquisition of contextual fear (measured as defensive freezing 26 hours later) without a reduction in baseline motor activity during training. Furthermore, ascending doses of agmatine were found not to exert analgesic effects on response thresholds to peripheral shock. This negated the possibility that the observed learning deficit resulted from a difference in perceived shock intensity. Post-training agmatine treatment produced a time-dependent impairment of consolidation, with subjects approaching a level of fear equivalent to that of a reference group as the delay of treatment increased (up to 6 hours). Since physiologically high levels of agmatine are able to inhibit NMDA receptor activity, these results suggest that polyamine modulation of NMDA receptors, most likely within the hippocampus, is required for the acquisition and consolidation of contextual fear stimuli. PMID- 10821215 TI - Melatonin and sex hormone interrelationships--a review. AB - Melatonin, the main hormone secreted by the pineal gland at night, plays a major role in regulating reproductive physiology in seasonal breeders and influences the age of sexual maturation in laboratory rodents. In humans these relationships are less clear. Evidence supporting a melatonin-reproductive hormone relationship relies on findings of abnormal melatonin secretion in disorders of the reproductive system and on pathologies of the pineal gland which are associated with clinical abnormalities of the reproductive hormones. Normal melatonin rhythms are closely related to those of the reproductive hormones during infancy and reciprocally correlated during puberty. The demonstration of melatonin receptors in the brain and in reproductive organs, together with the localization of sex hormone receptors in the pineal gland, further strengthen these relationships. However, it is not yet clear that these correlations are functionally related, as data on the antigonadal effects of exogenous melatonin on the reproductive hormones are not conclusively established. PMID- 10821216 TI - The molecular background of glycogen metabolism disorders. AB - The molecular pathology of classical glycogen storage disorders, glycogen synthase deficiency and Fanconi-Bickel syndrome is reviewed. The isolation of the respective cDNAs, the chromosomal localization of the genes and the elucidation of the genomic organization enabled mutation analysis in most disorders. The findings have shed light on the multi-protein structure of the glucose-6 phosphatase system, the phosphorylase kinase enzymatic complex and the molecular background of the differential tissue expression in debranching enzyme deficiency. The immediate practical benefit of these studies is our extending ability to predict the outcome of clinical variants and to offer genetic counseling to most families. The elucidation of the tertiary structure of these proteins and their structure-function relationship poses major challenges for the future. PMID- 10821217 TI - Gps mutations in Chilean patients harboring growth hormone-secreting pituitary tumors. AB - Hypersecretion of GH is usually caused by a pituitary adenoma and about 40% of these tumors exhibit missense gsp mutations in Arg201 or Gln227 of the Gs, gene. We studied 20 pituitary tumors obtained from patients with GH hypersecretion. One tumor was resected from an 11 year-old boy with a 3 year history of accelerated growth, associated with increased concentrations of serum GH and IGF-I, which were not suppressed by glucose administration. The remaining 19 tumors were obtained from adult acromegalic patients, who had elevated baseline serum GH levels that did not show evidence of suppression after administration of glucose. The gsp mutations were studied by enzymatic digestion of the amplified PCR fragment of exon 8 (Arg201) and exon 9 (Gln227) with the enzymes NlaIII and NgoAIV, respectively. The tumors obtained from the boy and from nine of the 19 patients with acromegaly exhibited the gsp mutation R201H. None of the tumors had the Gln227 mutation. The gsp positive patients tended to be older, had smaller tumors, and had preoperative basal serum GH levels which were significantly lower (21 +/- 6 vs 56 +/- 16 microg/l, p<0.05) than the gsp negative patients. In this study, we documented the presence of a gsp mutation in Arg201 in a boy with gigantism and in approximately half of 19 Chilean adult patients with acromegaly, similar to other populations. PMID- 10821218 TI - Clinical, cytogenetic and molecular analysis of three 46,XX males. AB - Cytogenetic analysis, fluorescent in situ hybridization (FISH) and polymerase chain reaction (PCR) were applied to characterize the Y-chromosomal breakpoints of three XX male patients. Two of these patients show a breakpoint within a protein kinase gene, PRKY, previously described as a hotspot of ectopic recombination between homologous regions on X and Y chromosomes during male meiosis. The slightly different clinical phenotypes of the three patients cannot be correlated with the localization of the breakpoints. PMID- 10821219 TI - Month of birth and subsequent development of type I diabetes (IDDM). AB - The aim of this study was to find out whether there is a seasonal pattern in the month of birth of children with IDDM in Israel and whether this pattern, if present, differs from that of total live births. One thousand and ninety-five out of 1,188 children and adolescents (0-17 years) who developed IDDM in Israel between 1980-1993 and whose month of birth was known were included in the study. Separate analysis was made for Jews (n = 987) who have a high incidence (10 18/105) and Arabs (n = 108) with a low incidence (2.9/105) of IDDM. The pattern of total live birth distribution in Israel over a 20-year period served as control. A significantly different seasonal pattern was found in the two IDDM populations. Whereas the Jews had the lowest number of births in winter (January March) and the highest in spring (April-June), the Arabs presented an inverse pattern in the first months of the year. The distribution of births of the children who developed IDDM was different from that observed in total live births in the Jewish population. The findings in the Jewish population in Israel (a high incidence group) support the hypothesis that IDDM is triggered in some children by viral infections transmitted by the mother during pregnancy or in the early postnatal period. In the Arab population (a low incidence group) a protective (immune and/or genetic) mechanism may exist. PMID- 10821220 TI - Metabolic control in children and adolescents with diabetes: experience of two Italian Regional Centers. AB - A survey on glycemic control in 201 diabetic children and adolescents, aged 0-18 years, was performed in two Italian Centers for Childhood Diabetes (Chieti and Parma). Mean HbA1c was 7.8+/-1.4 (range 4.8-13.3%; median 7.6%). With progression of diabetes duration, deterioration of glycemic control was observed (r=0.20; p< 0.002), more evident in girls than in boys, with peaks at 14 (8.9+/-2.0 vs 6.9+/ 1.7%; p<0.05) and 16 years (9.5+/-1.4% vs 8.1+/-1.1; p<0.02). No differences were found in BMI values in boys or girls, or for insulin doses which were increased significantly in both sexes according to age (r= 0.33, p<0.04). The number of insulin injections did not influence glycemic control. Only one severe hypoglycemic episode was reported during the period of observation. This study demonstrates that modern management, continuous education and patient and family empowerment are effective in attaining excellent glycemic control without increasing the risk of hypoglycemia. PMID- 10821221 TI - Prevalence of microvascular and neurologic abnormalities in a population of diabetic children. AB - One hundred and twenty-nine (87%) of a total county population of 150 eligible diabetic children together with 144 age- and sex-matched control children participated in a longitudinal, epidemiological study of the evolution of diabetic microvascular disease. At enrollment the median (range) age of the diabetic children was 12.5 (3.7-16.8) years with a median diabetes duration of 2.9 (0.1-13.4) years and a median HbAl of 11.1 (6.8-17.9)%. Two sets of measurements were made over a period of 18 months for all indices of microvascular disease, while autonomic function was studied on one occasion. Urinary albumin excretion in diabetic children was assessed from all voidings during two timed 48-h urine collections and was expressed as urinary albumin/creatinine ratios (ACR). Blood pressure (BP) was measured using a random zero sphygmomanometer. Autonomic function was assessed by pupillary adaptation in darkness, using a portable Polaroid pupillometer, and by heart rate (HR) variation recorded by dedicated computer. Vibration sensation thresholds (VST) (as indices of peripheral neuropathy) were recorded using a Biothesiometer. Limited joint mobility (LJM) was assessed by the "prayer sign". Five (3.9%) diabetic children presented raised mean ACR in more than two of four 24-h urine collections. Fourteen (10.8%) diabetic children were identified as having persistently raised BP during both study periods. Impaired HR response in one HR test was observed in 20 (15.5%) diabetic children, while ten (7.7%) diabetic children demonstrated abnormalities in two or more HR tests. Reduced pupillary adaptation in darkness was found in eight (7.9%) diabetic children. Persistent vibration sensation impairment (VST) in lower limbs was detected in eight (6.2%) diabetic children, while LJM was present in 12 (9.3%) diabetic children. Eight of the 129 diabetic children (6.2%) were found to have abnormality in two and one in three indices of microvascular and autonomic function. Six of nine children had coexistence of impaired autonomic neuropathy and nephropathy. These nine children were diagnosed at a younger age than the rest of the diabetic population (5.1 vs 8.0 yr, p=0.002). Four of nine were aged >11 yr and five of nine had had diabetes for >5 yr. Thus, a constellation of microvascular and neurological abnormalities were demonstrable in a small proportion of diabetic children, who were younger than the rest of the population at the time of onset of their disease. Longitudinal study of this population will demonstrate the clinical significance of these findings. PMID- 10821222 TI - Heart rate variability and cardiovascular tests in young patients with diabetes mellitus type 1. AB - The aim of this study was to obtain information about parameters of heart rate variability (HRV) in three frequency bands (high frequency - HF, low frequency - LF and very low frequency - VLF), the sensitivity of cardiovascular tests, and subjective feelings depending on autonomic nervous system balance in a group of young patients with diabetes mellitus type 1 (IDDM). Sixty-four subjects were examined: 32 patients with diabetes with a mean age of 16.1 +/- 0.7 years and a mean duration of IDDM of 6.3 +/- 0.8 years, and 32 healthy controls matched for age, sex and BMI. Shorter R-R intervals and abnormal values reflecting HRV were found in the diabetic group. In particular, parameters of total power, and HF and LF bands were reduced. The ratio VLF/HF power revealed predominance of sympathetic tone in the diabetic subjects. Although relative power VLF was increased in the supine position, the reactive rise of the VLF band activity in orthostasis was lower in the IDDM group. Using cardiovascular tests (deep breathing, Valsalva, orthostasis), significant differences in reactions were found only in the deep breathing test. Evaluation of sympathetic:parasympathetic:indifferent subjective feelings by questionnaire did not reveal any differences between the diabetic and healthy groups. PMID- 10821223 TI - Children with coeliac disease and insulin dependent diabetes mellitus--growth, diabetes control and dietary intake. AB - We aimed to assess the growth, diabetes control, dietary intake and compliance with a gluten-free diet in children with insulin dependent diabetes mellitus (IDDM) and coeliac disease in a major paediatric and adolescent diabetes clinic. Children with IDDM and biopsy-proven coeliac disease aged <18 years were included and compared with IDDM controls matched for age, sex and duration of diabetes. Twenty patients with coeliac disease and IDDM participated (15 female, age 7.4 17.3 yr), with two matched IDDM controls for each (age 6.9-17.4 yr). The prevalence of coeliac disease in this diabetes clinic population was 2.6%. All patients completed a 3 day food record (3DFR) and a 7 day food frequency questionnaire (FFQ) to assess dietary intake and gluten-free compliance. Diabetes control measured by HbA1c was not different between groups or compared to the overall clinic population (8.48 +/- 0.98% for coeliac patients vs 8.87 +/- 1.46 for IDDM controls vs 8.60 +/- 1.30 for overall clinic population aged 5.0-17.9 yr). Height, weight and BMI standard deviation scores were not different between coeliac patients and IDDM controls. No clinically significant differences were found in intake of energy, macronutrients or micronutrients. The proportion of energy intake from carbohydrate, protein and fat was within recommended ranges, except for a higher saturated fat intake. Only 30% of coeliac patients complied with a strict gluten-free diet, but growth parameters were unaffected by dietary compliance. Thus, we found that children and adolescents with coexisting IDDM and coeliac disease have normal growth, equivalent diabetes control and no differences in energy or nutrient intake compared to matched IDDM controls in our clinic population. PMID- 10821224 TI - Determination of body composition in African-American children: validation of bioelectrical impedence with dual energy X-ray absorptiometry. AB - Body compositional differences between Black and White adults are well-known. It has become increasingly apparent that these racial variations may begin in childhood. Previously, our group validated tetrapolar bioelectrical impedance (BIA) measurements against H2(18O) dilution method to develop prediction formulas of fat free mass (FFM) in healthy White-American children: FFM = 0.524 Ht2/R + 0.415 Wt 0.32. In the present study we used BIA to establish a FFM prediction equation for forty African-American children (19 males and 21 females). Of the females, six were diagnosed with polycystic ovary syndrome (PCOS) and were obese. FFM was determined by dual energy X-ray absorptiometry (DEXA). Impedence measurements by BIA showed a strong correlation with FFM determined by DEXA. In healthy Black children, FFM = 0.84 Ht2/R + 1.10 with a standard error of estimate (SEE) of 1.47 kg (R2 = 0.97). In Black females with PCOS, FFM = 0.62 Ht2/R + 0.21 Wt - 1.94 with a SEE of 1.43 kg (R2 = 0.99). The observed differences in the prediction equations of FFM between White-American and African-American children underline the importance of using race-specific formulas in evaluating body composition. With the overall increase in rates of childhood obesity and more so in the Black race, BIA is an easy and useful tool for the assessment and follow up of body compositional changes with lifestyle interventions. PMID- 10821225 TI - Hypothalamic-pituitary dysfunction associated with Moyamoya disease in children. AB - Moyamoya disease is a rare cerebral vascular disease that results in narrowing of the vessels of the circle of Willis and the formation of a network of collateral vessels at the base of the brain for compensatory perfusion. Moyamoya disease has the highest incidence during the first decade of life, and children present most frequently with transient ischemic attacks. We present two cases of Moyamoya disease in children with associated hypothalamic-pituitary dysfunction. Both children presented to the endocrinologist for decreased growth velocity. One child had hypothyroidism and both had growth hormone deficiency. A review of the literature reveals a few isolated case reports of hypothalamic-pituitary dysfunction occurring with Moyamoya disease and with other states of cerebral vascular insufficiency. We suggest that children with compromise of cerebral vascular perfusion be monitored closely for clinical signs and symptoms of hypothalamic-pituitary dysfunction. PMID- 10821226 TI - Recurrence of a nonsense mutation in the conserved domain of SRY in a Brazilian patient with 46,XY gonadal dysgenesis. AB - We tested a female patient with 46,XY karyotype and pure gonadal dysgenesis for the presence of the SRY gene and for mutations within the SRY conserved domain. A point mutation was identified at nucleotide position 209 with respect to the first ATG. The base substitution is a G-->A transition in the first nucleotide of codon 70 which changes a tryptophan (TGG) to a stop codon (TAG). Even though the father was not available for investigation we assumed that it is a de novo mutation, since it probably generates a nonfunctional truncated protein. PMID- 10821227 TI - Central precocious puberty in 48,XXYY Klinefelter syndrome variant. AB - We report the first case of central precocious puberty in a patient with 48,XXYY Klinefelter syndrome variant. We also report clinical characteristics, growth pattern, endocrine data and pathological testicular findings. The patient did not receive medical care for his precocious pubertal development, because of adequate height prognosis, and reached normal height for both his target height and Klinefelter patients. Since precocious puberty seems to occur in Klinefelter syndrome and its variants, we advise karyotype analysis in boys with mental retardation, gynecomastia, small testes and precocious onset of puberty. PMID- 10821228 TI - Growth failure associated with the use of high dose prozac (fluoxetine hydrochloride) in a patient with type 1 diabetes mellitus. PMID- 10821229 TI - Temporal bone studies of the human peripheral vestibular system. Normative vestibular hair cell data. AB - Quantitative studies of the vestibular system with serially sectioned human temporal bones have been limited because of difficulty in distinguishing hair cells from supporting cells and type I from type II hair cells. In addition, there is only a limited amount of normative data available regarding vestibular hair cell counts in humans. In this study, archival temporal bone sections were examined by Nomarski (differential interference contrast) microscopy, which permitted visualization of the cuticular plate and stereociliary bundle so as to allow unambiguous identification of hair cells. The density of type I, type II, and total numbers of vestibular hair cells in each of the 5 sense organs was determined in a set of 67 normal temporal bones that ranged from birth through 100 years of age. The mean total densities at birth were 76 to 79 cells per 0.01 mm2 in the cristae, 68 cells per 0.01 mm2 in the utricle, and 61 cells per 0.01 mm2 in the saccule. The ratio of type I to type II hair cells at birth was 2.4:1 in the cristae and 1.3:1 in the maculae. There was a highly significant age related decline in all sense organs for total, type I, and type II hair cell densities that was best fit by a linear regression model. The cristae lost type I cells with advancing age at a significantly greater rate than the maculae, whereas age-related losses for type II cells occurred at the same rate for all 5 sense organs. Hair cell densities in the cristae were significantly higher at the periphery than at the center. There were no significant sex or interaural differences for any of the counts. Mathematical models were developed to calculate the mean and 95% prediction intervals for the total, type I, and type II hair cell densities in each sense organ on the basis of age. There was overall good agreement between the hair cell densities determined in this study and those reported by others using surface preparation techniques. Our data and related models will serve as a normative database that will be useful for comparison to counts made from subjects with known vestibular disorders. PMID- 10821230 TI - Temporal bone studies of the human peripheral vestibular system. Normative Scarpa's ganglion cell data. AB - Scarpa's ganglion cell counts were performed in 106 serially sectioned, normal human temporal bones from 75 individuals. Of these 106 bones, 15 were from neonates less than 30 days old, 14 were from infants between the ages of 1 and 12 months, and the remainder were distributed throughout each decade of life, with sample sizes ranging from 4 to 10 per decade. All temporal bones had to meet 2 criteria: no symptoms or signs of inner ear disease except for presbycusis in the medical case history and no abnormality in the inner ear on light microscopy. The total ganglion cell counts declined significantly with age at an average rate of 57 cells per year. The age-related decline was significantly greater in the superior division than in the inferior division. There was also a significant sex effect, independent of age: for any age, the count in men averaged 1,526 cells higher than in women. There was no significant interaural difference. Mathematical models were developed to compute the mean and 95% prediction intervals for Scarpa's ganglion cell counts in terms of age and sex parameters. The counts and models will serve as a normative database against which to compare counts made in temporal bones from subjects with known vestibular disorders. PMID- 10821231 TI - Temporal bone studies of the human peripheral vestibular system. Aminoglycoside ototoxicity. AB - Quantitative assessments of vestibular hair cells and Scarpa's ganglion cells were performed on 17 temporal bones from 10 individuals who had well-documented clinical evidence of aminoglycoside ototoxicity (streptomycin, kanamycin, and neomycin). Assessment of vestibular hair cells was performed by Nomarski (differential interference contrast) microscopy. Hair cell counts were expressed as densities (number of cells per 0.01 mm2 surface area of the sensory epithelium). The results were compared with age-matched normal data. Streptomycin caused a significant loss of both type I and type II hair cells in all 5 vestibular sense organs. In comparing the ototoxic effect on type I versus type II hair cells, there was greater type I hair cell loss for all 3 cristae, but not for the maculae. The vestibular ototoxic effects of kanamycin appeared to be similar to those of streptomycin, but the small sample size precluded definitive conclusions from being made. Neomycin did not cause loss of vestibular hair cells. Within the limits of this study (maximum postototoxicity survival time of 12 months), there was no significant loss of Scarpa's ganglion cells for any of the 3 drugs. The findings have implications in several clinical areas, including the correlation of vestibular test results to pathological findings, the rehabilitation of patients with vestibular ototoxicity, the use of aminoglycosides to treat Meniere's disease, and the development of a vestibular prosthesis. PMID- 10821232 TI - Temporal bone studies of the human peripheral vestibular system. Meniere's disease. AB - Quantitative assessments of vestibular hair cells and Scarpa's ganglion cells were performed on temporal bones from 24 patients with well-documented Meniere's disease. Of these, 18 had unilateral disease and 6 had bilateral disease. Vestibular hair cell counts were made in each of the 5 sense organs by Nomarski (differential interference contrast) microscopy. Hair cell counts were expressed as densities: number of cells per 0.01 mm2 surface area of the sensory epithelium. The results were compared with age- and sex-matched normal data. The type I hair cell densities for all vestibular sense organs were within the range for normative data. On the other hand, there was a significant loss (p < .01) of type II hair cells for all 3 cristae and both maculae. There was also a significant loss of Scarpa's ganglion cells (p < .001) when compared with normative data. The findings indicate a selective loss of type II hair cells and Scarpa's ganglion cells in Meniere's disease. These new observations have implications regarding the pathophysiological mechanism and clinical manifestations of Meniere's disease. PMID- 10821233 TI - Avoid wacky (genome) races. PMID- 10821234 TI - A global solution to global problems? PMID- 10821235 TI - South Africa turns to research in the hope of settling AIDS policy. PMID- 10821236 TI - Editor defends publishing key AZT paper. PMID- 10821237 TI - French research minister targets IT and biotech. PMID- 10821238 TI - German Greens go cold on nuclear fusion PMID- 10821239 TI - Canada's plans for neutrons stall. PMID- 10821240 TI - US reforms rules for telling public about GM food. PMID- 10821242 TI - Israel debates raising commitment to CERN. European Laboratory for Particle Physics PMID- 10821241 TI - Italian genomics boost retained. PMID- 10821243 TI - Troubled US fusion project receives cash injection PMID- 10821244 TI - Medical tests cost Lawrence Berkeley $2.2 million. PMID- 10821245 TI - Australia improves its AIMS. Australian Institute of Marine Science. PMID- 10821246 TI - Souped-up search engines. PMID- 10821247 TI - We urgently need more data to improve the lives of laboratory animals. PMID- 10821248 TI - Planck family paid a high price for opposing Hitler. PMID- 10821249 TI - How to make diplomats scientifically literate PMID- 10821250 TI - Science's neglected legacy. PMID- 10821251 TI - Horse power. PMID- 10821252 TI - Disextinction, Inc PMID- 10821253 TI - Designed to dissolve PMID- 10821255 TI - Cosmology. The dark side of distortion PMID- 10821254 TI - Hearing. Tuning in with motor proteins. PMID- 10821256 TI - The rise of the hybrid fungi. PMID- 10821257 TI - Statistical mechanics. Exploring phase space PMID- 10821259 TI - Lie detection and language comprehension. PMID- 10821258 TI - Gene expression. Mutant weed breaks silence. PMID- 10821260 TI - Formation of endothelial cell networks. PMID- 10821261 TI - Chernobyl's legacy in food and water. PMID- 10821262 TI - Detection of weak gravitational lensing distortions of distant galaxies by cosmic dark matter at large scales AB - Most of the matter in the Universe is not luminous, and can be observed only through its gravitational influence on the appearance of luminous matter. Weak gravitational lensing is a technique that uses the distortions of the images of distant galaxies as a tracer of dark matter: such distortions are induced as the light passes through large-scale distributions of dark matter in the foreground. The patterns of the induced distortions reflect the density of mass along the line of sight and its distribution, and the resulting 'cosmic shear' can be used to distinguish between alternative cosmologies. But previous attempts to measure this effect have been inconclusive. Here we report the detection of cosmic shear on angular scales of up to half a degree using 145,000 galaxies and along three separate lines of sight. We find that the dark matter is distributed in a manner consistent with either an open universe, or a flat universe that is dominated by a cosmological constant. Our results are inconsistent with the standard cold-dark matter model. PMID- 10821264 TI - Cosmic gamma-ray background from structure formation in the intergalactic medium AB - The Universe is filled with a diffuse background of gamma-ray radiation, the origin of which remains one of the unsolved puzzles of cosmology. Less than one quarter of the gamma-ray flux can be attributed to unresolved discrete sources, such as active galactic nuclei; the remainder appears to constitute a truly diffuse background. Here we show that the shock waves induced by gravity in the gas of the intergalactic medium, during the formation of large-scale structures like filaments and sheets of galaxies, produce a population of highly relativistic electrons. These electrons scatter a small fraction of the cosmic microwave background photons in the local Universe up to gamma-ray energies, thereby providing the gamma-ray background. The predicted diffuse flux agrees with the observed background across more than four orders of magnitude in photon energy, and the model predicts that the gamma-ray background, though generated locally, is isotropic to better than five per cent on angular scales larger than a degree. Moreover, the agreement between the predicted and observed background fluxes implies a mean cosmological density of baryons that is consistent with Big Bang nucleosynthesis. PMID- 10821263 TI - Prestin is the motor protein of cochlear outer hair cells. AB - The outer and inner hair cells of the mammalian cochlea perform different functions. In response to changes in membrane potential, the cylindrical outer hair cell rapidly alters its length and stiffness. These mechanical changes, driven by putative molecular motors, are assumed to produce amplification of vibrations in the cochlea that are transduced by inner hair cells. Here we have identified an abundant complementary DNA from a gene, designated Prestin, which is specifically expressed in outer hair cells. Regions of the encoded protein show moderate sequence similarity to pendrin and related sulphate/anion transport proteins. Voltage-induced shape changes can be elicited in cultured human kidney cells that express prestin. The mechanical response of outer hair cells to voltage change is accompanied by a 'gating current', which is manifested as nonlinear capacitance. We also demonstrate this nonlinear capacitance in transfected kidney cells. We conclude that prestin is the motor protein of the cochlear outer hair cell. PMID- 10821265 TI - Proximate humid and dry regions in Jupiter's atmosphere indicate complex local meteorology. AB - Models of Jupiter's formation and structure predict that its atmosphere is enriched in oxygen, relative to the Sun, and that consequently water clouds should be present globally near the 5-bar pressure level. Past attempts to confirm these predictions have led to contradictory results; in particular, the Galileo probe revealed a very dry atmosphere at the entry site, with no significant clouds at depths exceeding the 2-bar level. Although the entry site was known to be relatively cloud-free, the contrast between the observed local dryness and the expected global wetness was surprising. Here we analyse near infrared (around 5 microm) observations of Jupiter, a spectral region that can reveal the water vapour abundance and vertical cloud structure in the troposphere. We find that humid and extremely dry regions exist in close proximity, and that some humid regions are spatially correlated with bright convective clouds extending from the deep water clouds to the visible atmosphere. PMID- 10821266 TI - Closing the spin gap in the Kondo insulator Ce3Bi4Pt3 at high magnetic fields AB - Kondo insulator materials--such as CeRhAs, CeRhSb, YbB12, Ce3Bi4Pt3 and SmB6--are 3d, 4f and 5f intermetallic compounds that have attracted considerable interest in recent years. At high temperatures, they behave like metals. But as temperature is reduced, an energy gap opens in the conduction band at the Fermi energy and the materials become insulating. This contrasts with other f-electron compounds, which are metallic at all temperatures. The formation of the gap in Kondo insulators has been proposed to be a consequence of hybridization between the conduction band and the f-electron levels, giving a 'spin' gap. If this is indeed the case, metallic behaviour should be recovered when the gap is closed by changing external parameters, such as magnetic field or pressure. Some experimental evidence suggests that the gap can be closed in SmB6 (refs 5, 8) and YbB12 (ref. 9). Here we present specific-heat measurements of Ce3Bi4Pt3 in d.c. and pulsed magnetic fields up to 60 tesla. Numerical results and the analysis of our data using the Coqblin-Schrieffer model demonstrate unambiguously a field induced insulator-to-metal transition. PMID- 10821267 TI - Relaxation in polymer electrolytes on the nanosecond timescale AB - The relation between mechanical and electrical relaxation in polymer/lithium-salt complexes is a fascinating and still unresolved problem in condensed-matter physics, yet has an important bearing on the viability of such materials for use as electrolytes in lithium batteries. At room temperature, these materials are biphasic: they consist of both fluid amorphous regions and salt-enriched crystalline regions. Ionic conduction is known to occur predominantly in the amorphous fluid regions. Although the conduction mechanisms are not yet fully understood, it is widely accepted that lithium ions, coordinated with groups of ether oxygen atoms on single or perhaps double polymer chains, move through re coordination with other oxygen-bearing groups. The formation and disruption of these coordination bonds must be accompanied by strong relaxation of the local chain structure. Here we probe the relaxation on a nanosecond timescale using quasielastic neutron scattering, and we show that at least two processes are involved: a slow process with a translational character and one or two fast processes with a rotational character. Whereas the former reflects the slowing down of the translational relaxation commonly observed in polyethylene oxide and other polymer melts, the latter appears to be unique to the polymer electrolytes and has not (to our knowledge) been observed before. A clear picture emerges of the lithium cations forming crosslinks between chain segments and thereby profoundly altering the dynamics of the polymer network. PMID- 10821268 TI - Non-fluorous polymers with very high solubility in supercritical CO2 down to low pressures AB - Liquid and supercritical carbon dioxide have attracted much interest as environmentally benign solvents, but their practical use has been limited by the need for high CO2 pressures to dissolve even small amounts of polar, amphiphilic, organometallic, or high-molecular-mass compounds. So-called 'CO2-philes' efficiently transport insoluble or poorly soluble materials into CO2 solvent, resulting in the development of a broad range of CO2-based processes, including homogeneous and heterogeneous polymerization, extraction of proteins and metals, and homogeneous catalysis. But as the most effective CO2-philes are expensive fluorocarbons, such as poly(perfluoroether), the commercialization of otherwise promising CO2-based processes has met with only limited success. Here we show that copolymers can act as efficient, non-fluorous CO2-philes if their constituent monomers are chosen to optimize the balance between the enthalpy and entropy of solute-copolymer and copolymer-copolymer interactions. Guided by heuristic rules regarding these interactions, we have used inexpensive propylene and CO2 to synthesize a series of poly(ether-carbonate) copolymers that readily dissolve in CO2 at low pressures. Even though non-fluorous polymers are generally assumed to be CO2-phobic, we expect that our design principles can be used to create a wide range of non-fluorous CO2-philes from low-cost raw materials, thus rendering a variety of CO2-based processes economically favourable, particularly in cases where recycling of CO2-philes is difficult. PMID- 10821269 TI - Resolving the 'opal paradox' in the Southern Ocean AB - In the Southern Ocean, high accumulation rates of opal--which forms by precipitation from silica-bearing solutions--have been found in the sediment in spite of low production rates of biogenic silica and carbon in the overlying surface waters. This so-called 'opal paradox' is generally attributed to a higher efficiency of opal preservation in the Southern Ocean than elsewhere. Here we report biogenic silica production rates, opal rain rates in the water column and opal sediment burial rates for the Indian Ocean sector of the Southern Ocean, which show that the assumed opal paradox is a result of underestimated opal production rates and overestimated opal accumulation rates. Our data thus demonstrate that the overall preservation efficiency of biogenic opal in this region is substantially lower than previously thought, and that it lies within a factor of two of the global mean. The comparison of our revised opal preservation efficiencies for the Southern Ocean with existing values from the equatorial Pacific Ocean and the North Atlantic Ocean shows that spatial differences in preservation efficiencies are not the primary reason for the differences in sedimentary opal accumulation. The reconciliation of surface production rates and sedimentary accumulation rates may enable the use of biogenic opal in the reconstruction of palaeo-productivity when the factors that affect the Si/C ratio are better understood. PMID- 10821270 TI - Constraints on the composition of the Earth's core from ab initio calculations AB - Knowledge of the composition of the Earth's core is important for understanding its melting point and therefore the temperature at the inner-core boundary and the temperature profile of the core and mantle. In addition, the partitioning of light elements between solid and liquid, as the outer core freezes at the inner core boundary, is believed to drive compositional convection, which in turn generates the Earth's magnetic field. It is generally accepted that the liquid outer core and the solid inner core consist mainly of iron. The outer core, however, is also thought to contain a significant fraction of light elements, because its density--as deduced from seismological data and other measurements- is 6-10 per cent less than that estimated for pure liquid iron. Similar evidence indicates a smaller but still appreciable fraction of light elements in the inner core. The leading candidates for the light elements present in the core are sulphur, oxygen and silicon. Here we obtain a constraint on core composition derived from ab initio calculation of the chemical potentials of light elements dissolved in solid and liquid iron. We present results for the case of sulphur, which provide strong evidence against the proposal that the outer core is close to being a binary iron-sulphur mixture. PMID- 10821271 TI - Detection and classification of atmospheric methane oxidizing bacteria in soil. AB - Well-drained non-agricultural soils mediate the oxidation of methane directly from the atmosphere, contributing 5 to 10% towards the global methane sink. Studies of methane oxidation kinetics in soil infer the activity of two methanotrophic populations: one that is only active at high methane concentrations (low affinity) and another that tolerates atmospheric levels of methane (high affinity). The activity of the latter has not been demonstrated by cultured laboratory strains of methanotrophs, leaving the microbiology of methane oxidation at atmospheric concentrations unclear. Here we describe a new pulse chase experiment using long-term enrichment with 12CH4 followed by short-term exposure to 13CH4 to isotopically label methanotrophs in a soil from a temperate forest. Analysis of labelled phospholipid fatty acids (PLFAs) provided unambiguous evidence of methane assimilation at true atmospheric concentrations (1.8-3.6 p.p.m.v.). High proportions of 13C-labelled C18 fatty acids and the co occurrence of a labelled, branched C17 fatty acid indicated that a new methanotroph, similar at the PLFA level to known type II methanotrophs, was the predominant soil micro-organism responsible for atmospheric methane oxidation. PMID- 10821272 TI - Infectious parthenogenesis. AB - Parthenogenesis-inducing Wolbachia bacteria are reproductive parasites that cause infected female wasps to produce daughters without mating. This manipulation of the host's reproduction enhances the transmission of Wolbachia to future generations because the bacteria are passed on vertically only from mothers to daughters. Males are dead ends for cytoplasmically inherited bacteria: they do not pass them on to their offspring. Vertical transmission of Wolbachia has been previously considered to be the main mode of transmission. Here we report frequent horizontal transmission from infected to uninfected wasp larvae sharing a common food source. The transferred Wolbachia are then vertically transmitted to the new host's offspring. This natural and unexpectedly frequent horizontal transfer of parthenogensis-inducing Wolbachia intraspecifically has important implications for the co-evolution of Wolbachia and their host. PMID- 10821273 TI - Rewarding effects of opiates are absent in mice lacking the receptor for substance P. AB - Modulation of substance P activity offers a radical new approach to the management of depression, anxiety and stress. The substance P receptor is highly expressed in areas of the brain that are implicated in these behaviours, but also in other areas such as the nucleus accumbens which mediate the motivational properties of both natural rewards such as food and of drugs of abuse such as opiates. Here we show a loss of the rewarding properties of morphine in mice with a genetic disruption of the substance P receptor. The loss was specific to morphine, as both groups of mice responded when cocaine or food were used as rewards. The physical response to opiate withdrawal was also reduced in substance P receptor knockout mice. We conclude that substance P has an important and specific role in mediating the motivational aspects of opiates and may represent a new pharmacological route for the control of drug abuse. PMID- 10821274 TI - Vanilloid receptor-1 is essential for inflammatory thermal hyperalgesia. AB - The vanilloid receptor-1 (VR1) is a ligand-gated, non-selective cation channel expressed predominantly by sensory neurons. VR1 responds to noxious stimuli including capsaicin, the pungent component of chilli peppers, heat and extracellular acidification, and it is able to integrate simultaneous exposure to these stimuli. These findings and research linking capsaicin with nociceptive behaviours (that is, responses to painful stimuli in animals have led to VR1 being considered as important for pain sensation. Here we have disrupted the mouse VR1 gene using standard gene targeting techniques. Small diameter dorsal root ganglion neurons isolated from VR1-null mice lacked many of the capsaicin-, acid- and heat-gated responses that have been previously well characterized in small diameter dorsal root ganglion neurons from various species. Furthermore, although the VR1-null mice appeared normal in a wide range of behavioural tests, including responses to acute noxious thermal stimuli, their ability to develop carrageenan-induced thermal hyperalgesia was completely absent. We conclude that VR1 is required for inflammatory sensitization to noxious thermal stimuli but also that alternative mechanisms are sufficient for normal sensation of noxious heat. PMID- 10821275 TI - Glutamatergic synapses on oligodendrocyte precursor cells in the hippocampus. AB - Fast excitatory neurotransmission in the central nervous system occurs at specialized synaptic junctions between neurons, where a high concentration of glutamate directly activates receptor channels. Low-affinity AMPA (alpha-amino-3 hydroxy-5-methyl isoxazole propionic acid) and kainate glutamate receptors are also expressed by some glial cells, including oligodendrocyte precursor cells (OPCs). However, the conditions that result in activation of glutamate receptors on these non-neuronal cells are not known. Here we report that stimulation of excitatory axons in the hippocampus elicits inward currents in OPCs that are mediated by AMPA receptors. The quantal nature of these responses and their rapid kinetics indicate that they are produced by the exocytosis of vesicles filled with glutamate directly opposite these receptors. Some of these AMPA receptors are permeable to calcium ions, providing a link between axonal activity and internal calcium levels in OPCs. Electron microscopic analysis revealed that vesicle-filled axon terminals make synaptic junctions with the processes of OPCs in both the young and adult hippocampus. These results demonstrate the existence of a rapid signalling pathway from pyramidal neurons to OPCs in the mammalian hippocampus that is mediated by excitatory, glutamatergic synapses. PMID- 10821276 TI - Fringe forms a complex with Notch. AB - The Fringe protein of Drosophila and its vertebrate homologues function in boundary determination during pattern formation. Fringe has been proposed to inhibit Serrate-Notch signalling but to potentiate Delta-Notch signalling. Here we show that Fringe and Notch form a complex through both the Lin-Notch repeats and the epidermal growth factor repeats 22-36 (EGF22-36) of Notch when they are co-expressed. The Abruptex59b (Ax59b) and AxM1 mutations, which are caused by missense mutations in EGF repeats 24 and 25, respectively, abolish the Fringe Notch interaction through EGF22-36, whereas the l(1)N(B) mutation in the third Lin-Notch repeat of Notch abolishes the interaction through Lin-Notch repeats. Ax mutations also greatly affect the Notch response to ectopic Fringe in vivo. Results from in vitro protein mixing experiments and subcellular colocalization experiments indicate that the Fringe-Notch complex may form before their secretion. These findings explain how Fringe acts cell-autonomously to modulate the ligand preference of Notch and why the Fringe-Notch relationship is conserved between phyla and in the development of very diverse structures. PMID- 10821277 TI - ATF-2 has intrinsic histone acetyltransferase activity which is modulated by phosphorylation. AB - Transcription factors carry functional domains, which are often physically distinct, for sequence-specific DNA binding, transcriptional activation and regulatory functions. The transcription factor ATF-2 is a DNA-binding protein that binds to cyclic AMP-response elements (CREs), forms a homodimer or heterodimer with c-Jun, and stimulates CRE-dependent transcription. Here we report that ATF-2 is a histone acetyltransferase (HAT), which specifically acetylates histones H2B and H4 in vitro. Motif A, which is located in the HAT domain, is responsible for the stimulation of CRE-dependent transcription; moreover, in response to ultraviolet irradiation, phosphorylation of ATF-2 is accompanied by enhanced HAT activity of ATF-2 and CRE-dependent transcription. These results indicate that phosphorylation of ATF-2 controls its intrinsic HAT activity and its action on CRE-dependent transcription. ATF-2 may represent a new class of sequence-specific factors, which are able to activate transcription by direct effects on chromatin components. PMID- 10821278 TI - B and C floral organ identity functions require SEPALLATA MADS-box genes. AB - Abnormal flowers have been recognized for thousands of years, but only in the past decade have the mysteries of flower development begun to unfold. Among these mysteries is the differentiation of four distinct organ types (sepals, petals, stamens and carpels), each of which may be a modified leaf. A landmark accomplishment in plant developmental biology is the ABC model of flower organ identity. This simple model provides a conceptual framework for explaining how the individual and combined activities of the ABC genes produce the four organ types of the typical eudicot flower. Here we show that the activities of the B and C organ-identity genes require the activities of three closely related and functionally redundant MADS-box genes, SEPALLATA1/2/3 (SEP1/2/3). Triple mutant Arabidopsis plants lacking the activity of all three SEP genes produce flowers in which all organs develop as sepals. Thus SEP1/2/3 are a class of organ-identity genes that is required for development of petals, stamens and carpels. PMID- 10821279 TI - Disruption of the plant gene MOM releases transcriptional silencing of methylated genes. AB - Epigenetic modifications change transcription patterns in multicellular organisms to achieve tissue-specific gene expression and inactivate alien DNA such as transposons or transgenes. In plants and animals, DNA methylation is involved in heritability and flexibility of epigenetic states, although its function is far from clear. We have isolated an Arabidopsis gene, MOM, whose product is required for the maintenance of transcriptional gene silencing. Mutation of this gene or depletion of its transcript by expression of antisense RNA reactivates transcription from several previously silent, heavily methylated loci. Despite this, the dense methylation at these reactivated loci is maintained even after nine generations, indicating that transcriptional activity and methylation pattern are inherited independently. The predicted MOM gene product is a nuclear protein of 2,001 amino acids containing a region similar to part of the ATPase region of the SWI2/SNF2 family, members of which are involved in chromatin remodelling. MOM is the first known molecular component that is essential for transcriptional gene silencing and does not affect methylation pattern. Thus, it may act downstream of methylation in epigenetic regulation, or be part of a new pathway that does not require methylation marks. PMID- 10821280 TI - Causes, consequences and ethics of biodiversity. AB - The existence of so great a diversity of species on Earth remains a mystery, the solution to which may also explain why and how biodiversity influences the functioning of ecosystems. The answer may lie in quantifying the trade-offs that organisms face in dealing with the constraints of their environment. Societal responses to the loss of biodiversity also involve trade-offs, and the elaboration of these will be essential in developing wiser environmental ethics and policy. PMID- 10821281 TI - Getting the measure of biodiversity. AB - The term 'biodiversity' is a simple contraction of 'biological diversity', and at first sight the concept is simple too: biodiversity is the sum total of all biotic variation from the level of genes to ecosystems. The challenge comes in measuring such a broad concept in ways that are useful. We show that, although biodiversity can never be fully captured by a single number, study of particular facets has led to rapid, exciting and sometimes alarming discoveries. Phylogenetic and temporal analyses are shedding light on the ecological and evolutionary processes that have shaped current biodiversity. There is no doubt that humans are now destroying this diversity at an alarming rate. A vital question now being tackled is how badly this loss affects ecosystem functioning. Although current research efforts are impressive, they are tiny in comparison to the amount of unknown diversity and the urgency and importance of the task. PMID- 10821282 TI - Global patterns in biodiversity. AB - To a first approximation, the distribution of biodiversity across the Earth can be described in terms of a relatively small number of broad-scale spatial patterns. Although these patterns are increasingly well documented, understanding why they exist constitutes one of the most significant intellectual challenges to ecologists and biogeographers. Theory is, however, developing rapidly, improving in its internal consistency, and more readily subjected to empirical challenge. PMID- 10821283 TI - The diversity-stability debate. AB - There exists little doubt that the Earth's biodiversity is declining. The Nature Conservancy, for example, has documented that one-third of the plant and animal species in the United States are now at risk of extinction. The problem is a monumental one, and forces us to consider in depth how we expect ecosystems, which ultimately are our life-support systems, to respond to reductions in diversity. This issue--commonly referred to as the diversity-stability debate--is the subject of this review, which synthesizes historical ideas with recent advances. Both theory and empirical evidence agree that we should expect declines in diversity to accelerate the simplification of ecological communities. PMID- 10821284 TI - Consequences of changing biodiversity. AB - Human alteration of the global environment has triggered the sixth major extinction event in the history of life and caused widespread changes in the global distribution of organisms. These changes in biodiversity alter ecosystem processes and change the resilience of ecosystems to environmental change. This has profound consequences for services that humans derive from ecosystems. The large ecological and societal consequences of changing biodiversity should be minimized to preserve options for future solutions to global environmental problems. PMID- 10821285 TI - Systematic conservation planning. AB - The realization of conservation goals requires strategies for managing whole landscapes including areas allocated to both production and protection. Reserves alone are not adequate for nature conservation but they are the cornerstone on which regional strategies are built. Reserves have two main roles. They should sample or represent the biodiversity of each region and they should separate this biodiversity from processes that threaten its persistence. Existing reserve systems throughout the world contain a biased sample of biodiversity, usually that of remote places and other areas that are unsuitable for commercial activities. A more systematic approach to locating and designing reserves has been evolving and this approach will need to be implemented if a large proportion of today's biodiversity is to exist in a future of increasing numbers of people and their demands on natural resources. PMID- 10821286 TI - Center for Applied Biodiversity Science. Integrating science and conservation. PMID- 10821288 TI - Are mega-mergers good medicine for the pharmaceutical industry? PMID- 10821287 TI - Conservation International and biodiversity conservation. PMID- 10821289 TI - Partners resolve their differences and unite at the second attempt. PMID- 10821290 TI - The Charcot foot. AB - AIMS: To review the clinical manifestations of the Charcot foot in diabetes mellitus, with particular reference to theories concerning aetiology. METHODS: Systematic review of the published literature, searching for the keywords 'Charcot', 'foot and diabetes' and 'neuropathy' on Medline, as well as by examination of the references in recent published reviews. CONCLUSIONS: The Charcot foot of diabetes mellitus is a common problem, and yet is not widely recognized by non-specialists. The failure of professionals to identify the condition in its early phases is probably largely responsible for the gross deformity which follows continued weight-bearing. The condition is confined to those with severe peripheral neuropathy. It is thought to result from three factors: motor neuropathy leading to the development of abnormal forces within the foot, subsequent disorganization of the foot as a result of associated osteopenia and progressive destruction from continued weight-bearing, enabled by reduced pain sensation. The cause of the osteopenia is not known, but it is associated with increased bone blood flow, which may be mainly the result of loss of sympathetic innervation. The importance of increased limb blood flow in the pathogenesis of the Charcot foot has been recognized for over a century. Paradoxically, the increased flow is associated with evidence of macrovascular disease, in that the prevalence of vascular calcification of pedal vessels approaches 90%. After an interval of many months, the condition tends to evolve: the increased blood flow lessens, the osteopenia is reduced and the disorganized bones become sclerotic. This tendency for the condition to evolve remains unexplained, since it would not be expected if the condition was caused solely by progressive denervation. As a result, it is suggested that another factor may be involved in the pathogenesis of the Charcot foot: an abnormal vasomotor reflex, analogous to reflex sympathetic dystrophy, occurring against a background of severe peripheral neuropathy. The resolution of the condition occurs because it is the reflex component of the hyperaemia which proves self-limiting. PMID- 10821291 TI - Sorbitol and myo-inositol levels and morphology of sural nerve in relation to peripheral nerve function and clinical neuropathy in men with diabetic, impaired, and normal glucose tolerance. AB - AIMS: Sorbitol and myo-inositol levels and morphology of sural nerve were compared with nerve function and clinical neuropathy in men with diabetic, impaired (IGT), and normal glucose tolerance. METHODS: After neurography of sural nerve and determinations of sensory thresholds for vibration, warm and cold on the foot, whole nerve sural nerve biopsy was performed in 10 men with Type 1 diabetes mellitus, 10 with IGT, and 10 with normal glucose tolerance. Polyol levels were assessed by gas-liquid chromatography/mass spectrometry. RESULTS: Sural nerve amplitudes were significantly lower and sorbitol levels significantly higher in diabetic patients (median (interquartile range)) (3.7 (3.5) microV and 643 (412) pmol/mg protein, respectively) both compared with IGT (11.3 (10.6)microV; P = 0.04 and 286 (83) pmol/mg protein; P = 0.0032, respectively) and normally glucose tolerant (10.0 (11.6); P = 0.0142 and 296 (250) pmol/mg protein; P = 0.0191, respectively) subjects. There were no differences in nerve morphology between the three groups. Nerve myo-inositol levels correlated, however, positively with cluster density (rs = 0.56; P = 0.0054). In diabetic and IGT subjects, sural nerve amplitudes (2.6 (3.8) vs. 12.1 (10.6) microV; P = 0.0246) and myelinated nerve fibre density (MNFD; 4,076 (1091) vs. 5,219 (668) nerve fibres/mm2; P = 0.0021) were significantly lower in nine subjects with clinical neuropathy than in 10 without. CONCLUSIONS: Nerve degeneration (i.e. MNFD) correlated with clinical neuropathy but not with glucose tolerance status whereas nerve myo-inositol levels positively correlated with signs of nerve regeneration (i.e. increased cluster density). PMID- 10821293 TI - Role of islet autoimmunity in the aetiology of different clinical subtypes of diabetes mellitus in young north Indians. AB - AIMS: To determine the role of islet autoimmunity in the aetiology of different clinical subtypes of diabetes mellitus in young north Indian patients by measuring islet autoantibodies. METHODS: In a cross-sectional study, 145 young patients with diabetes (onset < 30 years) were subdivided into the following categories: Type 1 diabetes (n = 83), malnutrition-modulated diabetes mellitus (MMDM, n = 31) and fibro-calculous pancreatic diabetes (FCPD, n = 31). MMDM subjects presented with emaciation and severe insulin-requiring but ketosis resistant diabetes, while FCPD was associated with idiopathic chronic calcific pancreatitis. Antibodies to glutamic acid decarboxylase (GADA) and IA-2 (IA-2 A) were detected by immunoprecipitation of 35S-labelled recombinant antigens and cytoplasmic islet cell antibody (ICA) by indirect immunofluorescence. RESULTS: GADA were present in a significant proportion (23%) of patients with MMDM. In contrast, IA-2 A was increased only among patients with Type 1 diabetes (22%), but not MMDM (3%, P < 0.05). Among patients with a duration of diabetes < 2 years, GADA and/or IA-2 A were found in 61% of Type 1 diabetic and 37% of MMDM patients (P < 0.01). MMDM patients who were positive for GADA had a shorter duration of diabetes, but did not differ in their age at onset of diabetes, body mass index, fasting plasma C-peptide, or frequency of thyroid microsomal and parietal cell antibodies. FCPD subjects had the lowest prevalence of autoantibodies: IA-2 and ICA were absent, while GADA were present in 7% (P < 0.05 vs. Type 1 diabetes). CONCLUSIONS: GADA, though not IA-2 A, were present in a substantial proportion of patients with the MMDM variant of diabetes, suggesting that islet autoimmunity may play a role in its pathogenesis. In contrast, none of the islet antibodies was increased in subjects with FCPD, making it likely that it is a secondary type of diabetes. PMID- 10821292 TI - Ketoacidosis in young adults is not related to the islet antibodies at the diagnosis of Type 1 diabetes mellitus--a nationwide study. AB - AIMS: To test the hypothesis that there is lower prevalence of islet antibodies in subjects with newly diagnosed Type 1 diabetes mellitus in young adulthood than in children is associated with less severe diabetes at time of diagnosis. METHODS: This investigation was based on a nationwide study (Diabetes Incidence Study in Sweden) of 15-34-year-old newly diagnosed diabetic subjects. During 1992 1993, all diabetic subjects (excluding secondary and gestational diabetes) were reported on standardized forms, with information about clinical characteristics at diagnosis. The study examined islet cell antibodies (ICA) by indirect immunofluorescence, and autoantibodies to glutamic acid decarboxylase (GADA), tyrosine phosphatase-like antigen (IA-2A) and insulin (IAA) as well as C-peptide by radioimmunoassay. RESULTS: Blood samples were available from 78 patients with diabetic ketoacidosis (DKA) and 517 non-acidotic patients. The prevalence of ICA (63% vs. 57%), GADA (63% vs. 66%), IA-2A (35% vs. 44%) and IAA (20% vs. 15%) were very similar in patients with or without DKA. The median levels of the four autoantibodies did not differ between the two groups. High blood glucose (P < 0.001) and low C-peptide levels (P < 0.001) were the only parameters found to be related to DKA. CONCLUSIONS: The similarities in findings of newly diagnosed diabetic patients with or without DKA regarding ICA, GADA, IA-2A and IAA suggest that there is no relationship between the expression of antigenicity and the severity of beta-cell dysfunction. The lower prevalence of the four autoantibodies in 15-34-year-old diabetic subjects compared with previous findings in children is not explained by misclassification of diabetes type. PMID- 10821294 TI - Maternal and perinatal outcomes in 143 Danish women with gestational diabetes mellitus and 143 controls with a similar risk profile. AB - AIMS: To assess maternal and fetal outcomes in pregnancies complicated by gestational diabetes mellitus (GDM) compared to non-diabetic pregnancies with an otherwise similar risk profile and to study the association between different anti-diabetic treatments and fetal outcomes. METHODS: The records of 143 consecutive GDM pregnancies and 143 non-diabetic controls matched on the basis of age, parity and pre-pregnancy body mass index (BMI) were studied. The GDM patients were treated with diet, tolbutamide and insulin. Data were collected from medical records and birth records. RESULTS: Despite treatment, the GDM group had a statistically significant higher frequency of maternal hypertension (20% vs. 11%), induction of labour (61% vs. 24%), Caesarean section (33% vs. 21%), macrosomia (14% vs. 6%), neonatal hypoglycaemia (24% vs. 0) and admission to a neonatal unit (46% vs. 12%). The risk of complications was similar in the different treatment groups. However, in the tolbutamide-treated group, one case of long-standing severe hypoglycaemia in a premature neonate occurred. CONCLUSIONS: Pregnancies complicated by GDM are associated with a higher frequency of adverse maternal and fetal outcomes. The outcomes seem to be unaffected by treatment modality. However, because of the potential risk of hypoglycaemia in some neonates, tolbutamide treatment cannot be recommended in pregnancy. PMID- 10821295 TI - Rosiglitazone taken once daily provides effective glycaemic control in patients with Type 2 diabetes mellitus. AB - AIMS: To evaluate the clinical efficacy and safety of rosiglitazone as a once daily treatment for Type 2 diabetes mellitus (DM). METHODS: Three hundred and sixty-nine patients with Type 2 DM (mean age 63 years; mean body mass index (BMI) 29.4 kg/m2) were enrolled in a double-blind, parallel group, placebo-controlled, dose-ranging study. Patients were randomly assigned to receive placebo or rosiglitazone at doses of 4, 8, or 12 mg daily for 8 weeks. RESULTS: At 8 weeks, fasting plasma glucose (FPG) decreased significantly in the rosiglitazone 4 mg, 8 mg, and 12 mg groups (-0.9, -2.0 and -1.7 mmol/l; P = 0.0003, < 0.0001, and < 0.0001, respectively) compared with placebo (+0.4 mmol/l). The improvements in FPG were dose ordered for 4 and 8 mg/ day. The 12 mg/day dose produced no additional improvement. There were small decreases in haemoglobin and haematocrit in the rosiglitazone treatment groups. The overall incidence of adverse experiences was similar in all treatment groups, including placebo with no evidence of hypoglycaemia or hepatotoxicity. CONCLUSIONS: Rosiglitazone improves glycaemic control when given once daily to treat Type 2 diabetes mellitus and is well tolerated at doses up to and including 12 mg. PMID- 10821296 TI - Young Chinese adults with new onset of diabetic ketoacidosis--clinical course, autoimmune status and progression of pancreatic beta-cell function. AB - AIMS: To examine the clinical course, autoimmune status and pancreatic beta cell function, over a 2-year period, in young Chinese subjects newly presenting with diabetic ketoacidosis (DKA). METHODS: A prospective study involving 562 out of 27,893 patients who were admitted to the medical ward with a principal diagnosis of diabetes mellitus during the recruitment period of 1 year. RESULTS: Of these 562 patients, 27 were aged less than 35 years and admitted with a diagnosis of DKA and 11 (six males and five females) of these were newly diagnosed. Antibodies to glutamic acid decarboxylase (GAD) were present in five patients. Anti-ICA 512 was not detected in any of the patients. Basal and post-glucagon stimulated plasma C-peptide remained in the insulin-deficient range although showing improvement at 2 years. CONCLUSIONS: These findings confirm the relative rarity of autoimmune Type 1 diabetes in young Chinese. Even when the clinical presentation takes the extreme form of acute DKA, less than 50% have positive autoimmune markers. PMID- 10821297 TI - Relation between insulin resistance and carotid intima-media thickness and stenosis in non-diabetic subjects. Results from a cross-sectional study in Malmo, Sweden. AB - AIMS: To assess whether there is an association between insulin resistance and carotid intima-media thickness and stenosis in non-diabetic subjects free from symptomatic cardiovascular disease. METHODS: A cross-sectional population-based study in Malmo, Sweden, of 4,816 (40% men) subjects, born 1926-1945. The prevalence of insulin resistance was established by the homeostasis model assessment (HOMA) and defined as values above the 75th percentile. Criteria issued by the European Group for the Study of Insulin Resistance (EGIR) were used for the definition of the insulin resistance syndrome. Common carotid artery intima-media thickness (IMT) and carotid stenosis (> 15%) were measured by B-mode ultrasonography. RESULTS: Age and sex-adjusted common carotid IMT among subjects with the insulin resistance syndrome (12.7%) and controls was 0.812 mm, respectively, 0.778 mm (P < 0.001). The prevalence of stenosis in the two groups was 22.9 and 19.2% (P = 0.040). Insulin resistance per se was after adjustment for age and sex associated with increased IMT (0.780 mm vs. 0.754 mm, P < 0.001). This association disappeared, however, when other factors included in the insulin resistance syndrome were taken into account. CONCLUSIONS: Fasting serum insulin covaries with a number of factors and conditions known to influence the development of atherosclerosis. It is concluded that the association between insulin resistance, as assessed by the HOMA method in non-diabetic subjects, and atherosclerosis is explained by its covariance with established risk factors for cardiovascular disease of which hypertension seems to be the most significant. PMID- 10821298 TI - Lipoproteins and low-dose estradiol replacement therapy in post-menopausal Type 2 diabetic patients: the effect of addition of norethisterone acetate. AB - AIMS: Low-dose continuous oestrogen/progestogen may increase patient compliance long-term but the cardioprotective effects in diabetes are unknown. The aim of this study was to compare the effect of low-dose oral oestrogen (1 mg, 17-beta estradiol) treatment with oestrogen (1 mg 17-beta-estradiol) in combination with low-dose (0.5 mg) continuous norethisterone acetate (NETA) on lipoproteins in Type 2 diabetic patients. METHODS: Thirty-four post-menopausal Type 2 diabetic patients in moderate control (mean haemoglobin A1c 7.7%) who had a serum oestradiol level of < 50 pg/ml were examined over a 6-month period. Serum lipids, and lipoprotein composition of very low density lipoprotein (VLDL), low density lipoprotein (LDL) and high density lipoprotein (HDL) were measured. Serum lipoprotein(a) was determined by an ELISA method, LDL fatty acids by gas-liquid chromatography and LDL oxidizability by thiobarbituric acid reactive substances (TBARS assay). Cholesteryl ester transfer protein (CETP), and cell cholesterol were measured. RESULTS: There was a reduction in serum cholesterol on both treatments but no significant difference between treatment groups. LDL cholesterol decreased by 17% in each group. There was a no significant difference between the groups in serum VLDL or HDL cholesterol or serum triglycerides during the study. The change in lipoprotein(a) during the study was not significantly different between the groups. There was no significant difference in 4 h LDL oxidizability between groups. Although CETP increased with time in both groups there was no significant difference in the change between the groups. CONCLUSION: In this small study, the addition of continuous low-dose NETA did not reduce the potentially beneficial effects of low-dose 17-beta-estradiol on the progression of atherosclerosis in diabetes. PMID- 10821299 TI - Driving and insulin--consensus, conflict or confusion? AB - AIMS: All drivers requiring insulin treatment must be able to demonstrate satisfactory diabetic control and recognition of hypoglycaemic symptoms before being allowed to drive a motor vehicle. Clinicians have a duty to discuss fitness to drive with their patients. However, is the advice given consistent and in line with the regulations published by the Driver and Vehicle Licensing Authority (DVLA)? METHODS: Six 'real-life' case scenarios were posted to clinicians (consultant diabetologists, specialist registrars and diabetes specialist nurses) within Wessex, UK. The identical cases were also sent to the DVLA for their comments. RESULTS: Sixty-six doctors (36 consultants) and 70 diabetes specialist nurses were contacted by postal questionnaire of which replies were received from 17 consultants (47%), 17 specialist registrars (57%) and 39 diabetes specialist nurses (56%). Although there was general agreement in cases of hypoglycaemia unawareness, there was disagreement where patients had or were at risk of unstable control albeit for a short time. CONCLUSIONS: Patients treated with insulin may receive conflicting information concerning their ability to drive. PMID- 10821300 TI - Increased left ventricular mass index and nocturnal systolic blood pressure in patients with Type 2 diabetes mellitus and microalbuminuria. AB - AIMS: To compare left ventricular mass (LVM) index and function in patients with Type 2 diabetes mellitus with and without microalbuminuria and to investigate the clinical determinants of left ventricular hypertrophy. METHODS: Echocardiography, electrocardiography and 24-h ambulatory blood pressure monitoring were performed in microalbuminuric (n = 29) and normoalbuminuric (n = 29) patients with Type 2 diabetes and no clinical evidence of heart disease. Groups were individually matched for age, sex and diabetes duration and smoking status. RESULTS: LVM index (62 (34-87) vs. 52 (33-89) g/m2.7, P = 0.04) and LVH prevalence, using two out of three definitions, were greater in patients with microalbuminuria (LVM/height2.7: 72 vs. 59%, P = 0.27, LVM/height: 66 vs. 38%, P = 0.04, LVM/body surface area: 59 vs. 31%, P = 0.03). Night-time systolic blood pressure (126 (99-163) vs. 120 (104 157) mmHg, P = 0.005) and the night/day systolic blood pressure ratio (0.92 (0.08) vs. 0.88 (0.06), P = 0.04) were higher in those with microalbuminuria. Systolic and diastolic function were similar in both groups. Linear regression analyses showed that body mass index (BMI) was significantly related to loge LVM index (R2 = 11.8%, P = 0.005) and a relationship with night/day systolic blood pressure was also suggested (R2 = 4.6%, P = 0.057). CONCLUSIONS: In patients with Type 2 diabetes, LVH is more common and severe in those with microalbuminuria. Its presence may be related to raised night/day systolic blood pressure ratio and is significantly related to BMI. The high prevalence of LVH strengthens the case for echocardiographic screening in Type 2 diabetes to identify high risk patients who might benefit from aggressive cardiovascular risk factor intervention. PMID- 10821301 TI - Macular oedema with associated uveitis and cataract following presentation of Type 1 diabetes mellitus in severe ketoacidosis. AB - We present a case of cystoid macular oedema presenting in a newly diagnosed diabetic teenager. She had developed anterior uveitis prior to diabetes and whether this contributed to the subsequent ocular complications remains speculative. The macular changes resolved spontaneously over 6 months without the use of grid laser photocoagulation. PMID- 10821302 TI - Severe diabetic ketoacidosis. PMID- 10821303 TI - Failure to reduce nicotine addiction in young adults with diabetes. PMID- 10821304 TI - The use of lispro for high sugar content snacks between meals in intensive insulin regimens. PMID- 10821305 TI - Tennis leg. PMID- 10821306 TI - Maximum insufflation capacity: vital capacity and cough flows in neuromuscular disease. AB - OBJECTIVE: To investigate the relationships between vital capacity (VC), maximum insufflation capacity (MIC), and both unassisted and assisted peak cough flows (PCFs). DESIGN: The 108 patients were divided into two groups, those whose MICs were greater than their VCs (group 1) and those whose MICs could not exceed their VCs (MIC = VC, or group 2). RESULTS: The MIC correlated positively with the VC for group 1 patients, but the percent increase in MIC correlated negatively with VC. Both VC and MIC correlated significantly with both unassisted and assisted PCF, respectively. Assisting the cough increased the PCF of 37 patients over a previously defined critical level of 2.7 L/sec. The MIC VC difference and percent increase in MIC also correlated significantly with the difference between unassisted and assisted PCF. Although the group 2 patients did not have true cough flows because of inability to close the glottis, their peak expiratory flows were significantly less than the unassisted and assisted PCF of the group 1 patients. CONCLUSIONS: The greater the MIC VC difference, the greater the PCF, and, thereby, the ability to expel airway mucus and avert respiratory complications. The lower the VC, the greater the percent increase in MIC and the greater the percent increase in assisted PCF. Maximal insufflations are extremely important to increase PCF for patients with neuromuscular conditions who have VCs of < 1500 ml. PMID- 10821307 TI - Activation of flexor and extensor trunk muscles in hemiparesis. AB - OBJECTIVE: The purpose of this work was to study the bilateral activity of trunk flexor and extensor muscles in post-stroke hemiparetic patients. DESIGN: Criterion variables were degree of temporal synchronization and level of electromyographic activity in pertinent muscle pairs during two symmetrical tasks: (1) from reclined sitting, bringing the trunk forward; (2) during upright sitting, shrugging shoulders, and extending back. The recti abdomini and external oblique muscles were tested as prime movers of the first task, and the lumbar erector spinae and latissimus dorsi muscles were tested in the second task. Electromyographic recordings from these muscles during three repetitions of each exercise were used for analysis. RESULTS: Cross-correlation analysis pointed to higher temporal synchronization between the abdominal muscles than between the back extensor muscles and in axial compared with para-axial muscles. Differences between patients and controls were found only for the erector spinae muscle pair, which indicates less synchronous activity between the two sides in the patients. The average electromyographic activity level was comparable for corresponding abdominal muscles, both in the patients and in the control subjects. Regarding the back extensor, side differences were detected for the latissimus dorsi muscles in both groups, with the paretic and right side less active in the patients and controls, respectively. CONCLUSIONS: Altogether, the findings do not support the claim of unilateral deficits in the function of trunk muscles in post stroke hemiparetic patients. They emphasize the need for further characterization of the impairment of trunk extensor and flexor muscles in post-stroke patients. PMID- 10821308 TI - The value of trauma scores: predicting discharge after traumatic brain injury. AB - OBJECTIVE: To determine the association of acute variables with disposition after acute hospitalization. DESIGN: Revised Trauma Score (RTS), Injury Severity Score (ISS), and the Combined Trauma Score Injury Severity Score (TRISS(RTS)) were compared with discharge disposition after acute hospitalization of 378 consecutive patients who sustained a traumatic brain injury (TBI) and were treated at a level 1 trauma center between September 1997 and May 1998. RESULTS: Logistic regression modeling found TRISS(RTS) to predict discharge to home with or without home health assistance or inpatient rehabilitation vs. nursing home placement or death. Subsequent modeling, excluding patients who died or went to nursing homes, identified RTS and ISS as predictors of discharge to home with or without home health vs. inpatient rehabilitation. A sensitivity of 97.78% and 93.91% were achieved with these two models when tested on a population of 4,625 patients with TBI treated during the last 10 yr at the same facility. CONCLUSIONS: The results suggest that RTS, ISS, and TRISS(RTS) are predictors of discharge disposition after acute hospitalization with TBI and may be useful measures of rehabilitation services resource planning early in the course of TBI management. PMID- 10821309 TI - Community Integration Questionnaire for patients with brain tumor: a comparative study. AB - OBJECTIVES: This study compares mean scores obtained on the Community Integration Questionnaire among patients with brain tumor and published data for traumatic brain injury and nonpatient groups. DESIGN: Subjects included 33 adult outpatients with biopsy-confirmed high-grade malignant brain tumors. Both sexes were represented. Subjects were older and better educated than historic samples. The Community Integration Questionnaire was positively skewed. RESULTS: Subjects had higher scores on the Social Integration Scale than on Productivity. Higher education was associated with Productivity and the Community Integration Questionnaire total score. However, greater age was associated with lower scores. Significant gender effects were obtained. CONCLUSIONS: Women with brain tumors had significantly higher Home Integration Scores than males with tumors. However, Home Integration was lower for persons with brain tumors than for either the corresponding gender in the traumatic brain injury model systems group for males and females, respectively. Similarly, compared with community-dwelling persons with traumatic brain injury, males, but not females, with cerebral tumors had significantly lower Social Integration scores. When compared with nondisabled adults, persons with brain tumors reported significantly lower Productivity and Community Integration Questionnaire total scores. PMID- 10821310 TI - Hip hiking and circumduction: quantitative definitions. AB - OBJECTIVE: To define and propose clinically useful quantitative measurements of hip hiking and circumduction using standard three-dimensional motion analysis techniques. DESIGN: We studied pelvic, hip, and thigh motions in 23 subjects with hemiparetic, stiff-legged gait as a result of stroke and compared these motions with those obtained from 23 able-bodied controls. RESULTS: We observed significantly increased hip abduction on the unaffected limb during stance, with simultaneous elevation of the affected side of the pelvis during swing. We define these differences as hip hiking and, thus, can quantify the degree to which hip hiking occurs by measuring the unaffected coronal hip angle and/or the coronal pelvic angle when the affected limb is in midswing. We also observed a greater than normal coronal thigh angle during midswing of the affected limb that we can use to quantitatively define circumduction. Of note, hip abduction during swing was not increased on the affected swing limb, compared with the control. CONCLUSIONS: Hip hiking can be defined precisely as unaffected coronal hip and/or pelvic angle when the affected limb is in midswing and circumduction can be defined as greater than normal coronal thigh angle during midswing of the affected limb. These precise definitions should allow us to better communicate and understand the implications of these gait patterns, and can serve as the basis for clinically meaningful quantitative assessment and outcome measurement tools. PMID- 10821311 TI - Visual testing for readiness to drive after stroke: a multicenter study. AB - OBJECTIVE: The purpose of this study was to determine the ability of a visual perception assessment tool, the Motor-Free Visual Perception Test, to predict on road driving outcome in subjects with stroke. DESIGN: This was a retrospective study of 269 individuals with stroke who completed visual-perception testing and an on-road driving evaluation. Driving evaluators from six evaluation sites in Canada and the United States participated. Visual-perception was assessed using the Motor-Free Visual Perception Test. Scores range from 0 to 36, with a higher score indicating better visual perception. A structured on-road driving evaluation was performed to determine fitness to drive. Based on driving behaviors, a pass or fail outcome was determined by the examiner. RESULTS: The results indicated that, using a score on the Motor-Free Visual Perception Test of < or =30 to indicate poor visual-perception and >30 to indicate good visual perception, the positive predictive value of the Motor-Free Visual Perception Test in identifying those who would fail the on-road test was 60.9% (n = 67/110). The corresponding negative predictive value was 64.2% (n = 102/159). Univariate logistic regression analyses revealed that older age, low Motor-Free Visual Perception Test scores and a right hemisphere lesion contributed significantly to identifying those who failed the on-road test. CONCLUSIONS: The predictive validity of the Motor-Free Visual Perception Test is not sufficiently high to warrant its use as the sole screening tool in identifying those who are unfit to undergo an on-road evaluation. PMID- 10821312 TI - Blood volume and hemoglobin after spinal cord injury. AB - OBJECTIVE: The main purpose of this study was to investigate the difference in total blood volume and hemoglobin mass between spinal cord-injured and able bodied individuals. DESIGN: Total blood volumes of 13 able-bodied and 10 spinal cord-injured individuals (lesion >T4) were determined using the carbon monoxide method. The reproducibility of the total blood volume determination in our setting and the effect of increased physical activity were assessed. RESULTS: Comparison of groups showed a significantly higher hemoglobin mass in able-bodied compared with spinal cord-injured individuals. The total blood volume expressed per kilogram of body mass in able-bodied individuals was significantly greater than in spinal cord-injured individuals. CONCLUSIONS: These results suggest that total blood volume and hemoglobin mass are decreased in spinal cord-injured individuals with a lesion above T4, which may be related to their inactive lifestyle, because total blood volume increased with increased physical activity in these subjects. PMID- 10821313 TI - Anxiety, depressive, or cognitive disorders in rehabilitation patients: effect on length of stay. AB - OBJECTIVE: To test the hypothesis that anxiety, depressive, or cognitive disorders are associated with an increase in length of stay of physical rehabilitation inpatients. DESIGN: Secondary analysis of a 1-yr prospective data recording. Three treatment and rehabilitation centers in the Canton of Vaud (Switzerland). Ninety-five percent of inpatients admitted from November 15, 1990, to November 14, 1991, agreed to participate. Apart from length of stay, data consisted of demographic and medical data results from the Hospital Anxiety and Depression Scale, Mini-Mental State Score, and Functional Autonomy Measurement System. Multivariate linear regression was used in the analysis. RESULTS: The presence of anxiety or depression altered length of stay in a bivariate analysis, although all effects disappeared in a multivariate approach. Factors that had an independent association with length of stay were gender, length of stay in an acute care hospital before hospitalization, treatment and rehabilitative centers, Functional Autonomy Measurement System mobility score, and Functional Autonomy Measurement System Activities of Daily Living score. Results concerning the association between cognition abilities and length were similar. CONCLUSIONS: Our results recognize that an influence of psychiatric disorders acted on length of stay through a relationship between the psychiatric status and the control variables. If mental state influences physical state, then early intervention studies are desirable. If somatic state induces mental alterations, then interventions directed toward the psychiatric sphere will bring mostly qualitative benefits (amelioration of well-being without remarkable effects on length of stay). PMID- 10821314 TI - Functional independence measure prediction: an initial evaluation of residents' skills. AB - The purpose of this preliminary study was to assess the ability of first-year PM&R residents to accurately predict functional outcome in an acute rehabilitation setting. Although statistically significant, the accuracy and precision of the residents' predictive skills were rather modest. A future focus on innovative methods to evaluate and develop these clinical skills appears warranted. PMID- 10821315 TI - Evaluating bone mineral density in osteoporosis. PMID- 10821316 TI - Prescribing exercise training for patients with defibrillators. AB - Patients with an implantable cardioverter defibrillator (ICD) often refrain from physical exercise for fear of precipitating a life-threatening arrhythmia or receiving an ICD shock. However, most of these patients are able to safely exercise if they are provided appropriate clinical guidelines. This review describes the factors that enter into the development of an exercise program for patients with an ICD. PMID- 10821317 TI - Tourniquet paralysis after primary nerve repair. AB - Paralysis after use of a tourniquet during surgery is a well recognized complication. Based on the electrophysiologic findings, 35 cases referred to our electromyography laboratory have been diagnosed as tourniquet paralysis within a 3-yr period. This preliminary report suggests that tourniquet paralysis should be considered during clinical and electrophysiologic evaluations after primary nerve repairs. PMID- 10821318 TI - Structural equation modeling and rehabilitation research. AB - Structural equation modeling (SEM) has been used extensively in the social sciences in the recent past to model human behavior. SEM has been less used by other disciplines, including rehabilitation medicine. This article begins by providing an introduction to structural equation modeling through the discussion of the definition and basic concepts behind SEM using a conceptual model relevant to rehabilitation medicine that describes the pathway from health to disability (the Enabling-Disabling model). The next focus is on several potential pitfalls of which the researcher needs to be aware when using SEM. After this discussion, a hypothetical example is presented using structural equation modeling to evaluate the Enabling-Disabling model. The article concludes with a review of the advantages and disadvantages of using SEM for clinical and social science research. Throughout the article, references are provided for a more detailed examination of SEM. PMID- 10821319 TI - Compensation and a psychiatric model of chronic pain. PMID- 10821320 TI - Comparison of three methods for recording tibial H reflex: a clinical note. PMID- 10821321 TI - Genetic and environmental factors in the aetiology of simple goitre. AB - The aetiology of simple goitre, affecting up to 5% of a population in iodine sufficient areas and over 10% in endemic areas, is incompletely understood. It is generally believed that the development of simple goitre, whether endemic or sporadic, depends on complex interactions between genetic, environmental and endogenous factors. The importance of genetic factors is evident from the clustering of simple goitre within families and from a higher concordance rate for goitre in monozygotic than in dizygotic twins. Recently, studies assessing the role of specific candidate genes or genetic markers in the aetiology of simple goitre have given conflicting data in various families. However, there may well be single genes playing a major role within certain families, eg the thyroglobulin (Tg) gene, the thyroid-stimulating hormone receptor (TSHR) gene, the Na+/I- symporter (NIS) gene, and the multinodular goitre marker 1 (MNG1) on chromosome 14, but the genes will vary from family to family. In addition, family and twin studies also indicate a modest to major role for environmental factors in the aetiology of simple goitre. Clearly, iodine deficiency and cigarette smoking are the most important environmental risk factors associated with the genesis of simple goitre. Other suggested risk factors include naturally occurring goitrogens, emotional stress and certain drugs and infections. Ongoing studies focus on whole-genome screening in multiplex families as well as on large population-based case-control studies. However, the possibility that simple goitre is a heterogeneous disease without a single well-defined genotype and phenotype should be left open. PMID- 10821322 TI - Increasing importance of viruses in acute otitis media. AB - Acute otitis media is generally considered a simple bacterial infection that can be effectively treated with antibiotics. However, despite the extensive use of broad-spectrum antibiotics, poor clinical response to treatment of acute otitis media is common in children. Numerous studies ranging from animal experiments to extensive clinical studies have clearly demonstrated that respiratory viruses play a crucial role in the aetiology and pathogenesis of acute otitis media. Viral infection of the upper respiratory tract initiates the whole cascade of events that finally leads to the development of acute otitis media as a complication. Respiratory viruses induce a release of inflammatory mediators in the nasopharynx, increase bacterial colonization and adherence, and have a suppressive effect on the host's immune defense. Recent data indicate that at least some types of viruses actively invade the middle ear. Viruses also seem to enhance the inflammatory process in the middle ear and impair the outcome of the disease. Vaccines against the major viruses predisposing to acute otitis media hold a great promise for the prevention of this disease. Major advances in the management of acute otitis media will require further research into the mechanisms of viral infection, viral-bacterial interaction and the host inflammatory response during viral infection. PMID- 10821323 TI - Non-lipid-related effects of statins. AB - The beneficial effects of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) on coronary events have generally been attributed to their hypocholesterolaemic properties. However, as mevalonate and other intermediates of cholesterol synthesis (isoprenoids) are necessary for cell proliferation and other important cell functions, effects other than cholesterol reduction may explain the pharmacological properties of statins. In the present review, we discuss the current knowledge on the nonlipid-related effects of statins, with a special emphasis on their potential benefits in different diseases, such as atherosclerosis and cancer. The mechanism(s) responsible for their favourable properties are also reviewed. PMID- 10821324 TI - Preventive treatment for recurrent febrile seizures. AB - Febrile seizures are the most common convulsive events in childhood occurring in 2-5 % of children. 20-30% of these children will have a recurrence during a subsequent febrile infection. Even though the outcome of febrile seizures is benign, the possibility of recurrences keeps most families in fear for years after the first seizure event. Each febrile infection the child experiences increases the risk of recurrence, and there is a positive correlation between the height of the temperature during an infectious disease and the occurrence of febrile seizures. However, prophylactic use of antipyretics does not decrease the recurrence rate. Intermittent use of diazepam during febrile episodes prevents febrile seizures only in selected child populations. The continuous use of antiepileptic drugs is no longer warranted because their side-effects outweigh their benefits. The number of febrile episodes is the only risk factor that can be influenced by preventive measures. The time being, we can reassure parents on the benign nature of febrile seizures. PMID- 10821325 TI - Androgens and the control of skeletal muscle protein synthesis. AB - Athletes have long supported the concept that anabolic steroids increase skeletal muscle mass. However, it was only recently that both testosterone and its synthetic analogue, oxandrolone, were proven capable of inducing myotrophic effects in postabsorptive human skeletal muscle. These findings have provided the physiological evidence that anabolic steroids deserve attention in the clinical arena as a pharmacological intervention against losses in lean body mass associated with age, disease, trauma and burn injury. However, we are lacking in vivo molecular evidence that would directly or indirectly link androgens and the androgen receptor with increases in skeletal muscle mass. Clearly, a need exists to link in vivo and in vitro studies from both the physiological and molecular arena as they relate to androgens and the control and regulation of skeletal muscle mass. In this brief review, newly discovered information and emerging theories relating to the direct, indirect, priming and antiglucocorticoid action of androgens on skeletal muscle will be presented. PMID- 10821326 TI - Impact of serotonin on tumour growth. AB - Several opposite effects of serotonin (5HT) on tumour growth have been reported. On one hand, 5HT is known as a growth factor for several types of nontumoural cells, and it has been proposed to take part in the autocrine loops of growth factors contributing to cell proliferation in aggressive tumours such as small cell lung carcinoma. Depending on the tumour type either 5HT2 or 5HT1 receptor antagonist have been found to inhibit the 5HT-induced increase in tumour growth. In contrast, several authors have also reported that 5HT and 5HT2 agonist can inhibit tumour growth. Most often this effect has been considered to be related with the specific vasoconstrictive effect of 5HT or 5HT2 agonists on the vessels irrigating the tumour, which has been evidenced by intravital microscopy. Intravital microscopy studies have also shown that vessels perfusing the tumour exhibit a specific vasconstrictive response to 5HT1 agonists. In addition, 5HT has been shown to be involved in the effects of several anticancer treatments associated with the reduction of tumour flow. Finally, the specific vasoconstrictive effect of 5HT or 5HT receptor subtype agonists might also be useful in inducing hypoxia in tumours, which could be exploited in a strategy using hypoxia-selective cytotoxins or hypoxia-selective gene therapy. PMID- 10821327 TI - Serotonin and the heart. AB - Serotonin is a naturally occurring vasoactive substance that has diverse cardiophysiological effects. These effects can be explained by the existence of serotonin receptor subtypes which mediate different biological actions. The vasoconstrictive actions of serotonin are mediated by 5-HT2 serotonergic receptors, and serotonin also amplifies the release and activities of other vasoconstrictors, such as angiotensin and norepinephrine. Abnormalities in the serotonergic system may play an important role in the pathophysiology of multiple cardiovascular disease states such as systemic hypertension, primary pulmonary hypertension and peripheral vascular disease. Selective 5-HT2 serotonergic receptor blockers have been developed which appear to be potent vasodilators with therapeutic potential in various cardiovascular disease states. The largest clinical experience has been collected with ketanserin, and other agents in this class are being investigated. Prolongation of the ECG QT interval with 5-HT2 serotonergic receptor blockers may pose a potential risk with these treatments in some patients. PMID- 10821328 TI - Role of serotonin in memory impairment. AB - As a result of its presence in various structures of the central nervous system serotonin (5-HT) plays a role in a great variety of behaviours such as food intake, activity rythms, sexual behaviour and emotional states. Despite this lack of functional specialization, the serotonergic system plays a significant role in learning and memory, in particular by interacting with the cholinergic, glutamatergic, dopaminergic or GABAergic systems. Its action is mediated via specific receptors located in crucial brain structures involved in these functions, primarily the septo-hippocampal complex and the nucleus basalis magnocellularis (NBM)-frontal cortex. Converging evidence suggests that the administration of 5-HT2A/2C or 5-HT4 receptor agonists or 5-HT1A or 5-HT3 and 5 HT1B receptor antagonists prevents memory impairment and facilitates learning in situations involving a high cognitive demand. In contrast, antagonists for 5 HT2A/2C and 5-HT4, or agonists for 5-HT1A or 5-HT3 and 5-HT1B generally have opposite effects. A better understanding of the role played by these and other serotonin receptor subtypes in learning and memory is likely to result from the recent availability of highly specific ligands, such as 5-HT1A, 5-HT1B, 5-HT2A receptor antagonists, and new molecular tools, such as gene knock-out mice, especially inducible mice in which a specific genetic alteration can be restricted both temporally and anatomically. PMID- 10821329 TI - Separate systems for serotonin and leptin in appetite control. AB - Appetite control involves an integration of the drive signals arising form energy stores in the body with the satiety signals generated by periodic episodes of food consumption. Serotonin (5-hydroxytryptamine, 5-HT) has been implicated in the processes of within-meal satiation and postmeal satiety (5-HT1B and 5-HT2C postsynaptic receptors) which are concerned with the signals arising form the pattern of food intake. Central nervous system (CNS) 5-HT is sensitive to circulating levels of the precursor tryptophan, certain macronutrients and peripheral satiety factors such as cholecystokinin (CCK) and enterostatin. Hypothalamic 5-HT receptor systems inhibit neuropeptide Y (NPY), a potent stimulator of hunger and food intake. In contrast to the linking of 5-HT with the consequences of food ingestion, the hormone leptin (OB protein) is regarded as a signal linking adipose tissue status with a number of key CNS circuits. Leptin itself stimulates CNS leptin receptors (OB-r receptor) which link with pro opiomelanocortin (POMC)/ MC-4 receptors. The effects of leptin may also be modulated by factors such as the corticotrophin-releasing factor (CRF), cocaine and amphetamine-regulated transcript (CART), orexins and galanin. Very little evidence exists to support any direct link between the actions of 5-HT and leptin, suggesting that they are separate systems. 5-HT is a part of an integrated network for short-acting satiety signals (episodic in nature), and leptin is a hormonal indicator of long-term (tonic) energy reserves. At a conceptual level, these may represent the distinction between 'satiety' and 'drive'. Interestingly, both 5-HT and leptin modulate the action of NPY, which may form a part of a common output pathway for the expression of appetite. PMID- 10821330 TI - Hypertension and arterial stiffness: the Atherosclerosis Risk in Communities Study. ARIC Investigators. AB - Our objective was to describe the relationship of arterial stiffness and hypertension in a large, population-based sample of men and women. Hypertension related increases in arterial stiffness may reflect the distending pressure and/or structural alterations in the artery. Included were 10,712 participants, ages 45 to 64 years, of the Atherosclerosis Risk in Communities Study, free of prevalent cardiovascular disease. Hypertension was classified as systolic or diastolic blood pressure (BP) > or =140/90 mm Hg, respectively, or the current use of antihypertensive medications. Common carotid arterial diameter change was measured using B-mode ultrasound and an electronic device that utilized radio frequency signals to track the motion of the arterial walls. Using statistical models to control for diastolic BP and pulse pressure, arterial diameter change was calculated separately in normotensive/ nonmedicated and medicated hypertensives. Hypertension was associated with a smaller adjusted diameter change (ie, greater stiffness) in comparison to optimal blood pressure (BP < 120/80 mm Hg): normotensive/nonmedicated men, 0.33 versus 0.43 mm (P < 0.001); medicated men, 0.34 versus 0.42 mm (P < 0.001); normotensive/ nonmedicated women, 0.34 versus 0.40 mm (P < 0.001), and medicated women, 0.33 versus 0.40 mm (P < 0.001). The relationship between pulse pressure and diameter change (ie, the slope of pulse pressure and diameter change) did not differ between hypertensives and normotensives. These cross-sectional data suggest that hypertension is associated with carotid arterial stiffness; however, these differences in the calculated stiffness appear to be the effect of distending pressure rather than structural changes in the carotid artery. PMID- 10821331 TI - Arterial wall thickening at different sites and its association with left ventricular hypertrophy in newly diagnosed essential hypertension. AB - The impact of hypertension on vascular structure at different arterial sites and the relation of vascular hypertrophy with left ventricular (LV) hypertrophy in the early stages of essential hypertension are unclear. In 96 newly diagnosed, never-treated, uncomplicated hypertensive subjects aged < 55 years (43 +/- 9 years, 68 men, clinic blood pressure 152/99 mm Hg, 24-h blood pressure 135/89 mm Hg), we measured LV mass (M-mode echocardiography) and intima-media thickness (IMT) of the carotid and femoral arteries (high-resolution B-mode ultrasound). The average of 24 carotid and 24 femoral IMT readings (common and internal carotid or common and superficial femoral, right and left side, far and near wall, three sampling points per segment) was analyzed. Carotid and femoral IMT were strongly related to each other (r = 0.77). Subjects with LV hypertrophy (n = 33) had a greater IMT at the carotid (0.84 +/- 0.2 v 0.71 +/- 0.2 mm, P < .0001) and femoral (0.77 +/- 0.1 v 0.64 +/- 0.1 mm, P < .0001) level. Carotid IMT showed a positive correlation with LV mass (r = 0.46) and age (r = 0.38), and an inverse one with high-density lipoprotein (HDL) cholesterol (r = -0.26). Femoral IMT was associated positively to LV mass (r = 0.50), age (r = 0.33) and triglycerides (r = 0.29), and inversely to HDL-cholesterol (r = -0.33). The association between IMT (both carotid and femoral) and LV mass held after controlling for age and other confounders in a multiple regression analysis. In summary, in the early stages of hypertension arterial wall thickening appears to be a diffuse process, which occurs in parallel at the carotid and femoral level and shows a positive association with LV hypertrophy. PMID- 10821332 TI - Afferent-efferent vessel dysfunction appears to be a specific characteristic of a large subset of patients with essential hypertension. AB - Exercise renography makes it possible to subdivide essential hypertensives (EHs) into two distinct populations. Fifty to 60% develop exercise-mediated renal dysfunction and a transitory, severe reduction of glomerular filtration. The other subset of EHs does not have exercise-mediated renal dysfunction. We hoped to learn whether the disturbance is also present while EHs rest. Twenty-six EHs and 21 normotensive controls were studied with a resting sequential renogram using Tc-99m-mercaptoacetyl-triglycine (MAG3), a tracer excreted primarily by proximal tubular cells. EHs also had an exercise renogram. All persons had three consecutive 10-min dual-tracer infusion clearance determinations with 111In-DTPA and 131I-hippurate, for the simultaneous determination of glomerular filtration rate (GFR) and effective renal plasma flow (ERPF). To demonstrate the accuracy of the clearance procedure we sought to reproduce Hollenberg's results which show greater flow variability in EH than in normotension. Following this, the variability (VAR) of the GFR and ERPF as well as the variability fraction (VF), the ratio of GFR variability divided into the ERPF variability, were calculated. Hollenberg's results were reproduced. Sixteen of 26 essential hypertensives developed exercise-mediated renal dysfunction. GFR-VAR in EH differed from controls. The VF suggests that EHs with a bilateral abnormal exercise renogram have a more pronounced GFR variability than those EH with a normal exercise renogram. The results point to intraglomerular pressure fluctuations in patients with EH, and the VF suggests that this may be more pronounced in EHs with a bilateral abnormal exercise renogram than in those with a normal exercise renogram. It is hypothesized that the variable GFR provokes renin secretion in EH. PMID- 10821333 TI - Mild left ventricular hypertrophy in essential hypertension: is it really arrhythmogenic? AB - Left ventricular hypertrophy (LVH) has been associated with an increased incidence of ventricular arrhythmias and sudden cardiac death in hypertensive patients. However, it is not known whether this relationship exists in early asymptomatic hypertensives with mild LVH. We prospectively examined 100 consecutive patients with essential hypertension, 35 without and 65 with mild LVH on echocardiography. All underwent a detailed noninvasive arrhythmia work-up and were subsequently followed-up for 3 +/- 1 years in an ambulatory hypertension clinic. None of the 12-lead electrocardiographic parameters examined differed between the two hypertensive groups. A similarly low incidence of simple forms of ventricular ectopy was present in both groups, whereas complex forms of ventricular ectopy were extremely rare in either group. The signal-averaged electrocardiographic parameters examined were also not significantly affected by the presence of mild LVH. Arrhythmia-related symptoms or malignant ventricular arrhythmia events were not observed in either group of patients during follow-up with antihypertensive treatment. The latter resulted in LVH regression in the 65 patients with mild LVH at baseline. It appears that mild LVH among ambulatory hypertensive patients does not carry an additive arrhythmogenic risk and can be successfully reversed with the appropriate antihypertensive therapy, with no need of additional antiarrhythmic management. PMID- 10821334 TI - Heterogeneity of renal cortical circulation in hypertension assessed by dynamic computed tomography. AB - The aim of this study was to assess the grade of heterogeneous disturbance in the renal cortical circulation using dynamic computed tomography and to investigate the relationship between the heterogeneity of renal cortical circulation and hypertension. We studied 125 patients who underwent dynamic computed tomography (CT) for various abdominal diseases and had no serious hemodynamic abnormalities. In dynamic computed tomography under appropriate conditions, each pixel (image element), less than 1 mm2, has a CT number that is in proportion to the concentration of contrast media, which reflects the blood volume in the pixel. The image was constructed at the hilus level about 50 s after the start of a continuous infusion of contrast medium. The mean and standard deviation were calculated from the CT numbers in the renal cortex. The coefficient of variation, ie, the standard deviation divided by the mean value, was used as the index of the heterogeneity of renal cortical circulation. The coefficient of variation was significantly (P < .001) greater in the hypertensive patients (n = 48, 0.174 +/- 0.006 [mean +/- SE]) than in normotensive subjects (n = 77, 0.140 +/- 0.004). The coefficient increased in parallel with the patient's age and with the grade of renal surface irregularity. In the patients whose serum creatinine levels were normal, this parameter also had a significant relationship (r = 0.367, P < .0001) with serum creatinine. These results suggest that the heterogeneity of renal cortical circulation is increased in hypertension and is also associated with aging. This parameter may become a sensitive indicator to detect slight deterioration in the renal cortical circulation. PMID- 10821335 TI - Microalbuminuria in never-treated hypertensives: lack of relationship to hyperinsulinemia and genetic predisposition to hypertension. AB - We evaluated the relationship of microalbuminuria to hyperinsulinemia and family history of hypertension in 92 never-treated essential hypertensives (mean 24-h blood pressure >140 or 90 mm Hg), with positive (F+) or negative (F-) family history of hypertension: 31 had microalbuminuria (MA+) (urinary albumin excretion [UAE], 30 to 300 mg/24 h) and 61 had normal (<30 mg/24 h) UAE (MA-). Glucose and insulin values before and 30, 60, 90, and 120 min after an oral glucose load were measured together with an index of peripheral insulin activity (10(4)/ insulin x glucose values at glucose peak). Subjects with and without microalbuminuria did not differ with regard to age, sex, body mass index, and 24-h heart rate, whereas 24-h, daytime, and nighttime systolic and diastolic blood pressure were significantly higher in MA+ than MA- patients. The prevalence of positive family history of hypertension was similar between MA+ and MA-, as were fasting and stimulated glucose and insulin values and the index of peripheral insulin activity. Subdividing the patients on the basis of family history of hypertension (59 F+, 33 F-) UAE was not significantly different between F+ and F-. UAE did not correlate with glucose and insulin parameters. From our results, in never-treated hypertensives, microalbuminuria is associated with higher blood pressure values, but is related neither to genetic predisposition to hypertension, nor to hyperinsulinemia; therefore, impaired insulin sensitivity and microalbuminuria are two components of the hypertensive syndrome, largely independent of each other. PMID- 10821336 TI - Fibrinolytic function in diuretic-induced volume depletion. AB - Accumulating evidence suggests that the renin-angiotensin system (RAS) may participate in the regulation of fibrinolytic function. In clinical studies, however, angiotensin-converting enzyme (ACE) inhibitors and the angiotensin II receptor antagonist losartan have failed to consistently affect endogenous fibrinolysis. Because such an effect may depend on the degree of prestimulation of the RAS, we have studied parameters of fibrinolytic function in 15 healthy volunteer subjects during baseline (day 1) and after 10 days of treatment with 25 mg of hydrochlorothiazide (HCT)/day (day 11). On the last day of the study (day 12), a single oral dose of 50 mg of losartan was given to the volunteers in addition to HCT and fibrinolytic function was assessed at the peak effect of losartan (5 h later). Plasma renin activity (PRA) was significantly stimulated during diuretic treatment (1.35 +/- 0.21 v 0.34 +/- 0.06 ng mL(-1) x h(-1) [P < .001]) and further increased after losartan (6.39 +/- 1.16 ng mL(-1) x h(-1) [P < .001]). No effects of either the diuretic or losartan could be observed on tissue type plasminogen activator (t-PA) antigen concentration and activity. However, 10 days of treatment with HCT significantly increased plasminogen activator inhibitor-1 (PAI-1) antigen (26.8 +/- 5.8 v 21.1 +/- 3.4 ng/mL [p = .037]). In addition, PAI-1 activity was also tentatively raised by HCT treatment (5.48 +/- 1.82 v 3.88 +/- 0.79 IU/mL [P = .067]). In spite of the marked further rise in PRA after losartan, the stimulation of PAI-1 antigen and activity was blunted by losartan (24.4 +/- 3.6 ng/mL and 4.55 +/- 0.99 IU/mL, respectively). Our results demonstrate that volume depletion induced by HCT treatment is associated with a rise in PAI-1. Acute administration of losartan is capable of blunting this effect, suggesting that the angiotensin II type 1 receptor may participate in this effect of angiotensin II. PMID- 10821338 TI - Pioglitazone attenuates basal and postprandial insulin concentrations and blood pressure in the spontaneously hypertensive rat. AB - Subjects with hypertension are hyperinsulinemic and resistant to insulin stimulated glucose uptake. A similar paradigm is found in the spontaneously hypertensive rat (SHR). These findings suggest the possibility that insulin resistance and hyperinsulinemia may play an important role in blood pressure regulation. Pioglitazone, a thiazolidinedione derivative, sensitizes target tissues to insulin and decreases hyperglycemia and hyperinsulinemia in various insulin-resistant animals. The purpose of this study was to assess the influence of pioglitazone administration on pre- and postprandial glucose and insulin concentrations and determine whether changes in beta-cell secretion resulted in any change in blood pressure measurements. Twelve SHR were fed custom diets ad libitum, six with and six without pioglitazone (20 mg/kg chow). Fasting and postprandial glucose levels were unaltered by pioglitazone treatment. Fasting insulin concentrations were similar at week 1, but were significantly lower (P < .01) in the pioglitazone group at weeks 3 (1.89 +/- 0.3 v7.94 +/- 1.5 ng/mL) and 4 (4.5 +/- 1.4 v9.1 +/- 0.7 ng/mL), compared with the control group. Pioglitazone also significantly (P < .01) lowered postprandial insulin concentrations after an oral glucose challenge. Systolic, mean, and diastolic blood pressures were significantly lower (P < .01), 177 +/- 3 v190 +/- 4.7 mm Hg, 162 +/- 2.1 v175 +/- 5.9 mm Hg, and 156 +/-2.1 v168 +/- 6.2 mm Hg, respectively, in the animals receiving pioglitazone versus the control group. Heart rate, body weight, serum cholesterol, and triglyceride levels were comparable between the two groups. In conclusion, pioglitazone significantly decreased fasting and postprandial insulin concentrations and effectively lowered blood pressure in the SHR. PMID- 10821337 TI - Inhibitors of Na-K-ATPase in human urine: effects of ouabain-like factors and of vanadium-diascorbate on calcium mobilization in rat vascular smooth muscle cells: comparison with the effects of ouabain, angiotensin II, and arginine-vasopressin. AB - Endogenous ouabain-like factors (OLF) may play a role in the pathogenesis of volume-dependent hypertension by raising intracellular free calcium ([Ca2+]i) as a consequence of inhibition of the sodium pump. In previous studies we described the presence of two low molecular (Mr approximately equals 400) inhibitors of Na K-ATPase in human urine, ie, a more polar OLF-1 and a more apolar OLF-2. We subsequently identified the active compound in OLF-2 as vanadium (V(IV)) diascorbate (Mr 416). OLF-1, OLF-2, and V-diascorbate inhibited dose-dependently porcine Na-K-ATPase in vitro. In the present study we investigated the effects of urinary OLF-1, OLF-2, and V-diascorbate on calcium mobilization, ie, on [Ca2+]i in cultured rat vascular smooth muscle (VSM) cells in comparison to the effects of ouabain, angiotensin II (A II), and arginine-vasopressin (AVP). [Ca2+]i was determined by the fura-2 method. OLF-1 and OLF-2 (each approximately equals 10( 4) mol/L), obtained as single spots by thin-layer chromatography, produced a rise in [Ca2+]i in VSM cells from 45 +/- 7 to 99 +/- 22 and from 48 +/- 9 to 92 +/- 2 nmol/L (each n = 5; P < .05), respectively, after 3 min. V-diascorbate also increased [Ca2+]i slowly and dose-dependently, eg, from 56 +/- 14 to 102 +/- 15 nmol/L at a concentration of 10(-6) mol/L (n = 5; P < .05) after 3 min. A similar slow rise in [Ca2+]i from 53 +/- 10 to 185 +/- 3 nM (n = 5; P < .05) after 3 min was found with ouabain (10(-6) mol/L). As standard vasoconstrictor, All (10(-8) mol/L) rapidly increased [Ca2+]i from 23 +/- 4 to 846 +/- 50 nmol/L (n = 7; P < .01) within 30 sec. This effect was enhanced to 1,389 +/- 161 nM (n = 7; P < .01) when VSM cells were preincubated with V-diascorbate (10(-6) mol/L) for 10 min. AVP (10(-7) mol/L) also rapidly increased [Ca2+]i to 418 +/-11 nmol/L within 30 sec (n = 7; P < .01). This effect was enhanced in the presence of OLF-2 (approximately equals 10(-4) mol/L) or ouabain (10(-6) mol/L) to 523 +/- 14 and 560 +/- 19 nmol/L, respectively (each n = 7); P < .01). The calcium channel blocker verapamil, the intracellular calcium release blocker TMB-8, and the unselective cation channel blocker Ni2+ partly blunted the A II- or AVP-induced rise in [Ca2+]i and prevented the OLF-2- and V-diascorbate-induced increase in [Ca2+]i. Thus, OLF-1, OLF-2 and V-diascorbate, the active component of OLF-2, reveal effects similar to those of ouabain on [Ca2+]i in VSM cells, ie, they produce a slow rise in [Ca2+]i subsequent to inhibition of the sodium pump. The physiologic and pathologic roles of these and additional OLF in body fluid and blood pressure regulation and in hypertension have yet to be evaluated. PMID- 10821339 TI - Role of insulin in the regulation of membrane fluidity of erythrocytes in essential hypertension: an electron paramagnetic resonance investigation. AB - In the present study, to determine a possible role of insulin in the regulation of membrane functions, we have examined the effects of insulin on the membrane fluidity of erythrocytes in patients with essential hypertension and normotensive subjects. Membrane fluidity of erythrocytes obtained from hypertensive and normotensive subjects were evaluated by means of an electron paramagnetic resonance (EPR) and a spin-labeling method. In an in vitro study, insulin increased the order parameter (S for 5-nitroxide stearate) and the peak height ratio (ho/h-1 for 16 nitroxide stearate) in the EPR spectra of erythrocyte membranes, which indicated that insulin decreased the membrane fluidity of erythrocytes. The effects of insulin on the membrane fluidity were potentiated in the presence of extracellular Ca2+, and in contrast, were antagonized by the Ca2+ channel blocker diltiazem. Furthermore, the effects of insulin alone and in combination with Ca2+ on the membrane fluidity were reduced in the erythrocytes from hypertensive subjects compared with the erythrocytes from normotensive controls. The high concentrations of glucose alone produced no significant effects on the membrane fluidity of erythrocytes. These results demonstrated that insulin might actively participate in the regulation of membrane fluidity of erythrocytes, which might be mediated by the intracellular Ca2+ kinetics. PMID- 10821340 TI - Protein kinase C mediates insulin-inhibited Ca2+ transport and contraction of vascular smooth muscle. AB - Insulin acutely inhibits contraction of primary cultured vascular smooth muscle (VSM) cells from canine femoral artery by inhibiting contractile agonist-induced Ca2+ influx. Insulin also inhibits contraction at step(s) distal to intracellular Ca2+ concentration (Ca2+i) by stimulating cyclic guanosine monophosphate (GMP) production. We wished to see whether these effects of insulin are mediated by protein kinase C (PKC). Ca2+ influx was assessed by measuring the rate of fluorescence quenching of intracellular fura 2 by extracellular Mn2+. We found that 10 micromol/L serotonin (5-HT) stimulated Mn2+ influx 3-fold, and 1 nmol/L insulin inhibited the 5-HT-stimulated component of Mn2+ influx by 63% (P < .05), but insulin had no effect in the presence of 1 micromol/L staurosporine, an inhibitor of PKC. In the absence of insulin, preincubating cells with 0.1 micromol/L phorbol 12-myristate 13-acetate (PMA) for 5 min inhibited the 5-HT stimulated component of Mn2+ influx by 69% (P < .05). Insulin inhibited cell contraction induced by raising Ca2+i to supraphysiologic levels with ionomycin by 75% (P < .05). We also noted that 10(-6) mol/L calphostin C, another PKC inhibitor, or 16-h preincubation with PMA completely blocked this effect of insulin. Finally, 10-min exposure to insulin or PMA increased cyclic GMP production in ionomycin-treated cells by 50% and 64%, respectively (both P < .05). We conclude that insulin inhibits VSM cell contraction by inhibiting 5-HT stimulated Ca2+ influx and also at step(s) distal to Ca2+i by a PKC-dependent mechanism. PMID- 10821341 TI - Short-term angiotensin converting enzyme inhibition reduces basal tone and dilator reactivity in skeletal muscle arterioles. AB - Alterations in resting tone, maximum diameter, and dilator reactivity to acetylcholine (ACH) and sodium nitroprusside (SNP) were assessed in cremaster muscle microvessels of Sprague-Dawley rats receiving angiotensin converting enzyme (ACE) inhibition with captopril for 4 days and in untreated time-control rats. The transilluminated in situ cremaster muscle was superfused with physiologic salt solution (PSS) and viewed via television microscopy; arteriolar diameter was measured using a videomicrometer. Before agonist challenge, resting arteriolar diameter was significantly increased in captopril-treated rats. Although maximum arteriolar diameter (determined during superfusion of the cremaster muscle with Ca2+-free PSS containing 10(-4) mol/L adenosine) was not altered with ACE inhibition, the maximum possible arteriolar dilation was reduced in captopril-treated rats. Captopril administration reduced both ACH- and SNP induced dilation of cremasteric arterioles compared with responses in control rats, although this was partially a function of the reduced capacity for dilation, primarily to SNP. These observations indicate that short-term ACE inhibition reduces both resting tone and agonist-induced dilator responses of skeletal muscle arterioles. PMID- 10821343 TI - Cyclooxygenases 1 and 2 and thromboxane synthase in kidneys of Lyon hypertensive rats. AB - Compared with their two normotensive (LN and LL) controls, genetically hypertensive rats of the Lyon strain (LH) exhibit increased renal vascular resistance and a blunted pressure natriuresis function as well as an increased urinary excretion of vasoconstrictor prostanoids. The aim of this study was to assess in the kidneys of these animals the synthesis of vasoconstrictor or sodium retaining prostanoids. The relative abundance of the mRNAs of cyclooxygenases (COX) 1 and 2 and of thromboxane A2 synthase (TXS), was measured by reverse transcription polymerase chain reaction (RT-PCR) in renal cortex and medulla dissected in groups of male LH, LN, and LL rats either in baseline conditions or after 1 week of salt loading (1.5% NaCl in the drinking water). In basal conditions, at 3 and 11 weeks of age COX1 was expressed in the kidneys of all rats more markedly in medulla than in cortex. COX2 was poorly expressed in the whole kidney. TXS expression was usually too low to be quantified. No difference could be observed among LH, LN, and LL rats. After salt loading, the expression of COX1 was enhanced in the medulla and that of COX2 reduced in the cortex. LH rats differed from controls by a significantly more marked increase in medullary COX1 expression. The present work excludes any primary generalized increase in the renal expression of the genes that control the synthesis of vasoconstrictor prostanoids in LH rats, but suggests that medullary COX1 is upregulated by salt in these animals. PMID- 10821342 TI - Vitamin D analogs modulate the action of gonadal steroids in human vascular cells in vitro. AB - We have previously reported that estradiol (E2) and dihydrotestosterone (DHT) regulate cell growth in human umbilical arterial smooth muscle cells (SMC) and in an endothelial cell line (E304). In SMC both gonadal steroids stimulated DNA synthesis at low concentrations and suppressed 3[H] thymidine incorporation at high concentrations, whereas in E304 cells E2 and DHT dose dependently enhanced DNA synthesis. In both cell types gonadal steroids also induced the specific activity of creatine kinase BB (CK). Previous evidence suggets that the in vitro and in vivo CK responses to gonadal steroids in bone cells are upregulated by pretreatment with vitamin D analogs due to increased level of cellular estrogen receptors (ER). Here we analyzed the interaction of the vitamin D analogs hexafluorovitamin D (FL), JK-1624 F2-2 (JKF), and CB 1093 (CB) with gonadal steroids in regulating DNA synthesis and CK activity in human vascular cells in vitro. In E304 cells, daily treatment with FL, JKF, or CB (1 nmol/L for 3 days) increased DNA synthesis by 110 +/- 11%, 65 +/- 16%, and 88 +/- 23% respectively. In contrast, the same analogs inhibited 3[H] thymidine incorporation by 52 +/- 21%, 46 +/- 19%, and 50 +/- 10%, respectively, in SMC. In both cell types all three analogs increased CK by 25% to 75% and amplified the CK response to E2 and to DHT by twofold to threefold. In E304 cells the vitamin D analogs also increased DNA response to gonadal steroids from 50% to 60% to 200% to 280%. In SMC these analogs did not modify the DNA synthetic response to a low E2 concentration, but prevented the suppression of DNA synthesis exerted by high concentrations of E2 and DHT. Vitamin D inhibitors known to block cellular calcium mobilization, had no effect on the proliferative activity induced by vitamin D analogs. However, the inhibitor of the nuclear effects of vitamin D, ZK 159222, blocked the stimulatory effects of CB on DNA synthesis in E304 cells. Finally, both 1,25(OH)2 D3, and JKF decreased the expression of ERbeta proteins in SMC and increased the ERalpha isoform in E304 cells by 40% to 75%. The results indicate that vascular cells are targets for both vitamin D and gonadal steroid action and suggest a possible interaction between these hormones in the regulation of cell proliferation via modulation of vascular ER or interaction with proteins associated with ER. PMID- 10821344 TI - Corticosteroid dynamics in the nonpregnant, pregnant, and postpartum spontaneously hypertensive rat. AB - Factors responsible for hypertension in the spontaneously hypertensive rat (SHR) remain under investigation. As in human pregnancy complicated by essential chronic hypertension, the hypertension of the pregnant SHR subsides and returns postpartum. Because corticosteroid excess can cause hypertension, we examined several aspects of adrenocortical activity as potentially affecting the reported blood pressure profiles of nonpregnant, term pregnant, and postpartum SHR, using normotensive Wistar-Kyoto (WKY) rats as controls. We found that corticosterone levels were comparable in nonpregnant SHR and WKY rats, and unaffected by pregnancy. No differences were detected postpartum. Although pregnancy was accompanied by significant increases in plasma aldosterone levels, no interbreed differences were observed, which remained the case postpartum. Single adrenal cell secretion of aldosterone and corticosterone, as detected by reverse hemolytic plaque assay, yielded similar results in the pregnant and postpartum rat. Hormone responses to dietary manipulations in the nonpregnant and pregnant SHR and WKY suggest an important role for ACTH, and a lesser one for AII in the regulation of corticosteroids. In situ hybridization histochemistry, using a probe that detects both P450c11beta and P450c11AS mRNA, revealed comparable message density and zonal distribution in adrenals from pregnant and nonpregnant SHR and WKY rats. Breed- and pregnancy-dependent differences in adrenal expression of P450scc, P450c11beta, and P450c11AS were noted. In summary, our findings suggest that although some discrepancies exist in the aspects of adrenocortical activity examined, they are unlikely to be etiologic in the blood pressure profile observed in nonpregnant, pregnant, and postpartum SHR. PMID- 10821345 TI - Angiotensin II antagonists for hypertension: are there differences in efficacy? AB - We compared the antihypertensive efficacy of available drugs in the new angiotensin-II-antagonist (AIIA) class. The antihypertensive efficacy of losartan, valsartan, irbesartan, and candesartan was evaluated from randomized controlled trials (RCT) by performing a metaanalysis of 43 published RCT. These trials involved AIIA compared with placebo, other antihypertensive classes, and direct comparisons between AIIA. A weighted-average for diastolic and systolic blood pressure reduction with AIIA monotherapy, dose titration, and with addition of low-dose hydrochlorothiazide (HCTZ) were calculated. Weighted-average responder rates were also determined. The metaanalysis assessed a total of 11,281 patients. The absolute weighted-average reductions in diastolic (8.2 to 8.9 mm Hg) and systolic (10.4 to 11.8 mm Hg) blood pressure reductions (not placebo corrected) for AIIA monotherapy were comparable for all AIIA. Responder rates for AIIA monotherapy were 48% to 55%. Dose titration resulted in slightly greater blood pressure reduction and an increase in responder rates to 53% to 63%. AIIA/hydrochlorothiazide combinations produced substantially greater reduction in systolic (16.1 to 20.6 mm Hg) and diastolic (9.9 to 13.6 mm Hg) blood pressure reductions than AIIA monotherapy and responder rates for AIIA/HCTZ combinations were 56% to 70%. This comprehensive analysis shows comparable antihypertensive efficacy within the AIIA class, a near-flat AIIA-dose response when titrating from starting to maximum recommended dose, and substantial potentiation of the antihypertensive effect with addition of HCTZ. PMID- 10821346 TI - Intracellular Mg++ concentrations in smooth and striated muscle cells in spontaneously hypertensive rats. AB - Decreased intracellular Mg++ concentrations seem to be involved in the pathogenesis of primary hypertension. Of special interest is the smooth muscle cell with its electrolyte metabolism in primary hypertension, but also heart muscle cells and their Mg++ concentrations are of growing interest. Therefore, in aortic smooth muscle cells and striated heart muscle cells (left ventricle) from 20 spontaneously hypertensive rats (SHR) of the Munster strain and 20 normotensive Wistar-Kyoto rats (WKY), the intracellular Mg++ content was measured. The electron probe x-ray microanalysis technique was used to determine intracellular Mg++ concentrations under nearly in vivo conditions in aortic cryosections 3 microm thick and striated heart muscle cells 4 microm thick (Camscan CS 24 apparatus). Vascular smooth muscle Mg++ content was 36.4 +/- 3.1 mmol/kg dry weight in SHR versus 48.6 +/- 3.7 mmol/kg dry weight in WKY (P < .001). In striated heart muscle cell Mg++ concentrations, there was no significant difference in SHR and WKY (79.9 +/- 5.6 versus 80.3 +/- 5.9 mmol/ kg dry weight). In conclusion, the present study revealed that genetic hypertension in the spontaneously hypertensive rat is accompanied by significantly decreased intracellular Mg++ concentrations in vascular smooth muscle cells. In striated heart muscle cells, Mg++ content was not significantly different in SHR and WKY. Mg++ handling is different in vascular smooth muscle and striated heart muscle cells in WKY and SHR (P < .01). PMID- 10821347 TI - Loperamide therapy in a patient with diarrhea and resistant hypertension. AB - A 65-year-old man was admitted to our hospital for resistant essential hypertension. He had a past history of bowel resections due to Crohn's disease. We assumed that in this patient insufficient drug absorption might be a cause of resistant hypertension and that even increased antihypertensive regimens would fail to decrease blood pressure. We administered loperamide, which may increase drug absorption from the residual intestine. Blood pressure was successfully decreased, with a concomitant increase in blood concentration of antihypertensive drugs. In patients with impaired drug absorption, we should make an effort to restore drug obstruction before considering increased drug regimens. PMID- 10821348 TI - The deletion polymorphism of the angiotensin-converting enzyme is associated with nephroangiosclerosis. AB - The D allele of the angiotensin-converting enzyme (ACE) gene has been linked with diabetic nephropathy and IgA glomerulonephritis and with faster renal disease progression. The association of this allele with nephroangiosclerosis has been scarcely investigated. We have tested this association in 45 hypertensive patients (all whites) with well defined nephroangiosclerosis (diagnosis established on the basis of renal biopsy in all cases) and moderate to severe renal failure. As studies of genetic association of small size often produce conflicting results, besides a control group of 343 Italian patients with essential hypertension and normal renal function, we elected to use also a very large control group of race-matched subjects taken from a meta-analysis of 27,565 whites. The proportion of patients with the D allele (64%) was higher in patients with nephroangiosclerosis than that in Italian hypertensives (54%) and in whites (54%). DD and DI genotypes were more prevalent in patients than in control groups. The dominant model (DD and DI v II: nephroangiosclerosis v Italian controls: chi2 = 6.19, P = .012; nephroangiosclerosis v whites chi2 = 6.86, P = .009) fitted the data better than the codominant and the recessive model (P < or = .022). The D allele is associated with nephroangiosclerosis with a dominant effect in the sample of patients studied. Although intervention studies are needed to see whether these findings imply a causal association, our data suggest that this allele may at least act as disease marker in nephroangiosclerosis. PMID- 10821349 TI - Distribution of different HLA antigens in Greek hypertensives according to the angiotensin-converting enzyme genotype. AB - The angiotensin-converting enzyme (ACE) insertion/deletion polymorphism is an independent risk factor for cardiovascular disease. It has also been suggested that some HLA genes may contribute to the genetic susceptibility to essential hypertension. So far, an association between ACE polymorphism and HLA antigens in arterial hypertension has not been reported. We have studied 94 subjects with newly diagnosed essential hypertension, 49 men and 45 women (mean age, 52.3 +/- 11.3 years), as well as 104 randomly selected, age- and gender-matched normotensive individuals (54 men and 50 women, mean age 48.7 +/- 10.8 years). Both cohorts originated from the Greek population and lived in the greater Athens area. The ACE genotype was analyzed by polymerase chain reaction. HLA class I and II antigens were studied by serologic and molecular techniques. The prevalence of the ACE genotypes did not differ significantly between hypertensives and normal individuals. The casual blood pressure levels and the average ambulatory blood pressure levels were similar among the three ACE genotypes. Hypertensives with the ACE-DD genotype were characterized by an increased prevalence of the HLA-A2 antigen (50% v 31.4%, P < .005) and DR6 (16.7% v 11.4%, P < .01) in comparison to the normotensive subjects with the ACE-DD genotype. HLA-A24 was found more frequently among the hypertensives with the ACE-ID genotype than in the normal controls with the same genotype (35.5% v 26.4%, P < .05). ACE-DD genotype is associated with a high prevalence of specific HLA antigens. The coexistence of the ACE-DD genotype with certain HLA phenotypes could reveal a distinct hypertensive population with increased risk for cardiovascular events. PMID- 10821350 TI - Angiotensin receptors: history and mysteries. AB - Angiotensin receptors became relatively easy to study when radioactive derivatives of the peptide were synthesized for radioimmunoassays. Binding assays in vitro led to the discovery of receptors in many tissues different from those involved in the classic actions of angiotensin. The physiologic significance of receptors in sites such as the gonads, other endocrine organs, peripheral blood cells, and many regions of the brain is still uncertain. Kinetics of the binding reaction are susceptible to intracellular guanine nucleotides, and extracellular cations, fatty acids, steroids, and eicosanoids. Synthesis of receptors is under equally complex control. Receptor binding assays simplified screening for angiotensin antagonists. Nonpeptide antagonists proved so specific they revealed the existence of receptor subtypes. The two principal subtypes are found in different tissues and trigger different postreceptor cascades. Studies of receptors, the genes that code for them, and the drugs that block them have led to a growing awareness of angiotensin's effects on the structure of the heart, vessels, and kidneys, some of which are pathologic. The existence of receptor subtypes, the different signal transduction cascades they stimulate, the widespread location of receptors, and the range of effects they mediate suggest that the angiotensins are of broad relevance in biology and pathology. This multidimensional matrix also indicates that receptor antagonists may have effects not yet described. PMID- 10821351 TI - Cardiovascular remodeling, apoptosis, and drugs. AB - Apoptosis, a form of programmed cell death, mediates the controlled deletion of so-called "unwanted" cells. This review deals with the key features of this cell death program, showing that apoptosis is regulated by factors extrinsic and intrinsic to the dying cell. The elucidation of the possible interactions between these factors may be of major interest in preventing the progression to cardiovascular remodeling in patients with hypertensive disease. New pathways of research are emerging for drugs, such as beta-blockers, ACE inhibitors, the calcium-antagonists, and the receptor antagonist of angiotensin II, all of which have beneficial effects on cardiovascular remodeling. This may be due to the direct effect of these drugs on the cell proliferation/apoptosis balance. PMID- 10821352 TI - Effect of ethanol on blood pressure--role of magnesium. PMID- 10821353 TI - Comparisons in a competitive world: when is one drug superior to another? PMID- 10821354 TI - The future for vCJD. PMID- 10821355 TI - All that glitters is not gold. PMID- 10821356 TI - Microgravity-induced bone loss--will it limit human space exploration? PMID- 10821357 TI - Water channels: who needs them anyway? PMID- 10821358 TI - Steroid therapy for visual loss in patients with giant-cell arteritis. PMID- 10821359 TI - Academia and industry: increasingly uneasy bedfellows. PMID- 10821360 TI - Long-term ACE-inhibitor therapy in patients with heart failure or left ventricular dysfunction: a systematic overview of data from individual patients. ACE-Inhibitor Myocardial Infarction Collaborative Group. AB - BACKGROUND: We undertook a prospective systematic overview based on data from individual patients from five long-term randomised trials that assessed inhibitors of angiotensin-converting enzyme (ACE) in patients with left ventricular dysfunction or heart failure. METHODS: Three of the trials enrolled patients within a week after acute myocardial infarction. Data were combined by use of the Peto-Yusuf method. FINDINGS: Overall 12,763 patients were randomly assigned treatment or placebo and followed up for an average of 35 months. In the three post-infarction trials (n=5,966), mortality was lower with ACE inhibitors than with placebo (702/2995 [23.4%] vs 866/2971 [29.1%]; odds ratio 0.74 [95% CI 0.66-0-83]), as were the rates of readmission for heart failure (355 [11.9%] vs 460 [15.5%]; 0.73 [0.63-0.85]), reinfarction (324 [10.8%] vs 391 [13.2%]; 0.80 [0.69-0.94]), or the composite of these events (1049 [35.0%] vs 1244 [41.9%]; 0.75 [0.67-0.83]; all p palmitate > oleate), but had little effect on ketogenesis from medium chain fatty acids (octanoate and laureate). With palmitate the decrease in oxygen uptake was restricted to the substrate stimulated portion; with stearate, the decrease exceeded the substrate stimulated portion; with oleate, oxygen uptake was transiently inhibited. Withdrawal of Ca2+ attenuated the inhibitory effects. 14CO2 production from [1-14C]oleate was inhibited. Net uptake of the fatty acids was not affected by norepinephrine. In livers from fed rats, oxygen uptake and ketogenesis from stearate were only transiently inhibited. The conclusions are: (a) in the fasted state norepinephrine reduces ketogenesis and respiration by means of a Ca2+-dependent mechanism; (b) the degree of inhibition varies with the chain length and the degree of saturation of the fatty acids; (c) norepinephrine favours esterification of the activated long-chain fatty acids in detriment to oxidation; (d) in the fed state the stimulatory action of norepinephrine on glycogen catabolism induces conditions which are able to reverse inhibition of ketogenesis and oxygen uptake. PMID- 10821420 TI - Involvement of calcium in the stimulation of phosphatidylcholine secretion in primary cultures of rat type II pneumocytes by Escherichia coli lipopolysaccharide. AB - The purpose of this study was to evaluate the mechanism by which Escherichia coli lipopolysaccharide stimulates the secretion of phosphatidylcholine in primary cultures of rat type II pneumocytes. The stimulatory effect of lipopolysaccharide on phosphatidylcholine secretion was additive to those of terbutaline and TPA (protein kinase A and C activators respectively) and this effect was not suppressed by inhibitors of both protein kinases. On the other hand, lipopolysaccharide did not modify the increase on phosphatidylcholine secretion induced by the endoplasmic reticulum Ca2+-ATPase inhibitor thapsigargin, and enhanced slightly the calcium-ionophore A23187 stimulated phosphatidylcholine secretion. In addition, the stimulatory effect of lipopolysaccharide was suppressed by BAPTA, an intracellular Ca2+ chelator, and KN-62, a specific inhibitor of Ca2+-calmodulin-dependent protein kinase. These results, together with the lipopolysaccharide-mediated increase in the cytosolic [Ca2+], suggest that stimulation of phosphatidylcholine secretion by lipopolysaccharide in type II pneumocytes occurs by a calcium-dependent transduction mechanism via Ca2+ calmodulin-dependent protein kinase activation. PMID- 10821419 TI - Molecular characterization of anion exchangers in the cochlea. AB - Anion exchange proteins (AE) in the inner ear have been the focus of attention for some time. They have been suggested to play a role as anion exchangers for the regulation of endolymphatic pH or as anion exchangers and anchor proteins for the maintenance of the shape and turgor of outer hair cells, and they also have been discussed as a candidate protein for motile hair cell responses that follow high-frequency stimulation. The existence of anion exchangers in hair cells and the specific isoforms which are expressed in hair cells and the organ of Corti is controversial. Using a polyclonal antibody to AE1 (AB 1992, Chemicon), we immunoprecipitated a 100 kDa AE polypeptide in isolated outer hair cells which, due to its glycosylation, is comprised of AE2 than AE1 isoforms. We confirmed AE2 expression in outer hair cells with the help of subtype-specific monoclonal and polyclonal antibodies to AE, AE subtype-specific primers and AE subtype-specific cDNA and found glycosylated truncated as well as full-length AE2 isoforms. No AE1 or AE3 subtypes were noted in outer hair cells. In contrast, AE2 and AE3 but not AE1 subtypes were seen in supporting cells of the organ of Corti. Their expression preceded the development of cochlear function, coincident with the establishment of the endocochlear potential and the differentiation of supporting cells. While most developmental processes in the inner ear usually begin in the basal cochlear turn, the AE2 expression in outer hair cells (but not that of AE2 and AE3 in supporting cells) progressed from the apical to the basal cochlear turn, reminiscent of the maturation of frequency-dependency. Irrespective of their presumed individual role as either anion exchanger, anchor protein or motility protein, the differential expression and developmental profile of these proteins suggest a most important role of anion exchange proteins in the development of normal hearing. These findings may also provide novel insights into AE function in general. PMID- 10821421 TI - Differential effect of fasting on IGF-BPs in serum of young and adult rats and its implication to impaired skin GAG content. AB - During fasting or aging of animals there is a decreased content of skin glycosaminoglycans (GAGs). It has been found that the skin of adult rats contains about 60% of GAGs found in the skin of young animals. Fasting of both groups of animals (young and adult) resulted in decrease of GAG content. However, GAG content in the skin of fasted young rats decreased by 30% and in fasted adult rats by 15% only, compared to fed animals, respectively. The mechanism for the phenomena is not known. We considered insulin-like growth factor-I (IGF-I) as a potential candidate involved in regulation of GAG biosynthesis in both experimental models of animals. Adult rat sera were found to contain about 75% of IGF-I recovered from young rat sera. Fasting of both groups of animals resulted in dramatic decrease in serum IGF-I levels to about 50% of initial values. Since IGF-I activity and IGF-I serum half-life depends on the level of specific IGF binding proteins (IGFBPs) we determined (i) relationship between main groups of IGFBPs, namely high molecular weight binding proteins (HMWBPs) and low molecular weight binding proteins (LMWBPs) and (ii) the amounts of IGF-I bound to respective proteins in the sera of all experimental animals. Control young rat serum was found to contain about 90% of HMWBPs and about 10% of LMWBPs as determined by ligand binding assay. In contrast, control adult rat serum contained about 60% of HMWBPs and about 40% of LMWBPs. Fasting of both groups of animals resulted in significant increase in serum levels of LMWBPs. Control young rat serum was found to contain about 8% IGF-I bound to LMWBPs while serum of control adult rats contained 18% IGF-I bound to these proteins. In sera of fasted young animals however, about 75% of the bound IGF-I was recovered from LMWBPs (about 60% of total serum IGF-I) while in sera of fasted adult animals only about 56% of the bound IGF-I was recovered from LMWBPs (about 50% of total serum IGF I). Evidence was provided that during fasting of both groups of animals there is a significant decrease in serum BP-3 and dramatic increase in serum BP-1 concentrations, compared to respective controls. However, the concentration of BP 1 in serum of fasted young rats was increased by about 60 fold while in serum of fasted adult rats only by about 10 fold, compared to respective control animals. Negative correlation between skin GAG content and LMWBPs derived IGF-I during fasting of young (r = -0.943, p < 0.001) and adult ( r = -0.571, p < 0.01) rats was found. The data presented suggest that the effects of aging and fasting on decreased skin GAG content may be due to induction of LMWBPs that are known to (i) inhibit IGF-I dependent function and (ii) increase clearance of IGF-I from circulation. However, the effects of fasting are distinct in respect to young and adult rats suggesting that mechanisms involved in regulation of IGF-I bioactivity during aging are more complex that during fasting. PMID- 10821422 TI - Evidence for the induction of apoptosis by endosulfan in a human T-cell leukemic line. AB - Several organochlorinated pesticides including DDT, PCBs and dieldrin have been reported to cause immune suppression and increase susceptibility to infection in animals. Often this manifestation is accompanied by atrophy of major lymphoid organs. It has been suggested that increased apoptotic cell death leading to altered T-B cell ratios, and loss of regulatory cells in critical numbers leads to perturbations in immune function. The major objective of our study was to define the mechanism by which endosulfan, an organochlorinated pesticide, induces human T-cell death using Jurkat, a human T-cell leukemic cell line, as an in vitro model. We exposed Jurkat cells to varying concentrations of endosulfan for 0-48 h and analyzed biochemical and molecular features characteristic of T-cell apoptosis. Endosulfan lowered cell viability and inhibited cell growth in a dose- and time-dependent manner. DAPI staining was used to enumerate apoptotic cells and we observed that endosulfan at 10-200 microM induced a significant percentage of cells to undergo apoptotic cell death. At 48 h, more than 90% cells were apoptotic with 50 microM of endosulfan. We confirmed these observations using both DNA fragmentation and annexin-V binding assays. It is now widely being accepted that mitochondria undergo major changes early during the apoptotic process. We examined mitochondrial transmembrane potential (deltapsim) in endosulfan treated cells to understand the role of the mitochondria in T-cell apoptosis. Within 30 min of chemical exposure, a significant percentage of cells exhibited a decreased incorporation of DiOC6(3), a cationic lipophilic dye into mitochondria indicating the disruption of deltapsim. This drop in deltapsim was both dose- and time-dependent and correlated well with other parameters of apoptosis. We also examined whether this occurred by the down regulation of bcl-2 protein expression that is likely to increase the susceptibility of Jurkat cells to endosulfan toxicity. Paradoxically, the intracellular expression of bcl-2 protein was elevated in a dose dependent manner suggesting endosulfan-induced apoptosis occurred by a non-bcl-2 pathway. Based on these data, as well as those reported elsewhere, we propose the following sequence of events to account for T cell apoptosis induced by endosulfan: uncoupling of oxidative phosphorylation --> excess ROS production --> GSH depletion --> oxidative stress --> disruption of deltapsim --> release of cytochrome C and other apoptosis related proteins to cytosol --> apoptosis. This study reports for the first time that endosulfan can induce apoptosis in a human T-cell leukemic cell line which may have direct relevance to loss of T cells and thymocytes in vivo. Furthermore, our data strongly support a role of mitochondrial dysfunction and oxidative stress in endosulfan toxicity. PMID- 10821423 TI - Down regulation of myocardial beta1-adrenoceptor signal transduction system in pacing-induced failure in dogs with aortic stenosis-induced left ventricular hypertrophy. AB - We recently demonstrated that rapid ventricular pacing caused cardiac failure (Failure) in dogs with aortic stenosis-induced left ventricular hypertrophy (Hypertrophy) and isoproterenol caused no significant increases in function, O2 consumption and intracellular cyclic AMP level in the failing hypertrophied hearts. We tested the hypothesis that alterations in the beta1-adrenoceptor signal transduction pathway would correlate with the reduced functional and metabolic responses to beta-adrenergic stimulation during the transition from the compensated hypertrophy to failure. Pressure overload-induced left ventricular hypertrophy was created using aortic valve plication in 10 dogs over a 6-month period. Five months after aortic valve plication, congestive heart failure was induced in 5 dogs by rapid ventricular pacing at 240 bpm for 4 weeks. The density of myocardial beta1-adrenoceptors (fmoles/mg membrane protein; fmoles/g wet tissue) was significantly reduced in the Failure dogs (176+/-19; 755+/-136) when compared to those of the Control (344+/-51; 1,551+/-203) and the Hypertrophy (298+/-33; 1,721+/-162) dogs. The receptor affinities were not significantly different among all groups. There was a small but significant decrease in the percentage of beta1-adrenoceptors of the failing hypertrophied hearts (62+/-3%) when compared to that of the hypertrophied hearts (77+/-5%). The basal myocardial adenylyl cyclase activity (pmoles/mg protein/min) was significantly lower in the Failure dogs (45+/-4) than in the Control (116+/-14) and Hypertrophy (86+/-6) dogs. The forskolin (0.1 mM)-stimulated adenylyl cyclase activity was also significantly lower in the Failure dogs (158+/-17) than in the Control dogs (296+/-35) and slightly lower than in the Hypertrophy dogs (215+/-10). There were no significant differences in low Km cyclic AMP-phosphodiesterase activities among all groups. We conclude that down regulation of beta1-adrenoceptors and reduced adenylyl cyclase activities contribute to the decreases in myocardial functions and beta-adrenergic responses in the failing hypertrophied hearts induced by rapid ventricular pacing. PMID- 10821424 TI - Characterization of sphingomyelinase activity released by thrombin-stimulated platelets. AB - In this study we report that human platelets display neutral (nSMase) and acid sphingomyelinase (aSMase) as well as acid ceramidase (aCerase) activity. Cell activation by thrombin resulted in a marked decrease of intracellular aSMase activity, accompanied by the release of enzyme into the medium. In contrast, thrombin treatment did not affect aCerase activity. Two major protein bands of 73 and 70 kDa were recognized by aSMase antibodies in resting platelet lysates and in the medium of stimulated cells. Phorbol esters together with the calcium ionophore A23187 fully reproduced thrombin action on aSMase release. The secreted enzymatic activity was insensitive to digestion with endoglycosidase H but it was stimulated by Zn2+, although to a limited extent compared to aSMase constitutively released by murine endothelial cells. Taken together, these data suggest that secreted aSMase does not originate from the lysosomal compartment but rather from other platelet vesicles. PMID- 10821425 TI - Sterol carrier protein-2 suppresses microsomal acyl-CoA hydrolysis. AB - Although sterol carrier protein 2 (SCP-2) has long been regarded primarily as a sterol transfer protein, its actual physiological function is not known. The recent discovery that SCP-2 binds long chain fatty acyl-CoAs (LCFA-CoAs) with high affinity suggests additional roles for SCP-2 in cellular utilization of LCFA CoAs for synthesis of glycerides and cholesterol esters. Concomitant to these anabolic pathways, LCFA-CoAs are also degraded by cellular hydrolases. The purpose of the work presented herein was to determine if SCP-2 altered the aqueous pool of LCFA-CoA by (i) extracting LCFA-CoA from microsomal membranes, and (ii) protecting LCFA-CoA from microsomal hydrolase activity. The data demonstrated for the first time that SCP-2 increases the aqueous pool of oleoyl CoA by increasing the aqueous/membrane distribution oleoyl-CoA by 2.4-fold. In addition, SCP-2 inhibited the hydrolysis of oleoyl-CoA by microsomal acyl-CoA hydrolase 1.6-2.4 fold, depending on the concentration of oleoyl-CoA. By simultaneously extracting LCFA-CoA from membranes and inhibiting LCFA-CoA degradation SCP-2 may potentiate LCFA-CoA transacylation and modulate the role of LCFA-CoAs as intracellular signaling molecules. PMID- 10821426 TI - The analysis of the poly(ADPR) polymerase mode of action in rat testis nuclear fractions defines a specific poly(ADP-ribosyl)ation system associated with the nuclear matrix. AB - The poly(ADP-ribosyl)ation system, associated with different nuclear fractions of rat testis, has been analyzed for both pADPR and pADPR acceptor proteins. The DNase I sensitive and resistant chromatin contain 35% and 40%, respectively, of the total pADPR synthesized in intact nuclei incubated with [32P]NAD. Moreover, the residual 25% were estimated to be associated with the nuclear matrix. Three different classes of pADPR are present in the nuclei. The longest and branched ADPribose polymers modify proteins present in the DNase I resistant (2 M NaCl extractable) chromatin and in the nuclear matrix, whereas polymers of> 20 residues interact with the components of the DNase I sensitive chromatin and oligomers of 6 ADPribose residues are bound specifically to the acid-soluble chromosomal proteins, present in isolated nuclear matrix. The main pADPR acceptor protein in all the nuclear fractions is represented by the PARP itself (auto modification reaction). The hetero-modification reaction occurs mostly on histone H1 and core histones, that have been found associated to DNase I sensitive and resistant chromatin, respectively. Moreover, an oligo(ADP-ribosyl)ation occurs on core histones tightly-bound to the matrix associated regions (MARs) of chromatin loops. PMID- 10821427 TI - Developmental regulation of the L-type calcium channel alpha1C subunit expression in heart. AB - We used Northern analyses, RNase protection assays and immunoblot analyses to examine the relationship among developmental age of the heart, abundance of mRNA and L-type calcium channel alpha1C subunit protein, and to establish the size of the native protein in heart. Northern analysis, RNase protection assays, and immunoblots were used to study RNA and protein from rat heart of various ages. In fetal and adult ventricles there was a predominant 8.3-kb transcript for the alpha1C subunit with no change in transcript size during development. RNase protection assays demonstrated a 2-fold increase in abundance of the DHP receptor message during postnatal development. Immunoblots identified a 240 kD protein, corresponding to the predicted molecular mass of the full length alpha1C subunit. No change in size of protein for the alpha1C subunit was observed at any developmental stage and there was no evidence for a truncated isoform. There was an approximate 2-fold increase in alpha1C subunit protein in ventricular homogenates during postnatal development. Thus, in the developing rat heart, alterations in calcium channel properties during development appear to result neither from alternative splicing that produces a smaller transcript for the alpha1C subunit nor from expression of a truncated protein, but at least in part from transcriptionally-regulated expression of the 240 kDa polypepde. PMID- 10821428 TI - Differential effect of schisandrin B and dimethyl diphenyl bicarboxylate (DDB) on hepatic mitochondrial glutathione redox status in carbon tetrachloride intoxicated mice. AB - The effects of schisandrin B (Sch B), a dibenzocyclooctadiene derivative isolated from the fruit of Schisandra chinensis, and dimethyl diphenyl bicarboxylate (DDB), a synthetic intermediate of schisandrin C (also a dibenzocyclooctadiene derivative), on hepatic mitochondrial glutathione redox status in control and carbon tetrachloride (CCl4)-intoxicated mice were examined. Treating mice with Sch B or DDB at a daily oral dose of 1 mmol/kg for 3 d did not produce any significant alterations in plasma alanine aminotransferase (ALT) and sorbital dehydrogenase (SDH) activities. CCl4 treatment caused drastic increases in both plasma ALT and SDH activities in mice. Pretreating mice with Sch B or DDB at the same dosage regimen significantly suppressed the CCl4-induced increase in plasma ALT activity, with the inhibitory effect of Sch B being much more potent. Sch B, but not DDB, pretreatment could also decrease the plasma SDH activity in CCl4 intoxicated mice. The lowering of plasma SDH activity, indicative of hepatoprotection against CCl4 toxicity, by Sch B pretreatment was associated with an enhancement in hepatic mitochondrial glutathione redox status as well as an increase in mitochondrial glutathione reductase (mtGRD) activity in both non-CCl4 and CCl4-treated mice. DDB pretreatment, though enhancing both hepatic mitochondrial glutathione redox status and mtGRD activity in control animals, did not produce any beneficial effect in CCl4-treated mice. The difference in hepatoprotective action against CCl4 toxicity between Sch B and DDB may therefore be related to their ability to maintain hepatic mitochondrial glutathione redox status under oxidative stress condition. PMID- 10821429 TI - Mechanisms of agonist-dependent and -independent desensitization of a recombinant P2Y2 nucleotide receptor. AB - UTP activates P2Y, receptors in both 1321N1 cell transfectants expressing the P2Y2 receptor and human HT-29 epithelial cells expressing endogenous P2Y, receptors with an EC50 of 0.2-1.0 microM. Pretreatment of these cells with UTP diminished the effectiveness of a second dose of UTP (the IC50 for UTP-induced receptor desensitization was 0.3-1.0 microM for both systems). Desensitization and down-regulation of the P2Y2 nucleotide receptor may limit the effectiveness of UTP as a therapeutic agent. The present studies investigated the phenomenon of P2Y2 receptor desensitization in human 1321N1 astrocytoma cells expressing recombinant wild type and C-terminal truncation mutants of the P2Y2 receptor. In these cells, potent P2Y2 receptor desensitization was observed after a 5 min exposure to UTP. Full receptor responsiveness returned 5-10 min after removal of UTP. Thapsigargin, an inhibitor of Ca2+-ATPase in the endoplasmic reticulum, induced an increase in the intracellular free calcium concentration, [Ca2+]i, after addition of desensitizing concentrations of UTP, indicating that P2Y2 receptor desensitization is not due to depletion of calcium from intracellular stores. Single cell measurements of increases in [Ca2+]i induced by UTP in 1321N1 cell transfectants expressing the P2Y2 receptor indicate that time- and UTP concentration-dependent desensitization occurred uniformly across a cell population. Other results suggest that P2Y2 receptor phosphorylation/dephosphorylation regulate receptor desensitization/resensitization. A 5 min preincubation of 1321N1 cell transfectants with the protein kinase C activator, phorbol 12-myristate 13 acetate (PMA), reduced the subsequent response to UTP by about 50%, whereas co incubation of PMA with UTP caused a greater inhibition in the response. The protein phosphatases-1 and -2A inhibitor, okadaic acid, partially blocked resensitization of the receptor. Furthermore, C-terminal truncation mutants of the P2Y2 receptor that eliminated several potential phosphorylation sites including two for PKC were resistant to UTP-, but not phorbol ester-induced desensitization. Down regulation of protein kinase C isoforms prevented phorbol ester-induced desensitization but had no effect on agonist-induced desensitization of wild type or truncation mutant receptors. These results suggest that phosphorylation of the C-terminus of the P2Y2 receptor by protein kinases other than protein kinase C mediates agonist-induced receptor desensitization. A better understanding of the molecular mechanisms of P2Y2 nucleotide receptor desensitization may help optimize a promising cystic fibrosis pharmacotherapy based on the activation of anion secretion in airway epithelial cells by P2Y, receptor agonists. PMID- 10821431 TI - Effects of HNS-32, a novel antiarrhythmic drug, on ventricular arrhythmias induced by coronary artery occlusion and reperfusion in anesthetized rats. AB - HNS-32 (N1,N1-dimethyl-N2-(2-pyridylmethyl)-5-isopropyl-3, 8-dimethylazulene-1 carboxamidine: CAS 186086-10-2) is a newly synthesized compound, and possesses antiarrhythmic properties with vasodilator action in dog hearts. The aim of this study was to investigate the dose-dependent effects of HNS-32 on ischemia- and/or reperfusion-induced ventricular arrhythmias in anesthetized rats in vivo and compared with those of mexiletine. Saline or drugs were administered intravenously 5 min prior to coronary artery occlusion. On the ischemia-induced ventricular arrhythmias, HNS-32 showed dose-dependent reduction of total number of premature ventricular complexes (PVC) from 2091+/-225 to 656+/-116 and 286+/ 69 beats/30 min (p < 0.05), the ventricular tachycardia (VT) duration from 183+/ 33 to 28+/-9 and 4+/-2 sec (p < 0.05), the incidence of VT from 100 to 90 (n.s.) and 40% (p < 0.05), and the incidence of ventricular fibrillation (VF) from 50 to 0 and 0% (p < 0.05) with 3 and 5 mg/kg, respectively. Mexiletine also reduced these parameters to 936+/-159 beats/30 min (p < 0.05), 39+/-22 sec (p < 0.05), 90% (n.s.) and 10% (n.s.), respectively. HNS-32 completely suppressed the late reperfusion-induced arrhythmias, however mexiletine did not affect them. On the early reperfusion-induced ventricular arrhythmias, HNS-32 showed dose-dependent reduction of VT duration from 126+/-34 to 37+/-12 and 3+/-2 sec (p < 0.05), incidence of VT from 100 to 90 (n.s.) and 40% (p < 0.05), incidence of VF from 100 to 10 and 0% (p < 0.05), and mortality rate from 90 to 0 and 0% (p < 0.05), with 3 and 5 mg/kg, respectively. Mexiletine also reduced these parameters to 16+/-9 sec (p < 0.05), 80 (n.s.), 50 (p < 0.05), and 10% (p < 0.05), respectively. HNS-32 significantly reduced the heart rate in a dose-dependent manner, from 399+/-14 to 350+/-8 and 299+/-10 beats/min (p < 0.05) with 3 and 5 mg/kg, respectively. The antiarrhythmic effects of HNS-32 were more potent than that of the similar dose of mexiletine against occlusion-induced and reperfusion induced arrhythmias in in vivo rats. PMID- 10821430 TI - Modulation of sulfated glycosaminoglycan and small proteoglycan synthesis by the extracellular matrix. AB - Cell culture in collagen lattice is known to be a more physiological model than monolayer for studying the regulation of extracellular matrix protein deposition. The synthesis of sulfated glycosaminoglycans (GAG) and dermatan sulfate (DS) proteoglycans by 3 cell strains were studied in confluent monolayers grown on plastic surface, in comparison to fully retracted collagen lattices. Cells were labelled with 35S-sulfate, followed by GAG and proteoglycan analysis by cellulose acetate and SDS-polyacrylamide gel electrophoresis, respectively. The 3 cell strains contracted the lattice in a similar way. In monolayer cultures, the major part of GAG was secreted into culture medium whereas in lattice cultures of dermal fibroblasts and osteosarcoma MG-63 cells but not fibrosarcoma HT-1080 cells, a higher proportion of GAGs, including dermatan sulfate, was retained within the lattices. Small DS proteoglycans, decorin and biglycan, were detected in fibroblasts and MG-63 cultures. They were preferentially trapped within the collagen gel. In retracted lattices, decorin had a higher Mr than in monolayer. Biglycan was detected in monolayer and lattice cultures of MG-63 cells but in lattice cultures only in the case of fibroblasts. In this last case, an up regulation of biglycan mRNA steady state level and down regulation of decorin mRNA was observed, in comparison to monolayers, indicating that collagen can modulate the phenotypical expression of small proteoglycan genes. PMID- 10821432 TI - Cell type-dependent transactivation or repression of mesoderm-restricted basic helix-loop-helix protein, POD-1/Capsulin. AB - A family of basic-helix-loop-helix (bHLH) nuclear factors play important roles in controlling cell growth and differentiation as critical regulatory components in transcription. Here we describe molecular characterization of mesoderm-specific bHLH protein, POD-1/Capsulin. Transactivation property of POD-1/Capsulin was analyzed by the Gal4 fusion system in six mammalian cell lines. The results indicated that an activation property was shown in HT1080 and HeLa cells, but a repression activity in HepG2 cells. Mapping analysis for the transactivation and repression activities revealed that the C-terminal domain of POD-1/Capsulin is essential for the transactivation and both the N-terminal and C-terminal domains are contributed to the repression activities. Furthermore, in order to identify possible interactants of the POD-1/Capsulin, we performed yeast two-hybrid screen in a human kidney cDNA library, and identified a class A bHLH protein, ITF-2 as potential heterodimeric partner of the bHLH protein. PMID- 10821434 TI - Effect of dephostatin on intracellular free calcium concentration and amylase secretion in isolated rat pancreatic acinar cells. AB - This study investigates the effects of dephostatin, a new tyrosine phosphatase inhibitor, on intracellular free calcium concentration ([Ca2+]i) and amylase secretion in collagenase dispersed rat pancreatic acinar cells. Dephostatin evoked a sustained elevation in [Ca2+]i by mobilizing calcium from intracellular calcium stores in either the absence of extracellular calcium or the presence of lanthanium chloride (LaCl3). Pretreatment of acinar cells with dephostatin prevented cholecystokinin-octapeptide (CCK-8)-induced signal of [Ca2+]i and inhibited the oscillatory pattern initiated by aluminium fluoride (AlF4), whereas co-incubation with CCK-8 enhances the plateau phase of calcium response to CCK-8 without modifying the transient calcium spike. The effects of dephostatin on calcium mobilization were reversed by the presence of the sulfhydryl reducing agent, dithiothreitol. Stimulation of acinar cells with thapsigargin in the absence of extracellular Ca2+ resulted in a transient rise in [Ca2+]i. Application of dephostatin in the continuous presence of thapsigargin caused a small but sustained elevation in [Ca2+]i. These results suggest that dephostatin can mobilize Ca2+ from both a thapsigargin-sensitive and thapsigargin-insensitive intracellular stores in pancreatic acinar cells. In addition, dephostatin can stimulate the release of amylase from pancreatic acinar cells and moreover, reduce the secretory response to CCK-8. The results indicate that dephostatin can release calcium from intracellular calcium pools and consequently induces amylase secretion in pancreatic acinar cells. These effects are likely due to the oxidizing effects of this compound. PMID- 10821433 TI - Butyrate inhibits proliferation-induced proliferating cell nuclear antigen expression (PCNA) in rat vascular smooth muscle cells. AB - Arterial injury-induced vascular smooth muscle cell (VSMC) proliferation in intima is the important etiologic factor in vascular proliferative disorders such as atherosclerosis, hypertension and restenosis after balloon angioplasty. Butyrate, a naturally occurring short chain fatty acid, is produced by bacterial fermentation of dietary fiber and by mammary glands of certain mammals. Studies have shown that butyrate at millimolar concentrations, which are physiological, induces growth arrest, differentiation and apoptosis. We examined the effect of physiological concentrations of butyrate on rat VSMC proliferation and proliferation-induced PCNA expression to determine anti-atherogenic potential of butyrate. Butyrate concentrations, closer to physiological range, exhibited antiproliferative effects on both serum-induced proliferation of serum-starved quiescent VSMCs and actively proliferating non-confluent VSMCs. Treatment of serum-starved quiescent VSMCs with 1-8 mmol/l concentration of butyrate caused a concentration-dependent decrease in serum-induced VSMC proliferation and cell proliferation-associated increase in total cellular proteins and RNA levels. Similarly, exposure of actively growing VSMCs to 5 mmol/l butyrate resulted in the inhibition of cell proliferation and proliferation-induced increase in cellular proteins and RNA levels. Furthermore, cellular morphology was significantly altered. Analysis of cell cycle regulatory proteins indicated that levels of PCNA, an excellent marker for cell proliferation, was significantly altered by butyrate both in actively proliferating and serum-induced quiescent VSMCs. These observations suggest that butyrate exhibits potential antiatherogenic capability by inhibiting VSMC proliferation and proliferation associated increase in PCNA expression and thus merits further investigations regarding therapeutic significance of butyrate in vascular proliferative disorders. PMID- 10821435 TI - Naloxone antagonizes GABA(A)/benzodiazepine receptor function in rat corticohippocampal synaptoneurosomes. AB - Several lines of behavioral and neurochemical evidence indicate GABA(A) antagonistic properties of naloxone. Here, the effects of naloxone on rat brain GABA(A)/benzodiazepine receptor function in vitro were investigated. Naloxone, naltrexone and morphine (10-1,000 microM) reduced GABA-induced (10 microM) 36Cl- uptake in corticohippocampal synaptoneurosomes. Furthermore, the concentration response curve for GABA-induced 36Cl- uptake (GABA 3-100 microM) was shifted to the right both by naloxone and morphine (1,000 microM). Naloxone also reduced the 36Cl- uptake induced by GABA + diazepam (3 microM + 1 microM) but not that induced by amobarbital (500 microM). The naloxone-induced (1,000 microM) reduction of GABA-mediated (10 microM) 36Cl- uptake was reversed by amobarbital (10-1,000 microM) but not by flumazenil (10-1,000 microM) or morphine (0.1-1,000 microM). These results indicate that naloxone, naltrexone and morphine are weak negative modulators of GABA(A)/benzodiazepine receptor function. The naloxone effect most likely does not involve opiate receptors or the benzodiazepine site on GABA(A) receptor complexes. PMID- 10821436 TI - The effect of gender on the melatonin suppression by light: a dose response relationship. AB - It is well known that light is an inhibitor of pineal melatonin secretion in humans. However, the effect of gender on the melatonin suppression by dim and bright light is still controversial. The present study investigated the effect of gender on the suppression of melatonin at five light intensities (0, 200, 500, 1,000, 3,000 lux). Five healthy men and women attended five testing sessions separated by one week. At each session, subjects were exposed to light from midnight to 0100 hours in a sitting position. Blood samples where collected at regular intervals and plasma melatonin concentration was measured using a specific radioimmunoassay. No gender differences were found in melatonin suppression by light at any of the five light intensities (p > 0.1). Furthermore, the mean melatonin suppression by light in both males and females was dose dependent (17% [200 lux], 40% [500 lux], 56% [1,000 lux] and 74% [3,000 lux]). Our findings suggest that melatonin suppression by light in intensity dependent, with no gender differences in light sensitivity. PMID- 10821437 TI - 6-Hydroxydopamine toxicity towards human SH-SY5Y dopaminergic neuroblastoma cells: independent of mitochondrial energy metabolism. AB - 6-Hydroxydopamine (6-OHDA) is widely used to generate animal models of Parkinson's disease. However, little is known about the intracellular events leading to cell death of dopaminergic neurones. Here we correlate indices of energy production and cell viability in human dopaminergic neuroblastoma SH-SY5Y cells after exposure to 6-OHDA. The toxin induces a time and dose-dependent decrease in cell survival with an IC50 value of 25 microM after 24 h. In contrast to the mitochondrial complex I inhibitor 1-methyl-4-phenylpyridinium (MPP+), 6 OHDA-induced reduction of cell viability is not associated with a decrease of intracellular ATP content, intracellular ATP/ADP ratio or NAD+ content. In addition, preventing or forcing glycolysis do not alter 6-OHDA toxicity. The antioxidant D-alpha-tocopherol can attenuate cell death induced by 6-OHDA. These results suggest that cell death induced by 6-OHDA is not due to an inhibition of mitochondrial energy supply, but probably involves production of free radicals. PMID- 10821439 TI - Striatal [123I]beta-CIT SPECT and prefrontal cognitive functions in Parkinson's disease. AB - Twenty non-demented patients with idiopathic Parkinson's disease (PD) underwent single photon emission computed tomography (SPECT) with [123I]beta-CIT to further investigate the contribution of nigrostriatal dysfunction to cognitive and motor deficits. Compared to matched controls PD patients showed normal verbal intelligence, short-term memory and phasic alertness. There were significant (p < 0.05) deficits in tests of verbal working memory (digit ordering, reading span), strategic memory (story recall) and executive functions (card sorting), indicating a "prefrontal" cognitive deficit. Significant (p < 0.05) correlations were observed between dopamine transporter (DAT) density in the putamen and motor deficits as well as between DAT density in both striatal compartments (head of the caudate nucleus and putamen) and prefrontal functioning. Age was a major contributing factor to both cognitive status and nigrostriatal integrity as measured by [123I]beta-CIT SPECT. These results support the view that the striatum is part of a neuronal network that is mediating prefrontal cognitive functions. PMID- 10821438 TI - Differential effects of subchronic treatments with atypical antipsychotic drugs on dopamine D2 and serotonin 5-HT2A receptors in the rat brain. AB - The effects of 3-week treatment with a typical antipsychotic drug chlorpromazine and three atypical antipsychotic drugs (risperidone, olanzapine and perospirone) on the binding to dopamine D2 and serotonin 5-HT2A receptors were examined in the rat stratum and frontal cortex, respectively. Subchronic treatment with chlorpromazine (10 mg/kg) and perospirone (1 mg/kg) significantly increased D2 receptors, while no increase was observed with lower dose of chlorpromazine (5 mg/kg), perospirone (0.1 mg/kg), risperidone (0.25, 0.5 mg/kg) or olanzapine (1, 2 mg/kg). On the other hand, 3-week administration of chlorpromazine (5, 10 mg/kg) and olanzapine (1, 2 mg/kg) significantly decreased 5-HT2A receptors, but risperidone (0.25, 0.5 mg/kg) or perospirone (0.1, 1 mg/kg) had no effect. The measurement of in vivo drug occupation for D2 and 5-HT2A receptors using N ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) suggested that high occupation of 5-HT2A receptors with lower D2 receptor occupancy might be involved in the absence of up-regulation of D2 receptors after subchronic treatment with some atypical antipsychotic drugs. PMID- 10821440 TI - CSF copper concentrations, blood-brain barrier function, and coeruloplasmin synthesis during the treatment of Wilson's disease. AB - During the treatment of four patients with cerebral manifestation of Wilson's disease, we measured the copper concentration in the cerebrospinal fluid (CSF) and serum, the serum coeruloplasmin concentration, the free copper concentration in the serum, and the albumin ratio CSF/serum (AR). These measurements were treated as indicators of the copper-related toxic effects on the brain and the blood-brain barrier (BBB). The half-life of the decrease in the CSF copper concentration during therapy was 23.5 +/- 5.78 months (mean +/- S.E.M.). The therapeutic-target-copper concentration in the CSF (mean normal concentration) is below 20 microg/l. The average length of therapy needed to normalize CSF-copper values in our patients with an average initial value of 76.25 microg/L was 47 month. During the first 10 month of treatment there was an increase in all cases of the measured disturbance in the blood-brain barrier (measured as the ratio of albumin in CSF to albumin in serum, AR). All patients showed an initial worsening of the neurological condition, on average after 1.75 +/- 0.25 months. The maximal rise in AR, from the initial values, was on average 18.4 +/- 5.08%; this maximum was reached after an average of 6.9 +/- 1.5 months. The AR normalized during therapy, indicating a reduction in toxicity in the blood-brain barrier region. The extent of the AR increases in individual patients did not correlate significantly with CSF copper half-life, serum copper half-life, the initial half life of the reduction in the ratio (copper in serum)/(coeruloplasmin in serum), the initial copper concentration in CSF or serum, the initial free copper concentration in serum, or the initial dose of penicillamine (within the first 2 months). We conclude that the normalization of the CSF copper concentration in patients with the cerebral manifestation of Wilson's disease is a slow process, even if therapy is sufficient. The initial worsening of the neurological condition which has often been reported may be reflected in the disturbance of blood-brain barrier function, which we have measured here for the first time (using the parameter of the albumin ratio CSF/serum). Based on repeated measurements of the AR during the course of treatment it seems that the brain toxicity of mobilized copper can be assessed and the therapy adjusted. PMID- 10821441 TI - Glutathione S-transferase polymorphism in Parkinson's disease. AB - Glutathione S-transferases (GSTs) are involved in the detoxification of endogenous or exogenous toxins, which may play a role in the pathogenesis of Parkinson's disease. We genotyped the Glutathione-S-Transferase isoenzymes GSTM1 and GSTT1 by polymerase chain reaction in order to evaluate different gene polymorphisms of these isoenzymes in 149 parkinsonian and 99 control subjects. No differences appeared between both groups regarding the frequencies of the homozygous deletion of GSTM1 (odds ratio 1.021; 95% CI [0.613; 1.699], p < 0.521 Fisher's exact test) and GSTT1 (odds ratio 1.514; 95% CI [0.811; 2.824], p < 0.127). Age of onset of PD did not correlate to GSTM1 and GSTT1 polymorphisms. These results do not support the hypothesis of a possible impact of GSTM1 and GSTT1 detoxification activities in the pathogenesis of Parkinson's disease. PMID- 10821442 TI - Caspase activities and tumor necrosis factor receptor R1 (p55) level are elevated in the substantia nigra from parkinsonian brain. AB - The activities of caspase-1 and caspase-3 were measured by use of fluoropeptides as substrates for the first time in the brain (substantia nigra, caudate nucleus, putamen, cerebellum, and frontal cortex) from control and parkinsonian patients. The activities of caspases in the brain were significantly higher in the substantia nigra from parkinsonian patients than those in the brain from control patients (p < 0.01). However, the activities of caspases in the caudate nucleus, putamen, cerebellum, and frontal cortex showed no significant difference between parkinsonian and control patients. The tumor necrosis factor (TNF) receptor R1 (TNF-R1, p55) level was also elevated in the substantia nigra of the parkinsonian brain in comparison with that of controls (p < 0.05). Since both caspases and TNF R1 may play important roles in apoptotic cell death through TNF-alpha-induced signaling pathway, our present data suggest the presence of a proapoptotic environment in the substantia nigra of parkinsonian brain, probably inducing vulnerability of neurons and glias towards a variety of noxious factors. PMID- 10821443 TI - Tryptophan degradation and immune activation in Alzheimer's disease. AB - Alzheimer's disease (AD) is likely associated with systemic immune activation. During immune response, interferon-gamma stimulates indoleamine 2,3-dioxygenase (IDO) converting tryptophan to N-formylkynurenine followed by kynurenine in an ensuing step. Thus, IDO activity is estimated by the kynurenine per tryptophan quotient (Kyn/Trp). In 21 patients suffering from AD, in 20 controls of similar age, and in 49 blood donors we measured serum tryptophan and kynurenine concentrations by HPLC. Lower tryptophan concentrations were found in elderly control subjects compared to blood donors (62.1 vs. 73.0 microM, p < 0.005). Tryptophan concentrations tended to be still lower in AD patients (54.4 microM, p = 0.07) compared to elderly controls. Enhanced tryptophan degradation in patients was reflected by significantly increased Kyn/Trp (46.1 vs. 34.1 in elderly controls, p < 0.05). Correlations were found in patients between Kyn/Trp and concentrations of soluble immune markers in serum, i.e., neopterin, interleukin-2 receptor and tumor necrosis factor receptor (all p < 0.001). Increased Kyn/Trp was associated with reduced cognitive performance. Tryptophan degradation due to immune activation may exert impact on the pathogenesis of AD. PMID- 10821444 TI - Effects of low-level lead on glycolytic enzymes and pyruvate dehydrogenase of rat brain in vitro: relevance to sporadic Alzheimer's disease? AB - Lead is known to be a potent inhibitor of many enzymes working in the brain, thus possibly inducing functional problems in the brain under pathophysiological conditions. Among such enzymes are those involved in glucose metabolism and energy production. We investigated the inhibitory effects of low-level lead on brain hexokinase (HK), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), pyruvate kinase (PK) and pyruvate dehydrogenase complex (PDHc) with rat brain homogenate. PDHc was distinctively inhibited when low-dose lead acetate was added last of all (IC50 = 5 microM) to the reaction mixture. The other enzymes were completely resistant to 5 microM of lead acetate. When the homogenate was preincubated with lead acetate HK was dramatically inhibited by low-level lead acetate (1-5 microM), in a manner dependent on both preincubation time and lead concentration. However, the inhibitory effect was abolished by coincubation with its substrates, glucose or ATP. The results suggest that exposure to low levels of lead may increase the risk of cerebral hypometabolism caused by direct inhibition of specific glucose-utilizing enzymes. In this context, lead might be regarded as a risk factor in the abnormal glucose metabolism seen in some kinds of neurodegenerative disorders such as sporadic Alzheimer's disease. PMID- 10821445 TI - Long-term oscillation of corticosterone following intermittent cocaine. AB - We have shown that repeated administration of cocaine, as well as other drugs and nondrug stressors, can induce alternating increases and decreases in several neurotransmitter and endocrine endpoints, which we call oscillation. Oscillation studies have typically used 3-4 pretreatments with cocaine or other agents, raising the question of whether oscillation lasts beyond this point. Using plasma corticosterone as our endpoint measure, we therefore inquired whether oscillation would persist across eight administrations of cocaine over a 28-day period. We report oscillation of corticosterone levels persisting across all eight cocaine groups. Our data also indicate that the degree of oscillation increases with the intertreatment interval. PMID- 10821446 TI - Modulation of 5-HT1A receptor mediated response by fluoxetine in rat brain. AB - Radioligand binding studies were done to investigate the effect of chronic administration of fluoxetine on 5-HT1 receptor mediated response to adenylate cyclase (AC) in rat brain. Our studies revealed a significant decrease in the densities of 5-HT1 and 5-HT1A receptor sites in cortex and hippocampus of rat brain after chronic administration of fluoxetine (10 mg/Kg body wt.). However there was no significant change in the affinity of [3H]5-HT and [3H]DPAT for 5 HT1 and 5-HT1A receptor sites, respectively. However, in striatum, along with a significant (75%) downregulation of 5-HT1 sites, the affinity of [3H]5-HT to these sites was increased, as revealed by decrease in Kd (0.50 +/- 0.08 nM). Displacement studies showed that fluoxetine has higher affinity for 5-HT1A receptors with a Ki value of 14.0 +/- 2.8 nM, than 5-HT1 sites. No significant change was observed in basal AC activity in any region after fluoxetine exposure. However, in cortex of experimental rats the 5-HT stimulated AC activity was significantly increased (16.03 +/- 0.97 pmoles/mg protein; p < 0.01), when compared to 5-HT stimulated AC activity (12.98 +/- 0.78 pmoles/mg protein) in control rats. The increase in 5-HT stimulated AC activity in cortex may be due to the significant downregulation of 5-HT1A sites in cortex after fluoxetine exposure as these sites are negatively coupled to AC. The observed significant decrease in 5-HT1 sites with concomitant increase in 5-HT stimulated AC activity, after fluoxetine treatment, suggests that fluoxetine, which has high affinity for these sites, acts by modulating the 5-HT1A receptor mediated response in brain. PMID- 10821447 TI - A survey of surgical audit in Australia: whither clinical governance? AB - A postal survey was undertaken to determine the extent of surgical audit in Australian public hospitals in the context of clinical governance. Participants were 88 surgical units from 74 Australian public hospitals in 1998 with more than 70 beds. The main outcome measures were performance of audit, methods used, frequency, documentation and perceived adequacy of resources and comparison of performance with Royal Australasian College of Surgeons requirements for re certification. The main findings were that most units performed audit, but over two-thirds declared that they had insufficient resources to conduct audit according to College requirements, while a substantial proportion did not comply with other published guidelines for the conduct of audit meetings. The conclusion from this survey is that surgical audit will be unable to play its part in quality initiatives inherent in the clinical governance concept unless appropriately resourced. PMID- 10821448 TI - Standardising surveillance of nosocomial infections: the HISS program. Hospital Infection Standardised Surveillance. AB - Standardised surveillance of nosocomial infections in Australia had not been addressed until June 1998 when the New South Wales Health Department funded the development and implementation of the first standardised surveillance system for hospital infection: the Hospital Infection Standardised Surveillance program (HISS). The introduction of a standardised surveillance system needs to balance the requirements of a Health Department and the needs of hospitals. The Health Department requires data to develop aggregated rates for the setting of thresholds for all nosocomial infections while hospitals require rates to reflect the quality of clinical care and provide data for evidence-based infection control practices. The Hospital Infection Epidemiology and Surveillance (HIES) Unit has attempted to balance these requirements using a 'sentinel surveillance' approach with standardised definitions and methodology. The HISS program utilizes eICAT software modified for its standardised requirements of data collection. To date, 10 hospitals surveyed sentinel multiple resistant organisms (MRO), eight also elected sentinel surgical procedures (SSP) and intravascular device-related bacteraemia (IVDRB) modules, and two the seasonal respiratory syncytial (RSV) and rota-virus modules in paediatric patients. The surgical site infection rates in three commonly monitored SSP were 1.8% (95% confidence interval (CI) 0.7-3.9%) for coronary artery bypass (CABG), 3.3% (95% CI 1.4-6.8%) lower segment Caesarean section (LSCS) and 7.7% (95% CI 3.4-14.6%) colorectal surgery. The rate of IVDRB was 4.7 per 1000 central venous catheter days (95% CI 2.2-8.6) and 1.1 per 1000 peripheral line-days (95% CI 0.1-3.9). Methicillin resistant Staphylococcus aureus (MRSA) accounted for 99% of all new infections diagnosed with an endemic MRO. PMID- 10821449 TI - Optimising care of acute myocardial infarction: results of a regional quality improvement project. AB - The effects of a quality improvement intervention were evaluated in a before after time-series study of 649 consecutive patients suffering acute myocardial infarction (AMI) in the West Moreton Health District over 2.5 years from March 1996 through to August 1998. After a 6-month baseline period, clinical practice guidelines were issued followed by sequential feedback to providers of clinical indicator data over a 1-year period. Resultant changes in practice were then evaluated during a 12-month post-intervention period. The proportion of eligible patients receiving early thrombolysis, lipid-lowering drugs and cardiac rehabilitation increased, respectively, from 30.8 to 70.0% (P = 0.001), from 23.4 to 56.4% (P = 0.003), and from 23.6 to 54.3% (P = 0.003). The in-hospital death rate, incidence of postinfarct angina and mean length of stay decreased, respectively, from 15.8 to 8.6% (P = 0.02), from 30.1 to 14.3% (P < 0.001), and from 7.4 to 6.3 days (P = 0.001). Despite the absence of control groups, the present study suggested that clinical guidelines combined with feedback of clinical indicators were useful in improving quality of care. PMID- 10821450 TI - Patterns of use, attitudes and expectations of mental health staff regarding computers. AB - There is limited information on the views of mental health staff about computers. The present study aimed to ascertain the patterns of use, attitudes and expectations of staff from a comprehensive mental health service regarding computers before and after the purchase of new equipment and training. A questionnaire was sent to staff of the Central Sydney Mental Health Services working in sites targeted for new computer equipment and training. Most respondents, especially those with computer experience or who had worked in mental health for less than 5 years, viewed computers favourably. At the same time, half the respondents did not feel they had sufficient access to a computer at work and the vast majority had not received training as intended. Commitments to provide computer equipment and training must be followed through, otherwise staff may feel disenchanted and computers may be regarded less favourably. A measure of the positive views about computer use ('positivity index') developed during the course of the current study may have wider applicability. PMID- 10821451 TI - Use of pathology services in re-engineered clinical pathways. AB - A significant proportion of pathology tests ordered in hospital are unnecessary. Specific measures targeting the increasing appropriateness of pathology service use have been shown to decrease overall ordering of laboratory tests. However, it is not clear whether general programmes to improve quality of care will have any impact on the use of pathology services. Use of pathology services was compared within two separate prospective controlled clinical trials of re-engineered clinical pathways for both elective (surgical) patients and acute unplanned (medical) admissions. Trial One was a controlled trial of a re-engineered surgical service. Booked patients in the treatment group were admitted on the day of surgery, care was guided by a clinical pathway, and patients were discharged early with domiciliary post-acute care. Controls were admitted on the day before surgery, treated according to usual practice and discharged according to surgeons' preferences. In Trial Two, acute medical patients admitted to hospital through the Emergency Department (ED) were randomised into a treatment (Hospital in the Home) or a control (inpatient) care pathway. In both studies, patients on the re-engineered clinical pathways were well matched demographically and clinically. Health outcomes and satisfaction ratings were comparable. Seventy per cent fewer laboratory tests were ordered in the elective surgery intervention group (P < 0.0001), while the treatment group of the acute medical patients had 25% fewer tests ordered (P = 0.0133). Pooled results also showed a significantly lower rate of test ordering (P < 0.001) for the treatment group (Mann-Whitney U Wilcoxon ranked sum test). The findings of these audits of controlled, prospective trials suggested overuse of laboratory tests in New South Wales public hospitals, and that savings can be generated by using clinical pathways and applying clinical criteria to the ordering of tests without adversely affecting health outcomes. PMID- 10821452 TI - Reflections from the chair of the National Expert Advisory Group: implementing safety and quality enhancement in health care. PMID- 10821453 TI - What is the quality of our quality managers? Is it time for quality managers in Australia to be certified? PMID- 10821454 TI - Postgraduate courses in quality improvement: achievements and future directions. AB - Since 1996, the Centre for Clinical Epidemiology and Biostatistics at the University of Newcastle has offered courses in Quality Improvement in Health Care. The courses are offered at Graduate Certificate, Graduate Diploma and Masters levels, principally by distance learning. They offer training in research methods as well as the concepts and application of quality improvement. Feedback from students, many of whom are quality coordinators in Australian health care facilities, has been positive. Enrolments are taken mid-year as well as at the beginning of the academic year. PMID- 10821455 TI - The ACHS Care Evaluation Program: a decade of achievement. Australian Council on Healthcare Standards. AB - In 1989 the Australian Council on Healthcare Standards (ACHS) embarked on a programme to develop acute health care clinical indicators in conjunction with the Australian medical colleges. Through a carefully structured stepwise process this collaboration established a 'World first' in 1993 with the introduction of the first set of indicators into the ACHS Accreditation programme. The programme remains unique in the formal involvement of providers in the development process and in the scope of the clinical areas covered in acute health care. From the year 2000 there will be 18 sets (and over 200 indicators) from which health care organisations (HCOs) can choose to monitor the major services they provide. There remains no compulsion to address a specific number of indicators. The growth of the programme has been considerable with more than half of the nations' acute HCOs reporting their clinical indicator data (twice yearly) and it provides a reflection of the care given for the majority of patient separations in acute care. This reporting process allows HCOs to receive feedback on the aggregate results together with comparative peer group information for each indicator they address. In addition to numerous publications in peer reviewed journals an annual aggregate report, 'the Measurement of Care in Australian Hospitals' is published. It reports both qualitative and quantitative data on all indicator sets for the preceding year. Validity of the indicators is strengthened each year with a review process and reliability and reproducibility of the data can now be demonstrated. The clinical response to the indicators has been overwhelming and there is now documented evidence of numerous actions taken by HCOs to improve both the processes and the outcomes of patient care. The nation wide database can be expected to reflect trends in care over the next few years. The process of indicator refinement, however, will continue and it is likely that a reduction in the total number of indicators will occur with a core group of the more 'robust' indicators remaining. Further directions in indicator development are likely to be in the area of multidisciplinary care and in the assessment of longer-term outcomes. In addition to measures of the quality of care, hopefully, in time, health care providers will also take part in the establishment of measures of the appropriateness of that care. PMID- 10821456 TI - Quality use of allopurinol in the elderly. AB - Allupurinol is a commonly prescribed drug. However, the use of this drug is not based on evidence and guidelines. We audited Allopurinol prescriptions in patients aged 65 years and over in a teaching hospital over 22 weeks. In 47% of patients the dose was higher than recommended and in 40% it was lower. Quality use of medications is an important issue to maintain quality of life in the elderly. PMID- 10821457 TI - The development of a national agreed minimum diabetes dataset for New Zealand. AB - The development of a minimum diabetes dataset (MDD) for monitoring diabetes in New Zealand was intended to facilitate diabetes quality initiatives. Existing published datasets were reviewed and a draft MDD for New Zealand was distributed to all 147 specialist, general practice and relevant community groups. Data definitions were either identical or compatible with other datasets and dataset items included if there were at least six supportive replies. All groups were followed up by letter and telephone. A total of 26 (18%) replies were received. Comments were reviewed and the recommended MDD finalised. There was agreement that this dataset would be embedded into the software of at least three commercially available patient management systems. In conclusion, developling an agreed national MDD was difficult, in spite of its potential utility for local, regional and national collation of diabetes data allowing those involved to generate a picture of diabetes and its outcomes. PMID- 10821458 TI - Medication containers and the law. PMID- 10821459 TI - Autoimmune and antitumor consequences of antibodies against antigens shared by normal and malignant tissues. AB - There is now a considerable body of information documenting the autoimmune consequences of antibodies induced by growing malignancies, or by passively administered and actively induced antibodies, in cancer patients against antigens shared by normal and malignant tissues. This provides a rich source of information addressing the consequences of autoantibodies against a range of antigens. Antibodies against cell-surface or intracellular antigens in the central nervous system (CNS) or on epithelial surfaces of normal tissues do not generally result in autoimmunity, but the same types and titers of antibodies against cell surface antigens in the subepidermal skin, peripheral nerves, blood, or vascular sites such as the spleen and bone marrow readily induce autoimmunity. The blood brain barrier of the CNS and apical antigen expression and the basement membrane in epithelial tissues, may protect these sites from antibody induced damage. Cancer cells, however, are protected by neither unidirectional antigen expression nor basement membranes. Vaccine induced antibodies against a variety of cancer cell surface antigens have been associated with prevention of tumor recurrence in preclinical models and in vaccinated cancer patients, in the absence of demonstrable autoimmunity. This forms the basis for a series of ongoing Phase III trials with single or polyvalent antigen cancer vaccines designed for optimal antibody induction. PMID- 10821460 TI - The comparison of the efficacy and safety of intravenous versus subcutaneous immunoglobulin replacement therapy. AB - To compare the efficacy of immunoglobulin replacement therapy given intravenously versus subcutaneously to prevent infections in patients with primary antibody deficiency syndromes, an international, multicenter, open label, crossover study was designed. Forty patients were randomized to receive either subcutaneous or intravenous immunoglobulin replacement therapy for 1 year. In the second year, patients were switched to the alternative treatment, enabling patients to act as their own controls. Equivalent doses were given by both routes. Ethical approval was obtained from the review boards of the hospitals in which the patients were seen and written consent obtained from each patient. Patients with a primary antibody deficiency syndrome, either common variable immunodeficiency or IgG subclass deficiency or specific antibody deficiency, who required immunoglobulin replacement therapy were included in the study. Patients were excluded if they had significant thrombocytopenia (defined as platelets less than 50 x 10(9)/liter), had high levels of anti-IgA antibodies (defined as greater than 1:8192), or had severe adverse reactions to a blood product within the last 2 years. The primary end point was the number of infections and their severity (moderate and major) during the two treatment periods. Secondary end points were adverse reactions, length of infections, days lost from school or work due to infections, and acceptability of treatment regimens to the patients. Based on the assumption that it was difficult to prove equivalence of therapies statistically in crossover studies, an arbitrary number of 40 patients was selected on the basis that this might be achievable in 2 years. There are no significant differences in efficacy or adverse reaction rates between immunoglobulin replacement therapy given subcutaneously or intravenously. PMID- 10821461 TI - Soluble antiapoptotic molecules and immune activation in chronic heart failure and unstable angina pectoris. AB - Programmed myocyte cell death and activation of the immune system have been shown to occur in patients with congestive heart failure. Besides, unstable angina episodes are likely to be associated with immune activation. Our aim was to evaluate the role of changes in circulating levels of soluble Fas (sFas), suggestive of an enhanced inhibitory response to ongoing apoptosis, and soluble IL2 receptor (sIL2-R), indicative of T-lymphocyte activation, in chronic heart failure and unstable angina pectoris. Thirty patients affected by chronic heart failure (20 idiopathic and 10 ischemic cardiomyopathy) and 13 patients with unstable angina were evaluated. Twenty healthy individuals matched for age and gender were used as controls. A complete biochemical determination of indexes of myocardial damage including cardiac troponin I (cTnI) and creatine kinase (MB/CK) was performed. The results demonstrated that mean levels of sFas and sIL2-R were significantly increased in patients affected by chronic heart failure and unstable angina and were not associated with changes in renal function or with serum levels of cTnI. Highest values of sFas were found in NYHA class IV patients (IV NYHA class = 7.39 +/- 0.52 vs. controls = 1.34 +/- 0.12 ng/ml; P < 0.01) and more elevated in idiopathic than in ischemic cardiomyopathy (3.64 +/- 0.40 vs. 1.82 +/- 0.37 ng/ml; P < 0.01). Moreover, in chronic heart failure patients sFas and ejection fraction were negatively correlated (P = 0.01), whereas sFas and sIL2-R were positively correlated (P < 0.01). In unstable angina patients too, sFas and sIL2-R appeared to be correlated (P = 0.03); whereas sFas (angina group = 3.18 +/- 0.39 vs. controls = 1.34 +/- 0.12 ng/ml; P < 0.01) and sIL2-R (angina group = 0.46 +/- 0.11 vs. controls = 0.00 UI/ml; P < 0.01) were higher in angina group than in controls. In most of the cases, the increase of sFas was associated with comparable changes in sIL2-R serum levels, indicating that the activation of Fas system is strictly associated with autoimmune-inflammatory reactions. This phenomenon, both in chronic heart failure and in unstable angina, occurs in the absence of biochemical evidences of myocardial damage and seems to parallel the activation of T cell. Soluble Fas could have a role in sustaining inflammatory response and in prolonging the detrimental effects correlated with it in chronic heart failure and angina pectoris. PMID- 10821462 TI - Il-4 and IFN-gamma mRNA induction in human peripheral lymphocytes specific for beta-lactam antibiotics in immediate or delayed hypersensitivity reactions. AB - Beta-lactam antibiotics elicit CD4+ and CD8+ T-cell-mediated immune responses that play a central role in allergic reactions. However, the involvement of a type 1- (Th1 or Tc1) or a type 2-like (Th2 or Tc2) differentiation in drug allergy remains unclear. We investigated the expression of interleukin 4 (IL-4) and interferon gamma (IFN-gamma) mRNA by quantitative reverse transcription and polymerase chain reaction (RT-PCR) in patient-derived peripheral blood lymphocytes following specific in vitro stimulation. Samples were collected from a total of 19 patients who had developed immediate or delayed clinical manifestations of hypersensitivity to beta-lactam and from 11 control subjects. Peripheral blood mononuclear cells (PBMCs) were stimulated with either free antibiotics or antibiotic-human serum albumin (HSA) conjugates. Specific induction of IFN-gamma mRNA expression was observed in 11 of 11 allergic patients with immediate reactions, in 6 of 8 patients with delayed reactions, and in 4 of 11 control subjects. IL-4 mRNA expression was induced in 5 of 11 allergic individuals with immediate reactions but in none of the 8 patients with delayed responses and none of the 11 control subjects. IL-4 mRNA expression was only induced following activation with free drugs, while IFN-gamma mRNA expression was predominantly induced in CD4+ T cells following stimulation with HSA-conjugated drugs. Immediate-type hypersensitivity to beta-lactams was not associated with a pure type 2-like response when PBMCs were specifically stimulated in vitro: Some patients with well-documented history of beta-lactam-induced immediate allergic reaction showed a high IFN-gamma response. Contact dermatitis involved Tc1 and Th1 cells and other delayed hypersensitivity reactions to beta-lactams were restricted to Th1 responses. PMID- 10821463 TI - Elevated plasma levels of 90K (Mac-2 BP) immunostimulatory glycoprotein in HIV-1 infected children. AB - 90K is a secreted serum glycoprotein with immune stimulatory activity. In this study, 90K plasma levels were determined by an enzyme-linked immunosorbent assay in 18 HIV-1-infected children and 10 uninfected control children. 90K levels in HIV-1-infected children (median, 12.5 microg/ml) were higher than in HIV-1 uninfected control group (6.3 microg/ml; P < 0.05). 90K levels of HIV-1-infected children classified as stage B and C (median, 15.0 microg/ml and 22.7 microg/ml, respectively) were higher compared to children with stage A disease (median, 7.0 microg/ml; P < 0.05). A positive correlation (r = 0.5; P < 0.05) was found between 90K levels and HIV-1 RNA levels in 137 plasma samples of 18 HIV-1 infected children collected during a period of 1 year. No correlation was found between 90K levels and CD4 cell counts. These results suggest that 90K plasma levels may represent a novel marker of disease progression in HIV-1-infected children. PMID- 10821464 TI - Urine free light chains in SLE: clonal markers of B-cell activity and potential link to in vivo secreted Ig. AB - As a marker of in vivo B-cell activity, urine levels of free light chain (FLC) were measured twice weekly by radioimmunoassay (RIA) and correlated with disease activity over periods of 5-10 months in seven patients with systemic lupus erythematosus (SLE). In addition, RIA-measured urine albumin was used to track glomerular injury, and alpha1-microglobulin (alpha1-M) levels, 28- to 32-kDa protein, provided control measurements on excretion of low-molecular-weight proteins. As controls, urine FLC levels were obtained from healthy normals and in subjects with acute pharyngitis, sickle-cell anemia, and acute sepsis or pneumonia. The control results showed that with acute sepsis/pneumonia had marked increases in urine FLC, while pharyngitis and sickle-cell controls had normal FLC levels. In SLE, active patients receiving intravenous cyclophosphamide and high dose steroids exhibited highly increased urine FLC that fluctuated widely during therapy and fell to normal range levels with disease remission. During active SLE, urine albumin often was increased, while alpha1-M levels remained in normal range. In contrast to the increased FLC of active disease, inactive patients on low-dose maintenance therapy had predominantly normal FLC levels throughout the collection period. These results support our hypothesis that longitudinal levels of urine FLC can be used to track disease-related B-cell activity in SLE. Furthermore, we suggest that the urine FLC of active SLE would share LC idiotype with the clonal associated in vivo secreted Ig, and thus permit the identification of these antibodies that are targeted to the culprit immunogen(s) responsible for the pathogenesis of SLE. PMID- 10821465 TI - Vaccination responses to capsular polysaccharides of Neisseria meningitidis and Haemophilus influenzae type b in two C2-deficient sisters: alternative pathway mediated bacterial killing and evidence for a novel type of blocking IgG. AB - Meningitis caused by Neisseria meningitidis serogroup W-135 was diagnosed in a 14 year-old girl with a history of neonatal septicemia and meningitis caused by group B streptococci type III. C2 deficiency type I was found in the patient and her healthy sister. Both sisters were vaccinated with tetravalent meningococcal vaccine and a conjugate Haemophilus influenzae type b vaccine. Three main points emerged from the analysis. First, vaccination resulted in serum bactericidal responses demonstrating anticapsular antibody-mediated recruitment of the alternative pathway. Second, addition of C2 to prevaccination sera produced bactericidal activity in the absence of anticapsular antibodies, which suggested that the bactericidal action of antibodies to subcapsular antigens detected in the sera might strictly depend on the classical pathway. A third point concerned a previously unrecognized type of blocking activity. Thus, postvaccination sera of the healthy sister contained IgG that inhibited killing of serogroup W-135 in C2-deficient serum, and the deposition of C3 on enzyme-linked immunosorbent assay plates coated with purified W-135 polysaccharide. Our findings suggested blocking to be serogroup-specific and dependent on early classical pathway components. Retained opsonic activity probably supported post-vaccination immunity despite blocking of the bactericidal activity. The demonstration of functional vaccination responses with recruitment of alternative pathway-mediated defense should encourage further trial of capsular vaccines in classical pathway deficiency states. PMID- 10821466 TI - Mucosal immune response in the ear, nose and throat. PMID- 10821467 TI - Bacterial otitis media: pathogenetic considerations. AB - A variety of exciting and important new observations regarding the pathogenesis of nontypable H. influenzae infection have been made in the past decade. The interactions between mucin and OMPs show a high degree of specificity. Multiple adhesins have been identified on the bacterial surface. Colonization of the upper respiratory tract is a dynamic process. Immunodominant, antigenically heterogeneous OMPs are the targets of strain-specific immune responses, accounting in part for the recurrent nature of OM in otitis-prone children. The LOS of nontypable H. influenzae displays a remarkable degree of antigenic and phase variation and may be involved in molecular mimicry of host antigens. Finally nontypable H. influenzae not only lives on the mucosal surface but also clearly has been demonstrated to enter epithelial cells and remain viable in intracellular and intercellular locations in the human upper respiratory tract. These areas of investigation have important implications in understanding the pathogenesis of OM. Elucidating mechanisms of pathogenesis will be important in guiding development of novel ways to prevent OM. PMID- 10821468 TI - Role of viruses in the pathogenesis of acute otitis media. AB - To date there is ample evidence suggesting a crucial role for respiratory viruses in the pathogenesis of AOM. Respiratory viral infection appears to initiate the cascade of events that finally leads to development of AOM (Fig. 1). The pathogenesis of AOM is complicated, involving a network of factors, some probably not yet identified, which affect each other in a time-dependent manner. Increased knowledge of the detailed mechanisms of viral infection, the host inflammatory response during URI and the interaction between viruses and bacteria could lead to major advances in the prevention of AOM. PMID- 10821469 TI - Viral and bacterial interaction in acute otitis media. PMID- 10821470 TI - Clinical epidemiology of otitis media. PMID- 10821471 TI - Clinical course, complications and sequelae of acute otitis media. PMID- 10821472 TI - Increasing antibiotic resistance among otitis media pathogens and their susceptibility to oral agents based on pharmacodynamic parameters. PMID- 10821473 TI - Clinical significance of antibiotic resistance in acute otitis media and implication of antibiotic treatment on carriage and spread of resistant organisms. PMID- 10821474 TI - Prevention of otitis media in children with live attenuated influenza vaccine given intranasally. PMID- 10821475 TI - Potential of bacterial vaccines in the prevention of acute otitis media. PMID- 10821476 TI - Conjugate pneumococcal vaccine and antibiotic-resistant Streptococcus pneumoniae: herd immunity and reduction of otitis morbidity. PMID- 10821477 TI - Nonimmune strategies for prevention of otitis media. PMID- 10821478 TI - Hypoxia, clonal selection, and the role of HIF-1 in tumor progression. AB - Tumor progression occurs as a result of the clonal selection of cells in which somatic mutations have activated oncogenes or inactivated tumor suppressor genes leading to increased proliferation and/or survival within the hypoxic tumor microenvironment. Hypoxia-inducible factor 1 (HIF-1) is a transcription factor that mediates adaptive responses to reduced O2 availability, including angiogenesis and glycolysis. Expression of the O2-regulated HIF-1alpha subunit and HIF-1 transcriptional activity are increased dramatically in hypoxic cells. Recent studies indicate that many common tumor-specific genetic alterations also lead to increased HIF-1alpha expression and/or activity. Thus, genetic and physiologic alterations within tumors act synergistically to increase HIF-1 transcriptional activity, which appears to play a critical role in the development of invasive and metastatic properties that define the lethal cancer phenotype. PMID- 10821479 TI - Geminiviruses: models for plant DNA replication, transcription, and cell cycle regulation. AB - Geminiviruses have small, single-stranded DNA genomes that replicate through double-stranded intermediates in the nuclei of infected plant cells. Viral double stranded DNA also assembles into minichromosomes and is transcribed in infected cells. Geminiviruses encode only a few proteins for their replication and transcription and rely on host enzymes for these processes. However, most plant cells, which have exited the cell cycle and undergone differentiation, do not contain the replicative enzymes necessary for viral DNA synthesis. To overcome this barrier, geminiviruses induce the accumulation of DNA replication machinery in mature plant cells, most likely by modifying cell cycle and transcriptional controls. In animals, several DNA viruses depend on host replication and transcription machinery and can alter their hosts to create an environment that facilitates efficient viral replication. Analysis of these viruses and their proteins has contributed significantly to our understanding of DNA replication, transcription, and cell cycle regulation in mammalian cells. Geminiviruses have the same potential for plant systems. Plants offer many advantages for these types of studies, including ease of transformation, well-defined cell populations and developmental programs, and greater tolerance of cell cycle perturbation and polyploidy. Our knowledge of the molecular and cellular events that mediate geminivirus infection has increased significantly during recent years. The goal of this review is to summarize recent research addressing geminivirus DNA replication and its integration with transcriptional and cell cycle regulatory processes. PMID- 10821480 TI - Health-related quality of life among adults with arthritis--behavioral risk factor surveillance system, 11 states, 1996-1998. AB - Arthritis and other rheumatic conditions are the leading cause of disability in the United States, affecting 43 million persons in 1998 and-with the aging of the "baby boom" generation-are projected to affect an estimated 60 million by 2020. In 1992, these conditions cost $65 billion in medical care and lost productivity and were associated with 744,000 hospitalizations and 44 million ambulatory-care visits in 1997. Because arthritis and other rheumatic conditions seldom cause death but have a substantial impact on health, health-related quality of life (HRQOL) measures are better indicators of their impact than related mortality rates. This report examines data from 11 states that included an arthritis module in the 1996-1998 Behavioral Risk Factor Surveillance System (BRFSS); findings indicate that persons with arthritis have worse HRQOL than persons without arthritis, regardless of sex, age, or education level. PMID- 10821481 TI - Morbidity and mortality associated with Hurricane Floyd--North Carolina, September-October 1999. AB - On September 16, 1999, Hurricane Floyd, a storm extending 300 miles with sustained winds of 96-110 miles per hour, made landfall in North Carolina, dropping up to 20 inches of rain in eastern regions of the state. Rain from Hurricane Floyd, combined with rains from Hurricane Dennis beginning on August 30 and Hurricane Irene on October 17, caused extensive flooding along the Neuse, Tar, Roanoke, Lumbar, and Cape Fear rivers, affecting an estimated 2.1 million persons. This report presents data about injuries, illnesses, and deaths during and following Hurricane Floyd in North Carolina and identifies the leading cause of death as drowning involving occupants of motor vehicles trapped in flood waters. PMID- 10821482 TI - Surveillance for possible estuary-associated syndrome--six states, 1998-1999. AB - Pfiesteria piscicida (Pp) is an alga that has been associated with fish kills in estuaries (where fresh water mixes with salty seawater) along the eastern seaboard and possibly with human health effects. Since June 1, 1998, surveillance for possible estuary-associated syndrome (PEAS), including possible Pp-related human illness, has been conducted in Delaware, Florida, Maryland, North Carolina, South Carolina, and Virginia. This report summarizes passive surveillance for PEAS during June 1, 1998-December 31, 1999, which indicated no persons had illnesses that met PEAS criteria. PMID- 10821483 TI - Update: influenza activity--United States and worldwide, 1999-2000 season, and composition of the 2000-01 influenza vaccine. AB - Influenza A (H3N2) viruses were the predominant viruses isolated in the United States and worldwide during 1999-2000. This was the third consecutive year that influenza A/Sydney/05/97-like (H3N2) viruses were the most prevalent viruses isolated in the United States. Influenza activity in the United States was similar to the previous two seasons, although mortality measurements attributed to pneumonia and influenza (P&I) were unusually high. Overall, the 1999-2000 influenza vaccine was well matched to circulating influenza viruses. The 2000-01 influenza season will be the first for which influenza vaccination is recommended for all persons aged > or =50 years. This report summarizes surveillance for influenza in the United States and worldwide during the 1999-2000 influenza season, describes the composition of the 2000-01 influenza vaccine, and highlights changes in the recommendations for prevention and control of influenza. PMID- 10821484 TI - Angiogenesis as a predictor of clinical outcome in cancer patients. PMID- 10821485 TI - Neuroendocrine expression in metastatic prostate cancer: evaluation of high throughput tissue microarrays to detect heterogeneous protein expression. AB - The theory that poorly differentiated prostate carcinoma develops a neuroendocrine (NE) phenotype is controversial. Supportive data is variable with NE expression being observed in anywhere from 5% to 83% of prostate cancers. These percentages are derived from standard immunohistochemistry studies, which make no attempt to quantify the results. High-density tissue microarrays (TMAs), represent a novel method for evaluating up to 1000 tissue samples with a 0.6 mm diameter on a single glass slide. This high throughput technology for screening antibodies, however, requires validation to determine if TMAs are useful in evaluating heterogeneously expressed proteins such as the NE markers chromogranin A (CGA) and synaptophysin (SYN). This study compares results from standard slides to TMAs in 50 primary and metastatic prostate tumors taken from 12 rapid autopsies from men with hormone refractory prostate cancer. One hundred standard and 2 TMA slides were immunostained for CGA and SYN. Using standard slides, focal NE expression was seen in 1/12 primary prostate tumors. Overall, 13/100 (13%) standard slides showed focal NE expression for both primary and metastatic prostate tumors; NE expression was observed in 4/12 autopsy cases (33%) when all tumor sites per case were considered. 458 tissue elements (tumor and normal) were arrayed into one paraffin block. Seventy-three percent (332/458) of the elements placed into the TMA were confirmed histologically to represent tumor. Seventy five percent (250/332) and 66% (218/332) could be evaluated for CGA and SYN expression, respectively. Six of the metastatic tumors expressed CGA and SYN or 2.4% (6/250; 95% CI = 0.9% to 5.2%) and 2.3% (6/218; 95% CI = 0.8% to 5.3%), respectively. In conclusion, only focal NE expression was observed by both methods (eg, standard and TMA slides). The focal expression in these advanced prostate tumors was unexpected given data from prostate tumor cell lines and animal models suggesting that progression to the NE phenotype parallels tumor progression. This study also supports the use of high density TMAs to screen for protein expression, even when expression is focal. PMID- 10821486 TI - NCAM (neural cell adhesion molecules) expression in malignant mesotheliomas. AB - Neural cell adhesion molecules (NCAM) are adhesion molecules expressed by neural and neuroendocrine tumors and a few biphasic tumors such as synovialosarcomas and breast phyllode tumors. To investigate NCAM expression in mesotheliomas, we studied 26 cases of epithelioid (n = 12), biphasic (n = 11), and sarcomatoid (n = 3) malignant mesotheliomas (MM), in comparison with normal mesothelium, and 50 primary non-small cell lung carcinomas (NSCLC) (25 adenocarcinomas [ADC] and 25 squamous cell carcinomas [SCC]), using electron microscopy as a gold standard for recognition of MM. NCAM reactivity using 123C3 antibody was compared with that of NE markers such as chromogranin A and synaptophysin. Although normal mesothelium remains negative, NCAM was expressed in 19 of 26 MM (73%) with a membranous staining on frozen or paraffin sections. In 6 of 12 epithelioid MM, the tumor cells expressed NCAM, whereas in 5 cases stromal fibroblasts showed a strong but focal staining. In 11 biphasic MM, 4 presented an NCAM reactivity of both epithelioid and spindle cell components, whereas in 7, only fusiform component was NCAM positive. Two of 3 sarcomatoid MM showed an NCAM expression. Chromogranin expression was never seen, whereas synaptophysin was noticed in 2 cases. No case of NSCLC showed membranous 123C3 staining, whereas 2 ADC weakly expressed synaptophysin. We conclude that NCAM expression in MM is reminiscent of its expression in mesoderm during fetal life and consistent with that reported in other biphasic tumors. These data show that NCAM expression occurs in 73% of MM, highly exceeding that observed in lung cancer. PMID- 10821487 TI - Clonality assessment of lymphoproliferative disorders by multiparameter flow cytometry of paraffin-embedded tissue: an additional diagnostic tool in surgical pathology. AB - A major drawback of immunohistochemical detection of monoclonality in B-cell lymphoproliferative disorders is the lack of contrast between surface immunoglobulin staining and extracellular immunoglobulin staining. To bypass this drawback, immunophenotyping of single-cell suspensions by flow cytometry is commonly used. Although the expression of immunoglobulin light chain subtype can be quantified rapidly and reliably, the technique is hampered by the requirement of fresh unfixed material. We applied a recently developed technique for the isolation of single cells from formalin-fixed, paraffin-embedded material to measure clonality in B-cell lymphoproliferative disorders (lymphoid tissue (n = 10) and non-Hodgkin's B-cell lymphoma (n = 10). Immunocytochemistry indicated that common cell surface markers as well as the immunoglobulin light chains could be detected in the cell suspensions derived from archival material. In addition, the technique also allowed combined high-resolution DNA flow cytometric analysis. To investigate the effect of formalin fixation on cross-linking of extracellular immunoglobulins to lymphocytes, a double-immunostaining experiment for both light chain immunoglobulins (kappa and lambda) was performed. This experiment showed that this cross-linking was minimal (less than 2%). All cases of reactive lymphoid hyperplasia were DNA diploid and showed a polyclonal expression of immunoglobulin light chains. In contrast, in 9 of 10 non-Hodgkin's B-cell lymphomas, monoclonality was established on the basis of light chain expression, whereas only 6 of 9 cases were conclusive by immunohistochemistry. Four of the 9 cases were DNA aneuploid. One case did not show light chain expression at all by both techniques. However, this case could be classified as malignant by flow cytometric analysis because of the DNA-aneuploid nature of the B-cell subpopulation. The average S-phase fraction (SPF) of the B cells in the reactive lymphoid tissues was 3.5%. The mean SPF values for B cells in DNA-diploid cases of lymphomas was 3.0%, whereas the mean SPF of B cells in DNA-aneuploid cases was 6.1%. The presented technique is superior to immunohistochemistry for the detection of monoclonality in B-cell lymphoproliferative disorders and therefore provides a powerful tool to support the diagnosis of malignant lymphoma in routinely processed archival samples of lymphoid tissues. PMID- 10821488 TI - Galectin-3 expression in various thyroid neoplasms and its possible role in metastasis formation. AB - Galectin-3 is a member of the beta-galactoside-binding protein family that plays an important role in cell-cell adhesion and in cell-matrix interaction. We have examined the expression of galectin-3 in normal, adenomatous, and malignant thyroid tissues and also in metastatic lesions. Galectin-3 was rarely expressed in normal thyroid tissue but was abundant in the cytoplasm of the neoplastic lesions. Among neoplastic lesions, galectin-3 was expressed to a greater extent in follicular carcinomas than in follicular adenomas and was present in greater amounts in papillary carcinomas than in follicular adenomas or carcinomas. Primary lesions of papillary carcinoma with metastasis contained significantly higher concentrations of galectin-3 than tumors of this type without metastases. However, the expression of galectin-3 was significantly decreased in metastatic lesions in the lymph nodes compared with their primary lesions. From these results, we assumed that galectin-3 works in different ways at different stages of thyroid neoplasm proliferation. Among primary tumors, galectin-3 expression is significantly different in 3 histological types. However, the continuity of progression among these tumors is not yet proven. In later stages, decreased expression of galectin-3 may aid the release of cancer cells from the primary lesions for invasion and metastasis. PMID- 10821489 TI - Decreased expression of Fas (CD95/APO1) associated with goblet cell metaplasia in Barrett's esophagus. AB - Fas ligand (FasL) has been shown to induce apoptosis in cells expressing its receptor Fas. We have recently shown that Fas ligand is overexpressed in all cases of Barrett's metaplasia (BM) with dysplasia and esophageal adenocarcinomas, and in a few cases of BM negative with dysplasia. The aim of this work was to determine the status of Fas expression in BM with and without dysplasia or carcinoma. Formalin-fixed and paraffin-embedded tissue sections from esophageal biopsies and esophagectomy specimens with BM, with and without dysplasia and carcinoma, were immunostained for Fas and FasL using the immunoperoxidase technique. The percentage of positive cells in each case was evaluated and compared with the degree of dysplasia. When Fas expression was assessed in glands with goblet cell metaplasia, Fas immunoreactivity was either undetected or present in less than 10% of the cells in 85% of the cases, and only 1 (4%) of the 28 cases examined showed Fas immunoreactivity in more than 25% of the cells. When we compared Fas expression in goblet cell-containing glands with glands of gastric cardia phenotype, we found that in the 26 cases of BM with or without dysplasia Fas was completely undetectable in goblet cell-containing glands in 15 (58%) of the cases but was undetectable in only 3 (12%) of the glands with gastric cardia phenotype (P = .002). Fas is usually undetectable or is expressed at a low level in BM with or without dysplasia or carcinoma. Fas expression in goblet cell-containing glands is less frequent than in glands with gastric cardia phenotype in the same specimens. BM with dysplasia or carcinoma overexpress FasL, so decreased Fas expression may protect BM with dysplasia and carcinoma from self destruction while allowing them to evade immune surveillance. PMID- 10821490 TI - Increased angiogenesis in papillary thyroid carcinoma but lack of prognostic importance. AB - Angiogenesis is important for tumor growth and progress, and the intratumor density of microvessels is a significant prognostic factor in many tumor types. The role in thyroid cancer has not been well studied, and we therefore examined a series of 128 papillary carcinomas with respect to microvessel density (MVD) and patient survival. Follow-up was obtained for all cases (median, 145 months). We found a mean MVD of 216 per mm2 (range, 35-751), and there was an average of 3.14 times more vessels in the tumors, when compared with surrounding non-neoplastic thyroid tissue. MVD was inversely related to age, tumor diameter, histological grade, and primary tumor extent. Furthermore, increasing MVD tended to be associated with improved survival (P = .056). In conclusion, our data indicate that angiogenesis is important for the development and maintenance of papillary thyroid carcinomas, although it was not identified as a prognostic factor. PMID- 10821492 TI - Malignant cell detection by fluorescence in situ hybridization (FISH) in effusions from patients with carcinoma. AB - Cytological diagnosis of malignant cells in effusions is hampered by difficulties in the differentiation from reactive mesothelial cells. Because interphase cytogenetics by fluorescence in situ hybridization (FISH) might complement cytological evaluation, we determined the power of tumor cell detection using FISH and cytology in 201 effusions from patients with advanced cancer. Furthermore, 9 primary breast tumors were FISH-karyotyped, and chromosomal aberrations were compared with those of corresponding metastatic effusion cells. By using centromeric probes representing chromosomes 7, 8, 11, 12, 17, and 18, a rate of malignancy-associated aneusomy combined for the 6 chromosomes was detected in an overall of 44.8% of effusion specimens (range, 31.8% to 39.3% for the individual chromosome), comparable to cytology (43.3%). The combination of just 2 FISH probes (namely, representing chromosome pairs 8/11 and 8/17) was almost equally efficient in the identification of aneusomy. Approximately one fourth of the cytologically negative effusions were FISH positive and vice versa. From the initially FISH-negative effusions, 18.9% could be subsequently classified positive with dual-color FISH by visualization of intranuclear chromosomal complexity in rare aneuploid cells. Thus, "overall FISH analysis," including dual-color evaluation, identified tumor cells in significantly more effusions (55.2%, P = .001) than conventional cytology, implying greater sensitivity. Finally, our finding that numerical aberration patterns in primary breast tumors and corresponding metastatic effusions are comparable indicates that FISH examination of primary tumors will indicate the centromeric probe(s) best suited for an efficient search for metastasis in the individual case. PMID- 10821493 TI - Endometriosis-associated intestinal tumors: a clinical and pathological study of 6 cases with a review of the literature. AB - This clinicopathologic study of primary Mullerian tumors of the bowel arising in foci of endometriosis is based on six new cases and an analysis of 17 previously reported cases. Varieties of Mullerian tumors occur in the bowel; the most common types are endometrioid carcinoma, followed by various mixed Mullerian tumors and stromal sarcomas. Seventy-eight percent develop in the rectosigmoid colon, the remaining in the cecum or ileum. Those in the latter area tend to be sarcomas or mixed Mullerian tumors. Certain architectural growth characteristics, derived from precursor endometriosis, are common to most endometriosis-associated intestinal tumors (EAITs). Seventy percent of EAITs occur in the outer bowel wall. Transmural tumors tend to form luminal polyps and assume an hourglass shape. Metachronous or synchronous Mullerian tumors occur in 39% of cases. Seventy percent of women with EAITs are in their mid 30s to early 50s. Common presenting symptoms are abdominal or pelvic pain, melena, and an abdominal or pelvic mass. Documented in 26% of patients is a history of prolonged unopposed estrogen therapy. Only 28.5% of cases die of their tumors, but follow-up is less than 5 years in all but 2 patients. PMID- 10821491 TI - Tissue factor expression and angiogenesis in human prostate carcinoma. AB - In tumors, the switch to the angiogenic phenotype is thought to be controlled by a balance of positive and negative angiogenic factors. Tissue factor (TF) produced by tumor cells has been implicated in the regulation of this "angiogenic switch" through its ability to concurrently induce the expression of angiogenic molecules such as vascular endothelial cell growth factor (VEGF), while inhibiting the expression of anti-angiogenic molecules such as thrombospondin 2. We have examined TF expression and its relationship to angiogenesis and tumor progression in human prostate carcinomas. Most of the prostate carcinoma specimens examined (73%; n = 67) express high levels of TF. Immunohistochemical analysis localized TF expression to the epithelial cells of malignant glands. TF expression was significantly correlated with tumor angiogenesis as measured by the microvessel density (MVD). In addition, TF expression was correlated with the preoperative PSA level, a strong predictor of recurrence in prostate carcinomas. Our findings show that TF expression by the malignant glands in prostate cancer is common and suggest a role for this molecule in regulating prostate cancer progression and angiogenesis. PMID- 10821494 TI - Pattern of distribution of intraductal and infiltrating ductal carcinoma: a three dimensional study using serial coronal giant sections of the breast. AB - The purpose of this study was to establish the 3-dimensional (3D) structure of the breast tissue and to study the distribution and relationship between the intraductal and infiltrating components of ductal carcinoma and other proliferative epithelial lesions of the breast. Thirty mastectomy specimens with infiltrating carcinoma less than 3.0 cm in diameter were serially cut in the coronal plane. Each giant section was divided into small sections for routine processing. Using Photoshop (Adobe) and PowerPoint (Microsoft) software programs, the routinely stained sections were scanned and assembled to reestablish complete giant sections of the breast and subsequently the 3D structure. Intraductal and infiltrating ductal carcinomas, epithelial hyperplasia with atypia, and marked epithelial hyperplasia without atypia were mostly confined to a single duct (27 cases), resulting in an increase in size of the involved breast segment. Three remaining cases included a case of Paget's disease with tumor appearing to spread from one duct system to another system through the epidermis and two cases with multiple separate foci of carcinomas located in different quadrants and accompanied by ductal spread in different lactiferous ducts. Both intraductal and infiltrating carcinomas were often located in the superficial segments (near the subcutaneous tissue) (28 cases). The infiltrating components were often located adjacent to area of pure intraductal carcinoma and were often peripheral (nearer the chest wall than the nipple). Intraductal carcinomas showed a "fanned out" pattern of distribution, frequently extended toward the nipple (with involvement of the nipple or subareolar tissue in 7 cases), and occasionally were seen in the breast tissue peripheral to the infiltrating carcinoma. Multiple ducts with intraductal carcinoma could be seen to be connected with each other with serial sections. However, in at least 6 cases, foci of intraductal carcinomas were separated from each other by segments of duct with benign epithelium. Breast carcinoma often arise from the breast segment close to the subcutaneous tissue. Infiltrating carcinoma lesser than 3.0 cm in diameter is usually located adjacent to the area of pure intraductal. The pattern of spread of intraductal carcinoma has a pyramid-like shape, with the summit toward and occasionally extending up to the nipple. These findings should be considered in the surgical strategy for segmental resections of breast carcinomas. PMID- 10821495 TI - Thioredoxin, a putative oncogene product, is overexpressed in gastric carcinoma and associated with increased proliferation and increased cell survival. AB - Human thioredoxin is a putative oncogene that may confer both a growth and survival advantage to tumor cells. Overexpressed thioredoxin mRNA has been found in both primary human lung and colorectal cancers. To determine the intratumor distribution and amount of thioredoxin protein in human primary carcinomas, we developed an immunohistochemical assay for thioredoxin in paraffin-embedded tissue. We then studied 10 patients with primary high-risk gastric carcinoma. To further relate thioredoxin protein overexpression to cell death and survival, we used a paraffin-based in situ end-labeling (ISEL) assay. To delineate proliferation, we used the nuclear proliferation antigen detected by Ki-67. In this survey, we found that thioredoxin was localized to tumor cells and overexpressed compared with normal gastric mucosa in 8 of 10 gastric carcinomas. The thioredoxin was found at high levels in 5 of the 8 overexpressing carcinomas. The overexpression of thioredoxin was typically found in both a nuclear and cytoplasmic location in the neoplastic cells. There was a significant positive correlation (P = .0061) with cancer cell proliferation measured by Ki-67. There was a significant negative correlation (P = .0001) with DNA damage measured by the ISEL assay, suggesting decreased apoptosis and increased carcinoma cell survival. Thus, human primary gastric tumors that are highly expressive of thioredoxin have both a higher proliferative rate and a higher survival rate than tumors that do not express thioredoxin. With these newly developed assays in hand, it is now feasible to question whether this thioredoxin-related combined growth and survival advantage translates into poor clinical outcome. PMID- 10821496 TI - The expression of Ep-CAM (17-1A) in squamous cell cancers of the lung. AB - Immunotherapy trials using monoclonal antibodies 323/A3 and 17-1A that recognize Ep-CAM, including trials focused on cancer of the lung, currently are underway. Nevertheless, there have been few comprehensive evaluations of the expression of Ep-CAM in specific types of neoplastic processes, including cancer of the lung. The current study of 60 human subjects with squamous cell cancer (SCC) of the lung, selected at random, was undertaken (1) to examine the expression of Ep-CAM in SCC and associated uninvolved bronchial mucosa, bronchial epithelial hyperplasia, and dysplasia, and (2) to correlate the results with established prognostic indicators and survival of patients. In both the uninvolved bronchial mucosa and epithelial hyperplasia, the expression of Ep-CAM in luminal cells was significantly higher compared with its expression in the matched basal cells (P = .003, P < .0001, respectively). When Ep-CAM scores of basal and luminal cells present in uninvolved bronchial mucosa and epithelial hyperplasia were combined, we observed a statistically significant stepwise increase in Ep-CAM expression from uninvolved bronchial mucosa to epithelial hyperplasia to SCC, suggesting its involvement in malignant transformation of SCC. The expression of Ep-CAM was significantly higher in poorly to moderately differentiated SCC compared with well-differentiated SCC (P = .04). An increase in the expression of Ep-CAM with increasing size or local extent of the primary tumor approached statistical significance (P = .09). The expression of Ep-CAM increased significantly with increasing involvement of regional lymph nodes (P = .02). Similarly, the expression of Ep-CAM increased with the increasing TNM stages (P = .04). Kaplan Meier Survival analysis using the same categorizations showed that increasing tumor size, nodal status, and stage were significantly associated with poor patient survival (P = .04, .01, .01, respectively). There was, however, no statistically significant association between patient survival and staining intensity of carcinomas for Ep-CAM. We conclude that expression of Ep-CAM increased during the progression of SCC of the lung and, therefore, may play a role in the carcinogenesis of this disease. PMID- 10821497 TI - The cutaneous manifestations of human parvovirus B19 infection. AB - The prototypical cutaneous manifestations of human parvovirus B19 (B19) infection include a petechial eruption in a glove and stocking distribution, reticular truncal erythema, and the "slapped cheek" sign. An association with connective tissue disease (CTD) stigmata has recently been made. The clinical and dermatopathologic findings in 14 patients whose skin lesions were accompanied by serological evidence of B19 infection or documentation of B19 genome in lesional skin are presented. The authors encountered skin biopsy specimens from 14 patients who presented with skin eruptions accompanied by clinical signs or serology suggestive of antecedent B19 infection. Clinical findings were correlated to the light microscopic appearance of the lesions and the presence of B19 genome in lesional skin. The study group comprised 9 women, 3 men, and 2 boys. Eruptions characteristic of fifth disease, including the slapped cheek sign, reticulated truncal erythema, and acral petechiae, were present in 3 patients, 1 of whom later developed granuloma annulare. The other patients had atypical clinical presentations comprising an asymptomatic papular eruption (2), an eruption clinically resembling Sweet's syndrome (3), myopathic dermatomyositis (DM) (2), lupus erythematosus (LE)-like syndromes (2), and lower-extremity palpable purpura (2). Skin biopsy specimens in 12 cases showed interstitial histiocytic infiltrates with piecemeal fragmentation of collagen and a mononuclear cell-predominant vascular injury pattern. Other features included an interface dermatitis, eczematous alterations, and papillary dermal edema. Lesions with features of DM or LE also showed mesenchymal mucinosis, whereas a biopsied lesion of palpable purpura showed leukocytoclastic vasculitis (LCV). Immunofluorescent testing showed a positive lupus band test (LBT) with epidermal IgG and C5b-9 decoration in 1 patient with a systemic LE-like illness, whereas the DM patients had negative LBTs and vascular C5b-9 deposition typical for DM. Skin biopsy specimens from 11 patients, including those whose presentations resembled LE and DM, were positive for B19 genome. The dermatopathology of B19 infection suggests tissue injury mediated by delayed-type hypersensitivity, by antibody-dependent cellular immunity directed at microbial antigenic targets in the epidermis and endothelium, and by circulating immune complexes in the setting of LCV. These mechanisms appear to generate a clinical and histopathological picture that recapitulates that of CTD. PMID- 10821498 TI - Molecular evidence that the stromal and epithelial cells in pleomorphic adenomas of salivary gland arise from the same origin: clonal analysis using human androgen receptor gene (HUMARA) assay. AB - Salivary gland pleomorphic adenomas are characterized by a biphasic growth of "epithelial" and "stromal" regions. The "epithelial" region is a compactly organized mixture of both luminal and nonluminal cells, whereas the stromal region is composed predominantly of the nonluminal cells. Using the polymerase chain reaction (PCR)-based HUMARA assay on DNA from formalin-fixed, paraffin embedded tissues from pleomorphic adnomas of female patients, we intend to clarify the clonal relation between the luminal and nonluminal cells and the clonal nature of the morphologically diverse nonluminal cells in this tumor. HUMARA, the human androgen receptor gene, is located on the X chromosome and contains a segment of polymorphic CAG tandem repeats in exon 1. Several methylation-sensitive HhaI restriction sites are located 5' to these CAG repeats. It is an ideal tool to study clonality of female tissues by examining the methylation pattern. Of the 13 cases analyzed, 3 were homozygous at the HUMARA locus and therefore noninformative. The remaining 10 cases were informative. All 10 cases showed a monoclonal pattern in the stromal area, indicating that the morphologically diverse nonluminal cells are monoclonal. Eight of the 10 cases showed monoclonality in the "epithelial" areas, suggesting a common clonality between luminal and nonluminal cells. Of the remaining 2 samples, 1 was polyclonal for the "epithelial" region, and the other was not amplifiable. Our data provide the first molecular evidence that the luminal and nonluminal cells in pleomorphic adenomas arise from the same clone in most cases, and the morphologically diverse nonluminal cells are monoclonal. PMID- 10821499 TI - 10q23.3 loss of heterozygosity is higher in lymph node-positive (pT2-3,N+) versus lymph node-negative (pT2-3,N0) prostate cancer. AB - Loss of heterozygosity (LOH) in the region of 10q23.3 has been associated with multiple tumors, including glioblastoma multiforme, melanoma, endometrial carcinoma, and prostate carcinoma. The tumor suppressor gene, PTEN/MMAC1, is also located in this region, and, in addition to other tumor types (eg, glioblastoma multiforme, endometrial, and melanoma), PTEN/MMAC1 mutations have been found in prostate cancer cell lines, xenografts, and hormone refractory prostate cancer tissue specimens. The aim of this study was to evaluate LOH at 10q23.3 as a marker of cancer progression in node-positive prostate cancer. Genetic alterations in the region of 10q23.3 were assessed in 23 node-positive (pT2-3, N+) and 44 node-negative prostate (pT2-3, N0) cancers with D10S532, D10S1687, D10S541, and D10S583 flanking polymorphic genetic markers; PTENCA, a genetic marker within PTEN/MMAC1, was also tested. Using DNA from paired normal and microdissected tumor samples, LOH at microsatellite loci was determined after polymerase chain reaction amplification. LOH in at least 1 marker was identified in 14% (6 of 44) of lymph node-negative and 43% (10 of 23) of lymph node-positive prostate cancers (chi-square test, P = .007). This increase in genetic alterations in node-positive prostate cancer suggests that 10q23.3 is a marker for metastatic progression. PMID- 10821500 TI - Trefoil factor expression in normal and diseased human salivary glands. AB - Trefoil factors are wound-healing peptides important in protection and healing of the human gastrointestinal tract. Their potential for therapy of gastrointestinal ulcers has been established. This study investigated the hypothesis that trefoil factors are also present in human salivary gland. Tissues from surgical biopsy specimens were collected fresh into ice and stored in liquid nitrogen. Breast, stomach, and colon constituted positive controls. Trefoil factor mRNAs were detected by reverse transcription polymerase chain reaction (RT-PCR) or by in situ hybridization (ISH) with formalin-fixed, paraffin-embedded sections. Amplified DNA fragments were ligated into pGEM-T Easy vector and used to transform competent Escherichia coli JM109, allowing sequencing to confirm identity of cloned fragments. Generation of amplifiable cDNA was confirmed using primers specific to the ubiquitously expressed abl gene. By RT-PCR, TFF1 (pS2) mRNA was detected in 14 of 15 glands, TFF3 (hITF) mRNA in 13, and TFF2 (hSP) in only 1 gland. ISH of 15 glands (7 of which had been studied by RT-PCR) showed the same pattern of expression and indicated that TFF1 mRNA was usually expressed at low levels by a few mucous cells, whereas TFF3 was produced abundantly by most mucous cells. There was no difference in patterns of expression comparing parotid, submandibular, and minor mucous glands. Nor was there an obvious relationship between trefoil factor expression and pathology, but those glands not expressing TFF1 or TFF3 had evidence of chronic inflammation or atrophy. Trefoil factors are likely to be important in healing, predisposition to, and therapy of, oral diseases. PMID- 10821501 TI - Malignant pigmented clear cell epithelioid tumor of the kidney: clear cell ("sugar") tumor versus malignant melanoma. AB - A 73-year-old woman presented with an hemorrhagic kidney tumor initially interpreted as a renal cell carcinoma (RCC). A retroperitoneal recurrence infiltrating the duodenal wall was made up of clear cells, some of which contained Fontana-Masson positive pigment, immunopositive for HMB-45, S-100 protein, actin, and vimentin. The same immunohistochemical profile was retrospectively reproduced in the kidney tumor, where melanosomes were also found ultrastructurally. Lipomatous differentiation was not observed. There was no history of malignant melanoma (MM), or stigmata of tuberous sclerosis. The patient died of disease 5 years after the initial diagnosis. This neoplasm can be considered as a malignant, pigmented, clear-cell epithelioid variant of angiomyolipoma, or "sugar" tumor of the kidney, with the peculiarity of having a previously unreported component of pigmented cells visible on light microscopy. This finding raises the possibility that the exceptional cases of MM reported in renal parenchyma may be pigmented variants of epithelioid angiomyolipoma rather than true MM. PMID- 10821502 TI - Epithelioid sarcoma and its variants. PMID- 10821503 TI - Assessment of proliferative activity in breast cancer: MIB-1 immunohistochemistry versus mitotic figure count. PMID- 10821504 TI - Principles of functional magnetic resonance imaging: application to auditory neuroscience. AB - Functional imaging based on magnetic resonance methods is a new research frontier for exploring a wide range of central nervous system (CNS) functions, including information processing in sensory, motor, cognitive, and linguistic systems. Being able to localize and study human brain function in vivo, in relatively high resolution and in a noninvasive manner, makes this a technique of unparalleled importance. In order to appreciate and fully understand this area of investigation, a tutorial covering basic aspects of this methodology is presented. We introduce functional magnetic resonance imaging (fMRI) by providing an overview of the studies of different sensory systems in response to modality specific stimuli, followed by an outline of other areas that have potential clinical relevance to the medical, cognitive, and communicative sciences. The discussion then focuses on the basic principles of magnetic resonance methods including magnetic resonance imaging, MR spectroscopy, fMRI, and the potential role that MR technology may play in understanding a wide range of auditory functions within the CNS, including tinnitus-related activity. Because the content of the material found herein might be unfamiliar to some, we provide a broad range of background and review articles to serve as a technical resource. PMID- 10821505 TI - Toward a more ecologically valid measure of speech understanding in background noise. AB - In an attempt to develop a more ecologically valid measure of speech understanding in a background of competing speech, we constructed a quasidichotic procedure based on the monitoring of continuous speech from loudspeakers placed directly to the listener's right and left sides. The listener responded to the presence of incongruous or anomalous words imbedded within the context of two children's fairy tales. Attention was directed either to the right or to the left side in blocks of 25 utterances. Within each block, there were target (anomalous) and nontarget (nonanomalous) words. Responses to target words were analyzed separately for attend-right and attend-left conditions. Our purpose was twofold: (1) to evaluate the feasibility of such an approach for obtaining electrophysiologic performance measures in the sound field and (2) to gather normative interaural symmetry data for the new technique in young adults with normal hearing. Event-related potentials to target and nontarget words at 30 electrode sites were obtained in 20 right-handed young adults with normal hearing. Waveforms and associated topographic maps were characterized by a slight negativity in the region of 400 msec (N400) and robust positivity in the region of 900 msec (P900). Norms for interaural symmetry of the P900 event-related potential in young adults were derived. PMID- 10821506 TI - Longitudinal investigation of hearing disorders in children with congenital cytomegalovirus. AB - This investigation consisted of a longitudinal study of the effects of congenital cytomegalovirus (CMV) infection on hearing sensitivity in 860 children with documented asymptomatic or symptomatic congenital CMV infection. Of the 651 children with asymptomatic CMV infection, 48 (7.4%) developed sensorineural hearing loss (SNHL), compared to 85 (40.7%) of the children with symptomatic CMV infection. Children in both groups experienced latent effects consisting of delayed onset of loss, threshold fluctuations, and/or progressive loss of hearing. It can be concluded that congenital CMV infection is a leading cause of SNHL in children. The late onset and progression of loss necessitates continued monitoring of hearing sensitivity in this population. PMID- 10821507 TI - Different associations of blood lead, meso 2,3-dimercaptosuccinic acid (DMSA) chelatable lead, and tibial lead levels with blood pressure in 543 former organolead manufacturing workers. AB - In this study, the authors' objective was to determine the influence of blood lead, meso 2,3-dimercaptosuccinic acid (DMSA)-chelatable lead, and tibial lead on systolic and diastolic blood pressures and on hypertension in 543 former organolead manufacturing workers. All workers had past exposure to inorganic and organic lead. The authors used linear regression to model systolic and diastolic blood pressure separately, and logistic regression was used for the modeling of hypertension status (i.e., systolic blood pressure > 160 mm Hg, diastolic blood pressure > or =96 mm Hg, or current use of antihypertensive medications). Blood lead, DMSA-chelatable lead, and tibial lead levels had means (standard deviations appear within parentheses) of 4.6 microg/dl (2.6 microg/dl), 19.3 microg (17.2 microg), and 14.4 microg/g (9.3 microg/g), respectively. The authors adjusted for covariates, and they found that blood lead was a predictor of (1) both systolic and diastolic blood pressures and (2) hypertension status in men < 58 y of age. DMSA-chelatable lead and tibial lead were not associated with any of the blood pressure measures. Systolic blood pressure was elevated by blood lead levels as low as 5 microg/dl. We speculate that lead may have a transient influence on blood pressure that is related to target dose levels obtained once release of lead from body stores has occurred. PMID- 10821508 TI - Impact of soil and dust lead on children's blood lead in contaminated areas of Sweden. AB - The impact of lead in soil and dust on blood lead concentrations in young children (i.e., 1-5 y of age, N = 202) and the risk of health effects were investigated in an urban and a mining area of Sweden. Blood, soil, and indoor dust, as well as information on lead-exposure factors, were collected. The blood lead concentrations (total range = 9-77 microg/l) the authors measured indicated a low risk for lead-induced health effects. Lead in soil (i.e., < 10-5,000 microg/g) and in dust (i.e., < 1-316 microg/g) had little effect on blood lead concentrations, given the present conditions and present concentration range- especially in the mining area. Urban children had significantly higher blood lead concentrations than children in the mining area, despite higher concentrations of lead in soil in the mining area. In the urban children, blood lead concentrations were influenced by parental smoking and lead in dust at day-care centers. PMID- 10821509 TI - Renal effects of cadmium intake of a Japanese general population in two areas unpolluted by cadmium. AB - Renal effects of cadmium exposure (i.e., in food products) on people living in two areas unpolluted by cadmium in Japan were investigated. The population comprised 875 inhabitants (i.e., 346 males and 529 females) and 635 inhabitants (i.e., 222 males and 413 females), all of whom in each area were 50+ y of age. The authors used urinary cadmium concentration as an indicator of internal dose, and total urinary protein, beta2-microglobulin, and N-acetyl-beta-glucosaminidase were indicators of renal dysfunction. The authors used multiple-regression and logistic-regression analyses to study the relationship between urinary cadmium excretion and the above indicators of renal dysfunction. In the two geographic areas, both analyses revealed that urinary cadmium concentrations were associated significantly with indicators of renal dysfunction. The results suggest that renal dysfunction is related significantly with environmental cadmium exposure in areas of Japan that are not polluted by cadmium. PMID- 10821510 TI - Prevalence and features of advanced asbestosis (ILO profusion scores above 2/2). International Labour Office. AB - In this study, the author addressed the following question: Do workers with advanced asbestosis have a restrictive pulmonary physiology, and, alternately, do those who have restrictive physiological tests have advanced asbestosis? One group was identified by obvious radiographic measurements, and the other group was defined via physiologic measurements. Total lung capacity, vital capacity, and flows were measured in 12,856 men exposed to asbestos, of whom 3,445 had radiographic signs of asbestosis, as defined by the International Labour Office criteria. Radiographically advanced asbestosis-International Labour Office criteria profusion greater than 2/2 was present in 85 (2.5%) of men. An additional 52 men had physiologically restrictive disease. The author, who compared pulmonary flows and volumes of these two groups, used mean percentage predicted, adjusted for height, age, and duration of cigarette smoking. Men with radiographically advanced asbestosis had normal total lung capacity (i.e., 105.5% predicted), reduced forced vital capacities (i.e., 82.7% predicted), air trapping (i.e., residual volume/total lung capacity increased to 54.4%), and reduced flows (i.e., forced expiratory flow [FEF25-75] = 60.6% predicted, forced expiratory volume in 1 s = 78.0% predicted, and forced expiratory volume in 1 s/forced vital capacity = 65.5%). In contrast, men selected from the same exposed population for restrictive disease (i.e., reduced total lung capacity [72.6% predicted] and forced vital capacity [61.5% predicted]) also had airflow obstruction (i.e., forced expiratory volume in 1 s/forced vital capacity of 74.5% predicted) and air trapping (i.e., residual volume/total lung capacity of 46.7%). Only half of these men had asbestosis--and it was of minimal severity. In summary, advanced asbestosis was characterized by airway obstruction and air trapping, both of which reduced vital capacity but not total lung capacity; therefore, it was not a restrictive disease. In contrast, restrictive disease was rare and was associated with minimal asbestosis. PMID- 10821512 TI - Air pollution and daily mortality in Shenyang, China. AB - The authors analyzed daily mortality data in Shenyang, China, for calendar year 1992 to identify possible associations with ambient sulfur dioxide and total suspended particulates. Both total suspended particulate concentrations (mean = 430 microg/m3, maximum = 1,141 microg/m3) and sulfur dioxide concentrations (mean 197 = microg/m3, maximum = 659 microg/m3) far exceeded the World Health Organization's recommended criteria. An average of 45.5 persons died each day. The lagged moving averages of air-pollution levels, calculated as the mean of the nonmissing air-pollution levels of the concurrent and 3 preceding days, were used for all analyses. Locally weighted regression analysis, including temperature, humidity, day of week, and a time variable, showed a positive association between daily mortality and both total suspended particulates and sulfur dioxide. When the authors included total suspended particulates and sulfur dioxide separately in the model, both were highly significant predictors of daily mortality. The risk of all-cause mortality increased by an estimated 1.7% and 2.4% with a 100 microg/m3 concomitant increase in total suspended particulate and sulfur dioxide, respectively. When the authors analyzed mortality separately by cause of death, the association with total suspended particulates was significant for cardiovascular disease (2.1%), but not statistically significant for chronic obstructive pulmonary diseases (2.6%). In contrast, the association with sulfur dioxide was significant for chronic obstructive pulmonary diseases (7.4%), but not for cardiovascular disease (1.8%). The mortality from cancer was not associated significantly with total suspended particles or with sulfur dioxide. The correlation between sulfur dioxide and total suspended particulates was high (correlation coefficient = .66). When the authors included sulfur dioxide and total suspended particulates simultaneously in the model, the association between total suspended particulates and mortality from all causes and cardiovascular diseases remained significant. Sulfur dioxide was associated significantly with increased mortality from chronic obstructive pulmonary diseases and other causes. The results of the current study reveal increased mortality associated with both total suspended particulates and sulfur dioxide. PMID- 10821511 TI - Effects of lead and noise exposures on hearing ability. AB - Lead and noise, via different mechanisms, may damage hearing ability, and, in some cases, cause severe and irreversible damage. To explore possible independent and synergistic effects of lead and noise on auditory function, the authors conducted a cross-sectional study in two lead-battery manufacturing factories. Lead and noise were the two most common sources of occupational exposures in the factories. Blood lead level, ambient lead concentration, noise exposure level, and hearing thresholds of 339 lead-battery workers-including clerical and managerial staffs-were measured. The authors obtained demographics and working histories via an interview-based structured questionnaire. A total of 220 lead battery workers were exposed to high levels of lead and noise; their average blood lead concentration was 56.9 microg/dl, and their average noise exposure level was 86.0 dBALeq. Multivariate analysis, in which possible risk factors of hearing ability were considered, demonstrated a significant correlation between a high, long-term lead exposure index (defined by duration of employment and ambient lead concentration) and decreased hearing ability. In contrast, such a correlation between short-term lead exposure (defined by blood lead level) and hearing ability was not significant. Furthermore, neither noise exposure level alone nor the interaction between noise exposure level and short- or long-term lead exposure was correlated significantly with hearing ability. The present study raises an important, but typically ignored, issue: lead exposure might precipitate a more severe auditory than noise-exposure effect. The preservation of workers' hearing ability requires that preventive measures be taken against noise exposure, which is as essential as measures taken against lead exposure. PMID- 10821513 TI - Effect of the Hanshin-Awaji earthquake on posttraumatic stress, lifestyle changes, and cortisol levels of victims. AB - In 1995, the Hanshin-Awaji area in Japan was damaged severely by a major earthquake. In this study, the authors administered questionnaires and obtained blood samples to analyze the relationships among lifestyle, psychological stress, and plasma cortisol levels of victims. The authors questioned 107 male inhabitants of Awaji Island about their lifestyles before and after the earthquake, presence of any symptoms of posttraumatic stress disorder, and demographic variables. Plasma cortisol levels were determined with enzyme immunoassay. Cortisol level was correlated strongly with change in lifestyle. The highest cortisol levels were found in the group characterized by a high posttraumatic stress disorder score and by a very profound lifestyle change. This group also contained the highest percentage of subjects who had poor health. In summary, the psychological stress induced by the Hanshin-Awaji earthquake was associated with mean cortisol level; however, this relationship was affected by adjustment of lifestyle. PMID- 10821514 TI - Effects of air pollution on children's respiratory health in three Chinese cities. AB - During the winter of 1988-1989, parents of 2,789 elementary-school students completed standardized questionnaires. The students were 5-14 y of age and were from three urban districts and one suburban district of three large Chinese cities. The 4-y average ambient levels of total suspended particles in the three cities differed greatly during the period 1985-1988: Lanzhou, 1,067 microg/m3; urban Wuhan, 406 microg/m3; Guangzhou, 296 microg/m3; and suburban Wuhan, 191 microg/m3. The authors constructed unconditional logistic-regression models to calculate odds ratios and 95% confidence intervals for prevalences of several respiratory symptoms and illnesses, adjusted for district, use of coal in the home, and parental smoking status. There was a positive and significant association between total suspended particle levels and the adjusted odds ratios for cough, phlegm, hospitalization for diseases, and pneumonia. This association was derived from only the 1,784 urban children and, therefore, the authors were unable to extrapolate it to the suburban children. The results also indicated that parental smoking status was associated with cough and phlegm, and use of coal in the home was associated only with cough prevalence (alpha = 0.05). PMID- 10821515 TI - Variation in blood lead and hematocrit levels during pregnancy in a socioeconomically disadvantaged population. AB - Lead is a long-recognized human toxicant that crosses the placenta. Fetal sensitivity to environmental agents can vary with stage of development; therefore, how maternal blood lead levels change during pregnancy and how fetal exposure is influenced provide useful knowledge. In this study, the authors describe longitudinal changes in blood lead levels during the course of pregnancy in a sample of socioeconomically disadvantaged pregnant women. The women were recruited early in pregnancy when they sought care at one of two obstetrics clinics in Albany, New York. Maternal blood lead levels changed between the 1st and 2nd trimesters, from 1.99 microg/dl to 1.69 microg/dl (hematocrit corrected, 1.70-1.62); between the 2nd and 3rd trimester from 1.78 microg/dl to 1.86 microg/dl (hematocrit corrected, 1.65-1.72); and between 3rd trimester and delivery from 1.80 microg/dl to 2.17 microg/dl (hematocrit corrected, 1.70-1.86). These changes were statistically significant and were corrected for secular trends. The rate of change per day in lead levels averaged -36.6% from trimester 1 to trimester 2, 18.3% from trimester 2 to trimester 3, and -40.8% from trimester 3 to delivery. The patterns in our study were consistent with the patterns reported in a few other longitudinal studies of change in lead level during pregnancy. Findings reveal significant associations between maternal blood lead levels and both hematocrit and trimester of pregnancy. Clinicians who interpret test results should take into account the dynamics of these variables when determining appropriate care for both mother and neonate. PMID- 10821516 TI - Nutrition: the integrative science. AB - 1. Nutrition is a vitally important part of the production environment but also interacts with many other disciplines. 2. Modern breeding methods involving selection for a wider range of characteristics are likely to result in new genotypes with different nutritional needs. Geneticists and nutritionists will have to work closely together in the future to identify nutritional needs of these birds. 3. Collaboration between nutrition and other disciplines has been important in maintaining efficient poultry production and health and preventing or alleviating a number of metabolic disorders 4. There are many new challenges facing nutritionists to maintain health and performance in flocks under more exacting standards of bird welfare and human health. 5. A vigorous contribution from nutritional science and research will continue to be important for the future well-being of the poultry industry. PMID- 10821517 TI - Effect of foraging material and food form on feather pecking in laying hens. AB - 1. The aim was to test whether provision of foraging material and food form influence feather pecking and feather damage in laying hens. 2. From week 19 of age, 16 groups of 11 hens (white Lohman Selected Leghorn hybrids) were kept in pens with or without access to long-cut straw as foraging material and fed on either mash or pellets. 3. Foraging behaviour was increased in hens with access to straw and time spent feeding was increased in hens fed on mash. In addition, hens fed on mash had longer feeding bouts and higher rates of pecking at the food during feeding than hens fed on pellets. 4. There were interaction effects of foraging material and food form on both feather pecking and feather damage. High rates of feather pecking and pronounced feather damage were only found in hens housed without access to straw and fed on pellets. In groups characterised by high rates of feather pecking the hens also showed more severe forms of this behaviour. 5. Differences in the time budgets of hens kept in different housing conditions suggested that birds fed on mash used the food not only for feeding but also as a substrate for foraging behaviour. 6. In order to avoid problems with feather pecking it is recommended that laying hens are provided with foraging material and fed on mash. PMID- 10821518 TI - Stress and feather pecking in laying hens in relation to housing conditions. AB - 1. Possible association between high rates of feather pecking and increased stress were investigated in laying hens. 2. From week 19 to week 30 after hatching, 16 groups of 11 hens (white Lohman Selected Leghorn hybrids) were kept in pens with or without long-cut straw as foraging material and provided with food in the form of pellets or mash. 3. Stress was assessed by egg production, weight gain, tonic immobility (TI), heterophil/lymphocyte (H/L) ratio and antibody titres to sheep red blood cells (SRBC), tetanus toxoid (TT) and human serum albumin (HSA). 4. Provision of foraging material and food form influenced feather pecking. Rates of feather pecking were highest in groups housed without straw and fed on pellets. 5. Egg production was reduced in pens without straw but not affected by food form. Both the duration of TI and H/L ratios were influenced by provision of foraging material and food form. TI was longer and H/L ratios were increased in hens housed without straw and in those fed on pellets. Antibody titers to SRBC and TT were lower in pens without straw than with straw but not influenced by food form. 6. In conclusion, foraging material and food form affected both feather pecking and indicators of stress, suggesting that feather pecking in laying hens is associated with stress. PMID- 10821519 TI - Measurement of within and between population genetic variability in quails. AB - 1. To detect polymorphism in various quail lines, Randomly Amplified Polymorphic DNA (RAPD) markers were tested and were found to be effective. 2. Twenty decamer primers were selected at random and tested, and 6 of these generated distinct polymorphic patterns between the quail lines. 3. Out of a total of 60 bands amplified using 6 selected primers, 19 (31.7%) were found to be polymorphic. 4. Genetic similarity within the lines ranged from 0.726 in HBW to 0.836 in KLQ. 5. Genetic similarity estimates between the populations ranged from 0.709 to 0.808. PMID- 10821520 TI - Inheritance of bone characteristics affecting osteoporosis in laying hens. AB - 1. Heritabilities of a range of morphometric, radiological and strength characteristics were measured in the bones of end-of-lay hens. 2. Tibial strength (TSTR), humeral strength (HSTR) and keel radiographic density (KRD) were moderately to strongly inherited and were combined in a Bone Index which was used as a basis for selection. Data are available on 6 generations/cohorts of hens (n=1306), the last 3 of which are the progeny of divergently selected birds. 3. All bone characteristics used in the Bone Index responded rapidly to divergent selection and were strongly correlated with each other. In the last generation, the lines differed by 25% for TSTR, 13% for HSTR and 19% for KRD. The heritability of the index was 0.40. 4. There were no apparent genotype by environment interactions between birds housed at 2 different locations. 5. The incidence of bone fractures was significantly decreased in the line selected for high bone strength compared to the line selected for low bone strength. Humerus fracture incidence differed by a factor of 6 between the lines in the last generation. There was a strong quadratic relationship between tibia strength and overall fracture incidence (r2=0.92, P<0.01). 6. The results imply that selection for enhanced bone strength can be used as a long-term strategy for alleviating the problems of osteoporosis in laying hens. PMID- 10821521 TI - Carcase characteristics of a heavy Japanese quail line under introgression with the roux gene. AB - 1. The roux plumage sex-linked recessive gene may be used for early sexing of Japanese quail in crossbreeding production systems with wild-type and roux lines. However, associated effects of the gene on carcase and meat composition need to be assessed. 2. Quail carcases from pure Line K males and females (100% K), a heavy meat line which was used as the recipient line for the introgression of the roux gene, and from same-age roux or wild-type females from the second generation of introgression (75% K) were dissected. The effects of sex, line and plumage colour on carcase components and on protein and lipid contents of various tissues were estimated. 3. Expected sex differences in carcase weights were obtained, with marginally higher values for females. However, weights of parts and carcase yield (ratio of empty carcase weight without head, neck and feet over live body weight) were similar for both sexes in pure Line K which had a 68% carcase yield. Lipid contents in leg meat were higher in males (3.1%) than in females (2.7%). 4. The roux gene did not seem to have any major impact on carcase parts or composition. However, in roux birds, leg skin was marginally higher in lipids and pectoralis major lower in proteins than in wild-type ones. PMID- 10821522 TI - Hatching success of ostrich eggs in relation to setting, turning and angle of rotation. AB - 1. Three trials were designed to study the effects of axis of setting, turning frequency and axis and angle of rotation on the hatching success of ostrich eggs. The joint effects of axis of setting and angle of rotation were investigated in a fourth trial. 2. The hatchability of fertile ostrich eggs artificially incubated in electronic incubators (turned through 60 degrees hourly) was improved substantially in eggs set in horizontal positions for 2 or 3 weeks and vertically for the rest of the time. 3. The hatchability of fertile eggs set in the horizontal position without any turning was very low (27%). It was improved to approximately 60% by manual turning through 180 degrees around the short axis and through 60 degrees around the long axis at 08.00 and 16.00 h. A further improvement to approximately 80% was obtained in eggs automatically turned through 60 degrees around the long axis in the incubator. Additional turning through 180 degrees around the short axis twice daily at 08.00 and 16.00 h resulted in no further improvement. 4. The hatchability of fertile eggs set vertically in electronic incubators and rotated hourly through angles ranging from 60 degrees to 90 degrees around the short axis increased linearly over the range studied. The response amounted to 1.83% for an increase of 10 (R2=0.96). 5. The detrimental effect of rotation through the smaller angle of 60 degrees around the short axis could be compensated for by setting ostrich eggs in the horizontal position for 2 weeks before putting them in the vertical position. PMID- 10821523 TI - Relationship between early post-mortem muscle pH and shortening-induced toughness in the Pectoralis major muscle of processed broilers air-chilled at 0 degrees C and -12 degrees C. AB - 1. An experiment was conducted to investigate the development of shortening induced toughness in the Pectoralis major (PM) muscles of commercially processed broilers, air-chilled at 0 degrees C and -12 degrees C, as a function of muscle pH early post-mortem. Electrical stimulation was used immediately after stunning and neck cutting to provide carcases with pH values 15 min post-mortem (pH15 min) ranging between 6.79 and 5.85. 2. The deep PM muscle temperatures of carcases chilled at -12 degrees C were lower (cooler) after primary chilling and at 215 min post-mortem than those chilled at 0 degrees C, although chilling regimen had no major effect on pH values over the 24 h post-mortem period. However, carcases chilled at -12 degrees C had longer sarcomeres, lower cooking losses and lower shear force values than those chilled at 0 degrees C. 3. Correlation analysis of the results for both chilling regimens clearly demonstrated that over the pH15min range 6.79 to 5.85, carcases with the lowest pH15min values had the shortest sarcomeres, the highest cooking losses and the toughest meat. In addition, there was no evidence to support the occurrence of cold shortening within this population. This suggests that an early onset of rigor at higher temperatures in broiler carcases, as well as inducing rigor shortening and toughness, might also induce greater protein denaturation and subsequent loss of water holding capacity as manifested in increased cooking losses. 4. Quadratic regression curves showed that over the pH15min range 6.80 to 6.30, only the fast chilling regimen at -12 degrees C could inhibit rigor shortening and minimise changes in cooking loss and shear force values. However, neither chilling regimen was effective in preventing severe rigor shortening, increased cooking losses and adverse toughness in carcases with pH15min values below 6.30. 5. The benefits of fast chilling carcases with pH15min values above 6.3 can also be quantified in terms of carcases exceeding a 4.00 kg/cm2 toughness threshold. Only 1.9% of these carcases chilled at -12 degrees C exceeded this limit (maximum shear force value of 4.72 kg/cm2) compared to 34.9% of the carcases chilled at 0 degrees C (maximum shear force value of 8.46 kg/cm2), further emphasising the considerable reduction in textural variability and improvement in tenderness gained by fast air-chilling at -12 degrees C. PMID- 10821524 TI - The metabolic use of energy from dietary fat in broilers is affected by fatty acid saturation. AB - 1. Two experiments were performed to evaluate the effect of dietary fat on performance and fat and protein accretion in broiler chickens according to the degree of saturation. 2. The first experiment was designed to test 2 sources of dietary fat and 3 levels of dietary energy using a factorial (2x3) experimental design. The foods were formulated to maintain a constant ratio of energy to protein (and other nutrients). There were no significant differences in weight gain, intake, final body weight or food to gain ratio between broilers fed on diets differing solely in the degree of fat saturation. Broilers fed on diets containing animal fat showed higher whole-body fat retention (P=0.02) and lower protein accretion (P=0.03) than those fed on diets containing vegetable oils. 3. In the second experiment, only 1 concentration of fat (tallow, lard or sunflower oil) was incorporated into the experimental diets, providing different energy to protein ratios. The carcase protein content was not affected by dietary fat source, while total fat accretion (P=0.01) and energy retention (P=0.14) were highest in broilers fed on a diet containing tallow. 4. The findings suggest that the degree of saturation of dietary fats affects their metabolic use and fat accumulation in broiler chickens. PMID- 10821525 TI - Effect of removal of caecal contents on nitrogen utilisation and nitrogen excretion in caecally ligated chickens fed on a low protein diet supplemented with urea. AB - 1. The effect of washing out the caecal contents on nitrogen utilisation and nitrogen excretion were examined in Single Comb White Leghorn cockerels fed on a 50 g/kg protein diet supplemented with urea. 2. Flushing out the caecal contents with saline in caecally ligated chickens produced a significantly increased nitrogen balance and increased nitrogen utilisation (P<0.05). 3. Washing out the caecal contents significantly decreased uric acid excretion but the treatment had no effect on urea and ammonia excretion. 4. Caecal bacterial contents were significantly decreased by caecal ligation and decreased further by washing out the caecal contents. 5. It is concluded that nitrogen metabolism in chickens is affected by possible changes in caecal fermentation produced by preventing substances from urine and digesta from entering the caeca. PMID- 10821526 TI - Effect of caecectomy on growth, moisture in excreta, gastrointestinal passage time and uric acid excretion in growing chicks. AB - 1. The effect of caecectomy on nitrogen utilisation and excretion was examined in growing chicks fed on a commercial diet. 2. Caecectomy had no significant effect on food intake or body weight gain. 3. Caecectomy caused significantly higher moisture content in excreta (P<0.01). 4. Gastrointestinal passage time of digesta was significantly shorter in caecectomised chicks than in control chicks (P<0.05). 5. Caecectomy tended to improve nitrogen utilisation rate in growing chicks. 6. The treatment significantly decreased uric acid excretion (P<0.01) and excretory uric acid-N/total nitrogen excretion (P<0.01). 7. It is concluded that the effects of caecectomy on nitrogen metabolism in growing chicks are similar to those in adult chickens. PMID- 10821527 TI - Effect of food enzymes on utilisation of lupin carbohydrates by broilers. AB - 1. The effects of 3 commercial enzyme products on the nutritive value of 2 lupin species were investigated with the emphasis on changes in composition of non starch polysaccharides (NSPs) along the digestive tract. Enzyme A contained primarily cellulase, beta-glucanase and xylanase activities, enzyme B primarily hemicellulase, pentosanase and xylanase activities, and enzyme C primarily hemicellulase, pectinase and beta-glucanase activities. 2. The enzymes were added to semi-purified diets based on sorghum and casein containing 35% whole seed lupins (Lupinus angustifolius cv Gungurru or Lupinus albus cv Kiev mutant). Control diets contained no lupins. 3. Food conversion ratio (FCR), excreta moisture content and apparent metabolisable energy (AME) were affected by lupin species but not by enzyme supplementation. 4. In diets with L. angustifolius, enzyme C significantly increased digesta viscosity and increased the concentration of soluble NSPs in all sections of the intestine. 5. Digestibility of protein and NSPs in the ileum and microbial fermentation in the ileum and caeca were not affected by adding enzymes to diets containing L. angustifolius. 6. Enzyme addition to diets with L. albus did not affect digesta viscosity nor concentration of soluble NSPs but caused a significantly (P<0.05) reduced concentration of insoluble NSP in the ileum. 7. Enzyme addition to L. albus significantly (P<0.05) increased NSP digestibility in the ileum but had no effects on protein digestibility and fermentation in the ileum and caeca. PMID- 10821528 TI - Effect of supplemental methionine, choline and their combinations on the performance and immune response of broilers. AB - 1. A study was conducted with broilers to find out the effect of addition of methionine (0.0, 1.5, 3.0 and 4.5 g/kg) and choline (0.0, 1000 and 2000 mg/kg) to a basal diet containing crude protein, 221 g/kg, ME, 12.25 MJ/kg, methionine, 3.6 g/kg and choline, 1300 mg/kg. 2. Supplemental levels of methionine and choline were found to be ineffective in improving the growth, food consumption and food conversion efficiency of broilers. 3. However, a higher value for leucocyte migration inhibition was observed in chicks fed on the diet containing 6.5 g/kg methionine and 1300 mg/kg choline, indicating a significantly improved cellular immune response. 4. HI test and ELISA indicated enhanced antibody titres in chicks receiving 3.0 g/kg methionine and 3300 mg/kg choline, showing a significantly better humoral immune response. 5. Methionine and choline should be supplemented at levels higher than the recommended level of BIS for better health and production in chicks. PMID- 10821529 TI - Embryonic heart rate measurements during artificial incubation of emu eggs. AB - 1. Daily changes in embryonic heart rate (HR) of emu were determined non invasively at 36 degrees C by acoustocardiography (ACG) during the last 30% of artificial incubation (predicted incubation time is 50 d). 2. The pattern of daily changes in mean HR of hatched embryos decreased from about 175 bpm to about 140 bpm towards the end of incubation. 3. The mean HR at 80% of incubation (ca. 170 bpm) was close to the value predicted from an allometric equation reported previously for precocial domesticated birds. 4. ACG could measure embryonic HR even during the external pipping period. 5. If the artificial external pipping procedure is timed correctly after internal pipping, it might aid the embryos in hatching. However, further investigation into this aspect is needed. PMID- 10821530 TI - Relative humidity at moderate ambient temperatures: its effect on male broiler chickens and turkeys. AB - 1. The effect of relative humidity (RH, 40% to 75%) at moderate ambient temperatures (Ta, 28 degrees and 30 degrees C) on the performance and thermoregulation of male broiler chickens and turkeys was studied at the age of 4 to 8 weeks. 2. Weight gain and food intake of male broiler chickens were significantly higher at 60% to 65% RH but food conversion efficiency was not affected by RH. In male turkeys, no effect of RH on performance was recorded. 3. Both chickens and turkeys controlled body temperature at normothermic levels during exposure to the experimental environmental conditions. 4. The rate of panting estimated from blood pH and pCO2 was lower in chickens exposed to 28 degrees C than to 30 degrees C. In turkeys, the rate was lower than that recorded in chickens at both Ta(s). 5. Plasma T3 was positively and significantly correlated with food intake. 6. It may be concluded that RH plays a role in the performance of chickens exposed to 30 degrees and 28 degrees C. whereas male turkeys must respond to RH at Ta >30 degrees C. PMID- 10821531 TI - The effect of light intensity on growth and development of turkey toms. AB - 1. The effect of light intensities from 10 to 700 lux on the performance of 5 to 18 week-old turkey males was studied in 2 trials. 2. Body weight of 18 week-old turkeys, in both experiments, was highest under the lowest light intensity This coincided with higher weight gain and lower food intake, which resulted in significantly better food conversion efficiency 3. Light intensity affected heart muscle weight but not weight of breast muscle, abdominal fat or testis as proportions of body weight. 4. The decline in plasma T3 concentration with age differed from other treatments at the low light intensity, which resulted in a significantly higher T3 concentration in turkeys exposed to 10 lux at the age of 10 to 15 weeks. 5. It is concluded that light intensity significantly affects food conversion efficiency in turkey males. This is likely to be related to differential investment of energy expenditure for maintenance. PMID- 10821532 TI - Food deprivation and feeding of broiler chickens is associated with rapid and interdependent changes in the somatotrophic and thyrotrophic axes. AB - 1. In several experiments, hormonal changes in the somatotrophic axis, growth hormone (GH) sensitivity to a GH-secretagogue, thyroid hormones and their metabolising enzymes and plasma glucose levels were measured in relation to food deprivation and reinitiation after a single daily meal in 4- to 5-week-old male broiler chickens. 2. Floor-reared male broiler chickens were fed ad libitum or were restricted to a daily food intake of 40 or 45 g per d from the age of 2 weeks onwards. The daily food allowance was consumed in 0.5 h. 3. Food deprivation increased plasma GH concentrations but decreased GH-dependent variables such as plasma insulin-like growth factor-I and 3,3',5-triiodothyronine (T3) concentrations. Hepatic inner ring deiodinating type III activity was markedly elevated, presumably as a consequence of low hepatic GH receptor numbers, and is thought to be the causal mechanism for the low plasma T3 concentrations. Food intake reversed these variables in a time-related manner. 4. GH pulsatility characteristics, as calculated by deconvolution analysis, revealed profound changes between food restricted and ad libitum fed animals. Chickens deprived of food for about 23.5 h were characterised by an enhanced pulsatile GH release as reflected in the higher GH secretory burst amplitude, GH mass per burst, GH production rate and GH pulse frequency. These variables returned very quickly to normal values after refeeding. 5. In summary these experiments taken together demonstrate very clearly the interdependent and time-related changes of the somatotrophic and thyroid axes upon a single meal in previously food-deprived broiler chickens. PMID- 10821533 TI - The role of mismatch repair in DNA damage-induced apoptosis. AB - DNA mismatch repair plays a critical role in maintaining genomic integrity. Defects in human mismatch repair are the primary cause of certain types of cancer, including hereditary nonpolyposis colorectal cancer. In the past, the ability of mismatch repair proteins to correct DNA mismatches that occur during DNA replication, repair, and recombination was considered the primary mechanism by which it contributes to genomic stability. However, increasing evidence supports the idea that the mismatch repair system also contributes to genome stability by stimulating DNA damage-induced apoptosis as part of the cytotoxic response to physical and chemical agents. MutS/MutL homologues mediate the process of apoptosis by binding to DNA adducts and either provoking futile repair events or blocking steps in DNA metabolism (i.e., DNA replication and/or repair). This damage recognition step by mismatch repair (MMR) proteins stimulates a signaling cascade for apoptosis, resulting in activation of protein kinase(s) that phosphorylate p53 and/or the related protein p73. Activated p53 and p73 in turn transmit a signal to the apoptotic machinery to execute cell death. The goal of this commentary is to discuss the molecular mechanism(s) by which mismatch repair proteins stimulate apoptosis. PMID- 10821534 TI - Targeted antiestrogens for the prevention of breast cancer. AB - This commentary explores the recent experience with and the basis for the use of selective estrogen receptor modulators (SERMs) to prevent breast cancer. Chemoprevention has been a goal for many years. As newer agents are unveiled, they will continue to be tested against tamoxifen, the current standard for the treatment and prevention of breast cancer. Raloxifene holds the promise of treating osteoporosis with the beneficial side effect of breast cancer prevention. The Study of Tamoxifen and Raloxifene (STAR) trial and prior prevention studies will be discussed in an attempt understand the bridge from the laboratory to the clinic. PMID- 10821535 TI - Expression of ribosomal protein S5 cloned gene during differentiation and apoptosis in murine erythroleukemia (MEL) cells. AB - Murine erythroleukemia (MEL) cells have been used as a suitable model system for studying cellular and molecular mechanisms of erythroid differentiation. In an effort to isolate and characterize genes whose expression change during differentiation, we cloned and sequenced a cDNA of 715 bp (rpS5) from a MEL cDNA library. The cloned mouse cDNA showed significant degree of structural homology in both DNA and protein level to known human and rat genes that encode the S5 proteins of 40S ribosomal subunit. The use of 715-bp cDNA as probe revealed the presence of an RNA transcript in the cytoplasm of MEL and human neuroectodermal RD/TE-671 cells. The steady-state accumulation level of this RNA transcript decreased upon induction of differentiation of both cell lines by treatment with DMSO and UDP-4, two structurally different inducers. Blockade of MEL cell differentiation by the inhibitor N6-methyladenosine preserved the constitutive expression of the rpS5 gene. DNA methylation analysis at CCGG sites located at the rpS5 gene locus in undifferentiated and differentiated MEL cells revealed that the suppression of the rpS5 gene during MEL cell differentiation is not related to any change in methylation at these sites. Moreover, the rpS5 gene continued to be expressed in cells undergoing serum-deprived apoptosis, like in control untreated cells. Therefore, we conclude that there may be a disparate pattern of expression of the rpS5 gene in differentiating and apoptotic cells. These data can be valuable in understanding the role of ribosomal proteins during differentiation and cell death (apoptosis) of neoplastic cells, although there is no experimental evidence that the suppression of the rpS5 gene is related mechanistically to the induction of differentiation. It may well be considered as part of the differentiation process. PMID- 10821536 TI - Human N-ras, TRK-T1, and RET/PTC3 oncogenes, driven by a thyroglobulin promoter, differently affect the expression of differentiation markers and the proliferation of thyroid epithelial cells. AB - The ras family members and the tyrosine kinases RET and TRK are frequently activated in human tumors of the thyroid gland. To ascertain the effects of these oncogenes in cultured thyroid cells we have generated expression vectors containing activated versions of the three genes under the control of the thyroid specific thyroglobulin gene promoter. Here we show that the expression of the three oncogenes differently affects thyroid differentiation. While the TRK-T1 oncogene interferes with the capability of thyroid cells of trapping iodide and only marginally affects thyroglobulin gene expression, both RET/PTC3 and N ras(Gln61-Lys) induce a dramatic reduction of thyroglobulin mRNA and alleviate TSH dependency for cellular growth. However, none of the three oncogenes is able to induce the appearance of neoplastic transformation markers, such as growth in semisolid medium and tumorigenicity in athymic mice. This indicates that genetic events additional to TRK, RET, or N-ras activation are required for fully malignant transformation of thyroid cells. PMID- 10821537 TI - Attenuation of the induced differentiation of HL-60 leukemia cells by mitochondrial chaperone HSP70. AB - The HSP70 family of heat shock proteins, which are involved in development and cellular differentiation, is elevated in various tumor cell lines. To examine the role of these proteins in neoplastic cell differentiation, four members of the HSP70 multiple gene family (i.e., HSP70, HSC70, GRP78, and mtHSP70) were examined during the induced differentiation of HL-60 promyelocytic leukemia cells. Western analyses showed that continuous exposure for 48 h of HL-60 cells to the differentiation-inducing agents, all-trans retinoic acid, 1,25-dihydroxyvitamin D3, or N-methylformamide, resulted in decreases in mitochondrial HSP70 (mtHSP70), with little change in the levels of HSP70, HSC70, and GRP78. To gain information on the role of mtHSP70 in the differentiation process, HL-60 cells were transfected with either murine mthsp70 cDNA or vector alone. Slightly greater than twofold increases in mtHSP70 protein levels occurred in cells transfected with the mthsp70 cDNA. In vector-transfected HL-60 cells, myeloid differentiation, measured as an increase in CD31 expression and nitroblue tetrazolium positivity, was observed following 3-6 days of treatment with each of the three inducing agents. In contrast, cell differentiation induced by each agent was markedly attenuated in mthsp70-transfected HL-60 cells. These findings suggest that a decrease in mtHSP70 is important for the induced differentiation of HL-60 cells. PMID- 10821538 TI - Marked variation of thymidylate synthase and folylpolyglutamate synthetase gene expression in human colorectal tumors. AB - Patients with advanced colorectal cancer are currently being treated with 5 fluorouracil (5-FU)-based chemotherapy. A growing number of patients with resectable disease receive adjuvant therapy with 5-FU/levamisole (LEV) or 5 FU/folinic acid (LV). However, many patients still fail on these treatments, due to occurrence of natural or acquired tumor resistance. Among clinically relevant mechanisms of resistance to fluoropyrimidines, increased expression of thymidylate synthase (TS) has been emphasized. Another potentially relevant mechanism involves a decrease in folylpolyglutamate synthetase (FPGS) expression. To establish the value of these genes as prognostic factors and predictors of the outcome of 5-FU-based chemotherapy in colorectal cancer, we measured their expression in colorectal tumors from patients undergoing surgery and postoperative chemotherapy and compared it with that in normal colonic mucosa. This was done by a semi quantitative, nonradioisotopic polymerase chain reaction (PCR) method using beta-actin as an internal standard and expressed as a TS/beta actin or a FPGS/beta-actin mRNA ratio. In tumor samples from 21 colorectal cancer patients, TS gene expression varied 118-fold. The median TS/beta-actin ratio was, in fact, 41.36 x 10(-3) (range 2.49 x 10(-3) to 294.54 x 10(-3)). Little variation in TS gene expression was observed in corresponding normal colic mucosa; the TS/beta-actin gene ratio was lower (median 26.16 x 10(-3); range 8.49 x 10(-3) to 69.49 x 10(-3)). Among tumor explants from 20 patients, FPGS expression varied over 161-fold. A similar marked variation was also observed in normal colonic mucosal samples (over 185-fold). Overall and disease-free survival data suggest an inverse association between the level of tumor TS and FPGS expression and clinical prognosis. The availability of this sensitive and accurate assay for gene expression should now make it possible to extend these laboratory/clinical correlations to larger populations. PMID- 10821539 TI - Ossicular chain reconstruction using a new tissue adhesive. AB - HYPOTHESIS: A new medical-grade cyanoacrylate tissue adhesive will improve the results of ossicular chain reconstruction in a rat model. BACKGROUND: An ideal tissue adhesive has long been awaited by otologists. Studies examining the older cyanoacrylates have demonstrated variable efficacy and toxicity. Octylcyanoacrylate is a new tissue adhesive that has many ideal properties for otologic surgery. METHODS: Thirteen female C-D rats were anesthetized, and preoperative auditory brainstem response (ABR) testing was performed. A left antrotomy was performed, and the incus was removed. In the adhesive group, the incus was dipped in octylcyanoacrylate and interposed between the tympanic membrane and the stapes; no adhesive was used in the control group. At 8 weeks, postoperative ABR was performed, the integrity of the ossicular chain inspected, and histopathologic analysis of the temporal bones performed. Statistical comparison of ABR results was performed with the Mann-Whitney test. RESULTS: Seven rats were randomized to the adhesive group and six to the control group, of which four survived. There were no histopathologic differences in the temporal bones of the animals other than the presence of mild foreign body reaction around the ossicular chain of the animals in the adhesive group. The ossicular chain was not intact in two of the four controls, whereas the rest were intact at 8 weeks. Postoperative air conduction ABR results (mean dB sound pressure level) (62.5 control versus 34.3 adhesive, p = 0.010) and air-bone gaps (47.5 control versus 18.9 adhesive, p = 0.008) were significantly better in the adhesive group. CONCLUSIONS: This new medical-grade tissue adhesive improves the hearing results of ossicular chain reconstruction, with no apparent histotoxicity in this animal model. PMID- 10821540 TI - Cavum major tympano-ossiculoplasty. AB - OBJECTIVE: To evaluate the success of tympano-ossiculoplasty in patients with previous canal wall down mastoidectomy. STUDY DESIGN: A retrospective review of 79 patients who underwent cavum major tympano-ossiculoplasty from a total of 1,910 tympanomastoidectomies from 1976 to 1998. OUTCOME MEASURES: The results of air-bone gap closure, surgical findings, and revision surgery are presented. RESULTS: In 63% of patients, the air-bone gap closed to within 20 dB with a mean gain of 14.7 dB. CONCLUSION: A significant percentage of patients will gain substantial improvement in their hearing after cavum major tympano-ossiculoplasty with minimal risk. PMID- 10821541 TI - Octylcyanoacrylate: a new medical-grade adhesive for otologic surgery. AB - HYPOTHESIS: The adhesive octylcyanoacrylate is not associated with significant inner ear toxicity in a guinea pig model. BACKGROUND: Many cyanoacrylate adhesives have been investigated for use in otologic surgery, but variable ototoxicity has been reported. Octylcyanoacrylate is a medical-grade adhesive with many properties that make it ideal for use in the ear. It is free of contaminants; it forms a strong, flexible bond; and it inhibits the growth of gram-positive organisms in culture. This is the first study to assess the ototoxicity of this new adhesive. METHODS: Fourteen adult guinea pigs were used. Preoperative auditory brainstem responses (ABRs) were determined. Bilateral antrotomies were performed, and the ears were randomized to adhesive and control (saline) groups. In the adhesive ears. 0.5 or 0.1 mL of octylcyanoacrylate was instilled into the middle ear. Eight weeks later, postoperative ABRs were determined, the animals were killed, and the temporal bones were removed. Middle ear changes were noted, and the ossicular chain was assessed. Cochlear hair cell analyses were performed. Histologic assessment of the middle ear mucosa was performed. RESULTS: There was a higher incidence of conductive hearing loss in the adhesive group secondary to fixation of the ossicular chain, but there was no significant difference in bone conduction thresholds. The median postoperative bone conduction thresholds (dB peak sound pressure level) was 15.0 in the control group and 17.5 in the adhesive group, p = 0.89. There was also no significant difference in inner hair cell counts (0.4% vs. 0.5% median hair cell loss, p = 0.72) or outer hair cell counts (3.7% vs. 3.0% median hair cell loss, p = 0.23) for the adhesive and control groups, respectively. Histopathologic analysis of the middle ear mucosa demonstrated variable mild to moderate foreign body reaction with no evidence of mucosal ulceration or necrosis. CONCLUSIONS: A large amount of octylcyanoacrylate placed in the middle ear of the guinea pig did not cause any morphologic or functional evidence of inner ear toxicity. This new adhesive is a promising tool for otologic surgery. PMID- 10821542 TI - Surgical management of the atelectatic ear. AB - OBJECTIVE: The surgical management of the atelectatic ear is controversial because the natural course toward cholesteatoma development cannot be predicted, and hearing acuity remains normal until later in the disease course. Consequently, surgery is often delayed until there is a clear indication, such as hearing loss or frank cholesteatoma development, but such delay often necessitates more extensive surgery. Because earlier intervention appears to be in the best interest of the patient but is often avoided because of near normal hearing levels at this stage, the author proposes a staging system for classification and management of the atelectatic ear. Hearing results and complications in patients undergoing tympanoplasty with or without ossicular reconstruction are reported for patients with type III and IV retractions. STUDY DESIGN: A retrospective study using a computerized otologic database to identify patients who meet the inclusion criteria. SETTING: A tertiary referral center. PATIENTS: A total of 55 patients (63 ears) aged 5 to 78 years underwent cartilage tympanoplasty with or without ossicular reconstruction. INTERVENTIONS: Elevation of the ear drum, followed by cartilage reconstruction of the tympanic membrane, with ossicular reconstruction as indicated. MAIN OUTCOME MEASURES: Postoperative pure tone average air-bone gap for four frequencies (500, 1000, 2000, 4000 Hz) compared with preoperative levels. RESULTS: There was a statistically significant improvement in hearing (p < 0.05). CONCLUSIONS: This staging system offers an effective treatment algorithm for pars tensa retractions and management of type III and IV retractions via cartilage tympanoplasty with or without ossicular reconstruction and is a proven treatment modality. PMID- 10821543 TI - Experimental investigations of the use of cartilage in tympanic membrane reconstruction. AB - BACKGROUND: Temporalis fascia, perichondrium, and cartilage are commonly used for reconstruction of the tympanic membrane in middle ear surgery. Cartilage grafts offer the advantage of higher mechanical stability, particularly in cases of chronic tubal dysfunction, adhesive processes, or total defects of the tympanic membrane, in contrast to fascia and perichondrium, which presumably offer better acoustic quality. HYPOTHESIS: The purpose of this study was to determine the acoustic transfer characteristics of cartilage of varying thickness and its mechanical deformation when exposed to fluctuations in atmospheric pressure. METHOD: Ten pairs of cartilage specimens from the cavum conchae and the tragus were obtained from fresh human cadavers. Young's modulus was determined by mechanical tension tests and statistically evaluated using the t test. The acoustic transfer characteristics of an additional 10 specimens were measured by a laser Doppler Interferometer after stimulation with white noise in an external auditory canal--tympanic membrane model. Mechanical stability was determined by measuring displacement of the cartilage using static pressure loads of < or = 4 kPa. RESULTS: Young's modulus determinations for conchal and tragal cartilage were 3.4 N/mm2 and 2.8 N/mm2, respectively, but the difference was not significant. Acoustic testing showed a 5-dB higher vibration amplitude in the midfrequency range for conchal compared with tragal cartilage, but the difference was not significant. Reducing cartilage thickness led to an improvement of its acoustic transfer qualities, with a thickness < or = 500 microm resulting in an acceptable acoustic transfer loss compared with the tympanic membrane. CONCLUSION: Both conchal and tragal cartilage are useful for reconstruction of the tympanic membrane from the perspective of their acoustic properties. The acoustic transfer loss of cartilage can be reduced by decreasing its thickness. A thickness of 500 microm is regarded as a good compromise between sufficient mechanical stability and low acoustic transfer loss. PMID- 10821544 TI - Performance of multiply handicapped children using cochlear implants. AB - OBJECTIVE: To explore the possibility of expanding implantation guidelines to include the multiply handicapped population and to determine the pattern of development of perceptual skills in deaf children who have other impairments. PATIENTS AND SETTING: The study group consisted of 31 profoundly deaf children with documented impairments in addition to deafness who were consecutively implanted. Twenty-nine of the children underwent implantation and were followed at NYU Medical Center for at least 1 year. INTERVENTIONS AND MAIN OUTCOME MEASURES: Routine audiometric measurements and closed- and open-set measures of speech perception were performed at the phoneme, word, and sentence level pre- and postoperatively. Analyses of variance were performed when possible. RESULTS: The study population showed substantial improvement after implantation in the ability to perceive phonemes, words, and sentences using audition alone. CONCLUSIONS: Multiply handicapped children obtain demonstrable benefit from cochlear implantation, with no increase in surgical complications, although the rate of growth of perceptual skills is slower than for deaf child with no additional complicating factors. PMID- 10821545 TI - Cochlear implantation after labyrinthectomy. AB - OBJECTIVE: The goal was to report a method used for intraoperative ear selection for cochlear implantation using electrical brainstem response. Initial patient response and the longer-term results of cochlear implantation after labyrinthectomy were compared. STUDY DESIGN: This was a specific retrospective review of a single case of cochlear implantation after labyrinthectomy. SETTING: The study involved a tertiary referral center in both an ambulatory and a hospital setting. PATIENT: The study involved a report of a single patient who was evaluated for a possible cochlear implant and successfully underwent cochlear implantation. INTERVENTIONS: A case study of a profoundly deaf individual is presented, including the diagnostic measures used to determine the candidacy for cochlear implantation, the ear selected, and the rehabilitation. MAIN OUTCOME MEASURES: Both early (3 months) and later (14 months) postoperative results clearly demonstrate that a cochlear implant in a patient with a previous labyrinthectomy can be beneficial. RESULTS: The early and later results after cochlear implantation are compared in a single case study. CONCLUSION: This case study demonstrates that there is improvement in sound awareness, speech recognition, and communication after cochlear implantation in a previously labyrinthectomized ear. PMID- 10821546 TI - Child and family factors associated with deaf children's success in auditory verbal therapy. AB - OBJECTIVES: To identify the general demographics of children who had Auditory Verbal therapy and to identify child and family factors associated with differences between those children for whom Auditory-Verbal therapy led to success and those for whom it did not. SETTING: Private tertiary care facility. POPULATION: Children who had hearing losses ranging from mild to profound. INTERVENTION: Auditory-Verbal therapy, a therapeutic intervention designed to teach parents to educate their young deaf and hearing-impaired children to use residual hearing and to speak, was used. MAIN OUTCOME MEASURES: Clinic files, parent questionnaires, and parent report of current success were used to determine efficacy of treatment. RESULTS: Fifty-seven percent of the clients who remained in this program for over 1 year were fully integrated into regular education, with no services from a teacher of the deaf. The population was affluent, with more females than expected. Those who left dissatisfied tended to be males with greater degrees of hearing loss who left the program soon after 1 year. CONCLUSIONS: Auditory-Verbal therapy provides successful intervention to students with a particular set of demographic characteristics. PMID- 10821547 TI - Cochlear deafness in a Chinese family with Fechtner's syndrome. AB - OBJECTIVE: To identify the nature of the hearing impairment in the members of a Chinese family with Fechtner's syndrome. STUDY DESIGN: Retrospective case review. SETTING: Tertiary referral center. PATIENTS: A Chinese family with a variant of Alport's syndrome: high-tone sensorineural hearing loss, proteinuria, macrothrombocytopenia, and ocular disease. INTERVENTIONS: The diagnosis of Fechtner's syndrome was confirmed by the characteristic ultrastructure of the Dohle-like inclusion bodies in the neutrophils of the mother and her three children. Pure-tone audiometry, evoked response audiometry (ERA), and distortion product otoacoustic emissions (DPOAE) were performed in two subjects to investigate the hearing impairment. MAIN OUTCOME MEASURE: The parameters of the ERA and DPOAEs were correlated. RESULTS: In both subjects, the ERA was within normal limits, and there were no measurable DPOAEs in frequencies >2 kHz. CONCLUSION: The hearing loss in Fechtner's syndrome is cochlear rather than neural. PMID- 10821548 TI - External aperture of the vestibular aqueduct in Meniere's disease. AB - OBJECTIVE: To study the relationship of the length of the external aperture of the vestibular aqueduct and the ratio of the summating potential and action potential (SP:AP) in patients with Meniere's disease. STUDY DESIGN: Retrospective case study. SETTING: Neurotology referral center. PATIENTS: Fifty-four patients with Meniere's disease and nine control subjects without Meniere's disease. INTERVENTION: The external aperture of the vestibular aqueduct was measured from a three-dimensional surface reconstruction computed tomography scan. Transtympanic electrocochleography was performed on patients with Meniere's disease. MAIN OUTCOME MEASURE: The length of the external aperture of the vestibular aqueduct in the Meniere's disease ears was related to the SP:AP ratio in the Meniere's disease ears and compared with controls. RESULTS: The average length of the external aperture was 3.79 +/- 2.92 mm in Meniere's disease ears and 5.35 +/- 1.73 mm in the control ears (p < 0.05). An enlarged SP:AP ratio was found in 95% of ears in the group with nonvisible external apertures of the vestibular aqueduct, 91% of ears in the <5 mm group, 58% of ears in the 5-7 mm group, and 29% of ears in the >7 mm group (chi-square = 24.814; p = 0.000). CONCLUSIONS: The length of the external aperture of the vestibular aqueduct in patients with Meniere's disease is significantly shorter than in those without Meniere's disease. Endolymphatic hydrops, evidenced by an enlarged SP:AP ratio, was related to the length of the external aperture of the vestibular aqueduct. The shorter the external aperture, the more often the SP:AP ratio was enlarged. A short or nonvisible external aperture of the vestibular aqueduct is a predisposing factor to the development of Meniere's disease. PMID- 10821549 TI - The effect of the canalith repositioning maneuver on resolving postural instability in patients with benign paroxysmal positional vertigo. AB - OBJECTIVE: Patients with benign paroxysmal positional vertigo (BPPV) often experience postural instability as well as brief episodes of vertigo. The purpose of this study was to determine whether successful resolution of the episodic vertigo, through use of the canalith repositioning treatment, would be accompanied by improvement in postural stability. STUDY DESIGN: Prospective clinical study. SETTING: Outpatient tertiary care facility in a university. PATIENTS: Thirty-three patients with a diagnosis of the canalithiasis form of BPPV affecting the posterior canal unilaterally. All patients had complete remission of the positional vertigo after treatment. Patients with abnormal caloric or rotary chair test results were excluded from the study. INTERVENTION: The posterior canal BPPV was treated by the canalith repositioning treatment. MAIN OUTCOME MEASURES: Postural stability was assessed by computerized dynamic posturography before and 1 to 2 weeks after treatment. Six different subtests were used. RESULTS: A significant number of patients had abnormal stability, as measured with computerized dynamic posturography, before treatment. After treatment there was a significant increase in the number of subjects with normal results on the different subtests; however, not all patients had normal postural stability. Younger subjects were more likely to show improved stability. CONCLUSIONS: Treatment of BPPV using the canalith repositioning treatment results in improved postural stability in patients with BPPV. Not all patients have normal stability after treatment, however, and assessment and treatment of the balance problems may be necessary. PMID- 10821550 TI - Intracranial sarcoma in a patient with neurofibromatosis type 2 treated with gamma knife radiosurgery for vestibular schwannoma. AB - OBJECTIVE: To discuss the possible relationship between stereotactic radiation therapy and the development of a meningosarcoma. STUDY DESIGN: Retrospective case review. PATIENT: A 19-year-old woman with bilateral vestibular schwannomas (neurofibromatosis type 2). One large tumor was removed totally by the translabyrinthine approach; the other smaller tumor was treated with stereotactic radiation (SRT). Six years after SRT, a malignant tumor (meningosarcoma) developed at the exact site of radiation. The patient subsequently died of this tumor. OUTCOME MEASURE: On the basis of literature surveys, the possibility and risk of postirradiation neoplasia after SRT is discussed. Furthermore, the possible causal association between SRT and the development of the meningosarcoma in this case is evaluated. CONCLUSION: On the basis of statistical considerations, the development of the reported mesenchymal sarcoma was most likely caused by the stereotactic radiation therapy. PMID- 10821551 TI - Stereotactic radiosurgery for acoustic neuromas: a survey of the American Neurotology Society. AB - OBJECTIVE: The purpose of this study was to better understand the complications, outcomes, and surgical difficulties in treating acoustic neuroma patients who have undergone stereotactic radiosurgery (SRS). STUDY DESIGN: A six-page, 28-item questionnaire was mailed to 395 members of the American Neurotology Society. SETTING: The study was conducted through an academic neurotologic practice. Questionnaire respondents were neurotologic physicians in private and academic practice. PATIENTS: A total of 46 patients who had undergone SRS were evaluated. INTERVENTIONS: Twelve (26%) of the 46 patients required microsurgery after SRS. MAIN OUTCOME MEASURES: Posttreatment cranial nerve status and the development of complications such as cerebrospinal fluid leak, meningitis, and cerebrovascular accident were evaluated. RESULTS: In the group of 12 patients who underwent microsurgery after SRS, 11 patients had some form of postoperative facial paralysis. Anacusis was present in all 12 patients. Two of the 12 patients had new-onset trigeminal neuropathy postoperatively. CONCLUSION: Microsurgical resection of acoustic neuroma after SRS is technically difficult. The difficulty exists regardless of the time of microsurgical resection after SRS. Patients who underwent microsurgery after SRS had uniformly poor cranial nerve results. PMID- 10821552 TI - Is the entire fundus of the internal auditory canal visible during the middle fossa approach for acoustic neuroma? AB - HYPOTHESIS: To determine the degree to which the fundus of the internal auditory canal (IAC) can be visualized during the middle fossa approach (MFA). BACKGROUND: Conventional wisdom states that the MFA provides excellent access to the IAC from the porus acusticus to the fundus. On the basis of observations derived from a substantial surgical experience, it became obvious that a variable fraction of the fundus lies obscure from the surgeon's line of sight during the MFA because of (1) the overhand of the transverse crest and/or (2) the immobility of the facial nerve at its entry into the fallopian canal. METHODS: Intraoperative measurements were performed in ten cases to determine the typical angle of view to the fundus of the IAC in the MFA. This angle of view was projected onto coronal computed tomography scans of 40 temporal bones. Measurements of the IAC were made to determine the amount of fundus that could not be directly visualized during a MF exposure. RESULTS: On the basis of a surgical line of sight, the fraction of the inferior compartment of the canal that could not be directly visualized because of overhand of the transverse crest ranged from 14% to 34% (median 25%). CONCLUSIONS: Complete resection of IAC tumors involving the fundus via the MFA requires some degree of blind dissection. Specialized tools and techniques are required to minimize the risk of neural injury during this indirect dissection. Inspection of the fundus with either mirror or endoscope is often necessary to exclude the possibility of retained tumor fragments. PMID- 10821553 TI - Recurrence of acoustic neuroma after incomplete resection. AB - OBJECTIVE: To determine the risk of recurrent tumor in patients after incomplete resection of acoustic neuroma. STUDY DESIGN: Retrospective case review. SETTING: A tertiary referral center. PATIENTS: Thirty-nine patients were identified who underwent incomplete resection of acoustic neuroma between January 1988 and December 1993, and had a minimum follow-up of 3.5 years, at a mean of 6.2 years (range 3.5-10.2 years). INTERVENTION: Regular follow-up. using either computed axial tomography or magnetic resonance imaging yearly. MAIN OUTCOME MEASURES: Growth of residual tumor as determined by the increase in its greatest dimension on follow-up imaging studies and the necessity for additional treatment. RESULTS: Tumor regrowth occurred in 17 patients (44%). Ten patients (26%) required additional treatment during the follow-up period. CONCLUSION: This study suggests that incomplete resection of acoustic neuroma is associated with a significant risk of recurrent tumor requiring subsequent intervention. PMID- 10821554 TI - Spontaneous involution of acoustic tumors. AB - OBJECTIVE: To determine spontaneous involution of unilateral acoustic tumors in untreated patients. STUDY DESIGN: Outcome of a continuous study, 1982 to present. SETTING: Private tertiary otology/neurotology referral center. PATIENTS: Patients with unilateral acoustic tumors for whom interval imaging was selected rather than surgery or gamma knife radiation, 1982 to the present. INTERVENTION: Interval imaging with computerized axial tomography and/or magnetic resonance imaging. MAIN OUTCOME MEASURE: Spontaneous involution of acoustic tumors. RESULTS: Forty-seven patients with unilateral acoustic tumors were untreated and followed up with interval imaging for 1 to 12.5 years. Six patients (13%), whose ages ranged from 59 to 74 years and who were followed up for 4.3 to 12.5 years, demonstrated imaging evidence of spontaneous acoustic tumor involution. Involution varied from 3.4 mm to 15 mm. CONCLUSIONS: Spontaneous involution of acoustic tumors does occur. Long-term follow-up is necessary to determine this potential. PMID- 10821555 TI - Hearing preservation in acoustic neuroma surgery: middle fossa versus retrosigmoid approach. AB - OBJECTIVE: To compare the results of the middle fossa approach with those of the retrosigmoid approach in acoustic neuroma hearing preservation surgery. STUDY DESIGN: Retrospective review. SETTING: Tertiary care facility. PATIENTS: Patients of the otology service with acoustic neuromas and useful hearing. Fifteen intracanalicular tumors were removed via a middle fossa approach and matched with 15 intracanalicular tumors removed via the retrosigmoid approach. Four additional patients with larger tumors were operated on via the middle fossa approach and matched with patients having similar tumors removed via the retrosigmoid approach. MAIN OUTCOME MEASURES: The 1994 Committee on Hearing and Equilibrium guidelines for the evaluation of hearing preservation in acoustic neuroma were applied. Facial nerve results were graded according to the House-Brackmann grading scale 3 months postoperatively. RESULTS: In the group operated on by the middle fossa approach, the average preoperative pure-tone threshold average (PTA) was 23 dB with a word recognition score (WRS) of 79%, and the postoperative PTA averaged 49 dB with a mean WRS of 56%. In the group operated on by the retrosigmoid approach, the mean preoperative PTA was 16 dB with a WRS of 95% and a postoperative PTA value of 62 dB and WRS of 51% (hearing preservation rate of 47%). The middle fossa patients had an average change in PTA of 19 dB and an average change in WRS of 20% (hearing preservation rate of 57%). Overall, the retrosigmoid patients had an average change in PTA of 42 dB and an average change in WRS of 40%. The average change in PTA for larger tumors removed via the middle fossa approach was 32 dB, whereas all matched retrosigmoid patients lost all hearing. The rate of cerebrospinal fluid leak and facial nerve outcomes were similar between the two groups. The retrosigmoid group had a higher rate of postoperative headache. CONCLUSIONS: Compared with the retrosigmoid approach, the middle fossa approach for hearing preservation surgery yields better hearing results for intracanalicular tumors and also has a lower incidence of postoperative headache. PMID- 10821556 TI - Audiometric findings in patients with acoustic neuroma. AB - OBJECTIVE: Hearing loss remains the most common symptom associated with acoustic neuroma. This study documents the audiometric findings from 721 acoustic neuroma procedures. STUDY DESIGN: This was a retrospective study. The preoperative audiometric data were compiled and were analyzed by patient age, gender, tumor size, time of surgery, and neurofibromatosis Type 2 (NF 2). Postoperative audiometric data were arranged and compiled in the same way. The hearing classification proposed by the AAO-HNS was applied to all preoperative and postoperative cases. SETTING: Tertiary referral center. PATIENTS: Surgically confirmed acoustic neuroma patients who had not previously received surgical or radiosurgical therapy. Patients underwent surgery by the retrosigmoid approach. INTERVENTION: Surgical removal of an acoustic neuroma. MAIN OUTCOME RESULT: Provision of pure tone and speech data from a group of acoustic neuroma patients, including application of the recently introduced and accepted AAO-HNS hearing classification system. RESULTS: Preoperative audiometric data were obtained from 694 of 721 patients (96%), of whom 619 had measurable hearing. Postoperative audiometry was performed on 606 patients; 152 had usable data. The combined preoperative audiometric data revealed a high frequency sensorineural hearing loss. Word recognition was servicable. The postoperative pure tones and word recognition scores were worse than preoperative scores. Age, gender, tumor size, and time of surgery had some impact on the preoperative hearing and the postoperative result; NF 2 did not. CONCLUSIONS: The study confirms that hearing alteration is almost universal in acoustic neuroma patients. Hearing preservation is possible in a significant number of cases; however, the postoperative auditory function tends to be worse. PMID- 10821557 TI - Comparison of postoperative headache after retrosigmoid approach: vestibular nerve section versus vestibular schwannoma resection. AB - OBJECTIVE: To evaluate intradural drilling as a mechanism for the development of postoperative headache after retrosigmoid craniectomy. STUDY DESIGN: A retrospective review of charts was performed on 565 retrosigmoid approaches to the cerebellopontine angle performed between January 1980 and January 1998. Patients treated with retrosigmoid vestibular nerve section without intradural drilling were compared with patients who underwent retrosigmoid removal of vestibular schwannomas in which intradural drilling was performed for exposure of the internal auditory canal. SETTING: Private practice tertiary referral center. PATIENTS: Consecutive patients undergoing retrosigmoid approach between January 1980 and January 1998 were reviewed. MAIN OUTCOME MEASURES: The presence of headache, duration of headache, and severity of headache were noted. RESULTS: In this large series, 54% of patients experienced headaches after vestibular schwannoma removal, and 5% of patients experienced headaches after vestibular nerve section (p < 0.01, chi-square). CONCLUSIONS: Postoperative headache is not a characteristic of retrosigmoid craniectomy in the absence of intradural drilling. Intradural drilling is a probable cause of headache after the retrosigmoid approach. Cranioplasty is not necessary to prevent a high incidence of postoperative headache after retrosigmoid approach. PMID- 10821558 TI - Prognostic factors for hearing preservation in vestibular schwannoma surgery. AB - OBJECTIVE: To determine whether prognostic indicators for hearing preservation could be identified in patients with vestibular schwannoma undergoing middle fossa craniotomy resection. STUDY DESIGN: Prospective case review. SETTING: Private practice tertiary referral center. PATIENTS: 333 patients with serviceable hearing and vestibular schwannoma resected by middle fossa craniotomy from 1992 to 1998. MAIN OUTCOME MEASURES: Potential prognostic indicators, including tumor size and nerve of origin, preoperative pure-tone average, speech discrimination, distortion product otoacoustic emission testing, age, auditory brainstem response (ABR), and electronystagmography. RESULTS: Postoperative hearing near preoperative levels was attained in 167 patients (50%), with an American Academy of Otolaryngology-Head and Neck Surgery Class A hearing result in 33% and a Class B result in 26%. Comparison of potential prognostic indicators between groups with hearing preserved and the group with no measurable hearing revealed significant differences in preoperative hearing, ABR, and tumor origin data. Better preoperative hearing, shorter intraaural wave V latency, shorter absolute wave V latency, and superior vestibular nerve origin were associated with higher rates of hearing preservation. CONCLUSIONS: Preoperative hearing status, ABR, and intraoperative tumor origin data were shown to be of value as prognostic indicators. PMID- 10821559 TI - Bell's palsy: a 10-year experience with antiphlogistic-rheologic infusion therapy. AB - OBJECTIVE: Treatment of idiopathic peripheral facial paralysis has remained controversial in many aspects. The authors report their experience with a protocol based on high-dose prednisolone with intravenous low-molecular-weight dextran and pentoxifylline. For this regimen, the term antiphlogistic-rheologic infusion therapy (ARIT) has been coined. STUDY DESIGN: Retrospective case-series review. SETTING: University-based hospital of otorhinolaryngology/head and neck surgery. PATIENTS: 334 patients suffering from sudden facial paralysis of unknown cause. INTERVENTION: Treatment consisted uniformly of prednisolone in a starting dosage of 250 mg tapering over 18 days and accompanying infusion of dextran and pentoxifylline. MAIN OUTCOME MEASURES: Facial nerve function after 6 months, adverse effects of therapy and comorbidity. RESULTS: From 239 patients with nonrecurrent palsy having received treatment within 12 days after onset, 92% recovered completely (House-Brackmann [HB] Grade I) without sequelae. In incomplete palsy (HB Grade II-V), normal facial function was restored in 97% of cases. Results were significantly better in the group in which therapy had been started within 3 days after the onset of palsy. Other factors such as old age, hypertension, or diabetes did not seem to influence the functional outcome in this series. Serious adverse effects requiring termination of therapy were observed in 1.2% of cases. CONCLUSION: ARIT for Bell's palsy is safe and leads to recovery rates superior to the most optimistic observations of the natural course. PMID- 10821560 TI - Imaging case of the month. Large vestibular aqueduct syndrome. PMID- 10821561 TI - Pathology case of the month. Usher's syndrome. PMID- 10821562 TI - Cracking the auditory genetic code: part II. Syndromic hereditary hearing impairment. AB - OBJECTIVE: The application of molecular genetic techniques to the study of hereditary hearing impairment has contributed significantly to our understanding of auditory physiology and disease processes. This article reviews the current state of our knowledge regarding the genes associated with syndromic hereditary hearing impairment. DATA SOURCES: Data were obtained from the Medline database and the internet. STUDY SELECTION: Articles relevant to genetics of syndromic deafness were selected. DATA EXTRACTION: Data pertaining to phenotypes, location of genes, identification of genes, and implications for hearing were extracted. CONCLUSION: Significant progress has been made in understanding the molecular pathogenesis of deafness. PMID- 10821563 TI - Intralabyrinthine schwannomas. PMID- 10821564 TI - The future of otology. PMID- 10821565 TI - Transit of radical scavenging activity of milk products prepared by Maillard reaction and Lactobacillus casei strain Shirota fermentation through the hamster intestine. AB - Oxidative stress in the colon is associated with the incidence of colon cancer. In situ, the suppression of oxidative stress in the colon would be an effective form of prevention of the cancer. In this study we investigated the transit of the radical scavenging activity of milk products through the hamster intestinal tract. Two types of skim milk products were prepared by Maillard reaction and then lactic acid fermentation. Heat treatment enhanced the radical scavenging activity for 2,2-diphenyl-1-picrylhydrazyl radical of skim milk. The activity was further increased by fermentation with Lactobacillus casei strain Shirota. Normal hamsters were fed these milk products for 14 d. For potential radical scavenging activity per unit dry weight of feces and cecal content, the groups ranked in the order of fermented product-fed hamsters > heated product-fed hamsters > control hamsters, reflecting the order of the potential of the corresponding diets. Approximately 12% of the activity of the heated and the fermented product diets passed through the gastrointestinal tract. These results suggest that some of the radical scavenging activity generated by food processing reached the colon in nonabsorbable products. PMID- 10821566 TI - Use of chemical mutagenesis for the isolation of food grade beta-galactosidase overproducing mutants of bifidobacteria, lactobacilli and Streptococcus thermophilus. AB - A classical chemical mutagenesis protocol was evaluated for increasing beta galactosidase production by probiotic bacteria to improve their potential to treat symptoms of lactose malabsorption in humans. Two Bifidobacterium species (B. breve and B. longum) and one strain each of Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus thermophilus were tested by a single exposure to two chemical mutagens, ethyl methanesulfonate (EMS) and N-methyl-N'-nitro-N nitrosoguanidine (MNNG). To screen for beta-galactosidase (beta-gal) overproducing mutants, optimized EMS and MNNG mutant pots for each strain were plated on BHI agar containing 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside (X-gal). Colonies that exhibited a blue color were selected for quantitative beta gal activities using the o-nitrophenyl-beta-galactoside (ONPG) assay. Seventy five mutants were obtained out of more than 2 million colonies screened and showed increased beta-galactosidase activities compared with the wild-type strains. EMS gave a higher frequency of beta-gal overproducing mutants than MNNG for three of the four strains, S. thermophilus, B. breve, and B. longum, whereas the frequency of L. delbrueckii ssp. bulgaricus beta-gal mutants was similar with both mutagens. The highest beta-gal increases, when induced during growth in lactose, for mutants of each culture were 137% for L. delbrueckii ssp. bulgaricus; 104% for S. thermophilus; 70% for B. breve; and 222% for B. longum mutants. This food-grade classical approach has the ability to moderately increase beta-gal concentrations in probiotic cultures to improve their potential for treating the symptoms of lactose malabsorption in humans. PMID- 10821568 TI - Size distribution of fat globules in goat milk. AB - Milk from French-Alpine goats and Holstein cows was obtained from a bulk tank immediately prior to analyses. Fat globule size was determined by laser particle size analysis. Individual globules of fat in goat milk ranged from 0.73 to 8.58 microm in diameter. The average diameter of particles based on volume to surface area ratio (dvs) was 2.76 microm and was less than the mean (dvs) of 3.51 microm for bovine milk, in which fat globules ranged from 0.92 to 15.75 microm in diameter. The specific surface area of particles in caprine milk was 21,778 cm2/ml, whereas the specific surface area of particles in bovine milk was 17,117 cm2/ml. Ninety percent of the total particles found in goat milk were less than 5.21 microm in diameter, whereas 90% of the total particles in bovine milk were less than 6.42 microm based on the volume frequency distribution. Dissociation of casein micelles by urea in goat whole and skim milk caused larger dvs values due to the effect of fat particles and reduced the specific surface area in both milks because the total number of detectable particles in both whole and skim milk was reduced. PMID- 10821567 TI - Immediate effect of Lactobacillus acidophilus on the intestinal flora and fecal enzymes of rats and the in vitro inhibition of Escherichia coli in coculture. AB - The in vitro role of Lactobacillus acidophilus was investigated to explore the potential to inhibit coliforms. A threefold concentrated cell-free extract from L. acidophilus SBT2074 could efficiently inhibit most of the tested Gram-positive and Gram-negative bacteria. Among the three strains of L. acidophilus, SBT2062, SBT2071, and SBT2074, only L. acidophilus SBT2074 showed this inhibitory property. These three strains were also tested in coculture with Escherichia coli 3544 in skim milk medium. The fermentation could result in complete inhibition of E. coli in 36 h. Short-term administration of L. acidophilus SBT2074 in rats with and without E. coli resulted in significant inhibition of coliforms and anaerobes. The E. coli infected rats regained the normal flora in the presence of lactic acid bacteria. The fecal enzyme beta-glucuronidase activity was also decreased significantly when L. acidophilus SBT2074 was administered and was related to the decreased number of bacteria in the intestinal tract. The analysis of the small intestinal contents showed that the concentrations of coliforms in the duodenum, jejunum, and the ileum were significantly reduced by the administration of lactic acid bacteria. The effects are seen in a short period, suggesting that L. acidophilus SBT2074 fermentate may have clinical application for people suffering from gastrointestinal distress caused by coliforms. PMID- 10821569 TI - Generation of complement fragment C5a in milk is variable among cows. AB - The appearance of chemotactic fragments of complement at sites of infection is an important component of innate immunity. The contribution of C5a, the most biologically active complement fragment, to the recruitment of phagocytes in milk is not well defined, in particular the amount of C5a that is released in normal milk before inflammation. The generation of C5a in normal milk upon activation of complement by invading bacteria depends on the amount of available C5 and on the activity of the C3/C5-convertase of the alternative pathway. Concentrations of C5 were measured in one fore and one rear uninfected quarter of 19 Holstein cows. Values were consistent within cows, but widely dispersed among cows (0.19 to 1.94% blood concentration). C5 concentrations in milk were loosely related to concentrations in blood. By comparison, the range of milk concentrations of C3 (1.4 to 4.4%, mean 2.46 +/- 0.63% of blood concentration) was narrower. Two groups of six cows with high milk concentrations of C5 (cows H5: mean = 1.31%) and six cows with low milk concentrations of C5 (cow L5: mean = 0.21%) were constituted for further analysis of complement activation. There was a positive correlation between concentrations in milk of BSA and C5, but not between concentrations of BSA and C3. The activities of the C3- and C5-convertases were assessed through the deposition on complement-activating bacteria (Streptococcus agalactiae) of C3 and C5 fragments, respectively. The deposition of C3 was 1.7 fold higher, and the deposition of C5 was 2.75-fold higher in milk from H5 cows than in milk of L5 cows. Higher concentrations of C5 and better functioning of C5 convertase were mirrored by a much higher concentration of C5a in milk from H5 cows (12.30 ng/ml) than in milk of L5 cows (0.76 ng/ml) after activation of complement with zymosan. These results indicate that cows differed widely in their capacity to generate C5a in milk before inflammation, and that milk C5 concentrations were a primary limiting factor for C5a generation. Cows with the lowest milk concentrations of C5 are likely unable to use the complement system for the initial recruitment of leukocytes. PMID- 10821570 TI - Growth and morphogenesis of epithelial cell organoids from peripheral and medial mammary parenchyma of prepubertal heifers. AB - Collagen gel cell cultures were used to test the hypothesis that cells from peripheral parenchymal zones of the developing bovine mammary gland have greater proliferative and morphogenic potential than cells from the medial parenchymal mass. Tritiated thymidine was incorporated to assess cell proliferation. Dose responses for insulin-like growth factor one (IGF-I, 0 to 50 ng/ml) and transforming growth factor beta one (0 to 5 ng/ml) were determined for each cell population. Cells from the peripheral regions were two to three times more sensitive to the mitogenic action of IGF-I or serum. Transforming growth factor beta one concentrations of 12.5 to 500 pg/ml stimulated cell proliferation, but concentrations above 1 ng/ml were inhibitory. Peripheral cells showed a distinct biphasic response to addition of transforming growth factor beta one. Morphology of organoids was evaluated daily during culture and by histologic examination at the end of culture. Peripheral zone cells formed multi-layered cell structures that resembled native mammary parenchyma with serum or mammary gland extract. Our results indicate that epithelial cells of the peripheral and medial parenchymal zones in the ruminant mammary gland can differ in their proliferative and morphogenetic response to growth factor treatment in vitro. These differences likely reflect local tissue regulation necessary for sequential ductular and lobulo-alveolar development in vivo. PMID- 10821571 TI - Effects of photoperiodic manipulation during the dry period of dairy cows. AB - Thirty-four lactating Holstein cows were dried off 60 d prior to their expected calving date, paired by calving date, and randomly assigned to one of two photoperiod treatments: long-day photoperiod (n = 18; 16 h light: 8 h dark/d) or a short-day photoperiod (n = 16; 8 h light: 16 h dark/d) to determine if manipulation of photoperiod during the dry period would impact milk yield in the subsequent lactation. At parturition, calf weight, height, and length were measured and the cows were moved into ambient lighting conditions (November 1996 to January 1997 and November 1997 to January 1998, 39 degrees latitude) with the rest of the University of Maryland herd. After parturition, milk yield and composition were measured for 16 wk. Prolactin concentrations were higher by 11.7 ng/ml in the long-day photoperiod treatment group than for the cows on a short day photoperiod. During lactation cows previously exposed to a short-day photoperiod produced 3.2 kg/d more milk than long-day photoperiod cows. Energy corrected milk yield was greater in short-day photoperiod than in long-day photoperiod cows. Treatment did not affect calf growth, milk composition, or plasma insulin growth factor I concentrations. In conclusion, manipulation of photoperiod during the dry period of dairy cows may be a useful management tool for increasing milk yield in the subsequent lactation. PMID- 10821572 TI - The impact of somatotropin, milking frequency, and photoperiod on dairy farm nutrient flows. AB - Three technologies that increase milk production per cow and that are available to dairy producers are bovine somatotropin, three times daily milking, and extended daily photoperiod. Dairy herds fed according to National Research Council requirements were simulated to predict the impact of these technologies on N losses to manure and to water resources. Because Dairy Herd Improvement Association total lactation records (n = 93,080) revealed a positive linear relationship between 305-d milk production and calving interval, calving intervals were predicted to increase with the use of technologies and to result in a change in the ratio of lactating cows to growing heifers in a herd. Compared with a herd using no technologies, the use of bovine somatotropin, three times daily milking, or extended photoperiod were predicted to reduce herd N excretion per unit of milk by 7.8, 7.0, and 3.6%, respectively. When the use of all three technologies was simulated, N losses to manure were decreased by 15.7% when assuming calving interval increases from the technologies or 15.4% without accounting for calving interval increases. Reductions in feed N requirements and manure N losses with these three technologies were predicted to reduce environmental N loading by up to 16%. PMID- 10821573 TI - The response of altering the ratio of dietary protein to energy on growth, feed efficiency, and mammary development in rapidly growing prepubertal heifers. AB - This study was conducted to evaluate the effects of increasing the ratio of dietary protein to energy above National Research Council recommendations on average daily gain, feed efficiency, structural growth, and indirect measurements of mammary growth. Forty-five Holstein heifers were randomly assigned to either a low, medium, or high ratio of dietary crude protein (CP) to metabolizable energy of 46:1, 54:1, and 61:1 g/Mcal, respectively. The ratio of dietary protein to energy was altered by adjusting the concentration of CP with a similar amount of energy across all diets. Heifers were individually fed and began the treatment period at 200 kg of body weight and 28 wk of age. During the 3-wk adaptation period and 20-wk treatment period, all heifers were fed for a daily dry matter intake (DMI) of 2.45% of body weight. Body weight was monitored for two consecutive days each week and was used to adjust the dry matter offered on a weekly basis. The high versus low ratio of dietary protein to energy increased feed efficiency 6%, which resulted in larger heifers that were subsequently fed 3% more DMI over the course of the trial. The increased feed efficiency and DMI increased average daily gain by 9% for the high versus low ratio of dietary protein to energy. For the high versus low ratio of dietary protein to energy, hip width, hip height, wither height, and heart girth growth was increased 13, 16, 18, and 12%, respectively. The heifers fed the high ratio of dietary protein to energy had a lower rate of increased body condition score compared with the heifers fed the low ratio of dietary protein to energy. Teat length growth was used as an indirect measurement of mammary ductal development and was increased by 35 to 38% for heifers on the high versus low ratio of dietary protein to energy. Feeding dietary ratios of protein to energy above NRC recommendations improved feed efficiency and increased average daily gain, structural growth, and mammary development while decreasing body condition scores in heifers between 28 and 48 wk of age. PMID- 10821574 TI - Duration of regrowth of ryegrass (Lolium perenne) effects on grazing behavior, intake, rumen fill, and fermentation of lactating dairy cows. AB - The relative importance of duration of sward regrowth and fill and fermentation in the rumen on the control of grazing time and intake rate during the first grazing session of the day was studied. Four lactating dairy cows were allowed to graze ryegrass (Lolium perenne) swards, with five different regrowth periods after mowing (6, 9, 16, 22, and 30 d). The cows were allowed to graze until they stopped voluntarily (cessation of grazing activity for at least 15 min). Before and after grazing the rumen contents were evacuated, weighed, sampled, and returned to the animals. Samples of rumen liquid were taken immediately before rumen evacuation and approximately 30, 60, 120, and 240 min after the grazing session was finished. Grazing time and intake rate did not follow a significant trend with period of regrowth. Bite rate did not change significantly with duration of regrowth with cows exhibiting high rates of biting for all the sward conditions. Rumen pools sizes of dry matter, neutral detergent fiber, and volatile fatty acids (VFA), measured after grazing, increased significantly with days of sward regrowth, even though the changes over days were small. Concentration of VFA followed a significant quadratic trend with a maximum concentration observed at approximately 110 min after cessation of grazing. In this study, rumen fill, VFA (either total or major components), ammonia, pH, and osmotic pressure as individual variables were not correlated with grazing time or dry matter intake. PMID- 10821575 TI - Lysine metabolism by the mammary gland of lactating goats at two stages of lactation. AB - An arteriovenous kinetics technique was used to monitor mammary gland lysine and protein metabolism in goats (n = 4) at two stages of lactation (80 +/- 17 vs. 233 +/- 14 DIM) in response to an i.v. infusion of lysine (Lys) plus methionine (Met). At each stage of lactation [2-15N] and [1-13C; 6,6-2H2] Lys kinetics were performed on the last day of 5-d i.v. infusion of saline followed by Lys (370 mg/h) plus Met (84 mg/h, LM). Milk and protein yields and dry matter intake were higher in early than in late lactation, but LM infusion did not affect these variables. Regardless of stage of lactation, the absolute and fractional oxidation rates of Lys by the mammary gland increased in response to LM infusion. When corrected for Lys oxidation, net uptake of Lys by the gland was less than milk protein Lys secretion. However, correction for the contribution of peptides (15.8%) to Lys uptake brought net Lys uptake close into balance with milk Lys secretion. The present data suggests that when Lys is in excess of requirements, the mammary gland appears to dispose of the extra supply via the oxidative mechanism. PMID- 10821576 TI - Effects of grain processing and bovine somatotropin on metabolism and ovarian activity of dairy cows during early lactation. AB - This study compared the effects of exogenous bovine somatotropin (bST) on the metabolism and ovarian activity of cows fed diets differing in ruminally degradable starch. Twenty-four multiparous and eight primiparous Holstein cows in early lactation were divided into four groups and fed diets containing 39% grain as steam-flaked sorghum or steam-rolled corn with or without bST for 90 d in a 2 x 2 factorial design. Flaked sorghum improved energy status of cows during early lactation, tending to increase plasma glucose and insulin. Administration of bST decreased plasma urea nitrogen and increased nonesterified fatty acids (NEFA). Plasma levels of beta-hydroxybutyrate (BHBA) and hepatic concentrations of triglycerides were not altered by treatments. Temporal changes in plasma glucose, urea nitrogen, NEFA, and BHBA were detected in a quadratic manner and insulin increased linearly with time, but treatments did not affect postpartum changes in these metabolites. There were greater decreases in body weight and net energy balance in cows on bST during the first 7 wk of treatment. Cows receiving bST took longer to reach the nadir of negative energy balance, and bST tended to delay the period to reach a positive energy balance. Follicular populations and incidence of cystic ovaries were not affected by treatments, but cows receiving bST had fewer double ovulations. Flaked sorghum increased plasma progesterone during the early luteal phase of the first two postpartum estrous cycles. Feeding more ruminally degradable starch improved the energy status and luteal activity of cows in early lactation. PMID- 10821578 TI - Milk production, reproductive performance, and fecal excretion of phosphorus by dairy cows fed three amounts of phosphorus. AB - Milk production was measured and phosphorus (P) excretion in feces was estimated in dairy cows fed three amounts of P. A basal diet was formulated to contain 0.31% P (DM basis). Sodium monophosphate replaced corn in the basal diet to give two additional diets containing 0.40 and 0.49% P. The diets were fed to eight, nine, and nine multiparous Holsteins from the beginning to the end of lactation. Milk yields for the 308-d lactation were 10,790, 11,226, and 11,134 kg for the three treatments, respectively. The lowest milk yield resulted from decreased milk production during late lactation with the 0.31% P group. Reproductive performance of the cows was not related to dietary P content. Fecal P concentration, determined in wk 2, 4, 6, 8, 23, and 40 of lactation, increased as dietary P intake was increased. Cows fed the lowest P diet conserved P by minimizing P excretion in feces and urine, whereas cows in the other two treatments excreted more P through these routes. A reduction in dietary P from 0.49 to 0.40% reduced fecal P excretion by 23%. Apparent P digestibilities of less than 40% are indicative of surplus dietary P. Feeding 0.40% P appeared sufficient to maintain P balance and the level of milk production achieved in this experiment. An example is given which illustrates the relationship between dietary and fecal P. PMID- 10821577 TI - Conjugated linoleic acid (CLA) content of milk from cows offered diets rich in linoleic and linolenic acid. AB - Two experiments were conducted to determine the conjugated linoleic acid (CLA) content of milk from cows offered diets rich in linoleic and linolenic acid. In experiment 1, 36 cows were assigned to a control and five treatment groups. Cows in the control group received a diet containing 51% forage and 49% grain on a dry matter basis. In the treatment group, grain was partly replaced by either 18% raw cracked soybeans, 18% roasted cracked soybeans, 3.6% soybean oil, 2.2% linseed oil, or 4.4% linseed oil. Experimental diets were fed for 5 wk. Average CLA contents in milk fat from wk 2 through 5 were 0.39% in control and 0.37, 0.77, 2.10, 1.58, and 1.63% of total fatty acids in the raw soybean, roasted soybeans, soybean oil, 2.2% linseed oil, and 4.4% linseed oil treatments, respectively. In experiment 2, 36 cows were assigned to a control and 5 treatment groups. Cows in the control group received a diet containing 55% forage and 45% grain. In the treatment groups, grain was partly replaced by soybean oil at 0.5, 1.0, 2.0, 4.0, or by linseed oil at 1.0% of the dietary dry matter. Experimental diets were fed for 5 wk. Average CLA contents in milk fat from wk 2 through 5 were 0.50% in control and 0.75, 0.76, 1.45, 2.08, and 0.73% of total fatty acids in 0.5, 1.0, 2.0, 4.0 soybean oil and 1.0% linseed oil treatments, respectively. Diets rich in linoleic or linolenic acid can increase CLA content of milk when dietary oil is accessible to the rumen microorganisms. PMID- 10821579 TI - Milk production during the complete lactation of dairy cows fed diets containing different amounts of protein. AB - Milk production response to four different amounts of protein supplementation was measured in a complete lactation study utilizing 58 multiparous Holstein cows treated with bovine somatotropin. The four treatments were as follows (the first number is the dietary crude protein content (% of dry matter) during the first 16 wk of lactation and the second number is the protein content for wk 17 to 44 of lactation): 15.4-16.0, 17.4-16.0, 17.4-17.9, and 19.3-17.9. Diets were formulated to maximize the supply of amino acids to the intestine. High moisture ear corn was finely ground to promote ruminal fermentation and microbial protein synthesis, and roasted soybeans and expeller processed soybean meal were used as the protein supplements to provide relatively high amounts of rumen-undegradable protein. Alfalfa silage and corn silage (3:2) provided the forage. Milk production for the 308-d lactation for each of the treatment groups was 10,056, 10,831, 11,095, and 11,132 kg. Cows of this production level fed diets similar to those used in this experiment benefit from dietary protein of approximately 17.5% during the first 30 wk of lactation. A reduction in dietary protein to 16% can be made around wk 30 of lactation. This amount of dietary protein should, with prevailing feed prices, be compatible with maximum profit and a moderate amount of nitrogen excretion to the environment. PMID- 10821580 TI - Milk production and reproductive performance of dairy cows fed two concentrations of phosphorus for two years. AB - The performance of lactating Holstein cows in response to P supplementation was determined in a 2-yr study. Each year included confinement feeding for approximately the first two-thirds of lactation and grazing for the remaining one third of lactation. In yr 1, 42 cows were assigned at calving to a low or high P diet within parity. Fourteen cows from the low P group and 16 cows from the high P group continued with their treatments for a second year. Also in the second year, 12 new cows were included in the low P group and 11 in the high P group. Thus, a total of 95 lactations with 65 cows were used in the trial, and 30 of the cows were used in both years. The dietary P was 0.38 and 0.48% during confinement feeding and approximately 0.31 and 0.44% during grazing for the low and high P treatments (dry basis). When all cows were used to obtain treatment means, milk yield for 308 d of lactation was 9131 and 8860 kg in yr 1, and 9864 and 9898 kg in yr 2 for the low P and high P groups, respectively. Blood serum inorganic P tended to be slightly lower for the low P than for the high P group during most of lactation; all concentrations (5.6 to 7.4 mg/dl) were within normal ranges. Reproductive measures were similar between groups in both years. When just the cows completing two lactations (N = 30) were evaluated, milk yield was 9072 and 8780 kg in yr 1 and 11,457 and 11,358 kg in yr 2 for the low P and high P treatments, respectively. Reducing dietary P from 0.48 to 0.38% for 2 yr did not impair milk production or reproductive performance. PMID- 10821581 TI - Relationships between mastitis and functional longevity in Danish Black and White dairy cattle estimated using survival analysis. AB - The relationship between mastitis and functional longevity was assessed with survival analysis on data of Danish Black and White dairy cows. Different methods of including the effect of mastitis treatment on the culling decision by a farmer in the model were compared. The model in which mastitis treatment was assumed to have an effect on functional longevity until the end of the lactation had the highest likelihood, and the model in which mastitis treatment had an effect for only a short period had the lowest likelihood. A cow with mastitis had 1.69 times greater risk of being culled than did a healthy herdmate with all other effects being the same. A model without mastitis treatment was used to predict transmitting abilities of bulls for risk of being culled, based on longevity records of their daughters, and was expressed in terms of risk of being culled. The correlation between the risk of being culled and the national evaluations of the bulls for mastitis resistance was approximately -0.4, indicating that resistance against mastitis was genetically correlated with a lower risk of being culled and, thus, a longer functional length of productive life. PMID- 10821582 TI - Genetic parameters for reproductive traits of Holstein cattle in California and Minnesota. AB - Genetic parameters for five reproductive traits were estimated using data from 51,528 Holstein cows that were inseminated from April to September 1998 in 1717 herds in California and Minnesota. Nonreturn rate and veterinary-confirmed pregnancy rate at 60 and 90 d after insemination were evaluated using linear and threshold models, including an additive genetic effect for the cow being inseminated and a random environmental effect for the service bull. Interval from calving to first insemination was evaluated using a linear model, including an additive genetic effect for the cow being inseminated. Linear model heritability estimates for 60-d (90-d) nonreturn rate were 1.4% (1.5%) in California and 4.1% (2.7%) in Minnesota. Corresponding estimates for 60-d (90-d) confirmed pregnancy rate were 1.4% (2.3%) in California and 1.0% (2.0%) in Minnesota; the proportion of cows with veterinary data available 60 d after breeding was 86% in California and 55% in Minnesota. Threshold model heritability estimates were slightly higher than linear model estimates in California but were lower in Minnesota, presumably because 25% of the herd-season classes in Minnesota contained either all successes or all failures. Linear model repeatability estimates for the service bull effect on 60-d (90-d) nonreturn rate were 0.5% (0.4%) in California and 0.3% (0.3%) in Minnesota. Corresponding estimates for 60-d (90-d) confirmed pregnancy rates were 0.6% (0.2%) in California and 0.1% (0.4%) in Minnesota. Threshold model estimates were slightly higher than linear model estimates in both states. Heritability estimates for the interval from calving to first insemination were 5.8% in California and 6.1% in Minnesota. Despite the low parameter estimates, variation was present among animals, and it should be possible to identify sires that possess superior or inferior reproductive characteristics. PMID- 10821583 TI - Retrospective analysis of the accuracy of conversion equations and multiple trait, across-country evaluations of Holstein bulls used internationally. AB - In 1995, the multiple-trait across country genetic evaluation procedure replaced regression-based conversion equations as the preferred method for international genetic comparisons of dairy bulls. In the present study, February 1999 estimated breeding values of 632 foreign Holstein bulls that were used in Canada, Germany, Italy, The Netherlands, Sweden, and the US were compared with January 1995 predictions from home country data only. January 1995 predicted breeding values for each importing country were calculated using three methods: the multiple trait, across-country evaluation procedure; conversion equations based on the multiple-trait, across-country evaluations; and conversion equations based on the Wilmink method. Mean correlations between 1999 estimated breeding values in the importing countries and 1995 predictions from international data were from 0.76 to 0.81 for all methods. The multiple-trait, across-country evaluation procedure is expected to lead to selection of different bulls, because bulls were allowed to be ranked differently in each country, but no significant increase in accuracy of selection was observed. The lack of improvement in accuracy of prediction was most likely due to limitations in data structure. International genetic comparisons are largely driven by data from a relatively small number of evaluated bulls with exported semen. Data from siblings and more distant relatives provide only weak, indirect genetic links between countries, and inclusion of such data seems to provide a minimal improvement in accuracy. Limitations in data structure might be alleviated by methods that define environments by climate or management factors rather than country borders. PMID- 10821584 TI - A link function approach to model heterogeneity of residual variances over time in lactation curve analyses. AB - Several studies with test-day models for the lactation curve show heterogeneity of residual variance over time. The most common approach is to divide the lactation length into subclasses, assuming homogeneity within these classes and heterogeneity between them. The main drawbacks of this approach are that it can lead to many parameters being estimated and that classes have to be arbitrarily defined, whereas the residual variance changes continuously over time. A methodology that overcomes these drawbacks is proposed here. A structural model on the residual variance is assumed in which the covariates are parametric functions of time. In this model, only a few parameters need to be estimated, and the residual variance is then a continuous function of time. The analysis of a sample data set illustrates this methodology. PMID- 10821585 TI - Time series autoregressive integrated moving average modeling of test-day milk yields of dairy ewes. AB - Monthly test-day milk yields of 1200 dairy Sarda ewes were analyzed by time series methods. Autocorrelation functions were calculated for lactations within parity classes and altitude of location of flocks. Spectral analysis of the successions of data was developed by Fourier transformation, and different Box Jenkins autoregressive integrated moving average models were fitted. The separation of deterministic and stochastic components highlighted the autoregressive feature of milk production pattern. The forecasting power of autoregressive integrated moving average models was tested by predicting total milk production for a standardized lactation length of 225 d from only a few test day records. Results indicated a greater forecasting capacity in comparison with standard methods and suggested further development of time-series analysis for studying lactation curves with more sophisticated methods, such as wavelet decomposition and neural network models. PMID- 10821586 TI - Development of a cost analysis spreadsheet for calculating the costs to raise a replacement dairy heifer. AB - Dairy operations have a variety of resources and objectives, such that the most economical method of obtaining replacement heifers is only determined by individual analysis of costs. The objective of this study was the development of a cost analysis spreadsheet and validation of that spreadsheet on milking and custom heifer operations throughout Pennsylvania. A cost analysis spreadsheet was developed with an Excel '97 Microsoft file. The spreadsheet estimated the costs to raise a replacement heifer by specific age classes for feed, labor, health, reproduction, bedding, facilities, equipment, mortality, and interest costs. The simplistic and broad-based nature of the spreadsheet was a key component in the spreadsheet's flexibility to estimate costs for a variety of operational objectives, feeding management, housing systems, and labor management. A convenience sample of 16 milking operations and 14 custom heifer operations was evaluated to validate the cost analysis spreadsheet. Results from the validation are discussed to highlight the success and performance of the cost analysis spreadsheet. The average total cost to raise a replacement heifer for this data set was $1124.06 and $1019.20 for milking and custom heifer operations, respectively. Feed costs contributed 60.3 and 64.0% of the average total cost for milking and custom heifer operations, respectively. While no two operations are alike, individual operations possessing the ability to address costs to raise a replacement heifer can utilize critical information that can be used to improve operation profitability. PMID- 10821587 TI - Liner wall movement and vacuum measured by data acquisition. AB - The objective of this study was to measure liner wall movement and define the characteristics of pulsation. The cyclic opening and closing of the milking machine liner is called "Pulsation." The liner opening and closing is a consequence of the differential pressures across the walls of the liner barrel. Liner wall movement was measured in the laboratory and during milking with an analogue laser sensor and a laptop computer equipped with a data acquisition system. Measurements were made in 10-s segments. The first and last cycles of 10 pulsations were dropped from each segment, and the average of eight cycles was used for final analysis. Opening and closing phases of liner wall movement were characterized at the intersection of 1 mm less than fully open and 1 mm greater than fully closed. The 'D' phase of the liner was 407 ms, in contrast to the 'd' phase of the chamber vacuum of 259 ms with an artificial teat. Time of liner opening averaged 87 ms with an artificial teat and 98 ms during milking. Liner closure time was 23 ms with an artificial teat and 42 ms during milking. Liner ratio was 57:43, with a chamber vacuum ratio of 65:35 using an artificial teat. Liner ratio was 58.2:41.8 with a chamber ratio of 64.5:35.5 during milking. This technology can be used to improve liner design and enhance milking performance. PMID- 10821588 TI - Theoretical basis and computational methods for different test-day genetic evaluation methods. AB - In test-day (TD) models, records from individual test days are used to determine lactation production instead of aggregating records. Test-day models have recently gained considerable interest because they are more flexible in handling records from different recording schemes. Compared with only using records of complete lactations, they can reduce the generation interval through frequent genetic evaluations with the latest data. Test-day models can predict total production more accurately by accounting for time-dependent environmental effects. Test-day models may be separated into three groups: First, two-step models under which corrections are carried out at TD level and subsequently corrected TD records are processed in an aggregated form as lactation records. Second, fixed regression models assume that TD records within a lactation are repeated records. Because yields in the course of the lactation follow a curvilinear pattern, this curve can be considered by using suitable covariates. Third, random regression models additionally define the animal's genetic effect by using regression coefficients and allowing for covariances among them. The difference between random regression and fixed regression models is that the genetic merit of an individual is allowed to differ in the course of the lactation in random regression models. Random regressions are related to the approach of defining covariance functions for longitudinal data. Computationally, TD models are very demanding. For evaluations on a national scale, the size of the equation system can go to hundreds of millions of equations, depending on the size of the database and the specific model defined. PMID- 10821589 TI - Strategies for estimating the parameters needed for different test-day models. AB - Currently, most analyses of parameters in test-day models involve two types of models: random regression, where various functions describe variability of (co)variances with regard to days in milk, and multiple traits, where observations in adjacent days in milk are treated as one trait. The methodologies used for estimation of parameters included Bayesian via Gibbs sampling, and REML in the form of derivative-free, expectation-maximization, or average-information algorithms. The first method is simpler and uses less memory but may need many rounds to produce posterior samples. In REML, however, the stopping point is well established. Because of computing limitations, the largest estimations of parameters were on fewer than 20,000 animals. The magnitude and pattern of heritabilities varied widely, which could be caused by simplifications in the model, overparameterization, small sample size, and unrepresentative samples. Patterns of heritability differ among random regression and multiple-trait models. Accurate parameters for large multi-trait random regression models may be difficult to obtain at the present time. Parameters that are sufficiently accurate in practice may be obtained outside the complete prediction model by a constructive approach, where parameters averaged over the lactation would be combined with several typical curves for (co)variances for days in milk. Obtained parameters could be used for any model, and could also aid in comparison of models. PMID- 10821590 TI - Experience with a test-day model. AB - The Canadian Test-Day Model is a 12-trait random regression animal model in which traits are milk, fat, and protein test-day yields, and somatic cell scores on test days within each of first three lactations. Test-day records from later lactations are not used. Random regressions (genetic and permanent environmental) were based on Wilmink's three parameter function that includes an intercept, regression on days in milk, and regression on an exponential function to the power -0.05 times days in milk. The model was applied to over 22 million test-day records of over 1.4 million cows in seven dairy breeds for cows first calving since 1988. A theoretical comparison of test-day model to 305-d complete lactation animal model is given. Each animal in an analysis receives 36 additive genetic solutions (12 traits by three regression coefficients), and these are combined to give one estimated breeding value (EBV) for each of milk, fat, and protein yields, average daily somatic cell score and milk yield persistency (for bulls only). Correlation of yield EBV with previous 305-d lactation model EBV for bulls was 0.97 and for cows was 0.93 (Holsteins). A question is whether EBV for yield traits for each lactation should be combined into one overall EBV, and if so, what method to combine them. Implementation required development of new methods for approximation of reliabilities of EBV, inclusion of cows without test day records in analysis, but which were still alive and had progeny with test-day records, adjustments for heterogeneous herd-test date variances, and international comparisons. Efforts to inform the dairy industry about changes in EBV due to the model and recovering information needed to explain changes in specific animals' EBV are significant challenges. The Canadian dairy industry will require a year or more to become comfortable with the test-day model and to realize the impact it could have on selection decisions. PMID- 10821591 TI - Consequences of selection for milk yield from a geneticist's viewpoint. AB - The annual genetic trend for milk yield of Holsteins in the United States has accelerated with time and had means of 37 kg during the 1960s, 79 kg during the 1970s, 102 kg during the 1980s, and 116 kg from 1990 to 1996. Selection programs of the dairy cattle breeding firms in the United States have become more selective and effective with time, and selection goals continue to place major emphasis on yield traits, which clearly impact profitability of dairying. Traits other than yield are also included in selection goals of the industry. Type traits, especially those related to udder conformation, body size, and angularity have been included in selection programs and have altered the appearance and physiological functions of Holstein cows. Selection programs have continued to increase the body size of Holsteins despite mounting evidence that smaller cows have advantages for survival and efficiency. Favorable emphasis on cows that appear sharper might result in cows that are more prone to metabolic problems. The high intensity of current selection in the United States has brought about a rapid increase in genetic relationships among animals. Increased relationships will inevitably result in undesirable levels of inbreeding in the commercial cow population unless dairy producers turn to crossbreeding. PMID- 10821592 TI - Selection for milk production from a lactation biology viewpoint. AB - The success of selection for increased milk production in dairy cows is apparent. Certainly, many herds now have average production levels that would have only been associated with the best producers in the herd 30 yr ago. There are, of course, many reasons for this success. Among these are improvements in genetic selection methods and associated use of artificial insemination, better fulfillment of nutritional needs and diet formulation, and careful attention to mastitis control and milking management. Development of new management tools (i.e., bovine somatotropin, improved crops, estrus detection devices, estrus synchronization, monitoring of individual animal performance, and disease prevention) should not be forgotten. Although many aspects of a dairy operation determine overall performance and profitability, the focus of this paper is the udder. Information indicates that both the structure and function of the bovine mammary gland have been directly impacted by long-term selection for increased milk production but improved functionality may have been more important. This review also considered studies that attempt to develop techniques and measurements for possible selection of genetically superior animals including measurement of circulating hormones and direct assay of mammary tissue function. PMID- 10821593 TI - The next $120,000: a case study to illustrate analysis of alternative farm investments in fixed assets. AB - Dairy scientists specializing in the area of farm management are increasingly involved in analysis of farm investments in fixed assets. There have been instances where the wrong procedures were used to assess investments in fixed assets, leading to erroneous and possibly disastrous conclusions. A detailed case study of a dairy farm facing the decision of where best to invest an unexpected $120,000 windfall is used to illustrate the various facets of financial analysis. Indicators of profitability, liquidity, solvency, repayment capacity, and financial efficiency are explained and applied to the farm case to produce a detailed analysis of the current financial position of the firm. Long-range budgets of four alternate investment options and their impact on all financial indicators are presented. The four options are: 1) to pay down debt, 2) to purchase an additional 100 cows, 3) to install automatic milk yield recording in the parlor, and 4) to build new heifer facilities. All four investments are profitable. Therefore, an analysis limited to profitability indicators would conclude that any of the four options is a good investment. However, liquidity and financial efficiency issues showed that the option of purchasing 100 cows is far superior to the three others. We conclude that a complete and thorough financial analysis is required to evaluate the impact of long-run investments in fixed assets. PMID- 10821594 TI - Effect of coinfection with STDs and of STD treatment on HIV shedding in genital tract secretions: systematic review and data synthesis. AB - OBJECTIVE: To determine whether coinfection with sexually transmitted diseases (STD) increases HIV shedding in genital-tract secretions, and whether STD treatment reduces this shedding. DESIGN: Systematic review and data synthesis of cross-sectional and cohort studies meeting predefined quality criteria. MAIN OUTCOME MEASURES: Proportion of patients with and without a STD who had detectable HIV in genital secretions, HIV load in genital secretions, or change following STD treatment. RESULTS: Of 48 identified studies, three cross-sectional and three cohort studies were included. HIV was detected significantly more frequently in participants infected with Neisseria gonorrhoeae (125 of 309 participants, 41%) than in those without N gonorrhoeae infection (311 of 988 participants, 32%; P = 0.004). HIV was not significantly more frequently detected in persons infected with Chlamydia trachomatis (28 of 67 participants, 42%) than in those without C trachomatis infection (375 of 1149 participants, 33%; P = 0.13). Median HIV load reported in only one study was greater in men with urethritis (12.4 x 104 versus 1.51 x 104 copies/ml; P = 0.04). In the only cohort study in which this could be fully assessed, treatment of women with any STD reduced the proportion of those with detectable HIV from 39% to 29% (P = 0.05), whereas this proportion remained stable among controls (15-17%). A second cohort study reported fully on HIV load; among men with urethritis, viral load fell from 12.4 to 4.12 x 104 copies/ml 2 weeks posttreatment, whereas viral load remained stable in those without urethritis. CONCLUSION: Few high-quality studies were found. HIV is detected moderately more frequently in genital secretions of men and women with a STD, and HIV load is substantially increased among men with urethritis. Successful STD treatment reduces both of these parameters, but not to control levels. More high-quality studies are needed to explore this important relationship further. PMID- 10821595 TI - Reexamining the prevalence of Chlamydia trachomatis infection among gay men with urethritis: implications for STD policy and HIV prevention activities. AB - BACKGROUND: Evidence of an STD-HIV interaction and the availability of noninvasive urine-based screening tests have resulted in an increased focus on chlamydial infections in men. GOAL: To evaluate the prevalence of chlamydial infections among men with urethritis at the San Francisco City Clinic (SFCC). STUDY DESIGN: In 1997, male SFCC patients diagnosed with urethritis were tested for chlamydia using urine-based ligase chain reaction and for gonorrhea using urethral culture. RESULTS: Gonorrhea was identified in 45% of men who have sex with men (MSM) versus 26% of men who have sex with women (MSW). Among men with gonorrhea, chlamydia coinfection was found among 15.2% of MSM and 8.4% of MSW. Among men with nongonococcal urethritis, 18% and 20% of MSM and MSW had chlamydial infection, respectively. Young age was associated with chlamydial infection in MSM. CONCLUSION: After a period of low chlamydial infection rates in MSM during the pre-AIDS era, infection rates are increasing among this population. SFCC's revised clinical practice guidelines include chlamydia testing of MSM with urethritis. PMID- 10821596 TI - Histologic development of cervical ectopy: relationship to reproductive hormones. AB - BACKGROUND AND OBJECTIVES: Cervical ectopy has been proposed as a risk factor for chlamydial infection, HIV, and other sexually transmitted diseases (STDs). Ectopy is a histologic phenomenon resulting from a complex interplay of morphologic and hormonal processes. Reproductive hormones influence the production of ectopy during late fetal life, puberty, pregnancy, and with use of oral contraceptives. Ectopy is modified over time by squamous metaplasia and epitheliazation, low pH, trauma, and possibly by cervical infection. STUDY DESIGN: The authors review the histologic development of ectopy from late fetal life to menopause to understand STD risk. RESULTS: Ectopy is often assessed and quantified by direct, unaided observation during speculum examinations. This method may result in inaccurate estimates and misclassification with the transformation zone. CONCLUSION: Valid, reproducible measures of ectopy are necessary for epidemiologic studies to examine the impact of exogenous hormonal contraceptives on the development of ectopy and on potential infection risk. PMID- 10821597 TI - HIV-1 infection-associated risk factors among sexually transmitted disease patients in Athens, Greece: 1990 to 1996. AB - OBJECTIVE: To determine trends in HIV seroprevalence and related risk factors among patients with sexually transmitted diseases (STDs) and to report the respective epidemiologic history characteristics. METHODS: A cross-sectional seroepidemiologic study conducted from 1990 to 1996 among 5,669 symptomatic STD cases was carried out. RESULTS: The overall HIV test acceptance was 98.9%, and 1.2% patients (n = 66) were seropositive. Highest rates were detected among those who were born or resided in Sub-Saharan Africa. Seropositivity fluctuates significantly by age, and is excessive in persons 45 years and older (2.6%). A significant decreasing trend in STD incidence and HIV seroprevalence among patients younger than 25 years was detected. Male homosexuals and bisexuals (MSM) exhibited the highest overall rate of infection (5.8%) followed by intravenous drug users (2%). Highly promiscuous STD patients (ie, those who had more than 10 partners during the past 6 months) presented a significantly increased HIV seroprevalence when compared with patients of the same sexual orientation. STD patients infected with HIV mostly belonged to notable risk categories of AIDS (men who have sex with men, 72.7%). Awareness of serostatus was low (13.6%). In male patients, the HIV seropositivity rate was significantly higher among early syphilis and proctitis cases, whereas in females this higher rate occurred with herpes genitalis. CONCLUSIONS: Promiscuity and sexual orientation significantly influence the seroprevalence rate. Exposure to HIV remained stable despite the above declining time trends, which implies the need for additional preventive interventions targeted to the real health and illness behavior of the partner. PMID- 10821598 TI - Premarket evaluation of the POCkit HSV-2 type-specific serologic test in culture documented cases of genital herpes simplex virus type 2 [see comment]. AB - BACKGROUND AND OBJECTIVES: The genital herpes epidemic continues, in part, because patients with subclinical or atypical presentations cannot be identified by most herpes simplex virus (HSV) antibody tests. A new product, POCkit HSV-2, has been developed to rapidly and accurately detect antibodies to HSV type 2 (HSV 2) in capillary blood or serum. GOAL: Sera from patients with culture-documented genital or oral herpes were tested to determine the sensitivity and specificity of the POCkit HSV-2 rapid point-of-care antibody test (Diagnology, Belfast, Northern Ireland). STUDY DESIGN: Sera from 50 patients with culture-documented HSV type 1 (9 oral, 41 genital) and from 253 patients with genital HSV-2 were tested by POCkit HSV-2 for HSV-2 antibodies. Each subject had a positive culture for HSV within 6 months of serum collection. Sera were preselected to include only those that were seropositive to the respective virus subtype by University of Washington Western blot. RESULTS: Compared with viral culture and Western blot analysis, sensitivity of the POCkit HSV-2 test for HSV-2 antibody was 96%; specificity was 98%. CONCLUSION: This test provides rapid, accurate identification of HSV-2 antibody in subjects with established HSV infections. PMID- 10821599 TI - Testing for herpes simplex virus type 2: full-steam ahead? PMID- 10821600 TI - Sexually transmitted infections in women with disabilities: diagnosis, treatment, and prevention: a review. AB - BACKGROUND AND OBJECTIVES: Women with disabilities can and do contract sexually transmitted diseases (STDs). However, because of access barriers, attitudinal misconceptions, and lack of awareness of their risk status, these infections may go under or undiagnosed. GOAL: To address issues regarding the treatment of women with disabilities and to suggest ways in which the patient and the provider can work together for a positive outcome. STUDY DESIGN: A literature review was performed using recent textbooks, publications, and medical journals to collect information regarding the health behaviors of women with disabilities and sexually transmitted disease symptom manifestations, which may differ in this population. The Physicians Desk Reference was consulted to assemble information on medical selection. RESULTS: The treatment of STDs in women with disabling conditions presents diagnostic and therapeutic challenges to clinicians, because symptoms may be confusing and may mimic manifestations of underlying disorders. Women with spinal cord injury may be at risk for the development of autonomic activation as a sign of STDs. CONCLUSION: To enhance compliance with medication regiments, the limitations of the patient should be considered. Furthermore, women with disabilities are at high risk for sexual abuse; therefore, the presence of an STD may be of special concern. Developing good communication with the patient will enable the clinician to work with her to sort out symptoms, design therapeutic regiments, and to help protect her from abuse. PMID- 10821602 TI - Incidence and predictors of reinfection with Trichomonas vaginalis in HIV infected women. AB - BACKGROUND AND OBJECTIVES: The presence of sexually transmitted infections (STIs) may facilitate transmission of HIV to uninfected partners. GOAL: To describe the incidence of reinfection with Trichomonas vaginalis in HIV-infected women and to assess predictors of reinfection. STUDY DESIGN: A retrospective cohort study using data abstracted from medical records of HIV-infected women with at least one diagnosis of trichomoniasis. RESULTS: Approximately one third (36%) of the study population was reinfected with T vaginalis during the follow-up period, with an incidence of 16.4 reinfections per 100 person years. Significant predictors of reinfection included history of another STI (hazard ratio, 1.52; 95% CI, 1.08-2.14) and becoming pregnant during the follow-up period (hazard ratio, 0.59; 95% CI, 0.39-0.87). CONCLUSIONS: There is a high rate of reinfection with T vaginalis in HIV-infected women. Further research that includes information on sexual partners should be conducted to better describe the reinfection patterns of trichomoniasis. PMID- 10821601 TI - Neisseria gonorrhoeae MS11mkC opacity protein expression in vitro and during human volunteer infectivity studies. AB - BACKGROUND AND OBJECTIVES: Neisseria gonorrhoeae MS11mkC harbors 11 independently expressed opacity (Opa) protein genes with distinct in vitro expression frequencies. In experimental infections in which human male volunteers were inoculated with transparent (Opa), piliated (P+) strains, the authors associate onset of symptoms with recovery of opaque (Opa+) gonococci. GOALS: In vitro and recovered (Opa) protein expression rates were compared to determine if the human host influences Opa expression. STUDY DESIGN: Opa expression was determined using Western immunoblot analysis; Opa sizes were determined using a scanning densitometer. RESULTS: Seven of 10 Opa proteins were identified in gonococci recovered from all of the volunteers at frequencies consistent with in vitro results (Opa C, 29.5 kDa; Opa K, 30 kDa; Opa G, 31 kDa; Opa I, 32 kDa; Opa J, 33 kDa; Opa D, 34 kDa; and Opa H, 37 kDa) (P > or = 0.01, Fisher exact test). Opa B (30.5 kDa) was identified at lower than expected frequencies, whereas Opa E (31.2) and F (31.5) were identified at higher' than expected frequencies. When recovered gonococci were reanalyzed for in vitro expression frequencies, they were consistent with preinfection frequencies. CONCLUSIONS: The host may influence the prevalence of some Opa proteins. PMID- 10821603 TI - Risk factors for pelvic inflammatory disease in inner-city adolescents. AB - OBJECTIVE: To determine risk factors associated with pelvic inflammatory disease (PID) among inner-city adolescents. STUDY DESIGN: A case-control study was performed from 1994 to 1997 in an inner-city hospital. METHODS: Seventy-one adolescent girls diagnosed with PID and 52 sexually active adolescents girls without PID participated in a confidential face-to-face interview using a questionnaire about risk behaviors. Established criteria were used for the diagnosis of PID. Data were analyzed using t tests, chi-square tests, and stepwise logistic regression. RESULTS: Persons with PID were significantly more likely to show younger age at first intercourse, older sex partners, involvement with a child protection agency, prior suicide attempt(s), consumption of alcohol before last sex, and a current Chlamydia trachomatis infection. There were no significant differences between the two groups regarding number of lifetime sex partners, condom use, rape, syphilis, prior PID, hepatitis B, hepatitis C, or HIV infection. CONCLUSIONS: Not previously noted in the literature are the association of PID with older sex partners, prior involvement in a child protection agency, and a prior suicide attempt. Confirming prior studies are the association of PID with earlier age at first sex, alcohol use, and C trachomatis infection. PMID- 10821604 TI - Seroprevalence of herpes simplex virus type 2 infection among attendees of a sexually transmitted disease clinic in Italy. Italian Herpes Forum. AB - BACKGROUND AND OBJECTIVES: An increased prevalence of herpes simplex virus type 2 (HSV-2) infection has been recently observed in industrialized countries. GOAL: To determine HSV-2 seroprevalence in a high-risk population in Italy. STUDY DESIGN: A cross-sectional study was performed to ascertain the HSV-2 prevalence among 919 persons attending an STD clinic in northern Italy. A HSV-2-specific glycoprotein G-2-based immunoglobulin G enzyme-linked immunoabsorbent assay (Gull/Meridian ELISA; Meridian Diagnostics, Cincinnati, OH) was used and validated against Western blot analysis. RESULTS: A prevalence of 24.6% was found without differences between males and females. Seroprevalence increased with age and number of partners during the previous year. Compared with Western blot analysis, the Gull/Meridian ELISA showed a sensitivity of 91.9% and a specificity of 98%, and positive and negative predictive values of 93.9% and 97.4%, respectively. CONCLUSION: This is the first Italian survey of HSV-2 infection conducted with a properly validated, Food and Drug Administration-approved, type specific serologic method in a high-risk population. It is likely that between one to three million adults are infected with HSV-2. PMID- 10821605 TI - Seroprevalence and risk factors of hepatitis B, hepatitis C, and human cytomegalovirus among HIV-infected and high-risk uninfected adolescents: findings of the REACH Study. Adolescent Medicine HIV/AIDS Research Network. AB - BACKGROUND AND OBJECTIVES: In adolescents and young adults, multiple studies have identified sexual activity and behaviors as significant risk factors for acquiring both human cytomegalovirus (HCMV) and hepatitis B virus (HBV). However, there are no reports on the prevalence or risk factors for infection of these viruses and hepatitis C virus (HCV) in an adolescent population with sexually acquired HIV. GOALS: To examine the seroprevalence and risk factors of HBV, HCV, and HCMV infection in a population of HIV-infected male and female adolescents and in an age- and risk behavior-matched HIV-uninfected cohort. STUDY DESIGN: A cross-sectional analysis of HBV, HCV, and HCMV infections in a cohort of HIV infected and HIV-uninfected adolescents. RESULTS: Adolescent males infected with HIV were more likely to have evidence of HBV and HCMV infection than HIV uninfected males (23.7% versus 0%, respectively, for HBV, P = 0.008; 79.7% versus 50%, respectively, for HCMV, P = 0.004). HIV-infected females were more likely to have evidence of HCMV infection (78.5% versus 61.4%, P = 0.003) than HIV uninfected females. No significant difference was found for HBV infection in the two groups of females. The rate of HCV infection (1.6%) was too small to make comparisons between the groups. To determine whether the differences in infection rates for HBV and HCMV could be explained by factors other than HIV status, a variety of possible risk factors were examined using univariate and multivariate analyses. A significant risk factor for HBV and HCMV infections for males was a homosexual or bisexual orientation. For females, a risk factor for HBV infection was having more than 10 lifetime sexual partners; for HCMV infection, HIV infection was the only risk factor. In addition, in the HIV-infected cohort, 15% of females and 36% of males who were seropositive for HBV had evidence of active HBV infection. CONCLUSIONS: These results emphasize the need for continued development of primary and secondary prevention programs and clinical screening and treatment for HBV and HCMV in adolescents. PMID- 10821606 TI - Species delimitation in systematics: inferring diagnostic differences between species. AB - Species are fundamental units in studies of systematics, biodiversity and ecology, but their delimitation has been relatively neglected methodologically. Species are typically circumscribed based on the presence of fixed (intraspecifically invariant or non-overlapping) diagnostic morphological characters which distinguish them from other species. In this paper, we argue that determining whether diagnostic characters are truly fixed with certainty is generally impossible with finite sample sizes and we show that sample sizes of hundreds or thousands of individuals may be necessary to have a reasonable probability of detecting polymorphisms in diagnostic characters at frequencies approaching zero. Instead, we suggest that using a non-zero frequency cut-off may be a more realistic and practical criterion for character-based species delimitation (for example, allowing polymorphisms in the diagnostic characters at frequencies of 5% or less). Given this argument, we then present a simple statistical method to evaluate whether at least one of a set of apparently diagnostic characters is below the frequency cut-off. This method allows testing of the strength of the evidence for species distinctness and is readily applicable to empirical studies. PMID- 10821607 TI - Long-distance colonization and radiation in gekkonid lizards, Tarentola (Reptilia: Gekkonidae), revealed by mitochondrial DNA sequences. AB - Morphological systematics makes it clear that many non-volant animal groups have undergone extensive transmarine dispersal with subsequent radiation in new, often island, areas. However, details of such events are often lacking. Here we use partial DNA sequences derived from the mitochondrial cytochrome b and 12S rRNA genes (up to 684 and 320 bp, respectively) to trace migration and speciation in Tarentola geckos, a primarily North African clade which has invaded many of the warmer islands in the North Atlantic Ocean. There were four main invasions of archipelagos presumably by rafting. (i) The subgenus Neotarentola reached Cuba up to 23 million years (Myr) ago, apparently via the North Equatorial current, a journey of at least 6000 km. (ii) The subgenus Tarentola invaded the eastern Canary Islands relatively recently covering a minimum of 120 km. (iii) The subgenus Makariogecko got to Gran Canaria and the western Canary Islands 7-17.5 Myr ago, either directly from the mainland or via the Selvages or the archipelago of Madeira, an excursion of 200-1200 km. (iv) A single species of Makariogecko from Gomera or Tenerife in the western Canaries made the 1400 km journey to the Cape Verde Islands tip to 7 Myr ago by way of the south-running Canary current. Many journeys have also occurred within archipelagos, a minimum of five taking place in the Canaries and perhaps 16 in the Cape Verde Islands. Occupation of the Cape Verde archipelago first involved an island in the northern group, perhaps Sao Nicolau, with subsequent spread to its close neighbours. The eastern and southern islands were colonized from these northern islands, at least two invasions widely separated in time being involved. While there are just three allopatric species of Makariogecko in the Canaries, the single invader of the Cape Verde Islands radiated into five, most of the islands being inhabited by two of these which differ in size. While size difference may possibly be a product of character displacement in the northern islands, taxa of different sizes reached the southern islands independently. PMID- 10821608 TI - Habitat loss and raptor predation: disentangling long- and short-term causes of red grouse declines. AB - The number of red grouse (Lagopus lagopus scoticus) shot in the UK has declined by 50% during the 20th century This decline has coincided with reductions in the area of suitable habitat and recoveries in the populations of some avian predators. Here we use long-term records of shooting bags and a large-scale manipulation of raptor density to disentangle the effects of habitat loss and raptor predation on grouse populations. The numbers of grouse harvested on the Eskdale half of Langholm Moor in southern Scotland declined significantly during 1913-1990 and grouse bags from the whole moor from 1950 to 1990 exhibited an almost identical but non-significant trend. Hen harriers (Circus cyaneus) and peregrine falcons (Falco peregrinus) were absent or bred at low densities on this moor throughout this period but heather-dominant vegetation declined by 48% between 1948 and 1988. Harrier and peregrine breeding numbers on Langholm Moor increased to high levels following protection in 1990 whilst grouse density and grouse bags declined year after year until shooting was abandoned in 1998. The prediction of a peak in grouse bags on Langholm Moor in 1996 based on the patterns of bags during 1950-1990 was supported by the observed peaks in 1997 on two nearby moors with few raptors which formerly cycled in synchrony with Langholm Moor. This study demonstrates that, whilst long-term declines in grouse bags were most probably due to habitat loss, high levels of raptor predation subsequently limited the grouse population and suppressed a cycle. This study thus offers support to theoretical models which predict that generalist predators may suppress cycles in prey populations. PMID- 10821609 TI - Parasite-associated growth enhancement in a fish-cestode system. AB - Parasites impose an energetic cost upon their hosts, yet, paradoxically instances have been reported in which infection is associated with enhanced, rather than diminished, host growth rates. Field studies of these parasite effects are problematic, since the pre-infection condition of the hosts is generally unknown. Here, we describe a laboratory experiment in which the growth rate and body condition of 76 laboratory-reared three-spined stickleback fishes were examined before, during and after each fish was fed the infective stage of the parasitic cestode Schistocephalus solidus. Twenty-one of these fishes went on to become infected by the cestode. Fishes were individually housed and provided with an abundant food supply to eliminate the potentially masking effects of variable competitive ability. Infection occurred independently of fish gender, size, body condition or pre-exposure growth rate. After exposure to the cestode, infected fishes grew faster (excluding parasite weight) and maintained a similar or better body condition compared with uninfected fishes, despite developing enlarged spleens. The accelerated growth could not be explained by reduced gonadal development. This result, one of few demonstrations of parasite-associated growth enhancement in fishes, is discussed with respect to other such parasite systems. PMID- 10821611 TI - Crickets with extravagant mating songs compensate for predation risk with extra caution. AB - Modern models for the evolution of conspicuous male mating displays assume that males with conspicuous displays must bear the cost of enhanced predation risk. However, if males can compensate behaviourally for their increased conspicuousness by acting more cautiously towards predators, they may be able to lower this cost. In the field cricket Gryllus integer, males call to attract females, and differ in their durations of uninterrupted trilling (calling-bout lengths). Differences among males in calling-bout lengths are heritable, and females prefer males with longer calling bouts. In this study, males with longer, more conspicuous songs behaved more cautiously than males with shorter songs on two different tests of predator avoidance. They took longer to emerge from a safe shelter within a novel, potentially dangerous environment, and they ceased calling for a longer time when their calls were interrupted by a predator cue. Thus, these males appear to compensate behaviourally for their more conspicuous mating displays. Additionally, latencies to emerge from a shelter in the novel environment were consistent over time for both individual males from the field and males that had been reared in the laboratory, indicating that the differences in latency among males may be heritable. PMID- 10821610 TI - Experimentally activated immune defence in female pied flycatchers results in reduced breeding success. AB - Traditional explanations for the negative fitness consequences of parasitism have focused on the direct pathogenic effects of infectious agents. However, because of the high selection pressure by the parasites, immune defences are likely to be costly and trade off with other fitness-related traits, such as reproductive effort. In a field experiment, we immunized breeding female flycatchers with non pathogenic antigens (diphtheria-tetanus vaccine), which excluded the direct negative effects of parasites, in order to test the consequences of activated immune defence on hosts' investment in reproduction and self-maintenance. Immunized females decreased their feeding effort and investment in self maintenance (rectrix regrowth) and had lower reproductive output (fledgling quality and number) than control females injected with saline. Our results reveal the phenotypic cost of immune defence by showing that an activated immune system per se can lower the host's breeding success. This may be caused by an energetic or nutritional trade-off between immune function and physical workload when feeding young or be an adaptive response to 'infection' to avoid physiological disorders such as oxidative stress and immunopathology. PMID- 10821612 TI - The Bateman gradient and the cause of sexual selection in a sex-role-reversed pipefish. AB - As a conspicuous evolutionary mechanism, sexual selection has received much attention from theorists and empiricists. Although the importance of the mating system to sexual selection has long been appreciated, the precise relationship remains obscure. In a classic experimental study based on parentage assessment using visible genetic markers, more than 50 years ago A. J. Bateman proposed that the cause of sexual selection in Drosophila is 'the stronger correlation, in males (relative to females), between number of mates and fertility (number of progeny)'. Half a century later, molecular genetic techniques for assigning parentage now permit mirror-image experimental tests of the 'Bateman gradient' using sex-role-reversed species. Here we show that, in the male-pregnant pipefish Syngnathus typhle, females exhibit a stronger positive association between number of mates and fertility than do males and that this relationship responds in the predicted fashion to changes in the adult sex ratio. These findings give empirical support to the idea that the relationship between mating success and number of progeny, as characterized by the Bateman gradient, is a central feature of the genetic mating system affecting the strength and direction of sexual selection. PMID- 10821613 TI - Adaptive female choice for middle-aged mates in a lekking sandfly. AB - Most theoretical models of age-related mate choice predict that females should prefer older males because they have proven survival ability. An alternative view is that older males represent inferior mates because of negative genetic correlations between early and late fitness components, or because older males have traded off longevity against other fitness components, have accumulated deleterious germ-line mutations, or are less well adapted to current conditions than more recently born individuals. While numerous studies have reported female choice for older males, few have explicitly examined the fitness consequences of such a preference. We present evidence from a lekking sandfly, Lutzomyia longipalpis, showing that choosy females discriminate against older males and gain a fitness benefit from their choice. When permitted free choice from an aggregation consisting of males aged zero to two days (young), four to six days (middle-aged) and eight to ten days (old), females preferentially mated with middle-aged males, but all measures of female reproductive success were independent of male age. In contrast, when a second set of females was randomly assigned single virgin males of known age, the eggs of those paired to old mates exhibited lower hatching success than the eggs of females mated to young or middle-aged males. These results suggest that females avoid mating with older males because they represent poorer quality mates. Age-related differences in male quality may have a genetic basis, but could equally well arise through a phenotypic decline in sperm quality or sperm transfer ability with male age. The lack of evidence of female discrimination against older males from other studies may be because these did not explore the reproductive success of the full age range of males. PMID- 10821614 TI - The consequences of inbreeding for recognizing competitors. AB - Extreme inbreeding will compromise an animal's ability to discriminate between individuals and, thus, assess familiarity and kinship with conspecifics. In rodents, a large component of individual recognition is mediated through chemical communication. The counter-marking of competitor males' scent marks provides a measure of discrimination between their own scent and that from other individuals. We investigated whether males in common outbred (ICR(CD-1) and TO) and inbred (BALB/c) strains of laboratory mice could recognize the urinary scents of other individuals by measuring their investigation and counter-marking responses. Dominant males of outbred strains investigated and counter-marked scents from other males, whether of the same or another strain. Dominant inbred BALB/c males investigated but did not counter-mark their own strain scents, counter-marking only those from another strain. They did not use environmentally induced status differences in odours to recognize scents from other males. The inability of the inbred mice to discriminate between their own scent marks and those of other males is likely to alter their competitive behaviour, which could influence responses in experiments and the welfare of caged laboratory mice. PMID- 10821615 TI - Kin recognition and the 'armpit effect': evidence of self-referent phenotype matching. AB - In species with multiple paternity or maternity, animals may best assess their relatedness to unfamiliar conspecifics by comparing their own phenotype(s) with those of unidentified individuals. Yet whether animals can recognize kin through self-matching is controversial. Because golden hamsters (Mesocricetus auratus) mate multiply and can produce multiply sired litters, they were tested for their ability to use self-matching for kin recognition. Hamsters that were reared only with non-kin since birth responded differentially to odours of unfamiliar relatives and non-relatives. Postnatal association with kin was not necessary for this discrimination. Prenatal learning was unlikely because of delayed production and perception of social odours. To our knowledge, this is the first demonstration that a vertebrate can use its own phenotype for kin-recognition purposes without prior experience with kin. By using itself as a referent, rather than its siblings or parents, a golden hamster may be better able to direct nepotism towards the most appropriate individuals. Kin discrimination via self inspection may be especially important in nepotistic contexts (to identify most closely related conspecifics), whereas inclusion of the phenotypes of close kin as referents may be favoured in mate-choice contexts (to identify all related individuals). PMID- 10821616 TI - Individual reproductive success and effective population size in the greater white-toothed shrew Crocidura russula. AB - In order to investigate the determinants of effective population size in the socially monogamous Crocidura russula, the reproductive output of 44 individuals was estimated through genetic assignment methods. The individual variance in breeding success turned out to be surprisingly high, mostly because the males were markedly less monogamous than expected from previous behavioural data. Males paired simultaneously with up to four females and polygynous males had significantly more offspring than monogamous ones. The variance in female reproductive success also exceeded that of a Poisson distribution (though to a lesser extent), partly because females paired with multiply mated males weaned significantly more offspring. Polyandry also occurred occasionally, but only sequentially (i.e. without multiple paternity of litters). Estimates of the effective to census size ratio were ca. 0.60, which excluded the mating system as a potential explanation for the high genetic variance found in this shrew's populations. Our data suggest that gene flow from the neighbourhood (up to one third of the total recruitment) is the most likely cause of the high levels of genetic diversity observed in this shrew's subpopulations. PMID- 10821617 TI - Perception of female reproductive state from vocal cues in a mammal species. AB - While acoustic signalling by males is known to affect male-male competition, mate attraction and the timing of ovulation, the extent to which sexual selection has shaped the evolution of female acoustic signals is poorly understood. Among mammals, experimental evidence indicates that females attract mating partners by using olfactory and visual signals to advertise their reproductive state. Whether or not males ascertain female reproductive state from vocal signals has, however, never been systematically tested. In this study, we use playbacks of recorded vocalizations to demonstrate that male Barbary macaques, Macaca sylvanus, can discriminate between female copulation calls given at different stages of the oestrous cycle, responding more strongly to those given around the time when conception is most likely to occur. Acoustic analysis suggests that the mean dominant frequency of call units and a number of temporal parameters could provide males with the information necessary to discern the proximity of ovulation in this way Our results provide the first experimental evidence that the calls of female mammals may contain information on reproductive state, which males can perceive and use in such a way as to increase their reproductive success. PMID- 10821618 TI - Interspecies semantic communication in two forest primates. AB - West African Diana monkeys (Cercopithecus diana) and Campbell's monkeys (Cercopithecus campbelli) frequently form mixed-species associations. Males of both species produce acoustically distinct alarm calls to crowned eagles (Stephanoaetus coronalus) and leopards (Panthera pardus), two of their main predators. Field playback experiments were conducted to investigate whether Diana monkeys respond to Campbell's alarm calls and whether they understand the calls' semantic content. Diana monkeys responded to playback of Campbell's leopard or eagle alarm calls as though the original predator were present. In a second experiment, Diana monkeys were primed with either Campbell's eagle or leopard alarm calls and then subsequently probed with the vocalizations of a crowned eagle or a leopard. Results showed that monkeys used the semantic information conveyed by the Campbell's alarm calls to predict the presence of a predator. The data are consistent with the hypothesis that non-human primates are able to use acoustic signals of diverse origin as labels for underlying mental representations. PMID- 10821619 TI - Dishonest signalling in a fiddler crab. AB - Animal communication theory predicts that low-frequency cheating should be common in generally honest signalling systems. However, perhaps because cheats are designed to go undetected, there are few examples of dishonest signals in natural populations. Here we present what we believe is the first example of a dishonest signal which is used commonly by males to attract mates and fight sexual rivals. After losing their large claw male fiddler crabs (Uca annulipes) grow a new one which has less mass, is a less effective weapon and costs less to use in signalling than an equivalent-length claw of the original form. Males with original claws do not differentially fight males with regenerated claws even though they are likely to win. Regenerated claws effectively bluff fighting ability and deter potential opponents before they fight. During mate searching, females do not discriminate against males with low-mass, regenerated claws, indicating that they are deceived as to the true costs males pay to produce sexual signals. Up to 44% of males in natural populations have regenerated claws, a level unanticipated by current signalling theory. The apparent rarity of cheating may be an artefact of the usual difficulty of detecting cheats and dishonesty may be quite common. PMID- 10821620 TI - Testing Mullerian mimicry: an experiment with wild birds. AB - Experiments with wild birds feeding on pastry 'prey' were performed to test competing theories of Mullerian mimicry Conventional theories predict that all resemblances between defended prey will be mutually advantageous and, hence, Mullerian. In contrast, unconventional theories predict that, if there are inequalities in defences between mimetic species, the less well-defended prey may dilute the protection of the better defended species in a quasi-Batesian manner. This unconventional prediction follows from an assumption that birds learn about the edibilities of prey using rules of Pavlovian learning. We report on two experiments, each lasting 40 days, which showed that a moderately defended prey can dilute the protection of a better defended mimic in a quasi-Batesian fashion, but can add protection to a mimic which has the same moderate levels of defence. These results match predictions of unconventional theories of mimicry and go some way to resolving the long-running arguments over the nature of Mullerian mimicry. PMID- 10821621 TI - Evidence for a rule governing the avoidance of superfluous escape flights. AB - When an imminent attack by a predator on a group of birds is signalled to non detectors only by the departure of the detector, non-detectors may make time wasting false-alarm flights in response to mistaken or non-predator-driven departures. The frequency of false-alarm flights might be reduced if group members assess the reason for single departures before responding. Immediate flights should only occur after multiple simultaneous departures, because these are only likely to be generated by an attack. The response delay between the detectors' departure and the next birds that respond should then be dependent on the number of detectors. On sparrowhawk attack, response delays in redshanks decreased significantly as detector number increased, controlling for raptor conspicuousness and proximity, and flock size and spacing. If response delay is modified because of risk dilution, it should increase with flock size and, consequently, the rate of alarm flights due to mistakes should decrease. However, response delay did not increase and flight frequency due to misidentification of non-raptors or non-predator-driven departures did not decrease with flock size. Significantly more feeding time was lost by birds in small flocks, suggesting that the dilution effect decreased the cost of each false-alarm flight rather than their frequency. PMID- 10821622 TI - Subject and interview characteristics affecting reliability of the Diagnostic Interview Schedule for Children. AB - Despite the well-known difficulties in obtaining reliable and valid assessments of child psychopathology, investigators generally have not examined the influence of factors such as subject characteristics or the specific assessment procedures themselves on the validity of the information obtained. To address these issues, this special section presents four studies of the Diagnostic Interview Schedule for Children, in which investigators examined the impact of a range of variables on the reliability of its symptom and diagnostic information. Factors studied include interview structural characteristics; question length, complexity, and placement within the interview; and interview subject characteristics. Overall findings suggest that interview and subject characteristics exert important influences on the data obtained, and that novel approaches, such as allowing subjects a greater role in the ordering of questions to be answered, may improve the precision and accuracy of such measures of children's psychopathology. PMID- 10821623 TI - Informant-based determinants of symptom attenuation in structured child psychiatric interviews. AB - Informant-related determinants of item attenuation, that is, the drop-off in symptom endorsement rates at retest, were examined in an enriched community subsample of 245 parent-child pairs drawn from the National Institute of Mental Health Methods for Epidemiology of Child and Adolescent Mental Disorders Study. Youngsters and their parents were interviewed with the Diagnostic Interview Schedule for Children (Version 2.3; DISC-2.3) on two occasions with a mean test retest interval of 12 days. Item attenuation rates were high for both informants, with adults failing to confirm 42% and children 58% of baseline responses at retest. Stepwise regressions revealed that item attenuation at DISC-P retest was higher for adult informants who were younger, and who reported on older and less impaired children. On the DISC-C, attenuation was higher for children who were less impaired, rated as doing worse in school, and who had a longer test-retest interval. These results are broadly consistent with past studies examining the determinants of attenuation and test-retest reliability and have implications for the design and use of structured diagnostic instruments. PMID- 10821624 TI - Features of interviews questions associated with attenuation of symptom reports. AB - Previous studies have suggested that discrepant reporting in a test-retest reliability paradigm is not purely random measurement error, but partly a function of a systematic tendency to say "no" during retest to questions answered positively at initial testing ("attenuation"). To examine features of interview questions that may be associated with attenuation, three raters independently assessed the structural and content features of questions from the Diagnostic Interview Schedule for Children (version 2.3) and linked these to data from a test-retest reliability study of 223 community respondents (parent and child reports). Results indicated that for both parent and youth reports, item features most strongly associated with attenuation were (a) being a "stem" question (asked of all respondents, regardless of any skip structure); (b) question placement in the first half of the interview; (c) question length; (d) question complexity; or (e) requiring assessment of the timing, duration, or frequency of a symptom. Findings may be explained by participants' conscious efforts to avoid further questions or by their learning more about the nature and purpose of the interview as they gain more experience; alternatively, findings may represent a methodological artifact of structured interview design. PMID- 10821625 TI - Who's up first? Testing for order effects in structured interviews using a counterbalanced experimental design. AB - A growing body of research suggests that, apart from the wording of specific questions, various aspects of the interview process itself may affect the reliability of information provided by research participants. To examine whether the order of presentation of specific diagnostic modules affects the likelihood of subjects' yes/no responses within the Diagnostic Interview Schedule for Children (DISC), the authors used a counterbalanced design, presenting two DISC diagnostic modules to children and their parents in standard or reversed order. Results indicate that the order of module administration exerts effects on the total numbers of symptoms endorsed, level of impairment, and the likelihood of meeting diagnostic criteria, regardless of whether the information is provided by parent or child respondents. Future child and adult assessment measures should take these difficulties fully into account through novel approaches to instrument design and interview procedures. PMID- 10821626 TI - Interviewing as communication: an alternative way of administering the Diagnostic Interview Schedule for Children. AB - Parents of 24 children referred to an outpatient psychology clinic (mean child age 10.8, range 6-15) were administered the Diagnostic Interview Schedule for Children Version 2.3 (DSIC-2.3) twice in a 1-week test-retest reliability design (mean retesting interval = 7.5 days, range = 6-11 days). An alternative mode of administration of the DISC, based on communication principles, was used, involving (a) a schematic representation of the areas to be covered; (b) definition of a common language for the categories, diagnoses, and criteria; and (c) the respondent being allowed to select the order in which the diagnostic areas were covered. The DISC items and modules were unchanged. Symptom scores derived from the DISC were highly reliable over 1 week (average ICC = .85, range = .67-.95) and showed no attenuation from Time 1 to Time 2. Reliability of DSM diagnoses averaged kappa = .80 (range = .63-1.0). There was no significant attenuation in diagnoses from Time 1 to Time 2. Overall, this alternative way of administering the DISC appears to have promise for reducing attenuation and boosting the reliability-and ultimately the validity--of child psychiatric diagnoses. Further investigations of the mechanisms underlying these effects, and further studies with child and adolescent respondents and nonreferred community samples are recommended. PMID- 10821627 TI - Prevalence of attention-deficit/hyperactivity disorder symptoms in 4- to 17-year old children in the general population. AB - The purposes of this study were (a) to estimate the prevalence of Attention Deficit/Hyperactivity Disorder (AD/HD) symptoms in the general preschool and school population; and (b) to analyze the influence of gender, age, and socioeconomic status (SES) variables on AD/HD symptoms. Out of the 80,000 preschool and schoolchildren living in Manizales, Colombia, a random sample of 540 children was selected. Two gender, three age (4- to 5-year olds, 6- to 11 year olds, and 12- to 17-year olds), and three SES (low, middle, and high) groups were used. The 18 DSM-IV symptoms corresponding to AD/HD Criterion A were assessed on a scale of 0 (never) to 3 (almost always). All three demographic variables established statistically significant differences: AD/HD symptoms were more frequent in 6- to 11-year-old, low-SES, male participants. DSM-IV Criterion A for AD/HD was fulfilled by 19.8% of the boys and 12.3% of the girls. However, this difference was marginally significant only in the AD/HD Subtype I: Combined. It was concluded that demographic variables are significant correlates of the AD/HD diagnosis. The prevalence found in this study was higher than usually reported, even though only the symptomatic DSM-IV AD/HD criterion was analyzed. We failed to confirm the assumed AD/HD gender ratio. PMID- 10821628 TI - Utility of behavior ratings by examiners during assessments of preschool children with attention-deficit/hyperactivity disorder. AB - This study examines the clinical utility of behavior ratings made by nonclinician examiners during assessments of preschool children with Attention Deficit/Hyperactivity Disorder (AD/HD). Matched samples of children with (n = 127) and without (n = 125) AD/HD were utilized to test the internal, convergent, concurrent, and incremental validity of ratings completed by examiners on the Hillside Behavior Rating Scale (HBRS). Results indicated that HBRS ratings were internally consistent, possessed sufficient interrater reliability, and were significantly associated with parent and teacher reports of AD/HD when controlling for age, gender, intelligence, and symptoms of other psychopathology. HBRS ratings also were significantly associated with other measures of functioning, and provided a significant increment in the prediction of impairment over parent and teacher report alone. These findings suggest that behavioral ratings during testing provide a unique source of clinical information that may be useful as a supplement to parent and teacher reports. PMID- 10821630 TI - DmGluRA, a Drosophila metabotropic glutamate receptor, activates G-protein inwardly rectifying potassium channels in Xenopus oocytes. AB - Xenopus oocytes were coinjected with cDNAs encoding the Drosophila melanogaster metabotropic glutamate receptor (DmGluRA) and two mammalian G-protein inwardly rectifying potassium channel subunits (GIRK1 and GIRK2). Glutamate and two vertebrate group II mGluR agonists (order of potency: LY 354740 > glutamate > DCG IV) elicited inwardly rectifying potassium currents. These inward currents were sensitive to cesium and barium. They were also blocked by two group II specific antagonists MCCG and APICA (IC50s 97.5 and 200 microM, respectively) and not affected by a group I antagonist (AIDA). Finally, the A-protomer of PTX reduced the glutamate-induced GIRK currents. This study is the first characterization of an invertebrate mGluR-mediated GIRK currents via a PTX-sensitive G protein. PMID- 10821629 TI - Relations of age to cognitive and motivational elements of impulse control in boys with and without externalizing behavior problems. AB - In a cross-sectional study of 83 unmedicated boys, 6 to 16 years of age (M = 10.6, SD = 2.1), attending public (N = 48) and therapeutic schools for behaviorally disturbed children (N = 35), we examined relations of externalizing psychopathology to age-dependent change in performance on cognitive and motivational dimensions of impulse control assessed by laboratory tasks. When we controlled for internalizing symptoms and IQ or school achievement, all children showed improving competence with increasing age on both dimensions over the age range of the sample. Children with externalizing problems performed more poorly on both dimensions at all ages than children without such problems. Comparing age dependent competence for the two groups, a model of convergent maturation in cognitive aspects of impulse control, and a model depicting a stable deficit in motivational aspects of impulse control in those children with externalizing behavior problems, relative to those without such problems, emerged. Studies of individual growth in impulse control, together with correlates of growth, are needed to validate these observations. PMID- 10821631 TI - Immunohistochemical investigation of the vomeronasal organ. Nitric oxide synthase expression in the mouse during postnatal development. AB - The expression of nitric oxide synthase type I (NOS-I), the key enzyme for the synthesis of the gaseous neurotransmitter nitric oxide, was investigated by means of immunohistochemistry in the vomeronasal organ (VNO) of mice from postnatal day 1 for 2 months. The results show that NOS is expressed in extrinsic nerve supplying the developing erectile tissue of VNO (the so-called VNO pump) as well as blood vessels in the connective tissue laying under the receptor epithelium at postnatal day 1. At 8, 15 and 21 postnatal days, and at 2 months the density of NOS-1-immunoreactive nerves goes along with the development of the erectile tissue. From postnatal day 8 onwards, NOS-1-immunoreactive fibers are found also in the vicinity of the VNO glands. These data suggest that nitric oxide (NO) modulates VNO activity early after birth in the mouse. PMID- 10821632 TI - Progesterone microinjections into the pontine reticular formation modify sleep in male and female rats. AB - It has been reported that progesterone (P4) induces changes in sleep, but the brain regions involved in these actions are unknown. We studied the effects of P4 microinjections into the pontine reticular formation (PRF) upon rat sleep. Intact adult male and ovariectomized female rats were unilaterally injected with P4 into the PRF and the sleep-waking cycle was recorded for 6 h. P4 (1.0 and 5.0 microg/0.2 microl) did not modify sleep, but at a higher dose (7.5 microg/0.2 microl) it produced a marked decrease in rapid eye movement sleep (REM) latency in both male (55%) and female (63%) rats. A non-significant increase in the number of REM episodes was observed after P4 administration. These findings suggest that P4 should participate in the mechanisms related to REM initiation in the rat through its effects in the PRF. PMID- 10821633 TI - Epitope mapping of LB509, a monoclonal antibody directed against human alpha synuclein. AB - Alpha-synuclein is a 140 amino acid protein that forms the major component of the abnormal filaments that make up the Lewy bodies and Lewy neurites of Parkinson's disease and dementia with Lewy bodies. It is also the major component of the filamentous glial cytoplasmic inclusions of multiple system atrophy. Here we have used recombinant alpha-synucleins and peptide competition to show that the monoclonal anti-alpha-synuclein antibody LB509 recognizes amino acids 115-122 of human alpha-synuclein. The antibody strongly labelled filaments extracted from multiple system atrophy brain, showing the presence of residues 115-122 of alpha synuclein. LB509 failed to react with mouse, rat and zebra finch alpha-synuclein, because of amino acid differences with human alpha-synuclein. Since LB509 recognizes human but not rodent alpha-synuclein, it will be a useful reagent for the characterization of mouse lines transgenic for human alpha-synuclein. PMID- 10821634 TI - Effect of trilinolein on the activity and gene expression of superoxide dismutase in cultured rat brain astrocytes. AB - Cerebrovascular disease is one of the major causes of morbidity and mortality in recent. Oxygen free radicals produced during cerebral infarction increases the damage to neurons. Superoxide dismutase (SOD) is the endogenous antioxidant enzyme that can effectively scavenge superoxide radicals. Trilinolein is a lipophilic antioxidant purified from the herb of Panax pseudoginseng. In the cultured rat brain astrocytes (RBA), the activity of SOD (both Cu,Zn-SOD and Mn SOD subtypes) was markedly increased by incubation with trilinolein at low concentration (0.1 microM) for 2 days. This stimulatory effect of trilinolein was not related to the incubating concentration. However, long-term (7 days) incubation with trilinolein at same concentration decreased the activity. Similar changes were also observed in the gene expression of SOD in RBA; short-term (2 days) incubation of RBA by 0.1 microM trilinolein increased the mRNA level that was lowered in RBA received a long-term incubation with 0.1 microM trilinolein. This result shows that trilinolein is an effective antioxidant to increase the activity of SOD in RBA which would be beneficial to neurons subjected to oxygen free radical damage. However, long-term medication of antioxidant shall be concerned. PMID- 10821635 TI - Modulation of agonist stimulated adenylyl cyclase and GTPase activity by L-pro-L leu-glycinamide and its peptidomimetic analogue in rat striatal membranes. AB - L-prolyl-L-leucyl-glycinamide (PLG), also known as melanocyte-stimulating hormone release inhibiting factor (MIF-1), is an endogenous brain tripeptide. Previous studies have shown that PLG, and its peptidomimetic analogues, render dopamine D2 receptors more responsive to agonists by maintaining the high-affinity binding state of the receptors. In the present study, we examined the effect PLG and its analogue 3(R)-[(2(S)-pyrrolidylcarbonyl)amino]-2-oxo-1-pyrrolidineacetam ide (PAOPA) on dopamine-stimulated adenylyl cyclase and NPA-stimulated GTPase activity in rat striatal membranes. Dopamine-stimulated adenylyl cyclase activity was inhibited by both PLG and PAOPA in a dose-dependent manner, whereas R(-) propylnorapomorphine (NPA)-stimulated low Km GTPase activity was significantly increased by 1 microM PLG or 1 nM PAOPA. These results suggest that PLG and PAOPA maintain the high affinity state of the D2 receptor by increasing GTP hydrolysis through stimulation of agonist-induced GTPase activity. PMID- 10821636 TI - Lidocaine produces a shunt in rat [correction of rats] thalamocortical neurons, unaffected by GABA(A) receptor blockade. AB - Low concentrations of lidocaine reversibly decrease input resistance and shunt action potentials in neurons of the ventral posterolateral thalamic nucleus (VPL) in vitro. Using differential interference contrast infrared videomicroscopy and whole-cell patch-clamp techniques in rat brain slices, we studied the effects of bicuculline methobromide to test whether the lidocaine-induced shunt in VPL neurons is mediated by GABA(A) receptors. Bicuculline (50 microM) restored tonic firing rates to control values following reduction by lidocaine (10 microM). However, bicuculline did not influence lidocaine's actions to decrease slope resistance over the voltage range from approximately -90 mV to spike threshold. Likewise, bicuculline did not affect the shunt-induced reductions of spike afterhyperpolarizations produced by lidocaine. The results of this study imply that the effects of low lidocaine concentrations in thalamocortical neurons are not mediated by GABA(A) receptors. PMID- 10821637 TI - Immediate increase in benzodiazepine binding in rat brain after a single brief experience in the plus maze: a paradoxical effect. AB - A single drug-free experience in the elevated plus-maze is well documented to reduce the behavioral effects of benzodiazepines (BZs) in subsequent tests. To ascertain the possible role of altered BZ receptor binding to in this phenomenon, rats received a 5-min exposure to the elevated plus maze and were immediately sacrificed. Receptor autoradiography revealed that [3H]flunitrazepam binding was significantly elevated in several amygdaloid and hippocampal nuclei (range: 13 23%); [3H]muscimol binding in adjacent sections was not significantly altered. These results suggest that BZ receptors can change very rapidly in response to anxiogenic conditions. However, the unexpected finding that [3H]flunitrazepam binding is increased by maze exposure suggests that the subsequent loss of BZ anxiolytic effects in the plus-maze is probably unrelated to alterations in BZ binding in brain. PMID- 10821638 TI - Corticostriatal projections from distal and proximal forelimb representations of the monkey primary motor cortex. AB - Corticostriatal projections from one distal and two proximal subregions in the forelimb representation of the primary motor cortex (MI) were examined in the macaque monkey. The distal and proximal subregions in the anterior bank of the central sulcus (distal and proximal-bank subregions) and the proximal subregion in the surface of the precentral gyrus (proximal-surface subregion) of the MI were identified using intracortical microstimulation. Different anterograde tracers were then injected into two of these three forelimb subregions of the MI. In the ipsilateral putamen, the distribution areas of corticostriatal fibers from the distal, proximal-bank and proximal-surface subregions were arranged from ventrolateral to dorsomedial in this order. These corticostriatal input zones were largely segregated from one another. PMID- 10821639 TI - Tyrosine hydroxylase and aromatic L-amino acid decarboxylase do not coexist in neurons in the human anterior cingulate cortex. AB - Immunoreactivity for aromatic L-amino acid decarboxylase (AADC), the second step dopamine-synthesizing enzyme, was found immunohistochemically in neurons of the human anterior cingulate cortex (ACC). Most of these neurons were located in layers V and VI and subcortical white matter; a small number were occasionally found in layer III. Double immunohistochemistry for tyrosine hydroxylase (TH: the first step dopamine-synthesizing enzyme) and AADC revealed that no neuronal cell bodies in the ACC were doubly immunostained for TH and AADC, suggesting that these TH-only- or AADC-only-immunoreactive neurons were not dopaminergic. AADC neurons in the human ACC might transform L-DOPA to dopamine, droxidopa to noradrenaline, and/or 5-hydroxytryptophan to serotonin. PMID- 10821640 TI - RET-RGS, a retina-specific regulator of G-protein signaling, is located in synaptic regions of the rat retina. AB - RGS (regulators of G protein signaling) proteins negatively regulate the alpha subunit of G proteins by accelerating their intrinsic GTPase activity. In a previous work, we reported the cloning of a cDNA encoding for a new RGS protein, RET-RGS. We showed that it is specifically expressed in the retina, notably by photoreceptor cells and that it has an in vitro GAP activity on transducin. To understand the role of RET-RGS, and in particular to determine whether it regulates the phototransduction cascade in photoreceptor cells, RET-RGS was immunolocalized on rat retina sections. Whereas no labeling was detected in outer nor inner segments of photoreceptors cells, dense immunoreactive products were localized in the outer and inner plexiform layers which correspond to the regions of synaptic interplay between the different neurons of the retina including the photoreceptor cells. These results rule out a role of RET-RGS on the phototransduction cascade and suggest that it may participate in retina specific synaptic transductions. PMID- 10821641 TI - Odorant suppression of delayed rectifier potassium current in newt olfactory receptor cells. AB - Effects of odorants on a delayed rectifier potassium current (IK) in newt olfactory receptor cells (ORCs) were investigated using the whole-cell version of the patch-clamp technique. Under voltage clamp, odorants (amyl acetate, limonene and acetophenone) reversibly suppressed I(K) without shifting its I-V curve. An amyl acetate puff completely suppressed I(K) induced by the first step pulse of repetitive depolarizations, suggesting that binding of an odorant molecule to the open channel is not required to block the channel. Although it is known that odorants suppress Na+ and Ca2+ currents (I(Na), I(Ca)) by shifting their inactivation curves to a negative voltage, odorants did not shift the inactivation curve of I(K) significantly. This suggests that odorants suppress I(K) without changing its voltage dependence. Therefore, the blocking mechanisms by odorants of I(K) in ORCs are different from those of I(Na) and I(Ca). PMID- 10821642 TI - Characterization of voltage dependent calcium channels on the lateral olivocochlear efferent fibers of the guinea pig. AB - Using in vitro superfusion technique the release of [3H]-dopamine from the lateral olivocochlear efferent fibers of the cochlea was investigated. Our previous study gave the first neurochemical evidence for the transmitter role of dopamine and proved its neuronal origin. Using specific antagonists now we characterized the voltage-dependent calcium channels (VDCCs) involved in the release of dopamine evoked by electrical stimulation of the cochlear tissue. Verapamil or nifedipine, and Ni2+ failed to affect the release, indicating that neither L-, nor T-type VDCCs are essential for the release process. The fact that omega-conotoxin inhibited the release of dopamine from lateral olivocochlear efferent fibers suggests, that N-type VDCCs are required for the calcium influx during electrical stimulation. These VDCCs could be presynaptic targets of modulation of the dopamine release under pathological conditions or in therapy. PMID- 10821643 TI - Alterations of 3-nitrotyrosine concentration in the cerebrospinal fluid during aging and in patients with Alzheimer's disease. AB - To investigate the significance of nitric oxide (NO)-mediated neuron death in aging and Alzheimer's disease (AD), the 3-nitrotyrosine concentration in the cerebrospinal fluid (CSF) was investigated in neurologically normal controls and patients with AD. The 3-nitrotyrosine concentration and the 3 nitrotyrosine/tyrosine ratio significantly increased with advancing age, whereas the tyrosine concentration was unaltered. In patients with AD, the 3 nitrotyrosine concentration and the 3-nitrotyrosine/tyrosine ratio increased significantly (>six-fold) compared with controls of similar age, and increased significantly with decreasing cognitive functions, whereas the tyrosine concentration did not change. These findings suggest that an activation of tyrosine nitration, increase in nitrated tyrosine-containing proteins, and/or its degradation may be involved in brain aging and play an important role in the pathogenesis of AD. PMID- 10821644 TI - Expression of human GFR alpha-1 (GDNF receptor) at the neuromuscular junction and myelinated nerves. AB - Motor neurons have been known to require a wide variety of neurotrophic factors for their survival. As one of the target-derived trophic factors, glial cell line derived neurotrophic factor (GDNF) has been shown to exert its effects on motor neurons via a receptor complex including GDNF receptor alpha 1 (GFR alpha-1). Immunoreactivity of GFR alpha-1 was observed at myelinated peripheral nerves and neuromuscular junction (NMJ) of human skeletal muscles. Reverse transcriptase polymerase chain reaction (RT-PCR) analyses showed that mRNA of GFR alpha-1 existed in the ventral horn of human spinal cord, but not in the skeletal muscles. The results suggested that GFR alpha-1 might play a key role for uptake and internalization of GDNF at the human NMJ. PMID- 10821645 TI - Macrocyclic chelators with paramagnetic cations are internalized into mammalian cells via a HIV-tat derived membrane translocation peptide. AB - A major obstacle to using paramagnetic MR contrast agents for in vivo cell tracking or molecular sensing is their generally low cellular uptake. In this study, we show that a paramagnetically labeled DOTA chelator derivatized with a 13-mer HIV-tat peptide is efficiently internalized into mammalian cells. Intracellular concentrations were attained that were readily detectable by MR imaging using both gadolinium and dysprosium chelates. Using this paradigm, it should be feasible to internalize a variety of chemically different agents into mammalian cells. PMID- 10821646 TI - O-Alkyl dioleoylphosphatidylcholinium compounds: the effect of varying alkyl chain length on their physical properties and in vitro DNA transfection activity. AB - 1,2-Dioleoyl-sn-3-ethylphosphocholine (EDOPC) has been previously shown be a highly effective DNA transfection reagent in vitro. To assess the effect of alkyl chain length on transfection efficiency, the O-methyl, O-propyl, O-hexyl, O decyl, and O-octadecyl derivatives have been prepared from dioleoylphosphatidylcholine using the corresponding alkyl trifluoromethylsulfonate. The methyl, ethyl, and propyl derivatives formed liposomes which were very large and unilamellar. The ethyl and propyl derivatives were equally efficient at mediating transfection (even in the presence of serum) of BHK cells, but the chemically labile methyl derivative was a much weaker transfection agent. The O-decyl and O-octadecyl compounds, which assume the inverted hexagonal phase in excess water (as determined by X-ray diffraction), were almost inactive after manual agitation in both water and in saline; however, after sonication, these compounds exhibited good transfection activity. The O hexyl derivative displayed novel behavior, assuming the lamellar phase at low and a cubic phase at high ionic strength. All compounds, whether lamellar or not, formed lamellar structures when complexed with DNA. In water, where the hexyl compound dispersed well, sonication diminished transfection activity, whereas at physiological ionic strength, which led to poor manual dispersion, sonication was essential for good transfection. These results emphasize the importance of optimal dispersion of a cationic lipid: too little, and interaction with DNA is handicapped, too much, and the resultant particle transfects poorly. Lipid dispersibility is thus an important variable in assessing lipid transfection agents, and caution is advised in attributing too much significance to chemical structure until interaction with DNA has been optimized. PMID- 10821647 TI - Synthesis of cholesteryl polyamine carbamates: pK(a) studies and condensation of calf thymus DNA. AB - Novel polyamine carbamates have been designed and prepared from cholesterol. Our synthesis uses an orthogonal protection strategy based upon trifluoroacetyl and Boc-protecting groups. These unsymmetrical polyamine carbamates have been prepared from symmetrical (e.g., spermine and thermine) polyamines. Detailed interpretations of (1)H and (13)C NMR spectroscopic data led to the unambiguous assignment of these polyamine carbamates. These target conjugates contain a variety of positive charges distributed along methylene chains. Their pK(a)s have been determined potentiometrically for conjugates substituted with up to five amino functional groups. Condensation of calf thymus DNA into particles was monitored using light scattering at 320 nm. Salt-dependent binding affinity for calf thymus DNA was determined using an ethidium bromide fluorescence quenching assay. These cholesteryl polyamine carbamates are models for lipoplex formation with respect to gene delivery (lipofection), a key first step in gene therapy. PMID- 10821648 TI - Biodistribution and radioimmunotherapy of human breast cancer xenografts with radiometal-labeled DOTA conjugated anti-HER2/neu antibody 4D5. AB - HER2/neu oncogene encodes a 185 kDa trans-membrane protein which is overexpressed in 20-30% of breast and ovarian cancers and portends a poor prognosis. We have studied the targeting and therapy of this oncoprotein with 4D5, a murine monoclonal antibody which recognizes a distinct epitope on the extracelluar domain of HER2/neu. We conjugated the antibody with an active ester of the macrocyclic chelating agent DOTA, radiolabeled the conjugate with either (111)In or (90)Y, and studied the antibody distribution and therapy, respectively, in athymic mice bearing xenografts of MCF7/HER2/neu, a human breast cancer cell line transfected with the HER2/neu oncogene. For the biodistribution of (111)In labeled DOTA-4D5, a high specificity of tumor localization (30% ID/g) was seen with a tumor-to-blood ratio of greater than 2 at 48 h postinjection. Compared to a previously published study with (125)I-labeled 4D5 in beige nude mice bearing NIH3T3/HER2/neu xenografts [De Santes et al. (1992) Cancer Res. 52, 1916-1923], (111)In-labeled 4D5 antibody gave superior antibody uptake in tumor (30% ID/g vs 17% ID/g at 48h). In the therapy study, treatment of the nude mice bearing MCF7/HER2/neu xenografts with 100 microCi (3 microg) of (90)Y-labeled DOTA-4D5 caused a 3-fold reduction of tumor growth compared to untreated controls (injected with human serum albumin) in 40 days. Treatment of animals with 100 microCi of nonspecific antibody (90)Y-labeled DOTA-Leu16 (3 microg) had no tumor growth inhibition. Treatment with unlabeled DOTA-4D5 (3 microg) had a slight effect on tumor growth compared to untreated controls. When analyzed at the level of single animals, no effect was seen in seven of nine animals; however, in two of the animals, tumor growth inhibition was observed. Although a cold antibody therapeutic effect was unexpected at this dose level (3 microg), it may be possible that in some animals that 3 microg of antibody of (90)Y-labeled DOTA-4D5 augmented tumor growth reduction. To further explore the effects of cold antibody treatment alone, animals were treated with 100 or 400 microg of unlabeled 4D5 administered in two doses. These animals showed a 1.7-1.8-fold reduction in tumor growth over 28 days, a result less than that obtained with RIT only. PMID- 10821649 TI - Molecular characterization of a tetramolecular complex between dsDNA and a DNA binding leucine zipper peptide dimer by mass spectrometry. AB - The characterization of sequence-specific noncovalent complexes of the GCN4 peptides and dsDNA using mass spectrometry is reported. The GCN4 peptides belong to a class of proteins which bind to sequence-specific dsDNA and are important in the regulation of gene transcription in yeast. These proteins contain a bZIP structural motif which consists of a basic DNA-binding domain and a leucine zipper dimerization domain. The protein dimers specifically bind double-stranded DNA containing the binding element 5'-ATGA(C/G)TCAT-3' to form a tetramolecular noncovalent complex. Using electrospray ionization, we report the detection of such a specific tetramolecular complex using mass spectrometry. Under conditions necessary for observation of the tetramolecular complex, no ions were detected for the GCN4 peptide dimer or the GCN4 monomer with dsDNA. These observations indicate that the specific interaction of the dsDNA with the protein dimer stabilizes the biologically significant noncovalent complex in the gas phase. Complexes were observed for various lengths of both blunt-ended and cohesive ended double-stranded DNA containing the specific recognition sequence. The binding specificity of the complex was verified with the use of control DNA not containing the recognition sequence and control peptides not known to bind DNA specifically. Additionally, combining limited proteolysis of GCN4 peptide-DNA complexes with mass spectrometric determination of the products compared to identical experiments with noncomplexed peptides was used to probe interactions of specific amino acids with the DNA. The ability to observe these complexes by mass spectrometry and to probe the specific interactions involved opens the door for utilizing this analytical technique to other structural biological problems including the study of transcription processes and determining the specific binding regions between dsDNA and proteins. PMID- 10821650 TI - Influence of the denticity of ligand systems on the in vitro and in vivo behavior of (99m)Tc(I)-tricarbonyl complexes: a hint for the future functionalization of biomolecules. AB - Functionalization of biologically relevant molecules for the labeling with the novel fac-[(99m)Tc(OH(2))(3)(CO)(3)](+) precursor has gained considerable attention recently. Therefore, we tested seven different tridentate (histidine L(1)(), iminodiacetic acid L(2)(), N-2-picolylamineacetic acid L(3)(), N, N-2 picolylaminediacetic acid L(4)()) and bidentate (histamine L(5)(), 2-picolinic acid L(6)(), 2,4-dipicolinic acid L(7)()) ligand systems, with the potential to be bifunctionalized and attached to a biomolecule. The ligands allowed mild radiolabeling conditions with fac-[(99m)Tc(OH(2))(3)(CO)(3)](+) (30 min, 75 degrees C). The ligand concentrations necessary to obtain yields of >95% of the corresponding organometallic complexes 1-7 ranged from 10(-)(6) to 10(-)(4) M. Complexes of the general formula "fac-[(99m)TcL(CO)(3)]" (L = tridentate ligand) and "fac-[(99m)Tc(OH(2))L'(CO)(3)]" (L' = bidentate ligand), respectively, were produced. Challenge studies with cysteine and histidine revealed significant displacement of the ligands in complexes 5-7 but only little exchange with complexes 1-4 after 24 h at 37 degrees C in PBS buffer. However, no decomposition to (99m)TcO(4)(-) was observed under these conditions. All complexes showed a hydrophilic character (log P(o/w) values ranging from -2.12 to 0.32). Time dependent FPLC analyses of compounds 1-7 incubated in human plasma at 37 degrees C showed again no decomposition to (99m)TcO(4)(-) after 24 h at 37 degrees C. However, the complexes with bidentate ligands (5-7) became almost completely protein bound after 60 min, whereas the complexes with tridentate coordinated ligands (1-4) showed no reaction with serum proteins. The compounds were tested for their in vivo stability and the biodistribution characteristics in BALB/c mice. The complexes with tridentate coordinated ligand systems (1-4) revealed generally a good and fast clearance from all organs and tissues. On the other hand, the complexes with only bidentate coordinated ligands (5-7) showed a significantly higher retention of activity in the liver, the kidneys, and the blood pool. Detailed radiometric analyses of murine plasma samples, 30 min p.i. of complex fac-[(99m)TcL(1)(CO)(3)], 1, revealed almost no reaction of the radioactive complex with the plasma proteins. By contrast, in plasma samples of mice, which were injected with complex fac-[(99m)Tc(OH(2))L(5)(CO)(3)](+), 5, the entire radioactivity coeluded with the proteins. On the basis of these in vitro and in vivo experiments, it appears that functionalization of biomolecules with tridentate-chelating ligand systems is preferable for the labeling with fac [(99m)Tc(OH(2))(3)(CO)(3)](+), since this will presumably result in radioactive bioconjugates with better pharmacokinetic profiles. PMID- 10821651 TI - Design consideration and probes for fluorescence resonance energy transfer studies. AB - Spectroscopic properties of two newly synthesized water-soluble thiol-reactive fluorescent probes, 7-(iodoacetamido)-coumarin-4-carboxylic acid (I-Cca) and N iodoacetyl-beta-(2-naphthyl)alanine (I-Nal), were characterized using single cysteine mutants of Escherichia coli adenylate kinase. Together with two known water-soluble thiol-reactive dyes (Lucifer yellow iodoacetamide and 5 iodoacetamidosalicylic acid) and as well, tryptophan residues (either native or inserted into a protein by site directed mutagenesis), these probes can be arranged pairwise in a molecular tool set for studies of structural transitions in proteins by means of fluorescence resonance energy-transfer (FRET) experiments. A set of seven donor/acceptor pairs which allow determination of intramolecular distances and their distributions over the range 10-40 A in labeled protein derivatives is described. The charged groups present in the probes facilitate the conjugation reaction and improve postlabeling purification. General considerations for design of charged probes and site-directed labeling for applications of FRET methods in studies of protein structure and dynamics are presented. PMID- 10821652 TI - Self-assembled peptides exposing epitopes recognizable by human lymphoma cells. AB - A bifunctional N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer containing nitrilotriacetic acid (NTA) and benzophenone (BP) groups was synthesized by free radical copolymerization of HPMA, 2-methacrylamidobutyl nitrilotriacetic acid (MABNTA), and 4-methacrylamido benzophenone (MABP) using 2, 2' azobisisobutyronitrile (AIBN) as initiator. A His-tagged coiled coil stem loop peptide containing a tridecapeptide (TDP) epitope (GFLGEDPGFFNVE) in the loop region (CCSL-TDP) was designed and synthesized genetically by expressing an artificial gene in Escherichia coli BL21 (DE3). The peptide was characterized by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), size-exclusion chromatography (SEC), and circular dichroism (CD) spectroscopy. Surfaces containing self-assembled CCSL-TDP peptide were prepared by first covalently grafting poly(HPMA-co-MABNTA-co-MABP) onto polystyrene (PS) surface by UV irradiation, then charging the surface with nickel through NTA groups, and finally attaching the CCSL-TDP peptide through Ni-histidine chelation. The modified PS surfaces with and without self-assembled CCSL-TDP peptide were characterized by X-ray photoelectron spectroscopy (XPS) and time-of flight secondary ion mass spectrometry (TOF-SIMS). Cell attachment studies with human Burkitt's lymphoma Raji B cells showed that the cells selectively bound to the self-assembled CCSL-TDP peptide surfaces, but not to the surfaces of PS, PS with grafted copolymer, and PS with grafted copolymer and self-assembled coiled coil peptide with similar structure but without the epitope. This indicates that the cell attachment was mediated by the CCSL-TDP peptide, most probably by the TDP epitope region. The CCSL peptide self-assembly presented here may represent a feasible model of exposing epitopes for biorecognition studies. PMID- 10821653 TI - Poly(ethylene glycol)-modification of the phospholipid vesicles by using the spontaneous incorporation of poly(ethylene glycol)-lipid into the vesicles. AB - The critical micelle concentrations of 1, 2-dipalmitoyl-sn-glycero-3 phosphoethanolamine-N-[monomethoxy poly(ethylene glycol) (5000)] (PEG-DPPE) and its distearoyl analogue (PEG-DSPE) were 70 and 9 microM, respectively, in buffer solutions ([Tris] = 20 mM, [NaCl] = 140 mM, pH 7.4) at 37 degrees C. When these PEG-lipid micelle dispersions were mixed with the dispersions of phospholipid vesicles comprised of a C16 membrane, of which the carbon number is 16, or a C18 membrane, the PEG-lipid micelles were dissociated into monomers and then spontaneously incorporated into the surface of the preformed vesicles. The incorporation rates and the enthalpy changes during incorporation were measured with an isothermal titration microcalorimeter. The incorporation rate of PEG-DPPE was faster than that of PEG-DSPE, because the dissociation rate of the PEG-DPPE micelles was faster than that of PEG-DSPE micelles. The incorporation equilibrium constant of PEG-DSPE was larger than that of PEG-DPPE due to its slow dissociation rate from the membrane, caused by the stronger hydrophobic interaction. The combination of PEG-DSPE and the C18 membrane was the most thermodynamically stabilized pair. Furthermore, the dispersion stability of the surface-modified vesicles prepared by this spontaneous incorporation was analyzed by using the critical molecular weight of the polymer for the aggregation of vesicles. The aggregation of the vesicles was successfully supressed with an increase in the molecular weight of the PEG in the PEG-lipid and its incorporation ratio. PMID- 10821654 TI - Abeta(1-40) peptide radiopharmaceuticals for brain amyloid imaging: (111)In chelation, conjugation to poly(ethylene glycol)-biotin linkers, and autoradiography with Alzheimer's disease brain sections. AB - The amyloid plaques of Alzheimer's disease (AD) are formed by the neuropeptide Abeta(1)(-)(42/43), and carboxyl terminal truncated forms of this neuropeptide, designated Abeta(1)(-)(40), bind to amyloid plaques of AD autopsy tissue sections. Therefore, Abeta(1)(-)(40) is a potential peptide radiopharmaceutical that could be used for imaging brain amyloid in living subjects with AD, should this neuropeptide be made transportable through the blood-brain barrier (BBB). To accomplish this, the neuropeptide must be modified to enable (i) attachment to a BBB drug targeting system and (ii) labeling with a radionuclide, e.g., 111 indium, suitable for brain imaging by external detection modalities such as single photon emission computed tomography (SPECT). The present studies describe the synthesis of an Abeta(1)(-)(40) analogue that contains a biotin at the amino terminus and a diethylenetriaminepentaacetic acid (DTPA) moiety conjugated to one of the internal lysine residues. The DTPA-[N-biotin]-Abeta(1)(-)(40) was purified by gel filtration fast-protein liquid chromatography (FPLC) using two Superose 12HR columns in series, and the structure of the purified peptide was confirmed by matrix-assisted laser desorption ionization (MALDI) mass spectrometry. The binding of the [(111)In]DTPA-[N-biotin]-Abeta(1)(-)(40) to amyloid plaques of AD autopsy tissue sections was demonstrated by film and emulsion autoradiography. A poly(ethylene glycol) (PEG) linker of 3400 Da molecular mass, designated PEG(3400), was inserted between the Abeta(1)(-)(40) and the biotin moiety, but this modification diminishes binding of Abeta(1)(-)(40) to the AD amyloid plaques. In summary, these studies describe a novel formulation of biotinylated Abeta(1)(-)(40) that allows radiolabeling with 111-indium. The peptide radiopharmaceutical may be conjugated to an avidin-based BBB drug targeting system to enable transport through the BBB and imaging of brain amyloid in vivo. PMID- 10821655 TI - B-Domain deleted recombinant coagulation factor VIII modified with monomethoxy polyethylene glycol. AB - Recombinant coagulation factor VIII (r-VIII SQ) was chemically modified with monomethoxy poly(ethylene glycol) (mPEG). Three mPEG derivatives were used for coupling to the r-VIII SQ lysines, a mixed anhydride of monomethoxy poly(ethylene glycol) succinic acid (mPEG-SAH), monomethoxy poly(ethylene glycol) succinimidyl succinate (mPEG-SS), and monomethoxy poly(ethylene glycol) tresylate (mPEG-TRES). A consequence of the modification with all derivatives was a substantial reduction in coagulant activity, even at very low degrees of modification. A method was developed with the purpose of avoiding conjugation at certain important biological sites on the factor VIII and thereby producing conjugates with better retained activity. This was achieved by immobilizing the protein onto a solid matrix during the modification reaction. Characterization of conjugates by SDS-PAGE, western blots, interaction with von Willebrand factor (vWf), and thrombin activation/inactivation analyses was undertaken. The SDS-PAGE and western blots revealed coupling heterogeneity regarding degree of modification. The amount of factor VIII able to bind to vWf decreased with the conjugation. Thrombin activated the modified factor VIII to essentially the same extent as the reference preparation of r-VIII SQ. Inactivation of the modified factor VIII was, however, slower than inactivation of the unmodified protein. Finally, an in vitro study was performed to evaluate the influence of the mPEG modification on the protein stability in extract of porcine tissue. Despite that conjugates with low degrees of modification were included in the study, the coagulant activity was preserved to a significantly higher extent in all incubation mixtures containing conjugates compared to that with unmodified protein. PMID- 10821656 TI - Enhancing the "stickiness" of bile acids to cross-linked polymers: a bioconjugate approach to the design of bile acid sequestrants. AB - A series of cross-linked polymers have been synthesized by reaction of 2% cross linked, chloromethylated polystyrene with a tertiary amine derivative (7) of cholic acid (prepared by reaction of cholic acid methyl ester with N,N-dimethyl ethylenediamine), followed by exhaustive quaternization with trimethylamine. Increased loadings of 7 result in enhanced binding of taurocholate ion and a decrease in its rate of release upon exposure to 50 mM aqueous NaCl. Examination of particle-size effects indicates that film diffusion and particle diffusion both contribute to the observed kinetics. Interruption tests that were carried out confirmed that particle diffusion is kinetically important. The relevance of this bioconjugate approach to the design of bile acid sequestrants for the treatment of hypercholesterolemia is briefly discussed. PMID- 10821657 TI - Targeting hypoxia in tumors using 2-nitroimidazoles with peptidic chelators for technetium-99m: effect of lipophilicity. AB - Tumor hypoxia is an important prognostic factor for response to therapy. Radiolabeled 2-nitroimidazoles have been used for imaging hypoxia, and the octanol/water partition coefficient (P) of these compounds appears to play a crucial role in their suitability for imaging. A series of 11 2-nitroimidazoles coupled to peptidic chelators for (99m)Tc with divergent P was developed and evaluated in an in vitro system. Two classes of N(3)S chelators were used: dialkyl-Gly-Ser-Cys-linker-2-nitroimidazole (Class I) and dialkyl-Gly-Lys(2 nitroimidazole)-Cys (Class II). The chelators were prepared by automated solid phase peptide synthesis. Xanthine oxidase was able to reduce the 2 nitroimidiazole moiety on the ligands, but the rate of reduction varied 5-fold among the different chelators. The chelators were labeled by transchelation from [(99m)Tc]gluconate at temperatures between 22 and 100 degrees C. The reaction mixtures were analyzed by HPLC and their P values determined. The accumulation of each complex in suspension cultures of Chinese hamster ovary cells incubated under aerobic or extremely hypoxic conditions was determined. Radiochemical yields ranged from 5 to 80% for the 11 compounds. HPLC showed that some of the compounds formed two complexes with (99m)Tc, possibly syn and anti conformations with respect to the Tc=O bond. In general, the Class I chelators labeled more readily than the class II chelators. The P values of the (99m)Tc complexes varied from 0.0002 to 5 and were generally in accordance with predictions based on structure. There were also differences in P as a function of pH; the free acids had a lower P at pH 7.4 than at pH 2.0 due to ionization, whereas the amides did not show this effect. Accumulation levels in aerobic cells were related to P but varied over a narrow range. Four of the 11 compounds showed selective accumulation in hypoxic cells. The peptidic class of 2-nitroimidazoles, with flexible design and convenient solid-phase synthesis, deserves further study as agents for imaging hypoxia in tumors. PMID- 10821658 TI - Sulfur shuffle: modulating enzymatic activity by thiol-disulfide interchange. AB - The facile modulation of biological processes is an important goal of biological chemists. Here, a general strategy is presented for controlling the catalytic activity of an enzyme. This strategy is demonstrated with ribonuclease A (RNase A), which catalyzes the cleavage of RNA. The side-chain amino group of Lys41 donates a hydrogen bond to a nonbridging oxygen in the transition state for RNA cleavage. Replacing Lys41 with a cysteine residue is known to decrease the value of k(cat)/K(m) by 10(5)-fold. Forming a mixed disulfide between the side chain of Cys41 of K41C RNase A and cysteamine replaces the amino group and increases k(cat)/K(m) by 10(3)-fold. This enzyme, which contains a mixed disulfide, is readily deactivated by dithiothreitol. Forming a mixed disulfide between the side chain of Cys41 and mercaptopropyl phosphate, which is designed to place a phosphoryl group in the active site, decreases activity by an additional 25-fold. This enzyme, which also contains a mixed disulfide, is reactivated in the presence of dithiothreitol and inorganic phosphate (which displaces the pendant phosphoryl group from the active site). An analogous control mechanism could be installed into the active site of virtually any enzyme by replacing an essential residue with a cysteine and elaborating the side chain of that cysteine into appropriate mixed disulfides. PMID- 10821659 TI - Chemical and biological characterization of technetium(I) and Rhenium(I) tricarbonyl complexes with dithioether ligands serving as linkers for coupling the Tc(CO)(3) and Re(CO)(3) moieties to biologically active molecules. AB - The organometallic precursor (NEt(4))(2)[ReBr(3)(CO)(3)] was reacted with bidendate dithioethers (L) of the general formula H(3)C-S-CH(2)CH(2)-S-R (R = CH(2)CH(2)COOH, CH(2)-C&tbd1;CH) and R'-S-CH(2)CH(2)-S-R' (R' = CH(3)CH(2)-, CH(3)CH(2)-OH, and CH(2)COOH) in methanol to form stable rhenium(I) tricarbonyl complexes of the general composition [ReBr(CO)(3)L]. Under these conditions, the functional groups do not participate in the coordination. As a prototypic representative of this type of Re compounds, the propargylic group bearing complex [ReBr(CO(3))(H(3)C-S-CH(2)CH(2)-S-CH(2)C&tbd1;CH)] Re2 was studied by X ray diffraction analysis. Its molecular structure exhibits a slightly distorted octahedron with facial coordination of the carbonyl ligands. The potentially tetradentate ligand HO-CH(2)CH(2)-S-CH(2)CH(2)-S-CH(2)CH(2)-OH was reacted with the trinitrato precursor [Re(NO(3))(3)(CO)(3)](2-) to yield a cationic complex [Re(CO)(3)(HO-CH(2)CH(2)-S-CH(2)CH(2)-S-CH(2)CH(2)-OH)]NO(3) Re8 which shows the coordination of one hydroxy group. Re8 has been characterized by correct elemental analysis, infrared spectroscopy, capillary electrophoresis, and X-ray diffraction analysis. Ligand exchange reaction of the carboxylic group bearing ligands H(3)C-S-CH(2)CH(2)-S-CH(2)CH(2)-COOH and HOOC-CH(2)-S-CH(2)CH(2)-S-CH(2) COOH with (NEt(4))(2)[ReBr(3)(CO)(3)] in water and with equimolar amounts of NaOH led to complexes in which the bromide is replaced by the carboxylic group. The X ray structure analysis of the complex [Re(CO)(3)(OOC-CH(2)-S-CH(2)CH(2)-S-CH(2) COOH)] Re6 shows the second carboxylic group noncoordinated offering an ideal site for functionalization or coupling a biomolecule. The no-carrier-added preparation of the analogous (99m)Tc(I) carbonyl thioether complexes could be performed using the precursor fac-[(99m)Tc(H(2)O)(3)(CO)(3)](+), with yields up to 90%. The behavior of the chlorine containing (99m)Tc complex [(99m)TcCl(CO)(3)(CH(3)CH(2)-S-CH(2)CH(2)-S-CH(2)CH(3))] Tc1 in aqueous solution at physiological pH value was investigated. In saline, the chromatographically separated compound was stable for at least 120 min. However, in chloride-free aqueous solution, a water-coordinated cationic species Tc1a of the proposed composition [(99m)Tc(H(2)O)(CO)(3)(CH(3)CH(2)-S-CH(2)CH(2)-S-CH(2)CH(3))](+) occurred. The cationic charge of the conversion product was confirmed by capillary electrophoresis. By the introduction of a carboxylic group into the thioether ligand as a third donor group, the conversion could be suppressed and thus the neutrality of the complex preserved. Biodistribution studies in the rat demonstrated for the neutral complexes [(99m)TcCl(CO)(3)(CH(3)CH(2)-S-CH(2)CH(2) S-CH(2)CH(3))] Tc1 and [(99m)TcCl(CO)(3)(CH(2)-S-CH(2)CH(2)-S-CH(2)-C&tbd1;CH)] Tc2 a significant initial brain uptake (1.03 +/- 0.25% and 0.78 +/- 0.08% ID/organ at 5 min. p.i.). Challenge experiments with glutathione clearly indicated that no transchelation reaction occurs in vivo. PMID- 10821660 TI - Poly(ethylene glycol)-conjugation and deoxygenation enable long-term preservation of hemoglobin-vesicles as oxygen carriers in a liquid state. AB - The stability of hemoglobin vesicles (HbV) as an oxygen infusion was tested during the storage for 1 year at 4, 23, and 40 degrees C. The surface of the HbV was modified with poly(ethylene glycol) (PEG), and the suspension was deoxygenated with nitrogen bubbling. The samples stored at 4 and 23 degrees C showed a stable dispersion state for 1 year, though the sample stored at 40 degrees C showed the precipitation and decomposition of vesicular components, a decrease in pH, and 4% leakage of total Hb after 1 year. The PEG chains on the vesicular surface stabilize the dispersion state and prevent the aggregation and fusion due to their steric hindrance. The original metHb content (ca. 3%) before the preservation gradually decreased to less than 1% in all the samples after 1 month due to the presence of homocysteine inside the vesicles which consumed the residual oxygen and gradually reduced the trace amount of metHb. The rate of metHb formation was strongly dependent on the partial pressure of oxygen, and no increase in metHb formation was observed due to the intrinsic stability of the deoxygenated Hb. Preservation at 4 and 23 degrees C slightly reduced P(50) (increased the oxygen affinity) from 38 Torr to 32 and 31 Torr, respectively. These results indicate the possibility that HbV suspension can be stored at room temperature for at least 1 year. PMID- 10821662 TI - Synthesis, spectral properties, and detection limits of reactive squaraine dyes, a new class of diode laser compatible fluorescent protein labels PMID- 10821661 TI - Fluorescent-labeled poly(ethylene glycol) lipid conjugates with distal cationic headgroups. AB - The synthesis of a new class of fluorescent cationic poly(ethylene glycol) lipid conjugates (CPLs) is described. These lipids consist of a hydrophobic distearoyl phosphatidylethanolamine (DSPE) anchor coupled to a highly fluorescent N(epsilon) dansyl lysine moiety, which is attached to a hydrophilic poly(ethylene glycol) (PEG) spacer that is linked to a cationic headgroup made of lysine residues. Introduction of the dansyl moiety allows rapid and accurate quantification of CPLs within lipid bilayers using fluorescence techniques. The synthetic scheme is straightforward, using repeated amino-carboxyl coupling reaction steps, with purification by precipitation. A series of dansylated CPLs was synthesized with zero, one, three, and seven lysine residues located at the distal end of the PEG chain, giving rise to CPLs with one, two, four, and eight distal positive charges, respectively. The structures of the CPLs were confirmed by (1)H NMR spectroscopy and chemical analysis. CPLs provide a means of introducing positive charge to a bilayer that is localized some distance from the membrane surface, and are of particular interest for nonviral gene delivery applications. The usefulness of CPLs is demonstrated by the enhanced in vitro cellular binding and uptake of liposomes containing CPL(4). PMID- 10821663 TI - Reaction of bovine cytochrome c oxidase with hydrogen peroxide produces a tryptophan cation radical and a porphyrin cation radical. AB - Oxidized bovine cytochrome c oxidase reacts with hydrogen peroxide to generate two electron paramagnetic resonance (EPR) free radical signals (Fabian, M., and Palmer, G. (1995) Biochemistry 34, 13802-13810). These radicals are associated with the binuclear center and give rise to two overlapped EPR signals, one signal being narrower in line width (DeltaHptp = 12 G) than the other (DeltaHptp = 45 G). We have used electron nuclear double resonance (ENDOR) spectrometry to identify the two different chemical species giving rise to these two EPR signals. Comparison of the ENDOR spectrum associated with the narrow signal with that of compound I of horseradish peroxidase (formed by reaction of that enzyme with hydrogen peroxide) demonstrates that the two species are virtually identical. The chemical species giving rise to the narrow signal is therefore identified as an exchange-coupled porphyrin cation radical similar to that formed in horseradish peroxidase compound I. Comparison of the ENDOR spectrum of compound ES (formed by the reaction of hydrogen peroxide with cytochrome c peroxidase) with that of the broad signal indicates that the chemical species giving rise to the broad EPR signal in cytochrome c oxidase is probably an exchange coupled tryptophan cation radical. This is substantiated using H(2)O/D(2)O solvent exchange experiments where the ENDOR difference spectrum of the broad EPR signal of cytochrome c oxidase shows a feature consistent with hyperfine coupling to the exchangeable N(1) proton of a tryptophan cation radical. PMID- 10821664 TI - Elucidation of distinct ligand binding sites for cytochrome P450 3A4. AB - Cytochrome P450 (P450) 3A4 is the most abundant human P450 enzyme and has broad selectivity for substrates. The enzyme can show marked catalytic regioselectivity and unusual patterns of homotropic and heterotropic cooperativity, for which several models have been proposed. Spectral titration studies indicated one binding site for the drug indinavir (M(r) 614), a known substrate and inhibitor. Several C-terminal aminated peptides, including the model morphiceptin (YPFP NH(2)), bind with spectral changes indicative of Fe-NH(2) bonding. The binding of the YPFP-NH(2) N-terminal amine and the influence of C-terminal modification on binding argue that the entire molecule (M(r) 521) fits within P450 3A4. YPFP NH(2) was not oxidized by P450 3A4 but blocked binding of the substrates testosterone and midazolam, with K(i) values similar to the spectral binding constant (K(s)) for YPFP-NH(2). YPFP-NH(2) inhibited the oxidations of several typical P450 substrates with K(i) values 10-fold greater than the K(s) for binding YPFP-NH(2) and its K(i) for inhibiting substrate binding. The n values for cooperativity of these oxidations were not altered by YPFP-NH(2). YPFP-NH(2) inhibited the oxidations of midazolam at two different positions (1'- and 4-) with 20-fold different K(i) values. The differences in the K(i) values for blocking the binding to ferric P450 3A4 and the oxidation of several substrates may be attributed to weaker binding of YPFP-NH(2) to ferrous P450 3A4 than to the ferric form. The ferrous protein can be considered a distinct form of the enzyme in binding and catalysis because many substrates (but not YPFP-NH(2)) facilitate reduction of the ferric to ferrous enzyme. Our results with these peptides are considered in the context of several proposed models. A P450 3A4 model based on these peptide studies contains at least two and probably three distinct ligand sites, with testosterone and alpha-naphthoflavone occupying distinct sites. Midazolam appears to be able to bind to P450 3A4 in two modes, one corresponding to the testosterone binding mode and one postulated to reflect binding in a third site, distinct from both testosterone and alpha-naphthoflavone. The work with indinavir and YPFP-NH(2) also argues that room should be present in P450 3A4 to bind more than one smaller ligand in the "testosterone" site, although no direct evidence for such binding exists. Although this work with peptides provides evidence for the existence of multiple ligand binding sites, the results cannot be used to indicate their juxtaposition, which may vary through the catalytic cycle. PMID- 10821665 TI - Modeling the effects of mutations on the free energy of the first electron transfer from QA- to QB in photosynthetic reaction centers. AB - Numerical calculations of the free energy of the first electron transfer in genetically modified reaction centers from Rhodobacter (Rb.) sphaeroides and Rb. capsulatus were carried out from pH 5 to 11. The multiconformation continuum electrostatics (MCCE) method allows side chain, ligand, and water reorientation to be embedded in the calculations of the Boltzmann distribution of cofactor and amino acid ionization states. The mutation sites whose effects have been modeled are L212 and L213 (the L polypeptide) and two in the M polypeptide, M43(44) and M231(233) in Rb. capsulatus (Rb. sphaeroides). The results of the calculations were compared to the experimental data, and very good agreement was found especially at neutral pH. Each mutation removes or introduces ionizable residues, but the protein maintains a net charge close to that in native RCs through ionization changes in nearby residues. This reduces the effect of mutation and makes the changes in state free energy smaller than would be found in a rigid protein. The state energy of QA-QB and QAQB- states have contributions from interactions among the residues as well as with the quinone which is ionized. For example, removing L213Asp, located in the QB pocket, predominantly changes the free energy of the QA-QB state, where the Asp is ionized in native RCs rather than the QAQB- state, where it is neutral. Side chain, hydroxyl, and water rearrangements due to each of the mutations have also been calculated showing water occupancy changes during the QA- to QB electron transfer. PMID- 10821666 TI - Structural consequences of the replacement of glycine M203 with aspartic acid in the reaction center from Rhodobacter sphaeroides. AB - Reaction centers with the double mutation Phe M197 to Arg and Gly M203 to Asp (FM197R/GM203D) have been crystallized from an antenna-deficient strain of Rhodobacter sphaeroides, and the structure has been determined at 2.7 A resolution. Unlike in reaction centers with a single FM197R mutation, the Arg M197 residue in the FM197R/GM203D reaction center adopts a position similar to that of the native Phe residue in the wild-type reaction center. Asp M203 is packed in such a way that the gamma-carboxy group interacts with the backbone carbonyl of Arg M197. The Asp M203 residue takes up part of the volume that is occupied in the wild-type reaction center by a water molecule. This water has been proposed to form a hydrogen bond interaction with the 9-keto carbonyl group of the active branch accessory bacteriochlorophyll, particularly when the primary donor bacteriochlorophylls are oxidized. The GM203D mutation therefore appears to remove the possibility of this hydrogen bond interaction by exclusion of this water molecule, as well as altering the local dielectric environment of the 9 keto carbonyl group. We examine whether the observed structural changes can provide new or alternative explanations for the absorbance and electron-transfer properties of reaction centers with the FM197R and GM203D mutations. PMID- 10821667 TI - Functions of fluctuation in the heme-binding loops of cytochrome b5 revealed in the process of heme incorporation. AB - Cytochrome b(5) (cyt b(5)) holds heme using two axial histidines, His63 and His39, that are located in the centers of the two heme-binding loops. The previous NMR study on the apo form of cyt b(5) (apocyt b(5)) revealed that the loop including His63 exhibits a larger fluctuation compared to the other loop including His39 [Falzone, C. J., Mayer, M. R., Whiteman, E. L., Moore, C. D., and Lecomte, J. T. (1996) Biochemistry 35, 6519-6526]. To understand the significance of the fluctuation, the heme association and dissociation rates of the two loops were compared using two mutants of cyt b(5) in which one of the axial histidines was replaced with leucine. It was demonstrated that the fluctuating loop possesses a significantly slower heme dissociation rate and a faster heme association rate than the other loop. To further verify the importance of the fluctuating loop, the heme association process of wild-type apocyt b(5) was investigated using optical absorption and CD spectroscopies. It was indicated that the process proceeds through the two pathways, and that the dominant pathway involves the initial coordination of His63 located in the fluctuating loop. The urea concentration dependency of the rate constants revealed that the folding of the fluctuating loop is associated with the coordination of His63. It was suggested that the fluctuation enables the loop to have a larger heme-loop contact in the heme-bound conformation. The fluctuating heme-binding loops might be useful for the artificial design of heme-binding proteins. PMID- 10821668 TI - Tryptophan-heme pi-electrostatic interactions in cytochrome f of oxygenic photosynthesis. AB - Cytochrome f of oxygenic photosynthesis has an unprecedented structure, including the N-terminus being a heme ligand. The adjacent N-terminal heme-shielding domain is enriched in aromatic amino acids. The atomic structures of the chloroplast and cyanobacterial cytochromes f were compared to explain spectral and redox differences between them. The conserved aromatic side chain in the N-terminal heme-shielding peptide at position 4, Phe and Tyr in plants and algae, respectively, and Trp in cyanobacteria, is in contact with the heme. Mutagenesis of cytochrome f from the eukaryotic green alga Chlamydomonas reinhardtii showed that a Phe4 --> Trp substitution in the N-terminal domain was unique in causing a red shift of 1 and 2 nm in the cytochrome Soret (gamma) and Q (alpha) visible absorption bands, respectively. The resulting alpha band peak at 556 nm is characteristic of the cyanobacterial cytochrome. Conversely, a Trp4 --> Phe mutation in the expressed cytochrome from the cyanobacterium Phormidium laminosum caused a blue shift to the 554 nm alpha band peak diagnostic of the chloroplast cytochrome. Residue 4 was found to be the sole determinant of this 60 cm(-)(1) spectral shift, and of approximately one-half of the 70 mV redox potential difference between cytochrome f of P. laminosum and C. reinhardtii (E(m7) = 297 and 370 mV, respectively). The proximity of Trp-4 to the heme implies that the spectral and redox potential shifts arise through differential interaction of its sigma- or pi-electrostatic potential with the heme ring and of the pi-potential with the heme Fe orbitals, respectively. The dependence of the visible spectrum and redox potential of cytochrome f on the identity of aromatic residue 4 provides an example of the use of the relatively sharp cytochrome spectrum as a "spectral fingerprint", and of the novel structural connection between the heme and a single nonliganding residue. PMID- 10821669 TI - Monomeric midkine induces tumor cell proliferation in the absence of cell-surface proteoglycan binding. AB - Midkine (MK), a retinoic acid-inducible heparin-binding protein, is a mitogen which initiates a cascade of intracellular protein tyrosine phosphorylation mediated by the JAK/STAT pathway after binding to its high affinity p200(+)/MKR cell surface receptor in the G401 cell line [Ratovitski, E. A. (1998) J. Biol. Chem. 273, 3654-3660]. In this study, we determined the biophysical characteristics of purified recombinant murine MK and analyzed the requirements for ligand multimerization and cell surface proteoglycan binding for the G401 cell mitogenic activity of MK. Our studies indicate that the secreted form of MK (M = 13 kDa) exists in solution as an asymmetric monomer with a frictional coefficient of 1. 48 and a Stokes radius of 23.7 A. By constructing bead models of MK using the program AtoB and the program HYDRO to predict the hydrodynamic properties of each model, our data suggest that MK has a dumb-bell shape in solution composed of independent N- and C-terminal domains separated by an extended linker. This asymmetric MK monomer is a biologically active ligand with mitogenic activity on G401 cells in vitro. Neither heparin-induced formation of noncovalent MK multimers nor tissue transglutaminase II covalent multimerization of MK enhanced MK mitogenic activity in this system. Since neither heparin competition nor cell treatment with chondroitinase ABC or heparinase III abolished the mitogenic effects of MK on G401 cells, cell-surface proteoglycan binding by MK does not appear to be a requirement for its observed mitogenic effects. These results provide strong evidence that the MK-specific p200(+)/MKR has distinctive biochemical properties which distinguish it from the receptor tyrosine phosphatase cell-surface proteoglycan PTPzeta/RPTPbeta and support the hypothesis that the diverse biological effects of MK are mediated by multiple cell-specific signal transduction receptors. PMID- 10821670 TI - Fibril formation and neurotoxicity by a herpes simplex virus glycoprotein B fragment with homology to the Alzheimer's A beta peptide. AB - Despite significant progress in the elucidation of the genetic basis of early onset familial Alzheimer's disease (AD), the etiology of sporadic cases remains elusive. Although certain genetic loci play a role in conferring susceptibility in some sporadic AD cases, it is likely that the etiology is multifactorial; hence, the majority of cases cannot be attributed to genetic factors alone, indicating that environmental factors may modulate the onset and/or progression of the disease. Head injury and infectious agents are environmental factors that have been periodically implicated, but no plausible mechanisms have been clearly identified. With regard to infectious agents, speculation has often centered on the neurotropic herpes viruses, with herpes simplex virus 1 (HSV1) considered a likely candidate. We report that an internal sequence of HSV1 glycoprotein B (gB) is homologous to the carboxyl-terminal region of the A beta peptide that accumulates in diffuse and neuritic plaques in AD. Synthetic peptides were generated and the biophysical and biological properties of the viral peptide compared to those of A beta. Here we show that this gB fragment forms beta pleated sheets, self-assembles into fibrils that are thioflavin-positive and ultrastructurally indistinguishable from A beta, accelerates the formation of A beta fibrils in vitro, and is toxic to primary cortical neurons at doses comparable to those of A beta. These findings suggest a possible role for this infectious agent in the pathophysiology of sporadic cases of AD. PMID- 10821671 TI - Inhibition of human telomerase by rubromycins: implication of spiroketal system of the compounds as an active moiety. AB - We found that a group of rubromycins and their analogues, a class of quinone antibiotics that possesses benzofuran and benzodipyran rings to form a spiroketal system, strongly inhibited human telomerase as assessed with a modified telomeric repeat amplification protocol. beta- and gamma-Rubromycins and purpuromycin appeared to be the most potent telomerase inhibitors, with 50% inhibitory concentrations (IC(50)) of about 3 microM, and griseorhodins A and C also showed comparable potencies for the inhibition (IC(50) = 6-12 microM). In contrast, opening of the spiroketal system of beta-rubromycin, giving rise to alpha rubromycin, substantially decreased its inhibitory potency toward telomerase (IC(50) > 200 microM), indicating the essential role of the spiroketal system in telomerase inhibition. A kinetic study of the inhibition by beta-rubromycin revealed a competitive interaction with respect to the telomerase substrate primer, with a K(i) of 0.74 microM, whereas a mixed type inhibition was observed with respect to the nucleotide substrate. beta-Rubromycin was also potent in inhibiting retroviral reverse transcriptases but had virtually no effect on other DNA/RNA-modifying enzymes including DNA and RNA polymerases, deoxyribonuclease, and topoisomerase. Although beta-rubromycin showed nonspecific cytotoxicities, reducing proliferation of cancer cells (IC(50) approximately 20 microM), we conclude that beta-rubromycin appears to be a lead structure for the development of more potent and selective inhibitors of human telomerase. PMID- 10821672 TI - Rational design of femtomolar inhibitors of isoleucyl tRNA synthetase from a binding model for pseudomonic acid-A. AB - This paper describes the design and characterization of novel inhibitors of IleRS, whose binding affinity approaches the tightest reported for noncovalent inhibition. Compounds were designed from a binding model for the natural product pseudomonic acid-A (PS-A) together with a detailed understanding of the reaction cycle of IleRS and characterization of the mode of binding of the reaction intermediate IleAMP. The interactions of the compounds with IleRS were characterized by inhibition of aminoacylation of tRNA or PP(i)/ATP exchange at supersaturating substrate concentration and by transient kinetics and calorimetry methods. A detailed understanding of the interaction of a comprehensive series of compounds with IleRS allowed the identification of key features and hence the design of exquisitely potent inhibitors. Predictions based on these results have been recently supported by a docking model based on the crystal structure of IleRS with PS-A [Silvian, L. F., Wang J. M., and Steitz T. A. (1999) Science 285 1074-1077]. PMID- 10821673 TI - Effects of the N-linked glycans on the 3D structure of the free alpha-subunit of human chorionic gonadotropin. AB - To gain insight into intramolecular carbohydrate-protein interactions at the molecular level, the solution structure of differently deglycosylated variants of the alpha-subunit of human chorionic gonadotropin have been studied by NMR spectroscopy. Significant differences in chemical shifts and NOE intensities were observed for amino acid residues close to the carbohydrate chain at Asn78 upon deglycosylation beyond Asn78-bound GlcNAc. As no straightforward strategy is available for the calculation of the NMR structure of intact glycoproteins, a suitable computational protocol had to be developed. To this end, the X-PLOR carbohydrate force field designed for structure refinement was extended and modified. Furthermore, a computational strategy was devised to facilitate successful protein folding in the presence of extended glycans during the simulation. The values for phi and psi dihedral angles of the glycosidic linkages of the oligosaccharide core fragments GlcNAc2(beta1-4)GlcNAc1 and Man3(beta1 4)GlcNAc2 are restricted to a limited range of the broad conformational energy minima accessible for free glycans. This demonstrates that the protein core affects the dynamic behavior of the glycan at Asn78 by steric hindrance. Reciprocally, the NMR structures indicate that the glycan at Asn78 affects the stability of the protein core. The backbone angular order parameters and displacement data of the generated conformers display especially for the beta turn 20-23 a decreased structural order upon splitting off the glycan beyond the Asn78-bound GlcNAc. In particular, the Asn-bound GlcNAc shields the protein surface from the hydrophilic environment through interaction with predominantly hydrophobic amino acid residues located in both twisted beta-hairpins consisting of residues 10-28 and 59-84. PMID- 10821674 TI - Chemical shift mapped DNA-binding sites and 15N relaxation analysis of the C terminal KH domain of heterogeneous nuclear ribonucleoprotein K. AB - The K homology (KH) motif is one of the major classes of nucleic acid binding proteins. Some members of this family have been shown to interact with DNA while others have RNA targets. There have been no reports containing direct experimental evidence regarding the nature of KH module-DNA interaction. In this study, the interaction of the C-terminal KH domain of heterogeneous nuclear ribonucleoprotein K (KH3) with its cognate single-stranded DNA (ssDNA) are investigated. Chemical shift perturbation mapping indicates that the first two helices, the conserved GxxG loop, beta 1, and beta 2, are the primary regions involved in DNA binding for KH3. The nature of the KH3-ssDNA interaction is further illuminated by a comparison of backbone 15N relaxation data for the bound and unbound KH3. Relaxation data are also used to confirm that the backbone of wild-type KH3 is structurally identical to that of the G26R mutant KH3, which was previously published. Amide proton exchange experiments indicate that the two helices involved in DNA binding are less stable than other regions of secondary structure and that a large portion of KH3 backbone amide hydrogens are protected in some manner upon ssDNA binding. The major backbone dynamics features of KH3 are similar to those of the structurally comparable human papillomavirus-31 E2 DNA binding domain. Secondary structure information for ssDNA-bound wild-type KH3 is also presented and shows that binding results in no global changes in the protein fold. PMID- 10821675 TI - Catalytic action of fuculose 1-phosphate aldolase (class II) as derived from structure-directed mutagenesis. AB - Previous analyses established the structures of unligated L-fuculose 1-phosphate aldolase and of the enzyme ligated with an inhibitor mimicking the substrate dihydroxyacetone phosphate. These data allowed us to suggest a catalytic mechanism. On the basis of this proposal, numerous mutations were now introduced at the active center and tested with respect to their catalytic rates and their product distributions. For several mutants, the structures were determined. The results demonstrate the catalytic importance of some particular residues in defined conformations and in the mobile C-terminal chain end. Moreover, they led to a modification of the proposed mechanism. The effect of some mutations on enantioselectivity and on the ratio of diastereomer formation indicates clearly the binding site of the aldehyde moiety in relation to the other substrate dihydroxyacetone phosphate. PMID- 10821676 TI - Rod outer segment membrane guanylate cyclase type 1-linked stimulatory and inhibitory calcium signaling systems in the pineal gland: biochemical, molecular, and immunohistochemical evidence. AB - Recent evidence indicates the presence of a novel alpha(2D/A)-adrenergic receptor (alpha(2D/A)-AR) linked membrane guanylate cyclase signal transduction system in the pineal gland. This system operates via a Ca(2+)-driven rod outer segment membrane guanylate cyclase (ROS-GC). In the present study, this transduction system has been characterized via molecular, immunohistochemical, and biochemical approaches. The two main components of the system are ROS-GC1 and its Ca(2+) regulator, S100B. Both components coexist in pinealocytes where the signaling component alpha(2D/A)-AR also resides. The presence of ROS-GC2 was not detected in the pineal gland. Thus, transduction components involved in processing alpha(2D/A)-AR-mediated signals are Ca(2+), S100B, and ROS-GC1. During this investigation, an intriguing observation was made. In certain pinealocytes, ROS GC1 coexisted with its other Ca(2+) modulator, guanylate cyclase activating protein type 1 (GCAP1). In these pinealocytes, S100B was not present. The other GCAP protein, GCAP2, which is also a known modulator of ROS-GC in photoreceptors, was not present in the pineal gland. The results establish the identity of an alpha(2D/A)-AR-linked ROS-GC1 transduction system in pinealocytes. Furthermore, the findings show that ROS-GC1, in a separate subpopulation of pinealocytes, is associated with an opposite Ca(2+) signaling pathway, which is similar to phototransduction in retina. Thus, like photoreceptors, pinealocytes sense both positive and negative Ca(2+) signals, where ROS-GC1 plays a pivotal role; however, unlike photoreceptors, the pinealocyte is devoid of the ROS-GC2/GCAP2 signal transduction system. PMID- 10821677 TI - Human CC chemokine I-309, structural consequences of the additional disulfide bond. AB - I-309 is a member of the CC subclass of chemokines and is one of only three human chemokines known to contain an additional, third disulfide bond. The three dimensional solution structure of I-309 was determined by (1)H nuclear magnetic resonance spectroscopy and dynamic simulated annealing. The structure of I-309, which remains monomeric at high concentrations, was determined on the basis of 978 experimental restraints. The N-terminal region of I-309 was disordered, as has been previously observed for the CC chemokine eotaxin but not others such as MCP-1 and RANTES. This was followed in I-309 by a well-ordered region between residues 13 and 69 that consisted of a 3(10)-helix, a triple-stranded antiparallel beta-sheet, and finally a C-terminal alpha-helix. Root-mean-square deviations of 0.61 and 1.16 were observed for the backbone and heavy atoms, respectively. A comparison of I-309 to eotaxin and HCC-2 revealed a significant structural change in the C-terminal region of the protein. The alpha-helix normally present in chemokines was terminated early and was followed by a short section of extended strand. These changes were a direct result of the additional disulfide bond present in this protein. An examination of the I-309 structure will aid in an understanding of the specificity of this protein with its receptor, CCR8. PMID- 10821678 TI - Bicarbonate accelerates assembly of the inorganic core of the water-oxidizing complex in manganese-depleted photosystem II: a proposed biogeochemical role for atmospheric carbon dioxide in oxygenic photosynthesis. AB - The proposed role for bicarbonate (HCO(3)(-)) as an intrinsic cofactor within the water-oxidizing complex (WOC) of photosystem II (PSII) [Klimov et al. (1997) Biochemistry 36, 16277-16281] was tested by investigation of its influence on the kinetics and yield of photoactivation, the light-induced assembly of the functional inorganic core (Mn(4)O(y)Ca(1)Cl(x)) starting from the cofactor depleted apo-WOC-PSII center and free Mn(2+), Ca(2+), and Cl(-). Two binding sites for bicarbonate were found that stimulate photoactivation by accelerating the formation and suppressing the decay, respectively, of the first light-induced assembly intermediate, IM(1) [apo-WOC-Mn(OH)(2)(+)]. A high-affinity bicarbonate site (K(D) 25% of the known accessible potential in native proteins with axial His-Met ligation) through mutation at these two positions. The redox potential of the wild-type protein occurs at an extremum of the distribution observed, indicating that Phe61 and Phe65 were most likely naturally selected to differentially stabilize the reduced state of the protein. At the other extremum, a compositionally conservative set of mutations (F61I, F65Y) leads to a 100 mV shift in the redox equilibrium toward the oxidized state. NMR analyses indicate that a charge-dipole interaction which results from mutation of phenylalanine to tyrosine at position 65 may be responsible. PMID- 10821681 TI - Adsorption of pulmonary surfactant protein SP-A to monolayers of phospholipids containing hydrophobic surfactant protein SP-B or SP-C: potential differential role for tertiary interaction of lipids, hydrophobic proteins, and SP-A. AB - Surface balance techniques were used to study the interactions of surfactant protein SP-A with monolayers of surfactant components preformed at the air-water interface. SP-A adsorption into the monolayers was followed by monitoring the increase in the surface pressure Deltapi after injection of SP-A beneath the films. Monolayers of dipalmitoylphosphatidylcholine (DPPC):egg phosphatidylglycerol (PG) (8:2, mol/mol) spread at initial surface pressure pi(i) = 5 mN/m did not promote the adsorption of SP-A at a subphase concentration of 0.68 microg/mL as compared to its adsorption to the monolayer-free surface. Surfactant proteins, SP-B or SP-C, when present in the films of DPPC:PG spread at pi(i) = 5 mN/m, enhanced the incorporation of SP-A in the monolayers to a similar extent; the Deltapi values being dependent on the levels of SP-B or SP-C, 3-17 wt %, in the lipid films. Calcium in the subphase did not affect the intrinsic surface activity of SP-A but reduced the Deltapi values produced by the adsorption of the protein to all the preformed films independently of their compositions and charges. The divalent ions likely modified the interaction of SP A with the monolayers through their effects on the conformation, self association, and charge state of SP-A. Values of Deltapi produced by adsorption of SP-A to the films of DPPC:PG with or without SP-B or SP-C were a function of the initial surface pressure of the films, pi(i). In the range of pressures 5 Cys/Val149-->Cys was studied. Ala replacement for Arg144 causes a marked decrease in cross-linking, while Ala replacement for Glu126 alone or for both Glu126 and Arg144 has little effect. The data provide strong support for the argument that Glu126 and Arg144 are within close proximity and suggest that an unpaired carboxylate at position 126 causes a structural change at the interface between helices IV and V. PMID- 10821687 TI - In vitro polymerization of tau protein monitored by laser light scattering: method and application to the study of FTDP-17 mutants. AB - Tau polymerization into the filaments that compose neurofibrillary tangles is seminal to the development of many neurodegenerative diseases. It is therefore important to understand the mechanisms involved in this process. However, a consensus method for monitoring tau polymerization in vitro has been lacking. Here we demonstrate that illuminating tau polymerization reactions with laser light and measuring the increased scattering at 90 degrees to the incident beam with a digital camera results in data that closely approximate the mass of tau polymer formation in vitro. The validity of the technique was demonstrated over a range of tau concentrations and through multiple angle scattering measurements. In addition, laser light scattering data closely correlated with quantitative electron microscopy measurements of the mass of tau filaments. Laser light scattering was then used to measure the efficiency with which the mutant tau proteins found in frontotemporal dementia and Parkinsonism linked to chromosome 17 (FTDP-17) form filamentous structures. Several of these mutant proteins display enhanced polymerization in the presence of arachidonic acid, suggesting a direct role for these mutations in tau the filament formation that characterizes FTDP-17. PMID- 10821688 TI - Two distinct forms of MAPKAP kinase-2 in adult cardiac ventricular myocytes. AB - Hsp27 kinase activities were studied in adult rat ventricular myocytes following sequential chromatography on Mono Q and Mono S. A basal level of activity was present following cell isolation. FPLC on Mono Q revealed three peaks of activity, peaks 'a', 'b', and 'c'. A fourth peak, 'd', was detected upon subsequent chromatography of the Mono Q flow-through on Mono S. Immunoblotting revealed that peaks 'a', 'b', and 'c' contained predominantly a 49 kDa form of MAPKAP kinase-2. Peak 'd' contained a 43 kDa form. 'In-gel' kinase assays using hsp27 indicated both forms of MAPKAP kinase-2 were active. No other bands of hsp27 kinase activity were detected. Both forms of hsp27 kinase immunoprecipitated with a MAPKAP kinase-2 antibody and have therefore been named MAPKAP kinase-2alpha (p49) and MAPKAP kinase-2beta (p43). MAPKAP kinase-2beta chromatographed on Superose 12 as a 60.7 kDa monomer whereas the behavior of MAPKAP kinase-2alpha suggested both a 65.7 kDa monomer and higher molecular mass complexes. Both activities phosphorylated hsp27 on serine residues, and two dimensional phosphopeptide mapping indicated the same sites were phosphorylated. A tumor-promoting phorbol ester, phorbol 12-myristate 13-acetate (PMA), stimulated both MAPKAP kinase-2alpha and MAPKAP kinase-2beta activity. Inhibition of MEK activation with PD 98059 or p38alpha/beta MAP kinase activity with SB203580 blocked activation by PMA. However, whereas PD 98059 inhibited only the PMA-stimulated activation, SB203580 inhibited both PMA-stimulated and basal hsp27 phosphorylation. These data demonstrate the presence of two forms of MAPKAP kinase-2 in adult ventricular myocytes. Both forms are activated indirectly by the ERK MAP kinase pathway and directly by p38 MAP kinase but independently regulated. PMID- 10821689 TI - Membrane assembly of the bacteriophage Pf3 major coat protein. AB - The Pf3 major coat protein of the Pf3 bacteriophage is stored in the inner membrane of the infected cell during the reproductive cycle. The protein consists of 44 amino acids, and contains an acidic amphipathic N-terminal domain, a hydrophobic domain, and a short basic C-terminal domain. The mainly alpha-helical membrane-bound protein traverses the membrane once, leaving the C-terminus in the cytoplasm and the N-terminus in the periplasm. A cysteine-scanning approach was followed to measure which part of the membrane-bound Pf3 protein is inside or outside the membrane. In this approach, the fluorescence probe N [(iodoacetyl)amino]ethyl-1-sulfonaphthylamine (IAEDANS) was attached to single cysteine mutants of the Pf3 coat protein. The labeled mutant coat proteins were reconstituted into the phospholipid DOPC/DOPG (80/20 molar ratio) and DOPE/DOPG (80/20 molar ratio) model membranes. We subsequently studied the fluorescence characteristics at the different positions in the protein. We measured the local polarity of the environment of the probe, as well as the accessibility of the probe to the fluorescence quencher acrylamide. The results of this study show a single membrane-spanning protein with both the C- and N-termini remaining close to the surface of the membrane. A nearly identical result was seen previously for the membrane-bound M13 coat protein. On the basis of a comparison between the results from both studies, we suggest an "L-shaped" membrane-bound model for the Pf3 coat protein. DOPE-containing model membranes revealed a higher polarity, and quenching efficiency at the membrane/water interface. Furthermore, from the outside to the inside of the membrane, a steeper polarity gradient was measured at the PE/PG interface as compared to the PC/PG interface. These results suggest a thinner interface for DOPE/DOPG than for DOPC/DOPG membranes. PMID- 10821690 TI - The sucrose permease of Escherichia coli: functional significance of cysteine residues and properties of a cysteine-less transporter. AB - The sucrose (CscB) permease belongs to the oligosaccharide:H(+) symporter family of the Major Facilitator Superfamily and is homologous to the lactose permease from Escherichia coli. Sucrose transport in cells expressing sucrose permease is completely inhibited by N-ethylmaleimide (NEM), suggesting that one or more of the seven native Cys residues may be important for transport. In this paper, each Cys residue was individually replaced with Ser, and transport activity, membrane expression, and NEM sensitivity are documented. All seven single Cys-->Ser mutants are expressed normally in the membrane and catalyze sucrose transport with activities ranging from 80% to 180% of wild type. Six of the seven Ser mutants are completely inactivated by NEM, while Cys122-->Ser permease is insensitive to the sulfhydryl reagent, indicating that NEM inhibition results from alkylation of Cys122. Subsequently, a sucrose permease devoid of Cys residues (Cys-less) was constructed in which all Cys residues were replaced with Ser simultaneously by using a series of overlap-extension PCRs. Membrane expression and kinetic parameters for Cys-less [K(m) 4.8 mM, V(max) 192 nmol min( 1) (mg of protein)(-1)] are essentially identical to those of wild type [K(m) 5.4 mM, V(max) 196 nmol min(-1) (mg of protein)(-1)]. However, Cys-less permease catalyzes sucrose accumulation to steady-state levels that are approximately 2 fold higher than those of wild type. As anticipated, Cys-less permease is completely resistant to NEM inhibition. The observations demonstrate that Cys residues play no functional role in sucrose permease. Furthermore, the approach described to create the Cys-less transporter is generally applicable to other proteins. An application of Cys-less permease in the study of the substrate binding site is presented in the accompanying paper. PMID- 10821691 TI - Functional conservation in the putative substrate binding site of the sucrose permease from Escherichia coli. AB - The sucrose (CscB) permease is the only member of the oligosaccharide:H(+) symporter family in the Major Facilitator Superfamily that transports sucrose but not lactose or other galactosides. In lactose permease (lac permease), the most studied member of the family, three residues have been shown to participate in galactoside binding: Cys148 hydrophobically interacts with the galactosyl ring, while Glu126 and Arg144 are charge paired and form H-bonds with specific galactosyl OH groups. In the present study, the role of the corresponding residues in sucrose permease, Asp126, Arg144, and Ser148, is investigated using a functional Cys-less mutant (see preceding paper). Replacement of Ser148 with Cys has no significant effect on transport activity or expression, but transport becomes highly sensitive to the sulfhydryl reagent N-ethylmaleimide (NEM) in a manner similar to that of lac permease. However, in contrast to lac permease, substrate affords no protection whatsoever against NEM inactivation of transport or alkylation with [(14)C]NEM. Neutral (Ala, Cys) mutations of Asp126 and Arg144 abolish sucrose transport, while membrane expression is not affected. Similarly, combination of two Ala mutations within the same molecule (Asp126-->Ala/Arg144- >Ala) yields normally expressed, but completely inactive permease. Conservative replacements result in highly active molecules: Asp126-->Glu permease catalyzes sucrose transport comparable to Cys-less permease, while mutant Arg144-->Lys exhibits decreased but significant activity. The observations demonstrate that charge pair Asp126-Arg144 plays an essential role in sucrose transport and suggest that the overall architecture of the substrate binding sites is conserved between sucrose and lac permeases. PMID- 10821692 TI - Recognition of base mismatches in DNA by 5,6-chrysenequinone diimine complexes of rhodium(III): a proposed mechanism for preferential binding in destabilized regions of the double helix. AB - 5,6-chrysenequinone diimine (chrysi) complexes of rhodium(III) have been shown to be versatile and specific recognition agents for mismatched base pairs in DNA. The design of these compounds was based on the hypothesis that the sterically expansive chrysi ligand, which should be too wide to readily intercalate into B DNA, would bind preferentially in the destabilized regions of the DNA helix near base mismatches. In this work, this recognition hypothesis is comprehensively explored. Comparison of the recognition patterns of the complex [Rh(bpy)(2)(chrysi)](3+) with a nonsterically demanding analogue, [Rh(bpy)(2)(phi)](3+) (phi = 9,10-phenanthrenequinone diimine), demonstrates that the chrysi ligand does indeed disfavor binding to B-DNA and generate mismatch selectivity. Examination of mismatch recognition by [Rh(bpy)(2)(chrysi)](3+) in both constant and variable sequence contexts using photocleavage assays indicates that the recognition of base mismatches is influenced by the amount that a mismatch thermodynamically destabilizes the DNA helix. Thermodynamic binding constants for the rhodium complex at a range of mismatch sites have been determined by quantitative photocleavage titration and yield values which vary from 1 x 10(6) to 20 x 10(6) M(-)(1). These mismatch-specific binding affinities correlate with independent measurements of thermodynamic destabilization, supporting the hypothesis that helix destabilization is a factor determining the binding affinity of the metal complex for the mismatched site. Although not the only factor involved in the binding of [Rh(bpy)(2)(chrysi)](3+) to mismatch sites, a model is proposed where helix destabilization acts as the "door" which permits access of the sterically demanding intercalator to the base stack. PMID- 10821693 TI - Use of duplex rigidity for stability and specificity in RNA tertiary structure. AB - The Tetrahymena group I ribozyme's oligonucleotide substrate, CCCUCUA(5), forms six base pairs with the ribozyme's internal guide sequence (IGS, 5'GGAGGG) to give the P1 duplex, and this duplex then docks into the active site via tertiary interactions. Shortening the substrate by three residues to give UCUA(5) reduces the equilibrium constant for P1 docking by approximately 200-fold even though UCUA(5) retains all the functional groups known to be involved in tertiary interactions [Narlikar, G. J., Bartley, L. E., Khosla, M., and Herschlag, D. (1999) Biochemistry 38, 14192-14204]. Here we show that the P1 duplex formed with UCUA(5) engages in all of the major tertiary interactions made by the standard P1 duplex. This suggests that the destabilization is not due to disruption of specific tertiary interactions. It therefore appears that the weaker docking of UCUA(5) arises from the increased conformational freedom of the undocked P1 duplex, which has three unpaired IGS residues and thus a larger entropic cost for docking. Further, a 2'-methoxy substitution at an IGS residue that is base-paired in the standard P1 duplex with CCCUCUA(5) but unpaired in the P1 duplex with UCUA(5) destabilizes docking of the standard P1 duplex approximately 300-fold more than it destabilizes docking of the P1 duplex formed with UCUA(5). These results suggest that fixation of groups in the context of a rigid duplex may be a general strategy used by RNA to substantially increase interaction specificity, both by aiding binding of the desired functional groups and by increasing the energetic cost of forming alternative interactions. PMID- 10821694 TI - Long-range guanine oxidation in DNA restriction fragments by a triplex-directed naphthalene diimide intercalator. AB - Naphthalene diimide (NDI), a powerful oxidant that binds avidly to DNA by intercalation, is seen to damage the 5' guanine of 5'-GG-3' sites by photoactivated charge transport through DNA. When covalently tethered to the center of a triplex-forming oligonucleotide and delivered by triplex formation within a pyrimidine.purine-pyrimidine motif to a specific site on a restriction fragment, NDI can photooxidize guanine over at least 25-38 bp in each direction from the site of binding. Charge migration occurs in both directions from the NDI intercalator and on both DNA strands of the target, but the oxidation is significantly more efficient to the 3' side of the triplex. NDI and octahedral rhodium intercalators, when tethered directly to the 5' terminus of the triplex forming strand as opposed to the center, generate significant amounts of oxidative damage only in the immediate vicinity of the intercalation site. Given that long-range charge transport depends on DNA stacking, these results suggest that the base stack is distorted at the 5' end of the triplex region in the duplex-triplex junction. Targeting of photooxidative damage by triplex formation extends our previous studies of long-range charge transport to significantly longer DNA sequences through a strategy that does not require covalent attachment of the photooxidant to the DNA being probed. Moreover, triplex targeting of oxidative damage provides for the first time a typical distance distribution for genomic charge transport of approximately 200 A around the oxidant. PMID- 10821695 TI - Functional analyses of two cellular binding domains of bovine lactadherin. AB - The glycoprotein bovine lactadherin (formerly known as PAS-6/7) comprises two EGF like domains and two C-like domains found in blood clotting factors V and VIII. Bovine lactadherin binds to alpha(v)beta(5) integrin in an RGD-dependent manner and also to phospholipids, especially phosphatidyl serine. To define and characterize these bindings the interactions between lactadherin and different mammalian cell types were investigated. Using recombinant forms of bovine lactadherin, the human breast carcinomas MCF-7 cells expressing the alpha(v)beta(5) integrin receptor were shown to bind specifically to RGD containing lactadherin but not to a mutated RGE lactadherin. A monoclonal antibody against the alpha(v)beta(5) integrin receptor and a synthetic RGD containing peptide inhibited the adhesion of MCF-7 cells to lactadherin. Green monkey kidney MA-104 cells, also expressing the alpha(v)beta(3) together with the alpha(v)beta(5) integrin, showed binding to bovine lactadherin via both integrins. To investigate the interaction of lipid with lactadherin two fragments were expressed corresponding to the C1C2 domains and the C2 domain. Both fragments bound to phosphatidyl serine in a concentration-dependent manner with an affinity similar to native lactadherin (K(d) = 1.8 nM). A peptide corresponding to the C-terminal part of the C2 domain inhibited the binding of lactadherin to phospholipid in a concentration-dependent manner, and finally it was shown that lactadherin mediates binding between artificial phosphatidyl serine membranes and MCF-7 cells. Taken together these results show that lactadherin can act as link between two surfaces by binding to integrin receptors through its N-terminus and to phospholipids through its C-terminus. PMID- 10821696 TI - Selection of viral RNA-derived tRNA-like structures with improved valylation activities. AB - The tRNA-like structure (TLS) of turnip yellow mosaic virus (TYMV) RNA was previously shown to be efficiently charged by yeast valyl-tRNA synthetase (ValRS). This RNA has a noncanonical structure at its 3'-terminus but mimics a tRNA L-shaped fold, including an anticodon loop containing the major identity nucleotides for valylation, and a pseudoknotted amino acid accepting domain. Here we describe an in vitro selection experiment aimed (i) to verify the completeness of the valine identity set, (ii) to elucidate the impact of the pseudoknot on valylation, and (iii) to investigate whether functional communication exists between the two distal anticodon and amino acid accepting domains. Valylatable variants were selected from a pool of 2 x 10(13) RNA molecules derived from the TYMV TLS randomized in the anticodon loop nucleotides and in the length (1-6 nucleotides) and sequence of the pseudoknot loop L1. After nine rounds of selection by aminoacylation, 42 have been isolated. Among them, 17 RNAs could be efficiently charged by yeast ValRS. Their sequence revealed strong conservation of the second and the third anticodon triplet positions (A(56), C(55)) and the very 3'-end loop nucleotide C(53). A large variability of the other nucleotides of the loop was observed and no wild-type sequence was recovered. The selected molecules presented pseudoknot domains with loop L1 varying in size from 3-6 nucleotides and some sequence conservation, but did neither reveal the wild-type combination. All selected variants are 5-50 times more efficiently valylated than the wild-type TLS, suggesting that the natural viral sequence has emerged from a combination of evolutionary pressures among which aminoacylation was not predominant. This is in line with the role of the TLS in viral replication. PMID- 10821698 TI - Fluorescence quenching analysis of the association and dissociation of a diarylheterocycle to cyclooxygenase-1 and cyclooxygenase-2: dynamic basis of cyclooxygenase-2 selectivity. AB - Cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) are the enzymes responsible for the biosynthesis of the precursor to the biologically active prostaglandins, prostacyclin, and thromboxane and are the molecular targets for nonsteroidal antiinflammatory drugs (NSAIDs). Selective COX-2 inhibitors are antiinflammatory and analgesic but lack gastrointestinal toxicity, an undesirable side effect attributed to COX-1 inhibition. Crystallographic analysis of selective COX inhibitors complexed with either isoform provides some information about the molecular determinants of selectivity but does not provide information about the dynamics of inhibitor association/dissociation. We employed rapid-mixing techniques and fluorescence quenching to monitor the association and dissociation of a selective COX-2 inhibitor to COX-1 or COX-2. The association of the fluorescent diaryloxazole, SC299, with both enzymes occurs in a time-dependent fashion. Its binding to COX-2 occurs in three kinetically distinct steps whereas its binding to COX-1 occurs in two steps. In contrast to the relatively rapid association of SC299 with both enzymes, its dissociation from COX-2 is quite slow and occurs over several hours whereas the dissociation from COX-1 is complete in less than 1 min. The selectivity of SC299 as a COX-2 inhibitor correlates to its relative rates of dissociation from the two COX isoforms. A model is proposed for diarylheterocycle binding to COX's that integrates these kinetic data with available structural information. PMID- 10821697 TI - Role of beta Arg211 in the active site of human beta-hexosaminidase B. AB - Tay-Sachs or Sandhoff disease results from a deficiency of either the alpha- or the beta-subunits of beta-hexosaminidase A, respectively. These evolutionarily related subunits have been grouped with the "Family 20" glycosidases. Molecular modeling of human hexosaminidase has been carried out on the basis of the three dimensional structure of a bacterial member of Family 20, Serratia marcescens chitobiase. The primary sequence identity between the two enzymes is only 26% and restricted to their active site regions; therefore, the validity of this model must be determined experimentally. Because human hexosaminidase cannot be functionally expressed in bacteria, characterization of mutagenized hexosaminidase must be carried out using eukaryotic cell expression systems that all produce endogenous hexosaminidase activity. Even small amounts of endogenous enzyme can interfere with accurate K(m) or V(max) determinations. We report the expression, purification, and characterization of a C-terminal His(6)-tag precursor form of hexosaminidase B that is 99.99% free of endogenous enzyme from the host cells. Control experiments are reported confirming that the kinetic parameters of the His(6)-tag precursor are the same as the untagged precursor, which in turn are identical to the mature isoenzyme. Using highly purified wild type and Arg(211)Lys-substituted hexosaminidase B, we reexamine the role of Arg(211) in the active site. As we previously reported, this very conservative substitution nevertheless reduces k(cat) by 500-fold. However, the removal of all endogenous activity has now allowed us to detect a 10-fold increase in K(m) that was not apparent in our previous study. That this increase in K(m) reflects a decrease in the strength of substrate binding was confirmed by the inability of the mutant isozyme to efficiently bind an immobilized substrate analogue, i.e., a hexosaminidase affinity column. Thus, Arg(211) is involved in substrate binding, as predicted by the chitobiase model, as well as catalysis. PMID- 10821700 TI - Structural requirements for the interdomain linker of alpha subunit of Escherichia coli RNA polymerase. AB - The carboxy-terminal domain of the alpha subunit of Escherichia coli RNA polymerase, which is connected with the core part of RNA polymerase through a long flexible linker, plays decisive roles in transcription activation by directly interacting with a large number of transcription factors and upstream (UP) element DNA. Here we constructed a set of mutant RNA polymerases, each containing a mutant alpha subunit with an altered interdomain linker. Deletion of three amino acids from the linker exhibited 50% inhibition of cAMP receptor protein- (CRP-) dependent lac P1 transcription. Deletion of six amino acids completely knocked out the activity. Insertion of three amino acids did not affect the activity, whereas 40-60% inhibition was observed after insertion of one, two, or four amino acids. Substitution of 10 consecutive glycine residues resulted in nearly 90% reduction of the CRP-dependent activity, whereas 50% activity was retained after substitution of 10 proline residues or a sequence expected to form a strong alpha-helix. Essentially the same results were obtained with UP element-dependent rrnB P1 transcription. These observations altogether suggest that (i) sufficient length of the interdomain linker is required for transcription activation mediated by the alpha carboxy-terminal domain, (ii) the linker is not totally unstructured but has structural and torsional preferences to facilitate positioning of the carboxy-terminal domain to a proper location for the interaction with CRP and UP element, and (iii) CRP-dependent activation and UP element-dependent activation share a common intermediary state in which the positioning of the alpha carboxy-terminal domain is of primary importance. PMID- 10821699 TI - Internal loop/bulge and hairpin loop of the iron-responsive element of ferritin mRNA contribute to maximal iron regulatory protein 2 binding and translational regulation in the iso-iron-responsive element/iso-iron regulatory protein family. AB - Iron-responsive elements (IREs), a natural group of mRNA-specific sequences, bind iron regulatory proteins (IRPs) differentially and fold into hairpins [with a hexaloop (HL) CAGUGX] with helical distortions: an internal loop/bulge (IL/B) (UGC/C) or C-bulge. C-bulge iso-IREs bind IRP2 more poorly, as oligomers (n = 28 30), and have a weaker signal response in vivo. Two trans-loop GC base pairs occur in the ferritin IRE (IL/B and HL) but only one in C-bulge iso-IREs (HL); metal ions and protons perturb the IL/B [Gdaniec et al. (1998) Biochemistry 37, 1505-1512]. IRE function (translation) and physical properties (T(m) and accessibility to nucleases) are now compared for IL/B and C-bulge IREs and for HL mutants. Conversion of the IL/B into a C-bulge by a single deletion in the IL/B or by substituting the HL CG base pair with UA both derepressed ferritin synthesis 4-fold in rabbit reticulocyte lysates (IRP1 + IRP2), confirming differences in IRP2 binding observed for the oligomers. Since the engineered C bulge IRE was more helical near the IL/B [Cu(phen)(2) resistant] and more stable (T(m) increased) and the HL mutant was less helical near the IL/B (ribonuclease T1 sensitive) and less stable (T(m) decreased), both CG trans-loop base pairs contribute to maximum IRP2 binding and translational regulation. The (1)H NMR spectrum of the Mg-IRE complex revealed, in contrast to the localized IL/B effects of Co(III) hexaammine observed previously, perturbation of the IL/B plus HL and interloop helix. The lower stability and greater helix distortion in the ferritin IL/B-IRE compared to the C-bulge iso-IREs create a combinatorial set of RNA/protein interactions that control protein synthesis rates with a range of signal sensitivities. PMID- 10821701 TI - Identification of cis-ethanesemidione as the organic radical derived from glycolaldehyde in the suicide inactivation of dioldehydrase and of ethanolamine ammonia-lyase. AB - The hydrate of glycolaldehyde is a substrate analogue that induces the formation of cob(II)alamin and 5'-deoxyadenosine from adenosylcobalamin at the active site of dioldehydrase, and the resulting complex is inactive. The carbon atoms of glycolaldehyde hydrate remain bound to this complex, and it has been postulated that the first step or steps of the catalytic process on glycolaldehyde hydrate generate an intermediate that undergoes a destructive side reaction leading to inactivation of the enzyme [Wagner, O. W., Lee, H. A., Jr., Frey, P. A., and Abeles, R. H. (1966) J. Biol. Chem. 249, 1751-1762]. All evidence suggests that dioldehydrase reaction proceeds by a radical mechanism, and the glycolaldehyde hydrate is expected to be converted initially into a radical. Electron paramagnetic resonance (EPR) spectroscopic analysis of the inactivated complex shows that glycolaldehyde is transformed into a cis-ethanesemidione radical that is weakly spin-coupled to the cob(II)alamin in the active site of the enzyme. This radical has been identified by analysis of EPR spectra obtained from samples with (13)C- and (2)H-labeled forms of glycolaldehyde. The analysis shows that the stable radical associated with the inactive complex is symmetrical and that it contains a single solvent-exchangeable proton, consistent with a cis ethanesemidione. Glycolaldehyde also inactivates ethanolamine ammonia-lyase (EAL). EPR studies of ethanolamine ammonia-lyase reveal that treatment with glycolaldehyde also results in formation of an ethanesemidione radical bound in the active site. The suicide inactivation in both enzymatic reactions is postulated to result from formation of this stable radical, which cannot react further to abstract a hydrogen atom from 5'-deoxyadenosine. Analysis of the electron spin-spin coupling between the semidione radicals and cob(II)alamin in both enzymes indicates that the distance between the radical and Co(2+) is approximately 11 A in each case. PMID- 10821702 TI - Catalytic reaction pathway for the mitogen-activated protein kinase ERK2. AB - The structural, functional, and regulatory properties of the mitogen-activated protein kinases (MAP kinases) have long attracted considerable attention owing to the critical role that these enzymes play in signal transduction. While several MAP kinase X-ray crystal structures currently exist, there is by comparison little mechanistic information available to correlate the structural data with the known biochemical properties of these molecules. We have employed steady state kinetic and solvent viscosometric techniques to characterize the catalytic reaction pathway of the MAP kinase ERK2 with respect to the phosphorylation of a protein substrate, myelin basic protein (MBP), and a synthetic peptide substrate, ERKtide. A minor viscosity effect on k(cat) with respect to the phosphorylation of MBP was observed (k(cat) = 10 +/- 2 s(-1), k(cat)(eta) = 0.18 +/- 0.05), indicating that substrate processing occurs via slow phosphoryl group transfer (12 +/- 4 s(-1)) followed by the faster release of products (56 +/- 4 s(-1)). At an MBP concentration extrapolated to infinity, no significant viscosity effect on k(cat)/K(m(ATP)) was observed (k(cat)/K(m(ATP)) = 0.2 +/- 0.1 microM(-1) s(-1), k(cat)/K(m(ATP))(eta) = -0.08 +/- 0.04), consistent with rapid-equilibrium binding of the nucleotide. In contrast, at saturating ATP, a full viscosity effect on k(cat)/K(m) for MBP was apparent (k(cat)/K(m(MBP)) = 2.4 +/- 1 microM( 1) s(-1), k(cat)/K(m(MBP))(eta) = 1.0 +/- 0.1), while no viscosity effect was observed on k(cat)/K(m) for the phosphorylation of ERKtide (k(cat)/K(m(ERKtide)) = (4 +/- 2) x 10(-3) microM(-1) s(-1), k(cat)/K(m(ERKtide))(eta) = -0.02 +/- 0.02). This is consistent with the diffusion-limited binding of MBP, in contrast to the rapid-equilibrium binding of ERKtide, to form the ternary Michaelis complex. Calculated values for binding constants show that the estimated value for K(d(MBP)) (/= 1.5 mM). The dramatically higher catalytic efficiency of MBP in comparison to that of ERKtide ( approximately 600-fold difference) is largely attributable to the slow dissociation rate of MBP (/=56 s(-1)), from the ERK2 active site. PMID- 10821703 TI - Application of similarity matrices and genetic neural networks in quantitative structure-activity relationships of 2- or 4-(4-Methylpiperazino)pyrimidines: 5 HT(2A) receptor antagonists. AB - Antagonists of the 5-HT(2A) receptor are being used to treat many psychiatric disorders. The present work focuses on a group of 27 antagonists possessing varying affinities toward the receptor. These are 26 title compounds and clozapine as a reference antagonist. The active conformers of the conformationally flexible ligands were proposed by using the active rigid analogue approach and performing similarity calculations. The calculations involved genetic neural network (GNN) computations deriving QSARs from similarity matrices (SM) with cross-validated correlation coefficients exceeding 0.92. The performance of neural networks with variety of architectures was studied. As the computations were performed for cations and neutral molecules separately, the relevance of the ligand charging is discussed. PMID- 10821704 TI - Design and synthesis of Cyclopenta[g]quinazoline-based antifolates as inhibitors of thymidylate synthase and potential antitumor agents(,). AB - Following the development of raltitrexed, the synthesis of nonpolyglutamatable inhibitors of TS that do not use the reduced folate carrier (RFC) for cellular entry should provide compounds which overcome mechanisms of resistance to folate based inhibitors of TS that are associated with decreased/altered folylpolyglutamate synthetase (FPGS) expression and/or an impaired RFC. Examination of a computer graphics model of the humanized Escherichia coli TS enzyme with quinazoline inhibitors of TS, such as 1 bound in the active site of the enzyme, suggested that conformational restriction introduced by bridging the C9 with C7 to form a pentacycle may be beneficial for binding to TS. That led to the synthesis of a series of potent cyclopenta[g]quinazoline-based inhibitors of the enzyme in which the glutamyl residue associated with classical antifolates was replaced with a variety of glutamate-derived ligands; the most potent inhibitor being the L-Glu-gamma-D-GluT(alpha) derivative 7j. In the mouse L1210:1565 cell line (mutant RFC), the majority of these compounds had activity equal or only slightly greater compared with the parental L1210 cell line, indicating a reduced dependence on the RFC for cellular uptake in the L1210 cell line. PMID- 10821705 TI - Synthesis and evaluation of "AZT-HEPT", "AZT-pyridinone", and "ddC-HEPT" conjugates as inhibitors of HIV reverse transcriptase. AB - To test the concept that HIV reverse transcriptase could be effectively inhibited by "mixed site inhibitors", a series of seven conjugates containing both a nucleoside analogue component (AZT 1, ddC 2) and a nonnucleoside type inhibitor (HEPT analogue 12, pyridinone 27) were synthesized and evaluated for their ability to block HIV replication. The (N-3 and C-5)AZT-HEPT conjugates 15, 22, and 23 displayed 2-5 microM anti-HIV activity, but they had no effect on the replication of HIV-2 or the HIV-1 strain with the Y181C mutation. The (C-5)AZT pyridinone conjugates 34-37 were found to be inactive. In marked contrast, the ddC-HEPT molecule 26 displayed the same potency (EC(50) = 0.45 microM) against HIV-1 (wild type and the Y181C nevirapine-resistant strain) and HIV-2 in cell culture. No synergistic effect was observed for these bis-substrate inhibitors, suggesting that the two individual inhibitor components in these molecules do not bind simultaneously in their respective sites. Interestingly, however, the results indicate that the AZT-HEPT conjugates and the ddC-HEPT derivative 26 inhibit reverse transcriptase (RT) in an opposite manner. One explanation for this difference is that the former compounds interact preferentially with the hydrophobic pocket in RT, whereas 26 (after supposed triphosphorylation) inhibits RT through binding in the catalytic site. PMID- 10821706 TI - 1,3,5-Trialkyl-2,4,6-triiodobenzenes: novel X-ray contrast agents for gastrointestinal imaging. AB - Examination of the gastrointestinal (GI) tract has been performed for decades using barium sulfate. Although this agent has many recognized limitations including extreme radiopacity, poor intrinsic affinity for the GI mucosa, and very high density, no alternative contrast agents have emerged which produce comparable or better contrast visualization. In fact, the various techniques of the GI radiologic examination (i.e., single contrast, double contrast, biphasic) were developed to compensate for its limitations. Each of these techniques requires complex patient manipulation to achieve adequate mucosal coating or compression to overcome the marked radiopacity of barium sulfate in order to obtain a diagnostically useful examination. A series of novel radiopaque oils, the 1,3, 5-trialkyl-2,4,6-triiodobenzenes, was designed to improve the efficacy, stability, and safety of barium formulations. These substances were prepared in two steps from 1,3,5-trichlorobenzene. Compound 17 (1,3,5-tri-n-hexyl-2,4,6 triiodobenzene), formulated as an oil-in-water emulsion, was found to be well tolerated in rodents (mice, hamsters, rats) following acute oral and/or intraperitoneal administrations at 4 times the anticipated human clinical dose. No metabolism of 17 was detected in rat, hamster, dog, monkey, or human hepatic microsomes, suggesting the lack of oral toxicity was a consequence of poor absorption. In imaging experiments in dogs, emulsions of 17 have demonstrated excellent mucosal coating and improved radiodensity relative to barium sulfate suspensions. On the basis of the preliminary imaging and toxicity data, compound 17 was selected as a potential development candidate. PMID- 10821707 TI - Modeling of the inhibition of retroviral integrases by styrylquinoline derivatives. AB - Styrylquinoline derivatives, known to be potent inhibitors of HIV-1 integrase, have been experimentally tested for their inhibitory effect on the disintegration reaction catalyzed by catalytic cores of HIV-1 and Rous sarcoma virus (RSV) integrases. A modified docking protocol, consisting of coupling a grid search method with full energy minimization, has been specially designed to study the interaction between the inhibitors and the integrases. The inhibitors consist of two moieties that have hydroxyl and/or carboxyl substituents: the first moiety is either benzene, phenol, catechol, resorcinol, or salicycilic acid; the hydroxyl substituents on the second (quinoline) moiety may be in the keto or in the enol forms. Several tautomeric forms of the drugs have been docked to the crystallographic structure of the RSV catalytic core. The computed binding energy of the keto forms correlates best with the measured inhibitory effect. The docking procedure shows that the inhibitors bind closely to the crystallographic catalytic Mg(2+) dication. Additional quantum chemistry computations show that there is no direct correlation between the binding energy of the drugs with the Mg(2+) dication and their in vitro inhibitory effect. The designed method is a leading way for identification of potent integrase inhibitors using in silico experiments. PMID- 10821708 TI - 4-Alkyl- and 4-cinnamylglutamic acid analogues are potent GluR5 kainate receptor agonists. AB - Enantiomerically pure (2S,4R)-4-substituted glutamic acids were prepared and tested for homomeric GluR5 and GluR6 kainate subtype receptor affinity. Some of the 4-cinnamyl analogues showed high selectivity and potency (K(i) < 25 nM) for the GluR5 receptors. The greatest selectivity and potency were achieved with the 3-(2-naphthyl)prop-2-enyl compound. This compound, LY339434, has negligible activity at the AMPA and kainate receptors GluR1, -2, -4 and -6. Although, LY339434 shows agonist activity at NMDA receptors in cultural hippocampal neurons (approximate EC(50) of 2.5 microM), we consider that LY339434 should be a useful pharmacological tool for the investigation of the functional role of GluR5 kainate receptors. PMID- 10821709 TI - Design, synthesis, and preliminary pharmacological evaluation of 1, 4 diazabicyclo[4.3.0]nonan-9-ones as a new class of highly potent nootropic agents. AB - Several 4-substituted 1,4-diazabicyclo[4.3.0]nonan-9-ones have been synthesized and tested in vivo on mouse passive avoidance test, to evaluate their nootropic activity. The results show that they represent a new class of nootropic drugs with a pharmacological profile very similar to that of piracetam, showing much higher potency with respect to the reference. Among the compounds studied, 7 (DM 232) shows outstanding potency, being active at the dose of 0. 001 mg kg(-1) sc. PMID- 10821710 TI - A novel approach for the virtual screening and rational design of anticancer compounds. AB - A topological substructural approach to molecular design (TOSS-MODE) has been introduced for the selection and design of anticancer compounds. A quantitative model that discriminates anticancer compounds from the inactive ones in a training series was obtained. This model permits the correct classification of 91.43% of compounds in an external prediction set with only 1.43% of false actives and 7. 14% of false inactives. The model developed is then used in a simulation of a virtual search for Ras FTase inhibitors; 87% of the Ras FTase inhibitors used in this simulated search were correctly classified, thus indicating the ability of the TOSS-MODE model of finding lead compounds with novel structures and mechanism of action. Finally, a series of carbonucleosides was designed, and the compounds were classified as active/inactive anticancer compounds by using the model developed here. From the compounds so-designed, 20 were synthesized and evaluated experimentally for their antitumor effects on the proliferation of murine leukemia cells (L1210/0) and human T-lymphocyte cells (Molt4/C8 and CEM/0); 80% of these compounds were well-classified, as active or inactive, and only two pairs of isomeric compounds were false actives. The chloropurine derivatives were the most active compounds, especially compounds 6c, d. PMID- 10821712 TI - Quantized surface complementarity diversity (QSCD): a model based on small molecule-target complementarity. AB - A model of molecular diversity is presented. The model, termed "Quantized Surface Complementarity Diversity" (QSCD), defines molecular diversity by measuring molecular complementarity to a fully enumerated set of theoretical target surfaces. Molecular diversity space is defined as the molecular complement to this set of enumerated surfaces. Using a set of known test compounds, the model is shown to be biologically relevant, consistently scoring known actives as similar. At the resolution of the model, which examines molecules "quantized" into 4.24 A cubic units and treats four points of specific energetic complementarity, the minimum number of compounds needed to fully cover molecular diversity space up to volume 1070 cubic A is estimated to be on the order of 24 million molecules. Most importantly, QSCD allows for individual points in diversity space to be filled by direct modeling of molecular libraries into detailed 3D templates of shape and functionality. PMID- 10821711 TI - Biosensor analysis of the interaction between immobilized human serum albumin and drug compounds for prediction of human serum albumin binding levels. AB - The interactions between a set of drugs, selected on the basis of reported human serum albumin (HSA) binding levels, and immobilized HSA were investigated using surface plasmon resonance technology. Major HSA binding sites were available after immobilization. The intensity of the signal obtained from the interaction of the drug with the HSA surface was correlated with the reported HSA binding level. Drugs were classified into groups corresponding to high, medium, or low HSA binding based on the injection of the drug at 80 microM concentration. A set of 10 drugs binding to alpha(1)-acid glycoprotein (AGP) was also investigated and correlated with reported AGP binding data. The throughput of the presented assay is 100 compounds/24 h, and the sample consumption is less than 100 microL (8 nmol). PMID- 10821713 TI - SAR of 9-amino-1,2,3,4-tetrahydroacridine-based acetylcholinesterase inhibitors: synthesis, enzyme inhibitory activity, QSAR, and structure-based CoMFA of tacrine analogues. AB - In this study, we attempted to derive a comprehensive SAR picture for the class of acetylcholinesterase (AChE) inhibitors related to tacrine, a drug currently in use for the treatment of the Alzheimer's disease. To this aim, we synthesized and tested a series of 9-amino-1,2,3,4-tetrahydroacridine derivatives substituted in the positions 6 and 7 of the acridine nucleus and bearing selected groups on the 9-amino function. By means of the Hansch approach, QSAR equations were obtained, quantitatively accounting for both the detrimental steric effect of substituents in position 7 and the favorable electron-attracting effect exerted by substituents in positions 6 and 7 of the 9-amino-1,2,3,4-tetrahydroacridine derivatives. The three-dimensional (3D) properties of the inhibitors were taken into consideration by performing a CoMFA analysis on the series of AChE inhibitors made by 12 9-amino-1,2,3, 4-tetrahydroacridines and 13 11H-indeno[1,2 b]quinolin-10-ylamines previously developed in our laboratory. The alignment of the molecules to be submitted to the CoMFA procedure was carried out by taking advantage of docking models calculated for the interactions of both the unsubstituted 9-amino-1,2,3,4-tetrahydroacridine and 11H-indeno[1,2-b]quinolin-10 ylamine with the target enzyme. A highly significant CoMFA model was obtained using the steric field alone, and the features of such a 3D QSAR model were compared with the classical QSAR equations previously calculated. The two models appeared consistent, the main aspects they had in common being (a) the individuation of the strongly negative contribution of the substituents in position 7 of tacrine and (b) a tentative assignment of the hydrophobic character to the favorable effect exerted by the substituents in position 6. Finally, a new previously unreported tacrine derivative designed on the basis of both the classical and the 3D QSAR equations was synthesized and kinetically evaluated, to test the predictive ability of the QSAR models. The 6-bromo-9-amino-1,2,3,4 tetrahydroacridine was predicted to have a pIC(50) value of 7.31 by the classical QSAR model and 7.40 by the CoMFA model, while its experimental IC(50) value was equal to 0.066 (+/-0.009) microM, corresponding to a pIC(50) of 7.18, showing a reasonable agreement between predicted and observed AChE inhibition data. PMID- 10821714 TI - Inhibition of clinically relevant mutant variants of HIV-1 by quinazolinone non nucleoside reverse transcriptase inhibitors. AB - A series of 4-alkenyl and 4-alkynyl-3, 4-dihydro-4-(trifluoromethyl)-2-(1H) quinazolinones were found to be potent non-nucleoside reverse transcriptase inhibitors (NNRTIs) of human immunodeficiency virus type-1 (HIV-1). The 4-alkenyl 3, 4-dihydro-4-(trifluoromethyl)-2-(1H)-quinazolinones DPC 082 and DPC 083 and the 4-alkynyl-3, 4-dihydro-4-(trifluoromethyl)-2-(1H)-quinazolinones DPC 961 and DPC 963 were found to exhibit low nanomolar potency toward wild-type RF virus (IC(90) = 2.0, 2.1, 2.0, and 1.3 nM, respectively) and various single and many multiple amino acid substituted HIV-1 mutant viruses. The increased potency is combined with favorable plasma serum protein binding as demonstrated by improvements in the percent free drug in human plasma when compared to efavirenz: 3.0%, 2.0%, 1.5%, 2. 8%, and 0.2-0.5% for DPC 082, DPC 083, DPC 961, DPC 963, and efavirenz, respectively. PMID- 10821715 TI - A new class of HIV-1 integrase inhibitors: the 3,3,3', 3'-tetramethyl-1,1' spirobi(indan)-5,5',6,6'-tetrol family. AB - Integration is a required step in HIV replication, but as yet no inhibitors of the integration step have been developed for clinical use. Many inhibitors have been identified that are active against purified viral-encoded integrase protein; of these, many contain a catechol moiety. Though this substructure contributes potency in inhibitors, it is associated with toxicity and so the utility of catechol-containing inhibitors has been questioned. We have synthesized and tested a systematic series of derivatives of a catechol-containing inhibitor (1) with the goal of identifying catechol isosteres that support inhibition. We find that different patterns of substitution on the aromatic ring suffice for inhibition when Mn(2+) is used as a cofactor. Importantly, the efficiency is different when Mg(2+), the more likely in vivo cofactor, is used. These data emphasize the importance of assays with Mg(2+) and offer new catechol isosteres for use in integrase inhibitors. PMID- 10821716 TI - Novel antiarthritic agents with 1,2-isothiazolidine-1,1-dioxide (gamma-sultam) skeleton: cytokine suppressive dual inhibitors of cyclooxygenase-2 and 5 lipoxygenase. AB - Various 1,2-isothiazolidine-1,1-dioxide (gamma-sultam) derivatives containing an antioxidant moiety, 2,6-di-tert-butylphenol substituent, were prepared. Some compounds, which have a lower alkyl group at the 2-position of the gamma-sultam skeleton, showed potent inhibitory effects on both cyclooxygenase (COX)-2 and 5 lipoxygenase (5-LO), as well as production of interleukin (IL)-1 in in vitro assays. They also proved to be effective in several animal arthritic models without any ulcerogenic activities. Among these compounds, (E)-(5)-(3,5-di-tert butyl-4-hydroxybenzylidene)-2-ethyl-1, 2-isothiazolidine-1,1-dioxide (S-2474) was selected as an antiarthritic drug candidate and is now under clinical trials. The structure-activity relationships (SAR) examined and some pharmacological evaluations are described. PMID- 10821717 TI - Discovery of novel, potent, and selective small-molecule CCR5 antagonists as anti HIV-1 agents: synthesis and biological evaluation of anilide derivatives with a quaternary ammonium moiety. AB - The search for new small-molecule CCR5 antagonists by high-throughput screening (HTS) of the Takeda chemical library using [(125)I]RANTES and CHO/CCR5 cells led to the discovery of lead compounds (A, B) with a quaternary ammonium or phosphonium moiety, which were synthesized to investigate new MCP-1 receptor antagonists. A series of novel anilide derivatives 1 with a quaternary ammonium moiety were designed, synthesized, and tested for their CCR5 antagonistic activity. Through the optimization of lead compounds, we have found N,N-dimethyl N-[4-[[[2-(4-methylphenyl)-6, 7-dihydro-5H-benzocyclohepten-8 yl]carbonyl]amino]benzyl]tetrahydr o-2 H-pyran-4-aminium chloride (1r, TAK-779) as a highly potent and selective nonpeptide CCR5 antagonist with a IC(50) value of 1.4 nM in the binding assay. Compound 1r also inhibited the replication of macrophage (M)-tropic HIV-1 (Ba-L strain) in both MAGI-CCR5 cells and PBMCs with EC(50) values of 1.2 and 3.7 nM, respectively. The synthesis and structure activity relationships of 1r and its related compounds are detailed. PMID- 10821718 TI - Novel conformationally constrained tropane analogues by 6-endo-trig radical cyclization and stille coupling - switch of activity toward the serotonin and/or norepinephrine transporter. AB - A novel class of tricyclic tropane analogues has been synthesized by making use of radical cyclization technology in combination with the Stille coupling reaction. As hybrids between tropanes and quinuclidines, these tropaquinuclidines represent a significant structural departure from many of the other classes of tropane ligands synthesized to date. This structure class is characterized by the boat conformation of the tropane ring and the orientation of the additional bridge (and therefore of the nitrogen lone pair) together with the unusual placement of the aromatic moiety. All compounds were tested for their ability to inhibit monoamine reuptake under identical conditions. The ability to inhibit reuptake of dopamine in comparison to cocaine is generally decreased in this series but for one compound. (1S,3R, 6S)-(Z)-9-(thienylmethylene)-7 azatricyclo[4.3.1.0(3, 7)]decane-2beta-carboxylic acid methyl ester (5h) exhibits reasonable activity at the dopamine transporter (DAT) (K(i) = 268 nM) and good activity at the norepinephrine transporter (NET) (K(i) = 26 nM). The potency and selectivity shown by some of these ligands for the NET, serotonine transporter (SERT), or NET/SERT is striking, particularly in view of the displacement of the aromatic ring in this series from its usual position at C-3 in the WIN analogues. Thus, (1S,3R,6S)-(Z)-9-(4-biphenylylmethylene)-7-azatricyclo[4.3.1 . 0(3,7)]decane-2beta-carboxylic acid methyl ester (5a) is a selective inhibitor of norepinephrine reuptake (K(i) = 12 nM). Its p-methoxy analogue 5c is a mixed inhibitor of norepinephrine and serotonin reuptake (K(i) = 187 nM at the NET and 56 nM at the SERT). The most active and selective compound we found in the present series is compound 8b [(1S,3R,6S)-2-(acetoxymethyl)-(Z)-9-(3, 4 dichlorophenylmethylene)-7-azatricyclo[4.3.1.0(3,7)]decane ]. This compound is a potent (K(i) = 1.6 nM) and selective inhibitor of serotonin reuptake into rat midbrain synaptosomes. Its selectivity is about 400-fold over the NET and about 1000-fold over the DAT. The results of this study further demonstrate the possibility of tuning the selectivity of tropane analogues toward the SERT or NET binding site. The ligands disclosed herein provide additional pharmacological tools of use in attempting to correlate structure and transporter selectivity with in vivo studies of behavioral outcomes. PMID- 10821719 TI - Structure-activity study on the in vitro antiprotozoal activity of glutathione derivatives. AB - A series of N-, S-, and COOH-blocked glutathione derivatives were evaluated against the pathogenic parasites Trypanosoma brucei, Trypanosoma cruzi, and Leishmania donovani in vitro, to identify the determinants necessary for activity and for further development into an active lead structure. The results show that N,S-blocked glutathione diesters are the most effective inhibitors of T. brucei with structures 14-16 being the most active, 14 having an IC(50) approximately 1.9 microM. The toxicity effects observed for glutathione derivatives 12, 14, and 16 have been correlated to the K562 antileukemic activity of these compounds and their inhibitory effects on the glyoxalase system of the host. Diester compounds based on S-2,4-dinitrophenylglutathione (17-22) were found to be significantly better inhibitors of T. brucei with ED(50)'s in the range 16-0.19 microM. Compounds 19 and 20 were the two best inhibitors, with an ED(50) of approximately 1.07 and 0.19 microM, respectively; however 20 displayed toxicity in parasitic assays. Monoesters, monoamides, and diamides tested generally exhibited low in vitro activity. The compounds did not inhibit glutathionylspermidine synthetase and trypanothione reductase enzyme targets in the unique trypanothione pathway of these parasites. Diester compounds per se were considered to be ineffective inhibitors of trypanothione metabolism suggesting that these compounds might act as prodrugs, being hydrolyzed in situ into a variety of glutathione derivatives which include combinations of monoesters, free acids, and amines, some of which are inhibitors of trypanothione metabolism. PMID- 10821720 TI - 7-Methyl-6,7,8,9,14,15-hexahydro-5H-benz[d]indolo[2,3-g]azecine: a new heterocyclic system and a new lead compound for dopamine receptor antagonists. AB - Partially hydrogenated derivatives of the new heterocyclic ring systems benz[d]indolo[2,3-g]azecine and bisindolo[3,2-d][2, 3-g]azecine were synthesized starting from lactones and amines via the described synthetic methods. In binding assays with rat striatal receptors, 7-methyl-6,7,8,9,14,15-hexahydro-5H benz[d]indolo[2, 3-g]azecine (LE 300) proved to be of high affinity for the D(1) binding site (K(i) = 0.08 nmol for displacement of [(3)H]SCH23390), being superior in this assay to standards such as butaclamol and SCH23390. This compound was characterized as a dopamine antagonist by conditioned avoidance response test with mice. Thus, LE 300 represents the lead of a new class of dopamine antagonists for future investigations. PMID- 10821721 TI - Dose-responses over time to inhaled fluticasone propionate treatment of exercise- and methacholine-induced bronchoconstriction in children with asthma. AB - When treating bronchial hyperresponsiveness to so-called direct and indirect stimuli, distinct pathophysiological mechanisms might require differences in dose and duration of inhaled corticosteroid therapy. To test this hypothesis in children with asthma, we investigated the time- and dose-dependent effects of 2 doses of fluticasone propionate (FP, 100 or 250 microg bid.) in improving exercise- (EIB) and methacholine-induced bronchoconstriction during 6 months of treatment, using a placebo-controlled parallel group study design. Thirty-seven children with asthma (aged 6 to 14 years; forced expired volume in 1 sec (FEV(1)) >/=70% predicted; EIB >/=20% fall in FEV(1) from baseline; no inhaled steroids during the past 4 months) participated in a double-blind, placebo-controlled, 3 arm parallel study. Children receiving placebo were re-randomized to active treatment after 6 weeks. Standardized dry air treadmill exercise testing (EIB expressed as %fall in FEV(1) from baseline) and methacholine challenge using a dosimetric technique (expressed as PD(20)) were performed repeatedly during the study. During FP-treatment, the severity of EIB decreased significantly as compared to placebo within 3 weeks, the geometric mean % fall in FEV(1) being reduced from 34.1% to 9.9% for 100 microg FP bid, and from 35.9% to 7.6% for 250 microg FP bid (P < 0.05). These reductions in EIB did not differ between the 2 doses and were sustained throughout the treatment period. PD(20) methacholine improved significantly during the first 6 weeks as compared to placebo (P < 0.04) and steadily increased with time in both treatment limbs (P = 0.04), the difference in improvement between doses (100 microg FP bid, 1.6 dose steps; 250 microg FP bid, 3.3 dose steps) approaching significance after 24 weeks (P = 0.06). We conclude that in childhood asthma, the protection afforded by inhaled fluticasone propionate against methacholine-induced bronchoconstriction is time- and dose-dependent, whereas protection against EIB is not. This suggests different modes of action of inhaled steroids in protecting against these pharmacological and physiological stimuli. This has to be taken into account when monitoring asthma treatment. PMID- 10821722 TI - One-year safety study with bambuterol once daily and terbutaline three times daily in 2-12-year-old children with asthma. The Bambuterol Multicentre Study Group. AB - The aim of this study was to compare bambuterol oral solution (10 mg) administered once daily in the evening with terbutaline oral solution (0.075 mg/kg body weight) administered three times daily in 2-5-year-old children and to compare bambuterol tablets (10 mg or 20 mg) administered once daily in the evening with terbutaline tablets (2.5 mg) administered three times daily in 6-12 year-old children with asthma. The study was of an open, randomized, parallel group design, and lasted 1 year. The primary objective was to evaluate safety (pulse rate, blood pressure, adverse events, hematology, and clinical chemistry). Plasma terbutaline concentrations were also measured. Evaluation of efficacy (FEV(1)) was a secondary objective. A total of 141 patients (83 boys, 58 girls) were randomized and treated with the study drugs, i.e., 43 patients in the terbutaline group (30 on oral solution and 13 on tablets) and 98 patients in the bambuterol group (62 on oral solution and 36 on tablets). A total of 11 patients discontinued the study: 3 were on terbutaline, and 8 were on bambuterol. There were no clinically important differences between treatment groups regarding pulse rate, or systolic or diastolic blood pressure. There were no clinically important findings in the laboratory tests (hematology and clinical chemistry). Both terbutaline and bambuterol were well-tolerated, and the reported adverse events were mostly mild or moderate. Mean steady state plasma terbutaline concentrations at the visits ranged between 8.0-11.5 nmol/L in the bambuterol tablet group and between 10.6-15.2 nmol/L in the terbutaline tablet group. The corresponding values in children on oral solution were 10.3-11.3 nmol/L in the bambuterol group and 7.5-9.7 nmol/L in the terbutaline group. FEV(1) measured in the 6-12-year-old children increased by more than 0.2 L in both treatment groups during the year in the study. In conclusion, bambuterol tablets or oral solution once daily and terbutaline tablets or oral solution three times daily showed a comparable and favorable safety profile. PMID- 10821723 TI - Exhaled nitric oxide is reduced after sputum induction in asthmatic children. AB - Exhaled nitric oxide (ENO) and eosinophil sputum markers are considered noninvasive markers of airway inflammation in asthma. The aim of this study was to evaluate whether the procedure of sputum induction can affect the level of ENO. We measured ENO before and after sputum induction by inhalation of hypertonic saline solution in 22 asthmatic children and 9 healthy controls. The ENO mean (+/- S. E.M.) value in the group of asthmatic children was reduced from a baseline value of 20.8 (+/- 3.0) ppb to 17.4 (+/- 2.4) ppb after sputum induction (P = 0.0012). In the healthy controls, the mean baseline value of ENO was 9.1 (+/- 2.1) ppb and it was reduced to 4. 8 (+/- 1.1) ppb after induction of sputum (P < 0.01). We suggest that measurements of ENO should be performed after the induction of sputum in asthmatic patients whenever both tests are done in sequence. PMID- 10821724 TI - Exhaled nitric oxide in children measured by tidal breathing method: differences between asthmatics and nonasthmatic controls. AB - The single-breath maneuver used to measure nitric oxide (NO) in adults cannot be performed by young children. We, therefore, developed a method of measuring NO in mixed exhaled gas collected during tidal breathing. NO was measured in mixed exhaled gas during 5 min of tidal breathing in 113 children 4-14 years of age: 22 nonasthmatics, 21 asthmatic children not taking inhaled corticosteroids, and 70 asthmatic children using inhaled corticosteroids. Exhaled NO levels (median, range) were significantly lower in nonasthmatic controls (median, range: 7, 2-10 ppb) than in asthmatic children on inhaled corticosteroids (8, 3-25 ppb; 95% CI for difference in medians with those of controls, 0-4 ppb), and in those not on inhaled corticosteroids (13, 6-37, ppb; 95% CI for difference in medians, 5-17 ppb). Asthmatic children not using inhaled corticosteroids had significantly higher exhaled NO levels than asthmatic children using inhaled corticosteroids (95% CI for difference in medians, 3-10 ppb). The tidal breathing method is a useful and practical way of measuring exhaled NO levels in children regardless of their age. PMID- 10821725 TI - Treatment of complicated pleural effusion with intracavitary urokinase in children. AB - Intrapleural administration of fibrinolytic agents such as urokinase (UK) has been advocated as an alternative method to manage complicated pleural effusion (CPE). Despite the increasing number of empyemas successfully treated with UK in adults, the experience in children is limited to a few cases. We report the results of image-guided catheter drainage (IGCD) with intracavitary instillation of UK in six children with CPE. Urokinase (25,000-100, 000 IU) was diluted in 20 mL of normal saline and instilled into the pleural cavity via a percutaneously placed drainage catheter. After 4 hr, the clamped catheter was released and connected to water-seal suction at a negative pressure of 20 cm H(2)O. UK instillation was repeated daily until no further drainage occurred. During IGCD, repeated radiographic and ultrasound imaging determined the location and amount of any remaining pleural fluid. Mean duration of hospital stay before initiating UK therapy was 4.3 days. Mean duration of catheter drainage before initiating UK therapy was 3.5 days, and the mean total drainage was 86 mL. All patients had an increase in chest tube drainage within 24 hr after the first instillation of UK. The mean net total drainage after UK instillation was 281 mL, most of the drainage being occurring in the first 2 days of treatment. Mean hospital stay following UK treatment was 5.8 days, and the average total duration of hospital stay was 13.8 days. No complications and no adverse events occurred during treatment with UK. Complete resolution of the consequences of the pleural effusion was observed in all patients at follow-up. Our results suggest that IGCD with adjunctive UK therapy is a reliable, simple, and safe approach to treat CPE, and it can reduce the risks associated with thoracotomy and decortication. PMID- 10821726 TI - Immunoglobulin levels in bronchoalveolar lavage fluid of children with chronic chest disease. AB - The concentration and distribution of immunoglobulin isotypes (IgG, IgM, sIgA) and IgG-subclass levels (IgG-1-4) were measured in bronchoalveolar lavage fluid (BALF) in 47 children with chronic chest disease (age range 1.0-9.9 years) and 18 healthy controls (age range 1.0-6.25 years). Of these patients, 19 had nonallergic asthma (Group A), 19 suffered from recurrent pneumonia or chronic bronchitis (Group B), and 9 patients had IgG-2 deficiency (Group C). In all individuals, IgG was the predominant immunoglobulin in the lower respiratory tract, followed by IgA and IgM. In patients of Group A and B, IgG, IgM and IgA levels in BALF were significantly elevated when compared to controls. Assessment of IgG-subclass concentrations in BALF revealed that IgG-1 levels were increased in Group A and B when compared to controls (P < 0.05). Since this difference could not be explained by difference in age, it is possibly due to the inflammatory process at the mucosal level. IgG-2 levels were elevated in all patients except those with IgG-2 deficiency. IgG-2 concentration in the IgG-2 deficent group was lower compared to controls (P < 0.005) and patients in Group A (P < 0.0005) and B (P < 0.005). IgG-3 levels were elevated in asthmatics in group A compared to healthy controls (P < 0.005). IgG-4 concentrations were the same in all study groups. Since IgG-subclasses in percentage of total IgG were similar in BALF and serum, our results do not indicate a local production of any of the IgG subclasses in the respiratory tract. PMID- 10821727 TI - Sweat testing following newborn screening for cystic fibrosis. AB - Sweat testing remains the "gold standard" for the diagnosis of cystic fibrosis (CF) and is a critical component of newborn screening programs. We retrospectively reviewed sweat test results reported to a neonatal screening program for CF with respect to completeness of reported results and the values recorded for sweat chloride (Cl(-)) and sodium (Na(+)) concentrations and the Cl( ):Na(+) ratio in screened infants. Thirty-nine of 85 DeltaF508 homozygous (DeltaF508/DeltaF508) and 270 of 274 DeltaF508 heterozygous (DeltaF508/-) infants had sweat tests reported to the screening program. Of those, 30 and 213 sweat test reports, respectively, were complete, i.e., sweat weight, sweat chloride, and sodium were reported. Three centers accounted for 37 of 68 (54%) incomplete results, and 4 centers performed 4 or less post-screening sweat tests in the study period. There were 6 DeltaF508 heterozygous infants with sweat Cl(-) concentrations of 40-60 mmol/L and 4 had CF confirmed by additional genotyping (n = 2) or clinical and repeat sweat Cl results (n = 2). Forty-one percent of DeltaF508/-infants with sweat Cl(-) <40 mmol/L had Cl:Na >1. We conclude that the reporting of incomplete sweat tests is common following newborn screening for CF. Infants with sweat Cl(-) levels of 40-60 mmol/L require further investigation and review, but they almost certainly have CF. The Cl(-):Na(+) ratio does not appear useful in establishing a diagnosis of CF in infants. PMID- 10821728 TI - Wisconsin cystic fibrosis chest radiograph scoring system: validation and standardization for application to longitudinal studies. AB - This study was designed to achieve a final modeling, validation, and standardization plan for the Wisconsin cystic fibrosis (CF) chest radiographic scoring system. Sixty chest radiographs were selected to reflect a range of severity of lung pathology in children with CF. Seven experienced volunteer raters (three radiologists and four pediatric pulmonologists) from five institutions were recruited to evaluate and score the films. Analysis of scores revealed that the subcomponents of the Wisconsin system showed considerable variation from rater to rater, but reliability assessment indicated satisfactory Cronbach's alpha coefficients (0.83-0.90) among the seven raters. It was found that an additive method of total score computation is significantly more reliable (P < 0.05) than either the original multiplicative model or the traditional Brasfield scoring system. Comparison of radiologists and pulmonologists revealed a marked, systematic difference in scoring with the former group being more conservative in interpretation of abnormalities than the pulmonologists, and some of the raters showing very limited sensitivity. Quantitative chest radiology applied to children with cystic fibrosis studied long-term in longitudinal research projects requires the careful use of sensitive scoring methods and careful selection and training of multiple raters. This is particularly important since pulmonologists and radiologists can differ systematically in interpreting/scoring abnormalities. PMID- 10821729 TI - Sniff nasal inspiratory pressure in children. AB - Maximal static inspiratory pressure (P(IMAX)) generated at the mouth is one of the tests of inspiratory muscle strength in children. In adults, inspiratory muscle strength has also been assessed using sniff nasal inspiratory pressure (SNIP). This test is easier to perform than P(IMAX) and might therefore be applicable to children. To test this hypothesis, we measured P(IMAX) and SNIP in 116 children aged 4 to 11 years (54 girls, 62 boys). P(IMAX) was measured using a tube mouthpiece and SNIP using a tightly fitting plug in one nostril, while a sniff was performed through the other nostril. Both tests were performed from functional residual capacity (FRC). Pressure was measured with a differential pressure transducer and displayed in real time on a computer screen. Weight, standing height, sitting height, gender, and age were recorded. There was a significant difference (P < 0.01) in group mean (SD) data between SNIP (81.3 (27.4) cmH(2)O) and P(IMAX) (67.9 (28.1) cmH(2)O). Bland/Altman analysis demonstrated a mean difference of -13.5 cmH(2)O (SD 21.4) between the techniques. Regression analysis indicated highly significant relations (P < 0.01) between SNIP and P(IMAX), and between weight, standing and sitting height, and age for SNIP, and between weight, standing height, and age for P(IMAX). SNIP and P(IMAX) were greater in boys than girls (83.2 vs. 79.2 cmH(2)O SNIP; 72.9 vs. 62.0 cmH(2)O P(IMAX)), but this difference was only significant for P(IMAX) (P < 0.05). SNIP was significantly greater than P(IMAX) (P < 0.01) in both boys and girls. These data suggest that SNIP provides a simple, noninvasive additional test to P(IMAX) for assessing inspiratory muscle strength in children. PMID- 10821730 TI - Recurrent lower respiratory tract infections in a 14-year-old boy with tracheobronchomegaly (Mounier-Kuhn syndrome). AB - Tracheobronchomegaly (Mounier-Kuhn syndrome) is characterized by dilatation of the central airways, tracheobronchial outpouchings, and chronic tracheobronchitis. Most cases are diagnosed in adulthood. We report the clinical, radiographic, and bronchoscopic findings in a 14-year-old boy with tracheobronchomegaly, ptosis of the right eyelid, and redundant skin and mucosa of the upper lip, who presented with a 2-year history of recurrent lower respiratory tract infections. Pediatricians should be aware of the possibility that tracheobronchomegaly may cause clinical symptoms in childhood and adolescence. Pediatr Pulmonol. 2000; 29: 476-479. PMID- 10821731 TI - Twenty-four-hour esophageal pH-monitoring and wheezing. PMID- 10821732 TI - Gastroesophageal reflux in infants with wheezing. PMID- 10821734 TI - Selected abstracts PMID- 10821733 TI - The authors respond PMID- 10821735 TI - Cops and robbers: putative evolution of copper oxygen-binding proteins. AB - Two closely related copper proteins, phenoloxidase and haemocyanin, are known to be involved in different physiological functions such as the primary immune response and oxygen transport. Although the proteins differ structurally, they have the same active site by which dioxygen is bound. Recent results reveal that haemocyanin also exhibits phenoloxidase activity. A scenario is proposed for the evolutionary relationships among copper oxygen-binding proteins (COPs). PMID- 10821736 TI - The macrophage in acute neural injury: changes in cell numbers over time and levels of cytokine production in mammalian central and peripheral nervous systems. AB - We evaluated the timing and density of ED-1-positive macrophage accumulation (ED 1 is the primary antibody for the macrophage) and measured cytokine production by macrophages in standardized compression injuries to the spinal cord and sciatic nerves of individual rats 3, 5, 10 and 21 days post-injury. The actual site of mechanical damage to the nervous tissue, and a more distant site where Wallerian degeneration had occurred, were evaluated in both the peripheral nervous system (PNS) and the central nervous system (CNS) at these time points. The initial accumulation of activated macrophages was similar at both the central and peripheral sites of damage. Subsequently, macrophage densities at all locations studied were statistically significantly higher in the spinal cord than in the sciatic nerve at every time point but one. The peak concentrations of three cytokines, tumor necrosis factor &agr; (TNF &agr; ), interleukin-1 (IL-1) and interleukin-6 (IL-6), appeared earlier and were statistically significantly higher in injured spinal cord than in injured sciatic nerve. We discuss the meaning of these data relative to the known differences in the reparative responses of the PNS and CNS to injury. PMID- 10821737 TI - Central circuitry in the jellyfish Aglantha digitale. III. The rootlet and pacemaker systems. AB - Tactile stimulation of the subumbrella of Aglantha digitale was found to evoke an escape swimming response similar to that evoked by stimulation of the outer surfaces of the margin but that does not involve the ring giant axon. Evidence is presented that conduction around the margin takes place via an interconnected system of rootlet interneurones. Confocal microscopy of carboxyfluorescein-filled axons showed that the rootlet neurones run out from the bases of the motor giant axons within the inner nerve ring and come into close contact with those of the neighbouring motor giant axons on either side. Transmission between the rootlet neurones has the properties of chemical synaptic transmission. A distinct type of fast excitatory postsynaptic potential (rootlet PSP) was recorded in motor giant axons following stimulation of nearby axons in 3-5 mmol l(-)(1) Mn(2+), which lowered the PSP below spike threshold. Immune labelling with anti-syntaxin 1 showed structures tentatively identified as synapses in the inner nerve ring, including some on the rootlet neurones. Neuromuscular junctions were not labelled. A secondary consequence of stimulating motor giant axons was the triggering of events in the pacemaker system. Triggering was blocked in 105 mmol l(-)(1) Mg(2+), indicating a synaptic link. Activity in the pacemaker system led indirectly to tentacle contractions (as described in earlier papers in this series), but the contractions were not as sudden or as violent as those seen when escape swimming was mediated by the ring giant axon. Events triggered in the pacemaker system fed back into the motor giants, producing postsynaptic potentials that appeared as humps in the spike after-potential. The conduction velocity of events propagating in the relay system was increased when the rootlet pathway was simultaneously excited (piggyback effect). With the addition of the rootlet pathway, the number of identified systems concerned with locomotion, feeding and tentacle contractions comes to fourteen, and the list is probably nearly complete. PMID- 10821738 TI - Activity before exercise influences recovery metabolism in the lizard Dipsosaurus dorsalis. AB - During recovery from even a brief period of exercise, metabolic rate remains elevated above resting levels for extended periods. The intensity and duration of exercise as well as body temperature and hormone levels can influence this excess post-exercise oxygen consumption (EPOC). We examined the influence of activity before exercise (ABE), commonly termed warm-up in endotherms, on EPOC in the desert iguana Dipsosaurus dorsalis. The rate of oxygen consumption and blood lactate levels were measured in 11 female D. dorsalis (mass 41.1 +/- 3.0 g; mean +/- s.e.m.) during rest, after two types of ABE and after 5 min of exhaustive exercise followed by 60 min of recovery. ABE was either single (15 s of maximal activity followed by a 27 min pause) or intermittent (twelve 15 s periods of exercise separated by 2 min pauses). Our results indicate that both single and intermittent ABE reduced recovery metabolic rate. EPOC volumes decreased from 0.261 to 0.156 ml of oxygen consumed during 60 min of recovery when lizards were subjected to intermittent ABE. The average cost of activity (net V(O2) during exercise and 60 min of recovery per distance traveled) was almost 40 % greater in lizards that exercised without any prior activity than in lizards that underwent ABE. Blood lactate levels and removal rates were greatest in animals that underwent ABE. These findings may be of particular importance for terrestrial ectotherms that typically use burst locomotion and have a small aerobic scope and a long recovery period. PMID- 10821739 TI - Tissue-specific expression of zebrafish (Danio rerio) heat shock factor 1 mRNAs in response to heat stress. AB - All organisms respond to environmental, chemical and physiological stresses by enhanced synthesis of an evolutionarily conserved family of proteins known as heat shock proteins (HSPs) or stress proteins. Certain HSPs are also expressed constitutively during cell growth and development, and they function as molecular chaperones. The transcriptional regulation of hsp genes is mediated by the heat shock transcription factor (HSF). The stress response has been studied mostly in mammalian cell lines or organisms normally maintained under constant laboratory conditions. There is much less information on the regulation of the stress response of animals, such as fish, that have to tolerate large fluctuations in environmental and internal conditions. To characterize the regulation of the heat shock response in fish, we have cloned the first heat shock transcription factor from fish, zebrafish Danio rerio. Phylogenetic analysis confirms that the isolated zebrafish HSF belongs to the HSF1 family and is therefore designated zHSF1. Analysis by reverse transcriptase polymerase chain reaction (RT-PCR) shows the presence of two zHSF1 mRNA forms that are expressed in a tissue-specific fashion upon exposure to heat stress. Both forms are expressed in gonads under all conditions; in liver and to a lesser extent in the gills, the longer splice form of zHSF1 disappears upon heat shock. We present evidence for a unique tissue specific regulation of HSF1 upon exposure to elevated temperature. PMID- 10821740 TI - Behavioural thermoregulation of chum salmon during homing migration in coastal waters. AB - Homing Pacific salmon are suggested to utilise directional cues in shallow water while migrating in coastal waters. Since salmonids are typical cold-water fish, they may have to cope with warm surface water while gathering directional information. We studied behavioural thermoregulation of 31 free-ranging chum salmon Oncorhynchus keta using micro data loggers off the Sanriku coast from early October to December. The surface water temperature was approximately 20 degrees C in early October and decreased to approximately 12 degrees C in December. The seasonal change of water temperature transformed the behaviour of salmon markedly from deep diving to shallow swimming. In October, salmon frequently dived to depths exceeding 100 m. Duration of deep dives tended to be prolonged as the thermal difference between sea surface and bottom water increased. The results indicated that salmon sought the coolest thermal refuge that they could exploit by vertical movement. Thermal refuge could be a way for salmon to minimise metabolic energy cost; however, salmon repeatedly returned to the surface water column. We found a positive correlation between surface swimming of salmon and the presence of cool covering water, which could contain river waters. This suggests that salmon tend to be risk-prone when subjected to a high concentration of directional cues. Salmon in December spent most of their time in shallow water. These findings suggest that salmon adjusted their behavioural strategy with the hydrographic structure of the sea in order to achieve a balance between acquiring directional cues and behavioural thermoregulation. PMID- 10821741 TI - Ion channel activity from the midgut brush-border membrane of gypsy moth (Lymantria dispar) larvae. AB - Ion channels from the midgut apical membrane of gypsy moth (Lymantria dispar) larvae were studied following mechanical fusion of brush-border membrane vesicles with planar phospholipid bilayer membranes. In symmetrical 300 mmol l(-)(1) KCl (pH 9.0), nine different channels with conductances ranging from 27 to 795 pS and linear current/voltage relationships were resolved. In the presence of a KCl gradient across the bilayer (450 mmol l(-)(1 )cis/150 mmol l(-)(1 )trans), 11 different conductance levels ranging from 16 to 850 pS were detected. The channels were slightly cationic: the zero-current reversal potential was shifted by -5 mV to -21 mV compared with symmetrical KCl conditions, corresponding to p(K)/p(Cl) permeability ratios of 1.5-8.0. Most channels were neither voltage dependent nor Ca(2+)-sensitive and displayed complex gating kinetics. Addition of Ba(2+) or Cs(+) to both sides of the bilayer had little effect on channel activity, but fewer distinct channels were observed when KCl was replaced by potassium gluconate, suggesting an effect of Cl(-) on channel activity. A reduced number of channels was also detected when KCl was replaced by N-methyl-d glucamine-HCl. Under asymmetrical N-methyl-d-glucamine-HCl conditions, only anionic channels were observed. They exhibited current rectification (35 pS at negative voltages and 81 pS at positive voltages) and were strongly voltage dependent. PMID- 10821742 TI - Vocal tract function in birdsong production: experimental manipulation of beak movements. AB - Kinematic analyses have demonstrated that the extent to which a songbird's beak is open when singing correlates with the acoustic frequencies of the sounds produced, suggesting that beak movements function to modulate the acoustic properties of the vocal tract during song production. If motions of the beak are necessary for normal song production, then disrupting the ability of a bird to perform these movements should alter the acoustic properties of its song. We tested this prediction by comparing songs produced normally by white-throated sparrows and swamp sparrows with songs produced when the beak was temporarily immobilized. We also observed how temporarily loading the beak of canaries with extra mass affected vocal tract movements and song production. Disruption of vocal tract movements resulted in the predicted frequency-dependent amplitude changes in the songs of both white-throated sparrows and swamp sparrows. Canaries with mass added to their beak sang with their beak open more widely than normal and produced notes with greater harmonic content than those without weights. Both manipulations resulted in acoustic changes consistent with a model in which beak motions affect vocal tract resonances, thus supporting the hypothesis that dynamic vocal tract motions and post-production modulation of sound are necessary features of normal song production. PMID- 10821743 TI - Responses of young Xenopus laevis tadpoles to light dimming: possible roles for the pineal eye. AB - When the light is dimmed, the pineal eye of hatchling Xenopus laevis tadpoles excites the central pattern generator for swimming, but the behavioural significance of pineal excitation is unclear. We show that tadpoles spend 99 % of their time hanging from the surface meniscus or solid objects using mucus secreted by a cement gland on the head. Attachment inhibits swimming, but unattached tadpoles swim spontaneously. Provided that their pineal eye is intact, they attach closer to the water surface in the dark than in the light and attach preferentially to the underside of floating objects that cast shadows. Dimming causes tadpoles swimming horizontally to turn upwards and is very effective in initiating upward swimming in unattached tadpoles. Similar pineal-dependent responses during swimming are present up to stage 44. Pinealectomy blocks responses to dimming at all stages. Recordings from immobilised tadpoles reveal that light dimming induces faster fictive swimming and that pineal activity is increased for up to 20 min during sustained light dimming. We suggest that the increase in pineal discharge during dimming increases the probability of upward swimming and, in this way, increases the probability of tadpoles attaching to objects higher in the water column that cast shadows. PMID- 10821744 TI - Simple mechanisms organise orientation of escape swimming in embryos and hatchling tadpoles of Xenopus laevis. AB - Many amphibian tadpoles hatch and swim before their inner ears and sense of spatial orientation differentiate. We describe upward and downward swimming responses in hatchling Xenopus laevis tadpoles from stages 32 to 37/38 in which the body rotates about its longitudinal axis. Tadpoles are heavier than water and, if touched while lying on the substratum, they reliably swim upwards, often in a tight spiral. This response has been observed using stroboscopic photography and high-speed video recordings. The sense of the spiral is not fixed for individual tadpoles. In 'more horizontal swimming' (i.e. in directions within +/ 30 degrees of the horizontal), the tadpoles usually swim belly-down, but this position is not a prerequisite for subsequent upward spiral swimming. Newly hatched tadpoles spend 99 % of their time hanging tail-down from mucus secreted by a cement gland on the head. When suspended in mid-water by a mucus strand, tadpoles from stage 31 to 37/38 tend to swim spirally down when touched on the head and up when touched on the tail. The three-dimensional swimming paths of stage 33/34 tadpoles were plotted using simultaneous video images recorded from the side and from above. Tadpoles spiralled for 70 % of the swimming time, and the probability of spiralling increased to 1 as swim path angles became more vertical. Tadpoles were neutrally buoyant in Percoll/water mixtures at 1.05 g cm( )(3), in which anaesthetised tadpoles floated belly-down and head-up at 30 degrees. In water, their centre of mass was ventral to the muscles in the yolk mass. A simple mathematical model suggests that the orientation of tadpoles during swimming is governed by the action of two torques, one of which raises the head (i.e. increases the pitch) and the other rotates (rolls) the body. Consequently, tadpoles (i) swim belly-down when the body is approximately horizontal because the body is ballasted by dense yolk, and (ii) swim spirally at more vertical orientations when the ballasting no longer stabilises orientation. Measurements in tethered tadpoles show that dorsal body flexion, which could produce a dorsal pitch torque, is present during swimming and increases with tailbeat frequency. We discuss how much of the tadpole's behaviour can be explained by our mathematical model and suggest that, at this stage of development, oriented swimming responses may depend on simple touch reflexes, the organisation of the muscles and physical features of the body, rather than on vestibular reflexes. PMID- 10821745 TI - Adhesion measured on the attachment pads of Tettigonia viridissima (Orthoptera, insecta). AB - The tarsi of the cricket Tettigonia viridissima bear flexible attachment pads that are able to deform, replicating the profile of a surface to which they are apposed. This attachment system is supplemented by a secretion produced by epidermal cells and transported onto the surface of the pad through the pore canals of the pad cuticle. This study shows that the secretion alone is necessary, but not sufficient, for adhesion. To account for the full adhesive force, the deformation of the pad and the resulting changes in contact area were considered. In two series of experiments, the adhesive properties of the secretion and the adhesion of the whole pad were measured using a force tester, the sensitivity of which ranged from micronewtons to centinewtons. The adhesive forces of the secretion measured between a smooth sapphire ball with a diameter of 1.47 mm and a flat silicon surface ranged from 0.1 to 0.6 mN. In a control experiment on the silicon surface without secretion, no adhesive force was measured. There was no dependence of the adhesive force on the applied compressive force. When an intact pad was pulled off a flat silicon surface, the adhesive force increased with increasing applied compressive force, but it did not increase further once the applied force exceeded a certain value. The saturated adhesive force, ranging from 0.7 to 1.2 mN, was obtained at applied forces of 0.7-1.5 mN. The hemispherical surface of the pad had a larger contact area and demonstrated greater adhesion under a larger applied force. Adhesion became saturated when a pad was deformed such that contact area was maximal. The tenacity (the adhesive force per unit area) was 1.7-2.2 mN mm(-)(2). PMID- 10821746 TI - Control of epithelial Cl(-) secretion by basolateral osmolality in the euryhaline teleost Fundulus heteroclitus. AB - Euryhaline teleost fish adapt rapidly to salinity change and reduce their rate of ion secretion on entry to fresh water. Killifish (Fundulus heteroclitus) transferred from full-strength sea water to fresh water showed large reductions in plasma [Na(+)] and osmolality at 6 h which were corrected by 24 h. To mimic this in vitro, a hypotonic shock of 20-70 mosmol kg(-)(1) was applied on the basolateral side of opercular epithelia. This hypotonic shock reversibly reduced the short-circuit current (I(sc), equivalent to the rate of secretion of Cl(-)) in a dose-dependent fashion, with a 40 mosmol kg(-)(1) hypotonic shock reducing I(sc) by 58+/-4.6 % in 40 min. Similar reductions in [NaCl], but with added mannitol to maintain osmolality, were without effect, indicating that the effect was purely osmotic. Hypotonic inhibition of I(sc) was accompanied by reductions in epithelial conductance (G(t)) but no significant change in transepithelial potential (V(t)). The hypotonic inhibition was apparently not Ca(2+)-mediated because Ca(2+)-depleted salines, thapsigargin and ionomycin all failed to block the reduction in I(sc) produced by hypotonic shock. The inhibition was not mediated via a reduction in intracellular cyclic AMP level because cyclic AMP levels, measured by radioimmunoassay, were unchanged by hypotonic shock and by 1.0 micromol l(-)(1) clonidine (which inhibits I(sc) by changing intracellular [Ca(2+)]) but were increased markedly by 1.0 micromol l(-)(1) isoproterenol, a positive control. The protein tyrosine kinase inhibitor genistein (100 micromol l(-)(1)), but not its inactive analogue daidzein, inhibited I(sc) in normal osmolality but produced a stimulation of I(sc) after hypotonic shock (and after clonidine treatment). The inhibitory effects of genistein and hypotonicity were not additive, suggesting that the same portion of the I(sc) was inhibited by both treatments. These data are consistent with a model for Cl(-) transport regulation involving tyrosine phosphorylation in cell-swelling-induced inhibition of Cl(-) secretion when euryhaline teleosts adapt to fresh water. PMID- 10821747 TI - Skin temperatures during free-ranging swimming and diving in antarctic fur seals. AB - This study tests the hypothesis that an endothermic homeotherm should minimise heat flux in cold polar waters by minimising skin temperature. Temperature variability was measured at the surface of the skin of three Antarctic fur seals (Arctocephalus gazella) at intervals of 2 s over a total of 9.7 days while they were swimming and diving freely in polar waters at temperatures of 1.5-4 degrees C. The temperature difference ( capdelta T) between skin on the dorsal thorax and the water varied from more than 20 degrees C to close to equality over periods of less than 1 h. Shorter-term variations in capdelta T of up to 5 degrees C occurred in association with diving, although these types of variations also occurred without diving. In general, capdelta T began to decline during the descent phase of a dive and began to increase again during the ascent or at the end of the dive. One of the three individuals examined showed little variation in capdelta T, which remained low (approximately 3 degrees C) throughout the experiment. In the other two fur seals, capdelta T tended to decline during periods of sustained diving and usually increased during periods spent at the surface. Mean calculated heat flux varied from 95 to 236 W m(-)(2) depending on the individual. Metabolic rates based on these calculated heat fluxes were towards the lower end of those measured in previous studies using different methodologies. The study has shown that Antarctic fur seal skin temperature is highly dynamic and suggests that the thoracic surface is an organ used for active thermoregulation. PMID- 10821748 TI - Hydrodynamic drag in steller sea lions (Eumetopias jubatus). AB - Drag forces acting on Steller sea lions (Eumetopias jubatus) were investigated from 'deceleration during glide' measurements. A total of 66 glides from six juvenile sea lions yielded a mean drag coefficient (referenced to total wetted surface area) of 0.0056 at a mean Reynolds number of 5.5x10(6). The drag values indicate that the boundary layer is largely turbulent for Steller sea lions swimming at these Reynolds numbers, which are past the point of expected transition from laminar to turbulent flow. The position of maximum thickness (at 34 % of the body length measured from the tip of the nose) was more anterior than for a 'laminar' profile, supporting the idea that there is little laminar flow. The Steller sea lions in our study were characterized by a mean fineness ratio of 5.55. Their streamlined shape helps to delay flow separation, reducing total drag. In addition, turbulent boundary layers are more stable than laminar ones. Thus, separation should occur further back on the animal. Steller sea lions are the largest of the otariids and swam faster than the smaller California sea lions (Zalophus californianus). The mean glide velocity of the individual Steller sea lions ranged from 2.9 to 3.4 m s(-)(1) or 1.2-1.5 body lengths s(-)(1). These length-specific speeds are close to the optimum swim velocity of 1.4 body lengths s(-)(1) based on the minimum cost of transport for California sea lions. PMID- 10821749 TI - Vertebrate ancient (VA) opsin and extraretinal photoreception in the Atlantic salmon (Salmo salar). AB - A member of a new photopigment family first isolated from teleost fish, vertebrate ancient (VA) opsin, has recently been shown to form a functional photopigment and to be expressed within a subset of horizontal and amacrine cells of the inner retina. These sites of expression (and structural features) of VA opsin suggest that this photopigment might mediate non-image-forming light detection tasks. We attempted to gain support for this hypothesis by examining the expression of VA opsin within the central nervous system (CNS) (pineal and deep brain) of the Atlantic salmon Salmo salar. In addition, we examined the sites of rod-opsin, cone-opsin and &agr; -transducin expression within the salmon CNS to provide a more complete description of the extraretinal photoreceptors of a teleost vertebrate. We show that multiple populations of cells within the salmon CNS appear to contain photoreceptors: VA opsin was strongly expressed in the pineal organ and in bilateral columns of subependymal cells in the epithalamus; anti-cone-opsin antibodies labelled cells within the pineal and numerous cells in the anterior hypothalamus (suprachiasmatic nucleus, nucleus preopticus magnocellularis, nucleus preopticus parvocellularis); anti-rod-opsin antibodies labelled cells within the pineal but no other areas within the central brain; and anti- &agr; -transducin antibodies labelled cells within the pineal and the ventral telencephalon. Collectively, our results suggest that VA opsin is a photopigment specialised for irradiance detection tasks within the eye, pineal and central brain, and that the salmon has multiple and varied populations of photoreceptors within the CNS. We review the significance of these findings within the broad context of vertebrate extraretinal photoreception. PMID- 10821750 TI - Allometric scaling relationships of jumping performance in the striped marsh frog Limnodynastes peronii. AB - We constructed a force platform to investigate the scaling relationships of the detailed dynamics of jumping performance in striped marsh frogs (Limnodynastes peronii). Data were used to test between two alternative models that describe the scaling of anuran jumping performance; Hill's model, which predicts mass- independence of jump distance, and Marsh's model, which predicts that jump distance increases as M(0.2), where M is body mass. From the force platform, scaling relationships were calculated for maximum jumping force (F(max)), acceleration, take-off velocity (U(max)), mass-specific jumping power (P(max)), total jumping distance (D(J)) and total contact time for 75 L. peronii weighing between 2.9 and 38. 4 g. F(max) was positively correlated with body mass and was described by the equation F(max)=0.16M(0.61), while P(max) decreased significantly with body mass and was described by the equation P(max)=347M( )(0.46). Both D(J) and U(max) were mass-independent over the post-metamorph size range, and thus more closely resembled Hill's model for the scaling of locomotion. We also examined the scaling relationships of jumping performance in metamorph L. peronii by recording the maximum jump distance of 39 animals weighing between 0.19 and 0.58 g. In contrast to the post-metamorphic L. peronii, D(J) and U(max) were highly dependent on body mass in metamorphs and were described by the equations D(J)=38M(0.53) and U(max)=1.82M(0.23), respectively. Neither model for the scaling of anuran jumping performance resembled data from metamorph L. peronii. Although the hindlimbs of post-metamorphic L. peronii scaled geometrically (body mass exponent approximately 0.33), the hindlimbs of metamorphs showed greater proportional increases with body mass (mass exponents of 0.41-0.42). PMID- 10821751 TI - Glial cell fate specification modulated by the bHLH gene Hes5 in mouse retina. AB - Neurons and glial cells differentiate from common precursors. Whereas the gene glial cells missing (gcm) determines the glial fate in Drosophila, current data about the expression patterns suggest that, in mammals, gcm homologues are unlikely to regulate gliogenesis. Here, we found that, in mouse retina, the bHLH gene Hes5 was specifically expressed by differentiating Muller glial cells and that misexpression of Hes5 with recombinant retrovirus significantly increased the population of glial cells at the expense of neurons. Conversely, Hes5 deficient retina showed 30-40% decrease of Muller glial cell number without affecting cell survival. These results indicate that Hes5 modulates glial cell fate specification in mouse retina. PMID- 10821752 TI - Fate restriction in limb muscle precursor cells precedes high-level expression of MyoD family member genes. AB - The mechanisms by which pluripotent embryonic cells generate unipotent tissue progenitor cells during development are unknown. Molecular/genetic experiments in cultured cells have led to the hypothesis that the product of a single member of the MyoD gene family (MDF) is necessary and sufficient to establish the positive aspects of the determined state of myogenic precursor cells: i.e., the ability to initiate and maintain the differentiated state (Weintraub, H., Davis, R., Tapscott, S., Thayer, M., Krause, M., Benezra, R., Blackwell, T. K., Turner, D., Rupp, R., Hollenberg, S. et al. (1991) Science 251, 761-766). Embryonic cell type determination also involves negative regulation, such as the restriction of developmental potential for alternative cell types, that is not directly addressed by the MDF model. In the experiments reported here, phenotypic restriction in myogenic precursor cells is assayed by an in vivo 'notochord challenge' to evaluate their potential to 'choose' between two alternative cell fate endpoints: cartilage and muscle (Williams, B. A. and Ordahl, C. P. (1997) Development 124, 4983-4997). Two separate myogenic precursor cell populations were found to be phenotypically restricted while expressing the Pax3 gene and prior to MDF gene activation. Therefore, while MDF family members act positively during myogenic differentiation, phenotypic restriction, the negative aspect of cell specification, requires cellular and molecular events and interactions that precede MDF expression in myogenic precursor cells. The qualities of muscle formed by the determined myogenic precursor cells in these experiments further indicate that their developmental potential is intermediate between that of myoblastic stem cells taken from fetal or adult tissue (which lack mitotic and morphogenetic potential when tested in vivo) and embryonic stem cells (which are multipotent). We hypothesize that such embryonic myogenic progenitor cells represent a distinct class of determined embryonic cell, one that is responsible for both tissue growth and tissue morphogenesis. PMID- 10821753 TI - Querkopf, a MYST family histone acetyltransferase, is required for normal cerebral cortex development. AB - In order to find, and mutate, novel genes required for regulation of neurogenesis in the cerebral cortex, we performed a genetic screen in mice. As the result of this screen, we created a new mouse mutant, querkopf. The querkopf mutation is due to an insertion into a MYST family histone acetyltransferase gene. Mice homozygous for the querkopf mutation have craniofacial abnormalities, fail to thrive in the postnatal period and have defects in central nervous system development. The defects in central nervous system development are particularly prominent in the cerebral cortex, which is disproportionally smaller than in wild type mice. A large reduction in the size of the cortical plate was already apparent during embryogenesis. Homozygous mice show a lack of large pyramidal cells in layer V of the cortex, which is reflected in a reduction in the number of Otx1-positive neurons in this layer during postnatal development. Homozygous mice also show a reduction in the number of GAD67-positive interneurons throughout the cortex. Our results suggest that Querkopf is an essential component of a genetic cascade regulating cell differentiation in the cortex, probably acting in a multiprotein complex regulating chromatin structure during transcription. PMID- 10821754 TI - Ace/Fgf8 is required for forebrain commissure formation and patterning of the telencephalon. AB - Fibroblast growth factors (Fgfs) form a large family of secreted signalling proteins that have a wide variety of roles during embryonic development. Within the central nervous system (CNS) Fgf8 is implicated in patterning neural tissue adjacent to the midbrain-hindbrain boundary. However, the roles of Fgfs in CNS tissue rostral to the midbrain are less clear. Here we examine the patterning of the forebrain in zebrafish embryos that lack functional Fgf8/Ace. We find that Ace is required for the development of midline structures in the forebrain. In the absence of Ace activity, midline cells fail to adopt their normal morphology and exhibit altered patterns of gene expression. This disruption to midline tissue leads to severe commissural axon pathway defects, including misprojections from the eye to ectopic ipsilateral and contralateral targets. Ace is also required for the differentiation of the basal telencephalon and several populations of putative telencephalic neurons but not for overall regional patterning of forebrain derivatives. Finally, we show that ace expression co localises with anterior neural plate cells that have previously been shown to have forebrain patterning activity. Removal of these cells leads to a failure in induction of ace expression indicating that loss of Ace activity may contribute to the phenotypes observed when anterior neural plate cells are ablated. However, as ace mutant neural plate cells still retain at least some inductive activity, then other signals must also be produced by the anterior margin of the neural plate. PMID- 10821755 TI - FGF10 is an inducer and Pax6 a competence factor for lacrimal gland development. AB - We investigated the mechanism of tissue induction and specification using the lacrimal gland as a model system. This structure begins its morphogenesis as a bud-like outgrowth of the conjunctival epithelium and ultimately forms a branched structure with secretory function. Using a reporter transgene as a specific marker for gland epithelium, we show that the transcription factor Pax6 is required for normal development of the gland and is probably an important competence factor. In investigating the cell-cell signaling required, we show that fibroblast growth factor (FGF) 10 is sufficient to stimulate ectopic lacrimal bud formation in ocular explants. Expression of FGF10 in the mesenchyme adjacent to the presumptive lacrimal bud and absence of lacrimal gland development in FGF10-null mice strongly suggest that it is an endogenous inducer. This was supported by the observation that inhibition of signaling by a receptor for FGF10 (receptor 2 IIIb) suppressed development of the endogenous lacrimal bud. In explants of mesenchyme-free gland epithelium, FGF10 stimulated growth but not branching morphogenesis. This suggested that its role in induction is to stimulate proliferation and, in turn, that FGF10 combines with other factors to provide the instructive signals required for lacrimal gland development. PMID- 10821758 TI - Presenilin-1 regulates neuronal differentiation during neurogenesis. AB - Mutations in Presenilin-1 (PS1) are a major cause of familial Alzheimer's disease. Our previous studies showed that PS1 is required for murine neural development. Here we report that lack of PS1 leads to premature differentiation of neural progenitor cells, indicating a role for PS1 in a cell fate decision between postmitotic neurons and neural progenitor cells. Neural proliferation and apoptotic cell death during neurogenesis are unaltered in PS1(-/-) mice, suggesting that the reduction in the neural progenitor cells observed in the PS1( /-) brain is due to premature differentiation of progenitor cells, rather than to increased apoptotic cell death or decreased cell proliferation. In addition, the premature neuronal differentiation in the PS1(-/-) brain is associated with aberrant neuronal migration and disorganization of the laminar architecture of the developing cerebral hemisphere. In the ventricular zone of PS1(-/-) mice, expression of the Notch1 downstream effector gene Hes5 is reduced and expression of the Notch1 ligand Dll1 is elevated, whereas expression of Notch1 is unchanged. The level of Dll1 transcripts is also increased in the presomitic mesoderm of PS1(-/-) embryos, while the level of Notch1 transcripts is unchanged, in contrast to a previous report (Wong et al., 1997, Nature 387, 288-292). These results provide direct evidence that PS1 controls neuronal differentiation in association with the downregulation of Notch signalling during neurogenesis. PMID- 10821756 TI - The bHLH transcription factor hand2 plays parallel roles in zebrafish heart and pectoral fin development. AB - The precursors of several organs reside within the lateral plate mesoderm of vertebrate embryos. Here, we demonstrate that the zebrafish hands off locus is essential for the development of two structures derived from the lateral plate mesoderm - the heart and the pectoral fin. hands off mutant embryos have defects in myocardial development from an early stage: they produce a reduced number of myocardial precursors, and the myocardial tissue that does form is improperly patterned and fails to maintain tbx5 expression. A similar array of defects is observed in the differentiation of the pectoral fin mesenchyme: small fin buds form in a delayed fashion, anteroposterior patterning of the fin mesenchyme is absent and tbx5 expression is poorly maintained. Defects in these mesodermal structures are preceded by the aberrant morphogenesis of both the cardiogenic and forelimb-forming regions of the lateral plate mesoderm. Molecular analysis of two hands off alleles indicates that the hands off locus encodes the bHLH transcription factor Hand2, which is expressed in the lateral plate mesoderm starting at the completion of gastrulation. Thus, these studies reveal early functions for Hand2 in several cellular processes and highlight a genetic parallel between heart and forelimb development. PMID- 10821757 TI - bozozok and squint act in parallel to specify dorsal mesoderm and anterior neuroectoderm in zebrafish. AB - In vertebrate embryos, maternal (beta)-catenin protein activates the expression of zygotic genes that establish the dorsal axial structures. Among the zygotically acting genes with key roles in the specification of dorsal axial structures are the homeobox gene bozozok (boz) and the nodal-related (TGF-(beta) family) gene squint (sqt). Both genes are expressed in the dorsal yolk syncytial layer, a source of dorsal mesoderm inducing signals, and mutational analysis has indicated that boz and sqt are required for dorsal mesoderm development. Here we examine the regulatory interactions among boz, sqt and a second nodal-related gene, cyclops (cyc). Three lines of evidence indicate that boz and sqt act in parallel to specify dorsal mesoderm and anterior neuroectoderm. First, boz requires sqt function to induce high levels of ectopic dorsal mesoderm, consistent with sqt acting either downstream or in parallel to boz. Second, sqt mRNA is expressed in blastula stage boz mutants, indicating that boz is not essential for activation of sqt transcription, and conversely, boz mRNA is expressed in blastula stage sqt mutants. Third, boz;sqt double mutants have a much more severe phenotype than boz and sqt single mutants. Double mutants consistently lack the anterior neural tube and axial mesoderm, and ventral fates are markedly expanded. Expression of chordin and noggin1 is greatly reduced in boz;sqt mutants, indicating that the boz and sqt pathways have overlapping roles in activating secreted BMP antagonists. In striking contrast to boz;sqt double mutants, anterior neural fates are specified in boz;sqt;cyc triple mutants. This indicates that cyc represses anterior neural development, and that boz and sqt counteract this repressive function. Our results support a model in which boz and sqt act in parallel to induce dorsalizing BMP-antagonists and to counteract the repressive function of cyc in neural patterning. PMID- 10821759 TI - Integrins modulate the Egfr signaling pathway to regulate tendon cell differentiation in the Drosophila embryo. AB - Changes in the extracellular matrix (ECM) govern the differentiation of many cell types during embryogenesis. Integrins are cell matrix receptors that play a major role in cell-ECM adhesion and in transmitting signals from the ECM inside the cell to regulate gene expression. In this paper, it is shown that the PS integrins are required at the muscle attachment sites of the Drosophila embryo to regulate tendon cell differentiation. The analysis of the requirements of the individual alpha subunits, alphaPS1 and alphaPS2, demonstrates that both PS1 and PS2 integrins are involved in this process. In the absence of PS integrin function, the expression of tendon cell-specific genes such as stripe and beta1 tubulin is not maintained. In addition, embryos lacking the PS integrins also exhibit reduced levels of activated MAPK. This reduction is probably due to a downregulation of the Epidermal Growth Factor receptor (Egfr) pathway, since an activated form of the Egfr can rescue the phenotype of embryos mutant for the PS integrins. Furthermore, the levels of the Egfr ligand Vein at the muscle attachment sites are reduced in PS mutant embryos. Altogether, these results lead to a model in which integrin-mediated adhesion plays a role in regulating tendon cell differentiation by modulating the activity of the Egfr pathway at the level of its ligand Vein. PMID- 10821760 TI - Impaired migration and delayed differentiation of pancreatic islet cells in mice lacking EGF-receptors. AB - Pancreatic acini and islets are believed to differentiate from common ductal precursors through a process requiring various growth factors. Epidermal growth factor receptor (EGF-R) is expressed throughout the developing pancreas. We have analyzed here the pancreatic phenotype of EGF-R deficient (-/-) mice, which generally die from epithelial immaturity within the first postnatal week. The pancreata appeared macroscopically normal. The most striking feature of the EGF-R (-/-) islets was that instead of forming circular clusters, the islet cells were mainly located in streak-like structures directly associated with pancreatic ducts. Based on BrdU-labelling, proliferation of the neonatal EGF-R (-/-) beta cells was significantly reduced (2.6+/-0.4 versus 5.8+/-0.9%, P<0.01) and the difference persisted even at 7-11 days of age. Analysis of embryonic pancreata revealed impaired branching morphogenesis and delayed islet cell differentiation in the EGF-R (-/-) mice. Islet development was analyzed further in organ cultures of E12.5 pancreata. The proportion of insulin-positive cells was significantly lower in the EGF-R (-/-) explants (27+/-6 versus 48+/-8%, P<0.01), indicating delayed differentiation of the beta cells. Branching of the epithelium into ducts was also impaired. Matrix metalloproteinase (MMP-2 and MMP-9) activity was reduced 20% in EGF-R (-/-) late-gestation pancreata, as measured by gelatinase assays. Furthermore, the levels of secreted plasminogen activator inhibitor-1 (PAI-1) were markedly higher, while no apparent differences were seen in the levels of active uPA and tPa between EGF-R (-/-) and wild-type pancreata. Our findings suggest that the perturbation of EGF-R-mediated signalling can lead to a generalized proliferation defect of the pancreatic epithelia associated with a delay in beta cell development and disturbed migration of the developing islet cells as they differentiate from their precursors. Upregulated PAI-1 production and decreased gelatinolytic activity correlated to this migration defect. An intact EGF-R pathway appears to be a prerequisite for normal pancreatic development. PMID- 10821761 TI - Rac1 mutations produce aberrant epithelial differentiation in the developing and adult mouse small intestine. AB - The mouse small intestinal epithelium undergoes continuous renewal throughout life. Previous studies suggest that differentiation of this epithelium is regulated by instructions that are received as cells migrate along crypt-villus units. The nature of the instructions and their intracellular processing remain largely undefined. In this report, we have used genetic mosaic analysis to examine the role of Rac1 GTPase-mediated signaling in controlling differentiation. A constitutively active mutation (Rac1Leu61) or a dominant negative mutation (Rac1Asn17) was expressed in the 129/Sv embryonic stem cell derived component of the small intestine of C57Bl/6-ROSA26<->129/Sv mice. Rac1Leu61 induces precocious differentiation of members of the Paneth cell and enterocytic lineages in the proliferative compartment of the fetal gut, without suppressing cell division. Forced expression of the dominant negative mutation inhibits epithelial differentiation, without affecting cell division, and slows enterocytic migration along crypt-villus units. The effects produced by Rac1Leu61 or Rac1Asn17 in the 129/Sv epithelium do not spread to adjacent normal C57Bl/6 epithelial cells. These results provide in vivo evidence that Rac1 is involved in the import and intracellular processing of signals that control differentiation of a mammalian epithelium. PMID- 10821762 TI - Blastocyst H(2) receptor is the target for uterine histamine in implantation in the mouse. AB - The process of implantation is a 'two-way' interaction between the blastocyst and uterus. It has long been suspected that histamine is an important mediator in embryo-uterine interactions during implantation, but its source, targets and mechanism of actions remained undefined. We have recently demonstrated that uterine epithelial cells are the source of histamine, which peaks on day 4 of pregnancy (the day of implantation) in the mouse. In searching for its target and site of action, we discovered that preimplantation blastocysts, which express histamine type 2 receptor (H(2)), is the target for histamine action. Using multiple approaches, we demonstrate herein that uterine-derived histamine interacts with embryonic H(2) receptors in a paracrine fashion to initiate the process of implantation. PMID- 10821763 TI - Mutations in the stumpy gene reveal intermediate targets for zebrafish motor axons. AB - Primary motoneurons, the earliest developing spinal motoneurons in zebrafish, have highly stereotyped axon projections. Although much is known about the development of these neurons, the molecular cues guiding their axons have not been identified. In a screen designed to reveal mutations affecting motor axons, we isolated two mutations in the stumpy gene that dramatically affect pathfinding by the primary motoneuron, CaP. In stumpy mutants, CaP axons extend along the common pathway, a region shared by other primary motor axons, but stall at an intermediate target, the horizontal myoseptum, and fail to extend along their axon-specific pathway during the first day of development. Later, most CaP axons progress a short distance beyond the horizontal myoseptum, but tend to stall at another intermediate target. Mosaic analysis revealed that stumpy function is needed both autonomously in CaP and non-autonomously in other cells. stumpy function is also required for axons of other primary and secondary motoneurons to progress properly past intermediate targets and to branch. These results reveal a series of intermediate targets involved in motor axon guidance and suggest that stumpy function is required for motor axons to progress from proximally located intermediate targets to distally located ones. PMID- 10821764 TI - The retinal determination gene, dachshund, is required for mushroom body cell differentiation. AB - The dachshund gene of Drosophila encodes a putative transcriptional regulator required for eye and leg development. We show here that dachshund is also required for normal brain development. The mushroom bodies of dachshund mutants exhibit a marked reduction in the number of (&agr;) lobe axons, a disorganization of axons extending into horizontal lobes, and aberrant projections into brain areas normally unoccupied by mushroom body processes. The phenotypes become pronounced during pupariation, suggesting that dachshund function is required during this period. GAL4-mediated expression of dachshund in the mushroom bodies rescues the mushroom body phenotypes. Moreover, dachshund mutant mushroom body clones in an otherwise wild-type brain exhibit the phenotypes, indicating an autonomous role for dachshund. Although eyeless, like dachshund, is preferentially expressed in the mushroom body and is genetically upstream of dachshund for eye development, no interaction of these genes was detected for mushroom body development. Thus, dachshund functions in the developing mushroom body neurons to ensure their proper differentiation. PMID- 10821765 TI - Axon routing at the optic chiasm after enzymatic removal of chondroitin sulfate in mouse embryos. AB - The effects of removing chondroitin sulfate from chondroitin sulfate proteoglycan molecules on guidance of retinal ganglion cell axons at the optic chiasm were investigated in a brain slice preparation of mouse embryos of embryonic day 13 to 15. Slices were grown for 5 hours and growth of dye-labeled axons was traced through the chiasm. After continuous enzymatic digestion of the chondroitin sulfate proteoglycans with chondroitinase ABC, which removes the glycosaminoglycan chains, navigation of retinal axons was disrupted. At embryonic day 13, before the uncrossed projection forms in normal development, many axons deviated from their normal course, crossing the midline at aberrant positions and invading the ventral diencephalon. In slices from embryonic day 14 embryos, axons that would normally form the uncrossed projection at this stage failed to turn into the ipsilateral optic tract. In embryonic day 15 slices, enzyme treatment caused a reduction of the uncrossed projection that develops at this stage. Growth cones in enzyme-treated slices showed a significant increase in the size both before and after they crossed the midline. This indicates that responses of retinal axons to guidance signals at the chiasm have changed after removal of the chondroitin sulfate epitope. We concluded that the chondroitin sulfate moieties of the proteoglycans are involved in patterning the early phase of axonal growth across the midline and at a later stage controlling the axon divergence at the chiasm. PMID- 10821766 TI - Function and regulation of homothorax in the wing imaginal disc of Drosophila. AB - The gene homothorax (hth) is originally expressed uniformly in the wing imaginal disc but, during development, its activity is restricted to the cells that form the thorax and the hinge, where the wing blade attaches to the thorax, and eliminated in the wing pouch, which forms the wing blade. We show that hth repression in the wing pouch is a prerequisite for wing development; forcing hth expression prevents growth of the wing blade. Both the Dpp and the Wg pathways are involved in hth repression. Cells unable to process the Dpp (lacking thick veins or Mothers against Dpp activity) or the Wg (lacking dishevelled function) signal express hth in the wing pouch. We have identified vestigial (vg) as a Wg and Dpp response factor that is involved in hth control. In contrast to its repressing role in the wing pouch, wg upregulates hth expression in the hinge. We have also identified the gene teashirt (tsh) as a positive regulator of hth in the hinge. tsh plays a role specifying hinge structures, possibly in co-operation with hth. PMID- 10821767 TI - Bmp4 and Fgf10 play opposing roles during lung bud morphogenesis. AB - Morphogenesis of the mouse lung involves reciprocal interactions between the epithelial endoderm and the surrounding mesenchyme, leading to an invariant early pattern of branching that forms the basis of the respiratory tree. There is evidence that Fibroblast growth factor 10 (Fgf10) and Bone Morphogenetic Protein 4 (Bmp4), expressed in the distal mesenchyme and endoderm, respectively, play important roles in branching morphogenesis. To examine these roles in more detail, we have exploited an in vitro culture system in which isolated endoderm is incubated in Matrigel(TM) substratum with Fgf-loaded beads. In addition, we have used a Bmp4(lacZ) line of mice in which lacZ faithfully reports Bmp4 expression. Analysis of lung endoderm in vivo shows a dynamic pattern of Bmp4(lacZ) expression during bud outgrowth, extension and branching. In vitro, Fgf10 induces both proliferation and chemotaxis of isolated endoderm, whether it is derived from the distal or proximal lung. Moreover, after 48 hours, Bmp4(lacZ) expression is upregulated in the endoderm closest to the bead. Addition of 30-50 ng/ml of exogenous purified Bmp4 to the culture medium inhibits Fgf-induced budding or chemotaxis, and inhibits overall proliferation. By contrast, the Bmp binding protein Noggin enhances Fgf-induced morphogenesis. Based on these and other results, we propose a model for the combinatorial roles of Fgf10 and Bmp4 in branching morphogenesis of the lung. PMID- 10821768 TI - Analysis of the promoter of the cudA gene reveals novel mechanisms of Dictyostelium cell type differentiation. AB - The cudA gene encodes a nuclear protein that is essential for normal multicellular development. At the slug stage cudA is expressed in the prespore cells and in a sub-region of the prestalk zone. We show that cap site distal promoter sequences direct cudA expression in prespore cells, while proximal sequences direct expression in the prestalk sub-region. The promoter domain that directs prespore-specific transcription consists of a positively acting region, that has the potential to direct expression in all cells within the slug, and a negatively acting region that prevents expression in the prestalk cells. Dd-STATa is the STAT protein that regulates commitment to stalk cell gene expression, where it is known to function as a transcriptional repressor. We show that Dd STATa binds in vitro to the positively acting part of the prespore domain of the cudA promoter. However, Dd-STATa cannot be utilised for this purpose in vivo, because analysis of a Dd-STATa null mutant strain shows that Dd-STATa is not necessary for cudA transcription in prespore cells. In contrast, the part of the cudA promoter that directs prestalk-specific expression contains a binding site for Dd-STATa that is essential for its biological activity. Dd-STATa appears therefore to serve as a direct activator of cudA transcription in prestalk cells, while a protein with a DNA binding specificity highly related to that of Dd-STATa is utilised to activate cudA transcription in prespore cells. PMID- 10821769 TI - Three-dimensional in vivo analysis of Dictyostelium mounds reveals directional sorting of prestalk cells and defines a role for the myosin II regulatory light chain in prestalk cell sorting and tip protrusion. AB - During cell sorting in Dictyostelium, we observed that GFP-tagged prestalk cells (ecmAO-expressing cells) moved independently and directionally to form a cluster. This is consistent with a chemotaxis model for cell sorting (and not differential adhesion) in which a long-range signal attracts many of the prestalk cells to the site of cluster formation. Surprisingly, the ecmAO prestalk cluster that we observed was initially found at a random location within the mound of this Ax3 strain, defining an intermediate sorting stage not widely reported in Dictyostelium. The cluster then moved en masse to the top of the mound to produce the classic, apical pattern of ecmAO prestalk cells. Migration of the cluster was also directional, suggesting the presence of another long-range guidance cue. Once at the mound apex, the cluster continued moving upward leading to protrusion of the mound's tip. To investigate the role of the cluster in tip protrusion, we examined ecmAO prestalk-cell sorting in a myosin II regulatory light chain (RLC) null in which tips fail to form. In RLC-null mounds, ecmAO prestalk cells formed an initial cluster that began to move to the mound apex, but then arrested as a vertical column that extended from the mound's apex to its base. Mixing experiments with wild-type cells demonstrated that the RLC-null ecmAO prestalk cell defect is cell autonomous. These observations define a specific mechanism for myosin's function in tip formation, namely a mechanical role in the upward movement of the ecmAO prestalk cluster. The wild-type data demonstrate that cell sorting can occur in two steps, suggesting that, in this Ax3 strain, spatially and temporally distinct cues may guide prestalk cells first to an initial cluster and then later to the tip. PMID- 10821770 TI - Region-specific activation of the Xenopus brachyury promoter involves active repression in ectoderm and endoderm: a study using transgenic frog embryos. AB - Tissue specification in the early embryo requires the integration of spatial information at the promoters of developmentally important genes. Although several response elements for signalling pathways have been identified in Xenopus promoters, it is not yet understood what defines the sharp borders that restrict expression to a specific tissue. Here we use transgenic frog embryos to study the spatial and temporal regulation of the Xbra promoter. Deletion analysis and point mutations in putative transcription factor-binding sites identified two repressor modules, which exert their main effects at different stages during gastrulation. One module is defined by a bipartite binding site for a Smad-interacting protein (SIP1) of the deltaEF1 repressor family and acts to confine expression to the marginal zone early in gastrulation. The other module is defined by two homeodomain-binding sites and is responsible for repression in dorsal mesoderm and ectoderm at mid-gastrula stages. In addition, an upstream region of the promoter is necessary to repress expression in neural tissues later in development. Together, our results show that repression plays an important role in the restriction of Xbra expression to the mesoderm, and we suggest that similar mechanisms may be involved in the spatial regulation of other genes in early embryonic development. PMID- 10821771 TI - Isolation of chicken vasa homolog gene and tracing the origin of primordial germ cells. AB - To obtain a reliable molecular probe to trace the origin of germ cell lineages in birds, we isolated a chicken homolog (Cvh) to vasa gene (vas), which plays an essential role in germline formation in Drosophila. We demonstrate the germline specific expression of CVH protein throughout all stages of development. Immunohistochemical analyses using specific antibody raised against CVH protein indicated that CVH protein was localized in cytoplasm of germ cells ranging from presumptive primordial germ cells (PGCs) in uterine-stage embryos to spermatids and oocytes in adult gonads. During the early cleavages, CVH protein was restrictively localized in the basal portion of the cleavage furrow. About 30 CVH expressing cells were scattered in the central zone of the area pellucida at stage X, later 45-60 cells were found in the hypoblast layer and subsequently 200 250 positive cells were found anteriorly in the germinal crescent due to morphogenetic movement. Furthermore, in the oocytes, CVH protein was predominantly localized in granulofibrillar structures surrounding the mitochondrial cloud and spectrin protein-enriched structure, indicating that the CVH-containing cytoplasmic structure is the precursory germ plasm in the chicken. These results strongly suggest that the chicken germline is determined by maternally inherited factors in the germ plasm. PMID- 10821772 TI - Analysis of cranial neural crest migratory pathways in axolotl using cell markers and transplantation. AB - We have examined the ability of normal and heterotopically transplanted neural crest cells to migrate along cranial neural crest pathways in the axolotl using focal DiI injections and in situ hybridization with the neural crest marker, AP 2. DiI labeling demonstrates that cranial neural crest cells migrate as distinct streams along prescribed pathways to populate the maxillary and mandibular processes of the first branchial arch, the hyoid arch and gill arches 1-4, following migratory pathways similar to those observed in other vertebrates. Another neural crest marker, the transcription factor AP-2, is expressed by premigratory neural crest cells within the neural folds and migrating neural crest cells en route to and within the branchial arches. Rotations of the cranial neural folds suggest that premigratory neural crest cells are not committed to a specific branchial arch fate, but can compensate when displaced short distances from their targets by migrating to a new target arch. In contrast, when cells are displaced far from their original location, they appear unable to respond appropriately to their new milieu such that they fail to migrate or appear to migrate randomly. When trunk neural folds are grafted heterotopically into the head, trunk neural crest cells migrate in a highly disorganized fashion and fail to follow normal cranial neural crest pathways. Importantly, we find incorporation of some trunk cells into branchial arch cartilage despite the random nature of their migration. This is the first demonstration that trunk neural crest cells can form cartilage when transplanted to the head. Our results indicate that, although cranial and trunk neural crest cells have inherent differences in ability to recognize migratory pathways, trunk neural crest can differentiate into cranial cartilage when given proper instructive cues. PMID- 10821773 TI - Hedgehog signals regulate multiple aspects of gastrointestinal development. AB - The gastrointestinal tract develops from the embryonic gut, which is composed of an endodermally derived epithelium surrounded by cells of mesodermal origin. Cell signaling between these two tissue layers appears to play a critical role in coordinating patterning and organogenesis of the gut and its derivatives. We have assessed the function of Sonic hedgehog and Indian hedgehog genes, which encode members of the Hedgehog family of cell signals. Both are expressed in gut endoderm, whereas target genes are expressed in discrete layers in the mesenchyme. It was unclear whether functional redundancy between the two genes would preclude a genetic analysis of the roles of Hedgehog signaling in the mouse gut. We show here that the mouse gut has both common and separate requirements for Sonic hedgehog and Indian hedgehog. Both Sonic hedgehog and Indian hedgehog mutant mice show reduced smooth muscle, gut malrotation and annular pancreas. Sonic hedgehog mutants display intestinal transformation of the stomach, duodenal stenosis (obstruction), abnormal innervation of the gut and imperforate anus. Indian hedgehog mutants show reduced epithelial stem cell proliferation and differentiation, together with features typical of Hirschsprung's disease (aganglionic colon). These results show that Hedgehog signals are essential for organogenesis of the mammalian gastrointestinal tract and suggest that mutations in members of this signaling pathway may be involved in human gastrointestinal malformations. PMID- 10821774 TI - Gadolinium inhibits Na(+)-Ca(2+) exchanger current in guinea-pig isolated ventricular myocytes. AB - The trivalent cation, gadolinium (Gd(3+)) is commonly used to inhibit stretch activated channels. In this report, we show that Gd(3+) also inhibits ionic current (I(NaCa)), carried by the Na(+)-Ca(2+) exchanger protein. Under selective recording conditions, Gd(3+) inhibited both outward and inward I(NaCa) from guinea-pig isolated ventricular myocytes in a dose-dependent manner, with half maximal inhibition concentrations (IC(50)) of 30.0+/-4.0 microM at +60 mV (Hill coefficient, h=1.04+/-0.13) and 20.0+/-2.7 microM at -100 mV (h=1.13+/-0.16), respectively (P>0.05, n=5 - 9). Thus, inhibition was not voltage-dependent. The time from Gd(3+) application to steady-state effect was slow compared to the divalent blocker Ni(2+). The slow time course appeared to reflect gradual Gd(3+) accumulation at its binding site on the exchanger, rather than a use-dependent blocking mechanism. This study indicates that for experiments in which Gd(3+) is used, its inhibitory effect on I(NaCa) should be taken into account. PMID- 10821775 TI - Pharmacological characterization of neurogenic responses of the sheep isolated internal anal sphincter. AB - The aim of the study was to establish the nature of the neurogenic responses of the sheep isolated anal sphincter. Isolated strips of sheep internal anal sphincter develop intrinsic contractile tone following the application of stretch tension. On transmural stimulation (1 - 20 Hz, 10 V pulse strength, 0.5 ms pulse width, 1 s every 180 s) transient relaxations were observed. The amplitude of the relaxations were frequency-dependent reaching a maximal response at 10 - 20 Hz and were inhibited by tetrodotoxin (0.3 microM). Neither atropine (0.3 microM) nor phentolamine (1 microM) affected control responses. The nitric oxide synthase inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME, 100 microM) and the selective inhibitor of soluble guanylyl cyclase ODQ, (1H-[1,2, 4]oxadiazolo[4,3 a]quinoxalin-1-one) (1 microM) completely inhibited the neurogenic relaxations and uncovered contractions that were abolished by 1 microM phentolamine and 0.1 microM prazosin. The effect of L-NAME, but not that of ODQ, was partially reversed by the addition of L-arginine (1 mM). Sodium nitroprusside (10 nM - 10 microM) caused concentration-dependent inhibition of myogenic tone and this effect was significantly reduced by ODQ. Calcium-free Krebs-Henseleit solution also reduced myogenic tone by 85%. Transmural electrical stimulation of the sheep isolated internal anal sphincter causes a transient relaxation of myogenic tone that appears to involve nitric oxide from non-adrenergic, non-cholinergic nerves and, to a lesser degree, noradrenaline from sympathetic nerves. The characteristics of the preparation compares well with that of human tissue and may prove to be a suitable animal based model for further studies. PMID- 10821776 TI - Effects of peroxisome proliferator-activated receptor-alpha and -gamma agonist, JTT-501, on diabetic complications in Zucker diabetic fatty rats. AB - This study has investigated the effects of JTT-501, a peroxisome proliferator activated receptor (PPAR)-alpha and PPAR-gamma agonist, on the pathogenesis of diabetic complications in the Zucker diabetic fatty (ZDF) rats, a model of type 2 diabetes. Comparison is made with troglitazone, a PPAR-gamma agonist. The ZDF rats exhibited hyperglycaemia and hyperlipidaemia, and developed diabetic complications such as cataract, nephropathy, and neuropathy. Treatment with JTT 501 from the prediabetic stage controlled glycaemia and lipidaemia, and prevented the development of diabetic complications. Troglitazone was less effective in controlling serum cholesterol and neuropathy. ZDF rats developed diabetic osteopenia with reduced bone turnover, and this was prevented by JTT-501 and troglitazone, possibly mediated by increased bone turnover and bone formation. Since JTT-501 controlled glycaemia and lipidaemia in ZDF rats and prevented several diabetic complications, it is suggested that treatment with JTT-501, which activates both PPAR-alpha and PPAR-gamma, could provide a valuable therapeutic approach against diabetic complications in type 2 diabetes. PMID- 10821777 TI - Substance P mediates inflammatory oedema in acute pancreatitis via activation of the neurokinin-1 receptor in rats and mice. AB - Pancreatic oedema occurs early in the development of acute pancreatitis, and the overall extent of fluid loss correlates with disease severity. The tachykinin substance P (SP) is released from sensory nerves, binds to the neurokinin-1 receptor (NK1-R) on endothelial cells and induces plasma extravasation, oedema, and neutrophil infiltration, a process termed neurogenic inflammation. We sought to determine the importance of neurogenic mechanisms in acute pancreatitis. Pancreatic plasma extravasation was measured using the intravascular tracers Evans blue and Monastral blue after administration of specific NK1-R agonists/antagonists in rats and NK1-R(+/+)/(-/-) mice. The effects of NK1-R genetic deletion/antagonism on pancreatic plasma extravasation, amylase, myeloperoxidase (MPO), and histology in cerulein-induced pancreatitis were characterized. In rats, both SP and the NK1-R selective agonist [Sar(9) Met(O(2))(11)]SP stimulated pancreatic plasma extravasation, and this response was blocked by the NK1-R antagonist CP 96,345. Selective agonists of the NK-2 or NK-3 receptors had no effect. In rats, cerulein stimulated pancreatic plasma extravasation and serum amylase. These responses were blocked by the NK1-R antagonist CP 96,345. In wildtype mice, SP induced plasma extravasation while SP had no effect in NK1-R knockout mice. In NK1-R knockout mice, the effects of cerulein on pancreatic plasma extravasation and hyperamylasemia were reduced by 60%, and pancreatic MPO by 75%, as compared to wildtype animals. Neurogenic mechanisms of inflammation are important in the development of inflammatory oedema in acute interstitial pancreatitis. PMID- 10821778 TI - Antagonist effects on human P2X(7) receptor-mediated cellular accumulation of YO PRO-1. AB - We have examined the interaction of P2 antagonists with the human P2X(7) receptor by studying their effect on 2' and 3'-O-benzoyl-benzoyl-ATP (DbATP) stimulated cellular accumulation of the fluorescent, DNA binding dye, YO-PRO-1 (MW=375Da). In suspensions of HEK293 cells expressing human recombinant P2X(7) receptors, DbATP produced time and concentration-dependent increases in YO-PRO-1 fluorescence. This response presumably reflects YO-PRO-1 entry through P2X(7) receptor channels and binding to nucleic acids. When studies were performed in a NaCl-free, sucrose-containing buffer, full concentration-effect curves to DbATP could be constructed. The P2 antagonists, pyridoxalphosphate-6-azophenyl-2', 4' disulphonic acid (PPADS) and periodate oxidized ATP (oATP), reduced the potency of DbATP and decreased its maximum response. 1-[N,O-bis(1, 5 isoquinolinesulphonyl)-N-methyl-L-tyrosyl]-4-phenylpiperazine (KN62) and its analogue, KN04, reduced the potency of DbATP. Schild slopes for KN62 and KN04 were shallow and exhibited a plateau at concentrations of compound greater than 1 microM, indicating that these compounds were not competitive antagonists. Calmidazolium and a monoclonal antibody to human P2X(7) receptors attenuated DbATP-stimulated YO-PRO-1 accumulation but they were not competitive antagonists and only produced 2 - 3 fold decreases in the potency of DbATP. The effects of PPADS and KN62 were partially reversible whereas those of oATP were not. PPADS protected cells against the irreversible antagonist effects of oATP suggesting a common site of action. In contrast KN62 was not effective suggesting that it may bind at a different site to oATP and PPADS. This study has demonstrated that P2X(7) receptor function can be quantified by measuring DbATP stimulated YO-PRO-1 accumulation and has provided additional information about the interaction of P2 receptor antagonists with the human P2X(7) receptor. PMID- 10821779 TI - Tissue angiotensin II and endothelin-1 modulate differently the response to flow in mesenteric resistance arteries of normotensive and spontaneously hypertensive rats. AB - In resistance arteries pressure-induced (myogenic) tone (MT) and flow (shear stress)-induced dilation (FD) are potent determinant of vascular resistance. We investigated the role of angiotensin II and endothelin-1 in FD and MT in resistance arteries and their potential change in hypertension. Flow - diameter - pressure relationship was established in situ, under anaesthesia, in two daughter branches of a mesenteric resistance artery (180 microM, n=7 per group) from spontaneously hypertensive (SHR) or normotensive (WKY) rats. One artery was ligated distally, so that it was submitted to pressure only, while the other was submitted to pressure and flow. Drugs were added to the preparation and external diameter, pressure and flow measured continuously. External diameter (with flow) ranged from 150+/-3 to 191+/-7 microM in WKY (n=28) rats and from 168+/-6 to 186+/-6 microM in SHR (n=28). Flow induced a dilation of the non-ligated arteries which was lower in SHR (13+/-5 - 31+/-4 microM vs WKY: 5+/-5 - 44+/-4 microM). In the ligated artery, the diameter did not significantly change, due to MT. In the vessels submitted to flow angiotensin converting enzyme inhibition (perindopril, 10 micromol L(-1)) increased the diameter in SHR (+11+/-2 microM) significantly more than in WKY (+2+/-1 microM). Angiotensin type 1 receptor (AT(1)R) blockade (losartan, 10 micromol L(-1)) increased the diameter in the vessels with flow in SHR only (+6+/-1 microM). Angiotensin type 2 receptor (AT(2)R) blockade (PD 123319, 1 micromol L(-1)) decreased arterial diameter in WKY only (9+/-2). Endothelin-1 type A receptor (ET(A)R) blockade (LU135252, 0.1 micromol L(-1)) increased the diameter only in SHR in the artery submitted to flow (by 6+/-1 microM). Thus FD was counteracted by a flow-dependent AT(1) and ET(A) receptors activation in SHR whereas in WKY FD AT(2)-dependent dilation is involved. PMID- 10821780 TI - Exploration of the ligand binding site of the human 5-HT(4) receptor by site directed mutagenesis and molecular modeling. AB - Among the five human 5-HT(4) (h5-HT(4)) receptor isoforms, the h5-HT(4(a)) receptor was studied with a particular emphasis on the molecular interactions involved in ligand binding. For this purpose, we used site-directed mutagenesis of the transmembrane domain. Twelve mutants were constructed with a special focus on the residue P4.53 of helix IV which substitutes in h5-HT(4) receptors the highly conserved S residue among the rhodopsin family receptors. The mutated receptors were transiently expressed in COS-7 cells. Ligand binding or competition studies with two h5-HT(4) receptor agonists, serotonin and ML10302 and two h5-HT(4) receptor antagonists, [(3)H]-GR113808 and ML10375 were performed on wild type and mutant receptors. Functional activity of the receptors was evaluated by measuring the ability of serotonin to stimulate adenylyl cyclase. Ligand binding experiments revealed that [(3)H]-GR113808 did not bind to mutants P4.53A, S5.43A, F6.51A, Y7.43A and to double mutant F6.52V/N6.55L. On the other hand mutations D3.32N, S5.43A and Y7.43A appeared to promote a dramatic decrease of h5-HT(4(a)) receptor functional activity. From these studies, S5.43 and Y7.43 clearly emerged as common anchoring sites to antagonist [(3)H]-GR113808 and to serotonin. According to these results, we propose ligand-receptor complex models with serotonin and [(3)H]-GR113808. For serotonin, three interaction points were selected including ionic interaction with D3.32, a stabilizing interaction of this ion pair by Y7.43 and a hydrogen bond with S5.43. [(3)H]-GR113808 was also docked, based on the same type of interactions with S5.43 and D3.32: the proposed model suggested a possible role of P4.53 in helix IV structure allowing the involvement of a close hydrophobic residue, W4.50, in a hydrophobic pocket for hydrophobic interactions with the indole ring of [(3)H]-GR113808. PMID- 10821781 TI - Characterization of SB-269970-A, a selective 5-HT(7) receptor antagonist. AB - The novel 5-HT(7) receptor antagonist, SB-269970-A, potently displaced [(3)H]-5 CT from human 5-HT(7(a)) (pK(i) 8.9+/-0.1) and 5-HT(7) receptors in guinea-pig cortex (pK(i) 8.3+/-0.2). 5-CT stimulated adenylyl cyclase activity in 5 HT(7(a))/HEK293 membranes (pEC(50) 7.5+/-0.1) and SB-269970-A (0.03 - 1 microM) inhibited the 5-CT concentration-response with no significant alteration in the maximal response. The pA(2) (8.5+/-0.2) for SB-269970-A agreed well with the pK(i) determined from [(3)H]-5-CT binding studies. 5-CT-stimulated adenylyl cyclase activity in guinea-pig hippocampal membranes (pEC(50) of 8.4+/-0.2) was inhibited by SB-269970-A (0.3 microM) with a pK(B) (8.3+/-0.1) in good agreement with its antagonist potency at the human cloned 5-HT(7(a)) receptor and its binding affinity at guinea-pig cortical membranes. 5-HT(7) receptor mRNA was highly expressed in human hypothalamus, amygdala, thalamus, hippocampus and testis. SB-269970-A was CNS penetrant (steady-state brain : blood ratio of ca. 0.83 : 1 in rats) but was rapidly cleared from the blood (CLb=ca. 140 ml min(-1) kg(-1)). Following a single dose (3 mg kg(-1)) SB-269970 was detectable in rat brain at 30 (87 nM) and 60 min (58 nM). In guinea-pigs, brain levels averaged 31 and 51 nM respectively at 30 and 60 min after dosing, although the compound was undetectable in one of the three animals tested. 5-CT (0.3 mg kg(-1) i.p.) induced hypothermia in guinea-pigs was blocked by SB-269970-A (ED(50) 2.96 mg kg( 1) i.p.) and the non-selective 5-HT(7) receptor antagonist metergoline (0.3 - 3 mg kg(-1) s.c.), suggesting a role for 5-HT(7) receptor stimulation in 5-CT induced hypothermia in guinea-pigs. SB-269970-A (30 mg kg(-1)) administered at the start of the sleep period, significantly reduced time spent in Paradoxical Sleep (PS) during the first 3 h of EEG recording in conscious rats. PMID- 10821782 TI - Mechanisms underlying the attenuation of endothelium-dependent vasodilatation in the mesenteric arterial bed of the streptozotocin-induced diabetic rat. AB - Experiments were designed to investigate the mechanisms underlying the diabetes related impairment of the vasodilatations of the perfused mesenteric arterial bed induced by acetylcholine (ACh) and K(+). In streptozotocin (STZ)-diabetic rats, the ACh-induced endothelium-dependent vasodilatation was attenuated. The dose response curves for ACh in control and diabetic rats were each shifted to the right by N(G)-nitro-L-arginine (L-NOARG) and by isotonic high K(+) (60 mM). The ACh dose-response curves under isotonic high K(+) were not different between control and diabetic rats. We also examined the vasodilatation induced by K(+), which is a putative endothelium-derived hyperpolarizing factor (EDHF). The mesenteric vasodilatation induced by a single administration of K(+) was greatly impaired in STZ-induced diabetic rats. Treatment with charybdotoxin plus apamin abolished the ACh-induced vasodilatation but enhanced the K(+)-induced response in controls and diabetic rats. After pretreatment with ouabain plus BaCl(2), the ACh-induced vasodilatation was significantly impaired and the K(+)-induced relaxation was abolished in both control and diabetic rats. The impairment of the endothelium-dependent vasodilatation of the mesenteric arterial bed seen in STZ induced diabetic rats may be largely due to a defective vascular response to EDHF. It is further suggested that K(+) is one of the endothelium-derived hyperpolarizing factors and that the vasodilatation response to K(+) is impaired in the mesenteric arterial bed from diabetic rats. PMID- 10821783 TI - Somatostatin potentiates NMDA receptor function via activation of InsP(3) receptors and PKC leading to removal of the Mg(2+) block without depolarization. AB - N-methyl-D-aspartate (NMDA) receptors exist on noradrenergic axon terminals and mediate enhancement of noradrenaline (NA) release. We here investigated modulation by somatostatin (SRIF, somatotropin release inhibiting factor) of the NMDA-induced release of NA using superfused hippocampal synaptosomes. The NMDA response was increased by SRIF-28 and SRIF-14, but not SRIF-28((1 - 14)), whereas the release of [(3)H]-NA elicited by alpha-amino-3-hydroxy-5-methylisoxazide-4 propionic acid (AMPA) was unaffected. SRIF-14 did not mimic glycine at the NMDA receptor but activated SRIF receptors sited on noradrenergic terminals. The SRIF 14 effect was blocked by pertussis toxin but mimicked by mastoparan, a G-protein activator. BIM-23056, but not Cyanamid 154806, antagonized the SRIF-14 effect. This effect was mimicked by L362855, a partial agonist at the sst(5) subtype, but not by the new selective sst(1) - sst(4) receptor agonists L797591, L779976, L796778 and L803087. Protein kinase C (PKC) inhibitors (H7, staurosporine, GF 209103X, cheleritrine and sphingosine) prevented the SRIF-14 effect, while phorbol 12-myristate 13-acetate enhanced the NMDA response. SRIF-14 permitted NMDA receptor activation in the presence of 1.2 mM Mg(2+) ions, both in hippocampal synaptosomes and slices. Blockade of inositol-1,4,5-trisphosphate (InsP(3)) receptors with heparin abolished the effect of SRIF-14. It is concluded that SRIF receptors, possibly of the sst(5) subtype, can exert a permissive role on NMDA receptors colocalized on hippocampal noradrenergic terminals: activation of sst(5) receptors is coupled to pertussis toxin-sensitive G proteins enhancing phosphoinositide metabolism with activation of InsP(3) receptors and PKC; NMDA receptor subunits might be phosphorylated with consequent removal of the Mg(2+) block in absence of depolarization. PMID- 10821784 TI - Conditions permitting suppression of stretch-induced and vasoconstrictor tone by basal nitric oxide activity in porcine cerebral artery. AB - This study examined the ability of basal nitric oxide activity to suppress intrinsic and vasoconstrictor tone in isolated rings of porcine cerebral artery. Following stretch of approximately 1 g, N(G)-nitro-L-arginine methyl ester (L NAME, 100 microM) produced a rise in tone in endothelium-containing but not endothelium-denuded rings. Thus, intrinsic tone was present and was powerfully suppressed by basal nitric oxide activity. Nevertheless, when concentration response curves were constructed to U46619 and 5-hydroxytryptamine (5-HT), no endothelium-dependent depression of vasoconstriction was observed. It therefore appeared that basal nitric oxide activity was able to suppress intrinsic but not vasoconstrictor tone in these vessels. Stretch-tension curves generated following the application of stretch over the range 0 - 5. 5 g on endothelium-denuded rings showed that tension was stretch-induced. Experiments conducted in the presence of L-NAME (100 microM) revealed that the level of tone present in endothelium containing rings was substantially higher than in endothelium-denuded rings across the entire range of stretch. When endothelium-containing and endothelium denuded rings were set at similar levels of stretch-induced tone, rather than similar levels of stretch, the presence of the endothelium now depressed significantly vasoconstrictor responses to U46619 and 5-HT. Thus, when endothelium-containing and endothelium-denuded rings of porcine cerebral artery are set at similar points along their respective stretch-tension curves, rather than at similar levels of stretch, basal nitric oxide activity can be seen to inhibit both stretch-induced and vasoconstrictor tone. PMID- 10821785 TI - Endogenous nitric oxide decreases hippocampal levels of serotonin and dopamine in vivo. AB - Nitric oxide (NO) modulates the levels of various neurotransmitters in the CNS. Here we determined whether the specific nitric oxide synthase (NOS) inhibitor 7 nitroindazole (7-NI), the non-selective inhibitor of guanylate cyclase (GC) and NOS, methylene blue (MB), the NO-precursor L-arginine (L-Arg), and the selective soluble GC inhibitor 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ) affect extracellular levels of serotonin (5-HT), dopamine (DA), 5-hydroxyindoleacetic acid (5-HIAA), and homovanillic acid (HVA) in the rat ventral hippocampus by using microdialysis in freely moving animals. Local perfusion of 7-NI (1 mM) and MB (1 mM) significantly increased extracellular level of 5-HT, whereas DA was increased by 7-NI only. Systemic administration of 7-NI (50 mg kg(-1)) and MB (30 mg kg(-1)) increased the extracellular levels of 5-HT and DA. Extracellular levels of 5-HIAA was not influenced by local or systemic MB or 7-NI. In contrast, extracellular level of HVA was decreased by systemic MB and retrodialyzed MB, but was not influenced by 7-NI. Retrodialysis of L-Arg (2 mM) decreased the levels of 5-HT, DA, 5-HIAA and HVA in the hippocampus. Systemic administration of L-Arg (250 mg kg(-1)) decreased the level of 5-HT, but failed to influence DA, 5-HIAA and HVA. Local perfusion of ODQ (400 microM) did not affect 5-HT overflow in the hippocampus. We conclude that NOS inhibitors increased extracellular levels of 5 HT and DA in the rat ventral hippocampus after local or systemic administration, whereas the NO precursor L-Arg had the opposite effect. Thus, endogenous NO may exert a negative control over the levels of 5-HT and DA in the hippocampus. However, this effect is probably not mediated by cyclic GMP. PMID- 10821786 TI - Retinoid-mediated inhibition of interleukin-12 production in mouse macrophages suppresses Th1 cytokine profile in CD4(+) T cells. AB - Interleukin-12 (IL-12) plays a central role in the immune system by driving the immune response towards T helper 1 (Th1) type responses characterized by high IFN gamma and low IL-4 production. In this study we investigated whether retinoid mediated inhibition of interleukin-12 production in mouse macrophages could regulate cytokine profile of antigen (Ag)-primed CD4(+) Th cells. Pretreatment with retinoids (9-cis-RA, all-trans-RA, TTNPB) significantly inhibited IL-12 production by mouse macrophages stimulated with lipopolysaccharide (LPS) or heated-killed Listeria monocytogenes (HKL). Retinoid-pretreated macrophages reduced their ability to induce IFN-gamma and increased the ability to induce IL 4 in Ag-primed CD4(+) T cells. Addition of recombinant IL-12 to cultures of retinoid-pretreated macrophages and CD4(+) T cells restored IFN-gamma production in CD4(+) T cells. The in vivo administration of 9-cis-RA resulted in the inhibition of IL-12 production by macrophages stimulated in vitro with either LPS or HKL, leading to the inhibition of Th1 cytokine profile (decreased IFN-gamma and increased IL-4 production) in CD4(+) T cells. These findings may explain some known effects of retinoids including the inhibition of encephalitogenicity, and point to a possible therapeutic use of retinoids in the Th1-mediated immune diseases such as autoimmune diseases. PMID- 10821787 TI - Octopamine receptors in the honey bee and locust nervous system: pharmacological similarities between homologous receptors of distantly related species. AB - Honey bees are perhaps the most versatile models to study the cellular and pharmacological basis underlying behaviours ranging from learning and memory to sociobiology. For both aspects octopamine (OA) is known to play a vital role. The neuronal octopamine receptor of the honey bee shares pharmacological similarities with the neuronal octopamine receptor of the locust. Both, agonists and antagonists known to have high affinities for the locust neuronal octopamine receptor have also high affinities for the bee neuronal octopamine receptor. The distribution of receptors is more or less congruent between locusts and bees. Optic lobes and especially the mushroom bodies are areas of greatest octopamine receptor expression in both species, which mirrors the physiological significance of octopamine in the insect nervous system. The neuronal octopamine receptor of insects served as a model to study the pharmacological similarity of homologous receptors from distantly related species, because bees and locusts are separated by at least 330 million years of evolution. PMID- 10821788 TI - Thermodynamically distinct high and low affinity states of the A(1) adenosine receptor induced by G protein coupling and guanine nucleotide ligation states of G proteins. AB - The influence of the receptor-G protein coupling state and the guanine nucleotide ligation state of the G protein on the binding mechanism of A(1) adenosine receptor ligands has been investigated in [(3)H]-1,3-dipropyl-8 cyclopentylxanthine ([(3)H]-DPCPX) binding studies in rat brain membranes. Thermodynamic parameters of binding of A(1) adenosine receptor ligands of different intrinsic activities were determined in the absence or presence of GDP and compared to the binding mechanism after receptor-G protein uncoupling. In agreement with previous studies, it was found that xanthine and non-xanthine antagonists showed an enthalpy- or enthalpy- and entropy-driven binding mechanism under all conditions. In contrast to antagonists, the binding mechanism of agonists was strongly affected by the G protein coupling state or the absence or presence of guanine nucleotides. Binding of full and partial agonists to the high affinity state of the A(1) receptor was entropy-driven in the absence of GDP, and a good correlation between intrinsic activities and the contribution of entropy was observed. In the absence of GDP, binding of full and partial agonists and antagonists to the high affinity state of the receptor was thermodynamically discriminated. In contrast, no such discrimination was found in the presence of GDP. The binding mechanism of agonists to the low-affinity state of the receptor was identical to that of antagonists only after uncoupling of the receptor from G proteins by pretreatment with N-ethylmaleimide or guanosine-5'-(gamma-thio) triphosphate (GTPgammaS). These results indicate the existence of two thermodynamically distinct high- and low-affinity states of the A(1) adenosine receptor. PMID- 10821789 TI - EDHF, NO and a prostanoid: hyperpolarization-dependent and -independent relaxation in guinea-pig arteries. AB - The contribution of endothelium-derived hyperpolarizing factor (EDHF), nitric oxide (NO) and a prostanoid (PG) to endothelium-dependent hyperpolarization and relaxation were assessed in coronary and mammary arteries of guinea-pigs by integration of the responses evoked during discrete applications of acetylcholine (ACh). The results of this integration approach were compared with those using traditional peak analysis methods. N(omega)-nitro-L-arginine methyl ester (L NAME, 100 microM) and indomethacin (1 microM), alone or in combination, were without effect on peak hyperpolarizations or relaxations while they markedly reduced the integrated responses in both arteries. Integrated responses attributed to NO and PG were larger than those attributed to EDHF in the coronary artery (at 2 microM ACh, hyperpolarization (mV s): NO, 4200+/-91; PG, 5046+/-157; EDHF, 1532+/-94; relaxation (mN s mm(-1)): NO, 2488+/-122; PG, 2234+/-96; EDHF, 802+/-54). Integrated responses attributed to NO, PG and EDHF were similar in the mammary artery (at 2 microM ACh, hyperpolarization: NO, 347+/-69; PG, 217+/-49; EDHF, 310+/-63; relaxation: NO, 462+/-94; PG, 456+/-144; EDHF, 458+/-40). Gilbenclamide (1 microM) all but abolished the hyperpolarization attributable to NO and PG but not EDHF in both arteries allowing assessment of the role of the hyperpolarization in relaxation. Gilbenclamide was without effect on the integrated relaxation due to NO but significantly reduced the relaxation associated with PG in the two arteries. In conclusion, integration of the responses enabled a more complete assessment of the contribution of EDHF, NO and PG to endothelium-dependent responses, which were strikingly different in the two arteries. There is commonality in the role of hyperpolarization in relaxation in both arteries: EDHF-dependent relaxation is strongly dependent on hyperpolarization; hyperpolarization plays an important role in PG relaxation, whereas it has a small facilitatory role in NO-dependent relaxation. PMID- 10821790 TI - Effects of halothane on the membrane potential in skeletal muscle of the frog. AB - Halothane has many effects on the resting membrane potential (V(m)) of excitable cells and exerts numerous effects on skeletal muscle one of which is the enhancement of Ca(2+) release by the sarcoplasmic reticulum (SR) resulting in a sustained contracture. The aim of this study was to analyse the effects of clinical doses of halothane on V(m), recorded using intracellular microelectrodes on cleaned and non stimulated sartorius muscle which was freshly isolated from the leg of the frog Rana esculenta. We assessed the mechanism of effects of superfused halothane on V(m) by the administration of selective antagonists of membrane bound Na(+), K(+) and Cl(-) channels and by inhibition of SR Ca(2+) release. Halothane (3%) induced an early and transient depolarization (4.5 mV within 7 min) and a delayed and sustained hyperpolarization (about 11 mV within 15 min) of V(m). The halothane-induced transient depolarization was sensitive to ryanodine (10 microM) and to 4-acetamido-4'-isothiocyanatostilbene 2,2' disulphonic acid (SITS, 1 mM). The hyperpolarization of V(m) induced by halothane (0.1 - 3%) was dose-dependent and reversible. It was insensitive to SITS (1 mM), tetrodotoxin (0.6 microM), and tetraethylammonium (10 mM) but was blocked and/or prevented by ryanodine (10 microM), charybdotoxin (CTX, 1 microM), and glibenclamide (10 nM). Our observations revealed that the effects of halothane on V(m) may be related to the increase in intracellular Ca(2+) concentration produced by the ryanodine-sensitive Ca(2+) release from the SR induced by the anaesthetic. The depolarization may be attributed to the activation of Ca(2+) dependent Cl(-) (blocked by SITS) channels and the hyperpolarization to the activation of large conductance Ca(2+)-dependent K(+) channels, blocked by CTX, and to the opening of ATP-sensitive K(+) channels, inhibited by glibenclamide. PMID- 10821791 TI - Properties and pharmacological modification of ATP-sensitive K(+) channels in cat tracheal myocytes. AB - The effects of levcromakalim and nucleoside diphosphates (NDPs) on both membrane currents and unitary currents in cat trachea myocytes were investigated by use of patch-clamp techniques. In conventional whole-cell configuration, levcromakalim produced a concentration-dependent K(+) current which was suppressed by additional application of 5 microM glibenclamide at -70 mV. When 3 mM ATP was added in the pipette solution, the peak amplitude of the levcromakalim-induced current was much smaller than that in the absence of ATP. When 3 mM uridine 5' diphosphate (UDP) was included in the pipette solution, much higher concentrations of glibenclamide (>/=50 microM) were required to suppress the 100 microM levcromakalim-induced membrane current in comparison with those in the absence of UDP. In the cell-attached patches, levcromakalim activated a 40 pS K(+) channel which was inhibited by additional application of glibenclamide in symmetrical 140 mM K(+) conditions. UDP (>/=0.1 mM) was capable of reactivating the channel in inside-out patches in which the glibenclamide-sensitive K(+) channel had run down, in the presence of levcromakalim. The K(+) channel reactivated by UDP was suppressed by additional application of either intracellular 3 mM ATP or 100 microM glibenclamide. These results demonstrate that smooth muscle cells in the cat trachea possess ATP-sensitive 40 pS K(+) channels which are blocked by glibenclamide (i.e. K(ATP)) and can be activated by levcromakalim and that intracellular UDP causes a significant shift of the glibenclamide-sensitivity of these channels. PMID- 10821792 TI - The mechanism of cryoinjury: In vitro studies on human internal mammary arteries. AB - The mechanism of cryoinjury was investigated in human internal mammary arteries (IMA) by monitoring contractile responses to ET-1 and KCl. For cryopreservation segments of IMA were equilibrated for 20 min with the cryomedium (RPMI 1640 culture medium containing 1.8 M DMSO and 0.1 M sucrose), frozen at a mean cooling rate of 1.3 degrees C min(-1) to -70 degrees C and stored in liquid nitrogen. Before use, samples were thawed slowly and the cryomedium removed by dilution. Compared to unfrozen controls, ET-1 stimulated frozen/thawed IMA with similar efficacy but at 3 fold lower concentrations (P<0.001). Addition of ET-1 (100 nM) induced maximal contraction of unfrozen IMA within 10 min, declining thereafter to 25% after 90 min. In frozen/thawed IMA the ET-1-induced contraction was sustained but could be reversed if protein kinase C was blocked by staurosporine (100 nM). Responses to ET-1 of cryostored IMA were 5 fold more susceptible to blockade by nifedipine than those of controls. After cryostorage the efficacy of KCl was diminished to 40% (P<0.05) and the KCl curve was shifted to the left (2 fold, P<0. 001). In both unfrozen and cryostored IMA the KCl (60 mM) effect was sustained and equally susceptible to nifedipine. It is suggested that the smooth muscle cell of IMA is receptive to physical forces which occur during cryopreservation. These forces modify transmembrane signal transduction and intracellular pathways, that are common to pharmacological agonists thereby changing vascular responses to several contractile agonists after thawing. PMID- 10821793 TI - Cyclo-oxygenase-2 enhances basic fibroblast growth factor-induced angiogenesis through induction of vascular endothelial growth factor in rat sponge implants. AB - Angiogenesis is reportedly enhanced by prostaglandins (PGs). In the present study, we investigated whether or not cyclo-oxygenase (COX)-2 mediated angiogenesis in chronic and proliferate granuloma. In rat sponge implants, angiogenesis was gradually developed over a 14-day experimental period as granuloma formed. This angiogenesis was enhanced by topical injections of human recombinant basic fibroblast growth factor (bFGF). In sponge granuloma, mRNA of COX-1 was constitutively expressed, whereas that of COX-2 was increased with neovascularization in parallel with that of vascular endothelial growth factor (VEGF). Topical injections of bFGF increased the expression of COX-2 mRNA. bFGF stimulated angiogenesis was inhibited by indomethacin or selective COX-2 inhibitors, NS-398, nimesulide, and JTE-522. The levels of PGE(2) and 6-keto PGF(1alpha) in the sponge granuloma were increased with bFGF 13 fold and 9 fold, respectively, and these levels were markedly reduced by NS-398. The expression of VEGF mRNA in the granuloma was also enhanced by bFGF, and was reduced by NS-398. Topical injections of PGE(2) and beraprost sodium, a PGI(2) analogue, increased the expression of VEGF mRNA, with angiogenesis enhancement. The enhanced angiogenesis by bFGF was significantly inhibited by topical injections of VEGF anti-sense oligonucleotide. These results suggested that COX-2 may enhance bFGF induced neovascularization in sponge granuloma by PG-mediated expression of VEGF, and that a COX-2 inhibitor would facilitate the management of conditions involving angiogenesis. PMID- 10821794 TI - Xestospongin C, a selective and membrane-permeable inhibitor of IP(3) receptor, attenuates the positive inotropic effect of alpha-adrenergic stimulation in guinea-pig papillary muscle. AB - We evaluated the role of the inositol 1,4,5-triphosphate (IP(3)) receptor mediated Ca(2+) release on the positive inotropic effects of alpha-adrenergic stimulation using a novel, potent, selective membrane-permeable blocker of IP(3) receptor, xestospongin C. Guinea-pig papillary muscle permeabilized with saponin exhibited spontaneous oscillatory contractions in solution buffered with pCa(2+) 6.5 by a low concentration of EGTA. The oscillatory activity was increased by adding 100 microM IP(3) and abolished by 1 microM ryanodine or 30 microM cyclopiazonic acid. Xestospongin C (3 microM) inhibited the IP(3)-induced increase in the oscillatory contractions without affecting basal oscillations. In intact papillary muscle, xestospongin C (3 microM) inhibited the positive inotropic effects of phenylephrine, resulting in a rightward and downward shift of the concentration-response curve for phenylephrine. On the contrary, xestospongin C did not affect the concentration-response curve for phenylephrine obtained in the presence of ryanodine (1 microM). On the other hand, xestospongin C affected neither basal contractions nor the positive inotropic effects of a high extracellular Ca(2+) concentration (3.2 mM) or that of isoprenaline (1 and 10 nM). These results suggest that the IP(3)-mediated increase in Ca(2+) release is involved in the positive inotropic effects of alpha-adrenergic stimulation in the guinea-pig cardiac muscle. PMID- 10821795 TI - [(3)H]-trimetazidine mitochondrial binding sites: regulation by cations, effect of trimetazidine derivatives and other agents and interaction with an endogenous substance. AB - Trimetazidine, an antiischaemic drug, has been shown to restore impaired mitochondrial functions. Specific binding sites for [(3)H]-trimetazidine have been previously detected in liver mitochondria. In the present study we confirm this observation and provide additional evidence for the involvement of these sites in the pharmacological effects of the drug. Inhibition experiments using a series of trimetazidine derivatives revealed the presence of three classes of binding sites. An N-benzyl substituted analogue of trimetazidine exhibited a very high affinity (K(i)=7 nM) for one of these classes of sites. Compounds from different pharmacological classes were evaluated for their ability to inhibit [(3)H]-trimetazidine binding. Among the drugs tested pentazocine, ifenprodil, opipramol, perphenazine, haloperidol, and to a lower extent prenylamine, carbetapentane and dextromethorphan competed with high affinity, suggesting a similarity of high affinity [(3)H]-trimetazidine sites with sigma receptors. [(3)H]-Trimetazidine binding was modulated by pH. Neutral trimetazidine had about 10 fold higher affinity than protonated trimetazidine for its mitochondrial binding sites. Various cations also affected [(3)H]-trimetazidine binding. Ca(2+) was the most potent inhibitor and totally suppressed the binding of [(3)H] trimetazidine to the sites of medium affinity. An endogenous cytosolic ligand was able to displace [(3)H]-trimetazidine from its binding sites. Its activity was not affected by boiling for 15 min, suggesting a non-protein compound. These data suggest that mitochondrial [(3)H]-trimetazidine binding sites could have a physiological relevance and be involved in the antiischaemic effects of the drug. PMID- 10821796 TI - Evidence that 2-methylthioATP and 2-methylthioADP are both agonists at the rat hepatocyte P2Y(1) receptor. AB - In the absence of selective antagonists, pharmacological characterization of P2Y receptor subtypes has relied heavily upon their distinct agonist profiles. 2 methylthioADP (2-MeSADP) is a selective agonist for the P2Y(1) receptor. The agonist action of 2-MeSATP at the P2Y(1) receptor has recently been questioned. The effects of both 2-MeSADP and 2-MeSATP have been studied on rat hepatocytes injected with the bioluminescent Ca(2+) indicator, aequorin. Single hepatocytes generate series of repetitive transients in cytosolic free calcium concentration ([Ca(2+)](i)) when stimulated with agonists acting through the phosphoinositide signalling pathway. The transients induced by 2-MeSADP and 2-MeSATP in the same cell were indistinguishable, indicating that they act at a common receptor. In contrast the transients evoked by ATP and UTP had very different profiles. Treatment of 2-MeSATP with an ATP-regenerating system to remove contaminating 2 MeSADP did not abolish its agonist activity. Application of the P2Y(1) antagonist, adenosine-3'-phosphate-5'-phosphate (A3P5P) inhibited the transients induced by both 2-MeSADP and 2-MeSATP. In contrast the transients induced by ATP and UTP were enhanced by the addition of A3P5P. These results indicate that both 2-MeSADP and 2-MeSATP are agonists at the rat hepatocyte P2Y(1) receptor. PMID- 10821797 TI - High affinity interaction of mibefradil with voltage-gated calcium and sodium channels. AB - Mibefradil is a novel Ca(2+) antagonist which blocks both high-voltage activated and low voltage-activated Ca(2+) channels. Although L-type Ca(2+) channel block was demonstrated in functional experiments its molecular interaction with the channel has not yet been studied. We therefore investigated the binding of [(3)H] mibefradil and a series of mibefradil analogues to L-type Ca(2+) channels in different tissues. [(3)H]-Mibefradil labelled a single class of high affinity sites on skeletal muscle L-type Ca(2+) channels (K(D) of 2.5+/-0.4 nM, B(max)=56.4+/-2.3 pmol mg(-1) of protein). Mibefradil (and a series of analogues) partially inhibited (+)-[(3)H]-isradipine binding to skeletal muscle membranes but stimulated binding to brain L-type Ca(2+) channels and alpha1C-subunits expressed in tsA201 cells indicating a tissue-specific, non-competitive interaction between the dihydropyridine and mibefradil binding domain. [(3)H] Mibefradil also labelled a heterogenous population of high affinity sites in rabbit brain which was inhibited by a series of nonspecific Ca(2+) and Na(+) channel blockers. Mibefradil and its analogue RO40-6040 had high affinity for neuronal voltage-gated Na(+)-channels as confirmed in binding (apparent K(i) values of 17 and 1.0 nM, respectively) and functional experiments (40% use dependent inhibition of Na(+)-channel current by 1 microM mibefradil in GH3 cells). Our data demonstrate that mibefradil binds to voltage-gated L-type Ca(2+) channels with very high affinity and is also a potent blocker of voltage-gated neuronal Na(+)-channels. More lipophilic mibefradil analogues may possess neuroprotective properties like other nonselective Ca(2+)-/Na(+)-channel blockers. PMID- 10821798 TI - Selective inhibitory effects of niflumic acid on 5-HT-induced contraction of the rat isolated stomach fundus. AB - The effects of niflumic acid (NFA), an inhibitor of calcium-activated chloride currents I(Cl(Ca)), were compared with the actions of the voltage-dependent calcium channel (VDCC) blocker nifedipine on 5-hydroxtryptamine (5-HT)- and acetylcholine (ACh)-induced contractions of the rat isolated fundus. NFA (1 - 30 microM) elicited a concentration-dependent inhibition of contractions induced by 5-HT (10 microM) with a reduction to 15. 5+/-6.0% of the control value at 30 microM. 1 microM nifedipine reduced 5-HT-induced contraction to 15.2+/-4.9% of the control, an effect not greater in the additional presence of 30 microM NFA. In contrast, the contractile response to ACh (10 microM) was not inhibited by NFA in concentrations /=10 microM. Our results show that NFA can exert selective inhibitory effects on the chloride-dependent 5-HT-induced contractions of the rat fundus. The data support the hypothesis that activation of Cl((Ca)) channels leading to calcium entry via VDCCs is a mechanism utilized by 5-HT, but not by ACh, to elicit contraction of the rat fundus. PMID- 10821799 TI - A(2A) adenosine receptor mediated potassium channel activation in rat epididymal smooth muscle. AB - The effects of A(2) adenosine receptor agonists upon phenylephrine-stimulated contractility in preparations of rat epididymis were investigated. Preparations responded to phenylephrine (3 microM) with submaximal contractions. Adenosine and the stable agonists 5'-N-ethylcarboxamido-adenosine (NECA) and 2-p-(2 carboxyethyl) phenethylamino-N-ethylcarboxamide adenosine (CGS 21680) inhibited phenylephrine-induced contractions (potency order, NECA>CGS 21680>adenosine). The A(2A) receptor-selective antagonist, 4-(2-[7-amino-2-(2-furyl)[1,2,4]-triazolo [2,3-a][1,3, 5]triazin-5-ylamino]ethyl)phenol (ZM 241385, 30 microM) blocked the response to NECA. The A(2A) adenosine receptor-mediated inhibitory responses to NECA were reduced by the K(ATP) channel blocker, glibenclamide (3 microM) and abolished by charybdotoxin (100 nM). The diterpene forskolin elicited a concentration-dependent inhibition of phenylephrine (3 microM)-stimulated contractility (by 62+/-8% of control at 100 microM). Charybdotoxin (100 nM), but not glibenclamide (3 microM) blocked the forskolin (10 microM) inhibition of phenylephrine-stimulated contractility. NECA elicited concentration-dependent increases in both cyclic AMP and cyclic GMP accumulation which were antagonized by ZM 241385 (30 nM). The protein kinase G activator, APT-cyclic GMP (8-( Aminophenylthio) guanosine-3',5'-cyclic monophosphate) and the protein kinase A activator (Sp)-8-bromoadenosine-3',5'-cyclic monophosphorothioate (Sp-8-Br-cyclic AMPs), inhibited phenylephrine (3 microM) induced contractions of rat epididymis. Glibenclamide (3 microM), but not charybdotoxin (100 nM), inhibited ATP-cyclic GMP responses. Charybdotoxin (100 nM), but not glibenclamide (3 microM) reduced the effect of Sp-8-Br-cyclic AMPs. This study shows that the A(2A) adenosine receptor inhibition of epididymal contractility may be mediated through the activation of charybdotoxin- and glibenclamide-sensitive potassium channels and may involve the activation of both protein kinases A and G. PMID- 10821800 TI - Further evidence that 5-HT-induced relaxation of pig pulmonary artery is mediated by endothelial 5-HT(2B) receptors. AB - The endothelial 5-HT receptor mediating relaxation of pig pulmonary artery has been characterized using the selective 5-HT(2B) receptor agonist BW 723C86 and a variety of structurally diverse 5-HT receptor antagonists. If arterial rings with intact endothelium were precontracted with prostaglandin F(2alpha) (3 microM), BW 723C86 caused concentration-dependent relaxation with a pEC(50)=8.21+/-0.03 and E(max)=89+/-4% relative to 5-HT. The relaxant responses to BW 723C86 were inhibited by the 5-HT(2B) receptor antagonist SB 204741, the 5-HT(2B/2C) receptor antagonist SB 206553 and the antimigraine drug pizotifen, yielding pA(2) values of 6.68, 7.20 and 8.32, respectively. The pA(2) values against BW 723C86 were similar to those determined against 5-HT. The relaxant effect of 5-HT was antagonized by a variety of 22 compounds of diverse chemical structures. Based on the calculated mean pA(2) values the order of the most potent antagonists was ritanserin (9.38) > methysergide (8. 86) > pizotifen (8.47) >/= methiothepin (8.32) > LY 53857 (7.84) >/= amoxapine (7.80) >/= loxapine (7.73) >/= metergoline (7.64) >/= mianserin (7.51) >/= rauwolscine (7.39). Compounds with weak blocking potency were yohimbine (6.37), spiperone (5.88) and ketanserin (5.85). Correlation analysis between the affinities of the antagonists in pig pulmonary artery and those from radioligand binding studies at human and rat 5-HT(2B) receptors showed a highly significant correlation (r=0.95 and 0.84, P<0.002 and <0.005). Correlation with 5-HT(2C) receptors was much lower (r=0.57, P=0.035), and no correlations were obtained with 5-ht(6) and 5-HT(7) receptors. It is concluded that the 5-HT receptor mediating endothelium-dependent relaxation of pig pulmonary artery is of the 5-HT(2B) subtype. PMID- 10821801 TI - Long-lasting cholecystokinin(2) receptor blockade after a single subcutaneous injection of YF476 or YM022. AB - Histamine-forming ECL cells in the rat stomach operate under the control of gastrin. They represent a convenient target for studying cholecystokinin B/gastrin (CCK(2)) receptor antagonists in vivo. We examined the effectiveness and duration of action of two CCK(2) antagonists, YM022 and YF476, with respect to their effect on ECL-cell histidine decarboxylase (HDC) activity in the rat. Oral administration of subcutaneous deposition of YF476 or YM022 reduced the HDC activity. The maximum/near-maximum dose for both drugs and for both modes of administration was 300 micromol kg(-1) (effects measured 24 h after dose). At this dose and time the serum concentration of YF476 was 20 - 40 nmol l(-1). The dose 300 micromol kg(-1) was used in all subsequent studies. A single subcutaneous injection of YF476 inhibited the HDC activity for 8 weeks. The circulating concentration of YF476 remained high for the same period of time (>/=15 nmol l(-1)). Subcutaneous YM022 suppressed the HDC activity for 4 weeks. A single oral dose of YF476 or YM022 inhibited the HDC activity for 2 - 3 days. Chronic gastric fistula rats were used to study the effect of subcutaneous YF476 on gastrin-stimulated acid secretion. A single injection of YF476 prevented gastrin from causing an acid response for at least 4 weeks (the longest time studied). We conclude that a single subcutaneous injection of 300 micromol kg(-1) YF476 causes blockade of CCK(2) receptors in the stomach of the rat for 8 weeks thus providing a convenient method for studies of the consequences of long-term CCK(2) receptor inhibition. PMID- 10821803 TI - May 23, 2000 PMID- 10821802 TI - Increased defecation during stress or after 5-hydroxytryptophan: selective inhibition by the 5-HT(4) receptor antagonist, SB-207266. AB - 5-HT(4) receptor antagonism prevents the ability of exogenous 5-HT or 5-HTP to sensitize the intestinal peristaltic reflex and increase the rate of defecation, generally without affecting non-stimulated intestinal function. In this study we confirmed the ability of the selective 5-HT(4) receptor antagonist SB-207266 1 - 1000 microg kg(-1) p.o., to prevent the increase in defecation evoked over a 60 min period by 5-HTP 10 mg kg(-1) s.c. in conscious mice, in the absence of an apparent constipating action. The role of endogenous 5-HT in the mechanisms of increased defecation and/or diarrhoea was then investigated in conscious, fed rats. This was evoked by 180 min exposure to restraint stress, which increased both the number and mean weight of formed, faecal pellets excreted over the entire time period. SB-207266 1 - 1000 microg kg(-1) p.o. (dosed 30 min before restraint) did not affect the increase in defecation evoked during the first 60 min of restraint stress, but significantly and dose-dependently reduced or prevented the increased defecation during the remaining 120 min of the experiment; this action occurred in the absence of an apparent constipating action of SB-207266. In fasted rats exposed to restraint stress, watery diarrhoea developed and although there was a tendency for SB-207266 1 - 1000 microg kg(-1) p.o. (dosed 30 min before restraint) to reduce the incidence of diarrhoea, this inhibition was not complete. We conclude that selective 5-HT(4) receptor antagonism prevents disruptions in defecation behaviours caused by exogenous or endogenous enteric 5-HT and that this activity is not accompanied by a concomitant suppression of activity (constipation-like) within the intestine itself. PMID- 10821804 TI - Beyond TIMI III flow. PMID- 10821805 TI - Angiotensin-1 receptor autoantibodies: A role in the pathogenesis of preeclampsia? PMID- 10821806 TI - Chronic beta-adrenergic stimulation induces myocardial proinflammatory cytokine expression. AB - BACKGROUND: The sympathetic nervous system and proinflammatory cytokines are believed to play key roles in the pathophysiology of congestive heart failure. To evaluate a possible relationship between these neurohormonal systems, we studied the effects of chronic beta-adrenergic stimulation on the myocardial and systemic elaboration of tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, and IL 6. METHODS AND RESULTS: Male rats received either L-isoproterenol (2.4 mg. kg( 1). d(-1), n=8) or saline (n=7) via miniosmotic pumps for 7 days. Myocardial cytokine expression was analyzed by both Northern and Western blotting and localized in the tissue using immunohistochemistry. ELISA was performed to measure circulating levels of cytokines. In myocardium from control animals, neither TNF-alpha nor IL-1beta were detected, whereas IL-6 was present at very low levels. Isoproterenol led to a significant (P<0.01) increase in mRNA and protein expression of all 3 cytokines. Immunohistochemistry did not detect immunoreactivity for either cytokine in myocardium from controls; however, all 3 cytokines were readily detected (P<0.05) throughout the myocardium, localized to resident cells and vessels, in animals treated with isoproterenol. Neither treatment group had detectable levels of cytokines in the serum. CONCLUSIONS: Chronic beta-adrenergic stimulation induces myocardial, but not systemic, elaboration of TNF-alpha, IL-1beta, and IL-6. PMID- 10821807 TI - Estrogen-induced vasoprotection is estrogen receptor dependent: evidence from the balloon-injured rat carotid artery model. AB - BACKGROUND: Previous studies have shown that estrogen (E2) is vasoprotective in multiple animal models of vascular injury, including mice with homologous disruptions of either the alpha or beta isoforms of the estrogen receptor (ER) gene, calling into question the ER dependency of the vasoprotective effect. This study used ICI 182,780, a nonselective ER antagonist, to test the hypothesis that the vasoprotective effect of E2 in the rat carotid injury model is ER mediated. METHODS AND RESULTS: Intact female Sprague-Dawley rats were divided into 4 groups and treated with the nonselective ER antagonist ICI 182,780 (ICI; 0.5, 1.5, or 5 mg. kg(-1). d(-1), subcutaneously [S.C.]) or vehicle, beginning before balloon injury of the right common carotid artery and continuing for 14 days afterward. Four groups of ovariectomized rats (OVX) were treated with 17beta estradiol (E2) (20 microgram. kg(-1). d(-1), S.C.) alone or combined with ICI 5 mg. kg(-1). d( 1), S.C.; with ICI 5 mg. kg(-1). d(-1) alone; or with vehicle according to a similar protocol. Two weeks after injury, rats were killed, and the carotid arteries were evaluated for neointima formation using morphometric analysis. ICI 182,780 blunted the E2-related protective effect and increased neointima formation in injured carotid arteries of intact female rats in a dose-dependent fashion. ICI had no effect on neointima formation in OVX, but addition of ICI to E2 in OVX blocked the inhibitory effect of exogenous E2 on neointima formation. CONCLUSIONS: These results indicate that the vasoprotective effect of E2 in the balloon-injured rat carotid artery model is mediated by ER. PMID- 10821808 TI - Abnormal aortic valve development in mice lacking endothelial nitric oxide synthase. AB - BACKGROUND: Endothelium-derived nitric oxide (NO) is produced by an oxidative reaction catalyzed by endothelial NO synthase (eNOS). NO plays a crucial role in controlling cell growth and apoptosis, as well as having well-characterized vasodilator and antithrombotic actions. More recently, endothelium-derived NO was shown to be involved in postdevelopmental vascular remodeling and angiogenesis, as well as in the formation of limb vasculature during embryogenesis. Therefore, we investigated the role of endothelium-derived NO during cardiovascular development using mice deficient in eNOS. METHODS AND RESULTS: We examined the hearts of 12 mature eNOS-deficient and 26 mature wild-type mice. Five of the mature eNOS-deficient mice had a bicuspid aortic valve; none of the 26 wild-type animals exhibited identifiable valvular or cardiac abnormalities. Immunohistochemical analysis revealed prominent eNOS expression localized to the endothelium lining the valve cusps of the aorta in mature wild-type mice; expression was localized to the myocardium and endothelial cell monolayer lining the valve leaflets in the developing embryo. CONCLUSIONS: These results show a strong association between eNOS deficiency and the presence of a bicuspid aortic valve; they provide the first molecular insight into one of the most common types of congenital cardiac abnormality. PMID- 10821809 TI - Acute and chronic angiotensin-1 receptor antagonism reverses endothelial dysfunction in atherosclerosis. AB - BACKGROUND: The renin-angiotensin system may contribute to atherogenesis through the promotion of endothelial dysfunction. The present study was performed to determine whether angiotensin-1 (AT(1)) receptor inhibition improves endothelial dysfunction. METHODS AND RESULTS: In the femoral circulation of 19 patients with atherosclerosis and of 9 control subjects, we studied microvascular responses to reactive hyperemia, angiotensin II, acetylcholine, and sodium nitroprusside before and after the administration of intra-arterial losartan (10 mg). Femoral artery flow velocity was measured with a Doppler flow wire, and the femoral vascular resistance index (FVRI) was calculated as mean arterial pressure divided by flow velocity. Losartan induced a minor (5.9+/-2%, P=0. 02) reduction in FVRI and inhibited angiotensin II-mediated vasoconstriction in both patient groups (P<0.01). After the administration of losartan, acetylcholine-mediated vasodilation was augmented in patients (44+/-5% to 58+/-4% reduction in FVRI with infusion at a rate of 150 microgram/min, P<0.001) but not control subjects. Vasodilation during reactive hyperemia was also greater after AT(1) receptor inhibition (P=0.03) in patients, but the response to sodium nitroprusside remained unchanged. In a separate group of 31 patients with atherosclerosis, we investigated the effect of 8 weeks of oral losartan therapy on brachial artery flow-mediated vasodilation with the use of high-resolution ultrasound. Oral losartan therapy improved flow-mediated brachial artery dilation (1.4+/-0.9% to 3.2+/-0.8%, P=0.03) but had no effect on the nitroglycerin response. Serum nitrogen oxide levels increased from 21.6+/-1.7 to 26.7+/-2.4 micromol/L (P=0.008). CONCLUSIONS: The results of the present study indicate that inhibition of the AT(1) receptor in patients with atherosclerosis reverses endothelial dysfunction by improving NO availability and therefore may have long-term therapeutic benefits. PMID- 10821810 TI - Intracoronary beta-irradiation with a liquid (188)re-filled balloon: six-month results from a clinical safety and feasibility study. AB - BACKGROUND: Coronary irradiation is a new concept to reduce restenosis. We evaluated the feasibility and safety of intracoronary irradiation with a balloon catheter filled with (188)Re, a liquid, high-energy beta-emitter. METHODS AND RESULTS: Irradiation with 15 Gy at 0.5-mm tissue depth was performed in 28 lesions after balloon dilation (n=9) or stenting (n=19). Lesions included 19 de novo stenoses, 4 occlusions, and 5 restenoses. Irradiation time was 515+/-199 seconds in 1 to 4 fractions. There were no procedural complications. One patient died of noncardiac causes at day 23. One asymptomatic patient refused 6-month angiography. Quantitative angiography after intervention showed a reference diameter of 2. 77+/-0.35 mm and a minimal lumen diameter of 2.36+/-0.43 mm. At 6 month follow-up, minimal lumen diameter was 1.45+/-0.88 mm (late loss index 0.57). Target lesion restenosis rate (>50% in diameter) was low (12%; 3 of 26). In addition, we observed 9 stenoses at the proximal or distal end of the irradiation zone, potentially caused by the short irradiation segment and the decreasing irradiation dose at its borders ("edge" stenoses). The total restenosis rate was 46% and was significantly lower (29% vs 70%, P=0.042) when the length of the irradiated segment was more than twice the lesion length. CONCLUSIONS: Coronary irradiation with a (188)Re-filled balloon is technically feasible and safe, requiring only standard percutaneous transluminal coronary angioplasty techniques. The target lesion restenosis rate was low. The observed edge stenoses appear to be avoidable by increasing the length of the irradiated segment. PMID- 10821811 TI - Relation of phasic coronary flow velocity characteristics with TIMI perfusion grade and myocardial recovery after primary percutaneous transluminal coronary angioplasty and rescue stenting. AB - BACKGROUND: A residual stenosis and/or microvascular damage have been proposed as mechanisms of TIMI 2 flow for acute myocardial infarction. Coronary flow dynamics were assessed in patients with TIMI 2 flow to predict whether additional intervention would improve TIMI grade. METHODS AND RESULTS: In 35 patients who had a successfully recanalized anterior acute myocardial infarction using angioplasty or rescue stenting, coronary flow patterns were compared with corresponding TIMI grade and regional left ventricular wall motion (LVWM) 1 month after the intervention. After angioplasty, the time-averaged peak velocity (APV) was lower in patients with TIMI 2 flow (n=22) than in those with TIMI 3 flow (n=13; 7.9+/-3.9 versus 20.6+/-5.1 cm/s; P<0.001). Two different flow patterns were recorded in patients with TIMI 2 flow (versus TIMI 3, P<0.001); patients with type 1 TIMI 2 flow (n=15) had a reduced diastolic APV (8.3+/-4.8 versus 24.2+/-7.4 cm/s), prolonged diastolic deceleration time (1176+/-455 versus 728+/ 205 ms), and a small diastolic/systolic APV ratio (1.3+/-0.6 versus 2.1+/-0.7); patients with type 2 TIMI 2 flow (n=7) had systolic flow reversal (systolic APV, 7.9+/-4.6 versus 11. 7+/-4.5 cm/s), a rapid diastolic deceleration time (221+/-84 versus 728+/-205 ms), and a negative diastolic/systolic APV ratio (-2.1+/-1. 4 versus 2.1+/-0.7). A significantly lower mean chord LVWM (-3.0+/-0. 2 versus 1.9+/-0.8; P<0.001) and a greater number of chords <-2SD (50+/-2 versus 28+/-18; P<0.001) were present in patients with type 2 versus type 1 TIMI 2 flow. Stenting increased TIMI 2 flow to TIMI 3 flow more in patients with type 1 than type 2 flow (67% versus 0%; P=0.003). Patients with TIMI 2 flow after stenting continued to demonstrate a type 2 pattern, and they had poor LVWM recovery. CONCLUSIONS: The differentiation between 2 types of TIMI 2 flow can predict the improvement of TIMI grade and LVWM recovery after additional stenting. PMID- 10821812 TI - Assessment of myocardial reperfusion by intravenous myocardial contrast echocardiography and coronary flow reserve after primary percutaneous transluminal coronary angioplasty [correction of angiography] in patients with acute myocardial infarction. AB - BACKGROUND: This study investigated whether the extent of perfusion defect determined by intravenous myocardial contrast echocardiography (MCE) in patients with acute myocardial infarction (AMI) treated by primary percutaneous transluminal coronary angioplasty (PTCA) relates to coronary flow reserve (CRF) for assessment of myocardial reperfusion and is predictive for left ventricular recovery. METHODS AND RESULTS: Twenty-five patients with first AMI underwent intravenous MCE with NC100100 with intermittent harmonic imaging before PTCA and after 24 hours. MCE before PTCA defined the risk region and MCE at 24 hours the "no-reflow" region. The no-reflow region divided by the risk region determined the ratio to the risk region. CFR was assessed immediately after PTCA and 24 hours later. Left ventricular wall motion score indexes were calculated before PTCA and after 4 weeks. CFR at 24 hours defined a recovery (CFR >/=1.6; n=17) and a nonrecovery group (CFR <1.6; n=8). Baseline CFR did not differ between groups. MCE ratio to the risk region was smaller in the recovery group compared with the nonrecovery group (34+/-49% vs 81+/-46%, P=0.009). A ratio to the risk region of 3 months after successful percutaneous intervention underwent PC-MRI measurements of coronary artery flow reserve followed by assessments of stenosis severity with computer-assisted quantitative coronary angiography. The intervention was performed in the left anterior descending coronary artery in 15 patients, one of its diagonal branches in 2 patients, and the right coronary artery in 1 patient. A PC-MRI coronary flow reserve value /=70% and >/=50%, respectively. CONCLUSIONS: Assessments of coronary flow reserve with PC-MRI can be used to identify flow-limiting stenoses (luminal diameter narrowings >70%) in patients with recurrent chest pain in the months after a successful percutaneous intervention. PMID- 10821815 TI - Effects of iloprost inhalation on exercise capacity and ventilatory efficiency in patients with primary pulmonary hypertension. AB - BACKGROUND: The continuous infusion of prostacyclin has been shown to improve exercise capacity and survival in patients with primary pulmonary hypertension (PPH). Inhalation of iloprost, a stable analog of prostacyclin, might be an alternative therapy for PPH, selectively acting on the pulmonary vascular bed through ventilation-matched alveolar deposition of the drug. We investigated the short-term effects of iloprost inhalation on exercise capacity and gas exchange in patients with PPH. METHODS AND RESULTS: In 11 patients with PPH, we performed 2 consecutive cardiopulmonary exercise tests before and after the inhalation of 17 microgram of iloprost. Patients had marked pulmonary hypertension (mean pulmonary artery pressure 65 mm Hg), and inhalation resulted in a decrease in pulmonary vascular resistance (1509 versus 1175 dyne. s(-1). cm(-5), P<0.05). Arterial blood gases remained unchanged (PaO(2) 69.3 versus 66.8 mm Hg; PaCO(2) 29.6 versus 28.8 mm Hg). Iloprost significantly (P<0.05) improved exercise duration (379 versus 438 seconds), peak oxygen uptake (12.8 versus 14.2 mL. kg( 1). min(-1)), VE-versus-V CO(2) slope (58 versus 51.4). CONCLUSIONS: The present data show that iloprost inhalation exerts pulmonary vasodilatation and improves symptoms and exercise capacity in patients with PPH. The data also suggest that iloprost inhalation is a suitable treatment for PPH. Whether these effects are maintained during long-term treatment and are paralleled by improvement in prognosis remains to be determined. PMID- 10821814 TI - AT(1) receptor agonistic antibodies from preeclamptic patients cause vascular cells to express tissue factor. AB - BACKGROUND: We recently described autoantibodies (angiotensin-1 receptor autoantibodies, AT(1)-AA) directed at the AT(1) receptor in the serum of preeclamptic patients, whose placentas are commonly infarcted and express tissue factor (TF). Mechanisms of how AT(1)-AA might contribute to preeclampsia are unknown. We tested the hypothesis that AT(1)-AA cause vascular smooth muscle cells (VSMC) to express TF. METHODS AND RESULTS: IgG from preeclamptic patients containing AT(1)-AA was purified with anti-human IgG columns. AT(1)-AA were separated from the IgG by ammonium sulfate precipitation. We transfected Chinese hamster ovary cells overexpressing the AT(1) receptor with TF promoter constructs coupled to a luciferase reporter gene. VSMC were obtained from human coronary arteries. Extracellular signal-related kinase activation was detected by an in gel kinase assay. AP-1 activation was determined by electromobility shift assay. TF was measured by ELISA and detected by immunohistochemistry. Placentas from preeclamptic women stained strongly for TF, whereas control placentas showed far less staining. We proved AT(1)-AA specificity by coimmunoprecipitating the AT(1) receptor with AT(1)-AA but not with nonspecific IgG. Angiotensin (Ang) II and AT(1)-AA both activated extracellular signal-related kinase, AP-1, and the TF promoter transfected VSMC and Chinese hamster ovary cells, but only when the AP-1 binding site was present. We then demonstrated TF expression in VSMC exposed to either Ang II or AT(1)-AA. All these effects were blocked by losartan. Nonspecific IgG or IgG from nonpreeclamptic pregnant women had a negligible effect. CONCLUSIONS: We conclude that AT(1)-AA and Ang II both stimulate the AT(1) receptor and initiate a signaling cascade resulting in TF expression. These results show an action of AT(1)-AA on human cells that could contribute to the pathogenesis of preeclampsia. PMID- 10821816 TI - A common mutation in lipoprotein lipase confers a 2-fold increase in risk of ischemic cerebrovascular disease in women but not in men. AB - BACKGROUND: We previously showed that the common Asn291Ser substitution in lipoprotein lipase is associated with elevated plasma triglyceride levels and a 2 fold increase in the risk of ischemic heart disease in women but not men. In this study, we tested the hypothesis that this substitution is also associated with an increased risk of ischemic cerebrovascular disease (ICVD). METHODS AND RESULTS: We compared 260 patients who had nonfatal ICVD and carotid stenosis >/=50% with 1560 age-matched controls and also compared 205 Copenhagen City Heart Study cases who had nonfatal ICVD with 1210 age-matched controls. All subjects were white and from Denmark. Overall, no significant difference was observed between carrier frequencies among those with and without ICVD; however, sex interacted with genotype in predicting ICVD in the ICVD and carotid stenosis cases (P=0.02). In Copenhagen City Heart Study cases, sex was not significant (P=0.18). Odds ratios for ICVD in female mutation carriers were 2.9 (95% confidence interval [CI], 1.3 to 6. 4) and 1.9 (95% CI, 0.8 to 4.6) in ICVD plus carotid stenosis cases and Copenhagen City Heart Study cases, respectively. Equivalent values in male mutation carriers were 0.8 (95% CI, 0.3 to 1.8) and 0.8 (95% CI, 0.3 to 2.0), respectively. These results were similar in analyses that also allowed for other conventional cardiovascular risk factors. CONCLUSIONS: These results suggest that the Asn291Ser substitution in lipoprotein lipase is not associated with nonfatal ICVD in men but that it possibly confers a 2-fold risk in women. PMID- 10821817 TI - Heart rate dynamics at the onset of ventricular tachyarrhythmias as retrieved from implantable cardioverter-defibrillators in patients with coronary artery disease. AB - BACKGROUND: The recent availability of implantable cardioverter-defibrillators (ICDs) that record 1024 R-R intervals preceding a ventricular tachyarrhythmia (VTA) provides a unique opportunity to analyze heart rate variability (HRV) before the onset of VTA. METHODS AND RESULTS: Fifty-eight post-myocardial infarction patients with an implanted ICD for recurrent VTA provided 2 sets of 98 heart rate recordings in sinus rhythm: (1) before a VTA and (2) during control conditions. Three subgroups were considered according to the antiarrhythmic (AA) drug regimen. A state of sympathoexcitation was suggested by the significant reduction in HRV before VTA onset compared with control conditions. beta-Blockers and dl-sotalol enhanced HRV in control recordings; nevertheless, HRV declined before VTA independent of AA drugs. A gradual increase in heart rate and decrease in sinus arrhythmia at VTA onset were specific findings of patients who received dl-sotalol. CONCLUSIONS: The peculiar heart rate dynamics observed before VTA onset are suggestive of a state of sympathoexcitation that is independent of AA drugs. PMID- 10821818 TI - Significant frequencies of T cells with indirect anti-donor specificity in heart graft recipients with chronic rejection. AB - BACKGROUND: The purpose of this study was to determine whether T cells with indirect allospecificity could be detected in heart transplant recipients with chronic rejection. METHOD AND RESULTS: Human T-cell clones were used to determine the most effective way to deliver major histocompatibility complex alloantigens for indirect presentation. Seven allograft recipients with evidence of progressive, chronic rejection were selected. Four heart graft recipients with no evidence of chronic rejection were used as controls. Peripheral blood T cells and antigen-presenting cells from the recipients were cultured with frozen/thawed stored donor cells or major histocompatibility complex class I-derived synthetic peptides in limiting dilution cultures and then compared with controls using tetanus toxoid and frozen/thawed third-party cells with no human leukocyte antigens in common with the donor. In 5 of 7 patients analyzed who had chronic rejection, elevated frequencies of T cells with indirect, anti-donor specificity (iHTLf) were detected. No such elevated iHTLf were detected in recipients without chronic rejection. DISCUSSION: iHTLf can be obtained from human transplant recipients, which supports the contention that the indirect pathway is involved in chronic transplant rejection. PMID- 10821819 TI - CLA-1/SR-BI is expressed in atherosclerotic lesion macrophages and regulated by activators of peroxisome proliferator-activated receptors. AB - BACKGROUND: The scavenger receptors are cell-surface receptors for native and modified lipoproteins that play a critical role in the accumulation of lipids by macrophages. CLA-1/SR-BI binds HDL with high affinity and is involved in the cholesterol reverse-transport pathway. Peroxisome proliferator-activated receptors (PPARs) are transcription factors regulating the expression of genes implicated in lipid metabolism, cellular differentiation, and inflammation. Here, we investigated the expression of CLA-1/SR-BI in macrophages and its regulation by PPARs. METHODS AND RESULTS: CLA-1 is undetectable in human monocytes and is induced upon differentiation into macrophages. Immunohistological analysis on human atherosclerotic lesions showed high expression of CLA-1 in macrophages of the lipid core colocalizing with PPARalpha and PPARgamma staining. Activation of PPARalpha and PPARgamma resulted in the induction of CLA-1 protein expression in monocytes and in differentiated macrophages. Finally, SR-BI expression is increased in atherosclerotic lesions of apoE-null mice treated with either PPARgamma or PPARalpha ligands. CONCLUSIONS: Our data demonstrate that CLA-1/SR BI is expressed in atherosclerotic lesion macrophages and induced by PPAR activation, identifying a potential role for PPARs in cholesterol homeostasis in atherosclerotic lesion macrophages. PMID- 10821820 TI - Selective pharmacological agents implicate mitochondrial but not sarcolemmal K(ATP) channels in ischemic cardioprotection. AB - BACKGROUND: Pharmacological evidence has implicated ATP-sensitive K(+) (K(ATP)) channels as the effectors of cardioprotection, but the relative roles of mitochondrial (mitoK(ATP)) and sarcolemmal (surfaceK(ATP)) channels remain controversial. METHODS AND RESULTS: We examined the effects of the K(ATP) channel blocker HMR1098 and the K(ATP) channel opener P-1075 on surfaceK(ATP) and mitoK(ATP) channels in rabbit ventricular myocytes. HMR1098 (30 micromol/L) inhibited the surfaceK(ATP) current activated by metabolic inhibition, whereas the drug did not blunt diazoxide (100 micromol/L)-induced flavoprotein oxidation, an index of mitoK(ATP) channel activity. P-1075 (30 micromol/L) did not increase flavoprotein oxidation but did elicit a robust surfaceK(ATP) current that was completely inhibited by HMR1098. These results indicate that HMR1098 selectively inhibits surfaceK(ATP) channels, whereas P-1075 selectively activates surface K(ATP) channels. In a cellular model of simulated ischemia, the mitoK(ATP) channel opener diazoxide (100 micromol/L), but not P-1075, blunted cellular injury. The cardioprotection afforded by diazoxide or by preconditioning was prevented by the mitoK(ATP) channel blocker 5-hydroxydecanoate (500 micromol/L) but not by the surfaceK(ATP) channel blocker HMR1098 (30 micromol/L). CONCLUSIONS: The cellular effects of mitochondria- or surface-selective agents provide further support for the emerging consensus that mitoK(ATP) channels rather than surfaceK(ATP) channels are the likely effectors of cardioprotection. PMID- 10821821 TI - 99mTc-N-NOET myocardial uptake reflects myocardial blood flow and not viability in dogs with reperfused acute myocardial infarction. AB - BACKGROUND: N-Ethoxy-N-ethyl-dithiocarbamato-nitrido-(99m)Tc ((99m)Tc-N-NOET) is a new neutral lipophilic (99m)Tc-labeled myocardial perfusion agent with a high first-pass extraction fraction and delayed redistribution kinetics after transient ischemia comparable to what is observed with (201)Tl. It is unknown whether the uptake of this tracer reflects myocardial viability or just reperfusion flow in the setting of a reperfused myocardial infarction. METHODS AND RESULTS: In 13 anesthetized open-chest dogs, the left anterior descending coronary artery was occluded for 180 minutes, followed by 180 minutes of reperfusion. (201)Tl and (99m)Tc-N-NOET were injected after either 60 (group 1, n=9) or 175 (group 2, n=4) minutes of reperfusion. Myocardial blood flow was measured by radioactive microspheres, and (201)Tl and (99m)Tc-N-NOET tissue activities were determined by gamma-well counting. Normalized myocardial blood flow in the central infarct zone fell from 0.80+/-0. 03 (SEM) and 0.89+/-0.01 at baseline to 0.18+/-0.04 and 0.13+/-0.02 during the occlusion in groups 1 and 2, respectively. Normalized (201)Tl activity in these segments was 0.39+/-0.04 and 0.43+/-0.04 and reflected myocardial viability rather than reperfusion flow (P<0. 001). Normalized (99m)Tc-N-NOET activity in the same segments was 0. 84+/-0.08 and 0.64+/-0.03, respectively (P<0.01 versus (201)Tl; P=NS versus reperfusion flow) and more accurately reflected reperfusion flow (0.99+/-0.17 and 0.70+/ 0.04) than residual viability. CONCLUSIONS: The myocardial uptake of (99m)Tc-N NOET reflects reperfusion myocardial blood flow and not viability in a canine model of reperfused acute myocardial infarction. The clinical use of early (99m)Tc-N-NOET imaging to assess the success of coronary reperfusion in patients with acute myocardial infarction should be investigated. PMID- 10821823 TI - Mechanism of ventricular defibrillation. The role of tissue geometry in the changes in transmembrane potential in patterned myocyte cultures. AB - BACKGROUND: The geometry of the myocardium may influence changes in transmembrane potential (DeltaVm) during defibrillation. To test this hypothesis, specific nonlinear structures (bifurcations, expansions, and curved strands or "bends") were created in patterned cultures of neonatal rat myocytes. METHODS AND RESULTS: Extracellular field stimuli (EFS; 7 to 11 V/cm field strength) were applied parallel to the strands. Changes in Vm were measured with microscopic resolution using optical mapping techniques. In bifurcations, EFS produced 2 DeltaVm maxima (so-called secondary sources) at the shoulder of each limb that were separated by a decrease of either hyperpolarization or depolarization at the insertion of the stem strand. In expansions, EFS produced a significant decrease in DeltaVm at the insertion site of the expansion compared with the DeltaVm maxima measured at the lateral borders. In 50% of experiments, tertiary sources of opposite polarity appeared in the strand due to local electrotonic currents. New action potentials were propagated from the sites of DeltaVm maxima located at the lateral borders of the expansions. In bends, the strand oriented in parallel to the field dominated electrotonically and partially cancelled the sources produced by the perpendicular segment. CONCLUSIONS: In electrically well-coupled nonlinear structures, EFS produced changes in Vm at resistive boundaries that were determined by the electrotonic interaction between sources of different, direction-dependent strength. In addition, the interaction between localized secondary sources at nonlinear boundaries generated local current circuits, which gave rise to further changes in Vm (tertiary sources). PMID- 10821822 TI - Calcineurin expression, activation, and function in cardiac pressure-overload hypertrophy. AB - BACKGROUND: Vascular hypertension resulting in increased cardiac load is associated with left ventricular hypertrophy and is a leading predicator for progressive heart disease. The molecular signaling pathways that respond to increases in cardiac load are poorly understood. One potential regulator of the hypertrophic response is the calcium-sensitive phosphatase calcineurin. METHODS AND RESULTS: We showed that calcineurin enzymatic activity is increased 3. 2-fold in the heart in response to pressure-overload hypertrophy induced by abdominal aortic banding in the rat. Western blot analysis further demonstrates that calcineurin A (catalytic subunit) protein content and association with calmodulin are increased in response to pressure-overload hypertrophy. This increase in calcineurin protein content was prevented by administration of the calcineurin inhibitor cyclosporine A (CsA). CsA administration attenuated load-induced cardiac hypertrophy in a dose-dependent manner over a 14-day treatment protocol. CsA administration also partially reversed pressure-overload hypertrophy in aortic-banded rats after 14 days. CsA also attenuated the histological and molecular indexes of pressure-overload hypertrophy. CONCLUSIONS: These data suggest that calcineurin is an important upstream regulator of load-induced hypertrophy. PMID- 10821825 TI - Oculostenotic reflex and iatrogenosis fulminans. PMID- 10821824 TI - Tangier disease with continuous massive and longitudinal diffuse calcification in the coronary arteries : demonstration by the sagittal images of intravascular ultrasonography. PMID- 10821826 TI - Global tuberculosis threat demands US response. PMID- 10821827 TI - New insights into the size and stoichiometry of the plasminogen activator inhibitor type-1.vitronectin complex. AB - Plasminogen activator inhibitor-type 1 (PAI-1) is the primary inhibitor of endogenous plasminogen activators that generate plasmin in the vicinity of a thrombus to initiate thrombolysis, or in the pericellular region of cells to facilitate migration and/or tissue remodeling. It has been shown that the physiologically relevant form of PAI-1 is in a complex with the abundant plasma glycoprotein, vitronectin. The interaction between vitronectin and PAI-1 is important for stabilizing the inhibitor in a reactive conformation. Although the complex is clearly significant, information is vague regarding the composition of the complex and consequences of its formation on the distribution and activity of vitronectin in vivo. Most studies have assumed a 1:1 interaction between the two proteins, but this has not been demonstrated experimentally and is a matter of some controversy since more than one PAI-1-binding site has been proposed within the sequence of vitronectin. To address this issue, competition studies using monoclonal antibodies specific for separate epitopes confirmed that the two distinct PAI-1-binding sites present on vitronectin can be occupied simultaneously. Analytical ultracentrifugation was used also for a rigorous analysis of the composition and sizes of complexes formed from purified vitronectin and PAI-1. The predominant associating species observed was high in molecular weight (M(r) approximately 320,000), demonstrating that self association of vitronectin occurs upon interaction with PAI-1. Moreover, the size of this higher order complex indicates that two molecules of PAI-1 bind per vitronectin molecule. Binding of PAI-1 to vitronectin and association into higher order complexes is proposed to facilitate interaction with macromolecules on surfaces. PMID- 10821828 TI - Elevation of mitochondrial calcium by ryanodine-sensitive calcium-induced calcium release. AB - Calcium is an important regulator of mitochondrial function. Since there can be tight coupling between inositol 1,4, 5-trisphosphate-sensitive Ca(2+) release and elevation of mitochondrial calcium concentration, we have investigated whether a similar relationship exists between the release of Ca(2+) from the ryanodine receptor and the elevation of mitochondrial Ca(2+). Perfusion of permeabilized A10 cells with inositol 1,4, 5-trisphosphate resulted in a large transient elevation of mitochondrial Ca(2+) to about 8 microm. The response was inhibited by heparin but not ryanodine. Perfusion of the cells with Ca(2+) buffers in excess of 1 microm leads to large increases in mitochondrial Ca(2+) that are much greater than the perfused Ca(2+). These increases, which average around 10 microm, are enhanced by caffeine and inhibited by ryanodine and depletion of the intracellular stores with either orthovanadate or thapsigargin. We conclude that Ca(2+)-induced Ca(2+) release at the ryanodine receptor generates microdomains of elevated Ca(2+) that are sensed by adjacent mitochondria. In addition to ryanodine-sensitive stores acting as a source of Ca(2+), Ca(2+)-induced Ca(2+) release is required to generate efficient elevation of mitochondrial Ca(2+). PMID- 10821829 TI - Inhibition of selenoprotein synthesis by selenocysteine tRNA[Ser]Sec lacking isopentenyladenosine. AB - A common posttranscriptional modification of tRNA is the isopentenylation of adenosine at position 37, creating isopentenyladenosine (i(6)A). The role of this modified nucleoside in protein synthesis of higher eukaryotes is not well understood. Selenocysteyl (Sec) tRNA (tRNA([Ser]Sec)) decodes specific UGA codons and contains i(6)A. To address the role of the modified nucleoside in this tRNA, we constructed a site-specific mutation, which eliminates the site of isopentenylation, in the Xenopus tRNA([Ser]Sec) gene. Transfection of the mutant tRNA([Ser]Sec) gene resulted in 80% and 95% reduction in the expression of co transfected selenoprotein genes encoding type I and II iodothyronine deiodinases, respectively. A similar decrease in type I deiodinase synthesis was observed when transfected cells were treated with lovastatin, an inhibitor of the biosynthesis of the isopentenyl moiety. Neither co-transfection with the mutant tRNA gene nor lovastatin treatment reduced type I deiodinase mRNA levels. Also, mutant tRNA expression did not alter initiation of translation or degradation of the type I deiodinase protein. Furthermore, isopentenylation of tRNA([Ser]Sec) was not required for synthesis of Sec on the tRNA. We conclude that isopentenylation of tRNA([Ser]Sec) is required for efficient translational decoding of UGA and synthesis of selenoproteins. PMID- 10821830 TI - Self-assembly and supramolecular organization of EMILIN. AB - The primary structure of human Elastin microfibril interface-located protein (EMILIN), an elastic fiber-associated glycoprotein, consists of a globular C1q domain (gC1q) at the C terminus, a short collagenous stalk, a long region with a high potential for forming coiled-coil alpha helices, and a cysteine-rich N terminal sequence. It is not known whether the EMILIN gC1q domain is involved in the assembly process and in the supramolecular organization as shown for the similar domain of collagen X. By employing the yeast two-hybrid system the EMILIN gC1q domains interacted with themselves, proving for the first time that this interaction occurs in vivo. The gC1q domain formed oligomers running as trimers in native gels that were less stable than the comparable trimers of the collagen X gC1q domain since they did not withstand heating. The collagenous domain was trypsin-resistant and migrated at a size corresponding to a triple helix under native conditions. In reducing agarose gels, EMILIN also migrated as a trimer, whereas under non-reducing conditions it formed polymers of many millions of daltons. A truncated fragment lacking gC1q and collagenous domains assembled to a much lesser extent, thus deducing that the C-terminal domain(s) are essential for the formation of trimers that finally assemble into large EMILIN multimers. PMID- 10821831 TI - Partial deletion of a loop region in the high affinity K+ transporter HKT1 changes ionic permeability leading to increased salt tolerance. AB - HKT1 is a high affinity K(+) transporter protein that is a member of a large superfamily of transporters found in plants, bacteria, and fungi. These transporters are primarily involved in K(+) uptake and are energized by Na(+) or H(+). HKT1 is energized by Na(+) but also mediates low affinity Na(+) uptake and may therefore be a pathway for Na(+) uptake, which is toxic to plants. The aim of this study was to identify regions of HKT1 that are involved in K(+)/Na(+) selectivity and alter the amino acid composition in those regions to increase the ionic selectivity of the transporter. A highly charged loop was identified, and two deletions were created that resulted in the removal of charged and uncharged amino acids. The functional changes caused by the deletions were studied in yeast and Xenopus oocytes. The deletions improved the K(+)/Na(+) selectivity of the transporter and increased the salt tolerance of the yeast cells in which they were expressed. In light of recent structural models of members of this symporter superfamily, it was necessary to determine the orientation of this highly charged loop. Introduction of an epitope tag allowed us to demonstrate that this loop faces the outside of the membrane where it is likely to facilitate the interaction with cations such as K(+) and Na(+). This study has identified an important structural feature in HKT1 that in part determines its K(+)/Na(+) selectivity. Understanding the structural basis of the functional characteristics in transporters such as HKT1 may have important implications for increasing the salt tolerance of higher plants. PMID- 10821832 TI - Topological analysis of Niemann-Pick C1 protein reveals that the membrane orientation of the putative sterol-sensing domain is identical to those of 3 hydroxy-3-methylglutaryl-CoA reductase and sterol regulatory element binding protein cleavage-activating protein. AB - The Niemann-Pick C1 (NPC1) protein is predicted to be a polytopic glycoprotein, and it contains a region with extensive homology to the sterol-sensing domains (SSD) of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-R) and sterol regulatory element binding protein cleavage-activating protein (SCAP). To aid the functional characterization of NPC1, a model of NPC1 topology was evaluated by expression of epitope-tagged NPC1 proteins and investigation of epitope accessibility in selectively permeabilized cells. These results were further confirmed by expression of NPC1 and identification of glycosylated domains that are located in the lumen of the endoplasmic reticulum. Our data indicate that this glycoprotein contains 13 transmembrane domains, 3 large and 4 small luminal loops, 6 small cytoplasmic loops, and a cytoplasmic tail. Furthermore, our data show that the putative SSD of NPC1 is oriented in the same manner as those of HMG R and SCAP, providing strong evidence that this domain is functionally important. PMID- 10821833 TI - Nucleotide excision repair of the 5 S ribosomal RNA gene assembled into a nucleosome. AB - A-175-base pair fragment containing the Xenopus borealis somatic 5 S ribosomal RNA gene was used as a model system to determine the effect of nucleosome assembly on nucleotide excision repair (NER) of the major UV photoproduct (cyclobutane pyrimidine dimer (CPD)) in DNA. Xenopus oocyte nuclear extracts were used to carry out repair in vitro on reconstituted, positioned 5 S rDNA nucleosomes. Nucleosome structure strongly inhibits NER at many CPD sites in the 5 S rDNA fragment while having little effect at a few sites. The time course of CPD removal at 35 different sites indicates that >85% of the CPDs in the naked DNA fragment have t(12) values <2 h, whereas <26% of the t(12) values in nucleosomes are <2 h, and 15% are >8 h. Moreover, removal of histone tails from these mononucleosomes has little effect on the repair rates. Finally, nucleosome inhibition of repair shows no correlation with the rotational setting of a 14 nucleotide-long pyrimidine tract located 30 base pairs from the nucleosome dyad. These results suggest that inhibition of NER by mononucleosomes is not significantly influenced by the rotational orientation of CPDs on the histone surface, and histone tails play little (or no) role in this inhibition. PMID- 10821834 TI - The cytosolic termini of the beta- and gamma-ENaC subunits are involved in the functional interactions between cystic fibrosis transmembrane conductance regulator and epithelial sodium channel. AB - Epithelial sodium channel (ENaC) and cystic fibrosis transmembrane conductance regulator (CFTR) are co-localized in the apical membrane of many epithelia. These channels are essential for electrolyte and water secretion and/or reabsorption. In cystic fibrosis airway epithelia, a hyperactivated epithelial Na(+) conductance operates in parallel with defective Cl(-) secretion. Several groups have shown that CFTR down-regulates ENaC activity, but the mechanisms and the regulation of CFTR by ENaC are unknown. To test the hypothesis that ENaC and CFTR regulate each other, and to identify the region(s) of ENaC involved in the interaction between CFTR and ENaC, rENaC and its mutants were co-expressed with CFTR in Xenopus oocytes. Whole cell macroscopic sodium currents revealed that wild type (wt) alphabetagamma-rENaC-induced Na(+) current was inhibited by co expression of CFTR, and further inhibited when CFTR was activated with a cAMP raising mixture (CKT). Conversely, alphabetagamma-rENaC stimulated CFTR-mediated Cl(-) currents up to approximately 6-fold. Deletion mutations in the intracellular tails of the three rENaC subunits suggested that the carboxyl terminus of the beta subunit was required both for the down-regulation of ENaC by activated CFTR and the up-regulation of CFTR by ENaC. However, both the carboxyl terminus of the beta subunit and the amino terminus of the gamma subunit were essential for the down-regulation of rENaC by unstimulated CFTR. Interestingly, down-regulation of rENaC by activated CFTR was Cl(-)-dependent, while stimulation of CFTR by rENaC was not dependent on either cytoplasmic Na(+) or a depolarized membrane potential. In summary, there appear to be at least two different sites in ENaC involved in the intermolecular interaction between CFTR and ENaC. PMID- 10821835 TI - Adhesion of human skin fibroblasts to Cyr61 is mediated through integrin alpha 6beta 1 and cell surface heparan sulfate proteoglycans. AB - The angiogenic inducer Cyr61 is an extracellular matrix-associated heparin binding protein that can mediate cell adhesion, stimulate cell migration, and enhance growth factor-stimulated DNA synthesis in both fibroblasts and endothelial cells in culture. In vivo, Cyr61 induces neovascularization and promotes tumor growth. Cyr61 is a prototypic member of a highly conserved family of secreted proteins that includes connective tissue growth factor, nephroblastoma overexpressed, Elm-1/WISP-1, Cop-1/WISP-2, and WISP-3. Encoded by an immediate early gene, Cyr61 synthesis is induced by serum growth factors in cultured fibroblasts and in dermal fibroblasts during cutaneous wound healing. We previously demonstrated that Cyr61 mediates adhesion of vascular endothelial cells and activation-dependent adhesion of blood platelets through direct interaction with integrins alpha(V)beta(3) and alpha(IIb)beta(3), respectively. In this study, we show that the adhesion of primary human skin fibroblasts to Cyr61 is mediated through integrin alpha(6)beta(1) and cell surface heparan sulfate proteoglycans (HSPGs), which most likely serve as co-receptors. Either destruction of cell surface HSPGs or prior occupancy of the Cyr61 heparin-binding site completely blocked cell adhesion to Cyr61. A heparin-binding defective mutant of Cyr61 was unable to mediate fibroblast adhesion through integrin alpha(6)beta(1) but still mediated endothelial cell adhesion through integrin alpha(V)beta(3), indicating that endothelial cell adhesion through integrin alpha(V)beta(3) is independent of the heparin-binding activity of Cyr61. These results identify Cyr61 as a novel adhesive substrate for integrin alpha(6)beta(1) and provide the first demonstration of the requirement for HSPGs in integrin mediated cell attachment. In addition, these findings suggest that Cyr61 might elicit disparate biological effects in different cell types through interaction with distinct integrin receptors. PMID- 10821836 TI - Multiple S gene family members including natural antisense transcripts are differentially expressed during development of maize flowers. AB - Within the large Brassica S gene family, SLG (S locus glycoprotein) and SRK (S locus receptor kinase) participate to the control of pollen-stigma self incompatibility. In the self-compatible species maize, S gene family members are predominantly expressed in vegetative organs but are also expressed to a lesser extent in the stigma (silk). To determine if the expression of any S gene family members correlates with female receptivity, we analyzed their expression in developing maize silks. We show that a large family of maize S transcripts is expressed in developing silks. Surprisingly, we isolated a cDNA complementary to a large portion of the antisense strand of the maize receptor kinase S domain. Rapid amplification of cDNA ends (RACE)-polymerase chain reaction, RNase protection, and Northern hybridization with single-stranded riboprobes confirmed that natural antisense S transcripts exist in leaves and seedling shoots and in all sexual tissues tested except mature pollen. These natural antisense S transcripts co-exist with several less abundant sense S transcripts. The accumulation of sense and antisense S transcripts is differentially regulated during pollen and silk development. Thus, these results support a role for S gene family members in sexual tissue development and/or compatible pollination and reveal a new level of complexity in the regulation and function of the S gene family in maize. PMID- 10821837 TI - Kinetics and the mechanism of interaction of the endoplasmic reticulum chaperone, calreticulin, with monoglucosylated (Glc1Man9GlcNAc2) substrate. AB - Calreticulin is a lectin-like molecular chaperone of the endoplasmic reticulum in eukaryotes. Its interaction with N-glycosylated polypeptides is mediated by the glycan, Glc(1)Man(9)GlcNAc(2), present on the target glycoproteins. In this work, binding of monoglucosyl IgG (chicken) substrate to calreticulin has been studied using real time association kinetics of the interaction with the biosensor based on surface plasmon resonance (SPR). By SPR, accurate association and dissociation rate constants were determined, and these yielded a micromolar association constant. The nature of reaction was unaffected by immobilization of either of the reactants. The Scatchard analysis values for K(a) agreed well with the one obtained by the ratio k(1)/k(-1). The interaction was completely inhibited by free oligosaccharide, Glc(1)Man(9)GlcNAc(2,) whereas Man(9)GlcNAc(2) did not bind to the calreticulin-substrate complex, attesting to the exquisite specificity of this interaction. The binding of calreticulin to IgG was used for the development of immunoassay and the relative affinity of the lectin-substrate association was indirectly measured. The values are in agreement with those obtained with SPR. Although the reactions are several orders of magnitude slower than the diffusion controlled processes, the data are qualitatively and quantitatively consistent with single-step bimolecular association and dissociation reaction. Analyses of the activation parameters indicate that reaction is enthalpically driven and does not involve a highly ordered transition state. Based on these data, the mechanism of its chaperone activity is briefly discussed. PMID- 10821838 TI - Substitutions at codon 22 of Alzheimer's abeta peptide induce diverse conformational changes and apoptotic effects in human cerebral endothelial cells. AB - Cerebral amyloid angiopathy is commonly associated with normal aging and Alzheimer's disease and it is also the principal feature of hereditary cerebral hemorrhage with amyloidosis Dutch type, a familial condition associated to a point mutation G to C at codon 693 of the amyloid beta (Abeta) precursor protein gene resulting in a Glu to Gln substitution at position 22 of the Abeta (E22Q). The patients carrying the AbetaE22Q variant usually present with lobar cerebral hemorrhages before 50 years of age. A different mutation described in several members of three Italian kindred who presented with recurrent hemorrhagic strokes late in life, between 60 and 70 years of age, also associated with extensive cerebrovascular amyloid deposition has been found at the same position 22, this time resulting in a Glu to Lys substitution (E22K). We have compared the secondary structure, aggregation, and fibrillization properties of the two Abeta40 variants and the wild type peptide. Using flow cytometry analysis after staining with propidium iodide and annexin V, we also evaluated the cytotoxic effects of the peptides on human cerebral endothelial cells in culture. Under the conditions tested, the E22Q peptide exhibited the highest content of beta-sheet conformation and the fastest aggregation/fibrillization properties. The Dutch variant also induced apoptosis of cerebral endothelial cells at a concentration of 25 micrometer, whereas the wild type Abeta and the E22K mutant had no effect. The data suggest that different amino acids at position 22 confer distinct structural properties to the peptides that appear to influence the onset and aggressiveness of the disease rather than the phenotype. PMID- 10821839 TI - Development and characterization of a recombinant truncated type VII collagen "minigene". Implication for gene therapy of dystrophic epidermolysis bullosa. AB - Dystrophic epidermolysis bullosa (DEB) is an inherited mechano-bullous disorder of skin caused by mutations in the type VII collagen gene. The lack of therapy for DEB provides an impetus to develop gene therapy strategies. However, the full length 9-kilobase type VII collagen cDNA exceeds the cloning capacity of current viral delivery vectors. In this study, we produced a recombinant type VII minicollagen containing the intact noncollagenous domains, NC1 and NC2, and part of the central collagenous domain using stably transfected human 293 cell clones and purified large quantities of the recombinant minicollagen VII from culture media. Minicollagen VII was secreted as correctly-folded, disulfide-bonded, helical trimers resistant to protease degradation. Purified minicollagen VII bound to fibronectin, laminin-5, type I collagen, and type IV collagen. Furthermore, retroviral-mediated transduction of the minigene construct into DEB keratinocytes (in which type VII collagen was absent) resulted in persistent synthesis and secretion of a 230-kDa recombinant minicollagen VII. In comparison with parent DEB keratinocytes, the gene-corrected DEB keratinocytes demonstrated enhanced cell-substratum adhesion, increased proliferative potential, and reduced cell motility, features that reversed the DEB phenotype toward normal. We conclude that the use of the minicollagen VII may provide a strategy to correct the cellular manifestations of gene defects in DEB. PMID- 10821840 TI - Role of arginine 285 in the active site of Rhodotorula gracilis D-amino acid oxidase. A site-directed mutagenesis study. AB - Arg(285), one of the very few conserved residues in the active site of d-amino acid oxidases, has been mutated to lysine, glutamine, aspartate, and alanine in the enzyme from the yeast Rhodotorula gracilis (RgDAAO). The mutated proteins are all catalytically competent. Mutations of Arg(285) result in an increase ( approximately 300-fold) of K(m) for the d-amino acid and in a large decrease ( approximately 500-fold) of turnover number. Stopped-flow analysis shows that the decrease in turnover is paralleled by a similar decrease in the rate of flavin reduction (k(2)), the latter still being the rate-limiting step of the reaction. In agreement with data from the protein crystal structure, loss of the guanidinium group of Arg(285) in the mutated DAAOs drastically reduces the binding of several carboxylic acids (e.g. benzoate). These results highlight the importance of this active site residue in the precise substrate orientation, a main factor in this redox reaction. Furthermore, Arg(285) DAAO mutants have spectral properties similar to those of the wild-type enzyme, but show a low degree of stabilization of the flavin semiquinone and a change in the redox properties of the free enzyme. From this, we can unexpectedly conclude that Arg(285) in the free enzyme form is involved in the stabilization of the negative charge on the N(1)-C(2)=O locus of the isoalloxazine ring of the flavin. We also suggest that the residue undergoes a conformational change in order to bind the carboxylate portion of the substrate/ligand in the complexed enzyme. PMID- 10821841 TI - G13alpha-mediated PYK2 activation. PYK2 is a mediator of G13alpha -induced serum response element-dependent transcription. AB - G(12)alpha/G(13)alpha transduces signals from G-protein-coupled receptors to stimulate growth-promoting pathways and the early response gene c-fos. Within the c-fos promoter lies a key regulatory site, the serum response element (SRE). Here we show a critical role for the tyrosine kinase PYK2 in muscarinic receptor type 1 and G(12)alpha/G(13)alpha signaling to an SRE reporter gene. A kinase inactivate form of PYK2 (PYK2 KD) inhibits muscarinic receptor type 1 signaling to the SRE and PYK2 itself triggers SRE reporter gene activation through a RhoA dependent pathway. Placing PYK2 downstream of G-protein activation but upstream of RhoA, the expression of PYK2 KD blocks the activation of an SRE reporter gene by GTPase-deficient forms of G(12)alpha or G(13)alpha but not by RhoA. The GTPase deficient form of G(13)alpha triggers PYK2 kinase activity and PYK2 tyrosine phosphorylation, and co-expression of the RGS domain of p115 RhoGEF inhibits both responses. Finally, we show that in vivo G(13)alpha, although not G(12)alpha, readily associates with PYK2. Thus, G-protein-coupled receptors via G(13)alpha activation can use PYK2 to link to SRE-dependent gene expression. PMID- 10821842 TI - The promoter for constitutive expression of the human ICln gene CLNS1A. AB - The ICln protein is expressed ubiquitously in mammals. Experiments designed to knock down the ICln protein in NIH 3T3 fibroblasts as well as in epithelial cells led to the conclusion that this protein is crucially involved in volume regulation after cytoplasmic swelling. Reconstitution of the ICln protein in lipid bilayers revealed the ion channel nature of ICln. Here we describe a new human promoter sequence, composed of 89 nucleotides, which is responsible for a highly constitutive expression of the ICln protein. The promoter sequence lacks a TATA box, and the transcription can be effected at multiple sites. In addition to the starting sites, upstream sequence elements are mandatory for an efficient transcription of the ICln gene (CLNS1A). These new nucleotide elements were defined by site-directed mutagenesis. PMID- 10821843 TI - Genomic and functional changes induced by the activation of the peripheral cannabinoid receptor CB2 in the promyelocytic cells HL-60. Possible involvement of the CB2 receptor in cell differentiation. AB - The function of the peripheral cannabinoid receptor (CB2), which is mainly expressed on hematopoietic cells, remains an enigma. In an attempt to decipher its role, we used Affymetrix DNA chips to investigate the gene expression profile of the promyelocytic cells HL-60 transfected with the CB2 receptor and activated with the cannabinoid agonist CP 55,940. Agonist exposure of these cells led to an activation of a mitogen-activated protein kinase cascade and a receptor desensitization, indicating a functional coupling of the transfected receptors. At the genomic level, activation of the CB2 receptors induced an up-regulation of nine genes involved in cytokine synthesis, regulation of transcription, and cell differentiation. A majority of them are under the control of the transcription factor NF-kappaB, whose nuclear translocation was demonstrated. Many features of the transcriptional events, reported here for the first time, appeared to be related to an activation of a cell differentiation program, suggesting that CB2 receptors could play a role in the initialization of cell maturation. Moreover, we showed that CB2-activated wild-type HL-60 cells developed properties usually found in host defense effector cells such as an enhanced release of chemotactic cytokines and an increased motility, characteristic of more mature cells of the granulocytic-monocytic lineage. PMID- 10821844 TI - Alpha-melanocyte-stimulating hormone reduces impact of proinflammatory cytokine and peroxide-generated oxidative stress on keratinocyte and melanoma cell lines. AB - We have previously shown that alpha-melanocyte-stimulating hormone (alpha-MSH) can oppose tumor necrosis factor alpha activation of NF-kappaB (1-2 h) and intercellular adhesion molecule 1 up-regulation (mRNA by 3 h and protein by 24 h) in melanocytes and melanoma cells. The present study reports on the ability of four MSH peptides to control intracellular peroxide levels and glutathione peroxidase (GPx) activity in pigmentary and nonpigmentary cells. In human HBL melanoma and HaCaT keratinocytes tumor necrosis factor alpha and H(2)O(2) both activated GPx in a time- and concentration-dependent manner (by 30-45 min). alpha MSH peptides were found to inhibit the stimulated GPx activity and had biphasic dose-response curves. MSH 1-13 and MSH [Nle(4)-d-Phe(7)] achieved maximum inhibition at 10(-10) and 10(-12) m, respectively. Higher concentrations (10-100 fold) of MSH 4-10 and MSH 11-13 were required to produce equivalent levels of inhibition. alpha-MSH was also capable of reducing peroxide accumulation within 15 min, and again this inhibition was biphasic. The data support a role of alpha MSH in acute protection of cells to oxidative/cytokine action that precedes NF kappaB and GPx activation. The rapidity and potency of the response to alpha-MSH in pigmentary and nonpigmentary cells suggest this to be a central role of this peptide in cutaneous cells. PMID- 10821845 TI - The activating component of the anaerobic ribonucleotide reductase from Escherichia coli. An iron-sulfur center with only three cysteines. AB - Class III anaerobic ribonucleotide reductase small component, named protein beta, contains a (4Fe-4S) center. Its function is to mediate electron transfer from reduced flavodoxin to S-adenosylmethionine, required for the introduction of a glycyl radical in the large component, named protein alpha, which then becomes active for the reduction of ribonucleotides. By site-directed mutagenesis we demonstrate that the three cysteines of the conserved CXXXCXXC sequence are involved in iron chelation. Such a sequence is also present in the activase of the pyruvate formate-lyase and in the biotin synthase, both carrying an iron sulfur center involved in reductive activation of S-adenosylmethionine. Even though they are able to bind iron in the (4Fe-4S) form, as shown by Mossbauer spectroscopy, the corresponding Cys to Ala mutants are catalytically inactive. Mutation of the two other cysteines of the protein did not result in inactivation. We thus conclude that the (4Fe-4S) cluster has, in the wild type protein, only three cysteine ligands and a fourth still unidentified ligand. PMID- 10821846 TI - Nod factor requirements for efficient stem and root nodulation of the tropical legume Sesbania rostrata. AB - Azorhizobium caulinodans ORS571 synthesizes mainly pentameric Nod factors with a household fatty acid, an N-methyl, and a 6-O-carbamoyl group at the nonreducing terminal residue and with a d-arabinosyl, an l-fucosyl group, or both at the reducing-terminal residue. Nodulation on Sesbania rostrata was carried out with a set of bacterial mutants that produce well characterized Nod factor populations. Purified Nod factors were tested for their capacity to induce root hair formation and for their stability in an in vitro degradation assay with extracts of uninfected adventitious rootlets. The glycosylations increased synergistically the nodulation efficiency and the capacity to induce root hairs, and they protected the Nod factor against degradation. The d-arabinosyl group was more important than the l-fucosyl group for nodulation efficiency. Replacement of the 6-O-l-fucosyl group by a 6-O-sulfate ester did not affect Nod factor stability, but reduced nodulation efficiency, indicating that the l-fucosyl group may play a role in recognition. The 6-O-carbamoyl group contributes to nodulation efficiency, biological activity, and protection, but could be replaced by a 6-O acetyl group for root nodulation. The results demonstrate that none of the studied substitutions is strictly required for triggering normal nodule formation. However, the nodulation efficiency was greatly determined by the synergistic presence of substitutions. Within the range tested, fluctuations of Nod factor amounts had little impact on the symbiotic phenotype. PMID- 10821847 TI - Identification of a serine hydrolase as a key determinant in the microbial degradation of polychlorinated biphenyls. AB - The ability of 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoate (HOPDA) hydrolase (BphD) of Burkholderia cepacia LB400 to hydrolyze polychlorinated biphenyl (PCB) metabolites was assessed by determining its specificity for monochlorinated HOPDAs. The relative specificities of BphD for HOPDAs bearing chlorine substituents on the phenyl moiety were 0.28, 0.38, and 1.1 for 8-Cl, 9-Cl, and 10 Cl HOPDA, respectively, versus HOPDA (100 mm phosphate, pH 7.5, 25 degrees C). In contrast, HOPDAs bearing chlorine substituents on the dienoate moiety were poor substrates for BphD, which hydrolyzed 3-Cl, 4-Cl, and 5-Cl HOPDA at relative maximal rates of 2.1 x 10(-3), 1.4 x 10(-4), and 0.36, respectively, versus HOPDA. The enzymatic transformation of 3-, 5-, 8-, 9-, and 10-Cl HOPDAs yielded stoichiometric quantities of the corresponding benzoate, indicating that BphD catalyzes the hydrolysis of these HOPDAs in the same manner as unchlorinated HOPDA. HOPDAs also underwent a nonenzymatic transformation to products that included acetophenone. In the case of 4-Cl HOPDA, this transformation proceeded via the formation of 4-OH HOPDA (t(12) = 2.8 h; 100 mm phosphate, pH 7.5, 25 degrees C). 3-Cl HOPDA (t(12) = 504 h) was almost 3 times more stable than 4-OH HOPDA. Finally, 3-Cl, 4-Cl and 4-OH HOPDAs competitively inhibited the BphD catalyzed hydrolysis of HOPDA (K(ic) values of 0.57 +/- 0. 04, 3.6 +/- 0.2, and 0.95 +/- 0.04 microm, respectively). These results explain the accumulation of HOPDAs and chloroacetophenones in the microbial degradation of certain PCB congeners. More significantly, they indicate that in the degradation of PCB mixtures, BphD would be inhibited, thereby slowing the mineralization of all congeners. BphD is thus a key determinant in the aerobic microbial degradation of PCBs. PMID- 10821848 TI - Biochemical structural analysis of the lantibiotic mutacin II. AB - Mutacin II is a post-translationally modified lantibiotic peptide secreted by Streptococcus mutans T8, which inhibits the energy metabolism of sensitive cells. The deduced amino acid sequence of promutacin II is NRWWQGVVPTVSYECRMNSWQHVFTCC, which is capable of forming three thioether bridges. It was not obvious, however, how the three thioether bridges are organized. To examine the bridging, the cyanogen bromide cleavage products of mutacin II and its variants generated by protein engineering, C15A, C26A, and C15A/C26A, were analyzed by mass spectrometry. Analysis of the wild type molecule and the C15A variant excluded several possibilities and also indicated a high fidelity of formation of the thioether bridges. This allowed us to further resolve the structure by analysis (mass spectrometry and tandem mass spectrometry) of the cyanogen bromide cleavage fragments of the C26A and C15A/C26A mutants. Nuclear magnetic resonance analysis established the presence of one and two dehydrobutyrine residues in mutacin II and the C15A variant, respectively, thus yielding the final structure. The results of this investigation showed that the C-terminal part contains three thioether bridges connecting Cys residues 15, 26, and 27 to Ser/Thr residues 10, 12 and 19, respectively, with Thr(25) being modified to dehydrobutyrine. PMID- 10821849 TI - Chrysoptin is a potent glycoprotein IIb/IIIa fibrinogen receptor antagonist present in salivary gland extracts of the deerfly. AB - Salivary gland lysates of the deerfly (genus Chrysops) contain chrysoptin, an inhibitor of ADP-induced platelet aggregation, which presumably assists the fly in obtaining a blood meal. Chrysoptin has now been isolated, and its cDNA has been cloned and expressed. Chrysoptin was purified to homogeneity using anion exchange and hydrophobic interaction chromatography and found to be a protein with a molecular mass of 65 kDa as determined by gel electrophoresis. N-terminal amino acid sequencing allowed for the synthesis of degenerate oligonucleotides that led to cloning, from salivary gland specific mRNA, of the cDNA encoding this platelet inhibitor. No RGD sites are present in the predicted sequence. A search of GenBank(TM) did not reveal significant sequence homology between chrysoptin and other proteins. The molecular mass predicted from the cDNA was 59 kDa. Predicted glycosylation and phosphorylation sites may account for this difference in molecular mass, as recombinant chrysoptin expressed in Sf21 cells had a molecular mass of 65 kDa, matching that of the natural protein. Chrysoptin functions by inhibiting the binding of fibrinogen to the fibrinogen/glycoprotein IIb/IIIa receptor on platelets with an IC(50) of 95 pmol. These results reveal that insect salivary glands are a source of fibrinogen receptor antagonists. PMID- 10821850 TI - Divergent hTAFII31-binding motifs hidden in activation domains. AB - Activation domains are functional modules that enable DNA-binding proteins to stimulate transcription. Characterization of these essential modules in transcription factors has been hampered by their low sequence homology. Here we delineate the peptide sequences that are required for transactivation and interaction with hTAF(II)31, a classical target of the acidic class of activation domains. Our analyses indicate that hTAF(II)31 recognizes a diverse set of sequences for transactivation. This information enabled the identification of hTAF(II)31-binding sequences that are critical for the activity of the activation domains of five human transcription factors: NFAT1, ALL1, NF-IL6, ESX, and HSF-1. The interaction surfaces are localized in short peptide segments of activation domains. The brevity and heterogeneity of the motifs may explain the low sequence homology among acidic activation domains. PMID- 10821851 TI - Glucose-stimulated preproinsulin gene expression and nuclear trans-location of pancreatic duodenum homeobox-1 require activation of phosphatidylinositol 3 kinase but not p38 MAPK/SAPK2. AB - Exposure of islet beta-cells to elevated glucose concentrations (30 versus 3 mm) prompts enhanced preproinsulin (PPI) gene transcription and the trans-location to the nucleoplasm of pancreatic duodenum homeobox-1 (PDX-1; Rafiq, I., Kennedy, H., and Rutter, G. A. (1998) J. Biol. Chem. 273, 23241-23247). Here, we show that in MIN6 beta-cells, over-expression of p110.CAAX, a constitutively active form of phosphatidylinositol 3-kinase (PI3K) mimicked the activatory effects of glucose on PPI promoter activity, whereas Deltap85, a dominant negative form of the p85 subunit lacking the p110-binding domain, and the PI3K inhibitor LY 294002, blocked these effects. Similarly, glucose-stimulated nuclear trans-location of endogenous PDX-1 was blocked by Deltap85 expression, and wortmannin or LY 294002 blocked the trans-location from the nuclear membrane to the nucleoplasm of epitope-tagged PDX-1.c-myc. By contrast, SB 203580, an inhibitor of stress activated protein kinase-2 (SAPK2)/p38 MAP kinase, had no effect on any of the above parameters, and PPI promoter activity and PDX-1.c-myc localization were unaffected by over-expression of the upstream kinase MKK6 (MAP kinase kinase-6) or wild-type p38/SAPK2, respectively. Furthermore, no change in the activity of extracted p38/SAPK2 could be detected after incubation of cells at either 3 or 30 mm glucose. These data suggest that stimulation of PI3K is necessary and sufficient for the effects of glucose on PPI gene transcription, acting via a downstream signaling pathway that does not involve p38/SAPK2. PMID- 10821852 TI - SOCS-3 is an insulin-induced negative regulator of insulin signaling. AB - The SOCS proteins are induced by several cytokines and are involved in negative feedback loops. Here we demonstrate that in 3T3-L1 adipocytes, insulin, a hormone whose receptor does not belong to the cytokine receptor family, induces SOCS-3 expression but not CIS or SOCS-2. Using transfection of COS-7 cells, we show that insulin induction of SOCS-3 is enhanced upon Stat5B expression. Moreover, Stat5B from insulin-stimulated cells binds directly to a Stat element present in the SOCS-3 promoter. Once induced, SOCS-3 inhibits insulin activation of Stat5B without modifying the insulin receptor tyrosine kinase activity. Two pieces of evidence suggest that this negative regulation likely results from competition between SOCS-3 and Stat5B binding to the same insulin receptor motif. First, using a yeast two-hybrid system, we show that SOCS-3 binds to the insulin receptor at phosphotyrosine 960, which is precisely where Stat5B binds. Second, using confocal microscopy, we show that insulin induces translocation of SOCS-3 from an intracellular compartment to the cell membrane, leading to colocalization of SOCS-3 with the insulin receptor. This colocalization is dependent upon phosphorylation of insulin receptor tyrosine 960. Indeed, in cells expressing an insulin receptor mutant in which tyrosine 960 has been mutated to phenylalanine, insulin does not modify the cellular localization of SOCS-3. We have thus revealed an insulin target gene of which the expression is potentiated upon Stat5B activation. By inhibiting insulin-stimulated Stat5B, SOCS-3 appears to function as a negative regulator of insulin signaling. PMID- 10821853 TI - Differential regulation of exonic regulatory elements for muscle-specific alternative splicing during myogenesis and cardiogenesis. AB - Muscle-specific isoform of the mitochondrial ATP synthase gamma subunit (F(1)gamma) was generated by alternative splicing, and exon 9 of the gene was found to be lacking particularly in skeletal muscle and heart tissue. Recently, we reported that alternative splicing of exon 9 was induced by low serum or acidic media in mouse myoblasts, and that this splicing required de novo protein synthesis of a negative regulatory factor (Ichida, M., Endo, H., Ikeda, U., Matsuda, C., Ueno, E., Shimada, K., and Kagawa, Y. (1998) J. Biol. Chem. 273, 8492-8501; Hayakawa, M., Endo, H., Hamamoto, T., and Kagawa, Y. (1998) Biochem. Biophys. Res. Commun. 251, 603-608). In the present report, we identified a cis acting element on the muscle-specific alternatively spliced exon of F(1)gamma gene by an in vivo splicing system using cultured cells and transgenic mice. We constructed a F(1)gamma wild-type minigene, containing the full-length gene from exon 8 to exon 10, and two mutants; one mutant involved a pyrimidine-rich substitution on exon 9, whereas the other was a purine-rich substitution, abbreviated as F(1)gamma Pu-del and F(1)gamma Pu-rich mutants, respectively. Based on an in vivo splicing assay using low serum- or acid-stimulated splicing induction system in mouse myoblasts, Pu-del mutation inhibited exon inclusion, indicating that a Pu-del mutation would disrupt an exonic splicing enhancer. On the other hand, the Pu-rich mutation blocked muscle-specific exon exclusion following both inductions. Next, we produced transgenic mice bearing both mutant minigenes and analyzed their splicing patterns in tissues. Based on an analysis of F(1)gamma Pu-del minigene transgenic mice, the purine nucleotide of this element was shown to be necessary for exon inclusion in non-muscle tissue. In contrast, analysis of F(1)gamma Pu-rich minigene mice revealed that the F(1)gamma Pu-rich mutant exon had been excluded from heart and skeletal muscle of these transgenic mice, despite the fact mutation of the exon inhibited muscle-specific exon exclusion in myotubes of early embryonic stage. These results suggested that the splicing regulatory mechanism underlying F(1)gamma pre-mRNA differed between myotubes and myofibers during myogenesis and cardiogenesis. PMID- 10821854 TI - The alpha 5 chain of type IV collagen is the target of IgG autoantibodies in a novel autoimmune disease with subepidermal blisters and renal insufficiency. AB - We describe a novel autoimmune disease characterized by severe subepidermal bullous eruptions and renal insufficiency with IgG autoantibodies directed against the NC1 domain of the alpha5(IV) collagen chain. In vivo deposits of IgG and C3 were found along the dermal-epidermal junction of skin lesions. The identity of the target antigen was determined by immunochemical analyses of candidate antigens using the patients' autoantibodies. The patients' IgG autoantibodies reacted with a 185-kDa polypeptide that was distinguished from the known autoantigens of the extracellular matrix including type XVII collagen, type VII collagen, or the alpha3, beta3, and gamma2 chains of laminin 5. Preincubation of the serum with recombinant alpha5(IV)NC1 domain of type IV collagen abolished immunoreactivity with the 185-kDa antigen. The serum reacted specifically with the alpha5(IV)NC1, among the six NC1 domains of type IV collagen, by Western blot and enzyme-linked immunosorbent assay analyses. The patients' autoantibodies reacted with normal skin and renal glomerulus but not with skin and glomerulus of a patient with Alport syndrome in which the basement membranes are devoid of the alpha5(IV) collagen chain. This study provided for the first time unambiguous evidence for the alpha5(IV) collagen chain as the target antigen in a novel autoimmune disease characterized by skin and renal involvement. PMID- 10821855 TI - Potent and selective nonpeptide inhibitors of caspases 3 and 7 inhibit apoptosis and maintain cell functionality. AB - Caspases have been strongly implicated to play an essential role in apoptosis. A critical question regarding the role(s) of these proteases is whether selective inhibition of an effector caspase(s) will prevent cell death. We have identified potent and selective non-peptide inhibitors of the effector caspases 3 and 7. The inhibition of apoptosis and maintenance of cell functionality with a caspase 3/7 selective inhibitor is demonstrated for the first time, and suggests that targeting these two caspases alone is sufficient for blocking apoptosis. Furthermore, an x-ray co-crystal structure of the complex between recombinant human caspase 3 and an isatin sulfonamide inhibitor has been solved to 2.8-A resolution. In contrast to previously reported peptide-based caspase inhibitors, the isatin sulfonamides derive their selectivity for caspases 3 and 7 by interacting primarily with the S(2) subsite, and do not bind in the caspase primary aspartic acid binding pocket (S(1)). These inhibitors blocked apoptosis in murine bone marrow neutrophils and human chondrocytes. Furthermore, in camptothecin-induced chondrocyte apoptosis, cell functionality as measured by type II collagen promoter activity is maintained, an activity considered essential for cartilage homeostasis. These data suggest that inhibiting chondrocyte cell death with a caspase 3/7-selective inhibitor may provide a novel therapeutic approach for the prevention and treatment of osteoarthritis, or other disease states characterized by excessive apoptosis. PMID- 10821856 TI - Transcription factor Nrf2 coordinately regulates a group of oxidative stress inducible genes in macrophages. AB - Electrophiles and reactive oxygen species have been implicated in the pathogenesis of many diseases. Transcription factor Nrf2 was recently identified as a general regulator of one defense mechanism against such havoc. Nrf2 regulates the inducible expression of a group of detoxication enzymes, such as glutathione S-transferase and NAD(P)H:quinone oxidoreductase, via antioxidant response elements. Using peritoneal macrophages from Nrf2-deficient mice, we show here that Nrf2 also controls the expression of a group of electrophile- and oxidative stress-inducible proteins and activities, which includes heme oxygenase 1, A170, peroxiredoxin MSP23, and cystine membrane transport (system x(c)(-)) activity. The response to electrophilic and reactive oxygen species-producing agents was profoundly impaired in Nrf2-deficient cells. The lack of induction of system x(c)(-) activity resulted in the minimum level of intracellular glutathione, and Nrf2-deficient cells were more sensitive to toxic electrophiles. Several stress agents induced the DNA binding activity of Nrf2 in the nucleus without increasing its mRNA level. Thus Nrf2 regulates a wide-ranging metabolic response to oxidative stress. PMID- 10821857 TI - Nuclear NonO/p54(nrb) protein is a nonclassical carbonic anhydrase. AB - The growing carbonic anhydrase (CA) gene family includes 11 enzymatically active isozymes in mammals. Each of them has a characteristic cellular and subcellular distribution pattern. In this report, we demonstrate for the first time a nuclear protein with CA activity. A polypeptide recognized by CA II antibodies was purified from several rat tissues using CA inhibitor affinity chromatography. This polypeptide of apparent 66 kDa mass was characterized using amino acid sequencing and CA activity measurements. It appeared to be identical to nonO/p54(nrb), a previously cloned and characterized RNA and DNA binding nuclear factor. Recombinant nonO generated in baculovirus bound to the CA inhibitor affinity chromatography matrix and revealed detectable CA activity (25 units/mg). Hansson's histochemical staining of rat lymph nodes followed by light and electron microscopy showed nuclear CA activity in lymphocytes, suggesting that the nuclear nonO protein is catalytically active in vivo. These results demonstrate that a previously known transcription factor is a novel, nonclassical CA. Through its CA activity, the nonO may function in the maintenance of pH homeostasis in the nucleus. PMID- 10821858 TI - Crystal structures of an oxygen-binding cytochrome c from Rhodobacter sphaeroides. AB - The photosynthetic bacterium Rhodobacter sphaeroides produces a heme protein (SHP), which is an unusual c-type cytochrome capable of transiently binding oxygen during autooxidation. Similar proteins have not only been observed in other photosynthetic bacteria but also in the obligate methylotroph Methylophilus methylotrophus and the metal reducing bacterium Shewanella putrefaciens. A three dimensional structure of SHP was derived using the multiple isomorphous replacement phasing method. Besides a model for the oxidized state (to 1.82 A resolution), models for the reduced state (2.1 A resolution), the oxidized molecule liganded with cyanide (1. 90 A resolution), and the reduced molecule liganded with nitric oxide (2.20 A resolution) could be derived. The SHP structure represents a new variation of the class I cytochrome c fold. The oxidized state reveals a novel sixth heme ligand, Asn(88), which moves away from the iron upon reduction or when small molecules bind. The distal side of the heme has a striking resemblance to other heme proteins that bind gaseous compounds. In SHP the liberated amide group of Asn(88) stabilizes solvent-shielded ligands through a hydrogen bond. PMID- 10821859 TI - The structure and the characteristic DNA binding property of the C-terminal domain of the RNA polymerase alpha subunit from Thermus thermophilus. AB - The C-terminal domain of the alpha subunit of the RNA polymerase (alphaCTD) from Escherichia coli (Ec) regulates transcription by interacting with many kinds of proteins and promoter upstream (UP) elements consisting of AT-rich sequences. However, it is unclear how this system is common in all eubacteria. We investigate the structure and properties of alphaCTD from an extremely thermophilic eubacterium, Thermus thermophilus (Tt). The solution structure of Tt alphaCTD (85 amino acids) was determined by NMR, and the interaction between Tt alphaCTD and DNA with different sequences was investigated by means of chemical shift perturbation experiments. The tertiary structure of Tt alphaCTD is almost identical with that of Ec alphaCTD despite 32% sequence homology. However, Tt alphaCTD interacts with the upstream region sequence of the promoter in the Tt 16 S ribosomal protein operon rather than the Ec UP element DNA. The upstream region sequence of Tt is composed of 25 base pairs with 40% AT, unlike the Ec UP element with 80% AT. The DNA binding site in Tt alphaCTD is located on the surface composed of helix 4 and the loop preceding helix 4. The electric charges on this surface are not remarkably localized like those of Ec alphaCTD. PMID- 10821860 TI - RecA force generation by hydrolysis waves. AB - We present a simple theory of the dynamics of force generation by RecA during homologous strand exchange and a continuous, deterministic mathematical model of the proposed process. Calculations show that force generation is possible in this model for certain reasonable values of the parameters. We predict the shape of the force-velocity curve for the Holliday junction, which exhibits a distinctive kink at large retarding force, and suggest experiments which should distinguish between the proposed model and other models in the literature. PMID- 10821861 TI - Fibroblast growth factor (FGF) receptor 1-IIIb is a naturally occurring functional receptor for FGFs that is preferentially expressed in the skin and the brain. AB - Fibroblast growth factors (FGFs) transmit their signals through four transmembrane receptors that are designated FGFR1-4. Alternative splicing in the extracellular region of FGFR1-3 generates receptor variants with different ligand binding affinities. Thus two types of transmembrane receptors (IIIb and IIIc isoforms) have been identified for FGFR2 and FGFR3, and the existence of analogous variants has been postulated for FGFR1 based on its genomic structure. However, only a single full-length transmembrane FGFR1 variant (FGFR1-IIIc) has been identified so far. Here we describe the cloning of a full-length cDNA encoding FGFR1-IIIb from a mouse skin wound cDNA library. This receptor isoform was expressed at the highest levels in a subset of sebaceous glands of the skin and in neurons of the hippocampus and the cerebellum. FGFR1-IIIb was expressed in L6 rat skeletal muscle myoblasts and used in cross-linking and receptor binding studies. FGF-1 was found to bind the receptor with high affinity, whereas FGF-2, 10, and -7 bound with significantly lower affinities. Despite their apparently similar but low affinities, FGF-10 but not FGF-7 induced the activation of p44/42 mitogen-activated protein kinase in FGFR1-IIIb-expressing L6 myoblasts and stimulated mitogenesis in these cells, demonstrating that this new receptor variant is a functional transmembrane receptor for FGF-10. PMID- 10821862 TI - Structural and functional consequences of peptide-carbohydrate mimicry. Crystal structure of a carbohydrate-mimicking peptide bound to concanavalin A. AB - The functional consequences of peptide-carbohydrate mimicry were analyzed on the basis of the crystal structure of concanavalin A (ConA) in complex with a carbohydrate-mimicking peptide, DVFYPYPYASGS. The peptide binds to the non crystallographically related monomers of two independent dimers of ConA in two different modes, in slightly different conformations, demonstrating structural adaptability in ConA-peptide recognition. In one mode, the peptide has maximum interactions with ConA, and in the other, it shows relatively fewer contacts within this site but significant contacts with the symmetry-related subunit. Neither of the peptide binding sites overlaps with the structurally characterized mannose and trimannose binding sites on ConA. Despite this, the functional mimicry between the peptide and carbohydrate ligands was evident. The peptide inhibited ConA induced T cell proliferation in a dose-dependent manner. The effect of the designed analogs of the peptide on ConA-induced T cell proliferation and their recognition by the antibody response against alpha-d mannopyranoside indicate a role for aromatic residues in functional mimicry. Although the functional mimicry was observed between the peptide and carbohydrate moieties, the crystal structure of the ConA-peptide complex revealed that the two peptide binding sites are independent of the methyl alpha-d-mannopyranoside binding site. PMID- 10821863 TI - Muscle differentiation is antagonized by SOX15, a new member of the SOX protein family. AB - SOX proteins belong to a multigenic family characterized by a unique DNA binding domain, known as the high mobility group box, that is related to that of the testis determining gene SRY. cDNA sequences for more than 30 SOX genes have been identified, and some are known to have diverse roles in vertebrate differentiation and development. Here, we report the isolation and characterization of mouse Sox15 that was uncovered during a screen for high mobility group box containing transcription factors that are expressed at different levels during skeletal muscle differentiation. Sox15 cDNAs were found at a much higher frequency in myoblasts prior to their differentiation into myotubes. Electrophoretic mobility shift assays indicated that recombinant SOX15 protein was capable of binding to a consensus DNA binding site for SOX proteins. When overexpressed in C2C12 myoblasts, wild type SOX15, but not a C-terminal truncated form or the related protein SOX11, specifically inhibited activation of muscle-specific genes and expression of the basic helix-loop-helix myogenic factors myogenin and MyoD, resulting in a failure of the cells to differentiate into myotubes. These results suggest a specific and repressive role for SOX15, requiring the C-terminal domain, during myogenesis. PMID- 10821864 TI - Determinants of cytochrome c pro-apoptotic activity. The role of lysine 72 trimethylation. AB - Cytochrome c released from vertebrate mitochondria engages apoptosis by triggering caspase activation. We previously reported that, whereas cytochromes c from higher eukaryotes can activate caspases in Xenopus egg and mammalian cytosols, iso-1 and iso-2 cytochromes c from the yeast Saccharomyces cerevisiae cannot. Here we examine whether the inactivity of the yeast isoforms is related to a post-translational modification of lysine 72, N-epsilon-trimethylation. This modification was found to abrogate pro-apoptotic activity of metazoan cytochrome c expressed in yeast. However, iso-1 cytochrome c lacking the trimethylation modification also was devoid of pro-apoptotic activity. Thus, both lysine 72 trimethylation and other features of the iso-1 sequence preclude pro-apoptotic activity. Competition studies suggest that the lack of pro-apoptotic activity was associated with a low affinity for Apaf-1. As cytochromes c that lack apoptotic function still support respiration, different mechanisms appear to be involved in the two activities. PMID- 10821865 TI - Movement of the biotin carboxylase B-domain as a result of ATP binding. AB - Acetyl-CoA carboxylase catalyzes the first committed step in fatty acid synthesis. In Escherichia coli, the enzyme is composed of three distinct protein components: biotin carboxylase, biotin carboxyl carrier protein, and carboxyltransferase. The biotin carboxylase component has served for many years as a paradigm for mechanistic studies devoted toward understanding more complicated biotin-dependent carboxylases. The three-dimensional x-ray structure of an unliganded form of E. coli biotin carboxylase was originally solved in 1994 to 2.4-A resolution. This study revealed the architecture of the enzyme and demonstrated that the protein belongs to the ATP-grasp superfamily. Here we describe the three-dimensional structure of the E. coli biotin carboxylase complexed with ATP and determined to 2.5-A resolution. The major conformational change that occurs upon nucleotide binding is a rotation of approximately 45(o) of one domain relative to the other domains thereby closing off the active site pocket. Key residues involved in binding the nucleotide to the protein include Lys-116, His-236, and Glu-201. The backbone amide groups of Gly-165 and Gly-166 participate in hydrogen bonding interactions with the phosphoryl oxygens of the nucleotide. A comparison of this closed form of biotin carboxylase with carbamoyl phosphate synthetase is presented. PMID- 10821866 TI - Death signal-induced localization of p53 protein to mitochondria. A potential role in apoptotic signaling. AB - The mechanism of p53-mediated apoptosis after cellular stress remains poorly understood. Evidence suggests that p53 induces cell death by a multitude of molecular pathways involving activation of target genes and transcriptionally independent direct signaling. Mitochondria play a key role in apoptosis. We show here that a fraction of p53 protein localizes to mitochondria at the onset of p53 dependent apoptosis but not during p53-independent apoptosis or p53-mediated cell cycle arrest. The accumulation of p53 to mitochondria is rapid (within 1 h after p53 activation) and precedes changes in mitochondrial membrane potential, cytochrome c release, and procaspase-3 activation. Immunoelectron microscopy and immuno-fluorescence-activated cell sorter analysis of isolated mitochondria show that the majority of mitochondrial p53 localizes to the membranous compartment, whereas a fraction is found in a complex with the mitochondrial import motor mt hsp70. After induction of ectopic p53 without additional DNA damage in p53 deficient cells, p53 again partially localizes to mitochondria, preceding the onset of apoptosis. Overexpression of anti-apoptotic Bcl-2 or Bcl-xL abrogates stress signal-mediated mitochondrial p53 accumulation and apoptosis but not cell cycle arrest, suggesting a feedback signaling loop between p53 and mitochondrial apoptotic regulators. Importantly, bypassing the nucleus by targeting p53 to mitochondria using import leader fusions is sufficient to induce apoptosis in p53 deficient cells. We propose a model where p53 can contribute to apoptosis by direct signaling at the mitochondria, thereby amplifying the transcription dependent apoptosis of p53. PMID- 10821867 TI - Site-selective dephosphorylation of the platelet-derived growth factor beta receptor by the receptor-like protein-tyrosine phosphatase DEP-1. AB - Ligand stimulation of PDGF beta-receptors leads to autophosphorylation of the regulatory tyrosine 857 and of tyrosine residues that in their phosphorylated form serve as docking sites for Src homology 2 domain-containing proteins. Regulation of the PDGF beta-receptor by protein-tyrosine phosphatases is poorly understood. We have investigated PDGF beta-receptor dephosphorylation by receptor like protein-tyrosine phosphatase DEP-1 using a cell line with inducible DEP-1 expression and by characterizing in vitro dephosphorylation of the PDGF beta receptor and of receptor-derived phosphopeptides by DEP-1. After DEP-1 induction PDGF beta-receptor.DEP-1 complexes and reduced receptor tyrosine phosphorylation were observed. Phosphopeptide analysis of the PDGF beta-receptors from DEP-1 expressing cells and of the receptors dephosphorylated in vitro by DEP-1 demonstrated that dephosphorylation of autophosphorylation sites of the receptor differed and revealed that the regulatory Tyr(P)(857) was not a preferred site for DEP-1 dephosphorylation. When dephosphorylation of synthetic receptor-derived peptides was analyzed, the selectivity was reproduced, indicating that amino acid sequence surrounding the phosphorylation sites is the major determinant of selectivity. This notion is supported by the observation that the poorly dephosphorylated Tyr(P)(562) and Tyr(P)(857), in contrast to other analyzed phosphorylation sites, are surrounded by basic amino acid residues at positions 4 and +3 relative to the tyrosine residue. Our study demonstrates that DEP-1 dephosphorylation of the PDGF beta-receptor is site-selective and may lead to modulation, rather than general attenuation, of signaling. PMID- 10821868 TI - GLUT8, a novel member of the sugar transport facilitator family with glucose transport activity. AB - GLUT8 is a novel glucose transporter-like protein that exhibits significant sequence similarity with the members of the sugar transport facilitator family (29.4% of amino acids identical with GLUT1). Human and mouse sequence (86.2% identical amino acids) comprise 12 putative membrane-spanning helices and several conserved motifs (sugar transporter signatures), which have previously been shown to be essential for transport activity, e.g. GRK in loop 2, PETPR in loop 6, QQLSGVN in helix 7, DRAGRR in loop 8, GWGPIPW in helix 10, and PETKG in the C terminal tail. An expressed sequence tag (STS A005N15) corresponding with the 3' untranslated region of GLUT8 has previously been mapped to human chromosome 9. COS-7 cells transfected with GLUT8 cDNA expressed a 42-kDa protein exhibiting specific, glucose-inhibitable cytochalasin B binding (K(D) = 56.6 +/- 18 nm) and reconstitutable glucose transport activity (8.1 +/- 1. 4 nmol/(mg protein x 10 s) versus 1.1 +/- 0.1 in control transfections). In human tissues, a 2.4-kilobase pair transcript was predominantly found in testis, but not in testicular carcinoma. Lower amounts of the mRNA were detected in most other tissues including skeletal muscle, heart, small intestine, and brain. GLUT8 mRNA was found in testis from adult, but not from prepubertal rats; its expression in human testis was suppressed by estrogen treatment. It is concluded that GLUT8 is a sugar transport facilitator with glucose transport activity and a hormonally regulated testicular function. PMID- 10821869 TI - Live Salmonella modulate expression of Rab proteins to persist in a specialized compartment and escape transport to lysosomes. AB - We investigated the intracellular route of Salmonella in macrophages to determine a plausible mechanism for their survival in phagocytes. Western blot analysis of isolated phagosomes using specific antibodies revealed that by 5 min after internalization dead Salmonella-containing phagosomes acquire transferrin receptors (a marker for early endosomes), whereas by 30 min the dead bacteria are found in vesicles carrying the late endosomal markers cation-dependent mannose 6 phosphate receptors, Rab7 and Rab9. In contrast, live Salmonella-containing phagosomes (LSP) retain a significant amount of Rab5 and transferrin receptor until 30 min, selectively deplete Rab7 and Rab9, and never acquire mannose 6 phosphate receptors even 90 min after internalization. Retention of Rab5 and Rab18 and selective depletion of Rab7 and Rab9 presumably enable the LSP to avoid transport to lysosomes through late endosomes. The presence of immature cathepsin D (48 kDa) and selective depletion of the vacuolar ATPase in LSP presumably contributes to the less acidic pH of LSP. In contrast, proteolytically processed cathepsin D (M(r) 17,000) was detected by 30 min on the dead Salmonella containing phagosomes. Morphological analysis also revealed that after uptake by macrophages, the dead Salmonella are transported to lysosomes, whereas the live bacteria persist in compartments that avoid fusion with lysosomes, indicating that live Salmonella bypass the normal endocytic route targeted to lysosomes and mature in a specialized compartment. PMID- 10821870 TI - Glutathione redox potential modulated by reactive oxygen species regulates translation of Rubisco large subunit in the chloroplast. AB - Previous work showed a transient but dramatic arrest in the synthesis of Rubisco large subunit (LSU) upon transfer of Chlamydomonas reinhardtii cells from low light (LL) to high light (HL). Using dichlorofluorescin, a short-term increase in reactive oxygen species (ROS) was demonstrated, suggesting that their excessive formation could signal LSU down-regulation. A decrease in LSU synthesis occurred at LL in the presence of methyl viologen and was prevented at HL by ascorbate. Interfering with D1 function by mutations or by incubation with DCMU prevented the increase in ROS formation at HL and the concomitant down-regulation of LSU synthesis. If the electron transport was blocked further downstream, by mutation in the cytochrome b(6)/f or by incubation with 2, 5-dibromo-3-methyl-6-isopropyl p-benzoquinone, ROS formation increased, and LSU synthesis ceased. The elevation of ROS occurred concurrently with a change in the redox state of the glutathione pool, which shifted toward its oxidized form immediately after the transfer to HL and returned to its original value after 6 h. The decrease in the reduced/oxidized glutathione ratio at HL was prevented by ascorbate and could be induced at LL by methyl viologen. We suggest that excess ROS mediate a decrease in the reduced/oxidized glutathione ratio that in turn signals the translational arrest of the rbcL transcript. PMID- 10821871 TI - Mitochondria of Saccharomyces cerevisiae contain one-conserved cysteine type peroxiredoxin with thioredoxin peroxidase activity. AB - Peroxiredoxins are ubiquitously expressed proteins that reduce hydroperoxides using disulfur-reducing compounds as electron donors. Peroxiredoxins (Prxs) have been classified in two groups dependent on the presence of either one (1-Cys Prx) or two (2-Cys Prx) conserved cysteine residues. Moreover, 2-Cys Prxs, also named thioredoxin peroxidases, have peroxide reductase activity with the use of thioredoxin as biological electron donor. However, the biological reducing agent for the 1-Cys Prx has not yet been identified. We report here the characterization of a 1-Cys Prx from yeast Saccharomyces cerevisiae that we have named Prx1p. Prx1p is located in mitochondria, and it is overexpressed when cells use the respiratory pathway, as well as in response to oxidative stress conditions. We show also that Prx1p has peroxide reductase activity in vitro using the yeast mitochondrial thioredoxin system as electron donor. In addition, a mutated form of Prx1p containing the absolutely conserved cysteine as the only cysteine residue also shows thioredoxin-dependent peroxide reductase activity. This is the first example of 1-Cys Prx that has thioredoxin peroxidase activity. Finally, exposure of null Prx1p mutant cells to oxidant conditions reveals an important role of the mitochondrial 1-Cys Prx in protection against oxidative stress. PMID- 10821872 TI - Anti-apoptotic role of focal adhesion kinase (FAK). Induction of inhibitor-of apoptosis proteins and apoptosis suppression by the overexpression of FAK in a human leukemic cell line, HL-60. AB - Focal adhesion kinase (FAK) has an anti-apoptotic role in anchorage-dependent cells via an unknown mechanism. To elucidate the role of FAK in anti-apoptosis, we have established several FAK cDNA-transfected HL-60 cell lines and examined whether FAK-transfected cells have resistance to apoptotic stimuli. FAK transfected HL-60 (HL-60/FAK) cells were highly resistant to apoptosis induced with hydrogen peroxide (1 mm) and etoposide (50 microg/ml) compared with the parental HL-60 cells or the vector-transfected cells, when determined using viability assay, DNA fragmentation, and flow cytometry analysis. Because no proteolytic cleavage of pro-caspase 3 to mature caspase 3 fragment was observed in HL-60/FAK cells, FAK was presumed to inhibit an upstream signal pathway leading to the activation of caspase 3. HL-60/FAK activated the phosphatidylinositide 3'-OH-kinase-Akt survival pathway and exhibited significant activation of NF-kappaB with marked induction of inhibitor-of-apoptosis proteins (IAPs: cIAP-1, cIAP-2, XIAP), regardless of the hydrogen peroxide-treated or untreated conditions, whereas no significant IAPs were detected in the parental or vector-transfected HL-60 cells. Apoptotic agents induced higher NF-kappaB activation in HL-60/FAK cells than in HL-60/Vect cells, and it appeared that sustained NF-kappaB activation is critical to the anti-apoptotic states in HL 60/FAK cells. Mutagenesis of FAK cDNA revealed that Y397 and Y925, which are involved in the tyrosine-phosphorylation sites, were prerequisite for the anti apoptotic activity as well as induction of IAPs, and that K454, which is involved in the kinase activity, was also required for the full anti-apoptotic activity of FAK. Taken together, we have demonstrated definitively that FAK-transfected HL-60 cells, otherwise sensitive to apoptosis, become resistant to the apoptotic stimuli. We conclude that FAK activates the phosphatidylinositide 3'-OH-kinase Akt survival pathway with the concomitant activation of NF-kB and induction of IAPs, which ultimately inhibit apoptosis by inhibiting caspase-3 cascade. PMID- 10821873 TI - High intensity resistance training: effects on bone in older men and women. AB - There is evidence that high intensity resistance training promotes bone maintenance in older women, however, the effect of high intensity free weight training has not been investigated in older men or women. Furthermore, little is known about the chronic effect of weight training on serum insulin growth factor I (IGF-I) in this population. We compared the effects of a moderate intensity seated resistance-training program with a high intensity standing free weight exercise program on bone mass and serum levels of IGF-I and IGFBP3 in healthy older men and women. Twenty-eight men (54.6 +/- 3. 2 years) and 26 nonestrogen replaced women (52.8 +/- 3.3 years) served as their own controls for 12 weeks, then were randomly assigned to a moderate or high intensity training group and trained three times/week for 24 weeks. Prior to and after the control period and at the end of training, bone mass and body composition were assessed by dual energy X-ray absorptiometry (DXA), muscle strength by isokinetic dynamometry, muscular power by Wingate Anaerobic Power Test, and IGF-I by radioimmunoassay (RIA). A repeated measures analysis of covariance (ANCOVA) revealed that high intensity training resulted in a gain in spine BMD in men (1.9%), P < 0.05, but not in women, whereas moderate intensity training produced no changes in either gender at this site. Increases were observed at the greater trochanter, P < 0.03, in men regardless of training intensity, but not in women at any hip site. However, when compared with zero, both men and women in the high intensity group demonstrated significant increases in trochanteric BMD (1.3% and 2. 0%, respectively) and a decrease in femoral BMD (-1.8%). Neither circulating serum IGF-I nor IGFBP3 were altered by either training regimen, but both training programs resulted in improvements in total body strength (37.62%) and lean mass (males 4.1%, females 3. 1%). We conclude that although resistance training of moderate to high intensity produced similar muscle changes in older adults, a higher magnitude is necessary to stimulate osteogenesis at the spine. However, at the spine, intensity was not sufficient to offset low levels of estrogen in early postmenopausal women. Furthermore, bone changes were not accompanied by changes in circulating serum levels of IGF-I or IGFBP3. PMID- 10821874 TI - PvuII polymorphisms of the estrogen receptor alpha and bone mineral density in healthy southern Chinese women. AB - The association between PvuII polymorphisms of the estrogen receptor alpha (ERalpha) gene and total as well as regional bone mineral density (BMD) in healthy Chinese women (n = 182) was determined by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP), where P indicated the absence and p the presence of PvuII restriction sites. Subjects with PP genotype had significantly higher BMD at the thoracic spine and ribs (both P < 0.05) when compared with those with Pp and pp genotypes. Although PP genotype had slightly higher BMD values at the lumbar spine L2-L4 region and hip by 8% and 7%, respectively, the results failed to reach statistical significance. After adjusting for age, height, weight, and years since menopause, PP genotype had higher BMD at the left (P < 0.02) and right (P < 0.05) rib region but not at the thoracic spine (P = 0.056). Analyzing the premenopausal subjects alone (n = 64) revealed that subjects with PP genotype had higher adjusted BMD at the right rib region (P < 0.05). When only the postmenopausal women (n = 118) were analyzed, the adjusted BMD of the PP genotype at the thoracic spine was significantly higher (P < 0.05) than the other two groups. In conclusion, estrogen receptor gene has a role in determining bone mass but the clinical impact on its own is probably small. PMID- 10821875 TI - Lack of influence of collagen type Ialpha1 Sp1 binding site polymorphism on the rate of bone loss in a cohort of postmenopausal danish women followed for 18 years. AB - A polymorphism in an Sp1 site in the collagen Ialpha1 (COLIA1) gene has recently been identified and the Ss and ss genotypes were shown to be potentially important determinants of low bone mass in postmenopausal women. Additionally, in a Dutch population, the association of the COLIA1 polymorphism with low bone mineral density (BMD) was more pronounced with increasing age, suggesting a genotype effect on the rate of bone loss. We have investigated the relationship between the COLIA1 Sp1 polymorphism and the rate of bone loss in a longitudinal study with a total of 133 postmenopausal women followed for 18 years. The frequencies of the genotypes were SS 70.7%, Ss 27.8%, ss 1.5% and were in Hardy Weinberg equilibrium. No association of the COLIA1 genotype with rate of bone loss was detected and there was no difference between the genotype groups with respect to BMD at the femoral neck or lumbar spine. Women with the Ss or ss genotypes, who have been postulated to have low BMD, had even higher BMD at the lower forearm than women with the SS genotype. The levels of serum osteocalcin and urinary collagen type I degradation products were not found to be associated with the COLIA1 Sp1 polymorphism. In conclusion, this study does not support the hypothesis that the Ss COLIA1 genotype predisposes women to increased rate of bone loss or low BMD. However, because of a low absolute number of the ss genotype, it was not possible to reach a conclusion on this particular genotype with regard to an association with low BMD or rate of bone loss. PMID- 10821876 TI - Effects of age on serum dehydroepiandrosterone sulfate, IGF-I, and IL-6 levels in women. AB - Data from animal and in vitro studies suggest that the growth-promoting effects of the adrenal androgen dehydroepiandrosterone sulfate (DHEAS) may be mediated by stimulation of insulin-like growth factor-I (IGF-I) and/or inhibition of interleukin 6 (IL-6), a cytokine mediator of bone resorption. This study tests the hypotheses that there are effects of age on serum DHEAS, IGF-I, and IL-6 levels, and that levels of IGF-I and IL-6 are related to DHEAS levels. The study included 102 women: 27 premenopausal and 75 postmenopausal, including 35 postmenopausal women with osteoporosis, as defined by bone mineral density scores by dual X-ray energy absorptiometry. DHEAS levels decreased significantly with age (r = -0.52, P < 0.0001) and IGF-I levels decreased significantly with age (r = -0.49, P < 0.0001). IL-6 levels increased significantly with age (r = 0.36, P = 0.008). IGF-I was positively correlated to DHEAS levels (r = 0.43, P < 0. 0001, n = 102) and IL-6 levels were negatively correlated to DHEAS levels (r = -0.32, P = 0.021, n = 54). Levels of DHEAS and IGF-I were correlated with T scores of the spine and some hip sites. In a multiple variable model to predict DHEAS, age was an important predictor (P < 0.001), but osteoporosis status, IGF-I, and IL-6 were not. The median DHEAS level was lower in the postmenopausal osteoporotic women (67 microg/dl, n = 35) than in the nonosteoporotic postmenopausal women (106.3 microg/dl, n = 40, P = 0. 03), but this was not significant after correction for age. Age accounted for 32% of the variance in DHEAS levels. In summary, DHEAS levels decreased with age and had a positive association with IGF-I levels and a negative association with IL-6 levels. DHEA deficiency may contribute to age related bone loss through anabolic (IGF-I) and anti-osteolytic (IL-6) mechanisms. PMID- 10821877 TI - Hypovitaminosis D myopathy without biochemical signs of osteomalacic bone involvement. AB - The aims of this study were to investigate myopathy in relation to vitamin D status, and to study the muscular effects of vitamin D treatment on vitamin D deficient individuals. Further, hypovitaminosis D myopathy was investigated in relation to alkaline phosphatase (ALP), the most commonly used marker for hypovitaminosis D osteopathy. Eight patients with osteomalacia had an isokinetic dynamometer test of all major muscle groups before and after 3 months of vitamin D treatment. The most pronounced improvements in muscle power were seen in the weight-bearing antigravity muscles of the lower limbs. A cross-sectional study was performed among 55 vitamin D-deficient veiled Arab women living in Denmark and 22 Danish controls. An isometric dynamometer model was used for determination of quadriceps muscle power. Both maximal voluntary contraction (MVC) and electrically stimulated values (single twitch, maximal production rate (MPR), and maximal relaxation rate (MRR)) were determined. The women underwent high-dose vitamin D treatment and were retested after 3 and 6 months. Prior to vitamin D treatment all parameters of muscle function in the group of vitamin D-deficient Arab women were significantly reduced compared with Danish controls. MVC: 259.4 +/- 11.0 N (Newton) versus 392.6 +/- 11. 4 N (P < 10(-6)), single twitch: 47.0 +/ 1.8 N versus 74.6 +/- 2.2 N (P < 10(-5)), MPR 8.9 +/- 0.3 N/10 ms versus 14.3 +/ 0.4 N/10 ms (P < 10(-6)), MRR 4.5 +/- 0.2 N/10 ms versus 6.2 +/- 0.2 N/10 ms (P < 10(-6)). Muscle function was affected to a similar degree in women with and without bone involvement (as indicated by elevated ALP). After 3 months of vitamin D treatment all muscle-related parameters improved significantly. After 6 months only MVC was reduced compared with Danish controls (320.7 +/- 14.3 N (P < 0.02)), whereas all other measurements were normalized. Hypovitaminosis D myopathy is a prominent symptom of vitamin D deficiency, and severely impaired muscle function may be present even before biochemical signs of bone disease develop. Full normalization of hypovitaminosis D myopathy demands high-dose vitamin D treatment for 6 months or more. Our findings indicate that serum levels of ALP cannot be used in the screening for hypovitaminosis D myopathy. Assessment of s-25OHD is the only reliable test. PMID- 10821878 TI - Calcium-induced natriuresis: physiologic and clinical implications. AB - Assessment of the tubular reabsorption of calcium (Ca) by infusion is complicated by suppression of parathyroid hormone (PTH) secretion, and by activation of the serpentine Ca sensing receptor in the renal tubule, which inhibits Ca and sodium reabsorption, but little is known about the magnitude of the natriuretic effect of Ca in human subjects. Accordingly, we reanalyzed the relationship between serum Ca and urine Ca and sodium excretion expressed per unit of creatinine clearance (CaE and NaE), and per unit of time (UCa and UNa), during a standard Ca infusion, in 14 healthy volunteers and in 8 primary hyperparathyroid patients. In healthy subjects we observed a large effect of Ca infusion on NaE, which rose as high as 8 mmol/liter GFR. In patients with primary hyperparathyroidism both CaE and NaE during Ca infusion were significantly lower overall than in healthy subjects for comparable values of serum Ca (P < 0.05 by covariance analysis), due mainly to a decline or reversal of the slopes at the highest serum Ca levels. In both controls and primary hyperparathyroid subjects the variance of CaE as dependent variable was explained by both serum Ca and by NaE as independent variables (P < 0.001). We conclude that (1) The natriuretic effect of hypercalcemia was unexpected large and if maintained would lead to substantial depletion of extracellular fluid. (2) Patients with chronic hypercalcemia, including primary hyperparathyroidism, probably have mild sodium depletion, and are more susceptible to volume depletion. (3) Calcium reabsorption during Ca infusion is reduced by suppression of PTH secretion and increased by volume contraction due to sodium depletion. Discrimination between different basal levels of parathyroid function is successful because these effects usually cancel out. (4) The increase in tubular reabsorption of Ca due to volume contraction can initiate a vicious circle, of importance to the pathogenesis and treatment of severe hypercalcemia. PMID- 10821879 TI - Matrix vesicle calcification in bones of adult rats. AB - To clarify the calcification mechanism that functions in bone formation in adult rats, the ultrastructure of tibial trabeculae and calvarial endostea obtained from 8- to 18-month-old rats was investigated morphologically, and compared with that of 19.5-day post-coitum fetal rats. In both samples, osteoid was observed between the activated osteoblasts and the calcified matrix, which contained matrix vesicles enclosed by a biological membrane. Some of these vesicles contained needle-like crystals thought to be hydroxyapatite, suggesting probable matrix vesicle calcification. These results indicate that matrix vesicle function not only in the initial calcification that occurs during embryonic ossification but also contribute to bone formation in adults. PMID- 10821880 TI - Development and application of a serum C-telopeptide and osteocalcin assay to measure bone turnover in an ovariectomized rat model. AB - Biochemical markers applicable to the ovariectomized rat model can provide important tools for studying the bone remodeling process in this animal model of postmenopausal osteoporosis. We describe the development and application of two biochemical markers, a C-telopeptide (of type-I collagen) enzyme-linked immunosorbent assay (ELISA) for measuring bone resorption and an osteocalcin radioimmunoassay (RIA) for measuring bone formation in rat serum. The C telopeptide ELISA is based on an affinity purified polyclonal antibody generated against human sequence DFSFLPQPPQEKAHDGGR. The antibody epitope involves amino acid sequence, which is similar in rat and human carboxyl terminal peptide of type-I (alpha 1) collagen. Sensitivity of the ELISA was 0.3 ng/ml. The averaged intra- and interassay variation was CV <7%. Averaged dilution and spiked recoveries were 91% and 105%, respectively. The second marker developed is a synthetic peptide-based osteocalcin RIA, which does not require isolation and purification of intact osteocalcin from rat bone. Osteocalcin antiserum used in the RIA was generated in rabbits against a synthetic peptide comprising amino acids 33-49 of the rat osteocalcin sequence. The sensitivity of the RIA was 0.15 ng/ml of peptide. The averaged intra (n = 10) and interassay variations for two controls were CV <9% and 12%, respectively. The averaged dilution and spiked recoveries were 99.6%. In vivo validation of the C-telopeptide ELISA and osteocalcin RIA was performed in an ovariectomized (OVX) rat model. In 12-week old OVX Sprague Dawley rats, the C-telopeptide and osteocalcin concentrations were approximately 65% and 40%, respectively, higher than the sham group. Estradiol repletion significantly lowered the C-telopeptide and osteocalcin concentration to the levels of the sham group. In addition, changes in serum C telopeptide concentration correlated negatively with trabecular BMD measured by pQCT (r = -0.51, P < 0.001). In conclusion, the C-telopeptide ELISA and osteocalcin RIA exhibited required sensitivity, accuracy, and adequate discriminatory power to be used for measuring bone resorption and bone formation in the ovariectomized rat model. PMID- 10821881 TI - Dietary fish oil supplementation adversely affects cortical bone morphology and biomechanics in growing rabbits. AB - Despite substantial evidence that fish oil-derived (n-3) polyunsaturated fatty acids (PUFA) may protect against cardiovascular disease, the effects of supplements containing (n-3) PUFA on the skeletal system are unknown. Here we investigated how a diet supplemented with 10 g/100 g fish oil affected tibial cortical morphology and mechanical properties in weanling rabbits. Rabbits were subdivided into a normal control (n = 10), a fish oil (n = 20), and a pair-fed (n = 20) group. The pair-fed group was energy restricted to match average body mass of the fish oil group. At completion of the 40 day dietary intervention, control rabbits were significantly heavier than the other two groups. Comparison between control and pair-fed rabbits revealed that energy restriction alone (30%) did not induce significant changes in tibial middiaphyseal morphology, but tibial longitudinal growth was significantly impaired. Most tibial mechanical properties were significantly degraded by energy restriction. Fish oil-supplemented rabbits had significantly smaller middiaphyseal areal properties and shorter tibiae than pair-fed rabbits. Tibial structural properties were significantly reduced in fish oil-fed rabbits, but tibial stress at the proportional limit (material property) was not significantly affected. Our data suggest that 10% fish oil supplementation in the presence of modest vitamin E supplementation can have detrimental effects on the skeleton of rapidly growing rabbits. PMID- 10821882 TI - Anionic effects on the size and shape of apatite crystals grown from physiological solutions. AB - Comparatively little is known of the role tissue fluid electrolytes have in establishing the size and shape of apatite crystals deposited in skeletal tissues. In vitro accretion experiments using synthetic apatite seed crystals comparable in size to bone apatite were performed to assess the extent to which these crystalline features may be affected by direct electrolyte/mineral interactions. A constant composition method was used to maintain the accretion reactions under physiological-like solution conditions (1.33 mmol/liter Ca(2+), 1.0 mmol/liter total inorganic phosphate, (0 or 26) mmol/liter carbonate, 270 mmol/kg osmolality, pH 7.4, 37 degrees C). When the mass of the new accretions equaled the initial seed mass, the solids were harvested and the net change in crystal size resulting from the new accretions was assessed by X-ray diffraction line-broadening analysis. All the electrolytes examined in this study inhibited the accretion rate. The order of effectiveness was phosvitin > polyaspartate approximately polyglutamate > 1-hydroxyethylidene-1,1-bisphosphonate (HEBP) > bovine serum albumin (BSA) > citrate. Citrate and BSA also reduced the mean crystal size of the harvested solids compared with those harvested in the absence of added electrolyte, a finding that suggests that citrate and BSA suppressed growth of the seed crystals in favor of the proliferation of new smaller crystals. In contrast, a net increase in size following accretion compared with controls suggests that the other more strongly inhibiting electrolytes stimulated growth of the primary seed crystals and/or of the secondary crystals. These size changes, however, were anisotropic, with the anions effecting primarily increases in crystal width/thickness rather than in length. The effects were also more pronounced in the presence of carbonate. Our findings suggest that the strength of the interaction with the crystal surface may be relatively more important than molecular size or conformational complexity in establishing the effect that electrolytes have on apatite growth and proliferation. The results also suggest that adsorbed electrolytes may be a significant factor in controlling the size of apatite crystals in skeletal tissues by inducing proliferation of new crystals and/or affecting crystal shape by selectively modifying growth of the lateral dimensions. PMID- 10821883 TI - Changes in geometry and cortical porosity in adult, ovary-intact rabbits after 5 months treatment with LY333334 (hPTH 1-34). AB - The purpose of this study was to determine if the increased cortical bone porosity induced by intermittently administered parathyroid hormone (PTH) reduces bone strength significantly. Mature ovary-intact New Zealand white rabbits were treated with once daily injections of vehicle, or PTH(1-34), LY333334, at 10 or 40 microg/kg/day for 140 days. Geometry of the femoral midshaft was measured to evaluate changes in the cross-sectional moment of inertia (CSMI). Cortical porosity was measured in the midshaft of the tibia by dividing cortical area into three zones based on equal divisions of cortical diameter: near endocortical (Zone I), near intermediate (Zone II), and near periosteal (Zone III) regions. Total cortical porosity significantly increased after PTH treatment from 1.4% in the controls to 6.3% in the higher dose group, but the location of the new porosities was not randomly distributed. In the controls, porosity of Zones I and II (both 1.7%) was almost twice as much as that of Zone III (0.9%). In the lower dose group, cortical porosity of Zone I (5.5%) and II (1.8%) was greater than in Zone III (0.9%), but these differences were not statistically significant. In the higher dose group, cortical porosity of Zone I (11.5%) and II (6. 1%) significantly increased compared with Zone III (1.4%) (P < 0. 0005). Histomorphometric measurements showed that bone formation rate on both periosteal and endocortical surfaces increased, resulting in increased bone area and cortical area in the higher dose group. A model was developed to evaluate the effect of the changes in geometry and porosity on CSMI in the different zones. This simulation model indicated that CSMI in the higher dose group was significantly greater than in the other two groups, despite the increased porosity. We speculate the reason to be that porosity increased near the endocortical surface, where its mechanical effect is small. This increase was more than offset by apposition of new bone on the periosteal surface. These data suggest that (1) PTH increases cortical porosity in a dose-dependent manner, primarily near endocortical surfaces; (2) because of this nonhomogeneous distribution, the mechanical effect of increased porosity is small; (3) the increased cortical porosity associated with PTH treatment is more than offset by periosteal apposition of new bone, causing an overall increase in the bending rigidity of cortical bone; and (4) these changes cannot be accurately evaluated using noninvasive methods of bone densitometry, which cannot account for the location of bone gain and bone loss. PMID- 10821884 TI - The high-affinity estrogen receptor antagonist ICI 182,780 has no effect on bone growth in young male rats. AB - Estrogens have profound effects on the processes of bone formation and turnover in females. The physiological role of this class of hormones on bone metabolism in males is less certain. The purpose of this study was to determine the effect of the high affinity estrogen receptor antagonist ICI 182,780 on tibial growth in normal male rats. The effects of ICI 182,780 on growing male rats were compared to orchiectomy, which prevents the synthesis of estrogens as well as androgens. Neither orchiectomy nor ICI 182,780 had an effect on body weight gained. Orchiectomy decreased longitudinal bone growth at the proximal tibial metaphysis and radial bone growth at the tibia-fibula synostosis. In contrast, ICI 182,780 had no effect on either endochondral or intramembranous bone growth. These findings suggest that androgens are more important than estrogens in determining peak bone mass in male rats. PMID- 10821885 TI - 2-Methoxyestradiol inhibits longitudinal bone growth in normal female rats. AB - 2-Methoxyestradiol (2-MeO-E(2)), a major metabolite of 17beta-estradiol, may function as a physiological tumor suppressor and is being investigated for clinical applications. It has been reported to target rapidly dividing cells. We investigated the effects of 2-MeO-E(2) on the growth plate of young rats because normal longitudinal bone growth requires rapid proliferation of cartilage and endothelial cells. Sexually mature (3-month-old) normal female rats were treated with 2-MeO-E(2) (100 mg/kg/day) for 13 days and it was found to have no effect on uterine weight but reduced serum cholesterol. The estrogen metabolite had no effect on either cortical or cancellous bone. In contrast, 2-MeO-E(2) dramatically reduced longitudinal bone growth rate at the proximal tibia from 55 +/- 2 to 20 +/- 2 microm/day (P < 0.001) and growth plate thickness from 153 +/- 14 to 70 +/- 6 microm (P < 0.001). The latter decrease was due to significant reductions in the height of both the proliferative (P < 0.001) and the hypertrophic (P < 0.001) zones. These results in normal female rats demonstrate that 2-MeO-E(2) inhibited longitudinal bone growth but had no effect on either radial bone growth or cancellous bone turnover. 2-MeO-E(2) was shown by these studies to have the ability to discriminate between bone and cartilage, as well as between reproductive and nonreproductive estrogen-target tissues. Thus, 2-MeO E(2) is a naturally produced estrogen metabolite that demonstrates unique tissue selectivity. PMID- 10821886 TI - Gonadal steroids and bone metabolism in young castrated male rats. AB - At 45 days of age, 40 male Wistar rats were castrated, then randomly divided into four groups, S.C. injected for 60 days after surgery either with 17beta-estradiol (E) 10 microg/kg BW/48 hours, progesterone (P) 140 microg/kg BW/48 hours, dihydrotestosterone (D) 2 microg/kg BW/48 hours, E + P + D same doses, or solvent alone (CX). Ten other rats were sham-operated (SH) and used as controls. Animals were put in balance to determine Ca and phosphorus (Pi) intestinal apparent absorption (IA Ca, IA Pi) and urinary pyridinium crosslinks excretion. Plasma was collected for measurement of intact-parathyroid hormone (PTH), calcitonin (CT), insulin-like growth factor I (IGF-I), 1,25 dihydroxyvitamin D (1,25(OH)(2)D), Ca, and Pi. Orchidectomy induced marked seminal vesicles atrophy and increased plasma CT, PTH, and Ca concentrations. IA Ca was significantly higher in P rats, however, neither castration nor any other treatment had significant effects. Orchidectomy decreased femoral length, dry weight, and Ca content, whereas E or D given alone or together with P improved endochondral growth and enhanced femoral Ca content. Again, bone mineral density was lowered by orchidectomy and reestablished by both E and EPD, even above SH values, this effect being more important at the metaphyseal levels. Urinary pyridinium cross-links excretion and plasma osteocalcin concentrations were higher in the CX animals than in the controls. Although E and D given alone did reduce both biochemical turnover markers, they showed additive effect when given together (EPD). In conclusion, in the young castrated male rat, E was more efficient than D for preventing bone loss, the most important effect being induced by a combination of E+P+D. PMID- 10821888 TI - European calcified tissue society PMID- 10821887 TI - Vitamin D deficiency and susceptibility to tuberculosis. AB - Vitamin D, a modulator of macrophage function, can activate human anti mycobacterial activity. Vitamin D deficiency is therefore associated with a higher risk of tuberculosis (TB) infection, as indicated by several observations. First, TB tends to occur during the colder seasons when cutaneous synthesis of vitamin D from sun exposure is reduced and serum vitamin D levels are lower. Second, patients with untreated TB, particularly those from a temperate climate, have lower serum vitamin D levels than healthy subjects. Third, the incidence of TB is higher among subjects with relatively low serum vitamin D levels, such as the elderly, uremic patients, and Asian immigrants in the U.K. PMID- 10821889 TI - Fluoroscopically guided transcervical fallopian tube recanalization of post sterilization reversal mid-tubal obstructions. AB - PURPOSE: To assess the technical success and early outcome of fluoroscopically guided transcervical fallopian tube recanalization (FTR) in mid-tubal occlusion following sterilization reversal surgery. METHODS: From July 1995 to January 1998, patients with greater than 12 months secondary infertility underwent hysterosalpingography (HSG). FTR was performed in proximal or mid-tubal occlusion. Cases of FTR in mid-tubal occlusion were included in this study. Technical success (defined as complete tubal patency) using a standard guidewire and hydrophilic glidewire, the number of patients with at least one patent tube, and the intrauterine and ectopic pregnancy rates were determined. RESULTS: Twenty six infertile patients with previous sterilization reversal underwent HSG. Eight of 26 (31%) patients (mean age 32 years, range 23-37 years), had attempted FTR for mid-tubal occlusion at the site of surgical anastomosis. Fourteen tubes were attempted as there were two previous salpingectomies. Technical success was achieved in eight of 14 (57%) tubes attempted, resulting in five of eight (62%) patients having at least one patent tube. At follow-up (mean 18 months, range 12 28 months) in these five patients there was one intrauterine pregnancy. There were no ectopic pregnancies. CONCLUSIONS: FTR in mid-tubal obstruction in infertile patients following sterilization reversal surgery is technically feasible and may result in intrauterine pregnancy. In this small group there was a lower technical success rate and lower pregnancy rate than in unselected proximal tubal occlusion. PMID- 10821890 TI - Acetic acid as a sclerosing agent for renal cysts: comparison with ethanol in follow-up results. AB - PURPOSE: To compare follow-up results of sclerotherapy for renal cyst using 50% acetic acid with those using 99% ethanol as sclerosing agents. METHODS: Eighty one patients underwent sclerotherapy and 58 patients, 23 males, 35 females, aged 6-76 years, having a total of 60 cysts, were included in this study; the others were lost to follow-up. The renal cysts were diagnosed by sonography, computed tomography (CT), or magnetic resonance imaging (MRI). Sclerotherapy was performed using 50% acetic acid for 32 cysts in 31 patients and 99% ethanol for 28 cysts in 27 patients. Under fluoroscopic guidance, cystic fluid was aspirated as completely as possible. After instillation of a sclerosing agent corresponding to 11.7%-25% (4-100 ml) of the aspirated volume, the patient changed position for 20 min and then the agent was removed. Patients were followed up by sonography for a period of 1-49 months. The volume of the renal cyst after sclerotherapy was compared with that of the renal cyst calculated before sclerotherapy. Medical records were reviewed to analyze complications. RESULTS: The mean volume after sclerotherapy of the 17 cysts followed for 3-4 months in the acetic acid group was 5.1% of the initial volume, and for the 14 cysts in the ethanol group it was 10.2%. Complete regression during follow-up was shown in 21 cysts (66%) in the acetic acid group; the mean volume of these cysts before the procedure was 245 ml. The mean volume of the nine (32%) completely regressed cysts in the ethanol group was 184 ml. Mild flank pain, which occurred in three patients in each group, was the only complication and resolved the next day. CONCLUSION: Acetic acid was an effective and safe sclerosing agent for renal cysts, tending to induce faster and more complete regression than ethanol. PMID- 10821891 TI - Instability of reference diameter in the evaluation of stenosis after coronary angioplasty: percent diameter stenosis overestimates dilative effects due to reference diameter reduction. AB - PURPOSE: To examine changes in the reference segment luminal diameter after coronary angioplasty. METHODS: Sixty-one patients with stable angina pectoris or old myocardial infarction were examined. Coronary angiograms were recorded before coronary angioplasty (pre-angioplasty) and immediately after (post-angioplasty), as well as 3 months after. Artery diameters were measured on cine-film using quantitative coronary angiographic analysis. RESULTS: The diameters of the proximal segment not involved in the balloon inflation and segments in the other artery did not change significantly after angioplasty, but the reference segment diameter significantly decreased (4.7%). More than 10% luminal reduction was observed in seven patients (11%) and more than 5% reduction was observed in 25 patients (41%). More than 5% underestimation of the stenosis was observed in 22 patients (36%) when the post-angioplasty reference diameter was used as the reference diameter, compared with when the pre-angioplasty measurement was used and more than 10% underestimation was observed in five patients (8%). CONCLUSION: This study indicated that evaluation by percent diameter stenosis, with the reference diameter from immediately after angioplasty, overestimates the dilative effects of coronary angioplasty, and that it is thus better to evaluate the efficacy of angioplasty using the absolute diameter in addition to percent luminal stenosis. PMID- 10821892 TI - Peripheral venous access ports: outcomes analysis in 109 patients. AB - PURPOSE: To perform a retrospective outcomes analysis of central venous catheters with peripheral venous access ports, with comparison to published data. METHODS: One hundred and twelve central venous catheters with peripherally placed access ports were placed under sonographic guidance in 109 patients over a 4-year period. Ports were placed for the administration of chemotherapy, hyperalimentation, long-term antibiotic therapy, gamma-globulin therapy, and frequent blood sampling. A vein in the upper arm was accessed in each case and the catheter was passed to the superior vena cava or right atrium. Povidone iodine skin preparation was used in the first 65 port insertions. A combination of Iodophor solution and povidone iodine solution was used in the last 47 port insertions. Forty patients received low-dose (1 mg) warfarin sodium beginning the day after port insertion. Three patients received higher doses of warfarin sodium for preexistent venous thrombosis. Catheter performance and complications were assessed and compared with published data. RESULTS: Access into the basilic or brachial veins was obtained in all cases. Ports remained functional for a total of 28,936 patient days. The port functioned in 50% of patients until completion of therapy, or the patient's expiration. Ports were removed prior to completion of therapy in 18% of patients. Eleven patients (9.9% of ports placed) suffered an infectious complication (0.38 per thousand catheter-days)-in nine, at the port implantation site, in two along the catheter. In all 11 instances the port was removed. Port pocket infection in the early postoperative period occurred in three patients (4.7%) receiving a Betadine prep vs two patients (4.2%) receiving a standard O.R. prep. This difference was not statistically significant (p = 0.9). Venous thrombosis occurred in three patients (6.8%) receiving warfarin sodium and in two patients (3%) not receiving warfarin sodium. This difference was not statistically significant (p = 0.6). Aspiration occlusion occurred in 13 patients (11.7%). Intracatheter urokinase was infused in eight of these patients and successfully restored catheter function in all but two instances. These complication rates are comparable to or better than those reported with chest ports. CONCLUSION: Peripheral ports for long-term central venous access placed by interventional radiologists in the interventional radiology suite are as safe and as effective as chest ports. PMID- 10821893 TI - Thrombus on indwelling central venous catheters: the histopathology of "Fibrin sheaths". AB - PURPOSE: Central venous catheters (CVC) may fail for many reasons, though "fibrin sheaths" blocking catheter ports are usually implicated. We examined the sheaths removed from dialysis catheters to determine their histopathology. METHODS: Ten catheter strippings were performed and the removed material was studied grossly and microscopically. RESULTS: The histologic specimens showed thrombus both with and without a proteinaceous sheath. CONCLUSION: Dialysis catheters fail because of thrombus formation. This can occur in either the absence or presence of a protein coating on the catheter, the so-called "fibrin sheath." PMID- 10821894 TI - Subtraction CT with low-flow-rate arterial contrast injection to estimate drug distribution during balloon-occluded arterial chemotherapy infusion for bladder cancer. AB - PURPOSE: To simulate drug distribution during balloon-occluded arterial chemotherapy infusion (BOAI) for urinary bladder cancer using subtraction computed tomography (CT) with low-flow-rate arterial contrast injection (S-CTLA). METHODS: Ten patients with bladder cancer underwent S-CTLA, and the distribution of contrast agent during BOAI into both internal iliac arteries simultaneously was evaluated in nine pairs of internal iliac arteries and one single artery. For S-CTLA, spiral CT data were acquired before and after 0.2 ml/sec intraarterial injection of contrast material. The enhancement of the urinary bladder wall, the gluteal muscles, and the pelvic bones was categorized using a 4-grade scale. The grades were compared in each of the three pelvic components and differences were tested for significance using the Wilcoxon test for paired groups. RESULTS: S CTLA revealed the distribution of the contrast agent clearly. Gluteal muscles grades were significantly higher than those of the other two assessed components. CONCLUSION: BOAI does not improve the concentration of contrast agent to the bladder wall over neighboring structures, suggesting that the balloon occlusion technique does not achieve its desired goal for chemotherapy targeting. PMID- 10821895 TI - Percutaneous implantation of a catheter with subcutaneous reservoir for intraarterial regional chemotherapy: technique and preliminary results. AB - PURPOSE: We present the technique and the preliminary results of percutaneous implantation of intraarterial catheters connected to a subcutaneous infusion reservoir for prolonged regional chemotherapy of hepatic and extrahepatic tumors. METHODS: Two hundred patients with primary or secondary hepatic neoplasms, pelvic, pancreatic, renal, lingual, and breast cancer underwent the procedure. The access was the left axillary artery (188 patients) and the femoral artery (12 patients). The catheter tip was placed in the hepatic (170 patients), hypogastric (18), splenic (4), internal thoracic (2), gastroduodenal (3), renal (2) or the external carotid artery (1). The catheter was connected to a subcutaneous reservoir and filled with heparin; chemotherapeutic infusion was subsequently started. RESULTS: One hundred percent immediate technical success was obtained. Forty-three of 200 (21.5%) patients had a complication: 29 patients had a catheter dislodgment, nine had arterial thrombosis, three had a pseudoaneurysm of the left axillary artery and two had a port pocket hematoma. Most complications (37/43, 86%) were treated percutaneously without interruption of chemotherapy. In only six cases (3% of the total population) was chemotherapy discontinued due to the complication itself. The mean duration of catheter patency was 7.2 months. CONCLUSION: Percutaneous placement of an intraarterial catheter is feasible and causes less discomfort to the patient than the surgical approach. The technique has an acceptable complication rate (21.5%), similar to that for surgical implantation (17.8%), with the advantage that in most cases the complications can be resolved percutaneously. This technique represents an alternative to surgical implantation in the treatment of liver metastases from colorectal cancer and opens new therapeutic possibilities for the local prolonged treatment of other kinds of tumor, though its clinical efficacy must be assessed in selected trials. PMID- 10821896 TI - Growth characteristics and imaging properties of the morris hepatoma 3924A in ACI rats: a suitable model for transarterial chemoembolization. AB - PURPOSE: For experimental studies investigating modalities and efficacy of transarterial chemoembolization (TACE) in hepatocellular carcinoma (HCC) an animal model resembling the human situation as closely as possible would be appropriate. Specifically, reproducible tumor growth characteristics with the capability for appropriate in vivo imaging to monitor treatment efficacy are required. METHODS: Morris hepatoma 3924A was implanted into the liver of 30 ACI rats. Tumor growth was followed by angiography (n = 10), ultrasound (US, n = 30), native computed tomography (CT, n = 16), and native magnetic resonance imaging (MRI, n = 30) between day 8 and day 36 after implantation. The radiological morphological characteristics were compared with the macroscopic and microscopic histological findings of the explanted tumors. RESULTS: In all 30 animals a solitary liver tumor was found and macroscopically no signs of metastases, ascites, or peritoneal tumor were visible. On histopathological examination tumor sizes ranged between 27 +/- 3 mm(3) (day 8) and 3468 +/- 79 mm(3) (day 36). The first signs of tumor necrosis occurred at day 16. US allowed tumor visualization from day 8, MRI from day 8, angiography from day 10, and CT from day 14. CONCLUSIONS: The tumor model has the potential to be used for the visualization of tumor growth by MRI and US. The potential for monitoring therapeutic effects of TACE needs to be investigated. PMID- 10821897 TI - Experimental embolization of rabbit renal arteries to compare the effects of poly L-lactic acid microspheres with and without epirubicin release against intraarterial injection of epirubicin. AB - PURPOSE: We performed a basic investigation using white rabbits of the sustained release and embolizing effects of poly L-lactic acid microspheres (PLA) to determine their usefulness for chemoembolization. METHODS: Fifteen male Japanese white rabbits were used. Sustained release of an embolizing material, EPI-PLA was accomplished with 1 mg of PLA containing 0.03 mg of epirubicin hydrochloride (EPI). Embolization with 50 mg of PLA (total dose of EPI 1.5 mg) was performed after the renal artery of the rabbits was selected (Chemo-TAE group). A group in which a bolus of 1.5 mg EPI alone was injected through the renal artery (TAI group) was established as a control group. Furthermore, a group in which embolization was performed with 50 mg of PLA alone (TAE group) was also established. These three groups, each consisting of five rabbits, were compared. RESULTS: Blood EPI levels were serially measured. The blood EPI level in the TAI group rapidly reached a peak more than 30 min after injection, then decreased to almost zero 24 hr after injection. In the Chemo-TAE group, the blood EPI level was transiently increased 30 min after embolization, but remained low thereafter until 24 hr after embolization. EPI levels in kidney tissue isolated 24 hr after embolization were measured. In the Chemo-TAE group, the tissue EPI level was significantly higher than that in the TAI group. When isolated kidneys were macroscopically and histologically examined, atrophy of the entire embolized kidney, as well as infarction and necrosis in the renal cortex, were observed in both the TAE group and the Chemo-TAE group. However, there were no such findings in the TAI group. The area of the infarction in the renal cortex did not significantly differ between the Chemo-TAE group and the TAE group; however, there was vascular injury in the Chemo-TAE group and none in the TAE group. CONCLUSION: It was demonstrated that EPI-PLA, a chemoembolizing material, maintained high local concentrations of the anticancer drug, while maintaining low blood levels of the anticancer drug. PMID- 10821898 TI - Percutaneous untying of a knot in a retained Swan-Ganz catheter. AB - A patient was referred to us with a tightly knotted Swan-Ganz catheter. The catheter could not be removed by conventional simple methods. We describe a minimally invasive means of removal of the catheter using an Amplatz gooseneck snare and an angioplasty balloon. This allowed the catheter to be removed without trauma. PMID- 10821899 TI - Relief of buttock claudication by percutaneous recanalization of an occluded superior gluteal artery. AB - We report a case of a woman presenting with right severe buttock claudication and normal neurological and osteoarticular examination, in whom a guidewire recanalization and percutaneous transluminal angioplasty (PTA) of an occluded right superior gluteal artery (SGA) has provided relief of her symptoms. To our knowledge, this is the first report of percutaneous recanalization of the SGA. PTA can be considered the treatment of choice for buttock claudication caused by SGA stenosis or occlusion. PMID- 10821900 TI - Balloon-assisted coil embolization for large-necked renal artery aneurysms. AB - An aneurysm of the right renal artery was discovered in a patient suffering from cerebral arterial angiodysplasia and arterial hypertension. The aneurysm was large necked, which made selective endovascular treatment very difficult. To perform the embolization of the aneurysm, a balloon remodelling technique was used. This prevented migration of coils within the arterial lumen. PMID- 10821901 TI - Endovascular treatment of a portal vein tear during TIPSS. AB - During a transjugular portosystemic stent-shunt (TIPSS) procedure a portal vein laceration occurred with subsequent intraperitoneal hemorrhage. A PTFE-covered nitinol stent was successfully placed eliminating the leak and creating a functioning portosystemic shunt. This case demonstrates both the importance of portal vein puncture more than 1 cm from the bifurcation and the necessity of maintaining a stock of available stent-grafts. PMID- 10821902 TI - Transjugular intrahepatic portosystemic shunt for the treatment of intractable ascites in a patient with polycystic liver disease. AB - Though polycystic liver disease (PCLD) has historically been considered a contraindication to TIPS, we present a case where technically successful shunt creation was achieved without the need for modification of the standard TIPS procedure, as was required in a previous report. PMID- 10821903 TI - PMMA cementoplasty in symptomatic metastatic lesions of the S1 vertebral body. AB - We describe a lateral transiliac direct puncture approach to the S1 vertebral body for polymethylmethacrylate (PMMA) cementoplasty of painful metastatic lesions. This approach was performed using a 15-cm-long trocar needle with 3-mm outer diameter, introduced under general anesthesia and fluoroscopic control. A lateral projection was used to center the needle just in front of the spinal canal and subjacent to the superior plate of the S1 vertebral body. Needle progression was controlled using anteroposterior and lateral fluoroscopic projections alternately with a needle course parallel to an axial plane, avoiding conflict with the S1 foramen. After needle tip placement in the center of the S1 vertebral body, diluted PMMA with a setting time of 8 min was delivered. Ipsilateral lesions of the lateral sacral compartment were filled with the same needle by stepwise withdrawal and continuous PMMA injection. PMID- 10821904 TI - Use of through-and-through guidewire for delivering large stent-grafts into the distal aortic arch. AB - The availability of large diameter stent-grafts is now allowing the endovascular treatment of thoracic aortic aneurysms. Most aneurysms are closely related to the distal arch and it is thus necessary to pass the delivery systems into the arch to effectively cover the proximal neck. Even with extra-stiff guidewires in position, it may still be difficult to achieve this, as a result of tortuosity at the iliac arteries and the aorta. We detail a technique where a stiff guidewire is passed from a brachial entry point through the aorta and out at the femoral arteriotomy site. This allows extra-support and may enable the delivery system to be passed further into the aortic arch than it could with just the regular guidewire position. PMID- 10821905 TI - Endoluminal embolization of bilateral atherosclerotic common iliac aneurysms with fibrin tissue glue (Beriplast). AB - The standard surgical approach to nonleaking iliac aneurysms found at repair of a leaking abdominal aortic aneurysm is to minimize the operative risk by repairing the abdominal aorta only. This means that the bypassed iliac aneurysms may have to be repaired later. As this population of patients are usually elderly with coexisting medical problems, interventional radiology is being used to embolize these aneurysms, thus avoiding the morbidity and mortality associated with further general anesthesia and surgery. Various materials and stents have been reported to be effective in the treatment of iliac aneurysms. We report the successful use of endoluminal fibrin tissue glue (Beriplast) to treat two large iliac aneurysms in a patient who had had a previous abdominal aortic aneurysm repair. We discuss the technique involved and the reasons why we used tissue glue in this patient. PMID- 10821906 TI - Stent placement for malignant pulmonary artery structure. PMID- 10821916 TI - Reduction of infarct size by the NO donors sodium nitroprusside and spermine/NO after transient focal cerebral ischemia in rats. AB - Nitric oxide (NO) plays a dual role (neuroprotection and neurotoxicity) in cerebral ischemia. NO promoting strategies may be beneficial shortly after ischemia. Therefore, we have studied the hemodynamic and possible neuroprotective effects of two NO donors, the classical nitrovasodilator sodium nitroprusside (SNP) and the NONOate spermine/NO, after transient focal cerebral ischemia in rats. Parietal cortical perfusion was measured by laser-Doppler flowmetry. The effects of increasing intravenous doses (10-300 microgram) of sodium nitroprusside and spermine/NO on cortical perfusion and arterial blood pressure were assessed. Transient (2 h) focal cerebral ischemia was carried out by the intraluminal thread method. The effects of intraischemic intravenous infusion of SNP (0.11, 1.1 mg/kg) and spermine/NO (0.36, 3.6 mg/kg) on hemodynamic parameters and infarct size developed after 1 week reperfusion were assessed. In control conditions, SNP and, to a lesser extent, spermine/NO induced dose-dependent hypotension and concomitant reduction in cortical perfusion. In focal cerebral ischemia, infusion of SNP (0.11 mg/kg) and spermine/NO (0.36, 3.6 mg/kg) reduced the infarct size. In the case of spermine/NO, cortical perfusion was maintained above the control levels during the ischemic insult. No significant hypotension was elicited by NO donors at the dose-ratios infused. In conclusion, brain damage induced by transient focal ischemia is reduced by intravenous NO donors. Neuroprotective effects of spermine/NO are due at least in part to improvement of brain perfusion, while sodium nitroprusside must provide direct cytoprotection. These results give further support to the protective effect of NO in the early stages of cerebral ischemia and point to the therapeutic potential of NONOates in the management of brain ischemic damage. PMID- 10821917 TI - Amyloid precursor protein and membrane phospholipids in primary cortical neurons increase with development, or after exposure to nerve growth factor or Abeta(1 40). AB - We examined the relationships between membrane phospholipid levels, the secretion and expression of the amyloid precursor protein (APP), and the responses of both to nerve growth factor (NGF), Abeta(1-40) or Abeta(40-1) in developing cortical neurons cultured from rat embryos. Neuronal membrane phospholipid levels per cell, and phosphatidylcholine, phosphatidylserine, phosphatidylinositol and phosphatidylethanolamine increased individually between the first and seventh days of culturing. The amounts of APP holoprotein and APP mRNAs in the cells, as well as the amounts of soluble APP (APPs) secreted by them, also increased during neuronal development in vitro. The increases in APPs exceeded the increases in APP which, in turn, exceed those in phospholipid levels. The levels of APP holoprotein, but not of phospholipids, increased when neurons were grown in a choline-free medium, suggesting that increases in APP are not sufficient to stimulate changes in membrane phospholipids. Treatment of neuron cultures for four days with NGF or Abeta(1-40), but not with Abeta(40-1), dose-dependently increased membrane phospholipids, tau and GAP-43, as well as APP holoprotein and secreted APPs. These results indicate that agents, like NGF or Abeta(1-40), which enhance membrane phospholipid levels may promote neurite formation, APP expression and APPs secretion in primary neuronal cultures. PMID- 10821918 TI - Galanin-immunoreactive neuronal system and colocalization with serotonin in the optic lobe and peduncle complex of the octopus (Octopus vulgaris). AB - Immunohistochemical techniques were used to investigate the distribution of galanin-like immunoreactivity and colocalization with serotonin (5-HT) in the optic lobe and peduncle complex of the octopus, Octopus vulgaris. Galanin immunoreactive (Gal-IR) fibers, but not cells, were seen in the plexiform layer of the optic lobe cortex. Gal-IR cells were scattered in the cell-islands of the optic lobe medulla and Gal-IR varicose fibers were observed to be abundant in the neuropil surrounding the islands. All Gal-IR cells were immunoreactive for 5-HT, and a few cells showed only 5-HT-like immunoreactivity. In the peduncle lobe, no Gal-IR cells were seen in the basal zone or spine, but in the basal zone, many Gal-IR fibers were seen. In the anterior olfactory lobule, only a few pyramidal Gal-IR cells were observed in the cell layer, and their apical processes were traced to the central neuropil. In the median olfactory lobule, ovoid Gal-IR cells were scattered in the peripheral cell layer. All Gal-IR cells in the anterior and median olfactory lobules showed 5-HT-like immunoreactivity. In the posterior olfactory lobule, ovoid and triangular Gal-IR cells were scattered in the cell layer. Some of them showed 5-HT-like immunoreactivity. Western blot analysis indicated an Gal-IR band at approximately 15.4 kDa. These results suggest the association of galanin-like substance and 5-HT with the visual system of octopus and that the main form of the octopus galanin might have a different molecular weight from vertebrate galanins. PMID- 10821919 TI - Rostral ventrolateral medulla opioid receptor activation modulates glutamate release and attenuates the exercise pressor reflex. AB - We previously reported that the administration of [D-Ala(2)]methionine enkephalinamide (DAME), an opioid receptor agonist, into the rostral (RVLM) but not into the caudal ventrolateral medulla (CVLM), attenuated increases in mean arterial pressure (MAP) and heart rate (HR) during static muscle contraction that had been blocked by prior microdialysis of the opioid receptor antagonist, naloxone [Am. J. Physiol. 274 (1998) H139-H146]. In this study, we determine whether this RVLM-mediated opioidergic-modulation of cardiovascular responses is associated with localized changes in extracellular concentrations of glutamate, an excitatory amino acid, using microdialysis techniques in anesthetized rats. Muscle contraction increased MAP and HR by 37+/-5 mmHg and 23+/-3 bpm, respectively. Extracellular glutamate concentrations, determined using HPLC-ECD, increased from 0.8+/-0.2 to 6.6+/-1.2 ng/5 microliter in the bilateral RVLM areas. Microdialysis of DAME (100 microM) for 30 min attenuated the contraction evoked increases in MAP, HR, and glutamate levels (20+/-4 mmHg, 10+/-2 bpm, and 1.8+/-0.2 ng/5 microliter, respectively). After microdialysis of naloxone (100 microM) for 30 min into the RVLM, muscle contraction blocked the attenuations (35+/-5 mmHg, 26+/-4 bpm, and 5.8+/-1.0 ng/5 microliter, respectively). Developed muscle tensions were similar throughout the protocol (676+/-38, 678+/-37 and 687+/-37 g, respectively). These results suggest that an opioidergic receptor mediated mechanism within the RVLM attenuates cardiovascular responses during static exercise via modulating extracellular concentrations of glutamate in the RVLM. PMID- 10821920 TI - Involvement of GABA(A) and GABA(B) receptors in afterdischarge generation in rat hippocampal slices. AB - It has been hypothesized that a disruption of gamma-aminobutyric acid (GABA) receptor-mediated processes may be involved in the pathophysiology of focal epilepsy. This disinhibition hypothesis has been postulated from the results of in vitro experiments of the interictal activity of focal epilepsy. Less is known, however, about how disinhibition may be involved in the production of the ictal activity. We therefore examined the pharmacological effects of selective agonists and antagonists of GABA(A) and GABA(B) receptors on ictal-like afterdischarges (ADs) induced following repetitive high-frequency electrical stimulation in the CA1 region of rat hippocampal slices. The GABA(A) receptor antagonist bicuculline (5 microM) fully blocked AD generation, as did the GABA(A) receptor agonist muscimol (2 microM), which is thought to produce a tonic inhibition during application. However, the benzodiazepine receptor agonist diazepam (5 microM), which enhances the inhibitory postsynaptic potential induced by synaptically released GABA, increased the number of spikes in the AD to 148.3% of the control value. On the other hand, the GABA(B) receptor antagonist phaclofen (1 mM) increased the number of spikes in the AD to 234.7% of the control value, while the GABA(B) receptor agonist baclofen (5 microM) reduced it to 46.9%. We therefore conclude that synaptic, but not tonic, activation of GABA(A) receptors appears to be necessary for ictal-like AD generation, while GABA(B) receptor activation plays a protective role. We therefore propose a modification to the simple disinhibition hypothesis. PMID- 10821921 TI - Cue-induced changes in basal local cerebral glucose utilization 13 days after morphine sensitization in the Fischer 344 rat: relevance for drug craving. AB - The present experiment tested the hypothesis that some persistent neural adaptation develops during the course of repeated sensitizing doses of morphine administered to rats. A sub-hypothesis was that this imprint would be of greater magnitude in the presence of morphine-conditioned cues. In order to test these hypotheses basal local cerebral metabolic rates for glucose (LCMR(glu)) were determined 13 days after the last of four 10-mg/kg doses of morphine administered in 36 h to Fischer 344 male rats. LCMR(glu) was determined using the 2-deoxy-D-[1 (14)C]glucose method (2-DG). Half of the rats, the conditioned group, were placed in the 2-DG chamber after each injection and half, the nonconditioned group, were placed in a neutral environment. A control group received only saline in lieu of morphine. All metabolic rates were determined in a nondrugged state. The major finding was large increases in metabolic rate throughout the forebrain in the sensitized rats. This was especially so in the conditioned group, 46 out of 93 areas examined had significant increases while in the nonconditioned group it was 25 out of 93. Both the core and shell of the nucleus accumbens showed significant elevations in metabolic rates in the presence of morphine cues but only the shell in the absence of the cues. There were no significant decreases in basal metabolic activity in any of the brain regions evaluated in either experimental group. The present finding suggests that changes in the brains of these morphine sensitized rats may model the altered brain states responsible for drug craving in human drug addicts. PMID- 10821922 TI - Lateral asymmetries in the trigeminal ganglion of the male rat. AB - We have applied stereological methods to estimate the number and perikaryal size of primary sensory neurons in celloidin-embedded trigeminal ganglia of male albino rats, specifically looking for inter-individual and side variability. The mean total number of neurons per ganglion was 35,300, with a moderate variability among ganglia. On average, 66% of the neurons were classified as A-type and 34% as B-type. Although for individual cases there could be notable side differences in the number of neurons of each type, on a population basis these differences were not significant. Mean neuronal volume was four times larger for A- than for B-cells, and both populations exhibited a moderate variability among individuals. High intra-animal side differences were found for A-cells, which were on average a significant 23.5% larger in the right ganglia. B-cells did not show significant side differences. The distribution of individual volumes around the mean value was consistently skewed to the right, particularly in the case of A-cells, which partially overlapped with the largest B-cells. In the right ganglion the distribution of A-cells, but not of B-cells, showed a rightward bias, revealing the increase in bigger neurons. The existence of larger A-type neurons in the right trigeminal ganglion may provide a structural substrate for some somesthetically based complex behaviors which are best performed by rats using their right vibrissae. PMID- 10821923 TI - Effects of nitric oxide donors on the afferent resting activity in the cephalopod statocyst. AB - The effects of bath applications of the nitric oxide (NO) donors sodium nitroprusside (SNP), diethylamine sodium (DEA), 3-morpholinosydnonimine (SIN-1), and S-nitroso-N-acetyl-penicillamine (SNAP) on the resting activity (RA) of afferent crista fibers were studied in isolated statocysts of the cuttlefish Sepia officinalis. The NO donors had three different effects: inhibition, excitation, and excitation followed by an inhibition. The SNAP analog N-acetyl-DL penicillamine (xSNAP; with no NO moiety) had no effect. When the preparation was pre-treated with the NO synthase inhibitor N(G)-nitric-L-arginine methyl ester HCl (L-NAME), the NO donors were still effective. When the preparation was pre treated with the guanylate cyclase inhibitors methylene blue (M-BLU) or cystamine (CYS), NO donors had only excitatory effects, whereas their effects were inhibitory only when pre-treatment was with the adenylate cyclase inhibitors nicotinic acid (NIC-A), 2',3'-dideoxyadenosine (DDA), or MDL-12330A. When pre treatment was with a guanylate and an adenylate cyclase inhibitor combined, NO donors had no effect; in that situation, the RA of the afferent fibers remained and the preparation still responded to bath applications of GABA. Selective experiments with statocysts from the squid Sepioteuthis lessoniana and the octopod Octopus vulgaris gave comparable results. These data indicate that in cephalopod statocysts an inhibitory NO-cGMP and an excitatory NO-cAMP signal transduction pathway exist, that these two pathways are the key pathways for the action of NO, and that they have only modulatory effects on, and are not essential for the generation of, the RA. PMID- 10821924 TI - Asymmetry of the human visual field in magnetic response to apparent motion. AB - Predominance of the lower visual field has been shown in various visual tasks, but whether the upper visual field is involved in a specific neural process is unknown. We used magnetoencephalography to study the effect of orientation and direction on the responses of five subjects to apparent motion from the human extrastriate cortex. The first magnetic response always was the largest, and the peak latency of about 200 ms did not change with the stimulus conditions. Amplitudes of the first responses were highest when motions were oriented at the horizontal meridian, decreasing with the degree of the angle between motion orientation and the horizontal meridian. There was no difference in amplitude between the two directions in the lower visual field, whereas the value of the response to downward motion in the upper visual field was significantly larger than that to upward motion. These amplitude changes are not due to differences in the anatomical distribution of neural activities because the estimated origins for the first responses always were in the same cortical area (around the occipito-parieto-temporal region) and the directions of the current vectors did not change with the stimulus conditions, and the estimated current strength changed with the stimulus conditions as did the response amplitude. These findings suggest that the human extrastriate cortex has a directional preference for downward versus upward motion in the upper visual field. PMID- 10821925 TI - Zinc-enriched (ZEN) terminals in mouse olfactory bulb. AB - The present study was designed to localize zinc-enriched (ZEN) terminals in mouse olfactory bulb by means of ZnT3 immunocytochemistry (ICC) and zinc autometallography (AMG). The immunocytochemical staining of ZnT3 was closely correlated with the AMG pattern. ZEN terminals were defined as terminals showing both ZnT3 immunoreactivities and AMG granules. At the light microscopic level, dense staining patterns for ZnT3 immunoreactivity were seen in the granule cell layer and the olfactory glomerular layer. At the ultrastructural level, ZEN terminals were restricted to presynaptic terminals with single or multiple postsynaptic thickenings. The postsynaptic profiles contacting ZEN terminals appeared to be dendrites or somata of granule cells in the granule cell layer and periglomerular cells and mitral/tufted (M/T) cells in the olfactory glomerular layer. This suggests that two main sources of ZEN terminals are present in mouse olfactory bulb: (1) centrifugal fibres making asymmetrical synapses with granule cells and periglomerular cells, and (2) olfactory receptor terminals contacting dendritic profiles of M/T cells or periglomerular cells. The close correlation between ZEN terminals and the glutamatergic system is discussed. PMID- 10821926 TI - Quantitative assessment of the normal cerebral microvasculature by endothelial barrier antigen (EBA) immunohistochemistry: application to focal cerebral ischemia. AB - Cerebrovascular endothelium participates importantly in the pathophysiology of ischemic injury. Endothelial barrier antigen (EBA) is a protein located in the luminal plasma membrane of normal central and peripheral nervous-system endothelium. In this study, we assessed the sensitivity and specificity of EBA as a quantitative marker of normal endothelium and characterized alterations of EBA immunohistochemistry following focal cerebral ischemia. Anesesthetized, non ischemic control rats (N=6) were studied. Other animals (N=5) received 90 min of middle cerebral artery occlusion (MCAo) followed by 3-day survival. Brains were prepared by perfusion-fixation and paraffin-embedding. For EBA immunohistochemistry, a monoclonal antibody (1:2000 dilution) was used. Adjacent sections were reacted for activated microglia by isolectin immunochemistry. Morphometric image-analysis was carried out in standardized microscopic fields. In control brains, pial and parenchymal blood vessels of all sizes were distinctly and selectively immunolabeled for EBA; background staining was absent. EBA-positive vascular profiles occupied 4.3+/-0.36% (mean+/-S.D.) of the microscopic field. The mean area of each identified profile was 51+/-13 micromter(2). The low coefficients of variation for both numbers of profiles (17%) and fractional areas (8%) denoted high inter-animal consistency. In brains with prior MCAo, numbers of EBA-immunoreactive vascular profiles in infarcted cortex and striatum were reduced by 39 and 46%, respectively, and their fractional areas were decreased by 63 and 76%, respectively, compared to contralateral hemisphere. Activated microglia were prominent in zones of frank infarction and in adjacent paramedian cortex; the latter region, however, showed normal-appearing EBA-immunostaining. EBA-immunohistochemistry provides a sensitive and specific index of normal cerebrovascular endothelial structures of all sizes. The technique lends itself well to quantitative morphometry and is applicable to perfusion-fixed paraffin-embedded material. EBA immunoreactivity declines in zones of ischemic infarction. PMID- 10821927 TI - Long-term evolution of local, proximal and remote astrocyte responses after diverse nucleus basalis lesioning (an experimental Alzheimer model): GFAP immunocytochemical study. AB - A study on long-term astrocytic responses (from 1 day to 20 months after lesioning in 4-month-old rats, and from 1 day to 6 months in 20-month-old rats) to diverse unilateral damage of the nucleus basalis (nbM) by injection of 40 nmol of ibotenic acid, or 50 or 100 nmols of quisqualic acid was performed using a histochemical method (immunoreactivity against the glial fibrillary acidic protein GFAP). Glial reactivity (i.e., isolated or clustered hypertrophic and/or hyper-reactive astrocytes) was evaluated in several ipsilateral and contralateral brain regions: the 'local response' within the damaged nbM region; the 'proximal response' (a new concept proposed by us) in the non-damaged structures neighbouring the nbM; and the 'remote response' in the ipsilateral brain cortex and in the contralateral cortex and nbM. In 4-month-old animals, the remote cortical glial responses, independent of the involution of cortical cholinergic activity and randomly located in layers I-V of motor and somatosensory cortical regions, were similar in appearance over a long period (13-20 months), with the highest reactivity 45 days after lesioning. The proximal response lasted from 1 day to 13 months and afterwards tended to disappear. Contralateral reactivity and ipsilateral cortical scars were observed. The local (nbM) glial response was maintained throughout the period studied. Subsets of astrocytes of different reactivities were observed, most of their elements being highly intermeshed. In 20-month-old animals, nbM lesions produced less positive, but similar, glial reactive patterns. This glial reactivity was superposed onto the glial reactivity of old age. All these results are discussed. The maintenance of reactive astrocytes many months after lesioning suggests the existence of cellular factors other than those produced by damaged nbM neurons. Taking into account the role of glial cells under pathological conditions, it is possible that these reactive astrocytes in humans could promote neurodegenerative processes, such as amyloid plaque formation and neurodegeneration (Alzheimer's disease). Along this line, nbM cholinergic involution could then originate cortical involution through induced reactive astrocytosis. PMID- 10821928 TI - Neurotoxicity after hypoxia/during ischemia due to glutamate with/without free radicals as revealed by dynamic changes in glucose metabolism. AB - Fresh rat brain slices were incubated with [18F]2-fluoro-2-deoxy-D-glucose ([18F]FDG) in oxygenated Krebs-Ringer solution at 36 degrees C, and serial two dimensional time-resolved images of [18F]FDG uptake in the slices were obtained on imaging plates. The fractional rate constant of [18F]FDG (proportional to the cerebral glucose metabolic rate) from pre-loading of ischemia (O(2) and glucose deprivation)/hypoxia (O(2) deprivation) to the reperfused/reoxygenated post loading phase was quantitatively evaluated by applying the Gjedde-Patlak graphical method to the image data. Against ischemia an N-methyl-D-aspartate antagonist and hypothermia, but not a free radical scavenger, showed a protective effect when administered during ischemia, whereas no such effect was achieved with any of the above agents when administered after reperfusion. Against hypoxia, there was no protective effect with any of the above agents when administered during hypoxia, although an effect was noted with each when administered after reoxygenation. Excitatory amino acids during ischemia loading were found to be the main factor in the neuronal damage associated with ischemia, while in hypoxia, excitatory amino acids working in tandem with free radicals immediately after reoxygenation were implicated. PMID- 10821929 TI - Late developmental expression of DARPP-32 in the chick optic tectum. AB - Immunohistochemical techniques reveal that the dopamine- and cAMP-regulated phosphoprotein DARPP-32 is detectable in neurons of the chick optic tectum starting on embryonic day 13. The expression levels then increase steadily from embryonic day 15 through the first posthatching day. After 15 days posthatching, expression of DARPP-32 reaches the adult pattern, with many labeled cells in tectal layers 11 and 12. These cells exhibit a bipolar shape, with long processes directed both to the deep and superficial layers. These results suggest that DARPP-32 is present in specific neuronal populations of the chick tectum and that this protein may not have a function in early ontogenetic processes. PMID- 10821930 TI - In vivo neurochemical effects of the acetylcholinesterase inhibitor ENA713 in rat hippocampus. AB - Oral ENA713 (0.5, 1.5 and 4.5 mg/kg), an acetylcholinesterase inhibitor (AChEI), dose-dependently enhanced extracellular acetylcholine concentrations in the hippocampus of freely moving rats. This effect was paralleled by changes in both noradrenergic and dopaminergic transmission. In particular, ENA713 significantly decreased noradrenaline concentrations, whereas it significantly increased homovanillic acid levels, without affecting dopamine concentrations. Neither serotonin nor gamma-aminobutyric acid levels were modified by ENA713. These findings extend the neurochemical profile of ENA713 and suggest that it could be useful for the treatment of Alzheimer-type dementia which is associated with multiple neurotransmitter abnormalities in the brain. PMID- 10821931 TI - Progesterone attenuates persistent inflammatory hyperalgesia in female rats: involvement of spinal NMDA receptor mechanisms. AB - The relationship between endogenous gonadal steroid levels and persistent or chronic pain is poorly understood. These studies used an inflammation model to examine the role of the gonadal steroid, progesterone, in the development of persistent pain and hyperalgesia in lactating ovary-intact and ovariectomized rats. The results indicate that constant high plasma levels of progesterone attenuate inflammatory hyperalgesia by a mechanism involving inhibition of N methyl-D-aspartate receptor activation at the spinal cord level. Since the pattern of high progesterone in lactating rats mimics the progesterone component of the luteal phase of the human menstrual cycle, these findings have significance in persistent or chronic pain conditions that are most prevalent in females. PMID- 10821932 TI - Immunohistochemical study on the distribution of neuronal voltage-gated calcium channels in the rat cerebellum. AB - Many neuronal processes are regulated by calcium influx through voltage-gated calcium channels (VGCCs), including protein phosphorylation, gene expression, neurotransmitter release, and firing patterns of action potential. In the present study, we have used anti-peptide antibodies directed against a unique sequence in rat alpha(1A), alpha(1B), alpha(1C) and alpha(1D) subunits of VGCCs to determine their cellular distribution in normal rat cerebellum. Throughout the molecular layer, immunoreactivity for alpha(1B) and alpha(1D) subunits were found in the cell bodies of basket and stellate cells as well as in the neuropil. In the Purkinje cells, only alpha(1C)-IR was observed in the dendritic branches of Purkinje cells, whereas immunoreactivity for alpha(1B) and alpha(1D) subunits were rarely found in the cell bodies of Purkinje cells. Immunoreactivity for the alpha(1A), alpha(1B,) and alpha(1D) subunits were strong in the granule cell bodies, whereas alpha(1C)-IR was not prominent in the cell bodies. In the cerebellar nuclei, a distinct band of punctate immunoreactivity for the alpha(1A), alpha(1B), alpha(1C), and alpha(1D) subunits were observed. The overall results of the above localization study showed clearly that the alpha(1A), alpha(1B,) alpha(1C) and alpha(1D) pore forming subunits of VGCCs have differential distribution in the rat cerebellum. The present studies may provide useful data for such future investigations to understand the role of calcium channels in neurological pathways. PMID- 10821933 TI - Evidence that reversed glutamate uptake contributes significantly to glutamate release following experimental injury to the rat spinal cord. AB - Released excitatory amino acids contribute significantly to secondary damage following spinal cord injury. Reversal of normal transport due to cell membrane depolarization may contribute to this release. We tested this by administering dihydrokainic acid (DHK), a non-transported glutamate uptake blocker, into the rat spinal cord by microdialysis in association with contusion spinal cord injury. Glutamate release in response to injury was reduced by 34% (P<0.05) when 3 mM DHK was administered within the microdialysis fiber, suggesting that reversed transport is an important contributor to glutamate release upon spinal cord injury. PMID- 10821934 TI - The effect of steroid sulfatase inhibition on learning and spatial memory. AB - Steroid sulfatase inhibitors can enhance the concentration of the neurosteroid DHEAS in rat brain. Previous studies have demonstrated that the steroid sulfatase inhibitor (p-O-sulfamoyl)-N-tetradecanoyl tyramine (DU-14) could reverse scopolamine induced amnesia in rats in a passive avoidance memory paradigm. The intent of this study was to determine whether chronic pretreatment with DU-14 could reverse scopolamine amnesia and/or enhance spacial memory in the place, probe and cued versions of the Morris water maze (MWM). Rats were divided into four groups and administered IP for 15 days either DU-14 (30.0 mg/Kg) or corn oil (1.0 ml/Kg) vehicle. On training days animals were administered either scopolamine (1.0 mg/Kg) or saline (1.0 ml/Kg). The groups administered DU-14 displayed a significant enhancement in learning and spacial memory in the place version of the MWM, when compared to respective vehicle-scopolamine and vehicle saline groups. In the probe version, the DU-14-saline group remained in the target quadrant of the maze significantly longer than any of the other groups indicating enhanced retention. In the cued version of the MWM, treatment with DU 14 did not significantly change escape latency suggesting that the steroid sulfatase inhibitor did not alter motivation or locomotion. These results suggest that the chronic administration of steroid sulfatase inhibitors enhance learning and spatial memory in rats. PMID- 10821935 TI - Serotonergic agents modulate antidepressant-like effect of the neurosteroid 3alpha-hydroxy-5alpha-pregnan-20-one in mice. AB - The present study demonstrated the antidepressant-like effect of neurosteroid 3alpha-hydroxy-5alpha-pregnan-20-one (3alpha, 5alpha THP) in mouse forced swim test of depression and its modulation by different serotonergic agents. Pretreatment with the selective serotonin reuptake inhibitor, fluoxetine (5 mg/kg, i.p.), the 5-HT releaser, fenfluramine (10 mg/kg, i.p.), the 5-HT(1A) receptor agonist, 8-OH-DPAT (0.1 mg/kg, s.c.), the 5-HT(1B/1C) receptor agonist, TFMPP (4 mg/kg, s.c.) and the 5-HT(2A/1C) receptor agonist, DOI (2 mg/kg, s.c.) potentiated the antidepressant-like effect of 3alpha, 5alpha THP. At these doses the serotonergic agents per se did not modify the duration of immobility. However, fluoxetine (20 mg/kg, i.p.), fenfluramine (20 mg/kg, i.p.) or imipramine (5 or 20 mg/kg, i.p.) not only reduced immobility but also enhanced the antidepressant-like effect of 3alpha, 5alpha THP. Such a potentiating effect of the 5-HT(1A) or the 5-HT(2A/1C) receptor agonist was not antagonized by the sub effective dose (0.1 mg/kg, s. c.) of their respective antagonists p-MPPI or ketanserin. Pretreatment with p-CPA (300x3 mg/kg, i.p.), a depleter of 5-HT neuronal store failed to block the influence of fluoxetine and fenfluramine on antidepressant-like effect of 3alpha, 5alpha THP. The accelerated effect of 3alpha, 5alpha THP in presence of serotonergic agents was antagonized by the GABA(A) receptor antagonist, bicuculline (1 mg/kg, i.p.) or the 3alpha hydroxysteroid oxidoreductase enzyme inhibitor, indomethacin (5 mg/kg, i.p.). These findings for the first time demonstrate that serotonergic agents potentiate the antidepressant-like action of 3alpha, 5alpha THP, by enhancing the GABAergic tone as a likely consequence of increased brain content of this neurosteroid. PMID- 10821936 TI - Increasing CNS norepinephrine levels by the precursor L-DOPS facilitate beam walking recovery after sensorimotor cortex ablation in rats(1) PMID- 10821937 TI - Modified Karhunen-Loeve expansion for evaluating skin-colour-associated melanoma risk factors. AB - An approach based on the modified Karhunen-Loeve expansion (MKLE) of constitutive and facultative skin colour data acquired by colorimeters in melanoma patients and healthy control subjects, was used to identify two colour features defining skin-colour-associated risk of melanoma. None of four common statistical classifiers trained on colour features were sufficiently accurate for allowing skin colour alone to be used for classification purposes, though a Bayesian quadratic classifier matched the transformed data well. This study supersedes the indeterminate character of most common clinical criteria based on qualitative factors and, irrespective of the results of classification, provides objective skin colour information for the prevention of melanoma. PMID- 10821938 TI - Simulation of cardiac excitation patterns in a three-dimensional anatomical heart atlas. AB - Computerized anatomical atlas systems enable interactive investigation of digital body models. Here we present a three-dimensional atlas of the human heart, based on image data provided in the Visible Human Project. This heart atlas consists of multiple kinds of cardiac tissues and offers unlimited possibilities for its visual exploration. A temporal dimension is added to the underlying heart model by simulation of cardiac excitation spreading. For this purpose a second generation cellular automata algorithm is adapted to the excitation kinetics of cardiac tissue. The presented system is shown as a successful method for the visualization-based investigation of cardiac excitation. PMID- 10821939 TI - Numerical approach to plug-flow activated sludge reactor kinetics. AB - In this study, general relationships applicable to a wide variety of microbial mediated treatment processes are developed using a number of continuous-flow stirred tank reactors-in-series. Simultaneous equations resulting from mass balance on substrate and biomass are solved numerically taking the longitudinal biomass gradient into account. A relationship between substrate and biomass concentrations is developed as a function of hydraulic residence time. Monod kinetics and mathematical models of reactors-in-series are used to represent the actual conditions resulting from varying degrees of axial dispersion and wastewater quality. Dimensionless quantities are used to reduce the number of parameters to be taken into account. Computer techniques are applied to express the results generally. By means of the numerical approach, the variation of the ratio between the hydraulic residence time of a given reactor and that of an equivalent plug-flow tank with the same inlet and outlet conditions is investigated. Results are expressed in the form of graphs to characterize the plug-flow activated sludge systems and to provide a basis of design. Experimental data reported in the literature are also evaluated to demonstrate the cases where the existing classical solution to the problem differs from the actual results obtained from the computer program and the design graphs given. PMID- 10821940 TI - A java-based application for differential diagnosis of hematopoietic neoplasms using immunophenotyping by flow cytometry. AB - We describe the implementation of a Java-based application for differential diagnosis of hematopoietic neoplasms using immunophenotyping by flow cytometry. The current version of this Java applet includes the knowledge-base for 33 hematopoietic neoplasms and 43 diagnostic immunophenotyping markers. Java, a new object-oriented computing language, helps facilitate development of this applet, a platform-independent module that can be implemented on the World Wide Web. As the Web rapidly becomes more accessible to users around the world, Web-based software may eventually form the core of decision-support systems in clinical settings. Java-based applications, such as the one described in this paper, are expected to contribute significantly in this area. PMID- 10821941 TI - A new method to determine a fractal dimension of non-stationary biological time serial data. AB - We devised a new analysis using quartile deviation of integrated and subtracted fluctuation, termed QIS-A, to determine a fractal dimension of non-stationary fluctuation. In the algorithm, computations of the quartile deviation, Q(n), of all integrated and subtracted fluctuations are repeated over all scales (n). The fractal scaling exponent is determined as a slope of the line relating log Q(n) to log n. Comparison of the QIS-A and a spectral analysis using 20 computer simulated fractional Brownian motions demonstrates robustness of the QIS-A to non stationary fluctuations. PMID- 10821942 TI - The use of Tween 20 in immunoblotting assays for the detection of autoantibodies in connective tissue diseases. AB - Autoantibodies directed against intracellular antigens can be detected by immunoblotting (IB). Due to its high sensitivity this technique has many advantages, but it can give misleading results when the specific bands are weak or blurred against the background staining. To decrease background staining, non ionic detergents (Tween 20, Triton X-100, Nonidet P-40) are generally used as blocking agents. Moreover, these agents appear to have a renaturating action towards proteins and antigens. Tween 20 has a more pronounced renaturating effect on proteins than other detergents and thereby improves antigen-antibody binding. To evaluate the effect of Tween 20 on specific autoantibody detection by IB, we tested the sera of 162 patients with connective tissue diseases (CTDs) by adding this detergent at certain steps of the IB assay. We found that the use of Tween 20 in the IB procedure significantly improved the binding of autoantibodies to Jo 1, Scl70, (U1)RNP 68 kDa and C, Sm B/B' and D. Moreover, it increased the sensitivity for the detection of anti-Sm D peptide in systemic lupus erythematosus (SLE) sera with no decrease in specificity. In contrast, the addition of Tween 20 significantly decreased the binding of autoantibodies specific for ribosomal P proteins, La/SSB, Ro/SSA, but not the overall sensitivity and specificity of the method. We conclude that the addition of Tween 20 to standard IB is advantageous for anti-nuclear antigen antibody detection and improves the sensitivity of the method in revealing anti-Sm-positive sera in SLE. However, Tween 20 is not recommended for the detection of anti-cytoplasmic antibodies. PMID- 10821943 TI - Loss of CD34(+) hematopoietic progenitor cells due to washing can be reduced by the use of fixative-free erythrocyte lysing reagents. AB - Current protocols for sample preparation before flow cytometric enumeration of CD34(+) hematopoietic progenitor cells (HPC) include both lyse-non-wash and lyse and wash methods. Erythrocyte lysis without washing is the method of choice when absolute cell counts are to be assessed, whilst a washing step is recommended for immunological subtyping of CD34(+) cells in order to reduce background fluorescence. Here, we analyzed the effect of the interaction between type of erythrocyte lysis reagent and washing on the outcomes of (i) CD34(+) cell enumeration and (ii) expression of CD38 by CD34(+) cells in a single-platform, whole-blood staining assay [Gratama, J.W., Keeney, M., Sutherland, D.R., 1999. Enumeration of CD34(+) hematopoietic stem cell and progenitor cells. Curr. Protocols Cytometry 6(4), 1-22.]. We studied seven commercially available lysing reagents (five containing fixative and two fixative-free) using 12 samples from cord blood (n=4), mobilized peripheral blood (n=4) and apheresis products (n=4). Using the lyse and wash technique, significant reductions of absolute and relative numbers of CD34(+) cells, as well as in the numbers of lymphocytes and leukocytes, were observed on samples that had been lysed using fixative containing buffers as compared to the lyse-no-wash technique. Cell losses due to washing could be significantly reduced when samples were lysed using fixative free buffers. 'Postfixation' using PBS+1% paraformaldehyde of samples that had been lysed using fixative-free buffers and then washed did not result in additional loss of CD34(+) cells or other cell types. Finally, washing unfixed samples led to a slight decrease of CD38 monoclonal antibody bound to CD34(+) cells as compared to samples that had been fixed during erythrocyte lysis. These results indicate that fixation renders (CD34(+)) cells sticky and leads to their loss from the cell suspension upon centrifugation and resuspension. We conclude that all seven lysing reagents can be used with confidence in a lyse-no-wash technique, but that only fixative-free lysing reagents should be used when a washing step is considered necessary. PMID- 10821944 TI - Stereological quantification of carboxyfluorescein-labeled rat lung metastasis: a new method for the assessment of natural killer cell activity and tumor adhesion in vivo and in situ. AB - The function of natural killer (NK) cells is often studied by assessing in vitro levels of NK cell mediated lysis of target cells, or by assessing in vivo levels of lung tumor cell retention or metastatic colonization of intravenously injected tumor cells. However, these methods do not permit direct quantification and visualization of NK cells and their targets in vivo and in situ. Here, a new approach is described to visualize effector-to-target interactions as well as to estimate total numbers of targets in the lung, in vivo and in situ. MADB106 tumor cells were vitally labeled using carboxyfluorescein (CFSE) and intravenously (i.v.) injected into Fischer 344 rats (10(6) cells/rat). This mammary adenocarcinoma derived cell line is syngeneic to the inbred Fischer 344 rat and highly sensitive to NK cell activity in vivo. Effector-to-target interactions were visualized by immunostaining. Using the optical fractionator method, total numbers of CFSE-labeled MADB106 tumor cells were estimated in the left lung of the animals 5 min after tumor inoculation. To further demonstrate the usefulness of this approach in reflecting in vivo processes, rats were inoculated with MADB106 cells and simultaneously with a single i.v. bolus of either 1 microg/kg adrenaline or saline. Both lungs were removed 5 min later. Adrenaline caused a significant 80% reduction in the total number of lung CFSE-labeled MADB106 tumor cells, suggesting a rapid modulation of metastasis by stress hormones. This new approach facilitates the monitoring of effector-to-target interactions and the quantification of immune cell function or tumor adhesion in vivo and in situ. PMID- 10821945 TI - A novel method for the simultaneous assessment of natural killer cell conjugate formation and cytotoxicity at the single-cell level by multi-parameter flow cytometry. AB - A flow cytometric assay for the combined measurement of cell-mediated cytotoxicity and conjugate formation has been developed. Cytolysis is detected by propidium iodide uptake. Target cells, effector cells and conjugates between targets and effectors are separated by post-culture immunophenotyping and their scatter profiles. Pre-assay staining of cells is thus not required. Each cluster of cells can be further examined at the single-cell level by simultaneously performed additional immunophenotyping. Two applications were established: the assessment of NK cell activity against K562 cells and the evaluation of LAK cell cytotoxicity against both K562 and Daudi cells. A comparison with the standard 51Cr release assay for the detection of NK cytotoxicity showed that the two assays were strongly correlated, but the sensitivity of the flow cytometric assay was significantly higher. PMID- 10821946 TI - Neutrophil products and alterations in epithelial junctional proteins: prevention of artifactual degradation. AB - Recent reports of disruption of endothelial cell adherens junction proteins during neutrophil adhesion and transmigration have been challenged as being partly due to post-fixation artifactual release of neutrophil-derived proteases. In this study we examined alterations in the epithelial junctional complex during neutrophil adhesion. Using standard fixation protocols, neutrophil addition to epithelial monolayers resulted in gross disruption of apical junction protein immunofluorescence. However, the inclusion of a post fixation incubation step with formic acid resulted in epitope preservation. These observations indicate that neutrophil derived products, likely proteases, remain active despite prolonged exposure to conventional fixatives. This may result in diffuse and artifactual loss of epithelial junctional protein immunofluorescence. Formic acid prevents this loss of epitope staining and may be considered as an agent to preserve protease-sensitive endothelial or epithelial immunoreactivity. PMID- 10821947 TI - Measurement of specific IgA in faecal extracts and intestinal lavage fluid for monitoring of mucosal immune responses. AB - Currently available methods for the evaluation of antigen-specific immune responses in the intestine, i.e. measurement of IgA in intestinal lavage and antibody secreting cells (ASC) in peripheral blood, are not applicable to large scale immunogenicity studies or to kinetic studies where repeated sampling is required. Simple and reliable methods need to be developed. Intestinal lavage and faecal samples were collected from 12 mice on days 0, 14, 21, 28 and 35 following initial immunization with four doses of cholera toxin (CT) by the gastric or rectal routes. The concentrations of anti-CT IgA in the faecal extracts showed a high level of correlation with those in the lavage samples (Spearman's correlation coefficient=0.85, P<0. 0001) regardless of the route of CT administration. Moreover, the kinetics of the immune response as reflected in the faecal extracts mirrored those in the lavage samples regardless of immunization route. As compared to gastric immunization, rectal administration of CT yielded higher levels of anti-CT IgA in both intestinal lavage fluids and in faecal extracts. The use of rectal immunization and the measurement of IgA in faecal extracts for monitoring mucosal immune responses may be relevant for the development of effective enteric vaccines. PMID- 10821948 TI - Dissection of an antibody paratope into peptides discloses the idiotope recognized by the cognate anti-idiotypic antibody. AB - Using methods of parallel synthesis, the complete amino acid sequence of an Ab 1 antibody (Tg 10, an anti-human thyroglobulin monoclonal antibody) was made in the form of a set of 100 synthetic overlapping peptides. This set of immobilized peptides was allowed to react with the cognate Ab2 (AI 10, a highly purified rabbit anti-idiotypic polyclonal antibody to Tg 10). A dominant peptide idiotope, INTFSGVPTYA, was thus mapped, which corresponds mainly to the CDR2 region from the V(H) domain of the Tg 10 mAb. A synthetic peptide replica of this idiotope was found to bind to AI 10 with an affinity (K(D) in the 10(-8) M range, as measured using BIACORE technology) which represents a significant part of the affinity of the complete Tg 10 antibody (K(D) in the 10(-9) M range). The synthetic peptide also elicited anti-idiotypic antibodies in rabbits that recognized specifically the Ab1 antibody in an Ab1- and antigen-inhibitable manner. The peptide idiotope was further characterized chemically by the identification of residues important for binding to the Ab2 and by modelization of its structure. Our approach makes it readily possible to map and characterize functional, continuous-type idiotopes that could be further used to manipulate the immune response by peptide technologies. PMID- 10821949 TI - ELISA to evaluate plasma anti-asparaginase IgG concentrations in patients with acute lymphoblastic leukemia. AB - The development of antibodies to asparaginase may attenuate the pharmacologic effect of asparaginase treatment, may be associated with hypersensitivity reactions, and may necessitate switching to a different commercial asparaginase preparation for current or future therapy. Thus, development of an ELISA for measurement of anti-asparaginase antibody levels is important in the clinical setting. An anti-asparaginase antibody reference was established by screening 65 plasma samples from six patients with acute lymphoblastic leukemia (ALL) who had recently developed a hypersensitivity reaction to Escherichia coli or Erwinia chrysanthemi asparaginase therapy. Twenty-one plasma samples were selected for the anti-asparaginase antibody reference pool. Five micrograms per milliliter of commercial E. coli and Erwinia asparaginase and 10 microg/ml of E. coli asparaginase conjugated with polyethylene glycol (PEG asparaginase) were found to be optimal as coating antigen concentrations. Anti-asparaginase antibody concentrations were determined using a commercial polyclonal goat anti-human IgG horseradish peroxidase conjugate. The antibody reference curves were linear in a range of absorbance from 0.1 to 1. 5 O.D. units for dilutions from 1:1600 to 1:51,200. Inter-assay coefficients of variation were 9.04, 14.7 and 13.0%, and intra-assay coefficients of variation were 1.44, 4.43 and 3.28% for antibodies against E. coli, Erwinia, and PEG L-asparaginase, respectively. The cut-off for positivity in plasma was determined as mean+2 S.D. of the optical density values for plasma from untreated healthy volunteers. Measurement of specific IgG by this ELISA allows for the evaluation of plasma anti-asparaginase antibody concentrations in patients receiving one or more of the multiple commercial L asparaginase preparations. PMID- 10821950 TI - A highly sensitive, non-radioactive assay for T cell activation in cattle: applications in screening for antigens recognised by CD4(+) and CD8(+) T cells. AB - We describe a highly sensitive, non-radioactive assay for T cell activation, based on the rapid induction of class II MHC expression by constitutively negative bovine endothelial cells, when cultured in the presence of supernatants derived from activated bovine T cells. We demonstrate the effectiveness of this assay in detecting rBoIFNgamma and activation of immune CD4(+) and CD8(+) T cell lines and clones in response to specific antigen and transfected COS-7 cells, respectively. We also demonstrate its utility in identifying purified pathogen fractions that activate immune CD4(+) T cell clones. PMID- 10821951 TI - A two-step culture method starting with early growth factors permits enhanced production of functional dendritic cells from murine splenocytes. AB - Dendritic cells (DC) are professional antigen presenting cells (APC) able to activate naive T cells and initiate the immune response. They are present in most tissues at very low concentrations and are difficult to isolate. DC can be obtained in larger numbers by their propagation from progenitors present in blood, bone marrow and spleen. However, biochemical studies and biological analysis of DC functions require very large numbers of these cells. In this paper, we described a two-step culture system using unfractionated splenocytes from BALB/c mice as a source of DC progenitors. The proliferative capacity of the progenitors is amplified in the first step of the culture (day 0-6) using different combinations of early acting cytokines combined or not with granulocyte macrophage CSF (GM-CSF). The second step of the culture starts at day 6 with the removal of early growth factors in order to allow the differentiation and final maturation of DC during 2-3 weeks of culture with flt-3 ligand (flt-3L) and GM CSF. The addition of Stem Cell Factor (SCF) or IL-6 to the standard combination of flt-3L+/-GM-CSF produces a large increase in the proliferation of GM and DC progenitors (28 times and 11 times respectively) in the first step of the culture. This proliferative wave of DC progenitors is followed by the production of a high percentage of immature and mature DC in flt-3L+GM-CSF stimulated cultures. The best combination of early cytokines in terms of proliferative activity and subsequent level of DC production was flt-3L+IL-6+GM-CSF, which permitted the generation of 1 to 2x10(9) DC from one single spleen. Using this growth factor cocktail, a mixture of immature (2/3) and mature (1/3) DC was produced until day 14 of culture, and levels of MHC class II and costimulatory molecules (CD40, B7.2) increased between 2 and 4 weeks of incubation, or within 2 days when stimulated by IL-4 or LPS. The splenic DC produced after 2 weeks of culture are fully functional, exhibiting a high capacity of endocytosis when immature, a strong stimulatory reactivity in mixed leukocyte reaction and consistently producing high levels of bioactive IL-12 p70 after CD 40 ligation in the presence of LPS between 13 and 43 days of culture. PMID- 10821953 TI - An on-line quantitative immunoassay system based on a quartz crystal microbalance. AB - We have established a QCM immunoassay system which allows on-line and quantitative monitoring throughout the entire detection procedure and provides information on the surface coverage and the binding ratio of antibody to antigen. Compared to conventional immunoassay systems the QCM system offers advantages of short response times, obviates the need for additive labeling reagents, and permits direct conversion of a frequency signal into mass accumulation. PMID- 10821952 TI - Effects of fluorescent dyes on selectin and integrin-mediated stages of adhesion and migration of flowing leukocytes. AB - Fluorescent dyes assist visualisation of leukocytes for intravital studies of adhesion or for in vitro studies utilising whole blood. We have used in vitro flow-based assays to investigate the effects of three fluorescent dyes (acridine orange, AO, 5-100 microg/ml; calcein-AM, C-AM, 5-20 microg/ml; rhodamine 6G, R6G, 10-100 microg/ml) on adhesion and migration of isolated neutrophils and mononuclear cells. AO had little effect on the number or velocity of neutrophils rolling on P-selectin presented by a surface coated with platelets. However, AO did cause a dose- and time-dependent conversion of rolling to immobilisation. Pretreatment of neutrophils with an antibody against CD18 prevented this conversion to stationary adhesion, indicating that beta(2) integrins were activated by AO. C-AM had little effect on neutrophil behaviour, but tended to cause some immobilisation at the highest concentration. R6G did not affect the number of neutrophils that bound to the platelet monolayer or the percentage rolling, but the rolling velocity of the neutrophils was increased in a dose dependent manner. None of the dyes impaired the ability of neutrophils to respond to formyl peptide by converting from rolling to stationary adhesion. Neither C-AM nor R-6G reduced the number of flowing neutrophils or mononuclear cells binding to endothelial cells stimulated with tumour necrosis factor. Interestingly, R-6G inhibited transendothelial migration of mononuclear cells but not neutrophils, while C-AM did not affect transmigration of either cell type. The dose-dependent effects of dyes should be taken into consideration when designing any experimental protocol. AO does not appear to be a suitable dye for adhesion studies. R6G and C-AM can be used at approximately 10 microg/ml (a concentration at which cells can be clearly visualised) although R-6G specifically inhibits the migratory response of mononuclear cells. PMID- 10821954 TI - A novel mammalian expression screen exploiting green fluorescent protein-based transcription detection in single cells. AB - The accumulation of DNA sequence information from large-scale genomic and random library sequencing projects is leading to the rapid identification of many putative genes, virtual transcripts and ESTs of unknown function. There is therefore an increasing need for high throughput, sensitive and robust methods for identification and characterisation of genes, and/or their products, based on function. We describe a high throughput functional expression screen based on semi-quantitative analysis of enhanced green fluorescent protein expression in single cells by confocal microscopy. The assay was implemented in a micro-scale format, requiring around 10(4) cells/test. The system was validated by co transfection of a series of cDNAs encoding pro-inflammatory cytokine intracellular signal mediators with a d2EGFP reporter containing a cytokine responsive promoter. The majority of the test plasmids gave a detectable signal above background at a pool size of 250-500. Replicate tests indicate that the assay is reproducible at this pool size. At this level we demonstrate that large (>10(6) transformants) libraries can be feasibly screened. PMID- 10821955 TI - In vivo selection of sFv from phage display libraries. AB - The development of phage display technology has facilitated the development of many new and sometimes novel antibody based reagents for scientific research. However, present methods for selection from phage-sFv display libraries are limited to selection against purified antigens or ex vivo cells of known origin and phenotype. Existing methods therefore preclude the isolation of sFv against unknown molecules in their natural environment, where expression is complex and subject to diverse control mechanisms. Since such a complex environment is difficult to mimic in vitro, the development of an in vivo selection procedure would greatly enhance the selection from phage display antibody libraries and lead to the development of reagents against cell surface molecules in their natural environment. This would be particularly advantageous for isolation of sFv against vascular endothelium which can readily change phenotype when cultured and is believed to express molecules in a tissue specific manner and in response to different stimuli. We describe here the development of an in vivo selection procedure in the mouse and demonstrate its potential for the selection of sFv from a phage-sFv library. The target antigen for one sFv is expressed solely on the thymic endothelium, while the second, a 165-170 kDa molecule in present on both thymic endothelium and the perivascular epithelium. PMID- 10821956 TI - Functional human monoclonal antibodies of all isotypes constructed from phage display library-derived single-chain Fv antibody fragments. AB - We have constructed a series of eukaryotic expression vectors that permit the rapid conversion of single chain (sc) Fv antibody fragments, derived from semi synthetic phage display libraries, into intact fully human monoclonal antibodies (mAb) of each isotype. As a model, a scFv fragment specific for sheep red blood cells (SRBC) was isolated from a semi-synthetic phage antibody (Ab) display library, and used to produce human mAbs of IgM, IgG1-IgG4, IgA1, IgA2m(1) and IgE isotype in vitro in stably transfected cells. N-terminal protein sequence analysis of purified immunoglobulin heavy (H) and light (L) chains revealed precise proteolytic removal of the leader peptide. Biochemical analysis of purified recombinant human mAbs demonstrated that properly glycosylated molecules of the correct molecular size were produced. The IgG and IgA mAbs retained SRBC binding activity, interacted with different Fc receptor-transfectants, and induced complement-mediated hemolysis and Ab-dependent phagocytosis of SRBC by neutrophils in a pattern consistent with the immunoglobulin (Ig) H chain isotype. We conclude that in vitro produced recombinant human mAbs constructed from phage display library-derived scFv fragments mirror their natural counterparts and may represent a source of mAbs for use in human therapy. PMID- 10821957 TI - Gene expression profiling in human peripheral blood mononuclear cells using high density filter-based cDNA microarrays. AB - Microarray technology has provided the ability to analyse the expression profiles for thousands of genes in parallel. The need for highly specialised equipment to use certain types of microarrays has restricted the application of this technology to a small number of dedicated laboratories. High-density filter-based cDNA microarrays provide a low-cost option for performing high-throughput gene expression analysis. We have used a model system in which filter-based cDNA microarrays representing over 4000 known human genes were used to monitor the kinetics of gene expression in human peripheral blood mononuclear cells (PBMCs) stimulated with phytohaemagluttinin (PHA). Using software-based cluster analysis, we identified 104 genes that altered in expression levels in response to PHA stimulation of PBMCs and showed that there was a considerable overlap between genes with similar temporal expression profiles and similar functional roles. Comparison of microarray quantitation with quantitative PCR showed almost identical expression profiles for a number of genes. Coupled with the fact that our findings are in agreement with a large number of independent observations, we conclude that the use of filter-based cDNA microarrays is a valid and accurate method for high-throughput gene expression profiling. PMID- 10821958 TI - A discrete-event computer model of feline herpes virus within cat populations. AB - The purpose of this work was to study the epidemiology of feline herpes virus (FHV), which causes a respiratory disease within natural populations of domestic cats. A stochastic model was constructed using discrete-events simulation. Two habitats (rural vs. urban) were simulated, featuring different demographic, spatial and social patterns. The evolution of immunity in individuals was reproduced, allowing for the random recrudescence of latent infections (influenced by environment and reproduction). Hypotheses concerning the circulation of FHV were examined regarding the role of host density and the possibility of reinfection of host. Uncertainty analyses were performed on the basis of replicated Monte Carlo sampling. The results were in good agreement with serologic data from a long-term study conducted on five populations in France. The model satisfactorily reproduced the variability of natural immunity, and the epidemic features observed. The simulations have shown that FHV can persist in small populations (because of its capacity of reactivation leading to epidemics). However, the impact on demography was not dramatic. The most important parameters in determining change in epidemiology of FHV were: transmission rate corresponding to 'friendly' contacts, and the recrudescence rate of FHV. However, an interaction between these two parameters did not allow estimation of their values. PMID- 10821959 TI - Suboptimal reproductive performance of dairy cattle kept in smallholder herds in a rural highland area of northern Tanzania. AB - The objectives of the present study were to assess the reproductive performance and cumulative incidence risk of reproductive disorders and to compare the success of artificial insemination (AI) to natural service (by handmating; NS) in dairy cattle kept in smallholder herds under a zero-grazing system in a rural highland area of Tanzania. Data on occurrence of all normal and abnormal reproductive events were collected for 215 adult animals belonging to 74 households. The median and range of the intervals: intercalving, calving to first service, and calving to pregnancy were 477 (335-860), 154 (38-486) and 206 (61 567) days, respectively. Breed and parity did not affect the reproductive parameters (P>0.05). However, cows in the milked group had a shorter median calving interval than those in the suckled group (P<0.001). The overall percentage pregnant and the percentage pregnant to first service were higher in the NS than in the AI group (49 vs. 32%; P=0.007) and (67 vs. 25%; P<0.001), respectively. The median numbers of services per pregnancy were not different between the AI (3) and NS (2) groups (P=0.17). The cumulative incidence risk of abortion, dystocia, prolapse, retained fetal membranes, mastitis, milk fever and cyclic non-breeders were 16.0, 1.7, 2.5, 4.2, 5.0, 1.7, and 6.1%, respectively. Hoof overgrowth (4.6%) and hoof deviation (4.6%) were the most-frequent digital problems. We concluded that reproductive parameters and cumulative incidence risk of abortion show suboptimal reproductive performance in rural-based, zero-grazed smallholder dairy herds in Tanzania particularly those using AI. PMID- 10821960 TI - Use of Brucella abortus vaccine strain RB51 in pregnant cows after calfhood vaccination with strain 19 in Argentina. AB - One hundred and seven pregnant cows, which had been calfhood vaccinated with Brucella abortus strain 19 (S-19) were revaccinated with either S-19 or strain RB51 (S-RB51). All S-19-revaccinated animals seroconverted, while none of the RB51-revaccinated animals seroconverted. Two out of 25 (8%) S-19-revaccinated animals aborted, while none of the 57 RB51-revaccinated group aborted. Four of the S-19-revaccinated animals shed S-19 in the milk for at least 7 days, while only 1 cow shed S-RB51 for at least 3 days (but <7 days) post-parturition. Revaccination of strain 19 calfhood-vaccinated, pregnant cattle with S-RB51 appears to be a safe procedure with no diagnostically negative consequences. PMID- 10821961 TI - Medicinal plants used for dogs in Trinidad and Tobago. AB - This paper documents ethnoveterinary medicines used to treat dogs in Trinidad and Tobago. In 1995, a 4-stage process was used to conduct the research and document the ethnoveterinary practices. Twenty-eight ethnoveterinary respondents were identified using the school-essay method, which is a modified rapid rural appraisal (RRA) technique. Semi-structured interviews were held with these respondents as well as with 30 veterinarians, 27 extension officers and 19 animal health assistants and/or agricultural officers, and the seven key respondents that they identified. The final step involved hosting four participatory workshops with 55 of the respondents interviewed to discuss the ethnoveterinary data generated from the interviews and to determine dosages for some of the plants mentioned. Supplementary interviews were conducted in 1997 and 1998. Seeds of Carica papaya, and leaves of Cassia alata, Azadirachta indica, Gossypium spp., Cajanus cajan and Chenopodium ambrosiodes are used as anthelmintics. The anthelmintics Gossypium spp. and Chenopodium ambrosiodes are the most frequently used species. Crescentia cujete pulp, Musa spp. stem exudate, the inside of the pods of Bixa orellana, leaves of Cordia curassavica and Eclipta alba plant tops are used for skin diseases. Musa spp. stem exudate, seeds of Manilkara zapota, Pouteria sapota and Mammea americana and leaves of Cordia curassavica, Scoparia dulcis and Nicotiana tabacum are used to control ectoparasites. Dogs are groomed with the leaves of Cordia curassavica, Bambusa vulgaris and Scoparia dulcis. Psidium guajava buds and leaves and the bark of Anacardium occidentale are used for diarrhoea. Owners attempt to achieve milk let-down with a decoction of the leaves of Stachytarpheta jamaicensis. The plant uses parallel those practised in human folk medicine in other Caribbean countries and in other tropical countries. PMID- 10821963 TI - Prevalence and distribution of gastro-intestinal helminths and haemoparasites in young scavenging chickens in upper eastern region of Ghana, West Africa. AB - We conducted a cross-sectional study to determine the prevalence and species of gastro-intestinal helminths and haemoparasites in 100 chickens kept under extensive management systems in Ghana, West Africa. All the examined chickens (100%) were infected with gastro-intestinal helminths; a total of 18 species were detected. The species and their prevalences were: Acuaria hamulosa (25%), Allodapa suctoria (20%), Ascaridia galli (24%), Capillaria spp. (60%), Choanotaenia infundibulum (13%), Gongylonema ingluvicola (62%), Heterakis gallinarum (31%), H. isolonche (16%), Hymenolepis spp. (66%), Raillietina cesticillus (12%), R. echinobothrida (81%), R. tetragona (59%), Strongyloides avium (2%), Subulura strongylina (10%), Tetrameres fissispina (58%), Trichostronygylus tenuis (2%), and finally one unidentified acanthocephalan (1%) and one unidentified trematode (1%). Thirty-five per cent of the chickens were infected with the haemoparasites Aegyptinella pullorum and Plasmodium juxtanucleare (prevalences 9% and 27%, respectively). Association between chicken sex and prevalences was not significant. An over-dispersed distribution was seen for most of the helminth species. PMID- 10821962 TI - A longitudinal study of serological patterns of respiratory infections in nine infected Danish swine herds. AB - Sixteen litters of seven pigs from each of nine Danish farrow-to-finish herds were followed to investigate the serological patterns caused by natural infection with Mycoplasma hyopneumoniae, Pasteurella multocida toxin and Actinobacillus pleuropneumoniae serotypes 2, 5-7, 12. In seven of the herds, pigs were followed as two separate cohorts started 4 weeks apart, and in two herds only one cohort was followed.A total of 999 pigs were included in the study. The pigs were blood sampled at weaning and subsequently every fourth week until slaughter. All pigs were examined for antibodies against M. hyopneumoniae (enzyme-linked immunosorbent assay), P. multocida toxin (enzyme-linked immunosorbent assay) and A. pleuropneumoniae serotypes 2, 5-7, 12 (complement-fixation tests). The most common pattern (28%) of seroconversion was that of pigs first seroconverting to A. pleuropneumoniae serotype 2, followed by seroconversion to M. hyopneumoniae. Each herd had a dominant serotype of A. pleuropneumoniae to which most pigs seroconverted. Seroconversion to the respiratory pathogens occurred mainly in the growing-to-finishing units (8-24 weeks). The risk of seroconversion to the P. multocida toxin was very low (<20%) and occurred late.None, four and seven herds tested seropositive to PRRS and to swine influenza virus subtypes H3N2 and H1N1, respectively, when testing 10 pigs per herd (selected randomly among the study pigs) at the age of 20 weeks. PMID- 10821964 TI - Estimates of within-herd incidence rates of Mycobacterium bovis in Canadian cattle and cervids between 1985 and 1994. AB - We analysed the individual-animal data from six of the nine outbreaks of tuberculosis in Canadian cattle and cervids from 1985 to 1994. A "positive/reactor" animal was one which had either a positive culture or a positive or suspicious reaction on a mid-cervical, comparative cervical, or gross or histopathological test for tuberculosis. Individual-animal data were collected only for herds which had one or more positive/reactor animals. Data were collected from the outbreak records in the Regional or District offices of Agriculture and Agri-food Canada's Animal and Plant Health Directorate. The within-herd spread of Mycobacterium bovis was studied by determining the most likely date at which the herd was first exposed to M. bovis and the number of reactions which had developed by the time the herd was investigated. The animal time units at risk in the herd were probably overestimated, resulting in conservative estimates of the within-herd incidence rates. Negative-binomial regression was used to investigate factors which might have influenced the within herd spread of tuberculosis. Increasing age appeared to be a risk factor for being a positive/reactor animal. When compared to animals 0-12 months old, animals 13-24 months old had an incidence rate ratio (IRR) of 7.6, while animals >24 months old had an IRR of 10.4 (p=0.009). Actual and predicted incidence rates for tuberculosis in mature (>24 months old) animals were calculated. Actual and predicted incidence rates were similar for cervids, within an outbreak. There was more variability between actual and predicted rates in the dairy and beef animals. In the one outbreak (Ontario) where there were positive/reactor cervid, dairy and beef herds, the actual incidence rate for cervids (IR=9.3 cases per 100 animal-years) was almost twice that of dairy cattle (IR=5.0) and three times that of beef cattle (IR=3.1). PMID- 10821965 TI - Techniques for analysis of disease clustering in space and in time in veterinary epidemiology. AB - Techniques to describe and investigate clustering of disease in space - the nearest-neighbour test, autocorrelation, Cuzick-and-Edwards' test and the spatial scan statistic - and in time - the Ederer-Myers-Mantel test and the temporal scan statistic - are reviewed. The application of these techniques in veterinary epidemiology is demonstrated by the analysis of a data set describing the occurrence of blowfly strike - both body strike and breech strike - between August 1998 and May 1999 in 33 commercial sheep flocks located within two local government areas of southeastern Queensland, Australia. By applying a combination of these methods, the occurrence of blowfly strike in the study area is well characterised in both space and time. Guidelines for investigating disease clusters in veterinary epidemiology are discussed. PMID- 10821966 TI - Prevalence of bovine herpesvirus-1 in the Belgian cattle population. AB - The national bovine herpesvirus 1 (BHV-1) seroprevalence (apparent prevalence) in the Belgian cattle population was determined by a serological survey that was conducted from December 1997 to March 1998. In a random sample of herds (N=556), all cattle (N=28478) were tested for the presence of antibodies to glycoprotein B of BHV-1. No differentiation could be made between vaccinated and infected animals, because the exclusive use of marker vaccines was imposed by law only in 1997 by the Belgian Veterinary Authorities. Twenty-one percent of the farmers vaccinated continuously against BHV-1. In the unvaccinated group, the overall herd, individual-animal and median within-herd seroprevalences were estimated to be 67% (95% confidence interval (CI)=62-72), 35.9% (95% CI=35.0-36.8) and 33% (quartiles=14-62), respectively. Assuming a test sensitivity and specificity of 99 and 99.7%, respectively, the true herd, individual-animal and median within herd prevalence for the unvaccinated group of herds were estimated to be 65, 36 and 34%, respectively. The true herd prevalence for dairy, mixed and beef herds were respectively, 84, 89 and 53%; the true individual-animal prevalence for those types of herds were, respectively, 35, 43 and 31%; whereas, the true median within-herd prevalences were 36, 29 and 38%. PMID- 10821967 TI - Preliminary findings of Salmonella spp. in captive green iguanas (Iguana iguana) and their environment. AB - Captive reptiles are routinely identified as reservoirs of Salmonella spp. and reports of reptile-associated salmonellosis are increasing. Unfortunately, little is known about the epidemiology of Salmonella spp. and green iguanas. We did a limited survey of a green-iguana farm in El Salvador to identify sources of Salmonella spp. in green iguanas and their environment. A limited number of samples for microbiological culture were collected from iguanas (adult, hatchling, and embryos) and their environment (food, water, soil, shelter, insects, and wild-caught lizards). Salmonella spp. was isolated from the intestine of both adult (3/20) and hatchling iguanas (8/20). There was no evidence of Salmonella spp. in the reproductive tracts of female iguanas (0/10). Salmonella spp. was isolated from the surface of 40% (7/16) of the egg surfaces tested. Salmonella spp. was not identified from the externalized yolk-sac of the iguana embryos tested. Soil samples from a breeding pen and a nest were both positive for Salmonella spp. Eight different Salmonella spp. serotypes were identified in this survey. These results suggest that horizontal transmission of Salmonella spp. is a potential source of exposure to hatchling iguanas at this facility. PMID- 10821968 TI - Factors associated with intersucking in Swiss dairy heifers. AB - Intersucking is a problem that may lead to udder damage, mastitis, milk loss, and culling of breeding animals. To analyse the risk factors for intersucking in dairy heifers in Switzerland, we asked 130 randomly selected dairy farmers about a broad spectrum of environmental factors that might be associated with intersucking (such as housing conditions, management, and the feeding of calves and heifers). In total, 2768 heifers (Swiss Brown Cattle, Simmental and Holstein Friesian) were included of which 303 had ever performed intersucking. Data at the farm level were analysed using path analysis (linear and logistic regression). Two outcome variables were considered: the occurrence of intersucking on the farm and the proportion of intersucking heifers above the cut-off of 7.2%. Farms where calves had no access to barnyard or pasture, where calves were reared in pens in enclosed buildings, where heifers after weaning were not restrained while feeding, where heifers after weaning received <0.5kg concentrate per day, and where heifers after weaning were fed > or =40% maize silage (dry-matter ratio) were most likely to have intersucking heifers. PMID- 10821969 TI - New initiatives for the control of Japanese encephalitis by vaccination: minutes of a WHO/CVI meeting, Bangkok, Thailand, 13-15 October 1998. AB - Japanese encephalitis (JE) is a leading cause of viral encephalitis in Asia that, in several countries, has been controlled effectively through national vaccine programs. However, in recent years, transmission has been recognized or has intensified in new locations where the available vaccines are either unaffordable or unlicensed. In addition, the near-eradication of poliomyelitis from Asia has elevated JE in the public health agenda of preventable childhood diseases, and surveillance of acute neurological infections to confirm polio eradication, simultaneously, has led to a greater awareness of the disease burden attributable to JE. The only internationally licensed JE vaccine, an inactivated mouse-brain derived vaccine, is efficacious but is problematic from the perspectives of reactogenicity, requirement for numerous doses, cost and reliance on a neurological tissue substrate. A live-attenuated vaccine distributed only in China also is efficacious and requires fewer doses; however, production and regulatory standards are unresolved. Several approaches toward developing novel JE vaccines that could fill the gap in JE vaccine need are under pursuit. The minutes and recommendations of a meeting of experts to discuss these issues, jointly sponsored by the World Health Organization and the Children's Vaccine Initiative in Bangkok, Thailand, 13-15 October, 1998, are presented. PMID- 10821970 TI - Points to consider in the development of a surrogate for efficacy of novel Japanese encephalitis virus vaccines. AB - Although an effective killed virus vaccine to prevent illness due to Japanese encephalitis virus (JEV) infection exists, many authorities recognize that a safe, effective live JEV vaccine is desirable in order to reduce the cost and the number of doses of vaccine required per immunization. A large-scale clinical efficacy trail for such a vaccine would be both unethical and impractical. Therefore, a surrogate for the efficacy of JE vaccines should be established. Detection of virus-neutralizing antibodies in sera of vaccinees could constitute such a surrogate for efficacy. Field studies of vaccinees in endemic areas and studies done in mice already exist to support this concept. Also, titers of virus neutralizing antibodies are already accepted as a surrogate for the efficacy of yellow fever virus vaccines and for the efficacy of other viral vaccines as well. In developing a correlation between N antibody titers and protection from JEV infection, standard procedures must be validated and adopted for both measuring N antibodies and for testing in animals. A novel live virus vaccine could be tested in the mouse and/or the monkey model of JEV infection to establish a correlation between virus-neutralizing antibodies elicited by the vaccines and protection from encephalitis. In addition, sera of subjects receiving the novel live JEV vaccine in early clinical trials could be passively transferred to mice or monkeys in order to establish the protective immunogenicity of the vaccine in humans. A monkey model for JEV infection was recently established by scientists at WRAIR in the US. From this group, pools of JEV of known infectivity for Rhesus macaques may be obtained for testing of immunity elicited by live JE vaccine virus. PMID- 10821971 TI - Immunogenicity and protective efficacy of the current inactivated Japanese encephalitis vaccine against different Japanese encephalitis virus strains. AB - Mouse brain-derived, inactivated Japanese encephalitis (JE) vaccine has been internationally used for many years. It is believed that this vaccine made a great contribution to the reduction of JE patients in several countries. Mouse brain-derived, Beijing-1 and Nakayama JE vaccines induce high levels of neutralizing antibodies. High levels of induced antibodies are maintained at least for 3-4 yr. The induced antibodies are cross-reactive to heterologous strains; however, the neutralizing antibody titers against heterologous strains are usually lower than those against homologous strains. Considering that both Nakayama and Beijing-1 JE vaccines showed high levels of protective efficacy in Taiwan and Thailand where strains other than Nakayama and Beijing-1 were circulating, we conclude that the current inactivated JE vaccine can induce high levels of protective immunity against heterologous JE virus strains. PMID- 10821972 TI - Phylogenetic analysis suggests only one serotype of Japanese encephalitis virus. AB - Phylogenetic analysis was performed for different genome regions of Japanese encephalitis virus (JEV). Similar genetic groupings were identified for all analyzed genome regions including complete genomes. More extensive analysis was performed for 92 isolates (complete envelope sequences) available in the GenBank. Results of phylogenetic analysis were compared with those performed for human positive strand RNA viruses with well characterized serotypes - poliovirus (PV) and dengue virus (DEN). The observed level of the JEV inter-genotype diversity was much less than that observed across PV and DEN serotypes and was consistent with the genetic diversity observed within PV or DEN serotypes. This genetic analysis supports the contention that all known JEV isolates comprise a single serotype. PMID- 10821973 TI - Live attenuated tetravalent dengue vaccine. AB - The development of a live attenuated tetravalent dengue vaccine is currently the best strategy to obtain a vaccine against dengue viruses. The Mahidol University group developed candidate live attenuated vaccines by attenuation through serial passages in certified primary cell cultures. Dengue serotype 1, 2 and 4 viruses were developed in primary dog kidney cells, whereas dengue serotype 3 was serially passaged in primary African green monkey kidney cells. Tissue culture passaged strain viruses were subjected to biological marker studies. Candidate vaccines have been tested as monovalent (single virus), bivalent (two viruses), trivalent (three viruses) and tetravalent (all four serotype viruses) vaccines in Thai volunteers. They were found to be safe and immunogenic in both adults and children. The Mahidol live attenuated dengue 2 virus was also tested in American volunteers and resulted in good immune response indistinguishable from those induced in Thai volunteers. The master seeds from the four live attenuated virus strains developed were provided to Pasteur Merieux Connaught of France for production on an industrial scale following good manufacturing practice guidelines. PMID- 10821974 TI - Liver cirrhosis and peptic ulcer disease; a correlation with Helicobacter pylori? PMID- 10821975 TI - Helicobacter pylori seroprevalence in cirrhotic patients with hepatitis B virus infection. AB - Liver cirrhosis is a significant cause of death in Italy and one of the most frequent causes of hospitalization. The burden of cirrhotic patients on the National Health System is extremely high due to the frequent need for medical care. Acute peptic ulcer and upper gastrointestinal bleeding reportedly occur in over one-third of cirrhotic patients. Since Helicobacter pylori (H. pylori) infection strongly correlates with peptic ulcer, we wished to ascertain the prevalence of H. pylori infection in cirrhotic patients. In a case-control study we looked for this infection in 45 consecutive male patients suffering from hepatitis B virus (HBV)-related cirrhosis and 310 sex and age matched blood donors resident in the same area. Antibodies against H. pylori were present in 40/45 (89%) patients and 183/310 (59%) blood donors (P<0.001). This very high prevalence of H. pylori may explain the frequent occurrence of gastroduodenal ulcer in cirrhotic patients. (See Editorial p. 203) PMID- 10821976 TI - Incidence and onset of critical illness polyneuropathy in patients with septic shock. AB - BACKGROUND: To estimate the incidence and onset of critical illness polyneuropathy (CIP) in patients in septic shock. METHODS: Prospective, observational study, no interventions, in a general 9-bed ICU of a large teaching hospital. Twenty-five patients consecutively admitted to the ICU for treatment of septic shock were studied. Within 72 h of admission to the ICU a complete neurological examination and electromyografic studies were done. Studies were repeated weekly until discharge of ICU or death or CIP confirmed. RESULTS: Nineteen patients developed CIP (76%), with a majority (80%) within 72 h after onset of septic shock. All twenty-two patients with multi organ dysfunction syndrome (MODS) had CIP. The three patients without MODS did not have CIP (P<0.01). CONCLUSIONS: In a group of patients suffering from septic shock the incidence of CIP is high (76%). The onset is early, within 72 h after onset of septic shock. CIP is an early feature of MODS, developing after septic shock. PMID- 10821977 TI - Hypercapnia in obstructive sleep apnoea syndrome. AB - BACKGROUND: The reports on the prevalence of hypercapnia in Obstructive Sleep Apnoea Syndrome (OSAS) are conflicting. We studied the prevalence of hypercapnia in a population of OSAS patients referred to a Department of Respiratory Medicine and the mechanism of the respiratory failure in OSAS associated with Obesity Hypoventilation Syndrome (OHS) or with Chronic Obstructive Pulmonary Disease (COPD) (Overlap syndrome). METHODS: We studied 219 consecutive OSAS patients during a period of 3 years. We recorded age and anthropomorphic data and performed polysomnography and pulmonary function tests. In relation to the value of PaCO(2), the patients were divided in hypercapnic (PaCO(2)>45 mmHg) patients and normocapnic patients. They were also divided into three groups in relation to the presence of "simple" or "pure" OSAS, to the presence of OSAS associated with COPD, to the presence of OSAS associated with OHS. RESULTS: Seventeen per cent of the patients were hypercapnic. They were significantly heavier, had more severe lung function test abnormalities and more severe nocturnal oxyhemoglobin desaturations than the normocapnic ones, while Forced Expiratory Volume in one second as a percentage of Forced Vital Capacity (FEV1/FVC %) and Apnoea/Hypopnoea Index (AHI) were similar. OHS patients (13%) were significantly younger and heavier, had lower PaO(2) and higher PaCO(2) than "simple" OSAS patients (77%) and Overlap patients (10%) and had more severe restrictive defect. There was no difference in terms of AHI among the three groups, but nocturnal oxyhemoglobin desaturations were more severe in OHS group. In OHS group hypercapnia was correlated to FVC% of predicted and FEV1% of predicted and to the mean nocturnal oxyhemoglobin saturation; in Overlap patients PaCO(2) was correlated to Forced Expiratory Flow rate at low Vital Capacity. CONCLUSION: Seventeen per cent of OSAS patients referred to a Department of Respiratory Medicine were hypercapnic. Hypercapnia in OHS patients correlates to the restrictive ventilatory defect whereas in Overlap patients it seems to correlate to peripheral airways obstruction. The distinction between patients with "simple" or "pure" OSAS and patients affected by OSAS associated with OHS or COPD could be important not only for clinical and prognostic implications, but also for the consequences in terms of ventilatory treatment. PMID- 10821978 TI - A bone biopsy is mandatory in the optimal management of bone lesions in patients with a long-term history of malignancy of the breast. AB - Bone lesions, present in two women with primary breast cancer treated more than 5 years ago, were investigated thoroughly by biochemical markers and imaging techniques. The definite diagnosis was established by bone biopsy, which revealed two totally unrelated disorders that altered the management and prognosis for these two patients. One patient had Paget's disease, and the other had metastatic hepatocellular carcinoma. PMID- 10821979 TI - Acquired hemophilia. PMID- 10821980 TI - Severe acute lung injury induced by gemcitabine. AB - Gemcitabine is a nucleoside analog that is active in the treatment of various solid tumors. In general it is well tolerated and has few side effects. Pulmonary toxicity reported with gemcitabine use is usually mild and self-limiting. We present a case of severe pulmonary dysfunction after intravenous administration of a single dose of gemcitabine in a 58-year-old female patient with metastatic carcinoma of the pancreas. She developed tachypnea, marked hypoxemia, and an interstitial infiltrate on chest radiograph consistent with pulmonary edema, 4 days after receiving this drug. Diuretics and corticosteroids were beneficial in treating the acute respiratory failure. Pulmonary damage was completely resolved by means of clinical and radiological assessment. Because of the severity of this side effect, no further treatment with gemcitabine was given. Eventually, the patient died because of obstruction of the bowel due to progression of tumor growth. Publications concerning severe pulmonary toxicity due to gemcitabine are sparse. Pathophysiology and treatment are considered and a review of the literature is presented. PMID- 10821981 TI - Protein synthesis in axons and terminals: significance for maintenance, plasticity and regulation of phenotype. With a critique of slow transport theory. AB - This article focuses on local protein synthesis as a basis for maintaining axoplasmic mass, and expression of plasticity in axons and terminals. Recent evidence of discrete ribosomal domains, subjacent to the axolemma, which are distributed at intermittent intervals along axons, are described. Studies of locally synthesized proteins, and proteins encoded by RNA transcripts in axons indicate that the latter comprise constituents of the so-called slow transport rate groups. A comprehensive review and analysis of published data on synaptosomes and identified presynaptic terminals warrants the conclusion that a cytoribosomal machinery is present, and that protein synthesis could play a role in long-term changes of modifiable synapses. The concept that all axonal proteins are supplied by slow transport after synthesis in the perikaryon is challenged because the underlying assumptions of the model are discordant with known metabolic principles. The flawed slow transport model is supplanted by a metabolic model that is supported by evidence of local synthesis and turnover of proteins in axons. A comparison of the relative strengths of the two models shows that, unlike the local synthesis model, the slow transport model fails as a credible theoretical construct to account for axons and terminals as we know them. Evidence for a dynamic anatomy of axons is presented. It is proposed that a distributed "sprouting program," which governs local plasticity of axons, is regulated by environmental cues, and ultimately depends on local synthesis. In this respect, nerve regeneration is treated as a special case of the sprouting program. The term merotrophism is proposed to denote a class of phenomena, in which regional phenotype changes are regulated locally without specific involvement of the neuronal nucleus. PMID- 10821982 TI - Functional changes of the basal ganglia circuitry in Parkinson's disease. AB - The basal ganglia circuitry processes the signals that flow from the cortex, allowing the correct execution of voluntary movements. In Parkinson's disease, the degeneration of dopaminergic neurons of the substantia nigra pars compacta triggers a cascade of functional changes affecting the whole basal ganglia network. The most relevant alterations affect the output nuclei of the circuit, the medial globus pallidus and substantia nigra pars reticulata, which become hyperactive. Such hyperactivity is sustained by the enhanced glutamatergic inputs that the output nuclei receive from the subthalamic nucleus. The mechanisms leading to the subthalamic disinhibition are still poorly understood. According to the current model of basal ganglia organization, the phenomenon is due to a decrease in the inhibitory control exerted over the subthalamic nucleus by the lateral globus pallidus. Recent data, however, suggest that additional if not alternative mechanisms may underlie subthalamic hyperactivity. In particular, given the reciprocal innervation of the substantia nigra pars compacta and the subthalamic nucleus, the dopaminergic deficit might influence the subthalamic activity, directly. In addition, the increased excitatory drive to the dopaminergic nigral neurons originating from the hyperactive subthalamic nucleus might sustain the progression of the degenerative process. The identification of the role of the subthalamic nucleus and, more in general, of the glutamatergic mechanisms in the pathophysiology of Parkinson's disease might lead to a new approach in the pharmacological treatment of the disease. Current therapeutic strategies rely on the use of L-DOPA and/or dopamine agonists to correct the dopaminergic deficit. Drugs capable of antagonizing the effects of glutamate might represent, in the next future, a valuable tool for the development of new symptomatic and neuroprotective strategies for therapy of Parkinson's disease. PMID- 10821983 TI - Metamorphosis in drosophila and other insects: the fate of neurons throughout the stages. AB - The nervous system of insects is profoundly reorganised during metamorphosis, affecting the fate of different types of neuron in different ways. Almost all adult motor neurons derive from larval motor neurons that are respecified for adult functions. A subset of larval motor neurons, those which mediate larval- or ecdysis-specific behaviours, die before and immediately after eclosion, respectively. Many adult interneurons develop from larval interneurons, whereas those related to complex adult sense organs originate during larval life from persisting embryonic neuroblasts. Sensory neurons of larvae and adults derive from essentially two distinct sources. Larval sensory neurons are formed in the embryonic integument and - with few exceptions - die during metamorphosis. Their adult counterparts, on the other hand, arise from imaginal discs. Special emphasis is given in this review to the metamorphic remodelling of persisting neurons, both at the input and output levels, and to the associated behavioural changes. Other sections deal with the programmed death of motor neurons and its causes, as well as with the metamorphic interactions between motor neurons and their target muscles. Remodelling and apoptosis of these two elements appear to be under independent ecdysteroid control. This review focusses on the two most thoroughly studied holometabolous species, the fruitfly Drosophila melanogaster and the tobacco hornworm moth Manduca sexta. While Manduca has a long tradition in neurodevelopmental studies due to the identification of many of its neurons, Drosophila has been increasingly used to investigate neural reorganisation thanks to neurogenetic tools and molecular approaches. The wealth of information available emphasises the strength of the insect model system used in developmental studies, rendering it clearly the most important system for studies at the cellular level. PMID- 10821984 TI - Parent training skills and methadone maintenance: clinical opportunities and challenges. AB - Children raised in substance abusing families show high rates of behavioural and emotional problems, in particular oppositional, defiant and non-compliant behaviours. While a range of social and individual factors correlate with poor parenting, it is often the quality of the parent-child relationship that mediates the effects of most other risk factors on child development. By addressing this relationship using behavioural family interventions, child behaviour problems have been reduced in multiple problem families. However, there has been little attempt to systematically evaluate such programs in substance abusing families. It is argued that methadone replacement programs provide a window of opportunity to deliver well-validated parent training programs that enhance the quality of parent-child relations. However, it is likely that such programs would need to be medium to long term and address issues beyond parent child relationships. How such interventions may be delivered and evaluated is discussed. PMID- 10821985 TI - Clinical pharmacology of oral cotinine. AB - Cotinine is the major proximate metabolite of nicotine. The aims of our study were to assess the pharmacokinetics of oral cotinine comparing the use of saliva and plasma concentrations, and to characterize the subjective and cardiovascular effects of oral cotinine in nonsmokers. The clearance and half-life of cotinine measured using plasma or saliva concentrations were similar. There was no change in heart rate or blood pressure, and no differences in subjective response with cotinine compared to placebo. We conclude that administration of oral cotinine with measurement in saliva samples is easy, safe, and provides an accurate estimate of systemic clearance and half-life of cotinine. PMID- 10821986 TI - Self-report stability of adolescent substance use: are there differences for gender, ethnicity and age? AB - This study used the National Longitudinal Survey of Youth and is the first to examine a 2-year report stability of substance use among adolescents while stratifying for gender, ethnicity, and age. This study examined lifetime use and age at onset report stability, and the internal consistency of reports while excluding nonusers and incident cases (respondents who may have initiated substance use between the two reporting periods) from the analyses. Report agreement of lifetime use for each substance was over 80% and was highest among alcohol users and lowest for cigarette and marijuana users. Report agreement was higher for female compared to male cigarette users. External consistency of lifetime use of cigarettes and marijuana was higher for whites compared to Hispanic or African American adolescents. Internal consistency was high but lifetime use reports were more stable than age at onset reports. PMID- 10821987 TI - Nalmefene: blockade of intravenous morphine challenge effects in opioid abusing humans. AB - No studies have assessed the dose-effect or duration of opioid blockade in opioid abusers produced by oral nalmefene, a micro-opioid antagonist. The present study examined the profile and time course of oral nalmefene blockade of subjective and physiological effects produced by intravenous morphine. To assess these effects, seven opioid abusers received oral nalmefene (0, 50 and 100 mg) followed by intravenous morphine (0, 10 and 20 mg) challenges every 24 h for 96 h using a Latin square randomized cross-over design. The duration of blockade varied by measure and dose. Both 50 and 100 mg nalmefene blocked morphine's effects up to 48 h. PMID- 10821988 TI - Buprenorphine: a controlled clinical trial in the treatment of opioid dependence. AB - Clinical trials carried out to compare methadone and buprenorphine in the treatment of opioid dependence have generally employed an alcoholic solution of buprenorphine, which has a bioavailability superior to that of the tablets. Since the product available for large scale use is in tablet form, one intended to verify the efficacy of this formulation. In a multicentre randomised controlled double blind study, 72 opioid dependent patients were assigned to treatment with buprenorphine (8 mg/day) or methadone (60 mg/day) for a period of 6 months. The two compounds did not show any significant difference with regard to urinalyses: the average percentage of analyses proving negative was 60.4% for patients assigned to buprenorphine, and 65.5% for those assigned to methadone. With regard to retention, a non-significant trend in favour of methadone was observed. Patients completing the trial improved significantly in terms of psychosocial adjustment and global functioning, as ascertained by the DSM-IV-GAF and symptom checklist-90 (SCL-90) scales, and this was independent of the treatment group. Finally, in the case of buprenorphine, patients who dropped out differed significantly from those who stayed, in terms of a higher level of psychopathological symptoms, and a lower level of psychosocial functioning. The results of the study further support the utility of buprenorphine for the treatment of opioid dependence. PMID- 10821989 TI - Use of buprenorphine in HIV-infected injection drug users: negligible impact on virologic response to HAART. The Manif-2000 Study Group. AB - Some HIV-infected injecting drug users (IDUs) on drug abuse maintenance treatment have access to highly active antiretroviral therapy (HAART); this raises questions about the effects of individual treatments on the efficacy of HAART. The French Cohort Study of HIV-infected IDUs - MANIF-2000 - allowed one to assess whether buprenorphine differentially impacts efficacy of HAART. Of the 103 HAART treated patients, (excluding active IDUs and patients on methadone), 20 were on buprenorphine substitution treatment and 83 were ex-IDUs. A linear regression model used the differences in viral load titre before and after treatment initiation, as a dependent variable, and showed that buprenorphine treatment was not significantly associated with viral load trend. This was also the case when adjusting for other potential confounders, and suggests that there is no major short-term influence of buprenorphine on HIV viral load in HAART-treated patients. PMID- 10821990 TI - Initiating abstinence in cocaine abusing dually diagnosed homeless persons. AB - This study measured effectiveness of behavioral day treatment plus abstinence contingent housing and work therapy (DT+) versus behavioral day treatment alone (DT). A randomized controlled trial assessed participants at baseline, 2 and 6 months. Participants (N=110) met criteria for cocaine abuse or dependence, non psychotic mental disorders, and homelessness. DT+ achieved greater abstinence at 2 and 6 months and more days housed at 6 months than DT. Effectiveness of DT+ was demonstrated, with greatest impacts on abstinence outcomes. Results replicated earlier work demonstrating effectiveness of behavioral day treatment and contingency management as an effective combination for cocaine abusing homeless persons. PMID- 10821991 TI - Carvedilol affects the physiological and behavioral response to smoked cocaine in humans. AB - The noradrenergic system is implicated in mediating some of the physiological effects of cocaine. The purpose of this study was to investigate whether treatment with an adrenergic blocker, carvedilol, which would be expected to attenuate the physiological effects of cocaine, would also attenuate the subjective and behavioral response to cocaine in humans. Twelve crack cocaine users participated in this double-blind, placebo-controlled outpatient study. Acute treatment with 50 mg of oral carvedilol attenuated the smoked cocaine induced increases in heart rate, systolic and diastolic blood pressure. The number of cocaine self-administrations was lower under 25 mg carvedilol treatment condition compared with 50 mg carvedilol or placebo treatment conditions. The subjective responses to smoked cocaine deliveries were not affected by carvedilol treatment. These results suggest that acute treatment with carvedilol attenuates the physiological effects of smoked cocaine. The effects of carvedilol on cocaine self-administration need to be studied further. PMID- 10821992 TI - Drug abuse, methadone treatment, and health services use among injection drug users with AIDS. AB - This paper compares health care use across subgroups of injection drug users (IDUs) with AIDS, as defined by current drug abuse status and participation in methadone maintenance treatment (MMT), using surveillance-identified IDU status and health care claims data. Merged Medicaid and AIDS surveillance data were analyzed using ordinary least squares regression, simple logistic regression and multinomial logistic regression. Consistent MMT was more likely among women, Whites and older subjects. Monthly total expenditures and inpatient expenditures were significantly lower for IDUs in MMT than for IDUs with claims indicative of current drug abuse. Consistent participation in MMT was associated with a higher probability of antiretroviral use and, among antiretroviral users, more consistent use of antiretrovirals. Merged administrative data sets can be an important data source that illuminate the relationships among drug abuse, drug treatment, and HIV-related health care. For AIDS-infected IDUs, consistent MMT may lower barriers to receipt of appropriate HIV-related health care and reinforce adherence to medical recommendations. PMID- 10821993 TI - Anabolic-androgenic steroid use disorders among a sample of Australian competitive and recreational users. AB - Illicit anabolic-androgenic steroid (AAS) use, for body building and body image purposes, is increasing internationally. This study describes the prevalence and symptoms of AAS use disorders obtained using a semi-structured interview among a mixed-gender sample of 100 current Australian users, that were recruited from a variety of sources. The median age of the sample was 27 years and 94% were male. The full range of DSM IV symptoms of AAS abuse and dependence were reported and over three quarters (78%) of the sample exhibited at least one symptom of abuse or dependence on AAS. A total of 23% of the participants qualified for a diagnosis of AAS dependence using DSM IV criteria and a further 25% met criteria for AAS abuse. There were no gender differences in AAS abuse or dependence diagnoses. The only variable related to an AAS substance use disorder was reporting the experience of AAS-related aggression, which may be a useful clinical indicator of the disorder. PMID- 10821994 TI - Incorporating the AUDIT into a general population telephone survey: a methodological experiment. AB - This study assessed potential ordering and wording effects of the alcohol use disorders identification test (AUDIT). In total, 688 respondents were randomly assigned to one of four experimental conditions: Intact order/original wording (n=148), intact order/revised wording (n=183), split order/original wording (n=192), split order/revised wording (n=166). Changes to question order and wording had no discernable impact on the scores of the AUDIT. Our results suggest that alterations to the AUDIT can be made in order to integrate it within a larger survey without adversely affecting its measurement properties. PMID- 10821995 TI - Psychobiological problems in heavy 'ecstasy' (MDMA) polydrug users. AB - Twelve heavy recreational ecstasy drug users (30-1000 occasions), 16 light ecstasy users (1-20 occasions) and 22 non ecstasy user controls, with group mean ages around 21 years, were compared. Three self-rating questionnaires were completed when drug-free: the SCL-90 (an outpatient psychiatric symptom checklist), the impulsiveness venturesomeness and empathy (IVE) scale; and the uplifts, hassles, stresses and cognitive failures questionnaire. Heavy Ecstasy users reported significantly higher scores than controls on the following SCL-90 factors: paranoid ideation, psychoticism, somatisation, obsessionality, anxiety, hostility, phobic anxiety, altered appetite and restless sleep, together with greater IVE impulsiveness. Light ecstasy users generally produced intermediate scores, with significantly higher scores than controls on two factors and significantly lower scores than heavy ecstasy users on another two. Previous reports have described various psychiatric and psychobiological disorders in recreational ecstasy users, but it is not known how typical they are, being mainly based on individual case studies. This is the first study to describe psychological problems in a non clinical sample of young recreational ecstasy users. However, our ecstasy users were polydrug users, with both groups showing significantly greater usage of amphetamine, LSD and cocaine, than the controls. These other illicit drugs probably contributed to their adverse psychobiological profiles, while there is also the possibility of pre-existing differences between ecstasy users and non users. However, since repeated MDMA can cause serotonergic neurotoxicity in laboratory animals and man, these problems may reflect reduced serotonin activity induced by regular ecstasy use. PMID- 10821996 TI - Increased left posterior parietal-temporal cortex activation after D-fenfluramine in women with panic disorder. AB - It is unclear whether the functional changes found in panic disorder reflect disturbed physiology of particular neurotransmitters. One method of investigating altered neurotransmission is to assess regional brain activations in response to agonist challenges. D-Fenfluramine is a medication that induces neuronal release of serotonin. Using ?15OH(2)O and positron emission tomography (PET), measurements of regional cerebral blood flow (rCBF) were done at t=-20, -5, +20 and +35 relative to the IV D-fenfluramine injection (t=0) in nine panic disordered and 18 healthy subjects. Subjects were otherwise healthy, right handed, non-smoking and not taking psychotropic medication. ?15OH(2)O PET scans were assessed with Statistical Parametric Mapping using individual global cerebral blood flow as a covariate. Comparisons of the (baseline) first two scans between healthy and panic-disordered subjects showed a decreased rCBF in the left posterior parietal-superior temporal cortex in the patient group. Fenfluramine induced increases as defined by the last two scans minus the first two scans were compared between groups and a significantly greater increase in the same left posterior parietal-superior temporal region was found in panic-disordered subjects. Consistent with this finding, differences between the last two scans (post-fenfluramine) of the healthy and panic-disordered subjects showed an increased rCBF in the left superior temporal cortex in panic-disordered subjects. Functional pathology in the left parietal-superior temporal cortex in panic disorder may be related to abnormal modulation by serotonin. PMID- 10821997 TI - Cortical response to motor stimulation in neuroleptic-naive first episode schizophrenics. AB - The purpose of the present study was to evaluate the cortical response to motor stimulation in neuroleptic-naive first episode schizophrenics in comparison to matched controls using a high speed functional magnetic resonance imaging technique (fMRI). Twelve patients satisfying ICD 10 criteria (F20.0) for schizophrenia (paranoid subtype) as well as sex- and age-matched healthy volunteers participated in this study. All subjects underwent fMRI examination on a conventional 1.5 T MR unit equipped with an echo-planar imaging booster. The blood oxygen level dependent (BOLD) response of the sensorimotor cortex and the higher order SMA region was evaluated during performance of a left hand sequential finger opposition task. Special care was taken to minimize performance and motion artifacts. Patients and controls showed no notable difference with respect to laterality, changes of signal intensity or spatial extent of activation within the primary and higher order motor regions. Using high speed fMRI no fundamental motor cortical dysfunction was evident in a group of paranoid neuroleptic-naive first episode schizophrenic patients. In contrast to data previously reported for chronic disorganized medicated patients, these results suggest that motor dysfunction is not part of the phenomenology of acute paranoid first episode patients. PMID- 10821998 TI - Thalamic abnormalities in patients with schizophrenia revealed by proton magnetic resonance spectroscopy. AB - Recent investigations suggest that thalamic abnormalities may underlie symptom formation in schizophrenia. We previously demonstrated reduced concentrations of N-acetylaspartate (NAA) in tissue from the thalamus of schizophrenic patients using in vitro proton magnetic resonance spectroscopy (1H-MRS). In the present study, in vivo 1H-MR spectra of the left thalamus and frontal lobe were investigated in 20 patients with schizophrenia and 16 age-matched control subjects to replicate our previous postmortem findings and support the hypothesis of thalamic abnormality in schizophrenia. Schizophrenic patients showed significantly lower NAA/total creatine (Cr) and choline-containing compounds (Cho)/Cr ratios in the thalamus than control subjects, while no significant difference was found in the frontal lobe. There was no significant correlation in the schizophrenic patients between the NAA/Cr or Cho/Cr ratio and other clinical data including clinical symptoms or neuroleptic dosage. These findings may further support other studies suggesting decreased thalamic volume or neuronal number and/or thalamic dysfunction, and reduction in size of white matter tracts adjacent to the thalamus in schizophrenia, as well as our previous postmortem MRS study. PMID- 10821999 TI - Hippocampal pathology in schizophrenia: magnetic resonance imaging and spectroscopy studies. AB - The hippocampus is a site of previously reported structural and functional abnormalities in schizophrenia. We used magnetic resonance imaging (MRI) and proton magnetic resonance spectroscopy (MRS) to measure gray matter volumes, the neuronal marker N-acetylaspartate (NAA), and the combination of glutamate (Glu), glutamine (Gln), and gamma-aminobutyric acid (GABA), designated Glx. Measurements were obtained of the medial temporal lobe, centered on the hippocampus, in 10 male patients with schizophrenia (3 neuroleptic-medicated and 7 medication-free), and 10 matched normal volunteers. MRI volumetric measurements and MRS data obtained with short echo time (TE=20 ms) one-dimensional STEAM chemical shift imaging (CSI) on a GE 1.5 Tesla Signa system were analyzed. A laterality index ?(L-R)/(L+R) was generated from the ratio of Glx to choline-containing compounds (Cho) to test asymmetry changes. Reliability of the MRS measures was assessed with five test-retest studies of healthy volunteers and showed coefficients of variation (CV) in the range of 36-44% for the MRS ratios and standard deviations (S.D.) of 0.15-0.17 for the laterality indices. The Glx/Cho laterality index showed a relative right-sided excess in this region in the patients (-0.23+/ 0.20) compared to the controls (+0.06+/-0.20), which was not confounded by tissue composition or placement variability of the MRS voxels. Hippocampal volume deficit and asymmetry were not significant, and other MRS measures showed no differences between patients and controls. The preliminary finding of a lateralized abnormality in Glx is consistent with postmortem findings of asymmetric neurochemical temporal lobe abnormalities in schizophrenia. PMID- 10822000 TI - An estimation of the first positive Lyapunov exponent of the EEG in patients with schizophrenia. AB - We studied the complexity of the electroencephalogram (EEG) in schizophrenic patients by estimating the first Lyapunov exponent (L1), which might serve as an indicator of the specific brain function in schizophrenia. We recorded the EEG from 25 schizophrenic patients (12 male, 13 female; age=25.1+/-7.0 years) fulfilling DSM-IV criteria and 15 healthy controls (9 male, 6 female; age=27. 8+/ 4.2 years) at 16 electrodes, different from previous studies which recorded the EEGs at limited electrodes. We employed a method with an optimal embedding dimension to calculate the L1s. For limited noisy data, this algorithm was strikingly faster and more accurate than previous ones. Our results showed that the schizophrenic patients had lower values of the L1 at the left inferior frontal and anterior temporal regions compared with normal controls. These results for L1 in non-linear analysis have some differences from those for power ratios in linear analysis. These suggest that the non-linear analysis of the EEGs such as the estimation of the L1 might be a useful tool in analyzing EEG data to explore the neurodynamics of the brains of schizophrenic patients. PMID- 10822008 TI - Cholesterol oxidase: sources, physical properties and analytical applications. AB - Since Flegg (H.M. Flegg, An investigation of the determination of serum cholesterol by an enzymatic method, Ann. Clin. Biochem. 10 (1973) 79-84) and Richmond (W. Richmond, The development of an enzymatic technique for the assay of cholesterol in biological fluids, Scand. J. clin. Lab. Invest. 29 (1972) 25; W. Richmond, Preparation and properties of a bacterial cholesterol oxidase from Nocardia sp. and its application to enzyme assay of total cholesterol in serum, Clinical Chemistry 19 (1973) 1350-1356) first illustrated the suitability of cholesterol oxidase (COD) for the analysis of serum cholesterol, COD has risen to become the most widely used enzyme in clinical laboratories with the exception of glucose oxidase (GOD). The use is widespread because assays incorporating the enzyme are extremely simple, specific, and highly sensitive and thus offer distinct advantages over the Liebermann-Burchard analytical methodologies which employ corrosive reagents and can be prone to unreliable results due to interfering substances such as bilirubin. Individuals can now readily determine their own serum cholesterol levels with a simple disposable test kit. This review discusses COD in some detail and includes the topics: (1) The variety of bacterial sources available; (2) The various extraction/purification protocols utilised in order to obtain protein of sufficient clarification (purity) for use in food/clinical analysis; (3) Significant differences in the properties of the individual enzymes; (4) Substrate specificities of the various enzymes; (5) Examples of biological assays which have employed cholesterol oxidase as an integral part of the analysis, and the various assay protocols; (6) New steroidal products of COD. This review is not a comprehensive description of published work, but is intended to provide an account of recent and current research, and should promote further interest in the application of enzymes to analytical selectivity. PMID- 10822009 TI - Estrogen and aryl hydrocarbon receptor expression and crosstalk in human Ishikawa endometrial cancer cells. AB - Ishikawa endometrial cancer cells express the estrogen receptor (ER), and this study investigates aryl hydrocarbon receptor (AhR) expression and inhibitory AhR ER crosstalk in this cell line. Treatment of Ishikawa cells with the AhR agonist [3H]2,3,7, 8-tetrachlorodibenzo-p-dioxin (TCDD) gave a radiolabeled nuclear complex that sedimented at 6.0 S in sucrose density gradients, and Western blot analysis confirmed that Ishikawa cells expressed human AhR and AhR nuclear translocator (Arnt) proteins. Treatment of Ishikawa cells with 10 nM TCDD induced a 9.7-fold increase in CYP1A1-dependent ethoxyresorufin O-deethylase (EROD) activity and a 10.5-fold increase in chloramphenicol acetyltransferase (CAT) activity in cells transfected with pRNH11c containing an Ah-responsive human CYP1A1 gene promoter insert (-1142 to +2434). Inhibitory AhR-ER crosstalk was investigated in Ishikawa cells using E2-induced cell proliferation and transcriptional activation assays in cells transfected with E2-responsive constructs containing promoter inserts from the progesterone receptor and vitellogenin A2 genes. AhR agonists including TCDD, benzo[a]pyrene (BaP) and 6 methyl-1,3,8-trichlorodibenzofuran, inhibited 32-47% of the E2-induced responses. In contrast, neither estrogen nor progesterone inhibited EROD activity induced by TCDD in Ishikawa cells, whereas inhibitory ER-AhR crosstalk was observed in ECC-1 endometrial cells suggesting that these interactions were cell context-dependent. PMID- 10822010 TI - Partial purification and biochemical characterization of a membrane glucocorticoid receptor from an amphibian brain. AB - A membrane receptor for corticosterone (mGR) in the brain of the roughskin newt (Taricha granulosa) has been previously identified. This manuscript reports the evaluation of several chromatographic resins for enrichment of the newt mGR solubilized from neuronal membranes. A protein with an apparent molecular weight of 63 kDa was purified to near homogeneity following sequential purification using ammonium sulfate fractionation, wheat germ agglutinin (WGA)-agarose chromatography, hydroxylapatite chromatography, and an immobilized ligand affinity resin (Corticosterone-Sepharose). Other studies employed a novel protein differential display strategy and a photoaffinity labeling strategy to visualize candidate receptor proteins following SDS-PAGE. Both of these techniques also identified a 63 kDa protein, agreeing with the estimation of molecular weight from the purification data. Furthermore, the use of 2D SDS-PAGE following the photolabeling procedure showed the candidate 63 kDa protein to have a pI of approximately 5.0. Taken together these data suggest that the newt mGR is an acidic glycoprotein with an apparent molecular weight of 63 kDa. Because these characteristics of newt mGR are inconsistent with the characteristics of intracellular glucocorticoid receptors, these two receptor proteins are apparently distinct. PMID- 10822011 TI - Control of estrogen receptor ligand binding by Hsp90. AB - The molecular chaperone Hsp90 interacts with unliganded steroid hormone receptors and regulates their activity. We have analyzed the function of yeast and mammalian Hsp90 in regulating the ability of the human estrogen receptor (ER) to bind ligands in vivo and in vitro. Using the yeast system, we show that the ER expressed in several different hsp82 mutant strains binds reduced amounts of the synthetic estrogen diethylstilbestrol compared to the wild type. This defect in hormone binding occurs without any significant change in the steady state levels of ER protein. To analyze the role of mammalian Hsp90, we synthesized the human ER in rabbit reticulocyte lysates containing geldanamycin, an Hsp90 inhibitor. At low concentrations of geldanamycin we observed reduced levels of hormone binding by the ER. At higher concentrations, we found reduced synthesis of the receptor. These data indicate that Hsp90 functions to maintain the ER in a high affinity hormone-binding conformation. PMID- 10822012 TI - Cloning and expression of the bovine 11beta-hydroxysteroid dehydrogenase type-2. AB - The bovine 11beta-hydroxysteroid dehydrogenase type 2 enzyme (11beta-HSD-2) cDNA was cloned from three overlapping PCR fragments using primers based on the human and ovine 11beta-HSD-2 cDNA sequences. Both cDNA ends were obtained by a modified RACE (Rapid Amplification of cDNA Ends) method. The bovine 11beta-HSD-2 cDNA is 1878 bp long, excluding the poly(A) tail. It consists of a 5'-untranslated region of 133 bp, an open reading frame of 1215 bp and a 3'-untranslated region of 530 bp. Bovine 11beta-HSD-2 cDNA is highly homologous to that of the sheep (92%) and less related to the human (67%), rabbit (65%), rat (52%) and mouse (45%) cDNA. The predicted bovine 11beta-HSD-2 protein contains 404 amino acid residues with a calculated mol wt of 43,985. It is homologous to the sheep (98%) and human (88%) protein, and less related to that of the rabbit (76%), rat (80%) and mouse (77%). The cloned 11beta-HSD-2 cDNA was transfected into CHOP cells and the enzymatic characteristics determined. The enzyme functions primarily as an oxidase, uses NAD(+) and is more active with corticosterone as a substrate than with cortisol or dexamethasone. It is expressed in high concentrations in kidney, adrenal and colon, and in small concentrations in liver, heart and lung. In conclusion, the 11beta-HSD-2 enzyme of cattle is very similar to that of other species in its structure and enzymatic characteristics. PMID- 10822013 TI - Progestin regulation of 11beta-hydroxysteroid dehydrogenase expression in T-47D human breast cancer cells. AB - This study examined the enzymatic characteristics and steroid regulation of the glucocorticoid-metabolizing enzyme 11beta-hydroxysteroid dehydrogenase (11beta HSD) in the human breast cancer cell line T-47D. In cell homogenates, exogenous NAD significantly increased the conversion of corticosterone to 11 dehydrocorticosterone, while NADP was ineffective. There was no conversion of 11 dehydrocorticosterone to corticosterone either with NADH or NADPH demonstrating the lack of reductase activity. In keeping with these results, RT-PCR analysis indicated a mRNA for 11beta-HSD2 in T-47D cells, while 11beta-HSD1 mRNA levels were undetectable. In T-47D cells treated for 24 h with medroxyprogesterone acetate (MPA), 11beta-HSD catalytic activity was elevated 11-fold, while estrone (E(1)), estradiol (E(2)) and the synthetic glucocorticoid dexamethasone (DEX) were ineffective. The antiprogestin mifepristone (RU486) acted as a pure antagonist of the progestin-enhanced 11beta-HSD activity, but did not exert any agonistic effects of its own. In addition, RT-PCR analysis demonstrated that MPA was a potent inducer of 11beta-HSD2 gene expression, increasing the steady-state levels of 11beta-HSD2 mRNA. Taken together, these results demonstrate that 11beta HSD2 is the 11beta-HSD isoform expressed by T-47D cells under steady-state conditions and suggest the existence of a previously undocumented mechanism of action of progestins in breast cancer cells. PMID- 10822014 TI - Identification of twenty alternatively spliced estrogen receptor alpha mRNAs in breast cancer cell lines and tumors using splice targeted primer approach. AB - Estrogen receptor (ER) alpha splice variant transcript profiles were analyzed by RT PCR in six ER positive breast cancer cell lines, MCF-7, T47D, ZR-75, LCC1, LCC2 and LCC9, three ER negative cell lines, MDA-MB-435, MDA-MB-235 and LCC6, and three ER positive malignant breast tumors using targeted primers which specifically anneal to the splice junctions of exon 2Delta, exon 3Delta, exons 2 3Delta, exon 4Delta, exon 5Delta, exon 6Delta and exon 7Delta. The partner primers were chosen such that largest possible transcripts were amplified between exons 1 and 8. The results described here show that each splice specific primer amplified not only the single exon deleted transcript but also a number of related transcripts that have deletions in various combinations of exons. The exon 2Delta specific primer amplified five transcripts that have deletions in exon 2, exons 2 and 7, exons 2, 5, and 7, exons 2 and 4-5, and exons 2 and 4-6. The exon 3Delta specific primer amplified two transcripts that have deletions in exon 3, and exons 3 and 7. The exon 2-3Delta specific primer amplified three products that have deletions in exons 2-3, exons 2-3 and 7 and exons 2-3, 5 and 7. The exon 4Delta specific primer amplified two products that have deletions in exon 4, and exons 4 and 7. The exon 5Delta specific primer amplified three transcripts, that have deletions in exon 5, exons 5 and 2, and exons 5, and 2-3. The 6Delta specific primer amplified only one transcript that has a deletion in exon 6. The 7Delta specific primer amplified four transcripts, that have deletions in exon 7, exons 7 and 4, exons 7 and 3-4, and exons 7 and 3-5. None of the above splice specific primers amplified the wild type ER sequences. The six ER positive cell lines differed in the patterns of the variant transcripts and among the three ER negative cell lines analyzed, only MDA-MB-435 showed the presence of exon 2Delta and exon 4Delta transcripts. Analyses in the tumor samples indicated that the above transcripts are extensively modified. PMID- 10822015 TI - A novel HPLC-RIA method for the simultaneous detection of estrone, estradiol and estrone sulphate levels in breast cancer tissue. AB - Estrogen deprivation is an effective approach for treatment of hormone sensitive breast cancer. While much is known about plasma estrogen levels with respect to castration in premenopausal women and use of aromatase inhibitors in postmenopausal women, currently there is increasing interest in intra-tumour estrogen production. However, knowledge about alterations in intra-tumour estrogen levels is limited, mainly due to methodological problems with measurements of estrogen fractions in tissue samples. Here we describe a new method for simultaneous measurement of the three main estrogen fractions, estrone (E(1)), estradiol (E(2)) and estrone sulphate (E(1)S) in breast tumour tissue. Following incubation with -labelled estrogen standards, crude fractions were separated by ether extraction. The E(1)S fraction was hydrolysed with sulphatase followed by eluation on a Sephadex column. High pressure liquid chromatography (HPLC) was used to purify the individual estrogen fractions prior to RIA analysis. Estrone and E(1)S were converted into E(2), and all three estrogen fractions were finally measured by the same highly sensitive and specific radioimmunoassay using estradiol-6-(O-carboxymethyl)-oximino-2-(2--iodo histamine) as a ligand. Although several purification steps were used, the internal recovery values for tritiated estrogens were found to be 25-50% for E(1) and E(2) and 15-30% for E(1)S. The detection limit of this method was 4.3 fmol/g tissue for E(2), 19.8 fmol/g tissue for E(1) and 11.9 fmol/g E(1)S, respectively. Using tissue from locally advanced breast cancers (n = 14), we found median levels of E(1), E(2) and E(1)S to be 283.8 fmol/g tissue (range 19.8-547.5), 554.1 fmol/g (9.5-3024.2) and 209.4 fmol/g (11.9-753.4), respectively. The method described here is a promising tool to study intra-tumour estrogen fractions in breast tissue biopsies. PMID- 10822016 TI - A monoclonal antibody-based enzyme-linked immunosorbent assay of ursodeoxycholic acid 3-sulfates in human urine. AB - Sulfation of the 3-hydroxy group is assumed to be a major metabolic route of ursodeoxycholic acid (UDCA) which is used for treating various hepatobiliary diseases. We have developed a sensitive enzyme-linked immunosorbent assay (ELISA) for determining the total amount of nonamidated, glycine- and taurine-amidated ursodeoxycholic acid 3-sulfates (UDCA 3-Suls) using a newly established monoclonal antibody. In this study, 12 kinds of antibody-secreting hybridoma clones were generated by a fusion experiment between P3/NS1/1-Ag4-1 myeloma cells and the spleen cells from a BALB/c or an A/J mouse which had been immunized with a conjugate of nonamidated UDCA 3-Sul and bovine serum albumin. One of the monoclonal antibodies, Ba-10 (gamma2a, kappa), had suitable binding properties for clinical application, which was group-specific to the UDCA 3-Suls, and showed negligible cross-reactivities with various related bile acids including potentially interfering compounds, namely, the unconjugated UDCA, UDCA 7-N acetylglucosaminide, the 3-sulfates of cholic acid, chenodeoxycholic acid and deoxycholic acid. The antibody Ba-10 allowed us to develop a sensitive competitive ELISA system whose measurable range was approximately 2-200 pg per assay. A serial dilution study indicated that the ELISA enables the direct measurement of the UDCA 3-Suls in human urine before and after the administration of exogenous UDCA. The daily urinary excretion rate of UDCA 3-Suls from healthy male volunteers (n = 5) was determined to be a mean of 131 +/- 61.2 (SD) microgram as the nonamidated UDCA 3-Sul equivalent. PMID- 10822017 TI - Rapid analysis of phytoestrogens in human urine by time-resolved fluoroimmunoassay. AB - A time-resolved fluoroimmunoassay (TR-FIA), with europium labeled phytoestrogens as tracers, was developed for the quantitative determination of enterolactone, genistein and daidzein in human urine. The aim was to create a method for the screening of large populations in order to assess the possible correlations between the urinary levels and the risk of Western diseases. After the synthesis of the 5'-carboxymethoxy derivative of enterolactone and 4'-O-carboxymethyl derivatives of daidzein and genistein, the respective compound was coupled to bovine serum albumin and then used as an antigen in the immunization of rabbits. The same derivatives of the phytoestrogen were used in preparing the europium tracers. After the enzymatic hydrolysis, the TR-FIA was carried out using the Victor 1420 multilabel counter. The method has sufficient sensitivity to measure the phytoestrogens at concentrations even below 5 nmol/l. The intra- and inter assay coefficients of variation, at three different concentrations, varied from 1.9 to 5.3 and from 2.4 to 9.7, respectively. We measured urinary enterolactone, genistein and daidzein in 215 samples from Finnish healthy women and found that more than 50% of the values ranged between 1 and 7, <0.1 and 0.6 and below 0.6 micromol/24 h, respectively. The TR-FIA method including only a hydrolysis step gave higher values than those measured by gas chromatography-mass spectrometry (GC-MS). However, the assay results by the present method showed strong correlation with those obtained by GC-MS. It is concluded that the TR-FIA is suitable for population screening of urinary phytoestrogens. PMID- 10822018 TI - Effect of ispaghula husk on the faecal output of bile acids in healthy volunteers. AB - Faecal bile acids are associated with both colorectal cancer and serum cholesterol levels. We investigate whether dosing with ispaghula husk affects the faecal bile acid weights and concentrations in healthy adults. Sixteen healthy volunteers consumed 7.0 g/day ispaghula husk, containing 5.88 g/day Englyst determinable dietary fibre, for the middle 8 weeks of a 12-week period. Stool samples were collected, analysed for faecal bile acids and their form and dry weight determined. Correlations between the faecal bile acids, the stool parameters and the dietary intake were tested. Ispaghula husk treatment significantly lowers faecal lithocholic and isolithocholic acids and the weighted ratio of lithocholic acids to deoxycholic acid. These effects revert towards their initial states at the end of the treatment period. These changes in the faecal bile acid profiles indicate a reduction in the hydrophobicity of the bile acids in the enterohepatic circulation. PMID- 10822019 TI - Ability of xeno- and phytoestrogens to modulate expression of estrogen-sensitive genes in rat uterus: estrogenicity profiles and uterotropic activity. AB - The function of the uterus is regulated by female sex steroids and it is, therefore, used as the classical target organ to detect estrogenic action. Uterine response to estrogens involves the activation of a large pattern of estrogen-sensitive genes. This fact offers the opportunity to analyze the estrogenic activity of xeno- and phytoestrogens, and the mechanisms of their molecular action by a correlation of the uterotropic activity and their ability to modulate the expression of estrogen-sensitive genes. We have analyzed the expression of androgen receptor (AR), progesterone receptor (PR), estrogen receptor (ER), clusterin (CLU), complement C3 (C3), and GAPDH mRNA in the rat uterus following oral administration of ethinylestradiol (EE), bisphenol A (BPA), o,p'-DDT (DDT), p-tert-octylphenol (OCT) and daidzein (DAI). A significant stimulation of the uterine wet weight could be observed after administration of all the substances. The activity of all analyzed compounds to stimulate uterine weight was low in comparison to EE. DDT has the highest activity to stimulate uterine weight whereas BPA and DAI turned out to be less potent. The analysis of gene expression revealed a very specific profile of molecular action in response to the different compounds which cannot be detected by judging the uterotropic response alone. A dose dependent analysis revealed that C3 mRNA is already modulated at doses where no uterotropic response was detectable. Although DAI and BPA were very weak stimulators of uterine growth, these substances were able to alter the expression of AR, ER and C3 very strongly. Based on these investigations the analyzed compounds can be subdivided into distinct classes: First, compounds which exhibit a similar gene expression fingerprint as EE (e.g. OCT); second, compounds exhibiting a significant uterotropic activity, but inducing a pattern of gene expression different from EE (e.g. DDT); and third, compounds like BPA and especially DAI which exhibit a very low uterotropic activity, but nevertheless modulate the expression of estrogen-sensitive genes. These findings strongly suggest that the fingerprint of uterine gene expression is a very sensitive tool to investigate estrogenicity of natural and synthetic compounds and offers the possibility to get information in regard to the molecular mechanisms involved in the action of the respective compounds. PMID- 10822020 TI - Aldosterone alters the phospholipid composition of rat colonocytes. AB - Previous studies have shown that aldosterone treatment of amphibian epithelial cells results not only in stimulation of Na(+) absorption but also in changes in phospholipid composition which are necessary for the mineralocorticoid action of aldosterone. The present study was designed to investigate the effect of aldosterone on phospholipids of mammalian epithelia. Phospholipid and fatty acid composition was examined in colonic epithelium (mineralocorticoid target tissue) and thymus (non-mineralocorticoid but glucocorticoid target tissue) of rats which had received aldosterone or vehicle by a miniosmotic pump for 7 days. Aldosterone increased the mass of colonic phospholipids relative to cellular proteins with concomitant changes in the percentage distribution of fatty acids, whereas the relative distribution of membrane phospholipds was not changed. Phosphatidylcholine increased the content of polyunsaturated and decreased that of monounsaturated fatty acids, which predominantly reflected the accretion of arachidonic and a decrease in oleic and palmitoleic acids. Within the phosphatidylethanolamine subclass, pretreatment of rats with aldosterone decreased the content of monounsaturated fatty acids (predominantly oleic and palmitoleic acid) and of n-3 fatty acids, and increased the content of saturated fatty acids (palmitic acid). The saturated-to-nonsaturated fatty acid ratio also significantly increased after aldosterone treatment. No changes in thymic phospholipids were seen. The results are consistent with the contention that aldosterone specifically modulates phospholipid concentration and metabolism in mineralocorticoid target tissue. The changes in phospholipid content and its fatty acid composition during the fully developed effect of aldosterone may reflect a physiologically important phenomenon with long-term consequences for membrane structure and function. PMID- 10822021 TI - Absence of correlation between in situ expression of cytochrome P450 17alpha hydroxylase/lyase and 3beta-hydroxysteroid dehydrogenase/(Delta5-4) isomerase messenger ribonucleic acids and steroidogenesis during pubertal development in the rat testis. AB - Changes in expression of Leydig cell 3beta-hydroxysteroid dehydrogenase (3betaHSD) and 17alpha-hydroxylase/C17-20 lyase (P450(17alpha)) messenger RNA (mRNA) during pubertal development have not been well characterized in the rat. In the present study, expression of 3betaHSD and P450(17alpha) were determined in frozen sections of testes of immature (days 21 and 28), pubertal (days 45 and 60) and adult (day 90) rats by in situ hybridization using digoxigenin-labeled riboprobes and quantified densitometrically. Measures of steroidogenesis in this study, 3betaHSD and P450(17alpha) enzyme activities per testis and plasma testosterone concentration, increased during pubertal development, peaking at 45 60 days of age. Expression of 3betaHSD protein, a marker for Leydig cell function, was abundantly immunolocalized to the interstitial compartment of the testis. Quantified densitometrically, the amount of 3betaHSD protein did not vary significantly during pubertal development. Transcripts of 3betaHSD and P450(17alpha) were expressed abundantly by clusters of immature Leydig cells in immature animals. However, in contrast to measures of steroidogenesis during pubertal development, mRNA of 3betaHSD and P450(17alpha) decreased to undetectable levels at the age of 45 and 60 days, respectively. The decline in mRNA of 3betaHSD and P450(17alpha) was confirmed by Northern analysis. Expression of 3betaHSD and P450(17alpha) transcripts rebounded in the adult at 90 days and were comparable to levels of expression observed in immature animals. These results show that during pubertal development the steady-state accumulation of mRNA of 3betaHSD and P450(17alpha) are not correlated with accumulation of 3betaHSD protein, enzyme activities of 3betaHSD and P450(17alpha), or testosterone secretion. Possible explanations of the depletion of transcripts during pubertal development include: specific inhibition of transcription, increased mRNA instability, or high translational activity. PMID- 10822022 TI - Response of estrogen receptor containing tumour cells to pure antiestrogens and the calmodulin inhibitor, calmidzolium chloride. AB - Cell survival is dependent on both external and internally generated signalling processes and current strategies for medical intervention in neoplastic disease are directed towards signal transduction blockade. Redundancy in signalling pathways may mean, however, that a combination of agents is required for the maximal therapeutic benefit. We have explored this idea with regard to the antiestrogen sensitivity of estrogen dependent tumours. Using estrogen receptor (ER) containing tumour cell lines, we have determined whether antiestrogens increase the cytotoxicity of the potent calmodulin inhibitior, calmidzolium chloride (CCl). For the pituitary tumour cell line GH(3), CCl induces a form of apoptotic cell death and co-treatment with the pure antiestrogen, ZM 182780, enhances sensitivity to the calmodulin inhibitor, by at least two fold. In contrast to the pure steroidal antiestrogens, the triphenylethylenes, tamoxifen and 4-hydroxytamoxifen give no enhancing effect on CCl induced cell death. Although CCl induces apoptosis of several ER containing breast cancer cell lines, unlike the pituitary tumour cells, ZM 182780 is unable to increase their sensitivity to calmodulin inhibition. Further studies strongly suggest that cell death in response to calmodulin inhibition is the result of metabolic disruption and that for GH(3) cells, this is enhanced by antiestrogen treatment. PMID- 10822023 TI - Excretion balance and metabolism of the progestagen Org 30659 in healthy postmenopausal women. AB - Metabolism of Org 30659 ((17alpha)-17-hydroxy-11-methylene-19-norpregna-4, 15 dien-20-yn-3-one), a new potent progestagen currently under clinical development by NV Organon for use in oral contraception and hormone replacement therapy, was studied in vivo after oral administration to healthy postmenopausal women. After oral administration of [14C]-Org 30659 to postmenopausal women, the compound was extensively metabolized. The dosed radioactivity was predominantly excreted via urine. Org 30659 was to a large extent metabolized at the C3- and the C17 positions. Phase II metabolism, and in particular conjugation with glucuronic acid at the 17beta-hydroxy group, is the major metabolic route for Org 30659 in vivo. Not only phase II metabolism was observed for Org 30659 after oral administration to postmenopausal volunteers, but also metabolism in the A-ring occurred, especially reduction of the 3-keto-Delta(4) moiety to give 3alpha hydroxy, 5alpha(beta)-dihydro and 3beta-hydroxy, 5alpha-dihydro derivatives. Oxidative metabolism (6beta-hydroxylation) observed in human liver preparations in vitro, was not observed to a significant extent in vivo. So, in vitro human metabolism is different from the in vivo metabolism, indicating that the in vitro in vivo extrapolation is far from straightforward, at least when only liver preparations are used. The proper choice of the in vitro system (e.g., microsomes, hepatocytes, slices or individually expressed enzymes) and the substrate concentration can be very important determinative factors for the predictability of the in vitro system for the in vivo situation. Species comparison of the metabolic routes of Org 30659 after oral administration indicated that the monkey seems to be a better representative species than the rat for the metabolism of Org 30659 in humans. PMID- 10822024 TI - Multiple isoforms of porcine aromatase are encoded by three distinct genes. AB - Cytochrome P450 aromatase, a product of the CYP 19 gene and the terminal enzyme in the estrogen biosynthetic pathway, is synthesized by the ovary, endometrium, placenta, and peri-implantation embryos in the pig and other mammals, albeit to varying levels, implying its functional role(s) in pregnancy events. The aromatase produced by the pig tissues exists as three distinct isoforms (type I - ovary, type II - placenta, and type III - embryo), with presumed differences in substrate specificities, expression levels, activity, and mode of regulation. In order to delineate the molecular mechanisms whereby estrogen synthesis is regulated in these diverse tissues, the present study examined if these aromatase isoforms represent products of multiple genes or of a single gene via complex splicing mechanisms. Porcine genomic DNA from a single animal was used as a template in the polymerase chain reaction (PCR) to amplify isoform-specific sequences corresponding to exons 4 and 7, respectively. Nucleotide sequence analysis of the generated fragments revealed the presence of only clones corresponding to the three known aromatase types. Screening a porcine Bacterial Artificial Chromosome (BAC) library for aromatase gene by PCR yielded a single clone approximately 80 kb in length. Southern blot analysis, using probes specific for exons 1A-1B, 2-3, 4-9, and 10 sequences indicated that the BAC genomic clone contains the entirety of the coding exons as well as the proximal promoter region. Sequence analysis of the fragment generated with exon 4 primers determined that this BAC clone contains only the type II gene. The presence and relative orientation of the untranslated 5'- exons 1A and 1B, previously demonstrated for the type III isoform were evaluated in the BAC clone and genomic DNA by PCR. The 265 bp fragment generated from both PCR reactions was confirmed by sequence analysis to contain exons 1A and 1B that are located contiguous to each other and separated by only three bp. A diagnostic procedure for typing aromatase isoforms was developed, based on the presence of specific restriction sites within isoform-specific exons. The use of this protocol confirmed the existence of only three aromatase isoforms in the porcine genome and indicated changes in aromatase types expressed by the uterine endometrium as a function of pregnancy stage. The presence of distinct genes encoding each of the aromatase isoform predicts important differences in the mechanisms underlying the molecular evolution and regulation of porcine aromatase, unique from those of other mammals, and suggests a critical role for P450 aromatase steroidal products in uterine functions related to pregnancy events. PMID- 10822025 TI - In vivo estrogen bioactivities and in vitro estrogen receptor binding and transcriptional activities of anticoagulant synthetic 17beta-aminoestrogens. AB - Estrogenic activities of the two 17beta-aminoestrogen (AE) derivatives, prolame and butolame, were studied upon coagulation, serum luteinizing hormone (LH) and uterine weight, including endometrial morphology in castrated female rats. We have also investigated the ability of these two compounds, as well as another AE pentolame, to activate transcription through the estrogen receptor alpha (ERalpha) and the estrogen receptor beta (ERbeta). Administration of prolame and butolame to castrated animals increased significantly (P < 0.01) the mean clotting time when compared with that obtained in the group of control animals. Butolame was a more potent anticoagulant than prolame (P < 0.01), as judged by their corresponding IC(50) (5.4 +/- 0.65 and 66.6 +/- 2.57 micro;g/animal, respectively). In contrast, estradiol significantly shortened blood clotting times (P < 0.005). Both prolame and butolame caused a significant inhibition of serum LH levels (EC(50) 8.10 +/- 0.79 and 17 +/- 64 microg/animal, respectively), and restored castration-induced reduction in uterine weight of ovariectomized rats (EC(50) 4.14 +/- 1.57 and 17.0 +/- 1.78 microg/animal, respectively). In terms of the effects of prolame, butolame and pentolame in transient transfection assays, all the three AE activated ER dependent reporter gene expression, however, only at high concentrations. Prolame had the highest activity followed by butolame and pentolame. Induction of transcription by these compounds was preferentially mediated through the ERalpha, especially in the case of pentolame where little, if any, activation occurred through the ERbeta. None of the compounds showed antagonistic activities through either ER subtype. The overall data suggest that modifications in the structure and length of the amino-alcohol side-chain at C-17 might have an impact on the affinity and estrogenic intrinsic properties of AE at the level of diverse target tissues. PMID- 10822026 TI - Effect of natural "micronized" progesterone on the chorionic gonadotropin concentrations in cyst fluids of women with gross cystic breast disease. AB - Gross cystic breast disease (GCBD) is common in women, especially in the age range between 35 and the menopausal years. The present study examined the possible role of progesterone (Pg) in the chorionic gonadotropin (hCG) concentration in GCBD. The breast cyst fluids (BCFs) were drawn by fine needle aspiration between the sixth and the eighth day of the menstrual cycle and twenty days later. On the day of the first aspiration the patient began to take 100 mg of natural micronized Pg orally until the second aspiration. At both times blood samples were also taken. Determinations were done of both BCFs and blood sample using two fully automated chemiluminiscent enzyme immunometric assays. Pg has been demonstrated to induce a significant increment in hCG + free ss-hCG (median, range): 0.27 ng/ml, 0.12-6.24 vs. 1.92 ng/ml, 0.12-423.5; free ss-hCG: 0.11 ng/ml, 0.02-2.40 vs. 0.91 ng/ml, 0.02-58.40 in the BCFs, with no change in the circulating concentrations of the hormone. None of the sera studied presented levels of hCG + free ss-hCG or free ss-hCG above 0.5 ng/ml or 0.1 ng/ml, respectively. The occurrence of hCG or a derivative polypeptide in BCFs, when they are present in high concentrations suggests that this glycoprotein could be synthesized in situ and possibly involved in the pathogenesis of GCBD by the degree of differentiation of breast epithelial cells induced by the hormone. PMID- 10822027 TI - Role of rat adrenal antioxidant defense systems in the aldosterone turn-off phenomenon. AB - The mechanism(s) of the "aldosterone turn-off phenomenon", hypoaldosteronemia following chronic ACTH administration, remains unclear. Our previous observation that antioxidants prevented turn-off prompted us to evaluate the chronic effect of ACTH on the enzymatic antioxidant system as well as P450aldo activity and expression of CYP11B2 in adrenal zona glomerulosa. Male Wistar rats were administered ACTH-Z for 5 days with or without antioxidants, vitamin E or DMSO. Adrenal capsules were prepared for P450aldo activity measurement and mRNA content determination by competitive RT-PCR, and immunoreactivity of Mn-SOD in whole adrenals was evaluated. ACTH decreased the P450aldo activity and mRNA level of CYP11B2 in adrenal capsules, while co-administration of vitamin E or DMSO partially blocked this inhibition. ACTH increased Mn-SOD mRNA and immunoreactivity but decreased GPx mRNA. These results suggest that prolonged ACTH treatment increases oxidative stress in the zona glomerulosa and an imbalance in the ratio of Mn-SOD to GPx, possibly via corticosterone overproduction in the zona fasciculata, resulting in the downregulation of CYP11B2. Vitamin E and DMSO might thus protect CYP11B2 expression through their antioxidant actions. PMID- 10822028 TI - Cyclic-AMP-dependent protein kinase (PKA) in testicular cells. Cell specific expression, differential regulation and targeting of subunits of PKA AB - LH and FSH regulate via cyclic adenosine 3'5' cyclic monophosphate (cAMP) and cAMP-dependent protein kinase (PKA), steroid biosynthesis is Leydig and Sertoli cells, respectively. Cyclic AMP also regulates a number of different cellular processes such as cell growth and differentiation, ion channel conductivity, synaptic release of neurotransmitters, and gene transcription. The principle intracellular target for cAMP in mammalian cells is the PKA. The fact that this broad specificity protein kinase mediates a number of discrete physiological responses following cAMP engagement, has raised the question of how specificity is maintained in the cAMP/PKA system. Here we describe features of this signaling pathway that may contribute to explain how differential effects of cAMP may occur. PMID- 10822035 TI - Management of atrial fibrillation: therapeutic options and clinical decisions. AB - Atrial fibrillation (AF) is the most common, sustained tachyarrhythmia seen in clinical practice. Although it is not immediately life threatening, AF can cause troublesome symptoms and poses a risk of stroke. The patient's clinical status is often complicated by the presence of other cardiovascular or concomitant diseases. As a result, management of the patient with AF involves many questions and choices, all of which must be individualized. There are 3 general strategies for the management of patients with AF, including (1) restoration and maintenance of sinus rhythm, (2) control of ventricular rate, and (3) prevention of stroke. More than 1 strategy may be appropriate in some patients. Furthermore, either pharmacologic or nonpharmacologic options can be chosen in certain situations. Although some data from randomized clinical trials are available to aid in clinical decision-making, only the benefits of anticoagulation are supported by substantial evidence. This article explores practical approaches to several management issues and scenarios for which there are limited relevant clinical data. These include: (1) patient selection for ventricular rate control and assessment of treatment, (2) choice of antiarrhythmic drug for maintenance of sinus rhythm, (3) inpatient versus outpatient initiation of therapy, (4) definition of antiarrhythmic drug success, (5) methods of transthoracic direct cardioversion, and (6) prediction and prevention of AF after cardiac surgery. PMID- 10822036 TI - Drug choices in the treatment of atrial fibrillation. AB - When considering therapy for atrial fibrillation (AF), the dominant issues are rate control, anticoagulation, rhythm control, and treatment of any underlying disorder. Drug choices for rate control include beta-blockers, verapamil and diltiazem, and digitalis as first-line agents, with consideration of other sympatholytics, amiodarone, or nonpharmacologic approaches in resistant cases. Anticoagulation may be accomplished with aspirin or warfarin, with the latter preferred in all older or high-risk patients. Antiarrhythmic drug therapy may be used (1) to produce cardioversion (most effective with ibutilide or class IC agents in recent onset AF); (2) to facilitate electrical conversion (class III agents); (3) to prevent early reversion after cardioversion; (4) to maintain sinus rhythm during chronic therapy; and/or (5) to facilitate conversion of fibrillation to flutter, which may then be amenable to termination or prevention with antitachypacing or ablative techniques. Antiarrhythmic drug selection for AF is guided by efficacy considerations (most drugs are similar), by convenience, cost, and discontinuation considerations; and, most importantly, by safety considerations. When possible, agents with serious organ toxicity potential and proarrhythmic risk should be avoided as first-line choices. In structurally normal hearts, class IC antiarrhythmic drugs are least proarrhythmic and least organ toxic (when considered together). In normal hearts, sotalol, dofetilide, and potentially azimilide also appear to have attractive profiles. Amiodarone has low proarrhythmic risk but can produce bradyarrhythmias and toxicity. In hypertrophied hearts, the risk of torsade de pointes with class III/IA agents is enhanced, whereas in ischemia or conditions with impaired cell contact, whether functionally (as by ischemia) or anatomically (as by fibrosis, infiltration, etc), proarrhythmic risk with class I antiarrhythmic drugs (sustained ventricular fibrillation/flutter) is greatly increased. The class I drugs should be avoided in these circumstances. Additional issues to consider are where to initiate therapy (in- or outpatient), what follow-up protocols to use, and whether to limit therapy to proprietary drugs or to allow generic formulation substitution. Each of these considerations is detailed in this article. PMID- 10822037 TI - Management and prevention of atrial fibrillation after cardiovascular surgery. AB - Atrial arrhythmias occur frequently after cardiac surgery. This article discusses the incidence of postoperative atrial arrhythmia as well as its prognosis, potential mechanisms of pathogenesis, and management. Prophylactic therapy for postoperative atrial arrhythmia is recommended because of the frequency of occurrence and the ease with which therapies can often be implemented. Treatments with pharmacologic and nonpharmacologic modalities are described. Management strategies for atrial arrhythmias that occur postoperatively, including pharmacologic and nonpharmacologic measures as well as anticoagulation recommendations, are discussed. PMID- 10822038 TI - Atrial fibrillation: nonpharmacologic approaches. AB - Various nonpharmacologic interventions are available for patients with atrial fibrillation (AF) who are refractory to standard drug therapy. Atrioventricular junctional ablation and permanent pacing is a very effective therapy for patients with AF and a poorly controlled ventricular response. The surgical MAZE procedure has been performed on small numbers of patients but is remarkably successful in restoring and maintaining sinus rhythm. The role of permanent pacing as treatment for paroxysmal AF is undergoing evaluation and dual-site atrial pacing appears particularly promising in reducing the number of episodes of paroxysmal AF. Certainly the most exciting frontier in the treatment of AF is radiofrequency catheter ablation procedures. Our understanding of the mechanisms of paroxysmal AF and chronic AF has expanded enormously in the past 5 years. Radiofrequency lesions in pulmonary veins using standard technology will cure many cases of paroxysmal AF. However, catheter systems under development offer a great promise of treating most paroxysmal and chronic AF within the next few years. These developments will revolutionize our approach to this ever more prevalent clinical problem. PMID- 10822039 TI - Cost-effective management of acute atrial fibrillation: role of rate control, spontaneous conversion, medical and direct current cardioversion, transesophageal echocardiography, and antiembolic therapy. AB - Management strategies for the acute treatment of atrial fibrillation (AF) include: (1) the use of intravenous drugs for rate control, (2) drug termination, or (3) direct current (DC) cardioversion. Delays in cardioversion can promote atrial remodeling and add thromboembolic risk. Rate control awaiting spontaneous or pharmacologic conversion may be a cost-effective strategy in patients presenting with recent onset of symptoms. Early DC cardioversion can be cost effective and minimize antiembolic therapy issues in the acute setting. In patients presenting with AF of unknown or >48 hours' duration, rate control and therapeutic warfarin for 3-4 weeks followed by medical or DC cardioversion is standard practice. However, delays in conversion promote atrial remodeling that makes restoration of sinus rhythm more difficult and increases the likelihood of postcardioversion AF recurrence. Transesophageal echocardiography can identify patients at low risk for a cardioversion-related embolic event and allows cardioversion to be performed earlier, thereby minimizing atrial remodeling. PMID- 10822040 TI - Formulation substitution and other pharmacokinetic variability: underappreciated variables affecting antiarrhythmic efficacy and safety in clinical practice. AB - The process of treating a patient with an antiarrhythmic drug only begins when a physician chooses the drug to be employed. In the given patient, not only must the drug be chosen, but so must the dose, the formulation, and the method of follow-up. Choosing the proper dose requires an understanding of clinical trial efficacy and safety data for the agent chosen in a population of patients resembling the individual to be treated. It also requires a detailed understanding of pharmacologic principles of drug kinetics (e.g., absorption, distribution, metabolism, and excretion) that might affect the dose needed for the specific patient. The physician must be familiar with subsequent changes in clinical circumstances that might indicate a need for a change in dose or drug. Many circumstances determining drug pharmacokinetics are not under the immediate control of physicians, such as genetic patterns, organ function, and disease circumstances. One, however, is-or should always be-the selection of the drug formulation used. Although generic versions of innovator drugs exist for many agents and often are clinically acceptable, most physicians are unaware of the meager degree of testing that is necessary for the release of a generic drug, and the wide range of attained serum levels that are called bioequivalent by the US Food and Drug Administration (FDA) when one formulation is compared with another. In patients with cardiac arrhythmias, arrhythmia recurrence, proarrhythmia, and death have been reported in association with antiarrhythmic drug formulation substitution. Despite their reported bioequivalence, the generic agents involved were clearly not therapeutically equivalent. Accordingly, this article was written to educate physicians further about the above-noted important pharmacokinetic variables that can affect a patient's outcome when an antiarrhythmic drug is employed, and to provide information on the generic drug approval process and guidelines for the use of formulation substitution. PMID- 10822041 TI - Regulation of division of labour between cognitive systems controlling action. AB - The brain has evolved a division of labour amongst component systems which link different sorts of processing with precise actions. Debate is over centralized versus decentralized control at different processing levels, from cognitive systems to motor-control systems. With simultaneous activation of alternative expert systems which link (a) picture-processing with drawing and (b) reading with writing, decentralized modelling predicted both the averaging of action production times and additive effects of neural noise. Such modelling has the advantage of being able to measure the cost of regulation both within and between systems, in a common metric of performance variability. That commonality strengthens the trend to regard the brain as a distributed super-system with a great deal of regulation done towards the output end. PMID- 10822042 TI - Cross-cultural music cognition: cognitive methodology applied to North Sami yoiks. AB - This article is a study of melodic expectancy in North Sami yoiks, a style of music quite distinct from Western tonal music. Three different approaches were taken. The first approach was a statistical style analysis of tones in a representative corpus of 18 yoiks. The analysis determined the relative frequencies of tone onsets and two- and three-tone transitions. It also identified style characteristics, such as pentatonic orientation, the presence of two reference pitches, the frequency of large consonant intervals, and a relatively large set of possible melodic continuations. The second approach was a behavioral experiment in which listeners made judgments about melodic continuations. Three groups of listeners participated. One group was from the Sami culture, the second group consisted of Finnish music students who had learned some yoiks, and the third group consisted of Western musicians unfamiliar with yoiks. Expertise was associated with stronger veridical expectations (for the correct next tone) than schematic expectations (based on general style characteristics). Familiarity with the particular yoiks was found to compensate for lack of experience with the musical culture. The third approach simulated melodic expectancy with neural network models of the self-organizing map (SOM) type (Kohonen, T. (1997). Self-organizing maps (2nd ed.). Berlin: Springer). One model was trained on the excerpts of yoiks used in the behavioral experiment including the correct continuation tone, while another was trained with a set of Finnish folk songs and Lutheran hymns. The convergence of the three approaches showed that both listeners and the SOM model are influenced by the statistical distributions of tones and tone sequences. The listeners and SOM models also provided evidence supporting a core set of psychological principles underlying melody formation whose relative weights appear to differ across musical styles. PMID- 10822043 TI - A componential view of theory of mind: evidence from Williams syndrome. AB - In this paper we argue that there are two distinct components of a theory of mind: a social-cognitive and a social-perceptual component. Evidence for this proposal is presented from various sources, including studies of children with Williams syndrome, a rare genetic neurodevelopmental disorder. Earlier work has demonstrated that people with Williams syndrome appear to be spared in the social perceptual component of a theory of mind. In this paper we present evidence that they are not spared in the social-cognitive component of theory of mind. Three experiments with young children with Williams syndrome were conducted. In each experiment the children with Williams syndrome were compared to age-, IQ-, and language-matched children with Prader-Willi syndrome, and children with non specific mental retardation. The experiments used different measures of theory of mind ability, including false belief (Experiment 1), explanation of action (Experiment 2), and recognition of emotional expressions (Experiment 3). In none of these experiments did the children with Williams syndrome evidence superior performance compared to the control groups. The results from this and other studies on Williams syndrome support the view that the social-cognitive and social-perceptual components of a theory of mind are dissociable. In Williams syndrome only the latter components, which are linked to distinct neurobiological substrates, are spared. PMID- 10822044 TI - Reading Chinese characters for meaning: the role of phonological information. AB - Two experiments with the Stroop paradigm were conducted to investigate the role of phonological codes in access to the meaning of Chinese characters. Subjects named the ink color of viewed characters or color patches. Key items were color characters, their homophones with the same tone, homophones with different tones, and semantic associates. Apart from finding the usual Stroop interference effect, homophones produced significant interference in the incongruent condition, provided that they had the same tone as the color characters. The interference effect from homophones, however, was significantly smaller than that from color characters. Semantic associates generated an interference effect in the incongruent condition, an effect of the same magnitude as the effect from the same-tone homophones. Finally, in the congruent conditions, all the key items yielded facilitations compared to neutral controls, though the facilitation from color characters was larger than the facilitations from other types of characters. These findings suggest that phonological codes in Chinese are activated obligatorily and provide early sources of constraint in access to meaning. PMID- 10822045 TI - The rapid use of gender information: evidence of the time course of pronoun resolution from eyetracking. AB - Eye movements of listeners were monitored to investigate how gender information and accessibility influence the initial processes of pronoun interpretation. Previous studies on this issue have produced mixed results, and several studies have concluded that gender cues are not automatically used during the early processes of pronoun interpretation (e.g. Garnham, A., Oakhill, J. & Cruttenden, H. (1992). The role of implicit causality and gender cue in the interpretation of pronouns. Language and Cognitive Processes, 73 (4), 231-255; Greene, S. B., McKoon, G. & Ratcliff, R. (1992). Pronoun resolution and discourse models. Journal of Experimental Psychology: Learning, Memory, and Cognition, 182, 266 283). In the two experiments presented here, participants viewed a picture with two familiar cartoon characters of either same or different gender. They listened to a text describing the picture, in which a pronoun referred to either the first, more accessible, character, or the second. (For example, Donald is bringing some mail to ?Mickey/Minnie? while a violent storm is beginning. He's carrying an umbrellaellipsis.) The results of both experiments show rapid use of both gender and accessibility at approximately 200 ms after the pronoun offset. PMID- 10822046 TI - Antagonist activity of meta-chlorophenylpiperazine and partial agonist activity of 8-OH-DPAT at the 5-HT(7) receptor. AB - This study compared the use of adapter G-proteins to link G(s) coupled G-protein receptors to a Ca(2+) signal, enabling high throughput functional studies using a fluorescent imaging plate reader (FLIPR, Molecular Devices). The pharmacological profile of the human 5-hydroxytryptamine (5-HT(7)) receptor was studied using the adapter G-proteins G(alpha16) and G(qs5) and compared to previously published adenylyl cyclase and receptor binding data. Human embryonic kidney (HEK) 293 cells stably expressing the human 5-HT(7(a)) receptor were transiently transfected with the adapter G-proteins. Changes in intracellular Ca(2+) were monitored using the fluorescent Ca(2+)-indicator Fluo-4.5-Carboxamidotryptamine (5-CT) induced an increase in fluorescence in transfected cells only, which was attenuated by N-ethylmalaeimide and abolished by thapsigargin, consistent with a G-protein mediated mobilisation of intracellular Ca(2+). The pharmacological profile of agonists at the 5-HT(7) receptor was similar using either adapter G protein. Agonist potency estimates were similar to that reported in binding studies but were greater than that seen in adenylyl cyclase studies. 8-Hydroxy-N, N-dipropylaminotetralin (8-OH-DPAT) and tryptamine acted as partial agonists using the adapter G-proteins, but were full agonists in recombinant systems using adenylyl cyclase. meta-Chlorophenylpiperazine (mCPP) and trifluoro-methylphenyl piperazine (TFMPP) were antagonists on intracellular Ca(2+). Antagonist pharmacological profiles were similar between adapter G-proteins, receptor binding, and adenylyl cyclase studies. These results show that adapter G-proteins can be used to study G(s)-linked receptors using the high throughput FLIPR system measuring changes in intracellular Ca(2+) and provide novel information on mCPP and 8-OH-DPAT. PMID- 10822047 TI - Cloning of rabbit alpha(1b)-adrenoceptor and pharmacological comparison of alpha(1a)-, alpha(1b)- and alpha(1d)-adrenoceptors in rabbit. AB - We have isolated a cDNA clone of the rabbit alpha(1b)-adrenoceptor which has an open reading frame of 1557 nucleotides encoding a protein of 518 amino acids. The sequence shows higher identity to those of hamster, human, and rat alpha(1b) adrenoceptors than to those of rabbit alpha(1a)- and alpha(1d)-adrenoceptors. The pharmacological binding properties of this clone expressed in Cos-7 cells showed a characteristic profile as alpha(1b)-adrenoceptor; high affinity for prazosin (pK(i)=10.3), relatively high affinity for tamsulosin (9.5) and low affinity for (-)-(R)-1-(3-hydroxypropyl)-5-[2-[[2-[2-(2,2, 2 trifluoroethoxy)phenoxy]ethyl]amino]propyl]indoline-7-carboxamid e (KMD3213) (8.5), 2-(2,6-dimethoxy-phenoxyethyl)-aminomethyl-1, 4-benzodioxane hydrochloride (WB4101) (8.7), and 8-[2-[4-(2-methoxy-phenyl)-L-piperazinyl]-8 azaspiro[4,5]decane-7, 9-dione dihydrochloride (BMY7378) (7.3). We have compared the levels of mRNA expression of three alpha(1)-adrenoceptor subtypes in rabbit tissues using the competitive reverse transcription/polymerase chain reaction (RT/PCR) assay. In most rabbit tissues except heart, alpha(1a)-adrenoceptor mRNA was expressed 10 folds more than the other two subtypes. However, binding experiments with [3H]prazosin and [3H]KMD3213 in rabbit tissues revealed a poor relationship between binding density and mRNA level. Especially, alpha(1b) binding sites were exclusively predominant in spleen, whereas the alpha(1b) subtype was minor at the mRNA level. These results indicate a high identity of structural and pharmacological profiles of three distinct alpha(1)-adrenoceptor subtypes between rabbit and other species, but there are species differences in their distribution. PMID- 10822048 TI - Enhanced endothelin ET(B) receptor down-regulation in human tumor cells. AB - The characteristics of specific binding of human [125I]Tyr(13)-endothelin-(1-21), [125I]-Tyr(13)-Suc-[Glu(9),Ala(11, 15)]-endothelin-(8-21), ([125I]IRL-1620) and endothelin ET(A) receptor antagonist [125I]Tyr(3)-(N-[(hexahydro-1H-azepin-1 yl)carbonyl]-L-Leu]-1Me )-D-Trp ([125I]PD151242) (number of sites and their affinity) and proliferation responses to exogenous endothelin receptor agonists (endothelin-1 and the endothelin ET(B) receptor-selective, truncated N-acetyl [Ala(11,15)]-endothelin-(6-21) analogue BQ3020) were determined in cultured human fibroblasts and in tumorigenic HeLa cells. The cells were pre-incubated with equimolar concentrations of human endothelin-1 or its truncated analogue BQ3020. After pre-incubation (2 h), both peptides induced down-regulation of surface membrane endothelin-1 receptors. This process was specific for endothelin ET(B) receptors and was much more intensive in tumorigenic cells. BQ3020, acting mostly through its C-terminus, induced nearly maximal endothelin ET(B) receptor down regulation in HeLa cells. Staurosporine, a wide spectrum protein kinase inhibitor, significantly reduced, and N-[N-[N-[2, 6-dimethyl 1piperidinyl)carbonyl]-4-Me-L-Leu]-1-(methoxycarbonyl)- D-t ryptophanyl]-D norleucine (BQ788), an endothelin ET(B) receptor antagonist, attenuated the down regulation of endothelin receptors induced by endothelin receptor agonists. The down-regulation of endothelin ET(B) receptors was prevented by pre-incubation of the cells with the lysosomal enzyme blocker chloroquine. The endothelin-1-induced cell proliferation was attenuated by pre-incubation of the cells with the non selective endothelin receptor antagonist Ac-D-10,11-dihydro-5H-dibenzo[a,d] cycloheptene-glycine-3,3-D-diphenyl-Ala-Leu-Asp-Ile-Ile-Trp (PD142893) and it was only partially reduced by the endothelin ET(A) receptor-selective endothelin antagonist PD151242. PMID- 10822050 TI - Tachykinin NK(2) receptors facilitate acetylcholine release from guinea-pig isolated trachea. AB - The release of newly synthesised [3H]acetylcholine was evoked by electrical field stimulation (5 Hz, 600 pulses) of epithelium-deprived guinea-pig trachea strips after sensory neuropeptides depletion with 3 microM capsaicin. The selective tachykinin NK(2) receptor agonist [betaAla(8)]neurokinin A-(4-10) increased in a concentration-dependent manner the electrically-induced release of [3H]acetylcholine. The facilitatory effect was antagonised by the selective non peptide tachykinin NK(2) receptor antagonist, SR 48968 (apparent pK(B) 8.9). The tachykinin NK(1) and NK(3) receptor agonists substance P methyl ester and senktide (both 10 and 100 nM), respectively, did not affect the evoked release of [3H]acetylcholine. It is concluded that the cholinergic nerves of guinea-pig trachea are endowed with prejunctional facilitatory tachykinin receptors of the NK(2) subtype. PMID- 10822049 TI - Different effects of NMDA/group I metabotropic glutamate receptor agents in delta and mu-opioid receptor agonist-induced supraspinal antinociception. AB - The N-methyl-D-aspartate (NMDA) and metabotropic glutamate (mGlu) receptors are involved in nociceptive transmission in the central nervous system. The present study was designed to study the effects of NMDA and group I mGlu receptor agents on delta- and mu-opioid receptor agonist-induced antinociception in the mouse brain. Intracerebroventricular (i.c.v.) treatment with the non-competitive NMDA receptor antagonist dizocilpine and the group I mGlu receptor antagonist (S)-4 carboxyphenylglycine ((S)-4CPG) significantly attenuated the antinociception induced by the delta-opioid receptor agonists [D-Pen(2), Pen(5)]enkephalin (DPDPE), (-)-TAN 67 and [D-Ala(2)]deltorphin II. On the contrary, i.c.v. administration of dizocilpine and (S)-4CPG slightly but significantly enhanced the antinociception induced by the mu-opioid receptor agonist [D-Ala(2), N-Me Phe(4), Gly(5)-ol]enkephalin (DAMGO). Under these conditions, i.c.v. administration of NMDA and the group I mGlu receptor agonist 3,5 dihydrophenylglycine (DHPG) significantly enhanced the antinociception induced by delta-opioid receptor agonists, whereas both reduced DAMGO-induced antinociception. These findings suggest that the supraspinal antinociceptive actions of mu- and delta-opioid receptor agonists appear to be modulated differently by NMDA and group I mGlu receptors in the mouse. PMID- 10822051 TI - Endothelin ET(A) receptor antagonism attenuates the pressor effects of nicotine in rats. AB - The increased endothelin-1 levels observed after smoking may result from nicotine stimulated endothelin-1 production by endothelial cells. In this study, we investigated the effects of selective endothelin ET(A) receptors antagonist Cycle D-a-aspartyl-L-prolyl-D-isoleucyl-D-tryptophyl (JKC 301) and of endothelin ET(B) receptors antagonist N-cis-2, 6-dimethylpiperidino-carbonyl-L-gamma-methyl-leucyl D-L-m ethoxycarbonyl-tryptophanyl-norleucine (BQ 788) on the changes in mean arterial pressure, heart rate, and plasma thromboxane B(2) (the stable product of thromboxane A(2)) levels caused by increasing doses of nicotine (0.6, 2, 6, and 20 micromol/kg) in anesthetised rats. Nicotine (0.6, 2, and 6 micromol/kg) significantly increased the mean arterial pressure in control and BQ 788 pretreated rats, while only a nicotine dose of 2 micromol/kg) increased the mean arterial pressure in JKC 301-pretreated animals. There were no differences in the nicotine-induced changes in heart rate or in the increases in thromboxane B(2) levels among the groups treated with saline, JKC 301 and BQ 788. These results demonstrate that whereas the antagonism of endothelin ET(A) receptors attenuated the increase in blood pressure after nicotine injections, endothelin ET(B) receptor antagonism had no such effect. In addition, the antagonism of endothelin ET(A) or ET(B) receptors did not affect thromboxane A(2) production after nicotine administration. These findings suggest that endothelin-1 may have a role in the acute effects of nicotine. PMID- 10822052 TI - The anandamide transport inhibitor AM404 activates vanilloid receptors. AB - The possibility that the anandamide transport inhibitor N-(4-hydroxyphenyl) 5,8,11,14-eicosatetraenamide (AM404), structurally similar to the vanilloid receptor agonists anandamide and capsaicin, may also activate vanilloid receptors and cause vasodilation was examined. AM404 evoked concentration-dependent relaxations in segments of rat isolated hepatic artery contracted with phenylephrine. Relaxations were abolished in preparations pre-treated with capsaicin. The calcitonin-gene related peptide (CGRP) receptor antagonist CGRP-(8 37) also abolished relaxations. The vanilloid receptor antagonist capsazepine inhibited vasodilation by AM404 and blocked AM404-induced currents in patch-clamp experiments on Xenopus oocytes expressing the vanilloid subtype 1 receptor (VR1). In conclusion, AM404 activates native and cloned vanilloid receptors. PMID- 10822053 TI - Vasorelaxant effect of a phosphodiesterase 3 inhibitor, olprinone, on isolated human radial artery. AB - The radial artery is currently used as a viable arterial conduit for myocardial revascularization. The aims of this study were to identify phosphodiesterase 3 isoenzyme in the human radial artery isolated for coronary artery bypass grafting, and to examine the vasorelaxant effect of a cardiotonic and vasodilating phosphodiesterase 3 inhibitor, 1, 2-dihydro-6-methyl-2-oxo-5 (imidazo[1,2-a]pyridin-6-yl)-3-pyridine carbonitrile hydrochloride monohydrate (olprinone). The phosphodiesterase 3 isoenzyme was separated from the radial artery by DEAE-Sepharose chromatography. Olprinone inhibited the phosphodiesterase 3 activity with an IC(50) value of 1.25 microM. Olprinone relaxed the phenylephrine-induced contractions of endothelium-denuded arterial strips with an EC(50) value of 0. 107+/-0.029 microM (n=5). These findings indicate that the human radial artery possesses phosphodiesterase 3 isoenzyme activity and olprinone causes potent relaxation of the arterial strip in vitro through inhibition of phosphodiesterase 3 isozyme activity. PMID- 10822054 TI - 5'-Guanidinonaltrindole, a highly selective and potent kappa-opioid receptor antagonist. AB - 5'-Guanidinonaltrindole (GNTI) possesses 5-fold greater opioid antagonist potency (K(e)=0.04 nM) and an order of magnitude greater selectivity (selectivity ratios >500) than the prototypical kappa-opioid receptor antagonist, norbinaltorphimine, in smooth muscle preparations. Binding and functional studies conducted on cloned human opioid receptors expressed in Chinese hamster ovarian (CHO) cells afforded pA(2) values that were comparable to the smooth muscle data. In view of the high selectivity and potency of GNTI, it is a potentially valuable pharmacological tool for opioid research. PMID- 10822055 TI - Effect of a selective neuropeptide Y Y(2) receptor antagonist, BIIE0246 on neuropeptide Y release. AB - We have examined the selective neuropeptide Y Y(2) receptor antagonist, (S)-N(2) [[1-[2-[4-[(R,S)-5,11-dihydro-6(6h)-oxodibenz[b, e]azepin-11-yl]-1-piperazinyl]-2 oxoethyl]cyclopentyl]acetyl ]-N-[2-[1 ,2-dihydro-3,5(4H)-dioxo-1,2-diphenyl-3-H 1,2, 4-triazol-4-yl]ethyl]-argininamid (BIIE0246) on neuropeptide release from rat hypothalamic slices in vitro. BIIE0246 prevented neuropeptide Y-(13-36) induced reduction in basal and K(+)-stimulated neuropeptide Y release. Addition of BIIE0246 alone enhanced K(+)-stimulated neuropeptide release, without affecting basal release. These data are consistent with anatomical and functional studies suggesting a pre-synaptic role for neuropeptide Y Y(2) receptors in regulating rat hypothalamic neuropeptide Y release in the rat. PMID- 10822056 TI - Effect of norepinephrine release on adrenoceptors in severe seizure genetically epilepsy-prone rats. AB - The genetically epilepsy-prone rat (GEPR) seizure model is characterized by extensive abnormalities in brain noradrenergic function. Earlier studies had suggested that GEPRs might not regulate adrenoceptors in a normal fashion. The purpose of the present study was to determine if GEPR-9s are capable of up and down regulation of alpha(1)- and beta-adrenoceptors in response to increments or decrements in extracellular norepinephrine. Seizure induction has been shown to increase extracellular norepinephrine. Chronic sound or electroshock-induced seizures caused down regulation of beta-adrenoceptors in frontal cortex and in hippocampus from GEPR-9s. Similarly, chronic daily treatment with the norepinephrine reuptake inhibitor desmethylimipramine produced down regulation of beta-adrenoceptors in frontal cortex and in hippocampus from GEPR-9s. As is the case in neurologically normal animals, chronic electroshock-induced seizure did not cause down regulation of beta-adrenoceptors in 6-hydroxydopamine pretreated GEPR-9s. Chronic electroshock treatment also caused up-regulation of alpha(1) adrenoceptors in frontal cortex but not in hippocampus. In 6-hydroxydopamine pretreated GEPR-9s, chronic electroshock treatment caused a further up-regulation of alpha(1)-adrenoceptors in frontal cortex but not in hippocampus. Taken together, these results indicate that GEPR-9s are capable of up and down regulation of alpha(1)- and beta-adrenoceptors in a manner that is qualitatively similar to the regulation of these receptors in normal animals. Whether the regulation of brain adrenoceptors is quantitatively different in GEPRs from normal animals remains to be established. PMID- 10822057 TI - The pharmacology of a dopamine receptor in the locust nervous tissue. AB - A dopamine receptor in the nervous tissue of the desert locust (Schistocerca gregaria Forskal) was studied using ?3Hlysergic acid diethylamide (LSD) as the radioligand. Its expression is almost entirely restricted to the mushroom bodies, centres for learning and memory in the insect brain. This G-protein coupled receptor is present in relatively low concentrations in the locust brain (35 fmol/mg protein). The pharmacological characterisation reveals high affinity for the putative natural agonist dopamine (K(i)=28 nM). Substances with high subtype specificity for vertebrate dopamine receptors such as SCH 23390 (K(i)=639 nM) and sulpiride (K(i)=21,200 nM) have low affinity for the locust neuronal dopamine receptor. In opposite, substances with a broad pharmacological profile such as LSD, spiperone (K(i)=7.26 nM), and chlorpromazine (K(i)=9.52 nM) have high affinity properties. Comparison of the pharmacological data reveals no significant homology to any vertebrate dopamine receptor class characterised so far. This uncertainty about the pharmacological relatedness of insect dopamine receptors mirrors the available molecular data. It is almost impossible to classify cloned insect dopamine receptors into vertebrate dopamine receptor schemes. This lack of pharmacological relatedness opens the opportunity to develop highly specific insecticides against insect dopamine receptors. PMID- 10822058 TI - Activation of G protein by opioid receptors: role of receptor number and G protein concentration. AB - The collision-coupling model for receptor-G-protein interaction predicts that the rate of G-protein activation is dependent on receptor density, but not G-protein levels. C6 cells expressing mu- or delta-opioid receptors, or SH-SY5Y cells, were treated with beta-funaltrexamine (mu) or naltrindole-5'-isothiocyanate (delta) to decrease receptor number. The time course of full or partial agonist-stimulated ?35SGTPgammaS binding did not vary in C6 cell membranes containing <1-25 pmol/mg mu-opioid receptor, or 1. 4-4.3 pmol/mg delta-opioid receptor, or in SHSY5Y cells containing 0. 16-0.39 pmol/mg receptor. The association of ?35SGTPgammaS binding was faster in membranes from C6mu cells than from C6delta cells. A 10-fold reduction in functional G-protein, following pertussis toxin treatment, lowered the maximal level of ?35SGTPgammaS binding but not the association rate. These data indicate a compartmentalization of opioid receptors and G protein at the cell membrane. PMID- 10822059 TI - Antinociceptive activity of combination of morphine and NMDA receptor antagonists depends on the inter-injection interval. AB - The actual time-course of morphine antinociception is shorter than what would be predicted from its elimination kinetics, suggesting the presence of an acute tolerance phenomenon. Since antagonists acting at NMDA subtype of glutamate receptors were repeatedly shown to prolong acute morphine antinociception, acute tolerance may be attributed to hyperactivity of NMDA receptors. The ability of various site-selective NMDA receptor antagonists to affect morphine antinociception (tail-flick test) was assessed in mice 30 and 120 min after acute morphine challenge. Competitive NMDA receptor antagonist 3-(2-carboxypiperazin-4 yl)-1-propenyl-1-phosphonic acid (D-CPPene) (SDZ EAA 494; 0.1-1 mg/kg), low affinity channel blockers 1-amino-3,5-dimethyl adamantane (memantine) (1-10 mg/kg) and 1-amino-1,3,3,5,5-pentamethyl-cyclohexan hydrochloride (MRZ 2/579) (1 10 mg/kg), glycine site antagonists 5-nitro-6,7-dichloro-1, 4-dihydro-2,3 quinoxalinedione (ACEA-1021) (5 or 10 mg/kg) and 8-chloro-4-hydroxy-1-oxo-1,2 dihydropyridaliono(4, 5-b)quinoline-5-oxide choline salt (MRZ 2/576) (1-10 mg/kg) were administered intraperitoneally (i.p.) 15 or 30 min prior to the tail-flick test (i.e., interval between injections of morphine and NMDA receptor antagonist was either 0-15 or 90-105 min). ACEA-1021, MRZ 2/576 and to the lesser extent, memantine and MRZ 2/579 enhanced morphine antinociception when tests were conducted 120 but not 30 min post-morphine. D-CPPene potentiated morphine antinociception irrespective of the interval between morphine administration and the tail-flick test. The results suggest that NMDA receptor antagonists may restore analgesic activity of morphine in acutely tolerant mice. PMID- 10822060 TI - Role of the endogenous cannabinoid system in the formalin test of persistent pain in the rat. AB - It has been suggested that administration of a cannabinoid CB(1) (SR141716A ?N (piperidin-1-yl)-5-(4-chlorophenyl)-1-(2, 4-dichlorophenyl)-4-methyl-1-H-pyrazole 3-carboxamide) and CB(2) (SR144528 ?N-[(1S)-endo-1, 3, 3-trimethyl bicyclo ?2.2.1 heptan-2-yl]-5-(4-chloro-3-methylphenyl)-1-(4-methylbenzyl)-pyr azo le- 3 carboxamide?) receptor antagonists to mice potentiates inflammatory hyperalgesia by removing an endogenous cannabinoid tone. We examined whether the behavioural response to s.c. formalin injection in rats is similarly enhanced. A total of 30 animals received SR141716A (0.5 or 5 mg/kg) or SR144528 (0.3 or 3 mg/kg) 30 min before 1% formalin. Pain behaviour was quantified using the composite weighted pain score technique (CPS-WST(0,1,2)). An overall CPS-WST(0,1,2) was calculated for each phase and groups were compared (analysis of variance). The results obtained in the control group confirmed the characteristic biphasic behavioural response to formalin injection. None of antagonist groups had a significant increase in overall CPS-WST(0,1,2) compared to the control. Indeed, a significant decrease in CPS-WST(0,1,2) scores for both phases was detected in most of all of the groups, except SR141716A at 5 mg/kg. Levels of endogenous cannabinoids (anandamide, palmitoylethanolamide, 2-arachidonylglycerol) were measured from rats hind-paw skin 1 h after s.c. injection of 0.9% saline (100 microl), 1% (50 microl), 2. 5% (50 microl) and 5% (100 microl) formalin. The concentration of endocannabinoids did not differ between control and formalin-induced inflammation groups. The activity of anandamide amidohydrolase in hind-paw skin also did not change after treatment with formalin. In conclusion, cannabinoid antagonists do not enhance formalin-evoked pain behaviour. These results suggest that, in this model, endogenous cannabinoids do not tonically attenuate inflammatory hyperalgesia. PMID- 10822061 TI - Lack of synergism between caffeine and SKF 38393 on rotational behavior in 6 hydroxydopamine-denervated rats. AB - We have recently shown a synergistic effect between caffeine and the dopamine D(2) receptor agonist, bromocriptine, on contralateral rotational behavior in unilaterally 6-hydroxydopamine-denervated rats. In addition, we found that bromocriptine prevented caffeine-induced tolerance to this behavior following repeated treatment. In the present study, we investigated whether or not the dopamine D(1) receptor agonist, (+/-)-phenyl-2,3,4, 5-tetrahydro-(1H)-3 benzazepine-7,8-diol (SKF 38393), presented similar characteristics. Different groups of rats received simultaneous injections of either vehicle plus vehicle, caffeine (40 mg/kg) plus vehicle, SKF 38393 (0.5, 1, 2, and 4 mg/kg) plus vehicle, or caffeine plus SKF 38393 (0.5, 1, 2, and 4 mg/kg) for 5 consecutive days, and both ipsilateral and contralateral rotational behavior was measured. Results showed that, on the first day of treatment, caffeine produced significantly more rotational behavior than did a low dose of SKF 38393 (0.5 mg/kg), and significantly less turning than at higher doses (2 and 4 mg/kg). Combined treatment with caffeine and a high dose of SKF 38393 (4 mg/kg) produced significantly more rotational behavior than did caffeine plus vehicle. With repeated administration, caffeine produced sustained tolerance to its effects on rotational behavior, whereas SKF 38393 did not. In the groups treated with low doses of SKF 38393 (0.5, and 1 mg/kg) plus caffeine, tolerance was observed while in the groups that received high doses of SKF 38393 (2 and 4 mg/kg) plus caffeine, no tolerance was observed to rotational behavior. These results suggest that maximal stimulation of dopamine D(1) receptors may be needed to prevent the tolerance effects of caffeine in this animal model. This finding may have clinical relevance to the therapeutic treatment of Parkinson's disease. PMID- 10822062 TI - Antiparkinson potential of delta-opioid receptor agonists. AB - delta-Opioid receptors, present in very high concentrations in striatum and overlying cortex, are thought to be involved in a number of processes, including analgesia, mood, reward, modulation of neuronal excitability, and alterations in neurotransmitter release. Given the localization of the receptors in motor circuits in brain, we thought it of interest to study the antiparkinson potential of delta-opioid receptor agonists. Rats were given unilateral 6-hydroxydopamine lesions of the nigrostriatal tract, and following recovery, were tested for rotational activity. Tonazocine mesylate is a nonpeptide, partial delta-opioid receptor agonist with mu-receptor antagonist properties. Tonazocine (0.1-10 mg/kg) evoked a dose-related, ipsilateral rotation, consistent with augmentation of dopaminergic function on the unlesioned side. The rotation evoked by tonazocine was blocked by the selective delta-opioid receptor antagonist naltrindole, suggesting that the effect was mediated by delta-opioid receptors. The full delta-opioid receptor agonist (+)-4-?9-alpha-R)-alpha-(2S,5RO-4-allyl-2, 5-dimethyl-1-piperaziny l)-3-methoxybenzyl-N,N-diethylbenzamide (SNC-80) produced both contralateral and ipsilateral rotation. Tonazocine additionally augmented the effects of L-3,4 dihydroxyphenylalanine (L-DOPA) on reserpine-induced suppression of motor activity. Binding affinities and efficacies of tonazocine and SNC-80 against mu-, kappa-, and delta-opioid receptors were also confirmed and compared to standards. These data suggest therapeutic potential of agents interacting with delta-opioid receptors, and indicate some differences in the activities of tonazocine and SNC-80. PMID- 10822063 TI - Negative inotropic effect of endothelin-1 in the mouse right ventricle. AB - Effects of endothelin-1 on the contraction and cytosolic Ca(2+) concentrations (?Ca(2+)(i)) of the mouse right ventricle were investigated. Endothelin-1 (1-300 nM) elicited a negative inotropic effect in a concentration-dependent manner. The endothelin-1-induced negative inotropy was antagonized by a selective endothelin ET(A) receptor antagonist, BQ-123 (cyclo ?Asp-Pro-Val-Leu-Trp-; 3, 10 microM). Endothelin-1 reduced the peak amplitudes of both the ?Ca(2+)(i) transient and contraction without changing inward Ca(2+) current. The relationship between peak amplitude of ?Ca(2+)(i) and peak force generated by changing the extracellular Ca(2+) concentration (?Ca(2+)(o)) was not affected by endothelin-1. In addition, the trajectory of the ?Ca(2+)(i)-contraction phase plane diagram obtained at 2 mM ?Ca(2+)(o) in the absence of endothelin-1 was superimposable on that obtained at 4 mM ?Ca(2+)(o) in the presence of endothelin-1 (300 nM). Endothelin-1 (300 nM) translocated protein kinase C from cytosol to membrane, suggesting activation of protein kinase C. Further, a selective protein kinase C inhibitor, bisindolylmaleimide I (10 microM), inhibited the endothelin-1-induced negative inotropy. These results suggest that endothelin-1 elicits negative inotropy by reducing the amplitude of the ?Ca(2+)(i) transient without changing inward Ca(2+) current through the activation of the endothelin ET(A) receptor followed by protein kinase C activation in the mouse right ventricle. PMID- 10822064 TI - Inhibition of the Na(+)/H(+) exchanger attenuates phase 1b ischemic arrhythmias and reperfusion-induced ventricular fibrillation. AB - The sodium-hydrogen exchanger-isotype 1 (NHE-1) plays a critical role in myocardial ischemia-reperfusion injury. While studies employing less selective sodium-hydrogen inhibitors have demonstrated antiarrhythmic activity, only one study has examined the in vivo efficacy of selective NHE-1 inhibition in a canine model of ischemia-reperfusion-induced arrhythmia. In the present study, the antiarrhythmic activity of Benzamide, N-(aminoiminomethyl)-4-?4-(2 furanylcarbonyl)-1-piperazinyl -3-(methy lsulfonyl), methanesulfonate (BIIB 513), a novel NHE-1 inhibitor, was examined. An in vivo canine model of myocardial ischemia-reperfusion injury in which 60 min of left anterior descending coronary artery (LAD) occlusion followed by 3 h of reperfusion was employed. BIIB 513 was infused either prior to ischemia or prior to reperfusion. Arrhythmias were quantified by single lead electrocardiogram. Infarct size, determined by triphenyltetrazolium staining, was expressed as a percent of the area-at-risk. In vivo, NHE-1 inhibition did not affect phase 1a arrhythmias, which occur within the first 10 min of occlusion, however, BIIB 513 significantly reduced the incidence of ischemia-induced phase 1b arrhythmias which occur between 10 and 30 min following occlusion and the incidence of reperfusion-induced ventricular fibrillation. Furthermore, NHE-1 inhibition significantly reduced infarct size, when the drug was administered either prior to ischemia or prior to reperfusion. NHE-1 inhibition selectively reduces both ischemia-induced phase 1b arrhythmias and reperfusion-induced ventricular fibrillation, and also markedly reduces myocardial infarct size when the drug is administered prior to ischemia or prior to reperfusion. PMID- 10822065 TI - Aminoguanidine prevents concanavalin A-induced hepatitis in mice. AB - Aminoguanidine is an inhibitor of the inducible form of nitric oxide synthase (iNOS). In the present study, the effect of aminoguanidine on concanavalin A induced hepatitis was examined. Treatment of mice with concanavalin A (10 mg/kg, i.v.) induced interferon-gamma and iNOS mRNA expression in the liver before the elevation of plasma alanine aminotransferase activity. Immunohistochemical study showed the induction of iNOS protein expression in the area of necrosis. Aminoguanidine (1, 3 and 10 mg/kg, i.p.) inhibited the concanavalin A-induced elevation of alanine aminotransferase activity. Aminoguanidine (10 mg/kg, i.p.) did not inhibit concanavalin A-induced interleukin-2, interferon-gamma, tumor necrosis factor-alpha or iNOS mRNA expression in the liver. The plasma nitrite/nitrate level was elevated at 6 and 24 h after concanavalin A treatment. The elevation of nitrite/nitrate was inhibited by aminoguanidine (10 mg/kg, i.p.). From these results, we conclude that nitric oxide formed by iNOS may be involved in the development of concanavalin A-induced hepatitis. PMID- 10822066 TI - Functional evidence that gastroprotection can be induced by activation of central alpha(2B)-adrenoceptor subtypes in the rat. AB - Clonidine injected intracerebroventricularly (i.c.v.) (0.47 nmol/rat) exerted gastric mucosal protective effect against acidified ethanol. Evidence was obtained that the gastroprotective effect of clonidine was blocked by i.c.v. injected alpha(2)-adrenoceptor antagonists yohimbine (non-subtype selective antagonist), prazosin and 2-[2-(4-(O-methoxyphenyl)piperazin-1-yl)ethyl]-4,4 dimethyl-1,3-(2 H, 4H)-isoquinolindione (ARC-239) (representative alpha(2B/2C) adrenoceptor blocking agents) and opioid receptor antagonists naloxone (a non selective, moderately mu-opioid receptor preferring antagonist), naltrindole and naltriben delta-opioid receptor antagonists). The centrally injected naltrindole (0.5 nmol/rat) antagonised also the gastroprotective effect of clonidine --but not that of the delta-agonist [D-Ala(2), D-Leu(5)]enkephalin--administered peripherally. The results suggest that central alpha(2B/2C)-adrenoceptor subtypes and opioid--particularly delta--receptors are likely to be involved in the gastric mucosal protective effect of clonidine. PMID- 10822067 TI - Specific uptake of 5-hydroxytryptamine is reduced in lungs from hypoxic pulmonary hypertensive rats. AB - In this study, the aim was to determine whether 5-hydroxytryptamine (5-HT) removal by the pulmonary endothelium is reduced in 1-week hypoxic, pulmonary hypertensive rats by directly measuring [3H]5-HT uptake in isolated lungs. In lungs from hypoxic rats, specific 5-HT uptake was reduced. This was due to a 50% decrease in the maximal initial rate of uptake rather than a decrease in affinity of 5-HT for its transporter. It is possible that reduced removal of 5-HT may contribute to the elevation in plasma levels of this vasoactive amine in pulmonary hypertension. PMID- 10822068 TI - Immunomodulation by cannabinoids is absent in mice deficient for the cannabinoid CB(2) receptor. AB - Cannabinoids have immunomodulatory as well as psychoactive effects. Because the central cannabinoid receptor (cannabinoid CB(1) receptor) is highly expressed in many neuronal tissues and the peripheral cannabinoid receptor (cannabinoid CB(2) receptor) is highly expressed in immune cells, it has been suggested that the central nervous system effects of cannabinoids are mediated by cannabinoid CB(1) receptors and that the immune effects are mediated by cannabinoid CB(2) receptors. To test this hypothesis, we have generated the first mouse strain with a targeted mutation in the cannabinoid CB(2) receptor gene. Binding studies using the highly specific synthetic cannabinoid receptor agonist (-)-cis-3-?2-Hydroxy-4 (1, 1-dimethylheptyl)phenyl-trans-4-(3-hydroxypropyl)cyclohexanol (?3HCP 55,940) revealed no residual cannabinoid binding sites in the spleen of the cannabinoid CB(2) receptor knockout mice, while binding in the central nervous system was unchanged. Cannabinoid CB(2) receptor knockout mice, which appear healthy, are fertile and care for their offspring. Fluorescence activated cell sorting (FACS) analysis showed no differences in immune cell populations between cannabinoid CB(2) receptor knockout and wildtype mice. We investigated the immunomodulatory effects of cannabinoids in cannabinoid CB(2) receptor deficient mice using a T cell co-stimulation assay. Delta(9)Tetrahydrocannabinol inhibits helper T cell activation through macrophages derived from wild type, but not from knockout mice, thus indicating that this effect is mediated by the cannabinoid CB(2) receptor. In contrast, central nervous system effects of cannabinoids were not altered in these mice. Our results suggest that cannabinoid CB(2) receptor specific ligands may be clinically useful in the modulation of macrophage immune function while exhibiting no central nervous system activity. Furthermore, we conclude that the cannabinoid CB(2) receptor knockout mouse is a useful animal model in which to study the role of the cannabinoid system in immunoregulation. PMID- 10822069 TI - Sudden infant death syndrome, childhood thrombosis, and presence of genetic risk factors for thrombosis. AB - Sudden infant death syndrome or "cot death" has until the late eighties been a significant cause of death in children between the ages of 1 month and 1 year. Approximately two per 1000 children born alive dies of sudden infant death syndrome each year in Western Europe, North America, and Australia. The vulnerability of the infant brain stem to ischemia has been suggested to be a conceivable cause of sudden infant death syndrome. This is compatible with a hypothesis that genetic risk factors for cerebral thrombosis could cause microinfarction in the brain stem during the first month of life, affecting vital centers or their blood supply. The presence of three common point mutations seen in families with thrombophilia (1691G-->A in the coagulation factor V gene, 677C- >T in the methylenetetrahydrofolate reductase gene, and the 20210G-->A mutation in the prothrombin gene) could increase the risk for thrombosis in the child. This prompted us to investigate these genetic markers of thromboembolic disease in 121 cases of sudden infant death syndrome and in relevant controls, in the expectation of a more frequent occurrence of these markers if thrombosis is an etiological factor in sudden infant death syndrome. The frequency of homozygous 1691G-->A mutation in SIDS cases was higher than expected (odds ratio: 7.3, 95% confidence interval, 1.2-45.8). The allele frequencies (theta;) in cases of sudden infant death syndrome of the 1691G-->A, 677C-->T, and 20210G-->A alleles was 2.6% (1.0-5.5), 32.6% (26.8-38.9), and 0.9% (0.1-3.4), respectively. None of the allele frequencies found in the background population (3.4% for the 1691G-->A allele, 29% for the 677C-->T allele, and 1% for the 20210G-->A allele) differed significantly from that in cases of sudden infant death syndrome. In 5,251,027 inhabitants in Denmark, the incidence of venous thromboembolism was 0.9 per 1000 per year in the background population, and less than one-thousandth of these were children. Consequently it is not likely that venous thrombosis is a major cause of sudden infant death syndrome. On the other hand, this does not exclude other known or unknown risk factors for thrombosis as possible etiological factors for sudden infant death syndrome. It is likely that we must continuously employ the exclusion principle on possible etiological causes in genetic material from a large group of victims of sudden infant death syndrome if the phenomenon of sudden infant death syndrome is to be ascribed to a specific hereditary disorder. PMID- 10822070 TI - Antiphospholipid antibodies in children without and in adults with and without thrombophilia. AB - Antiphospholipid antibodies (APAs) are considered risk factors in patients with thromboembolic diseases. Although the incidence of such acquired coagulation disturbances in adults are well described, only few data exist for children. Therefore, in a first step to collect new data we analyzed the presence of different APAs in 202 consecutive children and compared them with two groups of adults. The children screened for APA were exclusively those who did not have any thromboembolic complications or a tendency for thrombophilia due to other underlying diseases such as systemic lupus or malignancy in their past or present medical history. Consecutive blood samples were evaluated from routine laboratory specimens. The two groups of adults comprised 200 patients after deep vein thrombosis and 200 patients without thromboembolic events that served as controls. Four lupus anticoagulant (LA) screening tests were determined: the dilute Russell's viper venom test; a lupus anticoagulant-sensitive activated partial thromboplastin time reagent; a second lupus-sensitive activated partial thromboplastin time; and the Kaolin clotting time. Furthermore, three different antiphospholipid antibodies ELISA assays against cardiolipin (ACA), beta2 glycoprotein I, and phosphatidyl-serine, were determined. The children had a much higher prevalence for LA than did the adults. On the other hand, their values for ACA were significantly lower than in adults with a history of thromboembolism. Findings in children were similar to the normal adult group. This has to be taken into account when evaluating children with thromboembolic diseases. PMID- 10822071 TI - Heparin, platelet aggregation, neutrophils, and cardiopulmonary bypass. AB - Cardiopulmonary bypass (CPB) is associated with both neutrophil activation and failure of platelets to form large stable aggregates. We aimed to determine the effects of heparin and of neutrophil activation on platelet aggregation in whole blood. Fourteen patients undergoing routine aortocoronary bypass grafting and NSAID-free for over 7 days were studied before and after heparinisation, and at end-CPB. Whole blood, anticoagulated with rHirudin, was stirred for 3 minutes, and macroaggregation in response to collagen (0.6 microg. mL(-1)) or the neutrophil stimulant fMLP (10(-7)M) was determined by whole blood impedance aggregometry. Microaggregation was measured by counting unaggregated single platelets (corrected for haemodilution). The blood of volunteers was studied in vitro. PATIENTS: Before CPB, heparin effectively abolished platelet macroaggregation induced by collagen (20.5 to 1.4 Ohms) or fMLP (3.9 to 0 Ohms (p<0.0001). CPB had no additional effect. Heparinisation also reduced the platelet count from 127 (110-170) to 95 (64-117). The inhibition of macroaggregation could not be reversed by ex vivo heparinase. VOLUNTEERS: In vitro, the same heparin concentration, as measured in vivo (4 micromL(-1)), inhibited collagen-induced macroaggregation (20.3 to 14.7 Omega), but this effect was less than that observed ex vivo and was reversed by heparinase. In vitro heparin promoted fMLP macroaggregation (2.9 to 8.6 Omega). The inhibition of macroaggregation resulted from heparinisation, per se, rather than CPB and was insensitive to heparinase. There was less inhibition by in vitro heparin, which was reversible by heparinase, indicating a direct effect of heparin in vitro. The disparate findings are suggestive of an indirect action by heparin in vivo on macroaggregation, although heparin had a small direct stimulatory action on microaggregation. PMID- 10822072 TI - Platelet-derived microparticles in patients with arteriosclerosis obliterans: enhancement of high shear-induced microparticle generation by cytokines. AB - We evaluated the plasma concentrations of cytokines and platelet-derived microparticles in patients with arteriosclerosis obliterans and studied the effect of cytokines on platelet-derived microparticle generation under high shear stress. Interleukin-6 levels peaked at 48 hours after vascular surgery, while thrombopoietin started to increase at 24 to 48 hours postoperatively and peaked on the seventh day. Platelet activation markers were increased in the arteriosclerosis obliterans patients preoperatively. Levels of P-selectin and CD63 both increased further, peaking at 6 to 24 hours postoperatively. Platelet derived microparticle levels were also increased preoperatively. At 6 hours postoperatively, the plasma level of platelet-derived microparticles was significantly increased. Plasma platelet-derived microparticle level was lower at 12 hours but only returned to the preoperative value at 7 days after grafting. There was a difference in the platelet-derived microparticle level at 7 days between patients with or without antiplatelet therapy (cilostazol). The effect of cytokines on platelet activation under high shear stress was also studied. Interleukin-6 and thrombopoietin enhanced both P-selectin expression and platelet derived microparticle generation under high shear stress. These results suggest that platelet-derived microparticles are released by platelet activation after vascular grafting when certain cytokines increase under high shear stress and that antiplatelet therapy may reduce platelet-derived microparticle levels postoperatively. PMID- 10822073 TI - Antithrombotic treatment (argatroban vs. heparin) in coronary angioplasty in angina pectoris: effects on inflammatory, hemostatic, and endothelium-derived parameters. AB - To evaluate the suitability of two anticoagulants (heparin vs. argatroban) as adjunctive drugs during and after elective conventional percutaneous transluminal coronary angioplasty, we compared the changes in inflammatory, hemostatic, and endothelium-derived markers in groups of patients with stable angina treated with the two drugs during percutaneous transluminal coronary angioplasty. Twenty-seven patients were randomly allocated to either group 1 (15 patients who received an empiric dose of heparin and aspirin as anticoagulant), or group 2 (12 patients who received an alternative regimen of argatroban and aspirin). Both drugs were administered as a bolus followed by continuous infusion for 96 hours during and after percutaneous transluminal coronary angioplasty. There were no differences in the inflammatory response induced by percutaneous transluminal coronary angioplasty in both groups, but the fibrinogen concentration significantly decreased during percutaneous transluminal coronary angioplasty in group 2. Decreased platelet counts and increased mean platelet volume were observed during percutaneous transluminal coronary angioplasty in both groups. The levels of prothrombin fragment 1+2 and thrombin antithrombin III complex increased markedly during percutaneous transluminal coronary angioplasty. Group 2 showed a more rapid return to the baseline levels of these two markers than group 1. Secondary fibrinolysis was evidenced by a steep increase of D-dimer after percutaneous transluminal coronary angioplasty in both groups. In contrast to the significant decrease in antithrombin activities during percutaneous transluminal coronary angioplasty in group 1, no marked change in these markers was found in group 2. Although the levels of von Willebrand factor and plasminogen activator inhibitor 1 showed essentially the same changes in both groups during and after percutaneous transluminal coronary angioplasty, more markedly increased levels of tissue type plasminogen activator and type plasminogen activator-plasminogen activator inhibitor-1 complex after percutaneous transluminal coronary angioplasty were found in group 1 than in group 2. While neither drug had any effect on the percutaneous transluminal coronary angioplasty-induced inflammatory response, argatroban may more effectively inhibit the generated thrombin and prevent antithrombin consumption during and after percutaneous transluminal coronary angioplasty. PMID- 10822074 TI - Platelet glycoprotein Ia gene dimorphism alpha2-807 in malignant arrhythmia in coronary artery disease. AB - It has been shown recently that the variable expression of the platelet collagen receptor integrin alpha2beta1 predisposes to thrombotic risk on the one hand and hemorrhagic risk on the other hand. The level of expression of the integrin alpha2beta1 is genetically controlled and associated with the alpha2-807 dimorphism. The expression level of this platelet collagen receptor may play a central role in the rapidly evolving coronary artery lesions that lead to malignant arrhythmia and sudden cardiac death. We studied allelic frequencies of the alpha2-807 dimorphism for their relation as a risk factor for malignant arrhythmia in a well-defined subgroup of patients with coronary artery disease. We compared allelic frequencies (by sequence specific primer polymerase chain reaction) of the dimorphism that is associated with integrin alpha2beta1 levels in 94 Caucasoid survivors of sudden cardiac death with a matched group of 106 patients with coronary artery disease without sudden cardiac death. Gene frequencies in the patient groups did not differ and were similar to those in the general population represented by 217 healthy individuals. There was no overrepresentation of an allele in any group. The inherited dimorphism that is associated with the levels of platelet integrin alpha2beta1 is not associated with malignant arrhythmia in coronary artery disease patients. PMID- 10822075 TI - Clinical outcome of self-management of oral anticoagulation in patients with atrial fibrillation or deep vein thrombosis. PMID- 10822076 TI - Platelet function analysis with PFA-100 in patients medicated with acetylsalicylic acid strongly depends on concentration of sodium citrate used for anticoagulation of blood sample. AB - Our data from 214 patients after cessation of long-term therapy with 100 mg/d ASA demonstrate that the determination of platelet-related primary hemostasis in citrated whole blood with PFA-100 is a reliable and sensitive method for the detection of ASA-induced platelet dysfunction. However, the sensitivity of the method is strongly dependent on concentration of sodium citrate used as anticoagulant. The results of PFA-100 testing show a clearly enhanced sensitivity for ASA when blood samples were collected with 0.129 M rather than 0.106 M sodium citrate. According to sample stability, PFA-100 results can only be confirmed up to 1 hour postcollection when blood was anticoagulated with 0.129 M but not with 0.106 M sodium citrate. Therefore, we recommend that testing with PFA-100 in patients with suspected ASA-induced platelet dysfunction should be performed exclusively in blood collected in buffered 0.129 M sodium citrate. PMID- 10822077 TI - Cloning and expression of the platelet-specific collagen receptor glycoprotein VI. AB - Platelet glycoprotein VI (GP VI) was purified from platelet membranes and its internal amino acid sequences were determined. The cloned cDNA of GP VI indicates an open reading frame coding for 20 amino acid signal sequences and a mature protein of 319 amino acids. Its extracellular region has two Ig-like domains and a mucin-like, Ser/Thr-rich region, suggesting that GP VI is a member of the paired Ig-like receptor family. GP VI-transfected cells contained convulxin (reactive) and antibody against recombinant GP VI-reactive protein bands that migrated at the same position as platelet GP VI in SDS/PAGE-electroblotting. These data indicate that the protein deduced from the cloned cDNA corresponds to platelet GP VI. PMID- 10822078 TI - Effects of binding of ligand (FVIIa) to induced tissue factor in human endothelial cells. AB - The tissue factor protein is structurally related to the cytokine receptors and ligand binding (factor VIIa) has been reported to give an intracellular calcium signal, thus indicating that tissue factor is a true receptor. In view of the attempts to use recombinant factor VIIa as a therapeutic agent in hemophilia, its binding effects may be of clinical interest. We have studied the effect of ligand binding to human endothelial cells that were stimulated with interleukin-1 to express tissue factor. Human umbilical cord vein endothelial cells produce and release a wide variety of proteins that participate in coagulation and fibrinolysis, and we have investigated whether binding of recombinant factor VIIa to tissue factor altered the release of some of these compounds. Three main findings are reported. (1) After an initial increase, the measurable tissue factor activity in endothelial cells decreased more rapidly in the presence of factor VIIa (half-life 3.7+/-0.7 hours) than in its absence (half-life 7.4+/-1.5 hours). This difference was not seen when tissue factor antigen was measured, indicating that ligand binding did not increase the degradation of the protein. (2) Tissue factor pathway inhibitor was detected on the cell surface, in cell homogenates, and in cell medium. When recombinant factor VIIa was added to the cells there was a significant decrease in the release of tissue factor pathway inhibitor to the medium. Four hours after recombinant factor VIIa was added, the levels were 7.5-fold higher in the medium of untreated cells compared to the medium of cells treated with recombinant factor VIIa. (3) We observed increased release of von Willebrand factor (vWF). After 1 and 6 hours with recombinant FVIIa the release was significantly greater than in controls without FVIIa. We did not detect significant differences in the release of tissue plasminogen activator or tissue factor pathway inhibitor. PMID- 10822079 TI - Proteolysis of subendothelial adhesive glycoproteins (fibronectin, thrombospondin, and von Willebrand factor) by plasmin, leukocyte cathepsin G, and elastase. AB - The degradation of extracellular matrix (ECM) adhesive glycoproteins, fibronectin (FN), thrombospondin (TSP) and von Willebrand factor (vWF), by human leukocyte cathepsin G and elastase, and by plasmin or thrombin, was analysed by immunoblotting after incubation of physiologic doses of the proteases with confluent human umbilical vein endothelial cells. Elastase induced an almost complete disappearance of intact FN, TSP, and vWF from the ECM at 0.02 units/ml within 5 minutes of incubation at 37 degrees C. Plasmin (0.2 units/ml) was also active on all three substrates, whereas cathepsin G (0.2 units/ml) had a preferential effect on TSP. Most remarkably, these degradations occurred with no apparent change in endothelial cell morphology, as shown by phase-contrast microscopy. In contrast, thrombin (0.2 units/ml) had no apparent proteolytic action on ECM glycoproteins, where it induced cell retraction and rounding. The release of adhesive glycoproteins from the ECM was accompanied by the detection of proteolytic fragments in the conditioned medium. Kinetic studies indicated that proteolysis started within minutes and proceeded for at least 1 hour. TSP was extremely sensitive to degradation by all enzymes except thrombin, whereas vWF released from the ECM was more resistant to proteolysis than constitutively secreted vWF, and FN was poorly degraded by plasmin. Our results indicate that serine proteinases, locally produced during inflammation and/or thrombolysis, can release extracellular matrix components and generate proteolytic fragments with potential biological activities. PMID- 10822080 TI - An inhibitor of activated thrombin-activatable fibrinolysis inhibitor potentiates tissue-type plasminogen activator-induced thrombolysis in a rabbit jugular vein thrombolysis model. AB - When activated in vitro, thrombin-activatable fibrinolysis inhibitor (TAFI) slows clot lysis by cleaving the C-terminal lysine and arginine residues from partially degraded fibrin. An inhibitor of carboxypeptidase isolated from potato (CPI) reverses prolongation of clot lysis by inhibiting activated TAFI. We investigated in vivo effect of TAFI inhibition on tissue-type plasminogen activator (t-PA) induced clot lysis using CPI in a rabbit jugular vein thrombolysis model. It was found necessary to further purify the CPI preparations from commercial sources by HPLC chromatography to remove endotoxin and anti-plasmin activity that would affect the endogenous fibrinolytic system. The effect of intravenous administration of the purified CPI with t-PA was determined by measuring thrombus weight at the end of 90 minutes in six groups of animals. In the control group receiving saline, the median thrombus weight was 116 mg. In the group that received CPI only (0.5 mg/kg bolus injection followed by 0.3 mg/kg/h infusion), the median thrombus weight was 121 mg. In the group that received t-PA at a dose of 10 microg/kg bolus followed by 67 microg/kg/h infusion, the median thrombus weight decreased to 86 mg. When CPI was coadministered with the same regimen of t PA, the median value further decreased to 58 mg. When animals were given three times higher the dose of t-PA (30 microg/kg bolus followed by 200 microg/kg/h infusion) in the absence or presence of CPI, median thrombus weights were 56 mg and 0 mg, respectively. Our results demonstrate that systemic coadministration of the purified CPI improves clot lysis induced by t-PA. PMID- 10822081 TI - Unexpected molecular mimicry among peptides MHC class II, blood-clotting factor X, and HIV-1 envelope glycoprotein GP120. PMID- 10822082 TI - Determining intramolecular binding sites on surface-bound von Willebrand factor under aqueous conditions. AB - The results described in this report demonstrate the feasibility of using AFM in combination with immuno-gold labeling to investigate the accessibility of various binding sites on vWF and to localize the binding site within a vWF multimer. With the aid of monoclonal antibodies [5, 11 and 23] it should be possible to use this approach to perform a quantitative assessment of the differential accessibility of various binding sites on vWF. This should allow localization and quantification of binding sites within the observed tertiary structure of the vWF, providing a measure of the accessibility, a point of reference with which the tertiary structure of vWF could be correlated to the primary sequence, and a means to determine the structural features of the antibody binding regions under physiologic buffer conditions. There are a number of obvious questions that are not addressed here: The role of different biologic and artificial surfaces; time dependent effects; vWF orientation with respect to different thrombogenic surfaces; and the location of critical binding sites, such as for platelet GPIalpha and GPIIb-IIIa binding regions in the hydrated tertiary structure of vWF. Nevertheless, the work described in this report provides essential groundwork that should provide a novel basis on which to explore the molecular steps, both structural and functional, of vWF in thrombus development. In a wider sense, this experimental approach is applicable to structure-function studies on a wide variety of proteins in physiologic environments. PMID- 10822083 TI - A new model of allergic rhinitis in rats by topical sensitization and evaluation of H(1)-receptor antagonists. AB - An animal model of chronic allergic rhinitis was developed by repeated local booster sensitization into the nasal cavity in sensitized rats. The severity of allergic rhinitis was assessed by determining the extent of two markers of nasal allergic symptoms (sneezing and nasal rubbing) after antigen challenge. The number of incidents of sneezing and nasal rubbing was markedly increased during intranasal instillation of antigen in sensitized rats. The PCA titers were also markedly elevated by intranasal sensitization. Some histamine H(1)-receptor antagonists such as chlorpheniramine, ketotifen, astemizole and epinastine inhibited the increase in antigen-induced nasal symptoms in a dose-related manner. Nasal rubbing was more potently inhibited by H(1)-receptor antagonists than sneezing. In conclusion, we developed a chronic allergic rhinitis model showing nasal symptoms in rats, and this model may be useful for evaluating the effects of drugs on allergic rhinitis. PMID- 10822084 TI - Suppression of experimental autoimmune tubulointerstitial nephritis in BALB/c mice by berberine. AB - Berberine (BB) is a protoberberine alkaloid derived from various representatives of the Berberidaceae family. Although used as a therapeutic agent, it has not been applied in the treatment of immune-mediated disorders. In the present study, BB was administered at a daily dose of 10 mg/kg for 3 consecutive days before the induction of tubulointerstitial nephritis (TIN) by injection of bovine tubular basement membrane (TBM) antigen in BALB/c mice. The animals were investigated 2 months after TBM inoculation. The intensity of pathological injuries in animals with TIN+BB decreased significantly, an effect that correlated with the improvement of renal function. Flow cytometric analysis of peripheral blood cells showed that BB caused a decrease in the number of CD3(+), CD4(+), CD8(+), and sIg(+) lymphocytes in comparison with TIN mice. The same tendency was noticed in the lymphocytes from kidney infiltrates of treated animals. The control animals treated only with BB showed a decrease in the number of CD3(+), CD4(+), CD8(+) T lymphocytes in comparison with control nontreated mice. Our results, thus, indicate that BB has an immunosuppressive effect in the TIN model, which is an analogue of various human kidney autoimmune diseases. PMID- 10822086 TI - Effects of trovafloxacin on the IL-1-dependent activation of E-selectin in human endothelial cells in vitro. AB - E-selectin is an endothelial-specific surface protein, which is transiently expressed in response to inflammatory cytokines and plays an important role in the recruitment of leukocytes to the site of infection. The effect of two fluoroquinolones, ciprofloxacin (cipro) and trovafloxacin (trova), on the interleukin-1 (IL-1)-dependent activation of E-Selectin was studied on human umbilical vein endothelial cells (HUVEC) in vitro. Trova, at 80 microg/ml, affected the transient expression of E-selectin mRNA after pro-inflammatory stimulation with IL-1 leading to a sustained expression over 24 h. Surface expression of E-selectin remained upregulated after 24 h in a higher percentage of cells when they were activated in the presence of trova, as determined by flow cytometry analysis. Moreover, the concentration of shedded soluble E-selectin (sE selectin) in the cell supernatant increased by 3.5 fold compared to those stimulated in the presence of cipro or without fluoroquinolones. Analogously, the antiproliferative effect of trova on endothelial cells was found to be more pronounced compared to cipro leading to an accumulation of cells arrested in G1 phase. These data provide evidence that accumulation of high concentration of trova in vivo in inflamed tissue might alter inflammatory responses. PMID- 10822085 TI - Mixed effects of TGF-beta on human airway epithelial-cell chemokine responses. AB - We investigated chemokine responses of human airway epithelial cells to transforming growth factor (TGF)-beta alone and in combination with tumor necrosis factor (TNF)-alpha or interferon (IFN)-gamma. TGF-beta selectively induced production of granulocyte-macrophage colony stimulating factor (GM-CSF) without significant coordinate expression of IL-8 or RANTES. TNF-alpha induced expression of both IL-8 and GM-CSF, without detectable production of RANTES. TGF beta synergistically enhanced GM-CSF production with TNF-alpha, but suppressed production and release of IL-8. IFN-gamma induced RANTES production and release; TGF-beta synergistically enhanced RANTES release induced by IFN-gamma, but had no effect on RANTES mRNA production. Taken together, these data demonstrate that TGF beta may play a pivotal role in the responsiveness of airway epithelial cells to chemotactic cytokines, by selectively enhancing GM-CSF and RANTES production while suppressing IL-8 production. This profile of chemokine responses promoted by TGF-beta would favor eosinophil, lymphocyte and monocyte recruitment, hallmarks of chronic and allergic inflammation, over neutrophil sequestration. PMID- 10822087 TI - Protective effect of a traditional Japanese medicine, Bu-zhong-yi-qi-tang (Japanese name: Hochu-ekki-to), on the restraint stress-induced susceptibility against Listeria monocytogenes. AB - In this study, the effect of traditional Japanese (Chinese) medicine, Bu-zhong-yi qi-tang (Japanese name: Hochu-ekki-to, HOT), on the restraint stress treatment (RST)-induced susceptibility against Listeria monocytogenes (L. monocytogenes) was examined. When RST was performed every day for 10 h from the day of infection, the bacterial numbers were increased at 3 and 5 days after the infection. Oral pretreatment with HOT for 7 days prevented such increases. Pretreatment with HOT prevented the suppression of antigen-specific IFN-gamma production by RST. HOT also prevented suppression of macrophage accumulation, including MHC-class II positives, in the peritoneal cavity and their bactericidal activity by RST. HOT suppressed the serum corticosterone level elevated by RST in infected mice. Taken together, the suppression of corticosterone using HOT participates in the prevention of suppressions of the bactericidal activity of macrophages, migration of macrophages and antigen-specific IFN-gamma production of Th1 cells by RST. Our findings suggest that HOT is a useful drug for patients suffering from stress disease to reduce the susceptibility to bacterial infection. PMID- 10822088 TI - Nitric oxide and its role in the induction of kinin B(1)-receptors after heat stress in the rat. AB - Kinin B(1)-receptors are inducible-receptors. They are absent under basal conditions but expressed following pathophysiological stresses. This study was designed to examine a possible role of nitric oxide (NO) in the mechanism underlying B(1)-receptor induction after heat stress (HS). Rats were divided into six groups, subjected or not to HS (42 degrees C internal temperature, 20 min) without or with treatment with nitro-L-arginine-methylester (L-NAME), a nonselective inhibitor of NO synthase (NOS) isoforms, or L-N(6)-(1-imino ethyl)lysine (L-NIL), a selective inhibitor of the inducible NOS. Twenty-four hours after HS, rats were injected with bradykinin and [des-Arg(9)]-bradykinin and hypotensive responses were recorded. In six additional groups, B(1)-receptor mRNAs were detected in aorta 5 h after HS or sham treatment. Bradykinin, a B(2) receptor agonist, induced a hypotension of a similar magnitude in all the groups studied. [des-Arg(9)]-bradykinin, a B(1)-receptor agonist, induced no response in sham rats. In rats previously subjected to hyperthermia, this agonist induced a hypotensive response, which was, respectively, decreased and increased by pretreatment with L-NAME and with L-NIL prior to hyperthermia. RT-PCR results confirmed these in vivo observations. In conclusion, this study suggests a role for NO in B(1)-receptor induction after HS as well as a possible interaction between NOS isoforms. PMID- 10822089 TI - K(+) channel-blocking alkoxypsoralens inhibit the immune response of encephalitogenic T line cells and lymphocytes from Lewis rats challenged for experimental autoimmune encephalomyelitis. AB - Alkoxypsoralens, known as DNA photomodifying agents, have been shown to block voltage-dependent K(+) channels (Kv) as well as to alleviate functional deficits in certain multiple sclerosis (MS) patients in a manner similar to 4 aminopyridine. Since Kv channel blockers are known to inhibit T cell-mediated immune responses both in vitro and in vivo, we investigated the effects of three alkoxypsoralens, 5-methoxypsoralen (5-MOP), 8-methoxypsoralen (8-MOP), and 5,8 diethoxypsoralen (H37), on the following parameters: (1) whole-cell K(+) currents of encephalitogenic, myelin basic protein-specific memory T cell line cells (MBP TCLC) derived from Lewis rats as measured by patch-clamp technique, (2) proliferation of MBP-TCLC and lymph node cells (LNC) from Lewis rats challenged for experimental autoimmune encephalomyelitis (EAE) by immunisation with spinal cord homogenate as measured by 3H-thymidine incorporation, (3) interferon-gamma (IFN-gamma) secretion of MBP-TCLC as measured by ELISA, and (4) IFN-gamma gene expression of LNC as measured by quantitative reverse transcription polymerase chain reaction (RT-PCR) with ELISA-detection. The examined alkoxypsoralens exhibited suppressive effects on the measured parameters with the same sequence of efficacy: H37>5-MOP>8-MOP. We, therefore, conclude that Kv channel-blocking alkoxypsoralens interfere with voltage-controlled signal transduction in lymphocytes and might thereby suppress immune responses in autoimmune diseases of the central nervous system and most likely also in other autoimmune disorders. Thus, alkoxypsoralens, especially the non-phototoxic substance H37, are new candidates for further studies on K(+) channel blocking immunosuppressive drugs. The agents may exert a dual beneficial effect on demyelinating diseases like MS, because they could attenuate the inflammatory process and improve axonal conductivity. PMID- 10822090 TI - Modulation of tumor necrosis factor-alpha production with anti-hypertensive drugs. AB - It is well known that some anti-hypertensive drugs affect insulin sensitivity and that tumor necrosis factor-alpha (TNF-alpha) is a mediator of obesity-associated insulin resistance. In this study, we have investigated the effect of anti hypertensive drugs, calcium (Ca) channel blockers (amlodipine, manidipine and nicardipine), an alpha(1)-blocker (doxazosin), a beta(1)-blocker (metoprolol), and a thiazide diuretic (hydrochlorothiazide), on lipopolysaccharide (LPS) induced TNF-alpha production. TNF-alpha production, measured with a bioassay and an immunoassay, was evaluated both in vivo and in vitro, by utilizing mice and a human peripheral blood mononuclear cell culture, respectively. Nicardipine, or amlodipine, manidipine and doxazosin significantly inhibited TNF-alpha production in mice at doses more than one or ten times higher than those used clinically, respectively. On the other hand, metoprolol increased TNF-alpha production at doses of more than 10 times those used clinically, whereas hydrochlorothiazide did not alter production of the cytokine. The in vivo effects of these drugs were not necessary parallel to the in vitro effects. Because high doses of these drugs in mice correspond to clinical doses and effects in human, these actions may be related to beneficial and/or harmful effects of these drugs on TNF-alpha mediated diseases, including insulin resistance. PMID- 10822091 TI - Evidence that FTY720 induces T cell apoptosis in vivo. AB - The immunosuppressant FTY720 induces a drastic decrease in blood lymphocytes, especially T cells; a decrease which is assumed to be the immunosuppressive mechanism of this drug. FTY720 causes cell death in vitro in lymphocytes and leukemia cells. However, the deletion mechanism of blood lymphocytes in vivo remains unclear. We investigated whether administration of FTY720 induced lymphocyte apoptosis in blood circulation. A marked decrease in the number of blood lymphocytes was observed within an hour after a single oral administration of FTY720 at doses of 5-10 mg/kg in rats and mice. Experiments using fluorescein isothiocyanate (FITC)-Annexin V and APO-BRDU methods revealed that FTY720 induced blood lymphocyte apoptosis in a dose-dependent manner. On the other hand, lymphocyte homing to Peyer's patches was proposed as the mechanism underlying the blood lymphocyte decrease at these doses. However, similar results were obtained when using aly/aly mice, which lack Peyer's patches and lymph nodes. Thus, we concluded that apoptosis of blood lymphocytes was induced immediately after administration of FTY720, and the cells could be immediately scavenged by phagocytes or the reticuloendothelial system in addition to Peyer's patches homing. We also concluded that T cells were highly sensitive to FTY720, which induced apoptosis in vivo. PMID- 10822093 TI - Educational links between psychiatry and family practice. PMID- 10822092 TI - Inhibition of calcineurin activity and protection against cyclosporine A induced cytotoxicity by prednisolone sodium succinate in human peripheral mononuclear cells. AB - We have investigated the effects of prednisolone sodium succinate (Pss) and cyclosporin A (CSA), applied alone or concurrently, on the release of arachidonic acid (AA) (cytosolic phospholipase A(2) (cPLA(2)) activity) and on the calcineurin (CN) activity of human peripheral blood mononuclear cells (PBMC). The cytotoxic damage to the cells treated by the drugs was estimated by the release of lactate dehydrogenase (LDH). We found that Pss (10(-5) M) could inhibit the CN activity and higher concentrations (10(-4) M) could decrease the cytotoxic damage caused by CSA (10(-4) M) during their combined application. CSA had no specific effect on the release of AA from the cells. In the combined clinical use of glucocorticosteroids (GCS) and CSA, their additive inhibitory effect on CN activity and the protective membrane influence of GCS against the cytotoxicity of CSA may be beneficial. PMID- 10822094 TI - Designing and implementing a primary care intervention trial to improve the quality and outcome of care for major depression. AB - Complex interventions, which have been shown to improve primary care depression outcomes, are difficult to disseminate to routine practice settings. To address this problem, we developed a brief intervention to train primary care physicians and nurses employed by the practice to improve the detection and management of major depression. Before recruitment began, the research team conducted academic detailing conference calls with primary care physicians and nurses, and provided in-person training with nurses and administrative staff. Administrative staff screened over 11,000 patients before their visits to identify those with probable major depression. Primary care physicians delegated increased responsibility to office nurses, who educated over 90% of patients about effective depression treatment and systematically monitored their progress over time. Early results demonstrate that community primary care practices can rebundle traditional team roles over the short-term to provide more systematic mental health treatment without adding additional personnel. A rigorous evaluation of this effort will reduce time-consuming, expensive, and often unsuccessful efforts to "translate" research intervention findings into everyday practice. PMID- 10822095 TI - Low-intensity treatment of depression in primary care: is it problematic? AB - The aim of this study was to examine patterns of care and outcomes of depressed patients under primary care during acute phase treatment. A cohort of depressed patients was assessed 6-8 weeks after starting pharmacotherapy in four large primary care clinics in a health maintenance organization. These patients (n = 1671) were receiving antidepressant treatment for a new episode of depression. To calculate main outcome measures, Structured Clinical Interview for Depression evaluated prior history and current depression status. Visit and pharmacy refill data described use of health services and antidepressant medication. Six to eight weeks after starting antidepressant therapy, 33.2% of patients had 0-3 depressive symptoms and no prior history of depression, an additional 42.3% also reported 0 3 symptoms but were at high risk of relapse, and 24. 5% were persistently depressed with 4 or more depressive symptoms. In the initial 6 weeks of treatment, these three groups showed similar use of antidepressant medication and health services. About 50% in each group had no follow-up visit for depression and 32%-42% had not refilled their antidepressant prescription. In general, depressed patients under primary care obtained low-intensity pharmacotherapy and inconsistent follow-up visits during initial acute phase treatment. Six weeks after starting antidepressant medicine, many were still symptomatic or recovered but had a high risk of depression relapse. Patients with unfavorable outcomes did not receive more intensive management than the one-third who had favorable outcomes. PMID- 10822096 TI - Psychiatrists' views of managed care and the future of psychiatry. AB - Managed care aims to insure the health of a population rather than that of an individual. This paper compiles opinions of psychiatrists and others on managed care and lists ways managed care potentially affects psychiatry. Managed care reverses the economic incentives indemnity insurance gave doctors to prolong treatment. It encourages psychiatrists to spend less time on empathic discussion and to use more standardized, less costly treatments. Many psychiatrists feel distressed about how managed care has changed their practices. Capitation care will change it further. Current trends suggest the U. S. will use and train fewer psychiatrists. Psychiatrists will spend less time with individual patients and more time planning and guiding the treatment of severely impaired patients. Many more psychiatrists will likely have unprecedented changes imposed on their careers. PMID- 10822097 TI - Second-generation antipsychotics in the emergency care setting. A prospective naturalistic study. AB - The objective of this subject was to examine the impact of the replacement of standard neuroleptics with atypical antipsychotic agents in an intensive psychiatric care unit. A mirror-image study was conducted. Cases admitted in the first semester of the year (when most of patients were treated with standard neuroleptics) were compared to cases admitted in the second semester of the year, when atypical antipsychotic agents were routinely utilized as first line treatment of patients with psychotic signs. Cases admitted in the first semester received a significantly higher daily dosage of antipsychotic drugs and more frequently received anticholinergics. In the second semester, a significantly higher number of patients received anticonvulsants, in particular valproate and gabapentin. There was no significant difference between the two groups of cases in the number of patients treated with antipsychotics, benzodiazepines, lithium, and carbamazepine and in the mean daily dose of benzodiazepines, lithium, carbamazepine, or valproate on the first day of hospitalization, the day of evaluation, and on discharge. On discharge, similar percentages of patients went home, were transferred to other Psychiatric Intensive Care Units (PICUs) or to private clinics, or left our PICU against medical advice. The length of hospitalization was similar in the two groups. There was no significant difference in the rate of aggressive or violent behavior registered in the two groups of cases. The risk of increasing violence rates, lengthening hospitalization, and facilitating patients' non-compliance should not be major concerns for physicians prescribing second-generation antipsychotics in the emergency care setting. Since these drugs have been shown to have at least similar efficacy (or greater efficacy in the case of clozapine) in the treatment of psychotic disorders as typical neuroleptics and to have a better side-effects profile, they should become first line treatment for patients with psychotic signs admitted to emergency care psychiatric facilities. PMID- 10822098 TI - Self-destructive acts occurring during medical general hospitalization. AB - Although several articles about suicide in general hospitals have been published, the rates of self-destructive individuals among various diseases and departments have not been reported previously. Moreover, self-destructive acts in Chinese general hospital inpatients have been neglected. We retrospectively investigated self-destructive incidents among medical general inpatients. A total of 75 self destructive incidents, including 15 fatalities, were identified during the 10 year study period. The self-destructive rate was 8.7 per 100,000, and the fatality rate was 1.8 per 100,000 admissions. The highest self-destructive rate occurred in patients admitted to the rehabilitation ward (33.4 per 100,000) followed by the neurology ward (29.9 per 100,000). The highest fatality rate occurred among patients in the neurology ward (6.7 per 100,000). The majority of self-destructive patients suffered from a chronic or terminal illness with the most frequent types of illnesses being malignant neoplasm (31.1%), neurological disease (20.3%), and chronic obstructive pulmonary disease (COPD, 10.8%). COPD patients had the highest rate of self-destructive behavior (64.0 per 100,000) and the highest fatality rate (16.0 per 100,000) due to these incidents. The most common self-destructive incident was knife-cutting. More than one-half (53.4%) of the self-destructive incidents occurred within the first 2 weeks of admission, and nearly one-half (46.7%) occurred during the night shift. Moreover, the majority of severe or fatal incidents also occurred during the night shift. The results suggest that close supervision of high-risk patients should be mandatory within the first 2 weeks following admission, especially during the night shift. PMID- 10822099 TI - Behavioral effect of mouse fibrinopeptide A on mouse forced swimming. AB - A specific dopamine 2 receptor antagonist, (-)sulpiride, induced an anti depressive behavior, climbing, in mice forced to swim for 6 h after the injection. The effective fraction was divided from the mouse serum using an ion exchanger and an ultra filtration method. This fraction contained fibrinopeptide A. A peptide synthesized according to the primary 6-amino acid sequence (TDTEDK) of fibrinopeptide A also remarkably increased the behavior. The present findings clearly indicate that a peptide with TDTEDK showed anti-depressive activity. PMID- 10822100 TI - Induction of high level of specific antibody response to the neutralizing epitope ELDKWA on HIV-1 gp41 by peptide-vaccine. AB - The monoclonal antibody 2F5 recognizing the neutralizing epitope ELDKWA on the C domain could neutralize 90% of the investigated HIV-1 isolates. Low levels of ELDKWA-epitope-specific antibodies were observed in HIV-1-infected individuals. To induce high levels of antibodies to ELDKW-epitope, C-domain peptide (P2) was conjugated with a carrier peptide (KGGG)(7)-K (K/G). P2-K/G-conjugate induced high level of antibodies in mice by titer 1:25,600 to ELDKWA-epitope. P2-K/G-BSA conjugate induced antibody response to ELDKWA-epitope (1:320-6400) in mice. The ELDKWA-epitope-specific antibodies of 19.8 and 34.6 microg/per milliliter serum were isolated from two rabbit antiserums (1:25,600). The levels of ELDKWA-epitope specific antibodies induced in rabbits were greater than 1 microg/ml, a level considered to confer long-term protection. These results demonstrate the potential role of the C-domain peptide of gp41 to develop an effective ELDKWA based epitope/peptide-vaccine against HIV-1. PMID- 10822101 TI - Purification and characterization of antimicrobial peptides from the skin of the North American green frog Rana clamitans. AB - Ten peptides with differential growth-inhibitory activity against the gram positive bacterium, Staphylococcus aureus, the gram-negative bacterium, Escherichia coli, and the yeast Candida albicans were isolated from an extract of the skin of a North American frog, the green frog Rana clamitans. Ranatuerin-1C (SMLSVLKNLGKVGLGLVACKINKQC), ranalexin-1Ca (FLGGLMKAFPALICAVTKKC), ranalexin-1Cb (FLGGLMKAFPAIICAVTKKC), ranatuerin-2Ca (GLFLDTLKGAAKDVAGKLLEGLKCKIAGC KP), and ranatuerin-2Cb (GLFLDTLKGLAGKLLQGLKCIKAGCKP), are members of three previously characterized families of antimicrobial peptides, first identified in the North American bullfrog Rana catesbeiana. In addition, five structurally related peptides (temporin-1Ca, -1Cb, -1Cc, -1Cd, and -1Ce), comprising 13 amino acid residues and containing a C-terminally alpha-amidated residue, belong to the temporin family first identified in the European common frog Rana temporaria. Peptides belonging to the brevinin-1, brevinin-2, esculentin-1, and esculentin-2 families, previously isolated from the skins of Asian and European Ranid frogs, were not identified in the extract. The data support the hypothesis that the distribution and amino acid sequences of the skin antimicrobial peptides are valuable tools in the identification and classification of Ranid frogs. PMID- 10822102 TI - Primary structure of CHH/MIH/GIH-like peptides in sinus gland extracts from Penaeus vannamei. AB - Peptides belonging to the CHH/MIH/GIH-family of crustacean hormones were isolated from acetic acid extracts of sinus glands isolated from eyestalks of the shrimp, Penaeus vannamei. The peptides were isolated by chromatography and molecular weights determined by MALDI mass spectrometry. Peptides in the range of 7-9 kDa and containing three disulfide bridges were selected for amino acid sequence analysis. Three peptides with the requisite properties were present in sufficient amounts for sequence analysis. Two peptides had unique sequences similar to CHH/MIH/GIH peptides from other crustaceans. A third peptide seemed to be a truncated form of one of the previous sequences. PMID- 10822103 TI - A peptide derived from pICln induced a strong hypotonic resistance in Escherichia coli cells. AB - We demonstrated previously that expression of rat pICln in Escherichia coli conferred a strong resistance to hypotonic stress. To define the intramolecular functional domain responsible for the resistance, molecular dissection of pICln was performed and the obtained peptides were expressed in E. coli. The cells expressing the peptides were exposed to a hypotonic solution, and their 'survival rates' were observed. The cells expressing only the peptides including the second acidic domain of pICln exhibited significantly higher 'survival rates' after hypotonic stress. The functional domain against hypotonicity was finally narrowed down to a peptide consisting of a 46-amino acid residue, P107-152. We conclude that the expression of P107-152 in E. coli cells could enhance their resistance to a hypotonic environment. PMID- 10822104 TI - Bioavailability of Ziconotide in brain: influx from blood, stability, and diffusion. AB - Ziconotide is a selective peptide antagonist of the N-type calcium channel currently in clinical trials for analgesia. Ziconotide reached a maximal brain concentration of between 0.003 and 0.006% of the injected material per gram of tissue at 3-20 min after i.v. injection, and this decayed to below 0.001%/g after 2 h. The structurally distinct conopeptide SNX-185 (synthetic TVIA) was considerably more persistent in brain after i.v. administration, with 0.0035% of the injected material present at 2-4 h after i.v. injection, and 0.0015% present at 24 h. Similar results (i.e. greater persistence of SNX-185) were obtained when the peptides were perfused through in vivo dialysis probes implanted into the hippocampus. Image analysis and serial sectioning showed that diffusion of Ziconotide in the extracellular fluid around the dialysis probe was minimal, with the peptide located within 1 mm of the probe after 2 h. In vitro diffusion through cultured bovine brain microvessel endothelial cells (BBMEC) verified that a close structural analog of Ziconotide (SNX-194) passed through this blood-brain barrier (BBB) model as expected for peptides of similar physical properties (permeability coefficient of 6.5 x 10(-4) cm/g). Passage from blood to brain was also verified by in situ perfusion through the carotid artery. A statistically greater amount of radioactivity was found to cross the BBB after perfusion of radioiodinated Ziconotide compared to [14C]inulin. Capillary depletion experiments and HPLC analysis defined the brain location and stability. PMID- 10822105 TI - Characterization of tynorphin, a potent endogenous inhibitor of dipeptidyl peptidaseIII. AB - To find a more effective inhibitor than spinorphin (LVVYPWT), an endogenous factor derived from bovine spinal cord, we synthesized spinorphin analogues and assayed their inhibitory activity toward DPPIII among enkephalin-degrading enzymes. Tynorphin (VVYPW), an N-terminal and C-terminal truncated form of spinorphin, exhibited more potent inhibitory activity and an IC50 value of 0.086 +/- 0.05 microg/ml (n = 4), whereas structures smaller than four amino acid residues exhibited almost no or less activity, suggesting that a five amino acid structure containing a Tyr-Pro residue is essential for the inhibition. The inhibition of DPPIII by tynorphin was predominantly competitive and the Ki value was found to be 7. 50 +/- 1.19 x 10(-8) M on Lineweaver-Burk plotting. The inhibitory activity of tynorphin toward other enkephalin-degrading enzymes such as neutral endopeptidase, aminopeptidase, and angiotensin-converting enzyme was not as high as that toward DPPIII, suggesting that tynorphin is a specific inhibitor of DPPIII. In HPLC analysis, human serum cleaved tynorphin rapidly (38% of control at 2 h and background level at 4 h), but in the presence of leuhisitin, an aminopeptidase inhibitor, tynorphin was maintained at the original level for 24 h. These results indicated that tynorphin had a more effective structure for expression of inhibitory activity toward DPPIII. PMID- 10822106 TI - Subcellular ontogeny of brain pyroglutamyl peptidase I. AB - The present work studies pyroglutamyl-peptidase I activity in several subcellular fractions of the developing brain. In the synaptosomal fraction, soluble pGlu peptidase I activity is low until postnatal Day 9 (with a peak at postnatal Day 2) and the activity increases at postnatal Day 15. In later stages there are not significant changes of synaptosomal pGlu-peptidase I activity. However, in the cytosolic fraction the activity is high from ED22 until postnatal Day 9, and afterwards decreases. The changes in the particulate fractions are generally less drastic. PMID- 10822107 TI - Hypothalamic galanin is up-regulated during hyperphagia and increased body weight gain induced by disruption of signaling in the ventromedial nucleus. AB - Disruption of signaling in the ventromedial nucleus (VMN) by colchicine (COL) produces transient (4 days) hyperphagia and weight gain. Microinjection of galanin into various hypothalamic sites stimulates feeding, so we tested the hypothesis that galanin is up-regulated in COL-treated rats by analyzing galanin concentrations in micropunched hypothalamic sites. Galanin was increased in the paraventricular nucleus on Days 1 through 4 after COL-injection. Galanin was also elevated in three other hypothalamic sites, the dorsomedial nucleus, lateral hypothalamic area, and perifornical hypothalamus, on Days 2-4 and in the lateral preoptic area, on Day 1 only. In the median eminence-arcuate nucleus and amygdala an initial decrease on Day 1 was followed by a then progressive increase through Day 4. These increases occurred despite marked elevations in blood insulin and leptin, hormones known to suppress hypothalamic galanin. When COL- or saline treated rats were injected intracerebroventricularly with galanin, it stimulated feeding further in the hyperphagic COL-treated rats, but the relative response over basal consumption was similar in both COL-treated and control rats. These results in VMN disrupted rats suggest that neurochemical rearrangements, including increased availability of galanin, may contribute to the hyperphagia and increased weight gain; additionally, it seems that neurons in the VMN normally exert a restraint on galanin signaling. PMID- 10822108 TI - Structure activity relationships for bradykinin antagonists on the inhibition of cytokine release and the release of histamine. AB - Highly potent bradykinin antagonists were found to inhibit bradykinin-induced release of cytokines but to stimulate histamine release. Both actions show structural requirements completely different from those for bradykinin B1 and B2 receptors, indicating that the release of some cytokines from spleen mononuclear cells and of histamine from rat mast cells is not mediated by these receptors. Most potent bradykinin antagonists release histamine at lower concentrations than does bradykinin itself. Dimers of bradykinin antagonists are the most potent compounds for histamine release. In contrast to enhanced histamine release, potent inhibition of cytokine release enhances the applicability of these compounds as anti-inflammatory drugs. Many of the peptides designed for high B2 receptor antagonism were found to be compared by their concentrations far more potent for inhibition of cytokine release than for smooth muscle contraction. Thus, for some antagonists inhibition of cytokine release was detected at concentrations as low as 10(-15) M. The rational design of peptide and nonpeptide bradykinin antagonists for therapeutic use requires not only knowledge about the potency but also knowledge about the structure-activity relationships of such important side effects as cytokine and histamine release. PMID- 10822109 TI - Enhanced ganglionic responses to substance P in spontaneously hypertensive rats. AB - Intravenous injection of substance P (SP) increases blood pressure in normotensive rats by stimulating sympathetic ganglia. This study compared the effects of SP to increase renal nerve firing and blood pressure in normotensive and hypertensive rats treated with chlorisondamine. The increase in renal nerve firing was greatest in spontaneously hypertensive rats (SHR), intermediate in Wistar rats, and least in Wistar-Kyoto (WKY) rats. Blood pressure was increased more in SHR than in Wistar rats. Blood pressure was not increased in WKY rats. Responses to the ganglionic stimulant 1,1-dimethyl-4-phenylpiperazinium were the same in the three strains. These results suggest that there is a selective increase in the action of SP on sympathetic ganglia of SHR and that ganglion responsiveness to SP is correlated with its effect on blood pressure. PMID- 10822110 TI - Characterization of functional endothelin receptors in the porcine myometrium. AB - To characterize the endothelin (ET) receptor that mediates the contraction induced by ET-1 in the porcine myometrium, we carried out a contraction study, radioligand binding study and molecular study (reverse transcription polymerase chain reaction) for detection of ET receptor-coding genes (mRNA). ET-1 (1 nM-1 microM) caused a tetrodotoxin-insensitive contraction in both longitudinal and circular muscles, but the longitudinal muscle was more sensitive to ET-1 than was the circular muscle. On the other hand, ET-3 and sarafotoxin S6c were less effective to cause a contractile response. The contraction induced by ET-1 was markedly inhibited by BQ123 and FR139317, but BQ788 only slightly inhibited the response induced by ET-1. The radioligand binding study indicated the presence of a single class of 125I-ET-1 binding sites with the same Kd value in both muscle layers. However, Bmax in the longitudinal muscle (3252 fmol/mg protein) was significantly higher than that in the circular muscle (1883 fmol/mg protein). ET 1 and FR139317 inhibited the specific 125I-ET-1 binding completely, but ET-3, sarafotoxin S6c and BQ3020 only slightly inhibited the specific binding (inhibition, 10-20%), suggesting that ET(A) is the dominant ET receptor subtype in the porcine myometrium. The results of the molecular study indicated the expression of both ET(A) and ET(B) receptor-coding genes in the porcine myometrium. In conclusion, ET-1 causes contraction of the porcine myometrium through activation of the ET(A) receptor present on smooth muscle cells. There is a marked muscle layer-related difference (longitudinal muscle > circular muscle) in the ET-1-induced contraction and the ET(A) receptor concentration. PMID- 10822111 TI - Lovastatin is a potent inhibitor of cholecystokinin secretion in endocrine tumor cells in culture. AB - Lovastatin prevents isoprene synthesis thereby affecting the structural organization of proteins involved in protein transport and secretion. Lovastatin at 1 microM decreases CCK 8 secretion by over 50% in WE cells and in CCK 8 expressing AtT20 cells. At 10 microM CCK 8 secretion was inhibited by two thirds and at 100 microM, cytotoxic effects were observed in both cell types. Addition of mevalonate does not restore CCK secretion and stimulation of secretion by forskolin is also partially inhibited. Cellular content of CCK 8 and pro-CCK were not altered in either of these cell lines except at 100 microM lovastatin. Our results clearly demonstrate that lovastatin at 1 microM strongly inhibits CCK 8 secretion at multiple levels while having little or no effect on its synthesis. This effect on secretion may be partly responsible for the adverse gastrointestinal side effects of lovastatin in patients. PMID- 10822112 TI - Effects of CGRP on human osteoclast-like cell formation: a possible connection with the bone loss in neurological disorders? AB - Osteoclast-like cell (OCL-like) differentiation is increased in long term cultures of bone marrow taken from paralyzed areas of paraplegic patients. Among the neuropeptides recently described in bone, calcitonin gene-related peptide (CGRP) has been shown in animal studies to inhibit bone resorption in vivo and OCL-like differentiation in vitro: its deficiency could thus be a link between the neural lesion and increased OCL-like production in paraplegia and some other neurologic disorders. We therefore investigated in this study the effects of CGRP on human OCL-like formation and found that it indeed has an inhibitory effect mediated at least in part via cAMP. PMID- 10822114 TI - Neurotensin inhibits neuronal Na+,K+-ATPase activity through high affinity peptide receptor. AB - Neurotensin is a peptide present in mammalian CNS and peripheral tissues, which may play a major role in neurotransmission or neuromodulation, subserving diverse physiological functions. We studied the effect of added neurotensin on ATPase activities in synaptosomal membranes isolated from rat cerebral cortex. Neurotensin at 3 x 10(-8)-3 x 10(-6) M concentration decreased 20-44% Na+,K+ ATPase activity but failed to modify Mg2+-ATPase activity; lower neurotensin concentrations (3 x 10(-14)-3 x 10(-10) M) had no effect on enzyme activities. This inhibitory effect was abolished by neurotensin heating, by enzyme preincubation with neurotensin during periods exceeding 10 min, or by adding 1 x 10(-6) M SR 48692, a high affinity neurotensin receptor antagonist. Levocabastine, which blocks low affinity neurotensin receptor, failed to alter enzyme inhibition by the peptide. It is suggested that the sodium pump may be a target for neurotensin effects at neuronal level involving the participation of high affinity neurotensin receptor. PMID- 10822113 TI - Orexin-A in the human brain and tumor tissues of ganglioneuroblastoma and neuroblastoma. AB - Regional distribution of orexin-A-like immunoreactivity in the human brain and pituitary, and the presence of orexin-A-like immunoreactivity in the tumor tissues of pheochromocytomas, ganglioneuroblastomas and neuroblastomas were studied by radioimmunoassay. Expression of orexin mRNA was studied by reverse transcriptase polymerase chain reaction (PCR) method. Orexin-A-like immunoreactivity was detected in every region of human brain, but not in the pituitary. The highest concentration of orexin-A-like immunoreactivity in the human brain was found in hypothalamus (17.8 +/- 4.3 pmol/g wet weight, mean +/- SEM, n = 7), followed by thalamus, medulla oblongata, and pons. Orexin-A-like immunoreactivity was detected in the tumor tissues of ganglioneuroblastoma and neuroblastoma, but not in the tumor tissues of pheochromocytoma. Reverse phase high performance liquid chromatographic analyses of the orexin-A-like immunoreactivity in the human brain extracts and neuroblastoma extracts showed a single immunoreactive peak, which was eluted in an identical position to synthetic human orexin-A. Orexin mRNA was expressed in the hypothalamus and in the tumor tissues of ganglioneuroblastoma and neuroblastoma. These findings suggest that orexin-A is produced in the hypothalamus and transported to various brain regions via axons. In addition, this study has shown for the first time the production of orexin-A by ganglioneuroblastomas and neuroblastomas. PMID- 10822115 TI - Binding of a pure 125I-monoiodoleptin analog to mouse tissues: a developmental study. AB - The preparation of a pure 125I-labeled monoiododerivative of mouse leptin is described. This radiolabeled analog has been used to characterize and localize central and peripheral leptin binding sites (Ob-R) of the mouse at different stages of its development. The affinity values found in membrane homogenates of various mouse tissues are similar and range between 0.1 and 0.3 nM, indicating that all the Ob-R isoforms have a similar affinity. Leptin binding sites are highly expressed at the membrane level in lung, intestine, kidney, liver, and skin and to a lesser degree in stomach, heart, and spleen. Brain, thymus, and pancreas homogenates are devoid of any specific binding. The distribution of mouse Ob-R has also been explored by autoradiography and dipping techniques on whole mouse sections. In lung, leptin binding sites are located at the pulmonary parenchyma and at the bronchiolar epithelial level. Binding sites are expressed all along the digestive tract from the tongue to the rectum (esophagus, stomach, intestine, colon, and rectum). In muscular visceral structures (stomach, intestine, and bladder) the binding is mainly present in the lamina propria. During development, leptin receptors are early expressed in the liver, kidney, and bone. In the lung, the Ob-R level increased gradually from birth to adulthood where the expression is maximal. By contrast, leptin receptors located in the medulla of the kidney remain remarkably constant all along the development. A broad signal is present in cartilage and bone particularly in vertebrae, limb, and ribs. Interestingly, leptin receptors are barely detectable in the mouse brain except in the choroid plexus and leptomeninges, whereas in the rat brain leptin binding sites are located in the thalamus, the piriform cortex, the cerebellum (at the granular and molecular cell layer), and the pineal gland. PMID- 10822116 TI - Fungal allergens and peptide epitopes. AB - Fungal allergens represent a major cause of atopic disorders. Immunochemical and molecular characterization of fungal allergens has been hampered by the lack of pure proteins and to inherent variation among fungal proteins and in their poor yields. With the advent of molecular biology techniques, a number of allergens have been cloned, sequenced, and expressed from a variety of fungal species. The knowledge of the primary, secondary, and tertiary structures of these allergens, the immunodominant regions of these proteins, and their interaction with T and B cell epitopes, results in better understanding of the molecular mechanisms of allergy and may provide avenues of immunologic intervention to treat patients. The present review deals with the current understanding of fungal allergen epitopes. PMID- 10822117 TI - Statistical considerations in the intent-to-treat principle. AB - This paper describes some of the statistical considerations in the intent-to treat design and analysis of clinical trials. The pivotal property of a clinical trial is the assignment of treatments to patients at random. Randomization alone, however, is not sufficient to provide an unbiased comparison of therapies. An additional requirement is that the set of patients contributing to an analysis provides an unbiased assessment of treatment effects, or that any missing data are ignorable. A sufficient condition to provide an unbiased comparison is to obtain complete data on all randomized subjects. This can be achieved by an intent-to-treat design wherein all patients are followed until death or the end of the trial, or until the outcome event is reached in a time-to-event trial, irrespective of whether the patient is still receiving or complying with the assigned treatment. The properties of this strategy are contrasted with those of an efficacy subset analysis in which patients and observable patient data are excluded from the analysis on the basis of information obtained postrandomization. I describe the potential bias that can be introduced by such postrandomization exclusions and the pursuant effects on type I error probabilities. Especially in a large study, the inflation in type I error probability can be severe, 0.50 or higher, even when the null hypothesis is true. Standard statistical methods for the analysis of censored or incomplete observations all require the assumption of missing at random to some degree, and none of these methods adjust for the potential bias introduced by post hoc subset selection. Nor is such adjustment possible unless one posits a model that relates the missing observations to other observed information for each subject-models that are inherently untestable. Further, the subset selection bias is confounded with the subset-specific treatment effect, and the two components are not identifiable without additional untestable assumptions. Methods for sensitivity analysis to assess the impact of bias in the efficacy subset analysis are described. It is generally believed that the efficacy subset analysis has greater power than the intent-to-treat analysis. However, even when the efficacy subset analysis is assumed to be unbiased, or have a true type I error probability equal to the desired level alpha, situations are described where the intent-to-treat analysis in fact has greater power than the efficacy subset analysis. The intent to-treat design, wherein all possible patients continue to be followed, is especially powerful when an effective treatment arrests progression of disease during its administration. Thus, a patient benefits long after the patient becomes noncompliant or the treatment is terminated. In such cases, a landmark analysis using the observations from the last patient evaluation is likely to prove more powerful than life-table or longitudinal analyses. Examples are described. PMID- 10822118 TI - Computations for group sequential boundaries using the Lan-DeMets spending function method. AB - We describe an interactive Fortran program which performs computations related to the design and analysis of group sequential clinical trials using Lan-DeMets spending functions. Many clinical trials include interim analyses of accumulating data and rely on group sequential methods to avoid consequent inflation of the type I error rate. The computations are appropriate for interim test statistics whose distribution or limiting distribution is multivariate normal with independent increments. Recent theoretical results indicate that virtually any design likely to be used in a clinical trial will fall into this category. Interim analyses need not be equally spaced, and their number need not be specified in advance. In addition to determining sequential boundaries using an alpha spending function, the program can perform power computations, compute probabilities associated with a given set of boundaries, and generate confidence intervals. PMID- 10822119 TI - Adjusting survival analysis for the presence of unadjudicated study events. AB - In the Coumadin Aspirin Reinfarction Study (CARS), the primary (efficacy) endpoint was time to the first occurrence of reinfarction, ischemic stroke, or cardiovascular death. Study events likely to be a component of the primary endpoint were reviewed by a blinded Events Classification Committee (ECC). To accommodate the inherent delay imposed by the adjudication process, we used the results of prior adjudication to predict classifications for selected unadjudicated study events in order to conduct "up-to-date" interim analyses for the Data and Safety Monitoring Committee (DSMC). Rates at which previously adjudicated study events were confirmed as primary event components were used to adjust the Kaplan-Meier estimates of survival and the log-rank statistics. Multiple events for a given subject were weighted according to the probability that prior events would be confirmed. Once all study events were adjudicated, final analyses confirmed the results of the adjusted interim analysis. For monitoring clinical trials with adjudicated endpoint events, models based on prior adjudication histories can be reliably used to predict event classifications for unadjudicated events and support accurate, contemporary interim analyses. PMID- 10822120 TI - A randomized trial assessing the impact of written information on outpatients' knowledge about and attitude toward randomized clinical trials. The INFO trial group. AB - To improve the patient education process in clinical research, three information materials describing general aspects of design and conduct of randomized clinical trials were developed. The materials varied in length, reading ability level, and reader appeal. Their influence on knowledge about and attitude toward randomized clinical trials was assessed in a randomized, parallel group, evaluator-blinded trial among 415 outpatients. The patients were randomized to the following groups: control (no intervention), leaflet, brochure, or booklet. Knowledge was assessed by a 17-item multiple-choice questionnaire and attitude was assessed by a 32-item Likert questionnaire at entry and 2 weeks after the intervention. The interventions and the questionnaires were pilot tested and power calculations were performed. At entry, the mean knowledge score was 7.9 points. At follow-up, the knowledge scores increased by 0.5 for the control, 1.0 for the leaflet, 1.6 for the brochure, and 1.4 for the booklet. The brochure and the booklet improved the knowledge score significantly compared with the control. The general attitude was positive at entry (mean 71.5 points). Only the booklet significantly increased the total attitude score (4.8 points) and the randomized clinical trials attitude subscale score (1.8 points). In conclusion, written information significantly improved outpatients' knowledge about and attitude toward randomized clinical trials. Detailed rather than brief information was more effective. Control Clin Trials 2000;21:223-240 PMID- 10822121 TI - Ruminations on the intent-to-treat principle. PMID- 10822122 TI - Design of the multicenter Australian study of epidural anesthesia and analgesia in major surgery: the MASTER trial. AB - The Multicenter Australian Study of Epidural Anesthesia and Analgesia in Major Surgery (The MASTER Trial) was designed to evaluate the possible benefit of epidural block in improving outcome in high-risk patients. The trial began in 1995 and is scheduled to reach the planned sample size of 900 during 2001. This paper describes the trial design and presents data comparing 455 patients randomized in 21 institutions in Australia, Hong Kong, and Malaysia, with 237 patients from the same hospitals who were eligible but not randomized. Nine categories of high-risk patients were defined as entry criteria for the trial. Protocols for ethical review, informed consent, randomization, clinical anesthesia and analgesia, and perioperative management were determined following extensive consultation with anesthesiologists throughout Australia. Clinical and research information was collected in participating hospitals by research staff who may not have been blind to allocation. Decisions about the presence or absence of endpoints were made primarily by a computer algorithm, supplemented by blinded clinical experts. Without unblinding the trial, comparison of eligibility criteria and incidence of endpoints between randomized and nonrandomized patients showed only small differences. We conclude that there is no strong evidence of important demographic or clinical differences between randomized and nonrandomized patients eligible for the MASTER Trial. Thus, the trial results are likely to be broadly generalizable. PMID- 10822123 TI - The estrogen replacement and atherosclerosis (ERA) study: study design and baseline characteristics of the cohort. AB - The Estrogen Replacement and Atherosclerosis (ERA) trial is a three-arm, randomized, placebo-controlled, double-blind trial to evaluate the effects of estrogen replacement therapy (0.625 mg/day oral conjugated estrogen) with or without continuous low-dose progestin (2.5 mg oral medroxyprogesterone acetate/day) versus placebo on progression of atherosclerosis. A total of 309 postmenopausal women at five sites underwent baseline coronary angiography and were randomized. Participants will have repeat coronary angiography after an average of 3.25 years of treatment. The primary outcome of interest will be change in minimum diameter of the major epicardial segments, as assessed by quantitative coronary angiography. The primary aim is to test the hypothesis that either form of hormone therapy will slow the progression or induce regression of coronary atherosclerosis compared to placebo. The secondary aims are to assess the effects of the two treatments versus placebo on endothelial function (measured using flow-mediated vasodilator responses), on several presumed mediators of estrogen's effect on atherosclerosis (i.e., plasma lipids and lipoproteins, blood pressure, glucose metabolism, hemostatic factors, and antioxidant activity), on other factors that influence the development of coronary heart disease (i.e., diet, smoking status, exercise, weight, and health related quality of life issues), and on clinical cardiovascular events. The ERA trial is the first angiographic endpoint clinical trial to examine the effects of postmenopausal hormone replacement on coronary atherosclerosis in women. It will provide an unparalleled opportunity to determine if either regimen of hormone therapy is effective in slowing the progress of angiographically defined coronary atherosclerosis. This study will complement other estrogen replacement trials, such as the PEPI, HERS, and Women's Health Initiative studies, to provide a more comprehensive examination of the effects of estrogen replacement on cardiovascular risk factors, anatomic and functional manifestations of atherosclerosis, and risk for coronary heart disease in postmenopausal women. Control Clin Trials 2000;21:257-285 PMID- 10822124 TI - The EARLY ALLIANCE prevention trial: a dual design to test reduction of risk for conduct problems, substance abuse, and school failure in childhood. AB - This paper describes a preventive intervention trial called EARLY ALLIANCE which is aimed at reducing risk for three adverse outcomes in childhood and adolescence: conduct problems, substance abuse, and school failure. The structure of the prevention trial is unique because two linked designs are being implemented concurrently. The primary design focuses on children at elevated risk for adverse outcomes, and compares a targeted, multicontextual preventive intervention with family, classroom, peer relational, and academic components to a universal, schoolwide preventive intervention that emphasizes peaceful conflict management and serves as a "usual care" control condition. The secondary design focuses on children at lower risk for adverse outcomes and compares a universally administered classroom program to the control condition. The paper describes the theoretical foundation for EARLY ALLIANCE, the goals of the prevention trial, the rationale for design choices, and the methods employed. PMID- 10822125 TI - The effect of 6-methylthiohexyl isothiocyanate isolated from Wasabia japonica (wasabi) on 4-(methylnitrosamino)-1-(3-pyridyl)-1-buatnone-induced lung tumorigenesis in mice. AB - The present study was undertaken to estimate the effect of 6-methylthiohexyl isothiocyanate (6MHITC) isolated from Wasabia japonica (wasabi) pretreatment on 4 (methylnitrosamino)-1-(3-pyridyl)-1-butanone(NNK)-induced lung tumorigenesis in mice. Pretreatment with 6MHITC for 4 consecutive days at a daily dose of 5 micromol significantly inhibited NNK-induced O(6)-methylguanine formation in lungs at 4 h after the injection. In conjugation with this inhibitory effect, 6MHITC suppressed the increase in proliferating nuclear cell antigen level as well as ornithine decarboxylase activity at a promotion stage of NNK-induced lung tumorigenesis. Finally, this treatment of 6MHITC suppressed the NNK-induced lung tumorigenesis in mice. These results suggest that 6MHITC inhibits the development of lung tumors in mice treated with NNK, due to the suppression of initiation stage. PMID- 10822126 TI - Thymosinalpha1 is chemopreventive for lung adenoma formation in A/J mice. AB - The effects of thymosin (THN) alpha1 were investigated using the urethane injection carcinogenesis A/J mouse model. Lung adenomas were observed 2.5, 3, and 4 months after urethane injection (400 mg/kg i.p.) into female A/J mice. Daily administration of THNalpha1 (0.4 mg/kg, s.c.) reduced lung adenoma multiplicity significantly, by approximately 45, 40, and 17%, respectively, 2.5, 3, and 4 months after urethane injection. Animals treated with THNalpha1 had a significantly greater white cell density than control A/J mice. Endogenous THNalpha1-like peptides were detected in the mouse lung. By radioimmunoassay and by Western blot, prothymosin alpha was detected in the mouse lung. By immunocytochemistry, THNalpha1-like peptides were detected in all lung compartments including the bronchus, adenoma, bronchioles, and alveoli. These results indicate that exogenous THNalpha1 prevents lung carcinogenesis in A/J mice. PMID- 10822127 TI - Effects of angiogenesis inhibitor TNP-470 on tamoxifen-stimulated MCF-7 breast tumors in nude mice. AB - The antitumor effect of TNP-470, antiangiogenic drug, was analyzed in the tamoxifen-stimulated MT-2 tumors inoculated in the athymic nude mice. TNP-470 was injected subcutaneously at a dose of 30 mg/kg body weight twice a week to mice which were randomized into three treatments: control (sham treatment), tamoxifen alone, and tamoxifen plus TNP-470. TNP-470 inhibited the growth of the tamoxifen stimulated MT2 tumors without any major side effects or significant weight loss compared with tamoxifen-treated mice alone. The mean tumor area of the mice treated with tamoxifen plus TNP-470 was reduced 50% to those treated with tamoxifen alone. TNP-470 was shown to inhibit tumor neovascularization and to increase incidence of apoptosis in tumor cells. TNP-470 did not affect tamoxifen metabolism of the mice. In conclusion, TNP-470 could be evaluated clinically in patients with tamoxifen failure. PMID- 10822128 TI - Involvement of SAPK/JNK pathway in X-ray-induced rapid cell death of human T-cell leukemia cell line MOLT-4. AB - We found that SAPK/JNK was phosphorylated during X-ray-induced rapid cell death of MOLT-4 cells and that acid Sphingomyelinase inhibitor D609 suppressed the rapid cell death as well as phosphorylation of SAPK/JNK. Also C2-ceramide caused phosphorylation of SAPK/JNK, followed by rapid cell death. Further we isolated X ray-resistant radiation-hybrid clones from MOLT-4 and 50 Gy irradiated mouse FM3A cells by repeated selections with 3 Gy irradiation. One of them named Rh-1a was found resistant to X-ray- as well as C2-ceramide-induced rapid cell death. Rh-1a cells had mouse DNA but no increase in either mouse or human Bcl-2 determined by Western blotting. Accumulation of p53 after X-irradiation was similarly observed in both parental MOLT-4 and Rh-1a cells. However, contrasting to prolonged and prominent phosphorylated status of SAPK/JNK in MOLT-4 cells, Rh-1a cells exhibited short transient increase and FM3A cells showed no increase of phosphorylated status SAPK/JNK after X-irradiation. Therefore, SAPK/JNK activation is considered important in X-ray-induced rapid cell death or apoptosis of MOLT-4 cells. PMID- 10822129 TI - Expression of thyroid hormone receptors is disturbed in human renal clear cell carcinoma. AB - Human renal clear cell carcinoma (RCCC) accounts for up to 2% of human cancers. To find out if thyroid hormone (T3) and its receptors (TRs) play a role in tumorigenesis of RCCC, the expression of TRs was evaluated on mRNA and protein level. It was found that TRalpha (both alpha1 and alpha2) mRNA amount was significantly decreased in tumors while compared with healthy kidney tissue, and this decrease was deepest in G1 (well differentiated) RCCCs. In contrast, TRalpha1 protein was 1.6x overexpressed in tumors. TRbeta1 mRNA amount was overexpressed in 30% and significantly decreased in 70% of examined tumors. On the protein level, TRbeta1 amount was 1.7x lower in tumors than in healthy controls. PMID- 10822130 TI - Establishment of a new human pancreatic cancer cell line, NOR-P1, with high angiogenic activity and metastatic potential. AB - We present here a new cell line, NOR-P1, established from a metastatic subcutaneous tumor of a patient with pancreatic cancer. The cells show rapid growth in culture with a doubling time of 16 h and high migration activity. Genetic and molecular analyses revealed high telomerase activity and a mutation in the K-ras oncogene. Of particular interest, the cells express markedly elevated mRNA levels of angiogenic factors, vascular endothelial growth factor and platelet-derived growth factor, as well as other tumor growth-related factors. Subcutaneous transplantation of the NOR-P1 cells into nude mice formed solid, hemorrhagic tumors which were histologically diagnosed as adenocarcinoma with dense blood vessels and severe extravasation of blood. Furthermore, when NOR P1 cell suspension was injected directly into the pancreas of nude mice, the cells grew rapidly to form intra-pancreatic tumors associated with liver metastases and peritoneal dissemination that resulted in cachexia and subsequent death. These properties suggest that NOR-P1 is an aggressive pancreatic cancer cell line with a high metastatic potential and may serve as a useful experimental model for studying tumor angiogenesis and metastasis of pancreatic cancer. PMID- 10822131 TI - Conventional tumor markers are prognostic indicators in patients with head and neck squamous cell carcinoma. AB - We tested for squamous cell carcinoma-related antigen (SCC), carcinoembryonic antigen (CEA), ferritin, immunosuppressive acid protein (IAP) and sialic acid in the serum from 247 patients with head and neck squamous cell carcinoma prior to therapy. Significant correlations were found between IAP and tumor size, lymph node metastasis, and clinical stage (P<0.0001, P<0.001, and P<0.0001). Also, sialic acid and SCC were also correlated with tumor size, lymph node metastasis, and clinical stage. Moreover IAP, sialic acid and SCC were strongly associated with survival rate (P<0.0001, P = 0.0230 and P = 0.0159). A multivariate Cox proportional hazard model demonstrated that being positive for IAP was an independent predictor for patients with H&NSCC (P = 0.0115). The results indicate that IAP, sialic acid and SCC are useful as prognostic factors. PMID- 10822132 TI - Predictive value of serum immunosuppressive acidic protein for lung metastasis after amputation of murine osteosarcoma of the lower limb. AB - A spontaneously occurring murine osteosarcoma cell line (POS-1) was inoculated into the footpads of 88 mice. In 59 mice, the tumor-bearing leg was amputated at 1, 3, 5, or 7 weeks after inoculation and the mice sacrificed at 9 weeks. Lung metastasis was observed in 82.6-100% of the mice undergoing leg amputation at 5 or 7 weeks after inoculation. There were no lung metastasis in the mice amputated at 1 or 3 weeks. The serum immunosuppressive acidic protein (IAP) concentration showed a significant increase at 5 or 7 weeks after inoculation (P<0.005 and P<0.0005, respectively). When amputation was done 5 weeks after inoculation, the serum IAP concentration increased further to 5.9 times the pre-amputation level 1 week after the amputation. The IAP concentration increased only 1.3-fold in the control group without inoculation. Thus, a high serum IAP concentration before amputation and a further increase after amputation were indicators of lung metastasis in this mouse model of osteosarcoma. PMID- 10822133 TI - Expression of membrane-type 1 matrix metalloproteinase (MT1-MMP) on prostate cancer cell lines. AB - Membrane-type metalloproteinase-1 (MT1-MMP) is a transmembrane metalloproteinase, which activates proMMP-2 and expressed on the cell surface in many invasive cancer cells. We investigated the expression of MT1-MMP in prostate cancer cell lines. MT1-MMP protein and mRNA were expressed in PC-3, DU-145 and TSU-pr1 cells (androgen-independent prostate cancer cell lines), but in LNCaP cells (androgen dependent prostate cancer cell line). MT1-MMP protein was negative and mRNA was low to detect by RT-PCR. Cell lysate of PC-3 cleaved proMMP-2 to the active form. In addition, both hepatocyte growth factor (HGF) and gastrin-releasing peptide (GRP) increased Matrigel invasion and induced the expression of MT1-MMP protein in DU-145 prostate cancer cells. These results suggest that MT1-MMP is indeed the tumor-specific activator of proMMP-2 in androgen-independent prostate cancer cells and plays an important role in the invasive properties of prostate cancer cells. PMID- 10822134 TI - Gains in chromosomes 7, 8q, 15q and 17q are characteristic changes in malignant but not in benign peripheral nerve sheath tumors from patients with Recklinghausen's disease. AB - In order to investigate typical genomic alterations in patients with Recklinghausen's disease (NF1) we studied one from each of the six patients with NF1 several benign and/or malignant tumors. By means of comparative genomic hybridization (CGH) gained results from six benign neurofibromas and 14 malignant peripheral nerve sheath tumors (MPNSTs) were compared with four benign peripheral nerve sheath tumors (BPNSTs) from patients without NF1. In all 14 MPNSTs DNA sequence copy number changes were detected with a mean value of 13.5 imbalances per sample. The most frequent gains were in 8q, 17q (12 tumors each), 7p, 15q (ten tumors each), and 7q (nine tumors). We found ten high-level amplifications in nine of the 14 samples. In two cases, the high-level amplification involved 7p14-pter and 17q24-qter as well. The most frequent loss was in 17p (seven tumors). The benign neurofibromas from NF1-patients and the sporadic BPNSTs revealed only partially DNA sequence copy number changes without any distinct pattern. The gains of #7, 8q, 15q, and 17q were found exclusively in MPNSTs but not in neurofibromas and are supposed to be associated with malignant tumor progression. In comparison of the results of the 14 MPNSTs from NF1-patients with the results of previously published 20 sporadic MPNSTs, we found that the gain of 8q occurs most frequently in both tumor groups. Of course additionally in the sporadic MPNSTs there were more frequent gains of 5p, #6, and statistically significant gains of 20q. On the other hand in the MPNSTs from NF1-patients the most frequent gains were found in #7, and statistically significant in 15q, and 17q. PMID- 10822135 TI - Caspase involved synergistic cytotoxicity of bcl-2 antisense oligonucleotides and adriamycin on transitional cell cancer cells. AB - We have previously shown that Bcl-2 expression was negative prognostic factor in transitional cell cancer (TCC), and that TCC cell lines expressing high levels of Bcl-2 are resistant to Adriamycin triggered apoptosis. Here we examined antisense oligonucleotide-mediated downregulation of Bcl-2 expression and its effect on sensitivity to Adriamycin (ADM) treatment in T24 cells. Treatment of T24 cells with 20 microM of bcl-2 antisense phosphorothioate oligodeoxynucleotide (PODN) reduced the Bcl-2 protein level. Combined administration with Adriamycin resulted in synergistic cytotoxicity, accompanied with a 2.4-fold increase in DEVD specific caspase activity. The finding provides evidence that Bcl-2 expression may be critical for maintaining the drug resistance of TCC. bcl-2 antisense PODN might be useful means for overcoming drug resistance in highly malignant TCC. PMID- 10822136 TI - Differential inhibition of renal cancer cell invasion mediated by fibronectin, collagen IV and laminin. AB - Invasion of tumor cells into the extracellular matrix is an essential step in the formation of metastases in renal cancer. Cell adhesion molecules such as beta(1) integrins, which bind to the RGD sequence (arginine-glycine-asparagine) and CD44 are involved in this process. We examined the invasion of a renal carcinoma cell line (CCF-RC1) into the extracellular matrix compounds fibronectin, collagen IV and laminin and the effect of TGFbeta and IFNgamma on this process. The inhibitory effect of an antibody against the beta(1)-subunit of integrins (CD29), as well as a pentapeptide including the RGD sequence, was also evaluated. A micro chemotaxis chamber, including a polycarbonate membrane with a pore diameter of 8 microm, was used for quantification of cell migration. The addition of the extracellular matrix compounds fibronectin, laminin and collagen IV resulted in a 5-10-fold increase in invasion. This increased invasion depends strongly on the presence of beta(1)-integrins, shown by the use of an antibody against CD29 or a RGD including peptide which inhibit the cell migration by approximately 88%. CD44 is less involved in collagen IV dependent migration and almost no influence of CD44 was observed on a fibronectin and laminin dependent migration. TNFalpha and IFNgamma did not significantly influence the expression of CD29 or CD44, and no alteration in tumor cell migration was observed. These results show that the invasion of renal cancer cells is differentially regulated by compounds of the extracellular matrix, whereby fibronectin seems to be the most critical factor. The molecular interactions in this process are strongly dependent on beta(1) integrins and the corresponding amino acid sequence RGD. PMID- 10822138 TI - Brain oestradiol and testosterone levels in Alzheimer's disease. AB - Epidemiological studies indicate that oestrogen improves memory and may delay the onset of Alzheimer's disease (AD) in postmenopausal women. Furthermore, evidence from experimental studies suggests beneficial effects of oestrogen on several pathogenic mechanisms implicated in AD. We have therefore measured the levels of oestradiol and testosterone in control and AD brains. The results show that in control brain, oestradiol levels are 3.5 fold higher in females than males, though testosterone levels are equivalent. In AD, oestradiol levels were not significantly increased compared to those in control brain, while testosterone levels were unaffected in AD. The results do not support the hypothesis that a lack of oestrogen is a contributory factor in AD. PMID- 10822139 TI - Isovolumetric regulation of rat glial cells during development and correction of hypo-osmolality. AB - Rat C6 glioma cells undergo regulatory volume decrease (RVD) following sudden exposure to hypo-osmolality, but little or no regulatory volume increase (RVI) is observed when cells cultured in hypo-osmotic media are suddenly returned to isoosmolality. Because C6 glioma cells would rarely be exposed to sudden large changes in osmolality in vivo, we examined the ability of these cells to maintain their volume, termed 'isovolumetric regulation', when exposed to gradual changes in osmolality. When osmolality was gradually reduced by reduction of NaCl concentration from 300 to 250 mOsmol/kg at a rate of 0.4 mOsmol/kg/min or less cells were able to maintain their volume, while at higher rates, the cells swelled. Cells which were cultured in hypo-osmotic (200 mOsmol/kg) media for 3 days exhibited isovolumetric regulation at rates of osmolality increase of 0.5 mOsmol/kg/min or less over the range of 200-250 mOsmol/kg. We conclude that rat C6 glioma cells can sensitively regulate their volume over the osmolality range of pathophysiologic interest at rates of osmolality change which are faster than those generally seen in clinical conditions. PMID- 10822140 TI - Relative electroencephalographic desynchronization and synchronization in humans to emotional film content: an analysis of the 4-6, 6-8, 8-10 and 10-12 Hz frequency bands. AB - The reactivity of different narrow electroencephalographic (EEG) frequencies (4 6, 6-8, 8-10 and 10-12 Hz) to three types of emotionally laden film clips (aggressive, sad, neutral) were examined. We observed that different EEG frequency bands responded differently to the three types of film content. In the 4-6 Hz frequency band, the viewing of aggressive film content elicited greater relative synchronization as compared the responses elicited by the viewing of sad and neutral film content. The 6-8 Hz and 8-10 Hz frequency bands exhibited reactivity to the chronological succession of film viewing whereas the responses of the 10-12 Hz frequency band evolved within minutes during film viewing. Our results propose dissociations between the responses of different frequencies within the EEG to different emotion-related stimuli. Narrow frequency band EEG analysis offers an adequate tool for studying cortical activation patterns during emotion-related information processing. PMID- 10822141 TI - Prenatal and postnatal exposure to ethanol induces changes in the shape of the dendritic spines from hippocampal CA1 pyramidal neurons of the rat. AB - 20%-ethanol was provided to adult female rats since a pregestational stage until weaning of the pups, and percentage proportion of thin, mushroom-shaped, stubby, or wide spines from the apical dendrite of hippocampal CA1 pyramidal cells, was counted at 15, 21, 40, and 90 days of age. By-kind-of-spine analysis revealed higher fluctuation of experimental spines, and less percentage of thin spines was observed in the ethanol-intoxicated rats concomitantly with a higher proportion of stubby or wide spines; through development. Because thin spines may propagate the synaptic potentials more efficiently than stubby or wide spines, this findings suggest that the electrical excitability and thereafter the firing pattern of those cells may be altered, due to the toxic effects of chronic ethanol ingestion. PMID- 10822142 TI - An anterograde degeneration study of the distribution of regenerating rat myelinated fibers in the silicone chamber model. AB - Specificity of reinnervation after a peripheral nerve lesion has given rise to considerable controversy. As a contribution to solving this issue we have evaluated the specificity of reinnervation of the peroneal nerve after a complete transection of the sciatic nerve repaired with an 8 mm silicone tube, leaving a 4 mm gap between the nerve stumps. Our findings reveal unspecificity of reinnervation of the distal peroneal branch. This lack of specificity is shown by a random distribution of fibers originating from both proximal branches at the level of the tube and at distal peroneal and tibial branches, argue against specificity of regeneration in this model. PMID- 10822143 TI - Spatial learning affects immature granule cell survival in adult rat dentate gyrus. AB - Neurogenesis occurs throughout life in mammalian dentate gyrus. The effect of learning on newborn cell survival was studied in rat. Rats were trained on a hippocampus-dependent spatial learning task by using Morris water maze. Neurogenesis was evaluated by 5-bromo-2'deoxyuridine administered before learning. Several newborn cells expressed the immature neuron marker TOAD-64. The main findings were as follows: (1) the survival of newborn cells was enhanced by learning at early stage of differentiation; (2) the newborn cells saved by learning were mainly located in the rostral part of external blade of granule cell layer and (3) there was a correlation between the actual individual learning and newborn cell survival. PMID- 10822144 TI - The taurine uptake inhibitor guanidinoethyl sulphonate is an agonist at gamma aminobutyric acid(A) receptors in cultured murine cerebellar granule cells. AB - In patch clamp experiments the beta-amino acid uptake inhibitor guanidinoethyl sulphonate (GES) activated currents in intact cultured murine cerebellar granule neurones. These responses could be attenuated by the gamma-aminobutyric acid(A) (GABA(A)) receptor antagonists bicuculline and picrotoxin. With intracellular chloride concentrations of either 20 or 130 mM, GES-induced current responses reversed polarity near the chloride equilibrium potential. When fast applications of agonist were made to excised granule cell macropatches GES responses were dose dependent and exhibited significant outward rectification. Like taurine (but unlike GABA and beta-alanine) responses, macroscopic desensitisation of GES induced currents was slow. Our data indicate that care should be exercised when using GES as a taurine uptake inhibitor in systems that also contain GABA(A) receptors. PMID- 10822145 TI - Identification of the primary auditory field in archival human brain tissue via immunocytochemistry of parvalbumin. AB - Using parvalbumin (PV)-immunocytochemistry, we identified the human primary auditory field (PAF) for the first time. The first gyrus of the right Heschl was identified, and a serial perpendicular cut to the gyrus was made. PV immunoreactive neuropils delineated four zones, as have been described in monkeys. Unlike with conventional staining, zone 1, characterized by intense immunostaining, corresponded to the koniocortex (PAF), while zone 2, characterized by moderate immunostaining around zone 1, to the parakoniocortex. PV-immunocytochemistry was a useful way to identify PAF in formaline-fixed archival human brain tissue. PMID- 10822146 TI - Adenosine triphosphate-induced heterologous desensitization of endothelin-1- and glutamate-evoked calcium increases in cultured rat cortical astrocytes. AB - In rat cortical astrocytes, we investigated the occurrence of cross-talks between purinoceptor and endothelin (ET) receptor, or glutamate receptor. The treatments of adenosine triphosphate (ATP), ET-1, and glutamate induced the increase of intracellular calcium level in the astrocytes. In repetitive additions of ATP to astrocytes, the second application of ATP exhibited comparable amplitude of calcium response, but the stimulation with ATP completely blocked subsequent ET-1 or glutamate-evoked calcium responses showing complete heterologous desensitization. In contrast, ET-1 and glutamate failed to desensitize the response elicited by ATP. Preincubation with sphingosine, a protein kinase C (PKC) inhibitor, reversed the ATP-induced desensitization of ET-1- and glutamate evoked calcium responses. Taken together, these results demonstrate the resistance of purinoceptor to homologous desensitization, and unidirectional desensitization between ATP and other receptors such as ET and glutamate receptors, suggesting a dominant role of purinoceptor in modulating calcium signal of astrocytes. PMID- 10822147 TI - Potentiation of morphine and clomipramine analgesia by cholecystokinin -B antagonist CI-988 in diabetic rats. AB - The aim of this study was to determine the influence of an intrathecally injected cholecystokinin-B (CCK-B) receptor antagonist, CI-988, on the analgesic effect of morphine and clomipramine in diabetic rats. Administered alone, morphine (0.1 mg/kg, i.v.) and clomipramine (3 mg/kg, i.v.) have respectively no effect and only a slight effect on vocalization thresholds to paw pressure in diabetic rats, but, when coadministered with CI-988 (0.1 microg/rat, i.t.), an appreciable antinociceptive effect was observed. This suggests that a spinal blockade of cholecystokininergic system increases the analgesia induced by morphine or clomipramine. A CCK-B receptor antagonist could thus be used to lower dosages of morphine or antidepressant drugs in the management of neuropathic pain in humans, and thereby reduce their side effects. PMID- 10822148 TI - Increased immunoreactive labeling of the spinal N-methyl-D-aspartate R1 receptors after dorsal root ganglionectomy in the rats. AB - The N-methyl-D-aspartate (NMDA) receptor plays an important role in the development of the autotomy after dorsal root ganglionectomy (DRGn). In this study, we further investigated the expression of the NMDAR1 in the spinal cord of the rats after right DRGn by immunohistochemical analyses. Computerized densitometric analysis of the NMDAR1 immunoreactivity was done and the integrated optical density (IOD) of the superficial laminae of the dorsal horn of the spinal cord was measured. The immunoreactive labeling of the NMDAR1 was increased in the cervical spinal cord ipsilateral to the DRGn from day 5 to 14 after DRGn. The ratio of the right/left IOD of the rats receiving DRGn was significantly higher than the rats in the sham-operated group and the control group (P<0.05). The expression of the NMDAR1 increased gradually to reach the peak at day 7 after DRGn (mean right/left IOD ratio=1.52), then decreased thereafter. The increased expression of the NMDAR1 at day 7 was suppressed by MK-801 (NMDA receptor antagonist) administered immediately after DRGn, but not by normal saline or 1,2,3,4-tetrahydro-6-nitro-2, 3-dioxo-benzo[f] quinoxaline-7-sulfonamide (NBQX, non-NMDA receptor antagonist). The results indicated that the expression of the NMDAR1 in the superficial laminae of the dorsal horn of the spinal cord was increased after DRGn and the time course was compatible with the onset and development of the autotomy induced by DRGn. PMID- 10822150 TI - The dynamics of human isometric pointing movements under varying accuracy requirements. AB - The goal of the present study was to explore the relation between speed-accuracy trade-off phenomena and action kinematics in the case of pointing under isometric conditions. Increasing task difficulty resulted in a linear increase in movement time (as predicted by Fitts' law) and in systematic changes in the spatio temporal patterning of force production. The observed changes in motion topology were similar to those reported for isotonic tasks and adequately captured by a limit cycle model derived for the latter type of task. These results indicate that, for isometric force control as for isotonic position control, the reasons underlying the emergence of Fitts' law might be sought in dynamic trajectory formation processes. PMID- 10822149 TI - Effect of adenosine on adenosine triphosphate-sensitive potassium channel during hypoxia in rat hippocampal neurons. AB - Using the whole-cell patch clamp method, we explored the effect of adenosine on the K(ATP) current and its regulatory mechanisms in acutely dissociated rat hippocampal neurons. A chemical hypoxia model was made using 0.2 mmol/l 2,4dinitrophenol (2,4DNP). During hypoxia, the K(ATP) current was not raised significantly by adenosine alone, but was accelerated significantly by adenosine in combination with the selective A(2) receptor blocker 3, 7-dimethl-1 propargylxanth-ine. The selective A(1) receptor agonist N6-cyclopentyladenosine also accelerated the K(ATP) current. These results suggest that activation of the adenosine A(1) receptor can accelerate opening of the K(ATP) channel during hypoxia, and that the A(2) receptor may have an opposing effect to the A(1) receptor. PMID- 10822152 TI - Dorsal-ventral differentiation of short-term synaptic plasticity in rat CA1 hippocampal region. AB - Two forms of short-term synaptic plasticity (STP), paired-pulse facilitation (PPF) and frequency potentiation (FP) of CA1 field excitatory postsynaptic potentials (EPSP) to afferent stimulation were compared in slices taken from the dorsal and ventral parts of rat hippocampus. While dorsal slices showed significant PPF at all interpulse intervals (20-1400 ms, 80% at 40 ms), PPF in ventral slices was substantially weaker at intervals shorter than 100 ms (19% at 40 ms) and nil at longer intervals. While dorsal slices showed substantial FP at frequencies 1-40 Hz and frequency depression at 50-100 Hz, ventral slices showed only a much smaller potentiation at 1 Hz and substantial depression at 20-100 Hz. Decreasing [Ca(2+)](o) from 2 to 1 and 0.5 mM substantially reduced the baseline EPSPs in both groups of slices but its effect on PPF was greater in ventral slices. On the contrary when [Ca(2+)](o) was increased to 5 mM only dorsal slices showed an enhancement of baseline EPSP. It is concluded that ventral hippocampus CA1 area has a specific deficit in STP, which is related to the important presynaptic role of calcium and is consistent with a relatively higher transmitter release probability. PMID- 10822151 TI - Activation of pre- and postsynaptic protein kinase C during tetraethylammonium induced long-term potentiation in the CA1 field of the hippocampus. AB - Tetraethylammonium (TEA) induces a form of long-term potentiation (LTP) that is independent on N-methyl-D-aspartate (NMDA) receptor activation (LTP(K)). LTP(K) may be a suitable chemical model to study molecular mechanisms underlying LTP. We monitored the phosphorylation state of two identified neural-specific protein kinase C (PKC) substrates (the presynaptic protein GAP-43/B-50 and postsynaptic protein RC3) after different chemical depolarisations. TEA induced a long-lasting increase in synaptic efficacy in the CA1 field of the hippocampus and increased the phosphorylation of both GAP-43/B-50 and RC3 (51 and 56.1%, respectively). These effects were blocked by the voltage-dependent calcium channel antagonist nifedipine, but not by the NMDA receptor antagonist AP5. These data show that in LTP(K) the in situ phosphorylation of pre-and postsynaptic PKC substrates is increased, indicating that NMDA receptor-dependent and NMDA receptor-independent LTP share common Ca(2+)-dependent expression mechanisms, including activation of pre- and postsynaptic PKC. PMID- 10822153 TI - Role of the vestibular system in sudden shutdown of renal sympathetic nerve activity during microgravity in rats. AB - The purpose of this study was to examine the effect of microgravity (muG) on renal sympathetic nerve activity (RNA) in rats. Additionally, we estimated the participation of the vestibular system in the response of RNA to muG. Eight normal Sprague-Dawley (SD) rats and five chemically and bilaterally labyrinthectomied SD rats were used to measure RNA during free-drop examination (4.5-s duration of muG); arterial pressure (AP) and aortic flow velocity (AFV) were additionally monitored. Although AFV showed no particular change, AP tended to decrease during muG in the later phase. Prior to this AP fall-off, RNA was immediately and markedly attenuated by muG. This attenuation was transient and RNA returned to 1G level within the mu;muG condition. Interestingly, this phenomenon remained even in labyrinthectomied rats. In conclusion, cephalad shift of the body fluid by loading of muG may cause cardiopulmonary low-pressure receptor activation and consequent RNA attenuation, but the participation of the vestibulosympathetic reflex in this phenomenon is not obvious. PMID- 10822154 TI - Do brain oscillations of different frequencies indicate interaction between cortical areas in humans? AB - Oscillatory cortical activity in the beta frequency range (14-32 Hz) was investigated in a voluntary movement task using closely-spaced electroencephalogram recordings over the sensorimotor cortex. According to recent studies, short-lasting beta oscillations were observed within the first second after termination of a self-paced finger movement. In this study we show, moreover, that finger movement creates beta bursts not only overlying the hand representation area, but also, at a higher frequency, over neighboring cortical areas representing the foot. The results indicate that, related to the same event, distinct cortical areas can display increased synchronized activity at different frequencies, providing evidence for interaction between the corresponding neuronal networks. PMID- 10822155 TI - Degeneration process of Lewy bodies in the brains of patients with dementia with Lewy bodies using alpha-synuclein-immunohistochemistry. AB - We investigated the degeneration process of Lewy bodies (LB) in the brains of dementia with Lewy bodies, using alpha-synuclein-immunohistochemistry. Intracellular LB, LB-related neurites and some extracellular LB were positively immunostained with anti-alpha-synuclein antibodies. Concentric LB-bearing neurons had no microglial involvement, while degenerated neurons with ill-defined LB displayed intense microglial involvement. The late stage of extracellular LB were immunoelectron-microscopically composed of loose aggregates of filamentous components with lost alpha-synuclein-immunoreactivity and penetrated astroglial processes. These findings suggest that microglias are involved during the stages from degenerated LB-bearing neurons to extracellular LB, while astroglias are involved during the stage of extracellular LB. Some intracellular LB were positive for anti-C3d and -C4d antibodies, suggesting that the classical complement pathway is activated in degenerated LB-bearing neurons, inducing microglial activation and neuronal death. PMID- 10822156 TI - Nitric oxide: an inhibitor of NF-kappaB/Rel system in glial cells. AB - Nitric oxide (NO) has been reported to regulate NF-kappaB, one of the best characterized transcription factors playing important roles in many cellular responses to a large variety of stimuli. NO has been suggested to induce or inhibit the activation of NF-kappaB, its effect depending, among others, on the cell type considered. In this review, the inhibitory effect of NO on NF-kappaB (and subsequent suppression of NF-kappaB-dependent gene expression) in glial cells is reported. In particular, exogenous and endogenous NO has been observed to keep NF-kappaB suppressed, thus preventing the expression of NF-kappaB-induced genes, such as inducible NO synthase itself or HIV-1 long terminal repeat. Furthermore, the possible molecular mechanisms of NO-mediated NF-kappaB inhibition are discussed. More specifically, NO has been reported to suppress NF kappaB activation inducing and stabilizing the NF-kappaB inhibitor, IkappaB alpha. On the other hand, NO may inhibit NF-kappaB DNA binding through S nitrosylation of cysteine residue (i. e., Cys62) of the p50 subunit. As a whole, a novel concept that the balance of intracellular NO levels may control the induction of NF-kappaB in glial cells has been hypothesized. PMID- 10822157 TI - Medullary noradrenergic neurons projecting to the bed nucleus of the stria terminalis express mRNA for the NMDA-NR1 receptor. AB - The bed nucleus of the stria terminalis pars ventralis (vBNST) receives dense noradrenergic terminals and contains the highest concentration of noradrenaline (NA) in the brain. We used autoradiography following retrograde axonal transport of [(3)H]-NA to identify selectively whether noradrenergic neurons innervating the vBNST originate in the medulla oblongata and/or the locus coeruleus. In combination with this technique, non-isotopic in situ hybridization for the NMDA NR1 receptor subunit mRNA was used to examine, on the same brain sections, its expression in noradrenergic neurons that innervate the vBNST. The results showed that 60 +/- 6% and 35 +/- 7% of the total number of radiolabeled cells detected after injection of [(3)H]-NA in the vBNST were located in brainstems A1 and A2 noradrenergic cell groups, respectively. In addition, 18.5 +/- 4.2% of radiolabeled cells in A1 and 15.7 +/- 5% in A2 also expressed the mRNA for the NMDA-NR1 receptor subunit. In contrast, only 4 +/- 3% of the radiolabeled cells were present in the locus coeruleus, and none of these cells was positive to NMDA NR1 receptor subunit mRNA. The present results provide evidence that BNST noradrenergic fibers and terminals originate predominantly from A1 and A2 noradrenergic cell groups, and that a significant number of these noradrenergic neurons also express the mRNA for the NMDA-NR1 receptor subunit. The observation that brainstem noradrenergic neurons innervating the vBNST express NMDA receptor mRNA gives anatomical support to the regulation of NA release by NMDA presynaptic receptors. PMID- 10822158 TI - Induction and adaptation of Fos expression in the rat brain by two types of acute restraint stress. AB - The present study was designed to examine whether both induction and adaptation of brain Fos expression during acute stress depend on the intensity and duration of stressors. For this purpose, different durations of two types of acute stress, mild (restraint) and severe (immobilization) stress, were employed. Stress induced Fos expression was analyzed quantitatively by immunohistochemistry. Adaptation of Fos expression to the acute stressors was not apparent in the hypothalamic paraventricular nucleus (PVN) or locus coeruleus (LC) but was observed in the amygdala, hippocampus, and cerebral cortex. A higher level of Fos expression was seen in the PVN, LC, and amygdala, following severe stress than was seen following mild stress. In the hippocampus, the dentate gyrus showed reduced Fos expression in response to stressors, although both mild and severe acute stress increased Fos expression in other regions of the hippocampus. The cingulate cortex showed increased Fos expression during mild stress, whereas long duration severe stress reduced Fos expression. In the somatosensory cortex, both stressors increased Fos expression. These results indicate that the PVN and LC are relatively resistant to adaptation to acute stress compared to other brain regions. In addition, the PVN, LC, and amygdala may play important roles in the perception of the severity of stress. PMID- 10822159 TI - The effect of pyramidal stimulation upon tail muscle motoneurons in the decerebrate cat. AB - This study aimed to determine the effects of the corticospinal tract (CST) on the motoneurons innervating the tail muscles in cats. The stimulation of the pyramidal tract predominantly evoked excitatory postsynaptic potentials (EPSPs; 48/90 motoneurons: 53%). Single-pulse stimulation produced EPSPs in 18 of 48 motoneurons, but double shocks evoked postsynaptic potentials in most of the remaining cells (26/48). Monosynaptic excitatory connections between pyramidal tract fibers and tail motoneurons were confirmed in 4 motoneurons. Inhibitory postsynaptic potentials (IPSPs) were recorded from motoneurons innervating long tendinous tail muscles (7/90: 8%) and the shortest neuronal pathways of IPSPs were shown to be disynaptic pathways. Interactions between the CST and reflex pathways from low-threshold muscle and cutaneous afferents innervating the tail and hindlimbs were observed. PMID- 10822160 TI - Immunohistochemical estimation of brain choline acetyltransferase and somatostatin related to the impairment of avoidance learning induced by thiamine deficiency. AB - We have found that thiamine-deficient (TD) rats show significant impairment of avoidance learning on the 25th day after the start of TD diet, as measured by passive-avoidance task. Administration of physostigmine (0.1 mg/kg, i.p.) from the 14th day after the start of TD diet improved the impairment of avoidance learning to the pair-fed (PF) control level by the 25th day. However, the recovery effect of physostigmine did not occur on the 25th day when the treatment was begun on the 21st day. To ascertain the correlation between the cholinergic neuronal function in rat brain and the avoidance learning impairment induced by TD, the immunohistochemical distribution of brain choline acetyltransferase (ChAT) was determined by fluorescence intensity using two-dimensional microphotometry. The intensity of the ChAT fluorescence started to decrease in the cortex and hippocampus on the 14th day and showed a marked decrease in the cortex, hippocampus and thalamus on the 25th day of TD feeding in comparison with PF controls. The intensity of the somatostatin (SST) fluorescence was unchanged on the 14th day of TD feeding, but on the 25th day, SST was significantly decreased in comparison with PF controls. Furthermore, physostigmine treatment from 14th day after the start of TD diet reversed SST fluorescence intensity to the control level by the 25th day. These results suggest that the impairment of avoidance learning induced by TD may involve not only cholinergic but also somatostatinergic systems. PMID- 10822161 TI - Serotonergic and kynurenic pathways in rats exposed to foot shock. AB - Electric foot shock was applied to rats and levels of tryptophan and its metabolites were measured in the plasma, central nervous system and peripheral tissues. Metabolites of tryptophan are the results of the enhancement of serotonergic and kynurenine pathways. Plasma levels of tryptophan increased significantly immediately after the foot shock and returned to normal values within 24 h. Tryptophan levels also increased in all the brain areas immediately after stress application and returned to normal values within 24 h. Foot shock elevated the levels of kynurenine in the plasma, liver, kidney and every parts of the brain. 3-Hydroxykynurenine and kynurenic acid levels were increased in the brain. The present observations suggest that stress activates not only serotonergic pathway but also kynurenine pathway in the central nervous system and periphery. Some metabolites of kynurenine pathway, such as 3 hydroxykynurenine, are neurotoxic while other metabolite, such as kynurenic acid, may be neuroprotective. Increase in serotonin level in the hypothalamus and midbrain stabilises emotion and prevents mood disorders. Therefore, some brain dysfunction resulting from stress may be prevented by the metabolites of tryptophan. The balance of these functions may be important in the maintenance of nerve integrity under stress conditions. PMID- 10822162 TI - Phospholipid composition and levels are not altered in fibroblasts bearing presenilin-1 mutations. AB - Lipid alterations have been reported in brain regions affected by Alzheimer disease (AD). The mechanisms causing these changes are poorly understood because it is difficult to study dynamic, biochemical processes in post-mortem brain. Fibroblasts derived from AD patients offer an alternative model to study disease related alterations in lipid metabolism. Therefore, we measured the phospholipid levels and composition of fibroblasts from individuals bearing two different presenilin-1 mutations and compared these values to appropriate control fibroblasts. There were no differences between groups in phospholipid composition or in individual phospholipid levels, including the plasmalogens. Cholesterol levels and the cholesterol/phospholipid ratio were not different between presenilin-1 mutation bearing and control fibroblasts. Although these presenilin 1 mutation bearing fibroblasts have a number of biochemical changes related to AD, the absence of a change in phospholipid levels suggests that under these conditions, these cells are not useful in studying the mechanisms underlying the alterations in brain phospholipid levels associated with AD. However, these results do not preclude the possible use of other fibroblasts bearing AD-related mutations, e.g., APP mutations, to examine AD-related changes in brain lipid metabolism, or of these fibroblasts under different conditions. PMID- 10822163 TI - Inhibition of striatal dopamine release by glycine and glycyldodecylamide. AB - Phencyclidine (PCP) and other N-methyl-D-aspartate (NMDA) antagonists induce schizophrenia-like symptoms in humans. In rodents, PCP induces a syndrome of stereotypies and hyperactivity that is accompanied by stimulation of striatal dopamine release. Glycine and other NMDA agonists reverse PCP-induced behaviors in rodents and ameliorate PCP psychosis-like symptoms of schizophrenia in clinical trials. Glycine levels in vivo are regulated by the actions of glycine (GLYT1) transporters. The present study investigates effects of glycine and the prototypic glycine transport inhibitor glycyldodecylamide (GDA) on striatal dopamine release in vitro using a mouse striatal assay. Glycine and GDA significantly inhibit NMDA-induced striatal dopamine release, consistent with their ability to enhance local striatal inhibitory neurotransmission in vitro and to reverse PCP-induced hyperactivity in vivo. PMID- 10822164 TI - An autoradiographic study of [3H]AMPA receptor binding and in situ hybridization of AMPA sensitive glutamate receptor A (GluR-A) subunits following morphine withdrawal in the rat brain. AB - Chronic treatment with opioids is well known to result in the development of physical dependence. More recently, glutamatergic mechanisms have been implicated in expression of the withdrawal syndrome from opioids. To better examine glutamatergic involvement, an autoradiographic study of [3H]AMPA receptor binding and an assessment of in situ hybridization of AMPA sensitive glutamate receptor A (GluR-A) subunits in the rat brain were each performed 7 h after withdrawal from morphine infusion. Animals were rendered dependent by intracerebroventricular (i.c.v.) infusion of morphine (26 nmol/microl/h) via osmotic minipumps for 3 days. Brain sections of 14-microm thickness were incubated with 15 nM [3H]AMPA for quantitation of binding to the AMPA receptor. The probe for in situ hybridization was labeled at its 3' end using terminal deoxynucleotidyl transferase and [35S]dATP. The highest degree of [3H]AMPA binding was shown in the hippocampus. The extent of [3H]AMPA binding was increased significantly in the cortex areas (18-21%), caudate-putamen (20%), and hippocampus (7-9%) of rats following withdrawal from morphine. The highest levels of mRNA for GluR-A, flop and flip subunits, were found in the dentate gyrus and in the CA3 region of the hippocampus, respectively. The levels of mRNA for the flop form of GluR-A were decreased in the CA3 of hippocampus (8%) of the rat brain. The levels of mRNA for the flip form of GluR-A were increased in the parietal cortex (7%) and the entorhinal cortex (8%). Increases in the binding of [3H]AMPA to its receptor may play an important role during withdrawal from morphine dependence. PMID- 10822165 TI - Brain interleukin-1beta and tumor necrosis factor-alpha are involved in lipopolysaccharide-induced delayed rectal allodynia in awake rats. AB - Recently, we have developed a model of delayed (12 h) increase in sensitivity (allodynia) to rectal distension (RD) induced by intraperitoneal lipopolysaccharide (LPS) in awake rats. Thus, we examined whether central interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) are involved in LPS response. Abdominal contractions (criterion of visceral pain) were recorded in rats equipped with intramuscular electrodes. RDs were performed at various times after pharmacological treatments. RD induced abdominal contractions from a threshold volume of distension of 0.8 ml. At lowest volume (0.4 ml), this number was significantly increased 12 h after LPS. Intracerebroventricular (i.c.v.) injection of IL-1 receptor antagonist, IL-1beta converting enzyme inhibitor or recombinant human TNF-alpha soluble receptor reduced LPS-induced increase of abdominal contractions at 0.4 ml volume of distension. When injected i.c.v., recombinant human IL-1beta and recombinant bovine TNF-alpha reproduced LPS response at 9 and 12 h and at 6 and 9 h, respectively. These data suggest that IL-1beta and TNF-alpha act centrally to induce delayed rectal hypersensitivity and that central release of these cytokines is responsible of LPS-induced delayed (12 h) rectal allodynia. PMID- 10822166 TI - Dehydroepiandrosterone sulfate (DHEAS) counteracts decremental effects of corticosterone on dentate gyrus LTP. Implications for depression. AB - It is well-established that levels of corticosterone sufficient to occupy Type II glucocorticoid receptors produce a decrement in long-term potentiation (LTP) in the dentate gyrus of the hippocampus in rats. In the present series of experiments we investigate the interaction of corticosterone and the neurosteroid dehydroepiandrosterone sulfate (DHEAS) on LTP in the rat dentate gyrus. In confirmation of previous studies, we found that corticosterone (2 mg/kg) had decremental effects on LTP. However, simultaneous injection of corticosterone and DHEAS (30 mg/kg) elicited excitatory post-synaptic potentials and population spikes that were not significantly different from those observed in control animals. The results are discussed in terms of the interaction of the two hormones, the agonist effects of DHEAS on sigma receptors, and their relation with the antidepressant effects of DHEA. PMID- 10822167 TI - Signal transduction of mucous secretion by bronchial gland cells. AB - Bronchial glands, which consist of mucous and serous cells, are abundant in human airways, playing a major role in the airway secretion. Cl(-) secretion is accompanied by water transport to the lumen in the acinar cells of bronchial glands. Agonists that increase [Ca(2+)]i induce the Cl(-) secretion in bronchial glands. Ca(2+) release from a IP(3)-sensitive Ca(2+) pool at the apical portion stimulates and opens Ca(2+)-sensitive Cl(-) channels at the apical membrane, producing Cl(-) secretion in bronchial glands. K(+) channels at the basolateral membranes are Ca(2+)-sensitive and activated by Ca(2+) release from a cADPribose sensitive Ca(2+) pool, maintaining the Cl(-) secretion in bronchial glands. Further, cADP ribose in concert with IP(3) induce [Ca(2+)]i oscillation, inducing Cl(-) secretion in bronchial glands. Some tyrosine kinases are involved in the Cl(-) secretion in bronchial glands. Mucous and serous cells in bronchial glands take part in mucin secretion and the secretion of defensive substances (glycoconjugates), respectively. [Ca(2+)]i oscillations are shown to play a central role in the exocytosis of secretory granules in serous cells of bronchial glands. Other signal transductions of mucin and glycoconjugates in airway gland cells remain to be studied, although agonists which increase [cAMP]i are also well known to induce mucin and glycoconjugate secretion from airway glands. PMID- 10822168 TI - Expression of human choline kinase in NIH 3T3 fibroblasts increases the mitogenic potential of insulin and insulin-like growth factor I. AB - In mammalian cells, growth factors, oncogenes, and carcinogens stimulate phosphocholine (PCho) synthesis by choline kinase (CK), suggesting that PCho may regulate cell growth. To validate the role of PCho in mitogenesis, we determined the effects of insulin, insulin-like growth factor I (IGF-I), and other growth factors on DNA synthesis in NIH 3T3 fibroblast sublines highly expressing human choline kinase (CK) without increasing phosphatidylcholine synthesis. In serum starved CK expressor cells, insulin and IGF-I stimulated DNA synthesis, p70 S6 kinase (p70 S6K) activity, phosphatidylinositol 3-kinase (PI3K) activity, and activating phosphorylation of p42/p44 mitogen-activated protein kinases (MAPK) to greater extents than in the corresponding vector control cells. Furthermore, the CK inhibitor hemicholinium-3 (HC-3) inhibited insulin- and IGF-I-induced DNA synthesis in the CK overexpressors, but not in the vector control cells. The results indicate that high cellular levels of PCho potentiate insulin- and IGF-I induced DNA synthesis by MAPK- and p70 S6K-regulated mechanisms. PMID- 10822169 TI - D609-phosphatidylcholine-specific phospholipase C inhibitor attenuates thapsigargin-induced sodium influx in human lymphocytes. AB - Previously, we reported that the phosphatidylcholine-specific phospholipase C (PC PLC) inhibitor tricyclodecan-9-yl xanthogenate (D609) potentiates thapsigargin induced Ca(2+) influx in human lymphocytes. In the present study we examined the effect of D609 on the thapsigargin-induced Na(+) entry. We found that the early phase of the thapsigargin-induced increase in the intracellular Na(+) concentration (approx. 1-2 min after stimulation) was attenuated after preincubation of lymphocytes with D609. By contrast, thapsigargin-induced Na(+) influx was not affected in the presence butan-1-ol, which inhibits phosphatidylcholine-specific phospholipase D (PC-PLD). The thapsigargin-induced Na(+) influx could be mimicked by PC-PLC exogenously added to the lymphocyte suspension, whereas addition of PC-PLD had no effect. In addition, thapsigargin stimulated formation of the physiological PC-PLC products, diacylglycerol. Cell permeable diacylglycerol analogue, dioctanoyl-glycerol (DOG), produced time- and concentration-dependent increase in the intracellular Na(+) concentration. Both thapsigargin- and DOG-induced Na(+) increases were not affected in the presence of Na(+)/H(+) antiport inhibitor, HOE609, or Na(+)/Ca(2+) antiport inhibitor, dimethylthiourea, as well as in the presence of Co(2+) and Ni(2+), which block store-operated Ca(2+) entry. By contrast, markedly reduced thapsigargin- and DOG induced Na(+) influx were noted in the presence of flufenamic acid, which blocks the non-selective cation current (I(CRANC)). In conclusion, our results suggest that diacylglycerol released due to the PC-PLC activation contributes to the thapsigargin-induced Na(+) entry. PMID- 10822170 TI - Protein kinase C-alpha and -beta play antagonistic roles in the differentiation process of THP-1 cells. AB - The roles of protein kinase C (PKC) isoenzymes in the differentiation process of THP-1 cells are investigated. Inhibition of PKC by RO 31-8220 reduces the phagocytosis of latex particles and the release of superoxide, prostaglandin E(2) (PGE(2)), and tumour necrosis factor (TNF)-alpha. The proliferation of THP-1 cells is slightly enhanced by RO 31-8220. Stable transfection of THP-1 cells with asPKC-alpha, and incubation of THP-1 cells with antisense (as) PKC-alpha oligodeoxynucleotides reduces PKC-alpha levels and PKC activity. asPKC-alpha transfected THP-1 cells show a decreased phagocytosis and a decreased release of superoxide, PGE(2) and TNF-alpha. The proliferation of asPKC-alpha-transfected THP-1 cells is enhanced. Stable transfection of THP-1 cells with asPKC-beta, and incubation of THP-1 cells with asPKC-beta oligodeoxynucleotides, reduces PKC-beta levels and PKC activity. asPKC-beta-transfected THP-1 cells show a decreased phagocytosis, a decreased TNF-alpha release, and a decreased proliferation. However, no difference is measured in the release of superoxide and PGE(2). These results suggest that: (1) PKC-alpha but not PKC-beta is involved in the release of superoxide and PGE(2); (2) TNF-alpha release and the phagocytosis of latex particles are mediated by PKC-alpha, PKC-beta, and other PKC isoenzymes; and (3) PKC-alpha and PKC-beta play antagonistic roles in the differentiation process of THP-1 cells. PKC-alpha promotes the differentiation process of THP-1 cells, PKC beta retards the differentiation of THP-1 cells into macrophage-like cells. PMID- 10822171 TI - The effects of agonist stimulation and beta(2)-adrenergic receptor level on cellular distribution of gs(alpha) protein. AB - This study examines the effects of adrenergic ligands, cholera toxin, forskolin, and varying levels of beta(2) adrenergic receptors (beta(2)AR) on the cellular distribution of Gs(alpha) subunits in CHO cells. Localization of Gs(alpha) was evaluated by confocal microscopy and beta(2)AR-mediated signalling was assessed by adenylyl cyclase (AC) activity. In cells expressing 0.2 pmol/mg protein beta(2)ARs (WT18), the localization of Gs(alpha) subunit was restricted to the plasma membrane region. Isoproterenol (ISO), cholera toxin or forskolin elicited redistribution of cellular Gs(alpha) so that Gs(alpha) appeared as intense spots throughout the plasma membrane as well as the cytoplasm. Exposure to a neutral beta(2)AR antagonist, alprenolol, prevented the ISO-stimulated Gs(alpha) translocation from peripheral to inner cytoplasm. In cells expressing high level of beta(2)ARs (8.2 pmol/mg) (WT4), basal and ISO-stimulated AC activities were significantly elevated when compared to the values detected in WT18 clone, suggesting a positive correlation between receptor expression and receptor mediated signalling. Basal Gs(alpha) distribution in this group was similar to that observed in ISO-, cholera toxin-, or forskolin-stimulated WT18 clone. ISO, cholera toxin, or forskolin did not change the distribution of Gs(alpha) significantly when tested in WT4 clone. No difference in the cellular level of Gs(alpha) protein between WT18 and WT4 clones was detected. Alprenolol did not affect the distribution of Gs(alpha) in WT4 clone. ICI 118,551, a negative beta(2)AR antagonist, altered Gs(alpha) distribution from a dispersed basal pattern to a membrane-confined pattern. The latter appearance was similar to that observed in unstimulated WT18 clone. Taken together, these data suggest that: (1) enhanced beta(2)AR-Gs(alpha) coupling induced by agonist stimulation or by increased expression of beta(2)ARs remodel the cellular distribution of Gs(alpha); (2) the alteration in Gs(alpha) distribution induced by beta(2)AR overexpression provides evidence for agonist-independent interaction of beta(2)AR and Gs(alpha), that can be inhibited by a negative antagonist but not by a neutral antagonist; and (3) forskolin influences the activity state of Gs(alpha) that displays a Gs(alpha) distribution pattern comparable to that observed when Gs(alpha) is activated via beta(2)AR stimulation or directly by cholera toxin. PMID- 10822172 TI - Effect of flutamide-induced androgen-receptor blockade on adenylate cyclase activation through G-protein coupled receptors in rat prostate. AB - The effect of the antiandrogen flutamide on the prostatic vasoactive intestinal peptide (VIP) receptor/effector system was studied in rats. Rats were s.c. injected with a daily dose of flutamide (15 mg/kg B.W.) or vehicle for 14 days. Drug treatment resulted in histological evidence of gland involution and increased plasma membrane fluidity as estimated by fluorescence spectroscopy. The number of VIP receptors and the stimulatory effect of VIP on adenylate cyclase activity in prostatic membranes decreased in flutamide-treated rats. However, the pattern of forskolin stimulation of the enzyme activity was not modified by this drug. Androgen-receptor blockade by flutamide also decreased the prostatic levels of alpha(s,) alpha(i1/2), and alpha(i3/0) G-protein subunits, as estimated by an immunological procedure. Whereas apoptotic DNA fragmentation was evidenced in prostate from 3-day castrated animals, a heterogeneous electrophoretic pattern was observed after flutamide treatment. Thus, androgen-receptor blockade by flutamide results in an important impairment of the components of the VIP receptor/effector system in rat prostate as well as in a modification of their coupling extent, which is presumably due to differences observed in plasma membrane fluidity. These results represent a crosstalk in the prostate between two mechanisms of signal transduction involved in cell proliferation. PMID- 10822173 TI - The phosphatidylinositol polyphosphate 5-phosphatase SHIP1 associates with the dok1 phosphoprotein in bcr-Abl transformed cells. AB - The initial phase of chronic myelogenous leukemia (CML) is triggered by constitutive protein tyrosine kinase activity of the chimeric kinase p210(bcr abl) (Bcr-Abl). A major substrate of Bcr-Abl was recently identified as the RasGAP-associated 62 kDa docking protein Dok1. Here, we report complex formation between endogenous Dok1 and the SH2 domain-containing phosphatidylinositol polyphosphate 5-phosphatase SHIP1 in hematopoietic cells expressing Bcr-Abl. Expression of Bcr-Abl induced tyrosine phosphorylation of both Dok1 and SHIP1 and the formation of a Dok1/SHIP1 complex. Tyr(P) SHIP1 was also bound to Shc in Bcr Abl expressing cells. A small amount of Shc/SHIP1/Dok1 trimolecular complex was detected and this was due to binding of Dok1 to SHIP1 that was bound to Shc. In contrast, association of Dok1 with SHIP1 or RasGAP was mutually exclusive. Both the SH2 domain of SHIP1 and the PTB domain of Dok1 were required for complex formation between the two proteins. Neither the specific activity of SHIP1 as an inositol phosphate 5-phosphatase nor the subcellular localization of SHIP1 appeared to be altered by tyrosine phosphorylation. However, the Dok1/SHIP1 complex was only detected in the cytosolic fraction of Bcr-Abl transformed hematopoietic cells. We propose that interaction between Dok1 and SHIP1 modulates the ability of these two proteins to interact with other cytosolic binding partners. PMID- 10822174 TI - A peptide of the alpha 3(IV) chain of type IV collagen modulates stimulated neutrophil function via activation of cAMP-dependent protein kinase and Ser/Thr protein phosphatase. AB - Previous reports from our laboratories showed that type IV collagen from anterior lens capsule (ALC) inhibited stimulated neutrophil function. This property was shown to reside in the region comprising residues 185-203 of the non-collagenous domain (NC1) of the alpha 3(IV) chain. We also reported that ALC-type IV collagen or the synthetic alpha 3(IV) 185-203 peptide, induced a rise in intracellular cAMP which persisted for up to 60 minutes. In the present work we extend our previous studies on signal transduction by alpha 3(IV) 185-203 and we provide new data showing the involvement of cAMP-dependent PKA and protein phosphatases. The data also show that the alpha 3(IV) peptide triggered a rise in intracellular calcium that was dependent on phospholipase C activation. Inhibitors of the Ca(2+)/calmodulin system suppressed both the alpha 3(IV) 185-203 peptide-induced cAMP increase and the inhibitory activity of the peptide on f-Met-Leu-Phe triggered O(2)(-) generation. When alpha 3(IV) 185-203 peptide-induced calcium mobilization was blocked by U-73122, an inhibitor of phospholipase C activation, or by BAPTA/AM, a chelator of intracellular calcium, the inhibitory effect of the peptide on PMA-triggered O(2)(-) production was also abolished. These findings provide evidence that signal transduction by the alpha 3(IV) peptide occurs via pathways which involve calcium. Indeed, the cAMP increase was shown to be mediated by adenosine and adenosine A2 receptors and required calcium elevation, since adenosine deaminase, theophilline, dimethylpropargylxanthine, trifluoperazine or autocamtide-2 related inhibitory peptide, suppressed the activity of the alpha 3(IV) peptide. The inhibitory effect of the peptide on f Met-Leu-Phe-induced O(2)(-) generation was slightly affected by 1 microM KT5720 or H89, two inhibitors of cAMP-dependent PKA, but was completely suppressed by 10 nM calyculin A or 10 microM okadaic acid, two inhibitors of ser/thr phosphatases. These results suggest that Ser/Thr protein phosphatases and/or cAMP-dependent PKA are involved in signal transduction by the alpha 3(IV) 185-203 peptide and is consistent with the concept that adenosine receptor occupancy modulates neutrophil function. PMID- 10822175 TI - Nuclear targeting of proteins: how many different signals? AB - The nuclear import of proteins into the cell nucleus involves the recognition of a nuclear localization signal sequence, borne by the protein to be transported, by complex molecules called importins, that will subsequently mediate the crossing over of the nuclear envelope. The most frequently encountered signal sequence is made up of short stretches of basic amino acid residues and is recognized by importins alpha and/or beta. Other signal sequences have been described, and some have been shown to mediate the association with importins other than importin alpha or beta. Recently, approaches have been developed that allow the cloning, on a functional basis, of sequences able to specify the nuclear localization of proteins. A variety of peptidic motifs of limited size which do not contain previously described signal sequences were isolated in such assays. It reveals that the spectrum of sequences that are able to target a protein to the cell nucleus may be wider than currently expected. It will probably also lead to the identification of novel target sequences for importins and will demonstrate the implication of additional members of this family of proteins in nuclear transport. PMID- 10822176 TI - Functional compartments in rat mast cells for cAMP and calcium on histamine release. AB - The crosstalk between 3', 5'-cyclic adenosine monophosphate (cAMP), intracellular calcium, and histamine release in rat mast cells using the stimulatory effect of three different drugs, thapsigargin, sodium fluoride (NaF), and compound 48/80 were studied. Each of these drugs induces histamine release by different mechanisms. The transducting pathways modulating cAMP and intracellular calcium levels were modified by using, cholera toxin (CTX) which ADP-rybosylates Gs protein, pertussis toxin (PTX) which ADP-rybosylates Gi-protein, and okadaic acid (OA) which inhibits phosphatases 1 and 2a. Our results show that CTX increased cAMP levels and inhibited histamine release elicited by thapsigargin and compound 48/80. The inhibitory effect of CTX on histamine release was potentiated by OA in the presence of compound 48/80 but was decreased in the presence of thapsigargin. Calcium uptake was stimulated by NaF and compound 48/80. The previous treatment with OA increased calcium uptake when combined with compound 48/80 but not with NaF. Treatment with NaF highly stimulated calcium uptake and cAMP levels only when combined with OA and CTX. These results suggest that the modulatory effect of intracellular calcium and cAMP on histamine release depend more on the crosstalk of the activated signal transducting pathway than on the final level of calcium or cAMP, further supporting the theory that rat mast cells are divided into functionally distinct compartments. PMID- 10822177 TI - Expanding the scope of HIV prevention for adolescents: beyond individual-level interventions. PMID- 10822178 TI - Compliance in adolescents with chronic diseases: a review. PMID- 10822179 TI - Confidential billing accounts for adolescents in private practice. AB - The purpose of this study was to determine how individual confidential billing accounts for adolescents in private practice would be received, and to assess the impact of these accounts on practice reimbursement. Adolescents wishing to keep confidential any part of the charges for an office visit were offered individual billing accounts. Enrollees agreed to pay whenever and whatever they could. One attempt was made to contact nonpayers at least 3 months after starting their accounts. The mean charge for 40 confidential accounts was $42 (total charges $1489) and all of the charges were for laboratory testing. Participants reimbursed 38% of the total charged ($565/$1489). Confidential accounts were well received by study participants and may improve access to confidential care. PMID- 10822180 TI - High-risk adolescents and female condoms: knowledge, attitudes, and use patterns. AB - PURPOSE: To explore data on high-risk male and female adolescents' attitudes towards female condoms, compared with male condoms. METHODS: Exploratory survey research was utilized with a convenience sample of 65 high-risk adolescents at an emergency homeless shelter. A peer-led intervention was conducted and pre-test and post-test interviews explored barriers to female condom use. The intervention consisted of 15- to 30-minute small group sessions, discussing female condoms' construction; purpose of the rings; efficacy preventing pregnancy and sexually transmitted infection (STIs); and how to lubricate, insert, and use. Content and Chi-square analyses were utilized. RESULTS: Sixty-three percent used male condoms as their primary contraceptive method; almost half (48%) said they always used male condoms, but 44% reported having sex without a male condom at least once in the 2 weeks prior to pre-test. Ninety-five percent had heard of the female condom, half 'good' and 24% 'bad' things, but only 15% had ever used one. At post test all respondents gave reasons they might use female condoms in the future, and 77% gave reasons why they might not. Most (73%) adolescents said they would still prefer the male to the female condom. The major potential barriers to adolescents' female condom use were not having female condoms available and/or females feeling uncomfortable inserting them. CONCLUSIONS: Female condoms should be offered to adolescents as an additional choice rather than as replacements for male condoms. Further research is needed to assure access, availability, and comfort with female condoms and male participation in their use. PMID- 10822181 TI - African-American and Latina adolescent mothers' infant feeding decisions and breastfeeding practices: a qualitative study. AB - PURPOSE: To explore minority teen mothers' perceptions of breastfeeding and the influences on infant feeding choices. METHODS: A qualitative study using semistructured ethnographic interviews and focus groups involving 35 Latina and African-American girls in Chicago between the ages of 12 and 19 years who were primiparous and were currently pregnant or had delivered within the past 3 months. RESULTS: Adolescents identified three main influences on infant feeding decisions and practices: (a) their perceptions of the benefits of breastfeeding, (b) their perceptions of the problems with breastfeeding, and (c) influential people. In this study, teens reported no single influence which determined infant feeding choices. The decision to breastfeed was a dynamic process. Teens recognized that breastfeeding offered many benefits including facilitating maternal-child bonding and promoting the baby's health, but concern was raised regarding a potential for excessive attachment between teen mother and baby. Fear of pain, embarrassment with public exposure, and unease with the act of breastfeeding acted as barriers for teenagers who were considering breastfeeding. Teenagers discussed the breast pump as a strategy in dealing with these barriers. The adolescents' mothers continued to be an important influence. CONCLUSIONS: The ranges of perceptions and influences that minority adolescent mothers have identified as affecting their infant feeding choices, illustrated and explained in the teens' own words, are helpful to health care providers as they counsel teen mothers about infant feeding options. PMID- 10822182 TI - The timing of norplant insertion and postpartum depression in teenagers. AB - BACKGROUND: This study tested the hypothesis that teenagers who have Norplant inserted during the puerperium report more depressive symptoms during the first postpartum year than their peers who do not receive Norplant. METHODS: We studied the prevalence of depressive symptoms in a group of 212 mothers aged 19 years less, in relation to the timing of Norplant insertion. The participants were divided into 3 groups: 100 (47%) had Norplant inserted during the puerperium (early Norplant users); 72 (34%) had Norplant inserted during the next 10 months (late Norplant users); and 40 (19%) used other contraceptives (40% oral contraceptives; 17% condoms; 43% nothing). Depressive symptoms were measured with the Center for Epidemiologic Studies - Depression Scale. Postpartum depression was defined as a scale score >/=16, 6-12 months after Norplant insertion or delivery. Variables examined as potential confounders were identified a priori from a review of the literature and controlled for in analysis of variance. RESULTS: At delivery, members of the 3 contraceptive groups did not differ significantly with regard to age, race, parity, educational, marital, or socioeconomic status. Late Norplant users were, however, more apt to have new boyfriends (p =. 03), to rate the support they received from the baby's father as poor (p =.004), and experience depression prior to Norplant insertion (p =.02). Contrary to the study hypothesis, late rather than early Norplant insertion was associated with postpartum depression. Multivariate analyses identified 3 independent predictors of the severity of depressive symptoms at follow-up (depression prior to Norplant insertion, a new boyfriend at delivery, and late Norplant insertion); R(2) = 41.3%. CONCLUSIONS: Contrary to the study hypothesis, puerperal Norplant insertion did not exacerbate postpartum depression. Delaying Norplant insertion may increase the risk of depression during the first postpartum year, particularly in teenagers with other psychosocial risk factors. PMID- 10822183 TI - Influence of parental height and sociodemographic factors on adolescent height in Brazil. AB - OBJECTIVE: To analyze the influence of parental stature and environmental factors on the stature of adolescents from a national survey sample. METHODS: A nationwide survey was carried out in 1989 among a stratified, two-stage, probability cluster sample of 14,455 Brazilian households to provide estimates of anthropometric deficits for urban and rural populations from the five regions of the country. Stature was measured for 5681 boys and girls age 14-18 years, 78.9% of their fathers, and 93.8% of their mothers. Associations between explanatory variables and adolescent height in centimeters were assessed by fitting multiple linear models to the data. RESULTS: The predicted effects of parental stature and environmental conditions together sum to a total of 17 cm when comparing a boy born to parents with stature below the median and living in the underdeveloped rural Northeast region (1.56 m) with one born to parents with stature above the median and living in the partially industrialized urban South region (1.73 m). For girls, this estimated difference was 12 cm. For boys, the overall influence of parents' stature was 10 cm (R(2)= 0.40) and the sociodemographic factors had an overall influence of 7 cm (R(2) = 0.29). For girls, these values were 7 cm (R(2)= 0.35) for the parental influence and 5 cm (R(2) = 0.11) for the sociodemographic factors. CONCLUSIONS: Mother's stature had the same influence on adolescent's stature as father's stature. Independent of parental stature, environmental factors have a strong influence on adolescent stature, particularly among boys. PMID- 10822184 TI - Protective correlates of stages in adolescent substance use: a Swiss National Study. AB - OBJECTIVE: To identify factors potentially protective against involvement with additional illicit drugs among adolescent users of marijuana. DESIGN AND METHODS: A cross-sectional, secondary analysis of a national data set of Swiss adolescents was performed. The original survey was conducted in 1992-1993 (N= 9268). Stages of substance use were conceptualized as lifetime use of marijuana of three or more times and as use of marijuana and other illicit drugs (opiates, inhalants, cocaine, hallucinogens, or stimulants) more than twice in the past month. Behavioral characteristics associated with level of drug use were examined. Based on the resilience literature, factors potentially protective against progression of drug use were examined in their bivariate relationships and analyzed in a multiple regression model. RESULTS: Almost one quarter of the surveyed adolescents were classified as past or current marijuana users. One in 20 of these adolescents also used other illicit drugs. Prevalence of risk-related behaviors (e.g., antisocial behavior, accidents, suicide attempts) was elevated for marijuana users compared to nonusers and even more elevated for users of marijuana with other illicit drugs. In a multivariate logistic regression model, communicating well with a parent was significantly associated with not having progressed to use of illicit drugs other than marijuana [for males, odds ratio (OR) = 0.72, confidence interval (CI) = 0.55, 0.92; for females, OR = 0.60, CI = 0.39, 0.93]. Other significant protective correlates for boys were academic achievement (OR = 0.66, CI = 0.47, 0.92), type of education: school versus apprenticeship (OR = 0.42, CI = 0.21, 0.87), confiding in a family member (OR = 0.49, CI = 0.28, 0.85), and regular participation in a sports club (OR = 0.39, CI = 0.22, 0.77). CONCLUSION: Within this national sample of Swiss adolescents, a cross-sectional analysis identified the quality of relationships within the family and factors related to school associated with lack of progression of illicit substance use. However, longitudinal studies will be necessary to confirm these associations and to allow for designing interventions targeting the enhancement of protective factors among young people already at risk for serious substance use. PMID- 10822185 TI - Fetal arrhythmias. AB - Fetal arrhythmias may be benign or life-threatening. Benign disturbances in fetal cardiac rhythm are relatively common, and their clinical manifestations are reviewed. Life-threatening fetal arrhythmias include supraventricular tachycardias, atrial flutter, ventricular or junctional tachycardia, chaotic atrial tachycardia, and bradyarrhythmias such as second or third degree AV block. The incidence, diagnostic characteristics and treatment of each of these are reviewed. In addition, the pathophysiology of hydrops fetalis, embryology of the conduction system and transplacental drug transfer characteristics are reviewed. PMID- 10822186 TI - Neonatal bradycardia. AB - Heart rate monitoring has become a ubiquitous part of fetal and neonatal assessment, and has made detection of bradycardia in the fetal and neonatal periods a frequent occurrence. Evaluation of a fetus or neonate with bradycardia requires an understanding of the mechanisms of bradycardia as well as the cardiac and non-cardiac causes of bradycardia. The mechanisms of bradycardia include sinus bradycardia, abnormalities of sinus node function and abnormalities of atrioventricular conduction. In the instances where sinus bradycardia is pathologic, it usually results from non-cardiac disease. Sinus node dysfunction is rare early in life but can arise from surgical interventions, congenital heart disease, or endovascular manipulations. Abnormalities of atrioventricular conduction have a similar etiology but are more common than sinus node disease. Atrioventricular nodal disease can also result from maternal collagen vascular disease, even in the absence of symptoms in the mother. In these cases, epidemiological issues such as heart block in subsequent pregnancies and the maternal risk of developing symptomatic collagen vascular disease become important. The approach to treatment and long-term prognosis for bradycardia in the neonate is highly dependent on the underlying etiology and on the presence of concurrent factors such as structural heart disease. PMID- 10822187 TI - Supraventricular tachycardia in the neonate and infant. AB - Supraventricular tachycardia (SVT) is the most common sustained arrhythmia to present in the neonatal and infancy age group. Predisposing factors (congenital heart disease, drug administration, illness and fever) occur only in 15% of infants. The presentation of SVT in the neonate is frequently subtle, and may include pallor, cyanosis, restlessness, irritability, feeding difficulty, tachypnea, diaphoresis and grunting. Congestive heart failure is more common in infants under 4 months of age (35% incidence). Age-related differences in the distribution of SVT mechanisms occur in different age groups. In infants under 1 year of age, the mechanisms underlying SVT are atrial tachycardia (15%), AV nodal re-entry tachycardia (5%), and AV reciprocating tachycardia (80%). Options for acute management include: use of the diving reflex, intravenous adenosine, transesophageal pacing, and cardioversion. Intravenous administration of verapamil should be avoided. Data regarding freedom from recurrence of untreated SVT in the first year of life are limited, and may be in the range of 25-60%. Chronic therapy with digoxin, beta-blockers, flecainide, sotalol and amiodarone has proved effective in controlling recurrent episodes of SVT. Radiofrequency ablation can be employed successfully in medically refractory cases, but should be avoided in this age group (increased complication rate). PMID- 10822188 TI - Ventricular arrhythmias. AB - A spectrum of distinctive clinical presentations and electrocardiographic patterns have been recognized in neonates with ventricular arrhythmias. These may range from an incidental finding on a routine physical to cardiovascular collapse due to ventricular fibrillation. It has become increasingly important that the clinician considers ventricular tachycardia in the neonate with tachycardia when the QRS normal does not appear normal. In general, isolated premature ventricular depolarizations, couplets and non-sustained ventricular tachycardia in the absence of heart disease are associated with a favorable prognosis. Most of these arrhythmias tend to resolve during the first month of life. Conversely, sustained ventricular arrhythmias associated with ischemia, myocarditis or ventricular tumors are associated with a guarded prognosis. Treatment is based on the definition of associated cardiovascular disease, support of hemodynamic status and the judicious use of antiarrhythmic agents. Finally, there has been an increased recognition of idiopathic forms of ventricular tachycardia in the neonate which are associated with a favorable prognosis and may not require pharmacologic treatment. This review will discuss these arrhythmias in neonates, associated forms of cardiovascular disease, current treatment options and long term prognosis. PMID- 10822189 TI - Neonatal long QT syndrome and sudden cardiac death. AB - Neonatal sudden cardiac death most often results from cardiac electrical diseases, cardiomyopathies, or sudden infant death syndrome. In infants without a known premortem diagnosis or abnormalities identified at autopsy, sudden infant death syndrome accounts for the vast majority of sudden deaths. Potential cardiac causes of some sudden infant death syndrome cases may include malignant brady- or tachyarrhythmias and congenital long QT syndrome. The possible mechanisms include abnormal brain stem respiratory control of arousal, dysautonomia and malignant cardiac bradyarrhythmias or tachyarrhythmias. Screening for neonatal sudden cardiac death may not be feasible, but hopefully through careful review of history, physical examination, and family health history, and judicious diagnostic testing, can the risk of cardiac sudden death be reduced. Further comprehension of the genetic basis of inherited arrhythmia disorders may help elucidate the mechanisms of arrhythmogenesis and etiologies of sudden infant death. Prevention and treatment of these disorders may also be improved through more detailed understanding of the molecular basis of cardiac electrical pathophysiology. PMID- 10822190 TI - Antiarrhythmic drug therapy in the neonate. AB - The neonate presents a challenge for the practitioner considering antiarrhythmic therapy: pharmacokinetics are different than in older children or adults; and the arrhythmia substrate may also differ, with respect to issues of ion channel and autonomic nervous system development. This paper reviews the need for antiarrhythmic drug therapy in the neonate, developmental aspects of pharmacokinetics, autonomic regulation and cellular electrophysiology and the antiarrhythmics available today with an emphasis on newer drug therapy. PMID- 10822191 TI - Applications of pacing strategies in neonates and infants. AB - Pacing in neonates and infants continues to be challenging due to size constraints, growth potential, and the need for life long pacing. Indications for permanent pacing in pediatric patients have been difficult to determine due to the lack of data from controlled studies and multicenter trials. Temporary pacing has been useful to restore cardiac output in pediatric patients quickly and efficiently. Methods of temporary pacing include transcutaneous, transesophageal, transvenous, and epicardial. Permanent pacemaker implantation can be accomplished by transvenous or epicardial approaches, but the use of transvenous pacing in neonates and infants offers no advantages over epicardial pacing. Transvenous pacing in infants is often prohibitive due to size and growth constraints as well as the subsequent risk of skin erosion and venous thrombosis. Smaller pulse generators, multiprogrammable features, and steroid-eluting epicardial leads are a few of the technological advances that have made pacing in neonates and infants easier and safer. Data supporting the use of pacing systems in very young patients are sparse. Pacing 'indications' should be viewed as guidelines until such data can be accumulated. PMID- 10822192 TI - Radiofrequency catheter ablation of arrhythmias in infants and small children. AB - The issue of the proper use of radiofrequency catheter ablation in infants and small children is controversial. Rarely, catheter ablation techniques are indicated to cure malignant arrhythmias in the pediatric population. Indications, techniques, complications and success rates are reviewed. PMID- 10822207 TI - Q&A about this issue PMID- 10822208 TI - Multiple drug allergy syndrome. PMID- 10822209 TI - The canadian society for clinical pharmacology in the post-royal college era PMID- 10822210 TI - Irbesartan: review of pharmacology and comparative properties. AB - The recently developed and marketed angiotensin II type 1 (AT1) receptor blockers (ARBs) have demonstrated efficacy equivalent to that of other leading classes of antihypertensive agents, with superior tolerability profiles. The specific targeting of the AT1 receptor afforded by these agents has demonstrated more complete blockade of the renin-angiotensin system than that offered by angiotensin-converting enzyme inhibitors. These data notwithstanding, differentiation within the class of ARBs has been limited. With the accumulation of additional data with ARBs, it has recently become possible to make within class distinctions, based in large part on the individual pharmacological profiles of the ARBs. To this end, absorption, distribution, half-life, dose response and level of angiotensin II antagonism are of special note. When these properties are viewed as a group, the ARB irbesartan appears to offer advantages beyond those attained with other ARBs. Irbesartan is well absorbed, does not require biotransformation to an active metabolite to exert its antihypertensive activity, offers a large volume of distribution, has a half-life that is sufficient to allow once-daily dosing, is associated with a strong and consistent dose-response and has been demonstrated to provide a level of angiotensin II antagonism that is statistically superior to that offered by some other ARBs. These pharmacological differences may explain the clinical superiority of irbesartan compared with losartan, the first member of the ARB class. As even more data on the ARBs become available, the ability to determine the advantages of specific members of this class will be enhanced, distinctions that already have begun to come to light. PMID- 10822211 TI - Effects of indapamide versus hydrochlorothiazide on plasma lipids and lipoproteins in hypertensive patients: a direct comparison. AB - To test the claim that indapamide causes fewer adverse metabolic effects than hydrochlorothiazide (HCTZ), six months of treatment with HCTZ 25 mg/day was compared with six months of treatment with indapamide 2.5 mg in a randomized double-blind study of 44 patients with mild to moderate hypertension. Both drugs significantly reduced blood pressure, with no significant differences in the reduction of potassium and chloride; neither drug was associated with a significant change in plasma total cholesterol, high density lipoprotein (HDL), apolipoprotein A1, apolipoprotein B or the ratio of total cholesterol to HDL levels. Triglyceride levels increased significantly more with indapamide than with HCTZ (P=0.02). The two drugs had similar effects on blood pressure and serum potassium; neither drug affected plasma glucose. Claims that indapamide is superior to HCTZ appear unwarranted; in view of the substantial cost difference, this finding, if confirmed in larger studies, has important policy implications. PMID- 10822212 TI - Multiple antibiotic sensitivity syndrome in children. AB - BACKGROUND: Multiple antibiotic sensitivity syndrome with adverse drug reactions to multiple classes of antibiotics has been described in adults but is not well characterized in children. PATIENTS AND METHODS: Charts of children referred to the adverse drug reaction clinic at the Children's Hospital of Western Ontario, London, Ontario, with adverse drug reactions to multiple antibiotics were reviewed to determine the number of patients with adverse drug reactions to multiple classes of antibiotics and the clinical characteristics of the adverse events. RESULTS: The records of 97 children who were selected as possible candidates for multiple antibiotic sensitivity were studied. These records constituted 11% of referrals to a highly specialized adverse drug reaction clinic, suggesting that in usual clinical practice, this entity, if it does indeed constitute a distinct clinical entity, is quite uncommon. Age at time of the first adverse drug reaction was 26.1+/-26.3 (mean +/- SD) months. Among the 97 children, adverse reactions to five classes of antibiotic were noted in 3.1%, to four in 10.3%, to three in 47. 4% and to two in 39.2%. Most children (85.6%) experienced an adverse reaction to a penicillin, while 71.1% reacted to a cephalosporin, 80. 4% to a sulphonamide and 35.1% to a macrolide. Clinical presentations of the adverse reactions included urticaria or pruritus, other rash, serum sickness-like reaction, angioedema or anaphylaxis, erythema multiforme or Stevens-Johnson syndrome. CONCLUSIONS: There are children who have what appears to be immunologically mediated adverse drug reactions to antibiotics of multiple classes. These reactions, which most commonly manifest as urticaria or other rashes, follow drug use patterns. It remains to be defined whether this is a distinct clinical syndrome or a manifestation of a more fundamental problem in dealing with xenobiotics in the setting of infection. Further work on the immunological and/or biochemical determinants of the multiple antibiotic sensitivity syndrome (MASS) is needed to understand the pathophysiology and determinants of MASS and whether MASS constitutes a distinct clinical entity. PMID- 10822213 TI - Attitudes and practices of physicians and naturopaths toward herbal products, including use during pregnancy and lactation. AB - BACKGROUND: The popularity of complementary therapies continues to grow, and physicians are asked increasingly by their patients for information regarding these modalities. PURPOSE: To assess the impact of these trends on physicians and medical students, and compare their attitudes and practices with those of the naturopaths and their students, with particular interest in the use of herbal products during pregnancy and breastfeeding. MATERIALS AND METHODS: A detailed questionnaire was distributed by a medical student and a naturopathic student to a randomly selected group of physicians, medical students, naturopaths and naturopathic students. They were asked a variety of questions about their background, attitudes and practices concerning herbal products. RESULTS: Thirty eight per cent of the questionnaires were returned, with a total of 242 respondents. Fifty-four per cent of physicians discussed complementary therapies with their patients, whereas 100% of naturopaths discussed conventional medicines with their patients. The most popular product recommended by both medical doctors and naturopaths was echinacea, followed by St John's Wort. Eighty-six per cent of physicians, 74% of medical students, 66% of naturopaths and 50% of naturopathic students think that complementary medical education should be incorporated into the standard medical curriculum. Only one physician actually recommended a herbal product to a pregnant patient compared with 49% of the naturopaths who felt comfortable doing so. CONCLUSIONS: Complementary medicine has become a reality, and physicians are recommending herbal products to their patients, although on a smaller scale than are naturopaths. However, the two most popular herbal products are the same in each group. Physicians are less likely to recommend herbal products to pregnant and breastfeeding women than are naturopaths. PMID- 10822214 TI - Genesis of the normal and abnormal arterial pulse. PMID- 10822215 TI - Markers of bone turnover in the evaluation of the response to GH treatment in GH deficient children. PMID- 10822216 TI - Dynamics of bone turnover in children with GH deficiency treated with GH until final height. AB - OBJECTIVE: To examine the dynamics of bone turnover in children with growth hormone deficiency (GHD) during long-term treatment. DESIGN: We longitudinally measured growth velocity and serum concentrations of osteocalcin (OC), carboxyterminal propeptide of type I procollagen (PICP), and cross-linked carboxyterminal telopeptide of type I collagen (ICTP) in 24 patients with GHD during long-term GH treatment until final height (age: 7.7+/-0.7 and 16.9+/-0.5 years at baseline and at final height respectively). RESULTS: At baseline, OC, PICP, and ICTP levels were significantly (P<0.0001) reduced in comparison with prepubertal bone age-matched controls (10.2+/-2.3 microgram/l and 22.5+/-7.6 microgram/l; 187.8+/-26.2 microgram/l and 328. 4+/-74.3 microgram/l; 7.7+/-2.0 microgram/l and 14.2+/-1.3 microgram/l respectively). During the first year of treatment mean levels of the bone markers increased significantly (P<0.0001) with a peak at 12 months. After the first year of treatment, OC and PICP levels progressively declined, whereas ICTP levels remained stable until the final height; in any case, bone marker levels remained significantly higher (P<0.03 P<0.0001) than baseline. The change in bone marker levels at 6 and 12 months of treatment with respect to the baseline values was not related to growth rate during long-term treatment or final height. CONCLUSIONS: The results show that children with GHD have reduced bone turnover at baseline, and that long-term GH treatment is associated with a stimulation of bone turnover. OC, PICP, and ICTP do not predict growth rate during long-term treatment or final height in children with GHD. PMID- 10822217 TI - Familial growth hormone deficiency with mutated GHRH receptor gene: clinical and hormonal findings in homozygous and heterozygous individuals from Itabaianinha. AB - OBJECTIVE: To characterize clinically and hormonally the syndrome of autosomal recessive familial growth hormone deficiency (FGHD) recently identified in Itabaianinha, Sergipe, Brazil, caused by a novel mutation (mt) that inactivates the growth hormone-releasing hormone receptor (GHRH-R) gene. DESIGN: Clinical and hormonal evaluations were performed in 21 FGHD individuals (mt/mt group) aged 8 to 63 years, 13 heterozygotes for the GHRH-R mutation (wt/mt group) and 5 homozygotes for the wild type (wt) allele (wt/wt group), identified by genotyping of peripheral blood leukocyte DNA. METHODS: Clinical and hormonal characterization included physical examination and measurement of GH, IGF-I, IGF binding protein-3 (IGFBP-3), cortisol, prolactin, LH, FSH, and free thyroxine (FT4). RESULTS: Clinical features were consistent with isolated growth hormone deficiency. Height was significantly reduced in the mt/mt group compared with the wt/mt group (mean height standard deviation score (SDS) +/- s.d.: -7.35+/-1.37 vs -1.84+/-1.44 respectively, P<0. 0001), and the wt/wt group (-1.85 +/- 0.81, P=0.0007). The height of the 13 wt/mt subjects did not differ from the 5wt/wt individuals. Serum GH, IGF-I, IGF-I SDS, IGFBP-3 and IGFBP-3 SDS were all significantly lower in the mt/mt group than in the wt/mt and wt/wt groups. Two affected children treated with GH for 1 year showed a normal growth response. Serum IGF-I and IGF-I SDS were lower in wt/mt compared with wt/wt group, but did not reach statistical significance. IGF-I and IGF-I SDS correlated inversely with age in wt/mt group. CONCLUSIONS: FGHD due to an autosomal recessive GHRH-R gene mutation leads to marked dwarfism, phenotypically and hormonally indistinguishable from other forms of isolated GH deficiency. Heterozygotes for the GHRH-R mutation appear to have a partial defect in the GH/IGF axis, with no apparent height impairment. PMID- 10822218 TI - Variable growth hormone profiles following withdrawal of long-term 30mg slow release lanreotide treatment in acromegalic patients: clinical implications. AB - OBJECTIVE: Intramuscular injections of 30mg slow-release (SR) lanreotide (every 10 to 14 days) are an effective treatment in acromegalic patients. Because of an ongoing need to assess the efficacy and the tolerance of a new formulation of a depot preparation of lanreotide, we have evaluated prospectively GH profiles following withdrawal of 30mg slow-release lanreotide in a cohort of acromegalic patients. PATIENTS: Fifty-one acromegalic patients, controlled during long-term 30mg SR lanreotide treatment (GH: 1.44 +/- 0.64 microgram/l, IGF-I: 316 +/- 145ng/ml) (mean +/- s.d.), were studied following the withdrawal of the drug. MEASUREMENTS: Mean GH (half-hour samples, 0800-1200h), IGF-I and lanreotide levels were evaluated 14, 28, and 42 days following the last 30mg SR lanreotide injection. RESULTS: Mean GH levels remained below 2.5 microgram/l in 32 patients (group 1) twenty-eight days following SR lanreotide withdrawal. In these patients, mean GH and IGF-I levels had increased from 1.2 +/- 0.6 to 1.7 +/- 0.5 microgram/l (P < 0001), and from 283 +/- 138 to 359 +/- 168ng/ml (P < 0.001) respectively. In the 19 other patients (group 2), mean GH concentrations had risen above 2.5 microgram/l at 28 days following SR lanreotide withdrawal. Mean GH and IGF-I levels had increased from 1.9 +/- 0.4 to 5.1 +/- 2.8 microgram/l (P < 0.001), and from 371 +/- 143 to 568 +/- 206ng/ml (P < 0.001) respectively. Patients of groups 1 and 2 were comparable with regard to age, sex, tumoral status, mean GH levels before somatostatin analogue treatment, and previous treatments such as radiotherapy and duration of somatostatin analogue therapy, but 75% of group 1 patients underwent surgery compared with 37% of group 2 patients (P < 0.01). Twenty-eight days following SR lanreotide withdrawal, mean lanreotide levels in group 1 and group 2 had decreased from 1.6 +/- 0.7 to 0.6 +/ 0.3ng/ml (P < 0.001), and from 2.7 +/- 2.0 to 0.7 +/- 0.7ng/ml (P < 0.001) respectively. A negative correlation was observed between the lanreotide levels and GH and IGF-I concentrations in the two groups of patients, but the inhibition of GH/IGF-I concentrations by lanreotide levels was higher in group 1 patients than in those of group 2. Six patients of group 1 were treated with 30mg SR lanreotide injected at monthly intervals. During monthly follow-up, mean GH levels increased above 2.5 microgram/l in 2 patients. After 12 months follow-up, mean GH and IGF-I levels from 4 other patients were similar to those obtained with previous therapeutic sequence (i.e. intramuscular injections every 14 days). CONCLUSION: The degree of responsiveness to lanreotide and the duration of somatotroph suppression following lanreotide withdrawal are variable in acromegalic patients controlled during long-term 30mg SR lanreotide treatment. In patients displaying high sensitivity to lanreotide, the interval between i.m. 30mg SR lanreotide injections can be increased to one month, thus reducing the cost of the therapy, without altering its efficacy upon GH/IGF-I control. PMID- 10822219 TI - Somatic trinucleotide change encompassing codons 882 and 883 of the RET proto oncogene in a patient with sporadic medullary thyroid carcinoma. AB - OBJECTIVE: Restriction analysis is a straightforward procedure for mutational analysis. It is commonly used for screening RET mutations. Incomplete digestion is a well-known cause of false results. Herein, we report another limitation of the method. DESIGN AND METHODS: Screening for somatic mutations in RET exons 16, 13 and 15 was performed in a patient with a sporadic medullary thyroid carcinoma. Genetic study was carried out by both restriction analysis and direct sequencing. RESULTS: A somatic trinucleotide change encompassing codons 882 and 883 of the RET proto-oncogene (GTA GCT to GTT TTT) was documented. Particular to this case is the silent mutation (GTA-->GTT) at codon 882. Independently, both the novel silent mutation and the missense mutation at codon 883 may disrupt the same AluI restriction site. Based on the restriction pattern we were able to say that both mutations occurred in the same allele. CONCLUSIONS: Restriction analysis is an easy approach for screening RET mutations; however, it is not enough to assign a final diagnosis. PMID- 10822220 TI - Relationship between autoantibodies against glutamic acid decarboxylase, thyroglobulin/thyroid microsome and DNA topoisomerase II in the clinical manifestation of patients with type 1 diabetes mellitus in Taiwan. AB - OBJECTIVE: In a preliminary cross-sectional study, we discovered that DNA topoisomerase II autoantibodies (anti-TopII) were detected in 49.2% of 195 Chinese type 1 diabetes mellitus (type 1 DM) patients with a mean age of 14.5 years and a mean duration of disease of 4.6 years. In order to demonstrate the relationship between anti-TopII and other immunological characteristics in Chinese type 1 DM patients, and to evaluate its putative prediction efficacy in Chinese patients, we simultaneously examined the frequency of anti-glutamic acid decarboxylase autoantibodies (anti-GAD), anti-TopII, antithyroglobulin/antimicrosomal autoantibodies (ATA/AMiA) and C-peptide concentrations in our patients in the present study. DESIGN AND METHODS: The frequency of anti-GAD and C-peptide levels, anti-TopII, and ATA/AMiA were examined in our patients by radioimmunoassay, enzyme-linked immunosorbant assay and hemagglutination respectively. Univariate comparisons were performed using Student's t-test for normal distributed data and Chi-square test for diclomatous data. Multivariate analysis was used for interpreting the independent risk factors which increased the incidence of anti-TopII. RESULTS AND CONCLUSIONS: The positivities for anti-GAD, anti-TopII, ATA/AMiA and C-peptide were 45.8%, 50.2%, 13.4% and 11.4% respectively. Anti-GAD and anti-TopII frequencies in our patients were similar when we stratified the patients by age, age at onset and duration. These observations imply that anti-GAD and anti-TopII remain persistent in Chinese patients with long-term type 1 DM duration. The most interesting finding is that anti-TopII frequency is more persistent than anti-GAD in our patients, especially when the diabetic duration is longer than 11 years. This indicates that anti-TopII, rather than anti-GAD, might act as a better indicator for monitoring the pathogenesis of Chinese type 1 DM patients especially in patients with a long-standing duration of disease. The late age of onset (>18 years) is a risk factor which increased the incidence of anti-TopII according to multivariate analysis. We further analyzed different manifestations between the youth- and adult-onset type 1 DM and found that adult-onset type 1 DM is characterized by better preservation of residual beta-cell function and higher frequencies of autoantibodies. PMID- 10822221 TI - Prevalence of positive anti-neutrophil cytoplasmic antibody (ANCA) in patients receiving anti-thyroid medication. AB - OBJECTIVE: Vasculitis is a rare complication of anti-thyroid medications. There are 32 cases of anti-neutrophil cytoplasmic antibody (ANCA)-positive vasculitis in association with anti-thyroid medication reported in the English literature. The objectives of this study were to assess the frequency of positive ANCA in patients on long-term anti-thyroid medication, and to follow patients prospectively from commencement of medication to determine whether they became ANCA-positive after therapy. DESIGN: Prospectively collected cross-sectional study of two groups of patients: (i) who had received long-term (>18 months) anti thyroid medication, and (ii) newly diagnosed thyrotoxicosis before commencement of anti-thyroid medication attending clinic between 28 April 1998 and 30 September 1998. Data were collected for age, sex, ethnicity, underlying thyroid disease, medication and duration, and symptomatology. RESULTS: Eight of 30 patients on long-term anti-thyroid medication (26.7%) were ANCA-positive. All ANCA-positive patients were female, seven were taking propylthiouracil (PTU) at the time of testing. ANCA-positive patients had taken PTU for a mean +/- s.d. of 7.9+/-10.2 years, compared with 0.8+/-2.2 years in ANCA-negative patients (Mann Whitney, P<0.0001). The ten patients with newly diagnosed thyrotoxicosis were ANCA-negative before commencement of carbimazole. One (10%) became ANCA-positive within 8 months of therapy. CONCLUSIONS: In our population, ANCA-positivity in association with long-term anti-thyroid medication is common (26.7%). One patient who was ANCA-negative prior to anti-thyroid therapy has become ANCA-positive. ANCA should be tested in patients receiving long-term anti-thyroid medications, and in patients with adverse reactions. As PTU is more commonly associated with vasculitis and positive ANCA, carbimazole may be the preferred medication for long-term use. Patients with positive ANCA should be followed, and considered for definitive anti-thyroid therapy, to allow cessation of medication. ANCA positivity may resolve after cessation of anti-thyroid medication. PMID- 10822222 TI - Thyroid volumes in a national sample of iodine-sufficient swiss school children: comparison with the World Health Organization/International Council for the control of iodine deficiency disorders normative thyroid volume criteria. AB - OBJECTIVE: The determination of goiter prevalence in children by thyroid ultrasound is an important tool for assessing iodine deficiency disorders. The current World Health Organization/International Council for the Control of Iodine Deficiency Disorders (WHO/ICCIDD) normative values, based on thyroid volume in iodine-sufficient European children, have recently been questioned, as thyroid volumes in iodine-sufficient children from the USA and Malaysia are smaller than the WHO/ICCIDD reference data. Our objective was to describe ultrasonographic thyroid volumes in a representative national sample of iodine-sufficient Swiss school children, and to compare these with the current reference data for thyroid volume. DESIGN AND METHODS: A 3-stage, probability proportionate-to-size cluster sampling method was used to obtain a representative national sample of 600 Swiss children aged 6-12 years. The following data were collected: thyroid size by ultrasound, urinary iodine concentration, weight, height, sex and age. RESULTS: The median urinary iodine concentration (range) of the children was 115 microgram/l (5-413). Application of the WHO/ICCIDD thyroid volume references to the Swiss children resulted in a prevalence of 0%, using either age/sex-specific or body surface area (BSA)/sex-specific cut-off values. Upper limits of normal (97th percentile) of thyroid volume from Swiss children calculated using BSA, sex and age were similar to those reported in iodine-sufficient children in the USA, but were 20-56% lower than the corresponding WHO/ICCIDD references. CONCLUSIONS: Swiss children had smaller thyroids than the European children on which the WHO/ICCIDD references are based, perhaps due to a residual effect of a recent past history of iodine deficiency in many European regions. However, there were sharp differences between our data and a recent set of thyroid volume data in Swiss children produced by the operator and equipment that generated the WHO/ICCIDD reference data. This suggests that interobserver and/or interequipment variability may contribute to the current disagreement on normative values for thyroid size by ultrasound in iodine-sufficient children. PMID- 10822223 TI - From guidelines to hospital practice: reducing inappropriate ordering of thyroid hormone and antibody tests. AB - OBJECTIVE: Because of major technical improvements and conscious care about cost effectiveness, limiting the inadequate use of thyroid biological tests appears to be a major issue. DESIGN: To (i) estimate the ordering prevalence of each thyroid test, (ii) assess the prevalence of relevant thyroid tests, and (iii) evaluate the impact of expressing justification for tests during a 2-month intervention period on these prevalences. METHODS: During a prospective 2-month survey (June July 1997), all the request forms were divided into four groups of prescription: (1) investigation of thyroid function, (2) taking drugs affecting the thyroid, (3) monitoring of nodule and cancer, and (4) investigation of thyroid autoimmunity. Their appropriateness was thus determined according to consensus in our hospital and previously published recommendations. Results were compared with those of retrospective similar 2-month periods in 1996 and 1998. Combinations of thyroid function tests and thyroid antibodies were analyzed during the 1996, 1997 and 1998 periods. RESULTS: The overall estimated rate of appropriate ordering between 1996 and 1997 increased from 42.5% to 72.4% (P<10(-4)), with a significant improvement in each group of main diagnosis referral, except in group 3 where suitability was always over 85%. However, in group 4, appropriateness remained low (36%). Combinations of thyroid tests revealed an increase in single TSH order forms and single autoantibodies to thyroperoxidase (TPOAb) ones, while TSH+free thyroxine+free tri-iodothyronine and TPOAb+ autoantibodies to thyroglobulin ones decreased significantly. Interestingly, all these changes were maintained 1 year later (June-July 1998) even though physicians were not aware of this new study. CONCLUSIONS: Persistent change in medical practice was thus assessed. PMID- 10822224 TI - The value of dehydroepiandrosterone sulfate measurement in the differentiation between benign and malignant adrenal masses. AB - OBJECTIVE: Owing to their increasing rate of discovery as incidental findings, the characterization of adrenal masses is an important diagnostic problem which frequently challenges the clinician's skill. DESIGN: The results of dehydroepiandrosterone sulfate (DHEAS) measurement were evaluated in a consecutive series of 107 patients with an adrenal mass (39 men, 68 women aged 15 81 years, median 56 years). DHEAS levels observed in the patients were categorized as reduced, normal or elevated according to sex- and age-adjusted reference ranges obtained by measuring DHEAS in 214 healthy women and 142 healthy men aged 17-93 years. The working hypothesis was that a low DHEAS level is a marker of benignity. METHODS: In 84 patients, the adrenal mass was discovered serendipitously, while in the remainder the mass was clinically symptomatic. Patients with known extra-adrenal malignancies were excluded. The adrenal masses were categorized as benign or malignant by computerized tomography (CT) criteria. All patients with suspected malignant tumors or with overtly hypersecreting tumors underwent adrenalectomy. The patients with a presumptive benign tumor were followed-up for at least 12 months. RESULTS: In the overall series, the sensitivity and specificity of a low DHEAS level in the identification of a benign lesion were 41% and 100% respectively. Superimposable figures were obtained when considering only adrenal incidentalomas. DHEAS levels in adrenal cancers displayed some overlap with adrenal adenomas, but in only 2/11 adrenal cancers were DHEAS levels normal, while they were elevated in the remainder. CONCLUSIONS: The present data suggest that DHEAS measurement may help to differentiate benign from malignant adrenal masses as a complementary test of CT in a clinical research setting. The value of DHEAS measurement in general practice is limited because it may be difficult to differentiate between low and low-normal values, particularly in the elderly. PMID- 10822225 TI - Use of a long-acting gonadotrophin-releasing hormone analogue in a postmenopausal woman with hyperandrogenism due to a hilus cell tumour. AB - OBJECTIVE: The aim of this study was to prove the utility of GnRH analogues for the suppression of androgen secretion in a postmenopausal woman with a suspected virilizing ovarian tumour. DESIGN AND METHODS: We present a case of a 72-year-old woman with virilization of recent onset. Hormonal studies revealed a fourfold increase in serum testosterone levels, normal dehydroepiandrosterone sulphate concentrations and high levels of serum 17-hydroxyprogesterone levels. Computed axial tomography scan of the ovaries was normal and the adrenal glands showed a discrete enlargement. The long-acting GnRH analogue, triptorelin, was injected initially (3.75mg i.m.) and serum hormone levels were measured weekly throughout one month. RESULTS: GnRH produced a decrease in serum testosterone levels to normal values, in parallel with the suppression of serum LH and FSH concentrations. The patient was treated for three months with triptorelin and she experienced an amelioration of the hyperandrogenic symptoms. In order to achieve a diagnosis, the patient was submitted to a laparotomy that revealed a small hilus cell tumour in the left ovary. CONCLUSION: GnRH analogues may offer a good therapeutic option in some states of gonadotrophin-dependent hyperandrogenism of ovarian origin. PMID- 10822226 TI - Fetal leptin and insulin levels only correlate inlarge-for-gestational age infants. AB - OBJECTIVE: To determine whether fetal leptin levels correlate with fetal weight and whether such correlation is direct or indirect via insulin or human placental lactogen (hPL), respectively. DESIGN: Cross-sectional study of offspring at term (n=175) with over-representation of large-for-gestational age (LGA; n=70) and small-for-gestational age (SGA; n=23) cases in a population of Caucasian women with no pregnancy pathology. METHODS: Fetal cord blood was collected after delivery. In several cases (n=62) paired mother-fetus blood samples were obtained. Leptin, insulin and hPL levels were measured by RIA. Anthropometric data (birth weight, body mass index, placental weight) were recorded. RESULTS AND CONCLUSIONS: Maternal insulin, hPL and leptin levels were higher than fetal concentrations. Cord blood leptin levels positively correlated with the anthropometric data with stronger correlations in female (0.54250 kDa. Expression of the EEM-1 antigen is largely confined to extra-embryonic membranes, but some expression does occur in the embryo. In the embryonic day 6 (E6) and E7 embryo it is expressed in visceral and parietal endoderm; later derivatives of these structures in the yolk sac are negative. The outer layer of the amnion also stains. Within the embryo proper, antigen is expressed in limited regions of the gut, kidney, and pancreas. EEM-1 is also expressed in embryonic stem (ES) cells differentiating in vitro. Undifferentiated ES cells do not express the antigen. Embryoid bodies (EBs) derived from ES cells have patches of EEM-1-positive cells on their surface at 2 days in culture. Older EBs have increasing numbers of positive cells which are confined to the surface. A special class of EBs, termed "cystic EBs," are covered by a cell layer which strongly express EEM-1 antigen. The EEM-1 antibody will be useful for investigating the development of extra embryonic membranes and their counterparts in the ES cell in vitro model. PMID- 10822274 TI - Calphostin C-mediated translocation and integration of Bax into mitochondria induces cytochrome c release before mitochondrial dysfunction. AB - Calphostin C-mediated apoptosis in glioma cells was reported previously to be associated with down-regulation of Bcl-2 and Bcl-xL. In this study, we report that 100 nM calphostin C also induces translocation and integration of monomeric Bax into mitochondrial membrane, followed by cytochrome c release into cytosol and subsequent decrease of mitochondrial inner membrane potential (DeltaPsim) before activation of caspase-3. The integration of monomeric Bax was associated with acquirement of alkali-resistance. The translocated monomeric Bax was partly homodimerized after cytochrome c release and decrease of DeltaPsim. The translocation and homodimerization of Bax, cytochrome c release, and decrease of DeltaPsim were not blocked by 100 microM z-VAD.fmk, a pan-caspase inhibitor, but the homodimerization of Bax and decrease of DeltaPsim were inhibited by 10 microM oligomycin, a mitochondrial F0F1-ATPase inhibitor. Therefore, it would be assumed that mitochondrial release of cytochrome c results from translocation and integration of Bax and is independent of permeability transition of mitochondria and caspase activation, representing a critical step in calphostin C-induced cell death. PMID- 10822275 TI - The staurosporine analog, Ro-31-8220, induces apoptosis independently of its ability to inhibit protein kinase C. AB - A series of bisindolylmaleimide (Bis) compounds were designed as analogs of the natural compound staurosporine (STS), which is a potent inducer of apoptosis. Many of the Bis analogs appear to be highly selective inhibitors of the protein kinase C (PKC) family, including PKC-alpha, -beta, -gamma, -delta, -epsilon, and zeta, unlike STS, which is an inhibitor of a broad spectrum of protein kinases. In this report we describe the effects of the Bis analogs, Bis-I, Bis-II, Bis-III and Ro-31-8220 on the survival and proliferation of HL-60 cells, which have been widely used as a model cell system for studying the biological roles of PKC. Treatment of HL-60 cells with Bis-I, Bis-II, Bis-III, or Ro-31-8220 blocked phosphorylation of the PKC target protein Raf-1 with equal potency but did not appear to affect the general phosphorylation of proteins by other kinases. However, the biological effects of the Bis compounds were different: Bis-I and Bis-II had no observable effects on either cell survival or proliferation; Bis III inhibited cell proliferation but not survival, whereas Ro-31-8220 induced apoptosis. These results indicated that the members of the PKC family which could be inhibited by the Bis analogs were required neither for survival nor proliferation of HL-60 cells. Analyses of cells treated with Ro-31-8220 showed that the apoptotic effect of Ro-31-8220 on HL-60 cells was mediated by a well characterized transduction process of apoptotic signals: i.e., mitochondrial cytochrome c efflux and the activation of caspase-3 in the cytosol. Moreover, the ability of Ro-31-8220 to induce apoptotic activation was completely inhibited by the over-expression of the apoptotic suppressor gene, Bcl-2, in the cells. Interestingly, proliferation of the Bcl-2-over-expressing cells was still sensitive to the presence of Ro-31-8220, suggesting that the inhibitory effects of Ro-31-8220 on viability and cell proliferation were mediated by different mechanisms. In particular, the apoptotic effect of Ro-31-8220 on cells was not altered by the presence of an excess amount of the other Bis analogs, suggesting that this effect is mediated by a factor(s) other than PKC or by a mechanism which was not saturable by the other Bis analogs. Finally, structure-function analyses of compounds related to Ro-31-8220 revealed that a thioamidine prosthetic group in Ro-311-8220 was largely responsible for its apoptotic activity. PMID- 10822276 TI - Increase in ultraviolet sensitivity by overexpression of calpastatin in ultraviolet-resistant UVr-1 cells derived from ultraviolet-sensitive human RSa cells. AB - Human RSa cells are highly sensitive to apoptotic-like cell death by ultraviolet irradiation (UV) while UVr-1 cells are their variant with an increased resistance to UV. Three days after UV at 10 J/m2, the viability of RSa cells was approximately 17% while that of UVr-1 cells was 65%. This different survival might reflect apoptotic cell death since apoptosis-specific DNA ladder was more clearly observed in RSa cells than in UVr-1 cells after UV. Addition of ALLN/calpain inhibitor I to the culture medium after UV resulted in similar survival (14 - 18%) between RSa and UVr-1 cells. Immunoblot analysis showed down regulation of protein kinase CTheta, Src, Bax and mu-calpain after UV was more prominent in UVr-1 than in RSa cells. Activated mu-calpain appeared within 1 h post-UV only in UVr-1 cells. The expression of calpastatin, a specific endogenous inhibitor of calpain, was higher in RSa than in UVr-1 cells. To further examine the role of calpain in UV-induced cell death, cDNA of human calpastatin was transfected into UVr-1 cells. The results showed that overexpression of calpastatin suppressed down-regulation of Src, mu-calpain and Bax. Concomitantly, colony survival after UV was reduced in calpastatin-transfected cells as compared to vector control cells. Our results suggest that activation of calpain might account for, at least in part, the lower susceptibility to UV-induced cell death in UVr-1 cells. PMID- 10822277 TI - CD 95-independent mechanisms of IL-2 deprivation-induced apoptosis in activated human lymphocytes. AB - Growth factor deprivation-induced apoptosis plays an important role in several cellular systems. However, knowledge of the molecular mechanisms involved are restricted to a few murine models or tumor cell lines. Therefore, we aimed studying signaling pathways leading to apoptosis in activated human peripheral T cells after IL-2 withdrawal. Lymphoblasts from patients with CD 95 (Fas/APO-1) deficiency revealed that functional CD95 was not required to induce apoptosis after IL-2 withdrawal. Moreover, apoptosis induction in response to various cytotoxic stimuli was found to be mediated in the absence of functional CD95 but was affirmatorily influenced by IL-2 signaling. Immunoblots showed no downregulation of Bcl-2 or Bcl-xL and no upregulation of Bax, whereas decreased mitochondrial membrane potential was readily measurable 24 h after cytokine deprivation. Tetrapeptide inhibitors showed limited efficacy in preventing apoptosis whereas the caspase inhibitor zVAD-FMK potently blocked induction of apoptosis. Cleavage of different fluorogenic substrates revealed multiple caspase enzyme activities in lymphoblasts, which were not negatively affected by the fas mutation. Starting at 8 h after IL-2 withdrawal, upregulation of active caspase-3 but not of caspase-8 could be detected. Taken together, our data argue for molecular mechanisms of cytokine deprivation-induced apoptosis in activated human lymphocytes independent of CD95. PMID- 10822278 TI - Morphological and molecular evidence of differentiation during etoposide-induced apoptosis in human lymphoblastoid cells. AB - The relationship between apoptosis and cell differentiation has been a subject for continuous debate, with evidence showing leukaemic cell differentiation and drug-induced apoptosis have reciprocal, interdependent and a highly schedule dependent relationship. We have addressed this relationship in terms of a widely used model for apoptosis induced by cytotoxic drugs: namely the effect of etoposide on CEM cells. In respect of commitment toward differentiation, we assessed changes in expression of marker genes and the level of CD3 antigenicity. Changes in these parameters following exposure of CEM cells to etoposide was similar to that observed following treatment of the same cells with phorbol 12 myristate 13-acetate (PMA), though this latter treatment did not cause cell death. Similarities in response to etoposide and PMA also included generation of 50 kilobase fragmentation of DNA and convolution of nuclei as assessed by transmission electron microscopy. However, condensation of chromatin and externalization of phosphatidylserine were only recorded in response to the cytotoxic drug and not in response to PMA. The data are consistent with apoptosis in these lymphoblastoid cells being accompanied by activation of specific markers of T-cell differentiation, but ultimately involving processes unequivocally associated with cell death. PMID- 10822279 TI - Cleavage of BID during cytotoxic drug and UV radiation-induced apoptosis occurs downstream of the point of Bcl-2 action and is catalysed by caspase-3: a potential feedback loop for amplification of apoptosis-associated mitochondrial cytochrome c release. AB - BID, a pro-apoptotic Bcl-2 family member, promotes cytochrome c release during apoptosis initiated by CD95L or TNF. Activation of caspase-8 in the latter pathways results in cleavage of BID, translocation of activated BID to mitochondria, followed by redistribution of cytochrome c to the cytosol. However, it is unclear whether BID participates in cytochrome c release in other (non death receptor) cell death pathways. Here, we show that BID is cleaved in response to multiple death-inducing stimuli (staurosporine, UV radiation, cycloheximide, etoposide). However BID cleavage in these contexts was blocked by Bcl-2, suggesting that proteolysis of BID occurred distal to cytochrome c release. Furthermore, addition of cytochrome c to Jurkat post-nuclear extracts triggered breakdown of BID at Asp-59 which was catalysed by caspase-3 rather than caspase-8. We provide evidence that caspase-3 catalysed cleavage of BID represents a feedback loop for the amplification of mitochondrial cytochrome c release during cytotoxic drug and UV radiation-induced apoptosis. PMID- 10822280 TI - Inducible expression of filaggrin increases keratinocyte susceptibility to apoptotic cell death. AB - Filaggrin is an intermediate filament associated protein that aids the packing of keratin filaments during terminal differentiation of keratinocytes. Premature aggregation of keratin filaments is prevented by filaggrin expression as the inactive precursor, profilaggrin, which is localized in keratohyalin granules in vivo. We have previously shown that filaggrin constructs, when transiently transfected into epithelial cells, lead to a collapsed keratin cytoskeletal network and dysmorphic nuclei with features of apoptosis. The apparent transfection rate is low with filaggrin constructs, supporting their disruptive role but hindering further study. To bypass this problem, we generated stable keratinocyte cell lines that express mature human filaggrin using a tetracycline inducible promoter system. We found that cell lines expressing filaggrin, but not control cell lines, exhibited increased sensitivity to multiple apoptotic stimuli as measured by morphologic and biochemical criteria. None of the cell lines showed an increase in endogenous expression of filaggrin in response to the same stimuli. Filaggrin expression alone was insufficient to induce apoptosis in these keratinocyte cell lines. We conclude that filaggrin, due to its keratin binding ability, primes cells for apoptosis. Because filaggrin is expressed at a level of the epidermis where keratinocytes are in transition between the nucleated granular and the anucleate cornified layers, we hypothesize that filaggrin aids in the terminal differentiation process by facilitating apoptotic machinery. PMID- 10822281 TI - Susceptibility to drug-induced apoptosis correlates with differential modulation of Bad, Bcl-2 and Bcl-xL protein levels. AB - To define the responses of apoptotic regulatory proteins to different chemotherapeutic agents, we investigated the expression of Bcl-2 family gene products, the release of cytochrome c, and the activation of pro-caspase-3 during apoptosis induced by Taxol and Thiotepa, in the MCF-7 breast carcinoma and the HL 60 leukemia cell lines. The earliest event induced by drug exposure was increase in Bad protein levels, followed by Bcl-2 down-regulation, cytochrome c release, and Bcl-xL and Bax up-regulation. Bak accumulation was a late event. Activation of pro-caspase-3 and cleavage of Bcl-2 protein occurred in the HL-60 cells only, and followed the cytochrome c release. The overall responses were qualitatively similar in both cell types, but MCF-7 cells treated with Taxol showed a significant delay in apoptosis, correlating with early up-regulation of Bcl-2 and delayed release of cytochrome c. We conclude that Bad up-regulation is an early indicator of a cellular response that will lead to cell death, but may be modulated by survival mechanisms, which cumulatively govern the ultimate susceptibility to apoptosis. PMID- 10822282 TI - Reliability of fatty acid composition in human serum phospholipids. AB - OBJECTIVE: We examined the reliability of the fatty acid composition of serum phospholipids in the New York University Women's Health Study, a prospective study of sex hormones, diet and breast cancer. DESIGN: Non-fasting serum samples collected at three yearly visits, in 46 healthy women, and stored at -80 degrees C for 7-12 y, were included in the study. Serum phospholipid fatty acid composition was measured by capillary gas chromatography. RESULTS: For the 20 individual fatty acids measured, the reliability coefficients were less than 0.50 for four, between 0.50 and 0.70 for nine, and greater than 0.70 for seven. Among the major fatty acids, arachidonic and alpha-linolenic acids had high reliability coefficients (0.71 and 0. 72, respectively), palmitic, oleic, linoleic, eicosapentaenoic and docosahexaenoic acids had intermediate coefficients (0.57, 0.69, 0. 62, 0.64 and 0.66, respectively), whereas stearic acid had the lowest coefficient (0.15). The reliability coefficients for total monounsaturated fats, omega-6 and omega-3 fatty acids were moderately high (0.66, 0.53 and 0.66, respectively), whereas the coefficients for total saturated fats and total polyunsaturated fats were low (0.31 and 0.43, respectively). CONCLUSIONS: These results indicate that the fatty acid composition of serum phospholipids can be a useful tool in epidemiologic studies, although for most fatty acids a single determination is associated with some error in measurement that should be taken into account at the design and analysis stages. Storage for up to 12 y at -80 degrees C preserved polyunsaturated fatty acids from oxidation very well. PMID- 10822283 TI - Extruded dry beans and serum lipoprotein and plasma haemostatic factors in hyperlipidaemic men. AB - OBJECTIVE: To examine the effects of the inclusion of extruded dry beans in the diet on serum lipoprotein, plasma fibrinogen, plasma viscosity and plasminogen activator inhibitor 1 (PAI-1) levels. SUBJECTS AND STUDY DESIGN: Twenty-two free living hyperlipidaemic men participated in this randomised, controlled, cross over study. The subjects were randomly assigned to one of two groups. After a run in period of four weeks, during which subjects followed their normal diet with the exclusion of dry beans, group A had to include 110 g/day of extruded dry beans in the form of baked products for four weeks while group B continued with the run-in diet. A washout period of four weeks followed after which the experimental intervention was crossed-over. Anthropometric measurements, serum lipoproteins and haemostatic variables were measured with standard methods and dietary intakes were estimated with five-day dietary records at the beginning and end of each experimental period. RESULTS: Compliance was determined as 83.5% with a mean intake of 91. 9 g/day extruded dry beans. Extruded dry beans did not have significant effects on total serum cholesterol, low density lipoprotein cholesterol, triglycerides, apolipoprotein A or B, plasma fibrinogen and plasma viscosity concentrations. High density lipoprotein cholesterol concentrations decreased in both the dry bean and control periods. Lipoprotein (a) concentrations increased with intake of extruded dry beans, but this increase was probably not due to an independent effect of extruded dry beans. Plasminogen activator inhibitor 1 levels were significantly lower after the intake of extruded dry beans compared to the control period. CONCLUSIONS: The inclusion of 91.9 g extruded dry beans per day in the diet had no effects on serum lipoproteins, plasma fibrinogen and viscosity levels but decreased PAI-1 levels. SPONSORSHIP: Dry Bean Producers Organisation (South Africa) and the Potchefstroom University for Christian Higher Education, Potchefstroom, South Africa. PMID- 10822284 TI - Glycaemic index of parboiled rice depends on the severity of processing: study in type 2 diabetic subjects. AB - OBJECTIVE: To study the influence of parboiling and the severity of the process on glycaemic and insulinaemic responses to rice in type 2 diabetes. Moreover, to examine changes in starch structure related to parboiling, which may affect the metabolic responses and digestibility. DESIGN: Nine type 2 diabetic subjects ingested four test meals: white bread (WB) and three meals of cooked polished rice of the same variety being non-parboiled (NP), mildly traditionally parboiled (TP) and severely pressure parboiled (PP). The participants ingested the test meals (50 g available carbohydrates) on separate occasions after an overnight fast. SETTING: Outpatient clinic, Dept. Endocrinology and Metabolism, Aarhus University Hospital, Denmark. RESULTS: All three rice samples elicited lower postprandial plasma glucose response (NP: 335+/-43; TP: 274+/-53; PP: 231+/-37 mmol/1*180 min.; means+/-s.e.m.) than white bread (626+/-80; P<0.001), within rice samples PP tended to be lower than NP (P=0.07). The glycaemic indices were: NP: 55+/-5, TP: 46+/-8 and PP: 39+/-6, and lower for PP than NP (P<0.05). The insulin responses were similar for the three rice meals, which were all lower than that to white bread (P<0.001). Differential scanning calorimetry showed the presence of amylose-lipid complexes in all rice samples and of retrograded amylopectin in PP. Amylose retrogradation was not detected in any of the rice samples. CONCLUSIONS: All rice test meals were low-glycaemic in type 2 diabetic subjects. There was no effect of TP on glycaemic index, whereas PP reduced the glycaemic index by almost 30% compared to NP. SPONSORSHIP: The Royal Veterinary and Agricultural University, Aarhus University Hospital, Danish International Development Assistance (DANIDA), Ministry of Foreign Affairs and the 'Konsul Johannes Fogh-Nielsens og Fru Ella Fogh-Nielsens Legat' foundation. PMID- 10822285 TI - Within- and between-subject variation in energy expenditure measured by the doubly-labelled water technique: implications for validating reported dietary energy intake. AB - OBJECTIVES: To estimate the total (CVt), within-subject (CVw) and between-subject (CVb) variation in free-living energy expenditure as measured by the doubly labelled water (DLW) technique. To examine the limitation of the DLW measurement of energy expenditure for evaluating reported energy intake. To estimate the probable minimum and maximum 'habitual' energy expenditures for a sustainable lifestyle. DESIGN: Review and analysis of individual data from 25 studies with repeat DLW measurements of energy expenditure (EE). RESULTS: Pooled mean CVw derived from 21 studies was 11.8% for EE and 12.3% for physical activity level (PAL). Multiple regression analysis of CVw in 25 studies found a positive association with time span between measurements. At zero time CVw for EE was 8.2% rising to 9.6% at 13 weeks and 15.4% at 52 weeks. At the same time points CVw for PAL was 9.1%, 10.0% and 13.4% respectively. Pooled mean CVt was 13.0% for EE and 10.7% for PAL. CVb calculated from pooled mean CVt and CVw was 20.6% for EE and 7.2% for PAL. 95% confidence limits of PAL in 11 age-sex groups averaged 1.2 to 2.2. CONCLUSIONS: The analysis supported previous estimates of 8% for within subject variation in DLW measurements including analytic plus inherent biologic variation. Variation that included changes in weight, season and activity increased with increased time between measurements to about 15% at a time span of 12 months. Confidence limits of agreement between EE and reported energy intake were estimated to range from +/-15% to +/-32%. Estimates of the range of usual EE for normally active persons ranged from 1.3 to 2.2. PMID- 10822286 TI - The sensitivity and specificity of the Goldberg cut-off for EI:BMR for identifying diet reports of poor validity. AB - OBJECTIVE: To explore the specificity and sensitivity of the Goldberg cut-off for EI:BMR for identifying diet reports of poor validity as compared with the direct comparison of energy intake with energy expenditure measured by doubly-labelled water. DESIGN: Twenty-two studies with measurements of total energy expenditure by doubly-labelled water (EE), basal metabolic rate (BMR) and energy intake (EI) provided the database (n=429). The ratio EI:EE provided the baseline definition of under- (UR), acceptable- (AR) and over-reporters (OR), respectively EI:EE <0.76, 0.76-1.24 and >1.24. Four strategies for identifying under- and over reporters using the Goldberg cut-off were explored. Sensitivity of the cut-off was calculated as the proportion of UR correctly identified and specificity as the proportion of non-UR correctly identified. RESULTS: UR, AR and OR (by EI:EE) were 34, 62 and 4% respectively of all subjects. When a single Goldberg cut-off for the physical activity level (PAL) of 1.55 was used, for men and women respectively the sensitivity was 0.50 and 0.52 and the specificity 1. 00 and 0.99. Using a cut-off for higher PAL traded specificity for sensitivity. Using the cut-off for a PAL of 1.95, sensitivity was 0. 76 and 0.85 and the specificity 0.87 and 0.78 for men and women respectively. Using cut-offs for mean age-sex specific PAL did not improve sensitivity. When subjects were assigned to low, medium and high activity levels and cut-offs for three different PALs used, sensitivity improved to 0.74 and 0.67 without loss of specificity (0. 97 and 0.98), for men and women respectively. If activity levels for men were applied to the womens' data, sensitivity improved to 0.72. CONCLUSION: To identify diet reports of poor validity using the Goldberg cut-off for EI:BMR, information is needed on each subject's activity level. PMID- 10822287 TI - Effects of blueberry and cranberry juice consumption on the plasma antioxidant capacity of healthy female volunteers. AB - OBJECTIVE: To assess whether consumption of 500 ml of blueberry juice or cranberry juice by healthy female subjects increased plasma phenolic content and antioxidant capacity. DESIGN: Latin square arrangement to eliminate ordering effects. After an overnight fast, nine volunteers consumed 500 ml of blueberry juice, cranberry juice or a sucrose solution (control); each volunteer participated on three occasions one week apart, consuming one of the beverages each time. Blood samples were obtained by venipuncture at intervals up to four hours after consumption of the juices. Urine samples were also obtained four hours after consuming the juice. RESULTS: Consumption of cranberry juice resulted in a significant increase in the ability of plasma to reduce potassium nitrosodisulphonate and Fe(III)-2,4, 6-Tri(2-pyridyl)-s-triazine, these measures of antioxidant capacity attaining a maximum after 60-120 min. This corresponded to a 30% increase in vitamin C and a small but significant increase in total phenols in plasma. Consumption of blueberry juice had no such effects. CONCLUSION: The increase in plasma antioxidant capacity following consumption of cranberry juice could mainly be accounted for by an increase in vitamin C rather than phenolics. This also accounted for the lack of an effect of the phenolic rich but vitamin C-low blueberry juice. SPONSORSHIP: Funded by the Scottish Executive Rural Affairs Department and the Danish Government. PMID- 10822288 TI - A combined heart rate and movement sensor: proof of concept and preliminary testing study. AB - OBJECTIVE: Heart rate monitoring has previously been used as a technique for measuring energy expenditure (EE) in field studies. However, the combination of heart rate monitoring with movement sensoring could have theoretical advantages compared to either method used alone. Therefore, this study was undertaken to develop and validate a new combined heart rate monitor and movement sensor instrument (HR+M) for measuring EE. METHODS: The HR+M instrument is a single piece instrument worn around the chest which records minute-by-minute heart rate and movement. Eight subjects underwent an individual calibration in which EE and heart rate were measured at rest and during a sub-maximal bicycle ergometer test. They then wore the HR+M for 24 hours in a whole-body calorimeter and underwent a standard protocol including periods of physical activity and inactivity. Minute by-minute heart rate was converted to EE using individual calibration curves with the motion data discriminating between periods of inactivity and activity at low heart rate levels. EE was also calculated using the HRFlex method which relies on heart rate alone. Both estimates of EE were compared to EE measured in the whole body calorimeter. RESULTS: The mean percentage error of the HR+M method calculating TEE compared with the gold standard of the calorimeter measurement was 0.00% (95% CI of the mean error -0.25, 1. 25). The HRFlex method using the heart rate information alone resulted in a mean percentage error of 16.5% (95% CI of the mean error -0.57, 1.76). CONCLUSIONS: This preliminary test of HR+M demonstrates its ability to estimate EE and the pattern of EE and activity throughout the day. Further validation studies in free-living individuals are necessary. SPONSORSHIP: NJW is an MRC Clinician Scientist Fellow. KLR holds an MRC PhD scholarship. PMID- 10822289 TI - Quercetin intake and the incidence of cerebrovascular disease. AB - OBJECTIVE: To study the relation between intake of the antioxidant flavonoid quercetin and subsequent incidence of cerebrovascular disease (CVA). DESIGN: A cohort study carried out among 9208 Finnish men and women 15 y or more of age and initially free from cardiovascular disease. During a 28 y follow-up period in 1967-1994, a total of 824 cases with CVA were diagnosed. METHODS: Food consumption data were collected using a dietary history interview method covering the total habitual diet during the previous year. RESULTS: Quercetin intake was not associated with CVA incidence. The relative risk of CVA adjusted for age, serum cholesterol, body mass index, smoking, hypertension, diabetes, geographical area, occupation and intake of beta-carotene, vitamin E, vitamin C, fibre, various fatty acids, and energy between the highest and lowest quartiles of quercetin intake was 0.99 (95% confidence interval (CI)=0.71-1.38) for men and 0.85 (CI=0.60-1.21) for women. In contrast, apples, the major source of quercetin in the study population, showed a significant inverse association both in men and women, mainly due to an association with thrombotic or embolic stroke. The relative risks of thrombotic stroke after further adjustment for quercetin intake were 0.59 (CI=0.35-0.99; P=0.45) and 0.61 (CI=0.33-1.12: P for trend=0.02) for men and women, respectively. CONCLUSIONS: The results suggest that the intake of apples is related to a decreased risk of thrombotic stroke. This association apparently is not due to the presence of the antioxidant flavonoid quercetin. PMID- 10822290 TI - Similar responses in hemostatic factors after consumption of wholemeal rye bread and low-fiber wheat bread. AB - OBJECTIVE: Both epidemiological and experimental evidence suggests a protective effect of dietary fiber against disease, such as cancer and cardiovascular disease. The effects of fiber on lipoproteins are modest, indicating that other mechanisms may be involved. As the hemostatic effects of different types of fiber are poorly known, we compared the effects of wholemeal rye bread and low-fiber wheat bread on factors related to coagulation, fibrinolysis and platelet function. DESIGN: Subjects consumed rye and wheat bread as part of their habitual diet in a cross-over manner for 4 weeks, with a 4 week washout period between the diet periods. SUBJECTS: Forty healthy subjects (18 men, 22 women), aged 43+/-2 y. RESULTS: Mean daily intake of fiber during the rye bread period was 31 g for men and 26 g for women, while the respective figures for the wheat bread period were 15 g and 12 g. However, no significant differences between the two periods were seen in factor VII coagulant activity (FVIIc), fibrinogen, prothrombin fragments 1+2 (F1+2), tissue plasminogen activator (tPA) plasminogen activator inhibitor 1 (PAI-1) or 2,3,-dinor-thromboxane B2. Fibrin degradation products, D-dimers, were slightly lower after the wheat period (P=0.046). CONCLUSIONS: The results indicate that wheat and rye bread do not differ in their effects on coagulation, fibrinolysis or platelet function. SPONSORSHIP: Fazer Bakeries Ltd, Lahti Finland; Vaasan & Vaasan Ltd, Helsinki, Finland. PMID- 10822291 TI - Low serum folate concentrations are associated with an excess incidence of acute coronary events: the Kuopio Ischaemic Heart Disease Risk Factor Study. AB - OBJECTIVE: To test the hypothesis that low serum folate concentrations are associated with an increased risk of acute coronary events in men free of prior coronary heart disease. SETTING: Research Institute of Public Health, University of Kuopio, Kuopio, Finland. DESIGN: Prospective study in a cohort of 734 men aged 46-64 y examined in 1991-1993 as part of the Kuopio Ischaemic Heart Disease Risk Factor Study (KIHD) and followed for 5 y and 3 months. INTERVENTION: Acute coronary events during the follow-up period were obtained by national hospital discharge registry. Baseline serum folate concentrations were measured by radioimmunoassay. RESULTS: During the follow-up, six (2.5%) men with higher serum folate concentrations (highest third>11.3 nmol/1) and 28 (5.7%) men with lower serum folate (two lowest thirds) developed an acute coronary event (P=0.008). In a Cox model adjusting for age, examination years, and plasma lycopene concentration, in men with higher serum folate concentrations the relative risk for an acute coronary event was 0.31 (95% CI 0.11-0.90, P=0.031) when compared with men with lower serum folates. CONCLUSION: This prospective cohort study in middle-aged men from eastern Finland indicates that moderate-to-high levels of serum folate are associated with a greatly reduced incidence of acute coronary events. PMID- 10822292 TI - Changes in iodine metabolism during late pregnancy and lactation: a longitudinal study among Sudanese women. AB - OBJECTIVE: To elucidate the persistence, or otherwise, of the pregnancy-related changes in the iodine metabolism and thyroid function in a population residing in an area of mild iodine deficiency in the Sudan. DESIGN: A longitudinal prospective cohort study involving pregnant women who were recruited during their third trimester of pregnancy and were followed up for up to nine months after delivery. SETTING: The study was conducted among Sudanese women residing in the Omdurman area in Khartoum, an area with a total goitre rate of 17.5%. SUBJECTS: Forty-seven pregnant women were recruited during their third trimester of pregnancy. Their mean age and weight were 29+/-4.6 y and 62.4+/-8.7 kg, respectively. Age matched healthy non-pregnant women living in the same area (n=40) served as a control group. METHODS: Serum levels of thyrotropin (TSH), thyroxine (T4), triiodothyronine (T3), thyroglobulin (Tg) and the urinary iodine concentration (UIC) were determined during the third trimester (first occasion), and subsequently at three months (second occasion), six months (third occasion) and nine months after delivery (fourth occasion). Control subjects provided corresponding samples on one occasion. MAIN OUTCOMES MEASURES: UIC, TSH, Tg, FT4 and T3. RESULTS: During the third trimester of pregnancy the median UIC and the free T4 (FT4) were lower than in the control group (P<0. 0001, P<0.0001, respectively), while the median Tg was higher than in the controls (P<0.03). Three months post-partum maximum thyroidal stimulation was evident, with elevated serum levels of TSH and Tg (P<0.0001, P<0.03) and reduced UIC and serum T3 and FT4 (P<0.0004, P<0.0005, P<0.0001), compared with the control group. Nine months post-partum the thyroid function was restored to the pre-pregnancy state, and the median values of TSH, Tg, T3, FT4 and UIC did not differ significantly from those in the control group. CONCLUSIONS: Our study suggests that the reversibility of the pregnancy-induced changes in the iodine status and thyroid function to the pre-pregnancy levels may depend on the iodine status of the mother during and after pregnancy. The thyroidal stress during the first three months of the post partum period, which is partially due to the iodine loss in the breast milk, justifies further detailed studies to assess the iodine content of the breast milk and the role of breast milk as a vehicle for iodine supply of infants in situations of iodine deficiency. SPONSORSHIP: This study was supported by a grant from the Swedish Agency for Research Co-operation with Developing Countries. PMID- 10822293 TI - A cross-sectional study of dietary habits and urinary glucose excretion - a predictor of non-insulin-dependent diabetes mellitus. AB - OBJECTIVE: To investigate the association between consumption of certain foods and macronutrients and urinary glucose excretion, which is a predictor of non insulin-dependent diabetes mellitus. DESIGN: A cross-sectional study, Denmark, 1993-97. SUBJECTS: Participants in the Danish study 'Diet, Cancer and Health'. After exclusion of persons with postprandial urine samples and persons with diabetes or other diseases potentially resulting in glycosuria, the study population included 14 743 men and 18 064 women aged 50-64 y. We identified 183 men and 43 women with glucose in their urine. RESULTS: Consumption of poultry was negatively associated with glycosuria in both men (odds ratio, OR=0.87; 95% confidence interval, 95% CI=0.77-0.98) and women (OR=0.69; 0.48-1.00). Fiber from fruit showed a weak negative association with glycosuria in both men (0. 95; 0.90 1.01) and women (0.89; 0.78-1.02), whereas a significant negative association with total fiber (0.68; 0.51-0.91) and fiber from vegetables (0.94; 0.88-0.99) was seen in men. Intake of fish tended to reduce the risk of glycosuria in women only (0.80; 0.63-1. 02), whereas ingestion of milk products increased their risk significantly (1.15; 1.06-1.24). CONCLUSION: Although statistical significance and consistency in the two sexes were not achieved for all end-points, the study indicates a protective effect of dietary products like poultry, fruit and cereals against glycosuria and suggests a promoting effect of milk. SPONSORSHIP: The Danish National Board of Health and the Danish Cancer Society. PMID- 10822294 TI - Serum electrolytes in hospitalized pellagra alcoholics. AB - BACKGROUND: Owing to high diarrhoea and protein malnutrition frequencies in pellagra, we hypothesised that pellagra patients would have higher electrolyte disturbances than non-pellagra alcoholics. OBJECTIVE: To compare serum electrolytes of hospitalised alcoholics with or without pellagra. DESIGN: Retrospective and descriptive case-control study. SETTING: Internal Medicine wards at a University Hospital, Medical School of Uberaba, Brazil. SUBJECTS: Medical records were reviewed to obtain relevant clinical details, main diagnosis and laboratory data, including serum electrolytes on hospital admission of pellagra patients (n=33) and a randomly chosen control group of alcoholics (n=37), matched in age, gender and socio-economic status. Anaemia was ascertained by haemoglobin <12.5 g/dl (men) and 1.5 g/dl (women), and hypoalbuminemia by serum albumin <3.3 g/dl. RESULTS: Pellagra and controls showed similar age (39.4+/-13.1 vs 45.0+/-11.4 years) and a male predominance of gender (69.7 vs 78.4%), and similar associated diagnoses, including high blood pressure (21.2 vs 16.2%), peripheral neuropathy (12.1 vs 13. 5%), and pneumonia (9.1 vs 13.5%). Despite displaying similar serum sodium (136.6+/-6.1 vs 137.8+/-5.7 mEq/I), magnesium (1.72+/-0.74 vs 1.62+/-0.34 mg/dl), phosphorus (3.79+/-0.87 vs 3.87+/ 0.78 mEq/1) than controls,in addition to higher hypoalbuminemia (76.2 vs 33%) and anaemia (60.6 vs 35.1%) frequencies. CONCLUSIONS: Higher anaemia and hypoalbuminemia frequencies associated with lower serum potassium levels suggest increased protein malnutrition prevalence among pellagrins. PMID- 10822295 TI - Nutritional intakes of vegetarian populations in France. AB - OBJECTIVE: To assess food behaviour and determine nutritional intakes of various vegetarian populations in France. DESIGN: A five-day self-administered food record which was mailed to members of the three principal French vegetarian organisations. SUBJECTS: 145 subjects, aged 7-87 y; 94 classical vegetarians (19% of those contacted), 34 Hindu lactovegetarians (17% of those contacted) and 17 macrobiotic (34% of those contacted). SETTING: The survey was conducted between March 1997 and July 1997 in France. RESULTS: Vegetarianism in France is represented by three main classes of food behaviour: ovolactovegetarian (AAV), lactovegetarian (KRI) and macrobiotic (MMK). The geometric mean intakes ranged from 1952 kcal/d (KRI), 2051 kcal/d (MMK) to 2384 kcal/d (AAV) for males and from 1302 kcal/d (MMK), 1675 kcal/d (AAV) to 1804 kcal/d (KRI) for females, after adjusting for age and BMI. The energy consumption in the MMK group was significantly lower than that in the AAV (P<0.05) and KRI groups (P<0.01), respectively. A difference among groups was observed for females (P=0.0002), but not for males. The MMK group consumed less lipid than the other two vegetarian groups, 46 g/d for men and 38 g/d for women vs 80 g/d for men and 61 g/d for women in the AAV group and 93 g/d for men and 81 g/d for women in the KRI group, respectively. Differences with AAV and MMK were statistically significant (P<0.001 for men and women for both groups). Mean protein consumption ranged from 60 g/d (AAV), 64 g/d (KRI) to 77 g/d (MMK) for males and from 46 g/d (MMK), 50 g/d (AAV) to 58 g/d (KRI) for females. Mean carbohydrate intakes ranged from 247 g/d (AAV), 321 g/d (KRI) to 338 g/d (MMK) in males and from 209 g/d (MMK), 228 g/d (AAV) to 242 g/d (KRI) in females. There were no significant differences in protein and carbohydrate intakes between the groups. Median calcium intakes ranged from 758.2 mg/d (MMK), 863 mg/d (AAV) to 989.3 mg/d (KRI) for the men and from 500.8 mg/d (MMK), 863 mg/d (AAV) to 934 mg/d (KRI) for the women. In the men, there was no differences in daily calcium intakes between the three vegetarian groups. However, we found a significant difference for women (P=0. 0041). The women in the MMK group presented significantly lower daily calcium intakes than the women in the AAV (P=0.013) and KRI (P=0.0032) groups. The AAV and KRI groups consumed dairy products supplying respectively 36% and 53% for the men and 39% and 59% for the women of total calcium against 0% for men and women in the MMK group. Median iron intakes ranged between 12.5 mg/d (KRI), 13.2 mg/d (AAV) and 22.5 mg/d (MMK) for the men and between 11.2 mg/d (KRI), 14.6 mg/d (AAV) and 16.9 mg/d (MMK) for the women. MMK (men P=0.0172 and women P=0.0131) and AAV (only in men P=0.037) groups consumed significantly higher quantities of iron than did the KRI group. The heme iron median intake in males and females of the three vegetarian groups was very low (<0.5%). Overall, the female vegetarians consumed 58.1 (MMK), 109 (AAV) and 127.4 (KRI) mg of vitamin C per day and the males 76.3 (MMK), 150.4 (AAV) and 150.4 (KRI) mg per day. Median vitamin B9 intakes ranged from 247.5 microg/d (KRI), 312 microg/d (MMK) to 390.4 microg/d (AAV) for the men and from 188.3 microg/d (MMK), 266.9 microg/d (KRI) to 323.8 microg/d (AAV) for the women. Vitamin B12 consumption ranged from 0.2 microg/d (MMK), 1.5 microg/d (AAV) to 1.7 microg/d (KRI) in the women and from 0.6 microg/d (MMK) to 1.0 microg/d (AAV and KRI) in the men. No differences in consumption were observed in the males. On the other hand, the women in the MMK group consumed significantly less vitamin C and B12 than did the women in the AAV (P=0.0006) and KRI (P=0. 0396) groups, while it was at the limit of significance for the females (P=0.0715) for vitamin B9. CONCLUSION: Our results suggest that vegetarians have a better understanding of dietary requirements than does the general population. (ABSTRACT TRUNCATED) PMID- 10822296 TI - Effect of peritoneal fluid on whole body and segmental multiple frequency bioelectrical impedance in patients on peritoneal dialysis. AB - OBJECTIVE: We investigated the ability of whole body and segmental multiple frequency bioelectrical impedance (MFBIA) to detect peritoneal fluid in peritoneal dialysis patients. DESIGN: Prospective study. SETTING: Teaching hospital renal unit. SUBJECTS: Patients on regular peritoneal dialysis. INTERVENTIONS: Whole body and segmental MFBIA measurements before and after drainage of peritoneal fluid. RESULTS: Changes estimated by MFBIA in total body water (-0.4 (0.8) litres) and extracellular water (-0.3 (0.3) litres) were much lower than the actual changes (2.0 (0.4) litres), P<0.0001. Derived resistances Recf and Ricf increased significantly for the trunk but not for total body measurements and changes did not correlate with volumes of fluid drained. CONCLUSIONS: MFBIA is limited in its ability to detect intraperitoneal fluid, using both whole body and segmental techniques. PMID- 10822297 TI - Prokaryotic gene therapy to combat multidrug resistant bacterial infection. PMID- 10822298 TI - Intramuscular administration of E7-transfected dendritic cells generates the most potent E7-specific anti-tumor immunity. AB - Dendritic cells (DCs) are highly efficient antigen-presenting cells capable of priming both cytotoxic and helper T cells in vivo. Recent studies have demonstrated the potential use of DCs that are modified to carry tumor-specific antigens in cancer vaccines. However, the optimal administration route of DC based vaccines to generate the greatest anti-tumor effect remains to be determined. This study is aimed at comparing the levels of immune responses and anti-tumor effect generated through different administration routes of DC-based vaccination. We chose the E7 gene product of human papillomavirus (HPV) as the model antigen and generated a stable DC line (designated as DC-E7) that constitutively expresses the E7 gene. Among the three different routes of DC-E7 vaccine administration in a murine model, we found that intramuscular administration generated the greatest anti-tumor immunity compared with subcutaneous and intravenous routes of administration. Furthermore, intramuscular administration of DC-E7 elicited the highest levels of E7-specific antibody and greatest numbers of E7-specific CD4+ T helper and CD8+ T cell precursors. Our results indicate that the potency of DC-based vaccines depends on the specific route of administration and that intramuscular administration of E7-transfected DCs generates the most potent E7-specific anti-tumor immunity. PMID- 10822299 TI - Genetic enhancement of fracture repair: healing of an experimental segmental defect by adenoviral transfer of the BMP-2 gene. AB - This study evaluated the ability of gene transfer to enhance bone healing. Segmental defects were created surgically in the femora of New Zealand white rabbits. First generation adenoviruses were used as vectors to introduce into the defects genes encoding either human bone morphogenetic protein-2 (BMP-2) or, as a negative control, firefly luciferase. Representative specimens were evaluated histologically after 8 weeks. Healing of the defects was monitored radiographically for 12 weeks, after which time the repair tissue was evaluated biomechanically. By radiological criteria, animals receiving the BMP-2 gene had healed their osseous lesions after 7 weeks, whereas those receiving the luciferase gene had not. Histologic examination of representative rabbits at 8 weeks confirmed ossification across the entire defect in response to the BMP-2 gene, whereas the control defect was predominantly fibrotic and sparsely ossified. At the end of the 12-week experiment, the control femora still showed no radiological signs of stable healing. The difference in radiologically defined healing between the experimental and control groups was statistically significant (P < 0. 002). Biomechanical testing of the femora at 12 weeks demonstrated statistically significant increases in the mean bending strength (P < 0.005) and bending stiffness (P < 0.05) of the animals treated with the BMP-2 gene. Direct, local adenoviral delivery of an osteogenic gene thus led to the healing of an osseous lesion that otherwise would not do so. These promising data encourage the further development of genetic approaches to enhancing bone healing. Gene Therapy (2000) 7, 734-739. PMID- 10822300 TI - Expression and activity of human Na+/I- symporter in human glioma cells by adenovirus-mediated gene delivery. AB - Radioiodide concentrating activity in the thyroid, mediated by human Na+/I- symporter (hNIS), provides a mechanism for effective radioiodide treatment for patients who have invasive, recurrent, and metastatic thyroid cancers after total thyroidectomy. In an attempt to develop hNIS gene transfer for radioiodide therapy for patients with brain tumors, we have constructed recombinant adenoviruses, rAd-CMV-hNIS9 and rAd-CMV-FLhNIS, to express exogenous hNIS in U1240 and U1240Tag human glioma cells. U1240Tag differs from U1240 glioma cells in that it expresses the SV40 large T antigen oncoprotein. In both U1240 and U1240Tag cells, radioiodide uptake (RAIU) activity in the cells infected with rAd CMV-hNIS9 or rAd-CMV-FLhNIS increases as the adenoviral MOI increases. The protein expression profile of hNIS in infected cells is generally in agreement with their RAIU activity profile. Although the expressed hNIS9 protein appeared to have a shorter half-life than FLhNIS, hNIS9 expression could be maintained by multiple infections in these cells. In addition, we show that hNIS can be expressed and function in a xenografted human glioma by intratumoral injection of rAd-CMV-hNIS9. PMID- 10822301 TI - Novel cell permeable motif derived from the PreS2-domain of hepatitis-B virus surface antigens. AB - Efficient transfer of proteins or nucleic acids across cellular membranes is a major problem in cell biology. Recently the existence of a fusogenic sequence was predicted in the junction area of the PreS2- and S-domain of the hepatitis-B virus surface antigens. We have identified cell permeability as a novel property of the PreS2-domain. Cell permeability of PreS2 is not restricted to hepatocytes. PreS2 translocates in an energy-independent manner into cells and is evenly distributed over the cytosol. Detailed analysis revealed that cell-permeability is mediated by an amphipatic alpha-helix between amino acids 41 and 52 of PreS2. Destruction of this translocation motif (PreS2-TLM) abolishes cell permeability. PreS2-TLM per se can act as a shuttle for peptides and functional proteins (such as EGFP). This permits the highly specific modulation of intracellular signal transduction by transfer of peptides competing protein-protein interactions as demonstrated by specific inhibition of TNFalpha-dependent activation of c-Raf-1 kinase. Moreover in vivo functionality was demonstrated by PreS2-TLM-dependent protein transfer into primary bone marrow cells and into the liver. The amphipatic motif is conserved between the different hepatitis-B virus subtypes, and the surface proteins of avian and rodent hepadnaviruses exhibit similar amphipatic peptide sequences. In respect to hepatitis-B virus-infection, the PreS2-TLM could represent the postulated fusion peptide and play a crucial role in the internalization of the viral particle. PMID- 10822302 TI - Widespread gene transfection into the central nervous system of primates. AB - We attempted in vivo gene transfection into the central nervous system (CNS) of non-human primates using the hemagglutinating virus of Japan (HVJ)-AVE liposome, a newly constructed anionic type liposome with a lipid composition similar to that of HIV envelopes and coated by the fusogenic envelope proteins of inactivated HVJ. HVJ-AVE liposomes containing the lacZ gene were applied intrathecally through the cisterna magna of Japanese macaques. Widespread transgene expression was observed mainly in the neurons. The lacZ gene was highly expressed in the medial temporal lobe, brainstem, Purkinje cells of cerebellar vermis and upper cervical cord (29.0 to 59.4% of neurons). Intrastriatal injection of an HVJ-AVE liposome-lacZ complex made a focal transfection around the injection sites up to 15 mm. We conclude that the infusion of HVJ-AVE liposomes into the cerebrospinal fluid (CSF) space is applicable for widespread gene delivery into the CNS of large animals. Gene Therapy (2000) 7, 759-763. PMID- 10822303 TI - Structural basis of DOTMA for its high intravenous transfection activity in mouse. AB - Eleven structural analogues of two known cationic lipids, N-[1-(2, 3 dioleyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTMA) and N-[1-(2,3 dioleoyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTAP) were synthesized and utilized to evaluate the structural characteristics of DOTMA for its high intravenous transfection activity. Using a CMV-driven expression system and luciferase gene as a reporter, the transfection activity of these analogues was evaluated in mice using tail vein injection. Results concerning the structure activity relationship with regard to the influence of the backbone, relative position between head group and the hydrophobic chains on the backbone, linkage bonds, as well as the composition of the aliphatic chains revealed that cationic lipids which give a higher in vivo transfection activity share the following structural characteristics: (1) cationic head group and its neighboring aliphatic chain being in a 1,2-relationship on the backbone; (2) ether bond for bridging the aliphatic chains to the backbone; and (3) paired oleyl chains as the hydrophobic anchor. Cationic lipids without these structural features had lower in vivo transfection activity. These structural characteristics, however, did not significantly influence their in vitro transfection activity. The contribution that cationic lipids make to the overall in vivo transfection activity is likely to be determined by the structure of DNA/lipid complexes and by the outcome of the interaction between the DNA/lipid complexes and blood components upon intravenous administration. PMID- 10822304 TI - A novel N-acyl phosphatidylethanolamine-containing delivery vehicle for spermine condensed plasmid DNA. AB - A unique method for formulation of plasmid DNA with phospholipids has been devised for the purpose of producing vehicles that can mediate gene delivery and transfection of living cells. The polycation, spermine, was used to condense plasmid DNA within a water-in-chloroform emulsion stabilized by phospholipids. After organic solvent removal, the particles formed could be extruded to a number average size of about 200 nm and retained DNA that was protected from nuclease digestion. This resulted in a relatively high protected DNA-to-lipid ratio of approximately 1 microg DNA/micromol lipid. The size distribution of the preparation was relatively homogeneous as judged by light microscopy and quasi elastic light scattering. Electron microscopic studies showed structural heterogeneity, but suggested that at least some of the plasmid DNA in this preparation was in the form of the previously observed spermine-condensed bent rods and toroids and was encapsulated within liposomal membranes. Preparations with the fusogenic phospholipid composition, 1, 2-dioleoyl-sn-glycero-3 phosphoethanolamine-N-dodecanoyl/ 1, 2-dioleoyl-sn-glycero-3-phosphocholine, showed transfection activity for several cells lines, particularly OVCAR-3 cells. The transfection activity sedimented with the lipid during centrifugation, confirming the association of active plasmid DNA with phospholipids. Transfection efficiency in culture was found to be of the same order of magnitude as cationic lipoplexes but much less toxic to the cells. Significant transfection of OVCAR-3 cells in tissue culture could also be observed, even in the presence of the intraperitoneal fluid from a mouse with an OVCAR-3 ascites tumor. These data indicate a new type of liposomal gene delivery system devoid of cationic lipids, phosphatidylethanolamine, cationic polymers and viral components. PMID- 10822305 TI - Tri-iodothyronine and a deleted form of hepatocyte growth factor act synergistically to enhance liver proliferation and enable in vivo retroviral gene transfer via the peripheral venous system. AB - Retroviral vectors integrate into the target cell genome in a stable manner and therefore offer the potential for permanent correction of the genetic diseases that affect the liver. These vectors, however, usually require cell division to occur in order to allow provirus entry into the nucleus. We have explored clinically acceptable methods to improve the efficiency of retroviral gene transfer to the liver, which avoid the need for liver damage. Tri-iodothyronine (T3) and recombinant hepatocyte growth factor have previously been used to induce hepatocyte proliferation in rat livers and allow in vivo retroviral gene transfer. We investigated the combined effects of these growth factors, with their differing mechanisms of action, on hepatocyte proliferation in vivo and assessed their effectiveness in priming cells for retroviral gene transfer. During the phase of hepatocyte proliferation retrovirus was administered via either the portal or tail vein. Acting synergistically, T3 and a truncated form of recombinant hepatocyte growth factor (dHGF) induced 30% of hepatocytes in normal rat liver to enter DNA synthesis at 24 h. This increased proliferation enabled the liver to be transduced in vivo by retroviral vectors via either the portal or peripheral venous system, achieving transduction efficiencies of 6.9 +/ 1.6% and 4.3 +/- 0.4% respectively. Thus, the liver can be simply and conveniently transduced in vivo with integrating vectors, introduced via the peripheral venous system during a wave of growth factor-induced proliferation, pointing the way to clinically applicable gene transfer techniques. PMID- 10822306 TI - Tumor-selective gene transduction and cell killing with an oncotropic autonomous parvovirus-based vector. AB - A recombinant MVMp of the fibrotropic strain of minute virus of mice (MVMp) expressing the chloramphenicol acetyltransferase reporter gene was used to infect a series of biologically relevant cultured cells, normal or tumor-derived, including normal melanocytes versus melanoma cells, normal mammary epithelial cells versus breast adenocarcinoma cells, and normal neurons or astrocytes versus glioma cells. As a reference cell system we used normal human fibroblasts versus the SV40-transformed fibroblast cell line NB324K. After infection, we observed good expression of the reporter gene in the different tumor cell types, but only poor expression if any in the corresponding normal cells. We also constructed a recombinant MVMp expressing the green fluorescent protein reporter gene and assessed by flow cytometry the efficiency of gene transduction into the different target cells. At a multiplicity of infection of 30, we observed substantial transduction of the gene into most of the tumor cell types tested, but only marginal transduction into normal cells under the same experimental conditions. Finally, we demonstrated that a recombinant MVMp expressing the herpes simplex virus thymidine kinase gene can, in vitro, cause efficient killing of most tumor cell types in the presence of ganciclovir, whilst affecting normal proliferating cells only marginally if at all. However, in the same experimental condition, breast tumor cells appeared to be resistant to GCV-mediated cytotoxicity, possibly because these cells are not susceptible to the bystander effect. Our data suggest that MVMp-based vectors could prove useful as selective vehicles for anticancer gene therapy, particularly for in vivo delivery of cytotoxic effector genes into tumor cells. PMID- 10822307 TI - High efficiency retroviral vectors that contain no viral coding sequences. AB - Almost all currently available retroviral vectors based on murine leukemia virus (MLV) contain one or more viral coding sequences. Because these sequences are also present in the packaging genome, it has been suggested that homologous recombination may occur between the same nucleotide sequence in the packaging genome and the vector, resulting in the production of replication competent retrovirus (RCR). Up until now, it has been difficult to completely remove viral coding sequences since some were thought to be involved in the optimum function of the retroviral vector. For example, the gag coding sequence present in almost all available retroviral vectors has been believed to be necessary for efficient viral packaging, while the pol coding sequence present in the highly efficient vector MFG has been thought to be involved in achieving the high levels of gene expression. However, we have now developed a series of retroviral vectors that are absent of any retroviral coding sequences but produce even higher levels of gene expression without compromising viral titer. In these vectors, the intron and exon sequences from heterologous cellular or viral genes are present. When compared with the well-known MLV-based vectors, some of these newly developed vectors have been shown to produce significantly higher levels of gene expression for a longer period. In an experimental system that can maximize the production of RCR, our newly constructed vectors produced an absence of RCR. These vectors should prove to be safer than other currently available retroviral vectors containing one or more viral coding sequences. PMID- 10822308 TI - Retrovirally expressed human arylsulfatase A corrects the metabolic defect of arylsulfatase A-deficient mouse cells. AB - A deficiency of arylsulfatase A (ASA) causes the lysosomal storage disease metachromatic leukodystrophy (MLD) which is characterized primarily by demyelination of the central nervous system. ASA-deficient mice develop a disease which resembles MLD in many respects and thus serve as an appropriate animal model for this disease. To establish gene therapy protocols for ASA-deficient mice, we constructed two retroviral vectors based on the murine stem cell virus. Both vectors harbor the human ASA cDNA controlled by the retroviral promoter/enhancer element, but differ by the presence or absence of a neomycin resistance gene driven by an internal promoter. A comparative analysis of the one versus the two-gene vector and an amphotropic versus an ecotropic producer cell line revealed that the amphotropic producer cell line for the one-gene vector transfers ASA overexpression to the target cells most efficiently. The human ASA encoded by this vector is correctly expressed in heterologous mouse cells and corrects the metabolic defect of transduced ASA-deficient murine cells. The constructed one-gene vector might thus be a potentially useful tool for the development of a gene-based therapy for ASA-deficient mice. Gene Therapy (2000) 7, 805-812. PMID- 10822309 TI - Peptide nucleic acid and delivery to human mitochondria PMID- 10822311 TI - Calcium antagonists, ACE inhibitors, and the risk of cancer in hypertensive patients. PMID- 10822312 TI - Do alpha blockers cause heart failure and stroke? Observations from ALLHAT. PMID- 10822313 TI - Haemorheological factors in hypertension. PMID- 10822314 TI - The accuracy of non-invasive methods for the detection of obstructive coronary artery disease in the presence of left ventricular hypertrophy. AB - In the hypertensive patient with electrocardiographic and/or echocardiographic evidence of left ventricular hypertrophy, exercise electrocardiography lacks sensitivity and specificity for the detection of obstructive coronary artery disease and should be avoided. In comparative studies both myocardial perfusion imaging and stress echocardiography are significantly more sensitive and specific for the detection of significant obstructive coronary artery disease, in the presence of left ventricular hypertrophy, and are the investigations of choice. PMID- 10822315 TI - Calcium channel blockers, ACE inhibitors, and the risk of cancer in hypertensive patients: a report from the Department of Health Hypertension Care Computing Project (DHCCP) AB - OBJECTIVE: Recent studies have shown inconsistent results on the risk of cancer in hypertensive patients using calcium channel blockers (CCBs) and angiotensin converting enzyme (ACE) inhibitors. We investigated a large number of patients from the Department of Health Hypertension Care Computing Project (DHCCP) observational database treated with these drugs for hypertension to see whether the use of CCBs for hypertension is associated with an increased risk of cancer mortality and the use of ACE inhibitors with a reduction. DESIGN: Matched case control study and a longitudinal study of survival from 1 year after presentation. PATIENTS: A total of 11663 patients treated for hypertension from 1971 through 1987. They were recruited on presentation to one of the hospital hypertension clinics or general practices involved. MAIN OUTCOME MEASURES: Death with any mention of cancer on the death certificate in patients treated with an Index drug group; CCBs, ACE inhibitors, beta adrenergic blocking drugs (BBs), or receiving a diuretic. The treatment groups were mutually exclusive. RESULTS: A total of 391 cases of cancer were matched with 1050 controls. In this case control study the adjusted relative risk estimate in comparison to diuretic treatment for CCBs was 0.79 (95% CI 0.37 to 1.69), and for CCBs plus a diuretic, 1.05 (0.65 to 1.69). Non-significant results were also observed for ACE inhibitors (1.48 (0.43 to 5.1), and 1.40 (0.56 to 3.50) with a diuretic), and also for the BB and methyldopa groups. In the longitudinal survival study, the adjusted relative risk estimate for CCBs was 1.1 (0.60 to 1.94) and 1.0 (0.53 to 1.86) for CCBs plus a diuretic, and for ACE inhibitors 1.33 (0.37 to 4.76) and 1.47 (0.67 to 3.23), respectively. CONCLUSIONS: In this population there was no increased cancer mortality with the use of CCBs and a relative risk greater than 1.7 to 2.0 was excluded with 95% confidence. The suggestion that ACE inhibitors reduce cancer mortality was not supported with best estimates of relative risk of 1.3 to 1.5 and exclusion of values less than 0.4 to 0.7. PMID- 10822316 TI - Haemorheological effects of losartan and enalapril in patients with renal parenchymal disease and hypertension. AB - The objective of this study was to compare the effects of the angiotensin II (ang II) antagonist, losartan and the angiotensin-converting enzyme inhibitor (ACEI), enalapril on haemorheology. Twenty-nine patients with renal parenchymal disease and hypertension were enrolled in the prospective, open, parallel study that involved a 14-day washout period followed by a 120-day treatment period. Patients were allocated randomly to receive either losartan 50-100 mg/day (n = 15) or enalapril 2.5-10 mg/day (n = 14) to achieve blood pressure control <140/90 mm Hg. Blood pressure, haemorheology profile and plasma fibrinogen concentration were measured after the washout phase and after 2, 10, 60, and 120 days of treatment. The data were analysed using ANOVA with repeated measures. Twenty-seven patients completed the study. Treatment with both losartan and enalapril was associated with a significant decrease (P < 0.05) in relative high shear rate whole blood viscosity, indicating an increase in blood cell deformability. In patients taking losartan, the increase in blood cell deformability did not result in a decrease in mean whole blood viscosity due to a concomitant, significant increase in mean plasma viscosity (P < 0. 01). In contrast, the improved cell deformability in patients treated with enalapril resulted in a small and statistically insignificant decrease in mean whole blood viscosity (P = 0.06; mean change = 0.15 mPa sec). The mechanism of the increase in blood cell deformability and the rise in plasma viscosity associated with losartan remain unclear. It is possible but unproven that the improvement in intrinsic blood cell rheology with losartan and enalapril may be the result of changes in cation transport systems and/or the consequence of the protective antioxidant properties of drug metabolites. PMID- 10822317 TI - High prevalence of primary aldosteronism in the Tayside hypertension clinic population. AB - Primary aldosteronism (PA) was thought to be rare but recent evidence from Australia suggests that it may be more common. As this has important implications in terms of hypertension management, we undertook to screen for this treatable condition in our hypertension clinic. We obtained blood samples in sequential patients referred for assessment in our hypertension clinic in Tayside for plasma renin activity (PRA) and aldosterone. The aldosterone to PRA ratio (ARR) was used as an initial screening test to identify potential patients with PA. Those patients with an elevated ratio (> or =750) were admitted for the salt loading and fludrocortisone suppression test. These patients also underwent adrenal CT scanning, and in selected patients, adrenal scintigraphy. Between May 1995 and January 1997 (21 months), we screened a total of 495 patients. ARR was available in 465 (93.9%) patients. Out of that number, 77 (16. 6%) had an elevated ratio of > or =750, five of whom had an adrenal adenoma (one had previous adrenalectomy). Forty-five of these patients were admitted for the salt loading and fludrocortisone suppression test with 41 positive test results suggesting PA. One patient with a negative salt loading test result however had an adenoma proven on histology. A total of 43 cases of PA were identified, giving a minimum prevalence of 9.2% (43/465). Potentially the prevalence may be up to 15% assuming that the ARR has a sensitivity of 93% (42/45) in predicting PA. In conclusion, about one in 10 patients attending a hypertension clinic may have PA. This suggests that the prevalence of PA in Tayside is as high as that in the Australian hypertensive population, and this is likely to be true elsewhere, with obvious important implications for hypertension management. PMID- 10822318 TI - The effects of exercising muscle mass on post exercise hypotension. AB - Nine recreationally active, borderline hypertensive subjects completed 30 min of arm ergometry (ARM) at 65% VO2 peak and 30 min of leg ergometry (LEG) at 70% VO2 Peak (randomised order). Blood pressure was monitored before and for 1 h after exercise using the Finapres method. Systolic, diastolic and mean blood pressures were significantly reduced for the entire 1 h post exercise. This reduction was independent of exercise modality, but there was an indication for the duration of the effect to be prolonged following the leg exercise. We conclude that the mass of the working muscle does not directly effect the magnitude of post-exercise hypotension (PEH) but may influence the duration of the response. These results suggest that a central mechanism or decreased vascular responsiveness is responsible for PEH. PMID- 10822319 TI - Antihypertensive efficacy of the ACE-inhibitor perindopril in the elderly. AB - To assess the antihypertensive efficacy of the angiotensin-converting enzyme (ACE)-inhibitor, perindopril, in the elderly, patients >65 years of age with supine diastolic blood pressure (BP) > or =90 and < or =110 mm Hg at the end of a 4-week placebo washout period were treated with perindopril 4-8 mg/daily vs placebo using a multicentre, randomised, double-blind, parallel group design. Of the 191 patients entered, 183 completed 8 weeks of double-blind therapy. Average age was 72-73 years. Supine and standing BP at the end of the placebo run-in period were 173/96 vs 168/96 mm Hg. BPs were measured in the morning, 20-25 h after the previous day's dose (ie, at the end of the dosing interval). In the placebo group, supine and standing diastolic BP decreased by 3-4 mm Hg, and systolic BP by 6-7 mm Hg. In the perindopril-group, diastolic BP decreased by 6-7 mm Hg and systolic BP by 10-13 mm Hg (both P < 0.01 vs placebo). These data indicate a substantial placebo response of particularly systolic BP in older hypertensives and indicate the importance of a parallel placebo-group to assess the extent of the actual drug's effect. Perindopril caused additional decreases in diastolic BP by about 2 mm Hg, and in systolic BP by 4-5 mm Hg. The extent of this drug-effect may be less in older vs middle-aged hypertensives. PMID- 10822320 TI - Usefulness of the I/D angiotensin-converting enzyme genotype for detecting the risk of left ventricular hypertrophy in pharmacologically treated hypertensive men. AB - The insertion/deletion polymorphism (I/D) of the angiotensin-converting enzyme (ACE) gene has been associated in some studies with a higher prevalence of left ventricular hypertrophy (LVH), but few of them were performed on pharmacologically treated hypertensive patients. The present study was undertaken to determine whether ACE genotype determination could help in the identification of pharmacologically treated hypertensive patients at a higher risk of LVH. Ninety-six consecutive men with essential hypertension were selected for the study. Left ventricular mass (LVM) was assessed by echocardiography and indexed by body surface area and 82 patients were considered suitable for the study. Three groups of patients were defined on the basis of their I/D ACE genotype: DD (n = 39), ID (n = 33) and II (n = 10). There were no statistically significant differences between the three groups regarding to the severity of hypertension at diagnosis, degree of control of blood pressure or type of antihypertensive drug therapy used. No statistically significant differences were found between the three groups regarding to LVM index (total 124 +/- 31, DD 121 +/- 29, ID 127 +/- 35 and II 122 +/- 18 g/m2), relative wall thickness (total 0.5 +/- 0. 2, DD 0.5 +/- 0.3, ID 0.48 +/- 0.07 and II 0.47 +/- 0.04) or prevalence of LVH (total 34%, DD 31%, ID 39% and II 30% by Cornell criteria and total 39%, DD 33%, ID 45% and II 40% by Framingham criteria). Furthermore, the I and D allele frequency distribution was similar in the whole group of patients, in patients with LVH, and in a control group of healthy volunteers. Our data do not support that the I/D ACE genotype determination helps in identifying treated hypertensive patients at higher risk of LVH. Journal of Human Hypertension (2000) 14, 327-331 PMID- 10822321 TI - Alterations of plasma calcium and intracellular and membrane calcium in erythrocytes of patients with pre-eclampsia. AB - BACKGROUND: Changes in plasma and intracellular calcium levels have been suggested in the pathogenesis of pre-eclampsia, however, membrane calcium content has not been studied so far. We compared intracellular and membrane calcium concentrations in erythrocytes of women with pre-eclampsia, healthy pregnant woman and controls to determine a possible alteration of membrane calcium in pre eclampsia. SUBJECTS AND METHODS: Eighteen untreated, healthy pregnant woman and 16 pregnant nulliparous women with manifest pre-eclampsia were included, 25 healthy, age-matched woman served as controls. Atomic absorption spectroscopy was used for measurement of intracellular and membrane calcium content in erythrocytes and plasmalemmal preparations. RESULTS: Plasma Ca++ concentrations were significantly lower in pre-eclamptic women (1.96 +/- 0.15 mmol/l, P < 0.01, mean +/- s.e.m.) compared to healthy controls (2.43 +/- 0.14 mmol/l) or women with uncomplicated pregnancies (2.20 +/- 0.10 mmol/l). Intracellular Ca++ concentrations were not different between groups, however, membrane Ca++ content was significantly increased in the pre-eclamptic patients (1.23 +/- 0.36 micromol/g membrane protein, P < 0.01) compared to control subjects (0.83 +/- 0.16 micromol/g) and healthy pregnant women (0.77 +/- 0.13 micromol/g). CONCLUSION: Membrane calcium content is significantly increased in pre-eclamptic women despite low plasma Ca++ concentrations. This finding suggests an altered membrane ion transport and may be of importance for the pathogenesis of pre eclampsia. PMID- 10822322 TI - Absence of sympathetic overactivity in Afro-Caribbean hypertensive subjects studied by heart rate variability. AB - Black hypertensives present a greater prevalence of left ventricular hypertrophy and an increased mortality compared to white hypertensives. Differences in sympathetic activity might contribute to explain these racial differences in hypertension. Nevertheless, previous laboratory studies did not show any increase of sympathetic activity direct to the heart in black subjects. The aim of the present study was to investigate the cardiac sympatho-vagal balance in black and white hypertensives analysing heart rate variability, during the entire 24 h. We analysed Holter recordings of 52 essential hypertensive patients, who had never received antihypertensive treatment, 26 of whom were black and 26 were white. Consecutive series of 300 beats, with 150 beats overlapped (approximately 600 series/day), were considered for the analysis in time and frequency domain. The mean 24-h value of the power of the low frequency spectral component (0.04-0.15 Hz), expressed in normalised units, ie a marker of sympathetic modulation, was significantly lower in the group of black patients compared to whites (respectively 40.0 +/- 2.1 vs 53.6 +/- 3.6 nu, P < 0.01). Similar results were observed for the LF/HF ratio, an index of the sympatho-vagal balance (respectively 4.11 +/- 0.58 vs 5.98 +/- 0.79; P < 0.05). In a multiple linear regression analysis, considering diastolic blood pressure, left ventricular mass index, race and age as independent variables, only race (P < 0.002) and age (P < 0.01) could independently predict the normalised low frequency power or the LF/HF ratio, as dependent variables. The results of this study suggest some blunting of the cardiac sympathetic neural modulation in black hypertensives compared to white hypertensives, during the entire 24 h. PMID- 10822324 TI - Studies on the pathology, especially brain hemorrhage and angioendotheliomas, induced by two new mos-containing viruses. AB - Recombinant virus 7 (R7), a spontaneous deletion mutant of SV7, which is itself a molecular clone of Moloney murine sarcoma virus 124 (MoMuSV 124), induces brain lesions and tumors of the subcutaneous tissue and spleen in all infected mice. In contrast, SV7 only induces tumors of the spleen and subcutaneous tissues. One of the genetic differences between R7 and SV7 is that R7 encodes a Gag-Mos protein whereas SV7 encodes an Env-Mos protein. To investigate whether the novel R7 gag mos oncogene is required for brain lesion induction, two viruses (SV7d1 and SVM1) were constructed in which the R7 gag-mos sequences and the adjacent 53 bp of the 5' noncoding sequence were replaced by either the SV7 or myeloproliferative sarcoma virus (MPSV) env-mos oncogenes, respectively. Like R7, SV7d1 and SVM1 induced brain lesions and tumors in the spleen and subcutaneous tissues. A prominent component of R7-, SV7d1-, and SVM1-induced tumors of the brain, subcutaneous tissues, and spleen was the presence of abnormally enlarged cells with eccentric nuclei lining vessels, scattered singly or in small clusters. Their size, localization to the luminal surface of distended vessels, and binding to Bandeiraea simplicifolia (BS-1) lectin, an endothelial cell (EC) marker, suggest that they are most likely transformed ECs. Our findings therefore indicate that the induction of brain lesions is not limited to the expression of the R7 Gag-Mos protein. However, our findings also indicate that expression of the different forms of the Mos protein results in differences in the relative abundance of ECs in brain angioendotheliomas and subcutaneous and spleen tumors induced by these viruses. PMID- 10822323 TI - Targeted foreign gene expression in spinal cord neurons using poliovirus replicons. AB - A hallmark of poliovirus is the propensity to infect and replicate in spinal cord neurons of the central nervous system. Previously, we characterized a poliovirus self-replicating RNA genome (replicon), which encodes firefly luciferase in place of the capsid genes. This replicon is encapsidated into an authentic poliovirion by providing the poliovirus capsid protein in trans. The amount of enzymatically active luciferase in cells infected with this replicon correlated with the infectious dose. To begin to characterize the in vivo infectious potential of replicons, we have inoculated mice transgenic for the human receptor for poliovirus (PVR), either intracranially or intraspinally, with the replicon encoding luciferase. Wild-type poliovirus delivered to PVR mice via intracranial or intraspinal routes resulted in paralysis and death. Replicon preparations were shown by a sensitive biological assay to be free of infectious poliovirus. Neither intracranial nor intraspinal inoculation of the replicon encoding luciferase resulted in any obvious paralysis or disease symptoms. Following intraspinal inoculation with replicons encoding luciferase, luciferase enzyme activity was detected at 4 h post-inoculation, with peak activity at approximately 8 h post-inoculation; by 48 - 72 h, the luciferase activity had returned to background levels. Luciferase activity was detected in spinal cord predominantly near the site of inoculation, although activity was detected anterior and posterior to the site of inoculation, indicating that replicons undergo limited movement within the CNS presumably via the cerebrospinal fluid. In stark contrast to poliovirus though, inoculation of replicons into the spinal cords of PVR mice did not result in noticeable pathogenesis. Using immunofluorescence with antibodies to double-stain for replicons and neurons, we determined that replicons exclusively infect the neurons of the spinal cord, with the expression of the luciferase and replicon proteins confined to the cytoplasm of the infected cells. Replicons, then, possess the identical capacity for infection of spinal cord neurons in vivo as poliovirus. The lack of discernible neuronal destruction following replicon inoculation into the spinal cord suggests that some of the pathogenesis observed during a poliovirus infection might not be due entirely to primary infection of neurons. Finally, the results of this study point to future use of replicons as a means to target recombinant protein expression to neurons in the spinal cord. PMID- 10822325 TI - Measles virus-specific T helper 1/T helper 2-cytokine production in subacute sclerosing panencephalitis. AB - Live measles virus-specific T helper 1/T helper 2-cytokine productions by peripheral blood mononuclear cells in response to live measles, mumps or varicella virus were measured in 15 patients with subacute sclerosing panencephalitis and 15 controls by enzyme-linked immunosorbent assays. Most patients with subacute sclerosing panencephalitis had a defect in measles virus specific production of interferon-gamma, one of the T helper 1 type cytokines, despite persistent presence of measles virus, with preserved interleukin-10 (T helper 2 type cytokine) synthesis. Patients with subacute sclerosing panencephalitis were divided into two groups: responders (group A) with significant interferon-gamma production (>20 pg/mL) in response to live measles virus and non-responders (group B) with a little or no interferon-gamma production. Comparison of the clinical courses between groups A and B revealed that all the patients of group A retained receptive function for a long time, while most patients of group B lost the function rapidly (P<0.01). An inverse correlation between interferon-gamma production by peripheral blood mononuclear cells and disease progression suggested that interferon-gamma plays an antiviral role in subacute sclerosing panencephalitis. PMID- 10822326 TI - JC virus binds to primary human glial cells, tonsillar stromal cells, and B lymphocytes, but not to T lymphocytes. AB - The human polyomavirus, JCV, is the etiological agent of the fatal central nervous system demyelinating disease, progressive multifocal leukoencephalopathy (PML). In PML patients, JC Virus (JCV) can be detected in glial cells in the central nervous system (CNS); in B-lymphocytes in the peripheral blood, bone marrow, spleen, and tonsil; and in tonsillar stromal cells. In vitro, JCV infects glial cells, tonsillar stromal cells, and to a limited extent B-lymphocytes. The presence or absence of as yet unidentified cell type specific transcription factors contributes to the restricted tropism of JCV for these cell types. However, several studies indicate that cell surface receptors may also contribute to the limited host range of JCV. To examine this latter possibility we measured the binding of purified JCV virions to primary cultures of glial cells, tonsillar stromal cells, peripheral blood lymphocytes, and to several established cell lines. Our results demonstrate that JCV binds to primary glial cells, stromal cells, and B cells, but does not bind to primary T cells. In contrast, JCV bound to all cell lines tested, including the Namalwa B cell line and the Jurkat T cell line. These data are novel and demonstrate that JCV selectively interacts with cells in vivo that are known to be susceptible to infection. This selectivity appears to be lost when one examines virus binding to a variety of human, monkey, or mouse tumor cell lines. We next examined the susceptibility of primary peripheral blood lymphocytes and the Namalwa B cell line to infection with JCV. Our results demonstrate that the majority of infectious JCV virions remain cell surface associated and do not efficiently establish infection of B cells. This may explain the in vivo observation that JCV DNA is frequently detected in association with lymphocytes by PCR but that JCV mRNA is rarely detected in association with lymphocytes by reverse transcriptase PCR. These results also confirm previous data regarding the association of JCV with human B cells in vivo and support the hypothesis that B cells may be involved in trafficking of JCV to the CNS. PMID- 10822327 TI - Characteristics of scrapie isolates derived from hay mites. AB - Previous epidemiological evidence suggested that in some instances a vector and/or reservoir is involved in the occurrence and spread of transmissible spongiform encephalopathies (TSEs). In a preliminary study, hay mite preparations from five Icelandic farms with a history of scrapie were injected into mice, and some of these mice became sick after long incubation periods. To confirm that the disease was scrapie, subsequent passages in mice were performed. In addition, the characteristics of the disease process in these passages were assessed and the results compared to those findings with standard scrapie strains. As expected for scrapie, subsequent passages in the same host led to shortened incubation periods compared to those in primary isolate mice, and all mice had spongiform changes in brain. Results were similar for three of four isolates with regard to clinical manifestations, the incubation periods in mice of the three scrapie incubation period genotypes (s7s7, s7p7, p7p7), and the PrPSc Western blot (WB) pattern. The characteristics of the fourth isolate were markedly different from the other three isolates with regard to these parameters. Comparison of the characteristics of standard mouse-adapted scrapie strains and the four isolates revealed differences; these differences were particularly pronounced for the fourth isolate. PMID- 10822328 TI - Detection of the human immunodeficiency virus regulatory protein tat in CNS tissues. AB - Neuropathologically, human immunodeficiency virus (HIV) is associated with a range of inflammatory disorders, extensive cortical neuronal loss, and dendritic and synaptic damage. Although the mechanisms resulting in these abnormalities are still unclear, the neurotoxic effects are thought to be due in part to viral products including the tat gene product. We have previously shown that Tat when presented to neurons extracellularly interacts with neuronal cell membranes to cause neuronal excitation and toxicity in fmole amounts. To determine the role of Tat in mediating HIV encephalitis (HIVE), we detected tat mRNA and protein in tissue extracts of nine patients with HIVE and seven patients without HIVE. Despite long autopsy times and significant degradation, tat mRNA was detected in 4/9 patients with HIVE but not in any of the seven patients without dementia. Similarly, the env mRNA was also detected in 5/9 patients with HIVE but not in the patients without HIVE. However, vif mRNA was detected in both groups of patients with (5/9) or without (2/7) HIVE. Using protein extracts from the brains of the same groups of patients we were unable to detect Tat by enzyme linked immunosorbant assay (ELISA) (sensitivity of 2 ng Tat/ml of brain tissue). However, Tat could be detected immunohistochemically and in protein extracts from the brains of rhesus macaques with encephalitis due to a chimeric strain of HIV and simian immunodeficiency virus (SHIV). Our observations support the role of Tat in the neuropathogenesis of HIV and SHIV encephalitis. PMID- 10822329 TI - Increased activity of matrix metalloproteinases in the cerebrospinal fluid of patients with HIV-associated neurological diseases. AB - Matrix metalloproteinases (MMPs) have been identified as mediators of brain injury in HIV-associated neurological diseases. The activity of the 72 kDa gelatinase A (MMP-2) and 92 kDa gelatinase B (MMP-9) was detected by zymography in the cerebrospinal fluid (CSF) of 138 HIV-infected patients (40 with AIDS dementia, 83 with brain opportunistic infections and 15 neurologically asymptomatic), 26 HIV-seronegative individuals with inflammatory neurological diseases (IND) and 12 HIV-seronegative subjects with noninflammatory neurological diseases (NIND). MMP-2 was present in all CSF samples from HIV-seropositive and HIV-seronegative individuals, including those of subjects with NIND. On the contrary, MMP-9 was absent in the CSF of NIND controls, whereas the activity of this MMP was found in the 77 - 100% of CSF samples from HIV-infected patients, including those with HIV dementia, central nervous system (CNS) opportunistic infections or neurologically asymptomatic subjects. The highest levels of MMP-9 were found in the CSF of patients with cryptococcosis, cytomegalovirus encephalitis and tuberculous meningitis and were comparable with those found in the CSF of HIV-negative patients with multiple sclerosis or meningitis. A significant correlation between CSF MMP-9 activity and CSF cell count was found only in patients with HIV dementia. The increased CSF activity of MMPs capable to degrade components of the extracellular matrix of blood-brain barrier may contribute to the transendothelial migration of virus-infected cells into the CNS and development of HIV-associated neurologic damage. PMID- 10822331 TI - General summary PMID- 10822330 TI - PCR detection of host and HIV-1 sequences from archival brain tissue. AB - Mutations in CCR5 and CCR2b have been recently shown to affect disease progression towards AIDS. A role for these host genotypes in AIDS dementia complex (ADC) has also been postulated but remains unclear. Additionally, brain derived envelope sequences from HIV-1 have been associated with ADC but their specific contribution to pathogenesis remains uncertain. This study demonstrates the successful use of PCR techniques to isolate host CCR5 and CCR2b, and HIV-1 V3 sequences from paraffin embedded tissues from patients with and without ADC. PCR amplification from archival tissue offers a novel approach for studying the interactions between potential neuroprotective elements in the host and virulence determinants in HIV that may contribute to differences in susceptibility to ADC. PMID- 10822332 TI - Agonist selective mu-opioid receptor trafficking in rat central nervous system. PMID- 10822333 TI - Opiate drugs: 'guilt by association'. PMID- 10822334 TI - The serotonin transporter and clozapine response. PMID- 10822336 TI - Strauch et al reply PMID- 10822335 TI - Analysis of parental-origin effects in linkage data. PMID- 10822337 TI - CRF-hyperresponsiveness, pediatric ophthalmic surgeries and adult presentation of generalized anxiety disorders: a clinical hypothesis. PMID- 10822338 TI - No association between (AAT)n repeats in the cannabinoid receptor gene (CNR1) and heroin abuse in a Chinese population. PMID- 10822339 TI - Do susceptibility loci contribute to the expression of more than one mental disorder? A view from the genetics of Alzheimer's disease. AB - The susceptibility of developing most major psychiatric disorders is determined in part by contributions from risk alleles at multiple genetic loci. The central aim of this article is to highlight evidence from studies of neurodegenerative disorders suggesting that some of these alleles are shared by more than one psychiatric disorder, and to explore mechanisms that may underly these pleiotropic effects. The identification of constellations of susceptibility alleles associated with particular mental disorders will provide opportunities for new insights into the molecular and cellular pathophysiology of these disorders, and will have a major impact on psychiatric research and clinical care. This approach to reducing the variance in etiopathogenesis is also likely to be important for achieving the optimal use of available treatments (maximizing effectiveness and minimizing side effects), and for the discovery of novel medications or other interventions. PMID- 10822340 TI - Effects of a novel corticotropin-releasing-hormone receptor type I antagonist on human adrenal function. AB - Corticotropin-releasing hormone (CRH) is the principal regulator of the hypothalamic-pituitary-adrenal (HPA) axis and an activator of the sympathoadrenal (SA) and systemic sympathetic (SS) systems. Mental disorders, including major depression and, more recently, Alzheimer's disease have been associated with dysregulation of the HPA axis and the SA/SS systems. Treatment of rats or monkeys with the novel CRH receptor type 1 (CRH-R1) antagonist antalarmin inhibits the HPA and/or the SA/SS axes. This is the first study to examine the potential direct effect of antalarmin on human adrenal function. Adrenocortical and adrenomedullary cells were characterized by double-immunohistochemistry with anti 17 alpha hydroxylase (cortical cells) and anti-chromogranin A (chromaffin cells). Expression of CRH, ACTH, CRH type I and type II receptor mRNA were analyzed by reverse-transcription (RT) PCR. Human adrenal cortical and/or chromaffin cells in co-culture were incubated with CRH, antalarmin, and both CRH and antalarmin in vitro. Exposure of these cells to corticotropin or vehicle medium served as positive and negative controls, respectively. Cortical and chromaffin tissues were interwoven in the human adrenals, and both in situ and in the co-culture system the endocrine cell types were in close cellular contact. ACTH, CRH, and CRH-R1 and CRH-R2 mRNAs were expressed in the human adrenal as determined by RT PCR. CRH (10-8 M) led to a moderate increase of cortisol release (145.7 +/- 20.0%) from cortical and chromaffin adrenal cells in co-culture. This effect corresponded to 41.8% of the maximal increase induced by ACTH (10-8 M). The action of CRH was completely inhibited by antalarmin. CRH, ACTH, and both CRH-R1 and CRH-R2 mRNAs are expressed in the adult human adrenal gland. CRH stimulates cortisol production in cortical and chromaffin cell co-cultures. This effect is blocked by antalarmin, a selective CRH-R1 receptor antagonist, suggesting that CRH-R1 receptors are involved in an intraadrenal CRH/ACTH control system in humans. PMID- 10822341 TI - Disease-specific alterations in frontal cortex brain proteins in schizophrenia, bipolar disorder, and major depressive disorder. The Stanley Neuropathology Consortium. AB - Severe psychiatric disorders such as schizophrenia, bipolar disorder and major depressive disorder are brain diseases of unknown origin. No biological marker has been documented at the pathological, cellular, or molecular level, suggesting that a number of complex but subtle changes underlie these illnesses. We have used proteomic technology to survey postmortem tissue to identify changes linked to the various diseases. Proteomics uses two-dimensional gel electrophoresis and mass spectrometric sequencing of proteins to allow the comparison of subsets of expressed proteins among a large number of samples. This form of analysis was combined with a multivariate statistical model to study changes in protein levels in 89 frontal cortices obtained postmortem from individuals with schizophrenia, bipolar disorder, major depressive disorder, and non-psychiatric controls. We identified eight protein species that display disease-specific alterations in level in the frontal cortex. Six show decreases compared with the non-psychiatric controls for one or more diseases. Four of these are forms of glial fibrillary acidic protein (GFAP), one is dihydropyrimidinase-related protein 2, and the sixth is ubiquinone cytochrome c reductase core protein 1. Two spots, carbonic anhydrase 1 and fructose biphosphate aldolase C, show increase in one or more diseases compared to controls. Proteomic analysis may identify novel pathogenic mechanisms of human neuropsychiatric diseases. PMID- 10822343 TI - Histamine H2 receptor gene variants: lack of association with schizophrenia. AB - A role of histaminergic neuronal systems in schizophrenia was suggested by an association with several polymorphisms located in the coding region of the histamine H2-receptor (H2R) gene (Orange et al, Mol Psychiatry 1996; 1: 466-469). Using either the reference method of direct sequencing or restriction endonuclease digestion of PCR products amplified from DNA, we could not confirm the existence of these polymorphisms in 53 Swedish controls, 52 French controls and 88 French schizophrenics. In contrast, we detected a G543A transition in the coding region of the gene that was not found in the British population. This allelic variation, which was observed in 15% of the controls with no homozygotes, did not change the amino acid sequence of the receptor. We also analyzed a 1.8-kb nucleotide sequence of the promoter region in which we detected two additional polymorphisms that may modulate the expression of the H2R gene. The first one was a A-592G transition located in the minimal promoter of the gene and found in;10% of controls with no homozygotes. The second one was a G-1018A transition located in an enhancer element of the promoter and was found in;20-30% of controls (with;2-4% homozygotes). DNA analysis of the 88 French schizophrenic subjects revealed that the incidence of the three polymorphisms was not significantly different in this population. In conclusion, the present findings may suggest a surprisingly high variability of the H2R gene polymorphisms in different geographical areas but do not support an association of these allelic variations with schizophrenia. PMID- 10822342 TI - Decreased in vitro production of interferon-gamma and interleukin-2 in whole blood of patients with schizophrenia during treatment. AB - A pattern of aberrations in the T-cell cytokine system that is typical for autoimmune disorders has also been reported in patients with schizophrenia, namely a decreased interleukin-2 (IL-2) production and increased levels of the soluble IL-2 receptor (sIL-2R). It has also been reported that the production of interferon-gamma (IFN-gamma) may be lowered. In a longitudinal design, we studied the production of both IFN-gamma and IL-2 and their correlation in patients with schizophrenia during treatment and investigated whether associations exist between cytokine production and clinical variables. The production of IFN-gamma and IL-2 was measured in equal numbers (n = 29) of patients with schizophrenia (DSM-IV) and controls who were matched for age and gender. Patients were measured 1 day after admission (T1), after 14 (T2) and 28 (T2) days of treatment. Psychopathology was assessed after these times. The production of both IFN-gamma and IL-2 was significantly lower in patients than in controls throughout the whole investigation period (T1-T3). The productions of both cytokines were significantly correlated in controls (r = 0.60, P A, and a T --> C silent mutation in the third nucleotide of codon 53 in exon 2. By Fisher's exact test the silent mutation showed a trend for association (P = 0.051) with bipolar disorder suggesting that further scrutiny of this gene is warranted. PMID- 10822345 TI - A human myo-inositol monophosphatase gene (IMPA2) localized in a putative susceptibility region for bipolar disorder on chromosome 18p11.2: genomic structure and polymorphism screening in manic-depressive patients. AB - For several decades, lithium has been the drug of choice in the long-term treatment of manic-depressive illness, but the molecular mechanism(s) mediating its therapeutic effects remain to be determined. The enzyme myo-inositol monophosphatase (IMPase) in the phospholipase C signaling system is inhibited by lithium at therapeutically relevant concentrations, and is a candidate target of lithium's mood-stabilizing action. Two genes encoding human IMPases have so far been isolated, namely IMPA1 on chromosome 8q21. 13-21.3 and IMPA2 on chromosome 18p11.2. Interestingly, several studies have indicated the presence of a susceptibility locus for bipolar disorder on chromosome 18p11.2. IMPA2 is therefore a candidate for genetic studies on both etiology and lithium treatment of manic-depressive illness. Here we report that the genomic structure of IMPA2 is composed of eight exons, ranging in size from 46 bp to 535 bp. The promoter region contains several Sp1 elements and lacks a TATA-box, features typical for housekeeping genes. By a preliminary polymorphism screening of exons 2-8 in a sample of 23 Norwegian bipolar patients, we have identified nine single nucleotide polymorphisms (SNPs). Seven of the polymorphisms were located in the introns, one was a silent transition in exon 2 (159T>C) and one was a transition in exon 5 (443G>A) resulting in a predicted amino acid substitution (R148Q). Our data show that even in a small sample of bipolar patients, several variants of the IMPA2 gene can be identified. IMPA2 is therefore an intriguing candidate gene for future association studies of manic-depressive illness. PMID- 10822346 TI - Interaction between the serotonin transporter gene and neuroticism in cigarette smoking behavior. AB - Cigarette smoking behavior is influenced by both personality traits and inherited factors. Previous research showed that neuroticism-a broad personality domain that includes anxiety, depression, impulsiveness and vulnerability-increases the risk of being a smoker, primarily because of difficulty in quitting. Neuroticism has also been associated with the 5-HTTLPR, a functional polymorphism in the promoter for the serotonin transporter gene. We used population and family-based methods to analyze the joint effects of the 5-HTTLPR and neuroticism on smoking behavior in a population of 759 never, current, and former smokers, all members of sib-pairs. Our main finding is that smoking behavior is influenced by an interaction between neuroticism and 5-HTTLPR genotype. Specifically, neuroticism was positively correlated with current smoking and negatively associated with smoking cessation in individuals and siblings with poorly transcribed 5-HTTLPR-S genotypes, but not in those with the more highly expressed 5-HTTLPR-L genotype. Individuals with both a 5-HTTLPR-S genotype and a high level of neuroticism had the greatest difficulty in quitting smoking. These data, if replicated, suggest that smoking behavior is more strongly influenced by the combination of the serotonin transporter gene and neuroticism than by either factor alone, and that personality scores and 5-HTTLPR genotype may predict the clinical efficacy of certain smoking cessation drugs. PMID- 10822347 TI - Interacting effects of the serotonin transporter gene and neuroticism in smoking practices and nicotine dependence. AB - Individual differences in propensity to nicotine dependence appear to be mediated, in part, by genetic factors.1 The serotonin transporter gene has a functional polymorphism (5-HTTLPR) which modulates gene transcription and reuptake.2,3 A possible role in nicotine dependence is suggested by a link between 5-HTTLPR and neuroticism,4 a personality trait which has been related to smoking practices.5 In a cross-sectional study of 185 smokers, we utilized multiple linear regression modeling to examine the interacting effects of the 5 HTTLPR and neuroticism on smoking practices and nicotine dependence. Genotype was classified according to the presence or absence of the short (s) allele vs the long (l) allele of 5-HTTLPR (ie, s/s or s/l vs l/l). Models controlled for gender, age, race, and alcohol use. The 5-HTTLPR by neuroticism interaction effect was statistically significant in the models of nicotine intake (P = 0.05), nicotine dependence (P = 0.001), and smoking motivations (smoking to reduce negative mood (P = 0.01); smoking for stimulation (P = 0.01)). The results suggested that neuroticism was positively associated with these smoking practices among smokers with 5-HTTLPR S genotypes (s/s or s/l), but not among smokers with the L genotype (l/l). The 5-HTTLPR may modify the effects of neuroticism on smoking motivations and nicotine dependence. Assessment of 5-HTTLPR genotype and neuroticism may help to identify smokers who are more responsive to psychotropic medications, such as selective serotonin reuptake inhibitors (SSRIs), which are being used in smoking cessation treatment. PMID- 10822348 TI - Possible association of the short allele of the serotonin transporter promoter gene polymorphism (5-HTTLPR) with violent suicide. AB - There is abundant evidence that the serotonin (5-HT) system is modulating mood and several behavioural traits and that disturbances in the regulation of this system can be associated with severe behavioural malfunctions, as aggressive implusive and suicidal behaviour.1 Recently a functional polymorphism in the promoter region of the serotonin transporter gene (5-HTTLPR) was identified2 and the presence of one or two short alleles was associated with anxiety-related personality traits3 and several psychiatric disturbances, such as affective disorder4 or severe alcohol dependence.5 With respect to the importance of the 5 HT transporter in serotonergic transmission, we have genotyped the DNA of 58 Caucasian suicide victims (with unknown psychiatric diagnoses) and 110 healthy controls for the biallelic functional polymorphism in the 5-HTTLPR. We found a highly significant increased frequency of suicide victims being carriers of one or two short alleles (Fisher's Exact Test, two sided, P = 0.0003), which suggests that a genetically altered protein function within the serotonergic pathway might be involved in suicidality, independently from the clinical diagnosis. PMID- 10822349 TI - Glucocorticoid receptor alpha and beta isoforms are not mutated in bipolar affective disorder. AB - The periodically hyperactive hypothalamic-pituitary-adrenal (HPA) axis in bipolar affective disorders, as well as the reported changes in the binding characteristics of the glucocorticoid receptor (GR), suggest the possible involvement of the GR in the aetiopathology of this disease. This was investigated by screening the coding sequences of both GR isoforms, GRalpha and GRbeta, for the presence of mutations. As a genetic predisposition has been implicated, we included in this study bipolar patients who were siblings. By RT PCR of peripheral blood mononuclear cells from patients suffering from bipolar illness, using primers spanning the whole length of the GRalpha and GRbeta coding region and subsequent agarose gel electrophoresis, heteroduplex and sequence analyses, no GR mutations could be detected. Since glucocorticoid receptor activity can be modulated by agents other than the respective ligand (eg by growth factors, cytokines and stress signals), our results favor derangements in the modulation of GR activity by such agents and not in the primary structure of the receptor as aetiopathologic factors of bipolar disease. PMID- 10822350 TI - Muscarinic1 and 2 receptor mRNA in the human caudate-putamen: no change in m1 mRNA in schizophrenia. AB - Studies using tissue obtained at autopsy suggest that changes in cholinergic neurons could be important in the pathology of schizophrenia.1-4 We have previously reported a decrease in [3H]pirenzepine binding5 and [3H]AF-DX 384 binding6 to caudate-putamen (CP) from subjects who had schizophrenia. Under the conditions chosen, [3H]pirenzepine would predominately bind to muscarinic1 (M1) and muscarinic4 (M4) receptors,7whereas [3H]AF-DX 384 would mainly bind to muscarinic2 (M2) and M4 receptors.8 Given the relative concentrations of M1, M2 and M4 receptors in the human CP and the magnitude of the decreases in radioligand binding in schizophrenia, our results most likely reflected a change in the density of M1 and M2 receptors in the CP from the schizophrenic subjects. In situ hybridisation has now been used to determine levels of m1 and m2 mRNA in CP from 14 schizophrenic and 16 control subjects previously used for radioligand binding. m2 mRNA in the CP from the schizophrenic and control subjects was below the sensitivity of in situhybridisation. There was no difference in the levels of m1 mRNA in CP from schizophrenic and control subjects (mean +/- SEM: 103 +/- 16 vs106 +/- 17 fmol [35S]oligonucleotide probe g-1estimated tissue equivalents, P = 0.91). In conclusion, data from our radioligand binding studies show decreases in [3H]pirenzepine binding that are likely to reflect a decrease in the density of M1 receptors in CP from schizophrenic subjects. Our data in this study show the absence of a concomitant change in mRNA coding for that receptor. PMID- 10822351 TI - Comparative sequencing of the proneurotensin gene and association studies in schizophrenia. AB - Neurotensin (NT) is an endogenous tridecapetide1 cleaved from a precursor proneurotensin/ proneuromedin protein. NT localises within dopaminergic neurones in the mesocortical, mesolimbic and nigrostriatal systems1-3 and it is now clear that NT can selectively modulate dopaminergic neurotransmission.2-9 These anatomical and functional connections have led to the hypothesis that NT dysfunction might contribute to the pathogenesis of neuropsychiatric disorders in which disordered dopaminergic neurotransmission is suspected, particularly schizophrenia.3 The latter hypothesis has been supported circumstantially by the observation that central administration of NT produces effects similar to those produced by the peripheral administration of atypical antipsychotics,10,11 and more directly by studies showing levels of NT in cerebral spinal fluid (CSF) is lower in schizophrenics than in controls.12,13 To allow such hypotheses to be tested, we used denaturing high performance liquid chromatography (DHPLC)14 to identify three sequence variants in the neurotensin gene (NTS) that might alter NT structure or expression. However, using a case-control study design and a novel genotyping system based upon a primer extension protocol and HPLC detection,15 we found no evidence to support the hypothesis that variation in the proneurotensin gene contributes to susceptibility to schizophrenia. PMID- 10822352 TI - Confirmation of the association between bleomycin hydrolase genotype and Alzheimer's disease. AB - Bleomycin hydrolase (BH), a cysteine protease from the papain superfamily, is considered to be a candidate for the beta-secretase, which is presumably involved in the production of beta-amyloid peptide. The G/G genotype of BH was identified as a significant risk factor for the development of Alzheimer's disease (AD) in subjects not carrying the apolipoprotein E epsilon4 allele (apoE-epsilon4). However, this finding was recently challenged. We studied this polymorphism in a homogenous sample of German AD patients and controls. The over-representation of the G/G genotype in AD patients could be confirmed, however it was more pronounced in apoE-epsilon4 carriers. Additional studies should be undertaken to increase the confidence that the BH polymorphism is associated with AD and to explore the relationship between BH and apoE. PMID- 10822353 TI - Association and linkage of anxiety-related traits with a functional polymorphism of the serotonin transporter gene regulatory region in Israeli sibling pairs. AB - A functional polymorphism in the regulatory region of the serotonin transporter gene (5-HTTLPR) has been reported to be both associated and linked to anxiety related personality measures, although other studies have not replicated these findings. The current study examines both association and linkage of the gene to two major anxiety-related personality measures, the harm avoidance scale on the Tridimensional Personality Questionnaire and the neuroticism scale of the NEO-PI R, in a sample of 148 Israeli subjects comprising 74 same-sex sibling pairs. We replicated the reported association between the short allele and higher scores on the TPQ harm avoidance scale (P = 0.03), including the subscale of shyness (P = 0.02), and also found association in the same direction between the short allele and the NEO-PI-R neuroticism subscales of anxiety (P = 0.03) and depression (P = 0.04). Sib-pair linkage analysis, using the regression method, further supported a role of the 5-HTTLPR in anxiety-related personality traits. PMID- 10822354 TI - A polymorphic region in the human transcription factor AP-2beta gene is associated with specific personality traits. AB - Transcription factor AP-2beta is implicated in playing an important role during embryonic development of different parts of the brain, eg, midbrain, hindbrain, spinal cord, dorsal and cranial root ganglia.1,2 The gene encoding AP-2beta contains a polymorphic region which includes a tetranucleotide repeat of [CAAA] four or five times, located in intron 2 between nucleotides 12593 and 12612.3 Since the midbrain contains structures important for variables such as mood and personality, we have investigated if the AP-2beta genotype is associated with personality traits estimated by the Karolinska Scales of Personality (KSP). Identification of transcription factor genes as candidate genes in psychiatric disorders is a novel approach to further elucidate the genetic factors that, together with environmental factors, are involved in the expression of specific psychiatric phenotypes. The AP-2beta genotype and KSP scores were determined for 137 Caucasian volunteers (73 females and 64 males). The personality traits muscular tension, guilt, somatic anxiety, psychastenia and indirect aggression were significantly associated with the specific AP-2beta genotype, albeit with significant difference between genders. Based on this result the human AP-2beta gene seems to be an important candidate gene for personality disorders. Moreover, the present results suggest that the structure of the intron 2 region of the AP 2beta gene is one factor that contributes to development of the constitutional component of specific personality traits. PMID- 10822355 TI - Oral manifestations associated with human immunodeficiency virus (HIV) infection in developing countries--are there differences from developed countries? PMID- 10822356 TI - Oral inflammation and reactive species: a missed opportunity? PMID- 10822357 TI - Reactive oxygen species: a potential role in the pathogenesis of periodontal diseases. AB - The pathological events leading to the destruction of the periodontium during inflammatory periodontal diseases are likely to represent complex interactions involving an imbalance in enzymic and non-enzymic degradative mechanisms. This paper aims to review the increasing body of evidence implicating reactive oxygen species (ROS), derived from many metabolic sources, in the pathogenesis of periodontal tissue destruction. ROS are generated predominantly by polymorphonuclear leukocytes (PMN) during an inflammatory response and are regarded as being highly destructive in nature. The detection of ROS oxidation products, the elevation of iron and copper ions, which catalyse the production of the most reactive radical species, and the identification of an imbalance in the oxidant/antioxidant activity within periodontal pockets, suggests a significant role for ROS in periodontal tissue destruction. In vitro studies have shown that ROS are capable of degrading a number of extracellular matrix components including proteoglycans, resulting in the modification of amino acid functional groups, leading to fragmentation of the core protein, whilst the constituent glycosaminoglycan chains undergo limited depolymerisation. The identification and characterisation of connective tissue metabolites in gingival crevicular fluid (GCF) resulting from the degradation of periodontal tissues, notably alveolar bone, provides further evidence for a role for ROS in tissue destruction associated with inflammatory periodontal diseases. PMID- 10822358 TI - Oral lesions and conditions associated with human immunodeficiency virus infection in 300 south Indian patients. AB - BACKGROUND: Human immunodeficiency virus infection/acquired immunodeficiency syndrome (HIV/AIDS) is a major health problem in India. The National AIDS Control Organisation (NACO) of India reports a seropositivity of 25.03 per thousand for the whole country, as of October 1999. In spite of this high prevalence there are very few reports of oral lesions and conditions in Indian HIV/AIDS patients, which are important in early diagnosis and management of these patients. OBJECTIVE AND SETTING: The present report describes the oral lesions in 300 HIV positive symptomatic patients presenting to us at RAGAS-YRG CARE, a non governmental organisation in Chennai, South India, over a period of 9 months in 1998. METHOD: Lesions were diagnosed on clinical appearance using international criteria. RESULTS: Of the 300 patients 89% had acquired the infection through heterosexual contact. There were 205 males and 95 females, aged from 7 months to 72 years. Forty-seven percent of the patients were in the age group 21-30 years. CD4 counts were ascertained for 105 patients, 64 (62%) had CD4 counts < or = 200. A total of 217 (72%) of the 300 patients had some oral lesion when examined. Gingivitis (47%) and pseudomembranous candidiasis (33%) were the most common oral lesions. The other oral lesions seen were oral mucosal pigmentation (23%), erythematous candidiasis (14%), periodontitis (9%), angular cheilitis (8%), oral ulcers (3%), oral hairy leukoplakia (3%), hyperplastic candidiasis (1%), oral submucous fibrosis (2%) and one case of leukoplakia. CONCLUSIONS: Oral lesions occur commonly in HIV infection. A comprehensive oral examination may not only suggest HIV disease but may also be useful in monitoring the disease progression. This is a cost-effective procedure, which may be useful in screening large populations in developing countries like India. PMID- 10822359 TI - Factors associated with periodontitis in an HIV-infected southeast USA study. AB - OBJECTIVES: To determine the relationship of immunosuppression with measures of probing pocket depth (PPD), recession (REC), and clinical attachment level (CAL) in an HIV-infected population from North Carolina (NC), a state in the southeastern United States (USA). DESIGN: Cross-sectional study of HIV-infected adults (n = 326) treated at the University of North Carolina Hospitals. Clinical medical record review and sociodemographic interview data were collected. Median age of study participants was 37 years (range 19-67). Males comprised 78% and Blacks 60%. Analyses were limited to those who were dentate (n = 316). MAIN OUTCOME MEASURES: Main outcomes were cases vs non-cases of notable PPD, REC, and CAL. Immunosuppression measured by CD4+ cell count microL was the exposure of interest. RESULTS: Defined cases of PPD (n = 148) were 2.6 (95% CI = 1.3, 5.3) times less likely to occur at CD4+ cells < 200 than non-cases, whereas, cases of REC (n = 94) were 2.8 (95% CI = 1.2, 6.6) times more likely to occur at that level of severe immunosuppression, controlling for confounders. CONCLUSION: Sub groups of persons with HIV experience a high burden of periodontitis where notable severity and extent of PPD, CAL, and REC were clearly evident at different stages of immunosuppression. PMID- 10822360 TI - The effect of brief exposure to sub-therapeutic concentrations of chlorhexidine gluconate on the germ tube formation of oral Candida albicans and its relationship to post-antifungal effect. AB - OBJECTIVES: Adherence of Candida albicans has been implicated as the first step in the pathogenesis of oral candidosis, and germ tube formation a contributory attribute. Recently, these organisms have also been implicated in persistent apical periodontitis. Chlorhexidine gluconate is by far the commonest antiseptic mouth wash prescribe in dentistry. As the intraoral concentrations of this antiseptic fluctuate considerably due to the dynamics of the oral cavity, the main objective of this study was to investigate the effect of brief exposure to three different sub-therapeutic concentrations of chlorhexidine gluconate (0.005%, 0.0025% and 0.00125%) on the germ tube formation of C. albicans. These findings were then correlated with the chlorhexidine-induced post-antifungal effect (PAFE) values we obtained in a study using the identical organisms and experimental conditions. DESIGN: Ten oral isolates of C. albicans were exposed to three different concentrations of chlorhexidine gluconate for 30 min, the antiseptic removed, and the germ tube formation of these isolates quantified following subsequent incubation in a germ tube inducing medium. The PAFE was evaluated by turbidometric measurement of growth. RESULTS: When compared with the controls, exposure to 0.005%, 0.0025% and 0.00125% chlorhexidine gluconate suppressed the ability to form germ tubes by 81.23% (P < 0.01), 42.74% (P < 0.01) and 9.13% (P > 0.05), respectively, while eliciting a mean PAFE of 9.91 h, 1.65 h and 0.67 h respectively. On regression analysis a significant positive correlation was observed between these two parameters (P < 0.0001; r = 0.7325). CONCLUSIONS: Taken together, these findings imply that short exposure to sub therapeutic levels of chlorhexidine gluconate may modulate candidal germ tube formation as well as its growth, thereby suppressing its pathogencity in vivo. PMID- 10822361 TI - Ultrastructural study of direct bone formation induced by BMPs-collagen complex implanted into an ectopic site. AB - OBJECTIVE: Some authors have reported that direct bone formation is ectopically induced by bone morphogenetic proteins (BMPs) independently of cartilage formation when type I collagen is used as a carrier. This study ultrastructurally investigated the mechanism of direct bone formation by BMPs-collagen complex. MATERIALS AND METHODS: Partially purified BMPs were combined with atelopeptide type I collagen (AC) and were implanted into the calf muscles of rats (n = 20). Tissue specimens were removed on days 7, 10, 14, and 21 after implantation. RESULTS: Ultrastructurally, several regions near the pellet rim showed evidence of early calcification on day 10. In the uncalcified regions, mitochondrial calcification was seen in mesenchymal cells near AC fibers. The initially calcified regions contained numerous calcified granules deposited in the AC fibers. Some of these granules adhered to the cell membrane of osteogenic cells. In the highly calcified regions, some osteogenic cells secreted uncalcified matrix and deposited needle-like crystals and calcified collagen microfibrils on the AC fibers. CONCLUSION: This study suggests that the mesenchymal cells invading BMPs-AC complex closely contacted the AC fibers, differentiated into osteogenic cells, and deposited calcified matrix on the AC fibers, resulting in direct osteoinduction without cartilage formation. PMID- 10822362 TI - Structure-activity relationships of sialogogic heptapeptides analogous to physalaemin. AB - OBJECTIVES: The rationale behind this study was to determine in detail which amino acids in physalaemin are crucial to its sialogogue activity, with a view of synthesizing new sialogogues which might be of use in the treatment of dry mouth. METHODS: With the progressive elimination of amino acids, one by one, from the C- and N-terminal regions, 126 heptapeptides were newly synthesized by the multipin peptide method, for comparison with II naturally occurring tachykinins. RESULTS: The C-terminal amide in position II was essential for salivation, but not the pyrolidine group or the N-terminal amino acid residues in positions I to 4. In 18 heptapeptides in which M in position II (MII) was replaced by another amino acid, one by one, none caused salivation. In 18 heptapeptides, in which L10 or G9 was replaced, three peptides caused salivation but none had significantly increased secretory activities. In 18 heptapeptides in which Y8 was replaced, four caused salivation but only one (I) had significantly increased secretory activity. In 18 heptapeptides in which F7 was replaced, only Y caused salivation but with significantly reduced secretory activity. In contrast, in 18 heptapeptides in which K6 and N5 were replaced, most caused salivation and some of them had significantly increased secretory activities. CONCLUSIONS: It is concluded that the sequence FYGLM-NH2 conserved in the C-terminal region of physalaemin is optimal, that amides in position II and F7 are very important for salivation, but that K6 and N5 can be replaced by some other amino acids, resulting in increased secretory activities. PMID- 10822363 TI - Abstracts of papers presented at the 24th Annual Meeting of the British Society for Oral and Maxillofacial Pathology, University of Leeds: Friday 10th September 1999. PMID- 10822364 TI - 'Oral cancer'--the terminology dilemma. PMID- 10822365 TI - Plasma HIV-1 RNA and route of transmission in oral candidiasis and oral hairy leukoplakia. PMID- 10822366 TI - Caspase-dependent cleavage of the retinoblastoma protein is an early step in neuronal apoptosis. AB - Rb-deficient embryos (Rb-/-) show abnormal degeneration of neurons and die at mid gestation, suggesting that RB may protect against apoptosis. Having previously shown that cyclin D1 accumulates during K+-induced apoptosis of granule neurons, we chose to investigate the role of RB under these conditions. We show that RB is cleaved in its C-terminus during the onset of neuronal apoptosis. Caspase 3-like activity increases following K+ deprivation and the time course correlates with RB cleavage and apoptosis. Although the use of a specific caspase 3-like inhibitor (z-DEBD.fmk) delays RB cleavage and reduces DNA fragmentation, data implicate other caspases in these processes. However, K+ deprivation induces a gradual production of the active p20 subunit of caspase 3 (CPP32) that coincides with RB disappearance at the cellular level. Nuclear detection of a transfected HA-tagged caspase cleavage-resistant RB mutant (DEAG/D to DEAA/D) revealed a significant decrease in apoptosis of neurons expressing the RB mutant (less than 5%) relative to the wild type form of RB (40%) during K+ deprivation. Taken together, these data show that caspase-dependent cleavage of RB is an early permissive step of the apoptosis-inducing signaling pathway in neurons. They indicate a major role of RB in neuronal protection. PMID- 10822367 TI - Regulation of CDC25B phosphatases subcellular localization. AB - The CDC25B dual specificity phosphatase is involved in the control of the G2/M transition of the cell cycle. Subcellular localization might represent an important aspect of the regulation of its activity. We have examined in transiently transfected asynchronous HeLa cells the localization of HA-tagged CDC25B proteins and found that they are nuclear or cytoplasmic suggesting the existence of an active shuttling. Accordingly, localization analysis of deletion and truncation proteins indicates that CDC25B contains a putative nuclear localization signal located between residues 335 and 354. We also demonstrated that a short 58 residues deletion of the amino-terminus end of CDC25B is sufficient to retain it to the nucleus. Mutational analysis indicates that a nuclear export sequence is located between residues 28 and 40. In addition, treatment of the cells with the exportin inhibitor, Leptomycin B, has the same effect. The mutation of Ser-323, a residue that is essential for the interaction with 14-3-3 proteins, also abolishes cytoplasmic staining. The subcellular localization of CDC25B is therefore dependent on the combined effects of a nuclear localization signal, a nuclear export signal and on the interaction with 14-3-3 proteins. PMID- 10822368 TI - Induction of human Cdc37 in prostate cancer correlates with the ability of targeted Cdc37 expression to promote prostatic hyperplasia. AB - The Cdc37 gene encodes a 50 kDa protein which targets intrinsically unstable oncoprotein kinases such as Cdk4, Raf-1, and src to the molecular chaperone Hsp90. This activity is thought to play an important role in the establishment of signaling pathways controlling cell proliferation. The budding yeast Cdc37 homolog is required for cell division and mammalian Cdc37 is expressed in proliferative zones during embryonic development and in adult tissues, consistent with a positive role in proliferation. Here we report that human prostatic tumors, neoplasias and certain pre-malignant lesions display increased Cdc37 expression, suggesting an important and early role for Cdc37 in prostatic transformation. To test the consequences of increased Cdc37 levels, transgenic mice expressing Cdc37 in the prostate were generated. These mice displayed a wide range of growth-related abnormalities including prostatic epithelial cell hyperplasia and dysplasia. These data suggest that the expression of Cdc37 may promote inappropriate proliferation and may be an important early step in the development of human prostate cancer. PMID- 10822369 TI - c-Myc antagonizes the effect of p53 on apoptosis and p21WAF1 transactivation in K562 leukemia cells. AB - c-myc protooncogene positively regulates cell proliferation and overexpression of c-myc is found in many solid tumors and leukemias. In the present study we used the K562 human myeloid leukemia cell line as a model to study the functional interaction between c-Myc and p53. Using two different methods, we generated K562 transfectant cell lines with conditional expression of either c-Myc or p53. The cells expressed the p53Vall35 mutant, which adopts a wild-type conformation at 32 degrees C, while c-Myc induction was achieved with a zinc-inducible expression vector. We found that p53 in wild-type conformation induces growth arrest and apoptosis of K562. Expression of c-Myc significantly attenuated apoptosis and impaired the transcriptional activity of p53 on p21WAF1, Bax and cytomegalovirus promoters. The impairment of p21WAF1 transactivation by c-Myc was confirmed by transfection of a c-Myc-estrogen receptor fusion protein and by induction of c myc by zinc in transfected cells. Also, p53-mediated up-regulation of p21WAF1 mRNA protein were significantly reduced by c-Myc, while Bax levels were unaffected. Consistently, c-Myc increased cyclin-dependent kinase 2 activity in K562 cells expressing p53 in wild-type conformation. These results suggest that c Myc overexpression may antagonize the pro-apoptotic function of p53, thus providing a molecular mechanism for the frequently observed deregulation of c-myc in human cancer. PMID- 10822370 TI - Treatment of prostate cancer in vitro and in vivo with 2-5A-anti-telomerase RNA component. AB - Prostate cancer is the most common malignancy of elderly men in the United States. Since there is no curative treatment for advanced prostate cancer, exploration of novel modalities of treatment is essential. Telomerase, a ribonucleoprotein, is detected in the vast majority of prostate cancer, but not in normal or benign prostatic hyperplasia tissues. Thus, telomerase is expected to be a very strong candidate for targeted therapy of prostate cancer. In this study, we synthesized a 19-mer antisense oligonucleotide against the RNA component of human telomerase (hTR) linked to a 2-5A molecule (2-5A-anti-hTR) and examined its cytotoxic effect on prostate cancer cells. The 2-5A antisense strategy relies on the recruitment and activation of RNase L at the site of targeted RNA sequence. We here show that treatment with 2-5A-anti-hTR in the presence of a cationic liposome reduced cell viability of tumor cell lines tested to 9-18% within 6 days. In contrast, normal fibroblast cells were resistant to the treatment. Its effect was mainly due to induction of apoptosis by activated caspase family members. Furthermore, treatment of subcutaneous tumors in nude mice with 2-5A-anti-hTR significantly suppressed the tumor growth through induction of apoptosis (P<0.001). The treatment with 2-5A-anti-hTR may be a promising strategy for the treatment modality of prostate cancer with telomerase activity. PMID- 10822371 TI - The cytokine hepatocyte growth factor/scatter factor inhibits apoptosis and enhances DNA repair by a common mechanism involving signaling through phosphatidyl inositol 3' kinase. AB - Scatter factor (SF) [aka. hepatocyte growth factor (HGF)] (designated HGF/SF) is a multifunctional cytokine that stimulates tumor cell invasion and angiogenesis. We recently reported that HGF/SF protects epithelial and carcinoma cells against cytotoxicity from DNA-damaging agents and that HGF/SF-mediated cytoprotection was associated with up-regulation of the anti-apoptotic protein Bcl-XL in cells exposed to adriamycin. We now report that in addition to blocking apoptosis, HGF/SF markedly enhances the repair of DNA strand breaks caused by adriamycin or gamma radiation. Constitutive expression of Bcl-XL in MDA-MB-453 breast cancer cells not only simulated the HGF/SF-mediated chemoradioresistance, but also enhanced the repair of DNA strand breaks. The ability of HGF/SF to induce both chemoresistance and DNA repair was inhibited by wortmannin, suggesting that these activities of HGF/SF are due, in part, to a phosphatidylinositol-3'-kinase (PI3K) dependent signaling pathway. Consistent with this finding, HGF/SF induced the phosphorylation of c-Akt (protein kinase-B), a PI3K substrate implicated in apoptosis inhibition; and an expression vector encoding a dominant negative kinase inactive Akt partially but significantly inhibited HGF/SF-mediated cell protection and DNA repair. These findings suggest that HGF/SF activates a cell survival and DNA repair pathway that involves signaling through PI3K and c-Akt and stabilization of the expression of Bcl-XL; and they implicate Bcl-XL in the DNA repair process. PMID- 10822372 TI - cRel-TD kinase: a serine/threonine kinase binding in vivo and in vitro c-Rel and phosphorylating its transactivation domain. AB - The activity of transcription factors is often modulated by signal responsive protein kinases. Rel/NF-kappaB transcription factors are regulated by IkappaB inhibitors, the phosphorylation of which causes ubiquitination and degradation, resulting in nuclear translocation of NF-kappaB and activation of target genes. Here we report pulldown and immunoprecipitation experiments showing that a mammalian 66 kDa protein kinase binds murine c-Rel, both in vitro and in vivo. This kinase appears to have at least two binding sites on c-Rel, a proline directed serine/ threonine substrate specificity similar to MAP kinases and to specifically phosphorylate the C-terminal domain of murine c-Rel at an ERK consensus site. PMID- 10822373 TI - Evidence that de novo protein synthesis is dispensable for anti-apoptotic effects of NF-kappaB. AB - The transcription factor NF-kappaB is a positive transcription factor for a number of genes and has been recognized as an anti-apoptotic regulator. However, the mechanism by which NF-kappaB blocks apoptosis is still controversial. Here, we demonstrate the evidence that NF-kappaB could attenuate the TNF-alpha-induced apoptosis without de novo protein synthesis using human pancreatic cancer cell lines, MIA PaCa-2 and Capan-2. The TNF-alpha-induced apoptosis was blocked by IL 1beta, a potent inducer of NF-kappaB activation. This inhibitory effect of IL 1beta was evident when cells were treated with protein synthesis inhibitors such as cycloheximide (CHX). Moreover, NF-kappaB decoy oligonucleotides could not block the anti-apoptotic effect of IL-1beta at doses sufficient to block the NF kappaB-dependent transcription induced by IL-1beta. To confirm the role of NF kappaB in blocking apoptosis, we generated stable cell lines expressing IkappaBdeltaN, a highly stable form of IkappaBalpha, a cytoplasmic inhibitor of NF-kappaB. In these stable transfectants, the antiapoptotic effect of IL-1beta was totally abolished, indicating that the anti-apoptotic action of IL-1beta could be ascribed to the NF-kappaB action. These findings show that de novo protein synthesis is dispensable for anti-apoptotic effects of NF-kappaB and support the possibility that NF-kappaB could exert its anti-apoptotic action through protein-protein interaction. PMID- 10822374 TI - HTLV-1 Tax protein sensitizes cells to apoptotic cell death induced by DNA damaging agents. AB - Transient HTLV-1 Tax expression suppresses cellular nucleotide excision repair, and this effect correlates with Tax transactivation of the proliferating cell nuclear antigen promoter. The inability to repair DNA damage typically induces apoptotic cell death. Therefore, we investigated the effect of Tax-mediated suppression of DNA repair on apoptosis in stable Tax-expressing cells. Constitutive Tax expression reduced cellular nucleotide excision repair activity compared with parental and control cells. Tax-expressing cells were also more sensitive to apoptosis induced by DNA damaging agents than control cells. Even though Tax-expressing cells displayed reduced DNA repair, they showed increased DNA replication following UV damage. These results suggest that Tax suppresses the cell's ability to repair DNA damage and stimulates DNA replication even in the presence of damage. The inability to repair DNA damage is likely to stimulate apoptotic cell death in the majority of Tax-expressing cells while the ability to promote DNA replication may also allow the survival of a small population of cells. We propose that together these effects contribute to the monoclonal nature and low efficiency of HTLV-1 transformation. PMID- 10822375 TI - Somatic mutation of hPMS2 as a possible cause of sporadic human colon cancer with microsatellite instability. AB - Inactivation of DNA-mismatch repair underlies the genesis of microsatellite unstable (MSI) colon cancers. hPMS2 is one of several genes encoding components of the DNA-mismatch repair complex, and germline hPMS2 mutations have been found in a few kindreds with hereditary nonpolyposis colorectal carcinoma (HNPCC), in whom hereditary MSI colon cancers develop. However, mice bearing null hPMS2 genes do not develop colon cancers and hPMS2 mutations in sporadic human colon cancers have not been described. Here we report that in Vaco481 colon cancer the hPMS2 gene is inactivated by somatic mutations of both hPMS2 alleles. The cell line derived from this tumor is functionally deficient in DNA mismatch repair. This deficiency can be biochemically complemented by addition of a purified hMLH1 hPMS2 (hMutLalpha) complex. The hPMS2 deficient Vaco481 cancer cell line demonstrates microsatellite instability, an elevated HPRT gene mutation rate, and resistance to the cytotoxicity of the alkylator MNNG. We conclude that somatic inactivation of hPMS2 can play a role in development of sporadic MSI colon cancer expressing the full range of cancer phenotypes associated with inactivation of the mismatch repair system. PMID- 10822376 TI - Exogenous E2F expression is growth inhibitory before, during, and after cellular transformation. AB - To gain insight into the tumor suppressor properties of E2F1, we investigated growth inhibition by the E2F family of transcription factors using a tissue culture model system. We first show that exogenous E2F expression causes an 80% decrease in NIH3T3 colony formation and activated c-Ha-Ras-mediated focus formation. Inhibition of Ras-mediated transformation was dependent upon E2F DNA binding activity but did not require amino- or carboxy-terminal E2F1 protein interaction domains. Because E2F upregulation has been suggested to be associated with a neoplastic phenotype, it was possible that increased E2F activity would not be inhibitory to previously transformed cells. However, we found that exogenous E2F was also inhibitory to growth of NIH3T3 cells previously transformed by Ras or Neu. Further characterization revealed that exogenous E2F expression is inhibitory at very early times after transfection, causing dramatic losses in transfected cell populations. Interestingly, those few cells which do establish appear to be unaffected by the overexpressed E2F. Therefore, we propose that increased E2F activity may only be tolerated in a subset of cells which have acquired specific alterations that are dominant over E2F-mediated growth inhibition. PMID- 10822377 TI - PI-3-kinase is an essential anti-apoptotic effector in the proliferative response of primary human epithelial cells to mutant RAS. AB - In contrast to its growth-inhibitory effect on primary mesenchymal cells, RAS oncogene activation induces a proliferative phenotype in normal human thyroid epithelial cells in vitro, consistent with its putative role in tumour initiation. Using this model, we previously showed that activation of the MAP kinase (MAPK) pathway is necessary, but not sufficient for the proliferative response to mutant (V12) H-RAS. Here we extend this work to show that another major RAS effector-- phosphatidylinositol-3-kinase (PI-3-K)--while also insufficient alone, is able to synergize with MAPK activation to mimic the effect of mutant RAS, albeit at reduced efficiency. Furthermore we show that PI-3-K is an absolute requirement for the proliferative response to RAS in these cells, acting via suppression of RAS-induced apoptosis. These data extend our understanding of RAS signalling in a clinically-relevant cell context and point to the use of PI-3-K inhibitors as potential therapeutic agents for targetting human cancers induced by RAS mutation. PMID- 10822378 TI - Induction of S phase and apoptosis by the human papillomavirus type 16 E7 protein are separable events in immortalized rodent fibroblasts. AB - The HPV16 E7 oncoprotein neutralizes several cell cycle checkpoints, favouring the entry of quiescent cells into S phase. This activity is mediated in part by association of E7 with the pocket proteins and consequent activation of E2F transcription factors. In addition, HPV16 E7 protein is able to promote apoptosis. In this study we demonstrate that the ability to induce apoptosis is a common property of E7s belonging to both benign and malignant HPV types. The E7 induced apoptosis is mediated by inactivation of pRb, whilst neutralization of the other two pRB-related proteins, p107 and 130, is not sufficient to trigger apoptosis. Moreover, we show that certain point mutations in the conserved region 1 (CR1) of HPV16 E7 abolish the induction of apoptosis without altering the ability to stimulate S phase. Thus, these two E7-mediated cellular events, apoptosis and S phase entry, can be separated in immortalized rodent fibroblasts. Our findings demonstrate that the E7-mediated pRb destabilization is not required for its ability to drive quiescent cells into S phase and to induce apoptosis. Finally, expression of E7 proteins in NIH3T3, which lack a functional p19ARF, does not lead to p53 accumulation, indicating that the E7 impacts upon additional cellular pathways to promote apoptosis. PMID- 10822379 TI - Apoptotic crosstalk between the endoplasmic reticulum and mitochondria controlled by Bcl-2. AB - Apoptosis involves mitochondrial steps such as the release of the apoptogenic factor cytochrome c which are effectively blocked by Bcl-2. Although Bcl-2 may have a direct action on the mitochondrial membrane, it also resides and functions on the endoplasmic reticulum (ER), and there is increasing evidence for a role of the ER in apoptosis regulation as well. Here we uncover a hitherto unrecognized, apoptotic crosstalk between the ER and mitochondria that is controlled by Bcl-2. After triggering massive ER dilation due to an inhibition of secretion, the drug brefeldin A (BFA) induces the release of cytochrome c from mitochondria in a caspase-8- and Bid-independent manner. This is followed by caspase-3 activation and DNA/nuclear fragmentation. Surprisingly, cytochrome c release by BFA is not only blocked by wild-type Bcl-2 but also by a Bcl-2 variant that is exclusively targeted to the ER (Bcl-2/cb5). Similar findings were obtained with tunicamycin, an agent interfering with N-linked glycosylations in the secretory system. Thus, apoptotic agents perturbing ER functions induce a novel crosstalk between the ER and mitochondria that can be interrupted by ER-based Bcl-2. PMID- 10822380 TI - Epo regulates erythroid proliferation and differentiation through distinct signaling pathways: implication for erythropoiesis and Friend virus-induced erythroleukemia. AB - We have recently isolated the erythroleukemic cell line, HB60-5, that proliferates in the presence of erythropoietin (Epo) and stem cell factor (SCF), but undergoes terminal differentiation in the presence of Epo alone. Ectopic expression of the ets related transcription factor Fli-1 in these cells resulted in the establishment of the Epo-dependent cell line HB60-ED that proliferates in the presence of Epo. In this study, we utilized these two cell lines to examine the signal transduction pathways that are activated in response to Epo and SCF stimulation. We demonstrate that Epo, but not SCF, phosphorylates STAT-5 in both HB60-5 and HB60-ED cells. Interestingly, SCF activates the Shc/ras pathway in HB60-5 cells while Epo does not. However, both Epo and SCF are capable of activating the Shc/ras pathway in HB60ED cells. Furthermore, enforced expression of gp55 in HB60-5 cells by means of infection with the Spleen Focus Forming virus P (SFFV-P), confers Epo independent growth, which is associated with the up regulation of Fli-1. Activation of the Shc/ras pathway is readily detected in gp55 expressing cells in response to both Epo and SCF, and is associated with a block in STAT-5B tyrosine phosphorylation. These results suggest that STAT-5 activation, in the absence of Shc/ras activation, plays a role in erythroid differentiation. Moreover, Fli-1 is capable of switching Epo-induced differentiation to Epo-induced proliferation, suggesting that this ets factor regulated genes whose products modulate the Epo-Epo-R signal transduction pathway. PMID- 10822381 TI - Heterogeneities in the biological and biochemical functions of Smad2 and Smad4 mutants naturally occurring in human lung cancers. AB - Smad family members are essential intracellular signaling components of the transforming growth factor-beta (TGF-beta) superfamily involved in a range of biological activities. The loss of sensitivity to TGF-beta is frequent in human lung cancers and inactivation of Smad family members are thought to play important roles in disruption of TGF-beta signaling. In the study presented here, we characterized the biological and biochemical functions of six Smad2 and Smad4 mutants, which we previously identified in human lung cancers. All mutant Smad2 and Smad4 were in fact found to be defective in transmitting growth inhibitory signals originating from TGF-beta and incapable of activating Smad/hFAST-1 mediated transcription. Transcriptional activation of plasminogen activator inhibitor type 1 (PAI-1) was impaired in four of the six mutants due to the defects in homo- and/or hetero-oligomerization with wild-type Smads. In contrast, the remaining two Smad mutants showed a modest reduction in the PAI-1 transcriptional activation and apparently retained the ability to oligomerize with wild-type Smads. Significant loss of growth inhibition and Smad/hFAST-1 mediated transcriptional activation by all of the six mutants suggested that Smad mutants are indeed functionally impaired Smad mutations and may play a role in lung tumorigenesis. Moreover, the present findings suggest that in addition to the impairment in the homo- and/or hetero-oligomerization, there may be an alternative mechanism producing disruption of TGF-beta signaling, involving hFAST 1-or possibly other transcriptional cofactor(s)-mediated transcriptional activation. PMID- 10822382 TI - An N-terminal p14ARF peptide blocks Mdm2-dependent ubiquitination in vitro and can activate p53 in vivo. AB - The p53 tumour suppressor protein is down-regulated by the action of Mdm2, which targets p53 for rapid degradation by the ubiquitin-proteasome pathway. The p14ARF protein is also a potent tumour suppressor that acts by binding to Mdm2 and blocking Mdm2-dependent p53 degradation and transcriptional silencing. We have screened a series of overlapping synthetic peptides derived from the p14ARF protein sequence and found that a peptide corresponding to the first 20 amino acids of ARF (Peptide 3) could bind human Mdm2. The binding site for Peptide 3 on Mdm2 was determined by deletion mapping and lies adjacent to the binding site of the anti-Mdm2 antibody 2A10, which on microinjection into cells can activate p53 dependent transactivation of a reporter plasmid. To determine whether Peptide 3 could similarly activate p53, we expressed a fusion of green fluorescent protein and Peptide 3 in MCF7 and U-2 OS cells and were able to demonstrate induction of p53 protein and p53-dependent transcription. Peptide 3 was able to block in vitro ubiquitination of p53 mediated by Mdm2. Small peptides which are sufficient to block degradation of p53 could provide therapeutic agents able to restore p53 dependent cell death pathways in tumours that retain wild-type p53 expression. PMID- 10822383 TI - Frequent activation of AKT2 and induction of apoptosis by inhibition of phosphoinositide-3-OH kinase/Akt pathway in human ovarian cancer. AB - We previously demonstrated that AKT2, a member of protein kinase B family, is activated by a number of growth factors via Ras and PI 3-kinase signaling pathways. Here, we report the frequent activation of AKT2 in human primary ovarian cancer and induction of apoptosis by inhibition of phosphoinositide-3-OH kinase (PI 3-kinase)/Akt pathway. In vitro AKT2 kinase assay analyses in 91 ovarian cancer specimens revealed elevated levels of AKT2 activity (>3-fold) in 33 cases (36.3%). The majority of tumors displaying activated AKT2 were high grade and stages III and IV. Immunostaining and Western blot analyses using a phospho-ser-473 Akt antibody that detects the activated form of AKT2 (AKT2 phosphorylated at serine-474) confirmed the frequent activation of AKT2 in ovarian cancer specimens. Phosphorylated AKT2 in tumor specimens localized to the cell membrane and cytoplasm but not the nucleus. To address the mechanism of AKT2 activation, we measured in vitro PI 3-kinase activity in 43 ovarian cancer specimens, including the 33 cases displaying elevated AKT2 activation. High levels of PI 3-kinase activity were observed in 20 cases, 15 of which also exhibited AKT2 activation. The remaining five cases displayed elevated AKT1 activation. Among the cases with elevated AKT2, but not PI 3-kinase activity (18 cases), three showed down-regulation of PTEN protein expression. Inhibition of PI 3-kinase/AKT2 by wortmannin or LY294002 induces apoptosis in ovarian cancer cells exhibiting activation of the PI 3-kinase/AKT2 pathway. These findings demonstrate for the first time that activation of AKT2 is a common occurrence in human ovarian cancer and that PI 3-kinase/Akt pathway may be an important target for ovarian cancer intervention. PMID- 10822384 TI - PKC-delta is an apoptotic lamin kinase. AB - Protein kinase C-delta is activated during apoptosis, following proteolytic cleavage by caspase 3. Furthermore, overexpression of the catalytic kinase fragment of PKC-delta induces the nuclear phenotype associated with apoptosis, though the molecular basis of this effect has not been determined. In these studies we have examined the role of PKC-delta in the disassembly of the nuclear lamina at apoptosis. The nuclear lamina is disassembled during mitosis and apoptosis and mitotic disassembly involves hyperphosphorylation of lamin proteins by mitotic lamin kinases. During apoptosis, lamin proteins are degraded by caspase 6 and the contribution made by phosphorylation has not been proven. We show here that protein kinase C-delta co-localized with lamin B during apoptosis and activation of PKC-delta by caspase 3 was concomitant with lamin B phosphorylation and proteolysis. Inhibition of PKC-delta delayed lamin proteolysis, even in the presence of active caspase 6, whilst inhibitors of mitotic lamin kinases were without effect. In addition recombinant human PKC delta was able to phosphorylate lamin B in vitro suggesting that its actions are direct and not via an intermediary kinase. We propose that PKC-delta is an apoptotic lamin kinase and that efficient lamina disassembly at apoptosis requires both lamin hyperphosphorylation and caspase mediated proteolysis. PMID- 10822385 TI - Sensitization to CD95 ligand-induced apoptosis in human glioma cells by hyperthermia involves enhanced cytochrome c release. AB - CD95L-induced apoptosis involves caspase activation and is facilitated when RNA and protein synthesis are inhibited. Here, we report that hyperthermia sensitizes malignant glioma cells to CD95L- and APO2L-induced apoptosis in the absence, but not in the presence, of inhibitors of RNA and protein synthesis. Hyperthermia does not alter CD95 expression at the cell surface and does not modulate the morphology of CD95-mediated cell death on electron microscopy. Bcl-2 gene transfer inhibits apoptosis and abrogates the sensitization mediated by hyperthermia. Hyperthermia does not overcome resistance to apoptosis conferred by the viral caspase inhibitor, crm-A, indicating the absolute requirement for the activation of crm-A-sensitive caspases, probably caspase 8, for apoptosis. CD95L evoked DEVD-amc-cleaving caspase activity is enhanced by hyperthermia, suggesting that hyperthermia operates upstream of caspase processing to promote apoptosis. There is no uniformly enhanced processing of three caspase 3 substrates, poly-ADP ribose polymerase (PARP), protein kinase C (PKC) delta and DNA fragmentation factor (DFF) 45. Yet, hyperthermia promotes CD95L-evoked DNA fragmentation. Interestingly, hyperthermia enhances the CD95L-evoked release of cytochrome c in the absence, but not in the presence, of CHX. In contrast, the reduction of the mitochondrial membrane potential is enhanced by hyperthermia both in the absence and presence of CHX, and enhanced cytochrome c release is not associated with significantly enhanced caspase 9 processing. The potentiation of cytochrome c release at hyperthermic conditions in the absence of CHX is abrogated by Bcl-2. Thus, either hyperthermia or inhibition of protein synthesis by CHX potentiate cytotoxic cytokine-induced apoptosis. These pathways show no synergy, but rather redundance, indicating that CHX may function to promote apoptosis in response to cytotoxic cytokines by inhibiting the synthesis of specific proteins whose synthesis, function or degradation is temperature-sensitive. PMID- 10822386 TI - Specific inhibition of FGF-induced MAPK activation by the receptor-like protein tyrosine phosphatase LAR. AB - LAR is a widely expressed receptor-like protein tyrosine phosphatase that is implicated in regulation of intracellular signaling triggered by both cell adhesion and peptide growth factors. Genetic studies revealed that LAR regulates neuron axon path finding in Drosophila and mammary gland epithelial cell differentiation in mice. The molecular mechanism underlying the tissue specific function of LAR has not been clearly understood. We investigated the role and mechanism of LAR in peptide growth factors EGF and FGF signaling in human tissue culture cells in which the expression of LAR is under the control of an inducible promoter. We found that although both EGF and FGF induce activation of mitogen activated protein kinase (MAPK), LAR only inhibits FGF-induced MAPK activation. LAR does not interact directly with the peptide growth factor receptors, since the ligand-induced autophosphorylation of growth factor receptors was not affected by induction of LAR. The specific effect of LAR on FGF-induced MAPK activation appeared to be mediated by specific inhibition of the phosphorylation of two signal transducers that act downstream of the FGF receptor, FRS2 and a 180 kDa protein, and by prevention of their interaction with the adaptor protein GRB2. In contrast, LAR selectively inhibited the epidermal growth factor (EGF) induced phosphorylation of p130CAS and the formation of the complex between p130CAS and GRB2 but this effect did not influence the activation of MAPK by EGF. These data suggest that LAR and similar receptor-like protein tyrosine phosphatases may contribute to the regulation of transmembrane signaling by selectively inhibiting the tyrosine phosphorylation of specific signal transducers that act downstream of the plasma membrane-associated tyrosine kinases. The consequent inhibition of the formation of signaling complexes by these proteins may contribute to the specificity of the signals generated by specific peptide growth factors as well as extracellular matrix proteins. PMID- 10822387 TI - Caspases and mitochondria in c-Myc-induced apoptosis: identification of ATM as a new target of caspases. AB - The mechanism(s) of c-Myc transcription factor-induced apoptosis is still obscure. The activation of c-Myc has been found to lead into the processing/activation of caspases (caspase-3), but the significance of this for the cell demise is debatable. Here we report that several targets of caspases (PKCdelta, MDM2, PARP, replication factor C, 70 kDa U1snRNP, fodrin and lamins) are cleaved during c-Myc-induced apoptosis in Rat-1 MycER cells, indicating an important role for caspases in the apoptotic process. We further found that the ATM (ataxia telangiectasia mutated)--protein is a novel key substrate of caspases. In in vitro assays, purified recombinant ATM protein was found to be cleaved by the effector caspases 3 and 7. The functional significance of the ATM cleavage is supported by the finding that ectopic expression of ATM protected in part against apoptosis. We also show that c-Myc-induced apoptosis involves loss of mitochondrial transmembrane potential, release of cytochrome c from mitochondria into the cytosol and subsequent processing of caspase-9. The cleavage of caspase-9 is, however, minimal and a much later event than the processing/activation of caspase-3, suggesting that it is not the apical caspase. Evidence is provided that there is, nevertheless, an upstream caspase(s) regulating the functions of caspase-3 and mitochondria. Additionally, it was found that p53 becomes upregulated, together with its transcriptional targets MDM2 and p21, upon c-Myc induction, but this occurs also at a later time than the activation of caspase-3. PMID- 10822388 TI - Functional identification of the apoptosis effector BH3 domain in cellular protein BNIP1. AB - BCL-2 family proteins play a central role in apoptosis regulation in mammals and in C. elegans. Mammalian cellular and viral anti-apoptosis proteins such as BCL-2 and E1B-19K interact with several cellular proteins. Some of these interacting proteins promote apoptosis and belong to the BCL-2 family. Certain BCL-2 family proapoptotic proteins such as BAX and BAK share extensive sequence homology with BCL-2. In contrast, certain pro-apoptotic proteins such as BIK and BID share a single death effector domain, BH3, with other BCL-2 family proteins. By mutational analysis, we show that one of the cellular proteins, BNIP1 (previously Nip-1), that interacts with BCL-2 family anti-apoptosis proteins is a 'BH3 alone' pro-apoptotic protein. Transient transfection of BNIP1 induces a moderate level of apoptosis. Deletions of the N-terminal 32 amino acid region and the C-terminal trans-membrane domain did not significantly affect pro-apoptotic activity. In contrast, deletions encompassing a region containing a motif similar to the BH3 domain abrogated the apoptotic activity. Substitution of BNIP1 BH3 domain for the corresponding sequence in BAX efficiently restored the apoptotic activity of BAX, establishing the functional identity of the BH3 domain of BNIP1. The N-terminal deletions of BNIP1 (that retain the BH3 domain) enhanced the level of interaction with BCL-XL. Mutants containing the BH3 deletions were still able to heterodimerize with BCL-XL while mutants lacking both the N-terminal region and the BH3 domain were unable to heterodimerize, suggesting that BNIP1 may bind to BCL-XL via two different binding motifs. PMID- 10822389 TI - Diagnostic and prognostic value of compound motor action potential of lower limbs in acute paraplegic patients. AB - OBJECTIVES: To evaluate the diagnostic and prognostic contribution of motor nerve conduction studies (NCS) in addition to neurological examination in patients with acute paraplegia. METHODS: In 79 patients with acute onset of paraplegia due to traumatic or ischaemic damage of the conus medullaris/cauda equina (conus/cauda) or lesion of the mid-thoracic spinal cord (epiconal) neurological (initial and follow-up clinical motor and sensory scores; outcome of ambulatory capacity determined at least 6 months post-trauma) and electrophysiological examinations (motor nerve conduction velocity (MNCV) and compound motor action potential (CMAP) of tibial and peroneal nerves) were performed in parallel. RESULTS: Severe axonal motor neuropathies were significantly caused by conus/cauda lesions (loss of tibial CMAP in 71% and of peroneal CMAP in 68%) compared to patients with epiconal lesion (no loss of tibial CMAP and abolished peroneal CMAP in 14%). The CMAPs were deemed acutely pathological 4 - 14 days post-trauma and were indicative of the severity of conus/cauda lesion while the MNCV remained normal. Follow-up recordings (up to 1 year post trauma) revealed no significant change in the CMAP values. The clinical examination according to the American Spinal Injury Association (ASIA protocol) in contrast to the CMAP values was significantly related to the outcome of ambulatory capacity. CONCLUSIONS: In contrast to patients with an epiconal SCI almost all patients with damage of the conus/cauda present a severe axonal neuropathy of the tibial and peroneal nerves. Pathological CMAPs develop as early as 1 - 2 weeks after onset of acute paraplegia. They allow, at an early stage, to differentiate between conus/cauda or epiconal lesion and to assess the severity of conus/cauda lesion. Thereafter follow-up examinations remain stable and a developing worsening of peripheral nerve or spinal cord function, eg due to post-traumatic syringomyelia, may be indicated by a secondary deterioration of CMAP values. The clinical examination, according to the ASIA protocol, in acute paraplegia patients, in contrast to the motor nerve conduction studies, is of prognostic value in predicting the outcome of ambulatory capacity. PMID- 10822390 TI - The efficacy of biochemical markers in patients with ossification of posterior longitudinal ligament of the spine. AB - STUDY DESIGN: Serum levels of carboxyterminal propeptide of type I collagen (PICP), osteocalcin (OC), carboxyterminal telopeptide of type I collagen (ICTP) and urinary levels of pyridinoline (Pyr) and deoxypyridinoline (Dpyr) were measured in patients with ossification of posterior longitudinal ligament of the spine (OPLL) and age-matched control subjects. OBJECTIVES: To evaluate the efficacy of these biochemical markers of the patients with OPLL. SETTING: Department of Orthopedic Surgery, Hamamatsu University School of Medicine, Hamamatsu, Japan. METHODS: Spot urine and blood samples were obtained from 20 healthy males aged 45 - 78 (mean+/-SD; 63. 0+/-11.5) and 22 male patients with OPLL aged 46 - 77 (mean+/-SD; 59. 9+/-8.8), and serum levels of PICP, OC, ICTP and urinary levels of Pyr and Dpyr were measured. RESULTS: There were no significant difference in age, serum PICP, OC, ICTP, urinary Pyr and Dpyr levels between OPLL and control group. CONCLUSION: Neither bone formation nor bone resorption was accelerated in the patients with OPLL. PMID- 10822391 TI - Altered contractile properties of the quadriceps muscle in people with spinal cord injury following functional electrical stimulated cycle training. AB - STUDY DESIGN: A longitudinal training study. OBJECTIVES: To assess if contractile speed and fatigability of paralysed quadriceps muscles in individuals with spinal cord injury (SCI) can be altered by functional electrical stimulation leg cycle ergometry (FES-LCE) training. SETTINGS: The Sint Maartenskliniek rehabilitation centre and the University of Nijmegen, Nijmegen, the Netherlands. METHODS: Contractile properties of the quadriceps muscle were studied in seven people with motor-complete SCI who participated in a FES-LCE training program. Subjects trained for 30 min, three times per week for 6 weeks. Contractile speed and fatigue characteristics of electrically stimulated isometric contractions were compared before and after 6 weeks of FES-LCE. RESULTS: Fatigue resistance improved following FES-LCE training as indicated by the higher forces maintained in response to repetitive electrical stimulation. In contrast with an improved fatigue resistance, the maximal rate of force rise was unaffected, the speed of relaxation increased and the fusion of a 10 Hz force signal decreased. Furthermore, the force-frequency relationship shifted to the right at low stimulation frequencies, indicated by a decline in the ratio of 1 and 100 Hz force responses as well as the ratio of 10 and 100 Hz force responses. CONCLUSION: FES-LCE training can change the physiological properties of the quadriceps muscle in people with SCI. Even after a short period of training, the stimulated muscles become more resistant to fatigue. Furthermore, the increased speed of relaxation and associated decreased fusion and altered force-frequency relationship following training may be related to adaptations in the calcium handling processes, which reflect an early response of long-term disused muscles. PMID- 10822392 TI - Effect of acute heat exposure on skin blood flow of the paralyzed thigh in persons with spinal cord injury. AB - OBJECTIVES: To investigate whether increased body temperature during heat exposure promotes skin blood flow of the thigh (SBFT) in individuals with spinal cord injury at rest. STUDY DESIGN: A prospective study with high lesion and with low lesion. SETTING: Hiroshima University, Higashi-Hiroshima, Japan. METHODS: Seven male paraplegics (T5-L2) and seven able-bodied subjects volunteered to participate in the experiment. First the subject rested for 1 h at an ambient temperature (Ta) of 25 degrees C and a relative humidity (Rh) of 50%. Thereafter, the subject moved to a climatic chamber heated to Ta of 33 degrees C an Rh of 50 55% and was exposed to this environment for 1 h. SBFT, stroke volume, heart rate, the skin temperature of the medial thigh (Tthigh) and the tympanic membrane temperature (Tty) were assessed during the experiment. RESULTS: The four subjects with high lesions between Th6 and Th10 (HL) showed no increase in SBFT, while the remaining three subjects with low lesion at Th11 or Th12 (LL) showed increased SBFT in the hot environment. Although Tty increased with exposure time in heat, correlation between Tty and SBFT was not significant in either paraplegic group. However, correlation between SBFT and Tthigh was highly significant in both groups. The increase in Tthigh in LL was due to increased skin blood flow in the thigh, while in HL it was considered attributable to heat transfer from the environment to the skin. CONCLUSION: These findings indicate that SBFT during heat exposure was largely dependent on the level of spinal cord injury. SBFT with low lesions increased during heat exposure when the sympathetic cutaneous vasomotor supply of the thigh was preserved. PMID- 10822393 TI - Early autonomic dysreflexia. AB - INTRODUCTION: During the stage of spinal shock the conventional view is that autonomic activity is abolished. Here, evidence is presented that autonomic activity is still present. PATIENTS: Four patients with acute cord transactions are presented: one new case and three from the literature. DEFINITIONS: The definitions of spinal shock and autonomic dysreflexia are given. METHODS: All four cases showed acute autonomic dysreflexia between 7 and 31 days after acute cord transection at a stage when the tendon reflexes were abolished. RESULTS: Two cases showed a severe rise in blood pressure; the two earlier cases, before blood pressure was routinely recorded, profuse sweating. In two cases autonomic dysreflexia was obtained when the bladder was overdistended with 1000 ml and 1600 ml. In the other two cases it occurred in response to traumatic catheterisation. This was found when supramaximal stimuli were applied. It has not been recorded routinely as, with modern management, the bladder does not get overdistended or traumatised. DISCUSSION: Other evidence, the blood pressure, and urethral tone is presented to show that sympathetic reflex activity of the cord is not abolished during spinal shock. CLINICAL SIGNIFICANCE: The clinical importance of this is that autonomic dysreflexia can be seen at an early stage and it should be considered in the differential diagnosis of a sick patient immediately after spinal injury. PMID- 10822394 TI - Walking index for spinal cord injury (WISCI): an international multicenter validity and reliability study. AB - STUDY DESIGN: Construction of an international walking scale by a modified Delphi technique. OBJECTIVE: The purpose of the study was to develop a more precise walking scale for use in clinical trials of subjects with spinal cord injury (SCI) and to determine its validity and reliability. SETTING: Eight SCI centers in Australia, Brazil, Canada (2), Korea, Italy, the UK and the US. METHODS: Original items were constructed by experts at two SCI centers (Italy and the US) and blindly ranked in an hierarchical order (pilot data). These items were compared to the Functional Independence Measure (FIM) for concurrent validity. Subsequent independent blind rank ordering of items was completed at all eight centers (24 individuals and eight teams). Final consensus on rank ordering was reached during an international meeting (face validation). A videotape comprised of 40 clips of patients walking was forwarded to all eight centers and inter rater reliability data collected. RESULTS: Kendall coefficient of concordance for the pilot data was significant (W=0. 843, P<0.001) indicating agreement among the experts in rank ordering of original items. FIM comparison (Spearman's rank correlation coefficient=0.765, P<0.001) showed a theoretical relationship, however a practical difference in what is measured by each scale. Kendall coefficient of concordance for the international blind hierarchical ranking showed significance (W=0.860, P<0.001) indicating agreement in rank ordering across all eight centers. Group consensus meeting resulted in a 19 item hierarchical rank ordered 'Walking Index for Spinal Cord Injury (WISCI)'. Inter rater reliability scoring of the 40 video clips showed 100% agreement. CONCLUSIONS: This is the first time a walking scale for SCI of this complexity has been developed and judged by an international group of experts. The WISCI showed good validity and reliability, but needs to be assessed in clinical settings for responsiveness. PMID- 10822395 TI - Use of the NESS handmaster to restore handfunction in tetraplegia: clinical experiences in ten patients. AB - OBJECTIVE: To explore possible functional effects of the Handmaster in tetraplegia and to determine suitable patients for the system. PATIENTS: Patients with a cervical spinal cord injury between C4 and C6, motor group 0 - 3. Important selection criteria were a stable clinical situation and the absence of other medical problems and complications. DESIGN: Ten patients were consecutively selected from the in- and outpatient department of a large rehabilitation hospital in The Netherlands. Each patient was fitted with a Handmaster by a qualified therapist and underwent muscle strength and functional training for at least 2 months. METHODS: Functional evaluation comprised the performance of a defined set of tasks and at least one additional task as selected by patients themselves. Tasks were performed both with and without the Handmaster. Finally, patients were asked for their opinion on Handmaster use as well as their willingness to future use. RESULTS: In six patients a stimulated grasp and release with either one or both grasp modes (key- and palmar pinch) of the Handmaster was possible. Four patients could perform the set of tasks using the Handmaster, while they were not able to do so without the Handmaster. Eventually, one patient continued using the Handmaster during ADL at home. CONCLUSION: The Handmaster has a functional benefit in a limited group of patients with a C5 SCI motor group 0 and 1. Suitable patients should have sufficient shoulder and biceps function combined with absent or weak wrist extensors. Though functional use was the main reason for using the Handmaster, this case series showed that therapeutic use can also be considered. PMID- 10822396 TI - Dosage escalation of intravesical oxybutynin in the treatment of neurogenic bladder patients. AB - OBJECTIVE: We prospectively analyzed the dose dependent outcome and side effects of neurogenic bladder patients with intravesical application of oxybutynin at our centre. MATERIALS AND METHODS: We examined the data of 32 patients with neurogenic bladders and detrusor hyperreflexia. We registered clinical outcome, continence situation, side effects and urodynamic data of patients with (A) standard dosages of intravesical oxybutynin (0.3 mg/kg bodyweight per day) and (B) with increasing dosages in steps of 0.2 mg/kg bodyweight per day up to 0.9 mg/kg bodyweight per day. RESULTS: We examined 32 patients aged 1 to 34 years, mean age 12 years. 21/32 patients became totally continent with the dosage (A). They showed a significant (P<0.01) decrease in the median max detrusor pressure (MDP) and a significant (P<0.01) increase in the median compliance and the median age adjusted bladder capacity (AABC). Eleven out of 32 patients remained incontinent under this dosage (A). Their median MDP, their median compliance and their median AABC remained nearly unchanged. Seven out of 11 incontinent patients under dosage (A) were treated efficiently with the higher dosages (B). Their median necessary dosage escalation to achieve treatment success was 0.7 mg/kg bodyweight per day (range 0.5 to 0.9 mg/kg bodyweight per day). Their median MDP was significantly (P<0.05) decreased and their median compliance and median AABC were significantly (P<0.05) increased. Four out of 11 patients remained incontinent and showed only little improvement in urodynamic data. Two out of 11 patients with the dosage escalation (B) showed side effects at a dosage of 0. 9 mg/kg bodyweight per day. CONCLUSIONS: The intravesical application of oxybutynin was a well tolerated and efficacious therapy. The topical oxybutynin therapy dosage (A) was efficient in 66% of our selected patients, the escalating dosage titration (B) could increase the efficiency to 87%. PMID- 10822397 TI - Neurogenic colorectal dysfunction - use of new antegrade and retrograde colonic wash-out methods. AB - OBJECTIVES: To evaluate results of the Enema Continence Catheter (ECC) and the Malone Antegrade Continence Enema (MACE) applied in patients with severe neurogenic colorectal dysfunction. METHODS: The ECC was offered to 21 patients (mean age 39.9, range 7 - 72 years). The MACE was used in eight patients (mean age 32.8 years, range 15 - 66 years). All patients still using the ECC or the MACE at follow-up were interviewed. Results from patients not available for follow-up were drawn from hospital records. RESULTS: Overall success with the ECC was found in 12 of 21 patients (57%). In patients with faecal incontinence, the ECC was successful in eight out of eleven patients (73%), while four out of ten patients (40%) with constipation were successfully treated. Overall success with the MACE was found in seven out of eight patients (87%). Successful treatment with the ECC or the MACE was followed by significant improvement in quality of life. CONCLUSION: The ECC is a simple therapeutic method in severe neurogenic colorectal dysfunction. If the ECC fails the MACE, as a minor and reversible operation, is a suitable alternative to more extensive procedures. PMID- 10822398 TI - Bilateral S3 nerve stimulation, a minimally invasive alternative treatment for postoperative stress incontinence after implantation of an anterior root stimulator with posterior rhizotomy: a preliminary observation. AB - STUDY DESIGN: A preliminary report. OBJECTIVES: Urinary stress incontinence following implantation of an anterior root stimulator and a posterior rhizotomy is a rare complication which is difficult to treat. It is seen in patients with an open bladder neck (T9-L2 lesion). An artificial urinary sphincter is a possible treatment for this condition but has a higher failure rate in patients with neurogenic bladder disease and could complicate micturition. SETTING: Ghent, Belgium. METHODS: A male paraplegic patient (T9, complete lesion) aged 36 was suffering from severe urinary incontinence due to detrusor hyperreflexia. Preoperatively the bladder neck was closed on cystography. Following implantation (6/95) of an intradural anterior root stimulator with posterior rhizotomy, severe urinary stress incontinence presented. Bilateral S3 foramen leads were implanted and connected to a pulse generator. RESULTS: The patient has been continent with continuous stimulation of both S3 roots for 4 years, and no fatigue of the levator muscles has been seen. Preoperative urodynamics are compared to results 3 years postoperatively. CONCLUSION: Bilateral S3 stimulation is a feasible and minimally invasive treatment of urinary stress incontinence following implantation of an anterior root stimulator. PMID- 10822399 TI - Syringomyelia presenting as a delayed complication of treatment for nocardia brain abscess. AB - OBJECTIVE: Syringomyelia is defined as a dilatation of the central canal of the spinal cord which often leads to neurologic impairment. Syringomyelia has not previously been reported as a late complication for the treatment of brain abscess. In this report, we review a case involving this unusual association. CLINICAL PRESENTATION: A 25 year-old woman sustained a nocardia brain abscess initially presenting as a pulmonary infection. Treatment led to the development of multiloculated hydrocephalus and syringomyelia. INTERVENTION: Treatment included placement of multiple ventriculoperitoneal shunts and a syringo-pleural shunt. This resulted in stabilization of neurologic symptoms. CONCLUSION: The possibility of developing syringomyelia should be considered in any case involving post-infectious hydrocephalus. PMID- 10822400 TI - High dose methylprednisolone in the management of acute spinal cord injury - a systematic review from a clinical perspective. AB - STUDY DESIGN: Systematic literature review for primary data using predefined inclusion, exclusion and validity criteria. Primary outcome measure was standardised neurological examination or neurological function. Secondary outcomes; acute mortality, early morbidity. OBJECTIVES: To access the literature available to clinicians systematically and evaluate the evidence for an effect of high dose methylprednisolone (MPSS) on neurological improvement following acute spinal cord injury (ACSI). METHODS: Information retrieval was based on Medline search (1966 through December 1999) using the strategy 'spinal cord injury' and 'methylprednisolone' (or 'dexamethasone') with no other restrictions. Primary data publications using high dose steroids given within 12 h following spinal cord injury and reporting outcome measures separately for steroid and non-steroid treated groups were selected. Evaluation followed the guides of Guyatt et al7 (for the Evidence Based Working Group in Canada). Studies with questionable validity were excluded. Level of evidence and treatment recommendation utilised the Canadian Task Force on the Periodic Health Examination criteria.6 Experimental spinal cord injury studies on larger animals were included; small mammal experiments were considered beyond evaluation. RESULTS: Three clinical trials and six cohort study publications were found to satisfy the review criteria. The evidence they provide supports 'the recommendation that the manoeuvre (high dose methylpredisolone) be excluded from consideration as an intervention for the condition'10 (acute spinal cord injury). Twelve larger animal publications were detailed. Validity and the functional significance of results was of concern in many. The weight of evidence lay with those studies demonstrating no definite effect of MPSS on functional outcome. In cat experiments with higher level cord damage, deaths in the MPSS treated groups were notable. CONCLUSION: The evidence produced by this systematic review does not support the use of high dose methylprednisolone in acute spinal cord injury to improve neurological recovery. A deleterious effect on early mortality and morbidity cannot be excluded by this evidence. PMID- 10822401 TI - Increased V/D-ratio in lumbosacral SEP's as a new electrophysiological measure of spasticity. AB - STUDY DESIGN: Open-label, prospective study. OBJECTIVE: To establish the amplitude-ratio of V-response and D-response (V/D-ratio) as a new measure of spasticity, comparing the motor effect of the H-reflex to the sensoric input. METHODS: In 13 legs of seven patients with spasticity and in four legs of patients without central nervous system disease, maximal M-response and maximal H reflex were recorded. Lumbosacral SEP's were recorded with the same stimulus parameters as the maximal H-reflex. H/M-ratio and V/D-ratio were compared to the increased muscle tone. RESULTS: The H/M-ratio and the V/D-ratio in legs with spasticity differed significantly from the H/M-ratio and the V/D-ratio in normal legs. But only the V/D-ratio was higher in legs with moderately or highly increased muscle tone than in legs with slightly increased muscle tone. CONCLUSION: The V/D-ratio increases in spasticity and shows an even closer relationship to increased muscle tone than the H/M-ratio. PMID- 10822402 TI - Corticospinal function studied over time following incomplete spinal cord injury. AB - STUDY DESIGN: Longitudinal. OBJECTIVES: (1) To perform standard clinical neurological examinations and establish the pattern of clinical change with time following incomplete spinal cord injury (iSCI). (2) To establish the pattern of change in corticospinal electrophysiological function with time after iSCI. (3) To correlate clinical with electrophysiological findings. SETTING: The National Spinal Injuries Centre, Stoke Mandeville Hospital, Aylesbury, UK and Imperial College School of Medicine, Charing Cross Hospital, London, UK. METHODS: Neurological assessments and classification were performed according to American Spinal Injuries Association and International Medical Society of Paraplegia (ASIA/IMSOP) standards. Twenty-one patients (ages 18 - 72 years) with iSCI (level C2 - C7, ASIA impairment grades C - D) and 10 healthy control subjects (ages 27 - 57 years) were studied. Electrophysiological tests of corticospinal function were carried out using transcranial magnetic stimulation (TMS) of the motor cortex and electromyographic (EMG) recordings from thenar muscles. Both tests were performed on a number of occasions, beginning 19 - 384 days and ending 124 - 1109 days post injury, and the group data were pooled into time epochs of 50 or 100 days post injury for analysis. Seven of the patients were studied on seven or more occasions and were also assessed individually. RESULTS: Individual and pooled data indicated that neurological scores improved progressively and tended to stabilise by around 300 days post-injury. When the patients were first assessed, the mean latency for motor evoked potentials (MEPs) and inhibition of voluntary EMG were significantly different from control values. There was no significant change in latency on subsequent sessions for either the grouped or individual patient data. There was no correlation between clinical assessment and electrophysiological data. CONCLUSION: We conclude that the weakened inhibition seen following iSCI is established within a few days of the time of spinal cord trauma. We argue that reduced corticospinal inhibition may be a prerequisite for the recovery of useful motor function. SPONSORSHIP: The work was supported by a project grant from The Wellcome Trust. PMID- 10822403 TI - Clinical evaluation and management of neurogenic bowel after spinal cord injury. AB - OBJECTIVES: To assess the state of the neurological bowel in spinal cord injured (SCI) patients, design and apply a program for the comprehensive management of neurogenic bowel and evaluate outcome. SETTING: Out-patient in a Rehabilitation Service. SUBJECTS: Thirty-eight SCI patients, 12 (32%) with complete lesions of more than 5 years duration. DESIGN: Observational, longitudinal and prospective. Pre and post intervention. METHOD: Pre and post SCI intestinal function was evaluated clinically prior to beginning program. The presence of GI symptoms were studied. Laboratory work-up included colonic transit time (CTT), anorectal manometry and recto-colonoscopy. An intestinal program was designed, in order to achieve an effective and efficient evacuation in a predictable and socially acceptable time, to avoid short and long term complications and eliminate inadequate intestinal evacuation habits. OUTCOME MEASURES: Pre and post SCI difficulty in intestinal evacuation (DIE) was increased (from 2.6% to 26.3%). The most frequent GI symptom was abdominal distention (53%). Colonic inertia was present in 49% of CTT, internal anal sphincter pressure was normal or increased in 77% and rectoanal inhibitory reflex was present in 88%. With the intestinal program, the incidence of DIE was reduced to 8.8%, manual extraction (ME) was reduced from 53% to 37%. Excellent and good results were obtained in 56% of the patients. CONCLUSION: The proposed intestinal program is effective in the rehabilitation of SCI patients with neurogenic bowel. It is essential to initiate these physiological and safe procedures as soon as possible after sustaining the injury; this will lead to better results and to the elimination of inadequate intestinal maneuvering in the future Spinal Cord (2000) 38, 301 - 308. PMID- 10822404 TI - The effect of frequency and mode of sports activity on the psychological status in tetraplegics and paraplegics. AB - OBJECTIVE: To examine whether the psychological benefits of sports activity differ between tetraplegics and paraplegics with spinal cord injury, and investigate the effect of frequency and modes of sports activity on the psychological benefits. METHODS: The Self-rating Depression Scale (SDS), State Trait Anxiety Inventory (STAI) and Profiles of Mood States (POMS) were administered to 169 male individuals with spinal cord injury (mean age=42.7 years) including 53 tetraplegics and 116 paraplegics. The subjects were divided into four groups according to their frequencies of sports activity; High-active (more than three times a week; n=32), Middle-active (once or twice a week, n=41), Low-active (once to three times a month, n=32), and Inactive (no sports participation, n=64). RESULTS: Analysis of variance revealed significant differences in depression for SDS, trait anxiety for STAI and depression and vigor for POMS among the groups. High-active group showed the lowest scores of depression and trait anxiety and the highest score of vigor among the four groups. In contrast, no significant difference was found for any psychological measurements between tetraplegics and paraplegics. In addition, there was no significant difference for any psychological measurements among modes (wheelchair basketball, wheelchair racing, wheelchair tennis and minor modes). CONCLUSIONS: These findings demonstrated that sports activity can improve the psychological status, irrespective of tetraplegics and paraplegics, and that the psychological benefits are emphasized by sports activity at high frequency. PMID- 10822405 TI - Radiographic evaluation of bioactive glass-ceramic grafts in postero-lateral lumbar fusion. AB - STUDY DESIGN: Follow-up study of bioactive glass-ceramic graft for postero lateral fusion (PLF) in the lumbar spine was performed using plain radiography. OBJECTIVES: To investigate the fusion state of the porous stick type of bioactive glass-ceramic which has been used in PLF in the lumbar spine, by plain radiography, and to evaluate the usefulness of this technique. METHODS: Fourteen patients who underwent PLF using apatite and wollastonite-containing glass ceramic (A-W. GC) were followed-up for more than 1 year and 6 months by plain radiography. RESULTS: Six patients who were followed up for more than 2 years were considered to have obtained subtotal or total fusion. CONCLUSION: PLF using A-W. GC mixed with autologous grafted bone can preserve bone stock without the need to harvest a bone graft from the iliac crest and will be useful for PLF in the lumbar spine. PMID- 10822406 TI - Osteonecrosis after treatment for heterotopic ossification in spinal cord injury with the combination of surgery, irradiation, and an NSAID. AB - STUDY DESIGN: Case report. OBJECTIVE: Heterotopic ossification (HO) is a frequent complication in spinal cord injury (SCI) that is often difficult to treat. Although surgery may become necessary, operative resection has been associated with complications and poor outcome due to a high recurrence rate. Additional methods of treatment to reduce the recurrence rate have been developed, including post operative irradiation and NSAIDs. This article presents three patients, who developed an osteonecrosis of the femoral head after the combined treatment for HO of surgery, irradiation, and an NSAID. PMID- 10822407 TI - Milk of calcium in the inferior calyx of a hydronephrotic kidney in a tetraplegic patient - a diagnosis to be made before scheduling for extracorporeal shock wave lithotripsy. AB - STUDY DESIGN: A Case Report of renal milk of calcium in a tetraplegic subject. OBJECTIVES: To increase the awareness of renal milk of calcium in spinal cord injury (SCI) physicians. Renal milk of calcium contains a colloidal suspension of calcium crystals. Since upright views of the kidneys are not performed in tetraplegic subjects, the renal milk of calcium may be misinterpreted as renal lithiasis by routine radiography taken in supine position. SETTING: Regional Spinal Injuries Centre, Southport, England. METHOD: In a 41-year-old male with traumatic tetraplegia, X-ray of abdomen in supine position showed multiple opacities in the region of the left kidney. These radio opaque shadows were interpreted as renal calculi. Subsequently, computed tomography (CT) of the kidneys was performed. RESULTS: CT confirmed the presence of calculi in the mid polar calyx. However, the density situated in the inferior calyx of the hydronephrotic left kidney exhibited a horizontal upper edge. This specific radiological finding as observed in the CT of kidneys, provided the clue to the presence of milk of calcium in the inferior calyx of the hydronephrotic left kidney. CONCLUSION: As plain film of the abdomen in standing position is not performed in SCI patients, physicians caring for SCI patients should have a high index of suspicion for renal milk of calcium. Prompt diagnosis of renal milk of calcium will help to avoid unnecessary surgery, or extracorporeal shock wave lithotripsy. PMID- 10822408 TI - Slow ascending myelopathy, tetraplegia, carcinoma of the bladder and amyloidosis in a patient with ankylosing spondylitis. AB - OBJECTIVE: We report a case of slow ascending myelopathy in a patient with ankylosing spondylitis (AS). DESIGN: Case report of a 60-year-old patient suffering from AS, who developed over a period of 39 years a slow ascending myelopathy leading to tetraplegia, squamous cell carcinoma of the bladder and amyloidosis of the small intestine secondary to neuropathic bladder and bowel. SETTING: Department and Outpatient's Department of Neurological Rehabilitation Sheba Medical Center, Tel Hashomer, Israel. SUBJECT: Single patient case report. MAIN OUTCOME MEASURE: Clinical follow-up of the patient between the years 1959 - 1998. RESULTS: Physical examination disclosed deteriorating incomplete tetraplegia with hypotonia and hyporreflexia. Neurogenic bladder and bowel complicated to squamous cell carcinoma and amyloidosis. CONCLUSION: To our knowledge, flaccid tetraplegia associated with AS, has never been reported in the literature. The possibility of vascular compression by the ankylosed spine causing the clinical picture of flaccid tetraplegia in this patient is discussed. PMID- 10822409 TI - Foreword PMID- 10822411 TI - Modern evaluation of the hypertensive patient: autonomic tone in cardiovascular disease and the assessment of heart rate variability. AB - Analysis of heart rate variability (HRV) permits an assessment of sympathetic and parasympathetic activity from EKG recordings. Analysis of HRV may be performed in both the time and frequency domain by the application of mathematical principles of signal processing. HRV demonstrates abnormalities in myocardial infarction, sudden death, heart failure, autonomic neuropathy and hypertension. The technique is useful for assessing prognosis and for evaluating therapeutic interventions. PMID- 10822410 TI - Circadian variation of blood pressure and the assessment of antihypertensive therapy. AB - The inherent variability of activity and the awake-sleep cycle creates changes in hemodynamics that may influence the outcome of a variety of cardiovascular disorders. Recently, data suggest that increased variability of blood pressure may promote excessive hypertensive target organ disease. Several epidemiologic studies have demonstrated that myocardial ischemia, myocardial infarction and sudden cardiac death have an excess incidence in the first several hours post awakening. Additionally, surveys of the incidence of stroke (both ischemic and hemorrhagic) have shown an excess for the hours between 8 a.m. and noon. The pathophysiologic bases for the increased number of cardiac and cerebrovascular events in the early morning hours may be both due to hemodynamic and hematorrheologic factors. During the past several years, therapeutic studies have evaluated the effects of antihypertensive therapies on blood pressure and heart rate during the circadian or 24 h period. There are a few studies that have evaluated nighttime dosing of conventional antihypertensive therapies but these have generally been statistically underpowered to demonstrate differences of morning versus evening dosing on circadian blood pressure. Since the timing of dosing (and hence the delivery of antihypertensive therapy) may be clinically relevant, prospective and well-performed clinical trials of chronotherapy are necessary. In this review, we evaluate new data on the clinical impact of cardiovascular chronotherapy and the importance of timing of dosing on pharmacodynamics. PMID- 10822412 TI - One's grandparents as the determinant of effective antihypertensive therapy. AB - We have known for 70 years that familial factors influence blood pressure level. Modern genetics suggest that 30-40% of blood pressure variation reflects genetic factors. We have known for more than 40 years that patients differ in their response to specific antihypertensive agents, with little insight beyond demographics into the responsible mechanism for the variation in that response. Only recently has interest been evident in attempts to bring these two questions together. Specifically, do familial factors influence the response to specific antihypertensive therapy? One example involves the attempt to use the response to angiotensin- converting enzyme (ACE) inhibitors as an intermediate phenotype. An analogous attempt involves recognition of the fact that ACE inhibition reverses many of the features of non-modulation- which is an expression of an angiotensinogen (AGT) gene polymorphism. The ESPRIT Study was the first to enroll hyper-tensive sibling pairs into a therapeutic trial, designed to assess responses to ACE inhibitor treatment with lisinopril. This pathway of investigation is likely to be very fruitful over the next several decades, to the point that 'pharmacogenetics' will be an element in selecting antihypertensive therapy. PMID- 10822413 TI - Left ventricular hypertrophy: a potent cardiovascular risk factor and its relationship to office and ambulatory blood pressure. AB - Left ventricular hypertrophy (LVH) is an important cardiovascular risk factor. The presence of LVH carries risk independent of hypertension. LVH can be detected non-invasively using electrocardiography or echocardiography. Clinical studies have consistently shown that ambulatory blood pressure is a stronger correlate of left ventricular mass than office blood pressure. Furthermore, treatment-induced decreases in left ventricular mass index are also more tightly related to reductions in ambulatory blood pressure than reductions in office blood pressure. The primary intervention for subjects with hypertension and LVH is optimal blood pressure control. Several small studies now suggest that therapeutic changes resulting in regression of left ventricular mass also confer a reduction in cardiovascular risk. Therefore, LVH is a serious negative risk factor that is more closely related ambulatory rather than office blood pressure. Fortunately, current evidence suggests that optimal antihypertensive therapy resulting in regression of hypertrophy will reduce at least short-term cardiovascular events. Physicians need to be more aware of LVH as a cardiovascular risk factor. PMID- 10822414 TI - Obesity-associated hypertension: hypothesized link between etiology and selection of therapy. AB - The association of obesity and hypertension is characterized hemodynamically by an increase in absolute circulating intravascular volume that induces increased cardiac output and total peripheral resistance that remains inappropriately normal. These changes constitute the hemodynamic basis for the increase in blood pressure in obesity. Obesity-associated hypertension is also characterized by an abnormal renal response, including increased renal blood flow and a rise in glomerular and interstitial pressures. These hemodynamic changes induce the following structural changes in the heart: enlarged left atrial, ventricular and aortic root diameters as well as increased posterior septal wall thickness and left ventricular mass. The hemodynamic changes in the kidneys generate higher glomerular volume and increased interstitial infiltrate, which may cause compression of the tubules and blood vessels of the renal medulla. Weight reduction is an effective tool in the control of blood pressure, and significantly reduces the metabolic and hemodynamic derangements that occur with obesity. However, since weight reduction compliance is difficult to maintain, pharmacological agents are often needed to control blood pressure. Angiotensin converting enzyme inhibitors, calcium channel blockers and alpha- adrenergic blocking agents may be the most appropriate therapy for obesity-associated hypertension since they intervene with some of the previously described pathophysiological conditions. PMID- 10822415 TI - Foreword: what have we learned from ambulatory blood pressure monitoring during the past five years? PMID- 10822416 TI - What is the 'normal' 24 h, awake, and asleep blood pressure? AB - The clinical use of ambulatory blood pressure monitoring requires that a 'normal' limit be identified. However, because the relationship between blood pressure and risk is continuous, any such limit is arbitrary. Various different methods have been used to define a normal limit, and can be divided into those that rely solely on blood pressure measurement, and those that relate blood pressure to target-organ damage or risk. First, several population studies of 'normotensive' people have identified the 95th centile of ambulatory blood pressure, with generally consistent findings, but with the methodological flaw that 'hypertensive' individuals must first be excluded. Second, the ambulatory blood pressure that corresponds to a threshold value of clinic blood pressure (e.g. 140/90 mmHg) can be identified from the regression equation linking the two measures of blood pressure. Third, the percentage of readings greater than a threshold value (the 'blood pressure load') can be specified; this method is arbitrary, and makes the unwarranted assumption that there is a threshold value above which blood pressure causes harm. Fourth, the ambulatory blood pressure above which the prevalence of target-organ damage begins to increase significantly can be identified. Fifth, the relationship between ambulatory blood pressure and cardiovascular risk can be examined in prospective studies. Despite these very different approaches, with a few notable exceptions there is reasonable agreement between the methods, the consensus being that, for daytime ambulatory blood pressure, the normal value is about 135/85 mmHg. PMID- 10822417 TI - What are the influences of salt, potassium, the sympathetic nervous system, and the renin-angiotensin system on the circadian variation in blood pressure? AB - Nocturnal blood pressure patterns are expressed as a consequence of both intrinsic circadian rhythms and the quantity and quality of sleep. A range of neurohumoral factors have been either inferred or definitively proven to influence the circadian blood pressure pattern. In this regard, changes in atrial natriuretic peptide, plasma renin activity and plasma aldosterone have been evaluated as to the influence each might have in determining diurnal blood pressure patterns. As an example, a clear pattern of nocturnal increase in plasma renin activity has been observed; however, the relationship between this change and nocturnal blood pressure remains vague. In contrast, change in sympathetic nervous system activity has most commonly been associated with the conversion of a pattern of nocturnal dips in blood pressure to one of non-dipping. Increasingly, dietary intake patterns, characterized by high sodium or low potassium content, or both, are recognized as modifiers of the normal decrease in nocturnal blood pressure, again in favor of producing a non-dipping pattern. The impact of these nutritional patterns on nocturnal blood pressure change is recognized to be most prominent in salt-sensitive individuals. Modifications of dietary sodium intake, such that it is decreased or potassium intake is increased, or both, are now recognized as means by which a nocturnal non-dipping pattern can be converted to a dipping pattern. Additional studies of an integrative nature will be necessary to obtain more complete definition of the dynamic interplay between nutrition and various neurohumoral axes in determining how nocturnal blood pressure patterns are expressed. PMID- 10822418 TI - How well does ambulatory blood pressure predict target-organ disease and clinical outcome in patients with hypertension? AB - For many years, it has been evident that ambulatory blood pressure monitoring is superior to the measurement of office blood pressure as a predictor of target organ involvement in patients with hypertension. Until recently, there were far fewer data on the relationship between 24 h ambulatory blood pressure and cardiovascular outcomes such as myocardial infarction, stroke, and cardiovascular death. In 1983, Perloff et al. published their seminal report on awake ambulatory blood pressure as a predictor of cardiovascular outcomes. During the 16 years that have passed since that publication, several additional prospective ambulatory blood pressure studies have been completed, in five different countries. The basis for all these investigations has been to assess the predictive value of ambulatory blood pressure as a determinant of either cardiovascular morbidity (myocardial infarction, cerebrovascular accidents, and vascular surgical procedures) or mortality. With the exception of the Systolic Hypertension in Europe (Syst-Eur) trial, all these studies have been uncontrolled for therapeutic interventions. Typically, the average follow-up period for each trial has been 3-9 years. All these studies have shown that ambulatory blood pressure is a much better predictor of cardiovascular events than the standard office or clinic pressure. In addition, hypertensive patients whose nocturnal (or sleep) blood pressure remains high (that is, those who have a 'non-dipper' circadian blood pressure profile) have a worse outcome than patients whose nocturnal blood pressure decline is at least 10%. These data all support the desirability of increased utilization of 24 h ambulatory blood pressure monitoring in clinical trials of antihypertensive drugs and in the management of hypertensive patients in clinical practice. PMID- 10822419 TI - What are the approaches for evaluating antihypertensive treatment by 24 h ambulatory blood pressure monitoring? AB - Measurements of trough blood pressure in a clinic setting have been the traditional method of assessing the efficacy of antihypertensive agents. The duration of action of antihypertensive drugs has been assessed by calculation of a trough-to-peak ratio; drugs with a trough-to-peak ratio greater than 50% are typically given once-a-day indications. However, the use of clinical measurements to assess antihypertensive agents can be misleading. Ambulatory blood pressure monitoring is a simple technique that provides accurate and reproducible data on both the efficacy and duration of action of antihypertensive agents. Although several complicated techniques have been used for the analysis of ambulatory blood pressure data, studies have demonstrated that calculation of simple blood pressure means (24 h mean, day-time mean and night-time mean) will provide all the data required to assess the efficacy of a drug. Calculations of systolic and diastolic load also provide useful information, and the index correlates closely with target-organ damage. Assessing the reduction of blood pressure during the last 2-6 h of the dosing interval provides critical information on the duration of action of agents with once-a-day dosing. Trough-to-peak ratio can also be calculated from an ambulatory blood pressure monitor. Furthermore, a simple line graph constructed from hourly means makes available, at a simple glance, a large amount of information about a drug. The reproducibility of ambulatory monitoring, together with the absence of placebo effect and the ability to exclude patients with white-coat hypertension, make the technique an extremely powerful tool for the assessment of antihypertensive agents that clearly provides more data on the efficacy and duration of action of an antihypertensive agent than do traditional clinical measurements. PMID- 10822421 TI - "The scarlet E": epilepsy is still a burden. PMID- 10822422 TI - Response to L-dopa in PD: the long and the short of it. PMID- 10822423 TI - Practice parameter: anticonvulsant prophylaxis in patients with newly diagnosed brain tumors. Report of the Quality Standards Subcommittee of the American Academy of Neurology. PMID- 10822425 TI - Access www.neurology.org now for full-text articles PMID- 10822424 TI - "The scarlet E": the presentation of epilepsy in the English language print media. AB - OBJECTIVE: To examine the coverage of epilepsy in English language newspapers and magazines to determine how they portray the medical risks associated with epilepsy, whether they report research and treatment advances accurately, whether stigmatizing biases toward persons with epilepsy persist, and to examine the sources of errors in reporting about epilepsy. BACKGROUND: The print media reflect and shape current views about epilepsy and other neurologic conditions. They also have the potential to further misconceptions about neurologic issues and particular brain disorders. Persistent myths about epilepsy, such as the ancient belief that it is a demonic disorder, can result in discrimination, emotional difficulties, and reluctance to seek effective treatment. METHODS: A large commercial database was used to search for stories about epilepsy from approximately 2,000 English language newspapers and popular magazines. Two epileptologists independently classified story themes and main sources, and screened for the presence of gross errors in 210 stories about epilepsy or seizures. The authors analyzed the metaphors and terminology used to describe seizures and epilepsy. RESULTS: The majority of English language print stories about epilepsy were accurate depictions of social and medical issues regarding the disorder, most commonly depictions of persons overcoming epilepsy and announcements of new therapies and reports of scientific advances. Thirty-one percent of the stories, however, contained gross errors, most commonly scientific inaccuracy, exaggerated treatment claims, and overestimates of the risks of dying during a seizure. New drug therapies were often described inaccurately by physicians and pharmaceutical spokespersons as curative and without side effects. Patients and their families frequently overemphasized the risk of dying during a seizure and misstated medical issues. Most celebrities with recurring seizures denied having epilepsy. Seizures were described with demonic imagery in 6% of stories. United States epilepsy associations discourage labeling patients as "epileptics"; however, the term was used in 45% of stories. CONCLUSION: Physicians and reporters should be aware of both professional and popular biases that influence the print media's presentation of the causes and consequences of epilepsy. PMID- 10822426 TI - Stroke prevention: narrowing the evidence-practice gap. AB - Many interventions reduce stroke risk. However, the full benefits of these interventions are not realized at current levels of utilization, as nearly all evidence-based or guideline-endorsed stroke prevention services are underused. The cause for such underuse is multifactorial and includes factors relating to both patients and providers, as well as to a health care system that has de emphasized prevention at the expense of acute, technologically based care. Much like the evidence for stroke interventions themselves, there is a growing literature to support methods of implementing research evidence into clinical practice. There is still much to learn, however, about the effectiveness of interventions aimed at achieving changes in stroke prevention practice or the delivery of stroke prevention care. Nevertheless, there are many opportunities for providers, managed care organizations, and government to close the evidence practice gap that exists for stroke prevention services. These opportunities exist in both the inpatient and outpatient setting, and depend on the neurologist taking a leading role in emphasizing the critical importance of risk factor identification and modification in all patients at risk for stroke. PMID- 10822427 TI - Platelet APP isoform ratios correlate with declining cognition in AD. AB - BACKGROUND: Platelets and neurons both contain large quantities of two carboxyl truncated 120 to 130 and 110 kDa Alzheimer amyloid precursor proteins (APPs). Platelets taken from patients with AD have been reported to contain a reduced ratio of these APPs. OBJECTIVE: To further study the AD specificity of reduced platelet APP ratios and to determine whether, after 3 years, cognitive losses in AD are accompanied by similarly reduced platelet APP ratios. METHODS: To test the AD specificity of reduced platelet APP ratios, we quantitated these APPs in eight patients with PD and six patients with hemorrhagic stroke (HS). To determine whether further cognitive losses correlate with platelet APP ratio reductions in patients with AD, the authors re-examined platelet APPs and Mini-Mental State Examination (MMSE) scores of 10 patients with AD and 11 controls, who were tested 3 years ago. APP ratios were determined by the average of six assays using Western blotting with m22C11 monoclonal antibody, enhanced chemoluminescence, and digital scanning of autoradiographs. RESULTS: APP ratios were normal in the patients with PD and HS, further supporting the AD specificity of this assay. After 3 years, the MMSE scores and APP ratios of our control subjects changed by <4%. However, the average MMSE scores of our patients with AD declined from 16.4 to 8.3, and their average 120 to 130/110 kDa APP ratios declined from 5.8 to 3.6. The difference between AD and control APP ratios, with no overlap, is significant and the correlation between the 3-year decline in AD MMSE scores and reduced APP ratios (r = 0.69) was significant. CONCLUSIONS: Although the number of subjects analyzed was limited, reduced platelet APP ratios appear to be a specific biological marker of AD and a biological index of the severity of cognitive loss in AD. PMID- 10822428 TI - The long-duration response to L-dopa in the treatment of early PD. AB - OBJECTIVE: To investigate the long-duration response (LDR) to L-dopa resulting from different regimens of L-dopa. BACKGROUND: In clinical practice, L-dopa is usually administered without considering the LDR due to the drug. Moreover, it has not been established whether in early PD a multiple daily intake of small doses of L-dopa may induce a sustained LDR. METHODS: Twenty-four patients with early PD underwent a double-blind, crossover trial, comparing three different 15 day treatment periods with L-dopa: treatment A (250 mg every 24 hours); treatment B (250 mg every 8 hours); and treatment C (125 mg every 8 hours). After completion, 20 patients underwent a subsequent open-label randomized trial with prolonged treatments (250 mg every 24 hours or 125 mg every 8 hours) up to 3 months. LDR was measured at the end of each treatment. RESULTS: All patients achieved a sustained LDR after treatments A and B, whereas only 17% of patients reached a sustained LDR after treatment C. Overall, the LDRs resulting from treatments A and B had similar magnitude and were larger than the LDR deriving from treatment C. After 3 months of prolonged treatments, only three of 10 patients treated with 125 mg every 8 hours increased their LDR, whereas all 10 patients treated with 250 mg every 24 hours had a maximal and stable LDR. CONCLUSIONS: Sustained LDR to L-dopa is dependent on the amount of the single doses of the drug. A regimen scheduling small, divided doses during the day, as done in clinical practice, is a questionable therapy for the achievement of a sustained LDR. PMID- 10822429 TI - Alpha-synuclein cortical Lewy bodies correlate with dementia in Parkinson's disease. AB - BACKGROUND: Dementia is a frequent complication of idiopathic parkinsonism or PD, usually occurring later in the protracted course of the illness. The primary site of neuropathologic change in PD is the substantia nigra, but the neuropathologic and molecular basis of dementia in PD is less clear. Although Alzheimer's pathology has been a frequent finding, recent advances in immunostaining of alpha synuclein have suggested the possible importance of cortical Lewy bodies (CLBs) in the brains of demented patients with PD. METHODS: The brains of 22 demented and 20 nondemented patients with a clinical and neuropathologic diagnosis of PD were evaluated with standard neuropathologic techniques. In addition, CLBs and dystrophic neurites were identified immunohistochemically with antibodies specific for alpha-synuclein and ubiquitin; plaques and tangles were identified by staining with thioflavine S. Associations between dementia status and pathologic markers were tested with logistic regression. RESULTS: CLBs positive for alpha-synuclein are highly sensitive (91%) and specific (90%) neuropathologic markers of dementia in PD and slightly more sensitive than ubiquitin-positive CLBs. They are better indicators of dementia than neurofibrillary tangles, amyloid plaques, or dystrophic neurites. CONCLUSION: CLBs detected by alpha synuclein antibodies in patients with PD are a more sensitive and specific correlate of dementia than the presence of Alzheimer's pathology, which was present in a minority of the cases in this series. PMID- 10822430 TI - Cervical dural sac and spinal cord in juvenile muscular atrophy of distal upper extremity. AB - OBJECTIVE: To investigate specificity and significance of dynamic changes of the cervical dural sac and spinal cord during neck flexion in juvenile muscular atrophy of the distal upper extremity. BACKGROUND: The disorder affects young people-predominantly men-and is progressive for several years. One autopsy case showed ischemic necrosis of the cervical anterior horn, suggesting that the disorder is a type of cervical myelopathy. Some authors classify it as monomelic amyotrophy, implying that it is a focal motor neuron disease. METHODS: Neuroradiologic examinations including myelography, CT myelography, and MRI in a fully flexed neck position were performed on 73 patients with this disorder and on 20 disease control subjects. RESULTS: A distinctive finding in the disorder was forward displacement of the cervical dural sac and compressive flattening of the lower cervical cord during neck flexion. The forward displacement was significantly greater in patients with disease duration less than 10 years than in age-matched control subjects and patients in a late, nonprogressive stage. CONCLUSIONS: Radiologic abnormalities of the lower cervical dural sac and spinal cord support the hypothesis that this disorder is a type of cervical myelopathy. PMID- 10822431 TI - FSH dystrophy 4q35 deletion in patients presenting with facial-sparing scapular myopathy. AB - OBJECTIVE: To evaluate the incidence of the facioscapulohumeral dystrophy (FSHD) 4q35 deletion in patients with facial-sparing scapular myopathy. BACKGROUND: Scapular winging is typical of FSHD but may also be prominent in other muscle disorders including scapuloperoneal syndromes. With DNA testing, it is possible to determine if patients with facial-sparing scapular myopathy have FSHD. METHODS: Fourteen of 17 unrelated patients with facial-sparing scapular myopathy, seen over a 7-year period at a regional neuromuscular center, agreed to have DNA testing for FSHD. The clinical and laboratory features of these patients were also noted. RESULTS: Of the 14 patients, 10 (71%) had restriction fragments consistent with the 4q35 deletion. The mean size of the smaller fragment following EcoRI digestion was 29.5 kb (range 20 to 39). The mean age at onset was 19.9 years; at presentation, 44.7 years. Except for the absence of facial weakness, most patients had clinical and laboratory features otherwise consistent with FSHD. Five patients (50%) had a positive family history of similar weakness. Following removal of outliers, the Pearson correlation coefficient (r) value between EcoRI fragment size and age at onset was 0.64, and between fragment size and limb muscle strength, 0.64. CONCLUSION: The FSHD 4q35 deletion was found in 71% of the facial-sparing scapular myopathy patients. They otherwise resemble typical FSHD patients in age at onset, physical characteristics, and association between fragment size and disease severity. PMID- 10822432 TI - Peripheral and central conduction abnormalities in diabetes mellitus. AB - OBJECTIVES: To investigate peripheral and central somatosensory conduction in patients with diabetes. METHODS: The authors recorded sensory nerve action potentials and 5-channel somatosensory evoked potentials (SEPs) with noncephalic reference after median nerve stimulation in 55 patients with diabetes and 41 age- and height-matched normal subjects. The authors determined onset or peak latencies of the Erb's potential (N9) and the spinal N13-P13 and the cortical N20 P20 components, and obtained the central conduction time (CCT) by onset-to-onset and peak-to-peak measurements. RESULTS: Both onset and peak latencies of all SEP components were prolonged in patients with diabetes. The mean onset CCT in the diabetic group was 6.3 +/- 0.5 msec (mean +/- SD)-significantly longer than that in the control group (6.1 +/- 0.2 msec)-whereas no significant difference was found in the peak CCT. The amplitudes of N9 and N13-P13 components (but not N20 P20) were significantly smaller in the diabetic group. The peripheral sensory conduction velocity was also decreased in the diabetic group, but there was no significant correlation between peripheral conduction slowing and the onset of CCT prolongation. CONCLUSIONS: Diabetes affects conductive function in the central as well as peripheral somatosensory pathways. The CCT abnormality does not coincide with lowering of the peripheral sensory conduction. The current results do not favor a hypothesis that a central-peripheral distal axonopathy plays an important role in development of diabetic polyneuropathy. PMID- 10822433 TI - A novel approach to stroke rehabilitation: robot-aided sensorimotor stimulation. AB - OBJECTIVE: In patients with stroke, the authors tested whether additional sensorimotor training of the paralyzed or paretic upper limb delivered by a robotic device enhanced motor outcome. METHODS: Fifty-six patients with stroke and hemiparesis or hemiplegia received standard poststroke multidisciplinary rehabilitation, and were randomly assigned either to receive robotic training (at least 25 hours) or exposure to the robotic device without training. Outcomes were assessed by the same masked raters, before treatment began and at the end of treatment, with the upper extremity component of the Fugl-Meyer Motor Assessment, the Motor Status score, the Motor Power score, and Functional Independence Measurement. RESULT: The robot treatment and control group had comparable clinical characteristics, lesion size, and pretreatment impairment scores. By the end of treatment, the robot-trained group demonstrated improvement in motor outcome for the trained shoulder and elbow (Motor Power score, p < 0.001; Motor Status score, p < 0. 01) that did not generalize to untrained wrist and hand. The robot-treated group also demonstrated significantly improved functional outcome (Functional Independence Measurement-Motor, p < 0. 01). CONCLUSION: Robot delivered quantitative and reproducible sensorimotor training enhanced the motor performance of the exercised shoulder and elbow. The robot-treated group also demonstrated improved functional outcome. When added to standard multidisciplinary rehabilitation, robotics provides novel therapeutic strategies that focus on impairment reduction and improved motor performance. PMID- 10822434 TI - Better outcome after stroke with higher serum cholesterol levels. AB - OBJECTIVE: To examine whether serum cholesterol levels have any prognostic value in the first month following acute ischemic stroke. BACKGROUND: Although the association between serum cholesterol levels and cerebrovascular disorders has been extensively studied, the relationship between cholesterol levels and outcome following ischemic stroke has not been investigated. METHODS: Using data from 3,273 consecutive patients with first-ever ischemic stroke, the authors compared poor functional outcome (severe disability or death) at 1 month in patients with high cholesterol (total serum cholesterol greater than 6.5 mmol/L or 250 mg/dL) and normal cholesterol (level equal to or less than 6.5 mmol/L or 250 mg/dL). Data were analyzed by univariate and multivariate analysis. RESULTS: In comparison with patients with normal cholesterol levels, patients with high cholesterol levels had a 2.2-fold lower risk of death (p = 0.002) and a 2.1-fold lower risk of poor functional outcome at 1 month (p < 0.001). After adjustment for known confounding variables, multivariate analysis showed that higher cholesterol levels remained an independent predictor of better functional outcome (OR 0.48, CI 0. 34 to 0.69, p < 0.001). CONCLUSIONS: The authors' findings suggest that higher levels of cholesterol are associated with a better outcome in the early phase after ischemic stroke. PMID- 10822435 TI - Estrogen use, APOE, and cognitive decline: evidence of gene-environment interaction. AB - OBJECTIVE: APOE-epsilon4 increases the risk of cognitive decline, while elderly women who take estrogen may have less risk of cognitive decline. The authors sought to determine whether estrogen use modifies the association between APOE epsilon4 and cognitive decline. METHOD: - As part of the Cardiovascular Health Study, 3,393 Medicare-eligible women (> or =65 years) were randomly selected and recruited from Sacramento County, CA; Washington County, MD; Forsyth County, NC; and Pittsburgh, PA. Cognitive testing was administered annually; the authors studied the 2,716 women with cognitive testing on > or =2 visits. They analyzed change in score on the Modified Mini-Mental State Examination (3MS) as a function of estrogen use, APOE genotype, and baseline common and internal carotid artery wall thickening. RESULTS: A total of 297 (11%) women were current estrogen users and 336 (12%) were past estrogen users. Over the 6-year average follow-up, baseline current users declined 1.5 points on the 3MS whereas never users declined 2.7 points (p = 0.023). Compared with epsilon4-negative women, epsilon4 positive women had a greater adjusted hazard ratio of cognitive impairment (3MS < 80), hazard risk [HR] = 1.47; 95% CI, 1.13 to 1.90. There was an interaction between estrogen use and epsilon4 presence (p = 0.037). Among epsilon4-negative women, current estrogen use reduced the risk of adjusted cognitive impairment compared with never users by almost half (HR = 0.59; 95% CI, 0.36 to 0.99), whereas, it did not reduce the risk among epsilon4-positive women (current use, HR = 1.33; 95% CI, 0.74 to 2.42). Compared with never use, current estrogen use was associated with less internal and common carotid wall thickening in epsilon4 negative women but not in epsilon4-positive women (p for interaction < 0.05 for both). Differences remained after adjusting for age, education, race, and stroke. CONCLUSIONS: Estrogen use was associated with less cognitive decline among epsilon4-negative women but not epsilon4-positive women. Potential mechanisms, including carotid atherosclerosis, by which epsilon4 may interact with estrogen and cognition warrant further investigation. PMID- 10822436 TI - Visual attention impairments in Alzheimer's disease. AB - BACKGROUND: Impaired attention can hinder information processing at multiple levels and may explain aspects of functional decline in aging and dementia. Impairments of attention in early AD may contribute to performance reductions in other cognitive domains, including memory and executive functions. METHOD: - The authors analyzed the scores on a battery of tests of attention and cognitive abilities in 64 older individuals: 42 with mild AD and 22 control subjects without dementia. The authors tested the hypotheses that patients with AD would have impairments of visual attention, and that these impairments would correlate with dysfunction in other key cognitive domains. RESULTS: Patients with AD performed significantly worse than control subjects on measures of sustained attention, divided attention, selective attention, and visual processing speed. The differences were not due to differences in age, education, or basic visual function. Strong relationships were identified between reduced attention skills and overall cognitive impairment. CONCLUSIONS: Deterioration of attention abilities occurs in early stages of AD, and likely contributes to functional decline in these patients. More routine assessment of visual attention deficits could give a more accurate measure of functionally useful perception in patients with AD who show normal visual acuity and visual fields, perhaps providing useful clues to diagnosis and staging. PMID- 10822437 TI - Cytoskeleton proteins in CSF distinguish frontotemporal dementia from AD. AB - OBJECTIVE AND BACKGROUND: To investigate the CSF levels of tau and the light neurofilament protein (NFL) in patients with frontotemporal dementia (FTD) and other common dementia disorders as well as normal control subjects. Both proteins have been implicated in the pathophysiology of FTD. METHODS: CSF levels of tau and NFL were investigated in 18 patients with FTD, 21 patients with early-onset AD (EAD), 21 patients with late-onset AD (LAD), and 18 age-matched control subjects. RESULTS: Mean +/- SD CSF NFL levels were increased in patients with FTD (1442 +/- 1183 pg/mL; p < 0.05) and LAD (1006 +/- 727 pg/mL; p < 0.001) compared with control subjects (241 +/- 166 pg/mL) and in LAD compared with EAD (498 +/- 236 pg/mL; p < 0.05), and tended to be increased in FTD compared with EAD. CSF tau levels were increased in EAD (751 +/- 394 pg/mL; p < 0.01) and LAD (699 +/- 319 pg/mL; p < 0.01) compared with control subjects (375 +/- 170 pg/mL), and in EAD (p < 0.001) and LAD (p < 0. 01) compared with FTD (354 +/- 140 pg/mL). CSF NFL correlated positively with degree of cognitive impairment in FTD (r = 0.59; p < 0.05) and LAD (r = 0.61; p < 0.01). No significant differences were found in CSF NFL or CSF tau when comparing patients who did and did not possess the APOE epsilon4 allele within each diagnostic group. CONCLUSION: The results suggest a differential involvement of these cytoskeleton proteins in FTD and EAD, with NFL primarily involved in the pathophysiology of FTD and tau in that of EAD. The increase in CSF NFL found in LAD might reflect the white-matter degeneration found in a proportion of LAD cases. PMID- 10822438 TI - Incidence of and risk factors for hallucinations and delusions in patients with probable AD. AB - OBJECTIVE: To examine the incidence of and risk factors for hallucinations and delusions associated with patients clinically diagnosed with probable AD. BACKGROUND: Estimates of the incidence of psychosis in AD range widely from 10% to 75%. The risk factors for psychosis of AD are not known, although multiple studies indicate that AD patients with psychosis demonstrate greater cognitive and functional impairment. METHODS: The authors conducted psychiatric evaluations of 329 patients with probable AD from the University of California at San Diego Alzheimer's Disease Research Center to determine the incidence of hallucinations and delusions. They examined data from annual clinical and neuropsychological evaluations to determine whether there were specific risk factors for the development of hallucinations and delusions. RESULTS: Using Cox survival analyses, the cumulative incidence of hallucinations and delusions was 20.1% at 1 year, 36.1% at 2, 49.5% at 3, and 51.3% at 4 years. Parkinsonian gait, bradyphrenia, exaggerated general cognitive decline, and exaggerated semantic memory decline were significant predictors. Age, education, and gender were not significant predictors. CONCLUSIONS: The authors found a relatively high incidence of hallucinations and delusions in patients diagnosed with probable AD and suggest that specific neurologic signs, cognitive abilities, and accelerated decline may be predictive markers for their occurrence. PMID- 10822439 TI - A gene on SCA4 locus causes dominantly inherited pure cerebellar ataxia. AB - BACKGROUND: Several different genes or their loci have been identified for autosomal dominant cerebellar ataxia (ADCA). However, other types of ataxia remain unassigned. OBJECTIVE: To identify a new locus for ADCA. METHODS: Six Japanese families with ADCA with pure cerebellar syndrome (ADCA type III) were examined. These families had been molecularly excluded for spinocerebellar ataxia (SCA) types 1 through 3, 5 through 8, and 10. Clinical examination was undertaken, and a genome-wide linkage search was performed on 250 microsatellite DNA markers. RESULTS: Strong evidence for linkage was found with markers on human chromosome 16q, and haplotype and multipoint analyses further refined the gene locus in a 10.9-cM interval between D16S3089 and D16S515. Linkage disequilibrium was further found with the marker D16S3107 within the interval. The locus was exactly the candidate interval of SCA4, a rare form of ADCA clinically characterized by ataxia with sensory neuropathy and pyramidal tract signs. This would suggest that SCA4 and our ADCA type III are likely to be allelic disorders with different clinical features. CONCLUSION: The current study provides evidence that a gene on the SCA4 locus causes a pure cerebellar syndrome. PMID- 10822440 TI - Multimodality imaging for improved detection of epileptogenic foci in tuberous sclerosis complex. AB - OBJECTIVE: Using interictal alpha-[11C]methyl-l-tryptophan ([11C]AMT) PET scan, the authors have undertaken a quantitative analysis of all tubers visible on MRI or 2-deoxy-2-[18F]fluoro-d-glucose ([18F]FDG) PET, to determine the relationship between [11C]AMT uptake and epileptic activity on EEG. BACKGROUND: Tuberous sclerosis complex (TSC) is an autosomal dominant disorder, often associated with cortical tubers and intractable epilepsy. The authors have shown previously that [11C]AMT PET scans show high tracer uptake in some epileptogenic tubers and low uptake in the remaining tubers. METHODS: Eighteen children, age 7 months to 16 years, were studied. Patients underwent video-EEG monitoring, PET scans of [11C]AMT and [18F]FDG, and T2-weighted or fluid-attenuated inversion recovery (FLAIR) MRI. [11C]AMT uptake values were measured in 258 cortical tubers delineated with coregistered MRI or [18F]FDG scans. Uptake ratios were calculated between the [11C]AMT uptake in tubers and those for normal cortex (tuber/normal cortex). Using the region of epileptiform activity, the authors performed receiver operator characteristics (ROC) analysis and determined the optimal uptake ratio for detecting presumed epileptogenic tubers. RESULTS: Tuber uptake ratios ranged from 0.6 to 2.0. Tuber uptake ratios in the epileptic lobes were higher than those in the nonepileptic lobes (p < 0.0001). All 15 patients with focal seizure activity showed one or more lesions with uptake ratio above 0.98 in the epileptic lobe. ROC analysis showed that a tuber uptake ratio of 0.98 resulted in a specificity of 0.91. CONCLUSIONS: Cortical tubers with [11C]AMT uptake greater than or equal to normal cortex are significantly related to epileptiform activity in that lobe. Together, interictal [11C]AMT PET and FLAIR MRI improve the detection of potentially epileptogenic tubers in patients with TSC being evaluated for epilepsy surgery. PMID- 10822441 TI - A hypothetical scheme for the brainstem control of vertical gaze. AB - OBJECTIVES: To develop a hypothetical scheme to account for clinical disorders of vertical gaze based on recent insights gained from experimental studies. METHODS: The authors critically reviewed reports of anatomy, physiology, and effects of pharmacologic inactivation of midbrain nuclei. RESULTS: Vertical saccades are generated by burst neurons lying in the rostral interstitial nucleus of the medial longitudinal fasciculus (riMLF). Each burst neuron projects to motoneurons in a manner such that the eyes are tightly coordinated (yoked) during vertical saccades. Saccadic innervation from riMLF is unilateral to depressor muscles but bilateral to elevator muscles, with axons crossing within the oculomotor nucleus. Thus, riMLF lesions cause conjugate saccadic palsies that are usually either complete or selectively downward. Each riMLF contains burst neurons for both up and down saccades, but only for ipsilateral torsional saccades. Therefore, unilateral riMLF lesions can be detected at the bedside if torsional quick phases are absent during ipsidirectional head rotations in roll. The interstitial nucleus of Cajal (INC) is important for holding the eye in eccentric gaze after a vertical saccade and coordinating eye-head movements in roll. Bilateral INC lesions limit the range of vertical gaze. The posterior commissure (PC) is the route by which INC projects to ocular motoneurons. Inactivation of PC causes vertical gaze-evoked nystagmus, but destructive lesions cause a more profound defect of vertical gaze, probably due to involvement of the nucleus of the PC. Vestibular signals originating from each of the vertical labyrinthine canals ascend to the midbrain through several distinct pathways; normal vestibular function is best tested by rotating the patient's head in the planes of these canals. CONCLUSIONS: Predictions of a current scheme to account for vertical gaze palsy can be tested at the bedside with systematic examination of each functional class of eye movements. PMID- 10822442 TI - Self-injection ictal SPECT during partial seizures. AB - The authors compared ictal SPECT injection performed by medical personnel with self-injection ictal SPECT in six patients with refractory temporal lobe epilepsy. Self-injection was safe and started faster. Self-injection subtraction ictal SPECT coregistered to MRI (SISCOM) was localizing in three patients who had a complex partial seizure, but only one of three patients who had a simple partial seizure, which may limit its usefulness in clinical practice. The localizing information of self-injection was better in three patients, and obviated the need for depth-EEG studies in one patient. PMID- 10822443 TI - Selective loss of vergence control secondary to bilateral paramedian thalamic infarction. AB - The supranuclear pathways for vergence eye movements are poorly understood. The authors report a 57-year-old patient who presented with selective loss of vergence control and dissociation of light and near reaction. MRI showed a symmetric paramedian thalamic infarction without midbrain lesion. The findings suggest that this syndrome is due to an interruption of supranuclear fibers to midbrain vergence neurons. PMID- 10822444 TI - Comparison of stroke hospitalization rates among Mexican-Americans and non Hispanic whites. AB - The authors performed a prospective, community-based pilot stroke surveillance project in Nueces County, TX. Mexican-Americans showed a trend toward higher completed ischemic stroke hospitalization rates compared with non-Hispanic whites. Mexican-Americans were more commonly uninsured (p = 0.007) and were less likely to receive neuroimaging (p = 0.001). Additional studies are needed to confirm this finding and to determine the role of stroke risk factors and access to care variables. PMID- 10822445 TI - Basilar artery vasospasm in postpartum cerebral angiopathy. AB - The reason cerebral edema in postpartum cerebral angiopathy (PPCA) occurs preferentially in the posterior brain is poorly understood. The authors present two patients with PPCA who showed vasospasm occurring earlier and more severely in the basilar artery than in the middle cerebral artery. Our patients demonstrate the difference in vascular change between the anterior and posterior cerebral vessels, explaining the susceptibility of the posterior brain to PPCA. PMID- 10822446 TI - Variable expression of familial Alzheimer disease associated with presenilin 2 mutation M239I. AB - In a family with autopsy-confirmed Alzheimer disease, the authors found a mutation in the presenilin 2 (PS2) gene (PSEN2) that predicts a methionine-to isoleucine change at PS2 residue 239 (M239I), at which a change to valine was known in another family. Phenotypic expression of M239I was highly variable, with disease onset between age 44 and 58 years, and two nonaffected mutation carriers at age 58 and 68 years. The data showed no influence of APOE but were compatible with other possible genetic modifiers of the phenotype or penetrance of M239I. PMID- 10822447 TI - Increased CSF levels of nerve growth factor in patients with Alzheimer's disease. AB - The authors quantitated CSF levels of nerve growth factor (NGF) in patients with AD, nondemented control subjects (CTR), and age-matched patients with major depression (DE). CSF levels of NGF were markedly higher in the AD group than in both the CTR and DE groups (p < 0.01 and p < 0.001). Increased CSF levels of NGF in AD patients may reflect reported accumulation of NGF in the AD brain and may constitute a candidate marker for clinical diagnosis and therapeutic monitoring. PMID- 10822448 TI - Spinal neurenteric cyst presenting in infancy with chronic fever and acute myelopathy. AB - The authors describe the clinical, radiologic, and pathologic features of a neonatal spinal neurenteric cyst (NC) presenting with long-lasting fever and acute myelopathy, and compare this observation with other infants reported in the literature. This observation shows that NC must be considered in the differential diagnosis of acute myelopathy with persistent fever in infancy. Fever is attributed to degenerative changes in the NC, triggering inflammatory cell infiltration and tumor necrosis factor alpha secretion. PMID- 10822449 TI - Activate your online subscription PMID- 10822450 TI - Posterior semicircular canal nystagmus is conjugate and its axis is parallel to that of the canal. AB - A patient with a postoperative fistula of the left posterior semicircular canal is presented. Negative pressure in the external ear canal produced upbeat torsional nystagmus, which was recorded in three dimensions using binocular scleral search coils. The nystagmus was conjugate, without skew deviation, and its trajectory corresponded to the anatomic axis of the left posterior canal. The current study helps validate Ewald's first law in humans: the axis of nystagmus should match the anatomic axis of the semicircular canal that generated it. This law is clinically useful in diagnosing pathology of the vestibular end-organ, such as benign paroxysmal positional vertigo or the superior semicircular canal dehiscence syndrome. PMID- 10822451 TI - Serum S100B levels in patients with HTLV-I-associated myelopathy/tropical spastic paraparesis. PMID- 10822452 TI - Myoclonus secondary to albuterol (salbutamol) instillation. PMID- 10822453 TI - Multifocal brain MRI hypointensities secondary to cardiac catheterization. PMID- 10822454 TI - Hypersexuality in patients with dementia: possible response to cimetidine. PMID- 10822455 TI - Separate representations of static and dynamic touch in human somatosensory thalamus. PMID- 10822456 TI - Neuroimages. Cardiac arrest during partial seizure. PMID- 10822457 TI - Reassessment: vagus nerve stimulation for epilepsy. PMID- 10822458 TI - Brief antiepileptic drug withdrawal prolongs interval to next seizure. PMID- 10822459 TI - Leukoencephalopathy and raised brain lactate from heroin vapor inhalation. PMID- 10822460 TI - A mutation in the microtubule-associated protein tau in pallido-nigro-luysian degeneration. PMID- 10822461 TI - Isolated intracranial hypertension as the only sign of cerebral venous thrombosis. PMID- 10822462 TI - Biology and therapy of epidemic Kaposi's sarcoma. AB - Kaposi's sarcoma, once a rarely seen neoplasm in the West, now occurs in an epidemic fashion in association with acquired immune deficiency syndrome (AIDS). The pathogenesis of Kaposi's sarcoma is still unclear but it appears to be an endothelial neoplasm. Its clinical presentation may be quite subtle and varied. The natural history of Kaposi's sarcoma is still not fully defined, and its rate of progression may be either relatively indolent or aggressive. Therapies include local radiation, recombinant interferon alfa-2a, and cytotoxic chemotherapy. For a subset of patients with Kaposi's sarcoma who were treated with recombinant interferon alfa-2a, the disease is in complete remission, without opportunistic infection, and they appear to be culture-negative for the etiologic retrovirus that causes their immune deficiency. Interferon alfa-2a appears to have antineoplastic efficacy, (and may have antiretroviral efficacy as well) in this epidemic neoplasm. PMID- 10822463 TI - Three consecutive phase II studies of recombinant interferon alfa-2a in advanced malignant melanoma. Updated analyses. AB - In three consecutive Phase II trials of recombinant interferon alfa-2a (rIFN alfa 2a; Roferon-A Hoffmann-La Roche, Nutley, NJ) involving 96 patients with advanced malignant melanoma, an overall response rate of 22% was observed. For all study participants, the median time to disease progression was 1.7 months, and the median survival was six months. Most regressions occurred within one month of commencing therapy, were usually limited to soft tissue metastases, and were transient. However, responses in three patients were long term, lasting 32+, 36+, and 41+ months. A thrice weekly intramuscular dose of 50 x 10(6) U/m2 produced an intolerable flulike illness concomitant with a median weight loss of 5.6 kg. The addition of cimetidine to the same dose in 35 patients was of no therapeutic value. A dose of 12 x 10(6) U/m2 produced clinically acceptable toxicities, and a median weight loss of 2.1 kg. There was no apparent dose response relationship, nor were there any obvious sequelae from antibody formation to interferon alfa 2a. As single agent therapy in malignant melanoma, interferon alfa-2a was only marginally useful in most patients. Nevertheless, combination regimens of this agent with cytotoxic agents, alternative molecular species of interferon, and lymphokines, notably tumor necrosis factor, offer a conceptually intriguing dimension in the design of future clinical trials. PMID- 10822465 TI - Evolving therapy of hairy cell leukemia. AB - The rationale for antileukemic therapy in hairy cell leukemia is to reduce the significant risk of infection and other potential serious complications. Corticosteroids have limited value; both corticosteroids and chemotherapy are associated with substantial risks of infection. The mainstay of therapy has been splenectomy. Improvement is seen in 50% to 70% of patients with cytopenias; although the impact of splenectomy on survival has not been clearly demonstrated, prolonged hematologic improvement can occur. Splenectomy presumably alleviates the pancytopenic effect of hypersplenism by removing the preferred site of leukemic cell proliferation. Human interferon represents a major advance in management. Favorable results with natural leukocyte alpha interferon have been confirmed by data with biosynthetic (recombinant) alpha interferon. Importantly, the incidence of infection has been clearly shown to decrease, suggesting improved survival in patients with advanced hairy cell leukemia. Many questions regarding interferon therapy remain unanswered, including optimal dose, optimal duration, and maintenance therapy after maximal response. The mechanism of action is unclear, but possibly interferon modulates as yet unidentified lymphokines or growth factors. In vitro evidence suggests a direct antiproliferative effect of type I interferon on hairy cells. Preliminary data suggest that although toxicity issues, including induction of immunodeficiency and renal insufficiency require further clarification, deoxycoformycin, an adenosine deaminase inhibitor, is also highly effective and holds substantial promise as an important therapeutic modality. PMID- 10822464 TI - Interferon treatment of renal cell carcinoma. Current status and future prospects. AB - Studies with various interferon alpha preparations, including interferons induced in human leukocytes, interferon alfa-N1, interferon alfa-2a, and interferon alfa 2b, have all provided evidence for modest but reproducible antitumor activity in advanced renal cell carcinoma. Review of the data suggests that maximal response rates are achieved when interferon alpha is administered within a fairly restricted range of moderate to high doses, whereas extremely low or extremely high dosage regimens appear less likely to induce therapeutic response. Preliminary evidence suggests that interferons beta and gamma may also induce regression of metastatic renal cell carcinoma. Recent in vitro and animal studies have shown that combinations of interferon gamma with interferon alpha or interferon beta, produce synergistic biologic activities, suggesting that the various interferons may have different pathways of action related to agent specific cellular receptors. Possible interactions of different interferon species given concurrently to patients with renal cell carcinoma are under investigation, as are combinations of interferon alpha with chemotherapeutic agents. Despite in vitro data suggesting enhanced antiproliferative activity for the combination of interferon alpha and vinblastine, most clinical studies of this combination have proved to be no more effective than interferon alpha alone, but they have provided evidence of increased toxicity. The recent identification and purification of other biologically active cytokines, such as tumor necrosis factor and interleukin-2, and of monoclonal antibodies that recognize unique cell surface antigens on renal carcinoma cells, provide exciting possibilities for combination regimens with various interferon species. PMID- 10822466 TI - Current and future uses of recombinant interferon alpha in the treatment of low grade non-Hodgkin's lymphoma. AB - Recombinant and natural forms of interferon alpha have been shown to bring about tumor regressions in patients with low-grade non-Hodgkin's lymphoma. Complete and partial response rates in up to 50% of patients have been documented. Although these studies are encouraging, interferon alpha's role in the current management of patients with low-grade non-Hodgkin's lymphoma is not apparent. The high doses of interferon that were used in some of the recent studies were not well tolerated by these patients. Current studies using interferon in patients with low-grade non-Hodgkin's lymphomas are evaluating lower, and perhaps better tolerated doses of interferon. In addition, in vitro studies have documented additive and sometimes synergistic antitumor effects of interferon in combination with cytotoxic drugs. This approach is being extended to the clinic, and trials are now underway evaluating combinations of interferon and various cytotoxic drugs known from previous studies to be active in patients with low-grade non Hodgkin's lymphomas. This report reviews studies using interferon alone in these patients, and discusses ongoing and proposed studies for the future use of interferon in treating this disease. PMID- 10822467 TI - Therapy of chronic myelogenous leukemia. AB - While the demonstrated antiviral, antiproliferative, and immunomodulatory properties of interferons have led to a number of theories regarding their potential use in treating individuals with chronic myelogenous leukemia (CML), their limited availability has prevented thorough clinical investigation. However, in 1980, successful cloning of the mature human alpha-A interferon led to the production of large quantities of bacterially synthesized human alpha-A interferon, now designated interferon alfa-2a (Roferon-A, Hoffmann-La Roche, Nutley, NJ). This abundance of alfa-2a made possible clinical studies of alpha interferon's capacity to suppress CML Philadelphia (Ph1) clones as well as restore the cells with normal karyotype. The data resulting from these clinical trials indicate that interferon alfa-2a is effective in inducing hematologic remissions in the majority of minimally treated, benign-phase CML, Ph1-positive patients. In some of the patients, treatment resulted in Ph1 chromosome suppression in the bone marrow and in the emergence of cells with a normal karyotype. PMID- 10822468 TI - Incidence and clinical significance of neutralizing antibodies in patients receiving recombinant interferon alfa-2a by intramuscular injection. AB - More than 1600 patients with neoplastic disorders have received recombinant human interferon alfa-2a (Roferon-A, Hoffmann-La Roche, Nutley, NJ) as part of ongoing or completed clinical trials. In this report, the efficacy of interferon alfa-2a therapy was compared with the incidence of antibodies to this interferon in 617 patients who received the drug by intramuscular administration. Antibody measurements were performed using a highly sensitive enzyme immunoassay, and an interferon antiviral neutralization bioassay. Partial or complete remission occurred in 28% (43 of 152) of the antibody-positive patients, and in 24% (112 of 465) of the antibody-negative patients (P = 0.33). The highest incidence of antibody formation occurred among patients with renal cell carcinoma and acquired immune deficiency syndrome (AIDS)-related Kaposi's sarcoma (44% and 34%, respectively). Both the duration of treatment and length of survival were significantly longer for antibody-positive than for antibody-negative patients. No significant intergroup differences emerged for response rates or for time to onset or duration of therapeutic response. When results from the above assays were compared to those used for the detection of antibodies to recombinant interferon alfa-2b (Intron A, Schering-Plough Inc., Kenilworth, NJ), the immunoradiometric assay method was determined to be seriously deficient for determination of antibody incidence. This decreased assay sensitivity may account for the reportedly lower incidence of antibodies to recombinant alfa-2b interferon. PMID- 10822469 TI - Private health care in Canada. PMID- 10822471 TI - One fatal night. PMID- 10822470 TI - Earning a driver's license. AB - Teenage drivers in the United States have greatly elevated crash rates, primarily a result of qualities associated with immaturity and lack of driving experience. State licensing systems vary substantially, but most have allowed quick and easy access to driving with full privileges at a young age, contributing to the crash problem. Formal driver education has not been an effective crash prevention measure. Following the introduction of graduated licensing in New Zealand, Australia, and Canada, this system has been considered in many states and has been implemented in some. Graduated systems phase in full privilege driving, requiring initial experience to be gained under conditions of lower risk. The author describes the first five multistage graduated systems enacted in the United States in 1996 and 1997. Factors that will influence the acceptability and effectiveness of these new licensing systems are discussed. PMID- 10822473 TI - Position on the agreement between the State Attorneys General and the tobacco industry. American College of Preventive Medicine. PMID- 10822472 TI - Child labor still with us after all these years. AB - Child labor is a major threat to the health of children in the United States. The U.S. Department of Labor estimates that more than four million children are legally employed and that another one to two million are employed under illegal, often exploitative conditions. Across the United States, child labor accounts for 20,000 workers compensation claims, 200,000 injuries, thousands of cases of permanent disability, and more than 70 deaths each year. Agriculture and newspaper delivery are the two most hazardous areas of employment for children and adolescents. Poverty, massive immigration, and relaxation in enforcement of Federal child labor law are the three factors principally responsible for the last two decades' resurgence of child labor in the United States. Control of the hazards of child labor will require a combination of strategies including vigorous enforcement, education, and public health surveillance. PMID- 10822474 TI - "Racial" and ethnic classification: two steps forward and one step back? PMID- 10822475 TI - Can we monitor socioeconomic inequalities in health? A survey of U.S. health departments' data collection and reporting practices. AB - OBJECTIVE: To evaluate the potential for and obstacles to routine monitoring of socioeconomic inequalities in health using U.S. vital statistics and disease registry data, the authors surveyed current data collection and reporting practices for specific socioeconomic variables. METHODS: In 1996 the authors mailed a self-administered survey to all of the 55 health department vital statistics offices reporting data to the National Center for Health Statistics (NCHS) to determine what kinds of socioeconomic data they collected on birth and death certificates and in cancer, AIDS, and tuberculosis (TB) registries and what kinds of socioeconomic data were routinely reported in health department publications. RESULTS: Health departments routinely obtained data on occupation on death certificates and in most cancer registries. They collected data on educational level for both birth and death certificates. None of the databases collected information on income, and few obtained data on employment status, health insurance carrier, or receipt of public assistance. When socioeconomic data were collected, they were usually not included in published reports (except for mothers educational level in birth certificate data). Obstacles cited to collecting and reporting socioeconomic data included lack of resources and concerns about the confidentiality and accuracy of data. All databases, however, included residential addresses, suggesting records could be geocoded and linked to Census-based socioeconomic data. CONCLUSIONS: U.S. state and Federal vital statistics and disease registries should routinely collect and publish socioeconomic data to improve efforts to monitor trends in and reduce social inequalities in health. PMID- 10822476 TI - Missed opportunities in monitoring socioeconomic status. PMID- 10822477 TI - Alzheimer's disease: the evolution of a diagnosis. PMID- 10822478 TI - Alzheimer's disease as a cause of death in the United States. AB - OBJECTIVE: To describe the scope of mortality from and trends in Alzheimer's disease, to show how Alzheimer's disease ranks as a leading cause of death, to describe a methodological change regarding ranking, and to discuss issues related to the reporting of Alzheimer's disease on death certificates. METHODS: The authors analyzed mortality data from the National Vital Statistics System. RESULTS: Alzheimer's disease has increasingly been reported as a cause of death on death certificates in the United States; however, this increase may represent a variety of factors including improved diagnosis and awareness of the disease or changes in the perception of Alzheimer's disease as a cause of death. In 1995, Alzheimer's disease was identified as the underlying cause of 20,606 deaths. Overall, Alzheimer's disease was the 14th leading cause of death in 1995; for people 65 years of age or older, it was the 8th leading cause of death. Both death rates and cause-of-death ranking differed by selected demographic variables. CONCLUSIONS: In recognition of the importance of the condition as a major public health problem, Alzheimer's disease was added to the list of causes eligible to be ranked as leading causes of death in the United States beginning with mortality data for 1994. Several issues need to be kept in mind in interpreting mortality data on Alzheimer's disease, including how diagnoses are made, how the condition is classified, and the purpose of death certificates. PMID- 10822479 TI - Childhood asthma surveillance using computerized billing records: a pilot study. AB - OBJECTIVE: This paper describes a pilot project to develop and implement a low cost system for ongoing surveillance of childhood asthma in Milwaukee County, Wisconsin. METHODS: The authors organized a planning workshop to solicit information and ideas for an asthma surveillance system, bringing together national experts with Milwaukee professionals and community representatives involved in the prevention and treatment of asthma. Based on recommendations from the workshop, a pilot surveillance project was implemented in Milwaukee County using records of emergency room visits and hospital admissions for asthma abstracted from the computerized billing records of the Children's Hospital of Wisconsin (CHW), retrospectively for 1993 and prospectively for 1994. Retrospective data were also sought from the other hospital emergency departments in Milwaukee County to evaluate the representativeness of the CHW data. Surveillance data were used to evaluate utilization of care by patient subgroups and to describe temporal patterns in emergency room visits. RESULTS: Of the emergency department visits for asthma in Milwaukee County in 1993, CHW accounted for 94% among infants less than 1 year of age, 89% among children ages 1 through 5 years, and only 59% among children between the ages of 6 and 18 years. In 1994, the 7% of asthmatic children with repeat hospital admissions accounted for 38% of all hospital admissions for asthma and the 20% with repeat emergency department visits accounted for 50% of all emergency visits. Emergency visits for asthma showed clear seasonality, with a peak in the fall and a smaller peak in the spring. CONCLUSIONS: Computerized medical billing data provide an opportunity for asthma surveillance at a relatively low cost. The data obtained are useful for tracking trends in exacerbations of asthma and the use of medical services for asthma care and should prove valuable in targeting interventions. PMID- 10822481 TI - Before the Surgeon General: marine hospitals in mid-19th-century America. PMID- 10822480 TI - Effectiveness and cost of rapid and conventional laboratory methods for Mycobacterium tuberculosis screening. AB - OBJECTIVE: Because delay in the diagnosis of tuberculosis (TB) contributes to the spread of disease and the associated mortality risk, the authors examined the effectiveness and cost of recent advances in methods of diagnosing TB and testing for drug susceptibility, comparing these rapid methods to traditional approaches. METHODS: Decision analysis was used to compare newer rapid and older nonrapid methods for diagnosing TB and testing for drug susceptibility. The average time to diagnosis, average time to treatment, average mortality, and cost of caring for patients evaluated for TB were compared. RESULTS: Using a combination of solid medium and broth cultures, nucleic acid probes for identification, and radiometric broth drug susceptibility testing would lead to diagnosis on average 15 days faster and to appropriate therapy on average five days sooner than methods currently employed by many U.S. laboratories. The average mortality would drop by five patients per 1000 patients evaluated (31%) and the average cost per patient would drop by $272 (18%). CONCLUSIONS: In this era of cost containment, it is important to incorporate test sensitivity and specificity when evaluating technologies. Tests with higher unit costs may lead to lower medical expenditures when diagnostic accuracy and speed are improved. U.S. laboratories should employ available rapid techniques for the diagnosis of TB. PMID- 10822482 TI - Improved osteoconduction of cortical bone grafts by biodegradable foam coating. AB - Alteration of the geometrical surface configuration of cortical bone allografts may improve incorporation into host bone. A porous biodegradable coating that would maintain immediate structural recovery and subsequently allow normal graft healing and remodeling by promoting bony ingrowth could provide an osteoconductive surface scaffold. We investigated the feasibility of augmenting cortical bone grafts with osteoconductive biodegradable polymeric scaffold coatings. Three types of bone grafts were prepared: Type I--cortical bone without coating (control), Type II--cortical bone coated with PLGA-foam, Type III- cortical bone coated with PPF-foam. The grafts were implanted into the rat tibial metaphysis (16 animals for each type of bone graft). Post-operatively the animals were sacrificed at 2 weeks and 4 weeks (8 animals for each type of bone graft at each time point). Histologic and histomorphometric analysis of grafts showed that the amount of new bone forming around the foam-coated grafts was significantly higher than in the control group (uncoated; p < 0.02). Although both foam formulations were initially equally osteoconductive, PLGA-based foam coatings appeared to have degraded at two weeks postoperatively, whereas PPF-based foam coatings were still present at 4 weeks postoperatively. While significant resorption was present in control allografts with little accompanying reactive new bone formation, PLGA-coated bone grafts showed evidence of bone resorption and subsequent bony ingrowth earlier than those coated with PPF-based foams suggesting that PPF-coated cortical bone grafts were longer protected against host reactions resulting in bone resorption. PMID- 10822483 TI - Effect of alpha/gamma phase ratio on corrosion behavior of dual-phase stainless steels. AB - Dual-phase stainless steels have been developed in order to reduce the nickel content, which is potentially responsible to an allergic reaction when these steels are used as medical or dental applications. In this study, two different dual-phase stainless steels (2205 and Z100) were electrochemically tested to evaluate their corrosion resistance in three corrosive solutions (i.e., synthetic saliva, 0.9% NaCl solution, and Ringer solution). Particularly, an attempt was made to correlate the corrosion resistance to a metallographic parameter, which is, in this study, the alpha/gamma phase ratio. It was concluded that (1) type 2205 stainless steel exhibited excellent corrosion resistance in all three corrosion media; however 2205 stainless steel decreases its corrosion resistance by increasing chloride concentration in tested electrolytes from synthetic saliva through 0.9% NaCl solution to Ringer solution. (2) X-ray diffraction analysis indicated that the alpha/gamma phase ratio of 2205 (1.735) was higher than that of Z100 (0.905). As a result, it is suggested that by increasing the alpha/gamma phase ratio the material shows more corrosion-prone behavior when being subjected to a hostile environment containing higher chloride ion concentration. PMID- 10822484 TI - Kinematic and dynamic analysis of running under conditions of variable gravity. AB - A new training system was designed and realised by the engineer G. Scuderi: a centrifugal track for runners. The principal advantage of this track is to increase the forces on athlete during the run with an effect very similar to that of an "artificial gravity", so the athlete can develop more muscle power. A multibody numerical model of a runner was developed to analyse the behaviour of the athlete on the centrifugal track. The multibody model was calibrated by experimental analysis; in fact, the joint angles were measured by a digital image processing system and introduced as input in the numerical simulation. The numerical results obtained are the values of many kinematic and dynamic variables, in particular the ground reaction force and the moments in the hip, knee, and ankle joints. A comparison between the normal running and that on the centrifugal track was carried out, demonstrating the validity of training on the latter. The results of the numerical simulation confirm that the power developed during running on the track, and the corresponding work performed by the joints, is considerably greater than that found during normal running. PMID- 10822485 TI - Sandwiched osmotic tablet core for nifedipine controlled delivery. AB - The sandwiched osmotic tablet core, which is composed of a middle push layer and two attached drug layers, has been prepared and systematically studied with the purpose of delivering water-insoluble nifedipine. The advantage of the sandwiched osmotic tablet system over the commercialized push-pull osmotic tablet system is its simplicity of preparation, as the surface identification was avoided. It was observed that polyethylene oxides (PEO) with molecular weight (MW) of 300,000 and 8,000,000 g/mole were suitable for the thickening agent of drug layer and the expandable hydrogel of push layer, respectively. The weight ratio of 190/190 for drug layer/push layer was also found to be suitable. It has been observed that PEO amount of the push layer and the KCl amount of the drug layer had profoundly positive influence on nifedipine release. A push layer/drug layer co-controlled osmotic delivery mechanism has been proposed and the optimal tablet formulation has been obtained. It was also found that PEO and nifedipine were miscible, which may support the application of PEO in nifedipine dosage forms. Meanwhile, the PEO/nifedipine binary phase diagram has been constructed. The sandwiched osmotic tablet system can deliver nifedipine in an approximate zero-order rate up to 24 hours. It may be potentially used for the delivery of water-insoluble drugs. PMID- 10822487 TI - Biomechanical behavior of hydroxyapatite as bone substitute material in a loaded implant model. On the surface strain measurement and the maximum compression strength determination of material crash. AB - Many investigators have advocated that hydroxyapatite ceramics may be extremely prospective bone substitute material mainly through evidence with its given biocompatibility towards bone and demonstrated continuity between living bone and hydroxyapatite. Its mechanical brittleness and strength, however, have been the most serious considerations. In this paper, a study has been performed using animal experiments, massive hydroxyapatite ceramics are implanted into rectangular bone defect created by operation at the load-bearing area in close contact with tibia plateau. The changes of surface strain at the hydroxyapatite implantation of retrieved tibiae are measured at any week after implantation when the compressive stress were applied in the direction of long axis in order to clarify the mechanical behaviors of hydroxyapatite living bone complex. The compression strength of hydroxyapatite implant crash was also determined at each week after implantation. Results revealed that the hydroxyapatite living bone complex has been proven to have sufficient flexibility such that it shows no hysteresis in stress versus strain relationship up to 200 kg of applied load. The strain pattern on the surface of hydroxyapatite implant develops quite similar to that of natural cortical bone. The material crash of hydroxyapatite implant tolerates up to 500 kg of load at 52 weeks after implantation. These characteristics suggest that it achieves normal skeletal function in the points of biomechanical properties in vivo. PMID- 10822486 TI - Residual strain in human atherosclerotic coronary arteries and age related geometrical changes. AB - The opening angles of 16 rings excised from human coronary arteries with different degrees of atherosclerosis were determined 10 hours after death. Atherosclerosis, as a chronic inflammatory response of arterial endothelium and intima, is defined by three degrees of its development. The opening angle decreases more or less linearly with the distance from the right coronary artery orifice. This is in accordance with the functional requirements posed on blood transport into the coronary arteries. A decrease of the opening angle with age is affected by hardening of the arterial wall, among other factors. This is in accordance with a stochastic model of age related changes in the initial modulus of elasticity of the coronary artery. A part of the free energy of smooth muscle cells, fibroblasts, collagen and elastic fibres is used not for creating residual strain but for remodeling the arterial wall structure. The opening angle is also considerably affected by the degree of atheroclerosis. The dependence on age of the external diameter and the thickness of the intact left and right coronary arteries in the vicinity of the aortic sinus was also analyzed in two female and two male subjects. To ensure the objectivity of the results it is necessary to carry out additional experiments and studies in vivo. PMID- 10822488 TI - Augmentation of osteoinduction with a biodegradable poly(propylene glycol-co fumaric acid) bone graft extender. A histologic and histomorphometric study in rats. AB - We investigated the feasibility of enhancing the regeneration of skeletal tissues by augmenting bone grafts with a composite biodegradable bone graft extender material based on the polymer poly(propylene fumarate), PPF. The material was mixed with autograft and allograft and placed directly into a cylindrical metaphyseal defect made in the rat tibia. These formulations were compared to defects without any graft material, autografts, allografts and PPF alone. Nine animals were included in each group. Animals were sacrificed at 1 and 4 weeks postoperatively. Implantation sites were then evaluated using histologic and histomorphometric methods. Results of this study showed that defects did not heal in sham operated animals. In the experimental groups, there was early new woven bone formation in the autograft group with near complete healing of the defect at four weeks. When PPF was used alone, gradual ingrowth of new bone was seen. Mixing of the PPF bone graft extender with either allograft or autograft material resulted in enhancement of new bone formation with both allo- and autograft. However, significantly more new bone formation than in the autograft group was only seen when the PPF bone graft extender was mixed with fresh autograft. Histomorphometry corroborated these findings. Results of this study suggest that a PPF-based material may be used to increase the volume of smaller amounts of bone grafts supporting the concept of "bone graft extenders" by application of engineered biodegradable porous scaffolds. PMID- 10822489 TI - Modeling of occupant dynamics during automobile rear-end impacts. AB - Procedures for studying the dynamic response of the occupant within a rear-end impacted vehicle are presented. Most of the researches in the impact analysis were performed by experimental approach and this costs a lot of time and money. Especially, the repeatability is very hard to produce in a destructive condition. Most of all, the analytic parameters can be investigated are limited by the experimental approach. By using numerical techniques, this research employs Kane's equation and Huston's method to develop a simulated system with visual graphic output to observe the rear-end impact response. According to the simulated results, at a constant seatback angle the maximum acceleration values of head and chest increased with the increasing of impact velocity. Furthermore, at a constant impact velocity the relative rotation angle of a passenger's head to chest decreased with the increasing of initial seatback angle. PMID- 10822490 TI - The mechanical-chemical attachment between the artificial articular cartilage (PVA-hydrogel) and metal substrate (or underlying bone). AB - PVA-hydrogel has excellent biocompatibility as well as preferable tribological characteristics as an artificial articular cartilage. One of the most difficult problems is the attachment of PVA-hydrogel to the underlying bone (or metal substrate). In the present study, the micro-mechanical attachment between the PVA hydrogel and metal fibre mesh was at first accomplished to obtain a composite artificial cartilage device (CACD), then the surface of fibre mesh is chemically bonded to the metal substrate (or underlying bone) by adhesive (PMMA). This method can be used to accomplish mechanical-chemical attachment between composite artificial cartilage device and underlying bone (or metal substrate). Microstructure analysis and mechanical tests indicate that the CACD can be firmly bonded to the metal substrate (or underlying bone). PMID- 10822491 TI - Nuclear plasmids of Dictyostelium. PMID- 10822492 TI - The translation initiation signal in E. coli and its control. PMID- 10822493 TI - Direct isolation of specific chromosomal regions and entire genes by TAR cloning. PMID- 10822494 TI - Genetic engineering of plant chilling tolerance. PMID- 10822495 TI - Role of bacterial chaperones in DNA replication. AB - Studies on the involvement of chaperone proteins in DNA replication have been limited to a few replication systems, belonging primarily to the prokaryotic world. The insights gained from these studies have substantially contributed to our understanding of the eukaryotic DNA replication process as well. The finding that molecular chaperones can activate some initiation proteins before DNA synthesis has led to the more general suggestion that molecular chaperones can influence the DNA-binding activity of many proteins, including transcriptional factors involved in cell regulatory systems. The DnaK/DnaJ/GrpE molecular chaperone system became a paradigm of our understanding of fundamental processes, such as protein folding, translocation, selective proteolysis and autoregulation of the heat-shock response. Studies on the Clp ATPase family of molecular chaperones will help to define the nature of signals involved in chaperone dependent proteins' refolding and the degradation of misfolded proteins. PMID- 10822496 TI - Mechanisms of initiation of linear DNA replication in prokaryotes. PMID- 10822497 TI - Diverse regulatory mechanisms of amino acid biosynthesis in plants. PMID- 10822498 TI - Proton transport by bacteriorhodopsin in planar membranes assembled from air water interface films. AB - Bacteriorhodopsin, in known amounts and controlled orientation, is incorporated into planar membrane films. These films are formed by the sequential transfer of two air-water interface films onto a thin, hydrophilic, electrically conductive support cast from nitrocellulose. The films are easily accessible to electrical measurements and to control of the ionic milieu on either side of the membrane. The area of the assembled membrane films can be varied between 2.3 x 10(-2) cm2 and 0.7 cm2. Illumination of these films produces photocurrents, photovoltages, and changes in the pH of the surrounding medium. The peak amplitude of the photocurrent increases linearly with light intensity for dim lights, and it approaches a saturating value for brighter lights. In the linear range, the stoichiometry of transport is 0.65 +/- 0.06 protons/absorbed photon. The rate of transport is linearly proportional to light at all intensities tested. The amplitude and kinetics of the photovoltage measured are accurately predicted by the photocurrent generated and the passive electrical features of the film. Parallel measurements of pH and photocurrent reveal that the light-induced changes in pH are fully accounted for by the rate and amount of charge transport across the membrane. Preceding the transport of protons, a transient photovoltage is detected that exhibits no detectable latency, reaches peak in about 80 microseconds, and probably arises from light-induced intramolecular charge displacements. PMID- 10822499 TI - Volume regulation by Amphiuma red blood cells. The membrane potential and its implications regarding the nature of the ion-flux pathways. AB - After osmotic perturbation, the red blood cells of Amphiuma exhibited a volume regulatory response that returned cell volume back to or toward control values. After osmotic swelling, cell-volume regulation (regulatory volume decrease; RVD) resulted from net cellular loss of K, Cl, and osmotically obliged H2O. In contrast, the volume-regulatory response to osmotic shrinkage (regulatory volume increase; RVI) was characterized by net cellular uptake of Na, Cl, and H2O. The net K and Na fluxes characteristic of RVD and RVI are increased by 1-2 orders of magnitude above those observed in studies of volume-static control cells. The cell membrane potential of volume-regulating and volume-static cells was measured by impalement with glass microelectrodes. The information gained from the electrical and ion-flux studies led to the conclusion that the ion fluxes responsible for cell-volume regulation proceed via electrically silent pathways. Furthermore, it was observed that Na fluxes during RVI were profoundly sensitive to medium [HCO3] and that during RVI the medium becomes more acid, whereas alkaline shifts in the suspension medium accompany RVD. The experimental observations are explained by a model featuring obligatorily coupled alkali metal H and Cl-HCO3 exchangers. The anion- and cation-exchange pathways are separate and distinct yet functionally coupled via the net flux of H. As a result of the operation of such pathways, net alkali metal, Cl, and H2O fluxes proceed in the same direction, whereas H and HCO3 fluxes are cyclic. Data also are presented that suggest that the ion-flux pathways responsible for cell-volume regulation are not activated by changes in cell volume per se but by some event associated with osmotic perturbation, such as changes in intracellular pH. PMID- 10822500 TI - Some properties of potassium-stimulated calcium influx in presynaptic nerve endings. AB - Potassium-stimulated 45Ca entry into rat brain synaptosomes was measured at times ranging from 1 to 60 s. The K-rich solutions were used to depolarize the synaptosomes. Backflux of 45Ca from the synaptosomes was negligible during the first 10-20 s of incubation. An initial ("fast") phase of K-stimulated Ca entry, lasting from 1 to 2 s was observed. This phase was inhibited by low concentrations of La (KI approximately equal to 0.3 microM). It was also abolished ("inactivated") by incubating the synaptosomes in depolarizing solutions (containing veratridine, gramicidin, or elevated [K]o) before the addition of 45Ca. An additional long lasting ("slow") phase of K-stimulated Ca entry was also detected. This "slow" Ca entry was much less sensitive to La (KI > 100 microM) and was not affected by depolarizing the synaptosomes before the addition of 45Ca. The rate of influx during the fast phase was about four times the rate of Ca influx during the slow phase. Neither the fast nor slow phase of Ca entry was sensitive to tetrodotoxin (10 microM), a potent blocker of Na channels, but both phases were inhibited by Ni, Mn, Mg, and other agents that block Ca channels. The data are consistent with the presence of two distinct populations of voltage-regulated, divalent cation-selective pathways for Ca entry in presynaptic brain nerve endings. PMID- 10822501 TI - Action potentials of isolated single muscle fibers recorded by potential sensitive dyes. AB - Light transmission changes upon massive stimulation of single muscle fibers of Xenopus were studied with the potential-sensitive nonpermeant dyes, merocyanine rhodanine (WW375) and merocyanine oxazolone (NK2367). Upon stimulation an absorption change (wave a) occurred, which probably represents the sum of action potentials in the transverse tubules and surface membrane. In WW375-stained fibers wave a is a decrease in transmission over the range of 630 to 730 nm (with NK2367, over the range of 590 to 700 nm) but becomes an increase outside this range, thus showing a triphasic spectral pattern. This spectrum differs from that of the squid axon, in which depolarization produces only an increase in transmission over the whole range of wavelengths (Ross et al. 1977. J. Membr. Biol. 33:141-183). When wave a was measured at the edge of the fiber to obtain more signal from the surface membrane, the spectrum did not seem to differ markedly from that obtained from the entire width of the fiber. Thus, the difference in the spectrum between the squid axon and the vertebrate muscle cannot be attributed to the presence of the tubular system. PMID- 10822502 TI - Radial propagation of muscle action potential along the tubular system examined by potential-sensitive dyes. AB - Isolated single (Xenopus) muscle fibers were stained with a non-permeant potential-probing dye, merocyanine rhodanine (WW375) or merocyanine oxazolone (NK2367). When the fiber was massively stimulated, an absorption change (wave a), which seemed to reflect the action potential, occurred. Simultaneous recording of optical changes and intracellular action potentials revealed that the time-course of wave a was slower than the action potential: the peak of wave a was attained at 1 ms, and the peak of action potential was reached at 0.5 ms after the stimulation. This difference suggests that wave a represents the potential changes of the whole tubular membrane and the surface membrane, whereas the action potential represents a surface potential change. This idea was substantiated by recording absorption signals preferentially from the surface membrane by recording the absorption changes at the edge of the fiber. Wave a obtained by this method was as quick as the intracellular action potential. The value of radial conduction velocity of action potential along the T system, calculated by comparing the action potential with wave a, was 6.4 cm/s at 24.5 degrees C, in fair agreement with Gonzalez-Serratos (1971. J. Physiol. [Lond.]. 212:777-799). The shape of wave a suggests the existence of an access delay (a conduction delay at the orifice of the T system) of 130 microseconds. PMID- 10822503 TI - Cone contributions to cat retinal ganglion cell receptive fields. AB - Extracellular microelectrode recordings were made from ganglion cells of the intact, in situ eyes of adult common domestic cats. Three different photopic systems, with peak spectral sensitivities at 450, 500, and 556 nm, were observed. All ganglion cells received input from a cone system with a peak spectral sensitivity of 556 nm. The blue-sensitive cone system was observed in about one half of the ganglion cells studied. In each case the 450-nm cone system contributed to only one functional type of response, either ON or OFF, in the same cell. The other two photopic systems most often contributed to both the ON and OFF responses of an individual ganglion cell. In four cases the 450-nm cone system mediated responses that were opponent to those of the other two photopic systems. The third photopic mechanism has a peak spectral sensitivity at 500 nm and contributed to most receptive field surrounds and many receptive field centers. It is distinguished from the rod system by the occurrence of a break in both dark-adaptation curves and increment-sensitivity curves. No apparent differences in receptive field cone contributions between brisk-sustained and brisk-transient cells were seen. PMID- 10822505 TI - Culture filtrates of Aspergillus fumigatus induce different modes of cell death in human cancer cell lines. AB - Aspergillus fumigatus culture filtrate (CF) has a potent cytotoxic effect on three human cancer cell lines (DLKP, A549 and HEp-2) and initiates cell death by apoptosis but the execution of the apoptotic process is incomplete. DLKP cells treated with A. fumigatus CF demonstrate features associated with apoptosis but cytoplasmic and nuclear fragmentation were not observed and cells ultimately underwent necrosis. The apoptotic process commenced in A549 and HEp-2 cells upon exposure to CF, cell shrinkage was observed but membrane blebbing and apoptotic body formation were not detected and detached cells died by necrosis. In contrast, extensive nuclear fragmentation and apoptotic body formation were evident in DLKP and A549 cells treated with anti-neoplastic agents. This work indicates that A. fumigatus CF is cytotoxic to cancer cells and can initiate apoptosis but that the complete apoptotic pathway is not followed. PMID- 10822506 TI - Mitochondrial DNA analysis of Exophiala jeanselmei var. lecanii-corni and Exophiala castellanii. AB - A Japanese clinical isolate (KU-A-0094) which was identified by de Hoog et al. as Exophiala jeanselmei var. lecanii-corni with difficulty, was compared with 5 strains including the type cultures of E. jeanselmei var. lecanii-corni, var. jeanselmei and E. castellanii using RFLP (restriction fragment length polymorphism) patterns of mtDNA (mitochondrial DNA). RFLP patterns of KUA-0094 were identical with those of E. jeanselmei var. lecanii-corni and different from those of E. castellanii with restriction enzymes of HaeIII, MspI and hindIII. Therefore, de Hoog et al.'s identification of KU-A-0094 was confirmed. Additionally, mtDNA-RFLP patterns of E. jeanselmei var. lecanii-corni and E. jeanselmei var. jeanselmei were also different from each other. Consequently E. jeanselmei var. lecanii-corni seem to be a species in its own right rather than a variant of E. jeanselmei. PMID- 10822504 TI - Murine models of blastomycosis, coccidioidomycosis, and histoplasmosis. AB - Animal models have contributed much to the knowledge of fungal infections and their corresponding therapeutic treatments. This is true for animal models of the primary fungal pathogens, Blastomyces dermatitidis, Coccidioides immitis, and Histoplasma capsulatum. This review gives a brief background of human diseases associated with these organisms and describes the development, details, and utility of murine models of blastomycosis, as well as coccidioidomycosis and histoplasmosis. PMID- 10822507 TI - Evaluation of the susceptibility of Prototheca zopfii to milk pasteurization. AB - Protothecosis has been reported in humans (gastroenteritis, bursitis, etc.) and in many other animal species. Bovine mastitis represents the main form of occurrence of protothecosis in cattle. Milk as well as dairy products, when contaminated with Prototheca spp., represent a potential means of transmission of this zoonosis. The purpose of this study was to evaluate the susceptibility of forty Prototheca zopfii strains isolated from milk from intramammary infections in dairy cows and also from bulk milk tanks of dairy farms, to the different ratios of temperature/time employed in the thermal treatment of milk: 72-75 degrees C/15 seconds, 72-75 degrees C/20 seconds and 62-65 degrees C/30 minutes. The samples were subjected to these different temperature/time ratios. The evaluation of the thermal susceptibility of the P. zopfii strains showed that 34 strains were resistant in at least one of the tests. The results point out the need to consider the importance of mastitis caused by Prototheca spp. as representing a public health risk. PMID- 10822508 TI - Screening of fungal species for fumonisin production and fumonisin-like disruption of sphingolipid biosynthesis. AB - Fumonisins are mycotoxins produced by several species of Fusaria. They are found on corn and in corn-based products, can cause fatal illnesses in some animals and are suspected human esophageal carcinogens. Fumonisins are believed to cause toxicity by blocking ceramide synthase, a key enzyme in sphingolipid biochemistry which converts sphinganine (or sphingosine) and fatty acyl CoA to ceramide. Relatively few fungal species have been evaluated for their ability to produce fumonisins. Fewer have been studied to determine if they produce ceramide synthase inhibitors, whether fumonisin-like structures or not, therefore potentially having toxicity similar to fumonisins. We analyzed corn cultures of 49 isolates representing 32 diverse species of fungi for their ability to produce fumonisins. We also evaluated the culture extracts for ceramide synthase activity. Only cultures prepared with species reported previously to produce fumonisins--Fusarium moniliforme and F. proliferatum--tested positive for fumonisins. Extracts of these cultures inhibited ceramide synthase, as expected. None of the other fungal isolates we examined produced fumonisins or other compounds capable of inhibiting ceramide synthase. Although the fungi we selected for these studies represent only a few of the thousands of species that exist, they share the commonality that they are frequently associated with cereal grasses, including corn, either as pathogens or as asymptomatic endophytes. Thus, these results should be encouraging to those attempting to find ways to genetically manipulate fumonisin-producing fungi, to make corn more resistant, or to develop biocontrol measures because it appears that only a relatively few fungal contaminants of corn can produce fumonisins. PMID- 10822509 TI - The occurrence of ochratoxin A in dust collected from a problem household. AB - Accumulated dust samples were collected from the heating ducts in a household where signs resembling ochratoxin poisoning in animals occurred. Several Penicillium spp. and Aspergillus ochraceous had been identified previously from air samples taken from this house. A composite sample from six collected samples was examined by HPLC, and it was determined that 58 ppb of ochratoxin A was present in this sample. A second set of six samples was collected and determinations were made by HPLC of the ochratoxin content in each sample. All samples, including one sample of dirt from a crawl space, yielded at least a trace of ochratoxin A; however, one sample of dust collected from the heating ducts yielded over 1500 ppb of ochratoxin A, and another sample of dust from a different heating duct yielded 306 ppb of ochratoxin A. Ochratoxin A was confirmed in all samples by LC-MS, and ochratoxin was evident in the samples by TLC analysis. This is believed to be the first report of finding ochratoxin in house dust. PMID- 10822510 TI - Trimetazidine in the myocardial cell mechanisms of cytoprotection. AB - The experimental results demonstrate that TMZ displays the pharmacological characteristics of a "cytoprotective agent", since this drug exerts an anti ischemic effect on myocardium totally independent of any hemodynamic changes. PMID- 10822511 TI - Gastro-intestinal lesions induced by non-steroidal anti-inflammatory drugs. AB - Long-term use of non-steroidal anti-inflammatory drugs (NSAIDs) is associated with serious and sometimes fatal side effects as gastrointestinal ulcers, bleeding and less frequently kidney and liver damage. These side effects are caused primarily by the reduction of prostaglandin synthesis, which in turn deprives the stomach's self-protection mechanism. NSAIDs prevent prostaglandin formation by inhibiting the enzyme cyclooxygenase. Recent research has shown that there are two types of cyclooxygenase, one that produces the form of prostaglandin involved in the gastric protection and the other that produces the form of prostaglandin involved in inflammation. Current NSAIDs inhibit both forms of cyclooxygenase. Gastrointestinal lesions associated with NSAIDs are predominantly observed in the stomach and are defined under the term of "NSAID gastropathy". Intestinal lesions secondary to the ingestion of NSAIDs were only recently described. Rectocolic lesions induced by NSAID are frequent and severe. The noxious effect of NSAID on preexisting low pathologies can be differentiated from "de novo" induced acute and chronic lesions. PMID- 10822512 TI - Ischemic preconditioning during successive exercise testing. AB - It was suggested recently that ischemic preconditioning can occur in clinical practice. We investigated this hypothesis in 26 patients with old myocardial infarction (MI) or stable angina pectoris, subjected to two successive exercise testings at 1 hour interval. The ST depression was significantly lesser during the second exercise testing (1.83 +/- 0.12 mm, vs 1.02 +/- 0.14 mm p < 0.01) at the same double product (DP) (24.877 +/- 1206 vs 24.711 +/- 1152 p > 0.05) and peak effort (76.92 +/- 6.63 w vs 75.96 +/- 6.27 w p > 0.05). The improvement was attributed to ischemic preconditioning. PMID- 10822513 TI - Peripheral leukocytes activation in aortic aneurysmal disease. AB - The pathogenesis of aneurysmal disease involves factors acting over time. A sustained chronic inflammatory reaction is observed in association with initiations, maintenance, rapid growth and rupture of aortic aneurysms. This study was designed to identify the possible pathogenic role of the inflammatory cells in the outcome of aneurysmal disease, testing the activation state of peripheral lymphocytes and neutrophils. Circulating activated lymphocytes and repeated peaks of neutrophils activation in sequential follow-up is associated with larger aneurysms, mural thrombosis, tendency to aneurysm extension and rupture. PMID- 10822514 TI - Myocardial fibres in pulmonary veins--possible compensatory factor in dilated cardiomyopathy. AB - It is well known in the literature that myocardial tissue is present in different degrees in pulmonary veins, but the eventual role of this myocardial tissue in pulmonary veins is not very clear at this moment. We presumed that one of the compensatory mechanisms in heart failure could be represented by the development of this myocardial tissue in the pulmonary veins. We studied the extension of this myocardial tissue in patients with dilated cardiomyopathy (DCM). We performed a histologic examination of the myocardial tissue from the pulmonary veins, in patients with heart failure in advanced stages. These patients died in the hospital and we obtained the pulmonary tissue for examinations during the autopsy. At the border between the media and the adventitia of the pulmonary vein, the myocardial tissue is present and is well developed. Some of the fibers have a sinuous course, others present a longitudinal aspect, especially in the external layers. In some sections bundles and bands of muscle fibers are present. In the sections where no myocardial tissue was shown, the smooth muscular fibers were very numerous. The necroptic histologic studies showed different degrees of hypertrophy of the myocardial tissue in the pulmonary veins, in the patients with heart failure in advanced stages. We observed that dilatation degree of cardiac cavities and decrease of the ejection fraction correlates with hypertrophy degree of the myocardial tissue in the pulmonary veins. This can be an argument for the compensatory role of this hypertrophy. PMID- 10822515 TI - Accelerated streptokinase--a new thrombolytic regimen in acute myocardial infarction. AB - Efficiency and safety of an accelerated regimen of streptokinase (1.5 M.U. over 20 min., 109 patients) has been compared with the standard regimen (1.5 M.U. over 60 min, 119 patients) in 218 patients admitted within the first 6 hours after the onset of the symptoms of acute myocardial infarction. Using the noninvasive criteria we found a coronary reperfusion rate of 77.04% in patients belonging to the accelerated regimen group and this value was significantly higher than the one of 57.14% registered in the standard group. No major hemorrhagic events were registered in both groups. Although the hypotension appeared to be more frequent in patients in whom the accelerated regimen was used, however this side effect proved to be transient and well controlled using the rapid infusion of natrium chloride solution. In conclusion, the accelerated regimen of streptokinase is safe and followed by a higher rate of coronary reperfusion as compared to the standard one. PMID- 10822516 TI - The effect of early perindopril administration on mortality and morbidity in patients with high risk of acute myocardial infarction. AB - The angiotensin-converting enzyme inhibitors (ACEI) therapy reduces mortality and morbidity after acute myocardial infarction (AMI). Though, there are a few questions to face, in what concerns the ACEI therapy: patients selection, starting time and duration of treatment, pharmacological profile of the various ACEI and their therapeutic efficiency. We aimed to determine if early Perindopril administration in AMI, for three months, may decrease mortality and morbidity in selected patients (high risk and streptokinase thrombolysis). PMID- 10822517 TI - Results of sotalol (Darob) therapy in supraventricular tachyarrhythmias. AB - The benefit of sotalol therapy in atrial flutter and in atrial fibrillation was considered, aiming as well the suppression of paroxysmal episodes and the prevention of their recurrencies and the control of the ventricular rhythm in chronic cases. The prospective study comprises 60 patients institutionalized in the Cardiology Clinic in which the supraventricular tachyarrhythmia range included: atrial flutter (20 cases) and atrial fibrillation (40 cases), from which atrial fibrillation of recent onset (20 cases) and chronic atrial fibrillation with fast ventricular response (20 cases). Sotalol was administered orally in doses of 80 to 320 mg daily, b.i.d. The conversion to sinus rhythm was obtained in 45% of cases of atrial fibrillation of recent onset and in 30% of cases of atrial flutter. The prophylactic administration of sotalol on long term (6 months), in medium dose of 160 mg daily, may protect against the recurrence of the supraventricular arrhythmias (63.85%). As adverse effect, a case of "torsade des pointes" was recorded. IN CONCLUSION: 1) a greater benefit was noticed by using sotalol prophylactically for the recurrence of the supraventricular arrhythmias (63.85%) compared to the positive results in the therapy of acute cases (37.5%); 2) the maximal efficiency in the prevention of arrhythmia recurrencies can be observed in patients that present underlying compensated ischemic cardiopathy; 3) a decrease of 30% of the ventricular rhythm in patients with atrial fibrillation was obtained. PMID- 10822518 TI - The efficacy of sublingual verapamil in controlling rapid ventricular rate in chronic atrial fibrillation. AB - The efficacy of sublingual verapamil (Verapabene, 40 mg) in acute control of high ventricular rate was assessed in 20 patients with chronic atrial fibrillation (AF) of ischemic origin (class NYHA II and III). The effect on irregularity of the rhythm was also studied using time domain parameters of heart rate variability. Four ECG recordings (before, and at 10, 30 and 60 minutes after the administration), of 10 minutes each, were performed in every patient in basal conditions. The parameters were compared using paired t-test and Wilcoxon's test. Mean heart rate became significantly lower (p = 0.0064) after 10 minutes, and reached progressively the lowest value after 60 minutes (from the initial 112.9 beats/min to 90.6 beats/min, after 60 minutes, p < 0.0001). The irregularity parameters (SDNN--from 114.7 ms to 148.6 ms, rMSSD--from 152 ms to 205.9 ms) and the NNmin interval (from 353 ms to 404.5 ms) increased significantly after 30 and 60 minutes (p < 0.0001), but the frequency corrected irregularity (coefficient of variability) remained unchanged (p > 0.4). Sublingually administered verapamil proved to be an efficient alternative in acute control of rapid ventricular rate in AF. The method has the advantage to be applicable by a properly instructed patient itself. PMID- 10822519 TI - Treatment with alpha-interferon in chronic hepatitis. AB - The treatment of chronic viral hepatitis with Interferon has been introduced in clinical practice over the past decade an represents an important step in the management of those diseases. The data existing in literature are conflicting about the dose and period of treatment with Interferon, many treatment schedules being proposed. There are also a lot of markers used or proposed to be used to determine the response to treatment, their predicting efficacy being largely studied. The response to alpha-Interferon in a standard 6 month regimen in chronic hepatitis B, chronic hepatitis C and chronic hepatitis B and C is studied; alanine-aminotransferase (ALAT) and aspartate-aminotransferase (ASAT) are the markers used to determine the response to treatment. PMID- 10822520 TI - Hostility in patients with chronic constipation. AB - Personality changes have been reported in chronic constipation. Hostility is an important personality factor involved in psychosomatic disorders. The aim of this study was to investigate hostility in patients with chronic constipation. Sixty subjects with chronic constipation (24 males, 36 females, mean age 44.5 years) were investigated with the hostility scale of the Minnesota Multiphasic Inventory. The patients were divided in four groups according to their symptoms: functional chronic constipation (Group I, n = 18), irritable bowel syndrome expressed as chronic constipation and abdominal pain (Group II, n = 21), irritable bowel syndrome expressed as chronic constipation, abdominal pain and bloating (Group III, n = 13) and irritable bowel syndrome expressed as chronic constipation alternating with episodes of diarrhoea (Group IV, n = 8). Twenty five clinically healthy subjects were investigated as controls. Hostility was as follows (mean +/- SD): 68 +/- 9 in group I, 62 +/- 12 in group II, 70 +/- 14 in group III, 56 +/- 12 in group IV and 40 +/- 12 in controls. The scores were significantly higher in all groups of patients with constipation versus controls (p < 0.01; < 0.001; < 0.001; < 0.02, respectively). These data suggest that hostility is increased in patients with chronic constipation. It is rather a feature of the functional bowel disorders than of constipation, as symptom, only. PMID- 10822521 TI - Antioxidant protection in collagen-vascular diseases. AB - Alteration of redox balance in the serum of patients with systemic lupus erythematosus (SLE) and systemic vasculitis (SV) was investigated. Excess in oxidative processes has been measured through concentration of lipid peroxides which was found to increase by 26% in SLE and 32% in SV. Antioxidant protection capacity against this oxidative aggression has been assessed both by determining the level of activity of the enzymes participating in this process (superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase GSH-Px) and by determining the total antioxidant serum activity. The results have shown that, within antioxidant protection of the body against oxidative stress, glutathione peroxidase plays the most important role and its activity is significantly affected by the great concentration of lipidic peroxides. We have also shown that there is positive correlation (r = 0.91) between the level of lipidic peroxides and the extent to which the tissue is affected. The latter is assessed by studying the serum activity of lactate dehydrogenase. Therefore, these two biologic parameters are shown to be very useful when the study of the development of the diseases is undertaken. PMID- 10822522 TI - The interleukin-1, interleukin-2, interleukin-6 and tumour necrosis factor alpha serological levels in localised and systemic sclerosis. AB - Serological level of interleukin-1 (IL-1), Interleukin-2 (IL-2), Interleukin-6 (IL-6) and tumour necrosis factor (TNF) alpha was investigated in 26 patients with scleroderma, divided into three lots, by the extension and the progress of the disease. Determinations were performed by ELISA in attack and in remission (after treatment with prednison). Normal values: IL-1 (0-5 pg/ml), IL-2 (0-5 pg/ml), IL-6 (5-15 pg/ml), TNF (0-16 pg/ml). Lot A. Results obtained at the first determination showed that IL-1 is elevated in 4 cases (10-15 pg/ml), IL-2 in 5 cases (10-32 pg/ml), IL-6 in 5 cases (15-42 pg/ml) and TNF in 4 cases (18-34 pg/ml). In the second determination IL-1 was increased in 1 case (8 pg/ml), IL-2 in 1 case (9 pg/ml), IL-6 in 2 cases (12 pg/ml) and TNF was normal. Lot B. In the first determination IL-1 was elevated in 5 cases (8-12 pg/ml), IL-2 in 5 cases (10-15 pg/ml), IL-6 in 7 cases (16-20 pg/ml) and TNF was raised in 3 cases (18-25 pg/ml). At the second determination IL-1 showed normal values in all the cases, IL-2 was raised in 2 cases (10 pg/ml), IL-6 in 2 cases (12.15 pg/ml), TNF in 1 case (20 pg/ml). Lot C. In the first determination there were raised values in 4 cases for IL-1 (6-8 pg/ml), 3 cases for IL-2 (10-18 pg/ml), 5 cases for IL-6 (18 20 pg/ml), 2 cases for TNF (20 pg/ml). At the second determination IL-2 was elevated in 1 case (10 pg/ml), IL-6 in 1 case (15 pg/ml). We consider that in scleroderma there is a disturbance of the investigated cytokines due to the activation and involvement of the secretory cells into the pathogenesis of the disease. The increase of the serological levels of IL-1, IL-2, IL-6 and TNF depends on the extension of the lesions and the clinical and biological activity periods of the disease. The absence of the increase of the serological levels does not exclude their activity at the lesional site. PMID- 10822523 TI - Study of islet cell antibodies as predictive marker of type 1 diabetes. AB - The aim of this study was to evidence whether islet cell antibodies (ICAs) (by a standard indirect immunofluorescent technique) could be used to identify subjects at risk for type-1 diabetes. In the present prospective diabetes family study, 23 of 2642 first-degree relatives of patients with type-1 diabetes have developed the disease. The antibody status of these 23 samples was compared with that from 23 carefully matched control. At a mean of 26 years before the onset of diabetes, ICAs were found in 16 patients at the titer 1:60 (versus O control subjects, p < 0.001). The authors consider that ICAs may be interpreted as an immunological change in the islets of first-degree relatives of patients with type-1 diabetes. PMID- 10822524 TI - Considerations on the changes of certain immunologic cells in silicosis. AB - Cell-mediated immunity was estimated in a group of 68 patients divided into three subgroups according to the evolutive stage of disease. It was demonstrated that in silicosis there is a disturbance of immunity. The detection of E rosettes and of activated rosettes was carried out according to the classical method. The mean percentage of total T lymphocytes was found lower in silicosis (62 +/- 2% statistically significant for p < 0.05), compared to the control group (66 +/- 4.6%). The study revealed a decrease of T helper lymphocytes, an increase of T suppressor/cytotoxic lymphocytes and a decrease of the T helper/T suppressor ratio (1.96 +/- 0.13) in silicosis as compared to the control group (2.83 +/- 0.31). The presence of these changes in silicotic patients suggested the participation of cell-mediated immunity in the pathogenesis of disease, the cell mediated immunity being directly responsible for the evolution of silicosis. PMID- 10822525 TI - Antiinflammatory effect of laser therapy in rheumatoid arthritis. AB - The effect of a 940-980 nm length wave laser radiation on rheumatoid arthritis was analyzed in 60 patients: 44 adults and 16 children. Each joint was daily treated by irradiation during 7 minutes for a period of 10 days. The evolution was estimated using an analogic scale with 3 steps for 2 parameters: local pain and local inflammation for each treated joint. The evolution was favourable in both aspects, but the inflammation seemed to be better influenced than the pain. The evolution was better in children than in adults. The inflammations of the interphalangeal, carpometacarpal and shoulder's joints were a better evolution than the knee inflammation. The serological data were not influenced by laser therapy. When relapse never the treated joint was affected again. No adverse or unwanted effects were observed during the treatment excepting a sedation and an increase of night sleep duration. PMID- 10822526 TI - The outpatient management of peripheral diabetic neuropathy. AB - Diabetic neuropathy is a common complication of diabetes. Its commonest form is the bilateral, distal sensorimotor neuropathy and this has been associated with increased risk of disability from foot ulceration, inadvertent injury leading to gangrene as well as to amputation. The economic implications of the treatment of diabetic neuropathy and its consequences are enormous. In spite of this, there is no universally accepted method of treating it and efforts are still underway to find an adequate form of therapy. The following is a review of the outpatient management of peripheral diabetic neuropathy. PMID- 10822527 TI - Total syntheses of (+/-)-preussomerins G and I. AB - [formula: see text] Preussomerins G and I (2 and 3) have been synthesized for the first time. The key reaction in the synthesis is a possibly biomimetic tautomerization reaction depicted in Scheme 3 and the foregoing graphic. The driving force for this interesting rearrangement is primarily derived from the increase in resonance energy associated with converting a naphthalene ring into two isolated benzene rings. PMID- 10822528 TI - A novel anionic condensation, fragmentation, and elimination reaction of bicyclo[2.2.1]heptenone ring systems. AB - [formula: see text] We have identified an unprecedented anionic condensation, fragmentation, and elimination sequence from the coupling of bicyclo[2.2.1] heptenones with aldehydes. This reaction leads to the stereoselective formation of disubstituted five-membered rings which are present in a wide array of bioactive molecules. PMID- 10822529 TI - The development of efficient protocols for the palladium-catalyzed cyclization reactions of secondary amides and carbamates. AB - [formula: see text] With the proper choice of palladium catalyst, ligand, and base, five-, six-, and seven-membered rings are formed efficiently from secondary amide or secondary carbamate precursors, offering significant improvements to currently existing methodology. PMID- 10822530 TI - A unified and quantitative receptor model for the microtubule binding of paclitaxel and epothilone. AB - [formula: see text] Paclitaxel and epothilone represent the two major classes of antimicrotubule agents that promote tubulin polymerization and, presumably, mitotic arrest during cell division. A common minireceptor binding site model at beta-tubulin has been constructed for these structurally divergent compounds. Utilizing 20 amino acids identified in photoaffinity labeling experiments, the 3 D model correlates measured and predicted Ki's with r = 0.99 and rms(delta Gcalc delta Gexp) = 0.2 kcal/mol. In addition, the model predicts the affinity of compounds not used in the training set and explains much of the SAR for the paclitaxel and epothilone families. PMID- 10822531 TI - Unique "cradled barbell" complex between a secondary diammonium ion and bis(m phenylene)-32-crown-10. AB - [formula: see text] The complexation between N,N'-dibenzyl(m-xylylene)diammonium bis(hexafluorophosphate) (2) and bis(m-phenylene)-32-crown-10 (5) was shown to occur in solution by nuclear magnetic resonance with 1:1 stoichiometry and a Ka value of 189 +/- 19 M-1. A crystal structure of 2:5 revealed a unique 1:1 "exo" or "cradled barbell" complex, instead of the expected pseudorotaxane. This unexpected result illustrates that caution be used in interpreting the results from these types of complexes in the solution and "gas" phases on the basis of crystal structures. PMID- 10822532 TI - Enantioselective binding of an 11-cis-locked cyclopropyl retinal. The conformation of retinal in bovine rhodopsin. AB - [formula: see text] The conformation of the retinal chromophore in rhodopsin is central for understanding the visual transduction process. The absolute twist around the 12-s bond of the chromophore in rhodopsin has been determined by studies with 11-cis-locked 11,12-cyclopropylretinal analogues (11S,12R)-2 and (11R,12S)-3, enantioselectively synthesized with the aid of an enzyme. The finding that enantiomer 2 binds to opsin while the other 3 does not defines the absolute sense of twist around the 12-s bond. PMID- 10822533 TI - Enantioselective aldol cyclodehydrations catalyzed by antibody 38C2. AB - [formula: see text] Aldolase antibody 38C2 catalyzes the enantioselective aldol cyclodehydration of 4-substituted-2,6-heptanediones (3) to give enantiomerically enriched 5-substituted-3-methyl-2-cyclohexen-1-ones (4). Yields, enantioselectivities, and product purities are markedly increased compared to the L-proline-catalyzed reactions. PMID- 10822534 TI - Novel strategies to construct the gamma-hydroxy lactone moiety of the CP molecules. Synthesis of the CP-225,917 core skeleton. AB - [formula: see text] The array of challenging structural lineaments embodied in the CP molecules (1 and 2, Figure 1) offers synthetic chemists uncharted realms of exploration and discovery. In this communication, we focus on the chemical hurdies posed by the gamma-hydroxy lactone moiety of these exciting targets. Thus, the examination of the general reactivity of these systems, the development of a novel tandem oxidation sequence to construct the gamma-hydroxy lactone moiety, and the successful construction of the complete polycyclic core of 2 (compound 28, Scheme 5) is enumerated within. PMID- 10822535 TI - On the synthesis of Z-gamma-amino-alpha,beta-unsaturated esters via Ru-catalyzed coupling. AB - [formula: see text] A synthesis of gamma-amino acids and their cyclic derivatives, 3-pyrrolinones, from alpha-amino acids employs a ruthenium-catalyzed Alder ene reaction as a key step in the sequence. Cyclopentadienylruthenium (1,4 cyclooctadiene) chloride catalyzes the addition of gamma-amido-alpha,beta alkynoate esters with monosubstituted alkenes. The major product arises from C-C bond formation at the alpha-carbon of the alkynoate. The reaction exhibits high chemoselectivity. The ruthenium-catalyzed addition occurs in preference to a Diels-Alder reaction. The two new double bonds are created with complete geometrical control. The initial gamma-amido-alpha,beta-unsaturated ester can be cyclized to the pyrrolinones with a tin catalyst. PMID- 10822536 TI - Total synthesis of 3-deoxy-D-manno-2-octulosonic acid (KDO) and 2-deoxy-beta-KDO. AB - [formula: see text] Total syntheses of KDO and 2-deoxy-beta-KDO are reported. The C2-symmetric dienediol 4 was desymmetrized by conversion to its corresponding 1,4 dioxanone 5. Ireland-Claisen rearrangement of 5 provided the 6-vinyldihydropyran 2-carboxylate template 6. Double-Sharpless asymmetric dihydroxylation gave the tetraol 7a, which was converted to KDO and 2-deoxy-beta-KDO using methods similar to those previously reported. This synthetic scheme provides a flexible route to KDO and KDO analogues. PMID- 10822537 TI - Enantioselective synthesis of saframycin A and evaluation of antitumor activity relative to ecteinascidin/saframycin hybrids. AB - [formula: see text] A short synthesis of saframycin A is described which begins with a readily available intermediate previously utilized for the total synthesis of ecteinascidin 743. A key step in this synthesis is the use of 1-fluoro-3,5 dichloropyridinium triflate to oxidize a phenolic ring to a 1,4-benzoquinone unit while simultaneously cleaving a methoxymethyl ether of a different phenolic ring to the corresponding phenol (4-->5). The common intermediate (2) for the synthesis of saframycin A (1) and ecteinascidin 743 also allowed the synthesis of two hybrids of these structures (6 and 7). Whole cell bioassays for antitumor activity using lung, colon, melanoma, and prostate-derived tumor cell lines allowed a clear correlation of structure with biological activity in this series. PMID- 10822538 TI - An enantioselective total synthesis of (+)-geissoschizine. AB - [formula: see text] A concise asymmetric synthesis of the indole alkaloid (+) geissoschizine (1) has been completed. The synthesis features the highly diastereoselective vinylogous Mannich reaction of 3 with 4 to give 5, which is elaborated into the key tetracyclic intermediate 7 in two steps. Following the stereoselective introduction of the ethylidene moiety to give 9, reduction of the lactam and radical decarboxylation via an acyl selenide gave 12, which was converted into (+)-geissoschizine by formylation. The synthesis requires only 11 chemical operations and proceeds in an overall yield of 17%. PMID- 10822539 TI - Synthesis of the angiotensin converting enzyme inhibitor (-)-A58365A via an isomunchnone cycloaddition reaction. AB - [formula: see text] The angiotensin converting enzyme inhibitor (-)-A58365A (1) was synthesized by a process based on the [3 + 2]-cycloaddition reaction of a phenylsulfonyl-substituted isomunchnone intermediate. The starting material for this process was prepared from L-pyroglutamic acid and involved using a diazo phenylsulfonyl-substituted pyrrolidine imide. Treatment of the diazoimide with Rh2(OAc)4 in the presence of methyl vinyl ketone afforded a 3-hydroxy-2-pyridone derivative which was subsequently converted to the ACE inhibitor in six additional steps. PMID- 10822540 TI - Selective lithiation of 2-methyloxazoles. Applications to pivotal bond constructions in the phorboxazole nucleus. AB - [formula: see text] The lithiation of 2-methyloxazoles with alkyllithium and hindered lithium amide bases generally results in the competitive formation of a mixture of 5-lithio- and 2-(lithiomethyl)oxazole isomers. Herein a synthetically useful lithiation method which allows for the selective formation of 2 (lithiomethyl)oxazole is described. Diethylamine has been found to be a kinetically competent proton source that will mediate the equilibration of the kinetically formed 5-lithiooxazole to its more stable 2-(lithiomethyl)oxazole counterpart. Application of this metalation strategy with lithium diethylamide to two important bond constructions relevant to a projected phorboxazole synthesis is presented. PMID- 10822541 TI - 3-(Diethoxyphosphoryloxy)-1,2,3-benzotriazin-4(3H)-one (DEPBT): a new coupling reagent with remarkable resistance to racemization. AB - [formula: see text] The new crystalline phosphate reagent 3 (diethoxyphosphoryloxy)-1,2,3-benzotriazin-4(3H)-one (DEPBT) mediates amide bond formation with a remarkable resistance to racemization. Comparative racemization studies were carried out and DEPBT proved to be superior to typical phosphonium and uronium coupling reagents. DEPBT is easily prepared and is exceedingly stable with a shelf life of months at room temperature. The advantageous properties of DEPBT render it a useful and unique addition to the arsenal of coupling reagents. PMID- 10822542 TI - Studies on the synthesis of tedanolide: synthesis of the C(5)-C(21) segment via a highly stereoselective fragment assembly aldol reaction of a chiral beta,gamma unsaturated methyl ketone. AB - [formula: see text] A highly diastereoselective synthesis of 3, corresponding to the C(5)-C(21) segment of tedanolide, has been accomplished by a route utilizing the aldol reaction of aldehyde 4 and the beta,gamma-unsaturated methyl ketone 5. PMID- 10822543 TI - Synthesis of a reaction intermediate analogue of biotin-dependent carboxylases via a selective derivatization of biotin. AB - [formula: see text] An efficient and practical synthesis of 1, a unique reaction intermediate analogue of biotin-dependent carboxylases, is described. The synthesis features a selective acylation of the 1'-N of biotin. Target 1 inhibits the activity of the biotin carboxylase component of acetyl CoA carboxylase. It is the first known biotin-derived inhibitor of biotin carboxylase and should promote new kinetic and structural studies of the biotin-dependent carboxylases. PMID- 10822544 TI - Can lone pairs bind to a pi system? The water...hexafluorobenzene interaction. AB - [formula: see text] Ab initio calculations reveal a significant binding interaction between water and hexafluorobenzene in a geometry that points the oxygen lone pairs directly into the face of the pi system. The geometry is as anticipated from electrostatic arguments emphasizing the substantial quadrupole moment of the aromatic. A second, off-axis geometry is also found which is also consistent with a substantial electrostatic interaction. PMID- 10822545 TI - Glycosyl transfer to nitrogen via cycloaddition. AB - [formula: see text] This letter describes the reduction to practice of a novel concept for functionalization of the anomeric carbon of carbohydrates with a nitrogen substituent. Thus, bisheterodienes with a thiono sulfur terminus and a sulfonylimine terminus are shown to undergo cycloaddition smoothly and stereoselectively to three different glycals. PMID- 10822546 TI - Antihydrophobic cosolvent effects in organic displacement reactions. AB - [formula: see text] Rates of reactions in water can be modified by the presence of antihydrophobic cosolvents such as ethanol and DMSO, which lower the energies of nonpolar surfaces. The rate effects reflect changes both in the solvation of nonpolar surfaces and also in the solvation of polar groups. The effects have been sorted out for some displacement reactions, revealing the geometry of an interesting branching reaction whose two paths show different antihydrophobic effects. PMID- 10822547 TI - Solid-phase SN2 macrocyclization reactions to form beta-turn mimics. AB - [formula: see text] Efficient solid-phase SN2 macrocyclization reactions were sought to facilitate preparations of focused libraries of beta-turn mimetics. A very efficient, but undesired, cyclization reaction to give five-membered ring lactams 4 was identified in attempts to use O-nucleophiles. Subsequent studies focused exclusively on S-nucleophiles. These reactions gave the desired macrocyclization products 1 in high purities and good overall yields. Conformational analyses of illustrative macrocyclization products 1 via NMR, CD, and molecular simulations showed that they seem to sample both type I and type II beta-turn conformations in solution. CD studies indicate a curious relationship between the preferred conformation and the amino acids encapsulated in the macrocycles. PMID- 10822548 TI - Biosynthesis of desosamine: construction of a new macrolide carrying a genetically designed sugar moiety. AB - [formula: see text] The appended sugars in macrolide antibiotics are indispensable to the biological activities of these important drugs. In an effort to generate a set of novel macrolide derivatives, we have created a new analogue of methymycin and neomethymycin, antibiotics produced by Streptomyces venezuelae. This analogue 15 carrying a different sugar, D-quinovose, instead of D desosamine, was constructed by taking advantage of targeted gene deletion combined with a specific pathway-independent C-3 reduction capability of the wild type S. venezuelae. PMID- 10822549 TI - Role of aggregates in Claisen acylation reactions of imidazole, pyrazole, and thioesters with lithium enolates in THF. AB - [formula: see text] Although phenyl esters react with both monomers and dimers or tetramers of two lithium enolates in THF, the reactions of phenyl thiobenzoates are relatively much faster with the monomers. Similarly, imidazole esters react primarily with the monomers but pyrazole esters react with monomers and aggregates. The results are rationalized by a mechanism in which coordination with two lithium cations within an enolate aggregate is required for the reaction of aggregates to compete with monomers. PMID- 10822550 TI - N,O-Bis(trifluoroacetyl)hydroxylamine as a useful electrophilic nitrogen source: catalytic synthesis of N-(trifluoroacetyl)sulfilimines. AB - [formula: see text] In the presence of catalytic quantites of Cu(OTf)2 the novel hydroxamic acid anhydride salt functions competently in the trifluoroacetamidation of sulfides to afford N-(trifluoroacetyl)sulfilimines. The salient features of this salt include its ease of synthesis from the inexpensive, commercially available starting materials trifluoroacetic anhydride and hydroxylamine hydrochloride. PMID- 10822551 TI - Heterolytic cleavage of a beta-phosphatoxyalkyl radical resulting in phosphate migration or radical cation formation as a function of solvent polarity. AB - [formula: see text] The 2-(diethylphosphatoxy)-2-(p-methoxyphenyl)-1,1 dimethylethyl radical (1) reacted to give the benzylic radical product from phosphate migration or a radical cation (or a mixture of the two) as a function of solvent. Smooth acceleration in rates of reactions of 1 in solvents of increasing polarity and consistent entropies of activation indicate that radical 1 reacts by common mechanism irrespective of the final products formed, specifically by initial heterolysis to a radical cation-phosphate anion pair. PMID- 10822552 TI - Enantioselective synthesis of alpha-amino nitriles from N-benzhydryl imines and HCN with a chiral bicyclic guanidine as catalyst. AB - [formula: see text] A novel catalytic enantioselective Strecker synthesis of chiral alpha-amino nitriles and alpha-amino acids is described and analyzed with regard to the possible mechanistic basis for stereoselectivity. Key features of the enantioselective process include (1) the use of the chiral bicyclic guanidine 1 as catalyst and (2) the use of the N-benzhydryl substituent on the imine substrate. PMID- 10822553 TI - Novel [1,5] sigmatropic rearrangements of cyclohexadienones generated from Fischer carbene complexes. A new strategy for installing the C-20 angular ethyl group in Aspidospermidine alkaloids. AB - [formula: see text] We report here the first examples of a [1,5] sigmatropic rearrangement in a 4a-alkyl-4a-hydrocarbazol-4-one to yield a 3-alkylcarbazol-4 one with a re-aromatized indole nucleus. The reaction of 1-methyl-3-substituted indole-2-carbene complexes 1 with terminal alkynes yields 3,4a-dialkyl-1-methoxy 9-methylcarbazol-4-ones 2. These 4a-substituted carbazolones thermally rearrange to cleanly give the more highly aromatic 3,3-dialkyl-1-methoxy-9-methylcarbazol-4 ones 3. This reaction provides a convenient entry to the Aspidosperma family of alkaloids, which contain a 3,3-disubstituted carbazole nucleus. PMID- 10822554 TI - Palladium-catalyzed highly diastereoselective cyclic carbopalladation carbonylative esterification tandem reaction of iododienes and iodoarylalkenes. AB - [formula: see text] Pd-catalyzed reaction of iododienes and iodoarylalkenes represented by 1, 8, and 10 under 1 atm of CO and a small amount of O2 in the presence of a base, e.g., NEt3, as well as MeOH and H2O in DMF can undergo a highly diastereoselective cyclic carbopalladation-carbonylative esterification tandem process (Type II C-Pd process) to give in high yields the corresponding ester-containing cyclization products, e.g., 2, 9, and 11, in as high as 98% diastereoselectivity. PMID- 10822555 TI - Synthesis of a C(1)-C(14)-containing fragment of callipeltoside A. AB - [formula: see text] A C(1)-C(14)-containing fragment of callipeltoside A (1, Scheme 1) was synthesized efficiently via a dianion aldol coupling reaction between aldehyde 2 and ketoester 3. A surprising lack of reactivity between the alkenes in 13 and the Grubbs initiator 15 was encountered. An equally surprising rate acceleration of the reaction between 15 and allylic alcohols (alk-1-en-3 ols) as well as their subsequent cleavage to methyl ketones was discovered. In situ 1H NMR analysis has proven to be a very useful tool for monitoring RCM reactions of complex substrates such as 13. PMID- 10822556 TI - A convenient method for synthesis of enantiomerically enriched methylphenidate from N-methoxycarbonylpiperidine. AB - [formula: see text] This report describes a new method to prepare optically active methylphenidate starting from piperidine. The method consists of a transformation of N-methoxycarbonylpiperidine to the corresponding alpha methoxylated carbamate I by utilizing electrochemical oxidation followed by the coupling reaction with optically active Evans imides II to produce optically active methylphenidate derivatives III with high stereoselectivities, threo (2R,2'R)-Methylphenidate (IV; Ar = Ph; Ritalin) was easily prepared from III in three steps. PMID- 10822557 TI - Reduction of azides to primary amines in substrates bearing labile ester functionality. Synthesis of a PEG-solubilized, "Y"-shaped iminodiacetic acid reagent for preparation of folate-tethered drugs. AB - [formula: see text] Anhydride 3 is a useful reagent for the synthesis of triply linked drug conjugates. Examples using paclitaxel are provided. Conversion of the azido moiety to a primary amine in the presence of substrates bearing labile ester functionality requires the use of tin/mercaptan reducing system which includes methanol exchange equilibration to effect nitrogen-tin bond scission. PMID- 10822558 TI - The pattern of fluorine substitution affects binding affinity in a small library of fluoroaromatic inhibitors for carbonic anhydrase. AB - [formula: see text] A library of fluoroaromatic inhibitors of carbonic anhydrase has been found to bind in a manner dependent on both hydrophobicity and the pattern of substitution of the fluoroaromatic ring. All of the compounds in the library bind to the protein with Kd < 3 nM. We have inferred two distinct binding modes from our data, which suggest two types of interactions that should be considered when designing fluorinated drugs. PMID- 10822559 TI - A caged hydrophobic inhibitor of carbonic anhydrase II. AB - [formula: see text] A tight-binding, hydrophobic inhibitor of carbonic anhydrase II has been masked with a water-solubilizing, photolabile group derived from o nitrophenylglycine. This caged inhibitor represents our first effort at the site specific delivery of prodrugs that can be activated by light. Via this approach, we have begun to address the problems of water insolubility and systemic side effects on administration of tight-binding inhibitors of carbonic anhydrase. PMID- 10822560 TI - IMDA/retro-Mannich approach to cis-perhydroquinoline Lycopodium alkaloids: asymmetric synthesis of (+)-luciduline. AB - [formula: see text] The first chiral auxiliary mediated asymmetric synthesis of the naturally occurring Lycopodium alkaloid (+)-luciduline has been accomplished. Key steps include an IMDA reaction of a chiral dihydropyridine, a subsequent retro-Mannich ring opening, and a novel cationic reductive cyclization reaction. PMID- 10822561 TI - Heterocyclic HIV-1 protease inhibitors. AB - [formula: see text] A series of simple heterocyclic HIV-1 protease inhibitors were developed on the basis of size, shape, and electronic complementarity to the active site of the enzyme. The C2-symmetric heterocycles do not contain a transition-state isostere nor are they active site directed irreversible inhibitors; thus, they represent the success of a new design strategy. The first generation heterocycles inhibit the protease in the micromolar range, whereas control compounds show no bioactivity at the same concentrations. PMID- 10822562 TI - Single- and double-proton-transfer in the aggregate between cytosine and guaninediazonium ion. AB - [formula: see text] The structure of "guaninediazonium ion" in its aggregate with cytosine has been explored with ab initio and density functional methods. The hydrogen-bonded aggregate between cytosine and guaninediazonium ion, 1, is a stable minimum, 3. While the isolated enol tautomer of guaninediazonium ion, 2, is significantly more stable than 1, the tautomeric aggregate 4 that results from double-proton-transfer in 3 is almost isoenergetic with 3. Most interesting and entirely unexpected is the finding that neither 3 nor 4 is predicted to be the thermodynamically predominant structure. Instead, single-proton-transfer to cytosine results in the most stable cytosinium-guaninediazo complex, 5. PMID- 10822563 TI - An enyne metathesis/(4 + 2)-dimerization route to (+/-)-differolide. AB - [formula: see text] A concise total synthesis of (+/-)-differolide (1) has been achieved. 2-Vinylbutenolide (2) was prepared by enyne metathesis of allyl propynoate (3) using the Grubbs initiator 4. This reaction was examined by 1H NMR spectroscopy, which led to the hypothesis that low concentration of ruthenium species and high concentration of enyne substrate would be advantageous. Accordingly, slow addition of 4 to solutions of enyne 3 was found to be beneficial. Spontaneous dimerization of 2 gave (+/-)-differolide (1) and an isomer. PMID- 10822564 TI - Catalytic asymmetric allylation reactions using BITIP catalysis and 2-substituted allylstannanes as surrogates for beta-keto ester dianions. AB - [formula: see text] Catalytic asymmetric allylation (CAA) reactions using the indicated allylstannane and the BITIP catalysts previously described by us give high yields and enantioselectivities in additions to aldehydes. The products are convertible to beta-keto esters by oxidative cleavage of the olefin. These reactions thus provide a useful catalytic enantioselective method for chain extension with introduction of a versatile four-carbon unit. PMID- 10822565 TI - Efficient enantioselective synthesis of sertraline, a potent antidepressant, via a novel intramolecular nucleophilic addition to imine. AB - [formula: see text] An efficient enantioselective synthesis of sertraline, an antidepressant, utilizing anionic imine ring closure is described. PMID- 10822566 TI - Synthesis of beta-hydroxy-beta-(fluoronitrophenyl)alanines: vital components in the assembly of biologically active cyclic peptides. AB - [formula: see text] Numerous biologically active cyclic peptides, such as the antibiotic vancomycin, contain amino acid residues connected through side-chain biaryl or aryl-alkyl ether linkages. Nucleophilic aromatic substitution reactions have recently been shown to provide a general method for the formation of such ether linkages, and consequently the synthesis of functionalized fluoronitro substituted aromatic amino acids is of great interest. Herein, a method for the stereospecific synthesis of 3-fluoro-4-nitro- and 4-fluoro-3-nitro-threo-beta hydroxyphenylalanine is described. PMID- 10822567 TI - Synthesis of four diastereomeric 3,5-dialkoxy-2,4-dimethylalkanals by a simple extension of the non-aldol aldol process to bis(propionates). AB - [formula: see text] The synthesis of all four diastereomers of bis(propionates), 3,5-dialkoxy-2,4-dimethylalkanals, by non-aldol aldol chemistry is described. The epoxy alcohols (3, 4) were converted into the mesylates (9, 11) which were cleanly rearranged to the desired 3,5-bis(oxygenated)-2,4-dimethylalkanals (10, 12) in high yield. The epoxy mesylates (13, 16) gave the desired products (14, 17) in good yield on treatment with TMSOTf and a hindered base. PMID- 10822568 TI - Site-specific cross-linking of nucleic acids using the abasic site. AB - [formula: see text] An efficient site-specific cross-linking reaction between two carbohydrate residues present in two complementary DNA sequences is described. One oligodeoxynucleotide, 5'd(GGCTGA*CTGCG)3', carries an amino nucleophile tethered to the 2'-hydroxyl of an adenosine residue (A*). The target electrophile is an abasic site generated in the complementary sequence, 5'd(CGCAGDCAGCC)3' (D represents the deoxyribose). The cross-linking reaction was carried out by a reductive amination reaction in > 95% yield. PMID- 10822569 TI - Novel synthesis of the C1-C15 polyether domain of the thyrsiferol and venustatriol natural products. AB - [formula: see text] A convergent construction of the C1-C15 domain of the thyrsiferol-related natural products has been developed. This involved the separate construction of C1-C7 and C8-C15 fragments, their organochromic-mediated coupling, and subsequent reductive closure of the B ring. This synthetic A-B-C ring construct will be useful for the total synthesis of the biologically active polyether squalenoid natural products, as well as their non-natural analogues. PMID- 10822570 TI - 13C and 2H kinetic isotope effects and the mechanism of bromination of 1-pentene under synthetic conditions. AB - [formula: see text] The 13C and 2H kinetic isotope effects for the bromination of 1-pentene with Br2 in CCl4 were determined and interpreted with the aid of calculationally predicted isotope effects. The isotope effects observed are consistent with rate-limiting bromonium ion formation and do not fit with either rate-limiting production of a pi complex or reaction of a reversibly formed bromonium ion. This rules out some of the mechanistic complexities suggested for other brominations, though the identity of the brominating reagent(s) under these synthetic conditions remains uncertain. PMID- 10822571 TI - Total synthesis of (+/-)-methyl rishirilide B. AB - [formula: see text] A regio- and stereospecific total synthesis of (+/-)-methyl rishirilide B (2b), and (alpha)2-macroglobulin inhibitor, is described. A key feature of the synthetic plan was regiospecific construction of a hydroanthracenone intermediate through condensation of a phenylsulfonyl isobenzofuranone with a functionalized 2-cyclohexen-1-one. Introduction of the vicinal trans-hydroxyl groups in the densely functionalized A-ring was accomplished via a novel one-pot procedure that involved oxidation of enolate anions with the Davis reagent. PMID- 10822572 TI - Isolation, structure elucidation, and synthesis of bisgersolanolide, a novel heptacyclic bis-diterpenoid from the gorgonian octocoral Pseudopterogorgia bipinnata. AB - [formula: see text] A chemical study of the hexane extracts of the Caribbean gorgonian Pseudopterogorgia bipinnata (Verrill) collected in San Andres Island, Colombia, has led to the isolation of an unprecedented heptacyclic C40 bis diterpenoid, bisgersolanolide (1). The structure of this novel secondary metabolite, which was established by spectroscopic studies that included 2D NMR correlation methods, IR, UV, and accurate mass measurements, was confirmed by synthesis. PMID- 10822573 TI - Synthesis of 1,2,3,7,8,9-hexahydrodipyrrins and secocorrins: important precursors for the construction of corrins. AB - [formula: see text] Hexahydrodipyrrins 15 (X=H2) have been prepared by two routes: Pd(0)-initiated coupling/cyclization of triflates 11 and alkyne amines 12 and reduction of semicorrins 26 (X=O). PMID- 10822574 TI - Nonstabilized N-unsubstituted azomethine ylides: a synthesis of indolizidine 239CD. AB - [formula: see text] Treatment of a (2-azaallyl)stannane with HF.pyridine generated a nonstabilized N-unsubstituted azomethine ylide, which was found to undergo an efficient and stereoselective dipolar cycloaddition with phenyl vinyl sulfone to produce a trans-2,5-dialkylpyrrolidine that was further transformed into the dendrobatid alkaloid indolizidine 239CD. PMID- 10822575 TI - Asymmetric epoxy cyclohexenyl sulfones: readily accessible progenitors of stereo defined six-carbon arrays. AB - [formula: see text] Enantiopure epoxy vinyl sulfones serve as highly effective substrates for a variety of stereo- and regiospecific oxidation and nucleophilic functionalization reactions. These materials can be easily transformed to cyclic and acyclic six-carbon segments. Nucleophilic epoxidation of 3a,b followed by palladium[0] catalysis enables access to differentially protected arene diols 21 and 22. PMID- 10822576 TI - Synthesis of NBD-alpha-galactosylceramide and its immunologic properties. AB - [formula: see text] A representative alpha-galactosylceramide (alpha-GalCer), KRN7000, can activate NKT cells through CD1d molecules, which play an essential role in the generation of the strong antitumor activity of KRN7000. Our previous study has demonstrated that alpha-GalCer binds directly to CD1d molecules. However, it is controversial whether CD1d binds alpha-GalCer in endosomal compartments. To address this question, we synthesized NBD-alpha-GalCer, which has strong fluorescent properties. We found that the NBD-alpha-GalCer has immunostimulatory activity that is stronger than that of KRN7000. PMID- 10822577 TI - Facile preparation of allenic hydroxyketones via rearrangement of propargylic alcohols. AB - [formula: see text] Treatment of tertiary propargylic alcohols 13 with 3-diazo-2 butanone 6 and catalytic dirhodium tetraacetate in benzene gave good yields of the diastereomeric allenic hydroxyketones 14, with, in some cases, good diastereocontrol. These products are presumably formed via the [2,3]-sigmatropic rearrangement of an alpha-propargyloxy enol derivative. This reaction has been extended to the preparation of homoallylic hydroxyketones from allylic alcohols by reaction with 6 and the rhodium catalyst. PMID- 10822578 TI - Rhodium carbenoid-initiated Claisen rearrangement: scope and mechanistic observations. AB - [formula: see text] It has been shown that alpha-diazoketones react with allylic alcohols in the presence of Rh(II) catalysts to furnish intermediate enols which subsequently undergo Claisen rearrangement to alpha-hydroxyketones. Herein we report (1) studies into the mechanism of this transformation which establish that Claisen rearrangement is neither rhodium- nor acid-catalyzed but a reaction intrinsic to the intermediate enols that proceeds at a rate governed by enol substituents (R3, R4, R5) and (2) the reaction of alpha-diazoketones with propargylic alcohols and preliminary investigations into its scope and mechanism. PMID- 10822579 TI - A new tandem route to angular tetraquinanes. Synthesis of the Waihoensene ring system. AB - [formula: see text] This paper describes a new tandem reaction sequence leading to angularly fused polyquinanes from squaric acid-derived bicyclo[6.3.0] undecadienediones. Such compounds undergo a dual Michael addition. The enolate form in the first intermolecular addition undergoes the second intramolecular transannular addition to give the angular polyquinanes. A particularly interesting example is a catalytic transformation of cis-13 methylyricyclo[10.3.0.0]pentadeca-4(5),12(13)-diene-3 ,14-dione to (3R*,3aS*,5aR*,9aR*,11aR*)-3-methyl-1,2,3,5,5a,6 ,7,10,11,11a-decahydro-4H- pentaleno[6a,1-c]indene-2,10-dione, a compound having the tetracyclic ring system found in the natural product waihoensene. The mechanism and synthetic scope of these reactions are discussed. PMID- 10822580 TI - A highly enantioselective and diastereoselective synthesis of cyclobutanes via boronic esters. AB - [formula: see text] Deprotonation of enantiopure (R,R)-1,2-dicyclohexyl-1,2 ethanediol 1-chloro-4-cyanobutylboronates 5 with LDA followed by treatment with anhydrous magnesium bromide yields (R)-(trans-2-cyanocyclobutyl)boronic esters 7 in high diastereomeric and enantiomeric purity. No cyclobutane formation has been observed in the absence of at least a catalytic amount of magnesium halide. PMID- 10822581 TI - Modular nucleic acid surrogates. Solid phase synthesis of alpha-helical peptide nucleic acids (alpha PNAs). AB - [formula: see text] The synthesis and characterization of prototype alpha-helical peptide nucleic acid (alpha PNA) modules 1-3 as well as disulfide dimers 4 and 5 are reported. These molecules combine an alpha-helical peptidyl scaffold with well-defined nucleobase molecular recognition patterns and could serve as a basis for novel antisense and/or antigene agents. Structure assignments for these alpha PNAs were supported by MALDI-TOF mass spectrometry, and the alpha-helical nature of 4 in water was confirmed by circular dichroism (CD) spectroscopy. PMID- 10822583 TI - Predictions of geometries and multiplicities of the manganese-oxo intermediates in the Jacobsen epoxidation. AB - [formula: see text] The geometries and multiplicities of models of the manganese(III)-salen catalyst and the manganese(V)-oxo Intermediate in the Jacobsen epoxidation were explored with density functional theory (Becke3LYP). Mn(III) complexes are quintet ground states, while ligands influence whether quintet, triplet, or singlet states are lowest in energy for Mn(V)-oxo complexes. Geometries and multiplicities and their implications for stereoselectivity are described. PMID- 10822582 TI - A versatile preparation of alpha,beta-unsaturated lactones from homoallylic alcohols. AB - [formula: see text] A new method for the synthesis of alpha,beta-unsaturated lactones from beta-acetoxy aldehydes by reaction with the lithium enolate of methyl acetate was developed. The reaction is relatively insensitive to structural changes in the aldehyde substrates. The process was extended to the synthesis of five-ring lactones from alpha-acetoxy aldehydes. Experimental evidence regarding the mechanism of this one-pot transformation was obtained. The observations are consistent with a pathway involving an initial aldol condensation with subsequent acyl migration, lactonization, and beta-elimination and not an enolate equilibration-aldol mechanism. PMID- 10822584 TI - Investigation of the stability of thiosialosides toward hydrolysis by sialidases using NMR spectroscopy. AB - [formula: see text] 1H NMR spectroscopy has been used to investigate whether the alpha(2-->6)-linked thiosialoside 3 and the alpha(2-->3)-linked thiosialoside 9 are hydrolyzed in the presence of Vibrio cholerae sialidase. Similarly, the hydrolysis of the O-ketosides Neu5Ac-2-O-alpha-(2-->3)-Gal beta Me (4) and the alpha-(2-->6)-sialyllactoside 7, representing natural alpha(2-->3)- and alpha(2- >6)-linked sialosides, respectively, was investigated. The results of the 1H NMR experiments clearly demonstrate that the thiosialosides are not hydrolyzed by Vibrio cholerae sialidase. As expected, the O-sialosides are hydrolyzed to give N acetyl-alpha-D-neuraminic acid as the first product of substrate cleavage. PMID- 10822585 TI - Abbreviated synthesis of the C3-C14 (substituted 1,7-dioxaspiro[5.5]undec-3-ene) system of okadaic acid. AB - [formula: see text] Described is a novel, concise, and flexible synthesis of the C3-C14 portion of okadaic acid. A substituted valerolactone (C3-C8) was prepared in three steps and alpha-hydroxylated using Davis' oxaziridine. Conjugate addition of dimethylcuprate upon ynones derived from the C3-C8 lactones followed by intramolecular ketalization provided the C3-C14 fragment and revealed a significant role of the C7 alpha'-ketone substituent upon the efficiency of spiroketalization. PMID- 10822586 TI - Bilosespens A and B: two novel cytotoxic sesterpenes from the marine sponge Dysidea cinerea. AB - [formula: see text] Two novel sesterpenes, bilosespens A and B (1 and 2) were isolated from the Red Sea sponge Dysidea cinerea collected in the Dahlak archipelago, Eritrea. The structure of the mixture of the two inseparable compounds was established by spectroscopic analysis, mainly by 1D and 2D NMR measurements. The mixture of bilosespens A and B is cytotoxic to a few human cancer cells. PMID- 10822587 TI - A new, lipophilic p-alkoxybenzyl ether protecting group and its use in the synthesis of a disaccharide. AB - [formula: see text] In contrast to major advances in the chemical synthesis of oligosaccharides, the methods of purification of the intermediates are essentially the same as they were decades ago. Here, the synthesis of p (dodecyloxy)benzyl chloride is described and it is demonstrated that the new p (dodecyloxy)benzyl ether protecting group can render a protected disaccharide sufficiently lipophilic for selective adsorption on C18 silica, thus sidestepping the expensive silica gel chromatography traditionally used for the isolation of protected oligosaccharides. PMID- 10822588 TI - Carbometalations of simple alkenes with allyldibromoborane. AB - [formula: see text] The allylboration of alkenes with allyldibromoborane is described. Utilizing an improved methodology for the synthesis of allyldibromoborane, 1,3-dienes, styrene, and even isolated alkenes could be carbometalated in high yield regio- and stereospecifically at 0 degree C. This high reactivity of allyldibromoborane significantly expands the scope of carbometalations of alkenes. PMID- 10822589 TI - 1,7-Asymmetric induction in a nitrogen ring expansion process facilitated by in situ tethering. AB - [formula: see text] There are only a few methods for the asymmetric ring expansion of prochiral ketones. Symmetrically substituted cyclohexanones can be converted to the corresponding ring-expanded caprolactam with excellent 1,7 diastereoselectivity (> or = 93% ds) and yields (> or = 86%), using a chiral hydroxy azide-mediated Schmidt reaction. PMID- 10822590 TI - A novel chromium(0)-promoted four-component cycloaddition reaction. AB - [formula: see text] Reaction of (eta 6-thiepin-1,1-dioxide)tricarbonylchromium(0) with excess terminal alkynes under photoactivation conditions affords novel pentacyclic adducts formally derived from a sequential [6 pi + 2 pi]/[6 pi + 2 pi]/[2 sigma + 2 pi] cycloaddition process. PMID- 10822591 TI - Synthesis and NMR characterization of a novel class of thienomorphinans. AB - [formula: see text] Synthesis of four novel thieno derivatives 4-7 featuring the codeine skeletal backbone is reported. Characterization by 1H and 13C NMR is also discussed, along with binding profile for opioid receptors. PMID- 10822592 TI - A new approach to phorbol by [4 + 3] oxyallyl cycloaddition and intramolecular Heck reaction. AB - [formula: see text] A new synthetic strategy for a functionalized tricyclic core of phorbol has been developed by means of a [4 + 3] oxyallyl cycloaddition and subsequent intramolecular Heck reaction. The [4 + 3] oxyallyl cycloadduct 7 was chosen as the B-ring precursor of phorbol. Subsequent elaboration took advantage of its well-defined diastereofacial bias to afford the tricycle 5. This method should be of general value in the construction of 6,7- or 5,7-fused bicyclic systems. PMID- 10822593 TI - Novel antimycobacterial benzoxazole alkaloids, from the west Indian Sea whip Pseudopterogorgia elisabethae. AB - [formula: see text] Our screening for marine natural products with anti tuberculosis activity from the West Indian gorgonian coral Pseudopterogorgia elisabethae resulted in the isolation of two active diterpenoid alkaloids, pseudopteroxazole (1) and seco-pseudopteroxazole (2). Their structures were elucidated by NMR spectral analysis, including a variety of two-dimensional techniques. Compounds 1 and 2 are previously undescribed diterpenoids containing the uncommon benzoxazole moiety. Biological screening studies indicated that pseudopteroxazole (1) is a potent growth inhibitor of Mycobacterium tuberculosis H37Rv, while seco-pseudopteroxazole (2) shows moderate to strong inhibitorial activity. PMID- 10822594 TI - Quantum dynamics of the femtosecond photoisomerization of retinal in bacteriorhodopsin. AB - The membrane protein bacteriorhodopsin contains all-trans-retinal in a binding site lined by amino acid side groups and water molecules that guide the photodynamics of retinal. Upon absorption of light, retinal undergoes a subpicosecond all-trans-->13-cis phototransformation involving torsion around a double bond. The main reaction product triggers later events in the protein that induce pumping of a proton through bacteriorhodopsin. Quantum-chemical calculations suggest that three coupled electronic states, the ground state and two closely lying excited states, are involved in the motion along the torsional reaction coordinate phi. The evolution of the protein-retinal system on these three electronic surfaces has been modelled using the multiple spawning method for non-adiabatic dynamics. We find that, although most of the population transfer occurs on a timescale of 300 fs, some population transfer occurs on a longer timescale, occasionally extending well beyond 1 ps. PMID- 10822595 TI - Prediction of the mechanisms of enzyme-catalysed reactions using hybrid quantum mechanical/molecular mechanical methods. AB - The use of hybrid methods, involving both quantum mechanics and molecular mechanics, to model the mechanism of enzyme-catalysed reactions, is discussed. Two alternative approaches to treating the electrostatic interactions between the quantum mechanical and molecular mechanical regions are studied, involving either the inclusion of this term in the electronic Hamiltonian (QM/MM), or evaluating it purely classically (MO + MM). In the latter scheme, possible problems of using force fields that are standard for macromolecular modelling are identified. The use of QM/MM schemes to investigate the mechanism of the enzymes thymidine phosphorylase (ThdPase) and protein tyrosine phosphatase (PTP) is described. For both systems, transition states have been identified using a PM3 Hamiltonian. For ThdPase, concerted motion of the enzyme during the course of the reaction is suggested and, for PTP, a two-step dephosphorylation reaction is indicated, both with quite low barriers. PMID- 10822596 TI - Energy landscapes of biomolecular adhesion and receptor anchoring at interfaces explored with dynamic force spectroscopy. AB - Beyond covalent connections within protein and lipid molecules, weak noncovalent interactions between large molecules govern properties of cellular structure and interfacial adhesion in biology. These bonds and structures have limited lifetimes and so will fail under any level of force if pulled on for the right length of time. As such, the strength of interaction is the level of force most likely to disrupt a bond on a particular time scale. For instance, strength is zero on time scales longer than the natural lifetime for spontaneous dissociation. On the other hand, if driven to unbind or change structure on time scales shorter than needed for diffusive relaxation, strength will reach an adiabatic limit set by the maximum gradient in a potential of mean force. Over the enormous span of time scales between spontaneous dissociation and adiabatic detachment, theory predicts that bond breakage under steadily rising force occurs most frequently at a force determined by the rate of loading. Moreover, the continuous plot (spectrum) of strength expressed on a scale of loge(loading rate) provides a map of the prominent barriers traversed in the energy landscape along the force-driven pathway and reveals the differences in energy between barriers. Illustrated with results from recent laboratory measurements, dynamic strength spectra provide a new view into the inner complexity of receptor-ligand interactions and receptor lipid anchoring. PMID- 10822597 TI - Collective membrane motions of high and low amplitude, studied by dynamic light scattering and micro-interferometry. AB - Undulations of lipid bilayers were experimentally studied for the two limiting cases of high and weak lateral tension using two well established model systems: freely suspended planar lipid bilayers, so-called black lipid membranes (BLM) for high-tension studies and large unilamellar vesicles (LUV) for measurements at weak tension. This variation in tension results in changes of undulation amplitudes from several hundred nm (LUV) down to 1 nm (BLM), thus requiring different physical methods for their detection. We have employed microinterferometric techniques (RICM) for studying the regime of weak tension and dynamic light scattering (DLS) for that of high tension. The dedicated DLS set-up allowed the measurements of undulations over a wide wave vector range of 250 < q/cm-1 < 35,000 cm-1. This enabled the observation of collective membrane modes in two regimes, the oscillating one at low q and the overdamped regime at high q. The transition between both regimes at the bifurcation point is rather abrupt and depends on the lateral tension of the bilayer, as is demonstrated by comparing the dispersion curves of pure lipid and of lipid-cholestrol BLMs over the same q-range. The DLS measurements allowed a critical test of a hydrodynamic theory of the dispersion behaviour of membrane collective modes under tension. The DLS measurements are compared with RICM results of undulatory excitations of giant vesicles weakly adhering to substrates in the 10(-6)-2.5 x 10(-7) m wavelength regime and at low frequencies (0.1-25 Hz). Experimental evidence for the strong decrease in the relaxation rate by the hydrodynamic coupling of the membrane with the wall is established. PMID- 10822598 TI - Trapping of short-lived intermediates in phospholipid phase transitions: the L* alpha phase. AB - Time-resolved small-angle X-ray diffraction of liquid-crystalline phospholipid water systems under temperature or pressure jump conditions has demonstrated the existence of an ordered, intermediate L alpha phase, with a sub-second lifetime, designated as the L* alpha-phase. The lamellar repeat spacing is, universally, 0.3 nm smaller than that of the parent phase, irrespective of the lipid composition and of the jump conditions, provided that the jump leads to a net volume expansion of the phase. The presence of salts, most notably LiCl, leads to a prolongation of the lifetime. The results suggest a non-monotonic potential function for the interbilayer water thickness. PMID- 10822599 TI - Sensing isothermal changes in the lateral pressure in model membranes using di pyrenyl phosphatidylcholine. AB - In this work we present data from a homologous series of di-pyrenyl phosphatidylcholine (dipyPC) probes which can sense lateral pressure variations in the chain region of the amphiphilic membrane (lateral pressures are tangential to the interface). The dipyPC has pyrene moieties attached to the ends of equal length acyl chains on a phosphatidylcholine molecule. Ultraviolet stimulation produces both monomer and excimer fluorescence from pyrene. At low dilutions of dipyPC in model membranes the excimer signal is entirely intra-molecular and since it depends on the frequency with which the pyrene moieties are brought into close proximity, the relative intensity of the excimer to monomer signal, eta, is a measure of the pressure. We synthesised or purchased dipyPC probes with the pyrene moieties attached to acyl chains having 4, 6, 8 and 10 carbon atoms and then measured eta in fully hydrated bilayers composed of dioleoylphosphatidylcholine and dioleoylphosphatidylethanolamine (DOPC and DOPE respectively). Although the resolution of our measurements of lateral pressure as a function of distance into the monolayer was limited, we did observe a dip in the excimer signal in the region of the DOPC/DOPE cis double bond. As we isothermally increased the DOPE composition, and hence the desire for interfacial curvature, we observed, as expected, that the net excimer signal increased. However this net increase was apparently brought about by a transfer of pressure from the region around the glycerol backbone to the region near the chain ends, with the lateral pressure dropping above the cis double bond but increasing at a greater rate beyond the double bond. PMID- 10822600 TI - Interfacial enzyme activation, non-lamellar phase formation and membrane fusion. Is there a conducting thread? AB - Previous studies from this laboratory have shown that the enzymic generation of diacylglycerol in bilayers by phospholipase C may lead to membrane fusion through the formation of transient non-lamellar lipidic intermediates. The present paper intends to explore the correlations existing among the three main processes involved, namely (a) the induction (or inhibition) of lamellar-to-non-lamellar phase transitions in lipid mixtures through the addition of small (< 5 mol%) proportions of other lipids, (b) the promotion, by the latter lipids, of fusion in otherwise stable phospholipid vesicles (large unilamellar liposomes) under conditions leading to inverted hexagonal/inverted cubic phase formation in bulk lipid systems, and (c) the modulation, by the same small proportions of lipids, of phospholipase C hydrolysis of phosphatidylcholine in liposome bilayers. It is concluded that phospholipase C may give rise to non-lamellar lipidic structures that in turn permit liposomal fusion to occur, but neither enzyme activity is directly modulated by non-lamellar phase formation, nor will whatever kind of enzyme-induced non-lamellar structure give rise to fusion. Moreover, only under certain kinetic conditions will the enzyme give rise to the organization of non lamellar structures that are conducive to the fusion event. PMID- 10822601 TI - Characterization of the physical properties of model biomembranes at the nanometer scale with the atomic force microscope. AB - Interaction forces and topography of mixed phospholipid-glycolipid bilayers were investigated by atomic force microscopy (AFM) in aqueous conditions with probes functionalized with self-assembled monolayers terminating in hydroxy groups. Short-range repulsive forces were measured between the hydroxy-terminated probe and the surface of the two-dimensional (2-D) solid-like domains of distearoyl phosphatidylethanolamine (DSPE) and digalactosyldiglyceride (DGDG). The form and range of the short-range repulsive force indicated that repulsive hydration/steric forces dominate the interaction at separation distances of 0.3 1.0 nm after which the probe makes mechanical contact with the bilayers. At loads < 5 nN the bilayer was elastically deformed by the probe, while at higher loads plastic deformation of the bilayer was observed. Surprisingly, a short-range repulsive force was not observed at the surface of the 2-D liquid-like dioleoylphosphatidylethanolamine (DOPE) film, despite the identical head groups of DOPE and DSPE. This provides direct evidence for the influence of the structure and mechanical properties of lipid bilayers on their interaction forces, an effect which may be a major importance in the control of biological processes such as cell adhesion and membrane fusion. The step height measured between lipid domains in the AFM topographic images was larger than could be accounted for by the thickness and mechanical properties of the molecules. A direct correlation was observed between the repulsive force range over the lipid domains and the topographic contrast, which provides direct insight into the fundamental mechanisms of AFM imaging in aqueous solutions. This study demonstrates that chemically modified AFM probes can be used in combination with patterned lipid bilayers as a novel and powerful approach to characterize the nanometer scale chemical and physical properties of heterogeneous biosurfaces such as cell membranes. PMID- 10822602 TI - Modelling and simulation of light-activated membrane proteins: dynamical transitions in bacteriorhodopsin. AB - Many of the functions of membranes are carried out by proteins associated with them. A knowledge of atomic-detail membrane protein structures and dynamics is required for a full understanding of these functions. We briefly discuss recent progress in this field using modelling and simulation. One of the best characterised membrane proteins, bacteriorhodopsin, undergoes dynamical transitions with temperature. Here we present preliminary results of molecular dynamics simulation of this protein as a function of temperature, indicating the presence of dynamical transitions at approximately the temperatures seen experimentally. PMID- 10822603 TI - Interactions between poly(2-ethylacrylic acid) and lipid bilayer membranes: effects of cholesterol and grafted poly(ethylene glycol). AB - The exchange of the protonatable polymer, poly(2-ethylacrylic acid) (PEAA), has been studied with vesicle membranes containing cholesterol from 0 to 60 mol% or PEG2000-lipid (5 mol%). The release of an entrapped dye from 100 nm extruded liposomes was used as an assay for membrane perturbation by the polymer as a function of pH. The inclusion of cholesterol was found to reduce the pH at which the polymer caused release of the dye from the lipid vesicles, and the degree of polymer protonation (i.e., degree of hydrophobicity) correlated well with the increase in elastic expansion modulus of the vesicle bilayer. The results are discussed in terms of a balance between polymer solubility and membrane expansion. With respect to the PEG barrier, the presence of 5 mol% PEG2000, which represents full surface coverage, did not prevent PEAA from inducing contents release, demonstrating that highly hydrated polymeric layers are not effective barriers for other water soluble polymers, and may point to some association between the two polymers. PMID- 10822604 TI - Membrane electroporation and electromechanical deformation of vesicles and cells. AB - Analysis of the reduced turbidity (delta T-/T0) and absorbance (delta A-/A0) relaxations of unilamellar lipid vesicles, doped with the diphenylhexatrienyl phosphatidylcholine (beta-DPH pPC) lipids in high-voltage rectangular electrical field pulses, demonstrates that the major part of the turbidity and absorbance dichroism is caused by vesicle elongation under electric Maxwell stress. The kinetics of this electrochemomechanical shape deformation (time constants 0.1 < or = tau/microsecond < or = 3) is determined both by the entrance of water and ions into the bulk membrane phase to form local electropores, and by the faster processes of membrane stretching and smoothing of thermal undulations. Moreover, the absorbance dichroism indicates local displacements of the chromophore relative to the membrane normal in the field. The slightly slower relaxations of the chemical turbidity (delta T+/T0) and absorbance (delta A+/A0) modes are both associated with the entrance of solvent into the interface membrane/medium, caused by the alignment of the bipolar lipid head groups in one of the leaflets at the pole caps of the vesicle bilayer. In addition, (delta T+/T0) indicates changes in vesicle shape and volume. The results for lipid vesicles provide guidelines for the analysis of electroporative deformations of biological cells. PMID- 10822605 TI - Lipid-protein interactions in the membrane: studies with model peptides. AB - We have used fluorescence quenching of tryptophan-containing trans-membrane peptides by bromine-containing phospholipids to study the specificity of peptide lipid interactions. We have synthesized peptides Ac-K2GLm WLnK2A-amide where m = 7 and n = 9 (L16) and m = 10 and n = 12 (L22). Binding constants of L22 for dioleoylphosphatidylserine [di(C18 : 1)PS] or dioleoylphosphatidic acid [di(C18 : 1)PA] relative to dieoleoylphosphatidylcholine [di(C18 : 1)PC] were close to 1. However, for L16, whilst the bulk of the di(C18 : 1)PA molecules bound with a binding constant relative to di(C18 : 1)PC close to 1, a small number of di(C18 : 1)PA molecules bound much more strongly. Assuming just one high affinity binding site on L16 for anionic lipid, the affinity of the site for di(C18 : 1)PS was calculated to be ca. 8 times that for di (C18 : 1)PC. The relative binding constant was little affected by ionic strength and close contact between the anionic headgroup of di(C18 : 1)PS and a lysine residue on the peptide was suggested. The relative binding constant for di(C18 : 1)PS at this high affinity site was less than for di(C18 : 1)PA. Cholesterol interacts with L22 with an affinity about 0.7 of that of di(C18 : 1)PC. The structure of the peptide itself is important. The peptide Ac-KKGYL6WL8YKKA-amide (Y2L14) incorporated into bilayers of dinervonylphosphatidylcholine [di(C24 : 1)PC] whereas L16 did not incorporate into this lipid. It is suggested that thinning of a lipid bilayer around a peptide to give optimal hydrophobic matching is less energetically unfavourable when a Tyr residue is located in the lipid/water interfacial region. PMID- 10822606 TI - Phospholipid chain length alters the equilibrium between pore and channel forms of gramicidin. AB - Gramicidin is an excellent model system for studying the passage of ions through biological membranes. The conformation of gramicidin is well defined in many different solvent and lipid systems, as are its conductance and spectroscopic properties. It is a polymorphic molecule that can adopt two different types of structure, the double helical "pore" and the helical dimer "channel". This study investigated the influence of the acyl chain length of membrane phospholipids on the conformations adopted by gramicidin. We used circular dichroism spectroscopy to examine the conformational equilibrium between the pore and channel forms in small unilamellar vesicles of phosphatidylcholine with acyl chain lengths of 18, 20 and 22 carbons. Our results show that in C18 and C20 lipids almost all the gramicidin is in the channel form, while in the longer C22 lipids the equilibrium shifts in favour of pore conformations, such that they form up to 43% of the total population. This change is attributed to the ability of the double helical conformation to tolerate more hydrophobic mismatch than the helical dimer, perhaps due to the greater number of stabilising intermolecular hydrogen bonds. PMID- 10822607 TI - Protein inclusion in lipid membranes: a theory based on the hypernetted chain integral equation. AB - A theory for describing the structure of the hydrocarbon chains around a protein inclusion embedded in a lipid bilayer is developed on the basis of the hypernetted chain integral equation formalism for liquids. The exact lateral density-density response function of the hydrocarbon core, which is extracted from a molecular dynamics simulation of a pure lipid bilayer, is used as input to the theory. Numerical calculations show that the average lipid order is perturbed over a distance of 25 to 30 A around a hard repulsive cylinder of 5 A radius representing an alpha-helical polyleucine protein inclusion. The lipid-mediated protein-protein interaction is calculated and is shown to be non-monotonic, being repulsive at an intermediate range but attractive at short range. It is found that the lipid matrix contributes a free energy well of 8 kBT to the association of two cylindrical inclusions. PMID- 10822608 TI - Lipid packing stress and polypeptide aggregation: alamethicin channel probed by proton titration of lipid charge. AB - Lipid membranes are not passive, neutral scaffolds to hold membrane proteins. In order to examine the influence of lipid packing energetics on ion channel expression, we study the relative probabilities of alamethicin channel formation in dioleoylphosphatidylserine (DOPS) bilayers as a function of pH. The rationale for this strategy is our earlier finding that the higher-conductance states, corresponding to larger polypeptide aggregates, are more likely to occur in the presence of lipids prone to hexagonal HII-phase formation (specifically DOPE), than in the presence of lamellar L alpha-forming lipids (DOPC). In low ionic strength NaCl solutions at neutral pH, the open channel in DOPS membranes spends most of its time in states of lower conductance and resembles alamethicin channels in DOPC; at lower pH, where the lipid polar groups are neutralized, the channel probability distribution resembles that in DOPE. X-Ray diffraction studies on DOPS show a progressive decrease in the intrinsic curvature of the constituent monolayers as well as a decreased probability of HII-phase formation when the charged lipid fraction is increased. We explore how proton titration of DOPS affects lipid packing energetics, and how these energetics couple titration to channel formation. PMID- 10822609 TI - Structure-based prediction of the conductance properties of ion channels. AB - The HOLE procedure allows the prediction of the absolute conductance of an ion channel model from its structure. The original prediction method uses an empirically corrected Ohmic method. It is most successful, with predictions being reliable to within a factor of two. A new modification of the procedure is presented in which the self-diffusion coefficients of water molecules from molecular dynamics simulation are used to replace the empirical correction factor. A "prediction" of the conductance for the porin OmpF by the new method is made and shown to be very close to the experimental value. HOLE also allows the prediction of the effect that the addition of non-electrolyte polymers will have on channel conductance. The method has great potential to yield structural information from data provided by single channel recordings but needs further validation by making measurements on channels of known structure. Preliminary results are given of single channel records establishing the effects of non electrolytes on the conductance of gramicidin D channels. As an example of the potential uses of the procedure application is made to examine the oligomerization of alpha-toxin (alpha-hemolysin) channels. A model for the alpha toxin hexamer, based on the crystal structure for the heptamer, is generated using molecular mechanics methods. The compatibility of the structures with single channel conductance data is assessed using HOLE. PMID- 10822610 TI - Molecular dynamics simulation of a hydrated diphytanol phosphatidylcholine lipid bilayer containing an alpha-helical bundle of four transmembrane domains of the influenza A virus M2 protein. AB - An alpha-helical bundle composed of four transmembrane portions of the M2 protein from the Influenza A virus has been studied in a hydrated diphytanol phosphatidylcholine bilayer using molecular dynamics (MD) calculations. Experimentally, the sequence utilized is known to aggregate as a four-helix bundle and act as a pH-gated proton-selective ion channel, which is blocked by the drug amantadine hydrochloride. In the presented simulation, the ion channel was initially set up as a parallel four-helix bundle. The all-atom simulation consisted of almost 16,000 atoms, described classically, using a forcefield from the CHARMM22 database. Bilayers with and without the bundle were shown to be stable throughout the nanosecond timescale of the MD simulation. Structural and dynamical properties of the bilayer both with and without the transmembrane protein are reported. PMID- 10822611 TI - Alamethicin channels in a membrane: molecular dynamics simulations. AB - Alamethicin (Alm) is a 20 residue peptide which forms a kinked alpha-helix in membrane and membrane-mimetic environments. Ion channels formed by intramembraneous aggregates of Alm are thought to be formed by bundles of approximately parallel Alm helices surrounding a central bilayer pore. Different channel conductance levels correspond to different numbers of helices per bundle, ranging from N = 5 to N > 8. Calculation of the predicted pKA values of the ring of Glu18 sidechains at the C-terminal mouth of the pore suggests that at neutral pH most or all of these sidechains will remain protonated. Nanosecond molecular dynamics (MD) simulations of N = 5, 6, 7 and 8 bundles of Alm helices in a POPC bilayer have been run, corresponding to a total simulation time of 4 ns. These simulations explore the stability and conformational dynamics of these helix bundle channels when embedded in a full phospholipid bilayer in an aqueous environment. The structural and dynamic properties of water in these model channels are examined. As in earlier in vacuo simulations (J. Breed, R. Sankararamakrishnan, I. D. Kerr and M. S. P. Sansom, Biophys. J., 1996, 70, 1643) the dipole moments of water molecules within the pores are aligned antiparallel to the helix dipoles. This helps to contribute to the stability of the helix bundles. PMID- 10822612 TI - Kinetics of the competitive response of receptors immobilised to ion-channels which have been incorporated into a tethered bilayer. AB - A competitive ion channel switch (ICS) biosensor has been modelled yielding ligand mediated monomer-dimer reaction kinetics of gramicidin (gA) ion-channels within a tethered bilayer lipid membrane. Through employing gramicidin A, functionalized with the water-soluble hapten digoxigenin, it is possible to cross link gramicidin to antibody fragments tethered at the membrane/aqueous interface. The change in ionic conductivity of the channel dimers may then be used to measure the binding kinetics of hapten-protein interactions at the membrane surface. The approach involves measuring the time dependence of the increase in impedance following the addition of a biotinylated antibody fragment (b-Fab'), which cross-links the functionalized gramicidin monomers in the outer layer of the lipid bilayer to tethered membrane spanning lipid. The subsequent addition of the small molecule digoxin, (M(r) 781 Da), competes with and reverses this interaction. The model provides a quantitative description of the response to both the cross-linking following the addition of the b-Fab' and the competitive displacement of the hapten by a water-soluble small analyte. Good agreement is obtained with independent measures of the cross-linking reaction rates of the gramicidin monomer-dimer and the b-Fab: hapten complex. The rate and amplitude of the competitive response is dependent on concentration and provides a fast and sensitive detection technique. Estimates are made of the concentration of gramicidin monomers in both the inner and outer monolayer leaflets of the membrane. This is used in the calculation of the gramicidin monomer/dimer equilibrium constant, K2D3. Other considerations include the membrane impedance limit set by the membrane leakage which is also a function of the concentration of the gA monomer concentration, and the two-dimensional kinetic association constant k2D2, of the hapten: b-Fab' complex. The gA dimer concentration is dependent on both the concentration of gA-dig and of the tethered streptavidin: b Fab' complexes. The model shows that the 2D dissociation constant k2D3(-1), must be at least 10 times faster than the 3D dissociation constant k3D2(-1) for digoxin to completely reverse the cross-linked hapten-receptor interaction at the membrane interface. PMID- 10822613 TI - Three-dimensional models of glutamate receptors. AB - Structural models of glutamate receptors have been produced as part of a multidisciplinary study of neuronal function--both ligand/receptor interactions and ion transport--at the atomic level. The models have concentrated on the agonist binding and transmembrane domains of ionotropic (i.e. ligand-gated) glutamate receptors (iGluRs), and have aided our understanding of the molecular determinants of (1) ligand binding and (2) channel activity. The model building process involved a combination of homology modelling, distance geometry, molecular mechanics, protein-ligand and protein-protein docking, electrostatic calculations and manual adjustment, in conjunction with restraints from site directed mutagenesis, ligand binding and electrophysiological studies. The initial models were used to produce hypotheses which were tested experimentally; these models have been subsequently refined as part of an extremely effective multidisciplinary study using an iterative molecular modelling/experimental verification cycle in which restraints derived from experimental studies are used at all stages, and the findings from one round of modelling are used as restraints in the next. By studying a variety of agonists and antagonists, details have been built up of (1) those residues involved in ligand binding and (2) the role of agonist binding (i.e. agonist-induced conformational change) in channel gating. The models also aid our understanding of the conductance properties of the channels. PMID- 10822614 TI - Functional immobilization of biomembrane fragments on planar waveguides for the investigation of side-directed ligand binding by surface-confined fluorescence. AB - A method for the functional immobilization of Na,K-ATPase-rich membrane fragments on planar metal oxide waveguides has been developed. A novel optical technique based on the highly sensitive detection of surface-confined fluorescence in the evanescent field of the waveguide allowed us to investigate the interactions of the immobilized protein with cations and ligands. For specific binding studies, a FITC-Na,K-ATPase was used, which had been labelled covalently within the ATP binding domain of the protein. Fluorophore labels of the surface-bound enzyme can be selectively excited in the evanescent field. A preserved functional activity of the immobilized enzyme was only found when a phospholipid monolayer was preassembled onto the hydrophobic chip surface to form a gentle, biocompatible interface. In situ atomic force microscopy (AFM) was used to examine and optimize the conditions for the lipid and membrane fragment assembly and the quality of the formed layers. The enzyme's functional activity was tested by selective K+ cation binding, interaction with anti-fluorescein antibody 4-4-20, phosphorylation of the protein and binding of inhibitory ligand ouabain. The comparison with corresponding fluorescence intensity changes found in bulk solution provides information about the side-directed surface binding of the Na,K ATPase membrane fragments. The affinity constants of K+ ions to the Na,K-ATPase was the same for the immobilized and the non-immobilized enzyme, providing evidence for the highly native environment on the surface. The method for the functional immobilization of membrane fragments on waveguide surfaces will be the basis for future applications in pharmaceutical research where advanced methods for exploring the molecular mechanisms of membrane receptor targets and drug screening are required. PMID- 10822615 TI - Analysis of membrane protein cluster densities and sizes in situ by image correlation spectroscopy. AB - Communication between cells invariably involves interactions of a signalling molecule with a receptor at the surface of the cell. Typically, the receptor is imbedded in the membrane and it is hypothesized that the binding of the signalling molecule causes a change in the state of aggregation of the receptor which, in turn, initiates a biochemical signal within the cell. Subsequently, many of the occupied receptors bind to membrane-associated structures, called coated pits, which invaginate and pinch off to form coated vesicles, thereby removing the receptors from the cell surface. The state of aggregation of membrane receptors is obviously in constant flux. Any useful approach to measuring the state of aggregation must, therefore, allow for dynamic measurements in living cells. It is possible to use fluorescently labelled signalling molecules or antibodies directed at the receptor of interest to visualize the receptor on the cell surface with a fluorescence microscope. By employing a laser confocal microscope, high resolution images can be produced in which the fluorescence intensity is quantitatively imaged as a function of position across the surface of the cell. Calculations of autocorrelation functions of these images provide direct and accurate measures of the density of fluorescent particles on the surface. Combined with the average intensity in the image, which reflects the total average number of molecules, it is possible to estimate the degree of aggregation of the receptor molecules. We refer to this analysis as image correlation spectroscopy (ICS). We show how ICS can be used to measure the density of several receptors on a variety of cells and how it can be used to measure the density of coated pits and the number of molecules per coated pit. We also show how the technique can be used to monitor fusion of virus particles to cell membranes. Further, we illustrate that, by calculating cross correlation functions between pairs of images, we can extend the analysis to measurements of the distributions as a function of time, on the second timescale, as well as to measurements of the movement of the receptor aggregates on the surface. Finally, we illustrate that, by this approach, we can measure the extent of interaction between two different receptors as a function of time. This represents the most quantitative measurement of the extent of co-localization of receptors available and is independent of the spatial resolution of the confocal microscope. The theory of ICS and image cross-correlation spectroscopy (ICCS), focussing on the interpretation of the data in terms of the biological phenomenon being probed, is discussed. PMID- 10822616 TI - Direct measurement of recognition forces between proteins and membrane receptors. AB - The interactions between the protein, cholera toxin B subunit attached to an atomic force microscope, AFM, cantilever, CTB and its receptor the ganglioside, GM1 have been measured in a dilute electrolyte solution, pH 5.5. Although there is variation in the force separation data obtained, particularly on approach of the AFM tip to the GM1 surface where usually, but not always an attraction is noted, an adhesion is always noted on separation of the surfaces. The strength of this adhesion varies from experiment to experiment, but appears to be quantised at a value of around 90 pN. Addition of cholera toxin to the aqueous electrolyte solution completely removes the attractive interaction and adhesion. This gives us confidence that in the earlier experiments, a specific interaction between the CTB and GM1 was measured. PMID- 10822617 TI - Use of a laminar flow chamber to study the rate of bond formation and dissociation between surface-bound adhesion molecules: effect of applied force and distance between surfaces. AB - It has recently been shown that much information on the behaviour of surface bound adhesion molecules could be obtained by monitoring the motion of receptor coated particles along ligand-derivatized surfaces in the presence of a hydrodynamic force of a few pN. This procedure is expected to allow direct monitoring of the formation and dissociation of individual bonds. We present experimental results on the interaction between streptavidin-coated spheres (1.4 microns diameter) and control or biotinylated mica surfaces in a laminar flow chamber. Moving spheres are found to display numerous arrests whose frequency is markedly increased (5-13-fold) in the presence of biotin groups. For a given shear rate, the binding frequency is strongly dependent on the sphere-surface separation. Indeed, this frequency displayed a 14-fold decrease when the velocity increased from 7 to 15 microns s-1 for a wall shear rate of 20 s-1. Furthermore, the lifetime of observed arrests was of the order of several seconds, i.e. 5-50 fold higher than previously determined on models such as selectin-ligand, CD2 CD48 or cadherin-cadherin. Finally, this lifetime did not decrease when the wall shear rate was increased from ca. 10 to 40 s-1. PMID- 10822618 TI - Concluding remarks and the challenge from the immune system. AB - This meeting has covered a wide range of approaches to the study of interactions of biomembranes and one is struck by the progress in their analysis and the application of many new methods. The main theme of the meeting has been the structure of biomembranes and their components. Rather than pick highlights of these or attempt to summarise the findings in these Concluding remarks, I will instead summarise some of the interactions of the cells of the immune systems for which immunologists would like explanations at a molecular level. One of the features of the immune system is that it involves a variety of populations of cells that have complex migratory patterns and interactions that occur throughout life. The surfaces of these cells--the leukocytes--mediate interactions that are essential for the fine control of the immune system that ensures the rapid but controlled rejection of foreign materials such as viruses and bacteria. At the same time it must ensure that reactivity against self is prevented, otherwise autoimmune diseases such as rheumatoid arthritis and multiple sclerosis may result. Some of the features of the interactions involved are outlined in this short overview with more detailed analysis of the leukocyte cell surface given in ref. 1. PMID- 10822619 TI - [The characteristics of the functional state of the human brain anatomical formations during different levels of reactive anxiety. A PET study of healthy volunteers]. PMID- 10822620 TI - [The principles of complementarity and commensurability as the basis of new research technologies in the field of the neurophysiology of functional states]. PMID- 10822621 TI - [The reorganization of the spatial EEG correlation evoked by visual stimulation]. PMID- 10822622 TI - [Changes in the interhemispheric interaction in patients with an affective pathology and schizophrenia]. PMID- 10822623 TI - [The neurophysiological aspects of the compensatory-recovery processes during the cerebrospinal fluid therapy of central visual disorders]. PMID- 10822624 TI - [The discrimination of angular velocity with the sound source moving in a vertical plane]. PMID- 10822625 TI - [The age-sex traits of the perception of the emotional characteristics of speech under noise exposure]. PMID- 10822626 TI - [The inversion of hemispheric dominance as a psychophysiological mechanism in intervalic hypoxic training]. PMID- 10822627 TI - [Motivation as a factor in the development of an operator's functional state of tension]. PMID- 10822628 TI - [Autonomous control of the heart rate in patients with ischemic heart disease as related to concomitant pathology or complications]. PMID- 10822629 TI - [The types of adaptation of the cardiorespiratory functions in athletes to physical loading]. PMID- 10822630 TI - [The correlation of skeletal muscle force-stiffness depends on the level of activation of the contractile apparatus and on the prehistory of contraction]. PMID- 10822631 TI - [The effect of additional support on the vibration-induced reactions of human beings in a standing posture]. PMID- 10822632 TI - [Voluntary recruitment of the motor units during body overheating]. PMID- 10822633 TI - [The hormonal mechanisms supporting muscle work during prolonged head-down tile hypokinesia]. PMID- 10822634 TI - [The effect of normobaric hypoxia on the blood growth hormone level in adults]. PMID- 10822635 TI - [The characteristics of the sounds of normal human forced expiration]. PMID- 10822637 TI - [The nature of the metabolic changes in workers under a field-watch work regimen in the Polar regions]. PMID- 10822636 TI - [The lipid metabolic status of the population of children in the southern Maritime Territory]. PMID- 10822638 TI - National Committee on Assisted Human Reproduction: a promising first step. PMID- 10822639 TI - A model ethical protocol as a guidance document for human genome diversity research. PMID- 10822640 TI - [Legal and philosophical considerations about cloning in human reproduction]. AB - The birth of Dolly the sheep has served to rekindle the debate on the lawfulness of cloning for human reproduction. Although the normative response has been unanimous thus far in sanctioning such use, the matter of the limits to this is widely debated. In this article the author examines the reasons underpinning the limits imposed: is cloning for human reproduction unlawful in itself of does everything depend on the aim pursued? The issue is addressed from the standpoint of the apparent clash in law between the right to reproduce and freedom of research, on the one hand, and--on the other--the right to be conceived in a given manner. Particular attention is paid to the question of the lawfulness of two instances of human embryo division, namely, for in vitro fertilization and for preimplantation diagnosis. PMID- 10822641 TI - [Genetic research as evidence in the German criminal process after the Criminal Law Statute of March 17, 1997 (Part II)]. PMID- 10822643 TI - Human freedom and the human genome (Part I). PMID- 10822642 TI - [Legal limits to the alteration of the human genetic heritage (Part II)]. PMID- 10822644 TI - [The second decision pronounced by the First Tribunal of the Supreme Court in a case of wrongful birth (February 4, 1999). Does it contradict the resolution of the decision pronounced on June 6, 1997 about the same problem?]. AB - This paper serves a twin aim: firstly, to examine in detail and give an opinion on the ruling of the Supreme Court of 4 February 1999, and secondly, given the contradiction between this ruling and a previous one handed down by the same court (6 June 1997), to set out a number of considerations on the problem addressed in both cases. PMID- 10822645 TI - [Comment on the Decision of the Constitutional Court 212/1996 of December 19, 1996 (Part II)]. PMID- 10822646 TI - [Social perception of biotechnology: a new tool for decision making]. AB - The increase in critical opinions of scientific and technological development in advanced societies has led to the development of studies examining the links between science, technology and society. Among these studies are those on public perception, a particularly topical issue being the perception of biotechnology. There has been considerable European activity in this area of reflection and research, as evidenced by the wide range of initiatives at European Union and member state level. Some special instruments, such as the consensus conferences which began in Scandinavian countries, are now being applied in various EU countries. In Spain several public bodies and foundations are involved in a growing number of activities in this field. Biotechnology has become a particularly interesting research area for promoting new conditions for analysis of the social and economic repercussions of advances in science and technology. The study of the social perception of biotechnology is a crucial instrument for policy design and management and hence a methodological review is warranted. PMID- 10822647 TI - Biopolitics--the bio-environment. Biocentric ethics in the next millennium. PMID- 10822648 TI - Ethical aspects of research involving the use of human embryo in the context of the Fifth Framework Programme. PMID- 10822649 TI - [First report of the National Commission on Assisted Reproduction: general summary]. PMID- 10822650 TI - Spain's Constitutional Court and assisted reproduction techniques. PMID- 10822651 TI - [Genetic information and knowledge]. AB - This presentation has a double aim of clarification: epistemological and ethical. The analysis mainly refers to the uses and representations of genetic knowledge regarding people or groups. The descriptive part is completed with ethical stands regarding the following issues: transindividual scope of genetic knowledge, the obligation to know and the right not to know, discrimination and solidarity, and appropriation of genetic knowledge. PMID- 10822652 TI - [Right to privacy, reservation or secret. Changes in perspective of research on human genome]. AB - 1. Three kinds of privacy can be identified: territorial, bodily and, thirdly, psychological or spiritual. Cases and examples of each. 2. Two aspects are involved in breaches of the right to privacy--in any of its three forms-: the actual invasion of someone's privacy and the subsequent publication of the product thereof. 3. The right to privacy can enter into conflict with other constitutionally protected right such as the right to information, freedom of expression and freedom of scientific enquiry. 4. Research on the human genome has opened up a new area in privacy, given that it is unlawful to intrude into the genetic structure of a person without said person's lawful consent. Nothing is more private than a person's own individual genetic code. There may be exceptions to the right to genetic privacy: cases of lawful intrusion. 5. Unwarranted intrusion: discrimination in school admissions, employment contracts and insurance policies. PMID- 10822653 TI - Human freedom and the human genome (Part II). PMID- 10822654 TI - Genetic counseling and the best interest of the child. PMID- 10822655 TI - [Decision of the Constitutional Court 116/1999 of June 17, 1999. Appeal on unconstitutionality against Law 35/88, of November 22, regarding Assisted Reproduction Techniques]. PMID- 10822656 TI - [The Constitutional Court regarding Assisted Reproduction Techniques: a critical assessment]. AB - Last June Spain's Constitutional Court ruled on the challenge lodged against Law 35/1988 on Assisted Reproduction Techniques (Ruling 116/1999). The ruling confirmed the provisions of the Law. However, in examining the Law in question the Court did not, in my opinion interpret correctly the Spanish Constitution. I argue that there are three main areas of disagreement: since the contents directly affect human dignity the Law should have taken the form of a higher (Organic) Law. Secondly, the Law's treatment of in vitro fertilised embryos in certain cases, falls short of the minimum required by the Constitution. Thirdly, paternity investigation is prevented contrary to the provisions of the Constitution and hence the constitutional notion of family is undermined. PMID- 10822657 TI - [Genetic reading of the decision of the Constitutional Court on the appeal on unconstitutionality against Law 35/1988 on Assisted Reproduction Techniques]. AB - In this article we provide a "genetic reading" of the challenge on grounds of alleged inconstitutionality made against Spain's Law on Assisted Reproduction Techniques (Law 38/1988) and of the ruling handed down by the Constitutional Court on 17 June 1999. A critical appraisal is given of some genetic and biological concepts which, in the author's view, are used incorrectly in the legal texts presented. The article also shows that at the heart of the matter lies still the latent problem of the status in law of embryos. Lastly, brief reference is made to the issue of freedom of research. PMID- 10822659 TI - Managing genetic testing information: legal, ethical, and social challenges. PMID- 10822658 TI - [Genes and discrimination]. AB - The Human Genome Project (HGP) is the greatest scientific adventure in modern human biology, and the genetic map that is going to be revealed through this Project is going to be an important basis of the medicine of the future. Human beings do not however depend solely on their genes. In order to comprehend human pathology, it is essential to focus on the genetic factors and on the environmental factors. Genetic diagnoses, being fostered by the HGP, make it possible to know genetic predisposition and the risks of the onset of a given disorder. Predictive medicine offers great hopes, but is giving rise to major concerns and is causing ethics-related dilemmas. Confidentiality, the moral imperative of medicine, is necessary to prevent discriminatory deviations. As is stated in the UNESCO's Universal Declaration on the Human Genome and Human Rights, no one shall be subjected to discrimination based on genetic characteristics. PMID- 10822660 TI - [Genetic engineering and assisted reproduction techniques in man: a framework for sociologic analysis]. AB - The possibilities opened up by genetic engineering and assisted reproduction techniques require reflection by sociologists and extensive public debate. In view of their potential as factors of social change, evaluation and control are warranted. They can be viable only if transparent and through public co responsibility, for which an exchange of views is needed between all those who play a part in the development of said techniques. This dialogue must be wholly interdisciplinary and democratic. PMID- 10822661 TI - [Brazilian criminal law and genetics]. AB - This article analyses the Brasilia criminal regulation on genetic. Act No. 8.974/95 is examined because it regulates some criminal typologies on genetic engineering and assisted reproduction. Moreover, it presents information about the Act Project No. 149/97, on genetic discrimination. PMID- 10822662 TI - [The human genome and the insurance contract]. AB - The author studies the juridical implications that nowadays are appearing in relation to the human genome and health insurances. As we know it is really necessary to take steps to avoid some discrimination that would appear when it is known that a person suffer from an special disease. PMID- 10822663 TI - [The viruses that cause gastroenteritis]. PMID- 10822664 TI - [Combined preparations in the treatment of hypertension]. PMID- 10822665 TI - [Angiotensins and bradykinin in the vital processes of cells]. PMID- 10822666 TI - [The outlook for the use of EHF-range electromagnetic radiation in the treatment of background and precancerous diseases of the cervix uteri]. PMID- 10822667 TI - [Biliary tract diseases in persons suffering as a result of the accident at the Chernobyl Atomic Electric Power Station]. AB - A retrospective analysis was performed of case histories and of results of sonographic investigations in liquidators of the Chernobyl accident suffering from chronic abnormalities of the biliary ducts. Patients with cholecystitis were studied for the biochemical composition of their bile. The incidence of the gallbladder disorders (chronic cholecystitis, angiocholitis, dyskinesias of the biliary ducts) has not changed much over the last 10 years having elapsed since the accident. The biochemical composition of bile was found to have been changed to a greater extent in the liquidators of the Chernobyl accident than it was in those having avoided danger of exposure to ionizing radiation. Mechanisms of origination of cholelithiasis are discussed on the basis of investigations designed to study biochemical properties of bile and findings secured with the aid of the ultrasound techniques. PMID- 10822668 TI - [Nonrespiratory lung functions in children who have undergone the chronic effect of ionizing radiation as a result of the accident at the Chernobyl Atomic Electric Power Station]. AB - With the purpose of assessing the condition of nonrespiratory functions of the lungs in those children having been under chronic exposure to ionizing irradiation as a result of the Chernobyl accident in the time-related course of the post-accident period, an examination was done of 964 children free from clinically manifest pathology of the respiratory organs aged 5 to 14 years, residing in the zones of radiation control. Noted in the above children over the post-accident period are changes in indices for non-respiratory functions of the lungs suggesting diminution of respiratory moisture oozing, disordered production of pulmonary surfactant, with intensity of free-radical processes being rather high. PMID- 10822669 TI - [Health resort factors in the regulation of sexual disorders in men who permanently live in a radionuclide-contaminated area]. AB - The paper presents a clinical material on effects of health-resort factors, hyperbaric oxygenation, and psychotherapeutic correction on the gonadal function in patients having sexual problems. The multimodality approach permitted not only a significant alleviation of pathological symptomatology on the part of the autonomic nervous system, pain syndrome, improving the consistency and function of the prostate, but also had a beneficial effect on the copulative sphere, indices for microscopy of the prostatic secretum, spermatogenesis, and fertility of the ejaculate. PMID- 10822670 TI - [The anthropometric indices of newborn infants in the Kryvyy Rih industrial region]. AB - A study was made of anthropometric measures (body mass, length of the body, circumference of the head, circumference of the chest) in infants borne to women in separate primi-, secundi-, or multipara events in those areas of the Krivoy Rog industrial region arbitrarily taken as relatively pure (unpolluted) and those affected by radioactive contamination, with ages of the child-bearing women being 18-25, 26-30, and greater than 30 years old. Newborn babies from the contaminated district of the city were found to have higher anthropometric values. There was a persistent augmentation of the circumference of the chest in newborns from the contaminated district, which fact suggests development of adaptive processes in the fetus during the intrauterine development under conditions of contaminated atmosphere of the industrial region. PMID- 10822671 TI - [The hemostatic potential of the venous and arterial beds in patients with destabilized ischemic heart disease]. AB - Coagulating activity was studied of the arterial and venous blood platelet-rich and platelet-poor plasma in 16 patients with unstable angina (UA). An unusual activation of the hemostatic potential was recordable in the venous and arterial beds, as per changes in the general index of thrombophilia, which fact, along with the presence of FDR and fibrin-monomer complexes, can be regarded as signs of disseminated intravascular coagulation which can cause thrombogenic complications in UA patients. Changes in the system of hemostasis in the above patients (as evidenced by the index of thrombophilia) might suggest the prethrombotic state. The results obtained showed that platelets had an important role in the formation of the hemostatic potential both in the venous and arterial blood in UA. PMID- 10822672 TI - [The indices of intracardiac hemodynamics in patients with postinfarct cardiosclerosis]. AB - Endocardiac hemodynamics was studied in 81 patients with postinfarction cardiosclerosis with intact and decreased ejection fraction (EF > 57% and EF < 57% respectively) using echocardiographic B- and M-mode techniques. Patients with postinfarction cardiosclerosis without overt heart failure demonstrated a decline in the left ventricular pump function which was more manifest in postinfarction cardiosclerosis presenting with a decreased ejection fraction. A decline in the pump function of the myocardium in those patients having postinfarction cardiosclerosis without overt cardiac insufficiency is an indication for prescription of those medicinal substances enhancing the inotropic function of the myocardium (cardiac glycosides, peripheral vasodilators). PMID- 10822673 TI - [The etiological structure of pericarditis]. AB - Data are submitted from the published literature and the authors' observations on causation of pericarditis and transformation of its patterns over the last 24 years. The paper is based on the analysis of 325 patients having been operated on for constrictive pericarditis during the period 1974-1998. Over the last 12 years there has been an increase in the incidence of pericarditis of nonspecific etiology whereas that of pericarditis of rheumatic and tuberculous genesis has gotten substantially decreased. PMID- 10822674 TI - [The assessment of the degree of the colonization of the gastric mucosa by Helicobacter pylori and of the gastritis and duodenitis activity in duodenal peptic ulcer in different age groups]. AB - The article contains intriguing data on the difference in H. pylori extents of growth in the gastric antrum mucosa of patients at various ages with duodenal ulcer (DU). A comparative evaluation was done of extents of growth, average antibody titre, and activity of gastritis of the body of antral gastritis and duodenitis in different age groups in DU. The author has come to the conclusion that in the given population, H. pylori extents of growth and activity of antral gastritis and duodenitis in DU young subjects are not lower but even higher than in those at later ages. PMID- 10822675 TI - [The clinical aspects of acute liver failure in leptospirosis]. AB - Acute hepatic insufficiency is the main link in the pathological process in leptospirosis determining the degree of severity of the condition and level of bodily compensatory potentialities. Pathogenetically, hepatic insufficiency runs its course through the following three stages: compensated, sub-compensated, and decompensated characterized by different levels of functional (excretory-biliary, protein-synthetizing, hemostatic, detoxicating) inadequacy. PMID- 10822676 TI - [The effect of immunocorrective preparations on the cellular immunity indices of patients with recurrent respiratory diseases]. AB - Sensitivity of T-lymphocytes to different immuno-correcting drug preparations was studied in 183 individuals. Of these, 82 were always ailing children who ranged from 2 to 15 years old. Differences were disclosed in the individual sensitivity of in vitro T-lymphocytes, related to age and condition of the immunoregulatory index. The secured results should be taken account of in prescription of immuno correcting preparations. PMID- 10822677 TI - [A comparative analysis of the neurophysiological mechanisms of hearing disorders of different origins by the indices of the background and evoked bioelectrical activity of the brain]. AB - The hearing function was studied as was the bioelectrical activity of the brain in workers exposed to noise in their occupations and in those persons coming into contact with ionizing radiation with normal and disordered hearing. The studies made showed that those persons under exposure to ionizing radiation exhibited more pronounced disturbances in the brain stem structures of the acoustic analyzer than those exposed to noise even if they were early in the course of the ear disorder. We consider it expedient to undertake studies on bioelectrical activity of the brain to gain further insights into the mechanisms of sensorineural hearing loss. PMID- 10822679 TI - [The societal danger from epilepsy patients with nonconvulsive paroxysmal states]. AB - 237 patients with epilepsy having committed 248 socially dangerous acts (SDA) during the period of nonconvulsive paroxysmal states (dysphorias, twilight sleep states of consciousness, schizophrenia-like psychoses) were studied by clinical psychopathological routines. During the above psychopathological periods epileptic patients committed their SDAs on relatively rare occasions. But a high specific weight of aggressive acts having been committed during these very periods including grave ones (against life, health, and dignity of the personality), the incidence of the above states encountered in the clinical practice of dealing with epilepsy patients together with a high probability of their committing SDAs during the periods concerned determine the above contingent of patients in periods of nonconvulsive paroxysmal states as being of high social danger. In all cases the character of SDA was determined from the psychopathological context of psychotic episodes. PMID- 10822678 TI - [The clinical monitoring of epileptic seizures]. AB - The Chalfont Scale of Cerebral Seizure Severity has been adapted for use in a computer case history and the making-decision support system in managing patients with cerebral seizures (CS). The index of severity of cerebral seizures (CSSI) has been worked out, it being taken as the sum of results of assessment of the same type CS according to the Chalfont Scale of Cerebral Seizure Severity multiplied by the frequency of the paroxysms for all types of fits occurring in the given patient. The frequency of the same type CS is an arbitrary value for the number of such fits occurring during the 24-hour period, it can be a fractional number. This value is calculated on the basis of a more prolonged period out of the following two time periods: time between the last two CS or time between the last seizure and the moment of the computations made. Using the above-described approach a physician or the particular information system can speedily assess an individual course of the malady and efficacy of the treatment option chosen. The graphic representation of SSI and of doses of antiepileptic drugs stated in terms of phenobarbital units helps in creating an integral idea of a particular case. The assessment by the information system of the time related clinical course of the disease in 30 CS patients with making use of CSSI was found out to be in full agreement with opinions of three experts. Based on the use of CSSI in a clinical monitoring of CS in 111 patients there has been determined a place for the above index among other characteristics of the illness with preliminary recommendations having been given on its interpretation and role in managing of CS patients. PMID- 10822680 TI - [The clinical picture, diagnosis and treatment of cytomegalovirus encephalitis in adults]. AB - Described for the first time in Ukraine are eleven cases of encephalitis caused by cytomegalovirus (CMV) in adults. Characteristic features of clinical presentation, laboratory and instrumental methods of investigation are presented with special emphasis being placed on predominant location of the pathological process in CMV encephalitis such as frequently encountered affection of the white substance of the brain predominantly in the paraventricular zones, significant intracranial liquor hypertension. Etiotropic treatment is to be prescribed in CMV encephalitis with gancyclovir (cymeven Roche) in the earliest periods of the course of the disease. PMID- 10822681 TI - [The clinical significance of morphological study of the skin in patients with diabetic microangiopathy of the lower extremities]. AB - Results are analyzed of a morphologic investigation designed to study the skin in 62 patients with diabetic microangiopathy of the lower extremities. Criteria are provided reflecting stage-dependent signs of the disorder by changes in the skin. In 23 patients, a possibility has also been explored of our using cryostat-aided sections in a rapid histological investigation (within 1 to 2 hours) to determine the stage of angiopathy. Morphological diagnosis is of particular value in patients with pyo-inflammatory processes in the inferior limbs in whom clinical verification is a challenge, in which case it proved to be helpful for the purpose of selecting a suitable treatment option as well. PMID- 10822682 TI - [The drug prevention of postoperative thromboses in patients with critical lower limb ischemia]. AB - Based on the author's findings a conclusion has been reached that it is expedient to use fraxiparin, a low-molecular heparin, in the early postoperative period after reconstructive operations on the arteries, and ticlid, a deaggregant on a regular basis, in the remote postoperative period, this drug having been found to increase the percentage of spared critically ischemized limbs compared with aspirin. PMID- 10822683 TI - [Patient medical rehabilitation following osteoplastic operations using hydroxylapatite-based porous ceramic]. AB - At the URIOP Department of Tumours of the Locomotor System, osseous plastic operations were performed with making use of hydroxilapatite-base porous ceramics in 60 patients with benign tumours of bone and tumour-like lesions of bone. Apart from surgery the patients underwent a complex of rehabilitative measures, such as remedial gymnastics, massage, designed to restore locomotor functions of the extremity. Time periods of medical rehabilitation of patients were found to have gotten reduced by 1 to 2 months on the average providing the multimodality treatments with remedial gymnastics and massage were administered. PMID- 10822684 TI - [The structural characteristics of the placental barrier during the dynamics of pregnancy in women over 35]. AB - Biopsy of chorion early in the course of pregnancy is of great diagnostic value. Morphological, electron microscopic, and morphometric findings for the shaggy chorion during the I to II trimesters of pregnancy show that in women aged 35 to 45 years with physiological pregnancy changes occur in all structures of the placental barrier as compared to young women. During the I and II trimesters of pregnancy, those women who range from 35 to 45 years old exhibit disturbances in the processes of chorion vascularization, with the compensatory and adaptive reactions developing on the cellular and subcellular levels. In women aged 41 to 45 years, disordered processes of vascularization in the placental barrier are recordable together with dystrophic changes in the syncytiotrophoblast and compensatory and adaptive reactions in endothelial cells of fetal capillaries. PMID- 10822685 TI - [The effect of combined therapy on the ortofen concentration of the blood plasma and synovial fluid in patients with rheumatoid arthritis]. AB - A pharmacologic investigation confirmed the possibility of deliberate correction of pharmacological properties of the nonsteroidal anti-inflammatory preparation orthophen in a clinical setting by combining it with those drugs (cimetidin and cocarboxylase) capable of inhibiting elimination of orthophen from the body. In consequence, the level of the drug gets elevated not only in blood plasma but also in the synovial fluid of those joints affected by inflammation, which fact secures high efficiency of the antiinflammatory therapy. PMID- 10822686 TI - [The ultrasonography of the capsular ligamentous apparatus of the knee joint in the early stages of rheumatoid arthritis]. AB - With the purpose of finding out informative value of the ultrasound investigation designed to study the capsular and ligamentous apparatus of the knee joint in its instability during the early stages of rheumatoid arthritis and correlating clinical symptoms with ultrasonographic findings an examination was done of twenty joints of patients in early stages of rheumatoid arthritis presenting with clinical signs of anterior-medial instability. Sonography confirmed the presence of instability and permitted the qualitative assessment of its degree to be done. The method allows us to disclose relative incompetence of the anterior-medial sector of the knee joint in those patients presenting with early stages of rheumatoid arthritis, which is one of causes of instability, with the cruciate and lateral ligaments remaining uninjured. Ultrasonography makes it possible to perform a quantitative assessment of the degree of instability of the joint irrespective of the clinical test used and experience of the orthopedist. PMID- 10822687 TI - [The characteristics of the course and treatment of patients with hypertension combined with gastric and duodenal ulcers and chronic gastroduodenitis]. AB - Overall, seventy-five patients with hypertensive disease concurrent with gastric and duodenal ulcer and chronic gastroduodenitis were examined. Hypertensive disease preceding peptic ulcer and chronic gastroduodenitis runs a graver course compared with that hypertensive disease having developed against the background of gastroduodenal pathologies, such as higher values for arterial pressure, peripheral resistance of arterioles, more pronounced changes in the eye-ground and heart. In treating patients with the above intercurrent illnesses a combination of the beta-adrenoblocker anaprilin and calcium antagonist phenihidin proved to be the most rational treatment option. PMID- 10822688 TI - [The pathogenetic and diagnostic characteristics of cholestatic forms of viral hepatitis]. AB - Some specificities of pathogenesis of cholestatic forms are submitted together with classification of causes of cholestatic syndrome in viral hepatitis. The problems of diagnosis of cholestatic forms of viral hepatitis are highlighted with special reference to specific features of the underlying pathological process, with special emphasis being placed on using those techniques sparing the injured parenchyma of the liver, biochemical and ultrasound ones among their number. Criteria have been established of ultrasound diagnosis for different forms of cholestasis in viral hepatitis. PMID- 10822689 TI - [The enteral syndrome in the clinical course of diffuse toxic goiter]. AB - Particular features have been studied of the clinical course of diffuse toxic goitre complicated by enteral syndrome. A total of 243 patients were examined. Of these, 139 were patients presenting with enteral disorders. Thyrotoxic enteropathy was diagnosed in 35 (52%) mild cases of thyrotoxicosis, the total number of such cases being 67, in 55 of 98 (56%) moderately severe, and 49 of 78 (63%) severe cases of thyrotoxicosis. PMID- 10822690 TI - [Lavage of the digestive tract with an isotonic mineral water-based 3-salt solution]. AB - The article presents technology, chief indications and contraindications, therapeutic effectiveness of peroral lavation of the alimentary canal with the drinking mineral water-base isotonic polyionic solution, its composition being as follows (g/dm3): sodium chloride, 6.5, potassium chloride, 0.75, sodium hydrocarbonate, 2.5. The patient takes on an empty stomach 5 to 7 l of the solution, t 40-37 degrees C, mean rate 50 ml/min. The procedure is done again in 7 to 10 days, the course of treatment involving 3 to 5 procedures, subsequently one procedure/30 to 60 days. After lavation the patient takes an enterosorbent and keeps to fruit and vegetable diet or has salt-, sugar-, and fat-free porridge. Kept under observation were 1384 patients 5 to 68 years old who had 3552 procedures done in them. The benefit from a complex sanatorium treatment involving the above procedure occurred in 26% of patients and 8% of controls. PMID- 10822691 TI - [Periodic gastric and duodenal motility in peptic ulcer patients after selective proximal vagotomy]. AB - The condition was studied of intermittent motility (IM) of the stomach and duodenal bulb in patients with duodenal ulcer in the preoperative period, six months later and 12 months after performing selective proximal vagotomy (n = 131, 103, and 83 respectively). All patients presented with disorders in their IM which were found to be dependent on their preoperative condition failing to reverse even 6 to 12 months after having the operation. PMID- 10822692 TI - [The use of prodigiozan in the combined therapy of the sequelae of mild craniocerebral trauma]. AB - Based on findings secured with the aid of instrumental and immunological techniques (electroencephalography, echoencephalography, roentgenoencephalography) a total of 57 patients with sequelae of mild craniocerebral injury were examined. Used in the study as an immunomodulating agent was prodigiosan. The drug was found to improve the bioelectrical activity of the brain, and to increase the amplitude of blood filling as evidenced by neurophysiological investigations. Moreover, prodigiosan makes for reduction of vegetovascular disorders and epileptic seizures, returning the level of neurospecific proteins to normal. PMID- 10822693 TI - [The diagnostic significance of immunogenetic markers in vertebrogenic neurological syndromes]. AB - A total of 116 patients presenting with different neurological syndromes of vertebral osteochondrosis were examined. More common in patients with reflex neurological manifestations of osteochondrosis compared with controls (n = 51) was a complete HLA phenotype (heterozygosity) determining better evolutional and biological resistance to injuring factors and ability to produce adequate adaptive and compensatory reactions. The diagnosing of incomplete phenotype in other groups indicated homozygosity of genes and suggested a high risk for development of the disorder. Criteria have been established for a relative risk of development of the disease for all six vertebrogenic neurological syndromes. PMID- 10822694 TI - [Allergic manifestations in diphtheria]. AB - Described in the article are particular characteristics of allergoses encountered in patients with different forms of diphtheria. Allergic reactions were found to be presenting as anaphylactic shock urticaria and Quincke's edema type. The studies made showed that allergic reactions in treatment of patients with diphtherial infection develop on relatively rare occasions but they should be examined in a more thorough way. PMID- 10822695 TI - [The content of hypophyseal-thyroid system hormones of the blood in pregnant women with late gestosis]. AB - A study made of thyroid hormone levels in healthy pregnant women and those presenting with late gestoses showed a significant increase in the latter of thyrotropine with simultaneous decrease being recordable in the levels of thyroxin and triiodthyronine. A correlation was established between the concentration of hormones of the thyroid gland and degree of severity of late gestosis. PMID- 10822696 TI - [Correction of the imbalance in the antioxidant protection system of the body in patients with severe gestosis]. AB - The role has been studied of thiol compounds as indicators of bodily antioxidant defence in patients with severe gestoses. A comprehensive evaluation was done of disorders in the system lipid peroxidation/antioxidant defence, with effectiveness of their correction with ozonohemotherapy and antioxidants included into the conventional intensive therapy of patients having been studied as well. The above therapy makes for restoration of intracellular environment of erythrocytes at the expense of maintenance and augmentation of the compensatory reaction in respect of activation of the enzymic link of the antioxidant defense in blood plasma of patients. PMID- 10822697 TI - [Antioxidants combined with enterosorption in the symptomatic treatment of ovarian cancer]. AB - As many as 36 patients with III-IV-stage ovarian carcinoma exposed to a symptomatic treatment were examined. The analysis of the studies made showed that the use of antioxidants and enterosorption combined in treatment of the above patients results in lessening of endogenous intoxication, provides immunoprotection, and improves the process of lipid peroxidation. PMID- 10822698 TI - [Active radiation telethermometry in the complex diagnosis of ovarian tumors]. AB - An active remote radiation thermometry was used in the diagnosis and differential diagnosis of ovarian tumours by determining the heat flow from the area of projection of the ovaries and control background. Overall fourty three patients with ovarian tumours were examined by this method. Significance of the results secured was verified during the histological technique-aided operation. The authors have come to the conclusion that remote thermometry involving loading tests is a simple supplementary method of investigation that is helpful in diagnosing of both benign and malignant ovarian tumours. PMID- 10822699 TI - [The effect of the preparation Wobenzym on the antioxidant protection indices and on the functional-morphological properties of the erythrocytes in a toxic lesion of the liver]. AB - Based on the analysis of results of examination of 60 patients with toxic affections of the liver particular features of the anti-radical defence decompensation mechanisms have been established together with changes in morphofunctional properties of red cells in toxic hepatitis in patients with pulmonary tuberculosis. It was considered expedient to include the polyenzymic preparation Wobenzyme into the therapeutic complex since it had been shown to make for normalization of the red cells' ability to undergo deformation, lowering the degree of viscosity of RBC suspension in the above patients. PMID- 10822700 TI - [The effect of symptomatic therapy with the alpha 1-adrenoblocker Cardura in patients with benign prostatic hyperplasia on the course of cardiovascular diseases]. AB - Forty-two patients with benign hyperplasia of the prostate were studied for the role of symptomatic therapy with a selective adrenoblocker Cardura in the prophylaxis of progression of such diseases of the cardiovascular system as arterial hypertension, angina pectoris. The obstructive and irritative symptomatology-related uresis has been shown to get normalized. Also recordable in the conducted investigations was an alleviation of arterial hypertension together with a noticeable decrease in the frequency and severity of attacks of angina. PMID- 10822701 TI - [The use of Ukrainian laferon in the combined treatment of patients with chronic myeloid leukemia]. AB - A study was made into the effect of laferon on effectiveness of treatment of patients with chronic myeloid leukemia (CML). The authors have reached the conclusion that drug tolerance in the patients is fairly good. The number of negative reactions within a few days of the start of laferon therapy did not exceed the level reported from other countries. Small series and short-term follow-up does not allow judgement about effects of the drug on survival but one can say with certainty that the employment of laferon in the treatment programme for CML in different categories of risk permits achieving clinical and hematological compensation on more frequent occasions. The quality of the patients' life gets improved. Chemotherapy was found to lessen the risk for infectious and inflammatory complications. PMID- 10822702 TI - [Disorders of central hemodynamics and water-salt metabolism in miners in deep coal mines]. AB - In a study into the condition of central hemodynamics and water-salt metabolism made in 85 essentially healthy underground miners working in a coal mine those miners greater than 40 years old exhibited an increase in the peripheral vascular resistance together with a decrease in the power of the left ventricle and a rise in energy expenditure on blood redirection, which fact led to lowering of both volume- and velocity-related parameters for hemodynamics. Worsening of hemodynamic indexes was noted to be going on parallel to changes in water-salt metabolism: workers beyond thirty years of age showed an augmentation of the level of Na+ and creatinine in blood plasma, those greater than 40 years old demonstrated a reduction in urine creatinine, in those beyond fifty years of age glomerular filtration rate was found to be on decrease with fractional excretion of Na+ being on the increase. PMID- 10822703 TI - [The efficacy of methods for the differentiated health rehabilitation of miners in risk groups for disease development]. AB - Of the miners in poor health, groups have been identified at high risk for development of respiratory, cardiovascular, and neural pathologies (n = 30). Treatment and prophylactic complexes were differentiated and implemented in the miners while on vacation in a sanatorium-preventorium setting. Efficacies are shown of differentiated methods for promotion of the health of those miners having been assigned to different groups at risk for development of pathologies of the respiratory organs, the cardiovascular, and the nervous system while on tariff vacation. The promotion-of-health courses have led to normalization of the immune and biochemical statuses in 68 percent of miners secondary to activation of chief links of cell-mediated, humoral immunity, factors of nonspecific bodily resistance and parameters for fat metabolism. PMID- 10822704 TI - [The state of specific functions in female workers subjected to exposure to epoxy resins and to epoxy resin-based polymeric materials during work activities (the data from a medical examination)]. AB - Our in-plant hygienic observations permitted ascertaining that female workers engaged in the manufacture of epoxy-based polymeric materials are exposed to a complex of adverse factors. Gynecological and extragenital pathologies are most prevalent as are disorders in specific functions of the female organism. It is considered necessary that a complex of practical, sanitary and hygienic, and treatment and prophylactic measures be implemented in order that disturbances in specific functions of the female organism might be prevented, values for the prevalence of gynecological and extragenital pathologies decreased. Hormonal colpocytoscopy is warranted in early diagnosis of the menstrual disorders. PMID- 10822705 TI - [The status of occupational morbidity in Ukraine today]. AB - Data are submitted on the pattern and levels of occupational morbidity in Ukraine in different branches of national economy, by forms of pathology, with those individuals presenting with occupational health problems having gotten distributed by sex, age, and service duration. The article addresses the issue of significance in the formation of occupational pathology and its levels of different factors and causes (economical, social, medical). The status of the occupational pathology service in the country is considered, with special emphasis being placed on inadequacy of detecting occupational diseases. PMID- 10822706 TI - [The scientific bases for determining the health losses of the affected population in catastrophic flooding]. AB - The article focuses on methodological approaches to the determination of sanitary losses among population in a catastrophic flood. An emergency is considered that can arise in the destruction of the water-development project assemblies. The following items were taken account of in working out of the above approaches: population density in the territories found to be in the zone of flood: notification and evacuation of the population from the zone of catastrophe in a timely fashion, the distance from the site of formation of the wave of breaking to the populated locality, the height and velocity of the wave of breaking, time of the year and day of the catastrophe. Scientific and methodological approaches and the mathematical model "Povin'" ("Flood") developed with the aid of such approaches permit analyzing the situation having presented itself, working out administrative decisions for the territorial public health services directing bodies to be taken in the current situation, instituting measures designed to eliminate medicosanitary effects of the catastrophic flood within the shortest time possible. PMID- 10822707 TI - [The principles of organizing and integrating valeological innovation centers in a market economy]. AB - Principles are submitted of organization and integration of valeological innovative centres of the recropark, recropolis, and health resortpolis type into the market economics. The above principles follow from modern scientific achievements, those in the fields of cybernetics, synergetics, and syntheloctics, from the development of society as a whole. PMID- 10822709 TI - Bond between gold alloy and ceramic in relation to the thickness of the oxide layer. AB - BACKGROUND: The chemical bonding between metal and ceramic is aided by non precious metal atoms and their oxides present on the metal surface. Aim of the study was to correlate metal-ceramic bonding with thickness oxide layer on a new gold alloy. METHODS: Thirty-two specimens were constructed in gold alloy and ceramic, with the aim of evaluating the degree (using shear test) and quality (using optical microscope) of bonding resistance between these materials, in relation to the thicknesses of the oxide layer produced on the metallic parts by means of different thermal procedures. RESULTS: Bond strength was practically the same with different thicknesses of the oxide layer. A slight reduction in the degree of bonding was observed in the non-oxidised models. CONCLUSIONS: For the gold-alloy here tested, the oxide layer seems to be not influent in the chemical metal-ceramic bond strength. PMID- 10822708 TI - Bioactivity of chitosan in dentistry. Preliminary data on chitosan-based cements. AB - BACKGROUND: The chemical association of chitosan with inorganic salts, such as calcium phosphate, finds a promising application in dentistry as room-temperature self-hardening cement. We present the physical, chemical and crystallographic characterization of newly-developed cements made of 1) calcium-phosphate and a chitosan gel obtained by acetic acid treatment, and 2) calcium phosphate and a chitosan gel obtained by ascorbic acid treatment. Both cements are self-hardening at room temperature. METHODS: The cements were characterized by X-ray diffractography, scanning electron microscopy and fluorine-selective electrode analysis. RESULTS: The chitosan-hydroxyapatite cements had hardness comparable to spongy bone and above that of PMMA. CONCLUSIONS: The cements are promising for application in endodontics and restorative dentistry. PMID- 10822710 TI - [Failures and surgical complications in oral implants. Follow-up of 327 osseointegrated implants]. AB - AIM: The study analyses the short- and long-term clinical results in 112 patients with total or partial edentulia of the jaw undergoing oral implantology in order to assess the type of surgical complications. METHODS: A total of 327 implants were inserted with an internal hexagonal screw, a diameter of between 3.25 and 5.5 mm and between 11 and 15 mm long. Each patient received three types of implant: overdentures, bridges supported by implants alone and bridges supported by implants and natural teeth. RESULTS: In line with other data reported in the literature, the failure rate at 7 years was 3%. CONCLUSIONS: After a general discussion of the possible surgical complications arising during implantology, the authors describe and analyse all the surgical complications that occurred in this series, regarding both implants alone, bridges supported by implants and natural teeth. PMID- 10822711 TI - [The chin hole. Position and correlation with other mandibular parameters]. AB - BACKGROUND: The present study analyzes the position of mental foramen, in order to research possible correlations with other mandibular parameters. METHODS: The study has been conducted on 76 dry mandibles, for each mandible we have measured the exact position of the mental forum respect the other mandibular parameters. Both linear and angular relations and misurations have been made. RESULTS AND CONCLUSIONS: The results obtained confirm that the mental foramen maintains its position by increasing GoGn. The results did not show particularly significant aspects in comparison with the literature on the subject, thus confirming other authors' opinion. PMID- 10822712 TI - Non neoplastic tongue diseases. An epidemiological investigation and diagnostic criteria. AB - BACKGROUND: Few clinical-epidemiological data regarding tongue diseases were showed in recent literature. The aim of this study was to evaluate the prevalence of non neoplastic tongue pathologies, and to perform an epidemiological, clinical and etiopathogenetic comparable data system on Non-Neoplastic Glossitis (NNG). METHODS: A total of 215 subjects (90 males and 125 females, age range: 6-72 years) have been examined) at the Dental Clinic of the University of Brescia over a period of 2 years. From this group, patients with tongue non neoplastic lesions were selected. Each selected patient with NNG was examined, following the Diagnostic Protocol of the "Department of Oral Pathology and Medicine" of the Dental Clinic of Brescia University. RESULTS: 84 cases of NNG (39%) were observed from January 1997 to October 1998. CONCLUSIONS: The selected group of patients with NNG has been stratified following clinical and etiological criteria, and the results discussed, emphasizing the importance of careful and correct examination of the tongue, in order to bring to light morphological and pathological changes often neglected or misdiagnosed. One should never exclude, moreover the possibility of malignant evolution of some lingual lesions requiring a close follow-up. PMID- 10822713 TI - [Surgical wound healing in oral surgery]. AB - BACKGROUND: Aim of this study is to improve the in vivo knowledge about the real effects and the physiologic repair by a detached stitches silk suture in the 8th zone. METHODS: The study has been carried out on 44 patients submitted to surgical extraction of the impacted 8th (19 upper and 25 lower) teeth. RESULTS: Clinical and histological controls have demonstrated the usefulness of this suture procedure, since it gives a good histological reaction and in easy and safe. CONCLUSIONS: Histological and clinical results have confirmed the usefulness of silk suture, presenting however the inconvenience of stitches removal and bacterial plaque on the surgical wound. PMID- 10822714 TI - [Orodental management in patients with malignant hematologic diseases who are waiting for bone marrow transplantation]. AB - BACKGROUND: This study aims to identify the main oral lesions caused by the use of chemotherapeutic agents pre- and post-transplant in malignant hemopathologies and to highlight the importance of dental prevention during this treatment in order to reduce and, if possible, avoid the resulting local and systemic complications. METHODS: Twenty-five patients were controlled, monitored and prepared to undergo the necessary chemo- and radiotherapy, as well as bone marrow transplantation in the majority of cases under the best possible oral conditions. RESULTS: The most commonly detected secondary lesions were reactions to cyclophosphamide and/or methotrexate. In general, this mucositis appeared 5-7 days after treatment and continued for 1-2 weeks before regressing without residue. Another post-radiotherapeutic complication observed was xerostomy. CONCLUSIONS: Lastly, it is important to underline that patients receiving correct dental infection preventive treatment showed fewer tertiary lesions than those who did not undergo the same procedure. PMID- 10822716 TI - [Basaloid squamous carcinoma of the tongue. Clinico-anatomical study of a case]. AB - The basaloid squamous carcinoma was first described in 1986. It is a rare tumor with particular morphologic and biologic features that separate it from the more common squamous cell carcinoma of the head and neck region. A case with histologic diagnosis of basaloid squamous carcinoma of the tongue treated only with radiotherapy, is reported. PMID- 10822715 TI - [Radiopaque lesions of the jaws]. AB - This paper provides an overview of radiopaque lesions of the jaws after having outlined the various criteria for classification; depending on their etiology. Those lesions are generally separated into lesions with an unknown cause and lesions with a known etiology (phlogistic, dysplastic-dystrophic, tumoral). The authors give a general description of the clinical and radiographic symptoms of the main lesions. In many cases, with the exception of asymptomatic lesions with an unknown etiology, the patient presents a variable swelling that deforms the bone in question, resulting in asymmetry of the face and/or, depending on the site, nasal obstruction, mastication and speech disorders, exophthalmos with or without diplopia. The radiographic aspect of lesions and the associated laboratory and clinical findings often enable a diagnosis to be reached, but in doubtful cases diagnosis can only be confirmed by histological analysis. PMID- 10822717 TI - [New suture techniques in oral surgery. Description and comparison with traditional sutures]. AB - The study aimed to evaluate the healing of intraoral surgical wounds sutured with alpha-cyanoacrylate and silk in clinical and histological terms. Forty-four patients were checked 7 and 14 days after surgery. An incisional biopsy was made at the level of the discharge cut and histological tests were performed on the fragment. On the basis of the results, the authors affirm that in some areas (upper eighths), alpha-cyanoacrylate plays a useful role and simplifies the suture technique. On the contrary, this material was not sufficiently reliable in areas subject to excessive traction (lower eighths). PMID- 10822718 TI - [Nephrology: 21st century]. PMID- 10822719 TI - [Kidney transplantation from living donor: a real therapeutic option]. PMID- 10822720 TI - Draft protocol on transplantation of organs and tissues of human origin. Council of Europe. PMID- 10822721 TI - [Mutational analysis of the PKD1 and PKD2 (type 1 and 2 dominant autosomal polycystic kidney) genes]. AB - Autosomal dominant polycystic kidney disease (ADPKD) is the most common hereditary kidney disease. It is caused by mutations in at least two different genes: PKD1 and PKD2. The study of mutations in these genes is very difficult nowadays. In this study we have analyzed the non reiterated region of the PKD1 gene and all the exons and intron exon boundaries of the PKD2 gene. The technique used to study these genes have been single strand conformation analysis and heteroduplex. We have found 25 differences within the DNA sequence of the PKD1 gene with respect to the published sequence. Seven of these changes correspond to nonsense, missense, frameshifting and splicing mutations. The rest of changes correspond to polymorphisms or rare DNA variants. In the PKD2 gene we have identified 8 new mutations and one polymorphism. Six of these mutations are frameshifting, one is missense and the other one is a large deletion of the PKD2 gene. The rate of mutation detection within the PKD1 gene has been 4% and the rate for PKD2 has been 100%. We have not observed any correlation between genotype and phenotype either in the PKD1 nor in the PKD2 gene. The mutation analysis of ADPKD genes is very difficult, specially for the PKD1 gene. The rate of mutation detection is higher in the PKD2 gene but the global efficacy of the technique is very low as PKD2 represents only 15% of ADPKD patients. Nowadays linkage analysis is still the most useful technique for the molecular diagnosis of ADPKD patients. PMID- 10822722 TI - [Apolipoprotein C-II and C-III anomalies in normolipemic and hyperlipemic patients with chronic kidney failure]. AB - The high incidence of arteriosclerotic disease in patients with chronic renal failure seems to be due to certain peculiarities in their lipid metabolism. These are principally a disorder in the transportation of lipoproteins and a concomitant defect in triglyceride metabolism causing an accumulation of triglyceride-rich-lipoproteins which predispose to atherosclerosis. We studied the disturbances in concentration of apolipoproteins, notably Apo C-II and C-III, which modulate the activity of lipoprotein lipase (LPL), in patients with chronic renal failure (CRF) without replacement therapy and in hemodialysis patients with and without hyperlipidemia. LPL hydrolyses triglycerides in the lipoprotein triglyceride (LPRTG) core. The main lipid parameters were measured in 4 groups of normolipidemic and hyperlipidemic patients with and without CRF in comparison with healthy controls. We found that the lipolytic activity index (A-I/C-III) was decreased, and Apo C-III levels were increased, in patients with CRF and patients on HD, including normolipidemic patients. We conclude that high Apo C-III levels are found in uremic patients before starting dialysis and do not change during dialysis treatment. This increase could be one of the initial causes of impaired triglyceride catabolism and LPRTG accumulation even in normolipidemic patients with CRF and may be one explanation of the high mortality from cardiovascular disease in these patients. PMID- 10822723 TI - [Experience of a renal function test unit]. AB - The activity developed during one year in a Renal Function Tests Unit was studied prospectively. Six hundred pharmacological tests (50 tests per month) were done to 453 patients, children and adults. Abnormal tests results were found in 38.4% of the patients. 49.7% of the tests were performed in patients affected with nephrolithiasis or carriers of metabolic anomalies predisposing to renal stones, mainly idiopathic hypercalciuria. The tests were also frequently done to other group of patients affected with urinary tract infection, mitochondrial disease, vesicoureteral reflux, osteoporosis and primary Sjogren's syndrome. The urinary concentrating capacity after desmopressin (37.2%) was the most frequent performed isolated test, although the group of tests to study the renal acidification capacity (58.5%) were the most frequent requested ones. PMID- 10822724 TI - [Interaction of the mechanisms of beta 2-microglobulin convection, diffusion, and adsorption in line hemodiafiltration]. AB - The in vivo contribution of diffusion, convection ad adsorption to beta 2 microglobulin (beta 2-m) elimination by hemodiafiltration (HDF) was investigated. 11 patients (8M/3W), with a mean age of 59 +/- 10 years and weighing 62.7 +/- 8.7 kg were studied. A 1.89 m2 polysulphone membrane was used in 180 min postdilution HDF. Samples at blood inlet (bi), blood autlet (bo), dialysate outlet (do) and ultrafiltrate (uf) were taken to determine beta 2-m concentrations at 30 and 150 min. Rates of flow (Q, ml(min) prescribed were: infusion, Qinf = 103.6 +/- 12.3, Quf = 14.6 +/- 4.0 y Qb = 465 +/- 5.0. Effective Qbi was automatically measured by the machine and Qdo = 800 + Quf. The removed beta 2-m mass (M, mg/min) was obtained by multiplying rates of flow (Q, L/min) by beta 2-m concentrations (mg/L) at each sampling point. From mass balance, we calculated the mass of beta 2-m removed (mg/min) by adsorption 0.23 +/- 0.2, by convection 0.7 +/- 0.3 and by diffusion 1.0 +/- 0.4, at 30 min. At 150 min, the beta 2-m mass removed was -0.06 +/- 0.1 by adsorption 0.4 +/- 0.1 by convection and 0.3 +/- 0.1 by diffusion. In HDF, these beta 2-m eliminating mechanisms play a variable role throughout the session. The more significant conclusion is that diffusion of beta 2-m with a synthetic "open" membrane is an important method of removing beta 2-m, comparable to convection over the whole procedure. That result explain the relative efficacy of beta 2-m clearance by HDF convection, and also explain why isolated diffusion is an efficient mechanism for beta 2-m removal by high-flux hemodialysis. PMID- 10822725 TI - [Changes in the anion gap in patients undergoing hemodiafiltration]. AB - The serum anion gap (AG) is a calculated value defined as the difference between the sum of sodium and potassium and the sum of chloride and bicarbonate concentrations. Thus, the anion gap is equal to the unmeasured cations minus the unmeasured anions (UA). To evaluate the AG changes during HDF-on line, we studied 20 patients treated with this technique. Blood pH, HCO3, NA, K, Cl, albumin, phosphorus, urea, creatinine and lactate were determined pre and post-HDF. The AG, negative charger of serum albumin (CAA) and phosphate (CAP) were computed by equations. AG decreased during HDF from 23.1 +/- 3.4 mEq/l to 17.3 +/- 3.6 mEq/l (p < 0.001). The CAA rose from 10.9 +/- 0.8 to 12.3 +/- 1.7 mEq/l (p < 0.001). The CAP and lactate fell significantly during HDF (p < 0.001 and 0.05 respectively). Other unmeasured anions (UA) decreased from 7.9 +/- 3.0 to 2.4 +/- 2.7 mEq/l (p < 0.001). The CAA contributed 47.7 +/- 6.5% and 73.01 +/- 12.7% to the pre and post-HDF serum anion gap respectively. The CAP accounted for 12.4 +/- 3.4% and 8.6 +/- 1.8%, lactate 6.4 +/- 3.9% and 6.0 +/- 3.0% and UA for 33.2 +/- 7.7% and 12.2 +/- 13.6% of the anion gap pre and post-HD respectively. AG and UA correlated significantly with blood urea pre-HDF and urea generation. The increase in serum albumin and pH can mask an decreased concentration of unmeasured anions in patients treated with HDF on-line. An adjusted anion gap without effect of CAA and CAP can be obtained. With the help of this adjustments the changes in some undetermined anions organic and inorganic (sulphate and others in renal failure) can be calculated. PMID- 10822727 TI - [Anti-glomerular basement membrane disease: a new disease causing fever of unknown origin?]. AB - Antiglomerular basement membrane disease is an autoimmune disorder characterized by the presence of antibodies directed against glomerular basement membrane. Pyrexia of unknown origin (PUO) is defined as temperatures higher than 38.3 degrees C on several occasions, with a duration of more than 3 weeks, and failure to reach a diagnosis despite 1 week of in-patient investigation. There is a large list of causes of PUO including infections, malignancies and autoimmune diseases, but antiglomerular basement disease has not been described as a cause. We present the first case of antiglomerular basement disease which presented with PUO. PMID- 10822726 TI - [Relationship between the 1993 Daugirdas Kt/V method and other methods to calculate the dialysis dose]. AB - INTRODUCTION: There are several shortcut formulas to calculate Kt/V, the most widely used because of its simplicity is the logarithmic formula proposed by Lowrie in 1983. The DOQI report recommends use of the formula proposed by Daugirdas's in 1993 to estimate the Kt/V, it must be > or = 1.2. The aim of the present work was to analyse the concordance between the second generation Daugirdas's formula and the other shortcut formulas. MATERIAL AND METHODS: In 208 hemodialysis sessions performed on 61 patients, Kt/V was estimated by thirteen formulas: 10 single-pool modeling and three double-pool modeling. RESULTS: The Kt/V values obtained by Daugirdas's formula was different from those obtained with the other single-pool based formulas (p < 0.01). When the dialysis sessions were classified in 4 groups according to the Kt/V values calculated by Daugirdas's formula, most of the single-pool formulas gave Kt/V values statistically different from those given by Daugirda's formula in all ranges examined. The concordance among Kt/V calculated by every one of the single-pool formulas and Daugirdas's formula was variable. The highest agreement was with Keshaviah's and Lowrie's 1992 formulas and the lowest with Calzavare's and Lowrie's 1983 formulas. The linear multivariate analysis showed that the two factors which influenced the concordance were the Kt/V value obtained by the Daugirdas's formula, and the quotient between the postdialysis body weight and the ultrafiltration rate. Despite the weak concordance with Daugirdas's formula, the Kt/V obtained by Lowrie's 1983 formula and the urea reduction ratio (URR) are still useful tools to monitor the adequacy of dialysis: a Kt/V > 1.065 calculated by the Lowrie's 1983 formula and a PRV > 65% are equivalent to a Kt/V > or = 1.2 calculated by Daugirdas's formula. CONCLUSIONS: The shortcut formulas used to calculate Kt/V give different results with high inter-method variability. When Kt/V is reported it is absolutely necessary to indicate the formula used to calculate it. The Kt/V obtained by Lowrie's 1983 formula and the URR are useful tools to monitor dialysis adequacy. PMID- 10822728 TI - [Hypophosphatemia as diagnostic clue in multiple myeloma]. PMID- 10822729 TI - [Rhabdomyolysis caused by carnitine palmitoyltransferase deficiency. Prevention of acute kidney failure]. PMID- 10822730 TI - [Posterior leukoencephalopathy caused by erythropoietin]. PMID- 10822731 TI - [Renal, hepatic, and neurologic toxicity caused by NSAIDs]. PMID- 10822732 TI - [Introduction to the 6th International Symposium of the "Reina Sofia" Institute of Nephrologic Research]. PMID- 10822733 TI - Gene therapy in the vasculature and the kidney. AB - Gene therapy has many potential applications for the treatment of vascular disease. Though a number of tough problems remain to be solved, the potential specificity with which an almost limitless number of mechanisms could be targeted, and the success that has been achieved in animal models in vivo make it likely that we will see further rapid expansion of this technology, and therapeutic use of gene therapy in humans in the future. PMID- 10822734 TI - [The NFAT transcription factor family as immunosuppression target]. PMID- 10822735 TI - [Cyclosporin A produces intracellular superoxide anion in endothelium]. PMID- 10822736 TI - [Regulator role of apoptosis in peritoneal cellularity]. PMID- 10822737 TI - [Type I early failure of peritoneal ultrafiltration (UF)]. PMID- 10822738 TI - [Automatic peritoneal dialysis]. PMID- 10822739 TI - [Clinical implications of peritoneal transport]. PMID- 10822740 TI - History and role of Psychology of Addictive Behaviors. PMID- 10822741 TI - Rediscovering fire: small interventions, large effects. AB - Unexpected findings are often the spark for new discoveries and theories. A puzzle emerged from a series of unanticipated findings over 3 decades, indicating that for problem drinkers (a) relatively brief interventions can trigger significant change, (b) increasing the intensity of treatment does not consistently improve outcome, (c) therapist empathy can be a potent predictor of client change, and (d) a single empathic counseling session can substantially enhance the outcome of subsequent treatment. These phenomena are considered in light of other findings in the addictions-treatment-outcome literature. There is, at present, no cogent explanation for the efficacy of brief interventions. An ancient construct is explored as one possible factor in how some brief encounters may exert large effects in human change. PMID- 10822742 TI - Familial risk for alcoholism and self-reported psychopathology. AB - Three groups of young men varying in familial alcoholism risk (high density, high risk [HDHR]; low density, high risk [LDHR]; and low risk [LR]) were compared on the 11 clinical scales of the Personality Assessment Inventory. Significant group differences were found on 9 scales, with scores of the HDHR group exceeding those of the other 2 groups. No differences were found between the LDHR and LR groups. When the proportion with pathological scores per scale was examined, significant group differences were still revealed on 7 scales. The HDHR group exceeded the other 2 groups, but the LDHR group also exceeded the LR group on several scales. These findings support the need to more finely characterize familial alcoholism risk than is provided by the typical high-risk-low-risk dichotomy. Finally, statistically controlling for normal variations in response style reduced the number of group differences, although the same patterns persisted. PMID- 10822743 TI - Assessing and training recognition of intoxication by university students. AB - Forty-five undergraduate volunteers attended either a 2-session recognition of intoxication training program or a 2-session attention-placebo control group. Stimulus videotapes portrayed male and female university students (targets) performing various tasks both before drinking and again after consumption of 3 to 5 standard drinks. Participants' accuracy in recognizing whether videotaped targets had or had not been drinking (target status) was assessed 1 week posttraining and 2 months posttraining. At the 1-week follow-up assessment, participants in the training program had a higher mean number of accurate ratings of target status compared with the attention-placebo control group. In addition, a larger proportion of training-group participants expressed an overall preference to rate a target as not having been drinking if they were unsure of target status. These effects were not maintained at the 2-month follow-up assessment. PMID- 10822744 TI - Preventing alcohol misuse: the impact of refusal skills and norms. AB - The purpose of this study was to determine the extent to which refusal skills and norm setting mediated the impact of a school-based prevention program from the Alcohol Misuse Prevention Study (AMPS) on adolescent alcohol overindulgence. The AMPS is a randomized, pre-post, experimental-control study. Respondents in the present study included 6th through 10th graders (ns ranged from 232 to 371). Structural equation modeling analyses using EQS indicated that norm setting mediated the effect of the intervention on alcohol overindulgence at the 7th through the 8th grade and at the 8th through the 10th grade. In contrast, although the prevention program served to increase refusal skills, refusal skills did not mediate the effect of the program on alcohol misuse. PMID- 10822745 TI - Behavior change patterns and strategies distinguishing moderation drinking and abstinence during the natural resolution of alcohol problems without treatment. AB - Behavior change patterns and strategies involved in natural resolutions that resulted in stable moderation drinking or abstinence were investigated, using untreated problem drinkers with different drinking statuses. Participants' drinking practices and problems, resolution patterns, behavior-change strategies, and barriers to help seeking were assessed during structured interviews. Collaterals verified participants' reports. Most abstinent resolutions were initiated abruptly. Moderation resolutions were achieved more gradually and entailed changes in drinking practices like those emphasized in behavioral self control treatments. Participants' desire to solve their own problem and concerns about available interventions deterred help seeking, even though help was widely available. These data suggest that variability exists in how drinking problems are resolved and that interventions should support the several successful resolution patterns. PMID- 10822746 TI - Temperament and antisocial behavior in preadolescent boys with or without a family history of a substance use disorder. AB - The goals of this study were to determine the relations between different dimensions of temperament, and their interactions, with antisocial behavior (ASB) in 351 preadolescent boys with (n = 175) or without (n = 176) a family history of a substance use disorder (SUD) and to determine whether these relations are moderated by a family history of SUD. Participants were administered the Revised Dimensions of Temperament Survey (DOTS-R) and multiple measures of ASB. Factor analysis reduced the DOTS-R subscales into three factors: Rhythmicity, Behavioral Regulation, and Positive Affectivity. Results indicated that above and beyond the effects of age and socioeconomic status, low rhythmicity, low behavioral regulation, and low positive affectivity, as well as some of their higher order interactive effects, are important indicators of different types of ASB and, in some cases, only in boys with a family history of SUD. PMID- 10822747 TI - Contingency management for accurate predictions of urinalysis test results and lack of correspondence with self-reported drug use among polydrug abusers. AB - Contingency management procedures have proven effective in the treatment of drug dependent patients. These procedures, however, often require frequent urine testing, which is too costly for community treatment programs. To make urine testing procedures more cost effective, the feasibility of reinforcing accurate predictions of urine drug screen (UDS) results was evaluated. Participants made extremely accurate UDS predictions, particularly when they made drug-positive predictions, regardless of whether predictions were reinforced. However, self reports of recent drug use had poor correspondence with predictions of UDS results. Results suggested that if programs only tested samples predicted to be drug free, considerable cost savings could be incurred. Further research is needed to determine if validity would be enhanced by using a proportion of costs saved to provide nominal reinforcement when samples were verified to be drug free. PMID- 10822748 TI - Absentminded lapses during smoking cessation. AB - Although it is widely believed that drug cravings are responsible for drug use and relapse, S. T. Tiffany (1990) has proposed a cognitive model in which drug use is triggered not by craving but by the cuing of automatized action plans. The purpose of this study was to examine the lapse episodes from an ecological momentary assessment (EMA) study of smokers attempting to quit for evidence of automatic, or absentminded, lapses with slight or no urges to smoke, in keeping with S. T. Tiffany's (1990) model. Qualitative analysis of 270 EMA reports made by 41 smokers during the first 14 days of quitting identified 15 (6%) absentminded lapses. Quantitative urge levels were significantly lower during absentminded lapses compared with nonabsentminded lapses; however, urges were not very low. Results indicate that absentminded lapses may occur but are probably relatively rare. PMID- 10822749 TI - Relations between dietary restraint and patterns of alcohol use in young adult women. AB - The present study examined relations between dietary restraint and self-reported patterns of alcohol use, including separate assessment of quantity and frequency of alcohol consumption. One hundred seventy-six female university undergraduates completed the Restraint Scale (RS) and measures of their usual quantity and frequency of alcohol consumption over the past year. Quantity and frequency self reports were scored separately and were also used to calculate 3 additional drinking variables: a composite weekly alcohol consumption score (drinks per week), a binge drinking categorical variable (where participants were classified as either binge drinkers or non-binge drinkers), and a yearly excessive drinking score (number of times in the past year that each participant consumed at least 4 alcoholic beverages per drinking occasion). RS scores were significantly positively correlated with scores on 4 of the 5 drinking behavior measures (i.e., quantity, drinks per week, binge drinking, and yearly excessive drinking, but not frequency). Thus, chronic dieting appears to be related to a relatively heavy drinking pattern that can be characterized as potentially risky, due to its established associations with adverse health and social consequences. PMID- 10822750 TI - A comparison of models of drinking motives in a university sample. AB - Previous research has suggested the presence of several alternative factorial models for the assessment of drinking motives. In the present study, confirmatory factor analysis was used to assess the factor structure of the Drinking Motives Questionnaire (M. L. Cooper, 1994) in a college sample. The results indicate that a 4-factor model that includes the dimensions of social rewards, affect enhancement, coping, and conformity motives fits the data significantly better than 2- and 3-factor models. Furthermore, the 4-factor model fits equally well for men and women. PMID- 10822751 TI - [Lupus anticoagulant: complete automation (ACL-Futura) and diagnostic algorithm]. AB - PURPOSE: The technical and clinical evaluation of a new laboratory profile with six tests, for the realization of the assay called "lupic anticoagulant" with the help of ACL-Futura analyzer (Intrumentation Laboratory). RESULTS: The within between day imprecision of the tests that compose the profile: APTT-Diluted, APTT D Mix, LacScreen, LacScreen Mix, LacConfirm and LacConfirm Mix, are between 2.87% 11.61% with controls, this imprecision is lowest with patients. A study of bilirrubin and lipemia interferences is presented. The practicability study present the technical, time consuming difficulty, and shows that the cost of the screening test is about 1136 ptas (6.83 euros) and with the confirmative test is 2766 ptas (16.62 euros). The clinical study describes our preliminary results with the application of this new profile for almost 2 years and six clinical cases are presented. CONCLUSIONS: The good technical and clinical results of the evaluation of the new profile proposed to detect the positive lupic anticoagulant, in addition to the fully automated assay with the ACL-Futura analyzer, validate the whole method to resolve the increased demand of these parameters. PMID- 10822752 TI - [No ELISA detectable alterations in immunogenicity following dry-hear treatment (72 hours at 80 degrees C) of FANHDI]. AB - OBJECTIVE: Neoantigen formation during heat treatment (HT) of factor VIII:von Willebrand Factor (FVIII:vWF) concentrates may induce an immune response against the modified protein, which may also affect the native protein. We present a comparative in vitro study on the immunogenicity of a dual virally inactivated (solvent-detergent and 80 degrees C 72 hours) high purity FVIII:vWF concentrate (Fanhdi) versus the same product without heat treatment. MATERIAL AND METHODS: For this purpose rabbit antisera were prepared using both Fanhdi and the same product from which the human albumin, used as stabilizer, had been removed (these were both HT products). Also, antisera were prepared against the same products made without the dry-heat treatment step (non-HT products). Antisera were analysed by Elisa. Mixtures of antisera with increasing amounts of product (incubation-absorption in liquid phase) were assayed in plates coated with HT and non-HT products. RESULTS: The binding of antibodies against HT products to ELISA plates coated with HT products, could be blocked (in a saturable manner) with non HT products, following liquid phase incubation. These results strongly suggest the absence of neonantigens. Furthermore, the binding of antibodies against non HT products to ELISA plates coated with non-HT products, could be blocked (also in a saturable manner) with HT products. This result indicates that there is no epitope loss. CONCLUSIONS: The data obtained in these studies suggest that the heat treatment of viral inactivation as applied in Fanhdi, does not give rise to any major alteration in immunogenicity of the product. The data from clinical and drug surveillance studies carried out with Fanhdi do not show any indication of an increase in the frequency of inhibitors. PMID- 10822753 TI - [Etiological study and diagnosis of anemia in adults over 60 years of age]. AB - PURPOSE: Anaemia especially iron deficiency anaemia (IDA) is a worldwide health problem and the most frequent nutritional lack in developing countries. The epidemiology of anaemia in hospitalized patients in Internal Wards is not well known. PATIENTS AND METHODS: On a retrospective basis we studied 105 patients with haemoglobin levels below 115 g/L. Symptoms, type of anaemia, causing disease, diagnostic procedures, pathologic findings and transfusional schedule were analyzed. RESULTS: Mean haemoglobin was 77.8 (SD 17.9) g/L. Anaemia degree was deeper in women (p < 0.05) and IDA comparatively with chronic disease (CDA) (p < 0.01) and it did not show relation with therapeutic agents which potentially induce anaemia. IDA was the most frequent followed by haemorrhagic anaemia (HA) and CDA. The diagnostic procedure which discovered an underlying disease in most of the cases was upper digestive tract endoscopy, and CDA needed the highest number of diagnostic procedures. There was not relationship between the sort of anaemia and symptoms due to upper digestive tract and endoscopic findings, however low digestive tract symptoms (bleeding and abnormal finger rectal examination) and pathologic findings in colonoscopy had a straight relation (p < 0.05). CONCLUSIONS: IDA is the anaemia most frequently diagnosed in an Internal Medicine Department followed by HA and ACD. Clinical symptoms and the type of anaemia have poor correlation. In IDA upper digestive endoscopy as well as lower digestive tract examination is mandatory. We propose colonoscopy in patients aged more than 50-years whereas barium enema could be employed in younger people. PMID- 10822754 TI - [Detection of hypovolemia according to Weinreb's criteria in 30 patients previously diagnosed with relative polycythemia]. AB - PURPOSES: To apply the criteria for classifying the normo- or hypovolaemic state to the blood volume of patients previously diagnosed of relative polycythaemia (RP). PATIENTS AND METHODS: The blood volume of 30 patients previously diagnosed of RP was chosen at random for analysis. Directly measured cell volume (CV) was used to calculate both the total blood volume (WBV) and the plasma volume (PV) of each case expressed as mL and mL/Kg, respectively. WBV was estimated according to Nadler's predictive formula, CV and PV being deductive values expressed in mL. Corrected venous haematocrit (CVH) CV and PV measured as mL/Kg were compared with respective reference intervals of the laboratory (RIL). Weinreb's criteria, modified in accordance to RIL, were used to classify blood volume as normo- or hipovolaemic. RESULTS: Of the 30 patients, 27 were men and 3 women. Mean CVH was 0.51 L/L (range 0.45-0.58 [L/L]). In mL/Kg, 80% of the patients had normal CV with reduced PV, 10% showed normal CV and PV, and the remainders had increased CV with normal PV in 3.3% and decreased PV in 6.7%. Hypovolaemic state was found in 100% of the patients. CONCLUSIONS: Male sex was confirmed as predominant with regard to RP patients. Mean CVH was lower than that of absolute polycythaemia patients, and its wide range stresses the importance of blood volume in the diagnosis of this type of patients, even though reduced PV with normal CV, according to RIL in mL/Kg, are not present. The previous diagnosis was corroborated, hypovolaemic stated being confirmed in all cases at the moment of the study, so more accurate diagnosis and adequate therapeutic indications could be granted, in accordance to the patient's status. PMID- 10822755 TI - [Rh (D) alloimmunization and pregnancy. Analysis of the causes after prophylaxis introduction]. AB - PURPOSE: Prenatal and postnatal prophylaxis of the Rh (D) haemolytic disease of the newborn have clearly reduced the number of cases but still there are alloimmunizations. PATIENTS AND METHODS: All cases detected in our Hospital in the last 24 years have been reviewed and possible causes analyzed. RESULTS: From a total of 10,332 deliveries in Rh (D) negative women we have detected 114 anti-D in 86 women. In 74 women anti-D was the only antibody and in 12 there were more antibodies. Data were managed in 3-year periods and we see a progressive decrease in the incidence of alloimmunization with a minimum of 0.03 per 1000 pregnancies in the period 89-91 and a posterior progression to an incidence of 0.12 in the last 3-year period 95-97. The causes were: pregnancies before 1970 in 31, incorrect prophylaxis in 12, despite a correct prophylaxis in 6, previous pregnancies without complete information about the prophylaxis in 13, previous transfusion in 6, previous pregnancies or transfusion in 8 and indetermined in 10. CONCLUSION: It is desirable to reduce at minimum the number of Rh (D) alloimmunizations by strictly following the prophylaxis protocols. PMID- 10822756 TI - [Anti-A antibodies and bacterial contamination of platelet concentrates]. AB - OBJECTIVES: The possible ABO group antibodies protective function against several infections has been classically described. We analyze the platelet concentrates (PC) bacterial control results and their ABO antibodies. MATERIAL AND METHODS: We studied 245 outdated PCs (> 5 days). The samples were sterilely collected for adequate microbiological investigation studies on sheep-blood agar plates. If bacterial growth is found, the microbiological identification is performed on the basis of standard tests, the specific anti-biotype being achieved by disk diffusion method on Mueller-Hinton agar plates, and the red cell concentrate was analyzed. RESULTS: Bacterial growth by negative coagulase Staphylococcus was found in 10 PCs (4.1%; CI95%; 1.97-7.37). The contaminated PCs lacked natural anti-A antibodies. There were no statistical differences when we analyzed the PC's age, colour or blood group. COMMENTS: The anti-A antibodies may be a protective factor versus PCs contamination caused by resident bacteria. PMID- 10822757 TI - [Escherichia coli L-asparaginase induces phosphorylation of endogenous polypeptides in human immune cells]. AB - PURPOSE: To detect patterns of endogenous polypeptide phosphorylation in monocyte, lymphocyte, and polymorphonuclear leukocyte populations, induced by the products of the catalytic action of L-asparaginase (EcA). MATERIALS AND METHODS: Monocytes, polymorphonuclear cells and lymphocytes were isolated from heparinized blood from healthy, voluntary donors. The samples were incubated in 0.4 mCi/ml of [gamma-32P]H3PO4, with: 1 microgram/microliter of EcA, EcA and the substrate or with the products of EcA's catalytic activity: NH4+ and aspartate. The cells were lysated and electrophoresed using denaturing polyacrylamide gels that were then exposed on radiographic plates. The levels of polypeptide phosphorylation were quantified by computer densitometric analysis. RESULTS: The autoradiographs and the densitometric quantification of the electrophoretic profiles of monocytes, polymorphonuclear leukocytes, and lymphocytes revealed an increase in polypeptide phosphorylation when the cells were incubated with the enzyme and its substrate, ammonium and aspartate, or ammonium, which demonstrates that the NH4+ triggers intracellular phosphotransferase activity. A 58 kDa phosphoprotein outstood, it being common to the three cell populations studied. There were also specific phosphorylable polypeptides in monocytes, polymorphonuclear leukocytes, and lymphocytes. CONCLUSIONS: Escherichia coli L-asparaginase, binds the plasma membrane in normal human immune cells, catalyzing the L-asparagine substrate. The products of its activity: aspartate and NH4+ modify the extracellular environment, particularly the latter since it could diffuse into the cytosol and modify the pH, which would activate signal transduction pathways associated with the phosphorylation of substrates. PMID- 10822758 TI - [Pre and post-operative autotransfusion. A comparative study of hematology, biochemistry and red cell metabolism in pre-donated blood and blood from post operative surgical drainage]. AB - PURPOSE: The widespread use of aggressive surgical procedures, along with the increasing incidence of traffic accidents, has raised the necessity of homologous blood beyond the supplies of blood banks. This fact, plus the risks of homologous transfusion and the costs of blood bank maintenance, has prompted the advance of blood saving procedures such as autotransfusion, both in the pre-deposit (preoperative autotransfusion, POA) and the surgical drainage reinfusion (postoperative autotransfusion, SDR) modalities. As there is some controversy about the use of one or the other of the above procedures, the purposes of this study were: 1) to analyse the haematological and biochemical characteristics of blood, both pre-donated and stored at 4 degrees C for 4 weeks, and that recovered from surgical drains, from patients subjected to programmed orthopaedic or heart surgery; 2) to assess the metabolic and functional state of red cells attained from that blood, and 3) to compare the results achieved. MATERIAL AND METHODS: The following data were examined: red cell count, haematocrit, haemoglobin, red cell indicates, white cell count, platelet count, free plasma haemoglobin, red cell morphology, glucose, cholesterol, triglycerides, phospholipids, serum proteins and their fractions, ions, histamine, red cell glucose and amino acid transport, and ATP and 2,3-DPG content. RESULTS: The red cells, haemoglobin and haematocrit concentrations in POA blood did not show significant variations during the storage for 4 weeks and their values were significantly higher than found in drained blood. The biochemical values showed heterogeneous variations. Glucose and amino acid uptake by red cells of POA blood slightly decreased in the first 2 weeks of storage but always less than measured in SDR red cells. In POA blood it was noticed a progressive decrease in intra-erythrocytic ATP and 2,3 DPG, those levels being normal in SDR. DISCUSSION: In spite of lower haematocrit and haemoglobin but higher free plasma haemoglobin content, drained blood had higher ATP and 2,3-DPG concentration than pre-deposit, stored blood. Drained blood showed also less ion alterations and probably a lesser immunosuppressor capability. Thus, postoperative blood recovery seems a good source of red cell, with high oxygen transport power, and so, alone or in combination with pre donated blood, it may contribute to reduce the necessities for homologous blood and decrease its risks. PMID- 10822759 TI - [An analysis of transfusion in adult surgery]. AB - PURPOSE: To analyze the transfusion rate of adult transfused patients undergoing standard curative surgery made in operating room (not ambulatory). PATIENTS AND METHODS: Data from discharge reports of patients having surgery during 1996, codified according to the International Diseases Classification (ICE-9-MC). Age, sex, diagnosis, procedures (including transfusions), were analysed along with the surgical procedures and if the patient was subsequently admitted. RESULTS: 11,673 adult patients had surgery, of whom 890 underwent transfusion (7.6%). The transfusion rate was greater in males, in patients over 60 years old, in patients with several surgical procedures simultaneously, in patients who were subsequently admitted, and in patients with surgery procedures of pancreas, spleen, dorsolumbar column, liver, pharynx, hip and stomach. CONCLUSIONS: Given the variability in the blood transfusion practices among the hospitals, to know the transfusion rates of each clinic and surgical situation would permit a better planning of: the surgical blood transfusions, the autologous blood transfusion program, the criteria on crossmatching tests, as well as to increase the information to the patient in the surgical Informed Consent. PMID- 10822760 TI - [Treatment of leukemia relapsed after allogenic bone marrow transplantation with donor lymphocyte infusion: report of 11 cases]. AB - PURPOSE: Donor leukocyte infusions (DLI) are useful for treating leukaemic relapse after allogeneic bone marrow transplantation (BMT). We reviewed our experience with eleven patients who received DLI between 1995 and 1997. PATIENTS AND METHODS: The diagnoses prior to DLI were: chronic myeloid leukaemia (CML) in chronic phase (CP) (two patients) or accelerated phase (two patients), acute myeloid leukaemia (AML) (two patients), acute lymphoid leukaemia (ALL) (two patients), and refractory anaemia with excess blasts under transformation (tRAEB) (three patients). The patients received a median of 1.72 x 10(8) CD3+ cells/Kg (range: 0.58 x 10(8) CD3+ cells/Kg). Four patients were infused cryopreserved cells. Six patients received interferon alpha (IFN alpha) concomitantly. RESULTS: Seven patients (four CML, one AML, one ALL, one tRAEB) obtained complete remission (CR). Graft-versus-host disease (GVHD) was observed in all patients with CR and one without response. Marrow hypoplasia or severe bicytopenia occurred in four patients. Of all patients achieving CR, two died after relapsing within 3 months of DLI, while three others died of GVHD. Four patients had no response to DLI or were not evauable. Only two patients--both with CML--are alive 1096 and 374 days after DLI, the former in clinical, cytogenetic and molecular CR, and the latter in second CP after 2 months in CR. CONCLUSIONS: DLI results in CR in most patients with relapsing leukaemia or myelodysplasia after BMT, especially in CML patients. The anti-leukaemia effect is highly correlated with GVHD. This complication and marrow hypoplasia remain major causes of morbidity and mortality of this procedure. PMID- 10822761 TI - [MLL rearrangements in acute leukemias]. AB - PURPOSE: In present study we have studied MLL rearrangements in a serie of acute myeloid, lymphoblastic and biphenotypic leukaemias. PATIENTS AND METHODS: We analyzed a total of 11 cases: 9 acute myeloid leukaemias (M4 and M5 subtypes in FAB classification), 1 lymphoid leukaemia, and 1 acute biphenotypic leukaemia. We studied bone marrow samples from all patients by using conventional cytogenetic techniques and fluorescence in situ hybridization (FISH) with an MLL probe. We also analyzed the correlation between clinical features and genetic results. RESULTS: Cells from 6 patients showed to contain MLL rearrangements and these arose in all types of leukaemias included in this study. Some MLL rearrangements were detected by FISH in kariotypically normal cases or without cytogenetic evidence of 11q23 aberration. MLL gene duplication has been observed in two cases with M4 and biphenotypic leukaemia, respectively. The presence of MLL gene rearrangements does not shape a group of patients with a common clinical pattern. CONCLUSIONS: MLL rearrangements occurs in a wide variety of leukemias. These rearrangements should be screened by FISH techniques, taking into account that gene duplications could arise in cases with normal karyotype. MLL rearrangements appear to have a considerable clinicopathologic heterogeneity. PMID- 10822762 TI - [Major thromboembolic complications during oral anticoagulant therapy. Importance of level of anticoagulation]. AB - PURPOSE: The incidence of major thromboembolic complications in patients on oral anticoagulant therapy (OAT) and the correlation of this with the intensity of the OAT and the INR level at the time of the episode have been assessed in our study. PATIENTS AND METHODS: We have carried out a retrospective study including 1350 patients with an overall follow-up period of 6432 patient-years. The mean INR level throughout OAT and at the time of the mayor thromboembolic event were considered. The statistical analysis was performed by means of a survival analysis test. RESULTS: The incidence of major thromboembolic complications found in our study was 1.18/100 patient-years. Those patients with a mean INR below the therapeutic range showed significantly a higher risk (3.31 times higher) of suffering from some sort of major thromboembolic complication. Mean INR level at the time of the event was 1.9 and 47% of those patients had an INR level < 2 at the time of the thromboembolic complication. CONCLUSIONS: The probability of suffering a major thromboembolic complication for those subjects on OAT increases as the INR falls below the therapeutic range; therefore we must pay special attention to this factor in order to avoid any further recurrences. PMID- 10822763 TI - [Cryopreservation of hematopoietic progenitor cells]. PMID- 10822764 TI - [Transfusion related graft versus host disease. Indications for irradiated blood components]. PMID- 10822765 TI - [More reflections on our system of postgraduate teaching (II)]. PMID- 10822766 TI - [Acenocoumarol as the trigger of disseminated intravascular coagulation in a patient with cancer]. PMID- 10822767 TI - [Antithrombotic prophylaxis and central venous catheters in onco-hematological patients]. PMID- 10822768 TI - [Attenuated Fechtner syndrome: presentation of a new case with some variants]. PMID- 10822769 TI - [Clinical, biochemical and metabolic characteristics of a patient with a beta thalassemia/hemoglobin C genetic compound]. PMID- 10822770 TI - [Treatment with interferon alpha of the lymphoblastic transformation of chronic myeloid leukemia]. PMID- 10822771 TI - [ABL/BCR+ myelodysplastic syndrome. Report of a new case]. PMID- 10822772 TI - [Autoimmune thrombocytopenia in the clinical context of myelodysplasia with chromosome 7 deletion, intestinal tuberculosis, and chronic hepatitis caused by hepatitis C virus]. PMID- 10822773 TI - [Thyroid complications in Hodgkin's disease patients treated with radiotherapy and chemotherapy]. PMID- 10822774 TI - [Evaluation of an autotransfusion program in a county hospital]. PMID- 10822775 TI - Love and barriers to love. An analysis for psychotherapists and others. AB - It is vitally important that psychotherapists bring a strong understanding of the nature of love to their work with the many clients who are struggling, in one way or another, with love relationships. With this in mind, the present paper is designed to accomplish two purposes. The first of these is to provide an adequate answer to an old and perplexing question: "What is romantic love?," and to do so in a way that illuminates why this one relationship possesses the extraordinary importance and centrality in human existence that it so clearly does. The second is to identify and discuss the most common barriers to persons being able to love that are encountered in clinical practice. PMID- 10822776 TI - Thomas Mann's Tonio Kroger: a study of the protagonist's emergence from the schizoid to the depressive position. AB - The protagonist of Thomas Mann's novella Tonio Kroger is examined in terms of object-relational theory. This approach is briefly compared with the interpretation that might be offered by structural theorists. Kroger, the son of an authoritarian and prototypical Germanic father and an artistic and sensuous mother from the south, is tormented by what he sees as his dual heritage. Unable to accept that he can be both an artist and a respectable member of bourgeois society, he employs splitting as a primary defense mechanism both intrapsychically and interpersonally, projecting these opposing ideologies onto others including Hans, Inge, and Lisabetta. Alternately idealizing and demonizing these characters, projections of his internal conflict, he is unable to achieve an integrated sense of himself or others, functioning primarily in the schizoid position. His emergence from the schizoid into the more mature depressive position, having been foreshadowed in a series of dream sequences, occurs at the end of the work when he comes to accept the view that artistic impulses and bourgeois discipline are not incompatible. PMID- 10822777 TI - Intrapsychic momentum and the psychoanalytic process. AB - A clinically orientated definition of psychoanalysis is proposed and distinctions are made between interactive counseling, psychoanalytic psychotherapy, and psychoanalysis. Elements of the treatment, such as frequency, diagnosis, and use of the couch are found to be helpful tools rather than defining requirements. PMID- 10822778 TI - Praxis as a radical alternative to scientific frameworks for psychotherapy. AB - This article propounds that formal clinical frameworks do not provide answers to the significant technical problems that typically confront certain types of psychotherapists in their daily work. Specifically, it is claimed that for a certain class of therapies "normal science's" theoretical frameworks cannot logically entail (i.e., lead deductively to) therapeutic technique and clinical practices. Earlier publications explained this failure of formalized theories as stemming from structural features, i.e., from the impoverished representational capacity inherent in mathematization. Now a second kind of explanation focuses on functional limitations inherent in theories of the "behavioral objectives" type. Such models do not address the issue of values inherent in process and in goal selection, focusing instead on quantifying predetermined goals and measuring outcomes to assess whether the goals had been reached; process is reduced to an instrumental function. To counter both kinds of limitations--structural and functional--a radical and still largely unfamiliar framework, praxis, is proposed and introduced as an alternative to conventional theorizing. PMID- 10822779 TI - The couple assessment summary: a bridge from assessment to treatment. AB - Treatment of couples presenting for therapy typically necessitates a process of riding out the storm of contention, entering the contextual framework in which the couple is relating and developing an understanding of the couple's interactive as well as intrapsychic world. The therapist's emerging conceptualization of the relational problems addressed in the assessment process provides an important foundation for the treatment process to follow. This formulation is recorded in the "Couple Assessment Summary" (CAS), which is developed in the diagnostic stage of the treatment process. A clearly conceptualized and structured document, the CAS is the suggested vehicle for engaging the couple in the treatment process to follow. The CAS serves as a logical extension of the Couple Assessment Outline, which is a therapist's tool for engaging the partners in the process of self-assessment and formulation. The CAS offers the therapist's perspective on the problems presented to the couple for review. This paper will demonstrate the utilization of the CAS with couples in distress and will illustrate its use with a couple engaged in the assessment process. The CAS is seen as a useful method for developing a shared understanding of the problems presented as the assessment process is often fraught with tension, anger, and chaotic interaction. PMID- 10822780 TI - The use of the psychodynamic life narrative in crisis supervision. AB - The psychodynamic life narrative is a statement that is made to the patient that gives current emotional reaction meaning in the context of his life history, and shows it to be a logical and inevitable product of previous life experiences. Because the patient in crisis characteristically present feeling overwhelmed, confused and needy, there is a sense of urgency and helplessness which impacts on the resident therapist's intervention. Common countertransference reactions which can result are assuming omnipotent control, a grandiose rescue fantasy or passivity. The authors present a clinical vignette to illustrate the supervisory process and its vicissitudes in the use of the psychodynamic life narrative in supervision of crisis work. The early identification of the "rescuing" response led to the rapid formulation of the psychodynamic life narrative and a successful outcome. PMID- 10822781 TI - The Nazi's daughter: the therapist as Jewish mother. AB - The article gives an inside look at a case where posttraumatic stress is intertwined with disturbances in object relations and ego-identity. The patient was a victim of a brutal authoritarian father who had abused her sexually. The trauma left her powerless and isolated. The treatment was aimed at enabling the patient to achieve connection and empowerment. Hypnotherapy, psychoanalytic psychotherapy and hypnosis allowed the patient to understand and overcome symptoms and underlying conflicts in the context of the therapeutic relationship. In spite of regressions and resistances during the course of treatment, therapy yielded the following results: 1. Symptoms were in remission at the end of treatment, and at one- and two-year follow-ups. 2. During the course of psychotherapy the patient was able to process traumatic events cognitively and affectively. 3. There were corrective changes in relatedness modes. 4. Modifications in object internalization and representation were made. PMID- 10822782 TI - Psychotherapy of sexual dysfunction. PMID- 10822783 TI - The road to Rapallo. A psychiatric study. 1949. PMID- 10822784 TI - A bad case of mixed metaphors psychiatry, law, politics, society, and Ezra Pound. PMID- 10822785 TI - Group A streptococcus necrotizing fasciitis. AB - Necrotizing fasciitis due to Group A streptococcus has been observed with increasing frequency over the past decade. Appropriate management requires rapid recognition of this life-threatening infection and expeditious antimicrobial therapy as well as surgical debridement or excision of tissue. PMID- 10822786 TI - Cervical pain syndromes: primary care diagnosis and management. AB - This article discusses treatment of the most common musculoskeletal causes of cervical pain syndromes. A thorough history and physical exam can allow treatment of these disorders without use of unnecessary and costly testing in the current era of managed care. PMID- 10822787 TI - Diagnosis and treatment of osteomyelitis. AB - The diagnosis and therapy of osteomyelitis remains difficult despite recent advances. Clinical decision making is also difficult because of considerable variations in the types of disease observed and the lack of large comparative trials studying the variety of approaches. PMID- 10822788 TI - Newer indications for permanent pacemakers. AB - The role of pacemakers in cardiac disease continues to expand, and recent technology has allowed therapy to be individualized. Newer indications for pacemakers now include long QT syndrome, neurocardiogenic syncope, hypertrophic obstructive cardiomyopathy, and drug refractory atrial fibrillation. PMID- 10822789 TI - Prevention of NSAID-related ulcers. AB - The use of NSAIDs constitutes a significant risk for gastrointestinal bleeding and other ulcer complications. However, if they prove clinically effective in relieving arthritic symptoms, the new COX-2 selectively inhibiting NSAIDs may ultimately solve the problem of gastrointestinal toxicity with NSAIDs. PMID- 10822790 TI - Cardiac transplantation and other therapeutic options in the treatment of end stage heart disease. AB - This article focuses on the surgical treatment of heart failure. It reviews the advances and limitations of heart transplantation and gives a broad overview of emerging technologies in the treatment of end-stage heart disease. PMID- 10822791 TI - Hypokalemia and metabolic alkalosis: algorithms for combined clinical problem solving. AB - This article reviews an approach to patients with hypokalemia and metabolic alkalosis using the information obtained from spot urine chloride values, blood pressure determinations, and renin and aldosterone measurements in order to simplify clinical problem solving. PMID- 10822792 TI - Cyclosporine and graft coronary artery disease after heart transplantation. AB - Risk factors for graft coronary artery disease after heart transplant are discussed in relationship to cyclosporine dosages. Patients receiving a mean cyclosporine dose higher than 4 mg/kg/day had lower incidence of graft coronary disease than patients receiving lower dosages. PMID- 10822793 TI - The knee joint in rheumatoid arthritis. AB - Rheumatoid arthritis may present in the knee within a wide spectrum of manifestations. Disease progression, treatment protocols, surgical treatments and the possible complications of these treatments differ distinctly from those of osteoarthritis. PMID- 10822794 TI - Extracellular matrix effects on a neuroblastoma cell line. AB - Cells of various lines assume similar shapes when grown attached to substrates like coverslips. In contrast, cells cultured in a collagen and/or laminin matrix often assume a relatively normal morphology in comparison with their in situ counterparts. During investigations of neuroblastoma SH-SY5Y cells, an attempt was made to identify culture conditions which would cause the cells to assume a more regular shape. SH-SY5Y cells cultured on bare coverslips, on coverslips coated with rat-tail collagen, and in approximately 1 mm thick gels containing extracellular matrix components were compared. Striking differences were apparent when comparing the gel-cultured cells with cells cultured on coverslips. Cells grown in the gel formed ganglia-like clusters which generated bundles of neurites which targeted other 'ganglia'. The same cells grown on coverslips, whether or not they were collagen-coated, appeared unaware of the presence of other cells, and did not cluster, nor did they generate neurites. PMID- 10822795 TI - Accumulation of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in porcine preovulatory follicles after in vitro exposure to TCDD: effects on steroid secretion and cell proliferation. AB - The present experiments were conducted to test 2,3,7,8-tetrachlorodibenzo-p dioxin (TCDD) accumulation in the tissues, and its influence on cell proliferation and steroid secretion. A dose of 3.2 ng of TCDD/g tissue was added at the beginning of the culture, and the media were changed every 24 h or not changed till the end (96 h) of the culture. TCDD in the tissue was analysed by mass spectrometry, and the percentage of proliferating cells was measured using the MIB-1 labelling index. TCDD added to the culture medium accumulated in the tissues after 24 h (59.3%) and 96 h (81.2%) of exposure. The accumulative effect of TCDD was manifested by a reduction in the percentage of proliferating cells (53.5 and 33.8%, after 24 and 96 h exposure, respectively). A single exposure to TCDD had no effect on progesterone, reduced testosterone secretion and caused a significant increase in oestradiol secretion. Prolonged exposure to TCDD caused an increase in the concentration of the three steroids investigated in the culture medium. The results suggest that TCDD action is complex in the follicles. PMID- 10822796 TI - Methorchiasis in the residents of Novosibirsk area, Russia. AB - Infection with Methorchis bilis was recognized for the first time in the residents of Novosibirsk area (Russia). During a serological survey (37 patients in toto), it was possible to demonstrate that 48.5% of the serum samples tested possessed antibodies to Opisthorchis felineus antigens, 37.8% to both Opisthorchis felineus and Methorchis bilis antigens, and 13.5% to Methorchis bilis antigens only. PMID- 10822797 TI - The relationship between the Y chromosome size and the amount of autosomal Q heterochromatin in human populations. AB - The relationship between the basic quantitative characteristics of the Q heterochromatin (Q-HR) region variability of autosomes and of the Y chromosome in human populations was examined. A definite relationship between the mean number of Q-HR per individual, the distribution and frequencies of Q-HR on autosomes and the size of the Q-heterochromatin segment of the Y chromosome at the population level was shown to exist. The amount of autosomal Q-HR was lower in individuals with larger Q-heterochromatin segments on Y chromosomes, and vice versa. The hypothesis that the amount of chromosomal Q-HR in the genome of modern human populations may be under the control of natural selection, is discussed. PMID- 10822798 TI - Clastogenic effects of zinc chloride on human peripheral blood leucocytes in vitro. AB - The clastogenic effects of three different concentrations of zinc chloride on human peripheral blood leucocytes were studied in vitro. The highest concentration (1.5 x 10(-3) M) was lethal after 48 and 72 h of culture and no blast cells were formed. The two lower concentrations (3.0 x 10(-4) M and 3.0 x 10(-5) M) significantly reduced the frequency of cell division, induced chromatid breaks and damaged cells in frequencies significantly higher than in control experiments maintained in sodium chloride and in distilled water. PMID- 10822799 TI - Microalbuminaria and the risk of atherosclerosis. Clinical epidemiological and physiological investigations. AB - Microalbuminuria, i.e., slightly elevated albumin excretion in the urine, is considered a novel atheroslerotic risk factor. This supposition is based partly on two preliminary minor studies, partly on the current knowledge within the fields of diabetes mellitus and arterial hypertension. The aims of the present series of studies were 1) to examine whether a relationship exists between microalbuminuria and atherosclerotic cardiovascular disease in the general population, and 2) to illuminate possible pathophysiological mechanisms underlying this association. The studies were performed as sub-studies of the Copenhagen City Heart Study and the Monica Population Study. In the 3rd Copenhagen City Heart Study a cross-sectional analysis comprising 2,613 individuals without diabetes mellitus or renal- or urinary tract disease revealed a positive association between overnight urinary albumin excretion rate and a history of acute myocardial infarction. This association was independent of age, sex, conventional atherosclerotic risk factors, and glomerular filtration rate. Participants with a urinary albumin excretion rate exceeding the upper decile (7 micrograms/min) in the entire study population had a higher frequency of previous acute myocardial infarction than the others. In order to assess the time sequence of the observed association, this population will be followed prospectively. In the 1st Monica Population Study in Copenhagen County, 2,085 individuals without diabetes, mellitus, cardiovascular disease, or renal or urinary tract disease were followed for 10 years. Participants with a urinary albumin/creatinine concentration ratio exceeding the upper decile (0.65, mg/mmol) in the entire study population had a relative risk of 2.3 for developing ischaemic heart disease as compared to participants with a lower urinary albumin/creatinine concentration ratio. This predictive effect was independent of age, sex, and conventional atherosclerotic risk, factors, but it was not known whether participants with a urinary albumin/creatinine concentration ratio above the upper decile had severe subclinical atherosclerosis already at entrance to the study. A group of individuals with persistent microalbuminuria, and an age- and sex-matched control group with persistent normoalbuminuria underwent a clinical physiological and biochemical investigation program. None had developed clinically present atherosclerotic cardiovascular disease. Microalbuminuria was defined as a urinary albumin excretion rate exceeding the upper decile in the entire study population, normoalbuminuria as a lower urinary albumin excretion rate. Measurements of glomerular filtration rate and tubular function made it unlikely that the difference in urinary albumin excretion between the two study groups was due to local renal conditions exclusively, although reductions in both glomerular charge selectivity and size selectivity were observed in the microalbuminuric individuals. Individuals with persistent microalbuminuria had increased systemic transvascular albumin leakage to a level similar to that seen among individuals with severe clinical atherosclerosis. This could be explained neither by differences in blood pressure or concentration of plasma lipoproteins, both of which were more atherogenic in the microalbuminuric individuals, nor by differences in plasma volume or albumin concentration, antropometric factors, insulin sensitivity, or smoking habits. It is hypothesized that the systemic transvascular leakiness may also include lipoproteins, thus allowing for an increased lipid insudation into the vessel walls. The leakiness might be due to haemodynamic factors or structural or functional perturbations of the endothelium or the intracellular matrix beneath. Although endothelial dysfunction could not be demonstrated in the present studies, future research will focus on these possibilities. PMID- 10822800 TI - Autotransfusion of mediastinal blood after coronary artery bypass grafting. PMID- 10822801 TI - Cytokine actions on the thyroid gland. AB - Cytokines are integral components of the complex intercellular communication required to mount and control an immune response. The purpose of this review is to describe the influence of the most important cytokines on the thyroid gland in animal models and in humans and on isolated thyroid cells. We have used an in vitro system of monolayer cultures of human paraadenomatous thyroid cells for the study of the phenomenological actions of cytokines on the function of the thyrocytes. A biphasic, non-cytotoxic and reversible influence of IL-1 supporting a role of IL-1 in the physiological regulation of thyroid cell function was found. IL-1 in moderate to high concentrations and TNF and IFN-gamma all inhibited thyroid cell function. IL-1 induced release of NO and cGMP from the thyrocytes, but an inhibitor of nitric oxide synthase did not abolish the IL-1 induced inhibition of the release of Tg and cAMP from the TEC. The biochemical pathways by which IL-1 influences thyrocytes are not fully clarified. IL-1 beta inhibited the adenylate cyclase mediated pathways and stimulated the guanylate cyclase mediated pathways, and all the demonstrated IL-1 effects were counteracted by IL-1 ra indicating, that the effects were exerted through activation of specific IL-1 receptors on thyrocytes. The predominant effect of cytokines on the hypothalamic-pituitary-thyroid axis is inhibitory and the cytokines may play a role during physiological as well as pathophysiological conditions contributing to the euthyroid sick syndrome and AITD. A model for the pathogenesis of AITD is outlined. The trigger, of the autoimmune process in AITD is unknown. However, the earliest steps include the interaction between antigen presenting cells and Th cells. In the later phase antigen specific and non specific immune cells are recruited to the thyroid and an inflammatory infiltrate is built. During this process inflammatory mediators including cytokines, free nitric and oxygen radicals are released. A better understanding of pathogenetic mechanisms is crucial for an appropriate and effective management of AITD, and if possible, for its prevention. Further studies of the actions of these potent agents are one of the keys to a better understanding of the endocrine system both in health and in disease. PMID- 10822802 TI - Tobacco-related diseases: the role of gender. An epidemiologic study based on data from the Copenhagen Centre for Prospective Population Studies. PMID- 10822804 TI - [Genetic control of cytoplasmic male sterility in plants: state of the problem and current approaches]. AB - The organization and functions of genetic systems controlling cytoplasmic male sterility (CMS) in plants are reviewed. Mitochondrial genes that code for specific proteins disturbing the functions of mitochondria have been found by comparisons between CMS lines and fertile F1 hybrids and between forms originating from in vitro cultures (fertile revertants, cybrids). Nuclear fertility-restorer genes abrogate expression of these mitochondrial genes at the transcriptional or posttranslational level and thereby prevent synthesis of their protein products. A high mutability of the mitochondrial genome in cells cultured in vitro results in the elimination or reorganization of CMS-associated mitochondrial genes or in new mutations causing CMS. Several biotechnological methods are considered with respect to their applicability in identifying and transferring CMS-associated genes and in constructing new CMS forms. PMID- 10822803 TI - High human plasma levels of organochlorine compounds in Greenland. Regional differences and lifestyle effects. AB - INTRODUCTION: The purpose of the study has been to analyse data, collected for surveillance purposes under the Human Health Programme of AMAP (Arctic Monitoring and Assessment Programme, in Greenland, regarding geographical differences and lifestyle versus pollution load from persistent organic pollutants (POP), and to make possible exposure assessments. It comprises a regional study from six districts and an ongoing study of pregnant women and infants. METHODOLOGY: Biostatistical analyses of data collected in Greenland from 61 men and 10 women from six different districts (1997-98) and from 110 mother-infant pairs in the Disko Bay area (1996-97) and 223 mother-infant pairs (1994-96). The data consisted of questionnaire answers and bloodsamples from men, women, and newborn infants (cord blood) analysed for fatty acids, selenium and 26 POPs including 14 PCB-congeners and four toxaphenes. RESULTS: Strong regional differences were found, related to different intakes of marine food with very high PCB-loads among men from the East coast. The various POP-plasma levels were mutually correlated and strong correlations were found between POP-plasma concentrations in mothers and new-born, R > 0.9, p < 0.0001. Among the pregnant women 95% surpassed the Canadian concern level for PCB, and 60% of the men from Scoresbysund surpassed the action level. The association between reported monthly food frequency and POPs was relatively weak, but the POPs were strongly correlated with plasma and erythrocyte n-3/n-6 fatty acid ratios as biomarkers of marine food intake. Multiple regression analysis showed highly significant positive correlation between smoking, and POP-plasma levels, after correction for age, alcohol intake, marine food, plasma lipids and n-3/n-6 fatty acid ratios. CONCLUSION: As the most important determinants of high POP-plasma levels in Greenlanders we propose: age, high plasma n-3 fatty acids (marine food), East coast region, and being a smoker. PMID- 10822805 TI - [Variability of localization of retrotransposon copia ant its effect on adaptation in inbred strains of Drosophila melanogaster with the different rate of transposition]. AB - Three sublines of an inbred laboratory line of Drosophila melanogaster with the initial copia transposition rate 2 x 10(-2), 2 x 10(-3), and 5 x 10(-4) per copy per generation were reared for several dozen generations under conditions of low effective population size (by full-sib crosses or in a small mass culture of 10 females x 10 males). All six lines were tested for the transposition rate, location pattern, and copy number of copia in euchromatic genome regions and for fitness inferred from the intraspecific competition index. The copia transposition rate remained constant in both versions of the lines with an initially lower rate and decreased by an order of magnitude in both versions of the line with an initially higher rate. New copia insertions behaved as selectively neutral and were accumulated in the genome. Each new copy decreased fitness by less than 1% on average. Some of the existing unfixed insertions remained segregating after long-term inbreeding and were assumed to provide a selective advantage to heterozygotes. PMID- 10822806 TI - [Molecular genetic analysis of the Tn5041 transposition system]. AB - A study was made of the transposition of the mercury resistance transposon Tn5041 which, together with the closely related toluene degradation transposon Tn4651, forms a separate group in the Tn3 family. Transposition of Tn5041 was host dependent: the element transposed in its original host Pseudomonas sp. KHP41 but not in P. aeruginosa PAO-R and Escherichia coli K12. Transposition of Tn5041 in these strains proved to be complemented by the transposase gene (tnpA) of Tn4651. The gene region determining the host dependence of Tn5041 transposition was localized with the use of a series of hybrid (Tn5041 x Tn4651) tnpA genes. Its location in the 5'-terminal one-third of the transposase gene is consistent with the data that this region is involved in the formation of the transposition complex in transposons of the Tn3 family. As in other transposons of this family, transposition of Tn5041 occurred via cointegrate formation, suggesting its replicative mechanism. However, neither of the putative resolution proteins encoded by Tn5041 resolved the cointegrates formed during transposition or an artificial cointegrate in E. coli K12. Similar data were obtained with the mercury resistance transposons isolated from environmental Pseudomonas strains and closely related to Tn5041 (Tn5041 subgroup). PMID- 10822807 TI - [The phenomenon of predetermination of the cytoplasm upon interaction between alleles of the ADE2 and ADE13 in the yeast Saccharomyces cerevisiae]. AB - Our previous data showed that mutation ade13-1, blocking steps 8 and 12 of purine biosynthesis in the yeast Saccharomyces cerevisiae, caused the inability of strains manifesting this activity to grow on the complete nutrient medium with glucose in addition to the loss of adenylosuccinate lyase activity. It was also determined that the ade2-D mutation, inactivating aminoimidasole ribonucleotide carboxylase (the enzyme of step 6), suppressed this phenotypic manifestation of ade13-1; i.e., the ade2-D mutation restores the ability to grow on this medium. When spores of a hybrid that contained both mutations in the heterozygote were germinated on the YEPD medium, almost complete viability of segregants with genotypes ADE2 ADE13 and ade2-D ADE13 and the absence of ADE2 ade13-1 growth were observed. The number of growing segregants ade2-D ade13-1 amounted to approximately half of the possible number. In this work, a decrease in the proportion of segregants with this genotype was shown to occur only when they were obtained as a result of the segregation of hybrids with the normal allele (ADE2) in the heterozygote. The proportion of segregants with genotype ade2-D ade13-1 did not decrease upon segregation of hybrids similar in the genetic background and containing the ade2-D mutation in the homozygote and ade13-1 in the heterozygote. Spores with this genotype formed in the diheterozygous diploid were able to germinate on a medium containing glycerol and to further grow on a medium with glucose. The data suggest that, when a product of the normal ADE2 allele or of another gene, the synthesis of which is stimulated in the presence of this allele, enters spores with genotype ade2-D ade13-1 during meiosis, some of these spores lose their ability to grow on the medium with glucose; i.e., the ADE2 allele can be phenotypically expressed in the spores that did not contain this allele. This phenomenon is similar to the maternal effect known in some species of animals from various systematic groups. PMID- 10822808 TI - [Wolbachia--a new bacteria causing sex ratio bias in the two-spot lady-bird Adalia bipunctata L]. AB - Some of the male-killing lines of the two-spot ladybird Adalia bipunctata L. isolated from the populations of Moscow and Tomsk and having a female-biased sex ratio were found to be infected with a bacterium of the genus Wolbachia. This fact is the first demonstration of the ability of Wolbachia to kill males of a host insect. The coexistence of females infected with different male-killing bacteria was recorded in the population of Moscow. PMID- 10822809 TI - [The level of dysgenic sterility and recessive mutations induced in laboratory strains of Drosophila melanogaster exposed to chronic low-dose gamma irradiation]. AB - Recent investigations showed that genetic instability accounts for many radiobiological effects. However, mechanisms underlying this phenomenon are still poorly understood. Assuming that mobile genetic elements may be involved in the induction of genetic instability, we studied parameters that characterize the activity of these elements in Drosophila melanogaster: hybrid dysgenesis and the level of recessive lethal mutations. In our experiments, we used D. melanogaster strains that differed in the type of hybrid dysgenesis (P-M and H-E). It was demonstrated that chronic exposure to radiation leads to substantial changes in the genetic structure of a population and an enhanced level of dysgenic sterility. Our results indicate that genetic instability and adaptation to the effect of chronic gamma-radiation are associated with the radiation-induced mobilization of mobile genetic elements. PMID- 10822810 TI - [Heavy heat shock induces genetic variation in a polygenic system of a quantitative trait in Drosophila]. AB - Results of two experiments dealing with positive and negative selection on the quantitative trait radius incompletus in an isogenic line of Drosophila melanogaster after heavy heat shock (HHS) are presented. Selection was not effective in the control without HHS. In experiment 1, in which offspring of HHS exposed males lacked transposition induction, selection also was ineffective. By contrast, selection was highly effective in offspring of males that responded to HHS exposure by transposition induction. Thus, HHS, which is not mutagenic, generates genetic variation in a polygenic system of a quantitative trait via transpositions and excisions of mobile genetic elements. In experiment 2, positive and negative selection was conducted in three replicates, which showed concerted dynamics of the selected trait. This means that the trait dynamics is mainly related to the nearly deterministic process of accumulation of active polygenic alleles rather than to genetic drift. The induced variation of polygenic systems promotes rapid selection of "champion" genotypes. This variation is probably associated with "soft" modification of polygene expression by adjacent MGE copies. PMID- 10822811 TI - [Mating success and courtship ritual in strains of Drosophila melanogaster carrying mutation flamenco]. AB - Mating success was examined in groups of Drosophila melanogaster carrying mutation flamMS (SS, MSn1-2, and MSn1-3) and in wild-type flies. The proportion of normally copulating males was significantly lower in the mutant strains. The reduction in mating efficiency was caused by changes in male behavior rather than in female attractiveness. Individual analyses showed that male mating behavior in strains carrying flamMS was qualitatively and quantitatively different from that in the wild-type strain Canton S. The proportion of males that performed consecutive courtship stages was significantly lower in the mutant strains. The sequence and duration of some courtship stages (in particular, orientation and wing vibration) in mutant flies was shown to be altered. The significance of the flamenco locus in regulation of processes occurring at the organismal level are discussed. PMID- 10822812 TI - [The effect of genotype on the rate of regeneration of plants in a microspore culture of Triticum aestivum L]. AB - The effect of genotype on the productivity of in vitro microspore culture of wheat (Triticum aestivum) was studied. Wheat cultivars and hybrids bred in Kazakhstan and used in the programs on breeding new high-productivity and disease resistant cultivars and forms of plants were tested. The genotypes that are responsive to androgenesis have been found among the cultivars tested: cultivars Kazakhstanskaya 4, Kazakhstanskaya 5, and hybrids obtained from crosses with the participation of these cultivars (K-6, B-5, and B-10). Such genotypes are recommended to be used as model objects in the studies on androgenesis and plant biotechnology. Based on the use of haploid technology, wheat dihaploid lines were created that are currently tested by breeders. PMID- 10822813 TI - [Identification of Sorghum genes responsible for resistance to Green bug]. AB - Genes responsible for resistance to greenbug (Schizaphis graminum Rond.) were identified in sorghum. The dominant (Sgr1) and recessive (Sgr2) genes for resistance were revealed in sample k-457 (PI264453, United States). The samples i 589430 (PI264453, Spain) and k-3852 (Sarvasi, Hungary) carry gene Sgr1. These accessions are assumed to also have gene Sgr2. The samples k-9921 (Shallu, United States) and k-9922 (KS-30, United States) have incompletely dominant resistance gene Sgr3. A symbol Sgr4 was assigned to the dominant gene from sample k-6694 (Deer, United States). The dominant Sgr5 and recessive Sgr6 genes were revealed in the samples k-1362 (Durra Belaya, Syria) and k-1240 (Dzhugara Belaya, China). The cultivar Sorgogradskoe (k-9436, Rostovskaya oblast) has gene Sgr5. The samples k-10092 (Odesskii 360, Ukraine) and k-5091 (Cherhata, Marocco) are assumed to have genes Sgr5 and Sgr6. Sample k-924 (Dzhugara Belaya, China) is protected by the dominant gene Srg7 and recessive gene Sgr8. Sample k-923 (Dzhugara Belaya, China) has at least one of these genes. Two dominant complementary genes for resistance (Sgr9 and Sgr10) were revealed in sample k-930 (Dzhugara Belaya, China). One of two dominant genes of sample k-1237 (Dzhugara Belaya, China) was assigned the symbol Sgr11. Genes Sgr5-Sgr11 responsible for resistance to greenbug are new and were not previously used in breeding. PMID- 10822814 TI - [Effect of genetic diversity in Hordeum vulgare L. and Secale cereale L. on their crossability and the development and viability of hybrid embryos and plants]. AB - A study was made of the effect of genetic diversity in cultivated barley and rye on seed setting, embryonic development, and viability of barley x rye hybrids grown in vitro and planted. The variation in seed-setting frequency was shown to be determined mainly by vegetation conditions and random factors, while the variation in proportion of seeds with embryos among the total seed set was determined by the genetic diversity of barley and rye. Selection of barley genotypes was shown to increase the yield of viable barley x rye hybrids. Based on the phenotypic analysis of barely x rye hybrids obtained in various combinations, low viability and hybrid lethality were attributed to the hybrid necrosis genes expressed from early ontogeny. These genes are assumed to be widespread among barley and rye varieties. PMID- 10822815 TI - [Dynamics and genetic determination of transformation of alpha macroglobulins allogroups in different populations of domestic pigs]. AB - The results of genetic studies of transformation of alpha-macroglobullin (AM) allogroups (i.e., fixed allotype combinations) in pigs are presented. Population and hybridization studies showed that the phenomenon discovered was based on activation/inactivation of the AMI allotype expression, which depended on the genetic environment and interlocus relationships. Genetic and selection bases, as well as the significance of the transformation phenomenon described, are discussed. PMID- 10822816 TI - [Detection of differences in liver protein composition between recombinant strains of mice with different ThioTEPA sensitivities]. AB - The sensitivity of recombinant mouse strains subjected to 23-27 generations of inbreeding to the clastogenic effect of thioTEPA (triethylene thiophosphoramide) was reestimated, and their characteristics were confirmed. Six 1XC3 recombinant strains were obtained from crossing strains 101/H x C3H/Sn, which differed from one another with respect to the sensitivity to thioTEPA. The protein composition of the liver tissue was studied in the recombinant strains by means of two dimensional electrophoresis. Interstrain differences with respect to five liver proteins were found, which were correlated with the differences in the response to the mutagen. PMID- 10822817 TI - [Phenotypic and genotypic characteristics of russian hairless cats]. AB - A novel mutation that causes the loss of hair was found in Russian cats. In contrast to hairless cats known in other countries (Sphinx cats of Canada, Great Britain, France, and Germany, etc.), in which the loss of hair is inherited as a monogenic recessive trait, in Russian hairless cats this trait is determined by a semidominant gene with the participation of other genes. PMID- 10822818 TI - [Dynamic of marriage structure in three cities of Ukraine from 1960 to 1992]. AB - Marriage structure was studied in the city of Kiev and in two cities of the Sumy oblast, Shostka and Trostyanets. Ethnic affiliations and birthplaces of persons contracting marriage were analyzed as the main characteristics of population genetic diversity. The ethnic composition of persons contracting marriage and the proportions of mono- and interethnic marriages remained almost unchanged during one generation. The majority of the persons contracting marriage were Ukrainians (66-91%); among other ethnic groups, only Russians considerably contributed to ethnic diversity (up to 26%). During the period studied, coefficients of marital migration substantially decreased in Kiev (from 0.66-0.82 to 0.34) and Shostka (from 0.72 to 0.52) and changed only insignificantly in Trostyanets. Outbreeding was estimated based on the migration parameters, exogamy level, and marital migration distances. The outbreeding level in the Shostka population (100,000 people) was comparable with that for the considerably larger Kiev population (two million people); however, it was significantly higher than that for the Trostyanets population, the size of which was close to the size of the Shostka population. It is supposed that "migration stress" may unfavorably affect the adaptive genetic structure of the Shostka population. PMID- 10822819 TI - [Parallel variation in mitochondrial DNA type 16278T in genome of the migrant population of Northeast Asia and other caucasoid populations]. AB - Types of mtDNA with +16274EcoRV, which had been previously found in East Slavonic residents of Magadan, were identified on the basis of published data on nucleotide sequences of the hypervariable segment I (HVSI) of the mtDNA control region. These mtDNA types occurred in Slavs at a frequency of 7-14% and were assigned to four mitotype groups (1, 2A, 3B, and 5) restricted to Caucasoid populations. Regarding the evolution of mtDNA, this fact indicates that the C-T transition at position 16,278 arose independently in various groups of mtDNA types in Caucasoids. A new subgroup was isolated in Caucasoid group 5. One of the key mutations in this subgroup was variant +16398HaeIII (transition A-G at position 16,399), which was earlier detected in Russian populations at a relatively high frequency. PMID- 10822820 TI - [Analysis of the association between the T113M polymorphism of the human il-9 gene and bronchial asthma]. AB - The T113M polymorphism resulting from the missense mutation in exon 5 of the human interleukin 9 (IL9) gene was tested for association with bronchial asthma (BA). The genotype frequency analysis did not reveal a deviation from the Hardy Weinberg equilibrium. A comparison of the genotype frequency distributions in a control group of healthy individuals and in patients with BA suggested an association between T113M and the clinical phenotype. However, this association was not confirmed by the affected family-based association control (AFBAC) or the transmission/disequilibrium test (TDT). PMID- 10822821 TI - [Polymorphism of immunological and biochemical marker genes in rural populations of the Tuva Republic]. AB - This article continues the series of publications on the population genetic structure of the Tuva Republic. The polymorphism of immunological (ABO, MN, and the Dd locus of Rhesus) and biochemical (TF, GC, HP, PGD, PGM1, ACP1, and ESD) marker systems was studied in three rural populations of the Tuva Republic: the Shinaanskii, Todzhinskii, and Bai-Taiginskii populations (the Kungurtug, Toora Khem, and Teeli villages, respectively). Genetic subdivision of the populations and genetic distances between the Tuvinian populations and the populations of neighboring regions were estimated. Tuvinians were demonstrated to be genetically heterogeneous. Data on their population-genetic structure with respect to several classical marker systems agree with the results obtained for quasigenetic (family names) and molecular (mtDNA) markers. Prolonged isolation of individual populations in the republic promoted formation of specific patterns of gene frequencies in some of them. These patterns account for differences between Tuvinians and other populations belonging to the Altaic language family. Tuvinians in general are genetically closer to Mongolian populations inhabiting the regions bordering the Tuva Republic than to southern Altaians. PMID- 10822822 TI - [Computer analysis of band variability in DNA fingerprints]. AB - Through the example of the distribution of PCR products DNA matrices of mouse tail tissue, a method of comparative analysis of DNA fingerprints is described. The PCR products were obtained using a 20-mer random primer flanking the Atp1b2 locus on mouse chromosome 11. A software program was designed that permits the simplification of comparison of DNA fragments variability or polymorphism detected on electrophoregrams from different individuals. PMID- 10822823 TI - [A study of polymorphism of the D2 dopamine receptor gene in the population of the Volga-Ural Region]. AB - Using polymerase chain reaction (PCR), TaqI polymorphism for the D2 dopamine receptor gene (DRD2) was studied in eight populations of the Volga-Ural region that belong to the Turkic (Bashkirs, Tatars, and Chuvashes), Finno-Ugric (Maris, Komis, Mordvinians, and Udmurts), and Eastern-Slavonic (Russians) ethnic groups. Significant differences in the distribution of genotype frequencies were found between the Tatar population belonging to the Turkic branch of the Altaic linguistic family and the Mari and Mordvinian populations belonging to the Finno Ugric branch of the Ural family and between the Tatar and Bashkir populations belonging to the Turkic ethnic group. PMID- 10822824 TI - [Effect of chronic low-dose irradiation and etoposide on the life spain of Drosophila melanogaster strain mei-41]. AB - Longevity is a temporal characteristic dependent on the level of equilibrium between the damaging and restorative processes in an organism. This is a complex parameter determined by both genotypic and external factors. In experiments with the mutant strain of Drosophila melanogaster mei-41D5, it was demonstrated that chronic exposure to low-dose radiation can change the longevity of flies. A decrease in the longevity of both males and homo- and heterozygous females of this strain was also caused by specific inducers of apoptosis. We suggest that apoptosis plays a certain role in the aging of an organism and that the dominant gene mei-41D5 takes part in determining longevity in Drosophila. PMID- 10822825 TI - [The study of interaction between paralogous tandem repeats stellate and suppressor of stellate in the genome of Drosophila melanogaster]. AB - Testis-specific expression of tandemly repeated Stellate genes, located in eu- and heterochromatin regions of the X chromosome of Drosophila melanogaster, is suppressed by homologous Suppressor of Stellate repeats located on the Y chromosome. Using transgenic lines, we have demonstrated that three Su(Ste) copies failed to change the expression of the reporter construction carrying the bacterial beta-galactosidase gene under control of the Stellate gene regulatory sequence. Possible mechanisms of the Su(Ste) repeat suppressor activity are discussed. PMID- 10822826 TI - [The molecular conception of meiosis passes the test. The review of the Fourth European Conference on meiosis]. PMID- 10822827 TI - [Review of the production of ostrich meat]. AB - Ostrich meat was originally considered to be a by-product of the leather industry. Ostrich farming focused on the production of good quality hides and was mainly based on experience. Since a few years there has been a move from hide production to meat production; however, little is known about ostrich meat production which has consequences for legislation. There is insufficient knowledge about the factors influencing muscle growth. The strong growth of ostrich meat production in the EU member states should be discussed because the climatic conditions are not optimal and the price of the meat may remain high compared to meat of broilers, pigs, and cattle. Information programmes have to be set up for all members in the production chain. The present method for stunning of ostriches has to be changed in most slaughterhouses. In this study the effects of electrical and mechanical stunning on unconsciousness, duration of unconsciousness, behaviour, and meat quality parameters were examined. At least 500 mA is needed to stun ostriches effectively, and they can be killed using a short-stick interval or a long stunning duration. A modified captive needle pistol, using air pressure, is an alternative to electrical head-only stunning. PMID- 10822828 TI - [Is the carrying out of educational fetotomy in the living animal accounted for ethically?]. PMID- 10822829 TI - [Until death follows? Yes, of course!]. PMID- 10822830 TI - [Appropriate care for large grazers in nature]. PMID- 10822831 TI - [Genetic knockout--the first steps and outlook for a neurophysiology of behavior]. AB - A review of modern data on genetic knockout strategy application to study the brain neurotransmitters and their role in the regulation of behavior. Advantages and shortcomings of genetic knockout of receptors and the enzymes of neurotransmitters metabolism models in comparison to other methods are discussed. Data on the effect of genetic knockout of various types of opioid, dopamine, serotonin and adrenoreceptors as well as enzymes in biosynthesis of catecholamines and serotonin on physiology and behavior is adduced. The data provide evidence that genetic knockout reproduces a principal effects of the lack of receptors and enzymes and allows to find new yet unknown properties. Mouse strains with genetic knockout represent unique models of hereditary neuropathology. At the same time the data presented clearly demonstrated that the lack of a single type of receptor or enzyme does not lead usually to disorganization of regulated by them physiological functions and behavior. The data witness to the complexity and multifactoriality of their regulations and evidenced the great compensatory potentials of organism. PMID- 10822832 TI - [Macrophagal polykaryons]. AB - The mechanisms of formation, morphology and functions of macrophagal polykaryons are discussed. These giant multinuclear cells are formed by means of macrophagal cell fusion ("natural hybridization") and specialized on the extra- and intracellular processes of resorption of the foreign body and tissues of the proper organism. The formation of macrophagal polykaryons seems to be one of the possible manifestation of reactive histiocytosis in chronic inflammation and tumor growth. The role of macrophagal polykaryons in such disease as giant cell arteritis, giant cell granuloma and fibrohistiocytic tumors is considered in details. It is proposed that one and the same mechanism of regulations served as a basis of developing of both osteoclastic remodelling of the bone tissue (morphogenetic process) and resorption of the foreign body and tissues of the proper organism in inflammation in the process of evolution. PMID- 10822833 TI - [Beta-adrenergic receptors of the normal heart and in heart failure]. AB - The heart is often refereed to as an "beta-adrenergic organ" because beta adrenergic agonists are powerful stimulants of cardiac contractility. Catecholamines acting through beta-adrenoceptors produce both positive inotropic and chronotropic effects in human heart. It is now generally accepted that in human heart both beta 1- and beta 2-adrenoceptors coexist. beta-Adrenergic transduction system consist of membrane-bound beta-receptors, the effector enzyme adenylyl cyclase and guanine nucleotide-binding transduction (G) proteins. Repeated long-lasting agonist stimulus evokes homologous or heterologous desensitization of transduction system. Chronic heart failure accompanies with decreased responsiveness to beta-adrenoceptor agonists and is thought to exacerbate the loss of cardiac contractility. Depending on the etiology of heart failure abnormalities of the beta-receptor-G protein-adenylyl cyclase system result from a reduced of beta 1-receptors, uncoupling of beta 1- or beta 2 receptors, alteration of G-protein function, or decreased catalytic subunit activity of adenylyl cyclase and enhanced expression of beta-adrenoceptor kinase. The model most widely used is that of circulating lymphocytes that contain a homogeneous population of beta 2-adrenoceptors. The biochemical and pharmacological properties of human lymphocyte beta 2-adrenoceptors are quite comparable to those of heart beta 2-receptors. The analysis of lymphocyte beta 2 adrenoceptor-adenylyl cyclase system can be used as a model for long-term regulation of human cardiac beta 1- and beta 2-adrenoceptors only if serial changes in response to administration of non-selective beta-adrenergic agonists or antagonists are being investigated. This review concentrates on beta adrenoceptors in human healthy heart and in heart failure and also on lymphocyte beta 2-adrenoceptors and on the changes of these receptors properties under the influence of some cardiotropic drugs. PMID- 10822834 TI - [Mechanoelectrical feedback in the healthy heart and in the heart with pathologies]. AB - The article discusses the issues of possible connection between mechanical phenomena in myocardium and the electrical processes. Not only cardiomyocytes, but also cardiac fibroplastic are considered as substratum for the mechanisms of mechano-electrical feedbacks. Cardiomyocytes and fibroplastic of healthy animals demonstrate the mechano-electrical feedbacks, which essentially mean that stretching of the cardiac tissue within the physiological limits to 2 mN changes the electrophysiological cell processes. Close to 90% repolarization potential of cardiomyocytes action the mechano-induced depolarization develops; over the background of depolarization, when it reaches the threshold values, extra potentials of action are generated. In fibroplastic, membrane mechano-induced hyperpolarization develops; as result of cellular interaction it may develop hyperpolarization of pacemaking cells of the right auricle and slow the cardiac rythm down. In case of a pathology, for instance, infarct of the left heart ventricle modification of electric cell activity was detected at quite low values of tissue stretching up to 0.2. mN. Mechano-induced depolarization of cardiomyocytes of animals affected by infarct develops at 50% level of repolarization phase of action potential, or at 90% of repolarization phase. In the former case development of mechano-induced depolarization coincides with the period of absolute cell refractering. Extra action potential develops immediately after the refractering phase when the mechano-induced depolarization shifts the membrane potential towards threshold values. In the latter case the mechano induced depolarization transforms into extra action potential. With further stretching fibrillation develops. In fibroplastic the values of mechano-induced membrane hyperpolarization grow with greater scope of infarct damage. Magnitude of mechano-induced hyperpolarization of auricle fibroplastic taken from the animals with infarcts shows dependence on the period of remodelling if stretching is tissue is a standard parameter. With prolongation of the remodelling period the value of mechano-induced fibroplastic hyperpolarization diminishes. The problem of developing the combinations eliminating mechano-induced cardiac arrhythmia is raised. PMID- 10822835 TI - [The methodology of the theory of self-organization in the development of concepts on the physiological mechanisms of vestibular reactions]. AB - Sources of modern vestibulology's crisis status are determined. New methodological questions of object exploration of vestibular function and vestibular reactions are marked. New theoretical views are developed, some results of these theoretical views's practical realization are presented, perspective possibilities of new approach are defined. PMID- 10822836 TI - [An historical-physiological analysis of the discussion between I. P. Pavlov and V. M. Bekhterev on the problems of localizing functions in the cerebral cortex]. AB - This article is the first of the domestic publications on physiology and medical history, which highlights the discussion between I. P. Pavlov and V. M. Bekhterev on the issue of localization of functions in human cerebral cortex: it provides information of the visit by I. P. Pavlov to the clinic of V. M. Bekhterev; it discusses the role of V. M. Bekhrerev's students who described the cortex zones of tonotopics, gustation, regulation of salivation and stomach secretion, which I. P. Pavlov denied. Unlike articles on factography and history of physiology, which in various ways praise the scientists, this article is based of the modern approaches of medical history and scientific knowledge, in particular, it provides a retrospective of the major facts of the discussion. PMID- 10822837 TI - [The mechanisms of the formation and the role of the oscillatory activity of the neuronal populations in brain systemic activities]. AB - Experimental findings and theoretical concepts that have led to a new insight into the neurophysiological mechanisms underlying information processing and brain state are presented in this review. It is assumed that the brain information processing associated with elementary neuronal assembles is reflected by oscillations in the range of 30-70 Hz and their spatio-temporal organization in millisecond intervals. He functional brain state associated with the non specific systems is reflected in synchronization or desynchronization of activity of large neuronal populations in the frequency range of 20 Hz. PMID- 10822838 TI - [A model of color vision in trichromats and protans]. AB - A model which explains the human vision protanopic deficiency and its biologic prototype with the absence of red-absorbing pigment (rabbit) was constructed from neuron-like elements. In behavioral experiments and by means of evoked potential technique it was shown that the rabbit's color space is characterized by a spherical four-dimensional with a reduction of red-coding area. Similar spherical four-dimensional structure of color space is characteristic for a group of protanopic human subjects. The perceptive space of another group of protanopic subjects (protanomals) is characterized by a reduction of both parts of the red green opponent axis. These disorders are reproduced in the model either by a loss of some color-coding elements (the absence of the red-absorbing pigment as in protanops) or a shift of the spectral characteristics of the red pigment towards those of the green one (protanomals). PMID- 10822839 TI - [The voluntary regulation of consciousness levels]. AB - A capability for voluntary regulation of the intensity of spontaneous mental verbal associations (without external feedback) was found in several persons from 28 examined subjects. Due to this capability, these persons could control the level of consciousness and voluntary produce the corresponding EEG changes from the EEG enhancement to substantial decrease in its power (to appearance of the flattened EEG). PMID- 10822840 TI - [The electrodermal indices of the subjective perception of performance errors during drowsy changes in consciousness]. AB - The study was aimed at searching for objective criteria of subject's perception of errors in performance due to drowsiness and estimating the time between the onset of errors and the moment of their awareness. Healthy subjects (n = 64) with the pronounced EEG alpha were examined under conditions without sleep deprivation. Experiments (n = 280) lasted for 40 min. During the experiments, the EEG, EOG, EDR, EKG, and button pressings were recorded and subject's reports were registered. The subjects were waken up after the onset of errors or 20 min after the beginning of the performance independently of errors. The error onset was shown to be preceded by the EEG "drowsy pattern" and decrease in the rate of spontaneous EDR. The performance reappearance after the error was accomplished by alpha-rhythm independently from error awareness. The interrogation of subjects after the error and activity revival demonstrated a correlation between the error subjective perception and appearance of the EDR. The interval between the error and the first following EDR (mean 10.1 s) was significantly (p < 0.001, t-test for dependent samples) shorter than the interval between the error and the last preceding EDR (mean 69.8 s). It is suggested that the subjective perception of an error is a significant endogenous stimulus, which evokes the orienting response and accompanying sympathetic activation. PMID- 10822841 TI - [The mosaic structure of the high-frequency range of neocortical electrical activity in dogs]. AB - The study was aimed to reveal the subbands of correlated changes in power spectral density of brain electrical activity (EA), including the low-voltage (up to 10 microV) high-frequency (HF) components (40-200 Hz) in dogs during instrumental conditioning by means of factor analysis. The values of the EA spectral density in interstimulus intervals calculated with a resolution of 1 Hz were used for subsequent factor analysis (the standard principle component technique with varimax rotation). Twenty factors could explain about 80% of total variance. The groups of frequencies which were presented by comparatively narrow peaks (2-3 points) of high loads (more than 0.6) of single factors ("stable" factors) were taken into consideration. In the process of conditioning the factor organization of the EA became substantially complicated, the number of the "stable" factors increased. It was originally shown that the high loads of these factors divided the HF band in comparatively narrow frequency subbands, which appear to reflect the functional mosaics in the neocortex. PMID- 10822842 TI - [The state-dependent action of neuropeptides and monoamines on the background neuronal activity of the medial septal area in hibernating animals]. AB - Neuronal activity of the medial septal area (MS-DB) was recorded extracellularly in brain slices from two groups of Yakutian ground squirrels Citellus undulatus- hibernating (winter period) and actively waking (summer period). Effects of three neuropeptides identified in the brain of hibernators (TSKYR, TSKY, and DY) and of two monoamines (serotonin and noradrenaline) on spontaneous activity were analyzed. All neuropeptides reversibly changed the levels of the background activity, but in the hibernating ground squirrels (HGS) the level of reactivity (47-56%) was significantly higher than in the waking ground squirrels (WGS, 25 30%). Serotonin also showed some tendency to higher efficacy in the HGS. Only noradrenaline was equally effective and had absolutely dominating excitatory effect in both states, although the level of excitation in the HGS was higher. All other substances evoked excitatory and inhibitory effects in various proportions. Their distribution was state-dependent, the rate of development, intensity and duration of the effects were greater in the HGS. The experiments confirmed the data on higher excitability and reactivity of the septal neurons in the state of hibernation. It is suggested that the tested neuropeptides may participate in the control of hibernation. PMID- 10822843 TI - [The differences between sexes and strains in the capacity to acquire a passive avoidance conditioned reflex in KLA- and KHA-strain rats]. AB - The interstrain differences in passive avoidance conditioning were studied in male and female KHA (Koltushi High Avoidance) and KLA (Koltushi Low Avoidance) rats. These strains were selected for the rate of acquisition of active avoidance in a shuttle box. It was shown that the passive avoidance was substantially better acquired in the KLA strain than in the KHA. In females KHA rats the capability for passive avoidance conditioning depended on the estrus phase: the conditioning was impossible in proestrus. PMID- 10822844 TI - [Is "scopolamine-induced amnesia" in rats the result of state-dependent learning?]. AB - The effects of a single and repetitive administration of m-cholinoblocker scopolamine (Sc) to male rats on retention of step-through passive avoidance (PA) or active avoidance (AA) in a shuttle-box were compared. In case of PA Sc (1 mg/kg) was injected i.p. only 30 min before training, only 30 min before testing, or both before training and before testing. In case of AA Sc (0.5 mg/kg/day) was injected i.p. only 15 min before each training session or both before training and before testing (44 days after achievement of learning criterion). The PA and AA retention were impaired only in the experiments, where the drug was administered before training, but did not differ from control, when Sc was injected twice. The Sc-induced amnesia (like many other cases of memory deficits) is suggested to be a manifestation of state-dependent learning. Similarity between the brain state during memory consolidation and during the retention test is necessary for recollection. PMID- 10822845 TI - [Oscillations in the oxidation-reduction potential of the brain tissue in rats developing during wakefulness and slow-wave sleep]. AB - Variations of the brain cortex redox state potential (E) were recorded in freely moving white rats (mass of 300-350 g) with implanted platinum electrodes (with the platinum reference electrode in the nasal bone) during sleep-wake cycles. It was found that transitions from the slow-wave sleep to wakefulness were accompanied in the number of cortical areas (metabolic-active sites) by the E rise, while the transitions from the wakefulness to slow-ware sleep were associated with a drop of E. However, the episodes of the short-term arousals during the slow-wave sleep were accompanied by the respective decreases in E thus forming the irregular E variations (1.5-3 min in duration). It was also found that the oscillations of a typical pattern (quasisinusoidal with the frequency of 10-20 osc/min and the amplitudes up to several mV) could take place in the metabolic-active cortical sites. These oscillations were defined as fast E oscillations. During the slow-wave sleep, the less regular oscillations with the lower frequency (1.2-10 osc/min) and higher amplitude were recorded in the same cortical sites. These oscillations were defined as slow. It is suggested that the fast metabolic oscillations of wakefulness are mainly controlled by the mitochondria of neuronal populations, whereas the slow metabolic oscillations which occur in the slow-wave sleep are related with glycolysis in populations of glial cells. PMID- 10822846 TI - [The correction of the manifestations of a neurosis-like state in white rats by using the vitamin complex Aekol]. AB - Behavior of neurotized white rats was studied in two experimental situations: during training for passive and active avoidance of electroshock applied to the limbs and during testing the acquisition of the conditioned avoidance. It was shown that in behavior of rats in the neurosis-like state the signs of anxiety and fear dominated, the orienting-exploratory behavior was suppressed. The state correction was performed by per os introduction of the natural vitamin complex Aekol during the neurotization of animals. PMID- 10822847 TI - [The effect of "social stress" during rat pregnancy on the anxiety level of the progeny]. AB - Anxiety and locomotion were studied in offsprings of female rats subjected to everyday stress (one a day being displaced into another cage with pregnant rats) during the 3d stage of pregnancy. At the age of 1 month, the prenatally stressed rats had higher anxiety and lower locomotion in comparison with control animals. At the age of 3 month, the prenatally stressed females did not significantly differ from the control in the level of anxiety and locomotion, while the males demonstrated lower ambulation than the control animals. PMID- 10822848 TI - [The effect of active immunization with a conjugate of dopamine and bovine serum albumin on the development of an experimental MPTP-induced depressive syndrome and on the monoamine metabolism in the brain of rats]. AB - Active immunization with dopamine conjugated with bovine serum albumin (DA-BSA) or BSA with complete Freund's adjuvant (CFA) partly suppressed the development of the MPTP-induced depressive syndrome in rats preventing the appearance of "behavioral despair" symptoms: increase in immobility time and higher index of depression in forced-swim test. In DA-BSA-immunized rats the content of DOPA, DA, HVA, NA, and 5-HN in caudate putamen and that of NA in the frontal cortex was increased, while in BSA-immunized rats the content of 5-HT in both brain areas and that of DOPAC in the frontal cortex was decreased both in rats with reduced depressive syndrome and in saline control as compared with intact animals a day after the last drug injection. In DA-BSA-immunized rats with reduced depressive syndrome the increase in DA and 5-HT content in caudate putamen was less expressed and DOPAC content was lower than in saline control. In BSA-immunized depressive rats DA content in the frontal cortex was also reduced as compared to control. PMID- 10822849 TI - [The spike reactions of the motor cortex neurons of old rabbits to specific stimuli]. AB - Spike reactions of motor cortex neurons to tactile and electrocutaneous stimulation of a forelimb were studied in aged (6-7-year old) rabbits. As compared with young adult animals, the neuronal reactions to afferent stimuli were rarely recorded in the motor cortex of aged rabbits (66.7 and 50%, respectively). The activation manifested in increasing firing rate over its spontaneous level was less intensive than in young animals. The neuronal reactions of aged animals were characterized by the slower activation with longer latencies and slower development of spike responses. The parameters of slow activation could be partly corrected by the iontophoretic application of acetylcholine to the soma region. Neuronal inhibition recorded in the motor cortex of aged rabbits was not markedly changed compared to inhibition reactions in young animals. It is suggested that impairment of the functional state of dendrites in aging is responsible for the changes observed. PMID- 10822850 TI - [The late excitatory responses of the motor cortex neurons in the cat to stimulation of the pyramidal tract]. AB - Under conditions of partial suppression of GAMKA-dependent cortical inhibition in the motor cortex of anesthetized cats, a weak electrical stimulation of the pyramidal tract evoked the late slow (50-200 ms) excitatory reactions in the motor cortex neurons similar to those previously recorded under the same conditions in response to stimulation of the parietal cortex. This finding favors the proposal that the late excitatory component of the cortico-cortical response reflects the repetitive activation of cortical neurons due to excitation spread via the system of cortical recurrent excitatory collaterals. PMID- 10822852 TI - [In Process Citation] PMID- 10822851 TI - [Conceptual approaches to revealing the function of the structural organization of a neuronal network]. AB - Brain is an unexcelled instrument of perception and analysis of intensive information flows and decision making. Joint achievements of neurophysiology and morphology highlight the pathways between neuronal formations of different brain regions and their functions and fine organization of relationships within these formations. However, the principles of information transmission and signal processing in such structures remain hypothetical. The structure of the network executing the function of extrapolation or prediction of input signal values is discussed. Organization of neurons in this network is compared with relationships of neurons in the cerebellar cortex. It is suggested that the considered organization of neuronal relationships and the functions executed by the network are inherent for all brain formations including the brain cortex. PMID- 10822853 TI - Antimicrobial therapy in a historical perspective. PMID- 10822854 TI - The impact of antimicrobial resistance. PMID- 10822855 TI - Evolution and mechanisms for spread of antimicrobial resistance. PMID- 10822856 TI - Antibiotics in animal feed. PMID- 10822857 TI - Spread of antibiotic resistance from food animals to man. PMID- 10822858 TI - Spread of resistance from food animals to man: the French experience. PMID- 10822859 TI - Spread of resistance from food animals to man--the UK experience. AB - Since 1990 there have been dramatic increase in the occurrence multiply drug resistant strains of zoonotic pathogens causing infections in humans in many developed countries. Of particular note has been the epidemic spread of MR strains of S. typhimurium DT 104, which now appear to have an almost world-wide distribution. Within DT104 the increasing spectrum of resistance is of considerable concern, with strains with decreased susceptibility to ciprofloxacin increasing in incidence in the United Kingdom and also causing serious disease in humans in other countries. For campylobacters the incidence of ciprofloxacin resistant organisms is also increasing, with reports of such isolates from numerous countries throughout the world. For VTEC O157, although resistance is increasing, multiple resistance and resistance to ciprofloxacin remains rare. Drug resistance in food-borne pathogens is an unfortunate but almost inevitable consequence of the use of antimicrobials in food animals. Although for some pathogens--e.g., Campylobacter spp., the use of antimicrobials in human medicine is also an important factor (Smith el al 1999), it is the use of antimicrobials in food animals which has been a major factor in the development of decreased susceptibility to antibiotics such as ciprofloxacin in zoonotically-transmitted salmonellas. Such use is quite legitimate. However it is regrettable that recommendations such as propounded in 1992 in the UK by the Expert Group on Animal Feedingstuffs--the Lamming Committee, that any new antibiotics with cross resistance to those used in human medicine should not be used for prophylaxis in animal husbandry, were not accepted (Anonymous, 1992). Although the clock cannot be turned back, to combat the development of resistance to such important drugs as the fluoroquinolones it is hoped that a Code of Practice for their use in food animals will soon be internationally implemented. PMID- 10822860 TI - Role of milk and meat products as vehicles for antibiotic-resistant bacteria. PMID- 10822861 TI - Figures on quantities of antibacterials used for different purposes in the EU countries and interpretation. PMID- 10822862 TI - Effect of intervention on the occurrence of antimicrobial resistance. PMID- 10822863 TI - The effects of intervention on antimicrobial resistance. PMID- 10822864 TI - Sense and nonsense in surveillance programs. PMID- 10822865 TI - Farming of Atlantic salmon--an experience from Norway. PMID- 10822866 TI - Disease preventive methods as alternatives and complements to the use of antibiotics. PMID- 10822867 TI - Resistance to antibiotics--a threat to human health. PMID- 10822868 TI - Serious and fatal firearm injuries among children and adolescents in Alaska: 1991 1997. AB - STUDY OBJECTIVE: To describe demographics, causal factors, intent, and incident locations of serious and fatal firearm injuries among children and adolescents in Alaska, for the years 1991 through 1997. METHODS: Data from the Alaska Trauma Registry plus Vital Statistics death certificates were reviewed for a seven-year period (1991-1997). Data elements included are: intent (ICD 9-CM E-Codes and narratives); age group; region of incident; place of occurrence; alcohol or drug involvement; type of firearm used; and perpetrator. RESULTS: During the seven year study period, 222 children and adolescents ages 0-19 years were admitted to a hospital for a non-fatal firearm injury, plus 165 others received fatal firearm injuries. Of these 387 serious and fatal injuries, 34.9% (135) were determined to be unintentional, 36.4% (141) were suicides or suicide attempts, 23.3% (90) were homicide/assaults, 0.5% (2) were legal intervention, and for 4.9% (19) intent was unknown. Rates of serious and fatal firearm injuries per 100,000 youth for the six-year study period ranged from 14 in the Fairbanks North Star Borough and the Kenai Peninsula Borough to 105 in the Yukon-Kuskokwim Region. The statewide average for this period was 27.1 per 100,000 children and adolescents. CONCLUSIONS: Firearm injuries are a leading cause of serious and fatal injuries to children and youth in Alaska. This study suggests that many children and adolescents in Alaska who were injured by firearms, or who caused injury to other children or youth by firearms, had easy access to them. Efforts should be made to convince adults not to let children or at risk teenagers have unsupervised access to firearms, and to promote safe storage of firearms. PMID- 10822869 TI - Providing health coverage for Alaskans in need. Alaska's Division of Medical Assistance improves service to its customers. PMID- 10822871 TI - Struggle to resolve the oil company control issue has ended PMID- 10822870 TI - Diabetes care: a comparison of management systems. AB - During 1997, the Alaska Area Diabetes Program audited a sample of 837 charts from among the 1373 individuals receiving regular diabetes care at facilities serving predominantly American Indians/Alaska Natives. Charts were evaluated for the delivery of American Diabetes Association Standards of Care. Results of the audit showed a great deal of variability among facilities. Facilities which scored highly in the audit were more likely to use an organized multidisciplinary team approach that included coordinated clinic appointments with multiple providers on the same day, maintenance of a diabetes registry, proactive preclinic preparation, flow-sheet use, intensive individual nutritional counseling, a case manager or field clinic coordinator system with standing orders, and strong self management support. PMID- 10822872 TI - [Holistic view of internal medicine]. PMID- 10822873 TI - [Where is medical education going?]. PMID- 10822874 TI - [Retroviruses and opportunistic infections. VII International Congress. January 30-February 2, 2000, San Francisco, USA]. PMID- 10822875 TI - [Plasmapheresis combined with pharmacology in the treatment of lupus nephritis: a new therapeutic protocol]. AB - Systemic Lupus Erythematosus (SLE) is a pathology of unknown cause, characterized by tissue and cellular damage, secondary to production of autoantibodies and to deposition of immunocomplexes. Lupus nephritis is one of the most common complications of SLE. Our purpose is to propose a therapeutic protocol for patients suffering from Lupus nephritis. This consists of Plasmapheresis associated with pharmacological therapy so that dosage of immunosuppressors can be reduced and consequently also the side effects of the pharmacological therapy. PMID- 10822876 TI - Reactive oxygen species and antioxidant status in essential arterial hypertension during therapy with dihydropyridine calcium channel antagonists. AB - PURPOSE: To evaluate reactive oxygen species and antioxidant status in essential arterial hypertension during therapy with dihydropiridine calcium channel antagonists. PATIENTS AND METHODS: Fifteen patients, affected by essential arterial hypertension, were examined. They received once a day oral dihydropyridine calcium antagonists for 10 weeks: five patients received felodipine (5 mg), five amlodipine (10 mg) and five lercanidipine (10 mg). The levels of end products of lipid peroxidation, free radicals and hydroperoxides and total antioxidant capacity were determined in the plasma of all subjects before and during treatment. Values are expressed as mean +/- S.E. Systolic blood pressure decreased from 171 +/- 4 to 135 +/- 6 mmHg (p < 0.01) and diastolic blood pressure decreased from 99 +/- 5 to 82 +/- 3 mmHg (p < 0.01). Hydroperoxides and free radicals decreased from 321.3 +/- 8.96 to 247.9 +/- 8.69 units (p < 0.01) and the end products of lipid peroxidation decreased from 11.0 +/- 1.93 to 6.74 +/- 1.41 nmol/ml (p < 0.01). Total antioxidant capacity increased from 0.74 +/- 0.03 to 1.05 +/- 0.05 mmol/l (p < 0.01). RESULTS: Imbalance in the pro-oxidant-antioxidant equilibrium shifts in favour of antioxydant namely oxidative stress decreases. The calcium channel antagonists decrease peripheral arterial resistances and therefore decrease or abolish relative ischaemia, moreover decrease arterial pressure and therefore normalize parietal stress on endothelial cells. As a conseguence they act on two hypothesized mechanisms of oxidative stress in hypertension. CONCLUSIONS: Dihydropyridine calcium antagonists used in this trial seem useful in hypertension because they decrease oxidative stress, and normalize of pressure values. PMID- 10822877 TI - [Drug adverse effects in the elderly hospitalized with acute pathologies]. AB - OBJECTIVES: Data of Literature suggest that age is not an independent risk factor for adverse drug reactions (ADR), while there is evidence of a positive correlation between ADR and the number of drug taken. To investigate if that is true, we have examined the occurrence of ADR in elderly patients admitted to our Geriatric Department in the first nine months of 1999 for acute illness. MATERIALS AND METHODS: We have evaluated the occurrence of ADR in 510 patients more than 65 years of age (80 +/- 7.35 years). The adverse drug effect was worth considering when symptomatical or caused a change in the management of the patient (suspension or change of therapy, prolongation of staying in hospital). We used the algorithm of Naranjo (19) to estimate the probability that a drug caused an ADR. RESULTS AND CONCLUSIONS: In our study the frequency of ADR was 12.5%. Furosemide was responsible for 50% and digoxin for 8% of ADR. Among the patients taking furosemide, 12.5% had an ADR (generally low potassium). The frequency of ADR was correlated neither with the age of the subjects nor with the number of drug taken. It can be possible that the main cause of ADR is the type of drugs assumed by elderly. PMID- 10822878 TI - [Changes in guidelines for the therapy of non-nosocomial pneumonitis]. AB - In the USA therapeutic suggestions for a more appropriate treatment of community acquired pneumonias, are presently under revision. In this review we underline the difficulties encountered today, at the beginning of the year 2000, in the formulation of appropriate guidelines for this disease, taxed just about constantly by a mortality rate of nearly 8%. We emphasized the problem of bacterial resistance to multiple antibiotics and the frequency of polymicrobial pulmonary infections with an obvious worsening of the prognostic outlook. Directions about choice and use of some antibiotics have been provided, citing especially the fluoroquinolones and the more recent one, the moxifloxacine. A recommendation is given as well to pharmaceutical companies for them to provide better directives about the ideal modalities of administration of new antibiotics with the double aim of helping patients and also attempt to prevent onset of bacterial resistance. PMID- 10822879 TI - [Medullary carcinoma of the thyroid: diagnosis and therapy]. AB - Medullary thyroid carcinoma (MTC) originates in the thyroid C cells, or parafollicular cells, secreting calcitonin. It may be either sporadic or familial. Familial form can be isolated or expression of a multiple endocrine neoplasia type II. Mutations of the RET proto-oncogene have been identified in the germline DNA of patients with familial MTC syndromes. Genetic testing can identify patients affected by multiple endocrine neoplasia and familial MTC, allowing early diagnosis and possible cure. The initial treatment is surgical and the adequate surgery consists of total thyroidectomy. The treatment of occult or minimal disease can be curative. Plasma calcitonin measurements are excellent markers for post-operative follow-up. Imaging study can help to discover recurrent or metastatic disease. Adjunctive therapy includes radiotherapy and chemotherapy. Radiotherapy is reserved for bone metastases or for non resectable neck recurrences. Chemotherapy is reserved for patients with progressive MTC. Many chemotherapeutic regimens have been tried, results are controversial. PMID- 10822880 TI - [Diagnostic and therapeutic strategies in post-infarct]. AB - It is of primary importance for the clinical cardiologist to keep in mind the parameters allowing an adequate prognostic stratification in post-infarct patients in view of making the best diagnostic and therapeutic choices. A diagnostic strategy, based on a pathophysiologic approach, should evaluate four aspects: spontaneous and stress-induced ischemia, myocardial viability, and ventricular arrhythmias. Spontaneous ischemia has an undefined prognostic value, especially in the thrombolytic era; therefore it seems reasonable to perform invasive procedures in patients who are not stabilized by an adequate medical therapy or with large jeopardized areas. In asymptomatic patients, a provocative stress test allows a more articulated decisional iter. It is preferably to perform the test by the most physiological exercise EKG, together with the echocardiographic imaging, after the acute phase. It has a high negative predictive value, but a low positive predictive value. The detection of myocardial viability is frequently performed, mainly in patients with large post ischemic myocardial dysfunction. Among all the proposed methods, the echo dobutamine test mainly allows to estimate patients in whom revascularization may result in more benefit. The role of ventricular arrhythmias as an independent prognostic factor is debated and has to be always considered in relationship to other parameters, particularly left ventricular function. Regarding the therapeutic strategy, the indications from recent trials, related to antithrombotic drugs, beta-blockers, ACE-inhibitors, nitrates, Ca-blockers and antiarrhythmic drugs, are emphasized. PMID- 10822881 TI - [Clonidine hallucinations: description of a clinical case]. AB - Authors report a case of hallucinations related to clonidine treatment in an elderly man with hypertension and renal failure. The symptoms appeared shortly after treatment is starting. The hallucinations disappeared after clonidine withdrawal. Implications for clinical practice are evocated. PMID- 10822882 TI - [Evolution of chemotherapy of cancer. Conceptuals models]. PMID- 10822883 TI - Location of the glenoid fossa after a period of unilateral masticatory function in young rabbits. AB - Changes in glenoid fossa position and skull morphology after a period of unilateral masticatory function were studied. The right-side maxillary and mandibular molars in twenty-seven 10-day-old rabbits were ground down under general anaesthesia. The procedure was repeated twice a week, until the rabbits were 50 days old. Fourteen rabbits were then killed and 13 left to grow to age 100 days. Nine 50-day-old and sixteen 100-day-old rabbits with unmodified occlusions served as controls. Three-dimensional measurements were made using a machine-vision technique and a video-imaging camera. The glenoid fossa position become more anterior in both groups of animals subjected to molar grinding as compared with controls (P < 0.01 in the 50-day-old group and P < 0.05 in 100-day old group). In the 100-day-old group the right-side fossa was also in a more inferior position (P < 0.01). The glenoid fossa was more anteriorly located on the right than on the left side of individual animals in the group in which the right-side molars had been ground down (P < 0.001). PMID- 10822884 TI - A mathematic-geometric model to calculate variation in mandibular arch form. AB - A mathematic-geometric model was used to evaluate the variation of mandibular dental arch length with respect to the incisor inclination, but without modifying the intercanine width. In analytical terms, the equations of the curves representing the lower dental arch, before and after incisor inclination of 1 mm and of 1 degree, with controlled and uncontrolled tipping, were studied. The length of the mandibular dental arch changed in the parabolic arch form by 1.51 mm for each millimetre of incisor inclination with respect to the occlusal functional plane, by 0.54 mm for each degree of controlled tipping and by 0.43 mm for each degree of uncontrolled tipping. In the elliptical arch form (e = 0.78), it changed by 1.21, 0.43, and 0.34 mm, respectively, in the hyperbolic form by 1.61, 0.57, and 0.46 mm, in the circular form by 1.21, 0.43, and 0.34 mm, and in the catenary form by 2.07, 0.74, and 0.59 mm. The results show that by changing the arch form without modifying the dimension of the dental arch, different arch lengths can be gained for each millimetre of proclination. In addition, by controlled tipping an inter-incisive arch one-fifth longer than by uncontrolled tipping can be obtained. It would be advisable in orthodontic treatment planning to evaluate the type of dental arch, since the space available or the space required changes depending on the arch form and on the orthodontic tooth movement. PMID- 10822885 TI - Mixed dentition space analysis in a Thai population. AB - This study produced simple linear regression equations to be used for mixed dentition space analysis for males and females, and sexes pooled in a population living in northeastern Thailand. Measurements of teeth were made to within 0.01 mm on the dental casts of 215 boys and 215 girls (mean age 15.7 years). All dentitions were required to be free of any signs of dental pathology or anomalies. It was found that males had significantly larger teeth than females as represented by summations of mandibular incisor, canine, and premolar widths. ANOVA of regression indicated a close relationship between mandibular incisor summation and corresponding summations of canine and premolars. The low coefficients of determination (r2) of the regressions ranged between 0.29 and 0.42, and were higher for females than males, which might be attributable to the ethnic diversity of the sampled population. The regression equations produced predictions of mesio-distal width summations for maxillary and mandibular canine, and premolar arch segments that were slightly different from other reported Asian studies. Moyers' prediction tables at the 50th percentile were found to under estimate tooth size summation compared with the present investigation. The predictions from simplified regression equations matched well with those of this study for sexes pooled, and for males and females separately. PMID- 10822886 TI - Analysis of dental casts of 6-8- and 12-year-old Kenyan children. AB - Dental plaster casts of 97 6-8-year-old and 173 12-year-old Maasai, Kikuyu, and Kalenjin children were studied. The Kikuyu are Bantu, while Maasai and Kalenjin are Nilo-Hamitic subjects. The variables measured were palatal depth (PD) and length (PL); maxillary and mandibular anterior arc circumferences (AC1) and (AC2), respectively; posterior arc circumferences (PC1) and (PC2), inter-canine (CC1), and (CC2); inter-molar (MM1) and (MM2) distances, and mandibular length (ML). The data were analysed using SPSS package. The mean values of all the variables were generally higher in the males compared with the females and significant sex differences in the means (P < 0.01) were found in AC1, PC1, PC2, CC1, CC2, MM1, and MM2 in the 12-year-old subjects. The means of all variables, except PL, ML, PC2, and CC2, increased from 6 to 12 years of age and significant differences in the means for age were found in ML, AC1, PC2, PD, MM1, MM2, and CC1. Mean maxillary inter-molar distance increased with age by 0.17-0.34 mm in the three groups. Mean values of mandibular variables were highest in the Kikuyu, while maxillary variables were highest in the Maasai. The Maasai casts showed a marked decrease in CC2, PC2, AC2, and ML compared with the Kikuyu and Kalenjin. Ethnic and sex differences in the dental arches may be masked by anterior tooth positions that are influenced by the dento-alveolar complex and soft tissues. Corresponding mandibular and maxillary variables were strongly correlated and anterior and posterior arc circumferences were correlated with inter-canine and intermolar distances. Details of the norms for dental arch dimensions and changes with age may allow for appropriate assessment of dental occlusion and treatment planning for Kenyan children. PMID- 10822887 TI - Behaviour management needs for the orthodontic treatment of children with disabilities. AB - A displeasing dental appearance may have a significant emotional impact on an individual's well being. Although malocclusions occur more often in physically and/or mentally handicapped children than in normal children, the most severely handicapped patients are those least likely to receive orthodontic treatment. This investigation studied the modes of behaviour management used in the orthodontic treatment of disabled children, and the preferred criteria. The files of 49 disabled children were retrospectively evaluated. Two classification systems, the Frankl Behaviour Rating Scale (FBRS), and that of Owen and Graber were found to be unsuitable for determining the appropriate treatment modality. Five specific factors, frequently seen in disabled children, gag reflex, drooling, uncontrollable movements, inability to remain still, and the need for additional procedures, were graded and a scoring system was devised to include these factors within the assessment. This scoring system may be used to evaluate new patients and to assist in the choice of the appropriate behavioural management mode. PMID- 10822888 TI - Orthodontic treatment for disabled children: motivation, expectation, and satisfaction. AB - This study was designed to measure motivation for and expectations of proposed orthodontic treatment for disabled children, and to examine the level of satisfaction with the results of this treatment, in the eyes of the parents. A two-part questionnaire was sent to the parents of consecutively treated disabled children. The first part was sent to the parents of all the patients treated, while the second was only sent to those whose child had completed treatment. The response rate was over 90 per cent. The parents expected improvement in the child's appearance with a concomitant improvement in his/her social acceptance. These expectations from the treatment were found to be exaggerated, with only a minority of the parents claiming a marked improvement in their child's everyday functioning (four out of 27), or a significant social improvement (six out of 27). Nevertheless, most of the parents (26 out of 27) were satisfied with the treatment, and reported that 17 of the children themselves, who were aware of a change, considered it an improvement. A majority of the children understood the reasons for treatment, in the most general of terms. Close friends regarded treatment results as positive (20 out of 27). With only one exception, the parents stated that they would repeat the procedure, given the same set of circumstances, and all of them would recommend it for other disabled children. It may be concluded that even though orthodontic treatment in this groups of patients does not yield the desired social influence, the individual benefits from the treatment are worthwhile. PMID- 10822889 TI - Do oral appliances enlarge the airway in patients with obstructive sleep apnoea? A prospective computerized tomographic study. AB - This study evaluated the effect of an anterior mandibular positioning appliance (AMPA) on minimum pharyngeal cross-sectional area (MPCSA) in 32 conscious supine obstructive sleep apnoea (OSA) subjects. The change in MPCSA was measured using low dose computerized tomography, with and without an AMPA in situ. The results showed that the mean presenting respiratory disturbance index (RDI) was 26.6 events/hour, with a body mass index of 28.6 kg/m2 and mean age of 51.5 years. There was a statistically significant increase in MPCSA of 28.34 mm2 on appliance insertion (SD = 59.06 mm2; range -145 to +190 mm2; P = 0.011). The mean mandibular displacement was 5.73 mm (SD = 2.51 mm) in protrusion and 8.27 mm (SD = 4.51 mm) inferiorly. A poor correlation was found between the size of the mandibular displacement and the change in MPCSA (protrusion r = 0.268; inferiorly r = 0.240, P > 0.05). In conclusion, the AMPA significantly increased MPCSA, suggesting that it may be an effective therapy for OSA. There was, however, a wide but unpredictable individual variation of response. As a small number of patients may worsen in their condition with temporary mandibular advancement (TMA), it is essential that all patients treated with TMA should be investigated by polysomnography both before and after treatment. PMID- 10822890 TI - Classification of occlusion reconsidered. AB - Katz's quantitative modification of Angle's occlusion classification has been found to have a high intra- and inter-examiner agreement among orthodontists. In the present study an attempt was made to introduce a 'combined' system comprising Katz's modification and overjet/overbite millimetric measurements in order to attain a more meaningful and complete classification of malocclusion than is presently available. A group of 32 raters (16 orthodontists and 16 senior-year students) examined 14 study models twice, with an interval of at least 1 month between examinations. In total, 448 x 2 determinations were performed. The percentage agreement of the Angle, the modified and the 'combined' systems, as well as the performance of the orthodontists versus the students were compared using the paired t-test. The percentage agreement obtained by both orthodontists and students was highest for Katz's modification and lowest for Angle's method. The overjet/overbite measurements affected the agreement in Katz's modified technique. The orthodontists surpassed the students with respect to Angle's method (P = 0.025), whereas no statistically significant difference existed between orthodontists and students regarding Katz's modification or the 'combined' system. It is concluded that in view of the relatively low agreement in the 'combined' method, it cannot be recommended for clinical application. The Katz's modified method, on the other hand, may be a helpful supplement to Angle's classification. PMID- 10822891 TI - Perceived aesthetic impact of malocclusion and oral self-perceptions in 14-15 year-old Asian and Caucasian children in greater Manchester. AB - The aims of this study were to evaluate (i) the effect of ethnicity, social deprivation, and normative orthodontic treatment need on orthodontic aesthetic self-perception, self-perceived need for orthodontic treatment, and oral aesthetic impact of malocclusion; (ii) the effect of ethnicity, social deprivation, and gender on perceived orthodontic treatment need and use of orthodontic services; (iii) the influence of perceived oral aesthetic impact of malocclusion on perceived need and wish for orthodontic treatment; and (iv) whether orthodontic treatment experience influences perceived oral aesthetic impact of malocclusion. A stratified, random sample of 434 14-15-year-old children from schools in Manchester, UK, was obtained. Information was collected on orthodontic aesthetic self-perception and orthodontic treatment experience using a questionnaire. The former data were combined to form an Oral Aesthetic Subjective Impact Scale (OASIS). Normative orthodontic treatment need was measured with the Index of Orthodontic Treatment Need (IOTN). Children with higher clinical need for orthodontic treatment perceived themselves as worse off than their peers with lower need. More socially deprived children or those with high IOTN aesthetic component (AC) scores had a higher (i.e. more negative) aesthetic impact (OASIS) score. Asians and females had higher IOTN dental health component (DHC) scores, but a better aesthetic appearance than Caucasians and males. More deprived children were less likely to have received orthodontic treatment. Despite this, OASIS scores were similar between treated and untreated children. Untreated children who wished for orthodontic treatment had higher IOTN AC and OASIS scores. PMID- 10822892 TI - The prenatal cranial base complex and hand in Turner syndrome. AB - From early childhood, Turner syndrome patients have a flattened cranial base, maxillary retrognathism, and short hands. There are, however, no studies that show when these genotype-determined abnormalities occur prenatally. The purpose of the present study was to measure craniofacial profile and hand radiographs of second trimester foetuses with Turner syndrome and compare the results with similar measurements from normal foetuses. The subjects consisted of 12 Turner syndrome foetuses, gestational age (GA) varying between 15 and 24 weeks, and crown-rump length (CRL) between 108 and 220 mm. The mid-sagittal block of each cranium was analysed as part of the requested brain analysis (pituitary gland analysis). This block and the right hand from seven foetuses were radiographed, and the skeletal maturity of the cranial base complex, i.e. the cranial base and the maxilla, was evaluated from the profile radiographs. Shape and size measurements in the cranial base were performed, and compared with normal values according to cranial maturity and to CRL. The cranial base angle in Turner syndrome was greater and the maxillary prognathism was reduced compared with the normal group. The dimensions in the cranial complex and in the hand showed that the bone lengths and distances in relation to CRL were generally smaller compared with normal foetuses. This investigation showed that the abnormal shape of the cranial base complex and the short hands in Turner syndrome are present prenatally. PMID- 10822893 TI - Are orthognathic patients different? AB - This questionnaire-based study investigated the psychological profile of orthognathic patients prior to starting treatment and compared the findings with a control group of non-patients. Comparison of the data used multivariate multiple regression analysis where outcome variables and independent variables were studied simultaneously. Some differences were found in the psychological profile of the orthognathic patient. They displayed higher levels of state anxiety (P < 0.001), higher numbers of individuals in their social support network (P < 0.05), and lower body image and facial body image (P < 0.001). Self esteem was also found to be lower, but only at borderline levels of significance (P = 0.052). PMID- 10822894 TI - The JIM interview. Edward M. Kennedy, United States Senator. AB - Senator Edward M. Kennedy has represented Massachusetts in the United States Senate for thirty-six years. He was first elected in 1962 to finish the term of his brother, President John F. Kennedy. Since then, he has been elected to six full terms, and he is now the third most senior member of the senate. The efforts to bring quality health care to every American is a battle that Kennedy has been waging ever since he arrived in the Senate. Recent achievements include the Health Insurance Portability and Accountability Act of 1996, which makes it easier for those who change their job or lose their job to keep their health insurance, and the Children's Health Insurance Act of 1997, which makes health insurance more widely available to children through age 18 in all 50 states. A strong supporter of clinical research, Senator Kennedy cosponsored the Clinical Research Enhancement Act and has been a vocal advocate of stem cell research. He is currently the senior Democrat on the Labor and Human Resources committee in the Senate. PMID- 10822895 TI - Action in Washington surrounding allegations of fetal tissue for sale. PMID- 10822896 TI - NIH and FDA seek to increase oversight of gene therapy research. PMID- 10822897 TI - Novel human autoantibodies to phosphoepitopes on mitotic chromosomal autoantigens (MCAs). AB - BACKGROUND: Human autoantibodies to proteins of the mitotic apparatus have demonstrated clinical utility and usefulness as molecular probes for identification and characterization of novel autoantigens, as exemplified by autoantibodies to centromere proteins. In contrast, there have been very few reports of autoantibodies with reactivity to antigens located along mitotic chromosome arms, but not in interphase nuclei. The purpose of this study was to identify and characterize autoantibodies with reactivity to mitotic chromosomal antigens (MCAs) located exclusively on mitotic chromosome arms, and to determine if patients with these autoantibodies have common clinical features. METHODS: Routine immunofluorescence screening of serum samples referred for antinuclear antibody investigation over a 10-year period was used to identify autoantibodies to MCAs. MCAs were identified by exclusive immunofluorescence staining of mitotic chromosome arms with no staining of interphase nuclei. MCA-reactive sera were further characterized for patterns of staining on mitotic chromosome arms and sensitivities to chemical and enzymatic treatments, and for one of these sera, its ability to abrogate progression through mitosis when microinjected into cells. RESULTS: Of 60,000 sera screened for antinuclear antibodies by immunofluorescence, we identified three IgG autoantibodies reacting exclusively to MCAs. The anti-MCA autoantibodies did not react with condensed chromatin in spermatozoa or in apoptotic HeLa cells. Reactivity of all three sera was abrogated by treatment with protease, but not RNase, indicating that the MCAs are protein in nature and do not contain RNA epitopes. The three anti-MCA antibodies seem to react to three different antigens because they gave different patterns of staining of chromosome arms, reacted with chromosomes in different stages of mitosis, and displayed different sensitivities to treatment with DNase 1, salt, and phosphatases. Phosphatase treatment suggests that MCA1 and MCA2 contain serine/threonine phosphoepitope(s) and MCA3 tyrosine phosphoepitope(s). Loss of MCA2 reactivity to DNase 1 treatment and its retention after salt extraction suggests that it is a chromosomal scaffold protein. Sensitivity of all three MCAs to acid suggests that they are histone-like or histone-associated proteins. CONCLUSIONS: We report the identification of three novel MCA-reactive sera. Patient diagnoses included discoid lupus erythematosus, chronic lymphocytic leukemia, Sjogren's syndrome, and polymyalgia rheumatica. The reactivity of anti MCA antibodies with phosphoepitopes is likely to explain restriction of immunofluorescence staining to chromosome arms during mitosis. Microinjection of MCA1-reactive antibodies led to metaphase arrest, without any change in morphology of the mitotic spindle or metaphase chromosomes suggesting that MCA1 may have a role in sister chromatid separation. PMID- 10822898 TI - Fluvastatin reduces soluble P-selectin and ICAM-1 levels in hypercholesterolemic patients: role of nitric oxide. AB - BACKGROUND: Lipid-lowering therapy with 3-hydroxy-3-methylglutaryl-coenzymeA (HMG CoA) reductase inhibitors reduces the incidence of atherosclerosis-related cardiovascular events. Adhesion molecules, regulating interactions between vascular and circulating cells, may play a central role in the pathogenesis of atherosclerosis and related complications. In the present report we examined the impact of the HMG-CoA reductase inhibitor fluvastatin on plasma levels of P selectin and ICAM-1. METHODS: Plasma levels of P-selectin and ICAM-1 were determined using an enzyme immunoassay in 26 patients with type IIa hypercholesterolemia randomized to treatment with either fluvastatin (80 mg/d) or placebo in a double blind fashion for 12 weeks. RESULTS: Fluvastatin administration reduced either P-selectin (118 +/- 63 vs 81 +/- 36 ng/mL [-31%], P = 0.0015) or ICAM-1 (264 +/- 75 vs 228 +/- 68 ng/mL [-13.7%], P = 0.0033) levels. Fluvastatin also lowered urinary 11-dehydro-TXB2 (1396 +/- 536 vs 1009 +/- 378 pg/mg creatinine [-27%], P = 0.0015) and von Willebrand Factor levels (1456 +/- 716 vs 1203 +/- 527 U/L [-17.4%], P = 0.0275), and a direct correlation was observed between P-selectin and 11-dehydro-TXB2 levels (r = 0.588, P = 0.0033). Patients treated with fluvastatin displayed an increase in nitric oxide (NO) generation, evaluated with measurements of serum NO2-/NO3-, (4.7 +/- 1 vs 8.9 +/- 3.1) mumol/L [98%], P = 0.0046). Moreover, an inverse correlation was observed between NO2-/NO3- and P-selectin (r = -0.420; P = 0.0343), 11-dehydro-TXB2 (r = 0.511; P = 0.0106), or LDL (r = -0.742; P = 0.0002) levels. CONCLUSIONS: These results may provide novel biochemical basis for the beneficial clinical effects of HMG-CoA reductase inhibitors in hypercholesterolemia. PMID- 10822899 TI - Protein complexes involving alpha v beta 3 integrins, nonmuscle myosin heavy chain-A, and focal adhesion kinase from in thrombospondin-treated smooth muscle cells. AB - alpha v beta 3 integrins have been implicated in regulating vascular healing in animal models of arterial injury. Because the specific cellular events mediated by alpha v beta 3 integrins are not completely understood, we examined alpha v beta 3 integrin-dependent cytoplasmic events in cultured human smooth muscle cells (SMC) following treatment with thrombospondin-1 (TSP), a glycoprotein concentrated at sites of blood vessel injury. TSP treatment elicited a time dependent association of nonmuscle myosin heavy chain-A (NMHC-A) with alpha v beta 3 integrins. NMHC-A also associated with focal adhesion kinase (FAK) in TSP treated SMC. FAK, a nonreceptor kinase implicated in integrin-mediated signaling, was phosphorylated on tyrosine in growth-arrested SMC, but levels of tyrosine phosphorylation increased following treatment with TSP. To test whether NMHC-A was regulated by vascular injury, we examined expression in baboon brachial arteries. In uninjured arteries, NMHC-A staining was present in the media. In arteries injured by balloon withdrawal, medial NMHC-A expression was increased with intense staining at specific sites. In summary, heteromeric protein complexes involving alpha v beta 3 integrins, NMHC-A, and FAK form following treatment of human SMC with TSP. These results suggest that the formation of protein signaling complexes is one mechanism whereby alpha v beta 3 integrins influence intracellular signaling pathways. PMID- 10822900 TI - Infection-susceptibility alleles of mannose-binding lectin are associated with increased carotid plaque area. AB - The MBL gene, encoding mannose-binding lectin, determines interindividual variation in susceptibility to certain infectious agents, such as Chlamydia pneumoniae. We examined whether infection-susceptibility alleles of MBL, called "non-A alleles," would be associated with increased carotid plaque area (CPA), an intermediate phenotype of atherosclerosis. In 164 subjects, we measured CPA with 2-dimensional ultrasound. We also determined traditional atherosclerosis risk factors and genotyped all subjects for MBL codons 52, 54, and 57. We used ANOVA to determine sources of variation for CPA and tested the hypothesis that the presence of a single MBL non-A "infection-susceptibility" allele was associated with increased CPA; 45.7% of subjects had at least one non-A allele. ANOVA showed that CPA was significantly associated with MBL genotype, age, smoking, hypertension, and hyperlipidemia (P < 0.05). When MBL was used as the sole independent variable in the regression analysis, the association with CPA was even more significant (P = 0.009). Subjects with at least one MBL non-A allele had significantly higher CPA than subjects homozygous for the MBL A allele and were significantly more likely to have CPA in excess of the sample median. Thus, infection-susceptibility alleles of MBL were associated with increased CPA in this study sample; these alleles may be a determinant of interindividual differences in atherosclerosis risk. PMID- 10822901 TI - Effect of aging on the arginine-vasopressin response to physostigmine and angiotensin II in normal men. AB - The effect of age on the stimulatory control exerted by cholinergic- and angiotensin II (ANG II)-mediated neurotransmission on arginine vasopressin (AVP) secretion was evaluated by measuring and comparing the AVP responses to the administration of either the cholinesterase inhibitor physostigmine (13.5 micrograms/kg in 50 mL normal saline infused in 10 minutes) or ANG II (increasing doses of 4, 8, and 16 micrograms/kg/min, each dose for 20 min) in 8 younger (23 37 years), 8 middle age (42-60 years), and 8 older (63-79 years) healthy male subjects. Both drugs induced significant increments in plasma AVP levels in the youngest group, with mean peak levels 4.8 times higher than baseline at 20 minutes after the beginning of physostigmine infusion and 1.5 times higher than baseline at 60 minutes after the beginning of ANG II infusion. Similar responses were observed in the middle age group. Basal AVP levels in older subjects were similar to those observed in the other groups. However, the AVP increases induced by physostigmine (mean peak was 9 times higher than baseline) and ANG II (mean peak was 2.2 times higher than baseline) were significantly higher in the oldest group than in the other groups. These data suggest age-related enhancement of the stimulatory regulation exerted by cholinergic- and ANG II-mediated neurotransmission on AVP secretion. PMID- 10822902 TI - Time-dependence of survival predictions based on markers of HIV disease. Swiss HIV Cohort Study. AB - OBJECTIVE: To determine whether the ability of baseline clinical stage, viremia, and CD4 cell counts to predict mortality in HIV-1-infected patients changes with duration of follow-up. METHODS: Three hundred ninety-four patients were followed for an average of 29 months by the Swiss HIV Cohort Study, a practice-based registry of HIV-1-infected patients in Switzerland. Predictor variables were the baseline clinical stage, CD4 cell count, circulating HIV-1 RNA level, and the RNA/CD4 ratio; the outcome was death. The changes in relative risks of death over time were examined using survival models that extend the Cox model to allow for nonproportionality of hazards. RESULTS: During 949 person-years of follow-up, 169 patients died (mortality rate 17.8 per 100 person-years). Compared with clinical stage A, patients in stages B and C at baseline had much higher mortality rates in the subsequent year. The prognostic ability of stage C decayed over time (P = 0.03). By contrast, the relative risks associated with a 2-fold difference in CD4 counts remained remarkably stable, at approximately 0.6 (P = 0.81 for the time dependence test). Relative hazards associated with a 10-fold difference in HIV RNA per milliliter and in HIV RNA per CD4 cell both tended to increase over time, but this trend failed to reach statistical significance (P = 0.21 and P = 0.08, respectively). CONCLUSIONS: Time-dependence patterns of prognostic ability varied widely among predictors, displaying gradual decay (clinical stage), stability (CD4 cells), and a trend to progressive increase (viremia). These results may affect clinical monitoring of HIV-infected patients and the interpretation of cohort studies of HIV-infected patients. PMID- 10822903 TI - The effects of physical training of functional capacity in adults. Ages 46 to 90: a meta-analysis. AB - BACKGROUND: There is a progressive decline in the functional capacity (FC) of the cardiovascular system with aging. This deterioration is reflected in a decrease in the maximal oxygen consumption (VO2max). The purpose of this quantitative review was to determine the effects of various components of the exercise prescription on FC in older individuals (ages 46-90 years). METHODS: This study synthesized the results of 27 studies by meta-analysis, which generated a total of 34 effect sizes. Each effect size represented an independent measure of the impact of physical training on maximal oxygen consumption (VO2max). A total of 720 subjects were included in this review. The studies were coded according to intensity, session duration, length of training and mode of exercise. RESULTS: A significant difference was found (p < 0.001) in FC between studies in which the intensity of exercise was > or = to 80% of VO2max compared to those with training intensities of 60-75% of VO2max. A significant difference was found (p < 0.002) for exercise duration; namely, a duration of > or = 30 minutes produced significantly greater improvements in VO2max when compared to an exercise duration less than 30 minutes. There was no significant difference reported in FC between studies that lasted 15 or more weeks in duration compared to those that lasted less than 15 weeks. Finally, there was no significant difference reported in FC in studies that utilized a walk/jog training mode compared to those which utilized cycle ergometry. CONCLUSIONS: Despite the inevitable decline in VO2max with aging, exercise training imparts favorable adaptations in FC in individuals well into their seventh and eighth decades of life. PMID- 10822904 TI - Kinetics of oxygen uptake during decremental ramp exercise. AB - BACKGROUND: The purpose of the present study was to examine the kinetics of oxygen uptake (VO2) during decremental ramp exercises after incremental ramp exercise above the anaerobic threshold. METHODS: Triangle exercise consisting of incremental and decremental exercises, and trapezoid exercise in which a constant load was inserted between incremental and decremental exercises were carried out. RESULTS: Blood lactate (La) increased in incremental phases during triangle and trapezoid exercises, and showed a further increase at constant work during trapezoid exercise. It significantly decreased during decremental exercise. Initial VO2 level during decremental exercise was significantly higher in trapezoid exercise than in triangle exercise. The VO2 level then exponentially decreased and later showed a linear decrease during both exercises. These VO2 kinetics were expressed by an approximate equation. The time constant expressing the exponential manner was about one minute on average in both exercises. The slope of the linearly decreasing phase of VO2 was significantly related to La at 100 watts during the decremental exercise. CONCLUSIONS: VO2 kinetics during decremental exercise after incremental exercise above the anaerobic threshold consists of at least three components: initial level, exponentially decreasing phase, and linearly decreasing phase. PMID- 10822905 TI - Improvement of motor abilities in pubertal girls. AB - BACKGROUND: Purpose of the study was to test the hypothesis that in the course of sexual maturation possibilities for accelerated development of motor abilities are triggered in girls. METHODS: A cross-sectional study was conducted on 77 healthy 11- to 14-year-old girls, grouped according to Tanner's 5-stage scale of sexual maturation. Motor abilities were assessed with the aid of 20 m dash, 4 x 9 m shuttle run, standing long jump, squats in 30 sec, sit ups in 30 sec, trunk forward flexion, Cooper 12-min running test, and Harvard step-test. RESULTS: Performance in the shuttle run, standing long jump and trunk forward flexion improved in correlation with sexual maturation stage. When consecutive maturation groups were compared, in standing long jump and trunk forward flexion significant differences were found between sexual maturation stages II and III, in shuttle run between stages I and II. The significant main effect of sexual maturation was confirmed with the aid of MANOVA. 23% of variance in the results of trunk forward flexion, 17% in standing long jump, and 10% in shuttle run were attributable to maturation differences. For the same three motor tasks 4%, 8%, and 15%, were respectively attributed to age differences. Differences between maturation groups disappeared when the results of shuttle run and standing long jump were adjusted with the aid of ANCOVA for age or height as the covariate, but persisted after results were controlled for body mass. CONCLUSIONS: The results support the tested hypothesis. Critical for improvement of agility is reaching maturation stage II, and for increase of leg muscle explosive strength and trunk flexibility, reaching stage III. PMID- 10822906 TI - Reliability of jumping performance in active men and women under different stretch loading conditions. AB - BACKGROUND: To determine the reliability of squatting jumps (SJ), counter movement jumps (CMJ) and drop jumps (DJ) tests, as well as the reliability of the optimal dropping height during drop jumping. METHODS: Jumping performance was assessed in 8 male and 9 female physical education students. Their age, weight and height (mean +/- SD) were 23.9 +/- 2.1 years, 72.0 +/- 12.1 kg, 174.3 +/- 10.4 cm, and 23.1 +/- 2.0 years, 54.8 +/- 4.9 kg, 160.1 +/- 5.0 cm for the males and females, respectively. The jumping performance was determined on six different testing days. On each testing day, squatting jumps (SJ) and counter movement jumps (CMJ) were performed as well as drop jumps (DJ) from heights between 20 and 100 cm. The dropping height given the maximum attained height was registered as the optimal dropping height (ODH). After a 15 min rest period, a 30 sec hopping test (HT) was performed and the mean height attained (MHT) as well as the number of jumps executed (NHT) were recorded. The height attained was computed from the flight time, which was measured with a digital timer (+/- 0.001 sec) connected to a resistive platform. RESULTS: The pooled coefficients of variation in percentage were 5.4 (SJ), 6.3 (CMJ), 6.2 (DJ), 31.9 (ODH), 3.1 (NHT) and 6.7 (MHT). A parabolic relationship between dropping height and attained height was found (r = 0.39-0.43, p < 0.001). The ODH was 48.2 +/- 14.0 cm and 62.9 +/- 21.3 cm for females and males, respectively (p < 0.05). Multiple regression analysis showed than ODH can be predicted from the SJ with a standard error of 9 cm. CONCLUSIONS: The variability of the assessment of jumping performance is similar to that reported for other variables used in the assessment of physical fitness. In contrast, the assessment of the optimal dropping height is less reliable. PMID- 10822907 TI - The effects of an application of suncream on selected physiological variables during exercise in the heat. AB - BACKGROUND: The purpose of this study was to investigate the effects of an application of suncream on mean skin temperature (MST), rectal temperature (Tre), oxygen consumption and heart rate during exercise in the heat. METHODS: Twenty two male subjects underwent two steady state exercise tests (55% of maximum oxygen consumption, mean work rate 153 watts) for 40 minutes (following a 5 minute warm-up), under two treatment conditions at the same ambient temperature (32 degrees C, 54% relative humidity). One treatment involved wearing an application of suncream (SC) while the other treatment did not (NSC). Suncream was applied in the amount of 30 ml/m2 of body surface area. RESULTS: Results of this study indicated that an application of suncream significantly reduced MST, on average. 31 degrees C, during exercise in the heat (p = 0.03). However, further statistical analysis revealed that this was a function of a significant interaction effect (p = 0.001) (main effect is sunscreen; interaction effect sunscreen plus temperature). MST was lower for the SC condition versus NSC over all time measurements. Results also showed a significant difference in the mean core to skin temperature gradient (p = 0.001), with a greater gradient existing in the SC condition. There did not appear to be any effect of suncream on ventilation, rectal temperature, oxygen consumption, lactate levels, sweat loss, percent plasma volume change, rating of perceived exertion, or respiratory exchange ratio. Interestingly, although not statistically significant (p = 0.18), rectal temperature remained lower, on average 0.1 degree C, under the SC condition for every measurement throughout the exercise session. CONCLUSIONS: In conclusion, results of this preliminary study suggest that an application of suncream does not impair heat dissipation as indicated by a decreased MST and increased core to skin thermal gradient. PMID- 10822908 TI - Pre-exercise carbohydrate and fluid ingestion: influence of glycemic response on 10-km treadmill running performance in the heat. AB - BACKGROUND: The purpose of this study was to determine the influence of ingesting solutions containing mixtures of carbohydrate (CHO) types on pre-exercise glycemic response, exercise-induced hypoglycemia, metabolic responses, and 10-km treadmill running performance in a warm environment. METHODS: Ten trained runners completed 6, self-paced 10-km treadmill runs one hour after ingesting 900 ml of one of the following test solutions: a water placebo (WP), an 8 g 100 ml-1 high fructose corn syrup solution (HFG; 72 g CHO), a 6 g 100 ml-1 glucose solution (GLU; 54 g CHO), a 6 g.100 ml-1 sucrose/glucose mixture (SUG; 54 g CHO), or banana with water to equal 900 ml (BAN; approx. 54 g CHO). The sixth condition was 675 ml of an 8 g.100 ml-1 HFCS solution (LFG; 54 g CHO). Blood samples were taken prior to ingestion and every 15 min during rest and at 15 and 30 min, and at the end of the 10-km run. Blood was analyzed for glucose (BG) insulin (IN), glycerol, lactate, and percent change in plasma volume. Urine volume during the 1 hour of rest and change in body mass during exercise were also determined. RESULTS: A significant (p < 0.05) correlation (r = -0.684) was seen between the pre-exercise glycemic response (PEGR = area under the resting BG curve) and the change in BG from pre-EX to 15 min of exercise. BG at 15 min of exercise was significantly higher in the WP (5.22 mM) versus the other conditions (HFG = 3.32, LFG = 3.91, GLU = 3.38, BAN = 3.74 & SUG = 3.63 mM). Pre-exercise IN was lower in the WP (6.54 U ml-1) condition versus the other conditions (HFG = 22.1, LFG = 16.2, GLU = 23.3, BAN = 18.8 & SUG = 12.8 U.ml-1). Ten km performance times were not different (WP = 41.87, HFG = 41.66, LFG = 41.79, GLU = 41.65, BAN = 41.53, and SUG = 41.75 min). A significantly greater body mass loss occurred due to urine production during the 60 min of rest in the WP compared to the other conditions. The degree of exercise-induced decline in blood glucose was related to the PEGR; however, the decline in BG did not affect 10-km running performance. In addition, there were no differences in the metabolic responses during exercise between the different CHO types, nor did the type of CHO influence running performance. Finally, the presence of CHO and/or electrolytes in the hydration solutions produced a better fluid retention during the 60-min pre-exercise rest period compared to water. CONCLUSIONS: The results confirmed that if a competitive athlete consumed a breakfast prior to ingesting a CHO-electrolyte beverage, a practice that is common, the glycemic responses may be different. PMID- 10822909 TI - Effect of ubidecarenone oral treatment on aerobic power in middle-aged trained subjects. AB - BACKGROUND: Coenzyme Q10 (CoQ10) plays an important role in oxidative mithocondrial phosphorylation and prevents lipid peroxidation in biological membranes. During sustained physical exercise, reactive oxygen species (ROS) production increase through several mechanism; one of them is the purine nucleotide cycle activation by shifting xanthine-dehydrogenase to xanthine oxidase during AMP breakdown. The aim of this study was to evaluate the effect of CoQ10 treatment on aerobic power. METHODS: EXPERIMENTAL DESIGN: according to a single blind study design, 28 health male cyclists were randomized into two groups (CoQ10 or placebo) and remained on treatments for eight weeks; there were 5 drop-outs and only 23 subjects were completely evaluated. Before and at the end of the eight weeks, cyclists underwent cardiopulmonary exercise testing. MEASURES: a software system performed the necessary calculations to obtain the following parameters: oxygen uptake, CO2 production, minute ventilation, oxygen ventilatory equivalent, carbon dioxide ventilatory equivalent, oxygen pulse. Finally oxygen peak and anaerobic threshold were determined. Moreover blood inosine, hypoxanthine, xanthine, lactate and CoQ10 levels were measured before and immediately after each test. RESULTS: The results of this study showed that at the end of the eight weeks there was no difference between the two groups concerning physiological and metabolic parameters, but muscular exhaustion was reached at higher workloads in the CoQ10 group. CONCLUSIONS: In our experience ubidecarenone oral treatment does not improve aerobic power. The little improvement of tolerance to higher workloads may be due to the antioxidant activity of CoQ10. PMID- 10822910 TI - Chest, waist, and hip circumference estimations from stereo photographic digital topography. AB - BACKGROUND: Stereo photographic digital topography (SPDT) is a new development of potential value for the determination of anthropometric circumference measurements of the chest, waist, and hip. The purpose of this study was to evaluate the reliability and validity of SPDT. METHODS: Subjects included 148 healthy female volunteers (age 21 to 51). The same-day test-retest reliability of SPDT was established by comparing initial measurements with a repeat. Validity was established by correlating SPDT measurements with criterion measures from traditional tape-measure assessment. RESULTS: Reliability coefficients of 0.99 were obtained for chest, waist, and hip circumferences. Further, high indices of validity were recorded for girths of the chest (r = 0.97, SEE = 2.57 cm), waist (r = 0.94, SEE = 3.16 cm), and hip (r = 0.96, SEE = 2.29 cm). CONCLUSIONS: High reliability and validity, low cost, ease of use, and comfort for subjects make SPDT a viable alternative to traditional anthropometric evaluation techniques. PMID- 10822911 TI - The relationship of hardiness, sense of coherence, sports participation, and gender to perceived stress and psychological symptoms among college students. AB - BACKGROUND: This study addresses the issue of why under conditions of stress some people stay physically and psychologically healthy while others become ill. Being able to deal with stress, to cope with the pressures of daily life, and yet stay healthy, is seen as a function of such factors as physical health, psychological health, constitutional predisposition, social support, exercise habits, and personality. METHODS: This study examined the moderating effects of the personality constructs of hardiness and sense of coherence, sports participation (college varsity athletes and college nonathletes), and gender on the relationship between perceived stress and psychological symptoms. College varsity athletes (n = 135) and college nonathletes (n = 135), all undergraduates at New York University, completed four questionnaires: Hardiness Scale, Sense of Coherence Scale, Daily Hassles Scale, and Profile of Mood States. Participants also completed a background questionnaire providing basic demographic data. Psychological symptoms and perceived stress were the criterion variables: hardiness, sense of coherence, sports participation, and gender were the predictor variables. Correlational analyses were applied to the resulting data and used to answer and to test the research hypotheses. RESULTS: There was a significant positive correlation between perceived stress and psychological symptoms among college varsity athletes and college nonathletes. There was a significant positive correlation between the personality scales of Hardiness and Sense of Coherence for both college varsity athletes and college nonathletes. When controlling for gender, college varsity athletes scored significantly higher on hardiness, scored slightly higher on sense of coherence, and reported significantly less perceived stress and significantly fewer psychological symptoms than the college nonathletes. Comparing by gender, no statistically significant mean differences were found on the four main variables. A significant negative correlation was observed between personality hardiness and perceived stress and psychological symptoms for the total sample. However, the college nonathletes subsample showed no significant effect between hardiness and perceived stress. CONCLUSIONS: Finally, there was a significant negative correlation between sense of coherence and perceived stress and psychological symptoms for both the college varsity athletes and college nonathletes. PMID- 10822912 TI - Caffeine use in sports. A pharmacological review. AB - Caffeine is the most widely ingested psychoactive drug in the world. As many know, chronic use of caffeine leads to dependence, tolerance, drug craving, and upon abrupt cessation unpleasant withdrawal symptoms. Thus, caffeine fulfills pharmacological criteria by which agents are classified as drugs of abuse. Nevertheless, its use is legal and only at high, but readily attainable, levels is it banned from sport. Its use is widespread by athletes as young as 11 years of age who are seeking athletic advantage over fellow competitors. It is likely that its use will not decline any time soon because it is inexpensive, readily available, medically quite safe, socially acceptable, and by most measures legal. However, at levels allowed in sport, caffeine through its wide-ranging physiological and psychological effects increases endurance in well-trained athletes. If the goal of drug-testing and education programs in sport is to protect the health of athletes, prevent unfair advantage (cheating) and encourage ethical behavior then it seems obvious that the allowable levels of caffeine ingestion should be decreased. The alternative is to continue with policies designed largely to punish only those that get caught. PMID- 10822913 TI - Markers of hydration status. AB - This paper reviews the literature, describes and discusses methods by which whole body hydration status can be determined in humans. A method of determining whether or not an individual is hypohydrated is of particular significance in an exercise situation as even moderate levels of hypohydration have a negative impact on exercise performance. Inspection of the published literature indicates that a number of methods have been used to determine hydration status. Body mass changes, urinary indices (volume, colour, protein content, specific gravity and osmolality), blood borne indices (haemoglobin concentration, haematocrit, plasma osmolality and sodium concentration, plasma testosterone, adrenaline, noradrenaline, cortisol and atrial natiuretic peptide), bioelectrical impedance analysis, and pulse rate and systolic blood pressure response to postural change are discussed. The urinary measures of colour, specific gravity and osmolality are more sensitive at indicating moderate levels of hypohydration than are blood measurements of hematocrit and serum osmolality and sodium concentration. Currently no "gold standard" hydration status marker exists, particularly for the relatively moderate levels of hypohydration that frequently occur in an exercise situation. The choice of marker for any particular situation will be influenced by the sensitivity and accuracy with which hydration status needs to be established together with the technical and time requirements and expense involved. PMID- 10822914 TI - Revalidation: the catalyst for change in continuing professional development? AB - Remarkably, only five years ago, hospital specialists were under no formal obligation to record participation in Continuing Medical Education (CME). No one seriously doubted that such education took place, but the system had never been challenged. In recent years, and with apparently ever increasing pace, all this is changing. Traditional, didactic, lecture based teaching of undergraduates is slowly vanishing from many undergraduate curricula. Postgraduate hospital training has been brought into line with Europe so that, within 7 years of full registration, it is theoretically possible to become a consultant. The Royal Colleges were charged with developing and monitoring a structured system of CME and chose to use the "points" system of which there was experience elsewhere. Around 70% of surgeons returned their annual CME forms and almost all met the agreed minimum points total. As a meaningful educational exercise which impacted favourably on patient care the points system was impossible to defend vigorously, and was ignored by an important minority. More recently still, our entire profession has been exposed to regular and fundamental criticism. There was a real risk that we would lose the right to self regulation. The GMC has now announced that every doctor will be required to undergo a process of appraisal that will lead to revalidation, allowing the doctor to remain on the national register. A full consultative process is now in place, with the Royal Colleges being required to implement a new system of annual review which will allow profiling of every surgical practice. While it is possible react to these changes with a combination of resentment and paranoia, the profession should grasp this opportunity for change. The sensible introduction of personal portfolios should allow us to maintain and improve standards of care, and allow us to demonstrate these standards to our peers, our employers and our patients. We may yet look back on clinical governance and revalidation as the catalysts which brought about constructive change to a potentially complacent profession. PMID- 10822915 TI - The management of sigmoid volvulus. AB - The epidemiology and clinical pattern of sigmoid volvulus are well defined. Although clinical manifestations of acute volvulus are often clear-cut, diagnostic doubt is not uncommon and, if gangrene supervenes, mortality rises appreciably. While gangrene requires resectional surgery, the management of the viable colon related to a volvulus episode has a variety of options. These, particularly non-resectional alternatives, require more critical reappraisal in the light of advances in minimally invasive techniques. PMID- 10822916 TI - Flap repair: an effective strategy for minimising sexual morbidity associated with the surgical management of vulval intra epithelial neoplasia. AB - OBJECTIVE: To assess the sexual function after local excision and flap repair for symptomatic vulval intraepithelial neoplasia (VIN). STUDY DESIGN: A retrospective analysis of five sexually active women who had persistent, symptomatic VIN diagnosed in a dedicated tertiary referral vulval clinic and treated with local excision and flap repair. Sexual function was assessed using a modified version of the Sabbatsberg Sexual Self-Rating Scoring (SRS) system. RESULTS: The mean age of the cohort was 32 years (range 30 to 51). Four patients had previously been unsuccessfully treated with local excision. Follow up ranged from 5 to 33 months. The SRS scores were 90, 90, 81, 72 and 25. Endogenous depression may explain this last score. There has been no recurrence of VIN. CONCLUSION: In sexually active women with symptomatic VIN III, a flap reconstruction should be considered in addition to local excision as the treatment of choice. PMID- 10822917 TI - Mummy 1911-210-1. AB - An ancient Egyptian mummy from the collections of the National Museums of Scotland was examined using computerised tomography (CT) scanning as part of the NMS mummy project. A facial reconstruction was produced from the CT scans for comparison with a painted 'portrait' which covers the face of the wrappings. The scans indicated the mummified wrapped body of an adult male 1.65 m tall with excellent preservation of the body. An exact replica, translucent model of the skull was created from the CT scans and standard cephalometric analysis undertaken. The facial features were reconstructed onto a plaster model of the skull from known tissue depths using terracotta clay. The resultant face was compared with the portrait and an extremely close match obtained, suggesting this was an individual portrait painted around the time of death. PMID- 10822918 TI - Laparoscopic anterior fundoplication. AB - Laparoscopic fundoplication has become the standard surgical method of treating gastro-oesophageal reflux disease. Although Nissen total fundoplication is the most commonly performed procedure, partial fundoplication, either anterior or posterior, is becoming more acceptable because of a suggested lower risk of long term side effects. This article describes a technique of laparoscopic anterior fundoplication. PMID- 10822919 TI - Hepatic malignancy: challenges and opportunities for the surgeon. AB - The management of hepatic malignancy has changed from a previously nihilistic approach to a more positive one. Every patient should be considered for curative surgical resection, either at first presentation or after cytoreductive treatment. The surgeon is faced with the challenge of adapting modern surgical techniques to increase resectability rates. Although the majority of patients are unsuitable for resection, effective palliation can be achieved using a wide variety of techniques. The rational implementation of these palliative modalities requires a multidisciplinary approach which provides the surgeon with the opportunity to play a key role in the ongoing management of these patients. PMID- 10822920 TI - Nuclear medicine in oncology. AB - The role of the nuclear medicine department in the diagnosis and management of malignant disease is constantly evolving. For example, the development of new radiopharmaceuticals, such as indium-111-labelled octreotide (pentetreotide), has enabled more specific imaging for the diagnosis of gastroentero-pancreatic (GEP) endocrine tumours to be carried out. New techniques such as sentinel lymph node localisation and biopsy now allow less invasive tumour staging. Advances in chemotherapy necessitate careful monitoring of the effects of chemotherapeutic drugs on normal organ function during follow-up, and tumour specific radiopharmaceuticals are being used for non-invasive, targeted tumour therapy. Nuclear medicine investigations should always be interpreted in the knowledge of results of other imaging investigations if they are to have greatest impact on patient care. PMID- 10822921 TI - Improving the mandibular reconstruction plate: technical innovation. AB - The mandibular reconstruction plate can be improved by constructing a shaped acrylic spacer secured with self-tapping screws. It is removed to leave space for bone grafting. PMID- 10822923 TI - Screening for early prostate cancer: what is the problem? AB - Screening has been applied to normal populations to detect early cancers. Prostate cancer is very common and can be detected at an early stage. Screening has not been introduced in the UK for this disease and the possible reasons for this are discussed. There is a strong case for evaluation of screening programmes for this disease. PMID- 10822922 TI - Internet mailing lists: a primer. AB - The widespread availability of new electronic sources of information has exponentially increased the amount of information available to the surgeon in recent years. These new sources of information come in various forms, each of which have their own rules and regulations. This article focuses on Internet mailing lists. The practicalities of mailing lists are discussed, with guidelines to help users find lists of interest, join and contribute to, and use this medium effectively while minimising the time spent on it. Pointers to starting one's own list are provided for the motivated reader. PMID- 10822924 TI - Metastasising dermatofibroma or dermatofibroma-like dermatofibrosarcoma protuberans? AB - Dermatofibroma is a relatively common cutaneous fibrohistiocytic tumour with a marked tendency to recur locally. It is almost invariably benign. In a review of the world literature there was one report of two patients with dermatofibroma with histologically documented metastases. In both of these there was local recurrence and subsequent metastasis to lymph nodes and haematogenous spread to the lungs. We present a case of cellular dermatofibroma of the lower limb with local recurrence and metastasis to the abdominal wall, posterior chest wall and both lungs 30 months after excision of the primary lesion. The clinical and pathological features of the case are presented to emphasis the need for follow up of patients with dermatofibroma which present any unusual histological features. PMID- 10822925 TI - Mycobacterium tuberculosis presenting as sternal osteomyelitis. AB - A 54-year-old female presented with a presternal abscess and developed axillary lymphadenopathy. Imaging confirmed the presence of sternal osteomyelitis. The osteomyelitis was cured by resection and muscle flap reconstruction. Although tuberculosis was suspected, the organism was only cultured after the fourth surgical procedure. Surgeons should be aware that negative microbiology does not exclude a diagnosis of Mycobacterium tuberculosis. PMID- 10822926 TI - Monitoring the critically ill patient. 1999; 45(6): 386-93. PMID- 10822927 TI - Lateral preputioplasty for phimosis. 1999; 45(5): 210-12. PMID- 10822928 TI - A new method of ileostomy formation following surgery for colonic obstruction. 1999; 44(6): 378. PMID- 10822929 TI - Local anaesthesia: to warm or alter the pH? A survey of current practice. 1999; 44(3): 167-71. PMID- 10822930 TI - Road traffic accidents. PMID- 10822931 TI - Bioelectrical impedance analysis of the body composition of Nigerian children with sickle cell disease. AB - We used bioelectrical impedance (BIA) to investigate the body composition of children with sickle cell disease (SCD) in northern Nigeria. A total of 48 children with SCD and 51 controls between 3 and 20 years of age were studied. A significant difference was found in the weights of male subjects over the age of 10 years compared to controls (p = 0.01), but not in height. Significant differences were also observed for SCD males in the 10-18-year-old age range in body mass index (p = 0.001), fat free mass (p = 0.001), per cent fat free mass, (p = 0.02), body fat (p = 0.02), and per cent body fat (p = 0.02). No significant differences in any of these parameters between SCD subjects and controls were obtained for males under the age of 10 years. There were no significant differences in the height, weight, body mass index, or fat free mass for female SCD subjects compared to controls over the age range we studied. However, there were significant differences in the per cent fat free mass (p = 0.006), body fat (p = 0.025), and per cent body fat (p = 0.01) for female SCD subjects over the age of 10 years compared to controls. In addition to documenting differences in the body composition of adolescent boys with sickle cell disease in Nigeria, this study also demonstrated the feasibility of using bioelectrical impedance to analyse the body composition of individuals under the hot, arid conditions which prevail in sub-Saharan Africa. PMID- 10822932 TI - Clinical features and treatment outcome of children with myeloid antigen coexpression in B-lineage acute lymphoblastic leukemia: a study of 151 Malaysian children. AB - The purpose of the study was to evaluate the incidence of myeloid antigen coexpression and its prognostic significance in childhood acute lymphoblastic leukemia (ALL) in Malaysia. A retrospective study was conducted of all ALL cases (< or = 12 years old) diagnosed and treated in University Hospital, Kuala Lumpur, Malaysia between 1 January 1992 and 30 May 1995, with available immunophenotype data. Presenting features and treatment outcome of 39 B-lineage ALL patients with myeloid antigen coexpression (My+B) were compared with 112 B-lineage ALL patients without myeloid antigen coexpression (My-B) for similarity in demographic, clinical and laboratory features and their treatment outcome. My+B and My-B patients were treated with a uniform treatment protocol. Myeloid antigen coexpression was defined as more than 30% isolated leukemic cells positive for CD13 and/or CD33. The ages at diagnoses ranged from 2 months to 12 years. Median age was 4 years. The incidence of myeloid antigen coexpression was 23 per cent. Univariate analyses showed that presenting features were similar between My+B and My-B with regard to age, sex, race, FAB morphology, white cell count, hemoglobin level, platelet count, liver/spleen size, central nervous system or mediastinal involvement, presence of lymphadenopathy, and proportion of blast cells detected in the marrow. Treatment outcome were not significant between the two groups. The 2-year event free survival was achieved in 44 per cent of My+B and 57 per cent of My-B (p = 0.11). The 2-year overall survival rates were 62 per cent for My+B vs. 77 per cent for My-B (p = 0.08). This study demonstrates that myeloid antigen coexpression is fairly common and constitutes 23 per cent of childhood ALL within the Malaysian population and that it is not an adverse risk factor in childhood ALL. PMID- 10822933 TI - Spontaneous and GnRH-provoked gonadotropin secretion and testosterone response to human chorionic gonadotropin in adolescent boys with thalassaemia major and delayed puberty. AB - To elucidate whether the cause of sexual maturation arrest in thalassaemia is of gonadal or pituitary etiology, 10 males with thalassaemia and delayed puberty and 10 with constitutional delay of growth and pubertal maturation (CSS) were extensively studied. Their spontaneous nocturnal gonadotropin secretion and gonadotropin response to intravenous 100 micrograms gonadotropin-releasing hormone (GnRH) were evaluated. Circulating testosterone concentration and clinical response were evaluated after 3 days, 4 weeks and 6 months of intramuscular administration of human chorionic gonadotropin (HCG) (2500 U/m2/dose). Thalassaemic boys had significantly lower circulating concentrations of testosterone compared to those with constitutional delay of growth and sexual maturation (CSS) at the same pubertal stage. Short- and long-term testosterone response to administrations of HCG was markedly decreased in thalassaemic boys. After 6 months of HCG administration 50 per cent (5/10) of the boys did not show significant testicular enlargement or genital changes. Despite the low circulating concentrations of testosterone, none of the patients had high basal or exaggerated gonadotropin response to gonadotropin releasing hormone (GnRH) stimulation. Luteinizing hormone (LH) peak responses to GnRH were significantly lower as compared to controls. Follicle-stimulating hormone (FSH) peak responses to GnRH did not differ among the two study groups. The mean nocturnal LH and FSH secretion was significantly decreased in all thalassaemic boys as compared to boys with CSS at the same pubertal stage (testicular volume). These data proved that hypogonadotropic hypogonadism is the main cause of delayed/failed puberty in adolescents with thalassaemia major. MRI studies revealed complete empty sella (n = 5), marked diminution of the pituitary size (n = 5), thinning of the pituitary stalk (n = 3) with its posterior displacement (n = 2), and evidence of iron deposition in the pituitary gland and midbrain (n = 8) in thalassaemic patients, denoting a high incidence of structural abnormalities (atrophy) of the pituitary gland. Moreover, in many of the thalassaemic boys, the defective testosterone response to long-term (6 months) HCG therapy denoted significant testicular atrophy and/or failure secondary to siderosis. It appears that testosterone replacement might be superior to HCG therapy in these patients. This therapy should be introduced at the proper time in these hypogonadal patients to induce their sexual development and to support their linear growth spurt and bone mineral accretion. PMID- 10822934 TI - Emergence of multidrug-resistant gram-negative organisms in a neonatal unit and the therapeutic implications. AB - Multidrug-resistant organisms are increasing worldwide. Over the years we have noted increasing resistance of organisms isolated in our neonatal unit. There is a need therefore to scrutinize the problem so as to be able to plan for the future. Over a 5-month period, 716 infants were admitted of which 192 were screened for sepsis. Overall, 121 (16.7 per cent) had positive blood cultures. The predominant organisms were Gram negative (73.6 per cent of isolates) with Klebsiella species topping the list at 31 per cent. Case fatality for infants infected with Gram negative organisms was 41 per cent. Resistance to gentamicin was 20 per cent chloramphenicol 23.6 per cent, and amoxicillin/ampicillin 66.3 per cent. Of worry is the resistance to ceftazidime 19.1 per cent, and cefuroxime 21.3 per cent, with the figures rising to 27 per cent when more specialized tests are done (disc approximation and potentiation tests). If these drugs cannot be used in 20-27 per cent of cases then the situation is serious. The contributory factors to increased resistance include: non-investigation of infants put on antibiotics (50 per cent of cases); prolonged (73 per cent) and sometimes unjustified (41.7 per cent) use of antibiotics; and non-utilization of investigations when these are done (52 per cent) together with the delay in getting results back in the ward (6 days). PMID- 10822935 TI - Inhibitory factors in breastmilk, maternal and infant sera against in vitro growth of Plasmodium falciparum malaria parasite. AB - Complications from falciparum malaria are responsible for over one million infant deaths annually. There is as yet no clinically protective vaccine that has been developed against human malaria parasites. While several studies have demonstrated the inhibitory properties of human sera against Plasmodium falciparum, there is no reported investigation that has examined the protective effects of human breastmilk against the malaria parasite. This study demonstrates the presence of significant antibody titers to ring, trophozoite, schizont and gametocyte stages of P. falciparum in 144 Nigerian maternal milk samples and also in paired maternal and infant sera. The study also demonstrates significant in vitro growth inhibition of P. falciparum by maternal and infant sera, but most notably by breastmilk samples and breastmilk constituents, such as lactoferrin and sIgA. The results therefore suggest a protective in vivo role for breastmilk in the possible modulation of malaria frequency, severity and complications. PMID- 10822936 TI - A new classification of acute protein-energy malnutrition. AB - Current schemes to classify protein-energy malnutrition (PEM) in hospitalized children below 5 years of age in developing countries are of limited usefulness, because age is often not known and wasted children are insufficiently discriminated. In this study, a newly proposed scheme, based on the presence of wasting (weight-for-height < or = -2 z scores) and oedema, was compared with clinical and the original Wellcome criteria to classify PEM in 538 under-fives hospitalized for PEM in a rural hospital in Nchelenge, Zambia. Wasting in the absence of oedema was clinically false-positively diagnosed in 34.0 per cent, whereas the Wellcome classification gave rise to 28.6 per cent false-positive and 39.9 per cent false-negative diagnosis. In the presence of oedema, clinical diagnosis of wasting was 44.7 per cent false-positive and 41.4 per cent false negative, while the Wellcome scheme had a false-negative rate of 54.3 per cent. We conclude that the Wellcome classification did not add to the clinical diagnostic accuracy of PEM as opposed to the proposed scheme, which merits further study. In an Appendix the use and calculation of z scores of weight-for height is further explained. PMID- 10822937 TI - Paediatric HIV infection in a rural South African district hospital. AB - The objective of this study was to determine the prevalence, clinical spectrum, and outcome of paediatric HIV infection in 281 consecutive children admitted to hospital in rural South Africa between October 1996 and January 1997. HIV infection was defined as two positive ELISAs in those aged > 12 months; a positive ELISA plus a positive IgG3 in those aged 6-12 months; and a positive ELISA plus positive p24 antigen or PCR in those aged 0-5 months. In all, 72 (26 per cent) children were HIV infected. Age-specific HIV prevalence was at least 25 per cent in all 1-5 year age groups. HIV-infected children were more likely to have been previously admitted (46 per cent vs. 23 per cent; p = 0.0002), and were more likely to have severe malnutrition (52 per cent vs. 17 per cent; p < 0.0001). Both HIV-infected and HIV-uninfected most frequently presented with diarrhoeal disease (51 per cent vs. 32 per cent), acute respiratory infection (13 per cent vs. 23 per cent), and malnutrition (18 per cent vs. 11 per cent). Satisfactory response to antibiotic therapy was less likely among the HIV infected (56 per cent vs. 73 per cent; p = 0.02), and mortality was higher among the HIV-infected (21 per cent vs. 7 per cent; p = 0.005). It is concluded that HIV-infected children present with disease syndromes common to this setting, but do so more frequently and with worse outcome than their uninfected counterparts. The high burden of paediatric HIV disease in this setting poses a substantial challenge for health resources. PMID- 10822938 TI - Human breastmilk storage and the glutathione content. AB - Human breastmilk storage for use later in infant feeding is on the increase as a result of the economic activities of nursing mothers. This study investigated glutathione (GSH) status of stored human breastmilk due to its major antioxidant role and as a cofactor for enzymes in detoxification of carcinogens. In newborns, human breastmilk becomes an important source of dietary GSH since their GSH synthetic capacity may not be well developed. The results showed that the total GSH content of human breastmilk obtained from apparently healthy lactating mothers was 192.2 +/- 148.3 mumol/l (mean +/- SD). Early breastmilk (fed to infants up to 4 weeks old; GSH content of 252.5 +/- 173.9 mumol/l) was significantly higher (p < 0.05) when compared with their mature counterpart (milk from mothers with infants older than 1 month of age; GSH content 163.9 +/- 128.0 mumol/l). Substantial loss of GSH occurred when breastmilk was kept at either -20 degrees C, 4 degrees C or at room temperature for 2 h. When compared with fresh unstored breastmilk, the extent of the loss was 80.6, 79.1 and 73.0 per cent respectively. It is suggested that feeding infants on stored human milk could weaken the antioxidant and toxin refractory capacity of those in early childhood. PMID- 10822939 TI - Total anomalous pulmonary venous drainage: mixed type. PMID- 10822940 TI - Improved pediatric weighing device for use with portable hanging scales. AB - Portable hanging scales are a common means of weighing young children. Weighing trousers used to suspend children from a hanging scale are uncomfortable and require insertion of the child's legs through the trousers. The 'weighing seat' holds the child in a more natural, comfortable, and secure position, thus increasing the child's acceptance of the weighing procedure, and making the procedure less traumatic for the child and easier for the examiner. PMID- 10822941 TI - Children with cystic fibrosis in South Africa: an improving nutritional picture. AB - Nutritional status and growth play an important part in determining the prognosis in cystic fibrosis (CF). In South Africa, the median survival of patients with CF is 18 years. Using chart review, we studied the pattern of growth over time of a South African CF population. The percentages of expected weight-for-age, height for-age and weight-for-height were determined for each patient in 1986 (n = 49) and 1996 (n = 63). Mean indices were the same in the two years. In 1996, mean weight-for-age of children aged 5-10 years was 94.2 per cent (SD 20.4), 14.3 per cent higher (p < 0.05, 95 per cent confidence intervals 3-25 per cent) than children of the same age in 1986. Improved growth of young children with CF has been achieved in a resource-poor country setting the scene for improved prognosis. PMID- 10822942 TI - Secular trends of HBeAg prevalence among HBsAg-positive delivery mothers in a hepatitis B endemic area. AB - A high prevalence of HBeAg among HBsAg-positive mothers at the time of delivery results in a high prevalence of hepatitis B vertical transmission. From 1990 to 1995, 896 pregnant HBsAg-positive women, including 411 (46 per cent) HBeAg positive subjects, were enrolled in our study to analyse the secular change in HBeAg prevalence. Their mean age, number of pregnancies and parity were 29.5 +/- 4.1 years, 2.0 +/- 1.2, and 0.6 +/- 0.7, respectively. The prevalence rates of HBeAg were 48, 54, 49, 47, 40, and 40 per cent among the subjects enrolled in 1990, 1991, 1992, 1993, 1994, and 1995, respectively. In univariate analyses, prevalence of HBeAg decreased by the calendar year of pregnancy (p = 0.01), and also by age (p < 0.00001), number of pregnancies (p < 0.0001) and parity (p < 0.0002). After adjusting for age in multiple logistic regression, the calendar year of pregnancy was still the independent variable, while gravida and parity became insignificant. The odd ratios (95 per cent confidence interval) of HBeAg negative-seroconversion in the equations were 1.09 (1.00-1.19) per calendar year and 1.14 (1.10-1.18) per year of age. Our results have shown a secular decrease in HBeAg-prevalence among pregnant HBsAg-positive women in Taiwan. PMID- 10822943 TI - Diagnostic value of bone mineral density measurements in infants with rickets. PMID- 10822944 TI - [Bernardino Ramazzini and "Medicina del Lavoro"]. AB - Three hundred years after the first edition of Ramazzini's "De Morbis Artificum Diatriba", this paper reviews all the articles on the work of the great Master of Carpi that have been published over a century in the ninety volumes of "La Medicina del Lavoro". Since the first issue of the journal in 1901, many authors have submitted papers which revisited and commented on the work of Ramazzini, particularly in anniversary years (1914, the anniversary of his death, 1933 and 1983, anniversaries of his birth), but only Luigi Devoto was capable of fully understanding the methodology introduced by Ramazzini, transforming it into practical applications which he listed in ten fundamental actions. Mainly for this reason and considering that Ramazzini had been practically ignored for two centuries, it was decided to consider the Devoto period as the "true Ramazzini era". The death of the founder of the journal in 1935 marked the unexpected beginning of a long silence, lasting up to 1983, during which not one paper on Ramazzini was published in "La Medicina del Lavoro". It is concluded that a comparison between the diseases described by Ramazzini and those of to-day is of no practical use and that a tribute to Ramazzini is simply necessary in recognition of his contribution to the birth and development of occupational medicine, but is nonetheless insufficient. What is also needed, following in the footsteps of the founder of this journal and of the Clinica del Lavoro of Milan, is to reinterpret Ramazzini's methods in modern terms in the perspective of the ten actions he proposed, bearing in mind that possibilities exist to-day whereby his efforts can be completed and further enlarged in terms of practical applications. PMID- 10822945 TI - [Tumor and work: 20 years after "The causes of cancer" by Doll and Peto]. AB - In the last twenty years scientific research in the field of occupational cancer has achieved some outstanding results. The number of substances and work processes classified as carcinogens for man has doubled. New cancer sites have been associated with exposure to known occupational carcinogens. New occupational risk factors for cancer sites already known as targets for occupational exposures have been identified. The agents responsible for the excess cancer risk in some industrial settings have also been identified, as well as the mechanisms by which numerous occupational risk factors play a role in the various steps of the multifactorial carcinogenic process. Experimental studies have provided strong support for this progress but substantial contributions have been made as a result of the increase in the number of studies of occupational cancer epidemiology, the establishment and use of large data-bases, the increasing tendency to planning of multicentric epidemiological studies, and the constant improvement of retrospective exposure assessment methods with the aid of more sophisticated job-exposure matrices and the use of biomarkers, some deriving from the old biological monitoring programs, others only recently introduced. However, in spite of the attention that recent Italian legislation for the prevention of occupational diseases has devoted to the problem of occupational cancer, a gap between scientific research in the field of occupational cancer and the practice of occupational health persists in Italy. Future studies can help to close this gap by integrating multidisciplinary contributions from clinicians, toxicologists, epidemiologists, and industrial hygienists right from the planning stage. PMID- 10822946 TI - [Prevalence of carpal tunnel syndrome in workers executing complex tasks with the upper extremities]. AB - In 50 female workers of a manufacturing plant employed on the assembly line of metal and plastic products with cloth covering, we found a high prevalence of Carpal Tunnel Syndrome (16 cases, 32%, 5 of whom had already undergone surgical treatment). The subjects with positive diagnosis of Carpal Tunnel Syndrome also had a longer occupational exposure to repetitive movements, compared to the other subjects. Among the subjects with this disorder, the length of exposure was also significantly associated with the reported symptoms and the extension threshold measured on the 2nd finger of the dominant hand. Ergonomic assessment of the assembly lines revealed high-frequency repetitive movements and application of force, with insufficient rest intervals. PMID- 10822947 TI - [Mortality in employees of a railway rolling stock factory]. AB - A study was carried out on the mortality of 3741 individuals who worked in a factory making railway rolling stock, in which crocidolite was used to insulate the coaches in the 60's and 70's. Employees who had worked in the factory between 1960 and 1995 were included in the study, with the main aim of investigating cancer mortality in these subjects. 9 cases of mesothelioma were known in this factory before starting the study. The follow-up was performed from 1960 to 1996. The SMR's for lung cancer and mesothelioma in 2737 blue-collar workers were 1.23 (CI 95% = 1.00-1.50) and 6.50 (CI 95% = 2.96-12.38) respectively, based on national rates. The excesses were mainly found among subjects who had worked before 1970 with duration of employment exceeding 20 years. PMID- 10822948 TI - [Measurement of airborne asbestos fibers on railroad rolling stock]. AB - In February 1995 the Italian Railways Health Department set up a special study group in order to assess the effectiveness of the measures adopted against hazards due to the presence of asbestos in rolling stock currently in use on the rail network. The group set up specific procedures for sampling and analysis, on the basis of the criteria fixed for civil buildings in Ministerial Decree of 6/9/94, which was subsequently applied to rolling stock by Ministerial Decree of 26/10/95. In accordance with these procedures the study group carried out environmental studies via test runs programmed by the Railways Technical Departments, on trains made up of different types of vehicles. Insulated, completely or partially deinsulated and originally non-insulated vehicles were studied. Samples were analysed via scanning electron microscopy (SEM) with elementary dispersion X spectroscopy (EDXS) carried out by highly qualified public laboratories (ISPESL--National Institute for Prevention and Work Safety, ARPA--Regional Environmental Protection Agency, CRA--Veneto Region, University Departments). Altogether, from the start of the programme up to September 1998, 1464 samples in 170 test runs on 619 rolling stock vehicles were examined. These involved 83 locomotives, 83 electric rail-cars and 453 carriages. The results showed that in over 99% of the samples the fibre concentrations were below 2 fibres/litre, which is the value fixed by law for buildings and rail vehicles in order to qualify for effective decontamination status. Values exceeding 2 fibres/litre were found in only 4 vehicles, which were withdrawn or blocked for further checks. As a precaution, 18 vehicles where concentrations over 1 but less than 2 fibres/litre were found, were also blocked and their return to service has been postponed for further checks and analyses until the results show concentrations below 1 fibre/litre. Environmental analyses carried out up to the present indicate an overall situation comparable to that usually found in the general environmental without any asbestos dispersion sources. Surveillance and investigations are still under way in order to achieve the aims of the safety programme set up by the Italian Railways to ensure health and environmental protection. PMID- 10822949 TI - [Follow-up of allergic symptoms in a group of health workers sensitized to latex]. AB - We studied 28 health care workers sensitised to latex and complaining of respiratory symptoms at work. All were females, aged 32 y (median), and had worked in the hospital for 12 y, in departments with high exposures (22 cases), moderate exposure (4 cases) or low exposure (2 cases) to rubber gloves. Twenty also complained of latex skin symptoms. Twenty-five (89%) were atopical (by prick test), and 16 (57%) had a personal history of allergic disease. Nine (32%) also had symptoms from exposure to latex outside work and 3 (10%) from food cross reacting with latex. The specific inhalation test with latex gloves confirmed the diagnosis of asthma in 4 cases (14%) and of rhinitis in 13 cases (46%). We reinvestigated the workers 14 months (median) after the first examination to update exposure to latex, any allergic symptoms and adverse effects of relocation. At the follow-up, 3 workers (11%) had retired, all the others had stopped wearing powdered latex gloves, and 12 had changed their duties or the work-place. Ten (40%) of the 25 subjects still at work were without symptoms, 14 (56%) occasionally complained of mild symptoms and one of frequent rhinitis. Overall, we observed improvement of skin symptoms in 74% (14/19) of the cases and of respiratory symptoms in 56% (14/25) of the cases. Symptoms from exposure to latex outside work were still present in 6 subjects. The persistence of symptoms at work was significantly higher among workers who continued to work in departments with high/moderate latex glove exposure. The results of the study show that skin and respiratory symptoms among health care workers are related to the use of powdered latex gloves and confirm the importance of primary prevention. PMID- 10822950 TI - [Organization of health and safety activities in health care centers]. AB - The paper proposes a model for health and safety organization in health care units and hospitals which takes account of the risk assessment procedures required by law and the quality assessment of the measures thus taken. A redefinition is given of the role of Medical Director and of the functions, aims and standards on which health and safety service and the services of an authorized occupational health physicians must be based. PMID- 10822951 TI - [Use of experimental data in percutaneous absorption in the process of risk assessment. Dermal Exposure Network Percutaneous Penetration Subgroup]. PMID- 10822952 TI - [Difficult retirement from the white coat. Does great emptiness follow general practice role?]. PMID- 10822953 TI - [Increased refusal of natural childbirth among women. Cesarean section by request?. Interview by Dr. Brigitte Moreano]. PMID- 10822954 TI - [Noise effects not only the ears. But can damage to health be objectively evaluated?]. AB - Noise is an unwanted sound and/or a potential health hazard. Apart from noise induced hearing loss noise causes numerous psychosocial and unspecific vegetative effects. Disturbances of communication are most frequently complained, and sleep disturbances ar regarded as most deleterious. These reactions may then cause performance decrements, annoyance and behavioral alterations. The permanent influence of noise may contribute to the multifactorial genesis of cardiovascular diseases, particularly of hypertension. PMID- 10822955 TI - [Arm paralysis after motorcycle accident. When the surgeon should be consulted in brachial plexus lesions]. PMID- 10822956 TI - [A simple aid in gastroenterology. Even the stomach tube has its risks]. PMID- 10822957 TI - [Chronic cough with muscle stiffness. Bronchial carcinoma]. PMID- 10822958 TI - [Risks to the subsistence of the internist!]. PMID- 10822959 TI - Births, marriages, divorces, and deaths: provisional data for May 1999. PMID- 10822960 TI - [From the standpoint of medical ethics]. PMID- 10822961 TI - Craniofacial surgery for malignant tumors of the paranasal sinuses. PMID- 10822962 TI - [Results from the treatment of nasopharyngeal cancer]. AB - Nasopharyngeal carcinoma constitute 0.3% all malignancies in Poland. These tumors still pose a complex diagnostic problem. Intracranial structures and facial skeleton have not been accessible for X-ray imaging so far. Improvement o imaging techniques, fibroscopy, computed tomography, magnetic resonance imaging allows for gathering new data which influence diagnostics and treatment planning in this difficult anatomical localization. Development of radiotherapy techniques allows for decreasing of radio-reaction and neighbouring structure damage. 188 patient were included in this series and divided into two subgroups: I--128 patients treated between 1980-1990 and II--60 patients between 1991-1995. The diagnostics and treatment methods differs in both subgroups. It was connected with new imaging techniques, therapeutic equipment as well as combination of radiotherapy and neoadjuvant chemotherapy. The authors present the latest diagnostic and therapeutic methods which are currently applied in ENT Dept. Karol Marcinkowski University of Medical Sciences and Wielkopolskie Centrum Onkologii with review of literature devoted to the progress in diagnostics, treatment planning and modern radiotherapy techniques. PMID- 10822963 TI - [The course of oropharyngeal stage of deglutition after partial supracricoid laryngectomies. Computerized analysis of radiographic imaging. Preliminary report]. AB - On a pilot basis the performed analysis covered the pharyngeal phase of the deglutition act in five patients, in whom supracricoid laryngectomies were carried out due to carcinoma of the larynx. The rentgenocinematographic (Rtgc) examinations were made half a year after the operations, while in 1 patient the studies were carried out twice, the second being performed two years after the operative procedure thus altogether 6 examinations Rtgc were performed. Computerized topocinetic analysis of images Rtgc was made intended for further considerations and measurements, and concurrently assessing the position of the left-behind anatomical structures, the pathway passed by the studied structures, the velocity, the projection of the encircled kinetic path, as well as the distance of individual structures in respect to each other during a complete cycle of the pharyngeal phase. The provided description highlights the significance of good mobility of the left structures in the food bolus passage trough the lower pharynx and other, than that after supraglottal laryngectomy, mechanism protecting the lower respiratory tracts against food leakage into them. PMID- 10822964 TI - [Free forearm skin flaps with microvascular anastomosis for pharyngeal reconstruction]. AB - Free forearm skin flap with microvascular anastomosis was used for tissue defect reconstruction in 4 patients after resection of the oropharynx, base of the tongue and ramus of the mandible. The vascular pedicle of the flap contained the radial artery and the cephalic vein. For microvascular anastomosis the facial artery was used in all the patients, the facial vein in 2 patients, the internal jugular vein in one patient and the external jugular vein in the other one. Good healing of the graft was obtained in 3 patients. In one patient partial necrosis developed. No patient had fistula. PMID- 10822965 TI - [Tonsillectomy as a routine treatment in cases of branchiogenic cancer]. AB - The pathogenesis of the branchiogenic cancer is still unknown. A number of authors think that it is a metastasis in the form of a cyst from a primary focus in the pharynx most often located in the palatal tonsil. Hence a conclusion is put forward to perform a tonsillectomy even when a tonsil is not clinically changed. The author presents a medical record of a patient with plano-epithelial carcinoma of a cyst from a primary focus in the pharynx most often located in the palatal tonsil. Hence a conclusion is put forward to perform a tonsillectomy even when a tonsil is not clinically changed. The author presents a medical record of a patient with plano-epithelial carcinoma of a lateral cervical cyst. It developed in the epithelium, sub-epithelium and in the palatal tonsil. The tonsillectomy was performed in spite of the clinically unchanged. This case indicates that cancer can develop simultaneously in the cyst wall and in the tonsil. The presence of the wall architecture elements typical of a lateral cervical cyst and the duct connecting the cyst with the wall of the pharynx indicates that it wasn't a metastasis from the focus in tha tonsil. The development of both these focuses could have resulted from the spreading of a lesion through continuity in the ascending way (a cyst-a tonsil) or through the descending way (a tonsil-a cyst). The author suggests that a branchiogenic cancer is an indication for a routine prophylactic ipsilateral tonsillectomy and neck dissection together with consecutive irradiation. PMID- 10822967 TI - [Second primary malignant tumor in patients with nasopharyngeal cancer]. AB - Clinical analysis of 90 patients with malignant nasopharyngeal tumors treated in 1985-1996 was performed. The real frequency of second tumors, immunity to second malignant tumor, therapy and genetical factors were described. Second malignant nasopharyngeal tumors occur rarely than in other head and neck tumors (in our material approx. 4%). An attention should be paid to the young age of our patients, mean 46 years. PMID- 10822966 TI - [The use of microsurgery and balneotherapy in the treatment of hyperplastic laryngitis]. AB - The study aimed at the assessment of the clinical value of using the combined method of laryngeal microsurgery and balneotherapy in the operative treatment and prevention of recurrences of limited hyperplastic changes of the laryngeal mucosa. The study material comprised two groups of patients diagnosed and treated in the years 1991-1997 in the Lublin E.N.T. Department and Laryngological Research-Consulting Center in Iwonicz Zdroj. The I clinical group consisted of 194 patients in whom, on definite indications, micro-surgeries with Kleinsasser's method were performed and the II control group consisting of 165 subjects suffering from chronic simple inflammation of pharyngo-laryngeal mucosa without morphologic hypertrophy. Inhalant treatment was performed by means of an individual method--using inhalant devices by Thomex L-2 producing high density mist of 1-4 mu degree comminution. Isotonic Iwonicz saline-Elin 7 was used. Uniform criteria of assessment were assumed and used of direct and distant therapeutic effect (in control studies) for the whole group of patients in the study period. Obtained results were analysed statistically. PMID- 10822968 TI - [Pharyngeal closure after pharyngolaryngectomy]. AB - Pharyngolaryngectomy was performed in 53 patients. In 36 patients pharynx defect was less than 50% of pharynx circumference and it was closed without reconstruction. Larger pharynx defects were closed using platysma myocutaneous flap (13 cases), pectoralis mayor flap (3 cases) and free forearm flap with microvascular anastomosis (1 case). Healing results are presented in each group of patients. PMID- 10822969 TI - [Monitoring of the inflammation in children before and after tonsillectomy]. AB - A group of 54 children aged from three to 13 years was qualified to tonsillectomy for laryngological indications. In sera of all children following measurements were performed: the concentrations of C-reactive protein (CRP), alpha1-acid glycoprotein (AGP) and alpha1-antichymotrypsin (ACT) were measured using rocket immunoelectrophoresis according to Laurell, also concentrations of three main immunoglobulin classes (IgA, IgG, IgM) and antistreptolysin titer. The microheterogeneity of both AGP and ACT was investigated, using crossed affinity immunoelectrophoresis according to Bog-Hansen with Concanavalin A (Con A) as a ligand. Results were expressed as reactivity coefficients (RC), being the proportion of all Con A-reacting variants to the non-reacting variant. It is worth mentioning that there was no difference in all investigated parameters as well between groups obtained by categorizing children according to the presence or absence of elevated antistreptolysin titer. It may mean that at least in some cases the chronic inflammation was caused by streptococci non-producing streptolysin O. The results obtained suggest that the absence of the arthritic pain does not exclude the need of antibiotic therapy in children after tonsillectomy. Generally it is postulated that estimation of acute phase proteins concentrations and glycosylation profiles, which were previously shown to be useful in clinical assessment of various diseases may serve as additional marker in laryngology in cases where indications to tonsillectomy are still controversial. PMID- 10822970 TI - [Diagnostic difficulties in nasopharyngeal tumors]. AB - Diagnostic possibilities in lethal midline granuloma (Wegener's granulomatosis, angiocentric lymphoma) based on the literature were presented. Results of diagnostic examinations in a case of 34 years old man admitted to the Dept. of Otolaryngology in Poznan were described. PMID- 10822971 TI - [Transient evoked otoacoustic emissions (TEOAE) in tinnitus patients treated with xylocaine]. AB - The group of patients suffering from tinnitus was treated with intravenous Xylocaine. Ten consecutive doses of Xylocaine (2 mg/kg b.m.) were administered during a period of ten days. Changes of amplitudes of TEOAE in relation to tinnitus suppression was assessed. In 91.5% of cases we observed correlation between amplitude changes and complaint abatement. Both amplitude changes and the time of tinnitus suppression increased after ten days of treatment in comparison with single dose of Xylocaine. The differences of amplitude variations between tinnitus and non tinnitus ear group was statistically significant (Wilcoxon test p < 0.05). The treatment with Xylocaine was considered to be the beginning of long-term procedure aimed at causing habituation of tinnitus in patients conscience (TRT). PMID- 10822972 TI - [Voice disorders in children with gastroesophageal reflux disease]. AB - The study concerns 11 patients with gastroesophageal disease confirmed by pH metric examination of esophagus. The clinical assessment included lupolaryngoscopy, stroboscopy and acoustic analysis of voice. Basing on the tests there were confirmed inflammatory lesions at the posterior part of larynx (laryngitis posterior) as well as pathological changes of voice acoustic tests. PMID- 10822973 TI - [Videolaryngostroboscopy of the larynx in patients after thyroid surgery]. AB - The authors have presented the video of different stroboscopic pictures of the larynx in patients after thyroid gland surgery. PMID- 10822974 TI - [Central transmission time in children with the delayed language development]. AB - The prolonged central transmission time (CTT) in ABR examinations in children with the delayed speech was described in 1992. Now after 4 years the examinations were repeated. The children with actually good speech revealed normalization of CTT, but in children with dyslalia and dyslexia this interpeak latencies were once again prolonged. PMID- 10822975 TI - [Desmoid tumor of parotid gland]. AB - Desmoids are rare tumors originating from musculo-aponeurotic soft tissues. Their biologic features placed them between benign fibromas and malignant fibrosarcomas. The rare of parotid gland desmoid tumor is presented. The patient was treated surgically--the tumor was resected with the safety margins. Recurrence was not observed during 12 month follow up. PMID- 10822976 TI - [The development of concept of partial surgery of the larynx due to supraglottic cancer]. AB - It has been shown the conception of partial surgery of the larynx due to cancer primary develops in the supraglottic. The evolution of this method which is a consequence of progress in anatomical and embryological knowledge as well as technical possibilities of surgeons has been introduced. The modern methods of partial removal of larynx let resume physiological function of larynx with oncological security. PMID- 10822977 TI - [Professional education of Polish physicians in otologic centers of Vienna in the 19th century]. AB - The foundations of the modern otology in Vienna created Adam Politzer, Joseph Gruber, Victor Urbantschitsch. The professional and scientific achievements of Joseph Gruber and Victor Urbantschitsch are described briefly. Gruber was one of two heads of otiatric clinic in Vienna. Urbantschitsch was an authority of psychophysiology of hearing. Such well-known otologists in Vienna as Gustav Alexander, Robert Barany, Beniamin Gomperz, Erich Ruttin and others were a good teachers of many foreign physicians also from Polish territories. Robert Barany was a Nobel prize winner. PMID- 10822978 TI - [Mould hypersensitivity in patients suffering from allergic rhinitis]. PMID- 10822979 TI - [Multifrequency tympanometry in the diagnosis of acoustic organs]. PMID- 10822980 TI - Biological indicator comparative analysis in various product formulations and closure sites. PMID- 10822981 TI - Isolators--validation and sound scientific judgement. PMID- 10822982 TI - Airborne viable particles and total number of airborne particles: comparative studies of active air sampling. AB - This paper presents results from a comparative study of different impaction air samplers under simulated operational conditions in a controlled environment. Furthermore measurements have been performed in a body-box during evaluation of clean-room clothing and in a day care center. Microbial monitoring of controlled environments can be performed in several ways, and with the aid of different sampling instruments. Knowing the limitations of the chosen system is of vital importance for the correct evaluation and interpretation of the results. The number of Colony Forming Units (CFU) detected by one method cannot be directly compared with results from another method. The obtained results seem to indicate that in a controlled environment the exposure situation with a human contamination source gives an approximate relationship between the total number of airborne particles and the number of aerobic airborne viable particles detectable with standard methods. PMID- 10822983 TI - Carrying out biological qualification, the control operation of moist-heat (steam sterilization) processes for producing sterile pharmaceuticals and medical devices. AB - In this report we will first discuss the principles behind the practices that are used today in the design and qualification of moist-heat (steam sterilization) microbial-control processes used to produce sterile pharmaceutical products and medical devices. Secondly, we will work through example applications of how to design and qualify processes of three levels of complexity which we call Empirical Overkill, Empirical, and Product Specific. Empirical Overkill is specifically for the microbial-control processes for indirect items, such as tanks, pipes, pumps and other hardware. Empirical is for pharmaceutical and medical-device products that are produced under good manufacturing conditions and, therefore, there is control of the level of the microbial bioburden. Product Specific is for microbial-control processes designed and qualified for a specific product on the basis of the numbers and resistance of the bioburden of that product. We will treat design in this report; however, the major thrust is in setting up and carrying out the biological qualification of the process, which is the mode of control used to assure the adequacy of these microbial-control processes. PMID- 10822984 TI - Photodestabilization of epinephrine by sodium metabisulfite. AB - The effect of light and bisulfite on infusion solutions containing epinephrine (2 micrograms/mL), bupivacaine hydrochloride (1 mg/mL), fentanyl (2 micrograms/mL) and sodium edetate (0.18 microgram/mL) was investigated. Solutions were made isotonic using sodium chloride, and they contained either 1.82 micrograms/mL sodium metabisulfite or no metabisulfite at all. The infusions were stored in polypropylene infusion bags (550 mL) and were subjected to light from a xenon burner with a specter similar to sunlight behind a glass window. The contents of epinephrine, bupivacaine and fentanyl were determined with stability indicating methods using a HPLC system. In two identical experiments epinephrine was demonstrated to be more stable under irradiation in the absence of sodium metabisulfite (89.3% and 91.3% remaining of initial concentration) than in its presence (64.5% and 60.1% remaining). Bupivacaine and fentanyl showed no degradation as a function of light exposure, and the stability was not affected by the presence or absence of bisulfite. After two weeks exposure to natural light (Oslo, 60 degrees N, March) the content of epinephrine was 45.0% of the initial concentration in the solutions containing bisulfite and 95.6% in the solutions without bisulfite (n = 3). The relationship between various concentrations of bisulfite and the photo-stability of three epinephrine solutions (2 micrograms/mL, 20 micrograms/mL and 110 micrograms/mL) were tested. The results indicated that approximately equimolar concentration of epinephrine compared to bisulfite, or bisulfite in some excess, was least favorable to the stability of epinephrine. A possible explanation for the photo-destabilizing effect of bisulfite can be the conversion of superoxide radicals (O2-) to highly reactive hydroxyl radicals (.OH) by bisulfite. From these results it can be concluded that solutions containing epinephrine and bisulfite should be protected from light not only during storage but also during use in cases where the infusion time amounts to several days. An alternative, wherever possible, would be to omit bisulfite in the solutions. PMID- 10822985 TI - Moisture sorption behavior of selected bulking agents used in lyophilized products. AB - To develop a rational approach for the formulation of lyophilized products, six bulking agents commonly used in freeze-dried formulations were lyophilized under identical conditions, and their moisture sorption behavior, before and after lyophilization, were determined as a function of relative humidity at 25 degrees C. The bulking agents evaluated were mannitol, anhydrous lactose, sucrose, D(+) trehalose, dextran 40 and povidone (PVP K24). The materials were also characterized for their crystal and thermal properties by powder X-ray diffraction, DSC and TG after exposure to various relative humidity conditions. Mannitol was crystalline and non-hygroscopic both before and after lyophilization with total moisture contents of 0.1 to 0.3% w/w between 10 and 60% RH. Anhydrous lactose, sucrose and trehalose were crystalline prior to lyophilization with moisture contents of 0.86, 0.15 and 9.2%, respectively, and the crystalline materials were relatively non-hygroscopic. Upon lyophilization, they converted to the amorphous form and had moisture contents of 1.6, 2.5 and 1.2%, respectively. The amorphous materials sorbed moisture rapidly upon exposure to increasing relative humidity conditions. The amorphous lactose converted to its crystalline hydrate form at 55% RH after sorption of an additional 10% moisture. This conversion to the crystalline hydrate form was accompanied by desorption of practically all the moisture sorbed by the amorphous form. Similarly, lyophilized sucrose converted to its crystalline form after the sorption of additional 4.5% moisture at 50% RH, and the lyophilized trehalose sorbed additional 10% moisture prior to its conversion to a crystalline hydrate form at 50% RH. Dextran and povidone were amorphous and hygroscopic both before and after lyophilization and they sorbed as much as 10-20% moisture at 50% RH. It is well established that different drugs, especially proteins, need different levels of moisture for optimal stability. The results of the present study show that moisture contents of lyophilized cakes may be varied and optimized by the selection of suitable excipients. PMID- 10822986 TI - Opinion: Kajander's nanobacteria. PMID- 10822987 TI - Parenteral formulations of small molecules therapeutics marketed in the United States (1999). Part III. PMID- 10822988 TI - [History of Krakow hematology]. AB - The development of the studies of the blood diseases at the IIIrd Department of Internal Medicine and later at the Department of Hematology of the Medical Academy and subsequently the Collegium Medicum at the Jagiellonian University in the years 1950 to 2000 are presented here. The Cracow Hematological Center, headed by professor Julian Aleksandrowicz from 1950 to 1979, by professor Julian Blicharski from 1979 to 1989, by ass. professor Jerzy Lisiewicz from 1990 to 1993, and presently by Aleksander B. Skotnicki--has drawn many brilliant hematologists who have initiated, developed and verified new concepts of pathogenesis as well as new methods of diagnosis and therapy of haematological diseases. The Polish Haematological Society was founded in 1949 in Cracow where the first Conference took place in May 1950. The first specialistic haematological journal--Haematologica Cracoviensia later retitled as Haematologica Polonica was first published in Cracow in 1957. Finally here, in Cracow, one of the first syngeneic bone marrow transplantation was performed in 1958. Presently the Chair and Department of Haematology of the Collegium Medicum at the Jagiellonian University is a modern and well facilitated center of research and didactics (both pre and post graduate studies); the modern cytological, immunophenotype, cytogenetic and molecular diagnostic methods are used in haematological patients from the Macroregion of South-Eastern Poland (inhabited by approximately 8 million people). The offered treatment includes chemo- and radiotherapy, immunotherapy and autologous and allogeneic bone marrow transplantation. PMID- 10822989 TI - Bone marrow and haematopoietic stem cell transplantation in Poland. AB - The concise history of BMT development, from experimental studies to clinical practice, is presented. Bone marrow transplantation has been performed in Poland for more than ten years, starting with a few to almost 500 procedures up to 1999. The above was accomplished thanks to successive founding of the 14 active BMT centers throughout Poland, which presently perform from 10 to 130 BMTs per annum (mean 34). About 70% of the above number are autologous BMTs, the remaining are allogeneic BMTs from HLA compatible sibling donors and recently from compatible unrelated donors (URD). During the last few years there is an increasing trend in the number of hematopoietic stem cell transplantations derived from peripheral blood (PBSCT), not only in patients with leukaemias and malignant lymphomas, but also in patients with solid tumors. The BMT results achieved in Poland correspond to results obtained worldwide. In relation to other European countries there are less BMT teams (about 50%) and less BMT procedures (also about 50%) performed per annum (according to EBMT data calculated per 10 million inhabitants). Thanks to the financial support of the Polish Ministry of Health more and more patients are qualified for high dose chemotherapy, increasing their chances for long term disease free survival and, eventually, for cure. PMID- 10822990 TI - Advances in treatment of chronic myeloid leukaemia. PMID- 10822991 TI - The role of methylation in CML. AB - Methylation of the proximal promoter of the ABL1 oncogene is common epigenetic alteration associated with clinical progression of chronic myeloid leukemia (CML). In presented study we queried whether both the Ph'-associated and normal ABL1 alleles undergo methylation; what may be the proportion of hematopoietic progenitors bearing methylated ABL1 promoters in chronic versus acute phase disease; whether methylation affects the promoter uniformly or in patches with discrete clinical relevance; and, finally whether methylation of ABL1 reflects a generalized process or is gene-specific. To address these issues, the technique of methylation-specific PCR and bisulfite-sequencing was adapted to study the regulatory regions of ABL1 and other genes. In cell lines established from CML blast crisis, which only carry a single ABL1 allele nested within the BCR-ABL fusion gene, ABL1 promoters were universally methylated. In clinical samples from patients at advanced stages of the disease, both methylated and unmethylated promoter alleles were detectable. In colonies derived from single hematopoietic progenitors methylated and unmethylated promoter alleles were revealed as well. ABL1 methylation was was noted in the vast majority of colonies from blast crisis, but not chronic-phase CML. It was shown finally that ABL1 methylation does not reflect a generalized process and may be unique among DNA repair/genotoxic stress response genes. These data suggest that specific methylation of the Ph'-associated ABL1 allele accompanies clonal evolution in CML. PMID- 10822992 TI - Experience with the first 200 allogeneic blood cell transplants (BCT) in Calgary. AB - Allogenic peripheral blood stem cells transplantation (BCT, allo-PBSCT) have an established place in currently active protocols at the University of Calgary, we now use BCT as the third approach for acute myeloid leukemia (AML) in first complete remission (CR). For chronic myelogenous leukemia (CML) beyond first chronic phase (CP) and other hematological malignancies BCT gives outcomes at least equivalent to bone marrow transplantation (BMT). BCT recipients benefit from faster engraftment. However a tendency to develop more graft-versus-host disease (GVHD) is observed. Bone marrow is still the preferred source of stem cells for CML in first CP and for patients with non-malignant disorders. The ability to obtain high number of blood cells allows us to be able to perform haptoidentical transplants using T-cell depletion and to obtain engraftment in most cases. Refining tha latter technique however remains a challenge particularly with respect to immune reconstitution. PMID- 10822993 TI - Current status of high dose chemotherapy in breast cancer. AB - To date, no definite answers are available to the initial question, whether or not HDCT and ASCT can improve quantity or quality of life. Completing the available prospective, randomized phase III studies; using HDCT earlier in the course of treatment; applying drugs that are active breast cancer; comparing the experimental treatment to standard therapy; and using appropriate sample size to detect clinically meaningful and statistically significant differences are all paramount requirements to answer the question successfully. Within the next few years, a number of European [13] and two Canadian Studies [6, 14] that are currently accruing patients, will contribute versus important information regarding the role of HDCT and ABMT for breast cancer. PMID- 10822994 TI - Flow cytometric assessment of bone marrow transplant patient. AB - Flow cytometry laboratory plays an integral part in the evaluation of a BMT patient. Its role starts at the time of clinical presentation to assure most accurate and reproducible disease diagnosis and subclassification. This can only be achieved if the flow cytometrist is fully qualified in the diagnosis of hematolymphoid neoplasia and is able to correlate each and every case with morphologic data. Flow cytometric report, therefore, includes a standing diagnosis and goes beyond the sole description of an abnormal/atypical population. Following initial diagnosis, generated data are carefully stored and are retrieved for review whenever needed particularly during the evaluation of the remission status, response to chemotherapy, relapse, staging and detection of residual disease in marrow and apheresis product. Our laboratory performs leukemia/lymphoma assessment of approximately 150-200 samples a month in addition to 3-8 weekly apheresis procedures, 25-30 post-BMT immune monitoring samples and other isoteric assessments including CD4 counts for HIV population, HLA B27 evaluation, platelet/leukocyte antibodies, reticulated platelets, immunodeficiency disorders etc. This multidisciplinary center allowed us to develop a substantial expertise in the field, which hopefully benefits our patient, clinicians and fellow cytometrists. We are please to share our expertise at this distinguished forum. PMID- 10822995 TI - Bone marrow examination for primary diagnosis and follow-up in patients with lymphoproliferative disorders. An overview with emphasize on bone marrow histology. AB - The aim of his overview is to provide some insight into the manifold applications of bone marrow examination in lymphoproliferative disease. Having a large selection of methods any clinician working in this field should be familiar with the diagnostic relevance of each method for a given diagnostic problem. The choice of the appropriate investigations will warrant a correct diagnosis and helps to lower health expenses. Because of the multiplicity of the methods, often performed at different institutions, a lot of confusion can be caused by conflicting interpretation of isolated results. A diagnosis should never be based on isolated findings such as immunophenotyping or demonstration of a clonal population. Results of various investigations are to be integrated into a final diagnostic concept, which will be the basis for therapeutic decisions. PMID- 10822996 TI - Bone marrow pathology after bone marrow transplantation. AB - Trephine biopsy evaluation in patients who have undergone high dose chemotherapy with stem cell infusion performed by an experienced pathologist is important in evaluation of hematopoietical restitution, evaluation of complications such as graft rejection, infections, relapse or secondary neoplasms. In the first week after high dose chemotherapy or radiotherapy bone marrow aplasia is evident. In the case of accompanying myelofibrosis, fragmentation of reticular and collagen fibers is noticeable only following stem cell transplantation. The first precursors, which appear after the second or third week after transplantation are erythroid precursors found in small groups. Promyelocytes and myelocytes are observed between the second and fourth week, the ration off erythroid to myeloid populations at that time is 1:1. Megakaryocytes ate the last precursors to appear. Trephine biopsy evaluation is useful in diagnosis of graft rejection, especially in case of a discrepancy between the blood morphology examination and bone marrow cellularity. Granuloma formation is an unspecific finding, which requires diagnosis of infections. Residual disease can be recognized in histopathologic examination, or sooner, with immunophenotyping, cytogenetic examination using FISH or PCR. Secondary neoplasms to high dose radiotherapy or chemotherapy are usually lymphoproliferative processes connected with latent EBV infections, myelodysplastic syndromes and acute myeloblastic leukaemia. PMID- 10822997 TI - Gastrointestinal complications in bone marrow transplant patients. AB - Complications of bone marrow transplantation are numerous, multifactorial and may affect any organ in the body. The involvement of the gastrointestinal system represents one of dominant sites of complications. Clinical management of gastrointestinal complications can be challenging in these patients. This overview summarizes common bone marrow transplantation related conditions affecting esophagus, stomach, small and large intestine as well as liver. These are discussed primarily from the clinical point of view with emphasis on the differential diagnosis. The histomorphologic features of these conditions are discussed as well. PMID- 10822998 TI - [Adenocarcinoma of the pancreas. Palliative surgery]. AB - In case of unresectable adenocarcinoma of the pancreas, laparoscopy can allow to perform gastric and biliary by-pass which will improve the quality of life at a lower cost than conventional surgery for the patients. PMID- 10822999 TI - [Curative surgical treatment of pancreatic carcinoma]. AB - Surgery remains the unique curative treatment of the pancreatic carcinoma. The operative risk is actually low. The indication and limitations of surgical resection are discussed. PMID- 10823000 TI - [Pancreatic adenocarcinoma: role of pathologic anatomy]. AB - Duct cell adenocarcinoma is by far the most frequent neoplasm of the pancreas, accounting for about 80% of malignant tumors. In this report, we describe the main diagnostic pitfalls of this cancer in routine or frozen sections, as well as the help of immunohistology to make the differential diagnosis. We also discuss the potential interest of tumor markers for evaluating the prognosis of these tumors and provide information about the creation of a Web site (URL: http://www.ebm.lib.ulg.ac.be/index.htm) created to help clinicians exploit the laboratory results. PMID- 10823001 TI - [Role of radiotherapy in the multidisciplinary approach to pancreatic adenocarcinoma]. AB - Pancreatic tumors have a very bad prognosis even after complete surgery. Radiotherapy has an important role to play either postoperative with associated chemotherapy or palliative and alone. Presently 5-FU is the most widely used molecule but gemcitabine is getting more and more acceptance. Intraoperative radiotherapy but especially preoperative radiotherapy seem to be two promising techniques. PMID- 10823002 TI - [Has there been progress in the treatment of metastatic pancreatic cancer?]. AB - Pancreatic cancer is one of the most lethal cancers in the western world. The authors review the different chemotherapeutic regimens for the palliative approach of this disease. 5-fluorouracil has been the only drug with some efficacy for a longtime. Recently a clinical benefit has been obtained with gemcitabine. Polychemotherapy regimens based on gemcitabine, 5-fluorouracil, cisplatin and/or epirubicin have been administered in clinical phase II trials. The superiority of polychemotherapy in terms of palliative benefits or survival is uncertain. Palliative care remains an important issue. PMID- 10823003 TI - [Chronopharmacologic approach to the administration of anticancer agents in pancreatic adenocarcinoma. Pilot experience]. AB - The authors first analyzed the potential interest of the delivery of anticancer agents according to chronobiological concepts for human pancreatic cancer. They report their experience on 41 patients treated in adjuvant (12 cases) or palliative (29 cases) situations. The excellent therapeutic index observed warrants further evaluations of this concept in randomized trials. PMID- 10823004 TI - [Image of the month. Knotted umbilical cord]. PMID- 10823005 TI - [Revised postmenopausal treatments? A new polemic!]. AB - The study by Schairer et al. aims to determine whether increases in risk of breast cancer associated with the estrogen-progestin regimen are greater than those associated with estrogen alone. This study is a cohort of follow-up data for 1980-1995 from the Breast Cancer Detection Demonstration Project, a nationwide breast cancer screening program that involved 29 screening centers throughout the United States. A total of 46,355 postmenopausal women were followed. During follow up, 2,082 cases of breast cancer were identified. Increases in risk with estrogen only and estrogen-progestin only were restricted to use within the previous 4 years. The relative risk increased by 0.01 with each year of estrogen-only use and by 0.08 with each year of estrogen-progestin-only use among recent users. Among women with a Body Mass Index of 24.4 kg/m2 or less, increases in relative risk with each year of estrogen-only use and estrogen progestin-only use among recent users were 0.03 and 0.12, respectively. The authors conclude that the estrogen-progestin regimen increases breast cancer risk beyond that associated with estrogen alone. This study was largely commented in the lay media. Unfortunately the Belgian media introduced the confusion between the relative risk and the risk attributable to estrogen and estrogen-progestin. The aim of this manuscript is to precisely inform our colleagues, to analyze the Schairer study and to present the actual figures of risk associated with the use of estrogen and estrogen-progestin replacement therapy. Finally, we formulate some suggestions for the physician to whom the patient declares: "Did you read the negative effects of hormones?". What should we advice? PMID- 10823006 TI - [Epidemiology, physiopathology and treatment of a frequent ailment: tinea pedis]. AB - Tinea pedis is an infection of the stratum corneum caused by dermatophytes. Nowadays, fitness centers, saunas, subtropical swimming pools and sport-shoes are more and more fashionable and are certainly responsible for the increase of cases of tinea pedis as dermatophytes grow preferentially in warm and humid environments. The clinical aspect is affected by several factors such as the host reaction to the infection, the virulence of the infective strain, species, and some local and environmental factors. If a dermatophytosis is suspected, it is mandatory to validate the provisional diagnosis by demonstration of the fungus (by culture or/and microscopic examination) to avoid useless and expensive treatments. The new antifungal molecules are very effective, well tolerated and allow short course of therapy improving the compliance of the patient in clinical practice. Relapse of tinea pedis is very common but could be avoided by several simple preventive measures of hygiene. PMID- 10823007 TI - [The dosage of anti-GAD and anti-IA2 autoantibodies: an aid to the early diagnosis of type 1 diabetes]. AB - Diabetes mellitus is a frequent metabolic disease characterised by a complex and inconstant phenotypic expression that complicates the classification of patients and sometimes delays their optimal management. In that slowly progressive disease leading to severe and irreversible complications, the use of early and specific genetic, immunological and/or metabolic markers may help in the classification of diabetic patients and in the orientation of therapeutic strategies; furthermore, it is also an essential aid in the early screening of subjects at risk of developing the disease. The assessment of classical immunological markers, such as islet cell antibodies (ICA) or anti-insulin antibodies (IAA) has been recently completed by the screening of new promising markers such as GAD- and IA2 antibodies. The presence of these markers confirms the autoimmune component of the disease and thus supports the diagnosis of type 1 diabetes, even if clinical symptoms are absent or inconsistent. In addition, it represents a strong argument in favour of the initiation of specific immunological therapies to preserve B cell number and function. PMID- 10823008 TI - [Malacoplakia associated with a myelodysplastic syndrome]. AB - Malakoplakia is a rare chronic disease with pleomorphic presentations. It involves many organs, particularly the urinary and gastro-intestinal tracts. It is less common on the skin. Lesions result from impaired phagocytosis of bacteria by macrophages. Malakoplakia is sometimes related to immunodeficiencies. The diagnosis is usually made by histological examination. We report a case associated with a myelodysplastic syndrome. PMID- 10823009 TI - [How I investigate ... a broncho-pulmonary cancer by metabolic imaging (PET-18 FDG)]. AB - Continuing advances in PET imaging have resulted in an improved ability to evaluate thoracic malignancies. Published reports demonstrate that PET provides accurate noninvasive detection of malignancy that is useful in the characterization of a pulmonary solitary nodule and in the mediastinal or extrathoracic staging of known lung cancer. Preliminary studies suggest that PET may also be able to assess the therapeutic response after surgery or radiation therapy. PMID- 10823010 TI - [Pharma-clinics. Medication of the month. Glimepiride (Amarylle)]. AB - Glimepiride, commercialized in Belgium under the trade name of Amarylle by Aventis, is a new sulphonylurea compound which is indicated in the treatment of type 2 diabetes, after diet and exercise failure. It is available as 2 mg tablets. The initial doses is 1 mg, to be progressively increased up to 4 mg per day, if necessary, with a maximal daily dose of 6 mg. It is recommended to take glimepiride once a day, with the first main meal. Because of a particular binding of this sulphonylurea to the B cells of Langerhans pancreatic islets and, perhaps, of the presence of some extrapancreatic effects, both hypoglycaemic risk and circulating plasma insulin levels are lower with glimepiride than with glibenclamide, the reference sulphonylurea agent used in comparative clinical trials. PMID- 10823011 TI - [Clinical study of the month. After DCCT, the EDIC study]. AB - The "Diabetes Control and Complications Trial" (DCCT) demonstrated that intensive insulin therapy, by reducing HbA1c levels by about 2%, delays the onset and slows the progression of microangiopathic complications (by at least 50%) in patients with type 1 diabetes. The "Epidemiology of Diabetes Interventions and Complications" (EDIC) study recently showed that the reduction in the risk of progressive retinopathy and nephropathy resulting from intensive therapy during the DCCT not only persists, but is amplified for at least 4 years (reduction by about 80% when compared to diabetic patients previously treated with conventional therapy during the DCCT). PMID- 10823012 TI - [Pharmacoeconomic evaluation of pravastatin in coronary secondary prevention in patients with myocardial infarct or unstable angina pectoris. An analysis based on the LIPID Study]. AB - BACKGROUND: Secondary coronary prevention with lipid lowering drugs have become a major issue in health policy formulation due to the large upfront investment in drug therapy. The recently completed LIPID trial with pravastatin in secondary prevention immediately raise the question whether pravastatin might be cost effective in Switzerland. METHODS: We conducted a cost-effectiveness analysis from the perspective of third party payers. The following costs were included in the analysis: daily treatment costs of pravastatin, non fatal myocardial infarction, coronary bypass operations and stroke. Life years gained was obtained by applying the declining exponential approximation of life expectancy. All calculations were standardized to 1000 treated patients. RESULTS: The net costs of treating 1000 patients (i.e. drug costs minus the costs of sequelae and interventions) are Fr. 3.6 Mio. In addition, a total of 430 life-years may be saved through treatment. The corresponding cost-effectiveness of pravastatin treatment is Fr. 8341 (nominal) Fr. 6985 (discounted). CONCLUSIONS: The results suggest that the cost-effectiveness of pravastatin in secondary prevention lie well within the threshold of other commonly accepted medical interventions and may be considered an economically viable approach for secondary coronary prevention. PMID- 10823013 TI - [Improving communication between ambulatory and inpatient care. A pilot project]. AB - Medical care of our population has made dramatic progress during recent decades. However, problems arose from an increasing specialization and subspecialization on the one hand and from the complexity of care structures on the other hand. In light of scarce financial resources and of increasing demands regarding quality by both recipients and payers of medical care the analysis of diagnostic and treatment processes gained considerable attention. In this context, a lack of cooperation between ambulatory and in-patient care has been complained about for a long time. The aim of our pilot project introduced here was therefore to assess the cooperation between physicians in private practice and our psychiatric hospital by means of a random sample query. The results are communicated here. PMID- 10823014 TI - [Silent myocardial ischemia in hypertensive patients]. AB - Silent myocardial ischemia occurs in hypertensive individuals with a prevalence of approximately 35%. ST-alterations are triggered by a) hypertensive peaks and b) heart rate increase. Like in patients with coronary heart disease most ischemic events occur without angina. They are clinically silent. In daily practice silent myocardial ischemia may be detected by ECG under physical load or with 24 h Holter ECG-monitoring. The latter can detect ischemic events missed by ECG-monitored exercise tolerance. In hypertensive patients the simultaneous, ST triggered recording of ECG and blood pressure data is more meaningful. Patients with silent ischemia are at higher risk than individuals without. Angor is not as strong a determinant of risk as silent ischemia. Hypertensive patients without coronary artery disease (CAD) who have silent ischemia may even have a worse prognosis than those with known CAD. It is therefore important to substantiate the objective extent of silent ischemia by ST-analysis. If detected it has to be included into therapeutic considerations with the goal to prevent such episodes by antihypertensive treatment. PMID- 10823015 TI - ["I can't breathe"]. PMID- 10823016 TI - [Vasculitis and mononeuritis multiplex]. AB - In 1991, a 67 year old patient, suffering from an adenocarcinoma of the colon, had to undergo hemicolectomy. In the five following years, isolated metastases developed in the liver and have been surgically removed. In 1997, acute mononeuritis multiplex appeared and nerve biopsy revealed vasculitis. There were no signs of inflammatory rheumatic illness, collagenosis, cryglobulinemia, infection or hypersensitivity to medication. We have therefore postulated a relation between the vasculitis and the malignancy. Tumorous relapse could however not be detected at that time. Under immunosuppressive therapy, the neurological deficiencies receded partially, though the carcinoma of the colon reappeared with systemic metastases in the wake of the vasculitis. Diagnostic measures and the relationship between malignancy and vasculitis are discussed. PMID- 10823017 TI - [Morbid obesity without concomitant pathology]. PMID- 10823018 TI - [Vasopeptidase inhibition: a new mechanism of action--a new antihypertensive drug]. AB - The novel principle of vaso-peptidase inhibition is based on the simultaneous inhibition of two enzymes, the angiotensin converting enzyme (ACE) and the neutral endopeptidase (NEP) by a single drug. NEP participates in metabolism of various natriuretic and vasodilatating peptides produced by atria and ventricles of the heart and endothelium (arterial natriuretic peptide, brain natriuretic peptide and C-Type natriuretic peptide). The orally effective vaso-peptidase inhibitor omapatrilate has recently become available. This substance reduced blood pressure in hypertensive patients in a dose-dependent fashion. It is tolerated as well as ACE inhibitors and seems to ameliorate mainly systolic pressure. PMID- 10823019 TI - [Effects of written intervention on the number of laboratory studies per patient]. AB - In the context of economic measures in health care we followed over a period of nine months the consequences of a written intervention on the attitude of house staff to prescribe laboratory tests. Since it is well known that these tests have a major impact on health costs several studies have been conducted to test whether costs can be reduced without sacrifice of treatment quality. The study was undertaken in 1997. It had three phases of three months duration each: one for observation, one with the intervention and a follow-up phase. The laboratory tests requested by eleven physicians for their patients during the first month after the initial visit were analyzed. During the intervention--phase six physicians chosen at random were informed about their own average as well as that of the entire group (mean of the entire observation period of all physicians). Unexpectedly the hypothesis that the number of laboratory tests requested per patient would drop only in the group of informed physicians and should stay the same for the physicians without this intervention did not materialize. The number of performed tests dropped in both groups. PMID- 10823020 TI - [Therapy of acute lung failure--is there really something new?]. PMID- 10823021 TI - [An "exotic" lymphadenopathy: Kikuchi-Fujimotor histiocytic necrotizing lymphadenitis. Case report and discussion]. AB - The case of a 25 year old women with isolated inguinal lymphadenopathy and persistent fever caused by necrotizing inguinal lymphadenitis Kikuchi-Fujimoto is reported. Diagnostic and differential diagnostic aspects of this self-limited disorder with unclear pathogenesis are discussed. PMID- 10823022 TI - [Phenytoin-induced hypersensitivity reaction with liver failure]. PMID- 10823023 TI - Contact angle measurement on dental implant biomaterials. AB - Wettability may be one of the surface factors to be considered when selecting dental implant biomaterials. Contact angles of dental implant surface preparations influence wettability and tissue adhesion. The contact angles of eight implant surface preparations were determined in this study. Contact angles were measured by a tele-microscope equipped with a protractor eyepiece. Groups 1 to 6 had a Ti6Al4V substrate. Group 1 was metallurgically polished, group 2 was blasted with 180 microns Al2O3, group 3 was blasted with 710 microns Al2O3, group 4 was hydroxyapatite (HA) blasted (125 microns), group 5 had a Calcitite HA plasma-sprayed coating, and group 6 was coated with plasma-sprayed MP-1 HA. Group 7 was metallurgically polished commercially pure (CP) titanium (grade 1), and group 8 was etched CP titanium (grade 1). Contact angles were measured 30 times for each group with distilled water and glycerol, and the determinations were statistically analyzed. Mean contact angles for groups 1 to 8 were 65.5, 65.3, 62.5, 67.9, 46.6, 81.7, 58.5, and 69.0, respectively, when tested with distilled water, and 70.7, 68.3, 81.6, 75.4, 67.1, 70.7, 62.3, and 82.5, respectively, when tested with glycerol. Analysis of variance and Tukey's Honestly Significant Difference test (p = 0.05) demonstrated significant differences between group 5 and all other groups when groups were tested with distilled water and demonstrated no significant differences between groups 5 and 7 when groups were tested with glycerol. Surface preparation of implant biomaterials affects wettability. In this study, Ti6Al4V coated with Calcitite HA had the lowest contact angles and the best wettability. PMID- 10823024 TI - The role of the dental technician in fabricating the single-stage implant prosthesis. AB - With the initiation of the concept of osseointegration, the use of dental implants has become a predictable and frequently used addition to comprehensive planning and treatment of edentulous patients. Implantology poses challenges for the dental laboratory not present in any other existing specialty. Single-stage implant surgery represents such an innovation. PMID- 10823025 TI - The use of titanium plates for bone regeneration with root form implants: a case report. AB - In reconstruction of the partially and totally edentulous ridges that have bony defects due to old trauma or longstanding atrophy, it is necessary to reconstruct both the width and height of the alveolar ridge. This clinical case report covers bone regeneration prior to implant placement to achieve an aesthetic and functional base for prosthetic restoration. The focus of this report will be on bone regeneration, which does not depend on the utilization of a barrier membrane. PMID- 10823026 TI - A modified socket seal surgery with composite graft approach. AB - The contour of the residual ridge is reduced within 1 year by approximately 25% in width after the extraction of a natural tooth. The augmentation of a tooth socket after an extraction decreases the loss of available bone width for an endosteal implant. Grafting at the same time as the extraction has benefits from both a patient and doctor perspective. However, primary closure is more difficult, and may require the facial keratinized gingiva to be undermined and approximated on the crest of the ridge, or the use of membranes, which are exposed during the soft tissue healing. The modified socket seal surgery uses a technique described by Landsberg and couples his procedure with autologous bone harvested from the maxillary tuberosity. As a result, the tooth extraction socket may be augmented with autologous bone and connective tissue with a simplified approach at the same time as the extraction of a tooth. PMID- 10823027 TI - Case I--full arch case: maxilla endosteal root forms 16, 15, 14, 13, 24, 25, 26, and 27 and mandible endosteal root forms 36, 35, 34, and 46. PMID- 10823028 TI - Case II--maxilla bilateral sinus elevation: endosteal root form implants 15, 16, 24, and 26 and mandible endosteal root form implants 37, 36, and 46. PMID- 10823029 TI - Case III--full arch case: maxilla endosteal root forms 16, 15, 14, 13, 11, 21, 23, 24, 25, and 26 and mandible endosteal root forms 36, 34, 33, 41, 43, and 45. PMID- 10823030 TI - Central nervous system plasticity and persistent pain. AB - Nerve signals arising from sites of tissue or nerve injury lead to long-term changes in the central nervous system and contribute to hyperalgesia and the amplification and persistence of pain. These nociceptor activity-induced changes are referred to as central sensitization. Central sensitization involves an increase in the excitability of medullary and spinal dorsal horn neurons brought about by a cascade of events, including neuronal depolarization, removal of the voltage-dependent magnesium block of the N-methyl-D-aspartate (NMDA) receptor, calcium entry into neurons, phosphorylation of the NMDA receptor, a change in the cell's excitability, and an increase in synaptic strength. These changes also include activation of other ionotropic and metabotropic excitatory amino acid receptors, neuropeptides such as substance P, neurotrophins, and kinases involved in the phosphorylation process. Central sensitization occurs in trigeminal nociceptive pathways, and more robust neuronal hyperexcitability occurs following deep tissue stimulation than following cutaneous stimulation. By means of Fos protein immunocytochemistry, researchers have found that 2 distinct regions are activated: the subnucleus interpolaris/caudalis transition zone (Vi/Vc) and the caudal subnucleus caudalis. The latter exhibits changes very similar to those in the spinal dorsal horn, but the Vi/Vc zone likely is involved in autonomic nervous system processing and activation of the pituitary-adrenal axis. Descending systems are also an important component of the central sensitization process and provide the neural networks by which cognitive, attentional, and motivational aspects of the pain experience modulate pain transmission. These findings of nociceptor activity-induced neuronal plasticity have important clinical implications in the development of new approaches to the management of persistent pain. PMID- 10823031 TI - A unified concept of idiopathic orofacial pain: clinical features. AB - The main features of atypical facial pain, stomatodynia, atypical odontalgia, and masticatory muscle and temporomandibular joint (TMJ) disorders are compared in this article, which included a search of articles indexed in MEDLINE. The fact that their terminology has been the subject of many debates can be considered a consequence of taxonomic difficulties and uncertainties. Epidemiologic studies indicate marked female predominance for all types of idiopathic orofacial pain. There is also a difference in the age of maximal prevalence between masticatory muscle and TMJ disorders and the other entities. The clinical presentations display several symptoms in common. Pain is oral, perioral, or facial and does not follow a nervous pathway. It has been present for the last 4 to 6 months or has returned periodically in the same form over a period of several months or years. The pain is continuous, has no major paroxysmal character, and is present throughout all or part of the day. It is generally absent during sleep. Clinical, radiographic, or laboratory examination does not reveal any obvious organic cause of pain. There is also a frequent presence of certain psychologic factors, personality traits, or life events. Based on these shared characteristics, a unified concept is proposed. Each of these entities belongs to a group of idiopathic orofacial pain and could be expressed in either the jaws, the buccal mucosa, the teeth, the masticatory muscles, or the TMJ. PMID- 10823032 TI - Odontalgia in vascular orofacial pain. AB - A case of episodic, spontaneous odontalgia, aggravated by ingestion of cold food, with no apparent dental pathology is presented. Attempts at alleviating the pain by means of root canal treatment had failed in previous, similar episodes, and pain and pulpal hyperalgesia had shifted to other locations. Primary vascular orofacial pain was diagnosed and effective control obtained by prophylactic treatment with propranolol, a beta-adrenergic blocker. A prophylactic attempt with nifedipine, a calcium channel blocker, failed to alleviate the pain. This diagnostic entity and possible therapeutic approaches are discussed. PMID- 10823033 TI - Toothache of cardiac origin. AB - Pain referred to the orofacial structures can sometimes be a diagnostic challenge for the clinician. In some instances, a patient may complain of tooth pain that is completely unrelated to any dental source. This poses a diagnostic and therapeutic problem for the dentist. Cardiac pain most commonly radiates to the left arm, shoulder, neck, and face. In rare instances, angina pectoris may present as dental pain. When this occurs, an improper diagnosis frequently leads to unnecessary dental treatment or, more significantly, a delay of proper treatment. This delay may result in the patient experiencing an acute myocardial infarction. It is the dentist's responsibility to establish a proper diagnosis so that the treatment will be directed toward the source of pain and not to the site of pain. This article reviews the literature concerning referred pain of cardiac origin and presents a case report of toothache of cardiac origin. PMID- 10823034 TI - The anecdotal tradition and the need for evidence-based care for temporomandibular disorders. PMID- 10823035 TI - Epidemiology and treatment need for temporomandibular disorders. PMID- 10823036 TI - The neural basis of temporomandibular joint and masticatory muscle pain. PMID- 10823037 TI - The differential diagnosis of temporomandibular disorders. PMID- 10823038 TI - The prescription of diagnostic images for temporomandibular joint disorders. PMID- 10823039 TI - On the management of temporomandibular disorders: a plea for a low-tech, high prudence therapeutic approach. PMID- 10823040 TI - Developmental anomalies of the temporomandibular joint. PMID- 10823041 TI - Muscle-related temporomandibular disorders. PMID- 10823042 TI - Temporomandibular disorders: disc displacements. PMID- 10823043 TI - Posttraumatic temporomandibular disorders. PMID- 10823044 TI - Temporomandibular disorders: osteoarthritis. PMID- 10823045 TI - Surgical considerations in the management of temporomandibular joint and masticatory muscle disorders. PMID- 10823046 TI - Periodontal considerations in endo/perio lesions. PMID- 10823047 TI - The attitudes of New Zealand dentists and dental hygienists towards toothbrushes and toothbrushing. AB - The purpose of this study was to survey the attitudes of dentists and dental hygienists who practise in New Zealand, with respect to their personal habits, and the recommendations they give to their patients about toothbrush use. A questionnaire was mailed to 800 dentists and 74 dental hygienists with a response rate of 58% and 66% respectively. The results indicate that 76.6% of dentists and 89.1% of dental hygienists recommend toothbrush replacement every two to three months to their patients and generally follow this recommendation themselves. Both groups feel patients should replace their toothbrushes more often than they currently do and the majority tell their patients when to replace brushes. Dentists and dental hygienists have identified bristles that "no longer remove plaque" and "bent, splayed bristles" as the two most important indicators of when a new toothbrush is needed. Soft, compact head brushes are most often recommended by both groups. Most offices give toothbrushing instruction and distribute complimentary brushes. Dentists most often assume this task, but when a hygienist is employed, this duty is typically their responsibility. Oral B and Colgate toothbrushes are the brands most often recommended by both dentist and dental hygienists. PMID- 10823048 TI - Spontaneous alveolar bone sequestration. Case reports. PMID- 10823049 TI - The efficacy of a pre-procedural antiseptic mouthwash against bacterial aerosols. PMID- 10823050 TI - Melkersson-Rosenthal syndrome or oro-facial granulomatosis (OFG): an update. PMID- 10823051 TI - Case presentation: sialolithiasis. PMID- 10823052 TI - Implant update: fabrication of a surgical template. PMID- 10823053 TI - The effect of nicotine on growth of Streptococcus mutans. AB - There is little information available concerning the effects of nicotine on oral bacteria; and in particular, Streptococcus mutans, an important microbe in the etiology of dental caries. To test the effects of nicotine, Streptococcus mutans were incubated in either 0 or 10(-1)-10(-7) M concentrations of nicotine and then plated onto mitis-salivarius-bacitracin agar. Colonies were counted and treatment groups compared. Both 10(-1) and 10(-2) M nicotine caused total inhibition of bacterial growth, while 10(-3) and 10(-4) M produced significantly more colonies than control. Also, 10(-6) and 10(-7) M nicotine produced a significant reduction in the mean number of colonies. These results suggest a biphasic, dosage dependent effect of nicotine on the growth of Streptococcus mutans. Since 10(-3) M nicotine has been reported within the saliva of smokeless tobacco users, use of these products could stimulate growth of Streptococcus mutans and possibly place the user at increased risk for dental caries. PMID- 10823054 TI - Fabrication of a mandibular implant overdenture. PMID- 10823055 TI - Neurofibromatosis. PMID- 10823056 TI - Retained silicone impression material associated with a periodontal abscess. PMID- 10823057 TI - Root coverage using a subepithelial connective tissue graft and rotational flaps in the mandibular posterior region. PMID- 10823058 TI - Categories and treatment of unerupted teeth. PMID- 10823059 TI - Categories and treatment of unerupted teeth. PMID- 10823060 TI - The use of Alloderm and allograft to augment a mandibular arch for implant development. PMID- 10823061 TI - EMDOGAIN. PMID- 10823062 TI - Dental education in Canada: beyond 2000. PMID- 10823063 TI - Perception vs. reality. Two views of management service organizations. PMID- 10823064 TI - Sealing and dentin bond strengths of adhesive systems. AB - The objectives of this research were (1) to analyze the variations of the permeability of dentin after restoration with two polyacid-modified resin composites (Compoglass, Dyract) and four single-bottle adhesives (Prime & Bond 2.0, Syntac Single Component, OptiBond Solo, and Single Bond--Scotch Bond 1 in Europe--immediately (approximately 1 hour) after insertion. A perfusion system with distilled water was used at a pressure of 32.5 cm of water; (2) to study the bond strength of their interfaces; and (3) to find the correlation, if any, between both parameters. None of the materials used produced a complete cessation in fluid filtration. Tensile bond strengths were very low (maximum: P&B = 3.96 MPa) probably because of the very large bonding surfaces used (mean bonded surface area = 88.8 mm2). No significant correlation was found between tensile bond strength and the sealing ability for any material. PMID- 10823065 TI - Effect of bonded amalgam restorations on microleakage. AB - The purpose of this study was to evaluate quantitatively the effectiveness of adhesive dentin bonding systems in decreasing microleakage at the tooth-amalgam restoration interface. The results indicated that microleakage was significantly reduced when Amalgambond Plus or All-Bond 2 was used as liners in comparison to either Copalite varnish or no linear under amalgam restorations. No significant difference was found between the two dentin bonding systems at all time periods studied. PMID- 10823066 TI - Retention and shear bond strength of two post systems. AB - The purpose of this study was to compare the retention and shear strength of teeth restored with the Para Plus post (P) and the C post (C1 and C2) systems. Twenty-four P, C1, and C2 posts each were placed 7 mm into recently extracted and endodontically treated maxillary anterior roots and luted with Ketac-Cem and Bis Core respectively. In addition, 13 samples of each post type had a Bis-Core composite core placed. The tensile retention strength of the post and the shear strength of the post and core restorations when the core was loaded buccolingually at 45 degrees to the roots' long axis were determined at a strain rate of 1.3 mm/min. The C2 posts required significantly more tensile force (P < 0.0001) to remove them than the P or C1 posts. The shear strength of the post/core restorations was not significantly different (P < 0.04). The C1 restorations had 12 root fractures, the C2 restorations had 11 root fractures, and the P samples had six root fractures. It was concluded that the lack of stiffness of the C posts adversely affected the success of these restorations. PMID- 10823067 TI - Quantitative microleakage evaluation around amalgam restorations with different treatments on cavity walls. AB - The purpose of this in vitro study was to evaluate the dye penetration around amalgam restorations in dentin cavities by a quantitative test. Standardized circular cavities were prepared on the facial surface of 75 extracted human single-rooted teeth, and restored with dental amalgam. Different bonding/sealing treatments were used on the cavity walls before the restorations were placed. The specimens were thermo-cycled between 5 +/- 2 degrees C and 55 +/- 2 degrees C for 500 cycles with 1-minute dwell times, followed by immersion in a 2% methylene blue solution for 12 hours at 37 degrees C. The quantitative microleakage was evaluated by spectrophotometry and was expressed by microgram dye per tooth structure. The results showed that the use of bonded amalgam restorations was more effective in reducing microleakage. PMID- 10823068 TI - Bond strength of composite to dentin treated by air abrasion. AB - This study compared the bond strength of composite to dentin produced by an air abrasive dentin pretreatment and acid etch only. Two hundred sixty extracted human molars were randomly divided into 13 groups (n = 20). An occlusal dentin surface with a defined smear layer was exposed. Dentin was conditioned either with 37% H3PO4 for 20 seconds, or an air-abrasion unit (KCP 1000, ADT) was used with 120 and 160 psi pressure and two different particle sizes, 50 and 27 microns. A combination of both treatments beginning with air abrasion was tested as well as dentin specimens without pretreatment. The dentin adhesive Syntac Single-Component was applied under dry and moist conditions. Composite cylinders with a bond diameter of 2.5 mm were bonded in two increments to the teeth. Specimens were stored for 24 hours at 37 degrees C in distilled water and then thermo-cycled for 1000 cycles (5/55 degrees C). Shear bond strength was tested using a Universal Testing Machine at 0.5 mm/min cross-head speed. Means and standard deviations in MPa were calculated. Analysis of variance and Bonferroni pairwise comparison tests were performed. Lowest bond strengths were obtained in specimens without conditioning the dentin. Air-abraded specimens showed a significantly higher bond strength than the before-mentioned groups. The comparison for acid-etched groups and air-abraded specimens revealed higher values for the abraded groups. Air abrasion with 160 psi pressure produced a significantly higher bond strength than 120 psi. There was no significant difference between the 27 and 50 microns particles. Air abrasion in combination with a self-priming bonding agent produced bond strength on dentin at least comparable to conditioning with 37% H3PO4. PMID- 10823069 TI - Marginal adaptation of heat-pressed glass-ceramic veneers to Class 3 composite restorations in vitro. AB - The aim of the present in vitro study was to compare the marginal adaptation and integrity of heat-pressed glass-ceramic veneers to adjacent class 3 composite restorations and to enamel using four dual-curing composite resin cements of different viscosity with their corresponding dentin bonding agents. Thirty-six caries-free human maxillary incisors were first restored with mesial and distal class 3 composite restorations and then prepared for facial ceramic veneers. The cavity margins of the veneers were located either in the class 3 composite restorations or in the residual enamel. Heat-pressed glass-ceramic veneers (IPS Empress) were inserted adhesively using one of the following four luting systems in nine teeth: SonoCem (SC) with EBS; Variolink Ultra (VU), Variolink High Viscosity (VHV), and Variolink Low-Viscosity (VLV) with Syntac. The veneer margins in the region of the composite restoration and in the region apical to the composite restoration (ceramic/composite resin cement interfaces, composite resin cement/composite restoration interface, and composite resin cement/enamel interface) were evaluated before and after thermo-cycling and mechanical loading (TCML) by quantitative margin analysis under a scanning electron microscope (SEM) using an image analysis system. Furthermore, microleakage was assessed in each tooth by dye penetration after TCML. For all luting systems, SEM analysis revealed excellent marginal adaptation of the ceramic veneers to the composite restorations as well as to enamel. The median percentages of marginal gap formation were 1.1% and less before TCML and 5.1% and less after TCML. The error rates method revealed no statistical influence of the interface or of the viscosity of the luting material. Maximal values of dye penetration showed a significantly higher microleakage at veneers cemented with VU (median: 86.4%) compared to SC (median: 13.3%). In conclusion, the present data demonstrated that existing clinically acceptable class 3 composite restorations have no negative influence on the marginal adaptation of ceramic veneers. This was valid independent of the viscosity of the dual-curing composite resin cement when SC, VHV, or VLV was used. PMID- 10823070 TI - A method for mounting natural teeth in a commercial dentoform. PMID- 10823071 TI - Buonocore Memorial Lecture. Thoughts on contemporary restorative materials. PMID- 10823072 TI - Total bonding vs selective bonding: marginal adaptation of Class 2 composite restorations. AB - For improving the marginal integrity of composite restorations, a total bonding method has been advocated besides the conventional selective bonding method. Total bonding avoids the placement of a base. The entire internal cavity surface is available for the adhesive bond. Selective bonding involves the placement of a base covering the pulpal floor as well as the pulpoaxial wall. In this study, five dentin/enamel bonding system/composite combinations were used to restore 60 class 2 cavities with their cervical margins below the cementoenamel junction (CEJ); six teeth per material were restored according to the total bonding and the selective bonding methods. Before and after simultaneous thermo-cycling and mechanical loading (TCML) marginal adaptation was evaluated on replicas in the SEM. Microleakage was determined by dye penetration on the original samples after TCML. The data were statistically evaluated with the Mann-Whitney U-test and the Wilcoxon test. The error rates method was applied. In SEM analysis the error rates method indicated a significant difference between the two restoration methods in general. In the pairwise comparisons, no significant differences between the selective bonding and total bonding methods were found for Syntac/Tetric, Gluma 2000/Pekafill, and Gluma/Pekafill. With Scotchbond Multi Purpose (SBMP) and All-Bond 2 (AB2), total bonding revealed significantly (P < or = 0.01) less gap formation before and after TCML than selective bonding. Accordingly, total bonding showed significantly less (P < or = 0.01) dye penetration with Scotch-bond Multi-Purpose and All-Bond 2 compared to selective bonding. In conclusion, the reduction of microleakage by application of the total bonding method depended upon the bonding system used. Total bonding could be an alternative procedure for the adhesive restoration of class 2 cavities when their gingival margins are apical to the cementoenamel junction, provided the proper system is used and pulp damage is prevented. PMID- 10823073 TI - Water storage effect on the marginal seal of resin-modified glass-ionomer restorations. AB - The effect of storage in water on the marginal adaptation of glass-ionomer restorations placed in extracted human premolars and Teflon molds was studied. When the cavity preparations were filled with a glass ionomer and polished immediately after setting, a marginal gap of approximately 15 microns was formed. In contrast, when polishing following storage in water the marginal gap was 0 to 2 microns for the resin-modified and the conventional glass ionomers respectively. The bond strength of the glass ionomer to enamel and to dentin and the flexural strength of the glass ionomer increased after storage in water. Therefore the marginal seal of glass-ionomer restorations may not only be improved due to hygroscopic expansion during storage in water but also may exhibit greater bond strength. PMID- 10823074 TI - Evaluation of antibacterial activity of three dentin primers using an in vitro tooth model. AB - This study compared the antibacterial activities of three dentin primers and investigated a newly designed experimental system using a bovine tooth model method for evaluating antibacterial activity by comparing this method with a conventional disk diffusion method. The antibacterial activities of SA primer in Clearfil Liner Bond, LB primer in Clearfil Liner Bond II, and ED primer in Panavia 21 were evaluated using the conventional disk diffusion method. The 50 microliters aliquot of each primer was applied to three sterilized paper disks, then placed onto Tryptic Soy agar plates already inoculated with Streptococcus mutans. After anaerobic incubation for 48 hours, the diffusion of antibacterial components was determined using the inhibition zone produced around the paper disk. The diameter of the inhibition zones was measured and the average calculated. Standardized cavities (diameter 5.0 mm, depth 3.0 mm) were prepared on the labial surfaces of bovine teeth and inoculated with S mutans (10(6) CFU/microliter) following sterilization by 60Co gamma rays (50 KGy). The teeth were divided into four groups: SA primer, LB primer, ED primer, and a control group. Except for the control teeth, the cavity preparations were treated with the respective dentin primers, and then firmly sealed with a temporary sealing material. The teeth were placed in bottles containing melted Tryptic Soy agar. Five ml of Tryptic Soy broth was then added to the surface of the hardened Tryptic Soy agar. After 1 week's incubation of the teeth in the bottles at 37 degrees C, the number of bacteria remaining in each cavity was counted, except for eight specimens, which were used for SEM observation. The ED primer showed the widest inhibition zone in the disk diffusion test, which was significantly different from the other primers. Using the bovine tooth model, all dentin primers showed antibacterial activity, with significant differences found among the four groups. The results indicated that ED primer had the strongest antibacterial effect among the three primers. PMID- 10823075 TI - Antibacterial activity of glass-ionomer restorative cements exposed to cavity producing microorganisms. AB - The antibacterial activity of the glass-ionomer restorative cements Ketac-Fil, Ketac-Silver, Fuji II LC, and Vitremer was studied in vitro, in conjunction with a total of 32 strains of five bacterial genera that may be associated with dental caries: Streptococcus spp, Lactobacillus spp, Actinomyces spp, Porphyromonas spp, and Clostridium spp. Agar plate diffusion was the method used for the bacterial cultures, which included a chlorhexidine control. All four glass-ionomer cements were found to inhibit bacterial growth, though with noteworthy differences in their spheres of action. Vitremer was the cement determined to have the greatest antibacterial effects, whereas Ketac-Silver presented the least inhibitory action. PMID- 10823076 TI - Color of restorative materials after staining and bleaching. AB - This study determined the effect of a 10% carbamide peroxide bleaching agent on the removal of stain from restorative materials. Color changes (delta E*) of three restorative materials [compomer (Dyract); composite (TPH Spectrum); hybrid ionomer (Fuji II LC)] when exposed to juice/tea, chlorhexidine (CH), and water (control) for 120 hours were studied. Stained specimens were treated for two 2 hour periods with a bleaching agent (Platinum Tooth Whitening System) with and without the active ingredient. Color was measured at baseline, after staining, and after treatment using the CIE L*a*b* color system relative to CIE standard illuminant A (incandescent light) as measured by a reflection spectrophotometer. Means and standard deviations (n = 5) were calculated and data were analyzed by four-way ANOVA. All variables and interactions were statistically significant. Color changes caused by CH and water were not perceptible (delta E* < 3.3). After two 2-hour treatments, the following occurred with specimens stained with cranberry juice/tea: paste with and without active ingredient perceptibly changed color of stained composite. The stained hybrid ionomer perceptibly changed color after treatment with paste containing active ingredient but did not change after exposure to paste without active ingredient. The stained compomer was not perceptibly different with either treatment. Platinum successfully removed stains from the composite and hybrid ionomer tested. PMID- 10823077 TI - Fluoride release and antibacterial properties of new-generation tooth-colored restoratives. AB - The aim of this study was to compare the amounts and pattern of fluoride release and antibacterial properties of new-generation restoratives over a 35-day period. Materials evaluated included fluoride-releasing composites (Tetric, Experimental X), compomers (Dyract, Compoglass), and a resin-modified glass-ionomer cement (Fuji II LC). A conventional glass ionomer (Fuji II Cap) was used as a control for fluoride-release testing. Five samples of each restorative material were evaluated for daily fluoride release over a 35-day period by means of ion chromatography. Ranking of materials from least to greatest total fluoride release over 35 days was as follows: Tetric < Experimental X < Dyract < Fuji II LC < Compoglass < Fuji II Cap. Fuji II Cap had significantly greater fluoride release than all other materials evaluated. Fuji II Cap, Fuji II LC, and Compoglass had similar patterns of fluoride release characterized by a high initial release that was many times that released later. The fluoride-releasing composites evaluated stopped releasing fluoride by day 14. Antibacterial testing was conducted using the agar diffusion inhibitory test. Five samples of each restorative were assessed at baseline and weekly intervals up to 35 days. The microorganisms used were Lactobacillus casei, Streptococcus mutans, and Streptococcus sobrinus. IRM, a zinc oxide/eugenol cement, was used as the baseline control. None of the restorative materials evaluated affected the growth of L casei, S sobrinus, or S mutans at all time periods including baseline, where fluoride was detected in the agar beneath the specimen disks. There was no correlation noted between fluoride-release potential and antibacterial properties. PMID- 10823078 TI - Polymerization color changes of esthetic restoratives. AB - The color changes of three different types of tooth-colored restoratives during polymerization were investigated using colorimetry. L*, a*, b* color parameters of five different shades of Z100 (a mini-filled composite resin), Fuji II LC (a resin-modified glass-ionomer cement), and Dyract (a polyacid-modified composite resin) were taken precure and postcure. The results showed that the restoratives evaluated all underwent color changes during polymerization. The polymerization changes in color parameters were shade and not material dependent. Changes in L* parameter or lightness during polymerization were significant for all material and shade combinations and had the greatest influence on the overall polymerization color change. As the color change was perceivable by the human eye for most shades of materials, the clinical practice of polymerizing some material on, or adjacent to, the undried tooth to confirm shades of esthetic restoratives before restorative procedures is prudent. PMID- 10823079 TI - Digital radiology and image analysis for approximal caries diagnosis. AB - The aim of this study was to determine if radiology combined with digital reception and image analysis, radiovisiography (RVG), is effective in the early diagnosis of simulated approximal carious lesions. An experimental lesion similar to the one produced by caries was made in 39 permanent molars and premolars. Radiographic images of these lesions were obtained under standardized conditions using RVG. The image obtained was magnified 700 times and the densities of the lesion, the enamel, the dentin, and the pulp were measured using an image analysis program (Visualdent). The results showed statistically significant differences between the density of the produced lesions and that of healthy enamel. This measurement was independent of the differing thickness of the approximal enamel surface in molars and premolars. These results indicated the potential usefulness of the system tested in the diagnosis of incipient approximal caries. PMID- 10823080 TI - Effect of mechanical properties of resin composites on the efficacy of the dentin bonding system. AB - This study determined the relationship between marginal adaptation to dentin cavity preparations, tensile bond strength of the restorations, and mechanical properties of the composites. Contraction gaps, tensile bond strengths, flexural strengths, and Young's modulus of eight commercial resin composites were determined. Eight resin composites (Clearfil AP-X, Estelite, Estio LC, Litefil II A, Prodigy, Progress, Silux Plus, and Z-100) were applied to dentin cavities or flat dentin surfaces mediated with an experimental dentin bonding system consisting of 0.5 M EDTA dentin conditioner, priming with 35% glyceryl mono methacrylate solution, and a commercial dentin bonding agent application (Clearfil Photo Bond). The contraction gap of the resin composite in a cylindrical dentin cavity was prevented completely for three of the resin composites tested (Clearfil AP-X, Estelite, and Silux Plus). The measured tensile bond strength correlated significantly not only with the tensile strength (r2 = 0.506; 0.01 < P < 0.05), but also with the flexural strength (r2 = 0.871; P < 0.001) and Young's modulus (r2 = 0.712; 0.001 < P < 0.01) of the composites, whereas the contraction gap did not correlate significantly with the measured tensile bond strength, the tensile strength, the flexural strength, or Young's modulus (P > 0.05). However, the results did indicate that the higher tensile bond strengths measured in the traditional test may be related to the higher mechanical properties of the resin composites because of the number of specimens that exhibited cohesive failure. PMID- 10823081 TI - Evaluation of different methods for cleaning and preparing occlusal fissures. AB - The effectiveness of different methods for cleaning and preparing occlusal fissures before placing sealants was evaluated. Extracted mandibular molars received such treatments as brushing, pumicing, bur preparing, and air abrasion before application of fissure sealants. FluroShield fissure sealant was then applied to the occlusal fissures. Specimens were subjected to thermo-cycling and then immersed in a 10% solution of methylene blue, and finally sectioned. The sections were examined and photographed in a stereomicroscope, and the dye penetration was recorded using a scoring system. The results indicated that only the control (brushing with a dry brush) and the pumicing groups demonstrated dye penetration to the base of the sealant. Teeth prepared with the #1/4 round bur and air abrasion demonstrated a better seal in evaluated fissures. For this study, those three groups (occlusal fissures prepared with the #1/4 round bur and two air abrasion methods), demonstrated significantly better sealing (P < 0.01) than the control group and the other groups tested. PMID- 10823082 TI - Cuspal deflection of maxillary premolars restored with bonded amalgam. AB - The aims of this study were to measure cuspal deflection of premolars restored with bonded amalgam and to investigate bond resistance to thermo-cycling and cyclic loading. Strain gauges were used to measure cuspal deflection of maxillary premolars restored with MOD bonded amalgam restorations. A nondestructive method was used in which teeth were loaded repeatedly to record cuspal deflection following different restorative procedures. Ten extracted premolars with similar dimensions were selected and their roots mounted in resin bases 2 mm below the CEJ. Two single-element strain gauges were bonded to the buccal and lingual surfaces of the cusps of each tooth at a level that corresponded to the pulpal floor of MOD cavities. These were connected to a strain indicator with a built-in wheat-stone bridge. An Instron machine was used to apply a 100 N compressive load. Micro-strain readings were recorded with each loading at the following stages: (1) sound unprepared teeth (baseline reading), (2) following preparation of a medium-size MOD cavity, (3) 24 hours following restoration with amalgam, (4) following amalgam removal, (5) 24 hours following restoration with bonded amalgam. Durability of the bond was further tested by cyclic loading of 2000, 4000, 6000, and 8000 load cycles. Mean micro-strain values recorded at the buccal cusp were: 48.0 (21.6), 126.8(57.2), 121.4(53.3), 120.8(56.1), and 65.2(36.5) for test stages 1, 2, 3, 4, and 5 respectively. Cuspal deflections following cyclic loading recorded at the buccal cusp were: 60.0(41.0), 63.6(51.9), 59.6(36.3), and 61.6(36.8) at the above four cyclic loading stages respectively. A similar trend was also observed for measurements of the lingual cusp. It was concluded that bonding amalgam restorations decreases cuspal deflection and consequently may assist in restoring tooth strength under conditions of the oral environment. PMID- 10823083 TI - Influence of two dentin bonding systems on the demineralization of the root surface. AB - It has been assumed that dentin adhesives can prevent root surface caries. The aim of this study was to determine the caries-protective effect of two different dentin bonding systems on the demineralization of root surfaces in vitro. The root surfaces of 60 freshly extracted caries-free human molars were thoroughly cleaned and polished, thereby removing the cementum. The teeth were then coated with acid-resistant nail varnish, exposing two rectangular windows of 6 mm2 each. One window served as an untreated control, while the other window was treated with a dentin bonding system. The specimens were distributed among the following experimental groups--Group 1: Syntac, Heliobond (no air thinning); Group 2: Syntac, Heliobond (as recommended); Group 3: Syntac, without Heliobond; Group 4: Prime & Bond 2.0 (no air drying); Group 5: Prime & Bond 2.0 (as recommended); Group 6: Prime & Bond 2.0 (dentin pretreated with 36% phosphoric acid). Subsequently, all specimens were demineralized for 6 days with acidified gel (HEC, pH 4.8, 37 degrees C). From each tooth, three dentinal slabs were cut perpendicular to the polished surface of the windows. The slabs were ground to a thickness of 80 microns and imbibed with water. The depth of the respective demineralized areas was determined using a polarized light microscope. All control specimens exhibited lesions with a mean depth of 67 microns. In Groups 2, 3, and 5 the lesion depth was reduced significantly, while in Groups 1, 4, and 6 no lesions could be detected. It was concluded that the demineralization of the root surface can be impeded by application of the dentin adhesives tested. PMID- 10823084 TI - Effect of saliva contamination on the bond of dentin to resin-modified glass ionomer cement. AB - This in vitro study compared the shear bond strength of a resin-modified glass ionomer restorative material (Fuji II LC) bonded to saliva-contaminated dentin versus non-contaminated dentin. Seventy-five extracted human molar teeth were randomly divided into five groups of 15 samples each. The dentin was treated with 10% polyacrylic acid for 20 seconds, rinsed, and dried. The acid-treated dentin surfaces in Groups 1-4 were contaminated with saliva. In Group 1, the saliva was air thinned. In Groups 2-4, saliva was dried completely with compressed air. The saliva-contaminated dentin in Group 3 was rinsed and dried. The saliva contaminated dentin in Group 4 was rinsed, dried, treated with 10% polyacrylic acid, and dried. Specimens in Group 5 received no contamination. The resin modified glass-ionomer cement restorative material was mixed and applied to the dentin surfaces. Following placement of the restorative material and 7 days of storage, the specimens were thermo-cycled 300 times. Using the Instron Universal Testing Machine, a shear force was applied to the restorative material. Shear bond strength values were compared among the groups using a one-way ANOVA and Student-Neuman-Keuls Multiple Range Test (alpha = 0.05). The non-contaminated specimens (Group 5) were significantly stronger than the contaminated specimens (Groups 1-4). There were no significant differences in bond strength among the groups containing contaminated specimens. Salivary contamination occurring after dentin etching significantly reduced the bond strength of the resin-modified glass-ionomer restorative material to dentin. Neither rinsing nor rinsing and re etching resulted in bond strengths as great as to non-contaminated dentin. PMID- 10823085 TI - Human-eye versus computerized color matching. AB - This project compared the difference in color matching between human-eye assessment and computerized colorimetry. Fifty dental personnel were asked to color match Vita Lumin shade tabs to seven different randomly arranged test tabs from the Z100 shade guide. All evaluators were blinded to the shades of the test tabs and were asked to match only body shade of the Vita Lumin tab to the middle third or body of each test tab. The results obtained were subsequently computed into L*a*b* values and compared with results obtained by computerized colorimetry. Results indicate that the difference in color matching between human eye assessment and computerized colorimetry is shade dependent. Discrepancy was significant for b* coordinates for shades A1 and B2 and L* and b* coordinates for shade C4. For all shades evaluated, color difference between human-eye and computerized color matching is perceivable under clinical settings, as delta E values are greater than 3. There is a need for correction factors in the formal specification of the color-matching software due to the discrepancy between human eye and computerized colorimetric color matching. PMID- 10823086 TI - Cuspal reinforcement in endodontically treated molars. AB - This in vitro study compared the ability of horizontal pins and a dental adhesive to reinforce the facial cusps of endodontically treated mandibular molars. Seventy-two mandibular molars were divided into six groups and mounted in acrylic blocks (n = 12). In Groups 1-5 standardized endodontic access and instrumentation in the coronal one-third of each root canal were completed. In Groups 1-4 the lingual cusps were reduced, leaving the buccal cusps intact. The facial cusps of the teeth in each group received one of the following modes of reinforcement: Group 1--no reinforcement; Group 2--dentin adhesive (Amalgambond Plus); Group 3- two horizontal TMS Minim pins; Group 4--two horizontal TMS Minim pins and Amalgambond Plus. Teeth in Group 5 were prepared for and restored with a complete cuspal coverage amalgam restoration using four vertical TMS Minim pins. Group 6 consisted of intact natural teeth. Using an Instron Universal Testing Machine, the lingual slope of the facial cusp of each specimen was loaded to failure using a compressive force applied at an angle 60 degrees to the long axis of the tooth. The mean fracture strengths for all groups were analyzed using a one-way ANOVA and Student-Newman-Keuls multiple range test (alpha = 0.05). Fracture patterns and modes of failure were also evaluated. RESULTS: The intact teeth (Group 6) were significantly more fracture resistant than all other groups, with the exception of Group 4 (combination of pins and adhesive). Group 1 (non-reinforced teeth) was significantly weaker than all other groups. Groups 2-4 (specimens with reinforced cusps) were not significantly different from each other. The use of horizontal pins or a combination of horizontal pins plus dentin adhesive for cuspal reinforcement resulted in significantly more teeth demonstrating favorable fracture patterns than did the use of adhesives alone. CONCLUSION: The buccal cusps of endodontically treated mandibular molars reinforced with a combination of horizontal pins and dentin adhesive were not significantly weaker than intact teeth. Of the restored teeth, those which had buccal cusps reinforced with horizontal pins and those treated with complete cuspal coverage amalgam restorations exhibited the most favorable restorative prognosis following cusp fracture. PMID- 10823087 TI - Effects of aging on repair bond strengths of a polyacid-modified composite resin. AB - The effect of age of a poly-acid-modified composite resin on repair bond strength after different methods of surface conditioning was studied. Surface conditioning methods included the following: maleic acid with resin application; polyacrylic acid with resin application; sand-blasting with resin application. Shear bond testing between the aged and new material was carried out with an Instron Universal Testing Machine. Although repair bonds strengths after all surface conditioning methods were significantly higher than the control group at 1 week, no statistically significant differences in bond strengths were noted after aging the material for 6 months. After all aging periods, surface conditioning with sand-blasting and resin application resulted in the highest repair bond for poly acid-modified composite resins. Specimens with cohesive failure in the material gave significantly higher repair bond strengths than specimens with adhesive failure at the repaired interface. PMID- 10823088 TI - Amalgam restorations: postoperative sensitivity as a function of liner treatment and cavity depth. AB - The purpose of this clinical study was to assess the sensitivity reported by patients following the insertion of class 1 or class 2 amalgam restorations in the treatment of primary carious lesions of different depths. Ninety subjects with previously untreated teeth requiring restorations due to caries lesions were selected: 32 teeth had lesions that were clinically and radiographically judged to be located in the outer one-third of dentin, 30 were located in the middle one third of dentin, and 28 were located in the inner one-third of dentin. Four different lining regimens were employed: Group 1--no linear; Group 2--two coats of Copalite liner; Group 3--a dentin adhesive resin liner (Scotchbond Multi Purpose); Group 4--a resin-modified glass ionomer (Fuji Bond LC). Patients were contacted on days 2 and 7 postoperatively and questioned regarding the presence or absence of sensitivity, the stimuli that created the sensitivity, if any, the duration of any sensitivity, and the intensity of any sensitivity using a rating from None to Severe. If sensitivity was experienced on day 7, patients were also contacted on days 14, 30, and 90 to assess the sensitivity at those intervals. The chi-square test of independence showed no significance at the 0.05 level between the different dentin treatments and cavity depths. By day 2, 19% of lesions located in the outer one-third, 27% of lesions located in the middle one third, and 29% of lesions located in the inner one-third of dentin were sensitive. On day 30, four teeth were still sensitive, two located in the middle one-third and two located in the outer one-third of dentin. On day 90 all teeth were without sensitivity. PMID- 10823089 TI - Hot air or full steam ahead? An empirical pharmacokinetic model of potent inhalational agents. PMID- 10823090 TI - Assessment of clinical competence. PMID- 10823091 TI - 2000--time to apply Occam's razor to failure of hypoxic pulmonary vasoconstriction during one lung ventilation. PMID- 10823092 TI - Pharmacokinetics of inhaled anaesthetics in a clinical setting: description of a novel method based on routine monitoring data. AB - Pharmacokinetic parameters of inhaled anaesthetics have previously been assessed experimentally in healthy volunteers. In contrast, we developed a method to estimate pharmacokinetic parameters under clinical conditions. We obtained data from the continuous routine monitoring of fractional concentration and ventilation during anaesthesia with desflurane, isoflurane and sevoflurane. By simulation studies, we assessed the effects of several sources of variation, including the noise of measurement, the second gas effect and rounding errors or a limited number of displayed digits. Stable fits to a two-compartment model were obtained for both real and simulated data sets in all cases. The most stable parameter was the intercompartmental clearance, and the most sensitive parameter was the volume of distribution. The bias in pharmacokinetic parameters caused by adding errors to measured concentrations was similar for the different compounds. We conclude that the model allows the estimation of an alternative set of pharmacokinetic parameters that can reliably describe the behaviour of volatile anaesthetics under clinical conditions, and allow comparison between agents. PMID- 10823094 TI - General versus regional anaesthesia for hip fracture surgery: a meta-analysis of randomized trials. AB - Hip fracture surgery is common and the population at risk is generally elderly. There is no consensus of opinion regarding the safest form of anaesthesia for these patients. We performed a meta-analysis of 15 randomized trials that compare morbidity and mortality associated with general or regional anaesthesia for hip fracture patients. There was a reduced 1-month mortality and incidence of deep vein thrombosis in the regional anaesthesia group. Operations performed under general anaesthesia had a reduction in operation time. No other outcome measures reached a statistically significant difference. There was a tendency towards a lower incidence of myocardial infarction, confusion and postoperative hypoxia in the regional anaesthetic group, and cerebrovascular accident and intra-operative hypotension in the general anaesthetic group. We conclude that there are marginal advantages for regional anaesthesia compared to general anaesthesia for hip fracture patients in terms of early mortality and risk of deep vein thrombosis. PMID- 10823093 TI - Pharmacokinetics of inhaled anaesthetics in a clinical setting: comparison of desflurane, isoflurane and sevoflurane. AB - The pharmacokinetic characteristics of desflurane, isoflurane and sevoflurane (16 patients for each anaesthetic) were estimated from measurements of inspired and end-expired agent concentrations and ventilation, obtained during routine anaesthesia in patients undergoing maxillofacial surgery (mean age 38 yr, duration of anaesthesia approximately 2 h). A two-compartment model described the data adequately. Although isoflurane and sevoflurane have almost the same tissue/blood partition coefficients, significant differences between substances were observed for the peripheral volume of distribution (medians and ranges: desflurane, 612 (343-1850) mlvapour kgbw-1; isoflurane, 4112 (1472-9396) mlvapour kgbw-1; sevoflurane, 1634 (762-8843) mlvapour kgbw-1) and the transport clearance from the central to the peripheral compartment (desflurane, 7.0 (4.4-11.1) mlvapour kgbw-1 min-1; isoflurane, 30.7 (15.9-38.7) mlvapour kgbw-1 min-1; sevoflurane, 13.0 (9.8-22.4) mlvapour kgbw-1 min-1). Thus, during clinical anaesthesia the important characteristics of the compounds could be obtained and compared between substances from simple data. PMID- 10823095 TI - Novel bioadhesive delivery system for percutaneous local anaesthesia. AB - We have assessed the efficacy of a novel bioadhesive amethocaine patch device, compared to Ametop gel, in a randomized, double-blinded trial. Patch and gel formulations, including placebos, were applied to the forearms of volunteers (n = 30) for 40 min. Once the formulations were removed from the skin, anaesthesia was assessed by volunteers using a conventional pinprick test. Pain scores were recorded for 4 h after removal of gels and patches. Statistical analysis of the results indicated that both amethocaine gel and patch preparations were superior to placebo (P < 0.05). No significant difference was observed between amethocaine gel and patch formulations (P > 0.05) in either onset time or duration of action for percutaneous local anaesthesia. The results of this study indicate therefore that the novel bioadhesive patch provides clinically comparable anaesthesia to the established gel formulation in a more defined dosage form. PMID- 10823096 TI - Prophylactic antiemetic effect of dexamethasone in women undergoing ambulatory laparoscopic surgery. AB - The aim of this study was to evaluate the prophylactic antiemetic effect of i.v. dexamethasone in women undergoing ambulatory laparoscopic tubal ligation. Ninety patients requiring general anaesthesia for laparoscopic tubal ligation were enrolled in a randomized, double-blind, placebo-controlled study. Forty-five patients received dexamethasone 10 mg i.v. and 45 received saline 2 ml i.v. at the induction of anaesthesia. We found that 27% of patients in the dexamethasone group, compared with 63% of patients in the saline group, reported nausea and vomiting within 4 h after surgery (P < 0.01). Seven per cent of patients in the dexamethasone group, compared with 28% of patients in the saline group, received a rescue antiemetic (P < 0.05). During the 24-h postoperative observation period, 34% of patients in the dexamethasone group, compared with 73% of patients in the saline group, reported nausea and vomiting (P < 0.001). We conclude that dexamethasone 10 mg significantly decreases the incidence of postoperative nausea and vomiting in women undergoing ambulatory laparoscopic tubal ligation. PMID- 10823097 TI - Acupressure and prevention of nausea and vomiting during and after spinal anaesthesia for caesarean section. AB - The efficacy of acupressure at the P6 point in the prevention of nausea and vomiting during and after Caesarean section was studied. A double-blind, randomized controlled study of acupressure vs placebo was designed. Ninety-four patients scheduled for Caesarean section were included. The anaesthetic technique and postoperative analgesia were standardized. The use of acupressure reduced the incidence of nausea or vomiting from 53% to 23% compared with placebo (95% confidence interval (CI) 0.34-0.25; P = 0.002) during the operation and from 66% to 36% compared with placebo (95% CI 0.34-0.19; P = 0.003) after the operation. Other variables were similar between the groups. PMID- 10823098 TI - Continuous measurements of microcirculatory blood flow in gastrointestinal organs during acute haemorrhage. AB - Hypoperfusion of splanchnic organs is an important contributor to the development of multiple organ failure after major surgery and trauma. During general anaesthesia and surgery we compared changes in systemic haemodynamics and regional blood flow with changes in the distribution of microcirculatory flow (MBF) in multiple splanchnic organs in pigs exposed to acute haemorrhage. Seven pigs (25 kg) were bled to a mean arterial pressure of 40 mm Hg; 180 min later the shed blood was retransfused. MBF was measured in the intestinal mucosa (stomach, jejunum, colon), pancreas, liver and kidney using a six-channel laser Doppler flowmeter. Cardiac output was measured by thermodilution and superior mesenteric artery flow by ultrasonic flowmetry. During haemorrhage, MBF in the gastric and colon mucosa and flow in the liver and kidney decreased to a similar extent to regional and systemic flows (30-50%). In contrast, MBF in the jejunal mucosa remained virtually unchanged and flow in the pancreas decreased significantly more than systemic and regional flows (60%, P < 0.05). We conclude that: (1) changes in the distribution of MBF in the gastrointestinal tract during acute haemorrhage are heterogeneous and cannot be predicted from changes in systemic or regional haemodynamics; (2) MBF in the jejunal mucosa did not decrease during haemorrhage, indicating that autoregulation of blood flow in the mucosa remained intact during shock; and (3) MBF in the pancreas decreased significantly more than systemic and regional flows during shock, suggesting that the pancreas is particularly vulnerable to haemorrhage. PMID- 10823099 TI - P2X receptors in sensory neurones. AB - P2X receptors are a family of ligand-gated ion channels responsive to ATP. Seven subtypes have been identified which form homo-multimeric or hetero-multimeric pores. P2X3 receptors are selectively expressed predominantly on small-diameter nociceptive sensory neurones in the dorsal root, trigeminal and nodose ganglia, particularly the non-peptidergic subpopulations labelled with the lectin IB4. P2X2/3 labelling is also present in inner lamina II of the spinal cord and in sensory nerve projections to skin and viscera, but few receptors are present in skeletal muscle. P2X3 receptors are down-regulated after peripheral nerve injury and their expression can be regulated by glial cell-derived neurotrophic factor. P2X receptor activation of sensory neurones has been demonstrated in in vivo pain models, including the rat hindpaw and knee-joint preparations, as well as in inflammatory models. P2X4 and/or P2X6 receptors in the CNS also seem to be involved in pain pathways. Non-nociceptive P2 receptors on sensory nerves are present in muscle and on sensory endings in the heart and lung that initiate reflex activity involving vagal afferent and efferent nerve fibres. The sources of ATP involved in nociception and non-nociceptive sensory nerve stimulation are discussed as well as a novel hypothesis about purinergic mechanosensory transduction. PMID- 10823100 TI - Randomized placebo-controlled trial to assess the effect of remifentanil and propofol on bispectral index and sedation. AB - The effect of the combination of opiates and hypnotics on bispectral index (BIS) is unclear. This double-blind placebo-controlled trial investigated the effect on BIS and sedation of different infusion doses of remifentanil combined with a steady infusion of propofol. Forty patients initially received a target controlled infusion of propofol 2 micrograms ml-1 for 15 min. They were then randomized to receive either placebo, 0.01, 0.05 or 0.1 microgram kg-1 min-1 remifentanil for a further 15 min. We found a significant correlation between the dose of remifentanil and the change of BIS after 15 min of infusion. The correlation between all the sedation scores and their corresponding BIS was also significant. We concluded that remifentanil, in combination with propofol, reduces BIS when used for sedation. PMID- 10823101 TI - Role of dominant hand position during external cardiac compression. AB - Nineteen previously trained resident anaesthetists were instructed to perform adult single-rescuer basic life support for 5 min on a manikin, in a double-blind crossover design, changing the hand of contact with the sternum from right to left while performing external cardiac compression (ECC). Total, correct and incorrect ECCs comprising of inadequate depth, wrong hand placement, incomplete relaxation and too much compression were recorded and grouped according to the dominant hand (group DH) or non-dominant hand (group NH) in contact with the sternum. The number of correct ECCs was significantly greater in group DH, median 141 compared to group NH, median 97; P < 0.005. More ECCs were of inadequate depth in group NH, median 34 as compared to a median of 8 in group DH; P < 0.005. Similarly, the incidence of wrong hand placement was significantly higher in group NH; median of 4 versus median of 0 in group DH, P < 0.05. The incidence of incomplete relaxation and too much ECC was not significantly different between the two groups (P < 0.05). We conclude that ECC is performed with fewer errors when the dominant hand of the rescuer is placed in contact with the sternum. PMID- 10823102 TI - Epidural top-up solutions for emergency caesarean section: a comparison of preparation times. AB - We compared the preparation times of three solutions commonly used for epidural top-up for emergency Caesarean section. Twenty-two anaesthetists were asked to prepare fresh solutions in random order as quickly as possible: 0.5% bupivacaine 20 ml (B); 2% lidocaine 20 ml with 1:200,000 epinephrine (LE); and 0.5% bupivacaine 10 ml and 2% lidocaine 10 ml with 1:200,000 epinephrine and 8.4% sodium bicarbonate 2 ml (BLEB). Preparation times for B were approximately half of those for LE, which in turn were approximately half of those for BLEB (P = 0.0001). If local anaesthetic solutions with additives such as epinephrine or bicarbonate are prepared just before emergency Caesarean section, any possible reduction in onset time that they might afford may be offset by the additional preparation time required. PMID- 10823103 TI - In vitro validation of gastric air tonometry using perfluorocarbon FC 43 and 0.9% sodium chloride. AB - Monitoring splanchnic perfusion by means of gastric intramucosal tonometry is carried out using saline, or more recently, air tonometry using the Tonocap. The objective of this study was the validation of the Tonocap in saline and perfluorocarbon FC 43. The two methods underestimated the predefined PCO2 value by an average of 10%, with clinically acceptable precision. Accuracy of the Tonocap improved at high PCO2 values (9.33 and 9.94 kPa), whereas saline tonometry was superior at low PCO2 values (3.99 and 3.75 kPa). The Tonocap provides a fast and reliable estimation of the PCO2, and with the revised software requiring only 10 min of equilibration, will increase the comparability of future studies. PMID- 10823104 TI - Malignant hyperthermia in infancy and identification of novel RYR1 mutation. AB - Malignant hyperthermia (MH) has been reported as non-existent in children less than 1 yr old, although several unconfirmed reports have been published. A case report of MH in a 6-month-old child is presented, with confirmation of MH susceptibility by in vitro contracture testing of quadriceps muscle at 13 yr old. Genetic analysis revealed a novel RYR1 mutation that substitutes arginine 2452 for tryptophan in a region of the calcium channel mutated in several other MH pedigrees. PMID- 10823105 TI - Intratracheal recombinant human deoxyribonuclease in acute life-threatening asthma refractory to conventional treatment. AB - Recombinant human deoxyribonuclease (rhDNase) is a mucolytic agent used to relieve peripheral airway obstruction in patients with cystic fibrosis. We report dramatic sustained improvement following the intratracheal administration of rhDNase to a 3-yr-old boy with acute life-threatening asthma in whom 48 h of aggressive therapy had failed. PMID- 10823106 TI - Monitoring equipment induced tachycardia in patients with minute ventilation rate responsive pacemakers. AB - Minute ventilation-sensing pacemakers enable the paced heart to respond to an increased workload. Two patients with such a pacemaker developed pacemaker-driven tachycardia when connected to an electrocardiogram (ECG) monitor also capable of documenting ventilatory frequency and ECG lead disconnection. This tachycardia stopped when the ECG leads were removed. These pacemakers and monitors emit a low amplitude electrical current and measure the resultant impedence signal across the chest. When patients are connected to the monitor the pacemaker sensor summates both impedence signals and the paced heart rate is increased as a result. PMID- 10823107 TI - Stunned myocardium after hip arthroplasty. AB - We report a case of ST-segment elevation occurring in all leads of the ECG during hip arthroplasty. It is thought that this resulted from a stunned myocardium because wall motion abnormalities were reversible, there was no evidence of fixed or vasospastic coronary occlusion and there was only a slight increase in serial cardiac enzymes. Treatment with nicorandil improved the patient's cardiac function. A [123I]MIBG test revealed a high myocardial washout rate, suggesting that the stunned myocardium was caused by exposure to excessive norepinephrine induced by anaesthesia or surgery. PMID- 10823108 TI - Spinal cord infarction and tetraplegia--rare complications of meningococcal meningitis. AB - A previously healthy 25-yr-old female developed flaccid areflexic tetraplegia, with intact cranial nerve function, 36 h after the diagnosis of bacterial meningitis. Polymerase chain reaction studies of cerebrospinal fluid and blood were positive for Neisseria meningitidis, serogroup B. Magnetic resonance of the cervicothoracic spine revealed increased signal intensity and expansion in the lower medulla, upper cervical cord and cerebellar tonsils. Neurosurgical consultation recommended hyperventilation, dexamethasone and regular mannitol therapy rather than decompressive intervention. The clinical course over the following 12 days was complicated by the development of progressive central nervous and multisystem organ failure with disseminated intravascular coagulopathy. Autopsy revealed cerebral oedema with cystic infarction extending from the medulla to the upper cervical cord and cerebellar tonsils. Flaccid areflexic tetraplegia with spinal cord infarction has not been reported following bacterial infection in an adult. The clinical implications would suggest complete central nervous system evaluation of patients recovering from meningococcal meningitis, since spinal cord lesions, although uncommon, do occur. In those very rare situations where a patient develops significant peripheral neurological deficits, urgent magnetic resonance imaging is warranted, to rule out an infective focus or an underlying anatomical anomaly. PMID- 10823109 TI - Cranial subdural haematoma associated with dural puncture in labour. AB - A 23-yr-old primagravida sustained a dural puncture during epidural catheter insertion and developed a headache that settled with oral diclofenac and codydramol. On the third day after delivery, she convulsed twice without warning. As plasma urate was increased, the putative diagnosis of an eclamptic fit was made, and magnesium therapy was started. A contrast CT scan revealed that the cause of the patient's symptoms was a subdural haematoma with raised intracranial pressure. A coincidental arteriovenous malformation was noted. This case emphasises the need to consider the differential diagnoses of post-partum headache. The management of acute intracranial haematoma is described. PMID- 10823110 TI - Delayed bilateral internal carotid artery thrombosis following accidental strangulation. AB - A 24-yr-old male presented after a fishing accident in which he was pulled underwater by a rope attached to a crayfish pot. He was winched out of the water with the rope still around his neck, sustaining serious neck injuries that ultimately led to his death. After initial resuscitation, he remained fully conscious for approximately 8 h, after which there was a rapid and sudden deterioration in his level of consciousness. The presentation, investigation, management and subsequent postmortem findings are presented and discussed. PMID- 10823111 TI - Watching anaesthetists work: using the professional judgement of consultants to assess the developing clinical competence of trainees. AB - One of the responsibilities of a consultant is to identify the clinical competence of trainees by observing them at work. The attributes of clinical competence in anaesthesia were defined by interviewing a group of consultants and trainees. Observation of practice was believed to be important in monitoring competence and the supervising consultants all believed that they could recognize competent performance. The account of the structure of observation provided by the anaesthetists is used as the basis of advice on how to conduct an assessment of trainees' developing professionalism by use of the specialist's professional judgement. PMID- 10823112 TI - Volatile anaesthetics and the atmosphere: atmospheric lifetimes and atmospheric effects of halothane, enflurane, isoflurane, desflurane and sevoflurane. PMID- 10823113 TI - Oxygenator exhaust capnography as an index of arterial carbon dioxide tension during cardiopulmonary bypass using a membrane oxygenator. PMID- 10823114 TI - Perioperative hypotension following plasma volume expansion with albumin in an angiotensin-converting enzyme inhibited infant. PMID- 10823115 TI - Tracheal intubation via the intubating laryngeal mask by inexperienced personnel. PMID- 10823116 TI - Emergency repair of incidentally diagnosed ascending aortic aneurysm immediately after caesarean section. PMID- 10823117 TI - Myocardial ischaemia during caesarean section. PMID- 10823118 TI - Quality control for the immunohistochemical demonstration of oestrogen and progesterone receptors. PMID- 10823120 TI - ACP Best Practice No 158. Neuropathology. PMID- 10823119 TI - The role of EBV in post-transplant malignancies: a review. PMID- 10823121 TI - Monitoring patients with HIV disease. PMID- 10823122 TI - Postmortem findings after fatal anaphylactic reactions. AB - AIMS: To determine the frequency at which classic manifestations of anaphylaxis are present at necropsy after fatal anaphylactic reactions. METHODS: A register has been established of fatal anaphylactic reactions in the UK since 1992, traced from the certified cause of death and other sources. Details of the previous medical history and the reaction suggest anaphylaxis as the cause of death for 130 cases; a postmortem report was available for 56. RESULTS: The 56 deaths studied included 19 reactions to bee or wasp venom, 16 to foods, and 21 to drugs or contrast media. Death occurred within one hour of anaphylaxis in 39 cases. Macroscopic findings included signs of asthma (mucous plugging and/or hyper inflated lungs) (15 of 56), petechial haemorrhages (10 of 56), pharyngeal/laryngeal oedema (23 of 56), but for 23 of 56 there was nothing indicative of an allergic death. Mast cell tryptase was raised in 14 of 16 cases tested; three of three tested had detectable IgE specific for the suspected allergen. CONCLUSIONS: In many cases of fatal anaphylaxis no specific macroscopic findings are present at postmortem examination. This reflects the rapidity and mode of death, which is often the result of shock rather than asphyxia. Investigations that might help determine whether anaphylaxis was the cause of death had rarely been performed. In the presence of a typical clinical history, absence of postmortem findings does not exclude the diagnosis of anaphylaxis. PMID- 10823123 TI - Methadone deaths: a toxicological analysis. AB - AIMS: To perform a toxicological analysis of deaths involving methadone and to determine the fatal concentration of methadone in such deaths. METHODS: Deaths in which methadone was mentioned in the cause of death were identified. Deaths were divided into those associated with methadone only and deaths in which the cause of death was a combination of methadone and other drugs. Toxicological findings in these deaths were analysed and compared with previously published data. RESULTS: One hundred and eleven cases were analysed. In 55 cases, methadone poisoning was given as the sole cause of death. Fifty victims were adults, age range 17-51 years (median, 23), with five victims under 14 years of age. The mean methadone concentration in the adult deaths was 584 micrograms/litre (median, 435; range, 84-2700). In 56 cases, age range 15-49 years, (median, 28), death was ascribed to a combination of methadone and other drugs. The mean methadone concentration in these deaths was 576 micrograms/litre (median, 294; range, 49 2440). In 26 cases, multiple site sampling was performed. This revealed that there could be a 100% discrepancy between methadone concentrations, and other drugs, in samples collected in different sites in the same body. CONCLUSIONS: There is an overlap between quoted therapeutic methadone concentrations and methadone concentrations seen in fatalities. However, those dying from methadone poisoning might not be the same as those in a methadone programme. A degree of caution must be exercised in determining a fatal concentration because of the phenomenon of postmortem redistribution. Pathologists and toxicologists need to examine all the available postmortem findings in identifying the cause of death. PMID- 10823124 TI - Estimating antemortem drug concentrations from postmortem blood samples: the influence of postmortem redistribution. AB - AIMS: To compare blood drug concentrations during life with postmortem drug concentrations measured from a peripheral site and a central site. METHODS: Coroner's cases from October 1990 to July 1997 were reviewed. Six cases had data on both antemortem and postmortem blood drug concentrations. The postmortem to antemortem ratio was compared with the postmortem central to peripheral ratio, using cardiac blood as a central site and femoral blood as a peripheral site. RESULTS: Drugs that have a high postmortem central to peripheral ratio; that is, drugs that exhibit considerable postmortem redistribution, also have high postmortem to antemortem ratios. CONCLUSIONS: A large degree of error can arise from attempting to estimate antemortem drug concentrations and the ingested dose from postmortem measurements. The chosen site and technique for postmortem blood sampling can greatly influence the concentration of drug measured. PMID- 10823125 TI - HLA-DR and beta 2 microglobulin expression in medullary and atypical medullary carcinoma of the breast: histopathologically similar but biologically distinct entities. AB - AIMS: To examine the expression of HLA-DR and beta 2 microglobulin in medullary carcinoma and atypical medullary carcinoma of the breast to determine if the effective presentation of tumour antigens to the immune system can differentiate between these two histopathologically similar entities. METHODS: Expression of HLA-DR and beta 2 microglobulin was examined by immunohistochemical methods in five samples of medullary carcinoma of the breast, which has a relatively favourable prognosis, six samples of atypical medullary carcinoma of the breast, which has a prognosis closer to that of regular invasive duct carcinoma, and 20 samples of invasive duct carcinomas, 10 with an accompanying lymphocytic infiltrate. RESULTS: A positive and significant correlation was found between tumour type and both HLA-DR and beta 2 microglobulin expression. Expression was most prominent in medullary carcinoma, followed by atypical medullary carcinoma and invasive duct carcinoma with and without lymphocytic infiltrates. The mean intensity and percentage of HLA-DR tumour immunostaining were significantly higher in medullary carcinoma than in the other three tumour groups, as was the mean intensity of beta 2 microglobulin immunostaining. Mean percentage of beta 2 microglobulin immunostaining was significantly higher in medullary carcinoma than in invasive duct carcinoma without lymphocytic infiltrates, and showed a trend to increase from invasive duct carcinoma with lymphocytic infiltrates to atypical medullary carcinoma and medullary carcinoma. CONCLUSIONS: Medullary carcinoma and atypical medullary carcinoma of the breast differ in their expression of HLA-DR and beta 2 microglobulin. The relatively favourable prognosis of medullary carcinoma of the breast may be related to effective tumour antigen presentation to the immune system through MHC-I and MHC-II expression. Immunotherapy aimed at MHC-I and MHC-II induction might have a beneficial effect in breast cancer. PMID- 10823126 TI - Immunohistochemical demonstration of oestrogen and progesterone receptors: correlation of standards achieved on in house tumours with that achieved on external quality assessment material in over 150 laboratories from 26 countries. AB - AIMS: To investigate the sensitivity of immunohistochemical (IHC) assays for oestrogen receptors (ER) and progesterone receptors (PR) achieved by laboratories on breast tumours fixed and processed in their own department, and to compare this with the degree of sensitivity they achieve on tumours circulated as part of an external quality assessment (EQA) programme. METHODS: On 10 occasions between April 1994 and June 1998, histological sections from breast cancers showing various degrees of expression of ER and PR were circulated for IHC staining to laboratories participating in the UK national external quality assessment scheme for immunocytochemistry (UK NEQAS-ICC). The staining of these tumours, in addition to that of tumours fixed and processed in the participants own laboratories (in house tumours), was assessed by a panel of four assessors, using the established UK NEQAS-ICC scoring system. For a selected assessment run, the degree of expression of participants in house tumours was evaluated by means of the semiquantitative quick score method. RESULTS: Although the scores awarded for the staining of in house tumours were generally higher than those awarded for the staining of UK NEQAS tumours, there was also a significant positive correlation between the two sets of scores. Using the quick score method of evaluation for one of the assessment runs, 47% of in house tumours were classified as having a high degree of ER expression. Of the remaining cases, a significant proportion initially classified as having only low or medium expression of ER were found to have higher expression when stained by the organising laboratory. The UK NEQAS ICC centre's routine assay for hormonal receptors was found to be 90-100% efficient in achieving optimal demonstration of breast tumours from over 150 different laboratories. CONCLUSIONS: The significant positive correlation between the results obtained on the UK NEQAS tumours and the in house tumours provides evidence for the view that results achieved on EQA material are accurate indicators of in house laboratory performance. Although most laboratories adequately detected tumours with high receptor expression, a large proportion of in house tumours classified initially by participants' staining as being of low or medium ER expression had a higher degree of expression when stained by the UK NEQAS-ICC centre. The efficiency of the organising centre's routine IHC method for ER and PR in optimally demonstrating participants in house breast tumours shows that variations in fixation and tissue preparation are not limiting factors preventing a different laboratory achieving optimal demonstration. PMID- 10823127 TI - Dietary calcium supplementation increases apoptosis in the distal murine colonic epithelium. AB - BACKGROUND: Increased dietary calcium might reduce colorectal cancer risk, possibly by reduction of colonic epithelial hyperproliferation, but not all studies have demonstrated this. Little is known about the effects of calcium on colonic apoptosis. AIM: To quantify the effects of increasing calcium on apoptosis and cell proliferation in normal murine colonic crypt epithelium. METHODS: Twenty one day old male C57B1/6 mice were fed either control AIN-76 diet (0.5% calcium wt/wt; n = 10) or the same supplemented with calcium carbonate (1.0% calcium; n = 10) for 12 weeks. Apoptotic cells in proximal and distal segments were counted and expressed as an apoptotic index (AI: frequency of apoptosis/100 longitudinal crypts). The bromodeoxyuridine (BrdU) labelling index was also determined. Differences were analysed by the student's t test. RESULTS: In control animals, the AI was significantly higher in the caecum/proximal colon (mean, 28.6; SEM, 2.0) compared with the distal colon (mean, 19.9; SEM, 1.8; p = 0.004). In the calcium treated group, the AI in the caecum/proximal colon (mean, 30.6; SEM, 1.7) was similar to controls (p = 0.71) but the AI in the distal colon was significantly greater (mean, 32.6; SEM, 1.8; p = 0.001) than in control mice and was raised to values similar to those in the proximal colon. Calcium was also associated with reduced crypt cellularity and, in the proximal colon, a downward shift in the crypt position at which apoptosis occurred. There were no significant differences in the BrdU labelling index between groups or between proximal and distal colonic segments in each group. CONCLUSIONS: Increased dietary calcium is associated with the induction of apoptosis in normal mouse distal colonic epithelium without affecting cell proliferation. This might contribute to its putative chemopreventive role in colorectal carcinogenesis. Whether this effect is direct or indirect requires further study. PMID- 10823128 TI - Wet mount microscopy reflects functional vaginal lactobacillary flora better than Gram stain. AB - AIM: The status of vaginal lacto-bacillary flora, an indicator of possible genital infection and pregnancy complications, can be assessed on wet mount or Gram stained specimens. The former is quick, the latter more routine. The accuracy of the two preparative techniques to detect normal vaginal lacto bacillary microflora was compared for 646 patients. The effect of delay in transport medium before Gram staining was also investigated. METHODS: Patients presented with infectious vaginitis or for a routine prenatal visit. After placement of a speculum, duplicate smears were taken from the upper vaginal vault and examined fresh or after Gram staining. Lacto-bacillary grades from both methods were compared with lactate concentration in vaginal rinses. In a subgroup of 238 patients, Gram staining was performed both on fresh smears and those that had been transported in Stuart's growth medium. RESULTS: Higher lacto-bacillary grades (more disrupted flora) were diagnosed 2.9 times more frequently on Gram stained specimens than on wet mounts (p < 0.0001), a difference even more pronounced after transport in Stuart's medium (relative risk, 4.2; p < 0.0001). Lacto-bacillary grades assessed on wet mounts correlated better with vaginal lactate concentration than those assessed on Gram stains. CONCLUSIONS: Easier recognition of lacto-bacillary morphotypes on wet mounts than on Gram stains might result from the loss of lactobacilli by the process of fixation or Gram staining. Wet mount microscopy of vaginal smears for assessment of lacto bacillary grades, rather than Gram staining, is strongly recommended. PMID- 10823129 TI - Western blotting is useful in the salivary diagnosis of Helicobacter pylori infection. AB - BACKGROUND: The salivary diagnosis of Helicobacter pylori infection offers attractive possibilities for the epidemiological study of infection in children. Salivary enzyme linked immunosorbent assay (ELISA) is less reliable then serum ELISA, owing to variable transudation of immunoglobulin. In addition, children are more difficult to study because of lower specific serum antibody concentrations to H pylori. The performance of salivary western blotting in comparison with serum western blotting and serum ELISA was investigated in school children. SUBJECTS AND METHODS: Paired serum and saliva specimens were obtained from 669 [corrected] school children aged 9-11 in 10 British towns. All saliva and serum specimens were first analysed by ELISA; subsequently, western blotting of both specimens was performed on 31 and 34 specimens, respectively, to establish the criteria for positivity for western blotting. The remaining 121 specimens were then tested blindly and saliva was compared with the serum. RESULTS: The sensitivity and specificity of salivary ELISA in the 669 [corrected] specimens was 32 of 50 (64%) and 530 of 619 (86%) [corrected], respectively, when compared with serum ELISA. The western blotting validation was performed on 28 subjects with positive serum and positive salivary ELISA, 28 saliva positives with negative serum, 16 saliva negatives with positive serum, and 50 doubly negative subjects. Compared with serum western blots, the sensitivity and specificity of salivary western blots was 38 of 47 (81%) and 68 of 75 (91%), respectively. Using serum ELISA as the gold standard, the sensitivity and specificity were 32 of 44 (73%) and 72 of 78 (92%), respectively, the specificity being significantly higher than salivary ELISA (p < 0.001). CONCLUSION: Salivary western blotting for IgG is useful in the diagnosis of H pylori infection and is superior to ELISA. It also permits the identification of pathogenic strains. PMID- 10823130 TI - Co-expression in Helicobacter pylori of cagA and non-opsonic neutrophil activation enhances the association with peptic ulcer disease. AB - AIMS: To investigate the association of cagA positivity and non-opsonic neutrophil activation capacity in wild-type Helicobacter pylori strains with peptic ulcer disease or chronic gastritis only. METHODS: Helicobacter pylori were isolated from antral biopsies of 53 consecutive patients with chronic antral gastritis, of whom 24 had peptic ulcer disease endoscopically. The presence of cagA, a marker for the cag pathogenicity island, was determined by polymerase chain reaction with specific oligonucleotide primers, and non-opsonic neutrophil activation capacity by luminol enhanced chemiluminescence. RESULTS: The cagA gene was present in 39 of 53 (73.6%) strains, 20 of which (83.3%) were from the 24 patients with peptic ulcer disease and 19 (65.5%) from the 29 patients with chronic gastritis only. Non-opsonic neutrophil activation was found in 29 (54.7%) strains, 16 of which (66.7%) were from patients with peptic ulcer disease, and 13 (44.8%) from those with chronic gastritis. Non-opsonic neutrophil activation was found more frequently in cagA+ than cagA- strains (59% v 42.9%). Whereas four of the 14 cagA- strains and eight of the 24 non-opsonic neutrophil activation negative strains were from patients with peptic ulcer disease, only two of 24 (8.3%) peptic ulcer disease strains expressed neither cagA nor non-opsonic neutrophil activation. The cagA gene and non-opsonic neutrophil activation capacity were co-expressed in 14 of 24 (58.3%) strains from patients with peptic ulcer disease, and in nine of 29 (31%) strains from individuals with chronic gastritis. CONCLUSIONS: Positivity for cagA and non-opsonic neutrophil activation occur independently in wild-type H pylori strains. However, co-expression of the two markers enhanced the prediction of peptic ulcer disease. PMID- 10823131 TI - Effect of transforming growth factor beta 2 on oestrogen metabolism in the MCF-7 breast cancer cell line. AB - Transforming growth factor beta (TGF-beta) is a multifunctional regulator of cellular growth and differentiation in many cell types and has a growth inhibitory effect on mammary epithelial cells. The TGF-beta 2 isoform has been shown to be present in high concentrations in breast cyst fluid and might have a protective role in breast cancer. In addition, oestrogens play an important role in breast cancer development, and oestrone sulphate (E1S) might be the main source of active oestrogens in the breast. The aim of this study was to assess the effect of TGF-beta 2 on oestrogen synthesis in an attempt to understand the mechanism by which TGF-beta 2 may exert a protective effect in breast cancer. In this study, higher concentrations of TGF-beta 2 significantly inhibited the conversion of E1S to oestrone (E1) and the conversion of E1 to the potent oestrogen, oestradiol (E2). TGF-beta 2 did not have any effect on MCF-7 cell growth or on E2 to E1 conversion. In conclusion, TGF-beta 2 might exert a protective role in breast cancer by reducing the amount of active oestrogens present in the breast. PMID- 10823132 TI - Thrombophilia testing: what do we think the tests mean and what should we do with the results? PMID- 10823133 TI - ACP Best Practice No 157. Guidelines for the laboratory handling of laryngectomy specimens. PMID- 10823134 TI - Benign monoclonal expansion of CD8+ lymphocytes in HIV infection. AB - BACKGROUND: A transient expansion of the CD8+ T cell pool normally occurs in the early phase of HIV infection. Persistent expansion of this pool is observed in two related settings: diffuse infiltrative lymphocytosis syndrome (DILS) and HIV associated CD8+ lymphocytosis syndrome. AIM: To investigate a group of HIV infected patients with CD8+ lymphocytosis syndrome with particular emphasis on whether monoclonality was present. METHODS: A group of 18 patients with HIV-1 infection and persistent circulating CD8+ lymphocytosis was compared with 21 HIV positive controls. Serum samples were tested for antinuclear antibodies, antibodies to extractable nuclear antigens, immunoglobulin levels, paraproteins, human T lymphotropic virus type 1 (HTLV-1), Epstein-Barr virus, and cytomegalovirus serology. Lymphocyte phenotyping and HLA-DR typing was performed, and T cell receptor (TCR) gene rearrangement studies used to identify monoclonal populations of T cells. CD4+ and CD8+ subsets of peripheral blood lymphocytes were purified to determine whether CD8+ populations inhibited HIV replication in autologous CD4+ cells. RESULTS: A subgroup of patients with HIV-1 infection was found to have expanded populations of CD8+ T cell large granular lymphocytes persisting for 6 to 30 months. The consensus immunophenotype was CD4- CD8+ DRhigh CD11a+ CD11c+ CD16- CD28+/- CD56- CD57+, consistent with typical T cell large granular lymphocytes expressing cellular activation markers. Despite the finding of monoclonal TCR gene usage in five of 18 patients, there is evidence that the CD8+ expansions are reactive populations capable of mediating non-cytotoxic inhibition of HIV replication. CONCLUSIONS: A subgroup of HIV positive patients has CD8+ lymphocytosis, but despite the frequent occurrence of monoclonal TCR gene usage there is evidence that this represents an immune response to viral infection rather than a malignant disorder. PMID- 10823135 TI - Improved prognosis of Epstein-Barr virus associated childhood Hodgkin's lymphoma: study of 47 South African cases. AB - AIM: To study the distribution of Hodgkin's lymphoma in South African children and report the incidence of Epstein-Barr virus (EBV) as regards age, race, sex, and histological subtype; to investigate whether EBV is relevant to survival. METHODS: Immunohistochemistry (IHC) and in situ hybridisation (ISH) to detect EBV were performed on 47 South African children with classical Hodgkin's lymphoma, ranging in age from 3 to 14 years and coming from different ethnic backgrounds. The correlation between the presence of the virus and clinical outcome was assessed. RESULTS: The nodular sclerosing subtype predominated, comprising 89% of cases; the remaining 11% were of the mixed cellularity subtype. EBV was present in 68%. Full clinical data were available for 36 cases; EBV positive patients presented with less aggressive symptoms at diagnosis and had a significantly longer median survival than EBV negative patients. CONCLUSIONS: The distribution of EBV in South African childhood Hodgkin's lymphoma follows a pattern intermediate to that of industrialised and non-industrialized countries. Furthermore, our data suggest that there is an association between poor prognosis and the non-detection of EBV products in South African childhood Hodgkin's lymphoma. PMID- 10823136 TI - Clinicopathological features of gastric mucosa associated lymphoid tissue (MALT) lymphomas: high grade transformation and comparison with diffuse large B cell lymphomas without MALT lymphoma features. AB - AIMS: To investigate the clinicopathological differences among gastric low grade MALT lymphomas (low MALT), large B cell lymphomas with low grade components (secondary high grade MALT lymphomas, high MALT), and diffuse large B cell lymphomas without low grade features (primary high grade MALT lymphomas, DLL). METHODS: Clinicopathological and morphological characters of 126 gastric lymphoma cases were studied: 82 cases of low MALT lymphoma including 40 that were surgically resected, 17 cases of high MALT lymphoma including 13 surgically resected, and 27 cases of DLL including 12 surgically resected. RESULTS: Age ranges were as follows: low MALT lymphoma, 34 to 85 years (mean 59.9); high MALT lymphoma, 53 to 88 years (mean 68.5); DLL, 29 to 83 years (mean 62.3). The average age for low and high MALT lymphomas was significantly different (p < 0.05), but there were no differences in other comparisons. There was a female predominance of low MALT lymphoma patients (female to male ratio, 47/35), while for high MALT patients the ratio was almost even (8/9), and for DLL patients there was a male predominance (11/16). Examination of surgically resected material showed that MALT lymphomas had a wider distribution in the gastric wall than DLL. CONCLUSIONS: The findings suggest that at least some of the high grade gastric lymphomas, especially in patients younger than the fifth decade, do not originate from high grade transformation of low MALT lymphomas. It seems to take about one decade at least for high grade transformation of low MALT lymphomas. PMID- 10823137 TI - Completeness of excision and follow up cytology in patients treated with loop excision biopsy. AB - AIMS: To assess the relation between the grade and the status of follow up cytology, the completeness of loop excision biopsies with cervical intraepithelial neoplasia (CIN), and the findings at follow up cytology, as well as the differences between complete and incomplete exclusion, using the odds ratio. Treatment failure was assessed. METHODS: 1600 women with CIN (290 CIN1, 304 CIN2, 1006 CIN3) were followed for a minimum of six months and a maximum of 10 years. A database was created and comparisons performed. The mean age of the patients was 37 years. RESULTS: Excision was complete in over 84% of loops. Residual disease and recurrence of high grade dyskaryosis was more common in women with CIN 3 than CIN 2 or 1. No high grade dyskaryosis was seen in the fifth follow up smear in patients with CIN 1 and CIN 2. Residual, recurrent, and persistent disease was most common in patients with incompletely excised CIN at ectocervical and endocervical margins and deep margins of resection than in patients with completely excised CIN. The odds ratios were significantly higher in the women who had incomplete excision of CIN at ectocervical, endocervical, both ecto- and endocervical, and deep margins of resection compared with those with apparent complete excision of CIN lesions. One patient developed invasive squamous cell carcinoma 44 months after loop excision which showed CIN 3 invading endocervical crypts and extending to both ectocervical and endocervical margins of resection. CONCLUSIONS: At long term follow up, patients with CIN who have residual disease are at increased risk of persistent disease and should therefore be followed up regularly with cytology and colposcopy. The findings support national policy of returning women with treated CIN of any grade to normal recall after five years except for cases of CIN3 where excision was incomplete or equivocal. In these cases follow up with annual smear for 10 years is recommended. PMID- 10823138 TI - Apoptosis in cervical squamous carcinoma: predictive value for survival following radiotherapy. AB - BACKGROUND: Apoptosis, or programmed cell death, can be induced by radiotherapy. The extent of apoptosis in a tumour before treatment may have important implications for response to radiotherapy and long term survival. AIM: To examine the extent of apoptosis in tumour tissue from patients with squamous carcinoma of the cervix before radiotherapy, and to correlate this with response to treatment and prognosis. METHODS: The percentage of apoptotic cells was assessed in 146 carcinomas of the cervix from patients scheduled to receive radiotherapy. The CAS 200 static image analysis system was used to count the number of tumour nuclei per high power field, while the numbers of apoptotic cells in the same field were visualised simultaneously on the image analyser and recorded manually. RESULTS: The median apoptotic level was 0.73%. Patients were divided into two groups around the median. There was no statistically significant difference in outcome between the two groups as determined by long term survival following radiotherapy. CONCLUSIONS: The CAS 200 static image analyser system can be used to assist in the rapid semiautomated assessment of apoptosis in conventionally prepared tissue. The results suggest that the apoptotic state of a tumour before treatment is of no value in predicting response to radiotherapy and subsequent prognosis. Tumour stage, size, and BrdU labelling index, as a measure of proliferation rate, remain the most important prognostic factors in terms of predicting local tumour control. PMID- 10823139 TI - Oestrogen, progesterone, and androgen receptors in ovarian neoplasia: correlation between immunohistochemical and biochemical receptor analyses. AB - AIM: To investigate the correlation between immunohistochemical and biochemical steroid receptor analyses by measurement of oestrogen, progesterone, and androgen receptor status in ovarian neoplasia. METHODS: Tissue samples were obtained from 27 ovarian neoplasms, including two borderline tumours. Immunohistochemical staining of the tissue slides was scored semiquantitatively, incorporating the intensity and percentage of positive staining (histo-score). Tumours with a histo score of 10 or more were considered steroid receptor positive. The epithelial and stromal fractions of the tumours were analysed separately. To study the uniformity of receptor expression throughout a tumour, up to four samples were analysed. RESULTS: Immunohistochemical histo-scores of the oestrogen receptor in the epithelial fractions were significantly correlated with the biochemical oestrogen receptor values (r = 0.408). Androgen receptor status in the epithelial fraction was correlated with that in the stromal fraction (r = 0.741), while androgen receptor histo-scores in the epithelial fraction correlated with the biochemical assay values (r = 0.463). On biochemical analysis, 17 of the 27 ovarian tumours were oestrogen receptor positive and seven were progesterone receptor positive. On immunohistochemical analysis, eight tumours were oestrogen receptor positive and two were progesterone receptor positive. Biochemical analysis showed that 14 of the 26 tumours were slightly androgen receptor positive (10-50 fmol/mg protein), while all the others were negative. On immunohistochemical analysis, seven of the 26 tumours were androgen receptor positive. When two or more specimens from one tumour were analysed, marked differences in steroid status were found, especially in progesterone receptor and androgen receptor expression. Some parts of a tumour were steroid receptor positive, while other parts were negative owing to heterogeneity of expression. CONCLUSIONS: Immunohistochemical and biochemical analysis of steroid receptors in ovarian tumours correlated weakly or not at all. Heterogeneity of expression within a tumour and the presence of progesterone and androgen receptors in the stromal fraction partly accounted for this observation. Biochemical and immunohistochemical androgen receptor status was much lower than in previous reports. PMID- 10823140 TI - Value of A103 (melan-A) immunostaining in the differential diagnosis of ovarian sex cord stromal tumours. AB - AIMS: To assess A103 (melan-A) immunoreactivity in a range of ovarian sex cord stromal tumours and to evaluate it for the differential diagnosis of other neoplasms. METHODS: Paraffin embedded tissue sections from 45 sex cord stromal tumours and 44 potential histological mimics were examined immunohistochemically using the antibody A103. The sex cord stromal group included 21 adult granulosa cell tumours (AGCT), two juvenile granulosa cell tumours (JGCT), eight tumours showing Sertoli cell or Sertoli-Leydig cell differentiation, two unclassified tumours, two gonadoblastomas, one sex cord tumour with annular tubules, two steroid cell tumours, five thecomas/fibrothecomas, and two sclerosing stromal tumours. The histological mimics include 14 primary ovarian carcinomas, 13 metastatic carcinomas, four carcinoid tumours, four lymphomas, three endometrioid stromal sarcomas, two ovarian tumours of probable Wolffian origin, and one case each of small cell carcinoma, desmoplastic small round cell tumour, melanoma, and primitive neuroectodermal tumour. RESULTS: A103 immunoreactivity was identified in 25 sex cord stromal tumours including 10 AGCT, two JGCT, six Sertoli/Sertoli Leydig cell tumours, two steroid cell tumours, three thecomas/fibrothecomas, and two sclerosing stromal tumours. Of the potential histological mimics, staining was present only in the two ovarian tumours of probable Wolffian origin and the melanoma. Immunoreactive stromal cells were noted in a minority of cases. Normal hilus cells and rete ovarii epithelium also expressed A103. CONCLUSIONS: A103 is a moderately sensitive and specific marker of sex cord stromal differentiation within the range of tumours examined in this study and as such is a valuable adjunct to other immunocytochemical markers in the assessment of diagnostically problematic ovarian tumours. The staining of normal and neoplastic Wolffian elements merits further investigation. PMID- 10823141 TI - Expression of a homeobox gene (SIX5) in borderline ovarian tumours. AB - AIMS: To assess the expression of SIX5 (a homeobox gene) mRNA in surface coelomic epithelium, endocervical epithelium, Fallopian tube epithelium, and benign, borderline, and malignant epithelial ovarian tumours. METHODS: 10 normal premenopausal ovaries, 10 normal Fallopian tubes, 10 normal cervices, 10 normal postmenopausal ovaries, 10 benign epithelial ovarian tumours, 10 malignant epithelial ovarian tumours, and 40 borderline epithelial ovarian tumours were studied retrospectively. The tissues had been fixed in formalin and embedded in paraffin wax. The tumours had previously been typed into mucinous, serous, or mixed tumours and assigned to the borderline category according to the FIGO/WHO criteria. Expression was assessed by in situ binding of SIX5 specific sense and antisense riboprobes. Hybridization of the riboprobes was detected using a standard immunohistochemical technique and the results correlated with expression in the normal epithelium of the endocervix, Fallopian tube, surface coelomic epithelium, and ovarian tumours. RESULTS: Expression of SIX5 mRNA was demonstrated in normal Fallopian tube epithelium and normal endocervical epithelium. SIX5 mRNA was not detected in normal ovarian epithelial tissue at any of the times studied during the menstrual cycle. Expression of SIX5 was not shown in benign epithelial ovarian tumours or in any of the malignant epithelial ovarian tumours. In 31 of 37 borderline epithelial ovarian tumours (84%), SIX5 expression was found in the epithelial cells. CONCLUSIONS: SIX5 expression is present in the normal epithelium throughout most of the female reproductive tract, suggesting it may have a role in maintaining epithelial differentiation in these tissues. SIX5 expression appears to be restricted to borderline epithelial ovarian tumours and may be a marker of epithelial differentiation in these tumours; thus borderline ovarian tumours may not be part of a continuum of disease between benign and malignant epithelial ovarian tumours. Further investigation of expression of SIX5 may clarify the molecular processes that promote differentiation of the ovarian surface epithelium. PMID- 10823142 TI - Quantitation of Helicobacter pylori in dental plaque samples by competitive polymerase chain reaction. AB - AIM: To establish a competitive PCR (cPCR) assay for quantitation of H pylori organisms in dental plaque samples. METHODS: The cPCR co-amplified target H pylori DNA and a known amount of internal standard template in the same tube with the same primers directed to 0.86 kb DNA of H pylori. The internal standard was a synthesised DNA bearing the same primer recognition sites at two ends and a non homologous core sequence as the target DNA fragment. Quantitation was based on determination of the relative, not absolute, amounts of the differently sized and [32P]-dCTP labelled products derived from H pylori DNA and the competitive internal standard after gel electrophoresis separation. RESULTS: A significant correlation between known amounts of H pylori added to dental plaque samples and the results of the cPCR was found, and a standard line was developed which allowed quantitation of H pylori in the plaque samples. cPCR was performed on supragingival plaque samples from 10 adult patients with H pylori infection in the stomach, and from five adults and six children without H pylori infection in the stomach. The ranges of H pylori numbers were 1-213 (median 25), 6-76 (10), and 4-94 (14) cells/mg of dental plaque in the three groups, respectively. CONCLUSIONS: cPCR is useful for quantitation of H pylori in supragingival dental plaque samples; however, the number of the organisms in dental plaque samples seems very low. PMID- 10823143 TI - Muscle changes in the neuroleptic malignant syndrome. AB - AIMS: To characterise the skeletal muscle changes in the neuroleptic malignant syndrome (NMS). METHODS: Detailed light and ultrastructural examination was carried out on skeletal muscle from three cases of NMS, two associated with recreational drugs (3,4-methlenedioxymethylamphetamine (MDMA, Ecstasy) and lysergic acid diethylamide (LSD)) and one with antipsychotic drugs (fluoxetine (Prozac) and remoxipride hydrochloride monohydrate (Roxiam)). RESULTS: The muscles were grossly swollen and oedematous in all cases, in one with such severe local involvement that the diagnosis of sarcoma was considered. On microscopy, there was conspicuous oedema. In some fascicles less than 10% of fibres were affected whereas in others more than 50% were pale and enlarged. There was a spectrum of changes: tiny to large vacuoles replaced most of the sarcoplasm and were associated with necrosis. A striking feature in some fibres was the presence of contraction bands separating segments of oedematous myofibrils. Severe endomysial oedema was also detectable. There was a scanty mononuclear infiltrate but no evidence of regeneration. CONCLUSIONS: The muscle changes associated with NMS are characteristic and may be helpful in differential diagnosis. PMID- 10823144 TI - Splenic B cell lymphoma with lymphocyte clusters in peripheral blood smears. AB - EDTA induced clumping of lymphoid cells, both benign and malignant, in peripheral blood samples has been reported only rarely. Such clustering presents the laboratory and pathologist with unique difficulties in the accurate diagnosis of these disorders. A case of low grade B cell splenic lymphoma is presented with lymphocyte clumping in smears made from EDTA anticoagulated peripheral blood, the fourth case described in which neoplastic lymphoid cells form clusters in vitro in peripheral blood. The association with splenic lymphomas (three of the four cases) is intriguing but its significance remains uncertain. PMID- 10823145 TI - Clinical value of anaerobic blood culture: a retrospective analysis of positive patient episodes. AB - AIM: To investigate the clinical value of anaerobic blood culture. METHODS: Blood culture bottles (n = 25,185) submitted for culture over a two year period were reviewed. RESULTS: The bottles yielded 1992 positive patient episodes, a positive rate of 14.4/1000 hospital admissions. Significantly more isolations were obtained from aerobic than from anaerobic bottles. Twelve of the 38 anaerobic episodes were detected in aerobic bottles. Clinical management was influenced in one of 24 patients whose cultures yielded anaerobes from anaerobic bottles only. For a further six patients it was unlikely that the result had any effect on clinical management. CONCLUSIONS: If aerobic bottles were substituted for the anaerobic bottles, detection of positive patient episodes would increase by at least 6%. A higher yield would be achieved by using two aerobic bottles for routine culture and using anaerobic bottles only for patients where anaerobic culture may influence clinical management. PMID- 10823146 TI - Pasteurella multocida meningitis in an adult: case report. AB - Pasteurella multocida is known to form part of the normal flora in the nasopharynx or gastrointestinal tract in many domestic and wild animals. Most human P multocida infections are soft tissue infections caused by dog or cat bites. Less commonly this bacterium is associated with infections affecting other organ systems of man. A case of fatal P multocida meningitis discovered at the necropsy of a 52 year old man is described. P multocida is an unusual causative agent of meningitis which tends to affect those at the extremes of age. PMID- 10823147 TI - Type 1 diabetes mellitus masking primary antibody deficiency. AB - A patient with a history of recurrent cutaneous and pulmonary infections, nephrotic syndrome, and an established diagnosis of type 1 diabetes was found to have unsuspected and unrecognised primary immunodeficiency. On review of the case, previous investigations pointed to the correct diagnosis over 10 years earlier. This combination of diagnoses has not previously been reported. The patient is now well on replacement intravenous immunoglobulin therapy, urinary loss of IgG having been specifically excluded before treatment. This case highlights how antibody deficiency can easily be missed despite an obvious infection history unless results are interpreted carefully and in context. PMID- 10823148 TI - Application of microsatellite PCR techniques in the identification of mixed up tissue specimens in surgical pathology. AB - A fragment of tumour was erroneously mixed up with an endometrial biopsy. Micro satellite polymerase chain reaction (PCR) clearly demonstrated the extraneous nature of the fragment. Micro-satellite PCR may be useful for the identification of mis-labelled or mismatched tissue fragments in surgical pathology specimens. PMID- 10823149 TI - Pregnancy associated endometriosis with pronounced stromal myxoid change. AB - A case of endometriosis presenting as a mass in the groin of a pregnant woman is described. The mass increased in size during the pregnancy and the radiological features were suspicious of malignancy. Histological examination showed atrophic glands set in an abundant stroma. This was not typical of normal endometrial stroma but had a pronounced myxoid appearance with areas of decidualization. The atypical site of the endometriosis together with the unusual stromal changes resulted in diagnostic confusion. Although stromal decidualization is well recognized in endometriosis in pregnancy, pronounced myxoid change appears unusual. The myxoid change in this case may be a degenerative phenomenon related to pregnancy. PMID- 10823150 TI - Necropsy organ weights are largely useless. PMID- 10823151 TI - Here we go again. Organized medicine launches an attack on nursing. PMID- 10823152 TI - Out from under their thumbs. Who's to blame when managers abuse their power? PMID- 10823153 TI - Family presence research strikes a chord. PMID- 10823154 TI - Family presence research strikes a chord. PMID- 10823155 TI - Family presence research strikes a chord. PMID- 10823156 TI - Stop labeling patients. PMID- 10823157 TI - A conscious sedation resource. PMID- 10823159 TI - Maine's Medicaid demonstration plan now covers early treatment for people who are HIV positive. PMID- 10823158 TI - 80% of car seats unsafe. PMID- 10823160 TI - JCAHO standards in long-term care. PMID- 10823161 TI - A rose by any other name may not smell as sweet. Don't rely on name recognition. PMID- 10823162 TI - Uniformity. Why go back to wearing white? PMID- 10823163 TI - Who's afraid of hepatitis C? PMID- 10823164 TI - Counting nurses. PMID- 10823165 TI - Surveying newly licensed nurses in New York State. PMID- 10823166 TI - The promise of recombinant interleukin-2. PMID- 10823167 TI - Emergency. Delirium tremens. PMID- 10823168 TI - Are we starving our patients? PMID- 10823169 TI - Hastening death. PMID- 10823170 TI - One drink too many. Is there no safe level of alcohol consumption during pregnancy? PMID- 10823171 TI - Protect yourself from sexual harassment. PMID- 10823172 TI - The value of patient- and family-centered care. PMID- 10823173 TI - Presidential endorsement process begins. PMID- 10823174 TI - Guarding against radiation exposure. PMID- 10823175 TI - A clinical advancement program: evaluating 10 years of progressive change. AB - OBJECTIVE: To report the evolution of a clinical advancement program, UEXCEL, at a western teaching hospital and the outcomes associated with evaluation over time. BACKGROUND: The clinical ladder program was initiated in 1989 to provide a professional framework for developing, evaluating, and promoting registered nurses. The program is derived from Benner's Novice to Expert model. Over a 10 year period, the program has undergone three significant revisions. Program evaluation data have been used to guide institutional change. METHODS: Structure and progression of program development and change are outlined. Evaluation data were collected using a 23-item clinical ladder satisfaction scale developed by Strzelecki. Data were collected in 1993, 1994, 1996, and 1998 using standard survey methods after institutional review board approval. Subjects were registered nurses holding clinical positions at the University of Colorado Hospital. Data were trended across units and time periods and were compared with other institutional evaluation data sets. RESULTS: Improvement in nurse satisfaction with the UEXCEL program has been steady and incremental, after low baseline measurement. Satisfaction has improved after each program revision. A significant demographic variable over time is the correlation between higher registered nurse education and program satisfaction. Human resources issues are reported with data results. CONCLUSIONS: Sustaining a clinical advancement program represents a challenge in the current health-care environment. Institutional commitment, staff involvement in revisions, and activities to improve professional nurse development are critical strategies so progress can be achieved. PMID- 10823176 TI - Hospital ownership, performance, and outcomes: assessing the state-of-the science. AB - OBJECTIVE: This study 1) identified the research evidence; 2) assessed the state of-the-science surrounding hospital ownership, performance, and outcomes in acute care hospitals in the United States; and 3) identified measurable components of hospital performance and outcomes for the organization, patient, and community. BACKGROUND: As the size of the nonprofit sector decreases and the size of the for profit sector increases, hospital ownership warrants examination. Most research has focused on either ownership and performance or ownership and outcomes, rather than the potential interaction of all three variables. METHODS: A comprehensive, computerized search of the healthcare research literature yielded 69 data-based references published between 1985 and 1999. Coding sheets were developed to abstract the articles. Analysis involved synthesizing the research evidence for each of the three major variables and their components. RESULTS: Hospital ownership has an impact on hospital performance in relation to system operations; costs, prices, and financial management practices; and personnel issues. Organizational outcomes are similar among hospital ownership types in relation to increasing administrative costs and overall mediocre efficiency. Organizational outcomes differ among hospital ownership types in relation to nursing staff mix and professional satisfaction. The association of hospital ownership with patient outcomes varies depending on the dimension measured. The evidence is mixed or inconclusive regarding hospital ownership and access to care, morbidity, and mortality. The association of hospital ownership and adverse events is consistently supported. Hospital ownership status has an impact on the type and magnitude of community benefits. Differences among the three hospital ownership types are minimized in a competitive market. CONCLUSIONS: This study reinforces the position that nurse researchers need to include hospital ownership as an important structural variable in their studies of hospital-based nursing. Examining the conceptual links between ownership, performance, and outcomes requires the integration of macro-level and micro-level theory. PMID- 10823177 TI - Nurse executive transformational leadership found in participative organizations. AB - OBJECTIVE: The study examined a national sample of 396 randomly selected hospital nurse executives to explore transformational leadership, stage of power, and organizational climate. BACKGROUND: Results from a few nurse executive studies have found nurse executives were transformational leaders. As executives were more transformational, they achieved better staff satisfaction and higher work group effectiveness. This study integrates Bass' transformational leadership model with Hagberg's power stage theory and Likert's organizational climate theory. METHODS: Nurse executives (396) and staff reporting to them (1,115) rated the nurse executives' leadership style, staff extra effort, staff satisfaction, and work group effectiveness using Bass and Avolio's Multifactor Leadership Questionnaire. Executives' bosses (360) rated executive work group effectiveness. Executives completed Hagberg's Personal Power Profile and ranked their organizational climate using Likert's Profile of Organizational Characteristics. RESULTS: Nurse executives used transformational leadership fairly often; achieved fairly satisfied staff levels; were very effective according to bosses; were most likely at stage 3 (power by achievement) or stage 4 (power by reflection); and rated their hospital as a Likert System 3 Consultative Organization. Staff satisfaction and work group effectiveness decreased as nurse executives were more transactional. Higher transformational scores tended to occur with higher educational degrees and within more participative organizations. CONCLUSIONS: Transformational qualities can be enhanced by further education, by achieving higher power stages, and by being within more participative organizations. PMID- 10823178 TI - Evaluating nursing administration instruments. AB - OBJECTIVE: To identify and evaluate available measures that can be used to examine the effects of management innovations in five important areas: autonomy, conflict, job satisfaction, leadership, and organizational climate. BACKGROUND: Management interventions target the context in which care is delivered and through which evidence for practice diffuses. These innovations need to be evaluated for their effects on desired outcomes. However, busy nurses may not have the time to locate, evaluate, and select instruments to measure expected nursing administration outcomes without research-based guidance. Multiple and complex important contextual variables need psychometrically sound and easy-to use measurement instruments identified for use in both practice and research. METHOD: An expert focus group consensus methodology was used in this evaluation research to review available instruments in the five areas and evaluate which of these instruments are psychometrically sound and easy to use in the practice setting. RESULTS: The result is a portfolio of measures, clustered by concept and displayed on a spreadsheet. Retrieval information is provided. The portfolio includes the expert consensus judgment as well as useful descriptive information. CONCLUSIONS: The research reported here identifies psychometrically sound and easy-to-use instruments for measuring five key variables to be included in a portfolio. The results of this study can be used as a beginning for saving time in instrument selection and as an aid for determining the best instrument for measuring outcomes from a clinical or management intervention. PMID- 10823179 TI - Knowledge and attitudes of home care nurses toward hospice referral. AB - OBJECTIVE: Nurses and administrators are concerned with removing barriers to appropriate and cost-effective referrals between programs. The goal of this research was to describe knowledge and attitudes of home care nurses toward hospice referral. BACKGROUND: The literature review did not identify studies related to the referral of patients from traditional home care to hospice programs. A few studies related to the hospice referral processes suggested issues with conflicts between programs, attitude barriers to open communications, issues regarding late hospice referrals, and practitioner reluctance to "give up" patients with whom they had established relationships. METHOD: This study used a descriptive, quantitative method. A 15-item, self-administered survey was developed by the investigators and hospice administrators. It was distributed to 160 registered nurses employed as full- or part-time staff in a large midwestern home care agency. Completed surveys were returned by 75 nurses, for a response rate of 46.9% (N = 75). RESULTS: Nurses perceived that home care and hospice services were very similar; they resisted hospice referral as long as they felt that services could be provided adequately by home care; they desired to maintain patient continuity and special rapport and demonstrated inconsistent knowledge regarding hospice referral criteria and relative costs. CONCLUSIONS: Nurses in different programs would benefit from improved communication links, accurate cross-program information, and development of increased trust relationships. Improvements in communication and transition programs for patients and families should include structural and programmatic changes. PMID- 10823180 TI - Registered nurses, unlicensed assistive personnel, and organizational culture in hospitals. AB - OBJECTIVE: This study investigated "thinking and behavioral" styles that are used to measure the concept of organizational culture among registered nurses (RNs) and unlicensed assistive personnel (UAPs) in acute care hospitals. BACKGROUND: The Organizational Culture Inventory has been used in numerous industries, including hospitals, to describe aspects of organizational culture in order to better understand and manage change. Evidence shows that nursing unit and hospital culture tends to be "constructive" but little evidence indicates whether workers in different job categories influence the culture differently. METHOD: This survey was conducted in five tertiary care hospitals on the west coast of the United States. The design is cross-sectional and analytic. A convenience sample was recruited from staff members working at least 20 hours per week in the adult medical-surgical and specialty nursing units. Analyses include descriptive statistics and multiple regression. RESULTS: Staff members of color, regardless of position, scored higher on the thinking and behavioral styles of approval, avoidance, and competitiveness. Unlicensed assistive personnel, regardless of race or ethnicity, scored higher on the thinking and behavioral styles dependence and opposition. CONCLUSION: Nurse administrators and managers should be aware of how the addition of unlicensed assistive personnel or people of color can change the culture of the nursing unit or hospital. Explicit management strategies may be necessary to promote employee behaviors that are desirable for patient care and for the organization. PMID- 10823181 TI - Phylogenetic relation of Epidermophyton floccosum to the species of Microsporum and Trichophyton in chitin synthase 1 (CHS1) gene sequences. AB - The Nucleotide sequence of the chitin synthase 1 (CHS1) gene of Epidermophyton floccosum, an anthrophophilic dermatophyte which is the type species of the genus Epidermophyton was analyzed to determine its phylogenetic relation to eight other dermatophyte species belonging to the genera Microsporum and Trichophyton, which were sequenced in our previous studies. A genomic DNA fragment about 620 bp in length of the CHS1 gene was amplified from E. floccosum by polymerase chain reaction (PCR) and was sequenced. The CHS1 nucleotide sequence showed more than 85% similarity with sequences derived from the other dermatophytes. Phylogenetic analyses of the sequences from E. floccosum revealed that the genus Epidermophyton may be genetically distinct from Microsporum and Trichophyton. PMID- 10823182 TI - Yeasts and fluconazole susceptibility in the Philippines. AB - Identification and fluconazole susceptibility of 579 yeasts isolated from patients in 16 medical centers throughout the Philippines in 1997-98 is reported. Speciation revealed the following distribution of yeasts (with percent occurrence): Candida albicans (49.6%); C. parapsilosis (17.2%); C. tropicalis (14.9%); C. glabrata (6.2%); C. pelliculosa (4.3%); C. krusei (2.4%); C. guilliermondii (1.9%); Trichosporon cutaneum (1.7%); Cryptococcus neoformans (0.9%); Candida famata (0.5%); C. lusitaniae (0.2%) and Saccharomyces cerevisiae (0.2%). Using an agar disk diffusion assay for fluconazole susceptibility it was determined that 94% of the isolates were susceptible, 5% dose-dependent susceptible and only 1% resistant. All isolates of C. albicans were susceptible (one being dose dependent sensitive) to fluconazole. The only yeasts resistant to fluconazole were: C. guillermondii (1 isolate), C. glabrata (3 isolates) and C. parapsilosis (1 isolate). PMID- 10823183 TI - Antibiotic secondary metabolites of Dichotomomyces cejpii. AB - Nine fungal strains of Dichotomomyces cejpii were studied for their ability to produce antibiotic metabolites. It was found that they produced secondary exo- and endometabolites with antimicrobial activity against Gram-negative and Gram positive bacteria, yeasts and moulds and with a toxic effect against animal organs in vitro. Detailed chemical characterization of these active principles need to be carried out. PMID- 10823184 TI - Molecular confirmation of a Trichophyton violaceum isolate from human black-dot ringworm. AB - A clinical isolate from a black-dot ringworm lesion of a 28-year-old female Japanese was investigated by morphological and biochemical analyses as well as molecular analyses. The isolate grew well on thiamine enriched agar and did not produce violet pigment, macro-conidia or micro-conidia on Sabouraud's dextrose agar. Approximately 620-bp genomic DNA fragments of the CHS1 gene were amplified from Trichophyton mentagrophytes, T. rubrum, T. tonsurans and T. violaceum by polymerase chain reaction (PCR) and sequenced. The chitin synthase 1 (CHS1) nucleotide sequences of the clinical isolate showed more than 97% similarity to that of T. violaceum and less than 96% similarity to that of T. mentagrophytes, T. rubrum and T. tonsurans. The phylogenetic analysis of their sequences revealed that the clinical isolate was genetically close to T. violaceum and distinct from T. mentagrophytes, T. rubrum and T. tonsurans. Therefore, the isolate was confirmed as T. violaceum by mycological examination and molecular analyses. PMID- 10823186 TI - Evaluation of the mycological status of luncheon meat with special reference to aflatoxigenic moulds and aflatoxin residues. AB - The luncheon meat samples analyzed, which were produced locally by the two main luncheon meat producing companies in Egypt were relatively highly contaminated either by moulds and yeasts in general, aflatoxigenic species and aflatoxin residues in particular. The most frequently encountered fungi from the samples were yeasts, Aspergillus niger, A. flavus, Penicillium chrysogenum, Rhizopus stolonifer, Mucor circinelloides. Less common were Cladosporium sphaerospermum, Alternaria alternata, Mycosphaerella tassiana, P. aurantiogriseum and P. oxalicum. The most important aflatoxigenic species, A. flavus, was isolated frequently. It was 10% of the total fungal isolates from both samples of the two companies. Seven luncheon meat samples out of 50 analyzed were positive for aflatoxin B1 or B1 and G1, while all samples were negative for aflatoxins B2, G2, M1 and M2. Aflatoxin B1 was detected only in 4 and 3 samples out of 25 analyzed from each of company A and B, respectively. The highest detectable level, 11.1 ppb, was recorded in a sample from company B and the least, 0.5 ppb, in a sample from company A. Aflatoxin G1, at concentration of 3.2 ppb, was detected in only one sample of the aflatoxin B1--contaminated 3 samples of company B: this sample also had the highest level of aflatoxin B1. Some luncheon meat samples had higher numbers of aflatoxigenic A. flavus than others, however these samples were negative for aflatoxins. The hazardous potential of such contamination will be discussed. PMID- 10823185 TI - Comparative efficacy of fluconazole and amphotericin B in the parenteral treatment of experimental paracoccidioidomycosis in the rat. AB - Patients with severe and complicated paracoccidioidomycosis are treated with amphotericin B by the intravenous route. Fluconazole is active in vitro against Paracoccidioides brasiliensis and can also be administered intravenously, but few clinical or experimental data are available about its action against the infection caused by this fungus. In the present study, the efficacy of fluconazole and amphotericin B was assessed comparatively in rats inoculated parenterally with P. brasiliensis. The treatment was performed 3 times a week for 4 weeks starting one week after infection. Fluconazole administered intraperitoneally (14 mg/kg body weight/dose) was more effective (P < 0.001) than amphotericin B (2 mg/kg body weight/dose) in reducing the number of colony forming units in the lungs and spleen. When administered intravenously at the dose of 3 mg/kg body weight, fluconazole was as effective as amphotericin B (0.8 mg/kg body weight) in reducing the pulmonary fungal burden. Under these conditions, the rats treated with fluconazole had a smaller number of colony forming units than untreated animals (P < 0.001), but amphotericin B was more effective than fluconazole in reducing spleen infection (P < 0.005). Except for this result obtained with a low dose, fluconazole showed an antifungal action equal to or higher than that of amphotericin B. The activity of fluconazole at doses equivalent to those used for human treatment suggests that this antifungal agent may be an alternative to amphotericin B for the early intravenous treatment of patients with paracoccidioidomycosis. PMID- 10823187 TI - An unprecedented nitrogen elimination reaction: mechanistic studies using 15N labeled 4-amino-7-benzylpyrrolo[2,3-d][1,2,3]triazine-5-carbonitrile. AB - [formula: see text] In the course of our work on novel pyrrolo[2,3 d][1,2,3]triazines we have discovered that 1 undergoes an elimination of nitrogen at 250 degrees C to give 2. We have conducted 15N labeling studies that establish that the loss of N-2 and N-3 from 1 had occurred rather than N-1 and N-2, presumably via a retro Diels-Alder reaction of the imino tautomer 7. This study provides an alternative to the commonly accepted mechanism which involves the loss of N-1 and N-2 via the transient formation of a diazonium compound generated from 4-amino- or 4-oxo-substituted 1,2,3-triazines condensed with carbocycles or heterocycles. PMID- 10823188 TI - Catalytic enantioselective amination of enolsilanes using C2-symmetric copper(II) complexes as chiral Lewis acids. AB - [formula: see text] [Cu(S,S)-t-Bu-box](OTf)2 (1) catalyzes the enantioselective amination of enolsilanes with azodicarboxylate derivatives. Isomerically pure enolsilanes of aryl ketones, acylpyrroles, and thioesters added to the azo-imide in greater than 95% ee. The use of an alcohol additive was critical to achieving catalyst turnover. PMID- 10823189 TI - Asymmetric synthesis of the balanol heterocycle via a palladium-mediated epimerization and olefin metathesis. AB - [formula: see text] The enantioselective formal synthesis of balanol, a potent protein kinase C inhibitor, was accomplished from D-serine utilizing a Pd catalyzed equilibration of diastereomeric 5-vinyloxazolines to set the stereochemistry of the vicinal amino and hydroxyl groups. A ruthenium-catalyzed ring-closing metathesis was employed to form the seven-membered nitrogen heterocyle. PMID- 10823190 TI - Stereospecific synthesis and biological evaluation of farnesyl diphosphate isomers. AB - [formula: see text] A unified, stereospecific synthetic route to the three geometric isomers of (E,E)-farnesyl diphosphate (E,E-FPP) (1, 2, and 3) has been developed. The key feature of this synthesis is the ability to control the stereochemistry of triflation of the beta-ketoester 10 to give either 11 or 14. Preliminary evaluation of these compounds with protein-farnesyl transferase indicates that 1 and 2 are surprisingly effective substrates; however, Z,Z-FPP (3) is a poor substrate and a sub-micromolar inhibitor. PMID- 10823191 TI - 5-Amino-dU: a nucleoside analogue in the central strand of DNA triplex for orientation selective binding of A/G/C/T in the third strand. AB - [formula: see text] 5-Amino-dU, a designed nucleoside analogue, when placed in the central strand of DNA triple helix, recognizes all four bases A, G, C, and T in the third strand, with a selectivity based on the orientation (parallel/antiparallel) of the third strand. PMID- 10823192 TI - A new method for the generation and cyclization of iminyl radicals via the Hudson reaction. AB - [formula: see text] A mild new synthetic procedure has been developed for in situ generation and cyclization of iminyl radicals onto pendant alkenes, followed by functionalization of the resulting carbon radical by one of a variety of trapping reagents. The key process in the method involves production of the iminyl radical via treatment of an aldoxime or ketoxime with readily available 2,6 dimethylbenzenesulfinyl chloride at -50 degrees C to room temperature (Hudson reaction). PMID- 10823193 TI - Hetero [4 + 2] cycloadditions of (trialkylsilyl)vinylketenes. Synthesis of alpha,beta-unsaturated delta-valerolactones and -lactams. AB - [formula: see text] Hetero Diels-Alder reactions of (trialkylsilyl)vinylketenes (TAS-vinylketenes) with carbonyl and imino dienophiles are described. TAS vinylketenes participate as electron-rich dienes in [4 + 2] cycloadditions with diethyl ketomalonate to afford alpha,beta-unsaturated delta-valerolactones in good yield. Nonenolizable N-alkyl- and N-(trimethylsilyl)imines combine with TAS vinylketenes to furnish alpha,beta-unsaturated delta-valerolactams. In contrast to most Diels-Alder reactions involving unactivated imines, these cycloadditions do not require promotion by Lewis acids and in general proceed with a high degree of stereoselectivity. PMID- 10823194 TI - Postulated biogenesis of WS9885B and progress toward an enantioselective synthesis. AB - [formula: see text] WS9885B promotes the assembly of microtubules in vitro and displays cytotoxicity as potent as paclitaxel against several cancer cell lines. In this Letter, we propose a biogenesis for this architecturally complex bacterial metabolite from a much simpler, polyunsaturated precursor. We also present significant progress toward a convergent, enantioselective synthesis of WS9885B. Our work features a chemoselective palladium-catalyzed cross-coupling of two advanced building blocks and an uncommon Claisen-like cyclization. PMID- 10823195 TI - A formal total synthesis of roseophilin: cyclopentannelation approach to the macrocyclic core. AB - [formula: see text] The formal total synthesis of the macrocyclic core of roseophilin 4 has been accomplished in 12 steps from 5-hexenal 8. The cyclopentannelation reaction was used to form the aliphatic five-membered ring of 3. Diene 2 was assembled by means of a Stetter reaction. Ring-closing metathesis of 2, reduction, and Knorr reaction of the 1,4-diketone 5 gave the ketopyrrole 3. PMID- 10823197 TI - Enantiopure N-acyldihydropyridones as synthetic intermediates: asymmetric synthesis of benzomorphans. AB - [formula: see text] Concise asymmetric syntheses of several benzomorphan derivatives have been accomplished using enantiopure 2,3-dihydro-4-pyridones as chiral building blocks. PMID- 10823196 TI - Unusual conformational effect in alpha-aminoorganostannanes. AB - [formula: see text] Dynamic NMR analysis of conformationally mobile and rigid 2 tributylstannyl-N-methylpiperidines revealed an unexpected conformational effect that is manifested in a small energy difference between conformers in which the tin is equatorial and axial. The major reason appears to be a distortion of the conformer in which the C-2-Sn bond is synclinal to the nitrogen lone pair. PMID- 10823198 TI - Total synthesis of (+/-)-biphyscion. AB - [formula: see text] A regiospecific total synthesis of (+/-)-biphyscion (1) is described. A novel aspect of the plan was construction of the bianthraquinone ring system from a biphenyl intermediate through the use of a one-pot, double isobenzofuranone condensation. PMID- 10823199 TI - Regiocontrolled synthesis of carbocycle-fused indoles via arylation of silyl enol ethers with o-nitrophenylphenyliodonium fluoride. AB - [formula: see text] A new, regiocontrolled synthesis of carbocycle-fused indoles has been developed. The two-step procedure involves first the regiospecific arylation of silyl enol ethers with o-nitrophenylphenyliodonium fluoride (1). Reduction of the nitro group on the aromatic ring with TiCl3 followed by spontaneous condensation of the aniline with the ketone then affords the indole products. PMID- 10823200 TI - Selective phosphitylation of the primary hydroxyl group in unprotected carbohydrates and nucleosides. AB - [reaction: see text] Carbohydrates and nucleosides containing a phosphate at the less-hindered primary hydroxyl group are often prepared using a protection/deprotection strategy. Herein we report that the phosphoramidite method can be used to selectively incorporate phosphorus at the primary hydroxyl group of O-unprotected carbohydrates and nucleosides; in situ oxidation of the resulting phosphite triester yields the phosphate triester. PMID- 10823201 TI - Intramolecular acid-catalyzed amide isomerization in aqueous solution. AB - [reaction: see text] We report for the first time that stoichiometric and even catalytic quantities of weak acids in aqueous solution can very efficiently catalyze amide isomerization in a carefully designed system in which a proton donor is situated so that intramolecular hydrogen bonding to the amide nitrogen is highly favored. Our results provide the first experimental verification that hydrogen bond donation to the amide nitrogen by charged proton donors may play a very significant role in the enzymatic catalysis of amide isomerization. PMID- 10823202 TI - Creation of a synthetically useful lipase with higher than wild-type enantioselectivity and maintained catalytic activity. AB - [reaction: see text] We have found that two Geotrichum candidum lipase isozymes have remarkably different abilities to differentiate between enantiomers of ethyl 2-methyldecanoate. By rational recombination of selected portions of the two isozymes, we have created a novel lipase with an enantioselectivity superior to that of the best wild-type parent isozyme. Site-directed mutagenesis identified two key amino acid residues responsible for the improved enantioselectivity without compromised total activity of the reengineered enzyme. PMID- 10823203 TI - Anomer-selective inhibition of glycosidases using aminocyclopentanols. AB - [reaction: see text] (1S,2S,3S,4R,5R)-4-amino-5-(hydroxymethyl)cyclopentane-1,2,3 triol 1 is prepared stereoselectively from D-lyxose and displays anomer-selective inhibition for beta-galactosidase (Ki = 3.0 x 10(-6) M) and beta-glucosidase (Ki = 1.5 x 10(-7) M), over alpha-galactosidase (Ki = 2.3 x 10(-5) M) and alpha glucosidase (IC50 = 1.0 x 10(-4) M). There is no observable cross-reactivity with alpha-mannosidase, beta-mannosidase, or alpha-L-fucosidase. PMID- 10823204 TI - Synthesis of a chemiluminescent acridinium hydroxylamine (AHA) for the direct detection of abasic sites in DNA. AB - [structure: see text] The synthesis of a chemiluminescent acridinium hydroxylamine (AHA) for the direct detection of abasic sites in damaged nucleic acids is described. The reagent reacts readily with abasic sites of damaged calf thymus DNA generated in a time-dependent manner under acid/heat depurination conditions. Preliminary results indicate the sensitivity of the direct chemiluminescent detection format is approximately 0.1 abasic sites detected per 10(6) nucleotides using as little as 200 ng of DNA. PMID- 10823205 TI - Tert-butylsulfonamide. A new nitrogen source for catalytic aminohydroxylation and aziridination of olefins. AB - [reaction: see text] The N-chloramine salt of tert-butylsulfonamide has been shown to be an efficient nitrogen source and the terminal oxidant for catalytic aminohydroxylation and aziridination of olefins, resembling closely Chloramine-T by its behavior in these catalytic reactions. The sulfonyl-nitrogen bond in the product or its derivatives is easily cleaved under mild acidic conditions, allowing for facile liberation of the amino group. PMID- 10823206 TI - Synthesis of a protected 3,4-dihydroxyproline from a pentose sugar. AB - [reaction: see text] D-Ribonolactone (6) was transformed into N ((fluorenylmethoxy)carbonyl)-3,4-bis-O-(tert-butyldimethylsilyl)-D-2,3-cis-3,4 cis-3,4-dihydroxyproline (13) in nine chemical steps. This represents a potentially general strategy for the synthesis of 3,4-dihydroxyprolines, utilizing the pentose sugar series as starting materials. PMID- 10823207 TI - Beta-lactones as a new class of cysteine proteinase inhibitors: inhibition of hepatitis A virus 3C proteinase by N-Cbz-serine beta-lactone. AB - [reaction: see text] N-Benzyloxycarbonyl-L-serine beta-lactone (1) is shown to irreversibly inactivate the 3C cysteine proteinase of hepatitis A virus (HAV) with k(inact) = 0.70 min(-1), K(I) = 1.84 x 10(-4) M and k(inact)/K(I) = 3800 M( 1) min(-1) at an enzyme concentration of 0.1 microM. Mass spectrometric and HMQC NMR studies using 13C-labeled 1 show that the active site cysteine (Cys-172) thiol of the HAV 3C proteinase attacks the beta-position (i.e. C-4) of the oxetanone ring, thereby leading to ring opening and alkylation of the sulfur. In contrast, the enantiomer of this beta-lactone, 2, is a reversible competitive inhibitor (Ki = 1.50 x 10(-6) M) at similar enzyme concentrations. The beta lactone motif represents a new class of inhibitors of cysteine proteinases. PMID- 10823208 TI - A facile method for the solution and solid-phase synthesis of substituted [3.3.1] bicycles. AB - [reaction: see text] Interest in bicyclic natural products from the Guttiferae classification has led to the development of an improved method for the selenium mediated cyclization of alkenyl-substituted beta-dicarbonyls (I) to form a variety of bicyclo[3.3.1]nonan-9-ones (II) both in solution and on solid support. PMID- 10823209 TI - Simple and practical routes to enantiomerically pure 5-(trialkylsilyl)-2 cyclohexenones. AB - [reaction: see text] Enantiomerically pure chiral 5-silylated 2-cyclohexenones are easily prepared in high yield using as a key step kinetic resolution with a commercially available lipase. Fully active enzyme can be recovered very efficiently for reuse. The synthetic steps are outlined in Schemes 1 and 3. Enantiomerically pure 2-cyclohexenones such as 1 and 2 are versatile intermediates for the synthesis of a multitude of chiral targets by means of a variety of diastereoselective reactions such as those illustrated in Scheme 2. PMID- 10823211 TI - A simple convergent approach to the synthesis of the antiviral agent virantmycin. AB - [structure: see text] The antiviral agent (+/-)-virantmycin has been synthesized from two simple building blocks (2 and 3) in eight steps, as outlined in Scheme 2. PMID- 10823210 TI - Yeast protein farnesyltransferase. Binding of S-alkyl peptides and related analogues. AB - [reaction: see text] Protein farnesyltransferase (PFTase) catalyzes alkylation of cysteine residues by farnesyl diphosphate (FPP). The dissociation constants for the PFTase-peptide analogue complexes for the series of analogues fl-RTRC(X)VIA (X = H, methyl, dodecyl, farnesyl) were measured by fluorescence anisotropy. The results indicate that an ionizable sulfhydryl moiety is important for substrate binding and the farnesyl group in the product facilitates binding. PMID- 10823212 TI - Catalysis and acceleration of acyl transfer by aminocyclodextrins: a biomimetic system of use in enzyme modeling and drug design. AB - [formula: see text] Aminocyclodextrins (ACDs), perfunctionalized at the 6 position with amino groups, bind phosphonoformate (PFA) diesters and accelerate acyl transfer reactions with high efficiency at neutral pH. Aminolysis and esterolysis are accelerated and hydrolysis of PFA diesters is catalyzed by ACDs. PFA diesters have significant antiviral activity. The rapid reactions observed with ACDs show that biological nucleophiles may undergo facile covalent modification by PFA esters at physiological pH, which has significant implications for prodrug and drug design strategies. PMID- 10823213 TI - A chiral rhodium complex for rapid asymmetric transfer hydrogenation of imines with high enantioselectivity. AB - [formula: see text] A chiral rhodium complex, (R)-Cp*RhCl[(1S,2S)-p TsNCH(C6H5)CH(C6H5)NH2] (1a, (S,S)-Cp*RhClTsDPEN), generated from [Cp*RhCl2]2 and (1S,2S)-N-p-toluenesulfonyl-1,2-diphenylethylenediamine [(S,S)-TsDPEN], and its enantiomer 1b were found to provide superior catalysts for the rapid, high yielding, asymmetric transfer hydrogenation of some heterocyclic imines, using an HCO2H-Et3N azeotrope as the hydrogen source. PMID- 10823214 TI - Absolute configurations of N,N-dialkyl alpha-amino acids and beta-amino alcohols from exciton-coupled circular dichroism spectra of Cu(II) complexes. AB - [formula: see text] A circular dichroism technique has been applied to the absolute configurational assignment of acyclic alpha-amino acids and beta-amino alcohols with single stereogenic centers via a one-step derivitization procedure requiring microgram quantities of material. Metal ions fix the geometrical relationship of two chromophores, affording CD spectra that agree with theory on the basis of the expected conformation of the ligands in the metal complexes. PMID- 10823215 TI - Catalytic enantioselective Michael additions to unsaturated ester derivatives using chiral copper(II) Lewis acid complexes. AB - [formula: see text] Chiral Cu(II) bisoxazoline (box) Lewis acids have been developed as catalysts of the Michael addition of enolsilanes to unsaturated ester derivatives. While enantioselection is stereoregular, the sense of diastereoselection is directly related to thioester enolsilane geometry: (E) enolsilanes give anti adducts and (Z) enolsilanes afford syn adducts. The size of the enolsilane alkylthio substituent directly impacts the magnitude of diastereoselection. PMID- 10823216 TI - Cytotoxic polyprenylated benzoylphloroglucinol derivatives with an unusual adamantyl skeleton from Hypericum sampsonii (Guttiferae). AB - [formula: see text] The structures of sampsoniones I and J, isolated from the aerial parts of the Chinese medicinal plant Hypericum sampsonii, have been elucidated by detailed spectral analysis. They are complex adamantyl derivatives, and sampsonione I is the first polyprenylated benzoylphloroglucinol derivative with the unique caged tetracyclo[7.3.1.1.0]tetradecane-2,12,14-trione skeleton. Cytotoxic sampsonione I has also been obtained by the biomimetic transformation of sampsonione J. PMID- 10823217 TI - New synthetic technology for the synthesis of hindered alpha-diazoketones via acyl mesylates. AB - [formula: see text] A mild and reliable one-pot protocol for the elaboration of sterically demanding carboxylic acids into alpha-diazoketones via acyl mesylates has been developed. Aside from delineating the reaction parameters which render this strategy quite general for hindered carboxylic acids, we have directly proven the existence of the fleeting acyl mesylate group as the reactive species in these reactions and shed light onto the differing mechanisms which are operative in the activation of hindered and simple carboxylic acids with methanesulfonyl chloride. PMID- 10823218 TI - 2-Deoxy-2-iodo- and 2-deoxy-2-bromo-alpha-glucopyranosyl trichloroacetimidates: highly reactive and stereoselective donors for the synthesis of 2-deoxy-beta glycosides. AB - [formula: see text] 2-Deoxy-2-iodo- and 2-deoxy-2-bromoglucopyranosyl trichloroacetimidates 8-10 and 22 are extremely useful precursors of 2-deoxy-beta glycosides. These reactive glycosyl donors undergo highly stereoselective glycosidation reactions at -78 degrees C with a range of glycosyl acceptors using TBS-OTf as the activating agent. beta-Glycosides are obtained with > or = 19:1 selectivity in six of the seven examples reported herein. PMID- 10823219 TI - Iodoacetoxylation of glycals using cerium(IV) ammonium nitrate, sodium iodide, and acetic acid: stereoselective synthesis of 2-deoxy-2-iodo-alpha-mannopyranosyl acetates. AB - [formula: see text] The reactions of glycals with ceric(IV) ammonium nitrate and sodium iodide in the presence of acetic acid provides 2-deoxy-2-iodo-alpha mannopyranosyl acetates with good stereoselectivity. In the majority of the cases examined, the selectivity was considerably better than that from reactions using N-iodosuccinimide and HOAc. PMID- 10823220 TI - 2-Deoxy-2-iodo-alpha-mannopyranosyl and -talopyranosyl acetates: highly stereoselective glycosyl donors for the synthesis of 2-deoxy-alpha-glycosides. AB - [formula: see text] TMS-OTf- or TBS-OTf-promoted glycosidation reactions of 2 deoxy-2-iodo-alpha-mannopyranosyl acetates 8-10 and the 2-deoxy-2-iodo-alpha talopyranosyl acetate 11 provide the corresponding 2-deoxy-2-iodo-alpha pyranosides, precursors to 2-deoxy-alpha-glycosides, as the only observed reaction products. PMID- 10823221 TI - Phorboxazole synthetic studies. 1. Construction of a C(3-19) subtarget exploiting an extension of the Petasis-Ferrier rearrangement. AB - [formula: see text] In this, the first of two letters, we outline our overall strategy for the total synthesis of phorboxazoles A (1) and B (2), rare oxazole containing macrolides possessing extraordinary antimitotic activity, and describe the assembly of a C(3-19) subtarget (-)-5 for the total synthesis of phorboxazole A. The synthesis of (-)-5 was achieved in 15 linear steps (12% overall yield), exploiting a modification of the Petasis-Ferrier rearrangement to construct the C(11-15) cis-tetrahydropyran. Dimethylaluminum chloride (Me2AlCl) proved to be the Lewis acid of choice for the Petasis-Ferrier rearrangement. PMID- 10823222 TI - Phorboxazole synthetic studies. 2. Construction of a C(20-28) subtarget, a further extension of the Petasis-Ferrier rearrangement. AB - [formula: see text] In this, the second of two Letters, we describe the efficient assembly of (+)-4, a C(20-28) subtarget for the total synthesis of phorboxazoles A (1) and B (2). The synthesis was achieved in 12 linear steps (20% overall yield) via Petasis-Ferrier rearrangement of an E/Z mixture of trisubstituted enol ethers (15) to assemble the C(22-26) cis-tetrahydropyran. A mechanism for the observed diastereoconvergence of 15 is proposed. In addition, a new tactic for the synthesis of enol ethers (e.g., 15) based on the elegant work of Julia is described. PMID- 10823223 TI - Stereochemistry of the reduction step mediated by recombinant 1-deoxy-D-xylulose 5-phosphate isomeroreductase. AB - [formula: see text] The stereochemistry of the 1-deoxy-D-xylulose 5-phosphate (DXP) isomeroreductase reduction step has been examined using the recombinant enzyme from Synechocystis sp. PCC6803. Using [3-2H]DXP and [4S-2H]NADPH, it has been determined that the C1 pro-S hydrogen in the 2-C-methyl-D-erythritol 4 phosphate product derives from C3 of DXP, indicating that hydride attack occurs on the re face of the intermediate aldehyde. The 4S-hydride from NADPH is delivered, assigning this enzyme as a class B dehydrogenase. PMID- 10823224 TI - Enzymatic selective dehydration and skeleton rearrangement of paclitaxel precursors. AB - [formula: see text] Enzymatic dehydration reactions to remove selectively the chemically least reactive 13-hydroxyl group of 10-deacetylbaccatin III and its derivatives 2a-c are described. The elimination of the 13-OH takes place with skeleton rearrangement, allowing us to synthesize some interesting diterpenoids in good yields. PMID- 10823225 TI - Total synthesis of reveromycin B. AB - [formula: see text] The stereoselective total synthesis of reveromycin B (2), a novel polyketide-type antibiotic, has been accomplished. PMID- 10823226 TI - A general route to 4-imidazolyl-containing multidentate ligands for biomimetic studies. AB - [formula: see text] 4-iodo-1-tritylimidazole undergoes magnesium-iodine exchange with a Grignard reagent to give selectively the 4-magnesioimidazole derivative, which reacts with esters to form a variety of poly-4-imidazolyl carbinol compounds in 40-79% yields. A wide range of bi-, tri-, and pentadentate ligands featuring 4-substituted imidazole units have been efficiently synthesized. PMID- 10823227 TI - Synthesis and activity of a new generation of ruthenium-based olefin metathesis catalysts coordinated with 1,3-dimesityl-4,5-dihydroimidazol-2-ylidene ligands. AB - [formula: see text] A new family of 1,3-dimesityl-4,5-dihydroimidazol-2-ylidene substituted ruthenium-based complexes 9a-c has been prepared starting from RuCl2(=CHPh)(PCy3)2 2. These air- and water-tolerant complexes were shown to exhibit an increased ring-closing metathesis activity at elevated temperature when compared to that of the parent complex 2 and the previously developed complex 3. In many instances the activity of these complexes also rivaled or exceeded that of the alkoxy-imido molybdenum complex 1. Catalyst loadings of as low as 0.05 mol% could be used. PMID- 10823228 TI - Risk factors for gonococcal and chlamydial cervical infection in pregnant and non pregnant women in Zimbabwe. AB - OBJECTIVE: To identify demographic, behavioural and clinical characteristics of symptomatic and asymptomatic women with gonococcal and/or chlamydial cervicitis a study was conducted among women attending antenatal clinics and primary care clinics in Harare, Zimbabwe. DESIGN: Cross sectional study. SETTING: Primary care clinics and antenatal clinics in Harare. SUBJECTS: 467 women with vaginal discharge and 1,189 asymptomatic pregnant women. MAIN OUTCOME MEASURES: Behavioural and clinical correlates of gonococcal and chlamydial cervical infection. RESULTS: The mean age of symptomatic women was 26.11 +/- 6.84 years (range: 15 to 52 years) and that of asymptomatic pregnant women was 24.67 +/- 5.43 years (range: 15 to 45 years). Gonococcal and/or chlamydial cervical infection was found in 69 of 1,189 (5.8%) pregnant women and in 77 of 467 (16.5%) non-pregnant women. Logistic regression analysis identified the following predictors of gonococcal or chlamydial infection in women with vaginal discharge: being separated from the partner for a month or more (p = 0.002), having had sex with a new partner in the last three months (p = 0.002), current use of condoms (p = 0.011), and the finding on examination of a purulent vaginal discharge (p = 0.004). Amongst these women an increasing educational level was inversely associated with cervical infection (p = 0.007). Amongst asymptomatic pregnant women the following were identified as predictors of cervical infection: the patient admitting to having a vaginal discharge on direct questioning (p = 0.004), and the finding of a purulent vaginal discharge on examination (p = 0.001). CONCLUSIONS: Amongst symptomatic and asymptomatic women certain behavioural factors and some clinical findings are associated with cervical gonococcal or chlamydial infection. Women with multiple partners and with partners who are currently using condoms with them and those women with a purulent vaginal discharge are likely to be infected. The age and marital status of subjects was not associated with cervical infection. These findings are useful in providing appropriate care for women with overt or minimal symptoms. PMID- 10823229 TI - Pattern of admissions to the paediatric medical wards (1995 to 1996) at Harare Hospital, Zimbabwe. AB - OBJECTIVE: To document the pattern of disease and outcome of medical paediatric admissions at Harare hospital. DESIGN: Cross sectional study from 1 June 1995 to 30th May 1996. SETTING: Medical paediatric wards, Harare Central Hospital. SUBJECTS: All admissions below the age of nine years. MAIN OUTCOME MEASURES: Socio-demographic features, nutritional status, clinical diagnoses, duration of hospital stay and mortality. RESULTS: A total of 8,826(90.0%) of the admissions were assessed. The majority(51.8%) of the patients were one year and below. "Road to Health" cards, available for 94.4% children, showed 88.0% had appropriate immunizations for age. Forty nine percent had lower respiratory tract infection either as only, or concurrent with other diagnoses, with an overall fatality rate of 16.2%. Acute diarrhoea was the second most frequent condition (21.0%) with an associated case fatality rate of 11.3%. Mortality rates for neonatal sepsis and bacterial meningitis were 12.3% and 32.8% respectively. Severe malnutrition defined as less than 60% of the expected weight for age, was present in 12.5% with an associated fatality rate of 28.7%. Based on clinical suspicion 23.2% of the total children were tested for HIV antibodies and 82.0% were positive. Overall case fatality among the inpatients was 17.8% and 36.7% of the total deaths occurred within 24 hours of admission. CONCLUSION: There is a need to re evaluate our management strategies for common conditions including those with HIV infection. Standardization of case management is an important and essential step towards reduction in mortality both at the primary and referral health institutions. PMID- 10823230 TI - Plantar and digital dermatoglyphics in Malawi. AB - OBJECTIVE: To establish plantar and digital dermatoglyphic patterns. DESIGN: Cross sectional study of randomly selected Black Malawian subjects. SETTING: Able bodied indigenous Black subjects were recruited from secondary schools and the College of Medicine in Blantyre city. MAIN OUTCOME MEASURES: Dankmeijer's index (DI) and the pattern intensity (PII) indices were determined and the variability of ridge patterns were counted and classified. MATERIALS AND METHODS: Bilateral plantar and digital prints of the sole of selected subjects were recorded, studied and classified. RESULTS: Arches were the most predominant ridge pattern found on the digits. Whorls were absent on all digits, a feature which appears to be peculiar to Malawians. Loops were also absent on the small digit but present on the big toe. These were significantly greater in males than females (p < 0.05). The PII was higher in males than females while, DI was higher in females than in males. These indices were significantly different from similar values obtained from previous studies of two ethnic groups in Nigeria (p < 0.05). CONCLUSION: This study has provided the normal dermatoglyphic patterns of Malawian subjects and also highlighted that digital patterns are more specific in differentiating tribes and population groups. PMID- 10823231 TI - Hypotensive effect of crude extract Olea. africana (Oleaceae) in normo and hypertensive rats. AB - OBJECTIVE: To investigate the effects of crude extracts of the root and stem of Olea africana on mean arterial pressure (MAP) and heart rate in normo and hypertensive rats. DESIGN: Comparative study. SETTING: Laboratory. SUBJECTS: 40 Sprague Dawley rats of either sex (150 to 250 g) randomly assigned to normotensive (n = 15) (DOCA) -salt hypertensive (n = 25) groups. The normotensive group comprised three subgroups (n = 5) namely, a control treated with graded volumes of normal saline intravenously (i.v.), and two groups respectively treated with graded doses of aqueous extract of O. africana (i.v.). The hypertensive group comprised five subgroups (n = 5) namely, a control group of sham operated rats, and four groups of nephrectomised rats. One group of nephrectomized rats received Deoxycorticosterone acetate (DOCA) and salt for seven days, while others received DOCAsalt for 14 days. Two groups of the 14 day treated rats respectively received 200 mg/kg and 1,000 mg/kg of O. africana orally thereafter. MAP and HR were measured under sodium pentobarbital anaesthesia (35 mg/kg intraperitoneally). MAIN OUTCOME MEASURES: Mean arterial pressure and heart rate. RESULTS: Intravenous administration of aqueous and ethanolic extracts of O. africana caused an immediate and dose dependent fall in MAP and HR in anaesthetised normotensive rats. The aqueous extract was more potent than the ethanolic extract. Orally administered aqueous extract produced lowering of MAP and HR in DOCA-salt hypertensive rats. Propranolol partially blocked the Map lowering effect of O. africana. CONCLUSION: Extracts of O. africana lowered MAP and HR in normotensive and DOCA-salt hypertensive rats. Hence the use of the aqueous extracts in the treatment of some forms of hypertension and heart palpitations in humans. This effect may be mediated via beta adrenergic receptors. PMID- 10823232 TI - Tachycardiomyopathy--a case report. AB - This case report describes a patient presenting with atrial tachyarrhythmia and severe left ventricular failure in which rate control was associated with restoration of normal contractile function. PMID- 10823233 TI - Management of migraine--no longer a headache. AB - There have been important advances in the management of migraine headache. Revised diagnostic criteria, which were introduced in 1988 1 have led to more reliable assessment of the prevalence of migraine and the way it affects attendance and efficiency at work. In addition, the use of the 5HTI agonists has become established. The correct diagnosis of migraine and other headaches is of greatest importance as unless the right diagnosis is made, the right treatment cannot be given. In this article, we discuss how these changes have altered the management of migraine in adults. PMID- 10823234 TI - Freewheeling centrioles. AB - The century-old controversy over the reproduction and function of the centriole is examined to elucidate the conceptual and methodological issues that have made it so resistant to closure. The study of centrioles is situated in two distinct eras punctuated by the deployment of the electron microscope. From the late 19th century to the mid-20th century, centrioles were defined largely in functional terms- as self-reproducing 'central bodies' playing a directive role in mitosis. During this period, when their structure remained unknown, their universal presence in all cells was ambiguous, and their reality was seriously debated. A conceptual switch occurred after the mid-1950s. When the centriole was made visible under the electron microscope, it was defined in terms of its characteristic cart-wheel structure, but its function and manner of reproduction have remained enigmatic. The controversy over the nature of centrioles illustrates the dynamic interplay of techniques, theories, and background assumptions in the production of scientific knowledge. It also highlights the difficulties biologists face in coming to grips with problems of cell structure and intracellular morphogenesis. PMID- 10823235 TI - Constant hybrids in Mendel's research. AB - The persisting controversial interpretation of constant hybrids and of the term Entwicklungsgeschichte, mentioned by Mendel in the Pisum paper, is elucidated in the context of his experiments with other plant species and of the growth of knowledge in scientific animal and plant breeding in Moravia. PMID- 10823236 TI - From cell lineage to developmental genetics. AB - One of the bases of developmental genetics resides in the alliance of clonal analysis and genetic analysis. But the study of cell lineage--cells which have their genealogical relationship--and the study of the cellular labelled progeny, have their own history. We have tried to follow it since its foundation with C.O. Whitman (1878) and E.B. Wilson (1892). A.H. Sturtevant (1929) and C. Stern (1936) the first tools to study the 'cell lineage' in Drosophila. We stress the contribution of the pioneer work realised around 1940. In the following period we witness the emergence of developmental genetics in Drosophila mainly with E. Hadorn (1949-1966), C. Stern (1954-1968) and E.B. Lewis (1963-1964). We conclude with A. Garcia-Bellido's view of compartments: supra-cellular units of development (1973). A postscript presents the most recent publications and some critical focuses. PMID- 10823237 TI - Calcium signalling: a historical account, recent developments and future perspectives. AB - Ca2+ is a uniquely important messenger that penetrates into cells through gated channels to transmit signals to a large number of enzymes. The evolutionary choice of Ca2+ was dictated by its unusual chemical properties, which permit its reversible complexation by specific proteins in the presence of much larger amounts of other potentially competing cations. The decoding of the Ca2+ signal consists in two conformational changes of the complexing proteins, of which calmodulin is the most important. The first occurs when Ca2+ is bound, the second (a collapse of the elongated protein) when interaction with the targeted enzymes occurs. Soluble proteins such as calmodulin contribute to the buffering of cell Ca2+, but membrane intrinsic transporting proteins are more important. Ca2+ is transported across the plasma membrane (channel, a pump, a Na+/Ca2+ exchanger) and across the membrane of the organelles. The endoplasmic reticulum is the most dynamic store: it accumulates Ca2+ by a pump, and releases it via channels gated by either inositol 1,4,5-trisphosphate (IP3) and cyclic adenosine diphosphate ribose (cADPr). The mitochondrion is more sluggish, but it is closed-connected with the reticulum, and senses microdomains of high Ca2+ close to IP3 or cADPr release channels. The regulation of Ca2+ in the nucleus, where important Ca(2+) sensitive processes reside, is a debated issue. Finally, if the control of cellular Ca2+ homeostasis somehow fails (excess penetration), mitochondria 'buy time' by precipitating inside Ca2+ and phosphate. If injury persists, Ca2(+) death eventually ensues. PMID- 10823238 TI - Nuclear calcium signalling. AB - The topic of nuclear Ca2+ signalling is beset by discrepant observations of substantial nuclear/cytoplasmic gradients. The reasons why some labs have recorded such gradients, whilst other workers see equilibration of Ca2+(cyt) and Ca2+(nuc) using the same cells and techniques, is unexplained. Furthermore, how such gradients could arise across the NE that possesses many highly-conductive NPCs is a mystery. Although nuclei may have the capacity to be autonomous signalling entities, with functional Ca2+ release channels and an inositide cycle, the balance of evidence suggests that Ca2+ release on the inner NE does not occur during physiological stimulation. Our work suggests that elementary Ca2+ release events originating in the cytoplasm can give rise to Ca2+ signals without causing elevation of the bulk cytoplasm. Clearly, the many Ca2+ signalling mechanisms that may impinge on Ca2+(nuc) will remain a topic of controversy and debate for some time. PMID- 10823239 TI - Nuclear inositides: inconsistent consistencies. AB - It is now clear that phosphoinositides, which play a major role in the regulation of a variety of cellular processes in the cytoplasm, are found within the nucleus. Their role in this subcellular compartment is still contentious: however, data has suggested that nuclear inositides generate substrates, such as PtdIns(4,5)P2, utilised by a number of nuclear signalling pathways: for example, nuclear phospholipase C and the PtdIns 3-kinase cascade. There is also evidence that PtdIns(4,5)P2 may play a role in the localisation and regulation of a number of nuclear proteins such as the BAF complex, which is involved in the regulation of chromatin structure. Although the presence of nuclear inositides has been demonstrated in a number of different cell types, suggesting that it is ubiquitous, there are many inconsistencies within the literature concerning the locations and isotypes of enzymes that are involved in their regulation and in the potential second messengers which are generated by them. This review aims to highlight some of these inconsistencies in order to focus on areas that need further characterisation. PMID- 10823240 TI - Calcium regulation of the brain-derived neurotrophic factor gene. AB - Results from several laboratories have suggested that peptide factors known as neurotrophins may play roles coupling changes in synaptic activity to lasting changes in synaptic function. Consistent with this idea, increases in synaptic activity and intracellular calcium induce the expression of the gene that encodes the neurotrophin, brain-derived neurotrophic factor. Recently, a pathway has been elucidated in neurons by which the influx of extracellular calcium evokes brain derived neurotrophic factor transcription (BDNF). Calcium activates BDNF transcription through two adjacent calcium response elements within one of the promoters of the BDNF gene. One of the two elements binds to the cyclic adenosine monophosphate (AMP) response element binding protein (CREB) transcription factor, and interfering with CREB or related family members inhibits calcium-dependent BDNF transcription. This review focuses on the mechanisms by which calcium influx regulates brain-derived neurotrophic factor expression and the implications that these results have for potential roles of neurotrophins in synaptic function. PMID- 10823241 TI - Calcium-regulated protein kinase cascades and their transcription factor targets. AB - In the nervous system, calcium signals associated with electrical activation of neurons induce gene transcription that may be important for long-lasting adaptation. The type of transcriptional response is determined by the properties of the calcium signal that include subcellular localisation, amplitude, duration and the physical site of entry. Here we review calcium-regulated protein kinase cascades and discuss potential mechanisms through which they propagate calcium signals to and within the nucleus and control the activity of transcription factors and transcriptional co-activators. PMID- 10823242 TI - Nucleocytoplasmic shuttling and the control of NF-AT signaling. AB - The nuclear factors of activated T cells (NF-ATs) constitute a family of transcription factors that transduce calcium signals in the immune, cardiac, muscular and nervous systems. Like their distant relatives of the Rel family, including NF-kappaB, NF-ATs are cytoplasmic in resting cells and activated by means of induced nuclear import. Unlike NF-kappaB, however, NF-ATs show highly dynamic nuclear shuttling properties that have important implications for graded signaling by these molecules. This review focuses on recent advances in deciphering mechanisms by which calcium signaling regulates the nucleo cytoplasmic shuttling,and therefore transactivation functions of the NF-ATs. These discoveries highlight the interplay between nuclear import and export signals on NF-ATs, and the roles of the calcium-activated phosphatase calcineurin and NF-AT kinases in controlling the activity of these signals. They also reveal that NF-ATs, as well as other transcription factors controlled at the level of nuclear import, face the very real prospect of futile cycling across the nuclear envelope as a consequence of conflicting nuclear import and export signals. We discuss the molecular mechanisms by which calcineurin suppresses futile cycling, as well as the major challenges to our understanding of NF-AT signaling in diverse biological systems. PMID- 10823243 TI - Calcium and disease: molecular determinants of calcium crystal deposition diseases. AB - Deposition of basic calcium phosphate (hydroxyapatite, octacalcium phosphate and tricalcium phosphate) (BCP) and crystalline calcium pyrophosphate dihydrate (CPPD) is associated with a variety of aging-related pathologies, including osteoarthritis, cartilage degeneration and pseudogout. These diseases of calcium deposition serve as some of the best-studied examples of how calcium-regulated changes in gene expression can directly lead to pathogenic consequences. Tissue damage can result when crystals stimulate cells to release matrix-degrading molecules or secrete cytokines that stimulate the release of matrix-degrading molecules. Exposure of cultured cells to crystals induces expression of cellular proto-oncogenes such as c-fos, c-myc and c-jun, by a calcium-dependent mechanism, and this response can be blocked by a potential therapeutic compound, phosphocitrate. Activation of the c-fos and c-jun genes is directly involved in expression of metalloproteinases such as collagenase and stromelysin, suggesting that crystal-mediated activation of these genes is directly involved in pathogenesis. In this review recent advances in the molecular mechanisms responsible for crystal-mediated cell activation are discussed. PMID- 10823244 TI - Dysfunctional insulin secretion in type 2 diabetes: role of metabolic abnormalities. AB - Insulin secretion is finely tuned to the requirements of tissues by tight coupling to prevailing blood glucose levels. The normal regulation of insulin secretion is coupled to glucose metabolism in the pancreatic B cell, a major but not exclusive signal for secretion being closure of K+ ATP (adenosine' triphosphate)-dependent channels in the cell membrane through an increase in cytosolic ATP/adenosine diphosphate. Insulin secretion in type 2 diabetes is abnormal in several respects due to genetic causes but also due to the metabolic environment of the pancreatic B cells. This environment may be particularly important for the deterioration of insulin secretion which occurs with increasing duration of diabetes. Factors in the environment with potential importance include overstimulation, a negative effect of hyperglycemia per se ('glucotoxicity') as well as adverse effects of elevated fatty acids ('lipotoxicity'). Elucidating the mechanisms behind these factors as well as their clinical importance will pave the way for treatment which could preserve B cell function in type 2 diabetic patients. PMID- 10823246 TI - Heat shock effects on cell cycle progression. AB - In mammalian cells, short-term (acute) exposure to a moderate heat shock leads to a transient arrest of cells at mainly two cell cycle checkpoints, the G1/S and G2/M transitions. This is documented by the more or less synchronous resumption of cell cycle progression from these checkpoints during recovery. The reason for the accumulation of cells at these checkpoints may be found in activity thresholds of cyclin-dependent kinases (Cdks) at both transitions which are determined by (i) the amounts of the responsible cyclins, (ii) regulatory phosphorylation of the Cdks and (iii) the inhibition of Cdks by associated regulatory proteins (Ckis). All three regulatory systems may be subject to heat shock-dependent changes, the amounts of Ckis, in particular, being increased. Cdk dependent phosphorylation of the retinoblastoma protein and the subsequent release of active S-phase-specific transcription factors E2F/DP are considered as major heat-sensitive steps in cell cycle progression. Furthermore, high acute heat shock and long-term (chronic) heat treatment may lead to cell-type-specific forms of cell death. All types of responses to heat treatment are subject to adaptation after a 'priming' treatment, probably due to higher levels of heat shock proteins. PMID- 10823247 TI - Cell proliferation, carcinogenesis and diverse mechanisms of telomerase regulation. AB - Replication of linear genomes is incomplete and leaves terminal gaps. Solutions to this 'end replication' problem can be traced back to the prebiotic RNA world: 'fossils' of the presumptive archetypes of telomere structure and of the telomerase enzyme are retained in the terminal structures of some RNA viruses. Telomerase expression in mammals is ubiquitous in embryonic tissues but downregulated in somatic tissues of adults. Exceptions are regenerative tissues and, notably, tumor cells. Telomerase activation is controlled by cellular proliferation, and it is an early step in the development of many tumors. In contrast to mammals, indeterminately growing multicellular organisms, such as fish and crustaceae, maintain telomerase competence in all somatic tissues. In human tumor diagnostics, detection of proliferation markers with monoclonal antibodies is well established, and in this review, the significance of additional telomerase assays is evaluated. Telomerase inhibitors are attractive goals for application in tumor therapy, and telomerase knockout mice have proven that telomere erosion limits the lifespan of cells in vivo. In contrast, telomerase stimulation can be used to expand the potential of cellular proliferation in vitro, with possible applications for transplantation of in vitro expanded human cells, for immortalizing primary human cells as improved tissue models and for the isolation of otherwise intractable products, such as genuine human monoclonal antibodies. PMID- 10823245 TI - Immunoglobulin light chains, glycosaminoglycans, and amyloid. AB - Immunoglobulin light chains are the precursor proteins for fibrils that are formed during primary amyloidosis and in amyloidosis associated with multiple myeloma. As found for the approximately 20 currently described forms of focal, localized, or systemic amyloidoses, light chain-related fibrils extracted from physiological deposits are invariably associated with glycosaminoglycans, predominantly heparan sulfate. Other amyloid-related proteins are either structurally normal, such as beta2-microglobulin and islet amyloid polypeptide, fragments of normal proteins such as serum amyloid A protein or the precursor protein of the beta peptide involved in Alzheimer's disease, or are inherited forms of single amino acid variants of a normal protein such as found in the familial forms of amyloid associated with transthyretin. In contrast, the primary structures of light chains involved in fibril formation exhibit extensive mutational diversity rendering some proteins highly amyloidogenic and others non pathological. The interactions between light chains and glycosaminoglycans are also affected by amino acid variation and may influence the clinical course of disease by enhancing fibril stability and contributing to resistance to protease degradation. Relatively little is currently known about the mechanisms by which glycosaminoglycans interact with light chains and light-chain fibrils. It is probable that future studies of this uniquely diverse family of proteins will continue to shed light on the processes of amyloidosis, and contribute as well to a greater understanding of the normal physiological roles of glycosaminoglycans. PMID- 10823248 TI - Molecular modeling of mammalian cytochrome P450s. AB - The cytochrome P450s are a superfamily of hemoprotein enzymes responsible for the metabolism of a wide variety of xenobiotic and endogenous compounds. The individual P450s exhibit unique substrate specificity and stereoselectivity profiles which reflect corresponding differences in primary sequence and tertiary structure. In the absence of an experimental structure models for mammalian P450s have been generated by their homology with bacterial P450s of known structure. The rather low sequence identity between target and template proteins renders P450 modeling a challenging task. However, the substrate recognition properties of several P450s are consistent with recently developed working models. This review summarizes the major concepts and current approaches of molecular modeling of P450s. PMID- 10823249 TI - Inhibition of xanthine oxidase by liquiritigenin and isoliquiritigenin isolated from Sinofranchetia chinensis. AB - The methanol extract of the stem of Sinofranchetia inhibited the activity of xanthine oxidase in vitro. Bioassay-guided purification led to the isolation ofliquiritigenin and isoliquiritigenin as the main xanthine oxidase inhibitors. This inhibition of enzyme activity was found to be dose dependent, with an IC50 value of approximately 49.3 microM for liquiritigenin and 55.8 microM for isoliquiritigenin. Lineweaver-Burk transformation of the inhibition data indicated that the inhibition was of a mixed type for both liquiritigenin and isoliquiritigenin. For liquiritigenin, the Ki and K(I) were determined to be 14.0 microM and 151.6 microM, respectively. For isoliquiritigenin, the Ki and K(I) were determined to be 17.4 microM and 81.9 microM, respectively. These results suggest that these natural products could be used to treat conditions where the inhibition of xanthine oxidase is warranted. PMID- 10823250 TI - Immunomodulation by recombinant human interleukin-8 and its signal transduction pathways in invertebrate hemocytes. AB - We report the presence of interleukin (IL)-8-immunoreactive molecules in hemocytes from the mollusc Mytilus galloprovincialis. Functional studies demonstrate that recombinant human (rh)IL-8 provokes conformational changes, induces chemotaxis, and increases bacterial phagocytic activity in hemocytes. rhIL-8 induces cell shape changes via protein kinase A and C pathways. These morphological changes are followed by reorganization of the actin microfilaments. The findings suggest that, as previously reported for other cytokines, IL-8 is well conserved and deeply involved in immune functions from invertebrates to mammals. PMID- 10823251 TI - Resting potential of excitable neuroblastoma cells in weak magnetic fields. AB - The mechanism by which static and low-frequency magnetic fields are transduced into biological signals responsible for reported effects on brain electrical activity is not yet ascertained. To test the hypothesis that fields can cause a subthreshold change in the resting membrane potential of excitable cells, we measured changes in transmembrane current under voltage clamp produced in SH-SY5Y neuroblastoma cells, using the patch-clamp method in the whole-cell configuration. In separate experiments, cells were exposed to static fields of 1, 5, and 75 G, to time-varying fields of 1 and 5 G, and to combined static and time varying fields tuned for resonance of Na+, K+, Ca2+, or H+. To increase sensitivity, measurements were made on cells connected by gap junctions. For each cell, the effect of the field was evaluated on the basis of 100 trials consisting of a 5-s exposure immediately followed by a 5-s control period. In each experiment, the field had no discernible effect on the transmembrane current in the vicinity of zero current (- 50 mV voltage clamp). The sensitivity of the measuring system was such that we would have detected a current corresponding to a change in membrane potential as small as 38 microV. Consequently, if sensitivity of mammalian cells to magnetic fields is mediated by subthreshold changes in membrane potential, as in sensory transduction of sound, light, and other stimuli, then the ion channels responsible for the putative changes are probably present only in specialized sensory neurons or neuroepithelial cells. A change in transmembrane potential in response to magnetic fields is not a general property of excitable cells in culture. PMID- 10823252 TI - Recombinant factor VIIa is a pan-hemostatic agent? PMID- 10823253 TI - The endothelial cell protein C receptor. PMID- 10823254 TI - Use of recombinant factor VIIa in 3 patients with inherited type I Glanzmann's thrombasthenia undergoing invasive procedures. AB - The treatment of bleeds in Glanzmann's thrombasthenia is a challenging issue, especially when repeated platelet transfusions have induced anti-glycoprotein (GP) IIb-IIIa or anti-HLA allo-immunisation. In an attempt to find an alternative treatment regimen, we used recombinant factor VIIa (rFVIIa, NovoSeven, Novo Nordisk, Denmark) as first-line therapy in 3 patients with Glanzmann's thrombasthenia and anti-GPIIb-IIIa iso-antibodies who were scheduled for invasive procedures. The administration of an initial bolus dose of rFVIIa (70-110 microg/kg) was immediately followed by continuous infusion at the rate of 9-30 microg/kg/h for 3-15 days. The treatment resulted in an excellent clinical efficacy and tolerance in 2 cases. In the third patient, whereas efficacy was excellent at the surgical site, pharyngonasal bleeds of traumatic origin persisted for 10 days, and a severe thromboembolic complication occurred 5 days after discontinuation of rFVIIa. Complementary studies are needed for patients with congenital platelet disorders in order to evaluate the safety and the potential therapeutic place of rFVIIa treatment. PMID- 10823255 TI - Location and extent of deep vein thrombosis in patients with and without FV:R 506Q mutation. AB - Resistance to activated protein C due to FV:R 506Q mutation is the most common known genetic risk factor for deep leg vein thrombosis (DVT). The aim of this prospective study was to describe and compare the location and extent of DVT, reflected by a scoring system, in a group of patients with and without FV:R 506Q mutation. Of 247 consecutively included patients undergoing phlebography 105 had a DVT, 36 (35%) in the FV:R 506Q mutation group and 69 (65%) in the non-FV:R 506Q mutation group. Compared to the non-FV:R 506Q mutation group there was a significant increase in the incidence of DVT in the FV:R 506Q mutation group (p = 0.041, OR = 1.79 [1.02-3.15]), a significantly lower mean DVT score of the iliofemoral vein segments (p = 0.0081) and a significantly lower incidence of DVT in the iliofemoral vein segments (p = 0.007, OR = 10.6 [1.3-83.3]), 1/36 (2.8%) compared to 16/69 (23.2%). As controls 288 blood donors were included, with and without FV:R 506Q mutation and with no history of DVT in order to evaluate risk factors of DVT. The odds ratio of an iliofemoral DVT was 0.5 ([0.06-3.90), p = 0.50]) when FV:R 506Q mutation was present, compared to the control group, and at locations below the iliofemoral segments 5.28 ([3.01-9.28], p = less than 0.0001). Our findings provide the basis of a detailed phlebographic description and for the first time, to our best knowledge, shows a specific phlebographic pattern that may be linked to an inherited hypercoagulable state. PMID- 10823256 TI - Fixed-dose, body weight-independent subcutaneous LMW heparin versus adjusted dose unfractionated intravenous heparin in the initial treatment of proximal venous thrombosis. EASTERN Investigators. AB - BACKGROUND: Body weight-adjusted subcutaneous low-molecular-weight heparin (LMWH) has been proven to be at least as effective and safe as dose-adjusted intravenous unfractionated heparin (UFH) for the treatment of patients with venous thromboembolism. However, body weight-adjusted dosage of low-molecular-weight heparin may be cumbersome and could lead possibly to incorrect dosing. Therefore a fixed LMWH dose, independent of body-weight, might rationalize initial treatment for venous thromboembolism. METHODS: Patients with proven proximal deep vein thrombosis were randomly assigned to fixed dose subcutaneous LMWH Certoparin (8,000 anti-factor Xa U b.i.d.; 265 patients) or to adjusted dose i.v. UFH (273 patients) for 12 days. Vitamin K antagonists were started between day 3 and 7 and continued for up to 6 months. The primary outcome measure was a 30 percent or greater improvement in the Marder Score, as revealed by repeated venography on day 12 (end of the initial treatment). The secondary composite outcome measure included death, recurrent venous thromboembolism and major bleeding and was assessed at day 12 and after 6 months by a blinded adjunction committee. RESULTS: The Marder score improved by 30% or more in 30.3% and 25.0% of patients assigned to LMWH (198 paired venograms) and UFH (192 paired venograms), respectively (2p = 0.26). At the end of the initial treatment, the composite outcome was observed in 4 of the 265 patients (1.5%) randomized to LMWH, as compared with 14 of the 273 patients (5.1%) randomized to UFH (2p = 0.03). At 6 months these figures were 6.8% and 12.8%, respectively (risk reduction 0.53, confidence interval 0.31-0.90, 2p = 0.02). CONCLUSION: Fixed dose subcutaneous LMWH certoparin is at least as efficacious as UFH in resolving proximal vein thrombosis. PMID- 10823257 TI - Incidence of venous thromboembolism: a community-based study in Western France. EPI-GETBP Study Group. Groupe d'Etude de la Thrombose de Bretagne Occidentale. AB - The incidence of venous thromboembolism has been studied during one year in a defined population of 342,000 inhabitants. The overall incidence (95% confidence interval) of venous thromboembolism was found to be 1.83 per thousand per year (1.69 to 1.98). The incidences of deep venous thrombosis and pulmonary embolism were 1.24 per thousand per year (1.12 to 1.36) and 0.60 per thousand per year (0.52 to 0.69), respectively. The incidence of venous thromboembolism rose markedly with increasing age for both sexes; over the age of 75, the annual incidence reached 1 per 100. Sixty three percent of the patients were at home when venous thromboembolism occurred. Of these, sixteen percent had been previously hospitalised within three months. These results raise concerns on identification of medical patients at high risk and effective prophylaxis. PMID- 10823258 TI - A prospective controlled trial comparing weekly self-testing and self-dosing with the standard management of patients on stable oral anticoagulation. AB - Oral anticoagulant therapy requires frequent laboratory controls of its intensity to assure therapeutic efficacy and to prevent potentially life threatening adverse events. It is generally assumed, that increasing the frequency of testing would lead to a better control of anticoagulation. We tested this hypothesis in a prospective controlled trial comparing weekly self-testing and self-dosing (self management) with the standard-management of these patients in an anticoagulation clinic. Only patients with stable anticoagulation were included into the study. We recorded 2733 weekly determinations of the intensity of anticoagulation (INR) in 49 patients on self-testing and self-dosing and 539 determinations of the INR in 53 patients on standard-management. Two intensities of anticoagulation were used in each group: a target INR of 3.5 for patients with artificial heart valves (target range: 2.5-4.5) and a target INR 2.5 (target range: 2.0-3.0) for patients with atrial fibrillation or venous thromboembolism. The deviation from the target INR, the fraction of INR determinations within the preset therapeutic range and the difference between the target INR and the actually achieved mean INR were the three major endpoints of the study. The mean deviation from the target INR was smaller in the groups of patients on self-management compared to the patients on standard-management. Individual deviations were significantly (p <0.0001) dependent on the type of management in interaction with the treatment intensity in a general linear model. Patients on weekly self-testing and self-dosing had more INR values within the therapeutic range than patients on standard-management (86.2% vs. 80.1% at INR range 2.5-4.5; 82.2 vs. 68.9 at INR range 2.0-3.0). The achieved mean INR was almost identical with the target INR in the patients on self-management but was significantly (p <0.005) below the target INR in the high intensity anticoagulation group on standard-management (target INR:3.5; achieved mean INR: 3.19; CI 0.95: 3.05-3.34). Our data show, that weekly self-testing and self-dosing leads to a better control of anticoagulation than standard treatment in an anticoagulation clinic. PMID- 10823259 TI - Outcome and one year follow-up of intra-arterial staphylokinase in 191 patients with peripheral arterial occlusion. AB - Wild-type or equipotent variants of recombinant staphylokinase (rSak) were given intra-arterially (as a 2 mg bolus injection followed by an infusion of 1 mg/h or 0.5 mg/h overnight, with concomitant heparin [1000 IU/h]) to 191 patients of less than 80 years (62 +/- 1 years, mean +/- SEM), with a peripheral arterial occlusion (PAO) of less than 120 days (mean 14 +/- 1 days, median 11 days, 5 to 95 percentiles 3 to 30 days). Ninety nine patients presented with acute or subacute ischemia, 57 with severe claudication, 33 with chronic rest pain and 2 with gangrene. Occlusion occurred in 122 native arteries and in 69 grafts. Revascularization was complete in 83 percent (158/191), partial in 13 percent (24/191) and absent in 4 percent (7/191) after administration of 12 +/- 0.5 mg rSak over 14 +/- 0.7 h. Complete revascularization of acute occlusions of popliteal or more distal arteries was less frequent (60 percent, 15/25) than of acute occlusions of more proximal native arteries (95 percent, 37/39, p <0.001) or grafts (89 percent, 50/56, p = 0.005). Additional endovascular procedures were performed in 47 percent and subsequent elective bypass surgery in 23 percent of patients. Major bleeding occurred in 12 percent (23/191), one month mortality was 3.1 percent (6/191) and one year mortality was 6.9 percent (12/174). However, four patients (2.1 percent) had an intracranial bleeding following therapy: a 85 year old woman with severe diabetic arteriopathy, who was included in violation of the protocol, a 79 and a 74-year-old woman and a 74-year-old man, all with severe hypertension and limb threatening ischemia; these four patients died within two months after treatment. Amputations were performed within the first year in 16 of 162 surviving patients (9.8 percent): in 7 percent (7/96) with an occluded native artery and 14 percent (9/66) with an occluded graft (p = 0.19). No significant difference in lysis rate, one month mortality or one year amputation-free survival was observed in occlusions of recent onset (< or =14 days, n = 126) as compared to occlusions of longer duration (>14 days, n = 65). Treatment was interrupted prematurely in 4 patients because of a suspected allergic reaction. Fibrinogen levels remained unaffected during treatment (3.3 +/ 0.1 g/l before vs. 3.3 +/- 0.1 g/l after infusion, n = 167). In conclusion, rSak appears to be a highly effective thrombolytic agent in patients with PAO, resulting in a low one month mortality (3.1 percent) and a high one year amputation free survival (84 percent), with an acceptable incidence of major bleedings, but with occasional fatal intracranial hemorrhages. PMID- 10823260 TI - Improvement of walking distance by defibrotide in patients with intermittent claudication--results of a randomized, placebo-controlled study (the DICLIS study). Defibrotide Intermittent CLaudication Italian Study. AB - Defibrotide is an antithrombotic drug which enhances prostacyclin production and activates fibrinolytic system. The aim of this study was to investigate the improvement of walking distance in patients with intermittent claudication treated with defibrotide. DICLIS was a double blind, placebo-controlled study which included patients with walking distance autonomy at a standardized treadmill test < or =350 > or =100 meters. A total of 310 patients were randomly allocated to placebo (n = 101), defibrotide 800 mg/day (n = 104) or defibrotide 1200 mg/day (n = 105). During a one year follow-up, the Absolute Walking Distance (AWD) was measured six times (0, 30, 60, 90, 180, 360 days). Similar improvement in walking distance was found in the three groups until the 90th day; thereafter placebo group showed no further increase, while AWD continued to increase in the defibrotide groups. Between the 180th and 360th day visits, AWD was significantly higher (P <0.01) in patients given defibrotide than in patients given placebo. No difference in efficacy was observed between the two dosages of defibrotide. No differences in side effects were observed among the three groups. The results of the present trial suggest that long-term administration of defibrotide improves walking distance in patients with intermittent claudication. PMID- 10823261 TI - Hemostatic changes following the modified Fontan operation (total cavopulmonary connection). AB - Thromboembolism is a serious complication after Fontan operation, which may be caused by alterations of the coagulation system. We therefore investigated pro- and anticoagulant factors in 20 patients aged 4 to 21 years, 4 to 63 months following total cavopulmonary connection. Furthermore we compared markers of thrombin activation and fibrinolysis and in vitro clotting and clot-lysis to age matched healthy subjects. Compared to results of age-matched controls, the Fontan operated individuals had significant decreases in levels of protein C (0.88 U/ml in controls, 0.67 U/ml in patients; p <0.001) and protein S (1.05 in controls, 0.93 U/ml in patients; p <0.05). Moreover, half of the patients had high values of FVIII (>1.5 IU/ml), which are associated with an increased thrombotic risk. These changes may result in enhanced generation of thrombin and plasmin, indicated by our finding of increased thrombin-antithrombin III (TAT) and plasmin antiplasmin (PAP) levels and a similar trend in prothrombin fragments F1+2. Clot lysis tests, global coagulation tests, red blood cell count, liver enzymes AST, ALT, but not GGT, were generally within the normal ranges. PMID- 10823262 TI - Decreased sialic acid content of plasma von Willebrand factor in precapillary pulmonary hypertension. AB - In pulmonary hypertension, defective von Willebrand factor protein (vWF) lacking large multimers is present in circulation. This is associated with evidence of chronic endogenous platelet activation. Since asialo vWF has been shown to promote platelet activation and aggregation, we decided to investigate possible changes in the sialic acid content of plasma vWF in patients with precapillary pulmonary hypertension. vWF-associated sialic acid was measured indirectly as a wheat germ agglutinin-reactive substance (WGA-RS, Western blotting), and directly, as a thiobarbituric acid-reactive substance (TBA-RS, spectrophotometric reading). In the sixteen patients we studied (ages 8-45 yr), circulating vWF concentration was 2.18 times normal (p <0.001). However, patient vWF subunit contained 19% (WGA-RS) to 24% (TBA-RS) less sialic acid than the normal protein (p <0.05 for both determinations). In five patients, vWF-associated sialic acid was below 50% normal. We conclude that circulating vWF is hyposialylated in precapillary pulmonary hypertension and speculate that this might influence its interaction with platelets in vivo in these patients. PMID- 10823263 TI - Pravastatin: an antithrombotic effect independent of the cholesterol-lowering effect. AB - Lipid-lowering with statins reduces blood thrombogenicity. However, it is unknown whether this is purely due to LDL-cholesterol reduction, or it is related to a statin or agent specific effect. We investigated the relationship between reduction in blood thrombogenicity and the magnitude of low-density lipoprotein cholesterol (LDL-C) during pravastatin therapy. We prospectively followed for 6 months 57 hyperlipidemic patients who initiated therapy with pravastatin, and 36 patients who were randomized into placebo plus diet. Pravastatin-treated patients were grouped according to the LDL-C reduction at 6 months; (i) "adequate LDL-C reduction": LDL-C reduction >30% from baseline or LDL-C<125 mg/dl (n = 38; LDL-C reduction 74 +/- 4 mg/dl; 6-month LDL-C 119 +/- 5 mg/dl); (ii) "inadequate LDL-C reduction": neither of the above criteria (n = 19; LDL-C reduction 31 +/- 5 mg/dl; 6-month LDL-C 158 +/- 6 mg/dl). Placebo patients were divided into those "with LDL-C reduction" (n = 17, mean reduction 21 +/- 5 mg/dl) and those "without LDL reduction" (n = 19). The following parameters were altered at 6 months in both patients with "adequate" and "inadequate" LDL-C reduction: (1) tissue plasminogen activator decreased by 1.4 +/- 0.4 and 1.5 +/- 0.5 ng/ml respectively (p = NS); (2) plasminogen activator inhibitor-1 decreased by 8.7 +/- 2.0 and 10.1 +/- 2.7 ng/ml respectively (p = NS); (3) thrombus formation under dynamic flow conditions decreased by 3.5 +/- 0.9 and 2.8 +/- 1.2 microm2 x 10(3) respectively (p = NS). In contrast, no significant changes from baseline were noted in placebo treated patients, regardless of their LDL-C reduction category, and multivariate analysis eliminated LDL-C reduction as an independent predictor of reduction in thrombogenicity. Therefore, the reduction in thrombogenicity was not proportional to the magnitude of LDL-C reduction suggesting that a class or agent specific property is primarily responsible for the pro-fibrinolytic/antithrombotic effects observed. PMID- 10823264 TI - Gestational outcome in thrombophilic women with recurrent pregnancy loss treated by enoxaparin. AB - Inherited and acquired thrombophilia are associated with recurrent pregnancy loss (RPL). We have evaluated the efficacy and safety of the low molecular weight heparin enoxaparin in 50 women, (mean age 26 +/- 3 years) with RPL (> or =3 losses in 1st, > or =2 losses in 2nd and > or =1 loss in 3rd trimester) who were found to harbor thrombophilia. Twenty-seven had a solitary thrombophilic defect, and twenty-three women had combined thrombophilic defects: 17--two defects and 6- three defects. Following diagnosis of thrombophilia, sixty-one subsequent pregnancies were treated with the low molecular weight heparin enoxaparin throughout gestation until 4 weeks after delivery. Dosage was 40 mg/day in women with solitary defect and 80 mg/day in combined defects. Aspirin, 75 mg daily was given in addition to enoxaparin to women with antiphospholipid syndrome. Forty six out of 61 (75%) gestations treated by enoxaparin resulted in live birth compared to only 38/193 (20%) of the untreated pregnancies in these 50 women prior to diagnosis of thrombophilia (p <0.00001). In 23 women without a single living child following 82 untreated gestations, antithrombotic therapy resulted in 26/31 (84%) successful deliveries (p <0.0001). In 20 women with a prior living child, antithrombotic therapy improved successful delivery from 33/86 (38%) to 20/21 (95%) (p <0.0001). Enoxaparin dose of 40 mg/day resulted in live birth in 24/35 (69%) of gestations, compared to 19/23 (83%) gestations in women treated with 80 mg/day (p = 0.37). Only one thrombotic episode and one mild-bleeding episode were noticed during enoxaparin therapy. Enoxaparin is safe and effective in prevention of pregnancy loss in women with inherited and acquired thrombophilia. PMID- 10823265 TI - A comparison of point-of-care instruments designed for monitoring oral anticoagulation with standard laboratory methods. AB - Our study compared point-of-care (POC) device monitoring with traditional clinical laboratory methods device of patients on oral anticoagulant therapy. The POC devices used in the study were Coumatrak, CoaguChek, CoaguChek Plus, Thrombolytic Assessment System (TAS) PT-One, TAS PTNC, TAS PT, Hemachron Jr. Signature, ProTime Microcoagulation System, and Medtronics ACT II. The clinical laboratory method used thromboplastins with different ISI values: Innovin and Thromboplastin C Plus (TPC). All POC INRs showed strong correlation with both laboratory methods, with correlation coefficients of >0.900. All POC methods demonstrated a significant (p <0.05) difference in INR values, except the TAS PTNC and ACT II INRs (p: 0.12 and 0.71 respectively) when compared with Innovin INRs. All POC INRs were significantly different from TPC generated INRs (p <0.05). Comparisons of the POC INRs to the group mean of the POC methods, show higher correlation (R>0.93), but there were still significant (p<0.05) differences noted between the POC group INR mean and CoaguChek Plus, ACT II, TAS PT-One, TAS PTNC, and Hemachron Jr Signature INRs. These data indicate that POC INR biases exist between laboratory methods and POC devices. Until a suitable whole blood INR standardization method is available, we conclude that clinicians using point-of-care anticoagulation monitoring should be aware of differences between POC and parent laboratory values. PMID- 10823266 TI - The Lupus Ratio test--an interlaboratory study on the detection of Lupus anticoagulants by an APTT-based, integrated, and semi-quantitative test. Fifth International Survey of Lupus Anticoagulants--ISLA 5. AB - The Lupus Ratio (LR) test for lupus anticoagulants integrates screening, mixing with normal plasma and confirmation procedures into one assay. The sensitivity and reproducibility of the APTT based version of this assay was tested in an interlaboratory study that was part of the Fifth International Survey of Lupus Anticoagulants (ISLA-5). One LA negative plasma containing heparin, six LA positive plasmas and a normal pooled plasma (NP) were distributed to 31 laboratories world-wide together with two APTT reagents, one with a high and one with a low phospholipid concentration. The laboratories performed two APTTs, one with each reagent, on 1:1 mixtures of test plasma and NP. The ratio between the two clotting times was divided by the corresponding ratio for the NP. This final ratio is the LR of that plasma. The overall sensitivity was found to be 95.1%, and the normal, heparin-containing sample was reported to be negative by all the laboratories. When the results were grouped in low, medium and high positive plasmas, a "consensus" regarding the strength of each plasma was easily found. 85.0% of the results were in agreement with this consensus. This study shows that with the LR test, it is possible to obtain high interlaboratory agreement regarding the presence or absence of LA as well as the semi-quantification of this inhibitor. PMID- 10823267 TI - High molecular weight kininogen is cleaved by FXIa at three sites: Arg409-Arg410, Lys502-Thr503 and Lys325-Lys326. AB - We investigated the cleavage of high molecular weight kininogen (HK) by activated coagulation factor XI (FXIa) in vitro. Incubation of HK with FXIa resulted in the generation of cleavage products which were subjected to SDS-Page and analyzed by silverstaining, ligand-blotting and immunoblotting, respectively. Upon incubation with FXIa, bands were generated at 111, 100, 88 kDa on nonreduced and at 76, 62 and 51 kDa on reduced gels. Amino acid sequence analysis of the reaction mixtures revealed three cleavage sites at Arg409-Arg410, at Lys502-Thr503 and at Lys325 Lys326. Analysis of HK-samples incubated with FXIa for 3 min, 10 min and 120 min indicated HK to be cleaved first at Arg409-Arg410, followed by cleavage at Lys502 Thr503 and then at Lys325-Lys326. In conclusion, HK is cleaved by FXIa at three sites. Cleavage of HK by FXIa results in the loss of the surface binding site of HK, which may constitute a mechanism of inactivation of HK and of control of contact system activation. PMID- 10823268 TI - Major structural defects in the antithrombin gene in four families with type I antithrombin deficiency--partial/complete deletions and rearrangement of the antithrombin gene. AB - The molecular basis of quantitative antithrombin deficiency was investigated in four families predicted to have major antithrombin gene rearrangements. A 1,442 bp deletion and insertion of the sequence 5'T(n = 38-40)GAGACG was characterised in one case. Sequence surrounding the breakpoints contained two perfect, and one imperfect, inverted repeats which may have mediated formation of a stem loop structure on one strand during DNA replication potentiating the deletion. A 9,219 bp deletion spanning introns 2 to 5 was identified in a second family. The identical 6 bp sequence was upstream of each breakpoint and the 5' breakpoint was located in a sequence of the Alu 3 repeat predicted to be susceptible to strand breakage during transcription. This may have promoted misalignment, and deletion, of one of the repeats and the intervening DNA. A novel 1.8 kb antithrombin gene fragment was present in DNA digests from affected members of the third family suggesting a partial antithrombin gene duplication event while in the remaining family, evidence supporting a complete gene deletion was obtained. PMID- 10823269 TI - Protective effects of imidapril on He-Ne laser-induced thrombosis in cerebral blood vessels of stroke-prone spontaneously hypertensive rats. AB - Inhibitors of angiotensin converting enzyme (ACE) have been developed recently for therapeutic purposes in hypertension and ischemic cardiovascular diseases. Ogiku et al. reported that one such inhibitor, imidapril, significantly prolonged survival in stroke-prone spontaneously hypertensive rats (SHRSP). The present study was designed to investigate the effect of imidapril on cerebral blood vessels in SHRSP to clarify role of the ACE inhibitor in mechanisms of cerebral thrombosis and stroke. Imidapril was administered orally at 1.0 and 5.0 mg/kg/day for 3 weeks from the age of 7 weeks, and was shown to prevent the usual increase in blood pressure seen in these animals. It also delayed He-Ne laser-induced cerebral thrombosis and increased significantly the plasma concentration of nitric oxide metabolites (NO2/NO3). To confirm the association between nitric oxide (NO) and these effects of imidapril, an inhibitor of nitric oxide synthase, N(G)-nitro-L-arginine methyl ester hydrochloride (L-NAME) was dissolved in drinking water and administered to the animals for 3 weeks. Four of six rats died from stroke when L-NAME was given alone. When imidapril (5.0 mg/kg/day) was administered with L-NAME, however, the animals showed no signs or symptoms of stroke. In these instances, therefore, the concurrent administration of L-NAME with imidapril reversed significantly the effects of imidapril. Intravenous injection of imidaprilat (100 microg/kg), an active metabolite of imidapril, also decreased blood pressure significantly and increased the plasma levels of NO2/NO3 after 5 min. Moreover, imidaprilat enlarged arteriolar diameters and caused an increase in red cell velocity and mean blood flow in pial arterioles after 15 min. The results strongly suggested that imidapril protects cerebral vessels in SHRSP by elevating the release of NO, thereby improving the cerebral circulation and reducing the tendency to thrombosis and stroke. PMID- 10823270 TI - Detection of complement-fixing antiphospholipid antibodies in association with thrombosis. AB - Antiphospholipid antibody (aPL) is a hallmark of antiphospholipid syndrome (APS), characterized by thrombosis and recurrent fetal loss. We developed a novel ELISA system to detect complement-fixing ability of anticardiolipin antibody (aCL), and evaluated its clinical usefulness through studying the prevalence of the antibodies in rheumatic diseases, especially in association with thrombosis and recurrent abortion. Among 189 patients with rheumatic diseases, the complement fixing aCL was positive in 26 (83.9%) of 31 patients with APS and 2 (1.3%) of 158 with other disease categories, whereas it was not positive among 52 normal subjects. Twenty-seven of 28 patients (96.4%), who were positive for complement fixing aCL, had the episodes or history of thrombosis and/or recurrent abortion, at the time we studied. The remaining one in this group developed APS manifesting pulmonary infarction and occlusion of mesenteric artery 6 months after the evaluation. The sensitivity and specificity of this assay system were 75.0% and 99.3%, respectively, in relation with thrombotic episodes. On the other hand, the IgG aCL were positive in 28 (77.8%) of 36 cases with recent thrombotic episodes and 24 (15.7%) of 153 cases with no recent thrombotic episodes. The sensitivity and specificity of IgG aCL assay system were 77.8% and 84.3%, respectively, in relation with thrombotic episodes. These results indicate that complement-fixing aCL may specifically occur in association with the episodes of thrombosis and/or recurrent abortion in patients with APS compared to IgG-aCL. The method for detecting the complement-fixing aCL is simple, and it provides the useful diagnostic marker for thrombotic manifestations associated with APS. PMID- 10823271 TI - Cloning and characterization of the murine coagulation factor X gene. AB - The gene encoding murine coagulation factor X (fX) was isolated and characterized from a lamdaFIX II library generated from murine genomic DNA. The 20130 bp sequence contains 18049 nucleotides that extend from the initiating methionine to the polyadenylation site. 1056 nucleotides 5' of the start codon were determined and contain putative start sites for the FX mRNA as well as sites for binding of putative transcription factors. The sequence extends 1024 3' of the polyadenylation site. The gene contains 8 exons and 7 introns which were determined by comparing the mouse FX cDNA and gene sequences. The exonic structure of the gene is similar to that of the other mammalian vitamin K dependent serine proteases of the coagulation system. These include an exon encoding the prepropepetide, the gla-domain, a short helical stack, two exons for the two EGF domains, the activation pepetide, and two exons encoding the serine protease domain. The 5' sequence of the mouse FX gene overlaps with the 3' region of the FVII gene indicating that the murine FVII and FX gene are arranged in a head to tail arrangement as they are in humans. PMID- 10823272 TI - Fibrinogen and fibrin protect fibroblast growth factor-2 from proteolytic degradation. AB - We have recently reported that fibrinogen and fibrin bind to fibroblast growth factor-2 (FGF-2) and potentiate its ability to stimulate proliferation of endothelial cells. In the present report, we have investigated the potential of fibrinogen and fibrin to protect FGF-2 from proteolytic degradation. FGF-2 was incubated with trypsin or chymotrypsin in the presence or absence of fibrinogen or fibrin and proteolysis of FGF-2 was assessed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. In the absence of fibrinogen there was progressive tryptic degradation of FGF-2, but in the presence of fibrinogen, FGF-2 was completely protected from trypsin with no evidence of degradation. The degree of protection was maximum at a molar ratio of FGF-2 to fibrinogen 1:2. Fibrinogen afforded similar protection from degradation by chymotrypsin. Polymerized fibrin provided partial protection of FGF-2 from tryptic degradation, with intact FGF-2 present for up to 360 min. Fibrin provided nearly complete protection from chymotrypsin. These observations indicate that binding of FGF-2 to fibrinogen or fibrin provides protection from proteolytic degradation, and this may modulate its cell proliferative activity. PMID- 10823273 TI - Vitronectin and substitution of a beta-strand 5A lysine residue potentiate activity-neutralization of PA inhibitor-1 by monoclonal antibodies against alpha helix F. AB - Some monoclonal antibodies against plasminogen activator inhibitor-1 (PAI-1) are able to inhibit its reaction with its target proteinases. We have characterized the effect on PAI-1 of two monoclonal antibodies, Mab-2 and Mab-6, with overlapping epitopes in a sequence encompassing beta-strand 1A, alpha-helix F, and the loop connecting alpha-helix F and beta-strand 3A (the hF/s3A loop). Mab-2 reduced the inhibitory activity of wild type PAI-1 and almost totally abolished the inhibitory activity of a PAI-1 variant harboring an Ala substitution of Lys 325 (335 in the alpha1-proteinase inhibitor template residue numbering) in beta strand 5A. In both cases, the neutralizing effect of the antibody was strongly potentiated by vitronectin. Mab-6 had no effect on wild type PAI-1, but reduced the inhibitory activity of the K325A variant. The effect of Mab-6 was not potentiated by vitronectin. With both Mab-2 and Mab-6, the neutralization of PAI 1 activity was associated with PAI-1 substrate behaviour. Mab-2, but not Mab-6, prevented vitronectin from rescuing PAI-1 from cold-induced substrate behaviour. We propose that the antibodies act by weakening the anchoring of alpha-helix F to the adjacent structures, resulting in an increased flexibility of beta-strand 5A and the hF/s3A loop and a changed conformational response to the binding of vitronectin in the alpha-helix E region. The potentiating effect of vitronectin on neutralization of PAI-1 by antibodies is a novel concept in the development of compounds for neutralizing PAI-1 in vivo. PMID- 10823274 TI - Influence of SIN-1 on platelet Ca2+ handling in patients with suspected coronary artery disease: ex vivo and in vitro studies. AB - The 3-morpholinosydnonimine (SIN-1) generates both nitric oxide (NO) and superoxide anion (O2-). It elicits dose-dependent vasodilation in vivo, in spite of the opposite effects of its breakdown products on vascular tone and platelet aggregation. This study was designed to investigate the influence of intravenous SIN-1 injection on platelet Ca2+ handling in patients undergoing coronary angiography. SIN-1 administration reduced cytosolic [Ca2+] in unstimulated platelets by decreasing Ca2+ influx. It attenuated Ca2+ mobilization from internal stores evoked by thrombin or thapsigargin. In vitro studies were used as an approach to investigate how simultaneous productions of NO and O2- from SIN-1 modify thrombin- or thapsigargin-induced platelet Ca2+ mobilization. Superoxide dismutase, the O2- scavenger, enhanced the capacity of SIN-1 to inhibit Ca2+ mobilization but catalase had no effect. This suggests that the effects of SIN-1 on platelet Ca2+ handling resemble those of NO, but are modulated by simultaneous O2- release, independently of H2O2 formation. PMID- 10823276 TI - Involvement of activated integrin alpha2beta1 in the firm adhesion of platelets onto a surface of immobilized collagen under flow conditions. AB - Recently, we demonstrated that agonist-induced activation of the platelet surface collagen-receptor integrin alpha1beta2 converts it to an active form that can bind soluble collagen with high affinity (Jung, SM, Moroi, M: J Biol Chem 1998; 273: 14827-37). Here, the involvement of alpha2beta1 activation and the high affinity binding property of activated alpha2beta1 in platelet adhesion to a collagen surface under flow conditions were analyzed. Platelet adhesion to immobilized collagen was measured in the presence of TS2/16, an activating anti integrin alpha2beta1 antibody, and inhibiting antibodies, Gi9 and 6F1. TS2/16 decreased the moving velocity of platelets on the collagen surface, but Gi9 and 6F1 increased it, indicating that alpha2beta1 activation induces the tight binding of platelets to immobilized collagen under flow. Platelet adhesion, expressed as the surface area occupied by adhered platelets, in the presence of TS2/16 was similar to that in its absence. In contrast, adding Gi9 or 6F1 caused biphasic adhesion composed of a first phase, a lag phase whose length differed in each experiment, and a second phase adhesion with a rate similar to that of the control. This biphasic adhesion indicates that alpha2beta1 activity is inhibited and also suggests that some other factor(s) may contribute to the adhesion under flow. At concentrations where neither 6F1 nor Gi9 affected collagen-induced aggregation, these antibodies inhibited soluble collagen binding to thrombin activated platelets. Only at much higher concentration did 6F1 inhibit collagen induced aggregation. TS2/16 had no effect on the aggregation. The present results are evidence against the major involvement of integrin alpha2beta1 in platelet aggregation; instead, they indicate that integrin alpha2beta1 would be mainly associated with the tight binding of platelets to collagen. PMID- 10823275 TI - Thromboxane synthase has the same pattern of expression as platelet specific glycoproteins during human megakaryocyte differentiation. AB - Regulation of the platelet formation process is poorly understood. It has been shown that p45NF-E2 deficient mice have a profound defect in platelet formation and recently the first platelet/megakaryocytic gene regulated by NF-E2, thromboxane synthase (TXS), has been identified. In this study, we investigated TXS expression as a model of a gene regulated by NF-E2 during MK differentiation. Megakaryocytic cells derived from blood CD34+ cells were purified according to their stage of maturation on the basis of expression of CD34, CD41a and CD42a, permitting to define different stages in MK differentiation. By means of real time quantitative RT-PCR, we could determine that the level of TXS increased during differentiation in parallel with the expression of c-mpl and GPIIb (CD41). However, amounts of TXS transcripts increased about 1.6-fold more than that of GPIIb or c-mpl transcripts during maturation. Expression of TXS and MK specific proteins such as CD41a, CD42a and vWF was also correlated in maturing MKs. In addition, staining by anti-TXS antibody of proplatelet bearing MKs was not increased in comparison to that observed in mature MK, suggesting that TXS is not upregulated during platelet formation. In addition, we investigated whether TXS and cyclooxygenase could be involved in platelet formation by adding aspirin into the cultures. No significant decrease of platelet production was observed. In conclusion, this study shows that TXS is coordinately expressed with the other platelet proteins during MK differentiation but is not directly involved in platelet formation. PMID- 10823278 TI - Fast decrease of bleeding time by tranexamic acid in uremia. PMID- 10823277 TI - Mechanisms involved in the antiplatelet activity of Staphylococcus aureus lipoteichoic acid in human platelets. AB - In this study, gram-positive Staphylococcus aureus lipoteichoic acid (LTA) dose dependently (0.1-1.0 microg/ml) and time-dependently (10-60 min) inhibited platelet aggregation in human platelets stimulated by agonists. LTA also dose dependently inhibited phosphoinositide breakdown and intracellular Ca+2 mobilization in human platelets stimulated by collagen. LTA (0.5 and 1.0 microg/ml) also significantly inhibited thromboxane A2 formation stimulated by collagen in human platelets. Moreover, LTA (0.1-1.0 microg/ml) dose-dependently decreased the fluorescence of platelet membranes tagged with diphenylhexatrience. Rapid phosphorylation of a platelet protein of Mr. 47,000 (P47), a marker of protein kinase C activation, was triggered by PDBu (30 nM). This phosphorylation was markedly inhibited by LTA (0.5 and 1.0 microg/ml) within a 10-min incubation period. These results indicate that the antiplatelet activity of LTA may be involved in the following pathways: LTA's effects may initially be due to induction of conformational changes in the platelet membrane, leading to a change in the activity of phospholipase C, and subsequent inhibition of phosphoinositide breakdown and thromboxane A2 formation, thereby leading to inhibition of both intracellular Ca+2 mobilization and phosphorylation of P47 protein. Therefore, LTA-mediated alteration of platelet function may contribute to bleeding diathesis in gram-positive septicemic and endotoxemic patients. PMID- 10823279 TI - Type I CD36 deficiency in humans is not associated with insulin resistance syndrome. PMID- 10823280 TI - The TFPI 536C-->T mutation is not associated with increased risk for venous or arterial thrombosis. PMID- 10823281 TI - Is clinical probability assessment of deep vein thrombosis by a score really standardized? PMID- 10823282 TI - New recombinant standard for FVIII concentrate gives same results as previous plasma derived standards on a range of FVIII products. PMID- 10823283 TI - Genotyping of the stromelysin-1 5A/6A promoter polymorphism using a heteroduplex generator. PMID- 10823284 TI - No evidence for enhanced thrombin formation through displacement of annexin V by antiphospholipid antibodies. PMID- 10823286 TI - Quantitation of serotonin from platelets using enzyme-immunoassay in the presence of serum from HIT-patients. PMID- 10823285 TI - Miconazole oral gel potentiates warfarin anticoagulant activity. PMID- 10823287 TI - Lack of association of the prothrombin gene variant G20210A with myocardial infarction in Caucasian males. PMID- 10823288 TI - Influence of the nt 2148 A to G substitution (Pro 626 dimorphism) in the PROS1 gene on circulating free protein S levels in healthy volunteers--reappraisal of protein S normal ranges. PMID- 10823289 TI - Phenprocoumon-induced hepatitis in a patient with a combined hereditary hemostatic disorder. PMID- 10823290 TI - Plastic versus siliconized glass tubes: evaluation in current laboratory practice. PMID- 10823291 TI - How effective is long-term lithium prophylaxis? PMID- 10823292 TI - Does increasing dose improve efficacy in patients with poor antidepressant response: a review. AB - OBJECTIVE: Therapeutic strategies in depressed patients with no or partial response to adequate first-line antidepressant medication remain a matter of concern. This paper focuses on the strategy of dose increase. METHOD: This review was based on a systematic Medline search of papers dealing with antidepressant dose issues in major depression since the 1960s. RESULTS: The strategy of dose increase is poorly studied in clinical trials. CONCLUSION: Until this strategy is better studied, caution is advised in its use. However, antidepressants for which this strategy seems to be the most relevant are tricyclic drugs and serotonin and noradrenaline reuptake inhibitors. These results are discussed both in terms of therapeutic strategies for the clinician and in terms of clinical research. PMID- 10823293 TI - Lithium monitoring before and after the distribution of clinical practice guidelines. AB - OBJECTIVE: To determine whether distribution of clinical practice guidelines improves lithium monitoring and whether standards of monitoring differed between patients in psychiatric contact and those seen only in primary care. METHOD: Standards of monitoring were assessed for patients on lithium in northeast Scotland throughout 1995 and/or throughout 1996. Guidelines were circulated in January 1996 to all local general practitioners and psychiatrists. Monitoring was compared between 1995 and 1996 and for patients with and without psychiatric contact. RESULTS: Both primary care and psychiatric records were scrutinized for 422 and 403 patients prescribed lithium throughout 1995 and 1996, respectively. While monitoring was poor on several parameters during both years, frequency of measurement of both thyroid and renal function improved in 1996. Standards of monitoring were better for patients in psychiatric care. CONCLUSION: Standards of lithium monitoring require further improvement. Locally agreed practice guidelines are helpful but patients on lithium should be in continuing contact with an experienced psychiatrist. PMID- 10823294 TI - Therapeutic drug monitoring of selective serotonin reuptake inhibitors influences clinical dosing strategies and reduces drug costs in depressed elderly patients. AB - OBJECTIVE: This study was initiated in order to describe and evaluate the effects of a therapeutic drug monitoring (TDM) routine of selective serotonin reuptake inhibitors (SSRIs) on treatment strategies and drug costs in depressed elderly patients. METHOD: Blood samples were drawn from elderly depressed patients and analysed for steady-state trough serum concentrations of citalopram (n = 48), paroxetine (n = 48) or sertraline (n = 39). A global efficacy evaluation was made at baseline and after 6-9 months. Antidepressant drug costs before and after TDM were estimated. RESULTS: Eight samples were excluded due to technical problems or noncompliance. In 65 of the 127 (51.2%) remaining cases, the treatment strategy was changed according to the TDM outcome, in most a reduction of the prescribed dose. Bioanalytical TDM costs included the antidepressant drug costs after TDM were reduced by 10.2%. CONCLUSION: The results support the utility of TDM in the search for the individual minimum effective SSRI dose in the elderly. PMID- 10823295 TI - European Consultation-Liaison Psychiatric Services: the ECLW Collaborative Study. AB - OBJECTIVE: To describe the patterns of organization of consultation-liaison (C-L) services in 11 European countries in relation to hospital characteristics and national approaches to C-L psychiatry. METHOD: Cross-sectional survey. RESULTS: Services can best be described in terms of their size and seniority of their staff and whether or not they are multidisciplinary. Single-discipline services are based upon the standard medical consultant model, whereas those with multidisciplinary teams work in a way that is comparable with community mental health teams. German psychosomatic C-L services belonged to either model. National differences were found. CONCLUSION: This first international study provides empirical evidence for the wide variation in the organization of C-L services. In view of the increasing numbers of patients with psychiatric disorder who are being treated in general hospitals and the changing patterns of medical care there are important implications for clarification and improvement of the role of C-L services. PMID- 10823296 TI - Predictors for readmission risk of new patients: the Nordic Comparative Study on Sectorized Psychiatry. AB - OBJECTIVE: Predictors for readmission risk were investigated in this study, which forms part of the Nordic Comparative Study on Sectorized Psychiatry. METHOD: Included were a total of 837 consecutive 'new' patients (not in contact with the psychiatric services for at least 18 months) admitted to in-patient stay during a period of 1 year to seven psychiatric hospitals in four Nordic countries. RESULTS: Multivariate survival analyses showed that younger age predicted increased readmission risk. Stratifying on gender, diagnostic group and sector revealed a general pattern concerning age which was the only consistent main effect. Living alone and unemployed increased readmission risk in the non psychosis group, while receipt of aftercare decreased readmission risk in the psychosis group. A curvilinear relationship was found between availability of psychiatric resources and readmission risk: an intermediate amount of resources was associated with increased risk. CONCLUSION: Our findings support a hypothesis that readmission risk is multifactorially determined and that interactions have to be considered. PMID- 10823297 TI - Effect of number of episodes on wellbeing and functioning of patients with bipolar disorder. AB - OBJECTIVE: To examine the relationship between number of episodes and inter episode functioning in bipolar disorder. METHOD: Sixty-four euthymic subjects with bipolar affective disorder completed the Medical Outcomes Questionnaire Short Form and the Global Assessment of Functioning Scale. Goodness-of-fit models were used to define the relation between episode number and level of function. RESULTS: Non-linear logarithmic and power relations best described the association between number of episodes and outcome. Number of past depressions was a stronger determinant of outcome than past manias. CONCLUSION: Strategies to minimize the number of episodes experienced by patients with bipolar illness must be pursued aggressively if function is to be maintained, with particular attention given to minimizing episodes of depression. PMID- 10823298 TI - Excess cancer mortality in Western Australian psychiatric patients due to higher case fatality rates. AB - OBJECTIVE: To investigate the association between mental illness and cancer incidence, mortality and case fatality. METHOD: A population-based record linkage study was undertaken based on 172,932 patients of mental health services in Western Australia. Records of mental health service contacts were linked with cancer registrations and death records. RESULTS: While there was little difference in overall cancer incidence rates between psychiatric patients and the general community (RR in males 1.05, 95%, CI 1.02-1.09, RR in females 1.02, 0.98 1.05), cancer mortality was 39%, higher in males (95%) CI: 32-46%) and 24% higher in females (17-32%). CONCLUSION: People with mental illness in Western Australia do not show an increased incidence rate of cancer, but do have higher cancer mortality. This was attributed to a higher cancer case fatality rate among people with mental illness. PMID- 10823299 TI - Quantitative and qualitative aspects of obsessive-compulsive behaviour in children with attention-deficit hyperactivity disorder compared with tic disorder. AB - OBJECTIVE: Within the framework of associated psychopathology in child psychiatric disorders, this study focused on quantitative and qualitative aspects of obsessive-compulsive behaviour (OCB) in both attention-deficit hyperactivity disorder (ADHD) and chronic tic disorder/Tourette's disorder (TD). METHOD: Forty two healthy controls, 41 children with ADHD and 38 children with TD, aged 9-13 years, were investigated using the Leyton Obsessional Inventory--Child Version (LOI-CV), the Child Behaviour Checklist (CBCL) and an expert-rated structured parent interview to reflect a cross-informant view of OCB. RESULTS: Unexpectedly, self-reports of children with ADHD rather than children with TD showed the highest OCB scores in the LOI-CV. Qualitatively, ADHD-related OCB focused on the item subsets concerning 'dirt and contamination', 'repetition', 'overconscientiousness', and 'hoarding'. In the parent-rated CBCL, similar levels of OCB were reported for ADHD and TD patients. In contrast, only children with TD showed clinically relevant OCB according to expert ratings. CONCLUSION: Not only young TD patients but also children with ADHD should be investigated and monitored carefully for quantitative and qualitative aspects of OCB comorbidity. PMID- 10823300 TI - Suicide attempts and impulse control disorder are related to low cerebrospinal fluid 5-HIAA in mentally disordered violent offenders. AB - OBJECTIVE: The aim of this study was to investigate the relationship between low cerebrospinal fluid 5-hydroxyindoleacetic acid (CSF 5-HIAA) and aggressive acts in mentally disordered violent offenders. METHOD: CSF concentrations of 5 hydroxyindoleacetic acid (5-HIAA), homovanillic acid and 4-hydroxy-3 methoxyphenyl glycol were measured in 35 drug-free men convicted of homicide and currently undergoing forensic psychiatric investigation in a high security hospital. RESULTS: The mean metabolite concentrations in the homicide offenders did not differ from those of age- and body-height matched male control subjects. One-third of the murderers had made one or several suicide attempts, and their mean concentration of spinal fluid 5-HIAA was significantly lower than that of the remaining murderers. Subjects with impulse control disorder also had lower mean CSF 5-HIAA. A consistent pattern of higher CSF 5-HIAA in subjects with more self-reported anxiety traits was observed. CONCLUSION: The association between serotonin and suicidal behaviour holds for yet another subject group. Determining CSF 5-HIAA might be worth while in the routine assessment of psychiatrically ill homicide offenders. PMID- 10823301 TI - Low novelty-seeking differentiates obsessive-compulsive disorder from major depression. AB - OBJECTIVE: To make a direct comparison of patients with obsessive-compulsive disorder (OCD) and major depression (MD) and a normal control group in terms of the Temperament and Character Inventory (TCI) personality dimensions. METHOD: Additionally to 43 patients with primary OCD, 43 MD patients and 43 normal subjects who were matched against the OCD patients for sex and age filled out the TCI. RESULTS: Compared to the controls, the OCD and MD patients scored significantly higher on harm avoidance and significantly lower on self directedness and co-operativeness. The OCD patients scored significantly lower on novelty-seeking than the MD patients and the controls. CONCLUSION: Whereas OCD and MD share similar personality deviations on harm avoidance, self-directedness and co-operativeness, OCD is distinguishable from MD in terms of low novelty seeking. Low novelty-seeking may have a profound relationship to the specific aetiology of OCD. PMID- 10823302 TI - Systemic lupus erythematosus presenting as mania. AB - OBJECTIVE: Psychiatric manifestations of systemic lupus erythematosus (SLE) are well recognized but usually occur in the later stages of the illness, with organic syndromes being the most common. This case highlights the fact that SLE can present with mania. METHOD: Single case report. RESULTS: A 32-year-old woman was admitted to hospital with her first manic illness. Physical examination and investigations revealed her to be suffering from SLE although this had not been previously diagnosed. Her physical and psychiatric symptoms settled rapidly following treatment with prednisolone and chlorpromazine. CONCLUSION: The case emphasizes the polymorphous manifestations of SLE and the importance of considering physical disorders in patients with a first episode of psychosis. PMID- 10823303 TI - Clozapine-induced urinary incontinence: facts or artefacts? PMID- 10823304 TI - Amisulpride: its role in the therapeutic management of the schizophrenic patient. Introduction. PMID- 10823305 TI - Early diagnosis of schizophrenia--the first step towards secondary prevention. AB - OBJECTIVE: To review current knowledge of risk factors associated with schizophrenia and the development of diagnostic markers aimed at the delay or attenuation of the first psychotic episode. METHOD: Studies relating to the development of disease and the risk factors that could inform predictive markers are reviewed, including high-risk studies, birth-cohort studies, and retrospective and follow-back studies. RESULTS: Future schizophrenic patients present with delayed developmental milestones, speech and behavioural difficulties, and lower IQ scores than non-cases. CONCLUSION: Results are consistent with the notion that schizophrenia is a developmental disease and that such antecedents are present; further research is required to improve the specificity of diagnostic markers and predictive power before these can be used to prevent or delay psychotic episodes of schizophrenia. PMID- 10823306 TI - Cognitive impairment in schizophrenia: its impact on social functioning. AB - OBJECTIVE: To examine cognitive impairment in schizophrenia and its impact on social functioning. METHOD: Cohort studies recording cognitive and social development, studies of predictors of poor community outcome, and evidence from brain imaging studies are reviewed. RESULTS: Executive function deficits, and poor performance in verbal memory, vigilance, and working memory tests are strong predictors of poor community outcome and impairment in skills learning. PET scans of regional cerebral blood flow suggest a dynamic imbalance between different cerebral areas rather than overall loss of brain function. CONCLUSION: Chronic cognitive impairment is the strongest predictor of social disability. Results suggest that the impairment of brain function associated with executive deficits is not necessarily irreversible and may therefore be treatable with appropriate drug therapy. PMID- 10823307 TI - Amisulpride: a review of its efficacy in schizophrenia. AB - OBJECTIVE: To assess the efficacy of the new atypical antipsychotic drug, amisulpride. METHOD: Studies comparing the efficacy of amisulpride with that of haloperidol and risperidone, respectively, are reviewed. Outcome measures were Clinical Global Impression, Brief Psychiatric Rating Scale (BPRS), and Positive And Negative Symptom Scale (PANSS) scores. RESULTS: Amisulpride was at least as effective as haloperidol and risperidone in the improvement of positive symptoms, and significantly more efficacious than haloperidol in reducing PANSS negative subscores (P=0.038) in patients with acute exacerbations. Amisulpride demonstrated a greater improvement in BPRS total scores (P<0.05) and PANSS negative subscores (P=0.0001) than haloperidol after 12 months of treatment in chronic schizophrenic patients with acute exacerbations. CONCLUSION: Amisulpride can thus be considered for use as first-line treatment of acute and chronic schizophrenia. PMID- 10823308 TI - Safety profile of amisulpride in short- and long-term use. AB - OBJECTIVE: To assess the safety of the new atypical antipsychotic drug, amisulpride, in short- and long-term use. METHOD: Studies comparing the safety of amisulpride with that of haloperidol and risperidone, respectively, are reviewed. Safety was monitored by open adverse event reporting, the Simpson-Angus Scale, the Barnes Akathisia Scale and the Abnormal Involuntary Movement scale. RESULTS: In short- and long-term studies, amisulpride induced significantly less EPS and akathisia than haloperidol. Safety ratings were similar to risperidone in short term studies. In studies of chronic schizophrenia with predominant negative symptoms, amisulpride was similar to placebo. Endocrine effects were similar in amisulpride-, haloperidol- and risperidone-treated patients. Weight gain with amisulpride was significantly less than risperidone in a short-term study. No clinically important effects on haematological, hepatic or cardiac function were recorded. Data obtained in short- and long-term studies have been confirmed in extensive post-marketing surveillance data. CONCLUSION: Amisulpride has a broad spectrum of efficacy in schizophrenia without introducing the iatrogenic consequences associated with older therapies. PMID- 10823309 TI - Amisulpride: its role in the therapeutic management of the schizophrenia patient. Conclusions. PMID- 10823310 TI - Low-dose broad-band UVA in morphea using a new method for evaluation. AB - Until recently, various therapies for localized scleroderma have been used with limited success. Recently, phototherapy, with or without psoralen, was proposed as a successful treatment modality. The aim of this study was to evaluate the effect of broad-band low-dose ultraviolet A (UVA) phototherapy in patients with localized scleroderma, using a new method for evaluation. Twelve patients complaining of morphea were exposed to UVA irradiation at a dose of 20 J/cm2 3 times per week for 20 sessions. Selected covered plaques served as internal controls. The efficacy of therapy was judged clinically by sequential inspection and palpation. In biopsy specimens from exposed and covered plaques stained with hematoxylin and eosin (H & E) and Masson trichrome stains, the concentration of collagen per dermal surface area was measured with the use of a computerized image analyzer. All patients reported remarkable softening of skin lesions, confirmed by sequential palpatory assessment. A significant reduction in the mean concentration of collagen per surface area was detected in the plaques exposed to UVA (the P value being 0.007, P<0.01), whereas in the covered plaques the difference was not statistically significant (the P value being 0.10, P>0.05). The conclusion is that low-dose broad-band UVA phototherapy is a very effective and safe treatment modality for localized scleroderma. PMID- 10823311 TI - Systemic vitamin C and vitamin E do not prevent photoprovocation test reactions in polymorphous light eruption. AB - The possible influence of oxidative stress is discussed in the pathogenesis of polymorphous light eruption (PLE). A double-blind, placebo-controlled study of prophylactic treatment with systemic administration of vitamin C (3 g/d) and E (1500 IU/d) for 8 days was undertaken in 9 patients with PLE (verum, n=4; placebo, n=5). Evaluation of the maximal effects after photoprovocation before and after intake of the antioxidants revealed a reduction of most skin reactions (overall skin reaction, papules/vesicles) in both groups with marked differences in the placebo group. The antioxidants in the doses given and over the time period used did not influence the development of PLE, but might interfere with immunosuppressive effects of repeated photoprovocation tests. PMID- 10823312 TI - Sunscreen application technique in photosensitive patients: a quantitative assessment of the effect of education. AB - Photosensitive patients are known to apply insufficient sunscreen and to neglect several prominently exposed skin sites. The aim of this study was to use fluorescence spectroscopy to assess the effect of education on sunscreen application technique in the short and longer term. Six patients with longstanding photosensitivity conditions were asked to apply an intrinsically fluorescent sunscreen to exposed skin, as they normally would on a sunny day. Detailed fluorescence measurements were taken from 70 sites on the head, neck and arms. Using the previously established dose-response relationship for cream fluorescence, measurements were converted to equivalent thicknesses of cream. Patients were told the results of their sunscreen application assessment and deficiencies in technique were highlighted. Following education, application technique was reassessed twice, after intervals of 2 weeks and 6 months. Before education, sunscreen application was poor with inadequate amounts applied, and prominently exposed sites including ears, temples and neck often missed. At 2 weeks following education, improved application was seen at all sites, and the improvement was sustained at 6 months. Overall, education improved sunscreen application from a baseline median sunscreen thickness of 0.11 mg/cm2 to 0.82 mg/cm2 at 2 weeks and 1.13 mg/cm2 at 6 months (P<0.0001). Notably, median sunscreen thickness on the face improved from a baseline of 0.33 mg/cm2 to 1.51 mg/cm2 at 6 months. These findings demonstrate the importance of careful instruction to patients concerning sunscreen application technique; failure to do this may result in overconfidence in the ability of a sunscreen to protect. The next step is to assess a larger number of photosensitive patients with different diagnoses and to see whether improved technique correlates with improvement in clinical features. PMID- 10823313 TI - The wrist is a reliable body site for personal dosimetry of ultraviolet radiation. AB - The purpose of this study was to investigate the reliability of measuring solar ultraviolet radiation (UVR) doses with personal UV dosimeters worn on the wrist. Individual solar UVR exposure was measured over one day under standardised conditions (One-day Beach Study), and over an extended period of time with varying UV exposure and activities (Holiday Study). Dosimeters of a UV-sensitive spore-film filter type (VioSpor) were placed on the right wrist and on the top of head of the test subjects. The wrist was chosen as being a practical position for personal dosimetry and the head position as an internal control for maximal personal UV doses. The One-day Beach Study took place in the vicinity of Copenhagen in June 1998 over 5 h and included 11 subjects. The Holiday Study included 9 subjects during a period with a mean of 14 days in Scandinavia and Europe from June to September 1998. The head position received the highest UV dose in all subjects in both studies. In both studies, despite considerable individual variation, the mean wrist dose was the same (50%) of that received on the head, although the wrist dose correlated significantly with head dose (P<0.01) only in the Holiday Study. We conclude that the wrist position is a practical and convenient body site for personal dosimetry, yielding reliable results in group exposure studies. PMID- 10823314 TI - Photoscratch testing in systemic drug-induced photosensitivity. AB - Because of numerous false-negative results, photopatch testing is seldom relevant in systemic drug-induced photosensitivity. These false-negative photopatch test results can be attributed to the inability of the drug to penetrate into the epidermis. In order to enhance the penetration of the tested drug into the epidermis, some authors proposed to breach the cutaneous barrier. We performed a prospective study comparing photopatch and photoscratch testing. Fifteen patients presenting with a systemic drug-induced photosensitivity, proved by a favourable outcome after discontinuing the drug, were tested. For each drug, photopatch and photoscratch tests were performed. Two-thirds of the patients had negative photopatch and photoscratch tests with the suspected drugs. Photopatch and photoscratch tests were positive and relevant, respectively, in 3 and 4 patients. Photoscratch tests induced more false-positive results due to irritation confirmed on control subjects. Our study proves that photoscratch tests do not change the sensitivity of phototesting. PMID- 10823315 TI - Timecourse study of UVB-induced cytokine induction in whole mouse skin. AB - Ever since the skin was recognized as a site of immunologic activity, a number of laboratories have studied the production of cytokines by skin cells and the effects of chemicals, allergens, contact irritants and UVB radiation on their production. Most research to date has been carried out using either purified populations of primary cells, or established cell lines. Cytokines, however, do not function in isolation but they appear in human tissues within the context of other cytokines that can, in turn, strongly influence the final biological outcome. Therefore, in vivo studies using whole skin are more physiologically relevant since all cell types are present and interactions among them are allowed to proceed. We report here the results of a detailed timecourse study using whole mouse skin, consisting of both dermis and epidermis, irradiated with either a low or high dose of UVB and analyzed using a Multi-probe RNase protection assay system. The results show that in whole skin the kinetics of cytokine induction are different than what was previously observed in tissue culture. In addition to already known skin-associated cytokines, we report here the presence and UVB induction of cytokines not previously reported. PMID- 10823316 TI - The effect of ultraviolet-B exposure scheduling on the photodamage of hairless mouse skin. AB - In a mouse model, we investigated whether different exposure protocol of ultraviolet-B with the same total doses could induce a different degree of photodamage in mouse skin. Two different exposure frequencies, three times or six times a week, were applied under the condition of weekly same cumulative irradiation dose equally for 10 weeks. Then the photodamage parameters such as skin wrinkling, histochemical dermal change and epidermal and dermal thickness were evaluated. Wrinkle grade, histological assessment by score, and dermal thickness did not reveal any difference between the two groups. However, at irradiation week 10 epidermal thickness of the three times a week irradiation group was significantly thicker than that of the six times a week irradiation group. The same cumulative dose resulted in different epidermal thickness. Our results suggested that exposure frequency or scheduling could influence the epidermal damage by ultraviolet radiation even though the cumulative dose is equal. PMID- 10823318 TI - What's new in photodermatoses PMID- 10823317 TI - The neuro-immuno-cutaneous system and ultraviolet radiation. AB - Numerous cells are closely associated with cutaneous nerve fibers, which through the action of neuropeptides are able to modulate cellular function. Anatomical and physiological links between immune cells and nerves and other cells in skin are so close that we propose the concept of a neuro-immuno-cutaneous system (NICS). Under the impact of ultraviolet radiation (UVR) virtually all properties of the NICS are modified. UVR induces melanin synthesis and immunosuppression, events in which neuropeptides, especially melanocyte stimulating hormone (MSH) and the calcitonin gene-related peptide CGRP, play a role. PMID- 10823319 TI - PUVA improvements will come from molecular biology. PMID- 10823320 TI - Diagnostic impact: a challenge for quantitative nuclear medicine. PMID- 10823321 TI - Principles and technology of PET scanning. PMID- 10823322 TI - PET in oncology I--lung, breast, soft tissue sarcoma. PMID- 10823323 TI - PET in oncology II--other tumours. PMID- 10823324 TI - Clinical PET in cardiology and cardiac surgery. PMID- 10823325 TI - Clinical uses of PET in neurology. PMID- 10823326 TI - Apoptosis: the importance of nuclear medicine. AB - Apoptosis is a genetically controlled, energy-dependent process which removes unwanted cells from the body. Because of its orderly progression, apoptosis is also known as programmed cell death or cell suicide. Once initiated, apoptosis is characterized by a series of biochemical and morphological changes involving the cytoplasm, nucleus and cell membrane. Cytoplasmic changes include cytoskeletal disruption, cytoplasmic shrinkage and condensation; prominent changes in the nucleus include peripheral chromatin clumping and inter-nucleosomal DNA cleavage (DNA ladder formation); and membrane changes include the expression of phosphatidylserine on the outer surface of the cell membrane and blebbing (resulting in the formation of cell membrane-bound vesicles or apoptotic bodies). These events allow the cell to digest and package itself into membrane-bound packets containing autodigested cytoplasm and DNA, which can then be easily absorbed by adjacent cells or phagocytes. An endogenous human protein, annexin V (molecular weight approximately 35,000), has an affinity of about 10(-9) M for phosphatidylserine exposed on the surface of apoptotic cells. Annexin V can be labelled with radionuclides such as iodine or technetium, or positron emitting agents. Experimental studies in cells confirm that fluorescence and 99Tc(m) labelled annexin have comparable affinity for apoptotic cells. In vivo studies with 99Tc(m)-labelled annexin confirm that radiolabelled annexin V can be used to image apoptotic cells/tissues in vivo. In this article, we review experimental data using annexin V imaging and discuss its possible future use to identify apoptosis in vivo. PMID- 10823327 TI - Evaluation of metastatic bone disease with pentavalent 99Tc(m)-dimercaptosuccinic acid: a comparison with whole-body scanning and 4/24 hour quantitation of vertebral lesions. AB - The aim of this study was to establish the value of 99Tcm(V)-DMSA scintigraphy in the detection of metastatic bone lesions and compare the results to 99Tcm-MDP bone scintigraphy. Thirty-four patients presenting with metastatic bone disease (Group 1) and 12 controls with degenerative skeletal lesions (Group 2) were studied. Conventional bone scanning and 99Tcm(V)-DMSA whole-body scanning were performed on all patients. All scans were interpreted visually. Furthermore, lesion-to-normal bone ratios (L/N) in vertebral metastases on the 4 and 24 h bone scans were obtained in 58 lesions of cancer patients and in 23 benign (degenerative) vertebral lesions of the control group. 99Tcm-MDP L/N ratios at 24 h (3.08 +/- 0.32) were significantly higher than those at 4 h (2.48 +/- 0.24) in the malignant foci (P < 0.001). No significant difference was observed in benign lesions (P > 0.05). In 167 (164 metastatic, 3 traumatic) of 186 99Tcm-MDP positive lesions (90%) of Group 1, 99Tcm(V)-DMSA uptake was observed. The remaining 19 lesions (10%) were 99Tcm(V)-DMSA negative. Fourteen of these 19 sites were diagnosed as benign. The remaining five foci were malignant. In four lung cancer metastases showing no 99Tcm-MDP uptake, 99Tcm(V)-DMSA uptake was observed. There was no 99Tcm(V)-DMSA accumulation in any of the 99Tcm-MDP positive degenerative lesions of Group 2. All quantitatively evaluated (n = 42) vertebral metastatic foci and two compression fractures in Group 1 showed 99Tcm(V)-DMSA accumulation and an increased 99Tcm-MDP L/N ratio at 24 h. A total of 36 degenerative lesions (Groups 1 and 2) and one compression fracture (Group 1) showed neither 99Tcm(V)-DMSA uptake nor an increased 99Tcm-MDP L/N ratio at 24 h. Our results indicate that quantitative 4/24 h analysis of vertebral lesions on 99Tcm-MDP scans has a similar diagnostic value to 99Tcm(V)-DMSA imaging in the detection of bone metastases. However, the accumulation of 99Tcm(V)-DMSA in four lung cancer metastases showing no 99Tcm-MDP uptake is encouraging and justifies further research in patients with proven bone metastases and negative bone scans. PMID- 10823328 TI - Quantitative 201Tl SPET imaging in the follow-up of treatment for brain tumour: a sensitive tool for the early identification of response to chemotherapy? AB - The aim of this study was to establish if repeated quantitative 201Tl SPET scanning during follow-up of astrocytoma therapy can provide information that is relevant for clinical management. Sixteen consecutive patients, with histopathologically verified highly malignant astrocytoma, were followed during PCV chemotherapy. Imaging with 201Tl SPET and CT was performed repeatedly over 8 16 weeks until treatment discontinuation, with a maximum follow-up of 74 weeks. Tumour uptake volume (TUV), a measure of metabolically active tumour tissue, was calculated from the SPET images. The reliability of early identification of treatment failure, defined as > 25% tumour volume increase, following one course (week 8) and three courses (week 24) of chemotherapy, was calculated for the two imaging methods. 201Tl SPET positive patients (> 25% tumour volume increase) were compared with 201Tl SPET negative patients in terms of time to treatment discontinuation (TTD) and survival time (ST). The patients were followed with a total of 59 SPET examinations, and treatment was continued for a median 27 weeks (range 16-78 weeks). The comparative reliability of SPET and CT showed the highest sensitivity and accuracy for SPET in the early identification of astrocytoma treatment failure at the week 24 assessment. Patients with positive 201Tl SPET after three courses of chemotherapy had a significantly reduced TTD (P = 0.040) but not significantly reduced ST. Of the ten patients who received concomitant radiation and chemotherapy, five had a small (0-10 ml) TUV at the week 24 assessment. Patients with a TUV > 10 ml at this assessment had a shorter TTD (P = 0.016) and a reduced ST (P = 0.024) compared to patients with a TUV < 10 ml. In conclusion, the assessment of progressive disease by quantitative 201Tl SPET appears to provide information on treatment response, earlier and with a higher reliability than CT. Repeated 201Tl SPET scanning during follow-up of astrocytoma treatment is an alternative tool for the early identification of treatment failure. PMID- 10823329 TI - A non-invasive isotope dilution technique for quantifying hepatic blood flow using radiolabelled red blood cells. AB - Clinically significant changes in hepatic haemodynamics accompany the development of portal hypertension, hepatocellular carcinoma, liver metastases and liver cirrhoses, and after major liver resection. Hepatic blood flow parameters, such as hepatic arterial flow (HAF), hepatic portal flow (HPF), total hepatic blood flow (THBF) and hepatic perfusion index (HPI), are useful adjuncts to the diagnosis of liver pathology, the evaluation of disease progress and prognostication. Here, we describe a non-invasive method that combines the measurement of these parameters in a single study in real time. Red blood cells from eight pigs were labelled with 99Tc(m) using an in-vitro method and re injected into the pigs. Data acquisition over the heart, lungs, liver and kidneys was started immediately and a blood sample was obtained 15 min post-injection. Hepatic arterial flow was determined from the ratio of the maximum gradients between the integrated time-activity curve of the left ventricle and the first pass time-activity curve of the liver before the peak of the kidneys time activity curve. The hepatic perfusion index was determined by comparing the slope of the liver time-activity curve before and after the kidney peak. Hepatic portal flow was determined from the hepatic arterial flow and the hepatic perfusion index, and total hepatic blood flow was determined as the sum of arterial and portal flow. The results were compared against those obtained from a clearance method using 99Tc(m)-DISIDA. The average hepatic perfusion index was 0.38, and the average hepatic arterial flow and hepatic portal flow were 168.3 +/- 52.9 and 274.6 +/- 60.1 ml x min(-1) respectively. The average total hepatic blood flow was 442.8 +/- 53.5 ml x min(-1), while the total hepatic flow determined by 99Tc(m)-DISIDA clearance was 419.7 +/- 62.6 ml x min(-1). No significant difference in total hepatic blood flow was found between the two methods. The results of this study show that it is possible to obtain all hepatic haemodynamics data in a single study using a non-invasive method. PMID- 10823330 TI - Various imaging modalities for the detection of salivary gland lesions: the advantages of 201Tl SPET. AB - The aim of this study was to compare dual-isotope (99Tc(m) and 201Tl) SPET imaging with computed tomography (CT) and magnetic resonance imaging (MRI) in the differentiation of various lesions of the major salivary glands. Twenty-two patients underwent dual-isotope SPET imaging, of whom 12 also had CT and 15 also had an MRI study. The uptake ratio and retention index for 99Tc(m) and 201Tl were calculated by drawing regions of interest on the involved and normal glands. Both CT and MRI were interpreted by two radiologists. All malignant tumours were detected by all three modalities. Warthin's tumours were detected by dual-isotope SPET imaging; however, MRI failed to differentiate Warthin's tumour from pleomorphic adenoma. Of 13 other benign tumours, dual-isotope SPET correctly diagnosed 12. Of 16 tumours showing a cold defect on the 99Tc(m) images, parametric analysis with 201Tl gave an accuracy of 94%, whereas CT gave an accuracy of 70-90%. MRI was 73-91% accurate in differentiating between benign (Warthin's) and malignant tumours. We believe that dual-isotope SPET imaging (99Tc(m) and 201Tl), together with semi-quantitative analysis, is the method of choice for differentiating between various lesions of the major salivary glands. PMID- 10823331 TI - Planar versus SPET imaging in the assessment of condylar growth. AB - The aim of this study was to compare two methods of quantifying 99Tc(m)-methylene diphosphonate uptake in the mandibular condyle. The study groups consisted of 23 patients with mandibular asymmetry and 16 normal volunteers aged 10-30 years. The accuracy and reproducibility of SPET using condyle-to-clivus ratios was compared with planar analysis using condyle-to-L4 (fourth lumbar vertebra) ratios. Quantitative analysis was correlated with semi-quantitative grading by three observers. Normal ranges for condyle-to-L4 and condyle-to-clivus ratios in individuals aged 11 years or over were determined. These ratios are useful in the serial monitoring of patients with condylar hyperplasia to establish when condylar growth has ceased and hence the type of surgery performed. Visual interpretation of condylar activity should use a combination of planar and SPET images and be performed in conjunction with quantitative analysis. Semi quantitative grading on SPET images detected more subtle differences in condylar activity than planar images (using quantitative analysis as a standard). PMID- 10823332 TI - Effect of long-term atrial-demand ventricular pacing on cardiac sympathetic activity. AB - It has been shown that either dual-chamber or atrial pacing may be better than ventricular single-chamber pacing, but the long-term effect of dual-chamber pacing on cardiac sympathetic activity is unclear. The aim of this study was to assess the effect of long-term dual-chamber pacing on cardiac sympathetic activity, compared with atrial pacing and unpaced individuals. We studied 11 patients with dual-chamber pacemakers (Group D), nine with atrial single-chamber pacemakers (Group A) over the long term (mean 44 +/- 36 months) and 10 normal individuals without cardiac pacing. All underwent myocardial 123I metaiodobenzylguanidine (MIBG) imaging to assess cardiac sympathetic activity. The heart-to-mediastinum (H/M) MIBG uptake ratio and the MIBG washout rate from the myocardium were calculated. Echocardiography was performed in all patients with cardiac pacing to assess left ventricular function. In Group D, the H/M ratio on delayed images was significantly lower than that of Group A (1.82 +/- 0.51 vs 2.56 +/- 0.50, P < 0.001) and normal individuals (2.65 +/- 0.35, P < 0.05). The myocardial MIBG washout rate of Group D was significantly higher than that of either Group A (52 +/- 13% vs 36 +/- 8%, P < 0.01) or normal individuals (31 +/- 7%, P < 0.05). Neither the H/M ratio nor MIBG washout rate differed significantly between patients in Group A and normal individuals. Furthermore, the echocardiographic parameters did not differ significantly between the two pacing groups. We conclude that long-term ventricular pacing, even in the presence of atrioventricular synchrony, accelerates cardiac sympathetic activity without deteriorating left ventricular function. PMID- 10823333 TI - Comparison of lung perfusion scintigraphic findings in pulmonary thromboembolism in systemic lupus erythematosus, SLE plus antiphospholipid syndrome, and primary antiphospholipid syndrome. AB - In this study, we compared and reviewed the findings on lung perfusion scans performed in patients with systemic lupus erythematosus (SLE), systemic lupus erythematosus with associated antiphospholipid syndrome (SLE + APS), and primary antiphospholipid syndrome (PAPS), to evaluate the prevalence of pulmonary embolism in restricted samples of the patient groups. Lung perfusion scintigraphy with 99Tc(m)-macroaggregated albumin was performed in 31 patients (SLE = 7; SLE + APS = 14; PAPS = 10). The seven patients with SLE alone and the 10 patients with PAPS had normal perfusion lung scans. Six of the 14 SLE + APS patients showed a segmental uptake defect on multi-view perfusion scans. Thus, the SLE + APS patients were found to have a higher risk of pulmonary thromboembolism than the SLE alone and primary APS patients (P<0.05). The results of our study suggest that lung perfusion scintigraphy should be performed routinely in these patients, even in the absence of pulmonary clinical manifestations, to obtain baseline data for disease outcome and management. PMID- 10823334 TI - Information on nuclear medicine tests on the Internet. PMID- 10823335 TI - Vasoconstrictor reserve in neurally mediated syncope. PMID- 10823336 TI - Forearm vascular responses during orthostatic stress in control subjects and patients with posturally related syncope. AB - The aim of this study was to compare the changes in forearm vascular resistance that occurred during orthostatic stress in asymptomatic volunteer subjects with those in patients with posturally related syncope. The authors hoped firstly that it would indicate the importance of vasoconstriction in the maintenance of blood pressure, and secondly that it might have diagnostic value if there were differences between symptomatic patients and asymptomatic volunteers. Twelve volunteers and 67 patients with unexplained syncope were classified as early or late fainters, based on their endurance of a test of combined head-up tilting and lower-body suction. Responses of vascular resistance were assessed from the ratio of arterial blood pressure (Finapres) to brachial artery blood velocity (Doppler). Changes in vascular resistance were greater in volunteers at all stages of the procedure than in patients. There was, however, no significant difference between the responses of early and late-fainting volunteers. These results demonstrate the importance of vasoconstriction in the resistance to posturally related syncope, and they indicate that assessments of responses of vascular resistance may improve the accuracy of the diagnosis. PMID- 10823337 TI - A new sign of sympathetic neurocirculatory failure: premature ventricular contraction as a "one-beat Valsalva maneuver". AB - Sympathetic neurocirculatory failure (SNF) features orthostatic hypotension and abnormal beat-to-beat blood pressure (BP) responses to the Valsalva maneuver. This article describes a new sign of SNF, based on changes in BP after premature ventricular contractions (PVCs). Records of supine beat-to-beat BP at rest were reviewed from 22 patients with SNF and 52 control subjects. Records were also taken during intravenous infusion of the ganglion blocker trimethaphan from 38 control subjects. In all 10 control subjects who had PVCs, after the postextrasystolic beat, mean arterial pressure increased progressively to levels higher than before the PVC, peaking at about the eighth beat. In contrast, in all 13 patients with SNF who had PVCs, after the postextrasystolic beat, BP decreased to less than baseline and then increased gradually back to baseline. In all 3 control subjects who had at least one PVC before trimethaphan infusion and another during trimethaphan infusion, the post-PVC pressure pattern during trimethaphan infusion resembled that in SNF patients. Because of a brief increase in sympathetic cardiovascular outflows, after a PVC, BP increases progressively to greater than pre-PVC values, mimicking the pressure overshoot after release of the Valsalva maneuver. Just as the absence of the overshoot of BP after release of the Valsalva maneuver supports a diagnosis of SNF, so does the absence of the overshoot after a PVC. PMID- 10823338 TI - Baroreceptor denervation presenting as part of a vagal mononeuropathy. AB - A 48-year-old woman presented with a history of progressive cough, dysphonia, dysphagia, and postural symptoms. Subsequent neurological investigations were consistent with a bilateral vagal mononeuropathy, and neurosarcoidosis was diagnosed after scalene node biopsy. Autonomic investigations including microneurography, neurohormones, and heart rate variability demonstrated arterial and cardiopulmonary baroreflex failure. In addition, parasympathetic control of heart rate was absent and consistent with a bilateral, nonselective lesion in the proximal vagus. PMID- 10823339 TI - Alternating monomeric paresis with decreased skin temperature and hyperhidrosis in a case of thoracolumbar myelopathy. AB - The patient, a Japanese girl with a history suggestive of diffuse encephalitis or acute disseminated encephalomyelitis, developed weakness of the right lower limb accompanied by excessive sweating and decreased skin temperature. Magnetic resonance imaging of the thoracolumbar cord showed abnormal signal intensity with syrinx formation mainly at T12 to L1 vertebral level. Paresis and excessive sweating subsided within 3 to 4 months, but recovery of vasomotor function was delayed. Several weeks later, weakness and skin temperature reduction reappeared on the left side without hyperhidrosis, but responded well to oral prednisolone. The patient showed no recurrence during the subsequent 7 years, and the intramedullary lesion could not be seen with repeated spinal magnetic resonance imaging. PMID- 10823340 TI - Role of dorsal raphe nucleus serotonin 5-HT1A receptor in the regulation of REM sleep. AB - Cholinergic neurons in the laterodorsal (LDT) and the pedunculopontine (PPT) tegmental nuclei act to promote REM sleep (REMS). The predominantly glutamatergic neurons of the REMS-induction region of the medial pontine reticular formation are in turn activated by cholinergic cells, which results in the occurrence of tonic and phasic components of REMS. All these neurons are inhibited by serotonergic (5-HT), noradrenergic, and presumably histaminergic (H2 receptor) and dopaminergic (D2 and D3 receptor) cells. 5-Hydroxytryptamine-containing neurons in the dorsal raphe nucleus (DRN) virtually cease firing when an animal starts REMS, consequently decreasing the release of 5-HT during this state. The activation of GABA(A) receptors is apparently responsible for this phenomenon. Systemic administration of the selective 5-HT1A receptor agonist 8-OHDPAT induces dose-dependent effects; i.e. low doses increase slow wave sleep and reduce waking, whereas large doses increase waking and reduce slow wave sleep and REM sleep. Direct injection of 8-OHDPAT or flesinoxan, another 5-HT1A agonist into the DRN, or microdialysis perfusion of 8-OHDPAT into the DRN significantly increases REMS. On the other hand, infusion of 8-OHDPAT into the LDT selectively inhibits REMS, as does direct administration into the DRN of the 5-HT1A receptor antagonists pindolol or WAY 100635. Thus, presently available evidence indicates that selective activation of the somatodendritic 5-HT1A receptor in the DRN induces an increase of REMS. On the other hand, activation of the postsynaptic 5 HT1A receptor at the level of the PPT/LDT nuclei decreases REMS occurrence. PMID- 10823341 TI - 3,5,3'-Triiodothyronine administered to rat dams during lactation increases milk yield and triglyceride concentration and hastens pups growth. AB - It is known that lactation induces a mild hypothyroid state in rats and other mammals while thyroid hormone administration increases milk secretion in ruminants. The aim of this study was to investigate the effects of a moderate dose of 3,5,3'-triiodothyronine (T3), administered to rat dams during lactation on pups' growth and milk yield and composition. Primiparous Wistar rats with litters adjusted to 10 pups per dam received either tap water or T3 (75 microg/kg x day) in their drinking water from parturition till weaning. Food and water intake of dams and body weight of dams and pups were measured daily. In other groups of rats with similar treatments, milk yield of dams, macronutrient milk composition, and mammary arteriovenous differences for triglycerides (TG) and glucose were also determined. Dams treated with T3 ingested more food and their pups gained more weight than controls. Milk yield, milk TG concentration and glucose extraction by mammary glands were also higher in T3 treated dams. The results show that compensation of the mild hypothyroidism of the lactating rat may contribute to an increase in milk production and lipid levels, leading to an increase in growth of pups. PMID- 10823342 TI - Molecular basis of voltage-dependent delayed rectifier K+ channels in smooth muscle cells from rat tail artery. AB - The molecular basis of voltage-dependent K+ (Kv) current in smooth muscle cells (SMCs) from rat tail artery was investigated by screening transcriptional expression of 15 Kv channel alpha-subunits and 3 Kv beta-subunits using RT-PCR technique. Among Kv genes that encode delayed rectifier Kv currents, mRNAs of Kv1.2, Kv1.3, Kv1.5, Kv2.1, Kv2.2, and Kv3.2 were expressed, but those of Kv1.1, Kv1.6, and Kv3.1 were not detected. The transient outward Kv current-encoding genes Kv1.4, Kv3.3, Kv3.4, Kv4.1- Kv4.3 as well as Kvbeta1, Kvbeta2, and Kvbeta3 were also expressed at mRNA level. Western blot study demonstrated the presence of Kv1.2, Kv1.3, Kv1.5, and Kv2.1, but not Kv3.2 proteins, in tail artery tissue. Immunocytochemistry study confirmed the presence of Kv1.2, Kv1.3, Kv1.5, and Kv2.1 channel proteins in primary cultured single SMCs. Our results represent the first systematic characterization of Kv gene expression in rat tail artery SMCs. PMID- 10823343 TI - Enhancement in extracellular serotonin levels by 5-hydroxytryptophan loading after administration of WAY 100635 and fluoxetine. AB - It has been demonstrated that synthesis of serotonin (5-HT) is dependent on the availability of precursor, as well as the activity of 5-HT neurons. In the present series of experiments, we examined the effects of precursor (5-HTP) loading on extracellular hypothalamic 5-HT after administration of fluoxetine alone or in combination with WAY 100635, a selective 5-HT1A antagonist. In the first experiment, fluoxetine alone (10 mg/kg i.p.) caused 5-HT levels to significantly increase to 150% of basal levels. Subsequent administration of 5 HTP at 10, 20, and 40 mg/kg i.p. caused 5-HT levels to further increase to a maximum value of 254%, 405%, and 618%, respectively. In the second experiment, either vehicle or WAY 100635 (1 mg/kg/hour s.c.) was infused, then fluoxetine (10 mg/kg i.p.) and 5-HTP (10 mg/kg i.p.) were administered. By itself, WAY 100635 led to a slight but significant increase in hypothalamic 5-HT levels one hour after the start of administration (130% of basal levels). In the WAY 100635 treated group, fluoxetine caused an increase to 240% of basal levels after one hour, which rose to 290% of basal levels after two hours. Subsequent administration of 5-HTP further increased 5-HT levels to 580% of basal levels after one hour. In the vehicle-treated group, fluoxetine caused an increase of 160% of basal levels which was stable over two hours, and subsequent administration of 5-HTP led to a slight increase in 5-HT levels of 220% after one hour. These results suggest that combining blockade of 5-HT1A autoreceptors with 5-HT uptake inhibition results in a synergistic increase in synthesis and release of 5-HT when precursor is administered. PMID- 10823344 TI - Dietary troglitazone decreases oxidative stress in early stage type II diabetic rats. AB - Oxidative stress is involved in the initiation and development of atherosclerosis in diabetes. We tested the hypothesis that oxidative stress is already increased in early stage type II diabetes, and that troglitazone may prevent the increase. Three groups of 20 week old rats were studied: untreated Otsuka Long-Evans Tokushima Fatty (OLETF) rats, as an animal model of type II diabetes, OLETF rats treated with troglitazone, and control Long-Evans Tokushima Otsuka (LETO) rats. Plasma lipid hydroperoxides (LOOH) concentration, as an indication of lipid peroxidation, and superoxide dismutase (SOD) activity in the thoracic aorta were measured. Plasma LOOH concentration was significantly higher in non-treated OLETF rats compared to LETO rats and treatment with troglitazone completely prevented this increase. SOD activity was significantly decreased in non-treated OLETF rats compared to LETO rats and troglitazone attenuated the diminution of it. These observations demonstrate oxidative stress is already increased in the early stage of type II diabetes and we confirmed troglitazone has the effect of an antioxidant in vivo. PMID- 10823345 TI - Allopurinol induces renal toxicity by impairing pyrimidine metabolism in mice. AB - We investigated the relationship between the toxic effect of allopurinol and pyrimidine metabolism in mice. Allopurinol-induced increases in plasma transaminase levels in dinitrofluorobenzene (DNFB)-sensitized mice were not affected by uridine. In contrast, plasma creatinine and BUN tended to decrease 18 hr after the last injection of uridine. Both plasma and urinary orotidine (OD) were detected in DNFB-sensitized mice after administration of a single dose of allopurinol. In contrast, TEI-6720, a newly synthesized xanthine oxidase/xanthine dehydrogenase inhibitor, caused neither pyrimidine metabolism abnormality nor renal impairment in DNFB-sensitized mice. Also, normal mice administered high doses of allopurinol showed abnormal pyrimidine metabolism together with renal toxicity which could be ameliorated by uridine, indicating that allopurinol essentially causes pyrimidine metabolism abnormality leading to renal impairment. In DNFB-sensitized mice, allopurinol increased urinary OD excretion to an extent similar to that in normal mice administered the same dose of allopurinol. However, renal impairment by allopurinol was more striking in DNFB-sensitized mice than in normal mice. Histopathological observations showed that allopurinol induced calculus formation in the collecting tubules and papillary duct. Calculus formation was increased by DNFB and decreased by uridine. These observations indicate that the enhancement of the renal toxicity of allopurinol by DNFB sensitization may be due to some biological interactions between DNFB and allopurinol. In humans, it is possible that there are some biological interactions which serve to enhance the toxicity of allopurinol, resulting in the development of allopurinol hypersensitivity syndrome (AHS). In contrast, TEI 6720, had no effect on pyrimidine metabolism and showed no toxic effect. PMID- 10823346 TI - Influence of experimental diabetes on regulatory mechanisms of vascular response of rabbit carotid artery to acetylcholine. AB - The purpose of this study was to analyse the influence of experimental diabetes on vascular response of rabbit carotid artery to acetylcholine (Ach). We compared the Ach-induced relaxant response of isolated arterial segments obtained from both control and diabetic animals. To assess the influence of the endothelium, this cell layer was mechanically removed in some of the arterial segments ("rubbed arteries") from each experimental group. Ach induced a concentration related endothelium-mediated relaxation of carotid artery from control rabbits that was significantly higher with respect to that obtained in diabetic animals. Pre-treatment with N(G)-nitro-L-arginine (L-NA) induced a concentration-dependent inhibition of relaxant response to Ach, which was significantly higher in carotid arteries isolated from diabetic rabbits. Incubation of rubbed arteries with L-NA almost abolished the relaxant response to Ach in arterial segments from both control and diabetic animals. Indomethacin potentiated Ach-induced response of carotid arteries from control rabbits, without modifying that obtained in those from diabetic animals. Aminoguanidine did not significantly inhibit the relaxant action of Ach in arterial segments from either control or diabetic rabbits. These results suggest that diabetes impairs endothelial modulatory mechanisms of vascular response of rabbit carotid artery to Ach. This endothelial dysfunction is neither related with a lower release of nitric oxide (NO) or prostacyclin. Diabetes impairs the production of some arachidonic acid vasoconstrictor derivative. There has been observed an increased modulatory activity of NO, but this is not related with the expression of an inducible isoform of NO synthase. PMID- 10823347 TI - Studies on the prolactin-releasing mechanism of histones H2A and H2B. AB - In previous studies we demonstrated that histone preparations possess multiple effects in vivo on pituitary hormone secretion. We have now studied the specificity and signal transduction pathways involved in the prolactin (PRL) releasing activity of histones H2A and H2B on perifused and incubated rat pituitary cells. In the perifusion experiments, freshly dispersed pituitary cells were packed into short columns and were continuously perifused with serum-free medium. The substances to be tested (stimuli) were pumped through the perifusion circuit, at the end of which perifusate fractions were collected and PRL measured by specific RIA. In the incubation studies, freshly dispersed pituitary cells were incubated in a metabolic incubator with different stimuli at different doses and for varying times. Perifusion of cells with median eminence extract (1/30), histone H2A (30 microM) or histone H2B (30 microM), generated clear PRL release responses. Cells incubated with histone H2A and H2B showed a dose- and time dependent stimulatory effect on PRL release which, for H2A, was blocked by peptide MB-35, an 86-120 amino acid synthetic fragment of histone H2A. The polycation, poly-lys was unable to mimic the action of histones. To detect the possible signal transduction pathways involved in the response of lactotrophs to histones, cells were incubated with the calcium ionophore A23187, the calcium chelator EGTA, the intracellular phosphoinositide enhancer LiCl, the intracellular cAMP enhancers caffeine, NaF and forskolin, and the protein kinase C inhibitor, trifluoperazine (TFP). Both EGTA (or EGTA plus A23187 ionophore) and TFP were able to reduce significantly the response of lactotrophs to histones. Our results confirm previous evidence that histones may act as hypophysotropic signals. The data also suggest that calcium- and diacylglycerol-associated pathways participate in these effects. PMID- 10823348 TI - In vivo effects of partial phosphorothioated AT1 receptor antisense oligonucleotides in spontaneously hypertensive and normotensive rats. AB - Partial phosphorothioate (PS) antisense oligodeoxynucleotides (ODNs) targeted against rat AT1 receptor mRNA have been used to control blood pressure in normotensive (WKY) and spontaneously hypertensive (SHR) rats. Molecules were injected intracerebroventricularly (i.c.v., right lateral ventricle) in freely moving animals. The antisense ODN lowered the mean arterial pressure (MAP) 24 hours (-43 mmHg+/-10) and 48 hours (-30 mmHg+/-13) after injection, while the control ODN molecule had no significant effects. The observed decrease of blood pressure was due to a specific inhibition of AT1 receptor gene expression, since the level of its mRNA, monitored by reverse transcription (RT)- polymerase chain reaction (PCR), was significantly reduced by antisense molecule (-40%), compared to sense one. In normotensive rats no effect on MAP have been observed, while AT1 receptor gene expression is reduced (-40%) by antisense treatment. It is known that SHRs have an enhanced basal activity of the central renin-angiotensin system that induces an increase in central sympathetic outflow. Instead in WKY rats the central sympathetic outflow is not conditioned by the enhanced activity of brain renin-angiotensin system. Therefore in normotensive rats although partial PS ODN reduces the AT1 mRNA level this will not result in a modification of the sympathetic outflow and no change in MAP level would be observed. PMID- 10823349 TI - Hypoalgesic effect of millimeter waves in mice: dependence on the site of exposure. AB - Based on a hypothesis of neural system involvement in the initial absorption and further processing of the millimeter electromagnetic waves (MW) signal, we reproduced, quantitatively assessed and compared the analgesic effect of a single MW treatment, exposing areas of skin possessing different innervation densities. The cold water tail flick test (cTFT) was used to assess experimental pain in mice. Three areas of exposure were used: the nose, the glabrous skin of the right footpad, and the hairy skin of the mid back at the level of T5-T10. The MW exposure characteristics were: frequency = 61.22 GHz; incident power density = 15mW/cm2; and duration = 15 min. The maximum hypoalgesic effect was achieved by exposing to MW the more densely innervated skin areas--the nose and the footpad. The hypoalgesic effect in the cTFT after MW exposure to the murine back, which is less densely innervated, was not statistically significant. These results support the hypothesis of neural system involvement in the systemic response to MW. PMID- 10823350 TI - A novel effect of polymorphonuclear leukocytes in the facilitation of angiogenesis. AB - The purpose of this study was to examine whether the adhesion of polymorphonuclear leukocytes (PMNs) to endothelial cells and/or reactive oxygen species (ROS) released from PMNs are responsible for inducing angiogenesis. Angiogenesis was assessed by tube formation using endothelial cells obtained from bovine thoracic aorta (BAECs) grown on a layer of collagen type I. Addition of PMNs to BAECs weakly induced angiogenesis. The angiogenesis induced by PMNs alone was further enhanced by treatment of the PMNs with N-formyl-methionyl-leucyl phenylalanine (FMLP), a selective activator of PMN. The involvement of PMN adhesion to BAECs via adhesion molecules in angiogenesis was investigated by using monoclonal antibodies against E-selectin and intercellular adhesion molecule-1 (ICAM-1). These antibodies blocked both the PMN adhesion to BAECs and the enhancement of angiogenesis induced by FMLP-treated PMNs. Furthermore, the enhancement of angiogenesis by FMLP-treated PMNs was blocked by catalase, a scavenging enzyme of H2O2, but not by superoxide dismutase (SOD). These results suggest that PMNs induce angiogenesis in vitro, and that the mechanism of stimulation of angiogenesis by PMNs may involve the adherence of PMNs to endothelial cells via E-selectin and ICAM-1, and H2O2, but not superoxide. Thus, activated PMNs in pathological states may not only induce tissue injury, but may also function as regulators of angiogenesis. PMID- 10823351 TI - Activation and inhibition of mast cells degranulation affect their morphometric parameters. AB - Activation of mast cells, the key cells of allergic inflammation, causes typical morphological changes associated with an increase in volume, that is a function of area and perimeter. The purpose of this study was to evaluate the effect of mast cell activation to degranulate, carried out by the secretagogue Compound 48/80, and of inhibition of this activation carried out by Nedocromil sodium, a mast cell stabilizing drug, on mast cell area, perimeter and shape factor by a computerized image analyzer. Mast cells were isolated and purified by peritoneal lavage of rats (purity >98%) and co-cultured with mouse 3T3 fibroblasts to which they adhere. Cultures were incubated for 10 min at 37 degrees C with culture medium alone (Enriched Medium) or Enriched Medium containing either Nedocromil (10(-4) M) or Compound 48/80 (0.3 microg/ml) or Compound 48/80 and Nedocromil (0.3 microg/ml and 10(-4) M respectively). Supernatants were then assessed for histamine release, as a marker of mast cell activation and the cell monolayers were fixed and stained with an alcoholic-acidic toluidine blue solution and examined with a computerized image analyzer connected with a light microscope. Mast cells incubated in Enriched Medium or Nedocromil possessed similar morphometric parameters. Mast cells activated with Compound 48/80 (70% histamine release) had a significant increase in area and perimeter and a decrease in shape factor in comparison to mast cells in Enriched Medium alone. Simultaneous incubation of mast cells with Compound 48/80 and Nedocromil significantly inhibited their histamine release (36% histamine release) and the increase in area and perimeter, but did not affect significantly their shape factor, in comparison with mast cells incubated with Compound 48/80 alone. These data clearly show that there is a relationship between mast cell activation, consequent histamine release and changes in cell area, perimeter and shape factor and that Nedocromil not only inhibits mast cell histamine release but also the activation induced morphometric changes in mast cells. PMID- 10823352 TI - Differential contractile actions of reactive oxygen species on rat aorta: selective activation of ATP receptor by H2O2. AB - This study aims to examine the effects of different reactive oxygen species (ROS) on the resting tension of endothelium-denuded rat aortic rings. In these preparations, H2O2 (30 microM) induced a fast and transient contraction, which could be abolished by pretreatment of catalase (800 U/ml), but not affected by superoxide anion scavenger, superoxide dismutase (SOD; 150 U/ml) or the hydroxyl free radical scavenger, DMSO/mannitol (each 3 mM). In contrast, pyrogallol, a putative superoxide anion donor, induced a biphasic contraction, which could be abolished by SOD, but not by catalase or DMSO/mannitol. Unlike H2O2 and pyrogallol, Vitamin C(VitC)/Fe2+ (each 100 microM), a commonly used hydroxyl radical-generating system, triggered a tonic contraction which could be prevented by DMSO/mannitol, but not by SOD or catalase. Interestingly, H2O2-induced contraction could be concentration-dependently (10-100 microM) inhibited by suramin and reactive blue-2 (RB-2), two widely used ATP receptor antagonists. On the other hand, suramin or RB-2, at concentration up to 100 microM, affected neither pyrogallol nor VitC/Fe2+-induced contraction. In conclusion, we showed for the first time that different ROS could contract rat aorta with different mechanisms of action, and H2O2 elicits a transient contraction probably as a result of the ATP receptor activation. PMID- 10823353 TI - Pollen availability for predaceous mites on apple: spatial and temporal heterogeneity. AB - It has been suggested that an abundance of alternate food early in the spring may be critical to the ability of generalist predaceous mites to suppress spider mite pests. One alternate food that is typically very abundant in spring is wind dispersed pollen. Here we investigate, at several spatial scales, the heterogeneity in the availability of pollen to predaceous mites on apple. We found pollen to be abundant on apple leaves very soon after they opened (> 100 grains/cm2), and that the dominant pollen types at this time were wind-dispersed tree pollens (Betulaceae and Pinaceae). We found that most of the spatial variation in pollen abundance occurred at either small spatial scales (within trees) or very large spatial scales (among orchard blocks). Variability among orchards was clearly influenced by the surrounding vegetation, and probably also by the management regime (frequency of mowing). Spatial heterogeneity in pollen availability may affect the build-up of predatory mite populations in the spring, as we found early season abundances of Typhlodromus pyri (Phytoseiidae) and Zetzellia mali (Stigmaeidae) to be better correlated with early season pollen density than with abundance of mite prey (Aculus schlechtendali). PMID- 10823354 TI - Some effects of pre-release host-plant on the biological control of Panonychus ulmi by the predatory mite Amblyseius fallacis. AB - Amblyseius fallacis Garman has been selected for pyrethroid resistance and mass reared for experimental release as a biological control agent for tetranychid mites on a number of crops in Canada. Several releases of this predator onto apple and peach trees have failed to result in the establishment of A. fallacis, or in the biological control of Panonychus ulmi Koch. Here, we test the hypothesis that the change of host-plant at the time of release is a critical factor in the establishment of A. fallacis for biological control of P. ulmi. Functional and numerical response studies were undertaken on two populations of A. fallacis: a wild strain collected from the canopy foliage of an apple orchard near Vineland, Ontario; and a second strain reared on bean plants in a commercial insectary with Tetranychus urticae as prey. Each population consumed significantly more P. ulmi and produced significantly more eggs when on leaf disks from the plant species they were reared on, than on leaf disks from the novel host plant. A further experiment was conducted to determine if establishment and biological control of mass-reared A. fallacis could be affected by rearing a population for a short term on apple leaves prior to release on apple trees. Three release treatments were made into potted apple trees in a glasshouse, using predators commercially mass-reared on bean and T. urticae: A. fallacis released directly; A. fallacis reared in the laboratory for four weeks on bean and T. urticae; A. fallacis reared on apple leaves and T. urticae for four weeks. They were compared with a control treatment lacking predator release. Contrary to results of the functional and numerical response studies, no difference was observed between release treatments. All release treatments adding A. fallacis resulted in a similar, if limited, degree of biological control of P. ulmi. These results indicated that there may be short-term effects of host plant on the establishment of A. fallacis and biological control of P. ulmi, which in our study were observed as an initial reduction of the predatory response. However, in a test, the predators appeared to overcome these short-term effects and successfully established on the new host-plant to control P. ulmi. PMID- 10823355 TI - First detection in Israel of fluvalinate resistance in the varroa mite using bioassay and biochemical methods. AB - The aim of this study was to explore the extent of varroa mite resistance to fluvalinate in Israel and to determine the underlying biochemical mechanism. Assays at different apiaries indicated varroa mite resistance at three of the five sites tested. Dose response assays conducted with tau-fluvalinate on mites obtained from different sites indicated uneven resistance. A monooxygenase assay revealed an increased rate (approximately 20-fold) of activity in mites that were not controlled by the pesticide, as compared to activity in mites from untreated colonies. A minor, 1.5-2.5 fold, increase of esterase activity was also noted in the resistant mites. This first demonstration of a fluvalinate-resistance mechanism in varroa mites points to the need for more vigorous resistance management practices to control the pest. PMID- 10823356 TI - Pirimiphos-methyl resistance in two stored product mites, Acarus siro and Acarus farris, as detected by impregnated paper bioassay and esterase activity assays. AB - The response to pirimiphos-methyl, in one strain of Acarus farris and two strains of Acarus siro, was assessed using an impregnated filter paper bioassay and by the selection of adults following exposure to pirimiphos-methyl. It was concluded that one of the strains of A. siro was resistant to pirimiphos-methyl and that a major resistance mechanism was involved. The second strain of A. siro gave a response similar to that of a laboratory strain unexposed to organophosphates and was considered to be susceptible. The A. farris strain responded to selection at the ED50 but not at the ED99, and it was concluded that a minor resistance mechanism is present in this strain. Assays of esterase activity were used to attempt to identify the biochemical mechanisms involved in the resistance detected by the bioassays. The A. farris and susceptible A. siro strains showed similar levels of esterase activity but the esterase activity of the resistant A. siro strain was significantly greater. An increase in esterase activity followed selection of both the A. farris strain and the resistant A. siro strain. An acetylcholinesterase assay showed no significant difference between the susceptible and pirimiphos-methyl selected strains of A. siro. The results suggest that esterases are involved in the resistance to pirimiphos-methyl found in A. siro and A. farris but that in A. siro, at least, other mechanisms may also be present. PMID- 10823357 TI - Photographic sampling: a photographic sampling method for mites on plants. AB - A photographic sampling method for mites on plants was evaluated using Tetranychus urticae and Phytoseiulus persimilis on pepper plants. It was found to be 92% accurate for T. urticae eggs and 98% accurate for P. persimilis eggs at densities up to 45 eggs per cm2 for T. urticae, and up to 3 eggs per cm2 for P. persimilis. The motiles of the two species were not confused, nor were they confused with exuviae or other matter. PMID- 10823358 TI - Modus operandi of oviposition in Dermacentor reticulatus (Acari: Ixodidae). AB - The process of oviposition in D. reticulatus was observed and found to be a sequence of exactly coordinated, interlocking events independent of the phase of oviposition. The average period of oviposition in the investigated ticks was 31.6 days at 20 degrees C and 95% relative humidity. The number of eggs deposited on each day increased until reaching a maximum on the fifth day of oviposition and then decreased continuously. As a result, most of the eggs were deposited during the initial phase of oviposition. The total number of eggs was proportional to the ticks' weight replenishment. Egg-laying commenced with the lowering of the capitulum and the simultaneous spread of the pedipalps which were lowered to the body wall embracing the genital aperture on both sides. Immediately afterwards the cuticular sac of Gene's organ was pushed out and retracted several times. At the cuticular sac's maximum extension, the vestibulum vaginae prolapsed, forming the ovipositor as an extended tube which handed over an egg to the two horns of the cuticular sac after a brief, but intensive, contact with the cuticular sac. Then the vestibulum vaginae invaginated, the pedipalps closed, and the cuticular sac was retracted. Finally, the egg was transported onto the dorsal area of the tick by means of a vigorous rising of the capitulum. During the course of oviposition most of the events, especially the period of egg embracement by the cuticular sac, were prolonged, as was the total time for laying one egg. Similarly, the intervals between successive egg-laying processes increased continuously. The number of eggs deposited was not dependent on the functional ability of Gene's organ, as shown by similar numbers of deposited eggs from ticks with and without mechanical blocking of the cuticular sac. But the participation of the organ in the process of oviposition proved to be a prerequisite for the viability of the eggs. Larvae developed and hatched only from those eggs which were deposited from ticks with an undisturbed Gene's organ. In comparison, eggs without contact to the cuticular sac of Gene's organ dried up and shrivelled immediately after being deposited and did not hatch. Consequently, it strongly suggests, together with the results from other studies, that Gene's organ covers the eggs with a secretion that prevents the loss of water. PMID- 10823359 TI - Vector competence of Ixodes angustus (Acari: Ixodidae) for Borrelia burgdorferi sensu stricto. AB - The vector competence of Ixodes angustus for Borrelia burgdorferi sensu stricto (s.s.) was investigated in the laboratory. The larval progeny of female ticks from Washington State were placed on Swiss-Webster mice that had been inoculated intravenously with 10(8) spirochetes each of a Californian isolate of B. burgdorferi. Spirochetes were detected in 6 (12%) of 50 nymphs derived from larvae that had fed on these animals. Ten nymphs from the same cohort of experimentally infected ticks were placed on each of 4 naive deer mice (Peromyscus maniculatus). One of the mice seroconverted to B. burgdorferi and spirochetes were isolated from its ear tissues 4 weeks after exposure to ticks. Further vector competence trials were conducted with I. angustus ticks from California. Larvae were fed on deer mice that had been inoculated intradermally with B. burgdoferi along with larvae of I. spinipalpis as a comparison group. There was no significant difference in the prevalence of infection in nymphs of I. angustus (8.2%) versus those of I. spinipalpis (12.1%). We conclude that I. angustus is a competent experimental vector of B. burgdorferi s.s. and its efficiency for acquiring and transstadially passing such spirochetes is similar to that of I. spinipalpis. PMID- 10823360 TI - Sparfloxacin: a review. AB - BACKGROUND: The continuing increase in the rate of penicillin and cephalosporin resistance among respiratory pathogens and of cross-resistance to macrolide antibiotics has led to the recommendation that fluoroquinolone antibiotics be used to treat high-risk patients with community-acquired pneumonia (CAP) and acute bacterial exacerbations of chronic bronchitis (ABECB). OBJECTIVE: This review focuses on sparfloxacin, an oral fluoroquinolone, discussing its mechanism of action, activity, pharmacokinetic characteristics, safety, and efficacy in CAP and ABECB. METHODS: Studies were identified by a MEDLINE search of the literature from 1990 to 1999, supplemented by educational materials from conferences and symposia. RESULTS: Sparfloxacin is active against the major respiratory pathogens and against the atypical pathogens in pneumonia that are being reported with increasing frequency. Its long half-life permits once-daily dosing. In large trials in CAP and ABECB in which all isolates were susceptible to both comparators, sparfloxacin was found to have similar efficacy to erythromycin, cefaclor, amoxicillin, ofloxacin, and clarithromycin. Its safety profile is similar to that of the macrolides and other quinolone antimicrobial agents. Photosensitivity, nausea, and diarrhea are the most common adverse events reported in clinical trials of sparfloxacin. Its use is contraindicated in patients with QTc-interval prolongation. CONCLUSION: The increasing prevalence of beta-lactam- and macrolide-resistant bacteria in respiratory infections emphasizes the need for newer agents such as the fluoroquinolones. The choice between agents should be based on activity against the relevant respiratory pathogens in high-risk patients. PMID- 10823361 TI - Comparison of brimonidine with latanoprost in the adjunctive treatment of glaucoma. ALPHAGAN/XALATAN Study Group. AB - OBJECTIVE: This study compared brimonidine with latanoprost as adjunctive therapy for the treatment of open-angle glaucoma and ocular hypertension. BACKGROUND: Patients with open-angle glaucoma or ocular hypertension often require >1 medication to achieve control of intraocular pressure (IOP). Both brimonidine and latanoprost effectively lower IOP, but no previously reported clinical trials have directly compared these agents as adjunctive therapy. METHODS: This was a prospective, randomized, investigator-masked, multicenter, parallel-design clinical trial. Forty patients (69 study eyes) with uncontrolled IOP of < or =34 mm Hg while using a topical beta-blocker plus dorzolamide or pilocarpine were randomly assigned to receive either brimonidine 0.2% BID or latanoprost 0.005% QD over 6 months as adjunctive therapy. Tolerability was assessed by reports of adverse events, and efficacy was determined by reduction in IOP from baseline. Clinical success was defined as the achievement of a > or =15% reduction in IOP from baseline. RESULTS: There were no significant between-group differences in any demographic variable. Most patients in each group were white, had open-angle glaucoma, and were being treated with a nonselective beta-blocker and dorzolamide. When brimonidine or latanoprost was used as an adjunctive agent with a beta-blocker and dorzolamide or pilocarpine, the rates of clinical success at month 1 were 85% (17/20 patients) with brimonidine versus 65% (13/20 patients) with latanoprost (P = 0.144). Overall mean IOP reduction at month 1 was 4.60+/ 0.62 mm Hg (22.8%; P < 0.001) with brimonidine and 3.43+/-0.62 mm Hg (17.2%; P < 0.001) with latanoprost, with no significant differences between groups (P = 0.219). Among the patients with an inadequate IOP-lowering response (<15% reduction from baseline), the mean IOP reduction was 0.36+/-0.66 mm Hg with latanoprost (n = 7) and 0.50+/-2.18 mm Hg with brimonidine (n = 3). Brimonidine and latanoprost had comparable IOP-lowering efficacy in patients receiving concomitant pilocarpine therapy (mean change in IOP of -4.23 mm Hg vs -3.75 mm Hg, P = 0.173). In patients concurrently treated with dorzolamide, brimonidine produced a mean change in IOP of -5.29 mm Hg, compared with a mean change of 3.21 mm Hg in the latanoprost group (P = 0.159). Both brimonidine and latanoprost were well tolerated. Few adverse events leading to discontinuation were observed with either drug regimen (n = 2 with brimonidine; n = 0 with latanoprost). CONCLUSIONS: Both brimonidine 0.2% BID and latanoprost 0.005% QD were well tolerated and reduced IOP in most patients when used as third-line adjunctive therapy. However, clinical success was achieved by 17 of 20 patients (85%) who received brimonidine, compared with 13 of 20 patients (65%) who received latanoprost (P = 0.144). These results suggest that brimonidine 0.2% BID may be more reliable than latanoprost 0.005% QD as adjunctive therapy for glaucoma and ocular hypertension. PMID- 10823362 TI - Efficacy and tolerability of meloxicam in an observational, controlled cohort study in patients with rheumatic disease. AB - BACKGROUND: Meloxicam is a nonsteroidal anti-inflammatory drug (NSAID) that inhibits cyclooxygenase-2 without affecting platelet aggregation. OBJECTIVE: The goal of this study was to assess the efficacy and tolerability of meloxicam under natural prescribing conditions for up to 6 months. METHODS: This was a multicenter, prospective, observational cohort study. Participating centers were randomized to 1 of 2 groups: the meloxicam-only group, and the group who received comparator NSAIDs (ie, diclofenac, ibuprofen, piroxicam, or indomethacin). RESULTS: A total of 4526 patients received either meloxicam (n = 2530) or a comparator NSAID (n = 1996). Treatment groups were comparable in terms of observed efficacy measures. Meloxicam patients had significantly lower rates of reported gastrointestinal (GI) adverse drug reactions (1.80% vs 3.20%; P = 0.003), including dyspepsia (0.08% vs 0.35%; P = 0.049), abdominal pain (0.91% vs 1.90%; P = 0.006), gastritis (0.08% vs 0.60%; P = 0.002), and GI bleeding (0.08% vs 0.50%; P = 0.007), compared with those receiving comparator NSAIDs. CONCLUSION: There was evidence that meloxicam was prescribed preferentially to patients who had not responded to NSAIDs previously, who had previously experienced NSAID-induced side effects (and so were at high risk for developing NSAID-induced GI toxicity), or who were more seriously impaired. The nonrandomized and unblinded nature of this study limits the conclusions that can be drawn concerning efficacy or tolerability. Nevertheless, the study results are consistent with the favorable GI tolerability seen with meloxicam in double-blind comparative trials. PMID- 10823363 TI - Comparison of an extract of hypericum (LI 160) and sertraline in the treatment of depression: a double-blind, randomized pilot study. AB - BACKGROUND: Hypericum (St. John's wort) has been shown to be as efficacious and well tolerated as standard antidepressants in the treatment of depression but has not been compared with selective serotonin reuptake inhibitors (SSRIs). OBJECTIVE: This study compared hypericum and the SSRI sertraline in the treatment of depression. METHODS: In a double-blind, randomized study conducted in a community hospital, 30 male and female outpatients (19 women, 11 men; mean age, 45.5 years) with mild to moderate depression received 600 mg/d of a standardized extract of hypericum (LI 160) or 50 mg/d sertraline for I week, followed by hypericum 900 mg/d or sertraline 75 mg/d for 6 weeks. RESULTS: The severity of symptoms, as assessed by scores on the Hamilton Rating Scale for Depression (HAM D) and the Clinical Global Impression scale, was significantly reduced in both treatment groups (P < 0.01). Clinical response (defined as a > or =50% reduction in HAM-D scores) was noted in 47% of patients receiving hypericum and 40% of those receiving sertraline. The difference was not statistically significant. Both agents were well tolerated. A post hoc power analysis indicated that failure to reach statistical significance between treatments resulted primarily from an absence of clinical differences rather than the small sample size. CONCLUSION: The hypericum extract was at least as effective as sertraline in the treatment of mild to moderate depression in a small group of outpatients. PMID- 10823364 TI - Management of patients with Bacilli Calmette-Guerin-refractory carcinoma in situ of the urinary bladder: cost implications of a clinical trial for valrubicin. AB - OBJECTIVE: This study was undertaken to identify the expected first- and second year clinical costs associated with intravesical valrubicin therapy, using a decision analytic model, for patients with Bacilli Calmette-Guerin (BCG) refractory carcinoma in situ (CIS) of the urinary bladder. BACKGROUND: Cancer of the urinary bladder is the fourth most common malignancy in men and the sixth most common noncutaneous carcinoma overall. One histopathologic stage of bladder cancer is CIS, for which BCG intravesical immunotherapy is the first-line therapy. Radical cystectomy has been recommended for patients with CIS who do not respond to or become refractory to therapy with BCG. Surgery, however, may not be appropriate for all patients, especially those who are ineligible for the lengthy procedure because of advanced age or comorbidities and those who prefer alternative nonsurgical management. For these groups, intravesical valrubicin therapy is a plausible alternative. METHODS: Models were developed and populated with data from 1 open-label study of 90 patients, information from the medical literature, and input from clinical experts. The analysis was conducted from the payor perspective for direct costs only. RESULTS: Our data indicate that first- and second-year expected costs for valrubicin therapy are $19,912 and $23,496, respectively. Expected cost for radical cystectomy was also evaluated, since some patients may have no other option if drug therapy fails. CONCLUSION: Our cost consequence analysis and clinical data provide decision-makers with tools to aid in global budgetary projections of fractional and total expected health care costs associated with the management BCG-refractory CIS of the urinary bladder. PMID- 10823365 TI - Savings in the cost of caring for patients with Alzheimer's disease in Canada: an analysis of treatment with rivastigmine. AB - OBJECTIVE: To estimate per-patient potential cost savings using rivastigmine in the treatment of Alzheimer's disease (AD) in Canada. BACKGROUND: In recent years, new members of a class of pharmaceuticals known as cholinesterase inhibitors have been introduced for the treatment of patients with AD. Two recent studies conducted in the United Kingdom and the United States estimated potential cost savings from the new cholinesterase inhibitor rivastigmine. The present study combined the disease-progression model used in those 2 studies with Canadian costs to estimate per-patient potential savings resulting from the treatment of AD in Canada. METHODS: Efficacy data from 2 pivotal, phase III clinical trials of rivastigmine were used in a hazard model of disease progression to estimate long term differences in cognitive functioning between patients receiving rivastigmine and patients receiving no treatment. We used the Mini-Mental State Examination (MMSE) score as our measure of disease progression. We also used Canadian costs of AD care, estimated as a function of MMSE score, to estimate cost savings experienced by treated patients compared with patients receiving no treatment. All costs and cost savings are presented in 1997 Canadian dollars. We used a societal perspective in this analysis. RESULTS: Rivastigmine was estimated to delay the transition to more severe stages of AD by up to 188 days for patients with mild AD after 2 years of treatment. For patients with mild-to-moderate and moderate disease, this delay was estimated to be 106 and 44 days, respectively. For patients with the mild stage of AD, estimated average daily cost savings (excluding the cost of rivastigmine) ranged from Can $0.45 per patient per day at 6 months to Can $6.44 per patient per day after 2 years of treatment. For all patients, these estimated average daily cost savings ranged from a low of Can $0.71 per patient per day after 6 months of treatment to a high of Can $4.93 per patient per day after 2 years. CONCLUSION: On average, treatment with rivastigmine yields savings in the direct cost of caring for AD patients that exceed the cost of the drug after 2 years of treatment. PMID- 10823366 TI - Inhaled corticosteroid use and associated outcomes in elderly patients with moderate to severe chronic pulmonary disease. AB - OBJECTIVE: This study was undertaken to examine the impact of select demographic, clinical, and medication-related factors on elderly patients' medication adherence to inhaled corticosteroid therapy. It also examined the impact of medication adherence on health outcomes and on health care service utilization and costs. BACKGROUND: Asthma in the elderly is a growing concern because of its increasing prevalence and the increase in asthma-related deaths. Poor medication utilization could be contributing to this trend in the elderly population and should be examined. METHODS: This retrospective, longitudinal, 2-year cohort study included 1595 patients aged > or =65 years with moderate to severe chronic pulmonary disease who were enrolled in a health maintenance organization. The study used sequential regression analyses to model (1) medication adherence and (2) health care service utilization and charges as functions of adherence. RESULTS: With other factors controlled for, this study found that clinical and medication-related variables including comorbidities, additional complications, and number of medications were predictive of adherence to prophylactic inhaled corticosteroid therapy in this population. Poorer medication adherence was associated with a 5% increase in total annual physician visits (P < 0.05). Better medication adherence was associated with a 20% decrease in annual hospitalization (P < 0.05). Additional pulmonary complications and severe comorbidities were associated with increases in health care utilization and costs. Patients with better adherence to prophylactic therapy were few, and the beneficial effects of better adherence were not significant when the population averages were considered. CONCLUSIONS: The results of this study indicated that the beneficial effects of better adherence to prophylactic inhaled corticosteroid therapy tended to diminish in a population with relatively poor medication adherence. Findings of this study support a policy of selectively targeting elderly patients at risk for morbidity and implementing monitoring, education, and management programs to increase medication-adherence behavior. This policy could lead to optimal resource utilization and management of chronic pulmonary disease. PMID- 10823367 TI - Analytic dimensions of a prescription-medication benefit in medicare. AB - BACKGROUND: Many analysts believe that the lack of coverage for outpatient prescription medications represents a conspicuous deficiency in the Medicare benefits package. OBJECTIVE: This paper uses insurance theory to design and estimate the costs of a Medicare catastrophic-medication outpatient benefit. RESULTS: For efficiency and equity purposes, and to accommodate the tradeoff between the cost to the federal government and the insurance value of such a benefit to Medicare enrollees, we favor a benefit that would be means-tested by employing deductibles, coinsurance rates, and catastrophic limits, all of which would be progressively graduated for 7 household income classes. For equity reasons, we propose that the government's share of the medication benefit be financed from the general tax fund, using the progressive income tax. Another source of potential savings within the Medicare program that could pay for a medication benefit would be elimination of fraud, waste, and abuse. CONCLUSIONS: Because our proposal addresses both the efficiency and equity dimensions of a Medicare outpatient medication benefit, we believe it is worthy of serious consideration by both policymakers and Congress. PMID- 10823368 TI - A comparative economic analysis of simvastatin versus atorvastatin: results of the Surrogate Marker Cost-Efficacy (SMaC) study. PMID- 10823369 TI - Advances in therapeutic drug monitoring for immunosuppressants: a review of sirolimus. Introduction and overview. PMID- 10823370 TI - A practical guide to the analysis of sirolimus using high-performance liquid chromatography with ultraviolet detection. AB - BACKGROUND: Sirolimus, a new immunosuppressive agent, has recently been approved in the United States for use in combination with cyclosporine and corticosteroids in renal allograft transplantation. Therapeutic drug monitoring (TDM) of sirolimus is advocated by the drug's manufacturer in certain patient populations. Given the known pharmacokinetic interaction of sirolimus with cyclosporine and the requirement for patient compliance, physicians may wish to monitor steady state trough levels of this agent. Several types of analytical methods have been investigated for use in TDM of sirolimus: immunoassay, high-performance liquid chromatography with ultraviolet detection (HPLC-UV), and HPLC with mass spectrometric detection (HPLC-MS or HPLC/MS/MS). OBJECTIVE: This review identifies analytical parameters that are critical to clinical TDM of sirolimus when HPLC-UV is used. METHODS: Extraction of sirolimus from whole blood was performed using either liquid-liquid or solid phase techniques, whereas liquid chromatography was performed on reverse-phase analytical columns. The drug was detected at its UV extinction maximum. Calculated analytical parameters were evaluated according to current industry standards. RESULTS: HPLC-UV methods for the quantification of sirolimus meet or exceed industry standards of performance for clinical TDM. Comparison of the sirolimus levels in clinical samples analyzed by both HPLC-UV and HPLC/MS/MS indicated that the methods provided similar results. CONCLUSION: HPLC-UV methods, although they use a less-sophisticated mode of detection than HPLC-MS or HPLC/MS/MS methods, provide an alternative means of performing TDM without sacrificing the analytical performance required for clinical monitoring of sirolimus. PMID- 10823371 TI - Therapeutic monitoring of sirolimus in human whole-blood samples by high performance liquid chromatography. AB - BACKGROUND: Sirolimus is a macrolide antibiotic isolated from Streptomyces hygroscopicus that has demonstrated immunosuppressive activity. Human and animal studies have shown a good correlation of trough sirolimus concentrations with immunosuppressive efficacy. OBJECTIVE: This report describes a reverse-phase high performance liquid chromatography (HPLC) method used for therapeutic drug monitoring of sirolimus. METHODS: A reverse-phase C18 column method was developed using an automated HPLC system and ultraviolet (UV) detection. Whole-blood samples collected in ethylenediamine-tetraacetic acid (EDTA) are first hemolyzed, and an internal standard (desmethoxysirolimus) is added to 1.0 mL of sample, which is then extracted with 1-chlorobutane and, after the organic layer is removed, evaporated to dryness. The residue is reconstituted in a 70% methanol/water mixture. Reconstituted extracts are analyzed by HPLC at a column temperature of 60 degrees C and a flow rate of 1.0 mL/min. Typically, chromatography requires 35 minutes between each sample injection. The UV detector is set at 278 nm with a response sensitivity of 0.010 AUFS (absorbance units full scale). Standards and controls prepared in hemolyzed EDTA-anticoagulated whole blood are extracted and run in parallel. Identification of peaks of interest is by retention time; quantification of sirolimus in controls and clinical samples uses a peak-height ratio (sirolimus/internal standard). RESULTS: The assay's precision (coefficients of variation, 5.7%-14.4%) and sensitivity (2.5 ng/mL) were found to be appropriate for therapeutic monitoring purposes. Analytical recovery of 88.0% to 106.3% was observed throughout the assay's linear range (2.5 150.0 ng/mL). Stability studies at 20 degrees C to 25 degrees C and 2 degrees C to 8 degrees C showed an estimated recovery of sirolimus ranging from 85% to 110% of target concentrations (10-90 ng/mL). In a study comparing the results of 194 samples from kidney transplant recipients assayed by the HPLC-UV assay and by a microparticle enzyme immunoassay, the HPLC-UV method provided approximately 10% lower values. CONCLUSION: The HPLC-UV assay is analytically capable of providing useful data for the clinical assessment of patients receiving sirolimus. PMID- 10823372 TI - Measurement of sirolimus in whole blood using high-performance liquid chromatography with ultraviolet detection. AB - BACKGROUND: Sirolimus, a potent immunosuppressive drug, exhibits intrapatient and interpatient variability of absorption and metabolism. Thus, therapeutic drug monitoring is important. OBJECTIVE: This paper describes a reverse-phase high performance liquid chromatography (HPLC) method, using ultraviolet (UV) absorption for detection, for measuring sirolimus levels in human whole-blood samples. METHODS: The stability of sirolimus in whole blood was assessed under conditions likely to be encountered during transport of study samples to a central laboratory. The performance of the HPLC-UV assay in measuring sirolimus was compared with that of 3 established, validated HPLC assays with tandem mass spectrometric (MS/MS) detection. Results of the HPLC-UV assay also were compared with results produced by a prototype microparticle enzyme immunoassay (MEIA). RESULTS: Inaccuracy for 3 in-house control samples was < or =4%, whereas within assay repeatability (coefficient of variation [CV]) was < or =5% and between assay reproducibility was < or =6.6%. Mean recovery of sirolimus from blood was 81.5%+/-4.3%. The lower limit of quantification was set at 6.5 ng/mL, and the repeatability CV at this concentration was 4.2% (n = 6). Sirolimus-containing whole-blood samples were stable for 3 freeze/thaw cycles when stored at -20 degrees C and for > or =2 days when stored at ambient temperature. The sample extract was shown to be stable for up to 54 hours at ambient temperature (approximately 22 degrees C) after extraction. Results of the HPLC-UV assay were consistent with those of the HPLC/ MS/MS assays but lower than those produced by MEIA. CONCLUSION: This HPLC-UV method is considered suitable for therapeutic drug monitoring of sirolimus. PMID- 10823373 TI - An immunoassay for the measurement of sirolimus. AB - OBJECTIVE: This study assessed the performance characteristics of a new microparticle enzyme immunoassay (MEIA) for the determination of sirolimus in whole blood. BACKGROUND: In clinical investigatory studies, dose adjustments of the immunosuppressive drug sirolimus have been carried out using either high performance liquid chromatography (HPLC) or, more recently, this investigational immunoassay kit based on the MEIA technique. METHODS: Calibration was made over the linear range 0 to 30 ng/mL. Inaccuracy and imprecision were assessed by means of 3 control samples supplied with the kit (5, 11, and 22 ng/mL) and dilution of an above-quantitation-limit sample (154 ng/mL). Specificity was determined by the addition of 2 sirolimus metabolites to sirolimus-free human whole blood or to I of the control samples supplied with the kit. In addition, whole-blood samples from patients receiving either cyclosporine or tacrolimus (N = 24) were analyzed for sirolimus. A comparison of the MEIA and a validated HPLC/MS/MS assay analyzed both pooled samples from patients receiving sirolimus and spiked samples (sirolimus 2-60 ng/mL). In a more extensive comparison of patient samples measured by the MEIA assay, a validated HPLC assay with UV detection (HPLC-UV) was used (HPLC-UV sirolimus 7-64 ng/mL). RESULTS: Inaccuracy (between-run) was < or =16.2% at all 4 concentrations (N = 5). Within-assay imprecision (repeatability) was <6% (N = 5), and between-assay imprecision (reproducibility) for the same samples was < 11% (N = 5). Recovery, assessed by means of 3 in-house control samples prepared in both fresh and previously frozen sirolimus-free human whole blood, ranged from 93.9% to 109.5%. The limit of detection, determined by dilution of the lowest nonzero calibrator (3 ng/mL), was set at 1 ng/mL, at which repeatability was 20.5% (N = 5). Five ng/mL of hydroxysirolimus cross-reacted with the assay by a mean of between 44% and 50% (N = 4); 5 ng/mL of 41-O demethylsirolimus cross-reacted with the assay by a mean of between 86% and 127% (N = 4). Assay specificity was further challenged by ethylenediamine-tetraacetic acid (EDTA)-whole-blood samples from transplant patients not receiving sirolimus. These samples had tacrolimus and cyclosporine concentrations of 7.8 to 15.9 ng/mL and 38 to 485 microg/L, respectively. The median result was 0 ng/mL (third quartile, 0.7 ng/mL; maximum, 1.4 ng/mL); no value was above the lowest nonzero calibrator. The results of the comparison between the MEIA and the HPLC/MS/MS assay showed mean positive biases of 21% and 8% for the MEIA in measuring sirolimus in pooled patient samples and spiked samples, respectively. The results of the comparison of the MEIA and HPLC-UV median sirolimus concentrations were 18.2 and 20.1. Whole-blood samples anticoagulated with EDTA and containing sirolimus were stable for analysis by MEIA for 3 freeze-thaw cycles when stored at -20 degrees C and for 10 days when stored at 4 degrees C or at ambient temperature. A decline in sirolimus concentration occurred when samples were stored at 37 degrees C. CONCLUSION: The MEIA showed suitable precision across a clinically relevant concentration range. In terms of patient management, the practical significance of cross-reactivity with sirolimus metabolites remains to be assessed. PMID- 10823374 TI - An immunophilin-binding assay for sirolimus. AB - OBJECTIVE: This review examines the performance of 4 assays for sirolimus in terms of their ability to meet 6 guidelines determined by a panel of experts. BACKGROUND: Four methods have been described to date for the analysis of sirolimus concentrations in whole blood: high-performance liquid chromatography mass spectrometry (HPLC-MS); microparticle enzyme immunoassay (MEIA); p70 S6 kinase inhibition; and an immunophilin-binding assay (IBA). METHODS: A MEDLINE search of the literature was performed to identify relevant studies. RESULTS: The HPLC methods suffer from precision problems because of the substantial specimen preparation required, and HPLC-MS methods are not practical for clinical use. Initial studies of the MEIA have found overestimation of sirolimus concentrations that may be caused by antibody cross-reactivity with sirolimus metabolites. Monitoring of sirolimus effects by p70 S6 kinase inhibition is as yet possible only theoretically, and the assay itself is not yet optimal. With the IBA, use of a T-cell protein that binds to sirolimus and that may be the intracellular target of the drug as the assay binding protein allows the assay to measure sirolimus selectively, even in the presence of structurally similar metabolites. CONCLUSION: More than 200 clinical samples have been analyzed by the IBA, and correlation with HPLC values has been good, with a regression line slope near 1.0. In addition, the assay is easier to perform and more precise than HPLC, and has the potential to be automated. Thus, the IBA appears to have certain clear advantages over the other assays. PMID- 10823375 TI - The quantification of sirolimus by high-performance liquid chromatography-tandem mass spectrometry and microparticle enzyme immunoassay in renal transplant recipients. AB - BACKGROUND: Sirolimus, an immunosuppressive agent, is undergoing clinical trials in the prophylaxis of organ rejection. OBJECTIVES: The aim of this study was to compare the performance of the semi-automated prototype (mode IA) microparticle enzyme immunoassay (MEIA) against a validated high-performance liquid chromatography-mass spectrometry (HPLC-MS) method for measuring sirolimus concentrations. A secondary objective was to identify potential factors that may influence sirolimus measurement. METHODS: The comparison was based on predose samples (n = 841) from 74 renal transplant patients receiving sirolimus therapy. Samples were collected up to 12 months after transplantation. RESULTS: The mean (+/- SD) overestimation by MEIA was 42.5%+/-16.9%. Several variables were investigated to determine potential contributors to the observed overestimation. Stratification of the data based on the mean sirolimus concentrations determined by both assays yielded no statistically significant differences in bias between concentration subgroups within the clinically relevant range. Multiple linear regression analysis identified HPLC-MS sirolimus concentration (P = 0.03), hemoglobin concentration (P < 0.001), and time after transplantation (P < 0.001) as significant variables in the prediction of overestimation by MEIA. Analysis of the effect of time after transplantation on overestimation yielded a statistically significant difference up to 6 months after transplantation (35.6% to 46.4%) compared with 9 (23.9%) and 12 months (24.4%). A relationship between hemoglobin concentration and time after transplantation may explain the reduction in bias observed after 6 months. CONCLUSION: The MEIA overestimates sirolimus concentrations in renal transplant patients compared with HPLC-MS. The clinical importance of this observed overestimation requires further investigation. PMID- 10823376 TI - Requirements for therapeutic drug monitoring of sirolimus, an immunosuppressive agent used in renal transplantation. AB - BACKGROUND: On September 15, 1999, sirolimus received approval from the US Food and Drug Administration (FDA) for marketing as an immunosuppressive agent. As with any chronically administered medication, the question arises whether therapeutic drug monitoring (TDM) is required for optimal therapy. In the case of sirolimus, there are data to suggest that TDM may be beneficial in some patients. OBJECTIVE: To assess the need for monitoring sirolimus concentrations, this paper reviews the following factors influencing the usefulness of TDM: wide pharmacokinetic variability; toxicity; suspected noncompliance; suspected drug interactions; and specific demographic characteristics. Data supporting the correlation between sirolimus concentration and immunosuppressive efficacy are also discussed. RESULTS: The available literature on sirolimus suggests that TDM may be required in some cases. Studies have shown that there is wide interindividual variability in the pharmacokinetic behavior of drugs in transplant patients; that there is a relationship between blood concentrations of sirolimus and adverse events; and that coadministration of cyclosporine alters the pharmacokinetics of sirolimus. Additionally, the correlation between sirolimus concentration and immunosuppressive efficacy in phase III trials suggests a benefit in transplant patients when sirolimus concentrations reach appropriate levels. Finally, noncompliance is a common occurrence in the transplant population, and monitoring is often necessary in suspected noncompliers. CONCLUSION: Although additional clinical studies are needed, it appears that TDM is an important aspect of treatment with sirolimus. PMID- 10823377 TI - Toxicity and efficacy of sirolimus: relationship to whole-blood concentrations. AB - BACKGROUND: Sirolimus is a novel macrolide immunosuppressive drug with a mechanism of action distinct from that of both cyclosporine and tacrolimus. Recent clinical studies have demonstrated a decrease in acute rejection episodes in renal transplant patients receiving sirolimus compared with controls. The major toxicities associated with sirolimus treatment are thrombocytopenia and hyperlipidemia. In addition, concern has been raised by the higher serum creatinine levels noted in patients receiving sirolimus and cyclosporine compared with controls receiving cyclosporine and azathioprine. OBJECTIVE: The objective of the present review is to summarize the efficacy and toxicity data for sirolimus. Special consideration is given to evidence that links these effects to dose or whole-blood concentrations of sirolimus. RESULTS: The literature indicates that trough concentrations of sirolimus >15 ng/mL appear to be associated with a greater risk of both thrombocytopenia and hyperlipidemia, whereas trough sirolimus concentrations <6 ng/mL have been associated with an increased incidence of acute rejection. CONCLUSION: The evidence to date supports target trough sirolimus concentrations of 6 to 15 ng/mL in most patients. In higher-risk groups and patients receiving cyclosporine-sparing regimens, higher concentrations may be necessary to achieve similar efficacy. PMID- 10823378 TI - Clinical pharmacokinetics and therapeutic drug monitoring of sirolimus. AB - BACKGROUND: Sirolimus is a novel macrocyclic antibiotic that has an immunosuppressive mechanism of action distinct from that of cyclosporine and tacrolimus. OBJECTIVE: The objective of this report is to provide an overview of the clinical development of sirolimus with emphasis on the mechanism of immunosuppressive activity, prevention of acute renal allograft rejection, clinical pharmacokinetics, concentration-effect relationships, and therapeutic drug monitoring (TDM). RESULTS: Pharmacokinetic studies in adult renal transplant patients have shown that sirolimus may be characterized as a drug with rapid absorption (t(max) = 1 to 2 hours), low systemic availability (F = 14%), linear dose proportionality (2 to 24 mg), extensive partitioning into formed blood elements (B/P = 36), large apparent volume of distribution (1.7 L/kg), prolonged terminal half-life (62 hours), and large intersubject (CV = 52%) and intrasubject (CV = 26%) variability in oral-dose clearance. Results from phase 111 pivotal trials showed that sirolimus (2 or 5 mg/d) reduced acute renal graft rejection (generally, P < 0.01) without TDM. Although TDM may not be required for a regimen consisting of full-dose cyclosporine and corticosteroids with sirolimus 2 mg/d (4 hours after cyclosporine), it may be warranted in patients (1) with hepatic impairment, (2) who are young children, (3) who are receiving concurrent doses of strong CYP3A/p-glycoprotein inhibitors or inducers, (4) in whom cyclosporine dosing is markedly reduced or discontinued, and (5) who are at a high risk for rejection. A whole-blood sirolimus therapeutic window of 5 to 15 ng/mL (measured by microparticle enzyme immunoassay) is recommended for patients at standard risk of rejection. The large intrapatient variability observed in trough sirolimus concentrations indicates that dose adjustments should be optimally based on more than a single trough sample. Because of the time required to reach steady state, sirolimus dose adjustments would optimally be based on trough levels obtained >5 to 7 days after a dose change. CONCLUSIONS: The effective use of sirolimus in an immunosuppressive regimen for the prevention of acute renal allograft rejection requires an understanding of the drug's clinical pharmacokinetics, concentration/adverse-effect relationship, concentration-efficacy relationship, and TDM. PMID- 10823379 TI - Proficiency-testing issues relating to sirolimus. AB - BACKGROUND: A need exists to document laboratory proficiency to (1) compare results produced by different analytical techniques and (2) ensure consistency of results from multiple testing sites. OBJECTIVES: To enable concentration controlled studies of sirolimus to proceed with confidence, proficiency-testing schemes were put in place at laboratories selected to act as reference laboratories. The feasibility of establishing an ongoing proficiency-testing scheme was addressed with respect to sample stability. The scheme was then used to test proficiency for the measurement of sirolimus in 3 blinded samples each month. METHODS: The method chosen for measurement of sirolimus was a prototype microparticle enzyme immunoassay. Initially, 15 laboratories were asked to analyze a series of 85 blinded samples that tested their inaccuracy, repeatability, and reproducibility for the measurement and their ability to dilute over-range samples competently. Both blood samples spiked with sirolimus and pooled blood samples from patients receiving the drug were circulated to a maximum of 50 laboratories. RESULTS: Overall, both inaccuracy and imprecision were acceptable by predefined criteria. Inaccuracy for the immunoassay (percentage difference of the measured value against the nominal value) averaged 5% (95% CI, -9% to -1%). The mean percentage difference between the immunoassay and a high-performance liquid chromatographic assay with mass-spectrometric detection for the measurement of sirolimus in pooled samples (n = 5) from patients receiving the drug was 29% (95% CI, 24% to 34%). CONCLUSION: The techniques documented here as part of the International Sirolimus Proficiency Testing Scheme could be applied to other clinical studies requiring protocol driven dosing adjustments based on sirolimus measurements, irrespective of analytical technique used. PMID- 10823380 TI - Assessing data elements in a severity scoring system. PMID- 10823381 TI - It is not nice to fool Mother Nature. PMID- 10823382 TI - Immunomodulation by anaesthetic, sedative and analgesic agents: does it matter? PMID- 10823383 TI - Using and understanding sedation scoring systems: a systematic review. AB - OBJECTIVE: To systematically review instruments for measuring the level and effectiveness of sedation in adult and pediatric ICU patients. STUDY IDENTIFICATION: We searched MEDLINE, EMBASE, the Cochrane Library and reference lists of the relevant articles. We selected studies if the sedation instrument reported items related to consciousness and one or more additional items related to the effectiveness or side effects of sedation. DATA ABSTRACTION: We extracted data on the description of the instrument and on their measurement properties (internal consistency, reliability, validity and responsiveness). RESULTS: We identified 25 studies describing relevant sedation instruments. In addition to the level of consciousness, agitation and synchrony with the ventilator were the most frequently assessed aspects of sedation. Among the 25 instruments, one developed in pediatric ICU patients (the Comfort Scale), and 3 developed in adult ICU patients (the Ramsay scale, the Sedation-Agitation-Scale and the Motor Activity Assessment Scale), were tested for both reliability and validity. None of these instruments were tested for their ability to detect change in sedation status over time (responsiveness). CONCLUSION: Many instruments have been used to measure sedation effectiveness in ICU patients. However, few of them exhibit satisfactory clinimetric properties. To help clinicians assess sedation at the bedside, to aid readers critically appraise the growing number of sedation studies in the ICU literature, and to inform the design of future investigations, additional information about the measurement properties of sedation effectiveness instruments is needed. PMID- 10823384 TI - Interobserver variability of the measurement of the mortality probability models (MPM II) in the assessment of severity of illness. AB - OBJECTIVE: To assess the interobserver variability of the measurement of the MPM II system. DESIGN: Random sample of an inception cohort. SETTING: Fifteen adult medical and surgical intensive care units (ICUs) in Spain. PATIENTS: A 5% random sample (n = 119) of 2332 patients consecutively admitted in the ICU, aged 18 years or older. INTERVENTIONS: Prospective data collection during the stay of the patient in the ICU. Repeated data collection, after the patient's hospital discharge. MEASUREMENTS AND MAIN RESULTS: Demographic information, length-of-stay and vital status at both ICU and hospital discharge, as well as all variables necessary for computing the MPM II system were measured. Interobserver variability for categorical variables was measured computing the kappa index. For interval variables the interobserver variability was assessed by the intraclass correlation coefficient, the paired t-test, and linear regression. In the MPM II0 index, the variables coma, heart rate, systolic blood pressure, chronic renal insufficiency, metastatic neoplasm, acute renal failure, cardiac dysrhythmia and cardiopulmonary resuscitation, showed moderate or low levels of agreement. In the MPM II24 index, the variables coma or deep stupor, PaO2 and prothrombin time showed low or moderate interobserver agreement. Agreement for the MPM II probability of death was high, although the external observer obtained a lower estimation than the hospital observer. CONCLUSIONS: The MPM II severity system showed a high level of stability when used in groups of patients. Nevertheless, the observed variability in some variables means that it should be used with caution for individual patients. PMID- 10823385 TI - Angiotensin II formation and endothelin clearance in ARDS patients in supine and prone positions. AB - OBJECTIVE: In patients with acute respiratory distress syndrome (ARDS), the prone position may enhance oxygenation by changing ventilation/perfusion ratio. In this study, we investigated whether the prone position affects the net balance between pulmonary endothelin (ET-1) and angiotensin II (Ang II) production and clearance, two metabolic functions of lung endothelial cells. SETTING: Anaesthesiological intensive care unit of a university hospital. PATIENTS: Ten ARDS patients (Murray score > 2.5) were studied in both the supine position (SP) and the prone position (PP). MEASUREMENTS AND DESIGN: Blood samples were taken simultaneously from the patient in SP for assessment of mixed venous and arterial ET-1 and Ang II concentrations, and plasma renin concentration (PRC). This was repeated after 60 min in SP, immediately after turning the patient into PP, and 60 min thereafter. Net arterial/mixed venous ET-1 clearances and net Ang II formations were calculated. RESULTS: arterial oxygen tension increased from SP to PP by an average of 60 mmHg, about 20%. Arterial ET-1 concentrations of ARDS patients were 1.57 +/- 1.1 pg/ml (mean +/- SD) and within the range of healthy persons. Net ET 1 clearances were negative in SP, indicating pulmonary release of ET-1, and did not change in PP. Arterial Ang II concentrations (73 +/- 56 pg/ml) as well as PRC (126 +/- 85 pg/ml) were markedly elevated. Net transpulmonary Ang II formation did not change. CONCLUSION: Acute changes of oxygenation in ARDS patients by positioning do not induce any short-term effects on pulmonary ET-1 net clearance or Ang II net formation. PMID- 10823386 TI - Could anti-inflammatory actions of catecholamines explain the possible beneficial effects of supranormal oxygen delivery in critically ill surgical patients? AB - OBJECTIVE: To evaluate the literature regarding antiinflammatory actions of cytokines, evaluate randomized controlled trials (RCTs) of supranormal oxygen delivery, and suggest alternative mechanism(s) for possible beneficial effects of supranormal oxygen delivery in critically ill surgical patients. DESIGN: Literature review using Medline and review of selected illustrative studies. MAIN RESULTS: Catecholamines (epinephrine, norepinephrine, isoproterenol, and dopamine) in general inhibit tumor necrosis factor-alpha (TNF) production and may enhance interleukin-6 (IL-6) and IL-10 production. Phosphodiesterase inhibitors also inhibit TNF and may enhance IL-10. All studies used models (cell, animal, or humans infused with endotoxin) of sepsis. RCTs of supranormal oxygen delivery show decreased mortality in high-risk surgical patients; however, prevention or reversal of tissue hypoxia may not be the mechanism of benefit. Antiinflammatory effects of catecholamines are a potential and, to date, unexplored mechanism of the benefit of supranormal oxygen delivery in critically ill surgical patients. CONCLUSIONS: Catecholamines may modulate cytokine response beneficially and could be a mechanism of decreased morbidity and mortality of supranormal oxygen delivery in high-risk surgical patients. PMID- 10823387 TI - Circulating endothelin-1 and tumor necrosis factor-alpha: early predictors of mortality in patients with septic shock. AB - OBJECTIVES: To determine the predictive value of early determination of tumor necrosis factor (TNF)-alpha, TNF-alpha 1 and 2 soluble receptors (sTNFR1 and sTNFR2) and endothelin-1 (ET-1) for mortality in patients with septic shock. DESIGN: Prospective study. SETTING: Intensive care unit of a university hospital. PATIENTS: Twenty-one patients with septic shock. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: Patients with septic shock had a pulmonary artery catheter inserted and blood samples drawn at time zero, 6, 12 and 24 h, simultaneously with hemodynamic assessments. Plasma levels of all markers were measured by ELISA. All patients were followed up to hospital discharge or death. Age and APACHE II scores were significantly higher in nonsurvivors (n = 11) than in survivors (n = 10). Hemodynamic assessments did not aid in the discrimination between the two groups of patients (P > 0.05). Levels of TNF-alpha were higher in nonsurvivors than in survivors at all time-points. sTNFR1 and sTNFR2 were also significantly elevated in nonsurvivors, but not in all measurements. Endothelin 1, however, was significantly higher in nonsurvivors than in survivors only at 6 h (P = 0.02). When both TNF-alpha and ET-1 were increased at early time-points, the best predictive values for mortality were obtained [positive and negative predictive values of 72 and 100% at 6 h, odds ratio 3.0, 95% CI (1.2-7.6)]. CONCLUSIONS: Increased levels of TNF-alpha were consistently higher at all time points in nonsurvivors with septic shock. ET-1 levels, however, appeared also to be an early and sensitive predictor of mortality. Very early determination of TNF alpha and ET-1 in septic shock may help to identify patients at higher risk for adverse outcome. PMID- 10823388 TI - Early hyperlactataemia in critically ill children. AB - OBJECTIVE: To examine the relationships between early hyperlactataemia, acidosis, organ failure, and mortality in children admitted to intensive care. DESIGN: Prospective observational study. Children with lactate levels > 2 mmol/l were eligible for enrolment. Post-operative patients and those with inherited metabolic disease were excluded. Seven hundred and five children admitted to intensive care were screened, and 50 children with hyperlactataemia (incidence 7%), aged 20.3 months (0.1-191) were enrolled and followed up. The Paediatric Risk of Mortality (PRISM) score, Multiorgan System Failure (MOSF) score, length of ICU stay, and outcome were recorded. Data were collected for lactate (mmol/l), pH, and base excess (BE) until 24 h after admission. Data are reported as median (range) and were analysed by the Mann-Whitney, Fisher's Exact, and Kruskal-Wallis tests, and chisquared test for trend. RESULTS: Overall mortality in the screening group was 70/705 (10%). In the study group (n = 50) median PRISM score was 19 (4 49), median MOSF score 2 (1-4), and observed mortality 32/50 (64%). Median duration of ICU stay was 6 days (2-32) in survivors, and median time until death 3 days (0-13) in nonsurvivors. Eleven nonsurvivors (34%) died within 24 h. In the screening group, hyperlactataemia on admission identified mortality with likelihood ratio = 15. In the study group, neither the admission lactate (3.8 vs 4.6 mmol/l, P = 0.27), pH (7.32 vs 7.30, P = 0.6), nor BE (-7.5 vs -8, P = 0.45) differed significantly between survivors and nonsurvivors. Neither the admission nor peak lactate increased with increasing MOSF score (P = 0.5 and 0.54). The median peak lactate level was 5 mmol/l (2-9.3) in survivors compared to 6.8 mmol/l (2.3-22) in nonsurvivors (P = 0.02), and the cumulative average lactate level was 2.4 mmol/l (1-4.9) in survivors, compared to 4.5 mmol/l (1.6-21) in nonsurvivors (P = 0.0003). Persistent hyperlactataemia 24 h after admission identified mortality with likelihood ratio = 7. CONCLUSION: Hyperlactataemia on admission to intensive care is associated with a high mortality in children. Nonsurvivors within this group may be distinguished by the peak lactate level, or by persistent hyperlactataemia after 24 h of treatment. PMID- 10823389 TI - Oral health status of children in a Paediatric Intensive Care Unit. AB - OBJECTIVE: The aim of the present study was to examine the dental status of critically ill children in a Paediatric Intensive Care Unit (PICU) and determine the efficacy of the mouth care received. DESIGN: Prospective nonrandomized study. SETTING: The study was carried out in the PICU at Great Ormond Street Hospital for Children. PATIENTS AND PARTICIPANTS: All children admitted to the PICU during the period of the study were eligible for inclusion. A total of 54 children completed the study. MEASUREMENTS AND RESULTS: Children were examined for dental caries, plaque accumulation, gingival inflammation and gingival bleeding on admission to the PICU. The examination was repeated on discharge from the Unit. The levels of dental caries found on examination were compared with results from the most recent national surveys carried out in the United Kingdom, and analysed using the one sample t-test. The levels of plaque accumulation, gingival inflammation and gingival bleeding on admission were compared to those on discharge and analysed using the paired sample t-test. Results revealed a very highly significant increase in plaque accumulation (p = 0.001), and a highly significant increase in gingival inflammation (p = 0.006) between admission to the PICU and discharge. CONCLUSIONS: Results indicate that the present mouth care regimen is not effective in preventing the build up of plaque or maintaining gingival health. These children may therefore be at unnecessary risk from local or systemic spread of oral microorganisms. PMID- 10823390 TI - The CRIB (Clinical Risk Index for Babies) score and neurodevelopmental impairment at one year corrected age in very low birth weight infants. AB - OBJECTIVE: To assess the ability of the Clinical Risk Index for Babies (CRIB) to predict long-term neurodevelopmental impairment in very low birth weight (VLBW) infants. DESIGN: Single-center cohort study. SETTING: Tertiary neonatal care hospital and follow-up clinic. PATIENTS: Four hundred fifty-five VLBW infants consecutively admitted from 1992 to 1997 inclusive. MEASUREMENTS AND RESULTS: Calculations of CRIB scores from birth weight, gestational age, the presence of congenital malformations, worst base excess, maximum and minimum appropriate fraction of inspired oxygen (FIO2) during the first 12 h of life was possible in 430 infants. Three hundred eighty-six infants survived until discharge (89%) and 352 (91%) were examined at 1 year corrected age using the Griffiths scales of mental development. Major neurodevelopmental impairment (general quotient < 2 standard deviations below average) was observed in 76 infants (22%). CRIB scores and the individual CRIB components differed significantly between infants with and those without neurodevelopmental impairment. By logistic regression analysis, CRIB scores and minimum FIO2 were independent predictors of death, while CRIB and maximum FIO2 were independently associated with neurodevelopmental impairment. For combined poor outcome (death or impairment), CRIB, minimum and maximum FIO2 were independent predictors. In predicting major neurodevelopmental impairment, the area under the receiver operating characteristic curve for CRIB (0.703 +/- 0.035) did not differ significantly from that of birth weight (0.697 +/- 0.035) or any other CRIB component. CONCLUSION: While high CRIB scores are associated with major neurodevelopmental impairment, the CRIB score is of limited value for stratification in randomized trials or for adjustments in comparing performance between hospitals with neurodevelopmental impairment as the main outcome measure. PMID- 10823391 TI - Recovery from withdrawal of inhaled nitric oxide and kinetics of nitric oxide induced inhibition of nitric oxide synthase activity in vitro. AB - OBJECTIVE: To examine the kinetics of successful nitric oxide (NO) withdrawal in vivo and in vitro. DESIGN AND SETTING: Prospective study in a university pediatric intensive care ward and research laboratory. PATIENTS AND MATERIALS: Nineteen patients with acute respiratory distress syndrome (ARDS) or persistent pulmonary hypertension of the newborn (PPHN). Primary porcine pulmonary artery cells in vitro. INTERVENTIONS: NO inhalation and withdrawal in patients; exposure to NO donor sodium nitroprusside (SNP) and gaseous NO in vitro. MEASUREMENTS AND RESULTS: In patients: a slight, but significant, increase of oxygenation index (OI) from 4.57 +/- 0.24 cmH2O/torr (mean +/- SEM) to 4.90 +/- 0.26 cmH2O/torr after withdrawal of NO (p < 0.001). Recovery of OI (4.43 +/- 0.23 cmH2O/torr) 30 min after weaning, a significant drop after 4 h (3.72 +/- 0.17 cmH2O/ torr;p < 0.001), values restored after 12 h. In vitro: NO synthase (NOS) activity was significantly lower in SNP-incubated cells (20.0 +/- 4.0 microM/min) than in control cells (37.6 +/- 7.0 microM/ min; p < 0.05). Thirty minutes after SNP withdrawal there was NOS activity of 35.8 +/- 10.0 microM/min with a significant increase by 4 h (p < 0.05). No alteration of endothelial NOS (ENOS) mRNA expression by NO (Northern Blot). CONCLUSION: In patients there is a slight, but significant, reversible increase of OI after successful weaning from NO. In vitro, NO leads to a reversible decrease of ENOS activity on a post mRNA level, resembling clinical observations. PMID- 10823392 TI - Effects of NO inhalation on pulmonary leukocyte sequestration and blood volume in porcine endotoxaemia. AB - OBJECTIVE: Sequestration and migration of activated neutrophils plays a major role in the pulmonary injury typical of septic shock and the adult respiratory distress syndrome. Inhaled NO may counteract alveolar-capillary damage attributed to activated neutrophils. The present study describes a method to directly demonstrate the effects of NO inhalation on endotoxin-induced sequestration of 99mTc-labelled leukocytes [As(t)] in the lungs of pigs. DESIGN: Prospective controlled study. SETTING: Laboratory for experimental surgery at a university medical centre. SUBJECTS: Anaesthetised and ventilated pigs. INTERVENTIONS: To induce inflammatory shock 26 animals received a continuous endotoxin infusion. Thirteen animals inhaled NO from the start of the experiments, while 13 served as controls. In 13 animals from both groups, leukocytes were labelled in vitro and reinjected, while in the 13 others erythrocytes were labelled in vivo to provide corrections for changes in blood volume. MEASUREMENTS AND RESULTS: The pulmonary distribution of 99mTc-labelled leukocytes or erythrocytes was studied dynamically for 180 min. After correction for changes in pulmonary and heart blood volume (PBV, HBV), leukocyte sequestration curves were generated. Endotoxin induced pulmonary vasoconstriction, reduced PBV, impaired oxygenation, and caused a maximum increase in As(t) of 30% in the lungs. NO inhalation attenuated pulmonary vasoconstriction and the reduction in PBV. The maximum increase in As(t) was reduced to 15% of baseline. CONCLUSIONS: Inhaled NO exerts its main vascular effects in the pulmonary microvasculature, the primary site of physiological neutrophil margination and pathological adhesion of activated leukocytes. Early use of NO inhalation may offer protection against the development of more lasting pulmonary failure in septic shock by reducing leukocyte sequestration in the lungs. PMID- 10823393 TI - Red blood cell transfusion in the intensive care unit. PMID- 10823394 TI - Substance abuse and acute asthma. PMID- 10823395 TI - Mechanical circulatory support in pediatric patients. PMID- 10823396 TI - Acute postrenal failure and pseudomyocardial infarction after spontaneous bladder rupture. PMID- 10823397 TI - Hyperbaric oxygen delivery in treatment of a child with Klippel-Trenaunay syndrome complicated with a limb threatening Escherichia coli infection. PMID- 10823398 TI - Mechanical ventilation and intra-aortic counterpulsation in cardiogenic shock. PMID- 10823399 TI - A comparison of the abuse liability and dependence potential of nicotine patch, gum, spray and inhaler. AB - RATIONALE: Nicotine replacement therapy (NRT) in varying forms is becoming widely used. Clinicians, therapists and regulatory authorities are interested in the abuse liability and dependence potential of the different forms. OBJECTIVES: To compare the abuse liability and dependence potential of nicotine gum, transdermal patch, spray and inhaler. METHODS: 504 male and female smokers seeking help with stopping smoking were randomly allocated to the four products. Measures were taken at the designated quit date, then 1 week, 4 weeks, 12 weeks and 15 weeks later. Smokers were advised to use the product for up to 12 weeks. Those still using the product at the 12-week visit were advised to cease use by week 14. Measures included: pleasantness and satisfaction ratings at weeks 1 and 4 (used as a marker of abuse liability); ratings of feeling dependent on NRT at weeks 1, 4, 12 and 15 (used as a marker of subjective dependence); mood and physical symptoms ratings at weeks 12 and 15 (the change being used to assess physical dependence on NRT), continued usage of NRT at week 15 (used as an marker of behavioural dependence). RESULTS: Average ratings of pleasantness were low. The nicotine patch was rated as less unpleasant to use than all other products. There were no significant differences between the products in terms of satisfaction or subjective dependence except at week 15 when no patch users rated themselves as dependent. Continued use of NRT at week 15 was related to rate of delivery of nicotine from the products - 2% for patch, 7% for gum and inhaler, 10% for spray (P<0.05 for linear association). Among those PMID- 10823400 TI - A naturalistic investigation of the effects of day-long consumption of tea, coffee and water on alertness, sleep onset and sleep quality. AB - RATIONALE: The effects of caffeine, especially caffeinated coffee, on human performance have been extensively studied. However, few studies have been naturalistic representations of how tea/coffee is normally consumed in terms of dose and time of consumption. OBJECTIVES: This study investigated the effects of day-long consumption of tea, coffee and water on cognitive and psychomotor performance, and sleep quality at night. METHODS: Thirty healthy volunteers received equal volume drinks equivalent to either 1 or 2 cups of tea (containing 37.5 mg or 75 mg caffeine), or coffee (75 mg or 150 mg caffeine), or water, in a randomised five-way crossover design. Drinks were administered on four occasions during the day (0900, 1300, 1700 and 2300 hours). A psychometric battery consisting of critical flicker fusion (CFF), choice reaction time (CRT) and subjective sedation (LARS) tests, was administered pre-dose and at frequent time points post-dose. The Leeds Sleep Evaluation Questionnaire (LSEQ) was completed each morning and a wrist actigraph was worn for the duration of the study. RESULTS: Caffeinated beverages maintained CFF threshold over the whole day (P<0.05), independent of caffeine dose or beverage type. During the acute phase of beverage ingestion, caffeine significantly sustained performance compared to water after the first beverage for CFF and subjective sedation (P<0.05), and after the second beverage for the Recognition component of the CRT task (P<0.05). Additionally, there were significant differences between tea and coffee at 75 mg caffeine after the first drink. Compared to coffee, tea produced a significant increase in CFF threshold between 30 and 90 min post-consumption (P<0.01). However, following the second beverage caffeinated coffee at 75 mg significantly improved reaction time (P<0.05), compared to tea at the same dose, for the Recognition component of the CRT task. Caffeinated beverages had a dose dependent negative effect on sleep onset (P<0.001), sleep time (P<0.001) and sleep quality (P<0.001). CONCLUSIONS: These results indicate that ingestion of caffeinated beverages may maintain aspects of cognitive and psychomotor performance throughout the day and evening when caffeinated beverages are administered repeatedly. This study also demonstrates that day-long tea consumption produces similar alerting effects to coffee, despite lower caffeine levels, but is less likely to disrupt sleep. Other differences between tea and coffee were more subtle, and require further investigation. PMID- 10823401 TI - Regional heterogeneity for the intracranial self-administration of ethanol within the ventral tegmental area of female Wistar rats. AB - RATIONALE: Because current findings indicate that the selectively bred alcohol preferring P line of rats self-administers 50-200 mg% ethanol (EtOH) directly into the ventral tegmental area (VTA), whereas the alcohol-nonpreferring NP line does not, it is important to determine whether unselected, common stock rats would self-administer EtOH directly into the VTA. In addition, because neuroanatomical and self-administration studies indicate that the VTA may be functionally heterogeneous, the present study was designed to determine whether there were subregional differences within the VTA for the intracranial self administration (ICSA) of EtOH. OBJECTIVES: The objective of this study was to employ the ICSA technique to determine whether adult female Wistar rats would self-administer EtOH directly into the VTA, and whether regional heterogeneity existed for EtOH self-infusion within the VTA. METHODS: Following surgery to implant guide cannulae aimed at either the posterior or anterior VTA, subjects were placed in standard experimental chambers equipped with an 'active lever' [fixed ratio (FR)1 schedule of reinforcement], which caused the delivery of the infusate, and an 'inactive lever', which had no programmed consequence. Subjects were assigned to groups that self-administered either artificial cerebrospinal fluid (aCSF) throughout, or 100-400 mg% EtOH for the first four sessions (acquisition), aCSF in sessions 5 and 6 (extinction), and EtOH again during session 7 (reinstatement). RESULTS: During the four acquisition sessions, rats with posterior VTA placements readily self-administered 200 mg% and 250 mg% EtOH and discriminated between the active and inactive levers. These subjects also demonstrated extinction, when aCSF was substituted for EtOH, and reinstatement when EtOH was reintroduced. Rats with posterior VTA placements self-infused 300 mg% and 400 mg% EtOH, and demonstrated lever discrimination only during the initial acquisition sessions. In contrast, rats with anterior VTA placements did not self-administer EtOH. CONCLUSIONS: The findings suggest that EtOH is reinforcing within the posterior VTA of Wistar rats, and the VTA is a functionally heterogeneous structure with regard to EtOH reinforcement. PMID- 10823402 TI - Effects of the non-competitive NMDA-receptor antagonist memantine on morphine- and cocaine-induced potentiation of lateral hypothalamic brain stimulation reward. AB - RATIONALE: NMDA-receptor antagonists may be of potential therapeutic use in several states of disease. It has been reported that drugs like MK-801 can potentiate the rewarding effects of other drugs, which may complicate the therapeutic use of this class of drugs. However, since MK-801 appears to be an "atypical" drug in several respects, other NMDA-receptor antagonists may not share this effect of MK-801. OBJECTIVES: We tested the effects of memantine, a clinically used NMDA-receptor antagonist, in a paradigm that has previously shown the reward-potentiating effects of MK-801 to see if this drug would yield qualitatively comparable results. METHODS: The effects of memantine on morphine- and cocaine-induced potentiation of brain stimulation reward were examined, using the rate-free curve-shift paradigm. RESULTS: Low doses of morphine (2.5 mg/kg) and cocaine (5 mg/kg) produced moderate decreases in the reward threshold frequency reflected in moderate leftward shifts of the function relating response rate to stimulation frequency. These effects were not altered by co administration of an intermediate dose of memantine (5 mg/kg), but maximum response rate was significantly increased by these drug combinations. Higher doses of morphine (7.5 mg/kg) and cocaine (10 mg/kg) had stronger effects on the rate-frequency function and reward threshold. These effects were enhanced by co administration of a high dose of memantine (10 mg/kg), while the effects on maximum response rate were less pronounced. CONCLUSIONS: These results demonstrate that fairly high doses of memantine and morphine or cocaine have to be combined in order to observe an enhancement of the latter drugs' potentiation of brain stimulation reward. In this respect, memantine differs markedly from MK 801, another non-competitive NMDA-receptor antagonist which has been shown to interact with morphine and cocaine at very low doses. PMID- 10823403 TI - Sex differences in nicotine substitution to a pentylenetetrazol discriminative stimulus during ethanol withdrawal in rats. AB - RATIONALE: Nicotine and ethanol are frequently co-abused in men and women, but few studies compare common stimulus effects produced by these substances between males and females. OBJECTIVES: This study compared the anxiety-like behavior induced by nicotine prior to and during ethanol withdrawal in intact male, sham operated female, and ovariectomized (OVX) rats. METHODS: Using an animal model of anxiety, the pentylenetetrazol (PTZ) drug-discrimination assay, rats were trained to discriminate PTZ (16 mg/kg, i.p.) from saline and were subjected to the following tests: (1) PTZ-lever selection at 12 h after termination of ethanol diet (4.5% for 10 days); (2) dose-response tests for nicotine (0.08-1.3 mg/kg) prior to ethanol and 1.5, 6, and 7 days after ethanol withdrawal. RESULTS: (1) During acute ethanol withdrawal (12 h), more male rats (43.4%) responded on the PTZ lever than OVX (29%) or sham female (15.3%) rats. (2) For nicotine dose response tests, more male rats (70%) selected the PTZ lever than OVX (37.5%) or sham female (50%) rats prior to ethanol. At 1.5 days, nicotine fully generalized to the PTZ stimulus in male (100%) and OVX (90%), but only partially in sham female (50%) rats. At 6 days and 7 days after ethanol withdrawal, the PTZ-lever selection decreased, but more male rats (78%) tended to respond on a PTZ lever than OVX (63.6%) or sham female rats (62.5%). CONCLUSIONS: Acute nicotine produces anxiety-like behavior similar to that of PTZ in male and female rats, and this effect of nicotine is intensified during ethanol withdrawal in male and OVX rats, but not in sham female rats. PMID- 10823404 TI - Effects of zopiclone on the rest/activity rhythm after a westward flight across five time zones. AB - The effects of zopiclone on the rest/activity rhythm were investigated after a westward flight with a 5-h phase delay (Grenoble-Martinique). The rest/activity rhythm was recorded continuously with an actigraph monitor during a 6-day baseline episode in Grenoble and a 6-day sojourn in Martinique. Rectal temperature was recorded before the flight on day 2 and after the flight on days 2 and 5. Subjective jet-lag score was assessed before the flight on days 1 and 2 and in Martinique on days 1, 2, 5 and 6. Zopiclone (7.5 mg) or placebo were administered double blind 30 min prior to bedtime during the first 4 post-flight days (D1-D4). Under zopiclone treatment, sleep was less fragmented on the first post-flight night and the curtailment of sleep duration observed on D2 and D3 under placebo was prevented. During day-time, subjects tended to be more active than with placebo. By contrast, subjective jet-lag scores did not differ between both groups. Zopiclone seemed to accelerate the readjustment of the rest/activity rhythm and the normalisation of the phase relationship between sleep and the temperature rhythm. Zopiclone probably exerts its "resetting" effects on jet-lag desynchronosis by facilitating sleep induction rather than via a chronobiotic action. PMID- 10823405 TI - Prolactin response to nemonapride, a selective antagonist for D2 like dopamine receptors, in schizophrenic patients in relation to Taq1A polymorphism of DRD2 gene. AB - The dopamine D2 receptor (DRD2) gene has a Taq1A restriction fragment length polymorphism yielding two alleles, A1 and A2. It has been shown that the subjects with less frequent allele, the A1 allele, have lower density and diminished function of DRD2 in the striatum, compared to those with no A1 allele. In the present study, the relationship between this polymorphism and prolactin response to nemonapride, an antipsychotic drug with selective and potent DRD2 antagonistic property, was investigated in 25 Japanese schizophrenic inpatients (13 males, 12 females). The daily dose of nemonapride was fixed at 18 mg, and the duration of treatment was 3 weeks. Taq1A genotypes were determined by the polymerase chain reaction method. Plasma prolactin concentrations were measured by enzyme immunoassay. The subjects were divided into four subgroups by gender and Taq1A genotypes, i.e., six males and eight females with the A1 allele, seven males and four females with no A1 allele. The (delta)prolactin (change from the pretreatment concentration) at 1 week was significantly (P<0.05) higher in females with the A1 allele (78.0 +/- 47.1 ng/ml) than in males with the A1 allele (33.4 +/- 14.0 ng/ml) or with no A1 allele (29.5+/-24.8 ng/ml). In addition, (delta)prolactin at 3 weeks was significantly (P<0.05) higher in females with the A1 allele (98.1+/-67.9 ng/ml) than in females with no A1 allele (33.4+/-24.6 ng/ml), males with the A1 allele (29.1+/-17.3 ng/ml) or males with no A1 allele (28.6+/-22.0 ng/ml). The present study thus suggests that female patients with the A1 allele show a greater prolactin response to nemonapride, who may have a high risk for adverse effects associated with neuroleptic-induced hyperprolactinemia. PMID- 10823406 TI - d-Fenfluramine in panic disorder: a dual role for 5-hydroxytryptamine. AB - RATIONALE: 5-Hydroxytryptamine (5-HT) appears to modulate different forms of anxiety in different ways, but the importance of this in human anxiety disorders is unknown. OBJECTIVES: To investigate whether the 5-HT releasing agent d fenfluramine (dFEN) has different effects on resting and panic anxiety in panic disorder. METHODS: Thirteen drug-free patients with DSM-IIIR panic disorder were tested in a double-blind placebo-controlled crossover design. Carbon dioxide 7% (CO2) was given as a panic challenge 270 min after administration of dFEN or placebo. RESULTS: Compared to placebo, dFEN increased anxiety and arousal, maximal at 120 min, but tended to reduce CO2-induced anxiety and panic attacks with a significant reduction in Panic Visual Analogue Scale ratings (P=0.040). Anxiety following CO2, but not dFEN, administration resembled panic attacks (compared to Acute Panic Inventory symptom profile during patients' usual attacks). Patients with more severe disorders exhibited enhanced behavioural responses and blunted prolactin responses to dFEN. CONCLUSIONS: dFEN caused anxiety similar to generalised anxiety in panic disorder patients but reduced anxiety following 7% CO2 challenge, a laboratory analogue of naturally occurring panic attacks. These findings are consistent with a dual role for 5-HT in pathological anxiety. Patients with more severe symptoms differed in 5-HT function compared to more mildly affected individuals. PMID- 10823407 TI - The 5-HT(1A) receptor agonist 8-OH-DPAT reduces rats' accuracy of attentional performance and enhances impulsive responding in a five-choice serial reaction time task: role of presynaptic 5-HT(1A) receptors. AB - RATIONALE: Whilst several studies have investigated the role of serotonergic receptor subtypes in learning and memory, relatively few studies have examined their role in attentional processes. OBJECTIVE: The present study investigated the role of pre- and postsynaptic 5-HT1A receptors on rats' attentional performance in the five-choice serial reaction time task (5-CSRT). METHODS: Hungry rats were trained in the 5-CSRT task to detect brief (0.5 s) flashes of light presented randomly in one of five locations with a fixed intertrial interval of 5 s paced by the rat. We studied the effects of 8-OH-DPAT, a 5-HT1A receptor agonist, at various subcutaneous (SC) doses (10-100 microg/kg) on measures of rats' discriminative accuracy (the index of attentional functioning) and various behavioural indices of response control and motivation. Manipulations of basic task parameters, intracerebroventricular (ICV) injections of 5,7 dihydroxytryptamine (5,7-DHT) to deplete forebrain 5-HT and treatments with a selective 5-HT1A receptor antagonist WAY 100635 were made in order to determine the behavioural and neural specificity of the effects of 8-OH-DPAT. RESULTS: A dose of 100 microg/kg, but not lower doses, significantly reduced choice accuracy and increased errors of omission, latencies to respond correctly and to collect food reward and premature responses. All these effects were completely blocked by WAY 100635, injected SC 5 min before 8-OH-DPAT at doses from 10-100 microg/kg. WAY 100635 by itself had no effect in the task. Dimming the visual stimuli to one third of the usual brightness did not modify the effect of 8-OH-DPAT on choice accuracy. Prolonging the stimuli from 0.5 to 1.0 s reversed 8-OH-DPAT's effect on choice accuracy but did not modify the other effects on rats' performance. An ICV injection of 150 microg 5,7-DHT, which depleted forebrain serotonin by 90%, reversed 8-OH-DPAT's effect on choice accuracy but did not modify the effects on errors of omission and latency to make correct responses. Similar effects were found by infusing 1.0 microg/0.5 microl WAY 100635 in the dorsal raphe 5 min before 8-OH-DPAT. 8-OH-DPAT increased the latency to collect the reinforcement; this effect was attenuated by ICV 5,7-DHT and completely antagonized by WAY 100635 in the dorsal raphe. Rats treated with 5,7-DHT or 8-OH-DPAT showed more premature responses and these effects were markedly reduced by the combined treatment. CONCLUSIONS: The results suggest that stimulation of presynaptic 5 HT1A receptors is involved in the ability of 8-OH-DPAT to cause attentional dysfunction and enhance impulsivity while slowing of responding and increase in errors of omission mainly depend on stimulation of post-synaptic 5-HT1A receptors. PMID- 10823408 TI - Neuroactive steroids attenuate cocaine-induced sucrose intake in rats, but not cocaine-induced hyperactivity in mice. AB - RATIONALE: Neuroactive steroids, including the potent anticonvulsants ganaxolone (3alpha-hydroxy-3beta-methyl-5alpha-pregnan-20-one) and Co 2-1068 (3beta-(4acetyl phenyl)ethynyl-3alpha,21-dihydroxy-5beta+ ++-20-one-21-hemisuccinate), have recently been shown to protect against cocaine-induced seizures. OBJECTIVES: The purpose of the present experiments was to determine whether ganaxolone and Co 2 1068 attenuate acute behavioral effects of cocaine unrelated to seizures. METHODS: In the first experiment, the locomotor effects of Co 2-1068 (10-100 mg/ kg), pentobarbital (10-100 mg/kg) and haloperidol (0.03-0.3 mg/kg), alone or in combination with cocaine (5.6-30 mg/kg), were determined in mice. In the second experiment, the effects on sucrose intake of ganaxolone (4-16 mg/kg), Co 2-1068 (8-64 mg/kg), pentobarbital (4-32 mg/kg), and haloperidol (0.04-0.4 mg/kg), alone or in combination with cocaine (4-16 mg/kg), were determined in rats. RESULTS: Cocaine caused a dose-related increase in locomotor activity in mice, whereas Co 2-1068, pentobarbital and haloperidol caused dose-related decreases. The dopamine antagonist haloperidol, at a dose that had no effect on activity by itself, but not Co 2-1068 or pentobarbital, attenuated the cocaine-induced increase in locomotor activity. Cocaine, ganaxolone, Co 2-1068, and haloperidol produced dose related decreases in sucrose intake in rats; the effects of pentobarbital on sucrose intake were variable. As with locomotor effects, haloperidol attenuated the cocaine-induced decrease in sucrose intake. In addition, cocaine-induced decreases in sucrose intake were attenuated by ganaxolone and Co 2-1068. Pentobarbital had no statistically significant effect on the cocaine dose response function. CONCLUSIONS: These results suggest that the interaction of neuroactive steroids with cocaine extends to pharmacologic actions beyond anticonvulsant efficacy, but that the blockade of behavioral effects of cocaine by neuroactive steroids does not apply to all acute behaviors. PMID- 10823409 TI - Distress vocalizations in maternally separated mouse pups: modulation via 5 HT(1A), 5-HT(1B) and GABA(A) receptors. AB - RATIONALE: Young rodents emit ultrasonic vocalizations (USVs) when separated from their dams and littermates. Pharmacological agents that act on GABA(A) and/or 5 HT receptors and that alleviate anxiety in humans reduce the emission of these calls. OBJECTIVES: 1) to investigate specific 5-HT1 receptor subtypes that modulate maternal separation-induced USVs in mice; 2) to assess the behavioral specificity of these effects; and 3) to compare 5-HT1 agonists with a positive neurosteroid modulator of the GABA(A) receptor complex. METHODS: Seven-day old CFW mouse pups were isolated from their littermates and placed onto a 20 degrees C surface for 4 min. USVs between 30 and 80 kHz, grid crossing, and rectal temperature were measured in separate groups of mouse pups following subcutaneous administration of 5-HT1A and 5-HT1B receptor agonists and antagonists, the neurosteroid allopregnanolone, or the benzodiazepine midazolam. RESULTS: The 5 HT1A agonists (+)8-OH-DPAT (0.01-0.1 mg/kg) and flesinoxan (0.3-1.0 mg/kg), the selective 5-HT1B agonist CP-94,253 (0.03-30.0 mg/kg), and the mixed 5-HT1B/2C receptor agonist TFMPP (0.1-10.0 mg/kg) dose-dependently reduced USVs. These effects were reversed by the 5-HT1A receptor antagonist WAY 100,635 (0.1 mg/kg) or the 5-HT1B/D receptor antagonist GR 127935 (0.1 mg/kg). The effects of TFMPP were biphasic; low doses (i.e. 0.01 and 0.03 mg/kg) increased the rate of vocalization. Midazolam and allopregnanolone also reduced USVs. The highest doses of flesinoxan, (+)8-OH-DPAT, and allopregnanolone suppressed locomotion, whereas CP-94,253, TFMPP, and midazolam stimulated motor activity. CONCLUSIONS: These experiments confirm that agonists at the 5-HT1 receptors and a positive allosteric modulator of the GABA(A) receptor complex decrease maternal separation induced USVs in mice, with 5-HT1B manipulations dissociating the effects on vocalizations from sedative effects. PMID- 10823410 TI - Sucrose intake predicts rate of acquisition of cocaine self-administration. AB - RATIONALE: A number of studies have indicated a relationship between the intake of palatable foods or fluids and drug self-administration. OBJECTIVES: Two experiments were conducted to determine whether the intake of sucrose or fat was related to subsequent cocaine self-administration. METHODS: In separate groups of rats, sucrose or fat was presented for 60 min daily for 7 days. On day 8, a mild stressor (saline injection) was given just prior to sucrose or fat presentation. Rats were then catheterized and tested for cocaine self-administration on a fixed ratio schedule at doses from 0.2 mg/kg to 1.0 mg/kg per infusion and on a progressive ratio schedule at doses from 0.2 mg/kg to 1.5 mg/kg per infusion. RESULTS: Sucrose intake after a mild stressor was significantly related to time to acquisition, with those rats consuming the most sucrose meeting the acquisition criterion sooner than those rats consuming lower amounts of sucrose. Subsequent to acquisition, however, low and high sucrose feeders did not consistently differ in the amount of cocaine self-administered. No relationship was observed between fat intake and rate of acquisition. CONCLUSION: These results provide additional evidence of a relationship between sucrose intake and drug reward, and suggest that stress reactivity may be an important component of this relationship. PMID- 10823411 TI - Characterisation of the effects of nicotine in the five-choice serial reaction time task in rats: antagonist studies. AB - RATIONALE: Nicotine has been shown to decrease reaction time and increase anticipatory responses in a five-choice serial reaction time task (5-CSRTT) in rats, but the receptor mechanisms mediating this effect remain unknown. OBJECTIVES: To evaluate further the effects of nicotine in this task and to characterise the receptors mediating these effects. METHODS: Using a standard 5 CSRTT protocol, rats were trained to respond to a 0.5-s visual stimulus, which was reduced to 0.25 s for experimental sessions to induce a performance decrement. The effects of acute (0.03-0.3 mg/kg IP) and repeated (0.1 and 0.3 mg/kg IP for 5 days) nicotine were studied, as was the ability of mecamylamine (1 mg/kg IP), hexamethonium (5 mg/kg IP), dihydro-beta-erythroidine (6 mg/kg IP) and methyllycaconitine (10 mg/kg IP) to antagonise the effects of acute nicotine. RESULTS: Nicotine had no effect on accuracy, but decreased response latencies, improved performance in the less-well attended stimulus locations and increased inappropriate responding after both acute and repeated treatment. The data suggest that nicotine improves readiness to respond and improves target scanning, and decreases the ability to withhold premature responses (i.e. increased impulsivity). Except for the reduction in error latency, all of the effects of nicotine were antagonised by the non-selective, centrally acting antagonist mecamylamine, whereas the peripheral antagonist hexamethonium had no effect, demonstrating that nicotine's actions are central in origin. Dihydro-beta erythroidine, a competitive nicotinic antagonist, antagonised all of the effects of nicotine. In contrast, the alpha7 antagonist methyllycaconitine had no significant effects against nicotine. CONCLUSIONS: These results demonstrate that the alpha7 receptor subtype is not involved in the effects of nicotine in the 5 CSRTT and that its effects are more likely to be mediated by a receptor(s) such as alpha4beta2, alpha4beta4 and/or alpha3beta2 which is sensitive to antagonism by dihydro-beta-erythroidine. PMID- 10823412 TI - Serotonin releasers increase prepulse inhibition in serotonin 1B knockout mice. AB - RATIONALE: Prepulse inhibition (PPI) is the normal reduction of the startle response which occurs when an abrupt startling stimulus is preceded by a weak pre stimulus and is decreased in several neuropsychiatric disorders. OBJECTIVE: The role of the serotonin 1B (5-HT(1B)) receptor in modulating PPI was investigated using 5-HT-releasing agents in wild-type (WT) and 5-HT1B knockout (1BKO) mice. Whether the differential effects of 5-HT-releasing agents on PPI in WT and 1BKO mice resulted from lack of the 5-HT1B receptor or altered development was also assessed. METHODS: PPI was assessed in WT and 1BKO mice treated with the 5-HT releasing agents (+)3,4-methylenedioxy-N-methylamphetamine (MDMA: 0, 10 mg/kg) or (+/-)N-methyl-1-(1,3-benzodioxol-5-yl)-2-butanamine (MBDB: 0, 10 mg/kg). Additionally, intact 129 Sv mice received pre-treatments of the 5-HT1B/1D antagonist GR 127935 (0, 0.75, 1.5, 3.0 mg/kg) and treatments of MDMA (10 mg/kg). RESULTS: MDMA and MBDB increased PPI in 1BKO mice, but did not alter PPI in WT mice. Intact 129 Sv mice receiving 3.0 mg/kg GR 127935 and 10 mg/kg MDMA exhibited increases in PPI. CONCLUSIONS: The ability of GR 127935 to increase PPI in intact MDMA-treated mice suggests that lack of the 5-HT1B receptor, and not altered development, is responsible for the PPI-increasing effects of 5-HT releasers in 1BKO mice. 5-HT release activates multiple 5-HT receptor subtypes, which individually may increase or decrease PPI and together have a combined effect on PPI. Our finding that MDMA and MBDB increase PPI in 1BKO, but not WT mice, indicates that the activation of 5-HT1B receptors by 5-HT disrupts PPI. PMID- 10823413 TI - Effect of central 5-hydroxytryptamine depletion on inter-temporal choice: a quantitative analysis. AB - RATIONALE: It has been proposed that the ascending 5-hydroxytryptaminergic (5 HTergic) pathways are involved in "impulse control". Previous experiments have shown that rats whose 5-HTergic pathways have been destroyed are more liable than intact rats to select a smaller, immediate reinforcer rather than a larger, delayed reinforcer (impulsive choice). However, it remains unclear whether this effect of central 5-HT depletion reflects a change in the rate of time discounting (i.e. a change in the rate at which reinforcers become devalued as a function of delay) or a change in sensitivity to reinforcer size. OBJECTIVE: We examined the effect of central 5-HT depletion on time discounting using a quantitative model of inter-temporal choice (multiplicative hyperbolic model), which enables effects on time discounting to be differentiated from effects on sensitivity to reinforcer size. METHODS: Rats received injections of 5,7 dihydroxytryptamine into the dorsal and median raphe nuclei or sham lesions. They were trained to press two levers for food-pellet reinforcers in a discrete-trials adjusting-delay schedule. In free-choice trials, selection of lever A resulted in a brief fixed delay (dA) followed by delivery of one pellet; selection of lever B resulted in a longer variable delay (dB) followed by delivery of two pellets; dB was adjusted in accordance with the subject's choices. The value of dA was varied (0.5-8.0 s) in successive phases of the experiment, and the indifference value of dB was determined in each case. RESULTS: In both groups, the indifference value of dB was linearly related to the value of dA, in accordance with the multiplicative hyperbolic model. The lesioned group showed shorter indifference delays than the sham-lesioned group, this being reflected in a parallel displacement of the linear indifference function. In both experiments, the levels of 5-HT and 5-hydroxyindole-acetic acid were reduced in the brains of the lesioned rats, but the levels of noradrenaline and dopamine were not altered. CONCLUSIONS: According to the multiplicative hyperbolic model, parallel displacement of the linear indifference function uniquely specifies a change in time discounting. Thus these results indicate that central 5-HT depletion results in an increase in the rate of time discounting for food reinforcers. PMID- 10823414 TI - Selective impairments in the stress response in schizophrenic patients. AB - OBJECTIVE: In the vulnerability-stress concept of schizophrenia, schizophrenic patients are thought to display increased sensitivity to stress. Little is known about the biological mechanisms that are involved in stress processing in schizophrenic patients. In this study, hypothalarnic-pituitary-adrenal (HPA) function in schizophrenic patients was studied for its essential role in stress processing and adaptation to the environment. METHODS: Eighteen schizophrenic patients were compared to 21 healthy controls in their salivary cortisol response to a physical (bicycle ergometry) and a psychosocial (public speaking) stressor. Coping questionnaires were included as a measure of stress processing at the psychological level. Basal HPA function was assessed by measuring cortisol day profiles and feedback activity by using dexamethasone and hydrocortisone. RESULTS: Schizophrenic patients showed blunted cortisol responses to the psychosocial stressor, but not to the physical stressor, in spite of similar increases in heart rate. The cortisol response to the psychosocial stressor tended to be negatively correlated to the use of passive and avoidant coping strategies. Basal HPA function appeared intact in the schizophrenic patients. CONCLUSIONS: The findings show a selective impairment in the response to psychosocial stress in schizophrenic patients. This suggests the involvement of brain systems that play a role in the activation of the HPA system to psycho social stress, like arginin-vasopressin (AVP), and cognitive processes, like coping. PMID- 10823415 TI - Macrophage infiltration in human non-small-cell lung cancer: the role of CC chemokines. AB - Bronchogenic carcinoma is the leading cause of malignancy-related mortality in the United States, with an overall 5-year survival rate of less than 15%. This aggressive behavior reflects, among other traits, the capacity of the tumor to evade normal host immune defenses, and to induce a pro-angiogenic environment. A central feature of any immune response toward tumors is the recruitment of specific immune cell populations. In the present study we investigated the infiltration of monocytes in human specimens of non-small-cell lung cancer (NSCLC). The presence of macrophages in NSCLC tumors was documented by immunohistochemistry. In vitro chemotaxis assays demonstrated higher monocyte chemotactic activity in NSCLC tumor homogenates than in normal lung tissue. We next investigated the expression of CC chemokines within specimens of NSCLC tumors. Levels of the CC chemokines were higher in NSCLC tumor tissue than in normal lung tissue. Immunolocalization showed that the cells associated with antigenic CC chemokines were the malignant tumor cells, as well as occasional stromal cells. Maximal inhibition of monocyte chemotaxis induced by NSCLC in vitro occurred in the presence of neutralizing antibodies to MCP-1 and MIP-1beta. On follow-up of 15 patients in whom we quantified macrophage infiltration, we found that those with recurrence of disease had higher levels of macrophage infiltration in their initial tumors. However, the functional significance of CC chemokine-mediated macrophage infiltration into NSCLC remains to be determined. PMID- 10823416 TI - Synergistic effect of interleukin-15 and interleukin-12 on antitumor activity in a murine malignant pleurisy model. AB - Interleukin(IL)-15, which uses IL-2 receptor (R) beta and gamma chains for signal transduction, shares many of the biological activities of IL-2. We examined the effects of exogenous IL-15 on protection in a murine malignant pleurisy model using BALB/c mice and syngeneic MethA fibrosarcoma (MethA). Intrapleural administration of IL-15 significantly prolonged the survival time of mice after an intrapleural inoculation of MethA, whereas the same dose of IL-2 did not. The in vivo antitumor effect of IL-15 was synergistically enhanced by additive administration of IL-12. Combination therapy of IL-15 and IL-12 protected mice from death from bloody pleural fluid. Such treatment induced marked increases in the number of CD3-IL-2Rbeta+ cells corresponding to natural killer (NK) cells and the production of interferon gamma (IFNgamma) by T cells in the thoracic exudate cells (TEC). Administration of anti-IFNgamma mAb partly inhibited the protective effect of a combination of IL-15 and IL-12. A tumor-neutralizing (Winn) assay revealed that the antitumor activity of effector cells in the TEC was abrogated by treatment with anti-CD8 mAb or anti-asialoGM1 Ab plus complement. Thus, treatment with IL-15 in combination with IL-12 may enhance the activities of NK and CD8+ T cells in the TEC, providing strong antitumor activity against the malignant pleurisy. These results suggest that IL-15 together with IL-12 may have potential for the immunotherapy of some types of malignant pleurisy. PMID- 10823417 TI - Paclitaxel up-regulates interleukin-8 synthesis in human lung carcinoma through an NF-kappaB- and AP-1-dependent mechanism. AB - Lung cancer is a leading cause of cancer-related death in the United States. For this reason we chose to study the specific cellular effects that one chemotherapeutic agent, paclitaxel, has on lung carcinoma. In addition to its known mechanism of action, which is to stabilize microtubules, paclitaxel has been shown to have other interesting and relevant cellular effects. In this report, we demonstrate that a subset of human lung carcinoma cell lines respond to paclitaxel treatment with an up to a fivefold increase in the production of interleukin-8 (IL-8). We demonstrate that this increased production is specific to IL-8 but not to other chemokines, and is both dose- and time-dependent. Increased IL-8 mRNA is seen as early as 45 min with a peak at 4 h after paclitaxel treatment. This increase in mRNA is due to transcriptional activation because actinomycin D treatment blocked the increase. Paclitaxel also activates the mitogen-activated protein kinase family member, JNK1, in dose-dependent fashion. IL-8 enhancement is completely abolished with the use of an inhibitor of NF-kappaB, the super-repressor IkappaB. Similar results were obtained upon the inhibition of AP-1 activation with the MEK1/2 inhibitor, U0126. By gaining a better understanding of the differences in cellular response to paclitaxel chemotherapy, these findings might lead to either improved patient selection or to the development of adjuvant therapy targeted at specific-cell signaling proteins. PMID- 10823418 TI - Antigenicity of human melanoma cells transfected to express the B7-1 co stimulatory molecule (CD80) varies with the level of B7-1 expression. AB - The aim of this study was to compare the antigenicity of human melanoma cells molecularly modified by particle-mediated gene transfer to have transient or stable expression of the B7-1 co-stimulatory molecule (CD80). The unmodified melanoma cells (mel5, m21) had no constitutive expression of B7-1, but 22%-28% of cells had transient B7-1 expression 24 h following transfection with cDNA for B7 1 (mel5-B7, m21-B7). In addition, 85%-90% of cells had stable B7-1 expression following transfection with cDNA for B7-1 and in vitro culture under selection conditions (mel5-B7neo, m21-B7neo). Allogeneic HLA-unmatched normal donor peripheral blood mononuclear cells (PBMC) secreted greater amounts of granulocyte/macrophage-colony-stimulating factor (GM-CSF) when incubated for 3 days with m21-B7neo than did PBMC incubated with m21-B7, which, in turn, secreted greater amount of GM-CSF than PBMC incubated with m21. Similarly, cell-mediated cytotoxicity against unmodified melanoma cells by PBMC co-cultured for 5 days with the modified or unmodified melanoma cells was proportional to the level of B7-1 expression on the stimulating cells. This cytolytic activity had both an HLA class-I-restricted and an HLA-class-I-unrestricted component. Following 5 days of co-culture, PBMC expression of CD28, the ligand for B7-1, was down-regulated in proportion to the level of B7-1 expression on the stimulating melanoma cells. Thus, particle-mediated gene delivery of cDNA for B7-1 into human melanoma cells increased expression of functional B7-1 and enhanced the antigenicity of the gene modified cells in proportion to their level of B7-1 expression. PMID- 10823419 TI - Effects of macrophage-colony-stimulating factor on cyclophosphamide-injected mouse NK1.1+ cell activity. AB - We injected cyclophosphamide into mice and examined their natural killer (NK) activity both in vitro and in vivo. Cyclophosphamide injection temporarily abrogated the lung clearance activity of Yac-1 lymphoma cells, which is considered to be an index of NK activity in vivo. However, administration of recombinant human macrophage-colony-stimulating-factor (rhM-CSF) to cyclophosphamide-injected mice restored the lung clearance activity. To clarify whether the administration of rhM-CSF activated NK cells, we purified NK1.1+ cells from mice treated with cyclophosphamide and/or rhM-CSF and examined their functions (cytotoxicity, proliferation, and interferon gamma production) in vitro. Cyclophosphamide injection decreased the number, but did not suppress the functions of NK1.1+ cells. The numbers of NK1.1+ cells in cyclophosphamide injected mice restored by rhM-CSF administration. And the functions of NK1.1+ cells from both saline-injected and cyclophosphamide-injected mice were accelerated by rhM-CSF administration. These results suggested that the temporary abrogation of NK activity in vivo caused by cyclophosphamide injection was due to a decrease in the number and not to suppression of the functions of NK1.1+ cells. The injection of cyclophosphamide into mice increased the number of tumor (B16 melanoma) nodules formed in the lungs and liver. However, treatment with rhM-CSF recovered the anti-metastatic activity in the lungs of cyclophosphamide-injected mice. These results show that administration of rhM-CSF restores NK activity suppressed by cyclophosphamide injection in vivo. PMID- 10823420 TI - CD40 expression on human lung cancer correlates with metastatic spread. AB - PURPOSE: The poor prognosis associated with lung cancer is related to the high incidence of regional and distant metastasis. There is a crucial need to identify parameters that can predict a tendancy to metastatic spread to allow better prognostic evaluation and therapeutic approach. METHODS: Using flow cytometry we evaluated 18 human lung cancer cell lines for the expression of different surface markers on lung cancers suggested to be possible prognostic parameters, including epidermal growth factor receptor (EGFR), intercellular adhesion molecule 1 (ICAM 1), Fas and CD40. RESULTS: No correlation was found between tumor prognosis and EGFR, ICAM-1 or Fas. However, a statistically significant correlation was found between the surface expression of CD40 and the metastatic spread of the tumor. In this study, 14 of 18 lung cancer cell lines (78%) expressed CD40 on their surface. All of the 4 tumors that were CD40-negative, were stage I tumors, without any evidence of regional or distant metastasis. Of the 14 tumors that expressed CD40, all but 1 (93%) had either nodal or systemic metastasis at the time of diagnosis. Patients whose tumors were CD40-negative showed a significantly better N stage, overall stage at presentation and survival than those patients with CD40-positive patients. No significant differences between the two groups were observed in tumor size, gender, age, histology, differentiation or preoperative therapy. CONCLUSIONS: These results suggest that CD40 expression on lung cancer may play a role in metastatic spread, and also may serve as a prognostic marker and an indicator of advanced disease. PMID- 10823421 TI - Target-cell-induced anergy in natural killer cells: suppression of cytotoxic function. AB - Our earlier studies have demonstrated that natural killer (NK) cells are the effectors that participate during the spontaneous regression of AK-5 tumour in syngeneic hosts. We have shown that the tumour cells are killed by necrosis and apoptosis. In this study, we have examined the induction of functional anergy in NK cells following coculture with fixed AK-5 tumour cells at high ratio. NK cells, upon coculture with fixed AK-5 cells (1:1 ratio), showed loss of cytotoxic function against both AK-5 (antibody-dependent cell cytotoxicity) as well as YAC 1 targets. The response of these cells to the activation by recombinant interleukin-2 and recombinant interferon gamma was poor. Induction of tumour necrosis factor alpha (TNFalpha) secretion was observed after coculture of NK cells with fixed AK-5 cells. The cocultured cell supernatant inhibited the cytotoxic activity of NK cells, which was partially restored with anti-TNFalpha antibody. In addition, NK cells, after treatment with fixed tumour cells showed overexpression of the Fas receptor. We have also observed induction of apoptosis in cocultured NK cells. These studies suggest that the fixed tumour cells (antigen) at high ratio are able to suppress NK cell function as well as induce death in NK cells. PMID- 10823422 TI - Local cytokine therapy of cancer: interleukin-2, interferons and related cytokines. AB - Local therapy with interleukin-2 (IL-2) and other cytokines may be a very effective way to treat cancer. This was the theme of the First Symposium on Local Cytokine Therapy of Cancer: Interleukin-2, Interferons and Related Cytokines, in Hamburg, 29 April-1 May 1999. The abstracts are published in Anticancer Research 19: 1995-2016 (1999). Here we present a report. PMID- 10823423 TI - The vertebral end-plate: what do we know? AB - The vertebral end-plate is critical for maintaining the health of the intervertebral disc. This paper reviews the literature that relates to end-plate structure and development and describes how disc integrity is affected by changes associated with pathology and aging. A range of studies, including biomechanical, biochemical, surgical and histological is discussed to illustrate the important role of the end-plate in spinal function. PMID- 10823424 TI - Titanium-alloy enhances bone-pedicle screw fixation: mechanical and histomorphometrical results of titanium-alloy versus stainless steel. AB - Several types of pedicle screw systems have been utilized to augment lumbar spine fusion. The majority of these systems are made of stainless steel (Ss), but titanium-alloy (Ti-alloy) devices have recently been available on the market. Ti alloy implants have several potential advantages over Ss ones. High bioactivity and more flexibility may improve bone ingrowth and mechanical fixation, and the material also offers superior magnetic resonance imaging (MRI) and computed tomography (CT) resolution and significantly less signal interference. However, no data are available from loaded spinal constructs regarding bony ingrowth and mechanical fixation. The aim of this study was to analyse the effect of Ti-alloy versus Ss pedicle screws on mechanical fixation and bone ingrowth in a loaded mini-pig model. Eighteen adult mini-pigs underwent total laminectomy and posterolateral spinal fusion at L3-L4, and were randomly selected to receive either Ss (n = 9) or Ti (n = 9) pedicle screw devices. In both groups, the device used was compact Cotrel-Dubousset instrumentation (Sofamore Danek) of an identical size and shape. The postoperative observation time was 3 months. Screws from L3 were used for histomorphometric studies. Mechanical testing (torsional tests and pull-out tests) was performed on the screws from L4. The Ti screws had a higher maximum torque (P < 0.05) and angular stiffness (P < 0.07), measured by torsional testing. In the pull-out tests, no differences were found between the two groups with respect to the maximum load, stiffness and energy to failure. No correlation between removal torque and the pull-out strength was found (r = 0.1). Bone ongrowth on Ti was increased by 33% compared with Ss (P < 0.04), whereas no differences in bone volume around the screws were shown. Mechanical binding at the bone-screw interface was significantly greater for Ti pedicle screws than for Ss, which was explained by the fact that Ti screws had a superior bone ongrowth. There was no correlation between the screw removal torque and the pull-out strength, which indicates that the peripheral bone structure around the screw was unaffected by the choice of metal. PMID- 10823425 TI - Load-displacement properties of the normal and injured lower cervical spine in vitro. AB - The objective of this study was to determine which discoligamentous structures of the lower cervical spine provide significant stability with regard to different loading conditions. Accordingly, the load-displacement properties of the normal and injured lower cervical spine were tested in vitro. Four artificially created stages of increasing discoligamentous instability of the segment C5/6 were compared to the normal C5/6 segment. Six fresh human cadaver spine segments C4-C7 were tested in flexion/extension, axial rotation, and lateral bending using pure moments of +/- 2.5 Nm without axial preload. Five conditions were investigated consecutively: (1) the intact functional spinal unit (FSU) C5/6; (2) the FSU C5/6 with the anterior longitudinal ligament and the intertransverse ligaments sectioned; (3) the FSU C5/6 with an additional 10-mm-deep incision of the anterior half of the anulus fibrosus and the disc; (4) the FSU C5/6 with additionally sectioned ligamenta flava as well as interspinous and supraspinous ligaments; (5) the FSU C5/6 with additional capsulotomy of the facet joints. In flexion/extension, significant differences were observed concerning range of motion (ROM) and neutral zone (NZ) for all four stages of instability compared to the intact FSU. In axial rotation, only the stage 4 instability showed a significantly increased ROM and NZ compared to the intact FSU. For lateral bending, no significant differences were observed. Based on these data, we conclude that flexion/extension is the most sensitive load-direction for the tested discoligamentous instabilities. PMID- 10823426 TI - Whiplash-type neck distortion in restrained car drivers: frequency, causes and long-term results. AB - An analysis was made of 1176 whiplash-type neck distortions taken from a total of 3838 restrained car driver incident reports. The percentage of whiplash-type neck distortion among injured drivers increased from less than 10% in 1985 to over 30% in 1997. Most occurred in head-on crashes or crashes with multiple collisions; only 15% occurred in rear-end collisions. More than 1,000 questionnaires were sent to the injured to find out about the duration and type of complaints caused by their cervical spine injury. Although only 138 (12%) returned the questionnaire, which may not be a representative sample, a further analysis was carried out. Of the 138, 121 (88%) indicated that they had suffered or were still suffering from their symptoms. The percentages of the various complaints were as follows: pain (74%), tension (6%) and stiffness (5%) in the head (27%), neck (55%) and shoulder (8%). The duration of the complaints was longest after multiple collisions and when the onset of complaints was longer than 24 h after trauma. Women and elderly persons predominated slightly in the group with longer duration of complaints. A correlation between the severity of the accompanying injuries and duration of complaints was found. Lack of adequate follow-up for patients with less severe injuries posed considerable difficulties for this retrospective study. In order to better evaluate this problem, prospective studies are necessary, with documentation including diagnosis, treatments, complaint duration and type. PMID- 10823427 TI - Surgical treatment of nonunited fractures of the odontoid process, with special reference to occipitocervical fusion for unreducible atlantoaxial subluxation or instability. AB - Fifty-seven consecutive patients treated surgically for nonunited fractures of the odontoid process were reviewed. All patients presented late, exhibiting neurological deficits subsequent to nonunion. Delay in presentation was between 6 and 120 months (mean 32 months) after the original injury, due to missed diagnosis or inappropriate management. Seven patients who were reduced in traction underwent a Gallie atlantoaxial fusion. In the remaining 50 patients who were unreducible, an occipitocervical arthrodesis was performed. They were followed up for a minimum of 2 years, except one who died from postoperative respiratory failure. All patients obtained a solid bony union, including two in whom nonunion occurred following atlantoaxial fusion, and occipitocervical fusion was added as a rescue. Thirty-eight patients achieved excellent neurological recovery, nine still had some disability, five retained their neurological deficits and two reported a deterioration. In two patients, a recurrence in a traumatic episode was experienced long after a resolution. Our findings demonstrate that occipitocervical arthrodesis is preferable for unreducible subluxation or instability of atlantoaxial articulation in nonunion of odontoid fractures. PMID- 10823428 TI - Traumatic spondylolisthesis of the axis: treatment rationale based on the stability of the different fracture types. AB - Thirty-nine consecutive patients, 22 male and 17 female with an average age of 37.6 years, with traumatic spondylolisthesis of the axis were reviewed. The cause of injury in 75% of the patients was a road traffic accident. The fractures were classified according to Effendi et al., the type II fractures were further divided into three subgroups: flexion, extension and listhesis injuries. There were 10 type I (25.7%) and 29 type II fractures (74.4%); of these, 12 (30.8%) were classified as flexion-type, 2 (5.1%) as extension-type and 15 (38.5%) as listhesis-type. We did not identify any case of type III injury. Overall, 43.5% of the patients had sustained a significant head or chest trauma, with the highest incidence for type II listhesis injuries. Significant neurological deficits occurred in four patients (10.3%); in all four,the fracture was classified as a type II listhesis. All ten type I injuries were successfully treated with a cervical orthosis. Ten of the 12 type II flexion injuries demonstrated significant angulation. Two were treated with internal stabilisation, in seven with a halo device and one with a minerva plaster of Paris (PoP). Healing was uneventful in all ten patients. For the remaining two stable type II flexion injuries, application of a hard collar was adequate, as was the case for the two stable type II extension injuries. Six of the 15 type II spondylolisthesis injuries underwent primary internal stabilisation, and healing was uneventful in all cases. In four (44.4%) of the nine injuries that were primarily treated with a halo device/minerva PoP, secondary operative stabilisation had to be performed. The classification of Effendi et al. provides a complete description of the different fractures. However, further distinction of the type II injuries regarding their stability is mandatory. Type II spondylolisthesis injuries are unstable, with a high number of associated injuries, a great potential for neurological compromise and significant complications associated with non-operative treatment. The majority of type II extension and type II flexion injuries can be successfully treated with nonrigid external immobilisation. PMID- 10823429 TI - A systematic literature review to identify the best method for a single level anterior cervical interbody fusion. AB - The goal of this systematic literature review was to determine, for patients with degenerative disc disease, which method of single-level anterior cervical interbody fusion using the anterior approach gives the best clinical and radiological outcome. The number of new techniques for obtaining a solid fusion has increased rapidly, but the rationale for choosing between different techniques is unclear. Randomised comparative studies on anterior cervical interbody fusions were identified in a sensitive Medline, Cochrane and Current Contents database search. Two independent reviewers evaluated the articles that met the selection criteria, using a checklist. The search yielded eight randomised, controlled trials for the systematic literature review. Three of these studies were judged to be of sufficient quality with regard to methodology and the information provided. In the three articles, five different treatment methods were investigated, four of which were interbody fusions. Fusion rates varied between 28% for an allograft method and 63% for a discectomy-alone method. In one study, kyphosis varied from 40% to 62% between treatments. Good clinical outcome (disability, pain and symptoms) ratings varied from 66% to 82%. A meta analysis to determine the best method for an anterior interbody fusion could not be performed due to the heterogeneity of the methods reported and because no standard outcome parameter was used. From this systematic literature review, a gold standard for the treatment of degenerative disc disease could not be identified. PMID- 10823430 TI - Outcome scores in degenerative cervical disc surgery. AB - Forty-six consecutive patients with neck pain and arm radiculopathy were treated with anterior cervical discectomy and fusion. All patients had neurological symptoms corresponding to a herniated disc and/or spondylosis at one or two cervical levels, verified by magnetic resonance imaging. The patients were stabilized with an anterior graft and randomized to either fixation with a CSLP plate or no internal fixation. Preoperatively and 2 years postoperatively the patients filled in a questionnaire that included a modified Million Index, a modified Oswestry Index and the Zung Depression Scale. They were also asked to register their pain in the arm and in the neck on a vertical visual analogue scale (VAS). At the 2-year follow-up, an unbiased observer graded the patients' clinical outcome using Odom's criteria. A test-retest procedure was carried out to examine the questionnaire reproducibility. In the group that was operated at one level, there was no significant improvement in any of the scores. Nevertheless, 81% of the patients were satisfied with the outcome of the surgery. All scores improved in the group operated at two levels. The pain in the neck and arm, as measured on a VAS, decreased in both groups. The improvement in arm pain was significantly more pronounced in patients operated with a plate at two levels compared to those who were operated without a plate. At the 2-year follow-up, patients with an excellent or good result according to Odom's criteria had a lower Million Index (P < 0.0005), Oswestry Index (P < 0.0005), and Zung (P = 0.024) score, than the group classified as fair or poor. There was a significant correlation (P < 0.0001 for all scores) between the test and retest results. We conclude that the modified Million Index and Oswestry Index are clinically useful tools in the evaluation of outcome after degenerative cervical disc surgery. The clinical benefits of plate fixation were minimal. The outcome after surgery, measured with the Oswestry Index, Million Index and VAS for arm and neck pain, seems to correlate well with the classification of outcome by Odom. PMID- 10823431 TI - Boomerang deformity of cervical spinal cord migrating between split laminae after laminoplasty. AB - Patients with cervical compression myelopathy were studied to elucidate the mechanism underlying boomerang deformity, which results from the migration of the cervical spinal cord between split laminae after laminoplasty with median splitting of the spinous processes (boomerang sign). Thirty-nine cases, comprising 25 patients with cervical spondylotic myelopathy, 8 patients with ossification of the posterior longitudinal ligament, and 6 patients with cervical disc herniation with developmental canal stenosis, were examined. The clinical and radiological findings were retrospectively compared between patients with (B group, 8 cases) and without (C group, 31 cases) boomerang sign. Moderate increase of the grade of this deformity resulted in no clinical recovery, although there was no difference in clinical recovery between the two groups. Most boomerang signs developed at the C4/5 and/or C5/6 level, where maximal posterior movement of the spinal cord was achieved. Widths between lateral hinges and between split laminae in the B group were smaller than in the C group. Flatness of the spinal cord in the B group was more severe than in the C group. In conclusion, the boomerang sign was caused by posterior movement of the spinal cord, narrower enlargement of the spinal canal and flatness of the spinal cord. PMID- 10823432 TI - Hartshill rectangle: failure of spinal stabilisation in acute spinal cord injury. AB - A high rate of failure of the internal fixation of unstable spinal fractures in complete cord injured patients was noted in patients referred to the Salisbury Spinal Centre who had been stabilised with a Hartshill rectangle. This prompted a review of the operative notes, radiographs and clinical outcomes of all patients referred to the centre with a Hartshill rectangle in situ. All patients identified with a complete spinal cord injury and Hartshill rectangle were identified. Forty-three such patients referred from 13 different centres were found. Pre- and postoperative radiographs were assessed for fracture pattern and for spinal correction. Operative outcome in terms of pain and complications relating to surgery were identified. The most recent radiographs were assessed for signs of loss of reduction or stabilisation. Follow-up averaged 84 months (range 36-132 months). Of the 43 identified patients, 19 were found to have unsatisfactory stabilisation. Persistent pain, broken implants and worsening kyphosis were the main complications. The failure to use bone graft at the time of stabilisation was significantly (P < 0.001) related to risk of failure. The application and use of the Hartshill is not a technically challenging procedure; however, if the system is to be used, it must be used correctly. Failure to correctly apply the rectangle and to use bone graft will lead to an unacceptably high rate of failure. PMID- 10823433 TI - Decompensation following scoliosis surgery: treatment by decreasing the correction of the main thoracic curve or "letting the spine go". AB - Coronal decompensation following correction of adolescent idiopathic scoliosis (AIS) has been reported to be due to the Cotrel-Dubousset rod derotation maneuver, or to a hypercorrection of the main thoracic curve. The treatment of such decompensation consists classically in observation, bracing, or extension of the instrumentation in the lumbar spine for a King 2 curve, or in the upper thoracic spine for a King 5 curve. As the postoperative decompensation is related to a hypercorrection of the main thoracic curve (relative to the compensatory curve), we hypothesized that if we were to "let the spine go" to some of its initial deformity, the balance of the patient would be improved. The purpose of the study was therefore to report on two cases where a postoperative imbalance following scoliosis surgery was successfully treated by decreasing the correction of the main thoracic curve. Two patients with AIS were found to have significant imbalance after scoliosis surgery. Both patients had been treated for a right thoracic curve (82 degrees and 85 degrees respectively) with an anterior release and posterior instrumentation. The revision surgery consisted for both patients in removing all the hooks between the end vertebrae of the main thoracic curve. This was done before the 3rd postoperative month for both patients. After revision surgery, the balance of both patients improved dramatically within a few weeks. The shoulders became almost level, and the trunk shift improved concomitantly. The Cobb angle increased by 8 degrees and 10 degrees, and the apical vertebra shifted to the right by 15 and 10 mm for the respective patients. These results were stable at 1-year follow-up. In the event of a persisting imbalance, we recommend, in selected cases, letting the spine go by removing all the implants located between the end vertebrae of the main thoracic curve. This adjustment or fine-tuning of the instrumentation should be done before the fusion takes place, and is best achieved with an instrumentation in which the hooks can be easily removed from the rod. PMID- 10823434 TI - Evaluation of clinical tests used in classification procedures in pregnancy related pelvic joint pain. AB - Pain in the pelvic joints and lower back, a major problem for pregnant women, has proved resistant to precise measurement and quantification. To develop a classification system, the clinical tests used must be able to separate pelvic from low back pain; they must also have a high inter-examiner reliability, sensitivity and specificity, and preferably be easy to perform. The aim of this study was to describe a standardised way of performing tests for examining the pelvis, and to evaluate inter-examiner reliability, and establish the sensitivity and specificity of 15 clinical tests. It was designed as a longitudinal, prospective, epidemiological cohort study. First, 34 pregnant women were examined by blinded examiners to establish inter-examiner reliability. Second, a cohort of 2269 consecutive pregnant women, each responded to a questionnaire and underwent a thorough and highly standardised physical examination (15 tests with 48 possible responses) of the pelvic joints and surrounding areas. The 535 women who reported daily pain from the pelvic joints and had objective findings from the joints were divided, according to symptoms, into four classification groups and one miscellaneous group. The results of the study showed inter-examiner agreement of the tests was high, calculated in percentage terms, at between 88 and 100%. Using the Kappa coefficient, most tests kept the high agreement: six tests had an inter-examiner agreement of between 0.81 and 1.00, three between 0.61 and 0.80, and two between 0.60 and 0.41. Five tests showed superior sensitivity. The specificity of the tests was between 0.98 and 1.00, except the value for pelvic topography, which was 0.79. These results show that it is possible to standardise examination and interpretation of clinical tests of the pelvic joints, resulting in a high degree of sensitivity, specificity and inter-examiner reliability. PMID- 10823435 TI - Thoracolumbar distraction fractures in advanced pregnancy: a contribution of two case reports. AB - Thoracolumbar trauma in pregnant women is an important topic, though rarely discussed in the pertinent literature. Two consecutive cases of thoracolumbar distraction injuries in advanced pregnancy are presented. Conservative treatment failed in both cases; surgical management was necessary on a delayed basis using compression instrumentation. The similar features of the pattern of injury in the two patients suggest a cushioning effect of the gravid uterus. A theoretical analysis of the pathomechanics is carried out in an attempt to explain the specificity of flexion-distraction injuries in victims in an advanced stage of pregnancy. The authors suggest that the spine pivots about the fetus, and so is injured more in tension than in compression. The damage is mainly to soft-tissue structures, while anterior column compression and axial loading are less important. PMID- 10823436 TI - Acute pancreatitis complicating anterior lumbar interbody fusion. AB - Postoperative pancreatitis may occur following surgery in regions remote from the pancreas and the biliary tree. Though uncommon, it carries a high mortality rate. Pancreatitis complicating spinal surgery is extremely rare. This report describes a case of acute pancreatitis following an anterior lumbar interbody fusion and discusses the possible mechanisms of pancreatic cellular injury. PMID- 10823437 TI - Evaluation of Crohn disease activity with magnetic resonance imaging. AB - BACKGROUND: The purpose of this study was to assess the accuracy of magnetic resonance imaging (MRI) in evaluating Crohn disease (CD) activity. The intestinal inflammatory activity is usually present in patients under pharmacologic treatment, despite their clinical remission. METHODS: Twenty patients with CD, all under pharmacologic treatment, were prospectively studied by MRI at 1.5 T as a periodic control. Positivity of three acute-phase reactants was considered an index of biologic activity (BA). T2-weighted, T2-weighted fat-suppressed turbo spin-echo, and breath-hold T1-weighted turbo field-echo sequences, before and after gadolinium intravenous injection, were obtained. A negative superparamagnetic contrast agent was orally administered. The following MRI parameters were qualitatively evaluated by three radiologists at the level of the affected bowel and compared with clinical data: wall thickness (WT), wall T2 weighted signal (T2W), wall contrast enhancement (WE), amount of fibrofatty proliferation (FP), and T2-weighted signal of fibrofatty proliferation on fat suppressed images (T2FP). The kappa coefficient of agreement was calculated. The Spearman rank correlation was used for the analysis of clinical and radiologic data. RESULTS: Nineteen of 20 patients were in clinical remission (Crohn Disease Activity Index < 150). On the basis of laboratory tests, nine of 20 patients had biologically active disease. An excellent correlation was found between BA and WE, T2W, and T2FP (0.900, 0.927 and 0.961, respectively; p < 0.0001), and a lower correlation was found between BA and WT and between BA and FP (0.78 and 0.62). Excellent statistical correlation was also found between WE and T2W and between WE and T2FP (0.876 and 0.892). CONCLUSIONS: An excellent statistical correlation was found between biologically "active" disease and the following MRI parameters: wall gadolinium enhancement, wall hyperintensity on T2-weighted fat-suppressed images, and hyperintensity of fibrofatty proliferation on T2-weighted fat suppressed images. Therefore, MRI can be valuable in assessing CD activity. PMID- 10823438 TI - MRI evaluation of Crohn disease activity. PMID- 10823439 TI - Conservative treatment of large rectus sheath hematoma in patients undergoing anticoagulant therapy. AB - BACKGROUND: The purpose of this study was to describe our experience in the conservative management of large rectus sheath hematoma (RSH) in patients undergoing anticoagulation therapy. METHODS: This is a retrospective study of the medical histories of 12 cases of RSH (11 female, one male; mean age = 67.6 years). Seven patients were taking oral anticoagulants, three were taking intravenous unfractionated heparin, and two were taking subcutaneous low molecular-weight heparin. Six patients had a history of coughing fits. Ultrasound examination and computed tomography (CT) was performed in all cases. RESULTS: Clinically, the majority of patients presented acute abdominal pain, infraumbilical masses, and anemic syndrome. Ultrasonography demonstrated nine of the 12 cases of RSH, and CT showed the hematoma in all 12 cases. Type II (five cases) and type III (seven cases) indicate moderate and severe hematomas, respectively. Excessive anticoagulation was observed in eight cases, and coagulation within correct ranges was seen in the remaining four cases. In five patients the normalization of coagulation was achieved by administering vitamin K1 and fresh frozen plasma. All cases of type III hematoma required blood transfusion. Conservative treatment was effective in all cases. CONCLUSIONS: RSH must be suspected in women of advanced age undergoing treatment with anticoagulants who present the clinical triad of acute abdominal pain, infraumbilical mass, and anemic syndrome. CT is the examination of choice for the diagnosis of RSH. Early diagnosis of RSH permits conservative management, even in the case of large hematomas with hemodynamic repercussions and avoids unnecessary surgical intervention. PMID- 10823440 TI - Periesophageal mediastinal fibromatosis. AB - We report two cases of aggressive fibromatosis of the mediastinum with esophageal involvement in adults. CT showed the posterior mediastinal mass involving the esophagus. In both cases, surgical resection was not feasible. Histopathologic examination showed a fibrous tissue. The patients died two years later. PMID- 10823441 TI - Percutaneous gastrostomy in oncologic patients: analysis of results and expansion of the indications. AB - BACKGROUND: Percutaneous gastrostomy is generally performed for permanent enteral nutrition or gastric decompression. METHODS: In our series of oncologic patients, percutaneous gastrostomy was also used temporarily in some patients for enteral nutrition while awaiting functional recovery of swallowing, in preparation for surgery, or for the treatment of fistulas in the upper digestive tract. Fifty-one procedures were performed in 50 patients: 42 for feeding, eight for decompression, and one for transgastric drainage of a duodenal fistula. RESULTS: Of the 35 patients treated for permanent enteral nutrition, four are still alive, with a total survival time of 2,167 days. In three patients, gastrostomy was performed for temporary feeding and was removed once the ability to swallow was restored. In four patients, it was created to restore metabolic balance before surgery. In the patient with a duodenal fistula, healing was achieved in 19 days. The seven patients in whom the procedure was performed for decompression survived for a mean of 19.2 days. There was only one major procedure-related complication (peritonitis). CONCLUSIONS: Percutaneous gastrostomy is a safe, low-cost method that allows the patient to maintain essential nutrition without the discomfort of a nasogastic tube and therefore warrants wider and earlier use. We feel that its application should also be extended to temporary feeding of patients about to undergo long courses of chemotherapy and radiotherapy, which can lead to severe deterioration of nutritional status. PMID- 10823442 TI - Cisapride or metoclopramide to accelerate small bowel transit during barium follow-through examination? AB - BACKGROUND: Metoclopramide is commonly used to accelerate small bowel transit during barium follow-through (BaFT) examinations, but its action is unpredictable. Cisapride, commonly used to treat gastroesophageal reflux disease, also accelerates small bowel transit and may be a viable alternative. The two were compared in a prospective, randomized, blind study. METHODS: Patients attending for BaFT were randomized to receive either 10 mg cisapride or 20 mg metoclopramide orally 1 h before the barium suspension. BaFT was performed by using a standard technique, and small bowel transit and study quality were compared. Patients also noted any side effects experienced. RESULTS: Of 45 patients recruited, 27 received cisapride and 18 metoclopramide. Median transit time for the cisapride group was 30 min (range = 10-130 min) versus 67.5 min (range = 30-290 min) for the metoclopramide group (p = 0.019). Study quality was comparable. However, nine patients (33%) receiving cisapride experienced nausea versus only one subject (6%) receiving metoclopramide (p = 0.034). CONCLUSIONS: This study suggests that cisapride is a more effective prokinetic agent than metoclopramide, but this benefit is offset by a higher incidence of side effects. PMID- 10823443 TI - Needle-tract implantation in hepatocellular carcinoma: frequency and CT findings after biopsy with a 19.5-gauge automated biopsy gun. AB - BACKGROUND: Needle-tract implantation is an important complication of cutting biopsy of hepatocellular carcinoma (HCC). This study was performed to evaluate the frequency of needle-tract implantation after ultrasound (US)-guided percutaneous biopsy of HCC and to describe triple-phase helical computed tomographic (CT) findings of implanted nodules. METHODS: Between April 1994 and December 1997, 205 patients underwent US-guided percutaneous biopsy for HCC. Review of medical records and the pathology database disclosed seven patients who were found to have needle-tract implantation of HCC. Among these patients, five underwent triple-phase helical CT examination. We analyzed the frequency of needle-tract implantation and triple-phase helical CT findings of implanted nodules, with particular attention to the morphology and enhancement pattern. RESULTS: Seven of 205 patients (3.4%) had tumor implantation along the needle tract at histologic examination after surgical resection. Eight implanted nodules in five patients were found on triple-phase helical CT images (one nodule in three patients, two nodules in one patient, and three nodules in one patient). All implanted nodules has well-circumscribed margins and were ovoid or lobulated in contour. On triple-phase helical CT, six (75%) implanted nodules were isodense compared with abdominal wall muscle on all triple-phase CTs, and two (25%) nodules were hyperdense on hepatic arterial and portal venous phases and isodense on equilibrium phase. CONCLUSIONS: The frequency of needle-tract implantation of HCC after percutaneous needle biopsy was higher than reported previously, and careful attention should be paid during interpretation of CT images in patients with a history of previous percutaneous biopsy. PMID- 10823444 TI - Hepatic periportal tracking associated with severe acute pyelonephritis. AB - The computed tomographic appearance of hepatic perivascular halos has been described in a variety of disorders. We observed three cases with sepsis due to acute pyelonephritis who presented with anasarca and had identical computed tomographic features of periportal edema associated with ascites, pleural effusion, a thickened gallbladder wall, and a dilated inferior vena cava. None of the three patients had an underlying disease process that was previously described as an etiology for an altered hepatic lymphatic dynamics. Acute severe pyelonephritis should be included in the differential diagnosis of extrahepatic diseases that cause hepatic perivascular lucencies. PMID- 10823445 TI - Nodular regenerative hyperplasia of the liver in Budd-Chiari syndrome: CT and MR features. AB - We report the imaging findings of spiral computed tomography (CT), magnetic resonance (MR) imaging, and MR angiography in a patient with nodular regenerative hyperplasia of the liver associated with Budd-Chiari syndrome. Spiral CT showed multiple enhancing nodules during the hepatic arterial and portal venous phases. MR images showed multiple hyperintense nodules on T1-weighted images and hypointense or isointense nodules on T2-weighted images. MR angiography showed thrombotic occlusion of three hepatic veins, suggesting Budd-Chiari syndrome. PMID- 10823446 TI - Resection of liver granulomas under a diagnosis of metastases from breast cancer. AB - Three cases of liver granuloma mistakenly diagnosed as metastases from breast cancer are described. No common cause for granuloma formation in the liver was evident among the patients. Although surgical intervention to obtain a definitive diagnosis may occasionally be necessary, care needs to be exercised in the preoperative diagnosis of liver tumors. The growth rate of the lesions may be an important factor to consider. PMID- 10823447 TI - Hepatocellular carcinoma presenting as portal thrombosis with intrabiliary growth: US and MR findings. AB - We present an unusual case of hepatocellular carcinoma presenting as massive portal vein thrombosis with progression to the intrahepatic bile ducts without demonstrable primary hepatic tumor. Ultrasound, magnetic resonance, and percutaneous transhepatic cholangiography findings are described. The histologic diagnosis was achieved by means of percutaneous forceps biopsy of the endobiliary mass. PMID- 10823448 TI - Intrahepatic biliary calculi: correlation of unusual MR findings with pathologic findings. AB - We report a case of intrahepatic biliary calculi. A localized dilated intrahepatic duct of the left lateral segment of the liver was filled with material that showed marked hyperintensity on T1- and T2-weighted magnetic resonance (MR) images. These MR findings are unusual for intrahepatic stones. Pathologically, the stones were muddy bilirubin calculi, the chemical and physical characteristics of which are surmised to have been the cause of the unusual MR findings. PMID- 10823449 TI - Torsion of the fundus of gallbladder demonstrated on ultrasound and treated with ERCP. AB - A rare case of torsion of the gallbladder fundus, which was diagnosed by ultrasound and endoscopic retrograde cholangiopancreatography (ERCP) and released during ERCP, is presented. The case illustrates the sonographic and ERCP findings of this rare condition and suggests using ERCP as a therapeutic tool. PMID- 10823450 TI - Renal and perirenal space involvement in acute pancreatitis: spiral CT findings. AB - BACKGROUND: This study was conducted to estimate the prevalence and morphologic computed tomographic (CT) features of renal and perirenal space abnormalities in acute pancreatitis in correlation with the severity of pancreatitis. METHODS: One hundred fifty-nine contrast-enhanced CT scans of 100 consecutive patients with acute pancreatitis were retrospectively and independently reviewed by three observers. All CT images were obtained using contrast-enhanced helical CT (collimation width = 5 mm, table increment = 7 mm/s, reconstruction interval = 5 mm, scan delay time = 30-50 s). Additional maximized images (field of view = 260 mm) of the perirenal space were available for review. All CT scans were scored with the CT Severity Score Index: pancreatitis was graded as mild (0-2 points), moderate (3-6 points), and severe (7-10 points). Interobserver agreement for both the severity score and the presence of renal and perirenal involvement was calculated. Correlation between the prevalence of complications and the degree of pancreatitis was estimated. RESULTS: CT scans were graded as mild (n = 59), moderate (n = 82), and severe (n = 18). Abnormalities detected included perirenal stranding (n = 37 patients, 26 bilateral), perirenal fluid collections (n = 10 patients, one bilateral), ureteral encasement (n = 2 patients), renal vein thrombosis (n = 1 patient), and renal parenchymal abnormalities (n = 1 patient). The interobserver agreement range for scoring the degree of pancreatitis and the overall presence of abnormalities was 75.5-79.2% and 59.8-100%, respectively. Except for stranding of the perirenal fat, no statistically significant differences between the presence of abnormalities and the severity of pancreatitis (moderate or severe) was observed with Fisher's exact test. Also, no preferential left-sided localization of complications was observed. CONCLUSIONS: The incidence of renal and perirenal complications from acute pancreatitis is higher than previously estimated (7%). We found no significant correlation between the prevalence of major complications and the severity of pancreatitis. These findings are important because these complications may have an impact on therapeutic strategy and can affect prognosis. PMID- 10823451 TI - Retroperitoneal perforation during endoscopic sphincterotomy: imaging findings. AB - BACKGROUND: Endoscopic retrograde cholangiopancreatography with sphincterotomy has become an important technique in the diagnosis and treatment of biliary and pancreatic diseases. Serious complications, although rare, may occur, and their early recognition and treatment are of the utmost importance. We encountered several such cases. This study reviews the imaging findings in patients with retroperitoneal perforation detected after the procedure. METHODS: Of 796 patients who had endoscopic sphincterotomy at our institution during a 9-year period, retroperitoneal perforation occurred in nine (1.13%). Imaging findings and clinical outcome were assessed. RESULTS: In eight patients, routine abdominal radiographs taken during the procedure disclosed retroperitoneal air, associated with extravasation of contrast material in six patients. This was further confirmed on computed tomography (CT) in three patients. In the ninth patient, the diagnosis was established by an emergent abdominal CT performed a day after the sphincterotomy, because of severe abdominal pain. Two patients died of overwhelming sepsis. CONCLUSIONS: Retroperitoneal perforation during endoscopic sphincterotomy is a rare complication, which occurred in 1.13% of our patients. It can be usually clearly recognized radiographically by an abdominal film and in doubtful cases by CT. We emphasize the importance of recognizing this potentially serious complication with imaging studies. PMID- 10823452 TI - Primary angiosarcoma of the spleen--CT, MR, and sonographic characteristics: report of two cases. AB - Primary angiosarcoma of the spleen is a rare entity, but it is the most common primary splenic malignancy. These tumors demonstrate an aggressive growth pattern and can be single or multiple. The diagnosis should be suspected in a patient who presents with splenomegaly but without evidence of lymphoma, malaria, leukemia, or portal hypertension. The tumor may also present with acute abdominal symptoms secondary to spontaneous splenic rupture. We describe two cases of primary angiosarcoma of the spleen with computed tomographic, magnetic resonance, and sonographic features. PMID- 10823453 TI - Gaucher disease of the spleen: CT and MR findings. AB - We present a 26-year-old male patient with Gaucher disease who presented with epigastric pain and a palpable mass in the left abdomen. Ultrasound, abdominal computed tomography, and magnetic resonance imaging showed massive splenomegaly with multiple splenic nodules up to 7 cm in diameter. Splenic nodules should be included in the differential diagnosis of splenic masses. Follow-up is necessary because of the increased incidence of hematologic malignancies in Gaucher disease. PMID- 10823454 TI - Four-year study of abdominal ultrasound in 900 Central African adults with AIDS referred for diagnostic imaging. AB - BACKGROUND: In the majority of sub-Saharan African countries, the absence of computed tomography facilities makes abdominal ultrasound (US) an alternative diagnostic tool in the clinical investigation of infectious and noninfectious complications of human immunodeficiency virus (HIV)-infected individuals. We studied the abdominal US findings in Central African adult AIDS patients to determine whether the findings were consistent between different population groups and neighboring countries. We performed a longitudinal study of AIDS patients and age- and sex-matched HIV-negative adults referred for abdominal US at two tertiary referral city hospitals: the Gecamines Sendwe Hospital (GSH), Lubumbashi, Congo, and the University Teaching Hospital (UTH), Lusaka, Zambia. METHODS: Between 1992 and 1996, abdominal US findings in 900 adults (300 Congolese adults from GSH and 600 Zambian adults from UTH; age range = 15-55 years) with a diagnosis of AIDS referred for diagnostic imaging from the inpatient medical wards were recorded; 900 abdominal ultrasound findings from age and sex-matched HIV-negative adults were studied for comparative purposes. RESULTS: Abdominal US for diagnostic purposes in AIDS patients is requested by clinicians for a range of primary clinical indications: abdominal pain, fever of unknown origin, hepatosplenomegaly, lymphadenopathy, and abnormal liver function tests. Compared with the HIV-individuals, the AIDS group of patients had a significantly higher proportion of splenomegaly (35% vs. 24%; p < or = 0.001), hepatomegaly (35% vs. 22%; p = 0.001), lymphadenopathy (31% vs. 11%; p < or = 0.001), biliary tract abnormalities (25% vs. 12%; p < or = 0.001), gut wall thickening (15% vs. 5%; p < or = 0.001), and ascites (22% vs. 9%; p < or = 0.001). There were no differences in renal tract and pancreatic abnormalities between the AIDS and HIV-groups. There were significantly fewer gallstones in the AIDS group (23% vs. 75%; p < or = 0.001). These patterns of abdominal US abnormalities were consistent across both hospitals. CONCLUSIONS: Diagnostic imaging by abdominal US is commonly used in the management of a variety of clinical indications in Central Africa. The changes seen on abdominal US in AIDS patients appear uniform across the two countries in Central Africa. These findings may have implications for the radiologist, especially in developing countries, where accurate microbiological or pathologic diagnosis of infectious and noninfectious diseases afflicting the HIV-infected patient is often not possible and US is sometimes relied upon as a "diagnostic" investigation by many physicians. Further studies are required to define patterns of clinical findings, plain films, and pathologic and laboratory correlates with US to develop and refine diagnostic algorithms for clinical use in resource-poor countries. PMID- 10823455 TI - Familial Mediterranean fever: abdominal imaging findings in 139 patients and review of the literature. AB - BACKGROUND: The purpose of this study was to investigate the imaging findings in patients with familial Mediterranean fever (FMF) during and between acute attacks. METHODS: Computerized search of medical records from 1989 to 1998 identified 139 patients with a discharge diagnosis of FMF. Medical records, imaging studies, and pathologic findings were reviewed. RESULTS: Sixty-eight patients had a documented acute attack of FMF, and 71 patients known to have FMF were asymptomatic. Imaging was performed in 68 patients. Radiologic findings included ascites, splenomegaly, hepatomegaly, lymphadenopathy, focal peritonitis, peritoneal cysts, renal changes, and other incidental findings. CONCLUSIONS: Radiologic findings in symptomatic and asymptomatic FMF patients are not uncommon. Imaging in selected cases may facilitate diagnosis and show complications. PMID- 10823456 TI - Postoperative pneumoperitoneum as detected by CT: prevalence, duration, and relevant factors affecting its possible significance. AB - BACKGROUND: To determine the prevalence and duration of postoperative pneumoperitoneum as detected on computed tomography (CT) and to evaluate factors that influence postoperative pneumoperitoneum. METHODS: One hundred three CT examinations of 89 patients performed after abdominal surgery for various indications were prospectively collected and reviewed. The presence and volume of free air were noted and correlated with patients' sex, age, and habitus, with the time interval between surgery and CT, with the type of surgery, and with the presence of drains. RESULTS: Pneumoperitoneum was seen in 44% of examinations performed in the first 3 days after surgery and in 30% between the 4th and 18th postoperative days. The prevalence and volume of free air decreased with the time interval between surgery and CT. It was not detected in any of the 11 examinations performed beyond the 18th postoperative day. The volume of free air in the majority of examinations did not exceed 10 mL and ranged from 0.2 to 10 mL in 19 patients (66% of patients with free air). Larger volumes of free air, ranging from 10 to 20 mL and from 20 to 40 mL, were observed in 5 (17%) and 4 (14%) patients, respectively, and only one patient had more than 40 mL of free air. Free air was found significantly more often in male and in asthenic patients; age had no significant effect. Free air was more prevalent in the presence of drains. The type of surgery did not significantly influence the prevalence of postoperative pneumoperitoneum, although it was noted slightly less often after laparoscopic surgery than after open laparotomy. CONCLUSIONS: Pneumoperitoneum is a common phenomenon after abdominal surgery, decreasing in frequency with time. The air is most often residual and not a sign of disruption of the gastrointestinal tract. Obesity, female sex, and occurrence of free air several weeks after surgery are factors suggestive of a leak, but the significance of a postoperative pneumoperitoneum on CT should be determined mainly by the clinical setting. PMID- 10823457 TI - Mesenteric cyst: sonographic findings. AB - BACKGROUND: Mesenteric cyst (MC) is a relatively rare disease, and its sonographic characteristics have not been sufficiently analyzed. METHODS: We studied the sonographic findings of eight patients with MC, with attention paid to its size, shape, internal echoes, and especially the presence or absence of lateral shadowing and the mode of back echoes. In four cases, the sound velocity and acoustic impedance of cystic fluid were also measured. The mode of blood flow was evaluated by color Doppler sonography. RESULTS: Six cases showed an oval or comma-shaped mass. Internal echoes were present in six cases, and two of them showed a pseudosolid pattern. In these cases, M-mode sonography confirmed the movement of these internal echoes. Only one case showed a posterior echo enhancement, and no case showed lateral shadowing. Sound velocity measured in four cases was 1,515-1,537 m/s, with an acoustic impedance of 1.550-1.576 kg/m2/s. No blood flow signals were obtained from the lesion. CONCLUSION: MC exhibits so many patterns on ultrasound that we should consider the possibility of MC when encountering an avascular oval mesenteric mass. PMID- 10823458 TI - Synchronous mucinous tumors of the ovary and the appendix associated with pseudomyxoma peritonei: CT findings. AB - BACKGROUND: To present the computed tomographic (CT) findings of synchronous mucinous tumors of the ovary and the appendix associated with pseudomyxoma peritonei (PMP). METHODS: Imaging studies, mainly abdominal CT scans, of three women aged 49-75 years were reviewed. Attention was directed to the ovarian masses, peritoneal seeding, and the presence of an appendiceal mucocele. RESULTS: The ovarian tumors and the appendiceal mucocele were clearly demonstrated in two cases, and they were part of the extensive PMP in the third patient. Ascites was found in all cases, with internal septation in one. Associated scalloping of the liver margins and hypodense peritoneal implants, with extensive bowel involvement, were seen in another one. Pathologically, there was one case of right ovarian mucinous cystadenoma and villous adenoma of the appendix, one case of right ovarian and appendiceal mucinous cystadenocarcinoma, and one case of bilateral metastatic ovarian implants of appendiceal mucinous cystadenocarcinoma. PMP was found in all. In the case with benign tumors of the ovary and the appendix, the PMP was classified as a benign mucinous spillage. This patient returned 33 months after surgery with PMP, in which epithelial cells were found. CONCLUSIONS: Radiologists should be familiar with the clinical occurrence of synchronous mucinous tumors of the ovary and the appendix associated with PMP and with the typical CT findings of the latter two entities. Alternatively, when the imaging findings suggest ovarian cystic tumor with PMP, the radiologist should be alerted to the probability of a clinically unsuspected appendiceal mucocele and should search for it. PMID- 10823459 TI - Delayed computed tomographic characterization of renal masses: preliminary experience. AB - BACKGROUND: To evaluate the performance of delayed contrast enhanced computed tomography (DCT) in characterizing renal masses. METHODS: Twenty-four patients with suspected renal masses or indeterminate renal masses on previous imaging studies were prospectively evaluated with preintravenous contrast imaging, conventional contrast-enhanced computed tomography (imaging initiated 2 min after intravenous contrast injection), and DCT (imaging initiated 13 min after injection of intravenous contrast). Only lesions larger than 1.0 cm were evaluated, with scanning parameters kept constant across the three scans. RESULTS: All pathologically confirmed renal cell carcinomas (n = 6) were detected on DCT using a threshold attenuation decrease of 10 Hounsfield units (HU). A significant decrease (p = 0.031) in attenuation occurred in renal cell carcinomas (mean = 29.6 +/- 23.6 HU) compared with the attenuation change (mean decrease = 1.1 +/- 7.1 HU), which occurred in non-neoplastic renal cysts (n = 34). Non neoplastic renal cysts were correctly classified by DCT 32 of 34 times (94%). CONCLUSIONS: In this study, DCT distinguished renal cell carcinomas from non neoplastic cysts in a vast majority of cases and may aid in characterizing incidentally discovered renal lesions on postcontrast CT. PMID- 10823460 TI - Mumps epididymo-orchitis: sonography and color Doppler sonographic findings. AB - Eleven cases of mumps epididymo-orchitis were examined by gray-scale and color Doppler ultrasonography. Nine cases were unilateral and two were bilateral. In the initial examination, the volume and the vascularity of the affected testis and epididymis and the thickness of the scrotal wall was greater than of the normal site, whereas the testicular echogenicity was decreased homogenously, resistivity indexes of intratesticular arteries were decreased, and spontaneous venous flow was seen in all cases. In contrast to nonspecific epididymo-orchitis, no hydrocele was seen, but minimal reactive hydrocele was found in two cases. The diagnosis was confirmed by specific immunoglobulin-G examination. Patients were given interferon and were controlled by ultrasonography and Doppler ultrasonography at the third and seventh days of treatment and 3 months after treatment. Sonographic findings began to improve by the third day and fully disappeared in seventh day. No testicular atrophy was seen in the last control. To our knowledge, this is the first report on sonographic and color Doppler sonographic findings of mumps epididymo-orchitis. PMID- 10823461 TI - Damage to liposomal lipids: protection by antioxidants and cholesterol-mediated dehydration. AB - Liposomes composed of egg phosphatidylcholine (EPC) (13.4%, of the acyl chains being polyunsaturated fatty acids (PUFA)) and EPC/cholesterol (10:1 mol/mol) were studied for factors that affect liposomal lipid oxidative damage and hydrolysis upon long-term (16 months) storage. Factors studied include: (1) levels of lipid/water interface hydration, related to the presence of cholesterol in the lipid bilayer; (2) the membrane-associated antioxidant vitamin E; (3) the water soluble antioxidant Tempol; and (4) exposure to light. Liposomal dispersions were stored at room temperature, either exposed to or protected from daylight, for a period of 16 months. Chemical and physical changes were monitored at several time points to assess oxidative and hydrolytic degradation of liposomal lipids. The conclusions of the study are: (1) PUFA are the most sensitive component of the liposome bilayer to oxidative degradation damage during long-term storage; (2) EPC liposomes are more sensitive to degradation during storage than EPC cholesterol liposomes, the presence of cholesterol in the lipid bilayer having a protective effect, probably due to its effect in decreasing the lipid-bilayer hydration; (3) oxidative degradation is the major process during long-term storage, having an earlier onset than the hydrolytic degradation: and (4) Tempol provided significantly better protection than vitamin E to EPC liposomal PUFA against oxidative damage during long-term storage. The relevance of cholesterol's presence, as a 'drying agent', in membranes containing PUFA to resistance of biological membranes to oxidative damage is discussed. PMID- 10823462 TI - Giant phospholipid vesicles: comparison among the whole lipid sample characteristics using different preparation methods: a two photon fluorescence microscopy study. AB - Several methods for the preparation of giant unilamellar vesicles (GUVs) using synthetic phosphatidylcholine phospholipids were evaluated. We compared the physical characteristics--in terms of lamellarity and morphology--of the whole lipid sample for each different lipid preparation using the sectioning capability of the two-photon excitation fluorescence microscope. From the evaluation of the entire lipid sample we determined that vesicle size, internal shape and shell thickness distributions depend on the vesicle's preparation method. Our results show that the preparation of giant unilamellar vesicles by the application of external electric fields offers several advantages among the other methods tested here. Using this method a high yield (approximately 95%) of giant unilamellar vesicles with a narrow size distribution was obtained. Independently of the preparation method, some lipid structures, which are held together by lipid tethers, were identified and resolved. These particular lipid structures show shell thickness and size heterogeneity. Labeling the lipid samples with 6-lauroyl 2-(N,N-dimethylamino)naphtalene (LAURDAN) and using the LAURDAN generalized polarization function we show that the lipid packing in these tethers or tubes is similar to those found in the phospholipid vesicles. The fact that both vesicles and tethers are found in the lipid preparations indicates similar stability between these structures. PMID- 10823463 TI - Hydration of DMPC and DPPC at 4 degrees C produces a novel subgel phase with convex-concave bilayer curvatures. AB - Hydration of dimyristoyl- and dipalmitoylphosphatidylcholines at 4 degrees C results in the formation of a characteristic subgel phase designated Pcc. Examination of the phase by freeze-fracture electron microscopy shows convex concave deformations of the planar bilayer which are of two types. A smaller type with a radius of curvature of about 20 nm predominates in DMPC, and a larger type with about 70 nm radii of curvatures dominates in DPPC. The Pcc phase can also be formed in samples hydrated at temperatures above the main phase transition if the dispersion is frozen slowly and subsequently incubated at 4 degrees C for several days. The subgel Pcc phase was distinguished from the subgel Lc phase by the temperature of transition, packing of the acyl chains on the basis of wide-angle X-ray diffraction, and 2H-NMR spectra characteristic of a 'solid-ordered' phase. Vibrational spectra of the carbonyl and phosphate regions are consistent with a partially reduced hydration state. The origin of the convex-concave bilayer deformation is believed to result from constraints imposed by limiting hydration of the headgroup and a frustration arising from the spontaneous curvature of both monolayers. PMID- 10823464 TI - Cyclodextrin-catalyzed extraction of fluorescent sterols from monolayer membranes and small unilamellar vesicles. AB - This study examined the kinetics of sterol desorption from monolayer and small unilamellar vesicle membranes to 2-hydroxypropyl-beta-cyclodextrin. The sterols used include cholesterol, dehydroergosterol (ergosta-5,7,9,(11),22-tetraen-3beta ol) and cholestatrienol (cholesta-5,7,9,(11)-trien-3beta-ol). Desorption rates of dehydroergosterol and cholestatrienol from pure sterol monolayers were faster (3.3-4.6-fold) than the rate measured for cholesterol. In mixed monolayers (sterol: 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine 30:70 mol%), both dehydroergosterol and cholestatrienol desorbed faster than cholesterol. clearly indicating a difference in interfacial behavior of these sterols. In vesicle membranes desorption of dehydroergosterol was slower than desorption of cholestatrienol, and both rates were markedly affected by the phospholipid composition. Desorption of sterols was slower from sphingomyelin as compared to phosphatidylcholine vesicles. Desorption of fluorescent sterols was also faster from vesicles prepared by ethanol-injection as compared to extruded vesicles. The results of this study suggest that dehydroergosterol and cholestatrienol differ from cholesterol in their membrane behavior, therefore care should be exercised when experimental data derived with these probes are interpreted. PMID- 10823465 TI - Antioxidant activities of estrogens against aqueous and lipophilic radicals; differences between phenol and catechol estrogens. AB - Natural estrogens have much greater radical-scavenging antioxidant activity than has previously been demonstrated, with activities up to 2.5 times those of vitamin C and vitamin E. The biological significance of this finding remains to be elucidated. In this work the antioxidant activity of a range of estrogens (phenolic, catecholic and stilbene-derived) has been studied. The activity of these substances as hydrogen-donating scavengers of free radicals in an aqueous solution has been determined by monitoring their relative abilities to quench the chromogenic radical cation 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS*+). The results show that the order of reactivity in scavenging this radical in the aqueous phase is dependent on the precise estrogenic structure, with phenolic estrogens being more potent antioxidants than catecholestrogens or diethylstilbestrol. The ability of the same estrogens to scavenge lipid phase radicals has also been assessed, determined by the ex vivo enhancement of the resistance of low-density lipoprotein (LDL) to oxidation; the order of efficacy is different from that in the aqueous phase, with the phenolic estrogens estriol, estrone and 17beta-estradiol being less potent than 2-hydroxyestradiol, 4 hydroxyestradiol, or diethylstilbestrol. In this lipid-based system, phenolic estrogens were found to be unable to regenerate alpha-tocopherol from LDL subjected to oxidative stress, while at the same time 2- and 4-hydroxyestradiol significantly delayed alpha-tocopherol loss. These results indicate that the various estrogens are good scavengers of free radicals generated in both the aqueous and the lipophilic phases. The antioxidant activity of an estrogen depends not only on the hydrophilic or lipophilic nature of the scavenged radical, but also on the phenol and catechol structures of the estrogen compound. PMID- 10823466 TI - A simple approach to DOTAP and its analogs bearing different fatty acids. AB - A simple synthesis of N-[1-(2,3-dioleoyloxy)propyl]-N,N,N-trimethylammonium methyl sulfate (DOTAP) and its analogs differing in the fatty acids is presented. The synthesis is designed as quasi-one-pot reaction and the precipitating products are purified by simple recrystallization. PMID- 10823467 TI - Molecular modeling of archaebacterial bipolar tetraether lipid membranes. AB - Membranes composed of glycerol dialkylnonitol tetraether (GDNT) lipids from the thermoacidophilic archaebacterium Sulfolobus acidocaldarius have been studied by molecular modeling. GDNT membranes containing eight cyclopentane rings in the molecule are packed much tighter than those without rings. When containing eight cyclopentane rings, the beta-D-galactosyl-D-glucose head-group of GDNT runs almost parallel to the membrane surface. However, when containing no rings, the head-group is oriented perpendicular to the membrane surface. Using molecular dynamics calculations, we have also conducted comparative studies of membrane packing between GDNT and various non-archaebacterial membranes. Compared to gel state dipalmitoylphosphatidylcholine (DPPC) and gel state distearoylphosphatidylcholine (DSPC) bilayers, the GDNT membrane with eight cyclopentane rings has a more negative interaction energy, thus a tighter membrane packing, while the GDNT without rings is less tightly packed than gel state DSPC. Based on the calculated interaction energies, the GDNT membranes (with and without rings) are much more tightly packed than DPhPC (an ester-linked diphytanyl PC) and DPhyPC (an ether-linked diphytanyl PC) bilayers. This suggests that the branched methyl group in the phytanyl chain is not the major contributor of the tight packing of GDNT membranes. The biological implication of this study is that the cyclopentane ring could increase GDNT membrane thermal stability. This explains why the number of cyclopentane rings in archaebacterial lipid increases with increasing growth temperature. Perhaps, through the ring temperature compensation mechanism the plasma membrane of thermoacidophilic archaebacteria is able to maintain a tight and rigid structure, consequently, a constant proton gradient between the extracellular (pH 2.5) and intracellular compartment (pH 6.5), over a wide range of growth temperatures. PMID- 10823468 TI - Phase transition in dioctadecyldimethylammonium bromide and chloride vesicles prepared by different methods. AB - The gel to liquid crystalline phase transition of the double-chained cationic dioctadecyldimethylammonium chloride and bromide (DODAX, X = Cl- or Br-) in aqueous vesicle dispersions prepared by non-sonication. sonication and extrusion has been investigated using high-sensitivity differential scanning calorimetry (DSC). The transition temperature (Tm) is a function of the preparation method, amphiphile concentration, vesicle curvature and nature of the counterion. DSC thermograms for DODAB and DODAC non-sonicated vesicle dispersions exhibit a single endothermic peak at Tm roughly independent of concentration up to 10 mM. Extrusion broadens the transition peak and shifts Tm downwards. Sonication, however, broadens slightly the transition peak and tends to shift Tm upwards suggesting that extrusion and sonication form vesicles with different characteristics. DODAC always exhibits higher Tm than DODAB irrespective of the preparation method. Tm changes as follows: Tm (sonicated) > or = Tm (non sonicated) > Tm (extruded). Hysteresis of about 7 degrees C was observed for DODAB vesicle dispersions. PMID- 10823469 TI - Antioxidants eliminate stereomutation and thioether formation during lipase catalyzed thioesterification and transthioesterification for the preparation of uniform cis- and trans-unsaturated thioesters. AB - The lipase-catalyzed preparation of acyl thioesters from unsaturated fatty acids and alkanethiols is accompanied by the formation of geometrical isomers via stereomutation and of thioether derivatives via addition at the olefinic bond, both induced by thiyl radicals. Therefore, a method was developed in order to inhibit radical generation by the addition of antioxidants and thus prevent the formation of geometrical isomers and thioether derivatives during the lipase catalyzed preparation of unsaturated acyl thioesters. In the presence of antioxidants such as 2,6-di-t-butyl-4-methylphenol (BHT) and octyl gallate thioesterification of oleic and elaidic acids with 1-tetradecanethiol as well as transthioesterification of methyl linoleate with 1-tetradecanethiol led to the corresponding geometrically uniform thioesters without radical-induced side reactions. In the absence of antioxidants rapid stereomutation of unsaturated acyl moieties as well as formation of high proportions of thiyl radical-induced addition products such as isomeric 9(10)-S-tetradecyl stearic acids and 9(10)-S tetradecyl stearic acid tetradecyl thioesters were observed. PMID- 10823470 TI - Double layer potential and degree of dissociation in charged lipid monolayers. AB - One of the contributions to the surface potential in charged phospholipid monolayers at air-water interfaces is the double layer potential. In this note several misconceptions found in the literature concerning the relationship between the double layer potential and the degree of dissociation of the lipid polar headgroups are critically analyzed. The deviations of the double layer potential measurements from the Gouy-Chapman theory observed by several authors are explained by taking into account the dependence of the degree of dissociation with concentration, area per lipid molecule and pH. PMID- 10823471 TI - Glottal area waveform analysis of benign vocal fold lesions before and after surgery. AB - Glottal area waveform (GAW) is the plot of relative glottal area versus time through 1 representative glottal cycle. It is derived from the quantitative analysis of the videostroboscopic image. A GAW analysis was performed on 24 patients before and after microlaryngeal phonosurgery. Patients with vocal fold polyps, polypoid degeneration, cysts, sulcus vocalis, and Reinke's edema were included. From each GAW, 5 parameters were determined and compared: maximum normalized glottal area, maximum opening rate, maximum closing rate, percent open time at 50% glottal opening, and glottal gap size. Statistically significant differences in postoperative states included an increased maximum glottal area, an increased maximum opening rate, and an increased maximum closing rate. Glottal opening and closing rate are objective measures of vocal fold pliability that have clinical relevance. The GAW may be used to quantitate vocal fold vibratory capability. PMID- 10823472 TI - Laryngeal adductory pressure as a measure of post-reinnervation synkinesis. AB - Laryngeal adductory pressure (LAP) is the pressure induced as the vocal folds squeeze on a balloon while the recurrent laryngeal nerve (RLN) is stimulated. The LAP has been shown to vary with the frequency of stimulation, with a characteristic slope. The RLN was divided and reanastomosed 4 different ways in 12 canine hemilaryngeal preparations; the 4 subgroups represented a range of expected post-reinnervation synkinesis recovery patterns. The LAP frequency response curve was measured before surgery and at monthly intervals for 6 months after surgery. In the "best-case" group (RLN adductor and abductor trunks each divided and reanastomosed), the slope was found to return to normal. The 2 whole RLN division-reanastomosis groups (precise realignment or 180 degrees rotation) both gave results similar to those of the "worst-case" group (RLN adductor and abductor trunks divided and transposed); these 3 subgroups were all significantly different from baseline. The slope of the LAP frequency-response curve may be a useful means of indirectly quantifying laryngeal synkinesis. PMID- 10823473 TI - Pyriform sinus fistula: management with chemocauterization of the internal opening. AB - A branchial remnant originating in the pyriform sinus causes a recurrent fistula or abscess in the neck. In spite of excision, recurrence may result from inadequate removal of the fistula tract. We attempted chemocauterization of the internal opening of the fistula tract with trichloroacetic acid (TCA) on direct endoscopy. This is a 6-year review of 18 patients with pyriform sinus fistula. Medical history, barium esophagography, computed tomography scans, operative findings, and treatment results were analyzed. By direct endoscopy, all patients were found to have a fistula opening in the pyriform sinus, exclusively on the left side. In only 9 patients, the fistula tract was identified by barium esophagography before operation. Computed tomography revealed a suspicious fistula tract originating from the pyriform sinus in 8 of 10 patients. Sixteen patients were initially managed by TCA chemocauterization. There were no serious intraoperative or postoperative complications. Four patients had recurrent masses, which were managed by simple excision in 2 patients and repeated TCA cauterization in the other 2 patients with unobliterated internal openings. We recommend barium swallow study and direct endoscopy for all patients presenting with a recurrent lateral neck abscess, especially on the left side. Our results suggest that initial chemocauterization of the internal opening can be a reasonable alternative procedure for the management of pyriform sinus fistula. PMID- 10823474 TI - Experimental autoimmune inner ear disease: an electrocochleographic and histophysiologic study. AB - Systemic immunization with swine inner ear antigens in complete Freund's adjuvant induces functional disturbances in the cochlea. Morphometric data indicate that an endolymphatic hydrops develops within 2 weeks. It diminishes 6 weeks after immunization. A progressive decrease in the compound action potential amplitude is observed from 2 to 6 weeks after immunization. Enhancement of the amplitude of the summating potential is present without a clear overall correlation to the presence of endolymphatic hydrops. The amplitude of the cochlear microphonics shows no significant changes after immunization. Western blot analysis of the sera performed 2 and 6 weeks after immunization shows enhanced reactivity at 68, 50, 45, and 27 kd molecular weights, as compared to controls. The same spectrum of cross-reacting antibodies is believed to be instrumental in immune-mediated sensorineural hearing loss in patients. Apparently, cross-reacting antibodies and released mediators disturb cochlear homeostasis, resulting in the observed changes in the electrophysiological responses. However, these changes are not clearly related to structural changes at the light and electron microscopic levels. PMID- 10823475 TI - Cellular distribution of mucosa-associated lymphoid tissue with otitis media in children. AB - This study examined mucosa-associated lymphoid tissue (MALT) in the eustachian tube (ET), middle ear (ME), and mastoid antrum (MA) in 163 celloidin-embedded temporal bones from children with or without otitis media. Otitis media was defined by the presence of histopathologically identified inflammatory cell infiltration in the mucosa or cavity of the ME. We found MALT in the ET in 30 cases (46.2%), in the ME in 19 cases (29.2%), and in the MA in 4 cases (6.2%) out of 65 cases of otitis media, and in the ET in 7 (7.1%), in the ME in 0, and in the MA in 0 out of 98 specimens without otitis media. No MALT appeared in any children under the age of 1 month. Immunohistochemical methods were used to investigate MALT in 12 horizontally cut temporal bones with OM. The follicular area contained OPD4-positive (helper-inducer T) cells and a few CD8-positive (cytotoxic and suppressor T) cells, whereas the parafollicular area contained OPD4-positive and CD8-positive T cells. CD57-positive (natural killer) cells were confined to the germinal center. CD30-positive (activated T and B) cells were observed throughout the follicles. A few CD15-positive (granulocyte, monocyte) cells were found in the follicles. Histopathologic and immunohistochemical findings were indistinguishable for MALT in the ET, ME, and MA. Our results suggest that MALT may be a mechanism for producing a rapid and massive local immune reaction to repeated bacterial infections via the ET. PMID- 10823476 TI - Assessment of stapes mobility by use of a newly developed piezoelectric ceramic device. A preliminary experiment in dogs. AB - A piezoelectric device was developed for assessment of stapes mobility during middle ear surgery. The device comprises a pair of ceramic bimorph elements: one for activation of the stapes and the other to pick up the vibration as an electric output, which varies in accordance with the stapes mobility, ie, the inverse of the cochlear input impedance (Zsc). The device is compact and easily manipulated even in the narrow surgical field of the ear. However, the measuring range is restricted to between 1 and 10 kHz. Measurement of Zsc was conducted with this device in 5 ears of 5 dogs. The mean magnitude of Zsc increased with frequency in the range from 1 to 10 kHz: 0.95 megohm at 1 kHz and 8.8 megohms at 10 kHz. After fixation of the stapes with dental cement, the magnitude increased to more than 10 megohms, except at 1 kHz. The results suggest that the device is useful in detecting decreases in stapes mobility in patients with chronic otitis media. PMID- 10823477 TI - Quantification of the three-dimensional displacement of normal facial movement. AB - This study was undertaken to quantify 3-dimensional (3-D) facial movement in normal subjects, and to identify the individual axes in which this movement occurred. Displacement data on 42 subjects were collected and analyzed with the Expert Vision Motion Analysis System. The 3-D displacement was calculated by vectorially subtracting maximum marker movement from previously identified reference marker points. The 3-D range of normal facial movement was quantified, with the greatest displacement occurring during maximum smile. When the individual axes were examined, we found that most movement occurred in the vertical axes for the majority of expressions, followed by the anterior-posterior axis. These results may create an objective baseline from which disorders of the facial nerve, and hence, medical, surgical, and physiotherapy treatment interventions, can be analyzed in the future. PMID- 10823478 TI - Effects of clarithromycin on rheological properties of nasal mucus in patients with chronic sinusitis. AB - Macrolide antibiotics have a variety of actions along with antimicrobial action. To determine the effects of oral administration of clarithromycin (CAM) on rheological properties, we measured the spinability, dynamic viscoelasticity, and solid composition of human nasal mucus from 18 patients with chronic sinusitis before and after administration of CAM for 4 weeks. After administration of CAM, the spinability and percent solid composition of nasal mucus increased from 26.5 +/- 12.2 mm to 40.2 +/- 18.7 mm and 7.86% +/- 3.47% to 13.90% +/- 3.67% (p < .05), respectively, whereas the ratio of the viscosity to the elasticity (eta'/G') of nasal mucus after the administration of CAM decreased in all of the mucus samples. These results suggest that treatment with CAM may modulate the rheological properties of nasal mucus in patients with chronic sinusitis. PMID- 10823479 TI - Human uvula: characterization of resident leukocytes and local cytokine production. AB - Upper airway infections often lead to macroscopic changes in the architecture of the uvula. Using immunomorphometric analysis, we investigated the frequency and distribution of immune cells and of cytokine-producing cells in uvular samples. Tissue macrophages, alphabeta T cells, gammadelta T cells, and B cells were, in declining order, the main cell populations. Gammadelta T cells and B cells exhibited reciprocal localization, with almost all gammadelta T cells residing in the vicinity of the epithelium, and all B cells in the glandular area. The presence of cells expressing the suppressor phenotype CD8+CD28- alphabeta T cells is suggested. Fifteen to twenty-five percent of the immune cells expressed the down-regulatory cytokine tumor growth factor beta. Most macrophages were located subepithelially, in the vicinity of the basal lamina. The composition and cytokine profile of leukocytes in the tissue suggest that the uvula may be a site, additional to the jejunal mucosa, for induction of mucosal tolerance to inhaled and ingested antigens. Concomitantly, the uvula appears to be protected from invasion of microbial pathogens by a subepithelial barrier of macrophages and gammadelta T cells. PMID- 10823480 TI - Congenital defects of the vomer. AB - Six cases of congenital defect of the vomer, a rare nasal anomaly, are reported. All 6 patients visited Kobe University Hospital with other complaints, and the anomaly was incidentally detected. In all cases, the nasal septum showed a defect at the posteroinferior portion that appeared to coincide with the location of the vomer. None of the patients had a past history of nasal trauma, nasal surgery, drug abuse, or infectious disease. This anomaly may be attributable to an embryological disorder based on an immature ossification center of the vomer. PMID- 10823481 TI - Estimated subglottic pressure in normal and dysphonic subjects. AB - The objective of this report was to study the clinical interest of estimated subglottic pressure (ESGP) with measurements of intraoral pressure according to the "airway interrupted method." Twenty healthy female subjects and 27 dysphonic female patients were included and asked to produce sounds under different conditions of pitch and intensity. The ESGP increased with intensity and slightly with pitch in both patients and controls. A comparison between patients and controls showed significantly higher values of ESGP in patients under all conditions of intensity and pitch. For normal intensity and usual pitch, ESGP has been found to be 6.1 hectopascals (hPa) in control subjects and 8.25 hPa in patients (p = .002). Discriminant analysis of all the measured data showed that data recorded for low intensity (lowest possible intensity without whispering) and high pitch (9 semitones above the usual pitch) were the most discriminant. The authors concluded that ESGP allows good discrimination between dysphonic patients and control subjects and might be included in the basic clinical set of objective parameters. PMID- 10823482 TI - Three-dimensional anatomic characterization of the canine laryngeal abductor and adductor musculature. AB - The biomechanics of vocal fold abduction and adduction during phonation, respiration, and airway protection are not completely understood. Specifically, the rotational and translational forces on the arytenoid cartilages that result from intrinsic laryngeal muscle contraction have not been fully described. Anatomic data on the lines of action and moment arms for the intrinsic laryngeal muscles are also lacking. This study was conducted to quantify the 3-dimensional orientations and the relative cross-sectional areas of the intrinsic abductor and adductor musculature of the canine larynx. Eight canine larynges were used to evaluate the 3 muscles primarily responsible for vocal fold abduction and adduction: the posterior cricoarytenoid, the lateral cricoarytenoid, and the interarytenoid muscles. Each muscle was exposed and divided into discrete fiber bundles whose coordinate positions were digitized in 3-dimensional space. The mass, length, relative cross-sectional area, and angle of orientation for each muscle bundle were obtained to allow for the calculations of average lines of action and moment arms for each muscle. This mapping of the canine laryngeal abductor and adductor musculature provides important anatomic data for use in laryngeal biomechanical modeling. These data may also be useful in surgical procedures such as arytenoid adduction. PMID- 10823483 TI - Prevention of pharyngoesophageal spasm after laryngectomy with a half-muscle closure technique. AB - Because pharyngoesophageal spasm can limit successful voice restoration after total laryngectomy, 24 patients underwent a modified pharyngeal closure in an effort to avoid this problem. All patients underwent total laryngectomy with appropriate neck dissections and pharyngeal closure with a half-muscle closure technique in which only one side of the remnant constrictor muscles was used to reinforce the primary closure. Twenty-three patients underwent voice restoration. Twenty-two (96%) had a functional voice, and 1 patient (4%) had pharyngoesophageal spasm that required a secondary myotomy. One fistula (4%) occurred and resolved with conservative measures. Quantitative voice analysis demonstrated no significant differences between half-closure patients and control patients for fundamental frequency (96 Hz versus 101 Hz) or intensity (57 dB versus 64 dB). Extensive qualitative analysis by trained and naive listeners revealed no differences. This preliminary report indicates the half-muscle closure modification of the pharyngeal closure at primary laryngectomy may provide a simple and effective means of avoiding pharyngoesophageal spasm and maintaining an effective voice without increased complications. PMID- 10823484 TI - Pneumomediastinum due to Venturi jet ventilation used during microlaryngeal surgery in a previously neck-irradiated patient. AB - Serious complications secondary to Venturi jet ventilation used during microlaryngoscopy are rare, but when they occur, they may pose a life-threatening emergency. We report the case of a 45-year-old woman, previously treated with 70 Gy of irradiation for a T1 laryngeal carcinoma, who developed pneumomediastinum and subcutaneous emphysema after the use of Venturi jet ventilation. Keeping in mind the histologic changes to the irradiated structures, we suggest more caution when using Venturi jet ventilation in patients who have recently undergone neck irradiation therapy. PMID- 10823485 TI - Mucocele of the infratemporal fossa as an unusual complication of midfacial fracture. PMID- 10823486 TI - Medical management of acute bacterial sinusitis. Recommendations of a clinical advisory committee on pediatric and adult sinusitis. AB - Acute sinusitis is commonly encountered in clinical practice and treated in the primary care setting. The clinician should recognize the subtle clinical presentation of acute bacterial sinusitis and initiate appropriate, aggressive treatment. Other upper respiratory tract disorders can confound the accurate diagnosis and appropriate treatment of sinusitis. Variable patterns of microbial resistance and antibiotic susceptibility and the dissociation between in vitro findings and clinical efficacy are a treatment challenge. This report is a comprehensive review of the pathophysiology and diagnosis of acute sinusitis, infectious agents, treatment methods, antibiotic resistance patterns, and costs associated with the management of sinusitis. Treatment algorithms are presented for adult and pediatric sinusitis. PMID- 10823488 TI - The chemical synthesis of peptides: a select bibliography. PMID- 10823487 TI - Developments in bread-making processes. AB - An introduction to the evolutionary history of wheat is followed by a description of the origin and distribution of the modern hexaploid wheat varieties in Europe. Next, the influence of cultural (the way of bread making), social and economic (i.e. the great influence of the United States and Canada) factors on wheat growing in Europe are considered (Section 1). The anatomy and chemical composition of wheat grains and the fundamental differences between hard and soft wheat are described in Section 2. The process of flour making is treated in Section 3. Section 4 contains a study of gluten proteins and wheat starches in relation to the making of bread (Section 5) and of other wheat products (Section 6). Section 7 deals with the genetic bread-wheat properties: grain hardness, milling quality, production of (unwanted) sticky doughs, alpha-amylase levels, and contains an extensive study of genetics and the role of gliadins and glutenins. A concluding section contains the description and evaluation of assay methods to identify the characteristics of bread wheats for the use of millers, arbiters, and especially for growers. PMID- 10823489 TI - Hydrogen peroxide for disulfide bridge formation in methionine-containing peptides. AB - Two methionine-containing peptides, endothelin 1 and the 1-16 fragment of the receptor of the plasminogen activator 1 for human urokinase, were synthesized and cyclized by hydrogen peroxide. Endothelin 1 was obtained by using regioselective and random schemes of disulfide bond formation. The conditions of cyclization that provided the target products in high purity were found. The general potential of disulfide bond formation by means of hydrogen peroxide was demonstrated for methionine-containing peptides. The method resulted in target products containing insignificant quantities of the corresponding Met-sulfoxide derivatives. PMID- 10823490 TI - Immunogenicity comparison of a multi-antigenic peptide bearing V3 sequences of the human immunodeficiency virus type 1 with TAB9 protein in mice. AB - The multiple antigenic peptide system (MAP) has been proposed as a novel and valuable approach for eliciting antibodies for peptides and developing synthetic vaccines. Multi-epitope polypeptides (MEP) have also been developed as an alternative to the recombinant approach for vaccines. The V3 loop from the HIV type 1 (HIV-1) external glycoprotein (gp120) contains the principal neutralization domain (PND). Antibodies against this region neutralize HIV-1 in vitro and in vivo. In this work, a novel presentation of di-epitope MAP was synthesized. A monomeric MAP carrying two identical JY1 V3 sequences as B-cell epitopes and the 830-843 region of tetanus toxoid as a T-helper cell epitope was synthesized. This basic structure was covalently linked to produce a four-JY1 branched homodimer (JY1-MAP4). Additionally, six different monomeric MAPs, bearing four copies of V3 from isolates LR150, JY1, RF, MN, BRVA and IIIB, were synthesized. These monomers were conveniently linked among themselves to produce homodimeric and heterodimeric MAPs of eight V3 branches (V3-MAP8). JY1-MAP8 elicited higher antibody titers in Balb/c mice than JY1-MAP4. The immunogenicity of two different, hexavalent V3-MAP8 mixtures and the MEP TAB9, which tandems the same six V3 sequences in a single molecule, were compared. The antibody response against the mixtures of the heterodimeric MAP showed a wider recognition pattern of the V3 region, while the homodimeric cocktail showed an intermediate pattern. Antibodies elicited by TAB9 recognized only the JY1, LR150 peptides. These results emphasize the influence of V3 epitope presentation upon the characteristics of the antibody response generated. PMID- 10823491 TI - Peptide thioester preparation by Fmoc solid phase peptide synthesis for use in native chemical ligation. AB - Established methodology for the preparation of peptide thioesters requires the use of t-butoxycarbonyl chemistry owing to the lability of thioester linkers to the nucleophilic reagents used in Fmoc solid phase peptide synthesis. Both the greater ease of use and the broad applicability of the method has led to the development of an Fmoc-based methodology for direct peptide thioester synthesis. It was found that successful preparation of a peptide thioester could be achieved when the non-nucleophilic base, 1,8-diazabicyclo[5.4.0]undec-7-ene, together with 1-hydroxybenzotriazole in dimethylformamide, were used as the N(alpha)-Fmoc deprotection reagent. Native chemical ligation of the resulting thioester product to an N-terminal cysteine-containing peptide was successfully performed in aqueous solution to produce a fragment peptide of human alpha-synuclein. The formation of aspartimide (cyclic imide) in a base-sensitive hexapeptide fragment of scorpion toxin II was found to be significant under the deprotection conditions used. However, this could be controlled by the judicious protection of sensitive residues using the 2-hydroxy-4-methoxybenzyl group. PMID- 10823492 TI - Novel strategy for the synthesis of template-assembled analogues of rat relaxin. AB - The 'template-assembled synthetic protein' (TASP) concept provides a simple and elegant approach for the preparation of analogues that retain key structural elements. We have synthesized TASP molecules containing the putative active site of relaxin, a peptide that has similar structural features to insulin but a markedly different biological role. Two types of chemoselective thiol ligation strategies (thioether and thiazolidine) were used and compared. The synthetic pendant peptides contain an essential region for bioactivity that is located in the alpha-helical region of the relaxin B-chain. Depending on whether the thioether or the thiazolidine chemistry was used to attach the peptides to the template, the reacting amino acid was placed either at the C-terminus or N terminus, respectively, thus allowing the choice of orientation relative to the carrier molecule. The template molecule consists of a decapeptide with two proline-glycine turns and four evenly spaced lysine residues that were functionalized with the appropriate chemical moiety. This allowed reaction with the appropriately derivatized peptides in solution. To improve the template ligation step using the thioether approach, a pendant peptide C-terminal cysteamine residue was used to reduce potential steric hindrance during conjugation. The design of the peptides as well as the synthetic strategy resulted in the acquisition of mimetics showing weak non-competitive and weak competitive antagonist properties. PMID- 10823493 TI - Solid phase synthesis of C-terminal peptide amides: development of a new aminoethyl-polystyrene linker on the Multipin solid support. AB - A new aminoethyl-polystyrene linker, stable at low concentrations of TFA, has been developed for the solid phase synthesis of peptide amides. The described linker is stable under conditions which remove Bu(t) protecting groups (30-50% TFA in DCM) and the desired product can be finally cleaved off the solid support in 95% TFA (5% H2O). Model peptide amides and other N-alkylated peptide amides have been successfully synthesized in good yield and purity. PMID- 10823494 TI - Three-dimensional spatial compounding of ultrasound scans with weighting by incidence angle. AB - A three-dimensional (3D) ultrasound imaging system has been used to study spatial compounding of images acquired with different scanhead positions and orientations. A compounding algorithm has been developed that assigns regional weights depending on the local incidence angle of the ultrasound beam. Compound scans were performed of bones in vitro and the shoulder rotator cuff in volunteer subjects. Border measurements (peak value and width) were compiled as a function of ultrasound beam incidence angle and compared for single views and for maximum, mean and weighted mean compounding techniques. The weighted mean produces less variability than that of the maximum and mean for both intensity and border width. The weighted method also demonstrates less blurring of borders than the maximum and mean methods. Surfaces derived from the weighted reconstructions exhibited fewer gaps and fewer spurious connections between surfaces, which could be of particular importance for automated image analysis. PMID- 10823495 TI - Multiparametric attenuation and backscatter images for characterization of carotid plaque. AB - The goal of this study was to develop methods for quantitative ultrasound imagery suitable for noninvasive assessment of carotid plaque composition prior to the selection of the technique for revascularization. Using two broadband transducers (5-12 MHz and 12-28 MHz), backscattered radio frequency signals were acquired from entire lengths of 15 carotid endarterectomy specimens. Spectral analysis methods with correction for system response and beam diffraction were applied to radio frequency signals from local volumes of plaque having a 2 mm slice thickness, 1 mm width and axial depth of 480 microm and 240 microm at 10 MHz and 20 MHz, respectively. From these spectra, local values of four ultrasound parameters (integrated backscatter, frequency dependence of backscatter, integrated attenuation and slope of attenuation) were estimated and used to construct quantitative images. To combine information from these different parameter images, a two-step approach was followed. First, in 59 independent quantitative images of highly stenotic plaque, the average parameter values in a central five-by-three pixel region were correlated with plaque composition as assessed by histology to investigate the relationship between parameter values, frequency bandwidth and plaque composition. Discriminant analysis of parameter values vs. plaque composition was made to find a set of predictive equations to classify sets of measurements. Correct classification was obtained for 100% of calcified, 75% of intraplaque hemorrhage and 71% of lipidic plaques of the input data set. Second, each set of pixels from different parameter images was classified using the predictive equations, and a single, local tissue composition image was constructed. Examples of tissue composition images are presented in comparison with corresponding histologic sections. Both agreement and disagreement between image pairs are discussed. PMID- 10823496 TI - A finite element model of remote palpation of breast lesions using radiation force: factors affecting tissue displacement. AB - The early detection of breast cancer reduces patient mortality. The most common method of breast cancer detection is palpation. However, lesions that lie deep within the breast are difficult to palpate when they are small. Thus, a method of remote palpation, which may allow the detection of small lesions lying deep within the breast, is currently under investigation. In this method, acoustic radiation force is used to apply localized forces within tissue (to tissue volumes on the order of 2 mm3) and the resulting tissue displacements are mapped using ultrasonic correlation based methods. A volume of tissue that is stiffer than the surrounding medium (i.e., a lesion) distributes the force throughout the tissue beneath it, resulting in larger regions of displacement, and smaller maximum displacements. The resulting displacement maps may be used to image tissue stiffness. A finite-element-model (FEM) of acoustic remote palpation is presented in this paper. Using this model, a parametric analysis of the affect of varying tissue and acoustic beam characteristics on radiation force induced tissue displacements is performed. The results are used to evaluate the potential of acoustic remote palpation to provide useful diagnostic information in a clinical setting. The potential for using a single diagnostic transducer to both generate radiation force and track the resulting displacements is investigated. PMID- 10823497 TI - Calibrated parametric medical ultrasound imaging. AB - The goal of this study was to develop a calibrated on-line technique to extract as much diagnostically-relevant information as possible from conventional video format echograms. The final aim is to improve the diagnostic potentials of medical ultrasound. Video-output images were acquired by a frame grabber board incorporated in a multiprocessor workstation. Calibration images were obtained from a stable tissue-mimicking phantom with known acoustic characteristics. Using these images as reference, depth dependence of the gray level could fairly be corrected for the transducer performance characteristics, for the observer dependent equipment settings and for attenuation in the examined tissues. Second order statistical parameters still displayed some nonconsistent depth dependencies. The results obtained with two echoscanners for the same phantom were different; hence, an a posteriori normalization of clinical data with the phantom data is indicated. Prior to processing of clinical echograms,. the anatomical reflections and echoless voids were removed automatically. The final step in the preprocessing concerned the compensation of the overall attenuation in the tissue. A 'sliding window' processing was then applied to a region of interest (ROI) in the 'back-scan converted' images. A number of first and second order statistical texture parameters and acoustical parameters were estimated in each window and assigned to the central pixel. This procedure results in a set of new 'parametric' images of the ROI, which can be inserted in the original echogram (gray value, color) or presented as a color overlay. A clinical example is presented for illustrating the potentials of the developed technique. Depending on the choice of the parameters, four full resolution calibrated parametric images can be calculated and simultaneously displayed within 5 to 20 seconds. In conclusion, an on-line technique has been developed to estimate acoustic and texture parameters with a reduced equipment dependence and to display acoustical and textural information that is present in conventional echograms. PMID- 10823498 TI - Design and optimisation of a simulated moving bed unit: role of deviations from equilibrium theory. AB - The design of a simulated moving bed involves thermodynamic, kinetic and hydrodynamic aspects and requires the optimisation of several variables: plant design variables, such as the column length and diameter, and operating variables, among them four independent flow-rates, the feed concentration and the switch time. In this work we develop an algorithm to design both the unit and its operating conditions, with an overall view on equilibrium properties, efficiency and hydrodynamics, using a simple equilibrium stage model. In this way we determine the parameters leading to the highest possible productivity for a given separation, only requiring the knowledge of the equilibrium isotherms, the Van Deemter equation and a correlation for pressure drop. The algorithm has been used to investigate the effect on the separation performance of some parameters, such as particle size and required product purity, which are not considered by equilibrium theory. The results have been compared with the predictions of equilibrium theory and the observed deviations have been put in evidence and discussed. PMID- 10823499 TI - Complementary mobile-phase optimisation for resolution enhancement in high performance liquid chromatography. AB - An optimisation methodology in high-performance liquid chromatography (HPLC) is presented for the selection of two or more mobile phases having an optimal complementary resolution. The complementary mobile phases (CMPs) are selected in such a way that each one resolves optimally only some compounds in the mixture, while the remainder, resolved by the other mobile phase(s), can overlap among them. The methodology is based on the computation of a peak purity measurement for each solute, using an asymmetrical peak model for peak simulation. Two global resolution criteria (product of elementary resolutions and worst elementary resolution) and two methods for solving the problem (a systematic examination of all possible solute arrangements, and the use of genetic algorithms to expedite the calculation time) were used to find the optimal CMPs. The CMP optimisation methodology was applied to the resolution of a mixture of 10 diuretics and beta blockers, which could not be resolved using a single mobile phase; virtual baseline resolution was achieved, however, with two CMPs. PMID- 10823500 TI - Analysis of linear and cyclic oligomers in polyamide-6 without sample preparation by liquid chromatography using the sandwich injection method. I. Injection procedure and column stability. AB - We report a method for reliable routine polymer sample introduction with minimal bias, a separation method of the first six linear and cyclic oligomers by liquid chromatography, quantification using group equivalents and long term method performance. Injecting a polymer sample in a mobile phase containing an aqueous non-solvent often results in blocked systems as the polymer precipitates in the connecting capillaries. In this first part we focus on a new injection technique, in which the dissolved polyamide is placed between two zones of formic acid, preventing the polymer to precipitate before it reaches the column. Development of this sandwich injection method makes direct injection of the polymer into an aqueous acetonitrile gradient feasible. The oligomeric polyamide recovery of this technique, extraction, dissolution/precipitation and direct injection on a hexafluoro-isopropanol (HFIP) gradient were compared. With the sandwich injection method the polymer remains on the column, slowly changing the stationary phase. The influence of this on resolution and retention was studied. Column stability allows sixty injections before cleaning or replacing the column is necessary. PMID- 10823501 TI - Optimization of ion-exchange displacement separations. I. Validation of an iterative scheme and its use as a methods development tool. AB - Displacement chromatography has been demonstrated to be a powerful, high resolution preparative tool. The performance of displacement systems can be affected by a variety of factors such as the feed load, flow-rate, initial salt concentration and the displacer partition ratio. Thus, the optimization of displacement separations is a uniquely challenging problem. In this manuscript, an iterative optimization scheme has been presented whereby one can identify the optimum operating conditions for displacement separations at a given level of loading on a given resin material. The solid film linear driving force model has been employed in concert with the Steric Mass Action formalism of ion-exchange chromatography to describe the chromatographic behavior in these systems. Simple pulse techniques have been employed to estimate the transport parameters. The iterative scheme has been validated using a rigorous Feasible Sequential Quadratic Programming algorithm. Finally, the utility of the iterative optimization scheme as a methods development tool for displacement separations has been demonstrated for a difficult separation. The results indicate that the use of the optimization scheme leads to significantly better performance than standard rules of thumb. PMID- 10823502 TI - Optimization of ion-exchange displacement separations. II. Comparison of displacement separations on various ion-exchange resins. AB - A variety of stationary-phase materials are currently available for the chromatographic purification of biomolecules. However, the effect of various resin characteristics on the performance of displacement chromatography has not been studied in depth. In Part I, a novel iterative scheme was presented for the rapid optimization of displacement separations in ion-exchange systems. In this article, the optimization scheme is employed to identify the optimum operating conditions for displacement separations on various ion-exchange resin materials. In addition, the effect of different classes of separation problems (e.g., diverging, converging or parallel affinity lines) on the performance of displacement separations is also presented. The solid film linear driving force model is employed in concert with the Steric Mass Action isotherm to describe the chromatographic behavior in these systems. The results presented in this article provide insight into the effects of resin capacity and efficiency as well as the type of separation problem on the performance of various ion-exchange displacement systems. PMID- 10823503 TI - Stereoselective binding of 2,3-substituted 3-hydroxypropionic acids on an immobilised human serum albumin chiral stationary phase: stereochemical characterisation and quantitative structure-retention relationship study. AB - The binding characteristics of a series of 2,3-substituted 3-hydroxypropionic acids, with anti-inflammatory properties, bearing two chiral centres, were studied by HPLC upon HSA (human serum albumin)-based stationary phase. The compounds were analysed in their stereoisomeric erythro and threo forms and the chromatographic conditions for enantioseparation of the erythro and threo forms were studied on human serum albumin stationary phase. The enantiomer elution order was determined by injection of the enriched samples or by carrying out the CD spectra of each enantiomeric fraction. The absolute configuration of the single enantiomers was assigned on the basis of their CD spectra. A QSRR study was performed by subjecting the chromatographic data of the compounds to multiparameter regression analysis against various molecular descriptors to have insight into the chiral recognition mechanism. The lipophilicity appeared to be the most important parameter in determining the affinity to the protein, the compounds' capacity factors being linearly correlated to the experimental RP-HPLC partition coefficients (log k'w). The enantioselectivity factors (alpha) related to the enantiomers of the erythro and threo forms were studied taking into consideration both the physico-chemical parameters and the conformational behaviour of the compounds. PMID- 10823504 TI - Liquid chromatography-electrospray ionization mass spectrometry study of the flavonoids of the roots of Astragalus mongholicus and A. membranaceus. AB - High-performance liquid chromatography-electrospray ionization mass spectrometry has been applied to analyze the flavonoids of Huangqi, the roots of Astragalus mongholicus and A. membranaceus. Eight flavonoids were identified as calycosin-7 O-beta-D-glucoside, calycosin-7-O-beta-D-glucoside-6"-O-malonate (2), ononin, (6aR,11aR)-3-hydroxy-9,10-dimethoxypterocarpan-3-O-bet a-D-glucoside, calycosin, (3R)-7,2'-dihydroxy-3',4'-dimethoxyisoflavan-7-O-beta-D-glucoside, formononetin-7 O-beta-D-glucoside-6"-O-malonate and formononetin by direct comparison with the isolated standards from Huangqi. The existence of (6aR,11aR)-3-hydroxy-9,10 dimethoxypterocarpan, (3R)-7,2'-dihydroxy-3',4'-dimethoxyisoflavan, astrapterocarpanglucoside-6'-O-malonate and astraisoflavanglucoside-6'-O-malonate was detected. This is the first report of flavonoid glycoside malonates in these two Astragalus species, and malonate 2 is a structurally completely identified new compound. PMID- 10823505 TI - Quantitation and characterization of phospholipids in pharmaceutical formulations by liquid chromatography-mass spectrometry. AB - A simple and fast method for phospholipid analysis was developed using high performance liquid chromatography-mass spectrometry with an atmospheric pressure ionization interface. Separation of the phospholipid molecular species was achieved using a linear gradient of a mixture of chloroform-10 mM ammonium acetate-methanol (30:5:65) on a silica column. Optimization of the mass spectrometer conditions has allowed the method to separate and detect the phospholipids mainly as protonated molecular species. In comparison to existing LC-MS methods, improvement in the total analysis time and sensitivity were achieved. Separation of all major phospholipid molecular classes was achieved in less than 6 min. Marked improvement was observed in the linearity of the response of the phospholipids studied providing a linear response over three orders of magnitude. Data supporting the validation of this method for the characterization of major phospholipids molecular species are also presented. PMID- 10823506 TI - Improved liquid chromatographic method for the analysis of photosynthetic pigments of higher plants. AB - The paper presents an improved reversed-phase LC method for the separation of the pigments from green leaves. A good separation of carotenoids and of their cis- and trans-isomers was achieved, especially for the separation of trans-lutein, zeaxanthin, cis-lutein, which are usually not well separated. No perfect separation of alpha-carotene, beta-carotene and pheophytin a was possible, but conditions for a perfect coelution of pheophytin a with either beta-carotene or alpha-carotene were established. Simultaneous equations allowing the determination of pheophytin a and alpha-carotene or pheophytin a and beta carotene are also given. PMID- 10823507 TI - Development and validation of a high-performance liquid chromatographic method using fluorimetric detection for the determination of the diarrhetic shellfish poisoning toxin okadaic acid without chlorinated solvents. AB - A modification of the high-performance liquid chromatographic method with fluorimetric detection method for the determination of diarrhetic shellfish poisoning toxins was developed to completely avoid the use of dangerous chlorinated solvents. The method was validated for the toxin okadaic acid (OA) over a period of 6 months where 12 calibrations were performed and 72 samples were analyzed. Analysis of toxic and non-toxic mussels, clams and scallops demonstrated its selectivity. Linearity was observed in the tested range of interest for monitoring purposes of edible shellfish, from the limit of detection (0.3 microg OA/g hepatopancreas) to 13 microg OA/g hepatopancreas. Intra-assay precision of the method was 7% RSD at the quantification limit (0.97 microg OA/g hepatopancreas at S/N=10). Accuracy was tested in triplicate recovery experiments from OA-spiked shellfish where recovery ranged from 92 to 106% in the concentration range of 0.8 to 3.6 microg OA/g hepatopancreas. Useful information on critical factors affecting calibration and reproducibility is also reported. Good correlation (R=0.87) was observed between the results of the method and those of the method of Lee, after the analysis of 45 samples of mussels from the galician rias. PMID- 10823508 TI - Determination of beryllium in a stream sediment by high-performance chelation ion chromatography. AB - High-performance chelation ion chromatography (HPCIC), involving a chelating silica substrate bonded with aminomethylphosphonic acid, has been developed as a novel technique for the quantitative determination of beryllium in complex matrices. An isocratic separation method, using an eluent containing 1 M KNO3, 0.5 M HNO3 and 0.08 M ascorbic acid, allowed the Be2+ to elute away from the sample matrix peak in under 6 min in a sample containing in excess of 800 mg l( 1) matrix metals. A detection limit of 35 microg l(-1) Be(II) was found using a post-column reaction involving Chrome Azurol S (CAS), 1 M hexamine and 10 mM EDTA buffered at pH 6. The standard addition curve gave excellent linearity (R2>0.999). The procedure was applied to the determination of trace beryllium in a certified sediment sample. The results obtained compared well with the certified value for beryllium. PMID- 10823509 TI - Introduction of large volumes of water-containing samples into a gas chromatograph. Improved retention of volatile solutes through the swing system. AB - The swing system is designed for introducing large volumes of water-containing samples into a gas chromatograph. Sample evaporation and solvent-solute separation are performed in separate compartments. This widens the application range to compounds of higher volatility. Sample evaporation takes place in a hot chamber packed with Carbofrit. Solvent-solute separation is performed in a cascade of increasing powers of retention. While high boiling solutes are retained in an oven-thermostatted retaining precolumn, the more volatile components are retained by a packed bed of sorbents of increasing powers of retention situated in a programmed temperature vaporiser. For elution, the gas flow is reversed and the solutes are discharged from the heated packed bed through the retaining precolumn into the separation column. PMID- 10823510 TI - Gas chromatographic determination of organomercury following aqueous derivatization with sodium tetraethylborate and sodium tetraphenylborate. Comparative study of gas chromatography coupled with atomic fluorescence spectrometry, atomic emission spectrometry and mass spectrometry. AB - Several hyphenated analytical techniques, including gas chromatography (GC) coupled with atomic fluorescence spectrometry (AFS), microwave-induced plasma atomic emission spectrometry (AES), and mass spectrometry (MS), have been evaluated for methylmercury and ethylmercury analysis following aqueous derivatization with both sodium tetraethylborate and sodium tetraphenylborate. Both GC-AFS and GC-AES were shown to be excellent techniques with detection limits in the range of sub-picogram levels (0.02-0.04 pg as Hg). Both techniques have wide linear ranges, although setting of the AFS sensitivity has to be selected manually based on the concentration of mercury in the sample. Phenylation seems to be more favorable in this study because of its capability of distinguishing between ethylmercury and inorganic mercury, and low cost compared to ethylation. Although sensitivity of GC-MS is poor with detection limits ranging from 30 to 50 pg as Hg, it is an essential technique for confirmation of the derivatization products. PMID- 10823511 TI - Simultaneous separation and enantioseparation of thalidomide and its hydroxylated metabolites using high-performance liquid chromatography in common-size columns, capillary liquid chromatography and nonaqueous capillary electrochromatography. AB - The separation of thalidomide (TD) and its hydroxylated metabolites including their simultaneous enantioseparation was studied using three different polysaccharide-type chiral stationary phases (CSPs) in combination with polar organic mobile phases. Three different techniques, high-performance liquid chromatography in common-size columns, capillary LC and nonaqueous capillary electrochromatography were compared in terms of separation. As this study illustrates, polar organic mobile phases represent a valuable extension for less polar and polar aqueous-organic mobile phases in combination with polysaccharide CSPs. Chiralpak AD consisting of 25% of amylose-tris(3,5-dimethylphenylcarbamate) coated on wide-pore aminopropylsilanized silica gel exhibited higher resolving ability compared to the similar cellulose derivative (Chiralcel OD) as well as to cellulose-tris(4-methylbenzoate) (Chiralcel OJ) CSPs for this particular set of chiral analytes. Baseline separation and simultaneous enantioseparation of all three compounds could be achieved under optimized separation conditions. PMID- 10823512 TI - New pseudo-stationary phases for electrokinetic capillary chromatography. Complexes between bovine serum albumin and sodium dodecyl sulfate. AB - The complexes formed between a protein (bovine serum albumin, BSA) and a surfactant (sodium dodecyl sulfate, SDS) were studied as separation carriers in electrokinetic chromatography. Selectivities different from those with either SDS or BSA alone in the background electrolyte (BGE) were obtained. Separation performances were demonstrated to be closely related to the type of complex formed, as predicted by the isotherm curve of SDS on BSA. For each composition of background electrolyte, capacity factors and resolutions were calculated. We compared the results with these complexes to electropherograms using BGE containing either BSA or SDS alone. The separation of a mixture of phenols indicate that some compositions of the BSA-SDS complexes are efficient selectors. PMID- 10823513 TI - Micellar electrokinetic capillary chromatographic separation and fluorescent detection of amino acids derivatized with 4-fluoro-7-nitro-2,1,3-benzoxadiazole. AB - Another method has been developed for the separation of amino acids (1 min derivatization plus 22 min separation) by micellar electrokinetic capillary chromatography (MECC) with laser-induced fluorescence detection. Interestingly enough, such work has never been performed on essential amino acids derivatized by 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F). Fifteen L-amino acid standards were labelled with NBD-F at 60 degrees C for 1 min, and separated in a buffer system containing 20 mM borate, 25 mM sodium cholate, 10 mM Brij 35 and 2.5% methanol. Methanol was employed to expand the MECC migration time window; whereas Brij 35 was used to improve the fluorescence intensity of amino acid derivatives. This method also indicates that bile salt is effective for MECC separation of ionic analytes. Surprising though, improvements in resolution, sensitivity and speed for amino acids analysis are obtained in this work, which are not initially apparent in just employing another derivatizing reagent. Under optimal conditions, 15 amino acids were separated in a short 22 min analysis time, the shortest ever reported, and detection limits of nanomolar concentration and attomole mass were obtained. Furthermore, RSDs of migration time and peak height are better than 1% and 1.8%, respectively, again the smallest ever reported in the literature. PMID- 10823514 TI - Separation of the enzyme cofactor pyrroloquinoline quinone and three isomeric analogues by capillary electrophoresis with ion-pairing media. AB - The enzyme cofactor pyrroloquinoline quinone (PQQ) was successfully separated from three closely related isomeric analogues by capillary electrophoresis with ultraviolet detection. Rapid and efficient separation of all four negatively charged isomers with baseline resolution was achieved by the addition of low concentrations (1-5 mM) of short chain tetraalkylammonium (TAA) salts to the capillary buffer. The TAA cations act as ion-pairing agents and promote differential migration of the isomers with only a minimal reduction in the electroosmotic flow. The effects of the TAA salt concentration and the alkyl chain length were examined. Detection limits of PQQ and its isomers were in the range of 7-15 microM with mass detection limits of 98-210 fmol. PMID- 10823515 TI - Microemulsion electrokinetic chromatography in suppressed electroosmotic flow environment. Separation of fat-soluble vitamins. AB - Microemulsion electrokinetic chromatography (MEEKC) was carried out in a pH 2.5 phosphate buffer to effectively suppress the electroosmotic flow (EOF). With 66.6% (w/w) 25 mM phosphate buffer pH 2.5, 20.0% (w/w) 2-propanol, 6.6% (w/w) 1 butanol, 6.0% (w/w) sodium lauryl sulphate (SDS), and 0.8% (w/w) n-octane as the separation medium, the fat-soluble vitamins A palmitate, E acetate, and D3 were baseline separated within 11 min. With strongly suppressed EOF, the polarity of the separation voltage was reversed (positive electrode at the outlet); the n octane micro droplets surrounded by negatively charged SDS molecules migrated towards the detector. The aqueous part of the microemulsion was modified with 20% (w/w) 2-propanol to improve partition between the n-octane phase and the surrounding aqueous medium. The fat-soluble vitamins were separated in order of decreasing hydrophobicity with a high migration time stability (repeatable within 0.1% RSD). Excellent accuracy and precision were obtained when the system was applied for the determination of vitamin E acetate in commercial vitamin tablets; quantitative data corresponded to 97.0% of label claim, intra-day results varied within 1.72% RSD (n=6), and inter-day results varied within 3.22% RSD (n=5). PMID- 10823516 TI - Development of a capillary electrophoresis method for the simultaneous analysis of artificial sweeteners, preservatives and colours in soft drinks. AB - A rapid capillary electrophoresis method was developed simultaneously to determine artificial sweeteners, preservatives and colours used as additives in carbonated soft drinks. Resolution between all additives occurring together in soft drinks was successfully achieved within a 15-min run-time by employing the micellar electrokinetic chromatography mode with a 20 mM carbonate buffer at pH 9.5 as the aqueous phase and 62 mM sodium dodecyl sulfate as the micellar phase. By using a diode-array detector to monitor the UV-visible range (190-600 nm), the identity of sample components, suggested by migration time, could be confirmed by spectral matching relative to standards. PMID- 10823517 TI - Liquid chromatographic separation of the stereoisomers of thiazide diuretics. AB - A number of racemic thiazide diuretics and analogues were resolved on two diastereomeric chiral stationary phases (CSPs) prepared from (S)- or (R)-alpha-[1 (6,7-dimethyl)naphthyl]-10-dodecenylamine and (S)-2-phenylpropanoic acid. Of the two diastereomeric CSPs, the (S,S) and the (R,S), the former is found to be better than the latter in separating the enantiomers of the racemic thiazide diuretics and their analogues with complete separation being observed on the (S,S)-CSP. Chiral recognition is controlled principally by the (R)- or (S)-alpha [1-(6,7-dimethyl)naphthyl]-10-dodecenylamine portion of the CSPs. The second stereogenic center of the CSP provides but secondary effects on the chiral recognition presumably involving, in the case of the (S,S)-CSP, face-to-edge pi pi interaction between the aromatic ring of the analytes and the phenyl on the second stereogenic center. PMID- 10823518 TI - Ceramide analysis utilizing gas chromatography-mass spectrometry. AB - Suitable analytical methods are a prerequisite of a detailed investigation of ceramides. Therefore, a new gas chromatograph-mass spectrometry method with electron impact ionization was developed. Samples have been prepared for gas chromatography by the formation of volatile trimethylsilyl derivatives. The method provides high separation efficiency, sensitivity and specificity. Mass spectra facilitate the structural characterization of each species, because certain fragments indicate the fatty acid as well as the sphingoid base moiety. In a 30-mm run even very similar ceramides are baseline separated. The method is compared to a recently published assay for liquid chromatography-mass spectrometry. PMID- 10823519 TI - Simultaneous analysis of tea catechins, caffeine, gallic acid, theanine and ascorbic acid by micellar electrokinetic capillary chromatography. AB - A micellar electrokinetic capillary chromatography (MEKC) method for the simultaneous analysis of five tea catechins, theanine, caffeine, gallic acid and ascorbic acid has been developed. The catechins are (-)-epicatechin, (+) catechin, (-)-epigallocatechin, (-)-epicatechin gallate and (-)-epigallocatechin gallate. p-Nitrophenol serves as both reference and internal standard. All the components are separated within 13 min with a 57 cm uncoated fused-silica column. On-column detection was carried out at 200 nm. This method has been used to measure these compounds in fresh tea leaves and tea liquor. The limit of detection for all analytes ranged from 1 to 20 microg/ml. PMID- 10823520 TI - Determination of sunscreen agents in cosmetic products using microwave-assisted extraction and liquid chromatography. AB - Microwave-assisted extraction (MAE), was used to extract sunscreen agents from cosmetic products. The extracts were analyzed by liquid chromatography (LC). The present method allows the determination of three sunscreen agents, Eusolex 2292, 4360 and 6300. The precision of the assay at 40 microg/ml of sunscreen agents ranged from 1.5 to 2.2%, and the detection limits were 2.0-4.0 ng/ml. PMID- 10823521 TI - Whole blood D-dimer assay: an effective noninvasive method to rule out pulmonary embolism. AB - BACKGROUND: The whole blood D-dimer assay has gained recognition as a noninvasive test to rule out pulmonary embolism (PE) in medical patients. METHODS: We performed a whole blood D-dimer assay in medical and surgical patients undergoing either pulmonary angiogram or pulmonary ventilation perfusion scan for suspected PE or duplex Doppler or venogram for suspected deep venous thrombosis (DVT). RESULTS: A total of 483 patients were enrolled; 16 were excluded because of an equivocal pulmonary ventilation perfusion scan. The 467 remaining patients had a mean age of 56 +/- 27 years. There were 258 women and 209 men. A total of 353 patients were admitted to a medical service and 114 to surgery/ trauma. A total of 82 patients (18%) developed thromboembolism: 20 had PE, and 62 had DVT. CONCLUSION: No surgical patient with PE or DVT (n = 27) had a negative D-dimer. A negative D-dimer result in a stable surgical patient should be considered conclusive evidence to rule out thromboembolism and, thus, negate the need for further diagnostic studies. In our surgical patients suspected of DVT or PE, had D-dimer been used, one third of the patients would have avoided an expensive or invasive diagnostic test. PMID- 10823522 TI - Blunt splenic injuries: dedicated trauma surgeons can achieve a high rate of nonoperative success in patients of all ages. AB - BACKGROUND: Selective nonoperative management (NOM) of blunt splenic injuries is becoming a more prevalent practice. Inclusion criteria for NOM, which have been a source of controversy, continue to evolve. Age > or = 55 years has been proposed as a predictor for failure of and even a contraindication to NOM of blunt splenic trauma. Additionally, the high rate of NOM in children (up to 79%) has been attributed to their management by pediatric surgeons. We evaluated our experience with NOM of blunt splenic injury with special attention to these age groups. METHODS: By using our trauma registry, all patients with blunt splenic injuries (documented by computed tomography, operative findings, or both) cared for over a 36-month period, at a single American College of Surgeons verified Level I trauma center were reviewed. Detailed chart reviews were performed to examine admission demographics, laboratory data, radiologic findings, outcome measures, and patient management strategy. All patients were managed by nonpediatric trauma surgeons. We then compared our adult data with that in the recent literature and our pediatric data with that of the National Pediatric Trauma Registry over the same time period. RESULTS: We identified 251 consecutive patients with blunt splenic injuries. Eighteen patients who expired in the immediate postinjury period were excluded from statistical evaluation. No deaths occurred as a result of splenic injury. Of the remaining 233 patients, 73 patients (31%) required early celiotomy, 160 patients (69%) were selected for NOM, with 151 patients (94%) being successfully managed without operation. Blunt splenic injury occurred in 23 patients age 55 years or older. Eighteen patients (78%) were selected for NOM and 17 patients (94%) were successfully treated without operation. Blunt splenic injury occurred in 35 patients less than 16 years of age. Thirty-two patients (91%) were selected for NOM. Thirty-one patients (89% of all pediatric patients) were successfully treated without operation. CONCLUSION: Age > or = 55 years is not a contraindication to nonoperative management of blunt splenic injuries. Children with blunt splenic injuries can be successfully managed nonoperatively by nonpediatric trauma surgeons. PMID- 10823523 TI - Septic mucosal intraepithelial lymphoid immune suppression: role for nitric oxide not interleukin-10 or transforming growth factor-beta. AB - OBJECTIVE: Recent studies indicate that sepsis induces a marked depression in the splenocyte immune response (as illustrated by decreased interleukin [IL]-2 production, interferon [IFN]-gamma production, or both) in response to T-cell mitogen. However, it is not known whether a similar depression is evident in the phenotypically distinct, small intestine intraepithelial lymphocytes (IELs) or what regulates this process during sepsis. Because the maintenance of a competent mucosal immune response is thought to be central to the animal's ability to survive sepsis, we attempted to determine whether IEL's IL-2/IFN-gamma production is suppressed and what mediates this depression. RESULTS: Our studies indicated that C3H/HeN mice subjected to cecal ligation and puncture (CLP) exhibited a marked decline in the ability of IELs to release IL-2/IFN-gamma at 24 hours and that this decline is associated with increased secretion of IL-10 and nitric oxide (NO). To the extent that IL-10 accounted for this loss of IL-2/IFN-gamma release, we observed that IL-10 gene deficiency neither restored the IL-2/IFN gamma release nor suppressed the increase in NO when compared with background control, C57BL/6J mouse cells. To further study whether NO was involved in this immune suppression, iNOS knockout (iNOS -/-) were also subjected to the same procedure; however, the depression in IL-2/IFN-gamma was not seen in iNOS -/- mice when compared with background controls. CONCLUSION: Our data indicate that IL-10, which affects splenic lymphoid response, may not be a key mediator of IEL immune suppression and that the induction of NO may play a more significant role in gastrointestinal immune dysfunction seen in late sepsis. PMID- 10823524 TI - Difference in the responses after administration of granulocyte colony stimulating factor in septic patients with relative neutropenia. AB - OBJECTIVE: The objective of this study was to classify the clinical responses after administration of granulocyte colony-stimulating factor (G-CSF) in septic patients with relative neutropenia. PATIENTS AND METHODS: We administered recombinant human G-CSF (2 microg/kg) subcutaneously once a day for 5 days to 30 septic patients with white cell counts below 5,000 cells/mm3. Absolute neutrophil count (ANC), neutrophil differentiation, and serum concentration of G-CSF were determined serially. Bone marrow also was analyzed before and after treatment. RESULTS: Neutrophil responses to G-CSF varied from good (ANC > 10,000/mm3, group G, n = 20) to moderate (ANC < 10,000/mm3, group M, n = 5) to poor (no increase in ANC, group P, n = 5). Before G-CSF administration, the three groups showed no differences in ANC but did show significant differences in serum concentration of G-CSF. G-CSF concentration was 0.16 +/- 0.03 ng/mL in group G, 7.0 +/- 3.0 ng/mL in group M, and 270 +/- 90 ng/mL in group P. Immature neutrophils accounted for 35.0 +/- 3.7% of peripheral leukocytes in group P but only 5.1 +/- 0.6% in group G. Although bone marrow was depressed in all groups before G-CSF treatment, nucleated cell count increased significantly after rhG-CSF treatment in groups G and M. Survival rate after 4 weeks was 90% in group G and 100% in group M; no patient in group P survived. CONCLUSION: G-CSF administration was effective in septic patients with a low percentage of immature neutrophils and insufficient endogenous G-CSF. It had little effect on patients with a high percentage of immature neutrophils whose G-CSF production was up-regulated and whose bone marrow was severely depressed. PMID- 10823525 TI - Prostaglandin E2 receptor antagonist (SC-19220) treatment restores the balance to bone marrow myelopoiesis after burn sepsis. AB - BACKGROUND: Although prostaglandin E2 (PGE2) has been shown to be immunosuppressive, its role in the development of specific bone marrow myeloid lineages after thermal injury and sepsis has yet to be elucidated. The purpose of this study was to demonstrate that alterations in bone marrow progenitor proliferation favoring monocytopoiesis in burn sepsis can be restored by blocking the cellular interactions of PGE2. METHODS: A murine model of burn sepsis with and without treatment with SC-19220, a PGE2 receptor antagonist, was used to determine peripheral monocyte and neutrophil counts as well as the colony forming potential of colony-stimulating factor responsive bone marrow progenitors. RESULTS: Burn sepsis augmented the growth of the early colony-forming unit granulocyte-macrophage and monocyte progenitors and the number of circulating monocytes, whereas granulocyte progenitors and circulating neutrophils demonstrated an opposite response. Treatment with SC-19220 nearly reversed these alterations. CONCLUSION: These data indicate that abrogating PGE2's actions during burn sepsis can restore the balance in bone marrow granulocyte and monocyte production, further consolidating the pivotal role PGE2 plays in the pathogenesis of burn sepsis. PMID- 10823526 TI - Sex differences in posttraumatic cytokine release of endotoxin-stimulated whole blood: relationship to the development of severe sepsis. AB - BACKGROUND: In experimental trauma-hemorrhage and sepsis, a sexual dimorphism of cell-mediated immune functions has been described, which has been related to higher susceptibility to and mortality from sepsis in males. Therefore, in the present study, sex differences with regard to cytokine release of endotoxin stimulated whole blood and its relation to the development of severe posttraumatic sepsis were investigated in blunt trauma patients with multiple injuries. METHODS: Eighty-four patients (25 female; 59 male) sustaining blunt injuries with an Injury Severity Score > 16 were enrolled in the study. Whole blood and serum were obtained during a 14-day period of hospitalization. The capacity of peripheral blood mononuclear cells to produce cytokines (tumor necrosis factor-alpha, interleukin [IL]-6, IL-8) was tested by using a whole blood assay. Serum samples were assayed for anti-inflammatory cytokines (IL-4, IL 10, and transforming growth factor beta1) and sex hormones (testosterone, estradiol, progesterone). Patients were monitored daily for sepsis criteria according to the ACCP/ SCCM consensus conference 1992. RESULTS: Within the entire patient population, sex differences in posttraumatic cytokine release were not detectable. Male trauma patients developing severe sepsis (n = 16) presented with a significantly increased cytokine producing capacity in the early posttraumatic period (< or = 24 hours after admission to the emergency room) when compared with males with an uncomplicated recovery. In females, differences between the subgroups of patients with (n = 7) and without development of severe sepsis were not detectable. There were no differences in systemic levels of anti-inflammatory cytokines within the early posttraumatic period between the subgroups of male and female patients with and without development of severe sepsis. In females, differences in sex hormone levels were not detectable, whereas in males, development of severe sepsis later was found to coincide with significantly decreased testosterone and increased estradiol serum levels. CONCLUSION: The present study demonstrates a sex-specific regulation of leukocyte function in patients with multiple injuries within the early posttraumatic period. In male patients with multiple injuries, increased cytokine-producing capacities may correspond to enhanced inflammatory responses, which increase susceptibility to sepsis, whereas in female patients, other regulatory mechanisms may be involved. PMID- 10823527 TI - Outcome from injury: general health, work status, and satisfaction 12 months after trauma. AB - OBJECTIVE: We evaluated outcomes 12 months after trauma in terms of general health, satisfaction, and work status. METHODS: Two hundred forty-seven patients without severe neurotrauma were evaluated by interview during admission and by mailed self-report 6 and 12 months after trauma. Data were obtained from the Trauma Registry, interviews, and survey instruments. Baseline assessment was obtained with the Short Form 36 (SF36) and the Sickness Impact Profile (SIP) work scale. Outcome measures were the SF36, SIP work scale, Brief Symptom Inventory (BSI) depression scale, the Civilian Mississippi Scale for Posttraumatic Stress Disorder (PTSD), and a satisfaction questionnaire. Three regressions were determined for outcome. The dependent variables were general health and work status (linear) and satisfaction (logistic). Each regression controlled for baseline status and mental health, Injury Severity Score (ISS), and 12-month SF36 physical function before evaluating the effect of outcome mental health. RESULTS: Follow-up data were available for 75% of the patients at 6 months and 51% at 12 months. The mean age of patients was 37.2 +/- 0.9 years (+/-SEM), and 73% were male. Their average ISS was 13.9 +/- 0.6. Seventy percent of injuries were blunt force, 13.5 % were penetrating, and 16.5 % were burn injuries (mean total body surface area, 13.3 +/- 1.5%). Sixty-four percent of the patients had returned to work at 12 months. Follow-up SF36 mental health was associated with the dependent outcome in each regression. After controlling for baseline status and mental health, ISS, and outcome SF36 physical function, outcome mental health was associated with outcome SF36 general health (p < 0.001), SIP work status (p = 0.017), and satisfaction with recovery (p = 0.005). Outcome SF36 mental health was related to baseline mental health, 12-month PTSD and BSI depression scores, and increased drug and alcohol use. CONCLUSIONS: Twelve months after trauma, patients' work status, general health, and overall satisfaction with recovery are dependent on outcome mental health. This dependency persists despite measured baseline status, ISS, or physical recovery. The mental disease after trauma is attributable to poor mental health, the development of symptoms of PTSD and depression, and increased substance abuse. Trauma centers that fail to recognize, assess, and treat these injury-related mental health outcomes are not fully assisting their patients to return to optimal function. PMID- 10823528 TI - Prehospital resuscitation with phenylephrine in uncontrolled hemorrhagic shock and brain injury. AB - BACKGROUND: Hypotension doubles the adverse outcome of severe brain injury (BI). This finding is thought to be due to secondary ischemia caused by cerebral hypoperfusion. Aggressive prehospital fluid resuscitation in BI is advocated to maintain mean arterial pressure (MAP). Increasing MAP by prehospital fluid resuscitation before control of hemorrhage is thought to increase blood loss and reduce survival. We hypothesized that vasoconstrictor treatment of uncontrolled hemorrhage would increase MAP, reduce hemorrhage volume, and decrease the extent of BI compared with delayed fluid resuscitation (DR) or resuscitation with Ringer's lactate (RL). METHODS: Swine were randomly assigned to a control group or an experimental group: splenic laceration (uncontrolled hemorrhage) and cryogenic BI. The experimental group received one of three prehospital resuscitation regimens: DR, RL, or phenylephrine (Phen) to maintain baseline MAP. Variables were measured at baseline and at 20, 50, and 120 minutes during the simulated "prehospital and early hospital" phases and at 2 and 8 hours after surgical control of the uncontrolled hemorrhage. After killing, biopsies of the brain, liver, kidney, and gut were evaluated for histologic evidence of ischemia and compared between groups. RESULTS: Hemorrhage volume was similar in the experimental groups. Mortality was lowest in the Phen group (11%) compared with DR (40%) and RL (33%) groups. Phen increased MAP and cerebral perfusion pressure. RL infusion increased cerebral blood flow and resulted in less secondary injury than either Phen or DR. CONCLUSION: Phen improves MAP and systemic and cerebral perfusion pressure in the prehospital phase but does not reduce secondary neuronal ischemia. RL restores cerebral blood flow earlier and is associated with less secondary ischemia than either Phen or DR in this model. These data suggest that prehospital infusion of RL in patients with BI and shock is warranted and decreases secondary ischemia. PMID- 10823529 TI - Giving bad news: the family perspective. AB - BACKGROUND: Death from trauma frequently comes without forewarning. Relating the news of death to the family is often the responsibility of trauma surgeons. The purpose of this study was to investigate the key characteristics and methods of delivering bad news from the perspective of surviving family members. METHODS: We designed and administered a survey tool to surviving family members of trauma patients dying in the emergency department or intensive care unit. The tool consisted of 14 elements that surviving family members graded in importance when receiving bad news (1, least; 6, most). Respondents also judged the attention given to these elements (good, fair, or poor) by the person giving the bad news of death. RESULTS: Fifty-four family members of 48 patients who died completed the survey (44 intensive care unit deaths, 4 emergency room deaths). Deceased patients ranged in age from 12 to 91 years (mean, 53 years). Death occurred within 2 days of injury in 69% of the patients and within 1 week in 83%. The most important features of delivering bad news were judged to be attitude of the news giver (ranked most important by 72%), clarity of the message (70%), privacy (65%), and knowledge/ ability to answer questions (57%). The attire of the news giver ranked as least important (3%). Sympathy, time for questions, and location of the conversation were ranked of intermediate importance. Touching was unwanted by 30% of the respondents, but encouraged or acceptable in 24%. CONCLUSION: The attitude of the news-giver, combined with clarity of the message and the time, privacy, and knowledge to answer questions are the most important aspects of giving bad news. This information should be incorporated into resident training. PMID- 10823530 TI - Effects of increased renal parenchymal pressure on renal function. AB - OBJECTIVE: Acute renal failure is seen with the acute abdominal compartment syndrome (AACS). The cause of acute renal failure in AACS is thought to be multifactorial, including increased renal venous pressure, renal parenchymal pressure (RPP), and decreased cardiac output. Previous studies have established the role of renal venous pressure as an important mediator of this renal derangement. In this study, we evaluate the role of renal parenchymal compression on renal function. METHODS: Two groups of swine (20-26 kg) were studied after left nephrectomy and placement of a renal artery flow probe and ureteral cannula. Two hours were allowed for equilibration, and an inulin infusion was begun to calculate inulin clearance as a measurement of glomerular filtration. In group 1 animals (n = 6), RPP was elevated by 30 mm Hg for 2 hours with renal parenchymal compression. RPP then returned to baseline for 1 hour. In group 2 (n = 6), the RPP was not elevated. The cardiac index, preload, and mean arterial pressure remained stable. Blood samples for plasma renin activity and plasma aldosterone were taken at baseline and at hourly intervals. RESULTS: Elevation of RPP in the experimental group showed no significant decrease in renal blood flow index or glomerular filtration when compared with control animals. There were no significant elevations of plasma aldosterone or plasma renin activity in the experimental animals when compared with control. CONCLUSION: Elevated renal compression alone did not create the pathophysiologic derangements seen in AACS. However, prior data from this laboratory found that renal vein compression alone caused a decreased renal blood flow and glomerular filtration and an increased plasma renin activity, plasma aldosterone, and urinary protein leak. These changes are partially or completely reversed by decreasing renal venous pressure as occurs with abdominal decompression for AACS. These data strengthen the proposal that renal vein compression, and not renal parenchymal compression, is the primary mediator of the renal derangements seen in AACS. PMID- 10823531 TI - Effects of fiberoptic bronchoscopy on intracranial pressure in patients with brain injury: a prospective clinical study. AB - BACKGROUND: Fiberoptic bronchoscopy (FB) plays an important role in making the diagnosis of nosocomial pneumonia and resolving lobar atelectasis in critically injured trauma patients. It has been shown to be a safe procedure with only occasional complications. However, in patients with head injuries, FB can lead to intracranial hypertension. Sustained increases in intracranial pressure (ICP) leads to poor outcome in these patients. Because of this, a prospective study was done not only to assess the effect of FB on ICP and cerebral perfusion pressure (CPP) in patients with brain injuries, but also to identify a regimen of sedation and anesthesia that could prevent significant increases in ICP during FB. METHODS: Twenty-six FB were performed in 23 patients with ICP monitors or ICP monitors and ventriculostomy drains in place for Glasgow Coma Scale score < 8 or management of postcraniotomy trauma. FB was performed to aid in the diagnosis of nosocomial pneumonia or to aid in resolving lobar atelectasis. Before FB, all patients received a standard anesthetic regimen consisting of vecuronium (10 mg), morphine sulfate (4 mg), and midazolam (2.5 mg). Patients with diminished cranial compliance, defined as ICP > 10 mm Hg, also received a nebulizer treatment of 3 mL of 4% lidocaine before FB. All patients were preoxygenated with FIO2 = 1.0 for 10 minutes. Intracranial pressure, mean arterial pressure, and CPP were monitored continuously throughout the procedure. These same variables were also recorded at baseline and at 2-minute intervals during the procedure. The time to return to baseline ICP was also recorded. RESULTS: The mean ICP at baseline (immediately before FB) was 12.6 mm Hg. After introduction of the bronchoscope, the ICP rapidly increased in 21 procedures (81%) and the mean highest ICP was 38.0 mm Hg. There was also a concomitant increase in mean arterial pressure such that there was no substantial change in CPP. The mean lowest CPP was 73.1 mm Hg. The average time for return of ICP to baseline was 13.9 minutes. In the subgroup of patients with ICP > 10, attempting to blunt the tracheal stimulation by anesthetizing the trachea with 4% nebulized lidocaine did not seem to be successful. The mean highest ICP in this subgroup was 41.8 mm Hg. The CPP changed in a similar manner, as the mean lowest CPP was 74.0 mm Hg. The mean time to return to baseline was 12.5 minutes. No patient had acute neurologic deterioration secondary to FB. CONCLUSIONS: Although FB is an important procedure in the pulmonary care of head injured patients, it produces substantial, but transient, increases in ICP and should be used with caution in patients with diminished cranial compliance. Sedation, analgesia, paralysis, and topical tracheal anesthesia did not completely prevent the rise in ICP. Although no acute deterioration in condition occurred, secondary brain injury caused by localized cerebral ischemia is certainly possible. Because of the substantial increases in ICP, herniation may be precipitated in an occasional patient. Further study is needed to identify a regimen that will confer protection. PMID- 10823532 TI - Comparison of various hemoglobin polyoxyethylene conjugate solutions as resuscitative fluids after hemorrhagic shock. AB - BACKGROUND: Previous research suggested that splanchnic hypoperfusion occurs after resuscitation with certain acellular hemoglobin solutions. We examined the influence of maltose content and oxygen affinity on resuscitation with various hemoglobin polyoxyethylene conjugate solutions after hemorrhage. METHODS: Fifteen swine underwent hemorrhage and equal volume resuscitation with pyridoxalated hemoglobin polyoxyethylene conjugate containing 0% or 8% maltose, or low P50 conjugate, which also contained 8% maltose. Five control animals were monitored but not bled. Regional blood flow was determined by using radioactive microspheres, gastric mucosal perfusion was estimated with tonometry, and gut histopathology was evaluated. RESULTS: All hemoglobin solutions produced vasoconstriction, manifested by elevated mean systemic and pulmonary artery pressures without a significant decrease in cardiac index compared with the sham group. Resuscitation with maltose-containing solutions elevated arterial and regional PCO2 and depressed arterial pH and gastric pHi (p < 0.05 for all). Splanchnic and renal blood flows were reduced in the low P50 + 8% maltose group (p < 0.05 vs. sham and baseline for renal blood flow), possibly indicating greater regional vasoconstriction in this group. Ileal mucosal damage was more severe in the maltose-containing groups and correlated with decreased pHi. CONCLUSION: Vasoconstriction occurred in all groups but was more severe in the low P50 + 8% maltose group. Maltose-containing solutions caused respiratory acidosis, decreased pHi, and histologic evidence of mucosal injury. Pyridoxalated hemoglobin polyoxyethylene conjugate without maltose was a superior resuscitation solution in this swine model. PMID- 10823533 TI - Operative management of injured children at an adult level I trauma center. AB - BACKGROUND: The geographic distribution of trauma centers results in a significant number of children being treated in adult centers. The emphasis on nonoperative management of pediatric blunt trauma has heightened concern that in adult trauma centers, an aggressive operative approach will continue to be used. We hypothesized that pediatric commitment at a Level I trauma center results in appropriate nonoperative care of injured children as established by regional pediatric trauma centers. METHODS: The records of 1,792 consecutively treated children admitted to the trauma service during a 6-year period (January of 1990 to December of 1995) were reviewed. Patients were stratified into one of three age groups: 0 to 5, 6 to 11, and 12 to 17 years of age. RESULTS: Mean age of the study patients was 10.0 +/- 0.1 years, 1,147 were boys (64%), and their mean Injury Severity Score was 7.3 +/- 0.3. The injury mechanism was blunt in 1,550 (87%) and 132 (7%) required laparotomy. In the 0- to 5-year-old blunt mechanism group, 6% underwent laparotomy or thoracotomy from 1990 to 1992. In comparison, only 1% of this age group had a laparotomy from 1993 to 1995 (p < 0.05, Fisher's exact test). A similar trend was found in the 6- to 11-year-old children after blunt injury (4% laparotomy rate from 1990 to 1992; 2% from 1993-1995). CONCLUSION: There has been a declining trend in the operative management of blunt pediatric trauma, especially in children less than 6 years old, whereas the operative management of penetrating injuries has remained stable. These data confirm that pediatric commitment in a Level I trauma center results in nonoperative treatment of injured children commensurate with that established in regional pediatric trauma centers. PMID- 10823534 TI - Focused abdominal sonography for trauma (FAST) in children with blunt abdominal trauma. AB - BACKGROUND: Focused abdominal sonography for trauma (FAST) has been well reported in adults, but its applicability in children is less well established. We decided to test the hypothesis that FAST and computed tomography (CT) are equivalent imaging studies in the setting of pediatric blunt abdominal trauma. METHODS: One hundred seven hemodynamically stable children undergoing CT for blunt abdominal trauma were prospectively investigated using FAST. The ability of FAST to predict injury by detecting free intraperitoneal fluid was compared with CT as the imaging standard. RESULTS: Thirty-two patients had CT documented injuries. There were no late injuries missed by CT. FAST detected free fluid in 12 patients. Ten patients had solid organ injury but no free fluid and, thus, were not detected by FAST. The sensitivity of FAST relative to CT was only 0.55 and the negative predictive value was only 0.50. CONCLUSION: FAST has insufficient sensitivity and negative predictive value to be used as a screening imaging test in hemodynamically stable children with blunt abdominal trauma. PMID- 10823535 TI - Upper extremity fractures in restrained front-seat occupants. AB - BACKGROUND: The aim of our study was to analyze the injury mechanism of upper extremity fractures in car crashes, to create a basis for developing prophylactic devices. METHODS AND RESULTS: During 1985-1995, 3,260 restrained car drivers (1,228 front-seat passengers) were injured in 9,380 crashes involving cars. A total of 179 drivers (5.5%) (front-seat passengers: n = 53, 4.3%) sustained fractures of the arm. The hand (25%), wrist (23%), and forearm (23%) were affected most often, and the elbow (9%), upper arm (10%), and shoulder (10%) were seldom affected. No considerable differences of the injury mechanism were found comparing drivers with front-seat passengers. Fractures were mainly caused by head-on collisions (n = 119, 51%) or multiple collisions (n = 78, 34%). In 73% of the crashes (n = 166), delta-v exceeded 30 km/h (18.6 mph). A lower Delta-v resulted mainly in fractures affecting the shoulder and wrist. CONCLUSION: Because more than half of the upper extremity fractures resulted from a direct impact to the hand, arm, or both, modifications to improve the energy absorption by padding dashboard and inner door or by additional airbags are promising. PMID- 10823536 TI - Utility of both muscle and fascia flaps in severe lower extremity trauma. AB - BACKGROUND: The evolving technology in trauma management today permits salvage of many severe lower extremity injuries previously even considered to be lethal. An essential component for any such treatment protocol must be adequate soft tissue coverage that often will use vascularized flaps. Traditionally, calf muscles have been used proximally and free flaps for the distal leg and foot. The reintroduction of reliable local fascia flaps has challenged this dictum, proving to be a simpler and yet versatile option. MATERIALS AND METHOD: The role of both muscle and fascia flaps in lower extremity injuries has been retrospectively reviewed from a 2-decade experience. Soft tissue deficits requiring some form of vascularized flap occurred in 160 limbs in 155 patients. The frequency of use of flap types, specific complications and benefits, effect of timing of wound closure, and rate of limb salvage were compared. RESULTS: Initial coverage after significant lower extremity trauma in these 160 limbs required 60 local muscle flaps, 50 local fascia flaps, and 74 free flaps. These flaps had been selected on a nonrandom basis according to wound location, its severity, and flap availability. Complications were directly related to the severity of injury, and for free flaps as a group (39%), although these were not independent variables. Local muscle (27%) or fascia flaps (30%) were similar with regard to this morbidity. Healing was more likely to be uneventful if coverage were accomplished during the acute period after injury, regardless of flap type. Muscle flaps were still used in two thirds of all cases, with the soleus muscle used as often for the distal leg as the mid-leg. Local fascia flaps were most valuable for smaller defects, especially in the distal leg or foot, and often as a reasonable alternative to a free flap. CONCLUSION: The traditional role of the gastrocnemius muscles for flap coverage of knee and proximal leg defects and the soleus muscle for the middle third of the leg was reaffirmed. The soleus muscle often also reached distal leg defects as could local fascia flaps, where classically, otherwise, a free flap would have been necessary. The largest or most severe wounds, irrespective of limb location, required free flap coverage. Local fascia flaps proved to be a valuable alternative. PMID- 10823537 TI - Association of endotoxemia and production of antibodies against endotoxins after multiple injuries. AB - BACKGROUND: Endotoxemia after injury has been a controversial issue. Endotoxins stimulate the innate and adaptive immune system. OBJECTIVE: To investigate endotoxemia and its effects on the production of antiendotoxin antibodies of cultured mononuclear cells of patients with multiple injuries. METHODS: Blood samples of 20 patients with multiple injuries were collected up to 12 days after trauma. The endotoxin concentration was measured in the plasma, and mononuclear cells were isolated and cultured. Specific antibodies against two lipopolysaccharides, one lipid A preparation, and alpha-hemolysin of Staphylococcus aureus were measured in the cell culture supernatant by an enzyme linked immunosorbent assay. RESULTS: Endotoxemia peaked at admission of the patients, decreasing thereafter to almost normal values within 5 days. Isolated mononuclear cells synthesized antibodies against all tested antigens with a peak at or between day 5 and day 7. The increase was significant for immunoglobulin (Ig)A and IgM specific to all endotoxins tested and for IgA specific to alpha hemolysin. However, there were no significant changes of the concentrations of total IgM, IgA, and IgG. All specific IgG remained unaffected. CONCLUSION: Patients with multiple injuries initially have temporary endotoxemia. Endotoxin may be suggested as a stimulator of the synthesis of antiendotoxin antibodies, in particular of the IgA and IgM class in patients with multiple injuries. PMID- 10823538 TI - Evaluation of incidence, clinical significance, and prognostic value of circulating cardiac troponin I and T elevation in hemodynamically stable patients with suspected myocardial contusion after blunt chest trauma. AB - BACKGROUND: The frequency and prognostic influence of myocardial injury in patients with blunt chest trauma is controversial. We investigated the value of cardiac troponin I (cTn-I) and cardiac troponin T (cTn-T), highly specific markers of myocardial injury, to determine whether their measurement would improve the ability to detect myocardial contusion in stable patients with blunt chest trauma in comparison with conventional markers and whether they were associated with significantly worse late clinical outcome. METHODS AND RESULTS: Over an 18-month period, myocardial contusion was diagnosed in 26 of 94 patients (27.6%) with acute blunt chest trauma (motor vehicle crash; 81%), because of echocardiographic abnormalities (n = 12), electrocardiographic abnormalities (n = 29), or both. Patients with myocardial contusion had a significantly higher Injury Severity Score at the time of admission (p = 0.001) and a significantly longer hospital stay (p = 0.0008). All patients survived admission to hospital and were hemodynamically stable. None of the patients died or had severe in hospital cardiac complications. The percentage of patients with elevated CK, (CK MB/total CK) ratio, or CK-MB mass concentration was not significantly different between patients with or without myocardial contusion. However, there were significant differences between the two groups when we applied the commonly used threshold levels of CK-MB activity and myoglobin. The percentage of patients with elevated circulating cTn-I and cTn-T (> or = 0.1 microg/L) was significantly higher in patients with myocardial contusion (23% vs. 3%; p = 0.01 and 12% vs. 0%; p = 0.03, respectively). Complete changes in cTn-I and cTn-T correlated well (r = 0.91, p = 0.0001). Sensitivity, specificity, and negative and positive predictive values of cTn-I and cTn-T in predicting a myocardial contusion in blunt trauma patients were 23%, 97%, and 77%, 75%, and 12%, 100%, and 74%, 100%, respectively. Clinical follow-up was available in 83 patients (88%) (mean, 16 +/- 7.5 months). There were no deaths in either group directly attributed to cardiac complications. None of the patients had any long-term cardiac complications or myocardial failure related to blunt chest trauma. CONCLUSION: Although improved specificity of cTn-I and cTn-T compared with conventional markers, it should be emphasized that the main problem with cTn-I and cTn-T is low sensitivity as well as low predictive values in diagnosing myocardial contusion. cTn-I and cTn-T measurement is currently not an improved method in diagnosing blunt cardiac injury in hemodynamically stable patients. Moreover, there was no association of postmyocardial contusion cell injury and late outcome in these patients when cTn I and cTn-T and other conventional markers were considered. PMID- 10823539 TI - Incidence of septic complications and multiple organ failure in severely injured patients is sex specific. AB - BACKGROUND: Sexual hormones are potent regulators of various immune functions. Although androgens are immunosuppressive, estrogens protect against septic challenges in animal models. This study correlates sexual dimorphism with the incidence of posttraumatic complications in severely injured patients. METHODS: From January of 1991 to February of 1996, 1,276 consecutive injured patients (Injury Severity Score [ISS] > or = 9 points) were studied. Males (n = 911) did not differ from females (n = 365) with regard to severity of injury (ISS) and injury pattern. RESULTS: The incidence of posttraumatic sepsis (30.7%) and multiple organ dysfunction syndrome (29.6%) was significantly increased in severely injured males with ISS > or = 25 points in comparison to the equivalent group of females (sepsis, 17.0%; multiple organ dysfunction syndrome, 16.0%). No difference was found in patients with ISS < 25 points. Moreover, plasma levels of procalcitonin and interleukin-6 were elevated (p < 0.05) in severely injured males compared with females. CONCLUSION: Sex influences posttraumatic morbidity in severely injured patients and supports the concept that females are immunologically better positioned toward a septic challenge. PMID- 10823540 TI - Comparison of impact data in hockey, football, and soccer. AB - PURPOSE: To compare accelerational forces to the head in high school-level football, hockey, and soccer athletes. METHODS: Acceleration of impact was measured within the helmet of high school hockey and football players during actual game play. A triaxial accelerometer was placed at the vertex of the helmet immediately adjacent to the players head. Peak acceleration (in g's) was measured and the Gadd Severity Index and Head Injury Criterion score calculated during actual play periods in several games over four seasons. We also recorded acceleration of head impacts in high school-level soccer players who headed a soccer ball while equipped with a football helmet instrumented identically to the helmet used to record during football games. RESULTS: Peak accelerations inside the helmet for football averaged 29.2 g compared with 35 g for hockey (p = .004). There were no incidents of concussion or other traumatic brain injury during the recorded periods. In contrast, the peak accelerations associated with heading a soccer ball was 54.7 g (p = 2 x 10(-5) vs. hockey). CONCLUSION: Peak accelerations as measured at the surface of the head were 160 to 180% greater from heading a soccer ball than from routine (noninjurious) impacts during hockey or football, respectively. The effect of cumulative impacts at this level may lead to neurologic sequelae. PMID- 10823541 TI - Surface cooling, which fails to reduce the core temperature rapidly, hastens death during severe hemorrhagic shock in pigs. AB - OBJECTIVE: To examine whether surface cooling (SC) would rapidly decrease the core temperatures and prolong the survival time during volume-controlled lethal hemorrhagic shock in pigs. METHOD: Twelve pigs were randomly assigned to the SC group (group 1, n = 6) or the no cooling control group (group 2, n = 6), after blood withdrawal of 30 mL/kg over 15 minutes, and maintained under spontaneous breathing by light anesthesia with 1.0% halothane. SC was performed by applying ethanol to the skin, blowing with an electric fan, and placing ice packs. Pigs were observed without fluid resuscitation until their death (apnea and no pulse). RESULTS: SC did not lower the rectal temperature (Tr) to 35 degrees C at any time point until death, except one pig; in that animal, Tr was decreased to 34 degrees C after 135 minutes from the start of SC. The survival time was 108 +/- 43 minutes in group 1 and 175 +/- 55 minutes in group 2 (p < 0.05, life table analysis). CONCLUSION: In lightly anesthetized pigs during hemorrhagic shock, SC without resuscitation did not rapidly reduce the core temperature and rather hastened death for reasons that remain to be explored. PMID- 10823542 TI - Endovascular stent-graft placement for treatment of traumatic penetrating subclavian artery injury. PMID- 10823543 TI - Axillary artery injury: an isolated, delayed case after blunt trauma of the upper limb. PMID- 10823545 TI - Iatrogenic arterial trauma associated with hip fracture treatment. PMID- 10823544 TI - Transjugular intrahepatic portosystemic shunt for trauma? AB - We report a case of successful emergency TIPS placement to control intra abdominal bleeding after blunt abdominal trauma in a patient with severe cirrhosis and portal hypertension. PMID- 10823546 TI - Fractured coronoid process and fractured olecranon with subluxation of the elbow in an adult. PMID- 10823547 TI - Acinetobacter calcoaceticus pneumonia and the formation of pneumatoceles. AB - Pneumatoceles are cystic lesions of the lungs often seen in children with staphylococcal pneumonia and positive-pressure ventilation. Acinetobacter calcoaceticus is an aerobic, short immobile gram-negative rod, or coccobacillus, which is an omnipresent saprophyte. The variant anitratus is the most clinically significant pathogen in this family, usually presenting as a lower respiratory tract infection. Acinetobacter has been demonstrated to be one of the most common organisms found in the ICU. We present three critically ill surgery patients with Acinetobacter pneumonia, high inspiratory pressures, and the subsequent development of pneumatoceles. One of these patients died from a ruptured pneumatocele, resulting in tension pneumothorax. Treatment of pneumatoceles should center on appropriate intravenous antimicrobial therapy. This should be culture directed but is most often accomplished with Imipenem. Percutaneous, computed tomographic-guided catheter placement or direct tube thoracostomy decompression of the pneumatocele may prevent subsequent rupture and potentially lethal tension pneumothorax. PMID- 10823548 TI - Blunt hemopericardium detected by surgeon-performed sonography. PMID- 10823549 TI - Cerebral air embolism caused by cardiopulmonary resuscitation after cardiopulmonary arrest on arrival. PMID- 10823550 TI - Juxtahepatic venous injuries: a critical review of reported management strategies. PMID- 10823551 TI - The image of trauma. Thoracic aortic transection secondary to blunt trauma. PMID- 10823552 TI - Bonegrafting from the proximal tibia. PMID- 10823553 TI - Critical analysis of injuries sustained in the TWA flight 800 midair disaster. PMID- 10823554 TI - Value of complete cervical helical computed tomographic scanning in identifying cervical spine injury in the unevaluable blunt trauma patient with multiple injuries: a prospective study. PMID- 10823555 TI - Dental unit water systems harbor large numbers of microorganisms. PMID- 10823556 TI - Report on a packaged handwashing antiseptic contaminated with Pseudomonas aeruginosa. PMID- 10823557 TI - Tuberculin skin testing in the era of multidrug-resistant tuberculosis. PMID- 10823558 TI - Are we doing enough to contain Acinetobacter infections? PMID- 10823559 TI - Epidemiology of nosocomial infections at Fukuoka University Hospital. PMID- 10823561 TI - Results of two observational studies in eight medical-surgical intensive care units in Germany to determine the frequency of hand washing by the medical staff and plot these results against the patient:personnel ratio PMID- 10823560 TI - Outbreak of Enterobacter cloacae related to understaffing, overcrowding, and poor hygiene practices. PMID- 10823562 TI - Surveillance of surgical-site infections: the world coming together? PMID- 10823563 TI - An operating surveillance system of surgical-site infections in The Netherlands: results of the PREZIES national surveillance network. Preventie van Ziekenhuisinfecties door Surveillance. AB - OBJECTIVES: To describe the results of the first year of the Dutch national surveillance of surgical-site infections (SSIs) and risk factors, which aims to implement a standardized surveillance system in a network of Dutch hospitals, to collect comparable data on SSIs to serve as a reference, and to provide a basic infrastructure for further intervention research. DESIGN: Prospective multicenter cohort study. SETTING: Acute-care hospitals in The Netherlands from June 1996 to May 1997. RESULTS: 38 hospitals participated, with a slight over-representation of larger hospitals. Following a total of 18,063 operations, 562 SSIs occurred, of which 198 were deep. Multivariate analysis of pooled procedures shows that age, preoperative length of stay, wound contamination class, anesthesia score, and duration of surgery were independent risk factors for SSI. When analyzed by procedure, the relative importance of these risk factors changed. Bacteriological documentation was available for 56% of the SSIs; 35% of all isolates were Staphylococcus aureus. Multiple regression analysis computed the mean extra postoperative length of stay associated with SSI to be 8.2 days. CONCLUSION: The first year of national surveillance has shown that it is feasible to collect comparable data on SSI, which are already used for education, policy, and decision making in the network of participating hospitals. This gives room to effectuate the next aim, namely to use the network as an infrastructure for intervention research. Multivariate analysis shows that feedback on a procedure specific level is important. PMID- 10823564 TI - Nasal carriage of Staphylococcus aureus is a major risk factor for surgical-site infections in orthopedic surgery. AB - OBJECTIVE: To determine the relative importance of different risk factors for the development of surgical-site infections (SSIs) in orthopedic surgery with prosthetic implants. DESIGN: In a cohort of 272 patients, the following possible risk factors were studied: age, gender, method of hair removal, duration of operation, surgeon, underlying illness, and nasal carriage of Staphylococcus aureus. Infections were recorded following the Centers for Disease Control criteria. The relation between risk factors and SSI was tested in univariate and multiple logistic regression analysis. SETTING: Community hospital in Breda, The Netherlands. RESULTS: 18 (6.6%) of 272 patients experienced SSI: 11 superficial and 7 deep SSI. These infections led in three cases to removal of the prosthesis and caused 286 extra days in hospital. The main causative pathogen was S aureus. In multiple logistic regression analysis, the following factors were independent risk factors for the development of SSI: high-level nasal carriage of S aureus (P=.04), male gender (P=.005), and surgeon 1 (P=.006). The only independent risk factor for SSI with S aureus was high-level nasal carriage of S aureus (P=.002). CONCLUSION: High-level nasal carriage of S aureus was the most important and only significant independent risk factor for developing SSI with S aureus. PMID- 10823565 TI - Identifying outliers of antibiotic usage in prevalence studies on nosocomial infections. AB - OBJECTIVE: To investigate whether the correlation between patients' antibiotic treatment (yes/no) and patients' infections (yes/no) in each hospital department, described by Pearson's correlation coefficient (p) for binary data as a measure for adequate use of antibiotics, is an appropriate quality indicator. DESIGN: Comparison of the results of repeated prevalence studies in different hospitals with the data of a national prevalence study, comparing the hospital (p) and reference (pNIDEP[Nosokomiale Infektionen in Deutschland: Erfassung und Pravention]) correlation coefficients for "use of antibiotics/ presence of infections." SETTING: The data of 5,377 surgical patients were separated from the total data of a national prevalence study in 72 representative hospitals to create a reference correlation coefficient (rhoNIDEP) with a reference range. Nine additional prevalence studies, involving a total of 4,984 patients, were repeatedly performed in the surgical departments of 8 other hospitals during a 12 month period, whereby the correlation coefficients rho(n) for every prevalence investigation were determined. RESULTS: In the national prevalence study, 15.3% of the surgical patients received antibiotics on the study day. Surgical patients had a 3.8% prevalence of nosocomial infections and a 7.0% prevalence of community acquired infections. Pearson's correlation coefficient rhoNIDEP for correlation between patients' binary data use of antibiotics and presence of infection was 0.62. To compare the correlation coefficient of each department with the appropriate reference range, the coefficients of the single departments were plotted against the number of patients; in these plots, three lines indicated the value rhoNIDEP and the upper and lower reference ranges, depending on the number of patients. Seven of eight surgical departments investigated during the repeated prevalence studies were found to be within the reference range, near the reference value, in the majority of prevalence studies; only one of the departments was identified as an outlier as regards antibiotic use. CONCLUSION: The correlation between patients' antibiotic treatment (yes/no) and patients' infections (yes/no) in hospitals or departments, as described by Pearson's correlation coefficient p for binary data with a definitive reference range depending on the number of patients, is useful for quality management in identifying the overall necessity for evaluating the indications for antibiotic use in one's own hospital. PMID- 10823566 TI - Nosocomial malaria from contamination of a multidose heparin container with blood. AB - A girl developed Plasmodium falciparum malaria in Riyadh, Saudi Arabia, a non malarious area. Twelve to 18 days before onset, she had been hospitalized for asthma on the same ward as three malaria patients. The only link between the malaria patients and the asthma patient was a multidose heparin container used to fill syringes for use on heparin locks and intravenous devices. Contamination of the heparin with blood occurred on at least one occasion when a needle had been left in place through the septum of this container and was used to refill a used syringe. PMID- 10823567 TI - Nosocomial bacteremia in HIV patients: the role of peripheral venous catheters. AB - A retrospective case-control study compared 40 human immunodeficiency virus (HV) infected patients with 43 nosocomial bacteremias (NB) to 77 HIV-infected patients without NB. Presence of a peripheral venous catheter (PVC) was associated with occurrence of NB and was significantly more frequent in NB without an identified source. PVCs probably are an underestimated source of NB in HIV-infected patients. PMID- 10823568 TI - Efficacy of a washer-pasteurizer for disinfection of respiratory-care equipment. AB - We evaluated the efficacy of a commercial washer-pasteurizer. Carriers were inoculated with 10(4) to 10(6) test organisms and pasteurized at 170 degrees F for 30 minutes. Pasteurization eliminated all test organisms with the exception of Bacillus subtilis spores. Pasteurization appears efficacious for the disinfection of respiratory-care equipment and could result in a cost savings of approximately $30,000 per year. PMID- 10823569 TI - Nosocomial Candida guilliermondii fungemia in cancer patients. AB - From 1988 to 1998, we identified nine patients with Candida guilliermondii fungemia. Four of the five patients with nosocomial infection died, while all of the non-nosocomial cases survived, even though one half of them (2/4) did not receive any treatment Nosocomial C guilliermondii fungemia is often associated with poor outcome despite aggressive antifungal therapy. PMID- 10823570 TI - Compliance with national recommendations for tuberculosis screening and immunization of healthcare workers in a children's hospital. AB - Physicians and clinical employees at a children's hospital were surveyed to compare their tuberculosis (TB) screening and immunization statuses. Failure to offer screening and immunization services to non-employee physicians was associated with lower rates of reported immunity to several vaccine-preventable diseases and with markedly lower rates of TB screening. PMID- 10823571 TI - Risk factors for nosocomial infections in a critically ill pediatric population: a 25-month prospective cohort study. AB - We studied risk factors for nosocomial infections among 500 critically ill children who were admitted to a pediatric intensive care unit from August 1994 through August 1996 and who were prospectively followed until death, transfer, or discharge. Age, gender, postoperative state, length of stay, device-utilization ratio, pediatric risk of mortality score, and total parenteral nutrition were the risk factors studied. Through multivariate analysis, we identified three independent risk factors for nosocomial infection: device-utilization ratio (odds ratio [OR], 1.6; 95% confidence interval [CI95], 1.10-2.34), total parenteral nutrition (OR, 2.5; CI95, 1.05 5.81) and length of stay (OR, 1.7; CI95, 1.31 2.21). PMID- 10823572 TI - Control of occupational hepatitis B among healthcare workers in the Czech Republic, 1982 to 1995. AB - Occupational hepatitis B remains a threat to healthcare workers (HCWs) worldwide, even with availability of an effective vaccine. Despite limited resources for public health, the Czech Republic instituted a mandatory vaccination program for HCWs in 1983. Annual incidence rates of acute hepatitis B were followed prospectively through 1995. Despite giving vaccine intradermally from 1983 to 1989 and intramuscularly as half dose from 1990 to 1995, rates of occupational hepatitis B decreased dramatically, from 177 cases per 100,000 workers in 1982 (before program initiated) to 17 cases per 100,000 in 1995. Among high-risk workers, the effect was even more dramatic (from 587 to 23 per 100,000). We conclude that strong public-health leadership led to control of occupational hepatitis B among HCWs in the Czech Republic, despite limited resources that precluded administering full-dose intramuscular vaccine for much of the program. Application of a similar program should be considered for other countries in regions that currently do not have a hepatitis B vaccination program. PMID- 10823573 TI - SHEA conference on antimicrobial resistance. Society for Healthcare Epidemiology of America. AB - Antimicrobial resistance is an increasing problem in healthcare institutions and in the community. Public concern about resistance is also increasing. The issue is broad and complex and not readily addressed by government, industry, or professional societies alone. On October 29-30, 1998, 19 representatives of various professional societies and governmental agencies met under the auspices of the Society for Healthcare Epidemiology of America (SHEA) at Brook Lodge Conference Center in Augusta, Michigan. The purpose of the meeting was to discuss the current status of antimicrobial resistance in the United States and Canada, including present society and governmental efforts to address the problem. Representatives exchanged experiences through presentations and discussions on the first day, then on the second day held a brainstorming session to address future needs and priorities in addressing the resistance problem. It was agreed that a national coordinated effort was needed. As part of this national effort, representatives called for the creation of a National Coalition on Antibiotic Resistance (NCAR) to combat antibiotic resistance through education, research, prevention, and advocacy. Priorities for NCAR were focused in four areas: (1) education of the public and professionals; (2) support of basic and applied research; (3) provision of an information resource and clearinghouse; and (4) advocacy initiatives. At the recommendation of the SHEA Board, discussions with the National Foundation for Infectious Diseases for the joint development of NCAR have begun. PMID- 10823574 TI - Ts65Dn mouse, a Down syndrome model, exhibits elevated myo-inositol in selected brain regions and peripheral tissues. AB - myo-Inositol is elevated in the Down syndrome (DS; trisomy 21) brain and may play a role in mental retardation. In the present study, we examined brain regions and peripheral tissues of Ts65Dn mouse, a recently characterized genetic model of DS, for abnormal myo-inositol accumulation. A GC/MS technique was used to quantitate myo-inositol and other polyol species (ribitol, arabitol, xylitol, and 1,5 anhydrosorbitol) in tissues from the Ts65Dn mice and control diploid mice. myo Inositol was found to be elevated in frontal cortex, hippocampus, and brain stem but not in cerebellum of the Ts65Dn mouse. Among peripheral organs examined, liver and skeletal muscle were found to excessively accumulate myo-inositol. In all tissues, concentrations of polyol internal controls were normal. The Ts65Dn mouse is useful to study the possible effect of elevated myo-inositol on cellular processes. PMID- 10823575 TI - Hyperammonemia impairs NMDA receptor-dependent long-term potentiation in the CA1 of rat hippocampus in vitro. AB - Hyperammonemia is considered the main factor responsible for the neurological and cognitive alterations found in hepatic encephalopathy and in patients with congenital deficiencies of the urea cycle enzymes. The underlying mechanisms remain unclear. Chronic moderate hyperammonemia reduces nitric oxide-induced activation of soluble guanylate cyclase and glutamate-induced formation of cGMP. NMDA receptor-associated transduction pathways, including activation of soluble guanylate cyclase, are involved in the induction of long-term potentiation (LTP), a phenomenon that is considered to be the molecular basis for some forms of memory and learning. Using an animal model we show that chronic hyperammonemia significantly reduces the degree of long-term potentiation induced in the CA1 of hippocampus slices (200% increase in control and 50% increase in slices of hyperammonemic animals). Also, addition of 1 mM ammonia impaired the maintenance of non-decremental LTP. The LTP impairment could be involved in the intellectual impairment present in chronic hepatocerebral disorders associated with hyperammonemia. PMID- 10823576 TI - Monoamine changes in the brain of BALB/c mice following sub-lethal infection with Nocardia asteroides (GUH-2). AB - BALB/c mice injected intravenously with a single, sub-lethal dose of Nocardia asteroides GUH-2 develop several levodopa responsive movement disorders. These included headshake, stooped posture, bradykinesia, and hesitation to forward movement. The changes in monoamine levels in the brain of these mice were determined. There was a significant loss of dopamine with greatly increased dopamine turnover in the neostriatum 7 to 29 days after infection. These effects were specific for dopaminergic neurons since minimal changes were found in neostriatal norepinephrine and serotonin even though serotonin turnover was increased. Changes in monoamine metabolism were not limited to the neostriatum. There were reduced levels of serotonin and norepinephrine with increased serotonin turnover in the cerebellum. One year after infection, dopamine metabolism had returned to near normal levels, but many of the movement disorders persisted. Specific changes in neurochemistry did not always appear to correspond with these impairments. Nevertheless, these data are similar to those reported in MPTP treated BALB/c mice. PMID- 10823577 TI - Conserved fatty acid composition of proteolipid protein during brain development and in myelin subfractions. AB - Myelin proteolipid protein (PLP) is modified after translation by the attachment of long-chain fatty acids to several cysteine residues. In this study, the amount and pattern of fatty acids covalently bound to rat PLP were determined during brain development and in myelin subfractions. For this purpose, PLP was isolated by gel-filtration chromatography in organic solvents, subjected to alkaline methanolysis, and the released fatty acid methyl esters were analyzed by gas liquid chromatography. At all ages examined, PLP had the same amount of covalently-bound fatty acids (3-4% w/w) and palmitate, oleate and stearate were always the major acyl chains. In contrast to myelin lipids, the fatty acid composition of PLP showed only minor changes between 15-days and 90-days of age. The amount and pattern of fatty acids bound to PLP prepared from three myelin subfractions were also indistinguishable. The conservation of a characteristic PLP-fatty acid make-up during brain development and in various myelin compartments suggests that this post-translational modification is essential for the normal functioning of the protein. PMID- 10823578 TI - The adenosine receptor agonist, APNEA, increases calcium influx into rat cortical synaptosomes through N-type channels associated with A2a receptors. AB - N6-2-(4-aminophenyl)ethyladenosine (APNEA) is a nonselective adenosine receptor agonist known to have a high affinity for the adenosine A1 and A3 receptors. It was found to be able to dose-dependently increase the sustained (4 min) Ca2+ influx into rat cortical synaptosomes while 2-chloro-N6-(3-iodobenzyl)-adenosine 5-N-methyluronamide (Cl-IB-MECA), a selective A3 agonist has no effect. However, this effect of APNEA was not affected by the presence of 8-cyclopentyl-1,3 dimethylxanthine (CPT), a selective A1 antagonist; but instead completely abolished by 8-(3-chlorostyryl)caffeine (CSC), a selective A2a antagonist, or omega-conotoxin GVIA. These results show that in the rat cortex, presynaptic A2a receptors can mediate neurotransmitter release by increasing Ca2+ influx through the N-type calcium channels. A1 and A3 receptors appear not to be involved. PMID- 10823579 TI - Changes of acetylcholinesterase activity in brain areas and liver of sucrose- and ethanol-fed rats. AB - The effects of chronic ethanol or sucrose administration to rats on acetylcholinesterase from brain and liver were investigated. Membrane-bound and soluble acetylcholinesterase activities were determined in fractions prepared by centrifugation. The thermal stability and the effects of temperature and different types of alcohols on acetylcholinesterase activity were also studied. Membrane-bound acetylcholinesterase activity increased (p < 0.01) in the liver after chronic ethanol administration, whereas no differences among groups in the encephalic areas, except in the brain stem soluble form, were found. Membrane bound acetylcholinesterase from the ethanol- and sucrose-treated groups was more stable at the different temperatures assayed between 10 and 50 degrees C than that corresponding to the control group. Non-linear Arrhenius plots were obtained with preparations of membrane-bound acetylcholinesterase from rat liver, with discontinuities at 30 degrees C (control or sucrose groups) or 34-35 degrees C (alcohol group). Assays made with membrane-bound or soluble enzyme from brain showed linear Arrhenius plots in all groups studied. The inhibitory effects of increasing concentrations of ethanol, n-propanol and n-butanol on acetylcholinesterase preparations from forebrain, cerebellum, brain stem and liver of the three experimental groups (control, sucrose-fed and ethanol-fed) were very similar. However, n-butanol displayed a biphasic action on particulate or soluble preparations of rat forebrain. n-butanol inhibited (competitive inhibition) at higher concentrations (250-500 mM), while at lower concentrations (10-25 mM), the alcohol inhibited at low substrate concentrations but activated at high substrate concentration. These results suggest that the liver is more affected by ethanol than the brain. Moreover, the lipid composition of membranes is probably modified by ethanol or sucrose ingestion and this would affect membrane fluidity and consequently the behaviour of acetylcholinesterase. PMID- 10823581 TI - Effect of isoprostanes on sympathetic neurotransmission in the human isolated iris-ciliary body. AB - Isoprostanes (IsoP's) are prostaglandin-like compounds that are derived from free radical catalyzed peroxidation of arachidonic acid independent of the cyclcooxygenase enzyme. In the present study, we investigated the effect of IsoP's on norepinephrine (NE) release from human isolated iris-ciliary bodies. Isolated human iris-ciliary bodies were prepared for studies of [3H]NE release using the superfusion method. Both 8-iso-prostaglandin F2alpha (F2-IsoP) and the thromboxane (Tx) receptor agonist, U46619 enhanced field-stimulated [3H]NE release from isolated, superfused human iris-ciliary bodies without affecting basal tritium efflux. On the other hand, an equimolar concentration (10 microM) of 8-iso-prostaglandin E2 (E2-IsoP) inhibited evoked [3H]NE overflow. The Tx receptor antagonist, SQ 29548 blocked the enhancements of electrically-evoked [3H]NE release induced by F2-IsoP and U46619. However, the inhibitory responses elicited by E2-IsoP was not antagonized by SQ 29548. We conclude that IsoP's can produce both excitatory and inhibitory effects on sympathetic neurotransmission in human isolated iris-ciliary bodies. The stimulatory effects of IsoP's on NE release may be mediated by Tx-receptors. PMID- 10823582 TI - The distribution of serotonergic nerves in microencephalic rats treated prenatally with methylazoxymethanol. AB - Prenatal exposure of pregnant rats to methylazoxymethanol acetate (MAM) induces microencephaly in the offspring. In the present study of these microencephalic rats (MAM rats) we used quantitative autoradiography to investigate [3H] paroxetine binding sites, which are a selective marker of serotonin (5-HT) transporters (5-HTT). The binding in the accumbens, cortex, hippocampus, and dorsolateral thalamus was significantly increased in MAM rats, compared to the control rats, while there was a significant decrease in the dorsal raphe nucleus of the MAM rats. The levels of 5-HTT mRNA in the dorsal raphe nuclei were analyzed by in situ hybridization, which revealed a significant decrease in 5-HTT mRNA-positive neurons in the MAM rats compared to the control rats. The results imply serotonergic hyperinnervation in the cerebral hemispheres of MAM rats, while a target-dependent secondary degeneration of 5-HT neurons might be induced in the dorsal raphe nuclei of MAM rats. PMID- 10823580 TI - Oxidation of polyamines and brain injury. AB - Several amine oxidases are involved in the metabolism of the natural polyamines putrescine, spermidine, and spermine, and play a role in the regulation of intracellular concentrations, and the elimination of these amines. Since the products of the amine oxidase-catalyzed reactions -- hydrogen peroxide and aminoaldehydes -- are cytotoxic, oxidative degradations of the polyamines have been considered as a cause of apoptotic cell death, among other things in brain injury. Since a generally accepted, unambiguous nomenclature for amine oxidases is missing, considerable confusion exists with regard to the polyamine oxidizing enzymes. Consequently the role of the different amine oxidases in physiological and pathological processes is frequently misunderstood. In the present overview the reactions, which are catalyzed by the different polyamine-oxidizing enzymes are summarized, and their potential role in brain damage is discussed. PMID- 10823583 TI - MPTP decreases MT-I mRNA in mouse striatum. AB - 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) is a drug that induces parkinsonism in humans and non-human primates. Free radicals are thought to be involved in its mechanism of action. Recently, metallothionein has been proposed to play a role as a scavenger of free radicals. In the present work, we studied the effect of MPTP neurotoxicity on brain metallothionein-I (MT-I) mRNA expression. Male C-57 black mice were treated with MPTP (30 mg/kg, i.p., daily) for 3 or 5 days. All animals were killed by cervical dislocation 7 days after the last MPTP dose. The brains were removed quickly and immediately frozen, and quantitative in situ hybridization was performed using MT-I cDNA probe. MT-I mRNA content in striatum, a region which is known to be highly predisposed and sensitive to MPTP-induced oxidative stress, decreased by 30% (3 days) and 39% (5 days) respectively, after the last MPTP administration. These results suggest that MT-I gene expression is decreased in MPTP neurotoxicity. It is suggested that the reduction of MT, an anti-oxidant and a free radical scavenger, in the striatum by MPTP enables the neurotoxin to exert maximal oxidative damage to the striatum. PMID- 10823584 TI - Measurement of apolipoprotein E (apoE) in cerebrospinal fluid. AB - Apolipoprotein E (apoE) is a protein involved in transport of lipids and has been implicated to play an important role in regeneration after nerve injury. Determination of apoE in cerebrospinal fluid (CSF) thus have a potential interest when studying different forms of brain damage and as a marker of ongoing regenerative processes in the brain. However, previous studies on CSF-ApoE in Alzheimer's disease (AD) have given inconclusive results. Such inconsistent results might be related to confounding factors interfering with sample handling and/or analyses, which have not been fully elucidated. We therefore examined different potential confounding factors for analyses of apoE in CSF and also developed a new enzyme linked immunosorbent assay (ELISA). The hydrophobic character of ApoE resulted in adsorption to different types of test tubes commonly used for collection of CSF at lumbar puncture, resulting in falsely low levels. This makes CSF handling critical, especially if samples are taken in different types of tubes, or is transferred to new tubes. Taking this confounding factors in consideration and analysing patient and control CSF handled in the same way and using the new ELISA, we could confirm our previous finding of reduced levels of ApoE in AD, (3.4 +/- 1.3 mg/l) compared with controls (4.5 +/- 2.7 mg/l) (p = 0.045). Both in the AD and in the control group, higher levels of CSF-ApoE was found in individuals possessing the ApoE4 alleles. Our results support that CSF-ApoE is reduced in AD, and that handling of CSF is a critical factor, which may explain the discrepant results from previous studies. Differences in the amount of patients and controls possessing the ApoE4 allele included might also increase the variance between different studies. PMID- 10823585 TI - Effect of inhibition of cyclooxygenase on pre- and postjunctional actions of peroxides in the iris-ciliary body. AB - In the present study, we investigated the effect of inhibition of cyclooxygenase (COX) with flurbiprofen (FBF) on peroxide-induced enhancement of field-stimulated [3H]norepinephrine ([3H]NE) release from bovine isolated irides. Furthermore, the effect of FBF was examined on peroxide-induced attenuation of contractions evoked by carbachol on this tissue. Irides were prepared for studies of neurotransmitter release and for measurement of contractile tension in vitro. Pretreatment of tissues with FBF (10 microM) caused significant (P < 0.001) rightward shifts of concentration-response curves to H2O2 and also decreased cumene hydroperoxide (cuOOH)-induced enhancement of evoked [3H]NE release. FBF (10 microM) partially prevented the attenuation of carbachol-induced contractions induced by H2O2 (300 microM) and cuOOH (300 microM). We conclude that inhibition of the biosynthesis of prostanoids reduced both the prejunctional stimulatory effects of H2O2 and cuOOH on sympathetic neurotransmission and inhibitory effects of peroxides on carbachol-induced contractions the in the bovine isolated iris. PMID- 10823586 TI - Contribution of MAP kinase pathways to the activation of ATF-2 in human neuroblastoma cells. AB - Activated Transcription Factor-2 (ATF-2) is important during development of and during injury to the brain. Both Jun N-terminal Kinases (JNKs) and p38 Mitogen Activated Protein Kinases (p38MAPKs) may phosphorylate ATF-2, but the contribution of these two pathways in cells has never been investigated. We have assayed endogenous p38MAPK activity in SK-N-MC and SH-SY5Y human neuroblastoma cells for activation of a GAL4/ATF-2 fusionprotein, by means of titrations of transfected expression plasmids and by using the p38MAPK inhibitor SB203580. It was found that basal activation of ATF-2 was independent of p38MAPK and that whereas MAPK kinase-3 (MKK3) was a weak inducer of ATF-2 activation, it was a potent activator of the stress activated transcription factor CHOP. In contrast, ATF-2 was very potently activated by the JNK pathway activator MAPK kinase kinase 1 (MEKK1). Thus, kinases downstream of MEKK1 appear relevant, but it is unlikely that p38MAPKs contribute quantitatively to activation of ATF-2 in these cells. PMID- 10823587 TI - Synthesis and release of dopamine in rat striatal slices: requirement for exogenous tyrosine in the medium. AB - When incubated in a tyrosine-free medium, the tissue dopamine (DA) level of rat striatal slices increased by about 921 +/- 15 pmol/mg protein during 90 min of preincubation. In contrast, the tissue-free tyrosine level declined only 130 pmol/mg protein in the same assay period. Depolarization of the slices with high K+ increased both DA and DOPAC outputs and depleted tissue DA level by about 75%. Although 60 min of resting after high K+ depolarization significantly restored the tissue DA levels, neither this restoration nor depolarization-induced DA release was altered by exogenous tyrosine. Similarly, failure of exogenous tyrosine was also observed during three successive depolarization periods of striatal slices. These results indicate that nigrostriatal dopaminergic neurons are able to synthesize and release the DA in the absence of exogenous tyrosine in the medium. Since the free tyrosine level in the slices does not seem to be a sufficient source, it is likely that tyrosine mobilized from its bound source(s) supports the DA synthesis under in vitro experimental conditions. PMID- 10823589 TI - Correction of limb deformities in the 21st century. PMID- 10823588 TI - Antiserum against S-100 protein prevents long term potentiation through a cAMP related mechanism. AB - Long term potentiation (LTP) was induced in the CA1 region of rat hippocampal slices by tetanization of the Schaffer collaterals. Local pretreatment of CA1 with serum of rabbits immunized against S-100 prevented the potentiation. However, treatment of the slices with a membrane permeant cAMP analogue, such as 8-Br-cAMP, could protect against the blocking effect of anti S-100 serum. We suggest that in the rat endogenous S-100b is involved in transduction mechanisms during LTP induction, via its ability to stimulate adenylate cyclase. Possible mechanisms of this action are discussed. PMID- 10823590 TI - Free fat interpositional graft in acute physeal injuries: the anticipatory Langenskiold procedure. AB - Free fat graft interposition has been used extensively in management of physeal injuries with established growth disturbances. The use of this technique as part of the management of acute physeal injuries has not been reported. Here we report on its application in acute physeal injuries, where it has prevented the formation of an anticipated physeal arrest. PMID- 10823591 TI - Peak height velocity as a maturity indicator for males with idiopathic scoliosis. AB - We retrospectively studied 43 adolescent boys treated with orthoses for idiopathic scoliosis to assess the usefulness of the timing of peak height velocity for predicting growth remaining and the likelihood of curve progression when compared with Risser sign, closure of the triradiate cartilage, and chronologic age. We compared the peak height velocity data in boys to our previous work for girls with adolescent idiopathic scoliosis. We found the median height velocity plots showed a similar high peak and sharp decline as is found in girls. All 13 patients with a curve magnitude > 30 degrees at the time of peak height velocity had progression of their scoliosis to > 45 degrees despite bracing. Four of 29 patients (14%) with curves < or = 30 degrees at peak height velocity progressed to 45 degrees. These values generate a sensitivity of 76%, specificity of 100% and accuracy of 91% in predicting progression to 45 degrees. Similar values have been found in female patients. The use of peak height velocity to predict the length of time for remaining growth was superior to Risser sign and chronologic age for boys with idiopathic scoliosis. Closure of the triradiate cartilage approximated the timing of peak height velocity in boys. PMID- 10823592 TI - Epiphyseal plate transplantation between sites of different growth potential. AB - The purpose was to study epiphyseal plate growth after microvascular transplantation to sites of different growth potential. The hypothesis was that the growth potential of an epiphyseal plate transplant is a function of the donor irrespective of the recipient site to which it is transplanted. Immature rabbits were used in an experiment that transplanted microsurgically revascularized second metatarsal epiphyseal plates. There were three experimental groups in which transplants were made to (i) sites of the same growth potential (orthotopic), (ii) sites of higher growth potential (proximal tibia), and (iii) sites of lower growth potential (third metacarpal). Control groups were nonoperated animals, animals with an anteromedial proximal tibial osteotomy, and nonoperated contralateral limbs of all experimental animals. Postoperative graft viability was checked with fluorochrome labeling. Postoperative growth was measured from serial standardized radiographs. Follow-up was for either 5 or 8 weeks. Animals were then killed, and epiphyseal plate specimens removed for histomorphometric analysis. Results showed that the total growth of experimental second metatarsal transplants was not statistically different (p > 0.05) in any recipient site. It was noted, however, that transplanted epiphyseal plates in all experimental groups grew at lower rates than nonoperated controls. PMID- 10823593 TI - The hip in adults with classic bladder exstrophy: a biomechanical analysis. AB - We studied the untreated pelvic deformity in 14 adult patients with bladder exstrophy to determine the relative hip-joint force and stress and their effects on the clinical status of the hip. Pelvic radiographs were used for biomechanical analysis to calculate joint force and joint stress (force/area) relative to partial body weight, which allowed comparison between patients and age-matched controls. IOWA hip ratings were used for clinical evaluation, and hips were scored radiographically for degenerative joint disease. The mean relative joint force and joint stress was significantly higher for exstrophy patients (p < 0.001). In addition, the mean distance from the body center to the center of the femoral head was significantly increased (p < 0.001) in bladder exstrophy patients. An increase in diastasis correlated with an approximate 30% increase in the distance from the center of the femoral head to the body midline. The mean distance from the greater trochanter to the femoral head center was significantly less (p < 0.02) and the center-edge angle was significantly decreased in exstrophy patients (p < 0.05). Two patients' hips showed significant subluxation, and one of these showed degenerative hip disease in association with poor IOWA and radiographic scores. The force and stress on the hip joint are increased in untreated adult bladder exstrophy patients. Further longitudinal study is indicated to validate these findings and to determine whether clinically important degenerative changes are occurring. This could affect treatment recommendations in childhood. PMID- 10823594 TI - Does early treatment by abduction splintage improve the development of dysplastic but stable neonatal hips? AB - A prospective trial was carried out to assess the outcome of children aged from 2 to 6 weeks with stable but dysplastic hips, treated with abduction splintage or by observation. Forty-four patients with 63 dysplastic hips were entered into the study and allocated into the two treatment groups at random. The ultrasound measured percentage acetabular cover in the splinted group improved in the first 3 months from an average of 32.8 to 54.3%. In the unsplinted group, the increase in cover was from 36.7 to 48.6%. The changes in cover for the splinted group were significantly more than those for the unsplinted group (p < 0.003) There was, however, no significant difference between the two groups in acetabular angle measurements on plain radiographs taken at 3 months. At 24 months, similarly, there was no significant difference in the acetabular angles of the two groups. These results support the view that stable dysplastic hips will correct with growth and that there is no sustained benefit from early splintage. PMID- 10823595 TI - Results of Pavlik harness treatment for neonatal hip dislocation as related to Graf's sonographic classification. AB - In this prospective study, 41 dislocated hips in 40 patients were classified according to Graf's sonographic classification. Of them, 29 hips were Graf type III and 12 hips Graf type IV. All were treated in a Pavlik harness to relocate the dislocated hip dynamically. In type III hips, this was successful in 97% and 50% in type IV hips. We conclude that Graf's classification in dislocated hips has prognostic significance in treatment with the Pavlik harness. PMID- 10823596 TI - The value of distal greater trochanteric transfer in the treatment of deformity of the proximal femur owing to avascular necrosis. AB - A series of 49 patients (55 hips) who had distal greater trochanteric transfer at the mean age of 13.9 years because of the deformity of the proximal femur after treatment for developmental dislocation of the hip was reviewed to verify the value of the procedure. Mean follow-up was 8.8 years. Clinical and radiologic assessment was supplemented with strain-gauging evaluation of the muscles involved. Good results were achieved in patients who had good range of movement or isolated restriction of abduction before the operation. After surgery, a 22% increase of abductors torque was found, the Trendelenburg sign disappeared in 30 individuals, and 15 patients regained normal gait. No complications were observed. Our study confirmed the value of distal transfer of the greater trochanter in treatment for proximal femur deformity owing to avascular necrosis after management of developmental dislocation of the hip and clarified indications for this procedure. PMID- 10823597 TI - Determination of hip position in the Pavlik harness. AB - Fourteen children treated for developmental dysplasia of the hip with a Pavlik harness were evaluated at the time of harness application with clinical examination, hip ultrasonography, and anteroposterior radiography. Ten orthopedic surgeons with experience levels varying from residents to experienced pediatric orthopedic surgeons evaluated the studies to determine hip position in the harness. Clinical examination agreed with hip ultrasonography for hip position in 100% of hips. Interpretation of radiographs agreed with ultrasonography in only 49% of cases in which the hip was judged to be dislocated and in 82% of cases in which the hip was judged to be reduced. When imaging is used to aid in assessing hip position during treatment with a Pavlik harness, ultrasonography appears to be superior to anteroposterior radiography for assessing hip position. PMID- 10823598 TI - Follow-up study of severe slipped capital femoral epiphysis treated with Dunn's osteotomy. AB - This study evaluated long-term clinical and radiologic results of Dunn's osteotomy carried out in 50 consecutive cases for severe slipped capital femoral epiphysis. The results of all the patients were analyzed with 4 years 6 months of follow-up. The clinical results were very good and good in 90% of the patients and were fair or poor in 10% of the patients. Seven patients had an important complication (total necrosis or chondrolysis), and the clinical evolution was studied. Dunn's procedure seems to be adequate to correct severe slips. PMID- 10823599 TI - Legg-Calve-Perthes disease and passive smoking. AB - We carried out a case-control study with 90 patients with Legg-Calve-Perthes disease (LCPD) and 183 normal children, as controls, selected at random to determine whether the condition of passive smoking is related to the disease. Seventy-one of 90 of the LCPD group (78.9%) were passive smokers. Only 79 of 183 (43.2%) in the control group were passive smokers (p = 0.00000). We did not find any statistical relationship between passive smoking and evolution of the condition (p = 0.42883), Catterall extension (p = 0.60544), final Stulberg result (p = 0.53201), or presence of sequelae (p = 0.53256). We also could not find any statistical difference between ages (p = 0.18). The odds ratio was 5.3203 (95% confidence interval 2.92-9.69). The association between LCPD and passive smoking, after controlling for age and gender, became significant (p = 0.0000). Thus the risk of LCPD in passive smoking children is more than five times higher than in children who are not exposed to smoke. It seems that passive smoking is a factor directly or indirectly associated with LCPD. PMID- 10823600 TI - Avascular necrosis in children with acute lymphoblastic leukemia. AB - This study describes the prevalence and demographics of avascular necrosis (AVN) in children with acute lymphoblastic leukemia (ALL). With improving survival of ALL patients on modern chemotherapy regimens, an increasing number of children with AVN will be presenting to orthopaedists. From 1991 to 1996, 202 patients were treated for ALL at a major tertiary pediatric cancer referral center. Eight patients (4.0%) subsequently developed AVN at an average of 30.0 months after beginning chemotherapy. A total of 27 documented joints were involved, with an average of 3.4 joints affected per patient diagnosed with AVN. The subset of patients with high-risk ALL who underwent an aggressive chemotherapy protocol was particularly susceptible to developing AVN. Six of 58 high-risk ALL patients (10.3%) developed AVN at an average of 18.5 months. As ALL patients now frequently survive into adulthood, orthopaedists will be increasingly called on to manage AVN affecting multiple joints in children and young adults. PMID- 10823601 TI - Which implant for proximal femoral osteotomy in children? A comparison of the AO (ASIF) 90 degree fixed-angle blade plate and the Richards intermediate hip screw. AB - We reviewed the clinical and radiologic results of 164 proximal femoral osteotomies in 132 children using the AO (ASIF) 90 degree fixed-angle blade plate and the Richards intermediate hip screw to establish the clinical and radiologic outcomes, establish the incidence of complications, and determine the relative indications and contraindications for the two implants. The incidence of complication in this series was 9% (15 complications in 14 osteotomies in 13 patients) including one bursitis, one wound breakdown with exposure of the underlying plate, five infections, two peroneal nerve palsies, one fractured shaft of femur, one fractured neck of femur, two technical errors, one painful nonunion, and one loss of fixation. There were five (3%) revisions in total. Fifty-six osteotomies in 44 patients were performed using the Richards intermediate hip screw, and 108 osteotomies were performed in 88 patients using the AO 90 degree fixed-angle blade plate. We found that both implants were effective with an acceptable rate of complications and revision surgery. PMID- 10823602 TI - Significance of the lateral epiphysis of the acetabulum to hip joint stability. AB - Normal development of the acetabulum is crucial to the growth and stability of the hip. Twenty-five New Zealand White rabbits (postweaning) weighing 1.5-2.2 kg were used. Both hips were exposed through an anterolateral approach. On the right hip, a fixed area of superolateral physeal cartilage was damaged with drills. Sham open reduction was performed for the left hip. Radiographic changes of the right hips were evident at 6 weeks. At 12 weeks, the right hip dislocated in a posterior and superior direction. The left hip remained normal. Histopathologic analysis correlated strongly with the radiographic findings. There were thinning of cartilage cells of the acetabulum, with disorganization. The acetabular roof was poorly formed. The lateral acetabular epiphysis is vital to the development of the acetabular roof. Damage to this epiphysis may result in acetabular maldevelopment and subsequent hip instability. We tested the hypothesis that abnormality in this epiphysis can give rise to abnormal acetabular development. PMID- 10823603 TI - External tibial torsion and the effectiveness of the solid ankle-foot orthoses. AB - The aims of this study were to determine the influence of external tibial torsion on the effectiveness of the ankle-foot orthoses (AFO) in children with lumbosacral myelomeningocele. Forty patients with normal tibial rotation and 18 patients with excessive external tibial torsion were evaluated with three dimensional gait analysis at their comfortable walking speed. The group with normal tibial rotation showed significantly greater knee extension and lower mean extension moment compared with the group with external tibial torsion (p < 0.05). The posteriorly and laterally deviated ground-reaction force relative to the knee flexion axis compromises the ability of this force to facilitate knee extension. Patients with torsional magnitudes >20 degrees demand close inspection as candidates for derotation osteotomy. The AFO will continue to stabilize the ankle foot complex, but improved knee motion, knee-extensor activity, and ultimately walking efficiency may be compromised. PMID- 10823604 TI - The influence of ankle-foot orthoses on gait and energy expenditure in spina bifida. AB - We examined the effect of ankle-foot orthoses (AFOs) on gait and energy expenditure in children with spina bifida. Nine boys and three girls, 6 to 16 years of age, took part in the study. There were four children each with L4, L5, and sacral level lesions. Each child underwent gait analysis and energy consumption studies with and without AFOs. Walking speed was faster with AFOs (mean, 58 m/min) than without (mean, 50 m/min, p < 0.01). Stride length improved significantly (p < 0.001) from 0.89 m barefoot to 1.08 m with AFOs. Double support time was decreased from 0.32 seconds barefoot to 0.28 seconds with AFOs (p < 0.05). The oxygen cost of walking was significantly better with (0.33 mL/kg/m) than without AFOs (0.41 mL/kg/m, p < 0.001). Hip flexion at initial contact was increased. Ankle kinematics were unchanged, but ankle power generation showed a significant improvement; from 0.5 W/kg barefoot to 1.3 W/kg with AFOs. Increased ankle power generation results in improved hip flexion and stride length, which contributes to increased walking speed and reduced oxygen cost. The stability conferred by the AFOs is reflected in the reduced time spent in double support. PMID- 10823605 TI - The effect of tethered cord release on scoliosis in myelomeningocele. AB - To understand better the effects of detethering on scoliosis in persons with myelomeningocele, 21 patients with spinal dysraphism and scoliosis who had undergone a detethering procedure were retrospectively reviewed. Fourteen girls and seven boys were followed for an average of 5 years (range, 2-10). Three patients had thoracic, 12 had lumbar, and six had sacral level myelomeningocele. Three patients (14%) had curves that improved, and six (29%) stabilized. Twelve patients (57%) progressed >10 degrees. Eighty-six percent of patients with initial curves of >40 degrees and 100% of thoracic level patients went on to require spinal fusion. Eight complications occurred in six patients. Although patients with curves <40 degrees may have some improvement after a detethering procedure, those presenting with curves >40 degrees or thoracic neurologic levels had no improvement in their scoliosis. PMID- 10823606 TI - Correlation of static to dynamic measures of lower extremity range of motion in cerebral palsy and control populations. AB - The question addressed in this study was whether static measurements of hip, knee, and ankle range of motion correlate to dynamic measurements of hip and knee function during gait. Range-of-motion measures of the lower extremities taken during physical examination (static variables) were recorded on 80 adolescents with cerebral palsy and 30 adolescent normal controls. Kinematic measurements collected during gait analysis (dynamic variables) were recorded on the same patients and controls. Results indicated no correlation greater than r = 0.50 (R2 < 0.25) between any static and dynamic variable for either group--cerebral palsy patients or controls. The lack of good correlation of these measures indicates static physical examination variables such as popliteal angle and straight-leg raise are not good predictors of dynamic gait, such as knee-extension and hip flexion variables measured during ambulation in controls or cerebral palsy populations. PMID- 10823607 TI - Adductor tenotomies in children with quadriplegic cerebral palsy: longer term follow-up. AB - Adductor tenotomy procedures for prophylaxis against severe hip subluxation in children with quadriplegic cerebral palsy have reported good to excellent success rates; however, the overall follow-up has been short. In a retrospective review designed to obtain longer term follow-up of adductor tenotomies in these children, we found that 58% of our patients had continued subluxation or dislocation with the eventual need for further surgery. These results differ significantly from those of previous studies. There was a trend among the patients who did well to have less preoperative subluxation than those who needed further intervention, but we did not find a statistically significant cut-off value that was predictive of success or failure. Age at the time of surgery also did not influence the outcome in our patient population. An 8-year average follow up of adductor tenotomies showed a higher loss of hip stability than previous mid range follow-up studies. PMID- 10823608 TI - Results of latissimus dorsi and teres major transfer to the rotator cuff in the treatment of Erb's palsy. AB - A retrospective study of the results of latissimus dorsi and teres major transfer in the treatment of Erb's palsy was conducted in 10 patients. All patients underwent release of the pectoralis major and transfer of the latissimus dorsi and teres major tendons to the rotator cuff at a mean age of 7 years and 2 months. Range of motion, patient satisfaction, and presence of any complications were evaluated postoperatively at a mean of 3 years. Active shoulder abduction improved from a mean of 72 degrees preoperatively to 136 degrees postoperatively. Postoperative shoulder active external rotation averaged 64 degrees. Subjectively, all patients participating in a questionnaire thought use of the operated-on extremity had improved since surgery. All but one patient were satisfied with the final outcome. Complications included one hypertrophic scar. Transfer of the latissimus dorsi and teres major to the rotator cuff in treatment of persistent Erb's palsy improves shoulder abduction and external rotation and results in a high degree of patient satisfaction. PMID- 10823609 TI - Abductor paralysis and external oblique transfer. AB - Eighteen patients with hip abductor paralysis owing to poliomyelitis underwent external oblique muscle transfer. Five male and 13 female patients with a mean age of 15 years 5 months (range, 10-23 years) with an average follow-up of 5 years (range, 2-11 years) were studied. An improvement in gait with decreased limp and swaying and increased ability to walk long distances was seen in 17 patients. Nine patients became free of their assistive ambulatory devices. Subjective improvement was observed in 16 patients (the improved appearance being the main element of satisfaction). According to a detailed scoring system, six excellent, six good, three fair, and three poor results were identified. There was very good correlation between the scoring system and the final outcome, making it a valuable predictive point system. The transfer was suitable for flail lower limb as well. PMID- 10823610 TI - Surgical management of spinal involvement in children and adolescents with Gaucher's disease. AB - Gaucher's disease is an uncommon hereditary glycolipid storage disorder characterized by the accumulation of glucocerebroside in the lysosomes of macrophages of the reticuloendothelial system. Skeletal manifestations are variable in severity and typically involve the long bones. Vertebral involvement is less well characterized, particularly in children and adolescents. We report on the surgical management of spinal involvement in four children and adolescents with Gaucher's disease; two for kyphotic deformity and two for kyphotic deformity associated with neurologic compromise. We recommend anterior spinal release with fusion and posterior spinal fusion with segmental instrumentation in cases of kyphotic deformity. In cases of spinal cord compromise at the apex of the kyphotic deformity with retropulsion of involved bone, anterior decompression also should be performed. Routine surveillance for spinal deformity in patients with Gaucher's disease is necessary to allow early intervention before the development of severe deformity and neurologic compromise. PMID- 10823611 TI - The short clavicle syndrome. AB - Congenital or developmental shorting of the clavicle is an uncommon and isolated condition leading to a tangential position of the scapula. It presents in childhood as abnormal posture and may be familial. It causes minimal disability and affected patients do not require treatment. PMID- 10823612 TI - Poland's syndrome in one identical twin. AB - Female twins were evaluated at the Shriners Hospital in Lexington, Kentucky. One twin was normal, and the other twin had the classic findings of Poland's syndrome, manifested by absence of the pectoralis major and symbrachydactyly of the right upper extremity. Buccal smears from each child were submitted for DNA testing. The test confirmed monozygosity with 99.9% probability. Some previous reports have stated that Poland's syndrome is an autosomal dominant, genetically determined trait, whereas others have maintained that there is no genetic association. The original case described by Poland in 1841 was his cadaver, and no family history was reported. This twin study provides strong evidence that the condition is not determined by gene transmission. PMID- 10823613 TI - A paradigm for the age-related treatment of knee dislocations in Larsen's syndrome. AB - Three cases of Larsen's syndrome in siblings are presented to illustrate a paradigm for the age-related treatment of deformity. Each child presented at a different age, with differing degrees of deformity and growth disturbance. Both growth and remodeling potential influenced the choice of treatment and outcome. The surgeon who undertakes the correction or prevention of these deformities must face the problem of and, it is hoped, use, growth potential from open physes and the progressive loss of remodeling capacity with advancing age. PMID- 10823614 TI - Severe osteopenia with recurrent fractures after bone marrow transplant for Wiskott-Aldrich syndrome: a case report. AB - Wiskott-Aldrich syndrome (WAS) is a rare inherited disorder characterised by thrombocytopenia, eczema, and immunodeficiency. Bone marrow transplantation (BMT) is a well-established modality of treatment now routinely used and often curative. We report the case of a boy who developed osteopenia and sustained multiple long-bone fractures over a 5-year period after bone marrow transplant for WAS. The femora and tibiae of both lower limbs were involved with a clinical presentation similar to osteogenesis imperfecta. After commencing calcitriol treatment at the age of 8 years, the patient has not sustained any further fractures. He is now 11 years old. Although short-term changes in bone metabolism after BMT have been documented, the occurrence of repeated fractures associated with osteopenia has not been previously reported. PMID- 10823615 TI - Outcomes of external fixation of pediatric femoral shaft fractures. AB - Thirty-seven femoral shaft fractures, in 33 patients, were treated with unilateral external fixation after reduction from 1992 through 1998. Ten girls and 23 boys ranged in age from 4 to 14 years. Thirteen children had multiple injuries, whereas 20 children had isolated fractures. Average follow-up was 3 years, 9 months, with only five children lost to follow-up. The average duration in fixator was 107 days. Thirty-six of 37 fractures healed, and there was one delayed union. There was minimal angulation, and limb-length inequality was generally <1 cm; 72.7% had pin-tract infections. Eight (21.6%) patients refractured; four occurred in the four patients with bilateral femur fractures. We agree with previous reports that external fixation remains a viable option for treatment of pediatric femoral shaft fractures. However, in our series, rate of refracture (21.6%) after removal of the external fixator is significantly higher than previously reported in literature. Children with bilateral femur fractures were at greatest risk. PMID- 10823616 TI - Features of femoral fractures in nonaccidental injury. AB - Femoral fracture is one of the most common long bone fractures owing to nonaccidental injury, but there is little evidence in the literature about their characteristics. It is important medicolegally to establish a diagnosis of child abuse. Fourteen femoral fractures from definite nonaccidental injury were analysed and compared with 33 femoral fractures caused by definite accident. The age, site, and fracture patterns were carefully studied. Of inflicted femoral fractures, 92.8% (13 of 14) occurred in children younger than 1 year old. On studying the fracture morphology, we conclude that there is no specific roentgenographic site or fracture pattern that allows differentiation between accidental and nonaccidental femoral fractures. PMID- 10823617 TI - Asymmetric hip deformity and subluxation in cerebral palsy: an analysis of surgical treatment. PMID- 10823618 TI - CT of parenchymal and bronchial tuberculosis. AB - Tuberculosis (TB) remains a common disease in the World. Its incidence has risen steadily since 1985, despite a preexisting continuous decreasing of its frequency due to an effective chemotherapy. Nonwhite people, socioeconomically disadvantaged, chronically debilitated groups and AIDS patients are the most concerned. Chest radiography remains the first imaging modality to evaluate TB. Widely radiographic appearances can be encountered, including normal chest X-ray. CT can be useful in all stages of the disease, particularly when clinical and radiological findings are in disagreement and/or when imaging findings are equivocal. CT should be proposed at the end of an effective antituberculous treatment to better subsequently detect fine lesions suggestive of reactivation TB. PMID- 10823619 TI - Spiral CT of pulmonary nodules: interobserver variation in assessment of lesion size. AB - The aim of this study was to assess interobserver variability in size determination of pulmonary nodules at spiral CT. Twenty-three patients with known pulmonary nodules (diameter 2-40 mm, mean diameter 7 mm) underwent spiral chest CT (collimation 5 mm, pitch 1). Images were reconstructed at 3- and 5-mm intervals (RI). Hard copies were analyzed by two radiologists who recorded every nodule with regard to location, diagnostic confidence ("definite," "probable") and nodule size in increments of 1 mm with specific attention to correct classification into one of three size classes (< or = 5 mm, 6-10 mm, > 10 mm). Interobserver variability was determined with Pearson's correlation coefficient and kappa measure. Of a total of 286 nodules, 103 nodules were found accordingly by both readers at 3 mm RI, and 96 at 5 mm RI. There was a good correlation of measurements (in millimeters) between both readers (Pearson's correlation coefficient: 0.89-0.95). Interobserver variability in categories was good at both reconstruction intervals (kappa: 0.61 at 3 mm, 0.74 at 5 mm RI) and very good (0.81) at 5 mm RI when uncertain nodules were excluded. Spiral CT allows reproducible size determination of pulmonary nodules as shown by good interobserver agreement in exact size measurement and categorization into three size classes. PMID- 10823620 TI - CT-guided core-needle biopsy in the diagnosis of mediastinal lymphoma. AB - The advent of radiologic guidance techniques for percutaneous biopsy has changed the approach to the routine diagnosis of mediastinal lymphoma. The aim of the present study was to evaluate the diagnostic accuracy of CT-guided percutaneous core-needle biopsy (PCNB) in the clinical management of patients with mediastinal lymphoma. The results of 49 CT-guided PCNB of mediastinal lymphoma performed under local anesthesia in 42 ambulatory patients were analyzed. A positive diagnosis of lymphoma was obtained in 30 of 42 patients, with an overall success rate of 71.5%. The technique was equally successful in the diagnosis of Hodgkin's and non-Hodgkin's lymphoma. There were no major complications. Percutaneous CT guided CNB of mediastinal lymphoma is a quick, safe, accurate, and efficient alternative to open biopsy in the evaluation of mediastinal lymphoma, mainly at presentation. It should become the preferred initial diagnostic procedure for obtaining histologic samples in patients with suspected mediastinal lymphoma. PMID- 10823621 TI - The CT fallen-lung sign. AB - On chest radiograph, the diagnosis of tracheobronchial tear is usually suspected because of the persistence of a pneumothorax after chest tube insertion. Since this radiographic pattern is nonspecific, the diagnosis is usually made by bronchoscopy and delayed. The fallen-lung sign consists in the fall of the collapsed lung away from the mediastinum occurring when the normal central bronchial anchoring attachment of the lung is disrupted. In contrast to the persistent pneumothorax, this sign is specific but rarely observed. Our purpose is to present the corresponding CT patterns observed in two cases of right stem bronchus tear, consisting in a caudal-dependent displacement of the right upper lobe bronchus which becomes obliquely oriented. PMID- 10823622 TI - Spontaneous reversibility of "pleural thickening" in a patient with semi-invasive pulmonary aspergillosis: radiographic and CT findings. AB - We present serial radiographic and CT findings of spontaneous reversibility of "pleural thickening" in a patient with proved semi-invasive pulmonary aspergillosis who developed bilateral intracavitary aspergillomas. To the best of our knowledge, this is the first report in the literature of this feature. Radiologists should be aware that pleural thickening in patients with semi invasive aspergillosis does not necessarily indicate irreversible pleural fibrosis. PMID- 10823623 TI - Contrast-enhanced magnetic resonance angiography in the evaluation of peripheral bypass grafts. AB - The aim of this study was to determine the potential of contrast-enhanced magnetic resonance (MR) angiography in the evaluation of peripheral bypass grafts. Digital subtraction angiography (DSA) served as a standard of reference. Thirty-five patients with previous bypass graft surgery underwent DSA and contrast-enhanced MR angiography within 2 weeks. MR angiography was performed using a three-dimensional fast gradient-echo sequence after administration of gadopentetate dimeglumine. Every leg was divided into 11 segments and scored in five categories of stenosis. MR angiography findings were compared with those of DSA. A total of 38 bypass grafts and 454 segments in 27 patients were included in the evaluation. In 33 (87%) bypass grafts stenosis grading with both methods corresponded, and in 5 (13%) cases stenosis was overestimated on MR angiography. Agreement in detection of hemodynamically significant stenosis (stenosis =50%) was 94.7% with a sensitivity of 100% and a specificity of 91.3%. In 340 (83.0%) vascular segments there was conformity in graduation, in 69 (16.88%), there was a difference of one or more grades on MR angiograms. Forty-four segments (9.6%) were not assessable due to technical limitations. Contrast-enhanced MR angiography is an useful noninvasive tool in the detection of failing peripheral vascular bypass grafts. PMID- 10823624 TI - Spiral CT in an acute spontaneous aorto-caval fistula. AB - A patient with a known infrarenal atherosclerotic aneurysm measuring 8 cm presented with sudden dyspnoe, lower back pain and enormous venous congestion of the lower extremities. Spiral CT rendered precise diagnosis and localisation of a large and acute spontaneous aorto-caval fistula and made early surgical intervention possible. Abdominal aorto-caval fistulas are rare complications and can occur after surgery, trauma or spontaneously in an atherosclerotic or inflammatory aneurysm. Spiral CT enables a rapid and precise diagnosis facilitating the planning of the therapeutic approach to this complication. PMID- 10823625 TI - Time-resolved two-dimensional thick-slice magnetic resonance digital subtraction angiography in assessing brain tumors. AB - The aim of this study was to evaluate clinical applicability of two-dimensional (2D) thick-slice, contrast-enhanced magnetic resonance digital subtraction angiography (MRDSA) with high temporal resolution in diagnosis of brain tumors. Forty-four patients with brain tumors including, 15 meningiomas, 8 gliomas, 6 metastatic tumors, 4 neuromas, and 2 hemangioblastomas, were studied with 2D MRDSA with frame rate approximately 1 s. Images were continuously obtained following the initiation of bolus injection of gadolinium chelates for 40 s and subtraction images were generated in a workstation. We evaluated visualization of normal cranial vessels on MRDSA and compared MRDSA and intra-arterial digital subtraction angiography (IADSA) with regard to hemodynamic information. Large cerebral arteries, all venous sinuses, and most tributaries were clearly visualized. A stain was present in hypervascular tumors including all 15 meningiomas and 2 hemangioblastomas on MRDSA. Presence of a stain demonstrated on MRDSA and that on IADSA coincided in 16 of 20 cases (Spearman rank correlation value was 0.85). The location, shape, and phase of the stain on MRDSA were similar to those on IADSA. Two-dimensional MRDSA with high temporal resolution has a unique ability to demonstrate cerebral hemodynamics, such as IADSA, and can play an important role in assessing brain tumors. PMID- 10823626 TI - Diagnostic value of the fast-FLAIR sequence in MR imaging of intracranial tumors. AB - The aim of this study was to quantify imaging characteristics of fast fluid attenuated inversion recovery (FLAIR) sequence in brain tumors compared with T1 postcontrast- and T2-sequences. Fast-FLAIR-, T2 fast spin echo (FSE)-, and T1 SE postcontrast images of 74 patients with intracranial neoplasms were analyzed. Four neuroradiologists rated signal intensity and inhomogeneity of the tumor, rendering of cystic parts, demarcation of the tumor vs brain, of the tumor vs edema and of brain vs edema, as well as the presence of motion and of other artifacts. Data analysis was performed for histologically proven astrocytomas, glioblastomas, and meningiomas, for tumors with poor contrast enhancement, and for all patients pooled. Only for tumors with poor contrast enhancement (n = 12) did fast FLAIR provide additional information about the lesion. In these cases, signal intensity, demarcation of the tumor vs brain, and differentiation of the tumor vs edema were best using fast FLAIR. In all cases, rendering of the tumor's inner structure was poor. For all other tumor types, fast FLAIR did not give clinically relevant information, the only exception being a better demarcation of the edema from brain tissue. Artifacts rarely interfered with evaluation of fast FLAIR images. Thus, fast FLAIR cannot replace T2-weighted series. It provides additional information only in tumors with poor contrast enhancement. It is helpful for defining the exact extent of the edema of any tumor but gives little information about their inner structure. PMID- 10823627 TI - Comparison of a conventional cardiac-triggered dual spin-echo and a fast STIR sequence in detection of spinal cord lesions in multiple sclerosis. AB - The current optimal imaging protocol in spinal cord MR imaging in patients with multiple sclerosis includes a long TR conventional spin-echo (CSE) sequence, requiring long acquisition times. Using short tau inversion recovery fast spin echo (fast STIR) sequences both acquisition time can be shortened and sensitivity in the detection of multiple sclerosis (MS) abnormalities can be increased. This study compares both sequences for the potential to detect both focal and diffuse spinal abnormalities. Spinal cords of 5 volunteers and 20 MS patients were studied at 1.0 T. Magnetic resonance imaging included cardiac-gated sagittal dual echo CSE and a cardiac-gated fast STIR sequence. Images were scored regarding number, size, and location of focal lesions, diffuse abnormalities and presence/hindrance of artifacts by two experienced radiologists. Examinations were scored as being definitely normal, indeterminate, or definitely abnormal. Interobserver agreement regarding focal lesions was higher for CSE (kappa = 0.67) than for fast STIR (kappa = 0.57) but did not differ significantly. Of all focal lesions scored in consensus, 47% were scored on both sequences, 31% were only detected by fast STIR, and 22% only by dual-echo CSE (n.s.). Interobserver agreement for diffuse abnormalities was lower with fast STIR (kappa = 0.48) than dual-echo CSE (kappa = 0.65; n. s.). After consensus, fast STIR showed in 10 patients diffuse abnormalities and dual-echo CSE in 3. After consensus, in 19 of 20 patients dual-echo CSE scans were considered as definitely abnormal compared with 17 for fast STIR. The fast STIR sequence is a useful adjunct to dual-echo CSE in detecting focal abnormalities and is helpful in detecting diffuse MS abnormalities in the spinal cord. Due to the frequent occurrence of artifacts and the lower observer concordance, fast STIR cannot be used alone. PMID- 10823628 TI - Symptomatic unilateral dolicho-ectasia of the intracranial arteries in a child. AB - Dolicho-ectasia of the left posterior cerebral artery and internal carotid artery in a 5-year-old boy is reported. There was compression of the brainstem, the mammillary bodies and the optic tract. The value of MR angiography in the diagnosis of this vascular anomaly is demonstrated. Dolicho-ectasia of more than one cerebral artery in a child is extremely rare. PMID- 10823629 TI - Value of (18F)-FDG positron emission tomography, computed tomography, and magnetic resonance imaging in diagnosing primary and recurrent ovarian carcinoma. AB - The aim of this study was to compare prospectively the accuracy of whole-body positron emission tomography (PET), CT and MRI in diagnosing primary and recurrent ovarian cancer. Nineteen patients (age range 23-76 years) were recruited with suspicious ovarian lesions at presentation (n = 8) or follow-up for recurrence (n = 11). All patients were scheduled for laparotomy and histological confirmation. Whole-body PET with FDG, contrast-enhanced spiral CT of the abdomen, including the pelvis, and MRI of the entire abdomen were performed. Each imaging study was evaluated separately. Imaging findings were correlated with histopathological diagnosis. The sensitivity, specificity and accuracy for lesion characterization in patients with suspicious ovarian lesions (n = 7) were, respectively: 100, 67 and 86% for PET; 100, 67 and 86% for CT; and 100, 100 and 100% for MRI. For the diagnosis of recurrent disease (n = 10), PET had a sensitivity of 100%, specificity of 50% and accuracy of 90%. The PET technique was the only technique which correctly identified a single transverse colon metastasis. Results for CT were 40, 50 and 43%, and for MRI 86, 100 and 89%, respectively. No statistically significant difference was seen. Neither FDG PET nor CT nor MRI can replace surgery in the detection of microscopic peritoneal disease. No statistically significant difference was observed for the investigated imaging modalities with regard to lesion characterization or detection of recurrent disease; thus, the methods are permissible alternatives. The PET technique, however, has the drawback of less accurate spatial assignment of small lesions compared with CT and MRI. PMID- 10823630 TI - Can magnetic resonance imaging predict the success of parturition in oxytocin induced pregnant women? AB - The aim of this study was to assess whether magnetic resonance imaging could predict the outcome of attempted vaginal delivery in a group of pregnant women whose parturition had to be induced by oxytocin. The signal intensity and morphology alterations in the cervix of 21 full-term pregnant women were analyzed before the induction of parturition. T2-weighted gradient echo sequences were utilized and signal intensity in the cervix was measured from the anterior and posterior lips of the cervix. An index indicating the brightness range of the cervix was formulated to overcome the effects of the individual intensity changes. Imaging features including the signal intensity and the evidence of effacement were correlated with the actual type of delivery performed. Images were also assessed visually by two independent radiologists. Statistical analysis of brightness indexes that were considered to have a predictive value as an indicator for possible delivery was not significant. However, visually assessed signal intensity of the cervix correlated strongly with the type of delivery. Effacement itself was the most reliable parameter in predicting the progress of the delivery. In conclusion, MR imaging seems to be useful for predicting normal parturition in full-term pregnant women who need oxytocin induction. However, the presence of effacement seems to be a more reliable and practical parameter that will be preferred in that prediction. PMID- 10823631 TI - Cirsoid renal arteriovenous malformation treated by endovascular embolization with n-butyl 2-cyanoacrylate. AB - The authors report a rare case of renal arteriovenous malformation (rAVM) which was diagnosed by arteriography years after onset of intermittent haematuria. The rAVM of the cirsoid type was superselectively catheterized and embolized in toto with n-butyl 2-cyanoacrylate. Diagnostic imaging modalities and the technique of embolization are discussed. PMID- 10823632 TI - Endometrial stromal sarcoma of the uterus: MR and US findings. AB - We describe the MRI and US features of two patients with endometrial stromal sarcoma of the uterus. Both lesions appeared as voluminous polypoid masses within an expanded endometrial cavity on both US and MRI. They had mixed echo-texture and heterogenous signal intensity on both T1- and T2-weighted sequences. T2 weighted images were most helpful in detecting the endometrial nature of the disease and its relationships with surrounding myometrium. PMID- 10823633 TI - Uterine endometrial stromal sarcoma located in uterine myometrium: MRI appearance. AB - Two cases of uterine endometrial stromal sarcoma whose main mass was located in uterine myometrium are reported. They mimicked uterine leiomyoma with cystic degeneration or uterine leiomyosarcoma. Endometrial stromal sarcoma should be suggested in the differential diagnosis of mass lesion in uterine myometrium. PMID- 10823634 TI - Spontaneous renal artery dissection revealed by helical CT angiography. AB - Spontaneous renal artery dissection is rare and may be misdiagnosed because its clinical presentation is confusing. Diagnosis is usually made by intra-arterial angiography. We report a case where a spontaneous renal artery dissection was initially misdiagnosed as a renal abscess. Diagnosis was made later with helical CT for suspicion of renal artery stenosis in a patient with recent onset of hypertension. This unusual case shows that helical CT can be useful for non invasive diagnosis of renal artery dissection. PMID- 10823635 TI - CT colonography and MR colonography: current status, research directions and comparison. AB - With the introduction of multidetector technology in CT and the moveable table with integrated coil modules in MRI, the concept of multiorgan screening has become realistic. CT colonography and MR colonography are new radiologic techniques that promise to be highly sensitive colorectal screening examinations. This article reviews the current status and research directions in CT colonography and MR colonography, and compares these methods. PMID- 10823636 TI - Embolization for gastrointestinal hemorrhages. AB - Retrospective evaluation of interventional embolization therapy in the treatment of gastrointestinal hemorrhage over a long-term observation period from 1989 to 1997. Included in the study were 35 patients (age range 18-89 years) with gastrointestinal bleeding (GI) referred for radiological intervention either primarily or following unsuccessful endoscopy or surgery. Sources of GI bleeding included gastric and duodenal ulcers (n = 7), diverticula (n = 3), erosion of the intestinal wall secondary to malignancy (n = 6), vascular malformations (n = 4), and hemorrhoids (n = 2), as well as from postoperative (n = 6), posttraumatic (n = 2), postinflammatory (n = 4) or unknown (n = 1) causes. Ethibloc (12 cases) or metal coils (14 cases) were predominantly used as embolisates. In addition, combinations of tissue adhesive and gelfoam particles and of coils and Ethibloc were used (six cases). Finally, polyvinyl alcohol particles, a coated stent, and an arterial wire dissection were utilized in one case each. Bleeding was stopped completely in 29 of 35 cases (83%). In one case (3%) the source of bleeding was recognized but the corresponding vessel could not be catheterized. In five other cases (14%) there was partial success with reduced, though still persistent, bleeding. The rate of complications was 14%, including four instances of intestinal ischemia with fatal outcome in the first years, and, later, one partial infarction of the spleen without serious consequences. Gastrointestinal hemorrhage can be controlled in a high percentage of patients, including the seriously ill and those who had previously undergone surgery, with the use of minimally invasive interventional techniques. The availability of minicoils instead of fluid embolization agents has reduced the risk of serious complications. PMID- 10823637 TI - Tissue transition projection (TTP) of the intestines. AB - Tissue transition projection (TTP) represents a three-dimensional reconstruction technique for volumetric image data sets. To demonstrate the principle characteristics of TTP, a simple phantom consisting of two pipes with a simulated, wall-adherent polyp was scanned with spiral CT, and images were reconstructed by means of volume rendering for both opaque surface reconstructions and TTP. Tissue transition projection was used in 7 patients for reconstruction of the small intestine or the colon. Unlike three-dimensional reconstructions with opaque surfaces, TTP enhances surface transitions while suppressing homogeneous areas, allowing delineation of the bowel wall similar to conventional double-contrast studies. PMID- 10823638 TI - Long-standing painless intussusception in adults. AB - Long-standing painless intussusception in adults is considered to be rare. We report three such cases with an emphasis on color Doppler results. In our three cases the indication for abdominal US was a palpable mass in all these cases, and intussusception was detected by US at a time when the patients had only very mild abdominal discomfort. Persistence of sufficient blood flow, as was suggested by the color Doppler results, was thought to be the most likely pathomechanism of long-standing painless intussusception. The underlying disease was tuberculosis in two of the three cases. Thus, when encountering patients with painless intussusception, tuberculosis must be kept in mind. PMID- 10823639 TI - Malignant peritoneal mesothelioma presenting with respiratory symptoms. AB - Malignant peritoneal mesothelioma is a rare disease associated with mild, nonspecific abdominal symptoms and a wide spectrum of imaging findings, with thickened mesentery and peritoneum being the most common ones. A case of a malignant peritoneal mesothelioma presenting with manifestations of pulmonary disease is reported. Imaging evaluation revealed pleural, lung and pericardial involvement together with retroperitoneal lymphadenopathy, little ascites and extensive omental, but only subtle, mesenteric thickening. PMID- 10823640 TI - Intra-abdominal desmoplastic small round-cell tumor: a rare cause of peritoneal malignancy in young people. AB - We report the CT and pathological findings of an intra-abdominal desmoplastic small round-cell tumor in a young man. Computed tomography showed an extensive peritoneal and mesenteric disease with gross bulky masses, direct liver invasion, and lymph node involvement. This entity should be considered in the differential diagnosis of a young male patient presenting with features of widespread peritoneal malignant disease. PMID- 10823641 TI - Congenital absence of portal vein with nodular regenerative hyperplasia of the liver. AB - Congenital absence of the portal vein is a very rare anomaly. The intestinal and splenic venous drainage bypasses the liver and drain into the inferior vena cava (IVC). Two cases of such anomaly are described. Both cases were investigated by US coupled with echo-colour Doppler examination, CT and MR imaging, followed by digital subtraction angiography (DSA) and liver biopsy. In the first case the splenic and superior mesenteric vein formed a venous trunk which emptied directly into the IVC; in the second case, the splanchnic blood flowed into a dilated hepatofugal inferior mesenteric vein which connected to the left internal iliac vein. In both cases nodular regenerative hyperplasia of the liver was present, presumably due to an abnormal hepatic cell response to the absent portal flow. The particular contribution of MR imaging to the diagnosis of both vascular abnormalities and liver parenchyma derangement and its advantages over the other diagnostic techniques is emphasized. The clinical and radiological features of 17 previously reported cases are reviewed. PMID- 10823642 TI - Quantitative ultrasound at the hand phalanges: comparison with quantitative computed tomography of the lumbar spine in postmenopausal women. AB - The aim of our study was to evaluate the diagnostic effectiveness of a quantitative ultrasound technique for measuring bone tissue at the proximal phalanges of the non-dominant hand. We correlated the mean value of the amplitude dependent speed of sound (AD-SoS) and the ultrasound bone profile score (UBPS) measured at the phalanges with bone mineral density (BMD) of the lumbar spine gauged with quantitative computed tomography (QCT). We studied a group of 177 postmenopausal women consecutively presenting for osteoporosis screening. We observed that in the whole study group both AD-SoS and UBPS correlated weakly with BMD, although with statistical significance (r = 0.54 and r = 0.45, respectively; p < 0.0001). The same parameters showed a poor correlation (r = 0.48, p < 0.0001; and r = 0.23, p = 0.017) in the group of patients which QCT has classified as osteoporotic. Finally, no correlation was found in the rest of our population. The correlation coefficients of comparison between our QUS and QCT measurements are not strong enough to allow us a prediction of one measure from the other. Therefore, we do not consider the phalangeal osteosonography a valid substitute of QCT technique for assessment of bone status in postmenopausal women. PMID- 10823643 TI - Osteochondroma: MR imaging of tumor-related complications. AB - Osteochondromas can be complicated by mechanical irritation, compression or injury of adjacent structures, fracture, malignant transformation, and postoperative recurrence. Magnetic resonance imaging represents the most valuable imaging modality in symptomatic cases, because it can demonstrate typical features of associated soft tissue pathology, which can be differentiated from malignant transformation. Reactive bursae formation presents as an overlying fluid collection with peripheral contrast enhancement. Dislocation, deformation, and signal alterations of adjacent soft tissue structures can be observed in different impingement syndromes caused by osteochondromas. Magnetic resonance imaging provides excellent demonstration of arterial and venous compromise and represents the method of choice in cases with compression of spinal cord, nerve roots, or peripheral nerves, depicting changes in size, position, and signal intensity of the affected neural structures. Malignant transformation as the most worrisome complication occurs in approximately 1% of solitary and 5-25% of multiple osteochondromas. Magnetic resonance imaging is the most accurate method in measuring cartilage cap thickness, which represents an important criterion for differentiation of osteochondromas and exostotic (low-grade) chondrosarcomas. Cartilage cap thickness exceeding 2 cm in adults and 3 cm in children should raise the suspicion for malignant transformation. Finally, MR imaging can detect postoperative recurrence by depiction of a recurrent mass presenting typical morphological features of a cartilage-forming lesion. PMID- 10823644 TI - Recurrence of non-Hodgkin's lymphoma isolated to the right masticator and left psoas muscles. AB - We present the clinical and magnetic resonance imaging findings of a patient who, following treatment for pancreatic non-Hodgkin's lymphoma (NHL), relapsed with apparently isolated involvement of the right masticator space and left psoas muscles. Non-Hodgkin's lymphoma arising from the masticator space muscles is very rare. In addition, simultaneous lymphomatous involvement of multiple discrete skeletal muscle sites, in the absence of disease elsewhere, has previously only been reported in the limb or limb girdle muscles. Lymphoma should be considered as a cause of isolated enlarged skeletal muscles, even when involving such distant sites. PMID- 10823645 TI - Variations of sphenoid and related structures. AB - The aim of this study was to delineate the precise relationship between the sphenoid sinus and internal carotid artery and the optic nerve, as well as to assess incidence of the anatomic variations of these structures. A review of 92 paranasal sinus tomographic scans was made for anatomic variations of the sphenoid sinus and related bony and neurovascular structures. Coronal and axial tomographic sections were obtained with 2.5-mm section thickness. We assessed the protrusion of the internal carotid artery (ICA) and the optic nerve (ON) into the sphenoid sinus, bone dehiscence of these structures, and pneumatization of the anterior clinoid process (ACP) and pterygoid recess (PR), as well as the variations of the sphenoid sinus septum. The protrusion of the ICA into the sphenoid sinus was found in 24 (26.1%) patients. An ON protrusion was present in 29 (31.5%) patients. Pneumatization of the PR was encountered in 27 (29.3%) patients. There was not a statistically significant relationship between the pneumatization of the PR and ICA protrusion into the sphenoid sinus (chi2 = 0.258, p = 0.168). A significant relationship between the ACP pneumatization and protrusion of the ON into the sphenoid sinus was found (chi2= 0.481,p = 0.007). Preoperative recognition of the anatomic variations by the radiologist is beneficial for identification of the limits of dissection. This is particularly important in the sphenoid sinus area where extensive pneumatization of the skull base bones may distort the anatomic configuration. Therefore, axial and coronal CT sections should always be obtained prior to any surgery in the sphenoid sinus area. PMID- 10823646 TI - Antrochoanal polyps in children: CT findings and differential diagnosis. AB - Antrochoanal polyp (Killian polyp) is an infrequent, usually solitary, benign, slowly growing lesion that arises from the maxillary antrum and reaches the choana. These polyps have a discrete male predominance and are diagnosed usually between the third and the fifth decades of life. This report is based on three cases of antrochoanal polyp, occurring in the pediatric group, and the objective is to demonstrate their different CT characteristics, principal differential diagnoses, and potential complications. We emphasize that in all three cases of our series the growth of the polyp to the choana is through the accessory ostium. PMID- 10823647 TI - A novel "cooled-wet" electrode for radiofrequency ablation. AB - In the light of growing demands for improved applicability of radiofrequency ablation (RFA), recently we have developed a novel "cooled-wet" electrode by taking the advantages of both internally cooled and saline-enhanced electrodes. The efficacy of the electrode was evaluated in both ex vivo and in vivo liver RFA under both low and high power output levels. The ablation volume created with the "cooled-wet" electrode appeared to be much larger than that reported up to now with the use of other monopolar electrodes. The mechanisms on how this device optimizes the RF energy delivery are also discussed. PMID- 10823648 TI - Chronic osteomyelitis of the femur: value of PET imaging. AB - The purpose of this report is to discuss FDG-PET as a potentially new imaging tool in the diagnosis of infections of osteosynthetic material. We present a patient with a poly-trauma who developed a chronic osteomyelitis and ostitis after repeated osteosynthesis in a fibular transplant to the left femur. Work up included MRI, antigranulocyte antibody scintigraphy and positron emission tomography (PET) with F-18 fluorodeoxyglucose (FDG). Infection of the fibular transplant was demonstrated clearly by PET but not by the other methods. Positron emission tomography may become an important indication in the diagnosis and follow-up of bone infection. PMID- 10823649 TI - Radiology on Internet: advice in consulting websites and evaluating their quality. AB - Services offered by Internet are increasing continually and 15,000 medical websites covering all the specialties are available presently. Finding relevant information with a spider-web organization is difficult. We recommend starting with hierarchical lists which propose a selection of sites corresponding to medical specialties. We give our selection of important radiological websites dealing with image databases, case reports, radioanatomy and continuing medical education. While surfing the Web, an evaluation of the quality of websites is necessary. We used quality criteria proposed by Darmoni to rate the quality of ten websites. The global quality is good; help pages and external links are the main noticed shortcomings. All webmasters should pay attention to quality criteria and show visible marks of Darmoni criteria on their home page. To be able to foresee which Internet options should be developed, we evaluated our website and sent a questionnaire to our users. Clinical cases are the most requested facilities; they could be used for continuing medical education. PMID- 10823650 TI - The changed preliminary report: a repeatedly missed paediatric tibial tumour. AB - This report describes a malpractice case involving a delayed diagnosis of a malignant bone tumour in the proximal tibia in a 10-year-old child. This was caused by a combination of factors. The final report on the first examination failed to reach the patient files, and two subsequent X-ray exams failed to diagnose the tumour, due to misinterpretation in one and obscuring plaster of Paris in the other. PMID- 10823651 TI - Subcutaneous extension of a large psoas muscle Echinococcus cyst. PMID- 10823652 TI - MRI of intravascular leiomyomatosis extending to the heart. PMID- 10823653 TI - Anomalous pulmonary vein connection. PMID- 10823654 TI - Hydatid bronchial impaction: CT findings. PMID- 10823655 TI - Analysis of neutrophil proteins of patients with Behcet's disease by two dimensional gel electrophoresis. AB - Protein changes in the neutrophils of patients with Behcet's disease were analyzed by high resolution two-dimensional gel electrophoresis to investigate the pathological features of Behcet's disease. Two clear protein spots were found to be differently expressed between healthy volunteers and patients with Behcet's disease. One was a 53 kDa protein with pI 5.2 that was detected in healthy volunteers but was nearly absent in the patients. The other was a 40 kDa protein with pI 5.2 that was detected in the patients but nearly absent in the healthy volunteers. Analysis of the N-terminal amino acid sequence of the 40 kDa protein revealed that it was a truncated actin with an N-terminus of Met-44. The presence of the truncated actin in the neutrophils of patients was confirmed by Western blot analysis using an antibody to the C-terminus of actin. The 53 kDa protein could not be identified because its N-terminus was blocked. The presence of the truncated actin in the neutrophils of the patients may be important in understanding the pathology of Behcet's disease. PMID- 10823656 TI - Preparation of monoclonal antibody against ginsenoside Rf and its enzyme immunoassay. AB - A rapid and sensitive indirect competitive enzyme immunoassay method has been developed for quantitating ginsenoside Rf (Rf) in crude total Panax ginseng saponins and in rat plasma using high titer mouse monoclonal antibody (mAb) raised against a conjugate of Rf and bovine serum albumin (BSA). The isotype of mAb against Rf was IgG3 with a K chain. The presence of Rf inhibited the binding of the mouse anti-Rf mAb to a Rf-BSA solid phase coating antigen. The working range was 0.01-10 ng/assay and detection limits were 20 pg in various ginseng extract fractions or 34 pg in rat plasma per assay. The anti-Rf mAb cross-reacted with ginsenoside Rg2 by 57.5%, but not with other ginsenosides. However, this anti-Rf mAb did not cross-react with BSA or cellubiose, which is a carbohydrate component of Rf. Using this standard curve, we could measure the amount of Rf in ginseng total extract, ginseng total saponins, protopanaxadiol saponins, and propanaxatriol saponins. We could also measure the amount of Rf in rat plasma after the oral administration of Rf and found that Rf reached a maximum level in rat plasma after 16 h. These results indicate that the anti-Rf mAb could be useful for the quantitation of Rf in crude ginseng fractions and in body fluids. PMID- 10823657 TI - Cloning of the gene of beta-N-acetylglucosaminidase from Lactobacillus casei ATCC 27092 and characterization of the enzyme expressed in Escherichia coli. AB - The gene encoding beta-N-acetylglucosaminidase (GlcNAcase) of Lactobacillus casei ATCC 27092 was cloned and expressed in Escherichia coli. The gene consisted of 1581 nucleotides, and had a promoter, Shine-Dalgarno, and rho-independent type transcription termination sequences typical in bacteria. The protein deduced from the sequence consisted of 526 amino acids, and had a putative signal peptide of 14 amino acids and 5 possible asparagine-linked glycosylation sites. A conserved sequence was homologous to the 12 other hexosaminidases from different origins. The recombinant GlcNAcase (r-GlcNAcase) purified from the transformed E. coli had a MW of 39 kDa and lacked oligosaccharide chains. The isoelectric point and the optimum pH for the activity of r-GlcNAcase were similar to those of original GlcNAcases (o-GlcNAcase). However, the thermal stability was lower, and sensitivity to Cd2+, Fe2+, Cu2+ and sodium dodecyl sulfate (SDS) was higher than those of o-GlcNAcases, suggesting that the oligosaccharide moieties of the enzyme contribute to their stability. The Km value for p-nitrophenyl-N-acetyl-beta-1,4-D glucosamine (PNP-GlcNAc) of r-GlcNAcase (6.4 microM) implied that the affinity of r-GlcNAcase for the substrate was 200-fold higher than that of the original ones. PMID- 10823658 TI - Appearance of extracellular glutathione peroxidase (eGPx) in the ascite fluid of casein-elicited rats. AB - Glutathione peroxidase (GPx) activity was detected in the ascite fluid of rats injected intraperitoneally with 2.5% heat-denatured casein solution. Activity in the ascite fluid increased with time after the injection of casein, and reached a maximum at 24 h. The active component was concentrated with successive 35% ammonium sulfate precipitation and Activated Thiol-Sepharose 4B column chromatography from the ascite fluid of rats at 24 h after the injection of casein. No N-terminal amino acid of the protein corresponding to GPx was detected by automatic amino acid sequence analysis following separation with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transfer to a polyvinyl difluoride (PVDF) membrane. Following BrCN treatment of the protein, the N-terminal amino acid sequences of two 14 and 2.6 kDa peptide fragments were found to be S-G-T-I-Y-E-Y-G-A-L and K-I-H-D-I-R-W-N-F-E, respectively. The former and the latter fragments corresponded to sequences beginning at the 37th and 176th amino acid residues of rat extracellular GPx (eGPx), respectively. The exclusive presence of eGPx in the ascite fluid of rats elicited by casein was confirmed immunologically by ELISA, immuno-precipitation and Western blotting assays. No other GPx isozymes such as cytosolic GPx (cGPx), phospholipid hydroperoxide GPx (PHGPx) or intestinal GPx (iGPx) were detected. eGPx activity and protein were also detected in the pleuritic fluid of rats following injection of 2% carrageenan. These findings indicate that eGPx appears at various sites of acute inflammation in rats. This pattern is due to leakage from circulation as a result of the increased capillary permeability at inflammation sites elicited by chemotactic factors. PMID- 10823659 TI - Properties of NADPH oxidase in specific granule-rich fraction prepared from guinea pig neutrophils. AB - Both the plasma membrane-rich fraction and specific granule-rich fraction prepared from human neutrophil lysate by Percoll centrifugation have been reported to contain cytochrome b558, a membrane activation factor for NADPH oxidase. In this study, the plasma membrane-rich fraction and specific granule rich fraction of guinea pig neutrophils were prepared, and the abilities of both fractions to activate NADPH oxidase in a cell-free system consisting of either fraction, cytosol and arachidonate were compared. There was no difference in the Km value for NADPH between NADPH oxidase activated by specific granules or by plasma membranes. Optimum concentrations of arachidonate for the activation of NADPH oxidase in both the fractions were also the same. However, after freeze thawing, the specific granules markedly lost the ability, compared to plasma membranes. Such instability of specific granules was also observed on hypotonic- or deoxycholate-treatment. The inactivation by freeze-thawing was not suppressed by proteinase inhibitors, and gp91-phox, a large subunit of cytochrome b558, was not degraded by freeze-thawing. Freeze-thawed specific granules did not affect the ability in plasma membranes, indicating the absence of an inactivating factor in specific granules. The increase in the amount of cytosol in the cell-free assay mixture did not compensate for the markedly decreased ability of freeze thawed specific granules. Translocation of p47-phox, one of the cytosolic activation factors, to specific granules was not affected by freeze-thawing. We found that the ability of specific granules to activate NADPH oxidase was fragile, though it is unclear what is responsible for the instability, at present. PMID- 10823660 TI - Purification of a sensory-specific protein of 35 kDa (SSP-35). AB - Protein purification is a key technique for the identification of novel sensory nerve-specific proteins. A 35 kDa protein, the sensory neuronal specific protein, was isolated and purified from rabbit spinal ganglia and sensory fibers by homogenizing and deriving soluble extracts, followed by ion exchange chromatography using DEAE-Sephacel and gel filtration HPLC. Western blot analysis showed that the protein was present in spinal sensory ganglia but not in spinal motor neurons with anti-rat 35 kDa protein polyclonal antibody. The sensory specific protein 35 kDa is termed SSP-35 in this paper. We found that the purified SSP-35 promoted axonal growth of the dorsal root ganglia of chick E8 embryos. Our data reveal that the protocol is an effective method for the purification of SSP-35. The protein may not only be a useful marker for sensory neurons, but also a possible tool to study the regeneration and function of sensory neurons. PMID- 10823661 TI - Increased degradation of newly synthesized interferon-gamma (IFN-gamma) in anti CD3-stimulated lymphocytes treated with glycoprotein processing inhibitors. AB - As described previously (Kosuge T., Toyoshima S., Biol. Pharm. Bull., 23, 1-5 (2000)), inhibitors of the glycoprotein processing enzymes, glucosidase I and II, induce decreased secretion of interferon-gamma (IFN-gamma) into culture supernatants of anti CD3-stimulated lymphocytes, and in the present study the mechanism has been investigated in further detail. The processing inhibitors did not affect intracellular levels of IFN-gamma but enhanced the degradation of newly synthesized IFN-gamma in anti CD3-stimulated lymphocytes. Furthermore, since the stability of N-glycosylated proteins is known to be regulated by lectin family chaperones, such as calnexin, a type I transmembrane protein located in the endoplasmic reticulum (ER), and calreticulin, a soluble protein in the ER lumen, the effect of the processing inhibitors on the interaction of IFN-gamma with calnexin and calreticulin was investigated. It was found that IFN-gamma formed complexes with calnexin and calreticulin in anti CD3-stimulated lymphocytes. Total binding of IFN-gamma to calnexin was not affected but that to calreticulin was increased in anti CD3-stimulated lymphocytes treated with the processing inhibitors. However, binding of newly synthesized IFN-gamma to calreticulin was decreased in the lymphocytes under the same conditions as above. These results suggest that these glycoprotein processing inhibitors block the release of IFN-gamma from already formed calreticulin complexes, which prevents the binding of newly synthesized IFN-gamma to calreticulin and results in the enhancement of IFN-gamma degradation. PMID- 10823662 TI - Stimulatory release of hepatic lipase activity from rat hepatocytes by ruthenium red. AB - Ruthenium Red (RuR; ruthenium oxychloride ammoniated) stimulated the release of hepatic lipase (HTGL) activity from primary cultured rat hepatocytes into medium in a time- and dose-dependent manner. The RuR-stimulated release of HTGL activity was suppressed by tyrosine kinase (TK) inhibitors (ST-638 and biochanin A). The activity of partially purified TK preparation from hepatocytes was found to be increased by incubation with RuR. In addition, treatment of the hepatocytes with H-89, a potent inhibitor of cAMP-dependent protein kinase (PKA), decreased the stimulatory release of HTGL activity by RuR. Moreover, cAMP content in RuR incubated hepatocytes was rapidly increased, and activation of PKA was observed. The RuR-stimulated release of HTGL activity is also inhibited by uncouplers and glycosylation inhibitors. In addition, incorporation of [3H]leucine into protein was increased in the present of RuR. Under marked inhibition of protein synthesis by cycloheximide, RuR still showed a full effect on the release of HTGL activity. These results suggest that RuR stimulates the release of HTGL activity through mechanisms of action involving TK- and PKA-activating pathways, which require a metabolic energy-sensitive process rather than elevation of enzyme molecule synthesis. PMID- 10823663 TI - Gastric anti-ulcer activity of several alpha,beta-unsaturated carbonyl compounds in rats. AB - The gastric cytoprotective activity of several molecules containing an alpha,beta unsaturated carbonyl system is reported. We attributed this gastroprotective activity to the presence of a non-hindered Michael acceptor in the molecules assayed and suggested that the mechanism of protection would involve, at least in part, a nucleophilic attack of the sulphydryl group of the gastric mucosa to the beta carbon of the Michael acceptors of the compounds assayed. PMID- 10823664 TI - Study of the in vivo and in vitro cardiovascular effects of four new analogues of ketanserin: implication of 5-HT2A and alpha1 adrenergic antagonism in their hypotensive effect. AB - The in vivo and in vitro cardiovascular effects of the novel 5-HT2A/alpha1/H1 antagonist ketanserin analogues QF 0303B, QF 0307B, QF 0311B, QF 0313B were studied in anaesthetized normotensive rats (ANR) and in isolated rubbed rat aorta (IRRA). In ANR, 0.2 mg x kg(-1) i.v. of each compound produced a rapid, remarkable but short-lasting fall in mean arterial blood pressure (MAP) accompanied by bradycardia. All compounds significantly modified the pressor effects induced by 5-hydroxytryptamine (5-HT) and noradrenaline (NA). In IRRA, the compounds inhibited NA- and 5-HT-induced contractions in a competitive fashion. Furthermore, the analogues displayed lower H1-antagonist activity than ketanserin. Compounds tested showed low 5-HT2B affinity and no activity at muscarinic, nicotinic, or 5-HT3 receptors, nor any marked ability to produce smooth muscle relaxation via calcium entry blockade. There is a significant correlation between hypotension reached and inhibition of the 5-HT-induced pressor responses (but not for NA). A certain degree of correlation was observed between hypotensive effect endurance vs. alpha1-adrenoceptor blockade (but not for serotonin). These results indicate that in this series the brief hypotensive activity in ANR is attributed to a 5-HT1A receptor blockade and the duration of the effect is better attributed to an alpha1 adrenoceptor blockade. PMID- 10823665 TI - Experimental allergic conjunctivitis in guinea pigs induced by Japanese cedar pollen. AB - We report a new experimental allergic conjunctivitis with Japanese cedar pollen as antigen in guinea pigs, and the immunological characteristics of this model were also elucidated. Allergic conjunctivitis was developed by immunization in guinea pigs with a mixture containing Japanese cedar pollen and killed Bordetella pertussis. When local application of Japanese cedar pollen suspension 14 d after systemic immunization was performed every 3d, remarkable conjunctivitis was observed from 20 to 35 d. Increase in vascular permeability and decrease in histamine contents of the conjunctiva were also observed after local application of antigen. Passive cutaneous anaphylactic (PCA) reactions revealed that both IgG and IgE-rich antibodies were produced in this model. Chlorpheniramine, ketotifen and levocabastine were effective in inhibiting cedar pollen-induced conjunctivitis. Although a high concentration was needed, tranilast and amlexanox also showed significant inhibition of conjunctivitis induced by cedar pollen. PMID- 10823666 TI - Fluvastatin, an HMG-CoA reductase inhibitor, protects LDL from oxidative modification in hypercholesterolemic rabbits. AB - The antioxidative effect of fluvastatin sodium (fluvastatin) on low-density lipoprotein (LDL) was evaluated in vivo and in vitro. Since ex vivo measurement of the LDL oxidizability is reported to reflect the response of the atherosclerotic process, LDL isolated from rabbits fed a high cholesterol diet for 4 weeks with or without fluvastatin, pravastatin or alpha-tocopherol administration was oxidized by copper ions to estimate conjugated diene formation. Fluvastatin but not pravastatin significantly prolonged the lag time of LDL oxidized by copper ions ex vivo without affecting plasma cholesterol levels at a dose of 3 mg/kg after four weeks of treatment. Alpha-tocopherol treated rabbits showed dramatically elongated LDL oxidation lag time at a dose of 150 mg/kg. In order to assess the mechanism, the content of alpha-tocopherol, a major endogenous antioxidant in LDL was measured, and we found that only LDL isolated from alpha-tocopherol-treated rabbits contained a significantly larger amount of alpha-tocopherol than that from high cholesterol control rabbits. To elucidate the mechanism further, the effect of fluvastatin on conjugated diene formation during copper-induced LDL oxidation in vitro was studied. Fluvastatin not only prolonged lag time, but also suppressed the rate of LDL oxidation, both in a dose dependent manner above 1 microM, while pravastatin showed no effect. These results suggest the direct antioxidative effect of fluvastatin on LDL oxidation in vivo. Since oxidation of LDL is an important step in the initiation and progression of atherosclerosis, fluvastatin may reduce the risk of this condition not only by lowering plasma cholesterol but also by protecting LDL from oxidation. PMID- 10823667 TI - Nitric oxide inhibits dioxin action for the stimulation of Cyp1a1 promoter activity. AB - Since it is known that hypoxia increases inducible nitric oxide synthase (iNOS) gene expression through the hypoxia responsive element, it was hypothesized that nitric oxide could be a mediator of hypoxia to inhibit Cyp1a1 promoter activity. In order to test this hypothesis, we have undertaken a study to examine the effects of hypoxia and nitric oxide on Cyp1a1 promoter activity in Hepa I cells. Mouse Cyp1a1 5' flanking DNA, 1.6kb, was cloned into pGL3 expression vector in order to construct pmCyp1a1-Luc. Hepa I cells were transfected with pmCyp1a1-Luc and were treated with 10(-9)M 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in the presence or absence of various hypoxic agents such as 10(-6)-10(-4) cobalt chloride or 10(-6)-10(-4)M picolinic acid or 10(-6)-10(-4) M desferrioxamine. The luciferase activity of the reporter gene was measured from pmCyp1a1-Luc transfected Hepa I cell lysate which contains 2 microg total protein using luciferin as a substrate. Hypoxic agents such as cobalt chloride, picolinic acid, and desferrioxamine showed inhibition of luciferase activity that was induced by 10(-9) M TCDD treatment in a dose dependent manner. Concomitant treatment of 1 mM N(G)-nitro-l-arginine with 10(-6)-10(-4) M cobalt chloride or 10(-6)-10(-4) M desferrioxamine or 10(-6)-10(-4) M picolinic acid or 10(-6)-10(-4) M sodium nitroprusside recovered luciferase activity from the TCDD induced luciferase activity that was inhibited by hypoxic agents. These data demonstrated that nitric oxide might be a mediator of iron chelating agents and hypoxic agents to inhibit dioxin induced Cyp1a1 promoter activity in Hepa I cells. PMID- 10823668 TI - Sodium fluoride increases intracellular calcium in rat renal epithelial cell line NRK-52E. AB - In our previous experiment using rats, fluoride was reported to cause renal calcification, whose mechanism was deduced to be due to an increase in parathyroid hormone (PTH) secretion. However fluoride-induced renal calcification that was independent of PTH has not been understood well in the nephron of fluoride-treated animals. Thus, we examined the effect of sodium fluoride on intracellular calcium mobilization in a normal rat kidney epithelial cell line (NRK-52E cells). The calcium accumulation was found to be remarkably increased by the addition of sodium fluoride (NaF). The elevation of [Ca2+]i was demonstrated to be due to calcium entry through nifedipine-sensitive calcium channels. In addition, fluoride activates phospholipase C, but inositol 1,4,5-triphosphate (IP3) didn't induce Ca2+ release from the endoplasmic reticulum (ER). Moreover, fluoride alone was deduced to enhance the activity of ER-type Ca2+-ATPase. Finally, on the mechanism of fluoride-induced calcium accumulation in NRK-52E cells, fluoride may activate phospholipase C to generate IP3 and diacylglycerol, and these increases can be elucidated to induce calcium entry through dihydropiridine-sensitive calcium channels. Moreover, fluoride was found to stimulate calcium accumulation through ER-type Ca2+-ATPase into the endoplasmic reticulum. The elevation of ER-type Ca2+-ATPase activity by fluoride was elucidated to operate as a regulatory system to protect against abnormally higher increases in cytosolic calcium concentration via an increase of calcium influx into the endoplasmic reticulum. PMID- 10823669 TI - Authentication of an animal crude drug, Zaocys, by diagnostic PCR. AB - A pair of diagnostic primers for distinguishing the Chinese crude drug Zaocys (Zaocys dhumandes) from its substitutes was designed based on the sequence data of the original animal of the drug and substitutes. Total DNAs were extracted from genuine crude drug and 5 of its substitutes, as well as from 12 species of original animal of the snake crude drug. Diagnostic PCRs were performed using the primers with these total DNAs as a template, annealing at 60-65 degrees C. Positive amplifications were obtained from all DNA templates of Zaocys, whereas negative amplifications were obtained from that of others. The results indicate that Zaocys samples could be definitely distinguished from its substitutes by diagnostic PCR, and no incorrect discrimination was found under the same reaction conditions. The advantages of the method in the authentication of crude drugs are also discussed in the present paper. PMID- 10823670 TI - Identification of the crude drug atractylodes rhizome (Byaku-jutsu) and atractylodes lancea rhizome (So-jutsu) using chloroplast TrnK sequence as a molecular marker. AB - Novel methods for molecular authentication of Atractylodes-derived crude drugs (Jutsu) were established based on PCR-restriction fragment length polymorphism (RFLP) and direct sequencing of chloroplast trnK. Two regions inside the chloroplast trnK were selected as molecular markers for identification and discrimination of Atractylodes Rhizome (Byaku-jutsu) and Atractylodes Lancea Rhizome (So-jutsu). The Region 1 fragment (260 bp) amplified from So-jutsu and Wa byaku-jutsu (Atractylodes Rhizome derived from A. japonica) gave 2 bands of 180 bp and 80 bp on agarose gel electrophoresis after digestion with a restriction endonuclease HinfI, whereas the fragment amplified from Kara-byaku-jutsu (Atractylodes Rhizome derived from A. ovata) remained undigested, which allowed unambiguous identification of Kara-byaku-jutsu. By direct sequencing of Region 2 (436 bp) and comparison of the nucleotide sequence data sets we could not only discriminate Byaku-jutsu and So-jutsu but also identify the original plant species of each crude drug specimen. A simple and reliable protocol for rapid preparation of DNA suitable for PCR from as little as 1 mg of Atractylodes derived crude drugs was also described. PMID- 10823671 TI - Isolation of the antiulcer compound in essential oil from the leaves of Cryptomeria japonica. AB - Essential oil from the leaves of Tateyamasugi (Cryptomeria japonica) exhibited strong inhibitory activity on ulceration induced by HCl/ethanol, HCl/aspirin, water-immersion stress and pylorus-ligation. We separated the antiulcer compounds from cedar essential oil by use of distillation and chromatography. As a result, terpinen-4-ol, a monoterpene, and elemol, a sesquiterpene, were isolated as active compounds. The antiulcer activity of the former was more potent than that of the latter. Terpinen-4-ol was a mixture of optical isomers and each possessed potent antiulcer activity. Secretion of gastric juice and output of acid and pepsin activity were lowered by terpinen-4-ol. These results suggest that terpinen-4-ol isolated from cedar essential oil could be a valuable antiulcer agent. PMID- 10823672 TI - Inhibitory effects of methanol extracts of herbal medicines on substance P induced itch-scratch response. AB - In a search for new anti-pruritic drugs we screened methanol extracts of 33 herbal medicines which have been used for cutaneous diseases for their antipruritic activity using substance P (SP) as a pruritogen in mice. When administered perorally 30 min before SP injection, methanol extracts of 6 of these herbal medicines, the root of Scrophularia ningpoensis Hemsl., the root of Patrinia villosa (Thunb.) Juss, the fruit of Forsythia suspenna Vahl, the rhizome of Cimicifuga dahurica (Turcz.) Maxim., the aerial part of Schizonepeta tenuifolia Briq. and the fruit of Cnidium monnieri (L.) Cuss, inhibited SP induced itch-scratch response at a dose of 200 mg/kg with-out affecting locomotor activity. Dose dependence of these 6 extracts (50-500 mg/kg) was investigated and all of them inhibited SP-induced itch-scratch response, with extracts from Scrophularia ningpoensis, Schizonepeta tenuifolia and Cnidium monnieri showing particularly significant inhibition. The results suggest that these 6 methanol extracts have inhibitory activity against SP-induced itching. PMID- 10823673 TI - Phylogenetic relationship of six Glycyrrhiza species based on rbcL sequences and chemical constituents. AB - The nucleotide sequences of ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit gene (rbcL) of Glycyrrhiza glabra, G. uralensis, G. inflata, G. echinata, G. macedonica and G. pallidiflora have been determined to construct their phylogenetic tree. Based on these sequences, the six Glycyrrhiza species were divided into two groups: three, G. glabra, G. uralensis, and G. inflata, which produce glycyrrhizin as a major saponin, and the others, G. echinata, G. macedonica and G. pallidiflora, which produce macedonoside C as a major saponin. Among the three glycyrrhizin-producing species, only two nucleotide substitutions were observed between the rbcL sequences of G. glabra and G. uralensis, and the sequence of G. uralensis was identical to that of G. inflata, indicating that G. uralensis and G. inflata are closely related. Among the three macedonoside C producing species, only one nucleotide substitution was observed between those of G. echinata and G. macedonica, indicating that these two species are also closely related. PMID- 10823674 TI - Effects of Aconitum alkaloid kobusine and pseudokobusine derivatives on cutaneous blood flow in mice; II. AB - Aconitum alkaloids of the C20-diterpenoid type, kobusine (1) and pseudokobusine (2), their anisoyl, veratroyl, p-nitrobenzoyl, nicotinoyl or pivaloyl derivatives, and dehydrokobusine and N,6-seco-6-dehydropseudokobusine derivatives were examined for their peripheral vaso-activities by laser-flowmetrical measurement of the cutaneous blood flow in the hind foot of mice after intravenous administration. Kobusine 15-anisoate (4), 11-veratroate (5), 15 veratroate (6), 11-pivaloate (9) and 15-pivaloate (10) were significantly effective at a low dose of 0.5 or 0.05 mg/kg. Pseudokobusine derivatives were all active at 1, 0.5 or 0.05 mg/kg, and the effects of pseudokobusine 15-anisoate (13), 15-veratroate (16) and 15-p-nitrobenzoate (19) at 0.1 mg/kg were remarkable. Yesoline (26) and alkaloid (28) were significantly effective at a low dose of 1 mg/kg, whereas yesonine (25) and N-acetyl-N,6-seco-6 dehydropseudokobusine (27) were inactive. Dehydrokobusine derivatives (29, 30) were significantly effective at a low dose of 0.5 or 0.1 mg/kg. It is thought that the hydroxyl groups of alkaloids, especially a free OH group of 2 at C-6, are important for action on the peripheral vasculature leading to dilatation, and the results indicated that esterification of the hydroxyl group at C-15 with either anisoate, veratroate orp-nitrobenzoate may contribute to enhancement of the activity of the parent alkaloids. PMID- 10823675 TI - Preparation and in vivo ocular absorption studies of disulfiram solid dispersion. AB - Disulfiram, a dimer of diethyldithiocarbamate (DDC) which is a strong radical scavenger, is known to prevent cataract development. However, disulfiram is hardly absorbed from the cornea and its bioavailability is extremely low. In this study, we attempted to prepare disulfiram solid dispersion for the improvement of ocular bioavailability. Solid dispersions of disulfiram were prepared by either an evaporation method or a spray-drying method, using polyvinylpyrrolidone (PVP) as a carrier. Preparations were analyzed by scanning electron microscopy, powder X-ray diffractometry and differential scanning calorimetry, and confirmed to be a solid dispersion. The particle size of the solid dispersion prepared by the spray drying method was smaller than the preparation by the evaporation method (spray drying: 3.3+/-0.04 microm, evaporation: 34.3+/-18.0 microm). An in vivo ocular absorption experiment was conducted by instilling solid dispersions to rabbit eye and measuring the DDC in the aqueous humor. After instillation of disulfiram and PVP physical mixture, DDC was not detected in the aqueous humor. On the other hand, DDC appeared in the aqueous humor after the instillation of a solid dispersion. Maximal concentration and the area under the aqueous humor concentration-time curve were greater in the solid dispersion prepared by the spray-drying method than the preparation by the evaporation method. Disulfiram solid dispersion, especially prepared by the spray-drying method, improved ocular bioavailability. PMID- 10823676 TI - Distribution characteristics of carboxymethylpullulan-peptide-doxorubicin conjugates in tumor-bearing rats: different sequence of peptide spacers and doxorubicin contents. AB - Plasma and tissue distribution of conjugates of carboxymethylpullulan (CMPul) and doxorubicin (DXR), either bound directly or through three types of tetrapeptide spacers, was studied after intravenous injection to rats bearing Walker 256 carcinosarcoma and compared with that of DXR. In contrast to DXR, each conjugate retained high levels of DXR in the conjugated form in plasma and displayed high accumulation in the tumor at 6 h after the administration. Disposition characteristics of [3H]CMPul in rats bearing Walker 256 carcinosarcoma indicate that pullulan, which had molecular weight over 50 kDa, is a suitable macromolecular carrier for tumor targeting in cancer chemotherapy by carboxymethylation. We find that the in vivo antitumor effect of the conjugates depends on the tumor AUC of free DXR released from the conjugates. CMPul-DXR conjugates were also distributed in the reticuloendothelial organs, such as liver, spleen and bone marrow; however, the tissue concentrations of the conjugates in the heart, lung and muscle were lower than those of DXR. We next investigated the effect of the DXR contents of CMPul-DXR conjugates on their body distribution in rats bearing Walker 256. The half life of CMPul-DXR conjugates in plasma were shorter and the conjugates had greater accumulation in the reticuloendothelial system, while they showed lower concentrations in the tumor with increasing DXR contents. Antitumor activity of CMPul-DXR conjugates were reduced and the lethal toxicities of CMPul-DXR conjugates were amplified with increasing DXR contents. PMID- 10823677 TI - In vitro/in vivo characterization of a tramadol HCl depot system composed of monoolein and water. AB - Various monoolein-water systems containing tramadol HCl, a potent analgesic, were formulated to obtain sustained-release dosage forms which could be administered by subcutaneous, intramuscular or intrathecal injections. They were examined for their in vitro drug-release profiles and in vivo analgesic properties in rats in a 14 h period following intramuscular administration. In order to obtain a lower viscosity, we have substituted a part of monoolein by oleic acid and phospholipids. Both binary (monoolein-water) and quaternary (oleic acid phospholipid-monoolein-water) formulations exhibited controlled drug-release profiles which were accelerated by surfactant adjunction. This surfactant action was probably due to structural changes in the lipid arrangement and was much more pronounced for the modified formulations. According to the results obtained in vitro, formulations with slower drug release (i.e. the native formulation and the modified one without surfactant) were selected for assessment of their in vivo properties. Both formulations demonstrated prolonged analgesic activities in the rat tail flick test manifested by stable pain relief during more than 10 h compared with the 3 to 4 h analgesia obtained with the commercially available tramadol HCl solution. The sustained-release capabilities were evaluated by using a modified half value duration (HVD) ratio and all sustained-released formulations exhibited a HVD ratio equal or superior to 3.9. PMID- 10823678 TI - Iontophoretic transdermal delivery of ketoprofen: novel method for the evaluation of plasma drug concentration in cutaneous vein. AB - The objective of our study is to establish a novel method for the in vivo evaluation of transdermal delivery. In this study, cathodal iontophoresis of ketoprofen, a nonsteroidal anti-inflammatory drug, was performed in the thoracic area of rats at a constant direct current, and blood samples were collected from cutaneous vein passing through the thoracic part of the body. After the iontophoresis, the plasma ketoprofen concentration in cutaneous vein ipsilateral to the application site was significantly higher than that in systemic vein. On the other hand, the plasma concentration in cutaneous vein contralateral to the application site was not significantly different from that in systemic vein. A comparison of the time-course curves demonstrated that, for the duration of iontophoresis, the plasma ketoprofen concentration in cutaneous vein ipsilateral to the application site increased with the amount of ketoprofen absorbed in the skin. These results suggest that the plasma concentration in the cutaneous vein ipsilateral to the application site is related with the transfer of drug from skin to cutaneous blood circulation. Therefore, the measurement of plasma concentration in cutaneous vein close to the application site would allow us to directly quantify the local behavior of iontophoretic transdermal absorption. PMID- 10823679 TI - In vitro release of SM-11355, cis[((1R,2R)-1,2-cyclohexanediamine N,N')bis(myristato)] platinum(II) suspended in lipiodol. AB - SM-11355, cis[((1R,2R)-1,2-cyclohexanediamine-N,N')bis(myristato)] platinum(II) is a lipophilic platinum complex. SM-11355 suspended in Lipiodol (SM 11355/Lipiodol) was previously shown to have antitumor effects against rat hepatic tumors after intra-arterial administration. In the present study, the in vitro release of platinum compounds from SM-11355/Lipiodol was examined. A test tube containing 10 ml of saline and 1 ml of SM-11355/Lipiodol was rotated at 5 rpm in a vertical orientation. The platinum concentration in saline gradually increased for 28 d. From HPLC analysis, cyclohexane-1,2-diamineplatinum(II) dichloride (DPC) and cyclohexane-1,2-diamineplatinum(II) chloroiodide (DPCI) were detected in the saline, and the sum of these two compounds was equivalent to the total platinum amount in the saline determined by atomic absorption spectrophotometry at days 21 and 28. DPC showed significant growth inhibitory activities, with IC50 values of 0.1-0.7 nmol/ml in rat hepatoma AH-109A cells and 5 human tumor cell lines, as effective as cisplatin. These findings suggest that SM-11355/Lipiodol exerts antitumor effects by releasing active platinum compounds, and that DPC is one of the candidates of the active compounds. PMID- 10823680 TI - In vivo electron spin resonance assessment of decay constant of nitroxyl radical in selenium-deficient rat. AB - In vivo reducing capacity and cytosolic glutathione peroxidase (GSH-Px) activity in liver homogenate were evaluated in 6 weeks old Se-deficient and normal rats. GSH-Px was significantly lower in Se-deficient rats than in normal rats. In vivo reducing capacity in head and liver parts, estimated from in vivo signal decay of a nitroxyl spin probe using a low frequency (300 MHz) ESR spectrometer, was significantly decreased in Se-deficient rats, suggesting a decrease of antioxidant capacity in Se-deficient rats. PMID- 10823681 TI - Phytogrowth-Inhibitory activities of beta-dolabrin and gamma-thujaplicin, hinokitiol-related compounds and constituents of Thujopsis dolabrata Sieb. et Zucc. var hondai Makino. AB - Beta-dolabrin and gamma-thujaplicin isolated from Thujopsis dolabrata Sieb. et Zucc. var hondai Makino, like hinokitiol, showed strong phytogrowth-inhibitory activities, and their growth-inhibitory activities were as high as that of sodium 2,4-dichlorophenoxyacetate used as a positive control. In particular, the phytogrowth-inhibitory activity of gamma-thujaplicin was strong and it completely inhibited the germination of this seed of Brassica campestris L. subsp. rapa Hook f. et Anders at the concentration of 30 ppm. Both compounds exhibited inhibitory activities on B. campestris L. subsp. rapa Hook f. et Anders and Sesamum indicum Linne, even at the low concentration of 10 ppm. At 7 d after treatment with beta dolabrin and gamma-thujaplicin, the amount of chlorophyll in the cotyledons of B. campestris L. subsp. rapa Hook f. et Anders treated with both compounds was greatly decreased as compared with the control. The findings indicate that the phytogrowth-inhibitory action might be a common biological activity of hinokitiol related compounds, suggesting that at least a part of their phytogrowth inhibitory actions seems to be related to a decrease in chlorophyll content. PMID- 10823682 TI - Cloning and characterization of the ddc homolog encoding L-2,4-diaminobutyrate decarboxylase in Enterobacter aerogenes. AB - L-2,4-diaminobutyrate decarboxylase (DABA DC) catalyzes the formation of 1,3 diaminopropane (DAP) from DABA. In the present study, the ddc gene encoding DABA DC from Enterobacter aerogenes ATCC 13048 was cloned and characterized. Determination of the nucleotide sequence revealed an open reading frame of 1470 bp encoding a 53659-Da protein of 490 amino acids, whose deduced NH2-terminal sequence was identical to that of purified DABA DC from E. aerogenes. The deduced amino acid sequence was highly similar to those of Acinetobacter baumannii and Haemophilus influenzae DABA DCs encoded by the ddc genes. The lysine-307 of the E. aerogenes DABA DC was identified as the pyridoxal 5'-phosphate binding residue by site-directed mutagenesis. Furthermore, PCR analysis revealed the distribution of E. aerogenes ddc homologs in some other species of Enterobacteriaceae. Such a relatively wide occurrence of the ddc homologs implies biological significance of DABA DC and its product DAP. PMID- 10823683 TI - Evaluation of analgesic and anti-inflammatory activity of novel beta-lactam monocyclic compounds. AB - We have examined the in vivo anti-inflammatory and analgesic activity of a new series of monocyclic beta-lactams (azetidinones), similar to others which have been demonstrated to be inhibitors of human leukocyte elastase (HLE), an enzyme involved in degradation processes of connective tissue. Our new compounds have been administered orally (15 mg/kg) to albino rats 30 min before injecting carrageenin in the plantar aponeurosis. Tested compounds have demonstrated a certain activity and stability to gastric hydrolysis, in particular two of them markedly reduced paw edema formation, even if slightly less effectively than indomethacin (reference compound, 5 mg/kg). To evaluate the analgesic activity we carried out the acetic acid writhing test, pretreating rats orally with our compounds 30 min before injecting the acid solution i.p. The same two molecules which showed the anti-inflammatory activity demonstrated a very light analgesic activity. These results suggest the possibility of carrying out further studies, particularly in vitro, on the mechanism of action of our compounds, mechanism which could be the HLE inhibition. PMID- 10823684 TI - Amitriptyline-induced constipation in cynomolgus monkeys is beneficial for the evaluation of laxative efficacy. AB - In an attempt to create an animal model of constipation in monkeys, amitriptyline was administered to cynomolgus monkeys at doses of 10-160 mg/kg body weight via a nasogastric tube. Normal control monkeys excreted feces frequently throughout the day. Monkeys treated with amitriptyline at doses of 10-40 mg/kg showed delays in feces excretion. The 60 mg/kg treated monkeys for the most part did not excrete feces during the 24 h after amitriptyline administration. The 80 and 120 mg/kg treated monkeys did not excrete feces until 24 h from administration of amitriptyline, and also showed prolonged crouching and lethargy. On the other hand, 160 mg/kg treated monkeys died within 24 h after administration. We therefore felt that the optimal dose for creating constipation in the monkeys was 60 mg/kg. We tested the appropriateness of this amitriptyline-induced constipated monkey model by observing the effects of a new laxative, the herbal medicine ND 10 and the commercially available laxative bisacodyl. Control monkeys (those not receiving ND-10 or bisacodyl) treated with 60 mg/kg amitriptyline did not excrete feces up to 32 h after amitriptyline administration in 2 of 3 monkeys. However, all monkeys treated with one tablet of ND-10 excreted feces. Also, in 4 monkeys administrated with bisacodyl, 3 excreted feces. In this study, we confirmed that constipation can be caused in cynomolgus monkeys by oral administration of amitriptyline. This model may also be useful for the evaluation of laxatives. PMID- 10823685 TI - Saponins isolated from Allium chinense G. Don and antitumor-promoting activities of isoliquiritigenin and laxogenin from the same drug. AB - Investigation of the Chinese crude drug "Xiebai," the bulbs of Allium chinense G. Don (Liliaceae), led to the isolation of 2 saponins, xiebai-saponin I (laxogenin 3-O-beta-xylopyranosyl (1-->4)-[alpha-arabinopyranosyl (1-->6)-beta glucopyranoside) (1) and laxogenin 3-O-alpha-arabinopyranosyl (1-->6)-beta glucopyranoside (2), and the aglycone, laxogenin (3), together with 2 chalcones, isoliquiritigenin (4) and isoliquiritigenin-4-O-glucoside (5), and beta sitosterol glucoside (6). Compounds 1-5 were tested in vitro for their inhibitory effect on the 12-O-tetradecanoylphorbol-13-acetate (TPA)-stimulated 32Pi incorporation into phospholipids of HeLa cells. In addition to this, laxogenin (3) was proven to have an antitumor-promoting activity in a two-stage lung carcinogenesis experiment. PMID- 10823686 TI - Effects of sulfhydryl compounds on interleukin-1-induced vascular endothelial growth factor production in human synovial stromal cells. AB - We investigated the effects of various sulfhydryl compounds on interleukin-1 (IL 1)-induced vascular endothelial growth factor (VEGF) production in human synovial stromal cells (HSSC). HSSC stimulated by IL-1beta (100 ng/ml) produced VEGF and interleukin-6 (IL-6) in vitro. Monosulfhydryl compounds, N-acetylcysteine, D penicillamine, tiopronin and the bucillamine-like disulfhydryl compound, compound A scarcely affected VEGF or IL-6 production at concentrations of 10(-5) and 10( 4) M. However, the disulfhydryl compound, bucillamine inhibited VEGF production but not IL-6 production at concentrations of 10(-5) and 10(-4) M. These results suggest that bucillamine may be a selective inhibitor of IL-1-induced VEGF production in HSSC, and that inhibition of VEGF production may require not only SH groups but also a specific chemical structure. PMID- 10823687 TI - Sauchinone, a lignan from Saururus chinensis, attenuates CCl4-induced toxicity in primary cultures of rat hepatocytes. AB - We used primary cultures of rat hepatocytes injured by the hepatotoxin, carbon tetrachloride (CCl4), as a test system to screen for hepatoprotective compounds from natural products. Sauchinone was isolated from the aerial parts of Saururus chinensis (Saururaceae) by this method. At a concentration of 50 microM, sauchinone significantly reduced the release into the culture medium of glutamic pyruvic transaminase from CCl4-damaged cultures of rat hepatocytes. It has been determined that glutathione, superoxide dismutase and glutathione peroxidase all play important roles in the cellular defense against oxidative stress. Sauchinone appeared to protect primary cultured rat hepatocytes exposed to CCl4 from significant drops in the levels of each of these three specific markers, respectively. Sauchinone also seemed to ameliorate lipid peroxidation as demonstrated by a reduction in the production of the oxidized lipid byproduct, malondialdehyde. These results suggest that sauchinone may exert hepatoprotective activity through antioxidant activity. PMID- 10823688 TI - Inhibitory effect of azole antifungal agents on the glucuronidation of lorazepam using rabbit liver microsomes in vitro. AB - Azole antifungal agents (azoles) have inhibitory effects on the cytochrome P450. However, the effect of azoles on conjugative metabolism has not been given much attention. Lorazepam (LZP), a benzodiazepine sedative agent, is known to be metabolized by uridine 5'-diphosphate (UDP)-glucuronyltransferase. Herein we report investigation of the effect of azoles on the enzyme-kinetics of glucuronidation of lorazepam using rabbit liver microsomes in vitro. The Km and Vmax for LZP glucuronidation were determined to be 0.26+/-0.08 mM and 1.25+/-0.21 nmol/min/mg protein, respectively, when evaluated in the presence of a detergent 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate (CHAPS) (0.8 mg/mg protein). Azoles fluconazole, miconazole, and ketoconazole competitively inhibited the glucuronidation of LZP, with Ki values of 7.17+/-4.78 mM, 0.17+/ 0.08 mM, and 0.092+/-0.026 mM, respectively. These results are comparable to the previously reported Ki values of azoles with zidovudine (AZT) glucuronidation (1.4, 0.18, and 0.08 mM for fluconazole, miconazole, and ketoconazole, respectively) [Sampol et al., Br. J. Clin. Pharmacol., 40, 83-86, 1995]. Therefore, in order to avoid possible side effects of LZP, the concomitant administration of LZP and azoles should be carefully evaluated. PMID- 10823689 TI - Solubilized cell wall beta-glucan, CSBG, is an epitope of Candida immune mice. AB - Antibody to beta-glucan is generally difficult to produce in mice. We have recently developed a protocol to obtain a soluble Candida spp. beta-(1-->3)-D glucan (CSBG) by sodium hypochlorite (NaClO) oxidation and subsequent dimethyl sulfoxide (Me2SO) extraction. CSBG is composed mainly of beta-(1-->3) and beta-(1 ->6)-glucosidic linkages with a small amount of branch. In this paper, mice were immunized with Candida albicans and the specificity of the resulting sera to CSBG was examined by ELISA. Using CSBG coated plate, sera of the Candida immune mice showed higher reactivity than non-immune, normal mice and the reactivity was neutralized by adding soluble CSBG as a competitor. However, the reactivity could not be neutralized by a beta-(1-->6) branched beta-(1-->3)-glucan, grifolan. Similar specificity of the sera was obtained by commercially available beta glucan particle, zymosan or zymocel, immune mice. These facts strongly suggested that CSBG included epitopes of the specific antibody in Candida immune mice. PMID- 10823690 TI - Six new ursane- and oleanane-type triterpenes from the aerial roots of Ficus microcarpa. AB - Four new ursane-type triterpenes, 3beta-acetoxy-11alpha-methoxy-12-ursene (1), 3beta-acetoxy-11alpha-ethoxy-12-ursene (2), 3beta-acetoxy-11alpha-hydroperoxy-12 ursene (3), 3beta-hydroxy-11alpha-hydroperoxy-12-ursene (4), and two new oleanane type triterpenes, 3beta-acetoxy-11alpha-ethoxy-12-oleanene (5), 3beta-acetoxy 111alpha-hydroperoxy-12-oleanene (6), together with 3beta-acetoxy-11alpha-hydroxy 12-ursene (7), 3beta,11alpha-diacetoxy-12-ursene (8), 3beta-acetoxy-11alpha hydroxy-12-oleanene (9), were isolated from the aerial roots of Ficus microcarpa L. f. Their structures were elucidated by spectroscopic and chemical methods. PMID- 10823691 TI - Six podocarpane-type trinorditerpenes from the bark of Taiwania cryptomerioides. AB - Six podocarpane-type trinorditerpenes were isolated from the bark of Taiwania cryptomerioides. Their structures, 14-hydroxy-13-methoxy-8,11,13-podocarpatrien-7 one (1), 13-hydroxy-12-methoxy-8,11,13-podocarpatriene (2), 12-hydroxy-13-methoxy 8,11,13-podocarpatriene (3), 14-hydroxy-13-methoxy-8,11,13-podocarpatriene (4), 13-hydroxy-8,11,13-podocarpatriene (5), and 13,14-dihydroxy-8,11,13 podocarpatrien-7-one (6), were determined principally from spectral evidence. PMID- 10823692 TI - Determination of secnidazole in urine by adsorptive stripping voltammetry. AB - Cyclic voltammetry was used to explore the adsorption behavior of secnidazole on a hanging mercury drop electrode (HMDE). The effects of various operational parameters on the accumulation behavior of the adsorbed species were tested. Thus, a sensitive stripping voltammetry procedure for the determination of secnidazole with an adsorptive accumulation on the surface of HMDE has been developed. Measurements were taken by differential-pulse voltammetry after determination of the optimum conditions. The linear concentration range was 1 x 10(-8)-1 x 10(-7) s when using a 120 s preconcentration at -0.1 V vs. Ag/AgCl in acetate buffer of pH 4.0. The detection limit of secnidazole was 5 x 10(-9) M. The precision, expressed by the coefficient of variation, was 2.5% (n = 10) at a concentration of 1 x 10(-7) m. The method was successfully applied to the analysis of secnidazole in urine. PMID- 10823693 TI - Effect of polymethylene and phenylene linking groups on the DNA cleavage specificity of distamycin-linked hydroxamic acid-vanadyl complexes. AB - Two types of distamycin-linked hydroxamic acids (DHA), which contain various lengths of polymethylene chains (PM-DHA) and relatively rigid phenylene ones (Ph DHA), have been synthesized for the first time. Their DNA cleavage specificities were investigated by an end-labeled fragment cleavage experiment in the presence of vanadyl ion and hydrogen peroxide. The DNA cleavage by the PM-DHA x VO(II) complexes was shown to be very dependent on the length of the chain and the AT sequences. The tetramethylene DHA (1b) complex exhibited highly specific cleavage patterns flanking the 8 and 10 AT sites. Interestingly, the Ph-DHA complexes selectively cleaved the 5' end-labeled strand at the AT sites, but did not cleave the 3' end-labeled strand. The vanadyl complexing moieties and the local sequence conformation of the AT tract are suggested to contribute significantly to the DNA recognition of the PM-DHA x VO(II) complexes. PMID- 10823694 TI - Electron spin resonance studies of dipalmitoylphosphatidylcholine liposomes containing soybean-derived sterylglucoside. AB - The effects of soybean-derived sterylglucoside (SG) on the fluidity of liposomal membrane composed of dipalmitoylphosphatidylcholine (DPPC) were investigated compared with those of soybean-derived sterol (SS) and cholesterol (Ch) using an electron spin resonance spectrometer. Three kinds of liposomes were prepared in the molar ratio of DPPC/X=7/4, where X is SS, Ch or SG. The fluidity close to the polar head groups increased with an increase of temperature in the DPPC membrane containing SS, Ch and SG in the range 35 to 45 degrees C. Those near the hydrophobic end changed with an increase in temperature in liposomes containing SS, Ch and SG, which had a fluidizing effect on the DPPC membrane below the transition temperature (Tm, 41.9 degrees C) and a condensing effect over the Tm. The fluidizing effects of these compounds around 37 degrees C near the polar head group and the hydrophobic end increased in the following order: Ch < SG < or = SS and SS < Ch < SG, respectively. SG increased the fluidity of liposomal membrane dramatically above the Tm (35.4 degrees C). These results suggest that the high fluidity close to the hydrophobic end of the liposomal membranes around 37 degrees C, the decrease of Tm, and the sigmoidal nature of fluidity vs. temperature are important factors in the effectiveness of liposomes containing SG as a carrier of drugs. PMID- 10823695 TI - New cytotoxic butanolides from Lindera obtusiloba BLUME. AB - Three new butanolides, 2-(1-methoxy-11-dodecenyl)-penta-2,4-dien-4-olide (1), (2Z,3S,4S)-2-(11-dodecenylidene)-3-hydroxy-4-methylbutano lide (2) and (2E,3R,4R) 2-(11-dodecenylidene)-3-hydroxy-4-methoxy-4-methylbu tanolide (3), were isolated from the stems of Lindera obtusiloba BLUME. Their chemical structures were assigned by spectroscopic evidence. They exhibited cytotoxicity against cultured human tumor cell lines with their ED50 values ranging from 3.19 to 14.63 microg/ml. PMID- 10823696 TI - Modification of the physicochemical properties of minocycline hydrochloride ointment with cyclodextrines for optimum treatment of bedsore. AB - Modification to find the best physicochemical properties of minocycline hydrochloride ointment for optimum treatment of bedsore was investigated by coformulating various types of cyclodextrins (CyD) in the ointment base. It was found that the drug release rate from the ointment base was modified according to the preparation method of ointment base and the type of CyD admixed. The physicochemical properties, such as viscosity, elution volume, water absorption of ointment base were also modified by those factors. The mechanism of physicochemical modification with CyD was explained by the structural change of ointment base and the change of surface tension of emulsifying agent solution with the CyD. The stability of ointment was investigated by confirming the reproducibility of drug release rate after storage at ambient and cooled temperature conditions. In conclusion, a fused mixed ointment with beta-CyD was found to be preferable for treatment of bedsore, because of the improved drug release rate, lowered viscosity and increased elution volume of the resultant ointment. PMID- 10823697 TI - Synthesis and serotonin 2 (5-HT2) receptor antagonist activity of 5-aminoalkyl substituted pyrrolo[3,2-c]azepines and related compounds. AB - A series of 5-aminoalkylpyrrolo[3,2-c]azepine derivatives was synthesized and their serotonin 2 (5-HT2) receptor antagonist and antiplatelet aggregation activities were evaluated. 5-HT2 receptor antagonist activity was largely determined by the nature of the substituent at the 8-position as well as the aminoalkyl group at the 5-position of the pyrrolo[3,2-c]azepine ring. Compound 18a, 5-[3-[4-(4-fluorophenyl)piperazin-1-yl]propyl]-8-hydroxy-1-methyl- 1,4,5,6,7,8-hexahydropyrrolo[3,2-c]azepin-4-one, was recognized as having potent 5-HT2 receptor antagonist activity with weak alpha1 adrenoceptor blocking activity and no significant D2 receptor binding affinity, while the corresponding isomeric pyrrolo[3,4-c]azepine derivative (22) displayed only weak 5-HT2 receptor antagonist activity. After racemic 18a was resolved directly via diastereomeric salt formation, each enantiomer was evaluated precisely. The 5-HT2 receptor antagonist activity of 18a was found to reside primarily in (-)-18a (which was about 14-fold more potent than (+)-18a in isolated guinea pig arteries). Consequently, (S)-(-)-18a (SUN C5174) displayed the overall best profile with potent 5-HT2 receptor antagonist activity (pA2=8.98+/-0.06) and high selectivity versus other receptors. SUN C5174 showed a marked inhibitory effect on the platelet aggregation induced by serotonin in combination with collagen and adenosine diphosphate (ADP) in canine or human platelet-rich plasma (IC50=6.5 to 16 nM). Moreover, this compound significantly inhibited the mortality rate in mouse acute pulmonary thromboembolytic death induced by collagen and serotonin at oral doses of 0.3 mg/kg or higher. SUN C5174 is currently undergoing clinical evaluation. PMID- 10823698 TI - Benzaldehyde, 2-hydroxybenzoyl hydrazone derivatives as inhibitors of the corrosion of aluminium in hydrochloric acid. AB - The effect of benzaldehyde, 2-hydroxybenzoyl hydrazone derivatives on the corrosion of aluminium in hydrochloric acid has been investigated using thermometric and polarization techniques. The inhibitive efficiency ranking of these compounds from both techniques was found to be: 2>3>1>4. The inhibitors acted as mixed-type inhibitors but the cathode is more polarized. The relative inhibitive efficiency of these compounds has been explained on the basis of structure of the inhibitors and their mode of interaction at the surface. Results show that these additives are adsorbed on an aluminium surface according to the Langmuir isotherm. Polarization measurements indicated that the rate of corrosion of aluminium rapidly increases with temperature over the range 30-55 degrees C both in the absence and in the presence of inhibitors. Some thermodynamic data of the adsorption process are calculated and discussed. PMID- 10823699 TI - Lupin alkaloids from Chinese Maackia amurensis. AB - Two new alkaloids were isolated together with 16 known lupin alkaloids from the leaves and stems of Chinese Maackia amurensis. Their structures were determined by spectroscopic methods to be (-)-6alpha-methoxylupanine and (-)-5alpha-(12 cytisinylmethyl)-6alpha-hydroxylupan ine and identified by comparison with synthetic samples. The structures of lupin alkaloids were also related to the geographical distributions of the Maackia plants. PMID- 10823700 TI - Effects of aging on crystallization, dissolution and absorption characteristics of amorphous tolbutamide-2-hydroxypropyl-beta-cyclodextrin complex. AB - The effects of storage on the crystallization, dissolution and absorption of tolbutamide from amorphous tolbutamide-2-hydroxypropyl-beta-cyclodextrin (HP-beta CyD) complex were investigated, in comparison with those of polyvinylpyrrolidone (PVP) solid dispersion. The amorphous solid complex of tolbutamide with HP-beta CyD and the solid dispersion of tolbutamide with PVP were prepared by a spray drying method. During storage, a stable form of tolbutamide (form I) was crystallized from the amorphous PVP dispersion, whereas a metastable form of tolbutamide (form II) was crystallized from the HP-beta-CyD complex. The dissolution rate of tolbutamide from both HP-beta-CyD complex and PVP dispersion was significantly faster than that of tolbutamide alone. However, the dissolution rate from the PVP dispersion markedly decreased with storage, because of the formation of slow dissolving form I crystals. On the other hand, the dissolution rate from the HP-beta-CyD complex was only slightly decreased due to the formation of fast dissolving formII crystals. These in vitro dissolution characteristics were clearly reflected in the in vivo absorption of tolbutamide and the glucose plasma level after oral administration in dogs. The results suggested that HP-beta-CyD is useful not only for converting crystalline tolbutamide to an amorphous substance, but also for maintaining the fast dissolution rate of the drug over a long period. Furthermore, the crystallization of drugs from CyD complexes, with storage, seemed to be different from that involving polymer excipients such as PVP. PMID- 10823701 TI - Medicinal flowers. II. Inhibitors of nitric oxide production and absolute stereostructures of five new germacrane-type sesquiterpenes, kikkanols D, D monoacetate, E, F, and F monoacetate from the flowers of Chrysanthemum indicum L. AB - The methanolic extract and ethyl acetate-soluble portion from the flowers of Chrysanthemum indicum L., Chrysanthemi Indici Flos, were found to show inhibitory activity against nitric oxide (NO) production in lipopolysaccharide-activated macrophages. Five new germacrane-type sesquiterpenes, kikkanols D, D monoacetate, E, F, and F monoacetate, were isolated from the ethyl acetate-soluble portion. Their absolute stereostructures were elucidated on the basis of chemical and physicochemical evidence, which included application of the modified Mosher's method. The effects of fifteen principal components from the ethyl acetate soluble portion of this medicinal flower against NO production were examined and, among them, acetylenic compounds and flavonoids were found to show potent inhibitory activity. PMID- 10823702 TI - Three new sesquiterpene lactones from the pericarps of Illicium merrillianum. AB - Structures of three new sesquiterpene lactones 1-3, isolated from the pericarps of Illicium merrillianum, have been assigned as 14-O-benzoylfloridanolide, 2,10 epoxy-3-dehydroxypseudoanisatin and 7-O-methylpseudomajucin on the basis of spectroscopic data and chemical transformation. The structure of 2, having an ether linkage between C-2 and C-10, has been confirmed by X-ray crystallographic analysis. PMID- 10823703 TI - Application of liquid chromatography-atmospheric pressure chemical ionization mass spectrometry to the differentiation of stereoisomeric C19-norditerpenoid alkaloids. AB - High-performance liquid chromatography-atmospheric pressure chemical ionization mass spectrometry (HPLC-APCI-MS) was successfully applied to stereoisomeric C19 norditerpenoid alkaloids at position 1. APCI-MS allowed the easy and precise control of the energy deposition by varying the drift voltage. Comparison of the breakdown curves, observed by changing the potential difference between the first electrode and the second electrode of the APCI ion source, revealed stereochemical dependence of different fragmentations. The APCI spectra of alkaloids were predominantly the [M+H]+ ion and major fragment ion, corresponding to the [M+H-H2O]+ ion or the [M+H-CH3COOH]+ ion, and comparison of the spectra showed that the abundance of fragment ions was significantly higher for C-1 beta form alkaloids than for C-1 alpha-form alkaloids. The characteristic fragment ions were formed by the loss of a water, acetic acid or methanol molecule at position 8. The fragmentation mechanisms depending on the stereochemistry of the precursor ion could be discerned by recording the spectra in a deuterated solvent system of 0.05 M ammonium acetate in D2O-acetonitrile-tetrahydrofuran. Loss of D2O from the precursor ion gave the fragment ion. This result indicated that the proton of protonation was included in the leaving water molecule. The peak intensity ratio R=[M+H]+/[M+H-H2O]+ manifested the stereochemical differentiation of alkaloids at position 1. PMID- 10823704 TI - The cyclization reaction of ortho-ethynylbenzaldehyde derivatives into isoquinoline derivatives. AB - In order to elucidate the reaction mechanism of the cyclization between an ethynyl group and an imino group at the ortho-position on an aromatic ring to afford isoquinolines, reaction of 2-ethynylbenzaldehydes under various conditions was examined. It is concluded that reaction proceeds via an ionic process and the isoquinoline 4-hydrogen atom derives from the solvent. In addition, it was found that 2-ethynylbenzaldehyde O-methyloximes underwent cyclization in the presence of primary and secondary alcohols to give 3-substituted isoquinolines. PMID- 10823705 TI - Sustained-release phenylpropanolamine hydrochloride bilayer caplets containing the hydroxypropylmethylcellulose 2208 matrix. I. Formulation and dissolution characteristics. AB - The purpose of this study was to develop a new sustained-release phenylpropanolamine hydrochloride (PPA) bilayer caplets that consists of an immediate-release portion and a prolonged-release portion containing a hydroxypropylmethylcellulose 2208 (HPMC2208) matrix. Since PPA is a highly water soluble drug, incorporation of 60% HPMC2208 level in the matrix was required for giving the product a PPA-slow releasing property. Difference in the viscosity grade of HPMC2208 in the matrices did not greatly influence the PPA dissolution characteristics from the matrices. Therefore, we formulated the prolonged-release portion consisting of 10% PPA, 30% excipients, and 60% HPMC2208 (Metolose 90SH4000) into the sustained-release PPA bilayer caplets. The PPA dissolution characteristics from the formulated bilayer caplets showed the prolonged dissolution profile after rapid dissolution and was close to the targeted profile calculated from PPA pharmacokinetics study. The manufacturing methods of the prolonged-release portion and the filling order of the prolonged-release portion in bilayer compression did not significantly affect the PPA dissolution characteristics from the bilayer caplets. The PPA dissolution characteristics from the bilayer caplets was pH independent. Moreover, the PPA dissolution characteristics from the bilayer caplets was not affected by mechanical shear. The sustained-release PPA bilayer caplets is expected to present constant prolonged-release of PPA after rapid dissolution in vivo without dissolution change due to pH and mechanical shear. PMID- 10823706 TI - Sustained-release phenylpropanolamine hydrochloride bilayer caplets containing the hydroxypropylmethylcellulose 2208 matrix. II. Effects of filling order in bilayer compression and manufacturing method of the prolonged-release layer on compactibility of bilayer caplets. AB - The purpose of this study was to establish the manufacturing method of the formulated bilayer caplets containing the hydroxypropylmethylcellulose 2208 (HPMC2208) matrix without lamination. In manufacturing the bilayer caplets containing the HPMC2208 (Metolose 90SH4000) matrix, some bilayer caplets were cracked. We found that cracking of bilayer caplets is not the separation of two layers, but lamination of the prolonged-release layer. It was assumed that Metolose 90SH4000 causes lamination of the prolonged-release layer. Two factors, roller compaction pressure on dry granulation of the prolonged-release layer and filling order of the prolonged-release layer in bilayer compression, were related to lamination of bilayer caplets. The compactibility of the prolonged-release layer decrease with an increase in roller compaction pressure on dry granulation. The compactibility of the prolonged-release layer manufactured by direct compression is superior to that manufactured by dry granulation. The compactibility of the prolonged-release layer in the shape of the second layer, convexo-concave, is superior to that in the shape of the first layer, convexo convex. This is due to the fact that the density distribution inside the compact in the shape of convexo-concave was more uniform than that in the shape of convexo-convex. The manufacturing method of the formulated bilayer caplets having the prolonged-release layer whose Metolose 90SH4000 content is 60% without lamination is as follows: the prolonged-release layer manufactured by direct compression is fed as the second layer in bilayer compression. PMID- 10823707 TI - Synthesis of new synthons for organofluorine compounds from halothane containing sulfur functional groups. AB - To develop new synthons for the syntheses of organofluorine compounds, the treatment of Halothane, 2-bromo-2-chloro-1,1,1-trifluoroethane, (1) with 4 methylbenzenethiol (2) in the presence of sodium hydride gave 1-chloro-2,2,2 trifluoroethyl 4-methylphenyl sulfide (3), which was oxidized with m chloroperbenzoic acid (m-CPBA) to the corresponding sulfoxide (4) and sulfone (5). Reaction of 3 and 5 with allyltributyltin in the presence of 2,2' azobis(isobutyronitrile) (AIBN) gave 1-(trifluoromethyl)-3-butenyl compounds (9, 11). Sulfoxide 4 was decomposed in this condition. The treatment of 3 with allyltrimethylsilane in the presence of Lewis acids gave 1-(trifluoromethyl)-3 butenyl compounds (9) in good yield. This result suggests that 4-methylphenylthio substituent stabilizes the alpha-carbocation effectively, though the trifluoromethyl group destabilizes it strongly. Aromatic compounds similarly reacted with 3 in the presence of titanium(IV) chloride to give 2-aryl-1,1,1 trifluoro-2-(4-methylphenylthio)ethanes. Thus, sulfur compounds derived from Halothane were found to be useful new synthons for organofluorine compounds. PMID- 10823708 TI - Polyphenols from Eriobotrya japonica and their cytotoxicity against human oral tumor cell lines. AB - Three new flavonoid glycosides, together with 15 known flavonoids, have been isolated from the leaves of Eriobotrya japonica, and characterized as (2S)- and (2R)-naringenin 8-C-alpha-L-rhamnopyranosyl-(1-->2)-beta-D-glucopyranosides, and cinchonain Id 7-O-beta-D-glucopyranoside, respectively, based on spectral analyses including two dimensional (2D) NMR techniques. Higher proanthocyanidin fraction in the water-soluble portion of the extract was characterized as a procyanidin oligomer mixture mainly composed of undecameric procyanidin. These polyphenols have also been assessed for cytotoxic activity against two human oral tumor (human squamous cell carcinoma and human salivary gland tumor) cell lines. Selective cytotoxicity of the procyanidin oligomer between tumor and normal gingival fibroblast cells, and its possible mechanism, were also described. PMID- 10823709 TI - Synthesis and antifungal activities of R-102557 and related dioxane-triazole derivatives. AB - Novel triazole compounds with a dioxane ring were synthesized. Condensation of the diol precursor 10 with various aromatic aldehydes 11-13 under acidic conditions afforded a series of dioxane-triazole compounds 14-16. The antifungal activities of the compounds 14-16 were evaluated in vivo in mice infection models against Candida and Aspergillus species. High activities were seen for the derivatives with one or two double bond(s) and an aromatic ring substituted with an electron-withdrawing group in the side chain. Among the derivatives, R-102557 (16R: Ar=4-(2,2,3,3-tetrafluoropropoxy)phenyl) showed excellent in vivo activities against Candida, Aspergillus and Cryptococcus species. It also showed high tolerance in a preliminary toxicity study in rats. PMID- 10823710 TI - Intermediate state during the crystal transition in aspartame, studied with thermal analysis, solid-state NMR, and molecular dynamics simulation. AB - Aspartame (L-alpha-aspartyl-L-phenylalanine methyl ester) is a dipeptide sweetener about 200 times as sweet as sugar. It exists in crystal forms such as IA, IB, IIA, and IIB, which differ in crystal structure and in the degree of hydration. Among these, IIA is the most stable crystal form, and its crystal structure has been well determined (Hatada et al., J. Am. Chem. Soc., 107, 4279 4282 (1985)). To elucidate the structural factors of thermal stability in the IIA form of aspartame and to examine the physical process in the crystal transformation between the IIA and IIB forms, we performed a thermal analysis and solid-state NMR measurements. We found that a quasi-stable intermediate state exists in the transformation, and it has the same crystal lattice as the usual IIA form, despite the dehydration from 1/2 mol to 1/3 mol per 1 mol of aspartame. The results of the energy component analysis and the molecular dynamics simulation suggest that the entropic effect promotes the generation of the intermediate state, which is presumably caused by the evaporation of the water of crystallization and the increase of molecular motion in aspartame. Thus, the thermal stability of the IIA form is attributable to a structural property, i.e., the crystal lattice itself is retained during the above dehydration. Moreover, the molecular dynamics simulations suggest that the aspartame molecules have two kinds of conformational flexibility in the intermediate state. PMID- 10823711 TI - Stereocontrolled synthesis of piperidine-condensed tricyclic carbapenems (5 azatrinems) and their antibacterial activities. AB - Stereocontrolled synthesis of tricyclic carbapenem (5-azatrinem) derivatives 4, in which a piperidine ring is condensed to the carbapenem skeleton, was achieved. The pivotal tricyclic intermediate 2, allyl (8S,9R,10S)-5-(tert-butoxycarbonyl) 10-(R)-1-(tert-butyldimethylsilyl oxy)ethyl]-11-oxo-1,5 diazatricyclo[7.2.0.0(3,8)]undec-2- ene-2-carboxylate, was synthesized starting from an acetoxyazetidinone chiron 6 in a practical manner based on a C-C bond formation reaction between 6 and piperidinone-ester 5, palladium-catalyzed de(allyloxy)carbonylation of 7b and Wittig-type cyclization via an oxalimide 9. Selective deprotection of the N-Boc group of 2 was found to proceed smoothly by treatment with trimethylsilyl trifluoromethanesulfonate and 2,6-lutidine to give the amino compound 3, whose functionalization on the nitrogen atom to derivatives 10 followed by deprotection led to various 5-azatrinem acids 4. These compounds showed potent in vitro activities against gram-positive and gram-negative bacteria. PMID- 10823712 TI - Synthesis and alpha-adrenergic binding ligand affinities of 2-iminoimidazolidine derivatives. AB - In order to obtain possible veinotonic drugs acting through alpha2 receptor activation, we prepared clonidine analogues in which the 2-imino-imidazolidine was attached to various aliphatic or aromatic heterocycles. Among them, the two benzopyranic derivatives 16 and 22 exhibited interesting affinities (19 and 95 nM respectively on [3H]rauwolscine binding, compared to 35 nM for clonidine). Their affinity for alpha1 receptors was found to be much lower: 7570 and 5030 nM for 16 and 22 respectively, suggesting 16 to be 400 times more selective for alpha2 than for alpha1-adrenoceptors. PMID- 10823713 TI - Studies on bioadhesive granules I: granules formulated with Prosopis africana (prosopis) gum. AB - Prosopis gum (PG) extracted from Prosopis africana was investigated for bioadhesive delivery of theophylline (TPL). Bioadhesive granules containing TPL were formulated and the bioadhesive properties evaluated using adhesion of the granules onto a porcine intestinal mucus surface. The bioadhesion of the gum dispersion was also evaluated using coated glass beads and the strength of the films formulated from the gums was also determined. The release properties of the TPL-containing granules were assessed by diffusion of TPL from the granules through porcine intestinal wall into a sink solution. Sodium carboxymethylcellulose (SCMC) was used as the standard bioadhesive polymer. Results indicated that PG is highly bioadhesive compared to SCMC. The result of the release studies also showed that PG could be used to deliver TPL in a bioadhesive dosage form. PMID- 10823714 TI - Two new quinones from Iris bungei. AB - Two new benzoquinone derivatives, bungeiquinone (1) and dihydrobungeiquinone (2), and two known derivatives, 3-hydroxyirisquinone (3) and 3 hydroxydihydroirisquinone (4), were isolated from the roots of Iris bungei. The structures of the new compounds were established on the basis of spectroscopic methods. PMID- 10823715 TI - Synthesis and evaluation of some hydroxyproline-derived peptidomimetics as isoprenyltransferase inhibitors. AB - CA1A2X peptidomimetics containing a modified proline at position A2 were prepared and evaluated for their ability to inhibit farnesyltransferase (FTase) and geranylgeranyltransferase I (GGTase I) in enzymatic and cell-based assays. These compounds inhibited farnesylation of H-ras in vitro in the high nanomolar to low micromolar IC50 range. PMID- 10823716 TI - Delat9-tetrahydrocannabinol content in cannabis plants of greek origin. AB - The delta9-tetrahydrocannabinol (delta9-THC) content was identified and determined quantitatively using a Gas Chromatography Detector (Gas Chromatography Electron Ion Detector) instrument in samples of illicit herbal cannabis. Law enforcement authorities sent the samples to the Department of Forensic Medicine and Toxicology, University of Athens, for toxicological analysis. The concentrations of delta9-THC in these samples ranged from 0.08% to 4.41%. Such concentrations suggest that Greece might be at high risk, as an area for the illicit cultivation of "pedigree" cannabis plants. The forensic aspects of cannabis classification are discussed. PMID- 10823717 TI - Studies of the constituents of Gardenia species. II. Terpenoids from Gardeniae Fructus. AB - Four new terpenoids, gardenate A (1), 2-hydroxyethyl gardenamide A (2), (1R,7R,8S,10R)-7,8,11-trihydroxyguai-4-en-3-one 8-O-beta-D-glucopyranoside (3) and Jasminoside F (4), were isolated from Gardeniae Fructus. Their structures were established on the basis of spectral analysis. PMID- 10823718 TI - Sterol constituents from two edible mushrooms, Lentinula edodes and Tricholoma matsutake. AB - Two new sterols, (22E)-23-methylergosta-5,7,22-trien-beta-ol (1) and 5alpha,6alpha-epoxy-(22E)-ergosta-8,14,22-triene -3beta,7alpha-diol (2), have been isolated from two edible mushrooms, Lentinula edodes and Tricholoma matsutake, respectively, together with twelve known ones (3-14). The structures of the new compounds were elucidated on the basis of their spectral data. PMID- 10823719 TI - New megastigmane and tetraketide from the leaves of Euscaphis japonica. AB - New megastigmane (1) and tetraketide (2) were isolated from the leaves of Euscaphis japonica and the structures were elucidated by means of spectroscopic and chemical evidence. PMID- 10823720 TI - Studies on the 1,4-oxazepine ring formation reaction using the molecular orbital method. AB - 1,4-Oxazepine formation reactions of 1,8-naphthyridine derivatives (1-4) with peroxy acid have been studied using a semiempirical MO method (AM1) and an ab initio molecular orbital method (Gaussian 94). The energies of molecules involved in the reaction paths were calculated and the transition states related to experimental products were obtained. For the reactions of 1-3, the calculated energies of the transition states predicted the previously obtained products. However, the calculated values for the reaction of 4 suggested a different type of oxazepine compound, which was verified in further experiments. PMID- 10823721 TI - Serum parathyroid hormone and calcitonin levels in racehorses with fracture. AB - Serum parathyroid hormone (PTH) and calcitonin (CT) levels in fractured racehorses were measured by radioimmunoassay. Racehorses with fracture of large bone such as the radius, third metacarpus, third carpus, digital bone or tibia, showed normal PTH level and elevated CT level in the serum. Serum PTH level was slightly higher in racehorses with sesamoid bone fracture compared to that of healthy racehorses, but not statistically significant. Moreover, serum CT level of racehorses with sesamoid bone fracture was significantly higher than that of healthy racehorses. Racehorses with sesamoid bone fracture and large bone fracture might be in different conditions of calcium regulation. PMID- 10823722 TI - Apoptosis of proliferative cortical tubular epithelia in chronic progressive nephrosis of rats. AB - To investigate the role of apoptosis in the pathogenesis of rat chronic progressive nephrosis (CPN), the kidney of male F344/DuCrj rats, 19, 59, and 111 weeks of age, was examined histologically. In situ analysis for DNA fragmentation and proliferating cell nuclear antigen (PCNA) was performed simultaneously by TdT mediated dUTP-biotin nick end labeling (TUNEL) and immunohistochemistry, respectively. CPN was seen in all the kidneys of 59-week-old (n=6) and 111-week old rats (n=16), correlating significantly (p<0.01) with age. There were apoptotic bodies (ABs) in the single-layered epithelia of dilated tubules (SLD) and the multilayered epithelia (ML) of the cortical tubules. There were no ABs in any of the kidneys of the 19-week-old (n=5) or 59-week-old rats (n=6). Proliferative activity might have been enhanced in the single-layered and flattened epithelia, SLD, and ML of the cortical tubules in the kidneys of the 59 week-old rats (n=6) compared with that in 111-week-old rats (n=8). The correlations between the TUNEL-positive ratio and number of PCNA-positive cells, and age and the CPN grade were significant (p<0.01) exclusively in the ML. Thus, the results suggest that apoptosis occurs in the proliferative ML of rat CPN, and the pathological significance might be the removal of abnormal or excess cells. PMID- 10823723 TI - Multivariate analysis in skull osteometry of the common tree shrew from both sides of the Isthmus of Kra in Southern Thailand. AB - The Isthmus of Kra in Southern Thailand consists of a lowland of about 70 km in length. It has been suggested that the Isthmus may sink beneath the sea surface according to the change of level of the sea, and may function as a zoogeographical barrier in land mammals in this region. So, the geographical variation was osteometrically examined in skull of the common tree shrew (Tupaia glis) from the both sides of Isthmus of Kra. The osteometrical examination demonstrated that the skull is larger in southern population than in northern one. In the charts of the principal component analysis, however, the morphological separation between two populations can not be obviously seen in female. While, the results of the discriminant analysis indicated the morphological separation between the two populations. These findings suggest that the zoogeographical barrier of Isthmus of Kra may have influence on the osteometrical variation in the common tree shrew, when the Isthmus was covered with sea. PMID- 10823724 TI - Characteristic changes in carbohydrate profile in the kidneys of hereditary nephrotic mice (ICGN strain). AB - The ICR-derived glomerulonephritis (ICGN) mice consist of heterozygous and homozygous groups and are considered to be a good model for human idiopathic nephrotic syndrome. To reveal changes in cell-surface carbohydrate construction, 24 lectins were applied to kidney sections of 10-, 30- and 50-week-old male heterozygous and homozygous ICGN mice and age-matched male ICR mice. Bandeiraea simplicifolia lectin-I (BSL-I), which specifically binds to alpha-D galactopyranosyl groups, showed positive staining in the glomeruli of ICGN mice, but not in those of ICR mice. Positive BSL-I staining was observed only in distal tubules of homozygous ICGN mice. Lectin blotting for BSL-I demonstrated characteristic glycoproteins (45, 58 and 64 kD) in ICGN but not in ICR mice, and the levels of these molecules augmented in homozygous ICGN mice with the progression of renal failure. Moreover, succinylated wheat germ agglutinin, Dolichos biflorus agglutinin, Aleuria aurantia lectin and Ulex europaeus agglutinin-I showed positive staining only in the glomeruli of homozygous ICGN mice, but not in those of heterozygous ICGN or ICR mice. The staining intensities of Ricinus communis agglutinin-I, Phaseolus vulgaris agglutinin-E and -L, Lens culinaris agglutinin and Erythrina cristagalli agglutinin (ECL) in the glomeruli of homozygous ICGN mice were stronger than those of heterozygous ICGN and ICR mice. In conclusion, lectin histochemistry provided useful information for the diagnosis and prognosis of nephrotic lesions. Characteristic BSL-I binding glycoproteins may be pathogenic factors which cause renal disease in ICGN mice and are good tools to investigate the molecular mechanism of renal disorders in ICGN mice. PMID- 10823725 TI - Molecular characteristics and site specific distribution of the pigment of the silky fowl. AB - Silky fowl, a breed of chicken, is hyperpigmented in its various internal tissues. The pigment was extracted from various tissues of two strains of Silky fowl to determine its molecular structure and internal distribution. Analysis by infrared spectroscopy showed two spectrum patterns of the pigment in Silky fowl; one is from ovary and testis, the other is from periosteum and feather. The difference between the two spectra is possibly due to the sulfur contents of melanin. Especially, the spectra of the pigments from feather and periosteum shared the characteristics of synthesized melanin spectrum in common, which indicates that the melanocytes dispersed in these tissues were functionally the same. According to our quantitative analysis, the tissues examined were classified significantly in the order of the pigment content (p<0.05): periosteum > gonads (ovary or testis) = trachea > or = heart, liver, gizzard, cecum, muscles (Pectoralis and Supracoracoideus) and skin. In addition, the specific regions of embryonic neural crest derived cells, such as cardiac artery and various parts of cephalic tissues, were found to be locally hyperpigmented. These data suggest that hyperpigmentation (fibromelanosis) in Silky fowl chicken occurs in a tissue- and organ-specific manner, which is strongly related to neural crest cell development. It is hypothesized that neural crest cells of the bird, containing melanocyte progenitors, acquire unusual ability to differentiate into melanocytes excessively, and to extend the distribution of their descendant along the destinations of neural crest derivatives. PMID- 10823726 TI - Kinetics of lymphocytic subsets in chicken tracheal lesions infected with infectious bronchitis virus. AB - The kinetics of T-cells (CD3 positive (+), CD4+ and CD8+ cells) and B-cells (IgG+, IgM+ and IgA+ cells) in chicken trachea were studied immunohistochemically and histopathologically following an intratracheal inoculation of infectious bronchitis virus (IBV). Viral antigen was detected in the cytoplasm of tracheal epithelium from 16 hr to 6 days post-inoculation (p.i.) with a peak on 4 days p.i. A few IgG+, IgM+ and IgA+ cells were detected in the submucosa from 8 hr p.i. Thereafter IgG+ and IgM+ cells were gradually increased in number, and dramatically increased from 3 days p.i., peaked on 4 days p.i., and gradually decreased after 5 days p.i. IgA+ cells were detected in a small number than IgG+ and IgM+ cells during the all experimental period. These B cells mainly existed in the lamina propria, and some cells were recognized in the interepithelial space. After 14 days p.i., small number of IgG+ and IgM+ cells were detected in the germinal center of lymph follicles in the lamina propria. From 24 to 60 hr p.i., a few number of CD3+, CD4+ and CD8+ cells were detected at the perivascular area in the lamina propria. After 3 or 4 days p.i., each positive T-cells increased rapidly in number, and reached on the peak at 5 days p.i. CD3+, CD4+ and CD8+ cells tend to distribute diffusely, perivascular area, and surrounding area of CD4+ cells, respectively. CD4+ cells were dramatically decreased from 7 days p.i., and CD3+ and CD8+ cells were decreased from 14 days p.i. No T-cells were detected in the lymph follicles in the lamina propria. PMID- 10823727 TI - Effects of oral administration of "rumen-bypass" vitamin D3 on vitamin D and calcium metabolism in periparturient cows. AB - Eleven late-pregnant Jersey cows were assigned to two groups; a group (PO-RBVD group) consisting of five cows treated with an oral administration of 10 million I.U. of an encapsulated form of vitamin D3 ("rumen-bypass" VD3; RBVD3) and another group (IMVD group) consisting of the other six treated with an intramuscular injection of 10 million I.U. of vitamin D3 (VD3). The cows received the RBVD3 or VD3 administration at 7 days before the expected parturition. The changes in the plasma concentrations of vitamin D metabolites, ionized Ca (Ca++) and inorganic phosphorus (iP) were evaluated. Of the vitamin D metabolites, the plasma 25-hydroxyvitamin D concentrations in PO-RBVD group increased significantly after the RBVD3 administration and remained in high levels that were significantly higher than those in IMVD group. This suggested that RBVD3 was absorbed rapidly and excellently from the post-ruminal digestive tract without the degradation by ruminal microorganisms. The plasma Ca++ and iP concentrations in PO-RBVD group tended to be higher after the administration and around parturition than those in IMVD group. From these observations, it was suggested the oral RBVD3 administration had more potent ability to prevent parturient paresis compared with the VD3 injection used widely in Japan. PMID- 10823728 TI - Sympathetic activation of hepatic and splenic IL-1beta mRNA expression during oscillation stress in the rat. AB - Interleukin (IL)-1beta mRNA expression in the liver and spleen was examined after subjection to oscillation stress in the rat. Thirty-minute subjection to oscillation stress increased IL-1beta mRNA expression in the both organs. Prior treatment of rats with gadolinium chloride, which eliminates macrophages, prevented the stress-induced IL-1beta expression. Either adrenalectomy or treatment of guanethidine, a blocker of norepinephrine release in the sympathetic nerve endings, partially attenuated the stress-induced response, but the combined treatment completely blocked it. Injection of beta-adrenergic antagonist (propranolol) also suppressed the stress-induced response. These results suggest that oscillation stress induces IL-1beta mRNA expression in the liver and spleen, probably in Kupffer cells and splenic macrophages, and that stress-induced IL 1beta expression is elicited by catecholamines released from sympathetic nerve terminals and the adrenal gland. PMID- 10823729 TI - Serum progesterone and estradiol-17beta concentrations in captive and free ranging adult female japanese black bears (Ursus thibetanus japonicus). AB - Progesterone (P4) and estradiol-17beta (E2) concentrations were measured in serum samples obtained from 23 captive and 23 free-ranging adult female Japanese black bears. We then determined the relationship between changes in these sex steroid hormones and pregnancy. In all captive bears, which included animals of both known and unknown reproductive status, serum P4 concentrations were low from April to July, then tended to become higher after August. The levels then became much higher still in November and December, but returned to low levels in March. Serum P4 concentrations in eight captive pregnant bears, which had parturitions the following spring, increased gradually from August (0.5-2.4 ng/ml) to October (0.9-3.6 ng/ml), and achieved significantly higher maximum levels in December (7.2-18.0 ng/ml). Thereafter, serum P4 concentrations tended to decrease (3.5-6.4 ng/ml in January and 0.3-0.7 ng/ml in March). In all captive bears, serum E2 concentrations varied from April to October but showed low levels in November and December, and became high in January. Serum E2 concentrations in the eight pregnant bears were high in May (95.6-191.4 pg/ml) and varied from August to October (35.6-143.3 pg/ml). Subsequently, serum E2 concentrations in December dropped to significantly lower minimum levels (5.3-11.9 pg/ml) and increased again in January (67.6-153.1 pg/ml). Among the free-ranging bears, the data on serum P4 concentrations in eight bears led to expectations of pregnancy, whereas serum E2 concentrations showed no distinct evidence related to pregnancy. These results, particularly in captive pregnant bears, indicate that a marked increase of P4 in December might be accompanied by reactivation of the corpus luteum preceding implantation. Furthermore, changes in E2 concentrations suggested the possibility that a decline in December and an increase in January are associated with implantation and parturition, respectively. PMID- 10823730 TI - The risk of contrast media-induced ventricular fibrillation is low in canine coronary arteriography with ioxilan. AB - Previous studies have proposed that sodium supplement to nonionic contrast media (CM) can decrease the risk of ventricular fibrillation (VF). This study was designed to compare the occurence of VF induced by ioxilan (containing 9 mmol/LNa+) with other nonionic CMs. After wedging a catheter in the right coronary artery, test solutions including ioxilan, ioversol, iomeprol, and iopromide were infused for 30 sec at the rate of 0.4 ml/sec or until VF occurred. Then, incidence of VF, contact time (i.e. the time required to produce VF), and QTc were measured. Also, the CMs other than ioxilan were investigated at sodium levels adjusted to 9 and 20 mmol/L Na+. The incidence of VF with ioxilan (0%) was the lowest of all. In the other CMs, the incidence decreased in accordance with increase of sodium. Iomeprol and iopromide showed significant reduction of VF incidence at the sodium level of 20 mmol/L. The higher sodium supplements also prolonged the contact times. The increase of QTc was the greatest in ioxilan. Ioxilan has the least arrythmogenic property among the current low-osmolality nonionic CMs. This property might be attributable to an optimal sodium concentration of 9 mmol/L in the CM. PMID- 10823731 TI - Establishment and characterization of the MSKR inbred strain originated from Japanese wild mice (Mus musculus molossinus). AB - A new inbred strain, MSKR, originated from Japanese wild mice was established in April, 1998. The MSKR mice were 60% of the C57BL/6N inbred mice in the 60-day body weight. Tail length/head-body length and hind-foot length/head-body length of the MSKR mice were significantly smaller than those of the C57BL/6N mice (0.896 vs 1.061, 0.189 vs 0.204), but ear length/head-body length of the MSKR mice was significantly larger than that of the C57BL/6N mice (0.143 vs 0.137). The age of the first parturition and size of the first litter were 63.20 +/- 2.71 days and 6.20 +/- 0.37, respectively, at the 20th and 22nd inbreeding generations. Genetic characterization of the MSKR strain was performed using 34 microsatellite markers, 29 biochemical markers, 9 immunogenetic markers, 3 coat color markers, and mitochondrial DNA RFLP-haplotypes. The result indicated that this newly established inbred strain has some different gene constitution from already known molossinus and common laboratory strains. PMID- 10823732 TI - Dose-related cardiovascular effects of isoflurane in chickens during controlled ventilation. AB - We studied cardiovascular effects of isoflurane in chickens during controlled ventilation. Following the determination of the minimal anesthetic concentration from the response to clamping of a digit, dose-related effect of isoflurane on heart rate and arterial pressure were studied. The minimal anesthetic concentration of isoflurane was 1.25 +/- 0.13% (mean +/- SD, n=9). There was a dose-dependent decrease in arterial pressure. The heart rate did not change significantly over a range of 1 to 2 times the minimal anesthetic concentration. PMID- 10823733 TI - An epizootiological survey of necropsy cases (1993-1997) at University of the Philippines. AB - An epizootiological survey of necropsied cases (1993-1997) at University of the Philippines was performed. A total of 368 cases included 238 avian and 111 porcine cases. Amongst avian cases, the major cause of death was infectious diseases in 212 (89%) cases including 97 (41%) bacterial, 36 (15%) viral, and 21(9%) parasitic diseases. The majority of the avian bacterial diseases presented as septicemia (73 cases) and the viral diseases as Newcastle disease (17 cases). In porcine cases, the major cause of death was also infectious diseases, in 100 (90%) cases including 52 bacterial and 29 viral diseases. Porcine bacterial diseases were classified into 36 septicemia, 4 hemophillosis and 4 colibacillosis. Amongst the porcine viral diseases, most cases were diagnosed as Hog cholera (22 cases). PMID- 10823734 TI - Spontaneous activities measured continuously by an accelerometer in beagle dogs housed in a cage. AB - Spontaneous activities in 10 beagle dogs housed in an individual cage were recorded continuously for 2 hr by an accelerometer fixed to dogs and by a video camera. The responses of the accelerometer were compared to movements identified from the video tapes. We found that gross differentiation of quantitative spontaneous activities might be possible by using only the accelerometer if the threshold of the accelerometer and the amount of acceleration volume were set adequately: the responses of the accelerometer at the threshold of 0.10 G with the acceleration number of 251 or more revealed only movements of the whole-body, and those at the threshold of 0.02 G all movements including changes of the part(s) of the body. PMID- 10823735 TI - A histochemical study of the camel (Camelus bactrianus) duodenal glands. AB - The complex carbohydrates in the camel duodenal glands were examined histochemically at light and electron microscopic levels. The duodenal glands of the camel were distributed in the submucosa 2 m caudal from the pylorus. These were branched tubuloalveolar glands. The terminal portion of each lobule was formed by only one type of mucous cell. The duodenal gland cells contained acidic and neutral carbohydrates. The mucous cells mainly contained sulfate and carboxyl carbohydrate with sialic acid, and they also contained a few neutral carbohydrates with different saccharide residues such as mannose, glucose, galactose, N-acetyl glucosamine and N-acetyl galactosamine. The results showed that the secretary granules of the duodenal glands in the camel contain mainly acidic carbohydrates. These findings seem to be the morphological characteristics of the duodenal glands in the camel. PMID- 10823736 TI - Plasma renin activities, angiotensin II concentrations, atrial natriuretic peptide concentrations and cardiopulmonary function values in dogs with severe heartworm disease. AB - Relationships among plasma renin activities (PRA), plasma angiotensin II (ATII) concentrations, atrial natriuretic peptide (ANP) concentrations and cardiopulmonary function values were examined in dogs with ascitic pulmonary heartworm disease and acute- and chronic-vena caval syndrome (CS). PRA, plasma ATII concentration and plasma ANP concentration tended to be higher or were significantly higher in dogs with ascites, acute- and chronic-CS. PRA correlated significantly with plasma ATII concentration, WBC count, ALP activity, plasma concentrations of urea nitrogen, creatinine, sodium, potassium, and chloride, right ventricular endodiastolic pressure and right atrial pressure. Plasma ATII concentration correlated significantly with WBC count, plasma concentrations of urea nitrogen, sodium, and potassium, right ventricular endodiastolic pressure and right atrial pressure. Plasma ANP concentration did not correlate with PRA or ATII concentration, but correlated significantly only with pulmonary arterial pressure. PMID- 10823737 TI - Active-oxygen involvement in canine NK-mediated cytotoxicity. AB - We examined the relationship between natural killer (NK) cell-mediated cytotoxicity, the produced active-oxygen and cytotoxic factor (CF) release in co culturing canine NK cells with tumor cells (CL-1 target cells). In co-culturing, the adding of n-propyl gallate (active-oxide scavenger) removed the produced active-oxygen, which inhibited NK cell-mediated cytotoxicity and the CF release. Moreover, adding of this agent inhibited the tyrosine phosphorylation of NK intracellular protein which observed in co-culturing. Therefore, the active oxygen produced from canine NK cells are thought to relate the signal transduction in NK-mediated cytotoxicity. PMID- 10823738 TI - Myeloblastic leukemia with massive neoplastic infiltration of the skin and mediastinum in a cow. AB - An acute myeloblastic leukemia was found in a 3.5-year-old Holstein cow. The neoplasm was characterized by massive tumor growths, and there were multiple tumor nodules in the dermis or subcutis and a large tumor mass in the mediastinum. This tumor showed negative reactivity for CD3, CD79a, major histocompatibility complex class II and myeloid/histiocyte antigen. Ultrastructural features such as dispersed cytoplasmic granules and poorly developed organelles were compatible with those of early promyelocytes. PMID- 10823739 TI - Effect of linoleic acid-albumin on post-thaw survival of in vitro-produced bovine embryos at the 16-cell stage. AB - The effect of addition of linoleic acid-albumin (LAA) to culture medium before freezing on the survival rate of bovine 16-cell embryos after freezing-thawing was investigated. Embryos were incubated in CR1aa containing LAA (0.25 mg/ml) for 4 days after insemination. A conventional slow cooling method was used, in which embryos were cooled at a rate of 0.3 degrees C/min to -30 degrees C in medium supplemented with 1.5 M ethylene glycol and 0.2 M trehalose. The developmental rate to the blastocyst stage of thawed embryos that had been cultured with LAA containing medium before freezing was higher than that of these cultured without LAA (P<0.05). However, with fresh, non-frozen, embryos that were incubated under the same culture conditions (with and without LAA), no such difference was found. PMID- 10823740 TI - Retinoid receptors and the induction of apoptosis in canine osteosarcoma cells. AB - Retinoids, all-trans-retinoic acid (ATRA) and 9-cis-retinoic acid (9-cis-RA), induced morphological changes and apoptosis-like cell death characterized by cell shrinkage, chromatin condensation and nuclear disintegration in three canine osteosarcoma cells, OOS, HOS and POS, at a concentration of 10(-5) M. Both retinoid receptors, RARs and RXRs, were identified in these cells. 9-cis-RA bound to both the RXRs and the RARs, whereas ATRA bound to only the RARs in these cells. Those results indicate that the induction of apoptosis in canine osteosarcoma cells may be mediated by the specific control of RARs and RXRs. PMID- 10823741 TI - Modernising Social Services. PMID- 10823742 TI - Relationships between the development of biological risk factors for coronary heart disease and lifestyle parameters during adolescence: The Northern Ireland Young Hearts Project. AB - The purpose of this study was to examine relationships between the longitudinal development of biological risk factors for coronary heart disease (CHD) (namely, serum lipids, body fatness, blood pressure and cardiorespiratory fitness) and the development of lifestyles considered to be atherogenic. The study was conducted in a representative sample of adolescents from Northern Ireland, a region of high coronary mortality, and atherogenic aspects of lifestyle such as cigarette smoking, physical inactivity and selected aspects of diet were examined. Repeated measurements were made at 12 and 15 y of age on 229 boys and 230 girls. Longitudinal relations were analysed with generalised estimating equations, and the following longitudinal relations were found: for boys diastolic blood pressure was positively associated with vitamin C intake (P = 0.014), and inversely with energy intake (P = 0.006), and smoking (P = 0.048). Systolic blood pressure was inversely related to physical activity (P = 0.012), and smoking (P = 0.000). Body fatness was also inversely related to smoking (P = 0.006). Total cholesterol (TC) was positively related to physical activity (P = 0.044) and the TC:HDL cholesterol ratio positively to vitamin C intake (P = 0.008). Cardiorespiratory fitness was positively related to physical activity (P = 0.000) and inversely to smoking (P = 0.031). For girls, systolic blood pressure was positively related to vitamin C intake (P = 0.042); HDL cholesterol inversely to carbohydrate intake (P = 0.014), fat intake (P = 0.031), cholesterol intake (P = 0.042) and smoking (P = 0.035) and positively to energy intake (P = 0.035). The TC:HDL cholesterol ratio was inversely related to energy intake (P = 0.038) and finally, cardiorespiratory fitness positively to physical activity (P = 0.001). These results offer additional evidence that changes in lifestyle are associated with changes in biological risk factors in adolescents. Education and intervention at this stage, particularly in relation to cigarette smoking, physical activity and certain aspects of diet appear justified as part of a preventative strategy for CHD. PMID- 10823743 TI - An audit on access to coronary artery surgery within a health district using New Zealand priority criteria as a benchmark. AB - A professional advisory group in New Zealand, consisting of cardiologists, cardiac surgeons and general practitioners have recommended priority criteria for access to Coronary-Artery Bypass Graft (CABG). They used published outcome studies in developing the criteria, and numerical scores were assigned to both biological and social dimensions of ischaemic heart disease. The New Zealand criteria were used to investigate access to services for patients in the UK in Sunderland, Tyne and Wear. Case notes of 229 Sunderland residents who underwent coronary angiography in the 1995 96 financial year were reviewed. The clinical threshold for access to CABG was much higher in Sunderland than in New Zealand. After controlling for the severity of disease, there was no evidence of inequity of access based on age, gender, smoking status or the deprivation indices of the patients' ward of residence. The correlation between the New Zealand Priority Criteria scores and the waiting time for coronary angiography, CABG and Percutaneous Transluminal Coronary Angioplasty (PTCA) was present, but weak (r = 0.22, -0.1 and -0.54 respectively). The New Zealand consensus criteria proved a useful benchmark both to assess access for patients with ischaemic heart disease to coronary artery surgery in a local Health District and to prioritise patients on the waiting list for coronary artery surgery. They allowed a realistic assessment of access according to age, gender, smoking status and deprivation by taking disease severity into account. PMID- 10823744 TI - Health education pamphlets about smoking--their benefit to smokers and non smokers. AB - The aim of this present study was to compare the use by smokers and non-smokers of pamphlets about smoking as delivered from different settings. The study was a nation-wide cross-sectional survey of 1924 randomly selected, Danish men and women, aged 14-77 y, who had answered a mailed questionnaire in 1994. Of these 71% also participated in a telephone interview enquiring about the use of health education material, smoking status and socio-demographic variables, 39% of readers of household-delivered anti-smoking pamphlets reported having gained information from them and 22% reported having made changes in their own smoking behaviour such as avoiding smoking in the presence of non-smokers. In general practice settings, these percentages were higher among smokers. Smokers who were thinking of stopping smoking in the near future were in addition more likely to take and to read smoking related health education materials from other places. Non-smokers received (3 49%) and read pamphlets about smoking as frequently as did smokers who did not intend to quit. In conclusion, written health education material was well received by readers, but, when distributed in a more open setting it needs to be targeted towards smokers who are considering stopping smoking. In general practice, smokers not thinking of stopping were open to health education, and pamphlets used in this setting should also target this group. Non-smokers contribute indirectly to smokers quitting by providing support to smokers and pamphlets for non-smokers need to be more targeted towards this social role. PMID- 10823745 TI - Perinatal mortality in Taiwan. AB - Information on perinatal deaths was obtained from 310 women by collecting detailed obstetric histories dating from marriage to the start of the survey. These histories were compared to those of 688 age matched controls. Potential risk factors, levels and time trends of perinatal mortality in Taiwan were examined and factors underlying stillbirths and early neonatal deaths were also compared using conditional logistic regression analyses. A nearly 56% decline of the perinatal mortality rate during the 35 y, approximately, prior to the survey was observed. Risk of stillbirths was increased among those who had abused illegal drugs during pregnancy, those who reported that the pregnancy was unwanted and those with Thalassemia trait. Body mass index was log-linearly related with stillbirths, with higher body mass associated with higher risk. For early neonatal deaths, those mothers aged 19 y or less, those giving birth to either their first children or to their fifth or later child, those who had their first prenatal care visit after the first three months of pregnancy were associated with increased risk in the logistic model. Those with a birth interval of less than two years and those with less education were associated with increased risk in both perinatal death groups. While some of these factors have already been associated with perinatal deaths, others have not; the new associations provide clues to mechanisms by which the risk of death increases before or after delivery. PMID- 10823746 TI - Knowledge, attitudes, beliefs and practices about HIV/AIDS among the overseas job seekers in Bangladesh. AB - A study of the knowledge, attitudes, beliefs and practices (KABP) relating to HIV/AIDS was conducted among people from Bangladesh seeking work overseas (N = 300), during February, 1997 and March, 1997. Only 26% of the respondents knew of AIDS and out of 13 basic facts concerning HIV/AIDS the mean score of the sample was 1.63 correct responses. Most of those who knew of HIV had some false beliefs about the mode of HIV transmission, for example, believing that HIV could be contracted by touching an AIDS patient, or sharing bathing facilities or eating utensils. Sex with brothel-based commercial sex workers (100%), sharing contaminated needles (93.6%) and blood transfusion from infected individuals (93.6%) were seen as the main route of HIV transmission. Printed media (69%) was the main source of AIDS information. Multiple logistic regression analysis showed that having a non-agricultural occupation (P < 0.04), being resident in Dhaka, Chandpur, Noakhali, Comilla, and Chittagong (P < 0.01), being in the habit of reading newspapers (P < 0.05), using condoms (P < 0.04), having heard about condoms (P < 0.003), having seen condoms (P < 0.005) and knowing where to buy condoms (P < 0.0005) were significantly associated with AIDS awareness. There is insufficient AIDS awareness among overseas job seekers which calls for public initiatives to provide AIDS information to them before they leave Bangladesh to work abroad. PMID- 10823747 TI - Sub-national response in HIV/AIDS: a case study in AIDS prevention and control from Sindh province, Pakistan. AB - HIV/AIDS in Pakistan is slowly gaining recognition as a public health issue of great importance. However, the responses to the disease have been marred by lack of coordination and commitment. We examine, in this paper, the situation in the Sindh province of Pakistan, which is recognized as having the sole fully functioning AIDS prevention and control programme in the country. In discussing the results of the Sindh programme's activities we highlight progress made as well as gaps in data and surveillance. We also recommend strategies for implementation at the provincial and national levels. In addition this example of a sub-national government programme provides a case study for similar programmes in the region. PMID- 10823748 TI - Summary of lecture given to the Faculty of Community Health, Annual General Meeting on 20th October 1998 by Patricia M Sonksen. Severely visually impaired children: neurodevelopmental concerns. PMID- 10823749 TI - A relationship between the evolution of hepatitis C virus variants, liver damage, and hepatocellular carcinoma in patients with hepatitis C viremia. AB - To clarify the mechanism of liver damage induced by hepatitis C virus (HCV) and to determine whether the damage is related to hepatocellular carcinoma (HCC), HCV RNA levels were measured serially, and HCV genome mutations were analyzed from serum of 274 Japanese patients with chronic HCV viremia during 1993-1998. All patients had alanine aminotransferase (ALT) levels measured during 1986-1998. Patients with consistently normal ALT levels had identical and highly conserved HCV core regions; however, those with consistently abnormal ALT levels had quasi species, and the population of the quasi species changed over time. HCV RNA levels did not change in the 274 patients. HCC developed in 31% of 80 patients with consistently abnormal ALT levels and in 4% of 92 patients with intermittently abnormal ALT levels but never in 102 patients with ALT levels consistently normal during 1993-1998. In patients with chronic HCV viremia, persistent liver damage plays an important role in the development of HCC. PMID- 10823750 TI - Association of hepatitis C virus-specific CD8+ T cells with viral clearance in acute hepatitis C. AB - CD8+ T lymphocytes play a major role in antiviral immune defense. Their significance for acute hepatitis C is unclear. Our aim was to correlate the CD8+ T cell response with the outcome of infection. Eighteen patients with acute hepatitis C and 19 normal donors were studied. Hepatitis C virus (HCV)-specific CD8+ T cells were identified in the enzyme-linked immunospot assay by their interferon-gamma (IFN-gamma) production after specific stimulation. The highest numbers of IFN-gamma-producing HCV-specific CD8+ T cells were found in patients with acute hepatitis C and a self-limited course of disease during the first 6 months after onset of disease, but these numbers dropped thereafter to undetectable levels. The differences in responsiveness between patients with self limited disease versus patients with a chronic course were statistically significant (P<.001). Our data show that the number of IFN-gamma-producing HCV specific CD8+ T cells during the first 6 months after onset of disease is associated with eradication of the HCV infection. PMID- 10823751 TI - Cytotoxic T lymphocyte (CTL) responses to human cytomegalovirus pp65, IE1-Exon4, gB, pp150, and pp28 in healthy individuals: reevaluation of prevalence of IE1 specific CTLs. AB - The prevalence of human cytomegalovirus (HCMV) pp65-, pp150-, IE1-exon4-, gB- and pp28-specific cytotoxic T lymphocyte (CTL) responses was compared among 34 healthy individuals, grouped by neutralizing antibody titers. Moderately and highly seropositive donors showed predominantly pp65- and IE1-exon4-specific CTL responses (92% and 76% of the donors, respectively), with similar precursor frequencies in the 2 donors tested. In addition, highly seropositive and a few moderately seropositive donors showed CTL responses to gB and pp150 (33% and 30% of the donors, respectively). No individual recognized pp28 as a target in the CTL assay. Phenotypic analysis revealed a mixed effector population of CD4+ and CD8+ (1 donor) or only CD8+ cells for pp65-specific effectors (2 donors). IE1 exon4- and pp150-specific effectors were CD8+ (2 donors and 1 donor, respectively), whereas gB-specific CTLs were CD4+ (1 donor). These data may help to design a cellular immunity-based vaccine effective against HCMV diseases. PMID- 10823752 TI - Genetic diversity and molecular epidemiology of the G protein of subgroups A and B of respiratory syncytial viruses isolated over 9 consecutive epidemics in Korea. AB - To study genetic variation and molecular epidemiology of the G protein of respiratory syncytial virus (RSV), 253 strains from a children's hospital in Korea over 9 consecutive epidemics were analyzed. Restriction analysis of the entire G protein gene demonstrated 24 genotypes among 188 subgroup A and 6 among 65 subgroup B isolates. Two to 4 dominant genotypes of subgroup A cocirculated, and different genotypes predominated in each epidemic. Predominant genotypes were replaced with new genotypes during consecutive epidemics. One of 2 dominant genotypes among subgroup B predominated alternately or concurrently. Phylogenetic analysis revealed that there were multiple lineages, with clustering related to their location and time of isolation among strains from Korea and worldwide. Geographic and temporal distinction have been shown more clearly for subgroup B than subgroup A. These results suggest that the G protein of RSV is continuously evolving, with a distinct pattern presumably due to immune selection in a localized region over time. PMID- 10823753 TI - The prevalence of human herpesvirus-7 in renal transplant recipients is unaffected by oral or intravenous ganciclovir. AB - The purpose of this study was to compare the prevalence of human herpesvirus (HHV)-7 and cytomegalovirus (CMV) viremia and the effects of oral and intravenous (iv) ganciclovir in renal transplant recipients at risk for CMV. Stored lysates from peripheral blood leukocytes from 92 patients, who had been previously analyzed for CMV viremia by polymerase chain reaction (PCR) for 12 weeks after transplantation, were analyzed for HHV-7 viremia. Baseline and peak prevalences of HHV-7 viremia were 22% and 54%, respectively (P<. 0001). Eighty-two (89%) of 92 patients had at least 1 positive PCR for HHV-7. Oral ganciclovir and treatment with iv ganciclovir had no effect on the prevalence of HHV-7 viremia. In contrast, CMV was almost completely suppressed in patients who received oral ganciclovir, and when present, CMV responded to iv therapy. These results indicate that HHV-7 is resistant to ganciclovir at levels that were effective for prevention and treatment of CMV. PMID- 10823754 TI - Human herpesvirus 8 in Brazilian Amerindians: a hyperendemic population with a new subtype. AB - Human herpesvirus 8 (HHV-8) epidemiology in Brazilian Amerindians was studied. Use of an immunofluorescence (IFA) test for latent antibody demonstrated that the prevalence of HHV-8 in 781 Amerindians of diverse tribes (overall, 53% prevalence) was not related to language group or sex but rather increased gradually from 41% in children <10 years of age to 65% in adults >/=30 years of age. In IFA-positive subjects, HHV-8 DNA was detected in 3 (16%) of 19 mononuclear cell samples from peripheral blood and in 1 of 16 saliva samples. The sequences of conserved ORF22 and K6 genes were typical of HHV-8, but the variable K1 gene sequences were only 70%-75% identical to other known HHV-8 strains. Thus, a new HHV-8 subtype, E, is hyperendemic in Brazilian Amerindians, although Kaposi's sarcoma has not been reported. Transmission is probably oral rather than sexual. The limited genetic pool in isolated groups may permit more frequent transmission of a virus with a low prevalence in heterogeneous populations. PMID- 10823755 TI - Outbreak of hantavirus infection in the Four Corners region of the United States in the wake of the 1997-1998 El Nino-southern oscillation. AB - Hantavirus cardiopulmonary syndrome (HCPS), a rodent-borne zoonosis, has been endemic in the Americas for at least several decades. It is hypothesized that the 1991-1992 El Nino-southern oscillation (ENSO) caused increased precipitation that allowed an increase in rodent population densities, thereby increasing the possibility of transmission to humans. The result was a 1993-1994 outbreak of the disease in the Four Corners states of the southwestern United States. A second strong ENSO occurred in 1997-1998, after a period of considerable public education about the risks of hantavirus infection that began during the 1993-1994 outbreak. The caseload of HCPS increased 5-fold above baseline in the Four Corners states in 1998-1999. Regions that had received increased rainfall in 1998 were especially affected. A large majority of the 1998-1999 case patients reported indoor exposure to deer mice. Hantavirus outbreaks can occur in response to abiotic events, even in the face of extensive public education and awareness. PMID- 10823756 TI - Induction of human immunodeficiency virus type 1 expression by anaerobes associated with bacterial vaginosis. AB - Bacterial vaginosis (BV) is a common disorder characterized by increased levels of anaerobic bacteria in the genital tract. BV has been associated with an increased rate of sexual transmission of human immunodeficiency virus (HIV). The effects of BV-associated anaerobic bacteria on HIV expression in monocytoid cells and T cells were examined. Peptostreptococcus asaccharolyticus and Prevotella bivia stimulated HIV expression in monocytoid cells, whereas Bacteroides ureolyticus, Peptostreptococcus anaerobius, and Lactobacillus acidophilus did not enhance HIV expression. P. asaccharolyticus also enhanced HIV expression in T cells and activated HIV long-terminal-repeat transcription in U38 cells. This report suggests a mechanism by which disturbances in vaginal flora could lead to a higher rate of sexual transmission of HIV. Furthermore, this study supports the idea that treatment of BV might serve as a preventive measure to reduce the risk of HIV transmission. PMID- 10823757 TI - Influence of HLA supertypes on susceptibility and resistance to human immunodeficiency virus type 1 infection. AB - Certain human leukocyte antigens, by presenting conserved immunogenic epitopes for T cell recognition, may, in part, account for the observed differences in human immunodeficiency virus type 1 (HIV-1) susceptibility. To determine whether HLA polymorphism influences HIV-1 susceptibility, a longitudinal cohort of highly HIV-1-exposed female sex workers based in Nairobi, Kenya, was prospectively analyzed. Decreased HIV-1 infection risk was strongly associated with possession of a cluster of closely related HLA alleles (A2/6802 supertype; incidence rate ratio [IRR], 0.45; 95% confidence interval [CI], 0.27-0.72; P=.0003). The alleles in this supertype are known in some cases to present the same peptide epitopes for T cell recognition. In addition, resistance to HIV-1 infection was independently associated with HLA DRB1*01 (IRR, 0.22; 95% CI, 0.06-0.60; P=.0003), which suggests that anti-HIV-1 class II restricted CD4 effector mechanisms may play an important role in protecting against viral challenge. These data provide further evidence that resistance to HIV-1 infection in this cohort of sex workers is immunologically mediated. PMID- 10823758 TI - Influence of human immunodeficiency virus-infected maternal environment on development of infant interleukin-12 production. AB - Monocyte-derived cytokine production by cord blood mononuclear cells (CBMC) from infants born to human immunodeficiency virus (HIV)-positive and -negative women was measured to determine whether monocyte dysfunction could contribute to the accelerated HIV disease of pediatric patients. Production of interleukin (IL)-12, but not that of tumor necrosis factor-alpha and IL-10, was reduced, compared with adult peripheral blood mononuclear cells (PBMC). This deficiency was more pronounced in infants of HIV-positive women, whose IL-12 production was also deficient. CBMC IL-12 levels were increased by interferon-gamma and CD40 ligand but remained deficient, compared with PBMC. IL-12 production was undetectable in 7 of 8 HIV-positive infants, in contrast to 21 of 26 uninfected infants. Uninfected infants of infected women exhibited an intermediate profile. These findings suggest that the maternal environment and/or exposure in utero to HIV products influence the newborn's immune response and that the differences between infants born to HIV-positive and -negative women may persist. PMID- 10823759 TI - Disease progression and survival with human immunodeficiency virus type 1 subtype E infection among female sex workers in Thailand. AB - This study describes rates and correlates of disease progression and survival among 194 female sex workers in northern Thailand who were infected with human immunodeficiency virus type 1 (HIV-1; 96% with subtype E). The median rate of CD4 T lymphocyte decline (3.9 cells/microL/month), median time from infection to <200 CD4 T lymphocytes/microL (6.9 years), and time to 25% mortality (6.0 years) were similar to those found in studies performed in Western countries before highly active antiretroviral therapy was available to populations infected with HIV-1 subtype B. Mortality rates among women with >100,000 HIV-1 RNA copies/mL were 15.4 times higher (95% confidence interval, 5.2-45.2) than among women with <10,000 copies. Initial CD4 T lymphocyte counts and serum virus load were independently strong predictors of survival. These results can help in assessing the effects of the epidemic in Thailand and in determining the prognoses for individual patients. PMID- 10823760 TI - Neutralization of human immunodeficiency virus type 1 (HIV-1) mediated by parotid IgA of HIV-1-infected patients. AB - Infection with human immunodeficiency virus type 1 (HIV-1) has been shown to elicit a serum antibody response with neutralizing activity against T cell line adapted HIV strains and primary HIV-1 isolates. Mucosal surfaces are the primary route of HIV-1 infection. Evidence is presented here for the presence of HIV neutralizing antibodies in secretions. Infection of mucosal cells with HIV stimulates systemic and mucosal immune responses and results in the generation of neutralizing antibodies. Serum IgG and IgA neutralize HIV-1MN infection of susceptible T cell lines; serum IgG inhibits more effectively. Mucosal IgA purified from parotid saliva of HIV-1-seropositive individuals could neutralize both a T cell line-adapted strain and a primary isolate. The neutralizing activity of IgA was not directed against the anti-third-variable-loop or the anti ELDKWA epitope. Thus, the specificity of mucosal IgA for HIV-1 neutralization epitopes remains to be determined and may provide insight into development of a mucosal vaccine. PMID- 10823761 TI - A randomized, controlled, phase II trial comparing escalating doses of subcutaneous interleukin-2 plus antiretrovirals versus antiretrovirals alone in human immunodeficiency virus-infected patients with CD4+ cell counts >/=350/mm3. AB - A total of 73 patients with baseline CD4+ cell counts >/=350 cells/mm3 who were receiving combination antiretroviral therapy (ART) were randomized to receive subcutaneous interleukin-2 (IL-2; n=36) in addition to ART or to continue ART alone (n=37). Subcutaneous IL-2 was delivered at 1 of 3 doses (1.5 million international units ?MIU, 4.5 MIU, and 7.5 MIU per dose) by twice-daily injection for 5 consecutive days every 8 weeks. After 24 weeks, the time-weighted mean change from baseline CD4+ cell count was 210 cells/mm3 for recipients of subcutaneous IL-2, compared with 29 cells/mm3 for recipients of ART alone (P<.001). There were no significant differences between treatment groups for measures of plasma human immunodeficiency virus RNA (P=.851). Subcutaneous IL-2 delivered at doses of 4.5 MIU and 7.5 MIU resulted in significant increases in CD4+ cell count (P=.006 and P<.001, respectively), compared with that seen in control patients. These changes were not significant in the 1.5 MIU dose group compared with that in the control patients (P=.105). Side effects that occurred from subcutaneous IL-2 administration were generally low grade, of short duration, and readily managed in an outpatient environment. PMID- 10823762 TI - In vitro activity of human immunodeficiency virus protease inhibitors against Pneumocystis carinii. AB - Since 1996, the introduction of protease inhibitors (PIs) has led to a dramatic decrease of human immunodeficiency virus-related Pneumocystis carinii pneumonia. This effect is clearly due, in large part, to the induction of immune reconstitution by highly active antiretroviral therapy (HAART). However, it is conceivable that PIs had other beneficial effects, including direct activity against Pneumocystis. In this study, the occurrence of specific aspartyl proteases in Pneumocystis is described. These protease targets seemed to be affected in vitro by antiretroviral PIs. These data suggest intriguing implications for the possible antipneumocystis benefit of receiving indinavir, ritonavir, nelfinavir, or saquinavir during HAART. PMID- 10823763 TI - Discontinuation of primary prophylaxis for Pneumocystis carinii pneumonia and toxoplasmic encephalitis in human immunodeficiency virus type I-infected patients: the changes in opportunistic prophylaxis study. AB - A multicenter open, randomized, controlled trial was conducted to determine whether primary prophylaxis for Pneumocystis carinii pneumonia and toxoplasmic encephalitis can be discontinued in patients infected with human immunodeficiency virus type 1 (HIV-1) whose CD4+ T cell counts have increased to >200 cells/mm3 (and who have remained at this level for at least 3 months) as a result of highly active antiretroviral therapy (HAART). Patients were randomized to either the discontinuation arm (i.e., those who discontinued prophylaxis; n=355) or to the continuation arm (n=353); the 2 arms of the study were similar in terms of demographic, clinical, and immunovirologic characteristics. During the median follow-ups of 6.4 months (discontinuation arm) and 6.1 months (continuation arm) and with a total of 419 patient-years, no patient developed P. carinii pneumonia or toxoplasmic encephalitis. The results of this study strongly indicate that primary prophylaxis for P. carinii pneumonia and toxoplasmic encephalitis can be safely discontinued in patients whose CD4+ T cell counts increase to >200 cells/mm3 during HAART. PMID- 10823764 TI - Persistent diarrhea signals a critical period of increased diarrhea burdens and nutritional shortfalls: a prospective cohort study among children in northeastern Brazil. AB - Persistent diarrhea (PD; duration >/=14 days) is a growing part of the global burden of diarrheal diseases. A 45-month prospective cohort study (with illness, nutritional, and microbiologic surveillance) was conducted in a shantytown in northeastern Brazil, to elucidate the epidemiology, nutritional impact, and causes of PD in early childhood (0-3 years of age). A nested case-control design was used to examine children's diarrhea burden and nutritional status before and after a first PD illness. PD illnesses accounted for 8% of episodes and 34% of days of diarrhea. First PD illnesses were preceded by a doubling of acute diarrhea burdens, were followed by further 2.6-3.5-fold increased diarrhea burdens for 18 months, and were associated with acute weight shortfalls. Exclusively breast-fed children had 8-fold lower diarrhea rates than did weaned children. PD-associated etiologic agents included Cryptosporidium, Giardia, enteric adenoviruses, and enterotoxigenic Escherichia coli. PD signals growth shortfalls and increased diarrhea burdens; children with PD merit extended support, and the illness warrants further study to elucidate its prevention, treatment, and impact. PMID- 10823765 TI - A typing system for neisseria gonorrhoeae based on biotinylated oligonucleotide probes to PIB gene variable regions. AB - The porin proteins PIA and PIB of Neisseria gonorrhoeae are serotyping antigens for the serovar classification system and leading candidates for gonococcal vaccine development. Although serotyping has been a useful tool, this method can be insensitive to critical sequence changes in the por gene, including those in surface-exposed variable regions (VRs). A sensitive and specific typing system for N. gonorrhoeae has been developed that uses biotin-labeled oligonucleotide probes with chemiluminescence detection to type PIB gene VRs. The PIB VR types of geographically and temporally diverse gonococcal strains and sexual contact isolates were determined. por VR typing discriminated between most unrelated isolates and provided information about individual VR type that was not apparent from serovar designations. PIB VR typing avoids limited monoclonal antibody availability, interlaboratory variation, and the requirement for culture-based surveillance associated with gonococcal serotyping, and provides useful information about the molecular epidemiology of individual por gene VRs. PMID- 10823766 TI - Vibrio parahaemolyticus infections in the United States, 1973-1998. AB - Vibrio parahaemolyticus infections are associated with consumption of raw or undercooked shellfish, contaminated food, and exposure of wounds to warm seawater. Foodborne outbreaks and sporadic infections from Vibrio species in 4 Gulf Coast states are reported routinely to the Centers for Disease Control and Prevention (CDC). Between 1988 and 1997, 345 sporadic V. parahaemolyticus infections were reported: 59% were gastroenteritis, 34% were wound infections, 5% were septicemia, and 2% were from other exposures. Forty-five percent of patients suffering from these conditions were hospitalized for their infections, and 88% of persons with acute gastroenteritis reported having eaten raw oysters during the week before their illness occurred. Between 1973 and 1998, 40 outbreaks of V. parahaemolyticus infections were reported to the CDC, and these outbreaks included >1000 illnesses. Most of these outbreaks occurred during the warmer months and were attributed to seafood, particularly shellfish. The median attack rate among persons who consumed the implicated seafood was 56%. To prevent V. parahaemolyticus infections, persons should avoid consumption of raw or undercooked shellfish and exposure of wounds to seawater. PMID- 10823767 TI - Two-year study of the protective efficacy of the oral whole cell plus recombinant B subunit cholera vaccine in Peru. AB - The protective efficacy of an oral inactivated whole cell Vibrio cholerae plus recombinant B subunit cholera vaccine was determined against El Tor cholera among Peruvian children and adults (2-65 years old) in a randomized, double-blind manner. Study subjects received 2 doses of vaccine or placebo 2 weeks apart, followed by a booster dose 10 months later. Surveillance for cholera was performed actively, with 2 visits per week to each household, and passively, at a local hospital. Stool samples were collected during diarrhea episodes and were cultured for V. cholerae. A total of 17,799 persons received 2 doses of vaccine or placebo, and 14,997 of these persons received the booster dose. After 2 doses (first surveillance period), V. cholerae biotype O1 was isolated from 17 vaccinees and 16 placebo recipients, demonstrating vaccine efficacy (VE) of -4%. After 3 doses (second surveillance period), V. cholerae O1 was isolated from 13 vaccinees and 32 placebo recipients, demonstrating VE of 61% (95% confidence interval ?CI, 28%-79%). In the second surveillance period, the VE for illness requiring hospitalization was 82% (95% CI, 27%-96%). VE was also higher for persons >15 years old (VE, 72%; 95% CI, 28%-89%). PMID- 10823768 TI - Genetic variability determinants of Helicobacter pylori: influence of clinical background and geographic origin of isolates. AB - Helicobacter pylori has an unusual pattern of genetic variation, which complicates research on this organism. To gain a better understanding of the forces behind this phenomenon, the extent to which recombination and single point mutations affect genetic variability in H. pylori was quantified and the influence of both geographical distance and clinical background were assessed. Site-directed restriction-endonuclease digestion of 2 gene fragments was performed on 168 isolates from Montreal and Berlin. Allelic diversity was found to be much higher for H. pylori than for other bacterial species. This finding is consistent with those of previous studies on H. pylori that were conducted using other techniques. However, nucleotide diversity was within the range reported for other bacterial species. Phylogenetic analysis found no grouping of strains with clinical background or geographical origin. Recombination at a rate that resulted in linkage equilibrium within genes can explain these observations. PMID- 10823769 TI - Immunization with a Pseudomonas aeruginosa elastase peptide reduces severity of experimental lung infections due to P. aeruginosa Or Burkholderia cepacia. AB - Pseudomonas aeruginosa and Burkholderia cepacia produce metalloproteases that effect lung injury. Two epitopes (peptides 15 and 42) previously identified on P. aeruginosa elastase induce the production of antibodies that neutralize protease activity. The effects of immunization with synthetic peptides based on these epitopes on experimental lung infections due to P. aeruginosa or B. cepacia were examined. Rats were immunized with peptides conjugated to keyhole limpet hemocyanin or tetanus toxoid before infection. Immunization with peptide 15 (pep15) resulted in a decrease in total cells and polymorphonuclear leukocytes in bronchoalveolar lavage (BAL) fluid and a 50%-70% decrease in lung histopathologic changes, compared with findings in controls. Immunization with peptide 42 decreased cells in BAL fluid but did not decrease lung pathologic changes. Immunization with pep15 alone was just as effective in protecting against lung injury as immunization with a combination of both peptides. These studies suggest that immunization with pep15 can reduce the severity of lung infections due to P. aeruginosa or B. cepacia. PMID- 10823770 TI - Induction of acute pleural inflammation by Staphylococcus aureus. I. CD4+ T cells play a critical role in experimental empyema. AB - Bacterial empyema is a frequent complication of pneumonia in patients with acquired immunodeficiency syndrome (AIDS). A model of Staphylococcus aureus empyema was developed that closely resembles bacterial empyema in patients infected with human immunodeficiency virus (HIV). Results show a compartmentalized chemokine response in bacterial empyema. The chemokine levels were higher in the pleural compartment than in the peripheral circulation. Polymorphonuclear leukocyte counts, murine GRO-alpha (KC), and macrophage inflammatory protein-2 levels were significantly (P<.001) lower in CD4+ knockout (CD4 KO) mice pleural fluid than in CD4+ wild-type (CD4 WT) mice. The CD4 KO mice had poorer bacterial clearance than CD4 WT mice. During S. aureus infection, interleukin-10 levels increased in the CD4 KO mice, whereas interferon-gamma levels were increased in CD4 WT mice. CD4+ T cell depletion results in a decreased pleural chemokine response, decreased neutrophil influx into pleural space, and impaired bacterial clearance in empyema. PMID- 10823771 TI - Antibody response to the 60-kDa heat-shock protein of Chlamydia pneumoniae in patients with coronary artery disease. AB - Serum specimens from 752 individuals undergoing coronary arteriography were examined for antibodies to Chlamydia pneumoniae. Patients with coronary artery disease (CAD) were more likely to have IgG antibodies to C. pneumoniae than were individuals without CAD (60% vs. 52%; P=.007; odds ratio, 1.8; 95% confidence interval, 1. 17-2.77). Antibodies to recombinant hsp60 of C. pneumoniae were found with nearly the same frequency in patients with CAD and individuals without CAD (29% vs. 30%; P=.751). There was no association between chlamydial hsp60 antibodies and the severity of CAD or a previous myocardial infarction. Patient sera reacted most frequently to C. pneumoniae proteins of 17, 38, 40, 58, and 60/62 kDa. Reactivity to these proteins was not different between patients with and without CAD. Study results indicate that neither antibodies to chlamydial hsp60 nor antibodies to other C. pneumoniae proteins are useful for discriminating between seropositive patients with and without CAD. PMID- 10823772 TI - Molecular evidence for strain dissemination of Penicillium marneffei: an emerging pathogen in Taiwan. AB - From January 1987 through December 1998, Penicillium marneffei infection (23 patients) or colonization (1 patient) was diagnosed in a total of 24 patients in Taiwan. Of these 24 patients, 16 (67%) had AIDS and 20 (83%) had disseminated P. marneffei infection. The majority (63%) of the infections were considered indigenous. The number of cases has increased markedly in recent years, with 17 of the 24 cases diagnosed from 1996 through 1998. Twenty preserved isolates of P. marneffei, recovered from 11 patients treated at National Taiwan University Hospital during the period of January 1996 through December 1998, were studied to determine the epidemiology of P. marneffei infections. Among the 20 isolates, a total of 8 strains (highly related isolates) were identified on the basis of tests for susceptibility to 5 antifungal agents, for chromosomal DNA restriction fragment-length polymorphism types, and for randomly amplified polymorphic DNA patterns. One of the strains (6 isolates) was isolated from 4 patients treated in 1997 and 1998. Strain spreading of P. marneffei may partially contribute to the increased number of infections caused by this organism in immunosuppressed patients in Taiwan. PMID- 10823773 TI - Early detection of aspergillus infection after allogeneic stem cell transplantation by polymerase chain reaction screening. AB - Invasive aspergillosis (IA) has become a major cause of mortality in patients after allogeneic stem cell transplantation. To assess the potential of prospective polymerase chain reaction (PCR) screening for early diagnosis of IA, 84 recipients of an allogeneic stem cell transplant were analyzed with the investigators blinded to clinical and microbiologic data. Of 1193 blood samples analyzed, 169 (14.2%) were positive by PCR. In patients with newly diagnosed IA (n=7), PCR positivity preceded the first clinical signs by a median of 2 days (range, 1-23 days) and preceded clinical diagnosis of IA by a median of 9 days (range, 2-34 days). Pretransplantation IA (relative risk [RR], 2.37), acute graft versus-host disease (RR, 2.75), and corticosteroid treatment (RR, 6.5) were associated with PCR positivity. The PCR assay revealed a sensitivity of 100% (95% confidence interval [CI], 48%-100%) and a specificity of 65% (95% CI, 53%-75%). None of the PCR-negative patients developed IA during the study period. Thus, prospective PCR screening allows for identification of patients at high risk for subsequent onset of IA. PMID- 10823774 TI - Investigation of anti-WI-1 adhesin antibody-mediated protection in experimental pulmonary blastomycosis. AB - Infection with Blastomyces dermatitidis elicits strong antibody responses to the surface adhesin WI-1. The antibodies are directed chiefly against the adhesive domain, a 25-amino-acid repeat. Tandem-repeat-specific monoclonal antibodies (mAbs) were studied for their opsonic activity in vitro and their capacity to adoptively transfer protection in murine experimental blastomycosis. mAbs to WI-1 enhanced binding and entry of B. dermatitidis yeasts into J774. 16 cells but did not enhance killing or growth inhibition of the yeast. Passive transfer of 8 mAbs to WI-1 into 3 different inbred strains of mice also did not improve the course of experimental infection and sometimes worsened it. mu-deficient mice were more resistant to experimental blastomycosis than were intact littermates, and passive transfer of the mAbs into these mice did not protect them against experimental infection. Thus, antibody to WI-1 does not appear to improve the outcome of murine blastomycosis and may enhance the infection. PMID- 10823775 TI - The major surface glycoprotein expression sites of two special forms of rat Pneumocystis carinii differ in structure. AB - Pneumocystis carinii forma specialis carinii has a unique locus called the UCS, which controls surface antigen variation by acting as the expression site for a family of genes encoding isoforms of the major surface glycoprotein (MSG). Every MSG mRNA begins with 380 nucleotides copied from the UCS locus. This UCS leader sequence is necessary for initiation of translation and for protein processing. Given the UCS's importance in P. carinii f. sp. carinii, it was of interest to examine the UCS locus in the related P. carinii f. sp. ratti. The first 380 nucleotides of P. carinii f. sp. ratti MSG mRNAs were 59% identical to the P. carinii f. sp. carinii UCS. However, the DNA encoding the P. carinii f. sp. ratti UCS was not unique in the genome. Instead, only the first 97 nucleotides came from a unique locus and the remainder from >/=7 loci. These data suggest that the two organisms may use different mechanisms to generate surface variation. PMID- 10823776 TI - The impact of placental malaria on gestational age and birth weight. AB - Maternal malaria is associated with reduced birth weight, which is thought to be effected through placental insufficiency, which leads to intrauterine growth retardation (IUGR). The impact of malaria on preterm delivery is unclear. The effects of placental malaria-related changes on birth weight and gestational age were studied in 1177 mothers (and their newborns) from Tanzania. Evidence of malaria infection was found in 75.5% of placental samples. Only massive mononuclear intervillous inflammatory infiltration (MMI) was associated with increased risk of low birth weight (odds ratio ?OR, 4.0). Maternal parasitized red blood cells and perivillous fibrin deposition both were associated independently with increased risk of premature delivery (OR, 3.2; OR, 2.1, respectively). MMI is an important mechanism in the pathogenesis of IUGR in malaria-infected placentas. This study also shows that placental malaria causes prematurity even in high-transmission areas. The impact of maternal malaria on infant mortality may be greater than was thought previously. PMID- 10823777 TI - Anti-merozoite surface protein-1 19-kDa IgG in mother-infant pairs naturally exposed to Plasmodium falciparum: subclass analysis with age, exposure to asexual parasitemia, and protection against malaria. V. The Asembo Bay Cohort Project. AB - The anti-merozoite surface protein-1 19-kDa IgG (anti-MSP119KD) IgG responses of 33 parasitemic infants, aged 6-14 months, were compared with those of their mothers at the time of the infant's delivery and at the time the infants were sampled; the antimalaria protection associated with these responses was also compared. IgG1 and IgG3 were the predominant subclasses. Infants <300 days old and pregnant mothers had the lowest cytophilic-to-noncytophilic IgG ratio. By 300 days of age, the infants had IgG subclass compositions and levels similar to those of their mothers at the same date. Among infants, older infants with only 1 or 2 detected asexual parasitemias had the highest cytophilic-to-noncytophilic IgG ratio and IgG1 levels. IgG1 level was negatively correlated with protection. The findings suggest that the MSP119KD antibody response develops with age, not with multiple experiences with parasitemia, and, thus, that an antimalaria vaccine strategy for pregnant mothers could delay infants' first parasitemias until they are more capable of mounting a favorable anti-MSP119KD response. PMID- 10823778 TI - Proposal for a new inclusive designation for extraintestinal pathogenic isolates of Escherichia coli: ExPEC. PMID- 10823779 TI - Case-control study of risk factors for human infection with a new zoonotic paramyxovirus, Nipah virus, during a 1998-1999 outbreak of severe encephalitis in Malaysia. AB - An outbreak of encephalitis affecting 265 patients (105 fatally) occurred during 1998-1999 in Malaysia and was linked to a new paramyxovirus, Nipah, that infected pigs, humans, dogs, and cats. Most patients were pig farmers. Clinically undetected Nipah infection was noted in 10 (6%) of 166 community-farm controls (persons from farms without reported encephalitis patients) and 20 (11%) of 178 case-farm controls (persons from farms with encephalitis patients). Case patients (persons with Nipah infection) were more likely than community-farm controls to report increased numbers of sick/dying pigs on the farm (59% vs. 24%, P=.001) and were more likely than case-farm controls to perform activities requiring direct contact with pigs (86% vs. 50%, P=.005). Only 8% of case patients reported no contact with pigs. The outbreak stopped after pigs in the affected areas were slaughtered and buried. Direct, close contact with pigs was the primary source of human Nipah infection, but other sources, such as infected dogs and cats, cannot be excluded. PMID- 10823780 TI - Risk factors for Nipah virus infection among abattoir workers in Singapore. AB - During 10-19 March 1999, 11 workers in 1 of 2 Singaporean abattoirs developed Nipah-virus associated encephalitis or pneumonia, resulting in 1 fatality. A case control study was conducted to determine occupational risk factors for infection. Case patients were abattoir A workers who had anti-Nipah IgM antibodies; control subjects were randomly selected abattoir A workers who tested negative for anti Nipah IgM. All 13 case patients versus 26 (63%) of 41 control subjects reported contact with live pigs (P=.01). Swine importation from Malaysian states concurrently experiencing a Nipah virus outbreak was banned on 3 March 1999; on 19 March 1999, importation of Malaysian pigs was banned, and abattoirs were closed. No unusual illnesses among pigs processed during February-March were reported. Contact with live pigs appeared to be the most important risk factor for human Nipah virus infection. Direct contact with live, potentially infected pigs should be minimized to prevent transmission of this potentially fatal zoonosis to humans. PMID- 10823781 TI - Serum neutralizing antibody titers of seropositive chimpanzees immunized with vaccines coformulated with natural fusion and attachment proteins of respiratory syncytial virus. AB - Subunit vaccines formulated with purified fusion proteins from the A2 (PFP-2) or attenuated 248/404 (PFP-3) strains of respiratory syncytial virus (RSV) were evaluated, either alone or in combination with native attachment (G) protein, for their ability to augment serum neutralizing antibody titers in seropositive chimpanzees. The results suggested that combination vaccines enhanced serum neutralizing antibody titers against both laboratory strains and clinical isolates of RSV. When compared with PFP-2 alone, the resultant neutralizing antibody titers after vaccination with PFP-2+Ga protein were significantly elevated against 71% of A strains tested. In a confirmatory experiment, immunization with PFP-3+Ga+Gb proteins resulted in elevated serum neutralizing antibody titers against 86% of A and 50% of B strains tested versus injection with PFP-3 alone. The results suggest that subunit vaccines composed of both PFP and G proteins have more potential than PFP alone to augment neutralizing antibody titers in seropositive recipients. PMID- 10823782 TI - Monocyte interleukin-12 production is inversely related to duration of respiratory failure in respiratory syncytial virus bronchiolitis. AB - The correlation of clinical and immunological parameters with the duration of respiratory failure was investigated to identify factors determining the clinical outcome of respiratory syncytial virus (RSV) bronchiolitis necessitating mechanical ventilation. At initiation of mechanical ventilation in 30 patients with RSV, production of interleukin (IL)-12 and IL-10 was measured in 48-h peripheral blood cell cultures that were stimulated with lipopolysaccharide and interferon-gamma. The ventilation index (VI)-an indicator of respiratory dysfunction that includes partial pressure of arterial CO2, peak airway pressure, and respiratory rate-correlated with the duration of mechanical ventilation (r=.47; P=.013). Age was not associated with the duration of mechanical ventilation. A highly significant inverse correlation was found between the duration of mechanical ventilation and the production of IL-12 at admission (r= .62; P<.001). This correlation was independent of VI. No correlation was found between IL-10 production and the duration of mechanical ventilation. It is hypothesized that low monocyte IL-12 response during initial RSV infection adversely affects clinical outcome of patients with severe RSV bronchiolitis. PMID- 10823783 TI - Outbreak of poliomyelitis in Angola. AB - Between January and June 1999, 1,100 suspected cases of poliomyelitis were reported in Angola. Poliovirus types 3 and 1 were isolated. Patients' ages ranged from 2 months to 14 years. Of the 588 patients whose vaccine status was known, 58 (9.9%) received >4 doses, 216 (36.7%) received 3 or 4 doses, 178 (30.3%) received 1 or 2 doses, and 136 (23.1%) had no history of vaccination. Civil conflict, economic decline, and crowded areas with scarce sanitation and poor water supply are the most important factors implicated in declining rates of routine vaccination, low population immunity, and intense wild poliovirus transmission. The socioeconomic situation and poor roads have created major difficulties for vaccination and surveillance. The Angolan outbreak has serious implications for the global eradication of poliomyelitis. Surveillance of acute flaccid paralysis remains essential in the assessment of strategies for eradication and interventions to interrupt wild poliovirus transmission. PMID- 10823784 TI - Rhinovirus infection induces major histocompatibility complex class I and costimulatory molecule upregulation on respiratory epithelial cells. AB - Human respiratory epithelial cells may act as antigen-presenting cells during respiratory viral infections. In addition to major histocompatibility complex (MHC) molecules, antigen presentation requires participation of costimulatory molecules. Here the authors investigated class I and class II antigens and B7-1 and B7-2 costimulatory molecule expression in human A549 pulmonary epithelial cells and primary bronchial epithelial cells (HBECs) at baseline and after rhinovirus infection. Constitutive expression of MHC class I and B7-1 molecules was observed on both cell types. MHC class I molecules were up-regulated by rhinovirus infection, while B7-1 was up-regulated only on A549 cells. B7-2 molecules were constitutively expressed at a low level and were up-regulated by rhinovirus only on HBECs. Rhinovirus induction of antigen-presenting molecule expression on A549 cells was accompanied by cellular activation in terms of induction of release of the chemokines RANTES and Groalpha. These data show that respiratory epithelium expresses full antigen-presentation machinery and that rhinovirus infection up-regulates this expression. PMID- 10823785 TI - Detection of Kaposi's sarcoma-associated herpesvirus in oral and genital secretions of Zimbabwean women. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) in oral and genital secretions of women may be involved in horizontal and vertical transmission in endemic regions. Nested polymerase chain reaction assays were used to detect KSHV DNA sequences in one-third of oral, vaginal, and cervical specimens and in 42% of peripheral blood mononuclear cell (PBMC) specimens collected from 41 women infected with human immunodeficiency virus type 1 who had Kaposi's sarcoma (KS). KSHV DNA was not detected in specimens from 100 women without KS, 9 of whom were seropositive for KSHV. A positive association was observed between KSHV DNA detection in oral and genital mucosa, neither of which was associated with KSHV DNA detection in PBMC. These data suggest that KSHV replicates in preferred anatomic sites at levels independent of PBMC viremia. Detection of genital-tract KSHV only among relatively immunosuppressed women may provide an explanation for infrequent perinatal transmission of KSHV. PMID- 10823786 TI - Detection of diverse variants of human immunodeficiency virus-1 groups M, N, and O and simian immunodeficiency viruses from chimpanzees by using generic pol and env primer pairs. AB - Human immunodeficiency virus type 1 (HIV-1) infection of humans is the result of independent cross-species transmissions of simian immunodeficiency viruses (SIVcpz) from naturally infected chimpanzees (Pan troglodytes troglodytes) to man. To develop a polymerase chain reaction-based assay capable of detecting members of all major phylogenetic SIVcpz and HIV-1 lineages (groups M, N, and O), primer pairs in conserved pol and env regions were designed. Both primer sets amplified 99%) in amplifying viral sequences from plasma taken from patients infected with HIV-1 group M (n=226) and O (n=17) viruses. PMID- 10823787 TI - Prevalence and prognostic significance of infection with TT virus in patients infected with human immunodeficiency virus. AB - No clear association between human disease and TT virus (TTV) has been documented. A possible pathogenic role of TTV was investigated in patients infected with human immunodeficiency virus (HIV). TTV serum concentrations were estimated in 185 HIV-infected patients by dilution polymerase chain reaction. Of these, 149 (76%) were TTV-positive, compared with 18 (7%) of 252 Danish blood donors (P<. 001). Of the HIV-infected patients who were TTV-positive, 72 (51%) had high TTV viremia (>/=5 times the highest concentration observed among blood donors, i.e., >/=3.5x105 TTV/mL of serum). High TTV viremia was associated with decreased survival (P<.001; relative hazard [RH], 2.0). There was a correlation between lower CD4+ T cell counts and higher TTV titers (P<.01). In a Cox regression model, CD4+ T cell count (P<.001), age (P<.001), HIV viral load (P<.001), beta2 microglobulin (P<.02), and high TTV viremia (P<.01; RH, 1.9) were independent predictors of survival. TTV is suspected to be an opportunistic pathogen with an independent influence on HIV progression. PMID- 10823788 TI - Cytomegalovirus seropositivity and human immunodeficiency virus type 1 RNA levels in individuals with hemophilia. AB - The effect of cytomegalovirus (CMV) seropositivity on the course of human immunodeficiency virus (HIV) type 1 RNA levels and HIV disease progression was assessed in a cohort of 109 hemophilic men infected with HIV-1 for a median of 12.7 years. There was no evidence of higher HIV RNA levels in the first year after HIV seroconversion (P=. 88) or faster rates of increase over infection (P=.20) in the 59 CMV-seropositive individuals than in the CMV-seronegative individuals. In univariate analyses, CMV seropositivity was associated with significantly faster progression to AIDS and death (relative hazards of 1.58 and 2.22, respectively). These effects were unchanged after adjusting for the RNA level, but they were reduced after adjusting for the CD4 cell count, age at seroconversion, and calendar year of follow-up. Thus, the effect of CMV seropositivity on clinical progression remains significant in this cohort but does not appear to be mediated through an increase in HIV RNA levels. PMID- 10823789 TI - Reduction in human immunodeficiency virus type 1 mutations associated with drug resistance after initiating new therapeutic regimens in pretreated patients. AB - The objective of this study was to investigate the effect of initiating a salvage therapy regimen on resistant viruses in heavily treated patients infected with human immunodeficiency virus type 1 (HIV-1). Nineteen patients receiving multiple antiretroviral drugs were tested for HIV-1 mutations associated with drug resistance by using an in-house method at baseline and at weeks 2, 4, and 8 of the salvage-therapy regimen. For the majority of mutations analyzed, the mean number of detectable mutations at baseline was significantly higher than the mean at weeks 2, 4, and 8 of salvage therapy. Introducing new and more potent therapy reduces the number of detectable drug resistance-associated mutations within 8 weeks, and no evidence was found that the new therapy promoted the emergence of novel mutations. PMID- 10823790 TI - High levels of drug-resistant human immunodeficiency virus variants in patients exhibiting increasing CD4+ T cell counts despite virologic failure of protease inhibitor-containing antiretroviral combination therapy. AB - The genotypic mutations associated with indinavir resistance were analyzed in 27 patients who exhibited sustained CD4+ T cell responses to highly active antiretroviral therapy (HAART), despite virologic failure of treatment. After 12 months of HAART, 1 or 2 primary resistance mutations had occurred in 18 (66%) of the patients, and secondary mutations had accumulated in 22 (88%) of the patients. The number and patterns of mutations in the patients who exhibited discrepant responses to HAART did not differ from those observed in patients who exhibited immunologic and virologic failure to therapy. Results indicate that many patients have prolonged immunologic benefits, despite the development of virologic failure and protease inhibitor mutations. The clinical course of this group of patients calls into question the relevance of genotypic resistance and plasma human immunodeficiency virus RNA level as surrogate markers in patients receiving HAART. PMID- 10823791 TI - Thalidomide suppresses Up-regulation of human immunodeficiency virus coreceptors CXCR4 and CCR5 on CD4+ T cells in humans. AB - Concurrent infection in patients with human immunodeficiency virus (HIV) infection increases the expression of HIV coreceptors CXCR4 and CCR5. Thalidomide has beneficial effects in a number of HIV-associated diseases. The effect of thalidomide on CXCR4 and CCR5 expression on CD4+ T cells was determined. Thalidomide produced a dose-dependent inhibition of lipopolysaccharide (LPS) induced up-regulation of CXCR4 and CCR5 in vitro. Antibody to tumor necrosis factor-alpha (TNF-alpha) also attenuated the LPS-induced HIV coreceptor up regulation, which was not further reduced by thalidomide. Thalidomide (400 mg) was orally administered to 6 men, and their blood was stimulated ex vivo with LPS, staphylococcal or mycobacterial antigens, or antibody to CD3 or CD28 cells. All stimuli induced up-regulation of HIV coreceptors, which was reduced after ingestion of thalidomide. Thalidomide may be beneficial in the treatment of intercurrent infections during HIV infection by reducing the up-regulation of CXCR4 and CCR5 expression on CD4+ T cells induced by bacterial and mycobacterial antigens, by a mechanism that involves inhibition of TNF-alpha. PMID- 10823792 TI - Group-specific antibody levels surrounding invasive pneumococcal illness in children infected with human immunodeficiency virus. AB - Pneumococcal antibody levels surrounding systemic pneumococcal illness (SPI) were measured in children infected with human immunodeficiency virus (HIV). Archived serum samples were collected from 28 HIV-infected children who had 34 cases of SPI, caused by pneumococcal groups 4, 6, 9, 14, 19, and 23. Serum samples collected within 23 weeks before and 13 weeks after the SPI were assayed by ELISA for antipneumococcal polysaccharide (PnPs) IgG antibody to 6 representative pneumococcal serotypes. There was a wide range (0. 16-30.80 microg/mL) of pre-SPI anti-PnPs antibody levels to the presumed infecting serotypes, with a geometric mean level of 0.83 microg/mL (n=34). Seventy-six percent of the antibody values were <2.0 microg/mL, and 95% were <5.0 microg/mL. Homologous seroresponses (>/=4 fold rise in anti-PnPs antibody) were detected in only 4 (27%) of 15 paired serum samples. Heterologous, noninfecting group seroresponses were detected frequently (72%) in the paired serum samples from these 4 homologous group seroresponders. PMID- 10823793 TI - Predominance of class II papG allele of Escherichia coli in pyelonephritis in infants with normal urinary tract anatomy. AB - P-fimbrial genotypes of Escherichia coli strains and their possible association with urinary tract abnormalities were studied in infants with pyelonephritis. A total of 153 urinary E. coli strains were analyzed by polymerase chain reaction for class I, II, and III alleles of the pyelonephritis-associated adhesin gene papG. Strains with any class II papG alleles were found significantly more often in infants with normal anatomy and function or in infants with clinically insignificant abnormalities than they were in infants with significant abnormalities (90 of 119 vs. 14 of 34 infants; P<. 001). On the other hand, strains without any papG alleles were found significantly more often in infants with major urinary tract abnormalities (11 of 34 vs. 17 of 119 infants; P=.016). Our genotypic findings indicate that, especially in infants with a normal urinary tract, infection is caused by more-virulent E. coli than is present in infants without a normal urinary tract. This virulence could be due to expression of pyelonephritogenic P fimbriae by an infecting E. coli strain. PMID- 10823794 TI - Escherichia coli O157:H7 strains that express Shiga toxin (Stx) 2 alone are more neurotropic for gnotobiotic piglets than are isotypes producing only Stx1 or both Stx1 and Stx2. AB - Infection with Escherichia coli O157:H7 can lead to hemolytic uremic syndrome (HUS) in some children. Epidemiologic data suggest that Shiga toxin (Stx) 2 producing strains are more frequently associated with HUS than are Stx1-producing strains. Less clear is whether strains that express Stx2 alone are more frequently associated with HUS than strains that express Stx1 and Stx2. Isogenic mutants 933stx1- and 933stx2- were produced from strain 933 (Stx1 and Stx2 producer), and 86-24stx2- was produced from strain 86-24 (Stx2 producer). Neurologic lesions or symptoms developed in 18 (90%) of 20 gnotobiotic piglets orally infected with strain 86-24, in 15 (85%) of 18 infected with mutant 933stx1 , in 9 (31%) of 29 infected with strain 933, in 0 of 5 infected with mutant 86 24stx2-, and in 0 of 6 infected with mutant 933stx2-. It was concluded that strains expressing Stx2 alone are more neurotropic for piglets when fed orally than are those strains expressing Stx1 and 2, whereas Stx1-producing strains induce only diarrhea. It is also conceivable that strains that produce Stx2 may constitute a significant predictive risk factor for HUS in humans. PMID- 10823795 TI - Effect of parenteral antibiotic administration on the establishment of colonization with vancomycin-resistant Enterococcus faecium in the mouse gastrointestinal tract. AB - A mouse model of intestinal colonization with vancomycin-resistant enterococci (VRE) was used to study the effect of different beta-lactam antibiotics on establishment of VRE colonization. A clinical VanB VRE isolate, Enterococcus faecium C68 (102 or 104 cfu), was inoculated by gastric gavage in conjunction with subcutaneous administration of antibiotics. The MIC of ceftriaxone and ticarcillin against VRE strain C68 is >10,000 microg/mL, and the MIC of piperacillin is 1250 microg/mL. Ceftriaxone and ticarcillin-clavulanate treatment groups developed persistently high levels of stool VRE compared with both the saline and the piperacillin-tazobactam (Pip-Taz) groups (P<.008). The level of stool VRE in the Pip-Taz group did not differ from that for the saline group. Thus, in this mouse model, beta-lactam antibiotics with minimal anti-enterococcal activity promoted establishment of high-level VRE colonization, but Pip-Taz (a beta-lactam antibiotic with more potent activity against VRE) did not. PMID- 10823796 TI - An outbreak of Yersinia enterocolitica O:8 infections associated with pasteurized milk. AB - In October 1995, an outbreak of Yersinia enterocolitica O:8 infections occurred in the Upper Valley of Vermont and New Hampshire. Ten patients were identified, median age 9 years (range, 6 months-44 years). Three patients were hospitalized; 1 underwent an appendectomy. Consumption of bottled pasteurized milk from a local dairy was associated with illness (matched odds ratio undefined; lower 95% confidence interval, 1.9). No deficiencies in pasteurization procedures or equipment were detected. Y. enterocolitica O:8 was isolated from 1 raw-milk sample and from a fecal sample from 1 dairy pig. The route of contamination was not determined; this outbreak likely resulted from postpasteurization contamination of milk. Dairy pigs were the most likely source of contamination. Milk bottles were likely contaminated by rinsing with untreated well water prior to filling or by other environmental routes. Educating dairy owners about Y. enterocolitica and postpasteurization contamination is necessary to prevent further outbreaks. PMID- 10823797 TI - An outbreak of Listeria monocytogenes serotype 3a infections from butter in Finland. AB - In February 1999, an outbreak of listeriosis caused by Listeria monocytogenes serotype 3a occurred in Finland. All isolates were identical. The outbreak strain was first isolated in 1997 in dairy butter. This dairy began delivery to a tertiary care hospital (TCH) in June 1998. From June 1998 to April 1999, 25 case patients were identified (20 with sepsis, 4 with meningitis, and 1 with abscess; 6 patients died). Patients with the outbreak strain were more likely to have been admitted to the TCH than were patients with other strains of L. monocytogenes (60% vs. 8%; odds ratio, 17.3; 95% confidence interval, 2.8-136.8). Case patients admitted to the TCH had been hospitalized longer before cultures tested positive than had matched controls (median, 31 vs. 10 days; P=.008). An investigation found the outbreak strain in packaged butter served at the TCH and at the source dairy. Recall of the product ended the outbreak. PMID- 10823798 TI - Immune response to surface protein A of Streptococcus pneumoniae and to high molecular-weight outer membrane protein A of Moraxella catarrhalis in children with acute otitis media. AB - The immune response was evaluated in 11 children with Streptococcus pneumoniae and in 9 children with Moraxella catarrhalis otitis media. The age of the children had a range of 4-32 months. The mean IgG, IgM, and IgA antibody responses to surface protein A (PspA) of S. pneumoniae in sera from children at the acute and convalescent stages were 4864 versus 5831 ng/mL, P<.05, 1075 versus 3752 ng/mL, P<.05, and 67 versus 93 ng/mL, nonsignificant (NS), respectively. The mean IgG, IgM, and IgA antibody responses to the high-molecular-weight outer membrane protein (UspA) of M. catarrhalis in sera from children at acute and convalescent stages were 710 versus 935 mg/mL, NS; 1895 versus 2646 ng/mL, NS; and 121 versus 204 ng/mL, P<.05, respectively. These data show that PspA and UspA are immunogenic in children after otitis media. PMID- 10823799 TI - CD4+ and CD8+ T cells mediate adoptive immunity to aerosol infection of Mycobacterium bovis bacillus Calmette-Guerin. AB - An adoptive-transfer model using recombinase activation gene-deficient (RAG-1-/-) mice was developed to evaluate CD4+ and CD8+ T cell responses to infection with Mycobacterium bovis bacillus Calmette-Guerin (BCG). After receiving immune, unfractionated T cells or T cell subsets isolated by fluorescence-activated cell sorter, the RAG-1-/- mice were exposed to aerosol BCG, and the bacteria load in the infected organs was examined 4 weeks later. Adoptive immunity was expressed more effectively in the spleens than in the lungs. Although CD4+ or unfractionated T cells protected both lungs and spleens, CD8+ T cells conferred significant protection only in the spleens and not in the lungs. The results confirm that in addition to CD4+, CD8+ T cells also play a role in the prevention of bacterial dissemination. This transfer model may be useful for dissecting T cell responses to mycobacterial infection. PMID- 10823800 TI - Detection of active tuberculosis infection by T cell responses to early-secreted antigenic target 6-kDa protein and culture filtrate protein 10. AB - The purified protein derivative (PPD) skin test has no predictive value for tuberculosis (TB) in Mycobacterium bovis bacillus Calmette-Guerin (BCG) vaccinated individuals because of cross-reactive responses to nonspecific constituents of PPD. T cell responses to early-secreted antigenic target 6-kDa protein (ESAT-6) and the newly identified culture filtrate protein 10 (CFP-10), 2 proteins specifically expressed by M. tuberculosis (MTB) but not by BCG strains, were evaluated. Most TB patients responded to ESAT-6 (92%) or CFP-10 (89%). A minority of BCG-vaccinated individuals responded to both ESAT-6 and CFP-10, their history being consistent with latent infection with MTB in the presence of protective immunity. No responses were found in PPD-negative controls. The sensitivity and specificity of the assay were 84% and 100%, respectively, at a cutoff of 300 pg of interferon-gamma/mL. These data indicate that ESAT-6 and CFP 10 are promising antigens for highly specific immunodiagnosis of TB, even in BCG vaccinated individuals. PMID- 10823801 TI - Sex-dependent neutralizing humoral response to Schistosoma mansoni 28GST antigen in infected human populations. AB - The reduction of Schistosoma fecundity observed after experimental vaccination with the Schistosoma mansoni 28-kDa glutathione S-transferase (Sm28GST) antigen has been related to the inhibition of glutathione S-transferase (GST) enzymatic activity by specific antibody. The humoral immune response to the protective antigen Sm28GST and to the epitopes involved in the enzymatic site (amino acid ?aa sequences 10-43 and 190-211) was evaluated in infected individuals before chemotherapy treatment. The capacity of the serum samples to inhibit GST enzymatic activity was assessed. Specific IgG3 response was predominant in the male population with a low intensity of infection and was associated with maximal GST inhibition. In contrast, the neutralizing activity of serum samples from women with a low intensity of infection was correlated with high specific IgA response specifically directed toward the 190-211 epitope. These results strongly support the hypothesis that GST-neutralizing IgG3 and IgA isotypes are sex dependent. The relationship of this specific acquired immune response with the level of intensity of infection is discussed. PMID- 10823803 TI - Reply PMID- 10823802 TI - Demonstration of borna disease virus nucleic acid in a patient with chronic fatigue syndrome. PMID- 10823804 TI - Predictive value of genotypic and phenotypic resistance in 2 nelfinavir experienced patients. PMID- 10823805 TI - Reply PMID- 10823806 TI - To promote circumcision as a preventive measure against human immunodeficiency virus transmission is irresponsible. PMID- 10823807 TI - No protective effect of circumcision on human immunodeficiency virus incidence. PMID- 10823808 TI - Reply PMID- 10823810 TI - Reply PMID- 10823809 TI - Oxidation of low-density lipoproteins by acellular components of Chlamydia pneumoniae. PMID- 10823811 TI - Assessment of specificity of a recombinant 10-kDa protein antigen in differential diagnosis of neurocysticercosis. PMID- 10823812 TI - Reply PMID- 10823813 TI - Worms and pediatric human immunodeficiency virus infection and tuberculosis. PMID- 10823814 TI - Deregulation of glucose transporter 1 and glycolytic gene expression by c-Myc. AB - Unlike normal mammalian cells, which use oxygen to generate energy, cancer cells rely on glycolysis for energy and are therefore less dependent on oxygen. We previously observed that the c-Myc oncogenic transcription factor regulates lactate dehydrogenase A and induces lactate overproduction. We, therefore, sought to determine whether c-Myc controls other genes regulating glucose metabolism. In Rat1a fibroblasts and murine livers overexpressing c-Myc, the mRNA levels of the glucose transporter GLUT1, phosphoglucose isomerase, phosphofructokinase, glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase, and enolase were elevated. c-Myc directly transactivates genes encoding GLUT1, phosphofructokinase, and enolase and increases glucose uptake in Rat1 fibroblasts. Nuclear run-on studies confirmed that the GLUT1 transcriptional rate is elevated by c-Myc. Our findings suggest that overexpression of the c-Myc oncoprotein deregulates glycolysis through the activation of several components of the glucose metabolic pathway. PMID- 10823815 TI - Bile acid induction of cytokine expression by macrophages correlates with repression of hepatic cholesterol 7alpha-hydroxylase. AB - In the studies reported herein, we show that two complementary experimental models: inbred strains of mice (i.e. C57BL/6 and C3H/HeJ), and a differentiated line of rat hepatoma cells (i.e. L35 cells), require the activation of cytokines by monocyte/macrophages to display bile acid negative feedback repression of cholesterol 7alpha-hydroxylase (CYP7A1). Feeding a bile acid-containing atherogenic diet for 3 weeks to C57BL/6 mice led to a 70% reduction in the expression of hepatic CYP7A1 mRNA, whereas no reduction was observed in C3H/HeJ mice. The strain-specific response to repression of CYP7A1 paralleled the activation of hepatic cytokine expression. Studies using cultured THP-1 monocyte/macrophages showed that the hydrophobic bile acid chenodeoxycholate, a well established potent repressor of CYP7A1, induced the expression of mRNAs encoding interleukin 1 (IL-1) and tumor necrosis factor alpha (TNFalpha). In contrast, the hydrophilic bile acid ursodeoxycholate, which does not repress CYP7A1, did not induce cytokine mRNA expression by THP-1 cells. Chenodeoxycholate activation of cytokines by THP-1 cells was blocked by the peroxisome proliferator activated receptor gamma agonist rosiglitazone. The expression of cytokines (e.g. IL-1 and TNFalpha) by THP-1 cells paralleled with the ability of these cells to produce conditioned medium that when added to rat L35 hepatoma cells, repressed CYP7A1. Moreover, rosiglitazone, which blocks cytokine activation by macrophages, also blocked the repression of CYP7A1 normally exhibited by C57BL/6 mice fed the bile acid-containing atherogenic diet. The combined data indicate that the activation of cytokines may mediate CYP7A1 repression caused by feeding mice an atherogenic diet containing bile acids. PMID- 10823816 TI - Decorin binds near the C terminus of type I collagen. AB - Decorin belongs to a family of small leucine-rich proteoglycans that are directly involved in the control of matrix organization and cell growth. Genetic evidence indicates that decorin is required for the proper assembly of collagenous matrices. Here, we sought to establish the precise binding site of decorin on type I collagen. Using rotary shadowing electron microscopy and photoaffinity labeling, we mapped the binding site of decorin protein core to a narrow region near the C terminus of type I collagen. This region is located within the cyanogen bromide peptide fragment alpha1(I) CB6 and is approximately 25 nm from the C terminus, in a zone that coincides with the c(1) band of the collagen fibril d-period. This location is very close to one of the major intermolecular cross-linking sites of collagen heterotrimers. Thus, decorin protein core possesses a unique binding specificity that could potentially regulate collagen fibril stability. PMID- 10823817 TI - Arrestin binding to the G protein-coupled N-formyl peptide receptor is regulated by the conserved "DRY" sequence. AB - Following activation by ligand, the N-formyl peptide receptor (FPR) undergoes processing events initiated by phosphorylation that lead to receptor desensitization and internalization. Our previous results have shown that FPR internalization can occur in the absence of receptor desensitization, suggesting that FPR desensitization and internalization are controlled by distinct mechanisms. More recently, we have provided evidence that internalization of the FPR occurs via a mechanism that is independent of the actions of arrestin, dynamin, and clathrin. In the present report, we demonstrate that stimulation of the FPR with agonist leads to a significant translocation of arrestin-2 from the cytosol to the membrane. Fluorescence microscopy revealed that the translocated arrestin-2 is highly colocalized with the ligand-bound FPR. A D71A mutant FPR, which does not undergo activation or phosphorylation in response to ligand, did not colocalize with arrestin-2. Surprisingly, an R123G mutant FPR, which does not bind G protein but does become phosphorylated and subsequently internalized, also did not bind arrestin. These results indicate that arrestin binding is not required for FPR internalization and demonstrate for the first time that a common motif, the conserved "DRY" domain of G protein-coupled receptors, is essential for phosphorylation-dependent arrestin binding, as well as G protein activation. PMID- 10823818 TI - Characterization of the recombinant IKK1/IKK2 heterodimer. Mechanisms regulating kinase activity. AB - Nuclear factor kappa B (NF-kappaB) is a ubiquitous, inducible transcription factor that regulates the initiation and progression of immune and inflammatory stress responses. NF-kappaB activation depends on phosphorylation and degradation of its inhibitor protein, IkappaB, initiated by an IkappaB kinase (IKK) complex. This IKK complex includes a catalytic heterodimer composed of IkappaB kinase 1 (IKK1) and IkappaB kinase 2 (IKK2) as well as a regulatory adaptor subunit, NF kappaB essential modulator. To better understand the role of IKKs in NF-kappaB activation, we have cloned, expressed, purified, and characterized the physiological isoform, the rhIKK1/rhIKK2 heterodimer. We compared its kinetic properties with those of the homodimers rhIKK1 and rhIKK2 and a constitutively active rhIKK2 (S177E, S181E) mutant. We demonstrate activation of these recombinantly expressed IKKs by phosphorylation during expression in a baculoviral system. The K(m) values for ATP and IkappaBalpha peptide for the rhIKK1/rhIKK2 heterodimer are 0.63 and 0.60 micrometer, respectively, which are comparable to those of the IKK2 homodimer. However, the purified rhIKK1/rhIKK2 heterodimer exhibits the highest catalytic efficiency (k(cat)/K(m)) of 47.50 h( 1) micrometer(-1) using an IkappaBalpha peptide substrate compared with any of the other IKK isoforms, including rhIKK2 (17.44 h(-1) micrometer(-1)), its mutant rhIKK2 (S177E, S181E, 1.18 h(-1) micrometer(-1)), or rhIKK1 (0.02 h(-1) micrometer(-1)). Kinetic analysis also indicates that, although both products of the kinase reaction, ADP and a phosphorylated IkappaBalpha peptide, exhibited competitive inhibitory kinetics, only ADP with the low K(i) of 0.77 micrometer may play a physiological role in regulation of the enzyme activity. PMID- 10823819 TI - Fast inactivation of voltage-dependent calcium channels. A hinged-lid mechanism? AB - We recently described domains II and III as important determinants of fast, voltage-dependent inactivation of R-type calcium channels (Spaetgens, R. L., and Zamponi, G. W. (1999) J. Biol. Chem. 274, 22428-22438). Here we examine in greater detail the structural determinants of inactivation using a series of chimeras comprising various regions of wild type alpha(1C) and alpha(1E) calcium channels. Substitution of the II S6 and/or III S6 segments of alpha(1E) into the alpha(1C) backbone resulted in rapid inactivation rates that closely approximated those of wild type alpha(1E) channels. However, neither individual or combined substitution of the II S6 and III S6 segments could account for the 60 mV more negative half-inactivation potential seen with wild type alpha(1E) channels, indicating that the S6 regions contribute only partially to the voltage dependence of inactivation. Interestingly, the converse replacement of alpha(1E) S6 segments of domains II, III, or II+III with those of alpha(1C) was insufficient to significantly slow inactivation rates. Only when the I-II linker region and the domain II and III S6 regions of alpha(1E) were concomitantly replaced with alpha(1C) sequence could inactivation be abolished. Conversely, introduction of the alpha(1E) domain I-II linker sequence into alpha(1C) conferred alpha(1E)-like inactivation rates, indicating that the domain I-II linker is a key contributor to calcium channel inactivation. Overall, our data are consistent with a mechanism in which inactivation of voltage-dependent calcium channels may occur via docking of the I-II linker region to a site comprising, at least in part, the domain II and III S6 segments. PMID- 10823820 TI - Tyrosine phosphorylation of paxillin alpha is involved in temporospatial regulation of paxillin-containing focal adhesion formation and F-actin organization in motile cells. AB - Temporal and spatial regulation of actin-based cytoskeletal organization and focal adhesion formation play an essential role in cell migration. Here, we show that tyrosine phosphorylation of a focal adhesion protein, paxillin, crucially participates in these regulations. We found that tyrosine phosphorylation of paxillin was a prominent event upon integrin activation during epithelial mesenchymal trans-differentiation and cell migration. Four major tyrosine phosphorylation sites were identified, and two of them were highly inducible upon integrin activation. Paxillin exhibits three distinct subcellular localizations as follows: localization along the cell periphery colocalized with circumferential actin meshworks, macroaggregation at focal adhesions connected to actin stress fibers, and diffuse cytoplasmic distribution. Tyrosine phosphorylation of paxillin localized at the cell periphery and focal adhesions was shown using phosphorylation site-specific antibodies. Mutations in the phosphorylation sites affected the peripheral localization of paxillin and paxillin-containing focal adhesion formation during cell migration and cell-cell collision, accompanied by altered actin organizations. Our analysis indicates that phosphorylation of multiple tyrosines in paxillin alpha is necessary for the proper function of paxillin and is involved in the temporospatial regulation of focal adhesion formation and actin cytoskeletal organization in motile cells. PMID- 10823821 TI - Inhibition of death receptor-mediated gene induction by a cycloheximide-sensitive factor occurs at the level of or upstream of Fas-associated death domain protein (FADD). AB - In HeLa cells, induction of apoptosis and nuclear factor kappaB (NF-kappaB) activation initiated by TRAIL/Apo2L or the agonistic Apo1/Fas-specific monoclonal antibody anti-APO-1 require the presence of cycloheximide (CHX). Inhibition of caspases prevented TRAIL/anti-APO-1-induced apoptosis, but not NF-kappaB activation, indicating that both pathways bifurcate upstream of the receptor proximal caspase-8. Under these conditions, TRAIL and anti-APO-1 up-regulated the expression of the known NF-kappaB targets interleukin-6, cellular inhibitor of apoptosis 2 (cIAP2), and TRAF1 (TRAF, tumor necrosis factor receptor-associate factor). In the presence of CHX, the stable overexpression of a deletion mutant of the Fas-associated death domain molecule FADD comprising solely the death domain of the molecule but lacking its death effector domain (FADD-(80-208)) led to the same response pattern as TRAIL or anti-APO-1 treatment. Moreover, the ability of death receptors to induce NF-kappaB activation was drastically reduced in a FADD-deficient Jurkat cell line. TRAIL-, anti-APO-1-, and FADD-(80-208) initiated gene induction was blocked by a dominant-negative mutant of TRAF2 or the p38 kinase inhibitor SB203580, similar to tumor necrosis factor receptor-1 induced NF-kappaB activation. CHX treatment rapidly down-regulated endogenous cFLIP protein levels, and overexpression of cellular FLICE inhibitory protein (cFLIP) inhibited death receptor-induced NF-kappaB activation. Thus, a novel functional role of cFLIP as a negative regulator of gene induction by death receptors became apparent. PMID- 10823822 TI - Thioredoxin facilitates the induction of heme oxygenase-1 in response to inflammatory mediators. AB - Heme oxygenase (HO)-1 is a stress response protein that is regulated by oxidative stress. HO-1 catalyzes the generation of biliverdin, carbon monoxide, and iron from heme. Lipopolysaccharide (LPS) and interleukin (IL)-1beta induce HO-1 through the binding of nuclear proteins to AP-1 motifs in enhancer regions upstream from the transcription start site. The DNA binding activity of AP-1 proteins depends on the reduction of cysteines in their DNA-binding domains. We found that agents that disrupt free sulfhydryl groups abolish AP-1 binding activity in nuclear proteins obtained from rat aortic smooth muscle cells and macrophages stimulated with IL-1beta or LPS. Thioredoxin (TRX) may regulate the redox status of nuclear transcription factors in response to oxidative stimuli, thus we determined the role of TRX in the physiologic regulation of HO-1. TRX underwent nuclear translocation in cells stimulated with IL-1beta and LPS. We transfected macrophages with a heterologous promoter construct containing two AP 1 sites from an upstream enhancer region in the HO-1 promoter. Recombinant TRX induced promoter activity to a level analogous to that induced by LPS, and this TRX response was abolished by mutation of the AP-1 sites. An inhibitor of TRX reductase, used to prevent TRX translocation in the reduced state, decreased HO-1 induction by IL-1beta and LPS. These data provide the first evidence that TRX contributes to the induction of HO-1 by inflammatory mediators. PMID- 10823823 TI - Cleavage of the actin-capping protein alpha -adducin at Asp-Asp-Ser-Asp633-Ala by caspase-3 is preceded by its phosphorylation on serine 726 in cisplatin-induced apoptosis of renal epithelial cells. AB - Decreased phosphorylation of focal adhesion kinase and paxillin is associated with loss of focal adhesions and stress fibers and precedes the onset of apoptosis (van de Water, B., Nagelkerke, J. F., and Stevens, J. L. (1999) J. Biol. Chem. 274, 13328-13337). The cortical actin cytoskeletal network is also lost during apoptosis, yet little is known about the temporal relationship between altered phosphorylation of proteins that are critical in the regulation of this network and their potential cleavage by caspases during apoptosis. Adducins are central in the cortical actin network organization. Cisplatin caused apoptosis of renal proximal tubular epithelial cells, which was associated with the cleavage of alpha-adducin into a 74-kDa fragment; this was blocked by a general caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone (z VAD-fmk). Hemagglutinin-tagged human alpha-adducin was cleaved into a similar 74 kDa fragment by caspase-3 in vitro but not by caspase-6 or -7. Asp-Arg-Val Asp(29)-Glu, Asp-Ile-Val-Asp(208)-Arg, and Asp-Asp-Ser-Asp(633)-Ala were identified as the principal caspase-3 cleavage sites; Asp-Asp-Ser-Asp(633)-Ala was key in the formation of the 74-kDa fragment. Cisplatin also caused an increased phosphorylation of alpha-adducin and gamma-adducin in the MARCKS domain that preceded alpha-adducin cleavage and was associated with loss of adducins from adherens junctions; this was not affected by z-VAD-fmk. In conclusion, the data support a model in which increased phosphorylation of alpha-adducin due to cisplatin leads to dissociation from the cytoskeleton, a situation rendered irreversible by caspase-3-mediated cleavage of alpha-adducin at Asp-Asp-Ser Asp(633)-Ala. PMID- 10823824 TI - The role of high molecular weight kininogen and prothrombin as cofactors in the binding of factor XI A3 domain to the platelet surface. AB - We have reported that prothrombin (1 microm) is able to replace high molecular weight kininogen (45 nm) as a cofactor for the specific binding of factor XI to the platelet (Baglia, F. A., and Walsh, P. N. (1998) Biochemistry 37, 2271-2281). We have also determined that prothrombin fragment 2 binds to the Apple 1 domain of factor XI at or near the site where high molecular weight kininogen binds. A region of 31 amino acids derived from high molecular weight kininogen (HK31-mer) can also bind to factor XI (Tait, J. F., and Fujikawa, K. (1987) J. Biol. Chem. 262, 11651-11656). We therefore investigated the role of prothrombin fragment 2 and HK31-mer as cofactors in the binding of factor XI to activated platelets. Our experiments demonstrated that prothrombin fragment 2 (1 microm) or the HK31-mer (8 microm) are able to replace high molecular weight kininogen (45 nm) or prothrombin (1 microm) as cofactors for the binding of factor XI to the platelet. To localize the platelet binding site on factor XI, we used mutant full-length recombinant factor XI molecules in which the platelet binding site in the Apple 3 domain was altered by alanine scanning mutagenesis. The recombinant factor XI with alanine substitutions at positions Ser(248), Arg(250), Lys(255), Leu(257), Phe(260), or Gln(263) were defective in their ability to bind to activated platelets. Thus, the interaction of factor XI with platelets is mediated by the amino acid residues Ser(248), Arg(250), Lys(255), Leu(257), Phe(260), and Gln(263) within the Apple 3 domain. PMID- 10823825 TI - Superoxide-induced stimulation of protein kinase C via thiol modification and modulation of zinc content. AB - We investigated the effects of mild oxidation on protein kinase C (PKC) using the xanthine/xanthine oxidase system of generating superoxide. Exposure of various PKC preparations to superoxide stimulated the autonomous activity of PKC. Similarly, thiol oxidation increased autonomous PKC activity, consistent with the notion that superoxide stimulates PKC via thiol oxidation. The superoxide-induced stimulation of PKC activity was partially reversed by reducing agents, suggesting that disulfide bond formation contributed to the oxidative stimulation of PKC. In addition, superoxide increased the autonomous activity of the alpha, beta(II), epsilon, and zeta PKC isoforms, all of which contain at least one cysteine-rich region. Taken together, our observations suggested that superoxide interacts with PKC at the cysteine-rich region, zinc finger motif of the enzyme. Therefore, we examined the effects of superoxide on this region by testing the hypothesis that superoxide stimulates PKC by promoting the release of zinc from PKC. We found that a zinc chelator stimulated the autonomous activity of PKC and that superoxide induced zinc release from an PKC-enriched enzyme preparation. In addition, oxidized PKC contained significantly less zinc than reduced PKC. Finally, we have isolated a persistent, autonomously active PKC by DEAE-cellulose column chromatography from hippocampal slices incubated with superoxide. Taken together, these data suggest that superoxide stimulates autonomous PKC activity via thiol oxidation and release of zinc from cysteine-rich region of PKC. PMID- 10823826 TI - The lipopolysaccharide-activated toll-like receptor (TLR)-4 induces synthesis of the closely related receptor TLR-2 in adipocytes. AB - The central regulatory role of the adipocyte in whole body energy homeostasis is well established. However, recent findings suggest that preadipocytes and adipocytes may play an important physiological role in the regulation of both the innate and adaptive immune response. To systematically characterize the molecular machinery of the adipocyte that mediates the recognition of pathogens, we have focused our analysis on the recently identified Toll-like receptors (TLRs). These receptors have been implicated as mediators of the cellular response to bacterial lipopolysacharides (LPSs). Here, we report the cloning and functional characterization of mouse TLR-2 from 3T3-L1 adipocytes. TLR-2 synthesis is strongly induced in the adipocyte by LPS, TNFalpha, and the yeast cell wall extract zymosan. TLR-2 undergoes a lengthy intracellular maturation process with a half-life of exit from the ER of approximately 3 h. Furthermore, LPS treatment of adipocytes results in dramatic changes at the level of gene expression, including the synthesis of a distinct set of secretory proteins such as interleukin-6. Our studies demonstrate the presence of a fully intact pathway of innate immunity in the adipocyte that can be activated by LPS binding to the cell surface and results in the secretion of immunomodulatory molecules. PMID- 10823827 TI - Primary structure, genomic organization, and functional and electrogenic characteristics of human system N 1, a Na+- and H+-coupled glutamine transporter. AB - We have cloned the human Na(+)- and H(+)-coupled amino acid transport system N (hSN1) from HepG2 liver cells and investigated its functional characteristics. Human SN1 protein consists of 504 amino acids and shows high homology to rat SN1 and rat brain glutamine transporter (GlnT). When expressed in mammalian cells, the transport function of human SN1 could be demonstrated with glutamine as the substrate in the presence of LiCl (instead of NaCl) and cysteine. The transport activity was saturable, pH-sensitive, and specific for glutamine, histidine, asparagine, and alanine. Analysis of Li(+) activation kinetics showed a Li(+):glutamine stoichiometry of 2:1. When expressed in Xenopus laevis oocytes, the transport of glutamine or asparagine via human SN1 was associated with inward currents under voltage-clamped conditions. The transport function, monitored as glutamine- or asparagine-induced currents, was saturable, Na(+)-dependent, Li(+) tolerant, and pH-sensitive. The transport cycle was associated with the involvement of more than one Na(+) ion. Uptake of asparagine was directly demonstrable in these oocytes by using radiolabeled substrate, and this uptake was inhibited by membrane depolarization. In addition, simultaneous measurement of asparagine influx and charge influx in the same oocyte yielded an asparagine:charge ratio of 1. These data suggest that SN1 mediates the influx of two Na(+) and one amino acid substrate per transport cycle coupled to the efflux of one H(+), rendering the transport process electrogenic. PMID- 10823828 TI - 7The yeast mRNA-binding protein Npl3p interacts with the cap-binding complex. AB - A number of RNA-binding proteins are associated with mRNAs in both the nucleus and the cytoplasm. One of these, Npl3p, is a heterogeneous nuclear ribonucleoprotein-like protein with some similarity to SR proteins and is essential for growth in the yeast S. cerevisiae. Temperature-sensitive alleles have defects in the export of mRNA out of the nucleus (1). In this report, we define a genetic relationship between NPL3 and the nonessential genes encoding the subunits of the cap-binding complex (CBP80 and CBP20). Deletion of CBP80 or CBP20 in combination with certain temperature-sensitive npl3 mutant alleles fail to grow and thus display a synthetic lethal relationship. Further evidence of an interaction between Npl3p and the cap-binding complex was revealed by co immunoprecipitation experiments; Cbp80p and Cbp20p specifically co-precipitate with Npl3p. However, the interaction of Npl3p with Cbp80p depends on both the presence of Cbp20p and RNA. In addition, we show that Cbp80p is capable of shuttling between the nucleus and the cytoplasm in a manner dependent on the ongoing synthesis of RNA. Taken together, these data support a model whereby mRNAs are co-transcriptionally packaged by proteins including Npl3p and cap binding complex for export out of the nucleus. PMID- 10823829 TI - Activation of Stat3 in v-Src-transformed fibroblasts requires cooperation of Jak1 kinase activity. AB - Signal transducers and activators of transcription (STATs) are latent cytoplasmic transcription factors that transduce signals from the cell membrane to the nucleus upon activation by tyrosine phosphorylation. Several protein-tyrosine kinases can induce phosphorylation of STATs in cells, including Janus kinase (JAK) and Src family kinases. One STAT family member, Stat3, is constitutively activated in Src-transformed NIH3T3 cells and is required for cell transformation. However, it is not entirely clear whether Src kinase can phosphorylate Stat3 directly or through another pathway, such as JAK family kinases. To address this question, we investigated the phosphorylation of STATs in baculovirus-infected Sf-9 insect cells in the presence of Src. Our results show that Src can tyrosine-phosphorylate Stat1 and Stat3 but not Stat5 in this system. The phosphorylated Stat1 and Stat3 proteins are functionally activated, as measured by their abilities to specifically bind DNA oligonucleotide probes. In addition, the JAK family member Jak1 efficiently phosphorylates Stat1 but not Stat3 in Sf-9 cells. By contrast, we observe that AG490, a JAK family-selective inhibitor, and dominant negative Jak1 protein can significantly inhibit Stat3 induced DNA binding activity as well as Stat3-mediated gene activation in NIH3T3 cells. Furthermore, wild-type or kinase-inactive platelet-derived growth factor receptor enhances Stat3 activation by v-Src, consistent with the receptor serving a scaffolding function for recruitment and activation of Stat3. Our results demonstrate that Src kinase is capable of activating STATs in Sf-9 insect cells without expression of JAK family members; however, Jak1 and platelet-derived growth factor receptor are required for maximal Stat3 activation by Src kinase in mammalian cells. Based on these findings, we propose a model in which Jak1 serves to recruit Stat3 to a receptor complex with Src kinase, which in turn directly phosphorylates and activates Stat3 in Src-transformed fibroblasts. PMID- 10823830 TI - Involvement of STAT-1 and ets family members in interferon-gamma induction of CD40 transcription in microglia/macrophages. AB - Cluster of differentiation (CD)-40 is a cell surface receptor belonging to the tumor necrosis factor receptor family that plays a critical role in the regulation of immune responses. We have previously shown that the cytokine interferon (IFN)-gamma induces CD40 expression in microglia. Herein, we have elucidated the molecular mechanisms underlying IFN-gamma induction of CD40 gene expression in microglia/macrophages. IFN-gamma up-regulates CD40 expression at the transcriptional level, and this regulation involves the STAT-1alpha transcription factor. Microglia from STAT-1alpha-deficient mice were refractive to IFN-gamma induction of CD40 expression, illustrating the importance of STAT 1alpha in this response. Functional analysis of the CD40 promoter indicates that two gamma activated sequence elements as well as two Ets elements are involved in IFN-gamma induction of CD40 promoter activity. STAT-1alpha binds to the gamma activated sequence elements, whereas PU.1 and/or Spi-B bind to the Ets elements. The expression of PU.1 and Spi-B, in conjuction with STAT-1alpha activation, correlates with IFN-gamma inducibility of CD40 expression. Collectively, our data demonstrate the involvement of STAT-1alpha, PU.1, and Spi-B in IFN-gamma induction of CD40 gene expression in cells of the macrophage lineage. PMID- 10823831 TI - Hypoxia inducible factor-alpha binding and ubiquitylation by the von Hippel Lindau tumor suppressor protein. AB - The von Hippel-Lindau tumor suppressor protein (pVHL) has emerged as a key factor in cellular responses to oxygen availability, being required for the oxygen dependent proteolysis of alpha subunits of hypoxia inducible factor-1 (HIF). Mutations in VHL cause a hereditary cancer syndrome associated with dysregulated angiogenesis, and up-regulation of hypoxia inducible genes. Here we investigate the mechanisms underlying these processes and show that extracts from VHL deficient renal carcinoma cells have a defect in HIF-alpha ubiquitylation activity which is complemented by exogenous pVHL. This defect was specific for HIF-alpha among a range of substrates tested. Furthermore, HIF-alpha subunits were the only pVHL-associated proteasomal substrates identified by comparison of metabolically labeled anti-pVHL immunoprecipitates from proteosomally inhibited cells and normal cells. Analysis of pVHL/HIF-alpha interactions defined short sequences of conserved residues within the internal transactivation domains of HIF-alpha molecules sufficient for recognition by pVHL. In contrast, while full length pVHL and the p19 variant interact with HIF-alpha, the association was abrogated by further N-terminal and C-terminal truncations. The interaction was also disrupted by tumor-associated mutations in the beta-domain of pVHL and loss of interaction was associated with defective HIF-alpha ubiquitylation and regulation, defining a mechanism by which these mutations generate a constitutively hypoxic pattern of gene expression promoting angiogenesis. The findings indicate that pVHL regulates HIF-alpha proteolysis by acting as the recognition component of a ubiquitin ligase complex, and support a model in which its beta domain interacts with short recognition sequences in HIF-alpha subunits. PMID- 10823832 TI - Thermodynamic stability of HLA-B*2705. Peptide complexes. Effect of peptide and major histocompatibility complex protein mutations. AB - Designing synthetic vaccines from class I major histocompatibility complex (MHC) binding antigenic peptides requires not only knowledge of the binding affinity of the designed peptide but also predicting the stability of the formed MHC-peptide complex. In order to better investigate structure-stability relationships, we have determined by circular dichroism spectroscopy the thermal stability of a class I MHC protein, HLA-B*2705, in complex with a set of 39 singly substituted peptide analogues. The influence of two anchoring side chains (P3 and P9) was studied by peptide mutation and appropriate site-directed mutagenesis of the HLA B*2705 binding groove. The side chain at P9 is clearly the one that contributes the most to the thermal stability of the MHC-peptide complexes, as destabilization up to 25 degrees C are obtained after P9 mutation. Interestingly, structure-stability relationships do not fully mirror structure-binding relationships. As important as the C-terminal side chain are the terminal ammonium and carboxylate groups. Removal of a single H-bond between HLA-B27 and the terminal peptide moieties results in thermal destabilization up to 10 degrees C. Depending on the bound peptide and the location of the deleted H-bond, the decrease in the thermal stability of the corresponding complex is quantitatively different. The present study suggests that any peptidic amino acid at positions 3 and 9 promotes refolding of the B27-peptide complex. Once the complex is formed, the C-terminal side chain seems to play an important role for maintaining a stable complex. PMID- 10823833 TI - Transgenic reexpression of GLUT1 or GLUT2 in pancreatic beta cells rescues GLUT2 null mice from early death and restores normal glucose-stimulated insulin secretion. AB - GLUT2-null mice are hyperglycemic, hypoinsulinemic, hyperglucagonemic, and glycosuric and die within the first 3 weeks of life. Their endocrine pancreas shows a loss of first phase glucose-stimulated insulin secretion (GSIS) and inverse alpha to beta cell ratio. Here we show that reexpression by transgenesis of either GLUT1 or GLUT2 in the pancreatic beta cells of these mice allowed mouse survival and breeding. The rescued mice had normal-fed glycemia but fasted hypoglycemia, glycosuria, and an elevated glucagon to insulin ratio. Glucose tolerance was, however, normal. In vivo, insulin secretion assessed following hyperglycemic clamps was normal. In vitro, islet perifusion studies revealed that first phase of insulin secretion was restored as well by GLUT1 or GLUT2, and this was accompanied by normalization of the glucose utilization rate. The ratio of pancreatic insulin to glucagon and volume densities of alpha to beta cells were, however, not corrected. These data demonstrate that 1) reexpression of GLUT1 or GLUT2 in beta cells is sufficient to rescue GLUT2-null mice from lethality, 2) GLUT1 as well as GLUT2 can restore normal GSIS, 3) restoration of GSIS does not correct the abnormal composition of the endocrine pancreas. Thus, normal GSIS does not depend on transporter affinity but on the rate of uptake at stimulatory glucose concentrations. PMID- 10823834 TI - The recruitment of the interleukin-1 (IL-1) receptor-associated kinase (IRAK) into focal adhesion complexes is required for IL-1beta -induced ERK activation. AB - The interleukin-1 (IL-1) receptor colocalizes with focal adhesion complexes (FACs), actin-enriched structures involved in cell adhesion and signaling in fibroblasts and chondrocytes. The colocalization of FACs and IL-1 receptors has been implicated in the restriction of IL-1 signaling transduction to ERK; however, the mechanism of this restriction and the requirement of IL-1 receptor associated proteins have not been characterized. We determined if the association kinetics of the interleukin-1 receptor-associated kinase (IRAK) colocalizes with FACs and the requirement for IRAK in IL-1-dependent ERK activation. Human gingival fibroblasts were incubated with collagen-coated beads to induce the assembly of FACs at sites of cell-bead contact. Immunoblot analysis of bead isolated FACs showed a time-dependent assembly of the focal adhesion proteins beta-actin, vinculin, and talin, which was blocked by the actin monomer sequestering toxin latrunculin B. Although no IRAK was isolated with FACs from unstimulated cells, phosphorylated IRAK was transiently associated with FACs isolated from IL-1beta-stimulated fibroblasts. Fibroblasts plated on tissue culture plastic (which permitted the formation of focal adhesions) showed phosphorylation of ERK, JNK, and p38. Cells plated on poly-l-lysine (to prevent the formation of focal adhesions) showed activation only of JNK and p38. ERK activation was partially restored by incubating cells plated on poly-l-lysine with collagen-coated beads before IL-1 stimulation. Cells treated with latrunculin B or swinholide A, which caused a progressive depolymerization of actin filaments, showed a reduction or elimination of IL-1-induced ERK activation, respectively. Fibroblasts electroinjected with a mouse monoclonal anti-IRAK antibody to block the recruitment of IRAK into FACs failed to activate ERK after IL-1 treatment, indicating that FAC-associated IRAK is required for the activation of ERK. These data indicate that the integrity of actin filament arrays and the recruitment of IRAK into focal adhesions are involved in the restriction of IL-1 signaling to ERK. PMID- 10823835 TI - Functional coupling of human beta 3-adrenoreceptors to the KvLQT1/MinK potassium channel. AB - The slow component of the delayed rectifier potassium current (IKs) plays an important role during repolarization in the human heart. Life-threatening arrhythmias can be triggered by sympathetic stimulation, presumably acting on IKs. The ion channel responsible for the IKs current is made of two proteins, the KvLQT1 protein and the MinK protein. In this study, we investigated the effects of adrenergic stimulation on the KvLQT1/MinK channel by coexpressing KvLQT1/MinK channels with the human beta(3)-adrenoreceptor subunit heterologously in Xenopus oocytes. Western blot experiments revealed that beta(3)-adrenoreceptor proteins appear in the cell membrane of Xenopus oocytes, when the corresponding cRNA was injected. In electrophysiological measurements we found that stimulation with the beta-adrenergic agonist isoproterenol increased the current amplitude of the beta(3)/KvLQT1/MinK complex up to 237% with an ED(50) of 8 nm, a value similar to that found on IKs in guinea pig cardiomyocytes. When oocytes with beta(3)/KvLQT1/MinK were preincubated with cholera toxin (2 microg/ml), an activator of G(S) proteins, the basal current amplitude of the beta(3)/KvLQT1/MinK complex was increased 3.1-fold, and the current amplitude increase by isoproterenol was drastically reduced, indicating that the signal transduction cascade was mediated via G(s) proteins. The knowledge about functional coupling of the human beta(3)-adrenoreceptor to KvLQT1/MinK channels reveals interesting aspects about the genesis and therapy of arrhythmias. PMID- 10823836 TI - Regulation of transporter associated with antigen processing by phosphorylation. AB - The ATP-binding cassette transporter associated with antigen processing (TAP) is required for transport of antigenic peptides, generated by proteasome complexes in the cytoplasm, into the lumen of the endoplasmic reticulum where assembly with major histocompatibility complex class I molecules takes place. The TAP transporter is a heterodimer of TAP1 and TAP2. Here we show that both TAP1 and TAP2 are phosphorylated under physiological conditions. Phosphorylation induces formation of high molecular weight TAP complexes that contain TAP1, TAP2, tapasin, and class I heterodimers. In addition, a 43-kDa phosphoprotein, which appears to be a kinase, is contained in the phosphorylated TAP-containing complexes. Phosphorylated TAP complexes are able to bind peptides and ATP, however, they are not capable of transporting peptides. After de-phosphorylation, TAP complexes regain the ability to transport peptides. Interestingly, phosphorylation levels of TAP complexes induced by viral infection inversely correlates with a significant reduction in TAP-dependent peptide transport activity. Enhanced TAP phosphorylation appears to be one of several strategies that viruses have exploited to better escape from host immune surveillance. These results demonstrate that major histocompatibility complex class I antigen processing and presentation is modulated by reversible TAP phosphorylation, and implicate the importance of TAP phosphorylation in the regulation of cytotoxic immune response. PMID- 10823837 TI - YLL031c belongs to a novel family of membrane proteins involved in the transfer of ethanolaminephosphate onto the core structure of glycosylphosphatidylinositol anchors in yeast. AB - MCD4 and GPI7 are important for the addition of glycosylphosphatidylinositol (GPI) anchors to proteins in the yeast Saccharomyces cerevisiae. Mutations in these genes lead to a reduction of GPI anchoring and cell wall fragility. Gpi7 mutants accumulate a GPI lipid intermediate of the structure Manalpha1-2[NH(2) (CH(2))(2)-PO(4)-->]Manalpha1-2Manalpha 1-6[NH(2)-(C H(2))(2)-PO(4)-->]Manalpha1 4GlcNalpha1-6[acyl-->]inositol-P O(4)-lipi d, which, in comparison with the complete GPI precursor lipid CP2, lacks an HF-sensitive side chain on the alpha1 6-linked mannose. In contrast, mcd4-174 accumulates only minor amounts of abnormal GPI intermediates. Here we investigate whether YLL031c, an open reading frame predicting a further homologue of GPI7 and MCD4, plays any role in GPI anchoring. YLL031c is an essential gene. Its depletion results in a reduction of GPI anchor addition to GPI proteins as well as to cell wall fragility. YLL031c depleted cells accumulate GPI intermediates with the structures Manalpha1 2Manalpha1-2Manalpha1-6[NH(2)-(CH(2))(2)-PO( 4)-->]Manalpha1 -4GlcNalpha1-6[acyl- >]inositol-PO(4)-lipid and Manalpha1-2Manalpha1-2Manalpha1-6Manalpha1-4G lcNalpha1-6[acyl-->]inos itol-PO(4)-lipid. Subcellular localization studies of a tagged version of YLL031c suggest that this protein is mainly in the ER, in contrast to Gpi7p, which is found at the cell surface. The data are compatible with the idea that YLL031c transfers the ethanolaminephosphate to the inner alpha1-2-linked mannose, i.e. the group that links the GPI lipid anchor to proteins, whereas Mcd4p and Gpi7p transfer ethanolaminephosphate onto the alpha1 4- and alpha1-6-linked mannoses of the GPI anchor, respectively. PMID- 10823838 TI - Identification of structural elements important for matrix metalloproteinase type V collagenolytic activity as revealed by chimeric enzymes. Role of fibronectin like domain and active site of gelatinase B. AB - Digestion of type V collagen by the gelatinases is an important step in tumor cell metastasis because this collagen maintains the integrity of the extracellular matrix that must be breached during this pathological process. However, the structural elements that provide the gelatinases with this unique proteolytic activity among matrix metalloproteinases had not been thoroughly defined. To identify these elements, we examined the substrate specificity of chimeric enzymes containing domains of gelatinase B and fibroblast collagenase. We have found that the addition of the fibronectin-like domain of gelatinase B to fibroblast collagenase is sufficient to endow the enzyme with the ability to cleave type V collagen. In addition, the substitution of the catalytic zinc binding active site region of fibroblast collagenase with that of gelatinase B increased the catalytic efficiency of the enzyme 3- to 4-fold. This observation led to the identification of amino acid residues, Leu(397), Ala(406), Asp(410), and Pro(415), in this region of gelatinase B that are important for its efficient catalysis as determined by substituting these amino acids with the corresponding residues from fibroblast collagenase. Leu(397) and Ala(406) are important for the general proteolytic activity of the enzyme, whereas Asp(410) and Pro(415) specifically enhance its ability to cleave type V collagen and gelatin, respectively. These data provide fundamental information about the structural elements that distinguish the gelatinases from other matrix metalloproteinases in terms of substrate specificity and catalytic efficiency. PMID- 10823839 TI - Mediation by the protein-tyrosine kinase Tec of signaling between the B cell antigen receptor and Dok-1. AB - A variety of growth factor receptors induce the tyrosine phosphorylation of a nonreceptor protein-tyrosine kinase Tec as well as that of a Tec-binding protein of 62 kDa. Given the similarity in properties between this 62-kDa protein and p62(Dok-1), the possibility that these two proteins are identical was investigated. Overexpression of a constitutively active form of Tec in a pro-B cell line induced the hyperphosphorylation of endogenous Dok-1. Tec also associated with Dok-1 in a phosphorylation-dependent manner in 293 cells. Tec mediated marked phosphorylation of Dok-1 both in vivo and in vitro, and this effect required both the Tec homology and Src homology 2 domains of Tec in addition to its kinase activity. Expression of Dok-1 in 293 cells induced inhibition of Ras activity, suggesting that Dok-1 is a negative regulator of Ras. In the immature B cell line Ramos, cross-linking of the B cell antigen receptor (BCR) resulted in tyrosine phosphorylation of Dok-1, and this effect was markedly inhibited by expression of dominant negative mutants of Tec. Furthermore, overexpression of Dok-1 inhibited activation of the c-fos promoter induced by stimulation of the BCR. These results suggest that Tec is an important mediator of signaling from the BCR to Dok-1. PMID- 10823840 TI - Tumor necrosis factor-alpha activation of NF-kappa B requires the phosphorylation of Ser-471 in the transactivation domain of c-Rel. AB - Activation of the transcription factor NF-kappaB is controlled at two levels in resting T cells: an initial activation induced by the triggering of the TcR.CD3 complex and a second phase controlled by paracrine- or autocrine-secreted TNFalpha. The initial phase is regulated by p65 (RelA), whereas the second one is mainly dependent on c-Rel. We describe here a mutant clone, D6, derived from the parental T lymphoblastic line Jurkat that fails to activate NF-kappaB upon TNFalpha stimulation. This clone had no alteration in tumor necrosis factor-alpha (TNFalpha) signaling pathways nor in IkappaBalpha, -beta, or -epsilon expression and degradation. However, TNFalpha induced an exacerbated apoptotic response in this clone compared with Jurkat cells. This mutant clone showed a defect in the intermediate-late translocation of c-Rel to the nucleus promoted by TNFalpha stimulation, whereas early translocation is not affected. Activation or translocation of p65-containing complexes was not altered in this mutant clone. Sequencing of the c-Rel gene from this clone revealed a mutation of Ser-471 to Asn in the transactivation domain. The mutant S471N transactivation domain fused to the Gal4 DNA binding domain could not be activated by TNFalpha, unlike the wild type. Moreover, the overexpression of the mutant protein c-Rel S471N into Jurkat cells abolished TNFalpha-induced NF-kappaB activity, thus demonstrating that this mutation is responsible for the failure of TNFalpha stimulation of NF kappaB. Moreover, extracts from TNFalpha-stimulated Jurkat cells phosphorylated in vitro recombinant wild type GST-c-Rel 464-481 but not the GST-c-Rel mutant. Thus, TNFalpha-induced phosphorylation of Ser-471 seems to be absolutely necessary for TNFalpha activation of c-Rel. PMID- 10823841 TI - Constitutive activation of STAT5 by a point mutation in the SH2 domain. AB - We previously identified a constitutively active form of STAT (signal transducer and activator of transcription) 5A by polymerase chain reaction-driven random mutagenesis followed by retrovirus-mediated expression screening, which had two point mutations in the DNA-binding and transcriptional activation domains, and was designated STAT5A1*6. STAT5A1*6 showed markedly elevated DNA binding and transactivation activities with stable tyrosine phosphorylation and nuclear accumulation, and conferred autonomous cell growth on interleukin 3-dependent Ba/F3 cells. We now report another constitutively active mutant, STAT5A-N642H which has a single point mutation (N642H) in its SH2 domain, identified using the same strategy as that used to identify STAT5A1*6. STAT5A-N642H showed identical properties to those of STAT5A1*6 both biochemically and biologically. Interestingly the mutation in STAT5A-N642H resulted in restoration of the conserved critical histidine which is involved in the binding of phosphotyrosine in the majority of SH2-containing proteins. Introduction of an additional mutation (Y694F) to STAT5A-N642H, which disrupted critical tyrosine 694 required for dimerization of STAT5, abolished all the activities manifested by the mutant STAT5A-N642H, which indicates that dimerization is required for the activity of STAT5A-N642H as was the case for the wild-type STAT5A. The present findings also show that different mutations rendered STAT5A constitutively active, through a common mechanism, which is similar to that of physiological activation. PMID- 10823842 TI - Differential display identification of 40 genes with altered expression in activated human smooth muscle cells. Local expression in atherosclerotic lesions of smags, smooth muscle activation-specific genes. AB - Detailed knowledge on the molecular and cellular mechanisms that control (re) differentiation of vascular smooth muscle cells (SMCs) is critical to understanding the pathological processes underlying atherogenesis. We identified by differential display/reverse transcriptase-polymerase chain reaction 40 genes with altered expression in cultured SMCs upon stimulation with the conditioned medium of activated macrophages. This set of genes comprises 10 known genes and 30 novel genes, which we call "smags" (for smooth muscle activation-specific genes). To determine the in vivo significance of these (novel) genes in atherogenesis, we performed in situ hybridization experiments on vascular tissue. Specifically, FLICE (Fas-associated death domain-like interleukin-1beta converting enzyme)-like inhibitory protein (FLIP) is expressed in neointimal SMCs as well as in lesion macrophages and endothelial cells, whereas the expression of the novel genes smag-63, smag-64, and smag-84 is restricted to neointimal SMCs. Characterization of full-length smag-64 cDNA revealed that it encodes a novel protein of 66 amino acids. smag-82 cDNA comprises the complete, unknown, 3' untranslated region of fibroblast growth factor-5. Collectively, our results illustrate the complex changes of SMC gene expression that occur in response to stimulation with cytokines and growth factors secreted by activated macrophages. Moreover, we identified interesting candidate genes that may play a role in the differentiation of SMCs during atherogenesis. PMID- 10823843 TI - Efficient particle production by minimal Gag constructs which retain the carboxy terminal domain of human immunodeficiency virus type 1 capsid-p2 and a late assembly domain. AB - The human immunodeficiency virus type 1 (HIV-1) Gag precursor Pr55(gag) by itself is capable of assembling into retrovirus-like particles (VLP). In the present study, we attempted to identify the minimal Gag sequences required for the formation of VLP. Our results show that about 80% of Pr55(gag) can be either deleted or replaced by heterologous sequences without significantly compromising VLP production. The smallest chimeric molecule still able to efficiently form VLP was only about 16 kDa. This minimal Gag construct contained the leucine zipper domain of the yeast transcription factor GCN4 to substitute for the assembly function of nucleocapsid (NC), followed by a P-P-P-P-Y motif to provide late budding (L) domain function, and retained only the myristylation signal and the C terminal capsid-p2 domain of Pr55(gag). We also show that the L domain function of HIV-1 p6(gag) is not dependent on the presence of an active viral protease and that the NC domain of Pr55(gag) is dispensable for the incorporation of Vpr into VLP. PMID- 10823844 TI - Human immunodeficiency virus (HIV)-positive sera obtained shortly after seroconversion neutralize autologous HIV type 1 isolates on primary macrophages but not on lymphocytes. AB - The aim of this study was to analyze the role of humoral immunity in early human immunodeficiency virus (HIV) infection. As neutralizing activities in HIV positive sera are rarely detectable earlier than 9 to 12 months after infection using primary lymphocytes as target cells in neutralization assays, humoral immunity is generally thought not to contribute significantly to early virus control in the patients. Besides lymphocytes, cells of the monocyte/macrophage lineage are known to be important target cells for HIV in vivo during the establishment of the infection. Therefore, we studied the neutralization of early primary HIV isolates by autologous serum samples using primary macrophages as target cells in the neutralization assays. We analyzed neutralizing activities against the autologous HIV-1 isolates in 10 patients' sera taken shortly after seroconversion, both on primary macrophages and, for comparison, on lymphocytes. Viruses were isolated and expanded in primary mixed cultures containing macrophages and lymphocytes in order to avoid selection for one particular cell type. All viruses replicated to different degrees in macrophages and lymphocytes; nine had a nonsyncytium-inducing phenotype, and one was syncytium inducing. The detection of neutralizing antibodies in acute primary HIV infection depended on the target cells used. Confirming previous studies, we did not find neutralizing activities on lymphocytes at this early time point. In contrast, neutralizing activities were detectable in the same sera if primary macrophages were used as target cells. Differences in neutralizing activities on macrophages and lymphocytes were not due to different virus variants being present in the different cell systems, as gp120 sequences derived from both cell types were homogeneous. Neutralization activities on macrophages did not correlate with the amount of beta-chemokines in the sera. As affinity-purified immunoglobulin G preparations from an early patient serum also exhibited neutralization of the autologous virus isolate on primary macrophages, but not on lymphocytes, neutralization is very likely due to antibodies against viral epitopes necessary for infection of macrophages but not for infection of lymphocytes. Our data suggest that, along with cell-mediated immunity, humoral immunity may contribute to the reduction of primary viremia in the patient. This was further supported by a certain association between neutralizing antibody titers on macrophages and viral load in the patients. PMID- 10823845 TI - Rubella virus nonstructural protein protease domains involved in trans- and cis cleavage activities. AB - Rubella virus (RV) genomic RNA contains two large open reading frames (ORFs): a 5'-proximal ORF encoding nonstructural proteins (NSPs) that function primarily in viral RNA replication and a 3'-proximal ORF encoding the viral structural proteins. Proteolytic processing of the RV NSP ORF translation product p200 is essential for viral replication. Processing of p200 to two mature products (p150 and p90) in the order NH(2)-p150-p90-COOH is carried out by an RV-encoded protease residing in the C-terminal region of p150. The RV nonstructural protease (NS-pro) belongs to a viral papain-like protease family that cleaves the polyprotein both in trans and in cis. A conserved X domain of unknown function was found from previous sequence analysis to be associated with NS-pro. To define the domains responsible for cis- and trans-cleavage activities and the function of the X domain in terms of protease activity, an in vitro translation system was employed. We demonstrated that the NSP region from residue 920 to 1296 is necessary for trans-cleavage activity. The domain from residue 920 to 1020 is not required for cis-cleavage activity. The X domain located between residues 834 and 940, outside the regions responsible for both cis- and trans-cleavage activities of NS-pro, was found to be important for NS-pro trans-cleavage activity but not for cis-cleavage activity. Analysis of sequence homology and secondary structure of the RV NS-pro catalytic region reveals a folding structure similar to that of papain. PMID- 10823846 TI - Efficient DNA transfection mediated by the C-terminal domain of human immunodeficiency virus type 1 viral protein R. AB - Viral protein R (Vpr) of human immunodeficiency virus type 1 is produced late in the virus life cycle and is assembled into the virion through binding to the Gag protein. It is known to play a significant role early in the viral life cycle by facilitating the nuclear import of the preintegration complex in nondividing cells. Vpr is also able to interact with nucleic acids, and we show here that it induces condensation of plasmid DNA. We have explored the possibility of using these properties in DNA transfection experiments. We report that the C-terminal half of the protein (Vpr(52-96)) mediates DNA transfection in a variety of human and nonhuman cell lines with efficiencies comparable to those of the best-known transfection agents. Compared with polylysine, a standard polycationic transfection reagent, Vpr(52-96) was 10- to 1,000-fold more active. Vpr(52-96) DNA complexes were able to reach the cell nucleus through a pH-independent mechanism. These observations possibly identify an alternate pathway for DNA transfection. PMID- 10823847 TI - Opposite effects of dextran sulfate 500, the polyene antibiotic MS-8209, and Congo red on accumulation of the protease-resistant isoform of PrP in the spleens of mice inoculated intraperitoneally with the scrapie agent. AB - The mode and the site of action of the major antiscrapie drugs have been studied by investigating their effects on the abnormal protease-resistant isoform of PrP (PrPres) and on its accumulation in mouse spleen. Day-by-day PrPres accumulation in the spleen and in other peripheral organs was first monitored to describe the early steps of scrapie pathogenesis. Three phases were identified: the detection of scrapie inoculum on the day of scrapie infection, a clearance phase, and then the peripheral accumulation of PrPres. In a second step, the effects of the polyene antibiotic MS-8209, the polyanion dextran sulfate 500 (DS500), and Congo red were assessed on these phases, after the drugs were coincubated with scrapie inoculum. Highly different mechanisms and sites of action were apparent. MS-8209 had a weak effect on the accumulation of PrPres in spleen, suggesting another site of intervention for this drug. DS500 delayed the beginning of the clearance phase but then blocked PrPres synthesis for a long period of time, probably because of its immunological effects on the spleen. Surprisingly, Congo red suppressed the clearance phase of scrapie inoculum and then increased transiently accumulation of PrPres in spleen. We showed in vitro that this effect was related to a direct enhancement of the protease resistance of PrPres by the drug. PMID- 10823848 TI - The double-stranded RNA-binding protein Staufen is incorporated in human immunodeficiency virus type 1: evidence for a role in genomic RNA encapsidation. AB - Human Staufen (hStau), a double-stranded RNA (dsRNA)-binding protein that is involved in mRNA transport, is incorporated in human immunodeficiency virus type 1 (HIV-1) and in other retroviruses, including HIV-2 and Moloney murine leukemia virus. Sucrose and Optiprep gradient analyses reveal cosedimentation of hStau with purified HIV-1, while subtilisin assays demonstrate that it is internalized. hStau incorporation in HIV-1 is selective, is dependent on an intact functional dsRNA-binding domain, and quantitatively correlates with levels of encapsidated HIV-1 genomic RNA. By coimmunoprecipitation and reverse transcription-PCR analyses, we demonstrate that hStau is associated with HIV-1 genomic RNA in HIV-1 expressing cells and purified virus. Overexpression of hStau enhances virion incorporation levels, and a corresponding, threefold increase in HIV-1 genomic RNA encapsidation levels. This coordinated increase in hStau and genomic RNA packaging had a significant negative effect on viral infectivity. This study is the first to describe hStau within HIV-1 particles and provides evidence that hStau binds HIV-1 genomic RNA, indicating that it may be implicated in retroviral genome selection and packaging into assembling virions. PMID- 10823850 TI - Dose-dependent changes in influenza virus-infected dendritic cells result in increased allogeneic T-cell proliferation at low, but not high, doses of virus. AB - During the acute phase of infection with influenza A virus, the degree of lymphopenia correlates with severity of disease. Factors that contribute to T cell activation during influenza virus infection may contribute to this observation. Since the immune response is initiated when dendritic cells (DC) interact with T cells, we have established an in vitro system to examine the effects of influenza virus infection on DC function. Our results show that allogeneic T-cell proliferation was dependent on the dose of A/PR/8/34 used to infect DC, with enhanced responses at low, but not high, multiplicities of infection. The lack of enhancement at high virus doses was not primarily due to the increased rate of DC apoptosis, but required viral replication and neuraminidase (NA) activity. Clusters that formed between DC or between DC and T cells were also dependent on the viral dose. This change in cellular interaction may oppose T-cell proliferation in response to DC infected with high doses of PR8, since the increased contact between DC resulted in the exclusion of T cells. The enhanced alloreactive T-cell response was restored by neutralization of transforming growth factor beta1 (TGF-beta1). It is likely that NA present on viral particles released from DC infected with high doses of PR8 activates TGF beta1. Future studies will determine the mechanism by which TGF-beta1 modifies the in vitro T-cell response and address the contribution of this cytokine to the lymphopenia observed in severe disease. PMID- 10823849 TI - Polymorphism in the interleukin-4 promoter affects acquisition of human immunodeficiency virus type 1 syncytium-inducing phenotype. AB - The emergence of syncytium-inducing (SI) variants of human immunodeficiency virus type 1 (HIV-1) in infected individuals is an indicator of poor prognosis and is often correlated with faster CD4(+) cell depletion and rapid disease progression. Interleukin-4 (IL-4) is a pleiotropic cytokine with various immune-modulating functions including induction of immunoglobulin E (IgE) production in B cells, down-regulation of CCR5 (a coreceptor for HIV-1 non-SI [NSI] strains), and up regulation of CXCR4 (a coreceptor for HIV-1 SI variants). Here we show that homozygosity of a polymorphism in the IL-4 promoter region, IL-4 -589T, is correlated with increased rates of SI variant acquisition in HIV-1-infected individuals in Japan. This mutation was also shown to be associated with elevated serum IgE levels in HIV-1-infected individuals, especially in those at advanced stages of disease. In contrast, neither a triallele polymorphism in IL-10, another Th2 cytokine, nor a biallele polymorphism in the RANTES promoter affected acquisition of the SI phenotype. This finding suggested that IL-4-589T increases IL-4 production in the human body and thus accelerates the phenotypic switch of HIV-1 from NSI to SI and possibly disease progression of AIDS. PMID- 10823851 TI - H-2D(b-/-) mice are susceptible to persistent infection by Theiler's virus. AB - H-2(b) mice are resistant to persistent infection of the central nervous system by Theiler's virus. They clear the infection 7 to 10 days after intracranial inoculation. Resistance maps to the H-2D gene and not to the H-2K gene and is associated with a potent antiviral cytotoxic T-lymphocyte (CTL) response. We used H-2(b) mice in which the H-2D or the H-2K gene had been inactivated to dissect the respective roles of these genes in resistance. We report that H-2D(-/-) but not H-2K(-/-) mice were susceptible to persistent infection. Furthermore, whereas H-2K(-/-) mice mounted a vigorous virus-specific CTL response, similar to that of control C57BL/6 mice, the CTL response of H-2D(-/-) mice was nil or minimal. Using target cells transfected with the H-2D(b) or the H-2K(b) gene, we showed that the H-2K-restricted CTL response against the virus was minimal in H-2D(-/-) mice. These results demonstrate that the H-2D(b) and H-2K(b) genes play nonredundant roles in the resistance to this persistent infection. PMID- 10823852 TI - Recombinant chimeric yellow fever-dengue type 2 virus is immunogenic and protective in nonhuman primates. AB - A chimeric yellow fever (YF)-dengue type 2 (dengue-2) virus (ChimeriVax-D2) was constructed using a recombinant cDNA infectious clone of a YF vaccine strain (YF 17D) as a backbone into which we inserted the premembrane (prM) and envelope (E) genes of dengue-2 virus (strain PUO-218 from a case of dengue fever in Bangkok, Thailand). The chimeric virus was recovered from the supernatant of Vero cells transfected with RNA transcripts and amplified once in these cells to yield a titer of 6.3 log(10) PFU/ml. The ChimeriVax-D2 was not neurovirulent for 4-week old outbred mice inoculated intracerebrally. This virus was evaluated in rhesus monkeys for its safety (induction of viremia) and protective efficacy (induction of anti-dengue-2 neutralizing antibodies and protection against challenge). In one experiment, groups of non-YF-immune monkeys received graded doses of ChimeriVax-D2; a control group received only the vaccine diluents. All monkeys (except the control group) developed a brief viremia and showed no signs of illness. Sixty-two days postimmunization, animals were challenged with 5.0 log(10) focus forming units (FFU) of a wild-type dengue-2 virus. No viremia (<1.7 log(10) FFU/ml) was detected in any vaccinated group, whereas all animals in the placebo control group developed viremia. All vaccinated monkeys developed neutralizing antibodies in a dose-dependent response. In another experiment, viremia and production of neutralizing antibodies were determined in YF-immune monkeys that received either ChimeriVax-D2 or a wild-type dengue-2 virus. Low viremia was detected in ChimeriVax-D2-inoculated monkeys, whereas all dengue-2 immunized animals became viremic. All of these animals were protected against challenge with a wild-type dengue-2 virus, whereas all YF-immune monkeys and nonimmune controls became viremic upon challenge. Genetic stability of ChimeriVax D2 was assessed by continuous in vitro passage in VeroPM cells. The titer of ChimeriVax-D2, the attenuated phenotype for 4-week-old mice, and the sequence of the inserted prME genes were unchanged after 18 passages in Vero cells. The high replication efficiency, attenuation phenotype in mice and monkeys, immunogenicity and protective efficacy, and genomic stability of ChimeriVax-D2 justify it as a novel vaccine candidate to be evaluated in humans. PMID- 10823853 TI - A yeast-based genetic system for functional analysis of viral mRNA capping enzymes. AB - Virus-encoded mRNA capping enzymes are attractive targets for antiviral therapy, but functional studies have been limited by the lack of genetically tractable in vivo systems that focus exclusively on the RNA-processing activities of the viral proteins. Here we have developed such a system by engineering a viral capping enzyme-vaccinia virus D1(1-545)p, an RNA triphosphatase and RNA guanylyltransferase-to function in the budding yeast Saccharomyces cerevisiae in lieu of the endogenous fungal triphosphatase (Cet1p) and guanylyltransferase (Ceg1p). This was accomplished by fusion of D1(1-545)p to the C-terminal guanylyltransferase domain of mammalian capping enzyme, Mce1(211-597)p, which serves as a vehicle to target the viral capping enzyme to the RNA polymerase II elongation complex. An inactivating mutation (K294A) of the mammalian guanylyltransferase active site in the fusion protein had no impact on genetic complementation of cet1Deltaceg1Delta cells, thus proving that (i) the viral guanylyltransferase was active in vivo and (ii) the mammalian domain can serve purely as a chaperone to direct other proteins to the transcription complex. Alanine scanning had identified five amino acids of vaccinia virus capping enzyme Glu37, Glu39, Arg77, Glu192, and Glu194-that are essential for gamma phosphate cleavage in vitro. Here we show that the introduction of mutation E37A, R77A, or E192A into the fusion protein abrogates RNA triphosphatase function in vivo. The essential residues are located within three motifs that define a family of viral and fungal metal-dependent phosphohydrolases with a distinctive capacity to hydrolyze nucleoside triphosphates to nucleoside diphosphates in the presence of manganese or cobalt. The acidic residues Glu37, Glu39, and Glu192 likely comprise the metal-binding site of vaccinia virus triphosphatase, insofar as their replacement by glutamine abolishes the RNA triphosphatase and ATPase activities. PMID- 10823854 TI - Gamma interferon is not required for mucosal cytotoxic T-lymphocyte responses or heterosubtypic immunity to influenza A virus infection in mice. AB - Heterosubtypic immunity (HSI) is defined as cross-protection against influenza virus of a different serotype than the virus initially encountered and is thought to be mediated by influenza virus-specific cytotoxic T lymphocytes (CTL). Since gamma interferon (IFN-gamma) stimulates cytotoxic cells, including antigen specific CTL which may control virus replication by secretion of antiviral cytokines such as tumor necrosis factor alpha and IFN-gamma, we have investigated the mechanism of HSI by analyzing the role of IFN-gamma for HSI in IFN-gamma gene deleted (IFN-gamma(-/-)) mice. It has been reported that IFN-gamma is not required for recovery from primary infection with influenza virus but is important for HSI. Here, we conclusively show that IFN-gamma is not required for induction of secondary influenza virus-specific CTL responses in mediastinal lymph nodes and HSI to lethal influenza A virus infection. Although T helper 2 (Th2)-type cytokines were upregulated in the lungs of IFN-gamma(-/-) mice after virus challenge, either Th1- or Th2-biased responses could provide heterosubtypic protection. Furthermore, titers of serum-neutralizing and cross-reactive antibodies to conserved nucleoprotein in IFN-gamma(-/-) mice did not differ significantly from those in immunocompetent mice. These results indicate that lack of IFN-gamma does not impair cross-reactive virus-specific immune responses and HSI to lethal infection with influenza virus. Our findings provide new insight for the mechanisms of HSI and should be valuable in the development of protective mucosal vaccines against variant virus strains, such as influenza and human immunodeficiency virus. PMID- 10823855 TI - Evidence that the 5'-end cap structure is essential for encapsidation of hepatitis B virus pregenomic RNA. AB - Hepatitis B virus (HBV) replicates by reverse transcription of an RNA intermediate, the pregenomic RNA. The first step of HBV genome replication is the encapsidation of the pregenomic RNA encoding the encapsidation signal, termed epsilon, into the core particles, which is preceded by recognition and binding of HBV DNA polymerase to epsilon. The pregenomic RNA contains two identical epsilon elements due to its terminal redundancy: one near the 5' end and another near the 3' end. Despite the fact that both epsilon elements have an identical sequence, only the 5' epsilon, but not the 3' epsilon, is functional for encapsidation. To understand the molecular nature of this position effect, we made a series of lacZ RNA expression plasmids which contain the epsilon element at various positions from the 5' end of the transcripts. Following transfection, the lacZ RNAs in cytoplasmic core particles were measured by RNase protection assay for encapsidation. The results indicated that the lacZ RNAs with epsilon positioned up to 65 nucleotides from the 5' end were encapsidated, whereas the lacZ RNAs with epsilon positioned further downstream were not. Interestingly, the cap-free lacZ RNA transcribed by T7 RNA polymerase was not encapsidated, implying that the 5' cap structure is required for encapsidation of the pregenomic RNA. We hypothesized that HBV DNA polymerase must somehow recognize the cap structure and/or its associated factors, as well as the 5' epsilon, for encapsidation to occur. PMID- 10823856 TI - Interactions between hepatitis delta virus proteins. AB - The 195- and 214-amino-acid (aa) forms of the delta protein (deltaAg-S and deltaAg-L, respectively) of hepatitis delta virus (HDV) differ only in the 19-aa C-terminal extension unique to deltaAg-L. deltaAg-S is needed for genome replication, while deltaAg-L is needed for particle assembly. These proteins share a region at aa 12 to 60, which mediates protein-protein interactions essential for HDV replication. H. Zuccola et al. (Structure 6:821-830, 1998) reported a crystal structure for a peptide spanning this region which demonstrates an antiparallel coiled-coil dimer interaction with the potential to form tetramers of dimers. Our studies tested whether predictions based on this structure could be extrapolated to conditions where the peptide was replaced by full-length deltaAg-S or deltaAg-L, and when the assays were not in vitro but in vivo. Nine amino acids that are conserved between several isolates of HDV and predicted to be important in multimerization were mutated to alanine on both deltaAg-S and deltaAg-L. We found that the predicted hierarchy of importance of these nine mutations correlated to a significant extent with the observed in vivo effects on the ability of these proteins to (i) support in trans the replication of the HDV genome when expressed on deltaAg-S and (ii) act as dominant-negative inhibitors of replication when expressed on deltaAg-L. We thus infer that these biological activities of deltaAg depend on ordered protein-protein interactions. PMID- 10823857 TI - Reovirus nonstructural protein muNS binds to core particles but does not inhibit their transcription and capping activities. AB - Previous studies provided evidence that nonstructural protein muNS of mammalian reoviruses is present in particle assembly intermediates isolated from infected cells. Morgan and Zweerink (Virology 68:455-466, 1975) showed that a subset of these intermediates, which can synthesize the viral plus strand RNA transcripts in vitro, comprise core-like particles plus large amounts of muNS. Given the possible role of muNS in particle assembly and/or transcription implied by those findings, we tested whether recombinant muNS can bind to cores in vitro. The muNS protein bound to cores, but not to two particle forms, virions and intermediate subvirion particles, that contain additional outer-capsid proteins. Incubating cores with increasing amounts of muNS resulted in particle complexes of progressively decreasing buoyant density, approaching the density of protein alone when very large amounts of muNS were bound. Thus, the muNS-core interaction did not exhibit saturation or a defined stoichiometry. Negative-stain electron microscopy of the muNS-bound cores revealed that the cores were intact and linked together in large complexes by an amorphous density, which we ascribe to muNS. The muNS-core complexes retained the capacity to synthesize the viral plus strand transcripts as well as the capacity to add methylated caps to the 5' ends of the transcripts. In vitro competition assays showed that mixing muNS with cores greatly reduced the formation of recoated cores by stoichiometric binding of outer-capsid proteins mu1 and sigma3. These findings are consistent with the presence of muNS in transcriptase particles as described previously and suggest that, by binding to cores in the infected cell, muNS may block or delay outer capsid assembly and allow continued transcription by these particles. PMID- 10823858 TI - Endotoxin stimulates liver macrophages to release mediators that inhibit an early step in hepadnavirus replication. AB - Hepadnaviruses are known to be sensitive to various extracellular mediators. Therefore, bacterial endotoxin, which induces the secretion of proinflammatory mediators in the liver, was studied for its effect on hepadnavirus infection in vitro using the duck hepatitis B virus (DHBV) model. In initial experiments, endotoxin was shown to inhibit DHBV replication in primary duck hepatocyte cultures prepared by standard collagenase perfusion. As a primary endotoxin target, hepatic nonparenchymal cells (NPC) contaminating primary hepatocyte cultures, and among these probably macrophages (Kupffer cells), were identified to secrete polypeptide mediators into the cell culture medium. When added during DHBV infection, these mediators elicited the principal antiviral effect in a dose dependent fashion. On the molecular level, they inhibited accumulation of viral proteins as well as amplification of the nuclear extrachromosomal DHBV DNA templates. In hepatocytes with an established DHBV infection, DHBV protein and progeny virus production was inhibited while the levels of established nuclear DHBV DNA templates and viral transcripts remained unaffected. Finally, in hepatocytes infected with a replication-deficient recombinant DHBV-green fluorescent protein (GFP) virus, the endotoxin-induced mediators markedly reduced GFP expression from chimeric DHBV-GFP transcripts, indicating that the major effect is at a level of translation of viral RNAs. Taken together, the data obtained demonstrate that antiviral mediators, and among these the cytokines alpha interferon (IFN-alpha) and IFN-gamma, are released from hepatic NPC, most probably liver macrophages, upon endotoxin stimulation; furthermore, these mediators act at a posttranscriptional step of hepadnavirus replication. PMID- 10823859 TI - Competing death programs in poliovirus-infected cells: commitment switch in the middle of the infectious cycle. AB - Productive poliovirus infection of HeLa cells leads to the canonical cytopathic effect (CPE), whereas certain types of abortive infection result in apoptosis. To define the time course of commitment to the different types of poliovirus-induced death, inhibitors of viral replication (guanidine HCl) or translation (cycloheximide) were added at different times postinfection (p.i.). Early in the infection (during the first approximately 2 h p.i.), predominantly proapoptotic viral function was expressed, rendering the cells committed to apoptosis, which developed several hours after viral expression was arrested. In the middle of infection, concomitantly with the onset of fast generation of viral progeny, the implementation of the viral apoptotic program was abruptly interrupted. In particular, activation of an Asp-Glu-Val-Asp (DEVD)-specific caspase(s) occurring in the apoptosis-committed cells was prevented by the ongoing productive infection. Simultaneously, the cells retaining normal or nearly normal morphology became committed to CPE, which eventually developed regardless of whether or not further viral expression was allowed to proceed. The implementation of the poliovirus-induced apoptotic program was suppressed in HeLa cells overexpressing the Bcl-2 protein, indicating that the fate of poliovirus-infected cells depends on the balance of host and viral pro- and antiapoptotic factors. PMID- 10823860 TI - Adaptation and selection of prion protein strain conformations following interspecies transmission of transmissible mink encephalopathy. AB - Interspecies transmission of the transmissible spongiform encephalopathies (TSEs), or prion diseases, can result in the adaptation and selection of TSE strains with an expanded host range and increased virulence such as in the case of bovine spongiform encephalopathy and variant Creutzfeldt-Jakob disease. To investigate TSE strain adaptation, we serially passaged a biological clone of transmissible mink encephalopathy (TME) into Syrian golden hamsters and examined the selection of distinct strain phenotypes and conformations of the disease specific isoform of the prion protein (PrP(Sc)). The long-incubation-period drowsy (DY) TME strain was the predominate strain, based on the presence of its strain-specific PrP(Sc) following interspecies passage. Additional serial passages in hamsters resulted in the selection of the hyper (HY) TME PrP(Sc) strain-dependent conformation and its short incubation period phenotype unless the passages were performed with a low-dose inoculum (e.g., 10(-5) dilution), in which case the DY TME clinical phenotype continued to predominate. For both TME strains, the PrP(Sc) strain pattern preceded stabilization of the TME strain phenotype. These findings demonstrate that interspecies transmission of a single cloned TSE strain resulted in adaptation of at least two strain-associated PrP(Sc) conformations that underwent selection until one type of PrP(Sc) conformation and strain phenotype became predominant. To examine TME strain selection in the absence of host adaptation, hamsters were coinfected with hamster-adapted HY and DY TME. DY TME was able to interfere with the selection of the short-incubation HY TME phenotype. Coinfection could result in the DY TME phenotype and PrP(Sc) conformation on first passage, but on subsequent passages, the disease pattern converted to HY TME. These findings indicate that during TSE strain adaptation, there is selection of a strain-specific PrP(Sc) conformation that can determine the TSE strain phenotype. PMID- 10823861 TI - Identification by phage display and characterization of two neutralizing chimpanzee monoclonal antibodies to the hepatitis E virus capsid protein. AB - Two monoclonal antibodies (MAbs) against the ORF2 protein of the SAR-55 strain of hepatitis E virus (HEV) were isolated by phage display from a cDNA library of chimpanzee (Pan troglodytes) gamma1/kappa antibody genes. Both MAbs, HEV#4 and HEV#31, bound to reduced, denatured open reading frame 2 (ORF2) protein in a Western blot, suggesting that they recognize linear epitopes. The affinities (equilibrium dissociation constants, K(d)) for the SAR-55 ORF2 protein were 1.7 nM for HEV#4 and 5.4 nM for HEV#31. The two MAbs also reacted in an enzyme-linked immunosorbent assay with recombinant ORF2 protein from a heterologous HEV, the Meng strain. Each MAb blocked the subsequent binding of the other MAb to homologous ORF2 protein in indirect competition assays, suggesting that they recognize the same or overlapping epitopes. Radioimmunoprecipitation assays suggested that at least part of the linear epitope(s) recognized by the two MAbs is located between amino acids 578 and 607. MAbs were mixed with homologous HEV in vitro and then inoculated into rhesus monkeys (Macaca mulatta) to determine their neutralizing ability. Whereas all control animals developed hepatitis (elevated liver enzyme levels in serum) and seroconverted to HEV, those receiving an inoculum incubated with either HEV#4 or HEV#31 were not infected. Therefore, each MAb neutralized the SAR-55 strain of HEV in vitro. PMID- 10823862 TI - Characterization of influenza virus NS1 protein by using a novel helper-virus free reverse genetic system. AB - We have developed a novel helper-virus-free reverse genetic system to genetically manipulate influenza A viruses. The RNPs, which were purified from the influenza A/WSN/33 (WSN) virus, were treated with RNase H in the presence of NS (nonstructural) cDNA fragments. This specifically digested the NS RNP. The NS digested RNPs thus obtained were transfected into cells together with the in vitro-reconstituted NS RNP. The NS-digested RNPs alone did not rescue viruses; however, cotransfection with the NS RNP did. This protocol was also used to rescue the NP transfectant. We obtained two NS1 mutants, dl12 and N110, using this protocol. The dl12 NS gene contains a deletion of 12 amino acids at positions 66 to 77 near the N terminus. This virus was temperature sensitive in Madin-Darby bovine kidney (MDBK) cells as well as in Vero cells. The translation of all viral proteins as well as cellular proteins was significantly disrupted during a later time of infection at the nonpermissive temperature of 39 degrees C. The N110 mutant consists of 110 amino acids which are the N-terminal 48% of the WSN virus NS1 protein. Growth of this virus was significantly reduced at any temperature. In the virus-infected cells, translation of the M1 protein was reduced to 10 to 20% of that of the wild-type virus; however, the translation of neither the nucleoprotein nor NS1 was significantly interfered with, indicating the important role of NS1 in translational stimulation of the M1 protein. PMID- 10823863 TI - Integrated hepatitis B virus DNA preserves the binding sequence of transcription factor Yin and Yang 1 at the virus-cell junction. AB - Accumulated findings have indicated that hepatitis B virus (HBV) DNA integrates into the cellular DNA of HBV-infected chronic hepatitis tissues. The integrated sequence (IS) of HBV DNA at the virus-cell junction is conserved in a 25-bp region which is adjacent to direct repeat 1. A cellular protein which we purified from the nuclear extract of HepG2 cells binds to the IS and was designated IS binding protein 3 (ISBP3). The amino acid sequence of ISBP3 was determined and found to be identical to that of transcription initiation factor Yin and Yang 1 (YY1). An antibody against C-terminal amino acids of YY1 recognized ISBP3 in a Western blot analysis and an electrophoretic mobility shift assay. Furthermore, ISBP3 also interacted with Y3, which corresponds to the YY1 binding sequence, to enhance intramolecular recombination of polyomavirus DNA. Although YY1 is known as a transcription factor, the IS did not exhibit any effect on the transcription of precore and pregenome RNAs. The possible involvement of YY1 in the intramolecular recombination of linear replicative HBV DNA has been examined (Y. Hayashi et al., unpublished data). Data suggest that YY1 is involved in the joining reaction between HBV DNA and cellular DNA to form the virus-cell junction. PMID- 10823864 TI - Rubella virus capsid associates with host cell protein p32 and localizes to mitochondria. AB - Togavirus nucleocapsids have a characteristic icosahedral structure and are composed of multiple copies of a capsid protein complexed with genomic RNA. The assembly of rubella virus nucleocapsids is unique among togaviruses in that the process occurs late in virus assembly and in association with intracellular membranes. The goal of this study was to identify host cell proteins which may be involved in regulating rubella virus nucleocapsid assembly through their interactions with the capsid protein. Capsid was used as bait to screen a CV1 cDNA library using the yeast two-hybrid system. One protein that interacted strongly with capsid was p32, a cellular protein which is known to interact with other viral proteins. The interaction between capsid and p32 was confirmed using a number of different in vitro and in vivo methods, and the site of interaction between these two proteins was shown to be at the mitochondria. Interestingly, overexpression of the rubella virus structural proteins resulted in clustering of the mitochondria in the perinuclear region. The p32-binding site in capsid is a potentially phosphorylated region that overlaps the viral RNA-binding domain of capsid. Our results are consistent with the possibility that the interaction of p32 with capsid plays a role in the regulation of nucleocapsid assembly and/or virus-host interactions. PMID- 10823865 TI - Parameters of human immunodeficiency virus infection of human cervical tissue and inhibition by vaginal virucides. AB - Heterosexual transmission of human immunodeficiency virus (HIV) is the most frequent mode of infection worldwide. However, the immediate events between exposure to infectious virus and establishment of infection are still poorly understood. This study investigates parameters of HIV infection of human female genital tissue in vitro using an explant culture model. In particular, we investigated the role of the epithelium and virucidal agents in protection against HIV infection. We have demonstrated that the major target cells of infection reside below the genital epithelium, and thus HIV must cross this barrier to establish infection. Immune activation enhanced HIV infection of such subepithelial cells. Furthermore, our data suggest that genital epithelial cells were not susceptible to HIV infection, appear to play no part in the transfer of infectious virus across the epithelium, and thus may provide a barrier to infection. In addition, experiments using a panel of virucidal agents demonstrated differential efficiency to block HIV infection of subepithelial cells from partial to complete inhibition. This is the first demonstration that virucidal agents designed for topical vaginal use block HIV infection of genital tissue. Such agents have major implications for world health, as they will provide women with a mechanism of personal and covert protection from HIV infection. PMID- 10823866 TI - Expression of hepatitis C virus proteins interferes with the antiviral action of interferon independently of PKR-mediated control of protein synthesis. AB - Hepatitis C virus (HCV) of genotype 1 is the most resistant to interferon (IFN) therapy. Here, we have analyzed the response to IFN of the human cell line UHCV 11 engineered to inducibly express the entire HCV genotype 1a polyprotein. IFN treated, induced UHCV cells were found to better support the growth of encephalomyocarditis virus (EMCV) than IFN-treated, uninduced cells. This showed that expression of the HCV proteins allowed the development of a partial resistance to the antiviral action of IFN. The nonstructural 5A (NS5A) protein of HCV has been reported to inhibit PKR, an IFN-induced kinase involved in the antiviral action of IFN, at the level of control of protein synthesis through the phosphorylation of the initiation factor eIF2alpha (M. Gale, Jr., C. M. Blakely, B. Kwieciszewski, S. L. Tan, M. Dossett, N. M. Tang, M. J. Korth, S. J. Polyak, D. R. Gretch, and M. G. Katze, Mol. Cell. Biol. 18:5208-5218, 1998). Accordingly, cell lines inducibly expressing NS5A were found to rescue EMCV growth (S. J. Polyak, D. M. Paschal, S. McArdle, M. J. Gale, Jr., D. Moradpour, and D. R. Gretch, Hepatology 29:1262-1271, 1999). In the present study we analyzed whether the resistance of UHCV-11 cells to IFN could also be attributed to inhibition of PKR. Confocal laser scanning microscopy showed no colocalization of PKR, which is diffuse throughout the cytoplasm, and the induced HCV proteins, which localize around the nucleus within the endoplasmic reticulum. The effect of expression of HCV proteins on PKR activity was assayed in a reporter assay and by direct analysis of the in vivo phosphorylation of eIF2alpha after treatment of cells with poly(I)-poly(C). We found that neither PKR activity nor eIF2alpha phosphorylation was affected by coexpression of the HCV proteins. In conclusion, expression of HCV proteins in their biological context interferes with the development of the antiviral action of IFN. Although the possibility that some inhibition of PKR (by either NS5A or another viral protein) occurs at a very localized level cannot be excluded, the resistance to IFN, resulting from the expression of the HCV proteins, cannot be explained solely by inhibition of the negative control of translation by PKR. PMID- 10823867 TI - Primary murine small intestinal epithelial cells, maintained in long-term culture, are susceptible to rotavirus infection. AB - We describe a method for long-term culture of primary small intestinal epithelial cells (IEC) from suckling mice. IEC were digested from intestinal fragments as small intact units of epithelium (organoids) by using collagenase and dispase. IEC proliferated from organoids on a basement-membrane-coated culture surface and remained viable for 3 weeks. Cultured IEC had the morphologic and functional characteristics of immature enterocytes, notably sustained expression of cytokeratin and alkaline phosphatase. Few mesenchymal cells were present in the IEC cultures. IEC were also cultured from adult BALB/c mice and expressed major histocompatibility complex (MHC) class II antigens for at least 48 h in vitro. Primary IEC supported the growth of rhesus rotavirus (RRV) to a greater extent than a murine small intestinal cell line, m-IC(cl2). Cell-culture-adapted murine rotavirus strain EDIM infected primary IEC and m-IC(cl2) cells to a lesser extent than RRV. Wild-type EDIM did not infect either cell type. Long-term culture of primary murine small intestinal epithelial cells provides a method to study (i) virus-cell interactions, (ii) the capacity of IEC to act as antigen-presenting cells using a wide variety of MHC haplotypes, and (iii) IEC biology. PMID- 10823868 TI - Development and optimization of herpes simplex virus vectors for multiple long term gene delivery to the peripheral nervous system. AB - Herpes simplex virus (HSV) has often been suggested as a suitable vector for gene delivery to the peripheral nervous system as it naturally infects sensory nerve terminals before retrograde transport to the cell body in the spinal ganglia where latency is established. HSV vectors might therefore be particularly appropriate for the study and treatment of chronic pain following vector administration by relatively noninvasive peripheral routes. However parameters allowing safe and efficient gene delivery to spinal ganglia following peripheral vector inoculation, or the long-term expression of delivered genes, have not been comprehensively studied. We have identified combinations of deletions from the HSV genome which allow highly efficient gene delivery to spinal dorsal root ganglia (DRGs) following either footpad or sciatic nerve injection. These vectors have ICP34.5 deleted and have inactivating mutations in vmw65. We also report that peripheral replication is probably necessary for the efficient establishment of latency in vivo, as fully replication-incompetent HSV vectors allow efficient gene expression in DRGs only after peripheral inoculation at a high virus dose. Very low transduction efficiencies are otherwise achieved. In parallel, promoters have been developed that allow the long-term expression of individual or pairs of genes in DRGs by using elements from the latently active region of the virus to confer a long-term activity onto a number of promoters which otherwise function only in the short term. This work further defines elements and mechanisms within the latently active region that are necessary for long-term gene expression and for the first time allows multiple inserted genes to be expressed from HSV vectors during latency. PMID- 10823869 TI - Versatility of the accessory C proteins of Sendai virus: contribution to virus assembly as an additional role. AB - The P/C mRNA of Sendai virus (SeV) encodes a nested set of accessory proteins, C', C, Y1, and Y2, referred to collectively as C proteins, using the +1 frame relative to the open reading frame of phospho (P) protein and initiation codons at different positions. The C proteins appear to be basically nonstructural proteins as they are found abundantly in infected cells but greatly underrepresented in the virions. We previously created a 4C(-) SeV, which expresses none of the four C proteins, and concluded that the C proteins are categorically nonessential gene products but greatly contribute to viral full replication and infectivity (A. Kurotani et al., Genes Cells 3:111-124, 1998). Here, we further characterized the 4C(-) virus multiplication in cultured cells. The viral protein and mRNA synthesis was enhanced with the mutant virus relative to the parental wild-type (WT) SeV. However, the viral yields were greatly reduced. In addition, the 4C(-) virions appeared to be highly anomalous in size, shape, and sedimentation profile in a sucrose gradient and exhibited the ratios of infectivity to hemagglutination units significantly lower than those of the WT. In the WT infected cells, C proteins appeared to colocalize almost perfectly with the matrix (M) proteins, pretty well with an external envelope glycoprotein (hemagglutinin-neuraminidase [HN]), and very poorly with the internal P protein. In the absence of C proteins, there was a significant delay of the incorporation of M protein and both of the envelope proteins, HN and fusion (F) proteins, into progeny virions. These results strongly suggest that the accessory and basically nonstructural C proteins are critically required in the SeV assembly process. This role of C proteins was further found to be independent of their recently discovered function to counteract the antiviral action of interferon-alpha/beta. SeV C proteins thus appear to be quite versatile. PMID- 10823870 TI - Human cytomegalovirus replicates abortively in polymorphonuclear leukocytes after transfer from infected endothelial cells via transient microfusion events. AB - Using a recently developed model for in vitro generation of pp65-positive polymorphonuclear leukocytes (PMNLs), we demonstrated that PMNLs from immunocompetent subjects may harbor both infectious human cytomegalovirus (HCMV) and viral products (pp65, p72, DNA, and immediate-early [IE] and pp67 late mRNAs) as early as 60 min after coculture with human umbilical vein endothelial cells (HUVEC) or human embryonic lung fibroblasts (HELF) infected with a clinical HCMV isolate (VR6110) or other wild-type strains. The number of PMNLs positive for each viral parameter increased with coculture time. Using HELF infected with laboratory-adapted HCMV strains, only very small amounts of viral DNA and IE and late mRNAs were detected in PMNLs. A cellular mRNA, the vascular cell adhesion molecule-1 mRNA, which is abundantly present in both infected and uninfected HUVEC, was detected in much larger amounts in PMNLs cocultured with VR6110 infected cells than in controls. Coculture of PMNLs with VR6110-infected permissive cells in the presence or absence of RNA, protein, and viral DNA synthesis inhibitors showed that only IE genes were transcribed in PMNLs during coculture. Synthesis of IE transcripts in PMNLs was also supported by the finding that only the copy number of IE mRNA (and not the DNA or the pp67 mRNA) per infected PMNL increased markedly with time, and the pp67 to IE mRNA copy number ratio changed from greater than 10 in infected HUVEC to less than 1 in cocultured PMNLs. Fluorescent probe transfer experiments and electron microscopy studies indicated that transfer of infectious virus and viral products from infected cells to PMNLs is likely to be mediated by microfusion events induced by wild type strains only. In addition, HCMV pp65 and p72 were both shown to localize in the nucleus of the same PMNLs by double immunostaining. Two different mechanisms may explain the virus presence in PMNLs: (i) one major mechanism consists of transitory microfusion events (induced by wild-type strains only) of HUVEC or HELF and PMNLs with transfer of viable virus and biologically active viral material to PMNLs; and (ii) one minor mechanism, i.e., endocytosis, occurs with both wild-type and laboratory strains and leads to the acquisition of very small amounts of viral nucleic acids. In conclusion, HCMV replicates abortively in PMNLs, and wild-type strains and their products (as well as cellular metabolites and fluorescent dyes) are transferred to PMNLs, thus providing evidence for a potential mechanism of HCMV dissemination in vivo. PMID- 10823871 TI - The human immunodeficiency virus type 1 TAR RNA upper stem-loop plays distinct roles in reverse transcription and RNA packaging. AB - The human immunodeficiency virus type 1 (HIV-1) RNA genome is flanked by a repeated sequence (R) that is required for HIV-1 replication. The first 57 nucleotides of R form a stable stem-loop structure called the transactivation response element (TAR) that can interact with the virally encoded transcription activator protein, Tat, to promote high levels of gene expression. Recently, we demonstrated that TAR is also important for efficient HIV-1 reverse transcription, since HIV-1 mutated in the upper stem-loop of TAR showed a reduced ability both to initiate and to complete reverse transcription. We have analyzed a series of HIV-1 mutant viruses to better defined the structural or sequence elements required for natural endogenous reverse transcription and packaging of virion RNA. Our results indicate that the requirement for TAR in reverse transcription is conformation dependent, since mutants with mutations that alter the upper stem-loop orientation are defective for reverse transcription initiation and have minor defects in RNA packaging. In contrast, TAR mutations that allowed the formation of alternative upper stem-loop structure greatly reduced RNA packaging but did not affect reverse transcription efficiency. These results are consistent with direct involvement of the upper stem-loop structure in packaging of genomic RNA and suggest that the TAR RNA stem-loop from nucleotide +18 to +42 interacts with other components of the reverse transcription initiation complex to promote efficient reverse transcription. PMID- 10823872 TI - Mouse hepatitis virus replicase proteins associate with two distinct populations of intracellular membranes. AB - The coronavirus replicase gene (gene 1) is translated into two co-amino-terminal polyproteins that are proteolytically processed to yield more than 15 mature proteins. Several gene 1 proteins have been shown to localize at sites of viral RNA synthesis in the infected cell cytoplasm, notably on late endosomes at early times of infection. However, both immunofluorescence and electron microscopic studies have also detected gene 1 proteins at sites distinct from the putative sites of viral RNA synthesis or virus assembly. In this study, mouse hepatitis virus (MHV)-infected cells were fractionated and analyzed to determine if gene 1 proteins segregated to more than one membrane population. Following differential centrifugation of lysates of MHV-infected DBT cells, gene 1 proteins as well as the structural N and M proteins were detected almost exclusively in a high-speed small membrane pellet. Following fractionation of the small membrane pellet on an iodixanol density gradient, the gene 1 proteins p28 and helicase cofractionated with dense membranes (1.12 to 1.13 g/ml) that also contained peak concentrations of N. In contrast, p65 and p1a-22 were detected in a distinct population of less dense membranes (1.05 to 1.09 g/ml). Viral RNA was detected in membrane fractions containing helicase, p28, and N but not in the fractions containing p65 and p1a 22. LAMP-1, a marker for late endosomes and lysosomes, was detected in both membrane populations. These results demonstrate that multiple gene 1 proteins segregate into two biochemically distinct but tightly associated membrane populations and that only one of these populations appears to be a site for viral RNA synthesis. The results further suggest that p28 is a component of the viral replication complex whereas the gene 1 proteins p1a-22 and p65 may serve roles during infection that are distinct from viral RNA transcription or replication. PMID- 10823873 TI - Isolation and characterization of a new subtype of Borna disease virus. AB - Borna disease virus (BDV), the causative agent of severe meningoencephalitis in a wide variety of animal species, has been considered to be genetically invariable and to form a single type within the genus Bornavirus of the family Bornaviridae. BDV infections are of particular interest, because for the first time a virus infection appears to be linked to human psychiatric disorders. We now describe a new subtype of BDV isolated from a horse which was euthanatized due to severe, incurable neurological disease. The nucleotide sequence of this new strain, named No/98, differs from the reference strains by more than 15%, and the subtype is difficult to detect by standard reverse transcriptase PCR protocols. The nucleotide exchanges of the novel BDV isolate have surprisingly little effect on the primary structures of most viral proteins, with the notable exception of the X protein (p10), which is only 81% identical to its counterpart in reference strains. Our data indicate that the genome of BDV is far more variable than previously assumed and that naturally occurring subtypes may escape detection by currently used diagnostic assays. PMID- 10823874 TI - Conserved surface-exposed K/R-X-K/R motifs and net positive charge on poxvirus complement control proteins serve as putative heparin binding sites and contribute to inhibition of molecular interactions with human endothelial cells: a novel mechanism for evasion of host defense. AB - Vaccinia virus complement control protein (VCP) has been shown to possess the ability to inhibit both classical and alternative complement pathway activation. The newly found ability of this protein to bind to heparin has been shown in previous studies to result in uptake by mast cells, possibly promoting tissue persistence. It has also been shown to reduce chemotactic migration of leukocytes by blocking chemokine binding. In addition, this study shows that VCP-through its ability to bind to glycosaminoglycans (heparin-like molecules) on the surface of human endothelial cells-is able to block antibody binding to surface major histocompatibility complex class I molecules. Since heparin binding is critical for many functions of this protein, we have attempted to characterize the molecular basis for this interaction. Segments of this protein, generated by genetic engineering of the DNA encoding VCP into the Pichia pastoris expression system, were used to localize the regions with heparin binding activity. These regions were then analyzed to more specifically define their properties for binding. It was found that the number of putative binding sites (K/R-X-K/R), the overall positive charge, and the percentage of positively charged amino acids within the protein were responsible for this interaction. PMID- 10823875 TI - The surface conformation of Sindbis virus glycoproteins E1 and E2 at neutral and low pH, as determined by mass spectrometry-based mapping. AB - Sindbis virus contains two membrane glycoproteins, E1 and E2, which are organized into 80 trimers of heterodimers (spikes). These trimers form a precise T=4 icosahedral protein lattice on the surface of the virus. Very little is known about the organization of the E1 and E2 glycoproteins within the spike trimer. To gain a better understanding of how the proteins E1 and E2 are arranged in the virus membrane, we have used the techniques of limited proteolysis and amino acid chemical modification in combination with mass spectrometry. We have determined that at neutral pH the E1 protein regions that are accessible to proteases include domains 1-21 (region encompassing amino acids 1 to 21), 161-176, and 212 220, while the E2 regions that are accessible include domains 31-84, 134-148, 158 186, 231-260, 299-314, and 324-337. When Sindbis virus is exposed to low pH, E2 amino acid domains 99-102 and 262-309 became exposed while other domains became inaccessible. Many new E1 regions became accessible after exposure to low pH, including region 86-91, which is in the putative fusion domain of E1 of Semliki Forest virus (SFV) (M. C. Kielian et al., J. Cell Biol. 134:863-872, 1996). E1 273-287 and region 145-158 were also exposed at low pH. These data support a model for the structure of the alphavirus spike in which the E1 glycoproteins are centrally located as trimers which are surrounded and protected by the E2 glycoprotein. These data improve our understanding of the structure of the virus membrane and have implications for understanding the protein conformational changes which accompany the process of virus-cell membrane fusion. PMID- 10823876 TI - Differential narrow focusing of immunodominant human immunodeficiency virus gag specific cytotoxic T-lymphocyte responses in infected African and caucasoid adults and children. AB - Cytotoxic T-lymphocyte (CTL) activity plays a central role in control of viral replication and in determining outcome in cases of human immunodeficiency virus type 1 (HIV-1) infection. Incorporation of important CTL epitope sequences into candidate vaccines is, therefore, vital. Most CTL studies have focused upon small numbers of adult Caucasoid subjects infected with clade-B virus, whereas the global epidemic is most severe in sub-Saharan African populations and predominantly involves clade-C infection in both adults and children. In this study, sensitive enzyme-linked immunospot (elispot) assays have been utilized to identify the dominant Gag-specific CTL epitopes targeted by adults and children infected with clade-B or -C virus. Cohorts evaluated included 44 B-clade-infected Caucasoid American and African American adults and children and 37 C-clade infected African adults and children from Durban, South Africa. The results show that 3 out of 46 peptides spanning p17(Gag) and p24(Gag) sequences tested contain two-thirds of the dominant Gag-specific epitopes, irrespective of the clade, ethnicity, or age group studied. However, there were distinctive differences between the dominant responses made by Caucasoids and Africans. Dominant responses in Caucasoids were more often within p17(Gag) peptide residues 16 to 30 (38 versus 12%; P < 0.01), while p24(Gag) peptide residues 41 to 60 contained the dominant Gag epitope more often in the African subjects tested (39 versus 4%; P < 0.005). Within this 20-mer p24(Gag), an epitope presented by both B42 and B81 is defined which represents the dominant Gag response in >30% of the total infected population in Durban. This epitope is closely homologous with dominant HIV-2 and simian immunodeficiency virus Gag-specific CTL epitopes. The fine focusing of dominant CTL responses to these few regions of high immunogenicity is of significance to vaccine design. PMID- 10823879 TI - Application of the intracellular gamma interferon assay to recalculate the potency of CD8(+) T-cell responses to herpes simplex virus. AB - Enumeration and characterization of herpes simplex virus (HSV)-specific CD8(+) T lymphocytes are tedious and indirect. We quantitated antigen-specific CD8(+) T cells during acute and secondary stages of HSV infection using intracellular gamma interferon production upon stimulation with virus or immunodominant peptide. Results show a substantial increase in the number of CD8(+) T cells which was otherwise underestimated with the conventional limiting dilution analysis. PMID- 10823878 TI - Simian immunodeficiency viruses of diverse origin can use CXCR4 as a coreceptor for entry into human cells. AB - Primary simian immunodeficiency virus (SIV) isolated from sooty mangabey (SIVsm [n = 6]), stumptail (SIVstm [n = 1]), mandrill (SIVmnd [n = 1]), and African green (SIVagm [n = 1]) primates were examined for their ability to infect human cells and for their coreceptor requirements. All isolates infected human peripheral blood mononuclear cells (PBMCs) from a CCR5(+/+) donor, and seven of eight isolates tested also infected CCR5(-/-) PBMCs. Analysis of coreceptor utilization using GHOST and U87 cell lines revealed that all of the isolates tested used CCR5 and the orphan receptors STRL33 and GPR15. Coreceptors such as CCR2b, CCR3, CCR8, and CX3CR1 were also utilized by some primary SIV isolates. More importantly, we found that CXCR4 was used as a coreceptor by the SIVstm, the SIVagm, and four of the SIVsm isolates in GHOST and U87 cells. These data suggest that primary SIV isolates from diverse primate species can utilize CXCR4 for viral entry, similar to what has been described for human immunodeficiency viruses. PMID- 10823877 TI - Simian and human immunodeficiency virus Nef proteins use different surfaces to downregulate class I major histocompatibility complex antigen expression. AB - Simian immunodeficiency virus (SIV) and human immunodeficiency virus type 1 (HIV 1) Nef proteins are related regulatory proteins that share several functions, including the ability to downregulate class I major histocompatibility complex (MHC) and CD4 expression on the cell surface and to alter T-cell-receptor initiated signal transduction in T cells. We compared the mechanisms used by SIV mac239 Nef and HIV-1 Nef to downregulate class I MHC and found that the ability of SIV Nef to downregulate class I MHC requires a unique C-terminal region of the SIV mac239 Nef molecule which is not found in HIV-1 Nef. Interestingly, mutation of the PxxP motif in SIV Nef, unlike in HIV-1 Nef, does not affect class I MHC downregulation. We also found that downregulation of class I MHC by SIV Nef requires a conserved tyrosine in the cytoplasmic domain of the class I MHC heavy chain and involves accelerated endocytosis of class I complexes, as previously found with HIV-1 Nef. Thus, while SIV and HIV-1 Nef proteins use a similar mechanism to downregulate class I MHC expression, they have evolved different surfaces for molecular interactions with cell factors that regulate class I MHC traffic. Mutations in the C-terminal domain of SIV mac239 Nef selectively disrupt class I MHC downregulation, having no detectable effect on other functions of Nef, such as the downregulation of CD4 and CD3 surface expression, the stimulation of SIV virion infectivity, and the induction of SIV replication from T cells infected in the absence of stimulation. The resulting mutants will be useful reagents for studying the importance of class I MHC downregulation for SIV replication and AIDS pathogenesis in infected rhesus macaques. PMID- 10823880 TI - Design and intracellular activity of a human single-chain antibody to human immunodeficiency virus type 1 conserved gp41 epitope. AB - A human lymphoid cell line (F172-D8) excreting a human immunodeficiency virus type 1 (HIV-1) anti-gp41 monoclonal antibody was used to construct a plasmid containing the cDNA of the single-chain variable fragment (scFvD8) corresponding to this antibody. A stable human osteosarcoma cell line was obtained which expressed the scFvD8 protein in the cytoplasm. Whereas a cell line transfected with a control construct (pCI-neo) was readily and productively infected with laboratory (Ba-L) or primary HIV-1 isolates, the scFvD8 cell line did not support productive infection. Binding of the virus, internalization, and reverse transcription were not altered by scFvD8 expression, but gp160 expression was dramatically reduced. These data suggest that cytoplasmic expression of this artificial single-chain antibody can interfere with gp160 expression, thereby reducing the production of mature viral envelope proteins. PMID- 10823881 TI - Characterization of stable, soluble trimers containing complete ectodomains of human immunodeficiency virus type 1 envelope glycoproteins. AB - The human immunodeficiency virus type 1 (HIV-1) envelope glycoproteins function as a membrane-anchored trimer of three gp120 exterior glycoproteins and three gp41 transmembrane glycoproteins. Previously, we reported three approaches to stabilize soluble trimers containing parts of the gp41 ectodomains: addition of GCN4 trimeric helices, disruption of the cleavage site between gp120 and gp41, and introduction of cysteines in the gp41 coiled coil to form intersubunit disulfide bonds. Here, we applied similar approaches to stabilize soluble gp140 trimers including the complete gp120 and gp41 ectodomains. A combination of fusion with the GCN4 trimeric sequences and disruption of the gp120-gp41 cleavage site resulted in relatively homogeneous gp140 trimers with exceptional stability. The gp120 epitopes recognized by neutralizing antibodies are intact and exposed on these gp140 trimers. By contrast, the nonneutralizing antibody epitopes on the gp120 subunits of the soluble trimers are relatively occluded compared with those on monomeric gp120 preparations. This antigenic similarity to the functional HIV 1 envelope glycoproteins and the presence of the complete gp41 ectodomain should make the soluble gp140 trimers useful tools for structural and immunologic studies. PMID- 10823882 TI - Unwinding of a herpes simplex virus type 1 origin of replication (Ori(S)) by a complex of the viral origin binding protein and the single-stranded DNA binding protein. AB - A herpes simplex virus type 1 (HSV-1) Ori(S) analogue in which the A+T sequence linking the box I and II elements was replaced by two single-stranded oligo(dT)s is unwound by the UL9 protein-ICP8 complex. Unwinding of wild-type Ori(S) by the UL9 protein-ICP8 complex was also observed under conditions which destabilize the A+T sequence. These experiments support a model for the unwinding of Ori(S) in which destabilization of the A+T sequence can generate a single-stranded DNA binding site for ICP8, which then associates with the UL9 protein bound to boxes I and II to promote the bidirectional unwinding of Ori(S). PMID- 10823883 TI - Impact of human immunodeficiency virus type 1 RNA dimerization on viral infectivity and of stem-loop B on RNA dimerization and reverse transcription and dissociation of dimerization from packaging. AB - The kissing-loop domain (KLD) encompasses a stem-loop, named kissing-loop or dimerization initiation site (DIS) hairpin (nucleotides [nt] 248 to 270 in the human immunodeficiency virus type 1 strains HIV-1(Lai) and HIV-1(Hxb2)), seated on top of a 12-nt stem-internal loop called stem-loop B (nt 243 to 247 and 271 to 277). Destroying stem-loop B reduced genome dimerization by approximately 50% and proviral DNA synthesis by approximately 85% and left unchanged the dissociation temperature of dimeric genomic RNA. The most affected step of reverse transcription was plus-strand DNA transfer, which was reduced by approximately 80%. Deleting nt 241 to 256 or 200 to 256 did not reduce genome dimerization significantly more than the destruction of stem-loop B or the DIS hairpin. We conclude that the KLD is nonmodular: mutations in stem-loop B and in the DIS hairpin have similar effects on genome dimerization, reverse transcription, and encapsidation and are also "nonadditive"; i.e., a larger deletion spanning both of these structures has the same effects on genome dimerization and encapsidation as if stem-loop B strongly impacted DIS hairpin function and vice versa. A C258G transversion in the palindrome of the kissing-loop reduced genome dimerization by approximately 50% and viral infectivity by approximately 1.4 log. Two mutations, CGCG261-->UUAA261 (creating a weaker palindrome) and a Delta241-256 suppressor mutation, were each able to reduce genome dimerization but leave genome packaging unaffected. PMID- 10823884 TI - Maternal SDF1 3'A polymorphism is associated with increased perinatal human immunodeficiency virus type 1 transmission. AB - Genetic polymorphisms in chemokine and chemokine receptor genes influence susceptibility to human immunodeficiency virus type 1 (HIV-1) infection and disease progression, but little is known regarding the association between these allelic variations and the ability of the host to transmit virus. In this study, we show that the maternal heterozygous SDF1 genotype (SDF1 3'A/wt) is associated with perinatal transmission of HIV-1 (risk ratio [RR], 1.8; 95% confidence interval [CI], 1.0 to 3.3) and particularly postnatal breastmilk transmission (RR, 3.1; 95% CI, 1.1 to 8.6). In contrast, the infant SDF1 genotype had no effect on mother-to-infant transmission. These data suggest that SDF1, which is a ligand for the T-tropic HIV-1 coreceptor CXCR4, may affect the ability of a mother to transmit the virus to her infant. This suggests that a genetic polymorphism in a gene encoding a chemokine receptor ligand may be associated with increased infectivity of the index case and highlights the importance of considering transmission as well as clinical outcome in designing chemokine-based therapies for HIV-1. PMID- 10823885 TI - Prostaglandin E(2) increases bovine leukemia virus tax and pol mRNA levels via cyclooxygenase 2: regulation by interleukin-2, interleukin-10, and bovine leukemia virus. AB - Prostaglandin E(2) (PGE(2)), produced by macrophages, has important immune regulatory functions, suppressing a type 1 immune response and stimulating a type 2 immune response. Type 1 cytokines (interleukin-2 [IL-2], IL-12, and gamma interferon) increase in freshly isolated peripheral blood mononuclear cells (PBMCs) of animals with an early disease stage of bovine leukemia virus (BLV) infection, while IL-10 increases in animals with a late disease stage. Although IL-10 has an immunosuppressive role in the host immune system, IL-10 also inhibits BLV tax and pol mRNA levels in vitro. In contrast, IL-2 stimulates BLV tax and pol mRNA and p24 protein expression in cultured PBMCs. The inhibitory effect of IL-10 on BLV expression depends on soluble factors secreted by macrophages. Thus, we hypothesized that PGE(2), a cyclooxygenase 2 (COX-2) product of macrophages, may regulate BLV expression. Here, we show that the level of COX-2 mRNA was decreased in PBMCs treated with IL-10, while IL-2 enhanced the level of COX-2 mRNA. Addition of PGE(2) stimulated BLV tax and pol mRNA levels and reversed the IL-10 inhibition of BLV mRNA. In addition, the specific COX-2 inhibitor, NS-398, inhibited the amount of BLV mRNA detected. Addition of PGE(2) increased BLV tax mRNA regardless of NS-398 addition. PGE(2) inhibited antigen specific PBMC stimulation, suggesting that stimulation of BLV tax and pol mRNA levels by PGE(2) is independent of cell proliferation. These findings suggest that macrophage-derived COX-2 products, such as PGE(2), regulate virus expression and disease progression in BLV infection. PMID- 10823886 TI - Transforming growth factor beta -inducible independent binding of SMAD to the Smad7 promoter. AB - SMAD proteins can mediate transforming growth factor beta (TGF-beta)-inducible transcriptional responses. Whereas SMAD can recognize specific DNA sequences, it is usually recruited to a promoter through interaction with a DNA-binding partner. In an effort to search for TGF-beta-inducible genes, we used a subtractive screening method and identified human Smad7, which can antagonize TGF beta signaling and is rapidly up-regulated by TGF-beta. In this report, we show that TGF-beta can stabilize Smad7 mRNA and activate Smad7 transcription. The Smad7 promoter is the first TGF-beta responsive promoter identified in vertebrates that contains the 8-bp palindromic SMAD-binding element (SBE), an optimal binding site previously identified by a PCR-based selection from random oligonucleotides by using recombinant Smad3 and Smad4. We demonstrate that on TGF beta treatment, endogenous SMAD complex can bind to a Smad7 promoter DNA as short as 14 or 16 bp that contains the 8-bp SBE in gel mobility shift and supershift assays. Our studies provide strong evidence that SMAD proteins can bind to a natural TGF-beta responsive promoter independent of other sequencespecific transcription factors. We further show that, whereas recombinant Smad3 binds to the SBE, endogenous or even transfected Smad3 cannot bind to the SBE in the absence of Smad4. These findings have important implications in the identification of target genes of the TGF-beta/SMAD signaling pathways. PMID- 10823887 TI - Conserved and divergent aspects of terminal patterning in the beetle Tribolium castaneum. AB - To infer similarities and differences in terminal pattern formation in insects, we analyzed several of the key genes of this process in the beetle Tribolium castaneum. We cloned two genes of the terminal pattern cascade, namely tailless (tll) and forkhead (fkh), from Tribolium and studied their expression patterns. In addition, we analyzed the pattern of MAP kinase activation at blastoderm stage as a possible signature for torso-dependent signaling. Further, we analyzed the late expression of the previously cloned Tribolium caudal (Tc-cad) gene. Finally, we used the upstream region of Tc-tll to drive a reporter gene construct in Drosophila. We find that this construct is activated at the terminal regions in Drosophila, suggesting that the torso-dependent pathway is conserved between the species. We show that most of the expression patterns of the genes studied here are similar in Drosophila and Tribolium, suggesting conserved functions. There is, however, one exception, namely the early function of Tc-tll at the posterior pole. In Drosophila, the posterior tll expression is involved in the direct regulation of the target genes of the terminal pathway. In Tribolium, posterior Tc-tll expression occurs only for a short time and ceases before the target genes known from Drosophila are activated. Thus, we infer that Tc-tll does not function as a direct regulator of segmentation genes at the posterior end. It is more likely to be involved in the early specification of a group of "terminal" cells, which begin to differentiate only at a later stage of embryogenesis, when much of the abdominal segmentation process is complete. Thus, there appears to have been a major shift in tll function during the evolutionary transition from short germ to long germ embryogenesis. PMID- 10823888 TI - Regulation of the actin cycle in vivo by actin filament severing. AB - Cycling of actin subunits between monomeric and filamentous phases is essential for cell crawling behavior. We investigated actin filament turnover rates, length, number, barbed end exposure, and binding of cofilin in bovine arterial endothelial cells moving at different speeds depending on their position in a confluent monolayer. Fast-translocating cells near the wound edge have short filament lifetimes compared with turnover values that proportionately increase in slower moving cells situated at increasing distances from the wound border. Contrasted with slow cells exhibiting slow actin filament turnover speeds, fast cells have less polymerized actin, shorter actin filaments, more free barbed ends, and less actin-associated cofilin. Cultured primary fibroblasts manifest identical relationships between speed and actin turnover as the endothelial cells, and fast fibroblasts expressing gelsolin have higher actin turnover rates than slow fibroblasts that lack this actin-severing protein. These results implicate actin filament severing as an important control mechanism for actin cycling in cells. PMID- 10823889 TI - Simultaneous activation of NADPH oxidase-related proton and electron currents in human neutrophils. AB - Generation of reactive oxygen species by the NADPH oxidase complex is an important bactericidal weapon of phagocytes. Phorbol myristate acetate (PMA) is a potent agonist for this "respiratory burst" in human neutrophils. Although stoichiometric H(+) efflux occurs during the respiratory burst, efforts to stimulate voltage-gated H(+) channels by PMA in whole-cell patch-clamped phagocytes have been unsuccessful. We have used a modification of the permeabilized-patch configuration that allows control of intracellular pH and preserves second-messenger pathways. Using this method, we show that PMA dramatically enhances and alters voltage-gated proton currents in human neutrophils. PMA produced four alterations in H(+) current properties, each of which increases the H(+) current at any given voltage: (i) a 40-mV negative shift in the H(+) conductance-voltage (g(H)-V) relationship; (ii) faster activation [smaller activation time constant (tau(act))] during depolarizing pulses; (iii) slower deactivation [larger deactivation time constant (tau(tail))] on repolarization; and (iv) a larger maximum H(+) conductance (g(H, max)). Inward current that directly reflects electron transport by NADPH oxidase was also activated by PMA stimulation. The identity of this electron current was confirmed by its sensitivity to diphenylene iodinium, an inhibitor of NADPH oxidase. Diphenylene iodinium also reversed the slowing of tau(tail) with a time course paralleling the inhibition of electron current. However, the amplitudes of H(+) and electron currents activated by PMA were not correlated. A complex interaction between NADPH oxidase and voltage-gated proton channels is indicated. The data suggest that PMA stimulation modulates preexisting H(+) channels rather than inducing a new H(+) channel. PMID- 10823890 TI - Selection-mutation balance in polysomic tetraploids: impact of double reduction and gametophytic selection on the frequency and subchromosomal localization of deleterious mutations. AB - We modeled the behavior of recessive mutations with deleterious effects to either the sporophyte or the gametophyte, or both, in polysomic tetraploid populations by allowing for varying levels of double reduction, mutation, and self fertilization. Double reduction causes a decrease of the equilibrium frequencies of deleterious alleles, and it has much more influence on genes subjected to gametophytic selection than on genes solely under sporophytic selection. With gametophytic selection, low frequencies of double reduction are enough to reduce equilibrium frequencies severalfold. Double reduction occurs when sister alleles migrate to the same gamete during meiosis. It depends on the frequency at which a locus recombines with its centromere, and on the frequency of multivalent formation. Therefore, a greater accumulation of deleterious mutations should occur on polysomic chromosomes with a prevalence of bivalent pairing and in chromosomal regions between centromeres and proximal chiasmata. Proximal loci should have a greater impact in reducing the fitness of a polyploid population being inbred. This prediction can explain observations that homozygosities at different subchromosomal regions have distinct effects on inbreeding depression in polyploids. Furthermore, even mildly deleterious alleles can lead to large amounts of inbreeding depression because of their high equilibrium frequencies. Molecular studies correlating level of heterozygosity and degree of heterosis should take into account this nonuniform distribution of deleterious alleles in polyploid genomes. Preservation or enhancement of heterozygosity would be more critical at proximal regions than at other chromosome regions in polysomic polyploid species. PMID- 10823891 TI - The Huntington's disease protein interacts with p53 and CREB-binding protein and represses transcription. AB - Huntington's Disease (HD) is caused by an expansion of a polyglutamine tract within the huntingtin (htt) protein. Pathogenesis in HD appears to include the cytoplasmic cleavage of htt and release of an amino-terminal fragment capable of nuclear localization. We have investigated potential consequences to nuclear function of a pathogenic amino-terminal region of htt (httex1p) including aggregation, protein-protein interactions, and transcription. httex1p was found to coaggregate with p53 in inclusions generated in cell culture and to interact with p53 in vitro and in cell culture. Expanded httex1p represses transcription of the p53-regulated promoters, p21(WAF1/CIP1) and MDR-1. httex1p was also found to interact in vitro with CREB-binding protein (CBP) and mSin3a, and CBP to localize to neuronal intranuclear inclusions in a transgenic mouse model of HD. These results raise the possibility that expanded repeat htt causes aberrant transcriptional regulation through its interaction with cellular transcription factors which may result in neuronal dysfunction and cell death in HD. PMID- 10823892 TI - Unfolding proteins by external forces and temperature: the importance of topology and energetics. AB - Unfolding of proteins by forced stretching with atomic force microscopy or laser tweezer experiments complements more classical techniques using chemical denaturants or temperature. Forced unfolding is of particular interest for proteins that are under mechanical stress in their biological function. For beta sandwich proteins (a fibronectin type III and an immunoglobulin domain), both of which appear in the muscle protein titin, the results of stretching simulations show important differences from temperature-induced unfolding, but there are common features that point to the existence of folding cores. Intermediates detected by comparing unfolding with a biasing perturbation and a constant pulling force are not evident in temperature-induced unfolding. For an alpha helical domain (alpha-spectrin), which forms part of the cytoskeleton, there is little commonality in the pathways from unfolding induced by stretching and temperature. Comparison of the forced unfolding of the two beta-sandwich proteins and two alpha-helical proteins (the alpha-spectrin domain and an acyl-coenzyme A binding protein) highlights important differences within and between protein classes that are related to the folding topologies and the relative stability of the various structural elements. PMID- 10823893 TI - Phosphoinositide-3 kinase binds to a proline-rich motif in the Na+, K+-ATPase alpha subunit and regulates its trafficking. AB - Endocytosis of Na(+),K(+)-ATPase molecules in response to G protein-coupled receptor stimulation requires activation of class I(A) phosphoinositide-3 kinase (PI3K-I(A)) in a protein kinase C-dependent manner. In this paper, we report that PI3K-I(A), through its p85alpha subunit-SH3 domain, binds to a proline-rich region in the Na(+),K(+)-ATPase catalytic alpha subunit. This interaction is enhanced by protein kinase C-dependent phosphorylation of a serine residue that flanks the proline-rich motif in the Na(+),K(+)-ATPase alpha subunit and results in increased PI3K-I(A) activity, an effect necessary for adaptor protein 2 binding and clathrin recruitment. Thus, Ser-phosphorylation of the Na(+),K(+) ATPase catalytic subunit serves as an anchor signal for regulating the location of PI3K-I(A) and its activation during Na(+),K(+)-ATPase endocytosis in response to G protein-coupled receptor signals. PMID- 10823894 TI - Dynamic actin filaments are required for stable long-term potentiation (LTP) in area CA1 of the hippocampus. AB - The hypothesis that dynamic actin filaments participate in specific aspects of synaptic plasticity was investigated at the Schaffer-collateral-CA1 pyramidal cell synapse of mouse hippocampus. Low concentrations (0.01-1 microM) of compounds that inhibit actin filament assembly were bath applied to hippocampal slices during extracellular recording of field excitatory postsynaptic potentials. Cytochalasin D, cytochalasin B, and latrunculin A all impaired the maintenance of LTP induced by brief high-frequency stimulation. This effect on LTP maintenance was specific, because none of the compounds affected basal synaptic transmission, paired-pulse facilitation, LTP induction, or post-tetanic potentiation. The effect of cytochalasin B was reversible. The results are consistent with a model in which dynamic actin filaments play an essential role in the molecular mechanisms underlying the early maintenance phase of LTP, such as growth of new synaptic connections or conversion of silent synapses. PMID- 10823895 TI - Characterization of the 1918 "Spanish" influenza virus neuraminidase gene. AB - The "Spanish" influenza pandemic of 1918 was characterized by exceptionally high mortality, especially among young adults. The surface proteins of influenza viruses, hemagglutinin and neuraminidase, play important roles in virulence, host specificity, and the human immune response. The complete coding sequence of hemagglutinin was reported last year. This laboratory has now determined the complete coding sequence of the neuraminidase gene of the 1918 virus. Influenza RNA fragments were isolated from lung tissue of three victims of the 1918 flu; complete sequence was generated from A/Brevig Mission/1/18, with confirmatory sequencing carried out on A/South Carolina/1/18 and A/New York/1/18. The 1918 neuraminidase gene sequence was compared with other N1 subtype neuraminidase genes, including 9 N1 strains newly sequenced for this study. The 1918 neuraminidase shares many sequence and structural characteristics with avian strains, including the conserved active site, wild-type stalk length, glycosylation sites, and antigenic sites. Phylogenetically, the 1918 neuraminidase gene appears to be intermediate between mammals and birds, suggesting that it was introduced into mammals just before the 1918 pandemic. PMID- 10823896 TI - CpG methylation as a mechanism for the regulation of E2F activity. AB - Regulation of gene expression in mammals through methylation of cytosine residues at CpG dinucleotides is involved in the development and progression of tumors. Because many genes that are involved in the control of cell proliferation are regulated by members of the E2F family of transcription factors and because some E2F DNA-binding sites are methylated in vivo, we have investigated whether CpG methylation can regulate E2F functions. We show here that methylation of E2F elements derived from the dihydrofolate reductase, E2F1, and cdc2 promoters prevents the binding of all E2F family members tested (E2F1 through E2F5). In contrast, methylation of the E2F elements derived from the c-myc and c-myb promoters minimally affects the binding of E2F2, E2F3, E2F4, and E2F5 but significantly inhibits the binding of E2F1. Consistent with these studies, E2F3, but not E2F1, activates transcription through methylated E2F sites derived from the c-myb and c-myc genes whereas both E2F1 and E2F3 fail to transactivate a reporter gene that is under the control of a methylated dihydrofolate reductase E2F site. Together, these data illustrate a means through which E2F activity can be controlled. PMID- 10823897 TI - The Bloom's syndrome gene product promotes branch migration of holliday junctions. AB - Bloom's syndrome (BS) is an autosomal recessive disorder associated with dwarfism, immunodeficiency, reduced fertility, and elevated levels of many types of cancer. BS cells show marked genomic instability; in particular, hyperrecombination between sister chromatids and homologous chromosomes. This instability is thought to result from defective processing of DNA replication intermediates. The gene mutated in BS, BLM, encodes a member of the RecQ family of DExH box DNA helicases, which also includes the Werner's syndrome gene product. We have investigated the mechanism by which BLM suppresses hyperrecombination. Here, we show that BLM selectively binds Holliday junctions in vitro and acts on recombination intermediates containing a Holliday junction to promote ATP-dependent branch migration. We present a model in which BLM disrupts potentially recombinogenic molecules that arise at sites of stalled replication forks. Our results have implications for the role of BLM as an anti recombinase in the suppression of tumorigenesis. PMID- 10823898 TI - Control of protein crystal nucleation around the metastable liquid-liquid phase boundary. AB - The capability to enhance or suppress the nucleation of protein crystals opens opportunities in various fundamental and applied areas, including protein crystallography, production of protein crystalline pharmaceuticals, protein separation, and treatment of protein condensation diseases. Herein, we show that the rate of homogeneous nucleation of lysozyme crystals passes through a maximum in the vicinity of the liquid-liquid phase boundary hidden below the liquidus (solubility) line in the phase diagram of the protein solution. We found that glycerol and polyethylene glycol (which do not specifically bind to proteins) shift this phase boundary and significantly suppress or enhance the crystal nucleation rates, although no simple correlation exists between the action of polyethylene glycol on the phase diagram and the nucleation kinetics. The control mechanism does not require changes in the protein concentration, acidity, and ionicity of the solution. The effects of the two additives on the phase diagram strongly depend on their concentration, which provides opportunities for further tuning of nucleation rates. PMID- 10823899 TI - Amphetamine-induced loss of human dopamine transporter activity: an internalization-dependent and cocaine-sensitive mechanism. AB - The dopamine transporter (DAT) is a target of amphetamine (AMPH) and cocaine. These psychostimulants attenuate DAT clearance efficiency, thereby increasing synaptic dopamine (DA) levels. Re-uptake rate is determined by the number of functional transporters at the cell surface as well as by their turnover rate. Here, we present evidence that DAT substrates, including AMPH and DA, cause internalization of human DAT, thereby reducing transport capacity. Acute treatment with AMPH reduced the maximal rate of [(3)H]DA uptake, decreased AMPH induced currents, and significantly redistributed the immunofluorescence of an epitope-tagged DAT from the plasma membrane to the cytosol in human embryonic kidney 293 cells. Conversely, DAT inhibitors, such as cocaine, mazindol, and nomifensine, when administered with AMPH, blocked the reduction in [(3)H]DA uptake and the redistribution of DAT immunofluorescence to the cytosol. The reductions of [(3)H]DA uptake and AMPH-induced DAT internalization also were inhibited by coexpression of a dominant negative mutant of dynamin I (K44A), indicating that endocytosis modulates transport capacity, likely through a clathrin-mediated pathway. With this mechanism of regulation, acute application of AMPH would reduce DA uptake not only by direct competition for uptake, but also by reducing the available cell-surface DAT. Moreover, AMPH-induced internalization might diminish the amount of DAT available for DA efflux, thereby modulating the cytotoxic effects of elevated extracellular DA. PMID- 10823900 TI - A potent dimeric peptide antagonist of interleukin-5 that binds two interleukin-5 receptor alpha chains. AB - Two series of peptides that specifically bind to the extracellular domain of the alpha chain of the human interleukin-5 receptor (IL-5Ralpha), but share no primary sequence homology to IL-5, were identified from libraries of random recombinant peptides. Affinity maturation procedures generated a 19-aa peptide that binds to the IL-5 receptor alpha/beta heterodimer complex with an affinity equal to that of IL-5 and is a potent and specific antagonist of IL-5 activity in a human eosinophil adhesion assay. The active form of the peptide is a disulfide crosslinked dimer that forms spontaneously in solution. Gel filtration analysis, receptor-binding studies, and analytical ultracentrifugation reveal that the dimeric peptide binds simultaneously to two receptor alpha chains in solution. Furthermore, the dimer peptide, but not IL-5, can activate a chimeric receptor consisting of the IL-5Ralpha extracellular domain fused to the intracellular domain of the epidermal growth factor receptor, thus demonstrating that the peptide also promotes receptor dimerization in a cellular context. The functional antagonism produced by the bivalent interaction of the dimeric peptide with two IL-5R alpha chains represents a distinctive mechanism for the antagonism of cytokines that use heteromeric receptors. PMID- 10823902 TI - Subjective surfaces: a method for completing missing boundaries. AB - We present a model and algorithm for segmentation of images with missing boundaries. In many situations, the human visual system fills in missing gaps in edges and boundaries, building and completing information that is not present. This presents a considerable challenge in computer vision, since most algorithms attempt to exploit existing data. Completion models, which postulate how to construct missing data, are popular but are often trained and specific to particular images. In this paper, we take the following perspective: We consider a reference point within an image as given and then develop an algorithm that tries to build missing information on the basis of the given point of view and the available information as boundary data to the algorithm. We test the algorithm on some standard images, including the classical triangle of Kanizsa and low signal/noise ratio medical images. PMID- 10823901 TI - Combined effects of radiotherapy and angiostatin gene therapy in glioma tumor model. AB - The objective of the present study was to evaluate the antitumor effect of a defective adenovirus expressing a secretable angiostatin-like molecule (AdK3) in combination with radiotherapy in rat C6 gliomas s.c. preestablished into athymic mice. In vitro, the combination regimen was significantly (P < 0.001) more cytotoxic for human microcapillary endothelial cells than either treatment alone, whereas survival of C6 glioma cells was not affected in the conditions used. Radiotherapy and AdK3 gene delivery was then studied on well established C6 xenografts (165 +/- 70 mm(3)). In these tumors, AdK3 intratumoral injections had only a marginal effect. Interestingly, when experimental radiotherapy was added, significantly higher (P < 0.005), and possibly synergistic, antitumoral effects were observed that tightly correlated a marked decrease of intratumoral vascularization. The combination of radiotherapy and AdK3 intratumoral injections also revealed a significant (P < 0.05) inhibition of tumor growth as compared with either treatment alone for larger tumors (467 +/- 120 mm(3)). Altogether, these data emphasize the potential of combining a destructive strategy directed against the tumor cells with an anti-angiogenic approach to fight cancer. PMID- 10823903 TI - A mechanistic basis for Mre11-directed DNA joining at microhomologies. AB - Repair of DNA double-strand breaks in vertebrate cells occurs mainly by an end joining process that often generates junctions with sequence homologies of a few nucleotides. Mre11 is critical for this mode of repair in budding yeast and has been implicated in the microhomology-based joining. Here, we show that Mre11 exonuclease activity is sensitive to the presence of heterologous DNA, and to the structure and sequence of its ends. Addition of mismatched DNA ends stimulates degradation of DNA by Mre11, whereas cohesive ends strongly inhibit it. Furthermore, if a sequence identity is revealed during the course of degradation, it causes Mre11 nuclease activity to pause, thus stabilizing the junction at a site of microhomology. A nuclease-deficient Mre11 mutant that still binds DNA can also stimulate degradation by wild-type Mre11, suggesting that Mre11-DNA complexes may interact to bridge DNA ends and facilitate DNA joining. PMID- 10823904 TI - Decoupled evolution of coding region and mRNA expression patterns after gene duplication: implications for the neutralist-selectionist debate. AB - The neutralist perspective on molecular evolution maintains that the vast majority of mutations affecting gene function are neutral or deleterious. After a gene duplication where both genes are retained, it predicts that original and duplicate genes diverge at clock-like rates. This prediction is usually tested for coding sequences, but can also be applied to another important aspect of gene function, the genes' expression pattern. Moreover, if both sequence and expression pattern diverge at clock-like rates, a correlation between divergence in sequence and divergence in expression patterns is expected. Duplicate gene pairs with more highly diverged sequences should also show more highly diverged expression patterns. This prediction is tested for a large sample of duplicated genes in the yeast Saccharomyces cerevisiae, using both genome sequence and microarray expression data. Only a weak correlation is observed, suggesting that coding sequence and mRNA expression patterns of duplicate gene pairs evolve independently and at vastly different rates. Implications of this finding for the neutralist-selectionist debate are discussed. PMID- 10823905 TI - Operons in Escherichia coli: genomic analyses and predictions. AB - The rich knowledge of operon organization in Escherichia coli, together with the completed chromosomal sequence of this bacterium, enabled us to perform an analysis of distances between genes and of functional relationships of adjacent genes in the same operon, as opposed to adjacent genes in different transcription units. We measured and demonstrated the expected tendencies of genes within operons to have much shorter intergenic distances than genes at the borders of transcription units. A clear peak at short distances between genes in the same operon contrasts with a flat frequency distribution of genes at the borders of transcription units. Also, genes in the same operon tend to have the same physiological functional class. The results of these analyses were used to implement a method to predict the genomic organization of genes into transcription units. The method has a maximum accuracy of 88% correct identification of pairs of adjacent genes to be in an operon, or at the borders of transcription units, and correctly identifies around 75% of the known transcription units when used to predict the transcription unit organization of the E. coli genome. Based on the frequency distance distributions, we estimated a total of 630 to 700 operons in E. coli. This step opens the possibility of predicting operon organization in other bacteria whose genome sequences have been finished. PMID- 10823906 TI - Use of double-stranded RNA interference in Drosophila cell lines to dissect signal transduction pathways. AB - We demonstrate the efficacy of double-stranded RNA-mediated interference (RNAi) of gene expression in generating "knock-out" phenotypes for specific proteins in several Drosophila cell lines. We prove the applicability of this technique for studying signaling cascades by dissecting the well-characterized insulin signal transduction pathway. Specifically, we demonstrate that inhibiting the expression of the DSOR1 (mitogen-activated protein kinase kinase, MAPKK) prevents the activation of the downstream ERK-A (MAPK). In contrast, blocking ERK-A expression results in increased activation of DSOR1. We also show that Drosophila AKT (DAKT) activation depends on the insulin receptor substrate, CHICO (IRS1-4). Finally, we demonstrate that blocking the expression of Drosophila PTEN results in the activation of DAKT. In all cases, the interference of the biochemical cascade by RNAi is consistent with the known steps in the pathway. We extend this powerful technique to study two proteins, DSH3PX1 and Drosophila ACK (DACK). DSH3PX1 is an SH3, phox homology domain-containing protein, and DACK is homologous to the mammalian activated Cdc42 tyrosine kinase, ACK. Using RNAi, we demonstrate that DACK is upstream of DSH3PX1 phosphorylation, making DSH3PX1 an identified downstream target/substrate of ACK-like tyrosine kinases. These experiments highlight the usefulness of RNAi in dissecting complex biochemical signaling cascades and provide a highly effective method for determining the function of the identified genes arising from the Drosophila genome sequencing project. PMID- 10823907 TI - The nonhomologous end-joining pathway of DNA repair is required for genomic stability and the suppression of translocations. AB - We have used spectral karyotyping to assess potential roles of three different components of the nonhomologous DNA end-joining pathway in the maintenance of genomic stability in mouse embryonic fibroblasts (MEFs). MEFs homozygous for mutations that inactivate either DNA ligase IV (Lig4) or Ku70 display dramatic genomic instability, even in the absence of exogenous DNA damaging agents. These aberrant events range from chromosomal fragmentation to nonreciprocal translocations that can involve several chromosomes. DNA-dependent protein kinase catalytic subunit deficiency also promotes genome instability. Deficiency for the p53 cell cycle checkpoint protein has little effect on spontaneous levels of chromosomal instability in Lig4-deficient fibroblasts. However, in the context of ionizing radiation treatment, p53 deficiency allowed visualization of massive acute chromosomal destruction in Lig4-deficient MEFs, which in surviving cells manifested as frequent nonreciprocal translocations. We conclude that nonhomologous DNA end-joining plays a crucial role as a caretaker of the mammalian genome, and that an alternative repair pathway exists that often leads to nonreciprocal translocations. PMID- 10823908 TI - Ectopic scute induces Drosophila ommatidia development without R8 founder photoreceptors. AB - During development of the Drosophila peripheral nervous system, different proneural genes encoding basic helix-loop-helix transcription factors are required for different sensory organs to form. atonal (ato) is the proneural gene required for chordotonal organs and R8 photoreceptors, whereas the achaete-scute complex contains proneural genes for external sensory organs such as the macrochaetae, large sensory bristles. Whereas ectopic ato expression induces chordotonal organ formation, ectopic scute expression produces external sensory organs but not chordotonal organs in the wing. Proneural genes thus appear to specify the sensory organ type. In the ommatidium, or unit eye, R8 is the first photoreceptor to form and appears to recruit other photoreceptors and support cells. In the atonal(1) (ato(1)) mutant, R8 photoreceptors fail to form, thereby resulting in the complete absence of ommatidia. To our surprise, we found that ectopic scute expression in the ato(1) mutant induces the formation of ommatidia, which occasionally sprout ectopic macrochaetae. Remarkably, many scute-induced ommatidia lack R8 although they contain outer photoreceptors. PMID- 10823909 TI - Shear stress-induced apoptosis of adherent neutrophils: a mechanism for persistence of cardiovascular device infections. AB - The mechanisms underlying problematic cardiovascular device-associated infections are not understood. Because the outcome of the acute response to infection is largely dependent on the function of neutrophils, the persistence of these infections suggests that neutrophil function may be compromised because of cellular responses to shear stress. A rotating disk system was used to generate physiologically relevant shear stress levels (0-18 dynes/cm(2); 1 dyne = 10 microN) at the surface of a polyetherurethane urea film. We demonstrate that shear stress diminishes phagocytic ability in neutrophils adherent to a cardiovascular device material, and causes morphological and biochemical alterations that are consistent with those described for apoptosis. Complete neutrophil apoptosis occurred at shear stress levels above 6 dynes/cm(2) after only 1 h. Morphologically, these cells displayed irreversible cytoplasmic and nuclear condensation while maintaining intact membranes. Analysis of neutrophil area and filamentous actin content demonstrated concomitant decreases in both cell area and actin content with increasing levels of shear stress. Neutrophil phagocytosis of adherent bacteria decreased with increasing shear stress. Biochemical alterations included membrane phosphatidylserine exposure and DNA fragmentation, as evaluated by in situ annexin V and terminal deoxynucleotidyltransferase-mediated dUTP end labeling (TUNEL) assays, respectively. The potency of the shear-stress effect was emphasized by comparative inductive studies with adherent neutrophils under static conditions. The combination of tumor necrosis factor-alpha and cycloheximide was ineffective in inducing >21% apoptosis after 3 h. These findings suggest a mechanism through which shear stress plays an important role in the development of bacterial infections at the sites of cardiovascular device implantation. PMID- 10823910 TI - Modeling RNA folding paths with pseudoknots: application to hepatitis delta virus ribozyme. AB - A quantitative understanding of nucleic acid hybridization is essential to many aspects of biotechnology, such as DNA microarrays, as well as to the structure and folding kinetics of RNA. However, predictions of nucleic acid secondary structures have long been impeded by the presence of helices interior to loops, so-called pseudoknots, which impose complex three-dimensional conformational constraints. In this paper we compute the pseudoknot free energies analytically in terms of known standard parameters, and we show how the results can be included in a kinetic Monte Carlo code to follow the succession of secondary structures during quenched or sequential folding. For the hepatitis delta virus ribozyme, we predict several nonnative stems on the folding path, characterize a kinetically trapped state, interpret several experimentally characterized mutations in terms of the folding path, and suggest how hybridization with other parts of the genome inactivates the newly formed ribozyme. PMID- 10823911 TI - Mutations in the molybdenum cofactor biosynthetic protein Cnx1G from Arabidopsis thaliana define functions for molybdopterin binding, molybdenum insertion, and molybdenum cofactor stabilization. AB - The molybdenum cofactor (Moco), a highly conserved pterin compound coordinating molybdenum (Mo), is required for the enzymatic activities of molybdoenzymes. In all organisms studied so far Moco is synthesized by a unique and evolutionary old multistep pathway that requires the activities of at least six gene products. In eukaryotes, the last step of Moco synthesis, i.e., transfer and insertion of Mo into molybdopterin (MPT), is catalyzed by the two-domain proteins Cnx1 in plants and gephyrin in mammals. Both domains (E and G) of these proteins are able to bind MPT in vitro. Here, we show the identification and mutational dissection of functionally important regions within the Cnx1 G domain that are essential for MPT binding, the conversion of MPT to Moco, and Moco stabilization. By functional screening for mutants in the Cnx1 G domain that are no longer able to complement Escherichia coli mogA mutants, we found two classes of mutations in highly conserved amino acid residues. (i) The first class affects in vitro binding of MPT to the protein and the stabilization of Moco, the product of the G domain. (ii) The second class is represented by two independent mutations in the aspartate 515 position that is not affected in MPT binding and Moco stabilization; rather the conversion of MPT to Moco by using bound MPT and a yet unknown form of Mo is completely abolished. The results presented here provide biochemical evidence for a purified Cnx1 G domain catalyzing the insertion of Mo into MPT. PMID- 10823912 TI - Genome evolution of wild barley (Hordeum spontaneum) by BARE-1 retrotransposon dynamics in response to sharp microclimatic divergence. AB - The replicative spread of retrotransposons in the genome creates new insertional polymorphisms, increasing retrotransposon numbers and potentially both their share of the genome and genome size. The BARE-1 retrotransposon constitutes a major, dispersed, active component of Hordeum genomes, and BARE-1 number is positively correlated with genome size. We have examined genome size and BARE-1 insertion patterns and number in wild barley, Hordeum spontaneum, in Evolution Canyon, Lower Nahal Oren, Mount Carmel, Israel, along a transect presenting sharply differing microclimates. BARE-1 has been sufficiently active for its insertional pattern to resolve individuals in a way consonant with their ecogeographical distribution in the canyon and to distinguish them from provenances outside the canyon. On both slopes, but especially on the drier south facing slope, a simultaneous increase in the BARE-1 copy number and a decrease in the relative number lost through recombination, as measured by the abundance of solo long terminal repeats, appear to have driven the BARE-1 share of the genome upward with the height and dryness of the slope. The lower recombinational loss would favor maintenance of more full-length copies, enhancing the ability of the BARE-1 family to contribute to genome size growth. These local data are consistent with regional trends for BARE-1 in H. spontaneum across Israel and therefore may reflect adaptive selection for increasing genome size through retrotransposon activity. PMID- 10823913 TI - Atomic force microscopy reveals the mechanical design of a modular protein. AB - Tandem modular proteins underlie the elasticity of natural adhesives, cell adhesion proteins, and muscle proteins. The fundamental unit of elastic proteins is their individually folded modules. Here, we use protein engineering to construct multimodular proteins composed of Ig modules of different mechanical strength. We examine the mechanical properties of the resulting tandem modular proteins by using single protein atomic force microscopy. We show that by combining modules of known mechanical strength, we can generate proteins with novel elastic properties. Our experiments reveal the simple mechanical design of modular proteins and open the way for the engineering of elastic proteins with defined mechanical properties, which can be used in tissue and fiber engineering. PMID- 10823914 TI - Enhanced insulin secretion and improved glucose tolerance in mice lacking CD26. AB - A subset of prolyl oligopeptidases, including dipeptidyl-peptidase IV (DPP IV or CD26, EC ), specifically cleave off N-terminal dipeptides from substrates having proline or alanine in amino acid position 2. This enzyme activity has been implicated in the regulation of the biological activity of multiple hormones and chemokines, including the insulinotropic peptides glucagon-like peptide 1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP). Targeted inactivation of the CD26 gene yielded healthy mice that have normal blood glucose levels in the fasted state, but reduced glycemic excursion after a glucose challenge. Levels of glucose-stimulated circulating insulin and the intact insulinotropic form of GLP 1 are increased in CD26(-/-) mice. A pharmacological inhibitor of DPP IV enzymatic activity improved glucose tolerance in wild-type, but not in CD26(-/-), mice. This inhibitor also improved glucose tolerance in GLP-1 receptor(-/-) mice, indicating that CD26 contributes to blood glucose regulation by controlling the activity of GLP-1 as well as additional substrates. These data reveal a critical role for CD26 in physiological glucose homeostasis, and establish it as a potential target for therapy in type II diabetes. PMID- 10823915 TI - Structural basis for the interaction of the fluorescence probe 8-anilino-1 naphthalene sulfonate (ANS) with the antibiotic target MurA. AB - The extrinsic fluorescence dye 8-anilino-1-naphthalene sulfonate (ANS) is widely used for probing conformational changes in proteins, yet no detailed structure of ANS bound to any protein has been reported so far. ANS has been successfully used to monitor the induced-fit mechanism of MurA [UDPGlcNAc enolpyruvyltransferase (EC )], an essential enzyme for bacterial cell wall biosynthesis. We have solved the crystal structure of the ANS small middle dotMurA complex at 1.7-A resolution. ANS binds at an originally solvent-exposed region near Pro-112 and induces a major restructuring of the loop Pro-112-Pro-121, such that a specific binding site emerges. The fluorescence probe is sandwiched between the strictly conserved residues Arg-91, Pro-112, and Gly-113. Substrate binding to MurA is accompanied by large movements especially of the loop and Arg-91, which explains why ANS is an excellent sensor of conformational changes during catalysis of this pharmaceutically important enzyme. PMID- 10823916 TI - Assessment of mitochondrial energy coupling in vivo by 13C/31P NMR. AB - The recently cloned uncoupling protein homolog UCP3 is expressed primarily in muscle and therefore may play a significant role in the regulation of energy expenditure and body weight. However, investigation into the regulation of uncoupling protein has been hampered by the inability to assess its activity in vivo. In this report, we demonstrate the use of a noninvasive NMR technique to assess mitochondrial energy uncoupling in skeletal muscle of awake rats by combining (13)C NMR to measure rates of mitochondrial substrate oxidation with (31)P NMR to assess unidirectional ATP synthesis flux. These combined (31)P/(13)C NMR measurements were performed in control, 10-day triiodo-l-thyronine (T(3)) treated (model of increased UCP3 expression), and acute 2,4-dinitrophenol (DNP) treated (protonophore and mitochondrial uncoupler) rats. UCP3 mRNA and protein levels increased 8.1-fold (+/- 1.1) and 2.8-fold (+/- 0.8), respectively, in the T(3)-treated vs. control rat gastrocnemius muscle. (13)C NMR measurements of tricarboxylic acid cycle flux as an index of mitochondrial substrate oxidation were 61 +/- 21, 148 +/- 25, and 310 +/- 48 nmol/g per min in the control, T(3), and DNP groups, respectively. (31)P NMR saturation transfer measurements of unidirectional ATP synthesis flux were 83 +/- 14, 84 +/- 14, and 73 +/- 7 nmol/g per s in the control, T(3), and DNP groups, respectively. Together, these flux measurements, when normalized to the control group, suggest that acute administration of DNP (mitochondrial uncoupler) and chronic administration of T(3) decrease energy coupling by approximately 80% and approximately 60%, respectively, and that the latter treatment correlates with an increase in UCP3 mRNA and protein expression. This NMR approach could prove useful for exploring the regulation of uncoupling protein activity in vivo and elucidating its role in energy metabolism and obesity. PMID- 10823917 TI - Docosahexaenoic acid-containing phospholipid molecular species in brains of vertebrates. AB - The fatty acid composition of phospholipids and the contents of docosahexaenoic acid (DHA)-containing diacyl phosphatidylcholine and diacyl phosphatidylethanolamine molecular species were determined from brains of five fresh-water fish species from a boreal region adapted to 5 degrees C, five fresh water fish species from a temperate region acclimated to 5 degrees C, five fresh water fish species from a temperate region acclimated to 20 degrees C, and three fresh water fish species from a subtropic region adapted to 25-26 degrees C, as well as six mammalian species and seven bird species. There was little difference in DHA levels of fish brains from the different thermal environments; mammalian and bird brain phospholipids contained a few percentage points less DHA than those of the fish investigated. Molecular species of 22:6/22:6, 22:6/20:5, 22:6/20:4, 16:0/22:6, 18:0/22:6, and 18:1/22:6 were identified from all brain probes, and 16:0/22:6, 18:0/22:6, and 18:1/22:6 were the dominating species. Cold water fish brains were rich in 18:1/22:6 diacyl phosphatidylethanolamine (and, to a lesser degree, in diacyl phosphatidylcholine), and its level decreased with increasing environmental/body temperature. The ratio of 18:0/22:6 to 16:0/22:6 phosphatidylcholine and phosphatidylethanolamine was inversely related to body temperature. Phospholipid vesicles from brains of cold-acclimated fish were more fluid, as assessed by using a 1, 6-diphenyl-1,3,5-hexatriene fluorescent probe, than those from bird brains, but the fluidities were almost equal at the respective body temperatures. It is concluded that the relative amounts of these molecular species and their ratios to each other are the major factors contributing to the maintenance of proper fluidity relationships throughout the evolutionary chain as well as helping to maintain important brain functions such as signal transduction and membrane permeability. PMID- 10823918 TI - Identification of the human cytochrome P450, P450RAI-2, which is predominantly expressed in the adult cerebellum and is responsible for all-trans-retinoic acid metabolism. AB - Retinoids, particularly all-trans-retinoic acid (RA), are potent regulators of cell differentiation, cell proliferation, and apoptosis. The role of all-trans-RA during development and in the maintenance of adult tissues has been well established. The control of all-trans-RA levels in cells and tissues is regulated by the balance between its biosynthesis and its catabolism to inactive metabolites. The cytochrome P450 enzyme P450RAI (herein renamed P450RAI-1) is partially responsible for this inactivation of all-trans-RA. In this report, we describe the identification, molecular cloning, and characterization of a second related enzyme, P450RAI-2, which is also involved in the specific inactivation of all-trans-RA. Transiently transfected P450RAI-2 can convert all-trans-RA to more polar metabolites including 4-oxo-, 4-OH-, and 18-OH-all-trans-RA. Competition experiments with other retinoids suggest that all-trans-RA is the preferred substrate. The high level of expression of P450RAI-2, particularly in the cerebellum and pons of human adult brain, suggests a unique role for this enzyme in the protection of specific tissues from exposure to retinoids. PMID- 10823919 TI - A fusion DNA vaccine that targets antigen-presenting cells increases protection from viral challenge. AB - Improving the immunological potency, particularly the Ab response, is a serious hurdle for the protective efficacy and hence broad application of DNA vaccines. We examined the immunogenicity and protective efficacy of a hemagglutinin-based influenza DNA vaccine that was targeted to antigen-presenting cells (APCs) by fusion to CTLA4. The targeted vaccine was shown to induce an accelerated and increased Ab response (as compared with those receiving the nontargeted control) that was predominated by IgG1 and recognized conformationally dependent viral epitopes. Moreover, mice receiving the APC-targeted DNA vaccine had significantly reduced viral titers (100-fold) after a nonlethal virus challenge. The increased protective efficacy was most likely because of increased Ab responses, as cytotoxic T lymphocyte responses were not enhanced. Targeting was demonstrated by direct binding studies of CTLA4 fusion proteins to the cognate ligand (B7; expressed on APCs in vivo). In addition, a targeted protein was detected at 4 fold higher levels in draining lymph nodes within 2-24 h of administration. Therefore, this study demonstrates that targeting DNA-encoded antigen to APCs results in enhanced immunity and strongly suggests that this approach may be useful in improving the protective efficacy of DNA vaccines. PMID- 10823920 TI - Long-term impacts of the Exxon Valdez oil spill on sea otters, assessed through age-dependent mortality patterns. AB - We use age distributions of sea otters (Enhydra lutris) found dead on beaches of western Prince William Sound, Alaska, between 1976 and 1998 in conjunction with time-varying demographic models to test for lingering effects from the 1989 Exxon Valdez oil spill. Our results show that sea otters in this area had decreased survival rates in the years following the spill and that the effects of the spill on annual survival increased rather than dissipated for older animals. Otters born after the 1989 spill were affected less than those alive in March 1989, but do show continuing negative effects through 1998. Population-wide effects of the spill appear to have slowly dissipated through time, due largely to the loss of cohorts alive during the spill. Our results demonstrate that the difficult-to detect long-term impacts of environmental disasters may still be highly significant and can be rigorously analyzed by using a combination of population data, modeling techniques, and statistical analyses. PMID- 10823921 TI - The steroid receptor coactivator SRC-3 (p/CIP/RAC3/AIB1/ACTR/TRAM-1) is required for normal growth, puberty, female reproductive function, and mammary gland development. AB - Steroid receptor coactivator-3 (SRC-3) is a coactivator of nuclear receptors in the SRC family as assayed in vitro. Here, we show that mouse SRC-3 is expressed in a tissue-specific fashion and distributed mainly in the oocytes, mammary glands, hippocampus, olfactory bulb, smooth muscle, hepatocytes, and vaginal epithelium. Genetic disruption of SRC-3 in mice results in a pleiotropic phenotype showing dwarfism, delayed puberty, reduced female reproductive function, and blunted mammary gland development. Hormonal analysis indicates that SRC-3 plays a role in both the growth hormone regulatory pathway and the production of estrogen, which may explain the observed phenotypes. These results suggest that the physiological role of SRC-3 is different from that of SRC-1 and prove the diversity among coactivator family members. PMID- 10823922 TI - A protein required for prion generation: [URE3] induction requires the Ras regulated Mks1 protein. AB - Infectious proteins (prions) can arise de novo as well as by transmission from another individual. De novo prion generation is believed responsible for most cases of Creutzfeldt-Jakob disease and for initiating the mad cow disease epidemic. However, the cellular components needed for prion generation have not been identified in any system. The [URE3] prion of Saccharomyces cerevisiae is an infectious form of Ure2p, apparently a self-propagating amyloid. We now demonstrate a protein required for de novo prion generation. Mks1p negatively regulates Ure2p and is itself negatively regulated by the presence of ammonia and by the Ras-cAMP pathway. We find that in mks1Delta strains, de novo generation of the [URE3] prion is blocked, although [URE3] introduced from another strain is expressed and propagates stably. Ras2(Val19) increases cAMP production and also blocks [URE3] generation. These results emphasize the distinction between prion generation and propagation, and they show that cellular regulatory mechanisms can critically affect prion generation. PMID- 10823923 TI - The Arabidopsis thaliana salt tolerance gene SOS1 encodes a putative Na+/H+ antiporter. AB - In Arabidopsis thaliana, the SOS1 (Salt Overly Sensitive 1) locus is essential for Na(+) and K(+) homeostasis, and sos1 mutations render plants more sensitive to growth inhibition by high Na(+) and low K(+) environments. SOS1 is cloned and predicted to encode a 127-kDa protein with 12 transmembrane domains in the N terminal part and a long hydrophilic cytoplasmic tail in the C-terminal part. The transmembrane region of SOS1 has significant sequence similarities to plasma membrane Na(+)/H(+) antiporters from bacteria and fungi. Sequence analysis of various sos1 mutant alleles reveals several residues and regions in the transmembrane as well as the tail parts that are critical for SOS1 function in plant salt tolerance. SOS1 gene expression in plants is up-regulated in response to NaCl stress. This up-regulation is abated in sos3 or sos2 mutant plants, suggesting that it is controlled by the SOS3/SOS2 regulatory pathway. PMID- 10823924 TI - Inactivation of the acid labile subunit gene in mice results in mild retardation of postnatal growth despite profound disruptions in the circulating insulin-like growth factor system. AB - Insulin-like growth factors (IGFs) I and II are important regulators of cell proliferation and differentiation. After birth, plasma IGFs, representing mostly liver-derived IGFs, circulate in ternary complexes of 150 kDa consisting of one molecule each of IGF, IGF-binding protein (IGFBP) 3, and an acid labile subunit (ALS). Onset of ALS synthesis after birth is the primary factor driving the formation of ternary complexes. Capture of IGFs by ALS is thought to allow the development of a plasma reservoir without negative effects such as hypoglycemia and cell proliferation. To evaluate the importance of ALS and ternary complexes, we have created mice in which the ALS gene has been inactivated. The mutation was inherited in a Mendelian manner, without any effects on survival rates and birth weights. A growth deficit was observed in null mice after 3 weeks of life and reached 13% by 10 weeks. This modest phenotype was observed despite reductions of 62 and 88% in the concentrations of plasma IGF-I and IGFBP-3, respectively. Increased turnover accounted for these reductions because indices of synthesis in liver and kidney were not decreased. Surprisingly, absence of ALS did not affect glucose and insulin homeostasis. Therefore, ALS is required for postnatal accumulation of IGF-I and IGFBP-3 but, consistent with findings supporting a predominant role for locally produced IGF-I, is not critical for growth. This model should be useful to determine whether presence of ALS is needed for other actions of liver-derived IGF-I and for maintenance of homeostasis in presence of high circulating levels of IGF-II. PMID- 10823925 TI - Unpaired terminal nucleotides and 5' monophosphorylation govern 3' polyadenylation by Escherichia coli poly(A) polymerase I. AB - In bacteria, most mRNAs and certain regulatory RNAs are rapidly turned over, whereas mature tRNA and ribosomal RNA are highly stable. The selective susceptibility of unstable Escherichia coli RNAs to 3' polyadenylation by the pcnB gene product, poly(A) polymerase I (PAP I), in vivo is a key factor in their rapid degradation by 3' to 5' exonucleases. Using highly purified His-tagged recombinant PAP I, we show that differential adenylation of RNA substrates by PAP I occurs in vitro and that this capability resides in PAP I itself rather than in any ancillary protein(s). Surprisingly, the efficiency of 3' polyadenylation is affected by substrate structure at both termini; single-strand segments at either the 5' or 3' end of RNA molecules and monophosphorylation at an unpaired 5' terminus dramatically increase the rate and length of 3' poly(A) tail additions by PAP I. Our results provide a mechanistic basis for the susceptibility of certain RNAs to 3' polyadenylation. They also suggest a model of "programmed" RNA decay in which endonucleolytically generated RNA fragments containing single stranded monophosphorylated 5' termini are targeted for poly(A) addition and further degradation. PMID- 10823926 TI - Down-regulation of iron regulatory protein 1 gene expression by nitric oxide. AB - Iron regulatory protein 1 (IRP1) is an RNA binding protein that posttranscriptionally modulates the expression of mRNAs coding for proteins involved in iron metabolism. It has long been held that its RNA binding activity is regulated posttranslationally by the insertion/extrusion of a 4Fe-4S cluster, without changes in IRP1 levels. However, the question of a possible regulation of the expression of this protein has remained open. In the present study we analyzed the modulation of IRP1 expression in murine macrophages. We showed that activation by IFN-gamma and/or lipopolysaccharide, which induces IRP1 RNA binding activity via nitric oxide (NO), results simultaneously in a reduction in IRP1 protein levels, as determined by Western blot analyses. IRP1 expression decreased time-dependently to about 40% of control levels after 16 h. Down-regulation of IRP1 protein levels was correlated with the amount of NO produced and was partially abolished by the NO synthase (NOS) inhibitor N-monomethyl-l-arginine. No changes in IRP1 levels could be detected in stimulated peritoneal macrophages from NOS2 knockout (NOS2(-/-)) mice, unlike wild-type mice. Converse modulation of IRP1 RNA binding activity and IRP1 levels could be reproduced by exogenous NO and also was observed in nonmacrophage cells cocultured with NO-producing macrophages. We also analyzed IRP1 mRNA levels by Northern blotting and found a decrease in IRP1 mRNA expression after stimulation with IFN-gamma plus lipopolysaccharide, which was abrogated in the presence of N-monomethyl-l arginine. This is evidence that IRP1 is regulated by a physiological stimulus other than posttranslationally. PMID- 10823927 TI - HSV.com: maneuvering the internetworks of viral neuropathogenesis and evasion of the host defense. PMID- 10823928 TI - New paradigm for drug therapies of cardiac fibrillation. PMID- 10823929 TI - In search of gamma-secretase: presenilin at the cutting edge. PMID- 10823930 TI - Ecological character displacement and the study of adaptation. PMID- 10823931 TI - A route to hierarchical materials based on complexes of metallosupramolecular polyelectrolytes and amphiphiles. AB - Anisotropic thin film materials of metallosupramolecular polyelectrolyte amphiphile complexes (denoted PACs) with structures at several length scales were fabricated through a multistep self-assembly process. Metal ion-mediated self assembly of the ditopic ligand 1,4-bis(2,2':6',2"-terpyridine-4'-yl)benzene and electrostatic binding with the amphiphile dihexadecyl phosphate result in a PAC with tailored surface chemical properties, including solubility and surface activity. The PAC forms a stable monolayer at the air-water interface that is readily transferred and oriented on solid supports with the Langmuir-Blodgett technique. The presented strategy unifies colloid and metallosupramolecular chemistry and opens a versatile route to hierarchical materials with tailored structures and functions. PMID- 10823932 TI - A mammalian germ cell-specific RNA-binding protein interacts with ubiquitously expressed proteins involved in splice site selection. AB - RNA-binding motif (RBM) genes are found on all mammalian Y chromosomes and are implicated in spermatogenesis. Within human germ cells, RBM protein shows a similar nuclear distribution to components of the pre-mRNA splicing machinery. To address the function of RBM, we have used protein-protein interaction assays to test for possible physical interactions between these proteins. We find that RBM protein directly interacts with members of the SR family of splicing factors and, in addition, strongly interacts with itself. We have mapped the protein domains responsible for mediating these interactions and expressed the mouse RBM interaction region as a bacterial fusion protein. This fusion protein can pull down several functionally active SR protein species from cell extracts. Depletion and add-back experiments indicate that these SR proteins are the only splicing factors bound by RBM which are required for the splicing of a panel of pre-mRNAs. Our results suggest that RBM protein is an evolutionarily conserved mammalian splicing regulator which operates as a germ cell-specific cofactor for more ubiquitously expressed pre-mRNA splicing activators. PMID- 10823933 TI - Bcl-2 decreases the free Ca2+ concentration within the endoplasmic reticulum. AB - The antiapoptotic protein Bcl-2 localizes not only to mitochondria but also to the endoplasmic reticulum (ER). However, the function of Bcl-2 at the level of the ER is poorly understood. In this study, we have investigated the effects of Bcl-2 expression on Ca(2+) storage and release by the ER. The expression of Bcl-2 decreased the amount of Ca(2+) that could be released from intracellular stores, regardless of the mode of store depletion, the cell type, or the species from which Bcl-2 was derived. Bcl-2 also decreased cellular Ca(2+) store content in the presence of mitochondrial inhibitors, suggesting that its effects were not mediated through mitochondrial Ca(2+) uptake. Direct measurements with ER targeted Ca(2+)-sensitive fluorescent "cameleon" proteins revealed that Bcl-2 decreased the free Ca(2+) concentration within the lumen of the ER, [Ca(2+)](ER). Analysis of the kinetics of Ca(2+) store depletion in response to the Ca(2+) ATPase inhibitor thapsigargin revealed that Bcl-2 increased the permeability of the ER membrane. These results suggest that Bcl-2 decreases the free Ca(2+) concentration within the ER lumen by increasing the Ca(2+) permeability of the ER membrane. The increased ER Ca(2+) permeability conferred by Bcl-2 would be compatible with an ion channel function of Bcl-2 at the level of the ER membrane. PMID- 10823934 TI - Specific interaction of CCR5 amino-terminal domain peptides containing sulfotyrosines with HIV-1 envelope glycoprotein gp120. AB - The HIV-1 envelope glycoprotein gp120 interacts consecutively with CD4 and the CCR5 coreceptor to mediate the entry of certain HIV-1 strains into target cells. Acidic residues and sulfotyrosines in the amino-terminal domain (Nt) of CCR5 are crucial for viral fusion and entry. We tested the binding of a panel of CCR5 Nt peptides to different soluble gp120/CD4 complexes and anti-CCR5 mAbs. The tyrosine residues in the peptides were sulfated, phosphorylated, or unmodified. None of the gp120/CD4 complexes associated with peptides containing unmodified or phosphorylated tyrosines. The gp120/CD4 complexes containing envelope glycoproteins from isolates that use CCR5 as a coreceptor associated with Nt peptides containing sulfotyrosines but not with peptides containing sulfotyrosines in scrambled Nt sequences. Finally, only peptides containing sulfotyrosines inhibited the entry of an R5 isolate. Our data show that proper posttranslational modification of the CCR5 Nt is required for gp120 binding and viral entry. More importantly, the Nt domain determines the specificity of the interaction between CCR5 and gp120s from isolates that use this coreceptor. PMID- 10823935 TI - A 12(S)-hydroxyeicosatetraenoic acid receptor interacts with steroid receptor coactivator-1. AB - Lewis lung carcinoma cells contain specific high-affinity binding sites for the eicosanoid 12(S)-hydroxy-5,8,10,14-eicosatetraenoic acid [12(S)-HETE]. These binding sites have a cytosolic/nuclear localization and contain the heat shock proteins hsp70 and hsp90 as components of a high molecular weight cytosolic binding complex. The ligand binding subunit of this complex is a protein with an apparent molecular mass of approximately 50 kDa as judged by gel permeation chromatography. In this report, we present data showing that the 50-kDa 12(S) HETE binding protein interacts as a homodimer with steroid receptor coactivator-1 (SRC-1) in the presence of 12(S)-HETE. Two putative interaction domains were mapped. One of these (amino acids 701-781) was within the nuclear receptor interaction domain in SRC-1 required for binding of various steroid and thyroid hormone receptors. It contains the most C-terminal of the three copies of LXXLL motif present in the nuclear receptor interaction domain. The second interaction domain was present in the N-terminal part of SRC-1 (amino acids 1-221). This region has two LXXLL motifs, one does not bind and the other binds only weakly to steroid and thyroid hormone receptors. Glutathione S-transferase (GST) pulldown experiments and far Western analyses demonstrated that the N-terminal region of SRC-1 (amino acids 1-212) alone does not bind the 50-kDa 12(S)-HETE binding protein, whereas GST/DeltaSRC-1(1-1138) ligand-dependently pulled down a protein of approximately 50 kDa in size. Our results suggest that the 50-kDa 12(S)-HETE binding protein is a receptor that may signal through interaction with a nuclear receptor coactivator protein. PMID- 10823936 TI - Structural diversity of self-cleaving ribozymes. AB - In vitro selection was used to isolate Mg(2+)-dependent self-cleaving ribozymes from random sequence. Characterization of representative clones revealed the emergence of at least 12 classes of ribozymes that adopt distinct secondary structure motifs. Only one class corresponds to a previously known structural motif, that of the naturally occurring hammerhead ribozyme. Each ribozyme promotes self-cleavage via an internal phosphoester transfer reaction involving the adjacent 2'-hydroxyl group with a chemical rate enhancement of between 10(3)- and 10(6)-fold greater than the corresponding uncatalyzed rate. These findings indicate that RNA can form a multitude of secondary and tertiary structures that promote cleavage by internal phosphoester transfer. Upon further in vitro selection, a class I ribozyme that adopts an "X motif" structure dominates over all other ribozymes in the population. Thus, self-cleaving RNAs isolated by in vitro selection from random-sequence populations can rival the catalytic efficiency of natural ribozymes. PMID- 10823937 TI - The major transition state in folding need not involve the immobilization of side chains. AB - During protein folding in which few, if any, definable kinetic intermediates are observable, the nature of the transition state is central to understanding the course of the reaction. Current experimental data does not distinguish the relative contributions of side chain immobilization and dehydration phenomena to the major rate-limiting transition state whereas this distinction is central to theoretical models that attempt to simulate the behavior of proteins during folding. Renaturation of the small proteinase inhibitor cystatin under oxidizing versus reducing conditions is the first experimental case in which these processes can be studied independently. Using this example, we show that sidechain immobilization occurs downstream of the major folding transition state. A consequence of this is the existence of states with disordered side chains, which are distinct from kinetic protein folding intermediates and which lie within the folded state free energy well. PMID- 10823938 TI - Internal packing of helical membrane proteins. AB - Helix packing is important in the folding, stability, and association of membrane proteins. Packing analysis of the helical portions of 7 integral membrane proteins and 37 soluble proteins show that the helices in membrane proteins have higher packing values (0.431) than in soluble proteins (0.405). The highest packing values in integral membrane proteins originate from small hydrophobic (G and A) and small hydroxyl-containing (S and T) amino acids, whereas in soluble proteins large hydrophobic and aromatic residues have the highest packing values. The highest packing values for membrane proteins are found in the transmembrane helix-helix interfaces. Glycine and alanine have the highest occurrence among the buried amino acids in membrane proteins, whereas leucine and alanine are the most common buried residue in soluble proteins. These observations are consistent with a shorter axial separation between helices in membrane proteins. The tight helix packing revealed in this analysis contributes to membrane protein stability and likely compensates for the lack of the hydrophobic effect as a driving force for helix-helix association in membranes. PMID- 10823939 TI - Scaffold proteins may biphasically affect the levels of mitogen-activated protein kinase signaling and reduce its threshold properties. AB - In addition to preventing crosstalk among related signaling pathways, scaffold proteins might facilitate signal transduction by preforming multimolecular complexes that can be rapidly activated by incoming signal. In many cases, such as mitogen-activated protein kinase (MAPK) cascades, scaffold proteins are necessary for full activation of a signaling pathway. To date, however, no detailed biochemical model of scaffold action has been suggested. Here we describe a quantitative computer model of MAPK cascade with a generic scaffold protein. Analysis of this model reveals that formation of scaffold-kinase complexes can be used effectively to regulate the specificity, efficiency, and amplitude of signal propagation. In particular, for any generic scaffold there exists a concentration value optimal for signal amplitude. The location of the optimum is determined by the concentrations of the kinases rather than their binding constants and in this way is scaffold independent. This effect and the alteration of threshold properties of the signal propagation at high scaffold concentrations might alter local signaling properties at different subcellular compartments. Different scaffold levels and types might then confer specialized properties to tune evolutionarily conserved signaling modules to specific cellular contexts. PMID- 10823940 TI - Structure of a two-domain chitotriosidase from Serratia marcescens at 1.9-A resolution. AB - In this paper, we describe the structure of chitinase B from Serratia marcescens, which consists of a catalytic domain with a TIM-barrel fold and a 49-residue C terminal chitin-binding domain. This chitinase is the first structure of a bacterial exochitinase, and it represents one of only a few examples of a glycosyl hydrolase structure having interacting catalytic and substrate-binding domains. The chitin-binding domain has exposed aromatic residues that contribute to a 55-A long continuous aromatic stretch extending into the active site. Binding of chitin oligomers is blocked beyond the -3 subsite, which explains why the enzyme has chitotriosidase activity and degrades the chitin chain from the nonreducing end. Comparison of the chitinase B structure with that of chitinase A explains why these enzymes act synergistically in the degradation of chitin. PMID- 10823942 TI - Cloning and characterization of the mammalian brain-specific, Mg2+-dependent neutral sphingomyelinase. AB - The enzymatic breakdown of sphingomyelin by sphingomyelinases is considered the major source of the second messenger ceramide. Studies on the contribution of the various described acidic and neutral sphingomyelinases to the signaling pool of ceramide have been hampered by the lack of molecular data on the neutral sphingomyelinases (nSMases). We recently identified a mammalian nSMase, an integral membrane protein with remote similarity to bacterial sphingomyelinases. However, its ubiquitous expression pattern is in contrast to previous findings that sphingomyelinase activity is found mainly in brain tissues. By using an improved database search method, combined with phylogenetic analysis, we identified a second mammalian nSMase (nSMase2) with predominant expression in the brain. The sphingomyelinase activity of nSMase2 has a neutral pH optimum, depends on Mg(2+) ions, and is activated by unsaturated fatty acids and phosphatidylserine. Immunofluorescence reveals a neuron-specific punctate perinuclear staining, which colocalizes with a Golgi marker in a number of cell lines. The likely identity of nSMase2 with cca1, a rat protein involved in contact inhibition of 3Y1 fibroblasts, suggests a role for this enzyme in cell cycle arrest. Both mammalian nSMases are members of a superfamily of Mg(2+) dependent phosphohydrolases, which also contains nucleases, inositol phosphatases, and bacterial toxins. PMID- 10823941 TI - Endoplasmic reticulum retention is a common defect associated with tyrosinase negative albinism. AB - Tyrosinase is a melanocyte-specific enzyme critical for the synthesis of melanin, a process normally restricted to a post-Golgi compartment termed the melanosome. Loss-of-function mutations in tyrosinase are the cause of oculocutaneous albinism, demonstrating the importance of the enzyme in pigmentation. In the present study, we explored the possibility that trafficking of albino tyrosinase from the endoplasmic reticulum (ER) to the Golgi apparatus and beyond is disrupted. Toward this end, we analyzed the common albino mouse mutation Tyr(C85S), the frequent human albino substitution TYR(T373K), and the temperature sensitive tyrosinase TYR(R402Q)/Tyr(H402A) found in humans and mice, respectively. Intracellular localization was monitored in albino melanocytes carrying the native mutation, as well as in melanocytes ectopically expressing green fluorescent protein-tagged tyrosinase. Enzymatic characterization of complex glycans and immunofluorescence colocalization with organelle-specific resident proteins established that all four mutations produced defective proteins that were retained in the ER. TYR(R402Q)/Tyr(H402A) Golgi processing and transport to melanosomes were promoted at the permissive temperature of 32 degrees C, but not at the nonpermissive 37 degrees C temperature. Furthermore, evidence of protein misfolding was demonstrated by the prolonged association of tyrosinase mutants with calnexin and calreticulin, known ER chaperones that play a key role in the quality-control processes of the secretory pathway. From these results we concluded that albinism, at least in part, is an ER retention disease. PMID- 10823943 TI - Nuclear clusterin/XIP8, an x-ray-induced Ku70-binding protein that signals cell death. AB - Clusterin [CLU, a.k.a. TRPM-2, SGP-2, or ionizing radiation (IR)-induced protein 8 (XIP8)] was implicated in apoptosis, tissue injury, and aging. Its function remains elusive. We reisolated CLU/XIP8 by yeast two-hybrid analyses using as bait the DNA double-strand break repair protein Ku70. We show that a delayed (2-3 days), low-dose (0.02-10 Gy) IR-inducible nuclear CLU/XIP8 protein coimmunoprecipitated and colocalized (by confocal microscopy) in vivo with Ku70/Ku80, a DNA damage sensor and key double-strand break repair protein, in human MCF-7:WS8 breast cancer cells. Overexpression of nuclear CLU/XIP8 or its minimal Ku70 binding domain (120 aa of CLU/XIP8 C terminus) in nonirradiated MCF 7:WS8 cells dramatically reduced cell growth and colony-forming ability concomitant with increased G(1) cell cycle checkpoint arrest and increased cell death. Enhanced expression and accumulation of nuclear CLU/XIP8-Ku70/Ku80 complexes appears to be an important cell death signal after IR exposure. PMID- 10823944 TI - Identification of a human centrosomal calmodulin-binding protein that shares homology with pericentrin. AB - Eukaryotic chromosome segregation depends on the mitotic spindle apparatus, a bipolar array of microtubules nucleated from centrosomes. Centrosomal microtubule nucleation requires attachment of gamma-tubulin ring complexes to a salt insoluble centrosomal core, but the factor(s) underlying this attachment remains unknown. In budding yeast, this attachment is provided by the coiled-coil protein Spc110p, which links the yeast gamma-tubulin complex to the core of the yeast centrosome. Here, we show that the large coiled-coil protein kendrin is a human orthologue of Spc110p. We identified kendrin by its C-terminal calmodulin-binding site, which shares homology with the Spc110p calmodulin-binding site. Kendrin localizes specifically to centrosomes throughout the cell cycle. N-terminal regions of kendrin share significant sequence homology with pericentrin, a previously identified murine centrosome component known to interact with gamma tubulin. In mitotic human breast carcinoma cells containing abundant centrosome like structures, kendrin is found only at centrosomes associated with spindle microtubules. PMID- 10823945 TI - Human vascular endothelial cells contain membrane binding sites for estradiol, which mediate rapid intracellular signaling. AB - Estrogen induces both rapid and delayed effects on the cardiovascular system. The early effects take place within minutes (e.g., changes in vasomotor tone) and are mediated through rapid intracellular signaling pathways; whereas the delayed effects (e.g., remodeling or lipid alterations) require hours to days to occur and require transcriptional effects with subsequent modulation of protein expression. To study the acute effects of 17beta-estradiol (E2) treatment on vascular function, we have investigated the rapid (on the order of minutes) effects of E2 treatment on intracellular signaling in human endothelial cells (EC). Our previous data have shown that E2 induces rapid release of NO from and activation of guanylate cyclase in human EC. In this study, we demonstrate that E2 also activates mitogen-activated protein kinase (extracellular signal-related kinase) signaling within minutes in EC. We hypothesized that this effect might be mediated by estrogen receptors (ER) localized to the cell surface. Our data show that membrane-impermeant forms of E2 also activate EC mitogen-activated protein kinase as well as stimulate cGMP production and NO release. The ER antagonist ICI 182,780 blocks this effect. Using confocal microscopy and flow cytometric analysis, we demonstrate that EC contain surface binding sites for E2, detectable by cell-impermeant ligand binding and equally with an anti-ERalpha antibody. Immunoreactive bands of 66 and 45 kDa are detectable with an anti-ERalpha mAb in human EC, and their individual presence correlates functionally with E2 stimulated genomic and rapid nongenomic responses, respectively. Membrane ERs may provide key molecular switches in these novel, rapid signaling pathways induced by E2 in EC. PMID- 10823946 TI - Estrogen receptor (ER) beta, a modulator of ERalpha in the uterus. AB - Many of the effects of estrogens on the uterus are mediated by ERalpha, the predominant ER in the mature organ. Because of the poor reproductive capacity of ERbeta knockout (BERKO) female mice (small litter size, multiple-resorbed fetuses), the role of uterine ERbeta was explored. In the immature uterus, ERalpha and ERbeta are expressed at comparable levels in the epithelium and stroma, and 17beta-estradiol (E(2)) treatment decreases ERbeta in the stroma. The immature uterus of untreated BERKO mice exhibits elevated levels of progesterone receptor (PR) and the proliferation-associated protein, Ki-67. It also exhibits exaggerated responsiveness to E(2), as indicated by enlargement of the lumen, increase in volume and protein content of uterine secretion, induction of the luminal epithelial secretory protein, complement C3, and its regulatory cytokine IL-1beta, and induction of vascular endothelial growth factor and insulin-like growth factor 1 but not its receptor. As expected, E(2) increased PR in the stroma and decreased it in the luminal epithelium of wild-type mice. In the BERKO uterus, E(2) induced PR in the stroma but did not down-regulate it in the epithelium. Increased cell proliferation and exaggerated response to E(2) in BERKO suggest that ERbeta plays a role in modulation of the effects of ERalpha and in addition (or as a consequence of this) has an antiproliferative function in the immature uterus. PMID- 10823947 TI - Reduced X-linked nucleotide polymorphism in Drosophila simulans. AB - Population genetic theory predicts that selectively driven changes of allele frequency for both beneficial and deleterious mutants reduce polymorphism at tightly linked sites. All else being equal, these reductions in polymorphism are expected to be greater when recombination rates are lower. Therefore, the empirical observation of a positive correlation between recombination rates and amounts of DNA polymorphism across the Drosophila melanogaster genome can be explained by two very different types of natural selection. Here, we evaluate alternative models of effects of selection on linked sites by comparison of X linked and autosomal variation. We present polymorphism data from 40 genes distributed across chromosome arms X and 3R of Drosophila simulans, a sibling species of D. melanogaster. We find significantly less silent polymorphism in D. simulans on the X chromosome than on 3R, but no difference between arms for silent divergence between species. This pattern is incompatible with predictions from theoretical studies on the effect of negative selection on linked sites. We propose that some form of positive selection having greater effects on sex chromosomes than on autosomes is the better explanation for the D. simulans data. PMID- 10823948 TI - Cholesterol-dependent clustering of IL-2Ralpha and its colocalization with HLA and CD48 on T lymphoma cells suggest their functional association with lipid rafts. AB - Immunogold staining and electron microscopy show that IL-2 receptor alpha subunits exhibit nonrandom surface distribution on human T lymphoma cells. Analysis of interparticle distances reveals that this clustering on the scale of a few hundred nanometers is independent of the presence of IL-2 and of the expression of the IL-2R beta-subunit. Clustering of IL-2Ralpha is confirmed by confocal microscopy, yielding the same average cluster size, approximately 600 800 nm, as electron microscopy. HLA class I and II and CD48 molecules also form clusters of the same size. Disruption of cholesterol-rich lipid rafts with filipin or depletion of membrane cholesterol with methyl-beta-cyclodextrin results in the blurring of cluster boundaries and an apparent dispersion of clusters for all four proteins. Interestingly, the transferrin receptor, which is thought to be located outside lipid rafts, exhibits clusters that are only 300 nm in size and are less affected by modifying the membrane cholesterol content. Furthermore, transferrin receptor clusters hardly colocalize with IL-2Ralpha, HLA, and CD48 molecules (crosscorrelation coefficient is 0.05), whereas IL 2Ralpha colocalizes with both HLA and CD48 (crosscorrelation coefficient is between 0.37 and 0.46). This coclustering is confirmed by electron microscopy. The submicron clusters of IL-2Ralpha chains and their coclustering with HLA and CD48, presumably associated with lipid rafts, could underlie the efficiency of signaling in lymphoid cells. PMID- 10823949 TI - Inflammation in the developing human intestine: A possible pathophysiologic contribution to necrotizing enterocolitis. AB - Necrotizing enterocolitis (NEC), a major cause of morbidity and mortality in premature infants, occurs after the introduction of oral feedings in conjunction with initial bacterial colonization of the gut and is hypothesized to be due to an immature (inappropriate) enterocyte response to bacterial stimuli. To test this hypothesis, we compared the enterocyte IL-8 response to inflammatory stimuli [lipopolysaccharide (LPS) and IL-1beta] in immature vs. mature human small intestine. Initial in vitro studies comparing confluent Caco-2 cells, a model for mature human enterocytes, with a primary human fetal intestinal cell line (H4 cells) demonstrated that after inflammatory stimulation fetal cells secreted more IL-8 (LPS, 8-fold; IL-1beta, 20-fold) than Caco-2 cells. IL-8 mRNA activity in fetal compared to Caco-2 cells was proportionately increased by the same magnitude with both stimuli. To validate the in vitro observations, small intestinal organ cultures from fetuses vs. older children were exposed to LPS and IL-1beta. Again in human organ cultures from fetuses compared to older children, IL-8 secretion was greater (LPS, 2.5-fold; IL-1beta, 200-fold) and mRNA activity after stimulation was comparably higher, suggesting that increased transcription of the IL-8 gene may account for the excessive response. Using immunohistochemical staining to identify the cellular source of IL-8, activity was noted predominantly in villous and crypt epithelium but also in a few immunoresponsive lymphoid cells. The observation that immature human enterocytes react with excessive pro-inflammatory cytokine production after inflammatory stimulation may help in part explain why prematures exposed to initial colonizing bacteria develop necrotizing enterocolitis. PMID- 10823950 TI - Cancer incidences in Europe related to mortalities, and ethnohistoric, genetic, and geographic distances. AB - We have previously shown that geographic differences in cancer mortalities in Europe are related to (in order of importance): geographic distances (reflecting environmental differences), ethnohistoric distances (encompassing cultural and genetic attributes), and genetic distances of the populations in the areas studied. In this study, we analyzed the relations of the same three factors to European incidences of 45 male and 47 female cancers. Differences in cancer incidences are correlated moderately, first with geographic distances, and then with genetic distances, but not at all with ethnohistoric distances. Comparing these findings to the earlier ones for cancer mortalities, we note the reversal in the importance of ethnohistory and genetics, and the generally lower correlations of incidence differences with the three putatively causal distance matrices. A path diagram combining both studies demonstrates the lack of cultural carcinogenic effects, but suggests cultural influences on procedures such as the registration of deaths in different political entities. Additionally, the relatively large correlation between ethnohistoric distances and mortality differences is caused by common factors behind the correlation of ethnohistoric and geographic distances. Geographic proximity results in similar ethnohistories. The direct effects of genetic distances are negligible and only their common effects with geographic distances play a role, accounting for the weak to negligible influence of genetics on incidence and mortality differences. Apparently, the genetic systems available to us do not substantially affect cancer incidence or mortality. We present indirect evidence that international differences in the quality of cancer rate data are greater in mortalities than in incidences. PMID- 10823951 TI - Mimicking dominant negative inhibition of prion replication through structure based drug design. AB - Recent progress determining the structure of the host-encoded prion protein (PrP(C)) and the role of auxiliary molecules in prion replication permits a more rational approach in the development of therapeutic interventions. Our objective is to identify a new class of lead compounds that mimic the dominant negative PrP(C) mutants, which inhibit an abnormal isoform (PrP(Sc)) formation. A computational search was conducted on the Available Chemicals Directory for molecules that mimic both the spatial orientation and basic polymorphism of PrP residues 168, 172, 215, and 219, which confer dominant negative inhibition. The search revealed 1,000 potential candidates that were visually analyzed with respect to the structure of this four-residue epitope on PrP(C). Sixty-three compounds were tested for inhibition of PrP(Sc) formation in scrapie-infected mouse neuroblastoma cells (ScN2a). Two compounds, Cp-60 (2-amino-6-[(2 aminophenyl)thio]-4-(2-furyl)pyridine-3, 5-dicarbonitrile) and Cp-62 (N'1-(?5 [(4, 5-dichloro-1H-imidazol-1-yl)methyl]-2-furyl?carbonyl)-4 methoxybenzene-1 sulfonohydrazide), inhibited PrP(Sc) formation in a dose-dependent manner and demonstrated low levels of toxicity. A substructure search of the Available Chemicals Directory based on Cp-60 identified five related molecules, three of which exhibited activities comparable to Cp-60. Mimicking dominant negative inhibition in the design of drugs that inhibit prion replication may provide a more general approach to developing therapeutics for deleterious protein-protein interactions. PMID- 10823952 TI - Oral administration of a corticotropin-releasing hormone receptor antagonist significantly attenuates behavioral, neuroendocrine, and autonomic responses to stress in primates. AB - We evaluated the effects of the lipophilic nonpeptide corticotropin-releasing hormone (CRH) type 1 receptor antagonist antalarmin on the behavioral, neuroendocrine, and autonomic components of the stress response in adult male rhesus macaques. After oral administration, significant antalarmin concentrations were detected in the systemic circulation and the cerebrospinal fluid by a mass spectrometry-gas chromatography assay developed specifically for this purpose. Pharmacokinetic and dose-response studies suggested that an oral dose of 20 mg/kg was optimal for behavioral and endocrine effects. We then administered this dose in a double-blind, placebo-controlled fashion to monkeys exposed to an intense social stressor: namely, placement of two unfamiliar males in adjacent cages separated only by a transparent Plexiglas screen. Antalarmin significantly inhibited a repertoire of behaviors associated with anxiety and fear such as body tremors, grimacing, teeth gnashing, urination, and defecation. In contrast, antalarmin increased exploratory and sexual behaviors that are normally suppressed during stress. Moreover, antalarmin significantly diminished the increases in cerebrospinal fluid CRH as well as the pituitary-adrenal, sympathetic, and adrenal medullary responses to stress. We conclude that CRH plays a broad role in the physiological responses to psychological stress in primates and that a CRH type 1 receptor antagonist may be of therapeutic value in human psychiatric, reproductive, and cardiovascular disorders associated with CRH system hyperactivity. PMID- 10823953 TI - Mutations in the tuberous sclerosis complex gene TSC2 are a cause of sporadic pulmonary lymphangioleiomyomatosis. AB - Lymphangioleiomyomatosis (LAM) is a progressive and often fatal interstitial lung disease characterized by a diffuse proliferation of abnormal smooth muscle cells in the lungs. LAM is of unusual interest biologically because it affects almost exclusively young women. LAM can occur as an isolated disorder (sporadic LAM) or in association with tuberous sclerosis complex. Renal angiomyolipomas, which are found in most tuberous sclerosis patients, also occur in 60% of sporadic LAM patients. We previously found TSC2 loss of heterozygosity in 7 of 13 (54%) of angiomyolipomas from sporadic LAM patients, suggesting that LAM and TSC could have a common genetic basis. In this study, we report the identification of somatic TSC2 mutations in five of seven angiomyolipomas from sporadic LAM patients. In all four patients from whom lung tissue was available, the same mutation found in the angiomyolipoma was present in the abnormal pulmonary smooth muscle cells. In no case was the mutation present in normal kidney, morphologically normal lung, or lymphoblastoid cells. Our data demonstrate that somatic mutations in the TSC2 gene occur in the angiomyolipomas and pulmonary LAM cells of women with sporadic LAM, strongly supporting a direct role of TSC2 in the pathogenesis of this disease. PMID- 10823954 TI - The complete unique long sequence and the overall genomic organization of the GA strain of Marek's disease virus. AB - We have determined the DNA sequence of the unique long (UL) region and the repeat long (RL) region in the genome of serotype 1 GA strain of Marek's disease virus (MDV), a member of the alpha-herpesvirus family. With this information, the complete nucleotide sequence of GA-MDV is now known. The entire GA-MDV genome is predicted to be about 174 kbp in size, with an organization of TRL-UL-IRL-IRS-US TRS, typical of a alpha-herpesvirus. The UL sequence contains 113,508 bp and has a base composition of 41.7% G + C. A total of 67 ORFs were identified completely within the UL region, among which 55 are homologous to genes encoded by herpes simplex virus-1. Twelve of them are unique with presently unknown functions. The sequence of RL reported here together with those published earlier reveal the major structural features of the RL. Virtually all of the ORFs encoded by RL are specific to serotype I of MDV. These ORFs are likely to contribute to some of the unique biological properties of MDV. Among the proteins encoded by MDV-specific ORFs are Meq, a jun/fos family of transcriptional factor implicated in transformation and latency, virus-encoded interleukin-8, a CXC chemokine, and pp38 and pp24, two phosphoproteins with undefined functions. There is also a putative lipase gene (LORF2) that has homologies in HPRS-24 (serotype II) strain of MDV and in various avian adenoviruses. An additional unique feature of MDV is the presence of long terminal repeat remnant sequences of avian retrovirus reticuloendotheliosis virus. These remnant sequences are derived from the U3 enhancer region through ancestral insertions by reticuloendotheliosis virus proviruses. PMID- 10823955 TI - Synaptophysin regulates clathrin-independent endocytosis of synaptic vesicles. AB - The GTPase dynamin I is required for synaptic vesicle (SV) endocytosis. Our observation that dynamin binds to the SV protein synaptophysin in a Ca(2+) dependent fashion suggested the possibility that a dynamin/synaptophysin complex functions in SV recycling. In this paper we show that disruption of the dynamin/synaptophysin interaction by peptide injection into the squid giant synapse preterminal results in a decrease in transmitter release during high frequency stimulation, indicating an inhibition of SV recycling. Electron microscopy of these synapses reveals a depletion of SVs, demonstrating a block of vesicle retrieval after fusion. In addition, we observed an increase in clathrin coated vesicles, indicating that the peptide does not block clathrin-dependent endocytosis. We conclude that the dynamin/synaptophysin complex functions in a clathrin-independent mechanism of SV endocytosis that is required for efficient synaptic transmission. PMID- 10823956 TI - Embryonic stem cells differentiate into oligodendrocytes and myelinate in culture and after spinal cord transplantation. AB - Demyelination contributes to the loss of function consequent to central nervous system (CNS) injury. Enhanced remyelination through transplantation of myelin producing cells may offer a pragmatic approach to restoring meaningful neurological function. An unlimited source of cells suitable for such transplantation therapy can be derived from embryonic stem (ES) cells, which are both pluripotent and genetically flexible. In this paper we show that oligodendrocyte cultures can be reliably produced from retinoic acid-induced ES cells and that these oligodendrocytes can myelinate axons in vitro. Methods were further developed for generating highly enriched cultures of oligodendrocytes through an additional culturing step, producing an intermediate "oligosphere" stage. To test whether ES cells can survive, migrate, and differentiate into mature myelin-producing cells in areas of demyelination in the adult CNS, ES cells were transplanted into the dorsal columns of adult rat spinal cord 3 days after chemical demyelination. In the demyelination site, large numbers of ES cells survived and differentiated primarily into mature oligodendrocytes that were capable of myelinating axons. Furthermore, when oligosphere cells were transplanted into the spinal cords of myelin-deficient shiverer (shi/shi) mutant mice, the ES cell-derived oligodendrocytes migrated into the host tissue, produced myelin and myelinated host axons. These studies demonstrate the ability of ES cell-derived oligodendrocytes to myelinate axons in culture and to replace lost myelin in the injured adult CNS. Transplantation of ES cells may be a practical approach to treatment of primary and secondary demyelinating diseases in the adult CNS. PMID- 10823957 TI - Classification of fusiform neocortical interneurons based on unsupervised clustering. AB - A classification of fusiform neocortical interneurons (n = 60) was performed with an unsupervised cluster analysis based on the comparison of multiple electrophysiological and molecular parameters studied by patch-clamp and single cell multiplex reverse transcription-PCR in rat neocortical acute slices. The multiplex reverse transcription-PCR protocol was designed to detect simultaneously the expression of GAD65, GAD67, calbindin, parvalbumin, calretinin, neuropeptide Y, vasoactive intestinal peptide (VIP), somatostatin (SS), cholecystokinin, alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid, kainate, N-methyl-d-aspartate, and metabotropic glutamate receptor subtypes. Three groups of fusiform interneurons with distinctive features were disclosed by the cluster analysis. The first type of fusiform neuron (n = 12), termed regular spiking nonpyramidal (RSNP)-SS cluster, was characterized by a firing pattern of RSNP cells and by a high occurrence of SS. The second type of fusiform neuron (n = 32), termed RSNP-VIP cluster, predominantly expressed VIP and also showed firing properties of RSNP neurons with accommodation profiles different from those of RSNP-SS cells. Finally, the last type of fusiform neuron (n = 16) contained a majority of irregular spiking-VIPergic neurons. In addition, the analysis of glutamate receptors revealed cell-type-specific expression profiles. This study shows that combinations of multiple independent criteria define distinct neocortical populations of interneurons potentially involved in specific functions. PMID- 10823958 TI - Direct activation of capsaicin receptors by products of lipoxygenases: endogenous capsaicin-like substances. AB - Capsaicin, a pungent ingredient of hot peppers, causes excitation of small sensory neurons, and thereby produces severe pain. A nonselective cation channel activated by capsaicin has been identified in sensory neurons and a cDNA encoding the channel has been cloned recently. However, an endogenous activator of the receptor has not yet been found. In this study, we show that several products of lipoxygenases directly activate the capsaicin-activated channel in isolated membrane patches of sensory neurons. Among them, 12- and 15-(S) hydroperoxyeicosatetraenoic acids, 5- and 15-(S)-hydroxyeicosatetraenoic acids, and leukotriene B(4) possessed the highest potency. The eicosanoids also activated the cloned capsaicin receptor (VR1) expressed in HEK cells. Prostaglandins and unsaturated fatty acids failed to activate the channel. These results suggest a novel signaling mechanism underlying the pain sensory transduction. PMID- 10823959 TI - Opposing effects of protein kinase C and protein kinase A on metabotropic glutamate receptor signaling: selective desensitization of the inositol trisphosphate/Ca2+ pathway by phosphorylation of the receptor-G protein-coupling domain. AB - Signaling by the metabotropic glutamate receptor 1alpha (mGluR1alpha) can lead to the accumulation of inositol 1,4, 5-trisphosphate (InsP(3)) and cAMP and to the modulation of K(+) and Ca(2+) channel opening. At present, very little is known about how these different actions are integrated and eventually turned off. Unraveling the molecular mechanisms underlying these functions is crucial for understanding mGluR-mediated regulation of synaptic transmission. It has been shown that receptor-induced activation of the InsP(3) pathway is subject to feedback inhibition mediated by protein kinase C (PKC). In this study, we provide evidence for a differential regulation by PKC and protein kinase A of two distinct mGluR1alpha-dependent signaling pathways. PKC activation selectively inhibits agonist-dependent stimulation of the InsP(3) pathway but does not affect receptor signaling via cAMP. In contrast, protein kinase A potentiates agonist independent signaling of the receptor via InsP(3). Furthermore, we demonstrate that the selectivity of PKC action on receptor signaling rests on phosphorylation of a threonine residue located in the G protein-interacting domain of the receptor. Modification at Thr(695) selectively disrupts mGluR1alpha-G(q/11) interaction without affecting signaling through G(s). Together, these data provide insight on the mechanisms by which selective down-regulation of a specific receptor-dependent signaling pathway can be achieved and on how cross talk between different second messenger cascades may contribute to fine-tune short- and long-term receptor activity. PMID- 10823960 TI - Cocaine vaccines: antibody protection against relapse in a rat model. AB - The efficacy of active immunization with the cocaine immunogen GNC-keyhole limpet hemocyanin (KLH) in preventing cocaine self-administration reinstatement was assessed in rats. An animal model of relapse was used where rats were trained to self-administer cocaine, subjected to a period of extinction by substituting the drug for saline, vaccinated, and re-exposed to cocaine. Compared with controls, animals immunized with GNC-KLH did not reinstate cocaine self-administration behavior when given a noncontingent cocaine infusion on two consecutive days. Upon double and triple infusions, 38-62% of vaccinated animals failed to reinstate as compared with full reinstatement in all control animals. Exposure to ad libitum cocaine reinstated baseline values in control animals and resulted in double to triple the baseline values of self-infusions in vaccinated animals, suggesting a partial antibody-mediated blockade of cocaine access to the central nervous system. This compensating effect was blocked by passive immunization pretreatment with the monoclonal IgG GNC92H2 in both vaccinated and control groups. To further assess the surmountability potential of GNC-KLH-induced antibody titers by cocaine self-administration, and the capacity of these titers to block the reinforcing effects of the drug, rats were tested at various doses of cocaine (0.015-0.5 mg/infusion). Active immunization with GNC-KLH produced approximately an 8-fold rightward shift of the dose-effect function for cocaine. The results reported suggest that immunopharmacotherapy may offer a promising means to treat cocaine abuse by aiding in the prevention of relapse. PMID- 10823961 TI - Thyroid hormone receptor-binding protein, an LXXLL motif-containing protein, functions as a general coactivator. AB - Nuclear hormone receptors activate gene transcription through ligand-dependent association with coactivators. Specific LXXLL sequence motifs present in these cofactors are sufficient to mediate these ligand-induced interactions. A thyroid hormone receptor (TR)-binding protein (TRBP) was cloned by a Sos-Ras yeast two hybrid system using TRbeta1-ligand binding domain as bait. TRBP contains 2063 amino acid residues, associates with TR through a LXXLL motif, and is ubiquitously expressed in a variety of tissues and cells. TRBP strongly transactivates through TRbeta1 and estrogen receptor in a dose-related and ligand dependent manner, and also exhibits coactivation through AP-1, CRE, and NFkappaB response elements, similar to the general coactivator CBP/p300. The C terminus of TRBP binds to CBP/p300 and DRIP130, a component of the DRIP/TRAP/ARC complex, which suggests that TRBP may activate transcription by means of such interactions. Further, the association of TRBP with the DNA-dependent protein kinase (DNA-PK) complex and DNA-independent phosphorylation of TRBP C terminus by DNA-PK point to a potential connection between transcriptional control and chromatin architecture regulation. PMID- 10823963 TI - Standing on large shoulders to advocate for children. PMID- 10823962 TI - A calcium-dependent protein kinase can inhibit a calmodulin-stimulated Ca2+ pump (ACA2) located in the endoplasmic reticulum of Arabidopsis. AB - The magnitude and duration of a cytosolic Ca(2+) release can potentially be altered by changing the rate of Ca(2+) efflux. In plant cells, Ca(2+) efflux from the cytoplasm is mediated by H(+)/Ca(2+)-antiporters and two types of Ca(2+) ATPases. ACA2 was recently identified as a calmodulin-regulated Ca(2+)-pump located in the endoplasmic reticulum. Here, we show that phosphorylation of its N terminal regulatory domain by a Ca(2+)-dependent protein kinase (CDPK isoform CPK1), inhibits both basal activity ( approximately 10%) and calmodulin stimulation ( approximately 75%), as shown by Ca(2+)-transport assays with recombinant enzyme expressed in yeast. A CDPK phosphorylation site was mapped to Ser(45) near a calmodulin binding site, using a fusion protein containing the N terminal domain as an in vitro substrate for a recombinant CPK1. In a full-length enzyme, an Ala substitution for Ser(45) (S45/A) completely blocked the observed CDPK inhibition of both basal and calmodulin-stimulated activities. An Asp substitution (S45/D) mimicked phosphoinhibition, indicating that a negative charge at this position is sufficient to account for phosphoinhibition. Interestingly, prior binding of calmodulin blocked phosphorylation. This suggests that, once ACA2 binds calmodulin, its activation state becomes resistant to phosphoinhibition. These results support the hypothesis that ACA2 activity is regulated as the balance between the initial kinetics of calmodulin stimulation and CDPK inhibition, providing an example in plants for a potential point of crosstalk between two different Ca(2+)-signaling pathways. PMID- 10823965 TI - Position statement on prevention of tobacco use in the pediatric population. PMID- 10823964 TI - Position statement on health risks and needs of gay, lesbian, bisexual, and transgender adolescents. PMID- 10823966 TI - Spring--enjoy it! PMID- 10823967 TI - Evaluation of sexual abuse in the pediatric patient. AB - Evaluating a patient for suspected child sexual abuse can be daunting for many pediatric primary care practitioners. The consequences of misdiagnosis can be devastating. Knowledge of common clinical presentations, both physical signs and symptoms and behavioral changes, is paramount. Sexual abuse allegations must be reported and investigated by child protection agencies or law enforcement. Practitioners must be aware of when and how to report suspected child sexual abuse, in addition to having a basic understanding of the medical examination and findings. With a caring, knowledgeable, and sensitive approach to allegations of sexual abuse, the practitioner can assist the child and his or her family through this very difficult process. PMID- 10823968 TI - Teen Club: a nursing intervention for reducing risk-taking behavior and improving well-being in female African American adolescents. AB - This article describes a nursing intervention called Teen Club that was designed to reduce risk-taking behavior and improve well-being in female African American adolescents. Participants were referred to Teen Club by their nurse practitioners, physicians, and a community health nurse who were working at an urban neighborhood health center's teen clinic. Referrals were based on factors such as parental substance abuse, lack of social and family support, and other characteristics thought to increase vulnerability to risk-taking behavior. The 2 year intervention included weekly group meetings co-led by a European American female community health nurse and a Latino American male community worker, supplemented by case management and home visits by both these persons. Findings from a retrospective group interview conducted with 11 of the 12 original participants are presented. This is the first step in a series of pilot studies designed to refine the Teen Club intervention in anticipation of a future prospective, randomized investigation of this health promotion and disease prevention model of nursing care. PMID- 10823969 TI - Risk and protective factors in children adopted from the former Soviet Union. AB - INTRODUCTION: The former Soviet Union (including the present independent republics of Russia, Ukraine, Latvia, Belarus, Lithuania, and Georgia) is the leading source of children adopted from overseas by persons in the United States (US Department of State, 1998). This study sought to (a) characterize the current social, academic, and conduct competencies of 6- to 9-year-old children adopted from the former Soviet Union who have resided in the United States for at least 2 years and (b) evaluate both risks and protective influences of adoptive families and their relationships to competence via a structural equation model. METHOD: Telephone interviews and a postal survey of children were drawn from a US community sample of 105 children. Measures included (a) Child Behavior Checklist, (b) Teacher Report Form, (c) Vineland Adaptive Behavior Scales, (c) Family Environment Scale, and (d) demographic information. RESULTS: Many children had experienced abuse, abandonment, or neglect between birth and entry to the institution. Their mean birth weight was 2637 g, and alcohol abuse by the birth mother was common (41%). Although the children scored below average in competence, adoptive family environments were positive and served as buffers between the risks experienced by the children and the subsequent development of competence within the adoptive family. CONCLUSION: Children's abilities ranged from severely challenged to developmentally normal. The high rate of fetal alcohol exposure in the children may portend future challenges for families. PMID- 10823970 TI - Factors influencing the immunization status of children in a rural setting. AB - INTRODUCTION: The purposes of this study were to (a) assess parental perceptions of their decision making regarding children's vaccinations and (b) describe parents' evaluation of immunization services provided by rural clinics/offices. METHODS: A qualitative design was used in this study, which was conducted in rural Missouri. Twelve mothers of children younger than age 3 years with fewer than the recommended number of immunizations were interviewed using a semi structured format. The interview results were analyzed using the constant comparative method. RESULTS: The following parental perceptions were identified as factors related to immunizations in this rural setting: knowledge of communicable diseases and vaccines, misperceptions about communicable diseases and vaccines, past experiences, competing tasks, transportation, health care personnel, need for reminders, health system, and cost. DISCUSSION: Two findings unique to this study were the importance of relationships with health care providers and the challenge of competing tasks. These findings, combined with the other factors identified, reinforced the importance of rural health care providers' maintaining a strong relationship with clients, providing accurate and timely information, and ensuring a readily accessible health care system. PMID- 10823971 TI - What parents of children with asthma tell us. AB - INTRODUCTION: The family's contribution to effective asthma management is increasingly being recognized. This study gathered and analyzed information from parents about their greatest fears relative to having a child with asthma and identified information that parents believed was critical for providers to acknowledge. The methodology serves as a model for possible practitioner-directed action research with their care population. METHODS: Written information from 52 parents who attended focus groups comprise the data. Data were analyzed using constant comparative strategies. RESULTS: Common parental fears included uncertainty, long-term effects of medication, and concern that the condition would not go away or improve. Common themes parents wanted providers to know included the following: parents need more information; parents are scared and fearful; living with asthma is difficult, and this stress affects the parents' behavior; and parents know what is best for their child. Age trends were identified for fear themes only. DISCUSSION: Findings suggest parents can and should be asked about their greatest fears and concerns. Although developmental patterns may be used to initiate discussion, providers need to identify parents' unique concerns so they can integrate them in refining the child's asthma action plan. PMID- 10823972 TI - Cardiac arrhythmias in children. PMID- 10823973 TI - Screening and treatment of sexually transmitted diseases Part 2: trichomonas, human papillomavirus infection, and genital herpes simplex virus. PMID- 10823974 TI - An adolescent with osteoporosis. PMID- 10823975 TI - Improving advocacy through new technology. PMID- 10823976 TI - Study compares performance of nurse practitioners and physicians. PMID- 10823978 TI - Questions & answers. PMID- 10823977 TI - Nurses who lead the way. PMID- 10823979 TI - Malnutrition in elderly people newly admitted to a community resident home. AB - The purpose of this study was to assess nutritional status in elderly people, newly admitted to a community resident home, and to describe the characteristics of residents with protein-energy malnutrition (PEM). The participants were 261 people, 65 to 103 years of age (M= 84.8+/-7.1 in women, 82.5+/-6.4 in men) who during one year entered special types of housing for the elderly in a municipality in the south of Sweden. During the first two weeks after admission nutritional status was assessed using weight index, triceps skinfold thickness, arm muscle circumference, serum albumin and transthyretin. Demographic and sociomedical data and eating-related factors were collected by using structured interviews and studying residents records. PEM was found in 29% of the residents who entered municipal care from their own homes, 33% among those moving within municipal care and 43% of the residents who entered from hospital care. Pressure sores or leg ulcers, psychological stress or acute disease in the previous 3 months, reduced fluid intake, deteriorated appetite, reduced mobility, need of help during meals and gastrointestinal symptoms were factors associated with PEM. PMID- 10823980 TI - Correlates of bone mineral density in nursing home residents. AB - PURPOSE: To explore correlates of bone mineral density (BMD) in nursing home residents, examining factors associated with BMD in community cohorts and factors prevalent among nursing home residents. METHODS: A cross sectional study of 2156 elderly residents from a representative state-wide sample of 47 nursing homes in Maryland. Data consisted of BMD scans of the distal radius and ulna; information on comorbidities, activities, behavior and medications abstracted from medical charts and interviews; and observation of cognitive and neuromuscular performance. RESULTS: Forty-two percent of the variance in BMD was explained by age, gender, race and weight. Fracture, use of steroids and seizure medications, and alcohol use were also related to low BMD. Other factors prevalent in nursing home populations, such as cognitive impairment and depressive symptomatology, were not strongly associated with low BMD. CONCLUSIONS: Despite the diversity of the nursing home population, their older age and greater dependency and comorbidity, the factors most closely related to low BMD in nursing home residents were comparable to those in community-dwellers. Demographic information, along with other clinical factors such as fracture and use of steroid and seizure medications, may aid health care providers in identifying high risk individuals in nursing homes. PMID- 10823981 TI - Epidemiological study of malnutrition in elderly patients in acute, sub-acute and long-term care using the MNA. AB - MNA is a simple and accurate way to assess the nutritional status in routine practice, and is suitable for systematic use and large epidemiologic studies. The purpose of this study was to evaluate the difference in the nutritional status of elderly patients hospitalized in different types of care in the same hospital, and to evaluate its relationship with risk factors. Nutritional status was evaluated in 918 elderly patients hospitalized in acute care (AC) (n=299), sub acute care (SAC) (n=196) or long-term care (LTC) (n=423), using the MNA (Mini Nutritional Assessment), a nutritional assessment tool including 18 items grouped in 4 domains, within the first 48 h after admission (all subjects) and at the end of hospitalization (AC, SAC). More patients were rated in the "malnourished" class in SAC (32.5%) than in AC (24.5%) and LTC (24. 7%). Retrospective analysis showed that the initial nutritional status was linked to the type of care and the nature of underlying pathology. The nutritional status on arrival was worse in patients in SAC, and better in those in LTC (p = 0.005). This is probably due to a difference in the kind of patients hospitalized. The nutritional status was worse in patients suffering from infectious disease, stroke, dementia and traumatic injuries, and, conversely, better in patients suffering from cardiopathy, metabolic and gastro-intestinal (except cancerous) diseases (p < 0.0001). Prospective analysis showed that duration of hospitalization was the only variable found to be linked to an improvement of nutritional status. The MNA is a rapid, effective and cheap tool for the assessment of nutritional status and moreover for evaluation of the mortality risk of patients admitted into AC and SAC. PMID- 10823982 TI - Knowledge and use of the food label among senior women in the management of type 2 diabetes mellitus. AB - Food selection is a key component in the management of diabetes. The foods selected affect the caloric and nutrient composition of the diet and blood glucose levels among people with diabetes. The nutrition information on the food label can guide decisions for food purchases. Whether senior women with diabetes use and comprehend the nutrition information on the food label has not been determined. Therefore, the purpose of this study was to assess knowledge and beliefs about the food label and diabetes management among women with type 2 diabetes mellitus =65 years of age. Focus groups were conducted and participants (n=24) completed a knowledge test about the food label. Analysis involved extensive review of the videotapes and transcripts of the focus groups. Participants reported that they referred to the nutrition information on the label when grocery shopping. Yet, comprehension of terms and product claims on the label was poor. The mean score on the food label knowledge test was only 49%. Misconceptions about the nutritional management of diabetes also were noted. Thus, senior women with diabetes need additional education about the food label to facilitate application of the information in meal planning and to manage diabetes successfully. PMID- 10823983 TI - Effect of taste and smell on secretion rate of salivary IgA in elderly and young persons. AB - Two experiments were performed to determine the effect of taste and odor stimuli on secretion rate of salivary IgA in young and elderly individuals. In Experiment 1, three stimuli were applied to the tongue: 1) "flavor" drops (60% sugar, 5% cocoa powder, and 0.1 % Irish cream odor); 2) sugar (60%); and 3) water. In Experiment 2, four different foods (chicken broth, onion soup, corn, and carrots) were tested with and without monosodium glutamate (MSG). The stimuli in both experiments were delivered three times in a one hour period: t=0, t=30 minutes, t=60 minutes. The results of Experiment 1 indicated that application of sugar (taste alone) and flavor (taste and odor combined) to the tongue induced significantly higher secretion rates of sIgA than the application of water in both elderly and young subjects. In addition, flavor application produced significantly higher secretion rates of sIgA than sugar application alone. Secretion rates of sIgA in young persons were significantly higher than those in elderly persons. In Experiment 2, the increase in secretion rate of sIgA at 30 and 60 minutes for each food with MSG was greater than the same food without MSG for the elderly subjects. The increases in secretion rates of sIgA were produced by elevated salivary flow (Experiments 1 and 2) as well as increased absolute concentrations of sIgA (Experiment 1). The elevation of absolute concentrations of sIgA by chemosensory stimuli may involve neural-immune connections. The improvements in salivary flow and secretory immunity by repeated taste and smell stimulation found here have clinical potential for treatment of immune deficiencies and dry mouth which frequently occur in elderly individuals. PMID- 10823984 TI - Antibodies against the synthetic reactive site (SER338-ILE353) of human PAI-1 neutralize PAI-1 activity. PMID- 10823985 TI - Whose rights supersede, those of the patient or those of the unborn fetus? PMID- 10823987 TI - Surfing the healthcare Web. PMID- 10823986 TI - Case study: the interaction of patient and nurse suffering. PMID- 10823989 TI - Hospital executives and ethics committees: an effective collaboration. AB - Should hospital and healthcare executives participate on their organization's ethics committee? This question becomes more relevant in the current healthcare environment as nurses and physicians assume more managerial responsibilities. This article reviews functions of the ethics committee and discusses moral, conceptual, and practical issues surrounding managerial participation on these committees. The authors conclude that executive management's participation on ethics committees is both appropriate and necessary in the current healthcare environment. PMID- 10823988 TI - Home healthcare under fire: fraud and abuse. AB - Fraudulent and abusive practices in home healthcare are costly. They damage the integrity of the home health agency, the home care industry, and home care providers. Federal and state enforcement activities have increased exponentially over the past few years to fight fraud and abuse, resulting in the collection of nearly $1.1 billion in fines, settlements, and administrative impositions in 1997. This article discusses the issues of fraud and abuse and the responsibilities of home healthcare providers toward prevention and control of such practices. PMID- 10823990 TI - Above all, do no harm. PMID- 10823991 TI - A call to arms. PMID- 10823992 TI - Healthcare on-line. Agency for Healthcare Policy and Research. PMID- 10823993 TI - The False Claims Act. An interview with Mark Kleiman. AB - JONA's Healthcare Law, Ethics, and Regulation interviewed Attorney Mark Allen Kleiman regarding the False Claims Act. Mr. Kleiman has had much experience with this Act in both prosecution and consulting. In this interview, he provides great insight into the details and interpretations of the Act, as well as implications for healthcare providers. PMID- 10823994 TI - Elderly patients' understanding of advance directives. AB - Growing evidence suggests that advance directives (ADs) are often ineffective. Further explanation of how these documents are viewed by the public, the expectations of how decisions about treatment are made, and, in particular, patients' understanding of how ADs are actually used in the clinical setting, are warranted. This article details a descriptive study in which patients were interviewed, during hospital stays, about their beliefs and understanding of advanced directives, as well as the processes used in completing them. The study was undertaken in a community hospital located in a rural area in the Midwest. Findings show that many patients were able to clearly articulate what an AD means in terms of making their choices known. However, misconceptions were found in patients' understanding of ADs and only 46% reported discussing their ADs with a physician. Strategies for effective implementation of education programs related to ADs are detailed here. PMID- 10823995 TI - Healthy people--a new millennium. Progress and comparison on the Healthy People 2000 and Healthy People 2010 objectives. AB - An understanding of public health and its impact on the health of the nation is vital to anyone involved in implementing change in healthcare. The Healthy People Initiatives are the basis for change in an extraordinarily large segment of the healthcare population. These visions of public health are not simply an idea of the perfect health services, but are a very detailed plan of desired outcomes and the means to reach them. Availability of healthcare services to all segments of our population is an ethical obligation to anyone with a true vocation in healthcare. This article presents a brief overview of the Healthy People Initiative. PMID- 10823996 TI - Touched by an angel of light: care at the end of life. PMID- 10823997 TI - Malpractice litigation trends for the new millennium. PMID- 10823998 TI - A student nurse's perception of right and wrong. PMID- 10823999 TI - Healthcare on-line. PMID- 10824000 TI - Spotlight on ... the American Organization of Nurse Executives. The new nursing shortage. AB - The American Organization of Nurse Executives (AONE) strives to provide its members with timely and useful information. In this article, the author describes AONE's work to guide the policy process about nurse shortages. The results of this survey will help you practice in your position and in your state, regional, and national policy work. AONE's policy agenda is driven by its mission and commitment to patient care. One of the most current, compelling policy issues is ensuring an adequate supply of nurses to meet the patient care demand. In response to the increasing reports of nurse shortages, AONE engaged the HSM Group, Ltd. to conduct a survey to determine if hospitals were experiencing a shortage. The survey questions were developed with the Health Resources and Services Administration (HRSA), Division of Nursing, the American Nurses Association, and the American Society for Healthcare Human Resources of the American Hospital Association. The Research on Nursing Staff Shortage was published in 1999. PMID- 10824001 TI - Nursing law violations. A threat to competent and safe nursing practice. AB - Nurse Practice Act violations pose threats to consumers of nursing services and lead to disciplinary actions against nurses by boards of nursing. To analyze nursing law violations, the actions and decisions of boards of nursing, and evaluate trends in negligent and unsafe nursing practice, the authors reviewed nursing law violations as well as rates of recidivism among nurses who received actions against their nursing licenses in Kentucky. The authors discuss how their findings can assist nurse administrators in investigating nurse care givers before employment and in initiating safeguards against nurse violations that affect client safety. PMID- 10824002 TI - Effective employee relations in reengineered organizations. AB - The degree to which a healthcare organization can effectively respond to changes that are brought about by managed care is often the result of how well the organization engages the workforce in the changes. The goal of a process-centered healthcare organization is to deliver service that is truly patient-centered. Successful organizations understand that the actualization of this goal begins with the decision to engage in an employee-guided approach to reengineering. This article describes issues of effective employee relations in the postreengineering era. Strategies for dealing effectively with labor unions or for preventing unionization are also discussed. PMID- 10824003 TI - A century of healthcare delivery. PMID- 10824004 TI - The small hospital dilemma. PMID- 10824005 TI - Healthcare on-line. PMID- 10824007 TI - Solving the healthcare crisis: a sociophilosophical approach. PMID- 10824006 TI - Who killed Georgette Smith? Healthcare providers at the intersection of criminal law and patient care. PMID- 10824008 TI - But is it really ART? Economic, legal, and ethical implications of infertility technology. PMID- 10824009 TI - The patient who could not be discharged. How far should patient autonomy extend? AB - A male patient was admitted to the acquired immune deficiency syndrome (AIDS) unit for hemodialysis. His history revealed that he was homeless and that he had tested positive for human immunodeficiency virus (HIV+). He also had a history of alcohol and intravenous drug abuse and tuberculosis. Based on the results of a chest X-ray, he was placed in respiratory isolation. During the next few days of his hospitalization, he exhibited nonadherent behavior toward the treatment regime. Because of previous verbal and physical abuse to staff and patients, all local hemodialysis centers refused to accept him as a patient. Thus, he became a patient who seemingly could never be discharged. A discussion related to the theoretical and practical scope of patient autonomy, institutional altruism vs. institutional self-interest, and the need for social policy to facilitate a just and humane resolution to this ethical situation is presented here. PMID- 10824010 TI - Year 2000: this moment in time! PMID- 10824011 TI - National Center for Complementary and Alternative Medicine. PMID- 10824012 TI - Mergers and acquisitions. Frequently asked questions and answers. AB - This article is structured in a question/answer format based on interviews with Dr. Carolyn Hope Smeltzer and Salima Manji Lin of PricewaterhouseCoopers, Chicago, and Chuck Thomas of Hinshaw & Culbertson, Rockford. The questions come from CEO's, healthcare executives, and nurse executives at hospitals that are contemplating mergers or that have both succeeded and failed to merge their institutions. The experts share their knowledge. PMID- 10824013 TI - State and territorial boards of nursing approaches to the use of unlicensed assistive personnel. AB - This study examined U.S. state and territorial boards of nursing approaches to the regulation of the use of: unlicensed assistive personnel (UAP) in acute care hospitals; state and jurisdictional authority, oversight and disciplinary action related to registered nurse (RN) delegation, supervision and assignment; educational preparation requirements for UAP; and future projections for their use. A survey was administered to 53 state and territorial boards of nursing officials in 1998. A majority of the states reported that they had regulations/guidelines for RN's who supervised UAP and regulations that protected the use of the RN title. Few states used the American Nurses Association or National Council of State Boards of Nursing definitions for delegation, supervision, or assignment. The majority have formulated their own definitions. The majority of states reported no standardized curriculum in place for UAP employed in acute care hospitals. More than half of the states reported that no plans existed for developing a curriculum. PMID- 10824014 TI - Is it ethical to practice intubations on the deceased? AB - One-third to one-half of emergency departments in the United States and Australia perform endotracheal intubations (ETI's) on the newly dead. Sixty-three percent of emergency medicine and 58% of neonatal critical care training programs allowed procedures to be performed on patients after death; only 10% of these programs required family consent for this practice. This article reviews the arguments for and against this ethical issue. A case study is included to highlight the issue's complexity, and to assist readers in identifying their beliefs (and those of their institutions) about the tissue. An overview of ethically related terms, definitions, and theories and a decision-making model are included to establish a knowledgeable baseline for dealing with any ethical issue. PMID- 10824015 TI - Ethical decision making in nurses. Relationships among moral reasoning, coping style, and ethics stress. AB - Stress related to ethical decision-making is a serious consequence of frequent encounters with ethical dilemmas for oncology nurses. A descriptive, correlational design using survey techniques was used as a study design with a nationwide sample of 229 oncology nurses. The results indicated nurses experienced an average of 32 different types of ethical dilemmas within the past year on a daily basis. Pain management is the most frequently cited ethical dilemma, followed by cost containment issues and making quality of life and other decisions in the patient's best interest. Approximately 80% of respondents rated their ethics stress level as a 6 or above on a scale of 0 to 10. Forty-three percent of the sample indicated they use an independent or "sovereign" style of moral reasoning, 23% rely on or accommodate to the judgment of others, and 34% use characteristics of both moral reasoning styles. Understanding the relationships among style of moral reasoning, coping style, and ethics stress can assist nurses and administrators to deal more effectively with the increased moral distress found in many oncology practice settings today. Findings suggest specific interventions for reducing ethics stress in this population of nurses. PMID- 10824016 TI - Accurately analyzing aqueous flare data. PMID- 10824017 TI - Heterogeneity in Individual Mortality Risk and Its Importance for Evolutionary Studies of Senescence. AB - Mortality was simulated under the assumption of heterogeneity in individual age specific mortality risk. Heterogeneity was modeled by assigning each individual its own Gompertz mortality function. Means and variances of the Gompertz intercept and slope parameters were based on published data for Drosophila melanogaster. Simulations of large cohorts reproduced mortality plateaus similar to those observed for actual cohorts of flies. Catastrophic late-age mortality was not observed except when heterogeneity was very low and rates of senescence were very high. A second set of simulations was designed to mimic experiments that have investigated age-specific patterns of genetic variance in mortality rates. Within-genotype heterogeneity in mortality risk resulted in a decline in genetic variance of mortality rates at old ages. That result suggests that the decline in genetic variance at old ages that has been observed in some experiments is an artifact of heterogeneity. Mortality rate plateaus, decrease in genetic variance of mortality rates at old ages, and absence of catastrophic late age mortality all appear to contradict predictions of the evolutionary theory of senescence. Heterogeneity in mortality risk may explain those contradictions. PMID- 10824018 TI - Strategy Space and the Disturbance Spectrum: A Life-History Model for Tree Species Coexistence. AB - The disturbance spectrum consists of disturbance patterns differing in type, size, intensity, and frequency. It is proposed that tree life-history traits are adaptations to particular disturbance regimes. Four independent axes are proposed to define the dominant dimensions of tree strategy space: shade tolerance, tree height, capacity for vegetative reproduction, and seed dispersal distance. A fitness model was developed to elucidate interactions between the proposed life history traits. The model shows how alternate life-history sets can coexist when disturbance patterns fluctuate in space and time. Variable disturbance regimes were shown, based on data and simulation results, to enhance species coexistence, as predicted. The strategy space model accurately predicts the number of common tree species for the eastern United States, boreal Canada, and southwestern pinon juniper woodlands. The model also provides an explanation for latitudinal gradients in tree species richness in North America and Europe. The proposed model predicts a relationship between disturbance characteristics and the species composition of a forest that allows for the coexistence of large numbers of species. The life-history traits of size, growth rate, life span, shade tolerance, age of reproduction, seed dispersal distance, and vegetative reproduction are all incorporated into the model. PMID- 10824019 TI - The Effect of Assortative Mating on the Coexistence of a Hybridogenetic Waterfrog and Its Sexual Host. AB - In central Europe, the hybridogenetic waterfrog Rana esculenta, a hybrid between Rana ridibunda and Rana lessonae, lives in sympatry with one of its parental species, the poolfrog Rana lessonae. As R. esculenta has to backcross constantly with R. lessonae in order to produce viable offspring, this coexistence is obligatory for R. esculenta. Since R. esculenta has a higher primary fitness than R. lessonae, a mechanism is required that prevents the hybrid from driving the parental species, and hence itself, to extinction. Here, we present an analytical model and a computer simulation that investigate whether assortative mating can operate as a such a control mechanism. Our results show that assortative mating is very effective in regulating coexistence in such a hybrid-host system. This is particularly true when choice is affected by the proportion of the two male types in the population. Furthermore, we could show that even if the species composition in a mixed hybrid-host population may be largely influenced by differences in life-history parameters, assortative mating still plays a very important role by stabilizing coexistence. Thus, mating behavior turns out to be more important for the populations dynamics of hybridogenetic waterfrog systems than previously assumed. PMID- 10824020 TI - Seed Size, Fruit Size, and Dispersal Systems in Angiosperms from the Early Cretaceous to the Late Tertiary. AB - Fossil data from 25 angiosperm floras from the Early Cretaceous (~124 million years ago) to the Pliocene (~2 million years ago) were compiled to estimate sizes of seeds and fruits and the relative proportion of two different seed-dispersal systems by animals and by wind. The results suggest that, first, seed and fruit sizes were generally small during most of the Cretaceous, in agreement with previous suggestions, but the trend of increasing sizes started before the Cretaceous-Tertiary boundary; second, there was a decrease in both seed and fruit sizes during late Eocene and Oligocene, reaching a level that has continued to the Late Tertiary; third, the fraction of animal dispersal was, in contrast to previous suggestions, rather high also during the Cretaceous but increased drastically in the Early Tertiary and declined congruently with the declining seed and fruit sizes from the late Eocene; and fourth, the fraction of wind dispersal showed a bimodal pattern, being high in the Late Cretaceous and in the Oligocene-Miocene but with a drop in between. We find that the observed trends are only weakly related to the availability of animal fruit dispersers. Instead, the trends are congruent with a climate-driven change in environmental conditions for recruitment, where larger seeds are favored by closed forest vegetation. The prevalence of semiopen, dry, and probably herbivore-disturbed vegetation during the Cretaceous, the development of closed multistratal forests in the Eocene, and the later development of a more open vegetation and grasslands starting in the Oligocene-Miocene, are reflected in the distribution of angiosperm seed and fruit sizes and in the dispersal systems. PMID- 10824021 TI - Geographic Variation in Camouflage Specialization by a Decorator Crab. AB - In North Carolina, the decorator crab Libinia dubia camouflages almost exclusively with the chemically noxious alga Dictyota menstrualis. By placing this alga on its carapace, the crab behaviorally sequesters the defensive chemicals of the plant and gains protection from omnivorous consumers. However, Dictyota is absent north of North Carolina, whereas Libinia occurs as far north as New England. Crabs from three northern locations where Dictyota is absent (Rhode Island, Connecticut, and New Jersey) camouflaged to match their environment, rather than selectively accumulating any one species. When D. menstrualis was offered to crabs from northern sites, they did not distinguish between it and other seaweeds for camouflage, whereas crabs from Alabama and two locations in North Carolina used D. menstrualis almost exclusively. In addition, in winter and spring, when Dictyota was seasonally absent in North Carolina, Libinia selectively camouflaged with the sun sponge Hymeniacidon heliophila, which was chemically unpalatable to local fishes. Thus, southern crabs were consistent specialists on chemically defended species for camouflage, while northern crabs were more generalized. The geographic shift in crab behavior away from specialization coincides with a reported decrease in both total predation pressure and the frequency of omnivorous consumers. These shifts in the nature and intensity of predation pressure may favor different camouflage strategies (generalist vs. specialist), contributing to the observed geographic differences in camouflage behavior. PMID- 10824022 TI - High and Dry: Drought Stress, Sex-Allocation Trade-offs, and Selection on Flower Size in the Alpine Wildflower Polemonium viscosum (Polemoniaceae). AB - Sex-allocation trade-offs may maintain variation in secondary sexual characteristics if such traits vary in their benefits or costs in association with different genders. In Polemonium viscosum, large flowers benefit both male and female aspects of reproduction. In this study, I explore how resource investment in flower size influences the cost of allocation to male and female function. Large flowers exact a water cost in P. viscosum under dry conditions. In an extreme drought in 1997, experimentally watered plants had higher survival and fecundity than controls. By comparing allocation patterns between plants dying from drought and survivors, I tested whether the demographic cost of large flowers increases with allocation to fecundity. Controls that died showed a positive relationship between flower size and fruit production, while survivors showed a negative relationship or trade-off. Watered plants showed no such trade off. To test whether drought affects the relationship of corolla size to male function, I used leaf-water potential in 1998 to classify plants as stressed or unstressed. Corolla size showed positive correlations to pollen per flower regardless of drought stress. I conclude that under drought the demographic cost of producing large flowers is gender dependent, such that viability selection favors either small-flowered plants with female-biased reproduction or larger flowered plants with male-biased reproduction. PMID- 10824023 TI - Vertebrate Secondary Sexual Characteristics-Physiological Mechanisms and Evolutionary Patterns. AB - As most commonly presented, the organization-activation theory of sexual differentiation emphasizes the importance of the relative age of the organism for understanding steroid hormone effects. However, considering the actual physiological mechanisms of tissue enlargement (hyperplasia/hypertrophy) provides an additional perspective for interpreting patterns of evolutionary change in sexual dimorphism. Using that focus, it is possible to suggest mechanistic explanations for patterns of allometry and the size of some secondary sexual characteristics produced by "runaway" selection. It can also lead to the formulation of testable hypotheses regarding the type of sexually dimorphic characters that might evolve through "good genes" models; the relationship between intrasexual competition, polygyny, and the development and size of male weapons; and the control and type of secondary sexual characteristics that will be present in males with associated and disassociated reproductive patterns. PMID- 10824024 TI - Alkaloid Uptake Increases Fitness in a Hemiparasitic Plant via Reduced Herbivory and Increased Pollination. AB - It has been historically difficult to manipulate secondary compounds in living plants to assess how these compounds influence plant-herbivore and plant pollinator interactions. Using a hemiparasitic plant that takes up secondary compounds from host plants, I experimentally manipulated secondary compounds in planta and assessed their effects on herbivores and pollinators in the field. Here, I show that the uptake of alkaloids in the annual hemiparasite Castilleja indivisa resulted in decreased herbivory, increased visitation by pollinators, and increased lifetime seed production. These results indicate that resistance traits such as alkaloids can increase plant fitness directly by reducing herbivore attack and indirectly by increasing pollinator visitation to defended plants. Thus, selection for production of secondary compounds may be underestimated by considering only the direct effect of herbivores on plant fitness. PMID- 10824025 TI - Forced Matings in Natural Populations of Drosophila. PMID- 10824026 TI - Comparative in vitro activity of gemifloxacin. AB - Gemifloxacin is a new quinolone and, like moxifloxacin, trovafloxacin, grepafloxacin and clinafloxacin, is more potent in vitro than ciprofloxacin or ofloxacin against Gram-positive aerobes. Gemifloxacin was the most potent of the quinolones tested against streptococci and most ciprofloxacin-resistant pneumococci were susceptible to gemifloxacin. Gemifloxacin, like moxifloxacin, trovafloxacin, grepafloxacin and clinafloxacin, was more potent than ciprofloxacin or ofloxacin against all staphylococci and many ciprofloxacin resistant isolates were susceptible to these quinolones. Against Gram-negative aerobes gemifloxacin was as potent as or slightly less potent than ciprofloxacin, and isolates resistant to ciprofloxacin were also resistant to gemifloxacin and to moxifloxacin, trovafloxacin and grepafloxacin. Gemifloxacin was also the most potent quinolone against Gram-positive anaerobes and fusobacteria but trovafloxacin was the most potent agent tested against other Gram-negative anaerobes. PMID- 10824027 TI - In vitro activity of gemifloxacin against a broad range of recent clinical isolates from the USA. AB - The antibacterial potencies of gemifloxacin (SB-265805) and 13 comparator compounds were determined by broth microdilution against a panel of 645 Gram positive and 995 Gram-negative organisms collected from various USA sites. Time kill studies were performed and postantibiotic effect (PAE) was determined for several organisms using trovafloxacin and ciprofloxacin as comparator compounds. Based on MIC(90)s, gemifloxacin was the most potent compound tested against Gram positive isolates: Streptococcus pneumoniae (MIC(90) 0. 016 mg/L), Streptococcus agalactiae (0.03 mg/L), Streptococcus pyogenes (0.03 mg/L), viridans streptococci (0.12 mg/L), methicillin-susceptible Staphylococcus aureus (0.03 mg/L), Staphylococcus epidermidis (2 mg/L), Staphylococcus saprophyticus (0. 016 mg/L) and Enterococcus faecalis (2 mg/L). Against Gram-negative isolates, the potency of gemifloxacin was equal to that of levofloxacin and ciprofloxacin and generally better than that of ofloxacin, grepafloxacin, trovafloxacin and nalidixic acid. MIC(90)s for gemifloxacin were: Haemophilus influenzae (6 h at 4 x MIC) and shorter PAEs with E. faecalis (0.1-0.6 h) and K. pneumoniae (0.1-0.2 h). In conclusion, gemifloxacin is a novel quinolone with a broad spectrum of antimicrobial activity. It has substantially improved potency against Gram-positive organisms, especially streptococci, for which gemifloxacin is generally at least eight- to 16-fold more potent than other quinolones tested. It retains the good Gram-negative activity seen with ciprofloxacin and levofloxacin and shows good bactericidal activity and prolonged PAEs. PMID- 10824028 TI - In vitro antibacterial activity of gemifloxacin and comparator compounds against common respiratory pathogens. AB - This study investigated the in vitro potency of the novel quinolone agent gemifloxacin (SB-265805), in comparison with other quionolones, beta-lactams, macrolides and trimethoprim- sulphamethoxazole, against a panel of common respiratory pathogens. This panel comprised recent clinical isolates of Streptococcus pneumoniae (n = 347), Haemophilus influenzae (n = 256) and Moraxella catarrhalis (n = 184). Overall, the quinolones were highly active against H. influenzae and were the most potent agents against M. catarrhalis. Gemifloxacin was the most potent quinolone tested against all three species and was four- to 512-fold more potent against pneumococci than trovafloxacin, grepafloxacin, levofloxacin, ciprofloxacin, ofloxacin, gentamicin, cefuroxime, penicillin, ampicillin, clarithromycin, azithromycin or trimethoprim- sulphamethoxazole. Against 19 ofloxacin-intermediate and 52 ofloxacin-resistant strains of S. pneumoniae, gemifloxacin retained activity, and was the only agent tested with MICs of < or =0.5 mg/L. The results of this study demonstrate the excellent in vitro antibacterial activity of gemifloxacin against pathogens commonly associated with respiratory tract infections and suggest that gemifloxacin has significant potential in the treatment of such infections, including those caused by pneumococci considered resistant to other quinolones. PMID- 10824029 TI - Comparative potency of gemifloxacin, new quinolones, macrolides, tetracycline and clindamycin against Mycoplasma spp. AB - This study determined the comparative in vitro potency of the new investigative quinolone gemifloxacin (SB-265805) using low-passaged clinical isolates and type strains of mycoplasma commonly found in the human respiratory and urogenital tracts. Organisms studied were Mycoplasma pneumoniae, Mycoplasma hominis, Mycoplasma fermentans, Mycoplasma genitalium, Mycoplasma penetrans and Ureaplasma urealyticum, obtained from different geographical regions. Comparator drugs were levofloxacin, trovafloxacin, grepafloxacin, azithromycin, clarithromycin, tetracycline and clindamycin. MICs were determined using a microbroth dilution method. The overall range of MICs of gemifloxacin was < or =0.008-0.125 mg/L for different Mycoplasma spp. and < or =0.008-0.5 mg/L for Ureaplasma spp. Depending on the species tested, gemifloxacin showed variable results when compared with the macrolides. However, gemifloxacin was as potent as, or more potent than, tetracycline, clindamycin and the other quinolones investigated. PMID- 10824030 TI - Activity of gemifloxacin and other new quinolones against Chlamydia pneumoniae: a review. AB - Quinolones are currently used as empirical therapy for treatment of community acquired lower respiratory infections as they are effective against a broad range of conventional bacterial and 'atypical' pathogens, including Chlamydia pneumoniae. C. pneumoniae is estimated to be associated with 10-20% of community acquired pneumonia in adults, and has recently been suggested to play a role in several non-respiratory conditions, including atherosclerosis. The newer, third generation quinolones have enhanced activity against Gram-positive bacteria, including Streptococcus pneumoniae, and prolonged serum half-lives that permit once-daily dosing. Although gemifloxacin (SB-265805) and other new quinolones have good activity against C. pneumoniae in vitro, practically all published treatment studies have relied on serological diagnosis. Consequently, the microbiological efficacy of these agents in human infection has not been assessed. This paper reviews what is known to date of the in vivo microbiological efficacy of the quinolones against C. pneumoniae, and demonstrates the importance of assessing this parameter when evaluating the clinical utility of these agents in C. pneumoniae infection. PMID- 10824031 TI - Comparative in vitro activity and post-antibiotic effect of gemifloxacin against Legionella spp. AB - The comparative in vitro potency and post-antibiotic effect (PAE) of gemifloxacin (SB-265805), moxifloxacin, trovafloxacin, grepafloxacin, levofloxacin, ofloxacin, ciprofloxacin, azithromycin, clarithromycin, erythromycin and rifampicin were evaluated against Legionella pneumophila serogroups 1-9 and 12 (n = 204) and other Legionella spp. (n = 34). MICs were determined by standard two-fold agar dilution. PAE was determined by exposing the isolates to the test agents at 4 x MIC for 1 h. Trovafloxacin was the most potent agent overall (MIC range < or =0.004-0.016 mg/L, MIC(90) < or =0.008 mg/L). Of the other quinolones tested, gemifloxacin, moxifloxacin, grepafloxacin and levofloxacin were more potent (MIC(90) 0.016 mg/L) against L. pneumophila than ciprofloxacin and ofloxacin (MIC(90) 0. 03 mg/L). Against Legionella spp., the test quinolones were more potent (MIC range < or =0.004-0.06 mg/L) than either erythromycin or azithromycin (MIC(90) 0.5 and 0.25 mg/L, respectively). Gemifloxacin had the longest PAE (4.65 h) of the agents tested against erythromycin-resistant L. pneumophila. Of the quinolones, only gemifloxacin, grepafloxacin, levofloxacin and ofloxacin had PAEs of >3 h against erythromycin-resistant Legionella spp. Azithromycin, erythromycin and clarithromycin had PAEs of <3 h against all erythromycin-resistant strains. Against erythromycin-susceptible L. pneumophila, only gemifloxacin, moxifloxacin, ofloxacin and ciprofloxacin had PAEs of >3 h, and only rifampicin, ofloxacin, gemifloxacin and erythromycin had PAEs of >2 h against erythromycin-susceptible Legionella spp. The superior potency of gemifloxacin compared with erythromycin indicates that it may be of use in the treatment of legionellosis. The significant PAE described here, combined with favourable pharmacokinetics, supports once-daily dosing for gemifloxacin in the treatment of legionella infections. PMID- 10824032 TI - In vitro activity of gemifloxacin and other antimicrobial agents against isolates of Bordetella pertussis and Bordetella parapertussis. AB - We investigated the activity of the novel quinolone agent gemifloxacin (SB 265805) and a panel of comparator agents against Bordetella pertussis and Bordetella parapertussis. Erythromycin, azithromycin, ciprofloxacin and gemifloxacin were consistently active against both species. An azithromycin- and erythromycin-resistant B. pertussis isolate was not resistant to any of the other agents tested (gemifloxacin MIC < or =0.008 mg/L; ciprofloxacin, 0.015 mg/L; ampicillin, 2.0 mg/L; trimethoprim-sulphamethoxazole, 4.0 mg/L). The potency of ampicillin, azithromycin, erythromycin, ciprofloxacin and trimethoprim sulphamethoxazole recorded against B. pertussis and B. parapertussis in this study was comparable to that noted in previous studies. However, MICs were generally higher than those noted in other trials; this may reflect the different methods used. Although in vitro data on the potency of gemifloxacin against B. pertussis and B. parapertussis have not previously been reported, these results are comparable to the potency of other quinolones against these pathogens. Should gemifloxacin achieve similar concentrations within the respiratory tract as other quinolones, this, coupled with its high in vitro potency, suggests that gemifloxacin has potential clinical efficacy in pertussis. PMID- 10824033 TI - In vitro activity of gemifloxacin compared with other antimicrobial agents against recent clinical isolates of streptococci. AB - This study investigated the in vitro activity of gemifloxacin (SB-265805) against 50 recent clinical isolates of Streptococcus pyogenes, Streptococcus agalactiae and viridans streptococci using the microdilution method. This activity was compared with that of the quinolone agents ofloxacin, ciprofloxacin, levofloxacin, trovafloxacin and grepafloxacin, and with that of penicillin, ampicillin, clarithromycin and azithromycin. Gemifloxacin was significantly more potent than the other quinolones tested. Its potency was equal to that of penicillin for S. agalactiae, and superior to that of penicillin for viridans streptococci. The MIC(50) of gemifloxacin for S. pyogenes (0.015 mg/L) was equal to that of penicillin, with an MIC(90) of 0.03 mg/L. Gemifloxacin was also active against isolates of S. agalactiae (MIC = 0.03-0.06 mg/L) and S. pyogenes (MIC = 0.03- 0.06 mg/L) with reduced susceptibility to ofloxacin (MIC = 4-8 mg/L) and grepafloxacin (MIC = 4 mg/L). These preliminary observations indicate that gemifloxacin is a promising antimicrobial agent for clinical use. PMID- 10824034 TI - Review of the in vitro activity of gemifloxacin against gram-positive and gram negative anaerobic pathogens. AB - Published reports on the in vitro activity of gemifloxacin mesylate (SB 265805), a new fluoronaphthyridone, against anaerobic pathogens are reviewed here. The studies used a variety of media, inocula and antimicrobial agents. Using a proposed breakpoint of 0.5 mg/L, these studies showed that gemifloxacin had generally higher potency against Gram-positive anaerobes (Clostridium perfringens, all Peptostreptococcus spp.) and fusobacteria (Fusobacterium nucleatum, Fusobacterium necrophorum) and moderate but variable potency against Gram-negative anaerobes. Bacteroides stercoris, Bacteroides tectum and many Bacteroides fragilis isolates were inhibited by concentrations of < or =0.5 mg/L, while the other species of the B. fragilis group required higher concentrations for inhibition. Species variability was evident: Porphyromonas asaccharolytica, Porphyromonas canoris, Porphyromonas gingivalis, Porphyromonas macaccae, Prevotella heparinolytica and Prevotella intermedia were susceptible to 0.5 mg/L of gemifloxacin while most other Porphyromonas and Prevotella spp. were not. These data suggest that gemifloxacin may have a clinical role in the treatment of certain dental, head and neck and pleuropulmonary infections in which Gram positive anaerobes, fusobacteria and some Prevotella and Porphyromonas spp. may predominate. PMID- 10824035 TI - Anti-gonococcal activity of gemifloxacin against fluoroquinolone-resistant strains and a comparison of agar dilution and Etest methods. AB - Gemifloxacin is a novel quinolone with excellent activity against Gram-positive and some Gram-negative pathogens. Its activity was tested against 150 Neisseria gonorrhoeae strains, including 50 ciprofloxacin-resistant isolates, using reference agar dilution and Etest methods. Gemifloxacin was found to be highly potent against ciprofloxacin-susceptible strains (MIC(90) 0.008 mg/L), but was 16 fold less potent against ciprofloxacin-resistant gonococci. The order of quinolone potency against these fluoroquinolone-resistant mutants was: gemifloxacin (MIC(90) 0.12 mg/L) > trovafloxacin (0.25 mg/L) > moxifloxacin = grepafloxacin (0.5 mg/L) > ciprofloxacin (1 mg/L). Etest and reference agar dilution MIC results showed excellent correlation (r = 0.96) and >98% of MICs were within +/-1 log(2) dilution step (essential agreement). The excellent potency of gemifloxacin indicates its potential for the treatment of infections with quinolone-resistant N. gonorrhoeae. PMID- 10824036 TI - Bactericidal and bacteriostatic activity of gemifloxacin against Acinetobacter spp. in vitro. AB - This study compared the in vitro bacteriostatic activity of gemifloxacin (SB 265805) and a panel of test antimicrobial agents against 100 clinical isolates of Acinetobacter spp. (47 Acinetobacter baumannii, 18 Acinetobacter anitratus, 18 Acinetobacter lwoffii, 13 Acinetobacter calcoaceticus and four other Acinetobacter spp.). Gemifloxacin (MIC(50/90) 0.06/16 mg/L) was more than eight fold more potent than ciprofloxacin (0.5/>128 mg/L), two- to eight-fold more potent than grepafloxacin, moxifloxacin, levofloxacin, ofloxacin and gatifloxacin, and of similar potency to trovafloxacin and sparfloxacin. Cross resistance was seen only within the quinolone group and did not extend to non quinolone antimicrobials. The bactericidal activities of gemifloxacin and the six comparator quinolones were investigated by dose-response and time-kill studies against A. baumannii ATCC 19606 at their optimum bactericidal concentration (OBC) and at 4 x MIC. At the OBC there was no significant difference between the quinolones, but at 4 x MIC gemifloxacin showed superior activity, reducing the viable count by almost 2 log(10) in 30 min compared with a 1 log(10) reduction seen with the other drugs. This enhanced killing extended over 24 h, reducing cell numbers by >4 log(10). These data suggest that gemifloxacin has the potential to be of therapeutic value in the treatment of infection by Acinetobacter spp. PMID- 10824037 TI - Comparative in vivo activity of gemifloxacin in a rat model of respiratory tract infection. AB - The in vivo efficacy of the novel quinolone gemifloxacin (SB-265805) was examined in a rat respiratory tract infection (RTI) model against four strains of Streptococcus pneumoniae and two strains of Haemophilus influenzae with varying susceptibilities to standard antimicrobial agents. Animals were infected intrabronchially to produce pneumonia and therapy with oral gemifloxacin, amoxycillin-clavulanate, ciprofloxacin, cefuroxime, azithromycin, trovafloxacin, grepafloxacin or levofloxacin was started 24 h after infection. The doses administered were chosen to approximate in the rat the serum or tissue concentrations measured in humans following therapeutic dosing. Therapy continued once- or twice-daily for 3 days, and approximately 17 h after the end of therapy the lungs were excised for bacterial enumeration. Following infection with strains of S. pneumoniae, gemifloxacin produced a 3-5 log reduction in bacterial numbers compared with untreated animals. Gemifloxacin was as effective as amoxycillin- clavulanate, and was as potent or more potent than all other comparators. Notably, the quinolone agents trovafloxacin, ciprofloxacin, grepafloxacin and levofloxacin were significantly less effective (P < 0.01) than gemifloxacin: these agents reduced bacterial numbers by < or =3 log compared with untreated animals. Gemifloxacin produced a marked response against H. influenzae infection, reducing bacterial numbers significantly (P < 0.01) compared with untreated controls. Gemifloxacin was significantly more potent than cefuroxime and azithromycin. None of the other comparator agents was more potent than gemifloxacin. The excellent efficacy seen in these experimental models of RTI with S. pneumoniae and H. influenzae confirms the in vitro activity of gemifloxacin against these organisms. This indicates that gemifloxacin may be of significant benefit in the treatment of RTI. PMID- 10824038 TI - Comparative efficacy of gemifloxacin in experimental models of pyelonephritis and wound infection. AB - Gemifloxacin (SB-265805) is a potent, novel fluoroquinolone with broad-spectrum antimicrobial activity. In this study, the efficacy of gemifloxacin was studied in experimental models of Gram-negative pyelonephritis (caused by Escherichia coli or Proteus mirabilis) and Gram-positive wound infection resulting from Streptococcus pyogenes, Staphylococcus epidermidis or Staphylococcus aureus. Gemifloxacin activity against these pathogens was compared with those of amoxycillin-clavulanate, ciprofloxacin, cefuroxime, azithromycin, trovafloxacin, grepafloxacin, levofloxacin and tosufloxacin. Oral treatment was initiated 1 h after infection and continued once or twice daily for 3 days. Around 17 h after the end of treatment, animals were killed and the infected kidneys or the skin around the wound site were excised for the enumeration of viable bacteria. In the pyelonephritis model (either microorganism), gemifloxacin reduced bacterial numbers significantly (P < 0.01) compared with no treatment. No comparator agent had a greater effect than gemifloxacin. Notably, grepafloxacin and azithromycin were significantly less effective (P < 0.01) than gemifloxacin against E. coli pyelonephritis, and amoxycillin-clavulanate, azithromycin and trovafloxacin were inferior (P < 0.01) against P. mirabilis infection. In the S. pyogenes wound infection model, gemifloxacin, amoxycillin-clavulanate, cefuroxime and azithromycin reduced bacterial numbers significantly compared with controls (P < 0.01). Results for the comparator quinolones were not significantly different from untreated controls (P > 0.05). Gemifloxacin was also effective against staphylococcal infection, as were grepafloxacin and levofloxacin, while ciprofloxacin, trovafloxacin and tosufloxacin were significantly less effective against these pathogens than gemifloxacin (P < 0.01). No comparator agent had greater activity than gemifloxacin against S. pyogenes or S. aureus infections. These data demonstrate the potential benefit of gemifloxacin in the treatment of Gram-negative urinary tract infection and Gram-positive skin and soft tissue infection. PMID- 10824039 TI - Efflux and target mutations as quinolone resistance mechanisms in clinical isolates of Streptococcus pneumoniae. AB - The aim of this study was to characterize quinolone resistance mechanisms in strains of Streptococcus pneumoniae with increased MICs of ofloxacin. These strains were also tested for their susceptibility to a battery of quinolone antimicrobial agents, including gemifloxacin. Of the S. pneumoniae isolates used, 27 were susceptible to ofloxacin, 18 intermediate and 48 resistant (ofloxacin MIC <4, 4 and >4 mg/L, respectively). In general, the ofloxacin-susceptible strains had no amino acid substitutions in GyrA, GyrB, ParC or ParE. Moderate increases in MIC were associated with substitutions in the quinolone resistance-determining region (QRDR) of ParC, while the highest MICs were found for strains that also had substitutions in the QRDR of GyrA. The most common substitutions were Ser79- >Phe in ParC and Ser81-->Phe in GyrA. Other substitutions were identified within the QRDR of ParC and outside the QRDR of ParC and ParE; these did not appear to affect susceptibility. The effects of antimicrobial efflux pumps were studied by determining MICs of a range of quinolones in the presence and absence of reserpine, an inhibitor of Gram-positive efflux pumps. Our results indicated that high-level resistance, caused entirely by efflux, was seen in a minority of ofloxacin-resistant S. pneumoniae strains. Testing the susceptibility of quinolone-resistant strains to gemifloxacin, ciprofloxacin, norfloxacin, ofloxacin and trovafloxacin revealed that gemifloxacin was least affected by this large variety of resistance mechanisms and was the only quinolone with MICs of < or =0.5 mg/L for all strains in this study. These results suggest that gemifloxacin is highly potent against S. pneumoniae and may also be effective against strains resistant to other quinolones. PMID- 10824040 TI - Purification of pneumococcal type II topoisomerases and inhibition by gemifloxacin and other quinolones. AB - Topoisomerase IV and DNA gyrase were purified from a ciprofloxacin-sensitive Streptococcus pneumoniae strain and from two clinical isolates of S. pneumoniae with high-level resistance to ciprofloxacin by means of a gene cloning method in Escherichia coli. All the quinolones tested (gemifloxacin, trovafloxacin, levofloxacin, ciprofloxacin and grepafloxacin) were able to inhibit topoisomerase IV at lower concentrations than those required for DNA gyrase, suggesting that topoisomerase IV is the primary target in the three pneumococci, in agreement with recently published enzyme data. Gemifloxacin (SB-265805) was found to be the most active agent against topoisomerase IV but, surprisingly, not against DNA gyrase. These findings indicate that the potent in vitro activity of gemifloxacin against S. pneumoniae, including ciprofloxacin-resistant strains, results from a strong affinity for pneumococcal topoisomerase IV. PMID- 10824041 TI - Bactericidal activity of gemifloxacin and other quinolones against Streptococcus pneumoniae. AB - This study compared the bactericidal activity of gemifloxacin (SB-265805) and a panel of test quinolones against two ciprofloxacin-resistant pneumococcal strains (Streptococcus pneumoniae 502226 and 503244) and one ciprofloxacin-sensitive strain (S. pneumoniae C3LN4). Activities were compared by calculating the bactericidal index of these agents. Gemifloxacin was found to be the most bactericidal quinolone tested against these strains. This finding confirms previous data indicating the superior in vitro activity of gemifloxacin against pneumococci, including ciprofloxacin-resistant strains. Although both ciprofloxacin-resistant strains tested had similar quinolone MICs, they differed considerably in their susceptibility to the bactericidal action of these agents. S. pneumoniae 502226 was more readily killed by quinolones than S. pneumoniae 503244 but, as would be expected, both were less susceptible than the ciprofloxacin-sensitive strain. Of the quinolones tested, trovafloxacin showed disproportionally poor activity against the ciprofloxacin-resistant strains even though potent activity was present against the ciprofloxacin-sensitive strain. These data highlight the importance of assessing quinolone bactericidal activity in addition to the MIC when evaluating new members of this antimicrobial class. PMID- 10824042 TI - Preface PMID- 10824043 TI - Chemotherapy for upper gastrointestinal tumours. AB - The aim of this review is to identify current chemotherapy treatment for tumours of the oesophagus, stomach, pancreas, and liver. The role of both neoadjuvant, adjuvant, and palliative chemotherapy regimens will be discussed. This review will be of interest to oncologists in clarifying current issues regarding chemotherapy, and to physicians in other medical specialties, to increase their general understanding of benefits and drawbacks of chemotherapy in this patient group. PMID- 10824044 TI - Advances in gastroenterology and hepatology. AB - This is a review of some of the most important growing points in the specialties of gastroenterology and hepatology. It does not aim to be completely comprehensive but to pick out major areas of importance to examination candidates and doctors without special experience in the field. Topics covered include: upper gastrointestinal haemorrhage; Barrett's oesophagus; carcinoma of the oesophagus; achalasia; Helicobacter pylori; duodenal ulcer prevention; coeliac disease; dermatitis herpetiformis; Crohn's disease; small bowel overgrowth; ulcerative colitis; carcinoma of the large bowel; obesity; endoscope sterilisation; gall stones; liver transplantation; autoimmune liver disease; viral hepatitis; metabolic liver diseases; and pancreatic insufficiency. PMID- 10824045 TI - Progressive supranuclear palsy. AB - Progressive supranuclear palsy (PSP) or Steele-Richardson-Olszewski syndrome is a neurodegenerative disease of middle and late age. It is under-diagnosed not only by general physicians but also by neurologists. The cause of PSP is not known. Exposure to toxins and viruses has been proposed in the aetiology of PSP without any concrete evidence. The features of PSP resemble those of Parkinson's disease and the two diseases are often confused. Corticobasal degeneration and multisystem atrophy are other differential diagnoses. Despite certain common features with Parkinson's disease, corticobasal degeneration, and mutisystem atrophy, there are important differences that help to differentiate it from these disorders. PMID- 10824046 TI - Permanent pacemaker insertion in a district general hospital: indications, patient characteristics, and complications. AB - This report reviews the experience of permanent pacemaker insertion in a district general hospital (catchment population of 350 000) and makes a comparison with the national database and other hospitals in the UK. METHODS: The records of all patients receiving a permanent pacemaker in the inclusive period January 1996 to December 1998 were reviewed. Data collected included number of patients paced each year, age, sex, indications, and complications. RESULTS: In the three years reviewed 200 patients received new permanent pacemakers, a rate of 190 per million population per year, which is similar to the national implantation rate of permanent pacemakers but lower than that of most European countries (see discussion). The majority of patients paced were elderly (75% were above the age of 70 years). Atrioventricular block (including complete heart block, 45%, and Mobitz type 2 block, 12.5%) was the commonest indication for permanent pacemaker insertion, followed by sick sinus syndrome (25%) and these findings are comparable to those reported previously. However, carotid sinus syndrome was responsible for 16% of the patients paced and this was higher than that reported in the national database (6.5%). Only 1% of the pacemaker modes used was inappropriate and the complication rate was low at 3%. CONCLUSIONS: This report confirms that permanent pacemaker insertion can be effectively and safely provided locally for the increasingly ageing population. The implantation rate both locally and nationally is still much lower than that of some countries in Europe. PMID- 10824047 TI - Trends in diagnostic and therapeutic criteria in Graves' disease in the last 10 years. AB - A questionnaire describing a typical clinical case of Graves' disease and 10 variations on it was mailed to 70 Spanish units of endocrinology with the aim of assessing the new diagnostic and therapeutic trends for hyperthyroidism caused by Graves' disease in Spain and to compare the results obtained from previous studies carried out in Europe and Spain 10 years previously. Responses indicated that thyrotrophin (98%) and free thyroxine (88%) were the most used tests in the in vitro diagnosis of Graves' disease with a significant decrease in the use of total thyroxine, total triiodothyronine, and thyroglobulin in comparison with the surveys conducted 10 years previously in Europe and Spain. The presence of antibodies against the thyrotrophin receptor was the most frequently used immune marker in the diagnosis (78%) and the new use of antithyroperoxidase antibodies (36%) in diagnosis is noteworthy. Antithyroid drugs remain the treatment of choice (98%). Surgery was used mainly for large size goitres (33%) and radioiodine for recurrences after medical (61%) or surgical (80%) treatment. In conclusion, the responses obtained from this questionnaire provide insight into current specialist diagnostic and therapeutic practices with respect to Graves' disease and which could be of value to non-specialist units of endocrinology. PMID- 10824048 TI - Status of antithyroid antibodies in Bangladesh. AB - To study autoimmunity among thyroid diseases, 397 thyroid patients (age 30 (13) years; M/F 75/322) from two referral centres in Bangladesh and 94 healthy controls (age 30 (13) years; M/F 24/70) were studied for antimicrosomal and antithyroglobulin antibodies. Thyroid patients were clinically grouped as suspected autoimmune thyroid disease (AITD), non-autoimmune, or indeterminate groups (where no decision could be reached). Antimicrosomal antibody was strongly positive in 19.4% and weakly positive in 7.3% of patients but only 4.3% and 2.1% respectively in the controls (chi(2) = 17.852; p = 0.000) whereas strong and weak positivity were 27.2% and 6. 8% in patients compared with 8.5% and 4.3% respectively in the controls (chi(2) = 16.916; p = 0.000) for antithyroglobulin antibody. Antibodies were positive in 63.0% with Hashimoto's thyroiditis, 36.4% with Graves' disease, and 44.7% with atrophic thyroiditis among the autoimmune group. In the non-autoimmune group antibodies were positive in 100% with multinodular hypothyroidism, 46.7% with subacute thyroiditis, 40.0% with suspected iodine deficiency goitre, 31.3% with toxic multinodular goitre, 30.8% with non-toxic solitary nodules, and 19.4% with simple diffuse goitre. None was positive for antimicrosomal antibody without being positive for antithyroglobulin antibody. The two antibodies strongly correlated in both patients (r = 0.977, p = 0.000) and controls (r = 0.986, p = 0.000). About 9% (36/397) of patients were mismatched with the final diagnosis on antibody measurement; most of them had Hashimoto's thyroiditis (33/36). Prevalence of AITD among thyroid patients was 48.36%. Specificity of antimicrosomal and antithyroglobulin antibodies were 93% and 87%. It was concluded that AITD is not uncommon in Bangladesh; antimicrosomal antibody is a useful marker for AITD and unless antibodies are checked, an appreciable number of patients with AITDs will remain undetected. PMID- 10824049 TI - Evaluation of amlodipine, lisinopril, and a combination in the treatment of essential hypertension. AB - Angiotensin converting enzyme (ACE) inhibitors and dihydropyridine calcium antagonists are well established and widely used as monotherapy in patients with mild to moderate essential hypertension. Earlier studies combining short acting drugs from these classes require multiple dosing and were associated with poor compliance. Availability of longer acting compounds allows once daily administration to avoid the inconvenience of a multiple daily dose. It was decided to perform a randomised double blind, crossover study with the long acting calcium channel blocker amlodipine and the long acting ACE inhibitor lisinopril, given either alone or in combination in essential hypertension. Twenty four patients with diastolic blood pressure (DBP) between 95 and 104 mm Hg received amlodipine 2.5 mg and 5 mg, lisinopril 5 mg and 10 mg, and their combination as per a prior randomisation schedule. Supine and standing blood pressure and heart rate were recorded at weekly intervals. Higher doses of both the drugs individually or in combination were used if the target supine DBP below 90 mm Hg was not achieved. There was a significant additional blood pressure lowering effect with the combination when compared either with amlodipine or lisinopril alone. Five mg amlodipine and 10 mg lisinopril monotherapy achieved the target blood pressure in 71% and 72% patients respectively. The combination of 2.5 mg amlodipine with 5 mg lisinopril produced a much more significant lowering of blood pressure in a higher percentage of patients than that with an individual low dose. PMID- 10824050 TI - Use of a computerised maternity information system to improve clinical effectiveness: thromboprophylaxis at caesarean section. AB - An audit of the introduction of a protocol for thromboprophylaxis at caesarean section revealed over treatment of low risk women and the under treatment of high risk women. A routine computer generated risk assessment profile was introduced as part of a maternity information system. Reaudit showed a significant improvement in adherence to the thromboprophylaxis protocol in all risk groups. PMID- 10824051 TI - Teaching medical undergraduates basic clinical skills in hospice--is it practical? AB - AIM: Basic clinical skills teaching to medical undergraduates was a new departure for the hospice stimulated by the changing philosophies and organisation of students' training. This study was undertaken to assess the practicalities of the venture. METHOD: Questionnaires were designed for each of the three major groups of people involved, namely the students, the patients, and the hospice nurses. Involved patients completed theirs after teaching sessions, while the students and nurses were given two different questionnaires each, one at the start and another at the end of the academic year. RESULTS: All students completed both questionnaires. Overall they had acquired adequate skills to pass their end of year assessments and considered themselves more comfortable with difficult situations than may otherwise have been the case. The majority of patients had enjoyed the experience and found it personally educational and a change to hospice routines. The nurses' response rates were very poor, limiting any conclusions that could be drawn. CONCLUSION: The venture was successful, stimulating, and practical for patients and students. Its impact on the nurses remains uncertain but, by their unusual lack of opinion expression, it can be inferred tentatively that this was minimal. PMID- 10824052 TI - Hypersensitivity syndrome caused by amitriptyline administration. AB - Adverse cutaneous manifestations are among the most common side effects associated with psychotropic drugs. Skin reactions due to amitriptyline (a tricyclic antidepressant agent) include rashes and hypersensitivity reactions (for example, urticaria and photosensitivity) as well as hyperpigmentation. Hypersensitivity syndrome is a specific severe idiosyncratic reaction causing skin, liver, joint, and haematological abnormalities, which usually resolve after the discontinuation of the implicated drug. A case of a 24 year old woman who experienced hypersensitivity syndrome three weeks after the initiation of amitriptyline is reported. PMID- 10824053 TI - Primary amyloidosis of the larynx. AB - Primary laryngeal amyloidosis is a rare benign disease of unknown aetiology. It can present with dysphonia or stridor. A woman presenting with airway compromise, who required a tracheostomy, is reported. PMID- 10824054 TI - Extensive psoriasis induced by interferon alfa treatment for chronic hepatitis C. AB - A 47 year old man with chronic hepatitis C was treated with interferon alfa, 3 million units three times a week, and developed widespread plaque psoriasis within weeks of starting interferon therapy. There was no previous history of psoriasis. The psoriasis was characterised by extensive nail involvement and plaques at the interferon injection sites. The patient relapsed after a total of 12 months of interferon and was subsequently treated with interferon and tribavirin (ribavirin) with recurrence of the psoriasis. PMID- 10824055 TI - Histoplasmosis of the small bowel in patients with AIDS. AB - Two cases of jejunal strictures caused by Histoplasma capsulatum in AIDS patients are presented. Both patients were intravenous drug abusers. One patient, who was being treated for Pneumocystis carnii pneumonia, presented with jejunal perforation and the other presented with lower gastrointestinal bleeding and intestinal obstruction. On exploration, both patients were found to have jejunal strictures; one had intestinal perforation, and the other had intestinal obstruction with ulcers and strictures resulting in gastrointestinal bleeding. In areas where it is endemic, histoplasmosis is rarely disseminated. Dissemination is most commonly seen in immunosuppressed patients. Dissemination and extrapulmonary histoplasmosis is now included in the case definition of AIDS. PMID- 10824056 TI - Pseudovitamin D deficiency rickets--a report from the Indian subcontinent. AB - Pseudovitamin D deficiency rickets (also called vitamin D dependent rickets type I) is one of the types of inherited rickets and is caused by a deficit in renal 25-hydroxyvitamin D 1alpha-hydroxylase. This form of rickets has not been reported from the Indian subcontinent. Three patients with this disorder are presented. These patients were all females aged 3-20 years and presented with growth failure and skeletal deformities. All had florid clinical and radiological rickets. The biochemical abnormalities seen included hypocalcaemia, hypophosphataemia, and hyperphosphatasia. All patients had grossly raised 25 hydroxyvitamin D concentrations and markedly low to undetectable concentrations of 1,25-dihydroxyvitamin D. A disturbing feature of this study was the late referral of the patients. PMID- 10824057 TI - An unusual presentation of a common disorder. PMID- 10824058 TI - Young male with recurrent flaccid quadriparesis and complete recovery. PMID- 10824059 TI - Swelling in the caesarean section scar. PMID- 10824060 TI - An unusual case of chorea gravidarum. PMID- 10824062 TI - An unusual presentation of a common disorder PMID- 10824063 TI - Young male with recurrent flaccid quadriparesis and complete recovery PMID- 10824061 TI - Nausea and vomiting, a cause for concern? PMID- 10824065 TI - An unusual case of chorea gravidarum PMID- 10824064 TI - Swelling in the caesarean section scar PMID- 10824066 TI - Nausea and vomiting, a cause for concern? PMID- 10824067 TI - Anesthesia and transplantation PMID- 10824068 TI - Instant pharmacology PMID- 10824069 TI - Drugs in anaesthetic & intensive care practice PMID- 10824070 TI - Fundamentals of cardiovascular pharmacology PMID- 10824071 TI - Cardiovascular therapeutics PMID- 10824072 TI - Cardiopulmonary resuscitation by chest compression alone or with mouth-to-mouth ventilation. AB - BACKGROUND: Despite extensive training of citizens of Seattle in cardiopulmonary resuscitation (CPR), bystanders do not perform CPR in almost half of witnessed cardiac arrests. Instructions in chest compression plus mouth-to-mouth ventilation given by dispatchers over the telephone can require 2.4 minutes. In experimental studies, chest compression alone is associated with survival rates similar to those with chest compression plus mouth-to-mouth ventilation. We conducted a randomized study to compare CPR by chest compression alone with CPR by chest compression plus mouth-to-mouth ventilation. METHODS: The setting of the trial was an urban, fire-department-based, emergency-medical-care system with central dispatching. In a randomized manner, telephone dispatchers gave bystanders at the scene of apparent cardiac arrest instructions in either chest compression alone or chest compression plus mouth-to-mouth ventilation. The primary end point was survival to hospital discharge. RESULTS: Data were analyzed for 241 patients randomly assigned to receive chest compression alone and 279 assigned to chest compression plus mouth-to-mouth ventilation. Complete instructions were delivered in 62 percent of episodes for the group receiving chest compression plus mouth-to-mouth ventilation and 81 percent of episodes for the group receiving chest compression alone (P=0.005). Instructions for compression required 1.4 minutes less to complete than instructions for compression plus mouth-to-mouth ventilation. Survival to hospital discharge was better among patients assigned to chest compression alone than among those assigned to chest compression plus mouth-to-mouth ventilation (14.6 percent vs. 10.4 percent), but the difference was not statistically significant (P=0.18). CONCLUSIONS: The outcome after CPR with chest compression alone is similar to that after chest compression with mouth-to-mouth ventilation, and chest compression alone may be the preferred approach for bystanders inexperienced in CPR. PMID- 10824073 TI - Dexamethasone alone or in combination with ondansetron for the prevention of delayed nausea and vomiting induced by chemotherapy. AB - BACKGROUND: The prevention of delayed nausea and vomiting caused by moderately emetogenic chemotherapy for cancer has not been studied systematically. METHODS: We enrolled patients who were scheduled to receive chemotherapy for the first time in a double-blind, randomized, multicenter study. All the patients received ondansetron combined with dexamethasone for prophylaxis against emesis that might occur within 24 hours after the start of chemotherapy (acute emesis). They were then divided into two groups: patients who did not have either vomiting or moderate-to-severe nausea (the low-risk group) and patients who had one or both (the high-risk group). Patients in the low-risk group were then randomly assigned to one of the following regimens, given on days 2 through 5 after the start of chemotherapy: oral placebo, 4 mg of dexamethasone given orally twice daily, or 8 mg of ondansetron in combination with 4 mg of dexamethasone, given orally twice daily. Patients in the high-risk group were randomly assigned to receive oral dexamethasone alone or in combination with ondansetron at the same doses as those used in the low-risk group. RESULTS: Among the 618 patients in the low-risk group, there was a complete absence of both delayed vomiting and moderate-to severe nausea in 91.8 percent of those who received ondansetron combined with dexamethasone, 87.4 percent of those who received dexamethasone alone, and 76.8 percent of those who received placebo. The proportions of patients who were protected by dexamethasone combined with ondansetron or by dexamethasone alone were significantly greater than the proportion protected by placebo (P<0.001 and P<0.02, respectively). Of the 87 patients in the high-risk group, complete protection was achieved in 40.9 percent of those treated with ondansetron and dexamethasone and in 23.3 percent treated with dexamethasone alone (P not significant). CONCLUSIONS: The best way to prevent delayed nausea and vomiting in patients receiving moderately emetogenic chemotherapy is to control these complications within the first 24 hours after the start of chemotherapy. Dexamethasone alone provides adequate protection against delayed emesis in patients at low risk (those who have not had acute emesis). PMID- 10824074 TI - Association between early-onset Parkinson's disease and mutations in the parkin gene. AB - BACKGROUND: Mutations in the parkin gene have recently been identified in patients with early-onset Parkinson's disease, but the frequency of the mutations and the associated phenotype have not been assessed in a large series of patients. METHODS: We studied 73 families in which at least one of the affected family members was affected at or before the age of 45 years and had parents who were not affected, as well as 100 patients with isolated Parkinson's disease that began at or before the age of 45 years. All subjects were screened for mutations in the parkin gene with use of a semiquantitative polymerase-chain-reaction assay that simultaneously amplified several exons. We sequenced the coding exons in a subgroup of patients. We also compared the clinical features of patients with parkin mutations and those without mutations. RESULTS: Among the families with early-onset Parkinson's disease, 36 (49 percent) had parkin mutations. The age at onset ranged from 7 to 58 years. Among the patients with isolated Parkinson's disease, mutations were detected in 10 of 13 patients (77 percent) with an age at onset of 20 years or younger, but in only 2 of 64 patients (3 percent) with an age at onset of more than 30 years. The mean (+/-SD) age at onset in the patients with parkin mutations was younger than that in those without mutations (32+/-11 vs. 42+/-11 years, P<0.001), and they were more likely to have symmetric involvement and dystonia at onset, to have hyperreflexia at onset or later, to have a good response to levodopa therapy, and to have levodopa-induced dyskinesias during treatment. Nineteen different rearrangements of exons (deletions and multiplications) and 16 different point mutations were detected. CONCLUSIONS: Mutations in the parkin gene are a major cause of early-onset autosomal recessive familial Parkinson's disease and isolated juvenile-onset Parkinson's disease (at or before the age of 20 years). Accurate diagnosis of these cases cannot be based only on the clinical manifestations of the disease. PMID- 10824075 TI - Eosinophilia with aberrant T cells and elevated serum levels of interleukin-2 and interleukin-15. PMID- 10824076 TI - Images in clinical medicine. Xanthogranulomatous pyelonephritis. PMID- 10824077 TI - The volume of primary angioplasty procedures and survival after acute myocardial infarction. National Registry of Myocardial Infarction 2 Investigators. AB - BACKGROUND: There is an inverse relation between mortality from cardiovascular causes and the number of elective cardiac procedures (coronary angioplasty, stenting, or coronary bypass surgery) performed by individual practitioners or hospitals. However, it is not known whether patients with acute myocardial infarction fare better at centers where more patients undergo primary angioplasty or thrombolytic therapy than at centers with lower volumes. METHODS: We analyzed data from the National Registry of Myocardial Infarction to determine the relation between the number of patients receiving reperfusion therapy (primary angioplasty or thrombolytic therapy) and subsequent in-hospital mortality. A total of 450 hospitals were divided into quartiles according to the volume of primary angioplasty. Multiple logistic-regression models were used to determine whether the volume of primary angioplasty procedures was an independent predictor of in-hospital mortality among patients undergoing this procedure. Similar analyses were performed for patients receiving thrombolytic therapy at 516 hospitals. RESULTS: In-hospital mortality was 28 percent lower among patients who underwent primary angioplasty at hospitals with the highest volume than among those who underwent angioplasty at hospitals with the lowest volume (adjusted relative risk, 0.72; 95 percent confidence interval, 0.60 to 0.87; P<0.001). This lower rate, which represented 2.0 fewer deaths per 100 patients treated, was independent of the total volume of patients with myocardial infarction at each hospital, year of admission, and use or nonuse of adjunctive pharmacologic therapies. There was no significant relation between the volume of thrombolytic interventions and in-hospital mortality among patients who received thrombolytic therapy (7.0 percent for patients in the highest-volume hospitals vs. 6.9 percent for those in the lowest-volume hospitals, P=0.36). CONCLUSIONS: Among hospitals in the United States that have full interventional capabilities, a higher volume of angioplasty procedures is associated with a lower mortality rate among patients undergoing primary angioplasty, but there is no association between volume and mortality for thrombolytic therapy. PMID- 10824078 TI - Hyponatremia. PMID- 10824079 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 16-2000. A 53-year-old woman with swelling of the right breast and bilateral lymphadenopathy. PMID- 10824080 TI - Cardiopulmonary resuscitation--strengthening the links in the chain of survival. PMID- 10824081 TI - Nonaccredited medical education in the United States. PMID- 10824082 TI - Correction: Retroperitoneal Hemorrhage. PMID- 10824083 TI - Correction: Case Records of the Massachusetts General Hospital (Case 9-2000). PMID- 10824084 TI - Evolutionary analysis of the ErbB receptor and ligand families. AB - We have compared all available deduced protein sequences of the ErbB family of receptors and their ligands. Analysis of the aligned sequences of the receptors indicates that there are some differences in the receptors that are specific to invertebrates. In addition, comparison of the vertebrate ErbB receptors suggest that a gene duplication event generated two ancestral receptors, the ErbB3/ErbB4 precursor and the ErbB1/ErbB2 precursor. Subsequent gene duplications of these precursors generated the four receptors present in mammals. Analysis of the sequences for the known ligands of the ErbB receptors suggests that the vertebrate ligands segregate into the ErbB1 ligands and the ErbB3/ErbB4 ligands, paralleling the evolution of the receptors; however, it is difficult to ascertain any correlation between the invertebrate and the vertebrate ligands. Even though ErbB3 is kinase-impaired, there is significant conservation of the kinase domain within the vertebrate lineage (human, rat, and F. rubripes), suggesting some function for this domain other than kinase activity, such as mediating protein protein interactions that are involved in receptor dimerization and/or activation of the kinase domain of the heterodimerization partner. To date, no ligand for ErbB2 has been identified, and comparison of the extracellular domains of ErbB2 reveals two regions that are not conserved across the mammalian species. These two regions of divergence align with sequences in ErbB1 that have been shown to be proximal to the amino-terminus and to the carboxyl-terminal region, respectively, of bound EGF. Further, one of these regions contains an insertion, relative to the other members of the mammalian ErbB family, which might affect the ligand binding site and provide a structural basis for this receptor's apparent inability to bind ligand independently. PMID- 10824085 TI - Duplication and quadruplication of Arabidopsis thaliana cysteinyl- and asparaginyl-tRNA synthetase genes of organellar origin. AB - Two cysteinyl-tRNA synthetases (CysRS) and four asparaginyl-tRNA synthetases (AsnRS) from Arabidopsis thaliana were characterized from genome sequence data, EST sequences, and RACE sequences. For one CysRS and one AsnRS, sequence alignments and prediction programs suggested the presence of an N-terminal organellar targeting peptide. Transient expression of these putative targeting sequences joined to jellyfish green fluorescent protein (GFP) demonstrated that both presequences can efficiently dual-target GFP to mitochondria and plastids. The other CysRS and AsnRSs lack targeting sequences and presumably aminoacylate cytosolic tRNAs. Phylogenetic analysis suggests that the four AsnRSs evolved by repeated duplication of a gene transferred from an ancestral plastid and that the CysRSs also arose by duplication of a transferred organelle gene (possibly mitochondrial). These case histories are the best examples to date of capture of organellar aminoacyl-tRNA synthetases by the cytosolic protein synthesis machinery. PMID- 10824086 TI - Conserved structural features in class I major fimbrial subunits (Pilin) in gram negative bacteria. Molecular basis of classification in seven subfamilies and identification of intrasubfamily sequence signature motifs which might Be implicated in quaternary structure. AB - Type 1 and P-pili are prototype members of Class I fimbriae produced by Gram negative bacteria. Despite common structural characteristics, the low level of amino acid sequence conservation among the Class I major fimbrial subunits (pilins) indicates considerable evolutionary distance between members of this superfamily. We highlight here structural relatedness between Class I pilins from their two-dimensional sequence analysis using hydrophobic cluster analysis (HCA) and secondary structure predictions (PHD program). We present evidence that all members of the Class I pilin family have clear structural relatedness and suggest that classification based on phylogenetic analysis of Class I pilins into seven subfamilies correlates with differences in structural properties of the amino acid sequences. Using a sensitive alignment process (HCA), we identified 29 residues in topohydrophobic positions which probably play a prominent role in folding. The most striking aspects that distinguish the different pilin subfamilies are (i) large variation in the length of the loops connecting the structurally conserved regions and (ii) intrasubfamily sequence signature motifs located on regions predicted to be in the beta-conformation. We suggest that these "intrasubfamily sequence signature motifs" are part of interactive surfaces which participate in subunit-subunit interactions. These motifs prove highly useful in characterizing and classifying new Class I fimbriae that have not yet been described and whose sequence diverges appreciably from those of characterized groups. (After the submission of our manuscript, the experimental structure of Class I pilus subunits was published. In light of these actual pilin structures, a comparison has been made between the predicted results and the crystal structure in the Note Added in Proof.) PMID- 10824087 TI - Male-biased mutation rates revealed from Z and W chromosome-linked ATP synthase alpha-subunit (ATP5A1) sequences in birds. AB - Whether the mutation rate differs between sexes has been a matter of discussion for years. Molecular analyses of mammals have indicated that males mutate more often than females, as manifested by the faster rate of neutral sequence evolution on the Y chromosome than on the X chromosome. However, these observations can as well be interpreted as specific reduction of the X chromosome mutation rate, which would be adaptive because of reducing the number of slightly deleterious recessive mutations exposed in hemizygote males. Recently, data from birds have suggested that vertebrate mutation rates may indeed be male-biased. In birds, females are the heterogametic sex (ZW), and analyses of the Z-linked CHD1Z gene have shown that it evolves faster than its W-linked and thus female-specific homologue, CHD1W. We have now studied the second avian gene known to exist in a copy on the nonrecombining regions of both the Z and the W chromosome, viz., the ATP synthase alpha-subunit (ATP5A1). In independent comparisons of three pairs of bird species from divergent lineages, intron sequences of the Z-linked copy (ATP5A1Z) were consistently found to evolve faster than the W-linked copy (ATP5A1W). From these data we calculated male-to-female mutation rate ratios (alpha) of 1.8, 2.3, and 5.0 in Galliform, Anseriform, and Ciconiiform lineages, respectively. Therefore, this study provides independent support for a male biased mutation rate in birds. PMID- 10824088 TI - Evolutionary dynamics of HTLV-I. AB - Using mathematical models to describe the in vivo dynamics of HTLV-I infection, an explanation is offered for the slow rate of evolution of HTLV-I relative to HIV-1. In agreement with experimental findings, it is assumed that cell activation is required for successful replication in T helper cells and that HTLV I induces a significant degree of bystander activation. It is found that the rate of evolution of HTLV-I is limited by the restricted availability of activated uninfected T cells, both at high and low proviral loads. This limits the within host sequence diversity of HTLV-I and may therefore account for the slow rate of evolution of the virus in the population. Specific differences in the in vivo dynamics of HTLV-I and HIV-1 are identified which may account for the discrepancy in the rate of evolution of these two retroviruses. PMID- 10824089 TI - The vertebrate 7S K RNA separates hagfish (Myxine glutinosa) and lamprey (Lampetra fluviatilis). AB - 7S RNA sequences from the hagfish (Myxiniformes) and lamprey (Petromyzontiformes) were cloned and analyzed. In both species, 7S L RNA (also designated SRP RNA, since it represents the RNA constituent of the signal recognition particle) was clearly detectable. The sequence similarity between the two species was 86%, compared with about 75% similarity between either of these species and mammals. 7S K RNA was also cloned from the lamprey. The similarity between the 7S K RNA of the lamprey and that of mammals was 68%. Interestingly, several interspersed elements were found with nearly 100% similarity compared with mammals. In contrast to the lamprey, no 7S K RNA-related sequences were detectable among hagfish RNA, neither in northern blots nor with the PCR assay. In view of the significant conservation between the 7S K RNA of lamprey and that of mammals (human), this unexpected result clearly separates lamprey and hagfish. In addition, the lack of detectable 7S K RNA sequences in an outgroup, such as amphioxus, indicates that these results do not reflect an autapomorphy of hagfish. Therefore, our data provide additional support to the notion of a sister group relationship between Petromyzontiformes and gnathostomous vertebrates to the exclusion of Myxiniformes. PMID- 10824090 TI - Episodic evolution of protein hormones: molecular evolution of pituitary prolactin. AB - Previous studies have shown that pituitary growth hormone displays an episodic pattern of evolution, with a slow underlying evolutionary rate and occasional sustained bursts of rapid change. The present study establishes that pituitary prolactin shows a similar pattern. During much of tetrapod evolution the sequence of prolactin has been strongly conserved, showing a slow basal rate of change (approx 0.27x10(9) substitutions/amino acid site/year). This rate has increased substantially ( approximately 12- to 38-fold) on at least four occasions during eutherian evolution, during the evolution of primates, artiodactyls, rodents, and elephants. That these increases are real and not a consequence of inadvertant comparison of paralogous genes is shown (for at least the first three groups) by the fact that they are confined to mature protein coding sequence and not apparent in sequences coding for signal peptides or when synonymous substitutions are examined. Sequences of teleost prolactins differ markedly from those of tetrapods and lungfish, but during the course of teleost evolution the rate of change of prolactin has been less variable than that of growth hormone. It is concluded that the evolutionary pattern seen for prolactin shows long periods of near-stasis interrupted by occasional bursts of rapid change, resembling the pattern seen for growth hormone in general but not in detail. The most likely basis for these bursts appears to be adaptive evolution though the biological changes involved are relatively small. PMID- 10824091 TI - Divergence of noncoding sequences and of insertions encoding nonglobular domains at a genomic region well conserved in plasmodia. AB - To identify conserved features in the rapidly diverging portions of a well conserved locus, completely sequenced in Plasmodium falciparum and Plasmodium berghei, a computational method based on recurrence analysis was exploited. At the level of the genomic sequence, in both species, introns and intergenic sequences-though subject to rapid diversification-do not drift without constraints, but rather coevolve, in the sense that they maintain not only an AT rich base composition, but also a consistent use of recurring (AT)(n) tracts. One of the two genes present in the conserved locus encodes a protein that exhibits blocks of high similarity to the first enzyme in glutathione biosynthesis (gamma glutamylcysteine synthetase) but bears long low-complexity insertions, absent in other organisms. From an analysis of the aminoacid sequence, different constraints appear to act on the borders and on the central part of the insertions. Albeit maintaining a strong bias toward hydrophilic residues, central portions diverge more rapidly than borders, through point mutation and differential presence of entire tracts. PMID- 10824092 TI - The genotypic landscape during in vitro evolution of a catalytic RNA: implications for phenotypic buffering. AB - The Tetrahymena group I ribozyme catalyzes the cleavage of a phosphodiester linkage in specific sequences of RNA. This phenotype can be used in an in vitro selection-amplification process to evolve variants that are capable of RNA catalysis in the presence of Ca(2+) as the sole available cation. With sufficient genotypic characterization of the population as it evolves, we have a rare opportunity of observing how the information stored in an evolving population responds to selective pressures, such as the requisite of catalyzing RNA cleavage in the absence of Mg(2+) or Mn(2+). In the present work, we examine the population dynamics of this system using sequence information from previous experimental work. We focus on two issues: How does the information content of the population evolve? and Is the system evolving as an adaptive walk on a rugged landscape? To investigate these questions, information theoretical parameters are examined. The evolution of the population is visualized by mapping the genotypic frequency distribution onto a two-dimensional projection of sequence space. The projection was generated using Hamming distances from the wild-type, starting sequence and a catalytically successful, evolved sequence. The evolution of the information content of the system was measured by calculating the grammar complexity of the observed sequences, which showed a very slight increase over 12 generations. This result is consistent with the system performing a search for a local optimum. The dynamics of the population in this sequence space is consistent with an adaptive walk on an uncorrelated, or "rugged," genotypic landscape, despite the observation that the phenotypic progress of the population appears smooth. The relative insensitivity of the phenotypic landscape to the variegation of the genotypic landscape suggests that the former is buffered against variation in the latter through various epigenetic-like mechanisms. PMID- 10824093 TI - Identification of sequences encoding the detoxification metalloisomerase glyoxalase I in microbial genomes from several pathogenic organisms. AB - The ubiquitous glyoxalase system, which is composed of two enzymes, removes cellular cytotoxic methylglyoxal (MG). In an effort to identify critical residues conserved in the evolution of the first enzyme in this system, glyoxalase I (GlxI), as well as the structural implications of sequence alterations in this enzyme, a search of the National Center for Biotechnology Information (NCBI) database of unfinished genomes was undertaken. Eleven putative GlxI sequences from pathogenic organisms were identified and analyses of these sequences in relation to the known and previously identified GlxI enzymes were performed. Several of these sequences show a very high similarity to the Escherichia coli GlxI sequence, most notably the 79% identity of the sequence identified from Yersinia pestis, the causative agent of bubonic plague. In addition to the conservation of residues critical to binding the catalytic metal in all of the proposed GlxI enzymes, four regions in the Homo sapiens GlxI enzyme are absent in all of the bacterial GlxI sequences, with the exception of Pseudomonas putida. Removal of these regions may alter the active-site conformation of the bacterial enzymes in relation to that of the H. sapiens. These differences may be targeted for the development of inhibitors selective to the bacterial enzymes. PMID- 10824094 TI - Site-directed mutation of noncatalytic residues of Thermobifida fusca exocellulase Cel6B. AB - Fifteen mutant genes in six loop residues and eight mutant genes in five conserved noncatalytic active site residues of Thermobifida fusca Cel6B were constructed, cloned and expressed in Escherichia coli or Streptomyces lividans. The mutant enzymes were assayed for catalytic activity on carboxymethyl cellulose (CMC), swollen cellulose (SC), filter paper (FP), and bacterial microcrystalline cellulose (BMCC) as well as cellotetraose, cellopentaose, and 2, 4-dinitrophenyl beta-D-cellobioside. They were also assayed for ligand binding, enzyme processivity, thermostability, and cellobiose feedback inhibition. Two double Cys mutations that formed disulfide bonds across two tunnel forming loops were found to significantly weaken binding to ligands, lower all activities, and processivity, demonstrating that the movement of these loops is important but not essential for Cel6B function. Two single mutant enzymes, G234S and G284P, had higher activity on SC and FP, and the double mutant enzyme had threefold and twofold higher activity on these substrates, respectively. However, synergism with endocellulase T. fusca Cel5A was not increased with these mutant enzymes. All mutant enzymes with lower activity on filter paper, BMCC, and SC had lower processivity. This trend was not true for CMC, suggesting that processivity in Cel6B is a key factor in the hydrolysis of insoluble and crystalline cellulose. Three mutations (E495D, H326A and W329C) located near putative glycosyl substrate subsites -2, +1 and +2, were found to significantly increase resistance to cellobiose feedback inhibition. Both the A229V and L230C mutations specifically decreased activity on BMCC, suggesting that BMCC hydrolysis has a different rate limiting step than the other substrates. Most of the mutant enzymes had reduced thermostability although Cel6B G234S maintained wild-type thermostability. The properties of the different mutant enzymes provide insight into the catalytic mechanism of Cel6B. PMID- 10824095 TI - Formation of a native-like beta-hairpin finger structure of a peptide from the extended PDZ domain of neuronal nitric oxide synthase in aqueous solution. AB - Neuronal nitric oxide synthase (nNOS) is targeted to the cell membrane via interactions of its extended PDZ domain with PDZ domains of membrane-associated proteins including PSD-95 and alpha1-syntrophin. The formation of heterodimers between the nNOS PDZ domain and the PDZ domains of nNOS-binding proteins requires a stretch of continuous amino-acid residues C-terminal to the canonical nNOS PDZ domain. In this work, we show that a 27-residue peptide comprising the C-terminal extension of the extended nNOS PDZ domain is capable of binding to PSD-95. The structure of the 27-residue peptide in aqueous solution was determined using multidimensional NMR-spectroscopic techniques. The free peptide adopts a native like beta-hairpin finger structure in aqueous solution. The results indicate that the C-terminal extension peptide of the nNOS PDZ domain may represent a relatively independent structural unit in the mediation of the interaction between nNOS and PDZ domain-containing proteins including PSD-95 and alpha1 syntrophin. PMID- 10824096 TI - Role of ligand substitution on long-range electron transfer in azurins. AB - Azurin contains two potential redox sites, a copper centre and, at the opposite end of the molecule, a cystine disulfide (RSSR). Intramolecular electron transfer between a pulse radiolytically produced RSSR- radical anion and the blue Cu(II) ion was studied in a series of azurins in which single-site mutations were introduced into the copper ligand sphere. In the Met121His mutant, the rate constant for intramolecular electron transfer is half that of the corresponding wild-type azurin. In the His46Gly and His117Gly mutants, a water molecule is co ordinated to the copper ion when no external ligands are added. Both these mutants also exhibit slower intramolecular electron transfer than the corresponding wild-type azurin. However, for the His117Gly mutant in the presence of excess imidazole, an azurin-imidazole complex is formed and the intramolecular electron-transfer rate increases considerably, becoming threefold faster than that observed in the native protein. Activation parameters for all these electron transfer processes were determined and combined with data from earlier studies on intramolecular electron transfer in wild-type and single-site-mutated azurins. A linear relationship between activation enthalpy and activation entropy was observed. These results are discussed in terms of reorganization energies, driving force and possible electron-transfer pathways. PMID- 10824097 TI - N-carboxymethanofuran (carbamate) formation from methanofuran and CO2 in methanogenic archaea. Thermodynamics and kinetics of the spontaneous reaction. AB - N-carboxymethanofuran (carbamate) formation from unprotonated methanofuran (MFR) and CO2 is the first reaction in the reduction of CO2 to methane in methanogenic archaea. The reaction proceeds spontaneously. We address here the question whether the rate of spontaneous carbamate formation is high enough to account for the observed rate of methanogenesis from CO2. The rates of carbamate formation (v1) and cleavage (v2) were determined under equilibrium conditions via 2D proton exchange NMR spectroscopy (EXSY). At pH 7.0 and 300 K the second order rate constant k1* of carbamate formation from 'MFR'(MFR + MFRH+) and 'CO2' (CO2 + H2CO3 + HCO3-+ CO32-) was found to be 7 M-1.s-1 (v1 = k1* ['MFR'] ['CO2']) while the pseudo first order rate constant k2* of carbamate cleavage was 12 s-1 (v2 = k2* [carbamate]). The equilibrium constant K* = k1*/k2* = [carbamate]/['MFR']['CO2'] was 0.6 M-1 at pH 7.0 corresponding to a free energy change DeltaG degrees ' of + 1.3 kJ.mol-1. The pH and temperature dependence of k1*, of k2* and of K* were determined. From the second order rate constant k1* it was calculated that under physiological conditions the rate of spontaneous carbamate formation is of the same order as the maximal rate of methane formation and as the rate of spontaneous CO2 formation from HCO3- in methanogenic archaea, the latter being important as CO2 is mainly present as HCO3- which has to be converted to CO2 before it can react with MFR. An enzyme catalyzed carbamate formation thus appears not to be required for methanogenesis from CO2. Consistent with this conclusion is our finding that the rate of carbamate formation was not enhanced by cell extracts of Methanosarcina barkeri and Methanobacterium thermoautotrophicum or by purified formylmethanofuran dehydrogenase which catalyzes the reduction of N-carboxymethanofuran to N-formylmethanofuran. From the concentrations of 'CO2' and of 'MFR' determined by 1D-NMR spectroscopy and the pKa of H2CO3 and of MFRH+ the concentrations of CO2 and of MFR were obtained, allowing to calculate k1 (v1 = k1 [MFR] [CO2]). The second order rate constant k1 was found to be approximately 1000 M-1 x s-1 at 300 K and pH values between 7.0 and 8. 0 which is in the order of k1 values determined for other carbamate forming reactions by stopped flow. PMID- 10824098 TI - The 28.3 kDa FK506 binding protein from a thermophilic archaeum, Methanobacterium thermoautotrophicum, protects the denaturation of proteins in vitro. AB - Two families of FK506 binding protein (FKBP) type peptidyl-prolyl cis-trans isomerase (PPIase) have been found in Archaea. One is the 16-18 kDa short type FKBP family, and another is the 26-30 kDa long type FKBP family. The latter has a longer C-terminal region than the former. In this study, the 28.3 kDa long type FKBP gene from a thermophilic archaeum, Methanobacterium thermoautotrophicum, was expressed in Escherichia coli, and its gene product (MbFK) was characterized. The PPIase activity of MbFK was much lower than those of other FKBPs reported against oligopeptidyl substrates. MbFK protected green fluorescent protein (GFP) and rhodanese from thermal denaturation. Furthermore, MbFK suppressed the aggregation of chemically unfolded rhodanese and elevated the yield of its refolding although this activity was weaker than that of GroEL/ES. We made two deletion mutants, MbFK-N which lacked the C-terminal region, and MbFK-C which had only the C terminal region. Far-UV CD spectra of these mutants showed that their secondary structures did not change from that of the wild-type. Whereas the PPIase activity of MbFK-N was low but detectable, that of MbFK-C was undetectable. The MbFK-C protected the thermal protein aggregation, and possessed a weak but significant aggregation suppressing activity against chemically unfolded protein. However, the MbFK-N did not suppress the aggregation of chemically unfolded rhodanese while it protected heat induced aggregation of rhodanese. These results may indicate that aggregation suppressing activity of MbFK-W against chemically unfolded protein are exerted mainly by its C-terminal domain while both domains contribute to thermal protein aggregation suppression. PMID- 10824099 TI - Perlecan domain V of Drosophila melanogaster. Sequence, recombinant analysis and tissue expression. AB - The C-terminal domain V of the basement membrane proteoglycan perlecan was previously shown to play a major role in extracellular matrix and cell interactions. A homologous sequence of 708 amino-acid residues from Drosophila has now been shown to be 33% identical to mouse perlecan domain V. It consists of three laminin G-type (LG) and epidermal growth factor-like (EG) modules but lacks the EG3 module and a link region found in mammalian perlecans. Recombinant production of Drosophila perlecan domain V in mammalian cells yielded a 100-kDa protein which was folded into a linear array of three globular LG domains. Unlike the mouse counterpart, domain V from Drosophila was not modified by glycosaminoglycans and endogenous proteolysis, due to the absence of the link region. It showed moderate affinities for heparin and sulfatides but did not bind to chick alpha-dystroglycan or to various mammalian basement membrane proteins. A single RGD sequence in LG3 of Drosophila domain V was also incapable of mediating cell adhesion. Production of a proteoglycan form of perlecan (approximately 450 kDa) in one Drosophila cell line could be demonstrated by immunoblotting with antibodies against Drosophila domain V. A strong expression was also found by in situ hybridization and immunohistology at various stages of embryonic development and expression was localized to several basement membrane zones. This indicates, as for mammalian species, a distinct role of perlecan during Drosophila development. PMID- 10824100 TI - Structure of the sialic acid-containing O-specific polysaccharide from Salmonella enterica serovar Toucra O48 lipopolysaccharide. AB - Lipopolysaccharide was extracted from cells of Salmonella enterica serovar Toucra O48 and, after mild acid hydrolysis (1% AcOH, 1 h, 100 degrees C or 0.1 M NaOH AcOH, pH 4.5, 5 h, 100 degrees C), the O-specific polysaccharide was isolated and characterized. The core and an oligosaccharide containing a fragment of the repeating unit linked to the core region were also obtained, depending on hydrolysis conditions. On the basis of sugar and methylation analyses and NMR spectroscopy of the hydrolysis products, the biological repeating unit of the O specific polysaccharide was shown to be the following trisaccharide: -->4)-alpha Neup5Ac(2-->3)-L-alpha-FucpNAc(1-->3)-D-beta-Glc pNAc(1--> The polysaccharide O chain was substituted with a single molar equivalent of O-acetyl group, distributed between the Neu5Ac O-9 and O-7 positions, in an approximate ratio of 7 : 3. PMID- 10824101 TI - Biogenesis of the yeast frataxin homolog Yfh1p. Tim44-dependent transfer to mtHsp70 facilitates folding of newly imported proteins in mitochondria. AB - Tim44 is an essential component of the mitochondrial inner membrane protein import machinery. In this study we asked if Tim44 is of relevance in intramitochondrial protein folding. We investigated the role of Tim44 in the biogenesis of the authentic mitochondrial protein Yfh1p, the yeast homolog of mammalian frataxin, which was recently implicated in Friedreich ataxia. After inactivation of Tim44, binding of mitochondrial heat shock protein (mtHsp)70 to translocating Yfh1p and subsequent folding to the native state was nearly completely blocked. Residual amounts of imported Yfh1p showed an increased tendency to aggregate. To further characterize the functions of Tim44 in the matrix, we imported dihydrofolate reductase (DHFR) as a model protein. Depletion of Tim44 allowed import of DHFR, although folding of the newly imported DHFR was delayed. Moreover, the depletion of Tim44 caused a strongly reduced binding of mtHsp70 and Mge1 to the translocating polypeptide. Subsequent dissociation of mtHsp70 from imported DHFR was delayed, indicating that mtHsp70-substrate complexes formed independently of Tim44 differ from the complexes that form under the control of Tim44. We conclude that Tim44 not only plays a role in protein translocation but also in the pathways of mitochondrial protein folding. PMID- 10824102 TI - A novel protein kinase from Brassica juncea stimulated by a protozoan calcium binding protein. Purification and partial characterization. AB - A novel protein kinase (BjCCaBPk) from etiolated Brassica juncea seedlings has been purified and partially characterized. The purified enzyme migrated on SDS/PAGE as a single band with an apparent molecular mass of 43 kDa. The optimum pH for the kinase activity was 8.0. It was stimulated more than sixfold by the protozoa Entamoeba histolytica calcium binding protein EhCaBP (10.5 nM) but not by calmodulin (CaM) when used at equimolar concentration. Moreover the kinase also did not bind CaM-Sepharose. There was neither inhibition of the kinase activity in the presence of W-7 (a CaM antagonist), KN-62 (a specific calcium/CaM kinase inhibitor) and anti-CaM Ig, nor any effect on BjCCaBPk activity of staurosporine (a protein kinase C inhibitor). Furthermore a CaM-kinase specific substrate, syntide-2, proved to be a poor substrate for the BjCCaBPk compared with histone III-S. The phosphorylation of histone III-S involved serine residues. Southern and Northern blot analysis showed the presence of EhCaBP homologues in Brassica. The data suggest that BjCCaBPk may be a novel protein kinase with an affinity towards a calcium binding protein like EhCaBP. PMID- 10824103 TI - Mast cell tryptase from pig lungs triggers infection by pneumotropic Sendai and influenza A viruses. Purification and characterization. AB - A novel trypsin-type serine proteinase, which processes the precursors of the envelope fusion glycoproteins of pneumotropic Sendai and human influenza A viruses, was purified to homogeneity from pig lungs. On SDS/PAGE, the purified enzyme gave a protein band corresponding to about 32 kDa, and has an apparent molecular mass of 120 kDa, as determined by gel permeation chromatography. Immunohistochemical staining with antibodies against this enzyme revealed that the enzyme is located in pig lung mast cells. The N-terminal 44-amino-acid sequence of the enzyme exhibits about 80% identity with those of mast cell tryptases from other species. Of the inhibitors tested, di-isopropyl fluorophosphate, antipain, leupeptin, benzamidine and a few proteinaceous inhibitors, such as mucus protease inhibitor and aprotinin, inhibited this enzyme activity. Heparin stabilized the enzyme, but high-ionic-strength conditions did not, unlike for human mast cell tryptase. The purified enzyme efficiently processed the fusion glycoprotein precursor of Sendai virus and slowly processed hemagglutinin of human influenza A virus, and triggered the infectivity of Sendai virus in a dose-dependent manner, although human mast cell tryptase beta and rat mast cell tryptase (rat MCP-7) from lungs did not process these fusion glycoproteins at all. These results suggest that mast cell tryptase in pig lungs is the possible trigger of the pneumotropic virus infections. PMID- 10824104 TI - Molecular characterization of a puromycin-insensitive leucyl-specific aminopeptidase, PILS-AP. AB - The family M1 of Zn-dependent aminopeptidases comprises members of closely related enzymes which are known to be involved in a variety of physiologically important processes. On the basis of two highly conserved peptide motifs, we have identified a new member of this family by PCR amplification and cDNA-library screening. The longest ORF encodes a protein of 930 residues. It contains the HEXXH(X)18E Zn-binding motif and displays high homology to the other M1 family members except for its N-terminus for which a signal sequence of 20 residues can be predicted. This interpretation was supported by expressing fusion proteins formed with green fluorescent protein which localized to intracellular vesicles in COS-7 and BHK cells. Northern-blot analysis revealed ubiquitous expression of a major 3. 1-kb transcript. For enzymatic studies, the complete protein was expressed in Sf 9 insect cells. When aminoacyl beta-naphthylamides were used as substrates, efficient hydrolysis was only observed for Leu (and to a lesser extent Met). The activity was inhibited by chelators of bivalent cations and by other known aminopeptidase inhibitors, but surprisingly puromycin was without effect. This newly identified puromycin-insensitive leucyl-specific aminopeptidase is a signal-sequence-bearing member of family M1 and may be another example of the small subset of substrate-specific peptidases. PMID- 10824105 TI - Structural and functional divergence of a nuclear receptor of the RXR family from the trematode parasite Schistosoma mansoni. AB - We describe the cloning and functional characterization of Schistosoma mansoni retinoid-X-receptor (SmRXR; NR2B4-B), a novel member of the nuclear receptor superfamily from S. mansoni, a homologue of vertebrate retinoid-X-receptor. The DNA-binding C domain of SmRXR shows 80% sequence identity to both human RXRalpha and Drosophila ultraspiracle (USP), but a much lower level of conservation of the ligand-binding E domain (22-25% identity). Phylogenetic analysis places SmRXR within the RXR group as an early offshoot of this clade. SmRXR mRNA is expressed at all life-cycle stages but at higher levels in the free-living larval stages. However, the SmRXR protein is expressed at markedly different levels, being almost absent from eggs while present at the highest concentration in schistosomula. Recombinant SmRXR fails to bind to the consensus direct repeat response elements, either alone, or as a heterodimer with mouse retinoic acid receptor alpha or the Drosophila ecdysone receptor. However, the use of chimaeric constructions shows that the C domain of SmRXR will bind to conventional response elements as a heterodimer, and that its specificity is modified by the presence of the D and E domains. In accordance with these results, native SmRXR failed to transactivate the transcription of a reporter gene after cotransfection of mammalian cell lines. PMID- 10824106 TI - Annexin I is a stress protein induced by heat, oxidative stress and a sulfhydryl reactive agent. AB - Annexin I (also called lipocortin 1) is a 37-kDa member of the annexin family of proteins. It has been proposed to be involved in the regulation of cell growth and differentiation, apoptosis, and inflammation. Previously, we have reported that annexin I displays a chaperone-like function (Kim, G.Y., Lee, H.B., Lee, S.O., Rhee, H.J. & Na, D.S. (1997) Biochem. Mol. Biol. Int. 43, 521-528). To determine the possibility that annexin I is a stress protein, we examined whether expression of annexin I and annexin I mRNA increases in response to stresses in A549 and HeLa cells. Treatments of cells with heat, hydrogen peroxide or sodium arsenite resulted in (a) an increase in annexin I and annexin I mRNA and (b) translocation of annexin I from the cytoplasm to the nucleus and perinuclear region. The annexin I gene promoter region, cloned upstream of a reporter gene, was inducible in response to heat, hydrogen peroxide, and sodium arsenite. These results indicate that annexin I serves as a stress protein and annexins may constitute a new class of stress proteins. PMID- 10824107 TI - Regulation of secretory protein gene expression in paramecium role of the cortical exocytotic sites. AB - In cells that possess a regulated secretory pathway, exocytosis can lead to transcriptional activation of genes encoding products stored in secretory granules as well as genes required for granule biogenesis. With the objective of understanding this response, we have examined the expression of Paramecium secretory protein genes in different physiological and genetic contexts. The genes belong to the trichocyst matrix protein (TMP) multigene family, encoding polypeptides that form the crystalline matrix of the secretory granules, known as trichocysts. Approximately 1000 trichocysts per cell are docked at pre-formed cortical exocytotic sites. Their rapid and synchronous exocytosis can be triggered by vital secretagogues such as aminoethyldextran without harming the cells. Using this exocytotic trigger, we found that the transcription of TMP genes undergoes rapid, transient and co-ordinate 10-fold activation in response to massive exocytosis, leading to a 2.5-fold increase in the pool of TMP mRNA. Experiments with exocytosis-deficient mutants show that the secretagogue-induced increase in intracellular free calcium implicated in stimulus/secretion coupling is not sufficient to activate TMP gene expression. We present evidence that the state of occupation of the cortical exocytotic sites can affect TMP gene expression and suggest that these sites play a role in gene activation in response to exocytosis. PMID- 10824108 TI - Effects of acid pH and urea on the spectral properties of the LHII antenna complex from the photosynthetic bacterium Ectothiorhodospira sp. AB - The aim of this study was to investigate the spectral modifications of the LHII antenna complex from the purple bacterium Ectothiorhodospira sp. upon acid pH titration both in the presence and absence of urea. A blue shift specifically and reversibly affected the B850 band around pH 5.5-6.0 suggesting that a histidine residue most probably participated in the in vivo absorption red shifting mechanism. This transition was observed in the presence and absence of urea. Under strong chaotropic conditions, a second transition occurred around pH 2.0, affecting the B800 band irreversibly and the B850 reversibly. Under these conditions a blue shift from 856 to 842 nm occurred and a new and strong circular dichroism signal from the new 842 nm band was observed. Reverting to the original experimental conditions induced a red shift of the B850 band up to 856 nm but the circular dichroism signal remained mostly unaffected. Under the same experimental conditions, i.e. pH 2.1 in the presence of urea, part of the B800 band was irreversibly destroyed with concomitant appearance of a band around 770 nm due to monomeric bacteriochlorophyll from the disrupted B800. Furthermore, Gaussian deconvolution and second derivative of the reverted spectra at pH 8.0 after strong-acid treatment indicated that the new B850 band was actually composed of two bands centered at 843 and 858 nm. We ascribed the 858 nm band to bacteriochlorophylls that underwent reversible spectral shift and the 843 nm band to oligomeric bacteriopheophytin formed from a part of the B850 bacteriochlorophyll. This new oligomer would be responsible for the observed strong and mostly conservative circular dichroism signal. The presence of bacteriopheophytin in the reverted samples was definitively demonstrated by HPLC pigment analysis. The pheophytinization process progressed as the pH decreased below 2.1, and at a certain point (i.e. pH 1.5) all bacteriochlorophylls, including those from the B800 band, became converted to oligomeric bacteriopheophytin, as shown by the presence of a single absorption band around 843 nm and by the appearance of a single main elution peak in the HPLC chromatogram which corresponded to bacteriopheophytin. PMID- 10824109 TI - Conversion of pregnenolone to DHEA by human 17alpha-hydroxylase/17, 20-lyase (P450c17). Evidence that DHEA is produced from the released intermediate, 17alpha hydroxypregnenolone. AB - Most previous studies using reconstituted systems and fast kinetics suggest that the conversion of pregnenolone to dehydroepiandrosterone (DHEA; the precursor of androgen and estrogen biosynthesis) by P450c17 does not require the release of the intermediate 17alpha-OHPreg (a precursor of cortisol biosynthesis). With such a mechanism, it is difficult to conceive how high amounts of DHEA may be produced in some cells or tissues, such as the testis and cells from the adrenal reticularis, while in other tissues such as the fasciculata zone, high levels of 17alpha-OHPreg are synthesized. In this report, we address this matter using intact transfected cells, which better reflect the actual cellular conditions. Furthermore, by using transfected cells, we can conveniently analyze human enzymes, as we are not restricted by the availability of human tissues as in the case of methods using purified or partially purified enzymes. Using intact HEK 293 cells transfected with human P450c17 in culture, we showed, in a time course study of the transformation of pregnenolone, that there is an accumulation of 17alpha-OHPreg, and that, subsequently, the accumulated 17alpha-OHPreg decreases with a concomitant increase in DHEA production. The DHEA/17alpha-OHPreg ratio changes from 0.1 :1 after 1 h incubation to 50 : 1 after 20 h. This result strongly suggests that the transformation of Preg to DHEA proceeds through two steps in which DHEA is produced from the released intermediate 17alpha-OHPreg. We also show that high levels of substrate vs. enzyme concentration will lead to high hydroxylase activity whereas the reverse will increase the lyase activity. The result is in good agreement with recent observations suggesting that surrounding enzymes and steroids could modulate the lyase activity. Cotransfection of vectors expressing cytochrome b5 and NADPH cytochrome P450 reductase indicates that both are required for an optimum production of DHEA. PMID- 10824110 TI - Identification of a novel nuclear factor-kappaB sequence involved in expression of urokinase-type plasminogen activator receptor. AB - We have previously defined the promoter of human urokinase-type plasminogen activator receptor (uPAR) gene in a 188-bp fragment between bases -141 and +47 relative to the translation start site. Here, we report that a novel nuclear factor-kappaB (NF-kappaB)-like sequence (5'-GGGAGGAGTC-3') at -45 is located in the uPAR promoter and one of the two DNase I-protected regions, region I between bases -51 and -30. This NF-kappaB-like motif differs at positions 7-9 from the decameric consensus sequences of NF-kappaB (5'-GGGRNNYYCC-3' where R indicates A or G, Y indicates C or T, and N indicates any nucleotide) and at positions 1 and 7-9 from the kappaB-like motifs (5'-HGGARNYYCC-3' where H indicates A, C, or T, R indicates A or G, Y indicates C or T, and N indicates any nucleotide). Nuclear extracts from HCT116 cells contain proteins that specifically bind to the NF kappaB-like site at position -45. Mutation of the NF-kappaB-like motif decreased the binding of transcription factor NF-kappaB and reduced the uPAR promoter activity in comparison with the wild-type sequences. Co-transfection with a dominant negative I-kappaB kinase-2 expression vector reduced uPAR promoter activity by 65-75%. These results demonstrate that a previously uncharacterized NF-kappaB motif is required for uPAR promoter activity. PMID- 10824111 TI - Biochemical characterization of MI-ENG1, a family 5 endoglucanase secreted by the root-knot nematode Meloidogyne incognita. AB - A beta-1,4-endoglucanase named MI-ENG1, homologous to the family 5 glycoside hydrolases, was previously isolated from the plant parasitic root-knot nematode Meloidogyne incognita. We describe here the detection of the enzyme in the nematode homogenate and secretion and its complete biochemical characterization. This study is the first comparison of the enzymatic properties of an animal glycoside hydrolase with plant and microbial enzymes. MI-ENG1 shares many enzymatic properties with known endoglucanases from plants, free-living or rumen associated microorganisms and phytopathogens. In spite of the presence of a cellulose-binding domain at the C-terminus, the ability of MI-ENG1 to bind cellulose could not be demonstrated, whatever the experimental conditions used. The biochemical characterization of the enzyme is a first step towards the understanding of the molecular events taking place during the plant-nematode interaction. PMID- 10824112 TI - The oxidation and reduction reactions of bovine serum amine oxidase. A kinetic study. AB - The presteady-state and steady-state kinetics of bovine serum amine oxidase (BSAO) were analyzed by stopped-flow transient spectroscopy. A simplified model of the catalytic cycle was found to describe the experimental data and the rate constants of the individual steps were used to calculate Michaelis parameters that agree with the direct determinations. In spite of many studies on selected reactions from the catalytic cycle, this is amongst the first efforts to provide a comprehensive kinetic description of the reactions of BSAO, whose results can be compared with the steady-state parameters. The reoxidation reaction by dioxygen is more complex than previously thought, in agreement with a recent report [Su, Q. & Klinman, J.P. (1998) Biochemistry 37, 12513-12525], and occurs in at least two steps whose rate constants, previously undetermined, have been measured. The reaction of the oxidized enzyme with the amine substrate is poorly determined in this type of experiment, thus irreversible combination with aromatic hydrazine inhibitors was used as a model system, demonstrating that the mechanism and rate constants of their reaction is fully compatible with an accurate description of the catalytic cycle with the physiological substrate. These results constitute a simplified, yet complete and consistent, description of the catalytic cycle and offer an interesting comparison with those obtained on plant amine oxidases; two steps of the catalytic cycle are significantly slower in BSAO than in pea seedling or lentil seedling amine oxidases, namely the reoxidation and the trans-iminative proton abstraction occurring in the enzyme substrate complex. The former difference is rationalized as being due to the low to zero concentration of the semiquinolamine-radical intermediate, while the latter is less easily interpreted. PMID- 10824113 TI - Prolyl isomerases in a minimal cell. Catalysis of protein folding by trigger factor from Mycoplasma genitalium. AB - Peptidyl-prolyl cis/trans isomerases (PPIases) catalyze the isomerization of prolyl peptide bonds. Distinct families of this class of enzymes are involved in protein folding in vitro, whereas their significance in free living organisms is not known. Previously, we inspected the smallest known genome of a self replicating organism and found that Mycoplasma genitalium is devoid of all known PPIases except the trigger factor. Despite the extensive sequence information becoming available, most genes remain hypothetical and enzyme activities in many species have not been assigned to an open reading frame. Therefore, we studied the PPIase activity in crude extracts of M. genitalium. We showed that this is solely attributed to a single enzyme activity, the trigger factor. Characterization of this enzyme revealed that its PPIase activity resides in a central 12-kDa domain. Only the complete trigger factor is able to cis/trans isomerize extended peptide substrates, while the PPIase domain alone can not. The N- and the C-terminal domains of the trigger factor seem to function in binding of proteins as substrates, as demonstrated by protein refolding experiments, in which the complete trigger factor catalyzed protein refolding towards a model protein 500-fold more efficiently than the isolated central PPIase domain. Protein modeling studies suggest that the PPIase domain can fold in a similar way as the PPIase domain of FK506 binding proteins (FKBPs), one class of PPIases, despite only very limited sequence homology. Differences at the active site explain why this enzyme is not inhibited by FK506 in contrast with FKBPs. Trigger factor expressed in Escherichia coli confirms its additional chaperone functions, as shown by its association with chaperones GroEL and GroES after induction of misfolding. In contrast, the isolated PPIase-domain lacks any association with chaperones from E. coli. In summary, trigger factor of M. genitalium is the single folding isomerase of this organism, which harbors an enzymatically active PPIase domain with structural homology to FKBPs. Its additional domains confer its ability to be an efficient catalyst of protein folding. The protein folding machinery is conserved and shows a dual function as a chaperone and a prolyl isomerase. PMID- 10824114 TI - Interactions between the soluble domain I of nicotinamide nucleotide transhydrogenase from Rhodospirillum rubrum and transhydrogenase from Escherichia coli. Effects on catalytic and H+-pumping activities. AB - Nicotinamide nucleotide transhydrogenase from Escherichia coli is composed of two subunits, the alpha and the beta subunits, each of which contains a hydrophilic domain, domain I and III, respectively, as well as several transmembrane helices, collectively denoted domain II. The interactions between domain I from Rhodospirillum rubrum (rrI) and the intact or the protease-treated enzyme from E. coli was investigated using the separately expressed and purified domain I from R. rubrum, and His-tagged intact and trypsin-treated E. coli transhydrogenase. Despite harsh treatments with, e.g. detergents and denaturing agents, the alpha and beta subunits remained tightly associated. A monoclonal antibody directed towards the alpha subunit was strongly inhibitory, an effect that was relieved by added rrI. In addition, rrI also reactivated the trypsin-digested E. coli enzyme in which domain I had been partly removed. This suggests that the hydrophilic domains I and III are not in permanent contact but are mobile during catalysis while being anchored to domain II. Replacement of domain I of intact, as well as trypsin-digested, E. coli transhydrogenase with rrI resulted in a markedly different pH dependence of the cyclic reduction of 3-acetyl-pyridine-NAD+ by NADH in the presence of NADP(H), suggesting that the protonation of one or more protonable groups in domain I is controlling this reaction. The reverse reaction and proton pumping showed a less pronounced change in pH dependence, demonstrating the regulatory role of domain II in these reactions. PMID- 10824115 TI - Membranolytic selectivity of cystine-stabilized cyclic protegrins. AB - To correlate conformational rigidity with membranolytic selectivity of antimicrobial activity and cytotoxicity, we prepared six cyclic analogs of protegrin-1 (PG-1), an 18-residue cationic peptide with a broad-spectrum antimicrobial activity. These cyclic protegrins bear end-to-end peptide bonds together with varying numbers (zero to three) of cross-strand disulfide constraints. The most constrained analog is a cyclic tricystine protegrin (ccPG 3) containing three evenly spaced, parallel disulfide bonds. Antimicrobial assays against 10 organisms in low- and high-salt conditions showed that these cyclic protegrins were broadly active with different antimicrobial profiles against Gram positive and Gram-negative bacteria, fungi and one tested virus, HIV-1. Compared to PG-1, the cyclic tricystine ccPG 3 displayed approximately a 10-fold decrease in hemolytic activity against human cells and 6- to 30-fold improvement of membranolytic selectivity against six of the 10 tested organisms. In contrast, [DeltaSS]cPG 8, a cyclic protegrin with no disulfide bond, and [DeltaCys6,15]cPG 5, a cyclic mimic of PG-1 with one disulfide bond, exhibited activity spectra, potency, and cytotoxicity similar to PG-1. Circular dichroism showed that cyclic protegrins containing with one to three cystine bonds displayed some degree of beta-strand structures in water/trifluoroethanol or phosphate-buffered solutions. Collectively, our results indicate that cyclic structures are useful in the design of antimicrobial peptides and that an increase in the conformational rigidity of protegrins may confer membranolytic selectivity that dissociates antimicrobial activity from hemolytic activity. PMID- 10824116 TI - Molecular cloning and characterization of murine and human N-acetylglucosamine kinase. AB - N-Acetylglucosamine is produced by the endogenous degradation of glycoconjugates and by the degradation of dietary glycoconjugates by glycosidases. It enters the pathways of aminosugar metabolism by the action of N-acetylglucosamine kinase. In this study we report the isolation and characterization of a cDNA clone encoding the murine enzyme. An open reading frame of 1029 base pairs encodes 343 amino acids with a predicted molecular mass of 37.3 kDa. The deduced amino-acid sequence contains matches of the sequences of eight peptides derived from tryptic cleavage of rat N-acetylglucosamine kinase. The recombinant murine enzyme was functionally expressed in Escherichia coli BL21 cells, where it displays N acetylglucosamine kinase activity as well as N-acetylmannosamine kinase activity. The complete cDNA sequence of human N-acetylglucosamine kinase was derived from the nucleotide sequences of several expressed sequence tags. An open reading frame of 1032 base pairs encodes 344 amino acids and a protein with a predicted molecular mass of 37.4 kDa. Similarities between human and murine N acetylglucosamine kinase were 86.6% on the nucleotide level and 91.6% on the amino-acid level. Amino-acid sequences of murine and human N-acetylglucosamine kinase show sequence similarities to other sugar kinases, and all five sequence motifs necessary for the binding of ATP by sugar kinases are present. Tissue distribution of murine N-acetylglucosamine kinase revealed an ubiquitous occurrence of the enzyme and a very high expression in testis. The size of the murine mRNA was 1.35 kb in all tissues investigated, with the exception of testis, where it was 1.45 kb mRNA of the murine enzyme was continuously expressed during mouse development. mRNA of the human enzyme was expressed in all investigated human tissues, as well as in cancer cell lines. In both the tissues and the cancer cell lines, the human mRNA was 1.35 kb in size. PMID- 10824117 TI - Studies on the recycling of the transferrin receptor in Trypanosoma brucei using an inducible gene expression system. AB - Uptake of host transferrin in bloodstream forms of Trypanosoma brucei is mediated by a heterodimeric, glycosylphosphatidylinositol-anchored receptor. After endocytosis, transferrin is delivered to lysosomes where it is proteolytically degraded. Whether the heterodimeric transferrin receptor is returned to mediate several cycles in ligand uptake is undefined. By using an inducible gene expression system we provide evidence for recycling of the transferrin receptor in bloodstream forms of T. brucei. The metabolic half-life of the transferrin receptor in bloodstream-form trypanosomes is determined to be 7 h which is comparable to the half-lives of recycling receptors in mammalian cells. The cycling time of the trypanosomal transferrin receptor is calculated to be 11 min. By means of the half-life and the cycling time, we calculated that each receptor is recycled 60 times before being degraded on average. PMID- 10824118 TI - Structure and chromosomal localization of human and mouse genes for hematopoietic prostaglandin D synthase. Conservation of the ancestral genomic structure of sigma-class glutathione S-transferase. AB - Hematopoietic prostaglandin D synthase (H-PGDS) is the key enzyme for the production of the D and J series of prostanoids, and the first recognized vertebrate homolog of sigma-class glutathione S-transferase (GST). We isolated the genes and cDNAs for human and mouse H-PGDSs. The human and mouse cDNAs contained a coding region corresponding to 199 amino-acid residues with calculated molecular masses of 23 343 and 23 226, respectively. Both H-PGDS proteins recombinantly expressed in Escherichia coli showed bifunctional activities for PGDS and GST, and had almost the same catalytic properties as the rat enzyme. Northern analyses demonstrated that the H-PGDS genes were expressed in a highly species-specific manner. Whereas the human gene was widely distributed, in contrast, the mouse gene was detected only in samples from oviduct and skin. By fluorescence in situ hybridization, the chromosomal localization of the human and mouse H-PGDS genes were mapped to 4q21-22 and 3D-E, respectively. The human and mouse H-PGDS genes spanned approximately 41 and 28 kb, respectively, and consisted of six exons divided by five introns. The exon/intron boundaries of both genes were completely identical to those of the sigma-class GST subfamily, although the amino-acid sequences of the latter were only 17.0-21.5% identical to those of either H-PGDS. These findings suggest that the H-PGDS genes evolved from the same ancestral gene as the members of the sigma class GST family. PMID- 10824120 TI - Arginine-based structures are specific inhibitors of cathepsin C. Application of peptide combinatorial libraries. AB - Novel synthetic peptide inhibitors of lysosomal cysteine proteinase cathepsin C have been designed through the use of soluble peptide combinatorial libraries. The uncovered structural inhibitory module consists of the N-terminal cluster of L-arginine residues. Its modification with D-amino acids or arginine derivatives did not increase the inhibition strength. Inhibitory potency of oligoarginines improves with the elongation of peptide chain reaching a maximum for octa-L arginine. The oligoarginines specifically interact with the cathepsin C active site as shown by competitive-type inhibition kinetics (Ki approximately 10-5 M) and intrinsic fluorescence measurements. The inhibitory interaction of oligoarginines is established through the specific spatial contact of a net of guanidino groups in the arginine side-chains, as indicated by comparison with inhibitory action of low molecular mass guanidine derivatives (Ki approximately 10-3 M). Nonarginine polyionic compounds cannot mimic the inhibitory effect of oligoarginines. The arginine-based peptide inhibitors were selective towards cathepsin C among other cysteine proteinases tested. PMID- 10824119 TI - Cloning of MMP-26. A novel matrilysin-like proteinase. AB - A cDNA encoding a novel human matrix metalloproteinase (MMP), named MMP-26, was cloned from fetal cDNA. The deduced 261-amino-acid sequence is homologous to macrophage metalloelastase (51.8% identity). It includes only the minimal characteristic features of the MMP family: a signal peptide, a prodomain and a catalytic domain. As with MMP-7, this new MMP does not comprise the hemopexin domain, which is believed to be involved in substrate recognition. A study of MMP 26 mRNA steady states levels reveals, among the tissue examined, a specific expression in placenta. MMP-26 mRNA could also be detected in several human cell lines such as HEK 293 kidney cells and HFB1 lymphoma cells. Recombinant MMP-26 was produced in mammalian cells and used to demonstrate a proteolytic activity of the enzyme on gelatin and beta-casein. PMID- 10824121 TI - Detection of modifications in the glucose metabolism induced by genetic mutations in Saccharomyces cerevisiae by 13C- and H-NMR spectroscopy. AB - NMR spectroscopy may offer a suitable technique to characterize the glucose metabolism in response to genetic mutations in cells. The effects of various genetic modifications in Saccharomyces cerevisiae yeast were investigated using 13C- and 1H-NMR spectroscopy associated with biochemical techniques. Cells were incubated with [1-13C]glucose in order to study glucose consumption and the formation of various end-products (ethanol, trehalose, glycerol, glutamate and amino acids) as a function of time. Two types of genetic modifications were studied in S. cerevisiae. A genetic modification deleted the N-terminal part of the TFC7 protein which is the smallest subunit (tau55) of the TFIIIC transcription factor. One secondary effect of this mutation was a large deletion of mitochondrial DNA giving the rho-phenotype. The other genetic modification corresponded to the disruption of the HUF gene; the mutated cells were rho+ like the reference strain. Both mutations increase the glycolysis rate and glycerol synthesis and decrease trehalose production. The most modified cells, which contain both TFC7 deletion and HUF gene disruption, utilize glucose in the most extreme manner as in these cells the largest production of the two glycolytic products (ethanol and glycerol) and the smallest trehalose formation occur. The HUF gene disruption serves as a positive modulator of glycolysis and respiration. However, the TFC7 deletion, associated with the phenotype rho-, induces the most damage in the cellular function, dramatically altering the behaviour of the Krebs cycle. The cycle becomes blocked at the level of 2-oxoglutarate, detected by a characteristic pattern of the 13C-NMR glutamate spectra. These NMR spectra corroborate the phenotypic data, the rho-phenotype corresponding to deletions of mitochondria DNA which block all mitochondria protein synthesis and render the cells unable to derive energy from respiration. Moreover, as a consequence of the Krebs cycle blocking, alanine formation is also observed. PMID- 10824122 TI - Effects of volatile anesthetic on the Ca2+-ATPase activation by dimerization. Distance-dependent quenching analysis and fluorescence energy transfer studies. AB - The phenomenological distance-dependent quenching (DDQ) model was employed to investigate the character of the interaction between volatile anesthetics (VAs) and the plasma membrane Ca2+-ATPase (PMCA). The simultaneous analysis of the frequency-domain and steady-state data of tryptophan (Trp) fluorescence quenching by a VA points to a specific character of the apparent quenching effect of the VA, possibly arising from a significant contribution of static quenching. The apparent contributions of both static and dynamic quenching may be due to VA binding in the PMCA, which results in the modification of the conformational substates of the enzyme. To characterize further the molecular consequences of VA binding, we investigated its effects on the process of PMCA activation by self association. VA shifted the equilibrium from enzyme dimers to monomers, as monitored by the loss of fluorescence energy transfer. The shift was apparently due to the VA-induced decrease in the affinity of PMCA molecules for self association. Addition of a large molecular mass dextran to increase the proximity between enzyme monomers induced re-association of the VA-impaired PMCA, while the Ca2+-ATPase activity was not recovered. The results are congruent with a dual VA effect on PMCA, a shift in the monomer/dimer equilibrium, and an inactivation of both monomers and dimers. PMID- 10824123 TI - Genomic structure and chromosomal localization of the human hepatocyte growth factor activator inhibitor type 1 and 2 genes. AB - Hepatocyte growth factor activator inhibitor type 1 (HAI-1) and type 2 (HAI-2) are recently discovered Kunitz-type serine protease inhibitors which can be purified and cloned from human stomach cancer cell line MKN45 as specific inhibitors against hepatocyte growth factor activator (HGFA). HAI-2 was identical with the protein originally reported as placental bikunin. Both proteins contain two Kunitz inhibitor domains (KDs), of which the first domain (KD1) is mainly responsible for the inhibitory activity against HGFA, and are expressed ubiquitously in various tissues. In this study, we cloned the genes coding for these two structurally similar proteins by screening of human genomic bacterial artificial chromosome (BAC) library and their genomic structures were compared. HAI-1 and -2 genes consist of 11 and 8 exons spanning 12 kbp and 12.5 kbp, respectively. Three exons were inserted between KD1 and KD2 of each gene, of which the middle one was the low-density lipoprotein (LDL) receptor-like domain (HAI-1) and the testis specific exon (HAI-2). Apparently homologous regions between HAI-1 and -2 were not found in 5'-flanking region and neither TATA nor CAAT box was present. The genes were mapped to chromosome 15q15 (HAI-1) and 19q13.11 (HAI-2). These results suggested that although HAI-1 and -2 genes might be derived from same ancestor gene, they acquired distinctive in vivo roles during their evolution. PMID- 10824124 TI - Keratan sulfates from bovine tracheal cartilage structural studies of intact polymer chains using H and 13C NMR spectroscopy. AB - Intact keratan sulfate chains derived from bovine tracheal cartilage have been examined using both one-dimensional methods and the two-dimensional experiments COSY-45 and TOCSY for homonuclear shift correlations and a modified COLOC (correlated spectroscopy for long-range couplings) approach for 13C-1H shift correlations. Partial 1H and 13C NMR signal assignments for residues within the intact polymer chain are reported; data derived from the repeat region signals and from chain cap residues are assigned by comparison with published data derived from oligosaccharides obtained through cleavage of keratan sulfate polymer chains using keratanase and keratanase II and are discussed in detail. The one-dimensional spectra for both 1H and 13C nuclei contain highly crowded signal clusters for which data analysis is not directly possible. COSY-45 analysis allow the correlation and assignment of many proton resonances located within the 3.4-4.8 p.p.m. chemical shift region while from the C/H correlation spectrum data are assignable for some signals within the complex set of carbon resonances which fall in the region between 68 and 86 p.p.m., This work using material from tracheal cartilage has permitted the first detailed combined 1H and 13C NMR examination of the primary keratan sulfate polymer structure; this sequence forms the basis for the more complex members of the keratan sulfate family present in other tissues such as articular cartilage and cornea where further residues such as (alpha1-3)-linked fucose and (alpha2-6)-linked N acetylneuraminic acid are also present. This nondestructive method of analysis complements the currently available degradative methods for structure determination which may then subsequently be utilized. PMID- 10824125 TI - Physicochemical characteristics of triacyl lipid A partial structure OM-174 in relation to biological activity. AB - The triacylated lipid A partial structure OM-174 was characterized in detail using a variety of physical and biological techniques. OM-174 aggregates adopt the micellar HI structure. The temperature (Tc) of the gel to liquid-crystalline phase transition of the hydrocarbon chains is 0 degrees C, from which high fluidity of the acyl chains at 37 degrees C can be deduced. The molecular area of a single OM-174 molecule at a surface pressure of 30 mN x m-1 is 0.78 +/- 0.04 nm2. Conformational analyses, using IR spectroscopy, of the behavior of the various functional groups of OM-174 as compared with hexa-acyl lipid A suggest altered hydration of the phosphate charges and unusually strong hydration of the ester groups, which is probably related to the high accessibility of these groups to water in the micellar aggregate structure. OM-174 was shown to intercalate into a phospholipid membrane corresponding to the macrophage membrane within seconds in the presence, and within minutes to hours in the absence, of LPS binding protein. In the Limulus amebocyte lysate assay, the triacyl lipid A is more than 105-fold less active than hexa-acyl lipid A, but only 10-fold less active in inducing IL-6 in human mononuclear cells, and equally active in inducing NO production in murine macrophages. These findings are used to explain the mechanism of the lipid A-induced cell activation. PMID- 10824126 TI - Rapid formation and high diffusibility of actin-cofilin cofilaments at low pH. AB - Cofilin is a small actin-binding protein that is known to bind both F-actin and G actin, severing the former. The interaction of cofilin with actin is pH sensitive, F-actin being preferentially bound at low pH and G-actin at higher pH, within the physiological range. Diffusion coefficients of F-actin with cofilin were measured by the fluorescence recovery after photobleaching (FRAP) technique. This has the potential for simultaneous and direct measurement of average polymer length via the average diffusion coefficient of the polymers (DLM) as well as the fraction of polymerized actin, fLM, present in solution. In the range of cofilin actin ratios up to 1 : 1 and at both pH 6.5 and pH 8.0, the diffusion coefficients of the polymers increased with the amount of cofilin present in the complex, in a co-operative manner to a plateau. We interpret this as indicating co-operative binding/severing and that filaments less than a certain length cannot be severed further. Under the conditions used here, filaments were found to be more motile at pH 6.5 than at pH 8.0. At pH 8.0, some actin is expected to be sequestered as ADP-actin-cofilin complexes, with the remaining actin being present as long slowly diffusing filaments. At pH 6.5, however, cofilin binds to F-actin to form short rapidly diffusing cofilaments. These filaments form very rapidly from cofilin-actin monomeric complexes, possibly indicating that this complex is able to polymerize without dissociation. These findings may be relevant to the nuclear import of actin-cofilin complexes. PMID- 10824127 TI - Transient N-acetylgalactosaminylation of mannosyl phosphate side chain in Paramecium primaurelia glycosylphosphatidylinositols. AB - The surface antigens of the free-living protozoan Paramecium primaurelia belong to the family of glycosylphosphatidylinositol (GPtdIns)-anchored proteins. Using a cell-free system prepared from P. primaurelia, we have described the structure and biosynthetic pathway for GPtdIns glycolipids. The core glycans of the polar glycolipids are modified by a mannosyl phosphate side chain. The data suggest that the mannosyl phosphate side chain is added onto the core glycan in two steps. The first step involves the phosphorylation of the GPtdIns trimannosyl conserved core glycan via an ATP-dependent kinase, prior to the addition of the mannose linked to the phosphate group. We show that dolichol phosphate mannose is the donor of all mannose residues including the mannose linked to phosphate. Furthermore, we were able to identify in vitro a hydrophilic intermediate containing an additional N-acetylgalactosamine linked to the mannosyl phosphate side chain. The addition of this purified hydrophilic radiolabelled intermediate into the cell-free system leads to a loss of the GalNAc residue and its conversion to the penultimate intermediate having only mannosyl phosphate as a side chain. Together the data indicate that the GalNAc-containing intermediate is a transitional intermediate. We suggest that the GalNAc-containing intermediate is essential for biosynthesis and maturation of GPtdIns precursors. It is hypothesized that this oligosaccharide processing in the course of GPtdIns biosynthesis is required for the translocation of GPtdIns from the cytoplasmic side of the endoplasmic reticulum to the luminal side. PMID- 10824128 TI - Foam fractionation of binary mixtures of lysozyme and albumin. AB - A nitrogen gas-based foam fractionation method was employed to separate model proteins, bovine serum albumin (BSA) and hen egg white lysozyme, from each other. Fractionation was characterized by the separation ratio and by recovery of proteins in the retentate as a function of the nominal pore size of the gas dispersion frit and solution conditions (pH and ionic strength). For binary mixtures of the proteins at pH 7.4, and ionic strength (mu) of 0.18 M, the recovery of lysozyme and the separation ratio were both dependent on the frit size employed to generate the foam. At low ionic strength (mu = 0.01 M), separation was only somewhat greater with the small pore size frits, although at values significantly lower than those found for high ionic strength. The diminished separations appear to be due to the only slight changes in recoveries observed for BSA and lysozyme.%Separation ratios of lysozyme from BSA in solutions either of high or low ionic strength were maximal at pH values equal to or less than the isoelectric point (pI) of BSA. Separation ratios were lower when foaming was carried out under low compared with high ionic strength. The recovery of lysozyme was enhanced by foaming from solutions of low pH and high ionic strength. Recoveries of BSA were greatest when the molecule was negatively charged. Electrical interactions between the positively charged lysozyme and negatively charged BSA may explain the diminished separation ratios and enhanced recoveries. Enzyme activity studies of lysozyme remaining in the retentate showed no change from prefoam activity. PMID- 10824129 TI - The effect of ultrasonic vibration on the compaction characteristics of paracetamol. AB - An ultrasonic (US) compaction rig has been developed that is capable of providing compaction pressure together with high-power ultrasonic vibrations of 20 kHz to a powder or granular material in a die. The rig has been used to investigate the effect of US on the compaction properties of paracetamol, a drug that produces tablets that are weak and frequently exhibit capping. It was found that coherent paracetamol tablets could be prepared by US-assisted compaction at pressures as low as 20 to 30 MPa. Application of US before and after compaction was not found to be as effective as US applied during compaction. The breaking forces of the tablets produced with US applied during compaction were found to be consistently significantly higher than when compaction was performed conventionally or with US applied before or after compaction. The application of US during compaction made it possible to increase tablet breaking force, typically by a factor of 2 to 5. It was concluded that pressure should be applied together with US to achieve a better acoustical contact, which is required to transmit vibrations from the horn to the material and also to bond the surfaces of the particles. US application during compaction also resulted in an increase in apparent density, in relation to the apparent density of conventionally prepared paracetamol tablets, of up to 12.8%. US appears to improve particle rearrangement and provide energy for partial melting of particle asperities and subsequent fusion of particle surfaces, thus increasing interparticulate bonding. Development of solid bridges between the particles during US-assisted compaction was observed on scanning electron photomicrographs. Solid bridge formation was thought to result in a reduction of void space, which in turn reduced the rate of water penetration into the compacts and consequently increased tablet disintegration and drug dissolution times. It was found that the results of US-assisted compaction are influenced by formulation and US time. An increase in binder (polyvinylpyrrolidone) concentration and/or US time resulted in a significant increase in the breaking forces of paracetamol tablets produced with US. When paracetamol was mixed with a second material, such as dicalcium phosphate dihydrate and microcrystalline cellulose, stronger compacts were prepared by US assisted compaction compared with the tablets containing no filler. Positive interactions were considered to have occurred as a result of US-induced bonding between the two materials. Overall, the application of US was found to significantly improve the compaction properties of paracetamol. PMID- 10824130 TI - Total deposition of therapeutic particles during spontaneous and controlled inhalations. AB - Treatment of systemic diseases by means of the inhalation route is hampered by uncertainties of the drug dose applied by inhalation. In this study, the hypothesis was tested that by standardization of the breathing maneuver used for inhalation, the interindividual variability of the dose deposited intrathoracically can be reduced. Therefore, breathing pattern during routine inhalations with jet nebulizers was measured in 18 patients with lung disease. Using monodisperse 3 microm particles, total deposition was then assessed for the measured spontaneous and for three controlled, slow breathing patterns. Particle deposition for the three controlled breathing patterns was additionally measured in 14 healthy subjects. The study has shown that within the study population the inhaled air volume and flow rate were quite different. Consequently, total particle deposition varied between 20 and 95%, depending on breathing pattern. For controlled, slow breathing patterns, deposition was on average higher, intersubject variability of deposition was smaller, and differences in deposition between healthy subjects and patients were negligible. Therefore, to perform efficient systemic treatment with aerosolized drugs, controlled, slow breathing patterns should be used. PMID- 10824131 TI - Sustained suppression of pituitary-gonadal axis with an injectable, in situ forming implant of leuprolide acetate. AB - The objective of these studies was to develop a leuprolide acetate depot based on an in situ forming drug delivery system (Atrigel(R)) to suppress the pituitary gonadal axis and in turn the serum testosterone to chemical castration levels for a period of at least 3 months. Formulations with biodegradable lactide/glycolide copolymers that varied in molecular weight, lactide/glycolide ratio, and hydrophilicity were evaluated in rats for their efficacy by measuring serum testosterone levels. The effect of polymer irradiation was also investigated. Molecular weight of the polymers was characterized by gel-permeation chromatography, and retrieved implants at the termination of animal studies were assayed for residual drug content by high-performance liquid chromatography. These initial rat studies showed that a formulation containing a 75/25 lactide/glycolide copolymer dissolved in N-methyl-2-pyrrolidone with 3% w/w leuprolide acetate suppressed serum testosterone for a period of 3 months or longer. This formulation with its advantages of biodegradability, biocompatibility, ease of injection, and no need for removal after use should be beneficial in treating patients with hormonal-dependent prostate and mammary cancers, endometriosis, and precocious puberty. In addition, this formulation with its simple manufacturing process is expected to provide an economic benefit to the user compared with products currently available on the market. PMID- 10824132 TI - Effect of conformation on the rate of deamidation of vancomycin in aqueous solutions. AB - The instability of vancomycin, a glycopeptide antibiotic, limits its shelf-life because the deamidation of its asparagine residue results in the formation of a zwitterion with limited aqueous solubility. Analysis of the pH-rate profile for vancomycin indicates that the deamidation reaction is notably sensitive to the ionic state of the molecule. This observation results in a hypothesis in which the ionic state of vancomycin may influence the conformation of the molecule and therefore affect its reactivity. Two-dimensional nuclear magnetic resonance (NMR), homonuclear Hartmann-Hahn (HOHAHA) and rotating frame Overhauser enhancement spectroscopy (ROESY) information combined with molecular dynamic simulations were used to estimate the apparent conformation of vancomycin in aqueous solution at pH 4 and pH 9 where the molecule exists primarily as a monocation and monoanion, respectively. The apparent conformation for vancomycin at pH 4 is compact, and the proximity of the backbone amide nitrogen to the side chain carbonyl carbon of asparagine is favorable for the rapid formation of the cyclic imide intermediate, thus increasing its reactivity. The apparent conformation for vancomycin at pH 9, however, is expanded in comparison with the conformation at pH 4, and the increase in distance between the reacting atoms leads to slower cyclic imide formation and thus decreased intrinsic reactivity. That cyclic imide formation was rate limiting at both pH values was confirmed by cyclic imide isolation and stability estimation. It becomes apparent from the analysis of the pH-rate and conformational profiles of vancomycin that the deamidation rate of vancomycin is largely influenced by the ionization state of the N-methyl leucine nitrogen. PMID- 10824133 TI - Enantioselective binding of propranolol, disopyramide, and verapamil to human alpha(1)-acid glycoprotein. AB - We investigated the binding of propranolol (PL), disopyramide (DP), and verapamil (VP) enantiomers by human alpha(1)-acid glycoprotein (AGP; also called orosomucoid) and the relationships between the extent of drug binding and lipophilicity, desialylation, and genetic variants of AGP. Desialylation had little effect on the affinity of AGP for the drugs tested. The percentage binding correlated significantly with the partition coefficients for the drugs tested. Each enantiomer was competitively displaced from AGP by another enantiomer of the same drug, suggesting that they bind to the same site. However, the enantiomers bound to AGP with stereospecific affinities; the (-)-isomers of DP and VP had higher Kd values (4.27 and 4.97 microM, respectively) than the (+)-isomers (1.51 and 2.48 microM, respectively). When enantiomers of the different drugs were used in competitive binding experiments, VP binding was only partially inhibited by DP. This result suggested that drug binding is specific to different variants of AGP (A, F1, S). DP was found to specifically bind to variant A, whereas PL and VP bind to both A and F1/S variants. PMID- 10824134 TI - Degradation pathways of a peptide boronic acid derivative, 2-Pyz-(CO)-Phe-Leu B(OH)(2). AB - The peptide boronic acid derivative 2-Pyz-(CO)-Phe-Leu-B(OH)(2) is a potent inhibitor of 20S proteasome and a proposed anticancer agent. During preformulation studies, the compound presented erratic stability behavior. Efforts were made to isolate and identify the degradation products, thereby helping to identify possible mechanisms for the degradation. The reaction of 2 Pyz-(CO)-Phe-Leu-B(OH)(2) with hydrogen peroxide not only provided a convenient way to isolate the initial degradation products seen from hydrolysis in aqueous buffers but also showed that the major, initial degradation pathway was probably oxidative in nature. The isolated degradation products were characterized by nuclear magnetic resonance spectroscopy, mass spectrometry, and optical rotation dispersion. In the presence of hydrogen peroxide, the boronic acid group was cleaved from 2-Pyz-(CO)-Phe-Leu-B(OH)(2) to give an alcohol with an apparent retention of the original stereochemistry. Subsequent isomerization and further hydrolysis were then seen. Surprisingly, added ascorbate and EDTA accelerated rather than inhibited degradation. Degradation of 2-Pyz-(CO)-Phe-Leu-B(OH)(2) under acidic and basic conditions seemed to be mediated by an initial oxidative degradation pathway similar to that seen with the peroxide. PMID- 10824135 TI - Identification of a yellow impurity in aged samples of aqueous butamben suspension: evidence for the oxidative degradation of poly(ethylene glycol). AB - Butamben (butyl p-aminobenzoate) has been formulated to provide long-acting treatment for chronic pain. The suspension, which contains poly(ethylene glycol) and polysorbate 80, was found to yellow under ambient conditions if not adequately protected from oxygen. The impurity responsible for the color was isolated and identified on the basis of nuclear magnetic resonance spectroscopy and mass spectrometry. The compound is an oxalamidine, which is formally the condensation product of oxalic acid with four equivalents of butamben, and may be formed by the reaction of butamben with an oxidation product of poly(ethylene glycol). PMID- 10824136 TI - Mechanism(s) of in vitro percutaneous absorption enhancement of tamoxifen by enhancers. AB - The effects of enhancers (5% terpenes; i.e., eugenol, limonene, and menthone) in combination with 50% propylene glycol in water (50% PG) on the in vitro percutaneous absorption of tamoxifen through the porcine epidermis, on biophysical changes in the stratum corneum (SC) lipids, on macroscopic barrier properties, and on binding of the drug to the SC were investigated. These enhancers in combination with 50% PG significantly increased (p<0.05) the permeability coefficient of tamoxifen in comparison with that of the control (50% PG in water). Fourier transform infrared spectroscopy (FT-IR) was employed to investigate the biophysical changes in the SC lipids. The FT-IR results showed that treatment of the SC with 5% terpenes/50% PG did not shift the asymmetric and symmetric C-H stretching absorbances peak positions to higher wavenumbers but resulted in a decrease in the peak heights and areas in comparison with the untreated SC. Treatment with menthone and limonene in combination with 50% PG significantly increased (p<0.05) the partition coefficient of tamoxifen in comparison with treatment with 50% PG alone. Also, exposure of the SC to 5% terpenes in combination with 50% PG significantly increased (p < 0.05) the in vitro transepidermal water loss (TEWL) in comparison with 50% PG alone. Thus, an enhancement by menthone, eugenol, and limonene in the permeability of the SC to tamoxifen is due to lipid extraction and macroscopic barrier perturbation. Moreover, the effective diffusion coefficient of tamoxifen through the epidermis was enhanced following the treatment with either 5% eugenol/50%PG or 5% limonene/50%PG compared with 50%PG alone. PMID- 10824137 TI - Transport of valganciclovir, a ganciclovir prodrug, via peptide transporters PEPT1 and PEPT2. AB - In clinical trials, valganciclovir, the valyl ester of ganciclovir, has been shown to enhance the bioavailability of ganciclovir when taken orally by patients with cytomegalovirus infection. We investigated the role of the intestinal peptide transporter PEPT1 in this process by comparing the interaction of ganciclovir and valganciclovir with the transporter in different experimental systems. We also studied the interaction of these two compounds with the renal peptide transporter PEPT2. In cell culture model systems using Caco-2 cells for PEPT1 and SKPT cells for PEPT2, valganciclovir inhibited glycylsarcosine transport mediated by PEPT1 and PEPT2 with K(i) values (inhibition constant) of 1.68+/-0.30 and 0.043+/- 0.005 mM, respectively. The inhibition by valganciclovir was competitive in both cases. Ganciclovir did not interact with either transporter. Similar studies done with cloned PEPT1 and PEPT2 in heterologous expression systems yielded comparable results. The transport of valganciclovir via PEPT1 was investigated directly in PEPT1-expressing Xenopus laevis oocytes with an electrophysiological approach. Valganciclovir, but not ganciclovir, induced inward currents in PEPT1-expressing oocytes. These results demonstrate that the increased bioavailability of valganciclovir is related to its recognition as a substrate by the intestinal peptide transporter PEPT1. This prodrug is also recognized by the renal peptide transporter PEPT2 with high affinity. PMID- 10824138 TI - A time-dependent expression for thixotropic areas. Application to aerosil 200 hydrogels. AB - Experimental determination of the areas enclosed under the up curve, sigma = sigma(;gamma), corresponding to time zero, and the down curve, which would be obtained following complete rheodestruction of the system structure, is not an easy matter. In the present study a semi-empirical procedure for calculating the hypothetical values of these parameters is proposed. With these parameters it is possible to obtain a time-dependent expression for the thixotropic areas, S(T). This procedure has been applied to the study of the thixotropic behavior of Aerosil 200 hydrogels at different concentrations. The comparative analysis of these systems has been realized by studying the relative thixotropic areas and relative thixotropic rates that are obtained from S(T). PMID- 10824139 TI - Physicochemical and solubilization properties of N, N-dimethyl-N-(3 dodecylcarbonyloxypropyl)amineoxide: a biodegradable nonionic surfactant. AB - The physicochemical characteristics of N,N-dimethyl-N-(3-dodecylcar- bonyloxypropyl)amineoxide (DDCPNO), a biodegradable analogue of a N, N dimethylalkylamine-N-oxide, are compared with those of N, N-dimethyldodecylamine N-oxide (DDNO) to establish the effect on the properties of DDNO of inserting a propoxy linker between the hydrophobic tail and hydrophilic head region. Surface tension measurements gave a critical micelle concentration of 0.33 mM for DDCPNO compared with a value of 1.57 mM for DDNO, suggesting that the former was the more hydrophobic surfactant. This result was confirmed by laser light scattering studies in which total intensity light scattering indicated the presence of DDCPNO micelles of aggregation number 85.0. Photon correlation spectroscopy studies yielded a limiting hydrodynamic diameter of 4.0 nm in comparison with values of 57.5 and 3.3 nm obtained for the aggregation number and the limiting hydrodynamic size, respectively, of DDNO micelles. Studies demonstrated that neither a dilute aqueous solution of DDCPNO or DDNO exhibited a cloud point within the temperature range 293-373 K. Solubilization studies showed that the capacity of DDCPNO micelles for a range of drugs of varying size and polarity was less than that observed with DDNO micelles at an equivalent surfactant concentration. As a further measure of solubilization, the ability of DDCPNO to form oil-in-water microemulsions with a range of ethyl ester oils was investigated and found to be slightly higher than that achieved with DDNO. Together these studies suggest that the presence of the semipolar linker significantly alters the properties of this low molecular weight surfactant. PMID- 10824140 TI - Synthesis and characterization of surface-hydrophobic ion-exchange microspheres and the effect of coating on drug release rate. AB - Biodegradable, dextran-based ion-exchange microspheres (IE-MS) have been used for localized delivery of anticancer drugs and chemosensitizers. Because of their hydrophilic nature, the IE-MS release their payload quickly. The purpose of this work was to develop an IE-MS system that could provide a broad range of release rates for in vitro and in vivo applications. Sulfopropylated dextran microspheres (SP C25 MS) were surface-modified by acylation. These hydrophobically modified sulfopropylated dextran microspheres (HM-MS) were further coated with the cationic acrylic polymer Eudragit RL100 (EU-MS). The changes in chemical composition after the surface modification and coating were characterized by X ray photoelectron spectroscopy. The effects of the modification and coating on the surface hydrophobicity, equilibrium swelling, surface morphology, and drug loading capacity were investigated. The HM-MS showed little change in swelling and functionality, despite significantly increased affinity to oil and carbon content on the surface. The coated microspheres (EU-MS) exhibited a profoundly decreased swelling ratio, an even higher affinity to oil, a higher loading capacity, and a lower drug release rate. Through further coating of the EU-MS with different amounts of corn oil, the rate of drug release could be tailored to cover a relatively wide range. These results suggest that a versatile delivery system with various release profiles can be prepared by a combination of hydrophobic modification, polymer coating, and oil coating. PMID- 10824141 TI - The effects of a histidine residue on the C-terminal side of an asparaginyl residue on the rate of deamidation using model pentapeptides. AB - The effects of a histidine (His) residue located on the C-terminal side of an asparaginyl (Asn) residue on the rate of deamidation were studied using Gly-Gln Asn-X-His pentapeptides. The rates of deamidation of the pentapeptides were determined at 37 degrees C (I = 0.5) as function of pH, buffer species, and buffer concentration. A capillary electrophoresis stability-indicating assay was developed to monitor simultaneously the disappearance of the starting peptides and the appearance of the degradation products. The rates of degradation of the peptides were pH dependent, increasing with pH, and followed apparent first-order kinetics. At pH values <6.5, Gly-Gln-Asn-His-His degraded faster than Gly-Gln-Asn Gly-His, suggesting that the N+1 His residue is catalyzing the deamidation of the Asn residue. The His side chain at these pH values could function as a general acid catalyst, stabilizing the oxyanionic transition state of the cyclic imide intermediate formation. In contrast, at pH values >6.5, Gly-Gln-Asn-Gly-His deamidates more rapidly than Gly-Gln-Asn-His-His. The bulk of the side chain of the N+1 His residue versus the N+1 Gly residue apparently inhibits the flexibility of the peptide around the reaction site and, consequently, reduces the rate of the reaction. The significance of this steric hindrance effect of the N+1 His residue on the rate of deamidation was examined further. It was observed that at pH >6.0, Gly-Gln-Asn-His-His undergoes deamidation faster than Gly-Gln Asn-Val-His. This observation indicated that, at the higher pH values, the N+1 His residue is also acting as a catalyst. Thus, at basic pH, the N+1 His residue influences the rate of deamidation via two opposing effects; that is, general base catalysis and steric interference. The pentapeptide Gly-Gln-Asn-His-His, in addition to undergoing the deamidation reaction, also undergoes bond cleavage at the Asn-His peptide bond. The enhanced rate of Asn-His peptide bond cleavage can be attributed to the general base behavior of the His residue, leading to increased nucleophilicity of the Asn side-chain amide group. Finally, we have shown that the His residue that is two amino acids removed from the Asn, the N+2 position, has little or no effect on the rate of deamidation. PMID- 10824144 TI - Catalytic diastereoselective sulfimidation of diaryl sulfides and application of chiral sulfimides to asymmetric allylic alkylation. AB - The copper-catalyzed diastereoselective imidation of diaryl sulfides bearing a chiral oxazolinyl moiety at the ortho-position with [N-(p-toluenesulfonyl) imino]phenyliodinane (TsN=IPh) or Chloramine-T trihydrate [TsN(Cl)Na.3H2O] was successfully carried out to give the corresponding optically active N tosylsulfimides in good yields. For example, the imidation of diphenyl sulfide bearing a methoxymethyl moiety at the 4-position of the oxazoline ring with TsN(Cl)Na.3H2O in acetonitrile in the presence of 10 mol% Cu(OTf)2 at 25 degrees C for 24 h affords the corresponding optically active N-tosylsulfimide in 52% isolated yield with a high diastereoselectivity of 99%. Hydrolysis of the optically active N-p-tosylsulfimides converts them into the corresponding optically active sulfimides in high yields without loss of diastereoselectivity. These novel optically active sulfimides and N-tosylsulfimides work as efficient chiral ligands for palladium(II)-catalyzed allylic alkylation of 1, 3-diphenyl-3 acetoxy-1-propene with dimethyl malonate to give the corresponding alkylation product quantitatively and with a high stereoselectivity (up to 90% ee). PMID- 10824142 TI - Transport of cosalane-a highly lipophilic novel anti-HIV agent-across caco-2 cell monolayers. AB - Cosalane is a potent inhibitor of HIV replication with activity against a broad range of viral targets. However, the oral bioavailability of this highly lipophilic compound is extremely poor (<1%). Also, cosalane accumulates in high concentration in the liver after intravenous administration, with clear resistance to hepatic metabolism. In the present study, the transcellular permeability of cosalane was examined using Transwell(R) filter as well as plastic-grown confluent Caco-2 cell monolayers. A cell-culture-based biophysical model was adopted to understand the interactions of protein binding, membrane partitioning, and aqueous solubility of cosalane in limiting transcellular flux of cosalane across Caco-2 cell monolayers. The transcellular permeability (P(app)) of cosalane was extremely low (4.494 x 10(-8) cm/s) and the effect of p glycoprotein on the efflux of cosalane was negligible. A characteristic disparity exists between the kinetics of cosalane uptake from apical (AP) donor solution and efflux into basolateral (BL) receiver side. The AP uptake of cosalane was rapid, exhibiting exponential kinetics, and reached equilibrium within 60 min, whereas the concomitant appearance of the compound into the BL receiver side was slow but linear over time. Furthermore, the uptake of cosalane was significantly reduced in the presence of bovine serum albumin (BSA). In unidirectional efflux studies, AP efflux of cosalane was limited in the absence of BSA. Also, no detectable metabolites were found in Caco-2 cell incubations. In conclusion, the present study demonstrates that diffusion of cosalane across Caco-2 cell monolayers is extremely limited and kinetically regulated essentially by the equilibrium between protein-bound and free drug partitioning into cell membrane. PMID- 10824145 TI - PaaSiCats: novel polyamino acid catalysts. AB - The abilities of five polyamino acids (Paa's) to catalyse the asymmetric epoxidation of enones 1-7 under three sets of reaction conditions were compared: polyneo-pentylglycine and polyleucine showed distinct advantages in most circumstances. All five polymers were adsorbed onto silica and from this further study, immobilised polyneo-pentylglycine (PLNSi) and polyleucine (PLLSi) were shown to be the catalysts of choice for the asymmetric epoxidation of less reactive alpha,beta-unsaturated ketones. PMID- 10824146 TI - First synthesis and resolution of a planar-chiral tetrahydroindolyl complex of iron: electronic tuning of reactivity and enantioselective nucleophilic catalysis. AB - The first examples of an (eta5-indolyl)iron complex and of an (eta5 tetrahydroindolyl)iron complex are described. Reactivity studies establish that the (eta5-tetrahydroindolyl)iron complexes are the most active azaferrocene derived nucleophilic catalysts reported to date and that the reactivity of these complexes can be electronically tuned. Use of planar-chiral, enantiopure (eta5-3 (dimethylamino)tetrahydroindolyl)FeCp* in asymmetric catalysis leads to stereoselectivities comparable to those furnished by a previously described azaferrocene complex. PMID- 10824147 TI - Diastereoselectivity an chiral ruthenium complexes of chiraphos monoxide ligands: epimerization, quasiracemates, and Lewis acid catalysis. AB - Heterobidentate chelating ligands comprising P,O-donor sets produce chiral metal centers when bound to arene-ruthenium complexes. The cymene complex [CyRu(eta2 chelate-P,O)Cl]+ cations are precursors to 16-electron dicationic strong Lewis acids which have potential use in asymmetric catalysis. Sixteen-electron complexes of this type, however, also provide a pathway with a low energy barrier to epimerization of the metal center in intermediates. With the chiral ligand (R,R)-chiraphos monoxide, the metal center in [CyRu(eta2)-(R,R)-Ph2PCHMeCHMe Ph2PO-P,O)Cl]+ epimerizes to minimize steric interactions in the two diastereomers yielding a 8. 2:1 dr in solution. Nevertheless, upon crystallization a new equilibrium is obtained in the solid and the crystals obtained have a 1:1 dr. These crystals are quasiracemates, which contain equal amounts of the (RRu) and (SRu)-CyRu-P-O-Cl stereocenters within the same crystal. The doubly charged Lewis acid, [CyRu(eta2-(R, R)-Ph2PCHMeCHMe Ph2PO P,O)(solvate)]2+ derived from the chloro complex by chloride abstraction with AgSbF6 gave modest ee's (30%) in the Diels-Alder reaction of methacrolein with cyclopentadiene. PMID- 10824148 TI - Integrated route to the L-aldohexoses using a common man-made chiral building block. AB - Starting from a common man-made chiral sugar building block, five out of eight L aldohexoses have been synthesized in a diastereocontrolled manner to complete an integrated synthesis of the eight possible aldohexoses; the remaining three L hexoses were synthesized from the same building block. Thus, L-altrose, L mannose, L-allose, L-talose, and L-glucose were obtained in the present investigation on the basis of the inherent convex-face selectivity exerted by the chiral building block. PMID- 10824149 TI - Asymmetric synthesis of functionalized carbocycles and heterocycles. AB - Anions of 1-halo-4-hexenyl phosphonamides derived from chiral, enantiopure C2 symmetrical 1,2-diamino cyclohexane react at the gamma-position in conjugate addition reactions with alpha, beta-unsaturated carbonyl compounds such as cyclopentenone, 4-(H)-furanone, pyrroline-2-one, and cinnamates to give functionalized adducts. Addition to imines is also possible. The adducts can be transformed into enantiopure or enriched carbocyclic and heterocyclic compounds bearing useable functionality. PMID- 10824150 TI - Synthesis of a new enantiomerically pure P-chiral phosphine and its use in probing the mechanism of the Mitsunobu reaction. AB - A new enantiomerically pure P-chiral phosphine, (S)-cyclohexylmethyl- (1 naphthyl)phosphine (1) was prepared by phosphine-borane methodology and used in a mechanistic study of the Mitsunobu reaction. Enantiomerically enriched (S)-cyclo- hexylmethyl(1-naphtyl)phosphine oxide (8) is obtained if the reaction proceeds through the phosphonium salt 4, whereas the intermediate dialkoxyphosphorane 5 leads to racemic phosphine oxide 8. The results of the experiments including the variation of the reaction conditions and the natures of alcohol and carboxylic acid used in the Mitsunobu reaction prove the competition of two alternative mechanisms (reaction via 4 or 5) on the second stage of the Mitsunobu reaction. PMID- 10824151 TI - Synthesis of axially chiral benzamides utilizing tricarbonyl(arene)chromium complexes. AB - Axially chiral N,N-diethyl 2,6-disubstituted benzamides were stereo-selectively prepared utilizing planar chiral (arene)chromium complexes as an enantiomerically active form by following two methods. Ortho-lithiation of the enantiomerically pure planar chiral tricarbonyl(N,N-diethyl 2-methylbenzamide)chromium complex followed by electrophilic quenching gave axially chiral 2-methyl-6-substituted N,N-diethyl benzamide chromium complexes. Photo-oxidative demetalation produced the chromium-free axially chiral benzamides as optically active compounds. An alternative method for the preparation of axial chiral benzamides is an enantioselective lithiation at the benzylic methyl of meso tricarbonyl(N,N diethyl 2,6-dimethylbenzamide)chromium with appropriate chiral lithium amide base followed by quenching with alkyl halides. PMID- 10824152 TI - Radical cyclization of beta-alkoxyacrylates: synthesis of a C2-symmetric, L shaped molecule with four fused tetrahydropyran rings. AB - A C2-symmetric, L-shaped tetracycle with four fused tetrahydropyran rings was synthesized from (3R, 4R)-1, 6-dibenzoyloxy-hexane-3, 4-diol employing radical cyclization reactions of beta-alkoxyacrylates. PMID- 10824153 TI - Enantioresolution by the chiral phthalic acid method: absolute configurations of substituted benzylic alcohols. AB - Substituted benzylic alcohols were enantioresolved by the chiral phthalic acid method as follows; 1) esterification of racemic alcohols with chiral phthalic acid, 2) separation of a diastereomeric mixture of the esters formed by HPLC on silica gel, and 3) recovery of enantiopure alcohols from the separated esters. The absolute configurations of chiral phthalic acid esters of benzylic alcohols were unambiguously determined by the X-ray crystallography using the campharsultam moiety as the internal standard of absolute configuration. PMID- 10824154 TI - Stereoconvergent approach for synthesizing enantiopure 5, 6-dialkylpipecolic acids. AB - Investigating a general route for synthesizing pipecolic acid ) piperidine-2 carboxylic acid ( derivatives with substituents at the 3-, 4-, 5- and 6-position, we discovered a stereoconvergent process that provides an effective means for making 5, 6-dialkyl-epsilonpipecolate (Scheme 1, PhF = 9-phenylfluoren-9-yl). Hydrogenation of diastereomeric mixtures of gamma-oxo gamma-hydroxy and gamma acetoxy alpha-N-(PhF)amino tert-butyl esters causes the eventual loss of the gamma-substituent to furnish an azadiene intermediate that can reduce diastereoselectively to 5, 6-dialkylpipecolate having the all cis relative stereoconfiguration. Five enantiopure (>94% ee) 5,6-dialkylpipecolic acids were synthesized, employing aspartic acid as an inexpensive chiral educt in this process. PMID- 10824155 TI - Efficient asymmetric synthesis of alpha-alkylated 1, 4-cyclohexanedione derivatives, important chiral building blocks in the synthesis of natural products. AB - Alkylation of 1,4-cyclohexanedione monoethylene acetal SAMP-hydrazone with various electrophiles (10 examples given) and subsequent cleavage of the hydrazones with saturated oxalic acid furnished highly enantiomerically enriched alpha-alkylated mono-protected 1,4-cyclohexanedione derivatives in high yields and enantiomeric excesses of ee = 28, 80->/=99%. Reduction of the ketones gave the corresponding alcohols in good yields with high enantiomeric excesses (ee = 80->/=98%) and cis/trans-ratios of usually 85:15. PMID- 10824156 TI - Asymmetric modular synthesis of cylindrically chiral FerroPHOS ligands for the Rh catalyzed asymmetric hydroboration. AB - Asymmetric modular synthesis of air-stable ferrocenyl bisphosphine ligands with cylindrical chirality was achieved employing asymmetric catalytic methods and these chiral ligands were screened in asymmetric hydroboration reaction. PMID- 10824158 TI - Preparation of new chiral borane-protected P,N-ferrocenyl ligands via a methoxy directed ortho-lithiation. AB - We have shown that the readily prepared (ferrocenyl)benzylic methyl ethers of type 1 can be ortho-metalated with tert-BuLi with high diastereoselectivity. This reaction has been used to prepare new borane-protected P,N-substituted ferrocenes of type 2 in high diastereomeric and enantiomeric purity. The chiral heterocyclic aminophosphines 2 may be of interest as chelating ligands for asymmetric metal catalysis. PMID- 10824157 TI - New chiral catalysts for reduction of ketones. AB - The condensation of o-(diphenylphosphino)benzaldehyde and various chiral diamine gives a series of diimino-diphosphine tetradentate ligands, which are reduced with excess NaBH4 in refluxing ethanol to afford the corresponding diaminodiphosphine ligands in good yield. The reactivity of these ligands toward trans-RuCl2(DMSO)4 and [Rh(COD)Cl]2 had been investigated and a number of chiral Ru(II) and Rh(I) complexes with the PNNP-type ligands were synthesized and characterized by microanalysis and IR, NMR spectroscopic methods. The chiral Ru(II) and Rh(I) complexes have proved to be excellent catalyst precursors for the asymmetric transfer hydrogenation of aromatic ketones, leading to optically active alcohols in up to 97% ee. PMID- 10824159 TI - Unusual solvent effects on chiroptical properties of an optically active regioregular polythiophene in solution. AB - The chiroptical properties of a novel, optically active regioregular poly[3-(4 ((R)-4-ethyl-2-oxazolin-2-yl)phenyl)thiophene] (poly-1) were investigated in mixtures of a good solvent, chloroform, and a variety of poor solvents using CD spectroscopy. Most poor solvents induced CD on poly-1 with similar Cotton effects, while acetonitrile and nitromethane caused dramatic changes in the Cotton effects of poly-1. PMID- 10824160 TI - Catalytic asymmetric synthesis of 19,20-dihydroakuammicine. AB - An enantiocontrolled total synthesis of 19,20-dihydroakuammicine using a catalytic asymmetric Michael addition of dimethyl malonate to cyclohexenone as the key step is described. The above catalytic asymmetric Michael addition proceeds quite efficiently in the presence of a heterobimetalic asymmetric catalyst (ALB-KO-t-Bu-MS 4A, 0.3 mol%), giving the corresponding Michael adduct in 94% yield and 99% ee. PMID- 10824161 TI - Asymmetric synthesis of 1,2-amino alcohols using (S)-indoline chiral auxiliary. AB - Chiral hydrazones 1 reacted with aryl- or alkyl-lithiums at -78 degrees C in a short reaction time, within 10 min, to afford arylated or alkylated chiral hydrazines 3 with extremely high diastereoselectivity (up to >99% de) and high chemical yields. The hydrazines are readily converted to chiral amino alcohols 4. PMID- 10824162 TI - Asymmetric synthesis of hydroxylated pyrrolizidine, indolizidine, and (+)-alpha conhydrine via ruthenium-catalyzed hydrogenation. AB - The enantioselective ruthenium promoted hydrogenation of beta-keto ester, derived from (S)- or (R)-proline and (S)-pipecolic acid, provided a new efficient route to hydroxylated pyrrolizidine or indolizidine ring systems in diastereomeric excesses up to 99%. A practical synthesis of (+)-alpha-conhydrine is also reported. PMID- 10824163 TI - Immobilization of rhodium complexes in chiral organic-inorganic hybrid materials. AB - Two new alkoxysilylated derivatives of (-)-(1R,2R)-1, 2-diaminocyclohexane: M = N [(triethoxysilyl)propyl]-(-)-(1R,2R)-1, 2-diaminocyclohexane and B = N, N' bis[(triethoxysilyl)propyl]-(-)-(1R,2R)-1,2-diaminocyclohexane have been synthesized. Their complexation with [Rh(cod)Cl]2 in the presence of TEOS = Si(OEt)4, followed by sol-gel hydrolysis-condensation, afforded new catalytic chiral hybrid materials. Evidence for the presence of the organic moieties complexed by rhodium in these solids was obtained by UV-visible spectroscopy, FT IR studies, solid state 13C and 29Si CP-MAS NMR analysis, energy-dispersive X-ray (EDX) techniques, and elemental analysis. The nitrogen sorption studies and BET analyses ranged these solid gels from nonporous to highly porous materials. The catalytic activities and selectivities of the solid materials have been studied in the asymmetric hydrogen-transfer reduction of prochiral ketones and compared to that of the homogeneous rhodium complexes of the ligands M and B. The hybrid materials appeared interesting supports for enantioselective heterogeneous catalysis leading to chiral alcohols with ee up to 58% in the reduction of acetophenone and up to 98% in the case of the more hindered related ketones. The catalytic properties as a function of the nature of chiral hybrid solid are discussed. PMID- 10824164 TI - Asymmetric aldol reaction of enol trichloroacetate catalyzed by (S, S) (EBTHI)TiCl(OMe). AB - (S,S)-Ethylenebis(tetrahydroindenyl)titanium chloride methoxide, (S, S) (EBTHI)TiCl(OMe) (3) was synthesized from the corresponding titanium dichloride. The asymmetric aldol reaction of enol trichloroacetate of cyclohexanone 1 with aromatic aldehydes was studied in the presence of a catalytic amount of the chiral titanium complex 3, with the result that the optically active syn aldol adduct 2 was preferentially obtained with up to 91% ee. PMID- 10824165 TI - Practical stereoselective synthesis of (2,3,4, 5-tetrahydro-1H-benzo[b]azepin-5 yl)acetic acid. AB - Highly enantioselective asymmetric hydrogenation of readily accessible olefins, (E)- and (Z)-[1-(toluene-4-sulfonyl)-1,2,3, 4-tetrahydro-1H-benzo[b]azepin-5 ylidene]acetic acid (4a and 4b, respectively) and [1-(toluene-4-sulfonyl)-2, 3 dihydro-1H-benzo[b]azepin-5-yl]acetic acid (4c), is presented as an efficient and straightforward route to (R)-[1-(toluene-4-sulfonyl)-2,3,4, 5-tetrahydro-1H benzo[b]azepin-5-yl]acetic acid [(R)-1] which is a key intermediate for the synthesis of non-peptide AVP V2-agonist. Hydrogenation of carboxylic acid 4c gave (R)-1 in quantitative yield and 85% ee using Ru(OAc)2[(S)-H8-BINAP], a Ru(II) complex of a partially hydrogenated BINAP (H8-BINAP), as a catalyst. When (R)-1 of 76% ee was transformed into the corresponding isopropylamide 6, pure enantiomer (R)-6 was obtained in 75% yield by recrystallization from MeOH. PMID- 10824166 TI - Synthesis of highly potent second-generation taxoids through effective kinetic resolution coupling of racemic beta-lactams with baccatins. AB - A series of highly potent second-generation taxoids bearing a 2-methylprop-1-enyl or a 2-methylpropyl group at C-3' with modifications at the C-2, C-10, and C-14 positions was synthesized through the coupling of racemic cis-beta-lactams with properly protected/modified baccatin and 14-OH-baccatin. A high level of kinetic resolution was observed for all cases examined. The observed highly efficient enantiomer differentiation is ascribed to the markedly different chiral environment between the (+)- and (-)-beta-lactams in their approach to the chiral framework of the enantiopure lithium alkoxide of a baccatin in the ring-opening coupling process. It was also observed that substantially higher selectivity was achieved when 14-OH-baccatin-1,14-carbonate was used. Analysis of the transition state models revealed that the repulsive interactions between the 3-TIPS group of a (-)-beta-lactam with 1, 14-carbonate group of the baccatin substantially increases the asymmetric bias in the kinetic resolution process, favoring the reaction of a (+)-beta-lactam, which leads to the observed excellent selectivity. PMID- 10824167 TI - Enantioselective hydrogenation of olefins with phosphinooxazoline-iridium catalysts. AB - Cationic iridium complexes with chiral phosphinooxazoline ligands are efficient catalysts for the enantioselective hydrogenation of olefins. The complexes are readily prepared, air-stable, and easy to handle. In contrast to chiral rhodium- and ruthenium-phosphine catalysts, they do not require the presence of a polar coordinating group near the C=C bond. In the hydrogenation of unfunctionalized trisubstituted 1,2-diaryl-olefins, high enantioselectivities of >95% ee with full conversion and turnover frequencies of >7,000 h-1 can be achieved, using 0.1 mol% of catalyst with tetrakis[3, 5-bis(trifluoromethyl)phenyl]borate (TFPB or BARF) as the counterion. The corresponding hexafluorophosphate or tetrafluoroborate salts give low conversion due to deactivation of the catalyst during the reaction. Substrates with polar substituents such as allylic alcohols, on the other hand, afford better results with the hexafluorophosphate salts. PMID- 10824168 TI - Mechanistic aspects of dihydrogen activation and transfer during asymmetric hydrogenation in supercritical carbon dioxide. AB - A new "CO2-philic" chiral rhodium diphosphinite complex was synthesized and applied as catalyst precursor in the asymmetric hydrogenation of dimethyl itaconate in scCO2, scC2H6 and various liquid organic solvents. Deuterium labeling studies and parahydrogen-induced polarization (PHIP) NMR experiments were used to provide the first detailed mechanistic insight into the activation and transfer of the dihydrogen molecule during hydrogenation in scCO2. Chemical interactions between CO2 and reactive intermediates of the catalytic pathway could be excluded as possible explanations for the experimentally verified difference in the catalytic behavior in scCO2 and hexane. PMID- 10824169 TI - New efficient chiral derivatizing agent, alpha-cyano-alpha-fluoro(2 naphthyl)acetic acid (2-CFNA). application to the EE determination of (-)-3 acetoxy-2-fluoro-2-(hexadecyloxymethyl)propan-1-ol. AB - The new chiral derivatizing agent (CDA), alpha-cyano-alpha-fluoro(2-naphthyl) acetic acid (2-CFNA) 1 was prepared in optically pure form by chiral HPLC separation of racemic 2-CFNA methyl ester (2-CFNA Me ester) (+/-)-2. The ester was obtained by fluorination of methyl alpha-cyano(2-naphthyl)acetate with FClO3. 2-CFNA 1 has proven to be a significantly superior CDA for determination of enantiomeric excess (ee) of a primary alcohol when compared to alpha-methoxy alpha-trifluoromethylphenylacetic acid (MTPA, Mosher's agent) and alpha-cyano alpha-fluoro(p-tolyl)acetic acid (CFTA). The ee of (-)-3-acetoxy-2-fluoro-2 (hexadecyloxymethyl)propan-1-ol (-)-9, a fluorinated analog of anticancer active ether lipids, was determined using (+)-2-CFNA (+)-1. PMID- 10824170 TI - Short step enantioselective synthesis of (-)-swainsonine. AB - Recent discovery of asymmetric desymmetrization of meso-heterocycles using (R,R) (salen)manganese complex 2 as a catalyst allowed easy access to optically active 2,3,4-trisubstituted pyrrolidines. Taking advantage of this new reaction, we could achieve the short-step synthesis of (-)-swainsonine 1. PMID- 10824171 TI - Asymmetric 1,4-addition of phenylboronic acid to 2-cyclohexenone catalyzed by Rh(I)/binap complexes. AB - Reaction of 2-cyclohexenone with phenylboronic acid in the presence of 3 mol% of a rhodium(I)/(S)-binap catalyst in dioxane/H2O (10/1) at 100 degrees C proceeded with high enantioselectivity to give a high yield of (S)-3-phenylcyclohexanone of up to 99% ee. The high enantioselectivity was achieved by use of a catalyst generated in situ from Rh(acac)(C2H4)2 and (S)-binap or Rh(acac)((S)-binap) as an isolated rhodium-phosphine complex. PMID- 10824172 TI - Optically active titanium complexes containing a tridentate linked amido cyclopentadienyl ligand. AB - Optically active titanium complexes Tieta5:eta1-C5R4SiMe2NC6H10 (OCH2Ph)-2Cl2 (R = H, Me), containing a cyclopentadienyl ligand linked to the chiral trans-2 benzyloxycyclohexylamido group, were synthesized and characterized in both enantiomerically pure forms. A single crystal X-ray structure analysis of (-)-(R, R)-Tieta5:eta1-C5H4SiMe2NC6H10(OCH2Ph)-2Cl2 shows a structure in which the benzyloxy group in the amido sidechain is not interacting with the titanium center. Upon activation with n-butyllithium, these complexes hydrogenate acetophenone N-benzylimine with low enantioselectivity. PMID- 10824173 TI - Kinetic resolution and isomerization of 2,5-disubstituted pyrrolines. AB - The efficient kinetic resolution of 2,5-disubstituted pyrrolines was accomplished by employing hydrosilylation with a chiral catalyst, (EBTHI)TiF2 (EBTHI = ethylenebis(tetrahydroindenyl)). An interesting isomerization reaction of the cyclic imines catalyzed by the same chiral catalyst was also discovered. PMID- 10824174 TI - Circular dichroism as a detection method in the screening of enantioselective catalysts. AB - The combination of liquid chromatography (HPLC), UV/Vis-spectroscopy and circular dichroism (CD) can be used to construct a high-throughput screening system to determine the enantioselectivity of enzyme- or metal-catalyzed reduction of acetophenone with formation of (S)- and (R)-1-phenylethanol. Prerequisite for the viability of this system is the experimental finding that the anisotropy factor g is linearly related to the enantiomeric excess (ee) and that it is independent of concentration, thereby excluding possible aggregation effects. PMID- 10824175 TI - Kinetic resolution of alpha-acetoxy carboxylic acids with homochiral SuperQuats. AB - Preliminary investigations focused on the kinetic resolution of (+/-)-alpha acetoxy mandelic carboxylic acid chloride with isopropyl SuperQuat and isopropyl Evans' homochiral auxiliaries. The addition of 2 equiv. of the acid chloride to the lithium salts of the auxiliaries (SuperQuat and Evans') at -100 degrees C in the THF afforded the corresponding N-acyl auxiliaries in high chemical yields (95% and 89%) and d.e.'s of 66% and 40%, respectively. Using the same mandelic acid chloride but incorporating SuperQuat auxiliaries derived from (S) phenylglycine and (S)-phenylalanine yielded the corresponding N-acyl SuperQuats in 86% and 90% and d.e. 's of 66% and 30%, respectively. Substituting the phenyl group within the alpha-acetoxy mandelic acid chloride for a n-butyl, tert-butyl and cyclohexyl group changed the d.e. significantly when kinetically resolved with isopropyl SuperQuat. The yields were all similar, i.e., approximately 90% but the d.e.'s varied considerably, with values of 20%, 50%, and 82%, respectively. Attempted kinetic resolution of alpha-methoxyphenylacetyl chloride using the lithium salt of isopropyl SuperQuat revealed a diasteroselectivity of 45%, significantly lower than that obtained with the corresponding alpha-acetoxy carboxylic acid chlorides. PMID- 10824176 TI - IMDA/aldol strategy for transforming carbohydrates into functionalized trans decalins. AB - L-Rhamnal is readily converted into an allyl 2, 3-unsaturated-C-glycopyranoside. The (S) configuration of the alphaL-anomer defines the stereochemical outcome of the future IMDA reaction, leading to the absolute stereochemistry for the trans decalin moiety in naturally occurring terpenoids. Selective cleavage of the terminal double bond of the allyl group provides an aldehydo function which serves for an aldol/Claisen addition with ethyl sorbate. Of the four possible diastereomers, one is obtained in pure form and processed to give the IMDA precursor. Cyclocondensation is achieved by heating in xylene to give a tricyclic trans-decalin whose structure is established by NMR and X-ray analysis. PMID- 10824177 TI - Diastereoselectivity in scalemic tartrate/titanium epoxidations. AB - Nonlinearity in the diastereoselectivity of epoxidation of allylic alcohols with mixtures of titanium isopropoxide, tertbutyl hydroperoxide, and diethyl tartrate was observed. Racemic and enantiomerically pure alcohols E-2-methyl-4-hexen-3-ol and E-1-methoxy-5-(O-tertbutyldimethylsilyloxy)-2-penten-4-ol were prepared. Epoxidation reactions were carried out with Ti(OPri)4 and ButOOH accompanied by diethyl tartrate of varying enantiomeric purity. The simplest explanation of these results is that a dimeric epoxidation reagent is involved, with significantly different reactivity for the homochiral and racemic forms. PMID- 10824178 TI - External chiral ligand-induced enantioselective versions of the [2,3]-Wittig sigmatropic rearrangement. AB - The external chiral ligand-induced enantioselective [2,3]-Wittig rearrangements of crotyl benzyl ethers and crotyl propargylic ethers are described. The most notable is that treatment of (E)-crotyl propargylic ethers with a t butyllithium/(S;S)-bis(oxazoline) complex provides a relatively high enantioselectivity (up to 89% ee), together with a high threo diastereoselectivity. Furthermore, examples of the "asymmetric catalytic version" of the rearrangement of crotyl benzyl ethers are presented. PMID- 10824179 TI - Synthesis of a new chiral ligand, 6, 6'-dihydroxy-5, 5'-biquinoline (BIQOL) and its applications in the asymmetric addition of diethylzinc to aldehydes. AB - A new chiral ligand 6, 6'-dihydroxy-5, 5'-biquinoline (BIQOL, 2) was prepared via Cu2+ mediated coupling. The resolution was carried out by separating the corresponding ditrifluomethanesulfonate on chiral column. When applied to the enantioselective addition of diethylzinc to aromatic aldehydes, this ligand induced the reaction with enantioselectivity equivalent to that induced by BINOL. The effects of solvent and reaction temperature on enantioselectivity were also studied. PMID- 10824180 TI - Versatile precursor to ruthenium-bis(phosphine) hydrogenation catalysts. AB - The known complex trans-RuCl2(NBD)Py2 (1, NBD is norbornadiene, Py is pyridine) reacts with either (R)-BINAP ((R)-2, 2'-bis(diphenylphosphino)-1,1'-binaphthyl), (S;S)-Chiraphos ((2S;3S-(-)-2,3-bis(diphenylphosphino)butane), (S;S)-Skewphos ((2S;4S)-(-)-2,4-bis(diphenylphosphino)pentane), (R)-(S)-Josiphos ((R)-(-)-1-[(S) 2-(diphenylphosphino)ferrocenyl]ethyl-dicyclohexylpho sphine), (R;R)-Norphos ((2R;3R)-(-)-2, 3-bis(diphenylphosphino)bicyclo[2.2.1]hept-5-ene), or (R;R)-Me DUPHOS ((-)-1,2-bis((2R;5R)-2, 5-dimethylphospholano)benzene) to generate in high yields the crystalline complexes trans-RuCl2(P-P*)Py2 (P-P* is the corresponding chiral bis(phosphine)). The complexes trans-RuCl2(P-P*)Py2 are active enantioselective hydrogenation catalysts for ketoesters and noncarboxylic olefins in the presence of small amounts of HBF4 (aq.). They are active for hydrogenation of carboxylic substrates in the presence of Et3N. Reaction of trans-RuCl2(P P*)Py2 with (rac)-1,2-diphenylethylene-diamine (N-N*, either enantiomer) forms in good yields the corresponding compounds trans-RuCl2(P-P*)(N-N*). Representative hydrogenations with these catalysts are presented. PMID- 10824181 TI - Zirconium-mediated asymmetric baeyer-villiger oxidation. AB - Combinations of axially chiral C2-symmetric diols were used as ligands in zirconium-mediated Baeyer-Villiger reactions. The in situ preformed Zr-diol species proved effective in the asymmetric oxidation of bicyclic and monosubstituted cyclobutanones when a hydroperoxide was employed as oxidant. Asymmetric induction could be preserved upon replacement of one out of two enantiopure BINOL ligands by conformationally flexible 2,2'-biphenol. PMID- 10824182 TI - Transition metal complexes in organic synthesis, part 59.(1) First enantioselective total synthesis of lavanduquinocin, a potent neuronal cell protecting substance from streptomyces viridochromogenes. AB - Using (R)-propene oxide as a chiral building block a convergent enantioselective synthesis of the potent neuronal cell protecting alkaloid lavanduquinocin has been accomplished by the iron-mediated one-pot construction of the carbazole framework. PMID- 10824183 TI - alpha-t-butyl- and alpha-i-propyl-ortho-hydroxybenzylamines: racemic synthesis/resolution and asymmetric synthesis. AB - Efficient routes to alpha-tert-butyl- and alpha-iso-propyl-ortho hydroxybenzylamines 1a and 1b are described. Highly enantioenriched 1a and 1b were obtained by resolution of the methoxy derivatives 2 by recrystallization of the salts formed with mandelic acid followed by Lewis acid mediated demethylation. The chiral 1,3-amino alcohol 1a has also been obtained in an asymmetric synthesis with the key step a diastereoselective alkylation of the imine obtained by condensation of o-anisaldehyde with phenyl glycinol. The absolute stereochemistry of these 1,3-aminophenols was determined by CD spectroscopy of the salicylideneamines 12 and by an X-ray structure analysis of the salt formed between (R)-mandelic acid and (S)-alpha-tert-butyl-ortho methoxybenzylamine ((S)-2a). PMID- 10824184 TI - Novel binuclear chiral zirconium catalysts used in enantioselective strecker reactions. AB - A novel binuclear chiral zirconium catalyst was successfully used in enantioselective Strecker reactions. The catalyst was readily prepared from zirconium t-butoxide (Zr(OtBu)4), (R)-6,6'-dibromo-1, 1'-bi-2-naphthol ((R)-6-Br BINOL), and (R)-3,3'-dibromo-1, 1'-bi-2-naphthol ((R)-3-Br-BINOL) to form unique binuclear structure. It was revealed that a combination of (R)-6-Br-BINOL and (R) 3-Br-BINOL was essential in these asymmetric reactions and that much lower selectivities were obtained by using other combinations. Two-component (an imine and hydrogen cyanide (HCN)) and three-component (an aldehyde, an amine, and HCN) Strecker reactions proceeded smoothly in the presence of a catalytic amount of the chiral zirconium catalyst to afford the corresponding alpha-amino nitrile derivatives in high yields with high enantioselectivities. PMID- 10824185 TI - Optical rotation per refractive index unit, or enantiomeric (e) factor, for screening enantioselective catalysts through asymmetric activation or carbohydrates. AB - A super high-throughput screening (SHTS) system can be constructed by combining high-performance liquid chromatography (HPLC), optical rotation (OR), and refractive index unit (RIU) to determine not only the enantioselectivity of the addition of diethylzinc to an aliphatic aldehyde catalyzed by a binaphthol-zinc complex through asymmetric activation with chiral Schiff bases, but also the enantiopurity of a carbohydrate. The enantiomeric (e) factor, which we define here as optical rotation per refractive index unit, is linearly related to the percent enantiomeric excess (%ee) and is independent of concentration. PMID- 10824186 TI - Thermodynamic and kinetic properties of trans-3,4,5,6-tetramethyl-9, 10-dihydroxy 9,10-dihydrophenanthrene: pseudo-rotational barriers around biphenyl axis. AB - The thermodynamic and kinetic properties of trans-3,4,5, 6-tetramethyl-9,10 dihydro-9,10-dihydroxyphenanthrene (4) and its derivatives were studied. The equilibrium ratios of two the possible conformers (4eq, 4ax) were measured in various solvents, where consistently the equatorial conformers proved to be more stable than those of the corresponding axial counterparts. The activation barriers (DeltaG) for the conformational change, 4ax-->4eq, were roughly estimated to be ca. 26 kcal mol-1 at 298 K in various solvents. PMID- 10824187 TI - Compensasome in Drosophila. PMID- 10824188 TI - Size control in development: lessons from Drosophila. PMID- 10824189 TI - Should erythrocyte destruction in vivo be through phagocytosis alone? PMID- 10824190 TI - Transfer of learning across the somatosensory cortex. PMID- 10824191 TI - Molecular slavery. PMID- 10824192 TI - The smallest form of life yet? PMID- 10824193 TI - The sight of the peacock's tail makes me sick: the early arguments on sexual selection. AB - Why does a peacock have a beautiful train, while a peahen is sober without such flamboyance? Darwin proposed the theory of sexual selection to explain the differences between the sexes of the same species. Recently the study of sexual selection has been one of the most flourishing areas in evolutionary biology. However, the theory met with great resistance from biologists since the publication of the idea and the history of the theory included a lot of misunderstanding and confusion. There are several reasons for this. First, classical Darwinism failed to recognize social competition as an important selective force. Second, the good-for-the-species argument, which persisted in the days after Darwin, made the sexual selection argument more difficult to under stand. Compared to the discussions on animals, Darwin's argument on human sex differences is not satisfactory. The reason probably lies in the debate over human racial differences which prevailed in the 19th century. PMID- 10824194 TI - Analysis of BRCA1 involvement in breast cancer in Indian women. AB - The involvement of the familial breast-ovarian cancer gene (BRCA1) in the molecular pathogenesis of breast cancer among Indian women is unknown. We have used a set of microsatellite polymorphisms to examine the frequency of allele loss at the BRCA1 region on chromosome 17q21, in a panel of 80 human breast tumours. Tumour and blood leukocyte/normal tissue DNA from a series of 80 patients with primary breast cancer was screened by PCR-amplified microsatellite length polymorphisms to detect deletions at three polymorphic BRCA1 loci. PCR allelotype was valuable in examining allele losses from archival and small tumour samples. Loss of alleles at BRCA1 in the patient set, confirmed a noteworthy role of this gene in the molecular patho-genesis of breast cancer and was in accordance with its well-documented tumour suppressive function. PMID- 10824195 TI - Growth modulation of fibroblasts by chitosan-polyvinyl pyrrolidone hydrogel: implications for wound management? AB - Wounds in adults and fetuses differ in their healing ability with respect to scar formation. In adults, wounds lacking the epidermis exhibit excess collagen production and scar formation. Fibroblasts synthesize and deposit a collagen rich extracellular matrix. The early migration and proliferation of fibroblasts in the wound area is implicated in wound scarring. We have synthesized a hydrogel from chitosan-polyvinyl pyrrolidone (PVP) and examined its effect on fibroblast growth modulation in vitro. The hydrogel was found to be hydrophilic as seen from its octane contact angle (141.2+/-0.37 degrees). The hydrogel was non-toxic and biocompatible with fibroblasts and epithelial cells as confirmed by the 3(4,5 dimethylthiazolyl-2)-2, 5-diphenyl tetrazolium bromide (MTT) as-say. It showed dual properties by supporting growth of epithelial cells (SiHa) and selectively inhibiting fibro-blast (NIH3T3) growth. Growth inhibition of fibroblasts resulted from their inability to attach on to the hydrogel. These findings are supported by image analysis, which revealed a significant difference (P<0.05) between the number of fibroblasts attached to the hydrogel in tissue culture as compared to tissue culture treated polystyrene (TCPS) controls. However, no significant difference was observed (P>0.05) in the number of epithelial (SiHa) cells attached on to the hydrogel as compared to the TCPS control. Although in vivo experiments are awaited, these findings point to the possible use of chitosan-PVP hydrogels in wound-management. PMID- 10824196 TI - PTP-S2, a nuclear tyrosine phosphatase, is phosphorylated and excluded from condensed chromosomes during mitosis. AB - PTP-S2 is a tyrosine specific protein phosphatase that binds to DNA and is localized to the nucleus in association with chromatin. It plays a role in the regulation of cell proliferation. Here we show that the subcellular distribution of this protein changes during cell division. While PTP-S2 was localized exclusively to the nucleus in interphase cells, during metaphase and anaphase it was distributed throughout the cytoplasm and excluded from condensed chromosomes. At telophase PTP-S2 began to associate with chromosomes and at cytokinesis it was associated with chromatin in the newly formed nucleus. It was hyperphosphorylated and showed retarded mobility in cells arrested in metaphase. In vitro experiments showed that it was phosphorylated by CK2 resulting in mobility shift. Using a deletion mutant we found that CK2 phosphorylated PTP-S2 in the C-terminal non catalytic domain. A heparin sensitive kinase from mitotic cell extracts phosphorylated PTP-S2 resulting in mobility shift. These results are consistent with the suggestion that during metaphase PTP-S2 is phosphorylated (possibly by CK2 or a CK2-like enzyme), resulting in its dissociation from chromatin. PMID- 10824197 TI - Cloning of partial putative gonadotropin hormone receptor sequence from fish. AB - A search for the presence of mariner-like elements in the Labeo rohita genome by polymerase chain reaction led to the amplification of a partial DNA sequence coding for a putative transmembrane domain of gonadotropin hormone receptor. The amplified DNA sequence shows a high degree of homology to the available turkey and human luteinizing and follicle stimulating hormone receptor coding sequences. This is the first report on cloning such sequences of piscine origin. PMID- 10824198 TI - An enzyme immunoassay for detection of Japanese encephalitis virus-induced chemotactic cytokine. AB - Japanese encephalitis virus (JEV) induces human peripheral blood monocytes to secrete a chemotactic cytokine [human macrophage-derived factor (hMDF)] which causes chemotaxis of neutrophils. The only known assay for hMDF cannot quantify its level in samples, so an enzyme immunoassay has been standardized for detection of hMDF and hMDF-specific antibodies in test samples. The reported enzyme linked immunosorbent assay (ELISA) was found to be sensitive (89%), specific (91%), accurate (92 2%) and reproducible and was able to detect a minimum concentration of 23 ng hMDF/ml in test samples. The chemotactic factor could be detected in JEV inoculated mouse sera and JEV infected culture fluids. Significant finding of the test was the detection of hMDF in sera of human cases of JE. PMID- 10824199 TI - A four-element based transposon system for allele specific tagging in plants- theoretical considerations. AB - The two-element transposon constructs, utilizing either Ac/Ds or Spm/dSpm, allow random tagging of genes in heterologous model species, but are inadequate for directed tagging of specific alleles of agronomic importance. We propose the use of Ac/Ds in conjunction with Spm/dSpm to develop a four-element system for directed tagging of crop-specific alleles. The four-element based construct would include both Ds and dSpm along with relevant marker genes and would function in two steps. In the first step dSpm(Ds) stocks (a minimum of two) would be crossed to a line containing transposases of Spm and unlinked integrations would be selected from segregating population by the use of a negative selection marker to develop stocks representing integration of dSpm(Ds) at a large number of locations in the genome. Selections would be made for a line in which dSpm(Ds) shows partial or complete linkage to the allele of interest. In the second step selected line would be crossed to a line containing Ac transposase to induce transpositions of Ds element to linked sites thereby exploiting the natural tendency of Ds element to jump to linked sites. Unlinked jumps of dSpm(Ds) and linked jumps of Ds could be monitored by appropriate marker genes. The proposed model would allow tagging of allele of interest in chromosome addition lines and also help in the efficient use of genic male sterility systems for hybrid seed production by tightly marking the fertility restorer gene with a negative selection marker. PMID- 10824200 TI - Degradation and de novo synthesis of D1 protein and psbA transcript levels in Chlamydomonas reinhardtii during UV-B inactivation of photosynthesis and its reactivation. AB - UV-B induces intensity and time dependent inhibition of photosynthetic O2 evolution and PS II electron transport activity in Chlamydomonas reinhardtii. The D1 and D2 proteins of chloroplast membranes are rapidly and specifically degraded in the course of irradiation of cells to UV-B. Continuous synthesis of the two proteins was essential for the repair of damaged PS II as chloramphenicol accelerated UV-B inactivation of photosynthesis and prevented photoreactivation. Northern analysis revealed that UV-B also affected the expression of psbA gene coding for the D1 protein. Cells showing 72% inhibition of PS II activity, revealed a modest net loss of 25% in the level of D1 protein. This shows that synthesis of D1 protein is not the only component involved in the recovery process. Our results indicate that besides affecting the synthesis of the D1 protein UV-B may impair certain post-translational events, which in turn may limit the repair of damaged PS II. PMID- 10824201 TI - Avirulent mutants of Macrophomina phaseolina and Aspergillus fumigatus initiate infection in Phaseolus mungo in the presence of phaseolinone; levamisole gives protection. AB - To evaluate the role of phaseolinone, a phytotoxin produced by Macrophomina phaseolina, in disease initiation, three nontoxigenic avirulent mutants of the fungus were generated by UV-mutagenesis. Two of them were able to initiate infection in germinating Phaseolus mungo seeds only in the presence of phaseolinone. The minimum dose of phaseoli-none required for infection in 30% seedlings was 2 5 mg/ml. A human pathogen, Aspergillus fumigatus was also able to infect germinating seeds of P. mungo in the presence of 5 mg/ml concentration of phaseolinone. Phaseolinone seemed to facilitate infection by A. fumigatus, which is not normally phytopathogenic, by reducing the immunity of germinating seedlings in a nonspecific way. Levamisole, a non-specific immunopotentiator gave protection against infection induced by A. fumigatus at an optimum dose of 50 mg/ml. Sodium malonate prevented the effects of levamisole. PMID- 10824203 TI - Did fleshy fruit pulp evolve as a defence against seed loss rather than as a dispersal mechanism? AB - Relatively few studies have examined the evolution of the mutualism between endozoochorous plants and seed dispersers. Most seed dispersal studies are ecological and examine the role of fruit pulp in promoting seed dispersal. This interaction is often assumed to have originated due to selection stemming from seed dispersers. Here I suggest a "defence scenario" wherein fleshy fruits originated as mechanisms to defend seeds and secondarily became structures to promote seed dispersal. I suggest that frugivory followed from herbivores that specialized on consuming seed defensive tissues and that enhanced seed dispersal was initially a consequence of seed defence. The proposed defence scenario is not posited as an explanation for the sequence that led to all modern frugivores. However, it is suggested that seed predation was the initial source of selection that led to fleshy fruits; the necessary precursor to frugivory. Support is described from the fossil record and from modern structures and interactions. Testable predictions are made in hope that greater interest will be focused on the defensive role of fleshy fruit pulp both in modern interactions and historically. PMID- 10824202 TI - Generation of deviation parameters for amino acid singlets, doublets and triplets from three-dimentional structures of proteins and its implications for secondary structure prediction from amino acid sequences. AB - We present a new method, secondary structure prediction by deviation parameter (SSPDP) for predicting the secondary structure of proteins from amino acid sequence. Deviation parameters (DP) for amino acid singlets, doublets and triplets were computed with respect to secondary structural elements of proteins based on the dictionary of secondary structure prediction (DSSP)-generated secondary structure for 408 selected non-homologous proteins. To the amino acid triplets which are not found in the selected dataset, a DP value of zero is assigned with respect to the secondary structural elements of proteins. The total number of parameters generated is 15,432, in the possible parameters of 25,260. Deviation parameter is complete with respect to amino acid singlets, doublets, and partially complete with respect to amino acid triplets. These generated parameters were used to predict secondary structural elements from amino acid sequence. The secondary structure predicted by our method (SSPDP) was compared with that of single sequence (NNPREDICT) and multiple sequence (PHD) methods. The average value of the percentage of prediction accuracy for a helix by SSPDP, NNPREDICT and PHD methods was found to be 57%, 44% and 69% respectively for the proteins in the selected dataset. For b-strand the prediction accuracy is found to be 69%, 21% and 53% respectively by SSPDP, NNPREDICT and PHD methods. This clearly indicates that the secondary structure prediction by our method is as good as PHD method but much better than NNPREDICT method. PMID- 10824204 TI - Small mammal trapping in tropical montane forests of the upper Nilgiris, southern India: an evaluation of capture-recapture models in estimating abundance. AB - Capture-mark-recapture was used to study small mammal populations in tropical montane forests in southern India. Eleven plots in six montane forest patches were sampled from February-October, 1994. Six species were captured, including four rodents and two shrews. PROGRAM CAPTURE was used to derive estimates of density of the most abundant species in the study area, Rattus rattus Linnaeus. The coefficient of variation of the density estimate was used as an index of precision. The coefficient of variation decreased exponentially with increasing capture probability and with an increase in trapping duration. The coefficient of variation and the capture probability were not correlated with estimates of density. The density estimate increased with trapping duration, as did trap mortality. The latter may have been due to the trend of increased mortality with recaptures of the same individual, which in turn may have been due to weight loss over consecutive captures. Estimates of density derived using four estimators were different for 2, 3, 4 and 5 days of trapping. The coefficient of variation was highest for the generalized removal estimate and lowest for Darroch's estimate. The models and estimators could not be applied to more than one species, and for this species, only in select habitats in a few seasons. Therefore, models of density estimation developed for temperate areas may not be suitable for tropical habitats due to low densities of small mammals in these habitats. PMID- 10824205 TI - Timing of surgery during the menstrual cycle and prognosis of breast cancer. AB - There are conflicting reports on the differential effect of surgery performed during the two phases of the menstrual cycle, namely, follicular and luteal, and prognosis of operable breast cancer. A statistical meta-analysis of the published evidence suggests a modest survival benefit of 15+/-4% when the operation is performed during the luteal phase. Further research in this area might provide a novel avenue to understand the natural history of breast cancer. A spin off from these studies might be the understanding of the importance of events that occur at the time of surgery in determining long term prognosis. PMID- 10824206 TI - The clinical and radiological features of tuberculosis in adolescents. AB - This retrospective review of clinical records and chest radiographs (CR) of adolescents aged 10-18 years was designed to determine age and sex differences in the clinical and radiological features of adolescent tuberculosis (TB). Records of adolescents who were admitted to Brooklyn Hospital for Chest Diseases (BCH) or who were treated at local authority health clinics were screened. Data from 324 adolescents (male:female ratio 1:1.2) were studied. Intra-thoracic lesions were present on CR in 306 (94%). Primary TB with mediastinal adenopathy was present in 32 (10%). Cavitation was present in 180 (56%), 16% at 10 and 73% at 18 years of age. Cavitation occurred in 55% of males and in 56% of females with increasing frequency from 15 years of age in the former and from age 14 in the latter. Microbiological confirmation of diagnosis was obtained in 254 (78%) cases, 52% in those aged 10-13 years and 86% in those > or = 14 years. Pleural effusion was present in 42 (13%), 26 males and 16 females (p < 0.05). Thirteen (7%) of the 182 hospitalized adolescents and 27 (19%) of the ambulant group did not complete therapy. The nature of tuberculous disease in adolescents changed dramatically with increasing age. PMID- 10824207 TI - Chest radiography in ambulatory children with acute lower respiratory infections: effective tuberculosis case-finding? AB - A study was performed to determine the proportion of ambulatory children with acute lower respiratory infections in whom clinical management was changed by findings on routine chest radiography that suggested tuberculosis. The children studied were aged between 2 and 59 months and met the World Health Organization's case definition for pneumonia. They lived in an area with a very high prevalence of tuberculosis. Exclusion criteria included a cough of more than 14 days' duration and a history of a current household contact with active tuberculosis. Twelve (4.4%) of 273 children had radiological findings suggesting tuberculosis, nine of which were suspected mediastinal lymphadenopathy. Eight children were further investigated for tuberculosis: seven of them did not require treatment for tuberculosis and one was lost to follow-up. It is concluded that chest radiography in ambulatory children with acute lower respiratory infections of less than 14 days' duration and not in contact with active tuberculosis does not result in a meaningful increase in the diagnosis of tuberculosis. PMID- 10824208 TI - Group A streptococcal throat colonization is a dynamic phenomenon. AB - In order to gain knowledge about the nature of group A streptococcal throat colonization of asymptomatic children, we studied a cohort of healthy children selected at random. Those with throat cultures positive for group A Streptococci (GAS) were followed for 5 weeks. Repeat throat swabs were obtained at weekly intervals for antigen detection and culture. GAS obtained were serotyped for T antigen. Twenty-two children were identified as having GAS and 50% had a positive antigen test and culture at least five times in 6 weeks. There was no consistent pattern in the typeable organisms. Most individuals had more than one T-type alternating with non-T-typeable organisms. We conclude that colonization may be a dynamic process involving numerous organism serotypes, which wax and wane. PMID- 10824209 TI - Feasibility, acceptability and cost of kangaroo mother care in Recife, Brazil. AB - This descriptive study on kangaroo mother care (KMC) of low-birthweight infants (LBWIs) was carried out in a tertiary care hospital in Recife, Brazil. Of 244 LBWIs weighing less than 1750 g admitted over 14 months, 112 (46%) died before inclusion, 18 (7%) were excluded, and 114 (47%), after stabilization, were cared for by KMC 24 hours a day until discharge. No deaths were recorded in hospital; two twins died of severe pneumonia after discharge and before the age of 3 months. There were no episodes of moderate or severe hypothermia but mild hypothermia (36-36.4 degrees C axillary temperature) occurred at a rate of 30 episodes per 100 infant days, mainly related to occasional separation from the mother. One hundred infants (88%) were discharged on exclusive breastfeeding, eight (7%) were still taking expressed breast-milk from a cup and six (5%) were being fed breast-milk plus formula. The mean daily weight gain during KMC was 15 g. At follow-up, 87% were still exclusively breastfed at 1 month and 63% at 3 months. KMC was acceptable to mothers and staff. An important advantage of KMC over previous conventional care is cost--US$20 vs US$66 per bed/day. This study confirms that KMC for stabilized LBWIs in hospital is feasible, acceptable and cheap and in hospitals with limited resources is an appropriate alternative to conventional incubator care. PMID- 10824210 TI - Alterations in antioxidant status during neonatal sepsis. AB - Septicaemia is a major threat to survival during the early stages of life. There are several reports that suggest that reactive oxygen species (ROs) play a role in a wide variety of diseases. We estimated the activity of xanthine oxidase (XO), malondialdehyde (MDA) content, creatine phosphokinase (CPK) activity, activities of key enzymatic antioxidants, such as superoxide dismutase (SOD), glutathione peroxidase (GPx) and peroxidase (PO), and non-enzymatic antioxidants, viz. uric acid (UA) and albumin (ALB), in 30 neonates with sepsis and 20 age matched controls. The babies were categorized as preterm/term, early onset/late onset, and shock/without shock, as per clinical and laboratory investigations. The study was carried out to evaluate the status of antioxidant enzymes and non enzymatic antioxidants with a view to suggesting the introduction of antioxidant therapy in neonatal sepsis. The activities of serum XO, CPK, SOD and GPx, and the content of MDA were found to be significantly elevated in the neonates with sepsis when compared with controls. Conversely, the activity of PO and the levels of UA and ALB were decreased. The septic, full-term neonates registered significantly higher CPK activity (70%) than the preterm septic neonates. However, infants with late-onset and shock sepsis had a significant decrease in CPK activity (p < 0.05) compared with their corresponding sub-groups. Likewise, UA levels were found to be 28% depressed (p < 0.05) in the babies with late-onset sepsis and 51% increased (p < 0.001) in babies with shock compared with their respective sub-groups. Neonates with septic shock also registered a significant elevation in GPx activity (28%) compared with those without shock. This study suggests increased production of ROs in neonates with sepsis, as evidenced by the positive regulation of XO, SOD and GPx activity. The elevation of antioxidant enzymes, however, was not so effective as to protect from cellular damage and thereby result in higher MDA production. It is evident that antioxidant therapy might be useful in the management of neonates with sepsis but further detailed clinico-biochemical investigations are required to define effective antioxidant therapy. PMID- 10824211 TI - Dietary cyanide from insufficiently processed cassava and growth retardation in children in the Democratic Republic of Congo (formerly Zaire). AB - Dietary cyanide exposure from cyanogenic glucosides in insufficiently processed cassava has been advanced as a contributing factor in child growth retardation. Whether cyanide exposure aggravates children's growth retardation was studied by comparing two populations of children from the northern and the southern zones of the Bandundu region, Democratic Republic of Congo (former Zaire), using dietary interviews, anthropometry and urine analyses. Both populations consumed cassava as their staple diet, but whereas in the north the cassava was well processed, in the south it was inadequately processed. The mean urinary thiocyanate was much higher in the south, whereas mean urinary sulphate excretion was equally low in the two areas. However, the mean urinary SCN/SO4 molar ratio was higher in the south (0.20), indicating that 10-20% of sulphur amino-acids were used for cyanide detoxication. No significant differences were found between the two populations in weight-for-height and weight-for-age indices but the height-for-age index was significantly lower in children from the south, indicating more severe growth retardation in children exposed to dietary cyanide. Because of the preferential use of sulphur amino-acids for cyanide detoxification in the human body, dietary cyanide exposure from cassava may be a factor aggravating growth retardation in Bandundu. PMID- 10824212 TI - Salmonella meningitis in children in Blantyre, Malawi, 1996-1999. AB - Sixty-one episodes of Salmonella meningitis were identified during a 3-year period from February 1996 to January 1999 inclusive. These accounted for 6.8% of all the acute bacterial meningitis cases seen during this time. In contrast, only two children were admitted with Salmonella meningitis in 1982. The increase may reflect the rise in HIV disease and the associated increase in Salmonella septicaemia. All but one child were under 2 years of age, only six children were well nourished and anaemia was common. The prognosis was poor: 33 (58%) died, 19 made a full recovery and five developed sequelae. Two children relapsed, one of whom died. Patients were routinely treated with chloramphenicol, to which all isolates were sensitive in vitro. The poor outcomes suggest that an alternative antibiotic policy is required. PMID- 10824213 TI - Packed red cell volume pattern in Nigerian preterm babies. AB - Weekly packed red cell volume (PCV) was estimated in 57 Nigerian preterm babies whose gestational ages were less than 34 weeks. Babies were excluded if they required exchange blood transfusion or had major congenital malformations. The mean (SD) PCV at birth was 54.6% (8.3). The nadir of the PCV drop was at the 10th 11th week of life. The PCV at the nadir was 29.3% (SD 2.9). The rate of drop in the PCV was most rapid in the 1st 2 weeks (velocity -4.6% per week), but by the 7th week this had decreased to -1.2% per week. The findings indicate a slower rate of postnatal fall in PCV among Nigerian preterms. We recommend that PCV and systemic signs of anaemia be monitored for the 1st 12 weeks of life. PMID- 10824214 TI - The impact of the human immunodeficiency virus type 1 on the management of severe malnutrition in Malawi. AB - A study was undertaken in a central nutritional rehabilitation unit in southern Malawi to assess the impact of HIV infection on clinical presentation and case fatality rate. HIV seroprevalence in 250 severely malnourished children over 1 year of age was 34.4% and overall mortality was 28%. HIV infection was associated significantly more frequently with marasmus (62.2%) than with kwashiorkor (21.7%) (p < 0.0001). Breastfed children presenting with severe malnutrition were significantly more likely to be HIV-seropositive (p < 0.001). Clinical and radiological features were generally not helpful in distinguishing HIV seropositive from HIV-seronegative children. The case fatality rate was significantly higher for HIV-seropositive children (RR 1.6 [95% CI 1.14-2.24]). The increasing difficulties of managing the growing impact of HIV infection on severely malnourished children in Malawi are discussed in the context of reduced support for nutritional rehabilitation units. PMID- 10824215 TI - Neonatal exchange transfusion: a Jordanian experience. AB - The aetiology and complications of exchange transfusion (ET) were studied over a 6-year period in northern Jordan. During that time, 336 neonates (0.46% of total live births) underwent 386 ETs. There was a yearly reduction in the number of ETs, varying from 8.2% in the 1st year of the study to 2.7% in the last year. Thirty-nine (11.9%) required more than one ET. Twenty-five (7.4%) were preterm babies and the remainder full-term. The commonest cause of ET overall was G6PD deficiency, either alone or concomitant with ABO incompatibility (38.1%). ET complications occurred in 51 neonates (15.2%), the commonest being anaemia and bradycardia. Septicaemia occurred in only 3% of cases. Only one baby died. G6PD deficiency, either alone or concomitant with ABO isoimmunization, is the most common cause of ET in northern Jordan. Fewer preterm babies require ET and there is a low incidence of septicaemia following ET. PMID- 10824217 TI - Cystic fibrosis and hypertelorism: a case report. AB - This is a report of cystic fibrosis in association with multiple skeletal defects, including hypertelorism, in a consanguineous family in Sudan. PMID- 10824216 TI - Indian childhood-like cirrhosis in three Saudi Arabian siblings. AB - Three Saudi siblings, two girls and one boy, presented at the ages of 7, 6 and 2.5 years, respectively, and were diagnosed as having features of Indian childhood cirrhosis (ICC). The two girls presented at a late stage of the disease and the boy was diagnosed during routine examination of the family. The initial presenting complaint was abdominal distention and pruritus. All three had a rapid and fatal course. There was no evidence of increased copper ingestion by the families, supporting the suggestion of a hereditary metabolic role in the aetiology of ICC. As far as we are aware, this is the first report of ICC in Saudi Arabian children. PMID- 10824218 TI - Osteomyelitis in sickle cell haemoglobinopathy with elevated fetal haemoglobin. AB - The prevalence rate and characteristics of osteomyelitis in children below 15 years of age were studied in a population characterized by a high prevalence of sickle cell disease (SCD) with elevated fetal haemoglobin (HbF). All children born in our institution routinely undergo a haemoglobinopathy screening test. Osteomyelitis developed in 25 children (16 boys and nine girls) with a mean age of 5.5 years. Fourteen had sickle cell disease (SCD) and 11 had no haemoglobinopathy. The prevalence of osteomyelitis in children with SCD was 69 times higher than in those without (283 vs 4/10(5)). All 14 children with SCD and osteomyelitis also had elevated HbF. Three children had HbS alpha-thalassaemia, with mean HbF 24% and HbS 68.6%. Eleven children had sickle cell anaemia with elevated HbF, mean HbF 31.7% and HbS 66.7%. There was no significant difference in symptoms, signs, leucocyte, neutrophil and band counts, nor in sedimentation rates. Salmonella organisms were significantly more frequently responsible for osteomyelitis in SCD. The presence of elevated HbF in children with SCD should not lead to complacency as they are still at increased risk of developing osteomyelitis. PMID- 10824219 TI - Caffeine: a useful adjunct in the rehabilitation of children suffering from kwashiorkor? PMID- 10824220 TI - In the absence of neonatal screening facilities for sickle cell anaemia. PMID- 10824221 TI - Repeated laboratory investigations and post-procedural radiography: are they warranted in neonatal care? PMID- 10824222 TI - Pragmatic comprehension in secondary school-aged students with specific developmental language disorder. AB - This study explores the hypothesis that there may be particular difficulties for secondary school students with specific developmental language disorder (SDLD) in understanding contextual, pragmatic meaning. Sixty-four SDLD students aged 11+ to 14+ years are compared with chronological-age-matched and language-age-matched non-impaired students. New procedures are used to examine comprehension of two types of ambiguity where the context determines speaker intention: inconsistent messages of emotion and multiple meanings in context. These types of ambiguity are evident in a range of communicative intent, e.g. sarcasm, idiomatic expression, deceit and humour. Preliminary study into adolescent language suggests that at this age there is a growing expectation for students to understand these kinds of communication, both in the classroom and socially. The study finds that the SDLD students were less able than both comparison groups to use context to understand implied meanings. Non-impaired children were also more able to rule out literal interpretations when they did not know the non-literal meaning. These findings were statistically significant. The implications for research and practice are discussed, including those of diagnostic assessment, in the light of the literature survey revealing that many currently available do not assess pragmatic meaning comprehension. There is a challenge to the view that disorders in the semantic and pragmatic domains necessarily co-occur, as suggested by the diagnostic category semantic-pragmatic disorder. PMID- 10824223 TI - Contextual influence on the language production of children with speech/language impairment. AB - To investigate the possible contextual variability of language and interaction, data from 10 children with speech/language impairment and three different types of conversational partners (a parent, a peer and a clinician) were collected. The dialogues were analyzed with respect to the characteristics of the dialogues as whole, to the dominance and the productivity of the three types of conversational partners, and to the productivity, fluency and grammatical structure of the language production of the children with speech/language impairment. Productivity was measured as mean length of utterance (MLU) in words, number of utterances, number of different words and proportion of complete and intelligible utterances. Fluency was measured as the proportion of utterances containing a maze. Grammatical structure was analysed with respect to the use of grammatical morphemes, word order patterns, the occurrence of expansions and the complexity of verb forms. Significant differences, in particular between child-child and adult-child dialogues were found. The adult-child dialogues were asymmetrical, where the adults dominated through asking many questions and talking much. The peer dialogues were more dynamic and equal. In these dialogues the children with speech/language impairment took a more active role as a conversational partner. The children produced more utterances and different words with the adult partners, but variables related to fluency and grammatical structure did not vary as a function of the conversational partner. The results imply that children with speech/language impairment practise different aspects of their communicative ability with different types of conversational partners. In particular, it is important to provide them with opportunities to interact with peers, in order for them to develop their skills as independent conversational partners. PMID- 10824224 TI - Conversational cohesion patterns in men and women with Alzheimer's disease: a longitudinal study. AB - The use of cohesion devices in conversations was examined in 60 individuals (31 women, 29 men) with early to midstage Alzheimer's disease (AD) and 47 non demented elderly (NE) (27 women, 20 men). AD individuals produced more referent errors than NE, although AD and NE subjects otherwise demonstrated similar use of cohesion devices. AD women showed more frequent use of two elliptical devices (clause omission and word omission) compared with AD men and NE men and women. A subset of 23 AD subjects (13 women, 10 men) who completed four data collections (entry, 6, 12 and 18 months) was followed longitudinally. They demonstrated a significant decline in the number of ellipses and conjunctions at 18 months post entry. As AD subjects produced fewer and shorter utterances across time, their use of all cohesive devices declined. No gender differences were found over time. Although referencing errors differentiated early to midstage AD from NE, conversational discourse tasks alone may have limited clinical value to assess and monitor communication competence. PMID- 10824225 TI - Bilingual children in language units: does having 'well-informed' parents make a difference? AB - Findings from a large-cohort study of children with speech and language impairments in language units attached to primary schools across England have suggested that in 11% of the cohort who were bilingual form a subgroup with distinct characteristics. In particular, bilingual children's language difficulties seemed to be more complex and possibly more severe than those of their monolingual peers. It was suggested that these findings might reflect differences in the way that the bilingual children were identified and assessed for speech and language difficulties. Parents of the bilingual children in the original study were interviewed to explore the kind of experiences they had with the identification and assessment process. Differences were found between the bilingual parents and a group of monolingual parents who were also interviewed. Moreover, differences were found between two subgroups of the bilingual parents: those who were 'more informed' and 'less informed' about the process. These differences were found to be related to several other factors, including attitudes to language use within the family and the nature of the parents' relationship with the language unit. PMID- 10824226 TI - Conversation versus narration in pre-school children with language impairment. AB - The study focuses on two elicitation methods for language sampling in children with language impairment: conversation and narration. It has been noted in other studies on different clinical groups that language elicited in different speaking contexts varies in aspects such as MLU, fluency and syntactic complexity. The purpose of this study was to compare genre effects on different aspects of language production in a group of pre-school children with language impairment. The results show that there are differences in language production during conversation compared with narration. Intelligibility and fluency were found to be higher in conversation than in narration, whereas MLU in words was higher in narration. The narrative task elicited more phrasal expansions and grammatical morphemes per utterance than the conversation. However, the children used more complex verb forms in conversation than in narration. The results are discussed in relation to recent research. PMID- 10824227 TI - Limitations in working memory: implications for language development. AB - In this study, the proposal that individual differences in spoken language acquisition may be due to limitations in short-term memory abilities was investigated within a working memory framework. The relationship speech production skills and working memory abilities was examined in two groups of 4 year-old children, matched for non-verbal ability but who had either relatively good or poor non-word repetition skills. Children with better non-word repetition skills produced speech that comprised a wider repertoire of words, on average longer utterances and a greater range of syntactic constructions than did children with relatively poor non-word repetition skills. The significant association found between these indices of language development and verbal short term memory span assessed with non-spoken recall, suggested that this relationship was not merely due to the common output requirements of the language and memory tasks. Inconsistent associations between language performance and two tasks of visuo-spatial short-term memory precluded firm conclusions being drawn regarding the specificity of the relationship to the phonological domain. Cognitive mechanisms that may underlie the association between spoken language development and working memory skills are discussed. PMID- 10824228 TI - Relationships between symbolic play, functional play, verbal and non-verbal ability in young children. AB - It is well established that certain aspects of play in young children are related to their emerging linguistic skills. The present study examined the relationships between functional play, symbolic play, non-verbal ability, and expressive and receptive language in normally developing children aged between 1 and 6 years using standardized assessment procedures, including a recently developed Test of Pretend Play (ToPP). When effects of chronological age were partialled out, symbolic play remained significantly correlated with both expressive and receptive language, but not with functional play or non-verbal ability; and functional play was only correlated significantly with expressive language. It is concluded that ToPP will provide practitioners with a useful way of assessing symbolic ability in children between the ages of 1 and 6 years, and will contribute to the assessment and diagnosis of a number of communication difficulties, and have implications for intervention. PMID- 10824229 TI - Pitch change in male-to-female transsexuals: has phonosurgery a role to play? AB - Male-to-female transsexuals, who have undergone gender reassignment surgery, may continue to have low pitched voices. Voice therapy may assist them to use their voice in a manner more likely to be perceived as female but, if this approach is unsuccessful, a laryngeal framework operation may be necessary to raise vocal pitch. This study assessed the effects of crico-thyroid approximation surgery in 14 transsexuals. Modal pitch was significantly increased by surgery but with substantial variation across speakers. Modal pitch was significantly correlated with judgements of gender by speech and language therapists who listened to tapes of the subjects. The results suggest that crico-thyroid approximation may be used to raise the pitch of voices of male-to-female transsexuals. Further research is needed to clarify the reasons for the variability in outcome, to monitor the longer-term changes in voice and the impact for clients of their modified voice in real life situations. PMID- 10824230 TI - An analysis of referrals to speech and language therapy in 11 centres, 1987-95. AB - This retrospective study analyses data collected on an information system developed specifically for the use of speech and language therapists in the management of their services. Eleven healthcare providers have used the system since 1985. This paper reflects data on 73,000 clients referred to these speech and language therapy services between 1987 and 1995. The annual referral rate to these 11 trusts in 1987 was 4129, rising to 11,944 in 1995. The percentage referrals per head of population per year varies between 0.32 and 0.58% of the population. Of referrals to speech and language therapy, 96% comes from 24 different sources, including self referrals and referrals from other agencies, such as education. The top five referrers in 1987 were health visitors, educational service, general medicine/geriatrics, general practitioners (GPs), and other speech and language therapists. In 1995 ear, nose and throat (ENT) services replaced speech and language therapists in the top five referrers. PMID- 10824231 TI - Staff communication with people with intellectual disabilities: the impact of a work-based training programme. AB - Previous studies have identified changes that front-line service staff could usefully make to enhance their communications and those of their clients who have intellectual disabilities. These were incorporated into a training programme delivered in the workplace that involved a self-selected group of 24 experienced staff working either in small-scale residential settings and day centres. Analysis of video-recordings made before and after training showed that although most clients had become more active communicators (particularly when they were engaged in shared activities with staff), significant changes in staff behaviours as a whole were not observed. However increased responsiveness from staff did correlate significantly with increases in the client's communication acts. In addition, qualitative reports from staff and tutors pinpointed specific changes that staff had made. The difficulties of evaluating changes in staff-client communications are discussed and four features for training staff in communication are identified as essential; namely, it should be work-based, client focused, mentor-guided, and effective strategies documented and shared. PMID- 10824232 TI - Congenital malformations of the ear and cochlear implantation in children: review and temporal bone report of common cavity. AB - The objective of this review is to analyze aspects of congenital malformation of the ear in relation to cochlear implantation in children. Having briefly described the in utero development of the ear and the classification of types of external, middle and inner ear malformation, five practical aspects of these malformations are discussed. It seems likely that the combination of bilateral profound sensorineural deafness with bilateral microtia severe enough to make a surgical approach to the cochlea difficult will be extremely uncommon. No such cases have been reported, although Klippel-Feil deformity seems the syndrome most likely to produce this set of circumstances. Abnormalities in the intratympanic course of the facial nerve have been associated with cochlear malformation, emphasizing the benefit of intra-operative facial nerve monitoring, and a technique suggested for safely avoiding an abnormally placed nerve. Fistulae of cerebrospinal fluid (CSF) and perilymph can complicate surgery and are relatively common in common cavity and Mondini malformations. Strategies for facilitating surgery in the presence of 'gushers', for measuring the pressure of a gusher and for placement of the cochlear implant electrode array are reviewed, with reports of fluctuating levels of electric current when implants lie in dysplastic cochleas. The relationship of implant performance to VIIIth nerve tissue in malformed cochleas is discussed, with a description of the histological findings in a common cavity cochlea. Techniques for identifying the absence of the cochlear nerve are reviewed. Stimulation of the facial nerve by cochlear implants has been described in cases of congenital malformation of the labyrinth but is relatively uncommon. Case reports of the benefit received by implanted children with congenital cochlear malformation have appeared since 1988. Most cases reported have not yet been followed for long enough to establish a clear picture of the outcome following cochlear implantation in such children; no centre has yet built up a large series of cases, but there have been two multicentre postal surveys. It seems likely that in cochlear malformation the range of potential outcomes in terms of hearing threshold and the development of speech perception and production will be similar to the range found in implanted children without cochlear dysplasia. However there is, as yet, no clear picture of the mean level of performance within this range. PMID- 10824233 TI - Do we need to change the WHO definition of osteoporosis? PMID- 10824234 TI - An update on the diagnosis and assessment of osteoporosis with densitometry. Committee of Scientific Advisors, International Osteoporosis Foundation. AB - In 1994 the WHO proposed guidelines for the diagnosis of osteoporosis based on measurement of bone mineral density. They have been widely used for epidemiological studies, clinical research and for treatment strategies. Despite the widespread acceptance of the diagnostic criteria, several problems remain with their use. Uncertainties concern the optimal site for assessment, thresholds for men and diagnostic inaccuracies at different sites. In addition, the development of many new technologies to assess the amount or quality of bone poses problems in placing these new tools within a diagnostic and assessment setting. This review considers the recent literature that has highlighted the strengths and weaknesses of diagnostic thresholds and their use in the assessment of fracture risk, and makes recommendations for actions to resolve these difficulties. PMID- 10824235 TI - Hand ultrasound for osteoporosis screening in postmenopausal women. AB - There is a need for low-cost screening methods to detect low bone mass (osteopenia or osteoporosis) in postmenopausal women. The utility of quantitative ultrasonography (QUS) of the hand was assessed for osteoporosis screening using the WHO criteria. Bone mineral density (BMD) was measured in 206 postmenopausal Mexican-American women at the total hip and lumbar spine by dual-energy X-ray absorptiometry (DXA). The amplitude-dependent speed of sound (AD-SoS) was measured in the phalanges by QUS. Subjects identified by DXA as having osteopenia or osteoporosis had significantly lower AD-SoS values in comparison with normals. Estrogen users had significantly higher spine and hip BMD and AD-SoS values compared with non-estrogen users. The areas under the receiver operating characteristic (ROC) curves (AUC) for AD-SoS to screen for osteoporosis (T-score < or = -2.5) at the spine or hip were 0.73 for all subjects, 0.74 for estrogen users and 0.68 for non-estrogen users. The AUC for non-estrogen users to screen for osteopenia (T-score -1 to -2.5) was 0.77. Performance comparisons of AD-SoS with SCORE (a risk factor questionnaire) and body weight showed AUC values of 0.73, 0.69 and 0.65, respectively. QUS was the superior screening test when considering both the AUC and the shape of the ROC curves. For non-estrogen users, the group at higher risk for osteoporosis, QUS correctly identified 31% as normal, and 62% as having low bone mass and needing DXA referral; and the remaining 7% were false negatives. These data suggest phalangeal QUS can be effectively used for screening osteoporosis in postmenopausal women. PMID- 10824236 TI - Identification of early postmenopausal women with no bone response to HRT: results of a five-year clinical trial. AB - Hormone replacement therapy (HRT) prevents postmenopausal bone loss and fractures. However, the occurrence of women with no bone response to HRT has not been widely examined. We identified the densitometric nonresponders to long-term HRT and investigated some characteristics and biochemical variables as possible predictors of densitometric nonresponse in postmenopausal women. The study population was a subsample of the Kuopio Osteoporosis Study (n = 14,220). A total of 464 early postmenopausal women were randomized into four treatment groups: (1) HRT (sequential combination of 2 mg estradiol valerate and 1 mg cyproterone acetate); (2) vitamin D3; (3) HRT + Vitamin D3 combined; and (4) placebo. In this study, the data from HRT and placebo groups were analyzed. Lumbar (L2-4) and femoral neck bone mineral density (BMD) were determined by dual-energy X-ray absorptiometry (DXA) at baseline and after 5 years of treatment. A densitometric nonresponder was defined as a woman whose 5-year BMD change was similar to the mean BMD change (+95% CI) of the placebo group or worse. Altogether, 74 women in the HRT group and 104 women in the placebo group complied with the treatment. According to spinal BMD analysis, 11% of the women were classified as densitometric nonresponders; the corresponding proportion for femoral BMD analysis was 26%. Both smoking (p = 0.003) and low body weight (p = 0.028) were significant risk factors for densitometric nonresponse to HRT. After 6 months of treatment the densitometric nonresponders (hip) had a significantly higher mean serum follicle stimulating hormone (FSH) level (p = 0.038) and lower increases in serum estradiol levels (p = 0.006) than the densitometric responders. The mean changes in serum FSH and alkaline phosphatase levels were significantly lower among the densitometric nonresponders (spine) than responders (p = 0.043 and 0.017, respectively). In conclusion, this prospective study shows that especially current smokers and women with low body weight are at increased risk of poor bone response to HRT. Repeated serum FSH, estradiol and alkaline phosphatase measurements during the first months of long-term HRT may be helpful in identifying the women with no bone response to HRT. PMID- 10824237 TI - Risk factors for perimenopausal fractures: a prospective study. AB - This prospective study was aimed at determining the risk factors for the development of fractures in perimenopausal women. The study group (n = 3068) was comprised of a stratified population sample of women aged between 47 and 56 years. During the follow-up period of 3.6 years, 257 (8.4%) of the women sustained a total of 295 fractures. After adjustment for covariates, the relative risk (RR) of sustaining a fracture was found to be 1.4 [95% confidence interval (CI) 1.2-1.6] for a 1 standard deviation (SD) decrease in the spinal and femoral neck bone mineral density (BMD). Women with a previous fracture history were found to have an increased risk of fracture [RR 1.7 (95% CI 1.3-2.2)] and those reporting three or more chronic illnesses exhibited a RR of 1.4 (95% CI 1.0-1.9). Women not using hormone replacement therapy (HRT) had a RR of 1.5 (95% CI 1.1 2.2) for all fracture types. When osteoporotic fractures (vertebral, hip, proximal humerus and wrist fractures; n = 98) were used as an endpoint, the independent risk factors were found to be a low BMD (RR for a 1 SD decrease in both spinal and femoral neck BMD was 1.6, 95% CI 1.3-2.0), a previous fracture history (RR 1.9, 95% CI 1.3-2.9) and nonuse of HRT (RR 2.2, 95% CI 1.3-4.0). The independent risk factors for all other fractures (n = 158) were a low BMD (RR for a 1 SD decrease in the spinal BMD was 1.4, 95% CI 1.2-1.6 and in the femoral neck BMD was 1.3, 95% CI 1.1-1.5), a previous fracture history (RR 1.6, 95% CI 1.1 2.2), smoking (RR 1.8, 95% CI 1.1-2.7) and having had three or more chronic illnesses (RR 1.6, 95% CI 1.1-2.2). Weight, height, age, menopausal status, maternal hip fracture, use of alcohol, coffee consumption or dietary calcium intake were not independently associated with the development of any particular type of fracture. We conclude that the independent risk factors for perimenopausal fractures are a low bone density, previous fracture history, nonuse of HRT, having had three or more chronic illnesses and smoking, the gradient of risk being similar for spinal and femoral neck BMD measurements in the perimenopausal population. The risk factors are slightly different for perimenopausal osteoporotic than for other types of fractures. PMID- 10824238 TI - Factors associated with mortality after hip fracture. AB - There is a well-known excess mortality subsequent to hip fracture, which is probably restricted to subgroups of hip fracture patients with reduced health status. We studied the association between risk factors and death in 248 hip fracture patients and 248 controls originally enrolled in a population-based case control study. This cohort was followed for 3 1/2 years with respect to total mortality. A markedly increased mortality was found in hip fracture patients passing a mental status test at a low score [relative risk (RR) = 2.3, 95% confidence interval (CI) 1.4-3.7], in hip fracture patients reporting two or more selected chronic diseases (RR = 3.3, 95% CI 1.8-6.1), in hip fracture patients not walking outdoors before the fracture (RR = 3.2, 95% CI 2.0-5.1) and in hip fracture patients in the lower half of handgrip strength distribution (RR = 2.3, 95% CI 1.6-3.4), all compared with the control group. In contrast, hip fracture patients without these risk factors did not have increased mortality compared with the control group. This study suggests that otherwise healthy and fit patients do not have increased mortality subsequent to hip fracture. The excess mortality is restricted to persons with reduced mental status, reduced somatic health and low physical ability. Special attention should be paid to patients with such risk factors in the treatment and rehabilitation period. PMID- 10824239 TI - Screening for osteoporosis using easily obtainable biometrical data: diagnostic accuracy of measured, self-reported and recalled BMI, and related costs of bone mineral density measurements. AB - The aims of the present study were: to determine the diagnostic accuracy of objectively measured, self-reported and recalled body mass index (BMI) for osteoporosis and osteopenia; to determine the diagnostic costs, in terms of bone mineral density (BMD) measurements, per osteoporotic or osteopenic patient detected, using different BMI tests; and to determine the extent to which the results can be used within the framework of the current screening program for breast cancer in The Netherlands. Within the framework of a cross-sectional study on the prevalence of osteoporosis in the south of The Netherlands, 1155 postmenopausal women aged 50-80 years were asked for their present height and their weight at age 20-30 years. Subsequently their actual weight, height and BMD of the lumbar spine (DXA) were measured. The BMD cutoff was 0.800 g/cm2 for osteoporosis and 0.970 g/cm2 for low BMD (osteoporosis + osteopenia). After receiver operating characteristic analysis, age was cut off at 60 years and BMI at 27 kg/m2. Diagnostic accuracies of objectively measured, self-reported and recalled BMI were evaluated using predictive values (PV) and odds ratios. The resulting 'true positive' and 'false positive' rates were used to calculate diagnostic costs (i.e., DXA) for each osteoporotic patient or low-BMD patient detected. The prevalence of osteoporosis in the study population was 25%, that of low BMD 65%. Only the age-BMI tests 'age > or = 60, BMI < or = 27' showed PVs for osteoporosis (31-41%) and for low BMD (71-81%) that were higher than the prior probabilities for these conditions. Related odds ratios were 2.14-3.18 (osteoporosis) and 1.87-3.04 (low BMD). The objective BMI test detected 50% of the osteoporotic patients. Using the self-reported BMI test and the recalled BMI test, detection rates increased to 55% and 69%, respectively. Concomitant costs per osteoporotic patient detected rose by 24%. Detection of patients with a low BMD increased from 38% for objective BMI and 42% for self-reported BMI to 60% for recalled BMI. Related costs increased by 11%. If all women over 50 years of age (irrespective of their BMI) were to be referred for BMD measurement, costs per osteoporotic patient or low-BMD patient detected would be 304 and 116 Euros, respectively. Only in women over 60 years does a BMI below 27 kg/m2 provide a better prediction of the presence of osteoporosis or low BMD than could be expected solely on the basis of the relevant prevalences in postmenopausal women aged 50-80 years. If the use of BMI for the detection of osteoporotic or low-BMD patients is still considered, measuring weight and just asking for a person's height will do. Although age and BMI are the strongest risk factors for osteoporosis, they are of less significance when used for screening the population for osteoporosis. More research is needed before age and BMI can be included in any screening program. As regards practical considerations alone, measurements of BMD could be implemented within the screening program for breast cancer. PMID- 10824240 TI - Radiographic absorptiometry of the middle phalanx (digit II) in a Caucasian pediatric population: normative data. AB - The availability of improved therapies for children with diseases affecting bone growth and/or metabolism has caused increased interest in bone mineral density (BMD) assessment. The purpose of this study was to determine normal values for phalangeal radiographic absorptiometry (RA) in a Caucasian pediatric population. Five hundred and seventy-two healthy Caucasian children and adolescents (aged 5 19 years) were enrolled in this study. For RA one posteroanterior exposure of the left hand and one lateral exposure of the left index finger were taken. All films were analyzed, yielding BMD (mg Al/mm3) values for two phalangeal sites: one at the proximal quarter of the phalangeal length (BMD25%) and the second at the midpoint of the phalangeal length (BMD50%). Also skeletal age (SA) was assessed, as data normalized for SA can be used in populations which show a dissociation between SA and calendar age. We found that the BMD25% was significantly higher in girls than in boys for the SA group 11-14 years. The BMD50% was significantly higher in girls than in boys for the SA group 11-15 years. Our data show that BMD25% remains fairly constant until the SA of 12.0 years in boys and 10.1 years in girls; after these skeletal ages BMD shows a sharp increase. The same applies to BMD50%, which remains fairly constant until the SA of 12.4 years in boys and 10.7 years in girls. In this paper we present normative data for RA in a pediatric population. These data normalized for skeletal age could be implemented in a clinical setting. PMID- 10824241 TI - Validity of self-report of fractures: results from a prospective study in men and women across Europe. EPOS Study Group. European Prospective Osteoporosis Study Group. AB - In population-based studies of osteoporosis, ascertainment of fractures is typically based on self-report, with subsequent verification by medical records. The aim of this analysis was to assess the validity of self-report of incident nonspine fractures using a postal questionnaire. The degree of overreporting of fracture (false positives) was assessed by comparing self-reports of new fracture from respondents in the multicenter European Prospective Osteoporosis Study with data from other sources including radiographs and medical records. In the analysis, 563 subjects reported nonspine fractures. Verification of the presence of fracture was possible in 510 subjects. Of these, fractures were not confirmed in 11% (false positives). The percentage of false positives was greater in men than in women (15% vs 9%, p = 0.04), and less for fractures of the distal forearm and hip than for fractures at other sites. In a separate study, the degree of underreporting (false negatives) was assessed by follow-up of 251 individuals with confirmed fracture ascertained from the records of fracture clinics in three European centers (Lubeck, Oviedo, Warsaw). Questionnaire responses were received from 174 (69%) subjects. Of these, 12 (7%) did not recall sustaining a fracture (false negatives). The percentage of false negatives was lower for hip and distal forearm fractures with only 3 of 90 (3%) such fractures not recalled. Using the combined data from both studies, of those who reported a 'date' of fracture on the questionnaire, 91% of subjects were correct to within 1 month of the actual date of the fracture. A postal questionnaire is a relatively simple and accurate method for obtaining information about the occurrence of hip and distal forearm fractures, including their timing. Accuracy of ascertainment of fractures at other sites is less good and where possible self-reported fractures at such sites should be verified from other sources. PMID- 10824242 TI - Quantitative ultrasound and bone mineral density in patients with primary hyperparathyroidism before and after surgical treatment. AB - The aim of this study was to assess the pattern of ultrasound (QUS) parameters and bone mineral density at different skeletal sites in patients with primary hyperparathyroidism (PHPT) before and after surgical treatment. In 22 patients (age range 28-74 years) with PHPT we measured speed of sound (SOS), attenuation (BUA) and Stiffness at the calcaneus, amplitude-dependent speed of sound (AD-SoS) at proximal phalanges, and bone mineral density at lumbar spine (BMD-LS) and at the mid-radius (BMD-MR) and ultra-distal radius (BMD-UDR) before, 1 and 2 years after surgical operation. Twenty-two age- and sex-matched healthy subjects provided control data. Before surgery, all parameters apart from SOS were significantly lower in PHPT patients than in controls. At the end of the study period, BMD-LS increased by 7.0%, BMD-UDR by 7.4% and BMD-MR by 11.0%. The changes in ultrasound parameters after surgery were lower (0.44% for SOS, 2.2% for BUA, 3.3% for Stiffness and 2.6% for AD-SoS); however, the increase was statistically significant (p < 0.05 and p < 0.01, respectively) only for Stiffness and AD-SoS. Our results indicate that parathyroidectomy increases both axial and appendicular BMD and influences QUS parameters differently at the calcaneus and at the phalanges. The combined use of BMD and QUS could improve the assessment of skeletal status in patients with PHPT before and after surgery. PMID- 10824243 TI - Bone loss following tibial osteotomy: a model for evaluating post-traumatic osteopenia. AB - The reduced bone mineral density (BMD) found in patients with fractures may, in part, follow rather than precede the fracture. We studied the magnitude and reversibility of bone loss in the 15 months following osteotomy in 21 men and 5 women with localized medial arthritis of the knee. BMD (mean +/- SD), measured using dual-energy X-ray absorptiometry, decreased by a maximum of 35 +/- 21% in the mid-diaphysis of the affected tibia at 9 months after surgery (p < 0.001). At 15 months, reversal of bone loss in non-fractured bones was incomplete; the remaining deficit was 20 +/- 27% relative to baseline (p < 0.001). Maximum bone loss occurred at 9 months at the total body (5 +/- 2%), spine (15 +/- 17%) and at Ward's triangle of the proximal femur of the unoperated limb (10 +/- 17%) (all p < 0.01). In summary, post-traumatic bone loss is region-specific with incomplete reversibility, at least after about 15 months. Deficits in BMD in cross-sectional studies of patients with fractures, held to be responsible for the bone fragility, may, in part, follow rather than precede the fracture. PMID- 10824244 TI - Risk factors for perimenopausal distal forearm fracture. AB - This prospective population-based cohort study investigated factors predicting distal forearm fracture (DFF) in perimenopausal women. The study population consisted of 11,798 women from the Kuopio Osteoporosis Risk Factor and Prevention (OSTPRE) Study in Finland. Mean baseline age of these women was 52.3 (SD 2.9) years (range 47-56 years) and 68% were postmenopausal. Three hundred and sixty eight women (3.1%) had a validated DFF during the 5-year follow-up. Previous wrist fracture, postmenopausal state, age and nulliparity were independent predictors of DFF, while hormone replacement therapy (HRT), dairy calcium and overweight protected against it in multivariate Cox regression analysis: previous wrist fracture increased the DFF risk by 158% (p < 0.0001), menopause by 69% (p = 0.002) and age by 6% per year (p = 0.010), whereas the continuous use of HRT decreased the risk by 63% (p = 0.0001), the use of dairy calcium at 1000-1499 mg/day (vs < 500 mg/day) by 39% (p = 0.004), overweight (BMI > 25 kg/m2) by 36% (p = 0.0002) and parity by 29% (p = 0.031). Combining dichotomous low weight, low use of calcium, non-use of HRT and previous wrist fracture into a risk score gave a dose-response effect by score level: the presence (vs absence) of all four risk factors resulted in a 12-fold DFF risk. Nevertheless, the sensitivity and specificity of the score for detecting DFF remained low. It was concluded that HRT, high nutritional calcium intake and overweight protect against but a history of wrist fracture predisposes to perimenopausal distal forearm fracture. A simple risk factor inquiry would help to identify perimenopausal women at high risk of distal forearm fracture. PMID- 10824246 TI - Biochemical variables in pre- and postmenopausal women: reconciling the calcium and estrogen hypotheses. PMID- 10824245 TI - Biochemical responses of bone metabolism to 1,25-dihydroxyvitamin D administration in black and white women. AB - The basis for the racial difference in bone mass between black and white women is not known. Lower bone turnover, better renal calcium conservation, and decreased sensitivity to parathyroid hormone (PTH) have been proposed as explanations. A dynamic comparison of osteoblast function, utilizing stimulation by 1,25 dihydroxyvitamin D [1,25(OH)2D], has not been tested between these two ethnic groups. We compared well-matched black (n = 15) and white (n = 15) premenopausal women, before and during 5 days of 1,25(OH)2D administration (1.0 microgram/day) in order to assess dynamic indices of bone metabolism. As expected, at baseline, black women had lower levels of serum 25-hydroxyvitamin D and biochemical markers of bone turnover with slightly higher levels of PTH. Black women also had superior renal calcium conservation than white women at baseline. In response to 1,25(OH)2D administration, black women had a slightly greater increase in serum calcium and greater decrement in PTH. Moreover, black women showed a lesser increment in urinary calcium than white women and a more robust increase in two markers of bone formation--osteocalcin and carboxyterminal propeptide of type 1 procollagen--than white women. There were no changes in bone resorption indices in either race upon 1,25(OH)2D administration. These data provide preliminary evidence that black women conserve calcium more efficiently under both static and dynamic conditions, and also appear to have better osteoblastic functional reserve than white women. PMID- 10824247 TI - Mirror image--the care of the professional voice. PMID- 10824249 TI - Some medical aspects of a round the world yacht race. PMID- 10824248 TI - Jet lag/night shift, blindness and melatonin. PMID- 10824250 TI - Medical approaches to snoring and sleep apnoea. PMID- 10824251 TI - "How do snorers snore"? PMID- 10824252 TI - Lasers--the non surgical approach to ablating diseased tissue. PMID- 10824253 TI - Medicine facing modernity. PMID- 10824254 TI - "Post-traumatic stress--friend or foe"? PMID- 10824255 TI - Opium lecture for Medical Society of London. PMID- 10824256 TI - Drug cultures in context. PMID- 10824257 TI - Is an athlete born or made? PMID- 10824258 TI - Lettsomian lecture. Helicobacter pylori--the germ at the bottom of your stomach. PMID- 10824259 TI - Can we believe the experts? PMID- 10824260 TI - The annual oration. 1773. PMID- 10824261 TI - Responses to light after retinal degeneration. AB - Transgenic rodless mice were given 1-h pulses of light of varying brightness at times of the night when they were normally active. The rodless mice showed decreases in locomotor activity during light pulses brighter than 2 lux; these decreases were significantly greater than those in wildtypes (ANOVA, P < 0.01). However, with very dim light, rodless mice showed no changes in activity, whereas wildtype mice actually increased their activity. It is suggested that irradiance detection could be enhanced by absence of image-forming vision. Enhanced inhibition of activity around twilight may be adaptive for mice in some circumstances and so help maintain genes for retinal degeneration in natural populations. PMID- 10824262 TI - Contribution to the kinetics and amplitude of the electroretinogram b-wave by third-order retinal neurons in the rabbit retina. AB - The ERG b-wave is widely believed to reflect mainly light-induced activity of on center bipolar cells and Muller cells. Third-order retinal neurons are thought to contribute negligibly to generation of the b-wave. Here we show that pharmacological agents which affect predominantly third-order neurons alter significantly both the kinetics and amplitude of the b-wave. Our results support the notion that changes in the amplitude and kinetics of light-induced membrane depolarization in third-order neurons produce similar changes in the amplitude and kinetics of the b-wave. We conclude that activity in third-order neurons makes a significant contribution to b-wave generation. Our results also provide evidence that spiking activity of third-order neurons truncates the a-wave by accelerating the onset of the b-wave. PMID- 10824263 TI - Spectral sensitivity of photoreceptors in an Australian marsupial, the tammar wallaby (Macropus eugenii). AB - Microspectrophotometric measurements on the rod photoreceptors of the tammar wallaby showed that they have a peak absorbance at 501 nm. This indicates that macropod marsupials have a typical mammalian rhodopsin. An electroretinogram based study of the photoreceptors confirmed this measurement and provided clear evidence for a single middle wavelength-sensitive cone pigment with a peak sensitivity at 539 nm. The electroretinogram did not reveal the presence of a short-wavelength-sensitive cone pigment as was expected from behavioural and anatomical data. Limitations of the electroretinogram in demonstrating the presence of photopigments are discussed in relation to similarly inconsistent results from other species. PMID- 10824264 TI - Coarse-grained information dominates fine-grained information in judgments of time-to-contact from retinal flow. AB - To investigate the relative importance of fine- and coarse-grained structure in the analysis of retinal flow, subjects made estimates of time-to-contact from random dot kinematograms depicting movement towards a flat, sparsely textured surface. Individual display elements moved smoothly away from each other while expanding smoothly in size. By artificially manipulating the rate at which the individual elements expanded we showed that this cue has only a small effect upon performance. When individual elements were replaced by small clusters of dots, expansion of the clusters had a similarly small effect upon performance. However, estimates of time-to-contact were possible when a single expanding cluster was presented in isolation. We conclude that both types of information are available to the subject but that estimates of time-to-contact are based primarily on coarse-grained changes in the position of image elements and that fine-grained changes in element size or position play only a minor role. PMID- 10824265 TI - What covariance mechanisms underlie green/red equiluminance, luminance contrast sensitivity and chromatic (green/red) contrast sensitivity? AB - In order to investigate the mechanisms underlying green/red equiluminance matches in human observers and their relationship to mechanisms subserving luminance and/or chromatic (green/red) contrast sensitivity, we tested 21 human subjects along these dimensions at 16 different spatial and temporal frequencies (spatial frequency, 0.25-2 c/deg; temporal frequency, 2-16 Hz) and applied factor analysis to extract mechanisms underlying the data set. The results from our factor analysis revealed separate sources of variability for green/red equiluminance, luminance sensitivity and chromatic sensitivity, thus suggesting separate mechanisms underlying each of the three main conditions. When factor analysis was applied separately to green/red equiluminance data, two temporally-tuned factors were revealed (factor 1, 2-4 Hz; factor 2, 8-16 Hz), suggesting the existence of separate mechanisms underlying equiluminance settings at low versus high temporal frequencies. In addition, although the three main conditions remained separate in our factor analysis of the entire data set, our correlation matrix nonetheless revealed systematic correlations between equiluminance settings and luminance sensitivity at high temporal frequencies, and between equiluminance settings and chromatic sensitivity at low temporal frequencies. Taken together, these data suggest that the high temporal frequency factor underlying green/red equiluminance is governed predominantly by luminance mechanisms, while the low temporal frequency factor receives contribution from chromatic mechanisms. PMID- 10824266 TI - Textured backgrounds alter perceived speed. AB - Both the luminance contrast of an object, and the nature of the background texture over which it moves, are known to influence its perceived speed. In this study the effect of object contrast upon perceived speed was investigated for targets moving across textured patterns of various contrasts. Experiment 1 showed a strong effect of contrast for objects moving over homogenous backgrounds, that was reduced or abolished if the object moved over a textured background. A further experiment suggested that this reduction may be the result of an increase in target visibility, perhaps as a result of additional 'second order' motion signals produced by motion over texture backgrounds. A final experiment suggested that two processes were occurring: (1) higher contrast backgrounds appeared to increase the perceived speeds of all objects; and (2) that higher contrast backgrounds eliminated the contrast induced changes in perceived speed. PMID- 10824267 TI - Contour integration in color vision: a common process for the blue-yellow, red green and luminance mechanisms? AB - We compare the performance of the red-green, blue-yellow and luminance postreceptoral mechanisms on a contour integration task requiring the linking of oriented Gabor elements across space to extract a winding 'path' or contour. We first establish that for all three mechanisms curvature and contrast are independent; losses in performance due to one cannot be compensated by changes in the other. We then compare contour integration by the three mechanisms using a method that controls for their differences in cone contrast thresholds. Our results show that despite the poor orientation discrimination thresholds and poor spatial sampling found for the blue-yellow mechanism, all three mechanisms perform similarly on contour integration over a wide range of curvatures. Furthermore, all three mechanisms have the same dependence on path curvature. We also investigate the effects of adding external orientation noise. Our results imply that the internal orientation noise for extracting 'aligned' path elements is similar in the three mechanisms and for all path curvatures, and the relative efficiencies are also similar for the three mechanisms. To account for our results, we propose that the three postreceptoral mechanisms use a common contour integration process. This linking process, however, cannot be color-blind; our last experiment shows that linking between different chromatic mechanisms or between opposite spatial phases disrupts contour integration. We thus propose that the common integration process remains sensitive to the color contrast and phase of its inputs. PMID- 10824268 TI - Luminance processing in apparent motion, Vernier offset and stereoscopic depth. AB - We obtained (apparently) linear responses to luminance from three special displays of apparent motion, Vernier offset and stereoscopic depth. In our motion stimulus a dark and a light bar exchanged luminances repetitively on a grey surround. Motion was attributed to the bar that differed more from the surround, that is, on a dark surround the light bar appeared to jump, and on a light surround the dark bar appeared to jump. The apparent motion disappeared when the luminance of the surround lay halfway between that of the bars--on a linear, not a logarithmic scale. Similar results were obtained for special Vernier offset and stereo stimuli. These results cannot be explained if all luminances are processed within the same luminance pathway and that pathway transforms input luminance using non-linear compression. However, the apparent linearity of our results could arise from opposite and equal non-linearities cancelling out within separate ON- and OFF-spatial luminance pathways. A second set of experiments presented one bar separately into each eye on different surrounds (dichoptic presentation of competing apparent motion signals) or manipulated the display spatially so that different surrounds were associated with different bars (binocular presentation of competing Vernier targets). Results showed that apparent motion and Vernier signals of equal Weber contrast (normalisation of linear difference to surround luminance) evoked equal-motion and equal Vernier offset strengths. Given that motion and Vernier strength followed Weber's law, we infer that the ON- and OFF-pathways transform luminance non-linearly. Our third experiment presents an example of a brightness bisection task in which we were able to influence the bisection steps, to follow either a linear or non-linear series. The benefits of parsing the visual scene so that visual information is processed within two opposite luminance pathways is discussed. PMID- 10824269 TI - The spatial arrangement of L and M cones in the peripheral human retina. AB - The spatial arrangement of L and M cones in the human peripheral retina was estimated from red-green color naming of small test flashes (0.86 min of arc, 555 nm, constant intensity) presented at different locations (grid with 1.5 min of arc steps) centered at 17 degrees temporal eccentricity. Simulated red-green color naming ratings were generated by a model based on an ideal observer for all possible patterns of placement and relative numerosities of L and M cones, constrained by the anatomical data on the statistics of cone spacing at this retinal location. The best matching simulated performance as compared to the human observer's data determined the cone array most likely to produce that observer's color naming results. The mosaics for two color normal observers showed L and M cones randomly arrayed over this retinal region. Consequences of random cone placements for spectral sampling and color opponency are discussed. PMID- 10824270 TI - The effect of 3D structure on motion segmentation. AB - A smooth surface imaged on the retina produces a smooth flow field. Thus, the visual system may group regions of smoothly varying flow to segment surfaces. We tested this idea by having observers perform a segmentation task on several stimuli that differed in their 3D interpretations but were all matched in the smoothness of their 2D flow fields. Performance varied across conditions with the best performance occurring when the stimulus stimulated a rigid plane. This result suggests that while observers may use deviations from smoothness to segment a broad class of motion stimuli, they use a more precise strategy to segment stimuli with a familiar 3D interpretation. PMID- 10824271 TI - Transient deficit hypothesis and dyslexia: examination of whole-parts relationship, retinal sensitivity, and spatial and temporal frequencies. AB - A defect affecting the transient visual sub-system is believed to be one of the prime factors affecting reading disability. In this study, the transient deficit hypothesis was tested using the global precedence paradigm, examining retinal sensitivity, and comparing of patterns of responses to large versus small stimuli. Participants were three groups of dyslexic, chronologically age-matched, and reading age-matched children. The results revealed that although dyslexic individuals did not show any deficit in processing (a) wholes and parts (Experiment 1); (b) information in peripheral locations of the retina (Experiment 2); and (c) various sizes of the stimulus (Experiment 3); they showed a deficit in temporal processing of visual information. These findings challenge the transient deficit hypothesis in the sense that the transient sub-system has been suggested to be associated with processing of the global level, low spatial frequencies, and peripheral vision; however, they confirm that hypothesis in the sense that this visual sub-system is suggested to be associated with processing of high temporal frequencies. Transient deficit hypothesis and dyslexia: examination of whole-parts relationship, retinal sensitivity, and spatial and temporal frequencies. PMID- 10824272 TI - 1999 Seymour B. Sarason Award. Community narratives: tales of terror and joy. AB - Inspired by a literary-feminist reading of biblical texts, it is suggested that the mission of community psychology/social science can be understood as a calling to use our tools (research methods, critical analysis and observation, scholarship, social influence) to assist others in the job of turning tales of terror into tales of joy. Such work is the essence of personal and social change. These are not things we can do alone; they require collaboration between us and the people of concern. Applying concepts from narrative theory, including description and critical analysis of community and setting narratives, dominant cultural narratives, and personal stories, we can learn from our own communities and we can use our resources to help make known (perhaps even help to imagine new) tales of joy. Some of these themes are illustrated in three very different contexts: a religious community that has made itself inclusive of gay and lesbian people, a mutual help organization that offers a sense of community and hope for the future to people with a history of serious mental illness, and a public elementary school. PMID- 10824273 TI - Making a difference: the social ecology of social transformation. AB - A multidisciplinary and multilevel framework for social transformation is proposed, encompassing four foundational goals: capacity-building, group empowerment, relational community-building, and culture-challenge. Intervention approaches related to each goal are presented at the setting, geographic community, and societal levels. Four exemplars of social transformation work are then discussed: the Accelerated Schools Project, Meyerhoff Program, ManKind Project, and women's movement. These examples illustrate the synergistic relationship among the four transformational goals, within and across levels of analysis, which is at the heart of the social transformation process. The paper concludes with three challenges to guide our efforts as we enter the new century: (1) to move social transformation to the center of our consciousness as a field; (2) to articulate jointly with allied disciplines, organizations, and citizen groups an encompassing, multidisciplinary, and multilevel framework for social transformation; and (3) to do the above with heart, soul, and humility. PMID- 10824274 TI - Normative influences on aggression in urban elementary school classrooms. AB - We report a study aimed at understanding the effects of classroom normative influences on individual aggressive behavior, using samples of 614 and 427 urban elementary school children. Participants were assessed with measures of aggressive behavior and normative beliefs about aggression. We tested hypotheses related to the effects of personal normative beliefs, descriptive classroom norms (the central tendency of classmates' aggressive behavior), injunctive classroom normative beliefs (classmates' beliefs about the acceptability of aggression), and norm salience (student and teacher sanctions against aggression) on longitudinal changes in aggressive behavior and beliefs. injunctive norms affected individual normative beliefs and aggression, but descriptive norms had no effect on either. In classrooms where students and teachers made norms against aggression salient, aggressive behavior diminished over time. Implications for classroom behavior management and further research are discussed. PMID- 10824276 TI - The latent structure of substance use among American Indian adolescents: an example using categorical variables. AB - Researchers frequently have only categorical data to analyze and cannot, for theoretical or methodological reasons, assume that the observed variables are discrete representations of an underlying continuous variable. We present latent class analysis as an alternative method of measuring latent variables in these circumstances. Latent class analysis does not require the assumptions of factor analyses about the nature of manifest and latent variables, but does allow the use of more precise model selection than techniques such as cluster analysis. We modeled the lifetime substance use of American Indian youth. The latent class model of American Indian teenagers' substance use had four classes: Abstaining, Predominantly Alcohol, Predominantly Alcohol and Marijuana, and Plural Substance. We then demonstrated the usefulness of this latent variable by using it to differentiate levels of several variables in a manner consistent with Social Cognitive Theory. PMID- 10824275 TI - The relation of perceived neighborhood danger to childhood aggression: a test of mediating mechanisms. AB - In the current study, two mediational mechanisms, parenting practices and children's beliefs about aggression, were hypothesized to account for the relationship between perceived neighborhood danger and childhood aggression. Using structural equation modeling, data were analyzed from an inner-city school based sample of 732 predominantly African American 5th graders. Results suggested that perceived neighborhood danger was associated with strong positive beliefs about aggression, which in turn was associated high levels of aggression. The hypothesized mediating role of parenting practices (restrictive discipline, parental monitoring, and parental involvement) on the relation between perceived neighborhood danger and child aggression was not supported. However, the current findings suggest that children's positive beliefs about aggression mediated the relationship between restrictive discipline and aggression. Directions for future research are discussed. PMID- 10824277 TI - Partner violence, social support, and distress among inner-city African American women. AB - This study examined the role of social support in the partner violence psychological distress relation in a sample of African American women seeking medical care at a large, urban hospital (n = 138). Results from bivariate correlational analyses revealed that partner violence was related to lower perceived social support and greater psychological distress, and lower social support was related to more distress. Furthermore, findings based on path analysis indicated that low levels of social support helped account for battered women's increased distress. Findings point to the need for service providers to screen for partner violence in nontraditional sites, such as hospital emergency rooms, and to address the role of social support resources in preventive interventions with African American battered women. PMID- 10824278 TI - Cervicogenic headache: criteria, classification and epidemiology. AB - The concept that headache might stem from the neck is old. The term "cervicogenic headache" was coined in 1983. A new content was then given to this concept: cervicogenic headache (CEH) is in principle a unilateral headache, generally starting in the neck and "spreading" forwards. A strict unilaterality--that is, absolutely no pain on the opposite side--is rather rare. Unilaterality in this context is defined as follows: the headache dominates on one side. When weak, the pain may be only on that side; when severe, it may also be felt on the contralateral side, but to a lesser extent. It never dominates on the contralateral side. These special features of CEH cannot be emphasised strongly enough. There are signs pertaining to the neck, such as reduced range of motion in the neck, mechanical precipitation mechanisms and ipsilateral shoulder/arm sensation (or even pain). Migraine without aura symptoms are less prominent than in migraine. PMID- 10824279 TI - Cervicogenic headache: clinical aspects. AB - Cervicogenic headache (CEH) is a relatively common but often overlooked disorder. There is sufficient evidence to support this category and the existing diagnostic criteria are adequate. Subgroups may exist and the clinical picture sometimes may be similar to that of other headache disorders, however. The pathophysiology of this condition and its relationship with other headache syndromes remain to be determined. Migrainous features may occur in some patients. PMID- 10824280 TI - Unilateral headaches and their relationship with cervicogenic headache. AB - The concept of headache originating/starting in the neck is revised and considered in the light of previous descriptions of syndromes and entities and with reference to the current diagnostic systems for the classification of headache and other head pain. Cervicogenic headache (CEH), a clinical picture recently described by Sjaastad and coworkers and listed in the International Association for the Study of Pain (IASP) Classification, is analyzed, also taking into consideration its diagnostic criteria in terms of sensitivity and specificity. The problem of a differential diagnosis with migraine, tension headache and other well defined forms of unilateral headaches is discussed with reference to a case series of 114 patients who were selected based on their adherence to two fundamental criteria: (i) side-locked unilaterality of pain; and (ii) pain starting in the neck and spreading to the fronto-orbital area. Based on the results, these simple criteria can contribute to a preliminary identification of possible CEH cases that may then undergo a sequence of clinical and instrumental procedures in order to confirm the diagnosis and, possibly, to localize the level(s) of dysfunction in the cervical spine which may be the target for therapeutic investigations, whether invasive or non-invasive. PMID- 10824281 TI - Cervicogenic headache: current concepts of pathogenesis related to anatomical structure. AB - A brief summary will be given of the putative etiological factors/topography that so far have been discussed in the context of cervicogenic headache (CEH), in approximately sequential order. Some of these factors have some basis in clinical research and practice; others are more speculative. PMID- 10824282 TI - Pain and functional disability. AB - The pain and functional disability in headache is tentatively presented with respect to the displayed causal relationships, beginning with the nociceptor. The concept is presented that primary deep pain must stem from a pain generator (PG), of which the free C-fiber ending together with the extracellular space constitute the (inflammatory) nociceptor; and that neurogenic pain is a secondary, enhancing measure. Headache is considered to be a result of various PGs in pain-sensitive structures implicated in head locomotion. Headache occurs when nociceptive volleys from either the cephalic or cervical levels traverse the locomotor centres of the head in the upper cervical medulla and are realized as referred pain. Some clinical aspects are presented. PMID- 10824283 TI - Whiplash injuries: clinical picture and diagnostic work-up. AB - The term "whiplash" commonly refers to symptoms and signs associated with a mechanical event such as a sudden acceleration and deceleration of the neck (due, in the majority of cases, to a road accident), instead of to the mechanism itself. The recent Quebec Classification of Whiplash Associated Disorders (WAD) contributed to define nosographically all the clinical manifestations usually grouped under the terms acute/post-traumatic and late "syndrome". In the late phase of WAD, neck pain and neck muscle contraction have been reported in all cases, together with headache in over 50%. "Headache stemming from the neck", despite numerous attempts to classify this entity (i.e. cervicogenic headache) according to the IASP classification (headache associated with neck disorders), is still a subject of debate. An adequate multiparametric procedure is required to study WAD, which takes into account: the patient's principal details; an exact reconstruction of the event; description and analysis of the signs and symptoms, with various complications and correlated dysfunctions; an objective neurological and neck-shoulder examination; and a battery of complementary instrumental tests which are described in this study. These investigations include evaluation of muscle tension (manual palpation, algometry, EMG recording), kinematic analysis of the cervical spine, neuropsychological and psychological evaluation, and evaluation of disability. In order to assess cervical spine mobility in WAD patients, a 3D kinematic analysis by means of the ELITE system and clinical evaluation were performed in our setting. Seventy patients with whiplash injury and 46 healthy volunteers were enrolled in the study. Patients were tested at the time of first consultation and again 6 months and 12 months later. Clinical evaluation of the range of motion was performed both in patients and in 41 healthy volunteers. Furthermore, patients diagnosed according to the WAD Classification as grade 2 (n = 68) or grade 3 (16) underwent a Quality of life (QoL) evaluation, measured using the short form (36-item) Health Survey (SF36) and the migraine-specific questionnaire (MSQ). According to our data, whiplash patients showed an impairment of cervical spine mobility, as well as a poor QoL, compared to a control group population, even though we observed a trend towards improvement over time in cervical ROM. PMID- 10824284 TI - Symptomatic cervicogenic headache. AB - Cervicogenic headache is a little-known clinical condition whose true importance has only recently been recognized. A number of causes may lie at the basis of the onset of headache (symptomatic cervicogenic headache). However, despite exhaustive attempts, sometimes it is not possible to identify a clear cause responsible for the onset of the syndrome (primitive cervicogenic headache). The genesis of symptomatic cervicogenic headaches sometimes may be easy to identify as a result of a close, pre-existing, cause-effect relationship (i.e. trauma). On other occasions it may be much more laborious to pinpoint the pathology responsible for headache (some cranio-cervical anomalies, etc.). Clinically, it is necessary to perform a thorough preliminary clinical and anamnestic evaluation which can orient subsequent investigations to achieve a diagnosis in the least time possible with the minimum discomfort to the patient and his relatives, not to mention lower costs for society. PMID- 10824285 TI - Proinflammatory pathways in cervicogenic headache. AB - Cervicogenic headache (CEH) is a relatively common form of headache arising from the neck structures. The pathophysiology probably results from various local pain producing factors such as intervertebral dysfunction, with a no less important role played by the frequent coexistence of a history of head traumas. This report represents a series of pathophysiological studies in CEH patients and the results achieved by pharmacological treatment of the disease. Interleukin-1 beta (IL-1 beta) and Tumour Necrosis Factor alpha (TNF-alpha) exert their multifunctional biological effects by promoting and increasing the molecular events of cellular inflammation. We found that the cytokine pattern of CEH patients is--similar to cluster headache--biased towards an inflammatory status. Higher levels of both IL 1 beta and TNF-alpha were detected in the sera of CEH patients than the levels in patients with migraine without aura and in healthy subjects. There were also differences between the spontaneous and mechanically worsened pain phases of CEH. Nitric oxide (NO) synthase is also activated in cervicogenic headache. No change in NO metabolites levels has been observed after NO donor administration. This behaviour is clearly different from that observed in migraine and tension headache patients. We conclude that the high degree of cytokine and NO production in CH may depend on the differing pathophysiological mechanisms at work in CEH than in other forms of headache. PMID- 10824286 TI - Cervicogenic headache: techniques of diagnostic nerve blocks. AB - Ther term cervicogenic headache (CEH) was introduced by Sjaastad and co-workers in 1983. In 1990 Sjaastad et al. published diagnostic criteria for CEH. In 1998 refinements of these criteria were published, emphasising the use of diagnostic nerve blocks in patients with CEH as important confirmatory evidence. However, the standardisation of diagnostic nerve blocks in the diagnosis of CEH remains to be defined. Herein we present an overview of diagnostic nerve blocks in the cervical area. Suggestions as to their role in the diagnosis of CEH are given. PMID- 10824287 TI - Current methods for cervical spine movement evaluation: a review. AB - Cervical spine mobility is difficult to investigate accurately because of its anatomic structure and the compensatory movements. Different methods have been conceived in order to obtain a reliable measurement of cervical range of movement (ROM). We reviewed different instruments described in the literature: x-rays, CT and MRI, goniometer, inclinometer, cybex and related devices, and opto-electronic scanners. Cybex and 3D kinematic analysis by means of opto-electronic scanners (Elite system) seemed to be the most reliable and reproducible methods. Cybex equipment is relatively inexpensive and easy to use in a clinical setting, while the Elite system is expensive and requires special training of the personnel. However, the choice of method depends primarily on whether the physician's goal is a clinical screening or a thorough investigation of neck function (e.g., post traumatic cervical spine disorders). For the first purpose, certain types of goniometers (gravity goniometer, ad modum Myrin), as well as the cybex, show good reproducibility and reliability in evaluating maximal cervical ROM (flexion extension, rotation, lateral bending), while x-rays and, above all, 3D kinematic analysis (using opto-electronic scanners) are more suitable for diagnostic and follow-up evaluation of neck disorders. PMID- 10824288 TI - Cervicogenic headaches: radiofrequency neurotomy and the cervical disc and fusion. AB - OBJECTIVE: Headaches that originate from pathology of the cervical spine, called "cervicogenic headache", have been debated, described, and treated by various researchers. This paper describes the use of radiofrequency neurotomy procedures to relieve cervicogenic headache at several strategic locations. Procedures listed include those to the greater occipital nerve territory, the C2 medial rami, and the cervical discs. Anatomy relevant to the innervation of the disc by way of the sympathetic chain is described and illustrated. METHODS: Radiofrequency neurotomy procedures were performed following either a successful nerve block in either the distribution of the greater occipital nerve or the C2 medial rami, or after provocative cervical discography. Cervical disc and fusion surgery is being correlated to the nerve supply of the discs and ligaments. RESULTS: The majority of patients suffering from cervicogenic headaches can be totally relieved for a lifetime, especially if the pain is unilateral. A very, very small percentage of these patients cannot be helped. The remaining sufferers have a considerable reduction in the intensity and frequency of pain. DISCUSSION: Each of the procedures discussed, or a combination of all of them, can alleviate cervicogenic headaches completely, as is our goal. PMID- 10824289 TI - Cervicogenic headache: prevalence and response to local steroid therapy. AB - Cervicogenic headache (CEH) has been said to be common among patients with idiopathic headache, but no information exists as to its prevalence among those who have not suffered whiplash or head injury. This study was designed to answer this question and in addition to determine whether headache relief could be achieved by blockade of the occipital nerves (greater and lesser occipital--GON, LON) in the upper neck, on the side habitually affected by the headache. Among 796 patients with idiopathic headache, 128 or 16.1% were found to be suffering from CEH. They were predominantly female, as in the case of migraine, older than the migrainous group (49.5 years as against 34.7 years), respectively, and with a monthly headache frequency of 18, against 6.9 in the case of migraine. Injections of depot methylprednisolone into the region of the GON and LON produced complete relief of headache in 169 out of 180 patients with CEH for a period ranging from 10 to 77 days, the mean duration of relief being 23.5 days. However, similar relief of headache could be achieved in patients with attacks of strictly unilateral migraine or cluster headache, suggesting that local steroid injections by blocking the cervico-trigeminal relay, can arrest other forms of unilateral headache. PMID- 10824290 TI - Epidural steroids as a pharmacological approach. AB - Cervicogenic headache is a relatively common pain syndrome related to functional and/or degenerative alterations of the cervical spine tract. Administration of steroid represents an effective therapy for this headache, due to the anti inflammatory effects combined with its direct analgesic effects on the C fibers. The epidural injection of steroids, while requesting skilled personnel for its execution, gives short term (2-month) pain relief with few risks or side effects. Moreover, epidural steroids allow reduction of analgesic drug consumption. PMID- 10824291 TI - Surgical treatment of non-responsive cervicogenic headache. AB - OBJECTIVE: 102 patients suffering from long-lasting, very severe cervicogenic headache (CEH), non-responsive to physical or drug therapy, were surgically treated. METHODS: Different diagnostic procedures and their significance for determining the advisability of surgery are summarised. The particular importance of the patient history and local anaesthesia together with the clinical examination is outlined. Different surgical approaches are described: ganglionectomy, ventral and dorsal decompressive operation. RESULTS: CEH can be triggered by vascular or scar tissue compression of the C2 root and ganglion and irritation of other upper cervical nerve roots (C3, C4). Vascular compression is caused by: (a) the sinusoidal venous plexus, which surrounds the ganglion and nerve root like a cuff and may be dilated upon raised venous pressure; (b) further on by arterial loops throbbing against the ganglion; and (c) (rarely) by arteriovenous (AV) malformations. Nerve fibre degeneration is demonstrated morphologically by electron optical investigation. Afferences from ganglion C2 to the brain stem, as documented by experimental investigation on cats using the injection of HRP into the C2-ganglion, can explain the reference of pain from the neck to the fronto-ocular region and could at the same time elucidate the genesis of accompanying symptoms. Degenerative diseases such as disc protrusion and retrospondylosis have been shown to be other trigger mechanisms evoking CEH, as is well known from facet joint arthrosis. Degenerative diseases usually cause dura compression with narrowing of the spinal canal and frequently, in addition, instability. Evocation of CEH could be explained by the irritation, by those degenerative diseases, of structures with pain-conducting nerve fibres (facet joint capsule, nerve root, longitudinal ligaments, spinal dura, disc). About 80% of our surgically treated patients were relieved of pain or improved during a long period of follow up. The recurrence of degenerative alterations with new irritation from pain-conducting structures is thought to be responsible for the recurrence of headache. Further surgical approaches for the treatment of patients with the recurrence of pain are discussed. CONCLUSION: Various surgical treatments are suggested to treat long-lasting severe CEH in patients not responsive to any physical or drug therapy. PMID- 10824292 TI - Novel anticonvulsants: a new generation of mood stabilizers? AB - Accumulating evidence suggests that at least some novel anticonvulsants may have mood-stabilizing properties. This paper reviews the literature for empirical studies of this topic. Lamotrigine has the most evidence in favor of its efficacy, with two double-blind studies in which it was more efficacious than placebo in the treatment of bipolar depression. However, it is associated with a 1/1000 risk of potentially fatal Stevens-Johnson syndrome. Gabapentin, although safe and well-tolerated, has been found in two double-blind studies not to be efficacious in treatment-refractory mania or refractory bipolar depression. Topiramate is currently supported only by naturalistic evidence of mild to moderate mood-stabilizing efficacy, but it has the advantage of often producing weight loss. Based on these data, lamotrigine may be effective, in monotherapy or as an adjunct, for treating depression in type I bipolar disorder, but suggestions regarding gabapentin and topiramate await further efficacy data. Most of the current findings derive from small, non-double-blind studies, and further research is required before clinicians can consider any of these agents to be mood stabilizers. PMID- 10824293 TI - Dysphoric disorders and paroxysmal affects: recognition and treatment of epilepsy related psychiatric disorders. AB - Interictal dysphoric disorder is an intermittent and pleiomorphic affective somatoform disorder that presumably occurs as a result of inhibitory mechanisms in chronic mesial temporal lobe epilepsy. Treatment with antidepressant medication, enhanced if necessary with small doses of an atypical antipsychotic, tends to be highly effective. The dysphoric disorder also occurs in the absence of epilepsy in a subictal variation, particularly in patients with brain lesions and as premenstrual dysphoric disorder. The paroxysmal affects, ranging from irritability through anger to rage, play a major role in interictal dysphoric disorder. Their manifestation among patients with mesial temporal lobe epilepsy is counterbalanced by the fact that these individuals tend to be highly ethical and religious. The paroxysmal affects that may emerge with vehemence during episodes of interictal dysphoric disorder play a role in all people, differing in prominence among individuals. For a comprehensive view of the psychiatric aspects of epilepsy, the important premodern findings are reviewed together with recent ones. PMID- 10824294 TI - Theoretical and methodological issues in psychiatric comorbidity. AB - The occurrence of multiple diagnoses in one patient is a phenomenon of major clinical and theoretical importance. This paper reviews the various factors involved in real and artifactual comorbidity. Important causes of spurious comorbidity are discussed, including invalidity of the individual diagnoses, use of inappropriate diagnostic paradigms, descriptive overlap of diagnostic criteria, ascertainment bias, and diagnostic bias. To illustrate some of the concepts discussed, two examples are presented: the comorbidity of schizophrenia and substance use disorders, and the comorbidity of posttraumatic stress disorder and major depression. The study of comorbidity can advance psychiatry by helping us to clarify our thinking about categories of illness and the boundaries between them, as well as the relationships among these categories. PMID- 10824295 TI - When goodbye precedes hello: a "premature" termination. PMID- 10824296 TI - Impaired physicians: is there a duty to report to state licensing boards? PMID- 10824297 TI - Psychotic depression: what is it and how should we treat it? PMID- 10824298 TI - Cardiovascular fitness in firefighters. PMID- 10824299 TI - Race, gender, and social status as modifiers of the effects of PM10 on mortality. AB - Interest has recently been focused on which populations are most at risk of premature mortality induced by air pollution. This coincides with greater concern about environmental justice. We analyzed total mortality in the four largest US cities with daily measurements of particulate matter less than 10 microns (PM10) and combined the results to determine whether race, sex, and education are potential modifiers of the effects of PM10 on mortality. We computed daily counts of deaths stratified by sex, race, and education in each city and investigated their associations with PM10 in a Poisson regression model. We combined the results by using inverse variance weighted averages. We found evidence of effect modification by sex, with the slope in female deaths one third larger than in male deaths, whereas for social factors and race we found only weak evidence of effect modification. In general, the effect modification appeared modest compared with other reports of substantial effect modification by medical conditions. PMID- 10824300 TI - Measurement of percutaneous uptake of 2-methoxy ethanol vapor in humans. AB - Absorption of vapors through skin has been largely ignored in occupational health, although for ethylene glycol ethers this route of exposure could be more important than inhalation. We used an automated concentration and humidity controlled system to measure real-time percutaneous absorption of 2-methoxy ethanol (ME) vapor in seven volunteers. The exposure concentration (300 +/- 10 ppm or 25 +/- 0.5 ppm), humidity (80 +/- 2%), and temperature (27.5 +/- 0.5 degrees C) were controlled throughout the experiments. Uptakes during 4-hour single-arm exposure at 25 ppm and 300 ppm were 7.0 mg and 65.3 +/- 25.0 mg, respectively, with corresponding uptake rates of 1.36 micrograms/cm2/hr and 13.2 +/- 5.0 micrograms/cm2/hr. Percutaneous absorption was consistent and unsaturated during exposure. Because the permeability constant of ME vapor (14.0 +/- 5.3 cm/hr) was much higher than that of many widely used organic chemicals, we concluded that vapor absorption through skin is a significant contributor to overall ME exposure. PMID- 10824301 TI - Intelligent database generated occupational questionnaire system. AB - Obtaining an adequate occupational history requires special expertise to "ask the right questions" that are relevant to a particular patient's specific health conditions and potential exposures. This article describes a way to systematically accomplish this by means of a computer system that can overcome limited availability of necessary clinical occupational health expertise. The Intelligent Questionnaire system is a computer-based system for generating case specific questionnaires about the influence of work on respiratory disease. Intelligent Questionnaire includes three databases: Questions, Responses, and Calls (clues to identify questions). The Questionnaire also arranges questions in a logical manner and provides a customized data entry screen for each subject. This approach provides primary practitioners with expertise on a case-by-case basis. It also facilitates occupational health surveillance because it allows acquiring detailed case-specific information in a systematic fashion. A computer based system can facilitate obtaining occupational histories with high specificity and consistency without depending on general availability of a human occupational health clinical expertise. PMID- 10824302 TI - Illnesses among United States veterans of the Gulf War: a population-based survey of 30,000 veterans. AB - Despite numerous studies on veterans of the 1990 to 1991 Gulf War, the fundamental questions of how healthy they are and how their health compares with that of their military peers who were not deployed to the Gulf have not been fully answered. We conducted a health survey in which the health outcomes of a population-based sample of 15,000 Gulf veterans representing various military branches and unit components (regular, reserve, National Guard) were compared with those of 15,000 non-Gulf veterans who were randomly sampled to mirror the number in the same military strata in the Gulf veteran group. In comparison with their peers, Gulf veterans had a higher prevalence of functional impairment, health care utilization, symptoms, and medical conditions and a higher rate of low general health perceptions. A longitudinal follow-up of the health of these veterans will be needed to detect changes in health status and to detect diseases with a long latency period. PMID- 10824303 TI - The impact of the Citibank, NA, health management program on changes in employee health risks over time. AB - This study estimated the impact of the Citibank Health Management Program on changes in health risks among Citibank employees. McNemar chi-squared tests compared the probability of being at high risk for poor health when the first and last health-risk appraisal surveys were taken. Logistic regression controlled for baseline differences in subsequent analyses when those who participated in more intensive program features were compared with those who participated in less intensive features. Declines in risk were noted for 8 of 10 risk categories. Most changes were small, except those related to exercise habits, seatbelt use, and stress levels. For nine health risk categories, those who participated in more intensive program services were significantly more likely than others to reduce their health risks. Thus, the Citibank Health Management Program was associated with significant reductions in health risk. PMID- 10824304 TI - Provocation with stress and electricity of patients with "sensitivity to electricity". AB - Twenty-four patients with self-reported "sensitivity to electricity" were divided into two groups and tested in a double-blind provocation study. These patients, who reported increased skin symptoms when exposed to electromagnetic fields, were compared with 12 age- and sex-matched controls. Both groups were exposed to 30 minute periods of high or low stress situations, with and without simultaneous exposure to electromagnetic fields from a visual display unit. The matched controls were tested twice and given the same exposure as the patients but had the fields turned on every time. Stress was induced by requiring the participants to act in accordance with a random sequence of flashing lights while simultaneously solving complicated mathematical problems. Blood samples were analyzed for levels of the stress-related hormones melatonin, prolactin, adrenocorticotrophic hormone, neuropeptide Y, and growth hormone, and the expression of different peptides, cellular markers, and cytokines (somatostatin, CD1, factor XIIIa, and tumor necrosis factor-alpha). Skin biopsies were also analyzed for the occurrence of mast cells. Stress provocation resulted in feelings of more intense mental stress and elevated heart rate. The patients reported increased skin symptoms when they knew or believed that the electromagnetic field was turned on. With the blind conditions there were no differences between "on" or "off." Inflammatory mediators and mast cells in the skin were not affected by the stress exposure or by exposure to electromagnetic fields. The main conclusion was that the patients did not react to the fields. PMID- 10824305 TI - Occupation and prostate cancer risk in Sweden. AB - To provide new leads regarding occupational prostate cancer risk factors, we linked 36,269 prostate cancer cases reported to the Swedish National Cancer Registry during 1961 to 1979 with employment information from the 1960 National Census. Standardized incidence ratios for prostate cancer, within major (1 digit), general (2-digit), and specific (3-digit) industries and occupations, were calculated. Significant excess risks were seen for agriculture-related industries, soap and perfume manufacture, and leather processing industries. Significantly elevated standardized incidence ratios were also seen for the following occupations: farmers, leather workers, and white-collar occupations. Our results suggest that farmers; certain occupations and industries with exposures to cadmium, herbicides, and fertilizers; and men with low occupational physical activity levels have elevated prostate cancer risks. Further research is needed to confirm these findings and identify specific exposures related to excess risk in these occupations and industries. PMID- 10824306 TI - Flexor tendon entrapment of the digits (trigger finger and trigger thumb). AB - Flexor tendon entrapment of the digits is a disorder characterized by snapping or locking of the thumb or fingers (with or without pain). Most cases are secondary to thickening of the digit's A1 pulley, but other pathogeneses include tendon abnormalities at the level of the carpal tunnel, thickening of other pulleys, and abnormalities of the metacarpal-phalangeal joint. Its historical name, stenosing tenosynovitis of the digits, is inappropriate because histological studies document a lack of inflammation. Flexor tendon entrapment of the digits is a relatively common, uncomplicated, and non-controversial musculotendinous disorder of the distal upper extremity. The purpose of this invited review is to summarize information from the medical literature on aspects of this condition likely to be of interest and relevant to occupational medicine practitioners. Topics covered include normal anatomy and kinesiology, history, clinical observations related to diagnosis, pathology, pathophysiology, clinical observations on etiology, descriptive epidemiology, epidemiological studies, and case management. Models for the pathogenesis of flexor tendon entrapment of the digits are proposed, and opportunities for future research are presented. PMID- 10824307 TI - Hazardous materials events: an industrial comparison. AB - Identifying industries at high risk for hazardous materials releases can facilitate prevention and preparation for such events. A retrospective review by Standard Industrial Classification (SIC) codes was conducted on non-petroleum hazardous materials emergency events from 1993 to 1995 and collected by the Washington State Department of Health in a program supported by the Agency for Toxic Substances and Disease Registry. Annual US Census data were used to determine the number of facilities and employees by SIC code in the state of Washington. SIC codes with the most total events and events per 10,000 employees were ranked and characterized by type of releases. In 3 years, 1269 events were recorded, with 294 involving human victims. Industries with the highest average annual number of events per 10,000 employees were agricultural chemical manufacturing (142); petroleum refining (122); industrial and miscellaneous chemical manufacturing (56); electric light and power (54); and pulp, paper, and paperboard mills (39). Industries with high rates of hazardous materials emergency events should continue to develop methods of preventing these releases. PMID- 10824308 TI - Non-Hodgkin's lymphoma and exposure to benzene in a multinational cohort of more than 308,000 petroleum workers, 1937 to 1996. AB - Petroleum workers are exposed to benzene or benzene-containing petroleum products. As such, studies of these workers provide an opportunity for investigating the relationship between benzene and non-Hodgkin's lymphoma (NHL). However, few cohort studies of petroleum workers report results of NHL separately. One reason is that NHL is usually grouped with other lymphopoietic cancers in the analysis. Another reason is the relatively small number of NHL cases in some studies. To determine the risk of NHL in petroleum workers, we identified 26 cohorts of petroleum workers in the United States, the United Kingdom, Canada, Australia, Italy, and Finland. Authors of the original studies were contacted, and data on the number of observed deaths and person-years of observation were requested. Data from these studies were reviewed individually as well as combined in a pooled analysis (meta-analysis). In particular, results for individual cohorts, most of which had never been reported before, were presented. The combined multinational cohort consisted of more than 308,000 petroleum workers (6.6 million person-years), and the observation period covered an interval of 60 years from 1937 to 1996. A total of 506 NHL deaths were observed, compared with 561.68 expected. The standardized mortality ratio was 0.90 and the 95% confidence interval was 0.82 to 0.98. Analyses were performed by type of facility and industrial process. Stratum-specific standardized mortality ratios (95% confidence intervals) were 0.96 (0.86 to 1.07) for US refinery workers, 1.12 (0.90 to 1.37) for non-US refinery workers, 0.64 (0.50 to 0.82) for product (gasoline) distribution workers, and 0.68 (0.47 to 0.95) for crude oil workers. When individual cohorts were stratified by length of observation, no pattern was detected. In general, exposure levels before 1950 were much higher than thereafter. However, analysis of workers by hire date (< 1950, > or = 1950) revealed no difference in NHL mortality. Furthermore, none of the individual studies showed significant exposure-response relations. In summary, results from individual studies, as well as from the pooled analysis, indicated that petroleum workers were not at an increased risk of NHL as a result of their exposure to benzene or other benzene-containing petroleum products in their work environment. This conclusion was supported by cohort studies of workers in other industries who were exposed to benzene as well as by population-based case-control studies of NHL and occupational exposures. PMID- 10824309 TI - Telling stories: the health benefits of narrative. PMID- 10824310 TI - Criminal language and poetic jailbreak: writing chronic fatigue immune dysfunction syndrome. PMID- 10824311 TI - Story language: a sacred healing space. PMID- 10824312 TI - Pretty reflections. PMID- 10824313 TI - The hospital prayer book: a partner for healing. PMID- 10824314 TI - Circles. PMID- 10824315 TI - Approaching death by writing: Montaigne's Essays and the literature of consolation. PMID- 10824316 TI - Truth, trauma, and justice in Gillian Slovo's Every Secret Thing. PMID- 10824317 TI - Regression and psychoanalytic technique: the concretization of a concept. AB - An exploration of the regression concept historically and conceptually reveals that its familiarity and frequent use have resulted in decreasing conceptual clarity and precision. Rooted in an outmoded fixation-regression model of development and psychopathology, the concept has become concretized. This paper is a beginning exploration of problematic aspects of the concept of regression, with emphasis on potentially detrimental consequences for psychoanalytic technique that derive from its unexamined use. Some of the salient issues are illustrated with clinical examples. PMID- 10824318 TI - Modern hermeneutics and psychoanalysis. AB - Contemporary hermeneutics tries to integrate our unique, local sense of things with overarching nature, often by celebrating the concrete phenomenology of the moment at the expense of scientific abstractions. But abstractions are unavoidable. Hermeneuticists point out that we are constantly making new abstractions. But the more optional and variable views, which we call subjective, depend on the old, reliable abstractions, such as time, space, substance, and causality, that constitute our fixed reality. Hermeneutics usefully challenges psychoanalysis to justify its way of slicing up the mind and treatment process. PMID- 10824319 TI - From schisms through synthesis to informed oscillation: an attempt at integrating some diverse aspects of psychoanalytic technique. AB - There exists a conceptual bifurcation at the core of psycho-analysis. It has been viewed from differing vantage points and portrayed as subsuming various dichotomies (oedipal-preoedipal, conflict-deficit, one person-two person, classic romantic, and so on). While each such conceptual pair has its own heuristic accompaniments, these dichotomies share a profoundly important element. They have divergent effects upon the analyst's mode of listening and the nature of his or her interventions. These and other related technical implications are the topic of this paper. With the help of three clinical vignettes and by coalescing the isolated voices of many distinguished theoreticians, the author attempts to elucidate and heal this split. This paper proposes three levels of increasingly sophisticated resolution of the technical divergence resulting from this schism. The paper recommends an informed oscillation between the two polarities of psychoanalytic technique, an oscillation that must remain in consonance with the patient's shifting ego organization. The paper concludes by highlighting the developmental bases for the proposed technical conceptualizations. PMID- 10824320 TI - Representations of dread: the dreaded self and the dreaded state of the self. AB - The experience of dread, an extreme form of fear that is induced by terror and horror, is seen as manifested in the shapes of a "dreaded self" and a "dreaded state of the self." These representations reflect psychic dangers ranging from a common, feared identification to states of disconnection, desolation, ego dissolution, and nonexistence. It is suggested that life crises and traumatic impingements, informed by developmental and psychic realities, are critical determinants of multiple dreaded self-representations; that disavowal often serves to massively ward off the recognition of the awful; and that these representations serve a preconscious signal function that anticipates the danger of reexperiencing an original terror. Case examples illustrate these points and reflect the utility of the language of dreaded representations in the treatment situation. PMID- 10824322 TI - A structural and intertextual reading of Freud's "On Dreams". AB - By using the framework of a "quest" narrative based on literary allusions to Virgil's Aeneid and Goethe's Wilhelm Meister's Apprenticeship, Freud transformed the 500 pages of The Interpretation of Dreams into some fifty pages entitled "On Dreams." This paper elucidates the narrative means by which Freud achieved the feat of turning a highly complex, lengthy theoretical work into an engaging narrative. Its main plot, provided by a specimen dream, reveals Freud's working through of the personal and professional conflicts of his life up to 1901, and serves as a practical demonstration of and paradigm for the process of working through in psychoanalysis. PMID- 10824321 TI - On analytic listening. AB - Multiple dimensions of the listening process as implemented in the analytic process are discussed. Listening is not the same as hearing; it is done with the mind rather than just the ears. Listening seeks meaning, specifically the meaning in the mind of the patient. The meaning of words is often obscure, ambiguous, and uncertain, and their deeper implications can only be approached over time through uncovering associative linkages. Listening takes place in multiple perspectives- subjective/objective, active/passive, dynamic/genetic, etc. Listening is also contextually related to dimensions of the analytic relation, including transference, alliance, and real relation. Modalities of listening related to each are explored for both analyst and analysand, and aspects of listening empathically and listening to silence are discussed. PMID- 10824323 TI - Evolution of microbiological safety cabinets. AB - Microbiological safety cabinets have had a chequered history, often being confused with chemical fume cupboards and cabinets intended to protect cultures from contamination rather than workers from infection. They have suffered from incorrect and inapt names. Much work has been required to determine the levels of air flow necessary to contain infectious particles and to prevent the exhaust of those particles to the environment, and this is considered in historical sequence. Warnings are given about the shortcomings of portable and home-made cabinets. This review traces the evolution of these cabinets, from the time of Robert Koch to the development of good national standards. PMID- 10824324 TI - Methicillin-resistant Staphylococcus aureus: evaluation of detection techniques on laboratory-passaged organisms. AB - Strains of methicillin-resistant Staphylococcus aureus (MRSA) are a major cause of nosocomial infection. This has led to the widespread introduction of screening techniques to identify patients or staff colonised with the organism and prevent the spread of MRSA in the hospital environment. The aim of laboratory techniques for the detection of MRSA screening is the rapid isolation and identification of MRSA, enabling timely implementation of appropriate infection control measures. This study compares commercially available products used by the majority of laboratories during routine screening of samples for MRSA. Several selective media are tested, including mannitol salt agar, Mueller Hinton agar, milk agar and blood agars. The accuracy of combining rapid agglutination from selective media for identification of MRSA is also determined, using six commercially produced kits. Owing to debate concerning the use of enrichment broth, in addition to primary isolation on solid media, four enrichment broths are tested and compared to direct culture techniques. Using a selection of laboratory passaged MRSA phage types, results demonstrated that mannitol salt agar containing 75 g/L NaCl, without the addition of oxacillin, recovered all 50 MRSA strains and produced the highest isolation rate. Robertson's cooked-meat broth, with 7.5% NaCl, proved the most effective enrichment medium and was more sensitive than direct culture by 3%. Pastorex Staph-plus proved to be the most efficient rapid agglutination kit when testing MRSA colonies removed directly from selective agars. PMID- 10824325 TI - Microbiological quality of prepared foods from selected kitchen outlets in Lahore, Pakistan. AB - Following UK PHLS guidelines, a total of 166 food samples from a variety of different kitchen outlets were examined for microbiological quality. Over 60% of the samples had satisfactory aerobic plate counts, and only 10% (8.5-18.7% of the samples, depending on the nature of the product) were classified as unsatisfactory. Almost 85% of samples presented with low coliform counts, and 6.6% were found to have unsatisfactory or unacceptable levels. Salmonella was found in only one sample, which had been purchased from a street trader. PMID- 10824326 TI - Oxoid CO2 Gen atmosphere generation system for growth of capnophilic bacteria: an evaluation. AB - The Oxoid CO2 Gen system is compared with BBL GasPak and a carbon dioxide (CO2) incubator to evaluate its ability to support and enhance the growth of capnophilic bacteria. Clinical samples (n = 109) from various anatomical sites and 23 spiked samples are evaluated. The criteria used to compare the systems include amount of growth, colony size, colony morphology and haemolysis. Isolation rates, amounts of growth and morphology were similar in each system, but colony diameter was significantly larger in the jar-based systems. Significantly larger colonies grew in CO2 Gen than in BBL GasPak. alpha Haemolytic zones were significantly larger in jar-based systems than in the CO2 incubator, and significantly larger in CO2 Gen than in BBL GasPak. beta haemolytic zones were significantly larger in CO2 Gen than in the CO2 incubator. The CO2 Gen system appears to be an excellent alternative to established methods for generating an environment for capnophilic incubation. PMID- 10824327 TI - Heavily pigmented melanocytic neoplasms: comparison of two melanin-bleaching techniques and subsequent immunohistochemical staining. AB - The effect of melanin bleaching using permanganate/oxalate and dilute hydrogen peroxide on subsequent immunohistochemical staining of heavily pigmented melanocytic neoplasms is investigated. Permanganate/oxalate precluded the use of some antibodies but allowed much faster bleaching times, whereas dilute hydrogen peroxide enabled a full range of antibodies to be used, yet bleaching times were far longer. Each technique has advantages; however, the choice of method should be determined by the nature of the information needed to make a diagnosis and the speed at which a report is required. PMID- 10824328 TI - Effect of haemodialysis on peripheral lymphocyte carnitine levels in patients with chronic pyelonephritis. AB - The effect of haemodialysis on peripheral blood lymphocyte levels of total carnitine (TC), free carnitine (FC) and acylcarnitine (AC), and on the ratio of AC to FC, is investigated in 20 healthy subjects (13 men and seven women) and 27 patients (10 men and 17 women) with chronic pyelonephritis. The mean predialysis peripheral lymphocyte levels of TC, FC and AC were not significantly different from those in the healthy controls (P > 0.05). However, after haemodialysis, peripheral lymphocyte levels of TC, FC and AC decreased significantly, compared with either predialysis (TC: 40%, FC: 37%, AC: 52%) or healthy controls (TC: 61%, FC: 63%, AC: 50%) (P < 0.01). Furthermore, after haemodialysis the AC/FC ratio was significantly higher, compared with either predialysis (41%) or healthy controls (38%) (P < 0.01). PMID- 10824329 TI - Monitoring human chorionic gonadotrophin level: evaluation of urine as an alternative specimen type. AB - Although urine has been used widely for the qualitative detection of human chorionic gonadotrophin (hCG), serum is chosen conventionally for the serial quantification of the hormone to monitor trophoblastic activity. In response to requests from both clinicians and patients regarding the use of urine as an alternative specimen type, we designed this comparative study to evaluate the possibility, taking into account both laboratory technique and the distribution of hCG within different body fluids. Using the Access Chemiluminescent Immunoassay System, total beta-hCG was measured in serum and urine (n = 30) collected from patients hospitalised for first-trimester abnormal pregnancy. Results obtained with normalised urine (corrected with urinary creatinine) and serum total beta-hCG correlated well (r = 0.98, P < 0.001), and we concluded that urine could be used as an alternative specimen type for the serial quantitation of hCG to monitor trophoblastic activity. However, the assay used must detect the common beta 2 epitope. PMID- 10824330 TI - Laboratory assessment of five enzyme immunoassay Clostridium difficile toxin detection kits. PMID- 10824331 TI - Neutrophil separation technique for neonates. PMID- 10824332 TI - Carcinoembryonic antigen: cell adhesion molecule and useful diagnostic marker. AB - Carcinoembryonic antigen (CEA) was first identified over 30 years ago as a marker of malignant colonic cells, but has since been shown to be expressed by a range of normal epithelial cells. Moreover, CEA is one member of a larger gene family and shares structural homology with other family molecules. Despite this confused picture, CEA is now known to function in several biological roles, including cell cell adhesion. Its use in the diagnostic setting has also changed over the years, where in routine immunohistochemistry, monoclonal antibodies to CEA are useful in aiding diagnosis when used as part of a panel. Here, recent advances in understanding the biological role(s) that CEA and CEA-related antigens may play in cell adhesion are highlighted, together with results from molecular studies that illustrate how structure can influence the choice of CEA antibodies for use in both the research and diagnostic laboratory. PMID- 10824333 TI - 11 beta-Hydroxysteroid dehydrogenase: a link between the dysregulation of cortisol metabolism and hypertension. AB - Endocrine pathology is a well-recognised and important cause of human hypertension. Recent research has highlighted the role of 11 beta-hydroxysteroid dehydrogenase (11 beta-HSD) in the development of certain forms of hypertension. This enzyme, which exists as two genetically unique isoforms, 11 beta-HSD1 and 11 beta-HSD2, is responsible for the interconversion of biologically active cortisol with its inactive 11-oxo derivative, cortisone. Congenital deficiency of 11 beta HDS2 results in inappropriate activation of the renal mineralocorticoid receptor by cortisol, leading to hypertension, hypokalaemia and metabolic alkalosis. Several authors have postulated a link between changes in 11 beta-HSD activity and the development of certain forms of essential hypertension. The existence of endogenous inhibitors of the enzyme provides compelling evidence in favour of this hypothesis, but few have been able to demonstrate a clear link between inhibition of 11 beta-HSD2 activity and hypertension by this mechanism. Similarly, several authors have suggested a relationship between reduced placental 11 beta-HSD2 activity, low birth weight with high placental weight, and the development of hypertension in adulthood. However, no clear evidence to suggest a direct correlation between birth weight, placental weight and 11 beta HSD2 activity has been demonstrated. While the role of 11 beta-HSD in the development of hypertension remains controversial, an understanding of the interplay of this enzyme with both mineralocorticoid and glucocorticoid receptors undoubtedly will yield data that will clarify this complex field. PMID- 10824334 TI - Current perspectives in evidence-based laboratory medicine. AB - This review examines some of the current perspectives in evidence-based laboratory medicine. From evidence-based medicine, which has concentrated mainly on treatments, we have learned that systematic reviews of the literature are the major way of producing high-quality evidence. This is because systematic reviews search all the literature, apply quality standards to ensure that only the best evidence is available, and aggregate a sufficient weight of information to inform on both the direction of a result (that a treatment works) and the magnitude of the result (how good the treatment is). In laboratory medicine, we have few systematic reviews. Even those we do have give little insight into features of studies which provide the closest view of the 'truth'. It may well be that current studies of laboratory tests are just not good enough. Often, they are too limited in scope, too small, and choose inappropriate patient populations. In the future, comprehensive, prospective and large studies should examine patient demographics and clinical history, as well as laboratory findings. This will better predict those features that are most closely associated with the correct diagnosis in a particular clinical situation. Clinical decision rules combining all available data will be the best way forward. PMID- 10824335 TI - Colonic perforation due to Campylobacter colitis following tonsillectomy. PMID- 10824336 TI - A time for change and a time for vision. PMID- 10824337 TI - The early detection of colorectal cancer in primary care. PMID- 10824338 TI - 'Practice professional development plans': general practitioners' perspectives on proposed changes in general practice education. AB - BACKGROUND: The Chief Medical Officer has presented a report proposing a change in general practitioners' education towards a 'Practice Professional Development Plan', which, in principle, is based around formal needs assessment, practice based learning in areas identified by those involved, and with the potential for multi-professional learning. This aims to replace the present system of a financial allowance earned by attending a certain amount of educational activity. AIM: To study the opinions of a group of general practitioners attending a course that included workshops that introduced and considered this educational initiative. METHOD: Semi-structured interviews four weeks after the course. RESULTS: Educational benefits were clearly seen, while issues such as funding and time will present difficulties in implementation. CONCLUSIONS: This proposal was seen as an improvement to the existing postgraduate educational allowance system. To maintain enthusiasm, successful introduction will depend on the issues of support and resources. PMID- 10824339 TI - Sex differences in cardiovascular disease: are women with low socioeconomic status at high risk? AB - BACKGROUND: Cardiovascular disease is still portrayed as a typical male disease, and men are more often submitted to invasive procedures or referred earlier. AIM: To explore sex differences in morbidity and referral patterns in cardiovascular disease in general practice, and the role of age and socioeconomic status. METHOD: Data were obtained from a continuous morbidity registration project in the Netherlands from 1986 to 1995 in which 12,000 patients were followed over 10 years. The effects of sex, age, and socioeconomic status on morbidity of cardiovascular disease and the referral patterns were established. RESULTS: The sex difference in morbidity becomes smaller with increasing age. Morbidity was highest in the lower socioeconomic status in general and for angina pectoris in particular. Women with angina pectoris with low socioeconomic status have a relative risk of 2.24 (CI = 1.17-3.26) compared with women with high socioeconomic status. In men, no significant difference was found between the socioeconomic status groups. For angina pectoris the sex difference in referral to the specialist was most significant: 50.6% and 26.6% (P = 0.002) for men and women respectively. CONCLUSION: For women, low socioeconomic status was associated with relatively higher morbidity of angina pectoris and myocardial infarction than for men. Women are less likely to be referred than men are, in particular for angina pectoris. PMID- 10824340 TI - Research capacity in UK primary care. AB - BACKGROUND: Moves towards a 'primary care-led' National Health Service (NHS) and towards evidence-based care have focused attention upon the need for evaluative research relating to the structure, delivery, and outcome of primary health care in the United Kingdom (UK). This paper describes work carried out to inform the Department of Health Committee on Research and Development (R&D) in Primary Care (Mant Committee). AIM: To describe the extent and nature of current research capacity in primary care in the UK and to identify future needs and priorities. METHOD: Funding data were requested from NHS National Programmes, NHS Executive Regional Offices, the Department of Health (DoH), Scottish Office, Medical Research Council, and some charities. A postal survey was sent to relevant academic departments, and appropriate academic journals were reviewed from 1992 to 1996. In addition, interviews were conducted with academic and professional leaders in primary care. RESULTS: Overall, total annual primary care R&D spend by the NHS and the DoH was found to be 7% of the total spend, although annual primary care R&D spend differs according to funding source. Journals relating to primary care do not, with some notable exceptions (e.g. British Journal of General Practice, Family Practice), have high academic status, and research into primary care by academic departments is, with perhaps the exception of general practice, on a small scale. The research base of most primary care professions is minimal, and significant barriers were identified that will need addressing if research capacity is to be expanded. CONCLUSION: There are strong arguments for the development of primary care research in a 'primary care-led' NHS in the UK. However, dashes for growth or attempts to expand capacity from the present infrastructure must be avoided in favour of endeavours to foster a sustainable, long-term research infrastructure capable of responding meaningfully to identified needs. PMID- 10824341 TI - The WOMB (Women's views of birth) antenatal satisfaction questionnaire: development, dimensions, internal reliability, and validity. AB - BACKGROUND: Antenatal services continue to change, stimulated by the Changing Childbirth report. Women's views should be an important component of assessing the quality of such services. To date, no published quantitative multidimensional assessment instrument has been available to measure their satisfaction with care. AIM: To develop a valid, reliable, multidimensional questionnaire to assess quality of antenatal care. METHOD: A multidimensional satisfaction questionnaire was developed using psychometric methods. Following fieldwork to pilot a questionnaire, three successive versions of it were given by midwives to pregnant women in their final trimester in nine trusts in the old South Western region of England. Their replies were analysed by principal components analysis (PCA) with varimax rotation; internal reliability was assessed by Cronbach's alpha. Face, content, and construct validity were all assessed during development. RESULTS: Out of 196 women, 134 (68.4%) returned the pilot questionnaires. One hundred and seventy-two (57.3%) out of 300 women returned version 1 of the WOMB (WOMen's views of Birth) antenatal satisfaction questionnaire proper, 283 (56.6%) out of 500 returned version 2, and 328 (65.6%) out of 500 returned the final development version. This final version consisted of 11 dimensions in addition to a general satisfaction one. These were [Cronbach's alpha]: five related to antenatal clinic characteristics (travelling to clinic [0.75], waiting at clinic [0.90], clinic environment [0.69], timing of appointment [0.78], car parking [0.85]), three 'professional' characteristics (professional competence [0.80], knowing carers [0.79], information provided [0.81]), antenatal classes [0.76], social support from other pregnant women [0.83], checking for the baby's heart beat [0.63]. There were significant moderate correlations (range = 0.24 to 0.77) between individual dimensions and the general satisfaction dimension. Women's dimension scores were significantly related to age, parity, social class, and best educational achievement. CONCLUSION: This multidimensional satisfaction instrument has good face, content, and construct validity, and excellent internal reliability. It could be used to generally assess antenatal services or to screen them to detect areas where further in-depth qualitative enquiry is merited. Its sensitivity to change over time, external reliability, and transferability to non Caucasian groups needs to be assessed. PMID- 10824342 TI - Stopping drug treatment of hypertension: experience in 18 British general practices. AB - BACKGROUND: Of the many reports published describing the effect of withdrawing antihypertensive medication from patients who have well-controlled blood pressure, none have been major British general practice studies. Studies from other settings have shown that a substantial minority can do so without harm or resulting in the relapse of their hypertension. AIM: To determine the proportion of hypertensive patients who could have their medication withdrawn without relapse, and to seek factors associated with success at withdrawal. METHOD: A longitudinal observational study in 18 general practices in north-east England. Practices selected and managed patients to guidelines suggested by the study protocol. Data were abstracted from records by practice staff over three years of follow-up. RESULTS: A total of 196 out of 224 (88%) patients were followed up. Forty-three (22%) of these 196 remained normotensive off medication for the whole study. A total of 108 (55%) of the 196 had restarted medication by three months. Twenty-six (31%) of the 84 males, but only 17 (15%) of the 112 females, remained off medication. No differences in age, morbidity, symptoms, or biochemical parameters occurred between the group who stayed off medication and those who restarted it. Apart from male sex, no factors were found that enabled the prediction of patients more likely to succeed at stopping medication. CONCLUSIONS: One-fifth of well-controlled hypertensives in British primary health care could have their medication withdrawn without the relapse of their hypertension or any harm. Of those that do relapse, over half are likely to have done so before three months. Life-long observation of all patients is essential. PMID- 10824343 TI - Are child health surveillance reviews just routine examinations of normal children? AB - BACKGROUND: The provision and content of child health surveillance (CHS) has changed greatly since 1990. However, its value continues to be questioned. The introduction of the personal child health record (PCHR) has provided a new means of collecting data about CHS. AIM: To identify what problems are recorded at CHS reviews in the PCHR during the first year of life, and what follow-up/referrals result directly from these reviews. METHOD: A total of 28 practices were recruited from one health authority. All babies born to mothers registered with study practices during one year were followed up. Health visitors returned copies of CHS reviews recorded in children's PCHRs. Written comments on returned reviews were analysed. RESULTS: In all, 2308 babies were entered into the study and 2001 (87%) were followed up for one year. A total of 7848 (78%) CHS reviews were returned. Physical problems were recorded in 58% of children at the 10-14 day, 35% at the six to eight week, and 39% at the six to nine month review. Of physical problems recorded at CHS reviews, 30% required follow-up in primary care and 7% required referral to hospital. Other problems were recorded less frequently and health promotion was recorded at only 7.5% of CHS reviews. CONCLUSION: Child health surveillance provides important opportunities to discuss problems that may cause parental concern and to identify children requiring treatment or follow-up. The design and use of the PCHR needs to change to reflect increasing emphasis on health promotion. PMID- 10824344 TI - How accurately do primary health care professionals use cardiovascular risk tables in the management of hypertension? AB - It has been suggested that use of cardiovascular risk tables in hypertensive patients might improve clinical management. As part of a randomised controlled trial, we evaluated the accuracy of health professionals' use of the New Zealand risk tables in primary care consultations. Chance-corrected agreement between health professionals' assessments of absolute risk was only moderate (weighted kappa = 0.56 at the 12-month follow-up). Inaccurate use of cardiovascular risk tables may be a barrier to effective implementation of research evidence in the management of hypertension. PMID- 10824345 TI - Stopping smoking and body weight in women living in the United Kingdom. AB - Most studies of the effect of stopping smoking on weight in women have been cross sectional in design and have been conducted abroad. A survey of subjects still participating in the Royal College of General Practitioners Oral Contraception Study was used to examine changes in weight among women of different smoking status and living in the United Kingdom. During roughly a 26-year period, there was a threefold increase (to 15% in 1994/1995) in the prevalence of obese (body mass index [BMI] > 30) women among the cohort. Women who stopped smoking had the largest increases in mean BMI and in the proportion of obese women at 1994/95. These results support the notion that stopping smoking leads to weight gain. PMID- 10824346 TI - The prevalence of gender dysphoria in Scotland: a primary care study. AB - A questionnaire was sent to senior partners in all general practices in Scotland designed to elicit experience of patients with gender dysphoria: a subjective experience of incongruity between genital anatomy and gender identity. Responses were received from 73% of practices. The prevalence of gender dysphoria among patients aged over 15 years was calculated as 8.18 per 100,000, with an approximate sex ratio of 4:1 in favour of male-to-female patients. One-third of gender-dysphoric patients known to practices had registered in the preceding 12 months, suggesting that patients with this condition are increasingly likely to present for medical care. PMID- 10824347 TI - The ethnic minority linkworker: a key member of the primary health care team? AB - We present an evaluation of the role of a link-worker trained in health promotion and aspects of chronic disease management. A shift in workload occurred from the practice nurse to link-worker, and there were improvements in asthma and diabetes care. A link-worker can be successfully trained to do traditional nursing tasks, which permits a change of role for the practice nurse and can have a beneficial effect on the processes of chronic disease management. PMID- 10824348 TI - Recommendations for future studies: a systematic review of educational interventions in primary care settings. AB - Systematic reviews are an important part of the current move towards evidence based practice. Independent reviewers use a variety of search strategies to identify and assess relevant articles in the field of concern. Criteria for quality must be agreed and articles evaluated accordingly. This study systematically reviewed educational interventions targeting physicians in primary care (excluding hospital clinic and academic settings) to determine their effectiveness in changing behaviour and to investigate whether studies gave information about the resource implications of the interventions described and their rationale for choosing a particular target group. Studies in English, French, or German language journals were included. The review applied the criteria of the Cochrane Collaboration for methodological quality of studies (but was not conducted under the auspices of the Cochrane Collaboration). The results showed that relatively few studies had occurred in primary care compared with academic and hospital clinic settings. Many articles did not fit the criteria for rigour of method, and those that did were very heterogeneous in method and target group. Only two studies assessed resource implications, and one study also calculated economic benefits. The review suggests that future studies should either target geographical areas or doctors with an identifiable learning need associated with patient outcome, and that studies should be evaluated on their 'intention to educate'. Evaluations of educational initiatives need to describe the resource implications versus measurable benefits of the intervention to make their studies useful to policymakers and planners of educational provision. PMID- 10824349 TI - An information retrieval service to support clinical decision-making at the point of care. AB - The information retrieval systems currently available in general practice, such as Medline, and web search engines are passive and relatively difficult to access during consultations. Emergent technologies, including the National Electronic Library for Health, offer opportunities for more active decision support. We examine the extent to which information retrieval could support primary care consultations by examining the impact of the new technology at different stages of the consultation. We advocate a system whereby professional organisations concerned with quality of care, such as the Royal College of General Practitioners, might contribute the the process. PMID- 10824350 TI - Methodological flaws exist in the study of patient-centredness of consultations. PMID- 10824351 TI - Time to go public on performance. PMID- 10824352 TI - Pharmacist intervention in general practice. PMID- 10824353 TI - Implications of the UKPDS Study for general practice care of type 2 diabetes. PMID- 10824354 TI - Use of HRT in an inner-city general practice. PMID- 10824355 TI - Small groups and preparation for FBA. PMID- 10824356 TI - Not poetry, please... PMID- 10824357 TI - Cardiac rehabilitation: exercising the heart. PMID- 10824358 TI - Does diet have a role in the treatment of hyperlipidaemia? PMID- 10824359 TI - Chest pain: is the history useful? PMID- 10824360 TI - Complications of a simple procedure: de Quervain's disease revisited. AB - We reviewed the results of 22 operations performed on 21 patients for the relief of de Quervain's disease over the last six years. At a mean follow-up of 34 months (range 4-78 months), 18 of 22 wrists had complete relief of their original symptoms. One patient required reoperation because of inadequate decompression, and two others await further surgery. Most operations (14 out of 22) were performed under local anaesthesia, and 17 out of 22 used a longitudinal incision. The use of a longitudinal incision was associated with a significant risk of complications: four patients had a poor cosmetic result, and six showed evidence of superficial radial nerve injury. Two patients had a wound infection, and one developed reflex sympathetic dystrophy. Although surgical decompression for de Quervain's disease is effective in curing the symptoms in most patients, using a longitudinal incision is associated with poor wound healing and damage to the terminal branches of the radial nerve. PMID- 10824362 TI - The effects of Valette on skin and hair: a post-marketing surveillance study. AB - The effects of Valette--an oral contraceptive containing ethinyloestradiol 0.03 mg and the antiandrogenic progestogen dienogest 2.0 mg--on the skin and hair were surveyed over 63,474 cycles in 10,718 women in routine gynaecological practice. Improvements were greatest in women with severe or moderate androgen-related symptoms. After six cycles, < 1% of women had severely greasy hair and 6% had moderate greasiness, compared with 11% and 27% at baseline; fewer hair washes were needed per week. The incidence of severe and moderately greasy skin disorders fell from 16% to < 1%, and from 39% to 7.5%, respectively. Self assessments indicated less greasy hair and improved greasy skin disorders in 70% and 81% of women, respectively. The overall effect of Valette on the skin and hair was rated very good or good by 87.5% of women. These results confirm previous observations of a beneficial effect of Valette on androgen-related skin and hair conditions. PMID- 10824361 TI - A randomised comparison of simvastatin versus simvastatin and low cholesterol diet in the treatment of hypercholesterolaemia. AB - There is little evidence to show that strict dietary modification alone confers any significant impact on cardiac events in primary and secondary prevention of coronary heart disease. Given the efficacy of the statins, the need for strict dietary modification in patients on statin therapy has been questioned. This study was performed to assess 1) the added benefit on serum lipid levels of a strict low-fat dietary regimen in patients with hypercholesterolaemia already treated with simvastatin; 2) the efficacy of simvastatin on the lipid profile of our sample Asian population; and 3) the tolerability and side-effect profile of simvastatin. This study was a prospective evaluation of 60 patients with hypercholesterolaemia treated with simvastatin who were subjected to either a normal diet or a dietitian guided low-fat diet. Assessment of the effects on serum lipid levels, side-effects, biochemical and haematological markers were performed. After 24 weeks of treatment, a strict dietitian guided low-fat diet conferred no additional benefit over and above what was achieved by simvastatin alone. Furthermore, a higher dose of simvastatin was needed in the dietitian guided diet group to achieve the target LDL-cholesterol level. Simvastatin resulted in a significant positive alteration of lipid profiles in all patients. The drug was well tolerated, with no significant change in either haematological or biochemical indices. Simvastatin is a highly effective cholesterol-lowering drug with a beneficial effect on the entire lipid spectrum in a cross-section of Asian patients, and is well tolerated. A dietitian guided dietary approach confers no additional advantage once statin therapy has been initiated. PMID- 10824363 TI - Cardiac rehabilitation: from Hellerstein to the millennium. AB - In the 1950s, Herman Hellerstein set out his ideas for the comprehensive rehabilitation of patients recovering from acute cardiac events, using a multidisciplinary team. His approach has been adopted by most developed countries, with varying speeds and enthusiasm. His original ideas have hardly been improved on. However, changes in the demographics and epidemiology of heart disease have led to changes in the patient groups included in cardiac rehabilitation today and, despite falling death rates from coronary disease, the numbers who might benefit from this treatment have grown steadily. Unfortunately, only a minority of patients with heart disease who would benefit are offered rehabilitation. The provision of cardiac rehabilitation in most countries is seriously deficient, and suggestions are made for improving this situation to fulfil Hellerstein's vision. PMID- 10824364 TI - Carcinoma of the breast and hormone replacement therapy for osteoporosis. AB - The benefits of hormone replacement therapy for postmenopausal women, especially in the prevention and treatment of osteoporosis, are well known. It is still not clear whether oestrogen replacement therapy can be given safely to women with breast cancer. The incidence and survival of breast cancer is rising in the UK and increasing numbers of young women with breast cancer treated systemically experience an early menopause and are at prolonged risk of osteoporosis. This review discusses the risk/benefit analysis of oestrogen replacement therapy for breast cancer patients, and alternative therapies: SERMs, phyto-oestrogens and bisphosphonates. A schedule for monitoring osteoporosis for breast cancer patients with a therapeutic early menopause is suggested. PMID- 10824365 TI - Does combination therapy with a calcium channel blocker and an ACE inhibitor have additive effects on blood pressure reduction? AB - Reports in the medical literature have suggested that the effects on blood pressure (BP) of calcium channel blockers (CCBs) and angiotensin-converting enzyme (ACE) inhibitors are additive. Most reports have provided neither a definition of 'additive' nor the necessary information to determine whether the effects are additive. In this review of the medical literature, the effects of combination therapy were defined as additive if the sum of the mean reductions in BP following monotherapy with a CCB and an ACE inhibitor was not significantly different from the mean reduction in BP of combination therapy. The review generally showed that combination therapy is more effective than treatment with either monotherapy alone, based on mean decreases in either diastolic or systolic BP. The studies provided no clear evidence that the effects of combination therapy were either additive or less than additive. PMID- 10824366 TI - Chest pain: panic attack or heart attack? AB - Chest pain is a common presentation to both primary and secondary care physicians but is often non-cardiac in origin. Patients presenting with chest pain may be experiencing a panic attack. Panic disorder is a disabling psychiatric condition with serious consequences, such as impaired social functioning and increased risk of suicide. Comorbidity of panic disorder with other psychiatric conditions is common and often leads to increased severity of anxiety symptoms and a poorer prognosis. The cost of misdiagnosing non-cardiac chest pain is high. It is important for physicians to be able to recognise panic attacks and to distinguish them from cardiac disease, thus avoiding unnecessary use of healthcare resources. This review discusses the prevalence and diagnosis of panic attack and panic disorder in patients presenting with chest pain to primary care physicians and cardiologists. Treatment options for panic disorder are considered, particularly the selective serotonin reuptake inhibitors, which are emerging as the first-line choice for the treatment of panic disorder. PMID- 10824367 TI - Acute pulmonary oedema following smoke inhalation. AB - A case of acute pulmonary oedema after smoke inhalation from a chip pan fire is presented. The role of bronchial and pulmonary circulation in the development of pulmonary oedema after smoke inhalation is discussed. We stress the importance not only of observation after smoke inhalation, as the manifestation of pulmonary oedema may be delayed, but also of a baseline chest X-ray before admission for comparison. PMID- 10824368 TI - Stigma and Alzheimer's disease: causes, consequences and a constructive approach. AB - The term 'Alzheimer's disease' has entered the vocabulary of ordinary people. This has been useful, encouraging patients and families to seek medical help for conditions that were previously neglected as inevitable consequences of old age. Yet Alzheimer's itself can carry negative connotations. Some people who believe they have Alzheimer's disease are suffering from less sinister and more easily treated conditions. Even when the diagnosis is confirmed, many months of happy and worthwhile life continue for most patients and families if they are given appropriate information and support. It is important that neither families nor clinicians see life with presumed Alzheimer's disease as of no value. PMID- 10824369 TI - The role of tirofiban in acute coronary syndromes. AB - Platelet activation and aggregation play an important and essential role in the formation of intracoronary thrombus in acute coronary syndromes (ACS). ACS still carries unacceptably high rates of morbidity and mortality despite intensive antianginal therapy and the wide use of aspirin and heparin. Two glycoprotein IIb/IIIa receptor inhibitors are now licensed for concomitant use with heparin and aspirin in ACS. Glycoprotein IIb/IIIa receptor inhibitors block the final step for platelet aggregation and fibrinogen binding, thus preventing thrombus formation. Tirofiban is a potent, synthetic, non-peptide and specific glycoprotein IIb/IIIa receptor inhibitor. In three major international trials involving over 7200 patients (PRISM, PRISM-PLUS and RESTORE), tirofiban was shown to be well tolerated and to reduce the risk of ischaemic complications in patients with unstable angina, non-Q-wave myocardial infarction and high-risk patients undergoing revascularisation when used in combination with aspirin and heparin. These and ongoing studies are discussed. PMID- 10824370 TI - Mistaken first impressions: a response. AB - In this journal, Cummins et al reported an experiment that examined clinicians' predictions of causes of death from subjects' pre-morbid photographs. They found that clinicians performed at chance. The validity of two of their other conclusions are challenged. First, that clinicians routinely form impressions of how a person will die from their appearance. Second, that certain health professionals use common judgements to form these impressions. These claims are found to be based upon non-falsifiable hypothesis and inappropriate statistics; hence, an experiment employing pictures of feet (or any arbitrary personal detail) would have led the authors to the same conclusions. PMID- 10824371 TI - Perforation of uterus by the GyneFix intrauterine contraceptive device. AB - A case of intrauterine perforation by a GyneFix contraceptive device in a 29-year old parous woman is described. It is the first report of uterine perforation by the frameless, anchored GyneFix in the UK since its introduction here in 1997. Perforation was diagnosed 12 days after insertion and was complicated by visceral perforation and infection. PMID- 10824372 TI - Duodenal obstruction due to a swallowed nasogastric tube. AB - Nasogastric tubes are commonly used, not only in surgical practice but in all disciplines. Their use, however, is not without complications. We present the first reported case of duodenal obstruction due to a swallowed nasogastric tube, and recommend that nasogastric tubes be used in their entirety (uncut) with the splayed distal end intact. This simple procedure will prevent a rare but distressing complication that might be amenable only to laparotomy. PMID- 10824373 TI - An evaluation of an intelligent home monitoring system. AB - A trial was performed of an intelligent monitoring system which used sensors in the home to identify emergencies by detecting deviations from normal activity patterns. The field trial lasted three months. Twenty-two elderly people agreed to participate. Their ages ranged from early 60s to over 85, with two-thirds in the age range 75-84 years. They lived in four different localities within the UK- Ipswich, Northumberland, Merseyside and Nottingham. A total of 61 alerts was recorded, at a mean frequency about one alert per month per client. Of the 61 alerts generated, 46 were classified as false alerts and the other 15 as genuine, although no real emergencies occurred during the study. Many people in the field trial reported enhanced feelings of safety and security, which could help to stimulate independence and help them to remain living in their own homes. The monitoring system increased the care choices available to elderly people and supported and enhanced the carer's role. PMID- 10824374 TI - A randomized, controlled trial of child psychiatric assessments conducted using videoconferencing. AB - We used a PC-based videoconferencing system to conduct child psychiatry assessments. The telecommunications link was six digital lines, giving a total bandwidth of 336 kbit/s. Twenty-three patients (aged 4-16 years), accompanied by their parents, completed two psychiatric assessments, one via videoconferencing and another face to face (FTF). The order of assessments was randomized. Questionnaires were used to record the diagnosis, treatment recommendations and the psychiatrists', patients' and their parents' satisfaction with each assessment. An independent evaluator concluded that in 22 cases (96%) the diagnosis and treatment recommendations made via the videoconferencing system were the same as those made FTF. The psychiatrists stated that videoconferencing assessments were an adequate alternative to FTF assessments and did not interfere with diagnosis. However, the responses from the psychiatrist satisfaction questionnaire showed that they preferred FTF assessments. No significant difference was found in the patients' or parents' satisfaction responses after the two types of assessment. The majority of children (82%) 'liked' using the telepsychiatry system and six (26%) preferred it to a FTF assessment. Most parents (91%) indicated that they would prefer to use the videoconferencing system than to travel a long distance to see a psychiatrist in person. PMID- 10824375 TI - The reliability and acceptability of telemedicine for patients with schizophrenia in Korea. AB - We conducted a pilot study to evaluate telemedicine for patients with schizophrenia. The telemedicine system was connected over the ordinary telephone network at 33 kbit/s. A computer-based patient record was used to view patient summaries and to allow nursing notes to be entered at the patient's home. Fifteen patients with schizophrenia were assessed over the telemedicine system and 15 patients were assessed face to face, using the Brief Psychiatric Rating Scale (BPRS). Our low-bandwidth telemedicine system appeared to be as reliable as higher-bandwidth ISDN systems. In addition, the patients' acceptance of the telemedicine interview, in terms of comfort, ease of self-expression, quality of interpersonal relationship and usefulness, was good in most cases. The only factors significantly affecting the patients' level of acceptance of their particular type of interview were the assessment type (i.e. whether the patient had had a telemedicine assessment or not) and their BPRS score. Since the system was of low cost and was easy to interface with a notebook computer, it could be used support other home-health nursing services. PMID- 10824376 TI - Development of a web-based picture archiving and communication system using satellite data communication. AB - Using the JAVA language we have developed a Web-based picture archiving and communication system (PACS) which allows a remote hospital to access medical images. An asymmetric satellite data communication system (ASDCS) provided a receive-only link for data delivery and a conventional terrestrial link (which could be the conventional telephone network) allowed data transmission. The satellite communication link was 10-30 times faster than the conventional terrestrial link. To increase image transmission speeds over the Internet connection, JPEG and wavelet compression methods were used. The resulting images were evaluated quantitatively by measuring the peak signal:noise ratio and qualitatively by radiologists. Compression ratios of 10:1 or less were deemed acceptable for diagnostic purposes. The system appears to be suitable for teleradiology and telemedicine. PMID- 10824377 TI - Patient cost-benefits of realtime teledermatology--a comparison of data from Northern Ireland and New Zealand. AB - As part of a randomized controlled trial of the costs and benefits of realtime teledermatology in comparison with conventional face-to-face appointments, patients were asked to complete a questionnaire at the end of their consultation. One hundred and nine patients took part in an initial teledermatology consultation and 94 in a face-to-face consultation. The proportion of patients followed up by the dermatologist was almost the same after teledermatology (24%) as after a hospital appointment (26%) and for similar reasons. Two hundred and three questionnaires were completed after the first visit and a further 20 after subsequent visits. Patients seen by teledermatology at their own health centre travelled an average of 12 km, whereas those who attended a conventional clinic travelled an average of 271 km. The telemedicine group spent an average of 51 min attending the appointment compared with 4.3 h for those seen at the hospital. The results of the present study, as in a similar study conducted in Northern Ireland, show that the economic benefits of teledermatology favour the patient rather than the health-care system. PMID- 10824378 TI - A feasibility study of realtime teledermatology in Finland. AB - We conducted a feasibility study of teleconsultation in dermatology using low cost equipment. Patients and their general practitioners took part in consultations from the Primary Health Care Centre in Ikaalinen with a dermatologist 55 km away at the Tampere University Hospital (TAUH). Consultations were performed using standard commercial videoconferencing equipment, a modified document camera and a dermatoscope. A single ISDN line (128 kbit/s) was used for the connection. During the eight months of the study, 25 patients participated in a teledermatology consultation. Their mean age was 45 years (range 4-92). The average time the patient spent in travelling to the videoconsultation (i.e. one way) was 24 min (range 5-65 min). The mean time spent in the teleconsultation was 15 min (range 5-30 min). After the teleconsultation, patients' treatments changed in 19 cases (76%), diagnoses were changed in 13 cases (52%) and 18 patients (72%) did not need to go to the TAUH. The equipment was generally reliable and easy to use. However, the dermatoscope was not very useful and only one of the consultations relied mainly on it. The cost of the teleconsultations for the 18 patients who avoided travel to the TAUH was FM18,627. The total costs for the 18 conventional consultations in the TAUH would have been FM18,034. The main economic benefits of the videoconferencing were attributable to the reduced travelling and hospital costs. The economic benefits of medical education were more difficult to quantify. PMID- 10824379 TI - Patients' views about information technology in the treatment of hypertension. AB - We assessed patient's attitudes to information technology (IT), their experiences of IT, and their attitudes and expectations concerning its use in the treatment of hypertension. Twenty-one patients with high blood pressure were interviewed before they took part in a pilot study of a computer-based intervention to control high blood pressure; they were aged between 32 and 63 years, and had received medication for one year at most, or no medication at all. Content analysis of the interview transcripts revealed six factors that affected the acceptance and use of IT. These were: the patients' general attitude to IT and its use, their experiences of using IT, their perceptions of the barriers to the use of IT, the advantages of using IT in the treatment of hypertension and the usefulness of IT, and their expectations of the computer program. The patients wanted to take part in the intervention and their attitudes were notably positive even when they had little or no experience of using IT. PMID- 10824380 TI - An experimental teleradiology transmission system using a high-speed ATM backbone network. AB - We evaluated the performance of an experimental teleradiology system based on a high-speed ATM backbone network. Image acquisition, transmission and the disk-to display processing times were measured. Computerized tomography (CT) scans printed on 14 inch x 17 inch (36 cm x 43 cm) films were digitized and transferred over the network. The average time for the entire process was 1 min 30 s. Three radiologists interpreted 20 cases. For CT image interpretation, the reading time for one case ranged from 2 to 12 min (mean 6 min 46 s) on a monitor, and from 1 to 3 min (mean 1 min 31 s) with the original film. The ATM backbone network operating at 156 Mbit/s provided sufficient speed for remote consultation. However, further improvements in the operability of the system, especially the image viewing station, are necessary before it will be satisfactory for clinical use. PMID- 10824381 TI - Theory of telemedicine: the expressivity of the virtual patient. PMID- 10824382 TI - Telemedicine in the twenty-first century--report of a workshop. PMID- 10824383 TI - Telemedicine and decision support in emergency ambulances in Uppsala. AB - In Sweden, as in many other countries, paramedics or nurses constitute the majority of prehospital personnel. If tasks usually performed by doctors are to be performed by these personnel, there is a need for guidelines and triage in the handling of medical emergencies. We describe an information management system used in ambulances for data communication, documentation, triaging and presentation of checklists. In most cases, data are input while the patient is being cared for. The information is collected and stored together with data automatically received from the dispatch centre. The latter date are transferred by a mobile radio network to the ambulance. Medical date (e.g. electrocardiograms) are transferred from the ambulance over the data network to the receiving medical facility. All transferred data are finally collected in a database for statistical analysis and follow-up. PMID- 10824384 TI - 'Smart' homes and telecare for independent living. AB - Telecare services and 'smart' homes share a common technological base in information technology and telecommunications. There is growing interest in both telecare services and smart homes, although they have been studied in isolation. Telecare has been driven largely by perceived cost savings and improved service delivery to the home, leading to improved quality of life and independent living. Smart homes are also expected to provide better and safer living conditions. The integration of the two should produce more secure and autonomous living. There are different forms of telecare services, as there are different types of smart homes, each ranging from basic systems involving the use of alarms and the ordinary telephone to intelligent monitoring with sensors and interactive communication. The introduction of these systems has policy implications, such as the need for coordination between health, social services and housing policy makers, which will reduce duplication and inefficient allocation of resources. Successful delivery of telecare to the home is as much dependent on the construction and condition of the housing stock as it is on the ability of the care provider to meet users' needs. If the UK National Health Service (NHS) could replace a significant proportion of domiciliary nursing visits by telephone calls, then savings of up of 200 million Pounds per annum would be possible. PMID- 10824385 TI - Internet-mediated, protocol-driven treatment of psychological dysfunction. AB - An Internet system was developed for assessing psychopathology, for on-line, protocol-driven cognitive-behavioural psychological treatment and for measuring the effects of treatment. The system focused on the treatment of post-traumatic stress. The treatment comprised 10 writing sessions (45 min each) over five weeks. Participants were assessed on-line before treatment, after treatment and after six-week follow-up. After treatment the participants had improved significantly in terms of post-traumatic stress symptoms (P < 0.005) and general psychological functioning (P < 0.005), and this was sustained during the follow up. Nineteen of the 20 participants were clinically recovered after treatment. Reduction in post-traumatic stress symptoms compared favourably to changes in control and experimental groups in trials of similar but face-to-face treatment. PMID- 10824386 TI - Self-treatment of obsessive-compulsive disorder guided by manual and computer conducted telephone interview. AB - While on a waiting list for treatment by therapist-guided exposure and ritual prevention (ERP), patients with obsessive-compulsive disorder (OCD) did self treatment at home guided by a manual plus a computer-driven telephone interview system (BT STEPS). Of 21 patients who used the system for at least three weeks while on the waiting list, one improved so much that subsequent therapist-guided ERP was unnecessary. Progress of the rest with the system predicted later progress with therapist-guided ERP. Improvement after using the system was similar to that of 20 matched historical controls who had had therapist-guided ERP without the prior use of BT STEPS. Outpatient users of BT STEPS needed less subsequent clinician-guided time than did their matched controls. In this pilot study, patients with OCD improved nearly as much with home self-treatment guided by a manual plus computer, as with treatment guided by a behaviour therapist. PMID- 10824387 TI - The value of the World Wide Web for tele-education in radiology. AB - We assessed Websites for radiological education. Of several hundred sites identified, 30 were randomly selected for evaluation by two groups of observers. All Web pages were rated with respect to time required to access the information; structure and organization of the site; image quality; and information content and relevancy. Rapid access was gained to many educational radiology Websites, which contained large numbers of interesting radiological images of good quality. A weak point was the structure and organization of the sites. The greatest variability in ratings was found in the information content and relevancy of the Websites. PMID- 10824388 TI - Telemedicine activity at a Canadian university medical school and its teaching hospitals. AB - Dalhousie University Medical School and its teaching hospitals have been providing clinical telemedicine services since 1987. The object of the present study was to assess the extent and growth of telemedicine at the medical school and teaching hospitals, as well as to evaluate the obstacles to its deployment. This was achieved by conducting structured personal interviews with telemedicine providers. Twenty telemedicine programmes were identified, of which 15 were operational and five were being planned. The number of established telemedicine projects had doubled in the six months preceding the study. A wide variety of telemedicine services were provided, ranging from clinical consultations in a number of medical specialties to patient education, grand rounds and continuing medical education. These services were provided to sites in a wide area in the Maritime region and internationally. The three most important obstacles to the implementation of telemedicine were a lack of knowledge about telemedicine (80% of respondents), time constraints (75%) and funding (70%). PMID- 10824389 TI - Patients' perceptions of a telemedicine specialty clinic. AB - We studied patients' perceptions of telemedicine (interactive video) as a means of delivering specialist oncology/haematology care. Semistructured telephone interviews were performed with 22 patients attending a tele-oncology/haematology clinic in Kansas. Interviews were audio-taped and thematic content analysis of the transcripts was done. The response rate was 96%. All participants expressed satisfaction with their tele-consultations. For 50% of respondents, satisfaction with the tele-oncology/haematology clinic was qualified by two factors: participants also saw the specialist in person on occasions and the clinic was perceived as providing mainly a 'monitoring' function. In addition, 9 of the 22 patients expressed concern about the role of the nurse as a proxy for the doctor in performing certain parts of the physical examination. Overall, participants valued improved access to specialist services but had clear views as to the limitations of such a service. As the use of telecommunications technology grows, patient perspectives merit greater attention. PMID- 10824390 TI - International transmission of tele-ophthalmology images. AB - Tele-ophthalmic consultations were conducted between a hospital in East Java, Indonesia, and an ophthalmology centre in Perth, Western Australia. Twenty-two eyes of 14 subjects were screened for glaucoma using a hand-held fundus camera. Optic disc images comprised 267 x 234 pixels at 24 bit/pixel (187 kByte). The images were compressed and stored together with patient information on a laptop computer. The images were then transmitted to Perth using either a mobile phone or a satellite phone, taking 170 s or 240 s, respectively. Images were also compressed to five different compression levels before transmission. At a compression ratio of 1:5, the images were 36 kByte in size and took 29 s to transmit by mobile phone and 60 s by satellite phone. To measure the loss of quality, the root mean square error was calculated for each colour component, comparing the transmitted and original images. The coefficients of variation were 10% (green), 15% (blue) and 22% (red). The images received in Perth were considered to be of excellent quality and readily interpreted by ophthalmologists in terms of the likely presence of glaucoma. PMID- 10824391 TI - Initial experience with a wireless personal digital assistant as a teleradiology terminal for reporting emergency computerized tomography scans. AB - A new type of terminal device, a wireless personal digital assistant (PDA) based on a GSM digital cellular phone, was used to transmit computerized tomography scans of 21 patients to a neuroradiologist. All transmitted images were suitable for a preliminary consultation and in one case a final report could be made. In 18 cases the findings were compatible with the reference film reading performed later and in three cases there were minor differences of no clinical importance. Transmission of a single image lasted 1 min 30 s and the transmission of a complete brain scan (14 images) took on average 21 min. The total process of transmission and interpretation of a brain examination series took on average 40 min. In this pilot study the neuroradiologist gained essential information in 24% of the cases and beneficial information in 62%. The neuroradiologist considered that the image consultation saved a hospital visit in 15 cases (71%). Although PDA technology is at an early stage of development and has numerous limitations, it is likely that future technical improvements will allow easier clinical consultations for neurosurgeons and neurologists. PMID- 10824392 TI - General practitioners' perceptions of store-and-forward teledermatology. AB - We studied the views of 26 general practitioners (GPs) towards store-and-forward tele-dermatology before its introduction into their practices. A postal questionnaire was developed using Likert-type questions with respondents able to explain their answers in free text. Questions related to the GPs' knowledge, perceptions and expectations of tele-dermatology, as well as their attitudes towards being part of a research trial. Most of the GPs had limited prior knowledge of tele-dermatology. They perceived its role to relate to quicker access to specialist opinions, decreased referrals, increased convenience for patients, diagnosis, and education and teaching. There was an overwhelming view that any system needed to be quick, easy to use, efficient and reliable. Concerns were expressed about being part of the clinical trial, using new technology and an increased workload. The future of tele-dermatology was thought to depend on the clinical adequacy of the system. PMID- 10824393 TI - Supervision of thrombolysis of acute myocardial infarction using telemedicine. AB - The treatment of acute myocardial infarction (MI) constitutes a significant problem in remote geographical areas of Greece. Furthermore, thrombolysis, the treatment of choice in the early phase of acute MI, requires the supervision of an expert. We have used thrombolytic treatment, using telemedicine, in remote medical centres. The Onassis Cardiac Surgery Centre was linked to six remote Aegean islands via telemedicine systems which permitted the transmission of 12 lead electrocardiograms (ECGs). The thrombolytic agent anistreplase was administered to patients with acute MI. Supervision, including consultation for treatment of complications, was achieved using the telemedicine system. One hundred and fifty-two ECGs were transmitted during 24 months, of which 108 (71%) indicated specific treatment of a cardiac condition. Ten cases were diagnosed as having acute MI and eight of these were treated with anistreplase. All patients survived acute MI and complications were treated locally. The application of thrombolytic treatment in acute MI is feasible in remote areas, with the use of a telemedicine system. PMID- 10824394 TI - Renal telemedicine to the home. PMID- 10824395 TI - [Recent trends of allergen specific hyposensitization therapy]. PMID- 10824396 TI - [Growth factor, growth factor receptor and transcription factor for differentiation of mast cells]. PMID- 10824397 TI - [Summary of therapeutic guideline for atopic dermatitis]. PMID- 10824398 TI - [Four cases with latex allergy followed by anaphylaxis to chestnut]. AB - It is well known that patients with latex allergy have cross-reactions to various fruits, which is called a latex fruit syndrome. We report four cases with latex allergy followed by anaphylaxis to chestnut. They are all nurses of our hospital, who has personal history of atopic diseases. There were varieties in the methods of processing chestnut, presence of epicutaneous contact to chestnut, and clinical courses among the cases. All cases had positive skin prick test reactions while only two cases showed specific IgEs measured with AlaSTAT to chestnut. This fact suggests that we have to warn the risk of anaphylaxis even if one had not shown a serum specific IgE. We could follow the clinical courses and study specific IgEs to chestnut and latex in the two cases for more than two years. The titer of specific IgE was increased in the one, who could not avoid eating chestnut and contact to latex, while it was decreased in the other who could avoid the exposure to the antigens. Hevein is one of the panallergens among latex and related fruits. We studied specific IgEs to hevein on these four cases and 12 normal controls. The results showed that the former had significantly higher values of sIgEs to hevein compared to the latter (p < 0.05). We conclude that a patient with latex allergy has a high risk of contact urticaria or even anaphylaxis to the related fruits such as chestnut so that we recommend the patient with latex allergy to avoid them. PMID- 10824399 TI - [Evaluation of MAST-26, newly developed system for the detection of IgE antibodies]. AB - MAST-26 is a new allergy testing system which allows simultaneous determination of allergen-specific IgE antibody levels for 26 allergens using 200 microliters of patient serum. To evaluate the effectiveness of MAST-26 for the detection of allergen-specific IgE antibody, a total of 100 serum samples were collected from allergic patients at five facilities, and allergen-specific IgE antibody was measured by MAST-26. 14% of them were positive to > or = 10 allergens. If the patients had severe allergic symptoms, they were likely to show positive to many allergens. A good correlation was found between the results obtained by MAST-26 with those measured by CAP RAST and by intradermal skin test. It was concluded that MAST-26 is an effective screening method for the detection of allergen specific IgE. PMID- 10824400 TI - [Effect of beclomethasone increment on airflow limitation in asthmatic children treated with high dose beclomethasone]. AB - In order to evaluate the effect of higher dose BDP therapy (1200-1660 micrograms/day), we studied 12 asthmatic children (mean age 9.7 years-old) with airflow limitation on respiratory function test who were asymptomatic with high dose BDP therapy (800 micrograms/day). After 4 weeks of higher dose BDP therapy, FVC, FEV1 and V50 were significantly improved, but those improvement were insufficient compared with those after salbutamol inhalation. The personal best values after salbutamol inhalation were not different in every parameter of respiratory function test between BDP 800 micrograms/day and 1200 micrograms/day. We conclude that less than 800 micrograms of daily BDP is generally adequate for prevention in most asthmatic children, because higher dose BDP therapy is no more effective on respiratory function in those treated with 800 micrograms of daily BDP, and that the best value of respiratory function after salbutamol inhalation is not always a goal of high dose BDP therapy. PMID- 10824401 TI - [Multiple nodular and patchy shadows in chest radiograph in a patient with autoimmune hemolytic anemia associated with chronic thyroiditis]. AB - A 65-year-old woman was admitted because of dyspnea and persistent cough. Her chest radiograph and CT films revealed multiple nodular and patchy shadows in bilateral lungs, indicating bronchiolitis obliterans organizing pneumonia (BOOP) like shadows. Autoimmune hemolytic anemia was diagnosed because peripheral blood cell analysis revealed anemia (hemoglobin, 5.5 g/dl) with increased reticulocytes (253/1000), and positive direct and indirect Coomb's tests. In addition, decreased diffusion capacity and mild hypoxemia (PaO2, 71 Torr) were recognized. Bronchoalveolar lavage fluid (BALF) showed the increased number of total cells and lymphocytes, and a decreased CD4(helper)/CD8(suppressor) ratio (= 0.65). Histopathological features of transbronchial lung biopsy showed an infiltration of lymphocytes in the alveolar walls and spaces, and thickening of the alveolar walls. A discrepancy was apparent among roentgenogram, histopathological findings and BALF results. Open lung biopsy had not been performed because of severe hemolytic anemia and lack of informed consent. In addition, the patient suffered from chronic thyroiditis with positive thyroid autoantibody. After oral administration of prednisolone, both pulmonary shadows and anemia improved. We concluded that autoimmune hemolytic anemia may be associated with steroid sensitive interstitial lung lesions in some cases. Further immunologic studies are necessary to elucidate the relationship between autoimmune hemolytic anemia and interstitial lung shadows. PMID- 10824402 TI - [The correlation between the exacerbation of bronchial asthma and picornavirus (human rhino virus) infection in throat gargles by RT-PCR]. AB - Viral infection is one of important factors to cause the exacerbation of bronchial asthma. We have investigated 167 adults of asthmatics to clarify the correlation between viral infection and exacerbation of asthma. Patients were classified to four group by the symptoms of common cold and asthma attack. Furthermore, we have examined Picornavirus and Human rhino virus RNA from throat gargles of patients using RT-PCR (reverse transcription--polymerase chain reaction) method. Forty of 65 (61.5%) asthmatics with common cold revealed asthma attack and common cold was significantly associated with acute exacerbation of asthma (p < 0.01). We identified Picornavirus RNA, which include 113 of Human rhino virus serotypes and enterovirus, from the samples of 16 of 52 (30.8%) patients who had acute exacerbation. It was significantly higher than the detection rate of viral RNA from patient without asthma attack. Furthermore, we analyzed Human rhino virus RNA from the same samples by RT-PCR and 93.7% of Picornavirus were identified as Human rhino virus. Taken together, these findings suggest that common cold is significantly associated with the exacerbation of bronchial asthma. Human rhino virus infection might be one of important virus in this procedure. PMID- 10824403 TI - [False positivity elicited by house dust mite allergen contamination in cat specific IgE antibody determination]. PMID- 10824404 TI - Dynamics of antibodies in Borrelia burgdorferi sensu lato infections. AB - Antibody response in infections with Borrelia burgdorferi sensu lato is generally considered to be slow and delayed, but exact studies concerning this question are hardly found in the scientific literature. During 1994-1998 at least two serum samples were submitted for serological testing from more than 1200 patients. An immunofluorescence test was performed paralelly with two pools of antigen (B. bg.s.s. + B. afzelii, and two serological different strains of B. garinii, all of local origin). In 92-96% of patients no change of antibody level was found in repeated tests, about 20% of them being negative (< 1:512). In 2-4% of cases a significant increase and in less than 1% a decrease or a second wave of the rise of the antibody level was observed. From the 58 patients with increasing antibody levels 9 had ECM (mostly children), 2 cardiovascular, 8 musculo-skeletal and 12 neurological disorders. In the remaining cases the diagnosis was not specified (A 69). Only in 9 cases a rise of the titer appeared during 3 weeks after the first negative sample, at contrary in 7 cases no rise of the titer was seen in that time. 2 patients were still after 1 month, 3 after 3 months and 1 even after 7 months (patient with a positive CSF culture) serologically negative. A 2 titer step decrease was observed in 4 cases during 1 year and once a 3 titer decrease after 4 years. CONCLUSION: A rise of antibodies can be seen in all clinical manifestations of Lyme disease with prevalence in the early types of the disease. The rise of antibodies is often delayed or even inhibited, apparently mostly by an early efficient treatment. Whether a late rise of antibodies or its long persistence indicates a latency of the infection, requires more detailed studies. (Tab. 5, Ref. 6.) PMID- 10824405 TI - Effect of early intervention of high-frequency oscillatory ventilation on the outcome in pediatric acute respiratory distress syndrome. AB - BACKGROUND: Acute respiratory failure in both pediatric and adult patient populations has been extensively studied with recent emphasis on ventilation strategies that can effect mortality outcome. This research in adults has focused on definitive trials of lung protective strategies that have been proposed following preliminary reports of their potential benefits. High frequency oscillatory ventilation has also been described as a lung protective strategy. For many institutions HFOV is today considered a routine therapy as a "rescue" method in acute pediatric respiratory failure. Because HFOV is considered to be a "rescue" therapy, intervention with HFOV is usually in the later stages of acute respiratory failure and consideration of the time to intervention has not been previously examined. OBJECTIVE: To evaluate the effect of time to intervention with high-frequency oscillatory ventilation (HFOV) on the survival of children with severe acute hypoxemic respiratory failure who were managed with lung protective strategies on conventional mechanical ventilation (CMV). METHODS: Twenty-six consecutive patients older 1 month of age with severe hypoxemic respiratory failure and ARDS who at some point in their management were treated with HFOV were evaluated. The mean age was 3.7 years and included three patients treated in the Pediatric Intensive Care Unit (17, 19 and 24 years). Mean weight was 13.8 kg and there were 17 males and 9 females. Nine patients met Extracorporeal Membrane Oxygenation (ECMO) criteria, although only two patients were cannulated. Upon admission to the PICU, patients were initially managed with lung protective strategies using Pressure Controlled Ventilation (PCV) or Pressure Regulated Volume Control (PRVC) modes with limited peak inspiratory pressure, high positive end-expiratory pressure, and permissive hypercapnia. If a Pa-CO2 reached > 75 torr (10.0 kPa) and/or pH < 7.20, tracheal gas insufflation (TGI) was instituted. If FiO2 remained above 0.6 and mean airway pressure (Paw) exceeded 15 cmH2O in order to maintain arterial saturation above 89% or if hypercapnia and/or acidosis on CMV with TGI persisted, the patients were switched to HFOV. An "Optimal Volume Strategy" with HFOV was utilized to recruit alveoli and optimize lung volume. Patients were returned to CMV when their mean airway pressure were between 15 and 20 cmH2O, FiO2 < 0.6, had no evidence of air-leak and/or improved chest X-rays, and did not desaturated during airway suctioning. Patients were offered ECMO if the hypoxemia persisted on HFOV and there were no contraindications to its use. The patients were stratified for analysis by the time to intervention with HFOV. Early intervention was defined as within the first 24 hours of mechanical ventilation (17 patients) and late intervention defined patients beyond 24 hours (9 patients). Demographic data (gender, age, weight, admission PRISM score), time of each mode of ventilation, oxygenation indices and outcomes were recorded for both groups of patients. MAIN RESULTS: The severity of respiratory failure at the time of HFOV intervention was comparable in both early and late groups (PaCO2/FiO2 83 vs. 79 torr, oxygenation index 27 vs. 33, AaDO2 421 torr (56 kPa) vs. 413 torr (55 kPa)). There were no differences in mean age, weight, admission PRISM score length of HFOV, length of CMV after HFOV (CMV post-HFOV) and the total duration of mechanical ventilation between the groups. We found a statistically significant difference in mortality with 58.8% of the early intervention patients surviving while only 12.5% of the late intervention patients survived. The overall survival rate was 42% (11/26 patients). CONCLUSION: Early use of HFOV within the first 24 hours of acute hypoxic respiratory failure in pediatric patients is associated with better survival. Use of this therapy should be considered early in the course of treatment of any pediatric patient meeting this definition. (Tab. 2, Fig. 1, Ref. 28.) PMID- 10824406 TI - [Detection and diagnosis of hereditary monogenic neurologic diseases in Slovakia]. AB - Authors present a clinical symptoms recapitulation of the most important monogenic hereditary neuromuscular diseases, their molecular-genetic causes and the possibilities of diagnostic on the level of DNA analysis. Low detectability of these pathologic states in Slovak republic is stressed and possible causes of this state are analyzed. (Ref. 10.) PMID- 10824407 TI - [Coexistence of cholinergic and nitrergic neurotransmitters in the spinal cord of rabbits]. AB - Nitric oxide (NO) plays a major role as a neuronal messenger molecule. NO has been assumed to act as a retrograde signalling molecule that modulates transmitter release. Acetylcholine (ACh) is known to function as a typical neurotransmitter. In the present work the presence of both transmitters (NO and ACh) and their possible relations in the rabbit spinal cord were examined. In our experiments histochemical methods for the visualisation of acetylcholinesterase (AChE) and NADPH diaphorase (NADPH-d) were used. Both histochemical methods were performed separately and together on the same sections of the thoracic spinal cord. NADPH-d positive dark blue stained neurons were mainly detected in superficial and deep layers of dorsal horn, preganglionic autonomic neurons and pericentral area (1). The presence of AChE positive amber yellow neurons was confirmed mostly in motoneurons located in ventral horns and then in neurons of the intermediate zone. Except for the above mentioned also double-labeled neurons containing both yellow and dark blue histochemical product were noticed. Their presence was confirmed in the intermediate zone and in the pericentral area. Thus, the coexistence of NADPH-d and AChE was confirmed in the area of interneurons. These observations suggest that NO may play a role in the control of cholinergic neuronal activity and that NO can be involved in the modulation of synaptic transmission. (Fig. 9, Ref. 21.) PMID- 10824408 TI - [Frequency of HLA-A, HLA-B and HLA-Cw antigens in the Slovak population]. AB - Results on HLA-A, -B and -Cw antigen frequencies in the Slovak population are presented. HLA-A, -B, -Cw antigens were determined in 654 healthy unrelated individuals. The highest frequency was observed for the antigens HLA-A2, -A1; HLA B12, -B35, and HLA-Cw8. The least frequent antigens were HLA-A34, -A36, HLA-B58, B67, -B70, -B77, and HLA-Cw8. The results were compared with those of the previous study and with those of Czech, Austrian and Hungarian populations. No statistically significant differences were observed. (Tab. 5, Fig. 2, Ref. 9.) PMID- 10824409 TI - [A homozygous quantitative defect of alpha 2-antiplasmin in a family from central Slovakia]. AB - Alpha2-antiplasmin is the main inhibitor of plasma fibrinolytic system. An inborn defect of alpha2-antiplasmin was first described by Koie et al. in 1978 in connection with severe haemorrhage syndrome. The authors present a case report of 45 years old woman living in middle Slovakia with severe haemorrhagic syndrome started in childhood (epistaxis, skin and muscle hematomas, appendectomy with 6 weeks recovery, hardly manageable haemorrhage after teeth extractions, menorrhage, metrorrhage). Laboratory tests were negative for platelet function defects and coagulation system defects. Low level of alpha2-antiplasmin activity (10%) was detected with use of synthetic chromogenic substrate method and low amount (2%) with ELISA method. In asymptomatic daughter was decreased level of alpha2-antiplasmin (activity 51%, quantity 32%) detected. On the basis of patient history, laboratory investigations and comparison with published cases the haemorrhagic syndrome is considered to be an inborn homozygous quantitative defect of alpha2-antiplasmin. Detection of the defect in further haemorrhagic and risk situations (including laparotomy, multiple teeth extractions, obstetric surgery) led to following therapeutic measures: careful local care [by procedures], tranexamic acid in sufficient dose [4 g/day in continual i.v. infusion, or 4 x 1 g in 1/hour infusions, 4 x 1 g perorally], in sufficient duration [14 days by procedures]. Therapeutic approach after detection of the defect is efficient. (Tab. 2, Ref. 13.) PMID- 10824410 TI - [Importance of certain hemostatic indicators as cardiovascular risk factors in type 2 diabetes]. AB - BACKGROUND: Abnormal levels of some hemostatic factors in patients with type 2 diabetes mellitus are associated with increased risk of severe cardiovascular complications. OBJECTIVES: For the purpose of clinical practice it is necessary to specify most relevant coagulation factor abnormalities pertaining to a particular patient's risk profile (e.g. diabetes duration, his/her lipoprotein profile), since a routine examination to their full extent is in common ambulatory practice hardly manageable. This information is also needed for aiming the efforts to influence them purposefully. METHODS: The investigated group included 42 out-patients with type 2 diabetes mellitus. Selected hemostatic factors and lipoproteins were measured with standard methods. Patients were stratified into terciles based on supposedly causative factors (diabetes duration for vWF and lipoproteins for F VII), these relations being also examined in the whole group using regression analysis. MAIN RESULTS AND CONCLUSIONS: vWF (antigen) plasma concentration mean value in the whole group (in % of reference plasma: 149.917; 95% CI 134.296 to 165.538) was on the upper limit of the normal value reference range (56 to 140), exceeding it distinctly in the tercile III with the longest diabetes duration (vWF mean was 180.115; 95% CI 151.961 to 208.270). The mean in this tercile was also by 42.187 (95% CI of diff. 10.231 to 74.142; sign. level 0.012) higher than in the tercile I with the shortest diabetes duration. These 2 variables significantly correlated (r = 0.345; p = 0.025) also in the whole group. F VII plasma concentration mean values in terciles III with highest triacylglycerol (TAG) or total cholesterol (TCH) serum levels (in %: 129.286, 95% CI 121.117 to 137.455 or 121.071, 95% CI 109.228 to 132.915, respectively; the normal value reference range being 70 to 130) clearly surpassed the analogous values in tercile I by 35.0 (95% CI of diff. 22.793 to 47.207; sign. level < 10(-5)) or by 26.0 (95% CI of diff. 10.815 to 41.185; sign. level 0.002), respectively. There were also positive significant correlations between lipoproteins (TAG, TCH or LDL) and F VII in the whole group (r = 0.697, r = 0.580, r = 0.397; p < 0.0001, p < 0.0001, p = 0.018, respectively). Therefore the estimation of vWF seems to be useful in all diabetic patients and most elevated values are to be expected in long-duration-diabetics. F VII should be cared for especially in patients with concomitantly elevated TAG, TCH or LDL cholesterol serum levels. (Tab. 6, Fig. 4, Ref. 24.) PMID- 10824411 TI - [Nutritional risk factors of a vegetarian diet in adult lacto-ovo vegetarians]. AB - Risk nutritional factors of alternative alimentation detected in childhood were evaluated in a group of adult lactoovovegetarians (n = 47). The levels of iron, calcium, zinc, total proteins, gluthatione, plasmatic profile of fatty acids and the lipoperoxidation product in correlation with the values of antioxidative vitamins were studied. The results are compared with mean sample upon mixed nutrition (omnivores n = 42). In both groups were the mean values of iron, calcium and zinc in physiological range, but significantly lower in vegetarians. In the alternative nutrition group was in 21 percent of probands hyposiderinemia detected (vs 5 percent in the omnivores group), in 19 percent of probands hypocalcemia (vs 9 percent) and in 6 percent hypozincemia (vs 0 percent). Full bodied mixture of milk proteins, egg proteins and vegetable sources in lactoovovegetarians ensured sufficient protein saturation and caused significantly higher level of blood gluthatione (intake also in food). Increased value of fatty acid peroxidation index was not due to increased lipoperoxidation in lactoovovegetarians--significantly lower levels of conjugated fatty acid dienes were determined. This was ensured by sufficient protection by means of essential antioxidants--the levels of vitamins E, C, beta-carotene are in vegetarians significantly higher. These are overtreshold values representing reduced risk of free-radical diseases. Lactoovovegetarians had significantly higher content of linoleic and alpha-linoleic acids in plasma. Values of polyunsaturated fatty acids with C20 and C22 and 3-6 double bonds were similar to values in omnivores. In probands on alternative nutrition with iron deficit was significantly lower activity of delta 6 desaturase determined. (Tab. 3, Ref. 45.) PMID- 10824412 TI - [Clinical importance of the Gavornik classification of mushroom poisoning]. AB - The lack of experience and fundamental knowledge about mycology by some mushroomers is one of the leading causes of increasing occurrence of fatal mushroom poisonings. Mushroom intoxications are caused not only by poisonous mushrooms (true primary intoxications), but under certain conditions also by edible mushrooms (secondary intoxications, false intoxications, pseudo intoxications). Apart from fresh mushrooms intoxications may result also from preserved mushrooms (sterilized in pickles, soured, dried, used for preparation of mushroom extracts, powders, etc.), which are used as garnish. (Tab. 1, Ref. 44.) PMID- 10824413 TI - [Immunostimulatory and other beneficial health effects of lactic acid bacteria]. AB - Lactic acid bacteria in functional foods can transiently colonize the intestine of man and exert beneficial probiotic effects. These were observed in a group of adult subjects administered daily by a lyophilized Enterococcus faecium M-74 in the form of waffles (Dr. Ebi) during nine weeks of a double blind placebo controlled clinical trial. The results showed significant immunostimulatory effect on both phagocytosis by neutrophils and antibody production. PMID- 10824414 TI - [Atypical mercury poisoning]. AB - A case report documenting rare intravenous intoxication with metal mercury. Because of early diagnosis and treatment only minimal clinical symptoms were detected in the patient. PMID- 10824415 TI - [Autoimmune hemolytic anemia and chronic giant cell hepatitis]. AB - The authors describe a case report of a six months old boy with autoimmune hemolytic anemia and giant cell hepatitis. This is a very rare syndrome in infancy. After eleven months of intensive immunosuppressive treatment (glucocorticoids, cyclosporin A, azathioprin, intravenous immunoglobulins, cyclophosphamid) no improvement of hemolysis was observed and therefore splenectomy was indicated. 1 year after the treatment was finished the patient is in good general condition and his hematologic and biochemical parameters are physiological. PMID- 10824416 TI - [Total excision of the mesorectum]. AB - In retrospective study authors present early results after the introduction of total mesorectal excision (TME) in rectal cancer surgery. In the period (1995 1998) 50 patients had low anterior resection. Of these 33 were performed with total mesorectal excision. 28 of 33 patients had curative resection, 5 patients had palliative resection due to hepatic metastases. Postoperative mortality rate in TME group was 12% due to cardiac disease. The rate of anastomostic failure was 6%. During minimally 2 years follow-up in 15 patients, local recurrence as well as distant metastases had developed in 1 patient (6.6%) respectively. PMID- 10824417 TI - [New pedagogic methods in anatomy: experience at Cambridge University]. AB - The expansion of knowledge in basic medical sciences is not linked to the time assigned for the teaching of anatomy to medical undergraduates. The question of "basic knowledge" in teaching anatomy during medical training arises as a need for education of future clinical doctors. Nowadays, two extreme views in teaching anatomy can be recognized: one adopted some pure anatomists who feel their existence threatened even by the idea of any reduction in their field, and one by some morphologists exclusively interested in cellular biology, who consider that classical anatomy is of no interest, since it has been exhausted as a field for research. An intermediate position is taken by some clinicians, who maintain that anatomy is indispensable but seek a severe reduction in the content to what they consider to be necessary. The above mentioned need for clinicians was reflected in recommendations of Education Committee of the General Medical Council (GMC) which in short, could be characterized by: the substantial reduction of factual information, the increase of student learning and the emphasis of clinically applied anatomy with its integration to the general medical education. GMC delegated the Department of Anatomy at the University of Cambridge by the developing of the new anatomy course. This new course was for the first time introduced in school year 1998-1999. In this study are presented ways and methods of undergraduate anatomy teaching at the University of Cambridge. These educational principles could serve as a model for teaching anatomy during its transformation in other medical faculties. PMID- 10824418 TI - Four applications of the anesthetic cut-off effect. AB - This article presents and analyses several cases in which the use of the cut-off effect is useful. It starts from the fact that an anesthetic effect of homologous agents is always expressible as a function of their chain lengths and that the cut-off point is a point at which the function vanishes. We then investigate four categories of results: (i) whole body effects. (ii) Cases in which the anesthetics affect the Hodgkin-Huxley parameters of a nerve. (iii) Molecular mechanisms of anesthetic action. (iv) The physical chemistry of the anesthetic process. Our discussion shows that it is possible to incorporate these apparently remote results into one framework. It also shows how to compare results that were gathered by independent measuring methods. In some instances we suggest an interpretation, in others we suggest a further gathering of experimental data. One of the deductions indicates that a weakness exists in the lipid theories of anesthesia. PMID- 10824419 TI - On estimating the probability of aperiodic outbursts of microbial populations from their fluctuating counts. AB - The irregular sequence of counts of a microbial population, in the absence of observable corresponding environmental changes (e.g., temperature), can be regarded as reflecting the interplay of several unknown or random factors that favor or inhibit growth. Since these factors tend to balance one another, the fluctuations usually remain within bounds, and only by a coincidence--when all or most act in unison--does an 'outburst' occur. This situation can be represented mathematically as a sequence of independent random variables governed by a probability distribution. The concept was applied to reported microbial counts of ground meat and wastewater. It is found that the lognormal distribution could serve as a model, and that simulations from this model are indistinguishable from actual records. The parameters of the lognormal (or other) distribution can then be used to estimate the probability of a population outburst, i.e., an increase above a given threshold. Direct estimation of the outburst probability based on frequency of occurrence is also possible, but in some situations requires an impractically large number of observations. We compare the efficiency of these two methods of estimation. Such methods enable translation of irregular records of microbial counts into actual probabilities of an outburst of a given magnitude. Thus, if the environment remains 'stable' or in dynamic equilibrium, the fluctuations should not be regarded merely as noise, but as a source of information and an indicator of potential population outbursts even where obvious signs do not exist. PMID- 10824420 TI - Mathematical model for early development of the sea urchin embryo. AB - In Xenopus and Drosophila, the nucleocytoplasmic ratio controls many aspects of cell-cycle remodeling during the transitory period that leads from fast and synchronous cell divisions of early development to the slow, carefully regulated growth and divisions of somatic cells. After the fifth cleavage in sea urchin embryos, there are four populations of differently sized blastomeres, whose interdivision times are inversely related to size. The inverse relation suggests nucleocytoplasmic control of cell division during sea urchin development as well. To investigate this possibility, we developed a mathematical model based on molecular interactions underlying early embryonic cell-cycle control. Introducing the nucleocytoplasmic ratio explicitly into the molecular mechanism, we are able to reproduce many physiological features of sea urchin development. PMID- 10824421 TI - Self-organization, scale and stability in a spatial predator-prey interaction. AB - Simple predator-prey models often predict extreme instability in interactions where the prey are depressed well below their carrying capacity. Although the behaviour of some laboratory systems conforms to this pattern, field and mesocosm studies generally show prolonged co-existence of prey and predator. Prominent among the possible causes of this discrepancy are the effects of spatial heterogeneity. In this paper we show that both discrete and continuous representations of the spatial Rosenzweig-McArthur model with immobile prey can be stabilized by self-organized prey heterogeneity. This concordance of behaviour closely parallels that which we have previously established in the context of invasion waves. We use the continuous model variant to calculate the characteristic spatial scales of the self-organized structures. The discrete variant forms the basis of a simulation study demonstrating the variety of stable structures and elucidating their relation to the history of the system. We note that all stable prey distributions take the form of a network of occupied patches separated by prey-free regions, and liken the process which generates such assemblages to the formation of a landscape mozaic. PMID- 10824423 TI - Fast subsystem bifurcations in strongly coupled heterogeneous collections of excitable cells. AB - A continuum model for a heterogeneous collection of excitable cells electrically coupled through gap junctions is introduced and analysed using spatial averaging, asymptotic and numerical techniques. Heterogeneity is modelled by imposing a spatial dependence on parameters which define the single cell model and a diffusion term is used to model the gap junction coupling. For different parameter values, single cell models can exhibit bursting, beating and a myriad of other complex oscillations. A procedure for finding asymptotic estimates of the thresholds between these (synchronous) behaviors in the cellular aggregates is described for the heterogeneous case where the coupling strength is strong. This procedure is tested on a model of a strongly coupled heterogeneous collection of bursting and beating cells. Since isolated pancreatic beta-cells have been observed to both burst and beat, this test of the spatial averaging techniques provides a possible explanation to measured discrepancies between the electrical activities of isolated beta-cells and coupled collections (islets) of beta-cells. PMID- 10824422 TI - Using parasites as biological tags of fish populations: a dynamical model. AB - A simple model of macro-parasitic infections has been used to evaluate the potential use of parasites as biological tags of fish populations. In the model, the parasite-host interaction is regulated by a birth-death process, and parasites can only be acquired by the non-specific migratory host population in a particular area of the space domain. In this case, we show that parasites can be successfully used for stocks identification and to describe the migratory routes taken by some marine fish species. PMID- 10824424 TI - Dynamics of one-pass germinal center models: implications for affinity maturation. AB - During an immune response, the affinity of antibodies that react with the antigen that triggered the response increases with time, a phenomenon known as affinity maturation. The molecular basis of affinity maturation has been partially elucidated. It involves the somatic mutation of immunoglobulin V-region genes within antigen-stimulated germinal center B cells and the subsequent selection of high affinity variants. This mutation and selection process is extremely efficient and produces large numbers of high affinity variants. Studies of the architecture of germinal centers suggested that B cells divide in the dark zone of the germinal center, then migrate to the light zone, where they undergo selection based on their interaction with antigen-loaded follicular dendritic cells, after which they exit the germinal center through the mantle zone. Kepler and Perelson questioned this architecturally driven view of the germinal center reaction. They, as well as others, argued that the large number of point mutations observed in germinal center B cell V-region genes, frequently 5 to 10 and sometimes higher, would most likely render cells incapable of binding the antigen, if no selection step was interposed between rounds of mutations. To clarify this issue, we address the question of whether a mechanism in which mutants are generated and then selected in one pass, with no post-selection amplification, can account for the observed efficiency of affinity maturation. We analyse a set of one-pass models of the germinal center reaction, with decaying antigen, and mutation occurring at transcription or at replication. We show that under all the scenarios, the proportion of high affinity cells in the output of a germinal center varies logarithmically with their selection probability. For biologically realistic parameters, the efficiency of this process is in clear disagreement with the experimental data. Furthermore, we discuss a set of, possibly counterintuitive, more general features of one-pass selection models that follow from our analysis. We believe that these results may also provide useful intuitions in other cases where a population is subjected to selection mediated by a selective force that decays over time. PMID- 10824425 TI - Equilibrium stability of single-species metapopulations. AB - We investigate the effect of migration between local populations of a single discrete-generation species living in a ring or an array of habitats. The commonly used symmetric dispersal assumption is relaxed to include the biologically more reasonable asymmetric dispersion. It is demonstrated analytically that density independent migration has no effect on the equilibrium stability of individual populations. However, the positive equilibrium may be destabilizing if the migration is density dependent in such a way that it increases with increasing population density at the source patch. PMID- 10824426 TI - Parathyroid hormone temporal effects on bone formation and resorption. AB - Parathyroid hormone (PTH) paradoxically causes net bone loss (resorption) when administered in a continuous fashion, and net bone formation (deposition) when administered intermittently. Currently no pharmacological formulations are available to promote bone formation, as needed for the treatment of osteoporosis. The paradoxical behavior of PTH confuses endocrinologists, thus, a model bone resorption or deposition dependent on the timing of PTH administration would de mystify this behavior and provide the basis for logical drug formulation. We developed a mathematical model that accounts for net bone loss with continuous PTH administration and net bone formation with intermittent PTH administration, based on the differential effects of PTH on the osteoblastic and osteoclastic populations of cells. Bone, being a major reservoir of body calcium, is under the hormonal control of PTH. The overall effect of PTH is to raise plasma levels of calcium, partly through bone resorption. Osteoclasts resorb bone and liberate calcium, but they lack receptors for PTH. The preosteoblastic precursors and preosteoblasts possess receptors for PTH, upon which the hormone induces differentiation from the precursor to preosteoblast and from the preosteoblast to the osteoblast. The osteoblasts generate IL-6; IL-6 stimulates preosteoclasts to differentiate into osteoclasts. We developed a mathematical model for the differentiation of osteoblastic and osteoclastic populations in bone, using a delay time of 1 hour for differentiation of preosteoblastic precursors into preosteoblasts and 2 hours for the differentiation of preosteoblasts into osteoblasts. The ratio of the number of osteoblasts to osteoclasts indicates the net effect of PTH on bone resorption and deposition; the timing of events producing the maximum ratio would induce net bone deposition. When PTH is pulsed with a frequency of every hour, the preosteoblastic population rises and decreases in nearly a symmetric pattern, with 3.9 peaks every 24 hours, and 4.0 peaks every 24 hours when PTH is administered every 6 hours. Thus, the preosteoblast and osteoblast frequency depends more on the nearly constant value of the PTH, rather than on the frequency of the PTH pulsations. Increasing the time delay gradually increases the mean value for the number of osteoblasts. The osteoblastic population oscillates for all intermittent administrations of PTH and even when the PTH infusion is constant. The maximum ratio of osteoblasts to osteoclasts occurs when PTH is administered in pulses of every 6 hours. The delay features in the model bear most of the responsibility for the occurrence of these oscillations, because without the delay and in the presence of constant PTH infusions, no oscillations occur. However, with a delay, under constant PTH infusions, the model generates oscillations. The osteoblast oscillations express limit cycle behavior. Phase plane analysis show simple and complex attractors. Subsequent to a disturbance in the number of osteoblasts, the osteoblasts quickly regain their oscillatory behavior and cycle back to the original attractor, typical of limit cycle behavior. Further, because the model was constructed with dissipative and nonlinear features, one would expect ensuing oscillations to show limit cycle behavior. The results from our model, increased bone deposition with intermittent PTH administration and increased bone resorption with constant PTH administration, conforms with experimental observations and with an accepted explanation for osteoporosis. PMID- 10824427 TI - Population extinction and quasi-stationary behavior in stochastic density dependent structured models. AB - Density-independent and density-dependent, stochastic and deterministic, discrete time, structured models are formulated, analysed and numerically simulated. A special case of the deterministic, density-independent, structured model is the well-known Leslie age-structured model. The stochastic, density-independent model is a multitype branching process. A review of linear, density-independent models is given first, then nonlinear, density-dependent models are discussed. In the linear, density-independent structured models, transitions between states are independent of time and state. Population extinction is determined by the dominant eigenvalue lambda of the transition matrix. If lambda < or = 1, then extinction occurs with probability one in the stochastic and deterministic models. However, if lambda > 1, then the deterministic model has exponential growth, but in the stochastic model there is a positive probability of extinction which depends on the fixed point of the system of probability generating functions. The linear, density-independent, stochastic model is generalized to a nonlinear, density-dependent one. The dependence on state is in terms of a weighted total population size. It is shown for small initial population sizes that the density-dependent, stochastic model can be approximated by the density independent, stochastic model and thus, the extinction behavior exhibited by the linear model occurs in the nonlinear model. In the deterministic models there is a unique stable equilibrium. Given the population does not go extinct, it is shown that the stochastic model has a quasi-stationary distribution with mean close to the stable equilibrium, provided the population size is sufficiently large. For small values of the population size, complete extinction can be observed in the simulations. However, the persistence time increases rapidly with the population size. PMID- 10824428 TI - Calculation of the cumulative distribution function of the time to a small observable tumor. AB - Multistage mathematical models of carcinogenesis (when applied to tumor incidence data) have historically assumed that the growth kinetics of cells in the malignant state are disregarded and the formation of a single malignant cell is equated with the emergence of a detectable tumor. The justification of this simplification is, from a mathematical point of view, to make the estimation of tumor incidence rates tractable. However, analytical forms are not mandatory in the estimation of tumor incidence rates. Portier et al.(1996b, Math. Biosci. 135, 129-146) have demonstrated the utility of the Kolmogorov backward equations in numerically calculating tumor incidence. By extending their results, the cumulative distribution function of the time to a small observable tumor may be numerically obtained. PMID- 10824429 TI - On the relationship between the Hill coefficients for steady-state and transient kinetic data: a criterion for concerted transitions in allosteric proteins. AB - A frequently used measure for the extent of cooperativity in ligand binding by allosteric proteins is the Hill coefficient. Hill coefficients can be measured for steady-state kinetic data and also for transient kinetic data. Here, the relationship between the two types of Hill coefficients is analysed. It is shown that a value of 1 for the ratio of the two Hill coefficients is a test for a concerted ligand-induced transition between two conformations of the protein, in accordance with the Monod-Wyman-Changeux model. A value of 1 for this ratio has recently been observed for a series of chaperonin GroEL mutants suggesting that ATP-induced allosteric transitions in this protein are concerted. PMID- 10824430 TI - Modeling transcriptional control in gene networks--methods, recent results, and future directions. AB - Mathematical models are useful for providing a framework for integrating data and gaining insights into the static and dynamic behavior of complex biological systems such as networks of interacting genes. We review the dynamic behaviors expected from model gene networks incorporating common biochemical motifs, and we compare current methods for modeling genetic networks. A common modeling technique, based on simply modeling genes as ON-OFF switches, is readily implemented and allows rapid numerical simulations. However, this method may predict dynamic solutions that do not correspond to those seen when systems are modeled with a more detailed method using ordinary differential equations. Until now, the majority of gene network modeling studies have focused on determining the types of dynamics that can be generated by common biochemical motifs such as feedback loops or protein oligomerization. For example, these elements can generate multiple stable states for gene product concentrations, state-dependent responses to stimuli, circadian rhythms and other oscillations, and optimal stimulus frequencies for maximal transcription. In the future, as new experimental techniques increase the ease of characterization of genetic networks, qualitative modeling will need to be supplanted by quantitative models for specific systems. PMID- 10824431 TI - Mathematical modelling of juxtacrine patterning. AB - Spatial pattern formation is one of the key issues in developmental biology. Some patterns arising in early development have a very small spatial scale and a natural explanation is that they arise by direct cell-cell signalling in epithelia. This necessitates the use of a spatially discrete model, in contrast to the continuum-based approach of the widely studied Turing and mechanochemical models. In this work, we consider the pattern-forming potential of a model for juxtacrine communication, in which signalling molecules anchored in the cell membrane bind to and activate receptors on the surface of immediately neighbouring cells. The key assumption is that ligand and receptor production are both up-regulated by binding. By linear analysis, we show that conditions for pattern formation are dependent on the feedback functions of the model. We investigate the form of the pattern: specifically, we look at how the range of unstable wavenumbers varies with the parameter regime and find an estimate for the wavenumber associated with the fastest growing mode. A previous juxtacrine model for Delta-Notch signalling studied by Collier et al. (1996, J. Theor. Biol. 183, 429-446) only gives rise to patterning with a length scale of one or two cells, consistent with the fine-grained patterns seen in a number of developmental processes. However, there is evidence of longer range patterns in early development of the fruit fly Drosophila. The analysis we carry out predicts that patterns longer than one or two cell lengths are possible with our positive feedback mechanism, and numerical simulations confirm this. Our work shows that juxtacrine signalling provides a novel and robust mechanism for the generation of spatial patterns. PMID- 10824432 TI - Time-dependent closed form solutions for fully competitive enzyme reactions. AB - An analytic formalism developed earlier to describe the time evolution of the basic enzyme reaction is extended to fully competitive systems. Time-dependent closed form solutions are derived for the three nominal cases of competition: even, slow and fast inhibitors, allowing for the first time the complete characterization of the reactions. In agreement with previous work, the time independent Michaelis-Menten approach is shown to be inaccurate when a fast inhibitor is present. The validity of the quasi-steady-state approximation on which the present framework is based is also revised. PMID- 10824433 TI - Synchronism in a metapopulation model. AB - We consider a spatially explicit meta-population model with interaction among the two nearest neighbors to relate, with a simple mathematical expression, chaos in the local, uncoupled, populations, the degree of interaction among patches, size of the meta-population, and the stability of the synchronized attractor. Since synchronism is strongly correlated with extinction, our results can provide useful information on factors leading to population extinction. PMID- 10824434 TI - Rate estimation for a simple movement model. AB - This paper introduces a simple stochastic model for waterfowl movement. After outlining the properties of the model, we focus on parameter estimation. We compare three standard least squares estimation procedures with maximum likelihood (ML) estimates using Monte Carlo simulations. For our model, little is gained by incorporating information about the covariance structure of the process into least squares estimation. In fact, misspecifying the covariance produces worse estimates than ignoring heteroscedasticity and autocorrelation. We also develop a modified least squares procedure that performs as well as ML. We then apply the five estimators to field data and show that differences in the statistical properties of the estimators can greatly affect our interpretation of the data. We conclude by highlighting the effects of density on per capita movement rates. PMID- 10824435 TI - Quantitative analysis and model simplification of an epidemic model with primary and secondary infection. AB - Models of particular epidemiological systems can rapidly become complicated by biological detail which can obscure their essential features and behaviour. In general, we wish to retain only those components and processes that contribute to the dynamics of the system. In this paper, we apply asymptotic techniques to an SEI-type model with primary and secondary infection in order to reduce it to a much simpler form. This allows the identification of parameter groupings discriminating between regions of contrasting dynamics and leads to simple approximations for the model's transient behaviour. These can be used to follow the evolution of the developing infection process. The techniques examined in this paper will be applicable to a large number of similar models. PMID- 10824436 TI - Benjamin West's portrait of Joseph Banks. PMID- 10824437 TI - Montagu's variolation. AB - Lady Mary Wortley Montagu is sometimes mentioned by both medical and literary historians as the introducer to England of smallpox inoculation. Usually, the story is garbled by confusion with Edward Jenner's later invention, vaccination. Some historians have rejected her claim, arguing that the credit belongs to the medical establishment of the day. So just how much importance has this gifted amateur in the story of medical science? PMID- 10824438 TI - Johann and Elizabeth Hevelius, astronomers of Danzig. AB - Elizabeth Hevelius (1647-1693) was the second wife of Johann Hevelius, the renowned astronomer of Danzig, and assisted with his observations from the first years of her marriage. Hevelius wrote of her in his books as an able collaborator and she is portrayed in one of them observing with him. She brought out his final, posthumous work. With Johann, she received many notable visitors (including Edmond Halley) and observed with some of them at Danzig. She is the first woman astronomer of whom we have any record. PMID- 10824439 TI - Artists as scientists: nature and realism in early modern Europe. AB - In about 1400, northern European artists suddenly began to depict the natural and human world in a 'realistic' or 'naturalistic' manner. At about the same time, new ideas about how to describe nature realistically emerged among scholars investigating the natural world. Over the next two centuries, this new approach to nature (which eventually became known as 'science') and the belief that it could provide a realistic depiction of nature transformed human attitudes to nature and the material world. Artisans or craftspeople were central to this transformation and thus more important than is usually recognized in forming the new attitudes that characterized the Scientific Revolution. PMID- 10824440 TI - Biochemistry comes of age: a century of endeavour. AB - Biochemistry began as an identifiable, separate discipline at the beginning of the twentieth century. This article traces its early development, focusing on three pioneers--Eduard Buchner, Arthur Croft Hill and Arthur Harden. PMID- 10824441 TI - Chemical-warfare techniques for insect control: insect 'pests' in Germany before and after World War I. AB - During World War I, chemical-warfare practices were introduced into economic entomology in Germany. Fritz Haber, 'the father of chemical warfare', realized that Germany could not win the war and thus looked for 'civilian' uses for his chemical arsenal. Before the war, there was a rhetoric of dangerous 'masses' of insects but the large-scale techniques needed to deal with them had not been developed. The gap between rhetoric and practices enabled entomology to integrate chemical weapons into its working methods. This article traces transformations in the ways of seeing insects and their control from the mid-nineteenth century to after World War I. PMID- 10824442 TI - The understanding of monsters at the Royal Society in the first half of the eighteenth century. AB - In tune with an Enlightenment sensibility that focused on the search for order and regularities, monsters were given a marginal position in eighteenth-century medical works. By contrast, they had an important place at the Royal Society during the second half of the century. This article first focuses on the general interest in monsters within the context of the natural historical agenda and corporate activity of the Society and then addresses the medical understanding of monsters by members of the Society. Finally, it discusses some of the moral and social implications of their medicalization in eighteenth century England. PMID- 10824443 TI - [Speech delivered in the anniversary celebration of the Mexican Association of Gynecology and Obstetrics. February 9, 2000. Commemorative session of the 55th Anniversary. The AMGO and the professional practice of the specialty]. PMID- 10824444 TI - [Sociodemographic factors associated with depression in pregnant adolescents]. AB - The objective of this study is to determine the frequency of depression among pregnant adolescents and explore the potential risk factors associated to the depression. The sample comprised 59 pregnant teenagers recruited from the Program for Pregnant Adolescents in the Hospital General "Dr. Manuel Gea Gonzalez". All subjects completed the Beck Depression Inventory (BDI) and a sociodemographic and medical questionnaire. The frequency of depression was 39%. Sociodemographic correlates of depression were not identified using ratios and 95% confidence limits. Results indicate that the depression is common among pregnant teenagers. Adequate prenatal cares including screening for depression symptoms and development of treatment programs for affected pregnant adolescents are of particular importance for young mothers. In further studies is necessary to explore the impact of family environment. PMID- 10824445 TI - [Relationship between pregestational body mass index and body composition in the immediate puerperium]. AB - To determine the correlation between pregestational body mass index and certain body composition parameters measured by bioelectric impedance (percentage of fat weight, fat mass, and lean weight) and to compare the body composition of women in immediate puerperal period with non-pregnant women, a cross sectional study was done including 86 women in physiologic puerperal period and 90 healthy non pregnant women, from the Instituto Mexicano del Seguro Social, in Chihuahua, Mexico, aged 16 to 40 years old. The variables considered were the body mass index (BMI) before pregnancy, percentage of fat, fat mass, lean weight, total water and bio-resistance. Body composition was measured by bioelectric impedance (BIO) from 4 to 12 hours after delivery. Correlation coefFicient for BMI and percentage of fat was r = 0.66 (p < 0.01), and for BMI and fat mass, it was r = 0.74 (p < 0.01). In non-pregnant women the total water was 35.5 +/- 5 L and in women in the immediate puerperal period 38.5 +/- 5 L (p < 0.01). In conclusion, BMI before pregnancy is a suitable predictor for body composition in the puerperal period. However, it does not discriminate important variables such as total body water, so it may be convenient to use BIA for surveillance of body composition during pregnancy. PMID- 10824446 TI - [Effectiveness and safety of ciclopirox olamine 1% vaginal cream versus terconazole 0.8% vaginal cream in the treatment of genital candidiasis]. AB - A multicenter randomized study was performed. One hundred and seventy patients were selected. The patients were 18 years and older. They presented signs and symptoms of genital candidiasis and had positive smear culture for Candida. Eighty five patients were assigned to receive Cicloprox olamine 1%, and eighty five patients were assigned to receive Terconazol 0.8%. The treatment lasted six days with the objective to compare the clinical and antifungal efficiency and safety with both treatments. The result of mixed efficiency (clinical and microbiology) for Cicloprox olamine was 48 cases (62.3%) at the end of the treatment were cured--day 7-; and at the continuation--day 21-42 of them had the same result (55.3%); improvement was seen at the end in 25 cases (32.5%) and at the continuation 21 cases had the same result (27.6%). For Terconazol the result was 45 cases (61.6%) were cured at the end of the treatment, and at the continuation 39 had the same result (57.4%); improvement was seen at the end in 23 cases (31.5%) and at the continuation 22 cases had the same result (32.4%). We conclude that both treatments are effective and well tolerated for genital candidiasis treatment. PMID- 10824447 TI - [Effect of oral contraceptive administration on the frequency of estrogen receptor and cervical intraepithelial neoplasia]. AB - A study of 56 sexual active women with cervical intraepithelial neoplasia (CIN) was conducted in Mexico City, to determine wether the risk for this condition is altered by using oral contraceptives. Responses of a standardized questionnaire compared 29 patients under oral contraceptive (OC) treatment and 27 control subjects selected from the same screening program, though which the cases were detected. Estrogen receptor (ER) levels were determined in each hyperplastic tissue. The ER frequency was approximately the same in women under oral contraceptive (OC) treatment and in women without it. (P > 0.05). Among other risk factors, first intercourse at an early age, first pregnancy age, number of pregnancies and multiple sexual partners were consistently no significant (p > 0.05) when both groups were compared. In this study, we have found no evidence of an etiologic relationship between neither oral contraceptive usage or non usage and CIN. PMID- 10824448 TI - [Chemotherapy in germinal and epithelial ovarian cancer]. AB - Generically, ovarian cancer represents a group of tumors with diverse biological and clinical behavior. Thus, germinal cells ovarian tumors, in the vast majority of patients are successfully treated utilizing traditional based on cisplatin chemotherapy. Epithelial ovarian cancer, that accounts for 90% of these cases, although sensitive to chemotherapy has not shown satisfactory results. In the search better tumoral response in the treatment of epithelial ovarian cancer new drugs have surged that promise good results, including docetaxel, topotecan and gemcitabine, both as single agents, or in combination with other therapies utilizing monoclonal antibodies. PMID- 10824449 TI - [Vaginal evisceration. Report of a case and review of the literature]. AB - A case report of small bowel penetration through the cult the sac in a 23 year old female patient is presented and the management is discussed, with review of the world literature. PMID- 10824450 TI - [Assessment of effectiveness of and tolerance to oxybutynin in the treatment of unstable bladder in women]. AB - Unstable bladder is a frequent syndrome in women and is due in the most part because of detrusor involuntary contractions, mainly due to detrusor denervation, which produces voiding hypersensitivity and loss of cortical inhibition control, clinical manifestations are: frequency, nicturia, urgency and urge incontinence. Historically the most effective treatment has been muscular relaxing agents and anticholinergic agents. We present a prospective, double blind, cross, placebo control study to evaluate efficacy and tolerance of oxybutynin in women with unstable bladder. We included 44 adult women with unstable bladder, 22 unitially received oxybutynin 5 mg t.i.d. and 22 placebo 5 mg t.i.d. aleatory in both groups through 6 weeks, later wash-out period was performed and those women in which initially received oxybutynin were administered placebo and those women in which initially received placebo, were administered oxybutynin, for another six weeks. Five patients which initiated the study with oxybutynin abandoned the study, 2 of them for intolerance and 3 for unknown causes. Two women in which initially received placebo abandoned follow-up too. A total of 74 subjects (37 for each branch of study) had an age average of 51.7. Symptoms scoring decreased from 13 to 11 and 6 points with placebo and oxybutynin respectively (p = 0.001). The analog visual scale of symptoms decreased from 77% to 62.5% and 40% with placebo and oxybutynin respectively (p = 0.003). The overall rate of improvement evaluated through symptoms scoring was from 27% with placebo and 72.9% with oxybutynin (p = 0.000) and evaluated through analog visual scale of symptoms was from 40% with placebo and 78.3% with oxybutynin (p = 0.002). The vesical volume at first voiding sensation increased from 129 ml to 134 ml and 187 ml with placebo and oxybutynin respectively (p = 0.021) and the maximum cystometric capacity increased from 231 ml to 236 ml and 301 ml with placebo and oxybutynin respectively. The most frequent adverse effect in both groups was mouth dryness and it presented in 7 (19%) and 34 (91%) patients with placebo and oxybutynin respectively (p = 0.000). Only 5 of 44 patients (11.3%) with oxybutynin and 2 of 44 patients (4.4%) with placebo abandoned follow up (p = 0.14). We concluded that oxybutynin improve significantly the unstable bladder symptoms in women, possibly by increasing functional bladder capacity and decreasing voiding sensitivity, with good tolerance of mouth dryness in the majority of patients. PMID- 10824451 TI - Laparoscopic cholecystectomy: experience with 150 consecutive patients in Kurashiki. AB - One hundred and fifty consecutive patients who underwent laparoscopic cholecystectomy (LC) in Kurashiki between March 1991 and December 1997 were studied in order to examine intraoperative procedures, and complications, especially with regard to bile duct injuries and acute cholecystitis, furthermore postoperative management. Nine out of the 150 cases were converted to open cholecystectomy, so that the overall success rate was 94%. One hundred and forty six patients (97%) presented on an elective basis, and the remaining four patients were admitted as acute cholecystitis. One hundred and forty-four patients had gallbladder stones. All of seven patients had adenomyomatosis with stones (4 cases) and without stones (3 cases). Nine patients had benign polyps of the gallbladder with stones (6 cases) and without stones (3 cases). Seven patients with choledocholithiasis were treated successfully with endoscopic sphincterotomy preoperatively. In six out of eight patients with acute cholecystitis, among whom 4 were treated preoperatively with percutaneous transhepatic gallbladder aspiration (PTGBA), LC was conducted. Major complications included 3 cases of bile duct injuries (2%). Twenty cases (14) of bleeding encountered during operation were controlled easily under laparoscopy. Postoperative oral feeding was started in nearly all cases on the 1st postoperative day. In conclusion, LC seems to offer significant advantages to patient recovery but attention to bile duct injury and the judgement for the timing of conversion to open cholecystectomy is considered necessary for a successful LC. PMID- 10824452 TI - Newly designed eccentric roller type total artificial heart. AB - We have produced an eccentric roller type total artificial heart (ERTAH). As the first step in the development of this ERTAH, we conducted simulations such as a numerical simulation, a mock test, and an acute animal experiment using DeBakey roller pumps to analyze the left-right balance during its operation. The next step was redesigning the blood chambers to improve energy efficiency and implanting the ERTAH with an interatrial shunt into an animal for evaluation of the in vivo performance of the device. In the simulations, shunt flow through the bronchial arteries was approximately 500-600 ml/min, and the interatrial resistance was varied from 2.9-7.7 mmHg.min/liter. Redesigning the blood chambers resulted in the mock test in a 20% increase in energy efficiency, about a two fold increase in cardiac output and improved durability compared to the previous type in the mock test. In the animal experiment the ERTAH operated with a left flow rate of 6.0 liters/min and a right flow rate of 5.4 liters/min. The interatrial shunt flow rate was 250-400 ml/min. Creating an interatrial shunt is a useful method for balancing the blood flow between the left and right heart of the ERTAH. A decrease in friction resistance and the prevention of backward flow resulted in an increase in energy efficiency, cardiac output, and improved durability, in spite of downsizing the blood chambers. PMID- 10824453 TI - Comparison of properties of slow pathway potential between successful and unsuccessful radiofrequency applications in patients who underwent catheter ablation for atrioventricular nodal reentrant tachycardia. AB - Findings concerning selective slow pathway radiofrequency ablation for atrioventricular nodal reentrant tachycardia (AVNRT) using the slow pathway potential (SPP) guided method are reported. The electrogram at the SPP recording site showed double potentials consisting of the atrial potential (A) and SPP. However, SPP is usually recorded widely in the right atrial posteroseptal region. To examine whether there was any characteristic marker in the electrogram at the SPP recording site specific to successful RF application, the properties of SPP and its anatomical locations in both successful (S) (38 sites) and unsuccessful (UN) (28 sites) application sites were analyzed in 38 patients who underwent SPP guided ablation. The distance between the upper margin of the coronary sinus ostium (UCSO) and the ablation catheter (ABL) (DUCSO-ABL) was shorter in S than in UN (2.3 +/- 6.3 mm vs. 9.0 +/- 5.2 mm below the level of UCSO, p < 0.001). The interval between A and SPP (A-SPP) was longer in S than in UN (44.2 +/- 9.9 msec vs. 24.0 +/- 7.0 msec, p < 0.001). RF applications at the more anterior sites with longer A-SPP were more successful than at other sites. The sensitivity and specificity of A-SPP (more than 40 msec) were superior to those of DUCSO-ABL (within 5 mm) as the marker for the successful application (sensitivity; 73.7% v.s. 68.4%, specificity; 100% v.s. 82.1%, respectively). In conclusion, the sites with longer A-SPP might be specific for successful ablation. PMID- 10824454 TI - Gene therapy for murine renal cell carcinoma using genetically engineered tumor cells to secrete interleukin-12. AB - To determine the possibility of gene therapy for renal cell carcinoma (RCC) using interleukin-12 (IL-12), we prepared genetically engineered murine RCC cells (Renca) which secrete IL-12 and evaluated the usefulness of these cells as a tumor vaccine. The IL-12 gene was transduced using MFG retroviral vector. The in vitro characteristics of transfectants--i.e., cell proliferation and expression of surface antigens--were then examined. In vivo tumorigenicity was assessed by subcutaneously injecting each type of cell in syngenic BALB/c mice. For the challenge experiments, the mice rejecting previously injected Renca IL-12 cells were rechallenged with parental cells. To determine the antitumor effect at remote sites, mice were injected with parental cells into the left flank, and then either Renca IL-12 or parental cells were inoculated into the opposite site on day 0 or 1. The transfected cells can secrete 146.7 ng/ml/10(6)cells/48 hr of IL-12, as confirmed here by bioassay. The in vitro characteristics of the transfectants were not altered, but in vivo tumorigenicity was significantly reduced. Of the 21 mice that rejected Renca IL-12 cells, 9 failed to develop tumors after the challenge with parental cells. In the mice treated with Renca IL 12 as a vaccine, both number and tumor volume of the mice that developed tumors at remote sites were reduced. IL-12 secreting Renca cells conferred both protective immunity to parental cells and delay of tumor growth at remote sites, indicating that IL-12 secreting Renca cells are a feasible candidate for use in gene therapy of RCC. PMID- 10824455 TI - Visual event-related potentials in progressive supranuclear palsy. AB - The purpose of this study is to assess the cognitive function in early stages of progressive supranuclear palsy (PSP). Visual event-related potentials (visual ERPs) were examined in five non-demented patients with PSP and seven control subjects. ERPs were recorded using Figure (non-verbal) and Kanji (verbal) oddball paradigms. The latencies and amplitudes of N100 and P300 components were not significantly different between the PSP and control groups. However, the score of Hasegawa's dementia scale-revised (HDSR) was correlated with P300 amplitudes in the Figure task only in the PSP group. Since the P300 amplitude is related to attentional resources, changes in visual ERP induced by non-verbal stimuli might be associated with the attentional impairment even in early stages of PSP. In the Kanji task but not in the Figure task, the reaction time was prolonged in the PSP group, and positively correlated with the P300 latency in both groups. Visual verbal stimuli might be a good tool to evaluate the information processing in the early stages of PSP. Recording of visual ERP using both non-verbal and verbal stimuli could be helpful to assess a mild cognitive dysfunction in PSP. PMID- 10824456 TI - Generation of pro-inflammatory and anti-inflammatory cytokines in the gut in zymosan-induced peritonitis. AB - In major systemic inflammation such as severe peritonitis, various pro inflammatory cytokines, such as TNF-alpha, IL-1 beta and IL-6, play important roles in the development of multiple organ dysfunction syndrome (MODS). The purpose of this study was to investigate the outflow of pro-inflammatory and anti inflammatory cytokines from the efferent mesenteric lymphatic vessels under peritonitis. Mesenteric lymph samples were collected from adult male rats at 2, 4, 6, 8 and 10 hr after an intraperitoneal injection of zymosan at a dosage of 0.1 mg/g (non-lethal dose) or 0.5 mg/g (lethal dose). Blood samples were obtained at 10 hr after zymosan administration. The amounts of drained TNF-alpha and IL-6 in the lymph peaked at 2-4 hr and 4-8 hr after zymosan administration, respectively. The amounts of drained IL-10 in the lymph gradually increased until 10 hr. The amounts of drained TNF-alpha and IL-10 in the mesenteric lymph were significantly correlated with the dosage of zymosan. In conclusion, under intraperitoneal inflammation, pro-inflammatory cytokines (TNF-alpha and IL-6) increased in the mesenteric lymph and were drained into circulation. IL-10, one of the anti-inflammatory cytokines, also increased in the mesenteric lymph after several hours' delay and its increase was remarkable in several inflammations. These findings suggested that the gut might be one of the pro-inflammatory and anti-inflammatory cytokine-generating organs under peritonitis. The lymph-drained amounts of each cytokine under peritonitis are considered to differ with the time or severity of inflammation, which may cause different conditions in patients due to the imbalance of pro-inflammatory and anti-inflammatory cytokines. PMID- 10824457 TI - Modulation of ciliary activity by tumor necrosis factor-alpha in cultured sinus epithelial cells. Possible roles of nitric oxide. AB - The primary function of well-differentiated ciliated epithelium in the paranasal sinus is to eliminate harmful agents through the beating action of cilia. Respiratory epithelium also contributes to local inflammatory processes through the release of various proinflammatory cytokines. Recently, considerable attention has been focused on the intimate relationship between the cytokine dependent regulation of the ciliary beat frequency (CBF) and intra-cellular production of nitric oxide (NO) in ciliated epithelial cells. The aims of this study are to examine the effect of tumor necrosis factor-alpha (TNF-alpha), one of the major proinflammatory cytokines, on the ciliary activity of human sinus epithelial cells and to assess the hypothesis that NO is involved in this regulatory mechanism. Human maxillary or ethmoidal sinus mucosa (n = 23) were cultured by the explant-outgrowth method. CBF of the outgrowth ciliated cells was measured by the photoelectrical method before and after being treated with TNF alpha (0.1, 1 and 10 ng/ml) or dexamethasone (10(-6) M and 10(-7) M). We also investigated the expression of nitric oxide synthase (NOS) isoforms, enzymes responsible for NO synthesis, by fluorescent immunohistochemistry. TNF-alpha increased CBF at relatively low concentrations (0.1 and 1 ng/ml) and decreased CBF at a high concentration (10 ng/ml). Dexamethasone decreased CBF at a concentration of 10(-6) M. Fluorescent immunohistochemistry demonstrated that the expression of inducible NOS was augmented by TNF-alpha and attenuated by dexamethasone, whereas that of endothelial NOS remained unchanged. We conclude that human sinus epithelial cells potentially contribute to the inflammatory process by regulating their ciliary motility through an NO-dependent pathway. Proinflammatory cytokines and steroids are able to modulate this mechanism by the induction or inhibition of expression of different NOS isoforms. PMID- 10824458 TI - Albumin permeability across endothelial cell monolayer exposed to reactive oxygen intermediates: involvement of reversible functional alteration of the cell membrane Ca2+ channels. AB - This study was designed to test the idea that the redox state of sulfhydryl (SH) groups in cell-membrane Ca2+ channels plays a pivotal role in Ca2+ influx, which in turn causes an increase in albumin permeability across the cultured monolayer of porcine pulmonary artery endothelial (PPAE) cells exposed to xanthine/xanthine oxidase (X/XO). Albumin permeability as well as the concentration of intracellular Ca2+ ([Ca2+]i) was increased by X/XO. A H2O2 scavenger (catalase), an iron chelator (o-phenanthroline), and a hydroxyl radical scavenger (dimethyl sulfoxide) inhibited these changes provoked by X/XO, in which intracellular iron catalyzed hydroxyl radical generation was suggested to be involved. The increase in albumin permeability and [Ca2+]i continued once the PPAE cells were exposed to X/XO. The [Ca2+]i was decreased by a Ca2+ channel blocker, Ni2+, while the removal of Ni2+ increased [Ca2+]i again, suggesting the sustained Ca2+ influx through cell-membrane Ca2+ channels was responsible for the [Ca2+]i elevation. Ni2+ failed to inhibit albumin permeability sustained after the removal of X/XO. In contrast, SH-reducing agents (dithiothreitol and glutathione) inhibited the sustained permeability as well as Ca2+ influx. We concluded that the redox alteration of SH-groups in cell-membrane Ca2+ channels was involved in the increase in albumin permeability after exposure of the endothelial cells to oxidative stress. PMID- 10824459 TI - Effects of N-(4-hydroxyphenyl) retinamide on urokinase-type plasminogen activator and plasminogen activator inhibitor-1 in prostate adenocarcinoma cell lines. AB - Previous investigations have demonstrated that a synthetic retinoid, N-(4 hydroxyphenyl) retinamide (4-HPR), inhibits the invasion of prostate adenocarcinoma in vitro. Urokinase-type plasminogen activator (uPA) is a prerequisite for tumor invasion. The purpose of this study was to evaluate the effects of 4-HPR on uPA and plasminogen activator inhibitor-1 (PAI-1) in prostate cancer. Human prostate adenocarcinoma cell lines, TSU-PR1 and PC3, were grown in serum-free media containing 4-HPR. Cellular mRNA and protein were subsequently extracted. Northern blot analysis, enzyme-linked immunosorbent assay (ELISA) and chromogenic functional analysis were performed on the samples. Administration of 10(-6) M 4-HPR for 3 days resulted in an increase in uPA mRNA expression (TSU PR1: 391%, PC3: 356%), and a simultaneous increase in PAI-1 mRNA expression (TSU PR1: 217%, PC3: 235%) was observed. ELISA concomitantly demonstrated a significant increase (p < 0.05) in uPA protein in the conditioned media (TSU-PR1: 134%, PC3: 139%) and cell lysates (TSU-PR1: 284%, PC3: 255%). Both cell lines demonstrated a significant increase (p < 0.05) in PAI-1 protein in the conditioned media (TSU-PR1: 152%, PC3: 167%) and cell lysates (TSU-PR1: 170%, PC3: 222%). Concentrations below 10(-6) M failed to alter the protein production of either uPA or PAI-1. The functional uPA assay demonstrated a reduction of the proteolytic activity of uPA (TSU-PR1: 13%, PC3: 7%) in cell lysates of 10(-6) M 4 HPR (p < 0.05), while there was minimal uPA activity in the conditioned media. 4 HPR stimulates a paradoxical increase in uPA and PAI-1, but the anti-invasive effects of 4-HPR are consistent with the increase in both uPA and PAI-1, resulting in an overall reduction of functional uPA activities. PMID- 10824460 TI - Anti-tumor effect of murine renal cell carcinoma cells genetically modified to express B7-1 combined with cytokine secreting fibroblasts. AB - Recently, many experiments have shown that the expression of the costimulatory molecule B7-1 on tumor cells can induce tumor-specific immunity. These results suggest that tumor cells modified to express costimulatory molecules can be used as a potential tumor vaccine. For this purpose, we transduced B7-1 gene into renal adenocarcinoma cells of spontaneous origin (Renca) in BALB/c mouse using the retroviral vector system. Our results indicated that approximately 60% of cells expressed B7-1 gene product using the retroviral vector system, and that B7 1 transduction did not affect the expression of MHC molecules on tumor cells nor the in vitro growth rate of tumor cells, but only in vivo tumorigenicity. As for the antitumor effect on the remote site, there were no significant differences among parental Renca, Renca lac Z and Renca B7-1 sublines, although tumors grew a little more slowly in the mice injected with Renca B7-1 cells as a vaccine. Even if the growth of tumors was significantly delayed in the mice treated by Renca B7 1 as a vaccine combined with the injection of BALB/c3T3 IL-12 near to the tumor on the same or following day, no significant antitumor effects were observed when the Renca B7-1 cells were injected as a vaccine compared with cytokines near the vaccine site. These results indicated that B7-1 gene transduction can decrease the tumorigenicity of murine renal cell carcinoma cells, but fails to induce sufficient antitumor response when it is used as a tumor vaccine. It is necessary to develop immunogenicity, by such means as irradiation or a combination of appropriate cytokines, to stimulate effective tumor immunity in a therapeutic setting. PMID- 10824461 TI - Light and electron microscopic study of peripheral nerve damage in patients with lepromatous leprosy (LL) and borderline lepromatous leprosy (BL). AB - Cutaneous branches of radial nerves in patients with lepromatous leprosy (LL) and borderline lepromatous (BL) were studied by light and electron microscopy. Foamy macrophages were found more or less in the nerve fibers of all leprosy patients and distributed in the epineurial, perineurial and endoneurial areas. In the endoneurium, the foamy macrophages were mainly located in the subperineurial and perivascular spaces. Vacuolated Schwann cells were also found in the nerve fasciculus. In electron microscopy, these foamy macrophages and vacuolated Schwann cells contained numerous small dense materials, irregular in size and shape, considered to be degenerated and fragmented mycobacterium leprae. These dense materials were found also in the cytoplasm of vascular endothelial cells. These findings suggest that mycobacteria enter into the endoneurium via the blood vessels. In our present study, on the other hand, it was very difficult to find the intact mycobacteria in the cytoplasm of the foamy macrophages, Schwann cells or endothelial cells, as well as in the Ziehl-Neelsen staining of paraffin sections. The disappearance of intact bacilli in our present study might have been caused by multi drug therapy. The myelinated nerve fibers were degenerated and disappeared in variable degrees. Degenerative changes of the myelin sheath developed from the outer layer to the inner layer with disarrangement of the lamellar structure. These findings were different from myelin destruction of peripheral nerves in Wallerian degeneration. The degenerative changes of the myelin sheath are caused by degeneration and destruction of Schwann cells in leprosy patients. Fibrosis surrounding myelinated and unmyelinated nerve fibers, i.e., periaxonal fibrosis, was found to a greater or lesser extent in the endoneurium. In the present study, it is still unclear whether the periaxonal fibrosis was due to necrosis of the Schwann cells by infection of mycobacteria or to an autoimmune mechanism such as antiperipheral nerve antibody. However, lamellated concentric fibrosis surrounding regenerative myelinated and unmyelinated nerve fibers with the disappearance of mycobacteria suggests that degenerations and regenerations of nerve axons were repeated during clinical cause. These findings indicated that autoimmune mechanisms play an important role in the pathogenesis of periaxonal fibrosis. PMID- 10824462 TI - Two cases of hypothyroidism complicated by renal dysfunction. AB - We report two patients in whom hypothyroidism was considered to cause renal dysfunction. Case 1 was a 65-year-old woman who stopped taking levothyroxine sodium for hypothyroidism. After 6 months, she developed proteinuria, edema, weight gain, and renal dysfunction. Renal biopsy revealed focal segmental proliferative glomerulonephritis. After re-administration of levothyroxine sodium, thyroid function and renal function both recovered. Case 2 was a 51-year old man who presented with edema, difficulty in swallowing, muscular weakness, and fatigue. We diagnosed hypothyroidism, and focal segmental proliferative glomerulonephritis was revealed by renal biopsy. After administration of levothyroxine sodium, his symptoms resolved and his thyroid function and renal function both improved. Our experience suggests that hypothyroidism should be taken into consideration as one of the causes of renal dysfunction. PMID- 10824463 TI - A case of eosinophilic pleural effusion induced by pancreatothoracic fistula. AB - A 49-year-old man was admitted for evaluation of a left pleural effusion. Thoracenthesis yielded a hemorrhagic pleural effusion with a high percentage of eosinophils (15.9%). Although there were no significant abdominal signs, serological examinations demonstrated a marked increase of pancreatic enzyme activity. Moreover, abdominal CT demonstrated cystic changes between the tail of the pancreas and the spleen. Accordingly ERP was performed under pressure, and contrast medium draining from the pancreas was observed. Pancreatic pleural effusion in this patient consisted of pancreatic juice retained in the thoracic cavity, which resulted from intrapancreatic fistulation connecting to the thoracic cavity due to a pancreatic cyst caused by chronic pancreatitis. The present report indicates that we should investigate the retention of eosinophilic pleural effusion considering not only the possibility of thoracic disease, but also the possibility of a pleural effusion derived from abdominal diseases. PMID- 10824464 TI - Prevalence of HCV & HBV infection amongst HIV seropositive intravenous drug users & their non-injecting wives in Manipur, India. AB - Hepatitis C virus (HCV) and hepatitis B virus (HBV) infections were evaluated among 77 Manipuri couples of whom all husbands were both intravenous drug users (IDUs) and human immunodeficiency virus (HIV) positive. This study showed for the first time a high prevalence of HCV (92%) and HBV (100%) infection amongst the IDUs in Manipur. Stringent control measures to prevent the transmission of hepatitis viruses (B and C) are urgently required in Manipur. PMID- 10824465 TI - Occurrence of sef & pef genes among different serovars of Salmonella. AB - A total of 29 strains of Salmonella enterica belonging to seven serovars isolated from human, animals and birds were used to study the occurrence of Salmonella fimbriae genes (sef and pef) by PCR amplification technique using their specific primers. All the strains (15) of S. Enteritidis were found to carry both sef and pef genes irrespective of the source of isolation. S. Typhimurium strains were found to harbour only pef genes, while S. Gallinarum strains harboured only sef genes. Other serovars namely, S. Newport, S. Kentucky, S. Weltevreden and S. Indiana were negative for both pef and sef genes. The importance of fimbriae in the pathogenesis of salmonellosis is suggested. PMID- 10824466 TI - Kinetics of humoral & cellular immune responses in experimental cysticercosis in pigs infected with Taenia solium. AB - Studies were undertaken to assess the kinetics of antibody responses, lymphocyte transformation to Taenia solium larval antigens (crude soluble extract antigen and antigen B), and T cell subpopulation in piglets following experimental infection. Cysticercosis was established in 1-2 month old piglets after feeding 5,00,000 T. solium eggs per pig. The anti-CD4 and anti-CD8 monoclonal antibodies against swine T cells were raised indigenously. It was observed that at 60 days post infection (PI) there was a significant increase (P < 0.01) in CD4+ T cells without any change in CD8+ T cells. Increased 3H-thymidine uptake was found in infected piglets at 45 days PI using both CSE and antigen B. Kinetics of antibody responses indicated significant increase (P < 0.01) at 15 days PI (with CSE antigen) and 30 days PI (with antigen B) by ELISA. This increase persisted till 90 days PI (the time up to which the animals were followed). It was also observed that the cellular mechanisms were triggered in late stage (60 days PI) as compared to humoral responses (15-30 days PI) and may persist longer as seen by both lymphocyte transformation and T cell subpopulation studies. The study suggests that in cysticercosis, both humoral and cellular mechanisms may play a role in the host defences. PMID- 10824467 TI - Genomic diversity of group A rotavirus RNA from children with acute diarrhoea in Chennai, south India. AB - Group A rotavirus was identified in 51 of 245 (20.8%) cases with acute diarrhoea in Chennai analysed between December 1997 and March 1999. Forty eight of the 51 specimens were subgrouped and serotyped. A total of 110 rotavirus positive specimens (inclusive of 62 rotavirus positive cases reported earlier) were analysed for their subgroup (SG) specificity and genomic profiles. SGI and SGII specificity were detected in 60 per cent and 20 per cent of the cases studied. Twenty two cases showed dual SG specificity (SGI + II). Nine electropherotypic patterns (7 'short' and 2 'long') were observed with a predominance of short pattern in 87 of the 110 (79.1%) positive cases studied. Long electropherotypes were found in 23 (20.9%). Serotyping of the 48 rotavirus positives revealed a higher proportion of serotype-2 (68.8%) followed by serotype-1 (14.6%) and serotype-3 in 1 case. Mixed infection of G1-G2 was observed among 7 cases analysed, which revealed G[2,1], P[4,8] genotype specificity. Dual infection of P[4]-P[8] genotypes was observed in 12 cases with G[2] specificity. PMID- 10824468 TI - Epsilon aminocaproic acid in paediatric cardiac surgery to reduce postoperative blood loss. AB - We have studied the efficacy of epsilon aminocaproic acid in reducing postoperative blood loss in infants and children with congenital cyanotic cardiac anomalies undergoing corrective operative procedures. This prospective study was carried out on 170 infants and children randomly divided into two equal groups. Group A acted as the control group and received normal saline as placebo while group B patients received epsilon aminocaproic acid (100 mg/kg body wt) intravenously slowly soon after anaesthetic induction followed by 100 mg/kg in the cardiopulmonary bypass pump at the time of starting of cardiopulmonary bypass and 100 mg/kg after weaning from bypass over a period of 3 h. In group A the time for sternal closure after separation from bypass and administration of protamine was 75.18 +/- 5.5 min and in group B 50.7 +/- 5.2, (P < 0.001). Blood loss at 24 h in group A was 42.6 +/- 6.9 ml/kg/24 h and in group B 23.7 +/- 5.8 ml/kg/24 h, (P < 0.001). The need for packed red cells in group A was 21.8 +/- 7.1 ml/kg/24 h and in group B 10.7 +/- 7.8 ml/kg/24 h, (P < 0.001). The need for platelet concentrate in group A was 22.0 +/- 6.7 ml/kg/24 h and group B 6.2 +/- 3.2 ml/kg/24 h, (P < 0.001). Fibrin degradation products (split) in group A was 8.2 +/- 0.8 micrograms/ml, and group B 3.8 +/- 1.3 micrograms/ml, (P < 0.001). Reexploration rate was also considerably reduced in group B, 5 of 85 (6%) compared to group A, 13 of 85 (15%), (P < 0.001). It was found that epsilon aminocaproic acid is effective in reducing postoperative blood loss, packed red cells and plasma product requirements in paediatric patients undergoing corrective surgical procedures for congenital cyanotic heart diseases. PMID- 10824469 TI - Effect of renal ischaemia reperfusion on calcium oxalate retention. AB - The effect of ischaemia reperfusion induced renal injury for calcium oxalate deposition under normal and simulated conditions was studied. Male Wistar rats of both control (group I) and urolithic (group II) groups underwent (1 h) unilateral renal artery occlusion and were subjected to 1, 3, 6, 12, 24 and 72 h reperfusion. The group I rats subjected to 1 h renal ischaemia followed by 3 and 6 h reperfusion had significant oxalate retention than that of sham operated controls. In group II, under hyperoxaluric condition, in addition to accumulation of oxalate, calcium oxalate deposits were also observed. The increased retention of calcium oxalate was attributed to increased oxalate binding protein activity, oxalate synthesizing enzymes lactate dehydrogenase and xanthine oxidase activities and accumulation of calcium. Our findings suggested that renal cellular injury produced by ischaemia reperfusion could accelerate calcium oxalate precipitation reaction. PMID- 10824470 TI - Surgical repair of postinfarction ventricular septal rupture. AB - OBJECTIVES: Postinfarction ventricular septal rupture is fatal without surgical repair because of heart failure and secondary multiple organ failure. We investigated surgical results of postinfarction ventricular septal rupture and discussed the surgical strategy of postinfarction ventricular septal rupture. METHODS: Twelve patients (mean age 71.3 +/- 7.4 years, with range from 61 to 81 years) underwent surgical repair of postinfarction ventricular septal rupture, from 1990 to 1998 in our Institute. There were 6 women and 6 men. The ventricular septal rupture was anterior in 10 patients and inferior in 2. The operative technique for anterior ventricular septal rupture was reconstruction of the septum with a Dacron patch after infarctectomy, according to the method of Daggett et al. For posterior ventricular septal rupture, reconstruction of the septum with a Dacron patch after infarctectomy was performed and the ventricular incision was closed with a two-layer patch. Coronary artery bypass grafting was performed in 5 patients for severe proximal coronary artery stenosis using saphenous vein grafts. RESULTS: Overall hospital mortality was 0%. A postoperative residual shunt was recognized in 3 patients, but all were well controlled conservatively and re-operation was not needed. The patients have been followed up for a mean of 59.5 months. There have been two late deaths due to non cardiac problems. Acturial survival rate for the 12 patients was 90% at 1 year and 75% at 5 years. CONCLUSIONS: The Daggett method is simple and fast, and is an effective and reliable technique for the repair of ventricular septal rupture. PMID- 10824471 TI - Results from coronary artery bypass surgery combined abdominal aortic aneurysm repair. AB - OBJECTIVE: Complication from coronary artery disease is a major cause of mortality and morbidity in patients undergoing abdominal aortic aneurysm repair. We report our results from coronary artery bypass surgery performed in combination with abdominal aortic aneurysm repair in patients with coronary artery disease and abdominal aortic aneurysm, each being an indication for an emergency operation. METHODS: Seventeen patients underwent combined coronary artery bypass surgery and abdominal aortic aneurysm repair. The mean age of the patients was 67.6 +/- 5.2 years. Four had left main disease, 8 patients had triple-vessel disease, and 12 had a prior myocardial infarction. The average left ventricular ejection fraction was 0.49 +/- 0.13. The average abdominal aortic aneurysm diameter was 6.2 +/- 1.0 cm (range 4.5-8.0 cm). Thirteen patients underwent coronary artery bypass surgery followed by abdominal aortic aneurysm repair after discontinuation of cardiopulmonary bypass. In the remaining four patients, including one patient with severe left ventricular dysfunction, cardiopulmonary bypass was continued as a circulatory assist until the abdominal aortic aneurysm repair was completed. The left internal thoracic artery was used in 14 patients, and the right internal thoracic artery in one patient. RESULTS: Postoperative surgical complications occurred in three patients (bleeding in one patient requiring reoperation, abdominal subcutaneous wound infection in another and transient neural disorder in the others). There were no surgical or in hospital death. There was no late cardiac complication and no late cardiac death after a mean of 29 months follow-up. CONCLUSIONS: We concluded that combined surgery was reasonable for selected patients with combined coronary artery disease and abdominal aortic aneurysm, each of which is an indication for an urgent operation. The aortic aneurysm repair during cardiopulmonary bypass for patients with severe left ventricular dysfunction was safe and effective. PMID- 10824472 TI - Pedicled pericardial flap for pulmonary artery in adult dogs. AB - OBJECTIVE: Stenosis of extracardiac conduit after reconstruction of right ventricle outflow tract is a serious problem, and the purpose of this study was to identify the suitability of broadly pedicled autologous pericardial flap for the reconstruction of pulmonary artery trunk in adult dogs. METHODS: Eight mongrel dogs had replacement of a pulmonary artery trunk with extracardiac conduit, in which prosthetic vascular graft formed the posterior wall and pedicled (group P, n = 5) or free (group F, n = 3) autologous pericardium created the anterior wall. Six months after the operation, pressure gradient across the conduit were measured and put to death for pathological examinations. RESULTS: Pressure gradient across the conduit in groups F and P was 16.0 +/- 16.8 mmHg and 1.4 +/- 1.7 mmHg respectively. In the microscopic examination, flaps of group P had neo-intimal cells in the innermost layer, abundant cellular component with elastic fiber was seen within the middle layer, and collagen tissue within the outer layer. Conversely, the presence of calcification was shown within collagen fiber layer of all patches in group F. Sparse cellular component and the absence of neo-intimal cells were also observed in group F. CONCLUSIONS: Broadly pedicled pericardial flap is more conceivable to prevent the development of conduit stenosis after right ventricle outflow tract reconstruction, if compared with free pericardial patches. PMID- 10824473 TI - Duplex scanning to assess radial artery suitability for coronary artery bypass grafting. AB - OBJECTIVE: Radial artery suitability in coronary artery bypass grafting was assessed using duplex ultrasonography. SUBJECTS AND METHODS: The vascular condition along the entire radial artery was scanned in 55 patients, measuring the internal diameter and mean flow velocity at the wrist (distally), after ulnar artery branching (proximally), and midway between these 2 points (medially). Distally along the radial and ulnar arteries, the mean flow velocity was determined before and after radial artery occlusion. RESULTS: Atherosclerotic changes were detected in 4 patients. The internal diameter was 3.1 +/- 0.4 mm proximally, 2.7 +/- 0.3 mm medially, 2.4 +/- 0.4 mm distally. The distal flow velocity was 0, and a reverse flow (peak velocity: 11.3 +/- 6.0 cm/s) was observed after the occlusion test in patients with an intact palmar arch, their mean flow velocity, 21.1 +/- 8.9 cm/s, and flow distally along the ulnar artery 58.0 +/- 23.4 ml/min, were higher after the occlusion test than before it 14.7 +/ 6.7 cm/s mean flow and 38.1 +/- 15.9 ml/min distally. This was not observed in patients with an interrupted palmar arch. In 15 patients, radial arteries could not be used because of their small internal diameter, lack of a radial artery, poor vascular condition, or an interrupted palmar arch evaluated using duplex scanning. CONCLUSION: Reliable noninvasive assessment of radial artery anatomy and palmar arch continuity is thus possible using duplex ultrasonography. PMID- 10824474 TI - Early experience of aortic valve replacement with the Freestyle stentless aortic bioprosthesis in elderly patients. AB - OBJECTIVES: Stentless bioprostheses have been gaining popularity in recent years as hemodynamically superior alternatives to conventional stented bioprostheses. METHODS: Between July 1996 and November 1998, 13 patients with aortic valve disease, 7 males and 6 females with a mean age (+/- SD) of 68 +/- 5 years, underwent an aortic valve replacement using the Medtronic Freestyle aortic bioprosthesis. The predominant lesions were stenosis in 8 patients and regurgitation in 5, while 2 patients had endocarditis. The operation was performed by a subcoronary technique in 9, root-inclusion technique in 3, and full root technique in 1 patient. RESULTS: Throughout the follow-up periods (with average follow-up period of 20.6 months), there was no hospital mortality, though there was one late death of unknown cause. The New York Heart Association class improved in all patients. The peak transvalvular gradient decreased from 18.4 +/- 9.8 to 12.6 +/- 9.6 mmHg, and the effective valve orifice area increased from 2.30 +/- 0.96 to 2.59 +/- 1.05 cm2 between the 1-month and the 6-month follow-up examinations. In patients with aortic regurgitation, the left ventricular end diastolic/end-systolic volume index significantly decreased from 147 +/- 36/62 +/ 19 to 73 +/- 26/33 +/- 14 ml/m2 at 1 month after the operation. The left ventricular mass index also significantly decreased from 189 +/- 26 to 143 +/- 30 g/m2 in patients with aortic regurgitation and from 171 +/- 28 to 144 +/- 30 g/m2 in those with aortic stenosis. CONCLUSIONS: Although long-term follow-up is required for further evaluation, the early results appeared to indicate that the Freestyle aortic bioprosthesis was suitable for elderly patients requiring aortic valve replacement. PMID- 10824476 TI - Pulmonary mucosa-associated lymphoid tissue lymphoma 8 years after resection of the same type of lymphoma of the liver. AB - A 64-year-old woman presented with hepatic and pulmonary tumors of mucosa associated lymphoid tissue lymphoma occurring 8 years apart. The present case carries the possibility of pulmonary metastasis of hepatic lymphoma or double primary lymphoma. Mucosa-associated lymphoid tissue lymphomas tend to develop in the extranodal primary organ, rarely developing systemically among extranodal organs as in our case. Our experience is useful in understanding the progress and outcome of mucosa-associated lymphoid tissue lymphoma. PMID- 10824475 TI - Minimally invasive direct coronary artery bypass grafting in a patient with brainstem infarction. AB - Brainstem infarction associated with the primitive trigeminal artery is rare. We describe the case of a 61-year-old man with an acute myocardial infarction as well as a brainstem infarction. The patient was referred for coronary artery bypass grafting. Minimally invasive direct coronary artery bypass grafting (left internal thoracic artery to the second diagonal branch anastomosis) could be safely performed 10 weeks after an episode of brainstem infarction. PMID- 10824477 TI - Tamponade after open-heart surgery with percutaneous cardiopulmonary support. AB - A 56-year-old man presented with late cardiac tamponade appearing on 9 postoperative day after weaning from percutaneous cardiopulmonary support. He had been referred to our hospital for congestive heart failure. He underwent aortic valve replacement and fell into postcardiotomy low output syndrome. He could not be weaned from extracorporeal circulation, and we had to use an intraaortic balloon pump and percutaneous cardiopulmonary support. On postoperative day 9, percutaneous cardiopulmonary support was successfully withdrawn without problems, but he showed signs of superior vena cava syndrome after the cannulas were removed. An echocardiogram also showed cardiac tamponade. When the wound was reopened, a lot of old clots had compressed the right atrium and, after clot removal, the patient's hemodynamic state improved markedly. It is important to be aware that percutaneous cardiopulmonary support may conceal hemodynamic deterioration due to cardiac tamponade and to take care that a patient does not experience hemodynamic deterioration after percutaneous cardiopulmonary support withdrawal. PMID- 10824478 TI - Chylothorax after thoracoscopic esophagectomy. AB - Among 30 patients who underwent thoracoscopic esophagectomy with lymphadenectomy for thoracic esophageal cancer, from July 1995 to May 1997, chylothorax developed in 2 patients (7%). In Case 1, the ligation of the thoracic duct under conventional right thoracotomy was performed on the 9th day after esophagectomy. After ligation, the pleural effusion was decreased, and the patient was discharged from hospital on the 25th day after the second operation. In Case 2, massive pleural effusion developed on the 10th day after esophagectomy (at 3 days after thoracic drainage tube was removed). The thoracic duct was ligated at the level just cranial to the diaphragm thoracoscopically on the 14th day after esophagectomy. The patient was discharged from hospital on the 30th day after the second operation. Injury to the thoracic duct due to a magnification effect of the view of scopic surgery remains a pitfall in thoracoscopic esophagectomy. But thoracoscopic ligation of thoracic duct was effective and safe for these two cases of chylothorax after esophagectomy. PMID- 10824479 TI - Difference in structural change in the Carpentier-Edwards pericardial valves implanted in the mitral and tricuspid positions. AB - We report a 29-year-old patient with prosthetic valve dysfunction with severe calcific stenosis in the mitral position but no structural change in the tricuspid position after mitral valve replacement and tricuspid valve supra annular implantation with same bioprostheses at the seven years before. The difference in structural change between the mitral position and the tricuspid position might be due mainly to the effect of mechanical stress on the cusps, rather than to any difference in serum calcium levels. However, some hormonal effect other than that of the parathyroid hormone on the systemic and pulmonary circulation might be related to the early progression in cusp calcification in the systemic circulation. PMID- 10824480 TI - Upper hemisternotomy as conversion from minimally-invasive coronary artery bypass grafting. AB - A seventy-year-old man was admitted to hospital for ischemic heart disease and abdominal aortic aneurysm. In the cardiac procedure, we employed a technique for conversion from minimally invasive coronary artery bypass grafting. This technique entailed cardiopulmonary bypass using standard instruments and technique, and the exposure for grafting was the same as for the simple minimally invasive coronary artery bypass grafting. Moreover, the incision we reported in this case was simply extendable even to a full sternotomy if necessary. PMID- 10824481 TI - Thoracic aortic aneurysm complicated with severe coronary arterial occlusive disease. AB - The patient was a 77-year-old female who had been treated medically for angina pectoris since 5 years ago. Expanded aneurysms in the distal aortic arch and in the descending thoracic aorta were seen during follow-up. She presented continuous back-pain at rest along with increasing size of the aneurysms despite antihypertensive therapies after admission. First, two saphenous vein grafts were anastomosed to the left anterior descending artery and obtuse marginal artery under beating heart. Next, the proximal portion of the left subclavian artery was clamped and divided. To this graft, the proximal ends of the coronary bypassed vein grafts were anastomosed and coronary perfusion was established and maintained until this artery was anastomosed to the aortic graft. Then, the aneurysms in the distal arch and descending thoracic aorta were excised and the aorta and its two pairs of intercostal arteries were reconstructed. The Postoperative course was uneventful with favorable cardiac function. PMID- 10824482 TI - Pulmonary sequestration associated with aspergillosis. AB - Pulmonary sequestration involves an abnormal pulmonary tissue separated from the normal pulmonary parenchyma, not connected to the tracheobronchial tree and supplied by a systemic artery. A case of intralobar pulmonary sequestration is presented. Case; a 49-year-old male was admitted to our hospital complaining of fever, cough and sputum production. Sputum culture resulted in a large growth of Aspergillus niger. Angiography showed an abnormal blood supply from the abdominal aorta to the right lower lobe. Right lower lobectomy was performed. The postoperative courses are uneventful. PMID- 10824483 TI - Posterior ventricular septal perforation in an 80-year-old man. AB - Ventricular septal perforation is a complication of acute myocardial infarction that is fatal unless treated surgically. Posterior ventricular septal perforation remains particularly difficult to repair successfully. We report an 80-year-old man with postinfarction posterior ventricular septal perforation who successfully underwent surgical repair using a technique similar to that described by Daggett et al. and was discharged in good health without postoperative septal shunt. PMID- 10824484 TI - Watershed infarction after combined coronary and axillobifemoral bypass surgery. AB - Watershed infarction sometimes causes a neurological disorder due to hypoperfusion of the brain during cardiac surgery. Here we report a case in which watershed infarction developed after combined coronary artery bypass and axillobifemoral bypass surgery. PMID- 10824485 TI - A retrospective survey of mucocutaneous manifestations of HIV infection in Malaysia: analysis of 182 cases. AB - A retrospective analysis of 182 HIV positive Malaysians was done in two centers, the University Hospital Kuala Lumpur (UHKL) and the General Hospital Kuala Lumpur (GHKL) from March 1997 to February 1998. Demographic and clinical data were analyzed. The analysis showed that 130 out of 182 patients had mucocutaneous disorders (71.4%). In the study there were 125 males (96.2%) and 5 females (3.8%). The majority of the patients were in the age group from 20 to 50 years. The patients who presented with mucocutaneous disease also had low CD4+ T lymphocyte counts, and most of them had AIDS defining illnesses. The number of cases with generalized hyperpigmentation was very high (35.7%), followed by papular eruptions (29.1%) and xerosis (27.5%). Seborrheic dermatitis was seen in 19.2% of the cases and psoriasis in 7.7%. The most common infections were oral candida 35.7%, tinea corporis and onychomycosis 9.9%, and herpes infection 4.3%. However, mucocutaneous manifestations of Kaposi's sarcoma were rare. The results suggested that mucocutaneous findings are useful clinical predictors of HIV infection or a sign of the presence of advanced HIV infection. PMID- 10824486 TI - A case of combined nevus: compound nevus and spindle cell Spitz nevus. AB - Combined nevi consisting of a Spitz nevus and an acquired nevus are unusual, and, to our knowledge, the combination of a spindle cell Spitz nevus and an overlying compound nevus has not been previously reported. We report a 17-year-old girl with a nodule on the left anterior lower extremity. The nodule was asymptomatic, firm, brown, symmetrical, dome-shaped, 8 mm in diameter, and not found with ulceration. Histological findings showed proliferation of spindle-shaped cells with an overlying compound nevus. The spindle-shaped cells were large, non pigmented, uniform in size and shape, with rare mitoses and without nuclear atypia, and arranged in a storiform pattern in thick collagen bundles. They stained positively for S-100 and negatively for HMB-45. The lesion was considered to be a new type of combined nevus consisting of a spindle cell Spitz nevus and an overlying compound nevus. PMID- 10824487 TI - Analysis of T-cell antigen receptor gamma chain gene rearrangement by polymerase chain reaction in combination with denaturing gradient gel electrophoresis in the differential diagnosis of cutaneous T-lymphoproliferative diseases. AB - A polymerase chain reaction (PCR)-based strategy has been developed for analysis of clonal rearrangement of the T-cell receptor gamma gene (TCR gamma) and was shown to be useful for detection of clonal T-cell populations. In this study, we performed PCR combined with denaturing gradient gel electrophoresis (DGGE) on fresh frozen biopsy samples from 16 patients with cutaneous T-lymphoproliferative diseases in whom a definite diagnosis was difficult to make on morphological and immunohistochemical grounds alone. Ages of the patients at biopsy ranged from 28 to 81 (median 62) years, and the subjects consisted of 8 men and 8 women. They presented with erythema on the extremities in 5 cases, trunk in 7, buttock in 2, and papules on the trunk and face in one case each. Clonal rearrangement of TCR gamma was observed in 3 of 16 cases. Clinical diagnoses of these three cases were mycosis fungoides, cutaneous invasion of adult T-cell leukemia (ATL), and large granular lymphocytic leukemia (LGL) of T-cell type, respectively, but they were histologically difficult to differentiate from reactive cutaneous T-cell proliferation. The skin lesions of the LGL case worsened, and this patient died two years after biopsy. Another patient with suspected mycosis fungoides in the plaque stage died due to dissemination of tumors 22 months after biopsy. The remaining one patient with ATL survived with cutaneous lesions for over four years. Clonality was not demonstrated in the remaining 13 cases, and their clinical courses were favorable. These findings showed that demonstration of clonal TCR gamma gene rearrangement using the PCR-DGGE method is very helpful for diagnosis of cutaneous T-cell neoplasms. PMID- 10824488 TI - Enhancement of adenovirus-mediated gene transfer into dermal fibroblasts in vitro and in vivo by polyethylene glycol 6000. AB - Gene therapy directed to the skin requires efficient transfer of the desired gene into cutaneous cells. In this study, we examined several chemical substances present in various ointments by enhancement of virus infectivity. The recombinant adenovirus vector, AxCALacZ, was used to infect dermal fibroblasts with some chemicals both in vitro and in vivo, and the expression of the LacZ gene was determined by X-Gal reaction. As the result, it was shown that PEG 6000 had the highest ability to enhance the exogenous gene expression in cultured fibroblasts with little toxicity. In vivo, it was also demonstrated that fibroblasts in mouse skin were efficiently gene transfer by adenovirus vector and 20% PEG 6000 treatment. These results suggest that this chemical treatment appears to be a simple, safe, convenient, and effective method for facilitating virus-mediated gene therapy in the skin. PMID- 10824489 TI - Herpes zoster infection complicated by motor paralysis. AB - We reviewed a total of 1,432 patients diagnosed with cutaneous herpes zoster at Hyogo College of Medicine Hospital between 1989 and 1997 for epidemiologic assessment and outcome in patients with zoster paralysis referred for rehabilitation. Of the 1,432 herpes zoster patients (624 males and 808 females, mean age 54.3 years), 12 were referred to our department of rehabilitation medicine for muscle weakness: one had myelitis, and eleven others had lower motor neuron damage. Except for one 43-year-old man with myelodysplastic syndrome, all the lower motor neuron deficit patients were over 60 years of age. Involved myotomes were more widespread than involved dermatomes in eight patients. Electromyography in four patients demonstrated denervation of involved muscles. Five patients experienced complete or near complete recovery from their muscle weakness. The muscle weakness related to herpes zoster was occasionally diagnosed by electromyography as motor neuron damage. Manifestations of motor neuron complications were not noticed but might in fact be more common than was the clinical muscle weakness. PMID- 10824490 TI - Effect of zinc supplementation on epidermal Langerhans' cells of elderly patients with decubital ulcers. AB - In the present study, Langerhans' cells (LCs) in the sacral epidermis, 8-10 cm from lesions of patients (mean age 71 years) with decubital ulcers, were compared ultrastructurally and morphometrically with those in the patients' own normal epidermis from the upper leg, before and after supplementation with 50 mg/day elementary zinc (in the form of a 220-mg tablet of Avazinc, administered once daily) for four months. Zinc intake resulted in from 80% to full healing of the decubital ulcers in the patients studied. The percentages of LCs were low in both perilesional sacral epidermis (2.07 +/- 0.71%) and in control leg epidermis (2.71 +/- 1.38%) before zinc supplementation and also afterward (2.12 +/- 0.16% and 2.59 +/- 0.88%, respectively). LCs demonstrated a more dendritic morphology after zinc supplementation: 68.15 +/- 9.28% and 77.0 +/- 3.45% of sacral and of control leg epidermal LCs, respectively, had dendrites before, and 91.52 +/- 3.43% and 84.15 +/- 3.64% of sacral and of control leg epidermal LCs, respectively, had dendrites after zinc supplementation. The LC section area in the sacral epidermis near the lesion as well as in the control leg epidermis and the number of LC granules in LC sections were not affected by zinc supplementation. The higher percentage of LCs having dendrites in the epidermis of patients with decubital ulcers after zinc supplementation may indicate that these LCs are in a more motile state, which might affect the healing process of the lesions. PMID- 10824491 TI - Sporotrichoid dermatosis caused by Mycobacterium abscessus from a public bath. AB - Infections caused by nontuberculous mycobacteria (NTM) are usually associated with immunocompromised states. More recently, however, NTM infections are being diagnosed with greater frequency in patients lacking traditional risk factors. However, cutaneous infection with rapidly growing mycobacteria is uncommon, and diagnosis may be difficult. Herein we present a case of sporotrichoid dermatosis on both forearms caused by Mycobacterium abscessus in a 34-year-old female (case 1). Mycobacterium abscesus was identified by culture as a colorless colony with rapid growth and by comparative sequence analysis of the rpoB gene. The patient was suspected to have been infected in a public bath in which she worked, it was located in a famous hot spring area in Korea. The condition was first noticed after she had been working in the bath for two years and after another employee (case 2) suffered similar lesions which had responded to treatment. The patient's skin lesions were successfully treated with anti-tuberculous drugs for six months. PMID- 10824492 TI - Palmoplantar pustulosis with pustulotic arthroosteitis involving temporal osteomyelitis and meningitis. AB - A 46-year-old man who had been suffering from palmoplantar pustulosis (PPP) for 3 years had anterior chest pain and left temporal pain from six months after the onset of his disease. A bone scan revealed abnormal uptake at the sternoclavicular joint and left temporal region. The head CT and MRI gave the diagnosis of temporal osteomyelitis with meningitis and myositis. His headache continued even after tonsillectomy and was effectively treated with cyclosporine A (3 mg/kg/day). Oral cyclosporine A was beneficial for the osteomyelitis and skin lesions. Sterile lytic bone lesions occurring most often at the sternocostoclavicular joint have been associated with PPP. However, there have been no reports of a PPP patient with temporal osteomyelytic involvement. PMID- 10824493 TI - Hyalinizing Spitz nevus. AB - A seventeen-year-old Korean girl had a reddish-brown papular lesion on the nose. Histopathologically, it proved to be a "hyalinizing Spitz nevus" with the characteristic features of a discohesive growth pattern of nevus cells and hyalinized stroma. Immunohistochemical stains showed positive reactivity of nevus cells with S-100 protein and Vimentin and negative stainings with HMB45, CD68, CEA and low molecular weight cytokeratin. Hyalinizing Spitz nevus may represent a variant in the spectrum of Spitz nevus. PMID- 10824494 TI - Two cases of multiple giant pilomatricoma. AB - Pilomatricoma is a skin appendage tumor manifestating as a firm, solitary lesion of the face and upper extremity. It generally measures 0.5 to 3.0 cm in diameter and is typically found in young people. We report a 29-year-old male and a 13 year-old girl who developed three and two protruding masses, respectively. The size of each tumor was larger than 5 cm. The histologic appearance showed sharply demarcated tumor lobules composed of eosinophilic shadow cells and basaloid cells. We describe two unusual cases of multiple giant pilomatricomas and review the literature, emphasizing the number and size. PMID- 10824495 TI - A case of adult T-cell lymphoma leukemia with hemophagocytic syndrome. AB - A 62-year-old Japanese woman was admitted to our clinic with virus-associated hemophagocytic syndrome (VAHS) in subcutaneous adult T-cell lymphoma leukemia (ATLL). Bone marrow aspiration showed hypocellularity, histiocytic hyperplasia, and hemophagocytosis. There was serological evidence of chronic cytomegalovirus (CMV) infection. The hemophagocytic syndrome (HPS) initially improved by some treatments, and the patient later experienced remission several times, but the CMV infection persisted. Most cases of non-tumorous HPS in adults are associated with viral or bacterial infection, and underlying diseases in non-tumorous HPS are mostly blood diseases, especially T-cell lymphoma (1, 2), but ATLL is a rare underlying disease in such cases. PMID- 10824496 TI - A case of pseudogout. AB - We report a 56-year-old Japanese male with a gout-like acute attack of synovitis and panniculitis in his right wrist without hyperuricemia. Calcium pyrophosphate dihydrate (CPPD) crystals were found in the synovial fluid of the affected joint. A diagnosis of pseudogout was made. The attack subsided two days after medication with 30 mg of predonisolon. Although this is a relatively common disease in elderly people, we rarely encounter such case reports in dermatological journals. PMID- 10824497 TI - Pedunculated lipofibroma. AB - Pedunculated lipofibroma is a relatively rare form of nevus lipomatosus superficialis. The lesions are large, slow growing, pedunculated tumors. Histologically, groups and strands of fat cells are found embedded among the collagen bundles of the dermis. We presented a woman diagnosed with a pedunculated lipofibroma clinically and histopathologically. PMID- 10824498 TI - Conceptual apraxia in probable Alzheimer's disease as demonstrated by the Florida Action Recall Test. AB - Patients with probable Alzheimer's disease (AD) often have difficulties associated with semantic knowledge. Therefore, conceptual apraxia, a defect of action semantics and mechanical knowledge, may be an early sign of this disease. The Florida Action Recall Test (FLART), developed to assess conceptual apraxia, consists of 45 line drawings of objects or scenes. The subject must imagine the proper tool to apply to each pictured object or scene and then pantomime its use. Twelve participants with Alzheimer's disease (NINCDS-ADRDA criteria) and 21 age- and education-matched controls were tested. Nine Alzheimer's disease participants scored below a 2-standard-deviation cutoff on conceptual accuracy, and the three who scored above the cutoff were beyond a 2-standard-deviation cutoff on completion time. The FLART appears to be a sensitive measure of conceptual apraxia in the early stages of Alzheimer's disease. PMID- 10824499 TI - Cognitive functioning after whiplash injury: a meta-analysis. AB - Complaints on cognitive functioning are often reported in patients suffering from whiplash syndrome, although objective neuropsychological test results do not always support these. In addition, radiological abnormalities and anatomical lesions are found only in a minority of these patients. This has led to a controversy about its existence in the literature. In this systematic review, the results of 22 neuropsychological studies on whiplash were quantitatively analyzed, focusing on working memory, attention, immediate and delayed recall, visuomotor tracking, and cognitive flexibility. Our findings suggest that a consistent overall pattern of cognitive dysfunction can be demonstrated after whiplash injury through neuropsychological testing, both compared to healthy and to asymptomatic controls. Six months after the accident, improvement is found in working memory, attention, immediate recall, and visuomotor tracking. The results are discussed in the light of recent findings on the effect of cerebral dysfunction, malingering, pain-related factors, and the role of coping strategies and posttraumatic stress on neuropsychological test performance. PMID- 10824500 TI - Personality change disorder in children and adolescents following traumatic brain injury. AB - The occurrence of personality change due to traumatic brain injury (PC), and its clinical and neuroimaging correlates were investigated. Ninety-four children, ages 5 through 14 at the time of hospitalization following traumatic brain injury (TBI; severe TBI N = 37; mild-moderate TBI N = 57), were assessed. Standardized psychiatric, adaptive functioning, cognitive functioning, family functioning, family psychiatric history, severity of injury, and neuroimaging assessments were conducted. The Neuropsychiatric Rating Schedule (NPRS) was used to establish a diagnosis of PC. Approximately 40% of consecutively hospitalized severe TBI participants had ongoing persistent PC an average of 2 years postinjury. An additional approximately 20% had a history of a remitted and more transient PC. PC occurred in 5% of mild-moderate TBI but was always transient. Interrater reliability for the diagnosis of PC was good (Kappa = .70). In severe TBI participants, persistent PC was significantly associated with severity of injury, particularly impaired consciousness over 100 hr, adaptive and intellectual functioning decrements, and concurrent diagnosis of secondary attention deficit hyperactivity disorder, but was not significantly related to any psychosocial adversity variables. These findings suggest that PC is a frequent diagnosis following severe TBI in children and adolescents, but is much less common following mild-moderate TBI. PMID- 10824501 TI - Comparison of the serial position effect in very mild Alzheimer's disease, mild Alzheimer's disease, and amnesia associated with electroconvulsive therapy. AB - Individuals given a series of words to memorize normally show better immediate recall for items from the beginning and end of the list than for mid-list items. This phenomenon, known as the serial position effect, is thought to reflect the concurrent contributions of secondary and primary memory, respectively, to recall performance. The present study compared the serial position effects produced on Trial 1 of the California Verbal Learning Test (CVLT) in mildly demented (N = 25; M MMSE = 20.0) and very mildly demented (N = 25; M MMSE = 25.5) patients with Alzheimer's disease (AD), and age- and education-matched normal control (NC) participants (N = 50). In addition, the serial position effects of the very mildly demented AD patients were compared to those of patients with a transient, circumscribed amnesia arising from a prescribed series of electroconvulsive therapy (ECT) treatments for the relief of depressive illness (N = 11). While the NC group exhibited the typical serial position effect, AD patients recalled significantly fewer words than NC participants overall, and exhibited a significantly reduced primacy effect (i.e., recall of the first 2 list items) with a normal recency effect (i.e., recall of the last 2 list items). Patients with circumscribed amnesia due to ECT were as impaired as the very mildly demented AD patients on most standard CVLT measures of learning and memory, but exhibited primacy and recency effects, which were within normal limits. These results suggest that a reduction in the primacy effect, but not the recency effect, is an early and ubiquitous feature of the memory impairment of AD. It is not, however, a necessary feature of all causes of memory impairment. PMID- 10824502 TI - Rehabilitation of executive functioning: an experimental-clinical validation of goal management training. AB - Two studies assessed the effects of a training procedure (Goal Management Training, GMT), derived from Duncan's theory of goal neglect, on disorganized behavior following TBI. In Study 1, patients with traumatic brain injury (TBI) were randomly assigned to brief trials of GMT or motor skills training. GMT, but not motor skills training, was associated with significant gains on everyday paper-and-pencil tasks designed to mimic tasks that are problematic for patients with goal neglect. In Study 2, GMT was applied in a postencephalitic patient seeking to improve her meal-preparation abilities. Both naturalistic observation and self-report measures revealed improved meal preparation performance following GMT. These studies provide both experimental and clinical support for the efficacy of GMT toward the treatment of executive functioning deficits that compromise independence in patients with brain damage. PMID- 10824503 TI - An unbalanced distribution of inputs across the hemispheres facilitates interhemispheric interaction. AB - In this study, we investigated 2 possible mechanisms by which interhemispheric interaction (IHI) might facilitate performance. Twenty university students performed 3- and 4-item versions of a less complex physical identity (PI) task in which they decided whether 2 letters were perceptually identical (e.g., 'A' and 'A') and a more complex name identity (NI) task in which they decided whether 2 letters had the same name (e.g., 'A' and 'a'). Consistent with prior work, IHI facilitated performance more for the relatively complex NI task than for the simpler PI task regardless of how many items were in the display. However, for each task IHI facilitated performance less in the 4-item displays than in the 3 item displays. These results indicate that IHI facilitates performance by allowing (1) a division of processing across the hemispheres, and (2) task relevant information to be processed by a hemisphere that receives a relatively light processing load. PMID- 10824504 TI - HIV-1, cocaine, and neuropsychological performance in African American men. AB - The purpose of this study was to examine the independent and interactive effects of HIV-1 serostatus and cocaine on neuropsychological (NP) performance in a sample of 237 gay and bisexual urban-dwelling African American men. Consistent with current evidence, it was expected that the greatest neuropsychological performance deficits would be evident (1) in the symptomatic seropositives (SSPs), especially in domains affected by HIV (i.e., memory and psychomotor speed), and on tests that are sensitive to subtle slowing; (2) in those who are recent and frequent cocaine abusers; and (3) in those who are both HIV seropositive and cocaine abusers. Multivariate analyses controlling for age and alcohol use confirmed expectations, with symptomatic seropositives (SSPs) evidencing significantly poorer psychomotor speed than the seronegatives (SNs), and slower reaction time and poorer nonverbal memory than the asymptomatic seropositives (ASPs). Moderate to heavy recent cocaine use was associated with slower psychomotor speed. However, contrary to expectations, no interaction of serostatus and cocaine was noted for any NP domain, and the expected serostatus and cocaine effects on verbal memory and frontal systems were not obtained. Level of alcohol consumption exacerbated the detrimental effects of HIV-1 on a computerized reaction time test which is especially sensitive to subtle slowing. This study provides one of the first descriptions of the neuropsychological effects of HIV-AIDS in a non-injection drug-using community sample of gay and bisexual African American men. PMID- 10824506 TI - Arousal response with aging: pupillographic study. AB - The performance of cognitive behaviors requires an activated, aroused cerebral cortex. Although studies have shown that there are decrements of cognitive functions in the elderly, changes in arousal with aging have not been fully studied. Our objective was to learn if there are attention-arousal changes associated with aging. Visual stimuli were presented to induce orienting responses or arousal reactions. Because changes in pupil size reflect changes in arousal, we recorded and compared pupillary responses of young and older normal participants using infared pupillography. During the 1 s that we recorded pupillary changes, we found major phases: a brief initial constriction (C1), then a maximal dilation (D1)--an arousal response, followed by constriction (C2), a habituation response. Although amplitude of these 3 phases was not different between the 2 groups, the interval between the D1 and C2 response was prolonged in the older group. Although the arousal response is not dramatically altered with aging, habituation appears to be delayed. PMID- 10824505 TI - Neuropsychiatric disturbance is associated with executive dysfunction in HIV-1 infection. AB - Prominent apathy and/or irritability are frequently observed among individuals infected with the human immunodeficiency virus (HIV). Although these symptoms often occur as part of a mood disorder, compelling evidence suggests that they may occur independently of depression in neurologic disease/disorder. The current study examined the prevalence of both apathy and irritability among a sample of HIV-infected individuals and explored the degree to which these neuropsychiatric (NP) phenomena were associated with performance on neuro-cognitive measures thought to be sensitive to the potential CNS effects of HIV-1. Clinician administered rating scales assessing apathy and irritability were administered to 65 HIV-seropositive (HIV+) and 21 HIV-seronegative (HIV-) participants who also completed a dual-task reaction time paradigm and the Stroop task. NP disturbance was significantly more prevalent among HIV+ participants compared with HIV- controls and was associated with specific neuro-cognitive deficits suggestive of executive dysfunction. Relative to both HIV- controls and to neuro psychiatrically intact HIV+ participants, those HIV+ individuals with evidence of prominent apathy and/or irritability showed deficits in dual-task, but not single task, performance and on the interference condition of the Stroop. Unexpectedly, NP disturbance did not show a robust relationship with HIV disease stage. These results suggest that the presence of prominent apathy and/or irritability among HIV+ individuals may signify greater HIV-associated CNS involvement. In HIV/AIDS, the disruption of frontal-subcortical circuits may be a common mechanism causing both executive dysfunction and NP disturbance. PMID- 10824507 TI - Functioning of the corpus callosum in children with early hydrocephalus. AB - The development and organization of the corpus callosum is described as well as the relationship between the timing of insults and the type of partial agenesis of the corpus callosum are discussed. Neuropathology and callosal damage associated with spina bifida meningomyelocele, aqueductal stenosis, and prematurity-IVH are outlined. Relationships between corpus callosum/whole brain ratios and cognitive functioning as well as interhemispheric transfer in children with these disorders are outlined. Shortcomings of current research and future directions are suggested. PMID- 10824508 TI - Reliable change formula query. PMID- 10824509 TI - [A morphological study of the kidney in the Japanese fetus with special reference to their external form, size and number of minor calix]. AB - There are few anatomical reports on the fetal kidney than on adult human and animal kidneys. This paper reports the size, weight, ratio of renal hilus, number of minor calix and number of renal lobe in Japanese fetal kidneys. Fifty kidneys of 25 fetuses (3, 8, 8 and 6 bodies in 5, 6, 7 and 8 months, respectively) were used in this study. The length, width, thickness and weight of kidneys were measured, and the renal lobes were counted. The relative size of the hilus was then calculated by the ratio of the area of hilus to that of the medial surface of the kidneys. Latex rubber was injected in the ureters, taken out of the calico pelvic system from the kidneys, and the number of minor calix in these specimens were counted. There were no significant sex difference and right and left difference in the kidney weight, ratio of renal hilus, number of minor calix and number of lobes. When these kidneys were observed at 5, 6, 7 and 8 months, the kidney weight and ratio of renal hilus were correlated with fetal age, but the numbers of lobe and minor calix were not. Comparing the results mentioned above with those of adult kidneys, it was shown that kidney weight and ratio of renal hilus area to the medial surface area of the kidneys gradually increased from fetus to adult. There was also a significant difference in the number of minor calix between adult and fetal kidneys. There was no correlation between the number of renal lobe and the number of minor calix. PMID- 10824510 TI - [Methods of phylogenetic reconstruction and comparative functional morphology]. AB - Cladistics is most prevalent method of phylogenetic reconstruction in the field of vertebrate paleontology, where it has been useful to overcome the weak points of previous evolutionary taxonomy and numerical phenetics. Its primary aim is to divide characters into two categories, old and new, and to bundle together taxonomic groups by their common derived characters (synapomorphies). In spite of such reasonable goals, the method also has limitations such as the premise for independency and equivalency of characters, deciding character polarity, and ignoring parallelism and convergence, as well as an inconsistency between phylogenetic positions and taxonomical names. It is important to understand the functional significance of the shape in question in order to eliminate mistakes in the quantitative treatment of morphology. Since the functions of various animals can be elucidated by mutual comparison of their morphologies, the author has termed this method "comparative functional morphology". One of the future trends for comparative morphology is a qualitative treatment of biological morphology that is more adapted to life, even in phylogenetic reconstructions. PMID- 10824511 TI - [Still-unknown determinants of the cranial shape]. AB - Although the shape of the neurocranium has long attracted the attention of anthropologists, the causes and mechanism of its formation have yet to be fully elucidated. At least from medieval times to the present, the superior view of the human skull has changed from an anteroposteriorly long shape to a short one. This is called "brachycephalization." Why does such a phenomenon occur? Does the cranial shape have a functional meaning? Main investigations, especially statistical, which have been carried out on these problems up to now, will be presented here. Reviewing the results of the investigations, they suggest, at least, that the shape of the neurocranium is correlated not only with the muscles of mastication and the jaws but also with the vertebrae, ribs and pelvis inside of the body, and, outside of the body, associated with environmental factors such as temperature, wealthiness, etc. In order to determine the concrete causes and mechanism for the formation of the cranium, it is emphasized in particular that not only human skeletal remains but also the paleoenvironmental data that are accompanied with them should intensively be collected in the future. PMID- 10824512 TI - [Morphological transformation of limb bones with growth]. AB - The development of the long limb bones consists of two developmental types, one for the length and the other for the thickness. Although the development of the length is due to the growth of the epiphyseal cartilage, the development of width is due to the addition of bone to the periosteal membranes. Growth of bones is influenced not only by chemical factor, such as hormone or vitamin, but also by physical factor such as electrical or mechanical stresses. Changes in the development of the limb bones caused by mechanical stress were studied. The limb bones, which belonged to a hydrocephalic patient who had been bedridden for 16 years, were measured. The lower limb bones had almost no mechanical stress such as weight or walking, due to his being bedridden for many years. He also could not exercise his upper extremity, hence mechanical stress to the upper limb bones was minimal. The circumference at the mid-shaft of each long bone was shorter than the average circumference of the same age. The thickness was also narrower, suggesting Wolff's law. However, no difference was found in length between the subject and the average. While mechanical stress was a factor that accelerated the thickness of the limb bones, stress did not have much influences on the length of the long limb bones. The length of the right clavicle was longer than that of the left due to the pressure of maintaining the same posture. In an experimental study of tail suspended rats showed that there was no significant difference in the length of hindlimb between the experimental group and control group, though a difference was found in the length of forelimb. The length of forelimb in the experimental group was significantly shorter than the control group due to weight pressure. This result suggests that physical pressure in the longitudinal direction is one of the factors that inhibits the growth of the length of bones and that mechanical stress is important for the morphology and the function of bones. Stress exerts different influence to accelerate or inhibit bones according to the direction. PMID- 10824513 TI - [Heritability of human skeletal morphology]. AB - The shape of human skeleton is determined by both genetic and environmental factors. In this report, I discussed the relative contribution of these two components in the osteometric traits by investigating 6 skeletal series affiliated to different social ranks. They were composed of 4 pedigrees, the Kanaseki family, the Tokugawa family (Shogun), the Makino family (Daimyo) and the Kuse family (Karo), and two control samples of unrelated individuals from the Edo period and recent times. The three familial samples, except the Kanaseki family, were the Japanese aristocrats of the Edo period. To estimate the family resemblance in skeletal morphology, I calculated Q-mode correlation coefficients (Q) and standardized Euclidean distances (D) for skeletal measurements of the relatives from 4 pedigrees and control samples of recent Japanese. Comparing Q and D between related and unrelated pairs, strong resemblances were detected among the related one. Family resemblance tended to reveal remarkable similarities in neuro-cranium, humerus, femur and tibia. Then, to determine the genetic contribution to these family resemblances, I estimated heritabilities (h2) for each measurement according to Falconer and Sjovold. High heritabilities were seen in skull measurements as mentioned by Sjovold. However, high heritabilities were also demonstrated in the epiphyseal part of the extremity bones. Environmental factors affecting the shape of human skeleton were analyzed by using canonical discriminant analysis for the measurements of the 6 skeletal series. Two environmental factors, social rank and the period that populations belong to, were detected from these analyses. Social rank seemed to be related with mechanical stress on the bone and the period with living condition such as nutrition and medical care. The influence by mechanical stress was remarkably recognized in such high mobility areas as facial cranium, forearm and leg. On the other hand, the influence on bones by living condition was more remarkable in the lower extremity than in the upper one. PMID- 10824514 TI - [Morpho-metrical features of the pelvis in standing posture]. AB - The morpho-metrical features of the human pelvis differ according to gender, particularly with regard to size and shape, and bipedal standing posture appears to have been a major determinant of pelvic structure. In standing posture, the three points of the right and left superior anterior iliac spines and pubic tubercle are in contact with the vertical frontal plane. In this situation, the superior anterior iliac spine is lower than the superior posterior iliac spine. From a lateral view, the vertical line from the body's center of gravity passes through the center of the hip joint or acetabulum. The anatomical or biomechanical problem has been whether the line of gravity passes through the promontory or the center of auricular surface. To clarify this point, the three points of the acetabular center, promontory and auricular center were examined as to how they are positioned in relation to each other and the distance of each on the vertical frontal plane. Those measurements are as followed: A) distance between vertical frontal plane and promontory, B) distance between vertical frontal plane and anterior margin of auricular surface, C) distance between vertical frontal plane and mid-point of auricular surface computed from (B + D)/2, D) distance between vertical frontal plane and posterior margin of auricular surface, E) distance between vertical frontal plane and mid-point of acetabulum, F) inclination angle of the pelvis. From the results, the acetabular center and promontory are in alignment with the vertical line. The auricular center is positioned further back than the acetabular center or promontory. In this situation, the mean angle of inclination of the pelvis was found to be 63 degrees in 16 Japanese male specimens. The means of the other Japanese populations are into range of 63 degrees to 66 degrees in the males. If the auricular center coincides with the acetabular center, it is possible the angle of inclination of the pelvis would exceed 63 degrees. The frontal plane in contact with the right and left superior anterior iliac spines and pubic tubercle is not vertical. The morphological features clarified in the present study are that the promontory and acetabular center are in vertical alignment when viewed laterally and that the angle of inclination of the pelvis is 63 degrees. PMID- 10824515 TI - [Growth and developmental patterns in Neandertals]. AB - Growth and development in Neandertals have been focused on in terms of the appearance of the human growth pattern in hominid evolution. Recently, more attention has been drawn to the aspects of growth in human evolution, although their significance was recognized from long before. One recently refined method is to use the perikymata counts on the enamel surface, which provide an absolute duration time of tooth crown development and therefore make possible comparisons in timings of developmental events. Another is to evaluate the relative timing and speed of developmental events from teeth, skull, and postcranial bones. In general, the hominid growth pattern would change from ape-like to human-like during the evolutionary process. It is still uncertain, however, when and where the exact event happened and the developmental pattern changed. As for the growth in Neandertals, it has been suggested that they would have grown relatively slowly in their childhood as modern humans do, while the appearance of the adolescent growth spurt and the duration time of their adolescence or childhood period are still obscure. More concrete and precise data are needed for a reasonable understanding of evolution of hominid growth for both modern humans and fossil hominids, in terms of: 1. more data of individual morphological traits applicable to fossil hominids, which increase the number of available fossil specimens; 2. variation of the traits in modern and archeological specimens, particularly in relation to environmental stress; 3. interaction or interrelationship between dental, cranial and postcranial development. PMID- 10824516 TI - [Cranial nonmetric variation of Yayoi people in the Kyushu District]. AB - The Yayoi people in Kyushu and Yamaguchi area are generally classified by metrical analyses mainly into the Yayoi people in the northern Kyushu and Yamaguchi area who are regarded as migrants from the Asian Continent and their posterity and the Yayoi people in the northwestern Kyushu who are regarded as having inherited the characteristics of the Jomon people. Such classification is verified by the analysis with the 22 traits in the cranial nonmetric variation. Of the 22 traits, supraorbital foramen, transverse zygomatic suture vestige, biasterionic suture vestige, jugular foramen bridging, hypoglossal foramen bridging, pterygospinous foramen, mylohyoid bridging and tympanic dehiscence are particularly important as the traits to classify two types of Yayoi peoples. The analysis by the C. A. B. Smith's Mean Measure of Divergence (MMD) suggests that out of the migrant Yayoi peoples, the very Yayoi people who are closely related to the formation of the modern Japanese are the ones in the northern Kyushu area, not the ones in the Doigahama site. Also, it is assumed that the appearance and disappearance of cranial nonmetric variations is affected by genetic elements, because the incidencies of cranial nonmetric variations is largely different between two types of Yayoi peoples in infants like in adults. Lastly the cranial nonmetric variation in the people of the period equivalent to Jomon and Yayoi in the Okinawa district, and in the Kofun people in southern Kyushu area, was briefly introduced. PMID- 10824517 TI - Signs of hope. PMID- 10824518 TI - Future directions for IPPNW. International Physicians for the Prevention of Nuclear War. AB - This paper is offered as my personal reflections for future directions of International Physicians for the Prevention of Nuclear War. The deep discussions that surrounded the war in Kosovo and Serbia have led me to examine some of the issues that confront IPPNW today in a different light. Rather than cataloguing all the work of IPPNW and the future steps in each, I focus on areas where we have a new opportunity, a new challenge, or a difficult decision to make in choosing a strategic direction for IPPNW. After a brief summary of our history and principles, I address the new challenges within IPPNW's triangle of concern- disarmament, environment and development. I then outline a series of strategies that have been proposed to advance our goals. Finally, I raise other issues that seem to warrant further discussion and policy development. PMID- 10824519 TI - Castles or boarding-houses: a new concept of security. AB - Current concepts of security, even if valid in the past, are inappropriate now. We need a new concept of security based on co-operation and interdependence, not military (and especially nuclear) confrontation. The International Court of Justice Advisory Opinion on the legality of nuclear weapons, international concern on the dangers of the arms trade, the landmines campaign, and statements by retired military leaders, are all signs of hope. But others, including non governmental organizations, must also learn to co-operate. The Year of the Culture of Peace is an opportunity to make co-operation more effective and, particularly in schools, to make the United Nations better known and better appreciated. PMID- 10824520 TI - A fast track to zero nuclear weapons: the Middle Powers Initiative and the New Agenda Coalition. AB - The Middle Powers Initiative is a network of international citizen organizations working to encourage the nuclear weapon states and their influential allies to move rapidly to eliminate nuclear weapons via practical steps including a Nuclear Weapons Convention. The New Agenda Coalition is a group of middle-ranking nations whose governments have also called for the early elimination of nuclear weapons via similar steps. The work of MPI and NAC at the United Nations and elsewhere is described, and their impact on NATO nuclear weapons policy discussed. PMID- 10824521 TI - Old dogs or new tricks: chemical warfare at the millennium. AB - There were about 50,000 deaths from chemical agents in the First World War and 7,000 deaths in the Kurdish population of Iraq from mustard gas and nerve agents. Signatories to the 1925 Geneva Protocol forswore first use of chemical warfare, but were allowed to maintain stockpiles, and there were no sanctions against use. The Chemical Weapons Convention (CWC), which entered into force in 1997, establishes timetables for declaration of current and past CW activity and destruction of stocks. The Convention allows inspection of suspect sites. The Organisation for the Prohibition of Chemical Weapons has been established to give effect to the CWC, which, as of June 1999, has 125 States Parties. Progress in reporting and destroying CW agents is described. It is essential that participation in the CWC becomes universal, but until this is the case monitoring by other agencies will be needed. PMID- 10824522 TI - Preventing deliberate disease. AB - The danger from deliberate disease as a weapon of war--biological weapons--is examined and it is concluded that biological weapons currently pose the greatest danger of all weapons of mass destruction. The ongoing work of the Ad Hoc Group negotiating a Protocol to strengthen the effectiveness and improve the implementation of the Biological and Toxins Weapons Convention is analysed. The Protocol is nearing completion and contains provisions for declarations of the most relevant facilities, for declaration follow up procedures including infrequent randomly-selected visits to declared sites, for investigations of non compliance concerns as well as measures to promote international co-operation for peaceful purposes in microbiology and biotechnology. It is concluded that an efficient and effective Protocol is achievable within the coming year. PMID- 10824523 TI - 'To save succeeding generations from the scourge of war': the role of the International Court of Justice. AB - The International Court of Justice is the judicial arm of the United Nations. Its provenance and role in keeping international peace and security is described. Its role in settling disputes between states and giving advice to the UN organs and specialized agencies is illustrated from a series of past cases where it has contributed to the maintenance or restoration of international peace and security. Possible limitations to its contributions, arising from the need for consent to its jurisdiction, and from the process of international law are discussed, and it is concluded that decisions under international law are for realization of values. Finally, its relationship with the UN Security Council is examined. PMID- 10824524 TI - An optimum world population. AB - The optimum population of the world is the one that is most likely to make the option of a good quality of life available to everyone everywhere, both now and in the future. Establishing a consensus about the size of such a population would be an important step towards achieving it. Estimates of an optimum involve three main steps. First, estimate the maximum (carrying capacity) assuming a specified lifestyle. The main criteria are the maintenance of biodiversity, the availability of freshwater, and the availability of land--for agriculture, forestry and artificial systems but above all for the conversion of energy. (In applying the criteria, there are always two questions to ask: 'What is the maximum amount of consumption that the biosphere can stand?' and 'What is an adequate share of such consumption per person?') Second, convert the maximum (two to three billion) into an optimum by applying a far wider range of criteria, including personal liberty, mobility, recreation and political representation. Third, consider just two criteria (economies of scale and technological innovation) in order to ensure that the optimum (one to two billion) has not fallen below the minimum (half to one billion). The estimates are so low because of the need for a huge increase in median per capita consumption if everyone is to have the option of an adequate material standard of living. Opinion-formers are likely not to take much notice of such estimates, but it is probable that minds will be concentrated by an energy shock some time during the next decade. Achieving an optimum world population will not solve the world's major problems, but it would make them solvable. PMID- 10824525 TI - Carson's syndrome: a major threat to the public's health in the twenty-first century. AB - Our environment provides essential support for our health, but is subject to degradation from human activities. This in turn has adverse effects on health. The name 'Carson's Syndrome' is proposed to characterize severe non-accidental planetary abuse. A specific example, overproduction of carbon dioxide (CO2), is described and illustrated by the concept of a 'footprint' for the CO2 production of an individual or group. Transport is responsible for 40% of CO2 production in developed countries. A more rational transport policy, with particular encouragement of walking and cycling, would both reduce CO2 output and benefit our health, especially in reducing cardiovascular disease. PMID- 10824526 TI - Feeding the world in the new millennium. AB - Food production per head in the world as a whole has begun to level off in the last decade, while the world population continues to grow, risking malnutrition, perhaps even starvation, civil unrest and environmental damage. Very little more land suitable for agriculture is available, and the factors behind the 'green revolution' may not produce further increases. Genetically modified crops offer the possibility of increased yields, but also present major problems. In developing countries, where yields are well below what is theoretically possible, the best approach may be better management by small farmers through improvements in their traditional methods. Much more financial support for and research in agriculture is needed, together with more equitable distribution of existing production and an end to trade practices designed to make the rich richer. PMID- 10824528 TI - Inducible genomic instability: new insights into the biological effects of ionizing radiation. AB - The biological consequences of exposure to ionizing radiation include gene mutation, chromosome aberrations, malignant transformation and cell death. These effects are attributed to the production of irreversible damage at the time of the irradiation. However, there is now considerable evidence that cells that have survived irradiation may produce descendants in which a high frequency of de novo chromosome aberrations and gene mutations arise or in which there is an enhanced death rate. These delayed effects are manifestations of an induced genomic instability. All the various delayed effects are induced at very high frequency and are unlikely to be due to conventional mutational changes. At present little is understood of the processes involved in the initiation of inducible instabilities and in the maintenance and transmission of the phenotype over many generations of cell replication. However, it is becoming evident that the expression of inducible instability has a strong dependence on the type of radiation exposure, the cell type irradiated and the 'genetic predisposition' of the irradiated cell. Radiation-induced genomic instability presents a major challenge to conventional radiobiology concepts and has important implications for mechanistic studies of radiation action particularly in the context of radiogenic malignancy. PMID- 10824527 TI - Global action to prevent war: a programme for government and grassroots efforts to stop war, genocide and other forms of deadly conflict. AB - At the end of history's bloodiest century and the outset of a new millennium, we have an opportunity to fulfil one of humanity's oldest dreams: making the world largely free of war. Global changes make this goal achievable. Nuclear weapons have shown the folly of war. For the first time, there is no war and no immediate prospect of war among the main military powers. For the first time, many proven measures to prevent armed conflict, distilled in the crucible of this century's wars, are available. If systematically applied, these measures can sharply decrease the frequency and violence of war, genocide, and other forms of deadly conflict. To seize the opportunity, nations should adopt a comprehensive programme to reduce conventional armaments and armed conflict. This programme will complement and strengthen efforts to eliminate nuclear arms. To assure its ongoing worldwide implementation, the conventional reduction programme should be placed in a treaty framework. We propose a four-phased process, with three treaties, each lasting five to ten years, to lay the groundwork for the fourth treaty, which will establish a permanent international security system. The main objectives of the treaties are to achieve: 1. A verified commitment to provide full transparency on conventional armed forces and military spending, not to increase forces during negotiations on arms reductions, and to increase the resources allocated to multilateral conflict prevention and peacekeeping. 2. Substantial worldwide cuts in national armed forces and military spending and further strengthening of United Nations and regional peacekeeping and peace enforcement capabilities. 3. A trial of a watershed commitment by participating nations, including the major powers, not to deploy their armed forces beyond national borders except in a multilateral action under UN or regional auspices. 4. A permanent transfer to the UN and regional security organizations of the authority and capability for armed intervention to prevent or end war, accompanied by further substantial cuts in national armed forces and increases in UN and regional forces. This programme offers many valuable features: a global framework for conventional forces that parallels the nuclear Non-Proliferation Treaty; a verified no-increase commitment for national armed forces based on full data exchange; a commitment to undertake prescribed confidence-building measures, including limits on force activities and deployments; a commitment to a specified plan for increased funding of UN and regional peacekeeping capabilities; a commitment to strengthen international legal institutions; and after a trial period, a lasting commitment by each participant not to unilaterally deploy its armed forces beyond its borders, but instead to give the responsibility for peacekeeping and peace enforcement to international institutions. This programme of phased steps to reduce armed forces and strengthen peacekeeping institutions will make war rare. It will foster the spread of zones of peace like those in North America and Western Europe where, after centuries of violence, international and civil war have given way to the peaceful settlement of disputes. PMID- 10824529 TI - Wounds, weapons and the doctor. PMID- 10824530 TI - [Clinical application of 133-Xe ventilation scintigraphy]. AB - Xenon-133 ventilation scintigraphy provides three sets of scintigrams: wash-in, equilibrium, and washout images. Krypton-81m or 99mTc-Technegas can be used as alternatives depending on their physical characteristics and the purpose of scintigraphy. Xenon-133 scintigraphy is compatible with SPECT when a multi detector gamma camera is used. Ventilation images are analyzed by the height/area method to calculate regional lung volume and the mean transit time of the posterior planar view. If perfusion scintigraphy is done during the same examination, a ventilation/perfusion ratio image is also obtained. Factor analysis is applicable to the washout phase and permits a more detailed depiction of uneven ventilation. Series of consecutive SPECT during washout (dynamic SPECT) visualize ventilation in cross section and provide three-dimensional images of regional lung volume and poorly ventilated areas. A major indication for 133Xe scintigraphy is acute pulmonary thromboembolism (PTE) when used in combination with perfusion scintigraphy. The scintigraphic diagnosis of PTE is not inferior to that of contrast-enhanced helical CT, although the latter was initially reported to have higher sensitivity and specificity than scintigraphy. Other important indications are differentiation between primary pulmonary hypertension and chronic pulmonary thromboembolism, and the prediction of postoperative pulmonary function. Ongoing trials are attempting to create functional images of ventilation by CT or MRI, both of which may become rivals of scintigraphy because of their higher image resolution. PMID- 10824531 TI - [99mTc-Technegas]. AB - 99mTc-Technegas (Technegas) scintigraphy, including planar and SPECT images, was reviewed in patients who mainly had pulmonary emphysema. The Technegas images showed different degrees of change, from areas of heterogeneity to hot spots or defects. Comparison of planar and SPECT Technegas images revealed that more detailed findings were shown by SPECT than by planar images in mild cases. In more severe cases, the findings of SPECT and planar images were equivalent. In advanced stages, SPECT images should be excluded in favor of planar images. Technegas SPECT could demonstrate ventilation impairment more easily than 133Xe gas dynamic SPECT. The optical average score of Technegas in whole lung correlated well with forced expiratory volume in one second (FEV1.0) and FEV1.0/forced vital capacity. Technegas could assess ventilation impairment in pulmonary emphysema more easily than CT, especially in the upper lung field. Technegas was useful for the assessment not only of pulmonary emphysema but also of other diseases. For the quantitative evaluation of Technegas, some investigators study three-dimensional fractal analysis or mean voxel values of the lung. These quantitative analyses are useful for classifying clinical stage and comparing cases. PMID- 10824532 TI - [Comparison of whole-body MR imaging and bone scintigraphy in the detection of bone metastases from breast cancer]. AB - The aim of this study was to investigate the usefulness of whole body MR imaging (WB-MRI) in the detection of bone metastases from breast cancer and to compare the results with those from bone scintigraphy. In 21 patients with suspected bone metastasis from breast cancer, both bone scintigraphy and WB-MRI were performed. With WB-MRI, coronal images were obtained using a body coil in an FOV of 48 cm, and sequences of fast short TI inversion recovery (STIR) and gadolinium-enhanced fast spoiled GRASS (SPGR) were used in three parts: from the head to the thorax, the abdomen to the pelvis, and the lower extremities. Of the total 105 metastatic bone lesions, 65 (61.9%) were detected by bone scintigraphy, 98 (93.3%) by fast STIR, and 74 (70.5%) by fast SPGR. Thus, the detection of bone metastases by WB MRI was excellent. However, detectability in the ribs was lower for WB-MRI than for bone scintigraphy. Contrast-enhanced MRI was useful in the differentiation of osteosclerotic lesions, in which high signal intensity is rare, pleural effusion, which has high signal intensity on STIR, and bone metastatic lesions. In conclusion, WB-MRI showed high reliability in the detection of bone metastatic lesions from breast cancer. PMID- 10824533 TI - [Long-term results of TIPS]. AB - The long-term results of transjugular intrahepatic portosystemic shunt(TIPS) were analyzed in 48 patients between February 1992 and December 1998, for treatment of esophageal varices and ascites caused by portal hypertension. The procedure was successful in 42 patients. Portal vein pressure was reduced from 29 mmHg +/- 7.9 to 19 mmHg +/- 7.3 immediately after TIPS and to 17 mmHg +/- 6.5 one year later. Varices were controlled in 82% (23/28) of patients after one year and in 78% (7/9) of patients after five years. Ascites disappeared or decreased in 79% (11/14) after one year. Primary shunt patency was 12%, with a secondary patency rate of 79% after five years. The overall incidence of new or worsened encephalopathy was 31% (13/42). Long-term follow-up showed one-, three-, and five year survival rates of 82%, 68%, and 59%, respectively. TIPS is an effective means of lowering portal hypertension, and it controls varices and ascites with little risk of hepatic encephalopathy. PMID- 10824534 TI - [Radiation dose measurement of the patient in interventional radiology using a transmission ionization chamber]. AB - We propose a method to estimate patient radiation dose in radiologically guided interventional procedures using a transmission ionization chamber. A typical transarterial embolization (TAE) procedure for hepatocellular carcinoma was simulated, including 30 minutes of fluoroscopy and five series of DSA, each with appropriate collimation. The dose-area product was divided by the area and compared with values from a standard dosimeter placed in the center of the radiation entrance, to obtain a conversion factor. In this way, the entrance skin dose can be estimated immediately after the procedure by simply multiplying the value by the conversion factor, if the procedure roughly conforms to the simulated model. The average entrance skin dose of 33 patients who recently underwent TAE for HCC was found to be 0.66 (0.19-1.75) Gy. This technique can be applied to other areas of IVR and may help to reduce patient exposure to radiation. PMID- 10824535 TI - [A study of dose distribution of high energy X-ray in lung measured by film method]. AB - Measurement was carried out for 6 MV X-rays using an inhomogeneity phantom with films. The films were set parallel or perpendicular to the beam axis. Measurement data were compared with values obtained by a JARP dosimeter, to assess the accuracy of evaluation. 1) The data measured by perpendicular films showed underestimation but were more accurate than those measured by parallel films when compared with the JARP method. 2) The difference in measurement between the perpendicular films with multi-layers and a single layer was 0.4%. 3) The difference in measurement between packed and unpacked films was 1.5%. The former overestimated values, but they were closer to the JARP values than those of unpacked film. 4) In comparison with the JARP method, accuracy decreased in the following order: perpendicular packed film with multiple layers, perpendicular packed film with a single layer, perpendicular unpacked film with multiple layers, perpendicular unpacked film with a single layer, parallel film. 5) The values measured by films in the inhomogeneity phantom were closer to those measured by the JARP method than values measured in a water-equivalent phantom. The reason was that there was less scatter in the lung of the inhomogeneity phantom than in the water-equivalent phantom. PMID- 10824537 TI - [Histogenosis and malignant progression of pulmonary carcinoma] [In Process Citation] PMID- 10824536 TI - [Development of a multi-layer ion chamber for measurement of depth dose distributions of heavy-ion therapeutic beam for individual patients]. AB - In heavy-ion radiotherapy, an accelerated beam is modified to realize a desired dose distribution in patients. The setup of the beam-modifying devices in the irradiation system is changed according to the patient, and it is important to check the depth dose distributions in the patient. In order to measure dose distributions realized by an irradiation system for heavy-ion radiotherapy, a multi-layer ionization chamber(MLIC) was developed. The MLIC consists of 64 ionization chambers, which are stacked mutually. The interval between each ionization chamber is about 4.1 mm water. There are signal and high voltage plates in the MILC, which are used as electrodes of the ionization chambers and phantom. Depth dose distribution from 5.09 mm to 261.92 mm water can be measured in about 30 seconds using this MLIC. Thus, it is possible to check beam quality in a short amount of time. PMID- 10824538 TI - [Advances in diagnosis and treatment of lung cancer]. AB - Advances in diagnostic methods and treatment modalities for lung cancer in the recent years were reviewed. Computed tomograph and other lung imaging devices facilitated early detection of small nodule, or diagnosis of extension of cancer. In addition to bronchofiberscopy, fluorescence endoscopy, tracheo-bronchial stenting, or photodynamic therapy brought marked advances in diagnosis as well as treatment of lung cancer at hilar and peripheral regions. Most of the comparative study to evaluate postoperative adjuvant therapy showed negative results. However, it is expected that induction therapy will improve surgical result of advanced lung cancer. Surgery for lung cancer showed great improvements in terms of operative mortality, operative methods (bronchoplastic surgery, combined resection of adjacent organ invasion, extensive lymph node dissection, or limited lung resection) and surgical outcome. PMID- 10824539 TI - [The outline of the general rule for clinical and pathological record of lung cancer]. AB - In 1999, the Japan Lung Cancer Society published the fifth Japanese edition of the General Rule for Clinical and Pathological Record of Lung Cancer. According to the new UICC's TNM classification revised in 1997, the TNM classification of this General Rule was modified. The guide to taking good conventional and high resolution chest computed tomogram was showed. The types of bronchoscopic findings was simplified. In operative procedures, curability of surgical resection was divided into only two groups: complete resection and incomplete resection. Although WHO/IASLC histological classification was revised in 1999, the histological classification hasn't been modified yet in this new edition of General rule. It will be changed in the next version. PMID- 10824540 TI - [Epidemiology of lung cancer in Japan]. AB - Lung cancer is the most common cause of cancer-related death in both men and women. There are over 51,000 deaths annually from this disease in Japan. The characteristics of lung cancer have changed for several decades. The incidence of the elderly, adenocarcinoma in histology, and stage I disease continues to increase from year to year. In the 1970s, six randomized trials were carried out to evaluate lung cancer screening in the U.S. and Europe. These studies did not contribute to decrease death rate. They concluded there is no evidence to recommend against lung cancer screening. On the other hand, in Japan, several trials by case-control study demonstrated that chest x-ray screening is associated with earlier detection and improved survival. The conclusions are controversial in two studies. Further investigation will be needed to define the benefit of cancer screening. PMID- 10824541 TI - [Molecular pathogenesis of lung cancer]. AB - Lung cancer is the largest cancer killer of men and women in the world. In addition to the progress made from antismoking primary prevention measures, new tools to help treat patients with lung cancer are emerging from the rapid advances in knowledge of the molecular pathogenesis of lung cancer. These tools include molecular and cellular biology and are starting to provide an insight into how the tumor cells, by altering oncogenes and tumor suppressor genes, achieves growth advantage, uncontrolled proliferation and metastatic behavior via disruption of key cell-cycle regulators and signal transduction cascades. These tools are being translated into clinical strategies to complement surgery, radiotherapy, and chemotherapy and also to assist in primary and secondary prevention efforts. From the current knowledge of the molecular pathogenesis of lung cancer we know that respiratory epithelial cells require many genetic alterations to become invasive and metastatic cancer. PMID- 10824542 TI - [Causative agents for lung carcinogenesis]. AB - Lung cancer is a leading cause of malignancy-related death in Japan, and its incidence is still rising steeply. Various factors, including cigarette smoking, asbestos, and diet, have been reported to correlate with the development of lung cancer. Of these factors, cigarette smoking is believed as the major carcinogen for lung cancer. Recent studies indicate that cigarette smoke carcinogens cause genetic damages at both oncogenes(K-ras) and tumor suppressor genes(p53) of lung cancer, and hence initiate and promote the development of lung cancer. This article reviews recent advances in the molecular mechanisms of lung cancer carcinogenesis. PMID- 10824543 TI - [Growth factor and signal transduction in lung cancer cell]. AB - Growth factors are molecules that participate in the control of cell proliferation. They require specific receptors on the target cell and intracellular signaling pathways to transmit the stimulus to the nucleus. Growth factors can stimulate or inhibit cell division. Most of the factors implicated in lung cancer growth are thought to act through positive feedback loops in which factors secreted by the cancer cells bind to receptors on their own surfaces(autocrine stimulation) or those of neighboring cells (paracrine stimulation). Abnormal expression of growth factors, their receptors, or components of their signaling pathways can result in the unrestrained growth of cancer. In this review, we described outlines of the growth factors, their receptors, signal transduction pathways, as well as their clinical applications. PMID- 10824544 TI - [Deregulation of cell cycle control in lung cancer]. AB - Studies on cell cycle regulation and cancer genetics have revealed that multiple cell cycle regulatory proteins play key roles in oncogenesis. These can be categorized in three sets. First; p16INK4-Cyclin D1-RB pathway, which controls G1 to S progression of the cell cycle, second; p53 pathway, which is involved in DNA damage repair, and third; p27KIP1 CDK inhibitor, a negative regulator of cell cycle, and decreased expression of which has been correlated to poor prognosis in cancer patients. Among these, p16INK4, RB and p53 are tumor suppressor genes, and p27 has been pointed out to be haplo-insufficient for tumor suppression. Involvement of these cell cycle regulatory proteins in lung cancer will be discussed. PMID- 10824545 TI - [Molecular biological approaches to lung cancer]. AB - Advances in lung cancer research have come from modern technology, allowing the establishment of lung cancer cell lines and experimental model systems, recombinant DNA technology, robust biological and biochemical functional assays in addition to classical methods such as immunohistochemistry. Such work indicates that the aberrant activation of protooncogenes and inactivation of tumor suppressor genes are fundamental processes for lung carcinogenesis. Further improvements such as DNA array technology will continue to unravel the genetic and epigenetic defects in a far more rapid and comprehensive way that was previously possible. This knowledge will lead to a genetic characterization of lung cancer cells and its specific subtypes, which will hopefully translate to new diagnostic and therapeutic gene based strategies. This review focuses on molecular biological techniques that have been used on lung cancer research and discusses future technological approaches. PMID- 10824546 TI - [Conquering drug resistance in lung cancer]. AB - The emergence of drug resistance of the tumors is the critical problem in lung cancer. The multifactorial mechanisms of cisplatin resistance are major problems. The reversal agents against MDR have been evaluated clinically. Increased adverse effects and modulation of pharmacokinetics in combined with cytotoxic drugs are the problems in clinical setting. The new ABC transporters related with MDR were cloned and characterized. Modulators for these transporters could be evaluated their clinical efficacy. High-frequent mutations of beta-tubulin in clinical samples are reported as a predictive marker for paclitaxel. Target-based drugs might conquer the resistance to conventional drugs. Prediction of the drug sensitivity and resistance in each patient will be expected using high-throghput gene screening system. PMID- 10824547 TI - [Gene therapy for lung cancer treatment]. AB - Gene therapy is a promising strategy for cancer treatment. The strategy is categorized to some fields based on the anti-cancer mechanism. p53 gene therapy and antisense therapy exert the cytotoxic effect on cancer cells by apoptosis induction. In suicide gene therapy, expressed suicide gene product kills the cancer cells in combination with prodrug which metabolite is cytotoxic agent. Oncolytic virus replicates in the tumor selectively and destroys the cancer cells. Immunogene therapy potentiates the antigenicity of cancer cells or activate the immune effector cells. In antiangiogenesis gene therapy, neovascularization is inhibited with antiangiogenic molecule gene such as angiostatin. On the other hand, bone marrow stem cells are protected from high dose anti-cancer agents by the transfer of multi-drug resistant gene. PMID- 10824548 TI - [Diagnosis--radiologic imaging]. AB - One of the most important recent advances in imaging technology for the diagnosis of lung cancer is CT consisting of high-resolution and helical volumetric techniques. CT has made a detailed morphological analysis correlated with histopathology as well as the detection of small peripheral lung cancers possible. MRI still remains as a complementary role to CT, but MR angiography using fast gradient-echo technique has achieved a great progress. FDG-PET is the most encouraging development in nuclear medicine and is vigorously evaluated for its cost-effective clinical value in differentiating benign and malignant nodules as well as in staging of lung cancer. In chest radiography, the development of computer-aided diagnosis is underway and seems promising. PMID- 10824549 TI - [Bronchofiberscopy]. AB - New diagnostic modalities have been used in conjunction with endoscopy for early detection of lung cancer. Videoendoscope is routinely used instead of fiberoptic bronchoscope. Fluorescence diagnosis has been proved to be useful in detecting subtle lesions which might be invisible by conventional endoscopy in central airway. Also, a number of small peripheral lesions has increased by the helical CT. CT guided transbronchial lung as well as needle cytology are indicated for definitive diagnosis of such lesions. Endobronchial Ultrasonography is employed to evaluate the depth of cancer invasion of the bronchus and lymph node swelling around the bronchus. It should be helpful in staging of lung cancer and selecting therapy. PMID- 10824550 TI - [Pathological diagnosis--practical treatment of surgical specimens with lung cancer]. AB - The importance of gross examination before histology of the surgically removed lung cancers was stressed by depicting the practical methods of formalin filling fixation, slicing the specimens, drawing and noting the findings, photographing, and cutting the tissue blocks. On gross examination of the sliced specimens it was guided that besides the pathological features of cancers themselves, secondary changes such as bronchiectasis, obstructive pneumonia, hemorrhagic infarction etc, background lesions such as anthracosed scar, honeycombing etc, or apparently independent lesions like centrilobular emphysema should be described, and then not occupying but arising site of cancer should be decided on each case, expressing by the bronchial branching number and generation in central cases and the segment number in peripheral ones. PMID- 10824551 TI - [Serum tumor markers for primary lung carcinoma]. AB - A number of serum tumor markers are clinically relevant for primary lung carcinomas. None of them, however, is applied to screening of lung cancers because of their unsatisfactory sensitivity and specificity. Among them, measurements of CEA, CYFRA 21-1, NSE, and pro GRP frequently give subsidiary information as to differential diagnosis, monitoring of treatment, and early detection of recurrence. Current trend of the tumor markers for lung cancer includes development of new markers such as p53 tumor suppressor gene product and I-CTP for detecting bone metastasis. Attempts to detect micrometastases by means of RT-PCR of these marker genes are also discussed. PMID- 10824552 TI - [Molecular biological diagnosis]. AB - Biological behavior of lung cancer was evaluated by basic study. Malignancy Associated Change is the concept that the nuclear features of normal cells in the vicinity of cancer show subtle morphological difference from those of healthy individuals. The difference was recognized by high resolution cytometry and the expression of MAC cells was correlated with the degree of abnormality of chest diseases. Comparative genomic hybridization and fluorescence in situ hybridization were performed to investigate genetic abnormality of. Multiple genetic abnormalities and chromosomal instability showed poor prognosis. Two dimensional electrophoresis was employed to detect the expression of the specific protein of lung cancer. TAO2 was proved to be specific to well differentiated adenocarcinoma. Also, metabolic analysis will be employed for cell analysis. PMID- 10824553 TI - [Video-assisted thoracic surgery(VATS) for lung cancer]. AB - Though the recent prevalence of video-thoracoscopic techniques in thoracic surgery for benign diseases, its application for lung cancer surgery has been still controversial. Diagnostic thoracoscopy for indeterminate pulmonary nodule and exploratory thoracoscopy for the purpose of staging of lung cancer are accepted as useful methods. As for the treatment of lung cancer, complete mediastinal lymph node dissection is difficult by VATS and the advantage of VATS lobectomy has been still unproven. However, pursuit of less invasive approach for lung cancer surgery utilizing new technologies is essential. VATS wedge resection for peripherally tiny cancer may be indicated in near future. PMID- 10824554 TI - [Chemotherapy for lung cancer]. AB - Chemotherapy is currently a primary treatment modality for small-cell lung cancer (SCLC). Combination chemotherapy of anticancer agents improves survival and mortality rather than chemotherapy with single agent. On the other hand, the role of chemotherapy in advanced non-small cell lung cancer(NSCLC) is still controversial. Several meta-analysis demonstrated a small but significant survival benefit of cisplatin-based chemotherapy. In the limited stage SCLC and locally advanced NSCLC, a combination of chemotherapy and thoracic irradiation was found to be superior to chemotherapy alone by several randomized trials and meta-analysis. Concurrent administration of anticancer agents with thoracic irradiation may be optimal treatment. In the last few years, several new agents have demonstrated antitumor activity against lung cancer and randomized trials of these drugs are now under way. PMID- 10824555 TI - [Recent advances of radiation therapy for non-small cell lung cancer]. AB - Radiation therapy(RT) has been frequently used for patients with inoperable or unresectable non-small cell lung cancer(NSCLC). In our series of NSCLC patients without progression of disease at the end of two years after RT, the majority of squamous cell carcinoma patients were alive well after 5 years from RT. On the other hand, half of adenocarcinoma patients had distant failures later. The recent data have indicated that the prognosis for selected favorable patients with unresectable disease will be improved by systemic chemotherapy combined with RT. Furthermore recent innovations in RT have shown promise in the treatment of NSCLC, especially squamous cell carcinoma. For future improvement of the treatment results of NSCLC, it is important that the role of RT be established. PMID- 10824556 TI - [Immunogene therapy for lung cancer]. PMID- 10824557 TI - [Current perspectives of new agents in lung cancer]. AB - Irinotecan(CPT-11), Taxol, Taxotere, vinorelbine and gemcitabine have shown a significant activity in previously untreated non-small cell lung cancer (NSCLC). Cisplatin(CDDP) combined with vinorelbine, gemcitabine or tirapazamine was significantly superior in survival to CDDP alone in the treatment of advanced NSCLC. Patients with NSCLC treated with combination of CDDP and Taxol or vinorelbine lived longer than those treated with conventional CDDP-based chemotherapy. CPT-11, topotecan, taxol and amrubicin have demonstrated to be active against small cell lung cancer(SCLC). Combination of CPT-11 and CDDP have had a higher response rate, and better median survival(13 months) in patients with extensive disease SCLC. Clinical trials of target-based drugs including matrix metalloprotenase inhibitors, anti-angiogenesis, tyrosine kinase inhibitors, farnesyl transferase inhibitors and monoclonal antibodies have been initiated in solid tumors including lung cancer. Development of new anti-cancer agents is essential to improve outcomes of patients with lung cancer. PMID- 10824558 TI - [Strategy of treatment for roentgenographically occult lung cancer]. AB - Roentgenographically occult bronchogenic squamous cell carcinomas(ROCs) are very small hilar type lung cancer that grow superficially and localize mainly in the bronchial wall, and these patients are expected to have an excellent prognosis after resection. When a ROC is limited within bronchoscopic visibility and less than 10 mm in longitudinal extension and its bronchoscopic findings are slight thickening and flat or wavelet, the tumor is a good candidate for photodynamic therapy. When a ROC is limited within bronchoscopic visibility, the tumor is a good candidate for segmentectomy with curative intent. The tumor invading beyond bronchial wall and/or nodal involvement should be treated by standard operation. PMID- 10824559 TI - [Adenocarcinoma]. AB - The treatment of patients with adenocarcinoma of the lung depends on the clinical stage of the disease: surgical therapy in stage I or II, multimodal therapy including surgery in stage IIIA, multimodal therapy including combination of cytotoxic drugs and radiation in inoperable cases. Future therapeutic strategies for lung cancer should include the use of spiral CT, which can detect lung tumors with < 2 cm in diameter, in group health screening for the early diagnosis of adenocarcinoma, and the use of recently new developed anticancer for the treatment of inoperable cases of adenocarcinoma. Compared to the classical combined chemotherapy, future therapeutic modalities should be more effective for improving prognosis and quality of life of lung cancer patients. PMID- 10824560 TI - [Small-cell lung cancer]. AB - Small-cell lung cancer(SCLC) represents 15 to 20% of all lung cancers. Since patients with SCLC usually present with disseminated disease, treatment strategies have focused on systemic therapy. Despite advances in the treatment of SCLC during the 1970s with the use of combination chemotherapy, and in the 1980s with the combination of etoposide and cisplatin plus concurrent radiotherapy, there have been no major advances in therapy in the last decade. There is no proven role for dose escalation, alternating chemotherapy, weekly chemotherapy, or maintenance therapy. The recent advent of new active agents, including irinotecan, provides hope for more effective therapies in the 2000s. This review outlines the current treatment strategies including new agents, thoracic radiotherapy, surgical resection, and prophylactic cranial irradiation. PMID- 10824561 TI - [Large cell carcinoma]. AB - Large cell carcinoma is defined as an undifferentiated carcinoma without the characteristic features of squamous cell, small cell, or adeno-carcinomas. Diagnosis is largely based on exclusion of the other cell types of lung cancer by light microscopy. Large cell carcinoma grows rapidly and is usually quite large by the time it is diagnosed. Most of colony stimulating factor-producing lung cancers previously reported are large cell carcinomas. These colony stimulating factors are considered to contribute to the progressive nature of large cell carcinoma. The radiographic appearance of large cell carcinoma is often larger than 4 cm in diameter. The margins of the mass are poorly defined and lobulated while cavitation remains infrequent. Giant cell carcinoma, which is a variant of large cell carcinoma, is a particularly aggressive subtype with a poor prognosis. Metastases occur fairly late in the course of large cell carcinoma. The sites of metastases (brain, bone, adrenal glands, and liver) are similar to those involved with adenocarcinoma, although the gastrointestinal tract metastases are more commonly involved in large cell carcinoma. PMID- 10824562 TI - [Rare subtype of lung carcinoma]. AB - Rare subtypes of lung cancer, that is, carcinoid, adenoid cystic carcinoma, muco epidermoid carcinoma and adenosquamous carcinoma, were described. These tumors except adenosquamous carcinoma, are generally low malignant and slow growing and arise from upper airway. Complete surgical resection is the best treatment. Sometime tracheobronchoplastic surgery is recommended. Adenosquamous carcinoma is more malignant than adenocarcinoma or squamous carcinoma. PMID- 10824563 TI - [Diagnosis and treatment for metastatic pulmonary tumor]. AB - Metastatic tumors from variable organs are seen in pulmonary tissue because of its unique anatomical situation. In this article, current diagnosis and treatment for metastatic lung tumor are reviewed and discussed including recent topics. Control of metastatic lung tumor by chemotherapy and hormonal treatment has been improved. Furthermore, active applying of Video Assisted Thoracoscopic Surgery induces a change of surgical indication for metastatic lung tumor. Metastatic tumors express oncogene and tumor suppressor gene as well as primary lung cancer. However, the relationship between oncogene/tumor suppressor gene and lung metastatic phenomenon is still unclear. Molecular biological study may indicate new treatment strategy against metastatic pulmonary tumor. Prospective controlled studies for each type of primary tumor are needed to evaluate the true efficacy of pulmonary metastectomy. PMID- 10824564 TI - [Diagnosis and treatment of small peripheral lung cancer]. AB - Recently, routine clinical use of CT scan has enabled the detection of many small pulmonary nodules. Concurrently, high-resolution CT(HRCT) has made great advances in the diagnosis of small peripheral lung cancer. In the tumor of adenocarcinoma, non-invasive bronchioloalveolar carcinoma(BAC) component having a replacement growth pattern of alveolar lining cells shows ground-glass opacity(GGO) on HRCT. On the basis of analysis of small peripheral adenocarcinoma (= < 2 cm), patients with tumors showing > = 50% GGO had no metastatic tendency. Therefore, simple wedge resection may be acceptable as a radical operation. On the other hand, since patients with tumors < 50% GGO had slight metastatic tendency depending upon the tumor size, segmentectomy with mediastinal lymph node sampling should be considered only for tumors = < 15 mm. PMID- 10824565 TI - [Prevention of lung cancer--primary and secondary prevention]. AB - Prevention consists of primary and secondary prevention. For primary prevention of lung cancer, smoking control is considered to be most important and effective. If the present level of smokers in adults Japanese (52.8% in males and 13.4% in females) is decreased to the 50% level of the present level as it was once planned in "the Healthy Japanese 21", 32.3% of lung cancer in males and 7.6% in females (25.5% in both genders) could be prevented. For secondary prevention of lung cancer, periodic lung cancer screening may be useful. If the coverage rate of lung cancer screening is elevated to 30-50%, 15-25% of lung cancer deaths will be prevented assuming the odds ratio of lung cancer death is 0.50 for those who received lung cancer screening. It was estimated that if both smoking control and lung cancer screening were promoted extensively in Japan, as much as 30 to 50% of lung cancer incidences/deaths could be prevented. PMID- 10824566 TI - [Informed consent, QOL and cost benefit in the treatment of lung cancer]. AB - Ethical issues, including informed consent, quality of life and cost/benefit, are discussed. Informed consent is not only ethical discipline but also legal concept. It is important to know how and when to disclose the truth. Training and experience are critically important to acquire clinical-behavioral skill. Physicians also should know some pitfalls, including patient's right to decline information especially in terminal situation. Quality of life(QOL) is an important concept for medical practice in the treatment of lung cancer. Although several instruments have been developed for QOL measurement, poor compliance and feedback to patient care, and meaningless domains are criticized. Disease related symptom(DRS) and cost analysis should be used in future clinical trials of lung cancer. PMID- 10824567 TI - [Epidemiology and control activities of tuberculosis in Japan]. AB - Tuberculosis has declined in Japan for 50 years, however it is more prevalent than other developed countries and speed of decline is much slower recently in Japan than in European countries in 1950-80s when the incidence rate was almost the same as that in Japan now. Most of old cases were reactivation and most of young cases were those soon after new infection. Early case detection and treatment and cutting the chain of transmission is necessary for the reduction of young cases. The defaulter rate among smear positive new cases is 3.6% and is higher among non-skilled workers. DOT has not been widely done on outpatient clinic basis but should be considered for those who cannot stay in hospitals. PMID- 10824568 TI - [Child neurology and rehabilitation]. AB - The history of child neurology and the changing pattern of research methods in this field are reviewed with special reference to holoprosencephaly and recent technical advances in sleep research. This is followed by a discussion on the relationship between child neurology and rehabilitation. The majority of child neurologic disorders are developmental disabilities, but acquired child neurological diseases also show chronic progressive course in many cases. Therefore, child neurologist should understand the basis of rehabilitation approach and appreciate the three classes of disabilities; subsequently, a plan needs to be incorporating medical treatment and a program of rehabilitation for the disabled children. It is important that the role of the various rehabilitation specialists (rehabilitation doctor, physiotherapist, occupational therapist, and others) are understood in relation to the work of pediatric neurologist. Finally, a brief discussion is presented on the rehabilitation approach of patients with hypoxic encephalopathy and the information of welfare equipment. PMID- 10824569 TI - [Developmental disorders and their responsible genes; the genes involved in neuronal positioning]. AB - The mammalian neocortex is a highly ordered structure with different classes of neurons aligning in an organized radial array of six cellular layers ranging from the pial surface to the white matter. Positioning of a particular neuron at the particular cortical layer is an essential step in forming an elaborate neuronal network in the neocortex. It is important to study the molecular mechanism how neuroblasts produced reach to a particular position in the laminated structure of the brain. We took advantage of reeler mutant mouse in which the position of the neurons is abnormal. We obtained an antibody against reeler gene product which we named CR-50, by immunizing wild type embryonic brains to reeler mice. CR50 blocked neuronal positioning in the cerebellum, hippocampus and cerebral cortex. CR-50 reacted with the product of the reelin gene, which was cloned as the responsible gene for the reeler mutant. Cajal-Retzius neurons secrete CR-50 antigen/Reelin and play an important role for neuroblasts to settle to their proper position in the brain. The mutant mouse missing "disabled 1", called yotari mouse, shows similar phenotypes in both morphology and behavior to those of reeler. The information of CR-50 antigen/Reelin is considered to be transmitted inside the neuroblasts to the "disabled 1", which is an adaptor protein to Src, Fyn, Abl tyrosine kinase. PMID- 10824570 TI - [Media for 21st century--towards human communication media]. AB - Today, with the approach of the 21st century, attention is focused on multi-media communications combining computer, visual and audio technologies. This article discusses the communication media target and the technological problems constituting the nucleus of multi-media. The communication media is becoming an environment from which no one can escape. Since the media has such a great power, what is needed now is not to predict the future technologies, but to estimate the future world and take to responsibility for future environments. PMID- 10824571 TI - [Mental retardation: social supports for people with metal retardation from aspects of individual and family life-cycle: introductory remarks] [In Process Citation] AB - In this symposium, the terminology and difinition of mental retardation were discussed. We examined theories and practices of social services, as well as policies for people with mental retardation from the aspects of individual and family life-cycle. Panelists at the invitation of this symposium were experts on this subject. They proposed new theories and made practical comments. The key words include the change of medical to social model; early diagnosis and intervention; medical care for children with severe disabilities at school-life and support for their family; career education and transition services; support with residential facilities and community-based approach; and empowerment of people with developmental disabilities. PMID- 10824572 TI - [Early diagnosis and early intervention for mentally handicapped children]. AB - To support a mentally handicapped child and his/her family, it is very important to understand problems concerning the first-step of life-long help: early detection, intervention and treatment of the child in infancy or toddlers. The author discussed these problems based on his 20-year-long experience in community based approach. PMID- 10824573 TI - [Children with disabilities, families with a disabled child and social policy]. AB - In this article the author described three social movements by families with a disabled child: (1) respite services which refresh family members by a short-term relief care in a local region, (2) after-school activities in which children with disabilities participate in recreation activities and built up a friendship with non-disabled people in neighborhood, and (3) schooling of children with special health care which enables the children to attend unaccompanied by mothers until after-school hours. The author's analysis indicated that a social policy for the people with disabilities and their families should consider other dimensions which may be influenced by the social policy. This is because different social settings (e.g. home, school, family) are interdependent and that events and changes in one unit have an effect on other social settings. PMID- 10824574 TI - [Medical care and support for children with mental retardation in school]. AB - Development of medical knowledge and technology has greatly improved our understanding of mental retardation. However, clinically applicable treatment of mental retardation is still limited. As described in the definition of mental retardation by the American Association of Mental Retardation, functional maladaptations in various aspects of everyday life are the most integral part of disabilities in mental retardation. The role of pediatric neurologist in the medical care for children with mental retardation is not limited in medical treatment. Pediatric neurologists should play an important role as a coordinators in integrated care. Providing of medical care in special schools for children with mental retardation is among the most important issues, since children spend most of their time either at home or in school. The current situation of "medical care" in school, and its problems are discussed. PMID- 10824575 TI - [From school of community: transition services and career education for individuals with developmental disabilities]. AB - This article examines the theory and practice relating to transition services and career education for individuals with developmental disabilities in Japan. The article is also intended to guide future enactment and social services that affect life-cycle transitions. PMID- 10824576 TI - [Support with residential facilities]. AB - In Western countries many residential institutions for the mentally retarded have been built since the end of 19th century. Some large-scaled public facilities accommodated as much as 5,000 to 8,000 residents. This trend persisted the middle of this century. However, institutionalism has been abandoned since the latter half of 1960's. In Japan the law for the welfare of persons with mental retardation was enacted in 1960. Thereafter the most important subject was the construction of residential institutions. Thus, Japan lags behind the United States and European countries by nearly half a century as far as the policy for the mentally retarded is concerned. International Year of Persons with Disabilities in 1981 and subsequent United Nations Decade of Persons with Disabilities provided us with motivation to change the social welfare services in Japan under the concept of normalization. At present Japanese social welfare systems are changing to community-oriented services, such as community residential facilities and day-activity centers. PMID- 10824577 TI - [Empowerment of people with developmental disabilities]. AB - People with developmental disabilities have their own life cycles and social services for them, such as therapeutic training, education, vocational training, and community care, should respect their initiative in decision-making and controlling their own lives. However, social isolation of the disabled people and their families, lack of resources, and discrimination have resulted in their disempowerment. This paper discusses the professionals' role in empowerment, focuses on social and consumer models, and emphasizes the importance of the partnership between professionals and self-help groups. PMID- 10824578 TI - [Nation-wide survey on hemimegalencephaly in Japan]. AB - We report here 38 Japanese patients with hemimegalencephaly collected by a national survey study. All the patients were sporadic. There was no familial occurrence or sex difference. Some patients had basic diseases: hypomelanosis of Ito in 3 cases and organic nevus syndromes in 8. Most patients had hemiparesis, and 11 were bed-ridden. All except for 3 patients had mental retardation, being profound in half of them. There was no correlation between the side of hemimegalencephaly and clinical symptoms. All patients had epileptic seizures, which first appeared within 24 hours after birth in 4 cases, within 7 days in 7, within a month in 2, within 6 months in 10, and within a year in 4. Antiepileptic drugs were not very effective for controlling seizures. In 7 patients, however, functional hemispherectomy resulted in seizure control and improved development. The patients whose epileptic symptom occurred earlier tended to be more severe in clinical symptoms. PMID- 10824579 TI - [Epileptogenesis of acute encephalitis and acute encephalopathy: epilepsy with its onset in the acute phase and without a latent period]. AB - Epileptogenesis was evaluated in 60 patients with acute encephalitis and in 10 patients with acute encephalopathy. Forty-seven patients have been seizure-free during for more than three years' follow-up (Group III). On the other hand, 23 patients developed epilepsy. Among them, 18 patients developed epilepsy after a latent period of 1 month to 2 3/12 years (Group I). In Group I, a younger age of the onset, a long period of disturbed consciousness and a high activity of CSF neuron-specific enolase (NSE) was associated with refractory epilepsy. The other five patients had continuous seizures from the acute phase of encephalitis without a latent period (Group II). They had more than 2 types of partial motor seizures which occurred frequently during the acute phase of encephalitis. The NSE activity in the CSF of patients in Group II was less than 50 ng/ml, being similar to those in Group III. The epilepsy in Group II, however, was the most refractory. The reason for the development of this continuous refractory epilepsy remained obscure. PMID- 10824581 TI - [Brainstem hemorrhage following acute necrotizing encephalopathy]. AB - We report here a case of acute necrotizing encephalopathy (ANE) showing brainstem hemorrhage. A 5-year-11-month-old boy with a history of febrile seizures was admitted to our hospital because of high fever and coma. Laboratory studies on admission demonstrated elevated serum transaminase and blood glucose. Brain CT was normal on admission, but 9 hours later, it showed low density areas in the bilateral thalamus, putamen, midbrain, pons and cerebellum. Brain MRI revealed abnormal short T1 and long T2 signals in the same areas. Despite promptly performed intensive care, massive brainstem hemorrhage occurred on the fourth day. EEG showed a suppression-burst pattern followed by a flat pattern on the sixth day. The patient died on the eighth day of multiple organ failure. This is the first case of massive brainstem hemorrhage that occurred in association with ANE. PMID- 10824580 TI - [Brain lesion in congenital myotonic dystrophy]. AB - Congenital myotonic dystrophy (CMyD) affects the brain, causing mental changes and psychomotor retardation. However, the pathophysiology of the brain dysfunctions in CMyD remain to be clarified. We described two cases of CMyD with brain abnormalities. Case 1 was diagnosed as having ventricular dilation at 17 days after birth, and died at 3 years and 6 months. Case 2 was diagnosed as having ventricular dilation at birth, and died at 1 year and 3 months. Pathologically, both cases showed remote hypoxic ischemic brain damage and leptomeningeal glioneuronal heterotopia (LGH). In our patients, the white matter changes may have been caused by perinatal asphyxia, and LGH by embryological abnormalities. Taken our data and those of previous reports together, it is suggested that cerebral abnormalities in CMyD are ascribed to both hypoxic ischemic changes and histogenetic abnormalities. PMID- 10824582 TI - [Effect of baclofen on intractable hiccup]. PMID- 10824583 TI - [Evaluation of brainstem function with multimodality evoked potentials Joubert syndrome]. PMID- 10824584 TI - Participation of the photosensitizer alpha-terthienyl in the peroxidase-catalyzed oxidation of indole-3-acetic acid. AB - The plant photosensitizer alpha-terthienyl (alpha T) is toxic toward a variety of organisms, and normally requires exposure to ultraviolet-A radiation for activation and singlet molecular oxygen formation. However, some toxicity has also been reported to occur in the dark. One hypothesis that has been proposed to account for this light-independent toxicity is that the sensitizer becomes activated by energy transfer from the excited-state products of enzymatic reactions. We have investigated this hypothesis using the horseradish peroxidase (HRP)-catalyzed oxidation of indole-3-acetic acid (IAA), which generates indole-3 aldehyde in an excited triplet state. Light is emitted during the IAA/HRP reaction at acidic pH, is increased by inclusion of alpha T and is not observed with heat-denatured HRP. The rates of both the oxidation of IAA and the subsidence of light emission are more rapid in the IAA/alpha T/HRP system than with IAA and HRP alone, indicating that the presence of alpha T accelerates the reaction. Bleaching occurs at the wavelength of maximal alpha T absorbance and is promoted by the inclusion of IAA. Readdition of both IAA and alpha T to a spent reaction mixture is required to restore light emission after it has subsided, further suggesting that both are consumed in the reaction. We were unable to detect measurable quantities of singlet molecular oxygen formation in this system. These results do not support the energy transfer hypothesis, but instead are more compatible with a model proposed by Krylov and Chebotareva [Krylov, S. N. and A. B. Chebotareva (1993) FEBS Lett. 324, 6-8] for the co-oxidation of IAA and xanthene dyes. PMID- 10824585 TI - Structural and environmental requirements for quenching of singlet oxygen by cyanine dyes AB - Singlet-oxygen quenching constants were measured for 19 cyanine dyes in acetonitrile. The most efficient quenchers were 1-butyl-2-[2-[3-[(1-butyl-6 chlorobenz-[cd]indol-2(1H)- ylidene)ethylidene]-2-chloro-1-cyclohexen-1 yl]ethenyl]-6-chlorobenz[cd] indolium and 6-chloro-2-[2-[3-(6-chloro-1 ethylbenz[cd]indol-2(1H)-ylidene) ethylidene]-2-phenyl-1-cyclopenten-1 yl]ethenyl]-1-ethyl-benz[cd]indolium, having quenching constants with diffusion controlled values of 2.0 +/- 0.1 x 10(10) and 1.5 +/- 0.1 x 10(10) M-1 s-1, respectively. There was a trend toward increased quenching constants for cyanine dyes with the absorption band maxima at longer wavelengths. However, the quenching constants correlated better with the oxidation potentials of the cyanine dyes, suggesting that quenching proceeds by charge transfer rather than energy transfer. The quenching constants for 1,1',3,3,3',3' hexamethylindotricarbocyanine perchlorate and 1,1'-diethyl-4,4'-carbocyanine iodide were measured in several solvents as well as in aqueous solutions of detergent micelles. In different solvents, the quenching constants varied by as much as a factor of 50. The quenching constants were largest in solvents with the highest values on the pi* scale of Kamlet, Abboud, Abraham and Taft. This was consistent with quenching occurring by charge transfer. Within cells, cyanine dyes concentrate in membrane-bound organelles. The quenching constants were substantial within detergent micelles. To the extent that micelles are models for biological membranes, cyanine dyes may be effective biological singlet-oxygen quenchers. PMID- 10824586 TI - Simultaneous photoconjugation of methylene blue and cis-Rh(phen)2Cl2+ to DNA via a synergistic effect. AB - Irradiation of the red-light absorbing dye, methylene blue (MB), in the presence of the metal complex, cis-Rh(phen)2Cl2+ (BISPHEN), leads to irreversible photobinding of both reagents to DNA. Evidence from absorption and emission spectroscopy indicates that the dye is strongly complexed to the DNA at the concentrations used in the experiments and that this complex is unaffected by the presence of BISPHEN. The level of covalent binding is proportional to the absorbed light dose, with the quantum efficiency for covalent binding of BISPHEN to the DNA with 633 nm light equal to 3.5 x 10(-4). Electrospray ionization mass spectrum of a mixture of DNA fragments created by enzymatic degradation of DNA isolated following irradiation indicates that purine adducts are formed with both BISPHEN and the dye. In addition, UV-Vis and high-performance liquid chromatography analyses of the irradiated MB/BISPHEN/DNA mixture and isolated adducts show extensive conversion of the dye and metal complex to the corresponding N-demethylated and aquated derivatives, respectively. Triplet quenchers for MB, for example oxygen and benzoquinone, inhibit both the photoconjugation and the photochemistry of BISPHEN. A mechanism for the synergistic interaction is proposed that involves photoconjugation of both partners to the DNA following oxidation and reduction via electron transfer between 1MB*/DNA and 3MB*/BISPHEN. PMID- 10824587 TI - Resolved fluorescence of the two tryptophan residues in horse apomyoglobin. AB - The composite fluorescence emission from the two tryptophans (W7 and W14) of horse heart apomyoglobin was explored by fluorescence quenching experiments. The fluorescence of the W7 residue is the only one involved in the quenching by iodide or trichloroethanol (TCE) titration. The fluorescence contribution of W7 is 49% of the total apomyoglobin emission, and its spectrum is red-shifted compared to the W14 emission. The fluorescence decay of Trp residues gives an average fluorescence lifetime of 2.06 ns for W14 and 2.84 ns for W7. The static fluorescence quenching by TCE was used to monitor the individual motions of the two tryptophans in apomyoglobin. The short correlation time of W7 (rho = 3 ns) explains why this residue can experience various environments without having to assume the existence of several protein conformations occurring during its lifetime emission. PMID- 10824588 TI - Excited state properties of alpha,omega-diphenylpolyenes: photophysical and photochemical studies of donor-acceptor diarylbutadienes AB - alpha,omega-Diphenylpolyenes have attracted a great deal of attention as models of retinyl polyenes that are related to natural photoreceptors involved in energy and sensory phototransductions. Of particular interest have been the topics of their excited state electronic structure and spectroscopic properties. However, the exact nature of the lowest excited state in terms of their structure and energetics is not clearly known. Examination of the photophysics and photochemistry of donor-acceptor diphenylpolyenes can aid in understanding the excited states and photoprocesses of linear polyenes. In this paper are described the absorption, fluorescence and photoisomerization studies of donor-acceptor diarylbutadienes, namely: p-(N,N-dimethylamino)-p'-cyano-1,4-diphenylbuta-1E,3E diene (1), p-(N,N-dimethylamino)-p'-nitro-1,4-diphenylbuta-1E,3E-diene (2), p (N,N-dimethylamino)-m'-nitro-1,4-diphenylbuta-1E,3E-diene (3), p-(N,N dimethylamino)-o'-nitro-1,4-diphenylbuta-1E,3E-diene (4). Absorption properties are affected as expected due to mesomeric stabilization by the substituent; however, solvent polarity does not significantly affect the absorption properties of these dienes. In contrast, a pronounced solvatochromic fluorescence behavior of these dienes in organic solvents is observed. Time-resolved fluorescence is characterized by a single exponential fluorescence decay with generally increasing lifetime in polar solvents. The fluorescence quantum yields are very low, particularly in polar solvents, but do not show any clear trend. Irradiation of 1E,3E- 1-4 in organic solvents yields the corresponding 1E,3Z-isomer due to one-photon-one-bond isomerization of the C=C double bond lying closer to the acceptor group. The photoisomerization also depended on the solvent polarity and on the concentration of diene. The photoisomerization efficiency of dienes 1 and 2 under direct irradiation condition is greater than dienes 3 and 4. In comparison to the efficiency of photoisomerization under direct irradiation condition, the photosensitized isomerization efficiency is much less, particularly for dienes 1 and 2. The results are discussed in terms of the involvement of excited-state intramolecular charge transfer and conformationally relaxed polar excited states in the photoprocesses of linear polyenes. PMID- 10824589 TI - The effect of pH on the photophysics and photochemistry of disulphonated aluminum phthalocyanine AB - The results of a study of the effect of pH on the photophysics and photochemistry of di-sulphonated aluminum phthalocyanine (AlPcS2) in aqueous solution are presented. The pH dependence of the triplet quantum yield, fluorescence quantum yield, singlet-oxygen quantum yield, triplet lifetime, fluorescence lifetime and apparent dimerization constants is investigated and the results interpreted in terms of the pH dependence of the nature of the axial ligands. Evidence that the aluminum-axial ligand bond strength, rather than dimer binding energy that determines the extent of dimerization is provided by semi-empirical and ab initio calculations. Possible dimer structures obtained using ab initio calculations are discussed. PMID- 10824590 TI - The influence of water on the photophysical and photochemical properties of Piroxicam in AOT/iso-octane/water reversed micelles AB - The photophysical properties of Piroxicam, a nonsteroidal anti-inflammatory drug (NSAID), were investigated at different pHext values in reversed micelles of Aerosol-OT (AOT) in iso-octane, using both steady-state and picosecond time resolved fluorescence spectroscopy. In contrast with the very complex data obtained in aqueous media, where several prototropic species are in equilibrium, the reversed micellar system essentially favors two species. The absorption spectra shows only one isosbestic point at lambda = 348 nm. Excited-state intramolecular proton transfer (ESIPT), also detected in water, is promoted at low water pool contents measured by omega 0 = [H2O]/[AOT]. A strongly shifted (lambda em = 470 nm) tautomeric emission is found. Upon the gradual increase of omega 0, striking differences with pHext are found. At pHext = 4, the drug preferentially locates itself in the interfacial region partitioning between a hydrophobic and a hydrophilic domain. Global analysis was applied to the decay data and the results were interpreted by the "two-state excited-state" formalism. At pHext = 7, the anionic species is prevalent and the probe locates itself deeper inside the water core of the reversed micelles. Thus, a strong dependence on water content is detected, approaching a behavior similar to that observed in free aqueous solutions. PMID- 10824591 TI - Cyanine dyes as protectors of K562 cells from photosensitized cell damage. AB - Several cyanine dyes were found to protect K562 leukemia cells against toxicity mediated by cis-di(4-sulfonatophenyl)diphenylporphine (TPPS2) and light. Most cyanine dyes derived from dimethylindole were better photoprotectors than cyanine dyes with other structures. This correlated with the fact that cyanine dyes derived from dimethylindole were predominately monomeric at millimolar concentrations within K562 cells, while other cyanine dyes formed aggregates. For cyanine dyes that are derived from dimethylindole and have absorption band wavelengths greater than 700 nm, fluorescence-energy transfer from TPPS2 to the cyanine dye was the most important mechanism for photoprotection. There was no spectroscopic evidence for complex formation between the cyanine dyes and TPPS2. The dimethylindole derivative, 1,1',3,3,3',3'-hexamethylindodicarbocyanine, was an excellent photoprotector, but a poor quencher of TPPS2 fluorescence and a relatively poor singlet-oxygen quencher. This cyanine dye may act by quenching excited triplet TPPS2. Singlet-oxygen quenching may contribute to the photoprotection provided by cyanine dyes not derived from dimethylindole. Differences in the subcellular distribution of the various cyanine dyes studied may have contributed to the different apparent mechanisms of photoprotection. PMID- 10824592 TI - The use of silver-stained "comets" to visualize DNA damage and repair in normal and Xeroderma pigmentosum fibroblasts after exposure to simulated solar radiation. AB - The alkaline and neutral comet assays have been widely used to assess DNA damage and repair in individual cells after in vivo or in vitro exposure to chemical or physical genotoxins. Cells processed under neutral conditions generate comets primarily from DNA double strand breaks, whereas under alkaline conditions, comets arise from DNA single and double strand breaks and alkali-labile lesions. A modified version of the alkaline comet assay, as described here, used silver stain to visualize the comets and a Gelbond base to facilitate the manipulation and processing of samples. To demonstrate how these modifications improve the assay, fibroblasts derived from both normal and Xeroderma pigmentosum (Xp) individuals were exposed to simulated solar radiation and the resulting DNA damage and repair evaluated and compared with results from the relevant literature. Comets from normal fibroblasts reached their maximum length at about an hour after irradiation. Dose-dependent increases in comet length were observed up to at least 360 mJ/cm2. In contrast, comet lengths from repair deficient Xp fibroblasts were shorter than normal cells reflecting their reduced capacity to generate single strand breaks by the excision of DNA dimers. For incubation times of more than 1 h, comet lengths from normal fibroblasts underwent a time dependent decrease, supporting the contention that this change was related to the ligation step in the DNA repair process. These changes were compatible with the model of DNA damage and repair established by others for ultraviolet radiation. PMID- 10824593 TI - A sunscreen-tanning compromise: 3D visualization of the actions of titanium dioxide particles and dihydroxyacetone on human epiderm. AB - The self-tanning agent dihydroxyacetone (DHA) was applied to human skin samples, and its effect on light absorption was followed in time to study the DHA influence inside the different layers of skin. Application of DHA shows increased light absorption in the visible light region, as evidenced by skin tanning. The tanning effect is enhanced by UV irradiation and appears localized in the near stratum corneum layer as revealed by depth analysis of the time signal. As a reference, application of an emulsion containing titanium dioxide particles shows clear surface stability and a screening of light penetration beyond the stratum corneum. PMID- 10824594 TI - Development and intercalibration of ultraviolet solar actinometers AB - Ultraviolet (UV) sunlight actinometers were developed based on the photolysis of nitrate and nitrite. Photon exposures (i.e. time-integrated irradiances) were quantified from the photochemical production of salicylic acid (SA) or p hydroxybenzoic acid (pHBA) formed by the reaction of the hydroxyl radical with benzoic acid. The solar response bandwidth for the nitrate actinometer in quartz tubing was 322 +/- 11 nm during the Spring of 1999, while the response bandwidth of the Mylar D-filtered nitrite actinometer was 355 +/- 25 nm. Intercomparisons of the nitrate and nitrite actinometers with a Smithsonian Environmental Research Center SR-18 scanning UV-B radiometer (SERC SR-18) and an Optronics OL-754 spectroradiometer (OL-754) were performed during the summer of 1998, and the winter and spring of 1999. Photon exposures determined by the nitrate actinometer were in excellent agreement with the SERC SR-18, with a slope (95% confidence interval [CI]) of 0.98 +/- 0.01 based on SA production and 0.94 +/- 0.02 based on pHBA production. Excellent agreement was also found between the nitrite actinometer and the OL-754, with a slope (95% CI) of 1.00 +/- 0.01 using SA production and 1.00 +/- 0.02 using pHBA production. These actinometers are well suited for use in the water column and are sufficiently sensitive to determine photon exposures below the 0.1% UV lightlevel. PMID- 10824595 TI - Oxygen uptake photosensitized by disorganized chlorophyll in model systems and thylakoids of greening barley. AB - Light-dependent oxygen uptake was observed and studied in thylakoids from early greening barley in comparison to oxygen uptake in chlorophyll solutions and in thylakoids from fully green leaves. Substantial oxygen uptake was observed in chlorophyll solutions supplemented with tryptophan, histidine, ascorbic acid or linoleic acid. This uptake was diminished by adding azide, beta-carotene and alpha-tocopherol, which are specific singlet-oxygen quenchers. Illuminated thylakoids from greening barley also exhibited marked oxygen uptake that, likewise, was strongly quenched by azide. In comparison, azide was found not to affect oxygen uptake that is associated with the methyl viologen-catalyzed Mehler reaction. It is reasoned that in the first two cases the oxygen uptake arises from chlorophyll-photosensitized activation of oxygen to the singlet state and its consumption by exogenous or endogenous substrates. In greening, we propose that disorganized chlorophyll photo-sensitizes the oxygen uptake. PMID- 10824596 TI - In vitro fluorescence, toxicity and phototoxicity induced by delta-aminolevulinic acid (ALA) or ALA-esters. AB - Synthesis of delta-aminolevulinic acid (ALA) derivatives is a promising way to improve the therapeutic properties of ALA, particularly cell uptake or homogeneity of protoporphyrin IX (PpIX) synthesis. The fluorescence emission kinetics and phototoxic properties of ALA-n-pentyl ester (E1) and R,S-ALA-2 (hydroxymethyl) tetrahydrofuranyl ester (E2) were compared with those of ALA and assessed on C6 glioma cells. ALA (100 micrograms/mL), E1 and E2 (10 micrograms/mL) induced similar PpIX-fluorescence kinetics (maximum between 5 and 7 h incubation), fluorescence being limited to the cytoplasm. The 50% lethal dose occurred after 6 h with 45, 4 and 8 micrograms/mL of ALA, E1 and E2, respectively. ALA, E1 and E2 induced no dark toxicity when drugs were removed after 5 min of incubation. However, light (25 J/cm2) applied 6 h after 5 min incubation with 168 micrograms/mL of each compound induced 85% survival with ALA, 27% with E1 and 41% with E2. Increasing the incubation time with ALA, E1 and E2 before washing increased the phototoxicity, but E1 and E2 remained more efficient than ALA, regardless of incubation time. ALA-esters were more efficient than ALA in inducing phototoxicity after short incubation times, probably through an increase of the amount of PpIX synthesized by C6 cells. PMID- 10824597 TI - Role of the p38 and MEK-1/2/p42/44 MAP kinase pathways in the differential activation of human immunodeficiency virus gene expression by ultraviolet and ionizing radiation. AB - Ultraviolet (UV) radiation is a potent activator of human immunodeficiency virus (HIV) gene expression in a HeLa cell clone having stably integrated copies of an HIV cat (cat gene under control of the HIV promoter) reporter construct, whereas ionizing radiation is ineffective. UV-activated HIV gene expression is completely blocked by the specific p38 mitogen-activated protein (MAP) kinase inhibitor SB203580 and by expression of a kinase-inactive p38 mutant that interferes with normal p38 function, suggesting that this stress-activated protein kinase plays an important role in UV-mediated transcriptional activation of HIV. In support of these findings, we show here that Western blot analysis demonstrated rapid and significant activation of p38 MAP kinase by UV. On the other hand, gamma radiation activated p38 MAP kinase very poorly in HeLa cells at both low and high doses at times (5-30 min) when UV radiation was effective. UV radiation also activated HIV gene expression (< or = 9-fold) in 1G5 Jurkat T-cells stably transfected with a luciferase reporter gene under control of the HIV promoter. In these cells, gamma-radiation stimulated HIV gene expression but to a lesser extent (< or = 3-fold) and with different kinetics than after UV radiation, and this response was obliterated by the incubation of cells with the mitogen activated protein kinase/Erk kinase (MEK)-1/2 inhibitor PD98059. This result suggests that in these cells signaling in response to gamma-radiation is transduced through the MEK-1/2/p42/44 MAP kinase pathway to increase HIV gene expression. All combined, these results suggest that activation of p38 MAP kinase is necessary for efficient HIV gene expression triggered by DNA damaging agents, and, in a cell type-specific manner, activation of the MEK-1/2/p42/44 MAP kinase pathway is important for triggering a response to gamma-radiation. Thus, it appears as if UV signaling leading to HIV gene expression requires the p38 MAP kinase pathway whereas activation by gamma-radiation requires the MEK-1/2/p42/44 MAP kinase pathway. PMID- 10824598 TI - Plasma membrane associated location of sulfonated meso-tetraphenylporphyrins of different hydrophilicity probed by total internal reflection fluorescence spectroscopy. AB - Sulfonated meso-tetraphenylporphyrins of different hydrophilicity were microspectrofluorimetrically examined in endothelial cells using total internal reflection (TIR) illumination or epi-illumination. Since the penetration depth of the evanescent field during TIR illumination is limited to a few hundred nanometers, photosensitizers were almost selectively examined in close vicinity to the plasma membrane. Pronounced fluorescence signals during TIR illumination were observed for the hydrophilic compounds meso-tetraphenylporphyrin tetrasulfonate (TPPS4) and meso-tetraphenylporphyrin trisulfonate (TPPS3), whereas the more lipophilic compounds meso-tetraphenylporphyrin disulfonate (TPPS2a) and meso-tetraphenylporphyrin monosulfonate (TPPS1) could only be detected under epi-illumination. Irradiation of TPPS1 and TPPS2a in the Soret band led to an increase in fluorescence intensity and formation of a photoproduct with an emission maximum around 610 nm, which was limited to intracellular compartments. In contrast, fluorescence spectra of TPPS3 and TPPS4 obtained by TIR and epi-illumination remained almost unchanged after irradiation in the Soret band. Extralysosomal location of TPPS3 and TPPS4 in close proximity to the plasma membrane was deduced from experiments with the lysosomal markers acridine orange (AO) or lysotracker yellow (LY), which were not detectable under TIR illumination. In conclusion, these results provide for the first time direct evidence for a plasma membrane-associated fraction of the hydrophilic compounds TPPS3 and TPPS4 in living cells. PMID- 10824599 TI - Relationship between vitamin D status and skin phototype in general adult population. AB - A link between bone mineral density and skin color has been reported recently, and pigmentation has been shown to affect cutaneous vitamin D production. In the present study, we investigated the relationship between phototype, global self assessed sun exposure, geographical location and vitamin D serum levels in 1191 French adults. When the factors were analyzed separately, individuals with lower phototypes as well as those with lower sun exposure showed significantly lower levels of vitamin D than those with darker phototypes or those with higher sun exposure. However, when factors were analyzed as a whole, the vitamin D status was no longer linked with the phototype, but with sun exposure and geographical location. Since phototypes and global self-assessments of sun exposure were positively linked, our data suggest that lower vitamin D levels in fair-skinned individuals are due to their sun exposure behavior. PMID- 10824600 TI - Heat and concentration effects on the small heat shock protein, alpha-crystallin. AB - alpha-Crystallin, a major protein of the mammalian lens, plays a vital role in maintaining the structural stability and transparency of the lens. It performs this function through chaperone-like activity; it has recently been reported that heating alpha-crystallin enhances this ability. The present studies, using both time-resolved and steady-state fluorescence methods, were carried out to compare the conformational changes that result from heating with those that result from increasing protein concentration (up to 70 mg/mL). The relative fluorescence quantum yield from tryptophan (Trp) present in alpha-crystallin increases and then decreases with a concomitant shift of the emission maximum to longer wavelengths when either heating times or protein concentrations are increased. The time profile of fluorescence decay was resolved into three components with lifetimes of ca 0.5, 3 and 7 ns and emission maxima of ca 340, 342 and 350 nm, respectively. With longer heating time or increasing concentrations the contribution from the longer-lived component increases at the expense of the shorter-lived species. These data indicate that with heating or at higher concentrations the internal Trp residues move to the surface of the protein giving a more hydrophobic exterior and possibly explain the reported increased chaperone activity upon heating. As a result of the concentration studies, alpha crystallin may be more efficient in its chaperone activity in vivo than has been determined by in vitro experiments. PMID- 10824601 TI - Photocurrent from oriented membrane films containing acid-blue and acid-purple bacteriorhodopsin and its mutants. AB - This paper investigates the fast photocurrent components, B1 and B2, from oriented bacteriorhodopsin (BR) membrane films at low pH, under pulsed laser excitation. Adding chloride ion changes the acid-blue BR to its acid-purple form. In the presence of chloride, the acid-purple BR shows a positive B2 component in the same direction as that of BR at neutral pH, indicating a rapid intramolecular charge transfer. In the absence of chloride, the acid-blue BR shows only a negative B1 with multi-components, indicating a rapid charge separation process associated with retinal photoisomerization. The multi-components in B1 are possibly formed due to the heterogeneity of the acid-blue BR. In addition, BR mutants, D85N and D115N, at low pH and in the presence of chloride, generate the B2 component as well. The observation of chloride-dependent B2 component in various cases at low pH, is in favor of a possible transient chloride ion transfer, although the nature of the charge being transferred cannot be identified so far. PMID- 10824602 TI - Two photobiological pathways of phytochrome A activity, only one of which shows dominant negative suppression by phytochrome B. AB - The plant receptor phytochrome A (phyA) mediates responses like hypocotyl growth inhibition and cotyledon unfolding that require continuous far-red (FR) light for maximum expression (high-irradiance responses, HIR), and responses like seed germination that can be induced by a single pulse of FR (very-low-fluence responses, VLFR). It is not known whether this duality results from either phyA interaction with different end-point processes or from the intrinsic properties of phyA activity. Etiolated seedlings of Arabidopsis thaliana were exposed to pulses of FR (3 min) separated by dark intervals of different duration. Hypocotyl growth inhibition and cotyledon unfolding showed two phases. The first phase (VLFR) between 0.17 and 0.5 pulses.h-1, a plateau between 0.5 and 2 pulses.h-1 and a second phase (HIR) at higher frequencies. Reciprocity between fluence rate and duration of FR was observed within phases, not between phases. The fluence rate for half the maximum effect was 0.1 and 3 mumol.m-2.s-1 for hourly pulses of FR (VLFR) and continuous FR (HIR), respectively. Overexpression of phytochrome B caused dominant negative suppression under continuous but not under hourly FR. We conclude that phyA is intrinsically able to initiate two discrete photoresponses even when a single end-point process is considered. PMID- 10824603 TI - Changes in hemolymph glutathione status after variation in photoperiod and light irradiance in crayfish Procambarus clarkii and Procambarus digueti. AB - This work studied the effect of light-stressors, irradiance and photoperiod length on the status of hemolymph glutathione in two species of crayfish, Procambarus clarkii and Procambarus digueti. Adult animals of each species were submitted to two experimental approaches: (1) two batches of each species were placed under low or high light irradiant conditions of light-dark (LD) 24 h cycles of two different photoperiod lengths, one normal LD 12: 12 and one extreme LD 20:4 low and high irradiance for 10 weeks. Time-dependent light changes on hemolymph glutathione concentration were determined throughout the entire experimental period; and (2) three batches of the two species were submitted to independent treatments consisting of the same LD 12:12 cycles of high and low irradiance and 20:4 high-irradiance LD cycles. Reduced and oxidized glutathione hemolymph concentrations were determined and total glutathione was calculated. In addition midgut glutathione reductase activity in both species was determined. The two species showed different hemolymph glutathione reactivity and glutathione status for the two light parameters. Dissimilar responses of both species, as well as the rate of mortality of P. digueti represent specific differences in the metabolic responses, as well as tolerance to photo-oxidative stress produced by light. The role of glutathione in the tolerance of crayfish to photo-oxidative stress is discussed. PMID- 10824604 TI - A novel prokaryotic UVB photoreceptor in the cyanobacterium Chlorogloeopsis PCC 6912. AB - We present evidence for the presence and nature of a UVB-specific photoreceptor in the cyanobacterium Chlorogloeopsis PCC 6912. The photoreceptor mediates at least the photosensory induction of mycosporine-like amino acid (MAA) synthesis. Because MAA synthesis in this organism can also be induced under salt stress, we could distinguish between the photosensory and the purely biochemical requirements of MAA synthesis. Neither visible light nor UV radiation was necessary for the biosynthetic process, thus indicating that the UVB (280-320 nm) dependence of biosynthesis is based on a UV photosensory capacity of the organism. An action spectrum of the MAA synthesis showed a distinct peak at 310 nm tailing down into the UVA (320-400 nm) region with no detected activity above 340 nm. We found that radiation below 300 nm caused significant inhibition of synthesis of MAAs indicating that the action spectrum at these wavelengths may not have been satisfactorily resolved. We propose that a pterin is a good candidate for a photoreceptor chromophore as (1) reduced pterins present absorption spectra congruent with the action spectrum obtained; and (2) an inhibitor of the biosynthetic pathway of pterins and an antagonist of excited states of pterins, both depressed the photosensory efficiency of induction but not its chemosensory efficiency. PMID- 10824605 TI - Biological basis for anti-angiogenic therapy, 8-10 November 1999, Milan, Italy. PMID- 10824606 TI - Assessment of different methods for staining Helicobacter pylori in endoscopic gastric biopsies. AB - The recent implication of Helicobacter pylori in the pathogenesis of gastritis peptic ulcer syndrome and its relevance for the development of upper gastrointestinal malignancy warrant efficient methods for the detection and demonstration of the organism in biopsy specimens. We have compared 5 staining methods, namely, haematoxylin and eosin (H & E), immunohistochemistry (IHC), the silver staining HpSS, the alcian yellow-toluidine blue (Leung) method (A-Y) and Genta staining, for the demonstration of the organism in gastric biopsies taken from antrum, body and fundus of 118 patients who presented to our hospital with upper gastrointestinal symptoms. We found no significant differences in the efficacy of H & E, IHC, HpSS and A-Y in the demonstration of H. pylori in all 3 gastric sites. The least reproducible stain in our hands was the Genta stain. We conclude that H & E is adequate for the initial assessment of gastric biopsies in symptomatic upper gastrointestinal patients. This is because it is a well-tested, cheap and easy staining method, requiring a relatively short period of time to perform, with highly reproducible results. It has an added advantage of enabling simultaneous assessment of morphological changes accompanying H. pylori infection. When the density of the organism is expected to be low, we recommend addition of HpSS staining because of its high sensitivity and low cost. The disadvantages of the other staining methods (IHC, A-Y and Genta) are discussed. PMID- 10824607 TI - Immunogold labelling of PCNA and Ki-67 antigen at the ultrastructural level in laryngeal squamous cell carcinoma and its correlation with lymph node metastasis and histological grade. AB - Streptavidin-gold was used for the immunolocalization of PCNA and Ki-67 antigen at the ultrastructural level with a postembedding technique in biopsies of 15 patients with laryngeal squamous cell carcinoma. Positive immunoelectron staining was obtained in 9 cases for PCNA (60%) and in 8 cases for Ki-67 (53%). PCNA was predominantly found in heterochromatin of the nucleus of laryngeal carcinoma cells in a granular pattern. Positivity for PCNA was not found in nucleoli. In 4 cases, positive staining was observed both in nucleus and cytoplasm. In the cytoplasm, it was found to be present on the endoplasmic reticulum and on ribosomes throughout the cytoplasm. Ki-67 antigen was localized in the nucleus where it was associated with heterochromatin and euchromatin. It was also observed in nucleoli in all cases. Cytoplasmic localization of Ki-67 antigen was similar to that of PCNA. All 8 cases that were positive for Ki-67 were also positive for PCNA. Control incubations did not result in labelling with steptavidin-gold particles for both antigens. A significant correlation between PCNA and Ki-67 expression in association with pathological characteristics such as nodal status and histological grade was not found. Our data indicate that Ki 67 antigen staining correlates with PCNA labelling, whereas a relationship between proliferation markers and tumour progression was not found. PMID- 10824608 TI - Anti-rat CD45RA monoclonal antibodies cross-react with glucagon. AB - Anti-rat CD45RA monoclonal antibody (MAb) has been used as a marker for rat B lymphocytes (B-cells). When we applied the MAb to identify B-lymphocytes among immune cells infiltrating pancreatic islets of the BB rat, which is an animal model for human type 1 diabetes mellitus, a subpopulation of islet cells was immunostained as well. Our immunohistochemical investigations demonstrated that the stained islet cells are pancreatic alpha-cells and the MAb cross-reacts with glucagon. PMID- 10824609 TI - Immunohistochemical localization of calcium-binding proteins (CaBPs) in the epidermis of the earthworm Lumbricus terrestris (Annelida, Oligochaeta). AB - The present immunohistochemical study provides the first evidence of the presence of calcium-binding proteins (CaBPs) in the epidermis of the earthworm Lumbricus terrestris (Annelida, Oligochaeta) a lower invertebrate. The entire epidermis was labelled for calmodulin which is in agreement with its ubiquitous occurrence. Immunopositivity for calbindin D28K was limited to mucous cells, while that for S 100 protein was present only in neuroendocrine-like small granular cells. Finally, labelling for parvalbumin was specifically present in the subcutaneous nerve plexus. S-100 protein is considered to be a marker of neuroendocrine cells, at least in lower invertebrates such as Annelida. Although calbindin D28K is considered to be a marker of these cells in vertebrates, the same function cannot be attributed in Lumbricus terrestris. However, we can conclude that S-100 protein, as a regulatory protein, is phylogenetically older than calbindin D28K. We assume that the latter has an autoregulatory function in secretory processes. In agreement with previous data, we suggest that small granular cells exert a paracrine action in osmoregulatory and secretory processes. PMID- 10824610 TI - Localization of cytochrome P450 aromatase and estrogen receptors alpha and beta in testicular cells--an immunohistochemical study of the bank vole. AB - Age- and light-dark cycle-induced changes in immunoexpression of aromatase and estrogen receptors alpha and beta were studied in testes of a seasonally breeding rodent, the bank vole. Seasonal breeding can be mimicked by exposure to different light cycle regimes. In testes of animals that were exposed to long light cycles of 18 h light and 6 h darkness aromatase was in Leydig cells and seminiferous tubules, mainly in spermatocytes, whereas in animals exposed to short light cycles (6 h light and 18 h darkness), only Leydig cells exhibited positive immunostaining for aromatase. Whatever the age of animals, immunostaining for estrogen receptor alpha was restricted to Leydig cells, whereas estrogen receptor beta immunoreactivity was mainly confined to Sertoli cells of both of immature and adult animals, independently of the regimes of light. Additionally, in testes of animals that were exposed to long light cycles, estrogen receptor beta immunoreactivity was observed in seminiferous tubules. Nuclei of germ cells, predominantly spermatocytes and elongated spermatids, were strongly positive which correlated well with aromatase immunoreactivity. As far as we know, the present study is the first study that describes immunoexpression of aromatase and both estrogen receptors alpha and beta in testis of the bank vole. We provide strong evidence that estrogens are not only produced in Leydig cells but also in germ cells in this rodent. These female hormones may play a physiological role in testis, likely in the development of germ cells during spermatogenesis. PMID- 10824611 TI - Localization of the progesterone receptor in the porcine ovary. AB - Regulation of progesterone receptor (PR) expression has been studied in many species. However, precise studies have not yet been performed in the porcine ovary. We have examined the localization of PR in follicles and corpora lutea of the porcine ovary at different stages of their development. The effects of LH and FSH on PR expression in granulosa cells of small antral follicles was also studied. Immunohistochemistry was applied to determine the distribution of PR while immunoblot analysis showed that two isoforms A and B were present. Early antral follicles contained PR in the granulosa layer. In granulosa cells of small and medium antral follicles PR was not detected whereas it was present in the theca layer. Before ovulation, PR was found in both granulosa and theca cells of large follicles and the staining intensity was very strong. FSH or LH treatment of small follicles (100 ng/ml) induced changes in cellular distribution patterns of PR. In both cases, PR was expressed in granulosa cells. PR was detected in corpora lutea in all 3 stages of the luteal phase. Our data show that in the pig ovary changes in PR localization are stage-specific and suggest that expression of PR is positively regulated by both LH and FSH. PMID- 10824612 TI - Lectin histochemical detection of sulfoglycans in the zona pellucida of mammalian antral oocytes. AB - Sulphated esters are important to increase effectiveness of specific biological activities of carbohydrates. Biochemical studies revealed the presence of distinct sulphated glycoproteins in mammal zona pellucida (ZP) that bind proacrosin and thus participate in the sperm-egg fusion processes. In the present study, 6 lectin-horseradish peroxidase conjugates (SBA, PNA, RCA-I, GSA-IB4, GSA II and DBA) were used in combination with desulphation and sialidase digestion to identify sulphocarbohydrates in the terminal and/or subterminal position of oligosaccharide side chains of glycoproteins in the ZP of bovine, ovine, caprine and porcine antral oocytes. In particular, we identified the following terminal sulphoglycans located in the outer layer of the ZP only: SO4-GalNAc in bovine ZP; SO4-Galbeta1,3GalNAc in bovine and ovine ZP; SO4-Galbeta1,4GlcNAc in bovine, ovine and caprine ZP; SO4-alpha-Gal in bovine, caprine and porcine ZP. Subterminal sulphoglycans linked to sialic acid residues were evenly distributed throughout the entire thickness of the ZP: Neu5Ac-SO4-Galbeta1,3GalNAc in bovine and porcine ZP; Neu5Ac-SO4-Galbeta1,4GlcNAc in caprine ZP; Neu5Ac-SO4-alpha-Gal in porcine ZP; Neu5AcSO4-GlcNAc in bovine ZP. The results demonstrate that the chemical composition of the ZP differs among species determining the species specificity of gamete interactions. PMID- 10824613 TI - Neuronal expression of Raf protooncogene in the brain stem of adult guinea pig. AB - The Raf protooncogenes encode for cytoplasmic serine/threonine-specific protein kinases which can be activated via growth factor receptors by phosphorylation. Immunohistochemical and Western blotting studies have proven the existence of Raf protein kinases in neurons of the cerebral cortex of rats and guinea pigs. The aim of the present study was to map the immunohistochemical distribution of Raf kinase-like staining in the brain stem of guinea pig. Polyclonal antibodies were used that were raised against a recombinant viral protein in combination with the avidin-biotin-peroxidase system for detection of immunoreactivity. Specificity of the antibodies was tested in Western blotting experiments. Cytoplasmic immunostaining was observed in motor nuclei of hypoglossal, accessory, vagus, facial, trigeminal, abducent, oculomotor and trochlear nerves, and in the nucleus ambiguus, nucleus retroambigualis, lateral vestibular nucleus, mesencephalic nucleus of the trigeminal nerve, the red nucleus, raphe nuclei and reticular formation. Scattered neurons were stained in other sensory nuclei, such as solitary tract nuclei, medial, dorsal and ventral vestibular nuclei and cochlear nuclei. The spinal trigeminal nucleus and the main sensory nucleus of the trigeminal nerve contained few medium-sized immunoreactive cells. In general, staining was mainly somatodendritic; the axonal plexus was not positive. It is concluded, that the widespread neuronal appearance of cytoplasmic Raf kinase suggests an important role in transmission of trophic and growth factor signals in these neurons. PMID- 10824614 TI - Lectin histochemistry and ultrastructure of microgranular cells in Cinachyra tarentina (Porifera, Demospongiae). AB - A histochemical study is described that characterizes microgranular cells of the demosponge Cinachyra tarentina (C. tarentina) with the use of routine staining methods for mucosubstances, lectin histochemistry and electron microscopy. Microgranular cells are rare or absent in other species of sponges, but abundant in this species. Microgranular cells are present in both ectosome and mesohyl, particularly along the canal of the aquiferous system and around spicule holes. Inclusions of microgranular cells and the extracellular matrix were particularly positive for acidic glycoproteins with abundant sulfated ester groups and glycosidic residues containing GalNAc and Galbeta1,3GalNAc. Terminal L-fucose bound to the penultimate GalNAc residues and/or difucosylated oligosaccharides were present as well. Our results suggest that soybean lectin (SBA), peanut lectin (PNA), and winged pea lectin (WPA) are valuable markers for identifying microgranular cells of C. tarentina. Electron microscopy revealed some of the microgranular cells to contain small smooth cytoplasmic vesicles originating from the Golgi complex and few electron-dense granules, others were characterized by numerous secretory granules and vacuoles formed by vesicle fusion and connected with the plasma membrane. Our results suggest that microgranular cells in C. tarentina contribute to the synthesis of glycoprotein components of the extracellular matrix. PMID- 10824615 TI - Changes in succinate dehydrogenase (SDH) activity in Paramecium caudatum cells in culture as function of age. AB - SDH activity and its frequency distributions were determined cytophotometrically to investigate the change in respiratory oxidative energy metabolism in Paramecium caudatum (P. caudatum) cell populations during their growth in cell culture. Cells from 3 separate cultures were examined on day 2, 5 and 11 post inoculation as measures of logarithmic, early and late stationary phases, respectively. SDH activity per individual cell is expressed as the increase in total absorbance (TA) at 590 nm of nitroblue tetrazolium (Nitro BT) formazans per min per cell area in a specimen (deltaTA/min/microm2) to exclude the influence of cell size on the data. On day 5, the mean SDH activity was higher, being approx 140% of that on day 2 and decreased significantly to only approx 11% at day 11. As the mean SDH activity rose, a certain portion of the cells demonstrated a wider range of activity than on day 2 leading to an increase in the width of the SDH activity-frequency distribution. Moreover, on day 11, approx 85% of cells shifted toward the lowest range of activity with further increase in width of the distribution as the mean activity declined. These findings suggest that the respiratory oxidative energy metabolism in P. caudatum cells rise while the cells change from logarithmic to early stationary phases and decays during late stationary phase with increase in its extent of variety within cell populations. PMID- 10824616 TI - Pediatric nuclear medicine, Part II: Common procedures and considerations. AB - OBJECTIVE: This paper introduces technologists to pediatric nuclear medicine applications as well as serves as a review of the principles of pediatric imaging for more experienced technologists. After reading this article the nuclear medicine technologist should be able to: (a) identify pediatric populations commonly evaluated with nuclear medicine procedures; (b) state the indications for performing pediatric nuclear medicine procedures; and (c) discuss strategies and tips for performing nuclear medicine procedures on pediatric patients. PMID- 10824617 TI - Nuclear pharmacy, Part II: Nuclear pharmacy practice today. AB - OBJECTIVE: Nuclear pharmacy is a specialty within the profession of pharmacy that focuses on the proper use of radiopharmaceuticals. This article reviews various features of contemporary nuclear pharmacy practice. After reading this article the nuclear medicine technologist should be able to: (a) describe nuclear pharmacy training and certification; (b) discuss nuclear pharmacy practice settings; (c) discuss nuclear pharmacy practice activities; (d) list professional organizations; and (e) describe activities associated with job satisfaction. In addition, the reader should be able to discuss regulatory issues of current concern. PMID- 10824618 TI - Evaluation of ocular tumors with technetium-99m-MIBI: planar pinhole technique or SPECT? AB - OBJECTIVE: This study compares 2 imaging protocols, planar pinhole technique (PPHT) and SPECT, for evaluating ocular masses with 99mTc-MIBI. METHODS: Sixteen patients with ocular lesions were studied. Planar images were acquired 10 min after the injection of 740 MBq 99mTc-MIBI with an LFOV camera fitted with a pinhole collimator (5.0 mm). A SPECT study was performed immediately after the planar study, using a 360 degrees orbit, 64 steps, 20 s/stop, a 128 x 128 matrix, and a low-energy high-resolution (LEHR) collimator. Twelve lesions (9.5-18.0 mm) proved to be malignant: 8 primary tumors (ocular melanoma); 3 local relapses of different tumors of the conjunctiva; and 1 ocular metastasis from breast cancer. The remaining 4 lesions (10.0-16.0 mm) were benign: 1 inflammatory lesion; 1 benign intraocular calcification; and 2 naevi. RESULTS: SPECT images showed 11 of 12 malignant lesions (91.6%), whereas the planar technique demonstrated only 4 of the 12 lesions (33.3%). One false-positive result, the inflammatory lesion, was visualized by both techniques. The remaining benign lesions were not detected with either method. CONCLUSION: Technetium-99m-MIBI SPECT is a sensitive technique for detecting malignant ocular tumors. SPECT imaging is a better alternative to planar imaging for ocular tumors. PMID- 10824619 TI - The influence of flood source placement on radiation exposure during quality control testing. AB - OBJECTIVE: This study examined the photon energy distribution and exposure rate from a 250-MBq 57Co flood source during quality control (QC) procedures as a function of source placement and measurement location. The optimum placement of the source to reduce the radiation dose to the nuclear medicine technologist during QC checks was determined. METHODS: Measurements of exposure rate were made inside and outside a camera room with the source positioned either above or below the camera head. The energy distribution of the photon field was examined at the same locations using a high-resolution gamma-ray spectrometer. Additional measures of exposure rate were made with the source at various distances from the camera face. RESULTS: The lowest exposure rates occurred when the source was lying directly on the face of the camera head. The exposure rates at locations inside the camera room increased by a factor of 4.3 +/- 3.0 when the source was placed on an imaging table below the camera head. This increase can be attributed to decreased shielding provided by the camera head. CONCLUSION: A large portion of the radiation dose received by technologists during QC checks is due to scattered radiation and x-rays produced by gamma-ray interactions within the camera. This dose can be reduced significantly if QC checks are performed with the flood source lying directly on the inverted gamma camera head rather than placing the flood source on an imaging table under the gamma camera. PMID- 10824620 TI - Is your technetium generator eluate sterile? AB - OBJECTIVE: This study was performed to assess the sterility of multidose 99mTc generator eluate vials at the end of a working day. METHODS: Expired 99mTc generator eluate vials were collected over a period of 10 wk and stored until the activity reached background. Four batches of 10 vials each were selected randomly and sent to an independent microbiology laboratory for sterility testing. RESULTS: No eluate showed any microbial growth after 14 d incubation in growth media. CONCLUSION: Retrospective sterility testing of 99mTc generator eluate confirmed the validity of our departmental protocol for radiopharmaceutical preparation. Sterility testing has become part of our quality control program. PMID- 10824621 TI - Standardized annotation of nuclear medicine images. AB - There are no generally accepted standards for annotating nuclear medicine images. This is a potential problem whenever hardcopies from other centers are being evaluated, reinterpreted or compared to actual images of the same patient. Proposals for image annotation are elaborated to support image evaluation by a third party. In this paper, examples are given of lung scintigraphy, thyroid scintigraphy, bone scintigraphy both in planar and SPECT techniques, renal function scintigraphy, myocardial perfusion scintigraphy, and PET. They are presented to stimulate discussion in the nuclear medicine community. PMID- 10824622 TI - Clinical pharmacology in the Middle Ages: principles that presage the 21st century. PMID- 10824623 TI - St John's Wort: effect on CYP3A4 activity. AB - BACKGROUND: St John's Wort is a widely used herbal product. Information regarding its potential for drug interactions is required for responsible treatment of patients using St John's Wort. CYP3A4 is a metabolic enzyme implicated in most clinically significant drug-drug interactions. OBJECTIVE: To determine the in vivo effect of reagent-grade St John's Wort extract on CYP3A4 activity through evaluation of urinary 6-beta-hydroxycortisol/cortisol ratios. METHODS: Thirteen subjects ranging in age from 18 to 25 years participated in this unblinded, multiple-dose, single-treatment before-after trial conducted in a university based pharmacokinetics and biopharmaceutics laboratory. Each subject ingested a 300-mg tablet of reagent-grade St John's Wort extract standardized to 0.3% hypericin three times a day for 14 days. Baseline and posttreatment CYP3A4 activity was assessed with the urinary 6-beta-hydroxycortisol/cortisol ratio after a 24-hour urine collection. RESULTS: The mean +/- SD urinary 6-beta hydroxycortisol/cortisol ratio significantly increased (P = .003) from a baseline value of 7.1 +/- 4.5 to 13 +/- 4.9. The mean +/- SD percentage increase was 114% +/- 95%, with a range from -25% to 259%. All but one subject had an increase in the ratio. CONCLUSIONS: Treatment with St John's Wort for 14 days resulted in significant increases in the urinary 6-beta-hydroxycortisol/cortisol ratio. This finding suggests that St John's Wort is an inducer of CYP3A4. PMID- 10824624 TI - Nicotine metabolism and elimination kinetics in newborns. AB - OBJECTIVE: To characterize the presence of and elimination kinetics of nicotine and its metabolites in newborns. METHODS: Blood samples from 13 newborns were collected during the first day of life and analyzed for nicotine and cotinine. Single daily urine samples were collected from nine newborns for up to 7 days and analyzed by gas chromatography-mass spectrometry for nicotine, cotinine, 3' hydroxycotinine, and their conjugates. NONMEM was used to determine population half-life values. RESULTS: Blood and urine data gave similar results for nicotine and cotinine elimination kinetics. The elimination half-life for nicotine was 11.2 hours (95% confidence interval [CI], 8.0 to 18.9) based on blood data and 9.0 hours (95% CI, 7.0 to 12.4) based on urine data. The elimination half-life for cotinine was 16.3 hours (95% CI, 12.4 to 23.9) based on blood data and was 22.8 hours (95% CI, 19.5 to 25.8) based on urine data. The elimination half-lives for the other metabolites were 13 hours for conjugated nicotine; 19.8 hours for conjugated cotinine; 18.8 hours for 3'-hydroxycotinine; and 19.4 hours for conjugated 3'-hydroxycotinine. The half-life of nicotine is three to four times longer in newborns than in adults, whereas the half-life of cotinine is similar in newborns and adults. CONCLUSIONS: In adults, CYP2A6 is the predominant enzyme responsible for the metabolism of both nicotine and cotinine. The prolonged elimination of nicotine but not of cotinine in the newborn compared with that in the adult may be a result of different newborn CYP2A6 enzymatic substrate specificity, low CYP2A6 activity with another enzyme that is primarily responsible for cotinine metabolism, or differences in tissue distribution. PMID- 10824625 TI - Significant interaction between the nonprescription antihistamine diphenhydramine and the CYP2D6 substrate metoprolol in healthy men with high or low CYP2D6 activity. AB - The prototype "classic" over-the-counter antihistamine diphenhydramine was shown to interact with the polymorphic P450 enzyme CYP2D6. This project was undertaken to investigate (1) whether diphenhydramine inhibits the biotransformation of the clinically relevant CYP2D6 substrate metoprolol in vitro and (2) whether this in vitro interaction results in a clinically significant pharmacokinetic and pharmacodynamic drug interaction in vivo. In vitro incubations were carried out with microsomes obtained from lymphoblastic cells transfected with CYP2D6 complementary deoxyribonucleic acid to determine the type and extent of inhibition. We then randomized 16 subjects with genetically determined high (extensive metabolizers) or low (poor metabolizers) CYP2D6 activity to receive metoprolol (100 mg) in the presence of steady-state concentrations of diphenhydramine or placebo. In vitro, diphenhydramine was a potent competitive inhibitor of metoprolol alpha-hydroxylation, exhibiting an inhibitory constant of 2 micromol/L and increasing the Michaelis-Menten constant of metoprolol sixfold. In vivo, diphenhydramine decreased metoprolol oral and nonrenal clearances twofold and metoprolol-->alpha-hydroxymetoprolol partial metabolic clearance 2.5 fold in extensive metabolizers (all P < .05) but not in poor metabolizers (P > .2). Although the hemodynamic response to metoprolol was unaltered by diphenhydramine in poor metabolizers (P > .05), metoprolol-related effects on heart rate, systolic blood pressure, and Doppler-derived aortic blood flow peak velocity were more pronounced and lasted significantly longer in extensive metabolizers receiving diphenhydramine compared with poor metabolizers and extensive metabolizers receiving placebo. We conclude that diphenhydramine inhibits the metabolism of metoprolol in extensive metabolizers, thereby prolonging the negative chronotropic and inotropic effects of the drug. Clinically relevant drug interactions may occur between diphenhydramine and many CYP2D6 substrates, particularly those with a narrow therapeutic index. PMID- 10824626 TI - Investigations of the metabolites of the trypanocidal drug melarsoprol. AB - BACKGROUND: Melarsoprol remains the first-choice drug for trypanosomiasis (human African sleeping sickness). To contribute to the sparse pharmacologic data and to better understand the cause of the frequent serious adverse reactions, we investigated the metabolism of this 50-year-old organoarsenic compound. RESULTS: The half-life of melarsoprol determined by HPLC was <1 hour compared with 35 hours determined by bioassay and atomic absorption spectroscopy, indicating the existence of active metabolites. One metabolite, melarsen oxide, was identified by ultraviolet HPLC after incubation of melarsoprol with microsomes. The maximum plasma concentration of melarsenoxide was reached 15 minutes after administration; the clearance was 21.5 mL/min/kg and the half-life of free melarsen oxide was 3.9 hours. Either melarsen oxide or a yet-undiscovered active metabolite is irreversibly bound to proteins, as shown by ultrafiltration, precipitation experiments, and atomic absorption spectroscopy. Because of the poor pharmaceutical properties of melarsoprol, the therapeutic potential of melarsen oxide was investigated. In a rodent model of acute infection, 20 of 20 mice were cured (0.1 to 1 mg/kg intravenously or 2.2 mg/kg intraperitoneally). In a rodent model of central nervous system infection, five of six mice survived for more than 180 days (5 mg/kg intravenously), indicating a sufficient melarsen oxide penetration across the blood-brain barrier. CONCLUSION: The prospects for the future of trypanosomiasis treatment are deplorable. Investigations on the improvement of the use of the old drugs are therefore required. The results of this study may build a basis for further research on the cause of severe adverse reactions. PMID- 10824627 TI - Population pharmacokinetics and pharmacodynamics of TS-943 for selective nonpeptide platelet glycoprotein IIb/IIIa receptor antagonist in normal healthy subjects. AB - The pharmacokinetics and pharmacodynamics of TS-943 were evaluated with use of NONMEM in 36 healthy male subjects after constant infusion of five different single-dose regimens. Population analysis showed the plasma concentration-time profiles of TS-943 to be best-fit characterized by a two-compartment open model with constant infusion and first-order elimination. The pharmacodynamic model that best fitted the platelet aggregation was a sigmoid Emax model. The final estimates for baseline effect, 50% inhibitory concentration (IC50), and the Hill coefficient were 79.4%, 23.4 ng/mL and 1.63, respectively. The maximum effect (Emax) was fixed at 80% (submaximal aggregation response). In addition, correlations between TS-943 plasma concentration and extension of template bleeding time were examined by fitting with an exponential model. The model estimates that the TS-943 plasma concentration necessary to double template bleeding time is approximately 63 ng/mL (ie, 2.7-fold greater than the IC50). The population approaches for pharmacokinetic-pharmacodynamic investigation can be useful for the analysis of concentration-effect relationships and concentration adverse event relationships for a platelet glycoprotein IIb/IIIa receptor antagonist. PMID- 10824628 TI - Pharmacokinetic interaction between verapamil and almotriptan in healthy volunteers. AB - OBJECTIVE: To assess the interaction between almotriptan, a 5-HT1B/1D-receptor agonist used to treat migraine, and verapamil, an agent for migraine prophylaxis. METHODS: Twelve healthy volunteers received the following treatments in a crossover design: (1) 120-mg sustained-release verapamil tablet twice daily for 7 days and one 12.5-mg almotriptan tablet on day 7 and (2) one 12.5-mg almotriptan tablet alone on day 7. Serial plasma and urine samples were obtained on day 7. Almotriptan plasma concentrations were determined by liquid chromatography-tandem mass spectrometry; urine samples were analyzed by ultraviolet HPLC. Safety measures included blood pressure and pulse measurements, electrocardiography, and adverse event monitoring. Statistical comparisons of pharmacokinetic parameters and vital sign data were made by ANOVA. RESULTS: Mean almotriptan peak concentration and area under the plasma concentration-time curve were significantly higher and volume of distribution and oral clearance were significantly lower after coadministration of almotriptan and verapamil compared with administration of almotriptan alone. The magnitudes of these differences were approximately 20%. Renal clearance was unaffected by verapamil coadministration. No significant effects of treatment on blood pressure or pulse were detected, with the exception of sitting systolic blood pressure at 2 hours after administration. However, the difference in mean change from baseline at this time point was only 8 mm Hg. CONCLUSIONS: Verapamil modestly inhibited almotriptan clearance to a degree consistent with the modest contribution of CYP3A4 to almotriptan metabolism. This observation and the lack of effect of verapamil on the tolerability to almotriptan administration suggest that no reduction of the almotriptan dose is warranted. PMID- 10824629 TI - The influence of reduced liver blood flow on the pharmacokinetics and pharmacodynamics of recombinant tissue factor pathway inhibitor. AB - OBJECTIVES: Recombinant tissue factor pathway inhibitor (rTFPI) has been shown to be an effective treatment in animal models of sepsis and is under investigation for human use. Reduced liver blood flow during septic shock may substantially alter the pharmacokinetics of rTFPI because clearance of rTFPI approaches liver blood flow. The aim of this study was to examine the effect of exercise-induced reduction in liver blood flow on the pharmacokinetics and pharmacodynamics of rTFPI. METHODS: This was a two-way, open-label, randomized crossover study in eight healthy male volunteers. The subjects in both treatment groups received a continuous intravenous infusion of rTFPI (0.2 mg/kg/h) concurrently with intravenous sorbitol (50 mg/min) for 4 hours. Sorbitol was used as a biomarker for liver blood flow. The subjects were randomized to remain supine or to exercise on a bicycle ergometer for 30 minutes starting at the beginning of the third hour of the infusion. RESULTS: Exercise reduced liver blood flow (mean +/- SEM) from 1.44 +/- 0.06 L/min to 0.40 +/- 0.03 L/min. The average clearance of rTFPI decreased from 0.73 +/- 0.04 L/min in the supine position to 0.25 +/- 0.02 L/min during exercise. This decrease in rTFPI clearance resulted in an 80% (95% confidence interval [CI], 60% to 102%) increase in plasma rTFPI levels during exercise. The average maximal prothrombin time and activated partial thromboplastin time values during exercise were 1.4 (95% CI, 0.4 to 2.5) and 4.4 (95% CI, 2.7 to 6.1) seconds higher compared with the supine steady-state level. CONCLUSIONS: Reduction in liver blood flow by exercise markedly increased rTFPI concentrations and induced a slight but variable prothrombin time and activated partial thromboplastin time increase at the rTFPI dose studied. PMID- 10824630 TI - Enzyme induction in the elderly: effect of rifampin on the pharmacokinetics and pharmacodynamics of propafenone. AB - OBJECTIVE: A clinical study on enzyme induction in elderly subjects was performed by investigation of the effect of rifampin (INN, rifampicin) on propafenone disposition. Propafenone was chosen as a model drug because of its complex metabolism that permits the simultaneous in vivo assessment of induction of phase 1 and phase 2 pathways. METHODS: Six extensive metabolizers of CYP2D6 (age, 70.5 +/- 3.5 years) ingested 600 mg rifampin once daily for 9 consecutive days. One day before the first rifampin dose and on the day of the last rifampin dose, each elderly individual received a single intravenous infusion of 70 mg unlabeled propafenone and received a single oral dose of 300 mg deuterated propafenone 2 hours later. Pharmacokinetics and pharmacodynamics of propafenone were compared before and during induction. RESULTS: Maximum QRS prolongation after oral propafenone was decreased significantly by rifampin (18% +/- 5% versus 6% +/- 3%; P < .01). There were no substantial differences in pharmacokinetics and pharmacodynamics of intravenous propafenone during induction. However, bioavailability of propafenone dropped from 30% +/- 24% to 4% +/- 3% (P < .05). After oral propafenone was administered, clearances through N-dealkylation (6 +/- 3 mL/min versus 26 +/- 16 mL/min; P < .05) and glucuronidation (178 +/- 75 mL/min versus 739 +/- 533 mL/min; P < .05), but not 5-hydroxylation, were increased by rifampin, indicating substantial enzyme induction. CONCLUSIONS: Both phase 1 and phase 2 pathways of propafenone metabolism were induced by rifampin in elderly subjects, resulting in a clinically relevant drug interaction. PMID- 10824631 TI - Mechanism of cardiotoxicity of halofantrine. AB - OBJECTIVES AND METHODS: To further evaluate the scope and mechanism of potential cardiotoxicity associated with the antimalarial drug halofantrine, case reports submitted to the US Food and Drug Administration Spontaneous Reporting System were examined. Because halofantrine was associated with electrocardiographic prolongation of the QT interval and ventricular arrhythmias, in vitro cardiac electrophysiologic studies (isolated perfused cardiac model and isolated ventricular myocytes) were conducted to test the hypothesis that halofantrine or its metabolite is responsible for cardiotoxicity. RESULTS: Although it is difficult to ascertain causality and to estimate overall incidence, a significant number of adverse events related to the cardiovascular system were reported, including QT interval prolongation, life-threatening arrhythmias, and sudden death. The effect of halofantrine and its active metabolite (N desbutylhalofantrine) on repolarization were examined in an isolated perfused heart model. Results indicate that halofantrine was able to prolong the QT interval, whereas N-desbutylhalofantrine had minimal effect on the QT interval relative to baseline. In an attempt to further elucidate the mechanism of QT interval prolongation, the effects of racemic halofantrine, its stereoisomers, and N-desbutylhalofantrine on repolarizing currents in isolated ventricular myocytes were studied with use of patch-clamp techniques. Halofantrine produced a stereoselective block of the delayed rectifier potassium channel in isolated feline myocytes. CONCLUSIONS: These results indicate that halofantrine is similar to quinidine and class III antiarrhythmics in its ability to prolong repolarization. We conclude that high plasma concentrations of halofantrine should be avoided, especially in women, and that N-desbutylhalofantrine may have potential as a safer antimalarial drug. PMID- 10824632 TI - Even low-dose aspirin inhibits arachidonic acid-induced vasodilation in heart failure. AB - BACKGROUND: There is some evidence that aspirin may be harmful to patients with congestive heart failure treated with angiotensin-converting enzyme (ACE) inhibitors, but there has never been any direct examination of the vascular effects of aspirin in these patients. We sought to determine whether there is an arachidonic acid-dependent vasodilator pathway in resistance arteries in humans, whether it is affected by congestive heart failure, and whether it is inhibited by low-dose aspirin. METHODS: A locally active dose of arachidonic acid was infused into the nondominant brachial artery while forearm blood flow was measured by venous occlusion plethysmography in 10 healthy subjects in a control group and 15 patients with congestive heart failure treated with ACE inhibitor. Patients with congestive heart failure were studied after administration of 0 mg, 75 mg, and 300 mg aspirin for 14 days. RESULTS: Arachidonic acid produced progressive and incremental vasodilation (up to 64%). There was no significant difference between patients and healthy control subjects studied after administration of 0 mg aspirin. In patients, however, administration of 75 mg and 300 mg aspirin inhibited mean vasodilation by 55% and 59%, respectively. CONCLUSIONS: There is an arachidonic acid-dependent vasodilator pathway in humans. This pathway is not significantly affected by congestive heart failure. It is significantly inhibited by even low-dose aspirin therapy. These results imply that even the very lowest dose of aspirin in common use for cardioprotection has potentially detrimental vasoconstrictor effects. PMID- 10824633 TI - Physostigmine reversal of midazolam-induced electroencephalographic changes in healthy subjects. AB - OBJECTIVE: Midazolam is a water-soluble benzodiazepine. Flumazenil is a potent antagonist of midazolam-induced sedation. Physostigmine has also been shown to reverse benzodiazepine sedation in anecdotal reports. The aim of this study was to quantitatively characterize the reversal of midazolam-induced changes in electroencephalogram (EEG) by physostigmine compared to flumazenil and placebo. METHODS: Twelve healthy male subjects received 5 mg midazolam as an intravenous infusion over 15 minutes. Fifteen minutes after the end of infusion, single doses of either 0.4 mg flumazenil, 0.5 mg physostigmine, or placebo (physiologic saline solution) were administered as intravenous injections in a randomized crossover fashion. Midazolam serum concentrations were measured using liquid chromatography tandem mass spectrometry. The time from the start of injection until awakening was noted and the EEG was measured. RESULTS: Four subjects were excluded from further pharmacokinetic and pharmacodynamic analysis because no midazolam-induced changes on EEG alpha power could be observed in each of the three study periods. The pharmacokinetics of midazolam were not influenced by flumazenil or physostigmine. Midazolam induced a decrease in EEG alpha power (7.50 to 11.25 Hz) compared with baseline (P < .05). After injection of flumazenil and physostigmine, an increase in EEG alpha power was observed, whereas placebo did not affect alpha power. Subjects opened their eyes 25.2 +/- 1.1 minutes after the placebo injection was begun, whereas subjects awoke after 6.2 +/- 2.7 minutes and 15.4 +/- 3.4 minutes after they received flumazenil and physostigmine, respectively (mean +/- SEM; P < .001). CONCLUSION: Physostigmine and flumazenil antagonized midazolam-induced sedation. This suggests that a reversible central anticholinergic mechanism may be involved in the sedative action of midazolam. PMID- 10824634 TI - Human cardiac beta1- or beta2-adrenergic receptor stimulation and the negative chronotropic effect of low-dose pirenzepine. AB - OBJECTIVES: The M1-muscarinic receptor antagonist pirenzepine in low doses (<1 mg intravenously) decreases heart rate. We investigated whether these effects of pirenzepine differ in volunteers with activated cardiac beta1-adrenergic receptors versus activated cardiac beta2-adrenergic receptors. METHODS: In 17 male volunteers (25 +/- 1 years) we studied effects of pirenzepine infusion (0.5 mg intravenous bolus followed by continuous infusion of 0.15 microg/kg/min) on heart rate and heart rate-corrected duration of electromechanical systole (QS2c, as a measure of inotropism) that had been stimulated by activation of cardiac beta1-adrenergic receptors (bicycle exercise in the supine position for 60 minutes at 25 W) or cardiac beta2-adrenergic receptors (continuous intravenous infusion of 100 ng/kg/min terbutaline). RESULTS: Bicycle exercise and terbutaline infusion significantly increased heart rate and shortened QS2c. When pirenzepine was infused 20 minutes after the beginning of the exercise or terbutaline infusion, heart rate decreased in both settings by approximately the same extent (approximately -10 to -14 beats/min), although exercise and terbutaline infusion continued; however, QS2c was not affected. Pirenzepine (0.05 to 1 mg intravenous bolus)-induced decrease in heart rate was abolished after 6 days of transdermal scopolamine treatment of volunteers. CONCLUSIONS: Low-dose pirenzepine decreased heart rate by muscarinic receptor stimulation, because this was blocked by scopolamine. Moreover, low-dose pirenzepine did not differentiate between cardiac beta1- or beta2-adrenergic receptor stimulation; however, low-dose pirenzepine did not affect cardiac contractility as measured by QS2c. Low-dose pirenzepine therefore exerted a unique pattern of action in the human heart: it decreased heart rate (basal and beta1- and/or beta2-adrenergic receptor-stimulated) without affecting contractility. PMID- 10824635 TI - In vivo beta3-adrenergic stimulation of human thermogenesis and lipid use. AB - OBJECTIVE: To investigate the role of the human beta3-adrenergic receptor in in vivo isoproterenol (INN, isoprenaline)-induced thermogenesis and lipid use. METHODS: Eight male volunteers participated in two studies. In the first study subjects received oral dosages of 2.5, 7.5, 15, and 40 mg nadolol or propranolol (both beta1- and beta2-adrenergic receptor antagonists) at random, after which isoproterenol (beta1-, beta2-, and beta3-adrenergic receptor agonist) was infused in an individually determined dosage (range, 19 to 35 ng/kg x min) that increased energy expenditure by 25% without pretreatment. In the second study, 50, 100, and 200 ng/kg x min isoproterenol or saline solution were infused after pretreatment with 80 mg nadolol. In both studies energy expenditure and respiratory exchange ratio were measured by indirect calorimetry and, at the end of each infusion period, blood samples were taken and tremor score (only first study), heart rate, and blood pressure were measured. RESULTS: In the first study, nadolol or propranolol in doses < or =40 mg could not fully block beta1-adrenergic receptor mediated increases in heart rate and systolic blood pressure. Propranolol in doses < or =7.5 mg could not fully block the beta2-adrenergic receptor-mediated increase in tremor score during isoproterenol infusion. The increases found in thermogenesis and lipid use could therefore be explained by concomitant beta1- and beta2-adrenergic stimulation. In the second study, isoproterenol infusion induced a significant increase in heart rate, but no increases in thermogenesis and lipid use were found compared with infusion of saline solution. CONCLUSION: No evidence could be found for a beta3-adrenergic receptor-mediated increase in human thermogenesis and lipid use during isoproterenol infusion after pretreatment with nadolol or propranolol. PMID- 10824636 TI - Relationship of paroxetine disposition to metoprolol metabolic ratio and CYP2D6*10 genotype of Korean subjects. AB - OBJECTIVE: To evaluate the relationship between the metabolic ratio (MR) of metoprolol, CYP2D6*10B genotype, and the disposition of paroxetine in Korean subjects. METHODS: A single 40-mg dose of paroxetine was administered orally to one poor metabolizer and 15 healthy subjects recruited from 223 Korean extensive metabolizers whose phenotypes were predetermined by use of the metoprolol MR. Genotypes were determined by allele-specific polymerase chain reaction and the GeneChip microarray technique. Pharmacokinetic parameters were estimated from plasma concentrations of paroxetine for more than 240 hours after the oral dose. RESULTS: The oral clearance and area under the plasma concentration versus time curve (AUC) of paroxetine were best described by a nonlinear relationship with metoprolol MR at correlation coefficients of 0.82 and 0.91, respectively (P < .05). Nine extensive metabolizer who were either homozygous or heterozygous for CYP2D6*10B had significantly lower oral clearance values of paroxetine than six extensive metabolizers with CYP2D6*1/*1. The AUC of paroxetine in subjects who were homozygous for CYP2D6*10B (666.4 +/- 169.4 ng/mL x h) was significantly greater than that of subjects who were homozygous for the wild type (194.5 +/- 55.9 ng/mL x h). Unexpectedly, the average AUC of subjects who were heterozygous for CYP2D6*10B was greater with wide variation (789.8 +/- 816.9 ng/mL x h) than that of subjects who were homozygous CYP2D6*10B/*10B mainly because of two atypical subjects whose metoprolol MR was not associated with the CYP2D6*10B genotype and who showed greater AUC and lower oral clearance than subjects with homozygous CYP2D6*10B. CONCLUSIONS: The CYP2D6 activity measured by metoprolol MR was a strong predictor of paroxetine disposition in Korean extensive metabolizers. In general, the extensive metabolizers with the CYP2D6*10B allele seemed to have higher plasma concentrations of paroxetine than extensive metabolizers with the wild-type CYP2D6 genotype. However, quantitative prediction of paroxetine disposition from the CYP2D6*10B genotype alone was not perfect because several Korean extensive metabolizers had metoprolol MRs that were not associated with the genotype. PMID- 10824637 TI - Erythromycin breath test. PMID- 10824638 TI - Duplicate publishing and the least publishable unit. PMID- 10824639 TI - Response of males and females to high-force eccentric exercise. AB - It has previously been shown that females incur less muscle damage than males after strenuous exercise, but limited data are available for humans. To determine possible differences between the sexes in humans, the response to high-force eccentric exercise was examined in a large sample of women (n = 83) and men (n = 82). The participants performed a bout of eccentric exercise of the elbow flexors consisting of 70 maximal repetitions. Isometric strength, resting elbow angle and muscle soreness were measured before, immediately after (except soreness) and then daily for 7 days after exercise. There was a significant loss in strength among both groups (69% for women and 63% for men) (P < 0.01) immediately after exercise; at 168 h post-exercise, women still had a 27% strength loss and men had a 24% strength loss. No significant difference in strength loss or recovery rate was found between men and women. Soreness reached peak values 32-48 h post exercise (P < 0.01), with no significant difference between men and women. Range of motion decreased significantly until 3 days after exercise (14.6 degrees or 0.255 rad loss for women; 12.2 degrees or 0.213 rad loss for men) (P < 0.01); at 168 h post-exercise, the women and men still showed a loss of 4.8 degrees (0.084 rad) and 4.0 degrees (0.07 rad), respectively. There was a significant interaction of sex x time (P < 0.01); a post-hoc test indicated that the women experienced a greater loss in range of motion at 72 h than men and this difference was maintained to 168 h post-exercise (P < 0.01). Thus, our results do not support the contention that women have a lower response to eccentric exercise than men. PMID- 10824640 TI - Force production in the rugby union scrum. AB - In this study, we examined the relationship between anthropometric, strength and power characteristics of rugby forwards, their body position when scrummaging, and their ability to apply force when scrummaging. Force applied to an instrumented scrum machine was measured for 56 players, both individually and as scrum packs. Measurements of body position for individuals were made by digitizing videotape records of the trials. Forty players subsequently had their anthropometry assessed and completed several strength and power tests. Body mass, each component of somatotype, maximal anaerobic power developed on a cycle ergometer, and isokinetic knee extension strength correlated significantly with individual scrummaging force. A regression model (P < 0.001) including body mass, mesomorphy, maximal anaerobic power and hip angle while in the scrummaging position accounted for 45% of the variance in individual scrummaging force. The packs that produced the largest scrummaging forces were, in general, characterized by a greater pack force to sum of individual force ratio than the packs producing lower forces. Our results emphasize the need for a scrum pack to develop technique and coordination as a unit to maximize scrummaging force. PMID- 10824641 TI - Long-axis rotation: the missing link in proximal-to-distal segmental sequencing. AB - Most assessments of segmental sequencing in throwing, striking or kicking have indicated a proximal-to-distal sequencing of end-point linear speeds, joint angular velocities, segment angular velocities and resultant joint moments. However, the role of long-axis rotations has not been adequately quantified and located in the proximal-to-distal sequence. The timing and importance of upper arm internal-external rotation and pronation-supination in the development of racquet head speed have been examined in the tennis serve and squash forehand drive and considered in relation to conventional concepts of proximal-to-distal sequencing. Both long-axis rotations reached their peak angular speeds late in both strokes, typically after shoulder flexion-extension, shoulder abduction adduction and elbow extension. These results clarify and confirm the importance of upper limb long-axis rotations in the production of racquet head speed. It appears that traditional proximal-to-distal sequencing concepts are inadequate to describe accurately the complexity of the tennis serve or squash forehand drive. It is essential to consider upper arm and forearm longitudinal axis rotations in explaining the mechanics of these movements and in developing coaching emphases, strength training schedules and injury prevention programmes. PMID- 10824642 TI - Effect of exercise and nasal splinting on static and dynamic measures of nasal airflow. AB - The main aim of this study was to assess the separate and combined effects of exercise and nasal splinting on static and dynamic measures of nasal airflow. In a randomized crossover design, 12 healthy participants (6 men, 6 women) performed static and dynamic spirometric nasal airflow assessment tests, with or without nasal splinting (Breathe-Right), before and after a maximal oxygen uptake (VO2max) treadmill test. At least 7 days later, the VO2max, and nasal airflow tests were repeated. The results showed that the measured variables were not significantly different with and without nasal splinting. We conclude that the absence of significantly enhanced nasal patency observed for nasal splinting and after exercise suggest that these factors have a minimal impact on nasal airflow volume and rate. PMID- 10824643 TI - Prediction of fat and fat-free mass in male athletes using dual X-ray absorptiometry as the reference method. AB - The ability of bioelectrical impedance analysis and anthropometry to predict fat mass and fat-free mass was compared in a sample of 82 male athletes from a wide variety of sports, using dual-energy X-ray absorptiometry (DXA) as the reference method. The percent fat measured by DXA was 10.9+/-4.9% (mean +/- s), and fat mass was predicted with a standard error of the estimate of 1.7 kg for skinfolds and 2.8 kg for bioelectrical impedance analysis (P < 0.001). Fat-free mass was predicted with a standard error of the estimate of 1.7 kg for anthropometry and 2.6 kg for bioelectrical impedance analysis (P < 0.001). Regression of various individual skinfolds and summed skinfolds, to examine the effect of skinfold selection combinations by stepwise regression, produced an optimal fat mass prediction using the thigh and abdominal skinfold sites, and an optimal fat-free mass prediction using the thigh, abdominal and supra-ilium sites. These results suggest that anthropometry offers a better way of assessing body composition in athletes than bioelectrical impedance analysis. Applying the derived equations to a separate sample of 24 athletes predicted fat and fat-free mass with a total error of 2.3 kg (2.9%) and 2.2 kg (2.7%), respectively. Combining the samples introduced more heterogeneity into the sample (n = 106), and the optimal prediction of fat mass used six skinfolds in producing a similar standard error of the estimate (1.7 kg), although this explained a further 4% of the variation in DXA-derived fat. Fat-free mass was predicted best from four skinfolds, although the standard error of the estimate and coefficient of determination were unchanged. PMID- 10824644 TI - Examination of the psychometric properties of the perceived motivational climate in sport questionnaire-2 in a sample of female athletes. AB - We undertook two studies to determine the validity and reliability of the revised Perceived Motivational Climate in Sport Questionnaire (PMCSQ-2). In Study 1, 201 female athletes (mean age 16.4 years) were administered the initial version of the PMCSQ-2 and a measure of reported tension and pressure experienced in sport. Exploratory principal component analysis suggested that the PMCSQ-2 contained two higher-order scales (Task-Involving and Ego-Involving climates), each with three subscales (Task: Cooperative Learning, Effort/ Improvement, Important Role; Ego: Intra-Team Member Rivalry, Unequal Recognition, Punishment for Mistakes). In Study 2, 385 female volleyball players (mean age 15.2 years) completed the PMCSQ 2, the Intrinsic Motivation Inventory and a measure of Team Satisfaction. Confirmatory factor analysis was applied to six competing models. The oblique six factor model and oblique hierarchical model provided comparable fit to the data. Acceptable fit was reached based on model respecification. Across Studies 1 and 2, internal consistency was found to be acceptable for the higher-order scales and subscales (with the exception of the Intra-Team Member Rivalry subscale). We found evidence for the concurrent validity of the instrument. PMID- 10824645 TI - Contributions of local and central sensations to the perception of exertion during cycling: effects of work rate and cadence. AB - There is evidence to suggest that perception of exertion during exercise is based on both local and central sensations. The aim of the present experiment was to determine the relative contributions of different sensations to overall perceived exertion during cycling. Eighteen trained cyclists pedalled on a cycle ergometer for 4 min at each of three work rates (100, 150 and 200 W) and cadences (50, 70 and 90 rev x min(-1)). At the end of each bout, they used Borg's category-ratio (CR-10) scale to rate their overall perceived exertion, leg muscle pain, knee pain, breathlessness and heart beat intensity. The results indicated that cadence only influenced local sensations (muscle pain and knee pain), which were significantly higher at slower pedalling rates. Neither overall perceived exertion nor central sensations (breathlessness and heart beat intensity) were significantly affected by cadence. In contrast, increases in work rate were associated with higher ratings for all sensations. Further analyses revealed that variations in these overall ratings of perceived exertion as a function of work rate were accounted for by variations in ratings of muscle pain and breathlessness. The general implication is that perceived exertion during cycling derives from a combination of muscle and respiratory sensations. PMID- 10824646 TI - Publication bias and the integrity of psychiatry research. PMID- 10824647 TI - Interferons, serotonin and neurotoxicity. AB - BACKGROUND: Interferons are a class of cytokines profoundly affecting immune function. Several interferons are now synthesized and used clinically, notably for viral diseases and cancer. In addition to their desired immune effects, interferons cause a number of toxicities, including prominent effects on the nervous system. METHODS: This literature review focused on the incidence of depression associated with interferon treatment. Possible neurochemical mechanisms and remedial strategies were also considered. RESULTS: Interferon treatment, particularly with the alpha subtype, is unquestionably linked with depression, but the strength of association is uncertain because of erratic ascertainment and pretreatment co-morbidity. A likely pathogenic mechanism has been described, involving interferon suppression of serotonin synthesis. Controlled treatment trials of interferon-induced depression are not yet available. CONCLUSIONS: Neurotoxicity substantially limits the use of interferons. At least some of the risk of depression appears to derive from their anti-serotonergic effects, consistent with the large body of evidence pointing to a general link between serotonin and affective illness. Vigilant detection and aggressive treatment of depression is necessary to optimize interferon treatment of many patients. PMID- 10824648 TI - Urban-rural mental health differences in great Britain: findings from the national morbidity survey. AB - BACKGROUND: Studies of urban-rural differences in prevalence of non-psychotic mental disorder have not given consistent findings. Such differences have received relatively little study in Great Britain. METHODS: Data from 9777 subjects in the Household Survey of the National Morbidity Survey of Great Britain were analysed for differences between urban, semi-rural and rural areas. Psychiatric morbidity was assessed by scores on the Revised Clinical Interview Schedule (CIS-R), together with alcohol dependence, drug dependence, receipt of treatment from general practitioners. Associations with other characteristics were examined by logistic regression. RESULTS: Urban subjects had higher rates than rural of CIS-R morbidity, alcohol dependence and drug dependence, with semi rural subjects intermediate. Urban subjects also tended to be members of more deprived social groups, with more adverse living circumstances and greater life stress, factors themselves associated with disorder. Urban-rural differences in alcohol and drug dependence were no longer significant after adjustment for these factors by logistic regression, and differences on CIS-R morbidity were considerably reduced. There were no differences in treatment. CONCLUSIONS: There are considerable British urban rural differences in mental health, which may largely be attributable to more adverse urban social environments. PMID- 10824649 TI - Parenting and adult mood, anxiety and substance use disorders in female twins: an epidemiological, multi-informant, retrospective study. AB - BACKGROUND: Although parenting has long been considered an important risk factor for subsequent psychopathology, most investigations of this question have studied a single informant, clinical populations, one or a few disorders and did not consider relevant covariates. METHODS: Three dimensions of parenting (coldness, protectiveness and authoritarianism) were measured by combining the retrospective reports from adult female twins, their co-twins, and their mothers and fathers. We assessed by personal interview, lifetime history in the twins of eight common psychiatric and substance abuse disorders and a range of predictors of parenting. Analyses were performed using logistic regression. RESULTS: Examined individually, high levels of coldness and authoritarianism were modestly but significantly associated with increased risk for nearly all disorders, while the impact of protectiveness was more variable. These associations declined modestly when putative predictors of parenting were added as covariates. Maternal and paternal parenting were equally associated with outcomes in adult daughters. When coldness, protectiveness and authoritarianism were examined together, nearly all significant associations were seen solely with coldness. Few significant interactions were found between maternal and paternal parenting or between coldness, protectiveness and authoritarianism. The shared experience of these three dimensions of parenting predicts a quite small correlation in liability to these disorders in dizygotic twin pairs (e.g. r < 0.04). CONCLUSION: In women, parenting behaviour, especially levels of coldness, is probably causally related to risk for a broad range of adult psychiatric disorders. The impact of parenting on substance use disorders may be largely mediated through their co-morbidity with major depression, phobias and generalized anxiety disorder. In general population samples, the association of poor parenting with psychiatric illness is modest, largely non-specific and explains little of the observed aggregation of these disorders in families. PMID- 10824650 TI - The capacity of people with a 'mental disability' to make a health care decision. AB - BACKGROUND: Based on the developing clinical and legal literature, and using the framework adopted in draft legislation, capacity to make a valid decision about a clinically required blood test was investigated in three groups of people with a 'mental disability' (i.e. mental illness (chronic schizophrenia), 'learning disability' ('mental retardation', or intellectual or developmental disability), or, dementia) and a fourth, comparison group. METHODS: The three 'mental disability' groups (N = 20 in the 'learning disability' group, N = 21 in each of the other two groups) were recruited through the relevant local clinical services; and through a phlebotomy clinic for the 'general population' comparison group (N = 20). The decision-making task was progressively simplified by presenting the relevant information as separate elements and modifying the assessment of capacity so that responding became gradually less dependent on expressive verbal ability. RESULTS: Compared with the 'general population' group, capacity to make the particular decision was significantly more impaired in the 'learning disability' and 'dementia' groups. Importantly, however, it was not more impaired among the 'mental illness' group. All the groups benefited as the decision-making task was simplified, but at different stages. In each of the 'mental disability' groups, one participant benefited only when responding did not require any expensive verbal ability. CONCLUSIONS: Consistent with current views, capacity reflected an interaction between the decision-maker and the demands of the decision-making task. The findings have implications for the way in which decisions about health care interventions are sought from people with a 'mental disability'. The methodology may be extended to assess capacity to make other legally-significant decisions. PMID- 10824651 TI - Assessing the capacity of people with intellectual disabilities to be witnesses in court. AB - BACKGROUND: People with intellectual disabilities who have been victims or other witnesses of crime have had limited access to the criminal justice system, often on the basis of assumptions about their incapacity to be interviewed by the police and to give evidence in court. The aim of this study was to assess their capacity to be witnesses in court. METHODS: Forty-nine men and women with intellectual disabilities, all of whom were potential witnesses of ill-treatment, were assessed in order to provide advice, initially to the police, about their capacity to be interviewed for judicial purposes. The assessments included evaluations of each person's intellectual ability, memory, acquiescence, suggestibility, and their ability to explain concepts relating to the oath. RESULTS: Only 37 (76%) were able to complete the assessments. Most of those with a Full Scale IQ score of > or = 60 had a basic understanding of the oath, compared with only a third of those with IQ scores between 50 and 59, and none of those with IQ scores < 50. Nevertheless, some of the people who were unable to demonstrate an understanding of the oath did understand the words 'truth' and 'lie', especially when asked about these concepts in relation to concrete examples. CONCLUSIONS: While intellectual ability appears to be the best overall predictor of the capacity of people with intellectual disabilities to act as witnesses, confining witnesses to those who could explain the meaning of the oath would mean that a number of persons who might be interviewed by the police and subsequently appear in court could be excluded from the judicial process. PMID- 10824652 TI - Decreasing seasonal variation of births in schizophrenia. AB - BACKGROUND: Patients with schizophrenia have a winter-spring excess of births compared with the general population, the cause of which is unresolved. Fluctuations in the magnitude of the seasonal variation may provide clues to its aetiology. METHODS: All Finnish patients with schizophrenia born between 1950 and 1969 (N = 15892) were identified from two nationwide health-care registers. Their background demographic information was obtained from the Population Register Centre, which also provided monthly numbers of births in each municipality of Finland as multidimensional tables, with sex and year, month and place of birth as marginals. The incidence of schizophrenia was modelled using Poisson regression analysis, with sex, onset age, birth cohort, place of birth (urban/rural), trend and seasonal variation as explanatory variables. We also constructed a monthly time series and decomposed it into three components- seasonal, trend and remainder. RESULTS: Seasonal variation of births among patients born in the 1950s, especially between 1955 and 1959, was marked, but decreased among patients born in the 1960s. No interaction between place of birth or sex and seasonal variation was observed. The incidence was higher among the rural-born than the urban-born, but declined more slowly among the urban-born than the rural-born. CONCLUSIONS: The intensity of the factor causing the seasonal variation of births in schizophrenia may be decreasing. Urban birth may be emerging as a risk factor for schizophrenia in Finland, as elsewhere. PMID- 10824653 TI - A neuropsychological comparison of schizophrenia and schizophrenia-like psychosis of epilepsy. AB - BACKGROUND: The schizophrenia-like psychoses of epilepsy (SLPE) might represent a secondary form of schizophrenia in which the pathology is relatively confined to the temporal lobe. To test this possibility we have compared the neuropsychological profile of schizophrenia and SLPE. Our main hypothesis was that both psychotic groups would show deficits of temporal lobe function but that prefrontal impairment, as measured by tests of executive function, would be found only in the primary schi ophrenic group. METHODS: Four groups were studied: (1) patients with SLPE (N = 25); (2) patients with epilepsy but not psychiatric history (N = 24); (3) patients with schizophrenia (N = 22); and (4) healthy volunteers (N = 24). Neuropsychological testing comprised measures of pre-morbid IQ, current verbal and performance IQ, information processing, digit span, motor speed, verbal and visual learning and memory, verbal fluency, the Wisconsin Card Sorting Task, the Stroop test and the trail making task. RESULTS: Patients with schizophrenia and those with SLPE had almost identical neuropsychological profiles, with impairments of attention, episodic memory (verbal > visual) and executive function. The epileptic controls showed similar though less severe impairments of memory and of some tests of executive function. CONCLUSIONS: Our results do not support the hypothesis that the pathophysiology of SLPE and schizophrenia are distinct. While our findings suggest an important role for dominant temporal lobe abnormality in schizophrenia, both in its primary form and in that occurring in patients with epilepsy, they also implicate generalized cognitive impairment, manifest in particular as attentional deficits, in both forms of the disorder. PMID- 10824654 TI - The power and omnipotence of voices: subordination and entrapment by voices and significant others. AB - BACKGROUND: Cognitive therapy for psychotic symptoms often embraces self evaluative beliefs (e.g. self-worth) but whether and how such beliefs are related to delusions remains uncertain. In previous research we demonstrated that distress arising from voices was linked to beliefs about voices and not voice content alone. In this study we examine whether the relationship with the voice is a paradigm of social relationships in general, using a new framework of social cognition, 'ranking' theory. METHOD: In a sample of 59 voice hearers, measures of power and social rank difference between voice and voice hearer are taken in addition to parallel measures of power and rank in wider social relationships. RESULTS: As predicted, subordination to voices was closely linked to subordination and marginalization in other social relationships. This was not the result of a mood-linked appraisal. Distress arising from voices was linked not to voice characteristics but social and interpersonal cognition. CONCLUSION: This study suggests that the power imbalance between the individual and his persecutor(s) may have origins in an appraisal by the individual of his social rank and sense of group identification and belonging. The results also raise the possibility that the appraisal of voice frequency and volume are the result of the appraisal of voices' rank and power. Theoretical and novel treatment implications are discussed. PMID- 10824655 TI - Sensitivity to linguistic anomalies in spoken sentences: a case study approach to understanding thought disorder in schizophrenia. AB - BACKGROUND: As a group, positively thought-disordered (TD) schizophrenic patients are relatively impaired in their ability to use linguistic context to process sentences online (Kuperberg et al. 1998). This study investigates the heterogeneity in the use of linguistic context both between individual TD patients and within the individual patients as severity of thought disorder changes over time. METHODS: Seventeen TD schizophrenics performed an online word monitoring task on four separate occasions. In each patient, baseline reaction time (RTs) to target words in normal sentences were subtracted from RTs to target words in pragmatically-, semantically- and syntactically-violated sentences to obtain a measure of online sensitivity to each type of linguistic violation, and these were compared with normative data of a healthy volunteer and a non-TD schizophrenic control group. In addition, the co-variation of severity of thought disorder and sensitivity to linguistic context within all individual TD patients over the four testing sessions, was examined. RESULTS: There was marked heterogeneity between individual TD patients in their sensitivity to different types of linguistic violations: some were selectively insensitive to pragmatic violations, while others were insensitive to semantic and syntactic (subcategorization) violations. There was also an inverse relationship between severity of thought disorder and sensitivity to linguistic violations within individual patients over the four sessions. CONCLUSIONS: It is likely that a single cognitive deficit does not account for all types of schizophrenic thought disorder, but rather that there are multiple deficits affecting specific levels of linguistic processing. In these schizophrenic patients, impairment in the use of linguistic context was related to the state, rather than the trait, of thought disorder. PMID- 10824656 TI - Six-month and 12-month mental health outcome of medical and surgical patients admitted to general hospital. AB - BACKGROUND: We have recently reported a two-phase study on psychiatric morbidity in a sample of general hospital patients. This paper reports the results of the 6 month and 12-month follow-up of these patients. METHODS: The screening questionnaire was the GHQ-12. The main diagnostic instrument used in the second phase was the CIDI-PHC. All patients who had been interviewed with CIDI-PHC (N = 363) were followed-up and the baseline assessment was compared with the scoring on questionnaires administered in the 6-month postal enquiry and with the psychopathological status at 12-month, elicited with a telephone structured interview. RESULTS: Sixty-two and 87% of patients completed the 6- and 12-month follow-up assessment respectively. The first follow-up indicated no significant decrease in the level of symptoms. The 12-month follow-up interview showed that 23% of males and 40% of females had poor/mostly poor mental health. The logistic model showed that females with a definite ICD-10 diagnosis, admitted to a medical department, who had consumed psychotropic drugs in the previous year, had the most unfavourable outcome. The risk of a poor/mostly poor outcome steadily increased with the severity of the psychopathology during hospitalization. CONCLUSION: In medical and surgical general hospital patients the risk factors associated with a poor mental health outcome are similar to those found in primary care patients. Greater attention should be paid in assessing routinely mental health status of general hospital patients during hospitalization. PMID- 10824657 TI - Physical health and the onset and persistence of depression in older adults: an eight-wave prospective community-based study. AB - BACKGROUND: Poor physical health has long been recognized to be one of the most important risk factors for depression in older adults. Since many aspects of physical health can be targeted for improvement in primary care, it is important to know whether physical health problems predict the onset and/or the persistence of depression. METHODS: The study is based on a sample which at the outset consisted of 327 depressed and 325 non-depressed older adults (55-85) drawn from a larger random community-based sample in the Netherlands. Depression was measured using the Center for Epidemiologic Studies Depression scale (CES-D) at eight successive waves. RESULTS: From all incident episodes, the majority (57%) was short-lived. These short episodes could generally not be predicted by physical health problems. The remaining incident episodes (43%) were not short lived and could be predicted by poor physical health. Chronicity (34%) was also predicted by physical health problems. CONCLUSIONS: The study design with its frequent measurements recognized more incident cases than previous studies; these cases however did have a better prognosis than is often assumed. The prognosis of prevalent cases was rather poor. Physical health problems were demonstrated to be a predictor of both the onset and the persistence of depression. This may well have implications for prevention and intervention. PMID- 10824658 TI - Relationships between hostility and physiological coronary heart disease risk factors in young adults: the moderating influence of depressive tendencies. AB - BACKGROUND: We examined whether the relationships between hostility and physiological coronary heart disease (CHD) risk factors differ as a function of depressive tendencies (DT). METHODS: The participants were 672 randomly selected healthy young adults who self-reported their hostility (anger, cynicism, and paranoia) and DT. The physiological CHD risk factors studied were systolic blood pressure, diastolic blood pressure, body-mass index, serum high-density lipoprotein cholesterol, serum low-density lipoprotein cholesterol and serum triglycerides. RESULTS: We found that hostility was negatively associated with the physiological CHD risk factors among individuals exhibiting high DT while hostility was positively associated with, or unrelated to, the physiological risk factors among individuals showing low DT. The Hostility x DT interaction explained 2 to 5 % of the variance in the physiological parameters. CONCLUSION: The findings suggest that DT have a moderating influence on the relationships between hostility and CHD risk. Despite the established risk factor status of hostility, lack of anger and hostility, when combined with high DT, may represent the most severe exhaustion where the individual has given up. Disregard of this fact may explain some null findings in the research on hostility and CHD risk. PMID- 10824659 TI - Depression and social stress in Pakistan. AB - BACKGROUND: The high prevalence of depression in developing countries is not well understood. This study aimed to replicate the previous finding of a high prevalence of depression in Pakistan and assess in detail the associated social difficulties. METHOD: A two-phase survey of a general population sample in a Pakistani village was performed. The first-phase screen used the Personal Health Questionnaire (PHQ) and the self-rating questionnaire (SRQ). A one in two sample of high scorers and a one in three sample of the low scorers were interviewed using the Psychiatric Assessment Schedule (PAS) and Life Events and Difficulties Schedule (LEDS). RESULTS: A total of 259 people were screened (96% response rate). The second stage yielded 55 cases, of whom 54 had depressive disorder, and 48 non-cases. The adjusted prevalence of depressive disorders was 44-4% (95% CI 35.3 to 53.6): 25.5% in males and 57.5% in females. Nearly all cases had lasted longer than 1 year. Comparison of the cases and non-cases indicated that cases were less well educated, had more children and experienced more marked, independent chronic difficulties. Multivariate analysis indicated that severe financial and housing difficulties, large number of children and low educational level were particularly closely associated with depression. CONCLUSION: This study confirms the high prevalence of depressive disorders in Pakistan and suggests that this may be higher than other developing countries because of the high proportion of the population who experience social adversity. PMID- 10824660 TI - Diagnostic depressive symptoms of the mixed bipolar episode. AB - BACKGROUND: There is not yet consensus on the best diagnostic definition of mixed bipolar episodes. Many have suggested the DSM-III-R/-IV definition is too rigid. We propose alternative criteria using data from a large patient cohort. METHODS: We evaluated 237 manic in-patients using DSM-III-R criteria and the Scale for Manic States (SMS). A bimodally distributed factor of dysphoric mood has been reported from the SMS data. We used both the factor and the DSM-III-R classifications to identify candidate depressive symptoms and then developed three candidate depressive symptom sets. Using ROC analysis we determined the optimal threshold number of symptoms in each set and compared the three ROC solutions. The optimal solution was tested against the DSM-III-R classification for crossvalidation. RESULTS: The optimal ROC solution was a set, derived from both the DSM-III-R and the SMS, and the optimal threshold for diagnosis was two or more symptoms. Applying this set iteratively to the DSM-III-R classification produced the identical ROC solution. The prevalence of mixed episodes in the cohort was 13.9% by DSM-III-R, 20.2% by the dysphoria factor and 27.4% by the new ROC solution. CONCLUSIONS: A diagnostic set of six dysphoric symptoms (depressed mood, anhedonia, guilt, suicide, fatigue and anxiety), with a threshold of two symptoms, is proposed for a mixed episode. This new definition has a foundation in clinical data, in the proved diagnostic performance of the qualifying symptoms, and in ROC validation against two previous definitions that each have face validity. PMID- 10824661 TI - Do general practitioners' attitudes towards depression predict their clinical behaviour? AB - BACKGROUND: GPs' attitudes towards depression vary, as do their abilities to detect and manage it effectively. Associations between attitudes and clinical behaviour have not yet been demonstrated directly. We tested two hypotheses: (1) that questionnaire measures of GPs' confidence in identifying depression predict their ability to identify depression in their patients; and (2) that GPs who prefer antidepressants prescribe more than those who prefer psychotherapy. METHODS: Forty GPs in Liverpool and Manchester completed the Depression Attitude Questionnaire (DAQ) and were asked for prescribing (PACT) information. Attender surveys using the General Health Questionnaire (GHQ-12), in combination with GP ratings of patients' psychological status, generated indices for GPs' case identification, bias and accuracy. We tested associations between these indices and the four DAQ components, in particular GPs' confidence in diagnosis, across a total of 1436 patients. We also compared the DAQ component on attitudes to treatment with relevant PACT data. RESULTS: The DAQ assessment of GPs' ability to identify cases of depression bore no relationship to their observed ability, as measured by accuracy, bias, or identification indices. However, there were significant associations between observed diagnostic ability and: preference for psychotherapy; ease in managing depression; and, belief in successful treatment. PACT data were available for 26 (65%) GPs. There was an association between preference for antidepressants and prescription of SSRIs (rs 0.3981, P < 0.044), but not for overall antidepressant or tricyclic prescribing, or for dose of dothiepin. CONCLUSIONS: The DAQ measure of ease of identification is not valid when compared to actual clinical practice. The ability of GPs to identify depression may not be an independent variable, but may rather reflect other beliefs, attitudes and skills. This has considerable implications for educational interventions in primary care. PMID- 10824662 TI - Non-linear relationships in associations of depression and anxiety with alcohol use. AB - BACKGROUND: Many studies have demonstrated co-morbidity of alcohol abuse/dependence with mood and anxiety disorders but relatively little is known about anxiety and depression across the full continua of alcohol consumption and problems associated with drinking. METHODS: Participants from a general population sample (N = 2725) aged 18-80 years completed the Alcohol Use Disorders Identification Test (AUDIT) and four measures of negative affect (two depression and two anxiety symptom scales) included in a self-completion questionnaire. RESULTS: High consumption, AUDIT total score, and AUDIT problems score were associated with high negative affect scores in participants under 60 years old (ORs in the range 1.80-2.83). Graphical and statistical analyses using continuous measures of alcohol use/problems and negative affect identified non-linear relationships where abstainers and occasional drinkers, as well as heavy and problem drinkers, were at risk of high anxiety and depression levels. This pattern, however, was not found in those aged > or = 60 years. The U-shaped relationship was not an artefact of abstainers being typical of the general population in their distribution of negative affect. CONCLUSIONS: Studies of co morbidity should acknowledge the possibility of non-linear associations and employ both continuous and discrete measures. Abstainers, as well as heavy drinkers, are at increased risk of symptoms of depression and anxiety disorders. Psychosocial factors may play a role in the U-shaped relationship between alcohol consumption and mortality. PMID- 10824663 TI - The difference in patterns of motor and cognitive function in chronic fatigue syndrome and severe depressive illness. AB - BACKGROUND: Chronic fatigue syndrome (CFS) and major depressive disorder (MDD) share many symptoms and aetiological factors but may have different neurobiological underpinnings. We wished to determine the profile of the biological variables disturbed in CFS and MDD, and identify any critical factors that differentiate the disorders. METHODS: Thirty patients with CFS, 20 with MDD and 15 healthy controls matched group-wise for age and sex were recruited. Subjects were given a detailed battery of motor and cognitive tests, including measures of psychomotor speed, memory and maximal voluntary muscle contraction in both the morning and evening that were balanced to avoid order effects. RESULTS: CFS patients generally performed worse on cognitive tests than healthy controls, but better than patients with MDD. Both patient groups had markedly impaired motor function compared with healthy controls. MDD subjects showed a significantly greater diurnal improvement in maximal voluntary contraction than healthy controls. CONCLUSIONS: Patients with CFS and MDD show similarly substantial motor impairment, but cognitive deficits are generally more marked in MDD. Diurnal changes in some functions in MDD may differentiate the disorder from CFS. PMID- 10824664 TI - Monoamine oxidase: associations with alcohol dependence, smoking and other measures of psychopathology. AB - BACKGROUND: Many reports have appeared on associations between platelet monoamine oxidase (MAO) activity and susceptibility to psychiatric conditions; principally alcohol dependence but also conduct disorder, other drug use and depression. Recently, it has become apparent that MAO activity is inhibited by some component of cigarette smoke, and smokers have low platelet MAO activity. Since the prevalence of smoking is higher in many of the conditions in which MAO has been implicated, the MAO susceptibility associations may be partly, or entirely, false. METHODS: We have measured platelet MAO in 1551 subjects, recruited from the Australian NHMRC Twin Registry, who have provided information on alcohol use and dependence, smoking, conduct disorder, depression, attempted suicide, panic disorder and social phobia. RESULTS: Current smoking reduced platelet MAO activity in a significant and dose-related manner, with no evidence of lower MAO in ex-smokers or in non-smoking subjects with co-twins who smoked. Alcohol use and lifetime DSM-III-R alcohol dependence history were not associated with MAO activity when smoking was taken into account. Depression, panic disorder and social phobia showed no significant associations with platelet MAO activity. Subjects with a history of serious attempts at suicide had low platelet MAO activity; but although the difference from controls was as great as the reduction associated with smoking it was not significant after correction for smoking effects. CONCLUSIONS: Although synaptic MAO activity undoubtedly plays a role in psychopathology, the concept that platelet MAO activity is a direct genetic marker of vulnerability to alcohol dependence cannot be sustained. PMID- 10824665 TI - The well-being questionnaire: evidence for a three-factor structure with 12 items (W-BQ12). AB - BACKGROUND: The Well-being Questionnaire (W-BQ) has been designed to measure psychological well-being in people with a chronic somatic illness and is recommended by the World Health Organization for widespread use. However, studies into the factor structure of this instrument are still limited and their findings are inconsistent. This study aimed to investigate the factor structure of the Dutch version of the W-BQ. METHODS: A cross-validation design was used. A total of 1472 people with diabetes completed the W-BQ and were randomly assigned to group A or B. In group A (N = 736), exploratory factor analyses were conducted. Group B (N = 736) was split up into four subgroups of male or female patients with type 1 or type 2 diabetes. In these subgroups, confirmatory factor analyses were employed to test the model(s) developed in group A and the two models described in the literature (four-factor model with 22 items and a three-factor model with 12 items). RESULTS: Exploratory factor analyses yielded a three-factor model with 21 items (negative well-being, energy and positive well-being). In the subgroups of group B confirmatory factor analyses only accepted the three-factor model with 12 items. This factor solution was stable across gender, type of diabetes and level of education. CONCLUSIONS: The best description of the factor structure of the Dutch translation of the W-BQ was given by a three-factor solution with 12 items (W-BQ12), measuring positive well-being (four items), negative well-being (four items) and energy (four items). PMID- 10824666 TI - Binge eating and eating attitudes among Nazi concentration camp survivors. AB - BACKGROUND: Prisoners in Nazi concentration camps lived through extreme situations that included starvation. We test our hypothesis that there is a greater lifetime presence of binge eating among survivors from concentration camps than in a control group. METHODS: The subjects were 51 political prisoners who survived Nazi concentration camps and 47 ex-partisans of similar age and sex. A clinical interview investigated the lifetime occurrence of binge eating. The Eating Attitudes Test was also administered. RESULTS: The mean reported loss of weight among survivors was 27-3 kg. Thirty-three per cent of them and 4% of the ex-partisans reported going on eating binges at some time in their lives (P < 0.0007). There was no significant difference in the Eating Attitudes Test scores of survivors and ex-partisans, but, among survivors, the Bulimia subscale significantly discriminated subjects who reported current binge eating. CONCLUSIONS: Our study confirms that subjects who have survived a period of extreme food deprivation are more likely to develop binge eating behaviour. PMID- 10824667 TI - Cognitive vulnerability in patients with bipolar disorder. AB - BACKGROUND: No study has simultaneously explored key components of Beck's model of cognitive vulnerability to depression in people with bipolar disorders. METHODS: We compared 41 euthymic bipolar patients with 20 healthy control subjects. All subjects were assessed on the Hamilton Rating Scale for Depression, the Autobiographical Memory Test and the Mean Ends Problem-Solving procedure and also completed the Beck Depression Inventory, the Dysfunctional Attitude Scale, the Sociotropy Autonomy Scale and the Rosenberg Self-Esteem Questionnaire. RESULTS: In comparison to control subjects, patients with bipolar disorder demonstrated significantly higher levels of dysfunctional attitudes (particularly perfectionism and need for approval) and sociotropy, significantly greater over general recall on an autobiographical memory test and significantly less ability to generate solutions to social problem-solving tasks. These between group differences remained significant when age, intelligence, latency to respond to autobiographical memory test cue words, and subjective mood ratings were included as co-variates in the statistical analysis. Within the patient group, cognitive dysfunction was significantly correlated with level of morbidity (as measured by number of previous illness episodes). CONCLUSIONS: This study suggests that cognitive vulnerability in patients with bipolar disorder is similar to that described in unipolar disorders. It is not clear whether this dysfunction is a cause or an effect of repeated episodes of bipolar disorder. However, the findings may have implications for clinical treatment as well as suggesting a number of important new avenues of research into psychological models of affective disorder. PMID- 10824668 TI - Duration of illness and structure of symptoms in schizophrenia. PMID- 10824669 TI - The structure of genetic and environmental risk factors for three measures of disordered eating. PMID- 10824670 TI - Differential induction of gene expression by basic fibroblast growth factor and neuroD in cultured retinal pigment epithelial cells. AB - Embryonic chick retinal pigment epithelial (RPE) cells can undergo transdifferentiation upon appropriate stimulation. For example, basic fibroblast growth factor (bFGF) induces intact RPE tissue younger than embryonic day 4.5 (E4.5) to transdifferentiate into a neural retina. NeuroD, a gene encoding a basic helix-loop-helix transcription factor, triggers de novo production of cells that resemble young photoreceptor cells morphologically and express general neuron markers (HNK-1/N-CAM and MAP2) and a photoreceptor-specific marker (visinin) from cell cultures of dissociated E6 RPE (Yan & Wang, 1998). The present study examined whether bFGF will lead to the same transdifferentiation phenomenon as neuroD when applied to dissociated, cultured E6 RPE cells, and whether interplay exists between the two factors under the culture conditions. Dissociated E6 RPE cells were cultured in the presence or absence of bFGF, and with or without the addition of retrovirus expressing neuroD. Gene expression was analyzed with immunocytochemistry and in situ hybridization. Unlike neuroD, bFGF did not induce the expression of visinin, or HNK-1/N-CAM and MAP2. However, bFGF elicited the expression of RA4 immunogenicity; yet, many of these RA4-positive cells lacked a neuronal morphology. Addition of bFGF to neuroD-expressing cultures did not alter the number of visinin-expressing cells; misexpression of neuroD in bFGF-treated cultures did not change the number of RA4-positive cells, suggesting the absence of interference or synergistic interaction between the two factors. Our data indicated that bFGF and neuroD induced the expression of different genes in cultured RPE cells. PMID- 10824671 TI - Effects of atropine on refractive development, dopamine release, and slow retinal potentials in the chick. AB - Atropine has previously been found to suppress visually induced myopia both in animals and humans. The mechanism of its action is unclear. We have studied its retinal effects in an in vitro preparation, using the retina-pigment epithelium choroid complex of the chick eye. In vivo, deprivation myopia was induced by translucent goggles. Atropine solution was injected into the vitreous at two-day intervals. Dopamine release from the retina following atropine injection in vivo and from the in vitro retina preparation was quantified by HPLC-EC. In vitro preparations of the isolated chick retina-pigment epithelium-choroid were superfused with atropine. Light-induced potentials (local ERG), slow standing potentials from the retinal pigment epithelium/neural retina, and extracellular potassium concentrations were recorded. In line with previous findings, intravitreal injections of atropine (25 microg, 250 microg) reduced deprivation myopia in a dose-dependent manner. Atropine increased the release of the neurotransmitter dopamine into the superfusate in vitro at 100-500 microM and into the vitreous in vivo at 250 microg. Before an increase was measured in the vitreous, the retinal dopamine content was elevated. In concentrations equivalent to the intravitreal concentration to suppress myopia in vivo (200-800 microM), atropine induced spreading depression (SD) in the in vitro preparation. In contrast, muscarinic agonists, acetylcholine and pilocarpine, did not induce SD. Atropine reduced the ERG b- and d-wave, led to damped oscillations of RPE potentials, and reversed the ERG c-wave. Atropine suppressed myopia only at doses at which severe nonspecific side effects were observed in the retina. Atropine seems to intrude massively into the vital functions of the retina as indicated by the occurrence of SD. We conclude that atropine, by inducing SD, boosts neurotransmitter release from cellular stores, which may cancel out a presumed retinal signal that controls eye growth and through this, myopia. PMID- 10824672 TI - D2-dopamine receptor blockade impairs motion detection in goldfish. AB - Under photopic illumination conditions, motion detection in goldfish is dominated by the long-wavelength-sensitive cone type (L-cone), and under scotopic conditions motion it is determined by rods (Schaerer & Neumeyer, 1996). The switch from rod-dominated to cone-dominated motion detection occurs during light adaptation. It has been suggested that dopamine acts as a neuronal light adaptative signal. It is known that dopamine affects wavelength discrimination through D1-dopamine receptors (Mora-Ferrer & Neumeyer, 1996), and the dorsal light reflex through D1- and D2-dopamine receptors (Lin & Yazulla, 1994a). The purpose of this study was to determine whether dopamine influenced movement detection by goldfish, and if so, which dopamine receptor was involved. The D2 dopamine receptor antagonist sulpiride reduced the animal's sensitivity to the moving stimulus, whereas SCH 23390, a D1-dopamine receptor antagonist, did not have any effect. The effect of sulpiride is discussed in relation to known sulpiride effects on retinal neurons and the retinal pigment epithelium. PMID- 10824673 TI - NMDAR-1 staining in the lateral geniculate nucleus of normal and visually deprived cats. AB - In normal adult cats, a monoclonal antibody directed toward the NR-1 subunit of the N-methyl-D-aspartate (NMDA) receptor (Pharmingen, clone 54.1) produced dense cellular and neuropil labeling throughout all layers of the lateral geniculate nucleus (LGN) and adjacent thalamic nuclei, including the thalamic reticular, perigeniculate, medial intralaminar, and ventral lateral geniculate nuclei. Cellular staining revealed well-defined somata, and in some cases proximal dendrites. NMDAR-1 cell labeling was also evident in the LGN of early postnatal kittens, suggesting that developing LGN cells possess this receptor subunit at or before eye opening. Within the A-layers of the adult LGN, staining encompassed a wide range of soma sizes. Soma size comparisons of NMDAR-1 stained cells with those stained with an antibody directed toward a nonphosphorylated neurofilament protein (SMI-32), which selectively stains Y-relay cells (Bickford et al., 1998), or an antibody to glutamic acid decarboxylase (GAD), which stains for GABAergic interneurons, suggested that NMDA receptors are utilized by relay cells and interneurons. NMDAR-1 staining was also observed in the LGN of cats with early monocular lid suture. Although labeling was apparent in both deprived and nondeprived A-layers of LGN, the distribution of soma sizes was significantly different. In the deprived A-layers of LGN, staining was limited to small- and medium-sized cells. Cells with relatively large soma were lacking. However, cell density measurements as well as soma size comparisons with cells stained for Nissl substance suggested these differences were due to deprivation-induced cell shrinkage and not to a loss of NMDAR-1 staining in Y-cells. Taken together, these results suggest that NMDA receptors are utilized by both relay cells and interneurons in LGN and that alterations in early visual experience do not necessarily affect the expression of NMDA receptors in the LGN. PMID- 10824674 TI - Chloride efflux inhibits single calcium channel open probability in vertebrate photoreceptors: chloride imaging and cell-attached patch-clamp recordings. AB - The present study uses cell-attached patch-recording techniques to study the single-channel properties of Ca2+ channels in isolated salamander photoreceptors and investigate their sensitivity to reductions in intracellular Cl-. The results show that photoreceptor Ca2+ channels possess properties similar to L-type Ca2+ channels in other preparations, including (1) enhancement of openings by the dihydropyridine agonist, (-)BayK8644; (2) suppression by a dihydropyridine antagonist, nisoldipine; (3) single-channel conductance of 22 pS with 82 mM Ba2+ as the charge carrier; (4) mean open probability of 0.1; (5) open-time distribution fit with a single exponential (tau0 = 1.1 ms) consistent with a single open state; and (6) closed time distribution fit with two exponentials (tau(c1) = 0.7 ms, tau(c2) = 25.4 ms) consistent with at least two closed states. Using a Cl- -sensitive dye to measure intracellular [Cl-], it was found that perfusion with gluconate-containing, low Cl- medium depleted intracellular [Cl-]. It was therefore possible to reduce intracellular [Cl-] by perfusion with a low Cl- solution while maintaining the extracellular channel surface in high Cl- pipette solution. Under these conditions, the single-channel conductance was unchanged, but the mean open probability fell to 0.03. This reduction can account for the 66% reduction in whole-cell Ca2+ currents produced by perfusion with low Cl- solutions. Examination of the open and closed time distributions suggests that the reduction in open probability arises from increases in closed-state dwell times. Changes in intracellular [Cl-] may thus modulate photoreceptor Ca2+ channels. PMID- 10824675 TI - Response variability and information transfer in directional neurons of the mammalian horizontal optokinetic system. AB - This study is concerned with how information about the direction of visual motion is encoded by motion-sensitive neurons. Motion-sensitive neurons are usually studied using stimuli unchanging in speed and direction over several seconds. Recently, it has been suggested that neuronal responses to more naturalistic stimuli cannot be understood on the basis of experiments with constant-motion stimuli (de Ruyter van Steveninck et al., 1997). We measured the variability and information content of spike trains recorded from directional neurons in the nucleus of the optic tract (NOT) of the wallaby, Macropus eugenii, in response to constant and time-varying motion. While the NOT forms part of the mammalian optokinetic system, we have shown previously that the responses of its directional neurons resemble those of insect H1 in many respects (Ibbotson et al., 1994). We find that directional neurons in the wallaby NOT respond with lower variability and higher rates of information transmission to time-varying stimuli than to constant motion. The difference in response variability is predicted by an inhomogeneous Poisson model of neuronal spiking incorporating an absolute refractory period of 2 ms during which no subsequent spike can be fired. Refractoriness imposes structure on the spike train, reducing variability (de Ruyter van Steveninck & Bialek, 1988; Berry & Meister, 1998). A given refractory period has a greater impact when firing rates are high, as for the responses of NOT neurons to time-varying stimuli. It is in just these cases that variability in experimentally observed spike trains is lowest. Thus, differences in response variability do not necessarily imply that different models are required to predict neuronal responses to constant- and time-varying motion stimuli. PMID- 10824676 TI - Regulation of arrestin mRNA levels in Limulus lateral eye: separate and combined influences of circadian efferent input and light. AB - Most animals experience daily changes in light and darkness. The retinas of many of these animals show concomitant rhythmic changes in the levels of mRNAs that encode proteins involved in the photoresponse. These changes may be circadian and independent of light, independent of circadian clocks and regulated by light, or regulated by a circadian clock and light. We have taken advantage of the organization of the Limulus visual system to examine the separate and combined effects of signals from a circadian clock and light on arrestin mRNA levels in photoreceptors. The clock that regulates photoreceptors in the lateral eye of Limulus is in the brain, and signals from the clock reach the lateral eye via activation of a well-characterized efferent projection in the lateral optic nerve. In the experiments described, clock-driven efferent input to the lateral eye was eliminated by cutting the lateral optic nerve, and light input to the lateral eye was eliminated by placing an opaque patch over the eye. Arrestin mRNA levels were quantified relative to 18s rRNA with a ribonuclease protection assay. We observed the following. In lateral eyes exposed to natural diurnal light and endogenous efferent nerve activity, the level of arrestin mRNA was higher during the day in the light than during the night in the dark. Circadian efferent nerve activity was necessary and sufficient to produce normal daily fluctuations in the level of arrestin mRNA. Light influenced arrestin mRNA levels only in eyes with intact and active efferent projections. We conclude that arrestin mRNA levels in lateral eye photoreceptors are controlled entirely by efferent nerve activity, and that light exerts its effects by modulating this output from the circadian clock. Light-stimulated changes in arrestin mRNA in the vertebrate retina may likewise require interactions between light-driven biochemical cascades and clock output. PMID- 10824677 TI - Spontaneous activity in developing turtle retinal ganglion cells: statistical analysis. AB - We report on the temporal properties of the spontaneous bursts of activity in the developing turtle retina. Quantitative statistical criteria were used to detect, cluster, and analyze the temporal properties of the bursts. The interburst interval, duration, firing rate, and number of spikes per burst varied widely among cells and from burst to burst in a single cell. Part of this variability was due to the positive correlation between a burst's duration and the interburst interval preceding that burst. This correlation indicated the influence of a refractory period on the bursts' properties. Further evidence of such a refractoriness came from the bursts' auto-covariance function, which gives the tendency of a spike to occur a given amount of time after another spike. This function showed a positive phase (between approximately 10 ms and 10 s) followed by a negative one (between 10 s and more than 100 s), suggestive of burst refractoriness. The bursts seemed to be propagating from cell to cell, because there was a long (symmetrically distributed) delay between the activation of two neighbor cells (median absolute delay = 2.3 s). However, the activity often failed to propagate from one cell to the other (median safety factor = 0.59). The number of spikes per burst in neighbor cells was statistically positively correlated, indicating that the activity in the two cells was driven by the same excitatory process. At least two factors contribute to the excitability during bursts, because the positive phase of the cross-covariance function (similar to auto-covariance but for two cells) had a temporally asymmetric fast component (1 3 ms) followed by a temporally symmetric slow component (1 ms to 10 s). PMID- 10824678 TI - Inhibition is not required for the production of transient spiking responses from retinal ganglion cells. AB - Ganglion cells responding only transiently to changes in illumination are found in many different vertebrate retinas. The interactions underlying formation of these transient responses are still poorly understood. Two recently proposed hypotheses are (1) functional inhibitory pathways are necessary for transient response production, and (2) direct inhibition of the ganglion cell has little effect on its output. Here, we examine these conclusions by using cell-attached patch-clamp recordings of spiking, whole-cell recordings of synaptic currents, and computer modeling. We found that picrotoxin (a GABA(A) and GABA(C) receptor antagonist), bicuculline (a GABA(A) receptor antagonist), and strychnine (a glycine receptor antagonist), applied either singly or in combination, always failed to convert transient responses to sustained responses. Application of the GABA(B) antagonist CGP35348 in the presence of picrotoxin and strychnine also failed to convert transient responses into sustained responses. Whole-cell recordings of synaptic currents at various holding potentials indicated that direct inhibitory inputs to ganglion cells limit the duration of net excitation, implying that direct inhibition does act to truncate the ganglion cell spiking response. Computer simulations using spiking and synaptic data from combined cell attached and whole-cell recordings supported this interpretation. We conclude that inhibitory pathways are not required for generation of transient responses, but these pathways do serve to modulate transient ganglion cell spiking responses. We find that this modulation occurs, in part, via inhibitory inputs directly to the ganglion cell. PMID- 10824679 TI - Dye coupling in horizontal cells of developing rabbit retina. AB - In the mature rabbit retina, two classes of horizontal cells, A type and B type, provide lateral inhibition in the outer plexiform layer (OPL) and spatially modify the activation of bipolar cells by photoreceptors. Gap junctions connecting homologous horizontal cells determine the extent to which this inhibitory activity spreads laterally across the OPL. Little is currently known about the expression of gap junctions in horizontal cells during postnatal development or how cell-cell coupling might contribute to subsequent maturational events. We have examined the morphological attributes and coupling properties of developing A and B type horizontal cells in neonatal rabbit retina using intracellular injections of Lucifer Yellow and Neurobiotin. Prelabeling with DAPI permitted the targeting of horizontal cell bodies for intracellular injection in perfused preparations of isolated retina. A and B type horizontal cells were identifiable at birth although their dendritic field sizes had not reached adult proportions and their synaptic contacts in the OPL were minimal. Both cell types exhibited homologous dye coupling at birth. Similar to that seen in the adult, no heterologous coupling was observed, and homologous coupling among A type cells was stronger than that observed among B type cells. The spread of tracer compounds through gap junctions of morphologically immature horizontal cells suggests that ions and other small, bioactive compounds may likewise spread through coupled, horizontal networks to coordinate the subsequent maturational of emerging outer plexiform layer pathways. PMID- 10824680 TI - Computation of motion direction by quail retinal ganglion cells that have a nonconcentric receptive field. AB - One type of retinal ganglion cells prefers object motion in a particular direction. Neuronal mechanisms for the computation of motion direction are still unknown. We quantitatively mapped excitatory and inhibitory regions of receptive fields for directionally selective retinal ganglion cells in the Japanese quail, and found that the inhibitory regions are displaced about 1-3 deg toward the side where the null sweep starts, relative to the excitatory regions. Directional selectivity thus results from delayed transient suppression exerted by the nonconcentrically arranged inhibitory regions, and not by local directional inhibition as hypothesized by Barlow and Levick (1965). PMID- 10824681 TI - Modulation by Zn2+ of GABA responses in bipolar cells of the mouse retina. AB - The localization of endogenous Zn2+ in the mouse retina was examined histochemically and the inhibitory action of Zn2+ on GABA-induced responses was studied in bipolar cells isolated from the mouse retina. Accumulation of endogenous Zn2+ was detected in photoreceptors, bipolar, and/or amacrine cells by either the bromopyridylazo-diethylaminophenol method or the dithizone method. Under whole-cell recording conditions, GABA induced a Cl- current in isolated bipolar cells. The current consisted of two components. The first component was inhibited completely by application of 100 microM bicuculline, suggesting that this is a GABA(A)-receptor mediated current. The second component was inhibited completely by 100 microM 3-aminopropyl-(methyl)-phosphinic acid, suggesting that this is a GABA(C)-receptor mediated current. GABA(C) receptors were present at a higher density on the axon terminal than on dendrites. Zn2+ inhibited both GABA(A) and GABA(C) receptors. GABA(C) receptors were more susceptible to Zn2+; the IC50 for the GABA(A) receptor was 67.4 microM and that for the GABA(C) receptor was 1.9 microM. These results suggest that Zn2+ modulates the inhibitory interaction between amacrine and bipolar cells, particularly that mediated by the GABA(C) receptor. PMID- 10824683 TI - The role of NMDA channels in rabbit retinal directional selectivity. AB - It has been previously demonstrated that the majority of the glutamatergic input to directionally selective (DS) ganglion cells in the rabbit retina is mediated by NMDA receptors. To examine whether NMDA channels have any role in directional selectivity, we eliminated magnesium from the superfusion medium to prevent the magnesium block of the channels at hyperpolarized membrane potentials. During superfusion in magnesium-free media, the response to null-direction motion increased to the level of the response to preferred-direction motion. This effect was specifically mediated by NMDA channels because subsequent blocking of the NMDA channels with AP7 restored directional selectivity. We also tested whether the increase in the null-direction response in magnesium-free medium was due to an increased release of acetylcholine from the cholinergic amacrine cells, rather than an effect on the DS ganglion cells themselves, by blocking acetylcholine transmission with d-tubocurarine during superfusion with the magnesium-free medium. During zero-magnesium superfusion, d-tubocurarine reduced both the preferred- and null-direction responses of DS ganglion cells but did not restore directional selectivity. These findings suggest that null-direction motion normally causes portions of the dendritic membrane of the directionally selective ganglion cell to be maintained at a sufficiently negative potential that the NMDA channels are blocked by magnesium ions. This result is discussed in terms of several models for the mechanisms of directional selectivity. PMID- 10824682 TI - The functional influence of nicotinic cholinergic receptors on the visual responses of neurones in the superficial superior colliculus. AB - In the rat, the superficial gray layer (SGS) of the superior colliculus receives glutamatergic projections from the contralateral retina and from the visual cortex. A few fibers from the ipsilateral retina also directly innervate the SGS, but most of the ipsilateral visual input is provided by cholinergic afferents from the opposing parabigeminal nucleus (PBG). Thus, visual input carried by cholinergic afferents may have a functional influence on the responses of SGS neurones. When single neuronal extracellular recording and iontophoretic drug application were employed to examine this possibility, cholinergic agonists were found to depress responses to visual stimulation. Lobeline and 1-acetyl-4 methylpiperazine both depressed visually evoked activity and had a tendency to reduce the background firing rate of the neurones. Carbachol depressed the visual responses without any significant effect on the ongoing activity, while the muscarinic receptor selective agonist methacholine increased the background activity of the neurones and reduced their visual responses. Lobeline was chosen for further studies on the role of nicotinic receptors in SGS. Given that nicotinic receptors are associated with retinal terminals in SGS, and that the activation of presynaptic nicotinic receptors normally facilitates transmitter release (in this case glutamate release), the depressant effects of nicotinic agonists are intriguing. However, many retinal afferents contact inhibitory neurones in SGS; thus it is possible that the increase in glutamate release in turn facilitates the liberation of GABA which goes on to inhibit the visual responses. We therefore attempted to reverse the effects of lobeline with GABA receptor antagonists. The depressant effects of lobeline on the visual response could not be reversed by the GABA(A) antagonist bicuculline, but the GABA(B) antagonist CGP 35348 reduced the effects of lobeline. We hypothesize that cholinergic drive from the parabigeminal nucleus may activate presynaptic nicotinic receptors on retinal terminals, thereby facilitating the release of glutamate onto inhibitory neurones. Consequently GABA is released, activating GABA(B) receptors, and thus the ultimate effect of nicotinic receptor activation is to depress visual responses. PMID- 10824684 TI - Cloning and tissue localization of a novel zebrafish RdgB homolog that lacks a phospholipid transfer domain. AB - The retinal degeneration B (RdgB) protein family is characterized by an amino terminal phosphatidylinositol transfer protein (PITP) domain, several hydrophobic domains, and a highly conserved carboxyl terminus. We identified a zebrafish RdgB homolog (pl-RdgB) that lacks the amino-terminal PITP domain, while retaining over 45% amino acid identity with the two mouse RdgB proteins (M-RdgB1 and M-RdgB2). Unlike the widespread retinal expression observed for other vertebrate RdgB homologs, pl-RdgB is restricted in the retina to the cone cell inner segments. The pl-RdgB protein is also expressed in the brain, although its distribution is different than the other RdgB homologs. Analogous to M-RdgB2, pl-RdgB protein is extracted from a retinal homogenate by guanidine and not by Triton X-100. Thus, pl-RdgB and likely all the identified RdgB homologs are not integral membrane proteins, but may associate with the membrane through protein-protein interactions. While expression of either murine RdgB homolog restored the defective light response and prevented retinal degeneration in rdgB mutant flies, expressing zebrafish pl-RdgB in Drosophila rdgB2 null mutants slowed retinal degeneration without restoring the electrophysiological light response. Thus, pl RdgB may define a previously unrecognized protein family, which includes the other RdgB homologs, that act through a protein complex to maintain photoreceptor viability. PMID- 10824685 TI - Organization of brain stem afferents to the ventral lateral geniculate nucleus of rats. AB - We have examined the patterns of projections from different nuclei of the brain stem to the ventral lateral geniculate nucleus (vLGN) of the thalamus. Injections of biotinylated dextran were made into different nuclei of the brainstem (i.e., midbrain reticular nucleus, pontine reticular nucleus, deep layers of superior colliculus, periaqueductal grey matter [ventrolateral, dorsolateral, and lateral columns], pedunculopontine tegmental nucleus, parabrachial nucleus, lateral dorsal tegmental nucleus, substantia nigra [pars reticulata], locus coeruleus, and dorsal raphe) of Sprague-Dawley rats using stereotaxic coordinates. Our results show that all of the above mentioned brain-stem nuclei have overlapping projections to the medial regions of the vLGN, within the parvocellular lamina of the nucleus. This if the first instance of the parvocellular lamina being shown to receive a major set of projections. Very few labelled terminals from the brain stem were ever seen within the larger more lateral magnocellular lamina, which has been shown by previous studies to receive heavy inputs from visually associated structures, such as the retina and occipital cortex. Since many of the brain-stem nuclei injected in this study have little to do with visual processing, our results suggest that one can perhaps package the vLGN into distinct visual (magnocellular) and nonvisual (parvocellular) components. PMID- 10824686 TI - Subcellular distribution and characterization of rat pancreatic phospholipase D isoforms. AB - This study was undertaken to characterize the biochemical properties of rat pancreatic phospholipase D (PLD). Based on Western blot analysis of pancreas subcellular fractions, PLD1 was detected as a protein of 120 kDa associated with the microsomal fraction, whereas PLD2 appeared as a 105-kDa protein enriched in the microvesicular fraction. In these fractions, a low level of PLD activity was measured with an exogenous substrate containing phosphatidylinositol-4,5 bisphosphate (PIP2), unresponsive to guanosine triphosphate (GTP)gammaS and adenosine diphosphate (ADP)-ribosylation factor (ARF). Addition of unsaturated but not saturated fatty acids stimulated an oleate-dependent PLD activity that colocalized with the PLD1 enzyme in the crude plasma membrane and microsomal fractions. The transphosphatidylation reaction was maximal with either 200-400 mM (1.2-2.3%) ethanol or 25 mM (0.23%) 1-butanol, with an optimal pH between 6.5 and 7.2. Lipids extracted from the pancreatic membranes were potent inhibitors of the HL60 cell PLD activity when compared with those isolated from HL60 cells. Oleate dependent PLD activity was less susceptible to these inhibitions. A phospholipase A1 (PLA1) activity hydrolyzing phosphatidylethanol also was found in the pancreatic membrane fractions and was nearly absent in the HL60 cells. This activity was completely inhibited by 400 nM tetrahydrolipstatin (THL), a lipase inhibitor. Pancreatic PLD1 and PLD2 activities could be measured after a chromatographic separation from microsomal membranes and high-speed supernatants, respectively. Activities of both enzymes were inhibited by oleate and required the presence of PIP2 in the substrate vesicles. ARF1 strongly activated PLD1 in a dose-dependent manner, and PLD2 was slightly responsive. Indirect immunofluorescence revealed that PLD2 is distributed throughout the pancreas, with a more intense staining in the islets. This study presents for the first time biochemical characteristics of the pancreatic PLD activities and shows the presence of oleate-dependent PLD1 and PLD2 activities, as well as PLD1 and PLD2 proteins in this gland. PMID- 10824687 TI - Differentiation of islet cells in long-term culture. AB - Our previous studies in the hamster pancreatic cancer model have shown that exocrine pancreatic cancer arises from ductal/ductular cells, as well as from within the islets, most probably from islet precursor (stem) cells. To identify and characterize these cells, we established a long-term culture from isolated hamster islets and investigated their growth, differentiation, and expression of biomarkers. Islets maintained their original form and structure within the first 14 days in culture. However, beginning at day 7, ductular structures began to form within the islets. At day 21 in culture, acinar cells, intermediary cells, oncocytes, and cells comparable to pancreatic hepatocytes also appeared between ductular and endocrine cells. The number of duct-like cells gradually increased, whereas the number of hormone-producing cells decreased. After 35 days in culture, the exocrine cells disappeared, and undifferentiated cells formed a monolayer. These cells expressed cytokeratins, alpha1-antitrypsin, transforming growth factor-alpha, epidermal growth factor receptor, carbonic anhydrase II, vimentin, laminin, and showed binding to tomato lectin and Phaseolus vulgaris leukoagglutinin. They did not express the regulatory transcriptional factors, insulin-promoting factor 1, NKx6.1, Pax6, and NeuroD. The results thus indicate that islet cells have potential to form exocrine cells. At present, it is not clear whether these cells originate from preexisting stem cells or from transdifferentiated islet cells. PMID- 10824688 TI - Oxyntomodulin inhibits pancreatic secretion through the nervous system in rats. AB - Glicentin (GLIC), oxyntomodulin (OXM), and peptide YY (PYY) released in blood by ileocolonic L-cells after meals may inhibit pancreatic secretion. Whereas OXM interacts with glucagon and tGLP-1 receptors, OXM 19-37, a biologically active fragment, does not. The purpose of this study was to measure the effect of OXM, OXM 19-37, GLIC, tGLP-1, and PYY on pancreatic secretion stimulated by 2 deoxyglucose (2DG), electrical stimulation of the vagus nerves (VES), acetylcholine and cholecystokinin octapeptide (CCK8) in anesthetized rats. The effect of OXM was also studied in dispersed pancreatic acini. Plasma oxyntomodulin-like immunoreactivity (OLI) was measured by radioimmunoassay after the exogenous infusion of OXM and after an intraduodenal meal. OXM 19-37, infused at doses mimicking postprandial plasma levels of OLI, decreased pancreatic secretion stimulated by 2DG, VES, or CCK8. Similar effects were found with OXM and GLIC. OXM 19-37 did not change the pancreatic stimulation induced by acetylcholine in vivo, or CCK-induced amylase release in isolated acini. Vagotomy completely suppressed the inhibitory effect of OXM 19-37 on CCK8-stimulated pancreatic secretion. PYY inhibited the effect of 2DG, but not that of CCK8, whereas tGLP-1, even in pharmacologic doses, had no effect on stimulated pancreatic secretion. OXM, OXM 19-37, but not tGLP-1, inhibit pancreatic secretion at physiologic doses, through a vagal neural indirect mechanism, different from that used by PYY, and probably through a GLIC-related peptide specific receptor. PMID- 10824689 TI - Autonomic nervous function in alcohol-related chronic pancreatitis. AB - Patients with alcohol-related chronic pancreatitis (ARCP) often have peripheral neuropathy, but no data on the occurrence of autonomic neuropathy (AN) are available for this condition. To assess the autonomic function and the significance of its abnormalities for the prognosis of ARCP, 18 patients with ARCP and associated diabetes mellitus (P-DM group), 10 with ARCP without evidence of diabetes mellitus (P group), 17 patients with insulin-dependent diabetes mellitus (IDDM group), and 18 healthy controls answered a structured questionnaire and underwent three standardized cardiovascular (CV) tests that yielded six different parameters for autonomic nerve function. Patients with at least one symptom plus two abnormal results on CV tests were regarded as having AN. ARCP patients were followed up for 48 months and mortality rates were recorded. The proportions of patients with AN were 66.6% in the P-DM group, 30.0% in the P group, and 29.4% in IDDM patients. Seven of 15 ARCP patients with AN died during follow-up, compared with one of 13 of those without AN (p < 0.037). In conclusion, AN is commonly found in ARCP patients and carries an ominous prognosis. PMID- 10824690 TI - Hemoconcentration is an early marker for organ failure and necrotizing pancreatitis. AB - In a previous retrospective case-control study, hemoconcentration was associated with the development of pancreatic necrosis. The aim of the present study was to determine in a cohort study whether hemoconcentration is a marker for both organ failure and necrotizing pancreatitis. A cohort study was performed on patients admitted with acute pancreatitis from February 1996 to April 1997. Pancreatic necrosis was defined by findings on dynamic contrast-enhanced computed tomography scan or magnetic resonance imaging. Of 128 total patients with acute pancreatitis, 53 underwent computed tomography or magnetic resonance imaging. Eighteen of 53 had necrotizing pancreatitis. Logistic regression identified an admission hematocrit > or = 44% and a failure of admission hematocrit to decrease at 24 hours as the best binary predictors of necrotizing pancreatitis and organ failure. By 24 hours, 17 of 18 patients with necrotizing pancreatitis versus 11 of 35 with interstitial pancreatitis met one or the other criterion for necrosis (p < 0.001). By 24 hours, 13 of 15 with organ failure versus 36 of 104 without organ failure met one or the other criterion (p < 0.001). The negative predictive value by 24 hours was 96% for necrotizing pancreatitis and 97% for organ failure. Hemoconcentration with an admission hematocrit > or = 44% and/or failure of admission hematocrit to decrease at approximately 24 hours was associated with the development of necrotizing pancreatitis and organ failure. Patients who did not experience hemoconcentration were very unlikely to develop pancreatic necrosis or organ failure. PMID- 10824691 TI - Therapeutic effect of basic fibroblast growth factor on experimental pancreatitis in rat. AB - Basic fibroblast growth factor (bFGF) is one of the mitogens that facilitate endothelial proliferation and angiogenesis. This study was designed to examine the therapeutic effect of bFGF on experimental pancreatitis in rat. Edematous pancreatitis was induced by intraperitoneal injections of cerulein (50 microg/kg) at hourly intervals. BFGF (70 nmol/kg) was administered intraperitoneally after induction of pancreatitis. DNA synthesis of isolated pancreatic acinar cells of normal rats was determined as the uptake of 5-bromo-2'-deoxyuridine (BrdU) into the cells. Immunohistochemical staining of DNA synthesis in acinar cells during cerulein-induced pancreatitis was also examined with BrdU labeling in vivo technique. Cerulein administration increased serum amylase, lipase level, and wet weight of pancreatic tissue. Treatment with bFGF markedly ameliorated all these parameters. In primary culture system of isolated pancreatic acinar cells of normal rats, bFGF caused a dose-dependent increase in BrdU incorporation into DNA, showing an EC50 value of 0.8 nmol/L and a maximum response of 2.5-fold increase at a concentration of 400 nmol/L. bFGF treatment (70 nmol/kg) markedly increased BrdU labeling in the nucleus of acinar cells of the pancreatitis rats group in immunohistochemical examination when compared with control without bFGF treatment. Treatment with bFGF may represent a promising therapeutic concept for patients with acute pancreatitis. PMID- 10824692 TI - The usefulness of serial changes in serum CA19-9 levels in the diagnosis of pancreatic cancer. AB - We examined serial changes in serum CA19-9 levels in patients with pancreatic cancer (PC) or chronic pancreatitis (CP) to clarify whether these changes are useful in the diagnosis of PC. We chose II PC patients in whom serum CA19-9 levels had been measured serially before a definitive diagnosis of PC had been established. We also examined serial changes in serum CA19-9 levels during the recent 1-year follow-up period in 21 CP patients. Serial changes in these levels were classified into the following five types; normal, elevated, increased, decreased, and other. Of the 11 PC patients, seven patients (64%) showed the increased type, whereas none showed the normal type. However, among the CP patients, 10 patients (48%) showed the normal type and only one (5%) showed the increased type. The proportion of the increased type among PC patients was significantly higher than that among CP patients. In conclusion, serial changes in serum CA19-9 levels may be useful to differentiate between PC and CP. Close follow-up with various diagnostic modalities may be required for patients whose serum CA19-9 levels increase to more than twofold the initial level, because such findings are highly suggestive of PC. PMID- 10824693 TI - Elevated serum levels of antibodies to carbonic anhydrase I and II in patients with chronic pancreatitis. AB - An immune-mediated reaction to pancreatic structures has been postulated for the pathogenesis of chronic pancreatitis (CP). Several reports demonstrate the presence of antibodies to the pancreatic ductal epithelium in some patients suffering from CP. Serum antibodies to carbonic anhydrase I (anti-CA I) and II (anti-CA II) are present in patients affected by idiopathic CP. The aim of this study was to evaluate the presence of anti-CA I and anti-CA II in a series of patients with CP. We studied 78 consecutive CP patients (62 male, 16 female; mean age 48.6 +/- 10.2 years) referred to the Verona University Center for the Study of the Pancreas. As a control group, we studied 26 healthy subjects recruited from among the medical and nursing staff of the center. Serum anti-CA I and anti CA II levels were quantified by enzyme-linked immunosorbent assay using a standard method with minor modifications. The mean absorbance of antibodies was higher in CP patients than in control subjects (anti-CA I: 0.064 +/- 0.042 vs. 0.047 +/- 0.015, p = 0.051; and anti-CA II: 0.038 +/- 0.02 vs. 0.029 +/- 0.014, p = 0.033). Positive results were arbitrarily defined as absorbance values >0.067 for anti-CA I and 0.047 for anti-CA II. We found anti-CA I and anti-CA II positivity in 21 of 78 (27%) and 20 of 78 (26%) of CP patients, respectively, and in only two of 26 control subjects (7.7%) (p = 0.032 and 0.039). Twenty-two of 26 subjects in the control group (84.6%) and 48 of 78 patients (61.5%) in the CP group tested negative for both antibodies (p = 0.03). None of the control subjects and 12 of 78 (16.6%) of the CP patients tested positive for both anti-CA I and anti-CA II. We observed a significant correlation between anti-CA I and anti-CA II serum levels in control subjects (R = 0.423; p = 0.016) and in CP patients (R = 0.584; p < 0.0001). No correlation was found between serum antibody levels and any of the following variables: length of disease, alcohol consumption, smoking habits, pancreatic surgery, pancreatic calcifications, diabetes, and steatorrhea. Serum levels of anti-CA I and anti-CA II are elevated in some patients suffering from CP. PMID- 10824694 TI - Do plasma and urine trypsinogen activation peptides (TAP) really increase in trypsin-taurocholate-induced pancreatitis? AB - Plasma and urine levels of trypsinogen activation peptides (TAP) reflect the severity of acute pancreatitis in experimental and clinical acute pancreatitis. In trypsin-taurocholate-induced pancreatitis in rats, the extrinsic bovine trypsin used for the induction of pancreatitis might influence on the TAP levels after induction of pancreatitis. The aim of the present study was to elucidate whether infused trypsin itself affects TAP levels in trypsin-taurocholate-induced pancreatitis. Rats were divided into three groups. In the pancreatitis group, acute pancreatitis was induced by a retrograde infusion of bovine trypsin and sodium taurocholate into the pancreatic duct. In the duct infusion group and peritoneal injection group, a mixture of bovine trypsin and trypsin inhibitor, ONO-3403, was infused into the pancreatic duct or the peritoneal cavity. Plasma and urine TAP concentration significantly increased in trypsin-taurocholate induced pancreatitis but not in the duct infusion and peritoneal injection groups for 6 hours after the infusion of trypsin. Serum rat immunoreactive trypsin (IRT) and amylase significantly increased in the pancreatitis and duct infusion groups but not in the peritoneal injection group. Serum levels of bovine IRT in the pancreatitis group was significantly lower than those in duct infusion and peritoneal injection groups. In conclusion, an intraductal infusion of bovine trypsin itself into pancreatic duct does not influence the levels of plasma and urine TAP in trypsin-taurocholate-induced pancreatitis. PMID- 10824695 TI - Oral administration of a synthetic trypsin inhibitor increases pancreatic duct function in CCK-A receptor-deficient rats. AB - The effects of oral administration of a synthetic trypsin inhibitor on bicarbonate secretion were examined in cholecystokinin A (CCK-A) receptor deficient (OLETF) rats and compared with Wistar rats. Rats were fed chow containing 0.1% trypsin inhibitor for 7 days. Rats were prepared with cannulae draining bile and pancreatic juice separately and with duodenal and extrajugular vein cannulae after 3-day trypsin inhibitor ingestion. Then the animals were maintained in Bollman cages, and the experiments were conducted 4 days after surgery. After 1.5 h of basal secretion with bile-pancreatic juice return, bile pancreatic juice was diverted for 2 h. The responses of bicarbonate secretion to bile-pancreatic juice diversion were significantly enhanced in rats treated with trypsin inhibitor compared with those given a control diet, whereas responses of fluid and protein secretion were not affected in OLETF rats. The response of protein secretion, but not those of fluid or bicarbonate secretion, was enhanced in Wistar rats by treatment with trypsin inhibitor. Carbonic anhydrase II gene expression was increased by 7-day treatment with trypsin inhibitor only in OLETF rats, and not in Wistar rats. PMID- 10824696 TI - Endogenous nitric oxide mediates pancreatic exocrine secretion stimulated by secretin and cholecystokinin in rats. AB - Nitric oxide (NO) is one of the important biologic mediators in regulation of gastrointestinal (GI) functions, but the influence of NO on the release of secretin and cholecystokinin (CCK) and exocrine pancreatic secretion has not been adequately investigated in the rat. The aim of this study was to determine the role of NO on endogenous and exogenous secretin- or CCK-stimulated pancreatic exocrine secretion both in anesthetized and conscious rats. Experiments were carried out in four different groups of rats with duodenal pancreatobiliary cannulas and jugular vein catheters. Group 1: During duodenal infusion of 0.05N HCl or 15% casein (pH 7.0), N-nitro-L-arginine (NNA), an inhibitor of NO-synthase in graded doses (2.5, 5, 10 mg/kg/h), was infused intravenously. Group 2: One hour after starting intravenous secretin at 5 pmol/kg/h or intravenous CCK-8 at 0.06 microg/kg/h, NNA in graded doses was administered intravenously. Group 3: In conscious rats, NNA (5 mg/kg/h) was given intravenously for 1 hour after a meal. Group 4: L-Arginine at 100 mg/kg/h was infused intravenously during the period of NNA (5 mg/kg/h) infusion in groups 1, 2, and 3. Pancreatic juice was collected at 30-minute intervals to measure volume, as well as output of bicarbonate and protein. At the end of the experiment, plasma secretin, vasoactive intestinal polypeptide (VIP) and CCK levels were determined by radioimmunoassay (RIA). NNA dose dependently inhibited the pancreatic secretion of fluid and bicarbonate stimulated by duodenal acidification, exogenous secretin, and a meal. NNA dose dependently inhibited the pancreatic secretion of protein stimulated by duodenal infusion of casein, exogenous CCK, and a meal. L-Arginine significantly reversed the NNA-induced inhibition of pancreatic secretion in all experiments. NNA did not alter significantly the plasma levels of secretin, VIP, and CCK. Our results indicated that endogenous NO plays a significant role in the regulation of pancreatic exocrine secretion stimulated by secretin and CCK. However, NO does not influence the release of secretin, VIP, or CCK in the rat. PMID- 10824697 TI - An annular pancreas derived from paired ventral pancreata, supporting Baldwin's hypothesis. AB - An annular pancreas is a rare malformation. It is generally accepted that the ring formation originates from a single ventral pancreas, as suggested by Lecco. No reports of resected and/or autopsied annular pancreata derived from paired ventral pancreata, thus supporting Baldwin's hypothesis, have been published. We describe an annular pancreas originating from paired ventral pancreata, thus supporting Baldwin's hypothesis, and attempt to clarify the pathogenesis of an annular pancreas. The patient was a 1-day-old Japanese male newborn, born at 32 weeks of pregnancy. He died from respiratory failure owing to esophageal atresia the next day. Autopsy incidentally revealed an annular pancreas that was examined histologically. Multiple 4 microm thick serial sections were obtained from paraffin-embedded pancreatic tissue. Sections for histologic analysis were subjected to hematoxylin-eosin staining and pancreatic polypeptide immunostaining. An unusually large pancreatic duct encircled by pancreatic tissue ran around the duodenum, and the duct was confirmed to flow into the major papilla. The islets of the encircling pancreas were positive for pancreatic polypeptide. A normal main and accessory pancreatic duct were also identified. Histologic and immunohistochemical evaluation revealed that the ring formation originated from the left lobe of paired ventral pancreata. An annular pancreas that was investigated histopathologically and immunohistochemically and found to support Baldwin's hypothesis is described. PMID- 10824698 TI - Evidence of protein synthesis during resuscitation of ischemically damaged canine pancreas by the two-layer method. AB - The two-layer method, which supplies sufficient oxygen to the canine pancreas graft and allows adenotriphosphate synthesis within the graft, resuscitates the ischemically damaged pancreas graft during mild hypothermic (20 degrees C) preservation, but the mechanisms are unknown. The purpose of this study was to determine whether protein synthesis was performed on ischemically damaged pancreas graft during preservation by the two-layer method in a canine autotransplantation model. The pancreas grafts subjected to 90 minutes of warm ischemia were preserved by the two-layer method (perfluorochemical/University of Wisconsin solution) at 20 degrees C for 5 hours. Graft viability was judged from graft survival after autotransplantation. DNA, RNA, and protein synthesis was quantitated by determining incorporation of tritiated thymidine, tritiated uridine, and tritiated leucine, respectively, during preservation. Significant increases in RNA and subsequent protein syntheses were observed during preservation by the two-layer method. In contrast, DNA synthesis did not follow RNA and protein synthesis. We conclude that protein is synthesized in the process of postischemic cellular recovery of the pancreas graft during mild hypothermic preservation by the two-layer method. PMID- 10824699 TI - Type 1 diabetes mellitus associated with clinical acute pancreatitis in an adult. PMID- 10824700 TI - Chronic hemosuccus pancreaticus: a rare complication of pancreatic microcystic adenoma successfully treated with Whipple's procedure. PMID- 10824701 TI - A case of small pancreatic ductal carcinoma accompanied by intraductal mucin producing pancreatic tumors. PMID- 10824702 TI - Premature trypsinogen activation during cerulein pancreatitis in rats occurs inside pancreatic acinar cells. PMID- 10824703 TI - pH modulator chloroquine prevents diet-induced acute pancreatitis. PMID- 10824704 TI - Asthma prevention and management guidelines. Ministry of Health and Welfare, Japan. PMID- 10824705 TI - Coexistence of intestinal trefoil factor (hITF) and oxytocin in magnocellular neurons in the human hypothalamus. AB - Human intestinal trefoil factor hITF, a polypeptide of the P-domain family, was found to occur in hypothalamic neurons. With combined immunofluorescence and immunoperoxidase technique we investigated the coexistence of hITF with the neurohypophysial peptide oxytocin and the associated neurophysin I in sections of the human hypothalamus. In the supraoptic nucleus, 39.2% of magnocellular oxytocinergic perikarya show hITF immunoreactivity. A similar distribution was observed in perivascular hypothalamic oxytocinergic neurons, whereas in the paraventricular nucleus, 99% of the oxytocinergic neurons show hITF coexpression. In the periventricular nucleus (PEV), single, scattered neurons with both immunoreactivities occur. Our findings indicate that hITF and oxytocin are coexpressed in a portion of the magnocellular neurons in the human hypothalamus, and that hITF is among the neurohypophysial peptides. PMID- 10824706 TI - The effect of epitestosterone on estrogen biosynthesis in vitro. AB - OBJECTIVE: The concentrations of epitestosterone in human serum correlates negatively with that of estradiol. The possible explanation of this relation was addressed, and the influence of epitestosterone on kinetics of estradiol formation in vitro was evaluated. METHODS: The concentration of epitestosterone was measured in serum of 54 men participating in a screening program for prostate disease. Epitestosterone inhibition of aromatase and 17beta-hydroxysteroid dehydrogenase activities was tested in vitro in the system consisting of human placental microsomes, NADPH or NAD and NADP respectively, and epitestosterone in increasing concentrations. Testosterone, androstenedione, estrone and 17beta estradiol were utilized as substrates. RESULTS: A significant negative correlation between epitestosterone and estradiol levels in human male serum was found. No inhibition of aromatase activity was observed; however, inhibition of 17beta-hydroxysteroid dehydrogenase was found preferentially in the direction leading to oxidation of the C-17 hydroxy group. The inhibitory effect of epitestosterone was more pronounced with androgens as substrates. CONCLUSION: Epitestosterone could influence the formation of estradiol in vitro rather by inhibition of 17beta-hydroxysteroid dehydrogenase than by blocking aromatase activity. PMID- 10824707 TI - Antihyperglycemic effects of 3-O-methyl-D-chiro-inositol and D-chiro-inositol associated with manganese in streptozotocin diabetic rats. AB - The acute effects after administration of 3-O-Methyl-D-Chiro-inositol, D-Chiro inositol and manganese, components of the pH 2.0 putative mediator of insulin action, on plasma glucose were examined in low dose streptozotocin-treated rats. 3-O-Methyl-D-Chiro-inositol at a bolus dose of 5 mg/kg decreased plasma glucose 6% at 120 min (p < 0.05). A higher bolus dose of 3-O-Methyl-D-Chiro-inositol (15 mg/kg) promoted a more persistent hypoglycemic effect of 22% at 120 min (p < 0.05). Infusion of 8.3 microg/min of manganese chloride lowered plasma glucose by 23% (p < 0.05). 3-O-Methyl-D-Chiro-inositol (15 mg/kg) together with manganese chloride (8.3 microg/min) promoted a reduction of 49% in 120 min (p < 0.05). D Chiro-inositol at a bolus dose of 5 mg/kg had no effect. A single dose of 15 mg/kg produced a reduction of 21% (p < 0.05) in 120 min. D-Chiro-inositol (15 mg/kg) associated with manganese chloride (8.3 microg/min) decreased elevated plasma glucose 47% (p < 0.05) in 120 min. D-Chiro-inositol coadministered with manganese reduced glucose concentrations during the final 60 min (p < 0.05). 3-O Methyl-D-Chiro-inositol and D-chiro-inositol are components of the mediator structure. Manganese is also a presumed component of the mediator, having an important role in glucose uptake, insulin release and mediator generation. These compounds have also been identified in the literature as hypoglycemic agents. PMID- 10824708 TI - Newly diagnosed latent autoimmune diabetes in adults (LADA) is associated with low level glutamate decarboxylase (GAD65) and IA-2 autoantibodies. Diabetes Incidence Study in Sweden (DISS). AB - A quantitative assay with microSepharose was used to determine GAD65Ab and IA-2Ab levels in 771 population-based patients diagnosed with diabetes mellitus at 15 to 34 years of age, and in 828 matched controls. Among the patients, 587 (76%) were classified with type I, 108 (14%) with type II, and 76 (10%) with unclassifiable diabetes. The levels above normal demonstrated a prevalence of GAD65Ab in 66% of type I diabetes, 50% of type II diabetes and 54% of unclassifiable patients and for IA-2Ab in 40%, 17% and 21%, respectively. Among the autoantibody-positive sera, the LADA patients had a lower GAD65Ab index (median 0.19, p < 0.0001) and IA-2Ab index (median 0.28, p < 0.0001) than the type I patients (median 0.37 and 0.66). Patients with unclassifiable diabetes had a GAD65Ab (median 0.43) or IA 2Ab (median 0.63) index which was not different from the type I diabetes patients. Our data demonstrate that young adult new-onset LADA patients have low level GAD65Ab and IA-2Ab. The low-level autoantibodies may signify a less aggressive beta-cell autoimmunity, which may explain why these patients are often classified with type II or non-insulin-dependent diabetes. PMID- 10824709 TI - An anti-insulin serum, but not a glucagon antagonist, alters glycemia in fed chickens. AB - Attempts at altering plasma glucose and, as a consequence, food intake were performed in fed broiler chickens by single i.v. injection of des-His1(Glu9) glucagon amide (a glucagon antagonist) or a non-stimulating anti-insulin serum. Plasma glucose level was not altered by des-His1(Glu9) glucagon amide but was rapidly and largely increased (for at least 2 h) by the injection of the insulin immune serum. Hour and cumulative food intake were unaltered up to 10 h post injection. These results strongly suggest that in fed chickens, plasma glucose is mainly, if not exclusively, controlled by plasma insulin, and that the transient and heavy hyperglycemia evoked by inhibiting insulin action does not alter food intake. PMID- 10824710 TI - Chronic corticosterone treatment impairs Leydig cell 11beta-hydroxysteroid dehydrogenase activity and LH-stimulated testosterone production. AB - The effects of excess corticosterone on luteinizing hormone (LH)-stimulated Leydig cell testosterone production and activity of 11beta-HSD was studied. Adult male rats (200-250 g body weight) were treated with corticosterone-21-acetate (2 mg/100 g body weight, i.m., twice daily) for 15 days. Another set of rats was treated with corticosterone (dose as above) plus LH (ovine LH 100 microg/kg body weight, s.c., daily) for 15 days. Corticosterone administration significantly increased serum and testicular interstitial fluid (TIF) corticosterone but decreased testosterone levels. Administration of LH with corticosterone partially prevented the decrease in serum and TIF testosterone. The oxidative activity of 11beta-hydroxysteroid dehydrogenase (11beta-HSD) was significantly decreased in Leydig cells of rats treated with corticosterone alone and in combination with LH. The direct effect of corticosterone on Leydig cell steroidogenic potency was also studied in vitro. Addition of corticosterone to Leydig cell culture showed a dose dependent effect on LH-stimulated testosterone production. Corticosterone at 50 and 100 ng/ml did not alter LH-stimulated testosterone production, but at high doses (200-400 ng/ml), decreased basal and LH-stimulated testosterone production. Basal and LH-stimulated cAMP production was not altered by corticosterone in vitro. It is concluded from the present study that elevated levels of corticosterone decreased the oxidative activity of 11beta-HSD and thus resulting in impaired Leydig cell steroidogenesis and the inhibitory effects of corticosterone on testosterone production appear to be mediated through inhibition of LH signal transduction at post-cAMP level. PMID- 10824711 TI - Possible involvement of corticosterone in bone loss of genetically diabetic db/db mice. AB - The etiology of bone loss in non-insulin dependent diabetes mellitus is still unknown. We compared serum biochemical parameters and bone parameters of genetically diabetic db/db mice with those of their control non-diabetic +/+ mice. We found that serum corticosterone levels of the db/db mice were significantly elevated after 5 weeks while bone mineral density of femur metaphysis significantly decreased in the db/db mice after 12 weeks of age compared with age matched +/+ mice. To explore the causal relationship between the serum corticosterone levels and the bone loss, metyrapone (100 mg/kg, p.o., twice a day), a glucocorticoid synthesis inhibitor, was administered to these mice for 4 weeks after the age of 8 weeks. The compound significantly decreased serum corticosterone levels in both strains. Metyrapone prevented bone loss by increasing the bone mineral content of the metaphysis in the db/db mice. In addition, the treatment slightly improved the ratio of ash weight to dry weight in the db/db mice. These results suggest that increased serum corticosterone levels are concerned with the etiology of bone loss in non-insulin dependent diabetic db/db mice. PMID- 10824712 TI - Role of food intake in the modulation of hexarelin-induced growth hormone release in normal human subjects. AB - Hexarelin (HEX) is a new synthetic analog of the Growth Hormone releasing peptides and is stronger than GHRH in releasing GH in vivo. No information is available on the effect of food ingestion on HEX-induced GH secretion. On the other hand, we have previously demonstrated that food intake at lunchtime in normal subjects has an inhibitory effect on the GH response to GHRH. The aim of the present study was to investigate the effect of food ingestion on GH secretion induced by HEX as compared to GHRH in six normal men (aged 23-29 years) and six normal women (aged 24-29 years). The body weights for all subjects were within 120% of their ideal body weight, according to their sex and age. Our data confirm that HEX is much more powerful than GHRH in inducing GH release in humans, both in the fasting state (GH-AUC: 3010 +/- 695 after HEX, vs. 1339 +/- 281 after GHRH, microg/L/120 min; p<0.06) and after a meal (GH-AUC: 1523 +/- 121, after HEX, vs. 309 +/- 61, after GHRH, microg/L/120 min; p<0.06). Moreover, our study shows that food intake partially blunts the fasting GH response to HEX (GH-AUC: 3010 +/- 695 after HEX, in fasting state, vs. 1523 +/- 121 after HEX, after meal, microg/L/120 min; p<0.06; mean inhibition of AUC 41.02 +/- 7.96%), whereas it nearly abolishes the GH response to GHRH in the same subjects (GH-AUC: 1339 +/- 281 after GHRH, in fasting state, vs. 309 +/- 61 after GHRH, after meal, microg/L/120 min; p<0.06; mean inhibition of AUC 70.31 +/- 6.22%). In conclusion, our study confirms that HEX acts differently from GHRH; the GH releasing effect of HEX could be only partially influenced by the physiological metabolic or neuroendocrine food-related modifications. PMID- 10824713 TI - Salivary testosterone at high altitude in man. PMID- 10824714 TI - Short stature is common feature of mitochondrial gene abnormalities with diabetes. PMID- 10824715 TI - The security circuit: a proposed construct for the central nervous system. AB - The theory of the Security Circuit offers a new conceptual framework for the numerous brain functions pertaining to survival. Many such activities are related to the coordination of the conventional physiological systems in the maintenance of homeostasis and the defense of the internal environment. This work, however, focuses on that aspect of brain function that makes possible man's interaction with external environment, thereby facilitating individual and group adaptation and survival. In our schema we greatly enlarge the conceptualization of the autonomic nervous system. We postulate the existence of a central mechanism pertaining to the security and survival function. In its action the complex physiological ANS substrate designated the Security Circuit, is likened to a whirlpool bath, in which balance is maintained with respect to electrically charged particles rather than water. The use of the Jacuzzi (or whirlpool bath) format makes it possible to tie together the components of the triune brain (MacLean), which consists of R-complex, limbic system, and neocortex. The forebrain, in part, is viewed as the depository of biological symbolism for a huge number of elements, which vary with the individual. Among these are security and support figures, and others that make up meaningful relationships. These are seen as represented on posts, consisting of either individual, or else groupings, of neurons which are linked to the limbic system to trigger oft predictable patterns of behavior and/or emotional expression. The limbic system serves as the energizer in arousal-defense. It also serves to trigger instinctive and other psychomotor patterns in the pursuit of goals which have survival value, while simultaneously producing the chemistry behind emotions useful to man. Some of these psychomotor patterns (i.e., behavior) facilitate the provision of nutrients and warmth for the individual, and so ensure internal homeostasis, while others effectuate group action towards this end. The apparent neuroendocrinal sequence of events in many common environmental situations is discussed. Responses to a perceived threat are examined, noting both the reflex behavior patterns resulting from somatic and ANS discharge, and the associated emotional expression. The chemoelectrical workings of the limbic system keep the central mechanism in a state of balance, while neocortical centers of cognition translate the minute changes in the chemical mix into subjective emotional experience which contribute to the phenomenon of consciousness. The R-complex is seen as the physiological component of the unconscious brain regulating the internal vital functions. The task of the higher centers is not merely to inhibit impulsive (primitive) responses, but also has the following functions: (i) determine the more sophisticated, effective manner of dealing with perceived threats, and (ii) initiate and facilitate the pursuit of goals promoting survival and well-being for self and group. The human thrust to self-expression or self-actualization is seen as an outgrowth of this process. The theory suggests that intellectual and creative activity within a group is an extension in evolution of primal, survival functions. This thrust has resulted in the growth of culture and development of civilization. While the related neurophysiological activities enhance security and environmental adaptation for self and group, they simultaneously resolve heightened microelectrical tension within the underlying mechanism within the individual's neuroendocrinal complex. The brain, and autonomic and endocrine systems are seen as parts of the same physiological system which pertains to security, survival, and well-being. The concept is presented that the thrust to maintain microelectrical balance within the Security Circuit (a substrate of the CNS) has been the ever-present, ongoing driving force behind the evolution of the "higher," neocortical centers of consciousness during the psych PMID- 10824716 TI - Personality variables and compliance with insulin therapy in Type 2 diabetic subjects. AB - Forty-five diabetic patients were studied to evaluate adaptation and coping strategies. The authors have also analysed personality traits mainly to study different behaviour in compliance conduct. The results revealed an important psychological dimension made up of difficulties in accepting insulin immediately, in fear of addiction and doubts about the therapy. On the basis of these results the sample was then divided into two subgroups, which were then tested and compared with the Adjective Check List. The subgroup that showed more fear, insecurity and initial resistance towards insulin therapy appeared to be more rigid and seemingly conforming. These people also revealed personality aspects compatible with the presence of passive-aggressive and avoidant traits. PMID- 10824717 TI - Influence of acarbose on post-prandial insulin requirements in patients with Type 1 diabetes. AB - The primary objective of this double-blind, placebo-controlled, randomised cross over study was to investigate the influence of acarbose on insulin requirement in patients with Type 1 diabetes (T1DM) following a standardised meal. In addition, the study assessed the effects of acarbose on post-prandial triglyceride, glucagon and gastrointestinal peptide levels, gastric emptying, and oxidative glucose metabolism. Following normalisation of their blood glucose, 10 patients received a standardised meal together with acarbose (100 mg) or placebo. Each patient was evaluated twice (separated by 10+/-3 days), and the cross-over study design ensured that they received both acarbose and placebo. The insulin requirement for maintenance of normoglycaemia was assessed using a closed-loop insulin infusion system (artificial pancreas, Biostator). Acarbose produced a statistically significant reduction in mean insulin requirement over a 3-hr period following the meal compared with placebo (5171.7+/-2282.6 mU vs 8074.5+/ 3045.4 mU; p=0.003). The level of blood glucose control over the same period was similar in the two groups. Gastric inhibitory polypeptide levels also showed a statistically significant decrease with acarbose treatment compared with placebo for AUC (area under the curve; p=0.006) and Cmax (maximum plasma concentration; p=0.022), but not tmax (time to reach Cmax from the start of the standardised meal; p>0.05). Analysis of the other efficacy parameters revealed no statistically significant differences between acarbose treatment and placebo (p>0.05). These results indicate that acarbose decreases insulin requirement in patients with T1DM without affecting gastric emptying. PMID- 10824718 TI - Glycaemic and insulinaemic indices of Mexican foods high in complex carbohydrates. AB - For the purpose of enriching the knowledge on the glycaemic (GI) and insulinaemic (InIn) indices of indigenous foods, 3 single foods and 3 realistic high complex carbohydrate meals (bread=100) were studied in 8 healthy subjects. Observed GI (mean+/-SEM) were: beans (B) 19.3+/-3.4, wheat tortilla (WT) 42.5+/-6.9, corn tortilla (CT) 73.8+/-6.5, wheat tortilla beans taco (BWT) 39.9+/-12.8, corn tortilla beans taco (BCT) 56.2+/-8.2 and corn tortilla potato taco (PT) 111.0+/ 11.5. The glycaemic index of all foods was lower than white bread (WB) (p<0.01) except for PT. B represented the lowest GI. InIn indices of B (41.7+/-4.5), WT (62.4+/-6.7) and CT (85.5+/-7.8) were lower than WB (p<0.01), whereas PT (171.3+/ 11.4) was higher. B InIn was the lowest except for WT, and the highest InIn was for PT. B, WT and CBT might be included in a regular diet for weight maintenance and control of hunger. It is suggested to moderate the ingestion of CT and PT in subjects with obesity and hyperglycaemia complications. It is important to study single and realistic foods with varied composition in diabetic subjects, whose glycaemic responses in reference to their insulinaemic responses might be different to those reported in the present study. PMID- 10824719 TI - Protein wasting in severe illness: pathogenesis and therapy. PMID- 10824720 TI - Role of glutamine depletion in severe illness. PMID- 10824721 TI - Protein metabolism during space flight. PMID- 10824722 TI - Trends in tracer techniques, including use of positron emission tomography. PMID- 10824723 TI - Methods for measuring polymerisation biosynthesis: three general solutions to the problem of the "true precursor". PMID- 10824724 TI - From syndromes to specific disease mechanisms. The search for the causes of myocardial infarction. PMID- 10824725 TI - Assessment of coronary vasomotor function: old and new tools. AB - Atherosclerosis has an impact on the vasomotor reaction of coronary segments to iodinated non-ionic contrast agents. Angiographically normal coronary segments show divergent vasomotor reactions to iodixanol or iopromide according to the presence of, and distance from, a coronary atherosclerotic lesion. The mechanism responsible for the above-mentioned vasomotor effect does not seem to involve flow-mediated vasodilation or endothelial nitric oxide synthesis. On the other hand, a cyclooxygenase product may be, at least in part, responsible for the vasodilating effect of non-ionic agents on epicardial coronary arteries. These findings have potential clinical implications that are herein discussed. PMID- 10824726 TI - The endothelium-derived hyperpolarizing factor: does it play a role in vivo and is it involved in the regulation of vascular tone only? AB - Several investigations performed in vitro have shown that vascular endothelia can release diffusible compounds capable of inducing hyperpolarization of the smooth muscle fibers. Experiments in vitro have shown that these compounds can cause coronary vasodilation and alter cardiac performance. Experiments in vivo only showed the occurrence of vasodilation. While it has been shown that the release of these endothelium-derived hyperpolarizing factors (EDHFs) is not impaired by the inhibition of nitric oxide synthase and cyclooxygenase, the precise nature of the compound(s) has not yet been identified. It is possible that they vary depending on the organ and animal species. However, a common feature of the activity of EDHFs is the activation of calcium-dependent potassium channels, inhibitable by charybdotoxin and apamin. Furthermore in the coronary circulation of many species EDHF seems to be a cytochrome P450-dependent non-prostanoid metabolite of arachidonic acid activated by a number of chemical and physical stimuli similar to those which are known to activate endothelial nitric oxide synthase. Using compounds which inhibit cytochrome P450 and blockers of the calcium-dependent potassium channels, researchers can study the physiological and pathophysiological relevance of EDHF in vivo thus disclosing the potential therapeutic applications of the basic knowledge in this field. PMID- 10824727 TI - Assessment of myocardial performance with ventricular pressure-volume relations: clinical applications in cardiac surgery. AB - The assessment of myocardial performance in patients with cardiomyopathy is of vital importance in cardiology and cardiac surgery, especially considering the significant increase in the number of patients treated for congestive heart failure. Left ventricular pressure-volume analysis is a method, which can assess accurately myocardial contractility, separating the systolic and diastolic function at different preload and afterload conditions. This technique can be used for determination of the efficacy of a therapeutic pharmaceutical or surgical intervention, for instance the assessment of ventricular function after coronary revascularization. A few studies using the conductance catheter for the analysis of ventricular pressure-volume relations in the field of cardiac surgery have been published. In our center we started to use this technique to analyze cardiac surgical procedures, like mitral valve reconstruction, aortic valve replacement, myocardial revascularization, left ventricular assist, and surgical left ventricular remodeling. This information will be used to develop a therapeutic strategy, which may optimize surgical indications and improve the peri- and postoperative treatment and the efficacy of that surgical technique. In this short review the possible clinical use in cardiac surgery and the methodology of the pressure-volume loops have been described. Three clinical cases are presented to demonstrate functional information related to the surgical treatment of congestive heart failure patients. PMID- 10824728 TI - Doppler-derived acceleration rate of right ventricular early filling reliably predicts mean right atrial pressure at baseline and after loading manipulations in patients with chronic heart failure. AB - BACKGROUND: We investigated whether Doppler-derived variables of tricuspid flow could estimate mean right atrial pressure and monitor its changes after loading manipulations in patients with chronic heart failure. METHODS: Simultaneous mean right atrial pressure (Swan-Ganz catheterization) and tricuspid Doppler recordings were initially evaluated in 136 patients (23 with atrial fibrillation) with chronic heart failure and severe left ventricular systolic dysfunction, and then were repeated in 18 patients after unloading (sodium nitroprusside infusion) and in 13 patients after overloading (active leg elevation) manipulations. RESULTS: A significant correlation was observed between mean right atrial pressure and peak E velocity (r = 0.70), early deceleration time (r = -0.72) and acceleration time (r = -0.75). However, the best correlation found was between the acceleration rate of early flow and mean right atrial pressure, and it was identical in patients in sinus rhythm or with atrial fibrillation (r = 0.98). Moreover, after acute effective unloading or overloading manipulations, although all Doppler tricuspid variables changed significantly, the acceleration rate of early flow still emerged as the strongest independent predictor of mean right atrial pressure (r = 0.95 and 0.99, respectively). CONCLUSIONS: Doppler-derived acceleration rate of early diastolic tricuspid flow is a powerful tool to predict mean right atrial pressure and to monitor its changes after loading manipulations. PMID- 10824729 TI - Clinical correlates of elevated plasma natriuretic peptides and Big endothelin-1 in a population of ambulatory patients with heart failure. A substudy of the Italian Network on Congestive Heart Failure (IN-CHF) registry. IN-CHF Investigators. AB - BACKGROUND: Activation of neuroendocrine factors plays a major role in the pathophysiology and progression of heart failure. The aim of the present study was 1) to assess the clinical correlates of elevated plasma natriuretic peptides [atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP)] and Big endothelin-1 in a population of 180 ambulatory patients from the Italian registry of heart failure (Italian Network on Congestive Heart Failure, IN-CHF) in 22 clinical centers, 2) to assess the within-patient variability of plasma BNP concentration, and 3) to evaluate the analytical agreement for BNP determination between a core laboratory and local sites. METHODS: ANP and BNP were measured with specific immunoradiometric methods, Big endothelin-1 with an enzyme immunoassay. RESULTS: Elevated BNP was associated with severe mitral valve regurgitation (odds ratio 8.546, 95% confidence interval 1.879-38.510, p = 0.0052); high circulating concentrations of ANP and BNP were found in older patients, and in patients with higher NYHA functional class or reduced left ventricular ejection fraction. Elevated plasma concentration of Big endothelin-1 was a strong and independent predictor of atrial fibrillation (odds ratio 4.001, 95% confidence interval 1.531-10.454, p = 0.0047). Plasma concentration of BNP was reasonably stable at 3-month interval in patients with mild-to-moderate heart failure (mean between-visit difference -1.5+/-45 pg/ml, n = 96). There was a satisfactory analytical agreement between the central laboratory and sites, over a broad range of concentrations (2-1133 pg/ml, n = 283) with a slope for the best line fitted by linear regression of 1.09 (r2 = 0.96). CONCLUSIONS: BNP assay may become an appropriate tool for routine clinical practice in patients with congestive heart failure. PMID- 10824730 TI - Reduction of coronary flow reserve non-invasively determined by transthoracic Doppler echocardiography as a predictor of left anterior descending coronary artery stenosis. AB - BACKGROUND: The aim of this study was to evaluate if a reduced coronary flow reserve determined by transthoracic echocardiography alone or combined with contrast agents may represent a predictive index of significant left anterior descending coronary artery (LAD) stenosis. METHODS: Thirty-four patients (mean age 59+/-9 years) undergoing coronary angiography for coronary artery disease were studied. Coronary stenosis was classified (according to visually determined percent narrowing) as severe (> 75%), moderate (40 to 75%) and mild (< 40%). Coronary blood flow velocities were recorded in each patient at baseline and after low-dose dipyridamole administration by use of a 3.5 MHz transducer with a machine equipped with second harmonic capability and nondirectional color Doppler software. Coronary flow reserve was defined as the ratio of hyperemic to basal diastolic peak velocity. RESULTS: Adequate Doppler recordings in the LAD were obtained by transthoracic echocardiography in 26/34 patients (76%); the infusion of Levovist allowed for the visualization of LAD flow in a further 7 patients, with an overall feasibility of 97%. Coronary flow reserve was significantly higher in the group of patients with mild coronary lesions (2.3+/-0.3) than in patients with moderate (1.68+/-0.29, p = 0.0004) or severe (1.49+/-0.39, p = 0.0005) LAD stenosis. CONCLUSIONS: By use of transthoracic echocardiography combined with contrast agents it is possible to visualize blood flow velocities in the LAD and to evaluate coronary flow reserve after dipyridamole infusion with a non-invasive approach. Combined with angiographic findings, this diagnostic approach could be useful in giving additional information to assess the functional significance of a stenotic coronary lesion. PMID- 10824731 TI - Relation of left ventricular chamber and midwall function to age in normal children, adolescents and adults. AB - BACKGROUND: The goal of this study was to identify the effect of body growth and aging, and normal limits of the relation of left ventricular endocardial and midwall shortening to wall stress. METHODS: Endocardial and midwall shortening and circumferential end-systolic stress were assessed in 388 normotensive, normal weight adults (226 men, 162 women, age 18 to 85 years) and 332 children and adolescents (180 males, 152 females, age 4 to 17 years) by two-dimensional targeted M-mode echocardiography and cuff blood pressure measurements. RESULTS: End-systolic stress decreased with age in children and adolescents (p < 0.001), but not during adulthood and maturity. The negative relation of endocardial shortening to end-systolic stress was stronger in adults than in children and adolescents (slope difference p < 0.005). The negative relation of midwall shortening to end-systolic stress was negligible in children and adolescents (r = -0.07, p = 0.18), whereas it was more evident, although weak, in adults (r = 0.14, p < 0.007). For a given level of end-systolic stress, endocardial shortening decreased by 0.32%/year in children and adolescents (multiple r = 0.51, p < 0.0001) and by 0.05%/year in adults, whereas midwall shortening decreased by 0.26%/year during body growth and by 0.02%/year in adults. CONCLUSIONS: In the presence of normal blood pressure and normal weight, the relations between left ventricular wall stress and both chamber and myocardial function are weakly but significantly influenced by age. Left ventricular chamber function is markedly influenced by wall stress, while this influence is reduced for left ventricular wall mechanics. PMID- 10824732 TI - Isolated noncompaction of the myocardium: an exceedingly rare cardiomyopathy. A case report. AB - Isolated noncompaction of the left ventricular myocardium is a rare cardiac disorder due to an arrest in myocardial morphogenesis. It is characterized by prominent and excessive trabeculation in a ventricular wall segment, with deep intertrabecular spaces perfused from the ventricular cavity. Echocardiographic findings are important clues for the diagnosis. Clinical symptoms include signs of left ventricular systolic dysfunction even to the point of heart failure, ventricular arrhythmias, and embolic events. We describe an adult case in whom the only clinical symptoms were life-threatening ventricular arrhythmias. Transthoracic echocardiography did not contribute to the diagnosis, which was made thanks to left ventricular contrast angiography. Electrophysiological testing induced a fast monomorphic sustained ventricular tachycardia, with hemodynamic impairment, that was refractory to pharmacological treatment, and for this reason a permanent cardioverter-defibrillator was implanted. A subsequently performed transesophageal echocardiographic examination showed a localized, regional increase in left ventricular wall thickness and degree of trabeculation. The causes and electrophysiological mechanisms of arrhythmias in noncompaction are still unknown: grossly irregular branching and connecting of myocardial fascicles in the noncompacted segments, isometric contraction with increased wall stress, and localized coronary perfusion impairment can all induce disorganized or delayed activation and increase the potential for arrhythmias. This is the first reported case of noncompaction in which an implantable defibrillator was used to control life-threatening arrhythmias. PMID- 10824733 TI - Woven coronary artery: differential diagnosis with diffuse intracoronary thrombosis. AB - Woven coronary malformation is characterized by the branching of a major epicardial coronary artery into thin channels which then merge again in a normal conduit. The angiogram can suggest a filling defect instead of a malformation and an undue coronary angioplasty could be performed determining some damage to the arterial wall. In this case report we describe a patient with a stenosis on the left anterior descending coronary artery and a woven coronary artery on the right coronary artery. In 1995 a coronary angioplasty was performed on the left anterior descending coronary artery. Four years later a coronary angiogram did not show any changes in the right coronary artery. In this patient the malformation did not induce any reduction in the coronary reserve as shown at cardiac scintigraphy. We need more information about the natural history of such a malformation. PMID- 10824734 TI - Recent developments in the cell biology and biochemistry of glycosylphosphatidylinositol lipids (review). AB - Glycosylphosphatidylinositols (GPIs) represent an abundant and ubiquitous class of eukaryotic glycolipids. Although these structures were originally discovered in the form of GPI-anchored cell surface glycoproteins, it is becoming increasingly clear that a significant proportion of the GPI synthetic output of a cell is not directed to protein anchoring. Indeed, pools of non-protein-linked GPIs can approach 10(7) molecules per cell in some cell types, especially the protozoa, with a large proportion of these molecules being displayed at the cell surface. Recent studies which form the subject of this review indicate that there is (a) considerable diversity in the range of structural modifications found on GPI glycolipids within and between species and cell types, (b) complexity in the topological arrangement of the GPI biosynthetic pathway in the endoplasmic reticulum, and (c) spatial restriction of the biosynthetic pathway within the endoplasmic reticulum. Furthermore, consistent with additional functional roles for these lipids beyond serving as protein anchor precursors, products of the GPI biosynthetic pathway appear to be widely distributed in the cellular endomembrane system. These studies indicate that there is still much to learn about the organization of glycolipid biosynthetic pathways in eukaryotic cells, the nature and subcellular distribution of the lipid products of these pathways, and the function of these lipids within cells. PMID- 10824735 TI - Single molecule microscopy of biomembranes (review). AB - Recent advances in the development of new microscopy techniques with a sensitivity of a single molecule have gained access to essentially new types of information obtainable from imaging biomolecular samples. These methodologies are analysed here in terms of their applicability to the in vivo visualization of cellular processes on the molecular scale, in particular of processes in cell membranes. First examples of single molecule microscopy on cell membranes revealed new basic insight into the lateral organization of the plasma membrane, providing the captivating perspective of an ultrasensitive methodology as a general tool to study local processes and heterogeneities in living cells. PMID- 10824736 TI - Tilted peptides: a motif for membrane destabilization (hypothesis). AB - Cell life depends on the dynamics of molecular processes: molecule folding, organelle building and transformations involving membrane fusion, protein activation and degradation. To carry out these processes, the hydrophilic/hydrophobic interfaces of amphipathic systems such as membranes and native proteins must be disrupted. In the past decade, protein fragments acting in the disruption of interfaces have been evidenced: they are named the tilted or oblique peptides. Due to a peculiar distribution of hydrophobicity, they can disrupt hydrophobicity interfaces. Tilted peptides should be present in many proteins involved in various stages of cell life. This hypothesis overviews their discovery, describes how they are detected and discusses how they could be involved in dynamic biological processes. PMID- 10824737 TI - Insulin stimulates the release of a subset of GPI-anchored proteins in a G protein independent manner. AB - The glycosyl-phosphatidylinositol anchored protein, membrane dipeptidase (EC 3.4.13.19) is released from the surface of 3T3-L1 adipocytes in response to insulin treatment through the action of a phospholipase C. The present study investigates the role of guanine-nucleotide binding proteins (G-proteins) in this process. Treatment of permeabilized 3T3-L1 adipocytes with GTPgammaS did not cause release of membrane dipeptidase into the medium, while GDPbetaS did not inhibit the insulin-stimulated release of membrane dipeptidase. Other activators of G-proteins, including the tetradecapeptide mastoparan, pertussis toxin and AlF3, also caused no significant release of membrane dipeptidase from the surface of the 3T3-L1 adipocytes. From these observations it is concluded that G-proteins are not involved in the insulin-stimulated release of membrane dipeptidase. Although X-Pro aminopeptidase (EC 3.4.11.9) is GPI-anchored in 3T3-L1 adipocytes as shown by digestion with bacterial phosphatidylinositol-specific phospholipase C, it was not released upon insulin treatment of the cells, indicating that only a subset of the GPI-anchored proteins are susceptible to insulin-stimulated release. PMID- 10824738 TI - Amino acid residues N450 and Q449 are critical for the uptake capacity and specificity of UapA, a prototype of a nucleobase-ascorbate transporter family. AB - Specific carrier-mediated transport of purine and pyrimidine nucleobases across cell membranes is a basic biological process in both prokaryotes and eukaryotes. Recent in silico analysis has shown that the Aspergillus nidulans (UapA, UapC) and bacterial (PbuX, UraA, PyrP) nucleobase transporters, and a group of mammalian L-ascorbic acid transporters (SVCT1 and SVCT2), constitute a unique protein family which includes putative homologues from archea, bacteria, plants and metazoans. The construction and functional analysis of chimeric purine transporters (UapA-UapC) and UapA-specific missense mutations in A. nidulans has previously shown that the region including amino acid residues 378-446 in UapA is critical for purine recognition and transport. Here, we extend our studies on UapA structure-function relationships by studying missense mutations constructed within a 'signature' sequence motif [(F/Y/S)X(Q/E/P)NXGXXXXT(K/R/G)] which is conserved in the putative functional region of all members of the nucleobase/ascorbate transporter family. Residues Q449 and N450 were found to be critical for purine recognition and transport. The results suggest that these residues might directly or indirectly be involved in specific interactions with the purine ring. In particular, interaction of residue 449 with C-2 groups of purines might act as a critical molecular filter involved in the selection of transported substrates. The present and previous mutagenic analyses in UapA suggest that specific polar or charged amino acid residues on either side of an amphipathic alpha-helical transmembrane segment are critical for purine binding and transport. PMID- 10824739 TI - Membranes, sensors and cell surfaces. PMID- 10824740 TI - Primary bile reflux gastritis: which treatment is better, Roux-en-Y or biliary diversion? AB - Primary bile reflux gastritis is an unusual and elusive problem. Postgastrectomy bile reflux has been long recognized and treated variously with Roux-en-Y gastrojejunostomy, Braun enteroenterostomy, and Henley jejunal interposition. All of these procedures have been fraught with postoperative side effects, the worst of which is stasis. A new procedure utilizing biliary diversion has been proposed to divert bile from the gastric lumen without vagotomy or gastric resection. This procedure was used for 16 patients with diagnosed bile reflux, and results were compared with those of a previous group of 21 patients who had been treated with Roux-en-Y gastrojejunostomy. The patient groups were similar in age, sex, weight, symptoms, and results of investigative studies. The earlier group all had vagotomy, antrectomy, and gastrojejunal anastomosis to a 45-cm Roux limb. The later group all had an end-to-side choledochojejunostomy to a 45-cm Roux limb, taken 45 cm from the ligament of Treitz. The patients in the bile diversion group had fewer complications and shorter hospital stays. In addition, they had few postoperative complaints, no further operations for either bile reflux or upper gastrointestinal stasis, and no long-term deaths due to gastrointestinal problems or malnutrition. Their eventual postoperative gastric emptying improved significantly when compared with the Roux-en-Y patients, suggesting that the dysmotility observed preoperatively may well have been a result of the bile injury to the stomach, rather than an underlying gastric dysmotility. PMID- 10824741 TI - Management of choledocholithiasis in the era of laparoscopic surgery. AB - Laparoscopic biliary surgery is changing the management of choledocholithiasis. Between November 1989 and December 1998, 2834 cholecystectomies were performed at two institutions. Choledocholithiasis was suspected in 420 patients on the basis of elevated preoperative laboratory and ultrasound criteria [bilirubin, alkaline phosphatase, serum glutamic-oxaloacetic transaminase, serum glutamate pyruvate transaminase, and common bile duct (CBD) size]. One hundred seventeen patients had preoperative endoscopic retrograde cholangiopancreatography (ERCP) because of persistent elevation in their enzymes beyond 24 to 48 hours or as an emergency. Laparoscopic intraoperative cholangiogram was attempted in 329 patients whose enzymes fell rapidly within the first 24 to 48 hours or had a failed ERCP. Eighty one of the 329 were found to have stones. Seventy-three had laparoscopic attempt to clear the CBD, with success in 62 patients (85%). This included 41 transcystic duct and 21 direct CBD exploration. Eight patients had post-operative ERCP for retained stones. Six (0.25%) were in patients with normal preoperative enzymes. We conclude that choledocholithiasis can be suspected with preoperative laboratory and ultrasound criteria. By waiting 24 to 48 hours (except in an emergency), a good number of CBD stones will pass. With increases in laparoscopic experience, laparoscopic removal of CBD stones may replace preoperative ERCP. The small number of cases of retained or missed stones that occur with the use of selective cholangiography can be easily handled with postoperative ERCP. PMID- 10824742 TI - The role of positron emission tomography in selecting patients with metastatic cancer for adrenalectomy. AB - Metastases to the adrenal glands usually signal disseminated disease. However, isolated metastases do occur that may be curable with adrenalectomy. Functional imaging with positron emission tomography (PET) can differentiate benign from malignant pathology and isolated from disseminated metastases. The purpose of this study was to determine whether PET scanning can influence the outcome of adrenalectomy for metastatic disease. We conducted a retrospective review of eight patients undergoing adrenalectomy for presumed isolated metastatic disease from 1985 through 1997. The patients included six women and two men with an average age of 58 (range, 36-74). Their primary tumors were six lung carcinomas, one renal cell carcinoma, and one colon carcinoma. The adrenal masses were located on the right in six patients, on the left in one, and bilaterally in one. Before operation, all patients were evaluated by chest and abdominal CT. Four patients were also evaluated by PET scan. Six right, one left, and one bilateral adrenalectomies were performed. Associated organ resections included two right partial nephrectomies and one right total nephrectomy, one left partial nephrectomy, two distal pancreatectomies, one splenectomy, and two partial hepatic resections. All eight patients survived operation. There were no major perioperative complications, but one patient required readmission for congestive heart failure. Three of the four patients who did not have PET scanning died from 4 to 48 months after operation with disseminated disease from lung, colon, and renal carcinoma respectively. The remaining patient who did not have PET scanning is alive and well 11 years later. Two of the four patients who had PET scans showing isolated disease are alive at 28 and 43 months after operation, whereas the other two died of disseminated disease at 29 and 36 months after operation. We conclude that 1) adrenalectomy can provide survival benefit in patients with isolated metastases, and 2) PET scanning is useful in confirming isolated metastatic disease and selecting patients for adrenalectomy. PMID- 10824743 TI - Changing patterns of care for occult breast lesions in a community teaching hospital. AB - We performed a retrospective analysis of 384 consecutive stereotactic breast biopsies (SBBs) from March 1995 through January 1999 and compared it with our historical breast biopsy experience. Two hundred forty-four patients underwent biopsies for microcalcifications and 135 patients for abnormal mammographic densities. Pathology diagnoses included 302 patients with benign disease, 35 patients with atypical ductal hyperplasia, 4 patients with lobular carcinoma in situ, 29 patients with ductal carcinoma in situ, and 9 patients with invasive breast cancer. These diagnostic rates were compared with our prior needle localized pathology findings. For the study period, the number of mammograms, open biopsies, and needle-localized biopsies remained stable. The number of SBBs, however, increased progressively in every year. Medicare reimbursement for SBB was $921.19, and for breast biopsy after needle localization, $1566.22. Our study strongly suggests that the availability of SBB has significantly lowered the threshold for recommending biopsy of abnormal mammograms. The increased utilization of SBB almost certainly indicates an increase in the overall cost of breast care. This cost must be balanced against substantial potential benefits of this minimally invasive technique: possible earlier diagnosis of atypical and precancerous lesions, patient reassurance in cases of uncertain mammographic interpretation, and a reduced need for follow-up of indeterminate mammograms. PMID- 10824744 TI - Seroma formation after breast cancer surgery: incidence and predicting factors. AB - The pathophysiology of seroma formation has yet to be determined. Therefore, the present study was undertaken to calculate the incidence of postoperative seromas after definitive breast cancer operations utilizing electrocautery dissection. Additionally, we attempted to identify risk factors associated with seroma development and to examine seroma formation in relation to operative procedure. A retrospective review of 252 breast cancer operations was undertaken. Patients were subdivided by operative procedure: modified radical mastectomy (MRM; n = 148), breast preservation with axillary node dissection (n = 64), or MRM with immediate reconstruction (n = 40). Electrocautery was used in development of skin flaps. Seromas developed in 39 of the 252 operations for an incidence of 15.5 per cent. Seroma formation was significantly lower in those patients receiving MRM with immediate reconstruction than in those receiving MRM (2.5% vs 19.6%; P = 0.009) and tended to be lower than for patients receiving breast preservation with axillary node dissection (14.06%; P = 0.052). Neoadjuvant chemotherapy was performed in 18 patients, of whom 6 developed seromas (P = 0.030). The incidence of postoperative seromas was low despite the use of electrocautery. An association of postoperative seromas with neoadjuvant chemotherapy was noted. Additionally, it appears that immediate reconstruction may reduce the incidence of postoperative seromas, presumably by filling the dead space in the chest wall. PMID- 10824745 TI - Adjuvant radiation after modified radical mastectomy for breast cancer fails to prolong survival. AB - Recent literature has reported improved local disease control and overall survival in premenopausal node-positive (stage II, and III) breast cancer patients undergoing modified radical mastectomy (MRM) using radiation therapy (RT) combined with chemotherapy. To assess the efficacy of postoperative RT in our own community, we analyzed all patients undergoing MRM for carcinoma utilizing an extensive database from the three major teaching hospitals in Tulsa, OK, between 1965 and 1993. A total of 5257 patients underwent MRM during this time period. One hundred thirty-seven patients were excluded for insufficient data or because they were found to be at stage IV, leaving a total study population of 5125. Overall survival (OS), overall mean survival (MS), disease free survival (DFS), and locoregional DFS (LRDFS) were analyzed for all patients and were further analyzed according to stage, lymph node involvement, and menopausal status. Median follow-up was 103 months. Statistical analysis was performed using Kaplan-Meier and t-tests. The DFS at 10 years was 65 per cent in the RT group and 80 per cent in the patients who did not receive RT (P = 0.00). No improved DFS was obtained in the radiation-treated patients, regardless of stage, lymph node involvement, or menopausal status. Similarly, the LRDFS at 10 years was 91 per cent in the RT group and 96 per cent in the patients who did not receive RT (P = 0.00). No improved LRDFS was obtained in the radiation-treated patients, regardless of stage, lymph node involvement, or menopausal status. The overall MS was 97 months in the RT group and 104 months in the patients who did not receive RT (P = 0.00). Comparisons of overall MS rates revealed apparent survival benefits from RT in the premenopausal node-negative group, postmenopausal one to four-positive-node group, and all stage I patients. This apparent survival advantage was not confirmed by Kaplan-Meier curves of OS. No other overall MS differences were detected according to stage, lymph node, or menopausal status. Using Kaplan-Meier survival curves, the OS in the RT group at 10 years was 46 per cent, and 63 per cent in the patients who did not receive RT (P = 0.00). No improved OS was obtained in the radiation-treated patients, regardless of stage, lymph node involvement, or menopausal status. These findings from a large breast cancer database failed to demonstrate any meaningful benefit from RT after MRM and serve to further question the efficacy of this treatment modality in postmastectomy breast cancer patients. PMID- 10824746 TI - Suppression of the invasive capacity of human breast cancer cells by inhibition of urokinase plasminogen activator via amiloride and B428. AB - Inhibition of urokinase plasminogen activator (uPA) has been shown to suppress cancer cell invasion and metastasis in the laboratory setting by numerous investigators. Most studies have used murine cell lines implanted in syngeneic rodents or transfected human cell lines grown in immunocompromised laboratory hosts. In this study using Matrigel invasion chambers and two separate uPA inhibitors, amiloride and B428, the invasive capacity of unaltered human breast cancer cells was significantly suppressed. Cell proliferation was also suppressed to a lesser degree. These findings suggest that uPA inhibition may be a valid clinical approach to the control of the invasion and metastasis of human cancers. PMID- 10824747 TI - Five-year experience with the "four-before" laparoscopic ventral hernia repair. AB - Between 1993 and 1998, we performed a linear study of laparoscopic ventral hernia repair performed in a standard fashion using expanded polytetrafluoroethylene on 49 patients. Eighteen patients had recurrent hernias and 30 patients were morbidly obese with a body mass index >30. Conversion to open procedure was required in two patients. Patients were observed a mean of 27 months. Three patients died of unrelated causes during the observation period. Three patients developed recurrent hernias. By a follow-up survey, we found that 90 per cent of patients were "satisfied" with their operation and results. Because of decreased complications, postoperative pain, hospital stay, and hernia recurrence, the "four-before" laparoscopic repair is our preference for ventral hernias. It has been particularly useful for obese patients and patients with recurrent ventral hernias. PMID- 10824748 TI - How well do third-year medical students learn key objectives in a case-based surgical lecture series? AB - Faculty members were asked to list major and minor concepts of their case-based session presented during the 12-week 3rd-year surgical clerkship. After each session, students were queried to list the key concepts presented. Data were collected from two groups: one at the end of an academic year and a second at the beginning of the next academic year. Faculty members listed a median of 10 major and 15 minor concepts. The mean number of matched major concepts ranged from 0.2 to 4, and from 0.2 to 3.4 for minor concepts. In a comparative analysis, the end of-the-year students listed a higher number of matched concepts for 17 of the 20 sessions than the beginning of the year students (8 sessions reached statistical significance, P < 0.05). The current case-based teaching method is not effective in emphasizing key concepts to students. Reformatting cases to better align with key concepts may be one solution to enhance a student's ability to grasp key concepts. Students at the end of the academic year outperformed those at the beginning of the year. This additional variable needs to be considered by faculty and incorporated into their teaching techniques. PMID- 10824749 TI - Incidence and significance of lobar atelectasis in thoracic surgical patients. AB - Lobar atelectasis, defined by complete lobar collapse and mediastinal shift on chest roentgenogram, represents one extreme form of postoperative atelectasis. We have evaluated the incidence and clinical significance of lobar atelectasis in a thoracic surgical patient group. A retrospective review was done of patients who underwent pulmonary resection over a 2-year period to determine patient characteristics, contributing comorbidities, and associated perioperative care factors. Lung resections were performed for both benign and malignant disease through open or video-assisted techniques. One hundred eighty patients had pulmonary resection, 101 males and 79 females, and they were divided into three groups: I, no complications (112 patients, 62%); II, complications unrelated to lobar atelectasis (60 patients, 33%); and III, complications of lobar atelectasis (8 patients, 5%). There was one death in the series, in the lobar atelectasis group (III). Mean age for the entire group was 64.5 +/- 12.5 years; however, patients in Groups II (67.3 years) and III (69.6 years) were significantly older than in Group I (P < 0.02). Mean hospital length of stay in Group I was 6 +/- 3 days, whereas that in Group II was 13 +/- 12 days (P < 0.001), and in Group III it was 27 +/- 31 days (P < 0.001). In addition, patients who developed lobar atelectasis were more likely to be male (88% vs 48%, P = 0.034), had a longer ICU length of stay (P < 0.001), were more likely to have two or more comorbidities (P < 0.05), and had a lower forced expiratory volume in 1 second (2.34 +/- 0.90 vs 1.96 +/- 0.63). All patients in the lobar atelectasis group were operated on for malignancy, but this was not significantly different from the other groups. None of the 16 patients who had thoracoscopy developed lobar atelectasis, but this also was not a significant finding. We conclude that severe postoperative atelectasis occurs as lobar atelectasis in approximately 5 per cent of patients who undergo pulmonary resection and significantly adds to the intensive care unit and hospital length of stay. The etiology of lobar atelectasis appears to be multifactorial and warrants further study to define mechanisms of occurrence and their prevention. PMID- 10824750 TI - The incidence of splenectomy is decreasing: lessons learned from trauma experience. AB - Over the past decade, splenic preservation has become a well-reported and accepted principle in trauma management. The reasons for splenic preservation may have influenced nontraumatic surgical management as well. To investigate the changing incidence and indications for splenectomy, we conducted a 10-year review of all splenectomies at our institution. During this time, between January 1, 1986, and December 31, 1995, 896 patients underwent splenectomy. Hospital charts and records were examined to determine the etiology and incidence of splenectomy. Indications were classified as: 1) trauma, i.e., performed for blunt or penetrating injury; 2) hematologic malignancy, i.e., therapy or staging of underlying leukemia, Hodgkin's lymphoma, or non-Hodgkin's lymphoma; 3) cytopenia, i.e., treatment of thrombocytopenia, anemia, or leukopenia; 4) iatrogenic, i.e., injury during another procedure; 5) incidental, i.e., required for adjacent organ resection; 6) portal hypertension, i.e., left-sided portal hypertension or during shunting procedure; 7) diagnostic, i.e., uncertainty excluding hematologic malignancy; or 8) other, i.e., miscellaneous indications. Trauma accounted for 41.5 per cent of all splenectomies during this time period, hematologic malignancy 15.4 per cent, cytopenia 15.6 per cent, incidental 12.3 per cent, iatrogenic 8.1 per cent, portal hypertension 2.3 per cent, diagnostic 2.0 per cent, and other 2.7 per cent. Comparing the first and second 5-year time periods, the following increases/decreases in average annual incidence were noted: splenectomy for all indications, -36.9 per cent; trauma, -32.9 per cent; hematologic malignancy, -51.4 per cent; cytopenia, 35.1 per cent; incidental, 35.9 per cent; iatrogenic, -30.2 per cent; diagnostic, +4.9 per cent, and other, 57 per cent. Traumatic injury to the spleen remains the most common indication for splenectomy, but the incidence has decreased dramatically over the past 10 years. Splenectomies for treatment of hematologic malignancies and cytopenia, as well as incidental and iatrogenic splenectomies, have also decreased significantly. Only the incidence of diagnostic splenectomy has remained stable. Although initiated within the field of trauma, the advantages of splenic preservation now appear to be well recognized beyond that field. PMID- 10824751 TI - Cervico-mediastinal extension of thyroid cancer. AB - A surgical series of 30 cervico-mediastinal thyroid cancer patients operated on has been retrospectively reviewed. Results were compared with those obtained in patients operated on for benign cervico-mediastinal goiter and thyroid cancer confined to cervical region. Of 4688 thyroidectomies performed, 30 patients were operated on for thyroid carcinoma with cervico-mediastinal extension. There were 15 males and 15 females. The mean age was 67 years (range, 21-86 years). Patients with cervico-mediastinal cancer were significantly older than patients with benign cervico-mediastinal goiter (P < 0.0001). Time between onset of first symptoms and surgery was significantly longer in patients with cervico mediastinal cancer than in those with benign cervico-mediastinal goiter (P < 0.0001) and cervical thyroid cancer. Signs and symptoms at the time of surgery were cervical mass in 28 patients (93%), cervical lymphadenopathy in 20 patients (66%), dyspnea in 21 (70%), dysphagia in 9 (30%), dysphonia in 2 (7%), and venous stasis in 1 (3%). None of the patients was asymptomatic. Total thyroidectomy with functional lymphectomy was performed in 16 cases. Seven of these patients were operated on in 2 stages. In 8 cases the operation was a debulking procedure, and in 6 it was a near-total thyroidectomy. Sternotomy was performed in two cases. A differentiated thyroid cancer was found in 21 patients (70%), medullary in 5 (17%) and undifferentiated in 4 (13%). The incidence of medullary carcinoma was significantly higher compared with cervical cancer (P < 0.008). Postoperative complications were higher than those occurring in benign cervico-mediastinal goiter and similar to those occurring in cervical cancer. The actuarial survival was similar to that of cervical cancer matched for age and sex. This analysis shows that the longer clinical history of goiter is related to its endothoracic development and its neoplastic transformation. This finding should further encourage surgeons to treat any cervico-mediastinal goiter as promptly as possible. PMID- 10824752 TI - Effect of magnetic stimulation on the contractile activity of the rectum in humans. AB - Magnetic stimulation (MS) has been used to activate the neuromuscular tissue by inducing an electric field. Based on the results of a recent study on a canine model (Eur Surg Res 1998;30:268-72), which demonstrated that sacral MS effected a rectal and vesical pressure rise and a drop of rectal neck (anal canal) pressure, thereby achieving evacuation, the test was performed on 28 healthy volunteers (mean age, 36.6 years; 18 men and 10 women). The rectal, rectal neck, and vesical pressures were recorded during sacral MS with a magnetic coil while the rectum was empty and distended by a balloon. Electromyographic activity of the two rectus abdominis muscles was determined to exclude the possible interference of intra-abdominal pressure with the MS recordings. Stimulation parameters were set at 70 per cent intensity, 40-Hz frequency, and 1-to 2-second burst length. Sacral MS effected significant rectal and vesical pressure rise (P < 0.01 and P < 0.01, respectively) and drop of rectal neck pressure (P < 0.01). Intermittent stimulation induced balloon expulsion from the rectum. The two rectus abdominis muscles did not show change in electromyographic activity during MS, indicating that the rectal and vesical pressure rise was not due to increased intra abdominal pressure. Sacral MS induced rectal evacuation with no adverse effects. The method is simple, easy, safe, and noninvasive and is suggested to be applied for the treatment of the inertic or neuropathic rectum. As the vesical pressure proved to be elevated too, MS might also be used for rectal and vesical evacuation in patients with spinal cord lesions. PMID- 10824753 TI - Management of cystic neoplasms of the pancreas. AB - Cystic lesions of the pancreas are encountered frequently in clinical practice. Cystic neoplasms of the pancreas are much less common. In order to diagnose and most effectively treat these lesions, the surgeon must understand their clinical presentation in order to plan appropriate preoperative evaluation and therapeutic intervention. The following is a descriptive outline of the most common types of cystic neoplasms of the pancreas and a discussion of their preoperative, intraoperative, and postoperative management. The goal of the paper is to provide a functional understanding of cystic neoplasms of the pancreas so that they may be recognized and appropriately treated. PMID- 10824754 TI - Prognostic variables in male breast cancer. AB - The prognostic role of ploidy status, S phase fraction, estrogen and progesterone receptor status, and the expression of p53 and erbB-2 protein in male breast carcinoma (MBC) remains controversial. The primary objective of this study was to determine which of the common prognostic factors for female breast cancer predict prognosis in MBC. A secondary objective was to assess the impact of comorbid illnesses on survival. A retrospective review of demographic data, surgical treatment, pathological staging, adjuvant treatment and follow-up was completed for 16 patients with MBC (1 intraductal and 15 invasive). Formalin-fixed, paraffin-embedded tissue was processed for ploidy, S phase fraction, and immunohistochemical detection of estrogen and progesterone receptors plus expression of p53 and erbB-2 protein. Six of 15 patients with infiltrating ductal carcinoma are currently alive without evidence of disease and a median survival of 61 months. Nine patients died after a median survival of 52 months, with 6 patients having no evidence of recurrent breast cancer. Two of 3 deaths secondary to advanced breast cancer occurred in patients who initially presented with T4 lesions and were staged IIIB. Two of 15 tumors were erbB-2 positive, whereas only 1 tested weakly positive for p53 protein. We observed that MBCs express erbB-2 and p53 proteins infrequently. Neither ploidy status, S phase fraction, nor erbB 2/p53 status provided any apparent improvement in establishing prognosis beyond routine pathological staging. Advanced TNM stage was associated with diminished survival. The majority of MBCs express estrogen and progesterone receptors. Survivals in MBC were reduced in association with comorbid medical conditions. PMID- 10824755 TI - Re: Local excision and chemoradiation for low rectal T1 and T2 cancers is an effective treatment. PMID- 10824757 TI - Establishing benchmarks for ranking residencies. PMID- 10824756 TI - Re: Factors affecting the surgical management of infective endocarditis. PMID- 10824758 TI - Barriers to literature assessment during residency. PMID- 10824759 TI - Medical students' attitudes toward military medicine. PMID- 10824760 TI - Not much give left in the safety net. PMID- 10824761 TI - The AAMC and the CDC as strategic partners: why? And why now? Association of American Medical Colleges. PMID- 10824762 TI - Collaboration and peer review in medical schools' strategic planning. AB - The management of medical schools has never been so difficult, but can be facilitated by using a well-developed, broadly accepted strategic plan. While the concept of strategic planning has been reasonably well accepted by both faculty and leaders at most medical schools, using the strategic plan to allocate resources has proved to be a challenge. Achieving "buy-in" by all parties involved can help meet this challenge and can be critical to the success of strategic planning and management. The authors describe the collaborative planning process that the University of Wisconsin Medical School used to develop its 1998-2000 strategic plan. This unique effort culminated in using a peer review process--similar to that used by the National Institutes of Health (NIH)- and developing criteria to select a limited number of program priorities. This selection occurred after a school-wide process to solicit strategic program proposals had taken place. Over 130 faculty from most departments throughout the school helped develop and revise the strategic plan. There was frequent communication and engagement with faculty at all levels, which was important in gaining the faculty's acceptance and, indeed, endorsement of the process and its outcomes. Because the process was effective in achieving consensus about the school's strategic priorities, it enabled the school to reach a firmer end-point and implementation plan than had been possible with the previous strategic plan. It also identified important weaknesses in some areas of the medical school; the resulting attention to those areas will help strengthen the school. Finally, the process moved much more swiftly than the previous effort. The authors recommend that such an approach be used by other medical schools, and be carried out before a school implements mission-aligned budgeting and management of its fiscal resources. PMID- 10824763 TI - Measuring the costs of primary care education in the ambulatory setting. AB - In 1995, the authors obtained cost, operations, and educational activity data from 98 ambulatory care sites across the United States in which primary care teaching was occurring and compared those data with the corresponding data from 84 ambulatory care sites where no teaching was going on. The teaching sites in the sample were found to have 24-36% higher operating costs than the non-teaching sites. This overall difference in costs is approximately the same difference in costs earlier estimated for university teaching hospitals compared with non teaching hospitals. These costs are shared by all involved in the ambulatory education process: sponsors, sites, and faculty. In a related finding, the authors discovered that 30-50% of all ambulatory care sites thought not to be involved in education are in fact teaching at a high level of involvement. Further research into not only the costs but the value of education in the clinical setting is encouraged. The authors also hope that the publication of this report will encourage accrediting bodies and professional organizations to improve the information available about ambulatory care training in general. PMID- 10824764 TI - Review of the score-reporting policy for the United States Medical Licensing Examination. AB - In 1998, the authors, acting on behalf of the National Board of Medical Examiners (NBME), undertook a review of the scoring policy for the United States Medical Licensing Examination (USMLE). The main goal was to determine the likely effect of changing from numeric score reporting to reporting pass-fail status. Several groups were surveyed across the nation to learn how they felt they would be affected by such a change, and why: all 54 medical boards; 1,600 randomly selected examinees (including 250 foreign medical graduates) who had recently taken either Step 1, Step 2, or Step 3 of the USMLE; 2,000 residency directors; the deans, education deans, and student affairs deans at all 125 U.S. medical schools accredited by the Liaison Committee on Medical Education; and all 17 members of the Council of Medical Specialty Societies. Responses from the different groups surveyed varied from 80% to a little less than half. The authors describe in detail the various views of the respondents and their reasons. Some members in each group favored each of the reporting formats, but the trend was to favor numeric score reporting. The majority of the responding examinees desired that their USMLE scores be sent to them in numeric form but sent to their schools and to residency directors in pass-fail form. Based on the responses and a thorough discussion of their implications, the Composite Committee (which determines USMLE score-reporting policy) decided that there is no basis at this time for changing the current policy, but that it would review the policy in the future when necessary. PMID- 10824765 TI - The two fundamental duties of the physician. AB - The author asserts that physicians have two fundamental duties: they must balance the interests and wishes of the patient with the welfare of the health care system in which they practice. In fact, many physicians actually act in accordance with this dual approach, but they sense an inconsistency between their behaviors and their ideals, which focus on the patient only. The author explains why, as a consequence of this more complex role, medicine must stop viewing the case manager as the unavoidable but unwanted child within the family of medicine and must incorporate case management within the very identity of the physician. Finally, the author explains in detail that when the physician practices according to the dual-stewardship model just described, at least three beneficial consequences will emerge. First, the abysmal quality of end-of-life care in the United States may improve. Second, restraints on funding for last-chance or experimental treatments may, surprisingly, advance patients' interests. Third, an admission that much is not known about many mental illnesses and an acknowledgment that the money spent in treatment often yields limited results may encourage the health system to resist providing unproven treatments. This admission should also lead to more scientific knowledge and more effective approaches to these bewildering problems. In conclusion, the author states that it is not just the physician who must redefine himself in the new world of medicine. Administrators of managed care organizations must face the challenge of redirecting their traditional commitments to stockholders and boards of directors toward the patient-centered values of the physician. PMID- 10824766 TI - Balancing loyalties or splitting the difference? PMID- 10824768 TI - Medicine and the arts. The elephant man. PMID- 10824767 TI - Use of "emotional intelligence" as one measure of medical school applicants' noncognitive characteristics. PMID- 10824769 TI - The teaching of cultural issues in U.S. and Canadian medical schools. AB - PURPOSE: Despite the importance of culture in health care and the rapid growth of ethnic diversity in the United States and Canada, little is known about the teaching of cultural issues in medical schools. The study goals, therefore, were to determine the number of U.S. and Canadian medical schools that have courses on cultural issues, and to examine the format, content, and timing of those courses. METHOD: The authors contacted the deans of students and/ or directors of courses on cultural issues at all 126 U.S. and all 16 Canadian medical schools. Using a cross-sectional telephone survey, they asked whether each school had a course on cultural sensitivity or multicultural issues and, if so, whether it was separate or contained within a larger course, when in the curriculum the course was taught, and which ethnic groups the course addressed. RESULTS: The response rates were 94% for both U.S. (118) and Canadian (15) schools. Very few schools (U.S. = 8%; and Canada = 0%) had separate courses specifically addressing cultural issues. Schools in both countries usually addressed cultural issues in one to three lectures as part of larger, mostly preclinical courses. Significantly more Canadian than U.S. schools provided no instruction on cultural issues (27% versus 8%; p = .04). Few schools taught about the specific cultural issues of the largest minority groups in their geographic areas: only 28% and 26% of U.S. schools taught about African American and Latino issues, respectively, and only two thirds of Canadian schools taught about either Asian or Native Canadian issues. Only 35% of U.S. schools addressed the cultural issues of the largest minority groups in their particular states. CONCLUSIONS: Most U.S. and Canadian medical schools provide inadequate instruction about cultural issues, especially the specific cultural aspects of large minority groups. PMID- 10824770 TI - Measuring emotional intelligence of medical school applicants. AB - PURPOSE: To discuss the development, pilot testing, and analysis of a 34-item semantic differential instrument for measuring medical school applicants' emotional intelligence (the EI instrument). METHOD: The authors analyzed data from the admission interviews of 147 1997 applicants to a six-year BS/MD program that is composed of three consortium universities. They compared the applicants' scores on traditional admission criteria (e.g., GPA and traditional interview assessments) with their scores on the EI instrument (which comprised five dimensions of emotional intelligence), breaking the data out by consortium university (each of which has its own educational ethos) and gender. They assessed the EI instrument's reliability and validity for assessing noncognitive personal and interpersonal qualities of medical school applicants. RESULTS: The five dimensions of emotional intelligence (maturity, compassion, morality, sociability, and calm disposition) indicated fair to excellent internal consistency: reliability coefficients were .66 to .95. Emotional intelligence as measured by the instrument was related to both being female and matriculating at the consortium university that has an educational ethos that values the social sciences and humanities. CONCLUSION: Based on this pilot study, the 34-item EI instrument demonstrates the ability to measure attributes that indicate desirable personal and interpersonal skills in medical school applicants. PMID- 10824771 TI - Impact of a program to diminish gender insensitivity and sexual harassment at a medical school. AB - PURPOSE: To measure the effect of an intervention to reduce gender insensitivity and sexual harassment at one medical school. METHOD: Stanford University School of Medicine undertook a multifaceted program to educate faculty and students regarding gender issues and to diminish sexual harassment. The authors developed a survey instrument to assess the faculty's perceptions regarding environment (five scales) and incidences of sexual harassment. Faculty were surveyed twice during the interventions (1994 and 1995). RESULTS: Between the two years, the authors measured significant improvements in mean ratings for positive climate (p = .004) and cohesion (p = .006) and decreases in the faculty's perceptions of sexual harassment (p = 0006), gender insensitivity (p = .001), and gender discrimination (p = .004). The faculty also reported fewer observations of harassing behavior during the study period. CONCLUSIONS: An intervention program to diminish gender insensitivity and sexual harassment can measurably improve a medical school's environment. PMID- 10824772 TI - Long-term outcomes of the New Pathway Program at Harvard Medical School: a randomized controlled trial. AB - PURPOSE: To evaluate the long-term effects of an innovative curriculum, the New Pathway (NP) Program, on behaviors and attitudes related to humanistic medicine, lifelong learning, and social learning. METHOD: Long-term follow-up of Harvard Medical School students who participated in a randomized controlled trial. Descriptive study using 1998 telephone interviews of 100 1989 and 1990 graduates (50 who had studied the NP curriculum, 50 who had studied the traditional curriculum). The NP Program consisted of problem-based learning tutorials, with coordinated lectures, labs, experiences in humanistic medicine, and clinical experiences; the traditional program consisted of basic science lectures and labs. RESULTS: Of 22 measures on the survey, NP and traditional students differed significantly on only five (three humanism; two social learning): 40% of NP students and 18% of traditional students went on to practice primary care or psychiatry. NP students rated their preparation to practice humanistic medicine higher than did traditional students and expressed more confidence in their ability to manage patients with psychosocial problems. NP students were more likely than were traditional students to believe that faculty from the first two years continued to influence their thinking. NP students liked the pedagogic approaches of their program more than traditional students did. There was no difference between the groups on measures of lifelong learning. CONCLUSIONS: Differences between NP and traditional students in the humanism domain first appeared during medical school and residency and remained significant well into practice, suggesting that humanistic medicine can be taught and learned. PMID- 10824773 TI - Assessing the acquisition of core clinical skills through the use of serial standardized patient assessments. AB - PURPOSE: Students in many medical schools now undergo multiple standardized patient-based assessments. In this study, the authors examine the ability of such serial assessments to detect interval learning. METHOD: Twenty-one students from the University of California, Irvine, College of Medicine, class of 1999, underwent a clinical skills appraisal after three months of their third-year instruction. After nine months, all 89 members of that class completed an OSCE. Subsequently, all 87 students in the class of 2000 also completed clinical skills assessments after their third and ninth months of third-year instruction. All of these exercises included identical or similar stations measuring history, physical examination, and communication skills. Communication skills were measured somewhat differently during some of the exercises, using checklists that were either "content-" or "process-"oriented. The authors compared the performances for all groups. RESULTS: Both classes demonstrated significant improvement in physical examination performance, while their history performances remained unchanged. According to the assessments, their communication skills deteriorated over the course of their third-year instruction. Repeated exposures to similar or identical cases on the serial assessments did not impact the students' performances. Both content- and process-oriented measures of communication skills yielded highly similar results. CONCLUSIONS: Serial assessments using standardized patients can detect interval changes in performance that are independent of repeated exposures to similar or identical cases. Changes detected using this approach may have important curricular implications. PMID- 10824774 TI - Information needs and information seeking in community medical education. AB - PURPOSE: To investigate information needs and information seeking in primary care practices serving as educational sites. METHOD: The authors interviewed 15 community-based primary care physician-preceptors, once when the preceptor was working without a student and once when a student was present at the practice (in which case, the student was interviewed as well). The interview asked for questions that had arisen during the patient encounter or teaching moment that would need further investigation. A week after the interview, the authors contacted the preceptors to see whether they had pursued information to answer those questions. RESULTS: The preceptors generated fewer questions when students were present (0.42 versus 0.29 per patient). Both preceptors and students most frequently had questions pertaining to diagnoses and drug therapy. The proportion of questions pursued by the preceptors decreased when students were present (32% versus 16%). CONCLUSION: These findings cast light on information needs and seeking in the context of community medical education. Further research is needed to explore variables such as practice size and access to appropriate Web-based information resources. PMID- 10824775 TI - Educating residents about managed care: a partnership between an academic medical center and a managed care organization. AB - Physicians are taught to think in terms of individual patients rather than in terms of the health of a population, a view typically reinforced by residency training. Managers of managed care organizations estimate that it takes between one and two years of additional post-residency experience to prepare graduates of U.S. residency programs to practice in managed care settings. The authors describe a two-week block rotation in managed care and health system change that is required of all third-year medical residents at New York-Presbyterian Hospital/ Weill Cornell Medical Center. The program was developed through a partnership between the Joan and Sanford I. Weill Medical College of Cornell University, the New York-Presbyterian Hospital, and Empire BlueCross BlueShield. The authors discuss the rationale, curriculum content, teaching methods, and evaluation of the program, which is designed not only to educate residents about managed care but also to enable them to think critically about the changing health care system. The results of the program have exceeded expectations. Residents' knowledge of health economics, managed care concepts, and health system change, as measured by pre- and post-rotation tests, has shown a steep learning curve. Further, the residents have consistently given the rotation the highest ratings, and some residents have changed their career plans as a result of their participation. The program, which continues to expand, has demonstrated the potential of collaboration between traditional adversaries, an academic medical center and a managed care insurance company, and provides a replicable model for similar partnerships. PMID- 10824776 TI - A course for teaching patient-centered medicine to family medicine residents. AB - In 1988 the Department of Family Medicine at Ben Gurion University of the Negev in Israel developed a course that helps residents to acquire the attitudes and skills required for practicing patient-centered medicine. In the patient-centered approach, the physician relates to patients according to their needs rather than the doctor's own agenda, moving from professional control to patient empowerment. Though there are many elements to this method, certain basic orientations and skills are essential and must be taught, modeled, and reinforced in trainees. To accomplish these aims, a three-year course was developed, which is largely based on directed reading, open discussion, case presentations, role-plays, and Balint groups. It is composed of four levels, each of which must be mastered before residents can move to the next. The levels are (1) doctor-patient communication; (2) family-systems theory-concepts; (3) family-systems theory-practical applications; and (4) multidimensional approaches to simulated patients. In this article, the authors describe the course's concepts and content, and some indicators as to its influence on graduates. PMID- 10824777 TI - A theory-based faculty development program for clinician-educators. AB - This essay describes the development, implementation, and evaluation of a theory based faculty development program for physician-educators in medicine and pediatrics at The Cleveland Clinic. The program comprises a 12-hour course (focused on skills in precepting, bedside teaching, leading small-group discussions, giving lectures, designing curricula, and giving effective feedback); onsite coaching of teaching (on wards, in outpatient clinics, or in formal lectures); and innovative projects in clinical medical education. The program advocates and demonstrates a tailored-teaching approach combined with learner-centered, interactive, experiential, and reflective teaching strategies. The evaluation of the program includes satisfaction ratings, self-assessment of teaching competencies, and independent ratings of teaching effectiveness by the participants' trainees (medical students, residents, and fellows). The program is rated highly, shows significant improvements in teaching skills as measured by both participants' self assessments and independent ratings by participants' trainees, and is ongoing. PMID- 10824778 TI - Creating a culture of professionalism: an integrated approach. PMID- 10824779 TI - Promoting and assessing professionalism in the first two years of medical school. PMID- 10824780 TI - Self-accountability in medical student education. PMID- 10824781 TI - Medical school matriculants' perceptions of physician competencies and qualities. PMID- 10824782 TI - Assessing and changing unprofessional behaviors among faculty, residents, and students. PMID- 10824783 TI - Setting the culture of medical school from the start: appreciating and affirming diversity. PMID- 10824784 TI - Presenting cultural diversity and spirituality to first-year medical students. PMID- 10824785 TI - Integrating the care of Hispanic patients into a third-year clerkship. PMID- 10824786 TI - Enhancing residents' cultural competence through a lesbian and gay health curriculum. PMID- 10824787 TI - Anticipation of predictable stressors: a course to promote well-being for women physicians of color. PMID- 10824788 TI - Raising sensitivity to psychosocial issues of older adults using nutrition as a focus. PMID- 10824789 TI - Use of streaming video in preclinical lectures. PMID- 10824790 TI - Increasing the value of small-group learning. PMID- 10824791 TI - The hidden curriculum: a new undergraduate curriculum to evaluate students' perceptions of their third-year experiences. PMID- 10824792 TI - A course in end-of-life care for third-year medical students. PMID- 10824793 TI - Using a clerkship "mini-course" to gain student "buy-in" for learning preventive medicine. PMID- 10824794 TI - Changing teaching for changing times: the effect of a hospitalist program on the education of students. PMID- 10824795 TI - The human patient simulator: acceptance and efficacy as a teaching tool for students. The Medical Readiness Trainer Team. PMID- 10824796 TI - Problem-based learning in the medicine clerkship: a resident-driven strategy for teaching third-year students. PMID- 10824797 TI - A scholarly research requirement for medical students: the ultimate problem-based learning experience. PMID- 10824798 TI - Enhancing feedback to students using the mini-CEX (Clinical Evaluation Exercise). PMID- 10824799 TI - Assessment of competency in positioning and movement of physically disabled patients. PMID- 10824800 TI - A dissemination model for teaching evidence-based medicine. PMID- 10824801 TI - A medical school-managed care partnership to teach evidence-based medicine. PMID- 10824802 TI - The "EBM Rx": an initial experience with an evidence-based learning prescription. PMID- 10824803 TI - An evidence-based medicine curriculum for medical students: the art of asking focused clinical questions. PMID- 10824804 TI - An evidence-based physical diagnosis curriculum for third-year internal medicine clerks. PMID- 10824805 TI - The family competency project. PMID- 10824806 TI - Expanding the "standardized family" across three clerkships: a model for creating an interdisciplinary core curriculum in primary care. PMID- 10824807 TI - Partnership between a medical school and a managed care organization to financially support community-based teaching. PMID- 10824808 TI - First-year medical students designing a patient education handout. PMID- 10824809 TI - Incorporating the community in a community-based education program. PMID- 10824810 TI - Improved student learning and community health: the CCPH faculty service-learning institute. PMID- 10824811 TI - Medical students as ambassadors to Wisconsin communities. PMID- 10824812 TI - A four-year, longitudinal introduction to clinical medicine course organized in week-long blocks. PMID- 10824813 TI - A longitudinal community-based ambulatory curriculum using community preceptor resident-student teams. PMID- 10824814 TI - A university partnership for longitudinal ambulatory care education. PMID- 10824816 TI - MedFiler: a user-driven, decentralized database model for online medical education. PMID- 10824815 TI - Reducing reliance on hospitalized patients for undergraduate clinical skills teaching in internal medicine. PMID- 10824817 TI - Use of web-based technology in a peer-teaching program. PMID- 10824818 TI - Web-based peer evaluation by medical students. PMID- 10824819 TI - Using web-based case presentations to supplement a surgery clerkship curriculum. PMID- 10824820 TI - Development and formative evaluation of a longitudinal web-based nutrition curriculum. PMID- 10824821 TI - Using PalmPilots as a teaching tool during a primary care clerkship. Advanced Education Group. PMID- 10824822 TI - Factors influencing internal medicine program directors' decisions about applicants. PMID- 10824823 TI - Objectives-based self-assessment of surgical residents. PMID- 10824824 TI - Prisons: learning about women's health and substance abuse. PMID- 10824825 TI - Clinic without walls: exploring community resources in an internal medicine residency. PMID- 10824826 TI - A multidisciplinary approach to teaching residents to teach. PMID- 10824827 TI - Applying adult learning theory to a residents-as-teachers workshop series. PMID- 10824828 TI - Development of a web-based GME core curriculum. PMID- 10824829 TI - GME core curriculum: a pilot program in radiology. Graduate medical education. PMID- 10824830 TI - Primary care chief residency. PMID- 10824831 TI - Teaching residents to develop healing relationships when caring for the chronically ill. PMID- 10824832 TI - Medical residency training in the management of obesity. PMID- 10824833 TI - Trauma videotape resuscitation conference: a new way to teach trauma care. PMID- 10824834 TI - Patient safety curriculum. PMID- 10824835 TI - A systematic approach to teaching insertion of a central venous line. PMID- 10824836 TI - A cardiac physical examination curriculum. PMID- 10824837 TI - The Champions Project: a two-tiered mentoring approach to faculty development. PMID- 10824838 TI - A faculty development workshop on "developing successful workshops". PMID- 10824839 TI - Writing "blitzes" for medical educators. PMID- 10824840 TI - A computer "boot camp" for academic medicine faculty. PMID- 10824841 TI - Feedback notes: a system for feedback to students and residents. Advanced Education Faculty Development Group. PMID- 10824842 TI - A workshop for junior faculty on the learner in difficulty. PMID- 10824843 TI - Chancellor's Educational Retreat for interdisciplinary faculty development in women's health. PMID- 10824844 TI - Using residents' ratings of teaching to assess the effectiveness of faculty development. PMID- 10824845 TI - The faculty self-efficacy scale: a tool for evaluating faculty development interventions. PMID- 10824846 TI - Evening report. PMID- 10824847 TI - Using an OSCE to assess primary care physicians' competence in breaking bad news. PMID- 10824848 TI - Behavioral medicine teaching rounds: improving doctor-patient interaction skills. PMID- 10824849 TI - A collaborative practice-based learning model for residents and faculty. PMID- 10824850 TI - The populations and quality improvement seminar for medical residents. PMID- 10824851 TI - Supramolecularity creates nonstandard protein ligands. AB - Congo red and a group of structurally related dyes long used to stain amyloid proteins are known to associate in water solutions. The self-association of some dyes belonging to this group appears particularly strong. In water solutions their molecules are arranged in ribbon-like micellar forms with liquid crystalline properties. These compounds have recently been found to form complexes with some native proteins in a non-standard way. Gaps formed by the local distribution of beta-sheets in proteins probably represent the receptor sites for these dye ligands. They may result from higher structural instability in unfolding conditions, but also may appear as long range cooperative fluctuations generated by ligand binding. Immunoglobulins G were chosen as model binding proteins to check the mechanism of binding of these dyes. The sites of structural changes generated by antigen binding in antibodies, believed to act as a signal propagated to distant parts of the molecule, were assumed to be suitable sites for the complexation of liquid-crystalline dyes. This assumption was confirmed by proving that antibodies engaged in immune complexation really do bind these dyes; as expected, this binding affects their function by significantly enhancing antigen binding and simultaneously inhibiting C1q attachment. Binding of these supramolecular dyes by some other native proteins including serpins and their natural complexes was also shown. The strict dependence of the ligation properties on strong self-assembling and the particular arrangement of dye molecules indicate that supramolecularity is the feature that creates non-standard protein ligands, with potential uses in medicine and experimental science. PMID- 10824852 TI - Plasmid-mediated suppression of the mutational activation of the bgl operon in Shigella sonnei. AB - SSOR, a clinical isolate of Shigella sonnei which exhibits a Salicin-negative phenotype, is unable to mutate to give rise to Sal+ derivatives although a homolog of the Escherichia coli bgl operon is retained by the strain. This was correlated to the presence of an endogenous plasmid in the strain. A plasmid cured derivative, AK711, could give rise to Sal+ mutants in two steps. Introduction of the plasmid DNA, extracted from SSOR, into various strains of E. coli and S. sonnei, resulted in ampicillin resistant transformants. Interestingly, the presence of the plasmid suppressed the mutational activation of the bgl operon in the transformants. This was further substantiated by the observation that, transformants that have lost the plasmid regained the ability for mutational activation of the bgl operon. Preliminary characterisation of the plasmid indicated a size of 3.8 kb with an origin of replication resembling that of ColE1 replicons and the bla gene homolog of Tn3. Observations of the mutation frequency at the srl and lac loci in the presence of the plasmid indicate that there is a reduction in the mutation frequency, suggesting an antimutator activity associated with the plasmid. PMID- 10824853 TI - Involvement of the essential yeast DNA polymerases in induced gene conversion. AB - In the yeast Saccharomyces cerevisiae three different DNA polymerases alpha, delta and epsilon are involved in DNA replication. DNA polymerase alpha is responsible for initiation of DNA synthesis and polymerases delta and epsilon are required for elongation of DNA strand during replication. DNA polymerases delta and epsilon are also involved in DNA repair. In this work we studied the role of these three DNA polymerases in the process of recombinational synthesis. Using thermo-sensitive heteroallelic mutants in genes encoding DNA polymerases we studied their role in the process of induced gene conversion. Mutant strains were treated with mutagens, incubated under permissive or restrictive conditions and the numbers of convertants obtained were compared. A very high difference in the number of convertants between restrictive and permissive conditions was observed for polymerases alpha and delta, which suggests that these two polymerases play an important role in DNA synthesis during mitotic gene conversion. Marginal dependence of gene conversion on the activity of polymerase epsilon indicates that this DNA polymerase may be involved in this process but rather as an auxiliary enzyme. PMID- 10824854 TI - Fragments of LINE-1 retrotransposons flanked by inverted telomeric repeats are present in the bovine genome. Homology with human LINE-1 elements. AB - In the bovine genome we found two intrachromosomal DNA fragments flanked by inverted telomeric repeats (GenBank Accession Nos. AF136741 and AF136742). The internal parts of the fragments are homologous exclusively to the human sequences and to the consensus sequence of the L1MC4 subfamily of LINE-1 retrotransposons which are widespread among mammalian genomes. We found that distribution of homologous human sequences within our fragments is not random, reflecting a complicated pattern of insertion mechanisms of and maintenance of retrotransposons in mammalian genomes. One of the possible explanations of the origin of LINE-1 truncated elements flanked by inverted telomeric repeats in the bovine genome is that extrachromosomal DNA fragments may be modified by telomerase and subsequently, transferred into chromosomal DNA. PMID- 10824855 TI - Translational frameshift sites within bacteriophage lambda genes rexA and cI. AB - Phage lambda's cI-rexA-rexB operon displays an intriguing example of regulation by an unexplained mechanism of polarity. We have identified three potential -1 translational frameshift sites and present a model for translational frameshift suppression by lambda's CI repressor as a mechanism of regulating operon polarity, implying an additional role for CI self-regulation. PMID- 10824856 TI - Cloning of the lymphoid enhancer binding factor-1 (Lef-1) cDNA from rat kidney: homology to the mouse sequence. AB - We have cloned and sequenced rat cDNA that encodes the Lef-1 protein. The cDNA, containing 1194 nt exhibits 94% similarity to the mouse Lef-1 cDNA. The deduced amino-acids sequence of rat Lef-1 protein, consisting of 397 amino acids, exhibited 98% homology with the known sequence of mouse Lef-1 protein. PMID- 10824858 TI - Extraribosomal function of the acidic ribosomal P1-protein YP1alpha from Saccharomyces cerevisiae. AB - The yeast acidic ribosomal P-proteins YP1alpha, YP1beta, YP2alpha and YP2beta were studied for a possible transactivation potential beside their ribosomal function. The fusions of P-proteins with the GAL4 DNA-binding domain were assayed toward their transcriptional activity with the aid of reporter genes in yeast. Two of the P-proteins, YP1alpha and YP1beta, exhibited transactivation potential, however, only YP1alpha can be regarded as a potent transactivator. This protein was able to transactivate a reporter gene associated with two distinct promoter systems, GAL1 or CYC1. Additionally, truncated proteins of YP1alpha and YP1beta were analyzed. The N-terminal part of YP1alpha fused to GAL4-BD showed transactivation potential but the C-terminal part did not. Our results suggest a putative extraribosomal function for these ribosomal proteins which consequently may be classified as "moonlighting" proteins. PMID- 10824857 TI - Probing iso-1-cytochrome c structure by site-directed spin labeling and electron paramagnetic resonance techniques. AB - A cysteine-specific methanethiosulfonate spin label was introduced into yeast iso 1-cytochrome c at three different positions. The modified forms of cytochrome c included: the wild-type protein labeled at naturally occurring C102, and two mutated proteins, S47C and L85C, labeled at positions 47 and 85, respectively (both S47C and L85C derived from the protein in which C102 had been replaced by threonine). All three spin-labeled protein derivatives were characterized using electron paramagnetic resonance (EPR) techniques. The continuous wave (CW) EPR spectrum of spin label attached to L85C differed from those recorded for spin label attached to C102 or S47C, indicating that spin label at position 85 was more immobilized and exhibited more complex tumbling than spin label at two other positions. The temperature dependence of the CW EPR spectra and CW EPR power saturation revealed further differences of spin-labeled L85C. The results were discussed in terms of application of the site-directed spin labeling technique in probing the local dynamic structure of iso-1-cytochrome c. PMID- 10824859 TI - SDS/PAGE characteristics of protein kinases tightly associated with chick embryo brain ribosomes. AB - Protein kinases tightly associated with chick embryo brain ribosomes washed with Triton X-100 and KCl were characterized by their ability to phosphorylate ribosomes and two exogenous substrates, histone IIA and casein. c-AMP-dependent kinase (PKA) and casein kinases (CK1, CK2) were examined in the presence of specific modulators by SDS/PAGE followed by renaturation in gel assay according to Kameshita & Fujisawa (Anal. Biochem. 1989, 183, 139-143). Basing on these data it can be presumed that PKA activity increases, but the levels of CK2 and CK1 decrease during chick embryo development. PMID- 10824860 TI - Kinetic studies on the oxidation of nitrite by horseradish peroxidase and lactoperoxidase. AB - The reaction of nitrite (NO2-) with horseradish peroxidase and lactoperoxidase was studied. Sequential mixing stopped-flow measurements gave the following values for the rate constants of the reaction of nitrite with compounds II (oxoferryl heme intermediates) of horseradish peroxidase and lactoperoxidase at pH 7.0, 13.3 +/- 0.07 mol(-1) dm3 s(-1) and 3.5 +/- 0.05 x 10(4) mol(-1) dm3 s( 1), respectively. Nitrite, at neutral pH, influenced measurements of activity of lactoperoxidase with typical substrates like 2,2'-azino-bis[ethyl-benzothiazoline (6)-sulphonic acid] (ABTS), guaiacol or thiocyanate (SCN-). The rate of ABTS and guaiacol oxidation increased linearly with nitrite concentration up to 2.5-5 mmol dm(-3). On the other hand, two-electron SCN- oxidation was inhibited in the presence of nitrite. Thus, nitrite competed with the investigated substrates of lactoperoxidase. The intermediate, most probably nitrogen dioxide (*NO2), reacted more rapidly with ABTS or guaiacol than did lactoperoxidase compound II. It did not, however, effectively oxidize SCN- to OSCN-. NO2- did not influence the activity measurements of horseradish peroxidase by ABTS or guaiacol method. PMID- 10824861 TI - Low molecular mass products of depolymerization of purified mucin--attempts at isolation and characterization. AB - Samples of crude mucin were incubated at room temperature for 48 and 96 h in a sodium azide containing buffer, pH 7.0. Then each sample was purified, reduced and alkylated with iodo[14C]acetamide. Electrophoretic analysis demonstrated that radioactivity was incorporated into the mucin subunits and proteins of 100 and 140 kDa. The results of our experiments suggest that the released proteins can be a part of mucin molecule, cleaved by proteolysis and reduction of disulfide bridges. PMID- 10824862 TI - Heparin- and Zn2+-binding proteins from boar seminal plasma. AB - Low molecular mass, heparin-binding proteins from seminal plasma play an important role in gametes interaction whereas plasmatic Zn2+-binding proteins stabilize chromatin and plasmalemma structures and protect spermatozoa in the female reproductive tract. By means of affinity chromatography the heparin- and Zn2+-binding proteins were isolated from boar seminal plasma and both preparations were analyzed by reverse HPLC. Most of the proteins bound to heparine and Zn2+-ions were classified as spermadhesins. Three fractions binding exclusively Zn2+ were isolated. They differ in amino-acid composition, content of glucosamine and content of protein components revealed by SDS/PAGE. PMID- 10824863 TI - Accumulation of collagen in ovarian benign tumours. AB - Extracellular matrix components of benign ovarian tumours (cystadenoma, adenofibroma, cystadenofibroma) were analysed. The investigated tumours contained twice as much collagen than control ovarian tissues. Significant alterations in mutual quantitative relationships between collagens of various types were observed. The proportion of type I collagen decreased and that of type III collagen increased. The accumulation of collagen was accompanied by a reduction in sulphated glycosaminoglycan content whereas the amount of hyaluronic acid was not changed. Dermatan sulphate was the most abundant glycosaminoglycan component. It is suggested that the accumulation of collagen (natural barrier to the migration of tumour cells) and underexpression of glycosaminoglycans/proteoglycans (binding some growth factors and interleukins) may exert an inhibitory effect on tumour growth. PMID- 10824864 TI - Identification of actin from hepatoma Morris 5123 cells. AB - The hepatoma Morris 5123 tumor growth is accompanied by changes in actin content and polymerization (Malicka-Blaszkiewicz et al. (1995) Mat. Med. Pol., 27, 115 118; Nowak et al. (1995) J. Exp. Cancer Res. 14, 37-40). Presently actin isoforms from cytosol and cytoskeleton fractions were separated by SDS/PAGE and identified with antibodies directed against different actin isoforms. Actin isolated from the cytosol by affinity chromatography on DNase I bound to agarose shows the presence of only one protein spot on 2D gel electrophoresis corresponding to the mobility of the rabbit a skeletal muscle actin (Mr 43,000) and isoelectric point equal to 5.3. It interacts only with monoclonal anti beta actin isoform antibodies, posing the question of differential affinity of actin isoforms to DNase I. PMID- 10824865 TI - Inhibitory effect of resveratrol on free radical generation in blood platelets. AB - Resveratrol (3,4',5-trihydroxystilbene), a compound found in many plants, has been shown to prevent coronary heart diseases and to exert a variety of antiinflammatory and anticancerogenic effects. It is effective in lowering the level of serum lipids and in inhibiting platelet aggregation. We evaluated the effect of trans-resveratrol on the production of free radicals in pig blood platelets and showed that resveratrol inhibited the production of different reactive oxygen species (O2*-, H2O2, singlet oxygen and organic radicals) measured by the luminol-dependent chemiluminescence in resting platelets (P < 0.05). Resveratrol inhibited also the generation of radicals in platelets activated by thrombin (P < 0.05). Treatment of platelets with resveratrol at concentrations of 6.25 and 12.5 microg/ml caused a statistically insignificant increase in the production of O2*- in these cells, as measured by reduction of cytochrome c; however, at higher doses (25, 50 and 100 microg/ml) resveratrol distinctly reduced the generation of O2*- in platelets (P < 0.05). We suggest that free radicals play an important role in the reduced reactivity of blood platelets induced by resveratrol. PMID- 10824866 TI - Inhibition of tumor growth by interleukin 10 gene transfer in B16(F10) melanoma cells. AB - Interleukin 10 (IL-10) is a potent immunosuppressive cytokine with an antitumor activity. The effect of IL-10 on tumor growth was tested in murine melanoma cells manipulated by gene transfer to secrete IL-10. In mice bearing B16(F10) tumors expressing IL-10 tumor growth was decreased depending on the amount of secreted IL-10. PMID- 10824867 TI - Protection of cattle against bovine leukemia virus (BLV) infection could be attained by DNA vaccination. AB - The bovine leukemia virus (BLV) envelope gene encoding extracellular glycoprotein gp51 and transmembrane glycoprotein gp30 was cloned into a vehicle expression vector under the human cytomegalovirus (CMV) intermediate early promoter. The intramuscular injection of this plasmid vector generated a cellular immune response. Seven out of ten cows vaccinated with the DNA construct resisted a drastic challenge (500 BLV-infected lymphocytes as an infectious dose). PMID- 10824868 TI - Isoenzymes of N-acetyl-beta-hexosaminidase in complicated pregnancy. AB - The activity of N-acetyl-beta-hexosaminidase was found to be significantly higher in the placentas collected after delivery from women in puerperium with symptoms of prolonged pregnancy or complicated by EPH gestosis, than in placentas from normal pregnancy. Isoelectrofocusing of placenta homogenates showed the presence of isoenzymes A, P and B of N-acetyl-beta-hexosaminidase. Different isoenzyme A patterns in homogenates were observed in placentas obtained from normal and prolonged pregnancies and in those complicated by EPH gestosis. PMID- 10824869 TI - Elevation of plasma fibrinogen in silent myocardial ischaemia. AB - High plasma levels of fibrinogen and plasminogen activator inhibitor (PAI-1) are reported to be correlated with coronary heart disease. Therefore the level of fibrinogen concentration in plasma was examined and verified for the possible correlation with the previously explored PAI-1 antigen and PAI-1 activity in the pathogenesis of premature atherosclerosis (Grzywacz et al., 1998, Blood Coagul Fibrinol. 9, 245-249). Examination included only men, aged 33-46 years, who were in a stable condition for at least six months after the acute event. They were divided into two subgroups: group A (n = 14) with and group B (n = 15) without ischaemic changes in 24 h Holter electrocardiogram. The number of involved vessels visible on the coronarography picture was similar in both groups. In the patients of group A the mean level of fibrinogen (3.92 vs 3.23 g/l, P < 0.05) was higher than in the controls (n = 15). No statistically differences were found between group B and control healthy subjects in any of the parameters measured. There were no correlation between fibrinogen concentration and PAI-1 antigen and activity levels, which were elevated in both groups of patients according to our previous study. Our results indicate that elevated levels of plasma fibrinogen and PAI-1 appeared in the group of patients with more severe disease, as revealed by silent myocardial ischaemia. PMID- 10824870 TI - The access of metabolites into yeast mitochondria in the presence and absence of the voltage dependent anion selective channel (YVDAC1). AB - Since yeast Saccharomyces cerevisiae mutants depleted of the voltage dependent anion selective channel (YVDAC1) are still able to grow on a non-fermentable carbon source, a functional transport system in the outer mitochondrial membrane must exist to support the access of metabolites into mitochondria. It was assumed that the properties of the system could be inferred from the differences in the results observed between wild type and mutant mitochondria since no crucial differences in this respect between the two types of mitoplasts were observed. YVDAC1-depleted mitochondria displayed a highly reduced permeability of the outer membrane, which was reflected in increased values of K0.5(NADH) for respiration and K0.5(ADP) for triggering phosphorylating state as well as in delayed action of carboxyatractylate (CATR) in inhibition of phosphorylating state. The parameters were chosen to express the accessibility of the applied species to the intermembrane space. The passage of the molecules through the outer membrane depleted of YVDAC1 could be partially improved in the presence of bivalent cations (Mg2+, Ca2+), as in their presence lower values of the calculated parameters were obtained. The restrictions imposed on the transport of molecules through the YVDAC1-depleted outer membrane resulted in a competition between them for the access to the intermembrane space as measured by changes in parameters observed for a given species in the presence of another one. The competition was stronger in the absence of Mg2+ and depended on charge and size of transported molecules, as the strongest competitor was CATR and the weakest one--NADH. Thus, it can be concluded that the transport system functioning in the absence of YVDAC1 is modulated by bivalent cations and charge as well as size of transported molecules. Since an increased level of respiration due to the dissipation of delta psi causes an increase of K0.5(NADH) in both wild type and YVDAC1-depleted mitochondria it is concluded that a common property of YVDAC1 and the system functioning in YVDAC1-depleted mitochondria seems to be the dependence of the capacity on the level of mitochondrial respiration. PMID- 10824871 TI - Chemical characterization of effective and ineffective strains of Rhizobium leguminosarum bv. viciae. AB - Chemical composition of lipopolysaccharide (LPS) isolated from an effective (97) and ineffective (87) strains of R. l. viciae has been determined. LPS preparations from the two strains contained: glucose, galactose, mannose, fucose, arabinose, heptose, glucosamine, galactosamine, quinovosamine, and 3-N-methyl-3,6 dideoxyhexose, as well as glucuronic, galacturonic and 3-deoxyoctulosonic acid. The following fatty acids were identified: 3-OH 14:0, 3-OH 15:0, 3-OH 16:0, 3-OH 18:0 and 27-OH 28:0. The ratio of 3-OH 14:0 to other major fatty acids in LPS 87 was higher that in LPS 97. SDS/PAGE profiles of LPS indicated that, in lipopolysaccharides, relative content of S form LPS I to that of lower molecular mass (LPS II) was much higher in the effective strain 97 than in 87. All types of polysaccharides exo-, capsular-, lipo, (EPS, CPS, LPS, respectively) examined possessed the ability to bind faba bean lectin. The degree of affinity of the host lectin to LPS 87 was half that to LPS 97. Fatty acids (FA) composition from bacteroids and peribacteroid membrane (PBM) was determined. Palmitic, stearic and hexadecenoic acids were common components found in both strains. There was a high content of unsaturated fatty acids in bacteroids as well as in PBM lipids. The unsaturation index in the PBM formed by strain 87 was lower than in the case of strain 97. Higher ratio of 16:0 to 18:1 fatty acids was characteristic for PMB of the ineffective strain. PMID- 10824872 TI - Gastroenterology in the Asian-Pacific region in the new millennium: the role of the Journal of Gastroenterology and Hepatology. PMID- 10824873 TI - Hepatitis B virus X protein: searching for a role in hepatocarcinogenesis. PMID- 10824874 TI - Lessons from animal models of primary biliary cirrhosis. PMID- 10824875 TI - Anti-neutrophil cytoplasmic antibodies in hepatobiliary disease. PMID- 10824876 TI - How to manage hepatic vein tumour thrombus in hepatocellular carcinoma. PMID- 10824877 TI - Impact of alcohol on the ability of Kupffer cells to produce chemokines and its role in alcoholic liver disease. AB - Chemokines are implicated in the pathogenesis of alcoholic liver disease in humans and in experimental models of alcohol intoxication. The major sources of these chemokines are Kupffer cells which represent more than 80% of tissue macrophages in the body. Kupffer cells are highly responsive to the effects of ethanol, endotoxin and human immunodeficiency virus (HIV)-1 glycoprotein120. These agents, either independently or in combination, may exacerbate the production of chemokines. Chemokines are agents that are highly chemotactic to mononuclear cells and granulocytes. The levels of these chemokines in sera and tissue are elevated in patients with alcoholic hepatitis, alcoholic cirrhosis, diseased livers, viral hepatitis, and in experimental models of chronic alcohol intoxication. Alcohol-induced influx of endotoxin from the gut into the portal circulation is suggested to play an important role in the activation of Kupffer cells which leads to enhanced chemokine release. The up-regulation of chemokines during alcohol consumption is selective. During the early phase of alcoholic liver disease, C-X-C or alpha-chemokines predominate. This is also associated with neutrophilic infiltration of the liver. In the later stage, up-regulation of C-C or beta-chemokine production and migration of mononuclear cells into the liver are observed, and this may lead to liver cirrhosis. Selective up-regulation of chemokine synthesis and release may involve differential modulation of the transcription factors required for chemokine gene expression. Increased cytokine release following alcohol consumption may also regulate chemokine secretion in Kupffer cells via paracrine and autocrine mechanisms and vice versa. In addition, infection with HIV-1 may further compromise the liver to more damage. During HIV 1 infection, a pre-existing liver disease superimposed on chronic alcohol consumption may also exacerbate HIV-1 replication and lymphocytic infiltration in the liver, because of the ability of HIV-1 gp120 to stimulate chemokine production by Kupffer cells and stimulate migration of inflammatory leucocytes in the liver. PMID- 10824878 TI - Putative role of hepatitis B virus X protein in hepatocarcinogenesis: effects on apoptosis, DNA repair, mitogen-activated protein kinase and JAK/STAT pathways. AB - Chronic infection with hepatitis B virus (HBV) is a major risk factor for the development of hepatocellular carcinoma (HCC). The pathogenesis of HBV-induced malignant transformation is, however, incompletely understood. HBx, the protein encoded by the X open reading frame, is a transcriptional activator that has been implicated in hepatocarcinogenesis. HBx inhibits the function of the tumour suppressor protein p53 in what is thought to be an early event in hepatocyte transformation before the later accumulation of inactivating p53 point mutations. HBx inhibits apoptosis but also exerts pro-apoptotic effects. The effects of HBx on apoptosis may be important not only for the development of HCC but also for the establishment of HBV infection. Further implication of HBx in hepatocyte transformation has been the demonstration that it inhibits the repair of damaged hepatocyte DNA. This effect may be mediated by interaction with p53 or through binding to the damaged DNA binding protein (DDB), which plays an accessory role in nucleotide excision repair. In addition, HBx activates cell signalling cascades involving mitogen-activated protein kinase (MAPK) and Janus family tyrosine kinases (JAK)/signal transducer and activators of transcription (STAT) pathways. The implications of these modulating effects of HBx are not fully understood, but they are likely to have wide-ranging effects on hepatocyte proliferation, apoptosis and the regulation of cell growth checkpoints. The cellular functions ascribed to HBx are unusually diverse, and defining the biologically important role of HBx during HBV replication will go some way to understanding the sequelae of chronic HBV infection. PMID- 10824879 TI - Rapid urease test is less sensitive than histology in diagnosing Helicobacter pylori infection in patients with non-variceal upper gastrointestinal bleeding. AB - BACKGROUND AND AIMS: The validity of the rapid urease (CLO) test to diagnose Helicobacter pylori infection in patients with bleeding ulcers has been questioned. The aim of this paper is to evaluate the validity of the CLO test in comparison with histology in diagnosing H. pylori infection in patients with acute upper gastrointestinal bleeding (UGB), irrespective of non-steroidal anti inflammatory drug (NSAID) use. METHODS: Upper gastrointestinal endoscopy was performed within 24 h of admission for all patients with UGB admitted to the Department of Pathophysiology, Medical School, Athens, for a period of 12 months. Patients with variceal bleeding, previous gastric operation, recent treatment with proton pump inhibitors (< 2 months) and those with a history of H. pylori eradication therapy were excluded from the study. At least four biopsies (two from the antrum and two from the body) were obtained for the CLO test and histology (modified Giemsa). RESULTS: Seventy-two consecutive patients (aged 18 90 years, 51 men, 21 women) were included. Forty-six patients (64%) used NSAID. Thirty-two patients (44%) were found to be positive for H. pylori infection by the CLO test, while 44 patients (61%) were found to be positive on histology (P<0.045, 95% CI, 0.004-0.331). The sensitivity and specificity of the CLO test were 68 and 93% respectively; positive and negative predictive values were 94 and 65%, respectively. The age of the patient and visible blood in the stomach did not influence results of either the CLO or histology. CONCLUSIONS: The CLO test, performed within 24 h of hospital admission in patients with UGB, irrespective of NSAID use, is unreliable for the detection of H. pylori infection. The age of the patient and the presence of blood in the stomach do not seem to influence these results. PMID- 10824880 TI - Plaunotol suppresses interleukin-8 secretion induced by Helicobacter pylori: therapeutic effect of plaunotol on H. pylori infection. AB - BACKGROUND: It has been suggested that gastric mucosal injury induced by Helicobacter pylori infection is mediated by interleukin-8 (IL-8). METHODS: We studied the effect of plaunotol, a drug extracted from the Plau-noi tree of Thailand, and reported it to be effective in the treatment of ulcers, of IL-8 secretion induced by H. pylori and of the inhibitory adhesion activity of the bacterium to gastric epithelial cells. Moreover, the therapeutic effect of plaunotol on H. pylori infection was assessed by using the gnotobiotic murine model. RESULTS: Plaunotol inhibited the growth of H. pylori (1.5 x 10(4) c.f.u./mL) at high doses (24-48 microg/mL), but not at low doses (3-6 microg/mL). Interleukin-8 secretion induced by H. pylori was inhibited by coculture with plaunotol in a dose-dependent manner. The adhesion of H. pylori to MKN45 cells was also suppressed by coculture with plaunotol in a dose-dependent manner. An in vivo study showed that plaunotol improved histological gastritis and decreased the H. pylori antibody titre. CONCLUSIONS: These findings suggest that plaunotol has a therapeutic effect on gastritis induced by H. pylori. PMID- 10824881 TI - Evaluation of gut motility in type II diabetes by the radiopaque marker method. AB - BACKGROUND: The clinical usefulness of the radiopaque marker method for detecting diabetic gastrointestinal motility disturbances, was evaluated by examining 21 type II diabetes subjects who did not have any neuropathic symptoms. METHODS: After administration of a Sitzmark capsule, markers were located using plain abdominal radiographs, and the transit time of the markers through seven areas of digestive tract was calculated by Arhan's methods. The plasma concentration of acetaminophen at 45 min after oral administration was measured to evaluate gastric emptying time. The coefficient of variation of R-R intervals on the electrocardiograms (CV(R-R)) was measured to evaluate parasympathetic autonomic function. RESULTS: In the diabetics, the average (+/- SD) transit time through upper digestive tracts was slightly but not significantly elongated compared with control subjects (14.4 +/- 8.3 vs 9.9 +/- 6.1 h). Significant elongation was observed in transit time through the lower digestive tracts or the whole gut (44.6 +/- 20.9 and 57.9 +/- 22.3 h, respectively) compared with control subjects (23.3 +/- 8.5 and 33.2 +/- 11.0 h). The transit time of the markers from stomach to small intestine was highly correlated (r = 0.693) with plasma concentration of acetaminophen. The transit time through either the whole colon (r = 0.564) or the whole gut (r = 0.630) was highly correlated with CV(R-R). CONCLUSIONS: These findings suggest that the radiopaque marker method is a useful tool for detecting the sections of the digestive tract responsible for gut motility disturbances. In type II diabetics with no neuropathic symptoms, the lower digestive tracts may deteriorate prior to the impairment of upper digestive tracts. PMID- 10824882 TI - Aspartate aminotransferase: alanine aminotransferase ratio in chronic hepatitis C infection: is it a useful predictor of cirrhosis? AB - BACKGROUND: The clinical usefulness of the ratio of serum aspartate aminotransferase (AST) to alanine aminotransferase (ALT) has been explored in several liver disorders. It has been suggested that in patients with chronic hepatitis C virus (HCV) infection an AST:ALT > or = 1 has 100% specificity and positive predictive value in distinguishing cirrhotic from non-cirrhotic patients. Such statistical certainty attached to a simple biochemical test merits further evaluation. The present study, therefore, assessed the AST:ALT in patients with chronic HCV infection to determine the validity of the ratio in predicting cirrhosis and to correlate the ratio with the histological grade of necroinflammatory activity and fibrosis. METHODS: A retrospective analysis of 153 patients with chronic HCV infection was conducted. Serum biochemistry had been obtained within a mean of 4 weeks of liver biopsy. The histology was scored in terms of activity and fibrosis as described by Scheuer and correlated with AST:ALT. RESULTS: In 30 patients with cirrhosis, the mean AST:ALT (0.99 +/- 0.06) was higher than in 123 patients without cirrhosis (0.60 +/- 0.02; P < 0.001). A ratio > or = 1 had 95.9% specificity and 73.7% positive predictive value in distinguishing cirrhotic from non-cirrhotic patients, with a 46.7% sensitivity and 88.1% negative predictive value. The ratio also parallelled the Scheuer score with respect to fibrosis but not with respect to inflammation. CONCLUSION: Although relatively insensitive, an AST:ALT > or = 1 is highly specific but not diagnostic for the presence of cirrhosis in patients with chronic HCV infection. The ratio reflects the grade of fibrosis in these patients. PMID- 10824883 TI - Low incidence of hepatitis C virus transmission between spouses: a prospective study. AB - BACKGROUND: Interspousal transmission of hepatitis C virus (HCV) has been documented; however, the annual risk of interspousal transmission remains unclear. METHODS: A long-term prospective study to define the risk of interspousal transmission of HCV was conducted. One hundred and twelve index patients with chronic hepatitis C and their anti-HCV seronegative spouses were enrolled. RESULTS: The mean follow-up period was 45.9 months. Antibodies to HCV (anti-HCV) and HCV-RNA were tested for in each seronegative spouse every year. Seroconversion of anti-HCV occurred in only one spouse, 2 years after enrollment, with a concomitant acute hepatitis. This subject and his spouse were infected with HCV genotype 1b. Nucleotide sequence comparison of the hypervariable region of their HCV genomes showed a homology of 98%. Further phylogenetic analysis suggested that they had virtually the same isolate. Accordingly, the annual risk of interspousal transmission of HCV infection was 0.23% per year. CONCLUSIONS: These findings suggest a low incidence of interspousal transmission of HCV; however, the risk may be cumulative and such couples should be educated to avoid HCV infection from their spouses. PMID- 10824884 TI - Vaccination against hepatitis B: current challenges for Asian countries and future directions. AB - AIMS: To review the current status of hepatitis B immunization programmes as well as future issues concerning hepatitis B immunization in Asian countries. METHODS: Pertinent literature was identified via in-house and MEDLINE (1980-99) searches and references cited in published articles. Articles within the Proceedings of the IX Triennial International Symposium on Viral Hepatitis and Liver Disease provided valuable state-of-the-art resource data. RESULTS: Chronic hepatitis B infection is responsible for 75-90% of primary hepatocellular carcinoma, one of the 10 most common cancers worldwide. Hepatitis B and its chronic sequelae can potentially be eradicated through vaccines that have been shown to be 95-99% efficacious in preventing development of the disease or the carrier state in immunized infants. Approximately 75% of the world's hepatitis B carriers live in Asian countries wherein wide variations in immunization strategies exist. Vaccination programmes in hyperendemic Asian countries have elicited decreases in the incidence of acute and chronic infections as well as a decrease in chronic carriers in the unvaccinated population. Decreases in the incidence of hepatocellular carcinoma have been recorded in Taiwan and Singapore after at least 10 years of universal hepatitis B immunization programmes. CONCLUSIONS: In Asian countries currently without nationwide hepatitis B programmes, utilization of the existing vaccination infrastructure for administration of other World Health Organization Expanded Programme on Immunization vaccines will provide the most economical and efficient means of administration of the hepatitis B vaccine. PMID- 10824885 TI - Interaction between hyaluronan and CD44 in the development of dimethylnitrosamine induced liver cirrhosis. AB - BACKGROUND: A significant increase in serum hyaluronan (HA) levels has been reported in patients with liver cirrhosis. This mechanism is not yet clear, and receptors for HA have not been characterized. In this study, we examined the expression of both HA and its receptors, CD44 and intercellular adhesion molecule 1 (ICAM-1), in dimethylnitrosamine-induced liver cirrhosis. METHODS AND RESULTS: Using biotinylated HA binding protein, HA was detected in the area of periportal fibrosis and around the sinusoidal wall where hepatic fibrosis was developing. Electron microscopy revealed that HA was localized on Ito cells and sinusoidal endothelial cells (SEC). Conversely, CD44, which was only expressed weakly in normal liver, was present in large amounts in cirrhotic liver. The distribution pattern of CD44 was similar to that of HA, however, CD44 was mainly localized on the infiltrating lymphocytes and Kupffer cells. Moreover, CD44 was detected on part of factor VIII-positive SEC. Intercellular adhesion molecule-1, another receptor for HA, was detected on the surface of hepatocytes and around the sinusoidal wall in cirrhotic liver, but its distribution was not accompanied by expression of HA. With respect to CD44 isoforms, the standard form m-RNA predominated in both normal and cirrhotic liver. Variant pMeta-1 mRNA was detected at low levels. CONCLUSIONS: An interaction between HA and CD44 may play a role in the recruitment of numerous infiltrating cells and HA accumulation in hepatic sinusoids. Together with phenotypic changes in the SEC, these results may lead to a disturbance in the elimination of HA during the progression of liver cirrhosis. PMID- 10824886 TI - Clinical and haemodynamic aspects of hepatopulmonary syndrome in Indian patients with cirrhosis. AB - BACKGROUND: Hepatopulmonary syndrome consists of the triad of hepatic dysfunction and/or portal hypertension, intrapulmonary vascular dilatation and hypoxaemia, in the absence of detectable primary cardiopulmonary diseases. In the present study, we examined the frequency of hepatopulmonary syndrome among Indian patients with cirrhosis, and studied clinical predictors and pulmonary haemodynamic alterations. METHODS: Forty-five patients with cirrhosis and no cardiopulmonary diseases were investigated by air-contrast echocardiography. Where patients were positive, arterial blood gas analysis was carried out. Positive contrast echocardiography with PO2 < 70 mmHg confirmed the diagnosis of hepatopulmonary syndrome. Three cases with the syndrome and 24 without were assessed for haemodynamic status by hepatic and pulmonary catheterization. RESULTS: Four of 45 cases of cirrhosis (8.9%) had positive contrast echocardiographies, including three (6.7%) with hepatopulmonary syndrome and one 'subclinical' case (positive contrast echocardiography without hypoxaemia). Under haemodynamic study, the mean pulmonary arterial and pulmonary capillary wedge pressures appeared to be lower among those patients with hepatopulmonary syndrome. CONCLUSIONS: In this small study, the frequency of hepatopulmonary syndrome was relatively low (6.7%). Cyanosis was the only reliable clinical indicator, and there was no clear relationship with the severity of cirrhosis by Child's grading. PMID- 10824887 TI - Role of serum C-reactive protein as a marker of hepatocellular carcinoma in patients with cirrhosis. AB - BACKGROUND: The usefulness of C-reactive protein (CRP) as a tumour marker in patients with hepatocellular carcinoma (HCC) is controversial. The purpose of this study was to determine whether CRP estimation could be used to identify patients with HCC among those with cirrhosis. METHODS: Serum levels of CRP and alpha-fetoprotein (AFP) were investigated in 122 previously untreated patients with cirrhosis and HCC. Another 76 patients with cirrhosis alone were also investigated as controls. RESULTS: Of the subjects tested, 47.5% of patients with HCC and 39.5% of controls had elevated CRP values (> 6 microg/mL). Although using elevated CRP and/or AFP (> 20 ng/mL) as a criterion showed a significant difference between controls and patients with multiple nodular, massive, or diffuse type HCC (all P < 0.005), the clinical application of this criterion was limited because of low specificity (58%) and accuracy (all < 73%). By using receiver-operating characteristic curves no valuable threshold value of CRP was found to discriminate various types of HCC, except for distinguishing the diffuse type from controls. The CRP value of 12 microg/mL could be used as the cut-off value to differentiate diffuse-type HCC from controls (sensitivity 82.4%, specificity 82%, accuracy 82.1%, P<0.005). CONCLUSIONS: Serum CRP is not a good marker for HCC. However, very high values of CRP in patients with cirrhosis may suggest the presence of a diffuse-type HCC. PMID- 10824888 TI - Clinical significance of urokinase-type plasminogen activator activity in hepatocellular carcinoma. AB - BACKGROUND AND METHODS: The plasminogen activation system plays a crucial role in the process of cancer invasion and metastasis. To evaluate the most effective factor in the invasion, metastasis and prognosis of hepatocellular carcinoma (HCC), we examined urokinase-type plasminogen activator (uPA), plasminogen activator inhibitor (PAI)-1, PAI-2 and uPA activity by enzyme-linked immunosorbent assays (ELISA) in HCC tissues obtained from 46 patients. The results were compared with the patients' clinicopathological features and prognoses. RESULTS: Of the clinicopathological features, only histological portal involvement or intrahepatic metastasis, or both (INV), was significantly correlated to the disease-free survival rates (DFS; P < 0.05). The levels of uPA, PAI-1 and PAI-2 antigens were significantly associated with INV and histological grade. The DFS was not different, however, between cases with uPA, PAI-1 and PAI 2 values above and below the median. The high levels of uPA activity were closely related to INV (P < 0.001), and the activity gradually raised histological grades (P < 0.0001). The DFS was significantly different between patients with uPA activity below and above the median (0.70 ng/mL; P = 0.0092); it was also significantly different between such patients without INV (P < 0.05). CONCLUSIONS: Urokinase-type plasminogen activator activity may be the most sensitive factor affecting HCC invasion in the plasminogen activation system and a strong predictor for the recurrence of HCC. We suggest that cases with uPA activity of more than 0.70 ng/mL should be carefully followed up for possible HCC recurrence. PMID- 10824889 TI - Decreased function of peripheral blood dendritic cells in patients with hepatocellular carcinoma with hepatitis B and C virus infection. AB - BACKGROUND: Tumour immunity does not seem to be induced effectively in tumour bearing hosts, including in patients with hepatocellular carcinoma (HCC). One possible reason is that function of dendritic cells (DC) is decreased in such hosts. METHODS: We evaluated T cell stimulatory activity and interleukin (IL)-12 production of DC and interferon (IFN)-gamma and IL-10 production of T cells of peripheral blood from 12 control individuals and 21 patients with chronic hepatitis C virus (HCV) infection (six with chronic hepatitis (CH), eight with liver cirrhosis (LC) and 13 with HCC). Five hepatitis B virus (HBV)-infected patients with HCC were included as a disease control group. The DC were prepared by the culture of T cell-depleted populations of peripheral blood mononuclear cells in the presence of granulocyte-macrophage colony stimulating factor and IL 4 for a total of 11-12 days. The cytokine levels were assayed by ELISA. To test the stimulatory function of DC in T cell proliferation, mytomycin C-treated DC were cultured with allogeneic T cells from a control. RESULTS: When the T cell stimulatory activity of DC was expressed as stimulation index value of [3H] thymidine incorporation of T cells, the values were lower in HCV-infected HCC (2.6 +/- 1.8, P < 0.01) than in controls (5.5 +/- 2.0) and CH (5.0 +/- 1.3). Staphylococcus aureus Cowan 1-induced IL-12 production of DC was decreased in HCV infected HCC (P < 0.001, P < 0.01 and P < 0.05, respectively) compared with controls, CH and LC, while similar amounts of IL-10 were produced in patients and controls. Interleukin-10 and IFN-gamma production of T cells in response to anti CD3 antibody or IL-12 were equivalent between patient groups and controls, respectively. Similarly decreased DC function and normal T cell response were observed in HBV-infected HCC patients. CONCLUSIONS: These findings suggest that the depressed function of DC is associated with pathogenesis of HCC with HBV or HCV infection. PMID- 10824890 TI - Anti-neutrophil cytoplasmic antibodies in patients with chronic liver diseases: prevalence, antigen specificity and predictive value for diagnosis of autoimmune liver disease. Swedish Internal Medicine Liver Club (SILK) AB - BACKGROUND: Anti-neutrophil cytoplasmic antibodies (ANCA) against proteinase 3 are diagnostic of Wegener's granulomatosis, but ANCA occur also in patients with other inflammatory disorders, such as ulcerative colitis, primary sclerosing cholangitis (PSC) and autoimmune hepatitis. As their predictive value for autoimmune liver disease remains unknown, we analysed the prevalence and antigen specificity of ANCA in patients with various chronic liver diseases (CLD). METHODS: We studied sera from 100 patients with primary biliary cirrhosis (PBC), from 76 with PSC and from 279 with various CLD, consecutively drawn during a 5 year period at the time of liver biopsy. The ANCA were detected by indirect immunofluorescence (IIF) while the antigen specificity was characterized by ELISA by using lactoferrin, neutrophil elastase, cathepsin G and BPI (bactericidal/permeability increasing protein) as antigens. RESULTS: In PBC, ANCA were detected by IIF in 39 patients (39%). The antigen reactivity by ELISA was lactoferrin in seven, elastase in 15, BPI in 20 and cathepsin G in four patients. Four patients had reactivity against more than one antigen. In PSC, IIF demonstrated ANCA in 49 patients (65%). The antigen reactivity was lactoferrin in 17, elastase in 14, BPI in 20 and cathepsin G in four patients. Twelve patients showed reactivity against more than one antigen. In CLD, ANCA were observed in sera from 55 patients (20%). Nineteen of 45 patients (42%) with autoimmune liver disease were ANCA positive versus 36/234 (15%) with non-autoimmune liver disease (P = 0.0002). Among IIF-positive patients, antibody reactivity against lactoferrin was noted in 14, elastase in 28, BPI in 25 and cathepsin G in five patients. Twenty-one patients had reactivity against more than one antigen. Elastase and BPI antibodies occurred more frequently in patients with autoimmune compared to non-autoimmune liver disease (P < 0.01). CONCLUSIONS: Anti-neutrophil cytoplasmic antibodies are prevalent in patients with chronic liver diseases, but although they occur more frequently in patients with autoimmune liver disease their specificity and sensitivity for autoimmune liver disease is low. The predominant antigens are lactoferrin, elastase and BPI, but the correlation between IIF findings and ELISA reactivity against these antigens is weak. PMID- 10824891 TI - Cytokine profile of liver-infiltrating CD4+ T cells separated from murine primary biliary cirrhosis-like hepatic lesions induced by graft-versus-host reaction. AB - BACKGROUND AND METHODS: We have previously reported that CD4+ T cells induced primary biliary cirrhosis (PBC)-like hepatic lesions in mice with graft-versus host reaction due to major histocompatibility complex class II disparity. To clarify the relationship between the cytokine profile produced by CD4+ T cells and the formation of hepatic lesions, we sorted CD4+ T cells from the liver by using flow cytometry and examined their cytokine mRNA expression at various times after GVHR induction. We also examined the associated changes in the serum levels of antimitochondrial antibodies (AMA). RESULTS: Histologically, the infiltration of CD4+ T cells around the bile ducts was observed from day 5, and the lesions deteriorated gradually until day 14. On day 14, CD8+, B220+ and Mac-1+ cells, as well as CD4+ T cells were seen around the bile ducts. In the liver-infiltrating CD4+ T cells, the expression level of interferon-gamma (IFN-gamma) mRNA was observed to increase at an early phase (day 3), whereas that of interleukin (IL) 10 mRNA was elevated at a later phase (day 14). The elevation of IFN-gamma mRNA expression at an early phase before the appearance of non-suppurative destructive cholangitis suggests that IFN-gamma may be related to the pathogenesis of PBC in this model. Serum levels of AMA on day 14 were significantly higher than those on day 5. Interleukin-10 was considered to stimulate antibody production, to show an inhibitory effect upon the function of T helper 1 cells, and to inhibit fibrosis. CONCLUSIONS: Interferon-gamma may play an important role in the pathogenesis of this model. Moreover, delayed expression of IL-10 mRNA may control PBC-like hepatic lesions. PMID- 10824892 TI - Images of interest. Gastrointestinal: foreign bodies in the stomach. PMID- 10824893 TI - Images of interest. Hepatobiliary and pancreatic: man with recurrent jaundice and abdominal pain. PMID- 10824894 TI - Pancreatic vasoactive intestinal polypeptide-oma as a cause of secretory diarrhoea. AB - A 42-year-old woman presented with a 4-year history of worsening diarrhoea that was watery, profuse and confirmed to be secretory in nature. She had tested positive for phenolphthalein on urinary laxative screening but continued to deny laxative usage. Her vasoactive intestinal polypeptide (VIP) level was subsequently found to be markedly elevated. Despite a normal abdominal ultrasound, a computed tomography scan revealed a 5-cm pancreatic tail mass. Octreotide scanning was used to exclude metastatic disease and she went on to have surgical removal of a localized pancreatic vasoactive intestinal polypeptide oma which resulted in the complete resolution of her diarrhoea. PMID- 10824895 TI - Autoimmune pancreatitis detected as a mass in the tail of the pancreas. AB - A mass in the tail of the pancreas was detected in a 62-year-old male patient who had hypergammaglobulinaemia, and was positive for antinuclear antigen and anti-SS A antibody. Endoscopic retrograde pancreatography revealed focal irregular narrowing of the main pancreatic duct in the tail of the pancreas. Dynamic computed tomography showed swelling of the pancreatic tail, which was enhanced on delayed phase. Autoimmune pancreatitis was suspected and corticosteroid therapy was commenced. This led to significant resolution of the pancreatic stricture. It is important to recognize this clinical entity as corticosteroid therapy may avoid unnecessary surgery. PMID- 10824896 TI - Congenital heart malformations: prevalence, severity, survival, and quality of life. PMID- 10824897 TI - Evolving management for critical pulmonary stenosis in neonates and young infants. AB - Over the years, management of critical pulmonary stenosis in young infants has evolved from surgical reconstruction of the right ventricular outflow tract and closed pulmonary valvotomy to transcatheter balloon valvoplasty. Our study aimed at evaluating how the changing policy for management had affected the immediate and long term outcomes of babies with this cardiac lesion. Interventions were made in 34 infants at a median age of 8.5 days (2-90 days). Reconstruction of the right ventricular outflow tract reconstruction was performed in 10 patients, closed pulmonary valvotomy in 13, and balloon valvoplasty in 11. Initial procedure-related mortality was 50%, 15% and 0% respectively. Multivariate analysis revealed transannular patching of the right ventricular outflow tract, and male sex, to be significant factors for death. For the 27 survivors, the ratio of right ventricular to systemic systolic pressure decreased from 1.6 +/- 0.3 to 0.3 +/- 0.2 after reconstruction of the outflow tract, 1.8 +/- 0.5 to 0.8 +/- 0.4 after closed valvotomy, and 1.8 +/- 0.6 to 0.9 +/- 0.3 after balloon valvoplasty. The decrease was significantly greater after patch reconstruction (p=0.025) that required no further reinterventions. The overall rate of reintervention for the survivors was 37% (10/27). The freedom from reintervention after closed valvotomy was 82%, 64% and 51% at 1, 5 and 10 years respectively. The figure remained at 78% at both 1 and 5 years (p=0.66) after balloon valvoplasty. The higher reintervention rate for closed valvotomy corresponded to the significantly greater residual gradient across the pulmonary valve noted on follow-up (p=0.01). Reinterventions included balloon dilation (n=6), reconstruction of the outflow tract (n=4), and 1 each of ligation of an arterial duct and systemic-pulmonary arterial shunting. The risk factor for reintervention was a hypoplastic right ventricle. In conclusion, transcatheter balloon valvoplasty appears to be the optimum initial approach in view of its low mortality, efficacy at relieving the obstruction, and low rate of reintervention. PMID- 10824898 TI - Growth of children with a functionally single ventricle following palliation at moderately increased altitude. AB - The bidirectional Glenn and Fontan procedures are empirically performed as interim and definitive procedures in children with a functionally single ventricle. The optimal stage of palliation, nonetheless, remains unknown. During childhood, growth is a fundamental measure of response to therapy. Growth may be influenced by the degree of cyanosis, the volume load on the ventricle, and cardiac performance. Thus, the weight and stature of children with a functionally single ventricle who underwent a bidirectional Glenn procedure or a Fontan procedure were studied to determine the effect of each intervention on growth. Z scores for weight and stature were retrospectively determined prior to palliation, at yearly intervals for 4 years, and from long-term measurements until 18 years of age in all patients with at least 2 years of observation following palliation. Growth was evaluated in 54 patients with a bidirectional Glenn procedure, and 65 patients with a Fontan procedure. The Z scores for weight were improved after each method of surgical palliation. Stature, however, was improved only following the bidirectional Glenn procedure. Growth was impaired in patients who developed protein losing enteropathy. Weight improved only during the initial 2 years after the Fontan procedure in patients who had a surgical fenestration. Over the long-term, patients who underwent a Fontan procedure were more likely to have a Z score less than -2.0 for weight and stature than patients who underwent only a bidirectional Glenn procedure. Late mortality and the incidence of heart transplantation were increased in patients who experienced a decrease in their rate of growth, defined as a negative change of more than one Z score in weight or stature, following the Fontan procedure. In conclusion, at moderately increased altitude, children with a functionally single ventricle grow more appropriately following the bidirectional Glenn procedure than following the Fontan procedure. A decrease in the rate of growth is associated with a poor prognosis following the Fontan procedure. PMID- 10824899 TI - Similarities and differences in the exercise performance of patients after a modified Fontan procedure compared to patients with complete transposition following a Senning operation. AB - OBJECTIVE: To investigate cardiopulmonary performance in patients after a Fontan procedure, comparing it to patients following a Senning operation. METHODS: We studied 21 children, with a mean age of 11.1 years, after a total cavopulmonary anastomosis, comparing them to 13 with complete transposition after a Senning procedure, having a mean age of 11.8 years, and 21 control subjects with a mean age of 11.2 years. All were tested on a bicycle ergospirometer. RESULTS: Peak consumption of oxygen, maximal work rate, peak oxygen pulse and endexpiratory pressure of carbon dioxide at a work rate of 1.5 Watt/kg were lowest in patients with a modified Fontan procedure, and highest in the control group (p < or = 0.0278). Production of carbon dioxide, and minute ventilation at a work rate of 1.5 Watt/kg, was highest in the patients after Fontan procedure, and lowest in the healthy subjects (p < or = 0.0163). Production of carbon dioxide per single breath was lower in those having a Fontan procedure (28.9 ml) than in the two other groups (35.1 ml; p = 0.0243). The tidal volume showed no significant differences between the three groups. CONCLUSIONS: The reaction to exercise was identical qualitatively in both groups of patients, and comparable to the behaviour of patients with chronic heart failure. Quantitatively, the results of the patients following a Senning procedure lay between those of control subjects and those who had undergone a Fontan operation. The only exception was dead space ventilation, where the patients after a Fontan procedure differed from the two other groups because of their increased ventilation-perfusion mismatch. PMID- 10824900 TI - Immediate and early results of closure of moderate to large patent arterial ducts using the new Amplatzer device. AB - OBJECTIVES: Our aim was to assess the immediate and short term results of closure of moderate and large patent arterial ducts using the self-expanding and repositionable Amplatzer device. METHOD: We attempted closure in 25 patients (10 Females and 15 males) using the Amplatzer occluder. Their median age was 48 months with a range from 8 months to 26 years and median weight of 14 kg with a range from 4.5 kg to 48 kg. The mean ductal diameter was 4.1 mm (S.D 1.51 mm). A 6F/7F long sheath was used to deliver the device. Follow up was performed with colour- flow mapping of the pulmonary trunk within 24 hours, at 3 months, and 6 months of closure. RESULTS: Of the 25 patients, the device was placed successfully in 23. Concurrent angiography showed immediate closure in 12 patients, while 8 had trivial shunting and 3 had mild shunting. Within 24 hours, Doppler examination revealed complete closure in all but three patients, who had a mild residual shunt. Two attempts were unsuccessful. Both these patients underwent successful surgical ligation. All except one patient were discharged on the next day. Of the 23 patients, 15 (65%) have been followed up for 3 months, while 8 (35%) have completed 6 months of follow-up. Of the three patients initially with mild residual flow, two had completely closed at 3 months The one remaining patient is yet to be seen at the 3 month follow-up. Thus, at 3 months, all patients studies had shown complete closure. CONCLUSION: Antegrade transcatheter closure using the Amplatzer duct occluder is an efficacious treatment for bigger patent arterial ducts. Long-term follow-up is necessary to show sustained benefits and confirm the absence of side effects. PMID- 10824901 TI - Surgical approach to complicated cervical aortic arch: anatomic, developmental, and surgical considerations. AB - BACKGROUND: Abnormalities of brachiocephalic arterial branching and arch laterality are common in patients with a cervical aortic arch. In addition, structural anomalies of the arch such as obstruction, aneurysms, and tortuosity are found in a significant number of cases. METHODS: Between 1990 and 1998, 6 patients underwent surgery for an obstructed right cervical arch. A significant obstruction was present at the transverse or distal arch in all patients, and was recurrent after previous repair in 2. In 1 patient, there was also a multi-lobed aneurysm of the aortic segment contiguous to the obstruction, and in 2 there was marked tortuosity of the arch. In all cases, the order of origin of the head and neck vessels was abnormal, and obstruction of 1 or more brachiocephalic vessels was found in 3. A vascular ring was present in all patients, with a right aortic arch and aberrant left subclavian artery in 4 patients and a double aortic arch with a dominant right cervical arch in 2. The descending aorta was circumflex (left-sided) in 3 patients. Three patients were repaired through a standard right posterolateral thoracotomy, and 3 through a median sternotomy. Patch augmentation aortoplasty was used in 2 patients, a tube graft from the ascending to descending aorta in 2, end to side anastomosis of the descending aorta to the proximal arch in 1, and direct anastomosis to reconstruct an atretic left-sided component of a double arch in 1. RESULTS: Repair was successful in all cases, with no perioperative complications. At follow-up ranging from 1 to 9 years, all patients were alive and well, with no recurrence of arch obstruction or other significant complications. Fluorescent in situ hybridization revealed microdeletion of chromosome 22q11 in 1 patient (not performed in the others). CONCLUSIONS: Structural anomalies of the arch are relatively common in patients with a cervical aortic arch. Such abnormalities may be the result of hemodynamic conditions and/or abnormal vascular tissue related either to the cervical position of the arch or its embryologic precursors. Given the highly variable anatomy of patients with a complicated cervical aortic arch, surgical considerations will vary in kind. PMID- 10824902 TI - Diagnosis of left juxtaposition of the atrial appendages in the fetus. AB - We describe two cases of left juxtaposition of the atrial appendages in which fetal echocardiograms provided a clue to the diagnosis. Both cases were associated with complex cyanotic congenital heart disease. The clue to the diagnosis was found at the three-vessel view. Abnormal vascular spaces were seen on the left side of the cross-sections of the great arterial trunks. PMID- 10824903 TI - Balloon aortic valvotomy through a carotid cutdown in infants with severe aortic stenosis: results of the multi-centric registry. AB - OBJECTIVES: The purpose of this study was to evaluate the short and intermediate term results of infants who have undergone balloon aortic valvotomy from the carotid arterial approach, and to identify risk factors in those infants who had a poor outcome. METHODS: Between 1988 and 1999, balloon aortic valvotomy was attempted at four centres in 95 infants with severe aortic stenosis. Echocardiographic and hemodynamic data, and outcome, were analysed retrospectively. RESULTS: Valvotomy was accomplished in 92 of the 95 infants, with a median age of 5 days, a range from 0 to 191 days, and weighing 3.4 kg, with a range from 1.0 to 6.5 kg. Major procedural complications occurred in 10 infants. Post-procedural aortic regurgitation was severe in 5 patients. There were 13 early deaths, and 4 late deaths. The period of mean follow-up has been 2.1 years, with a range from 0 to 9.3 years. The actuarial survival at 3 years was 76 +/- 6%. Further interventions were needed in 19 patients, giving a 3-year freedom from reintervention of 67 +/- 6%. The 51 infants who were duct-dependent were further analyzed, and found to have a higher mortality (38%) compared to those infants not dependent on the arterial duct (5%). Risk factors for a poor outcome in the duct-dependent infants were mitral stenosis (p<0.005), a left ventricle which did not form the cardiac apex (p<0.005), and an aortic valve with a diameter of less than 6 mm (p<0.05). CONCLUSIONS: This multi-centric registry shows good results in the intermediate term for treating infants with severe aortic valvar stenosis with balloon valvotomy through a carotid arterial cutdown. Infants dependent on prostaglandin had a worse outcome, especially if they had any of the identified risk factors. PMID- 10824904 TI - Embryology of congenital ventriculo-coronary communications: a study on quail chick chimeras. AB - Ventriculo-coronary arterial communications are rare congenital heart defects which have been explained traditionally on the basis of abnormal persistence of such communications found in the normal developing heart. Recent studies, however, have suggested that these embryonic communications might be an incidental finding rather than a normal feature. Thus, it has been suggested that congenital ventriculo-coronary communications do not represent remnants of normal embryonic vessels, but rather represent acquired lesions. In the present study, hearts were constructed in embryonic chicks in which the coronary vasculature was almost completely derived from a quail-donor. After immunohistochemical staining of the quail-derived coronary endothelium, chimeric hearts were analysed with respect to the presence of embryonic ventriculo-coronary communications, and with respect to the origin of these structures from either coronary arteries or endocardium. The results demonstrate the normal presence of ventriculo-coronary communications in avian embryonic hearts. They show, furthermore, that these structures are of coronary endothelial origin. The findings are in accord with the traditional view on the pathogenesis of congenital ventriculo-coronary communications. The roles of elevated ventricular pressure, abnormal remodelling of the developing myocardium, and of abnormal growth of the coronary vasculature are discussed relative to the pathogenesis of congenital ventriculo-coronary communications. PMID- 10824905 TI - Psychological distress and styles of coping in parents of children awaiting elective cardiac surgery. AB - AIMS: We sought to assess the level of psychological distress, and the styles of coping of, parents of children with congenital heart disease. The study was based on questionnaires, which were completed, on average, four weeks, with a range from 0.1 to 22.1 weeks, prior to elective cardiac surgery or elective catheter intervention. METHODS: We used the General Health Questionnaire, and the Utrecht Coping List, to compare scores from parents of those undergoing surgery, with scores of reference groups, and with scores of the parents of those undergoing intervention. RESULTS: Overall, in comparison with our reference groups, the parents of the 75 children undergoing surgery showed elevated levels of psychological distress, manifested as anxiety, sleeplessness, and social dysfunctioning. They also demonstrated less adequate styles of coping, being, for example, less active in solving problems. With only one exception, no differences were demonstrated in parental reactions to whether cardiac surgery or catheter intervention had been planned. The mothers of the 68 patients who were to undergo cardiac surgery, however, reported greater psychological distress and manifested greater problems with coping than did the fathers. CONCLUSION: Elevated levels of psychological distress, and less adequate styles of coping, were found in the parents of patients about to undergo cardiac surgery, especially the mothers, when compared to reference groups. Future research should investigate whether these difficulties persist, and whether this will influence the emotional development of their children with congenital cardiac malformations. PMID- 10824906 TI - Images in congenital heart disease. Accessory atrioventricular valvar tissue in congenitally corrected transposition. PMID- 10824907 TI - Can we describe structure as well as function when accounting for the arrangement of the ventricular mass? PMID- 10824908 TI - Is there a possible role for haemostasis in the development of perigraft reaction complicating the modified Blalock Taussig shunt? AB - The perigraft reaction is an unusual complication found in patients in whom a modified Blalock Taussig shunt has been created using a polytetrafluoroethylene graft. We found that, in two infants, consistent laboratory findings during such a perigraft reaction were hypofibrinogenemia, increased levels of thrombin antithrombin III complex, prothrombin fragment 1 and 2 and products of degradation of fibrin. Normalization of the levels of fibrinogen produced resolution of the perigraft reaction. PMID- 10824909 TI - Severe haemolysis after transcatheter closure of a patent arterial duct with the new Amplatzer duct occluder. AB - Severe mechanical haemolysis occurred in an 11-month-old boy after implantation of the new Amplatzer duct occluder. Temporary balloon occlusion of the aortic ampulla was performed 4 days after the initial procedure leading to prompt abolition of the haemolysis. PMID- 10824910 TI - Congenitally corrected transposition with pulmonary atresia and intact ventricular septum. AB - We describe a patient with the rare association of the heart in the left chest, congenitally corrected transposition, pulmonary atresia and an intact ventricular septum. There were associated fistulous communications between the morphologically left ventricle and the coronary arteries. Diagnosis was made by echocardiography, and subsequently confirmed by cardiac catheterization. PMID- 10824911 TI - Rapid recurrence of an inflammatory myofibroblastic tumor in the right ventricular outflow tract. AB - We describe an unusual case of rapid recurrence of a previously excised inflammatory myofibroblastic tumor of the right ventricular outflow tract in a 5 month-old infant. The infant is asymptomatic 18 months after the second surgery. The very rare cardiac involvement, and the early relapse of the inflammatory pseudotumor, to the best of our knowledge, is a unique combination. The inflammatory myofibroblastic tumor, as known as a pseudotumor or plasma cell granuloma is an uncommon reactive lesion with unknown aetiology. It is found most commonly in the lung and a number of visceral organs, such as the spleen, liver, ileum, salivary glands, urinary bladder, larynx or brain or in the retroperitoneum and lymph nodes. To our knowledge only 9 cases have been published of such tumor arising within the heart. PMID- 10824912 TI - Pulmonary arterial thrombosis in a neonate with homozygous deficiency of antithrombin III: successful outcome following pulmonary thrombectomy and infusions of antithrombin III concentrate. AB - We report a newborn male who presented with severe central cyanosis on the third day of life. Partial thrombotic obstruction of the pulmonary trunk secondary to Antithrombin III (homozygous defect of heparin binding site) deficiency was subsequently diagnosed. Surgical thrombectomy, and infusions of Antithrombin III concentrate, led to a successful outcome. We postulate that intrauterine thrombosis occurred to give this unusual presentation. PMID- 10824913 TI - Banding of a patent arterial duct to palliate complete transposition in a compromised neonate. AB - In a hemodynamically compromised neonate with complete transposition and intact ventricular septum (concordant atrioventricular and discordant ventriculo arterial connections), it became necessary to control severe congestive heart failure. The arterial switch operation had been delayed because of intracranial bleeding and subsequent coagulopathy. The cardiac failure was reversed by surgical banding of the patent arterial duct. The arterial switch was performed successfully one week later. PMID- 10824914 TI - Vascular aneurysm producing divided right atrium in a patient with pulmonary atresia and intact ventricular septum. AB - We describe a patient with pulmonary atresia and intact ventricular septum in whom the right atrium was divided by a vascular aneurysm located in the right atrioventricular groove. We postulate that the structure represents an aneursymally dilated right coronary artery taking anomalous origin from the pulmonary trunk, with fistulous communication to the right atrium. We discuss the findings relative to concepts of development of the coronary arteries in normal hearts and in pulmonary atresia with an intact ventricular septum. PMID- 10824915 TI - An update on the state of paediatric cardiology and cardiac surgery in Europe. PMID- 10824916 TI - Prevalence of small muscular defects is dependent upon the way in which the babies are examined. PMID- 10824917 TI - Prevalence and prognostic significance of neuroendocrine cells in esophageal adenocarcinoma. AB - Neuroendocrine differentiation is common in adenocarcinomas of the stomach and colon and may be associated with a slightly better prognosis in gastric adenocarcinoma. We studied neuroendocrine differentiation in esophageal adenocarcinomas and associated Barrett's esophagus (BE) to determine association with patient outcome. Fifty-eight cases of esophageal adenocarcinoma (15 biopsies, 43 resections) from 52 patients were stained with a monoclonal antibody to chromogranin (CG). Medical records were reviewed for tumor stage, response to therapy, and patient survival. Thirty-two patients received radiation and chemotherapy, and four received radiation. Twelve of 58 (20.7%) esophageal adenocarcinomas contained scattered CG-positive cells. Tumors with CG-positive cells were moderately to poorly differentiated, and many consisted of large cribriform glands, similar to intestinal-type adenocarcinomas. One case of small cell carcinoma of the esophagus was weakly CG positive; another was negative. Neuroendocrine differentiation was retained in lymph node metastases in two cases but lost in three other cases. In 10 CG-negative primary tumors, lymph node metastases were also negative. For five of six patients with paired biopsy/resection specimens, no CG-positive cells were seen in either specimen; one patient had CG-positive cells only in the resection. There was no difference in tumor stage at surgery or survival time between CG-positive and CG-negative tumors. BE was present in 34 cases and contained CG-positive cells in 21 of 34 (61.8%). Low-grade dysplasia contained CG-positive cells in 11 of 14 cases (78.6%) and high-grade dysplasia in 3 of 6 cases. Fourteen of 21 (66.7%) adenocarcinomas associated with CG-positive BE were negative for CG. In summary, neuroendocrine differentiation is common in BE and is retained in low- and high grade dysplasia but is usually lost in esophageal adenocarcinoma. The presence of scattered neuroendocrine cells does not affect patient outcome. PMID- 10824918 TI - Association of SYT-SSX fusion types with proliferative activity and prognosis in synovial sarcoma. AB - The t(X;18)(p11.2;q11.2) translocation commonly found in synovial sarcoma (SS) results in the fusion of the SYT gene on chromosome 18 to either of two closely related genes, SSX1 and SSX2, on chromosome X. It has been suggested that patients who have SS bearing SYT-SSX1 fusion have worse prognosis than those bearing SYT-SSX2 fusion. However, little is known about the biologic basis or the relationship with the histopathologic risk factors in regard to the different fusion types. We analyzed 19 cases of SS with no metastasis at diagnosis. These tumors were classified by reverse transcription-polymerase chain reaction to SYT SSX1 and SYT-SSX2 types. The expression of Ki-67, p27, p53, and bcl-2 and various clinicopathologic parameters including mitotic rate were compared between the two fusion types. The SYT-SSX1 type fusion was associated with high Ki-67 expression (P = .011) and high mitotic rate (P = .070). No significant differences were found between the two types as to the expression of p27, p53, and bcl-2 and other clinicopathologic parameters. The survival analysis showed that SYT-SSX1-type fusion, high Ki-67 expression, and high mitotic rate correlated with shorter metastasis-free survival. These data suggested that SYT-SSX fusion type is associated with tumor cell proliferative activity and prognosis of patients who have SS. PMID- 10824919 TI - Primary neuroendocrine carcinoma (thymic carcinoid) of the thymus with prominent oncocytic features: a clinicopathologic study of 22 cases. AB - Twenty-two cases of oncocytic thymic neuroendocrine carcinomas (carcinoid tumors) are presented. The patients were 17 men and 5 women between the ages of 26 and 84 years (median, 55 years). Nine were asymptomatic, and the tumor was found on routine examination; four patients presented with chest pain, two with weight loss, two with multiple endocrine neoplasia I syndrome, and one with Cushing's syndrome. Surgical resection of the mediastinal tumor was performed in all cases. The lesions were described as soft, light tan to brown, measuring from 3 to 20 cm in greatest diameter. On cut section, the tumors showed a homogeneous surface, soft consistency, and focal areas of hemorrhage. Microscopically, the lesions were characterized by nests or trabeculae of tumor cells that contained abundant granular to densely eosinophilic cytoplasm, with round to oval nuclei and in some areas prominent nucleoli. Mitotic figures ranged from 2 to 10 per 10 high-power fields; foci of comedonecrosis were seen in all cases. Immunohistochemical studies including broad spectrum keratin, CAM 5.2, chromogranin, synaptophysin, Leu-7, and p53 were performed in 12 cases. All of the tumors were strongly positive for CAM 5.2 low-molecular-weight cytokeratin, 11 showed strong positive reaction for Leu-7, 10 for broad-spectrum keratin, 8 for chromogranin, 7 for synaptophysin, and only 1 case showed focal positive staining of the tumor cells for p53. Clinical follow-up of 14 patients showed that 10 were alive between 2 and 11 years, and 4 patients had died of tumor from 4 to 11 years after diagnosis. Patients with good clinical outcome were those whose tumors showed low mitotic activity and minimal nuclear pleomorphism, whereas those who had died of their tumors were those whose tumors were characterized by marked nuclear atypia and higher mitotic rates. Oncocytic thymic carcinoids should be added to the differential diagnosis of anterior mediastinal neoplasms characterized by a monotonous population of tumor cells with prominent oncocytic features. PMID- 10824920 TI - The coexistence of low-grade mucinous neoplasms of the appendix and appendiceal diverticula: a possible role in the pathogenesis of pseudomyxoma peritonei. AB - We examined 38 appendectomies with diagnoses of mucocele, diverticulum, or adenoma to study the coincidence of appendiceal diverticula and appendiceal low grade mucinous neoplasms and to examine the possible role of diverticula in the pathogenesis of pseudomyxoma peritonei. Invasive adenocarcinomas and retention cysts were excluded (six cases). Cases were classified as adenomas or mucinous tumors of unknown malignant potential, with or without diverticula. Medical records were reviewed for multiple parameters, including presenting symptoms, presence of pseudomyxoma peritonei, and presence of associated malignancies. Binomial statistics were used to calculate the probability that the observed prevalence of low-grade mucinous neoplasms and diverticula together was significantly different from the expected prevalence of diverticula or low-grade mucinous neoplasms alone, using historical controls from the literature. Twenty five percent of the total cases (8 of 32) contained both a low-grade mucinous neoplasm (7 cystadenomas and 1 mucinous tumor of unknown malignant potential) and a diverticulum. Thus, 8 of 19 low-grade mucinous neoplasms (42%) were associated with diverticula. Of the appendices with both low-grade mucinous neoplasms and diverticula, three contained dissecting acellular mucin within the appendiceal wall, four showed diverticular perforation, and one had pseudomyxoma peritonei associated with the ruptured diverticulum. A significant percentage (P < .001) of cases contained low-grade mucinous neoplasms and diverticula together. The case of coexistent low-grade mucinous neoplasm, diverticulum, and pseudomyxoma peritonei suggests that diverticula could play a role in the pathogenesis of pseudomyxoma peritonei. This could occur either by involvement of preexisting diverticula by the neoplasm or by distention of the appendiceal lumen by mucin, leading to increased intraluminal pressure and subsequent diverticulum formation at a weak area in the wall. Either mechanism might allow low-grade mucinous neoplasms to penetrate the appendiceal wall more easily. PMID- 10824921 TI - Pagetoid reticulosis (Woringer-Kolopp disease): an immunophenotypic, molecular, and clinicopathologic study. AB - Pagetoid reticulosis (PR), also known as Woringer-Kolopp disease, is a form of cutaneous T-cell lymphoma that demonstrates striking epidermotropism on histologic examination. We present the histologic, immunologic, and molecular findings for seven patients who had PR. The patients ranged in age from 33 to 67 years. All patients presented with one or several thick plaques involving the distal extremities except for one patient, who presented with a tongue lesion. Immunohistochemical staining of the atypical lymphoid cells demonstrated a T-cell phenotype in all cases. In one of four frozen cases, the neoplastic cells were of T-helper cell phenotype (CD4 positive). Four of seven cases demonstrated a T cytotoxic/suppressor cell phenotype (CD8 positive). The T-cell subset for the remaining two cases could not be determined. CD30 positivity and a high growth fraction as indicated by staining with Ki-67 were seen in three of seven and three of four cases, respectively. Genotypic analysis performed on three of our cases revealed T-cell receptor (gamma and/or beta) rearrangement, indicating a clonal proliferation. The clinical follow-up ranged from 15 months to 13 years. Four of seven patients are alive and free of disease after treatment with excision or local irradiation. One patient relapsed twice after treatment with radiation and photochemotherapy with 8-methoxypsoralen and UVA and was then lost to follow-up. The lesions of another patient resolved spontaneously but recurred at the same and in an additional site 5 years later. One patient recurred after electron beam therapy. The recurrent lesion improved with radiation therapy and local wound care but never resolved completely. The patient died of unrelated causes. Our findings suggest that PR is a distinct clinicopathologic entity, separate from unilesional mycosis fungoides, demonstrating a slow disease course. The disease is a clonal cutaneous T-cell lymphoma with relatively consistent clinical and histopathologic findings but a heterogeneous immunophenotypic profile. PMID- 10824922 TI - Prognostic values of stromal proportion and PCNA, Ki-67, and p53 proteins in patients with resected adenocarcinoma of the lung. AB - Data from 64 patients who underwent surgical resection of lung adenocarcinomas were studied to identify clinicopathologic markers that might provide prognostic information on the clinical behavior of this neoplasia Patient staging was performed in accordance with the tumor-node-metastasis system as follows: Stage I (n = 29), Stage II (n = 11), Stage IIIA (n = 21), and Stage IIIB (n = 3). Overall follow-up time corresponded to the follow-up time for patients who were alive and to the survival time for patients who had died, all of them expressed in months. Data included age, staging, histologic type, morphometric assessment of histologic features related to tumor (stroma and vascularization), and immunohistochemical detection of proliferation cell markers (Ki-67 protein and proliferating cell nuclear antigen) and p53 protein. The morphometric assessment was made by the point-counting procedure. Data analysis included Life Tables for Survival and Cox Regression models. Overall follow-up analysis showed that significant univariate predictors (P < .05) were T stage; N stage; tumor stromal proportion; and immunohistochemical indexes of proliferating cell nuclear antigen, Ki-67, and p53 proteins. Variables that presented independent predictive value for overall follow-up with the multivariate model (P < .05) were sex, T stage, N stage, tumor stromal proportion, and immunohistochemical detection of p53 protein. We conclude that tumor stromal proportion and immunohistochemical detection of p53 protein, controlled for sex, T stage, and N stage, may be of critical value in the evaluation of recurrence of lung adenocarcinoma, serving as indicators for a more accurate prognosis. PMID- 10824923 TI - Needle biopsy of recurrent adenocarcinoma of the prostate after radical prostatectomy. AB - The objective of this study was to evaluate needle biopsy of recurrent prostate cancer after radical prostatectomy. We evaluated 37 cases of recurrent prostate cancer after radical prostatectomy that were diagnosed by needle biopsy between March 1984 and July 1998. Fifteen were from consultations in which contributors were uncertain of the diagnosis, and 22 were from men who had come to The Johns Hopkins Hospital for treatment. The median interval from radical prostatectomy to biopsy showing recurrent tumor was 40 months. There was no correlation between the interval to recurrence and either pathologic features of the biopsy and radical prostatectomy or various clinical features. The mean extent of adenocarcinoma in the biopsies was 3.2 mm (range, 0.1 to 18 mm; median, 2 mm). The length of recurrent cancer on biopsy correlated with an abnormal rectal examination (P = .001). The mean Gleason score for the recurrent tumors was 6.5, which correlated with the grade of the radical prostatectomy cancer (P = .005). The cancers often lacked overt histologic features of malignancy. Benign prostatic acini were seen in five cases (14%), usually separate from the cancer. In 5 (33%) of the consultation cases, we would not have been able to diagnose cancer if not for the fact that atypical prostate glands should not be present after radical prostatectomy. In well-sampled radical prostatectomies, margins were almost always positive, as was extraprostatic extension. In cases with less sampling, there was a higher incidence of organ-confined disease and margin negative disease implying suboptimal processing of the radical prostatectomy. After radical prostatectomy, recurrent cancer on needle biopsies may be focal and difficult to diagnose and must be assessed differently than in patients who have not had surgery. PMID- 10824924 TI - Amyloidosis of the larynx: a clinicopathologic study of 11 cases. AB - Laryngeal amyloidosis (LA) is uncommon and poorly understood, with limited long term clinicopathologic and immunophenotypic studies in the literature. Eleven cases of LA were retrieved from the files of the Otorhinolaryngic-Head & Neck Tumor Registry from 1953 to 1990. The histology, histochemistry, immunohistochemistry, and follow-up were reviewed. All patients (three women and eight men) presented with hoarseness at an average age of 37.8 years. The lesions, polypoid or granular, measured an average of 1.6 cm and involved the true vocal cords only (n = 4), false vocal cord only (n = 1), or were transglottic (n = 6). An acellular, amorphous, eosinophilic material was present in the stroma, often accentuated around vessels and seromucous glands, which reacted positively with Congo red. A sparse lymphoplasmacytic infiltrate was present in all cases that demonstrated light chain restriction by immunohistochemistry in three cases (kappa = 2, lambda = 1). Serum and urine electrophoreses were negative in all patients. Treatment was limited to surgical excision, including a single laryngectomy. Six patients manifested either recurrent and/or multifocal/systemic disease: two patients with light chain restriction were dead with recurrent disease (mean, 11.1 years); two patients were dead with no evidence of disease (mean, 31.7 years); and two patients were alive, one with light chain restriction and recurrent and multifocal disease (41.6 years) and one with no evidence of disease after a single recurrence (43.4 years). The remaining five patients were either alive or had died with no evidence of disease an average of 32.4 years after diagnosis. No patient developed multiple myeloma or an overt B-cell lymphoma. LA is an uncommon indolent lesion that may be associated with multifocal disease (local or systemic). The presence of an associated monoclonal lymphoplasmacytic infiltrate and recurrent/multifocal disease in the respiratory or gastrointestinal tract of a few cases and the lack of development of a systemic plasma cell dyscrasia or overt systemic B-cell malignancy suggest that some LA may be the result of an immunocyte dyscrasia or tumor of mucosa-associated lymphoid tissue. PMID- 10824925 TI - KIT expression in angiosarcomas and fetal endothelial cells: lack of mutations of exon 11 and exon 17 of C-kit. AB - C-kit proto-oncogene product (KIT, CD117) is a tyrosine kinase growth factor receptor for stem cell factor. This receptor is important for the development and maintenance of hematopoietic stem cells, mast cells, germ cells, melanocytes, and interstitial cells of Cajal and is constitutively expressed in them. Among mesenchymal tumors, KIT seems to be specific for the gastrointestinal stromal tumors, which consistently express this protein. Activating mutations in the tyrosine kinase or juxtamembrane domains of c-kit gene have been found in mastocytoma, seminoma, and gastrointestinal stromal tumors. Following up our initial observation of KIT expression in one angiosarcoma, we examined 50 angiosarcomas, 13 Kaposi sarcomas, 10 epithelioid hemangioendotheliomas, and 31 hemangiomas of different types for KIT expression using a polyclonal antiserum specific to KIT. Adult and fetal tissues and neovascular endothelia in 20 carcinomas were studied for comparison. More than half (56%) of the angiosarcomas representing different clinicopathologic and histologic subtypes and 2 of 13 Kaposi sarcoma were KIT positive. All epithelioid hemangioendotheliomas and hemangiomas were negative, with the exception of two infantile hemangiomas that showed KIT reactivity. The fetal capillary endothelia of lungs, placenta, and soft tissues were also KIT positive, although in soft tissues and placenta, KIT positivity was more prominent in the first trimester. However, endothelia of adult vessels and neovascular capillaries of carcinomas were negative. None of the four KIT-positive angiosarcomas and one KIT-positive Kaposi sarcomas that were studied showed mutations in the juxtamembrane or tyrosine kinase domains of the c-kit gene. These results indicate that KIT expression occurs in a subset of angiosarcomas, and the expression probably represents oncofetal expression (i.e., reversion of the tumor cell phenotype to that of fetal endothelial cells that may show KIT expression). PMID- 10824926 TI - Topoisomerase IIalpha expression in breast cancer: correlation with outcome variables. AB - Topoisomerase IIalpha (topo IIalpha) plays a key role in DNA replication and is a target for multiple chemotherapeutic agents. In breast cancer, topo II expression has been linked to cell proliferation and HER2/neu protein overexpression. However, its relationship with outcome variables is not well established. Formalin-fixed, paraffin-embedded primary breast cancers from 184 women (mean age, 60 years) were stained for topo II by automated immunohistochemistry. A topo II expression index (TI) was determined by counting the number of positive cells per high-power field and calculating an overall mean number of positive cells per high-power field. Tumors with a TI of more than 1 were considered positive, and those with a TI of 1 or less were considered negative. A cell proliferation index was determine d by automated immunohistochemistry using the MIB-1 antibody in an identical technique. HER-2/neu gene amplification (HER-2 amp) was determined by automated fluorescence in situ hybridization using the Ventana unique sequence probe. Fifty-nine (32%) of the tumors had a TI greater than 1. On univariate analysis, increased topo II expression correlated with decreased patient survival (p = .001), advanced tumor stage (p = .034), lymph node metastasis (p = .018), and HER-2 amp (p = .016). Tumor stage (p < .0001), node-positive status (p < .0001), tumor grade (p = .025), HER-2 amp (p < .0001), and MIB-1 overexpression (p = .002) also correlated with survival on univariate analysis. Topo II expression did not correlate with tumor size, grade, estrogen receptor/progesterone receptor status, or disease recurrence. On multivariate analysis, stage (p < .0001), lymph node metastasis (p < .0001), and tumor grade (p = .002) all independently predicted disease-related death. Increased topo II expression is associated with an aggressive form of breast cancer featuring HER-2 amp and predicts disease-related death, lymph node metastasis, and advanced tumor stage. PMID- 10824927 TI - Amplification and overexpression of mdm2 gene in ependymomas. AB - Ependymomas rarely show p53 gene alteration, and the tumorigenic mechanism of ependymomas still remains to be elucidated. We investigated the amplification and overexpression of mdm2 gene, whose product (MDM2) is considered to be one of the major cellular regulators of p53-mediated growth control, in 26 specimens of ependymomas obtained from 20 patients. The majority of the ependymomatous samples (96%) showed at least focal immunopositivity for MDM2; however, only 8% of the samples were immunopositive for p53. mdm2 gene amplification was detected in 35% of the samples by differential polymerase chain reaction, all of which overexpressed MDM2. These results suggest that the amplification and/or overexpression of mdm2 may be one of the major molecular events occurring in the tumorigenesis of ependymomas. PMID- 10824929 TI - Retinoblastoma expression in thyroid neoplasms. AB - Retinoblastoma (Rb) mutation in thyroid neoplasia has been identified in a few molecular studies; however, the utility of Rb immunohistochemistry in distinguishing benign and malignant thyroid lesions has not been documented in formalin-fixed, paraffin-embedded tissues. The present study investigated Rb immunohistochemistry in a series of 111 formalin-fixed, paraffin-embedded benign and malignant thyroid lesions. All of the major histologic subtypes were included to detect any heterogeneity in Rb-1 expression that might influence the diagnostic utility of this technique or further elucidate the pathogenesis of thyroid neoplasia among the categories. Altogether, 34 follicular adenomas, 9 follicular carcinomas, 7 Hurthle cell adenomas, 5 Hurthle cell carcinomas, 23 papillary carcinomas (8 of which were follicular variants), 4 insular carcinomas, 4 anaplastic carcinomas, 6 medullary carcinomas, and 19 nodular goiters were analyzed. Avidinbiotin immunohistochemistry was performed using the Dako Rb-1 clone. Pronase digestion was introduced into the epitope retrieval protocol to eliminate false-positive cytoplasmic stainig. Nuclear immunoreactivity was assessed as positive if 10% or more of thyroid epithelial nuclei stained positively, and conversely as negative. The majority of benign non-Hurthle thyroid lesions, whether hyperplastic or neoplastic, retained Rb nuclear immunopositivity in most cells (51 of 53 cases [96%]). Conversely, malignant thyroid neoplasms lacked Rb immunoreactivity in the majority (42 of 51 cases [82%]), including all papillary carcinomas (23 of 23) and almost all follicular carcinomas (8 of 9 [89%]). Virtually all Hurthle cell neoplasms were negative (11 of 12 [92%]), whether benign or malignant. In conclusion, Rb immunohistochemistry can aid in the distinction between benign and malignant thyroid lesions in conjunction with morphology. This seems to be most applicable to the often problematic differentiation between follicular adenoma and the follicular variant of papillary carcinoma (P < .0001; sensitivity and specificity, 100%) or minimally invasive follicular carcinoma (P = .0007; sensitivity, 89%; specificity, 100%). PMID- 10824928 TI - Primary large-cell neuroendocrine carcinoma of the parotid gland: immunohistochemical and molecular analysis of two cases. AB - Pulmonary large-cell neuroendocrine carcinoma (LCNEC) is a newly proposed clinicopathologic entity; a few cases of LCNEC have been reported in other sites, such as the uterine cervix and the thymus. In the salivary glands, LCNEC is extremely rare and is not recognized as a specific entity in the World Health Organization classification. We retrospectively reviewed from our files 1675 cases of surgically resected primary parotid gland tumors and found 2 cases of LCNEC that fulfilled the criteria of pulmonary LCNEC. These cases occurred in 72- and 73-year-old men who had short histories of enlarging parotid gland tumors. The tumors were composed of large cells that exhibited organoid, solid, trabecular, and rosette-like growth patterns with a high mitotic rate and a conspicuous tendency for necrosis. The tumor cells were polygonal and characterized by a moderate nuclear:cytoplasmic ratio, coarse chromatin, and conspicuous nucleoli. Immunohistochemical examination revealed that the tumor cells were positive for six general neuroendocrine markers, cytokeratin, p53, bcl 2, epidermal growth factor receptor, and cyclin D1. Markedly reduced expressions of p21Waf1 and p27Kip1 were also noticed. The Ki-67 labeling index was more than 50% in both cases. One case showed loss of heterozygosity at TP53 accompanied by a p53 gene point mutation. Loss of heterozygosity at chromosome 9p21 was detected in both cases; one was accompanied by a p16 gene silent point mutation. Both patients died of the disease, with recurrence 5 months and 4 years after surgery, respectively. These findings indicate that LCNEC is a rare but distinct salivary gland tumor with highly aggressive biologic behavior. Multiple alterations of cell cycle regulators and tumor suppressor genes may play an important role in presenting the biologic characteristics of this rare parotid gland tumor. PMID- 10824930 TI - Thyroid transcription factor-1 in normal, hyperplastic, and neoplastic follicular thyroid cells examined by immunohistochemistry and nonradioactive in situ hybridization. AB - Thyroid transcription factor-1 (TTF-1) has been known to regulate the transcriptional activity of thyroid-specific genes. We examined the expression of TTF-1 in non-neoplastic and neoplastic thyroid tissues. By immunohistochemistry, the nuclei of normal and hyperplastic follicular cells strongly reacted with the antibody against TTF-1. Immunohistochemistry also revealed a distinctive nuclear positivity of TTF-1 in all 33 differentiated follicular cell tumors, including 15 follicular adenomas, 5 follicular carcinomas, and 13 papillary carcinomas. No immunoreactions were observed in three of four undifferentiated carcinomas, whereas an isolated and weak nuclear positivity was obtained in one. In normal and hyperplastic tissues, the distribution of TTF-1 was fairly related to that of thyroid-specific proteins thyroglobulin and thyroperoxidase. However, discrepancies in the distribution were observed in tumor tissues. By in situ hybridization, the riboprobe hybridized distinctively with the cytoplasm of neoplastic cells as well as normal follicular cells. Papillary carcinoma cells expressed TTF-1 mRNA in all but two cases, and its expression was also demonstrated in one of four undifferentiated carcinomas. Reverse transcription polymerase chain reaction confirmed the presence of TTF-1 mRNA in two human undifferentiated carcinoma cell lines, TTA-1 and TTA-2. In conclusion, the investigation of TTF-1 provides useful information on the functional activities and/or differentiation of thyroid tumors and may lead to an increase in our understanding of the biologic nature of thyroid tumors. PMID- 10824931 TI - Extragastrointestinal (soft tissue) stromal tumors: an analysis of 48 cases with emphasis on histologic predictors of outcome. AB - The clinicopathologic features of 48 tumors that were histologically similar to gastrointestinal stromal tumors but occurred in the soft tissues of the abdomen were analyzed to determine their overall similarity to their gastrointestinal counterpart, their biologic behavior, and the parameters that predict risk for adverse outcome. Classic leiomyomas and leiomyosarcomas were specifically excluded. The tumors occurred in 32 women and 16 men, who ranged in age from 31 to 82 years (mean, 58 years). Forty tumors arose from the soft tissue of the abdominal cavity, and the remainder arose from the retroperitoneum. They ranged in size from 2.1 to 32.0 cm and varied from tumors composed purely of rounded epithelioid cells to those composed of short fusiform cells set in a fine fibrillary collagenous background with some cases showing a mixed pattern. Tumors displayed variable amounts of stromal hyalinization, myxoid change, and cyst formation. The tumors expressed CD117 (c-kit receptor) (100%), CD34 (50%), neuron specific enolase (44%), smooth muscle actin (26%), desmin (4%), and S-100 protein (4%). Tumors were evaluated with respect to several parameters: size (<10 cm or >10 cm), cellularity (low or high), mitoses (0 to 2 per 50 high-power fields, >2 per 50 high-power fields), nuclear atypia (1 to 3+), cell type (epithelioid, spindled, or mixed), and necrosis (absent or present). These parameters were then evaluated in univariate and multivariate analysis with respect to adverse or nonadverse outcome, the former defined as metastasis or death from tumor. Follow up information was obtained for 31 patients (range, 4 to 84 months; median, 24 months). One patient presented with an adverse event and, therefore, was excluded from subsequent analysis. Twelve patients (39%) developed metastases or died of tumor. In univariate analyses, cellularity, mitotic activity (>2 per 50 high power fields), and necrosis were associated with statistically significant increases in the risk for adverse outcome. Despite the relatively small sample size, in a multivariable analysis mitotic activity (relative risk, 7.46; P = .09) and necrosis (relative risk, 3.75; P = .07) displayed trends toward independent predictive value. No association was noted between histologic pattern and outcome. Although only 39% of tumors behaved in a malignant fashion, this figure probably represents a conservative estimate because long-term follow-up (>5 years) was available for only a limited number of patients. Stratification of patients who have extragastrointestinal stromal tumor into those with 0 to 1 adverse histologic factors versus those with 2 to 3 offers the advantage of separating patients into two groups that have a markedly different risk for adverse outcome in the short term (0.02 events versus 0.54 events per person year; P < .001, respectively). Extragastrointestinal (soft tissue) stromal tumors are histologically and immunophenotypically similar to their gastrointestinal counterpart but have an aggressive course more akin to small intestinal than gastric stromal tumors. PMID- 10824932 TI - Association of a duodenal follicular lymphoma and hereditary nonpolyposis colorectal cancer. AB - Hereditary nonpolyposis colorectal cancer (HNPCC) is an inherited predisposition to colorectal and endometrial cancers caused by germline mutation of mismatch repair genes, with hMLH1 and hMSH2 underlying the majority of the cases. Although lymphoid tumors are the most common tumors in mouse models for HNPCC, lymphomas are almost never encountered in patients who have HNPCC, except in rare families with germline homozygous deletion of hMLH1. We report the case of a 53-year-old man who had a history of colon cancers related to constitutional hMLH1 mutation and who was diagnosed as having a duodenal follicular lymphoma This diagnosis was supported by IgH-BCL2 rearrangement and BCL2 immunoreactivity in tumor cells. The association of both of these possibly related rare diseases has never been reported. To clarify this relationship, we searched for hMLH1 expression and mismatch repair deficiency in the duodenal lymphoma. hMLH1 immunostaining was positive in lymphoid tumor cells, and no microsatellite instability was detected. In agreement with mouse models for HNPCC, these results suggest the involvement of alternative mechanisms to complete mismatch repair deficiency for lymphomagenesis in HNPCC syndrome. PMID- 10824933 TI - Collagenous gastritis: a case report, morphologic evaluation, and review. AB - Collagenous gastritis is rare; there are only four previous case reports. Histologic features seem to overlap with the other "collagenous enterocolitides"; however, pathologic criteria are not yet established for the diagnosis of collagenous gastritis. We describe an additional case of ostensible collagenous gastritis in a patient who initially presented with celiac sprue and subsequently developed colonic manifestations of mucosal ulcerative colitis. Endoscopic biopsies of the stomach revealed deposition of patchy, very thick bandlike subepithelial collagen in gastric antral mucosa, focal superficial epithelial degeneration, numerous intraepithelial lymphocytes, and a dense lamina propria lymphoplasmacytic infiltrate. Image analysis evaluation of gastric antral biopsies demonstrated a mean thickness of subepithelial collagen of 27.07 micron. Morphologic comparison was made with age-matched control groups of 10 patients who had normal gastric mucosal biopsies and 10 patients who had "chronic" gastritis, which revealed mean subepithelial collagen measures of 1.37 micron and 1.19 micron, respectively. We compared these morphologic findings with those of all previous case reports of collagenous gastritis and propose a pathologic definition based on the limited combined data. It seems that subepithelial collagen is dramatically thickened in reported cases of collagenous gastritis, with a cumulative mean measure of 36.9 micron. It is also apparent from this and previous reports that the thickened subepithelial collagen is accompanied by a chronic or chronic active gastritis and sometimes intraepithelial lymphocytes and surface epithelial damage. Recently described associations of lymphocytic gastritis, sprue, and lymphocytic colitis as well as collagenous and lymphocytic colitis suggest a common pathogenesis that empirically may include collagenous gastritis in the same disease spectrum. We propose that collagenous gastritis can be confidently identified by using analogous defined features of collagenous colitis: subepithelial collagen more than 10 micron in a patchy distribution, lamina propria lymphoplasmacytic infiltrates, intraepithelial lymphocytes, and surface epithelial damage. Collagenous gastritis also seems to have the same spectrum of associated clinical findings as collagenous colitis, including frequent coexistence of celiac sprue, watery diarrhea syndrome, and female predominance. PMID- 10824934 TI - Women and HIV--revisited ten years on. AB - This article compares how women and their relationship to HIV has changed since the original publication of this article in 1990. The number of women infected with HIV has continued to rise, but, in contrast, there have been few changes in their management. We review transmission risks and manifestations of the infection, and also discuss the issues faced by women with HIV. PMID- 10824935 TI - Rectal gonorrhoea in homosexual men: source of infection. AB - The objective of this retrospective study was to determine the possible source of infection in homosexual men with rectal gonorrhoea: the probable source of rectal gonorrhoea was identified in 46/155 cases. Although the urethra was the site of infection in 33 (72%) of these contacts, only pharyngeal gonorrhoea was identified in 9 (20%) men. In 25/26 cases, there was concordance in the auxo/serotypes of Neisseria gonorrhoeae between contacts with urethral gonorrhoea and the index men with rectal gonorrhoea. Eleven out of 12 pharyngeal isolates were of the same auxo/serotype as the index cases. This study supports the hypothesis that rectal gonorrhoea in homosexual men can be acquired from the oropharynx. Because infection at this site is an independent risk factor for acquisition of HIV, screening for rectal and pharyngeal gonorrhoea should be offered to men who have sex with men, even when there is no history of unprotected receptive anal intercourse. PMID- 10824936 TI - Enzywell recombinant enzyme immunoassay for the serological diagnosis of syphilis. AB - The aim of this study was to evaluate Enzywell TP, a new rapid enzyme immunoassay (EIA) that uses 2 recombinant Treponema pallidum antigens for the serological diagnosis of syphilis. Specificity was evaluated by screening 1055 unselected bloods requesting serological tests for syphilis in parallel with Enzywell TP and the Syphilis ICE EIA which is our standard screening test for syphilis. Sensitivity was evaluated using a panel of 159 known treponemal sera representing various stages of syphilis and 5 treponemal sera detected on screening. The specificity of Enzywell TP on initial and repeat testing (99.6% and 99.7% respectively) was similar to that of the Syphilis ICE test (99.8% and 99.9% respectively). The sensitivity of Enzywell TP (100%) was similar to that of Syphilis ICE (99.4%): both tests were significantly more sensitive (P=0.01) than the fluorescent antibody absorbed test (94.5%) but not the T. pallidum particle agglutination (TPPA) assay (99.4%). Both Enzywell TP and Syphilis ICE were positive with sera from 16 known HIV-infected patients who had been treated for syphilis many years previously (mean 9.4 years) confirming the value of these tests in excluding previous syphilis in HIV-infected individuals. We conclude that the Enzywell recombinant EIA is simple, rapid, highly sensitive and specific, and is a welcome addition to the range of currently available diagnostic tests for syphilis. PMID- 10824937 TI - Mycoplasma genitalium in non-gonococcal urethritis--a study in Swedish male STD patients. AB - Urethral swab specimens obtained from 101 men attending an STD clinic were examined for the presence of Mycoplasma genitalium by polymerase chain reaction (PCR) amplification. Fifty patients had non-gonococcal urethritis (NGU), and 51 patients were included as controls without urethritis. M. genitalium DNA was detected in 13 (26%) of the urethritis patients and in 5 (10%) of the control patients (P=0.06). No patient positive for M. genitalium had a simultaneous chlamydial infection. Thus, in the 36 patients with non-chlamydial NGU, the prevalence of M. genitalium infection was 36% (P=0.007 compared with controls). All patients with M. genitalium positive urethritis had a high grade urethritis defined as >10 polymorphonuclear cells per high power microscopical field. Compared with the control group, those with M. genitalium positive urethritis had more often had a history of urethritis than had those with chlamydial NGU or those with M. genitalium negative, non-chlamydial NGU. PMID- 10824938 TI - The ultrasound investigation of scrotal swelling. AB - The objectives of this study were to: (1) establish the causes of scrotal swelling in the hospital catchment population; (2) define the role of high frequency ultrasound examination in the management of scrotal swelling. A retrospective study of 582 patients who had high frequency ultrasound examination was carried out. Those requiring more information on perfusion had colour doppler examination. Forty-four per cent of examinations were performed for scrotal swelling. The cause of the scrotal swelling was mainly extratesticular (75% of all scrotal swellings), hydrocele being the commonest. Of the intratesticular causes, infection (50.8%) and tumour (20.6%) were the commonest. In conclusion ultrasound examination distinguishes extratesticular (almost always benign) from intratesticular (potentially malignant) causes of scrotal swelling. Infection, trauma and torsion mimic the ultrasound appearance of tumour as do rare benign entities. PMID- 10824939 TI - Uptake of HIV testing and treatment in an isolated population with access to free and universal healthcare. AB - This Western Australian study analysed clinical information from 344 people with AIDS, and experiential data from a sample of 36 people with AIDS to investigate 2 relevant issues: the timing of HIV testing and uptake and experiences of HIV drug treatment. We found that the proportion of people presenting with AIDS before being diagnosed with HIV infection ('AIDS-presenters') fell from 47% in 1985-1987 to 20% in 1991-1993. Since antiretroviral therapy (ART) became available in 1987, two-thirds of Western Australian people with AIDS who knew of their HIV infection prior to the development of AIDS have consistently elected to take recommended HIV treatment. Four phases of the treatment experience were identified. It is concluded that 'AIDS-presentation' is unlikely to reduce further and that the timing of testing and issues of treatment acceptance are only understood when the experiences of people with illness are examined using a patient-centred approach. PMID- 10824940 TI - Sexually transmitted infections and use of sexual health services among young Australian women: women's health Australia study. AB - Our objective was to examine associations between self-reported sexually transmitted infections (STIs) and sociodemographic, lifestyle, health status, health service use and quality of life factors among young Australian women; and their use of family planning and sexual health clinics and associations with health, demographic and psychosocial factors. The study sample comprised 14,762 women aged 18-23 years who participated in the mailed baseline survey for the Australian Longitudinal Study on Women's Health, conducted in 1996. The main outcome measures are self report of ever being diagnosed by a doctor with an STI, including chlamydia, genital herpes, genital warts or other STIs, and use of family planning and sexual health clinics. The self-reported incidence of STI was 1.7% for chlamydia, 1.1% genital herpes, 3.1% genital warts, and 2.1% other STIs. There was a large number of demographic, health behaviour, psychosocial and health service use factors significantly and independently associated with reports of having had each STI. Factors independently associated with use of family planning clinic included unemployment, current smoking, having had a Pap smear less than 2 years ago, not having ancillary health insurance, having consulted a hospital doctor and having higher stress and life events score. Factors independently associated with use of a sexual health clinic included younger age, lower occupation status, being a current or ex-smoker, being a binge drinker, having had a Pap smear, having consulted a hospital doctor, having poorer mental health and having higher life events score. This study reports interesting correlates of having an STI among young Australian women aged 18-23. The longitudinal nature of this study provides the opportunity to explore the long-term health and gynaecological outcomes of having STIs during young adulthood. PMID- 10824941 TI - Occurrence and treatment of Mycoplasma genitalium in patients visiting STD clinics in Sweden. AB - Two hundred and thirty-three men and 85 women visiting STD clinics in western Sweden between April 1997 and March 1998 were examined for Mycoplasma genitalium and Chlamydia trachomatis. The bacteria were identified by the polymerase chain reaction (PCR) technique. Three women (3.5%) and 18 men (7%) were positive for M. genitalium. Seventeen (14%) of the 115 men with urethritis were infected but only one of the men was without urethritis. After treatment with tetracyclines for 10 days, one woman and 8 of the 13 men still harboured M. genitalium. M. genitalium infected men did not have more life-time partners than other men visiting STD clinics. More men positive for M. genitalium gave a history of previous urethritis but the difference was not significant. PMID- 10824942 TI - Epidemiology of syphilis in Hungary between 1952 and 1996. AB - Trends in morbidity from syphilis in Hungary between 1952 and 1996 were analysed. The incidence of syphilis/100,000 inhabitants declined rapidly owing to the public health and therapeutic measures of the early 1950s (1952: total=73.6, early infections=60.2; 1962: total=13.7, early infections=8.7). After a temporary, slight increase until 1973 the number of reported syphilis cases declined continuously between 1978 and 1989 (1989: total=0.9, early infections=0.84). In 1994 a marked increase occurred when compared with 1993 (1993: total=early: 1.4. 1994: total=2.3, early infections=2.2). Incidence trends were statistically analysed using Chi-square test and linear regression. Chi square analysis showed that the changes in the incidence of total and early syphilis are significant (P<0.00001) comparing the time intervals 1952-1962 with 1962-1966 and 1975-1979 with 1988-1992. The same trends were found using the linear regression test, except for the time interval of 1960-1973. PMID- 10824943 TI - Amyl nitrite induced acute haemolytic anaemia in HIV-antibody positive man. AB - Volatile nitrites (amyl and butyl) are popular recreational drugs, especially in the homosexual population. Haemolytic anaemia is a rare complication of nitrite inhalation and occurs when the reducing capacity of the red cell enzymes is overcome by the oxidizing effect of the nitrite. We describe here a patient with HIV infection who developed a profound haemolytic anaemia after repeated inhalation of large quantities of amyl nitrite. PMID- 10824944 TI - Late onset hepatitis and prolonged deterioration in hepatic function associated with nevirapine therapy. AB - The aetiology of hepatic dysfunction in patients with HIV infection is multifactorial. Re-activation of hepatitis C infection, drug toxicity, and opportunistic infections are all potential causes. Nevirapine (NVP) is a non nucleoside reverse transcriptase inhibitor used as part of combination antiretroviral therapy for the treatment of HIV infection. It is associated with a significant incidence of hepatotoxicity, usually occurring in the initial month of therapy. We report the case of a 49-year-old man who developed NVP-induced prolonged hepatotoxicity 5 months after commencing antiretroviral therapy. PMID- 10824945 TI - Number of consultants in genitourinary medicine (GUM) clinics in England, Scotland and Wales. PMID- 10824946 TI - Influence of selection for increased body weight, egg production, and shank width on the length of the incubation period of turkeys. AB - An experiment was designed to study the influence of selection for increased 16 wk BW (F line), egg production (E line), and shank width (FL line) in turkeys on length of the incubation period and to estimate the influence of adult BW (BW50) and egg weight on the incubation period. The base populations for the F, FL, and E lines were randombred control (RBC)2, F, and RBC1, respectively. An additional RBC (RBC3) was also included in the study. The experimental turkey lines differed in length of the incubation period in all trials, indicating genetic differences exist. The E line, in general, had a longer incubation period than the RBC1 line, due to an increase in the time required for external pipping. The FL line had a longer period of incubation than the F line. Differences in the length of the incubation period between the F and RBC2 lines were inconsistent. The F line had a shorter incubation period than the RBC2 line in Trial 1, whereas there was no difference between lines in Trial 2. In Trial 3, the comparison of the F and RBC2 lines was influenced by treatment of the eggs prior to setting. When the eggs were held at room temperature for 29 h prior to setting, the F line had a longer incubation period than the RBC2 line, whereas when the eggs were set directly from the egg cooler, there was no line difference. The line differences in length of the incubation period could not be explained by line differences in egg weight based on a comparison of line means for these traits. Within lines, the regression of length of the incubation period on egg weight was positive and significantly different from zero in three of the six lines. Overall regression of hatching time on egg weight and BW50 was positive for egg weight and negative for BW50. However, only 4% of the variation in the length of the incubation period could be explained by variation in egg weight and BW50. Within lines, the regression coefficient of length of the incubation period on BW50 was not significantly different from zero in any line but the sign was negative in five of six lines. PMID- 10824947 TI - Analysis of disease resistance-associated parameters in broiler chickens challenged with Eimeria maxima. AB - To determine an optimal dose for coccidial inoculation and to evaluate genetic resistance or susceptibility in individual chickens, broilers were inoculated with four different doses of Eimeria maxima oocysts. Body weight gain, fecal oocyst shedding, concentrations of plasma NO2- + NO3-, carotenoid, and interferon gamma were measured at two different time periods postinfection. The results showed significant dose and sex effects on most parameters and interaction between dose and sex in some parameters. The dose effects were generally linear; however, some significant quadratic effects were also observed. The measurements from chickens inoculated with 10(4) oocysts displayed the highest correlation coefficients among oocyst shedding, body weight gain, and concentrations of carotenoid and NO2- + NO3-. An infection index, calculated from the correlated parameters, displayed high correlation coefficients with the parameters. The infection index may be a better parameter for evaluating individual genetic resistance against coccidial infection. PMID- 10824948 TI - Microsatellite polymorphism between and within broiler populations. AB - Two independent broiler chicken populations were genotyped with microsatellite markers to determine genetic polymorphisms within and among broiler populations. Birds were genotyped with primers from the US Poultry Genome Mapping Kits 1 and 2. The 59 primer sets selected for this study provided wide genomic coverage. All 59 primer sets amplified a polymerase chain reaction product in Population L, whereas 57 primer sets produced a product in Population C. The average allele number per line per microsatellite was 2.8 and 2.9 for Populations L and C, respectively. Considering the 57 primer pairs generating product in both lines, 72.3% of the total alleles were unique to one or the other population. This study illustrates the high polymorphism level in broiler populations of microsatellites amplified from primers developed from Red Jungle Fowl or White Leghorn sequences. PMID- 10824949 TI - Effects of dietary fat type and level on broiler breeder performance. AB - The effects of added dietary fat type and level on broiler breeder performance were evaluated in two trials. In Trial 1, diets included no added fat (NAF) or 3.0% added poultry fat (PF) for individual bird energy intakes of 430 and 467 kcal/d at peak production (PCD), or 1.5% PF or 3.0% corn oil (CO) at 449 PCD. In Trial 2, diets contained NAF, 1.5 or 3.0% added CO or PF, or 3.0% lard (LA) for peak energy intakes of 463 PCD. All diets in Trial 2 were isocaloric and isonitrogenous. Feeding of experimental diets was initiated when breeders were 22 wk old in both trials, and hen BW was measured at various weeks between 22 and 47 wk in Trial 1 and between 27 and 65 wk in Trial 2. Egg production, egg weight (EW), eggshell quality, and hatchability were recorded at various weeks between 24 and 48 wk in Trial 1 and between 27 and 64 wk in Trial 2. Body weights and EW increased progressively, and eggshell quality fluctuated variably with age. Egg production peaked between Weeks 32 and 35, and hatchability was significantly depressed at Week 63. Egg weight and hatchability were not affected by dietary treatment; however, BW increased with energy level in the diet between Weeks 27 and 47. Body weight was also higher in birds fed 1.5 or 3.0% added fat compared with NAF and was higher in birds fed LA compared with CO at the 3.0% level. Egg specific gravity and shell weight per unit of surface area at Week 41 were increased by feeding low or high energy compared with moderate energy, and relative eggshell conductance was increased by using moderate compared with high energy diets and by feeding 1.5% PF compared with 3.0% PF. Egg specific gravity was increased by PF when compared with CO across fat level at Weeks 26 and 47 and by LA when compared with PF across fat level and breeder age. These data suggest that certain dietary energy and fat regimens may affect BW and eggshell quality without associated effects on EW and hatchability at various breeder ages. PMID- 10824950 TI - Assessment of demand for a job placement service for the Poultry Science Association. AB - A placement service for the membership of the Poultry Science Association (PSA) is a possible future consideration. This service should provide a formal national platform that could aid membership and their employers in position services. A survey was conducted with the objective of determining the interest of the membership in such a placement service and the maximum expense that members would be willing to pay for the service. Ten questions in an undisguised questionnaire format, covering demographic, historical, and interest points was devised with survey questions designed for a forced response on most of the questions. A total of 2,050 surveys was mailed to PSA members in the US, including 1,850 regular members and 200 student members; 518 responded to the survey. A majority of all age groups favored the construction of a placement service, but the respondents' willingness to use the service depended upon age, education, and work experience and whether or not they believed a placement service was needed. Sources of information used in finding a position varied with age; trade journals and publications were the most frequent sources for the responses. It appears that the membership likes the idea of a placement service, is willing to pay something for it, and would use it if it were available. PMID- 10824951 TI - Struggling behavior in shackled male and female broiler chickens. AB - Anecdotal evidence suggests that the struggling behavior of shackled broiler chickens may be positively related to compression of the shank and the probable associated discomfort: birds with large shanks tend to struggle more violently than do those with smaller shanks. Males are generally heavier and have thicker shanks than females. Therefore, we tested the hypothesis that, because the leg gaps of shackles are fixed in size, male broilers would struggle more than females. At 42 d of age, 264 floor-reared broilers were cooped in groups of 12 (six males and six females) and were transported from the university farm to the abattoir. Eighty of these served as test birds (n = 40/sex) and were shackled on a moving processing line with a bird of randomly selected sex on either side. Upon shackling, the latencies to struggle, numbers of struggling bouts, and total time spent struggling were recorded during a 1-min test period. Subsequently, the BW and circumference of the right shank (CRS) of each test bird were measured. Male birds were heavier and had thicker shanks than females (both P < 0.0001); they also struggled sooner (P < 0.01) and longer (P < 0.008). When data from males and females were pooled, CRS was negatively correlated with latency to struggle (r = -0.30; P < 0.006) and positively associated with SB (r = 0.23; P < 0.04) and total time spent struggling (r = 0.23; P < 0.04). However, there were no detectable correlations within sex. Body weight was not significantly correlated with any of the struggling behavior measures. Although other gender related factors may be influential, an interpretation of our findings based on sex differences in CRS seems the most parsimonious. We conclude that use of shackles of fixed leg-gap size may contribute to increased struggling behavior in male broilers. PMID- 10824952 TI - D-xylose absorption as a measurement of malabsorption in poult enteritis and mortality syndrome. AB - Severe wasting of body tissues, diarrhea, high morbidity and mortality, and stunting are all characteristics of poult enteritis and mortality syndrome (PEMS). The wasting of musculature and loss of nearly all adipose tissue suggested that even though the PEMS-infected poults were eating some feed, nutrient intake was not sufficient to meet body requirements for maintenance and growth. Because epithelial cells in the gastrointestinal tract appeared to be a target of the undefined etiological agent (or agents) that causes PEMS, a study was conducted in which PEMS-infected poults were evaluated for malabsorption through 3 wk of age. D-Xylose, a poorly metabolized pentose, was given per os as a bolus, and blood samples were obtained from the ulnar vein in the wing of control and PEMS-infected poults over a 3-h period to estimate intestinal absorption. D-Xylose absorption in control poults peaked 30 to 60 min after the oral treatment, similar to results reported earlier. The PEMS-infected poults did not show a peak in absorption. The PEMS-infected poults showed significant delays in D-xylose absorption at 4, 7, and 11 d after PEMS challenge. The severe malabsorption and metabolic deficiency problem associated with PEMS was postulated to be a direct effect of the undefined infectious agent or agents that cause the disease. PMID- 10824953 TI - Influence of BioChrome on the response of metabolic hormones in PEMS-infected poults. AB - Poult enteritis and mortality syndrome (PEMS), a disease that affects turkeys between 7 and 28 d of age, causes a severe inflammation of the intestinal tract and is characterized in poults by severe diarrhea, high morbidity, mortality, and stunting. The PEMS-associated mortality and growth depression is related to malabsorption and decreased metabolic activity caused, in part, by a possible insulin deficiency or insensitivity. Insulin receptors are stimulated by the glucose tolerance factor (GTF) that incorporates Cr. Body Cr deficiency can be exacerbated by dietary deficiency and by increased excretion due to stress associated with a diarrheal disease such as PEMS. BioChrome (BC) contains natural, preformed GTF, the bioactive form of Cr. Experiments were conducted in which BC was blended into poult starter feed at 400 ppb during the first 21 d posthatch. Body weights were determined at 1, 7, 14, and 21 d of age, and weekly feed conversions were calculated for each treatment group (control, BC, PEMS, and BC+PEMS). At 6 d post-hatch, each PEMS-designated poult was given a 0.1-mL oral gavage of a 10% suspension of feces from PEMS-infected poults. Blood samples were taken via cardiac puncture from four birds per treatment group at 7, 10, 14, 17, and 21 d of age. Radioimmunoassays were conducted for plasma insulin, glucagon, thyroxine (T4), and triiodothyronine (T3). Plasma insulin levels were depressed in PEMS-infected poults from Days 10 through 17, but plasma glucagon levels in the PEMS-infected poults were significantly elevated at 14 and 17 d, after which they returned to control levels in both of the PEMS-infected groups. The T3 and T4 levels were depressed through Day 21 in PEMS-infected poults, but with BC treatment these blood hormone levels rebounded by Day 21. Body weights of PEMS infected poults were increased significantly by the BC treatment but not to the level of noninfected controls. PMID- 10824954 TI - The effect of environmental temperature on growth and feed conversion of broilers to 21 days of age. AB - Temperature during rearing is a major factor in production efficiency for broilers. The objective of this research was to determine the effect of five temperature regimens on the growth, feed conversion, and mortality of broilers to 21 d of age. This information is needed to select the most profitable conditions for broiler production. Three trials were conducted in 10 environmental chambers; five chambers were stocked with males, and five were stocked with females. The chambers were initially set at 28, 29, 30, 31, and 32 C followed by daily reductions of 0.3 C for each beginning temperature. The dewpoint was set at 15.7 C throughout the experiments. Lighting was continuous, and feed and water were provided ad libitum. Weight gain and feed conversion were determined at 7, 14, and 21 d, and mortality was recorded as it occurred. Temperature did not affect weight gain or feed conversion for any period, but males were superior to females for these parameters for the 7- to 14-d and 14- to 21-d periods. Temperature affected mortality; most of the deaths occurred during the 0- to 7-d period and at the lowest temperature treatment. These results suggest that mortality is the best indicator of the effect of rearing temperature on performance. PMID- 10824955 TI - The effect of vitamin D3 on resistance to stress-related infection in an experimental model of turkey osteomyelitis complex. AB - Male turkeys immunosuppressed by injection with dexamethasone (DEX) were given supplemental vitamin D3 in their drinking water in two experiments. In Experiment 1, vitamin D3 was supplemented at a dosage of either 2,064 IU/kg (low level) or 4,128 IU/kg (high level) in drinking water provided ad libitum only from Days 1 through 5 after hatch. In Experiment 2, vitamin D3 was provided at the low dosage for the first 5 d after hatch, followed by treatment with the high dosage for 12 h before and 12 h after each stressful event, which included weekly weighings and two DEX treatments. In both experiments, at 5 wk of age half of the birds were given intramuscular injections of 2 mg/kg DEX on 3 alternating d. In Experiment 1, 100 cfu of Escherichia coli was inoculated into the left thoracic airsac at the time of the third DEX injection. All mortalities were examined, and 10 birds per pen were necropsied 2 wk after treatment and examined for lesions of airsacculitis and turkey osteomyelitis complex (TOC). Four birds per pen were bled before necropsy, and white blood cell total counts, differential white blood cell counts, and clinical chemistry values were determined. In Experiment 2, healthy surviving birds were grown for an additional 5-wk period, after which the DEX-treated birds were given a second series of DEX injections and were bled and necropsied 2 wk later. There were no significant effects of vitamin D3 treatment in combined general linear models analysis of Experiment 1; however, when birds not treated with DEX or E. coli were compared with those treated with both DEX and E. coli, supplementation with the low level of vitamin D3 significantly decreased TOC incidence. There were no significant effects of vitamin D3 treatment in birds treated with DEX at 5 wk of age in Experiment 2. However, when surviving birds were given a second DEX treatment at 12 wk, vitamin D3 treatment resulted in significantly lower incidence of mortality, TOC, green liver, isolation of bacteria from tissues, and lower airsacculitis scores and heterophil to-lymphocyte ratios than controls. Vitamin D3 also improved BW, relative weights of the liver and heart, and serum levels of glucose and alanine aminotransferase (ALT) of birds receiving two treatments with DEX. The ability of vitamin D3 supplementation to protect turkeys from the immunosuppressive effects of multiple DEX treatments emphasizes the role of vitamin D3 as a prohormone that affects health and disease resistance in turkeys. PMID- 10824956 TI - Ascorbic acid supplementation improved antibody response to infectious bursal disease vaccination in chickens. AB - The purpose of the present study was to determine if supplementation of ascorbic acid (AA) to the diet would have a beneficial effect on infectious bursal disease (IBD) vaccination of chickens for protection against infectious bursal disease virus (IBDV) infection. Two hundred forty specific pathogen-free (SPF) chickens were divided into eight experimental groups. A 2 x 2 x 2 factorial arrangement in a completely randomized design was used; AA supplementation at 1,000 ppm in the diet, vaccination, and challenge were the main effects. Prior to challenge and 10 d after challenge, serum AA concentration, serum corticosterone concentration, ELISA antibody titer to IBDV, body weight, bursa-to-body weight (B:B) ratio, and bursal histological score (BHS) were determined. Nonvaccinated chickens fed a diet supplemented with AA did not exhibit clinical signs or mortality following challenge, whereas AA-unsupplemented counterparts had 100% cumulative morbidity and 30% cumulative mortality. Serum AA levels of AA-supplemented and vaccinated chickens were significantly (P < 0.05) higher than AA-unsupplemented and vaccinated chickens. Fourteen days following vaccination, significantly (P < 0.05) higher ELISA titers to IBDV were observed in vaccinated chickens supplemented with AA as compared to AA-unsupplemented counterparts. Ascorbic acid supplemented chickens, especially those also vaccinated, had higher body weight gains as compared to the AA-unsupplemented chickens. Ascorbic acid-supplemented chickens challenged with IBDV did not show any clinical signs or mortality. The results suggest that supplementation of AA at 1,000 ppm in the diet has beneficial effects on antibody response to IBD vaccination and body weight gain. PMID- 10824957 TI - Preventive effect of partially hydrolyzed guar gum on infection of Salmonella enteritidis in young and laying hens. AB - The preventive effect of partially hydrolyzed guar gum (PHGG) on the colonization of Salmonella enteritidis (SE) in young and laying hens was investigated. The effects of feed supplemented with 0.025, 0.05, and 0.1% PHGG was examined on young hens orally infected with SE. The incidence of SE in organs was decreased, the excretion of SE into feces was increased, and the agglutinating antibody titer to SE in serum was decreased by the administration of PHGG to young hens. In particular, feed supplemented with 0.025% PHGG was the most effective. It was also shown that feed supplemented with 0.025% PHGG increased the number of Bifidobacterium spp. and Lactobacillus spp., the most numerous intestinal bacteria in the cecum of young hen. The effect of the excretion of SE via feces was also observed in an experiment using laying hens. The incidence of SE on the surface of the eggshell and in egg white and egg yolk was also decreased when the feed of laying hens was supplemented with 0.025% PHGG. These results show that the administration of feed supplemented with PHGG can prevent the colonization of SE in young and laying hens, which, in turn, could be related to improvement in the balance of intestinal microflora. PMID- 10824958 TI - Effects of breeder age and dietary fat source and level on broiler hatching egg characteristics. AB - The effects of breeder age and added dietary fat source and level on broiler hatching egg characteristics were evaluated. Diets included no added fat (NAF) or 3.0% added poultry fat (PF) for peak energy intakes of 430 and 467 kcal/hen-day (PCD) or 1.5% PF or 3.0% corn oil (CO) at 449 PCD. As added dietary fat was changed from CO to PF, the percentage of unsaturated dietary fatty acids, including linoleic acid, decreased. Feeding of experimental diets was initiated when breeders were 22 wk old. Total fresh egg weight; eggshell weight; percentages of yolk (PYK), albumen (PAB), and eggshell (PSHL) weights; and yolk:albumen ratio were measured at various weeks between 26 and 47 wk of age. Egg weight increased progressively with hen age. Significant increases in yolk:albumen ratio occurred between Weeks 26 and 31 and between Weeks 31 and 35. Low (430 PCD) dietary energy levels significantly reduced PYK at 35 wk and increased PAB across breeder age. Eggshell weight was lower in birds fed moderate (449 PCD) compared to low energy levels at Week 26, moderate compared to high (467 PCD) energy levels at Week 41, and PF compared to CO across fat level at Week 31. At Weeks 31 and 41, PSHL was increased by the use of 3.0% PF compared to 1.5%, and PSHL was increased at Weeks 26 and 41 by using added PF compared to CO across fat level. Increased dietary energy decreased PAB and the use of added dietary CO rather than PF decreased PSHL in broiler breeders between 26 and 47 wk of age. PMID- 10824959 TI - Spatial distribution of cannibalism mortalities in commercial laying hens. AB - The distribution of cannibalism cases in a flock of 19,776 Babcock White Leghorns was monitored from 21 to 54 wk of age. The hens were kept in a single-floor house consisting of four banks of two-deck stair-step cages. Each of the 4,944 cages held four hens at a density of 152 cm(2) (60 inches(2)) per hen. Each cage was assigned a number from 1 to 4,944, and each dead bird was tagged according to its cage of origin. Dead birds were collected daily, kept in a freezer, and necropsied weekly. Farm personnel routinely transferred a live hen from an end cage to a cage where a mortality had occurred. The cause of death, age, cage number, and cage location were recorded for each dead hen. Of the 1,173 hens that died during the study period, 253 (21.6%) died from egg peritonitis, 184 (15.7%) from hypocalcemia, 167 (14.1%) from cannibalism, 164 (14%) from neoplastic disease, and the rest from various other causes. Cannibalism cases were analyzed statistically for clustering. Cannibalism was defined as death from tissue trauma and hemorrhage inflicted by cage mates. A spatial analysis showed that cannibalism is not a random event but one that occurs in clusters. The incidence of cannibalism was also found to be significantly higher on the top rows of cages as compared with the bottom rows. PMID- 10824960 TI - Comparison of genetically engineered microbial and plant phytase for young broilers. AB - This study was conducted to compare the efficacy of genetically engineered microbial (Natuphos) and plant (Phytaseed) phytase for enhancing the utilization of phytate P in corn-soybean meal-based diets fed to young broilers and to evaluate the safety of Phytaseed phytase. Three levels of each of the two sources of phytase (250, 500, and 2,500 U/kg of diet) were added to a corn-soybean meal basal diet containing 0.46% total P, 0.21% nonphytate P, and 0.92% Ca. There were eight cages per treatment (eight birds per cage for Weeks 2 to 3 and seven birds for Weeks 4 to 5), except for the basal diet without added phytase that had 16 cages. Cage BW and feed consumption were recorded weekly. During Week 5, cage excreta samples were collected for determination of apparent retention coefficients of DM, Ca, and P. At the end of Week 5, all birds were killed, and the left and right toes were removed for determination of toe ash weight and percentage. Forty birds (one per cage from the diet without added phytase and diets with 500 or 2,500 U phytase/kg from both sources) were randomly selected for gross necropsy and histologic evaluation of liver, kidney, and bone tissues. Addition of both sources of phytase resulted in similar increases (P < 0.05) of BW gain; feed intake; gain:feed; apparent retention of DM, P and Ca; and toe measurements. Phosphorus excretion decreased as phytase addition increased. No significant abnormalities were seen in any of the 40 broilers necropsied. Further, the fit of a nonlinear function revealed that most measurements reached a plateau at 2,500 U/kg. Based on performance, bone characteristics, and retention of P, Ca, and DM of young broilers, the efficacy of Phytaseed phytase was similar to that of Natuphos phytase for enhancing the utilization of phytate P in corn-soybean meal-based diets. General necropsy and histologic examination of liver, kidney, and tibial tissues revealed no adverse effects of phytase source or level. PMID- 10824961 TI - Recovery responses of chick intestinal villus morphology to different refeeding procedures. AB - Recovery responses of intestinal villus height and fine structure on the villus apical surface to different refeeding procedures were compared at refeeding 3 or 24 h after 3-d of feed withdrawal from chicks. After 3-d of fasting, 45-d-old male White Leghorn chicks (Gallus gallus domesticus) were refed rice bran (RB) (CP, 14.8%; ME, 3,170 kcal/kg), conventional grower mash diet (CG) (CP, 18.2%; ME, 2,853 kcal/kg), or ground CG (GG). During the experimental period, birds were given access to diets and water ad libitum. After the end of each experimental period, the duodenum was fixed and examined by light and scanning electron microscopy for morphological changes in the villi. Duodenal villus heights of chicks fasted for 3 d were significantly decreased compared with control chicks fed ad libitum. Villus heights were significantly increased after a 3-h refeeding, and the villi of the GG-refed group were significantly higher than RB and CG. When compared with villus heights at the 3-h refeeding, the RB-refed group showed no changes after a 24-h refeeding, but the CG and GG groups had significantly increased villi heights. Villi heights in GG groups were significantly higher than in the CG groups. Dietary effects on cell area and cell mitosis numbers were similar to those observed for villus height. The surface of the duodenal villi apices of control birds fed ad libitum revealed a clear cell outline, cell protuberances, and cell extrusion into the lumen. After 3-d fasting, cell outlines became faint, and protuberances and extrusion of cells disappeared. After refeeding for 3 h, clear cell outlines were again apparent in all groups. In GG-refed chicks, larger cell outlines and protuberated cells were found as conspicuous morphological features. Similar observations were made at the 24-h refeeding. These morphological findings suggest that chickens that were on feed withdrawal benefit from ad libitum refeeding of a powdered diet that is nutritionally complete for rapid recovery of digestive function. PMID- 10824962 TI - Dietary levels of chia: influence on yolk cholesterol, lipid content and fatty acid composition for two strains of hens. AB - Four hundred fifty H&N laying hens, half white and half brown, were fed for 90 d to compare a control diet to diets containing 7, 14, 21, and 28% chia (Salvia hispanica L.) seed. Cholesterol content, total fat content, and fatty acid composition of the yolks were determined 30, 43, 58, 72, and 90 d from the start of the trial. Significantly less cholesterol was found in the egg yolks produced by the hens fed the diets with 14, 21, and 28% chia compared with the control, except at Day 90. Palmitic fatty acid content and total saturated fatty acid content decreased as chia percentage increased and as the trial progressed. Total omega-3 fatty acid content was significantly greater (P < 0.05) for both strains for all chia diets compared with the control diet. Total polyunsaturated fatty acid (PUFA) content of the yolks from the chia diets was significantly greater (P < 0.05) than from the control diet. Generally, total PUFA content tended to be highest in the yolks of the white hens. PMID- 10824963 TI - Evaluation of tryptophan requirement of the commercial layer by using a corn soybean meal basal diet. AB - An experiment was conduced with Hy-Line W36 hens to evaluate the requirement for Trp in a corn-soybean meal diet. Seven experimental diets were fed with Trp levels of 0.12, 0.13, 0.14, 0.15, 0.16, 0.17 and 0.18%. Supplemental amino acids (AA) were added to all diets to ensure that Trp was the first-limiting AA. A positive control diet (0.20% Trp) with Met supplementation was fed that had previously been shown to support maximum performance. Egg production (EP), egg weight (EW), and egg content (EC) were significantly increased by the addition of Trp to the basal diet. Broken-line regression indicated the Trp requirement for EP and EC was 139.8 and 149.0 mg per hen/d, respectively, for EP and EC when hens had a daily EC of 45.4 g per hen/d. PMID- 10824964 TI - Effect of dietary concentration of xylitol on growth in male broiler chicks during immunological stress. AB - Two experiments were conducted to determine the effect of dietary xylitol concentration on growth performance, plasma (alpha1 acid glycoprotein (AGP), nitrite, and Fe concentration in male broiler chicks during immunological stress. Ten-day-old chicks were fed a corn-soybean diet containing 15% glucose and 6% xylitol or 15% xylitol with identical metabolizable energy and crude protein content for 12 d in Experiment 1. In Experiment 2, 12-d-old chicks were fed either the 15% glucose or 6% xylitol diet for 7 d. During the final 6 d of each experiment, half of the birds fed each diet were injected intraperitoneally with Escherichia coli lipopolysaccharide (LPS; 0127:B8) on Days 1, 3, and 5 and with Sephadex-G50 superfine on Days 2 and 4 to stimulate the immune system. The xylitol diets partially prevented reductions in body weight gain, feed intake, and feed efficiency caused by LPS and Sephadex injections, but the glucose diet did not. The injections of LPS and Sephadex increased plasma AGP and nitrite concentrations. Plasma AGP concentration on Days 2 and 6 in chicks fed the xylitol diets did not differ from that of chicks fed the glucose diet in both experiments. Nitric oxide production estimated by plasma nitrite concentration following immunological stress did not differ due to dietary treatments in Experiment 2. The LPS and Sephadex resulted in decreased plasma Fe concentration on Day 6 in Experiment 1 in chicks fed glucose but not xylitol. These results indicate that a beneficial effect of dietary xylitol on growth is obtained with 6% xylitol given to chicks 1 d before stimulating the immune system. PMID- 10824965 TI - Nonphytate phosphorus requirement of laying hens with and without phytase on a phase feeding program. AB - An experiment was conducted to reevaluate the nonphytate P (NPP) requirement of laying hens with and without phytase. The experiment involved 12 treatments in a 6 x 2 factorial design. The hens of the control group (T1) were fed a sequence of 0.40-0.35-0.30% NPP during 30 to 42, 42 to 54, and 54 to 66 wk, respectively. The NPP was reduced in increments of 0.05% in T2 to T6. The hens of T7 to T12 were fed NPP regimens similar to T1 to T6 but with 300 units phytase/kg diet. Two digestion trials were conducted during 42 and 66 wk, and nitrogen, phytate, and total P retention were determined. In the absence of phytase, production traits were not different for hens fed a NPP regimen of 0.25-0.20-0.15% than for the unsupplemented phytase control group for the entire experiment (P > 0.05). However, production traits were inferior for hens fed the lower NPP regimens (P < 0.05). In the presence of phytase, production traits were not different for hens fed the lowest NPP regimen (0.15-0.10-0.10%) than for the unsupplemented phytase control for the entire experiment (P > 0.05). The overall effect of NPP regimens and phytase on specific gravity for the entire experiment was significant. Specific gravity was greater for hens fed the regimens with less NPP than for hens fed regimens with greater NPP, and phytase had an adverse effect on specific gravity. However, specific gravity of hens fed the lowest NPP regimen with phytase was not significantly different from the unsupplemented phytase control group. The overall effect of phytase on phytate P retention was significant during digestion trials; phytase increased phytate P retention by about 15%. Daily total P excretion was 34 to 47% less for hens fed the lowest NPP regimen with phytase than for the unsupplemented phytase control group. Significant interactions existed for a number of traits and are discussed in the text. PMID- 10824966 TI - Efficacy of phase-feeding in supporting growth performance of broiler chicks during the starter and finisher phases. AB - A feeding regimen has been developed that uses regression equations to predict amino acid requirements over time. Phase-feeding (PF) of broilers was tested to evaluate its efficacy compared with feeding broilers NRC or Illinois ideal chick protein (IICP) recommendations. In Experiment 1, NRC or IICP requirements for lysine, sulfur amino acids, and threonine were fed from 0 to 21 d, whereas PF was tested using a series of three diets (0 to 7, 7 to 14, and 14 to 21 d). No differences (P > 0.05) in weight gain, feed intake, feed efficiency, digestible amino acid intake, or gain per unit digestible amino acid intake were noted among chicks fed NRC, IICP, or PF diets. In Experiment 2, NRC or IICP requirements were fed from 40 to 61 d, whereas PF was tested using a series of three diets (40 to 47, 47 to 54, and 54 to 61 d). No differences (P > 0.05) in weight gain or feed intake were observed, but the feed efficiency of birds fed the IICP diet was decreased (P < 0.05). The IICP and PF diets resulted in decreased (P < 0.05) digestible lysine and threonine intake; gain per unit digestible lysine and threonine intake was increased (P < 0.05) by PF. No differences (P < 0.05) in breast meat, wing, or leg yield were noted among treatments. Economic analysis indicated that PF may facilitate reduced dietary costs without sacrificing growth performance or carcass yield. PMID- 10824967 TI - Use of a sperm analyzer for evaluating broiler breeder males. 2. Selection of young broiler breeder roosters for the sperm quality index increases fertile egg production. AB - Previous research has shown that the sperm quality index (SQI) of rooster semen is indicative of overall semen quality. The objectives of the present experiments were to determine the correlation of the SQI with semen characteristics and fertility and to determine if selection of young males for the SQI would improve fertility. In Experiment 1 semen was collected from 35 Peterson males and was analyzed individually for sperm concentration and viability. To determine fertility, 100 microL of diluted semen was inseminated into 10 hens for each rooster. Positive correlations of the SQI with total and live sperm concentrations as well as fertility were found. A negative correlation of the SQI with the percentage of dead sperm was observed. In Experiment 2, four semen samples were collected at 2- to 3-d intervals from each of 142, 27-wk-old Peterson roosters to determine their SQI. Males were then allocated to six treatment groups based on their average SQI readings as follows: 0 to 150, 151 to 200, 201 to 250, 251 to 300, 301 to 350, and >350. For each SQI group, semen was collected weekly for 8 wk, pooled, and used at a rate of 50 microL/hen to inseminate 40 hens. The percentage of fertilized eggs increased linearly across the SQI groups, from a minimum of 65% for the 0 to 150 SQI group to a maximum of 98% for the >350 SQI group. The SQI groups of 301 to 350 and >350 produced the slowest decline in fertility over days postinsemination. Therefore, selection of males for the SQI at an early age appears to improve flock fertility. PMID- 10824968 TI - Effect of delayed step-up lighting on plasma luteinizing hormone and reproductive function in broiler breeders. AB - The aim of the study was to establish, in contemporary broiler breeders, whether delayed photostimulation at 313 d results in a reproductive response similar to that after photostimulation at 134 d (standard practice). The standard lighting program was compared with a novel program in which daily hours of light were reduced to 3 h during rearing and kept at 3 h until photostimulation at 264 d (8 h) or at 313 d (16 h). This experiment was done with hens fed ad libitum or feed restricted hens. In photostimulated and nonphotostimulated hens, feed restriction delayed the onset of egg production and enhanced the subsequent rate of laying. Standard photostimulation advanced the onset of lay and increased the subsequent rate of lay in hens fed ad libitum and feed-restricted hens. Delayed photostimulation of hens did not impair the photoinduced increase in the concentration of plasma luteinizing hormone (LH) or egg production. Delayed photostimulation in cockerels failed to stimulate LH secretion. Unexpectedly, for feed-restricted hens, transfer from 3 to 8 h light/d at 264 d resulted in an increased in plasma LH and increased egg production. A similar increase in plasma LH was observed for cockerels subjected to the same lighting treatment. We concluded that, in broiler breeder hens, the reproductive response to photostimulation is not impaired if photostimulation is delayed for up to 313 d. Cockerels may not respond well to delayed photostimulation. PMID- 10824969 TI - The relationship of raw broiler breast meat color and pH to cooked meat color and pH. AB - Three replicate trials were conducted to determine the influence of raw breast meat color and pH on subsequent cooked meat color and pH. In each trial, approximately 50 breast fillets were obtained from a commercial processing plant based on being either normal, lighter than normal, or darker than normal. Color (L* = lightness, a* = redness, and b* = yellowness) of each fillet was determined in triplicate on the underside surface of the fillet (to avoid scalding effects), and the pH was determined on a tissue sample removed from the posterior portion of each fillet. Fillets were then cooked in steam at 98 C for 20 min and cooled to room temperature, and a second sample was removed from the posterior section for cooked meat pH. Cooked meat color was measured on an exposed surface, to avoid cooking-related discoloration. The data were subjected to linear regression analysis to determine the relationship between raw and cooked values. Results indicated a significant linear relationship between raw and cooked values for each color parameter as well as pH. Model R2 values were 0.43, 0.40, 0.64, and 0.78 for L*, a*, b*, and pH, respectively. There were also significant linear relationships between raw meat L* and raw muscle pH (R2 = 0.59) as well as cooked meat L* and raw meat pH (R2 = 0.36). These results indicate that raw breast meat color and pH affect cooked breast meat color and pH but that cooking reduces the degree of color variation. Moreover, cooked meat lightness is more closely associated with raw breast meat pH than with cooked meat pH. PMID- 10824970 TI - The use of halothane gas to identify turkeys prone to developing pale, exudative meat when transported before slaughter. AB - Halothane screening has been used in the swine industry to identify animals susceptible to stress and prone to developing pale, soft, exudative (PSE) meat. This study evaluated the ability of halothane to identify stress-susceptible turkeys prone to developing PSE meat when reared to market age and transported before slaughter. Male Nicholas turkeys (n = 1,286) were exposed to 3% halothane for 5 min at 4 wk of age in two trials. Birds were classified as halothane sensitive (HAL+) or halothane nonresponder (HAL-), in which HAL+ birds showed signs of muscle rigidity in the legs upon removal from halothane gas, and HAL- birds showed no stiffness response. Approximately 3.5% (45) of the turkeys were HAL+. All HAL+ birds and an equal number of HAL- birds were grown until 20 wk of age. Immediately prior to slaughter, all birds were transported in coops on a flatbed trailer for 2 h and then immediately slaughtered upon arrival at the processing plant. Breast muscle pH (0, 1.5, and 24 h postmortem) and L* value (1.5 h and 24 h postmortem) were measured on the fillets. Drip loss and cook loss were also determined on marinated and nonmarinated breast fillets from each carcass. There were no significant mean differences in any parameter measured between the HAL+ and HAL- turkeys. However, the HAL+ turkeys had a greater percentage of fillets with L* values >51 compared with the HAL- turkeys. These results suggest that either halothane response is only a limited predictor of PSE meat in turkeys or that transportation is not an appropriate stressor to induce the PSE condition. PMID- 10824971 TI - Proceedings of the workshop "sperm-mediated gene transfer: advances in sperm cell research and applications." Siena, Italy May 23-26, 1999. PMID- 10824972 TI - CREM: a master-switch regulating the balance between differentiation and apoptosis in male germ cells. AB - Cyclic AMP-responsive element modulator (CREM) is a transcription factor highly expressed in the post-meiotic germ cells of the testis. Its pivotal role is to regulate the expression of several germ cell-specific genes, a crucial function as demonstrated by the severe phenotype of mice whose CREM gene was mutated by homologous recombination. CREM-deficient male animals are sterile and display a ten-fold increase in the apoptosis of germ cells. Recent results have shown that CREM needs a tissue-specific co-activator, ACT (activator of CREM in testis) to elicit its regulatory function in testis. PMID- 10824973 TI - DNA condensation by protamine and arginine-rich peptides: analysis of toroid stability using single DNA molecules. AB - Both somatic cells and sperm have been shown to take up exogenous DNA, but the frequency of its integration is usually low. Scanning probe microscopy studies of sperm chromatin and synthetic DNA-protamine complexes indicate that the coiling of DNA into toroidal subunits, a process initiated in the maturing spermatid to prepare its genome for delivery into the egg, can be mimicked by simply adding protamine to DNA in vitro. The increased resistance of DNA-protamine complexes to nuclease digestion and their structural similarity to native sperm chromatin suggest that the packaging of DNA by protamine might offer a new approach for improving the efficiency of DNA uptake by sperm. Decondensation experiments performed with individual DNA molecules have provided a direct measure of the stability of toroids produced using salmon protamine and smaller arginine-rich peptides. These experiments show that the arginine content of protamine-related sequences can have a dramatic effect on their rate of dissociation from DNA. This technique and the information it provides can be used to identify protamine analogs that can be bound to DNA to increase the efficiency of its uptake by sperm and other cells. PMID- 10824974 TI - Interaction of exogenous DNA with the nuclear matrix of live spermatozoa. AB - Sperm chromatin is a highly organized array of protamines and DNA, with the protamines serving to tightly condense the DNA into a compact, defined structure. We have previously demonstrated that the sperm nucleus is an ordered library of DNA organized into functional zones, such as the nuclear matrix and nuclear annulus. Other laboratories have suggested that mouse spermatozoa can interact with exogenous pSV2CAT plasmid DNA. In this work, we explored this interaction and examined the subcellular localization of the exogenous DNA. We found a repeatable association of exogenous DNA with a specific region of the sperm nuclear matrix. This region of the nucleus correlates with the equatorial segment of the sperm head. This interaction requires only a defined fertilization media, transfection quality DNA, and incubation with spermatozoa. PMID- 10824975 TI - Spermatozoon as a vehicle for HIV-1 and other viruses: a review. PMID- 10824976 TI - Generation of genetically modified mice by spermatozoa transfection in vivo: preliminary results. AB - Mouse vas deferens were injected with a plasmid DNA encoding the GFP (Green Fluorescent Protein). The night after injection males were mated with normal oestrus females, and the offspring were analyzed. From 53 newborns, 4 were found positive by PCR for the GFP gene. In these positive animals, some tissues showed expression for GFP as evidenced by a strong green cytoplasmic fluorescence. GFP expression was particularly patent in the liver (hepatocytes), kidney (renal corpuscle and tubules), abdominal wall, and lung. These preliminary results indicate the possibility to use this method as a simple alternative procedure to create transgenic animals, and it could be especially helpful in species in which the microinjection procedure is not feasible. PMID- 10824977 TI - Nucleosomal domains of mouse spermatozoa chromatin as potential sites for retroposition and foreign DNA integration. AB - Exogenous DNA molecules are spontaneously taken up by sperm cells, internalized in nuclei, and eventually integrated in the sperm genome. The actual occurrence of the integration suggests that the sperm chromosomal DNA is not uniformly and tightly packed with protamines, implying the existence of genomic sites where the chromosomal DNA is accessible to foreign molecules. We have characterized a hypersensitive, nucleosomal subfraction of mouse sperm chromatin that is highly enriched in unmethylated retroposon DNA from a variety of families. Here we propose that both the integration of exogenous DNA molecules, and the endogenous retroposition activity, occur in the same site(s) of sperm chromatin. PMID- 10824978 TI - Intracellular and intercellular transport of many germ cell mRNAs is mediated by the DNA- and RNA-binding protein, testis-brain-RNA-binding protein (TB-RBP). AB - Functions ranging from RNA transport and translational regulation to DNA rearrangement and repair have been proposed for the DNA- and RNA-binding protein, testis-brain-RNA-binding protein (TB-RBP). TB-RBP is primarily in the nuclei of male germ cells during meiosis and in the cytoplasm of male cells after metaphase I of meiosis. Based on its shift in subcellular locations as germ cells differentiate and its binding to microtubules and microfilaments, a model is presented proposing an involvement of TB-RBP in mRNA transport from nucleus to cytoplasm and in the sharing of mRNAs transcribed from the sex chromosomes by movement through intercellular bridges of germ cells. PMID- 10824979 TI - Human spermatogenesis as a model to examine gene potentiation. AB - The first tier of control over the expression of genic domains utilizes chromatin structure. Before the onset of transcription, the chromatin domain that encompasses the gene(s) must assume an open conformation. This renders large segments of the genome available to the tissue-specific and ubiquitous trans factors necessary for proper expression of the genes present. This process has been termed potentiation. It is a necessary obligate, but alone it is not sufficient for gene expression. Spermatogenesis, the development of a viable fertile male gamete, provides a unique model to begin to address the underlying mechanism(s) governing differentiation and tissue-specific gene expression. Male gametogenesis is typified by the activation of numerous genes whose products have novel functions, as well as testis-specific forms of constitutively expressed somatic genes. We have shown that mouse spermatogenesis represents a selective potentiative process (Kramer et al., 1998: Development 125:4749-4655), but little is known about its human counterpart. To fill this void we have examined the potentiative state of several spermatid-expressed genes during the latter stages of human spermatogenesis. We have shown that spermatidexpressed genes are potentiated by the pachytene stage of differentiation. Furthermore, we establish that a chromatin domain functions as a discrete structural unit during differentiation. Interestingly, some of these open structures are maintained in the mature spermatozoon. PMID- 10824980 TI - Analysis and significance of messenger RNA in human ejaculated spermatozoa. AB - Human ejaculate spermatozoa contain multiple mRNA species carried over from earlier stages in spermatogenesis. To date, gene-specific RT-PCR or in situ hybridization has detected transcripts for beta-actin, heat shock proteins (HSP) 70 and 90, protamines (PRM) 1 and 2, transition protein (TNp) 2, HLA II, beta integrins, and, most recently, phosphodiesterase subtypes. We have further evidence for a complex population of transcripts based on screening a human testis cDNA library with a heterologous spermatozoal probe. High levels of transcribed repetitive sequences are present in human spermatozoa, including medium reiteration repeats (MERs) and short and long nuclear interspersed repeats (SINES and LINES). Both SINES and LINES belong to the retroposon class of repeat elements, which are thought to proliferate via an intermediate RNA that is converted to DNA by an RNA-dependent DNA polymerase (reverse transcriptase). We have circumstantial evidence for the presence of an RT in ejaculate sperm based on the detection of transcripts for ORF2 of LINE 1 encoding such an enzyme. Our data suggests the following: 1. Ejaculate spermatozoa may be a very useful tool in the identification of genes linked to an infertile phenotype. 2. Spermatozoa (or spermatids) may express a reverse transcriptase, the role of which is unknown. 3. RNA isolated from spermatozoa or washed semen samples may facilitate the detection of mutations and deletions in testis-expressed AZF-linked genes. PMID- 10824981 TI - Formation of the sea urchin male pronucleus in cell-free extracts. AB - At fertilization, the dormant sperm nucleus undergoes morphological and biochemical transformations leading to the development of a functional male pronucleus. We have investigated the formation of the male pronucleus in a cell free system consisting of permeabilized sea urchin sperm nuclei incubated in fertilized sea urchin egg extract containing membrane vesicles. The first sperm nuclear transformation observed in vitro is the disassembly of the sperm nuclear lamina as a result of lamin B phosphorylation mediated by egg protein kinase C. The conical sperm nucleus then decondenses into a spherical pronucleus in an ATP dependent manner. The new nuclear envelope (NE) forms by ATP-dependent binding of vesicles to chromatin and GTP-dependent fusion of vesicles with one another. Three cytoplasmic vesicle fractions with distinct properties are required for the formation of the male pronuclear envelope. Binding of each fraction to chromatin requires two detergent-resistant lipophilic structures at each pole of the sperm nucleus, which are incorporated into the NE by membrane fusion. Targeting of the bulk of NE vesicles to chromatin is mediated by a lamin B receptor (LBR)-like integral membrane protein. The last step of male pronuclear formation involves nuclear swelling. Nuclear swelling is associated with import of soluble lamin B into the nucleus and growth of the NE. In the nucleus, lamin B associates with LBR, which apparently tethers the NE to the lamina. Thus, formation of the male pronuclear envelope involves a highly ordered series of reactions. PMID- 10824982 TI - Interaction of gametes with exogenous genes: possible opportunities for incorporation into embryonic genome. AB - The mechanism of incorporation of foreign DNA into newly fertilized eggs is poorly understood. It is not known with certainty if S phase DNA replication is required or if integration could occur at other times of the cell cycle that involve DNA strand breaks, such as chromatin rearrangements. We have investigated DNA strand breaks in mouse eggs and zygotes with a sensitive terminal uridine nucleotide end labeling (TUNEL) assay. Greater than 90% of all polar bodies and metaphase II chromosomes in freshly ovulated mouse eggs are TUNEL-assay positive. Approximately one-third of zygotes assayed 6 hr after fertilization contain at least one TUNEL assay positive pro-nucleus and/or decondensing sperm head. These results indicate that early embryonic DNA contains multiple transient DNA breaks that could play a role in the incorporation of foreign DNA. PMID- 10824983 TI - Sperm surface proteins in mammalian fertilization. AB - Boar seminal plasma was separated into five protein fractions (I-V) (> 100, 55, 45, 30, 5-15 kDa) by gel chromatography on Sephadex G-75 SF at pH 7.2. RP-HPLC of protein fractions I-V and N-terminal sequencing of their individual components revealed that the high-molecular-weight aggregates consisted mainly of DQH sperm surface protein and AQN, AWN, PSP II spermadhesins, whereas fraction IV consisted of heterodimers of PSP spermadhesins only. Spermadhesins as monomers were present in seminal plasma in a very low amount. Aggregates containing the DQH protein and AWN spermadhesins as well as HPLC-separated monomeric proteins interacted strongly with acidic polysaccharides. The strongest interaction was observed between biotinylated glycoproteins of porcine zona pellucida and AWN 1-containing aggregates and separated proteins. PSP II interacted with some acidic polysaccharides, whereas the fraction IV corresponding to heterodimer PSP I/PSP II did not show any binding to acidic polysaccharides and zona pellucida. Aggregates containing AWN, AQN, DQH, PSP II proteins, and their separated monomeric forms (fractions I-III) interacted with phosphorylcholine. Fractions I III showed affinity to cholesterol. Biotinylated aggregates containing AWN, AQN, DQH, and PSP proteins (fractions I-IV) bound stronger to boar epididymal spermatozoa than to ejaculated spermatozoa. These results suggest that under physiological conditions, the aggregates of seminal plasma proteins (DQH, AQN, AWN, PSP II) rather than the individual proteins might take part in coating the sperm surface, in sperm capacitation, and in primary binding of spermatozoa to zona pellucida of the ovum. PMID- 10824984 TI - Sperm-mediated gene transfer studies on zebrafish in Singapore. AB - This paper describes the work conducted in the Department of Biological Sciences, National University of Singapore; on sperm-mediated gene transfer in zebrafish. This study began by carrying out direct interaction experiments of pUSVCAT reporter DNA with spermatozoa. Other constructs, including pXGH5, pMTL, pRSVL and pGEM-luc, were subsequently used. The different constructs were taken up by sperm cells with comparable efficiencies. In general, no reporter gene expression, Mendelian inheritance, or evidence of genomic integration of the foreign sequences were obtained. However, transmission of the reporter DNA through generations was observed. DNA uptake by sperm cells was shown using FISH and was enhanced by electroporation. The potential use of more recent approaches, such as REMI and ICSI are explored. Future research directions are discussed. PMID- 10824985 TI - Electroporated sperm mediation of a gene transfer system for finfish and shellfish. AB - We investigated gene transfer in finfish and shellfish via electroporated sperm. The mobility of sperm, the fertilization rate, the hatching rate, gene transfer rate, and abnormality rate of derived embryos were primarily dependent on the voltage level and concentration of DNA during electroporation. Optimal conditions for sperm of each species of aquatic animals can be reached. Genome of the electroporated sperm was analyzed by PCR, and it was shown that an expected-sized product was amplified, corresponding to that of the transgene's amplification. Southern blotting also showed that a positive band located at the same position as the DNA fragment used for the transfer was found in the electroporated sperm after DNase treatment. When the genome isolated from embryos, larvae, juvenile, and adult individuals, all derived from sperm electroporated with foreign DNA molecules, was analyzed by PCR, the existence of foreign DNA was detected in some samples. The integration of the transferred DNA into the genome of transgenic samples was also shown by Southern blot analysis. There was a mosaic distribution of exogenous DNA in a wide variety of tissues analyzed. In addition to CAT activity being positive for the experimental larvae, the transferred GH gene was functional in transgenic finfish and shellfish and resulted in fast-growing transgenic varieties. The overall evidence strongly suggests that the use of electroporated sperm is the simplest yet most efficient approach to perform mass gene transfer in aquacultural animals, including marine mollusks. PMID- 10824986 TI - Electroporation of salmon sperm for gene transfer: efficiency, reliability, and fate of transgene. AB - Uptake of exogenous DNA by electroporated salmon sperm for gene transfer is being investigated. Our studies show that electroporated salmon sperm cells were more efficient and more reliable than untreated sperm in picking up exogenous DNA and subsequently transferring the DNA into salmon embryos. Indirect evidence suggest that some of the exogenous DNA was internalized in the sperm nuclei. The taken up DNA retained its integrity as demonstrated by PCR. The foreign DNA was detected in 15-month-old fish, and had a mosaic pattern of distribution. Integration of the foreign DNA occurred infrequently, and the expression of the foreign genes was poor. The potential of sperm-mediated gene transfer as a routine protocol for mass gene transfer in salmon will be dependent on the improvement of integration and expression of the foreign gene. PMID- 10824987 TI - Sperm/DNA interaction: DNA binding proteins in sperm cell of silkworm Bombyx mori. AB - Plasmid DNA transferred into testes of silkworm larvae following in vivo gene transfer method was taken up by the sperm cells during spermatogenesis and the DNA was transferred to Go and subsequent progeny. RT-PCR analysis confirmed the internalization of the DNA taken up by the sperm. Specific classes of DNA binding proteins present in the sperm act as potential substrates for sperm/DNA interaction and internalization of DNA. Plasmid DNA injected into the testes were rescued from spermatheca, eggs (moths copulated with injected males), and progeny. The rescued plasmid DNA showed CAT activity to the same level as that of control plasmid. PMID- 10824988 TI - DNA interaction with rabbit sperm cells and its transfer into ova in vitro and in vivo. AB - Exogenous DNA interaction with rabbit sperm cells and its transfer into ova has been studied. It was shown that rabbit sperm cells are able to capture, hold, and transfer exogenous DNA to ova during fertilization. Sperm/DNA transfer leads to genetically transformed offspring. Transfection efficiency varied within a large range. Low expression and rearrangements of the transgene were observed. PMID- 10824989 TI - Sperm-mediated preparation of transgenic Xenopus laevis and transmission of transgenic DNA to the next generation. AB - Sperm-mediated transgenesis of frogs Xenopus laevis with retroviral Rous sarcoma virus DNA is described and the transgenic progeny is characterized. Correlation between the high expression of src gene and defective morphogenesis of frog embryos is discussed. PMID- 10824990 TI - DNA dose and sequence dependence in sperm-mediated gene transfer. AB - We have tested three parameters in sperm-mediated gene transfer assays with mice and pigs: (i) the epididymal versus ejaculated origin of sperm cells, (ii) the primary structure, and (iii) the amount of the challenging foreign DNA. We have found that the pVLCNhGH construct, of retrotransposon origin, causes a massive embryo lethality and yet increases the yield of genetic transformation among born animals of both species compared to viral constructs. Arrest of embryonic development is a DNA dose-dependent effect, which is observed with high DNA doses, while lower doses are compatible with development. Finally, the overall efficiency of sperm-mediated gene transfer is higher when ejaculated, versus epididymal, spermatozoa are used. We suggest that this difference is related to the highly efficient apoptotic response in epididymal compared to ejaculated spermatozoa, triggered by the interaction of exogenous DNA molecules with the sperm membrane. PMID- 10824991 TI - Gene integration into bovine sperm genome and its expression in transgenic offspring. AB - Transgenic bovine sperm were produced by restriction enzyme mediated insertion (REMI). REMI utilizes lipofection of linearized pEGFP and the corresponding restriction enzyme for integration into the sperm genomic DNA. The transgenic sperm were used in IVF to produce morula expressing GFP. When transgenic sperm were used for AI in two cows, the resultant calves expressed the exogenous DNA in their lymphocytes as determined by (a) PCR and RT-PCR; (b) specific emission of green fluorescence by GFP; and (c) Southern blot analysis. Data demonstrate that REMI is an efficient method for the production of transgenic sperm and corresponding offspring by AI or embryos by IVF. PMID- 10824992 TI - Modular retro-vectors for transgenic and therapeutic use. AB - Retroviruses have been used for many years as vectors for human gene therapy as well as for making transgenic animals. However, the efficient insertion of genes by retroviruses is often complicated by transcriptional inactivation of the retroviral long terminal repeats (LTRs) and by the production of replication competent retroviruses (RCR). Solutions to these and other difficulties are being found in modular vectors, in which the desirable features of different vector systems are combined. Examples of synergistic vectors include virosomes (liposome/virus delivery), adeno-retro vectors, and MLV/VL30 chimeras. As gene delivery systems become increasingly complex, methodology is also needed for precise assembly of modular vectors. Gene self-assembly (GENSA) technology permits seamless vector construction and simultaneous, multifragment assembly. PMID- 10824993 TI - Sperm-mediated DNA transfer to cells of the uterus and embryo. AB - There is a paucity of information about sperm-mediated transmission of exogenous DNA to implanting embryos and cells of the reproductive tract. Preliminary experiments established that sperm has the capacity to actively take in exogenous DNA derived from HPV. In addition, blastocysts also take up exogenous HPV DNA, but in contrast to sperm, the process appears passive. DNA-carrying sperm migrating in an artificial glass tube or excised mouse bicornuate uteri transfected the blastocysts at the remote position using a flip-flop mechanism. There were preferential transmission of the types of HPV DNA but this was not attributed to the gene sequence or the size of the DNA fragments. The internalized DNA became undetectable unless continuous sperm bombardment or pricking took place. Mycoplasma vectors offer a novel way to enhance the transfection of blastocyst with exogenous DNA. PMID- 10824994 TI - Hijacking oocyte DNA repair machinery in transgenesis? AB - Transgenesis refers to the modification of a genome to carry specified exogenous DNA sequences (transgenes, tgs) in a plant or animal; tgs are ideally transmissible through the germline. It has recently been shown that a membrane disrupted spermatozoon mixed with exogenous DNA can be microinjected into an unfertilized, metaphase II (mII) oocyte to generate transgenic mouse embryos and offspring. This is here referred to as metaphase II (mII) transgenesis. Exogenous DNA clearly becomes genomically integrated in this process, but how? Presumably, the integration process utilizes DNA repair and recombinational machinery resident within mII oocytes. With recent advances in the description of DNA recombination and repair per se and in relation to meiotic and mitotic cell cycles, we are now poised to explain features of mIl transgenesis. Conversely, the method may of itself provide a new tool to probe these aspects of DNA metabolism. This article describes mil transgenesis in the context of DNA recombination and homeostasis in mII oocytes. A fuller understanding of the underlying recombinational mechanisms may enable improved methods of manipulating mammalian genomes and lead to gene targeting and genetic surgery in mII oocytes. PMID- 10824996 TI - Proceedings of the workshop "sperm-mediated gene transfer: advances in sperm cell research and applications." Siena, Italy May 23-6, 1999. Conclusions PMID- 10824995 TI - TransgenICSI reviewed: foreign DNA transmission by intracytoplasmic sperm injection in rhesus monkey. AB - This brief review considers the status of transgenesis by intracytoplasmic sperm injection (ICSI) with nonhuman primates. GFP expressing rhesus macaques embryos (mean = 34.6%; N = 81) were produced by ICSI using rhodamine-tagged DNA encoding the green fluorescence protein (GFP) gene bound on sperm. Rhodamine signal was lost at the egg surface during in vitro fertilization (IVF) but could be traced by dynamic imaging during ICSI within the egg cytoplasm. GFP gene was expressed as early as the 4-cell stage in ICSI embryos but not in embryos produced by in vitro fertilization (IVF). The percentage of GFP expressing blastomeres increased during embryogenesis to the blastocyst stage. Three offspring resulted from seven embryo transfers-a set of anatomically normal twins (a male and a female) stillborn 35 days premature, and a healthy male born at term. Although transgene was not detected in the offspring, the successful production of live primates using DNA bound sperm by ICSI suggests an alternative route to creating transgenic animals. It also raises concern regarding transmission of infectious material during ICSI. PMID- 10824997 TI - A t(2;19)(p13;p13.2) in a giant invasive cardiac lipoma from a patient with multiple lipomatosis. AB - Cardiac lipomas occur infrequently but account for a significant portion of rare cardiac tumors. Common cutaneous lipomas have previously been associated with rearrangements of chromosome band 12q15, which often disrupt the high-mobility group protein gene HMGIC. In this report, we describe the cytogenetic analysis of an unusual giant cardiac lipoma that exhibited myocardial invasion in a patient with a history of multiple lipomatosis (cutaneous lipoma, lipomatous gynecomastia, lipomatous hypertrophy of the interatrial septum, and dyslipidemia). Cytogenetic studies of cells derived from the cardiac lipoma demonstrated no abnormalities of chromosome 12, but did reveal a t(2;19)(p13;p13.2). A liposarcoma-derived oncogene (p115-RhoGEF) previously mapped to chromosome 19 and the low-density lipoprotein receptor gene (LDLR) previously mapped to chromosome band 19p13 were evaluated to determine whether they were disrupted by this translocation. Fluorescence in situ hybridization analyses assigned p115-RhoGEF to chromosome 19 in bands q13.2-q13.3 and mapped the LDLR to chromosome arm 19p in segment 13.2, but centromeric to the t(2;19) breakpoint. Thus, these genes are unlikely to be involved in the t(2;19)(p13;p13.2). Further studies of the regions of chromosomes 2 and 19 perturbed by the translocation in this unusual infiltrating cardiac lipoma will identify gene(s) that participate in adipocyte growth and differentiation and may provide insight into syndromes of multiple lipomatosis. PMID- 10824998 TI - MOZ is fused to p300 in an acute monocytic leukemia with t(8;22). AB - We report on the fusion of the monocytic leukemia zinc finger protein (MOZ) gene to the adenoviral E1A-associated protein p300 (p300) gene in acute monocytic leukemia M5 associated with a t(8;22)(p11;q13) translocation. We studied two patients with double-color fluorescence in situ hybridization (FISH) using the yeast artificial chromosome 176C9 and the bacterial artificial chromosome clone H59D10 specific to the MOZ and p300 genes, respectively. Both probes were split in the patients' chromosome metaphase cells, and the two derivative chromosomes were each labeled with both probes. We showed by Southern blot the rearrangement of the MOZ gene, and cloned the fusion transcripts in one patient carrying the t(8;22) by reverse transcription-polymerase chain reaction using MOZ- and p300 specific primers. Both fusion transcripts were expressed. This result defines a novel reciprocal translocation involving two acetyltransferases, MOZ and p300, resulting in an abnormal transcriptional co-activator that could play a critical role in leukemogenesis. PMID- 10824999 TI - Adrenocortical carcinoma is characterized by a high frequency of chromosomal gains and high-level amplifications. AB - Distinction of adrenocortical carcinoma from benign adrenocortical lesions by standard criteria is often difficult. In order to search for additional diagnostic parameters, a series of 25 adrenocortical tumors, 8 adenomas, 14 primary carcinomas, 1 metastasis, and the 2 adrenocortical carcinoma cell lines SW13 and NCI-H295 were analyzed by the approach of comparative genomic hybridization (CGH). Except for the two smallest adenomas, all tumors showed chromosomal imbalances with a high incidence of chromosomal gains, most frequently involving chromosomes or chromosome arms 5, 7, 8, 9q, 11q, 12q, 14q, 16, 17q, 19, 20, and 22q. The only significant loss of material concerned the distal part of 9p. Furthermore, 21 high-level amplifications were identified in 15 different regions of the genome. The consensus regions of recurrent gains and the focal high-level amplifications allowed identification of a series of chromosomal subregions containing candidate proto-oncogenes of potential pathogenic function in adrenocortical tumors: 1p34.3-pter, 1q22-q25, 3p24-pter, 3q29, 7p11.2-p14, 9q34, 11q12-11q13, 12q13, 12q24.3, 13q34, 14q11.2-q12, 14q32, 16p, 17q24-q25, 19p13.3, 19q13.4, and 22q11.2-q12. A subset of the CGH data was independently confirmed by interphase cytogenetics. Interestingly, the adenomas larger than 4 cm contained gained material of regions also overrepresented in carcinomas. In addition, several chromosomal gains, in particular the high-level amplifications, were exclusive for the malignant status of the tumors. These data indicate that the larger adrenal lesions need to be carefully considered in the diagnosis of adrenocortical tumors, and that genetic aberrations might provide useful markers for a better diagnostic differentiation. PMID- 10825000 TI - A consistent pattern of RIN1 rearrangements in oral squamous cell carcinoma cell lines supports a breakage-fusion-bridge cycle model for 11q13 amplification. AB - Gene amplification is a common feature of tumors. Overexpression of some amplified genes plays a role in tumor progression. Gene amplification can occur either extrachromosomally as double-minute chromosomes (dmin) or intrachromosomally in the form of homogeneously staining regions (hsrs). Approximately one-half of our oral squamous cell carcinomas (OSCCs) are characterized by amplification of band 11q13, usually as an hsr located entopically (occurring or situated at the normal chromosomal site, as opposed to ectopically). Using chromosomal fluorescence in situ hybridization (FISH), we confirmed the amplification of the cyclin D1 (CCND1/PRAD1) and fibroblast growth factor types 3 and 4 (FGF3/INT2 and FGF4/HSTF1) genes within the 11q13 amplicon in our series of primary OSCCs and derived cell lines. The human RIN1 gene was isolated as an RAS interaction/interference protein in a genetic selection in yeast and has been described as a putative effector of both the RAS and ABL oncogenes. We mapped RIN1 to 11q13.2. FISH analysis of 10 11q13-amplified OSCC cell lines revealed high-level RIN1 amplification in two cell lines. Three additional cell lines have what appear to be duplications and/or low-level amplification of RIN1, visible in both interphase and metaphase cells. The hybridization pattern of RIN1 on the metaphase chromosomes is particularly revealing; RIN1 signals flank the 11q13 hsr, possibly as a result of an inverted duplication. The gene amplification model of Coquelle et al. (1997) predicted that gene amplification occurs by breakage-fusion-bridge (BFB) cycles involving fragile sites. Our data suggest that the pattern of gene amplification at 11q13 in OSCC cell lines is consistent with a BFB model. RIN1 appears to be a valuable probe for investigating the process of gene amplification in general and, specifically, 11q13 amplification in oral cancer. PMID- 10825001 TI - Susceptibility gene for familial acute myeloid leukemia associated with loss of 5q and/or 7q is not localized on the commonly deleted portion of 5q. AB - The molecular mechanism for the occurrence of leukemia in multiple members of a family has not been fully elucidated but data support the contribution of highly penetrant mutations in leukemia susceptibility genes. We have investigated the genetic etiology of an unusual three-generation family with apparent autosomal dominant transmission of acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS) accompanied by somatic loss of the long arm of chromosome 5 and/or loss of heterozygosity (LOH) analysis and fluorescence in situ hybridization (FISH) of leukemia cells have been performed, confirming acquired hemi- and homozygous deletion of the long arm of chromosome 5. However, the chromosome lost in the observed LOH event is from the affected parent, in contradiction to the expectation for a two-hit hypothesis involving a tumor suppressor gene. Furthermore, genetic linkage has been performed at 5q31-33 as well as other loci (21q22 and 16q21-23.2) previously implicated in familial leukemia. In this family, linkage analysis excludes loci at 5q31-33 and 21q22, but localization to 16q21-23.2 cannot be excluded. We observed a maximum multipoint LOD score of 1.19 between marker D16S265 and D16S503 at 16q22 (P = 0.03), suggesting possible linkage to this locus. Considering this family and the previous 16q-linked family together, the linkage of a leukemia susceptibility gene to 16q22 achieved an LOD score of 3.63 at D16S265 with theta = 0. Thus, somatic deletion of the long arm of chromosome 5 appears as a necessary but surprisingly noncausative event for onset of AML and MDS in this family, thereby confirming a multistep etiology in which chromosome 5 plays an important secondary role. PMID- 10825002 TI - Allelotype analysis of flow-sorted breast cancer cells demonstrates genetically related diploid and aneuploid subpopulations in primary tumors and lymph node metastases. AB - Flow cytometric DNA content measurements have demonstrated extensive DNA ploidy heterogeneity in primary breast carcinomas. However, little is known at the molecular level about the clonal relationship between these tumor cell subpopulations, or about the molecular genetic changes associated with aneuploidization. We have used flow cytometric cell sorting to dissect some of this complexity by isolating clonal subpopulations in breast carcinomas for comparative molecular genetic analysis. Clonal subpopulations were isolated from 12 primary breast carcinomas and 5 lymph node metastases from 4 cases based on DNA content and cytokeratin 8/18 labeling. DNA from these clones was screened for allelic imbalances with 92 polymorphic microsatellite markers mapped to 39 different chromosome arms. Diploid and aneuploid populations were concurrently present in 11 out of 12 primary tumors. The DNA ploidy status of primary tumors was identical to that of the related lymph node metastases. Allelic imbalance was present in 10 out of 11 diploid clones (mean, 3.4 +/- 4.2). All allelic imbalances observed in the diploid clones recurred in the cognate aneuploid clones, but were, in the latter, accompanied by additional allelic imbalances at other loci and/or chromosome arms (mean, 10.9 +/- 5.8). In only two of the four metastatic cases did the allelotypes of metastatic clones show small differences relative to their cognate primary tumors. The primary diploid tumor clone recurred in all lymph node metastases. This study indicates that the majority of allelic imbalances in breast carcinomas are established during generation of DNA ploidy diversity. Recurrence of the allelic imbalances in diploid clones in the aneuploid clones suggests linear tumor progression, whereas the simultaneous presence of early diploid and advanced aneuploid clones in both primary and metastatic tumor sites suggests that acquisition of metastatic propensity can be an early event in the genetic progression of breast cancer. PMID- 10825003 TI - Genomewide assessment of genetic alterations in DMBA-induced rat sarcomas: cytogenetic, CGH, and allelotype analyses reveal recurrent DNA copy number changes in rat chromosomes 1, 2, 4, and 7. AB - Rat sarcomas, induced by subcutaneous injections of 7,12 dimethylbenz[a]anthracene (DMBA), were studied with the objective of identifying critical chromosome regions associated with tumorigenesis. We employed three genomewide screening techniques-cytogenetics, CGH, and allelotyping-in 19 DMBA induced sarcomas in F1 (BN/Han x LE/Mol) rats. The most conspicuous finding in the cytogenetic analysis was a high incidence of trisomy for rat chromosome 2 (RNO2). Signs of gene amplification (hsr) were also seen in several tumors. The CGH analysis revealed that gains in copy number were much more common than losses. The gains mostly affected RNO2 (10/19), RNO12q (7/19), and RNO19q (5/19), as well as the proximal part of RNO4 (8/19) and the distal part of RNO7 (7/19). Reduction in copy number was seen in RNO17 (2/19). For the allelotyping, we used 318 polymorphic microsatellite marker loci covering the entire genome. We identified regions of allelic imbalance affecting most of the rat chromosomes. The highest incidences of recurrent allelic imbalance were observed at loci in certain regions in RNO1, 2, 4, and 7 and at lower incidences in parts of RNO12, 16, 18, and 19. The combined results suggested that genetic alterations detected in RNO2 and RNO12 usually corresponded to complete or partial trisomy, whereas those in RNO1 and RNO7 seemed to involve regional deletions and/or gains. Furthermore, we could confirm that copy number gains occur proximally in RNO4, where a previous study showed amplification of the Met oncogene in a subset of these tumors. PMID- 10825004 TI - Lack of correlation between expression of retinoic acid receptor-beta and loss of heterozygosity on chromosome band 3p24 in esophageal cancer. AB - Loss of heterozygosity (LOH) on chromosome arm 3p occurs frequently in human cancers, including esophageal cancer, suggesting that tumor suppressor genes may be located on this chromosome arm. The retinoic acid receptor-beta (RARB) gene is localized on chromosome band 3p24, and its expression is progressively lost during esophageal carcinogenesis. Furthermore, growth inhibition of esophageal cancer cell lines by all-trans retinoic acid has been associated with the constitutive and induced expression of RARB. We therefore assessed LOH on chromosome arm 3p and RARB expression in esophageal cancer to investigate the mechanism of altered RARB expression during carcinogenesis. We first analyzed LOH in 65 paired surgical specimens of normal mucosa and esophageal cancer by using 10 microsatellite markers, which resulted in 44 informative cases for subsequent study. LOH on chromosome band 3p24 was found to occur at an overall rate of 36.4% (16/44) by three markers (D3S1293, THRB, and D3S1283). LOH for these three individual markers was 14.0%, 47.4%, and 20.9%, respectively. Meanwhile, RARB expression was lost in 43.2% (19/44) of these 44 samples. The loss of RARB expression was not correlated with LOH on chromosome band 3p24 (gamma = -0.22, 0.069, and -0.02, P = 0.15, 0.78, and 0.9 for D3S1293, THRB, and D3S1283, respectively), although both altered RARB expression and LOH in esophageal cancer were statistically significant (P = 0.001 and 0.0001, respectively), indicating that the loss of RARB expression cannot be explained by LOH on 3p24. PMID- 10825005 TI - Mapping the amplification of EIF3S3 in breast and prostate cancer. AB - Gain of chromosome arm 8q is a frequent genetic alteration in breast and prostate cancer. Two amplified subregions, 8q21 and 8q23-24, have been identified with comparative genomic hybridization (CGH). We have recently demonstrated that the EIF3S3 (eIF3-p40) gene, located at 8q23, is often amplified and overexpressed in both breast and prostate cancer. Here, we used fluorescence in situ hybridization (FISH) to map the amplified region around EIF3S3 in primary breast cancers and cell lines. The size of the common highly amplified region was about 2.5 Mb between the markers D8S1668 and WI-7959. Next, we analyzed the expression of all expressed sequence tags (ESTs) located within and near this region by RNA slot blot hybridization. In addition to EIF3S3, three anonymous ESTs and EXT1 were found to be highly expressed in cancer cell lines with the amplification at 8q23 q24. However, the anonymous ESTs were located outside the minimal highly amplified region and EXT1 was overexpressed only in one of the cancer cell lines with 8q amplification. Since EIF3S3 was the only consistently overexpressed gene located in the minimal highly amplified region, it is the strongest candidate target gene for 8q23-q24 amplification. PMID- 10825006 TI - High frequency of clonal chromosome abnormalities in prostatic neoplasms sampled by prostatectomy or ultrasound-guided needle biopsy. AB - Cancer of the prostate remains poorly characterized cytogenetically. This is due in part to methodological problems and in part to the paucity of radical prostatectomies, until now the main source of material for cytogenetic analyses. We have improved existing techniques for the culturing of prostatic neoplasms removed by radical prostatectomy or sampled by ultrasound-guided needle biopsy. Successful short-term cultures were obtained from all 10 prostatectomy samples and from all 10 ultrasound-guided needle biopsies, always with a pure epithelial morphology. Of the 19 cases yielding a sufficient number of high-quality metaphases for chromosome banding analysis, the single atypical epithelial hyperplasia had a normal karyotype, whereas both prostatic intraepithelial neoplasias and 12 of 16 (75%) invasive carcinomas were shown to have clonal abnormalities. Ten of the 12 (83%) karyotypically abnormal invasive carcinomas presented structural chromosomal rearrangements. A recurrent deletion, del(10)(p13), was seen in three tumors; in one of them the terminal nature of the deletion was confirmed by two-color FISH. A del(17)(p11) was seen in one PIN lesion, but since the analysis of exons 4-8 of the TP53 tumor suppressor gene revealed no mutations, there probably was no inactivation of the second TP53 allele. Our study thus leads to the following main conclusions. First, better culturing methods allow the detection of abnormal karyotypes in a much higher percentage of prostatic neoplasms than has hitherto been possible. Second, ultrasound-guided needle biopsies of prostatic neoplasms are a sufficient source of material for cytogenetic analysis. Third, a terminal deletion of the short arm of chromosome 10, del(10)(p13), seems to identify a subgroup of prostatic cancer. PMID- 10825007 TI - A novel and consistent amplicon at 13q31 associated with alveolar rhabdomyosarcoma. AB - Rhabdomyosarcomas are the most common soft-tissue sarcoma found in children. The alveolar subtype is clinically more aggressive than the embryonal subtype. In addition to the presence of specific chromosome translocations and associated fusion gene products in a high proportion of the alveolar subtype, we previously showed that tumors with this histology frequently show evidence of genomic amplification. Here, we substantially extended the number of alveolar rhabdomyosarcoma samples examined by comparative genomic hybridization analysis. Regions of loss were noted, including the smallest overlapping regions corresponding to 16q, 17/17p, and 9q32-34, in 16%, 10%, and 10% of cases, respectively (44 primary samples/6 cell lines). Amplification or gain at 12q13-15 in the region of the MDM2/GLI1/SAS/CDK4 loci and 2p24 at the MYCN locus was found in 28% and 32% of cases, respectively. Single amplicons were found at locations that in other samples showed consistent gain, including the regions 5q15-23, 7q21 31, 11p11-14, 17q23-24, and 20q13, and amplification was found in two cases at 15q24-26. However, most striking was a novel region of amplification or gain at 13q31 in 19% of cases (51 primary samples/6 cell lines). This indicates that a gene or genes at 13q31 are significant in the development or progression of alveolar rhabdomyosarcoma. PMID- 10825008 TI - Concurrent translocations of MLL and CBFA2 (AML1) genes with new partner breakpoints in a child with secondary myelodysplastic syndrome after treatment of acute lymphoblastic leukemia. AB - The MLL gene at 11q23 is frequently disrupted by chromosomal translocations in de novo acute myeloid leukemia (AML) and acute lymphoid leukemia (ALL), and in secondary leukemia induced by treatment with inhibitors of topoisomerase II, including the epipodophylotoxins. The CBFA2 gene at 21q22 is also frequently disrupted in de novo ALL and AML and less commonly in secondary AML. Rearrangements of MLL and CBFA2 have been described in de novo and secondary myelodysplastic syndrome (MDS). There have been no previous descriptions of coexisting abnormalities of MLL and CBFA2 in cases of MDS or acute leukemia. We describe a patient who developed secondary MDS after chemotherapy for hyperdiploid ALL. At the time of conversion to MDS, the patient had 46 chromosomes, with an 11q23/MLL translocation involving a new partner breakpoint at 2p23 and a 21q22/CBFA2 translocation involving a new partner breakpoint at 6p22. This report is the first to describe new partner breakpoints at 2p23 and 6p22 for MLL and CBFA2 genes, respectively, and concurrent rearrangements of these genes in a patient with secondary MDS. PMID- 10825009 TI - Variant translocations involving 16q22 and 17p13 in solid variant and extraosseous forms of aneurysmal bone cyst. PMID- 10825010 TI - 2000 Directory of mass spectrometry manufacturers and suppliers. PMID- 10825011 TI - Liquid-liquid extraction using 96-well plate format in conjunction with liquid chromatography/tandem mass spectrometry for quantitative determination of methylphenidate (Ritalin) in human plasma. AB - Methylphenidate (MPH; Ritalin: methyl-alpha-phenyl-2-piperidinacetate hydrochloride) is utilized for the treatment of attention deficit hyperactivity disorder (ADHD) and narcolepsy. Recently, we described a rapid enantioselective liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for the determination of the enantiomers of MPH (Rapid Commun. Mass Spectrom. 1999; 13: 2054). A lower limit of quantification (LLOQ) of 87 pg/mL was attained for the human plasma assay. The present paper describes a high-throughput sample preparation procedure in conjunction with racemic LC/MS/MS analysis for MPH with a LLOQ of 50 pg/mL. A semi-automated robotics method using liquid-liquid extraction (LLE) in a 96-well plate format was developed and validated. The correlation coefficients were > or =0.998 for MPH indicating good fits of the regression models over the range of the calibration curve. The accuracy and precision of the semi-automated approach were comparable to those obtained using the manual sample preparation technique reported previously (vide supra). The current method can easily be adapted to the enantioselective LC/MS/MS assay of MPH. The assay was simple, fast, specific, and exhibited excellent ruggedness. PMID- 10825012 TI - Formation of a2+ ions of protonated peptides. An ab initio study. AB - The mechanism of the formation of a2+ ions from b2+ ions occurring during fragmentation of protonated peptides is investigated using quantum chemical methods. The geometries of the stationary structures involved in two possible mechanisms, namely, a two-step mechanism via an open-chain acylium ion and a concerted pathway involving rupture of two covalent bonds of the cyclic isomer of the b2+ ion, as well as the energetics of the reactions, were calculated at the MP2 and B3LYP levels, both combined with the 6-31G(d,p) as well as the 6 31++G(d,p) basis sets for the simplest analog of the b2+ ion. The energetically favored path is the direct expulsion of the CO molecule from the cyclic b2+ ion. The ZPE-corrected barrier height for this reaction is 26.2 kcal mol(-1) at the MP2/6-31G(d,p) level, while the highest barrier along the two step path is 31.4 kcal mol(-1). The barrier height for the reverse reaction is 3.8 kcal mol(-1), significantly smaller than the average kinetic energy release (KER) measured for larger b2+ ions. The barrier height for the reverse reactions of the MeCO-NH CHMeCO+, NH2-iBuCH-CO-NH-CH2CO+, and NH2-CH2-CO-NH-CH(i-Bu)CO+ b2+ ions was found to be 11.3, 9.6, and 18.4 kcal mol(-1), in reasonable agreement with the measured KER for these reactions, indicating that the simplest model compound has unique properties in this respect. Based on comparisons with G2-MP2 calculations, comments are made on the applicability of various levels of theory for the description of the reaction. PMID- 10825013 TI - Detection of oxidative species for 4-phenoxyphenol derivatives during the electrospray ionization process. AB - Analyses by flow injection as well as liquid chromatography/mass spectrometry (LC/MS) and liquid chromatography/tandem mass spectrometry (LC/MS/MS) were performed with four 4-phenoxyphenol derivatives. When ambient temperature nitrogen gas was used to facilitate solvent evaporation, [M + H]+, [M + NH4]+, and [2M + NH4]+ ions were observed as the major ions. As the nitrogen gas temperature increased from ambient to 250 and 450 degrees C, [M]+*, [M - 1]+ and [M + 15]+ ions were the predominant ions. Heat-induced oxidation was found to be the primary source for the formation of oxidative species. Aqueous solvents were found to be essential for the formation of the [M + 15]+ ions. The [M]+* and [M + 15]+ ions were further characterized by tandem mass spectrometry. Based on the MS/MS data, it was proposed that the [M + 15]+ ions were the in-source generated 1,2-quinone ions. PMID- 10825014 TI - Mass spectrometric study of six cyclic esters AB - A series of cyclic esters, which are optically active as a consequence of their helical structures, were synthesized to investigate the relationships between their structures and their optical activities. This paper reports the electron impact fragmentation mechanisms of these six cyclic esters. Accurate mass measurements and mass analyzed ion kinetic energy spectrometry confirmed fragmentation patterns. The stability of the fragment ions has a great influence on the fragmentation pathways, but no correlation with the optical activity was found. PMID- 10825015 TI - Stopped-flow electrospray ionization mass spectrometry: a new method for studying chemical reaction kinetics in solution. AB - In this work a new mass spectrometry based method for monitoring the kinetics of chemical reactions in solution is described. A stopped-flow mixing instrument is coupled to an electrospray ionization (ESI) mass spectrometer via a novel type of interface. Chemical reactions are initiated by rapid mixing of two reactant solutions. The mixture is instantaneously transferred to a reaction tube where the kinetics can be monitored in real-time by ESI mass spectrometry. With the current setup, a time window from 2.5 to 36 seconds after mixing of the reactants can be monitored. The experimental setup is used to study the kinetics of acetylcholine hydrolysis under alkaline conditions as a function of pH. The intensities of reactant (acetylcholine) and product (choline) ions are monitored simultaneously as a function of time. The reaction is carried out under pseudo first-order conditions and the intensity-time curves are well described by single exponentials. The rate constants determined from these fits compare favorably with previous data from the literature. PMID- 10825016 TI - Generation of hydrogen radicals for reactivity studies in Fourier transform ion cyclotron resonance mass spectrometry. AB - An efficient technique for generation of H* (D*) radicals in Fourier transform ion cyclotron resonance (FTICR) mass spectrometry is described. The method allows the probing of the reactivity of gas-phase H* radicals towards various ions isolated in the cell of an FTICR mass spectrometer. Results on interactions of H* and D* radicals with trapped positive or negative C60 fullerene ions, as well as singly charged peptide ions, are presented. Hydrogen radical addition or H/D exchange reactions between trapped ions and free H* (D*) radicals were observed. Potential implementation of the technique for probing the gas-phase three dimensional structures of polyatomic ions is discussed as well. PMID- 10825017 TI - Single bead and hard tag decoding using accurate isotopic difference target analysis-encoded combinatorial libraries. AB - A single fully automated generic microbore liquid chromatography electrospray time-of-flight protocol has been developed for the analysis of single beads, single beads with analytical constructs, and isotopically labelled dialkylamine hard tags. The protocol relies upon the incorporation of an isotopic signature into the linker and/or derivatising molecule to give a specific isotopic ratio together with an accurate isotopic difference. These properties facilitate highly specific and sensitive analysis of beads and hard tags using accurate isotopic difference analysis (AIDA) without prior knowledge of the molecular weight. Three open access methods with automated processing have been developed around a core generic approach. PMID- 10825018 TI - Mechanism of cross-ring cleavage reactions in dirhamnosyl lipids: effect of the alkali ion. AB - Liquid secondary ion mass spectrometry and high-energy collision-induced dissociation were used to analyze a dirhamnosyl lipid mixture. The negative fast atom bombardment spectrum reveals a mixture of four homologous dirhamnosyl lipids with the following general structure: Rha-Rha-Cn-Cm (where Cn and Cm denote 3 hydroxy fatty acid moieties). The mass region 450-600 u in the collision-induced dissociation spectra of the negative [M - H]- ions shows product ions that can be rationalized by terminal loss of a 3-hydroxyalkanoic acid residue; these ions can be used for the characterization of the fatty acid substituents. A unique effect of alkali-metal ions on the course of fragmentation of dirhamnosyl lipid attachment ions was observed. The strong chelation of sodium is revealed from the stability of the [M - H + 2Na]+ ion that does not lose a sodium ion with the eliminated neutrals, contrary to what is observed for the dilithium adduct. Cross ring cleavages occur during high-energy collision-induced dissociation of both positively and negatively charged precursor ions. The results suggest a concerted decomposition pathway involving the six-membered rings of the monosaccharide residues. The formation of cross-ring cleavage products, which retain the C10-C10 moiety during high-energy collision-induced dissociation of all the precursor ions that contain sodium or lithium, strongly supports a retro [2 + 2 + 2] mechanism. PMID- 10825019 TI - Ni-Zr alloys: relationship between surface characteristics and electrocatalytic behavior AB - A relationship between electrocatalytic activity for the hydrogen evolution reaction and the surface composition of the electrode was established for Ni-Zr crystalline and amorphous alloys by means of secondary ion mass spectrometry (SIMS). Electrocatalytic activity was tested by means of cathodic polarization in 1 M KOH at 25 degrees C and the resulting exchange current density has been taken as a measure of catalytic efficiency. Surface activation treatment involved chemical etching in HF solutions; the consequent morphological and compositional surface changes were studied by differential scanning calorimetry (DSC), X-ray diffraction (XRD) and scanning electron microscopy (SEM). The electrochemical behavior of the pure elements (Ni and Zr) was also considered for comparison. All samples submitted to chemical etching in HF solutions showed an increase in electrocatalytic activity, particularly the alloy with the highest Ni content. The beneficial effect of chemical etching is due to dissolution of the zirconium oxide layer and to the formation of nanocrystalline Ni on the surfaces. PMID- 10825020 TI - Hydrogen/deuterium exchange using a coaxial sheath-flow interface for capillary electrophoresis/mass spectrometry. AB - The interfacing of capillary electrophoresis (CE) with mass spectrometry (MS) is well established and may be accomplished by use of either a coaxial arrangement or by employing a liquid T-junction. In both these interfaces a make-up flow is introduced. This is required because of the mismatch in flow rates for capillary electrophoresis approximately nL/min and 'true' electrospray approximately 2-10 microL/min. Electrical connectivity may also be established where the liquid flows meet (the introduction of nanospray renders the use of make-up flow unnecessary). Hydrogen/deuterium (H/D) exchange occurs in solution when there are labile hydrogen atoms present in a molecule. The establishment of the presence and the number of such exchangeable hydrogen atoms may be of importance in the identification and differentiation of compounds. It may also be an aid in the structural elucidation of unknown materials. We have investigated the feasibility of carrying out H/D exchange via a CE/MS interface. This involved the addition of D2O to the sheath flow and our preliminary results showing the separations of drug substances, subsequently undergoing exchange, are presented. PMID- 10825021 TI - Diarrhea and AIDS in the era of highly active antiretroviral therapy. PMID- 10825022 TI - Introduction: integrating geriatrics into the subspecialty of infectious diseases. PMID- 10825023 TI - A perspective on cellular immunity in the elderly. AB - The increased incidence of infection and malignancy in elderly individuals has prompted many studies that demonstrate that the aging immune system is impaired. Most of these studies have focused on the impairment of acquired immunity provided by lymphocytes. While defects in acquired humoral and T-cell-mediated immunity may exist, increased susceptibility to infection may result from defects in the constitutive functioning of macrophages and granulocytes. Recognition of the potential importance of defects in constitutive immunity in the elderly may provide new opportunities for therapeutic and prophylactic intervention in this population. PMID- 10825024 TI - Antigen presentation in the immune response to infectious diseases. AB - T lymphocytes mediate immune defenses against infectious diseases. The major histocompatibility complex class I and class II antigen-processing pathways play an essential role in the activation of pathogen-specific T lymphocytes by presenting peptide fragments derived from pathogen-encoded proteins. The past 5 years have seen remarkable advances in our understanding of both of these antigen processing pathways. Many new proteins that are critical to T-cell-mediated defenses against infectious pathogens have been identified and characterized. Although the direct effects of aging on antigen processing and presentation are relatively unexplored, it is likely that immunosenescence affects and is affected by the antigen-processing pathways. PMID- 10825025 TI - Micronutrient supplementation and immune function in the elderly. AB - Immunologic function, particularly cell-mediated immunity, declines with age, contributing to the increased incidence of infectious diseases in the elderly. Nutrition may play a pivotal role in maintaining immune competence in older adults. Most studies to date have focused on micronutrient deficiencies and supplementation, sometimes using "mega-dose" formulations. Multivitamin/mineral supplements or specific micronutrients such as zinc and vitamin E may be of value; however, data suggest there is likely a therapeutic range for many micronutrients, and oversupplementation may be harmful. Specific alterations of dietary lipids may also be useful for modulating immune responses in the elderly. This review summarizes the prevalence of vitamin and mineral deficiencies in older adults and highlights the outcomes of trials of micronutrient supplementation to augment immune function in the elderly. PMID- 10825027 TI - Pneumonia in the elderly: overview of diagnostic and therapeutic approaches. AB - Pneumonia is more frequent in the elderly and results in higher morbidity and mortality. Although the incidence of pneumonia increases with age, from 1 per 1,000 to 12 per 1,000 persons over age 75 years, comorbid medical illnesses and host defense impairments (especially heart disease, chronic obstructive pulmonary disease, and aspiration risk) are independent risk factors. The microbial etiology of pneumonia in the healthy elderly is similar to that in younger patients but shifts toward a more gram-negative and opportunistic flora with increasing age and severity of concomitant medical illness. The choice of antimicrobial therapy must be based on risk stratification (age, medical illnesses, and severity of presentation). Guidelines based on these principles will be reviewed. Pneumococcal and influenza vaccination reduce the risk of death due to pneumonia and are cost-effective preventative strategies. PMID- 10825026 TI - Factors contributing to susceptibility of postmenopausal women to recurrent urinary tract infections. AB - Recurrent urinary tract infections (UTIs) account for substantial morbidity, losses in work time, and medical costs. Recent studies suggest that the major factors predisposing to recurrent UTIs differ by age and functional status. In premenopausal women, sexual intercourse, spermicide exposure, maternal history of UTI, and a history of UTI in childhood have been associated with recurrent infections. In postmenopausal women, lack of estrogen appears to be an important factor predisposing to recurrent UTI, as does nonsecretor status, a history of UTI in the premenopausal period, incontinence, presence of a cystocele, and postvoid residual urine. In older women who are institutionalized, catheterization, incontinence, antimicrobial exposure, and functional status are most strongly related to risk of recurrent UTI. Further research is needed to better identify measures for prevention of recurrent UTI, which likely differ by age group. PMID- 10825028 TI - Pneumococcal drug resistance: the new "special enemy of old age". AB - Streptococcus pneumoniae is a leading cause of illness and death among the elderly. The recent emergence of drug-resistant strains has complicated selection of antimicrobial therapy for suspected pneumococcal infections. In some areas of North America, nearly 40% of pneumococcal isolates from the blood or cerebrospinal fluid of persons > or = 65 years old had reduced susceptibility to penicillin. Of all penicillin-resistant infections, >30% occur in persons > or = 65 years old. The increasing prevalence of drug-resistant pneumococci and recent outbreaks of pneumococcal disease in chronic-care facilities emphasize the importance of efforts to prevent these infections in the elderly. Limiting selection for drug-resistant strains through judicious use of antimicrobial drugs and preventing invasive pneumococcal infections through increased use of pneumococcal vaccine form the foundation of these efforts. PMID- 10825029 TI - Herpes zoster in the elderly: issues related to geriatrics. AB - This article reviews specific clinical and research issues of herpes zoster related to geriatric medicine. Salient epidemiological and clinical issues include the increasing probability of zoster and postherpetic neuralgia with aging, age-related decline in immunity to varicella-zoster virus, the functional and psychosocial impact of zoster on the quality of life of the elderly, illness behavior in elderly patients with zoster, and varicella-zoster virus transmission and control in the nursing home. The role of antiviral therapy, corticosteroids, and analgesics; the measurement and analysis of pain, health-related quality of life, and functional status; and development of the varicella vaccine in the management of zoster in the elderly are also emphasized. Fertile research opportunities exist within these areas for investigators interested in infectious diseases, geriatrics, and other zoster-related disciplines. PMID- 10825030 TI - AIDS and the elderly. AB - Individuals 50 years of age or older continue to account for at least 10% of AIDS cases reported to the Centers for Disease Control and Prevention in recent years. Little research is devoted to addressing the specific issues affecting diagnosis, treatment, and prevention of AIDS in older Americans. Survival rates among elderly individuals infected with human immunodeficiency virus (HIV) are consistently decreased in comparison with those for younger patients. Elderly individuals also are less likely to use a condom during sexual intercourse or to participate in routine HIV testing. This article reviews the current literature concerning the changing epidemiology of AIDS among older Americans. The article also addresses AIDS-related morbidity and mortality, treatment issues, and HIV prevention behaviors among the elderly. Enhanced clinician awareness of HIV in the elderly, along with further research concerning HIV treatment and prevention, is necessary to improve survival and outcome for those patients. PMID- 10825031 TI - Atherosclerosis and infection due to Chlamydia pneumoniae or cytomegalovirus: weighing the evidence. AB - A link between infectious agents and atherosclerosis has been postulated for decades. This review describes the epidemiological and biological evidence linking cytomegalovirus and Chlamydia pneumoniae to atherosclerotic disease. Case control studies and histologic evidence from atheromatous specimens support an association between atherosclerosis and infection with these two microorganisms, and small interventional trials appear to confirm the link with C. pneumoniae, but these findings require confirmation in larger studies. A lack of clinically relevant animal models has hampered investigations regarding biological mechanisms, particularly for C. pneumoniae. PMID- 10825032 TI - Antibiotic use in the elderly: issues and nonissues. AB - Despite the well-recognized increase in mortality and morbidity due to infections in the elderly, antibiotics may, in most cases, be used in a manner similar to that in younger patients. The decreased lean body weight and reduced renal function typical of elderly patients, however, require consideration of reduced doses and longer dosing intervals, especially for renally excreted antibiotics. Length of therapy should be conservative because underlying anatomic or functional predispositions to infections tend to complicate treatment. Oral antibiotics are equally well absorbed in the elderly and younger patients and may be used for the same indications as for younger patients. A notable, important difference in the choice of antibiotics for serious infections in older versus younger patients is that empirical therapy should be broader in spectrum for elderly patients, and especially for elderly long-term residents, since the variety of infecting bacteria tends to be greater and polymicrobial infections tend to be common. PMID- 10825033 TI - Integrating geriatrics into clinical training, research training, board certification, and continuing education in infectious diseases: meeting review and commentary. AB - Although adults aged 65 years and older constitute the most rapidly growing segment of the U.S. population, geriatric issues have not typically been a focus of training in infectious diseases (ID). Underrecognition of the unique aspects of geriatric care, apathy toward this population, and the feeling that "we're all geriatricians" (and thus know geriatric medicine) all contribute to this problem. This article summarizes the recent meeting focused on integrating geriatric principles within ID training at all levels. The ID/geriatric interface as an attractive area for basic and clinical research is emphasized. PMID- 10825034 TI - Pulmonary actinomycosis. PMID- 10825036 TI - Diagnosis of pneumococcal pneumonia in epidemiological studies: evaluation in Kenyan adults of a serotype-specific urine latex agglutination assay. AB - A serotype-specific tube latex agglutination assay for 10 pneumococcal serotypes was evaluated with use of urine samples from 72 Kenyan adults with pneumonia whose blood or lung aspirate cultures were positive for Streptococcus pneumoniae, 203 patients with pneumonia whose cultures were negative for S. pneumoniae, and 101 afebrile controls. Detection thresholds for purified capsular polysaccharide in normal urine ranged from 0.33 to 10 ng/mL. The sensitivity of the assay for the 10 pneumococcal serotypes was 0.57 (95% confidence interval, 0.44-0.70) and was unaffected by human immunodeficiency virus seropositivity, prior antibiotic use, and bacteremic or nonbacteremic status but varied significantly by serotype. Of the pneumococci obtained by culture, 81% were of serotypes (1, 4, 5, 6, 7, 9, 12, 14, 19, and 22) that were included in the antigen assay. Strong simultaneous agglutinations against two different serotypes were found in urine samples from two patients. The specificity of the assay was 0.98 (lower 95% confidence limit, 0.95). Subjective reading of agglutination results introduced variation in specificity that may be inapparent if not formally measured. The assay extended the diagnostic yield in pneumococcal pneumonia by a factor of 2.2 (from 54 diagnoses established by blood culture to 119 established by both methods) and may therefore prove useful in reducing the sample size of epidemiological studies of pneumococcal pneumonia in adults. PMID- 10825035 TI - Usefulness of the cytomegalovirus (CMV) antigenemia assay for predicting the occurrence of CMV disease and death in patients with AIDS. AB - A cohort study of 214 human immunodeficiency virus (HIV)-infected patients was performed to assess the usefulness of the cytomegalovirus (CMV) antigenemia assay for predicting the occurrence of CMV disease and death. Multivariate analysis revealed that only positive baseline CMV antigenemia assays (relative risk [RR], 7.2; 95% confidence interval [CI], 3.7-14.2; P = .0001) and CD4 cell counts (RR, 0.98; 95% CI, 0.97-0.99; P = .009) were associated with CMV disease. A positive baseline CMV antigenemia assay was also associated with death by multivariate analysis (RR, 2.2; 95% CI, 1.5-3.4; P = .0003). Increasing levels of CMV antigenemia during follow-up were associated with increased risks of CMV disease and death. A positive CMV antigenemia assay that showed > 10 cells per 2 x 10(5) polymorphonuclear leukocytes during follow-up was 91% sensitive and 84% specific for predicting a diagnosis of CMV disease; the negative predictive value for this positive test was high (97%). Therefore, the CMV antigenemia assay appears to be a simple, rapid, and inexpensive test for predicting the occurrence of CMV disease and death in patients with advanced HIV infection. PMID- 10825037 TI - Randomized, placebo-controlled trial of HA-1A, a human monoclonal antibody to endotoxin, in children with meningococcal septic shock. European Pediatric Meningococcal Septic Shock Trial Study Group. AB - Meningococcal septic shock has a rapid onset and characteristic skin hemorrhages that allow bedside diagnosis. Initial plasma endotoxin levels are high and correlate closely with clinical outcome. In a double-blind, randomized, placebo controlled trial (planned, n = 270; actual, n = 269), we compared the effectiveness of HA-1A (6 mg/kg of body weight iv; maximum, 100 mg), a human monoclonal antibody to endotoxin, and placebo in reducing the 28-day all-cause mortality rate among children with a presumptive clinical diagnosis of meningococcal septic shock. Treatment groups were well balanced for baseline characteristics and prespecified prognostic variables. In this trial no significant benefit of HA-1A could be demonstrated. The 28-day mortality rates in the intention-to-treat analysis were as follows: placebo, 28%; HA-1A, 18%; reduction in mortality, 33% (P = .11, per Fisher's exact test, two-tailed; odds ratio = 0.59; 95% confidence interval for the difference, 0.31-1.05). All patients tolerated HA-1A well, and no antibodies to HA-1A were detected. PMID- 10825038 TI - Hepatosplenic cat-scratch disease in children: selected clinical features and treatment. AB - We reviewed 19 cases of hepatosplenic cat-scratch disease at Texas Children's Hospital (Houston). The range of the patients' ages was 2 years 4 months to 11 years 8 months. The chief complaint was fever for all patients. The duration of fever before diagnosis was 7 to 56 days (mean, 22 days). Abdominal pain was present in 13 patients (68%). Thirteen children were treated with rifampin alone, and three received rifampin therapy plus gentamicin or trimethoprim sulfamethoxazole. Once rifampin therapy was initiated alone or in combination, improvement was noted within 1 to 5 days (mean, 2.6 days) for patients who had had prolonged fever the duration of which before rifampin therapy averaged 3 weeks. The most common dosage and duration for our patients were 20 mg/[kg x d] every 12 hours and 14 days, respectively. Rifampin should be considered in the initial antimicrobial treatment of hepatosplenic cat-scratch disease. PMID- 10825039 TI - Clinical reactogenicity of intradermal bacille Calmette-Guerin vaccination. AB - Clinical, microbiological, and immunologic responses were evaluated in volunteers vaccinated intradermally with bacille Calmette-Guerin (BCG). Most volunteers (98%) developed ulcerative lesions that drained for a mean +/- SE of 4.3 +/- 0.29 weeks. Mycobacterial DNA was detected by a polymerase chain reaction-based amplification technique in biopsy specimens from BCG ulcers 2 weeks after vaccination and in blood specimens 3 days after vaccination. Mycobacteria were cultured from ulcer drainage 2 months after vaccination, demonstrating a prolonged potential risk of contact spread of the vaccine strain. The duration of ulcer drainage was inversely correlated with prevaccination lymphoproliferative (r = -0.515; P < .002) and interferon gamma (r = -0.841; P < .002) responses specific to mycobacteria and directly correlated with postvaccination increases in lymphoproliferative (r = 0.498; P < .002) and interferon gamma (r = 0.688; P < .02) responses specific to mycobacteria. These results demonstrate the clinical reactogenicity of BCG and the potential risk of contact spread of the vaccine strain and suggest that clinical reactogenicity is a trade-off for the induction of protective mycobacterial immunity. PMID- 10825040 TI - Editorial response: variation in clinical and immune responses to bacille Calmette-Guerin--implications for an improved tuberculosis vaccine. PMID- 10825041 TI - The role of red blood cell polymorphisms in resistance and susceptibility to malaria. AB - In regions highly endemic for Plasmodium falciparum malaria, red cell polymorphisms that confer resistance to severe disease are widespread. Sickle cell trait, alpha-thalassemia, glucose-6-phosphate dehydrogenase deficiency, and blood groups were determined in 100 children from Gabon with severe malaria who were matched with 100 children with mild malaria and followed up for evaluation of reinfections. The sickle cell trait was significantly associated with mild malaria and blood group A with severe malaria. During follow-up, the original severe cases had significantly higher rates of reinfection than the original mild cases, with higher parasitemia and lower hematocrit values. Incidence rates did not differ in the context of erythrocyte polymorphisms, but patients with sickle cell trait presented with markedly lower levels of parasitemia than those without. Thus, the severity of malaria is partly determined by the presence of blood group A and the sickle cell trait. The different presentation of reinfections in severe versus mild cases probably reflects different susceptibility to malaria. PMID- 10825042 TI - Intravenous immunoglobulin therapy for streptococcal toxic shock syndrome--a comparative observational study. The Canadian Streptococcal Study Group. AB - Twenty-one consecutive patients with streptococcal toxic shock syndrome (TSS) between December 1994 and April 1995 were treated with a median dose of 2 g of intravenous immunoglobulin (IVIG)/kg (cases) and were compared with 32 patients with streptococcal TSS between 1992 and 1995 who did not receive IVIG therapy (controls). The outcome measure was 30-day survival. Patient plasma was tested for its ability to inhibit T cell activation induced by the infecting strain. The proportion of cases with 30-day survival was higher than that of the controls with 30-day survival (67% vs. 34%, respectively; P = .02). Multivariate analysis revealed that IVIG administration and a lower Acute Physiology and Chronic Health Evaluation II score were associated with survival; the odds ratio for survival associated with IVIG therapy was 8.1 (95% confidence interval, 1.6-45; P = .009). IVIG therapy enhanced the ability of patient plasma to neutralize bacterial mitogenicity and reduced T cell production of interleukin-6 and tumor necrosis factor alpha. IVIG may be an effective adjunctive therapy for streptococcal TSS, possibly because of its ability to neutralize bacterial exotoxins. PMID- 10825044 TI - Perinatal transmission of hepatitis G virus (GB virus type C) and hepatitis C virus infections--a comparison. AB - Hepatitis G virus (HGV) infection is more common than hepatitis C virus (HCV) infection and is frequently found in healthy individuals. Although parenteral spread of HGV is well recognized, other routes of transmission probably occur as well. In a prospective study of mother-to-infant transmission of hepatitis viruses, 69 pregnant women with antibodies to HCV and their 81 newborn children were included. Serum levels of HCV RNA and HGV RNA were detected by polymerase chain reaction (PCR) assays, and antibodies to HCV and HGV envelope protein E2 were detected by enzyme-linked immunosorbent assay. Fifty-nine of the mothers had HCV viremia, whereas 16 had HGV viremia. HCV transmission from viremic mothers occurred in 2.8%-4.2% of the cases, whereas HGV transmission from viremic mothers occurred in 75.0%-80.0% of the cases (P < .001). Sequencing of the PCR products of HGV from the mother-infant serum pairs showed minor differences in most cases but sequence homology in each pair. Although the rate of perinatal HGV transmission highly exceeded that of perinatal HCV transmission, HGV did not seem to induce hepatitis in the children. PMID- 10825043 TI - The effects of mefloquine treatment in pregnancy. AB - We investigated the relationship between mefloquine antimalarial treatment and the outcome of pregnancy in Karen women living in an area along the western border of Thailand where multidrug-resistant Plasmodium falciparum infections are common. Of 3,587 pregnancies investigated, 208 (5.8%) were exposed to mefloquine, 656 (18.3%) to quinine only, and 909 (25.3%) to other antimalarials, and 2,470 (68.9%) had no documented malaria. There were 61 stillbirths and 313 abortions. Women who received mefloquine treatment during but not before pregnancy had a significantly greater risk of stillbirth than did women treated with quinine alone (odds ratio [OR], 4.72; 95% confidence interval [CI], 1.7-12.7), women exposed to other treatments (OR, 5.10; 95% CI, 2-13.1), and women who had no malaria (OR, 3.50; 95% CI, 1.6-7.6) (P < .01). This association remained after adjustment for all identified confounding factors. Mefloquine was not associated with abortion, low birth weight, neurological retardation, or congenital malformations. Mefloquine treatment during pregnancy was associated with an increased risk of stillbirth. PMID- 10825045 TI - Nontyphoid Salmonella bacteremia: age-related differences in clinical presentation, bacteriology, and outcome. AB - In a retrospective study, 80 episodes of nontyphoid salmonella (NTS) bacteremia in children were compared with 55 episodes in adults over a 10-year period. The study disclosed major differences in the predisposition, clinical presentation, and outcome as well as the microbiology of NTS bacteremia in relation to age. Adults were more likely than children to have predisposing diseases (95% vs. 15%, respectively; P < .0001) and to receive prior medications (95% vs. 23%, respectively; P < .0001), particularly immunosuppressive agents (58% vs. 5%, respectively; P < .0001). In most adults (67%), NTS infection presented as a primary bacteremia and was associated with a high incidence of extraintestinal organ involvement (34%) and a high mortality rate (33%). In children, NTS bacteremia was usually secondary to gastroenteritis (75%) and caused no fatalities. Although group D Salmonella (78%) and the serovar Salmonella enteritidis were the predominant isolates from adults, the emergence of infections due to group C Salmonella (46%) and the serovar Salmonella virchow in children was noted. PMID- 10825046 TI - Nosocomial infections in adult patients undergoing extracorporeal membrane oxygenation. AB - The use of extracorporeal membrane oxygenation (ECMO) for adult patients has increased in recent years. A retrospective cohort study of adult patients undergoing ECMO was performed between 19 February 1985 and 10 October 1995 to evaluate nosocomial infections. Seventy-one evaluable patients underwent ECMO for a total of 799 days. Forty-six infections were identified in 32 (45%) of 71 patients. There were 15 bloodstream infections, 13 lower respiratory infections, 11 urinary tract infections, and 7 miscellaneous infections. The rates of bloodstream infection (18.8 cases per 1,000 ECMO days) and urinary tract infection (13.8 cases per 1,000 ECMO days) were significantly higher than those reported through the Centers for Disease Control and Prevention / National Nosocomial Infection Surveillance System (P < .0001 and P < .001, respectively). The rate of bloodstream infection increased with the duration of ECMO cannulation. This study highlights the increased risk for nosocomial infections in this patient population. Infection may not be apparent, and increased physician awareness of infection risk is imperative. PMID- 10825047 TI - A case-control study after a hantavirus infection outbreak in the south of Belgium: who is at risk? AB - Puumala is the most common hantavirus serotype in Europe and is spread mainly by the red bank vole. Between 1 July 1992 and 31 January 1994, an outbreak of Puumala virus-induced nephropathia epidemica (NE) occurred in the Belgian Ardennes. Serologically confirmed cases (n = 41) were compared with two groups of asymptomatic seronegative controls. Risks identified included sighting of living rodents, exposure to rodent droppings, and trapping rodents during the 4 weeks preceding onset of symptoms. Activities during this 4-week period that presented the greatest risk were woodcutting, reopening of a nonaerated room, and strenuous physical effort. This is the first case-control study on risk factors for NE in Europe. In comparison with the American form of hantavirus pulmonary syndrome, which is spread by deer mice, professional activity appears to be a more important risk factor for acquisition of hantavirus in Europe. PMID- 10825048 TI - Failure of physicians to consider the diagnosis of pertussis in children. AB - To determine the ability of physicians to make a diagnosis of pertussis and factors associated with improved diagnosis, 8,235 children from 88 child care centers and 14 elementary schools from Quebec City, Quebec, Canada, were evaluated by using a questionnaire completed by parents and a medical record review. Children must have consulted a physician to be included in the evaluation. There were 558 children meeting the surveillance case definition and 416 meeting a modified World Health Organization case definition who consulted a physician. A diagnosis of pertussis was considered in 24%-26% of children meeting either case definition, made in 12%-14%, and reported for 6%. Pertussis diagnosis was significantly associated with having a history of pertussis exposure (P < or = .003), four pertussis-related symptoms (P < .001), and a cough for > or = 5 weeks (P < or = .05) and consulting in a hospital setting (P < or = .03). The proportion of cases of pertussis diagnosed and reported is low even when children present with classical symptoms. PMID- 10825049 TI - Determination of failure of treatment of plasmodium falciparum infection by using polymerase chain reaction single-strand conformational polymorphism fingerprinting. AB - The inability to distinguish failures of treatment of Plasmodium falciparum infection from new infections is an important impediment to the evaluation of antimalarial drugs. On the basis of a pilot study utilizing polymerase chain reaction (PCR) single-strand conformational polymorphism (SSCP) analysis to genotype P. falciparum isolates, we sought to confirm that PCR SSCP analysis could reliably distinguish infections for which treatment failed from unrelated infections with a sample size adequate to estimate the accuracy of this technique. PCR SSCP analysis of the MSP-1, MSP-2, and GLURP genes was performed on 72 paired isolates recovered from 36 individuals for whom treatment failed in Thailand. In every case (100% [95% confidence interval (CI), 90%-100%]), the PCR SSCP pattern of the recrudescent isolates matched that of the primary isolate. We determined whether PCR SSCP analysis could separate unrelated infections by comparing each recrudescent isolate with each of the unrelated primary isolates. Of 1,260 comparisons, 1,258 (99.8% [95% CI, 99.4%-100%]) were unique. The results indicate that PCR SSCP analysis can be used to differentiate infections for which treatment failed from reinfections. PMID- 10825050 TI - Family cluster of Rocky Mountain spotted fever. AB - Soon after a patient from Tennessee died of Rocky Mountain spotted fever (RMSF), several family members developed symptoms suggestive of the disease and were treated presumptively for RMSF. Fifty-four persons visiting the index patient's home were interviewed; serum samples were collected from 35. Three additional cases of RMSF were confirmed, all of which occurred in first-degree relatives. Time spent at the family home and going into the surrounding woods were significantly associated with developing antibodies to Rickettsia rickettsii. Ticks were collected and examined for rickettsiae by polymerase chain reaction analysis. Because hyperendemic foci and family clusters of RMSF can occur, when a case is suspected clinicians should be vigilant for signs and symptoms consistent with R. rickettsii infection in other persons who may have been similarly exposed. PMID- 10825051 TI - Hemorrhagic fever with renal syndrome in the Dolenjska region of Slovenia--a 10 year survey. AB - This report describes the first investigation of clinical findings for a larger series of patients with hemorrhagic fever with renal syndrome (HFRS) who were infected with Dobrava virus. From 1985 to 1995, 38 patients with serologically confirmed HFRS were hospitalized at the regional hospital in Novo mesto in the Dolenjska region of Slovenia. On the basis of results of serological examination, 24 patients had Dobrava virus infection, and 14 patients had Puumala virus infection. Complete clinical data were available for 31 patients. Eleven patients underwent hemodialysis for treatment of acute oliguric or anuric renal failure. Four patients, all infected by Dobrava virus, had signs of shock and severe bleeding. Three severely ill Dobrava virus-infected patients died of hemorrhagic complications. We have demonstrated that Dobrava and Puumala viruses coexist in a single region of endemicity and are capable of causing HFRS with significant differences in severity. PMID- 10825052 TI - Consecutive epidemics of Q fever in a residential facility for drug abusers: impact on persons with human immunodeficiency virus infection. AB - Two large outbreaks of Q fever occurred in 1987 and 1988 in an agricultural community for the rehabilitation of drug users. Approximately 40% of the residents were human immunodeficiency virus (HIV)-positive. Two hundred thirty five residents presented with clinical evidence of a flulike syndrome that was confirmed to be Q fever; moreover, a large proportion of residents developed an asymptomatic infection. Clinical signs and symptoms were rather nonspecific: fever, malaise, and muscle pain that were often associated with pulmonary symptoms. Single or multiple opacities were detected, with mild interstitial inflammation evident on chest roentgenograms. The source of infection was the sheepfold, which is part of the stock-farming activity of the community. Both outbreaks occurred just after lambing had begun. Residents who were exposed during the first epidemic were protected in the second one. The attack rate among HIV-positive residents was significantly higher than that among HIV-negative residents in the first outbreak, whereas only a slight, marginally significant difference was observed in the second outbreak. The clinical features of Q fever did not differ between HIV-positive and HIV-negative individuals. No cases of relapse or chronic disease were observed. PMID- 10825054 TI - Tick-borne encephalitis. AB - Tick-borne encephalitis (TBE) is a zoonotic arbovirus infection endemic to Russia and Eastern and Central Europe. Despite being a common and serious life threatening disease for which a mass vaccination program was implemented in Austria, there is only limited reference to this disease in the English-language literature. TBE is transmitted to humans usually by the bite of a tick (either Ixodes persulcatus or Ixodes ricinus); occasionally, cases occur following consumption of infected unpasteurized milk. Transmission is seasonal and occurs in spring and summer, particularly in rural areas favored by the vector. TBE is a serious cause of acute central nervous system disease, which may result in death or long-term neurological sequelae. Effective vaccines are available in a few countries. The risk for travelers of acquiring TBE is increasing with the recent rise in tourism to areas of endemicity during spring and summer. PMID- 10825053 TI - Streptococcus pneumoniae spinal infection in Nottingham, United Kingdom: not a rare event. AB - Pneumonia and meningitis are the most frequent manifestations of Streptococcus pneumoniae infection. Spinal infection is considered to be a rarity. Between 1985 and 1997, 8 patients with spinal infection (vertebral osteomyelitis, 3; spinal epidural abscess, 1; both, 4) due to S. pneumoniae were seen at University Hospital (Nottingham, U.K.). Predisposing factors for pneumococcal infection were documented for five patients and included diabetes mellitus, alcoholism, and corticosteroid therapy. One patient presented with concomitant meningitis and endocarditis. Clinical features of note were prolonged symptoms and a lack of febrile response. S. pneumoniae was isolated from the blood of five patients. Magnetic resonance imaging was used to localize the spinal infection in five patients. Two cases were managed medically. Three patients died after a protracted illness. A literature search revealed 20 other cases of spinal infections due to S. pneumoniae. The salient features of the cases are summarized. PMID- 10825055 TI - Thalidomide as therapy for human immunodeficiency virus-related oral ulcers: a double-blind placebo-controlled clinical trial. AB - A double-blind, randomized, placebo-controlled clinical trial was performed in Mexico City to evaluate the efficacy of thalidomide in treating oral recurrent aphthae in human immunodeficiency virus (HIV)-infected subjects. Sixteen HIV infected patients with clinical and histological diagnosis of oral recurrent aphthous ulcerations received randomly an 8-week course of either thalidomide or placebo, with an initial oral dosage of 400 mg/d for 1 week, followed by 200 mg/d for 7 weeks. Ten subjects received thalidomide and six received placebo. At 8 weeks, nine subjects (90%) in the thalidomide group had complete healing of their ulcers, compared with two (33.3%) of the six patients in the placebo group (P = .03). There was a significant reduction in largest ulcer diameter in the thalidomide group. Rash was observed in 80% of the thalidomide patients. Although thalidomide demonstrated an unquestionable benefit in treatment of oral ulcers in HIV patients, caution must be taken given the frequent occurrence of side effects. PMID- 10825056 TI - Isoniazid toxicity in health care workers. AB - The toxicity of isoniazid chemoprophylaxis was assessed in 83 health care workers (HCWs) receiving a 6-month course, in whom clinical toxicity and liver function were monitored. Thirty-four HCWs (41%) developed an adverse event; in 26 (76%), toxicity was sufficiently severe to require cessation of treatment. Of the total, liver function test abnormalities (serum alanine transaminase levels more than two times normal) were evident in 14 subjects, with 8 requiring cessation of therapy. Other symptoms reported included malaise, nausea with associated anorexia, arthralgia, and rash. Mean time to development of symptoms was 3 weeks (range, 0.5-6 weeks), with the mean age of those with toxicity not differing significantly from those without (38 vs. 39 years). The high rate of toxicity seen in this study is sufficiently notable that we advocate the use of monthly liver function testing and frequent review in those receiving isoniazid prophylactic therapy. PMID- 10825057 TI - Influenza B virus encephalitis. AB - Acute encephalitis and postinfectious encephalopathy have been reported infrequently in association with influenza A and B virus infections. We report herein a case of a 6-year-old girl with acute influenza B virus encephalitis resulting in neurological sequelae. The diagnosis was made by isolation of influenza B virus from the nasopharynx, seroconversion to influenza B, and reverse transcription polymerase chain reaction (RT-PCR) identification of the virus from the patient's cerebrospinal fluid. Direct sequencing of viral RNA from the patient's nasopharynx and cerebrospinal fluid revealed identical nucleotide sequences in the HA1 region of the hemagglutinin gene. This is the first report of influenza B virus encephalitis diagnosed by use of RT-PCR and illustrates the need for increased awareness of influenza virus as a cause of acute encephalitis. PCR may be a useful tool for diagnosing future cases. PMID- 10825058 TI - Effect of a 14-day course of foscarnet on cytomegalovirus (CMV) blood markers in a randomized study of human immunodeficiency virus-infected patients with persistent CMV viremia. Agence National de Recherche du SIDA 023 Study Group. AB - A randomized open-label phase 2 trial compared the virological and clinical effects on cytomegalovirus (CMV) infection of a 14-day course of intravenous foscarnet (100 mg/[kg x 12 h]) or no treatment in 42 HIV-infected patients with < 100 CD4 cells/mm3 and persistent asymptomatic CMV viremia. All CMV markers (blood culture, pp65 antigenemia, plasma and leukocyte DNA) either became negative or decreased significantly at day 14 in the foscarnet group. CMV blood culture results at day 14 were positive in 14% of those receiving foscarnet versus 60% of control patients (P = .004). However, after the end of treatment, all markers reappeared or the virus load rapidly increased. The probability of CMV disease at 6 months was 43% in both groups. Patients who had or who achieved a negative blood culture at any time had a reduced risk of CMV disease (RR = 2.64; 95% CI = 1.24-5.62; P = .02). This study suggests that sequential courses of intravenous foscarnet might not be a good strategy for preemptive therapy in this population and that in patients with a positive blood marker, treatment able to induce and maintain negative CMV blood cultures could constitute an effective intervention. PMID- 10825059 TI - Increased cell-free viral DNA in fatal cases of chronic active Epstein-Barr virus infection. AB - We have studied the nature of Epstein-Barr virus (EBV) infection in 33 patients with chronic active EBV infection. The study population included 14 patients with fatal chronic EBV infection and 19 patients with nonfatal chronic EBV infection, as well as 18 patients with acute EBV-induced infectious mononucleosis and 10 healthy controls. EBV DNA was measured in serum or plasma samples from the patients by semiquantitative polymerase chain reaction-based assay. EBV DNA was detected in serum or plasma samples from 62% (9/14) of patients with fatal chronic active EBV infection. In contrast, only 11% (2/19) of patients with nonfatal chronic active EBV infection and 11% (2/18) of patients with infectious mononucleosis displayed EBV DNA. None of the healthy controls tested positive. Cell-free circulating EBV DNA may represent an important feature of chronic active EBV infection and may provide a useful tool to monitor the severity of this illness. PMID- 10825060 TI - Primary lamivudine resistance in acute/early human immunodeficiency virus infection. PMID- 10825061 TI - Massive hemoperitoneum: a new manifestation of bacillary peliosis in human immunodeficiency virus infection. PMID- 10825062 TI - Persistent fever as the only manifestation of chronic coxsackievirus B4 infection in the brain of a human immunodeficiency virus type 1-infected child. PMID- 10825063 TI - Postexposure rabies vaccination in patients infected with human immunodeficiency virus. PMID- 10825064 TI - Successful treatment of multiple cerebral histoplasmomas with itraconazole. PMID- 10825066 TI - Adult pneumococcal cellulitis: case report and review. PMID- 10825065 TI - Successful treatment of ventriculitis due to carbapenem-resistant Acinetobacter baumannii with intraventricular colistin sulfomethate sodium. PMID- 10825067 TI - Recrudescence of cutaneous Mycobacterium haemophilum lesions following tetanus immunization. PMID- 10825068 TI - Severe neutropenia during therapy for concurrent primary human immunodeficiency virus and cytomegalovirus infections. PMID- 10825069 TI - Melioidosis brain and lung abscess after travel to Sri Lanka. PMID- 10825070 TI - Candida (Torulopsis) glabrata: a new pathogen found in an empyema. PMID- 10825071 TI - Gas gangrene in an immunocompromised girl due to a Clostridium ramosum infection. PMID- 10825072 TI - Disseminated papulopustular eruption due to Mycobacterium fortuitum in an immunocompetent patient. PMID- 10825073 TI - Torovirus gastroenteritis presenting as acute abdomen. PMID- 10825074 TI - Autoimmune thrombocytopenia associated with Borrelia burgdorferi. PMID- 10825075 TI - Arabinose-positive Burkholderia pseudomallei infection in humans: case report. PMID- 10825076 TI - Aerobic and anaerobic microbiology of mycotic aortic aneurysm. PMID- 10825077 TI - Fungemia due to Saccharomyces species in a patient treated with enteral Saccharomyces boulardii. PMID- 10825078 TI - Foscarnet activity on human immunodeficiency virus type 1 in the central nervous system. PMID- 10825079 TI - Hepatitis B vaccination in persons with isolated hepatitis B core antibodies. PMID- 10825080 TI - Native valve infective endocarditis in elderly and younger adult patients: comparison of clinical features and outcomes with use of the Duke criteria. PMID- 10825081 TI - Amphotericin B colloidal dispersion versus amphotericin B in the empirical treatment of fever and neutropenia. PMID- 10825082 TI - Clostridium difficile-associated diarrhea in human immunodeficiency virus infection--a changing scenario. PMID- 10825083 TI - Metronidazole resistance in Helicobacter pylori. PMID- 10825084 TI - Use of spoligotyping in molecular epidemiology of tuberculosis. PMID- 10825085 TI - Salmonellosis associated with chicks and ducklings--Michigan and Missouri, Spring 1999. AB - During the spring of 1999, outbreaks of salmonellosis associated with handling chicks and ducklings occurred in Michigan and Missouri. This report summarizes the epidemiologic information for the outbreaks and provides an overview of legislative efforts to control the distribution of chicks and ducklings. These outbreaks demonstrate that handling chicks and ducklings is a health risk, especially for children, and highlight the need for thorough handwashing after contact with chicks, ducklings, and other young fowl. PMID- 10825086 TI - Measles outbreak--Netherlands, April 1999-January 2000. AB - On June 21, 1999, a cluster of five cases of measles was reported among the 390 students attending a religion-affiliated elementary school in The Netherlands. Persons belonging to this religious denomination routinely do not accept vaccination. Municipal health services (MHSs) investigated and found 160 suspected measles cases among children attending the school. By February 4, 2000, 2961 measles cases, including three measles-related deaths, had been reported by 35 MHSs to the national registry. This report summarizes the investigation of the measles outbreak in The Netherlands, which indicated that measles can be a severe disease among unvaccinated populations in The Netherlands. PMID- 10825087 TI - Fatal yellow fever in a traveler returning from Venezuela, 1999. AB - On September 28, 1999, a previously healthy 48-year-old man from California sought care at a local emergency department (ED) and was hospitalized with a 2 day history of fever (102 F [38.9 C]), chills, headache, photophobia, diffuse myalgias, joint pains, nausea, vomiting, constipation, upper abdominal discomfort, and general weakness. On September 26, he had returned from a 10-day trip to Venezuela. On September 29, an infectious disease physician from the ED contacted the Marin County Health Department (MCHD) about the patient's symptoms; MCHD reported his illness to the California Department of Health Services (CDHS) as a suspected case of viral hemorrhagic fever. This report describes the investigation of the case. PMID- 10825088 TI - "The time has come...." (reflections on leading change) PMID- 10825089 TI - Fathoming the mysteries of magnetic resonance imaging. PMID- 10825090 TI - Clinical findings and coronary artery disease in dogs and cats with acute and subacute myocardial necrosis: 28 cases. AB - Records of final diagnoses based on necropsies performed on dogs and cats over a 4.5-year period at a university teaching hospital were examined for the diagnosis of acute or subacute myocardial necrosis. Clinical findings signaling the occurrence of myocardial necrosis were often not specific, due to simultaneously occurring disease processes. However, of 28 animals identified, dyspnea occurred frequently (17/28; 61%) and in some cases in the presence of minimal pulmonary pathology (2/3; 66%) or otherwise unexplained pulmonary edema (4/4; 100%). Elevations in serum aspartate aminotransferase (10/10; 100%) and creatine kinase (5/9; 55%) were also frequent. Disease processes associated with thrombus formation were present for each case in which a coronary artery thrombus occurred (5/28; 18%). PMID- 10825091 TI - Tubular colonic duplication in a dog. AB - A diagnosis of tubular colonic duplication was made via contrast radiography and colonoscopy in a six-month-old, intact female Boston terrier. Clinical signs of increased frequency of defecation, tenesmus, and constipation, which had been present since birth, resolved following surgical correction of the duplication. The literature addressing diagnosis and treatment of tubular colonic duplication in dogs is reviewed. PMID- 10825092 TI - Effects of methimazole on renal function in cats with hyperthyroidism. AB - The purpose of this study was to investigate the effects of methimazole on renal function in cats with hyperthyroidism. Twelve cats with naturally occurring hyperthyroidism and 10 clinically normal (i.e., control) cats were included in this study. All cats initially were evaluated with a history, physical examination, complete blood count, serum biochemistry profile, basal serum total thyroxine concentration, complete urinalysis, and urine bacterial culture. Glomerular filtration rate (GFR) was estimated by a plasma iohexol clearance (PIC) test. After initial evaluation, hyperthyroid cats were treated with methimazole until euthyroidism was achieved. Both groups of cats were then reevaluated by repeating the initial tests four to six weeks later. The mean (+/ standard deviation) pretreatment estimated GFR for the hyperthyroid cats was significantly higher (3.83+/-1.82 ml/kg per min) than that of the control cats (1.83+/-0.56 ml/kg per min). Control of the hyperthyroidism resulted in a significantly decreased mean GFR of 2.02+/-0.81 ml/kg per minute when compared to pretreatment values. In the hyperthyroid group, the mean increases in serum urea nitrogen (SUN) and creatinine concentrations and the mean decrease in the urine specific gravity after treatment were not statistically significant when compared to pretreatment values. Two of the 12 hyperthyroid cats developed abnormally high serum creatinine concentrations following treatment. These results provide evidence that cats with hyperthyroidism have increased GFR compared to normal cats, and that treatment of feline hyperthyroidism with methimazole results in decreased GFR. PMID- 10825093 TI - Ultrasonographic measurement of gastrointestinal wall thickness and the ultrasonographic appearance of the ileocolic region in healthy cats. AB - Ultrasound evaluation was performed on 11 healthy cats to determine wall thickness measurements for the stomach, duodenum, jejunum, ileum, and colon and to characterize the appearance of the ileocolic region. The terminal ileum had a characteristic "wagon wheel" appearance on cross-sectional images. Gastrointestinal wall thickness measurements were as follows: gastric fundus (mean, 2.0 mm; 95% confidence interval [CI], 1.7 to 2.2 mm), pylorus (mean, 2.1 mm; 95% CI, 1.9 to 2.4 mm), duodenum (mean, 2.2 mm; 95% CI, 2.0 to 2.4 mm), jejunum (mean, 2.3 mm; 95% CI, 2.1 to 2.5 mm), ileum (mean, 2.8 mm; 95% CI, 2.5 to 3.2 mm), and colon (mean, 1.5 mm; 95% CI, 1.4 to 1.7 mm). PMID- 10825094 TI - Evaluation of castor bean toxicosis in dogs: 98 cases. AB - Castor beans (Ricinus communis) contain ricin. Ricin is a glycoprotein reported to cause hypotension, gastroenteritis, depression, and death. However, few deaths are reported following castor bean ingestion in animals. From January 1987 to December 1998, the American Society for the Prevention of Cruelty to Animals National Animal Poison Control Center received 98 incidents of castor bean ingestion in dogs. The most commonly reported clinical signs were vomiting, depression, and diarrhea. Death or euthanasia occurred in 9% of the cases. The severity of clinical signs following castor bean ingestion may depend on whether the beans were chewed or swallowed whole. PMID- 10825095 TI - Intraspinal synovial cyst in a dog. AB - An eight-year-old, male Siberian husky cross was referred with a history of an acute onset of pelvic-limb ataxia and paraparesis. Radiography and subsequent myelography of the spine revealed an extradural compression of the spinal cord at the level of the 13th thoracic (T13) to first lumbar (L1) vertebrae. Hemilaminectomy resulted in the successful removal of an extradural cystic lesion. The morphological diagnosis based on histopathology was a synovial cyst with chondromatosis. There were no postoperative complications, and the dog's condition improved markedly. At two years postoperatively, the animal remains normal on both physical and neurological examination. To the authors' knowledge, this article is the first report of an intraspinal synovial cyst in a dog. PMID- 10825096 TI - Meningoencephalitis secondary to bacterial otitis media/interna in a dog. AB - Central nervous system (CNS) complications of bacterial otitis media/interna are an infrequent occurrence in human patients and have rarely been reported in the veterinary literature. Early recognition of CNS involvement and the use of appropriate diagnostic tests to characterize the nature of the lesion(s) are crucial in determining the best course of treatment. In this paper, the authors describe a dog with bacterial meningoencephalitis secondary to otitis media/interna. PMID- 10825097 TI - Outcome following treatment of vertebral tumors in 20 dogs (1986-1995). AB - Twenty dogs with histopathologically confirmed primary (n=15) or metastatic (n=5) osteosarcoma (n=14) or fibrosarcoma (n=6) of the vertebral column were treated with surgery (n=4), radiation therapy and chemotherapy (n=6), surgery and chemotherapy (n=2), or surgery, radiation, and chemotherapy (n=8). All dogs died due to their disease; 15 died due to local failure, and five died due to nonvertebral metastasis. Overall median survival time was 135 days, with a range of 15 to 600 days. Of the factors evaluated, only postoperative neurological status had a significant influence on outcome by multivariate analysis. This study supports the overall guarded prognosis for dogs with vertebral neoplasia. Better combinations of surgery, chemotherapy, and radiation therapy remain to be defined for this difficult subset of animal cancer. PMID- 10825098 TI - Seminoma with cutaneous metastases in a dog. AB - A 10-year-old Great Pyrenees was presented for anorexia and weight loss. On physical examination, the dog was emaciated and showed a large ulcerated lesion on the right lower lip in addition to an enlarged right testicle. Fine-needle aspiration biopsy of the testicle and surgical biopsy of the lip lesion were performed; the histopathological report was consistent with metastatic seminoma. The diagnostic and therapeutic approach in this unusual metastatic seminoma is presented and compared to the previous literature. A multimodality therapy consisting of surgery and chemotherapy is proposed for the clinical management of metastatic seminoma in dogs. PMID- 10825099 TI - Photodynamic therapy as a treatment for esophageal squamous cell carcinoma in a dog. AB - Intrathoracic esophageal squamous cell carcinoma was diagnosed by endoscopy in an 11-year-old, castrated male Labrador retriever with signs of regurgitation and weight loss. Photodynamic therapy with photofrin was administered three times under endoscopic guidance over a two-month period. A partial response to photodynamic therapy was supported by a reduction in tumor size (noted on serial endoscopic examinations) and by a return to oral alimentation. The dog was euthanized due to recurrent regurgitation and aspiration pneumonia nine months after the onset of therapy. Necropsy revealed marked local invasiveness and regional lymph node metastasis of the esophageal squamous cell carcinoma in addition to pneumonia. The application of photodynamic therapy in the treatment of canine esophageal squamous cell carcinoma is discussed and compared with the human literature. PMID- 10825100 TI - Mineralization of the supraspinatus tendon in dogs: a long-term follow-up. AB - Mineralization of the supraspinatus tendon was diagnosed in 24 large-breed dogs as a probable cause for a chronic unilateral forelimb lameness. Owners of 12 dogs responded to a questionnaire survey evaluating the treatment that their dog had received which consisted of either surgical removal of the mineralization after failure of conservative treatment (operated group; n=9) or rest and nonsteroidal anti-inflammatory drugs (NSAIDs) (nonoperated group; n=3). In eight out of the 12 dogs, the mineralization was also present in the asymptomatic forelimb. Based on owner evaluation, the degree of lameness had decreased distinctly in both groups. Six dogs (four operated and two nonoperated) were reevaluated at Michigan State University Veterinary Teaching Hospital (MSU-VTH) and were without lameness except for one dog in the operated group. The mineralizations had reformed in all dogs in the operated group after a mean follow-up time of 5.1 years. PMID- 10825101 TI - Spontaneous bilateral coxofemoral luxation in four dogs. AB - Spontaneous bilateral coxofemoral luxations and their clinical managements are described in four dogs. Luxations in each case occurred while the patient was in a controlled environment with no possibility of trauma. While all cases had bilateral luxations, the luxations occurred in a staged fashion in two of the four cases. Treatment varied with each case, but included closed reduction, triple pelvic osteotomy, prosthetic capsulorrhaphy, toggle pin, ischioilial pinning, total hip replacement, and femoral head and neck ostectomy. Other than femoral head and neck ostectomy, results were uniformly poor, with reluxation being the most common complication. Evidence of pre-existing degenerative changes was minimal, particularly considering the ages of the patients (range, three to 12 yrs). Histopathological evaluation of joint capsules, round ligaments, and femoral heads are warranted in future cases. PMID- 10825102 TI - Radiologic case study. Synovial osteochondromatosis. PMID- 10825103 TI - Hip instability in the newborn. PMID- 10825104 TI - Neurologic injury from screw displacement. PMID- 10825105 TI - Quality care and cost in Y2K: beating the odds? PMID- 10825106 TI - Surgical approach in THA: the direct lateral approach is more practical and appealing. PMID- 10825107 TI - Is the posterior approach to THA a choice for all seasons? PMID- 10825108 TI - Single-incision technique for internal fixation of distal tibia and fibula fractures. AB - Open reduction and internal fixation of distal tibia and fibula fractures generally involves two separate incisions: an anteromedial incision to approach the tibia and a lateral incision to approach the fibula. Exposing the distal tibia from the medial side is associated with the risk of wound dehiscence, infection, and discomfort to the patient since the hardware is directly beneath the skin. By using a single incision from the anterolateral side, the fibular fracture can be fixed and the lateral aspect of the distal tibia can be safely approached for internal fixation, thus eliminating the need for two separate incisions. PMID- 10825109 TI - Accurate limb-length equalization during total hip arthroplasty. AB - A method of equalization of limb lengths during total hip arthroplasty (THA) was developed that uses the concept of precise reproduction of the position of the femur in space (abduction/adduction) by use of a carpenter's level. Precise reproduction of the femoral position allows accurate measurement of the distance between the pelvis and femur before hip dislocation and after trial component placement allowing accurate measurement of the change in the distance between the femur and pelvis. Accordingly, limb length can be maintained or adjusted to match the contralateral side with increased accuracy. A prospective study was performed with 117 consecutive patients undergoing THA to assess the accuracy of an intraoperative limb-length measuring device. Patients were sequentially randomized into two groups. Group A patients underwent THA without the use of the measuring device, and group B underwent THA using the device. Radiographic assessments of limb lengths were measured using the method of Williamson and Reckling. The hips in group B had a statistically significant decrease in limb length inequality after THA compared with group A (P<.01). Average postoperative limb-length inequality was 8.8 mm and 3.4 mm for groups A and B, respectively. Three (5%) of 58 group B patients and 18 (31%) of 59 group A patients had a radiographic postoperative limb-length inequality >12 mm (P<.01). Eighty-four of group B patients had limb lengths within 6 mm of the contralateral side compared with 30% of group A patients. Twenty-four percent of group A patients and 7% of group B patients had a symptomatic limb-length inequality that required a heel lift (P<.01). PMID- 10825110 TI - Prosthesis cost containment in total joint replacement: a physician-driven free market approach. AB - As an alternative to a limited vendor/volume discount approach, our hospital employed a physician-driven free market strategy aimed at reducing joint implant costs. Surgeons were provided with vendor pricing and peer profile comparisons of implant cost data and asked to select implants providing the best value based on patient need. Vendors were challenged to reduce prices where appropriate. Total savings based on the 1995-1997 volume-adjusted cost difference were $1,059,159, a 17.5% decrease. These results demonstrate the possibility of reducing joint implant costs using a strategy that does not limit vendors or cap prices. PMID- 10825111 TI - Clinical and pathologic characteristics of lumbar disk herniation in the elderly. AB - The clinical and pathologic characteristics of lumbar disk herniation in 23 elderly patients (15 men and 8 women) who required surgical treatment were investigated. Average age at surgery was 67.3 years, and average follow-up was 23 months. Preoperatively, the predominant symptom was severe unilateral leg pain, and 13 patients were nonambulatory because of leg pain. Operative treatment included wide laminectomy in 8, hemilaminectomy in 3, fenestration in 10, and osteoplastic laminectomy in 2 patients. Four (17%) patients had disk herniation at the L2-L3 or L3-L4 level. Sequestered herniation with or without migration was observed in 15 (65%) patients. Of 14 sequestered disk fragments examined histologically, 8 (57%) specimens contained cartilaginous or bony end plate with anulus fibrosus or nucleus pulposus. Postoperatively, results were rated as excellent in 11 patients and good in 12; no patient was rated as fair or poor. Severe leg pain affecting activities of daily life is a predominant symptom of disk herniation in the elderly. For patients in whom conservative treatment has failed, operative treatment should be considered. In lumbar disk herniation in the elderly, the incidence of cephalad and lateral herniation is higher than in younger patients. In addition, sequestered and migrated herniations including end plate are frequent. PMID- 10825112 TI - Surgical treatment of comminuted intra-articular fractures of the distal humerus with double tension band osteosynthesis. AB - Intra-articular fractures of the distal humerus are usually treated with cancellous screws for the condylar metaphysis itself and two Y-shaped plates for fixation of the metaphysis to the shaft. Twenty-four patients with comminuted intra-articular fractures (class C on AO/ASIF classification) were treated with crossed K-wires for the condylar metaphysis and double tension band osteosynthesis for fixation of the metaphysis block to the shaft. Excellent or good results were achieved in 83% of patients. This technique is easier to perform and more cost effective than most other fixation methods. Additionally, it offers good stability and allows for earlier functioning of the elbow. Complication rates are comparable to other methods of internal fixation. PMID- 10825113 TI - A biomechanical evaluation of internal fixation for ankle arthrodesis comparing two methods of joint surface preparation. AB - This study compared the biomechanics of two methods of joint preparation for ankle fusion with internal fixation. In the first method, the articular cartilage of the ankle joint was removed leaving the joint contour intact. In the second method, parallel flat saw cuts were used to prepare the joint surface for fusion. The specimens were loaded submaximally in a cyclic repetitive fashion before they were loaded to failure. Increasing joint motion occurred as the number of cycles increased. The difference between the two specimen types was not statistically significant during the repetitive tests or when loaded to failure. These results demonstrate both techniques of joint preparation studied are useful and biomechanically sound. PMID- 10825114 TI - Early results of isoelastic hemiarthroplasty in chronic shoulder arthritis. AB - Fourteen isoelastic shoulder hemiarthroplasties were studied in patients with shoulder pain due to chronic rheumatoid arthritis and osteoarthritis unresponsive to conservative means. The average follow-up was 27 months (range: 12-46 months) and the average patient age was 66.4 years (range: 54-79 years). A Constant shoulder score was used to assess results. Pain relief was obtained in 85.7% of patients. Function scores averaged 8 preoperatively and 13 postoperatively. Subjectively, 12 patients improved. Average gain was 33 degrees of abduction, 35 degrees of flexion, and 21 degrees of external rotation. Internal rotation improved by two levels on the spine. Total shoulder scores improved from an average of 42 points preoperatively to 78 points postoperatively. There was 1 complication of immediate postoperative dislocation. Early results with isoelastic (polyacetyl resin) shoulder hemiarthroplasty are encouraging. The prosthesis is inexpensive and easy to use, requires minimal bone resection, involves cementless fixation, and has established results in tumor surgery. Its long-term value for chronic arthritis will be reported as part of an ongoing prospective study. PMID- 10825115 TI - Transfusion in unilateral total knee arthroplasty. AB - Blood transfusion requirements were analyzed in 102 total knee arthroplasties in 92 patients. All components were cemented, and no postoperative drainage was used. Six patients received transfusions; two of these patients received transfusions for symptoms and four received transfusions prophylactically. These findings do not support the routine practice of preoperative autologous blood donation. PMID- 10825116 TI - An investigation into the role of oxygen free radical scavengers in preventing polymethylmethacrylate-induced necrosis in an osteoblast cell culture. AB - This study examined the effect of polymethylmethacrylate (PMMA) on osteocytic necrosis and the role of free radical scavengers in minimizing this damage. Bovine osteoblast cells with a characteristic phenotype were seeded at a density of 4x10(4) cells/cm2 and cultured in a DMEM supplemented with 10% fetal calf serum. A transwell insert with 2 cc of PMMA was suspended above the culture, and a time log response curve was established following elusion of free radicals around the osteoblast media. Chemiluminescence was used to determine quantitative free radical release. Using a Student's two-tailed t test there was a significant difference in the amount of hydroxyl radical released at 1-6 hours compared with controls (P=.028). Using histologic markers, there was a significant correlation between the use of PMMA and osteoblast cell necrosis. Transwell plates were coated with varying concentrations of mannitol, a known hydroxyl radical scavenger. A log dose response curve was established. There was a clear statistical association between a 10% mannitol solution and a reduction in the free radical release from PMMA (P=.03). Similarly, using Trypan blue histologic staining, there was a significant reduction in PMMA-induced cell necrosis when 10% mannitol was used as a scavenger (P=.01). A Rockwell superficial hardness test was used to determine whether mannitol had any effect on the surface hardness of the polymer. No statistical difference could be found between those treated with mannitol and controls at a depth of up to 1 mm. These results demonstrate hydroxyl radical is released from the polymerization reaction of PMMA. These radicals cause cell death in an osteoblast culture medium. This has been addressed using a 10% mannitol solution, which reduced cell necrosis. PMID- 10825117 TI - Femoral head and neck dissociation after a total hip arthroplasty with a constrained acetabular liner. PMID- 10825118 TI - Enchondroma versus chondrosarcoma of the phalanx. PMID- 10825119 TI - The Mennen plate in complex humeral fractures. PMID- 10825120 TI - Giant juxtacortical chondrosarcoma of the humerus. PMID- 10825121 TI - Ultra high molecular weight polyethylene wear in total hip arthroplasty. PMID- 10825122 TI - Immunoprevention of cancer: is the time ripe? AB - Immunotherapy applied to patients with established tumors rarely leads to an objective response, whereas patients apparently free from disease after conventional treatment and at risk of recurrence are beginning to receive vaccination. New classes of patients or not-yet patients are those with a high genetic or environmental risk of developing cancer. They may draw benefit from a "soft" treatment such as vaccination. This overview discusses the prospects of immune stimulation as a means of cancer prevention by inducing various forms of nonspecific or even specific immunity. Attainment of this goal provides the rationale and motivation for embarking on such a new and potentially rewarding enterprise. PMID- 10825123 TI - Effects of the multidrug transporter P-glycoprotein on cellular responses to ionizing radiation. AB - Ionizing radiation induces apoptosis, mitotic catastrophe, and senescence-like terminal proliferation arrest in tumor cells. We investigated the effect of the MDR1 P-glycoprotein (Pgp), recently shown to inhibit caspase-mediated apoptosis, on cellular responses to radiation. Pgp strongly inhibited radiation-induced apoptosis in a HeLa-derived cell line with inducible MDR1 expression and in NIH 3T3 cells transduced with a MDR1-expressing retroviral vector. The inhibition of apoptosis by Pgp was associated, however, with increases in radiation-induced mitotic catastrophe and senescence and produced only a marginal change in the survival of irradiated cells. Pgp had no effect on radiation responses in apoptosis-resistant HT1080 cells. These results indicate that Pgp inhibits radiation-induced apoptosis, but this effect of Pgp provides no substantial increase in radiation resistance of the tested cell lines because apoptosis resistant cells die from mitotic catastrophe or undergo senescence-like terminal proliferation arrest. PMID- 10825124 TI - Influence of nitric oxide synthase II gene disruption on tumor growth and metastasis. AB - The relationship between nitric oxide (NO) synthase II (NOS II) expression and the metastatic ability of tumor cells is inconclusive. We determined the role of host NOS II expression in the growth and metastasis of the B16-BL6 murine melanoma and M5076 murine ovarian sarcoma cell lines. The cells were either s.c. or i.v. injected into syngeneic wild-type (NOS H+/+) and NOS II-null (NOS H-/-) C57BL/6 mice. Both cell lines produced slightly larger s.c. tumors in NOS H-/- mice than in NOS II+/+ mice. However, B16- BL6 cells produced more and larger experimental lung metastases in NOS II+/+ mice than in NOS II-/- mice, whereas M5076 cells produced fewer and smaller experimental lung metastases in NOS II+/+ mice than in NOS II-/- mice. After activation with IFN-gamma and lipopolysaccharide, macrophages isolated from NOS II+/+ C57BL/6 mice produced NO dependent cytotoxicity in sarcoma cells, whereas macrophages from NOS II-/- C57BL/6 mice did not. In contrast, activated macrophages produced little to no NO mediated cytotoxicity in melanoma cells. Immunostaining analyses indicated that NOS II expression was apparent in the metastases growing in NOS H+/+ mice and correlated with increased cell proliferation in B16-BL6 lung metastases but with decreased cell proliferation in M5076 liver metastases. Our data suggest that disruption of host NOS II expression enhanced the growth and metastasis of NO sensitive tumor cells but suppressed the metastasis of NO-resistant tumor cells, proposing that host-derived NO may differentially modulate tumor progression. PMID- 10825125 TI - Effects of Thomsen-Friedenreich antigen-specific peptide P-30 on beta-galactoside mediated homotypic aggregation and adhesion to the endothelium of MDA-MB-435 human breast carcinoma cells. AB - Both the ability of malignant cells to form multicellular aggregates via homotypic or heterotypic aggregation and their adhesion to the endothelium are important if not critical during early stages of cancer metastasis. The tumor associated carbohydrate Thomsen-Friedenreich antigen (T antigen) and beta galactoside binding lectins (galectins) have been implicated in tumor cell adhesion and tissue invasion. In this study, we demonstrate the involvement of T antigen in both homotypic aggregation of MDA-MB-435 human breast carcinoma cells and their adhesion to the endothelium. The T antigen-specific peptide P-30 (HGRFILPWWYAFSPS) selected from a bacteriophage display library was able to inhibit spontaneous homotypic aggregation of MDA-MB-435 cells up to 74% in a dose dependent manner. Because T antigen has beta-galactose as a terminal sugar, the expression profile of beta-galactoside-binding lectins (galectins) in MDA-MB-435 cells was studied. Our data indicated the abundant expression of [35S]methionine/cysteine-labeled galectin-1 and galectin-3 in this cell line, which suggested possible interactions between galectins and T antigen. As revealed by laser confocal microscopy, both galectin-1 and galectin-3 also participate in the adhesion of the MDA-MB-435 cells to the endothelium. We observed the clustering of galectin-3 on endothelial cells at the sites of the contact with tumor cells, consistent with its possible interaction with T antigen on cancer cells The galectin-1 signal, however, strongly accumulated at the sites of cell-cell contacts predominantly on tumor cells. The T antigen-specific P-30 significantly (50%) inhibited this adhesion, which indicated that T antigen participates in the adhesion of MDA-MB-435 breast cancer cells to the endothelium. The ability of synthetic P-30 to inhibit both the spontaneous homotypic aggregation of MDA-MB-435 cells and their adhesion to the endothelium (>70 and 50%, respectively) suggests its potential functional significance for antiadhesive therapy of cancer metastasis. PMID- 10825126 TI - Breast cancer resistance protein is localized at the plasma membrane in mitoxantrone- and topotecan-resistant cell lines. AB - Tumor cells may display a multidrug resistant phenotype by overexpression of ATP binding cassette transporters such as multidrug resistance (MDRI) P-glycoprotein, multidrug resistance protein 1 (MRP1), and breast cancer resistance protein (BCRP). The presence of BCRP has thus far been reported solely using mRNA data. In this study, we describe a BCRP-specific monoclonal antibody, BXP-34, obtained from mice, immunized with mitoxantrone-resistant, BCRP mRNA-positive MCF-7 MR human breast cancer cells. BCRP was detected in BCRP-transfected cells and in several mitoxantrone- and topotecan-selected tumor cell sublines. Pronounced staining of the cell membranes showed that the transporter is mainly present at the plasma membrane. In a panel of human tumors, including primary tumors as well as drug-treated breast cancer and acute myeloid leukemia samples, BCRP was low or undetectable. Extended studies will be required to analyze the possible contribution of BCRP to clinical multidrug resistance. PMID- 10825127 TI - Estradiol induces functional inactivation of p53 by intracellular redistribution. AB - Estrogen treatment of MCF-7 cells grown in serum-free medium induced a modification of the intracellular distribution of p53 protein. Western blot analysis and immunofluorescence staining showed that p53 was localized in the nucleus of untreated cell and that after 48 h of hormone treatment, it was mostly localized in the cytoplasm. This effect was blocked by the antiestrogen ICI182,780. Intracellular redistribution of p53 was correlated to a reduced expression of the WAF1/CIP1 gene product and to the presence of degradation fragments of p53 in the cytosol. Estradiol treatment prevented the growth inhibition induced by oligonucleotide transfection, simulating DNA damage. This observation indicated that the wild-type p53 gene product present in the MCF-7 cell could be inactivated by estradiol through nuclear exclusion to permit the cyclin-dependent phosphorylation events leading to the G1-S transition. In addition, the estradiol-induced inactivation of p53 could be involved in the tumorigenesis of estrogen-dependent neoplasm. PMID- 10825128 TI - Monoallelic amplification of estrogen receptor-alpha expression in breast cancer. AB - Gene amplification and loss of heterozygosity are alterations to chromosomal structure whereby tumor cells alter patterns of gene expression. We have identified a novel mechanism of gene regulation in which cancer cells predominantly express one of the two alleles of a gene. Estrogen receptor (ER) alpha is overexpressed in hormone-responsive breast cancer compared with normal breast epithelial cells. Using a polymorphism of codon 10, we examined allele specific expression of the four different ER promoters in MCF-7 breast cancer cells and primary tumors. Monoallelic amplification of expression (MAX) for all four ER promoters was identified, resulting in an allelic preference of > 100 fold. MAX was the result of an amplification of allele copy number and a preference to transcribe the amplified allele. The effect of MAX was most significant for the promoters clustered near the 1' exon, whereas the expression from the distant H promoter mirrored template copy number. MAX of the ER gene was not found to occur in normal endometrial or breast tissue. As a novel mechanism in cancer genetics, MAX can result in functional homozygosity at a gene locus. PMID- 10825129 TI - A novel transmembrane serine protease (TMPRSS3) overexpressed in pancreatic cancer. AB - We report the characterization of a novel serine protease of the chymotrypsin family, recently isolated by cDNA-representational difference analysis, as a gene overexpressed in pancreatic cancer. The 2.3-kb mRNA of the gene, named TMPRSS3, is strongly expressed in a subset of pancreatic cancer and various other cancer tissues, and its expression correlates with the metastatic potential of the clonal SUIT-2 pancreatic cancer cell lines. The deduced polypeptide sequence consists of 437 amino acids and exhibits all of the structural features characteristic of serine proteases with trypsin-like activity. TMPRSS3 is membrane bound with a NH2-terminal signal-anchor sequence and a glycosylated extracellular region containing the serine protease domain. Thus, TMPRSS3 is a novel membrane-bound serine protease overexpressed in cancer, which may be of importance for processes involved in metastasis formation and tumor invasion. PMID- 10825130 TI - Excision of tamoxifen-DNA adducts by the human nucleotide excision repair system. AB - The antiestrogen tamoxifen is used in the treatment of breast cancer and has recently been recommended as a chemopreventive drug for women at high risk for breast cancer. However, women treated with the drug have an increased incidence of endometrial cancer. It has been suggested that this endometrial cancer might result from mutagenic DNA adducts, which are formed by electrophilic tamoxifen species generated by metabolic activation of the drug. Because the frequency of damage-induced mutations is strongly dependent on the repairability of the lesion, we investigated the repair of the major tamoxifen-DNA adducts by the human nucleotide excision repair system. Using the reconstituted human excision repair system and synthetic DNA substrates, we found that the four types of tamoxifen-DNA adducts detected in the endometrium were repaired with moderate to poor efficiency by nucleotide excision repair. It is concluded that individual variations in repair capacity may play a role in the development of tamoxifen induced endometrial cancer. PMID- 10825131 TI - Radiation and the Apo2L/TRAIL apoptotic pathway preferentially inhibit the colonization of premalignant human breast cells overexpressing cyclin D1. AB - The role of cyclin D1 overexpression in human breast premalignancy was investigated using immortal, nontumorigenic MCF-10A cells. Previous work documented that cyclin D1 overexpression promoted in vitro anchorage-independent colonization. We now report that the colonization of MCF-10A cyclin D1 transfectants was preferentially inhibited by gamma-radiation and specific classes of apoptosis inducers [Apo-2 ligand (Apo-2L), but not tumor necrosis factor alpha]. Antibody inhibition studies and semiquantitative PCR indicated that radiation inhibition of colonization was partially mediated via the Apo2L/TRAIL pathway. The apoptotic removal of cyclin D1-overexpressing, colonization-competent premalignant breast cells by Apo2L/TRAIL or other biologicals may represent a novel approach to the prevention of breast cancer. PMID- 10825132 TI - p16INK4A mediates cyclin dependent kinase 4 and 6 inhibition in senescent prostatic epithelial cells. AB - The senescence checkpoint constrains the proliferative potential of normal cells in culture to a finite number of cell doublings. In this study, we investigated the mechanism of cyclin dependent kinase (cdk) inhibition in senescent human prostatic epithelial cells (HPECs). Progression of HPECs from early passage to senescence was accompanied by a gradual loss of cells in S phase and an accumulation of cells containing 2N DNA. Furthermore, G1-S phase-associated kinase activities progressively diminished with increasing cell passage. In senescent HPECs, cdk4 and cyclin E1- and A-associated kinases were catalytically inactive. In contrast to observations in senescent fibroblasts, levels of the kinase inhibitor protein (KIP) inhibitor p21CIP1 diminished over the proliferative life span of HPECs. p27KIP1 levels fell as cells approached senescence, and the association of both p21CIP1 and p27KIP1 with cdk4/6 complexes was decreased. However, the level of cyclin E1-associated KIP molecules was unaltered as cells progressed into senescence. Progression to senescence was accompanied by a progressive increase in both the level of p16(INK4A) and in its association with cdk4 and cdk6. As HPECs approached senescence, cdk4- and cdk6 bound p16(INK4A) showed a shift to a slower mobility due to a change in its phosphorylation profile. As p16(INK4A) increased in cdk4 and cdk6 complexes, there was a loss of cyclin D1 binding. The altered phosphorylation of p16(INK4A) in senescent prostatic epithelial cells may facilitate its association with cdk4 and cdk6 and play a role in the inactivation of these kinases. PMID- 10825133 TI - High and low fluences of alpha-particles induce a G1 checkpoint in human diploid fibroblasts. AB - The effects of exposure to high and very low fluence alpha-particles on the G1 checkpoint were investigated in human diploid fibroblasts irradiated and released from density-inhibited confluent cultures by the use of the cumulative labeling index method. Transient and permanent arrests in G1 occurred in fibroblast populations exposed to mean doses as low as 1 cGy, suggesting that nontraversed bystander cells may contribute to the low dose response. In cells exposed to high fluences, the G1 checkpoint is at least as extensive as in gamma-irradiated cells. In contrast to gamma-irradiated cells, neither repair of potentially lethal damage nor a reduction in the fraction of cells transiently or permanently arrested in G1 were observed in cells held in confluence for 6 h after alpha particle irradiation. Studies with isogenic wild-type, p53-/-, and p21Waf1-/- mouse embryo fibroblasts exposed to either gamma or alpha-particle radiation revealed a total lack of G1 arrest in either p53-/- or p21waf1-/- cells, indicating that the G1 checkpoint in wild-type cells is p53-dependent and that p21Wf1 fully mediates the role of p53 in its induction. In contrast to human cells, mouse embryo fibroblasts do not undergo a permanent G1 arrest. Except under conditions favoring potentially lethal damage repair, a comparable expression pattern of p53, p21Waf1, and other cell cycle-regulated proteins (pRb, p34cdc2, and cyclin B1) was observed in alpha-particle or gamma-irradiated human fibroblasts. PMID- 10825134 TI - Clinical implications of expression of interleukin 8 related to angiogenesis in uterine cervical cancers. AB - There was a significant correlation between microvessel counts and interleukin (IL)-8 levels and between infiltrated macrophage counts and IL-8 levels in uterine cervical cancers. Immunohistochemical staining revealed that the localization of IL-8 was similar to that of CD68 for macrophages. The prognosis of the 20 patients with high IL-8 (>1000 pg/mg protein) in uterine cervical cancers was extremely poor, whereas the 24-month survival rate of the other 60 patients with low IL-8 (<1000 pg/mg protein) was 67%. Therefore, this indicates that IL-8 might be a prognostic indicator as an angiogenic factor supplied from macrophages within and around the tumor. PMID- 10825135 TI - Depletion of tumor oxygenation during photodynamic therapy: detection by the hypoxia marker EF3 [2-(2-nitroimidazol-1[H]-yl)-N-(3,3,3 trifluoropropyl)acetamide ]. AB - Photodynamic therapy (PDT) of tumors can create hypoxia when oxygen is depleted by photochemical consumption or the oxygen supply is compromised by microvascular damage. However, oxygen is a requirement for PDT, and hypoxia during illumination can lead to poorer tumor response. As such, sensitive methods of quantifying tumor oxygen and evaluating its distribution may help in the development and optimization of treatment protocols. In this study, the hypoxia marker EF3 [2-(2 nitroimidazol-1[H]-yl)-N-(3,3,3-trifluoropropyl)acetam ide] was used to evaluate the oxygenation of PDT-treated radiation-induced fibrosarcoma tumors. Tumor bearing mice were administered Photofrin (5 mg/kg) 24 h before PDT illumination at 75 mW/cm2, 135 J/cm2 (30 min). EF3 (52 mg/kg) was injected either within 3 min before PDT illumination, with tumor excision at the conclusion of illumination, or within 3 min after illumination, with tumor excision 30 min later. Control animals received EF3 alone, EF3 plus Photofrin, or EF3 plus illumination. After tumor disaggregation, staining with a fluorochrome-conjugated monoclonal antibody, and flow cytometric analysis, control tumors demonstrated an averaged median fluorescence intensity (+/- SE) of 17.1 +/- 2.8. EF3 binding significantly (P = 0.007) increased during PDT to a median fluorescence intensity of 48.9 +/- 8.3. In the 30 min after PDT, EF3 binding returned to control levels (median, 18.3 +/- 3.3). To evaluate the oxygen concentrations corresponding to these fluorescence intensities, an in vitro standard curve was created based on the in vivo exposure conditions. From this curve, the oxygen tensions of tumors exposed to EF3 under control conditions, during PDT, or after PDT were calculated to be 3.1-5.3, 1.2-2.4, and 3.0-5.2 mm Hg, respectively. Detection of EF3 binding using a monoclonal antibody correlated well with direct detection of binding using a radioactive assay. EF3 binding was linear with drug incubation for times from 1.5 to 60 min. Overall, this work demonstrates that hypoxia during PDT illumination of radiation-induced fibrosarcoma tumors can be detected by the hypoxia marker EF3. Hypoxia during illumination can be labeled separately from that found before or after PDT. Tissue oxygen tensions corresponding to EF3 binding levels can be calculated. PMID- 10825136 TI - Fas-dependent and -independent mechanisms of cell death following DNA damage in human colon carcinoma cells. AB - In thymidylate synthase-deficient (TS-) colon carcinoma cells, thymineless death is mediated via Fas/Fas ligand (FasL) interactions after thymidine deprivation and inhibited by the Fas-inhibitory monoclonal antibody NOK-1. The objective of the study was to elucidate whether other modes of DNA damage induced by doxorubicin, topotecan, and etoposide (VP-16) could elicit a similar cytotoxic response in TS- cells by signaling via the Fas death receptor. After a 72-h drug exposure, a loss in clonogenic survival that was not prevented by NOK-1 was induced by each agent in the absence of acute apoptosis, yielding IC50 values of 5 (doxorubicin), 10 (topotecan), and 150 nM (VP-16). Furthermore, TS- cell clones selected for resistance to Fas-mediated apoptosis (CH-11) were cross-resistant to the induction of thymineless death after thymidine deprivation but were not cross resistant to doxorubicin, topotecan, or VP-16. A close correlation was found between acute induction of apoptosis (24 h) and up-regulated expression of FasL at high concentrations of each of the three agents (0.3-3 microM doxorubicin, 0.3 3 microM topotecan, and 10-90 microM VP-16), which was caspase dependent but Fas independent. At all drug concentrations, cell cycle distribution analyses demonstrated marked accumulation of cells in the G2-M phase. At nanomolar drug concentrations, prolonged arrest of TS- cells in G2-M phase resulted in the up regulation of FasL expression and the delayed appearance of apoptotic cells (6 days), which could also be inhibited by the general caspase inhibitor Z-VAD-FMK, but not by NOK-1 or Fas-Fc. In clonogenic assays, Z-VAD-FMK did not rescue cells treated with VP-16 in contrast to treatment with CH-11 or thymineless stress, suggesting an irreversible commitment to cell death in G2-M phase. Expression of FasL at all drug concentrations appeared to be unrelated to the mechanism of drug induced apoptosis. This was in contrast to the Fas-dependent regulation of thymineless death, which could be inhibited by blocking Fas/FasL interactions. PMID- 10825137 TI - Induction of apoptosis in leukemic cells by the reversible microtubule-disrupting agent 2-methoxy-5-(2',3',4'-trimethoxyphenyl)-2,4,6-cycloheptatrien-1 -one: protection by Bcl-2 and Bcl-X(L) and cell cycle arrest. AB - We have found that the bicyclic colchicine analogue 2-methoxy-5-(2',3',4' trimethoxyphenyl)-2,4,6-cycloheptatrien-1-on e (MTC) induced a dose- and time dependent apoptotic response in human leukemic cells. MTC and colchicine rapidly disrupted the microtubule integrity and arrested cells at the G2-M phase before the onset of apoptosis. These responses were mediated by microtubule inhibition because 2-methoxy-5-[[3-(3,4,5-trimethoxyphenyl)propionyl]amino]-2,4,6-cycloh eptatrien-1-one and lumicolchicine, inactive analogues of MTC and colchicine, respectively, were unable to promote microtubule disassembly, cell cycle arrest, and apoptosis. Although 1 microM MTC induced a complete microtubule disruption after 1 h of incubation in human leukemic HL-60 cells that led to an accumulation of cells at the G2-M phase, MTC-induced apoptosis occurred after 9 h of treatment. This indicates the existence of a rather long lag between microtubule disruption and the onset of apoptosis. Unlike colchicine, the removal of MTC during this lag resulted in rapid microtubule repolymerization, followed by restoration of normal cell cycle and cell growth. MTC, but not 2-methoxy-5-[[3 (3,4,5-trimethoxyphenyl)-propionyl]amino]-2,4,6-cyclo heptatrien-1-one, induced c jun expression as well as c-Jun NH2-terminal kinase and caspase activation, indicating that these signaling pathways are triggered by the specific action of MTC on microtubules. Caspase inhibition prevented MTC-induced apoptosis. Overexpression of bcl-2 or bcl-xL by gene transfer in human erythroleukemic HEL cells abrogated MTC-induced apoptosis, but cells remained arrested in G2-M, suggesting that bcl-2 and bcl-xL block the signaling pathway between G2-M arrest and triggering of apoptosis. MTC-treated bcl-2 and bcl-xL-transfected HEL cells recovered their capacity to proliferate after MTC removal. These results indicate that microtubule inhibition induces G2-M arrest and apoptosis in leukemic cells, showing a lag phase between G2-M arrest and the onset of apoptosis, regulated by bcl-2 and bcl-xL, during which MTC displays a reversible action on microtubule depolymerization and G2-M cell cycle arrest. Thus, MTC is a potent apoptotic inducer on human leukemic cells and shows a remarkable reversible action on microtubule network and cell cycle before commitment for apoptosis is reached. PMID- 10825138 TI - Liposome-delivered angiostatin strongly inhibits tumor growth and metastatization in a transgenic model of spontaneous breast cancer. AB - The possibility to inhibit tumor growth by interfering with the formation of new vessels, which most neoplasias depend on, has recently raised considerable interest. An angiogenic switch, in which proliferating cells acquire the ability to direct new vessel formation, is thought to be an early step in the natural history of solid tumors. Using a transgenic model of breast cancer, which shows many similarities to its human counterpart, including ability to metastasize, we targeted angiostatin production to an early stage of tumor formation. Liposome delivered angiostatin considerably delayed primary tumor growth and, more importantly, inhibited the appearance of lung metastases. These findings can be relevant to the design of therapeutic intervention in humans. PMID- 10825139 TI - Efficient induction of cell death by adenoviruses requires binding of E1B55k and p53. AB - The use of an Elb55k-deficient adenovirus, ONYX-015, to selectively target tumor cells containing a mutated p53 gene has produced promising results. However, recent reports have questioned the selectivity of this virus, showing that ONYX 015 can replicate in cells containing a wild-type p53 and that p53 may actually be required for cell death. To address these apparent contradictions in the literature, we infected a number of mutant and wild-type p53-containing cell lines with ONYX-015 and wild-type adenovirus and observed their death profiles up to 10 days postinfection. We demonstrate that two distinct cell death phenotypes exist, one of which is rapid and dependent on the presence of p53 and one of which is p53 independent. Using adenoviruses expressing E1b55k proteins deficient in their ability to bind p53, we show that formation of a complex between p53 and the adenoviral Elb55k protein is necessary for the activation of the rapid cell death pathway. In the absence of p53 or the absence of complex formation between p53 and Elb55k, cell death is delayed considerably. These data suggest three things: that the selectivity of killing appears to be dependent on the presence of the E1b55k/p53 complex; that viruses lacking Elb55k (such as ONYX-015) kill cells in a delayed manner independent of p53; and that binding of E1b55k to p53 does not merely serve to inactivate p53, but rather is required for the induction of rapid cell death. The components of this complex that lead to rapid cell death remain to be determined. PMID- 10825140 TI - Two novel 14-Epi-analogues of 1,25-dihydroxyvitamin D3 inhibit the growth of human breast cancer cells in vitro and in vivo. AB - The biological activity of two novel 14-epi-analogues of 1,25(OH)2D3, 19-nor-14 epi-23-yne-1,25(OH)2D3 (TX 522) and 19-nor-14,20-bisepi-23-yne-1,25(OH)2D3 (TX 527), is described. Both analogues were at least 10 times more potent than 1,25(OH)2D3 in inhibiting in vitro cell proliferation and had much lower in vivo calcemic effects than 1,25(OH)2D3. Treatment with 1,25(OH)2D3, TX 522, or TX 527 in vitro was accompanied by an accumulation of cells in the G1 phase of the cell cycle. Protein levels of cyclin C and cyclin D1 in in vitro cultures of MCF-7 cells were down-regulated to 50 and 30%, respectively, of control levels at 72 and 120 h after stimulation. Protein levels of p21 and p27 at 72 h were significantly enhanced by 1,25(OH)2D3 and TX 522 but surprisingly not by TX 527. The inability of TX 527 to up-regulate p21 seemed to be cell type specific because p21 was induced in other cell types. Diminished phosphorylation of the retinoblastoma protein after treatment with 1,25(OH)2D3, TX 522, or TX 527 may ultimately contribute to the growth inhibition caused by these compounds. According to the data presented, the induction of apoptosis seemed not to be a major mechanism responsible for the growth-inhibitory effect of 1,25(OH)2D3 and analogues. Both 14-epianalogues significantly retarded tumor progression (40% reduced compared with control mice) in an in vivo model of MCF-7 breast cancer cells established in nude mice. In conclusion, these novel analogues have the eligible profile to be tested as therapeutic agents for the treatment of hyperproliferative diseases such as breast cancer. PMID- 10825141 TI - Mouse mammary tumor virus-Ki-rasB transgenic mice develop mammary carcinomas that can be growth-inhibited by a farnesyl:protein transferase inhibitor. AB - For Ras oncoproteins to transform mammalian cells, they must be posttranslationally modified with a farnesyl group in a reaction catalyzed by the enzyme farnesyl:protein transferase (FPTase). Inhibitors of FPTase have therefore been developed as potential anticancer agents. These compounds reverse many of the malignant phenotypes of Ras-transformed cells in culture and inhibit the growth of tumor xenografts in nude mice. Furthermore, the FPTase inhibitor (FTI) L-744,832 causes tumor regression in mouse mammary tumor virus (MMTV)-v-Ha-ras transgenic mice and tumor stasis in MMTV-N-ras mice. Although these data support the further development of FTIs, it should be noted that Ki-ras is the ras gene most frequently mutated in human cancers. Moreover, Ki-RasB binds more tightly to FPTase than either Ha- or N-Ras, and thus higher concentrations of FTIs that are competitive with the protein substrate may be required to inhibit Ki-Ras processing. Given the unique biochemical and biological features of Ki-RasB, it is important to evaluate the efficacy of FTIs or any other modulator of oncogenic Ras function in model systems expressing this Ras oncoprotein. We have developed strains of transgenic mice carrying the human Ki-rasB cDNA with an activating mutation (G12V) under the control of the MMTV enhancer/promoter. The predominant pathological feature that develops in these mice is the stochastic appearance of mammary adenocarcinomas. High levels of the Ki-rasB transgene RNA are detected in these tumors. Treatment of MMTV-Ki-rasB mice with L-744,832 caused inhibition of tumor growth in the absence of systemic toxicity. Although FPTase activity was inhibited in tumors from the treated mice, unprocessed Ki-RasB was not detected. These results demonstrate the utility of the MMTV-Ki-rasB transgenic mice for testing potential anticancer agents. Additionally, the data suggest that although the FTI L-744,832 can inhibit tumor growth in this model, Ki-Ras may not be the sole mediator of the biological effects of the FTI. PMID- 10825142 TI - Recombinant adenovirus vaccine encoding a chimeric T-cell antigen receptor induces protective immunity against a T-cell lymphoma. AB - Vaccination using recombinant tumor-derived T-cell antigen receptor (TCR) protein induces a protective, idiotype-specific immune response against a murine T-cell tumor. However, the technically demanding task of producing patient-specific, recombinant TCR protein restricts the translation of TCR vaccines for clinical use. We report here the development of an effective recombinant TCR adenovirus vaccine. Individual adenoviruses were constructed to encode a chimeric TCR derived from either tumor Valpha or Vbeta regions fused to xenogeneic human constant regions. Coinjection of the chimeric alpha- and the beta-TCR adenoviruses protected mice against tumors. The level of protection was comparable to that achieved by an optimized regimen of recombinant TCR protein vaccines. Tumor immunity induced by TCR adenoviruses required the xenogeneic constant regions and was mediated by CD8+ T cells. Independent vaccines consisting of adenovirus expressing either chimeric alpha- or beta-TCR chain also stimulated a protective immune response. Immunization with TCR adenovirus may offer a new efficacious, protein-free vaccination approach for the treatment of T cell malignancies. PMID- 10825143 TI - Methioninase gene therapy of human cancer cells is synergistic with recombinant methioninase treatment. AB - Results obtained over the past 40 years have demonstrated that tumor cells of all types tested have an elevated growth requirement for methioninase compared with normal cells. Recombinant methioninase (rMETase) cloned from Pseudomonas putida has been found previously to be an effective antitumor agent attributable to deprivation of the extracellular methionine source of the tumor. To degrade intracellular methioninase, we have now developed an adenoviral vector inserted with the P. putida methioninase (MET) gene (rAd-MET). The in vitro efficacy of rAd-MET was tested on the OVCAR-8 human ovarian cancer cell line, the HT1080 human fibrosarcoma cell line, and human normal fibroblasts. rAd-MET transduction of OVACAR-8 and HT1080 resulted in high levels of methioninase expression up to 10% or more of the total protein of the cells, depending on the multiplicity of infection. The IC50 of rAd-MET for OVCAR-8 cells in 96-well plates was approximately 2 x 106 plaque-forming units (pfu)/well. The IC50 of control adenovirus (control-rAd) was 4 x 10(7) pfu/well, 20 times higher than rAd-MET. In the presence of the IC50 of 2 x 10(6) pfu/well of rAd-MET, the addition of 0.025 units/ml of rMETase, which is 25% of the IC50, resulted in a 90% inhibition of tumor cell number. This indicated that rAd-MET enhanced the efficacy of rMETase. In contrast, 2 x 10(6) pfu/well of control-rAd in combination with 0.025 units/ml of rMETase had an efficacy of only 10% inhibition of cell number. The synergistic effect of the combination of rMETase and rAd-MET was quantitated by calculating the combination index (CI). The CIs for all combinations of rAd-MET and rMETase tested on OVCAR-8 were <0.7 with a mean of 0.5, indicating synergy. Similar synergy of rAd-MET and rMETase was seen on HT1080 human fibrosarcoma cells with a mean of 0.74. In contrast, the CIs of all combinations of rMETase and control adenovirus concentrations tested on both cell lines had a mean CI of approximately 1, which indicated that this combination had only an additive effect. The normal fibroblasts, on the other hand, appeared relatively resistant to the MET gene because in the presence of rMETase, 2.5 x 10(7) pfu/well of rAd MET or control rAd had almost an identical effect on cell survival. The selectively strong synergy of rAd-MET and rMETase on cancer cells allows reduced levels of each agent to be used, thus decreasing potential side effects. PMID- 10825144 TI - Endogenous interleukin-18 modulates immune escape of murine melanoma cells by regulating the expression of Fas ligand and reactive oxygen intermediates. AB - It has been known that melanoma cells can suppress the immune system by the Fas ligand. The present study investigated whether interleukin (IL)-18, which can enhance Fas ligand expression, is produced by B16F10 melanoma cells and is involved in immune escape of tumor cells. Immunohistology, reverse transcription PCR, intracellular fluorescence-activated cell-sorting analysis, and immunoblotting demonstrated that melanoma cells express IL-18. C57BL/6 splenocytes cultured with culture supernatants of B16F10 melanoma cells enhanced IFN-gamma production, which was blocked by anti-IL-18 antibody, indicating that IL-18 in the culture supernatants is functional. In addition to IL-18, the IL-18 receptor was also detected in B16F10 melanoma cells, suggesting a role of this cytokine in regulating the functions of B16F10 melanoma cells. The functional effect of IL-18 on B16F10 melanoma cells was shown by reduction of Fas ligand expression in cells treated with anti-IL-18 antibody or transfected with IL-18 antisense cDNA. In addition, the same treatments decreased intracellular reactive oxygen intermediate levels in B16F10 melanoma cells, indicating that IL-18 regulates reactive oxygen intermediate production, which is involved in Fas ligand expression. Furthermore, transfection of IL-18 antisense cDNA into melanoma cells increased the susceptibility of tumor cells to natural killer cells in vitro. When IL-18 antisense transfectants were implanted into syngeneic mice, severe reduction of tumor cell growth was observed with concomitant infiltrated natural killer cells in the tumor area. Taken together, these results demonstrate that IL-18 has a critical role as a survival factor for B16F10 melanoma cells. PMID- 10825145 TI - Cooperativity of Staphylococcal aureus enterotoxin B superantigen, major histocompatibility complex class II, and CD80 for immunotherapy of advanced spontaneous metastases in a clinically relevant postoperative mouse breast cancer model. AB - One of the leading causes of death for women is metastatic breast cancer. Because most animal tumors do not accurately model clinical metastatic disease, the development of effective therapies has progressed slowly. In this study, we establish the poorly immunogenic mouse 4T1 mammary carcinoma as a postsurgical animal model. 4T1 growth characteristics parallel highly invasive human metastatic mammary carcinoma and, at the time of surgery, the extent of disease is comparable with human stage IV breast cancer. Progress in understanding the immune response has led to innovative immune-based anticancer therapies. Here, we test in this postsurgical model, a novel cell-based vaccine, combining MHC class II, CD80(B7.1), and SEB superantigen. Effective treatment of tumor-bearing mice with this immunotherapy requires expression of all three molecules. Mean survival time is extended from 5-7.5 weeks for control-treated mice to 6-10.5 weeks for therapy-treated mice. Increased survival is accompanied by a maximum of 100-fold decrease in clonogenic lung metastases. These therapeutic effects are particularly noteworthy because: (a) the postoperative model demonstrates that early metastases responsible for morbidity are established by 2 weeks after tumor inoculation with 7 x 10(3) parental 4T1 cells into the mammary gland; (b) the immunotherapy is started 4 weeks after tumor inoculation when the mice contain extensive, pre-established, disseminated metastases; and (c) CD4+ and CD8+ T cells are required for the effect. PMID- 10825146 TI - Comparison of TP53 mutations identified by oligonucleotide microarray and conventional DNA sequence analysis. AB - As the rate of gene discovery accelerates, more efficient methods are needed to analyze genes in human tissues. To assess the efficiency, sensitivity, and specificity of different methods, alterations of TP53 were independently evaluated in 108 ovarian tumors by conventional DNA sequence analysis and oligonucleotide microarray (p53 GeneChip). All mutations identified by oligonucleotide microarray and all disagreements with conventional gel-based DNA sequence analysis were confirmed by re-analysis with manual and automated dideoxy DNA sequencing. A total of 77 ovarian cancers were identified as having TP53 mutations by one of the two approaches, 71 by microarray and 63 by gel-based DNA sequence analysis. The same mutation was identified in 57 ovarian cancers, and the same wild type TP53 sequence was observed in 31 ovarian cancers by both methods, for a concordance rate of 81%. Among the mutation analyses discordant by these methods for TP53 sequence were 14 cases identified as mutated by microarray but not by conventional DNA sequence analysis and 6 cases identified as mutated by conventional DNA sequence analysis but not by microarray. Overall, the oligonucleotide microarray demonstrated a 94% accuracy rate, a 92% sensitivity, and an 100% specificity. Conventional DNA sequence analysis demonstrated an 87% accuracy rate, 82% sensitivity, and a 100% specificity. Patients with TP53 mutations had significantly shorter overall survival than those with no mutation (P = 0.02). Women with mutations in loop2, loop3, or the loop-sheet-helix domain had shorter survival than women with other mutations or women with no mutations (P = 0.01). Although further refinement would be helpful to improve the detection of certain types of TPS3 alterations, oligonucleotide microarrays were shown to be a powerful and effective tool for TP53 mutation detection. PMID- 10825147 TI - Cytoplasmic sequestration and functional repression of p53 in the mammary epithelium is reversed by hormonal treatment. AB - Proper function of the p53 tumor suppressor gene is critical for inhibiting tumor development in a broad spectrum of tissues. Although the mammary gland is highly susceptible to tumor formation, the functional status of p53 in the normal tissue had not been investigated. Therefore, expression, localization, and activity of p53 were examined in normal mammary tissues. High levels of p53 protein were found expressed in the cytoplasm of the ductal epithelium of the quiescent mammary gland. Ionizing radiation failed to recruit p53 to the nucleus, and p53 dependent responses were minimal. However, transient hormonal stimulation resulted in nuclear accumulation of p53, an induction of p21/WAF1, and a 5-fold increase in apoptosis after ionizing radiation. Therefore, the functional state of wild-type p53 in the mammary epithelium can be regulated by hormonal stimuli. PMID- 10825148 TI - Small cell lung carcinoma exhibits greater phospholipase C-beta1 expression and edelfosine resistance compared with non-small cell lung carcinoma. AB - Aberrant signal transduction pathways involved in the development of metastatic disease are poorly defined in both small cell lung carcinoma (SCLC) and non-small cell lung carcinoma (NSCLC). Neuropeptide-driven positive feedback loops stimulating cell proliferation are characteristic of SCLC. The activation of phospholipase C (PLC)-beta1 is an early and common response to stimulation of G protein-coupled receptors by these neuroendocrine growth factors. The importance of PLC-beta in neuropeptide signaling prompted us to compare PLC-beta isoform expression and activity in four independent SCLC cell lines and four independent NSCLC cell lines. We found that PLC-beta1 is more highly expressed in SCLC than in NSCLC, as indicated by Western blotting of cell lysates. All SCLC lines studied express PLC-beta1; only one of the NSCLC lines investigated showed detectable levels of the enzyme. NSCLC lines are significantly more sensitive to the antiproliferative effects of ET-18-OCH3 (edelfosine) compared with the SCLC lines, as indicated by [3H]thymidine uptake. The only SCLC cell line (NCI-H345) that is as sensitive as the NSCLC cell lines to ET-18-OCH3 also expresses uniquely low levels of PLC-beta1. The participation of PLC-beta1 in signaling by SCLC growth factor receptors is indicated by our finding that PLC-beta1 (but not PLC-beta3) coimnunoprecipitates with G(alpha)q/11 upon activation of neurotensin receptors; this association is inhibited by ET-18-OCH3. Ca2+ mobilization mediated by neurotensin receptors is also inhibited by ET-18-OCH3. The binding of GTPgammaS to G(alpha)q/11 upon treatment of SCLC cells with neurotensin is not inhibited by ET-18-OCH3. These findings indicate that ET-18-OCH3 does not interfere with G(alpha)q/11 activation but rather inhibits the association of G(alpha)q/11 with PLC-beta1. Our data suggest that PLC-beta is an important mediator of both SCLC and NSCLC proliferation. Differences in PLC-beta1 expression may be exploitable in the development of effective diagnostic and therapeutic tools. PMID- 10825150 TI - Unique transcription pattern of Epstein-Barr virus (EBV) in EBV-carrying gastric adenocarcinomas: expression of the transforming BARF1 gene. AB - Approximately 10% of gastric adenocarcinomas worldwide are associated with human EBV. These carcinomas generally do not express the latent membrane protein 1 (LMP1), the major known EBV oncogene. Recently, another EBV gene [ie., BARF1 (BamHI A rightward open reading frame)] was shown to have transforming and immortalizing capacities. Therefore, in this study, we investigated the expression of BARF1 in EBV-carrying gastric adenocarcinomas in relation to the expression of other latent EBV transcripts. In the present study, 10 of 132 gastric adenocarcinomas tested positive for EBV using EBER1/2-RNA in situ hybridization. We demonstrate BARF1 gene transcription in nine EBV-carrying gastric adenocarcinomas (with sufficient RNA quality) using the BARF1-specific nucleic acid sequence-based amplification assay. In addition, we also detected other latent EBV transcripts (ie., BARF0-, LMP2A-, and Q/K-driven EBNA1 transcripts in these carcinomas using reverse transcription-PCR analysis. No expression of LMP1, EBNA2, and ZEBRA (either at transcription or protein level) was found. In addition, two cases were positive for BHRF1 transcripts, the viral bcl-2 homologue. Thus, together with BARF1 transcription, a unique and distinct EBV latency type has been found in EBV-associated gastric adenocarcinomas. Because BARF1 exerts immortalizing effects on human epithelial cells in vitro and EBV-carrying gastric adenocarcinomas lack the expression of LMP1, the BARF1 gene might act as the viral oncogene in EBV-carrying gastric carcinomas. The BARF1 gene offers an alternative way for EBV-mediated oncogenesis other than LMP1. PMID- 10825149 TI - Inducible pRb2/p130 expression and growth-suppressive mechanisms: evidence of a pRb2/p130, p27Kip1, and cyclin E negative feedback regulatory loop. AB - The retinoblastoma family of proteins, pRb/p105, p107, and pRb2/ p130, cooperate to regulate cell cycle progression through the G1 phase of the cell cycle. Each of the family members realize their common goal of G1-S checkpoint regulation through overlapping and unique growth regulatory pathways. We took advantage of a tetracycline-regulated gene expression system to control the expression of RB2/p130 in JC virus-induced hamster brain tumor cells to study in vivo the molecular mechanisms used by pRb2/p130 to elicit its growth-suppressive function. We have previously used this system to demonstrate that induction of pRb/ p130 expression suppresses tumor growth in vivo by overcoming neoplastic transformation mediated by the large T-antigen oncoprotein of JCV (JCV TAg). Here we found that induction of pRb2/p130 in vivo specifically inhibits cyclin A- and cyclin E-associated kinase activity and by doing so induces p27Kip1 levels presumably by inhibiting p27Kip1-targeted proteolysis by cyclin E-Cdk2 phosphorylation of p27Kip1. RB2/p130 induction also decreased cyclin A and the transcription factor E2F-1 while increasing cyclin E at both the transcriptional and protein levels of expression. The growth inhibitory activity of pRb2/p130 also correlated with its E2F-binding capacity. Furthermore, p27Kip1 and pRb2/p130 were found to be targets of the JCV TAg oncoprotein and to interact in vivo with each other independently from the presence of TAg. Interestingly, pRb2/p130 expression negatively modulated the binding of p27Kip1 to JCV TAg. These data suggest that pRb2/p130 and p27Kip1 may cooperate in regulating cellular proliferation, and both may be involved in a negative feedback regulatory loop with cyclin E. PMID- 10825151 TI - Identification of a tetrameric hedgehog signaling complex. AB - Hedgehog (Hh) signal transduction requires a large cytoplasmic multi-protein complex that binds microtubules in an Hh-dependent manner. Here, we show that three members of this complex, Costal2 (Cos2), Fused (Fu), and Cubitus interruptus (Ci), bind each other directly to form a trimeric complex. We demonstrate that this trimeric signaling complex exists in Drosophila lacking Suppressor of Fused (Su(fu)), an extragenic suppressor of fu, indicating that Su(fu) is not required for the formation, or apparently function, of the Hh signaling complex. However, we subsequently show that Su(fu), although not a requisite component of this complex, does form a tetrameric complex with Fu, Cos2, and Ci. This additional Su(fu)-containing Hh signaling complex does not appear to be enriched on microtubules. Additionally, we demonstrate that in response to Hh Ci accumulates in the nucleus without its various cytoplasmic binding partners, including Su(fu). We discuss a model in which Su(fu) and Cos2 each bind to Fu and Ci to exert some redundant effect on Ci such as cytoplasmic retention. This model is consistent with genetic data demonstrating that Su(fu) is not required for Hh signal transduction proper and with the elaborate genetic interactions observed among Su(fu), fu, cos2, and ci. PMID- 10825152 TI - Frequency-encoding Thr17 phospholamban phosphorylation is independent of Ser16 phosphorylation in cardiac myocytes. AB - Both Ser(16) and Thr(17) of phospholamban (PLB) are phosphorylated, respectively, by cAMP-dependent protein kinase (PKA) and Ca(2+)/calmodulin-dependent protein kinase II (CaMKII). PLB phosphorylation relieves cardiac sarcoplasmic reticulum Ca(2+) pump from inhibition by PLB. Previous studies have suggested that phosphorylation of Ser(16) by PKA is a prerequisite for Thr(17) phosphorylation by CaMKII and is essential to the relaxant effect of beta-adrenergic stimulation. To determine the role of Thr(17) PLB phosphorylation, we investigated the dual site phosphorylation of PLB in isolated adult rat cardiac myocytes in response to beta(1)-adrenergic stimulation or electrical field stimulation (0. 1-3 Hz) or both. A beta(1)-adrenergic agonist, norepinephrine (10(-9)-10(-6) m), in the presence of an alpha(1)-adrenergic antagonist, prazosin (10(-6) m), selectively increases the PKA-dependent phosphorylation of PLB at Ser(16) in quiescent myocytes. In contrast, electrical pacing induces an opposite phosphorylation pattern, selectively enhancing the CaMKII-mediated Thr(17) PLB phosphorylation in a frequency-dependent manner. When combined, electric stimulation (2 Hz) and beta(1)-adrenergic stimulation lead to dual phosphorylation of PLB and exert a synergistic effect on phosphorylation of Thr(17) but not Ser(16). Frequency dependent Thr(17) phosphorylation is closely correlated with a decrease in 50% relaxation time (t(50)) of cell contraction, which is independent of, but additive to, the relaxant effect of Ser(16) phosphorylation, resulting in hastened contractile relaxation at high stimulation frequencies. Thus, we conclude that in intact cardiac myocytes, phosphorylation of PLB at Thr(17) occurs in the absence of prior Ser(16) phosphorylation, and that frequencydependent Thr(17) PLB phosphorylation may provide an intrinsic mechanism for cardiac myocytes to adapt to a sudden change of heart rate. PMID- 10825153 TI - Calcium regulates transcriptional repression of myocyte enhancer factor 2 by histone deacetylase 4. AB - The myocyte enhancer factor 2 (MEF2) consists of a family of transcription factors that play important roles in a number of physiological processes from muscle cell differentiation to neuronal survival and T cell apoptosis. MEF2 has been reported to be associated with several distinct repressors including Cabin1(cain), MEF2-interacting transcriptional repressor (MITR), and HDAC4. It has been previously shown that Cabin1 is associated with MEF2 in a calcium sensitive manner; activated calmodulin binds to Cabin1 and releases it from MEF2. However, it was not known whether the binding of HDAC4 and MITR to MEF2 is also regulated by calcium. We report that HDAC4 and MITR contain calmodulin-binding domains that overlap with their MEF2-binding domains. Binding of calmodulin to HDAC4 leads to its dissociation from MEF2, relieving MEF2 from the transcriptional repression by HDAC4. Together, HDAC4, MITR, and Cabin1 constitute a family of calcium-sensitive transcriptional repressors of MEF2. PMID- 10825154 TI - Crystal structure determination of aristolochene synthase from the blue cheese mold, Penicillium roqueforti. AB - The 2.5-A resolution crystal structure of recombinant aristolochene synthase from the blue cheese mold, Penicillium roqueforti, is the first of a fungal terpenoid cyclase. The structure of the enzyme reveals active site features that participate in the cyclization of the universal sesquiterpene cyclase substrate, farnesyl diphosphate, to form the bicyclic hydrocarbon aristolochene. Metal triggered carbocation formation initiates the cyclization cascade, which proceeds through multiple complex intermediates to yield one exclusive structural and stereochemical isomer of aristolochene. Structural homology of this fungal cyclase with plant and bacterial terpenoid cyclases, despite minimal amino acid sequence identity, suggests divergence from a common, primordial ancestor in the evolution of terpene biosynthesis. PMID- 10825155 TI - Thermogenic responses in brown fat cells are fully UCP1-dependent. UCP2 or UCP3 do not substitute for UCP1 in adrenergically or fatty scid-induced thermogenesis. AB - To examine the thermogenic significance of the classical uncoupling protein-1 (UCP1), the thermogenic potential of brown adipocytes isolated from UCP1-ablated mice was investigated. Ucp1(-/-) cells had a basal metabolic rate identical to wild-type; the mitochondria within them were coupled to the same degree. The response to norepinephrine in wild-type cells was robust ( approximately 10-fold increase in thermogenesis); Ucp1(-/-) cells only responded approximately 3% of this. Ucp1(-/-) cells were as potent as wild-type in norepinephrine-induced cAMP accumulation and lipolysis and had a similar mitochondrial respiratory complement. In wild-type cells, fatty acids induced a thermogenic response similar to norepinephrine, but fatty acids (and retinoate) were practically without effect in Ucp1(-/-) cells. It is concluded that no other adrenergically induced thermogenic mechanism exists in brown adipocytes except that mediated by UCP1 and that entopic expression of UCP1 does not lead to overt innate uncoupling, and it is suggested that fatty acids are transformed to an intracellular physiological activator of UCP1. High expression of UCP2 and UCP3 in the tissue was not associated with an overt innate highly uncoupled state of mitochondria within the cells, nor with an ability of norepinephrine or endo- or exogenous fatty acids to induce uncoupled respiration in the cells. Thus, UCP1 remains the only physiologically potent thermogenic uncoupling protein in these cells. PMID- 10825156 TI - Overexpression of frequenin, a modulator of phosphatidylinositol 4-kinase, inhibits biosynthetic delivery of an apical protein in polarized madin-darby canine kidney cells. AB - Polyphosphoinositides regulate numerous steps in membrane transport. The levels of individual phosphatidylinositols are controlled by specific lipid kinases, whose activities and localization are in turn regulated by a variety of effectors. Here we have examined the effect of overexpression of frequenin, a modulator of phosphatidylinositol 4-kinase activity, on biosynthetic and postendocytic traffic in polarized Madin-Darby canine kidney cells. Endogenous frequenin was identified in these cells by polymerase chain reaction, Western blotting, and indirect immunofluorescence. Adenoviral-mediated overexpression of frequenin had no effect on early Golgi transport of membrane proteins, as assessed by acquisition of resistance to endoglycosidase H. However, delivery of newly synthesized influenza hemagglutinin from the trans-Golgi network to the apical cell surface was severely inhibited in cells overexpressing frequenin, whereas basolateral delivery of the polymeric immunoglobulin receptor was unaffected. Overexpression of frequenin did not affect postendocytic trafficking steps including apical and basolateral recycling and basal-to-apical transcytosis. We conclude that frequenin, and by inference, phosphatidylinositol 4-kinase, plays an important and selective role in apical delivery in polarized cells. PMID- 10825157 TI - Phosphorylation of c-Crk II on the negative regulatory Tyr222 mediates nerve growth factor-induced cell spreading and morphogenesis. AB - The Crk family of adaptor proteins participate in diverse signaling pathways that regulate growth factor-induced proliferation, anchorage-dependent DNA synthesis, and cytoskeletal reorganization, important for cell adhesion and motility. Using kidney epithelial 293T cells for transient co-transfection studies and the nerve growth factor (NGF)-responsive PC12 cell line as a model system for neuronal morphogenesis, we demonstrate that the non-receptor tyrosine kinase c-Abl is an intermediary for NGF-inducible c-Crk II phosphorylation on the negative regulatory Tyr(222). Transient expression of a c-Crk II Tyr(222) point mutant (c Crk Y222F) in 293T cells induces hyperphosphorylation of paxillin on Tyr(31) and enhances complex formation between c-Crk Y222F and paxillin as well as c-Crk Y222F and c-Abl, suggesting that c-Crk II Tyr(222) phosphorylation induces both the dissociation of the Crk SH2 domain from paxillin and the Crk SH3 domain from c-Abl. Interestingly, examination of the early kinetics of NGF stimulation in PC12 cells showed that c-Crk II Tyr(222) phosphorylation preceded paxillin Tyr(31) phosphorylation, followed by a transient initial dissociation of the c Crk II paxillin complex. PC12 cells overexpressing c-Crk Y222F manifested a defect in cellular adhesion and neuritogenesis that led to detachment of cells from the extracellular matrix, thus demonstrating the biological significance of c-Crk II tyrosine phosphorylation in NGF-dependent morphogenesis. Whereas previous studies have shown that Crk SH2 binding to paxillin is critical for cell adhesion and migration, our data show that the phosphorylation cycle of c-Crk II determines its dynamic interaction with paxillin, thereby regulating turnover of multiprotein complexes, a critical aspect of cytoskeletal plasticity and actin dynamics. PMID- 10825158 TI - Identification of residues of CXCR4 critical for human immunodeficiency virus coreceptor and chemokine receptor activities. AB - CXCR4 is a G-coupled receptor for the stromal cell-derived factor (SDF-1) chemokine, and a CD4-associated human immunodeficiency virus type 1 (HIV-1) coreceptor. These functions were studied in a panel of CXCR4 mutants bearing deletions in the NH(2)-terminal extracellular domain (NT) or substitutions in the NT, the extracellular loops (ECL), or the transmembrane domains (TMs). The coreceptor activity of CXCR4 was markedly impaired by mutations of two Tyr residues in NT (Y7A/Y12A) or at a single Asp residue in ECL2 (D193A), ECL3 (D262A), or TMII (D97N). These acidic residues could engage electrostatical interactions with basic residues of the HIV-1 envelope protein gp120, known to contribute to the selectivity for CXCR4. The ability of CXCR4 mutants to bind SDF 1 and mediate cell signal was consistent with the two-site model of chemokine receptor interaction. Site I involved in SDF-1 binding but not signaling was located in NT with particular importance of Glu(14) and/or Glu(15) and Tyr(21). Residues required for both SDF-1 binding and signaling, and thus probably part of site II, were identified in ECL2 (Asp(187)), TMII (Asp(97)), and TMVII (Glu(288)). The first residues () of NT also seem required for SDF-1 binding and signaling. A deletion in the third intracellular loop abolished signaling, probably by disrupting the coupling with G proteins. The identification of CXCR4 residues involved in the interaction with both SDF-1 and HIV-1 may account for the signaling activity of gp120 and has implications for the development of antiviral compounds. PMID- 10825159 TI - The Drosophila caspase DRONC cleaves following glutamate or aspartate and is regulated by DIAP1, HID, and GRIM. AB - The caspase family of cysteine proteases plays important roles in bringing about apoptotic cell death. All caspases studied to date cleave substrates COOH terminal to an aspartate. Here we show that the Drosophila caspase DRONC cleaves COOH-terminal to glutamate as well as aspartate. DRONC autoprocesses itself following a glutamate residue, but processes a second caspase, drICE, following an aspartate. DRONC prefers tetrapeptide substrates in which aliphatic amino acids are present at the P2 position, and the P1 residue can be either aspartate or glutamate. Expression of a dominant negative form of DRONC blocks cell death induced by the Drosophila cell death activators reaper, hid, and grim, and DRONC overexpression in flies promotes cell death. Furthermore, the Drosophila cell death inhibitor DIAP1 inhibits DRONC activity in yeast, and DIAP1's ability to inhibit DRONC-dependent yeast cell death is suppressed by HID and GRIM. These observations suggest that DRONC acts to promote cell death. However, DRONC activity is not suppressed by the caspase inhibitor and cell death suppressor baculovirus p35. We discuss possible models for DRONC function as a cell death inhibitor. PMID- 10825160 TI - Functional cloning and characterization of a novel nonhomeodomain protein that inhibits the binding of PBX1-HOX complexes to DNA. AB - PBX1 is a homeodomain protein that functions in complexes with other homeodomain containing proteins to regulate gene expression during developmental and/or differentiation processes. A yeast two-hybrid screen of a fetal liver hematopoietic cDNA library using PBX1a as bait led to the discovery of a novel non-homeodomain-containing protein that interacts with PBX1 as well as PBX2 and PBX3. RNA analysis revealed it to be expressed in CD34(+) hematopoietic cell populations enriched in primitive progenitors, as is PBX1; search of the expressed sequence tag data base indicated that it is also expressed in other early embryonic as well as adult tissues. The full-length cDNA encodes a 731 amino acid protein that has no significant homology to known proteins. This protein that we have termed hematopoietic PBX-interacting protein (HPIP) is mainly localized in the cytosol and in small amounts in the nucleus. The region of PBX that interacts with HPIP includes both the homeodomain and immediate N terminal flanking sequences. Strikingly, electrophoretic mobility shift assays revealed that HPIP inhibits the ability of PBX-HOX heterodimers to bind to target sequences. Moreover, HPIP strongly inhibits the transactivation activity of E2A PBX. Together these findings suggest that HPIP is a new regulator of PBX function. PMID- 10825161 TI - Identification of a 30-base pair regulatory element and novel DNA binding protein that regulates the human GLUT4 promoter in transgenic mice. AB - We have previously demonstrated that the important cis-acting elements regulating transcription of the human GLUT4 gene reside within 895 base pairs (bp) upstream of the transcription initiation site (Thai, M. V., Guruswamy, S., Cao, K. T., Pessin, J. E., and Olson, A. L. (1998) J. Biol. Chem. 273, 14285-14292). Our studies demonstrated that an MEF2 binding site within this region was necessary, but not sufficient, for GLUT4 promoter function in transgenic mice. We have identified a second regulatory element (Domain I) that functions cooperatively with the MEF2 domain in regulating GLUT4 transcription. Using a yeast-one hybrid screen, we obtained a partial cDNA and generated an antibody directed against a protein binding specifically to Domain I. Sequence analysis of the partial cDNA indicates that the protein binding to Domain I is a novel protein. The antibody specifically labels two proteins of approximately 70 and 50 kDa in Western blot analysis. These molecular masses correspond to Domain I binding proteins identified by UV-cross-linking nuclear extracts to a Domain I probe. The antibody raised against the Domain I binding protein inhibited formation of a Domain I protein complex in electrophoretic mobility shift assays. We conclude that we have identified an authentic, novel, Domain I binding protein required for transcriptional regulation of the human GLUT4 promoter. PMID- 10825162 TI - Replication protein A physically interacts with the Bloom's syndrome protein and stimulates its helicase activity. AB - Bloom's syndrome is a rare autosomal recessive disorder characterized by genomic instability and predisposition to cancer. BLM, the gene defective in Bloom's syndrome, encodes a 159-kDa protein possessing DNA-stimulated ATPase and ATP dependent DNA helicase activities. We have examined mechanistic aspects of the catalytic functions of purified recombinant BLM protein. Through analyzing the effects of different lengths of DNA cofactor on ATPase activity, we provide evidence to suggest that BLM translocates along single-stranded DNA in a processive manner. The helicase reaction catalyzed by BLM protein was examined as a function of duplex DNA length. We show that BLM catalyzes unwinding of short DNA duplexes (/=259-bp). The presence of the human single-stranded DNA-binding protein (human replication protein A (hRPA)) stimulates the BLM unwinding reaction on the 259-bp partial duplex DNA substrate. Heterologous single-stranded DNA-binding proteins fail to stimulate similarly the helicase activity of BLM protein. This is the first demonstration of a functional interaction between BLM and another protein. Consistent with a functional interaction between hRPA and the BLM helicase, we demonstrate a direct physical interaction between the two proteins mediated by the 70-kDa subunit of RPA. The interactions between BLM and hRPA suggest that the two proteins function together in vivo to unwind DNA duplexes during replication, recombination, or repair. PMID- 10825163 TI - p75 reduces TrkB tyrosine autophosphorylation in response to brain-derived neurotrophic factor and neurotrophin 4/5. AB - Neurotrophins mediate their signals through two different receptors: the family of receptor tyrosine kinases, Trks, and the low affinity pan-neurotrophin receptor p75. Trk receptors show more restricted ligand specificity, whereas all neurotrophins are able to bind to p75. One important function of p75 is the enhancement of nerve growth factor signaling via TrkA by increasing TrkA tyrosine autophosphorylation. Here, we have examined the importance of p75 on TrkB- and TrkC-mediated neurotrophin signaling in an MG87 fibroblast cell line stably transfected with either p75 and TrkB or p75 and TrkC, as well as in PC12 cells stably transfected with TrkB. In contrast to TrkA signaling, p75 had a negative effect on TrkB tyrosine autophosphorylation in response to its cognate neurotrophins, brain-derived neurotrophic factor and neurotrophin 4/5. On the other hand, p75 had no effect on TrkB or TrkC activation in neurotrophin 3 treatment. p75 did not effect extracellular signal-regulated kinase 2 tyrosine phosphorylation in response to brain-derived neurotrophic factor, neurotrophin 3, or neurotrophin 4/5. These results suggest that the observed reduction in TrkB tyrosine autophosphorylation caused by p75 does not influence Ras/mitogen activated protein kinase signaling pathway in neurotrophin treatments. PMID- 10825164 TI - HPC3 is a new human polycomb orthologue that interacts and associates with RING1 and Bmi1 and has transcriptional repression properties. AB - Polycomb group (PcG) proteins were first described in Drosophila as factors responsible for maintaining the transcriptionally repressed state of Hox/homeotic genes in a stable and heritable manner throughout development. A growing number of vertebrate genes related to the Drosophila PcG proteins have recently been identified, including two Polycomb orthologues, Pc2 and M33. PcG proteins form multiprotein complexes, termed PcG bodies, that are thought to repress transcription by altering chromatin structure. Here we report the identification and characterization of HPC3 (human Polycomb 3), a novel PcG protein isolated in a yeast two-hybrid screen using human RING1 as bait. HPC3 shows strong sequence similarity to Drosophila Pc and also to vertebrate Pc2 and M33, particularly within the chromodomain and C-box. Previous studies indicate that M33 and human Pc2 (HPC2) can interact with RING1, and we show here that HPC3 also binds to RING1. This interaction is dependent upon the HPC3 C-box but, only partially on the RING finger of RING1. In contrast to HPC2, HPC3 interactions with RING1 are only observed in vivo with covalently modified forms of RING1. HPC3 also colocalizes with other PcG proteins in human PcG bodies. Consistent with its role as a PcG member, HPC3 is able to act as a long range transcriptional silencer when targeted to a reporter gene by a heterologous DNA-binding domain. Taken together, these data suggest that HPC3 is part of a large multiprotein complex that also contains other PcG proteins and is involved in repression of transcriptional activity. PMID- 10825165 TI - Conditional expression of RNA polymerase II in mammalian cells. Deletion of the carboxyl-terminal domain of the large subunit affects early steps in transcription. AB - The carboxyl-terminal domain (CTD) of the large subunit of mammalian RNA polymerase II contains 52 repeats of a heptapeptide that is the target of a variety of kinases. The hyperphosphorylated CTD recruits important factors for mRNA capping, splicing, and 3'-processing. The role of the CTD for the transcription process in vivo, however, is not yet clear. We have conditionally expressed an alpha-amanitin-resistant large subunit with an almost entirely deleted CTD (LS*Delta5) in B-cells. These cells have a defect in global transcription of cellular genes in the presence of alpha-amanitin. Moreover, pol II harboring LS*Delta5 failed to transcribe up to the promoter-proximal pause sites in the hsp70A and c-fos gene promoters. The results indicate that the CTD is already required for steps that occur before promoter-proximal pausing and maturation of mRNA. PMID- 10825166 TI - Identification of a novel ligand-receptor pair constitutively activated by ras oncogenes. AB - The Ras signaling pathway is thought to control the expression of a subset of yet to be defined genes that are crucial for cell growth and differentiation. Here we have identified by differential display a novel oncogenic Ras target, mob-5, encoding a 23-kDa cytokine-like secreted protein. Mob-5 expression could be induced by oncogenic Ha-ras and Ki-ras, but not by normal ras activation. Inhibitors of both Ha-Ras and mitogen-activated protein kinase kinase completely abolished the mob-5 expression in ras transformed cells, with concomitant loss of the transformation phenotype. Using an alkaline phosphatase-tagged Mob-5 as ligand, a putative Mob-5 receptor was identified on the cell surface of oncogenic ras transformed cells. Thus, the Mob-5/Mob-5 receptor may represent a novel putative autocrine loop coordinately activated by ras oncogenes. PMID- 10825167 TI - Role of the intracellular domain of the human type I interferon receptor 2 chain (IFNAR2c) in interferon signaling. Expression of IFNAR2c truncation mutants in U5A cells. AB - A human cell line (U5A) lacking the type I interferon (IFN) receptor chain 2 (IFNAR2c) was used to determine the role of the IFNAR2c cytoplasmic domain in regulating IFN-dependent STAT activation, interferon-stimulated gene factor 3 (ISGF3) and c-sis-inducible factor (SIF) complex formation, gene expression, and antiproliferative effects. A panel of U5A cells expressing truncation mutants of IFNAR2c on their cell surface were generated for study. Janus kinase (JAK) activation was detected in all mutant cell lines; however, STAT1 and STAT2 activation was observed only in U5A cells expressing full-length IFNAR2c and IFNAR2c truncated at residue 462 (R2.462). IFNAR2c mutants truncated at residues 417 (R2. 417) and 346 (R2.346) or IFNAR2c mutant lacking tyrosine residues in its cytoplasmic domain (R2.Y-F) render the receptor inactive. A similar pattern was observed for IFN-inducible STAT activation, STAT complex formation, and STAT-DNA binding. Consistent with these data, IFN-inducible gene expression was ablated in U5A, R2.Y-F, R2.417, and R2.346 cell lines. The implications are that tyrosine phosphorylation and the 462-417 region of IFNAR2c are independently obligatory for receptor activation. In addition, the distal 53 amino acids of the intracellular domain of IFNAR2c are not required for IFN-receptor mediated STAT activation, ISFG3 or SIF complex formation, induction of gene expression, and inhibition of thymidine incorporation. These data demonstrate for the first time that both tyrosine phosphorylation and a specific domain of IFNAR2c are required in human cells for IFN-dependent coupling of JAK activation to STAT phosphorylation, gene induction, and antiproliferative effects. In addition, human and murine cells appear to require different regions of the cytoplasmic domain of IFNAR2c for regulation of IFN responses. PMID- 10825168 TI - Site-directed mutagenesis of the yeast resolving enzyme Cce1 reveals catalytic residues and relationship with the intron-splicing factor Mrs1. AB - The Holliday junction-resolving enzyme Cce1 is a magnesium-dependent endonuclease, responsible for the resolution of recombining mitochondrial DNA molecules in Saccharomyces cerevisiae. We have identified a homologue of Cce1 from Candida albicans and used a multiple sequence alignment to predict residues important for junction binding and catalysis. Twelve site-directed mutants have been constructed, expressed, purified, and characterized. Using this approach, we have identified basic residues with putative roles in both DNA recognition and catalysis of strand scission and acidic residues that have a purely catalytic role. We have shown directly by isothermal titration calorimetry that a group of acidic residues vital for catalytic activity in Cce1 act as ligands for the catalytic magnesium ions. Sequence similarities between the Cce1 proteins and the group I intron splicing factor Mrs1 suggest the latter may also possess a binding site for magnesium, with a putative role in stabilization of RNA tertiary structure or catalysis of the splicing reaction. PMID- 10825169 TI - Transforming growth factor-beta 1 inhibits cytokine-mediated induction of human metalloelastase in macrophages. AB - Matrix metalloproteinases (MMP) have been identified in vulnerable areas of atherosclerotic plaques and may contribute to plaque instability through extracellular matrix degradation. Human metalloelastase (MMP-12) is a macrophage specific MMP with broad substrate specificity and is capable of degrading proteins found in the extracellular matrix of atheromas. Despite its potential importance, little is known about the regulation of MMP-12 expression in the context of atherosclerosis. In this study, we report that in human peripheral blood-derived macrophages, MMP-12 mRNA was markedly up-regulated by several pro atherosclerotic cytokines and growth factors including interleukin-1beta, tumor necrosis factor-alpha, macrophage colony-stimulating factor, vascular endothelial growth factor, and platelet-derived growth factor-BB. In contrast, the pleiotropic anti-inflammatory growth factor transforming growth factor-beta1 (TGF beta1) inhibited cytokine-mediated induction of MMP-12 mRNA, protein, and enzymatic activity. Analyses of MMP-12 promoter through transient transfections and electrophoretic mobility shift assays indicated that both its induction by cytokines and its inhibition by TGF-beta1 depended on signaling through an AP-1 site at -81 base pairs. Moreover, the inhibitory effect of TGF-beta1 on MMP-12 was dependent on Smad3. Taken together, MMP-12 is induced by several factors implicated in atherosclerosis. The inhibition of MMP-12 expression by TGF-beta1 suggests that TGF-beta1, acting via Smad3, may promote plaque stability. PMID- 10825170 TI - The C-terminal alpha helix of Tn5 transposase is required for synaptic complex formation. AB - An important step in Tn5 transposition requires transposase-transposase homodimerization to form a synaptic complex competent for cleavage of transposon DNA free from the flanking sequence. We demonstrate that the C-terminal helix of Tn5 transposase (residues 458-468 of 476 total amino acids) is required for synaptic complex formation during Tn5 transposition. Specifically, deletion of eight amino acids or more from the C terminus greatly reduces or abolishes synaptic complex formation in vitro. Due to this impaired synaptic complex formation, transposases lacking eight amino acids are also defective in the cleavage step of transposition. Interactions within the synaptic complex dimer interface were investigated by site-directed mutagenesis, and residues required for synaptic complex formation include amino acids comprising the dimer interface in the Tn5 inhibitor x-ray crystal structure dimer. Because the crystal structure dimer was hypothesized to be the inhibitory complex and not a synaptic complex, this result was surprising. Based on these data, models for both in vivo and in vitro synaptic complex formation are presented. PMID- 10825171 TI - Inhibition of toxicity in the beta-amyloid peptide fragment beta -(25-35) using N methylated derivatives: a general strategy to prevent amyloid formation. AB - beta-(25-35) is a synthetic derivative of beta-amyloid, the peptide that is believed to cause Alzheimer's disease. As it is highly toxic and forms fibrillar aggregates typical of beta-amyloid, it is suitable as a model for testing inhibitors of aggregation and toxicity. We demonstrate that N-methylated derivatives of beta-(25-35), which in isolation are soluble and non-toxic, can prevent the aggregation and inhibit the resulting toxicity of the wild type peptide. N-Methylation can block hydrogen bonding on the outer edge of the assembling amyloid. The peptides are assayed by Congo red and thioflavin T binding, electron microscopy, and a 3-(4, 5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) toxicity assay on PC12 cells. One peptide (Gly(25) N-methylated) has properties similar to the wild type, whereas five have varying effects on prefolded fibrils and fibril assembly. In particular, beta-(25 35) with Gly(33) N-methylated is able to completely prevent fibril assembly and to reduce the toxicity of prefolded amyloid. With Leu(34) N-methylated, the fibril morphology is altered and the toxicity reduced. We suggest that the use of N-methylated derivatives of amyloidogenic peptides and proteins could provide a general solution to the problem of amyloid deposition and toxicity. PMID- 10825172 TI - Analysis by NMR spectroscopy of the structural homology between the linear and the cyclic peptide recognized by anti-human leukocyte antigen class I monoclonal antibody TP25.99*. AB - The anti-human leukocyte antigen (HLA) class I monoclonal antibody (mAb) TP25.99 has a unique specificity since it recognizes both a conformational and a linear determinant expressed on the beta(2)-mu-associated and beta(2)-mu-free HLA class I heavy chains, respectively. Previously, we reported the identification of a cyclic and a linear peptide that inhibits mAb TP25.99 binding to the beta(2)-mu associated and beta(2)-mu-free HLA class I heavy chains (S. A. Desai, X. Wang, E. J. Noronha, Q. Zhou, V. Rebmann, H. Grosse-Wilde, F. J. Moy, R. Powers, and S. Ferrone, submitted for publication). The linear X(19) and cyclic LX-8 peptides contain sequence homologous to residues 239-242, 245, and 246 and to residues 194 198, respectively, of HLA class I heavy chain alpha(3) domain. Analysis by two dimensional transfer nuclear Overhauser effect spectroscopy of the induced solution structures of the linear X(19) and cyclic LX-8 peptides in the presence of mAb TP25.99 showed that the two peptides adopt a similar structural motif despite the lack of sequence homology. The backbone fold is suggestive of a short helical segment followed by a tight turn, reminiscent of the determinant loop region (residues 194-198) on beta(2)-mu-associated HLA class I heavy chains. The structural similarity between the linear X(19) and cyclic LX-8 peptides and the lack of sequence homology suggests that mAb TP25.99 predominantly recognizes a structural motif instead of a consensus sequence. PMID- 10825173 TI - Interaction of tropoelastin with the amino-terminal domains of fibrillin-1 and fibrillin-2 suggests a role for the fibrillins in elastic fiber assembly. AB - Alignment of tropoelastin molecules during the process of elastogenesis is thought to require fibrillin-containing microfibrils. In this study, we have demonstrated that amino-terminal domains of two microfibrillar proteins, fibrillin-1 and fibrillin-2, interact with tropoelastin in solid phase binding assays. The tropoelastin-binding site was localized to a region beginning at the glycine-rich and proline-rich regions of fibrillin-2 and fibrillin-1, respectively, and continuing through the second 8-cysteine domain. Characterization of the binding requirements using the fibrillin-2 construct found that a folded, secondary structure was necessary for binding. Furthermore, binding between tropoelastin and fibrillin was mediated by ionic interactions involving the lysine side chains of tropoelastin. The importance of the lysine side chains was corroborated by the finding that the fibrillin-2 construct did not bind to mature elastin, whose lysine side chains have been modified to form cross-links. Interestingly, there was no interaction between the fibrillin constructs and tropoelastin in solution phase, suggesting that binding of tropoelastin to a solid substrate exposes a cryptic binding site. These results suggest that fibrillin plays an important role in elastic fiber assembly by binding tropoelastin and perhaps facilitating side chain alignment for efficient cross-linking. PMID- 10825174 TI - The yeast histone acetyltransferase A2 complex, but not free Gcn5p, binds stably to nucleosomal arrays. AB - We have investigated the structural basis for the differential catalytic function of the yeast Gcn5p-containing histone acetyltransferase (HAT) A2 complex and free recombinant yeast Gcn5p (rGcn5p). HAT A2 is shown to be a unique complex that contains Gcn5p, Ada2p, and Ada3p, but not proteins specific to other related HAT A complexes, e.g. ADA, SAGA. Nevertheless, HAT A2 produces the same unique polyacetylation pattern of nucleosomal substrates reported previously for ADA and SAGA, demonstrating that proteins specific to the ADA and SAGA complexes do not influence the enzymatic activity of Gcn5p within the HAT A2 complex. To investigate the role of substrate interactions in the differential behavior of free and complexed Gcn5p, sucrose density gradient centrifugation was used to characterize the binding of HAT A2 and free rGcn5p to intact and trypsinized nucleosomal arrays, H3/H4 tetramer arrays, and nucleosome core particles. We find that HAT A2 forms stable complexes with all nucleosomal substrates tested. In distinct contrast, rGcn5p does not interact stably with nucleosomal arrays, despite being able to specifically monoacetylate the H3 N terminus of nucleosomal substrates. Our data suggest that the ability of the HAT A2 complex to bind stably to nucleosomal arrays is functionally related to both local and global acetylation by the complexed and free forms of Gcn5p. PMID- 10825175 TI - Mechanism of kappa B DNA binding by Rel/NF-kappa B dimers. AB - The DNA binding of three different NF-kappaB dimers, the p50 and p65 homodimers and the p50/p65 heterodimer, has been examined using a combination of gel mobility shift and fluorescence anisotropy assays. The NF-kappaB p50/p65 heterodimer is shown here to bind the kappaB DNA target site of the immunoglobulin kappa enhancer (Ig-kappaB) with an affinity of approximately 10 nm. The p50 and p65 homodimers bind to the same site with roughly 5- and 15-fold lower affinity, respectively. The nature of the binding isotherms indicates a cooperative mode of binding for all three dimers to the DNA targets. We have further characterized the role of pH, salt, and temperature on the formation of the p50/p65 heterodimer-Ig-kappaB complex. The heterodimer binds to the Ig-kappaB DNA target in a pH-dependent manner, with the highest affinity between pH 7.0 and 7.5. A strong salt-dependent interaction between Ig-kappaB and the p50/p65 heterodimer is observed, with optimum binding occurring at monovalent salt concentrations below 75 mm, with binding becoming virtually nonspecific at a salt concentration of 200 mm. Binding of the heterodimer to DNA was unchanged across a temperature range between 4 degrees C and 42 degrees C. The sensitivity to ionic environment and insensitivity to temperature indicate that NF-kappaB p50/p65 heterodimers form complexes with specific DNA in an entropically driven manner. PMID- 10825176 TI - The high resolution crystal structure for class A beta-lactamase PER-1 reveals the bases for its increase in breadth of activity. AB - The treatment of infectious diseases by beta-lactam antibiotics is continuously challenged by the emergence and dissemination of new beta-lactamases. In most cases, the cephalosporinase activity of class A enzymes results from a few mutations in the TEM and SHV penicillinases. The PER-1 beta-lactamase was characterized as a class A enzyme displaying a cephalosporinase activity. This activity was, however, insensitive to the mutations of residues known to be critical for providing extended substrate profiles to TEM and SHV. The x-ray structure of the protein, solved at 1.9-A resolution, reveals that two of the most conserved features in class A beta-lactamases are not present in this enzyme: the fold of the Omega-loop and the cis conformation of the peptide bond between residues 166 and 167. The new fold of the Omega-loop and the insertion of four residues at the edge of strand S3 generate a broad cavity that may easily accommodate the bulky substituents of cephalosporin substrates. The trans conformation of the 166-167 bond is related to the presence of an aspartic acid at position 136. Selection of class A enzymes based on the occurrence of both Asp(136) and Asn(179) identifies a subgroup of enzymes with high sequence homology. PMID- 10825177 TI - Expression and function of the mouse collagen receptor glycoprotein VI is strictly dependent on its association with the FcRgamma chain. AB - Platelet glycoprotein (GP) VI has been proposed as the major collagen receptor for activation of human platelets. Human GPVI belongs to the immunoglobulin superfamily and is noncovalently associated with the FcRgamma chain that is involved in signaling through the receptor. In mice, similar mechanisms seem to exist as platelets from FcRgamma chain-deficient mice do not aggregate in response to collagen. However, the activating collagen receptor on mouse platelets has not been definitively identified. In the current study we examined the function and in vivo expression of GPVI in control and FcRgamma chain deficient mice with the first monoclonal antibody against GPVI (JAQ1). On wild type platelets, JAQ1 inhibited platelet aggregation induced by collagen but not PMA or thrombin. Cross-linking of bound JAQ1, on the other hand, induced aggregation of wild type but not FcRgamma chain-deficient platelets. JAQ1 stained platelets and megakaryocytes from wild type but not FcRgamma chain-deficient mice. Furthermore, JAQ1 recognized GPVI (approximately 60 kDa) in immunoprecipitation and Western blot experiments with wild type but not FcRgamma chain-deficient platelets. These results strongly suggest that GPVI is the collagen receptor responsible for platelet activation in mice and demonstrate that the association with the FcRgamma chain is critical for its expression and function. PMID- 10825178 TI - Sp1 increases expression of cyclooxygenase-2 in hypoxic vascular endothelium. Implications for the mechanisms of aortic aneurysm and heart failure. AB - Cyclooxygenase-2 (COX-2) catalyzes prostaglandin synthesis from arachidonic acid and is expressed locally in aortic aneurysm and heart failure. Cellular hypoxia is also found in these conditions. We have previously shown that cox-2 is transcriptionally regulated by hypoxia in human umbilical vein endothelial cells (HUVEC) in culture via the transactivation factor NF-kappaB p65, leading to increased production of prostaglandin E(2), an inhibitor of vascular smooth muscle cell proliferation. Sp1 is a transactivation factor known to be important in the regulation of cytokine expression in association with NF-kappaB. We hypothesized that Sp1 is involved in the induction of cox-2 in hypoxic HUVEC. Electrophoretic mobility shift assays with hypoxic HUVEC nuclear protein showed that both Sp1 and the related protein Sp3 specifically bound to the cox-2 promoter. Immunoblotting demonstrated that hypoxia increased the nuclear localization of Sp1 but did not change the Sp3 content in HUVEC. Overexpression of Sp1 through transfection of HUVEC enhanced cox-2 promoter activity as measured by reporter gene expression and by the production of COX-2. The specificity of the results was confirmed by mutation of the Sp1-binding site in the cox-2 promoter construct and by reproducibility in an Sp-deficient Drosophila SL2 cell line. The regulatory role of Sp1 discovered in this work supports the concept that a mechanistic link exists between vascular cellular hypoxia and mediators of inflammation associated with aortic aneurysm and heart failure. PMID- 10825179 TI - Selective methylation changes on the Bacillus subtilis chemotaxis receptor McpB promote adaptation. AB - The Bacillus subtilis McpB is a class III chemotaxis receptor, from which methanol is released in response to all stimuli. McpB has four putative methylation sites based upon the Escherichia coli consensus sequence. To explore the nature of methanol release from a class III receptor, all combinations of putative methylation sites Gln(371), Gln(595), Glu(630), and Glu(637) were substituted with aspartate, a conservative substitution that effectively eliminates methylation. McpB((Q371D,E630D,E637D)) in a Delta(mcpA mcpB tlpA tlpB)101::cat mcpC4::erm background failed to release methanol in response to either the addition or removal of the McpB-mediated attractant asparagine. In the same background, McpB((E630D,E637D)) produced methanol only upon asparagine addition, whereas McpB((Q371D,E630D)) produced methanol only upon asparagine removal. Thus methanol release from McpB was selective. Mutants unable to methylate site 637 but able to methylate site 630 had high prestimulus biases and were incapable of adapting to asparagine addition. Mutants unable to methylate site 630 but able to methylate site 637 had low prestimulus biases and were impaired in adaptation to asparagine removal. We propose that selective methylation of these two sites represents a method of adaptation novel from E. coli and present a model in which a charged residue rests between them. The placement of this charge would allow for opposing electrostatic effects (and hence opposing receptor conformational changes). We propose that CheC, a protein not found in enteric systems, has a role in regulating this selective methylation. PMID- 10825180 TI - Molecular characterization of the yeast vacuolar H+-ATPase proton pore. AB - The Saccharomyces cerevisiae vacuolar ATPase (V-ATPase) is composed of at least 13 polypeptides organized into two distinct domains, V(1) and V(0), that are structurally and mechanistically similar to the F(1)-F(0) domains of the F-type ATP synthases. The peripheral V(1) domain is responsible for ATP hydrolysis and is coupled to the mechanism of proton translocation. The integral V(0) domain is responsible for the translocation of protons across the membrane and is composed of five different polypeptides. Unlike the F(0) domain of the F-type ATP synthase, which contains 12 copies of a single 8-kDa proteolipid, the V-ATPase V(0) domain contains three proteolipid species, Vma3p, Vma11p, and Vma16p, with each proteolipid contributing to the mechanism of proton translocation (Hirata, R., Graham, L. A., Takatsuki, A., Stevens, T. H., and Anraku, Y. (1997) J. Biol. Chem. 272, 4795-4803). Experiments with hemagglutinin- and c-Myc epitope-tagged copies of the proteolipids revealed that each V(0) complex contains all three species of proteolipid with only one copy each of Vma11p and Vma16p but multiple copies of Vma3p. Since the proteolipids of the V(0) complex are predicted to possess four membrane-spanning alpha-helices, twice as many as a single F-ATPase proteolipid subunit, only six V-ATPase proteolipids would be required to form a hexameric ring-like structure similar to the F(0) domain. Therefore, each V(0) complex will likely be composed of four copies of the Vma3p proteolipid in addition to Vma11p and Vma16p. Structural differences within the membrane spanning domains of both V(0) and F(0) may account for the unique properties of the ATP-hydrolyzing V-ATPase compared with the ATP-generating F-type ATP synthase. PMID- 10825181 TI - c-Jun N-terminal kinase is essential for growth of human T98G glioblastoma cells. AB - The c-Jun N-terminal kinase/stress-activated protein kinase (JNK/SAPK) pathway is activated by numerous cellular stresses. Although it has been implicated in mediating apoptosis and growth factor signaling, its role in regulating cell growth is not yet clear. Here, the influence of JNK on basal (unstimulated) growth of human tumor glioblastoma T98G cells was investigated using highly specific JNK antisense oligonucleotides to inhibit JNK expression. Transient depletion of either JNK1 or JNK2 suppressed cell growth associated with an inhibition of DNA synthesis and cell cycle arrest in S phase. The growth inhibitory potency of JNK2 antisense ((JNK)2 IC(50) = 0.14 micrometer) was greater than that of JNK1 antisense ((JNK)1 IC(50) = 0.37 micrometer), suggesting that JNK2 plays a dominant role in regulating growth of T98G cells. Indeed, JNK2 antisense-treated populations exhibited greater inhibition of DNA synthesis and accumulation of S-phase cells than did the JNK1 antisense-treated cultures, with a significant proportion of these cells detaching from the tissue culture plate. JNK2 (but not JNK1) antisense-treated cultures exhibited marked elevation in the expression of the cyclin-dependent kinase inhibitor p21(cip1/waf1) accompanied by inhibition of Cdk2/Cdc2 kinase activities. Taken together, these results indicate that JNK is required for growth of T98G cells in nonstress conditions and that p21(cip1/waf1) may contribute to the sustained growth arrest of JNK2-depleted T98G cultures. PMID- 10825182 TI - Association of the N- and C-terminal domains of phospholipase D. Contribution of the conserved HKD motifs to the interaction and the requirement of the association for Ser/Thr phosphorylation of the enzyme. AB - Rat brain phospholipase D1 (rPLD1) belongs to a superfamily defined by the highly conserved catalytic motif (H(X)K(X)(4)D, denoted HKD. rPLD1 contains two HKD domains, located in the N- and C-terminal regions. The integrity of the two HKD domains is essential for enzymatic activity. Our previous studies showed that the N-terminal half of rPLD1 containing one HKD motif can associate with the C terminal half containing the other HKD domain to reconstruct wild type PLD activity (Xie, Z., Ho, W.-T. and Exton, J. H. (1998) J. Biol. Chem. 273, 34679 34682). In the present study, we have shown by mutagenesis that conserved amino acids in the HKD domains are important for both the catalytic activity and the association between the two halves of rPLD1. Furthermore, we found that rPLD1 could be modified by Ser/Thr phosphorylation. The modification occurred at the N terminal half of the enzyme, however, the association of the N-terminal domain with the C-terminal domain was required for the modification. The phosphorylation of the enzyme was not required for its catalytic activity or response to PKCalpha and small G proteins in vitro, although the phosphorylated form of rPLD1 was localized exclusively in the crude membrane fraction. In addition, we found that the individually expressed N- and C-terminal fragments did not interact when mixed in vitro and were unable to reconstruct PLD activity under these conditions. It is concluded that the association of the N- and C-terminal halves of rPLD1 requires their co-expression in vivo and depends on conserved residues in the HKD domains. The association is also required for Ser/Thr phosphorylation of the enzyme. PMID- 10825183 TI - Nuclear RNA export pathways. PMID- 10825184 TI - Stem-loop binding protein, the protein that binds the 3' end of histone mRNA, is cell cycle regulated by both translational and posttranslational mechanisms. AB - The expression of the replication-dependent histone mRNAs is tightly regulated during the cell cycle. As cells progress from G(1) to S phase, histone mRNA levels increase 35-fold, and they decrease again during G(2) phase. Replication dependent histone mRNAs are the only metazoan mRNAs that lack polyadenylated tails, ending instead in a conserved stem-loop. Much of the cell cycle regulation is posttranscriptional and is mediated by the 3' stem-loop. A 31-kDa stem-loop binding protein (SLBP) binds the 3' end of histone mRNA. The SLBP is necessary for pre-mRNA processing and accompanies the histone mRNA to the cytoplasm, where it is a component of the histone messenger RNP. We used synchronous CHO cells selected by mitotic shakeoff and HeLa cells synchronized at the G(1)/S or the M/G(1) boundary to study the regulation of SLBP during the cell cycle. In each system the amount of SLBP is regulated during the cell cycle, increasing 10- to 20-fold in the late G(1) and then decreasing in the S/G(2) border. SLBP mRNA levels are constant during the cell cycle. SLBP is regulated at the level of translation as cells progress from G(1) to S phase, and the protein is rapidly degraded as they progress into G(2). Regulation of SLBP may account for the posttranscriptional component of the cell cycle regulation of histone mRNA. PMID- 10825185 TI - Two regulators of Ste12p inhibit pheromone-responsive transcription by separate mechanisms. AB - The yeast Saccharomyces cerevisiae transcription factor Ste12p is responsible for activating genes in response to MAP kinase cascades controlling mating and filamentous growth. Ste12p is negatively regulated by two inhibitor proteins, Dig1p (also called Rst1p) and Dig2p (also called Rst2p). The expression of a C terminal Ste12p fragment (residues 216 to 688) [Ste12p(216-688)] from a GAL promoter causes FUS1 induction in a strain expressing wild-type STE12, suggesting that this region can cause the activation of endogenous Ste12p. Residues 262 to 594 are sufficient to cause STE12-dependent FUS1 induction when overexpressed, and this region of Ste12p was found to bind Dig1p but not Dig2p in yeast extracts. In contrast, recombinant glutathione S-transferase-Dig2p binds to the Ste12p DNA-binding domain (DBD). Expression of DIG2, but not DIG1, from a GAL promoter inhibits transcriptional activation by an Ste12p DBD-VP16 fusion. Furthermore, disruption of dig1, but not dig2, causes elevated transcriptional activation by a LexA-Ste12p(216-688) fusion. Ste12p has multiple regions within the C terminus (flanking residue 474) that can promote multimerization in vitro, and we demonstrate that these interactions can contribute to the activation of endogenous Ste12p by overproduced C-terminal fragments. These results demonstrate that Dig1p and Dig2p do not function by redundant mechanisms but rather inhibit pheromone-responsive transcription through interactions with separate regions of Ste12p. PMID- 10825186 TI - p53 regulation of G(2) checkpoint is retinoblastoma protein dependent. AB - In the present study, we investigated the role of p53 in G(2) checkpoint function by determining the mechanism by which p53 prevents premature exit from G(2) arrest after genotoxic stress. Using three cell model systems, each isogenic, we showed that either ectopic or endogenous p53 sustained a G(2) arrest activated by ionizing radiation or adriamycin. The mechanism was p21 and retinoblastoma protein (pRB) dependent and involved an initial inhibition of cyclin B1-Cdc2 activity and a secondary decrease in cyclin B1 and Cdc2 levels. Abrogation of p21 or pRB function in cells containing wild-type p53 blocked the down-regulation of cyclin B1 and Cdc2 expression and led to an accelerated exit from G(2) after genotoxic stress. Thus, similar to what occurs in p21 and p53 deficiency, pRB loss can uncouple S phase and mitosis after genotoxic stress in tumor cells. These results indicate that similar molecular mechanisms are required for p53 regulation of G(1) and G(2) checkpoints. PMID- 10825187 TI - Interference footprinting analysis of telomerase elongation complexes. AB - Telomerase is a reverse transcriptase that adds single-stranded telomeric repeats to the ends of linear eukaryotic chromosomes. It consists of an RNA molecule including a template sequence, a protein subunit containing reverse transcriptase motifs, and auxiliary proteins. We have carried out an interference footprinting analysis of the Tetrahymena telomerase elongation complexes. In this study, single-stranded oligonucleotide primers containing telomeric sequences were modified with base-specific chemical reagents and extended with the telomerase by a single (32)P-labeled dGMP or dTMP. Base modifications that interfered with the primer extension reactions were mapped by footprinting. Major functional interactions were detected between the telomerase and the six or seven 3' terminal residues of the primers. These interactions occurred not only with the RNA template region, but also with another region in the enzyme ribonucleoprotein complex designated the telomerase DNA interacting surface (TDIS). This was indicated by footprints generated with dimethyl sulfate (that did not affect Watson-Crick hydrogen bonding) and by footprinting assays performed with mutant primers. In primers aligned at a distance of 2 nucleotides along the RNA template region, the footprints of the six or seven 3'-terminal residues were shifted by 2 nucleotides. This shift indicated that during the elongation reaction, TDIS moved in concert with the 3' ends of the primers relative to the template region. Weak interactions occurred between the telomerase and residues located upstream of the seventh nucleotide. These interactions were stronger in primers that were impaired in the ability to align with the template. PMID- 10825188 TI - Differential mechanisms of LEF/TCF family-dependent transcriptional activation by beta-catenin and plakoglobin. AB - beta-Catenin and plakoglobin are highly homologous components of cell-cell adherens junctions linking cadherin receptors to the actin cytoskeleton. beta Catenin, in addition, activates transcription by forming a complex with LEF/TCF family transcription factors in the nucleus. Plakoglobin can also bind to LEF-1 and, when overexpressed in mammalian cells, enhances LEF-1-directed transcription. Plakoglobin overexpression, however, results in the elevation and nuclear translocation of endogenous beta-catenin. We show here, by DNA mobility shift analysis, that the formation of a plakoglobin-LEF/TCF-DNA complex in vitro is very inefficient compared to a complex containing beta-catenin-LEF-DNA. Moreover, in plakoglobin-transfected cells plakoglobin-LEF/TCF-DNA complexes were not formed; rather, the endogenous beta-catenin, whose level is elevated by plakoglobin transfection, formed a beta-catenin-LEF-DNA complex. Removal of the N and C-terminal domains of both beta-catenin and plakoglobin (leaving the armadillo repeat domain intact) induced plakoglobin-LEF-DNA complex formation and also enhanced beta-catenin-LEF-DNA complexing, both with in vitro-translated components and in transfected cells. Transfection with these truncated catenins increased endogenous beta-catenin levels, but the truncated catenins acted as dominant-negative inhibitors of beta-catenin-driven transcription by forming transcriptionally inactive complexes with LEF-1. When these catenin mutants were prevented from entering the nucleus, by their fusion to the connexin transmembrane domain, they indirectly activated transcription by increasing endogenous beta-catenin levels. These results suggest that overexpression of plakoglobin does not directly activate transcription and that formation of catenin-LEF-DNA complexes is negatively regulated by the catenin N- and C terminal domains. PMID- 10825189 TI - Mad1 function is regulated through elements within the carboxy terminus. AB - Myc and Mad are basic helix-loop-helix leucine zipper (bHLH-LZ) proteins that heterodimerize with Max to bind DNA and thereby influence the transcription of Myc-responsive genes. Myc-Max dimers transactivate whereas Mad-Max-mSin3 complexes repress Myc-mediated transcriptional activation. We have previously shown that the N-terminal mSin3 binding domain and the centrally located bHLH-LZ are required for Mad1 to function during a molecular switch from proliferation to differentiation. Here we demonstrate that the carboxy terminus (CT) of Mad1 contains previously unidentified motifs necessary for the regulation of Mad1 function. We show that removal of the last 18 amino acids of Mad1 (region V) abolishes the growth-inhibitory function of the protein and the ability to reverse a Myc-imposed differentiation block. Moreover, deletion of region V results in a protein that binds DNA weakly and no longer represses Myc-dependent transcriptional activation. In contrast, deletion of the preceding 24 amino acids (region IV) together with region V restores DNA binding and transcriptional repression, suggesting a functional interplay between these two regions. Furthermore, phosphorylation within region IV appears to mediate this interplay. These findings indicate that novel regulatory elements are present in the Mad1 CT. PMID- 10825190 TI - Regulation of cyclooxygenase 2 mRNA stability by the mitogen-activated protein kinase p38 signaling cascade. AB - A tetracycline-regulated reporter system was used to investigate the regulation of cyclooxygenase 2 (Cox-2) mRNA stability by the mitogen-activated protein kinase (MAPK) p38 signaling cascade. The stable beta-globin mRNA was rendered unstable by insertion of the 2, 500-nucleotide Cox-2 3' untranslated region (3' UTR). The chimeric transcript was stabilized by a constitutively active form of MAPK kinase 6, an activator of p38. This stabilization was blocked by SB203580, an inhibitor of p38, and by two different dominant negative forms of MAPK activated protein kinase 2 (MAPKAPK-2), a kinase lying downstream of p38. Constitutively active MAPKAPK-2 was also able to stabilize chimeric beta-globin Cox-2 transcripts. The MAPKAPK-2 substrate hsp27 may be involved in stabilization, as beta-globin-Cox-2 transcripts were partially stabilized by phosphomimetic mutant forms of hsp27. A short (123-nucleotide) fragment of the Cox-2 3' UTR was necessary and sufficient for the regulation of mRNA stability by the p38 cascade and interacted with a HeLa protein immunologically related to AU rich element/poly(U) binding factor 1. PMID- 10825191 TI - Disruption of the 11-cis-retinol dehydrogenase gene leads to accumulation of cis retinols and cis-retinyl esters. AB - To elucidate the possible role of 11-cis-retinol dehydrogenase in the visual cycle and/or 9-cis-retinoic acid biosynthesis, we generated mice carrying a targeted disruption of the 11-cis-retinol dehydrogenase gene. Homozygous 11-cis retinol dehydrogenase mutants developed normally, including their retinas. There was no appreciable loss of photoreceptors. Recently, mutations in the 11-cis retinol dehydrogenase gene in humans have been associated with fundus albipunctatus. In 11-cis-retinol dehydrogenase knockout mice, the appearance of the fundus was normal and punctata typical of this human hereditary ocular disease were not present. A second typical symptom associated with this disease is delayed dark adaptation. Homozygous 11-cis-retinol dehydrogenase mutants showed normal rod and cone responses. 11-cis-Retinol dehydrogenase knockout mice were capable of dark adaptation. At bleaching levels under which patients suffering from fundus albipunctatus could be detected unequivocally, 11-cis retinol dehydrogenase knockout animals displayed normal dark adaptation kinetics. However, at high bleaching levels, delayed dark adaptation in 11-cis-retinol dehydrogenase knockout mice was noticed. Reduced 11-cis-retinol oxidation capacity resulted in 11-cis-retinol/13-cis-retinol and 11-cis-retinyl/13-cis retinyl ester accumulation. Compared with wild-type mice, a large increase in the 11-cis-retinyl ester concentration was noticed in 11-cis-retinol dehydrogenase knockout mice. In the murine retinal pigment epithelium, there has to be an additional mechanism for the biosynthesis of 11-cis-retinal which partially compensates for the loss of the 11-cis-retinol dehydrogenase activity. 11-cis Retinyl ester formation is an important part of this adaptation process. Functional consequences of the loss of 11-cis-retinol dehydrogenase activity illustrate important differences in the compensation mechanisms between mice and humans. We furthermore demonstrate that upon 11-cis-retinol accumulation, the 13 cis-retinol concentration also increases. This retinoid is inapplicable to the visual processes, and we therefore speculate that it could be an important catabolic metabolite and its biosynthesis could be part of a process involved in regulating 11-cis-retinol concentrations within the retinal pigment epithelium of 11-cis-retinol dehydrogenase knockout mice. PMID- 10825192 TI - Mechanism of caffeine-induced checkpoint override in fission yeast. AB - Mitotic checkpoints restrain the onset of mitosis (M) when DNA is incompletely replicated or damaged. These checkpoints are conserved between the fission yeast Schizosaccharomyces pombe and mammals. In both types of organisms, the methylxanthine caffeine overrides the synthesis (S)-M checkpoint that couples mitosis to completion of DNA S phase. The molecular target of caffeine was sought in fission yeast. Caffeine prevented activation of Cds1 and phosphorylation of Chk1, two protein kinases that enforce the S-M checkpoint triggered by hydroxyurea. Caffeine did not inhibit these kinases in vitro but did inhibit Rad3, a kinase that regulates Cds1 and Chk1. In accordance with this finding, caffeine also overrode the G(2)-M DNA damage checkpoint that requires Rad3 function. Rad3 coprecipitated with Cds1 expressed at endogenous amounts, a finding that supports the hypothesis that Rad3 is involved in direct activation of Cds1. PMID- 10825193 TI - Yeast Ran-binding protein 1 (Yrb1) shuttles between the nucleus and cytoplasm and is exported from the nucleus via a CRM1 (XPO1)-dependent pathway. AB - The RanGTP-binding protein RanBP1, which is located in the cytoplasm, has been implicated in release of nuclear export complexes from the cytoplasmic side of the nuclear pore complex. Here we show that Yrb1 (the yeast homolog of RanBP1) shuttles between the nucleus and the cytoplasm. Nuclear import of Yrb1 is a facilitated process that requires a short basic sequence within the Ran-binding domain (RBD). By contrast, nuclear export of Yrb1 requires an intact RBD, which forms a ternary complex with the Xpo1 (Crm1) NES receptor in the presence of RanGTP. Nuclear export of Yrb1, however, is insensitive towards leptomycin B, suggesting a novel type of substrate recognition between Yrb1 and Xpo1. Taken together, these data suggest that ongoing nuclear import and export is an important feature of Yrb1 function in vivo. PMID- 10825194 TI - The c-Myc transactivation domain is a direct modulator of apoptotic versus proliferative signals. AB - We have assayed the oncogenic, proliferative, and apoptotic activities of the frequent mutations that occur in the c-myc gene in Burkitt's lymphomas. Some alleles have a modest (50 to 60%) increase in transforming activity; however, the most frequent Burkitt's lymphoma allele (T58I) had an unexpected substantial decrease in transforming activity (85%). All alleles restored the proliferation function of c-Myc in cells that grow slowly due to a c-myc knockout. There was discordance for some alleles between apoptotic and oncogenic activities, but only the T58A allele had elevated transforming activity with a concomitant reduced apoptotic potential. We discovered a novel missense mutation, MycS71F, that had a very low apoptotic activity compared to wild-type Myc, yet this mutation has never been found in lymphomas, suggesting that there is no strong selection for antiapoptotic c-Myc alleles. MycS71F also induced very low levels of cytochrome c release from mitochondria, suggesting a mechanism of action for this mutation. Phosphopeptide mapping provided a biochemical basis for the dramatically different biological activities of the transformation-defective T58I and transformation-enhanced T58A c-Myc alleles. Furthermore, the antiapoptotic survival factor insulin-like growth factor 1 was found to suppress phosphorylation of T58, suggesting that the c-Myc transactivation domain is a direct target of survival signals. PMID- 10825195 TI - Characterization of a CREB gain-of-function mutant with constitutive transcriptional activity in vivo. AB - The cyclic AMP (cAMP)-responsive factor CREB promotes cellular gene expression, following its phosphorylation at Ser133, via recruitment of the coactivator paralogs CREB-binding protein (CBP) and p300. CBP and p300, in turn, appear to mediate target gene induction via their association with RNA polymerase II complexes and via intrinsic histone acetyltransferase activities that mobilize promoter-bound nucleosomes. In addition to cAMP, a wide variety of stimuli, including hypoxia, UV irradiation, and growth factor addition, induce Ser133 phosphorylation with stoichiometry and kinetics comparable to those induced by cAMP. Yet a number of these signals are incapable of promoting target gene activation via CREB phosphorylation per se, suggesting the presence of additional regulatory events either at the level of CREB-CBP complex formation or in the subsequent recruitment of the transcriptional apparatus. Here we characterize a Tyr134Phe CREB mutant that behaves as a constitutive activator in vivo. Like protein kinase A (PKA)-stimulated wild-type CREB, the Tyr134Phe polypeptide was found to stimulate target gene expression via the Ser133-dependent recruitment of CBP and p300. Biochemical studies reveal that mutation of Tyr134 to Phe lowers the K(m) for PKA phosphorylation and thereby induces high levels of constitutive Ser133 phosphorylation in vivo. Consistent with its constitutive activity, Tyr134Phe CREB strongly promoted differentiation of PC12 cells in concert with suboptimal doses of nerve growth factor. Taken together, these results demonstrate that Ser133 phosphorylation is sufficient for cellular gene activation and that additional signal-dependent modifications of CBP or p300 are not required for recruitment of the transcriptional apparatus to the promoter. PMID- 10825196 TI - Novel mechanism of steroid action in skin through glucocorticoid receptor monomers. AB - Glucocorticoids (GCs), important regulators of epidermal growth, differentiation, and homeostasis, are used extensively in the treatment of skin diseases. Using keratin gene expression as a paradigm of epidermal physiology and pathology, we have developed a model system to study the molecular mechanism of GCs action in skin. Here we describe a novel mechanism of suppression of transcription by the glucocorticoid receptor (GR) that represents an example of customizing a device for transcriptional regulation to target a specific group of genes within the target tissue, in our case, epidermis. We have shown that GCs repress the expression of the basal-cell-specific keratins K5 and K14 and disease-associated keratins K6, K16, and K17 but not the differentiation-specific keratins K3 and K10 or the simple epithelium-specific keratins K8, K18, and K19. We have identified the negative recognition elements (nGREs) in all five regulated keratin gene promoters. Detailed footprinting revealed that the function of nGREs is to instruct the GR to bind as four monomers. Furthermore, using cotransfection and antisense technology we have found that, unlike SRC-1 and GRIP-1, which are not involved in the GR complex that suppresses keratin genes, histone acetyltransferase and CBP are. In addition, we have found that GR, independently from GREs, blocks the induction of keratin gene expression by AP1. We conclude that GR suppresses keratin gene expression through two independent mechanisms: directly, through interactions of keratin nGREs with four GR monomers, as well as indirectly, by blocking the AP1 induction of keratin gene expression. PMID- 10825197 TI - Aca1 and Aca2, ATF/CREB activators in Saccharomyces cerevisiae, are important for carbon source utilization but not the response to stress. AB - In Saccharomyces cerevisiae, the family of ATF/CREB transcriptional regulators consists of a repressor, Acr1 (Sko1), and two activators, Aca1 and Aca2. The AP-1 factor Gen4 does not activate transcription through ATF/CREB sites in vivo even though it binds these sites in vitro. Unlike ATF/CREB activators in other species, Aca1- and Aca2-dependent transcription is not affected by protein kinase A or by stress, and Aca1 and Aca2 are not required for Hog1-dependent salt induction of transcription through an optimal ATF/CREB site. Aca2 is important for a variety of biological functions including growth on nonoptimal carbon sources, and Aca2-dependent activation is modestly regulated by carbon source. Strains lacking Aca1 are phenotypically normal, but overexpression of Aca1 suppresses some defects associated with the loss of Aca2, indicating a functional overlap between Aca1 and Aca2. Acr1 represses transcription both by recruiting the Cyc8-Tup1 corepressor and by directly competing with Aca1 and Aca2 for target sites. Acr1 does not fully account for osmotic regulation through ATF/CREB sites, and a novel Hog1-dependent activator(s) that is not a bZIP protein is required for ATF/CREB site activation in response to high salt. In addition, Acr1 does not affect a number of phenotypes that arise from loss of Aca2. Thus, members of the S. cerevisiae ATF/CREB family have overlapping, but distinct, biological functions and target genes. PMID- 10825198 TI - Artificial recruitment of TFIID, but not RNA polymerase II holoenzyme, activates transcription in mammalian cells. AB - In yeast cells, transcriptional activation occurs when the RNA polymerase II (Pol II) machinery is artificially recruited to a promoter by fusing individual components of this machinery to a DNA-binding domain. Here, we show that artificial recruitment of components of the TFIID complex can activate transcription in mammalian cells. Surprisingly, artificial recruitment of TATA binding protein (TBP) activates transiently transfected and chromosomally integrated promoters with equal efficiency, whereas artificial recruitment of TBP associated factors activates only chromosomal reporters. In contrast, artificial recruitment of various components of the mammalian Pol II holoenzyme does not confer transcriptional activation, nor does it result in synergistic activation in combination with natural activation domains. In the one case examined in more detail, the Srb7 fusion failed to activate despite being associated with the Pol II holoenzyme and being directly recruited to the promoter. Interestingly, some acidic activation domains are less effective when the promoter is chromosomally integrated rather than transiently transfected, whereas the Sp1 glutamine-rich activation domain is more effective on integrated reporters. Thus, yeast and mammalian cells differ with respect to transcriptional activation by artificial recruitment of the Pol II holoenzyme. PMID- 10825200 TI - Hierarchy of protein tyrosine kinases in interleukin-2 (IL-2) signaling: activation of syk depends on Jak3; however, neither Syk nor Lck is required for IL-2-mediated STAT activation. AB - Interleukin-2 (IL-2) activates several different families of tyrosine kinases, but precisely how these kinases interact is not completely understood. We therefore investigated the functional relationships among Jak3, Lck, and Syk in IL-2 signaling. We first observed that in the absence of Jak3, both Lck and Syk had the capacity to phosphorylate Stat3 and Stat5a. However, neither supported IL 2-induced STAT activation, nor did dominant negative alleles of these kinases inhibit. Moreover, pharmacological abrogation of Lck activity did not inhibit IL 2-mediated phosphorylation of Jak3 and Stat5a. Importantly, ligand-dependent Syk activation was dependent on the presence of catalytically active Jak3, whereas Lck activation was not. Interestingly, Syk functioned as a direct substrate of Jak1 but not Jak3. Additionally, Jak3 phosphorylated Jak1, whereas the reverse was not the case. Taken together, our data support a model in which Lck functions in parallel with Jak3, while Syk functions as a downstream element of Jaks in IL 2 signaling. Jak3 may regulate Syk catalytic activity indirectly via Jak1. However, IL-2-mediated Jak3/Stat activation is not dependent on Lck or Syk. While the essential roles of Jak1 and Jak3 in signaling by gammac-utilizing cytokines are clear, it will be important to dissect the exact contributions of Lck and Syk in mediating the effects of IL-2 and related cytokines. PMID- 10825199 TI - Mechanism for specificity by HMG-1 in enhanceosome assembly. AB - Assembly of enhanceosomes requires architectural proteins to facilitate the DNA conformational changes accompanying cooperative binding of activators to a regulatory sequence. The architectural protein HMG-1 has been proposed to bind DNA in a sequence-independent manner, yet, paradoxically, it facilitates specific DNA binding reactions in vitro. To investigate the mechanism of specificity we explored the effect of HMG-1 on binding of the Epstein-Barr virus activator ZEBRA to a natural responsive promoter in vitro. DNase I footprinting, mutagenesis, and electrophoretic mobility shift assay reveal that HMG-1 binds cooperatively with ZEBRA to a specific DNA sequence between two adjacent ZEBRA recognition sites. This binding requires a strict alignment between two adjacent ZEBRA sites and both HMG boxes of HMG-1. Our study provides the first demonstration of sequence dependent binding by a nonspecific HMG-box protein. We hypothesize how a ubiquitous, nonspecific architectural protein can function in a specific context through the use of rudimentary sequence recognition coupled with cooperativity. The observation that an abundant architectural protein can bind DNA cooperatively and specifically has implications towards understanding HMG-1's role in mediating DNA transactions in a variety of enzymological systems. PMID- 10825201 TI - The CaaX proteases, Afc1p and Rce1p, have overlapping but distinct substrate specificities. AB - Many proteins that contain a carboxyl-terminal CaaX sequence motif, including Ras and yeast a-factor, undergo a series of sequential posttranslational processing steps. Following the initial prenylation of the cysteine, the three C-terminal amino acids are proteolytically removed, and the newly formed prenylcysteine is carboxymethylated. The specific amino acids that comprise the CaaX sequence influence whether the protein can be prenylated and proteolyzed. In this study, we evaluated processing of a-factor variants with all possible single amino acid substitutions at either the a(1), the a(2), or the X position of the a-factor Ca(1)a(2)X sequence, CVIA. The substrate specificity of the two known yeast CaaX proteases, Afc1p and Rce1p, was investigated in vivo. Both Afc1p and Rce1p were able to proteolyze a-factor with A, V, L, I, C, or M at the a(1) position, V, L, I, C, or M at the a(2) position, or any amino acid at the X position that was acceptable for prenylation of the cysteine. Eight additional a-factor variants with a(1) substitutions were proteolyzed by Rce1p but not by Afc1p. In contrast, Afc1p was able to proteolyze additional a-factor variants that Rce1p may not be able to proteolyze. In vitro assays indicated that farnesylation was compromised or undetectable for 11 a-factor variants that produced no detectable halo in the wild-type AFC1 RCE1 strain. The isolation of mutations in RCE1 that improved proteolysis of a-factor-CAMQ, indicated that amino acid substitutions E139K, F189L, and Q201R in Rce1p affected its substrate specificity. PMID- 10825203 TI - Cloning and functional characterization of the early-lymphocyte-specific Pb99 gene. AB - The Pb99 gene is specifically expressed in pre-B cells and thymocytes and not in mature B and T cells or nonlymphoid tissues, implying that it may function in early lymphoid development. We have previously described the cloning of an incomplete cDNA for Pb99. Here we report the isolation of full-length cDNAs and genomic clones for the murine Pb99 gene and the mapping of its location to mouse chromosome 8. Sequence analyses of different Pb99 cDNA clones suggest that there may be at least three forms of the Pb99 protein generated by differential processing of the Pb99 transcript. The cDNA with the longest open reading frame encodes a putative protein that has seven hydrophobic domains similar to those of seven membrane-spanning proteins, such as the classical G protein-coupled receptors. To directly address the role of the Pb99 protein in lymphoid development, Pb99-deficient mice were generated by gene targeting, and lymphocyte development in these mice was analyzed. PMID- 10825202 TI - DNA repair protein Rad55 is a terminal substrate of the DNA damage checkpoints. AB - Checkpoints, which are integral to the cellular response to DNA damage, coordinate transient cell cycle arrest and the induced expression of DNA repair genes after genotoxic stress. DNA repair ensures cellular survival and genomic stability, utilizing a multipathway network. Here we report evidence that the two systems, DNA damage checkpoint control and DNA repair, are directly connected by demonstrating that the Rad55 double-strand break repair protein of the recombinational repair pathway is a terminal substrate of DNA damage and replication block checkpoints. Rad55p was specifically phosphorylated in response to DNA damage induced by the alkylating agent methyl methanesulfonate, dependent on an active DNA damage checkpoint. Rad55p modification was also observed after gamma ray and UV radiation. The rapid time course of phosphorylation and the recombination defects identified in checkpoint-deficient cells are consistent with a role of the DNA damage checkpoint in activating recombinational repair. Rad55p phosphorylation possibly affects the balance between different competing DNA repair pathways. PMID- 10825204 TI - Sli2 (Ypk1), a homologue of mammalian protein kinase SGK, is a downstream kinase in the sphingolipid-mediated signaling pathway of yeast. AB - ISP-1 is a new type of immunosuppressant, the structure of which is homologous to that of sphingosine. In a previous study, ISP-1 was found to inhibit mammalian serine palmitoyltransferase, the primary enzyme involved in sphingolipid biosynthesis, and to reduce the intracellular pool of sphingolipids. ISP-1 induces the apoptosis of cytotoxic T cells, which is triggered by decreases in the intracellular levels of sphingolipids. In this study, the inhibition of yeast (Saccharomyces cerevisiae) proliferation by ISP-1 was observed. This ISP-1 induced growth inhibition was also triggered by decreases in the intracellular levels of sphingolipids. In addition, DNA duplication without cytokinesis was detected in ISP-1-treated yeast cells on flow cytometry analysis. We have cloned multicopy suppressor genes of yeast which overcome the lethal sphingolipid depletion induced by ISP-1. One of these genes, SLI2, is synonymous with YPK1, which encodes a serine/threonine kinase. Kinase-dead mutants of YPK1 did not show any resistance to ISP-1, leading us to predict that the kinase activity of the Ypk1 protein should be essential for this resistance to ISP-1. Ypk1 protein overexpression had no effect on sphingolipid biosynthesis by the yeast. Furthermore, both the phosphorylation and intracellular localization of the Ypk1 protein were regulated by the intracellular sphingolipid levels. These data suggest that the Ypk1 protein is a downstream kinase in the sphingolipid-mediated signaling pathway of yeast. The Ypk1 protein was reported to be a functional homologue of the mammalian protein kinase SGK, which is a downstream kinase of 3 phosphoinositide-dependent kinase 1 (PDK1). PDK1 phosphotidylinositol (PI) is regulated by PI-3,4,5-triphosphate and PI-3,4-bisphosphate through the pleckstrin homology (PH) domain. Overexpression of mammalian SGK also overcomes the sphingolipid depletion in yeast. Taking both the inability to produce PI-3,4, 5 triphosphate and PI-3,4-bisphosphate and the lack of a PH domain in the yeast homologue of PDK1, the Pkh1 protein, into account, these findings further suggest that yeast may use sphingolipids instead of inositol phospholipids as lipid mediators. PMID- 10825205 TI - Replication delay along FRA7H, a common fragile site on human chromosome 7, leads to chromosomal instability. AB - Common fragile sites are specific chromosomal loci that show gaps, breaks, or rearrangements in metaphase chromosomes under conditions that interfere with DNA replication. The mechanism underlying the chromosomal instability at fragile sites was hypothesized to associate with late replication time. Here, we aimed to investigate the replication pattern of the common fragile site FRA7H, encompassing 160 kb on the long arm of human chromosome 7. Using in situ hybridization on interphase nuclei, we revealed that the replication of this region is initiated relatively early, before 30% of S phase is completed. However, a high fraction ( approximately 35%) of S-phase nuclei showed allelic asynchrony, indicating that the replication of FRA7H is accomplished at different times in S phase. This allelic asynchrony is not the result of a specific replication time of each FRA7H allele. Analysis of the replication pattern of adjacent clones along FRA7H by using cell population and two-color fluorescent in situ hybridization analyses showed significant differences in the replication of adjacent clones, under normal growth condition and upon aphidicolin treatment. This pattern significantly differed from that of two nonfragile regions which showed a coordinated replication under both conditions. These results indicate that aphidicolin is enhancing an already existing difference in the replication time along the FRA7H region. Based on our replication analysis of FRA7H and on previous analysis of the common fragile site FRA3B, we suggest that delayed replication is underlying the fragility at aphidicolin-induced common fragile sites. PMID- 10825206 TI - A zinc finger transcription factor, alphaA-crystallin binding protein 1, is a negative regulator of the chondrocyte-specific enhancer of the alpha1(II) collagen gene. AB - Transcription of the type II collagen gene (Col2a1) is regulated by multiple cis acting sites. The enhancer element, which is located in the first intron, is necessary for high-level and cartilage-specific expression of Col2a1. A mouse limb bud cDNA expression library was screened by the Saccharomyces cerevisiae one hybrid screening method to identify protein factors bound to the enhancer. A zinc finger protein, alphaA-crystallin binding protein 1 (CRYBP1), which had been reported to bind to the mouse alphaA-crystallin gene promoter, was isolated. We herein demonstrate that CRYBP1 is involved in the negative regulation of Col2a1 enhancer activity. CRYBP1 mRNA expression was downregulated during chondrocyte differentiation in vitro. In situ hybridization analysis of developing mouse cartilage showed that CRYBP1 mRNA was also downregulated during mesenchymal condensation and that CRYBP1 mRNA was highly expressed by hypertrophic chondrocytes, but at very low levels by resting and proliferating chondrocytes. Expression of recombinant CRYBP1 in a transfected rat chondrosarcoma cell line inhibited Col2a1 enhancer activity. Electrophoretic mobility shift assays showed that CRYBP1 bound a specific sequence within the Col2a1 enhancer and inhibited the binding of Sox9, an activator for Col2a1, to the enhancer. Cotransfection of CRYBP1 with Sox9 into BALB/c 3T3 cells inhibited activation of the Col2a1 enhancer by Sox9. These results suggest a novel mechanism that negatively regulates cartilage-specific expression of Col2a1. PMID- 10825207 TI - Hepatocyte-specific mutation establishes retinoid X receptor alpha as a heterodimeric integrator of multiple physiological processes in the liver. AB - A large number of physiological processes in the adult liver are regulated by nuclear receptors that require heterodimerization with retinoid X receptors (RXRs). In this study, we have used cre-mediated recombination to disrupt the mouse RXRalpha gene specifically in hepatocytes. Although such mice are viable, molecular and biochemical parameters indicate that every one of the examined metabolic pathways in the liver (mediated by RXR heterodimerization with PPARalpha, CARbeta, PXR, LXR, and FXR) is compromised in the absence of RXRalpha. These data demonstrate the presence of a complex circuitry in which RXRalpha is integrated into a number of diverse physiological pathways as a common regulatory component of cholesterol, fatty acid, bile acid, steroid, and xenobiotic metabolism and homeostasis. PMID- 10825208 TI - Transcription factor hepatocyte nuclear factor 6 regulates pancreatic endocrine cell differentiation and controls expression of the proendocrine gene ngn3. AB - Hepatocyte nuclear factor 6 (HNF-6) is the prototype of a new class of cut homeodomain transcription factors. During mouse development, HNF-6 is expressed in the epithelial cells that are precursors of the exocrine and endocrine pancreatic cells. We have investigated the role of HNF-6 in pancreas differentiation by inactivating its gene in the mouse. In hnf6(-/-) embryos, the exocrine pancreas appeared to be normal but endocrine cell differentiation was impaired. The expression of neurogenin 3 (Ngn-3), a transcription factor that is essential for determination of endocrine cell precursors, was almost abolished. Consistent with this, we demonstrated that HNF-6 binds to and stimulates the ngn3 gene promoter. At birth, only a few endocrine cells were found and the islets of Langerhans were missing. Later, the number of endocrine cells increased and islets appeared. However, the architecture of the islets was perturbed, and their beta cells were deficient in glucose transporter 2 expression. Adult hnf6(-/-) mice were diabetic. Taken together, our data demonstrate that HNF-6 controls pancreatic endocrine differentiation at the precursor stage and identify HNF-6 as the first positive regulator of the proendocrine gene ngn3 in the pancreas. They also suggest that HNF-6 is a candidate gene for diabetes mellitus in humans. PMID- 10825209 TI - Mutations in the bare lymphocyte syndrome define critical steps in the assembly of the regulatory factor X complex. AB - The regulatory factor X (RFX) complex, which contains RFXANK(B), RFXAP, and RFX5, binds to X and S boxes in major histocompatibility complex class II (MHC II) promoters. In the bare lymphocyte syndrome (BLS), which is a human severe combined immunodeficiency, MHC II promoters are neither occupied nor transcribed. Thus, the absence of any one subunit prevents the formation of the RFX complex. Nevertheless, except for a weak binding between RFX5 and RFXAP, no other interactions between RFX proteins have been described. In this study, we demonstrate that RFXANK(B) binds to RFXAP to form a scaffold for the assembly of the RFX complex, which then binds to DNA. Moreover, mutant RFXANK(B) and RFXAP proteins from complementation groups B and D of BLS, respectively, cannot support this interaction. Our data elucidate an intriguing medical situation, where a genetic disease targets two different surfaces that are required for the nucleation of a multisubunit DNA-protein complex. PMID- 10825212 TI - Prevention of monocyte adhesion and inflammatory cytokine production during blood platelet storage: an in vitro model with implications for transfusion practice. AB - A novel platelet additive solution [ThromboSoltrade mark (TS)] was designed to allow extended refrigerated platelet storage. It has been shown to preserve platelet function and prevent cytokine accumulation in platelet concentrates stored for up to 9 days. It consists of amiloride, adenosine, sodium nitroprusside, dipyridamole, quinacrine, and ticlopidine. We hypothesized that the cytokine inhibition may be due to prevention of monocyte (MC) adhesion and activation on the surfaces of platelet storage bag plastic polymers. In an in vitro model, we incubated purified peripheral blood MCs on discs of polyolefin and polyvinylchloride from platelet storage bags, and on polystyrene, in the presence of TS for up to 7 days. We found that after incubation with TS, adherent MC numbers were decreased by >80-95% compared with controls on all surfaces examined. Levels of cytokines [interleukin (IL)-1beta, IL-1RA, IL-6, IL-8, and tumor necrosis factor-alpha] were low in wells with TS but rose progressively in the controls during incubation. Amiloride alone had similar effects on adhesion and cytokine release as the complete TS preparation. Removing amiloride from TS abrogated these effects. These findings suggest an important role for TS and amiloride in monocyte function, and have implications for the development of agents designed for prolonged platelet storage. PMID- 10825211 TI - Disruption of the murine calpain small subunit gene, Capn4: calpain is essential for embryonic development but not for cell growth and division. AB - Calpains are a family of Ca(2+)-dependent intracellular cysteine proteases, including the ubiquitously expressed micro- and m-calpains. Both mu- and m calpains are heterodimers, consisting of a distinct large 80-kDa catalytic subunit, encoded by the genes Capn1 and Capn2, and a common small 28-kDa regulatory subunit (Capn4). The physiological roles and possible functional distinctions of mu- and m-calpains remain unclear, but suggested functions include participation in cell division and migration, integrin-mediated signal transduction, apoptosis, and regulation of cellular control proteins such as cyclin D1 and p53. Homozygous disruption of murine Capn4 eliminated both mu- and m-calpain activities, but this did not affect survival and proliferation of cultured embryonic stem cells or embryonic fibroblasts, or the early stages of organogenesis. However, mutant embryos died at midgestation and displayed defects in the cardiovascular system, hemorrhaging, and accumulation of erythroid progenitors. PMID- 10825210 TI - The oncoprotein kinase chaperone CDC37 functions as an oncogene in mice and collaborates with both c-myc and cyclin D1 in transformation of multiple tissues. AB - CDC37 encodes a 50-kDa protein that targets intrinsically unstable oncoprotein kinases including Cdk4, Raf-1, and v-src to the molecular chaperone Hsp90, an interaction that is thought to be important for the establishment of signaling pathways. CDC37 is required for proliferation in budding yeast and is coexpressed with cyclin D1 in proliferative zones during mouse development, a finding consistent with a positive role in cell proliferation. CDC37 expression may not only be required to support proliferation in cells that are developmentally programmed to proliferate but may also be required in cells that are inappropriately induced to initiate proliferation by oncogenes. Here we report that mouse mammary tumor virus (MMTV)-CDC37 transgenic mice develop mammary gland tumors at a rate comparable to that observed previously in MMTV-cyclin D1 mice. Moreover, CDC37 was found to collaborate with MMTV-c-myc in the transformation of multiple tissues, including mammary and salivary glands in females and testis in males, and also collaborates with cyclin D1 to transform the female mammary gland. These data indicate that CDC37 can function as an oncogene in mice and suggests that the establishment of protein kinase pathways mediated by Cdc37 Hsp90 can be a rate-limiting event in epithelial cell transformation. PMID- 10825213 TI - NIST workshop on needs for reference biomaterials. PMID- 10825214 TI - Preparation of betaTCP/HAP biphasic ceramics with natural bone structure by heating bovine cancellous bone with the addition of (NH(4))(2)HPO(4). AB - In this study, the calcined bovine bone (CBB)-removing the organic substance by a burning process-with addition of different quantities of ammonium phosphate [(NH(4))(2)HPO(4)] (AP) was heated to a high temperature to transform its crystalline phase constitution from hydroxyapatite (HAP) into a tricalcium phosphate (TCP)/HAP biphasic structure. Results revealed that the CBB without AP appeared to be mainly composed of an HAP type pattern when heated to 1300 degrees C. After adding doped AP to CBB, the HPO(4)(2-) ions of AP condensed into P(2)O(7)(4-) ions at temperatures of 400-600 degrees C. P(2)O(7)(4-) ions reacted with the OH(-) ions of HAP to form betaTCP at temperatures up to 600 degrees C. The conversion reaction of HAP to betaTCP finished at around 900 degrees C. With increasing AP in the CBB, HAP gradually converted into different phase compositions of TCP/HAP or TCP at high temperature. Mechanical testing results showed that there was no significant difference in sintered CBB with different quantities of AP. By heating calcined bovine cancellous bone with different quantities of AP, we obtained different crystalline phase compositions of bioceramics with a natural porous structure. PMID- 10825215 TI - Photoencapsulation of chondrocytes in poly(ethylene oxide)-based semi interpenetrating networks. AB - A photopolymerizing hydrogel system provides an efficient method to encapsulate cells. The present work describes the in vitro analysis of bovine and ovine chondrocytes encapsulated in a poly(ethylene oxide)-dimethacrylate and poly(ethylene glycol) semi-interpenetrating network using a photopolymerization process. One day after encapsulation, (3-[4,5-dimethylthiazol-2-y1]-2, 5-diphenyl 2H-tetrazolium bromide) (MTT) and light microscopy showed chondrocyte survival and a dispersed cell population composed of ovoid and elongated cells. Biochemical analysis demonstrated proteoglycan and collagen contents that increased over 2 weeks of static incubation. Cell content of the gels initially decreased and stabilized. Biomechanical analysis demonstrated the presence of a functional extracellular matrix with equilibrium moduli, dynamic stiffness, and streaming potentials that increased with time. These findings suggest the feasibility of photoencapsulation for tissue engineering and drug delivery purposes. PMID- 10825216 TI - Effects of fatigue loading and PMMA precoating on the adhesion and subcritical debonding of prosthetic-PMMA interfaces. AB - Debonding of clinically relevant CoCrMo-polymethylmethacrylate (PMMA) interfaces is shown to occur subcritically under fatigue loading, implying that debonding may occur at loads much lower than those required for catastrophic failure. Interface fracture mechanics samples containing precoated and uncoated grit blasted CoCrMo substrates and a PMMA layer were constructed and quantitatively evaluated in terms of their critical interface adhesion and subcritical debond behavior. The precoat surfaces had markedly enhanced adhesion and fatigue resistance in both air and simulated physiological environmental conditions compared to the uncoated samples. Constraint of the PMMA layer does not significantly affect the debond process for thickness between 2- and 5-mm. In addition, wear particles were collected and shown to be consistent with particle sizes reported in vivo and are on the scale of the metal surface roughness. Life prediction methods using the subcritical debond-growth data are discussed. PMID- 10825217 TI - Targeted chemoembolization of tumors with poly(2-hydroxyethyl methacrylate) particles. AB - Thirty-four patients suffering from various kinds of tumors, including metastasis, were treated by selective embolization with both spherical and cylindrical poly(2-hydroxyethyl methacrylate) [poly(HEMA)] particles and topical chemotherapy. Treatment of a patient with carcinoid metastases in the liver is discussed. Immediately after embolization, 5-fluorouracil, and later, doxorubicin and Lipiodol, were selectively infused into the tumorous tissue for approximately 1 week. Patient received four cycles of this infusion. Chemoembolization proceeded against the background of anticoagulant therapy using small doses of heparin or its low-molecular-weight analogue, dalteparin. This was followed by transcutaneous transhepatic portography and embolization. Finally, the tumor feeding artery and portal vein were sealed by a hydrogel. After 1.5 months, the affected liver lobe was resected. Although 4 years from the beginning of treatment, the patient is still alive. Embolization with poly(HEMA) hydrogel particles in conjunction with an anticancer drug infusion via catheter is recommended as an efficient method of tumor treatment. The therapeutic effect has been shown to be a function of ischemia and slow local infusion of drug into the tumor, and systemic drug levels can be kept low. PMID- 10825218 TI - Effect of the continuous solution exchange on the in vitro reactivity of a CaO SiO(2) sol-gel glass. AB - The in vitro reactivity of a sol-gel glass with a composition in mol % of 80% SiO(2)-20% CaO (80S20C) was studied by soaking in a simulated body fluid (SBF). To model the continuous flow of body fluids, in this article a protocol for in vitro tests with continuous exchange of the assay solution (continuous) is proposed. The in vitro behavior of 80S20C in continuous is compared with that without SBF exchange (static). In static, remarkable variations in ionic concentration and pH of solution were detected after a few minutes of soaking. However, exchange of solution with 1 mL/min flow allowed us to maintain SBF ionic concentration and pH almost constant and close to plasma. Glass surface before and after soaking was studied by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDS), transmission electron microscopy (TEM), and electron diffraction (ED). After soaking, a calcium phosphate layer formed in both cases on the glass surface. However, some differences were observed as a function of the in vitro protocol used. In static, faster formation of the phosphate layer was detected in the first 6 h by FTIR, but for higher soaking times the situation was equivalent in both cases. After 7-day assay in continuous, XRD, SEM, TEM, and ED studies showed larger crystalline aggregates and apatite crystals on the newly formed layer. The use of a continuous protocol allowed us to perform complete in vitro studies with an ionic concentration and pH in solution almost identical to physiological fluids. PMID- 10825219 TI - Effect of aging time of sol on structure and in vitro calcium phosphate formation of sol-gel-derived titania films. AB - Titanium and its alloys have been used successfully in the manufacture of orthopedic and dental implants to replace damaged bone tissue. In this study, different sol-gel-derived TiO(2) coatings were produced on titanium substrates using different aging times (5, 10, 24, or 48 h) of the sol before dipping the coatings and varying numbers (one, three, or five) of coating layers. The influence of the aging time of the sol on the structure of the titania coatings with respect to in vitro bioactivity was investigated. The in vitro bioactivity tests were done in a simulated body fluid (SBF). The sol properties were monitored using a capillary viscometer and dynamic light scattering to determine the viscosity and particle size, respectively. The topography of the films was characterized using atomic force microscopy. The various sol aging times and numbers of layers produced differences in the topography of the titania films. For the coatings with one and three layers, the aging of the sols had an influence on the height of the peaks (lower with longer aging times) although the peak distance was about the same. The number of coating layers had a stronger influence. The distribution of the peak distances became narrower with an increasing number of coating layers. The coating with three layers (top coating prepared after 24 h of sol aging) and the coatings with five layers had a similar distribution of peak distances (15-50 nm), which was favorable for calcium phosphate formation. On these substrates, calcium phosphate formation started within 3-6 days of immersion in SBF. The aging time of the titania sol and the number of coating layers were found to have a strong influence on the surface topography in the nanometer scale of the titania films. The results indicate that the topography of the outermost surface is of importance for in vitro bioactivity. PMID- 10825220 TI - Covalent surface chemical modification of electrodes for cardiac pacing applications. AB - We report the covalent surface modification of active-fixation pacemaker electrodes with butanethiol or dodecanethiol self-assembled monolayers (SAMs) using a platinum or gold metal-thiolate bond (i.e., changing the chemical identity of the exposed metal electrode from metal to organic) in such fashion that (a) the surface is organic in functionality with lipophilic physicochemical characteristics, (b) a possible degradation product is gold (I)-alkanethiolate with putative anti-inflammatory actions, and (c) current density/electric field strength is increased. Superior acute and chronic pacing performance with dodecanethiol-modified, gold-coated, platinum-iridium alloy pacemaker electrodes was observed with inferential evidence of reduced inflammation and scar. This approach may have applicability in other areas of bioelectrodes with practical applications in clinical cardiology, surgery, neuroscience, and subcutaneous sensors. PMID- 10825221 TI - Temperature-responsive culture dishes allow nonenzymatic harvest of differentiated Madin-Darby canine kidney (MDCK) cell sheets. AB - We have developed a temperature-responsive culture dish grafted with a poly(N isopropylacrylamide) (PIPAAm). Various types of cells adhere, spread, and proliferate on the grafted dishes in the presence of serum at 37 degrees C. By reducing only temperature, these cells can be harvested noninvasively from the dishes according to rapid hydration of the grafted polymer. Because the harvest does not need enzymatic digestion, differentiated cell phenotypes are retained. In the present study, a renal epithelial cell line, Madin-Darby canine kidney (MDCK) cell, was cultured on the dishes, and cell behavior was examined. MDCK cells showed differentiated phenotypes such as dome formation during long-term culture, similar to on ungrafted dishes. After 1-week culture at 37 degrees C, trypsin digestion disrupted cell-cell junctions but failed to liberate cells from both ungrafted and grafted dishes. However, short-term incubation at 20 degrees C released confluent MDCK cells as a single contiguous cell sheet only from the polymer-grafted dishes because of selective disruption of the cell-surface binding. Immunocytochemistry with anti-beta-catenin antibody revealed that functional cell-cell junctions were organized even in the recovered cell sheets. Intriguingly, incubation time at 20 degrees C required for cell sheet detachment gradually shortened during long-term culture before reducing temperature. The acceleration of cell detachment was correlated to the decrease of a single cell area by means of cell contractile force. These findings suggest that cell sheet detachment from PIPAAm-grafted dishes should be accomplished by both PIPAAm hydration and cellular metabolic activity such as cell contraction. PMID- 10825222 TI - Pooled human immunoglobulins reduce adhesion of Pseudomonas aeruginosa in a parallel plate flow chamber. AB - The influence of pooled polyclonal immunoglobulin (IgG) interactions with both bacteria and model substrates in altering Pseudomonas aeruginosa surface adhesion is reported. Opsonization of this pathogen by polyclonal human IgG and preadsorption of IgG to glass surfaces both effectively reduce initial deposition rates and surface growth of P. aeruginosa IFO3455 from dilute nutrient broth in a parallel plate flow chamber. Polyclonal IgG depleted of P. aeruginosa-specific antibodies reduces the initial deposition rate or surface growth to levels intermediate between exposed and nonexposed IgG conditions. Bacterial surface properties are changed in the presence of opsonizing IgG. Plateau contact angle analysis via sessile drop technique shows a drop in P. aeruginosa surface hydrophobicity after IgG exposure consistent with a more hydrophilic IgG surface coat. Zeta potential values for opsonized versus nonopsonized bacteria exhibit little change. X-ray photoelectron spectroscopy measurements provide surface compositional evidence for IgG attachment to bacterial surfaces. Surface elemental ratios attributed to IgG protein signals versus those attributed primarily to bacterial polysaccharide surface or lipid membrane change with IgG opsonization. Direct evidence for antibody-modified P. aeruginosa surface properties correlates both with reduction of bacterial adhesion to glass surfaces under flow in nutrient medium reported and previous reports of IgG efficacy against P. aeruginosa motility in vitro and infection in vivo. PMID- 10825223 TI - In vitro characterization of mesenchymal stem cell-seeded collagen scaffolds for tendon repair: effects of initial seeding density on contraction kinetics. AB - Mesenchymal stem cells (MSCs) were isolated from bone marrow, culture-expanded, and then seeded at 1, 4, and 8 million cells/mL onto collagen gel constructs designed to augment tendon repair in vivo. To investigate the effects of seeding density on the contraction kinetics and cellular morphology, the contraction of the cell/collagen constructs was monitored over time up to 72 h in culture conditions. Constructs seeded at 4 and 8 million cells/mL showed no significant differences in their gross appearance and dimensions throughout the contraction process. By contrast, constructs seeded at 1 million cells/mL initially contracted more slowly and their diameters at 72 h were 62 to 73% larger than those seeded at higher densities. During contraction, MSCs reoriented and elongated significantly with time. Implants prepared at higher seeding densities showed more well aligned and elongated cell nuclei after 72 h of contraction. Changes in nuclear morphology of the MSCs in response to physical constraints provided by the contracted collagen fibrils may trigger differentiation pathways toward the fibroblastic lineage and influence the cell synthetic activity. Controlling the contraction and organization of the cells and matrix will be critical for successfully creating tissue engineered grafts. PMID- 10825224 TI - Immunohistochemical study of dental pulp applied with 4-META/MMA-TBB adhesive resin after pulpotomy. AB - The purpose of this study was to investigate nerve regeneration and proliferative activity in amputated pulp tissue after the application of 4-META/MMA-TBB adhesive resin (4-META resin). Calcium hydroxide was used as a control material. At 3 days, fibroblast-like cells were positive for proliferating cell nuclear antigen (PCNA) in both 4-META resin- and calcium hydroxide-treated groups and were located mainly within 0.5 mm from the cut surface. Only a few fragmented neurofilament protein (NFP)-positive nerve fibers were observed in this area. At 7 and 14 days, the number of PCNA-positive cells had gradually decreased and regenerated NFP-positive nerve fibers were observed close to the cut surface of the pulp in both groups. At 21 days in the experimental group, several PCNA positive cells were still found in the area 0.5 mm from the cut surface, and NFP positive nerve fibers were detected about 0.15-;0.2 mm from the cut surface. In contrast, a dentin bridge was produced under the necrotic layer at 21 days in the control group. PCNA-positive cells were not found underneath the dentin bridge, but NFP-positive nerve fibers had regenerated close to it. These results suggest that although cell differentiation and nerve regeneration are delayed, wound healing occurred even after the application of 4-META resin to exposed pulp surface the same as calcium hydroxide application. PMID- 10825225 TI - Albumin and heparin multilayer coatings for blood-contacting medical devices. AB - Three types of covalently crosslinked assemblies consisting of multiple (1) molecular layers of human serum albumin (HSA); (2) alternating layers of HSA and unfractionated heparin; and (3) alternating layers of HSA and partly depolymerized heparin fixed with one end to HSA were prepared on various surfaces. Adsorption of fibrinogen, IgG, and antithrombin (ATIII) from human citrated plasma on coated surfaces was evaluated by ELISA. Fibrinogen adsorption on coated ELISA plates was lower than that on bare polystyrene. There was no IgG adsorption on the HSA coating alone, but considerably high IgG adsorption was detected on the heparin-containing surface. The adsorption of ATIII increased with increasing heparin on the surface. The effect of multilayer coatings on platelets was tested by incubation of modified vascular prostheses with citrated blood. The most favorable interaction with platelets was observed on the HSA assembly. The interaction of platelets with the surface bearing unfractionated heparin was higher than that of the surface covered with partly depolymerized heparin. The long-term durability of the HSA-heparin coating was proven by a 21 day implantation of coated polyurethane plates in goat heart. PMID- 10825226 TI - Bioactive polymethyl methacrylate-based bone cement: comparison of glass beads, apatite- and wollastonite-containing glass-ceramic, and hydroxyapatite fillers on mechanical and biological properties. AB - A new bioactive bone cement (designated GBC) consisting of polymethyl methacrylate (PMMA) as an organic matrix and bioactive glass beads as an inorganic filler has been developed. The bioactive beads, consisting of MgO-CaO SiO(2)-P(2)O(5)-CaF(2) glass, have been newly designed, and a novel PMMA powder was selected. The purpose of the present study was to compare this new bone cement GBC's mechanical properties in vitro and its osteoconductivity in vivo with cements consisting of the same matrix as GBC and either apatite- and wollastonite-containing glass-ceramic (AW-GC) powder (designated AWC) or sintered hydroxyapatite (HA) powder (HAC). Each filler added to the cements amounted to 70 wt %. The bending strength of GBC was significantly higher than that of AWC and HAC (p < 0.0001). Cements were packed into intramedullar canals of rat tibiae in order to evaluate osteoconductivity as determined by an affinity index. Rats were sacrificed at 2, 4, and 8 weeks after operation. An affinity index, which equaled the length of bone in direct contact with the cement expressed as a percentage of the total length of the cement surface, was calculated for each cement. At each time interval studied, GBC showed a significantly higher affinity index than AWC or HAC up to 8 weeks after implantation (p < 0.03). The value for GBC increased significantly with time up to 8 weeks (p < 0.006). The handling property of GBC was comparable with that of PMMA bone cement. Our study revealed that the higher osteoconductivity of GBC was due to the higher bioactivity of the bioactive glass beads at the cement surface and the lower solubility of the new PMMA powder to MMA monomer. In addition, it was found that the smaller spherical shape and glassy phase of the glass beads gave GBC strong enough mechanical properties to be useful under weight-bearing conditions. GBC shows promise as an alternative with improved properties to the conventionally used PMMA bone cement. PMID- 10825227 TI - A biodegradable hybrid sponge nested with collagen microsponges. AB - A biodegradable hybrid sponge of poly(DL-lactic-co-glycolic acid) (PLGA) and collagen was fabricated by forming microsponges of collagen in the pores of PLGA sponge. Observation of the PLGA-collagen hybrid sponge by scanning electron microscopy (SEM) showed that microsponges of collagen with interconnected pore structures were formed in the pores of PLGA sponge. The hybrid structure further was confirmed by scanning electron microscopy-electron probe microanalysis (SEM EPMA), and elemental nitrogen was detected in the microsponges of collagen and on the pore surfaces of PLGA, but not in cross-sections of PLGA regions. The formation of collagen microsponges was dependent on collagen concentration, the effective range of which was from 0.1 to 1.5 (w/v) %. The mechanical strength of the hybrid sponge was higher than that of either PLGA or collagen sponges, in both dry and wet states. The wettability with water was improved by hybridization with collagen, which facilitated cell seeding in the hybrid sponge. Mouse fibroblast L929 cells attached well and spread on the surfaces of the microsponges of collagen in the hybrid sponge. The distribution of cells was spatially uniform throughout the hybrid sponge. Use of the PLGA sponge as a skeleton facilitated formation of the hybrid sponge into desired shapes with high mechanical strength while collagen microsponges contributed good cell interaction and hydrophilicity. PMID- 10825228 TI - A new approach to demonstrate cellular activity in bone formation adjacent to implants. AB - Bone tissue repeatedly formed in titanium 6-aluminum 4-vanadium rabbit bone harvest implants was collected in vivo at various times between 12 days and 5 weeks. Qualitative and quantitative examinations on undecalcified thin sections were performed in the light microscope. The amount of bone tissue was calculated on routinely stained sections. Alkaline (ALP) and acid phosphatase (ACP) enzyme activities were investigated. We also performed immunohistological detection of bone matrix proteins. Increasing bone density as well as an increasing mineralization of the tissue was observed in the biopsies with increasing time. The ALP and ACP activities were similar at short times (12 days and 2 weeks). With increasing time the ALP activity was stronger than that of ACP. The results from the immunohistochemical detection of osteonectin, osteopontin, bone sialoprotein, and collagen I and II demonstrated changes in the tissue differentiation with time. The tissue formation in the canal became more mature with time of ingrowth, as observed with the various techniques used in this study. Owing to these methodical developments, undecalcified ground sections may be used for detailed analysis of various phases of tissue formation in close proximity to implants. PMID- 10825229 TI - Histone deacetylases, transcriptional control, and cancer. AB - A key event in the regulation of eukaryotic gene expression is the posttranslational modification of nucleosomal histones, which converts regions of chromosomes into transcriptionally active or inactive chromatin. The most well studied posttranslational modification of histones is the acetylation of epsilon amino groups on conserved lysine residues in the histones' amino-terminal tail domains. Significant advances have been made in the past few years toward the identification of histone acetyltransferases and histone deacetylases. Currently, there are over a dozen cloned histone acetyltransferases and at least eight cloned human histone deacetylases. Interestingly, many histone deacetylases can function as transcriptional corepressors and, often, they are present in multi subunit complexes. More intriguing, at least some histone deacetylases are associated with chromatin-remodeling machines. In addition, several studies have pointed to the possible involvement of histone deacetylases in human cancer. The availability of the cloned histone deacetylase genes has provided swift progress in the understanding of the mechanisms of deacetylases, their role in transcription, and their possible role in health and disease. PMID- 10825230 TI - Cell-cycle-dependent turnover of P-glycoprotein in multidrug-resistant cells. AB - Regulation of P-glycoprotein (Pgp) expression occurs not only at the DNA and mRNA level but also at the protein level. We showed previously that Pgp was stabilized when multidrug-resistant CH(R)C5 and SKVCR 2.0 ovarian cell lines were subjected to serum-starved or high-cell-density growth conditions, whereas Pgp turnover in a leukemic multidrug-resistant cell line, CEMVLB0.1, was not affected by serum starvation (Muller et al., 1995). On further analysis, we have observed that the majority of the CH(R)C5 and SKVCR 2.0 cells under these conditions were in the G1/G0 phase of the cell cycle, whereas the cell cycle of CEMVLB0.1 cells was not affected. Pgp in CEMVLB0.1 cells was stabilized only when the cell cycle was delayed in the G1/G0 phase by using amino acid-deficient growth medium. In CH(R)C5 cells, Pgp half-life was also considerably increased when the cell cycle of these ovary-derived cells was delayed in the G1/G0 phase by using high concentrations of progesterone under normal serum growth conditions. In contrast, Pgp stability was not greatly affected if these cells were delayed in the S or G2/M phase of the cell cycle with Ara-C, cisplatin, or colchicine under the same conditions. Insulin-like growth factors could release the serum-starved CH(R)C5 and SKVCR2.0 cells from the G1/G0 phase and destabilized Pgp. These results indicate that Pgp turnover is a cell-cycle-related process in MDR cells. PMID- 10825231 TI - Effects of pertussis toxin on extracellular signal-regulated kinase activation in hepatocytes by hormones and receptor-independent agents: evidence suggesting a stimulatory role of G(i) proteins at a level distal to receptor coupling. AB - It was previously found that pertussis toxin (PTX) pretreatment inhibits the activation of extracellular signal-regulated kinases ERK1 (p44(mapk)) and ERK2 (p42(mapk)) in hepatocytes in response to either agonists that bind to heptahelical receptors or epidermal growth factor (EGF), suggesting a role of G(i) proteins in stimulatory mechanisms for ERK1/2. The present work shows that ERK1/2 is activated in a PTX-sensitive way not only by vasopressin, angiotensin II, prostaglandin (PG) F(2alpha), alpha(1)-adrenergic stimulation, and EGF but also by agents whose actions bypass receptors and stimulate protein kinase C (PKC) and/or elevate intracellular Ca(2+), such as 12-O-tetradecanoyl phorbol-13 acetate (TPA), exogenous phosphatidylcholine-specific phospholipase C (PC-PLC, from Bacillus cereus), thapsigargin, and the Ca(2+) ionophore A23187. Under the same conditions, PTX did not affect agonist stimulation of phosphoinositide specific phospholipase C (PI-PLC) (IP(3) generation), and did not reduce the activation by these agents of phospholipase D (PLD). The results suggest that in hepatocytes a PTX-sensitive mechanism, presumably involving G(i) proteins, exerts a stimulatory effect on ERK at a level distal to receptor coupling, acting either as an integral part of the signaling pathway(s) or by a permissive, synergistic regulation. PMID- 10825232 TI - Bone morphogenetic protein-7 (osteogenic protein-1) inhibits smooth muscle cell proliferation and stimulates the expression of markers that are characteristic of SMC phenotype in vitro. AB - Vascular proliferative disorders are characterized by migration and proliferation of vascular smooth muscle cells (SMCs), loss of expression of SMC phenotype, and enhanced extracellular matrix synthesis (e.g., type I collagen). We report here that bone morphogenetic protein-7 (BMP-7), a member of the transforming growth factor-beta (TGF-beta) superfamily, is capable of inhibiting both serum stimulated and growth factor-induced (platelet-derived growth factor [PDGF-BB] and TGF-beta1) cell growth as measured by (3)H-thymidine uptake into DNA synthesis and cell number in primary human aortic smooth muscle (HASM) cell cultures. Concomitantly, addition of BMP-7 stimulates the expression of SMC specific markers, namely alpha-actin and heavy chain myosin as examined by RT-PCR and Northern blot analyses. The collagen type III/I ratio that becomes lower with the transdifferentiation of SMCs into myofibroblasts is also maintained in BMP-7 treated cultures as compared to untreated controls. Studies on the mechanism of action indicate that BMP-7 treatment inhibits cyclin-dependent kinase 2 (cdk-2) that was stimulated during PDGF-BB-induced proliferation of SMCs and upregulates the expression of the inhibitory Smad, Smad6, which was shown to inhibit TGF-beta superfamily signaling. These results collectively suggest that BMP-7 maintains the expression of vascular SMC phenotype and may prevent vascular proliferative disorders, thus potentially acting as a palliative after damage to the vascular integrity. PMID- 10825233 TI - Synergistic activation of NF-kappab and inducible isoform of nitric oxide synthase induction by interferon-gamma and tumor necrosis factor-alpha in INS-1 cells. AB - Interferon-gamma (IFN-gamma) is known to exert deleterious effects on pancreatic beta-cells and is implicated in the development of type 1 (autoimmune) diabetes mellitus. In this study, we investigated signaling mechanisms mediating the effects of IFN-gamma in pancreatic beta-cells using a differentiated rat insulin secreting cell line, INS-1, with special reference to the activation of transcription factors STAT (signal transducers and activators of transcription)1 and NF-kappaB. Exposure of INS-1 cells to 100 IU/ml IFN-gamma for 24 h resulted in significant inhibition of nutrient-induced insulin secretion associated with impaired metabolism. In combination with tumor necrosis factor-alpha (TNF-alpha) (50 ng/ml), IFN-gamma elicited severe cytotoxicity and induced the expression of the inducible isoform of nitric oxide synthase (iNOS) mRNA. IFN-gamma promoted tyrosine phosphorylation and DNA-binding of STAT1 through Janus kinase (JAK)1 activation without apparent phosphorylation of JAK2. TNF-alpha did not affect STAT1 activation, but stimulated DNA-binding and transcriptional activity of NF kappaB, both of which were further increased by IFN-gamma. These effects of IFN gamma and TNF-alpha seem physiologically relevant, because either inhibition of STAT1 by the tyrosine kinase inhibitor herbimycin A or that of NF-kappaB by sulfasalazine resulted in the reduction of iNOS mRNA expression. In conclusion, IFN-gamma activates STAT1 and potentiates TNF-alpha-induced NF-kappaB activation in INS-1 cells, thereby inducing iNOS and cell destruction. PMID- 10825234 TI - Human mesenchymal stem cells support megakaryocyte and pro-platelet formation from CD34(+) hematopoietic progenitor cells. AB - Megakaryocytopoiesis and thrombocytopoiesis result from the interactions between hematopoietic progenitor cells, humoral factors, and marrow stromal cells derived from mesenchymal stem cells (MSCs) or MSCs directly. MSCs are self-renewing marrow cells that provide progenitors for osteoblasts, adipocytes, chondrocytes, myocytes, and marrow stromal cells. MSCs are isolated from bone marrow aspirates and are expanded in adherent cell culture using an optimized media preparation. Culture-expanded human MSCs (hMSCs) express a variety of hematopoietic cytokines and growth factors and maintain long-term culture-initiating cells in long-term marrow culture with CD34(+) hematopoietic progenitor cells. Two lines of evidence suggest that hMSCs function in megakaryocyte development. First, hMSCs express messenger RNA for thrombopoietin, a primary regulator for megakaryocytopoiesis and thrombocytopoiesis. Second, adherent hMSC colonies in primary culture are often associated with hematopoietic cell clusters containing CD41(+) megakaryocytes. The physical association between hMSCs and megakaryocytes in marrow was confirmed by experiments in which hMSCs were copurified by immunoselection using an anti-CD41 antibody. To determine whether hMSCs can support megakaryocyte and platelet formation in vitro, we established a coculture system of hMSCs and CD34(+) cells in serum-free media without exogenous cytokines. These cocultures produced clusters of hematopoietic cells atop adherent MSCs. After 7 days, CD41(+) megakaryocyte clusters and pro-platelet networks were observed with pro-platelets increasing in the next 2 weeks. CD41(+) platelets were found in culture medium and expressed CD62P after thrombin treatment. These results suggest that MSCs residing within the megakaryocytic microenvironment in bone marrow provide key signals to stimulate megakaryocyte and platelet production from CD34(+) hematopoietic cells. PMID- 10825235 TI - Generation of active TGF-beta by prostatic cell cocultures using novel basal and luminal prostatic epithelial cell lines. AB - Two prostatic epithelial lines, one of basal origin and one of luminal origin, were established from the dorsolateral prostates of p53 null mice. The cell lines are nontumorigenic when inoculated subcutaneously under the renal capsule or intraprostatically in syngeneic mice. The luminal cell line (PE-L-1) expresses cytokeratins 8 and 18 and the basal cell line (PE-B-1) expresses cytokeratins 5 and 14. The basal cells require serum for growth, whereas the luminal cells grow only in serum-free medium. Both cell lines require the presence of growth factors for optimal growth in culture, with EGF and FGF-2 having the greatest effect on the growth rate. Both lines express androgen receptor (AR) mRNA and protein. Androgen stimulates growth of the basal cell line, indicating that the ARs are functional, whereas growth of the luminal cells is unaffected by androgens. The luminal line is significantly inhibited by exogenous TGF-beta and produces low levels of endogenous TGF-beta. In contrast, the basal cell line produces significant amounts of TGF-beta and its growth is not influenced by this cytokine. Coculture of luminal cells with prostatic smooth muscle cells results in the generation of increased levels of biologically active TGF-beta, indicating a paracrine mechanism of TGF-beta activation that may be involved in the maintenance of normal prostatic function. To our knowledge this is the first report describing both basal and luminal prostatic cell lines from a single inbred animal species and the first indication that prostatic epithelial and stromal cells interact to generate the biologically active form of TGF-beta. These lines will provide an important model for determining basal/luminal interactions in both in vitro and in vivo assays. PMID- 10825236 TI - p21(cip1) mRNA is controlled by endogenous transforming growth factor-beta1 in quiescent human hematopoietic stem/progenitor cells. AB - Transforming growth factor-beta1 (TGF-beta1) has been described as an efficient growth inhibitor that maintains the CD34(+) hematopoietic progenitor cells in quiescence. The concept of high proliferative potential-quiescent cells or HPP-Q cells has been introduced as a working model to study the effect of TGF-beta1 in maintaining the reversible quiescence of the more primitive hematopoietic stem cell compartment. HPP-Q cells are primitive quiescent stem/progenitor cells on which TGF-beta1 has downmodulated the cytokine receptors. These cells can be released from quiescence by neutralization of autocrine or endogenous TGF-beta1 with a TGF-beta1 blocking antibody or a TGF-beta1 antisense oligonucleotide. In nonhematopoietic systems, TGF-beta1 cooperates with the cyclin-dependent kinase inhibitor, p21(cip1), to induce cell cycle arrest. We therefore analyzed whether endogenous TGF-beta1 controls the expression of the p21(cip1) in the CD34(+) undifferentiated cells using a sensitive in situ hybridization method. We observed that addition of anti-TGF-beta1 is followed by a rapid decrease in the level of p21(cip1) mRNA whereas TGF-beta1 enhances p21(cip1) mRNA expression concurrently with an inhibitory effect on progenitor cell proliferation. These results suggest the involvement of p21(cip1) in the cell cycle control of early human hematopoietic quiescent stem/progenitors and not only in the differentiation of more mature myeloid cells as previously described. The modulation of p21(cip1) observed in response to TGF-beta1 allows us to further precise the working model of high proliferative potential-quiescent cells. PMID- 10825237 TI - Cell coupling modulates the contraction of fibroblast-populated collagen lattices. AB - Cultured dermal fibroblasts become notably elongated when incorporated into a fibroblast-populated collagen lattice (FPCL). With time these fibroblasts reorganize the collagen responsible for reduction in lattice size. In monolayer the microinjection of Lucifer Yellow (LY) into cultured human fibroblasts shows cell coupling through gap junctions. Human fibroblasts residing on the periphery of a FPCL are at high density and the microinjection of LY into one of those fibroblasts demonstrates cell coupling. Cells within the center of an FPCL are at low density and appear to be independent of one another; however, the microinjection of LY into selected fibroblasts again demonstrates cell coupling. Hence the microinjection of cells in both the center and the edge of a FPCL pass dye to numerous neighbors. Does cell coupling influence FPCL contraction? FPCL incubated with heptanol and octanol, aliphatic alcohols that uncouple cells, inhibits lattice contraction, whereas hexanol, an aliphatic alcohol that does not uncouple cells, did not alter lattice contraction. Fibroblasts derived from connexin 43 (a transmembrane protein responsible for gap junction structures) knockout mice were demonstrated to lack gap junctional communications. When incorporated into a FPCL these cells failed to elongate and demonstrated retarded lattice contraction. Hence, gap junctional communications between fibroblasts incorporated into collagen lattices appear to optimize FPCL contraction and suggest a role for gap junctions in the organization of collagen fibers. PMID- 10825238 TI - Endothelial heparan sulfate is necessary but not sufficient for control of vascular smooth muscle cell growth. AB - The state of the endothelial cell (EC) determines the nature of its control of vascular smooth muscle cell (vSMC) biology. Conditioned medium from postconfluent ECs inhibits vSMC proliferation, whereas subconfluent conditioned medium from the same ECs has a stimulatory effect. We and others have identified confluent endothelial cells' production of heparan sulfate proteoglycans (HSPG) as critical to vSMC growth control. The question that arises is whether the stimulation that is observed with subconfluent cells is from (1) aberrant HSPG production, (2) elaboration of noninhibitory species of HSPG, or (3) production of other factors, such as mitogens, which counteract the inhibitory HSPG to stimulate vSMCs. We studied the relative effects of conditioned medium produced by both subconfluent and postconfluent EC cultures on vSMC growth. Conditioned medium was fractionated into nonproteoglycan (non-PG) and proteoglycan (PG) components by anion-exchange chromatography. The PG fractionation profile and the antiproliferative activity of the HSPGs isolated from both subconfluent and postconfluent EC-conditioned media were similar. However, the HSPG fraction alone could not approach the inhibitory potential of unfractionated conditioned medium from postconfluent EC cultures. Non-PG proteins produced by the endothelial cultures had no effect on vSMC growth on their own. Yet, when they were mixed together with HSPG fractions, from either subconfluent or postconfluent EC cultures, the full growth effects were returned. Non-PG protein fractions from postconfluent cultures with HSPG fractions gave maximal inhibition of vSMC growth, whereas non-PG protein fractions from subconfluent EC cultures with HSPG fractions produced the maximal stimulation. Thus, whereas the net stimulatory or inhibitory effect on vSMC growth of EC-conditioned medium is density dependent, this effect does not result from a difference in the antiproliferative heparan sulfate component but rather from non-PG proteins that interact with the heparan sulfates. PMID- 10825239 TI - Hepatocyte growth factor (HGF) inhibits skeletal muscle cell differentiation: a role for the bHLH protein twist and the cdk inhibitor p27. AB - Hepatocyte growth factor (HGF) plays a crucial role in regulating the differentiation of both fetal and adult skeletal myoblasts. This study aimed at defining the intracellular factors that mediate the effect of HGF on adult myoblast differentiation. HGF increased Twist expression while decreasing p27(kip1) protein levels and not affecting the induction of p21(Cip1/Waf1) in satellite cells. Like HGF, overexpression of Twist did not affect p21 expression while inhibiting muscle-specific proteins. Both ectopic Twist-antisense (Twist AS) and p27 partially rescued the effects of HGF on bromodeoxyuridine (BrdU) incorporation and myosin heavy chain (MHC) expression in muscle satellite cells; the two plasmids together effected full rescue, suggesting that HGF independently regulates these two factors to mediate its effects. Ectopic p27 promoted differentiation in the presence of HGF by blocking the induction of Twist. Using Twist-AS to lower Twist levels restored the HGF-dependent reduction of p27 and MHC. In the presence of ectopic HGF, satellite cells formed thin mononuclear myotubes. Neither ectopic p27, Twist-AS, or their combination reversed this change in cell morphology, suggesting that HGF acts through additional mediators to inhibit downstream events during myogenesis. Taken together, the results suggest that the effects of HGF on muscle cell proliferation and differentiation are mediated through changes in the expression levels of the myogenic-inhibitory basic helix-loop-helix (bHLH) protein Twist and the cell-cycle inhibitor p27. PMID- 10825240 TI - Gap junctions in human synovial cells and tissue. AB - Our objective was to establish the existence of intercellular communication through gap junctions in synovial lining cells and in primary and passaged cultures of human synovial cells. Communication between cells was assessed using the nystatin perforated-patch method, fluorescent dye transfer, immunochemistry, transmission electron microscopy, and immunoblotting. Functional gap junctions were observed in primary and passaged cultures and were based on measurements of the transient current response to a step voltage. The average resistance between cells in small aggregates was 300 +/- 150 MOmega. Gap junctions were also observed between synovial lining cells in tissue explants; the size of the cell network in synovial tissue was estimated to be greater than 40 cells. Intercellular communication between cultured cells and between synovial lining cells was confirmed by dye injection. Punctate fluorescent regions were seen along intercellular contacts between cultured cells and in synovial membranes in cells and tissue immunostained for connexin43. The presence of the protein was verified in immunoblots. Regular 2-nm intermembrane gap separations characteristic of gap junctions were seen in transmission electron micrographs of synovial biopsies. The results showed that formation of gap-junction channels capable of mediating ionic and molecular communication was a regular feature of synovial cells, both in tissue and in cultured cells. The gap junctions contained connexin43 protein and perhaps other proteins. The physiological purpose of gap junctions in synovial cells is unknown, but it is reasonable to anticipate that intercellular communication serves some presently unrecognized function. PMID- 10825241 TI - Intracellular osteopontin is an integral component of the CD44-ERM complex involved in cell migration. AB - Osteopontin (OPN) is a secreted glycoprotein with mineral- and cell-binding properties that can regulate cell activities through integrin receptors. Previously, we identified an intracellular form of osteopontin with a perimembranous distribution in migrating fetal fibroblasts (Zohar et al., J Cell Physiol 170:88-98, 1997). Since OPN and CD44 expression are increased in migrating cells, we analyzed the relationship of these proteins with immunofluorescence and confocal microscopy. A distinct co-localization of perimembranous OPN and cell-surface CD44 was observed in fetal fibroblasts, periodontal ligament cells, activated macrophages, and metastatic breast cancer cells. The co-localization of OPN and CD44 was prominent at the leading edge of migrating fibroblasts, where OPN also co-localized with the ezrin/radixin/moesin (ERM) protein ezrin, as well as in cell processes and at attachment sites of hyaluronan-coated beads. The subcortical location of OPN in these cells was verified by cell-surface biotinylation experiments in which biotinylated CD44 and non-biotinylated OPN were isolated from complexes formed with hyaluronan-coated beads and identified with immunoblotting. That perimembranous OPN represents secreted protein internalized by endocytosis or phagocytosis appeared to be unlikely since exogenous OPN that was added to cell cultures could not be detected inside the cells. A physical association with OPN, CD44, and ERM, but not with vinculin or alpha-actin, was indicated by immunoadsorption and immunoblotting of cell proteins in complexes extracted from hyaluronan-coated beads. The functional significance of OPN in this complex was demonstrated using OPN-/- and CD-/- mouse fibroblasts which displayed impaired migration and a reduced attachment to hyaluronan-coated beads. These studies indicate that OPN exists as an integral component of a hyaluronan-CD44-ERM attachment complex that is involved in the migration of embryonic fibroblasts, activated macrophages, and metastatic cells. PMID- 10825242 TI - Interruption of NFkappaB-STAT1 signaling mediates EGF-induced cell-cycle arrest. AB - It is known that EGF induces the cell-cycle arrest in A431 cells that possess high numbers of EGF receptors and it was previously suggested that p21/WAF1 protein was a major effector molecule of the EGF-mediated cell-cycle arrest of A431 cells. Here, we further investigate this phenomenon using the decoy double strand oligonucleotides for STAT-binding sequence (STAT decoy) and IkappaB, an inhibitor of the nuclear factor kappa B (NFkappaB). Addition of STAT decoy restored EGF-induced A431 cell-growth arrest. Interestingly, infection of adenovirus vectors to express IkappaB (AxIkappaBalphaDeltaN) as the inhibitor of NFkappaB also reversed the A431 cell-growth inhibition. The individual treatment of two inhibitors partially inhibited the WAF1 gene expression, whereas simultaneous treatment of two inhibitors exhibited more efficient inhibition. These observations suggest the activation of NFkappaB via IkappaB degradation and STAT1 via specific receptor kinase activity synergistically induce WAF1 gene expression in A431 cells. Thus, NFkappaB and STAT1 pathways mutually interact to play an important role in the EGF-induced intracellular reaction. PMID- 10825243 TI - 252Cf plasma desorption mass spectrometric study of interactions of steroid glycosides with amino acids AB - 252Cf Plasma desorption with time-of-flight mass spectrometry (TOF-PDMS) has been applied to comparative studies of the interactions of steroid glycosides (SGs) of the spirostan series with amino acids. SGs can interact with amino acids to form heteroclusters of the type [SG + aminoacid + H](+) and [SG + aminoacid + K](+). It is shown how the affinity of SGs for amino acids depends on the structures of both the SG carbohydrate chain and the SG aglycone, and on the nature of the amino acid side chain. Copyright 2000 John Wiley & Sons, Ltd. PMID- 10825244 TI - Liquid chromatography/electrospray ionisation sequential mass spectrometric identification of the main chlortoluron by-products during water disinfection using chlorine AB - The main degradation by-products of the herbicide chlortoluron formed during water disinfection with HOCl/ClO(-) have been separated and identified by liquid chromatography/electrospray ionisation sequential mass spectrometry (LC/ESI MS(n)). Chlorination and hydroxylation reactions seem to occur exclusively on the aromatic ring of chlortoluron, leading to by-products which show characteristic fragmentation patterns. Indeed, chlortoluron-like ESI spectra were always observed for chlorinated by-products, showing only a fragment at m/z 72. In contrast, hydroxylated and chloro/hydroxylated by-products gave a much more complex fragmentation pattern that could be elucidated by MS(n) (n = 1-4) experiments. A mechanistic scheme rationalising the observed fragmentation pattern is proposed and discussed. Copyright 2000 John Wiley & Sons, Ltd. PMID- 10825245 TI - Application of matrix-assisted laser desorption/ionization to on-line aerosol time-of-flight mass spectrometry AB - Matrix-assisted laser desorption/ionization (MALDI) mass spectra were obtained from single biological aerosol particles using an aerosol time-of-flight mass spectrometer (ATOFMS). The inlet to the ATOFMS was coupled with an evaporation/condensation flow cell that allowed the aerosol to be coated with matrix material as the sampled stream entered the spectrometer. Mass spectra were generated from aerosol composed either of gramicidin-S or erythromycin, two small biological molecules, or from aerosolised spores of Bacillus subtilis var niger. Three different matrices were used: 3-nitrobenzyl alcohol, picolinic acid and sinapinic acid. A spectrum of gramicidin-S was generated from approximately 250 attomoles of material using a molar ratio of 3-nitrobenzyl alcohol to analyte of approximately 20:1. A single peak, located at 1224 Da, was obtained from the bacterial spores. The washing liquid and extract solution from the spores were analyzed using electrospray mass spectrometry and subsequent MS/MS product ion experiments. This independent analysis suggests that the measured species represents part of the B. subtilis peptidoglycan. The on-line addition of matrix allows quasi-real-time chemical analysis of individual, aerodynamically sized particles, with an overall system residence time of less than 5 seconds. These results suggest that a MALDI-ATOFMS can provide nearly real-time identification of biological aerosols. Copyright 2000 John Wiley & Sons, Ltd. PMID- 10825246 TI - Identification and characterization of fenton oxidation products of surfactants by electrospray mass spectrometry and by solid phase microextraction gas chromatography mass spectrometry. 2. Fatty alcohol polyethoxy sulphates AB - The Fenton reaction for the degradation of surfactants has been investigated and partial degradation products have been identified and characterized by mass spectrometry for the case of fatty alcohol polyethoxy sulphates. The polar water soluble products were investigated by liquid chromatography/mass spectrometry (LC/MS) with electrospray ionization (ESI), and the volatile products leaving the mixture during the reaction were trapped by means of solid phase microextraction (SPME) and investigated by gas chromatography/mass spectrometry (GC/MS) with electron and chemical ionization. The oxidation leads to the formation of products with hydroxyl and epoxide groups due to insertion of oxygen atoms or with terminal ethoxylic moieties deriving from the loss of the hydrophilic sulphate group. The formation of volatile aldehydes is also observed, corresponding to the fragmentation between hydrocarbon and ethoxylic chains. The extent of mineralization is dependent on peroxide and iron(II) concentrations. Copyright 2000 John Wiley & Sons, Ltd. PMID- 10825247 TI - Probing acrylamide alkylation sites in cysteine-free proteins by matrix-assisted laser desorption/ionisation time-of-flight. AB - It is recognised that gel-separated proteins can experience a frequent modification provoked by the interaction of unpolymerized acrylamide monomers with the thiol group of cysteine to form a beta-cysteinyl-S-propionamide adduct. Other groups which have been implicated in this reaction include the hydroxyl group of tyrosine, the straightepsilon-amino group of lysine, and the free N terminus. In a series of recent publications it has been demonstrated that at pH approximately 9.5 and in the presence of cysteine, none of these groups experienced measurable interaction with acrylamide monomers. To emphasise this conclusion we have used matrix-assisted laser desorption/ionisation with a reflectron time-of-flight mass spectrometer to examine a number of cysteine-free proteins incubated for various intervals with 30 mM acrylamide monomers at pH 9.5. These high resolution data suggest that, for short incubation times (>/=1 hour) and in the absence of cysteine, the straightepsilon-NH(2) group of lysine is the likely adduction site of acrylamide. Longer incubation times (>/=24 hours) with acrylamide monomers rendered the role of Cys as the favourite alkylation site less evident. PMID- 10825248 TI - Derivatization for electrospray ionization-mass spectrometry. 4. Alkenes and alkynes AB - A survey of derivatization strategies and prospective derivatization reactions for conversion of simple alkenes and alkynes to 'electrospray-active' species is presented. General synthetic strategies are discussed and illustrative examples of prospective derivatives prepared from model compounds are presented along with their electrospray ionization (ES) mass spectra. The identified derivatives of these neutral, nonpolar analytes are either ionic or are ionizable in solution through Bronsted acid/base chemistry, by Lewis acid/base chemistry, or by chemical or electrochemical electron-transfer chemistry. Once ionized, the derivatives are expected to be amenable to detection by electrospray ionization mass spectrometry. Derivatives are identified for positive and negative ion analysis of both alkenes and alkynes. Copyright 2000 John Wiley & Sons, Ltd. PMID- 10825249 TI - Detection of intact hemoglobin from aqueous solution with laser desorption mass spectrometry. AB - Laser induced liquid beam ionization/desorption mass spectrometry (LILBID-MS) is a new desorption method recently developed in our laboratory. This method allows ions to be desorbed directly from the liquid phase into the high-vacuum region of a mass spectrometer. This method has now been applied to the detection of noncovalent protein-protein complexes. The example given in this paper is the quartenary complex of human hemoglobin. For the first time, the intact hemoglobin could be detected by laser desorption mass spectrometry. Furthermore, evidence for the specificity of the complex is given. PMID- 10825250 TI - N-(2-diethylamino)ethyl-4-aminobenzamide derivative for high sensitivity mass spectrometric detection and structure determination of N-linked carbohydrates. AB - N-Linked glycans were derivatised by reductive amination using N-(2 diethylamino)ethyl-4-aminobenzamide (DEAEAB, procainamide) and examined by electrospray mass spectrometry. This derivative ionised primarily by protonation of the tertiary amine group and attachment of an alkali metal to give [M + H + X](2+) ions which were much more abundant that doubly charged ions from glycans derivatised with other aromatic amines. Fragmentation of these ions depended on the nature of the alkali metal (X). Lithium and sodium adducts fragmented to give prominent ions produced by cleavages within the DEAEAB derivative whereas the other adducts produced more abundant ions from fragmentation of the carbohydrate. Elimination of a sugar fragment, usually by cleavage adjacent to GlcNAc or sialic acid, together with a hydrogen atom, produced the most abundant singly charged fragment ions. These ions then formally fragmented by glycosidic cleavages. Potassium, rubidium and caesium adducts produced abundant losses of the alkali metal, but the resulting ions appeared not to undergo extensive fragmentation. Most fragment ions from all of the adducts were singly charged, the remainder being doubly charged. Although the spectra of the [M + X + H](2+) ions were not as informative as those from the singly charged ions from other derivatives, they, nevertheless, provided much valuable information on the structure of these glycans. PMID- 10825251 TI - Ion/molecule reactions with dimethyl ether and dimethyl-d6 ether: differentiation of four isomers contained in patchouli essential oil AB - In this study, we show that it is possible to differentiate four sesquiterpene isomers (C(15)H(24)) preliminarily separated by gas chromatography/mass spectrometry (GC/MS). Dimethyl ether is evaluated as a selective ionization reagent and the relative abundance of adducts formed with this reagent gas under positive chemical ionization conditions are compared and adduct ions are characterized using collision-induced dissociation. The mechanisms have been confirmed by achieving the same experiments with deuterated dimethyl ether. Copyright 2000 John Wiley & Sons, Ltd. PMID- 10825252 TI - Investigation of unknown related substances in commercial erythromycin samples with liquid chromatography/mass spectrometry. AB - A selective reversed phase liquid chromatography/mass spectrometry (LC/MS(n)) method is described for the identification of erythromycin impurities and related substances in commercial erythromycin samples. Mass spectral data are acquired on a LCQ ion trap mass spectrometer equipped with an electrospray interface operated in positive ion mode. The LCQ is ideally suited for identification of impurities and related substances because it provides on-line LC/MS(n) capability. Compared with UV detection, this hyphenated LC/MS(n) technique provides as a main advantage efficient identification of novel substances without time-consuming isolation and purification procedures. Using this method four novel related substances were identified in commercial samples. PMID- 10825253 TI - Effect of alkali metal cationization and multiple alkali metal exchange on the collision-induced dissociation of loganic acid studied by electrospray ionization tandem mass spectrometry AB - Under electrospray ionization conditions loganic acid undergoes alkali metal (Li, Na and K) exchange and alkali metal cationization. Multiple exchanges of up to four alkali metal ions are observed. Different populations of metal exchanged species are produced during electrospray ionization. Collision-induced dissociation of ammonium cationized species is compared with that of metal cationized species to study the effect of metal cationization. Glycosidic cleavage and ring cleavages of aglycone and sugar moieties are the major fragmentation pathways observed during collision-induced dissociation. The fragmentations of the highly metal exchanged species indicate the opening of the pyran ring. Collision-induced dissociation of the various metal exchanged and metal cationized species also reveals the nature of the different populations. Copyright 2000 John Wiley & Sons, Ltd. PMID- 10825254 TI - Qualitative and quantitative analysis of wheat gluten proteins by liquid chromatography and electrospray mass spectrometry. AB - Based on analysis by liquid chromatography/electrospray ionisation mass spectrometry, we have developed a new method for fast and sensitive fingerprinting of gliadins and glutenins in wheat flour. Using this procedure the two protein fractions from seven durum wheat varieties have been analysed by high resolution high performance liquid chromatographic separation coupled to accurate determination of molecular mass. In this way, the molecular mass of the single components from both gliadin and glutenin fractions were measured and more than forty components were detected for each fraction indicating a high heterogeneity. Although the chromatographic profiles were similar, the molecular masses of protein components with similar retention times among the varieties were often different. The difference ranged from a few mass units corresponding to single amino acid substitution(s) up to thousands implying peptide deletion or insertion along the protein chain. Two components representing about a half of the gliadin fraction, e.g. gamma(2)- and gamma(3)-gliadin, were identified through the N terminal sequence and molecular mass determination. We suggest the use of the high level and the molecular mass of these gliadin components as markers to detect traces of wheat in gluten-free food preparations for celiac patients. PMID- 10825256 TI - Matrix-assisted laser desorption/ionization time-of-flight mass spectrometric detection of bacterial biomarker proteins isolated from contaminated water, lettuce and cotton cloth. AB - Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectra of bacterial proteins were obtained from water, lettuce and cloth samples contaminated with Shigella flexneri, Escherichia coli, and Aeromonas hydrophila. Spectra were obtained using proteins directly isolated from water (or water used for rinsing samples) without culturing the bacteria. For S. flexneri and E. coli, two marker ions for specific proteins associated with a virulence-related property (acid resistance) were easily detected. For A. hydrophila, ions from two specifically selected marker proteins, as well as ions from the larger group of proteins isolated from pure cultures, all matched spectra from a contaminated water sample, providing strong evidence that A. hydrophila was the bacterial contaminant. Rinse water from contaminated lettuce and cloth samples showed the same marker ions as the contaminated water samples. PMID- 10825255 TI - Biotreatment of tannin-rich beer-factory wastewater with white-rot basidiomycete Coriolopsis gallica monitored by pyrolysis/gas chromatography/mass spectrometry. AB - Some fractions of beer-factory wastewaters represent an important environmental concern owing to their high content of polyphenols and dark-brown color. The capacity of Coriolopsis gallica to preferentially degrade lignin has been successfully applied in our laboratory to the biotreatment and decolorization of paper-industry effluents. In this work, the ability of this white-rot fungus to degrade high-tannin-containing wastewaters is evaluated. Under all the conditions studied, effluent decolorization and chemical oxygen demand reduction achieved by C. gallica at day 12 of incubation were close to 50 and 65%, respectively. No adhesion of dark color to the fungal mycelium was observed suggesting that decolorization could be ascribed to C. gallica degradation systems. Mycelium dry weight values showed that C. gallica is tolerant to relatively high tannin content present in the effluent samples. In the sample containing the highest effluent concentration (60% v/v), dry-weight values suggested an inhibition of fungal growth at day 6 of incubation and a further adaptation of the fungus to the stressing tannin effect at day 12 of fungal treatment. Pyrolysis/gas chromatography/mass spectrometry results showed a decrease of polyphenols pyrolysis products, mainly phenol and guaiacol, with the incubation time. All these results indicate the potential use of C. gallica in bioremediation of tannin-containing industrial wastewaters and in other applications where a reduction in polyphenols content is required. PMID- 10825257 TI - Identification of the protein-drug adduct formed between aldehyde dehydrogenase and S-methyl-N,N-diethylthiocarbamoyl sulfoxide by on-line proteolytic digestion high performance liquid chromatography electrospray ionization mass spectrometry. AB - Disulfiram has been used clinically as an aversion therapy treatment for recovering alcoholics. One of its metabolites, S-methyl-N, N-diethylthiocarbamoyl sulfoxide (MeDTC-SO), is currently believed by some to be the active metabolite in vivo. We demonstrate in this report that MeDTC-SO is a potent irreversible inhibitor of recombinant rat liver mitochondrial aldehyde dehydrogenase (rlmALDH), the enzyme responsible for oxidizing acetaldehyde formed during ethanol metabolism. Recombinant rlmALDH was inhibited by MeDTC-SO after in vitro incubation with an IC(50) = 4.62 microM. The inhibition of rlmALDH was found to be accompanied by a concomitant increase of approximately 100 Da to the molecular mass of the native enzyme as determined by on-line high performance liquid chromatography (HPLC) electrospray ionization mass spectrometry (LC/MS), indicating that a covalent modification has occurred. To determine the site and structure of this covalent adduct, we developed a novel approach to characterize specific protein-drug interactions by linking a proteolytic enzyme digestion cartridge on-line with LC/MS. The on-line pepsin digestion LC/MS of MeDTC-SO inhibited rlmALDH revealed an ion at MH(2)(2+) = 500.9, which was not present in the pepsin digestion of the non-inhibited enzyme. This peptide was tentatively attributed to the putative active site peptide (FNQGQC(301)C(302)C(303)) plus the adduct. This peptide was subjected to analysis by LC/MS/MS, which allowed us to determine that the covalent modification was associated with a single carbamoyl adduct at Cys-302, which has been shown to be the active site nucleophile of the enzyme. PMID- 10825258 TI - Sequencing of sulfonic acid derivatized peptides by electrospray mass spectrometry AB - We report the application of nanoelectrospray ionization tandem mass spectrometry (nES-MS/MS) and capillary LC/microelectrospray MS/MS (cLC/&mgr;ES-MS/MS) for sequencing sulfonic acid derivatized tryptic peptides. These derivatives were specifically prepared to facilitate low-energy charge-site-initiated fragmentation of C-terminal arginine-containing peptides, and to enhance the selective detection of a single series of y-type fragment ions. Both singly and doubly protonated peptides were analyzed by MS/MS and the results were compared with those from their derivatized counterparts. Model peptides and peptides from tryptic digests of gel-isolated proteins were analyzed. Derivatized singly protonated peptides fragment in the same way by nES-MS/MS as they do by post source decay matrix-assisted laser desorption/ionization mass spectrometry (PSD MALDI-MS). They produce fragment ion spectra dominated by y-ions, and the simplified spectra are readily interpreted de novo. Doubly protonated peptides fragment in much the same way as their non-derivatized doubly protonated counterparts. The fragmentation of doubly protonated derivatives is especially useful for sequencing peptides that possess a proline residue near the N-terminus of the molecule. The singly protonated forms of these proline-containing derivatives often show enhanced fragmentation on the N-terminal side of the proline and considerably reduced fragmentation on the C-terminal side. In addition, sulfonic acid derivatization increases the in-source fragmentation of arginine-containing peptides. This could be useful for sequence verification and sequence tagging for use in single stage mass spectrometry. Copyright 2000 John Wiley & Sons, Ltd. PMID- 10825259 TI - Rapid and sensitive determination of paclitaxel (Taxol(R)) in environmental samples by high-performance liquid chromatography tandem mass spectrometry AB - A new, rapid and sensitive method for the quantitative determination of paclitaxel (Taxol(R)) in environmental samples is developed and validated. This highly sensitive and specific method was successfully applied to an environmental monitoring study for identifying occupational exposure situations to anticancer drugs. After addition of an internal standard, 2'-methylpaclitaxel, aqueous extraction of wipe samples or air filters followed by a single-step liquid-liquid extraction with ethyl acetate was performed. Extraction efficiency averaged 90%. The method is based on analysis by liquid chromatography with tandem mass spectrometry in selected reaction monitoring mode of environmental samples from the preparation and administration of cytostatic agents. The assay is linear (r = 0.998) in the range 2-64 ng on filters (air samples), and 20-320 ng on wipe samples. The accuracy of the assay is always 0.10). However, increasing PaCO(2) from 41 to 58 mm Hg resulted in a 30% increase in CBV (3.08 +/- 0.86 mL/100 g, P < 0.05) and a 91% increase in CBF (97 +/- 39 mL/100 g/min, P < 0.01). In the control group, there were no significant changes in CBV and CBF (P > 0.10) during 2 h of propofol anesthesia. These results indicate that, during propofol anesthesia, cerebrovascular reactivity of blood flow and blood volume is maintained during hypercapnia but is markedly diminished during hypocapnia. IMPLICATIONS: During propofol anesthesia in rabbits with normal brains, a reduction in the arterial carbon dioxide level may not always be accompanied by a reduction in brain blood flow and blood volume. PMID- 10825325 TI - Ambulatory labor epidural analgesia: bupivacaine versus ropivacaine. AB - Dilute concentrations of bupivacaine combined with fentanyl have recently been used to initiate labor epidural analgesia in an attempt to balance adequate analgesia and minimal maternal motor blockade. Similar concentrations of ropivacaine have not been evaluated. This prospective, randomized, double-blinded study was designed to compare the efficacy of 20 mL of either 0.08% bupivacaine plus 2 microg/mL fentanyl or 0.08% ropivacaine plus 2 microg/mL fentanyl to initiate ambulatory labor epidural analgesia. Forty nulliparous women in early ( 0 but < 20 at 20 min. The time (mean +/- SD) to visual analog scale score = 0 was BF (n = 18): 12.0 +/- 4.5 min and RF (n = 19): 12.4 +/- 4.0 min (P > 0.05). Spontaneous micturition was observed in 65% (13 of 20) BF compared with 100% (20 of 20) RF (P < 0.01), and ambulation was demonstrated in 75% (15 of 20) BF compared with 100% (20 of 20) RF (P < 0.03). The incidence of forceps delivery was 35% (7 of 20) BF compared with 10% (2 of 20) RF (P < 0.04). The results of this study indicate that dilute ropivacaine combined with fentanyl effectively initiates epidural analgesia while concurrently preserving maternal ability to void and ambulate. IMPLICATIONS: As compared with a similar dilute concentration of bupivacaine, 20 mL of dilute (0.08%) ropivacaine combined with fentanyl (2 microg/mL) effectively initiates epidural analgesia in nulliparous women in early, established labor while preserving their ability to micturate and ambulate. Of importance, it appears that a true ambulatory epidural analgesic for women in labor is now possible. PMID- 10825326 TI - A dose-response study of prophylactic intravenous ephedrine for the prevention of hypotension during spinal anesthesia for cesarean delivery. AB - We performed a randomized, double-blinded dose-finding study of IV ephedrine for prophylaxis for hypotension in 80 women who received an IV crystalloid preload and spinal anesthesia for elective cesarean delivery. One minute after the intrathecal injection, patients were given saline control or ephedrine 10, 20, or 30 mg IV for 30 s. Systolic arterial pressure (SAP) in the first 12 min after the spinal injection was greater in the 30-mg group compared with other groups (P < 0.05). Hypotension occurred in 7 patients (35%) in the 30-mg group compared with 19 (95%), 17 (85%), and 16 (80%) patients in the control and 10- and 20-mg groups, respectively (P < 0.0001). Maximum decrease in SAP was smaller in the 30 mg group (mean lowest SAP 87% of baseline, range 58%-105%) compared with other groups (P < 0.01). Reactive hypertension occurred in 9 patients (45%) in the 30 mg group (mean highest SAP 120% of baseline, range 104%-143%) compared with 2 (10%), 1 (5%), and 5 (25%) patients in the other groups (P = 0.009). Heart rate changes, total ephedrine requirement, incidence of nausea and vomiting, and neonatal outcome were similar among groups. The proportion of patients with umbilical arterial pH < 7.2 was 10.5%, 25%, 42%, and 22% in the control, 10-, 20 , and 30-mg groups, respectively (P = 0. 12). We conclude that the smallest effective dose of ephedrine to reduce the incidence of hypotension was 30 mg. However, this dose did not completely eliminate hypotension, nausea and vomiting, and fetal acidosis, and it caused reactive hypertension in some patients. IMPLICATIONS: We investigated different doses of IV ephedrine as prophylaxis for hypotension during spinal anesthesia for cesarean delivery and found that the smallest effective dose was 30 mg. However, this dose did not completely eliminate hypotension, caused reactive hypertension in some patients, and did not improve neonatal outcome. PMID- 10825327 TI - Lumbar sympathetic block for sympathetically maintained pain: changes in cutaneous temperatures and pain perception. AB - Lumbar sympathetic block (LSB) is used in the management of sympathetically maintained pain states. We characterized cutaneous temperature changes over the lower extremities after LSB. Additionally, we examined the effects of iohexol, a radio-opaque contrast medium, on temperature changes and pain relief. After institutional review board approval and written, informed consent, 28 LSBs were studied in 17 patients. Iohexol or normal saline was injected in a randomized, double-blinded fashion before bupivacaine. Lower extremity cutaneous temperatures were measured. Pain, allodynia, interference with daily function, and perceived pain relief were reported in a subset of 15 LSBs for 1 wk after the block. The distal lower extremity ipsilateral to the LSB had the greatest magnitude (8.7 degrees +/- 0.8 degrees C) and rate (1.1 degrees +/- 0.2 degrees C/min) of temperature change. The great toe temperature was within 3 degrees C of core temperature within 35 min after LSB. There were no differences in temperature change between the groups. The iohexol group had greater relief of pain until the morning of the first postblock day (P = 0.002) and longer perceived relief of pain (P = 0.01). The maximum temperature of the great toe correlated with allodynia relief (P = 0.0007). Thus clinicians should expect ipsilateral toe temperatures to increase to within approximately 3 degrees C of core temperature. Iohexol does not alter the efficacy of LSB and may improve relief of symptoms. The magnitude of temperature change may predict relief of allodynia. IMPLICATIONS: Cutaneous toe temperatures approaching core temperature provide a useful monitor of lumbar sympathetic block and may predict relief of sympathetically maintained pain. Iohexol will not compromise temperature changes or pain relief. PMID- 10825328 TI - Thoracic paravertebral block for breast surgery. AB - Cosmetic and reconstructive breast augmentation is a frequently performed surgical procedure. Despite advances in medical treatment, surgical intervention is often associated with postoperative pain, nausea, and vomiting. Paravertebral nerve block (PVB) has the potential to offer long-lasting pain relief and fewer postoperative side effects when used for breast surgery. We compared thoracic PVB with general anesthesia for cosmetic breast surgery in a single-blinded, prospective, randomized study of 60 women scheduled for unilateral or bilateral breast augmentation or reconstruction. Patients were assigned (n = 30 per group) to receive a standardized general anesthetic (GA) or thoracic PVB (levels T1-7). Procedural data were collected, as well as verbal and visual analog pain and nausea scores. Verbal postoperative pain scores were significantly lower in the PVB group at 30 min (P = 0.0005), 1 h (P = 0.0001), and 24 h (P = 0.04) when compared with GA. Nausea was less severe in the PVB group at 24 h (P = 0.04), but not at 30 min or 1 h. We conclude that PVB is an alternative technique for cosmetic breast surgery that may offer superior pain relief and decreased nausea to GA alone. IMPLICATIONS: Paravertebral nerve block has the potential to offer long-lasting pain relief and few postoperative side effects when used for breast surgery. We demonstrated that paravertebral nerve block, when compared with general anesthesia, is an alternative technique for breast surgery that may offer pain relief superior to general anesthesia alone. PMID- 10825329 TI - The assessment of dermatomal level of surgical anesthesia after spinal tetracaine. AB - Transcutaneous electrical stimulation (TES), a 60-mA, 50-Hz continuous square wave, has been considered equivalent to surgical incision. We examined whether TES at a smaller current (10 mA) can be used to predict surgical anesthesia and compare the results with sensory block to cold, pinprick, and touch after the administration of spinal tetracaine. Two groups of 40 consecutive patients, 17-69 yr old and 70 yr old or older received a subarachnoid injection of 0. 5% tetracaine in 10% glucose or saline according to the type of surgery. Patients undergoing abdominal surgery received glucose solution, and those scheduled for lower extremities surgery received saline solution, and thus, the resultant four groups of patients were studied. Neural block was assessed by the loss of sensation to cold, pinprick, touch, and TES at 10 mA (T10s), and tolerance (i.e., the loss of pain or discomfort) to TES at 10 (T10p) and 60 (T60) mA. Dermatomal levels of sensory block to cold, pinprick, and touch that were cephalad to T60 varied widely. In contrast, dermatomal levels of T10s and T10p cephalad to T60 were less variable, and the difference between T10s and T60 was the smallest among all the differences in any groups. Our results demonstrate that, regardless of patient age and baricity of a local anesthetic solution, T10s is a good predictor of T60 equivalent to the dermatomal level of surgical anesthesia. IMPLICATIONS: Our results show that the loss of sensation to transcutaneous electrical stimulation at 10 mA, but not cold, pinprick, or touch, is a good predictor of the dermatomal level of block to transcutaneous electrical stimulation at 60 mA, which is considered equivalent to the dermatomal level of surgical anesthesia after the administration of spinal anesthesia. PMID- 10825330 TI - The effects of tramadol and morphine on immune responses and pain after surgery in cancer patients. AB - There has been growing interest in determining the possible immune consequences of opioid administration for the management of postoperative pain. We studied the effects of morphine and tramadol on pain and immune function during the postoperative period in 30 patients undergoing abdominal surgery for uterine carcinoma. Phytohemoagglutinin-induced T lymphocyte proliferation and natural killer cell activity were evaluated immediately before and after surgery, and 2 h after the acute administration of either 10 mg of morphine IM or 100 mg tramadol IM for pain. In all patients, phytohemagglutinin-induced lymphoproliferation was significantly depressed by surgical stress. However, in the morphine-treated group, proliferative values remained lower than basal levels for 2 h after treatment, whereas in tramadol-administered patients proliferative values returned to basal levels. Natural killer cell activity was not significantly affected by surgery nor by morphine administration, whereas tramadol significantly enhanced the activity of natural killer cells. Both drugs produced a comparable reduction in postoperative pain. We conclude that, as previously observed in the experimental animal, tramadol and morphine, when administered in analgesic doses, induce different immune effects. IMPLICATIONS: Recent studies suggest that opioids can have an adverse impact on the immune system. Because surgical stress also induces immune dysfunction, the search for analgesic drugs devoid of immunosuppressive effects is of import. This study compared the effects on immune responses of morphine and of the atypical opioid analgesic, tramadol, given for postoperative pain to gynecological cancer patients. Tramadol and morphine showed comparable analgesic activity; however, tramadol, in contrast to morphine, induced an improvement of postoperative immunosuppression and, therefore, may be preferred to morphine for the treatment of postoperative pain. PMID- 10825331 TI - Acute respiratory insufficiency during interscalene plexus block. PMID- 10825332 TI - Acute vasospastic attack after extradural block in a patient with Raynaud's disease. AB - IMPLICATIONS: Raynaud's disease is an episodic and debilitating disease of the circulation to the fingers and toes. This case report describes an acute exacerbation of Raynaud's disease in a patient undergoing surgery with epidural anesthesia. This potentially hazardous complication has not been previously reported. PMID- 10825333 TI - Does ketamine have preemptive effects in women undergoing abdominal hysterectomy procedures? AB - Ketamine may produce "preemptive" analgesia when administered before surgically induced trauma. Therefore, we hypothesized that pre- versus postincisional administration of ketamine would improve pain control after abdominal hysterectomy procedures. Eighty-nine patients were randomly assigned to one of three treatment groups according to a placebo-controlled, double-blinded protocol: Group 1 (placebo) received saline 0.04 mL/kg IV immediately before and after surgery; Group 2 (preincision), received ketamine 0.4 mg/kg IV before skin incision and saline at the end of the operation; and Group 3 (postincision), received saline before skin incision, and ketamine 0.4 mg/kg IV was given after skin closure. The general anesthetic technique was standardized in all three treatment groups. During the first postoperative hour, Group 3 experienced significantly less pain than Groups 1 and 2, as assessed by using both visual analog and verbal rating scales. There were no significant differences between Groups 1 and 2 with respect to pain scores, postoperative opioid analgesic requirements, and incidence of postoperative nausea and vomiting. We conclude that a single dose of ketamine 0.4 mg/kg IV fails to produce preemptive analgesic effects. IMPLICATIONS: Even though ketamine 0.4 mg/kg IV has short-lasting acute analgesic effects, it failed to produce a preemptive effect when given before abdominal hysterectomy procedures. PMID- 10825334 TI - Ondansetron given before induction of anesthesia reduces shivering after general anesthesia. AB - The neurotransmitter pathways involved in the mechanism of postanesthetic shivering (PAS) are poorly understood. Meperidine, clonidine, and physostigmine are all effective treatments, indicating that opioid, alpha(2)-adrenergic, and anticholinergic systems are probably involved. We investigated the effect of ondansetron, a 5-HT(3) antagonist used to treat postoperative nausea and vomiting, on intraoperative core and peripheral temperatures and PAS. Eighty-two patients (age, 18-60 yr) undergoing orthopedic, general, or urological surgery were randomized into three groups in this double-blinded, placebo-controlled, study: Group O4 (n = 27) received ondansetron 4 mg IV, Group O8 (n = 27) received ondansetron 8 mg IV, and Group C (n = 28) received saline IV immediately before the anesthetic induction. Core (tympanic) and fingertip temperature (dorsum of middle finger) were recorded. Anesthesia was induced with IV fentanyl 1 microg/kg and propofol 2.0-2.5 mg/kg and maintained with 1 minimum alveolar anesthetic concentration isoflurane in 70% nitrous oxide/oxygen. The occurrence of shivering was documented clinically during recovery by nursing staff, who were unaware of the group assignment. PAS occurred in 16 of 28 (57%) patients in Group C, compared with 9 of 27 (33%) in Group O4 (P = 0.13) and 4 of 27 (15%) patients in Group O8 (P = 0.003). Within each group, core temperature decreased and peripheral temperature increased significantly, but there were no significant differences among the groups at any time interval. We conclude that ondansetron 8 mg IV given during the induction of anesthesia prevents PAS without affecting the core-to-peripheral redistribution of heat during general anesthesia. This suggests that serotonergic pathways have a role in the regulation of PAS. IMPLICATIONS: In a randomized, double-blinded, placebo-controlled, clinical study, ondansetron 8 mg IV, given just before the induction, reduced the incidence of postanesthetic shivering compared with saline. The anticipated core to-peripheral redistribution of body temperature during general anesthesia was not affected. This implies that ondansetron probably acts by a central inhibitory mechanism, and that 5-hydroxytryptaminergic pathways have a role in regulating postanesthetic shivering. PMID- 10825335 TI - Absorbents differ enormously in their capacity to produce compound A and carbon monoxide. AB - Concern persists regarding the production of carbon monoxide (CO) and Compound A from the action of carbon dioxide (CO(2)) absorbents on desflurane and sevoflurane, respectively. We tested the capacity of eight different absorbents with various base compositions to produce CO and Compound A. We delivered desflurane through desiccated absorbents, and sevoflurane through desiccated and moist absorbents, then measured the resulting concentrations of CO from the former and Compound A from the latter. We also tested the CO(2) absorbing capacity of each absorbent by using a model anesthetic system. We found that the presence of potassium hydroxide (KOH) and sodium hydroxide (NaOH) increased the production of CO from calcium hydroxide (Ca[OH](2)) but did not consistently affect production of Compound A. However, the effect of KOH versus NaOH was not consistent in its impact on CO production. Furthermore, the effect of KOH versus NaOH versus Ca(OH)(2) was inconsistent in its impact on Compound A production. Two absorbents (Amsorb) [Armstrong Medica, Ltd, Coleraine, Northern Ireland], composed of Ca(OH)(2) plus 0.7% polyvinylpyrrolidine, calcium chloride, and calcium sulfate; and lithium hydroxide) produced dramatically lower concentrations of both CO and Compound A. Both produced minimal to no CO and only small concentrations of Compound A. The presence of polyvinylpyrrolidine, calcium chloride, and calcium sulfate in Amsorb appears to have suppressed the production of toxic products. All absorbents had an adequate CO(2) absorbing capacity greatest with lithium hydroxide. IMPLICATIONS: Production of the toxic substances, carbon monoxide and Compound A, from anesthetic degradation by carbon dioxide absorbents, might be minimized by the use of one of two specific absorbents, Amsorb (Armstrong Medica, Ltd., Coleraine, Northern Ireland) (calcium hydroxide which also includes 0.7% polyvinylpyrrolidine, calcium chloride, and calcium sulfate) or lithium hydroxide. PMID- 10825336 TI - High frequency jet ventilation in interventional fiberoptic bronchoscopy. AB - High frequency jet ventilation (HFJV) is a well accepted method for securing ventilation in rigid and interventional bronchoscopy. We describe a technique of HFJV using a 14F nylon insufflation catheter placed in the trachea to support stent implantation or endobronchial balloon dilation in endobronchial stenoses with the flexible fiberscope. One hundred sixty-one cases were treated with either a metal wire stent (n = 105) or with balloon dilation (n = 56). In addition to HFJV, IV anesthesia was applied in 132 cases. Driving pressure was 1125-1275 mm Hg, frequency 80-100/min, and inspiratory:expiratory ratio of 1:2. Fraction of inspired oxygen ranged from 0.3-1.0. The effects on alveolar ventilation were assessed by using blood-gas analysis and continuous monitoring of transcutaneous oxygen and carbon dioxide tension (P(tc)CO(2)). Complications consisted of hypertension (n = 8), hypotension (n = 6), bronchospasm (n = 5), and hypoxia (n = 6). In 52% of the cases, mild hypercarbia (P(tc)CO2 50-60mm Hg) was observed. In two cases, a P(tc)CO(2) > 80 mm Hg resolved spontaneously when the patients returned to normal breathing after intermittent superimposed ventilation with a face mask. During placement of stents in the proximal trachea, the jet catheter had to be withdrawn, resulting in displacement of the catheter into the pharynx in one case, which was managed safely with the bronchoscope. In conclusion, HFJV achieves satisfactory operating conditions and provides adequate gas exchange for interventional bronchoscopic procedures with the fiberscope. IMPLICATIONS: Safe ventilation is desired when performing tracheobronchial stent implantation and balloon dilation with the fiberscope. High frequency jet ventilation, applied with a 14F insufflation catheter through the nasotracheal route, offers safe ventilatory support with minimal complications. This was evaluated in 161 procedures treating benign and malignant airway stenoses. PMID- 10825337 TI - The effect of clonidine pretreatment on the perioperative proinflammatory cytokines, cortisol, and ACTH responses in patients undergoing total abdominal hysterectomy. AB - We investigated the hypothesis that oral clonidine premedication would decrease the release of proinflammatory cytokines interleukin (IL)-6, IL-1beta, and tumor necrosis factor-alpha, and stress hormones cortisol and adrenocorticotropic hormone (ACTH), in patients who underwent total abdominal hysterectomy. Twenty patients were randomly assigned to one of two groups: the control group received no premedication, whereas the treatment group received clonidine 0.15 mg oral dose 90 min before surgery. Venous blood samples were obtained at preinduction time and immediately, 1, and 3 h after the start of surgery. IL-6 increased significantly at 3 h, and cortisol and ACTH at 1 and 3 h after the start of surgery compared with the baseline values within both groups. IL-1beta levels decreased at all times compared with the baseline value in the control group. In the comparison study between the control group and the clonidine group, there was a significant decrease in IL-6 level 3 h after the start of surgery and IL-1beta at preinduction time in the clonidine group, whereas there were no changes in tumor necrosis factor-alpha, cortisol, and ACTH levels. These results show that clonidine modulates the IL-6 response related to surgical stress. IMPLICATIONS: We assessed the effect of the alpha(2)-adrenoreceptor agonist clonidine on the immunoendocrine activity during the perioperative period. Clonidine 0.15 mg given as oral premedication resulted in the reduced Interleukin-6 production in response to total abdominal hysterectomy. PMID- 10825338 TI - Halothane does not decrease amiloride-sensitive alveolar fluid clearance in rabbits. AB - Halothane decreases alveolar fluid clearance (AFC), a function required for efficient gas exchange in the rat. Further, halothane decreases amiloride sensitive Na(+) transport in rat alveolar type II cells, a process responsible for a significant portion of AFC. We tested the hypothesis that halothane would decrease amiloride-sensitive AFC in rabbits. Rabbits anesthetized with 1.8% halothane had 5% albumin in 0.9% NaCl instilled into the right lung with (n = 11) or without (n = 11) 1 mM amiloride present in the instillate. Similarly, animals anesthetized with IV fentanyl and droperidol were administered 5% albumin solution with (n = 11) or without (n = 11) amiloride. At 90 min after instillation, alveolar fluid samples were obtained, and AFC was determined by changes in fluid protein concentration. Rabbits anesthetized with halothane or fentanyl and droperidol in the absence of amiloride had similar AFC values (35% +/- 12% and 35% +/- 7%, respectively, mean +/- SD). Rabbits anesthetized with halothane or fentanyl and droperidol in the presence of amiloride had similar AFC values (20% +/- 10% and 16% +/- 12%, respectively) that were significantly less than the groups not administered amiloride (P < 0.01). Unlike the rat, the ability of the rabbit to clear fluid from the alveolar space through amiloride sensitive pathways is not decreased by halothane anesthesia. IMPLICATIONS: Unlike the rat, the ability of the rabbit to clear fluid from the alveolar space through amiloride-sensitive pathways is not decreased by halothane anesthesia. PMID- 10825339 TI - The stability of remifentanil hydrochloride and propofol mixtures in polypropylene syringes and polyvinylchloride bags at 22 degrees-24 degrees C. AB - IMPLICATIONS: Remifentanil and propofol mixtures may be prepared extemporaneously. This study demonstrated that such mixtures are stable for 0 to 36 h, depending on relative concentrations of the drugs and the storage container used. The most stable mixtures were stored in syringes and used large concentrations of the two drugs. PMID- 10825340 TI - A simple method for detecting plasma propofol. AB - IMPLICATIONS: We measured plasma propofol levels by head space-gas chromatography analysis using solid-phase microextraction. This method saves time, and does not require elimination of interfering substances in the plasma. The coefficient variation was +/-5% to +/-30%. The detection limit was 10 ng/mL. PMID- 10825341 TI - Excessive use of hetastarch: An iatrogenic cause of bleeding and hypocalcemia? PMID- 10825342 TI - Propofol and analgesia. PMID- 10825343 TI - Jaw thrust maneuver for endotracheal intubation using a fiberoptic stylet. PMID- 10825344 TI - Postoperative pulmonary aspiration of gastric contents in an infant after general anesthesia with laryngeal mask airway (LMA) PMID- 10825345 TI - Bronchial injury: An avoidable complication during bronchial intubation. PMID- 10825346 TI - Guest reviewers list- october 1999 to march 2000 PMID- 10825347 TI - P selectin, pioneer cells and the path to inflammation. PMID- 10825348 TI - Surgery for malformations of cortical development causing epilepsy. AB - Malformations of cortical development (MCD) are responsible for many cases of refractory epilepsy in adults and children. The results of surgical treatment are difficult to assess from the published literature. Judging from the limited number of adequately reported cases, approximately 40% of all cases of MCD treated surgically may be rendered seizure-free over a minimum 2-year follow-up period. This figure is the same for focal cortical dysplasia (FCD), the most common variety of MCD in surgical reports. In comparison with outcome for epilepsy associated with hippocampal sclerosis, this figure is low. Part of the difference may be artificial and related to limited reporting. Much of the difference is likely to relate to the complex underlying biology of MCD. Analysis of epileptogenesis in MCD has been undertaken. Different types of MCD have different sequelae. Some varieties are intrinsically epileptogenic; these include FCD and heterotopia. Although in most cases, the visualized MCD lies within the region of brain responsible for generating seizures (the epileptogenic zone), it may not constitute the entire epileptogenic zone in all cases. For polymicrogyria and schizencephaly in particular, the visualized abnormalities are probably not the most important component of the epileptogenic zone. There is evidence that the epileptogenic zone is spatially distributed and also, in some cases, temporally distributed. These findings may explain poor surgical outcome and the inadequacy of current presurgical evaluative methods. New preoperative techniques offer the opportunity of improved presurgical planning and selection of cases more likely to be rendered seizure-free by current surgical techniques. Of paramount importance is improved reporting. The establishment of a central registry may facilitate this aim. Specific recommendations are made for surgical strategies based on current experience and understanding. PMID- 10825349 TI - Differential adhesion molecule requirements for immune surveillance and inflammatory recruitment. AB - Activated CD4 Th1 lymphocytes can enter the brain in the absence of an inflammatory focus. However, the molecular mediators that regulate this early migration of lymphocytes into the brain have remained unclear. We hypothesized that the entry of these 'pioneer' lymphocytes into the brain is regulated by a set of molecular events that are distinct from those used once inflammation has been established. Using cells fluorescently labelled with the lipophilic dye DiI, myelin basic protein (MBP)-specific CD4 lymphocytes that expressed low or high levels of very late antigen-4 (VLA-4) and non-antigen-specific activated splenocytes homed to mouse brain in similar quantities 2 h after adoptive transfer. However, antigen specificity and VLA-4 expression were required for more robust recruitment by 24 h. Immunocytochemistry revealed endothelial and microenvironmental upregulation of vascular cell adhesion molecule (VCAM), intercellular cell adhesion molecule 1 (ICAM-1), MHC class II and interferon gamma 48 h after transfer of MBP-specific cells. In contrast, expression of meningeal and choroid plexus-associated P selectin was upregulated 2 h after adoptive transfer, but not at 48 h. Monoclonal antibody to P selectin, but not to VLA-4, inhibited early migration of high VLA-4-expressing MBP-specific lymphocytes. These results suggest that early migration occurs independent of the lymphocyte integrin VLA-4 and endothelial VCAM, but does require increased surface expression of endothelial P selectin. PMID- 10825350 TI - Autoimmune disease in first-degree relatives of patients with multiple sclerosis. A UK survey. AB - Previous studies examining an association with other autoimmune diseases have suggested the existence of a generalized autoimmune diathesis in patients with multiple sclerosis. We investigated the prevalence of autoimmune disease in first degree relatives of probands with multiple sclerosis using a case-control method. The results show an excess of autoimmune disease within these families, but no significant association was seen with non-autoimmune diseases. The higher risk in multiplex than simplex families suggests an effect of genetic loading. While the increase in risk applies to each autoimmune disease, autoimmune thyroid disease (and Graves' disease in particular) contributes disproportionately to the excess risk. There was no increase in autoimmune disease within patients with multiple sclerosis themselves when compared with the index controls or population data. We conclude that autoimmune disease is more common in first-degree relatives of patients with multiple sclerosis and hypothesize that common genetic susceptibility factors for autoimmunity co-exist with additional disease specific genetic or environmental factors, which determine clinical phenotype in the individual. PMID- 10825351 TI - Levodopa-responsive dystonia. GTP cyclohydrolase I or parkin mutations? AB - Autosomal dominant DOPA-responsive dystonia (DRD) is usually caused by mutation in the gene encoding guanosine triphosphate-cyclohydrolase I (GTPCH I). We studied 22 families with a phenotype of levodopa-responsive dystonia by sequencing the six coding exons, the 5'-untranslated region and the exon-intron boundaries of the GTPCH I gene. Eleven heterozygous mutations were identified, including five missense mutations, one splice site mutation, two small deletions and two nonsense mutations, in 12 families that included 27 patients and 13 asymptomatic carriers. Six mutations were new and five had already been reported. Four of the mutations caused truncation of the GTPCH I protein. One family carried a base-pair change in the 5'-untranslated region, not detected in controls, that could be responsible for the phenotype. Three of the remaining 10 families had deletions in the parkin gene on chromosome 6, underlining how difficult it is to distinguish, in some cases, between DRD and parkin mutations. No mutations were identified in seven families. The clinical spectrum extended from the classical DRD phenotype to parkinsonism with levodopa-induced dyskinesias, and included spastic paraplegia as well as the absence of dystonia. PMID- 10825352 TI - Impaired social response reversal. A case of 'acquired sociopathy'. AB - In this study, we report a patient (J.S.) who, following trauma to the right frontal region, including the orbitofrontal cortex, presented with 'acquired sociopathy'. His behaviour was notably aberrant and marked by high levels of aggression and a callous disregard for others. A series of experimental investigations were conducted to address the cognitive dysfunction that might underpin his profoundly aberrant behaviour. His performance was contrasted with that of a second patient (C.L.A.), who also presented with a grave dysexecutive syndrome but no socially aberrant behaviour, and five inmates of Wormwood Scrubs prison with developmental psychopathy. While J.S. showed no reversal learning impairment, he presented with severe difficulty in emotional expression recognition, autonomic responding and social cognition. Unlike the comparison populations, J.S. showed impairment in: the recognition of, and autonomic responding to, angry and disgusted expressions; attributing the emotions of fear, anger and embarrassment to story protagonists; and the identification of violations of social behaviour. The findings are discussed with reference to models regarding the role of the orbitofrontal cortex in the control of aggression. It is suggested that J.S.'s impairment is due to a reduced ability to generate expectations of others' negative emotional reactions, in particular anger. In healthy individuals, these representations act to suppress behaviour that is inappropriate in specific social contexts. Moreover, it is proposed that the orbitofrontal cortex may be implicated specifically either in the generation of these expectations or the use of these expectations to suppress inappropriate behaviour. PMID- 10825353 TI - The impact of deep brain stimulation on executive function in Parkinson's disease. AB - Deep brain stimulation (DBS) of the subthalamic nucleus (STN) or the internal segment of the globus pallidus (GPi) improves Parkinson's disease and increases frontal blood flow. We assessed the effects of bilateral DBS on executive function in Parkinson's disease patients, seven with electrodes implanted in the STN and six in the GPi. Patients were assessed off medication with stimulators off, on and off again. The groups showed differential change with stimulation on the Reitan Trail-Making test (TMT B) (STN more improved) and on some measures of random number generation and Wisconsin Card Sorting (STN improved, GPi worse with stimulation). Across the groups, stimulation speeded up responding (Stroop control trial, TMT A) and improved performance on paced serial addition and missing digit tests. Conversely, conditional associative learning became more errorful with stimulation across the two groups. In general, change in performance with stimulation was significant for the STN but not the GPi group. These results support two opposite predictions. In support of current models of Parkinson's disease, 'releasing the brake' on frontal function with DBS improved aspects of executive function. Conversely, disruption of basal ganglia outflow during DBS impaired performance on tests requiring changing behaviour in novel contexts as predicted by Marsden and Obeso in 1994. PMID- 10825354 TI - Reduced striatal dopamine transporters in idiopathic rapid eye movement sleep behaviour disorder. Comparison with Parkinson's disease and controls. AB - Rapid eye movement (REM) sleep behaviour disorder (RBD) is characterized by complex behaviour during REM sleep. The aetiology of this disorder is still unknown, but a recent study showed an association between RBD and Parkinson's disease. We therefore studied striatal postsynaptic dopamine D2 receptor density with [123I](S)-2-hydroxy-3-iodo-6-methoxy-(1-ethyl-2-pyrrolidinylmethyl ) benzamide ([123I]IBZM) and the striatal presynaptic dopamine transporter with (N) (3-iodopropene-2-yl)-2beta-carbomethoxy-3beta-(4-chlorop henyl) tropane ([123I]IPT) using single-photon emission computed tomography (SPECT) in patients with idiopathic RBD. We compared the [123I]IPT-SPECT results of five patients with polysomnographically confirmed idiopathic RBD with the [123I]IPT-SPECTs of seven age- and sex-matched controls without a history of sleep disorders, and of 14 patients with Parkinson's disease (Hoehn and Yahr stage I). All RBD patients had significantly reduced striatal [123I]IPT binding compared with the controls (RBD: right, 2.94 +/- 0.32, left, 3.03 +/- 0.41; controls: right, 4.41 +/- 0.17, left, 4.34 +/- 0.21; P = 0.003), but significantly higher striatal [123I]IPT binding compared with the striatum contralateral to the symptomatic body side of the Parkinson's disease patients (Parkinson's disease: ipsilateral, 3.17 +/- 0.36, P = 0.298; contralateral, 2.51 +/- 0.31, P = 0.019). Uptake of [123I]IBZM was not significantly different in the RBD group compared with the controls. This study demonstrates that [123I]IPT-SPECT is a useful diagnostic tool in RBD and that reduced striatal dopamine transporters may be a pathophysiological mechanism of idiopathic RBD. (Results are given as mean +/- standard deviation.) PMID- 10825355 TI - Human corticospinal excitability evaluated with transcranial magnetic stimulation during different reaction time paradigms. AB - The aim of this study was to evaluate corticospinal excitability of both hemispheres during the reaction time (RT) using transcranial magnetic stimulation (TMS). Nine right-handed subjects performed right and left thumb extensions in simple (SRT), choice (CRT) and go/no-go auditory RT paradigms. TMS, inducing motor-evoked potentials (MEPs) simultaneously in the extensor pollicis brevis muscles bilaterally, was applied at different latencies from the tone. For all paradigms, MEP amplitudes on the side of movement increased progressively in the 80-120 ms before EMG onset, while the resting side showed inhibition. The inhibition was significantly more pronounced for right than for left thumb movements. For the left SRT, significant facilitation occurred on the right after EMG onset. Initial bilateral facilitation occurred in SRT trials with slow RT. After no-go tones, bilateral inhibition occurred at a time corresponding to the mean RT to go tones. The timing of the corticospinal rise in excitability on the side of movement was independent of task difficulty and RT. This suggests that corticospinal activation is, to some extent, in series and not in parallel with stimulus processing and response selection. Corticospinal inhibition on the side not to be moved implies that suppression of movement is an active process. This inhibition is more efficient for right- than for left-side movements in right handed subjects, possibly because of left hemispheric dominance for movement. PMID- 10825356 TI - Acute axonal injury in multiple sclerosis. Correlation with demyelination and inflammation. AB - Damage to axons is taken as a key factor of disability in multiple sclerosis, but its pathogenesis is largely unknown. Axonal injury is believed to occur as a consequence of demyelination and was recently shown to be a feature even of the early disease stages. The present study was aimed at characterizing the association of axonal injury and histopathological hallmarks of multiple sclerosis such as demyelination, cellular infiltration and expression of inflammatory mediators. Therefore, axon reduction and signs of acute axonal damage were quantified in early lesion development of chronic multiple sclerosis and correlated with demyelinating activity and inflammation. Patients with secondary progressive multiple sclerosis revealed the most pronounced axonal injury, whereas primary progressive multiple sclerosis patients surprisingly showed relatively little acute axonal injury. Acute axonal damage, as defined by the accumulation of amyloid precursor protein (APP), was found to occur not only in active demyelinating but also in remyelinating and inactive demyelinated lesions with a large inter-individual variability. Only few remyelinating lesions were adjacent to areas of active demyelination. In this minority of lesions, axonal damage may have originated from the neighbourhood. APP expression in damaged axons correlated with the number of macrophages and CD8-positive T lymphocytes within the lesions, but not with the expression of tumour necrosis factor-alpha (TNF-alpha) or inducible nitric oxide synthase (iNOS). Axonal injury is therefore, at least in part, independent of demyelinating activity, and its pathogenesis may be different from demyelination. This has major implications for therapeutic strategies, which aim at preventing both demyelination and axonal loss. PMID- 10825357 TI - Single word reading in developmental stutterers and fluent speakers. AB - Ten fluent speakers and nine developmental stutterers read isolated nouns aloud in a delayed reading paradigm. Cortical activation sequences were mapped with a whole-head magnetoencephalography system. The stutterers were mostly fluent in this task. Although the overt performance was essentially identical in the two groups, the cortical activation patterns showed clear differences, both in the evoked responses, time-locked to word presentation and mouth movement onset, and in task-related suppression of 20-Hz oscillations. Within the first 400 ms after seeing the word, processing in fluent speakers advanced from the left inferior frontal cortex (articulatory programming) to the left lateral central sulcus and dorsal premotor cortex (motor preparation). This sequence was reversed in the stutterers, who showed an early left motor cortex activation followed by a delayed left inferior frontal signal. Stutterers thus appeared to initiate motor programmes before preparation of the articulatory code. During speech production, the right motor/premotor cortex generated consistent evoked activation in fluent speakers but was silent in stutterers. On the other hand, suppression of motor cortical 20-Hz rhythm, reflecting task-related neuronal processing, occurred bilaterally in both groups. Moreover, the suppression was right-hemisphere dominant in stutterers, as opposed to left-hemisphere dominant in fluent speakers. Accordingly, the right frontal cortex of stutterers was highly active during speech production but did not generate synchronous time-locked responses. The speech-related 20-Hz suppression concentrated in the mouth area in fluent speakers, but was evident in both the hand and mouth areas in stutterers. These findings may reflect imprecise functional connectivity within the right frontal cortex and incomplete segregation between the adjacent hand and mouth motor representations in stutterers during speech production. A network including the left inferior frontal cortex and the right motor/premotor cortex, likely to be relevant in merging linguistic and affective prosody with articulation during fluent speech, thus appears to be partly dysfunctional in developmental stutterers. PMID- 10825358 TI - Movement-related change of electrocorticographic activity in human supplementary motor area proper. AB - We investigated movement-related change in the cortical EEG signal by simultaneous recording from the primary sensorimotor area (S1-M1) and the supplementary motor area proper (SMA proper) in four patients with intractable partial epilepsy. By the use of temporal spectral evolution (TSE) analysis, the change in background cortical activity in relation to self-paced finger/wrist extension was compared among the SMA proper, S1 and M1. All three areas showed a decrease in the amount of activity for the frequency range between 10 and 40 Hz before the onset of movement [event-related desynchronization (ERD)]. The SMA proper showed earlier onset of ERD for 18-22 Hz activity (-3.4 +/- 0.5 s, mean +/ standard deviation) than M1 (-1.7 +/- 0.7 s) and S1 (-1.4 +/- 0.5 s). The degree of ERD in S1 was greatest for 10-14 Hz and that in M1 for 18-22 Hz, whereas in the SMA proper ERD was observed throughout the frequency bands from 10 to 40 Hz. Neither the degree nor the onset time of ERD in the SMA proper was lateralized to either the ipsilateral or the contralateral side with respect to the movement. A transient increase in activity after movement [event-related synchronization (ERS)] was observed in all three areas. In the SMA proper, two out of four subjects showed ERS for frequency bands below 40 Hz with both ipsilateral and contralateral movements. By contrast, in S1 and M1, ERS was recorded for frequency bands between 20 and 90 Hz, and was predominantly associated with the contralateral movement. The present study suggests that the background cortical activity in the SMA proper has a specific temporal pattern with respect to self paced movement, and that the SMA proper is involved in motor preparation earlier than S1-M1 in a bilaterally organized manner. PMID- 10825359 TI - Self-initiated versus externally triggered movements. II. The effect of movement predictability on regional cerebral blood flow. AB - Event-related potential studies in man suggest a role for the supplementary motor area (SMA) in movement preparation, particularly when movements are internally generated. In a previous study combining PET with recording of movement-related cortical potentials, we found similar SMA activation and early pre-movement negativity during self-initiated and predictably paced index finger extensions. Early pre-movement negativity was absent when finger movements were paced by unpredictable cues. We postulated that preparation preceding self-initiated and predictably cued movements was responsible for equivalent levels of SMA activation in these two conditions. To test this, we have performed further studies on six normal volunteers with H2(15)O-PET. Twelve measurements of regional cerebral blood flow were made in each subject under three conditions: rest; self-initiated right index finger extension at a variable rate of once every 2-7 s; and finger extension triggered by pacing tones at unpredictable intervals (at a rate yoked to the self-initiated movements). Activation associated with these conditions was compared using analysis of covariance and t statistics. Compared with rest, unpredictably cued movements activated the contralateral primary sensorimotor cortex, caudal SMA and contralateral putamen. Self-initiated movements additionally activated rostral SMA, adjacent anterior cingulate cortex and bilateral dorsolateral prefrontal cortex (DLPFC). Direct comparison of the two motor tasks confirmed significantly greater activation of these areas and of caudal SMA in the self-initiated condition. These results, combined with our previous data, suggest that rostral SMA plays a primary role in movement preparation while caudal SMA is a motor executive area. In this experiment and in our earlier study, DLPFC was activated only during the self initiated task, in which decisions were required about the timing of movements. PMID- 10825361 TI - ATP in human skin elicits a dose-related pain response which is potentiated under conditions of hyperalgesia. AB - Despite the considerable interest in the possibility that ATP may function as a peripheral pain mediator, there has been little quantitative study of the pain producing effects of ATP in humans. Here we have used iontophoresis to deliver ATP to the forearm skin of volunteers who rated the magnitude of the evoked pain on a visual analogue scale. ATP consistently produced a modest burning pain, which began within 20 s of starting iontophoresis and was maintained for several minutes. Persistent iontophoresis of ATP led to desensitization within 12 min but recovery from this was almost complete 1 h later. Different doses of ATP were delivered using different iontophoretic driving currents. Iontophoresis of ATP produced a higher pain rating than saline, indicating that the pain was specifically caused by ATP. The average pain rating for ATP, but not saline, increased with increasing current. Using an 0.8 mA current, subjects reported pain averaging 27.7 +/- 2.8 (maximum possible = 100). Iontophoresis of ATP caused an increase in blood flow, as assessed using a laser Doppler flow meter. The increase in blood flow was significantly greater using ATP than saline in both the iontophoresed skin (P < 0.01) and in the surrounding skin, 3 mm outside the iontophoresed area (P < 0.05). The pain produced by ATP was dependent on capsaicin-sensitive sensory neurons, since in skin treated repeatedly with topical capsaicin pain was reduced to less than 25% of that elicited on normal skin (2.1 +/- 0.4 compared with 9.3 +/- 1.5 on normal skin). Conversely, the pain producing effects of ATP were greatly potentiated in several models of hyperalgesia. Thus, with acute capsaicin treatment when subjects exhibited touch evoked hyperalgesia but no ongoing pain, there was a threefold increase in the average pain rating during ATP iontophoresis (22.7 +/- 3.1) compared with pre capsaicin treatment (7.8 +/- 2.6). Moreover, ATP iontophoresed into skin 24 h after solar simulated radiation (2 x minimal erythymic dose) resulted in double the pain rating of normal skin, increasing from 15.3 +/- 4.1 to 32.7 +/- 4.1. The pain response to saline was not significantly altered after UV irradiation at any time-point studied. We conclude that ATP produces pain by activating capsaicin sensitive nociceptive afferents when applied to skin. The possibility that ATP activates nociceptors indirectly via its degradation products cannot be ruled out. The effects of ATP are dose-dependent and responses desensitize only slowly. In inflammatory conditions, ATP may be a potent activator of nociceptors and an endogenous mediator of pain. PMID- 10825360 TI - Muscular dystrophy due to dysferlin deficiency in Libyan Jews. Clinical and genetic features. AB - The cluster in Jews of Libyan origin of limb-girdle muscular dystrophy type 2B due to a dysferlin 1624delG mutation is described. The carrier frequency of this mutation is calculated to be approximately 10% in this population, in which the disease prevalence is at least 1 per 1300 adults. Twenty-nine patients from 12 families were all homozygous for the same mutation. However, clinical features were heterogeneous even within the same family: in half of the patients onset was in the distal muscles of the legs, which is similar to Miyoshi myopathy, while in others onset was in the proximal musculature, which is similar to other forms of limb-girdle dystrophies. Age at onset varied from 12 to 28 years (mean 20.3 +/- 5.5 years). One patient was presymptomatic at age 28 years. Progression was slow regardless of age of onset, patients remaining ambulatory until at least 33 years. Five patients described subacute, painful enlarged calves as an early, unusual feature. The variable features in this ethnic cluster contribute to the definition of the clinical spectrum of dysferlinopathies in general. The cause of the observed heterogeneity remains unclear. PMID- 10825362 TI - Dominant partial epilepsies. A clinical, electrophysiological and genetic study of 19 European families. AB - Nineteen families with autosomal dominant partial epilepsy were analysed clinically and electrophysiologically in detail. Seventy-one patients were studied as well as 33 non-epileptic at-risk family members. We subdivided the families into those with autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE) (n = 8), familial temporal lobe epilepsy (n = 7) and autosomal dominant partial epilepsy with variable foci (n = 4). However, the application of this nosology to certain families was difficult in cases of non-specific or conflicting clinical and electrophysiological evidence. This was underscored by the observation by depth electrode recordings in one patient that a so-called ADNFLE may originate in an extrafrontal area. The evolution of familial partial epilepsies, which exhibit great intrafamilial variability, is not always benign. The level of pharmacoresistance may reach 30%, close to that seen in classical cryptogenic partial epilepsies. The familial character of a partial epilepsy may be unrecognized in small families as some affected members may have only EEG abnormalities and are clinically asymptomatic, which reflects incomplete clinical penetrance. In view of the recent discoveries of mutations in the alpha4 nicotinic acetylcholine receptor subunit in a few families with ADNFLE, this genetic study focused on genes encoding nicotinic receptor subunits and a candidate region on chromosome 10q. No mutation was detected in the alpha4 and 012 nicotinic acetylcholine receptor subunits. Positive but not significant lod scores were obtained in four families with markers from the candidate region on chromosome 10q. PMID- 10825363 TI - A systematic study of visual extinction. Between- and within-field deficits of attention in hemispatial neglect. AB - Mechanisms of visual extinction were investigated in four patients with right hemisphere damage using a partial report paradigm. Different shapes (star or triangle) were displayed in one, two or four possible locations so that double simultaneous stimuli occurred either across the two hemifields or within the same hemifield. Patients attended either to the location (right, left or both), number (one, two or four) or shape (no, one or two stars among the shapes presented) of stimuli in three separate experiments using the same displays and exposure duration. Reporting the location (Experiment 1) produced marked contralesional extinction, although reaction time was delayed compared with unilateral right trials, indicating unconscious processing. Reaction time was also delayed on correct bilateral and unilateral left trials. In contrast, enumerating stimuli (Experiment 2) caused no significant contralesional extinction on bilateral displays and reaction time was similar on bilateral and unilateral right trials, suggesting that information from both fields was grouped in a single numerable percept in this task. However, patients often detected only one of two stimuli within the left field. Whereas similarity of shapes improved localization and did not affect enumeration, identifying stars among shapes (Experiment 3) revealed a severe inability to detect two similar targets between hemifields as well as within each of the hemifields. Distracting triangles were generally less detrimental to the perception of a concurrent target on either side, but slowed the reaction time regardless of whether they were in the same or the opposite field. Relative difficulty in ignoring distractors correlated with neglect severity on a cancellation task, and was most prominent in one patient with a large amount of frontal damage. These findings suggest that (i) allocation of attention to identical stimuli can be modulated by task demand; (ii) enumerating a small set of items across fields may not require attending to individual stimuli but relies on preattentive subitizing ability, as found in normal subjects; (iii) location information may be critical for attentional mechanisms subserved by the parietal cortex and pathological competition for awareness in extinction; (iv) extinction entails a bilateral deficit in attending to two concurrent similar targets when their features must be identified; and (v) the relevance of the stimuli can modulate the distribution of attention, possibly through frontal top-down control. These findings are consistent with recent neurophysiological evidence of parietal and frontal attentional influences on ventral visual pathways. PMID- 10825364 TI - Presence of N-cadherin transcripts in mature spermatozoa. AB - The essential mechanism involved in sperm-oolemma fusion has yet to be elucidated. Recognition and binding is initiated by specific cell surface receptor engagement between gametes. Fusion between hamster oolemma and spermatozoa is prevented in the presence of trypsin in Ca(2+)-free media, as is oocyte activation, implicating a cadherin-like adhesion. Cadherins are a family of Ca(2+)-dependent adhesion molecules that bind homotypically with their target, are morphoregulatory and function eptopically to affect tissue form and function. Cadherins and cadherin-associated molecules have been identified in testes and germinal cells, as well as ejaculated spermatozoa. Moreover, cadherins are also present in oocytes and may suggest a cadherin-mediated adhesion in sperm-oocyte interaction. We have detected antigenic epitopes recognized by N-cadherin monoclonal antibodies diffusely distributed over the entire sperm head. In addition, Western blot analysis confirmed the presence of an antibody reactive peptide in spermatozoa, testis and ovary protein extracts at the expected molecular weight for authentic N-cadherin. Total RNA was isolated from mature motile spermatozoa, as well as ovary and testis tissue, and served as template for reverse transcription-polymerase chain reaction (RT-PCR) with N-cadherin specific primers. Alignment of sequences from PCR products of testis, ovary and spermatozoa with published N-cadherin sequence was identical except for occasional base changes. We intend to develop methods to analyse this transcript from small numbers of spermatozoa from a variety of donors to determine if defects in cadherin distribution or structure may predict reduced male fertility. PMID- 10825365 TI - TGFbeta receptor types I and II and the substrate proteins Smad 2 and 3 are present in human oocytes. AB - We have recently found that values of the transforming growth factor (TGF)beta1 in human ovarian follicular fluid obtained during ovarian stimulation for IVF were higher in women who subsequently became pregnant following embryo transfer. We therefore postulated that TGFbeta1 may have a beneficial effect on the preimplantation embryo and improve the chances of a successful implantation. We have used reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry to investigate the presence in human oocytes and preimplantation embryos of the essential components of the TGFbeta signalling pathway, TGFbeta receptors type I and II and the substrate proteins Smad 2 and 3. We found that both receptors, as well as Smad 2 and 3, were present in the unfertilized oocyte, whereas only the type I receptor and Smad 2 and 3 were present at the blastocyst stage. At the 4-cell and 8-cell stages neither of the receptors was present, but Smad 2 and 3 were present at both stages. These findings support our hypothesis that the TGFbeta1 in follicular fluid may interact with the oocyte and preimplantation embryo via TGFbeta receptors, and that TGFbeta signalling may be important for the development of the oocyte and the preimplantation embryo. PMID- 10825366 TI - Spontaneous luteinization of antral marmoset follicles in vitro. AB - Large non-luteinized follicles of the marmoset monkey were cultured for up to 96 h in the presence of substances that are known to induce luteinization, i.e. LH, transforming growth factor (TGF)-beta and cyclic AMP. The state of the basal lamina, and the expression of connexin-43, alpha(2) integrin subunit and TGF-beta receptor type II (TbetaR-II) were chosen as parameters to judge the progress of luteinization. Antral follicles, cultured for 1 h, were not luteinized, as shown by an intact basal lamina, strong immunoreactivity of connexin-43 in granulosa cells, and no expression of TbetaR-II in the theca layer. After 12 h, most follicles showed a dissolution of the basal lamina, a faint reactivity of connexin-43, high expression of TbetaR-II in theca- and outer granulosa cells and high expression of alpha(2) integrin subunit in granulosa cells bordering at the basement membrane; all of which indicate luteinization. After 96 h of culture, luteal structures (e.g. corpora lutea accessoria) had developed. This was true for both non-stimulated and stimulated follicles. Our results strongly suggest that antral follicles luteinize spontaneously. The decisive determinant appears to be the follicular stage. PMID- 10825367 TI - Cytoskeletal organization defects and abortive activation in human oocytes after IVF and ICSI failure. AB - In this study, we analysed the distribution of beta tubulins to detect spindle and cytoplasmic microtubules, alpha acetylated tubulins for sperm microtubules and chromatin configuration in oocytes showing fertilization failure after conventional IVF or intracytoplasmic sperm injection (ICSI). A total of 450 human oocytes that failed to fertilize were studied 20-40 h after IVF or ICSI. In all, 287 oocytes were stained for immunofluorescence and chromosomal spreads were performed by Tarkowski's air-drying method in 163 IVF or ICSI oocytes that did not develop pronuclei after the extrusion of a second polar body. Immunofluorescence analysis showed that the main reason of fertilization failure after IVF was no sperm penetration (55.5%). The remaining oocytes showed different abnormal patterns, e.g. oocyte activation failure (15.1%) and defects in pronuclei apposition (19.2%). On the other hand, fertilization failure after ICSI was mainly associated to incomplete oocyte activation (39.9%), and to a lesser extent with defects in pronuclei apposition (22.6%) and failure of sperm penetration (13.3%). A further 13.3% of the ICSI oocytes arrested their development at the metaphase of the first mitotic division. The chromosomal spreads allowed the analysis of abortive activations, in which no pronuclei formed but a second polar body was extruded. Immunofluorescence and cytogenetic analysis provided a useful tool to improve infertility diagnosis and prognosis in each particular case. PMID- 10825368 TI - Cross-linking of Qa-2 protein, the Ped gene product, increases the cleavage rate of C57BL/6 preimplantation mouse embryos. AB - The Qa-2 protein, a glycosylphosphatidylinositol (GPI)-linked major histocompatibility complex (MHC) Class Ib molecule found on the surface of mouse T-cells and preimplantation embryos, is the product of the preimplantation embryo development (Ped) gene. The Ped gene regulates the rate of early embryonic development and subsequent embryo survival. T-cells treated with anti-Qa-2 monoclonal antibody (mAb) and cross-linked with a secondary antibody, in the presence of a co-stimulatory signal, undergo increased proliferation. The purpose of this study was to determine whether cross-linking of Qa-2 similarly affects preimplantation embryos. We cross-linked Qa-2 protein on the surface of C57BL/6 2 cell and 8-cell embryos, in the presence of 4/5-phorbol-12-myristate-13-acetate (PMA), and assessed the percentage of embryos reaching the blastocyst stage, the percentage hatching from the zona pellucida, [(3)H-thymidine] incorporation into DNA, and the total number of cells per embryo as measures of embryonic cleavage rate. Both 2-cell and 8-cell embryos increased their cleavage rates 48 h after cross-linking of Qa-2, compared with control embryos (P < 0.05). Our results indicate that a Qa-2 protein cross-linking mechanism may be one way by which this protein regulates the rate of preimplantation mouse embryo development. PMID- 10825369 TI - HCG promotes proliferation of uterine leiomyomal cells more strongly than that of myometrial smooth muscle cells in vitro. AB - Uterine myomas often enlarge rapidly during pregnancy. This rapid increase in size may imply that human chorionic gonadotrophin (HCG) influences cell proliferation in uterine leiomyomata. To assess the direct effect of HCG on normal uterine smooth muscle and uterine leiomyomata, we investigated cell proliferation and the expression of cell cycle-related proteins in these cells. Reverse transcription-polymerase chain reaction (RT-PCR) analysis revealed that HCG/LH receptor was present in both cultured myometrial and leiomyomal cells. Treatment with HCG significantly increased cell proliferation in both myometrial and leiomyomal cells (P < 0.03), especially at an early phase in the 9 day culture. The increase in the viable cell number induced by HCG treatment was significantly greater in leiomyoma cells than in myometrial cells on day 3 in culture (P < 0.03). In leiomyomal cells, the expression of proliferating cell nuclear antigen (PCNA), cyclin E and cdc2 was significantly increased by HCG treatment (P < 0.05) even at the lowest concentration used (3 nmol/l). In myometrial cells, the expression of cyclin E and cdc2 was significantly increased by HCG treatment (P < 0.05) only at the highest concentration used (30 nmol/l). These results suggest that HCG directly promotes the proliferation of myometrial and leiomyomal cells, with the latter showing the greater response of the two. PMID- 10825370 TI - Expression of survivin and Bcl-2 in the normal human endometrium. AB - Survivin is a novel inhibitor of apoptosis. It has been reported that survivin is expressed during fetal development and in cancer tissues, but its expression has not been reported in adult tissues. We investigated the expression of survivin in the endometria of women with regular menstrual cycles using reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry, and compared these findings with Bcl-2, an apoptosis inhibitor. Survivin mRNA was detected by RT-PCR in all samples (nine of nine) of endometrium during the secretory phase, but in only four out of seven samples from endometrium during the proliferative phase, and in none of the atrophic endometrium. Immunohistochemistry demonstrated a survivin protein expression that was strongest in the nuclei of glandular epithelial cells during the late secretory phase. In the proliferative phase, glandular epithelial cells were not stained for survivin. The cyclic changes of survivin and Bcl-2 showed an inverse relationship, with Bcl-2 expression being strongest in the proliferative phase and survivin expression being strongest in the secretory phase. The up-regulation of survivin expression may be due to the concurrent rise in progesterone concentrations during the normal menstrual cycle. Moreover, survivin could play an important role independent of Bcl-2 in physiological homeostasis in the normal endometrium. PMID- 10825371 TI - HLA-G expression in extravillous trophoblasts is an intrinsic property of cell differentiation: a lesson learned from ectopic pregnancies. AB - Human leukocyte antigen (HLA)-G is a major histocompatibility gene expressed almost exclusively in extravillous trophoblasts at the fetal-maternal interface. HLA-G may play a role in protecting the fetus from attack by the maternal natural killer cells. The extravillous trophoblasts invade the decidua and maternal spiral arteries. The factors which regulate the cell-specific expression of HLA-G are unknown. In this study we asked if HLA-G is expressed in extravillous trophoblasts that develop outside of their normal cellular environment, as in the case of ectopic pregnancies. Since all ectopic pregnancies implant in the absence of underlying decidua we also used a placenta accreta as an experimental control. We found that HLA-G mRNA and protein were expressed in the extravillous trophoblasts in the 13 ectopic specimens studied. In a case of placenta accreta (which develops without decidua basalis and is therefore adherent to the underlying myometrium), HLA-G mRNA and protein were also expressed. These results suggest that HLA-G expression is induced in a cell autonomous manner rather than determined by appropriate environmental cues. PMID- 10825373 TI - Activation of neurokinin NK(2) receptors by tachykinin peptides causes contraction of uterus in pregnant women near term. AB - The aim of this study was firstly to elucidate whether the mammalian tachykinins substance P (SP), neurokinin A (NKA) and neurokinin B (NKB)-regulated contractility of myometrium obtained from near-term pregnant women, and secondly to investigate the receptor subtype(s) responsible. In the presence of peptidase inhibitors, i.e. thiorphan (3 micromol/l; endopeptidase 24.11 inhibitor), captopril (10 micromol/l; angiotensin converting enzyme inhibitor) and bestatin (10 micromol/l; aminopeptidase inhibitor); all three mammalian tachykinins elicited concentration-related contractions of isolated myometrial preparations. The rank order of agonist potency of the mammalian tachykinins in the presence of the peptidase inhibitors was NKA > SP = NKB, indicating that the contractile effects were mediated by activation of an NK(2) receptor. The NK(2) receptor selective agonist, [Lys(5), MeLeu(9), Nle(10)]NKA(4-10), produced concentration related contractile responses, while the respective NK(1) and NK(3) receptor selective agonists, [Sar(9), Met(O(2))(11)]SP and [N-MePhe(7)]NKB, had no effect either in the absence or presence of the peptidase inhibitors. The NK(2) receptor selective antagonist, SR48968, produced concentration-related rightward shift in the log concentration curve to [Lys(5), MeLeu(9), Nle(10)]NKA(4-10). This study shows that tachykinins elicit contractile effects on human myometrium obtained from pregnant women near term, and that these effects are mediated by an NK(2) receptor. An excitatory effect of the tachykinins on these preparations could indicate a physiological role for these peptides in enhancing contractility of the uterus in women at term. PMID- 10825372 TI - The effect of mifepristone administration on leukocyte populations, matrix metalloproteinases and inflammatory mediators in the first trimester cervix. AB - Cervical ripening is analogous to an inflammatory reaction characterized by an influx of inflammatory cells and an increase in inflammatory mediators. The anti gestogen mifepristone is highly effective in inducing cervical ripening in women throughout gestation. However, its mechanism of action is largely unknown. The aim of the study was to investigate the effect of in-vivo administration of mifepristone on inflammatory cells and mediators in the cervix. Cervical biopsies were taken from women undergoing a first trimester termination of pregnancy at 0, 6, 12, 24 and 36 h (n = 6 per group) after mifepristone administration. Biopsies were fixed for immunohistochemistry and also cultured for subsequent analysis of culture media by radioimmunoassay or enzyme-linked immunosorbent assay. After administration of mifepristone (6-24 h), there was an increase in immunostaining for leukocyte common antigen (CD45), neutrophil elastase, monocytes (CD68), and matrix metalloproteinases (MMP)-1, -8 and -9. Immunostaining for MMP-2 and tissue inhibitor of metalloproteinases (TIMP)-1, -2 and -4 were unaffected by mifepristone treatment. Secretion of monocyte chemotactic protein (MCP-1) was significantly (P < 0.05) increased from biopsies taken 6-24 h after mifepristone administration. Cervical biopsies also released interleukin-8 (IL-8), prostaglandin (PG) E(2), PGF(2alpha) and prostaglandin metabolites (PGEM and PGFM) although their secretion was unaffected by mifepristone treatment. This study suggests that mifepristone may, in part, effect cervical ripening by modulating the influx of inflammatory cells into the cervix, up-regulating MMP expression and inducing chemokine secretion by cervical tissue. PMID- 10825374 TI - beta-endorphin inhibits the production of interleukin-8 by human chorio-decidual cells in culture. AB - Interleukin-8 (IL-8) is produced by human decidual cells in culture, and may play a role in the initiation of parturition. beta-endorphin is released in significant amounts into the maternal and fetal circulation during labour. The effect of beta-endorphin on IL-8 production by human chorio-decidual cells in culture was investigated. Mixed cells were obtained from the decidual surfaces of 35 term placentas. The cells were plated out at 10x10(6) cells per well in Roswell Park Memorial Institute 1640 culture medium. After 48 h the cells were washed and incubated with either plain culture medium (control), 1 micromol/l progesterone, 1-100 nmol/l beta-endorphin or 1 nmol/l N-acetyl beta-endorphin. After 48 h, IL-8 concentrations were measured in the supernatants by enzyme linked immunosorbent assay (ELISA). Experiments were repeated in the presence of naloxone (1 micromol/l) and using calcium-deficient culture medium. Progesterone (P < 0.0002) and beta-endorphin (P < 0. 0005) significantly inhibited the production of IL-8. The inhibitory effect of beta-endorphin was blocked by naloxone and by using calcium-deficient medium. N-acetyl beta-endorphin had no significant effect on IL-8 production. These findings suggest that beta-endorphin has an inhibitory effect on IL-8 production by decidual cells, and that the effect is mediated via opioid receptors and is calcium-dependent. PMID- 10825376 TI - Segregation of sex chromosomes in spermatozoa of 46,XY/47,XXY men by multicolour fluorescence in-situ hybridization. AB - The sex chromosome disomy and diploidy rates on ejaculated spermatozoa from two patients with mosaic Klinefelter's syndrome were estimated, using X/Y/15 multicolour fluorescence in-situ hybridization (FISH). A 8/18 dual fluorescence in-situ hybridization analysis was also carried out. In triple FISH, a total of 1691 (patient 1) and 811 (patient 2) spermatozoa were analysed. Frequencies of cells with hyperhaploidies for sex chromosomes were 2. 01% and 3.45% for patients 1 and 2 respectively, with both patients showing a significantly increased incidence of 24,XY and 24,XX disomies and only patient 2 showing a significantly increased incidence of 24,YY disomy in comparison to the control (P < 0.001). The 46,XX diploidy rate in patient 1 was also significantly higher than the control (P < 0.01). The ratio of X-bearing to Y-bearing spermatozoa differed from the expected 1:1 ratio for only patient 1 (1.18:1). There was no significant difference for chromosomes 8, 15 or 18 disomy frequencies in comparison to those estimated in the control population. These results support the hypothesis that some 47,XXY cells are able to go through meiosis and form spermatozoa with an abnormal gonosomal complement. Thus, there is an increased risk, for these 46,XY/47,XXY men, of producing offspring with a gonosomal abnormality. PMID- 10825375 TI - NF-kappaB and AP-1 are required for cyclo-oxygenase 2 gene expression in amnion epithelial cell line (WISH). AB - Prostaglandins are known to play an important role in human labour and are used clinically to induce labour onset. Cytokines, e.g. interleukin 1 beta (IL-1beta), are up-regulated in the amniotic fluid late in gestation and can increase prostaglandin production through the expression of cyclo-oxygenase 2 (COX-2), the prostaglandin synthetic isoform involved in human labour. We demonstrate in immortalized amnion epithelial (WISH) cells, that IL-1beta causes increased transcription of the COX-2 gene. Luciferase reporter constructs with site directed mutagenesis of the two NF-kappaB sites and an AP-1 site in the COX-2 promoter showed reduced expression of luciferase in transient transfection studies. This suggests that the binding of transcription factors to these sites is essential for the regulation of COX-2 transcription in IL-1beta-treated WISH cells. PMID- 10825377 TI - Prenatal diagnosis of beta-thalassaemia using fetal erythroblasts enriched from maternal blood by a novel gradient. AB - We have assessed a new technique for the isolation of fetal erythroblasts from maternal blood for the non-invasive prenatal diagnosis of pregnancies at risk of beta-thalassaemia. This method relies on the separation of erythroblasts from maternal nucleated cells by a novel step gradient and high speed centrifugation. In four of the six cases examined, single erythroblasts were identified by immunohistochemistry for zeta (zeta) globin. These were individually micromanipulated and analysed by single cell polymerase chain reaction (PCR) and subsequent sequencing of the region of beta-globin locus where the mutations most common to the region of Puglia, Italy, are clustered. In each of the four instances where fetal erythroblasts were identified by antibody staining, the fetal beta-globin genotype was correctly determined. To date, this represents the largest series of non-invasive prenatal diagnoses performed for this haemoglobinopathy. PMID- 10825378 TI - A surface-exposed region of G(salpha) in which substitutions decrease receptor mediated activation and increase receptor affinity. AB - The mechanism by which receptors activate G proteins is unclear because a connection between the receptor and the nucleotide binding site has not been established. To investigate this mechanism, we evaluated the roles in receptor interaction of three potential receptor contact sites in alpha(s): the alpha2/beta4, alpha3/beta5, and alpha4/beta6 loops. Substitutions of alpha(i2) homologs for alpha(s) residues in the alpha2/beta4 loop and alanine substitutions of residues in the alpha4/beta6 loop do not affect activation by the beta(2) adrenergic receptor. However, replacement of five alpha(s) residues in the alpha3/beta5 loop region with the homologous alpha(i2) residues decreases receptor-mediated activation of alpha(s) and increases the affinity of G(s) for this receptor. The substitutions do not alter guanine nucleotide binding or hydrolysis, or activation by aluminum fluoride, indicating that the effects on receptor interaction are not due to a destabilization of the guanine-nucleotide bound state. In a model of the receptor-G protein complex, the alpha3/beta5 loop maps near the second and third intracellular loops of the receptor. The effects of the alpha3/beta5 substitutions suggest that the wild-type residues may be receptor contact sites that are optimized to ensure the reversibility of receptor G protein interactions. Furthermore, the alpha3/beta5 region corresponds to an exchange factor contact site in both EF-Tu and Ras, suggesting that the mechanisms by which seven-transmembrane receptors and exchange factors catalyze nucleotide exchange may share common elements. PMID- 10825379 TI - Dopamine transporter transmembrane domain polar mutants: DeltaG and DeltaDeltaG values implicate regions important for transporter functions. AB - Polar residues in dopamine transporter (DAT) transmembrane domains (TMs) are likely to act individually and even interactively in recognizing cocaine and dopamine. We initially evaluated the effects of alanine substitution mutants that remove the polar side chains from residues in each of the 12 putative DAT TMs on the recognition of dopamine and the cocaine analog CFT. Eleven combination mutants with multiple substitutions in DAT TMs 4, 5, 7, or 11 were then selected as candidates for more detailed evaluation based on mutation effects on dopamine and cocaine analog affinities. An evaluation of Gibbs free energy changes displayed by single and combined TM mutants (DeltaG(o) and DeltaDeltaG(o)(int)) reveals three categories of potential interactions among mutants: 1) independent, noncooperative interactions (five influenced CFT and two influenced dopamine affinities), 2) synergistic influences (two for CFT and four for dopamine), and 3) complementation of influences on CFT recognition (four mutants) or on dopamine affinity (five). Combined mutations in TMs 4 and 5 yield the largest DeltaDeltaG(o)(int) values for dopamine uptake. TMs 4 and 11 mutants provide the largest DeltaDeltaG(o)(int) for CFT binding. Interactions between residues lying in DAT TMs 4 and 5 support current DAT structural models that suggest the juxtaposition of these two TMs. These data also support contributions of TM 4 and 11 residues to a polar pocket important for cocaine recognition. These candidate interactive DAT polar domains provide larger target sites for compounds that could modulate specific DAT functions than those provided by single mutations alone. PMID- 10825380 TI - Late endosomal/lysosomal targeting and lack of recycling of the ligand-occupied endothelin B receptor. AB - A fusion protein consisting of the endothelin B (ET(B)) receptor and the enhanced green fluorescent protein (EGFP) in conjunction with Cyanin3- or fluorescein conjugated endothelin 1 (Cy3-ET1, Fluo-ET1) was used to investigate the ligand mediated internalization of the ET(B) receptor. The ET(B) receptor and the ET(B)/EGFP fusion protein displayed very similar pharmacological properties when expressed in Chinese hamster ovary cells. The integrity of the fusion protein was verified by low temperature PAGE analysis of the (125)I-ET1-bound ET(B) receptor and the (125)I-ET1-bound ET(B)/EGFP fusion protein. Fluorescence microscopy of Chinese hamster ovary cells expressing the ET(B)/EGFP fusion protein demonstrated strong signals at the plasma membrane. On addition of Cy3-ET1, internalization of ligand and receptor occurred within 5 min via a sucrose-sensitive (i.e., clathrin mediated) pathway. On further incubation, ET(B)/EGFP and Cy3-ET1 fluorescences were found in the perinuclear region, colocalized with fluorescent low density lipoproteins, a marker of the late endosomal/lysosomal pathway, but not with fluorescent transferrin, a marker of the recycling pathway. No dissociation of Cy3-ET1 from the receptor was seen within 4 h. Using (125)I-ET1 or Cy3-ET1, binding sites were again demonstrable at the cell surface within 2 h. The reappearance of binding sites was abolished by prior treatment of the cells with cycloheximide, an inhibitor of protein synthesis. The data demonstrate that the ligand-occupied ET(B) receptor is internalized; however, it does not recycle like most of the G protein-coupled receptors but is sorted to the late endosomal/lysosomal pathway in a manner similar to that of the family of protease activated receptors. PMID- 10825381 TI - Probing the role of a conserved M1 proline residue in 5-hydroxytryptamine(3) receptor gating. AB - A conserved proline residue is found in the first transmembrane domain (M1) of every subunit in the ligand-gated ion channel superfamily. The position of this proline between the N-terminal extracellular agonist binding and the second transmembrane (M2) channel lining domains in the primary sequence suggests its possible involvement in the gating of the receptor. Replacing this proline with alanine, glycine, or leucine in the 5-hydroxytryptamine (5-HT)(3A) homomeric receptors expressed in Xenopus laevis oocytes resulted in the absence of 5-HT induced whole-cell currents, although there were normal levels of specific surface [(3)H]granisetron ([(3)H]BRL-43694) binding sites. To determine what properties of the conserved proline are critical for the function of the channel, two imino acids and an alpha-hydroxy acid were incorporated at the proline position using the nonsense suppression method. trans-3-Methyl-proline, pipecolic acid, and leucic acid were able to replace the conserved proline to produce active channels with EC(50) values similar to that for the wild-type receptor. These trends are preserved in the heteromeric receptors consisting of 5-HT(3A) and 5-HT(3B) subunits in oocytes. The prominent common feature among these residues and proline is the lack of hydrogen bond donor activity, potentially resulting in a flexible secondary structure in the M1 region. Thus, lack of hydrogen bond donor activity may be a key element in channel gating and may explain the high degree of conservation of this M1 proline. PMID- 10825382 TI - Post-transcriptional regulation of bradykinin B1 and B2 receptor gene expression in human lung fibroblasts by tumor necrosis factor-alpha: modulation by dexamethasone. AB - The cellular and molecular mechanisms governing bradykinin B1 and B2 receptor expression and function are poorly understood. We investigated the regulation of both B1 and B2 receptors in human embryonic lung fibroblasts (HEL 299) by the proinflammatory cytokines tumor necrosis factor alpha (TNF-alpha) and interleukin 1beta (IL-1beta). TNF-alpha and IL-1beta both induced a rapid and transient increase in B1 and B2 receptor mRNA expression that was maximal by 2 h, accompanied by an increase in B1 and B2 receptor protein, as measured by radioligand binding assay with [(3)H]des-Arg(10)-kallidin, and [(3)H]bradykinin, respectively. The induced B1 receptors were functionally coupled, because the B1 agonist, des-Arg(10)-kallidin, induced an increase in arachidonic acid release in TNF-alpha-stimulated cells but not in control cells. The induction of B1 and the up-regulation of B2 receptors by TNF-alpha was partly mediated through activation of p38 mitogen-activated protein kinase and that of B2 receptor by protein kinase A. TNF-alpha and IL-1beta regulation of both B1 and B2 receptors was inhibited by dexamethasone. When compared with vehicle-treated cells, dexamethasone increased the rate of decline of both B1 and B2 receptor mRNAs. Nuclear run-on experiments demonstrate that the induction of B1 and the up-regulation of B2 receptors as well as the inhibitory effect of dexamethasone are entirely mediated through post transcriptional mechanisms. PMID- 10825383 TI - Reconstitution of the human 5-HT(1D) receptor-G-protein coupling: evidence for constitutive activity and multiple receptor conformations. AB - The 5-hydroxytryptamine (5-HT) 1D/1B receptors have gained particular interest as potential targets for treatment of migraine and depression. G-protein coupling and other intrinsic properties of the human 5-HT(1D) receptor were studied using a baculovirus-based expression system in Sf9 cells. Coexpression of the human 5 HT(1D) receptor with Galpha(i1), alpha(i2), alpha(i3), or Galpha(o)-proteins and Gbeta(1)gamma(2)-subunits reconstituted a Gpp(NH)p-sensitive, high affinity binding of [(3)H]5-HT to this receptor, whereas the Galpha(q)beta(1)gamma(2) heterotrimer was ineffective in this respect. Competition of [(3)H]5-HT binding by various compounds confirmed that coexpression of the human 5-HT(1D) receptor with Galpha(i/o)beta(1)gamma(2) reconstitutes the receptor in a high affinity agonist binding state, having the same pharmacological profile as the receptor expressed in mammalian cells. Binding of the antagonist ocaperidone to the human 5-HT(1D) receptor in coupled or noncoupled state was analyzed. This compound competed with [(3)H]5-HT binding more potently on the human 5-HT(1D) receptor in the noncoupled state, showing its inverse agonistic character. Ocaperidone acted as a competitive inhibitor of [(3)H]5-HT binding when tested with the coupled receptor form but not so when tested with the noncoupled receptor preparation. Finally, [(35)S]GTPgammaS binding experiments using the inverse agonist ocaperidone revealed a high level of constitutive activity of the human 5-HT(1D) receptor. Taken together, the reconstitution of the human 5-HT(1D) receptor-G protein coupling using baculovirus-infected Sf9 cells made possible the assessment of coupling specificity and the detection of different binding states of the receptor induced by G-protein coupling or ligand binding. PMID- 10825384 TI - Coupling of I(1) imidazoline receptors to the cAMP pathway: studies with a highly selective ligand, benazoline. AB - Clonidine and benazoline are two structurally related imidazolines. Whereas clonidine binds both to alpha(2)-adrenoceptors (alpha(2)R) and to I(1) imidazoline receptors (I(1)R), benazoline showed a high selectivity for imidazoline receptors. Although the alpha(2)R are negatively coupled to adenylate cyclase, no effect on cAMP level by activation of I(1)R has been reported so far. We therefore aimed to compare the effects of clonidine and benazoline on forskolin-stimulated cAMP levels in cell lines expressing either I(1)R only (PC12 cells), alpha(2)R only (HT29 cells), or I(1)R and alpha(2)R together (NG10815 cells). Clonidine proved able to decrease the forskolin-stimulated cAMP level in the cells expressing alpha(2)R and this effect could be blocked by rauwolscine. In contrast, in cells lacking these adrenoceptors, clonidine had no effect. On the other hand, benazoline and other I(1) receptor-selective imidazolines decreased forskolin-stimulated cAMP level in the cells expressing I(1)R, in a rauwolscine- and pertussis toxin-insensitive manner. These effects were antagonized by clonidine. According to these results, we demonstrated that 1) alpha(2)R and I(1)R are definitely different entities because they are expressed independently in different cell lines; 2) alpha(2)R and I(1)R are both implicated in the cAMP pathway in cells (one is sensitive to pertussis toxin and the other is not); and 3) I(1)R might be coupled to more then one transduction pathway. These new data will be essential to further understand the physiological implications of the I(1)R and the functional interactions between I(1) receptors and alpha(2)-adrenoceptors. PMID- 10825385 TI - Doxorubicin impairs crossbridge turnover kinetics in skinned cardiac trabeculae after acute and chronic treatment. AB - Crossbridge dynamics underlying the acute and chronic inotropic effects of doxorubicin (Dox) were studied by application of releasing length steps (amplitude, 0.5-10%) to skinned cardiac trabeculae. Acute incubation of trabeculae with 20 microM Dox for 30 min resulted in a decrease of the velocity of unloaded shortening (V(0), from 9.3 +/- 1.1 to 7.7 +/- 0.7 microm/s, P <.05) and in an increase of the rate of force redevelopment (tau(r), from 56 +/- 4 to 65 +/- 3 ms, P <.05) in response to step amplitudes ranging from 5 to 10%. In contrast, chronic Dox treatment in rats (2 mg/kg/week for 4 weeks) significantly impaired trabecular crossbridge dynamics after step releases of 0.5%. This was reflected by an increase of all time constants describing tension recovery: tau(1), from 10 +/- 1 to 14 +/- 1 ms; tau(2), from 65 +/- 6 to 82 +/- 6 ms; tau(3), from 92 +/- 7 to 293 +/- 67 ms; P <.05. In addition, V(0) was decreased (from 8.6 +/- 0.6 to 6.8 +/- 0.3 microm/s, P <.05) and tau(r) was increased (from 67 +/- 4 to 89 +/- 3 ms; P <.05) in the slack-test. We found that chronic Dox treatment resulted in a shift from the "high ATPase" alpha-myosin heavy chain (MHC) isoform toward the "low-ATPase" beta-MHC isoform in the ventricles (control: alpha-MHC 79 +/- 2% and beta-MHC 21 +/- 2%; Dox-treated: alpha-MHC 53 +/- 2% and beta-MHC 47 +/- 2%; P <.05). The present results show that acute Dox incubation affects the detachment rate of crossbridges, which leads to a delayed relaxation and an arrest of crossbridges in strongly bound states. In contrast, chronic Dox treatment leads to an impairment of both the attachment and detachment rates in the crossbridge cycle, which may be explained by an altered MHC isoform composition in ventricular myocardium. Interfering with Dox-induced alterations of crossbridge kinetics may provide a new strategy to prevent Dox associated cardiotoxicity. PMID- 10825386 TI - A repressive cross-regulation between catalytic and promoter activities of the CYP1A1 and CYP2E1 genes: role of H(2)O(2). AB - Cytochrome P450 enzymes catalyze the first step of the metabolism and subsequent elimination of hydrophobic xenobiotics. However, the activity of some isoforms, among them CYP1A1 and CYP2E1, may result in cellular insults such as oxidative stress and activation of procarcinogen compounds into reactive metabolites. The regulation of the expression of these enzymes is therefore important. We have previously shown that the CYP1A1 gene promoter was repressed by oxidative stress. We show here that the CYP2E1 gene promoter is down-regulated by exogenous H(2)O(2) addition and glutathione depletion. It is also repressed by the transfection of a CYP2E1 expression vector, which elicits an intracellular H(2)O(2) generation. This autoregulation is limited by catalase (which catalyzes the catabolism of H(2)O(2)), thus implying H(2)O(2) as a mediator of the negative feedback mechanism. Furthermore, we observed that the activity of CYP1A1 resulting either from the stimulation of the endogenous gene by benzo[a]pyrene treatment or from the transfection of an expression vector, repressed the activity of the CYP2E1 gene promoter. Conversely, CYP2E1 overexpression repressed the activity of the CYP1A1 gene promoter. In both cases, catalase and a specific inhibitor of one enzyme prevented the repression of the other. This suggests that the generation of H(2)O(2) during the catalytic cycle of these enzymes is a mediator of the cross-regulatory mechanisms. These novel repressive mechanisms of autoregulation and cross-regulation using H(2)O(2) as a common mediator may limit the potential toxicity resulting from high cytochrome P450 activity within the cell. PMID- 10825387 TI - Two different signaling mechanisms involved in the excitation of rat sympathetic neurons by uridine nucleotides. AB - UTP stimulates transmitter release and inhibits M-type K(+) channels in rat superior cervical ganglion neurons via G protein-coupled P2Y receptors. To investigate the underlying signaling mechanisms, we treated the neurons with either pertussis or cholera toxin; neither treatment altered the inhibition of M type K(+) channels by 10 microM UTP. However, pertussis toxin reduced UTP-evoked [(3)H]noradrenaline release by 66%. UTP, UDP, ATP, and ADP caused accumulation of inositol trisphosphate in a pertussis toxin-insensitive manner. Pharmacological inhibition of inositol trisphosphate-induced Ca(2+) release (by inhibition of phospholipase C, of inositol trisphosphate receptors, and of the endoplasmic Ca(2+)-ATPase) prevented the UTP-dependent inhibition of M currents but failed to alter UTP-evoked [(3)H]noradrenaline release. Chelation of intracellular Ca(2+) by 1,2-bis(2-aminophenoxy)ethane-N, N,N',N'-tetraacetic acid also reduced the inhibition of M currents by UTP. In addition, all these manipulations attenuated the inhibition of M currents by bradykinin, but hardly affected the inhibitory action of oxotremorine M. These results demonstrate that UTP inhibits M-type K(+) channels via an inositol trisphosphate-dependent signaling cascade that is also used by bradykinin but not by muscarinic acetylcholine receptors. In contrast, the secretagogue action of UTP is largely independent of this signaling cascade but involves pertussis toxin-sensitive G proteins. Thus, UTP-sensitive P2Y receptors excite sympathetic neurons via at least two different signal transduction mechanisms. PMID- 10825388 TI - Cycloheximide increases proenkephalin and tyrosine hydroxylase gene expression in rat adrenal medulla. AB - The effect of cycloheximide (CHX; 5 mg/kg) on proenkephalin (proENK) and tyrosine hydroxylase (TH) mRNA expression in rat central and peripheral nervous systems was studied. CHX increased proENK and TH mRNA levels in the adrenal gland, but not in hippocampus, striatum, midbrain, brainstem, pituitary, and hypothalamus. The pretreatment with actinomycin D (0.5 mg/kg) significantly decreased CHX induced proENK and TH mRNA expression, suggesting that the CHX-dependent increase of these mRNA levels may be caused by the increase of transcriptional activity rather than RNA stabilization. To investigate the factors involved in CHX-induced proENK and TH mRNA expression, the effect of CHX on activator protein-1 (AP-1), cAMP response element (CRE) binding protein (CREB), and glucocorticoid response element (GRE) was tested. In AP-1, the basal expression of Fra-2 and c-Jun proteins and AP-1 DNA binding activity in the adrenal medulla was higher than other tissues tested, but CHX reduced these protein levels and AP-1 DNA binding activity. In CREB, CHX time dependently increased the level of phospho-CREB without altering total CRE level and CRE DNA binding activity. Furthermore, phospho-CREB actively participated in CRE DNA binding activity. In GRE, although CHX increased plasma and adrenal corticosterone level, RU486 (10 mg/kg) reduced CHX-induced proENK, but not TH, mRNA level in a partial manner. These results suggest that the basal expression of proENK and TH mRNA transcription in the adrenal gland seems to be tonically inhibited by de novo protein synthesis. In addition, CHX-dependent increase of proENK and TH mRNA expression in the adrenal medulla is well correlated with phospho-CREB level, but not AP-1. Finally, glucocorticoid seems to be involved at least partially in CHX-dependent proENK, but not TH, mRNA expression in the adrenal medulla. PMID- 10825389 TI - A kinetic binding study to evaluate the pharmacological profile of a specific leukotriene C(4) binding site not coupled to contraction in human lung parenchyma. AB - We report the identification of a novel pharmacological profile for the leukotriene (LT)C(4) binding site we previously identified in human lung parenchyma (HLP). We used a series of classic cysteinyl-LT (CysLT)(1) receptor antagonists belonging to different chemical classes and the dual CysLT(1) CysLT(2) antagonist BAY u9773 for both binding and functional studies. Because the presence of (S)-decyl-glutathione interfered with cysteinyl-LT binding, with a kinetic protocol we avoided the use of this compound. By means of heterologous dissociation time courses, we demonstrated that zafirlukast, iralukast, and BAY u9773 selectively competed only for (3)H-LTD(4) binding sites, whereas pobilukast, pranlukast, and CGP 57698 dissociated both (3)H-LTC(4) and (3)H LTD(4) from their binding sites. Thus, with binding studies, we have been able to identify a pharmacological profile for LTC(4) distinct from that of LTD(4) receptor (CysLT(1)) in HLP. On the contrary, in functional studies, all of the classic antagonists tested were able to revert both LTC(4)- and LTD(4)-induced contractions of isolated HLP strips. Thus, LTD(4) and LTC(4) contract isolated HLP strips through the same CysLT(1) receptor. The results of kinetic binding studies, coupled to a sophisticated data analysis, confirm our hypothesis that HLP membranes contain two cysteinyl-LT high-affinity binding sites with different pharmacological profiles. In functional studies, however, LTD(4)- and LTC(4) induced contractions are mediated by the same CysLT(1) receptor. In conclusion, the specific LTC(4) high-affinity binding site cannot be classified as one of the officially recognized CysLT receptors, and it is not implicated in LTC(4)-induced HLP strip contractions. PMID- 10825390 TI - Identification and molecular characterization of rat CXCR3: receptor expression and interferon-inducible protein-10 binding are increased in focal stroke. AB - We describe here the cloning and characterization of a rat homolog of the chemokine receptor CXCR3. The predicted amino acid sequence of rat CXCR3 contains 367 amino acid residues, sharing 96 and 87% amino acid sequence identity to the murine and human CXCR3, respectively. Among a large panel of chemokines tested, only interferon-inducible protein-10 (IP-10), interferon-gamma-induced monokine, and interferon-inducible T cell alpha-chemoattractant demonstrated specific abilities to induce an intracellular calcium mobilization response in human embryonic kidney 293 cells transfected with rat CXCR3 expression vector. (125)I IP-10 competition binding studies to the CXCR3-transfected human embryonic kidney 293 cells demonstrated that human IP-10 and interferon-inducible T cell alpha chemoattractant are more potent ligands than human interferon-gamma-induced monokine. Following our previous observation for the induced expression of IP-10 in focal stroke, we demonstrate here the time-dependent up-regulation of CXCR3 mRNA in the rat ischemic cortex after permanent occlusion of the middle cerebral artery. A significant increase in (125)I-IP-10-specific binding to ischemic cerebral cortical samples was obtained and paralleled the increase in CXCR3 mRNA expression. The changes in receptor expression and ligand binding correlate highly with known changes in leukocyte accumulation, and gliosis occurred after focal stroke. These data suggest that CXCR3/IP-10 may be a potential novel therapeutic target in focal stroke. In addition, the cloning of rat CXCR3 provides an important tool for the investigation of the pathophysiological role of CXCR3 in other rodent disease models. PMID- 10825391 TI - Glycine and gamma-aminobutyric acid(A) receptor function is enhanced by inhaled drugs of abuse. AB - Inhalable solvents possess significant abuse liability and produce many of the neurobehavioral effects typically associated with central nervous system depressant agents, including motor incoordination, anxiolysis, and the elicitation of signs of physical dependence on withdrawal. We tested the hypothesis that the commonly abused solvents toluene, 1,1,1-trichloroethane (TCE), and trichloroethylene (TCY) affect ligand-gated ion channel activity, as do other classes of central nervous system-depressive agents. TCE and toluene, like ethanol, reversibly enhanced gamma-aminobutyric acid (GABA)(A) receptor mediated synaptic currents in rat hippocampal slices. All three inhalants significantly and reversibly enhanced neurotransmitter-activated currents at alpha1beta1 GABA(A) and alpha1 glycine receptors expressed in Xenopus oocytes. We previously identified specific amino acids of glycine and GABA(A) receptor subunits mediating alcohol and volatile anesthetic enhancement of receptor function. Toluene, TCE, and TCY were tested on several glycine receptor mutants, some of which were insensitive to ethanol and/or enflurane. Toluene and TCY enhancement of glycine receptor function was seen in all these mutants. However, the potentiating effects of TCE were abolished in three mutants and enhanced in two, a pattern more akin to that seen with enflurane than ethanol. These data suggest that inhaled drugs of abuse affect ligand-gated ion channels, and that the molecular sites of action of these compounds may overlap with those of ethanol and the volatile anesthetics. PMID- 10825392 TI - Response element and coactivator-mediated conformational change of the vitamin D(3) receptor permits sensitive interaction with agonists. AB - The vitamin D receptor (VDR) is the nuclear receptor for 1, 25-dihydroxyvitamin D(3) [1alpha,25(OH)(2)D(3)] that acts as a ligand-dependent transcription factor via combined contact with coactivator proteins (steroid receptor coactivator-1, transcriptional intermediary factor 2, and receptor associated coactivator 3) and specific DNA binding sites [vitamin D response elements (VDREs)]. Ligand-mediated conformational changes of the VDR contribute to the key mechanisms in this nuclear hormone signaling process. 1alpha,25(OH)(2)D(3), MC1288 [20-epi 1alpha,25(OH)(2)D(3)], ZK161422 [20-methyl-1alpha,25(OH)(2)D(3)], and Ro27-2310 (also called Gemini, having two side chains at carbon 20) were used as model VDR agonists. The analysis of agonist-induced VDR conformations and coactivator interactions were found to be insufficient for extrapolating in vivo activities. In DNA-independent assays, such as classical limited protease digestions and glutathione S-transferase pull downs, Gemini seemed to be up to 10,000-fold and the other VDR agonists 10- to 100-fold weaker than in functional in vivo assays. A more accurate description of the gene regulatory potential of VDR agonists was obtained with all tested VDR agonists by analyzing VDR conformations in the context of VDRE-bound VDR-retinoid X receptor heterodimers, in such assays as gel supershift, gel shift clipping, and limited protease digestion in the presence of DNA and cofactor. Coactivators were found to shift the ligand sensitivity (by a factor of 4 for Gemini) and the ratio of VDR conformations in the presence of DNA toward the high-affinity ligand binding conformation (c1(LPD)). In conclusion, the induction of response element- and coactivator-modulated VDR conformations appears to be a key step for the gene regulatory function of a VDR agonist. The quantification of these effects would be of central importance for the evaluation of the cell-specific efficacy of systemically applied 1alpha, 25(OH)(2)D(3) analogs. PMID- 10825393 TI - Molecular basis for differential sensitivity of KCNQ and I(Ks) channels to the cognitive enhancer XE991. AB - Channels formed by coassembly of the KCNQ1 (KvLQT1) subunit and the minK subunit underlie slowly activating cardiac delayed rectifier (I(Ks)) in the heart, whereas two other members of the KCNQ channel family, KCNQ2 and KCNQ3, coassemble to underlie the M current in the nervous system. Because of their important physiological function, KCNQ channels have potential as drug targets, and an understanding of possible mechanisms that would enable tissue-specific targeting of these channels will be of significant value to drug development. In this study, we examined the role of the minK subunit in determining the response of KCNQ1 channels to blockade by the cognitive enhancer XE991. Coexpression with minK markedly decreased the sensitivity of KCNQ1 to blockade by XE991. When measured at the end of a 500-ms step, XE991 blockade of the KCNQ1+minK current had a K(D) value of 11.1 +/- 1.8 microM, approximately 14-fold less sensitive than the block of the KCNQ1 current (K(D) = 0.78 +/- 0.05 microM). In addition, XE991 reduced activation and deactivation time constants and caused a rightward shift in the activation curve of KCNQ1+minK, but affected none of these parameters for KCNQ1 alone. Also, XE991 block of KCNQ1+minK, but not of KCNQ1, was time- and voltage-dependent. We conclude that the presence of minK in the I(Ks) channel complex gives rise to differential sensitivity of KCNQ and I(Ks) channels to blockade by XE991. Our results have implications for drug development by demonstrating the important potential role of accessory subunits in determining the pharmacological properties of KCNQ channels. PMID- 10825394 TI - Bryostatin 1 induces prolonged activation of extracellular regulated protein kinases in and apoptosis of LNCaP human prostate cancer cells overexpressing protein kinase calpha. AB - Previously, we reported that 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced apoptosis of LNCaP human prostate cancer cells was accompanied by prolonged translocation of protein kinase C (PKC)alpha to non-nuclear membranes and that TPA-resistant LNCaP cells had down-regulated PKCalpha. Here we show that 10 nM bryostatin 1 induced transient membrane translocation and down-regulation of PKCalpha, prolonged translocation of PKCdelta and epsilon to non-nuclear membranes, and did not induce cell death but blocked TPA-induced apoptosis. To test the hypothesis that inhibition of TPA-induced apoptosis by bryostatin 1 was due to down-regulation of PKCalpha, we inducibly overexpressed PKCalpha in LNCaP cells. Overexpression of PKCalpha alone did not induce apoptosis, even in clones that contained much more membrane-bound, active PKCalpha than was observed in TPA treated untransfected LNCaP cells. However, the addition of 10 nM bryostatin 1 to PKCalpha-overexpressing LNCaP cells did not yield down-regulation of PKCalpha and induced extensive apoptosis. Immunoblot analysis revealed that TPA induced prolonged hyperphosphorylation of Raf-1 and activation of extracellular regulated/mitogen-activated protein kinases 1 and 2 in untransfected LNCaP cells, as did bryostatin 1 in PKCalpha-overexpressing cells. On the other hand, bryostatin 1 induced only transient hyperphosphorylation of Raf-1 and activation of extracellular-regulated/mitogen-activated protein kinases 1 and 2 in untransfected LNCaP cells. These results confirm a role of prolonged membrane associated PKCalpha in PKC activator-mediated LNCaP apoptosis and suggest involvement of the mitogen-activated protein kinase pathway. PMID- 10825395 TI - Identification of a 97-kDa mastoparan-binding protein involving in Ca(2+) release from skeletal muscle sarcoplasmic reticulum. AB - Mastoparan (MP) and radiolabeled [Tyr(3)]MP caused a transient Ca(2+) release from the heavy fraction of sarcoplasmic reticulum, which was inhibited by ryanodine. MP enhanced [(3)H]ryanodine binding in a concentration-dependent manner with an EC(50) value of approximately 0.3 microM. The (45)Ca(2+) release was accelerated by MP, [Tyr(3)]MP, or caffeine in a concentration-dependent manner. The EC(50) values for MP, [Tyr(3)]MP, and caffeine were approximately 2. 0 microM, 7.7 microM, and 1.8 mM, respectively. MP, like caffeine, shifted the stimulatory limb of a bell-shaped curve of Ca(2+) dependence to the left. (45)Ca(2+) release induced by caffeine was completely inhibited by typical blockers of Ca(2+)-induced Ca(2+) release, such as Mg(2+), ruthenium red, or procaine. However, (45)Ca(2+) release induced by MP was completely inhibited by Mg(2+), but it was only partially inhibited by ruthenium red or procaine. The rate of (45)Ca(2+) release induced by MP was further increased in the presence of caffeine, showing that the MP binding site is different from that of caffeine on Ca(2+) release channels. We succeeded in the synthesis of (125)I-[Tyr(3)]MP with a high specific activity. (125)I-[Tyr(3)]MP bound specifically to heavy fraction of sarcoplasmic reticulum with a K(d) value of 4.0 microM and a B(max) value of 3.0 nmol/mg. Furthermore, (125)I-[Tyr(3)]MP specifically cross-linked to the 97 kDa protein without direct binding to ryanodine receptor. The protein was not triadin or Ca(2+)-pump, because antitriadin antibody and anti-Ca(2+)-pump antibody did not immunoprecipitate the protein. These results suggest that the 97 kDa MP-binding protein may have an important role in the excitation-contraction coupling of skeletal muscle. PMID- 10825396 TI - Inactivation studies of acetylcholinesterase with phenylmethylsulfonyl fluoride. AB - Acetylcholinesterase (AChE), a serine hydrolase, is potentially susceptible to inactivation by phenylmethylsulfonyl fluoride (PMSF) and benzenesulfonyl fluoride (BSF). Although BSF inhibits both mouse and Torpedo californica AChE, PMSF does not react measurably with the T. californica enzyme. To understand the residue changes responsible for the change in reactivity, we studied the inactivation of wild-type T. californica and mouse AChE and mutants of both by BSF and PMSF both in the presence and absence of substrate. The enzymes investigated were wild-type mouse AChE, wild-type T. californica AChE, wild-type mouse butyrylcholinesterase, mouse Y330F, Y330A, F288L, and F290I, and the double mutant T. californica F288L/F290V (all mutants given T. californica numbering). Inactivation rate constants for T. californica AChE confirmed previous reports that this enzyme is not inactivated by PMSF. Wild-type mouse AChE and mouse mutants Y330F and Y330A all had similar inactivation rate constants with PMSF, implying that the difference between mouse and T. californica AChE at position 330 is not responsible for their differing PMSF sensitivities. In addition, butyrylcholinesterase and mouse AChE mutants F288L and F290I had increased rate constants ( approximately 14 fold) over those of wild-type mouse AChE, indicating that these residues may be responsible for the increased sensitivity to inactivation by PMSF of butyrylcholinesterase. The double mutant T. californica AChE F288L/F290V had a rate constant nearly identical with the rate constant for the F288L and F290I mouse mutant AChEs, representing an increase of approximately 4000-fold over the T. californica wild-type enzyme. It remains unclear why these two positions have more importance for T. californica AChE than for mouse AChE. PMID- 10825397 TI - Importance of phenylalanine 107 in agonist recognition by the 5 hydroxytryptamine(3A) receptor. AB - The 5-hydroxytryptamine (5-HT)(3) receptor is a member of the ligand-gated ion channel receptor family with significant homology to the nicotinic acetylcholine, gamma-aminobutyric acid(A), and glycine receptors. In this receptor class, the agonist binding site is formed by parts of the extracellular amino-terminal region. This study examines the effects of altering phenylalanine 107 (F107) of the 5-HT(3AL) subunit, obtained from NG108-15 cells, using site-directed mutagenesis. The wild-type (WT) and mutant receptors were expressed in HEK 293 cells and characterized using both whole-cell patch-clamp and radioligand binding. The tyrosine mutant F107Y exhibits a significantly lower affinity for the agonist 5-HT (K(i) = 203 versus 15.6 nM) and an increase of similar magnitude in the EC(50) value (10.6 versus 1.2 microM) compared with WT. The activation kinetics of the maximal currents generated by 5-HT with this mutant were markedly slower than those of the WT receptor, but application of supramaximal concentrations of the agonist markedly decreased the time to half-peak. The asparagine mutant F107N displayed a significantly higher affinity for 5-HT than the WT receptor (1.62 versus 15.6 nM), which was mirrored in direction and magnitude by changes in the EC(50) value for this agonist (0.2 versus 1.2 microM). In contrast to the WT receptor, the mutant F107N was activated by acetylcholine (EC(50) = 260 microM). The response to acetylcholine was blocked by the 5-HT(3) receptor antagonist renzapride with a similar IC(50) value as that determined against currents generated by 5-HT in the WT receptor. These data suggest that F107 is an important determinant of agonist recognition at the 5 HT(3) receptor. PMID- 10825398 TI - Nucleotide modulation of pinacidil stimulation of the cloned K(ATP) channel Kir6.2/SUR2A. AB - ATP-sensitive K(+) channels are the target for K(+) channel openers such as pinacidil. These channels are formed from pore-forming Kir6. 2 and regulatory sulfonylurea receptor (SUR) subunits. Pinacidil activates channels containing SUR2A (heart, skeletal muscle), but not those containing SUR1 (beta cells). Surprisingly, binding of the pinacidil analog [(3)H]P1075 is dependent on added nucleotides, yet in electrophysiological studies, pinacidil is effective in the absence of intracellular nucleotides. To determine the reason for this anomaly, we examined the functional interactions between pinacidil (or P1075) and nucleotides by expressing cloned Kir6. 2/SUR2A channels in Xenopus laevis oocytes. Both pinacidil and P1075 activated macroscopic Kir6.2/SUR2A currents in the absence of added nucleotide, but the presence of intracellular ATP or ADP slowed the off-rate of the response. Mutation of the Walker A lysine in a single nucleotide binding domain (NBD) of SUR2A (K707A in NBD1, K1348A in NBD2), abolished this action of nucleotide. The K1348A mutation prevented stimulation by MgADP but had little effect on the amplitude of the pinacidil response. In contrast, Kir6.2/SUR2A-K707A currents were activated by MgADP, but only responded to pinacidil in the presence of Mg-nucleotide. Off-rates in the absence (or presence) of nucleotide were slower for the pinacidil analog P1075 than for pinacidil, consistent with the higher affinity of P1075. We suggest that slowing of P1075 dissociation by nucleotide enables binding to be detected. PMID- 10825399 TI - Allopregnanolone synthesis in cerebellar granule cells: roles in regulation of GABA(A) receptor expression and function during progesterone treatment and withdrawal. AB - Rat cerebellar granule cells were cultured for 5 days with progesterone, resulting in the conversion of progesterone to allopregnanolone, a potent and efficacious modulator of gamma-aminobutyric acid (GABA) type-A receptors, as well as in decreases in the abundance of GABA(A) receptor alpha(1), alpha(3), alpha(5), and gamma(2) subunit mRNAs. These effects were accompanied by decreases in the efficacies of diazepam and the beta-carboline DMCM with regard to modulation of GABA-evoked Cl(-) currents. Withdrawal from such progesterone treatment resulted in a rapid and selective increase in the abundance of the GABA(A) alpha(4) subunit mRNA that was associated with a restoration of receptor sensitivity to the negative modulatory action of DMCM, a positive receptor response to flumazenil, and continued reduced responsiveness of receptors to diazepam. Prevention of allopregnanolone synthesis by the 5alpha-reductase inhibitor finasteride also prevented the changes in both GABA(A) receptor gene expression and receptor function elicited by progesterone treatment and withdrawal. PMID- 10825400 TI - Prospects for effective treatment of AL amyloidosis? PMID- 10825401 TI - Pulmonary embolism-an update on thrombolytic therapy. PMID- 10825402 TI - Curative hepatorenal transplantation in systemic amyloidosis caused by the Glu526Val fibrinogen alpha-chain variant in an English family. AB - A 53-year-old English woman who had been thought to have systemic monoclonal immunoglobulin light chain (AL) amyloidosis was investigated further because of her unusually long 17-year history and a suggestion of renal disease in the family. She was found to have the Glu526Val fibrinogen alpha-chain variant that causes autosomal dominant hereditary systemic amyloidosis. This has not previously been described in a British family. The mutant gene was associated with the same haplotype as in all other reported cases, suggesting a common founder. The patient had already received a renal transplant, but the graft failed within 6 years due to amyloid deposition. Progressive hepatic amyloidosis eventually caused liver failure, although the function of other organs was well preserved. She therefore received hepatic and renal transplants to replace the failed organs and the hepatic source of the amyloidogenic variant fibrinogen. Three years later she is completely well and has no amyloid deposits identifiable by serum amyloid P component scintigraphy. This is the first detailed report of hepatic transplantation for liver failure caused by amyloidosis of any type. The substantial follow-up suggests that fibrinogen alpha-chain amyloidosis is one of the inherited metabolic diseases that can be cured by liver transplantation. The mutation underlying Glu526Val fibrinogen alpha-chain amyloidosis is incompletely penetrant and has a variable phenotype that can clinically mimic AL amyloidosis. Hereditary fibrinogen amyloidosis may be more prevalent than previously suspected and, since AL amyloid is sometimes a diagnosis of exclusion, genotyping for other amyloidogenic proteins is mandatory in all cases in which the amyloid fibrils cannot be positively identified as AL. PMID- 10825403 TI - Computer-assisted venous occlusion plethysmography in the diagnosis of acute deep venous thrombosis. AB - Suspicion of deep venous thrombosis (DVT) is a common reason for acute medical admission. The clinical diagnosis is difficult, and thus significant numbers are investigated and found to be normal. Provision of 24-h radiology is costly, and there may be a delay in investigation. We assessed computer-assisted venous occlusion plethysmography as a screening test for DVT, compared with standard radiology. The test has the advantage of being performed on the ward and if reliable would significantly reduce the number of radiological investigations required. We enrolled 215 consecutive patients presenting with the possible diagnosis of DVT, of whom 144 had technically adequate plethysmography results. Plethysmography had a sensitivity of 96% (95%CI 88-99%) and a negative predictive value of 97% (95%CI 91-99%). Patients excluded because of technically inadequate results were older (by a mean 7 years, p=0. 003). Computer-assisted venous occlusion plethysmography is a non-invasive method of rapidly screening for DVT which may be safely used as an initial screening test. The test is less useful in older patients, or patients unable to keep still for a period of 2 min. PMID- 10825404 TI - Fatigue in Cambodia veterans. AB - In 1992 and 1993, Dutch military personnel were deployed in the peace operation UNTAC in Cambodia. Since returning, Cambodia veterans have reported health complaints which they perceive to be related to their service. Their symptoms strikingly resemble health problems reported by Gulf War veterans. Four years post-return, a cross-sectional survey on health symptoms in Cambodia veterans was initiated. Questionnaires were sent to all Cambodia veterans and four comparison groups. Forgetfulness, difficulty concentrating and fatigue were the symptoms most commonly endorsed. An operational case definition was constructed using a validated fatigue severity questionnaire. Cases were not uniquely found in Cambodia veterans (17%). In Rwanda and Bosnia veterans, respectively, 28% and 11% also met our case definition. Fatigue severity level was predicted by pre mission, during-mission and post-mission variables, of which retrospective recollection of side-effects of vaccines and causal attributions also have been shown to be relevant in studies on Gulf-related illness. PMID- 10825405 TI - The burden of alcohol misuse on an inner-city general hospital. AB - Alcohol consumption in the UK has been increasing steadily. We prospectively studied the burden on hospital services caused by overt alcohol misuse, in an inner-city hospital in north-west England. All Accident & Emergency (A&E) patients were assessed to determine whether their hospital attendance was alcohol related, and whether this resulted in admission and/or generated new out-patient appointments. Over 2 months, 1915 patients attended A&E with alcohol-related problems, accounting for 12% of attendances; 50% were aged 18-39 years, and acute alcohol intoxication was the commonest presenting complaint. Overall, 6.2% of all hospital admissions were due to alcohol-related problems. Over 2800 new out patient visits were likely to have been generated over an 18-month period from initial attendance with an alcohol-related problem, mostly for orthopaedic clinics. The burden placed by overt alcohol-related problems on hospitals is enormous, both in terms of the emergency and out-patient services. The implementation of education, screening and intervention strategies in A&E departments, and employment of key trained personnel, should be considered, to optimize the clinical management of these patients. PMID- 10825406 TI - Renal involvement in primary Sjogren's syndrome. AB - Renal involvement was evaluated in 62 patients with primary Sjogren's syndrome, classified according to criteria proposed by The European Classification Criteria Group. Urine concentration capacity was tested using intranasal 1-desamino-8-D arginine-vasopressin. For patients with urine pH>5.5 without metabolic acidosis (n=28), an acidification test with ammonium chloride was performed. Urinary citrate, albumin, NAG, ALP and beta2-microglobulin were measured and creatinine clearance was calculated. Maximum urine concentration capacity and creatinine clearance were reduced in 13 (21%). Albumin excretion was >30 microg/min in only one patient (1.6%). Seven patients (11.3%) had complete or incomplete distal renal tubular acidosis (dRTA), four had reduced creatinine clearance and five had reduced maximum urine concentration capacity. The ratio of citrate/creatinine in spot urine was below the 2.5 percentile in all patients with complete or incomplete dRTA. The prevalence of dRTA was lower than in previous studies. There were also few patients with signs of glomerular disease (1.6%). The use of citrate:creatinine ratio in spot urine can be a helpful method in identifying patients with complete or incomplete dRTA. PMID- 10825407 TI - New therapies for heart failure: is thalidomide the answer? AB - The syndrome of advanced heart failure is associated with considerable morbidity and mortality. Ideas about the reasons for the progressive nature of the heart failure syndrome have changed over the years, with the initial view that progression was principally due to pump failure (the 'haemodynamic' hypothesis), giving way to more modern views, which implicate neuro-endocrine activation (including catecholamine excess, renin-angiotensin system activation, etc.). More recently, an excess of inflammatory cytokines has been found in advanced heart failure and implicated in the progression of the disease. Amongst the cytokines found, TNF-alpha seems to be particularly important. The principle therapeutic action of thalidomide appears to be reduction of TNF-alpha levels. We therefore suggest that there may be a role for thalidomide, or its derivatives, in the management of advanced heart failure. PMID- 10825408 TI - A hypothesis regarding the origin and spread of the cystic fibrosis mutation deltaF508. PMID- 10825409 TI - Migraine and stroke in young women. PMID- 10825410 TI - Potassium excretion indices in the diagnostic approach to hypokalaemia. PMID- 10825411 TI - The secretory response of the rat colon to the flavonol quercetin is dependent on Ca2+-calmodulin. AB - The dietary flavonol quercetin induces chloride secretion in rat intestine. To clarify the underlying mechanisms, experiments were performed in Ussing chambers with tissue from rat proximal and distal colon. Quercetin induced an increase in short-circuit current (Isc), which was largely independent of submucosal neurons, as it was not affected by the neurotoxin tetrodotoxin. The effect of quercetin was blocked by the calmodulin antagonists trifluoperazine and ophiobolin A and was diminished by a blocker of Ca2+ release from intracellular stores (TMB-8), whereas the muscarinic receptor antagonist atropine was ineffective. The quercetin-induced Isc was abolished in Ca2+-free solution. The flavonol was able to further increase Isc after maximal stimulation of the cAMP pathway by forskolin. The Isc increase by the flavonol was differently affected by two analogous phosphodiesterase inhibitors. Whereas 3-isobutyl-1-methylxanthine (IBMX) antagonized the effect of quercetin, 8-methoxymethyl-IBMX had no effect. Both phosphodiesterase inhibitors similarly influenced the Isc increase induced by forskolin. These results indicate that the chloride secretion induced by quercetin in rat colon depends on Ca2+ and calmodulin. The cAMP pathway and inhibition of phosphodiesterase appear not to be responsible for the secretory activity of the flavonol. PMID- 10825412 TI - Activation of human leukocytes by acute hypoxia. AB - Exposure of human leukocytes to 12% and 6% O(2) in vitro resulted in striking ultrastructural and morphological changes. These changes included the appearance of crater-like holes, a reduction in granular size and disruption of the cellular membrane. The implications of these findings is discussed. PMID- 10825413 TI - Alteration in alpha- and beta-adrenoceptor profile of rabbit knee joint blood vessels due to acute inflammation. AB - Experiments were performed to investigate the nature of - and -adrenoceptors in blood vessels supplying the posterior capsule of the acutely inflamed rabbit knee joint, and results were compared to findings from previous experiments on the normal joint, to assess any alteration which may occur in the adrenoceptor profile due to the inflammation process. Electrical stimulation of the posterior articular nerve resulted in vasoconstriction which was reversed to vasodilatation by phentolamine and yohimbine. The dose-response curves to close intra-arterial injection of -adrenoceptor agonists showed a rank-order potency of: adrenaline = phenylephrine = clonidine. The adrenaline dose-response curve was shifted to the right by administration of antagonists with a rank-order potency of: phentolamine = yohimbine = prazosin. At this stage of the experiments there was an equal response of 1- and 2-adrenoceptors in blood vessels of the acutely inflamed rabbit knee joint. In another group of animals the neurally mediated vasodilatation, which appeared after administration of phentolamine, was completely blocked by propranolol, and was reduced by about 50 % by atenolol. The dose-response curves to close intra-arterial injection of -adrenoceptor agonists showed a rank-order potency of: isoprenaline > salbutamol = dobutamine. The isoprenaline dose-response curve was shifted to the right by administration of antagonists with a rank-order potency of: propranolol > atenolol. These experiments also showed an almost equal response of 1- and 2-adrenoceptors in blood vessels of the acutely inflamed rabbit knee joint. Overall, compared to previous experiments on the normal joint in which 2- and 1-adrenoceptor responses predominated, acute inflammation resulted in a shift from 2- towards 1- and from 1- towards 2-adrenoceptor responses. PMID- 10825414 TI - Response of isolated ruminant mammary arteries to the long R3 analogue of insulin like growth factor I. AB - Isolated mammary arteries from ruminants were used in a conventional organ bath system. Acetylcholine relaxed bovine but not ovine mammary arteries; both types responded to sodium nitroprusside. Noradrenaline (NA) caused a dose-dependent increase in generated tension. An analogue of insulin-like growth factor I (long R3-IGF-I) caused a rightward shift in the NA response curve in bovine vessels with intact endothelium (P < 0.02), and also in sheep arteries (P < 0.01). In bovine vessels, this effect was abolished when the endothelium was removed. The effect of long R3-IGF-I in bovine vessels was abolished by N -nitro-L-arginine methyl ester (L-NAME) an inhibitor of nitric oxide synthase, suggesting the effect of IGF-I on mammary arteries in vitro requires NO generation. PMID- 10825415 TI - Immunoglobulin A secretion into saliva during dual sympathetic and parasympathetic nerve stimulation of rat submandibular glands. AB - Salivary secretion of proteins from rat submandibular glands was studied using graded stimulation of the parasympathetic nerve in isolation, and then at a fixed rate in combination with graded sympathetic nerve stimulation. Increasing the frequency of parasympathetic nerve stimulation per se caused a gradual increase in the secretion of peroxidase (from acini) but only small increases in proteinase (from ductal cells) and IgA outputs. Dual stimulations, with an increasing frequency of sympathetic nerve stimulation on a background of low frequency parasympathetic nerve stimulation, showed that maximal acinar secretion of peroxidase required only a low frequency of additional sympathetic stimulation, whereas ductal secretion of kallikrein was greatest with the highest frequency of additional sympathetic stimulation (20 Hz in bursts). IgA secretion also required high frequency additional sympathetic stimulation in bursts for greatest output. Although a synergism occurred with parasympathetic plus sympathetic nerve stimulation for the secretion of both peroxidase and kallikrein it was not evident for the secretion of IgA. This presumably reflects a difference for exocytosis of proteins stored in granules (e.g. peroxidase and kallikrein) compared to those proteins continuously transported across the plasma membrane in vesicles by transcytosis. This work confirms that vesicular movement of secretory IgA can be increased by both parasympathetic and sympathetic nerve stimulation, but the frequency parameters differ for each nerve. PMID- 10825416 TI - Renin and heart rate responses to haemorrhage are age dependent in conscious lambs. AB - The present experiments were carried out in conscious lambs (1-2 weeks old, n = 9) and older sheep (11-12 weeks old, n = 11) to determine whether the cardiovascular and endocrine responses to 0, 10 and 20 % haemorrhage were developmentally regulated. The major novel finding of our study is that throughout the first 3 months of postnatal life, there is a similar decrease in mean arterial pressure and a similar restoration of pressure to pre-haemorrhage levels, for the same degree of blood loss, yet the mechanisms used to restore pressure appear to be age dependent as follows. In lambs, but not in older sheep, heart rate increased for 1 h after 20% haemorrhage. Activation of the renin angiotensin system was also greater and more prolonged in lambs than in older sheep following haemorrhage, and occurred at a lesser degree of blood loss. Plasma arginine vasopressin responses to haemorrhage were, however, similar in both age groups. These data provide new information that some of the mechanisms used to restore arterial pressure following blood volume depletion appear to be age dependent. PMID- 10825417 TI - Power output of fast and slow skeletal muscles of mdx (dystrophic) and control mice after clenbuterol treatment. AB - The mdx mouse is the most commonly used animal model for Duchenne muscular dystrophy. We tested the null hypothesis that 20 weeks of clenbuterol treatment ( approximately 2 mg kg-1 day-1) of mdx and control mice would have no effect on the absolute and specific force (Po, kN m-2) and absolute and normalised power output (W kg-1) of extensor digitorum longus (EDL) and soleus muscles. For mdx and control mice, clenbuterol treatment produced modest increases in the mass of the two muscles but did not increase absolute or specific force or normalised power output. For absolute power output, only the EDL muscles of mdx mice showed a difference following treatment, with the power output of treated mice being 118 % that of the untreated mice. The modest effects of clenbuterol treatment on the dynamic properties of skeletal muscle provide little support for any improvement in muscle function for the dystrophic condition. PMID- 10825418 TI - A respiratory drive in addition to the increase in CO(2) production at raised body temperature in rats. AB - Mammals that use the ventilatory system as the principal means of increasing heat loss, i.e. that pant, show two fundamental changes in the control of breathing at raised temperatures. First, alveolar ventilation increases by more than, rather than in proportion to, the increase in CO2 production. Second, hypocapnia no longer causes apnoea. Rats do not use the ventilatory system as the principal means of increasing heat loss, so we have investigated whether rats also show these two changes at raised temperatures. Breathing was detected from diaphragmatic electromyogram (EMG) activity. Anaesthesia and hyperoxia were used to minimise behavioural and hypoxic drives to ventilation and arterial PCO2 (Pa,CO2) was controlled using mechanical ventilation. At 36.6 +/- 0.1 >C, breathing was absent as long as Pa,CO2 was held below a threshold level of 32.9 +/- 0.7 mm Hg (n = 14) under steady-state conditions. When body temperature in rats was raised above 37 >C, both fundamental changes in the control of breathing became apparent. First, at 39 >C the mean Pa,CO2 level during spontaneous breathing (39.6 +/- 5.4 mm Hg, n = 4) fell by 3.9 +/- 1.4 mm Hg (P < 0.05, Student's paired t test). Second, at 39.9 +/- 0.1 >C breathing was present when mean Pa,CO2 levels were only 18.2 +/- 1.5 mm Hg (n = 14), the lowest mean Pa,CO2 level we could achieve with mechanical ventilation. We calculate, however, that at 39.9 >C, the drive to breathe from the increased CO2 production alone would not sustain breathing below a Pa,CO2 level of 27.8 +/- 1.4 mm Hg (n = 13). In rats at raised body temperatures therefore a respiratory drive exists that is in addition to that related to the increase in CO2 production. PMID- 10825419 TI - Thermoregulation in winter swimmers and physiological significance of human catecholamine thermogenesis. AB - Thermoregulation in control subjects and cold-adapted winter swimmers was examined during 1 h of cold water immersion (13 C). It was found that the thermoregulatory functions of winter swimmers differ from those of non-cold adapted subjects. As evident from the relationship between rectal temperature and the magnitude of cold thermogenesis, in controls a significant part of cold thermogenesis during the early phase of cooling was induced by changes in peripheral temperature input, while in the late phase of cooling it was the central temperature input which was mainly engaged in induction of cold thermogenesis. In winter swimmers the magnitude of cold thermogenesis was solely related to changes in rectal temperature, indicating the predominance of the central temperature input in activation of heat production mechanisms. The thermoregulatory threshold for induction of cold thermogenesis was lowered (by 0.34 C), but the apparent hypothalamic thermosensitivity was the same as in non cold-adapted subjects. These differences are indicative of adaptation of thermoregulatory control centres. Additionally, the activity of thermoregulatory effectors was also changed. Shivering was induced later during cooling (after 40 min) in winter swimmers than in controls, which suggests an important participation of non-shivering thermogenesis in the early thermogenic response. Winter swimmers also showed bradycardia and a greater reduction in plasma volume during cooling. The data indirectly indicate restriction of heat loss from the body. Only a non-significant increase in quantity of subcutaneous fat was observed in winter swimmers. Thus, winter swimmers were able to survive a significantly greater temperature gradient between body and environment than non cold-adapted subjects by modifying the sensory functions of hypothalamic thermoregulatory centres to lower heat loss and produce less heat during cold exposure. Additionally, the capacity of the total cold thermogenesis due to potentiation of non-shivering heat production was also increased. Heat produced due to thermogenic action of adrenaline may represent more than a quarter of the total cold thermogenesis. In conclusion, the data suggest that winter swimmers exhibit metabolic, hypothermic and insulative types of cold adaptation. PMID- 10825420 TI - Heart rate and its variability change after the menopause. AB - Resting heart rate and heart rate variability of 33 postmenopausal women were compared with those of 50 premenopausal women of comparable activity level, none of whom had used hormone replacement therapy. Heart rate was measured as the mean of at least 600 consecutive R-R intervals obtained from electrocardiograph (ECG) records, and its variability as the standard deviation of these intervals. Activity levels were assessed by a scale modified from the Allied Dunbar National Fitness Survey (1992). There was a significant reduction in both mean R-R interval and the standard deviation in the postmenopausal women who had experienced their last menstrual period (LMP) 1 year or more prior to the observations being made, but no observable changes during the first year post menopause. PMID- 10825421 TI - Effects of lung volume on parasternal pressure-generating capacity in dogs. AB - Previous studies have suggested that the optimum length for force generation of the parasternal intercostal (PS) muscles is well above functional residual capacity (FRC). We further explored this issue by examining the pressure generating capacity of the PS muscles as a function of lung volume in anaesthetized dogs. Upper thoracic spinal cord stimulation (SCS) was used to electrically activate the PS muscles. Changes in airway pressure and parasternal resting length (LR) during airway occlusion were monitored over a wide range of lung volumes during SCS. To assess the effects of parasternal contraction alone, SCS was performed following phrenicotomy and section of the external intercostal, levator costae and triangularis sterni muscles. With increasing lung volume, there were progressive decrements in the capacity of the PS muscles to produce changes in airway pressure. The relationship between PS pressure generation and lung volume was similar to a previous comparable assessment of the external intercostal muscles. The PS muscles shortened during passive inflation and also shortened further (by > 20 % of LR) during SCS. Total shortening (passive plus active) increased progressively with increasing lung volume. Our results indicate that the capacity of the PS muscles to produce changes in airway pressure (a) falls progressively with increasing lung volume and (b) is similar to that of the external intercostal muscles. We speculate that the fall in PS pressure generating capacity is related, in part, to progressive reductions in end inspiratory length. PMID- 10825422 TI - The off-transient pulmonary oxygen uptake (VO(2)) kinetics following attainment of a particular VO(2) during heavy-intensity exercise in humans. AB - The oxygen uptake response to moderate-intensity exercise (i.e. < anaerobic threshold (an)) has been characterised with a gain (i.e. response amplitude per increment of work rate) and time constant that do not vary appreciably at different work rates or between the on- and off-transients. Above an, the response becomes more complex with an early component that typically projects to a value that has a gain similar to that of the < an response, but which is supplemented by the addition of a delayed slow kinetic component. We therefore established a constant target VO2 (VO21) for each subject such that with different imposed work rates the contribution to VO21 from the slow phase varied over a wide range. Work rates were chosen so that VO21 was attained at 2-24 min. Five subjects (aged 21-58 years) cycled at four to five different work rates. VO2 was measured breath-by-breath, at VO21 the work rate was abruptly reduced and the subject recovered by cycling unloaded for 15 min. Unlike the on-transient, for which the slow component shows a long delay, the off-transient was best fitted as two simultaneous exponential components. The slower off-transient component had a small amplitude and long time constant, but did not differ significantly among the various tests. The off-transient kinetics for VO2 therefore was independent of the magnitude of the contribution to the slow phase from the on-transient kinetics. PMID- 10825423 TI - Simultaneous monitoring of vascular contractility, intracellular pH and intracellular calcium in isolated rat mesenteric arteries; effects of weak bases. AB - We report the first simultaneous measurements of pHi, [Ca2+]i and tension, upon alteration of pHi, in isolated rat mesenteric arteries loaded with both carboxy SNARF and indo-1. In these vessels (pre-contracted by 30 mM KCl) alterations of pHi, by addition and subsequent washout of weak bases, produced complicated effects on tone. Although the changes in contractility did not mirror the changes in pHi they were, at all times, accompanied by parallel changes in [Ca2+]i. PMID- 10825424 TI - Solution structure of the loops of bacteriorhodopsin closely resembles the crystal structure. AB - Bacteriorhodopsin is one of very few transmembrane proteins for which high resolution structures have been solved. The structure shows a bundle of seven helices connected by six turns. Some turns in proteins are stabilized by short range interactions and can behave as small domains. These observations suggest that peptides containing the sequence of the turns in a membrane protein such as bacteriorhodopsin may form stable turn structures in solution. To test this hypothesis, we determined the solution structure of three peptides each containing the sequence of one of the turns in bacteriorhodopsin. The solution structures of the peptides closely resemble the structures of the corresponding turns in the high resolution structures of the intact protein. PMID- 10825425 TI - The reduced folate carrier in L1210 murine leukemia cells is a 58-kDa protein. AB - The reduced folate carrier (RFC1) is a major transporter for both natural reduced folates and antifolate chemotherapeutics. Using polyclonal antibodies targeted to epitopes at the loop between the sixth and seventh predicted transmembrane domains or the distal C-terminus, we were able to demonstrate by Western blot analysis that the molecular size of RFC1 expressed in murine leukemia L1210 cells is 58 kDa as predicted by the open reading frame of its cDNA. 46- and 38-kDa proteins detected only in plasma membrane preparations were proteolytic degradation products that appeared during membrane preparation or treatment with the conventional SDS-PAGE loading buffer. These data resolve discrepancies reported previously for the molecular size of RFC1. PMID- 10825426 TI - Cationic liposome and plasmid DNA complexes formed in serum-free medium under optimum transfection condition are negatively charged. AB - In medium where in vitro transfection is routinely performed, DC-chol liposomes alone were nearly neutral, whereas the DC-chol liposome/DNA complexes were largely negatively charged which changed only slightly at all [liposome]/[DNA] ratios (zeta=-27.1 to -21.8 mV). Three other commercial transfection reagents, Lipofectin(R), LipofectAMINE 2000, and SuperFect, were also largely negatively charged when complexed with DNA. The aggregation of liposomes in medium was prevented by the addition of DNA. Incubation of the complexes in medium did not change their size, charge or lipofection activity for 30 min. These results suggest that, in medium, the liposome/DNA complexes were formed at the time of mixing with negative charges. PMID- 10825427 TI - Identification of type 1 IP(3) receptors in the rat kidney and their modulation by immobilization stress. AB - Inositol 1,4,5-trisphosphate receptor (IP(3)-receptor) is a calcium channel, transporting calcium from intracellular stores to the cytoplasm. In kidney, IP(3) receptors are involved in the signal transduction of various hormones. In our work we studied the effect of immobilization stress on the IP(3)-receptor's protein content in renal cortex and the medulla of normotensive and hypertensive rats. We detected both mRNA and type 1 IP(3)-receptor protein in medulla, but not in renal cortex. We found that this receptor was approximately twice as abundant in normotensive as in genetically hypertensive rat kidney. Immobilization stress decreased the amount of type 1 IP(3)-receptor in the renal medulla of normotensive rats approximately five times, while no effect due to single and/or repeated stress was observed in the renal medulla of spontaneously hypertensive rats. The results indicate that expression of type 1 IP(3)-receptor in renal medulla is modulated by hypertension and immobilization stress. PMID- 10825428 TI - Crystallization of the isobutylphosphocholine-cholesterol-isobutanol (1:3:3) complex and its investigation by X-ray analysis: interaction of phopholipid headgroups with cholesterol. AB - A crystal complex consisting of the isobutyl analog of phosphatidylcholine (PC) (isobutylphosphocholine), cholesterol, and isobutanol with molecular ratio 1:3:3 was obtained and investigated by means of X-ray analysis. The complex was shown to correspond to the monoclinic system (sp. gr. P2(1)): a=16.994(10), b=11.314(7), c=28.164(15), beta=104.07(3), V=5252.63 A(3), Z=2, D(calc)=1.0273 g/cm(3). The isobutylphosphocholine molecule is the key component of the complex. Pairs of hydrogen bonds are formed between the (-delta)O-P-O(delta-) group of the isobutylphosphocholine molecule and C-OH groups of two cholesterol and two isobutanol molecules. The third molecules of cholesterol and isobutanol are H bonded with the (-delta)O-P-O(delta-) group of the isobutylphosphocholine molecule via C-OH groups of isobutanol and cholesterol, respectively. The crystal structure is built up by translation of the complex in multiplicate along the two fold axis in the direction of axis b. It contains bands formed by isobutylphosphocholine molecules alternately changing their direction. They are fixed by virtue of two zones of electrostatic interactions of the type (-delta)O P-O(delta-)ellipsis(+)N(CH(3))(3) and are more or less parallel to the bc plane. The structure also contains three-layer domains formed by cholesterol molecules perpendicular to isobutylphosphocholine bands. In the direction of the c-axis isobutylphosphocholine bands alternate with the layers of cholesterol molecules herewith reproducing repeated blocks. The obtained structure is compared with that of crystals of phospholipids and cholesterol and its derivatives. PMID- 10825429 TI - Investigations of spectrin-lipid interactions using fluoresceinphosphatidylethanolamine as a membrane probe. AB - The binding of human erythrocyte spectrin to large unilamellar vesicles (LUVET) formed by the extrusion technique has been studied using fluoresceinphosphatidylethanolamine (FPE) as a reporter of electrostatic membrane potential. Spectrin aliquots were added to a suspension of FPE-labelled LUVETs to elucidate both the type of charge involved and the dissociation constants for spectrin binding to various lipids. All binding experiments showed serial increases in FPE fluorescence intensity upon serial additions of spectrin, indicative of increasing positive charge at the membrane surface. This proves for the first time that although exhibiting an overall net negative charge, spectrin binds to lipid surfaces by presenting positive charges to the lipid surface. Binding curves were obtained from the change in fluorescence intensity upon each spectrin addition and analysed to determine dissociation constants. A K(d) of 0.14+/-0.12 microM was found for spectrin binding to FPE-labelled phosphatidylcholine/phosphatidylserine (PC/PS) LUVETs at 22 degrees C in high salt conditions. A similar K(d) of 0.17+/-0.11 microM was obtained for spectrin binding to neutral LUVETs composed of PC. However, binding was found to be much weaker for PC/PS LUVETs under low salt conditions with a K(d) of 1.22+/-0.48 microM. PMID- 10825430 TI - A synthetic peptide based on a glycine-gated chloride channel induces a novel chloride conductance in isolated epithelial cells. AB - CK(4)-M2GlyR, an aqueous soluble peptide derived from the transmembrane M2 segment of the glycine-gated Cl(-) channel found in postsynaptic membranes of the central nervous system, has previously been shown to increase transepithelial Cl( ) and fluid secretion of epithelial monolayers. The goal of this study was to determine whether CK(4)-M2GlyR exerts these effects via formation of a novel chloride conductance pathway, modulation of endogenous chloride channel activity, or a combination of these effects. Ionic currents were recorded from isolated epithelial cells before and after treatment with the peptide using the whole-cell configuration of the patch-clamp technique. CK(4)-M2GlyR increased whole-cell Cl( ) currents in all epithelial cell lines that were studied, including: Madin-Darby canine kidney cells, a human colonic epithelial cell line (T84), and airway epithelial cells derived from a human cystic fibrosis patient (IB3-1). No evidence was found for modulation of endogenous Cl(-) channels by CK(4)-M2GlyR based on both the electrophysiological properties of the observed currents and the pharmacological profile of the CK(4)-M2GlyR-induced current. These results suggest that CK(4)-M2GlyR increases Cl(-) permeability in epithelial cells directly, by forming a distinct conduction pathway in cell membranes. PMID- 10825431 TI - Involvement of rBAT in Na(+)-dependent and -independent transport of the neurotransmitter candidate L-DOPA in Xenopus laevis oocytes injected with rabbit small intestinal epithelium poly A(+) RNA. AB - Although L-3,4-dihydroxyphenylalanine (L-DOPA) is claimed to be a neurotransmitter in the central nervous system (CNS), receptor or transporter molecules for L-DOPA have not been determined. In an attempt to identify a transporter for L-DOPA, we examined whether or not an active and high affinity L DOPA transport system is expressed in Xenopus laevis oocytes injected with poly A(+) RNA prepared from several tissues. Among the poly A(+) RNAs tested, rabbit intestinal epithelium poly A(+) RNA gave the highest transport activity for L [(14)C]DOPA in the oocytes. The uptake was approximately five times higher than that of water-injected oocytes, and was partially Na(+)-dependent. L-Tyrosine, L phenylalanine, L-leucine and L-lysine inhibited this transport activity, whereas D-DOPA, dopamine, glutamate and L-DOPA cyclohexylester, an L-DOPA antagonist did not affect this transport. Coinjection of an antisense cRNA, as well as oligonucleotide complementary to rabbit rBAT (NBAT) cDNA almost completely inhibited the uptake of L-[(14)C]DOPA in the oocytes. On the other hand, an antisense cRNA of rabbit 4F2hc barely affected this L-[(14)C]DOPA uptake activity. rBAT was thus responsible for the L-[(14)C]DOPA uptake activity expressed in X. laevis oocytes injected with poly A(+) RNA from rabbit intestinal epithelium. As rBAT is localized at the target regions of L-DOPA in the CNS, rBAT might be one of the components involved in L-DOPAergic neurotransmission. PMID- 10825432 TI - Targeting of endothelial KDR receptors with 3G2 immunoliposomes in vitro. AB - Immunoliposomes (IL) containing anti-angiogenic drugs directed selectively to the easily accessible kinase insert domain containing receptor (KDR) vascular endothelial growth factor (VEGF), which is predominantly expressed on tumour vessels are a promising tool to inhibit tumour angiogenesis. To explore this strategy, we have prepared fluorescent-labelled IL presenting antibodies against the KDR receptor (3G2) on their surface. 3G2-IL were composed of egg phosphatidylcholine and cholesterol (6:4), containing 2 mol% of the new thiol reactive linker lipid O-(3-cholesteryloxycarbonyl)propionyl-O'-m-maleimido benzoyl tetraethylene glycol. Specific binding of 3G2-IL to immobilised recombinant KDR was used to show the maintenance of sufficient immunoreactivity of 3G2 antibodies upon the coupling procedure. 3G2-IL bound to Chinese hamster ovarian (CHO) cells stably transfected to overexpress KDR to a five times higher amount as compared to mock-transfected CHO cells. Subsequently, specific binding of 3G2-IL to KDR could also be demonstrated on KDR expressing cells, human umbilical vein endothelial cells and human microvascular endothelial cells, whereas only low binding of 3G2-IL to NIH-3T3 mouse fibroblast cells, which do not express KDR, was found. The binding of 3G2-IL to KDR receptors could not be blocked by VEGF, suggesting that the binding site for VEGF is not identical with the epitope recognised by 3G2. We could demonstrate that 3G2-IL is able to bind in vitro even in the presence of high levels of VEGF. PMID- 10825433 TI - Effect of temperature on the formation and inactivation of syringomycin E pores in human red blood cells and bimolecular lipid membranes. AB - The effects of temperature on the formation and inactivation of syringomycin E (SRE) pores were investigated with human red blood cells (RBCs) and lipid bilayer membranes (BLMs). SRE enhanced the RBC membrane permeability of 86Rb and monomeric hemoglobin in a temperature dependent manner. The kinetics of 86Rb and hemoglobin effluxes were measured at different temperatures and pore formation was found to be only slightly affected, while inactivation was strongly influenced by temperature. At 37 degrees C, SRE pore inactivation began 15 min after and at 20 degrees C, 40 min after SRE addition. At 6 degrees C, below the phase transition temperature of the major lipid components of the RBC membrane, no inactivation occurred for as long as 90 min. With BLMs, SRE induced a large current that remained stable at 14 degrees C, but at 23 degrees C it decreased over time while the single channel conductance and dwell time did not change. The results show that the temperature dependent inactivation of SRE pores is due to a decrease in the number of open pores. PMID- 10825434 TI - Molecular dynamics study on lipid A from Escherichia coli: insights into its mechanism of biological action. AB - Structural properties of the Escherichia coli lipid A moiety were analysed by means of molecular mechanics and molecular dynamics simulations and compared to synthetic monophospho and dephospho analogues with different biological activities in the Limulus assay. The conformation of glucosamine disaccharide headgroup, order and packing of fatty acid chains, solvation of phosphate groups, coordination by water molecules, sodium counterions and models of cationic amino acid side chains were described in terms of mean values, mean residence times, radial distribution functions, coordination numbers, solvation and interaction energies. Solvation and polar interactions of the phosphate groups were correlated to known biological activities the lipid A variants. The observed relationship between the biological effect and the number and position of the phosphate groups were explained with the help of simple mechanistic models of lipid A action. The possible mechanism of action involving specific binding of lipid A disaccharide headgroup to cationic residues of a receptor model was compared with an alternative mechanism, which assumes a relationship between the ability to adopt non-lamellar supramolecular structures and the biological activity. Conclusions are drawn about the probable mode of lipid A action. Implications for rational drug design of endotoxin-neutralising agents are discussed. PMID- 10825435 TI - Modulation of volume regulated anion current by I(Cln). AB - I(Cln), a cytosolic protein associated with a nucleotide-sensitive chloride current, may be involved in the regulation of a volume-regulated anion current (VRAC) associated with hypotonic cell swelling. We have determined the nucleic acid sequences of I(Cln) from human tsA201a, colonic (T84) and myeloma (RPMI 8826) cell lines. The amino acid sequences are highly homologous (>/=99%) to each other but less homologous to I(Cln) protein from other species. Using the whole cell patch clamp technique, we examined the effect of I(Cln) protein expression levels on VRAC properties during a hyposmotic challenge. Overexpression of T84 or RPMI 8226-derived I(Cln) protein in tsA201a cells results in a more than 9-fold increase in the rate of VRAC activation over control values, while having no effect on VRAC inactivation properties. Underexpression of endogenous I(Cln) protein in tsA201a cells using antisense oligonucleotides results in a more than 180-fold decrease in VRAC activation rate as compared to control values. These results indicate that I(Cln) protein expression modulates VRAC activation but not inactivation. PMID- 10825436 TI - Rabbit nasal mucosa: nanospheres coated with polypeptides bound to specific anti polypeptide IgG are better transported than nanospheres coated with polypeptides or IgG alone. AB - In rabbit nasal mucosa, polypeptides and polypeptide-coated nanospheres are actively transported from lumen to blood by M-cells present in specialized transport areas of the epithelium. The largest transport is shown here to occur when some molecules of the polypeptides coating the nanospheres, after adsorption, are bound to the specific anti-polypeptide IgG, e.g. when insulin is bound to the anti-insulin IgG. The transport kinetics of nanospheres coated by insulin bound to its antibody, as a function of bead concentration or of the antibody/insulin coating ratio, have been analyzed. On this basis it was possible to assess the maximal transport capacity of the epithelium and to calculate the percentage of M-cells involved. PMID- 10825437 TI - Influence of band 3 protein absence and skeletal structures on amphiphile- and Ca(2+)-induced shape alterations in erythrocytes: a study with lamprey (Lampetra fluviatilis), trout (Onchorhynchus mykiss) and human erythrocytes. AB - Amphiphiles which induce either spiculated (echinocytic) or invaginated (stomatocytic) shapes in human erythrocytes, and ionophore A23187 plus Ca(2+), were studied for their capacity to induce shape alterations, vesiculation and hemolysis in the morphologically and structurally different lamprey and trout erythrocytes. Both qualitative and quantitative differences were found. Amphiphiles induced no gross morphological changes in the non-axisymmetric stomatocyte-like lamprey erythrocyte or in the flat ellipsoidal trout erythrocyte, besides a rounding up at higher amphiphile concentrations. No shapes with large broad spicula were seen. Nevertheless, some of the 'echinocytogenic' amphiphiles induced plasma membrane protrusions in lamprey and trout erythrocytes, from where exovesicles were shed. In trout erythrocytes, occurrence of corrugations at the cell rim preceded protrusion formation. Other 'echinocytogenic' amphiphiles induced invaginations in lamprey erythrocytes. The 'stomatocytogenic' amphiphiles induced invaginations in both lamprey and trout erythrocytes. Surprisingly, in trout erythrocytes, some protrusions also occurred. Some of the amphiphiles hemolyzed lamprey, trout and human erythrocytes at a significantly different concentration/membrane area. Ionophore A23187 plus Ca(2+) induced membrane protrusions and sphering in human and trout erythrocytes; however, the lamprey erythrocyte remained unperturbed. The shape alterations in lamprey erythrocytes, we suggest, are characterized by weak membrane skeleton lipid bilayer interactions, due to band 3 protein and ankyrin deficiency. In trout erythrocyte, the marginal band of microtubules appears to strongly influence cell shape. Furthermore, the presence of intermediate filaments and nuclei, additionally affecting the cell membrane shear elasticity, apparently influences cell shape changes in lamprey and trout erythrocytes. The different types of shape alterations induced by certain amphiphiles in the cell types indicates that their plasma membrane phospholipid composition differs. PMID- 10825438 TI - Functional studies of a chimeric protein containing portions of the Na(+)/glucose and Na(+)/myo-inositol cotransporters. AB - We obtained cDNA chimeras between Na/glucose cotransporter (SGLT1) and the homologous Na(+)/myo-inositol cotransporter (SMIT) by creating random chimeras in plasmids. Of 12 chimeras, two were functional when expressed in Xenopus laevis oocytes but, upon sequencing, only one of them (C1) produced an actual chimeric protein. In C1, the first 69 amino acids of SGLT1 were replaced by the corresponding 50 amino acids of SMIT. C1 transports the same sugars as does SGLT1. C1's affinity for all sugar substrates was systematically increased by a factor of 3.3+/-0.4 but the V(max) was diminished by a factor of 15-40. In contrast, the cotransport affinity for Na(+) was unchanged. The surface expression of C1 was one seventh that of SGLT1, which explains part of the reduced V(max) and implies a significant reduction in turnover rate. N-terminal truncated constructs of SGLT1 cDNA showed that deleting amino acids 2-14 does not affect cotransporter activity, but that the pentapeptide T(14)RPVET(19) is important for normal levels of SGLT1 current. The main result of a kinetic analysis of the systematic increase in apparent affinity for sugars, together with the intact Na apparent affinity, suggests enhanced access to the sugar binding site in C1. PMID- 10825439 TI - Comparative analysis of the isoform expression pattern of Ca(2+)-regulatory membrane proteins in fast-twitch, slow-twitch, cardiac, neonatal and chronic low frequency stimulated muscle fibers. AB - Although all muscle cells generate contractile forces by means of organized filament systems, isoform expression patterns of contractile and regulatory proteins in heart are not identical compared to developing, conditioned or mature skeletal muscles. In order to determine biochemical parameters that may reflect functional variations in the Ca(2+)-regulatory membrane systems of different muscle types, we performed a comparative immunoblot analysis of key membrane proteins involved in ion homeostasis. Cardiac isoforms of the alpha(1) dihydropyridine receptor, Ca(2+)-ATPase and calsequestrin are also present in skeletal muscle and are up-regulated in chronic low-frequency stimulated fast muscle. In contrast, the cardiac RyR2 isoform of the Ca(2+)-release channel was not found in slow muscle but was detectable in neonatal skeletal muscle. Up regulation of RyR2 in conditioned muscle was probably due to degeneration regeneration processes. Fiber type-specific differences were also detected in the abundance of auxiliary subunits of the dihydropyridine receptor, the ryanodine receptor and the Ca(2+)-ATPase, as well as triad markers and various Ca(2+) binding and ion-regulatory proteins. Hence, the variation in innervation of different types of muscle appears to have a profound influence on the levels and pattern of isoform expression of Ca(2+)-regulatory membrane proteins reflecting differences in the regulation of Ca(2+)-homeostasis. However, independent of the muscle cell type, key Ca(2+)-regulatory proteins exist as oligomeric complexes under native conditions. PMID- 10825440 TI - Palmitoylation of a pulmonary surfactant protein C analogue affects the surface associated lipid reservoir and film stability. AB - Surfactant protein C (SP-C) is a lipopeptide that contains two thioester-linked palmitoyl groups and is considered to be important for formation of the alveolar surface active lipid film. Here, a non- or dipalmitoylated SP-C analogue (SP C(Leu)), in which all helical Val residues were replaced with Leu and Cys-5 and Cys-6 were replaced with Ser, was tested for surface activity in a captive bubble system (CBS). SP-C(Leu), either palmitoylated at Ser-5 and Ser-6 or non palmitoylated, was added to mixtures of 1, 2-dipalmitoyl-sn-glycero-3 phosphocholine (DPPC)/phosphatidyl glycerol (PG)/palmitic acid (PA), 68:22:9, (by mass) at a concentration of 2 and 5%. With 2% peptide, surface film formation was rapid, reaching a surface tension below 25 mN/m within 5 s, but the samples with 5% SP-C(Leu) required more than 20 s to reach values below 25 mN/m. Minimum surface tension for the samples with dipalmitoylated SP-C(Leu) was below 1.5 mN/m and very stable, as the surface tension increased by less than 0.5 mN/m within 10 min at constant bubble volume. Minimum surface tension for the non-palmitoylated SP-C(Leu) was approximately 2 and 5 mN/m for 2 and 5% peptide, respectively, but the films were less stable as seen by frequent bubble clicking at low surface tensions. Films with dipalmitoylated SP-C(Leu) that were dynamically cycled at 20 30 cycles/min were substantially less compressible at a surface tension of 20 mN/m (0.007 m/mN) than those that contained the non-palmitoylated peptide (0.02 m/mN). After subphase depletion, the incorporation of lipids into the surface active film during initial bubble expansion occurred at a relatively low surface tension (about 35 mN/m) for the samples with dipalmitoylated SP-C(Leu) compared to approximately 45 mN/m for those containing the non-palmitoylated peptide. Furthermore, for samples that contained non-palmitoylated SP-C(Leu), the ability to reach near zero stable surface tension was lost after a few adsorption steps, whereas with the dipalmitoylated peptide the film quality did not deteriorate even after more than 10 expansion steps and the incorporation of reservoir material equivalent to more than two monolayers. It appears that the covalently linked palmitoyl groups of the SP-C analogue studied are important for the mechanical stability of the lipid film, for the capacity to incorporate material from the reservoir into the surface active film upon area expansion, and for the low film compressibility of dynamically cycled films. PMID- 10825441 TI - Pulmonary surfactant protein B: a structural model and a functional analogue. AB - Surfactant proteins B and C (SP-B and SP-C), together with phospholipids, are important constituents of pulmonary surfactant and of preparations used for treatment of respiratory distress syndrome (RDS). SP-B belongs to the saposin family of homologous proteins, which include other lipid-interacting proteins, like the membranolytic NK-lysin. SP-B, in contrast to other saposins, is hydrophobic and a disulfide-linked dimer, and its mechanism of action is not known. A model of the three-dimensional structure of one SP-B subunit was generated from the structure of monomeric NK-lysin determined by nuclear magnetic resonance, and the SP-B dimer was formed by joining two subunits via the intersubunit disulfide bond Cys48-Cys48'. After energy minimization, intersubunit hydrogen bonds/ion pairs were formed between the strictly conserved residues Glu51 and Arg52, which creates a central non-polar region located in between two clusters of positively charged residues. The structural features support a function of SP-B in cross-linking of lipid membranes. Mixtures of phospholipids, an SP-C analogue and polymyxin B (which cross-links lipid vesicles but is structurally unrelated to SP-B) exhibit in vitro surface activity which is indistinguishable from that of analogous mixtures containing SP-B instead of polymyxin B. This suggests an avenue for identification of SP-B analogues that can be used in synthetic surfactants for treatment of RDS. PMID- 10825442 TI - A Ca(2+)- and voltage-modulated flagellar ion channel is a component of the mechanoshock response in the unicellular green alga Spermatozopsis similis. AB - In flagellate green algae, behavioral responses to photo- and mechanoshock are induced by different external stimuli within 10-15 ms. In the accompanying changes in flagella beat, Ca(2+) has important regulatory roles. Although the axonemal Ca(2+) responsive elements are well characterized, analyses of flagellar channels involved in Ca(2+) signalling as well as other ion channels at the single-channel level were not yet conducted in green algae. To gain a further understanding of these important signaling elements in movement responses, intact flagella of Spermatozopsis similis were isolated and characterized and the solubilized flagellar membrane proteins were reconstituted into liposomes. We observed three types of channel activity, two of which were weakly anion and cation-selective and in the high-conductance regime typical for porin-like solute channels. The dominating channel activity was a voltage dependent, rectifying, low conductance (Lambda=80 pS in 50 mM KCl) cation-selective channel modulated by, and highly permeable to, Ca(2+) ions (SFC1: Spermatozopsis flagellar cation channel 1). Depolarizations necessary to activate SFC1 probably only occur in vivo during avoidance reactions of this alga. Ca(2+)-activation of SFC1 points to a direct link to Ca(2+)-mediated signaling pathway(s) in the flagella. Both the response to mechanoshock and SFC1 activity were inhibited by Gd(3+) and Ba(2+), thus supporting our assumption that SFC1 represents a major flagellar ion channel involved in this green algal avoidance reaction. PMID- 10825443 TI - Selective targeting of immunoliposomal doxorubicin against human multiple myeloma in vitro and ex vivo. AB - Circulating malignant CD19(+) B cells have been implicated in the pathogenesis and relapse of multiple myeloma (MM). This study investigated the therapeutic applicability of using long-circulating liposome-encapsulated doxorubicin (DXR) targeted against the internalizing CD19 antigens present on human MM cells. In vitro binding studies using the CD19(+) MM cell line ARH77 demonstrated that CD19 directed immunoliposomes (SIL[anti-CD19]) specifically attached to these cells. Formulations of immunoliposomal doxorubicin (DXR-SIL[anti-CD19]) showed a higher association with, and higher cytotoxicity against, ARH77 cells than did non targeted liposomal doxorubicin (DXR-SL) or isotype-matched controls (DXR NSIL[IgG2a]). By using the pH-sensitive fluorophore, 1-hydroxypyrene-3,6, 8 trisulfonic acid, binding of SIL[anti-CD19] to CD19 antigens was shown to trigger receptor-mediated internalization of the antibody-antigen complexes into endosomes. Targeting of SIL[anti-CD19] to CD19(+) B cells was also demonstrated in a heterogeneous mixture of peripheral blood mononuclear cells (PBMC) from MM patients. A decrease in cellular DNA (which is an indicator of apoptosis) caused by the cytotoxicity of DXR-SIL[anti-CD19] to myeloma PBMC was determined by using flow cytometry. While PBMC treatment with free DXR resulted in non-specific cytotoxicity to both B and T cells, DXR-SL were only minimally cytotoxic to either. In contrast, DXR-SIL[anti-CD19] were selectively cytotoxic for B cells in PBMC, indicating that this treatment may be effective in eliminating circulating malignant B cells in MM patients. PMID- 10825444 TI - Theoretical calculations of the permeability of monensin-cation complexes in model bio-membranes. AB - Monensin is one of the best-characterized ionophores; it functions in the electroneutral exchange of cations between the extracellular and cytoplasmic sides of cell membranes. The X-ray crystal structures of monensin in free acid form and in complex with Na(+), K(+) and Ag(+) are known and we have recently measured the diffusion rates of monensin in free acid form (Mo-H) and in complex with Na(+) (Mo-Na) and with K(+) (Mo-K) using laser pulse techniques. The results have shown that Mo-H diffuses across the membrane one order of magnitude faster than Mo-Na and two orders of magnitude faster than Mo-K. Here, we report calculations of the translocation free energy of these complexes across the membrane along the most favorable path, i.e. the lowest free energy path. The calculations show that the most favorable orientation of monensin is with its hydrophobic furanyl and pyranyl moieties in the hydrocarbon region of the membrane and the carboxyl group and the cation at the water-membrane interface. Further, the calculations show that Mo-H is likely to be inserted deeper than Mo Na into the bilayer, and that the free energy barrier for transfer of Mo-H across the membrane is approximately 1 kcal/mol lower than for Mo-Na, in good agreement with our measurements. Our results show that the Mo-K complex is unlikely to diffuse across lipid bilayers in its X-ray crystal structure, in contrast to the Mo-H and Mo-Na complexes. Apparently, when diffusing across the membrane, the Mo K complex assumes a different conformation and/or thinning defects in the bilayer lower significantly the free energy barrier for the process. The suitability of the model for treating the membrane association of small molecules is discussed in view of the successes and failures observed for the monensin system. PMID- 10825446 TI - A Fourier transform infrared spectroscopic study of the interaction of alkaline earth cations with the negatively charged phospholipid 1, 2-dimyristoyl-sn glycero-3-phosphoglycerol. AB - The interaction of aqueous phospholipid dispersions of negatively charged 1,2 dimyristoyl-sn-glycero-3-phosphoglycerol, sodium salt (DMPG) with the divalent cations Mg(2+), Ca(2+) and Sr(2+) at equimolar ratios in 100 mM NaCl at pH 7 was investigated by Fourier transform infrared spectroscopy. The binding of the three cations induces a crystalline-like gel phase with highly ordered and rigid all trans acyl chains. These features are observed after storage below room temperature for 24 h. When the gel phase is heated after prolonged incubation at low temperature phase transitions into the liquid crystalline phase are observed at 58 degrees C for the DMPG:Sr(2+), 65 degrees C for the DMPG:Mg(2+), and 80 degrees C for the DMPG:Ca(2+) complex. By subsequent cooling from temperatures above T(m) these complexes retain the features of a liquid crystalline phase with disordered acyl chains until a metastable gel phase is formed at temperatures between 38 and 32 degrees C. This phase is characterized by predominantly all trans acyl chains, arranged in a loosely packed hexagonal or distorted hexagonal subcell lattice. Reheating the DMPG:Sr(2+) samples after a storage time of 2 h at 4 degrees C results in the transition of the metastable gel to the liquid crystalline phase at 35 degrees C. This phase transition into the liquid crystalline state at 35 degrees C is also observed for the Mg(2+) complex. However, for DMPG:Mg(2+) at higher temperatures, a partial recrystallization of the acyl chains occurs and the high temperature phase transition at 65 degrees C is also detected. In contrast, DMPG:Ca(2+) exhibits only the phase transition at 80 degrees C from the crystalline gel into the fluid state upon reheating. Below 20 degrees C, the rate of conversion from the metastable gel to a thermodynamically stable, crystalline-like gel phase decreases in the order Ca(2+)&z. Gt;Mg(2+)>Sr(2+). This conversion into the crystalline gel phase is accompanied by a complete dehydration of the phosphate groups in DMPG:Mg(2+) and by a reorientation of the polar lipid head groups in DMPG:Ca(2+) and in DMPG:Sr(2+). The primary binding sites of the cations are the PO(2)(-) groups of the phosphodiester moiety. Our infrared spectroscopic results suggest a deep penetration of the divalent cations into the polar head group region of DMPG bilayers, whereby the ester carbonyl groups, located in the interfacial region of the bilayers, are indirectly affected by strong hydrogen bonding of immobilized water molecules. In the liquid crystalline phase, the interaction of all three cations with DMPG is weak, but still observable in the infrared spectra of the DMPG:Ca(2+) complex by a slight ordering effect induced in the acyl chains, when compared to pure DMPG liposomes. PMID- 10825445 TI - Inhibitory effects of some purinergic agents on ecto-ATPase activity and pattern of stepwise ATP hydrolysis in rat liver plasma membranes. AB - Inhibitory effects of various purinergic compounds on the Mg(2+)-dependent enzymatic hydrolysis of [(3)H]ATP in rat liver plasma membranes were evaluated. Rat liver enzyme ecto-ATPase has a broad nucleotide-hydrolyzing activity, displays Michaelis-Menten kinetics with K(m) for ATP of 368+/-56 microM and is not sensitive to classical inhibitors of the ion-exchange and intracellular ATPases. P2-antagonists and diadenosine tetraphosphate (Ap(4)A) progressively and non-competitively inhibited ecto-ATPase activity with the following rank order of inhibitory potency: suramin (pIC(50), 4.570)>Reactive blue 2 (4.297)&z.Gt;Ap(4)A (3. 268)>pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid (PPADS) (2. 930). Slowly hydrolyzable P2 agonists ATPgammaS, ADPbetaS, alpha, beta-methylene ATP and beta,gamma-methylene ATP as well as the diadenosine polyphosphates Ap(3)A and Ap(5)A did not exert any inhibitory effects on the enzyme activity at concentration ranges of 10(-4)-10(-3) M. Thin-layer chromatography analysis of the formation of [(3)H]ATP metabolites indicated the presence of other enzyme activities on liver surface (ecto-ADPase and 5'-nucleotidase), participating in concert with ecto-ATPase in the nucleotide hydrolysis through the stepwise reactions ATP-->ADP-->AMP-->adenosine. A similar pattern of sequential [(3)H]ATP dephosphorylation still occurs in the presence of ecto-ATPase inhibitors suramin, Ap(4)A and PPADS, but the appearance of the ultimate reaction product, adenosine, was significantly delayed. In contrast, hydrolysis of [(3)H]ATP in the presence of Reactive blue 2 only followed the pattern ATP-->ADP, with formation of the subsequent metabolites AMP and adenosine being virtually eliminated. These data suggest that although nucleotide-binding sites of ecto-ATPase are distinct from those of P2 receptors, some purinergic agonists and antagonists can potentiate cellular responses to extracellular ATP through non-specific inhibition of the ensuing pathways of purine catabolism. PMID- 10825447 TI - Newly isolated archaerhodopsin from a strain of Chinese halobacteria and its proton pumping behavior. AB - A strain of extremely salt-loving halobacteria Halobacterium species xz515 from a salt lake in Tibet was isolated. SDS-polyacrylamide gel electrophoresis shows that there is only one protein on claret membrane, which is the same membrane fraction as purple membrane from Halobacterium salinarum, with a molecular weight close to bacteriorhodopsin (br). The purified retinal containing protein from xz515 has an absorption peak at around 550 nm. These facts indicate that it is a br-like protein. The partial sequence determination [H. Wang et al., Chin. Sci. Bull., 45 (2000)] shows that this br-like protein belongs to the archaerhodopsin family. The measurements of light-induced medium pH change in intact cells and cell envelope vesicles of xz515 suggest that this type of archaerhodopsin has a proton pumping function. However, the study about the dynamics of pumped protons across the membrane reveal that the proton release and proton uptake is in reverse order compared to br. The probable reason, attributing to regulating the rate of proton release is discussed. PMID- 10825448 TI - Diethyl pyrocarbonate inactivates CD39/ecto-ATPDase by modifying His-59. AB - Diethyl pyrocarbonate (DEPC) in conditions that favour carbethoxylation of histidyl residues strongly inactivated E-type ATPase activity of a rat lung membrane preparation, as well as ecto-ATPase activity of rat vessels and human Epstein-Barr virus-transformed B lymphocytes. Inactivation of the enzyme (up to 70%) achieved at concentrations of DEPC below 0.5 mM could be fully reversed by 200 mM hydroxylamine at pH 7.5, thus confirming histidine-selective modification. UTP effectively protected the enzyme activity from DEPC inactivation. This was taken to indicate that the conformation adopted by the enzyme molecule upon substrate binding was not compatible with DEPC reaching and/or modifying the relevant histidyl residue. Substrate activation curves were interpreted to show the enzyme molecule to be inactive, at all substrate concentrations tested, when the target histidyl residue had been modified by DEPC. Comparison of known sequences of CD39-like ecto-ATP(D)ases with the results on inactivation by DEPC revealed His-59 and His-251 (according to the human CD39 sequence) as equally possible targets of the inactivating DEPC modification. Potato apyrase lacks a homologue for the former residue, while the latter is preserved in the enzyme sequence. Therefore, this enzyme was exposed to DEPC, and since hydrolysis of ATP and ADP by potato apyrase was insensitive to modification with DEPC, it was concluded that His-59 is the essential residue in CD39 that is affected by DEPC modification in a way that causes inactivation of the enzyme. PMID- 10825449 TI - Hybrid polar compounds produce a positive shift in the surface dipole potential of self-assembled phospholipid monolayers. AB - Hybrid polar compounds (HPCs) are powerful inducers of terminal differentiation of various types of tumors, including Friend murine erythroleukemia cells (MELCs). They are known to act synergistically with an increase in the extracellular concentration of cations, which causes a positive shift in the negative value of the ionic surface potential. Two HPCs, hexamethylenebisacetamide (HMBA) and suberoylanilide hydroxamic acid (SAHA), were adsorbed on self-assembled phospholipid monolayers supported on a mercury drop and the shift in the surface dipole potential chi of the lipid film due to their adsorption was estimated from charge measurements. At their optimal concentrations for inducing MELC terminal differentiation (5 mM for HMBA and 2.6 microM for SAHA), these HPCs cause a chi shift of about 15-20 mV, positive toward the hydrocarbon tails, both on neutral phosphatidylcholine films and on negatively or positively charged phosphatidylserine films. This strongly suggests that the nonspecific effect of HPCs of different structure in inducing cancer cells to rescue their differentiation program is related to a positive chi shift on the extracellular side of the cell membrane. PMID- 10825450 TI - Characterisation of three novel cationic lipids as liposomal complexes with DNA. AB - Cationic lipids (CLs) are being increasingly exploited as transfection vectors for the delivery of DNA into eukaryotic cells. To obtain further insight to the complex formation and interactions between cationic liposomes and DNA, we characterised three novel cationic lipids, viz. bis[2-(11 phenoxyundecanoate)ethyl]-dimethylammonium bromide, N-hexadecyl-N-?10-[O-(4 acetoxy)-phenylundecanoate]ethyl?- dimethylammonium bromide, and bis[2-(11 butyloxyundecanoate)ethyl]dimethylammonium bromide. These lipids bear the same charged headgroup yet have different hydrophobic parts. Accordingly, we may anticipate their electrostatic interactions with DNA to be similar while differing in both thermal phase behaviour and physicochemical properties of their complexes with DNA. In keeping with the above all three lipids formed complexes with DNA as evidenced by light scattering, fluorescence spectroscopy and Langmuir film balance. Differential scanning calorimetry revealed very different phase behaviours for the binary mixtures of the three CLs with dimyristoylphosphatidylcholine and also provided evidence for DNA-induced lipid phase separation. These data were confirmed by compression isotherms and fluorescence microscopy of monolayers residing on an aqueous buffer, recorded both in the presence and absence of DNA. Importantly, binding to cationic liposomes appears to prevent thermal denaturation of DNA upon heating of the complexes. Likewise, renaturation of heat-treated DNA complexed with the cationic liposomes appears to be abolished as well. PMID- 10825451 TI - The abnormality of glucose transporter in the erythrocyte membrane of Chinese type 2 diabetic patients. AB - Type 2 diabetes mellitus is characterized by impaired glucose uptake. With a photometric method of recording the erythrocyte suspension absorption during the course of glucose transport across the membranes, we observed that the initial rate of glucose zero-trans entry was decreased significantly in 30 Chinese type 2 diabetic patients as compared to 25 healthy controls. The rate of glucose infinite-cis efflux exhibited no difference between the patients and controls. The measurement of temperature dependence of glucose transport showed that the activation energy for glucose entry was increased in diabetic patients. The inhibitory constant of glucose entry by cytochalasin B (CB) in patients was similar to that of the controls. However, we found that the inhibitory constant was increased significantly in the patient erythrocytes after phloretin treatment. After the erythrocytes were made into stripped white ghosts, the fluorescence quenching experiment was performed. Glucose, CB and phloretin can quench the fluorescence of tryptophan residues in the glucose transporter 1, GLUT1. The abnormality of fluorescence quenching in the erythrocyte membranes of patients was observed. The transfer tendency of tryptophan residues from the hydrophilic environment to the hydrophobic environment was decreased in patient ghosts as binding with glucose, and the opposite tendency appeared as CB and phloretin instead of glucose. We conclude that the decreased in glucose entry in the erythrocyte membranes of diabetic patients was due to the GLUT1 change in structure - mostly the outer domain of the glucose transporter. PMID- 10825452 TI - Structural and functional characteristics and tissue distribution pattern of rat OCTN1, an organic cation transporter, cloned from placenta. AB - This report describes the structure, function, and tissue distribution pattern of rat OCTN1 (novel organic cation transporter 1). The rat OCTN1 cDNA was isolated from a rat placental cDNA library. The cDNA is 2258 bp long and codes for a protein of 553 amino acids. Its amino acid sequence bears high homology to human OCTN1 (85% identity) and rat OCTN2 (74% identity). When expressed heterologously in mammalian cells, rat OCTN1 mediates Na(+)-independent and pH-dependent transport of the prototypical organic cation tetraethylammonium. The transporter interacts with a variety of structurally diverse organic cations such as desipramine, dimethylamiloride, cimetidine, procainamide, and verapamil. Carnitine, a zwitterion, interacts with rat OCTN1 with a very low affinity. However, the transport of carnitine via rat OCTN1 is not evident in the presence or absence of Na(+). We conclude that rat OCTN1 is a multispecific organic cation transporter. OCTN1-specific mRNA transcripts are present in a wide variety of tissues in the rat, principally in the liver, intestine, kidney, brain, heart and placenta. In situ hybridization shows the distribution pattern of the transcripts in the brain (cerebellum, hippocampus and cortex), kidney (cortex and medulla with relatively more abundance in the cortical-medullary junction), heart (myocardium and valves) and placenta (labyrinthine zone). PMID- 10825453 TI - Membrane topology of the Na(+)/citrate transporter CitS of Klebsiella pneumoniae by insertion mutagenesis. AB - The sodium ion dependent citrate transporter of Klebsiella pneumoniae (CitS) is a member of the bacterial 2-hydroxycarboxylate transporter family. Membrane topology models of the protein, largely based on reporter molecule fusions to C terminally truncated CitS molecules, indicate that the protein traverses the membrane 11 times with the NH(2)-terminus in the cytoplasm and the COOH-terminus in the periplasm. Furthermore, the structure is characterized by unusual long loops in the COOH-terminal half of the protein: one hydrophobic segment between transmembrane segments V and VI in the periplasm and three long loops connecting transmembrane segments VI and VII, VIII and IX and X and XI in the cytoplasm. The 10 kDa biotin acceptor domain and six consecutive His residues (His-tag) were inserted at different positions in the four long loops and the effect on transport activity and protein stability was analyzed. Six out of seven insertion mutants were stably expressed and three of these had retained significant transport activity. The sidedness of the tags in the mutants that tolerated the insertion was determined by proteolysis experiments. The results support the 11 transmembrane segment model that was based upon truncated CitS proteins. PMID- 10825454 TI - Dye removal, catalytic activity and 2D crystallization of chloroplast H(+)-ATP synthase purified by blue native electrophoresis. AB - The proton-ATP synthase of thylakoid membranes from spinach chloroplasts (CF(O)F(1)) and its subcomplexes CF(O) and CF(1) were isolated by blue native electrophoresis (BN-PAGE) [Neff, D. and Dencher, N.A. (1999) Biochem. Biophys. Res. Commun. 259, 569-575] and subsequently electroeluted from the gel. A method was developed to remove most of the dye Coomassie G-250 (CBG) using gel filtration, a prerequisite for many biophysical investigations. The dye was removed from the electroeluted CF(O)F(1), CF(O) or CF(1) and exchanged with the detergent CHAPS. ATP hydrolysis activity of CF(1) and ATP synthesis activity of reconstituted CF(O)F(1) were determined before and after dye removal. The secondary structure of CF(O) was studied by CD spectroscopy in the presence and the absence of the dye. CBG neither abolishes the catalytic activity of the isolated CF(O)F(1) and CF(1) nor affects the subunit composition and the high alpha-helical content of CF(O). In crystallization attempts, 2D arrays of CF(O)F(1) and of CF(O) before and after dye removal were obtained. In the aggregates of CF(O), circular structures with a mean diameter of 6.7 nm were observed. Our results indicate that the combination of BN-PAGE and dye removal by gel filtration is a suitable approach to obtain catalytically active protein complexes for further functional and structural characterization. PMID- 10825455 TI - Fatty acid binding sites of serum albumin probed by non-linear spin-label EPR. AB - A novel form of non-linear EPR spectroscopy, viz. the first harmonic absorption spectrum recorded in phase quadrature with respect to the Zeeman field modulation, is used here to investigate spin-lattice relaxation enhancements of nitroxide spin labels bound to serum albumin that are induced by spin-spin interactions with aqueous paramagnetic ions. The advantage of this EPR method is that it is directly sensitive to spin-lattice relaxation and affected relatively little by other spectral parameters (Livshits et al., J. Magn. Reson. 133 (1998) 79-91). Relaxation enhancements by ferricyanide of bound fatty acids (n-SASL) spin-labelled at different positions, n, in the chain are compared with those of different maleimide spin label derivatives attached at the single free -SH group, as well as with those of the spin labels free in solution. It was found that: (1) the encounter frequency of ferricyanide with 5-SASL and 12-SASL bound to serum albumin is more than two times less than that with 16-SASL; (2) the accessibility of ferricyanide to 16-SASL is comparable to that of the more immobilised covalently bound spin labels; and (3) the absolute values of the encounter frequencies for the bound spin-labelled fatty acids are approximately a factor of ten smaller than for the corresponding free spin labels, but the latter show a dependence on position of labelling that is similar to the bound labels. A kinetic scheme that is consistent with these relative differences involves rapid reversible transitions between an 'open' and 'closed' state, in which interaction with aqueous paramagnetic agents is possible only in the 'open' state. The equilibrium strongly favours the 'closed' state, which is further enhanced at low temperatures. PMID- 10825456 TI - Characterization of Na/Ca exchange in plasmalemmal vesicles from zona fasciculata cells of the bovine adrenal gland. AB - The presence of an Na/Ca exchange system in fasciculata cells of the bovine adrenal gland was tested using isolated plasmalemmal vesicles. In the presence of an outwardly Na(+) gradient, Ca(2+) uptake was about 2-fold higher than in K(+) condition. Li(+) did not substitute for Na(+) and 5 mM Ni(2+) inhibited Ca(2+) uptake. Ca(2+) efflux from Ca(2+)-loaded vesicles was Na(+)-stimulated and Ni(2+) inhibited. The saturable part of Na(+)-dependent Ca(2+) uptake displayed Michaelis-Menten kinetics. The relationship of Na(+)-dependent Ca(2+) uptake versus intravesicular Na(+) concentration was sigmoid (apparent K(0.5) approximately 24 mM; Hill number approximately 3) and Na(+) acted on V(max) without significant effect on K(m). Na(+)-stimulated Ca(2+) uptake was temperature-dependent (apparent Q(10) approximately 2.2). The inhibition properties of several divalent cations (Cd(2+), Sr(2+), Ni(2+), Ba(2+), Mn(2+), Mg(2+)) were tested and were similar to those observed in kidney basolateral membrane. The above results indicate the presence of an Na/Ca exchanger located on plasma membrane of zona fasciculata cells of bovine adrenal gland. This exchanger displays similarities with that of renal basolateral cell membrane. PMID- 10825457 TI - Characterization of an oligopeptide transporter in renal lysosomes. AB - Renal lysosomes play a major role in catabolism of plasma proteins. Final products of this catabolism include dipeptides and tripeptides that must be exported to the cytosol for hydrolysis. The aim of the present study was to determine whether an oligopeptide transporter is present in the renal lysosomal membrane that could mediate this export. The existence of an oligopeptide transporter was probed with the uptake of glycylglutamine (Gly-Gln) by membrane vesicles prepared from renal lysosomes. Kinetic analysis showed the presence of a single transporter with a K(m) of 8.77 mM for the uptake of Gly-Gln. The Gly-Gln uptake was energized by the imposition of an inwardly directed proton gradient (pH(out) 5.0/pH(in) 7.3) and membrane potential (outside positive/inside negative) resulting in overshoot. The Gly-Gln uptake was inhibited by the presence of dipeptides and tripeptides, but not amino acids. Western blot analysis of lysosomal membrane proteins with Pept-1 (an oligopeptide transporter) antibody as the probe showed the presence of an immunoreactive protein. This immunoreaction was abolished when the antiserum was preabsorbed with the Pept-1 epitope (0.5 microg/ml). In conclusion, the present data show the existence of a low-affinity dipeptide transporter in the renal lysosomal membrane that appears to belong to the Pept family of transporters. The function of this transporter appears to be to prevent accumulation of dipeptides in renal lysosomes. PMID- 10825458 TI - Characterization and partial purification of liver glucose transporter GLUT2. AB - GLUT2, the major facilitative glucose transporter isoform expressed in hepatocytes, pancreatic beta-cells, and absorptive epithelial cells, is unique not only with its low affinity and broad substrate specificity as a glucose transporter, but also with its implied function as a glucose-sensor. As a first essential step toward structural and biochemical elucidation of these unique, GLUT2 functions, we describe here the differential solubilization and DEAE-column chromatography of rat hepatocyte GLUT2 protein and its reconstitution into liposomes. The reconstituted GLUT2 bound cytochalasin B in a saturable manner with an apparent dissociation constant (K(d)) of 2.3 x 10(-6) M and a total binding capacity (B(T)) of 8.1 nmol per mg protein. The binding was completely abolished by 2% mercury chloride, but not affected by cytochalasin E. Significantly, the binding was also not affected by 500 mM D-glucose or 3-O methyl D-glucose (3OMG). The purified GLUT2 catalyzed mercury chloride-sensitive 3OMG uptake, and cytochalasin B inhibited this 3OMG uptake. The inhibition was dose-dependent with respect to cytochalasin B, but was independent of 3OMG concentrations. These findings demonstrate that our solubilized GLUT2 reconstituted in liposomes is at least 60% pure and functional, and that GLUT2 is indeed unique in that its cytochalasin B binding is not affected by its substrate (D-glucose) binding. Our partially purified GLUT2 reconstituted in vesicles will be useful in biochemical and structural elucidation of GLUT2 as a glucose transporter and as a possible glucose sensor. PMID- 10825459 TI - Use of echinacea in medicine. AB - Echinacea, also known as the purple coneflower, is an herbal medicine that has been used for centuries, customarily as a treatment for the common cold, coughs, bronchitis, upper respiratory infections, and some inflammatory conditions. Research on echinacea, including clinical trials, is limited and largely in German. More information is needed before a definitive statement about the efficacy of echinacea can be made. Future work needs to clearly identify the species of echinacea and distinguish between the efficacy of the different plant parts (roots versus upper plant parts). Although many of the active compounds of echinacea have been identified, the mechanism of action is not known, nor is the bioavailability, relative potency, or synergistic effects of the active compounds known. Interpretation of existing literature suggests that echinacea should be used as a treatment for illness, not as a means for prevention of illness. The consensus of the studies reviewed in this article is that echinacea is indeed effective in reducing the duration and severity of symptoms, but that this effect is noted only with certain preparations of echinacea. Studies show that the plant and its active components affect the phagocytic immune system, but not the specifically acquired immune system. PMID- 10825460 TI - Rapid desensitization and slow recovery of the cyclic AMP response mediated by histamine H(2) receptors in the U937 cell line. AB - The present study focused on the desensitization process of the H(2) receptor in U937 cells and the recovery of the cyclic AMP (cAMP) response. Treatment of U937 leukemic cells with the H(2) histamine receptor agonists (+/-)-N(1)-[3-(3, 4 difluorophenyl)-3-(pyridin-2-yl)propyl]-N(2)-[3-(1H-imidazol-4-yl)p ropyl]guanidine (BU-E-75) and amthamine produced a rapid desensitization characterized by decreased cAMP production (T(1/2) = 20 min). Pretreatment with 10 microM BU-E-75 did not induce modifications in the responses to prostaglandin E(2), isoproterenol, or forskolin. H(2) receptor desensitization was not affected by protein kinase A and C inhibitors, but was reduced drastically by Zn(2+) and heparin, known to act as inhibitors of G protein-coupled receptor kinases. Recovery studies of the cAMP response showed that cAMP levels reached 50% of the initial values within 5 hr. Furthermore, desensitization produced an important decrease in the basal level of this cyclic nucleotide. The minimal value was observed 12 hr later, and corresponded to approximately 1.3% of the initial basal level (7.5 vs 0.1 pmol/10(6) cells). This result could be explained by an increase in phosphodiesterase activity following 10 microM BU-E-75 treatment. When cells were exposed for 2 hr to an H(2) agonist, binding assays showed no modification in the number of H(2) receptors; internalization began just after 8 hr. Although the initial desensitization seems to involve G protein-coupled receptor kinases, results indicate that additional mechanisms of regulation were triggered by the H(2) agonists. PMID- 10825461 TI - Ligand-independent activation of estrogen receptor function by 3, 3' diindolylmethane in human breast cancer cells. AB - 3,3'-Diindolylmethane (DIM), a major in vivo product of acid-catalyzed oligomerization of indole-3-carbinol (I3C), is a promising anticancer agent present in vegetables of the Brassica genus. We investigated the effects of DIM on estrogen-regulated events in human breast cancer cells and found that DIM was a promoter-specific activator of estrogen receptor (ER) function in the absence of 17beta-estradiol (E(2)). DIM weakly inhibited the E(2)-induced proliferation of ER-containing MCF-7 cells and induced proliferation of these cells in the absence of steroid, by approximately 60% of the E(2) response. DIM had little effect on proliferation of ER-deficient MDA-MB-231 cells, suggesting that it is not generally toxic at these concentrations. Although DIM did not bind to the ER in this concentration range, as shown by a competitive ER binding assay, it activated the ER to a DNA-binding species. DIM increased the level of transcripts for the endogenous pS2 gene and activated the estrogen-responsive pERE-vit-CAT and pS2-tk-CAT reporter plasmids in transiently transfected MCF-7 cells. In contrast, DIM failed to activate transcription of the simple E(2)- and diethylstilbesterol-responsive reporter construct pATC2. The estrogen antagonist ICI 182780 (7alpha-[9-[(4,4,5,5, 5-pentafluoropentyl)sulfonyl]nonyl]-estra 1,3,5(10)-triene-3, 17beta-diol) was effective against DIM-induced transcriptional activity of the pERE-vit-CAT reporter, which further supports the hypothesis that DIM is acting through the ER. We demonstrated that ligand independent activation of the ER in MCF-7 cells could be produced following treatment with the D1 dopamine receptor agonist SKF-82958 [(+/-)6-chloro-7,8 dihydroxy-3-allyl-1-phenyl-2,3,4, 5-tetrahydro-1H-3-benzazepinehydrobromide]. We also demonstrated that the agonist effects of SKF-82958 and DIM, but not of E(2), could be blocked by co-treatment with the protein kinase A (PKA) inhibitor H-89 (N-[2-(p-bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide). These results have uncovered a promoter-specific, ligand-independent activation of ER signaling for DIM that may require activation by PKA, and suggest that this major I3C product may be a selective activator of ER function. PMID- 10825462 TI - Deficient homologous desensitization of formyl peptide receptors stably expressed in undifferentiated HL-60 cells. AB - The ability of formyl peptide receptors (FPRs) stably expressed in undifferentiated HL-60 cells to undergo ligand-induced desensitization was compared with their ability in normal and vector-transfected HL-60 cells following granulocyte differentiation with DMSO. fMet-Leu-Phe failed to induce uncoupling of FPRs from G-proteins in FPR-transfected cells, whereas uncoupling was induced in differentiated HL-60 cells and differentiated vector-transfected HL-60 cells, as determined by ligand-stimulated guanosine 5'-(gamma thio)triphosphate (GTPgammaS) binding and GTPgammaS inhibition of fMet-Leu-Phe binding to isolated membranes. Immunoprecipitation of Galpha(i2) from solubilized, azidoanalide (AA-gammaGTP) photolabeled membranes showed that receptors in desensitized FPR-transfected HL-60 cells remained coupled to Galpha(i2), whereas desensitized receptors in differentiated HL-60 cell membranes were uncoupled from Galpha(i2). As determined by immunoblotting, Galpha(i2) expression was similar in undifferentiated and differentiated HL-60 cells and FPR transfected cells. Ligand-stimulated receptor internalization and desensitization of calcium redistribution were similar in all three groups of cells. Immunoblotting also indicated that G-protein-coupled receptor kinases (GRKs) 2 and 4 were present in undifferentiated FPR-transfected HL-60 cells at 50% of the level seen in differentiated HL-60 cells. However, differentiation did not increase GRK2 or GRK4 expression, indicating that differences in GRK expression do not explain deficient desensitization. The data indicated that undifferentiated HL-60 cells are unable to induce homologous desensitization of FPRs. PMID- 10825463 TI - Effect of 3'-methoxy-4'-nitroflavone on benzo[a]pyrene toxicity. Aryl hydrocarbon receptor-dependent and -independent mechanisms. AB - This laboratory has studied a number of flavone derivatives for aryl hydrocarbon receptor (AhR) agonist and antagonist potential using cell-free and cell culture systems. The current report extends these investigations by testing the potent AhR antagonist 3'-methoxy-4'-nitroflavone (3'M4'NF) for in vivo activity. Wild type C57Bl/6 male mice were treated with solvent, benzo[a]pyrene (B[a]P; 150 mg/kg), or concurrently with B[a]P and 3'M4'NF (60 mg/kg; delivered as a split dose). Since B[a]P is bioactivated to genotoxic metabolites by AhR-regulated enzymes, we measured B[a]P-induced chromosomal damage in peripheral blood (i.e. micronuclei) to characterize the antagonistic potential of 3'M4'NF in vivo. The influence of AhR signal transduction was investigated further by challenging wild type and Ahr null allele mice with B[a]P with and without a 3'M4'NF co-treatment. The micronucleus data obtained from these experiments indicated that 3'M4'NF can attenuate the genotoxicity of B[a]P significantly. Since 3'M4'NF also protected Ahr null allele mice from B[a]P-induced genetic damage, it was apparent that AhR independent mechanisms contribute to the effects observed. However, as opposed to the protective effects observed with the micronucleus endpoint, histological observations and lethality data indicated that some B[a]P effects are enhanced by 3'M4'NF. Potentiated B[a]P toxicity may be explained by inhibition of basal and induced CYP1A1/2 activities. Both in vitro and in vivo data presented herein support this hypothesis. PMID- 10825464 TI - Inhibition of rabbit platelet aggregation by nucleoside 5'-alkylphosphates: correlation with inhibition of agonist-induced calcium influx. AB - We investigated the effects of uridine 5'-alkylphosphates on agonist-induced aggregation, increased intracellular calcium concentration [Ca(2+)](i), and Ca(2+) (Mn(2+)) influx in washed rabbit platelets. Uridine 5'-hexadecylphosphate (UMPC16) and uridine 5'-eicosylphosphate (UMPC20) at a concentration of 1 x 10( 5) M inhibited platelet aggregation induced by platelet-activating factor (PAF), thrombin, arachidonic acid, and ADP. UMPC16 did not cause significant interference in the binding of [(3)H-acetyl]PAF to platelets. The inhibition of PAF-induced platelet aggregation by UMPC16 was dependent upon the addition time; UMPC16 was ineffective at 60 sec when the extracellular calcium uptake reached the maximum level in PAF-stimulated platelets. Furthermore, UMPC16 inhibited guanosine 5'-O-(3-thiotriphosphate)-induced platelet aggregation but did not affect ionophore A23187- and calcium-independent agonist phorbol 12-myristate 13 acetate-induced platelet aggregation. UMPC16 markedly inhibited the Ca(2+) (Mn(2+)) influx induced by PAF and ADP, and partly inhibited the [Ca(2+)](i) increase induced by the receptor-mediated stimulation. On the other hand, UMPC16 did not affect the [Ca(2+)](i) increase and Ca(2+) (Mn(2+)) influx induced by ionomycin. These experiments suggest that inhibition of calcium influx associated with receptor-mediated platelet activation may be involved in the action of UMPC16. PMID- 10825465 TI - Role of NAD(P)H:quinone oxidoreductase 1 (DT diaphorase) in protection against quinone toxicity. AB - NQO1-/- mice, along with Chinese hamster ovary (CHO) cells, were used to determine the in vivo role of NAD(P)H:quinone oxidoreductase 1 (NQO1) in cellular protection against quinone cytotoxicity, membrane damage, DNA damage, and carcinogenicity. CHO cells permanently expressing various levels of cDNA-derived P450 reductase and NQO1 were produced. Treatment of CHO cells overexpressing P450 reductase with menadione, benzo[a]pyrene-3,6-quinone (BPQ), and benzoquinone led to increased cytotoxicity as compared with CHO cells expressing endogenous P450 reductase. In a similar experiment, overexpression of NQO1 significantly protected CHO cells against the cytotoxicity of these quinones. Knockout (NQO1-/ ) mice deficient in NQO1 protein and activity had been generated previously in our laboratory and were used in the present studies. Wild-type (NQO1+/+) and knockout (NQO1-/-) mice were given i.p. injections of menadione and BPQ, followed by analysis of membrane damage and DNA damage. Both menadione and BPQ induced lipid peroxidation in hepatic and non-hepatic tissues, indicating increased membrane damage. Exposure to BPQ also resulted in increased hepatic DNA adducts in NQO1-/- mice as compared with NQO1+/+ mice. The skin application of BPQ alone and BPQ + 12-O-tetradecanoylphorbol-13-acetate (TPA) failed to induce papillomas, or other lesions, for up to 50 weeks in either NQO1+/+ or NQO1-/- mice. The various results from CHO cells and NQO1-/- mice indicated that NQO1 protects against quinone-induced cytotoxicity, as well as DNA and membrane damage. The absence of BPQ-induced skin carcinogenicity in NQO1-/- mice may be related to the strain (C57BL/6) of mice used in the present study and/or due to poor BPQ absorption into the skin and/or due to detoxification of BPQ by cytosolic NRH:quinone oxidoreductase 2 (NQO2). PMID- 10825466 TI - Role of organic cation transporters in the renal secretion of nucleosides. AB - The mammalian kidney eliminates toxic substances from the body, in part via secretion by the organic cation transporters (OCT) or organic anion transporters. Nucleosides are nitrogenous heterocycles that are often secreted by human and other animal kidneys. Previous experiments have shown that 2'-deoxytubercidin (7 deazadeoxyadenosine, dTub) is secreted by the mouse kidney via a cimetidine sensitive OCT (Nelson et al., Biochem Pharmacol 32: 2323-2327, 1983). Experiments reported herein demonstrated that the cloned rat kidney rOCT1 transports dTub, cytosine arabinoside, 2-chlorodeoxyadenosine, and azidothymidine when expressed in the Xenopus laevis oocyte translation system. Although rOCT2 is 67% identical with rOCT1 in its amino acid sequence, rOCT2 does not mediate the uptake of these nucleosides. Uptake of dTub mediated by rOCT1 was pH-dependent in a manner suggesting that the positive charged moiety of dTub may be the true substrate. Protons acted as competitive inhibitors for the rOCT1-mediated uptake of dTub or tetraethylammonium (TEA), with K(i) values corresponding to a pH of about 6.1. TEA and dTub mutually inhibited the uptake of one another by rOCT1, competitively, with K(i) values approximately the same as their respective K(m) values. These findings suggest that protons, dTub, and TEA act at a common site on rOCT1, and that rOCT1 participates in the renal secretion of dTub and other nucleosides. PMID- 10825467 TI - Studies on the mechanism of the inhibition of human leukaemia cell growth by dietary isothiocyanates and their cysteine adducts in vitro. AB - The dietary isothiocyanates and cancer chemopreventive agents phenethyl isothiocyanate and allyl isothiocyanate and their cysteine conjugates inhibited the growth and induced apoptosis of human leukaemia HL60 (p53-) and human myeloblastic leukaemia-1 cells (p53+) in vitro. The median growth inhibitory concentration (GC(50)) values were in the range 1.49-3.22 microM in cultures with 10% serum. Isothiocyanates and cysteine conjugates had increased potency against HL60 cells in serum-free medium, with GC(50) values of 0.8-0. 9 microM. The potency of the compounds decreased with increased serum content of the medium, but that of the cysteine conjugates decreased more markedly. Growth inhibition and toxicity was characterised by either a rapid interaction of the isothiocyanate with the cells in the first hour of culture or exposure to isothiocyanate liberated from the cysteine conjugate in the initial 3 hr of culture, inhibition of macromolecule synthesis, and a commitment to apoptosis which developed in the initial 24 hr. Activities of caspase-3 and caspase-8 were increased during isothiocyanate-induced apoptosis, but caspase-1 activity was not. The general caspase inhibitor N-benzyloxycarbonyl-Val-Ala-Asp(OMe) fluoromethylketone and the specific caspase-8 inhibitor N-benzyloxycarbonyl-Ile Glu(OMe)-Thr-Asp(OMe)-fluoromethylketone inhibited apoptosis, but specific caspase-1 and caspase-3 inhibitors did not. The antiproliferative activities were limited by hydrolysis of the isothiocyanate. This suggests that caspase-8 has a critical role, and caspase-3 a supporting role, in isothiocyanate-induced apoptosis in which p53 is not an obligatory participant. Isothiocyanate-induced apoptosis may suppress the growth of preclinical tumours and contribute to the well-established decreased cancer incidence associated with a vegetable-rich diet. PMID- 10825468 TI - Association of tamoxifen biliary excretion rate with prior tamoxifen exposure and increased mdr1b expression. AB - ATPase transporter proteins are commonly found in the hepatocyte canalicular membrane. Some of these, in particular the multidrug resistance (mdr1b) gene, have been previously demonstrated to be inducible genes. In this study, we found that tamoxifen induced expression of the mdr1b gene in the liver up to 40-fold after 14 days' exposure to tamoxifen in the diet at a concentration of 420 ppm. As tamoxifen and its metabolites are primarily excreted into the bile, we investigated if the increased expression of mdr1b in the liver following tamoxifen exposure had any effect on its excretion in rats. We found that the excretion of tamoxifen and its metabolites into bile was increased from 8 +/- 1% to 51 +/- 18% (mean +/- SD) of an administered dose of 180 nmol/kg over a collection period of 3 hr in rats that had received tamoxifen (35 mg/kg) orally for 12 days (plus a 3-day rest) prior to the experiment. These data suggest that prolonged treatment with tamoxifen may result in lower serum and tumour concentrations, due to a self-mediated enhancement of excretion via mdr1b gene encoded P-glycoprotein. This may have implications for other drugs sharing the same route of excretion and co-administered with tamoxifen. PMID- 10825469 TI - Pharmacological modulation of the bystander effect in the herpes simplex virus thymidine kinase/ganciclovir gene therapy system: effects of dibutyryl adenosine 3',5'-cyclic monophosphate, alpha-glycyrrhetinic acid, and cytosine arabinoside. AB - The herpes simplex virus type 1 thymidine kinase (HSV1-tk) suicide gene/ganciclovir system was first applied to the treatment of glioblastoma tumors, but was hampered by the low gene transfection yield. Fortunately, the gap junction-dependent diffusion of phosphorylated ganciclovir metabolites from transfected cells to their neighbors proved to enhance the overall benefit of this strategy. However, as tumor cells are often gap junction-deficient, we sought to restore this property pharmacologically and hence to improve the efficacy of the treatment. We demonstrated that this approach was feasible in glioblastoma cells using dibutyryl adenosine 3',5'-cyclic monophosphate (cAMP) (100 microM) as a pharmacological inducer of gap junctions. alpha-Glycyrrhetinic acid (25 microM), on the other hand, strongly inhibited both gap junction mediated intercellular communication and the bystander effect, thus confirming the role of gap junctions in HSV-tk-mediated bystander killing. Using cytosine arabinoside as a growth inhibitor, we underlined the role of tumor cell proliferation in the sensitivity of HSV-tk-transfected cells to ganciclovir and demonstrated its correlation with the importance of the bystander effect. PMID- 10825470 TI - Actions of epoxyeicosatrienoic acid on large-conductance Ca(2+)-activated K(+) channels in pituitary GH(3) cells. AB - Epoxyeicosatrienoic acids (EETs), a family of cytochrome P450 epoxygenase metabolites of arachidonic acid, are believed to have an autocrine or paracrine role in the regulation of neurons or neuroendocrine cells. The effects of 14,15 EET on ionic currents were investigated in rat pituitary GH(3) cells. In the whole-cell configuration, 14,15-EET (3 microM) reversibly increased the amplitude of the Ca(2+)-activated K(+) current (I(K(Ca))). The 14, 15-EET-induced increase in I(K(Ca)) was unaffected in the presence of 10 microM thyrotropin-releasing hormone externally or 10 microM inositol trisphosphate in the recording pipette. In cells preincubated with pertussis toxin or herbimycin A, the 14, 15-EET induced increase in I(K(Ca)) was also not changed. In the inside-out configuration, 14,15-EET applied intracellularly did not change single-channel conductance, but did increase the opening probability of large-conductance Ca(2+) activated K(+) (BK(Ca)) channels. 14,15-EET (3 microM) shifted the activation curve of BK(Ca) channels to less positive membrane potential by approximately 15 mV. The change in the kinetic behavior of BK(Ca) channels caused by 14,15-EET is explained by a lengthening of open and a shortening of closed times. 14,15-EET increased the activity of BK(Ca) channels in a concentration-dependent manner with an EC(50) value of 1 microM. However, 14,15-EET did not affect the Ca(2+) sensitivity of BK(Ca) channels. The present study indicates that 14,15-EET is an opener of BK(Ca) channels in GH(3) cells and that the stimulatory effect of 14, 15-EET on these channels may, at least in part, contribute to the underlying cellular mechanisms by which EETs affect neuronal or neuroendocrine function. PMID- 10825471 TI - Improvement by aminoguanidine of insulin secretion from pancreatic islets grafted to syngeneic diabetic rats. AB - Prolonged hyperglycemia inhibits B-cell function by mechanisms that are largely unclarified. We investigated the involvement of advanced glycation end products (AGEs), using aminoguanidine as well as the AGE-breaking compound ALT-711 in a transplantation model. Islets from Wistar-Furth rats were transplanted under the kidney capsule of syngeneic streptozocin-diabetic recipients. Aminoguanidine was administered as 1 g/L in the drinking water. Graft-bearing kidneys were isolated and perfused to investigate insulin secretion, and grafts were excised to measure preproinsulin mRNA contents. In all transplants to diabetic rats, insulin responses to 27.8 mM glucose were abolished and aminoguanidine failed to correct this abnormality. However, aminoguanidine treatment for 8 weeks following transplantation increased preproinsulin mRNA contents of the grafts (P < 0.05). In addition, treatment with aminoguanidine enhanced the insulin secretory response to arginine (P < 0.05). Arginine-induced insulin secretion was also enhanced when aminoguanidine treatment was started after an initial 2-week implantation period rather than immediately after transplantation. On the other hand, treatment with ALT-711 (0.1 mg/kg by gavage) for 8 weeks completely failed to affect B-cell function of grafts, and ALT-711 was also ineffective under in vitro conditions. Our findings indicate that aminoguanidine effects in vivo are to a major extent not coupled to AGEs or nitric oxide synthetase inhibition, but possibly to oxidative modifications accomplished by the guanidine compound. PMID- 10825472 TI - Effectiveness of batimastat, a synthetic inhibitor of matrix metalloproteinases, in neutralizing local tissue damage induced by BaP1, a hemorrhagic metalloproteinase from the venom of the snake bothrops asper. AB - Batimastat (BB-94), a synthetic hydroxamate peptidomimetic matrix metalloproteinase inhibitor, was tested for its ability to inhibit proteolytic and toxic effects induced by BaP1, a 24-kDa hemorrhagic metalloproteinase isolated from the venom of Bothrops asper, the medically most important snake species in Central America and southern Mexico. Batimastat inhibited proteolytic activity on biotinylated casein, with anIC(50) of 80 nM. In addition, batimastat was effective in inhibiting hemorrhagic, dermonecrotic, and edema-forming activities of this metalloproteinase if incubated with the enzyme prior to the assays. When the inhibitor was administered i.m. at the site of the toxin injection without preincubation, rapidly after metalloproteinase administration, it totally abrogated the hemorrhagic and dermonecrotic effects of BaP1. Inhibition was less effective as the time lapse between toxin and batimastat injection increased, due to the extremely rapid development of BaP1-induced local tissue damage in this experimental model. On the other hand, batimastat was ineffective if administered by the i.p. route immediately after toxin injection. It is concluded that batimastat, and probably other synthetic metalloproteinase inhibitors, may become useful therapeutic tools aimed at the in situ inhibition of venom metalloproteinases, when injected at the site of the bite rapidly after envenomation. PMID- 10825473 TI - Tissue-specific gene expression of heme oxygenase-1 (HO-1) and non-specific delta aminolevulinate synthase (ALAS-N) in a rat model of septic multiple organ dysfunction syndrome. AB - Reactive oxygen species are thought to be involved in the pathogenesis of septic multiple organ dysfunction syndrome (MODS). It has been reported that heme oxygenase-1 (HO-1) (EC 1.14.99.3) is induced in septic animal models and is thought to confer protection against oxidative tissue injury. In this study, we examined changes in gene expression of HO-1 and non-specific delta aminolevulinate synthase (ALAS-N) (EC 2.3.1.37), the rate-limiting enzymes in heme catabolism and heme synthesis, respectively, after intraperitoneal administration of bacterial lipopolysaccharide (LPS) to rats. LPS treatment caused the elevation of body temperature, increases in white blood cell counts, and marked elevation of serum interleukin-6 levels associated with liver, lung, and kidney injuries, characteristic of septic MODS. LPS administration significantly induced HO-1 mRNA, protein, and enzyme activity in the liver, lung, and kidney. In contrast, ALAS-N mRNA was decreased rapidly in the liver, followed by an oscillating recovery pattern. Induction of hepatic HO-1 mRNA and rapid suppression of ALAS-N mRNA were likely the result of a rapid increase in hepatic free heme concentration as judged by the increase in heme saturation of tryptophan pyrrolase. In contrast to that in the liver, the ALAS-N mRNA level in the lung and kidney was increased significantly after LPS administration, suggesting a novel mechanism of ALAS-N regulation in these tissues. These findings suggest that HO-1 and ALAS-N mRNA are regulated in a tissue-specific manner in a rat model of septic MODS. PMID- 10825474 TI - Effect of phenobarbital on intralobular expression of CYP2B1/2 in livers of rats: difference in the expression between single and repetitive administrations. AB - Phenobarbital (PB) was shown to induce the major PB-inducible cytochrome P450 (CYP) isoforms, CYP2B1/2, in perivenular hepatocytes by a single injection, and in midzonal and periportal hepatocytes in addition to perivenular hepatocytes by injections of the same dosage once a day for 3 days in rat livers. The present study was undertaken to determine whether the spread of enzyme induction to midzonal and periportal hepatocytes is caused by the increase in total dose of the drug by repetitive injections or by the repetitive injections of the drug themselves. Male adult rats were administered PB by a single injection (80 mg/kg) or repetitive injections (20 mg/kg once a day for 4 days; a total dose of 80 mg/kg), and the molar content of CYP2B1/2 was measured by quantitative immunohistochemistry in the cytoplasm of perivenular, midzonal, and periportal hepatocytes. In addition, the molar content of total CYP in the cytoplasm was measured by microphotometry, and the expression of CYP2B2 mRNA was examined by in situ hybridization. When animals received the single injection, the isoforms and CYP2B2 mRNA increased markedly in perivenular hepatocytes, increased somewhat in midzonal hepatocytes, and remained unchanged in periportal hepatocytes. If animals received the repetitive injections, however, although the isoforms and the mRNA increased markedly in perivenular hepatocytes, they also increased markedly in midzonal hepatocytes and somewhat in periportal hepatocytes. These findings demonstrated that the enlargement of the sublobular area in which induction of the isoforms occurred was caused by the repetitive injections of PB themselves. PMID- 10825475 TI - Neurotrophin-receptor immunoreactive neurons in mesopontine regions involved in the control of behavioral states. AB - The microinjection of nerve growth factor (NGF) and neurotrophin-3 (NT-3) into the rostral pontine tegmentum of adult cats rapidly induces long-lasting episodes of rapid eye movement (REM) sleep [J. Yamuy, F.R. Morales, M.H. Chase, Induction of rapid eye movement sleep by the microinjection of nerve growth factor into the pontine reticular formation of the cat, Neuroscience 66 (1995) 9-13]. Because this effect may be mediated by neurotrophin receptors, we sought to determine the distribution of neurons that contain low- and high-affinity neurotrophin receptors in regions of the feline pons and mesencephalon which are involved in the generation of REM sleep as well as neuronal groups that are involved in the control of REM sleep-related patterns of physiological activity. Using antibodies directed against p75, trkA, trkB and trkC, immunolabeled neurons were present in the latero-dorsal and pedunculo-pontine tegmental nuclei, the peribrachial nuclei, medial and lateral pontine reticular formation, the raphe nuclei, and the locus coeruleus. Giant reticular cells and large neurons in the mesencephalic trigeminal nucleus were immunoreactive for p75 and all trk receptors. Neurons that were devoid of neurotrophin-receptor immunoreactivity were intermingled with immunostained neurons in all explored structures. Thus, both low- and high affinity neurotrophin receptors are conspicuously present in neurons located in mesopontine regions of adult cats. These data underscore the importance of neurotrophin-induced trophic actions on mesopontine neurons. Furthermore, the results support the hypothesis that NGF and NT-3 may modulate the electrical activity of neurons in the rostral pontine tegmentum that are responsible for the generation of REM sleep by acting on one or more of the neurotrophin receptors. PMID- 10825476 TI - Effects of neutralizing antibodies to TNF-alpha on pain-related behavior and nerve regeneration in mice with chronic constriction injury. AB - Inhibition of proinflammatory cytokines reduces hyperalgesia in animal models of painful neuropathy. We set out to investigate the consequences of this treatment for nerve regeneration. Here we examined the sequels of epineurial application of neutralizing antibodies to tumor necrosis factor-alpha (TNF) in chronic constriction injury (CCI) of the sciatic nerve in C57/BL 6 mice. The mice were tested behaviorally for manifestations of thermal hyperalgesia and mechanical allodynia. Nerve regeneration was assessed by morphometry of myelinated nerve fibers in the sciatic nerve and of the epidermal innervation density in the glabrous skin of the hindpaws. Antibodies to TNF reduced thermal hyperalgesia and mechanical allodynia after CCI. Myelinated fiber density in the sciatic nerve was reduced to 30% of normal on day 7 after surgery, and reached 60% on day 45, with no difference between antibody-treated and untreated animals. Epidermal innervation density as shown by PGP 9.5 and CGRP immunohistochemistry was reduced to 25-47% at both time points after CCI, again without differences between antibody treated and untreated mice. Myelinated fiber density but not epidermal innervation density was correlated to thermal and mechanical withdrawal thresholds. We conclude that neutralization of endoneurial TNF attenuates pain related behavior but has no effect on nerve regeneration. Furthermore, the number of epidermal nerve fibers is not relevant to the magnitude of behavioral hyperalgesia in CCI. PMID- 10825477 TI - Preconditioning-induced neuroprotection is mediated by reactive oxygen species. AB - The current study was performed to determine the role of reactive oxygen species (ROS) in preconditioning against different forms of neuronal damage. Primary cultures of chick embryonic neurons were treated with either FeSO(4) (100 microM; 15 min) to generate hydroxyl radicals or xanthine/xanthinoxidase (10 microM/0.5 mU ml(-1); 15 min; =X/XO (pre)) to produce superoxide radicals. Both stimuli moderately enhanced ROS formation as measured by fluorescence microscopy. This preconditioning significantly protected the neurons against subsequent glutamate (1 mM)-induced excitotoxic damage, staurosporine (200 nM)-induced neuronal apoptosis and oxidative damage caused by exposure to xanthine/xanthinoxidase (500 microM/5 mU ml(-1); 1 h; =X/XO (dam)). The antioxidants vitamin E (10 microM) and 2-OH-estradiol (1 microM), present during the 15-min preconditioning period, completely abolished the protective effect of X/XO (pre). Furthermore, glutamate, staurosporine or X/XO (dam) markedly enhanced oxygen radical formation. Preceding preconditioning by mild ROS stimulation with X/XO (pre) or Fe(2+) reduced this oxygen radical burst. Again, the effect of X/XO (pre) could be blocked by coadministration of vitamin E or 2-OH-estradiol. However, the FeSO(4)-mediated preconditioning was not abolished by the radical scavengers. To address this phenomenon, the effect of vitamin E and 2-OH-estradiol on Fe(2+)- and X/XO (pre) induced ROS formation kinetics within the 15 min of preconditioning was monitored. The moderate rise of intracellular ROS content during preconditioning was only reduced permanently by the antioxidants, when the neurons were treated with X/XO (pre), but not when Fe(2+) was used. Thus, an immediate and constant radical scavenging seems to be indispensable to abolish the ROS-induced neuronal preconditioning. The current results indicate that preconditioning by moderate ROS-stimulation protects cultured neurons against different damaging agents and prevents against the subsequent massive oxygen radical formation. PMID- 10825478 TI - Overexpression of human alpha-synuclein causes dopamine neuron death in rat primary culture and immortalized mesencephalon-derived cells. AB - Parkinson's disease (PD) is a neurodegenerative disorder characterized by the appearance of intracytoplasmic inclusions called Lewy bodies (LB) in dopamine neurons in the substantia nigra and the progressive loss of these neurons. Recently, mutations in the alpha-synuclein gene have been identified in early onset familial PD, and alpha-synuclein has been shown to be a major component of LB in all patients. Yet, the pathophysiological function of alpha-synuclein remains unknown. In this report, we have investigated the toxic effects of adenovirus-mediated alpha-synuclein overexpression on dopamine neurons in rat primary mesencephalic cultures and in a rat dopaminergic cell line - the large T antigen immortalized, mesencephalon-derived 1RB3AN27 (N27). Adenovirus-transduced cultures showed high-level expression of alpha-synuclein within the cells. Overexpression of human mutant alpha-synuclein (Ala(53)Thr) selectively induced apoptotic programmed cell death of primary dopamine neurons as well as N27 cells. The mutant protein also potentiated the neurotoxicity of 6-hydroxydopamine (6 OHDA). By contrast, overexpression of wild-type human alpha-synuclein was not directly neurotoxic but did increase cell death after 6-OHDA. Overexpression of wild-type rat alpha-synuclein had no effect on dopamine cell survival or 6-OHDA neurotoxicity. These results indicate that overexpression of human mutant alpha synuclein directly leads to dopamine neuron death, and overexpression of either human mutant or human wild-type alpha-synuclein renders dopamine neurons more vulnerable to neurotoxic insults. PMID- 10825479 TI - Nucleus accumbens cell firing during maintenance, extinction, and reinstatement of cocaine self-administration behavior in rats. AB - Electrophysiological recording procedures were used to examine nucleus accumbens (Acb) cell firing in rats (N=13) during cocaine self-administration sessions consisting of three phases. In phase one (maintenance), each lever press resulted in an intravenous cocaine infusion (0.33 mg, 6 s) paired with a tone-houselight stimulus (20 s). Of 144 Acb cells recorded during maintenance, 39 neurons (27%) exhibited phasic firing relative to the cocaine-reinforced response [4-8]. Briefly, Acb neurons showed increases in firing rate within seconds preceding the reinforced response and/or changes (increases or decreases) in activity within seconds following response completion. In phase two (extinction), saline was substituted for cocaine in the task. Results indicated that cells displaying exclusively anticipatory discharges during maintenance exhibited similar phasic activity during extinction. However, neurons that displayed post-response activity during the maintenance phase typically showed significant attenuation of phasic firing rates during extinction. After 30 min of no responding, animals were 'primed' with an intravenous infusion of cocaine, and self-administration was reestablished during phase three (reinstatement). Results showed that pre response discharge patterns remained relatively intact while post-response cells typically exhibited a partial recovery of phasic activity. Similar findings were observed during other extinction experiments in which the stimulus only was removed (CS extinction). These findings support the notion that specific factors operating within the self-administration context differentially control pre- versus post-response discharge patterns of Acb neurons. PMID- 10825480 TI - Calretinin-containing axons and neurons are resistant to an intrastriatal 6 hydroxydopamine lesion. AB - Relative preservation of dopaminergic axons in patches and a subcallosal layer was observed in the dorsal, lateral and caudal striatum 4 weeks after intrastriatal injection of 6-hydroxydopamine (6-OHDA), a neurotoxin selective for catecholaminergic neurons. Since calcium binding proteins are reported to provide neuroprotective influence in neurons, differences in the distribution of the calcium binding proteins might be related to the different vulnerabilities of dopaminergic neurons and axons to neurotoxins. To address this possibility, we characterized patches of relatively dense tyrosine hydroxylase-immunoreactive (TH IR) axons in intrastriatal 6-OHDA lesioned rats, focusing on two calcium binding proteins, calbindin (CB) and calretinin (CR). The patches and subcallosal layer of preserved dopaminergic axons in the striatum of rats lesioned with 6-OHDA contained CR, a 31-kDa calcium-binding protein, but interestingly not CB. Dopaminergic neurons containing CR in the substantia nigra pars compacta (SNpc) were relatively spared compared to those that did not contain CR. Taken together, our data indicate that dopaminergic axons and neurons containing CR in the nigrostriatal pathway are more resistant to 6-OHDA lesion than those that do not contain CR. PMID- 10825481 TI - Muscarinic receptors modulate intracellular calcium level in chick sensory neurons. AB - In the present work we have studied the variation of intracellular calcium levels induced by muscarinic agonists in chick dorsal root ganglia neurons. Muscarinic agonists such as muscarine and oxotremorine cause an increase of intracellular calcium levels in fura-2AM-loaded DRG neurons of E18 chick embryos. This increase was abolished following treatment with 1 microM atropine but not by 1 microM mecamylamine, indicating that the observed intracellular calcium increase, was dependent on muscarinic receptor activation. Stimulation in absence of external calcium or pre-incubation of the DRG cultures with thapsigargin or Mn(2+) demonstrated that [Ca(2+)](i) increase is mainly due to its release from intracellular stores. The use of selective antagonists of muscarinic receptor subtypes also indicated that M(1) and to a lesser extent M(3) receptor subtypes are responsible for the observed intracellular calcium mobilization. Finally pre treatment of DRG cultures with pertussis toxin showed that the variation of [Ca(2+)](i) levels was dependent on PTX-insensitive G-protein. Moreover muscarinic agonists induce in DRG also the increase of IPs level, suggesting that the variations of intracellular calcium levels may be due at least in part to the activation of the phosphoinositide transduction pathway. In conclusion the reported observations demonstrate the activity of muscarinic receptors in sensory neurons, suggesting a functional role for acetylcholine in sensory transduction. PMID- 10825482 TI - Regulation of high-affinity glutamate uptake activity in Bergmann glia cells by glutamate. AB - The effect of glutamate receptor activation on the high-affinity sodium-dependent glutamate transport expressed in chick Bergmann glia cells was examined. Pre exposure to glutamate produced a time- and dose-dependent decrease in 3H-labeled D-aspartate uptake. This effect could not be reproduced by selective glutamate receptor agonists. Furthermore, it was insensitive to both ionotropic and metabotropic glutamate receptor antagonists. Replacement of extracellular sodium ions with choline in the preincubation media, abolished the reduction of the uptake. When the cells were pre-exposed to competitive transportable inhibitors of the transporter, such as D-aspartate, DL-threo-hydroxyaspartate (DL-THA), and aspartate-beta-hydroxamate (ABH), the glutamate effect was mimicked. From saturation experiments, it was found that the reduction on the uptake, after glutamate treatment, is related to an increase in K(m). Interestingly, the effect is blocked by staurosporine, a Ca(2+)/diacylglycerol-dependent protein kinase (PKC) inhibitor. The present findings suggest that glutamate regulates its transport in a non-receptor fashion, a phenomena that is most probably linked to changes induced by the translocation process of the substrate through the transporter. PMID- 10825483 TI - CRF and urocortin decreased brain stimulation reward in the rat: reversal by a CRF receptor antagonist. AB - The present studies were designed to investigate the effects of corticotropin releasing factor (CRF) receptor activation and antagonism on intracranial self stimulation (ICSS) reward using a discrete-trial current-intensity threshold procedure. Bipolar electrodes were implanted in the lateral hypothalamus, and cannula guides were implanted above the lateral ventricle of male Wistar rats. Dose-effect functions were established for the effects on ICSS of the competitive CRF receptor agonist h/rCRF (0-5.0 microg, i.c.v. ), the CRF receptor agonist urocortin (0-5.0 microg, i.c.v.), and the CRF receptor antagonist [D-Phe(12), Nle(21,38), C(alpha) MeLeu(37)] h/rCRF(12-41) (0-5.0 microg, i.c.v.). Administration of h/rCRF or urocortin dose-dependently elevated ICSS thresholds without altering performance measures (latencies to respond to stimulation, extra and time-out responses). CRF was more potent than urocortin in terms of threshold dose-effects on ICSS thresholds compared to vehicle. Despite these apparent potency differences, percent effect sizes on ICSS thresholds were comparable at the highest doses of both peptides. In contrast to the significant threshold elevation effects of CRF and urocortin, the competitive CRF antagonist D-Phe CRF(12-41) had no effect on ICSS thresholds or performance measures. To determine the neuropharmacological specificity of the effect of CRF on brain stimulation reward, D-Phe CRF(12-41) was used to antagonize CRF-induced threshold elevations. Pretreatment with either the 5.0- or 10.0-microg doses of D-Phe CRF(12-41) effectively blocked CRF-induced reward threshold elevations (3.0 microg) without affecting other ICSS performance measures. These results indicate that CRF neurotransmission can modulate ICSS reward processes. PMID- 10825484 TI - Deafferentation-induced changes in the olfactory bulb of adult zebrafish. AB - The influence of the olfactory organ on maintenance of olfactory bulb structure was examined in zebrafish, using peripheral deafferentation. This fish provides a model in which the olfactory organ is easily accessible for removal, the animals easily survive the surgery, and the olfactory bulbs are small enough to allow rigorous analysis of the resulting effects. Unilateral olfactory organ ablations were performed on anesthetized adult zebrafish using a small-vessel cautery iron. Fish were allowed to survive for 1, 3, or 6 weeks following the procedure. Analysis of deafferented animals revealed that most, if not all, of the olfactory organ was missing on the ablated side, and the structure did not regenerate. The morphology of the olfactory bulb was affected notably by the removal of its primary afferent innervation. The olfactory nerve layer was diminished at 1 week and absent by 3 weeks post-deafferentation. At all of the survival times the deafferented bulb appeared significantly smaller at the gross level, and there was a statistically significant effect on bulb size and cell number after 6 weeks. Tyrosine hydroxylase expression, as revealed by immunohistochemistry, was decreased noticeably on the ablated side. In conclusion, the olfactory organ is important in the preservation of normal olfactory bulb anatomy and neurochemistry in adult zebrafish. Thus, the influence of the periphery does not end with the formation of the mature olfactory bulb. PMID- 10825485 TI - Nitric oxide donor-induced increase in permeability of the blood-brain barrier. AB - The goal of the present study was to determine the effect of nitric oxide (NO) donors on the permeability of the blood-brain barrier in vivo. We examined the pial microcirculation in rats using intravital fluorescence microscopy. Permeability of the blood-brain barrier was quantitated by calculating the clearance of fluorescent-labeled dextran (M(w)=10000 Da; FITC-dextran-10K) during suffusion with vehicle, S-nitroso-N-acetylpenicillamine (SNAP; 100 microM) and 3 morpholinosydnonimin (SIN-1; 100 microM). In addition, we examined changes in arteriolar diameter during suffusion with vehicle, SNAP and SIN-1. During suffusion with vehicle, clearance of FITC-dextran-10K from pial vessels and diameter of pial arterioles remained relatively constant during the experimental period. In contrast, suffusion with SNAP or SIN-1 markedly increased clearance of FITC-dextran-10K from the cerebral microcirculation and produced a rapid, sustained dilatation of pial arterioles. Thus, NO donors increase the permeability of the blood-brain barrier and produce pronounced dilatation of cerebral arterioles. In light of evidence suggesting that NO donors may produce their effect by the simultaneous release of NO and superoxide anion to form peroxynitrite, we elected to examine the role of superoxide anion in increases in permeability of the blood-brain barrier in response to SNAP and SIN-1. We found that suffusion with tiron (1 mM) did not alter basal permeability of the blood brain barrier, but significantly inhibited increases in permeability of the blood brain barrier in response to SNAP and SIN-1. In addition, tiron did not alter baseline diameter of cerebral arterioles, or SNAP- and SIN-1-induced cerebrovasodilatation. The findings of the present study suggest that NO donors produce an increase in permeability of the blood-brain barrier which appears to be related to the presence of NO and superoxide anion, to presumably form peroxynitrite. We suggest that increases in NO formation observed during brain trauma may contribute to disruption of the blood-brain barrier. PMID- 10825486 TI - Genetic variability in morphine sensitivity and tolerance between different strains of rats. AB - The development of tolerance, the sensitivity to morphine and the effective morphine plasma concentrations have been studied in Sprague-Dawley (SD-U) and Wistar (W) rats. Daily administration of morphine (10 mg/kg/12 h for 9 days) in W rats produced a reduction in morphine antinociception from day 1 (12+/-0 s) to day 9 (6.7+/-1. 9 s). Morphine antinociception in the SD-U rats did not change over the period of treatment. Naloxone abolished the antinociception of morphine in both opiate naive and chronically treated SD-U rats. The pharmacokinetic parameters of morphine and morphine-3-glucuronide did not differ significantly between strains. Both naive and chronically treated SD-U rats required smaller doses of morphine than W rats to obtain a maximum antinociceptive effect. Plasma concentrations following administration of the same dose of morphine, did not differ between strains or days of treatment. The range of morphine concentrations required to obtain a maximum effect were lower in SD-U rats, both on day 1 and day 8 when compared to W rats. These results show differences between the two strains with regard to both morphine sensitivity and development of tolerance, whilst also suggesting that the differences do not have a kinetic basis. PMID- 10825487 TI - Sampling extracellular aspartate, glutamate and gamma-aminobutyric acid in striate cortex of awake cat by in vivo microdialysis: surgical and methodological aspects. AB - A method which permits repeated microdialysis in the cortical layers of area 17 of the awake cat is described. Under visual control through a surgical microscope and using a stereotactic instrument, four probe guides are permanently implanted in area 17 of one hemisphere of the anesthetized animal and two fixation bars are mounted on the skull to allow fixation of the cat in a stereotactic frame. The implantation of four probe guides in the same hemisphere allows simultaneous sampling from different cortical regions serving different parts of the visual field. A removable transparent cover protects the probe guides. After recovery from surgery the awake cats are trained to adapt to a fixation of 5 h in a stereotaxic apparatus. Once adapted to that situation, the cats are ready for microdialysis experiments without anesthesia. The day of the experiment, the awake animal was fixed in the stereotactic frame and the probes inserted into the guides. To test the validity of the method, the basal efflux and the depolarization efflux, triggered by the addition of 65 mM K(+) to the artificial cerebrospinal fluid, of the amino acids aspartate, glutamate and gamma aminobutyric acid are measured by two HPLC-electrochemical detection methods. The exact localization of the probes and the reaction of the surrounding tissue is studied using immunocytochemistry for glutamate and glial fibrilary acidic protein. Our neurochemical and morphological results suggest the feasibility of multiple and repeated probe insertions for microdialysis experiments in the cerebral cortex of awake and behaving cat. This method provides a new tool to investigate the cortical plasticity. PMID- 10825488 TI - Cocaine- and amphetamine-regulated transcript, glucagon-like peptide-1 and corticotrophin releasing factor inhibit feeding via agouti-related protein independent pathways in the rat. AB - The melanocortin-4 receptor (MC4-R) appears to be an important downstream mediator of the action of leptin. We examined to what extent the anorectic effects of cocaine- and amphetamine-regulated transcript (CART), glucagon-like peptide-1 (GLP-1) and corticotrophin releasing factor (CRF) might be mediated via MC4-R. alpha-Melanocyte stimulating hormone (alpha-MSH), the MC4-R agonist, administered intracerebroventricularly (ICV) at a dose of 1 nmol reduced food intake by approximately half. Agouti-related protein (Agrp) (83-132), a biologically active fragment of the endogenous MC4-R antagonist, administered ICV at a dose of 1 nmol completely blocked the anorectic effect of 1 nmol alpha-MSH. CART (55-102) (0.2 nmol), GLP-1 (3 nmol) and CRF (0.3 nmol) produced a reduction in feeding of approximately the same magnitude as 1 nmol alpha-MSH. Agrp (83-132) (1 nmol) administered ICV did not block the anorectic effects of CART (55-102) (1 h food intake, 0.2 nmol CART (55-102), 2.7+/-0.8 g vs. CART (55-102)+Agrp (83 132), 2.6+/-0.6 g, P=0.87; saline control 5.4+/-0.3 g, P<0.001 vs. both groups). Agrp (83-132) also did not block the anorectic effects of GLP-1 or CRF (1 h food intake, 0.3 nmol CRF, 0.7+/-0.3 g vs. CRF+Agrp (83-132), 0.7+/-0.3 g, P=0.91; 3 nmol GLP-1, 1.9+/-0.4 g vs. GLP-1+Agrp (83-132), 1.1+/-0. 5 g, P=0.23; saline control 5.0+/-0.6 g, P<0.001 vs. all four groups). Thus, as previous data suggests, GLP-1 and CRF do not appear to reduce food intake predominantly via MC4 R, we here demonstrate for the first time that CART, in addition to GLP-1 and CRF primarily acts via Agrp independent pathways. PMID- 10825489 TI - c-Fos-like immunoreactivity in the brainstem following gastric loads of various chemical solutions in rats. AB - The distribution of c-Fos-like immunoreactivity (c-FLI) in the lower brainstem especially in the area postrema (AP), nucleus of the tractus solitarius (NTS) and parabrachial nucleus (PBN) was examined following gastric loads of various chemical solutions in rats. An aliquot of 7.5 ml of each stimulus was intragastrically infused, and c-FLI was detected. The most remarkable c-FLI was induced by LiCl, lactose and ethanol which are known to be effective unconditioned stimuli in conditioned taste aversions. Polycose and disaccharides such as sucrose and maltose induced more c-FLI than monosaccharides such as glucose, fructose and galactose. Relatively low levels of c-FLI were observed for other sweeteners such as saccharin, glycine and alanine, and other basic taste stimuli such as NaCl, HCl, quinine and umami substances. Each stimulus induced a similar proportion of c-FLI among the subnuclei of the NTS, but not in the PBN, where chemicals effective in inducing conditioned taste aversions elicited stronger c-FLI in the external lateral subnucleus, and those in inducing conditioned taste preferences such as Polycose and glucose elicited stronger c FLI in the dorsal lateral subnucleus. Vagotomy reduced c-FLI to about 50% for LiCl stimulation and to about 30% for sucrose stimulation, suggesting that LiCl has a larger proportion of extravagal inputs than sucrose. PMID- 10825490 TI - Comparison of c-fos-like immunoreactivity in the brainstem following intraoral and intragastric infusions of chemical solutions in rats. AB - To examine whether the activation of brainstem neurons during ingestion is due to orosensory afferents or post-ingestive factors, neuronal activation in response to intraoral and intragastric infusions of taste stimuli was compared in the area postrema (AP), nucleus tractus solitarius (NTS) and parabrachial nucleus (PBN) by the c-fos immunohistochemical method. An aliquot (7.5 ml) of 0.5 M sucrose, 5 mM sodium saccharin, 1 mM quinine hydrochloride and distilled water was delivered into the oral cavity or the stomach in each rat, which had been deprived of water and food overnight. Water induced little c-Fos-like immunoreactivity (c-FLI), but both intraoral and intragastric infusions of sucrose, but not non-caloric saccharin, induced strong c-FLI in the AP, caudal NTS and the external lateral subnucleus of the rostral PBN, suggesting that these areas receive general visceral inputs. Other areas in the NTS and PBN may receive gustatory inputs since more dominant c-FLI was detected by intraoral rather than intragastric infusions of the stimuli. Functional segregation of neurons reflecting qualitative and hedonic aspects of sweeteners (sucrose and saccharin) and bitter tasting substance (quinine) was suggested in the PBN, but less evident in the NTS. These results indicate that c-fos induction in brainstem neurons during ingestion reflects gustatory inputs and postingestional factors depending on the kind of food ingested. PMID- 10825491 TI - Ultradian oscillations in cranial thermoregulation and electroencephalographic slow-wave activity during sleep are abnormal in humans with annual winter depression. AB - The level of core body, and presumably brain temperature during sleep varies with clinical state in patients with seasonal affective disorder (SAD), becoming elevated during winter depression and lowered during clinical remission induced by either light treatment or summer. During sleep, brain temperatures are in part determined by the level of brain cooling activity, which may be reflected by facial skin temperatures. In many animals, the level of brain cooling activity oscillates across the NREM-REM sleep cycle. Facial skin temperatures during sleep in patients with winter depression are abnormally low and uncorrelated with rectal temperatures, although their relationship to EEG-defined sleep stages remains unknown. We therefore measured the sleep EEG, core body and facial skin temperatures in 23 patients with winter depression and 23 healthy controls, and tested the hypothesis that ultradian oscillations in facial skin temperatures exist in humans and are abnormal in patients with winter depression. We found that facial skin temperatures oscillated significantly across the NREM-REM sleep cycle, and were again significantly lower and uncorrelated with rectal temperatures in patients with winter depression. Mean slow-wave activity and NREM episode duration were significantly greater in patients with winter depression, whereas the intraepisodic dynamics of slow-wave activity were normal in patients with winter depression. These results suggest that brain cooling activity oscillates in an ultradian manner during sleep in humans and is reduced during winter depression, and provide additional support for the hypothesis that brain temperatures are elevated during winter depression. PMID- 10825492 TI - Responses of rat spinal neurones to natural and electrical stimulation of colonic afferents: effect of inflammation. AB - Single unit electrical activity has been recorded from 107 neurones excited by electrical stimulation of the pelvic nerve in or around lamina X of the L6-S1 spinal cord in anaesthetised rats. Responses to colorectal distension (CRD; 30 s, 5-80 mmHg) and to somatic electrical and mechanical stimulation were characterised. Of 107 neurones excited by pelvic nerve stimulation, 58 (54%) were affected by CRD: 46 neurones were excited (39 with a sustained response and 7 with an on-off response) and 12 neurones were inhibited. The vast majority of the neurones affected by CRD (54/58) had nociceptive somatic receptive fields. Neurones excited by CRD showed graded stimulus response functions in the noxious range (20-80 mmHg), except for two neurones which only encoded stimulus intensity below 20 mmHg. Neurones inhibited by CRD had significantly larger somatic receptive fields, and more superficial recording sites than those excited by CRD. A group of 12 neurones with sustained excitatory responses to CRD were characterised before and 45 min after intracolonic instillation of 1 ml 0.6% acetic acid. Colon inflammation provoked a significant increase in responses to CRD and to pelvic nerve stimulation (n=12), but no significant change in responses to pinch of their somatic receptive field (n=10). We conclude that of these neurones, the population with excitatory sustained responses to CRD are those likely responsible for processing information leading to acute pain sensations from the colon, and also show central sensitisation after colon inflammation, suggesting they play an important role in development of colonic hyperalgesia. PMID- 10825493 TI - Tactile directional sensibility: peripheral neural mechanisms in man. AB - Tactile directional sensibility, i.e. the ability to tell the direction of an object's motion across the skin, is an easily observed sensory function that is highly sensitive to disturbances of the somatosensory system. Based on previous psychophysical experiments on healthy subjects it was concluded that directional sensibility depends on two kinds of information from cutaneous mechanoreceptors; spatio-temporal information and information about friction-induced changes in skin stretch. In the present study responses to similar probe movements as in the psychophysical experiments were recorded from human single mechanoreceptors in the forearm skin. All slowly adapting type 2 (SA2) units were spontaneously active, and with increasing force of friction their discharge rates were modified by probe movements at increasing distances from the Ruffini end-organ, reflecting the high stretch-sensitivity of these units. Slowly adapting type 1 (SA1) and field units responded to the moving probe within well-defined skin areas directly overlying the individual receptor terminals, and compared to the SA2 units their response properties were less dependent on the force of friction. The results suggest that SA1 and field units have the capacity to signal spatio-temporal information, whereas a population of SA2 units have the capacity to signal direction-specific information about changes in lateral skin stretch. PMID- 10825494 TI - Afferent high strength tetanizations favour potentiation of the NMDA vs. AMPA receptor-mediated component of field EPSP in CA1 hippocampal slices of rats. AB - Long-term potentiation (LTP) of alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid (AMPA) and N-methyl-D-aspartate (NMDA) receptor-mediated components of 'dual-component' field excitatory postsynaptic potentials (fEPSP-A and fEPSP-N) was studied in the CA1 stratum radiatum in hippocampal slices of rats. Relative degrees of LTP of these fEPSP components were compared for tetanizations with low and high strengths. Magnitudes of fEPSP-A and fEPSP-N were estimated in parallel with a least-square fitting of a short-latent (0.1-8.8 ms) fragment of evoked responses by a weighted sum of 'basic' fEPSP-A and fEPSP-N, obtained during a short preliminary application of d-2-amino-5-phosphonovalerate (APV). We found that low-strength tetanizations selectively potentiated fEPSP-A, while high strength tetanizations potentiated both fEPSP components. These results demonstrate in the experiments with parallel measurements of fEPSP-A and fEPSP-N that LTP of these components differ depending on the strength of afferent tetanization. Unequal potentiation of the commissural-collateral and excitatory local-circuit synapses, which presumably contain different amounts of the AMPA and NMDA receptors, is discussed as the most probable explanation for these results. PMID- 10825495 TI - Effects of a unilateral stereotaxic injection of Tinuvin 123 into the substantia nigra on the nigrostriatal dopaminergic pathway in the rat. AB - Tinuvin 123, a compound used in the manufacture of plastics, has recently been suggested to possibly cause Parkinson's disease (PD). Herein, we revisited this issue by assessing the effect of Tinuvin 123 on dopaminergic neurons of the substantia nigra following its stereotaxic injection in the rat. Twenty-one days post unilateral stereotaxic injection of Tinuvin 123, systemic injection of both apomorphine and amphetamine caused rotations toward the side of the lesion in these rats. Tinuvin 123 produced a small to moderate dose-dependent reduction in striatal levels of dopamine and metabolites on the side of the lesion. This compound also produced dramatic cell loss in the substantia nigra on the side of the lesion. However, the loss of cells lacked the phenotypic specificity for tyrosine hydroxylase (TH)-positive neurons that is expected with a dopaminergic neurotoxin. Indeed, aside from a robust glial reaction, both TH-positive and glutamic acid dehydrogenase (GAD)-positive neurons were destroyed, and near the site of the injection, there was complete tissue destruction. This study indicates that, using this mode of injection, Tinuvin 123 exerts a dramatic tissue toxicity without any evidence of specificity for dopaminergic neurons. Thus, our data argues against a role for Tinuvin 123 as an environmental toxin causing a clinical condition characterized by the selective loss of dopaminergic neurons as seen in PD. PMID- 10825496 TI - Antioxidant-rich diets improve cerebellar physiology and motor learning in aged rats. AB - The free radical theory of aging predicts that reactive oxygen species are involved in the decline in function associated with aging. The present paper reports that diets supplemented with either spinach, strawberries or blueberries, nutritional sources of antioxidants, reverse age-induced declines in beta adrenergic receptor function in cerebellar Purkinje neurons measured using electrophysiological techniques. In addition the spinach diet improved learning on a runway motor task, previously shown to be modulated by cerebellar norepinephrine. Motor learning is important for adaptation to changes in the environment and is thus critical for rehabilitation following stroke, spinal cord injury, and the onset of some neurodegenerative diseases. These data are the first to indicate that age-related deficits in motor learning and memory can be reversed with nutritional interventions. PMID- 10825497 TI - Membrane depolarization-mediated survival of sympathetic neurons occurs through both phosphatidylinositol 3-kinase- and CaM kinase II-dependent pathways. AB - It has been well established that the NGF-mediated survival of sympathetic neurons in culture occurs through the phosphatidylinositol (PI) 3-kinase/Akt dependent pathway. In contrast, the mechanism by which membrane depolarization promotes neuronal survival independently of NGF remains unresolved. Here we show that LY294002, a specific inhibitor of PI 3-kinase, induced cell death of sympathetic neurons under depolarizing conditions with elevated K(+) (IC(50)= approximately 30 microM). Interestingly, lower concentrations of this agent (< or =10 microM) were sufficient to suppress Akt phosphorylation at Ser-473, a putative downstream target of PI 3-kinase, under these conditions. We also show that KN-62, a specific inhibitor of Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) suppressed depolarization-mediated survival in a does-dependent manner (IC(50)= approximately 2 microM) that paralleled attenuation of sustained levels of intracellular Ca(2+) evoked by depolarization. This IC(50) value is greater than that for CaMKII ( approximately 0.8 microM). These findings led us to hypothesize that depolarization-mediated survival occurs through both the PI 3 kinase/Akt and the CaMKII pathways. Indeed, combined treatment with LY294002 (25 microM) and KN-62 (0.5 microM) dramatically abolished depolarization-mediated survival, whereas each alone did not significantly attenuate it. Under these conditions, KN-62 neither impaired sustained levels of intracellular Ca(2+), nor inhibited the phosphorylation of Akt. It is thus likely that PI 3-kinase and CaMKII independently promote the membrane depolarization-mediated survival of sympathetic neurons in culture. PMID- 10825498 TI - Ultrastructural characteristics and conduction velocity of olfactory receptor neuron axons in the olfactory marker protein-null mouse. AB - Olfactory receptor neuron (ORN) axon diameters and the conduction velocity of the compound action potential along ORN axons were studied in olfactory marker protein (OMP)-null mice and genotypically matched controls. The compound action potential was distinguished from postsynaptic field potentials by its shorter latency, its persistence following application of cobalt or kynurenic acid that blocked postsynaptic responses, and its ability to follow paired-pulse stimulation at 300 Hz. Blockade of the postsynaptic field responses by kynurenic acid indicates that in the mouse, as in the rat, glutamate is the olfactory nerve transmitter. The mean conduction velocity of ORNs in wild-type control mice was 0. 47+/-0.19 (S.E.M.) m/s (n=5), similar to the conduction velocity reported for other mammals. The mean diameter of ORN axons in control mice was 0.202+/-0.005 and 0.261+/-0.006 microm in the OMP-null mice. This increase in fiber diameter in the OMP-nulls predicts an increase in impulse conduction velocity. However, the mean conduction velocity of OMP-null mice, 0.38+/-0.03 m/s (n=6), was not significantly different from control (P>0.1). The conduction velocity predicted by the increase in fiber diameter in OMP-null mice was within the 95% confidence interval of the measured value. Thus, OMP-null ORNs are normal with respect to the conduction velocity of their axons. The number of axodendritic synapses in the glomeruli of OMP-null mice is higher than in congenic wild-type mice. PMID- 10825499 TI - Biophysical properties and responses to glutamate receptor agonists of identified subpopulations of rat geniculate ganglion neurons. AB - The goal of the current study was to evaluate the electrophysiological properties and responses to glutamate receptor agonists of rat geniculate ganglion (GG) neurons innervating the tongue. Subpopulations of GG neurons were labeled by injecting Fluoro-Gold (FG) or True Blue chloride into the anterior tongue and soft palate (AT and SP neurons) and applying FG crystals to the posterior auricular branch of the facial nerve (PA neurons). Three to 12 days later, the GG neurons were acutely isolated and patch clamped. Although many biophysical properties of the AT, SP and PA neurons were similar, significant differences were found among these groups in properties related to cell excitability. For example, the average amount of current necessary to elicit an action potential was 61 pA in AT neurons (n=55), 90 pA in SP neurons (n=41) and 189 pA in PA neurons (n=35, P<0.001). In addition, AT neurons tended to fire significantly more action potentials during depolarization as well as following hyperpolarizing pulses than SP or PA neuron types. Most GG neurons responded to application of glutamate receptor agonists. The neurons responded with a depolarization accompanied by a reduction in input resistance. These results suggest that subpopulations of neurons in the geniculate ganglion have distinct biophysical properties and express functional glutamate receptors. The differing biophysical properties of GG neurons is possibly related to their functional heterogeneity and glutaminergic neurotransmission may function in the processing of gustatory, and other sensory information, within the geniculate ganglion and its projections. PMID- 10825500 TI - Pre- and postsynaptic actions of serotonin on rat suprachiasmatic nucleus neurons. AB - Serotoninergic transmission is implicated in the photic and non-photic regulation of circadian rhythms. 5-HT (1-100 microM), carboxamidotryptamine (5-CT 0.1-10 microM) and (+)-8-hydroxy-dipropylaminotetraline (8-OH-DPAT, 1-30 microM) dose dependently activated an outward current (5-100 pA) in 30% of neurons voltage clamped at -60 mV in the suprachiasmatic nucleus (SCN) in vitro slice. EC(50) values were 7.0 microM for 5-HT and 0.2 microM for 5-CT. Serotonin-induced outward current was associated with an increase in input conductance, and the current was blocked by Ba(2+) (1 mM). The amplitude of the current was enhanced by depolarization, reduced by hyperpolarization, and reversed its polarity during a hyperpolarization beyond the potassium equilibrium potential. Mean amplitudes of the 5-HT outward current changed with time of the subjective circadian day. The value near CT2 (23.8 pA) was about 4 times greater than that around CT14 (6.7 pA). Cells that responded with an outward current showed four types of morphology: monopolar, simple bipolar, curly bipolar and radial shaped; they were localized in all parts of the SCN. The EPSC evoked by retino-hypothalamic-tract (RHT) stimulation was inhibited 26% but the inward current induced by exogenously applied glutamate or NMDA was not affected by serotonin agonists. Focal stimulation-induced and spontaneous IPSC but not the exogenous GABA-induced outward current were inhibited by 5-HT agonists in a subpopulation of cells. In conclusion, 5-HT regulates SCN neurons by both pre- and post-synaptic inhibitory mechanisms; the latter may play a key role in modulating SCN circadian rhythm by activation of 5-HT receptors and opening of a potassium channel. PMID- 10825501 TI - Brain-derived neurotrophic factor in Huntington disease. AB - Trophic factors, administered systemically or delivered via genetically-modified cells grafted into target regions, have been proposed as putative therapeutic agents in human neurodegenerative disorders. In parallel to the study of the beneficial effects in experimental models of particular diseases, a crucial aspect of the study of trophic factors is the gathering of information about the actual trophic factor expression in human diseased states. Brain-derived neurotrophic factor (BDNF) promotes survival and growth of various nerve cell populations during normal development and following various insults in the developing and adult brain. In particular, BDNF prevents cell death of certain striatal populations in excitotoxic models of Huntington disease (HD) following intrastriatal injection of quinolinic acid to the adult rodent brain. The present study examines BDNF expression, by gel electrophoresis and Western blotting, and immunohistochemistry, in the brains of patients who had suffered from HD. Reduced BDNF expression, ranging from 53 to 82%, has been found in the caudate and putamen in HD when compared with age-matched controls. No modifications in BDNF expression levels have been seen in the parietal cortex, temporal cortex and hippocampus. Furthermore, immunohistochemistry has shown reduced BDNF immunoreactivity in caudate neurons, but not in cortical neurons in HD when compared with controls. These data demonstrate selective BDNF decay in regions that are vulnerable to HD, and suggest, in combination with results in experimental models, that a BDNF surplus may have beneficial effects in the treatment of HD. PMID- 10825502 TI - Circadian modulation of fos responses to odor of the red fox, a rodent predator, in the rat olfactory system. AB - We have previously shown that neuronal responses to a biologically neutral odor, cedar wood oil, in the olfactory system are greater in the subjective night compared to subjective day. In the present study, we confirm these results and extend them to a biologically relevant odor, the urine of the red fox, a rodent predator. Fos induced by exposure of rats to fox urine or a neutral odor, mineral oil, was markedly enhanced during the subjective night compared to subjective day in the main olfactory bulb, primary olfactory cortex, and other structures related to olfaction. These results show that neuronal responses to an ethologically relevant odor follow a circadian rhythm similar to biologically neutral odors. Fos responses induced by fox urine were observed to be of greater magnitude than a neutral odor in brain areas involved in fear responses, suggesting that fox urine activates fear circuitry. PMID- 10825503 TI - Developmental stage-dependent protective effect of NGF against lead cholinotoxicity in the rat septum. AB - The ability of nerve growth factor (NGF) to ameliorate developmental cholinotoxicity of inorganic lead (Pb) for the septal neurons was investigated by making intracerebroventricular injections of single doses of 30 microg 2.5S NGF into maternally lead-exposed suckling rats on postnatal days P2, P4, P11, or P18. Administration of NGF on P4 or later induced septal choline acetyltransferase (ChAT) activity to the same relative extent in both Pb-exposed as in control rats but failed to reverse the net reductions of ChAT activity induced by Pb. In contrast, injection of NGF at P2 completely restored ChAT activity in Pb-exposed pups to control levels by preventing the loss of ChAT-immunoreactive cells in the septum. It is concluded that although NGF retains the capacity to upregulate ChAT throughout the period of Pb exposure, it protects against the Pb-induced loss of septal cholinergic neurons only when applied within the critical period of Pb vulnerability between postnatal days 2 and 4. PMID- 10825504 TI - Immunohistochemical study on the distribution of the voltage-gated calcium channel alpha(1B) subunit in the mature rat brain. AB - Many neuronal processes are regulated by calcium influx through voltage-gated calcium channels. Among these calcium channels, the N-type channel has been shown to play a significant role in the regulation of neurotransmitter release. This present report has demonstrated the distribution of the alpha(1B) subunit of the N-type calcium channel in the mature rat brain. The most striking findings were the highest immunoreactivity in the CA1 region of the hippocampus and the clear demarcation in the thalamus. Characteristic patterns of distribution were noted in the cerebral cortex, hippocampus, caudate-putamen, thalamus, and cerebellum. In the cerebral cortex, strong staining was seen in layers II-III and V. In the basal ganglia, the staining intensity was high in the caudate-putamen, whereas there was no immunoreactivity in the globus pallidus. Although the staining intensity was relatively low compared to that of the forebrain, moderate alpha(1B)-IR was found in the dorsal cochlear nucleus and mesencephalic trigeminal nucleus. The detailed features of this study will extend our current knowledge of alpha(1B) subunit distribution in terms of the number of neuroanatomical areas examined, by investigating alpha(1B) subunit protein expression, for the first time. PMID- 10825505 TI - CNS voltage-dependent Na(+) channel expression and distribution in an undifferentiated and differentiated CNS cell line. AB - Upon serum removal, CAD-R1 cells undergo neurite outgrowth and an increase in voltage-dependent Na(+) current (VDNaC) density without changing their activation and inactivation properties. Insulin and endothelial cell growth supplement inhibited the increase in VDNaC density but not the neurite outgrowth. RI, RII, RIII Na(+) channel proteins were expressed in CAD-R1 cells. These proteins exhibited both similar and different distribution and clustering patterns which suggested the channel's structural differences play a role in channel distribution. PMID- 10825506 TI - The expression of transforming growth factor-beta1 (TGF-beta1) in hippocampal neurons: a temporary upregulated protein level after transient forebrain ischemia in the rat. AB - Exogenous TGF-beta1 has been shown to protect neurons from damage induced in vitro and in vivo. In this study we attempted to examine the expression of endogenous TGF-beta1 mRNA and protein in the hippocampus of non-ischemic and ischemic rats, and to localize TGF-beta1 protein and DNA fragmentation by double staining. Transient ischemia was induced for 10 min in Wistar rats by clamping both common carotid arteries and lowering blood pressure to 40 mmHg. Bioactive TGF-beta1 was selectively determined in CA1 pyramidal neurons of non-ischemic rats. It was upregulated after 3 h and 6 h of reperfusion corresponding to the increase in TGF-beta1 mRNA level detected by RT-PCR. Lectin and GFAP staining showed no detectable activated microglial cells and astrocytes in the hippocampus 3 h and 6 h after ischemia. When neuronal damage proceeded through day 2 to day 4 after ischemia as demonstrated by TUNEL-staining, TGF-beta1 immunoreactivity (ir) disappeared in damaged neurons but persisted in viable neurons although TGF-beta1 mRNA levels continuously increased. Double-staining revealed that TUNEL-positive neurons did not express TGF-beta1, while TUNEL-negative neurons in the CA1 subfield exhibited a distinct TGF-beta1 ir. These data indicate that hippocampal CA1 neurons can express TGF-beta1 under physiological conditions and upregulate its expression during the first hours after ischemia, that is independent of the activation of glial cells. The endogenous TGF-beta1 expressed in neurons may play a role in the pathological process of DNA degradation and delayed neuronal death after transient forebrain ischemia. PMID- 10825507 TI - mu-Calpain activation, DNA fragmentation, and synergistic effects of caspase and calpain inhibitors in protecting hippocampal neurons from ischemic damage. AB - The differentiated cells seem to share the ability to induce their own death by the activation of an internally encoded suicide program. When activated, this suicide program initiates a characteristic form of cell death called apoptosis. A central challenge in apoptosis research is understanding the mechanisms by which apoptotic cascades are initiated and affected. We tested a potential role for calpain in the programmed cell death under ischemic conditions and found that calpain is (1) activated at a time preceding morphological changes, DNA fragmentation and death, (2) that calpain is translocated to the nucleus before DNA laddering, (3) pretreatment with caspase inhibitors and/or calpain inhibitors block not only the proteolytic actions of the enzyme, but also the cell death process itself in the CA1 subfield after transient global ischemia in a synergistic manner. In conclusion, the present results contribute additional evidence that proteases may play a functional role in apoptotic cell death and extend them to include the possibility that endogenous proteases are capable of inducing the striking DNA fragmentation and chromatin condensation, which are the principle criteria currently used to define apoptotic death. Moreover, the synergistic effect of caspase and calpain inhibitors in protecting neurons form ischemic damage suggests that there is a cross-talk between caspase and calpain during apoptosis. PMID- 10825508 TI - Intracerebral hemodynamics probed by near infrared spectroscopy in the transition between wakefulness and sleep. AB - Previous imaging studies have shown that cerebral metabolism is gradually reduced at the beginning of sleep. Few studies have examined the sleep state transition periods from wakefulness to sleep and sleep to wakefulness. The current study used the Near Infrared Spectroscopy (NIRS) technique to describe the intracerebral hemodynamics at the frontal pole in the circumscribed period between wakefulness and sleep. Nine healthy young adults were studied during afternoon naps. Optical probes were placed on the forehead and EEG electrodes on the scalp. At sleep onset oxygenated hemoglobin (oxy-Hb) was reduced (P<0.01) and deoxygenated hemoglobin (deoxy-Hb) showed a near significant reduction (P<0.063). At sleep offset there were increases in oxy-Hb (P<0.005) and deoxy-Hb (P<0.05). In 18 of 26 transitions to sleep there was a coordinated fall in both NIRS parameters, we call the Switch Point, that lasted a mean of 3.6 s. In 32 of 36 transitions to wakefulness there was an analogous Switch Point that lasted a mean of 3.4 s. Before and after the Switch Point, changes were small and the relationship between oxy-Hb and deoxy-Hb was a combination of parallel and reciprocal fluctuations. A synchronized, parallel and short-lived change in oxy Hb and deoxy-Hb is a discrete event in the transition period between wakefulness and sleep. The concentration of these light absorbing molecules is abruptly set to a new level at sleep-wake transitions and probably reflects the different perfusion demands of these states. PMID- 10825509 TI - Overexpression of neurofilament subunit M accelerates axonal transport of neurofilaments. AB - Neurofilaments are composed of three polypeptide subunits (NF-H, NF-M and NF-L). They are the most abundant cytoskeletal element in large myelinated axons and play a central role in development of axonal caliber. To perform this role, neurofilaments are transported from their site of synthesis, the cell bodies, to the distal axons. Previous studies showed that overexpression of NF-M in transgenic mice led to accumulation of neurofilaments in neurons and a reduction in the number of neurofilaments in axons, suggesting that axonal transport of neurofilaments was slowed. To determine whether this was the case, we measured axonal transport velocities in the wild type and transgenic mice overexpressing NF-M by the classical pulse-labeling method using 35S-methionine. We found that neurofilament transport in peripheral motor axons can be described with a model consistent with two linear velocities. Contrary to expectations, both velocities were accelerated by overexpression of NF-M. These results suggest that subunit composition in neurofilaments play a regulatory role in neurofilament transport. In addition, these results show that there are regional differences in neurofilament transport along long axons and these differences may be the basis for selective regional accumulation of neurofilaments in various neurological disorders. PMID- 10825510 TI - In this issue ellipsis PMID- 10825511 TI - Achievements of the last century in neurosurgery and a view to the 21st century. PMID- 10825512 TI - Delayed magnetic resonance imaging with GdD-DTPA differentiates subdural hygroma and subdural effusion. AB - BACKGROUND: Posttraumatic subdural fluid collection is not a single clinical entity but can be divided into subdural hygroma and subdural effusion. Appropriate treatment requires preoperative differentiation. Delayed magnetic resonance (MR) imaging with intravenous administration of gadolinium diethylenetriaminepenta-acetic acid (Gd-DTPA) was used to differentiate subdural hygroma and subdural effusion. METHODS: Timed arterial blood specimens were taken after intravenous Gd administration in patients with posttraumatic subdural fluid collections (five subdural hygromas and 13 subdural effusions). Delayed MR imaging was performed 1 hour after administration of Gd-DTPA. Gd-DTPA concentrations in the subdural fluid and blood specimens were measured by ion coupled plasma emission spectrometry. Dynamic biologic modeling was used to calculate the transfer rate constant for Gd-DTPA influx into these subdural fluid collections. RESULTS: The Gd concentrations in subdural hygromas and subdural effusions were 16 +/- 6 and 79 +/- 12 nmol/mL, respectively. The transfer rate constants for subdural hygromas and subdural effusions were 4.8 +/- 2.1 and 20.6 +/- 2.1 (x10(-4))min(-1), respectively. These values were significantly higher in subdural effusions than in subdural hygromas (p < 0.01). Delayed MR imaging with Gd showed significantly higher mean enhancement of 77.1 +/- 14.2% for subdural effusions compared to 4.6 +/- 3.1% for subdural hygromas (p < 0.01). CONCLUSIONS: Delayed MR imaging with Gd can differentiate subdural hygroma and subdural effusion. PMID- 10825513 TI - Spontaneous resolution of chronic subdural hematoma. AB - BACKGROUND: Spontaneous resolution of chronic subdural hematoma has rarely been reported in the literature, and its mechanism has not been fully investigated. Response to surgery has been very satisfactory; in fact, this is generally considered the treatment of choice. METHODS: From a series of 24 cases of chronic subdural hematomas, we observed five patients between 1996 to 1998. These patients showed headache and decrease of cognitive level, 4-5 weeks after minor head injury. Neurologic evaluation revealed only worsening of mental function according to Mini Mental State Examination (MMSE). Computed tomography (CT) scans showed brain atrophy and chronic subdural hematoma without increased intracranial pressure. These patients were treated by clinical observation and serial cerebral CT scans. RESULTS: After 7 to 10 days, all patients showed improvement of clinical signs. After 30 to 45 days, radiological disappearance or marked reduction in size of the hematoma and complete clinical recovery were obtained. No neurological deficits and no recurrences have been observed during follow-up (3 months to 2 years). CONCLUSIONS: We believe that age greater than 70 years, decreased cognitive level (MMSE = 21), brain atrophy, and absence of increase of intracranial pressure are clinical and radiological signs that allow one to choose conservative treatment. PMID- 10825514 TI - Combined neuroendovascular stenting and coil embolization for cervical carotid artery dissection causing symptomatic mass effect. AB - BACKGROUND: Carotid artery dissection manifesting with symptomatic mass effect has been treated surgically according to the previous literature. Recently, some cases of carotid artery dissection manifesting with ischemic symptoms were treated successfully with endovascular insertion of coils after stenting. METHODS: A 42-year-old man with spontaneous dissection of the left cervical internal carotid artery (ICA) presented with the major complaint of left neck swelling and pain that was considered to be the mass effect of a pseudoaneurysm caused by dissection of the ICA. Endovascular therapy using a stent and coils was performed. The self-expanding stent was deployed to cover the neck of the pseudoaneurysm. A microcatheter was then guided through the stent mesh into the aneurysm, and coils were placed to pack it. RESULTS: Four months later, angiography revealed complete embolization of the aneurysm with preserved flow in the ICA. The mass effect attributable to the pseudoaneurysm was relieved symptomatically as well as radiologically. CONCLUSION: Cervical artery dissection with symptomatic mass effect++ can be treated successfully by the combination of stent and coils. This may be considered as an alternative to conventional proximal ligation, extracranial-intracranial bypass, or direct surgical repair. PMID- 10825515 TI - Effect of an antioxidant, ebselen, on development of chronic cerebral vasospasm after subarachnoid hemorrhage in primates. AB - BACKGROUND: Oxidation and/or free radical reactions after subarachnoid hemorrhage (SAH) may be involved in the development of chronic cerebral vasospasm. The inhibition of these reactions is thought to be one of the therapeutic strategies for prevention of cerebral vasospasm. We investigated the effect of Ebselen, a synthetic seleno-organic compound, which exhibits anti-oxidation by glutathione peroxidaselike activity to inhibit free radical reactions by lipid peroxidation on the development of chronic cerebral vasospasm in a primate model. METHODS: Seventeen monkeys were used. SAH was produced by introduction of a blood clot around the right middle cerebral artery and the right side of the circle of Willis in all animals. The monkeys were randomly divided into three groups according to Ebselen dosage: 1) no dosage or non-treated group; 2) high-dose Ebselen group; and 3) low-dose Ebselen group. The drug was administered at 10 mg/Kg in the high-dose group and 5 mg/Kg in the low-dose group twice a day in each group for 7 days after SAH. The vessel diameter was evaluated on angiograms before the induction of SAH and at Day 7 following SAH. RESULTS: In the untreated group, the angiograms showed significant (p < 0.05) reductions of the mean vessel caliber of the right internal carotid (ICA) (38 +/- 10% reduction) and the middle cerebral artery (MCA) (56 +/- 9.7%) compared with the baseline value before SAH. In the high-dose Ebselen-treated group, the mean percent reduction in vessel caliber of the right ICA (16 +/- 11%) and MCA (28 +/- 9.5%) on Day 7 angiograms were significantly (p < 0.05) lower than those in the nontreated group, whereas the mean percent reduction of these vessels in the low-dose Ebselen-treated group showed no significant difference compared with the untreated group. CONCLUSIONS: Chronic cerebral vasospasm was inhibited in the animals in which a relatively large amount of Ebselen was administered for 7 days after SAH. The results suggest that the oxidation or free radical reaction by lipid peroxidation after SAH might be involved in the pathogenesis of vasospasm, and that inhibition of these reactions by drugs, such as Ebselen, may have a promising effect for prevention of vasospasm. PMID- 10825516 TI - A combination of wrapping and clipping using a collagen-impregnated dacron fabric (Hemashield). AB - BACKGROUND: We describe techniques combining wrapping and clipping using a collagen-impregnated Dacron knitted fabric (Hemashield) for accidental arterial perforations and broad-based aneurysms. The results of these techniques in seven patients are presented. METHODS: Clip-reinforced wrapping was performed to obtain hemostasis in two patients with arterial perforations and in a patient with a ruptured broad-based aneurysm in the internal carotid artery. Clipping of the broad neck of the aneurysm and wrapping with Hemashield (wrap-clipping) was performed in four patients with unruptured aneurysms (one internal carotid artery, two middle cerebral artery, one basilar artery). RESULTS: In the three patients treated with clip-reinforced wrapping, complete hemostasis was obtained just after clip application. In the patient with a ruptured broad-based aneurysm, postoperative angiography demonstrated that the dome of the aneurysm was well compressed. In the four patients treated with wrap-clipping, postoperative angiography revealed successful clipping of the broad neck of the aneurysm. CONCLUSION: In this early experience, there were no problems in the use of Hemashield for clip-reinforced wrapping or wrap-clipping. PMID- 10825517 TI - Dissecting aneurysm of the anterior choroidal artery: angiographical and MR imaging findings. AB - BACKGROUND: Intracranial dissecting aneurysms are relatively rare. We present a rare case of a dissecting aneurysm originating from the anterior choroidal artery; this is the first reported case. CASE DESCRIPTION: A 42-year-old man suffered sudden onset of right hemiparesis and dysarthria. Computed tomography on admission revealed a small low density area in the posterior limb of the internal capsule. MR imaging revealed aneurysmal dilatation of the anterior choroidal artery, and angiography revealed aneurysmal dilatation and stasis of dye in the venous phase at the anterior choroidal artery, which resolved in the chronic stage. CONCLUSION: We describe a rare case of a dissecting aneurysm of the anterior choroidal artery. The radiological findings were characteristic of dissecting aneurysms in spite of the rare location. PMID- 10825518 TI - Motor area cavernous angioma: case report. PMID- 10825519 TI - Cervical discectomy. A prospective analysis of three operative techniques. AB - BACKGROUND: Cervical disc herniation causing neurological compromise is a common affliction. Sophisticated surgical treatments have been developed throughout the twentieth century and are largely successful. Although each procedure has its supporters, it is still unclear if one surgical technique is superior. METHODS: A prospective trial was designed to evaluate the efficacy of three surgical procedures for the treatment of cervical radiculopathy caused by a unilateral acute herniated cervical disc. Patients were randomized to posterior cervical foraminotomy (FOR), and anterior cervical discectomy with (ACDF), and without (ACD) fusion. Perioperative data, office follow-up and long-term follow-up were used to compare the procedures. RESULTS: All of the procedures yielded excellent relief of symptoms and signs postoperatively and during follow-up. Operative time and hospital stay were slightly shorter for ACD compared with ACDF and FOR. Reoperations occurred in all groups but there was a trend for higher recurrence at the same level with FOR and recurrence at other levels with ACDF. CONCLUSION: All three of the procedures were successful for treatment of cervical radiculopathy caused by a herniated cervical disc. Although the numbers in this study were small, none of the procedures could be considered superior to the others. This study suggests that the selection of surgical procedure may reasonably be based on the preference of the surgeon and tailored to the individual patient. PMID- 10825520 TI - Anatomic considerations of costotransverse screw placement in the thoracic spine. AB - BACKGROUND: Numerous techniques have been reported to restore spinal stability and to correct spinal deformities, including rods with wires/hooks, and rods or plates with pedicular screws. It was thought that posterior fixation of the thoracic spine through the costotransverse joint may be another alternative. METHODS: Nine cadavers were obtained for study of screw fixation of the costotransverse joint for posterior thoracic instrumentation. The entrance point for screw insertion was designed to be at the posterior center of the clubbed extremity of the transverse process. From this point, a 3-mm drill bit was used to create the screw path penetrating the costotransverse joint and the ventral cortex of the rib. Under direct visualization of the costotransverse joint, the drill bit was directed parallel to the sagittal plane and toward the upper portion of the rib. Measurements included the screw path length and sagittal angulation. Also, the distance between the superior borders of the transverse process and the tubercle of the rib and the anatomic relationship of the drill bit exit to the intercostal vessels and nerves were evaluated. RESULTS: The maximum length of the screw path was found at T1 (19.7 mm), whereas the minimum length was noted at T4-T5 (13.9 mm). This value decreased gradually from T1 to T4 T5, and slightly increased to T10. The larger sagittal angles of the screw path were found at the levels of T1-T4 (78-86 degrees ), whereas the smaller were noted at the levels below T5 (53-61 degrees ). The mean distance between the superior borders of the transverse process and the tubercle of the rib was smaller at T1-T5 (0.2-0.4 mm), and significantly increased to T8 (5. 1 mm), and then slightly decreased to T10. The variation of this parameter was remarkable. All of the exit points for the drill bit were located in the upper half of the rib, and away from the intercostal vessels. CONCLUSIONS: The ideal screw orientation is parallel to the sagittal plane, and angled 80-90 degrees relative to the frontal plane for T1-T4 and 50-70 degrees for T5-T10 superiorly, starting at the posterior center of the transverse process. Costotransverse screw fixation in the thoracic spine may be an alternative to pre-existing methods. PMID- 10825521 TI - Spontaneous brachial plexus hemorrhage-case report. AB - BACKGROUND: Shoulder hemorrhage resulting in brachial plexus neuropathy is a rare occurrence most often seen in cases of traumatic injury or anticoagulation therapy. We report a unique case of spontaneous brachial plexus hemorrhage. CASE DESCRIPTION: This is the first report of a spontaneous shoulder hemorrhage in which a 48-year-old jackhammer operator presented to the emergency department with a sudden onset of right shoulder pain and upper extremity pain and numbness. Imaging studies revealed a hematoma in the right axilla and chest wall. Without evidence of active bleeding or worsening neurologic deficit, this patient was treated conservatively with pain control and observation and eventually experienced a full recovery. Had there been persistent neurologic deficit, however, surgical evacuation would have been indicated. CONCLUSIONS: Cases of nerve compression caused by a hematoma should be analyzed on the basis of the severity of the neurologic deficit and not on the underlying cause of bleeding. Conservative treatment may be indicated in cases of mild or improving neurologic deficit, but regardless of its etiology, a hematoma that results in severe or worsening neurologic symptoms must be surgically evacuated to prevent permanent nerve damage. PMID- 10825522 TI - Antisense-mediated inhibition of the bcl-2 gene induces apoptosis in human malignant glioma. AB - BACKGROUND: The bcl-2 protooncogene represses a number of cellular apoptotic pathways and is known to be expressed in increasing amounts in glial tumors of higher malignancy. We tested whether antisense oligonucleotides to the bcl-2 gene would affect glioma cell viability. METHODS: Antisense oligonucleotides directed to the first six codons of the human bcl-2 gene, and nonsense oligonucleotides as a control, were transfected into malignant glioma cells. Two human Bcl-2 positive glioblastoma cell lines from our tumor bank (Jon52 and Roc) were both transfected in vitro with bcl-2 antisense (AS) and nonsense (NS) oligonucleotides at 1 microm and 5 microm concentrations for 5 and 24 hr. Cell viability was assessed at 2, 4, 5, and 7 days by using an MTT mitogenic assay and by cell counting via direct visualization using a hemocytometer. RESULTS: There was up to a log-fold decrease in cell growth of the bcl-2 AS treated cells compared to the NS transfected cells for both Roc (p = 0.007 and p = 0.004) and Jon52 (p = 0.02 and p = 0.004) at 5 and 24 hr of transfection. There was as much as 50% cytotoxicity in both glioblastoma cell lines at 1 microm and 5 microm concentrations after 24 hr transfection with AS bcl-2 oligonucleotides (all p < 0.01). Western blot analysis demonstrated a decrease in the expression of the Bcl-2 protein in one cell line, whereas there was a statistically significant increase in the apoptotic index of both cell lines (p < 0.05 by chi square analysis). CONCLUSIONS: Our results suggest that transfection of human glioma cells with antisense bcl-2 results in an increase in apoptotic death. This provides evidence that Bcl-2 plays a role in tumor progression of glioma by acting as an oncogene, and suggests that inhibition of the bcl-2 gene could have a therapeutic effect. PMID- 10825523 TI - Current surgical results of retrosigmoid approach in extralarge vestibular schwannomas. AB - BACKGROUND: Vestibular schwannomas (VS) are common tumors that can be cured; they are reported to comprise 6 approximately 8% of all intracranial tumors. The primary objective in the surgery of extralarge VS is total removal of the tumor mass while preserving the facial nerve. In extralarge tumors, complete excision of the tumor carries a significant risk of injuring the facial nerve and adjacent vital brain structures. The authors are reporting the techniques and results of operation on extralarge VS. METHODS: The material consisted of 30 patients during the last 6 years with surgically treated VS that had a maximal extrameatal diameter exceeding 4 cm. Suboccipital craniotomy and tumor removal was performed with patients in the lateral position. Results and complications of the surgical technique will be reviewed. RESULTS: Average age of patients was 45.2 years; there was a slight female predominance (1.5:1). Size of the mass ranged from 41 to 70 mm; all were removed by the retrosigmoid transmeatal approach. Peritumoral edema on MRI was seen in 50% (15/30). Total removal was achieved in 73.3% (22/30) with no significant relationship to peritumoral edema. In the cases of total removal, the facial nerve preservation rate was 86.4% (19/22). There was no mortality. Surgical complications were hemorrhage and CSF leakage in 1 case (3%) and 8 cases (26.7%), respectively, but in most of these cases, conservative treatment was adequate. In patients in whom anatomic preservation of the facial nerve was achieved, facial nerve function improved progressively within a year. In all cases except for one with gait disturbance, a good outcome was achieved. CONCLUSIONS: Our surgical techniques, including the prediction of facial nerve displacement, not using retractors, and replacement of bone, contributed to good surgical results in a series of extralarge VS. PMID- 10825524 TI - Skull metastasis of hepatocellular carcinoma associated with acute epidural hematoma: a case report. AB - BACKGROUND: Although epidural hematoma is well documented in trauma patients, its association with other etiologies, such as neoplasms, is not widely known. Here the authors report a case of acute epidural hematoma that originated from a metastatic hepatocellular carcinoma (HCC) in the skull. CASE DESCRIPTION: A 70 year-old male was admitted to our hospital with left-sided hemiparesis. Preoperative computed tomography (CT) revealed a lenticular high-density area adjacent to the right parietal bone, consistent with an acute epidural hematoma. A subsequent magnetic resonance image (MRI) showed a skull tumor adjacent to the epidural hematoma. Removal of the tumor and evacuation of the hematoma were performed and the pathological diagnosis was metastatic HCC. Postoperatively, the patient gradually recovered but he died of pneumonia 2 months later. CONCLUSION: This report represents an additional example of a rare case of metastatic skull tumor associated with acute epidural hematoma. The authors suggest that metastatic skull tumors may be one of the important differential diagnoses in patients with acute epidural hematoma. PMID- 10825525 TI - Radiosurgery for bilateral neurinomas associated with neurofibromatosis type 2. AB - OBJECTIVE: The clinical course of bilateral acoustic tumors associated with neurofibromatosis (NF2) is generally troublesome, and no definite treatment strategy has been established. Follow-up results of bilateral acoustic tumors after radiosurgery are reported herein. METHODS: The current indications for radiosurgery are 1) a growing tumor less than 30 mm in mean diameter, 2) the ipsilateral ear has no serviceable hearing, and 3) there is risk of brain stem compression or brain stem dysfunction. Twenty cases of bilateral acoustic tumors were treated with the gamma knife, including 7 males and 13 females. The mean age was 38.2 years and the mean tumor size 24.4 mm. The tumors were treated with mean maximum and marginal doses of 26.8 Gy and 13.0 Gy, respectively. Among them, 12 patients had profound hearing loss in the ipsilateral (treated) ear, but the other 8 had serviceable hearing. RESULTS: Tumors treated with radiosurgery showed central necrosis in 60% of the cases at 6 months and in 70% at 9 months after radiosurgery. Thereafter, the tumors often demonstrated slow regression. The rate of tumor shrinkage was 20% at 12 months, 35% at 24 months, and almost 60% at 36 months. At the last follow-up (mean 33.6 months), the tumors demonstrated shrinkage in 50% and tumor control in 100%. The contralateral tumors were stable in 12 (60%) and enlarged in 8 (40%). Preservation of serviceable hearing ipsilaterally was obtained in 33.3%. Deterioration of ipsilateral facial nerve function, either in the natural course or as a complication, occurred in 10%. CONCLUSIONS: Because of good tumor control and tumor shrinkage as well as an acceptable complication rate, radiosurgery should be incorporated in the treatment strategy for bilateral acoustic tumors associated with NF2. PMID- 10825526 TI - Bone flap fixation with titanium clamps: a new technique. AB - BACKGROUND: Craniotomy bone flaps are replaced for both cosmetic and protective purposes. The purpose of this study was to determine the facility, strength, timing, and cost effectiveness of an alternative system for securing bone flaps. The system consists of titanium discs that clamp the bone flap to the skull edge. METHODS AND RESULTS: Ten cadaver craniotomy flaps each were reattached with either 24-gauge wire, miniplates, or titanium clamps. The titanium clamp system required significantly less time to fix than either wire or miniplates. The clamps were stronger than wire and cost less than miniplates. CONCLUSION: The titanium clamp system is a reasonable alternative to present craniotomy fixation methods with respect to ease of use, time consumption, strength, and relative cost. PMID- 10825527 TI - "Spontaneous" reduction of tonsillar herniation in acromegaly: a case report. PMID- 10825528 TI - Physicians and unions: an oxymoron? PMID- 10825529 TI - Structure, specificity and function of cyclomaltodextrinase, a multispecific enzyme of the alpha-amylase family. AB - Cyclomaltodextrinase (CDase, EC 3.2.1.54), maltogenic amylase (EC 3. 2.1.133), and neopullulanase (EC 3.2.1.135) are reported to be capable of hydrolyzing all or two of the following three types of substrates: cyclomaltodextrins (CDs); pullulan; and starch. These enzymes hydrolyze CDs and starch to maltose and pullulan to panose by cleavage of alpha-1,4 glycosidic bonds whereas alpha amylases essentially lack activity on CDs and pullulan. They also catalyze transglycosylation of oligosaccharides to the C3-, C4- or C6-hydroxyl groups of various acceptor sugar molecules. The present review surveys the biochemical, enzymatic, and structural properties of three types of such enzymes as defined based on the substrate specificity toward the CDs: type I, cyclomaltodextrinase and maltogenic amylase that hydrolyze CDs much faster than pullulan and starch; type II, Thermoactinomyces vulgaris amylase II (TVA II) that hydrolyzes CDs much less efficiently than pullulan; and type III, neopullulanase that hydrolyzes pullulan efficiently, but remains to be reported to hydrolyze CDs. These three types of enzymes exhibit 40-60% amino acid sequence identity. They occur in the cytoplasm of bacteria and have molecular masses from 62 to 90 kDa which are slightly larger than those of most alpha-amylases. Multiple amino acid sequence alignment and crystal structures of maltogenic amylase and TVA II reveal the presence of an N-terminal extension of approximately 130 residues not found in alpha-amylases. This unique N-terminal domain as seen in the crystal structures apparently contributes to the active site structure leading to the distinct substrate specificity through a dimer formation. In aqueous solution, most of these enzymes show a monomer-dimer equilibrium. The present review discusses the multiple specificity in the light of the oligomerization and the molecular structures arriving at a clarified enzyme classification. Finally, a physiological role of the enzymes is proposed. PMID- 10825530 TI - Isolation, characterization and binding properties of two rat liver fatty acid binding protein isoforms. AB - Mammalian liver has only one fatty acid-binding protein (L-FABP) while the liver of non-mammalian vertebrates expresses a liver basic FABP (Lb-FABP) in addition to other members of the FABP family. We explore the possibility that L-FABP isoforms accomplish, in the liver of mammals, the metabolic functions corresponding to the different FABPs present in the liver of non-mammalian vertebrates. We have isolated isoforms I and II which have a different residue 105, Asn in the former and Asp in the latter. We made a conformational comparison of the apo-isoforms by intrinsic fluorescence emission and fourth-derivative spectroscopy, native-state proteolysis and unfolding curves. Ligand affinity was studied by measuring cis-parinaric acid displacement by different ligands. They have differences in their molecular conformation, including the environment of the binding site. Isoform II has probably a more open conformation than isoform I, thus allowing the binding of a greater variety of ligands. The affinity of isoform II for lysophospholipids, prostaglandins, retinoids, bilirubin and bile salts is greater than that of isoform I. These characteristics of rat L-FABP isoforms I and II suggest that they may accomplish different functions as happens with those of the different FABP types in non-mammalian species. PMID- 10825531 TI - Overproduction in Escherichia coli, purification and characterization of a family I.3 lipase from Pseudomonas sp. MIS38. AB - Determination of the nucleotide sequence of the gene encoding a lipase from Pseudomonas sp. MIS38 (PML) revealed that PML is a member of the lipase family I.3 and is composed of 617 amino acid residues with a calculated molecular weight of 64510. Recombinant PML (rPML) was overproduced in Escherichia coli in an insoluble form, solubilized in the presence of 8 M urea, purified in a urea denatured form and refolded by removing urea in the presence of the Ca(2+) ion. Gel filtration chromatography suggests that this refolded protein is monomeric. rPML showed relatively broad substrate specificities and hydrolyzed glyceryl tributyrate and olive oil with comparable efficiencies. rPML was active only in the form of a holo-enzyme, in which at least 12 Ca(2+) ions bound. These Ca(2+) ions bound too tightly to be removed from the protein upon dialysis, but were removed from it upon EDTA treatment. The resultant apo-enzyme was fully active in the presence of 10 mM CaCl(2), but was inactive in the absence of the Ca(2+) ion. PML has a GXSXG motif, which is conserved in lipases/esterases and generally contains the active-site serine. The mutation of Ser(207) within this motif to Ala completely inactivated PML, suggesting that Ser(207) is the active-site serine of PML. PMID- 10825532 TI - Salvage pathway for NAD biosynthesis in Brevibacterium ammoniagenes: regulatory properties of triphosphate-dependent nicotinate phosphoribosyltransferase. AB - As the rate-limiting enzyme, catalyzing the first reaction in NAD salvage synthesis, nicotinate phosphoribosyltransferase (NAPRTase, EC 2.4.2.11) is of important interest for studies of intracellular pyridine nucleotide pool regulation. We have purified NAPRTase 520-fold from Brevibacterium ammoniagenes ATCC 6872 without using an over-expression system by applying acid treatment, salt fractionation, Ca-phosphate gel treatment, anion exchange column chromatography and size-exclusion gel filtration. Unlike this enzyme from other sources, B. ammoniagenes NAPRTase was found to be controlled by the feedback inhibition by the end product NAD with K(i)=0.7+/-0.1 mM. The reaction products, pyrophosphate and nicotinate mononucleotide, also decreased the enzyme activity, as did other intermediates of NAD synthesis, such as AMP, ADP and a NAD direct precursor, nicotinate adenine dinucleotide or deamido NAD. The enzyme was observed to require a nucleoside triphosphate for its activity and showed the maximum affinity for ATP. The specificity, however, turned out to be poor, and ATP could be substituted by other nucleoside triphosphates as well as by sodium triphosphate. The kinetic characteristics of the enzyme are reported. For the first time, our data have experimentally revealed such complicated stimulatory and inhibitory effects by the intermediates of NAD biosynthesis on one of its salvage enzymes, NAPRTase. On the basis of these data, the key role of NAPRTase is discussed in light of the regulation of NAD metabolism in B. ammoniagenes. PMID- 10825533 TI - Aggregation, dissociation and unfolding of glucose dehydrogenase during urea denaturation. AB - The effect of urea on glucose dehydrogenase from Bacillus megaterium has been studied by following changes in enzymatic activity, conformation and state of aggregation. It was found that the denaturation process involves several transitions. At very low urea concentrations (below 0.5 M), where the enzyme is fully active and tetrameric, there is a conformational change as monitored by an increase in intensity of the tryptophan fluorescence and a maximum exposure of organized hydrophobic surfaces as reported by the fluorescence of 4,4'-dianilino 1,1'-binaphthyl-5.5'-disulfonic acid. At slightly higher urea concentrations (0.75-2 M), a major conformational transition occurs, as monitored by circular dichroism and fluorescence measurements, in which the enzyme activity is completely lost and is concomitant with the formation of interacting intermediates that lead to a highly aggregated state. Increasing urea concentrations cause a complete dissociation to lead first a partially and eventually the complete unfolded monomer. These phenomena are fully reversible by dilution of denaturant. It is concluded that after urea denaturation, the folding/assembly pathway of glucose dehydrogenase occurs with the formation of intermediate species in which transient higher aggregates appear to be involved. PMID- 10825534 TI - Structure at 1.44 A resolution of an N-terminally truncated form of the rat serum complement C3d fragment. AB - Complement component C3 plays a key role in the complement-mediated immune defence, and occupies a central position within the complement cascade system. One of its degradation products, C3dg, was purified from rat serum and crystallised in two different crystal forms as N-terminally truncated fragment. Despite the truncation and the lack of a significant portion of the N-terminus as compared to C3d, the structure of the fragment is highly similar to that of recombinant human C3d (Nagar et al., Science 280 (1998) 1277-1281). Structural details of the reactive site have been obtained, suggesting a possible mode of thioester bond formation between Cys-1010 and Gln-1013 and thioester bond cleavage in the transacylation reaction involving His-1126. The truncation at the N-terminus of C3d leads to the exposure of a surface of the molecule that favours dimerisation, so that in both crystal forms, the fragment is present as a dimer, with monomers related by a two-fold axis. PMID- 10825535 TI - Characterization of diphosphonucleotide phosphatase/phosphodiesterase from yellow lupin (Lupinus luteus) seeds. AB - A phosphatase cleaving the pyrophosphate bond in diphosphonucleotides and phosphodiester bond in various phosphodiesters (pH optimum at 6.25) was purified from yellow lupin (Lupinus luteus L.) seeds. The enzyme is 75 kDa monomeric glycoprotein (pI=6.4) with 4.4% of carbohydrate (mannose, N-acetylglucosamine, fucose and xylose). Analysis of its partial amino acid sequence (8 peptides, 101 amino acid residues) together with no divalent cation requirements for catalysis points out that the purified enzyme is different from known plant pyrophosphate cleaving enzymes (apyrases and inorganic pyrophosphatases). Its physiological role could be related to a regulation of diphosphonucleotides level in plant metabolism. PMID- 10825536 TI - Dissociation of mitochondrial malate dehydrogenase into active soluble subunits. AB - Gel exclusion chromatographic studies demonstrate that cytosolic and mitochondrial malate dehydrogenases (cMDH and mMDH) dissociate into subunits in the presence of 0.1% of the non-ionic detergent Triton X-100 (TX-100). The presence of cofactor and catalytically competent cofactor-substrate pairs does not protect mMDH against this dissociation. In contrast, cMDH dimers resist dissociation in the presence of either addition. Since steady state kinetic studies indicate both enzymes are fully active in the presence of 0.1% TX-100, we conclude that quaternary structure is not a kinetically important feature of mMDH structure and cooperativity does not account for mMDH kinetic anomalies. In contrast, cooperativity is a reasonable explanation for cMDH kinetic properties even in the presence of 0.1% TX-100, since this enzyme's subunits associate in the presence of active site ligands. The existence of fully active mMDH subunits raises the possibility that this species rather than the dimer may be a constituent of proposed multi-enzyme complexes of the mitochondrion. Preliminary chromatographic experiments involving gently disrupted mitochondria have found MDH activity in differently sized complexes, all with molecular weights larger than the mMDH dimer but smaller than complexes anticipated for multi-enzyme complexes involving other enzymes and the mMDH dimer. PMID- 10825537 TI - Effect of ligand binding on the flexibility of dihydrofolate reductase as revealed by compressibility. AB - The partial specific volume, v, and adiabatic compressibility, beta(s), of Escherichia coli dihydrofolate reductase were measured at 30 degrees C in the presence of various ligands (folate, dihydrofolate, tetrahydrofolate, NADPH, NADP, methotrexate, and KCl). Binding of these ligands (binary and ternary complexes) brought about large changes of v (0.734-0.754 cm(3) g(-1)) and beta(s) (6. 6x10(-6)-9.8x10(-6) bar(-1)), keeping a linear relationship between the two parameters. The values of v and beta(s) increased with an increase in internal cavity, V(cav), and a decrease in accessible surface area, ASA, which were calculated from the X-ray crystal structures of the complexes. A large variation of V(cav) relative to ASA by ligand binding suggested that the cavity is a dominant factor and the effect of hydration might be small for the ligand-induced changes of v and beta(s). The beta(s) values of the binary and ternary complexes suggested a characteristic conformational flexibility of the kinetic intermediates in the enzyme reaction coordinate. Comparison of beta(s) with the cavity distribution in the crystal structures revealed that the flexibility of the intermediates was mainly determined by the total cavity volume with minor contributions of the number, position, and size of cavities. These results demonstrate that the compressibility is a useful measure of the conformational flexibility of the intermediates in the enzyme reaction and that the combined study of compressibility and X-ray crystallography gives new insight into the protein dynamics through the behavior of the cavities. PMID- 10825538 TI - Why has porcine VEG protein unusually high stability and suppressed binding ability? AB - Von Ebner gland protein (VEGP) and odorant-binding protein (OBP) were purified from porcine lingual epithelium and nasal mucosa, respectively. Both VEGP and OBP preparations were homogeneous as indicated by SDS-PAGE, isoelectric focusing, gel filtration and electrospray mass spectrometry. However, high-sensitivity differential scanning calorimetry (HS-DSC) yielded multiphasic denaturation thermograms for both proteins indicating their conformational heterogeneity. The unfolding transition of VEGP is observed at extremely high temperatures (about 110 degrees C), which is unexpected for a protein with significant structural homology to OBP and other lipocalins. Isothermal titration calorimetry (ITC) did not detect the binding of either aspartame or denatonium saccharide to VEGP nor did it detect binding of 2-isobutyl-3-methoxypyrazine (IBMP) to OBP. Extraction of OBP with mixed organic solvents eliminated the conformational heterogeneity and the protein showed a reversible two-state transition in HS-DSC thereafter. ITC also showed that the extracted OBP was able to bind IBMP. These results imply that tightly bound endogenous ligands increase the thermal stability of OBP and block the binding of other ligands. In contrast to OBP, the extraction of VEGP with organic solvents failed to promote binding or to establish thermal homogeneity, most likely because of the irreversible denaturation of VEGP. Thus, the elucidation of the functional behaviour of VEGP is closely related to the exhaustive purging of its endogenous ligands which otherwise very efficiently mask ligand binding sites of this protein. PMID- 10825539 TI - LRDD, a novel leucine rich repeat and death domain containing protein. AB - Death domains (DD) and leucine rich repeats (LRR) are two different types of protein interaction motifs. Death domains are found predominantly in proteins involved in signaling and are involved in homo- and heteromultimerization. Leucine rich repeats are found in proteins with diverse cellular functions, like cell adhesion and cellular signaling, and mediate reversible protein-protein interactions. In this paper we report the cloning of a new human gene called LRDD (leucine repeat death domain containing protein). LRDD encodes a protein of 83 kDa with six LRRs at the N-terminus and a DD at the C-terminus. LRDD appears to be processed into two fragments of about 33 and 55 kDa, containing LRRs and DD respectively. Interestingly, LRDD is shown to interact with two other death domain containing proteins, FADD and MADD, presumably through death domain interactions. LRDD may represent a new type of adapter protein that could be involved in signaling or other cellular functions. PMID- 10825540 TI - Dissecting the nucleotide binding properties of Escherichia coli 6-hydroxymethyl 7,8-dihydropterin pyrophosphokinase with fluorescent 3'(2)'-o anthraniloyladenosine 5'-triphosphate. AB - 6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase (HPPK) catalyzes the transfer of pyrophosphate from ATP to 6-hydroxymethyl-7, 8-dihydropterin, the first reaction in the folate biosynthetic pathway. Like other enzymes in the folate pathway, HPPK is an ideal target for development of antimicrobial agents because the enzyme is essential for microorganisms but is absent from humans and animals. Using 3'(2')-o-anthraniloyladenosine 5'-triphosphate as a fluorescent probe, a fluorometric competitive binding assay has been developed for measuring the dissociation constants of various compounds that bind to the ATP site of HPPK. The fluorometric assay has been used to determine the nucleotide specificity and dissect the energetics of the binding of MgATP. The order of affinity of various nucleoside triphosphates for HPPK is MgATP>MgGTP>MgITP>MgXTP approximately MgUTP approximately MgCTP. The affinity of MgATP for HPPK (K(d)=2.6+/-0.06 microM) is 260-fold higher than that of MgGTP and more than 1000-fold higher than those of the other nucleoside triphosphates, indicating that HPPK is highly specific with respect to the base moiety of the nucleotide. The affinity of ATP for HPPK in the presence of Mg(2+) is 15 times that in the absence of Mg(2+), indicating that the metal ion is important for the binding of the nucleotide. Removal of the gamma phosphate from MgATP reduces its affinity for HPPK by a factor of approximately 21. The affinity of AMP for HPPK is about one third that of ADP and almost the same as that of adenosine. The result suggests that among the three phosphoryl groups of MgATP, the gamma-phosphoryl group is most critical for binding to HPPK and the alpha-phosphoryl group contributes little to the binding of the nucleotide. The affinity of MgATP is 18 times that of MgdATP, indicating that the 2'-hydroxyl group of MgATP is also important for binding. van't Hoff analysis suggests that binding of MgATP is mainly driven by enthalpy at 25 degrees C and the entropy of binding is also in favor of the formation of the HPPK.MgATP complex. PMID- 10825541 TI - The ecto-5'-nucleotidase subunits in dimers are not linked by disulfide bridges but by non-covalent bonds. AB - It has long been considered that ecto-5'-nucleotidase (eNT) dimers consist of subunits linked by disulfide bonds. Hydrophilic (6.7S) and amphiphilic (4.0S) dimers were separated by sedimentation analysis of eNT purified from bull seminal plasma. Hydrophilic (4. 2S) and amphiphilic (2.6S) eNT monomers were obtained after reduction of disulfide bonds in dimers. The amphiphilic eNT dimers or monomers were converted into their hydrophilic variants with phosphatidylinositol specific phospholipase C. SDS-PAGE plus Western blot showed 68 kDa subunits, regardless of the addition of beta-mercaptoethanol to the SDS mixture. Active eNT monomers were obtained by addition of 1 M guanidinium chloride (Gdn) to dimers, and unfolded subunits by addition of 4 M Gdn. The results unambiguously demonstrate that the subunits in eNT dimers are not linked by disulfide bridges, but by non-covalent bonds, and that dissociation precedes inactivation and unfolding. PMID- 10825542 TI - Calorimetric evidence for a native-like conformation of hen egg-white lysozyme dissolved in glycerol. AB - Hen egg-white lysozyme, lyophilized from aqueous solutions of different pH (from pH 2.5 to 10.0) and then dissolved in water and in anhydrous glycerol, has been studied by high-sensitivity differential scanning microcalorimetry over the temperature range from 10 to 150 degrees C. All lysozyme samples exhibit a cooperative conformational transition in both solvents occurring between 10 and 100 degrees C. The transition temperatures in glycerol are similar to those in water at the corresponding pHs. The transition enthalpies in glycerol are substantially lower than in water but follow similar pH dependences. The transition heat capacity increment in glycerol does not depend on the pH and is 1.25+/-0.31 kJ mol(-1) K(-1), which is less than one fifth of that in water (6. 72+/-0.23 kJ mol(-1) K(-1)). The thermal transition in glycerol is reversible and equilibrium, as demonstrated for the pH 8.0 sample, and follows the classical two state mechanism. In contrast to lysozyme in water, the protein dissolved in glycerol undergoes an additional, irreversible cooperative transition with a marginal endothermic heat effect at temperatures of 120-130 degrees C. The transition temperature of this second transition increases with the heating rate which is characteristic of kinetically controlled processes. Thermodynamic analysis of the calorimetric data reveals that the stability of the folded conformation of lysozyme in glycerol is similar to that in water at 20-80 degrees C but exceeds it at lower and higher temperatures. It is hypothesized that the thermal unfolding in glycerol follows the scheme: N ifho-MG-->U, where N is a native-like conformation, ho-MG is a highly ordered molten globule state, and U is the unfolded state of the protein. PMID- 10825543 TI - Inhibition of serine proteinases from human blood clotting system by squash inhibitor mutants. AB - A series of six CMTI I variants mutated in the P(2)-P(4)' region of the canonical binding loop were used to probe the role of single amino acid substitutions on binding to the following human proteinases involved in blood clotting: plasmin, plasma kallikrein, factors X(a) and XII(a). The mutants were expressed as fusion proteins with the LE1413 hydrophobic polypeptide in Escherichia coli, purified from inclusion bodies, followed by cyanobromide cleavage and refolding. The mutants inhibited the proteinases with the association constants in the range 10(3)-10(9) M(-1). Inhibition of plasma kallikrein and factors X(a) and XII(a) could be improved up to 30-fold by single mutations. In contrast, neither of the introduced mutations increased inhibitory properties of CMTI I against plasmin. Additionally, using two inhibitors of natural origin, CMTI I (P(1) Arg) and CPTI II (P(1) Lys), we determined the effect of Lys-->Arg on binding to four proteinases. With the exception of plasmin (no effect), P(1) Arg resulted in up to 30-fold stronger binding than P(1) Lys. PMID- 10825544 TI - Domain-domain interface packing at conserved Trp-20 in class alpha glutathione transferase impacts on protein stability. AB - The folding and assembly of the dimeric glutathione transferases (GST) involves the association of two structurally distinct domains per subunit. A prominent and conserved domain-domain interaction in class alpha GSTs is formed by the packing of the indole side chain of Trp-20 from domain I into a hydrophobic pocket in domain II. Stability studies have shown that partial dissociation of the domains near Trp-20 occurs as an initial fast event during the unfolding kinetics of human GSTA1-1 (Wallace et al., Biochemistry 37 (1998) 5320-5328; Wallace et al., Biochem. J. 336 (1998) 413-418). The contribution of Trp-20 toward stabilising the domain-domain interface was investigated by mutating it to either a phenylalanine (W20F) or alanine (W20A) and determining the functionality (catalysis and non-substrate ligand binding) and stability (thermal- and urea induced denaturation) of the mutant proteins. The replacement of Trp-20 did not impact on the protein's gross structural properties. Functionally, the W20F was non-disruptive, whereas the cavity-creating W20A mutation was. Both mutants destabilised the native state with W20A exerting the greatest effect. Reduced m values as well as the protein concentration dependence of the urea unfolding transitions for W20F GSTA1-1 suggest the presence of a dimeric intermediate at equilibrium that is not observed with wild-type protein. Unfolding kinetics monitored by stopped-flow tyrosine fluorescence was mono-exponential and corresponded to the global unfolding of the protein during which the dimeric intermediate unfolds to two unfolded monomers. The similar unfolding kinetics data for wild-type and W20F A1-1 indicates that the global unfolding event was not affected by amino acid replacement. We propose that the packing interactions at the conserved Trp-20 plays an important role in stabilising the intrasubunit domain I-domain II interface of class alpha GSTs. PMID- 10825545 TI - Molecular characterization of a dimeric intracellular maltogenic amylase of Bacillus subtilis SUH4-2. AB - An additional amylase besides the typical alpha-amylase was detected in the cytoplasm of Bacillus subtilis SUH4-2, an isolate from Korean soil. The corresponding gene encoded a maltogenic amylase, which hydrolyzed cyclodextrin or starch to maltose and glucose; pullulan to panose; acarbose to glucose and acarviosine-glucose. Maltogenic amylase of B. subtilis SUH4-2 transferred sugar molecules to form various branched oligosaccharides upon the hydrolysis of substrates. The enzyme existed in a monomer-dimer equilibrium with a molar ratio of 3:2 in 50 mM KH(2)PO(4)-NaOH buffer (pH 7.0). The maltogenic amylase is most likely to be associated with carbohydrate metabolism in the cytoplasm, since the nucleotide sequence of the gene was highly homologous to the yvdF gene of B. subtilis 168, which is located in a gene cluster involved in maltose/maltodextrin utilization. PMID- 10825546 TI - Dimerization of signalling modules of the EvgAS and BvgAS phosphorelay systems. AB - Biophysical and biochemical properties of signalling proteins or domains derived from the unorthodox EvgAS and BvgAS two-component phosphorelay systems of Escherichia coli and Bordetella pertussis were investigated. Oligomerization of the effector proteins EvgA and BvgA and of truncated EvgS and BvgS derived signalling proteins containing the receiver and histidine containing phosphotransfer (HPt) domains or comprising only the HPt domains were characterized by native gel electrophoresis, gel permeation experiments and analytical ultracentrifugation. The results obtained by the different methods are consistent with non-phosphorylated EvgA and BvgA proteins being dimers in solution with a dissociation constant significantly below 1 microM. In contrast, all sensor derived domains of EvgS and BvgS were observed to be monomers in vitro. No indications for a phosphorylation induced stimulation of oligomerization of the C-terminal histidine kinase domains could be detected. In agreement with these data, surface plasmon resonance studies revealed a 2:1 stoichiometry in the interaction of EvgA with the immobilized EvgS HPt domain and an affinity constant of 1. 24x10(6) M(-1). PMID- 10825547 TI - Recent advances in mucosal vaccine development. AB - Proper stimulation of the mucosal immune system is critical for the effective protection of mucosal surfaces against colonization and invasion of infectious agents. This requires administration of vaccine antigens directly to various mucosal sites. Due to the low absorption efficiency of mucosally delivered vaccines, however, almost all of the currently marketed vaccines are administered parentally. In addition, sub-optimal immune responses are frequently induced by mucosal immunization and the use of mucosal adjuvants is commonly required. As a result, development of successful mucosal vaccines depends largely on the improvement of mucosal antigen delivery and on the discovery of new and effective mucosal adjuvants. In this review, recent advances in both areas are briefly discussed. PMID- 10825548 TI - A biodegradable multiblock co-polymer derived from an alpha, omega bis(methylamino)peptide and an alpha, omega-bis(oxiranylmethyl)poly(ethylene glycol). AB - A novel peptide containing the lysosomally degradable sequence GlyPheLeuGly and a sequence-inverting unit has been prepared. This peptide presents the methylamino groups of sarcosine (N-methylglycine) at both termini for co-polymerisation with alpha, omega-bis(oxiranylmethoxy)poly(ethylene glycol) of mean M(w) 1650. Gel permeation chromatographic analysis showed the presence of a mixture of oligomers. Preliminary degradation studies showed that these oligomers are cleaved by cathepsin B, an important lysosomal enzyme. PMID- 10825549 TI - Endothelial cell seeding of (heparinized) collagen matrices: effects of bFGF pre loading on proliferation (after low density seeding) and pro-coagulant factors. AB - Endothelial cell seeding to improve the performance of small-diameter vascular grafts requires a suitable substrate, such as crosslinked collagen. In addition to providing a suitable substrate for adhesion and growth of endothelial cells, proliferation of seeded endothelial cells can be enhanced by local, sustained release of basic fibroblast growth factor (bFGF, a heparin-binding growth factor for endothelial cells). We have previously shown that collagen crosslinked using N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide (EDC) and N-hydroxysuccinimide (NHS) supports adhesion and proliferation of human umbilical vein endothelial cells (HUVECs). In the present study, HUVECs were seeded on (heparinized) EDC/NHS crosslinked collagen, pre-loaded with bFGF. Proliferation of HUVECs on (heparinized) crosslinked collagen increased with increasing amounts of pre loaded bFGF. The minimal cell-seeding density required for proliferation proved to be very low after pre-loading the substrates with bFGF, and was 4-fold lower for heparinized crosslinked collagen compared to crosslinked collagen (250 versus 1000 cells/cm(2)). Pro-coagulant properties (von Willebrand factor secretion and tissue factor expression) of HUVECs seeded on (heparinized) crosslinked collagen, with or without pre-loading of bFGF, were comparable to those of HUVECs on TCPS. It is concluded that heparinized, EDC/NHS-crosslinked collagen pre-loaded with bFGF is a candidate matrix for in vivo endothelial cell seeding of synthetic vascular graft materials. PMID- 10825550 TI - Sulfobutylated poly(vinyl alcohol)-graft-poly(lactide-co-glycolide)s facilitate the preparation of small negatively charged biodegradable nanospheres. AB - The manufacturing conditions for small nanoparticles (NP) in the range of 100-500 nm are difficult to control. Novel biodegradable, brush-like branched polyesters with a negatively charged hydrophilic backbone, poly(2-sulfobutyl-vinyl alcohol) g-poly(lactide-co-glycolide), facilitate their preparation by a modified solvent displacement procedure. Furthermore, the structure and the surface properties of the colloidal systems are investigated. NP were characterized by photon correlation spectroscopy (PCS), zeta-potential measurement (ZPM), particle charge detection (PCD), nuclear magnetic resonance spectroscopy (NMR) and transmission electron microscopy (TEM). Varying the manufacturing conditions NP with mean diameters of about 100 up to 500 nm and, depending on polymer composition, negatively charged surfaces were obtained. The NP visualized by TEM showed smooth surfaces. Furthermore, surface characterization and NMR studies suggested a core/corona structure of the particles. This study demonstrates that a simple solvent displacement technique can be used for the reproducible preparation of discrete NP with defined negatively charged surfaces and narrow size distributions. These NP may have potential for peroral or parenteral protein delivery systems. PMID- 10825551 TI - Incorporation of protein in PLG-microspheres with retention of bioactivity. AB - The enzyme urease was incorporated into poly(lactide-co-glycolide) microspheres using a double emulsion solvent removal technique. Ethyl acetate was used as organic solvent since it is less toxic than the more commonly used methylene chloride. The effect of the two solvents on urease was compared. Although this preparation technique is well established, it is often associated with reduced bioactivity and low entrapment efficiency of proteins. In order to retain a high degree of bioactivity, the well known protein stabilisers: sucrose, trehalose and poloxamer 407, were added to the urease in the preparation. The bioactivity of the entrapped urease was reduced more by methylene chloride than by ethyl acetate. The gelled form of poloxamer was shown to highly favour the retention of bioactivity, demonstrated by an increase of 41% compared to preparations without poloxamer. Moreover, the presence of poloxamer strongly increased the in vitro release rate of urease from the microspheres. The entrapment efficiency was increased by 44% using the sugars in the preparation. These results clearly show the great potential of small quantities of additive in the formulation to control the properties of the microspheres. The amount and type of additive could be adjusted according to the therapeutic application of the preparation. PMID- 10825552 TI - Controlled transdermal iontophoresis by ion-exchange fiber. AB - The objective of this study was to assess the transdermal delivery of drugs using iontophoresis with cation- and anion-exchange fibers as controlled drug delivery vehicles. Complexation of charged model drugs with the ion-exchange fibers was studied as a method to achieve controlled transdermal drug delivery. Drug release from the cation-exchange fiber into a physiological saline was dependent on the lipophilicity of the drug. The release rates of lipophilic tacrine and propranolol were significantly slower than that of hydrophilic nadolol. Permeation of tacrine across the skin was directly related to the iontophoretic current density and drug concentration used. Anion-exchange fiber was tested with anionic sodium salicylate. The iontophoretic flux enhancement of sodium salicylate from the fiber was substantial. As the drug has to be released from the ion-exchange fiber before permeating across the skin, a clear reduction in the drug fluxes from the cationic and anionic fibers were observed compared to the respective fluxes of the drugs in solution. Overall, the ion-exchange fibers act as a drug reservoir, controlling the release and iontophoretic transdermal delivery of the drug. PMID- 10825553 TI - Experimental investigation and mathematical modeling of Pluronic F127 gel dissolution: drug release in stirred systems. AB - We have examined the dissolution of Pluronic F127 gels in a USP dissolution apparatus under stirred conditions, and simultaneously monitored the release of model drugs from these gels. The drugs selected were propranolol HCl, metronidazole and cephalexin. Our results show that drug release is zero-order and is controlled by the dissolution of the gel for all the drugs, under various conditions of temperature, F127 concentration, drug concentration, and for stirring speeds between 20 and 80 rpm. The addition of inorganic salts has no significant effect on dissolution rate or drug release. Increasing F127 concentration in the gel decreases gel dissolution and drug release rates. We have developed a predictive mathematical model based on the assumption that uptake of water into the gel and subsequent disentanglement of F127 micelles control gel dissolution. There is good agreement between experimental results and model predictions for stirring speeds above 20 rpm. As stirring speed is decreased to 20 rpm and below, there are discrepancies between actual and predicted values, presumably due to a significant diffusion component that contributes to drug release. PMID- 10825554 TI - Polymer-induced leakage of cations from dioleoyl phosphatidylcholine and phosphatidylglycerol liposomes. AB - The amphipathic polyacid, poly(2-ethylacrylic acid) (PEAA) has recently been shown to form fluctuating channels in patch-clamp measurements of phospholipid bilayers ?J.C. Chung, D.J. Gross, J.L. Thomas, D.A. Tirrell, L.R. Opsahl-Ong, Macromolecules 29 (1996) 4636-4641.. To explore this phenomenon further, we have quantified the PEAA-mediated pH-dependent release of sodium and calcium ions from phospholipid vesicles. Permeability to calcium increases linearly with polymer concentration and exponentially with decreasing pH. Permeabilization of negatively charged phosphatidylglycerol (PG) liposomes occurs to a similar extent and with a similar pH dependence to that of zwitterionic phosphatidylcholine (PC) liposomes, implying that a charge neutral species is responsible for the leakage. The pH dependence of leakage shows that the cooperative protonation of from three to five carboxylate anions is required for permeabilization. Such neutralization could result in a neutral segment of polymer chain of sufficient length to span the bilayer. PMID- 10825555 TI - Reduction in skin permeation of N,N-diethyl-m-toluamide (DEET) by altering the skin/vehicle partition coefficient. AB - Reported adverse side effects after using N,N-diethyl-m-toluamide (DEET) containing mosquito repellent products appear to be the result of significant absorption of DEET through human skin. The overall objective was to develop formulations of DEET with significantly reduced permeation using the basic principles and model of skin permeation based on Fick's laws of diffusion at steady state. Ternary phase diagrams of DEET with water and semipolar solvents, ethanol, PG and PEG 400, showed an increase in the aqueous solubility of DEET. This resulted in a linear decline in octanol/water PC with an increase in the concentration of the solvent. Permeation of DEET across human skin was studied from vehicles containing various amounts of PG and PEG 400 using an infinite dose technique and Franz diffusion cell. DEET's flux reduced with increasing PG concentration and the flux from 90% PG was 9.9+/-2.1 microg/cm(2) h, 6-fold lower than flux of pure DEET control, 63.2+/-24.5 microg/cm(2) h. Flux was reduced 6 fold from 60% PEG 400 solution, and permeation of DEET was totally prevented from 90% PEG 400 which was very viscous. However, a combination of 60% PEG 400 with 30% PG not only reduced permeation 9-fold but was suitable as a vehicle for formulation. The decrease in flux and permeability of DEET with increasing concentration of solvent appeared to be a direct result of decrease in skin/vehicle PC and octanol/water PC. This study clearly demonstrates that alternative formulations can be developed for DEET aimed at reduced permeation and toxicity unlike the current formulations some of which contain ethanol which has been shown to enhance permeation of DEET. A similar approach can be used for developing formulations of other industrial and occupational agents to prevent their skin permeation when a user may be exposed to them. PMID- 10825556 TI - Evaluation of poly(acrylic acid-co-ethylhexyl acrylate) films for mucoadhesive transbuccal drug delivery: factors affecting the force of mucoadhesion. AB - Based on the premise that similar surface properties between the adhesive and the substrate would yield a strong adhesive bond, copolymers of acrylic acid (AA) and 2-ethylhexyl acrylate (EHA), P(AA-co-EHA), were designed and synthesized for buccal mucoadhesion. A series of linear copolymers with varying feed ratios of the two monomers (AA and EHA) were synthesized through free radical copolymerization at 69+/-0.5 degrees C using azobis(isobutyronitrile) (AIBN) as initiator. The reactions were carried out in THF under nitrogen for 24 h. The glass transition temperatures, T(g), of the copolymers were determined using DSC. The adhesion studies were conducted to determine the effects of copolymer composition, contact time between the substrate and the adhesive, and crosshead speed on mucoadhesive performance of the copolymer films using a computer interfaced Instron material testing system. The glass transition temperature of the copolymers decreased with increasing EHA content. Wet glass surface as substrate was shown not to be a good substrate model for adhesion determination studies. The copolymer composed of 46:54 mol.% AA:EHA (an almost 1:1 ratio in the repeat units) yielded the highest mucoadhesive force in contact with porcine buccal mucosa which was significantly greater (P<0.05) than that of poly(acrylic acid) (PAA) (used as positive control). The mucoadhesive force for all copolymers studied was significantly (P<0.05) greater than that of the negative control (backing material without copolymer film) except for the EHA homopolymer. Crosshead speed increased mucoadhesive force linearly and had a more pronounced effect on the mucoadhesive performance than time of contact between the adhesive and the substrate. PMID- 10825557 TI - Microspheres for protein delivery prepared from amphiphilic multiblock copolymers. 1. Influence of preparation techniques on particle characteristics and protein delivery. AB - The entrapment of lysozyme in amphiphilic multiblock copolymer microspheres by emulsification and subsequent solvent removal processes was studied. The copolymers are composed of hydrophilic poly(ethylene glycol) (PEG) blocks and hydrophobic poly(butylene terephthalate) (PBT) blocks. Direct solvent extraction from a water-in-oil (w/o) emulsion in ethanol or methanol did not result in the formation of microspheres, due to massive polymer precipitation caused by rapid solvent extraction in these non-solvents. In a second process, microspheres were first prepared by a water-in-oil-in-water (w/o/w) emulsion system with 4% poly(vinyl alcohol) (PVA) as stabilizer in the external phase, followed by extraction of the remaining solvent. As non-solvents ethanol, methanol and mixtures of methanol and water were employed. However, the use of alcohols in the extraction medium resulted in microspheres which gave an incomplete lysozyme release at a non-constant rate. Complete lysozyme release was obtained from microspheres prepared by an emulsification-solvent evaporation method in PBS containing poly(vinyl pyrrolidone) (PVP) or PVA as stabilizer. PVA was most effective in stabilizing the w/o/w emulsion. Perfectly spherical microspheres were produced, with high protein entrapment efficiencies. These microspheres released lysozyme at an almost constant rate for approximately 28 days. The reproducibility of the w/o/w emulsion process was demonstrated by comparing particle characteristics and release profiles of three batches, prepared under similar conditions. PMID- 10825558 TI - Microspheres for protein delivery prepared from amphiphilic multiblock copolymers. 2. Modulation of release rate. AB - Amphiphilic multiblock copolymers, based on hydrophilic poly(ethylene glycol) (PEG) blocks and hydrophobic poly(butylene terephthalate) (PBT) blocks were used as matrix material for protein-loaded microspheres. The efficiency of lysozyme entrapment by a double emulsion method was found to depend on the swelling behavior of the polymers in water and decreased from 100% for polymers with a degree of swelling of less than 1.8 to 11% for PEG-PBT copolymers with a degree of swelling of 3.6. The particle size could be controlled by varying the concentration of the polymer solution used in the microsphere preparation. An increase in the polymer concentration resulted in a proportional increase in the particle size. The in vitro release profiles of the encapsulated model protein lysozyme could be precisely tailored by variation of the copolymer composition and the size of the microspheres. Both a slow continuous release of lysozyme, and a fast release which was completed within a few days could be obtained. The release behavior, attributed to a combination of diffusion and polymer degradation, could be described by a previously developed model. PMID- 10825559 TI - Correlation between drug release kinetics from proteineous matrices and protein folding: elasticity and compressibility study. AB - Naproxen sodium (NS) release mechanism from proteineous matrices based on egg albumin (EA) and bovine serum albumin (BSA) was investigated by several physico chemical methods. The gel strength, modulus of elasticity and erosion properties of the matrices were studied and correlated with drug release kinetics. The results revealed that NS release rate from EA and BSA matrices was markedly different, indicating the significant role of protein nature and conformation on matrix behavior. Unexpectedly it was found that incorporation of NS to EA matrix increased gel strength and modulus of elasticity and decreased matrix erosion. This effect was dependent on NS concentration in the matrix. In contrast to EA, BSA behaved as a non-gelling matrix and was unable to retard drug release because of its high solubility. The influence of NS on protein folding and compressibility in protein solutions was studied using densitometric and ultrasonic techniques. Adiabatic compressibility measurements revealed that NS caused unfolding of EA, an effect which led to a decrease in EA intrinsic compressibility and the exposure of atomic side groups buried in protein interior. Unfolding of EA led to an increase of modulus of elasticity in solution (measured by ultrasonic velocimetry technique) which is in correlation with the modulus of elasticity measurements of gelled tablets (measured by Instron). In concentrated EA solutions, the results showed a large increase in EA compressibility and ultrasonic absorption in the presence of NS indicating a strong aggregation of the denatured state of EA. Regarding BSA, the results suggested that NS affected the packing of the protein interior, transforming it to a molten globule intermediate state, an effect that led to an increase in BSA compressibility. At high BSA concentrations, aggregation of the molten globule state was observed as indicated by an increase of BSA compressibility and ultrasonic absorption values. PMID- 10825560 TI - Oral sustained delivery of theophylline using in-situ gelation of sodium alginate. AB - Gels formed in situ following oral administration of aqueous solutions of sodium alginate (1.0-2.0%w/v) to rats were evaluated as sustained release vehicles for the delivery of theophylline. The liquid formulation contained calcium ions in complexed form, the release of which in the acidic environment of the stomach caused gelation of the alginate. Bioavailability of theophylline from alginate gels formed by in situ gelation in the rat stomach was increased by 1.3-2-fold in rats for alginate concentrations of 2.0 to 1.0%w/v respectively compared with that from a proprietary oral sustained release formulation containing an identical drug concentration. There was no significant difference in the mean residence times of theophylline when administered by these two vehicles. PMID- 10825561 TI - Sustained release of recombinant human insulin-like growth factor-I for treatment of diabetes. AB - Recombinant human insulin-like growth factor-I (rhIGF-I) was found to improve glycemic control and enhance insulin sensitivity in patients with a syndrome of severe insulin resistance. Therefore, the protein may be considered as an alternative therapy in the treatment of diabetes when the patients become insensitive to insulin treatment. Because the protein was administered twice per day in the clinical trials, a sustained release polylactic-co-glycolic acid (PLGA) formulation for rhIGF-I with low initial burst (<20%), maximum possible protein loading (15-20%) and a continuous release of 1-2 weeks may provide greater patient convenience and compliance. The protein was encapsulated in PLGA for sustained release using a spray freeze-drying technique. Formulation parameters such as protein loading, polymer end group, and the presence of zinc carbonate were studied for their effects on in vitro release of rhIGF-I from PLGA microspheres. As the protein loading was increased, the initial burst increased. Due to the hydrophilic properties of the polymers, rhIGF-I encapsulated in unblocked PLGA (free acid end groups) gave a lower initial burst and a more steady-state release profile than the blocked PLGA (hydrocarbon end groups) with the same protein loading and PLGA molecular weight. At 15% w/w protein loading, the addition of 6% w/w zinc carbonate as a protein release modifier to the unblocked PLGA (12 kDa) decreased the initial burst of rhIGF-I. Therefore, a formulation consisting of 15% rhIGF-I and 6% zinc carbonate in 12 kDa, unblocked 50:50 PLGA can provide the required release characteristics in vitro. Rat studies revealed that rhIGF-I in this formulation was released in vivo at a rate which was comparable to that observed in vitro. These studies demonstrate the potential for a sustained release, 14-day formulation for rhIGF-I. PMID- 10825562 TI - In vitro studies and modeling of a controlled-release device for root canal therapy. AB - Endodontic disease is caused primarily by bacteria that interact with periradicular host tissues. Therefore, treatment of endodontic disease aims at the exclusion of bacteria from the root canal system. This work focused on in vitro studies and modeling of a controlled-release device for delivering antimicrobial agents in root canals. A cylindrical, needle-shaped device was prepared consisting of a matrix core and a polymer coating, loaded with 30-45% chlorhexidine (CHX). The composition of the core, a blend of water-permeable polymers, and the thickness of the coating were tailored to impart various release rates. A relatively steady release rate for over 40 days after an initial burst was achieved using a formulation for long-term release, which is desirable for establishing and maintaining the necessary therapeutic levels. Mathematical models were developed for both in vitro and in vivo drug release into a liquid of limited volume, taking into account a moving boundary of the dispersed drug and a time-dependent boundary condition. A concentration-dependent effective diffusion coefficient was used to count increased porosity as the solid drug had dissolved. The finite element method and computer programs were applied to solve the differential equations and predict the in vitro and in vivo release kinetics. The model prediction agreed well with the in vitro experimental data and provided guidance for designing the device for in vivo release in root canals. The result of in vitro antimicrobial tests, performed using a bovine tooth model, suggested that the device was effective in reducing growth of microbes. PMID- 10825563 TI - Monolithic osmotic tablet system for nifedipine delivery. AB - The monolithic osmotic tablet system, which is composed of a monolithic tablet coated with cellulose acetate (CA) membrane drilled with two orifices on both side surfaces, has been described. The influences of tablet formulation variables including molecular weight (MW) and amount of polyethylene oxide (PEO), amount of potassium chloride (KCl), and amount of rice starch as well as nifedipine loading have been investigated. The optimal tablet formulation and the osmotic-suspending co-controlled delivery mechanisms have been proposed. Orifice size and membrane variables including nature and amount of plasticizers as well as thickness on drug release have also been studied. The in vitro release profiles of the optimal system have been evaluated in various release media and different agitation rates, and compared with commercialized conventional capsule and push-pull osmotic tablet. It was found that PEO with MW of 300000 g/mol was suitable to be thickening agent, both amount of KCl and amount of PEO had comparable and profoundly positive effects, while nifedipine loading had a strikingly negative influence on drug release. It could be found that the optimal orifice size was in the range of 0.25-1.41 mm. It has also been observed that hydrophilic plasticizer polyethylene glycol (PEG) improved drug release, whereas hydrophobic plasticizer triacetin depressed drug release when they were incorporated in CA membrane. The monolithic osmotic tablet system was found to be able to deliver nifedipine at the rate of approximate zero-order up to 24 h, independent of both environmental media and agitation rate, and substantially comparable with the push-pull osmotic tablet. The monolithic osmotic tablet system was simple to be prepared as exempting from push layer and simplifying in the orifice drilling compared with the push-pull osmotic tablet. The monolithic osmotic tablet system may be used in drug controlled delivery field, especially suitable for water-insoluble drugs. PMID- 10825564 TI - In vivo and in vitro release of lysozyme from cross-linked gelatin hydrogels: a model system for the delivery of antibacterial proteins from prosthetic heart valves. AB - Prosthetic valve endocarditis may be reduced by the local delivery of antibacterial proteins from the Dacron sewing ring of a prosthetic heart valve. Dacron discs were treated with a carbon dioxide gas plasma to improve the hydrophilicity and thereby enabling homogeneous impregnation with gelatin type B. The gelatin samples were cross-linked to different degrees using various amounts of water-soluble carbodiimide (EDC) and N-hydroxysuccinimide (NHS). Lysozyme, a model protein for antibacterial proteins, was loaded into (non)-cross-linked gelatin gels incorporated in Dacron, or adsorbed onto non-treated and gas plasma treated Dacron. The in vivo lysozyme release was measured after subcutaneous implantation of lysozyme-loaded samples in rats. The lysozyme content of the samples, and the lysozyme level of the surrounding tissue were determined at different explantation times (ranging from 6 h up to 1 week). For cross-linked gelatin gels, the lysozyme tissue level was elevated up to 2 days after implantation. In vitro release was measured using agarose medium or phosphate buffer. Lysozyme release in buffer solution under sink conditions was in good agreement with the in vivo lysozyme release profiles, and therefore considered a good model to describe in vivo release characteristics. The release was modelled with a solution of Fick's second law of diffusion using the appropriate boundary conditions. In this way the lysozyme concentration in the gel and the surrounding tissue as a function of time and distance was obtained. The presence of cross linked gelatin in Dacron did lead to an increased uptake of lysozyme and a delayed release during 30 h after implantation, whereas a burst release took place from Dacron, gas plasma-treated Dacron, or Dacron containing non-cross linked gelatin. PMID- 10825565 TI - Electroporation-mediated delivery of 3'-protected phosphodiester oligodeoxynucleotides to the skin. AB - The feasibility of topical delivery in the skin of 3' end modified phosphodiester oligonucleotides using electroporation was investigated. Experiments were performed in vitro, using hairless rat skin. Five pulses of (200 V, 450 ms) were applied. The 3' end modifications of the 15 mer oligonucleotide were: (1) 3' aminohexyl, (2) biotin, with a triethyleneglycol arm, (3) methylphosphonate links between nucleotides 13, 14 and 15, and (4) 2-O-methyl nucleotides at 13, 14 and 15 positions. All the modifications were efficient to protect the oligonucleotides against degradation in the skin. Electroporation increased the topical delivery of the 3' end-modified phosphodiesters by two orders of magnitude compared to passive diffusion, without significant differences between the derivatives. Oligonucleotide concentrations in the range of 1 microm could be achieved in the viable skin. The delivery of a phosphorothioate congener was lower than phosphodiester delivery due to the interaction of phosphorothioate with the stratum corneum. Consequently, 3' end-protected phosphodiesters could be an interesting alternative to phosphorothioate oligonucleotides for topical treatment of cutaneous diseases. PMID- 10825566 TI - Novel anionic microparticles are a potent adjuvant for the induction of cytotoxic T lymphocytes against recombinant p55 gag from HIV-1. AB - Microparticles with entrapped antigens have recently been shown to possess significant potential as vaccine delivery systems and adjuvants. However, the potential of microparticles as adjuvants has been seriously limited by the common problem of degradation and denaturation of antigens following encapsulation and release. To overcome these problems, we have developed a novel way to use microparticles as adjuvants, by the adsorption of proteins onto their surface. Anionic microparticles were prepared through the inclusion of an anionic detergent, sodium dodecyl sulphate (SDS), in the microparticle preparation process. The anionic microparticles were capable of the efficient and reproducible adsorption of recombinant p55 gag protein from HIV-1. Microparticles with adsorbed p55 were capable of inducing potent cytotoxic T lymphocyte responses in mice following intramuscular immunization. In addition, the microparticles also exhibited a potent adjuvant effect for antibody induction against p55. PMID- 10825567 TI - Design, characterisation and preliminary clinical evaluation of a novel mucoadhesive topical formulation containing tetracycline for the treatment of periodontal disease. AB - This study describes the formulation, characterisation and preliminary clinical evaluation of mucoadhesive, semi-solid formulations containing hydroxyethylcellulose (HEC, 1-5%, w/w), polyvinylpyrrolidine (PVP, 2 or 3%, w/w), polycarbophil (PC, 1 or 3%, w/w) and tetracycline (5%, w/w, as the hydrochloride). Each formulation was characterised in terms of drug release, hardness, compressibility, adhesiveness (using a texture analyser in texture profile analysis mode), syringeability (using a texture analyser in compression mode) and adhesion to a mucin disc (measured as a detachment force using the texture analyser in tensile mode). The release exponent for the formulations ranged from 0.78+/-0.02 to 1. 27+/-0.07, indicating that drug release was non diffusion controlled. Increasing the concentrations of each polymeric component significantly increased the time required for 10 and 30% release of the original mass of tetracycline, due to both increased viscosity and, additionally, the unique swelling properties of the formulations. Increasing concentrations of each polymeric component also increased the hardness, compressibility, adhesiveness, syringeability and mucoadhesion of the formulations. The effects on product hardness, compressibility and syringeability may be due to increased product viscosity and, hence, increased resistance to compression. Similarly, the effects of these polymers on adhesiveness/mucoadhesion highlight their mucoadhesive nature and, importantly, the effects of polymer state (particularly PC) on these properties. Thus, in formulations where the neutralisation of PC was maximally suppressed, adhesiveness and mucoadhesion were also maximal. Interestingly, statistical interactions were primarily observed between the effects of HEC and PC on drug release, mechanical and mucoadhesive properties. These were explained by the effects of HEC on the physical state of PC, namely swollen or unswollen. In the preliminary clinical evaluation, a formulation was selected that offered an appropriate balance of the above physical properties and contained 3% HEC, 3% PVP and 1% PC, in addition to tetracycline 5% (as the hydrochloride). The clinical efficacy of this (test) formulation was compared to an identical tetracycline-devoid (control) formulation in nine periodontal pockets (>/=5 mm depth). One week following administration of the test formulation, there was a significant improvement in periodontal health as identified by reduced numbers of sub-gingival microbial pathogens. Therefore, it can be concluded that, when used in combination with mechanical plaque removal, the tetracycline-containing semi solid systems described in this study would augment such therapy by enhancing the removal of pathogens, thus improving periodontal health. PMID- 10825568 TI - Solid dosage form preparations from oily medicines and their drug release. Effect Of degree of surface-modification of silica gel on the drug release from phytonadione-loaded silica gels. AB - A surface-modified silica gel was produced to improve the surface affinity to an oily medicine, phytonadione (VK1). The effect of the degree of surface modification of the silica gel on the drug release behavior from the silica porous matrix was investigated. The silica gels were surface-modified using the silan coupling agent, 3-methacryloxypropyltrimethoxysilane (C7), octadecyltriethoxysilane (C18), or 3,3,3-trifluoropropyltrimethoxysilane (F3). A mixture of VK1 solution and surface-modified silica gel was evaporated under reduced pressure at room temperature, then the resulting powder was dried in vacuo. The degree of surface modification was evaluated based upon elementary analysis. The dissolution profiles of the samples were investigated in Japanese Pharmacopoeia XII, 1st fluid buffer (pH 1.2, 37+/-0.5 degrees C) containing 1.5% sodium lauryl sulfate. The FT-IR spectra of VK1-loaded surface-modified silica gels suggested that the amount of hydrogen-bonded VK1 with the silanol group on the gel surface decreased with increasing hydrophobicity of the silica gel. Since the modified group was rotating on the silica gel surface, and inhibited the adsorption of VK1 to the surface, the attractive molecular interaction between VK1 and the silica gel surface might decrease with increasing length of the modified functional group. However, the characteristics of the affinity of VK1 to the functional groups significantly differed among the groups. The VK1 release from the modified silica gels was initially rapid, slowed markedly after 1 h, and continued for more than 24 h. The amount of VK1 released from the modified surface silica gels by C7, C18 or F3 increased with increasing density of the surface modification group. The mean drug release moment (MDT) decreased with an increase in surface-modified group density. PMID- 10825569 TI - Controlled release of platelet-derived growth factor-BB from chondroitin sulfate chitosan sponge for guided bone regeneration. AB - Platelet-derived growth factor-BB (PDGF-BB) releasing porous chondroitin-4 sulfate (CS)-chitosan sponge was designed with an aim of controlling growth factor delivery in order to improve bone formation. Porous CS-chitosan sponge was fabricated by freeze drying and crosslinking aqueous CS-chitosan solution. PDGF BB was incorporated into the CS-chitosan sponge by soaking CS-chitosan sponge into the PDGF-BB solution. CS-chitosan sponge retained a porous structure with a 150-200-microm pore diameter that was suitable for cellular migration and osteoid ingrowth. Release rate of PDGF-BB could be controlled by varying the composition of CS in the sponge or initial loading content of PDGF-BB. CS-chitosan sponge induced increased osteoblast migration and proliferation as compared with chitosan sponge alone. Furthermore, the release of PDGF-BB from CS-chitosan sponge significantly enhanced osteoblast proliferation. These results suggest that PDGF-BB-releasing CS-chitosan sponge may be beneficial to enhance bone cell adaptation and regenerative potential when applied in wound sites. PMID- 10825570 TI - Rheological, mechanical and membrane penetration properties of novel dual drug systems for percutaneous delivery. AB - In this study it has been demonstrated that mixtures of two solid drugs, ibuprofen and methyl nicotinate, with different but complementary pharmacological activities and which exist as a single liquid phase over a wide composition range at skin temperature, can be formulated as o/w emulsions without the use of an additional hydrophobic carrier. These novel dual drug systems provided significantly enhanced in vitro penetration rates through a model lipophilic barrier membrane compared to conventional individual formulations of each active. Thus, for ibuprofen, drug penetration flux enhancements of three- and 10-fold were observed when compared to an aqueous ibuprofen suspension and a commercial alcohol-based ibuprofen formulation, respectively. Methyl nicotinate penetration rates were shown to be similar for aqueous gels and emulsified systems. Mechanisms explaining these observations are proposed. Novel dual drug formulations of ibuprofen and methyl nicotinate, formulated within the liquid range at skin temperature, were investigated by oscillatory rheology and texture profile analysis, demonstrating the effects of drug and viscosity enhancer concentrations, and disperse phase type upon the rheological, mechanical and drug penetration properties of these systems. PMID- 10825571 TI - Large unilamellar vesicles as trehalose-stabilised vehicles for vaccines: storage time and in vivo studies. AB - Liposomes, as a pharmaceutical formulation must display a long shelf life. The recombinant heat-shock protein from Mycobacterium leprae (18-kDa hsp) or its N acylated derivative, when entrapped within or externally associated with large unilamellar vesicles, acts as a T-epitope source. Freeze-fracture electron microscopy shows unequivocally that trehalose avoids aggregation and fusion of these vesicles. Formulations containing trehalose retained up to 98% of the entrapped protein. The highest antibody level is obtained with formulations containing trehalose. The adjuvant effect depends on the liposomal membrane integrity. PMID- 10825573 TI - Nuclear magnetic relaxation dispersion profiles of substantia nigra pars compacta in Parkinson's disease patients are consistent with protein aggregation. AB - Nuclear Magnetic Relaxation field-cycling relaxometry is a technique, able to report on water mobility in tissues. By means of this technique, post-mortem specimens from both controls and idiopathic Parkinson's disease patients have been investigated. Results show different relaxometric behavior between the groups, which is consistent with protein aggregation in Parkinson's disease specimens. PMID- 10825574 TI - Regulation of the dual function tissue transglutaminase/Galpha(h) during murine neuromuscular development: gene and enzyme isoform expression. AB - Coagulation Factor XIII (F. VIII), a member of the transglutaminase (TGase) superfamily, is activated by thrombin, cross-links fibrin and stabilizes clots. Another member of this family, tissue TGase (tTG), having similar enzymatic activity, is implicated in neural development and synapse stabilization. Our previous studies indicated that synapse formation and maintenance at the neuromuscular junction (NMJ) involved components of the coagulation cascade in development. Others then showed that either F. XIII or tTG were localized at NMJs in a developmentally-regulated fashion. In the current studies, we addressed the temporal course of skeletal muscle tTG gene expression and found maximal expression at birth and continuing into the immediate postnatal period. Subcellular fractionation revealed a relatively constant particulate isoform of TGase activity which predominated in early embryonic muscle development. In contrast, cytosolic TGase specific activity became the major isoform in the postnatal period. The timing of muscle TGase activity correlated well with expression of tTG mRNA and we now present novel data of Tgm 2 gene expression for tTG in skeletal muscle. Confirming and extending the previous studies, TGase becomes localized at NMJs in the early, further ramifying in the late, neonatal period. These data suggest that the early pulse of particulate activity could coincide with the period of myoblast cell death in embryonic muscle. On the other hand, the peak cytosolic TGase activity occurs in the neonatal period, correlating temporally with muscle prothrombin expression during activity dependent synapse elimination and possibly the source of the enzyme localized to the NMJ extracellular matrix resulting in synaptic stabilization. PMID- 10825575 TI - Development and characterization of antibodies specific to caspase-3-produced alpha II-spectrin 120 kDa breakdown product: marker for neuronal apoptosis. AB - Alpha II-spectrin (alpha-fodrin) is a demonstrated endogenous substrate for caspase-3 in neurons undergoing unscheduled apoptotic death. We have previously identified the caspase cleavage site that yields the distinctive 120 kDa spectrin breakdown product (SBDP120) as (DSLD(1478)*SVEAL). Here, by using a synthetic peptide (NH(2)-SVEALC) mimicking the neo-N-terminal of SBDP120 as antigen, we report the development of chicken antibodies that specifically recognize the SBDP120 generated by in vitro caspase-3 digestion of bovine alpha-spectrin on Western blot. These anti-SBDP120 antibodies recognize SBDP120 generated by two apoptotic challenges (staurosporine, EGTA) to human neuroblastoma SH-SY5Y cells. Yet they neither react with intact alpha-spectrin nor its other fragments on Western blots. These anti-SBDP120 work equally well in detecting SBDP120 generated in rat cerebellar granule neurons undergoing potassium withdrawal induced apoptosis. In immunocytochemical studies, these antibodies also specifically stained apoptotic SH-SY5Y or CGN's undergoing apoptosis in a caspase inhibitor-sensitive manner. These anti-SBDP120s might become powerful markers for apoptotic neurons in various neurological or neurodegenerative conditions in vivo. PMID- 10825576 TI - Peripheral-type benzodiazepine receptors in human blood cells of patients affected by migraine without aura. AB - The kinetic parameters at equilibrium of peripheral benzodiazepine receptors in platelets, lymphocytes and granulocytes of 15 patients affected by migraine without aura were tested using [3H]PK 11195, a specific radioligand for this receptor and compared with the same number of healthy controls: a statistically significant increase (platelets 212%, lymphocytes 203%, granulocytes 171%, as absolute percentage) in the maximal number of binding sites (B(max)) in all three patient samples, compared with healthy controls was detected; on the contrary, the values of the dissociation constant (K(d)) at equilibrium do not show any statistically significant variations between the two groups. These data further confirm the presence of peripheral biochemical alterations in migraine without aura. As peripheral benzodiazepine receptors appear to be involved in the regulation of the mitochondrial respiratory chain, the observed increase in B(max) might be related to the mitochondrial anomalies found in migraine disorders. PMID- 10825577 TI - Alcohol enhances characteristic releases of dopamine and serotonin in the central nucleus of the amygdala. AB - The amygdaloid complex (AMY) is implicated in emotional and motivational aspects of behavior, including the formation of positive reinforcement association. AMY may also associated with brain rewarding circuitry. In the present study, the effect of ethanol (EtOH) on the release of dopamine (DA) and serotonin (5-HT) was studied in the central amygdaloid nucleus (CeAMY), and projecting excitatory afferents to the ventral tegmental area (VTA), of freely moving Wistar rats by brain microdialysis. Within 20 min of i.p. injection of EtOH (2 g/kg), the levels of DA and 5-HT in the CeAMY dialysate increased over the baseline value by 270 and 160% (N = 6-7), respectively. Addition of EtOH (25, 50 and 100 mM) to the microdialysis perfusion medium for 1 h caused a 115-150% dose-related increase in the extracellular level of DA in the CeAMY. 100 mM EtOH-induced CeAMY DA release continued to increase for 1 h after the perfusion medium was returned to normal perfusion medium. In contrast, the CeAMY 5-HT level was increased only by the addition of 100 mM EtOH for 1 h to 130% for 80 min. The stimulation of the CeAMY by EtOH through the microdialysis membrane showed delayed responses of DA and 5 HT compared with the i.p. injection of EtOH. Overall, the present findings are not sufficient to conclude whether EtOH acts directly or indirectly on the major monoamine nerve cells in the CeAMY, but the degree of acute EtOH action affected the differences in time at the peak response on EtOH-induced DA and 5-HT releases in the CeAMY via VTA. PMID- 10825578 TI - Effect of ionotropic glutamate receptors antagonists on the modifications in extracellular glutamate and aspartate levels during picrotoxin seizures: a microdialysis study in freely moving rats. AB - Our previous studies have shown a local decrease in glutamate and aspartate levels during seizures, induced by picrotoxin microdialysis in the hippocampus of chronic freely moving rats. In this paper, we study the effect of continuous hippocampal microperfusion of the NMDA, AMPA and kainate glutamate receptor inhibitors 5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5, 10-imine (MK 801); 6,7-dinitroquinoxaline-2,3-dione (DNQX), and 1-(4-aminophenyl)-4-methyl-7,8 methylenedioxy-5H-2,3-benzodiazepine hydrochloride (GYKI 52466). We also examine the action of L(-)-threo-3-hydroxyaspartic acid (THA), a glutamate and aspartate reuptake blocker, on the modification of extracellular glutamate and aspartate levels induced by picrotoxin, using the microdialysis method in freely moving rats. We found that changes in extracellular hippocampal concentrations in both amino acids are prevented by NMDA, AMPA and kainate receptor inhibitors. Seizures elicited under DNQX also induce a transient increase in aspartate extracellular levels coincident with seizure time. L(-)-threo-3-hydroxyaspartic acid increased the basal extracellular concentrations of both amino acids, but did not prevent the seizure-related decrease. Our results suggest that glutamate, the major neurotransmitter at the synaptic level, may also play an important role in non synaptic transmission during seizures. PMID- 10825579 TI - Distinction between Ca(2+) pump and Ca(2+)/H(+) antiport activities in synaptic vesicles of sheep brain cortex. AB - Synaptic vesicles, isolated from a sheep brain cortex, accumulate Ca(2+) in a manner that depends on the pH and pCa values. In the presence of 100 microM CaCl(2), most of the Ca(2+) taken up by the vesicles was vanadate-inhibited (86%) at pH 7.4, whereas at pH 8.5, part of the Ca(2+) accumulated (36%) was DeltapH dependent (bafilomycin and CCCP inhibited) and part was insensitive to those drugs (31%). We also observed that both vanadate-sensitive and bafilomycin sensitive Ca(2+) accumulations were completely released by the Ca(2+) ionophore, ionomycin, and that these processes work with high (K(0.5)=0.6 microM) and low (K(0.5)=217 microM) affinity for Ca(2+), respectively. The DeltapH-dependent Ca(2+) transport appears to be largely operative at Ca(2+) concentrations (>100 microM) which completely inhibited the vanadate-sensitive Ca(2+) uptake. These Ca(2+) effects on the Ca(2+) accumulation were well correlated with those observed on the vanadate-inhibited Ca(2+)-ATPase and bafilomycin-inhibited H(+) ATPase, respectively. The Ca(2+)-ATPase activity reached a maximum at about 25 microM (pH 7.4) and sharply declined at higher Ca(2+) concentrations. In contrast, Ca(2+) had a significant stimulatory effect on the H(+)-ATPase between 250 and 500 microM Ca(2+) concentration. Furthermore, we found that DeltapH sensitive Ca(2+) transport was associated with proton release from the vesicles. About 21% of the maximal proton gradient was dissipated by addition of 607.7 microM CaCl(2) to the reaction medium and, if CaCl(2) was present before the proton accumulation, lower pH gradients were reached. Both vanadate-inhibited and bafilomycin-inhibited systems transported Ca(2+) into the same vesicle pool of our preparation, suggesting that they belong to the same cellular compartment. These results indicate that synaptic vesicles of the sheep brain cortex contain two distinct mechanisms of Ca(2+) transport: a high Ca(2+) affinity, proton gradient-independent Ca(2+) pump that has an optimal activity at pH 7.4, and a low Ca(2+) affinity, proton gradient-dependent Ca(2+)/H(+) antiport that works maximally at pH 8.5. PMID- 10825580 TI - Cd4(+) T cell response to Leishmania spp. in non-infected individuals. AB - T cell mediated immunity is known to play a central role in the host response to control intra-cellular pathogens. This work demonstrates the presence of specific CD4(+) T cells to Leishmania spp. antigens in peripheral mononuclear cells of naive individuals (normal volunteers from non-endemic regions). The responder population was expanded by generation of antigen-specific T cell lines, which were produced by repeated stimulation with fixed promastigotes and autologous irradiated PBMC as antigen presenting cells. The leishmania-T cell lines were shown to proliferate in response to different species of the parasite (L. amazonensis, L. braziliensis, and L. donovani), but not to other recall antigens such as Candida albicans or tetanus toxoid. A preferential expansion of IFNgamma and IL-2 producing Th1-like T cells was observed. The leishmania-reactive cells were distributed between CD4(+) CD45RA(+) ("naive") and CD4(+) CD45R0(+) ("memory") populations. Although limiting dilution analysis showed a precursor frequency 3 times lower within the naive compartment, similar numbers of T cell lines were derived from both purified subpopulations. This study using leishmania specific CD4(+) T cell lines produced from normal individuals should provide information on cellular immune responses that are triggered by the parasite and how infection impacts the naive T cell repertoire. PMID- 10825581 TI - Bone marrow cells inhibit the generation of autologous EBV-specific CTL. AB - Recently, we reported that human bone marrow cells (BMC) inhibited the proliferative (recall) response of lymphocytes to Epstein-Barr virus (EBV) and cytomegalovirus (CMV) protein antigens [12]. To clarify further the effect of BMC on the immune response to viral antigens, we obtained PBL from EBV IgG antibody positive kidney transplant recipients (R) and their living-related donors (LRD) 1 year after renal transplantation and generated EBV-specific CTL in vitro in the presence or absence of autologous BMC. The addition of freshly aspirated autologous iliac crest BMC from either R or LRD caused a significant inhibitory effect on the generation of EBV-specific CTL from CTL precursors, in contrast to the addition of autologous PBL used as controls (62.29 +/- 10.85% inhibition using BMC from the kidney transplant recipients; 74.47 +/- 15.21% inhibition using BMC from the living-related donors). This inhibitory effect was only exerted during the CTL generation phase; but not in the effector CTL killing phase. The expression of CD94, a component of the killer inhibitory receptor (KIR) on CD3(+) cells was elevated in the cultures with BMC, in contrast to the cultures without BMC. The BMC inhibitory effect was partially abrogated by pre incubation of the CTL effectors with anti-CD94 monoclonal antibody, in contrast with its isotype control. In addition, supernatants obtained from the CTL generating cultures with BMC contained high levels of prostaglandin E(2) (PGE(2)), and EBV-specific CTL activity was inhibited by the addition of exogenous PGE(2) in the absence of BMC. The induction of CD40L cell surface expression by anti-CD3 was also decreased on the effector T cell population when BMC were added. There was a concomitant reduction in protein kinase C (PKC) activity. These studies demonstrate that BMC exert an inhibitory effect on T cell mediated immunity to viral antigens in humans by regulating autologous effector T cell generation and early T cell activation signaling pathways. PMID- 10825582 TI - Alloreactivity of umbilical cord blood mononuclear cells: specific hyporesponse to noninherited maternal antigens. AB - Earlier studies noted that patients who underwent cord blood (CB) transplantation had a lower incidence of graft-versus-host disease (GVHD) than those who underwent bone marrow transplantation (BMT). The premise that the immune reactivity of CB mononuclear cells (CB-MNC) to HLA mismatched combinations and to noninherited maternal antigens (NIMA) may be one of the factors involved in this phenomenon is still debatable. In this study we have attempted to evaluate the alloresponse and alloreactivity induced by CB-MNC by means of the standard mixed lymphocyte reaction test (SMLR) and the more sensitive, modified mixed lymphocyte reaction test (MMLR). Both techniques were used to test CB-MNC (n = 28) against HLA class II mismatched MNC from mothers (n = 26), fathers (n = 12), and unrelated individuals (n = 60) who served as controls. Alloresponse capabilities and stimulation capacities of CB-MNC in the SMLR were similar to those of control MNC: relative response (RR) = 73 vs. 65 and 58 vs. 65, respectively. Similar results were obtained in the MMLR. CB-MNC responded weakly to the maternal MNC in comparison with control MNC (RR = 47 vs. 73 [p = 0.0099]), while a stronger response was noted to the paternal than the maternal MNC (RR = 72 vs. 47 [p = 0.045]). Our results demonstrate that CB-MNC both respond to and induce alloresponse in HLA mismatched combinations. Moreover, the hyporesponse of CB-MNC to maternal cells that we observed suggests a form of tolerance to NIMA, which is probably due to the fetus's exposure to these antigens in its intrauterine life. PMID- 10825583 TI - Germ-free mice do not develop ankylosing enthesopathy, a spontaneous joint disease. AB - Ankylosing enthesopathy (ANKENT) is a naturally occurring joint disease in mice with numerous parallels to human ankylosing spondylitis (AS). Similarities between AS and ANKENT include not only affected tissue (joint entheses) but also association of the disease with genetic background, including MHC genes, gender, and age. Young males with the C57Bl/10 background have been described to suffer from ANKENT and, among H-2 congenic strains, high frequency of afflicted joints has been recorded in B10.BR (H-2(k)) males. Interestingly, the incidence of ANKENT is higher in conventional (CV) males that in their specific-pathogen-free (SPF) counterparts. The latter finding suggests that microbes could play a role as an ANKENT-triggering agent. To further examine this hypothesis we have established a germ-free (GF) colony of B10.BR mice and observed ANKENT incidence in both GF males and their conventionalized (ex-GF) male littermates; 20% of ex GF males developed ANKENT before 1 year of age. In contrast, no joint disease was observed under GF conditions (p < 0.0001). Our results show that live microflora is required in ANKENT pathogenesis. PMID- 10825584 TI - Soluble HLA levels in early pregnancy after in vitro fertilization. AB - Intact pregnancy can be interpreted as a state of maternal immunotolerance toward an haploidentical fetus. Soluble HLA (sHLA) molecules increase during episodes of allograft rejection and are discussed as candidates to modulate immune responses. We questioned whether after in vitro fertilization (IVF) the subsequent intact pregnancy, early abortion, or tubal pregnancy influence the courses sHLA serum levels. Therefore, serum samples of 65 IVF patients were assayed by ELISA for sHLA-I, sHLA-G, and sHLA-DR concentrations preovulatorily and after a positive HCG test weekly until the 9th gestational week (GW). In 20 patients experiencing an early abortion the preovulatory sHLA-G mean level of 25.9 +/- 3.9 SEM ng/ml and the share of 4.2 +/- 0.8 SEM % on total sHLA-I were significantly (p < 0.05) reduced compared to women with intact pregnancy. The same differences (p < 0.0001) were seen during the monitoring of sHLA-G and sHLA-I levels in intact pregnancy versus early abortion until 9th GW. Twin pregnancy revealed a drastically increase of sHLA-G levels from the 8th GW compared to singleton pregnancies. Further, individual sHLA-DR levels increased during intact pregnancy but decreased in the group of early abortion. With regard to sensitivity and specificity for pregnancy outcome sHLA quantitation reached similar weight as routine HCG determinations at GW 5. Especially women with preovulatory low sHLA-G levels appear to be on risk for early abortion after IVF. PMID- 10825585 TI - The generation of dendritic-like cells with increased allostimulatory function from acute myeloid leukemia cells of various FAB subclasses. AB - To increase the immunogenicity of leukemic cells, attempts were made to generate dendritic-like antigen presenting cells (DC) from AML blasts from 14 patients with AML FAB classifications M0-M5. Leukemic cells were cultured in the presence or absence of various cytokines including GM-CSF, SCF, TNF-alpha, IL-4, and gamma interferon. After various intervals recovery of viable cells was measured and expression of CD80, CD86, CD40, CD54, CD58, and CD11a was analyzed by flow cytometry. Functionally, DC derived from six AML samples were tested in a mixed lymphocyte response (MLR) using HLA-DR mismatched donor T cells as responder cells. Proliferation (5/14) or increased survival (7/14) of AML cells was observed in the presence of GM-CSF, SCF, and TNF-alpha. Only in the AML M2, M3, and M4 FAB subtypes proliferation was found. GM-CSF, SCF, and TNF-alpha induced morphologic changes typical for DC and increased the expression of costimulatory and adhesion molecules. No significant effect of IL-4 or gamma-interferon was observed. The day of maximal expression of these molecules varied. In cases with minor upregulation of CD80 or CD86, no further stimulation using CD40-L activation was observed. In the three cases tested, the DC-like cells retained the chromosomal abnormalities present in the original AML cells. In five out of six cases tested an increase in allostimulatory capacity was found at the day of maximal expression of costimulatory and adhesion molecules. In two patients a decrease in stimulatory capacity was found at day 7 compared with day 2 correlating with a decreased expression of these molecules. In conclusion, AML cells can be induced to increase their stimulatory capacity by upregulating costimulatory and adhesion molecules. PMID- 10825586 TI - Specific T cell deletion by transfected human monocytes expressing fas ligand and antigen. AB - The aim of our experiments was to determine whether deletion of antigen specific T helper cells could be accomplished by delivering the antigenic peptide to antigen presenting cells. Tetanus toxin peptide residues 830-843 was chosen for these experiments. Two mammalian expression vectors carrying the genes for human Fas ligand and a chimeric invariant chain-tetanus toxin peptide construct were designed. The T cell proliferative response to tetanus toxoid was inhibited when the antigen was presented by autologus monocytes transfected with Fas ligand. T cell mixture experiments using two syngeneic T cell lines specific either for tetanus toxoid or for pertussis toxin demonstrated that the killing effect elicited by the antigen pulsed/Fas ligand-transfected antigen presenting cells was antigen specific. Finally, we demonstrated that transient expression of antigen delivered by plasmid DNA can substitute for soluble antigen in the induction of antigen-specific T cell responses. Antigen presenting cells transfected with the vector carrying Fas ligand and the vector carrying the chimeric invariant chain-peptide antigen gene were shown to inhibit antigen specific T cell reactivity. This strategy may be useful for the induction of apoptosis in allopeptide reactive T cells driving chronic rejection. PMID- 10825587 TI - Human major histocompatibility molecules have the intrinsic ability to form homotypic associations. AB - We have investigated the homotypic associations of major histocompatibilty, class II and class I molecules using immunoprecipitation from detergent solubilised cell extracts. A 120-kDa structure corresponding to an HLA-DR dimer of dimers was immunoprecipitated by the HLA-DR specific mAb L243 from both biotinylated cell surface and metabolically labeled B cells and transfectant fibroblasts. The thermostability of this structure in SDS was examined. It was detected at 4 degrees C, 22 degrees C, and 37 degrees C, but not at 50 degrees C or 100 degrees C. Experiments performed with L243 Fab fragments and with purified HLA-DR molecules, indicated the presence of HLA-DR dimers of dimers and single heterodimers on B cells. HLA-DQ was also found to form SDS-stable dimers of dimers and single heterodimers on the cell surface of B cells, demonstrating that HLA class II isotypes, other than HLA-DR, also form homotypic associations. Similar experiments performed with HLA class I specific mAb, W632, revealed the existence of a 90 kDa and a 135-kDa structure corresponding to a MHC class I multimers. Under the same conditions, non-MHC molecules such as CD14 were found not to self-associate. These findings indicate that major histocompatibility molecules have the intrinsic ability to form homotypic associations at the cell surface of antigen presenting cells. PMID- 10825588 TI - Analysis of CIITA encoding AIR-1 gene promoters in insulin-dependent diabetes mellitus and rheumatoid arthritis patients from the northeast of Italy: absence of sequence variability. AB - Qualitative and/or quantitative alterations in the expression of the MHC class II molecules affect the onset and maintenance of the immune response and may be the basis of a wide variety of disease states, such as autoimmunity and immunodeficiency.CIITA is a major physiological regulator of the expression of MHC class II genes. The availability of CIITA ap- pears generally essential for MHC class II gene expression, and hence its own transcriptional regulatory mechanisms result of fundamental importance for a correct homeostasis of the immune response. Therefore, it is possible to hypothesize that variability at the CIITA-encoding locus, AIR-1, could constitute an additional source of susceptible traits to autoimmune diseases. Mutations at AIR-1/CIITA promoters could modulate expression of CIITA. Variations in CIITA expression could influence the qualitative and quantitative expression of MHC class II molecules at cell surface. We have analyzed sequence variation at AIR-1/CIITA promoters by PCR-SSCP in 23 IDDM and 30 RA patients compared to a sample of 19 unaffected normal controls and 16 unaffected IDDM family members, for a total of 88 Caucasian subjects from the Northeast of Italy. No sequence difference was found at the four AIR-1/CIITA promoters between autoimmune patients and normal controls. Moreover, the promoters resulted invariant within the entire group of 88 subjects analyzed, comprising patients and controls. This finding suggests a possible selective advantage in maintaining CIITA upstream regulatory sequences invariant. PMID- 10825589 TI - HLA-DRB1,3,4,5 and -DQB1 allele frequencies and HLA-DR/DQ linkage disequilibrium of 231 German caucasoid patients and their corresponding 821 potential unrelated stem cell transplants. AB - Allelic matching within the HLA-DRB1 and -DQB1 loci significantly improves the clinical outcome of hematopoietic stem cell transplantation. Consequently, allelic typing of these loci is strongly recommended for the unrelated stem cell donor selection. In this study, the HLA-DRB1,3,4,5 and -DQB1 alleles of 231 patients and their corresponding 821 nonrandom potential stem cell donors were determined to define compatible donor/recipient pairs. Highly accurate HLA typing data were achieved by PCR-SSOP and a combination of group specific PCR-SSP and subsequent sequencing-based typing of nearly the whole second exon of each locus. The alleles DRB1*07, *09, and *10 were analyzed by PCR-reverse dot blot hybridization instead of sequencing. Additionally, DRB1 homozygosity was verified by temperature gradient gel electrophoresis. The identified 2104 HLA-DRB1 and HLA DQB1 alleles as well as data on HLA-DRB3, -DRB4, and -DRB5 alleles were applied to a statistical program and absolute and relative delta values of DR/DQ linkages were calculated. The achieved data on the HLA-DRB1 allele distribution and on DR/DQ associations in terms of subtypes significantly ensure the typing reliability, since rare allele combinations will result in further investigations. Furthermore, detailed data on the DR/DQ allele associations allow estimations of the number of HLA-A, -B, and -DR matched unrelated stem cell donors necessary for the identification of DRB and DQB subtype identical donors. PMID- 10825590 TI - MHC class I and class II phenotype, gene, and haplotype frequencies in Greeks using molecular typing data. AB - In the present study, DNA typing for HLA-A, C, B, DRB1, DRB3, DRB4, DRB5, DQA1, DQB1, and DPB1 was performed for 246 healthy, unrelated Greek volunteers of 20-59 years of age. Phenotype, genotype frequencies, Hardy-Weinberg equilibrium fit, and 3-locus haplotype frequencies for HLA-A, C, B, HLA-A, B, DRB1, HLA-DRB1, DQA1, DQB1, and HLA-DRB1, DQB1, DPB1 were calculated. Furthermore, linkage disequilibrium, deltas, relative deltas and p-values for significance of the deltas were defined. The population studied is in Hardy-Weinberg equilibrium, and many MHC haplotypes are in linkage disequilibrium. The most frequent specificities were HLA-A*02 (phenotype frequency = 44.3%) followed by HLA-A*24 (27.2%), HLA-B*51 (28.5%), HLA-B*18 (26.8%) and HLA-B*35 (26.4%) and HLA-Cw*04 (30.1%) and HLA-Cw*12 (26.8%). The most frequent MHC class II alleles were HLA DRB1*1104 (34.1%), HLA-DQB1*0301 (54.5%) and HLA-DPB1*0401 with a phenotype frequency of 59.8%. The most prominent HLA-A, C, B haplotypes were HLA-A*24, Cw*04, B*35, and HLA-A*02, Cw*04, B*35, each of them observed in 21/246 individuals. The most frequent HLA-A, B, DRB1 haplotype was HLA-A*02, B*18, DRB1*1104 seen in 20/246 individuals, while the haplotype HLA-DRB1*1104, DQB1*0301, DPB1*0401 was found in 49/246 individuals. Finally, the haplotype DRB1*1104, DQA1*0501, DQB1*0301 was observed in 83/246 individuals. These results can be used for the estimation of the probability of finding a suitable haplotypically identical related or unrelated stem cell donor for patients of Greek ancestry. In addition, they can be used for HLA and disease association studies, genetic distance studies in the Balkan and Mediterranean area, paternity cases, and matching probability calculations for the optimal allocation of kidneys in Greece. PMID- 10825591 TI - Age-related association of MHC class I chain-related gene A (MICA) with type 1 (insulin-dependent) diabetes mellitus. AB - To assess the contribution of the HLA class I region to susceptibility to and heterogeneity of type 1 diabetes, we investigated the association of polymorphism of MHC class I chain-related gene A (MICA) with age-at-onset as well as susceptibility to type 1 diabetes. One hundred one Japanese patients and 110 healthy control subjects were studied. The frequency of A4 allele was significantly higher and that of A6 allele was significantly lower in patients than in control subjects. The frequency of A5.1 allele was highest in early-onset patients (23.0%), intermediate in intermediate-onset patients (9.2%) and lowest in late-onset patients (7.7%) (trend chi-squared test, p = 0.0098). A5. 1 allele was strongly associated with HLA-B7 and Cw7, suggesting that MICA*A5.1-B7-Cw7 haplotype contains a gene responsible for age-at-onset. A4 allele was associated with a susceptible haplotype, DR4-DQB1*0401, and A6 allele was associated with a protective haplotype, DR2-DQB1*0601, suggesting that the association of MICA with type 1 diabetes susceptibility may be due to linkage disequilibrium with class II haplotypes. These data suggest that MICA gene is associated with age-at-onset and that a gene (or genes) responsible for age-at-onset of type 1 diabetes is located in the HLA class I region, probably near the region of MICA-B-C. PMID- 10825592 TI - Nomenclature for factors of the HLA system, update February 2000. WHO Nomenclature Committee for Factors of the HLA System. PMID- 10825593 TI - DNA vaccine encoding nucleocapsid and surface proteins of wild type canine distemper virus protects its natural host against distemper. AB - Canine distemper virus (CDV), a member of the genus Morbillivirus induces a highly infectious, frequently lethal disease in dogs and other carnivores. Current vaccines against canine distemper consisting of attenuated viruses have been in use for many years and have greatly reduced the incidence of distemper in the dog population. However, certain strains may not guarantee adequate protection and others can induce post vaccinal encephalitis. We tested a DNA vaccine for its ability to protect dogs, the natural host of CDV, against distemper. We constructed plasmids containing the nucleocapsid, the fusion, and the attachment protein genes of a virulent canine distemper virus strain. Mice inoculated with these plasmids developed humoral and cellular immune responses against CDV antigens. Dogs immunized with the expression plasmids developed virus neutralizing antibodies. Significantly, vaccinated dogs were protected against challenge with virulent CDV, whereas unvaccinated animals succumbed to distemper. PMID- 10825594 TI - Immunization of rabbits with cottontail rabbit papillomavirus E1 and E2 genes: protective immunity induced by gene gun-mediated intracutaneous delivery but not by intramuscular injection. AB - We previously demonstrated that gene gun-based intracutaneous vaccination of rabbits with a combination of, but not with individual papillomavirus E1, E2, E6 and E7 genes provided complete protection against cottontail rabbit papillomavirus (CRPV) infection. In the present study, we tested whether vaccination of inbred and outbred rabbits with a combination of CRPV E1 and E2 genes could provide complete protection against virus infection. In the first experiment, gene gun-based intracutaneous vaccination with E1 and E2 genes prevented papilloma formation in the majority of inbred rabbits and promoted systemic papilloma regression in one non-protected rabbit. In contrast, needle mediated intramuscular injection of E1 and E2 genes did not prevent papilloma formation nor promoted systemic papilloma regression, indicating an absence of strong protective immunity. In the second experiment, six outbred rabbits were immunized by gene gun-based intracutaneous administration of the E1 and E2 genes. Prevention of papilloma formation or systemic papilloma regression was observed in three vaccinated rabbits. Papillomas persisted on the remaining three rabbits, but were significantly smaller than that on control rabbits. These results suggested that gene gun-based intracutaneous vaccination with the combination of papillomavirus E1 and E2 genes induced strong protective antivirus immunity but may be insufficient for complete protection in an outbred population. PMID- 10825596 TI - The incidence of vaccine preventable influenza-like illness and medication use among Pakistani pilgrims to the Haj in Saudi Arabia. AB - Over the 33-day duration of the 1999 Haj in Saudi Arabia, we collected daily health status reports for 2070 Pakistani pilgrims over 13 years of age, 54% of whom had elected to receive influenza vaccine immediately before departing for the Haj. We calculated vaccine preventable outcome incidence as the difference in attack rates between vaccinated and unvaccinated persons. The incidences of vaccine preventable influenza-like illness (sore throat in combination with cough or fever of at least 38 degrees C), fever, and any symptom of upper respiratory infection were 22, 17, and 24 per 100 pilgrims per Haj. For every 100 persons who attended the Haj, 17 had a course of antibiotics and 23 had a course of nonprescription cold medication that was preventable with influenza vaccine use. Influenza leads to significant morbidity and medication use among Haj pilgrims. Vaccine against influenza should be considered for pilgrims before entry into Saudi Arabia. PMID- 10825595 TI - Intramuscular immunization with genetically inactivated (ghosts) Actinobacillus pleuropneumoniae serotype 9 protects pigs against homologous aerosol challenge and prevents carrier state. AB - Bacterial ghosts are empty cell envelopes achieved by the expression of a cloned bacteriophage lysis gene and, unlike classical bacterins, suffer no denaturing steps during their production. These properties may lead to a superior presentation of surface antigens to the immune system. Currently available porcine Actinobacillus pleuropneumoniae vaccines afford only minimal protection by decreasing mortality but not morbidity. Pigs which survive infection can still be carriers of the pathogen, so a herd once infected remains infected. Carrier pigs harbour A. pleuropneumoniae in their nasal cavities, in their tonsils, or within lung lesions. A dose-defined nose-only aerosol infection model for pigs was used to study the immunogenic and protective potential of systemic immunization with ghosts made from A. pleuropneumoniae serotype 9 reference strain CVI 13261 against an homologous aerogenous challenge. Pigs were vaccinated twice intramuscularly with a dose of 5x10(9) CFU ghosts (GVPs) or formalin inactivated A. pleuropneumoniae bacterins (BVPs). After 2 weeks vaccinated pigs and non-vaccinated placebo controls (PCs) were challenged with a dose of 10(9) CFU by aerosol. The protective efficacy of immunization was evaluated by clinical, bacteriological, serological and post-mortem examinations. Bronchoalveolar lavage in pigs was performed during the experiment to obtain lavage samples (BALF) for assessment of local antibodies. Isotype-specific antibody responses in serum and BALF were determined by ELISAs based on whole cell antigen. Immunization with ghosts did not cause clinical side-effects. After aerosol challenge PCs developed fever and pleuropneumonia. GVPs or BVPs were found to be fully protected against clinical disease or lung lesions in both vaccination groups, whereas colonization of the respiratory tract with A. pleuropneumoniae was only prevented in GVPs. Specific immunoglobins against A. pleuropneumoniae were not detectable in BALF after immunization. A significant systemic increase of IgM, IgA, IgG(Fc'), or IgG(H+L) antibodies reactive with A. pleuropneumoniae was measured in GVPs and BVPs when compared to the non-exposed controls. BVPs reached higher titers of IgG(Fc') and IgG(H+L) than GVPs. However, prevention of carrier state in GVPs coincided with a significant increase of serum IgA when compared to BVPs. These results suggest that immunization with ghosts, that bias antibody populations specific to non-denaturated surface antigens, may be more efficacious in protecting pigs against colonization and infection than bacterins. PMID- 10825597 TI - Adverse events after Japanese encephalitis vaccination: review of post-marketing surveillance data from Japan and the United States. The VAERS Working Group. AB - We determined the reporting rates for adverse events following the administration of inactivated mouse-brain derived Japanese encephalitis vaccine (JEV) based on post-marketing surveillance data from Japan and the United States. The rate of total adverse events per 100,000 doses was 2.8 in Japan and 15.0 in the United States. In Japan, 17 neurological disorders were reported from April 1996 to October 1998 for a rate of 0.2 per 100,000 doses. In the United States, no serious neurological adverse events temporally associated with JEV were reported from January 1993 to June 1999. Rates for systemic hypersensitivity reactions were 0.8 and 6.3 per 100,000 doses in Japan and the United States, respectively. Passively collected VAERS surveillance data indicate that characteristic hypersensitivity reactions with a delayed onset continue to occur among JEV recipients and that conservative recommendations limiting its use to travelers at high risk of infection with Japanese encephalitis are appropriate. PMID- 10825599 TI - Adequate immune response to tetanus toxoid and failure of vitamin A and E supplementation to enhance antibody response in healthy children. AB - The effects of vitamin A and vitamin E supplementation on the IgG response to tetanus toxoid after primary immunization were evaluated in a prospective, randomized controlled clinical trial involving 89 healthy infants with normal serum vitamin A and E levels at 2 months of age. Before the first dose of DPT vaccine, the infants were randomly enrolled into four different study groups [Group I (n=24): 30,000 IU vitamin A for 3 days just after each three doses of primary vaccination, Group II (n=21): 150 mg oral vitamin E for only 1 day after the injections for primary immunization, Group III (n=21): vitamins A and E together in the same order, Group IV (n=23) no vitamin after DPT vaccines]. Serum tetanus antitoxin (IgG) titres were measured three times; initially at 2 months of age before the first dose of DPT, secondly at 5 months of age 1 month after primary immunization and thirdly at 16-18 months of age before the booster dose of DPT. Before the first dose of the DPT vaccine, 1 month after the third DPT injection and at 16-18 months before the booster dose of DPT, there was no significant difference in serum tetanus antitoxin levels between these four groups. A significant increase was observed in all the groups when serum tetanus antitoxin levels before (2 months) and after (5 months) primary immunization were compared. In addition, serum antibody levels against tetanus significantly decreased in the four groups before booster vaccination. Before the beginning of primary immunization, 15 infants (16.8%) had serum tetanus antitoxins (IgG) below protective level. After three doses of DPT, all the infants had protective antitoxin levels. At 16-18 months of age before booster dose, four infants (10%) also had serum tetanus antitoxins (IgG) below the protective level. No side effects were observed except bulging fontanelle in two infants in Group I. PMID- 10825598 TI - Failure of female baboons (Papio anubis) to conceive following immunization with recombinant non-human primate zona pellucida glycoprotein-B expressed in Escherichia coli. AB - Progress in the development of an immunocontraceptive vaccine based on zona pellucida glycoproteins has been hampered due to observed ovarian dysfunction associated with immunization using these as immunogens. In this study four female baboons (Papio anubis) were immunized with recombinant bonnet monkey (Macaca radiata) zona pellucida glycoprotein-B (r-bmZPB) expressed in Escherichia coli and conjugated to diphtheria toxoid (DT) using Arlacel-A and Squalene as adjuvants. All the immunized animals elicited a good antibody response against r bmZPB, continued to have ovulatory cycles and showed no disturbance in the cyclicity. In presence of high titres of circulating anti-bmZPB antibodies (>2x10(3) antibody units), the immunized animals failed to conceive following mating with males of proven fertility. Pregnancy was observed in the immunized animals subsequent to the decline in anti-r-bmZPB antibody titres. These results, though preliminary, suggest that immunization with ZPB may be used for immunocontraception without obvious ovarian dysfunction. PMID- 10825600 TI - Successful DNA immunisation of rats against fasciolosis. AB - The liver fluke Fasciola hepatica contributes to great economic and health losses in the cattle industry in many countries, including Poland. Unfortunately, no vaccine against fasciolosis is commercially available. We have designed a DNA vaccine and tested it in rats. Groups of male or female rats received one intramuscular injection of 50 microg of a pcDNA 3.1 vector carrying cDNA encoding for a cysteine proteinase of F. hepatica. The plasmid was diluted in saline containing 0.05% bupivacaine. Control rats were injected with empty plasmid or not injected at all. All rats were challenged with 45 metacercariae of the fluke on day 28 of the experiment. Seven weeks after the challenge infection fluke burdens were evaluated in vaccinated and control rats. Male rats vaccinated with cysteine proteinase cDNA revealed 100% protection against F. hepatica infection. Females immunised in the same way exhibited the reduction of fluke burden by 74%. PMID- 10825601 TI - Intranasal immunization with protein-linked phosphorylcholine protects mice against a lethal intranasal challenge with streptococcus pneumoniae. AB - Immunization against phosphorylcholine (PC) linked to a protein protects mice against Streptococcus pneumoniae when used parenterally, and against Salmonella typhimurium when used orally after entrapment in D,L-Lactide-co-Glycolide microspheres. Here, we immunized BALB/c mice intranasally with a serotype 3 S. pneumoniae strain. Immunization was followed by a rise in anti-PC IgA and IgG titers in serum and in pulmonary secretions, but not by any rise in anti ds-DNA antibody nor any glomerular Ig deposition. The survival rates were 91 and 76% in the two groups of mice, respectively. These rates were significantly higher than those in control mice immunized intranasally either with Thyr loaded in microspheres (0%), blank microspheres (22%), free Thyr (17%), and saline (18%). This demonstrates that the mucosal route is effective for vaccination against S. pneumoniae pneumonia with PC linked to a protein carrier. It constitutes another important step forward in the development of the concept that PC can be used as a mucosal immunogen for protection against the different diseases caused by PC bearing bacteria. PMID- 10825602 TI - Humoral immunity against diphtheria, tetanus and poliomyelitis after antineoplastic therapy in children and adolescents--a retrospective analysis. AB - Serum antibodies against diphtheria- and tetanus-toxin were measured in 71 children and against poliomyelitis viruses 1-3 in 65 children and adolescents 0 18 months after cessation of antineoplastic therapy. Non or marginally protective serum titers were found in 62% of patients against diphtheria, in 18% of patients against tetanus and in 72% of patients against one or more poliomyelitis virus serotypes. Of these patients, 55%, 46% and 75% were immunized adequately according to age against diphtheria, tetanus and poliomyelitis, respectively. In 50% or more of patients a lack of protective immunity against diphtheria, tetanus and poliomyelitis was found which could not be explained by an inadequate immunization status. This suggests that other factors (e.g. influence of underlying illness, antineoplastic therapy or both on lymphocytes) might be responsible for these findings and this deserves further investigation. Measurement of serum antibodies against vaccine-preventable illnesses and consecutive booster immunizations are an essential part of long-term follow up in pediatric patients after antineoplastic therapy. PMID- 10825603 TI - Active versus passive immunization against hepatitis A in the Israel defence forces: a cost-benefit analysis. AB - We compared cost-benefit and cost-effectiveness analyses of hepatitis A prevention with immune serum globulin (ISG) vs inactivated hepatitis A vaccine (iHAV) in the Israel Defence Forces. Personnel were grouped according to conditions and duration of service and analyses were performed based on maximum and minimum hepatitis A projected incidences for each group. For standing army soldiers in field units, iHAV is economically superior compared with ISG. For the five other groups studied, the reverse is true. Expected increases in production costs of ISG and expected decreases in costs of iHAV are likely to make iHAV more economical in the future. PMID- 10825604 TI - Therapy of murine cutaneous leishmaniasis by DNA vaccination. AB - Prophylactic DNA vaccination protects mice against infection with Leishmania major by inducing an exclusive Th1 immune response dominated by the production of IFN-gamma. Here we show that DNA vaccines, initially designed to prevent infection, can also have a significant therapeutic effect. In L. major infected mice, vaccination with DNA encoding the Parasite Surface Antigen/gp46/M2 causes reduction in lesion size and promotes healing in both genetically resistant C3H/He mice and susceptible BALB/c mice. The therapeutic effect is underpinned by a shift in the T cell-derived cytokine environment with an increase in the IFN gamma producing Th1 type cells. Application of such immunotherapy in conjunction with antiparasite drugs may result in faster or more certain cure of the disease in humans. PMID- 10825605 TI - Absence of shedding of two B. abortus strains in goats after vaccination with live vaccines. AB - The objective of this study was to determine if B. abortus rough mutant strains RB51 and rfbK are eliminated in goat milk. Thirty milk goats were divided into two groups. Group I was inoculated with 4x10(10) cfu/ml of B.abortus RB51 strain and Group II with 1x10(9) cfu/ml of B. abortus rfbK strain by subcutaneous route in the right axilary region. Milk samples were taken aseptically on a daily basis for the first 30 days and weekly for the second and third months. The samples were inoculated on Farrell's media and incubated under 5-10% CO(2) at 37 degrees C for 10 days. The suspicious colonies were recultured in Farrell's media. There were no isolations of bacteria with characteristics of Brucella from any of the milk samples collected during 90 days of the study. It was concluded that neither of the strains used at these doses were eliminated by milk in goats inoculated during lactation. PMID- 10825606 TI - Complex cytokine responses to hepatitis B surface antigen and tetanus toxoid in responders, nonresponders and subjects naive to hepatitis B surface antigen. AB - Some human subjects vaccinated with hepatitis B surface antigen (HBsAg) do not produce antibodies to the vaccine (nonresponders). The mechanism for nonresponse is unknown. To understand the response and nonresponse to nominal antigens better, we determined the level and kinetics of cytokine secretion in response to HBsAg and tetanus toxoid (TT) by peripheral blood mononuclear cells (PBMC) in vitro from HBsAg vaccine responders and nonresponders and from individuals naive to HBsAg. Proliferating PBMC secreted peak levels of interleukin-2 (IL-2) at 2 days and peak levels of tumor necrosis factor-beta (TNF-beta), interferon-gamma (IFN-gamma), IL-4 and IL-10 at 3-6 days post-stimulation. In contrast, nonproliferating PBMC (whether from nonresponders, naive subjects or weak responders) did not produce detectable levels of TNF-beta or IFN-gamma, nor was IL-4 or IL-10 produced significantly, and that produced had a different kinetic profile from that of proliferating PBMC. HBsAg-specific cytokine production by PBMC from strong responders broadly paralleled their cytokine responses to TT. Cellular cytokine mRNA levels measured by reverse transcriptase-polymerase chain reaction corroborated the secreted cytokine results. The anti-HBsAg- and anti-TT specific T cell cytokine responses were mixed Th(1/2)-like and donor-specific. An HBsAg-specific cytokine response, but not a TT-specific cytokine response, was completely missing in nonresponders. These data suggest that the T cell defect of HBsAg nonresponse is not due to a skewed cytokine profile. PMID- 10825607 TI - Local and systemic immune responses to combined vibrio cholerae CVD103-HgR and salmonella typhi ty21a live oral vaccines after primary immunization and reimmunization. AB - The local and systemic antibody responses elicited following concomitant primary immunization and reimmunization with the live oral attenuated Vibrio cholerae CVD103-HgR and Salmonella typhi Ty21a vaccine strains were determined in healthy adult volunteers. A more pronounced serum vibriocidal antibody response was generated after primary immunization compared to reimmunization 2.5 or 3.5 yr later. The seroconversion rate (> or =4-fold rise over baseline) was 81% subsequent to primary immunization versus 57% (p=0.018) and 65% (p=0.639) upon reimmunization at 2.5 and 3.5 yr, respectively. A similar trend was observed for serum anti-S. typhi lipopolysaccharide (LPS) antibodies. After primary immunization, 48% of subjects manifested a significant rise in coproantibody levels to V. cholerae LPS while 60% did so for cholera toxin (CT). Upon reimmunization, the response rate for LPS ranged from 38% at 2.5 yr to 56% at 3.5 yr (p>0.05), while that for CT varied from 31% (p=0. 007) to 50% (p=0.541) at 2.5 and 3.5 yr, respectively. The anti-S. typhi IgA coproantibody response rate was 70% subsequent to primary immunization versus 47% at 2.5 yr (p=0.021) and 63% at 3.5 yr (p=0. 77). PMID- 10825608 TI - Antibody response after influenza vaccination in HIV-infected individuals: a consecutive 3-year study. AB - In a consecutive 3-year study the antibody response after immunization with influenza vaccine of a cohort of HIV-infected adults was studied. The haemagglutination-inhibiting (HAI) antibody titres after vaccination correlated with the number of CD4(+) T lymphocytes (p<0.001), the prevaccination antibody titres (p<0.001), and the proliferative response to anti-CD3 (p<0.001). Severely impaired antibody responses were observed in HIV-infected individuals with CD4(+) T-lymphocyte counts < or =100x10(6)/l. Significantly higher prevaccination antibody titres were observed in healthy controls in the 2nd or 3rd year of vaccination, but not in HIV-infected individuals. Annually repeated vaccination of HIV-infected individuals did not lead to higher postvaccination antibody titres. Annual vaccination of HIV-infected individuals with CD4(+) T-lymphocyte counts exceeding 100x10(6)/l seems to be worthwhile, although it may not be expected to render the same level of protection against influenza as in non infected individuals. PMID- 10825609 TI - DNA-based vaccination against murine paracoccidioidomycosis using the gp43 gene from paracoccidioides brasiliensis. AB - Gp43, the major 43-kDa antigenic glycoprotein of Paracoccidioides brasiliensis, or its 15-amino acid inner peptide (P10), induces a T-CD4(+), Th1 cellular immune response which protects BALB/c mice from intratracheal infection by virulent yeast forms. We investigated whether DNA vaccination using the gp43 gene could elicit protective immunity against P. brasiliensis. Animals immunised intramuscularly (i.m.) or intradermally (i.d.) with plasmid DNA containing the gp43 gene induced a specific, long lasting humoral and cellular immune response. A mixed Th1/Th2 cellular immune response in DNA-immunized mice was modulated in vivo by IFN-gamma and was protective in BALB/c mice. A significant decrease in the lung colony forming units (CFUs) and reduced, or no dissemination to the spleen and liver of immunised mice were observed. PMID- 10825610 TI - The effectiveness of the infant hepatitis B immunisation program in Fiji, Kiribati, Tonga and Vanuatu. AB - The aims of this project were: (1) to determine the extent to which infant hepatitis B immunisation is preventing chronic hepatitis B infection in children living in a sample of Pacific Island countries; and (2) to identify factors associated with the successful prevention of hepatitis B infection in these populations. A regional hepatitis B immunisation project which supplied hepatitis B vaccine to 10 Pacific Island countries began in 1995. Seroepidemiological surveys were conducted in Fiji, Kiribati, Tonga and Vanuatu in early 1998. These included immunised pre-school children and their biological mothers, and a historical control group of unimmunised students. Prevalence rates for hepatitis B surface antigen (HBsAg) in the populations of students, mothers and their pre school children were respectively: Fiji: 6.9, 6.6, 0.7%; Kiribati: 27.4, 15.1, 3.8%; Tonga: 11.1, 18.6, 3.8%; Vanuatu: 16.3, 12.3, 3.0%; and for all four countries: 13.2, 12.5, 2.6%. Compared to the historical control group of students, the pre-school population had a much lower probability of HBsAg positivity (relative risk [RR]=0.19 [95%CI: 0.12-0.31]). Statistically significant differences in risk were apparent for all the countries: Fiji: RR=0.10; Kiribati: RR=0.14; Tonga: RR=0.34; Vanuatu: RR=0.19. This is equivalent to an overall program effectiveness of 81% (95%CI: 69-88%) in reducing chronic carriage. Also, the overall protective effectiveness against vertical hepatitis B transmission resulting in HBsAg positivity among children exposed to HBeAg positive and negative carrier mothers, was estimated to be 70%. By age 6 months, when all children should have had three vaccine doses, completed immunisation rates ranged from 22 (Fiji) to 84% (Vanuatu). Coverage of the first dose being given within 2 days of birth varied from 43% in Kiribati to 92% in Tonga. In conclusion hepatitis B immunisation of infants in these four countries is having a substantial beneficial effect in preventing chronic hepatitis B infection. Nevertheless, there is significant scope for further improving the timeliness of immunisation. PMID- 10825611 TI - Improved mucosal protection against Venezuelan equine encephalitis virus is induced by the molecularly defined, live-attenuated V3526 vaccine candidate. AB - The genetically engineered, live-attenuated Venezuelan equine encephalitis (VEE) virus vaccine candidate, V3526, was evaluated as a replacement for the TC-83 virus vaccine. Protection from lethal subcutaneous or aerosol challenge was evaluated in vaccinated mice clinically and immunohistochemically. Subcutaneous administration of V3526 induced systemic and mucosal protection more efficiently than did the TC-83 vaccine. The bronchial IgA responses induced in mice by subcutaneous administration of vaccines significantly corresponded to the ability to survive aerosol challenge with virulent virus. Furthermore, V3526 delivered by aerosol induced more complete mucosal protection than either vaccine administered subcutaneously. The ability of V3526 to induce protection in mice warrants its consideration for further testing as a potential vaccine candidate for human use. PMID- 10825612 TI - In this issue PMID- 10825613 TI - The resuscitation greats. Benjamin Pugh: the air-pipe and neonatal resuscitation. PMID- 10825614 TI - Resuscitation in shock associated with burns. Tradition or evidence-based medicine? AB - OBJECTIVE: To summarize the present standards and guidelines for fluid treatment of shock associated with burns, and to evaluate their scientific support in the literature. DESIGN: Nonsystematic, critical review of the literature regarding the indications for crystalloid and colloid fluid treatment, invasive monitoring and the use of resuscitation end points in shock associated with burns. SUMMARY POINTS: Crystalloid fluid resuscitation of patients with burns is traditionally managed using empirical resuscitation formulae, with the efficacy monitored by vital signs and urinary output The value of these end points has been questioned by recent studies, which have suggested that such noninvasive parameters may be inadequate for detecting malperfusion. No consensus exists regarding appropriate assessment of adequate resuscitation, and the impact on survival of invasive measures has still to be proven in controlled randomized trials. Generally, a significantly higher fluid requirement has been demonstrated when resuscitation is based on invasive cardiorespiratory monitoring. Colloid resuscitation in burns patients is controversial. Published reports suggest that colloid infusion should be started between 6 and 36 h following thermal injury. A recent meta-analysis highlighted the shortcomings of albumin in patients with burns, and this, together with restrictions for the use of plasma products, has obscured the choice of colloid solution. The effect of colloid resuscitation on survival remains to be proven in burned patients. CONCLUSION: The current standards for monitoring fluid therapy in patients with large burns are not supported by scientific data. Further randomized, controlled trials are indicated, and should help establish general guidelines regarding monitoring and treatment end points in these patients. PMID- 10825615 TI - Early defibrillation and the chain of survival in 'in-hospital' adult cardiac arrest; minutes count. AB - OBJECTIVE: To report the outcomes from and the impact of the chain of survival in 'in-hospital' cardiac arrest where the presenting rhythm was VF/VT, the arrest was witnessed, defibrillation was conducted rapidly and no other resuscitation interventions were required. OUTCOME MEASURES: Any return of spontaneous circulation and discharge from hospital. METHODS: A 2-year prospective resuscitation audit using the Utstein style was conducted within a major London NHS Hospital Group. RESULTS: There were 124 patients who had primary VF/VT arrest. Eight were excluded from the study and 14 had non-witnessed cardiac arrest. Twenty one patients had witnessed VF/VT arrest but with delayed defibrillation, 81 patients had witnessed VF/VT arrest with rapid defibrillation, 69 patients had witnessed VF/VT arrest with rapid defibrillation, CPR and other additional interventions. There were 15 patients that had witnessed cardiac arrest with a presenting rhythm of VF/VT, who received rapid defibrillation and had no ventilation or chest compression prior to or following defibrillation. All 15 patients achieved a return of spontaneous circulation, and 12 were discharged alive. CONCLUSIONS: Rapid defibrillation prior to any other resuscitation intervention is associated with increased survival from witnessed VF/VT arrest in in-hospital cardiac arrest victims, and that the time to first shock is critical in enhancing the prospects of long-term survival in these patients. PMID- 10825616 TI - An audit of basic life support skills amongst general practitioner principals: is there a need for regular training? AB - OBJECTIVE: to determine what proportion of general practitioner (GP) principals in Herefordshire health authority are able to perform basic life support according to current guidelines. DESIGN: prospective cohort study. SETTING: Herefordshire Health Authority area. SUBJECTS: 53 general practitioner (GP) principals, based in 18 surgeries. MAIN OUTCOME MEASURES: basic life support (BLS) skills were assessed using the 1997 UK Resuscitation Council guidelines in checklist form as a process criterion. An assessments of each doctor was made before and after teaching of BLS skills. RESULTS: 50% of GP principals (n=53) in Herefordshire Health Authority took part in the audit. Of these 90.6% (48) were unable to perform BLS according to current guidelines on initial appraisal. After a practical teaching session 98% were competent in BLS skills. CONCLUSION: without regular teaching, BLS skills amongst GPs are likely to fall below an acceptable standard. Even short teaching periods can produce significant improvement in skills. PMID- 10825617 TI - The seeing stylet: a new device for tracheal intubation. AB - A new tracheal intubation device is available. The 'Shikani Seeing Stylet' is a new, inexpensive, reusable high resolution endoscope with a malleable stainless steel sheath which can be inserted through a tracheal tube allowing intubation to be performed under direct vision. We have assessed this new device on 20 patients (ASA I-II; age 25-67) scheduled to undergo elective surgery with tracheal intubation. We measured heart rate (HR), non invasive blood pressure (NIBP), oxygen saturation (SpO(2)) and end tidal carbon dioxide (ETCO(2)) at three different times: T(0) (induction of anesthesia), T(1) (beginning of intubation procedure), T(2) (end of intubation procedure); we also recorded the time interval between T(1) and T(2). All patients were successfully intubated with the device. Eleven patients were intubated at the first attempt (T(1)-T(2) mean time=8. 65 s); three patients were intubated at the first attempt using cricoid pressure (T(1)-T(2) mean time 11.6 s); four patients were intubated at the second attempt (T(1)-T(2) mean time=36.5 s); two patients were intubated at the third attempt (T(1)-T(2) mean time=54. 5 s). The HR, NIBP, SpO(2) and ETCO(2) remained fairly stable. On the basis of our preliminary experience with 20 patients, the 'Shikani Seeing Stylet' seems to be a promising adjunct for airway management. PMID- 10825618 TI - COPA as an aid for tracheal intubation. AB - The purpose of this study is to verify the usefulness of the cuffed oropharyngeal airway (COPA) as a device to guide a tracheal tube using a semiblind technique with a lightwand. Ten anaesthetised patients (ASA I-II, aged 35-67) undergoing to an elective surgery were analysed. We selected and positioned a correct size of COPA for each patient. A lightwand (Trachlight) was then inserted into the COPA to confirm correct placement of this device. The lightwand was then removed and the first portion of a tube exchanger (TE) was inserted and connected by a 15-mm connector with the breathing circuit and its position was confirmed by End Tidal CO(2) values during ventilation. The patients were then paralysed and ventilation through the first portion of the TE reconfirmed. The COPA was removed, and the second portion of the TE was connected and used as a guide for a tracheal intubation. This combined technique had a success rate of six out of ten patients and could be used for airway management if a fibre optic scope or other devices such as a Combitube, LMA or LMA Fastrach were not available. The preliminary data from this study are not indicative of the statistical validity of this technique. Further studies should be performed to verify the statistical reliability of the technique. PMID- 10825619 TI - Obstacles to bystander cardiopulmonary resuscitation in Japan. AB - OBJECTIVE: bystander cardiopulmonary resuscitation (CPR) is performed infrequently in Japan. We conducted this study to identify Japanese attitudes toward the performance of bystander CPR. METHODS: participants were asked about their willingness to perform CPR with varying scenarios and CPR techniques (mouth to-mouth ventilation plus chest compression (MMV plus CC) versus chest compression alone (CC)). RESULTS: a total of 1302/1355 individuals completed the questionnaire, including high school students, teachers, emergency medical technicians, medical nurses, and medical students. About 2% of high school students, 3% of teachers, 26% of emergency medical technicians, 3% of medical nurses and 16% of medical students claimed they would 'definitely' perform MMV plus CC on a stranger. However, 21-72% claimed they would prefer the alternative of performing CC alone. Respondents claimed their unwillingness to perform MMV is not due to the fear of contracting a communicable disease, but the lack of confidence in their ability to perform CPR properly. CONCLUSION: in all categories of respondents, willingness to perform MMV plus CC for a stranger was disappointingly low. Better training in MMV together with teaching awareness that CC alone can be given should be instituted to maximize the number of potential providers of CPR in the community, even in communities where the incidence of HIV is very low. PMID- 10825620 TI - Checking the carotid pulse: diagnostic accuracy in students of the healthcare professions. AB - This study evaluated the competence of students of the healthcare professions to locate the carotid pulse using a computerised manikin, within 10 s. A sample of 105 students from physiotherapy, radiography, midwifery and nursing participated in measuring diagnostic accuracy in a single attempt at pulse check using a computerised manikin, timed to an accuracy of +/-1 s. All had received basic life support instruction, and one group had advanced life support skills. The mode and median diagnostic delays were calculated for each group. Comparisons of mean rank values for the groups were determined, and comparisons of previous training and accuracy in diagnosis were calculated. Forty (38%) students were able to give an accurate diagnosis within 10 s. The results identified significant differences between the performance of the groups (chi(2) 16.74, P<0.01), with the advanced life support course students demonstrating most competence. Previous training did not affect performance in the skill (chi(2) 0.29, P=0.58). Carotid pulse check skills should be emphasised and tested as part of cardiopulmonary resuscitation instruction. PMID- 10825621 TI - Reliability of ECG monitoring with a gel pad/paddle combination after defibrillation. AB - Recent warnings have highlighted the possibility of unreliable monitoring through gel pads and paddles when using manual defibrillators. Occasionally, this causes an apparent asystole to be displayed when the true rhythm is ventricular fibrillation. We investigated this phenomenon in the laboratory using defibrillator-testing devices with two different impedances and with two defibrillators (Physio-Control LIFEPAK 9 and Hewlett Packard XL). After delivery of a 200 J shock, the time taken for the original ECG test signal to return to the defibrillator monitor was measured. Measurements were made after each of a series of ten shocks delivered with each defibrillator and gel pad/ testing device combination. Additional measurements were made using self-adhesive combination defibrillator electrodes. When using a low-impedance testing device with gel pads, on all occasions the initial rhythm reappeared immediately. When using the high-impedance testing device, the post shock rhythm was initially displayed as 'no signal' (Hewlett Packard XL) or 'asystole' (Physio-Control LIFEPAK 9). Over the series of ten shocks, the time to return of the original signal ranged between 24 and 154 s with the Hewlett Packard and 17-61 s with the Physio-Control LIFEPAK 9. The time for return of the signal increased with successive shocks. When using Fast-Patch electrodes, the original test signal always returned immediately. We conclude that after the delivery of a shock, monitoring through gel pads may result in the display of spurious asystole being displayed. This is more likely to occur in the presence of high chest impedance and with an increasing number of shocks delivered through the same gel pads. If defibrillator paddles and gel pads have been used for 'quick-look' monitoring, and 'asystole' is displayed after delivery of a shock, the rhythm should be confirmed immediately with monitoring leads. PMID- 10825622 TI - Responses to reversible anoxia of intracellular free and bound Ca(2+) in rat cortical slices. AB - Severe anoxia induces destabilisation of intracellular calcium homeostasis in neurones. The mechanism of this effect, and particularly the interrelationship between changes in intracellular concentration of free Ca(2+) ions and the content of the intracellular Ca(2+) stores, during and after anoxia, is not clear. We used a superfusion system of rat olfactory cortical slices for the fluorimetric estimation of changes in the intracellular concentration of free Ca(2+) ions and in the level of bound Ca(2+), utilising the fluorescent indicators Fura-2 and chlortetracycline, respectively. It was found that 10-min normoglycaemic anoxia results in simultaneous decrease in bound and increase in free Ca(2+) levels, whereas during 60-min reoxygenation, we detected an increase in both indices. The NMDA receptor antagonists MK-801 and APV attenuated changes in free Ca(2+) level during anoxia and reoxygenation and intensified anoxia evoked decrease in bound Ca(2+) content, whereas a late post-anoxic increase in bound Ca(2+) was abolished. These data suggest that the influx of extracellular Ca(2+) to neurones via NMDA receptors, plays a critical role in the rise of intracellular free Ca(2+) concentration during and after anoxia. Biphasic changes in bound Ca(2+) content during anoxia and reoxygenation may reflect an anoxia induced release of Ca(2+) from intracellular stores, followed later by a neuronal calcium overload and refilling of intracellular Ca(2+) binding sites. PMID- 10825623 TI - Successful recovery after ventricular fibrillation in a patient with Kawasaki disease. AB - Kawasaki disease (KD) is an uncommon cause of sudden death in young adults in Europe. Angiographically, the disease is characterized by coronary artery aneurysms which can be fully obstructed by acute thrombosis or by progression of the disease. If diagnosis of KD is made, immediate investigation should be made to determine whether ischemia is occurring and if so, to establish optimal time for revascularisation or cardiac transplantation. We describe an 18-year-old Caucasian male who was not previously known to have KD and who suffered from an acute myocardial infarction complicated by ventricular fibrillation, caused by acute thrombosis of a coronary artery aneurysm. PMID- 10825624 TI - Use of an inspiratory impedance threshold valve during cardiopulmonary resuscitation: a progress report. AB - Building upon studies on the mechanism of active compression-decompression (ACD) cardiopulmonary resuscitation, a new inspiratory impedance threshold valve has been developed to enhance the return of blood to the thorax during the decompression phase of CPR. Use of this device results in a greater negative intrathoracic pressure during chest wall decompression. This leads to improved vital organ perfusion during both standard and ACD CPR. Animal and human studies suggest that this simple device increases cardiopulmonary circulation by harnessing more efficiently the kinetic energy of the outward movement of the chest wall during standard CPR or active chest wall decompression. When used in conjunction with ACD CPR during clinical evaluation, addition of the impedance valve resulted in sustained systolic pressures of greater than 100 mmHg and diastolic pressures of greater than 55 mmHg. The new valve may be beneficial in patients in asystole or shock refractory ventricular fibrillation, when enhanced return of blood flow to the chest is needed to 'prime the pump'. The potential long-term benefits of this new valve remain under investigation. PMID- 10825625 TI - 'RESUS' for resus. PMID- 10825627 TI - Millennium "Issue of EJOGR" PMID- 10825628 TI - From anemia to spina bifida - the story of folic acid. A tribute to Professor Richard Smithells. PMID- 10825629 TI - Reinier De Graaf and his contribution to reproductive biology. PMID- 10825630 TI - Elizabeth M. Ramsey and the evolution of ideas of uteroplacental blood flow and placental gas exchange. AB - During the past century a number of investigators have contributed to an understanding of the regulation of uteroplacental blood flow and its influence on placental respiratory gas exchange and fetal oxygenation. Among these, Elizabeth M. Ramsey is noteworthy for her contributions to embryology and placental development. In addition, with several colleagues she performed pioneering cineangiographic studies on the patterns of blood flow in the primate placenta. PMID- 10825631 TI - The Pap test: George N. Papanicolaou (1883-1962). A screening test for the prevention of cancer of uterine cervix. PMID- 10825632 TI - William John Little and cerebral palsy. A reappraisal. PMID- 10825633 TI - Intrauterine contraception: from silver ring to intrauterine contraceptive implant. AB - Grafenberg introduced his silver ring in 1928 and by his classic paper, published in 1931, provided intrauterine contraception with a scientific basis. For largely non-medical reasons, the Grafenberg ring vanished from the scene a few years later, and it took almost 30 years before the method was rehabilitated as a result of a change in mental attitude toward birth control, the awareness of world population explosion, and the introduction of improved intrauterine devices (IUDs). The United States became the epicenter of IUD research in the 1960s but, due to adverse publicity and the threat of litigation, the pendulum swung and European investigators took over the job of improving the IUD. Proceeding from Dr Grafenberg's classic paper, the major steps in the evolution of the intrauterine devices are retraced with focus on the scientists who were innovative in the field. PMID- 10825634 TI - Semmelweis: the combat against puerperal fever. AB - Groups A and B streptococci are of great significance in the history of obstetrics. Group A streptococci were a great threat to the puerperium, especially in the 19th century, when homebirth was replaced by institutional birth in lying-in hospitals. The history of the rise and fall of puerperal fever is indeed a tragedy. Some people, like Semmelweis, who brought new and important evidence based findings were not believed by their fellow obstetricians, an attitude that spoiled thousands of innocent lives. Even today group A streptococci, though seldom, may be the cause of puerperal sepsis. Group B streptococci are widespread and may cause sepsis and important lifelong morbidity or mortality of the newborn. Obstetricians today try to establish cost-effective prophylactic measures during labor to prevent these neonatal infections. PMID- 10825635 TI - Endometriosis. PMID- 10825636 TI - Prenatal diagnosis of genetic diseases. PMID- 10825637 TI - Ethics and assisted reproduction. AB - In vitro fertilization and assisted reproductive technology have made great progress during the last 20 years. Genetic material donation, human embryo cryopreservation, selective embryo reduction, preimplantation genetic diagnosis and surrogacy are currently practiced in many countries. On the other hand, embryo research is practiced only in a small number of nations, whereas human cloning has thus far been universally condemned. The rapid evolution and progress of all these techniques of assisted reproduction has revealed certain ethical issues that have to be addressed. PMID- 10825638 TI - Leon C. Chesley and hypertension in pregnant women. AB - Leon C. Chesley's first paper opened with the title 'Pregnancy in the patient with hypertensive disease'(1). Leon C. Chesley PhD, John E. Annitto, MD, MSc (Med), Jersey City, NJ, USA (from the Margaret Hague Maternity Hospital) [Am J Obstet Gynecol 1947;53:372-381]. We quote some lines from this important paper and pioneering work: "We have thought it worth while to survey our experience with pregnancy in women with hypertensive disease. There are relatively few such studies based upon any considerable series, and in most studies extant there has been a selective factor in that therapeutic abortion has been done in the more severely hypertensive patients. It has not been our policy to abort such women, and our large material therefore offers an almost unique opportunity for the study of the natural history of pregnancy in hypertensive women. From the opening of the hospital in October 1931 through to 1944 there were a total of 218 patients in whom recorded blood pressures established the diagnosis of 'hypertensive toxemia,' as defined by the American Committee on Maternal Welfare. A detailed analysis has been made of the 301 pregnancies in which these patients have been seen. The gross fetal loss: in prior pregnancies, 35%; in first hypertensive pregnancy, 38%; in subsequent pregnancies, 40%. Of 47 sisters of these hypertensive patients, who delivered here, 45% had at least one toxic pregnancy. Nearly 40% of the hypertensive patients showed drops in the blood pressure in midpregnancy. Proteinuria of some degree occurred in half of the pregnancies. Renal function was normal in 93% of the pregnancies. Premature separation of the placenta occurred in 5.6% of the pregnancies. Fetal loss increased with: higher initial blood pressure, second trimester rises in blood pressure, higher pressures near delivery, decreased renal function, proteinuria, and superimposed toxemia. There were six immediate maternal mortalities (2.0%) and seven late puerperal deaths. Thus the mortality was 20 times that of the whole hospital experience." From here we will start our "hypertensive journey". PMID- 10825639 TI - Ultrasound scanning - Prof. Ian Donald (1910-1987). PMID- 10825640 TI - Ca2+ signaling in gerbil CA3 hippocampal neurons following transient in vivo ischemia. AB - Using the gerbil model of post-ischemic neuron death in the hippocampal CA1 region, it was recently shown that there is a strong down-regulation of voltage gated Ca2+ influx in neurons examined at 2 days after the ischemic insult (Connor, J.A., Razani-Boroujerdi, S., Greenwood, A.C., Cormier, R.J., Petrozzino, J.J. and Lin, R.C., Reduced voltage-dependent Ca2+ signaling in CA1 neurons after brief ischemia in gerbils, J. Neurophysiol., 81 (1999) 299-306). The aim of the present study was to determine whether a similar change occurs in pyramidal neurons of the CA3 region that are relatively resistant to transient ischemia. In vitro intracellular recordings and fluorometric Ca2+ measurements were made from CA3 neurons in coronal slices prepared from controls and 1 or 2 days following in vivo ischemia. In slices from control and post-ischemic animals, the electrophysiological properties of CA3 neurons were consistent with significant voltage-gated Ca2+ influx, leading to spike frequency adaptation. Quantitative results indicated no significant difference in Ca2+ transients evoked by action potential trains. This Ca2+ signaling was compared with responses in CA1 neurons from the same preparations, which showed substantially diminished Ca2+ influx at 2 days post-ischemia. These findings suggest that diminished Ca2+-signaling is not a general feature of pyramidal neurons following ischemia, but is characteristic of neurons destined to die. PMID- 10825641 TI - Prepulse inhibition of the blink reflex by laser stimuli in normal humans. AB - The subcortical integrative effects of laser-induced activation of pain ascending tracts were examined in 11 healthy volunteers, aged 22-52 years. Subjects underwent either CO2 laser stimulation at the dorsum of the hand, electrical stimulation of digital nerves at the 3rd finger, or mechanical taps to the first dorsal interosseous space, preceding a blink reflex elicited by a supraorbital nerve electrical stimulus. The percentage inhibition induced in the R2 response of the blink reflex was similar for the three different stimulus modalities, but occurred at a different time interval. Compared to control trials, the R2 response of the test trials was a mean of 23.1% at the interval of 250 ms with laser stimuli, 17.4% at the interval of 100 ms with electrical stimuli to the 3rd finger, and 20.6% at the interval of 90 ms with a mechanical tap to the 1st interosseous space. Activation of pain receptors induces prepulse inhibition of the blink reflex at a delay corresponding to a slowly conducting pathway. The percentage inhibition is similar to that observed with other somatosensory inputs. PMID- 10825642 TI - Rapid recovery (20 ms) of human 600 Hz electroencephalographic wavelets after double stimulation of sensory nerves. AB - Non-invasive scalp-recordings of human somatosensory evoked potentials (SEP) contain high-frequency (600 Hz) wavelet bursts, presumably generated by synchronized thalamocortical and/or intracortical population spikes. Here, double pulse stimulation (interval 20 ms) in 12 healthy subjects revealed significantly different burst recovery for mixed vs. sensory-only nerves. For median nerves the second burst response was decreased (11/11 subjects), possibly due to interfering reafferent (e.g. muscle spindle) input. In contrast, for sensory-only superficial radial nerves (containing less fibers than median nerves), weak bursts were detected in 6/11 subjects and were found fully recovered in 4/6 subjects. This potential for rapid burst recovery at 20 ms intervals renders contributions from neurons emitting bursts based on slowly recovering low-threshold calcium spikes unlikely and favors the generation of macroscopic SEP bursts by specialized cell populations, e.g. inhibitory interneurons and/or chattering cells the latter of which are capable to discharge rapidly repeating (50 Hz) high-frequency (600 Hz) bursts of fast sodium spikes. PMID- 10825643 TI - Vitamin E deficiency has different effects on brain and liver phospholipid hydroperoxide glutathione peroxidase activities in the rat. AB - The effect of vitamin E deficiency on glutathione peroxidase activity (GPX) and on the activity of a selenoenzyme (phospholipid hydroperoxide glutathione peroxidase (PHGPX) was measured in rat brain and liver. In brain, the activity of both enzymes was in the same range in homogenate and in microsomes. In contrast, in liver homogenate, PHGPX activity was approximately 20 times lower than that of GPX. Very interestingly, PHGPX activity was significantly decreased in brain microsomes by vitamin E deficiency, but slightly significantly increased in liver microsomes. In contrast, GPX activity was not affected in brain by vitamin E deficiency, but was significantly lower in liver homogenate and microsomes. Thus, PHGPX activity is partially controlled by vitamin E in membranes, and PHGPX is probably an enzyme different from GPX. PMID- 10825644 TI - Role of frontal cortex in memory for duration: an event-related potential study in humans. AB - Event-related potentials were recorded in order to determine how brain activity is lateralized during the encoding and the recognition of visual durations (700 and 2500 ms ranges). It is assumed in the Hemispheric Encoding Retrieval Asymmetry model that the encoding of words, faces and odours involves left frontal areas whereas their recognition involves right frontal areas. The present results indicate that, for temporal information, the hemispheric bias is different: a negativity developed over right frontal electrodes for both encoding and recognition, and for both duration ranges. Thus, the involvement of right frontal areas appears critical for time perception. Conversely to what was expected, contingent negative variation during recognition was large over both left and right frontal electrodes. These results suggest that the involvement of both hemispheres is necessary for recognition of temporal information. PMID- 10825645 TI - Spinal CSF from rats with painful peripheral neuropathy evokes catecholamine release from chromaffin cells in vitro. AB - The environment presented by host tissue may influence cellular transplants in the CNS depending on injury or disease. Here we examined whether chronic pain alters cerebrospinal fluid (CSF), thereby enhancing the analgesic effect of transplanted adrenal cells. CSF samples were taken intracisternally from rats with neuropathic pain induced by chronic constriction injury of the sciatic nerve. The samples were applied to cultured bovine chromaffin-cell clusters while catecholamine release was measured by fast cyclic voltammetry. This caused marked and sustained elevations in catecholamine levels, compared to CSF from sham operated controls, which were reversible by the nicotinic antagonist mecamylamine. These results suggest that chronic neuropathic pain produces increased CSF levels of secretogogues for chromaffin cells, and illustrates the importance of host microenvironmental factors in determining graft function. PMID- 10825646 TI - Appearance of tau-2 immunoreactivity in glial cells in human brain with cerebral infarction. AB - Appearance of tau epitopes in ischemic foci was investigated immunohistochemically on a series of autopsied human brains using a panel of anti tau antibodies. While neurons were immunopositive for Alz-50, microglia and oligodendroglia around ischemic foci abundantly contained tau-2 immunoreactivity. Some astrocytes contained tau-2 immunoreactive granules in their cytoplasm. This difference suggests that neurons and glia react differently to an ischemic insult by exhibiting different tau epitopes. Immunohistochemical visualization tau-2 epitope represents its conformational change, as was reported with neurofibrillary tangles of Alzheimer type. Lack of argyrophilia in any of these tau-immunoreactive cells distinguishes them from tau-immunoreactive structures seen in various neurodegenerative disorders. PMID- 10825647 TI - Co-existence of calcium-binding proteins in neurons of the medullary dorsal horn of the rat. AB - Double immunofluorescence histochemistry for calcium-binding proteins was performed in the caudal subnucleus of the rat spinal trigeminal nucleus; for calbindin D28k (CB) and calretinin (CR), for CB and parvalbumin (PV), and for CB and CR. CB-immunoreactive (-ir) neurons were seen 1.7 times more frequently than CR-ir neurons and 5.5 times more frequently than PV-ir neurons. About 70-90% of these neurons were distributed in substantia gelatinosa. Co-existence of CB and CR was indicated in 2.3% of CB-ir and 3.9% of CR-ir neurons. Co-existence of CB and PV was indicated in 1.0% of CB-ir and 5.5% of PV-ir neurons. Co-existence of CR and PV was indicated in 1.4% of CR-ir and 5.1% of PV-ir neurons. In these doubly immunostained neurons, 59.5-69.5% were observed in substantia gelatinosa, 5.9-17.8% in the marginal zone, and 12.7-31.0% in the magnocellular part. PMID- 10825648 TI - Effects of estrogen on the expression of connexin32 and connexin43 mRNAs in the suprachiasmatic nucleus of female rats. AB - The suprachiasmatic nucleus of the hypothalamus (SCN) is the site of the endogenous biological clock that regulates circadian rhythms in mammals. It has been shown that the gap junction couples neurons in the rat SCN. To determine the effects of estrogen on gap junction communication in the SCN, we examined the effects of estrogen on the expression of connexin32 and connexin43 mRNAs in the SCN and cerebral cortex (CX) of female rats. Ovarectomized adult rats were injected with 20 microg of 17beta-estradiol and killed 24 h after the treatment. Northern blot revealed that the expression of connexin32 mRNA was significantly increased in the SCN but not in the CX. On the other hand, estrogen increased the connexin43 mRNA level in the CX but not in the SCN. These results suggest that the gap junction with connexin32 is specifically regulated by estrogen in the SCN of female rats. PMID- 10825649 TI - Gamma-aminobutyric acid enhancement of halothane binding in rat cerebellum. AB - Quantitative autoradiography of 14C-halothane direct photolabeled rat cerebellum sections was performed in the presence of increasing concentrations of gamma aminobutyric acid (GABA) or glutamate to test the hypothesis that a coupled binding equilibrium between the anesthetic and neurotransmitter exists. The results show that halothane binding was enhanced in the presence of GABA by approximately 50% in the molecular layer and to a lesser extent in the granular layer, with no change in the myelin layer. Glutamate, however, did not enhance halothane binding in any layer. These data confirm the presence of coupling, and thus suggest a direct interaction of halothane with a GABA binding protein. PMID- 10825650 TI - Brain dynamics in theta and alpha frequency bands and working memory performance in humans. AB - In this study non-linear and linear global electroencephalogram (EEG) changes during a visual working memory task were studied using a separate analysis of theta, lower alpha and upper alpha band filtered data. EEGs were recorded in 21 healthy subjects (62.5 year; SD 2.1; 12 females, nine males) during an eyes closed no-task condition and a working memory condition. Coarse-grained dimension was estimated for both conditions from spatially embedded EEG data filtered in the theta band and both alpha bands. Linear measures of coupling and mean amplitude were also computed. During the working memory condition lower alpha band dimension increased. Linear analysis showed alpha1 band desynchronization. Female subjects had a higher dimension in the theta band as well as more desynchronization in the theta and alpha1 band. Working memory capacity correlated with a lower theta band dimension during the no-task condition in female subjects. The increase in alpha1 band complexity can be interpreted as increased desynchronization corresponding with attentional processes. Higher complexity/desynchronization in females seems to be a more structural phenomenon and may be more intimately related to task performance. PMID- 10825651 TI - Dopamine inhibits melatonin release in the mammalian retina: in vitro evidence. AB - A circadian oscillator located within the retina controls melatonin synthesis in the retina of mammals. In non-mammalian vertebrates retinal melatonin and dopamine appear to act as mutually inhibitory paracrine signals for night and day, respectively; while in mammals this mutually inhibitory capability has now been fully demonstrated. In this study using a flow-through culture apparatus we investigated melatonin release from cultured retinas of golden hamster (Mesocricetus auratus) in the presence of dopamine or dopaminergic agonists and antagonists. Neural retinas were cultured with medium 199 for 24 h in a flow through apparatus at the temperature 33 degrees C. During the subjective night the culturing medium was supplemented with dopamine, dopamine receptor antagonists or agonists. At the concentration of 0.1 microM dopamine did not inhibit melatonin release, while at higher dopamine concentration (1 to 1000 microM) melatonin release was inhibited in a dose-dependent manner. These effects appeared to be mediated by a D2/D4 receptor, because D2 and D4 receptor agonists (100 microM), but not D1/D5 receptor agonists (100 microM), inhibited melatonin release. Furthermore, D2/D4 selective receptor antagonists (100 microM) in conjunction with 100 microM dopamine blocked melatonin suppression, whereas a D1/D5 selective receptors antagonist was completely ineffective. Taken together, these results directly demonstrate for the first time that in the retina of mammals dopamine may suppress melatonin, and that suppression is mediated by D2/D4 dopaminergic receptors. PMID- 10825652 TI - The role of tachykinin NK-1 receptors in the area postrema of ferrets in emesis. AB - The role of tachykinin NK-1 receptors in the area postrema (AP) in emesis was examined in ferrets. Strong c-fos-like immunoreactivity was observed in the AP and nucleus tractus solitalius (NTS) in cisplatin (10 mg/kg, i.p.)-treated animals, but not in control animals. The number of the central emetogen morphine induced vomits and retches was remarkably reduced (95%) and that of the peripheral emetogen copper sulphate-induced vomits was significantly (54%) reduced by AP lesion. Pretreatment with the tachykinin NK-1 receptor antagonists HSP-117 (1.0 microg) and CP-99,994 (7.5 microg) into the AP decreased the numbers of vomits and retches induced by morphine and copper sulphate. These results suggest that NK-1 receptors in the AP are involved in the mechanism of emesis induced by morphine and copper sulphate. PMID- 10825653 TI - Upper cervical afferents to the motor trigeminal nucleus and the subnucleus oralis of the spinal trigeminal nucleus in the rat: an anterograde and retrograde tracing study. AB - Upper cervical afferents to the motor trigeminal nucleus (Vmo) and the subnucleus oralis (Vo) neurons projecting contralaterally to the cervical cord were demonstrated in the rat. Axon-terminals were labeled with biotinylated dextran and neurons with cholera toxin subunit B. Axons from the C2 and C3 segments terminated ipsilaterally on the somata and proximal dendrites of Vmo neurons. In the Vo, terminals of axons from the C2 and C3 segments were densely distributed on the somata, and proximal to distal dendrites of neurons projecting contralaterally to the cervical cord. The ipsilateral cervical input to the Vmo would modulate the activity of motoneurons of masticatory muscles while that to the Vo neurons subserves the feedback control of the trigemino-spinal reflex. PMID- 10825654 TI - Inhibition of monkey brain semicarbazide-sensitive amine oxidase by various antidepressants. AB - We examined whether the antidepressant drugs, such as the dicyclic drug zimeldine, the tricyclic drug imipramine, tetracyclic drug maprotiline, and the non-cyclic drug nomifensine, inhibit in vitro semicarbazide-sensitive amine oxidase (SSAO) activity in monkey brain. The deamination of 1 microM benzylamine was not inhibited at high concentrations of clorgyline or deprenyl, while it was highly sensitive for semicarbazide. When corresponding experiments were performed with 100 microM benzylamine, the opposite results were obtained. The most potent of inhibition of SSAO was observed by imipramine, followed by maprotiline, zimeldine and nomifensine. Inhibition of SSAO was not enhanced by varying the time of preincubation of the enzyme and various antidepressant drugs, indicating direct action on and reversible inhibition of SSAO. We found the tricyclic antidepressant drug to be the most selective inhibitors of SSAO activity in monkey brain, as compared with other type of antidepressant drugs. PMID- 10825655 TI - Inactivation of alpha7 ACh receptors and activation of non-alpha7 ACh receptors both contribute to long term potentiation induction in the hippocampal CA1 region. AB - Acute and chronic nicotine exposure differentially facilitate the induction of long-term potentiation (LTP), a synaptic model of learning and memory, in the hippocampal CA1 region. The mechanisms underlying these effects of nicotine, however, are unknown. In the present study, both nicotinic acetylcholine receptor (nAChR) agonists and an alpha7 nAChR antagonist facilitated the induction LTP in the hippocampal CA1 region of naive rat. Furthermore, chronic nicotine treatment lowered the threshold for induction of LTP, and acute application of nicotinic agonists, but not an alpha7 antagonist, further facilitated LTP induction in the chronic-nicotine-treated hippocampus. These results suggest not only that both activation of non-alpha7 nAChRs and inactivation of alpha7 nAChRs contribute to LTP induction, but also that chronic-nicotine-mediated facilitation of LTP induction is due to chronic-nicotine-induced desensitization of alpha7 nAChRs. PMID- 10825656 TI - Daily changes of cytochrome oxidase activity within the suprachiasmatic nucleus of the Syrian hamster. AB - Cytochrome oxidase (CO) activity was studied over a 24-h period in the Syrian hamster suprachiasmatic nucleus (SCN) (site of the biological clock), anterior hypothalamic area (AHA), and motor cortex. The SCN CO activity was highest at the middle of the day (Zeitgeber time (ZT) 05), decreased at the end of the light period (ZT 10) and continued at a low level during the night (ZT 13, 16, 21.5 and 24). AHA and motor cortex showed a similar profile of CO activity and no changes of CO activity were found in animals maintained under darkness (DD). We propose that photic input plays a role in the SCN neuronal activity that modulates metabolic activity on this area. PMID- 10825657 TI - First report of polymorphisms in the prion-like protein gene (PRND): implications for human prion diseases. AB - The aim of this study was to investigate the possible involvement of genetic variation in the prion-like protein gene (PRND), which encodes the doppel protein (Dpl), in the aetiology of human prion diseases. Patients with sporadic, infectious or genetic forms of human prion diseases and controls were systematically screened, using the single-strand conformational polymorphism method, for genetic variants of the PRND gene. Four polymorphisms in PRND (three structural changes, T26M, P56L and T174M and a silent polymorphism, T(174)T) were detected. No strong association was found between any of these polymorphisms and human prion diseases but certain PRND alleles may be useful markers for tracing the chromosomal ancestry of PRNP mutations. Although genetic variation in PRND does not seem to play a major role in the pathogenesis of prion diseases, this first report of PRND polymorphisms may open up new possibilities for investigating the involvement of such polymorphisms in other human diseases. PMID- 10825658 TI - Calpain-mediated degradation of PSD-95 in developing and adult rat brain. AB - PSD-95 is a major postsynaptic density protein that is degraded as a result of synaptic activity. We used four different methods to test the hypothesis that calpain is involved in PSD-95 turnover. Treatment of synaptic membranes with purified calpain resulted in a decrease in immunoreactivity of the native 95 kDa protein and the appearance of two smaller molecular weight species, migrating at 50 and 36 kDa, respectively. Calcium treatment of frozen-thawed brain sections produced an identical digestion pattern, an effect blocked by calpain inhibitors. N-methyl-D-aspartate treatment of organotypic hippocampal cultures produced truncation of PSD-95 and accumulation of the 36 kDa species. Finally, calpain generated degradation products of PSD95 were prominent in neonatal hippocampus, and disappeared with postnatal development. Our data suggest that PSD-95 is a substrate for calpain, and that calpain-mediated truncation contributes to PSD-95 turnover. PMID- 10825659 TI - A dimeric creatine kinase from a sponge: implications in terms of phosphagen kinase evolution. AB - This study demonstrates conclusively that tissues of the sponge Tethya aurantia contain significant creatine kinase (CK) activity. This CK was purified and analyzed with respect to a number of physico-chemical properties. Size exclusion chromatography and denaturing gel electrophoresis analyses showed that this enzyme is dimeric. The sequences of several Lys-C endoproteinase peptides from Tethya CK are consistent with this enzyme being a member of the phosphagen kinase family and a true CK. CK in higher organisms exists in a variety of quaternary structure forms--dimer, octamer and large monomer consisting of a three contiguous CK domains. The present results indicate that CK evolved very early in metazoan evolution and that the dimeric structure preceded other subunit association forms. PMID- 10825660 TI - Partial characterization of alpha-amylase in the salivary glands of Lygus hesperus and L. lineolaris. AB - The alpha-amylases in the salivary glands of Lygus hesperus Knight and L. lineolaris (Palisot de Beauvois) were isolated and purified by ion exchange chromatography, and by isoelectric focusing, respectively. The alpha-amylase from L. hesperus had an isoelectric point (pI) of 6.25, and a pH optimum of 6.5. The specific activity of alpha-amylases in the salivary glands of L. hesperus was 1.2 U/mg/ml. The alpha-amylase from L. lineolaris had a pI of 6.54, and a pH optimum of 6.5. The specific activity of alpha-amylase from L. lineolaris was 1.7 U/mg/ml. The activity of alpha-amylase in both species was significantly inhibited by alpha-amylase inhibitor from wheat and also by EDTA and SDS. Sodium chloride enhanced alpha-amylase activity for both species. The enzyme characteristics and relative activities are discussed in the context of differences phytophagous versus zoophagous habits in these two congeneric species. PMID- 10825661 TI - cDNA cloning and expression of two Ki-ras genes in the flounder, Platichthys flesus, and analysis of hepatic neoplasms. AB - The screening of a flounder cDNA library with a partial sequence of ras gene from flounder (exons 1 and 2) allowed the isolation of two complete cDNA sequences (ras1 and ras2) highly homologous to human Ki-rasb genes. ras1 and ras2 sequences have an homology of 77.3% indicating that they represent two distinct genes, which differ particularly in their 3 regions. ras1 and ras2 intron 1 sequencing revealed an homology of only 50%, confirming that they represent two different genes. Both genes encode for a 188 amino-acid protein, a size characteristic of Ki-rasb proteins. ras1 protein has the stronger homology to the human Ki-rasb protein (99% identity) and ras2 presents a 85.5% of homology. Two transcripts of respectively 2 and 2.8 kb were identified by northern blots with either ras1 or 2 probes. Preneoplastic and neoplastic livers collected from 14 flounder did not present any mutation on the ras2 gene. PMID- 10825662 TI - Purification and characterization of a dipeptidyl carboxypeptidase from the polychaete Neanthes virens resembling angiotensin I converting enzyme. AB - Dipeptidyl carboxypeptidase (DCP) is well known as a mammalian angiotensin I converting enzyme (ACE) which plays an important role in blood pressure homeostasis. DCP was purified from the whole body of a polychaete, Neanthes virens. The purified enzyme was homogeneous by SDS-PAGE, with a molecular mass of 71 kDa by SDS-PAGE and 69 kDa by gel filtration, indicating that it is monomeric. The isoelectric point was 4.5 and optimum pH for the activity was 8.0. It showed a specific activity of 466.8 U/mg, which is the highest of known DCPs. The enzyme hydrolyzed angiotensin I to angiotensin II and sequentially released Phe-Arg and Ser-Pro from the C-terminus bradykinin, but does not cleave imido-bonds. Activity was completely inhibited by 1 mM EDTA and 5 mM o-phenanthroline, but it was not affected by serine and aspartic protease inhibitors. The original activity of EDTA-inactivated DCP was restored by addition of cobalt, manganese or low concentrations of zinc. The Km and Vmax values of the enzyme for Bz-Gly-His-Leu were 0.56 mM and 600 mumol/min per mg, respectively. The Ki values for specific mammalian ACE inhibitors, such as captopril and lisinopril, were 1.38 and 2.07 nM, respectively. In conclusion, we have shown the existence of a DCP from the polychaete, N. virens, with similar properties to those of mammalian ACE. PMID- 10825663 TI - Lipid composition of blood platelets and erythrocytes of southern elephant seal (Mirounga leonina) and antarctic fur seal (Arctocephalus gazella). AB - Erythrocyte and blood platelet phospholipid compositions were studied in three elephant seals and two fur seals, two species of marine mammals living in the Subantarctic region feeding on preys rich in (n-3) polyunsaturated fatty acids. Results were compared with those reported for related species and humans. In erythrocytes, the phospholipid (PL) and cholesterol (CHOL) contents were lower in pinnipeds than in humans. Phosphatidylcholine (PC) levels were higher in elephant seals than in fur seals, with a reverse trend for phosphatidylethanolamine (PE) and phosphatidylserine (PS). Both species had lower SM/PC ratios and PE plasmalogen concentrations than human. Erythrocytes were richer in (n-3) fatty acids (FA) in pinnipeds than in humans. In platelets, the PL content was lower and the CHOL content higher in elephant seals than in humans or in other phocid seal species studied to date. The SM/PC ratio was much higher than in other seal species or in man. In both species, the proportion of PE plasmalogens was higher in platelets than in erythrocytes. PL were more saturated in elephant seals than in fur seals. These results suggest that the erythrocytes and platelets of wild marine mammals may prove useful models to study the influence of dietary lipids on the structure and hemostatic function of these cells. PMID- 10825664 TI - Molecular characterization of the first non-mammalian p73 cDNA. AB - The p53 gene is believed to be mutated or deficient in over 50% of human tumours, and is therefore considered to be instrumental in the process of carcinogenesis. Recently in humans, homologues of p53 (such as p73 and p63) have been isolated. In our studies in fish, we have been isolating tumour suppressor genes with a view to their potential use to study genotoxins in the aquatic environment. In this paper, we report the characterisation of the first non-mammalian p73 cDNA, isolated from barbel (Barbus barbus), a freshwater cyprinid fish indigenous to UK rivers. The deduced barbel p73 amino acid sequence has a high homology with human p73 alpha: the proteins are 641 and 636 aa in length, respectively, and there is a 72% identity over the entire sequence length of the protein (over 90% in the putative DNA binding domain). The level of conservancy for p73 is considerably higher across class (from man to fish), than for p53 and it may therefore have particular value in studies on environmental mutagenesis. Northern analysis showed expression of three p73 mRNA transcripts/homologues. The patterns of p73 tissue expression in the barbel differed from the expression of p53 mRNA, suggesting specific functional roles for the two genes. PMID- 10825665 TI - Abalone collagens: immunological properties and seasonal changes of their mRNA levels. AB - The antisera were raised against pepsin-solubilized abalone collagen and its corresponding gelatin. The reactivity against abalone collagen was higher with the anti-collagen than anti-gelatin antiserum. The two antisera recognized all type I collagens from various vertebrates, whereas these had no reactivity against vertebrate type III and type V collagens. Furthermore, both antisera reacted with only alpha 2(I) chains from chicken, rat, and calf. The strong reactivity was observed against the two antisera in the case of invertebrate and protochordate collagens, especially for turban shell collagen. The seasonal changes of collagen mRNA levels were examined in relation to those of collagen content. Haliotis discus collagens (Hdcols) 1 alpha and 2 alpha coding for abalone collagen pro alpha-chains showed quite similar patterns. The highest mRNA levels in adductor and foot muscles for the two collagens were observed in December and January, in good agreement with the increase of collagen content. The mRNA levels decreased in July and August when collagen content decreased. These results indicate that collagen transcription levels are closely related to collagen contents. PMID- 10825666 TI - Discrimination of Sebastes viviparus, Sebastes marinus and Sebastes mentella from Faroe Islands by chemometry of the fatty acid profile in heart and gill tissues and in the skull oil. AB - The composition of fatty acids in the tissue of heart, gill and skull oil of the three redfish species, Sebastes viviparus, Sebastes marinus and Sebastes mentella was determined by a chemometric method. The method consists of methanolysis of samples of the tissues and of the oils, gas chromatography of the resulting fatty acid methyl esters and multivariate statistical treatment, by principal component analysis, of the analytical data. Although the differences in fatty acid composition among the three tissues were the dominating features of the data, the three species had significantly different fatty acid profiles within each tissue, even though variation among the individuals was considerable. The fatty acid profiles appear to be species specific. The mutual relationship between S. marinus and S. mentella is closer than the relationship between either of them and S. viviparus. PMID- 10825667 TI - Role of hepatic lipogenesis in the susceptibility to fatty liver in the goose (Anser anser). AB - In response to overfeeding, the Landes goose develops a fatty liver that is twice as large as that of the Poland goose, despite similar food intake. The role of hepatic lipogenesis in the genetic susceptibility to fatty liver was assessed in male overfed geese of the two breeds. For a similar hepatic protein content, total activities of malic enzyme, glucose-6-phosphate dehydrogenase, acetyl-Coa carboxylase and fatty acid synthase, and specific activity and mRNA level of malic enzyme were about two-fold higher in the Landes goose. In the Poland goose, the weight of the fatty liver was correlated positively with the specific activity of ME and the VLDL concentration, which was not the case in the Landes breed. These results show that: (1) hepatic lipogenesis remains very active until the end of the overfeeding period; (2) the pentose-phosphate pathway may function in birds, contrary to what is assumed usually; (3) the level of hepatic lipogenesis is a major factor in the susceptibility to hepatic steatosis in different breeds of geese; and (4) ME activity may be a limiting factor of lipid synthesis in the less susceptible Poland breed. PMID- 10825668 TI - NAD(P)(+)-glycohydrolase from human spleen: a multicatalytic enzyme. AB - NAD(P)(+)-glycohydrolase (NADase, EC 3.2.2.6) was partially purified from microsomal membranes of human spleen after solubilization with Triton X-100. In addition to NAD+ and NADP+, the enzyme catalyzed the hydrolysis of several NAD+ analogues and the pyridine base exchange reaction with conversion of NAD+ into 3 acetylpyridine adenine dinucleotide. The enzyme also catalyzed the synthesis of cyclic ADP-ribose (cADPR) from NAD+ and the hydrolysis of cADPR to adenosine diphosphoribose (ADPR). Therefore, this enzyme is a new member of multicatalytic NADases recently identified from mammals, involved in the regulation of intracellular cADPR concentration. Human spleen NADase showed a subunit molecular mass of 45 kDa, a pI of 4.9 and a Km value for NAD+ of 26 microM. High activation of ADPR cyclase activity was observed in the presence of Ag+ ions, corresponding to NADase inhibition. PMID- 10825669 TI - alpha-Amylase from developing embryos of the camel tick Hyalomma dromedarii. AB - alpha-Amylase activity in the camel tick Hyalomma dromedarii was followed throughout embryogenesis. During purification of alpha-amylase III to homogeneity, ion exchange chromatography lead to four separate forms (termed I, II, III and IV). alpha-Amylase III with the highest specific activity was pure after chromatography on Sephacryl S-300. The molecular mass of alpha-amylase III was 106 kDa for the native enzyme, composed of two subunits of 43 and 66 kDa, respectively. alpha-Amylase had a value of 10 mg starch/ml. Varying alpha-amylase activity was detected when supplied with various substrates. alpha-Amylase III had a temperature optimum at 40 degrees C with heat stability up to 50 degrees C, and a pH optimum of 7.0. The enzyme activity was activated by CaCl2, MgCl2 and NaNO3, but not activated by NaCl, p-CMB, N-ethylmaleimide and iodoacetamide. EDTA and beta-mercaptoethanol strongly inhibited activity. PMID- 10825670 TI - Enolase from Candida albicans--purification and characterization. AB - This paper describes isolation of electrophoretically homogenous enolase from Candida albicans. The purification involved: disintegration of C. albicans cells in a Braun's mill (67-100%) ammonium sulfate precipitation, chromatography on DEAE-Sephadex A-50 at pH 9.0 and chromatography on CM-Sephadex A-50 at pH 6.2. The procedure resulted in a 30-fold purification of the enzyme with a recovery rate of 6% and a specific activity 35 U mg-1. The subunit molecular weight was 46 kDa and the pH optimum of the enzyme was 6.8. Km and Vmax values for the 2PGA- >PEP reaction were determined (Km = 0.95 mM, Vmax = 4200 mumol min-1 mumol-1). In the absence of orthophosphate, inhibition by fluoride was competitive, which became noncompetitive in the presence of phosphate. It was confirmed that Mg2+ is the most potent activator (Km = 0.286 mM); Mn2+ gave less activity and Zn2+ less still. It was also demonstrated that the presence of two types of cations in the reaction mixture nullified the activatory effect of the stronger agent. Properties of the enzyme from C. albicans are compared with those of enolases from other sources. PMID- 10825671 TI - Chemical testing strategies for predicting health hazards to children. AB - The United States Environmental Protection Agency has proposed the development of a Children's Health Test Program under the Toxic Substances Control Act. The Environmental Protection Agency's proposal for the children's health test battery has 12 different assays including general toxicity, genotoxicity, carcinogenicity, neurotoxicity, and developmental and reproductive toxicity. The current Environmental Protection Agency testing proposal is an "all or nothing" test battery. An alternative and preferable approach would be to use a science based, tiered testing scheme. It is proposed that the Screening Information Dataset program, currently used by the Organization for Economic Co-operation and Development (OECD) for the Screening Information Dataset-High Production Volume test battery, or equivalent, be considered for the first step. Step 1 would include acute and repeat dose toxicity testing, developmental toxicity testing (first species OECD 414 or OECD 422), reproductive toxicity screening (OECD 415 or 422), and genetic toxicity testing. For this step, the rat would be the initial and only species tested unless the mouse was used for in vivo genetic toxicity. Step 2 of the proposed children's health test battery would include developmental testing (second species OECD 414) or special mode of action studies performed for those chemicals that proved to be developmental toxicants in Step 1. Those chemicals that tested positive as reproductive toxicants in Step 1 would be tested in a two-generation reproduction study (OECD 416) or a special mode of action study. Steps 1 and 2 provide information on whether oncogenicity or developmental neurotoxicity testing is useful. Step 3 would include chronic toxicity/oncogenicity testing for those chemicals that tested positive for genetic toxicity in Step 1, and positive for developmental concerns in Step 2. In this step, chemicals would also be tested for developmental neurotoxicity if they showed evidence of neuropathy, behavioral effects, or neurotoxic potential in earlier studies. This stepwise approach would conserve resources and answer scientific questions in a logical, orderly, timely, and cost-effective manner. PMID- 10825672 TI - Six high-priority organochlorine pesticides, either singly or in combination, are nonestrogenic in transfected HeLa cells. AB - There have been increasing concerns that environmental chemicals may adversely affect the health of humans and wildlife by acting as endocrine modulators. These concerns have been augmented by the realization that human exposure occurs not just to single chemical agents, but typically to mixtures of chemicals that could exhibit estrogenic activity qualitatively and/or quantitatively different from that of individual components. To address these concerns, we have evaluated the ability of six organochlorine pesticides (4, 4'-DDT, 4,4'-DDD, 4,4'-DDE, aldrin, dieldrin, or endrin, all classified high priority by ATSDR) to modulate transcriptional activation of an estrogen-responsive reporter gene in transfected HeLa cells. In these assays, HeLa cells cotransfected with an expression vector encoding estrogen receptor and an estrogen-responsive chloramphenicol acetyltransferase (CAT) reporter plasmid were exposed to these pesticides individually and in defined combinations. While estradiol consistently elicited 10- to 23-fold dose-dependent inductions in these assays, the six organochlorine pesticides showed no detectable dose-related response when tested individually. When tested in binary combinations, the pesticide mixtures showed no additional estrogenicity. Thus, the pesticides tested, singly or as mixtures, showed virtually no evidence of estrogenicity. PMID- 10825673 TI - Lactoferrin is an estrogen responsive protein in the uterus of mice and rats. AB - To identify lactoferrin (LF) and determine its estrogen-responsiveness in the rat uterus, immature Sprague-Dawley rats were untreated or subcutaneously injected with 17beta-estradiol (500 microg/kg) for 3 days and uterine tissues collected. Outbred immature CD-1 mice, treated with 17beta-estradiol, provided the positive control. By using a polyclonal antibody raised against mouse LF, minimal detectable protein was immunolocalized in uterine epithelial cells of untreated immature rats and mice. After estrogen treatment, LF was localized in all uterine epithelial cells of both species, although staining was more intense in mice than rats. In mice, LF was evenly distributed throughout the cytoplasm with intense staining in some cells, while in rats, it was seen mainly in the apical cytoplasm. For comparison to another well-known estrogen responsive protein in rats, complement C3 was immunolocalized within epithelial cells and it showed a different staining pattern than LF. Uterine tissue homogenates were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blots showed cross-reactivity with the mouse LF antibody. These findings indicate that LF is present in the rat uterus, and is induced by estrogens as reported in other species. Thus, LF is an important marker of estrogenic activity across species and will, therefore, have utility in screening for effects of environmental estrogenic compounds. PMID- 10825674 TI - Assessment of estrogenicity by using the delayed implanting rat model and examples. AB - Endocrine disrupting chemicals have recently drawn increased interest. The delayed implanting rat model is a method that can identify and quantify the estrogenic activity of a chemical. In rats hypophysectomized after breeding, the administration of progesterone delays embryo implantation, and exposure to one dose of an estrogenic substance initiates implantation. Although methoxychlor was ineffective at dosages below 400 mg/kg when given by injection, the administration of the chemical by gavage resulted in an increase in the percent of fertilized rats exhibiting implantation sites. These results were statistically significant at dosages of 50, 100, 200, and 300 mg methoxychlor/kg. When bisphenol A was administered, by subcutaneous injection, dosages of 50, 100, and 200 mg/kg induced implantation. Only the 400 mg/kg dose of 4-tert-octylphenol was effective. Doses of beta-sitosterol up to 30 mg/kg failed to initiate implantation. These data confirm previous evidence of the availability of this model for evaluating estrogenic activity and provide estimates of the estrogenic potencies of several environmentally important chemicals. PMID- 10825675 TI - Dose- and treatment duration-related effects of p-tert-octylphenol on female rats. AB - We conducted a study to investigate the relation between estrogenic effects of p tert-octylphenol (OP) and serum concentration as well as treatment duration. Adult ovariectomized rats were given daily subcutaneous injections of OP for 2 and 14 days. 17beta-Estradiol was also administrated at a dose of 5 microg/kg. OP was detected in serum at doses of 25 mg/kg and above for 2 days and with multiple doses (14 days) of 12.5 mg/kg. Uterine weights and luminal epithelial height were increased although the effects were weak compared to 17beta-estradiol. Estrous conversion of the vaginal smear was detected only in 14-day-treated animals. OP treatment for 2 days caused a dose-related increase in proliferation of uterine luminal, glandular, and stromal cells and vaginal epithelial cells. From the 14 day experiment, the minimum estrogenic dose level of OP was concluded to be 25 mg/kg/day and the serum level at the dose was determined to be about 80 ng/ml. These findings demonstrated that OP exerts estrogenic activity in the female reproductive tract of ovariectomized rats only at high levels of exposure and that the effects are fundamentally related to serum OP levels. PMID- 10825676 TI - Effect of mancozeb on ovarian compensatory hypertrophy and biochemical constituents in hemicastrated albino rat. AB - Mancozeb a carbamate fungicide was administered orally at doses of 500, 600, 700, and 800 mg/kg/day to normal hemicastrated virgin rats for 15 consecutive days to examine the effect on ovarian hypertrophy. Sham-operated and hemicastrated control rats were administered a similar quantity of olive oil. The vaginal smear and body weight of the rats were recorded daily and rats were sacrificed on the Day 16. The ovary, uterus, kidney, adrenal, spleen, liver, lungs, heart, thymus, and thyroid were removed and weighed. The left ovary from each animal was serially sectioned and stained for histologic studies. The hemicastrated control rats revealed a significant increase in relative ovarian weight with 66.3% hypertrophy. Treatment with 700 and 800 mg/kg/day mancozeb revealed a decrease in ovarian hypertrophy with 28.2 and 22.8% hypertrophy, respectively. There was no significant change in the number of estrous cycles and duration of each phase of the estrous cycle with 500 mg/kg/day mancozeb. However, there was a decrease in the number of estrous cycles, duration of proestrus, estrus, and metestrus with concomitant significant increase in the duration of the diestrus phase with 600, 700, and 800 mg/kg/day mancozeb treatment. There was a significant decrease in the number of healthy follicles with concomitant increase in the number of atretic follicles at higher doses of mancozeb. There were no significant changes in the body and organ weight with 500, 600, 700, and 800 mg/kg/day of mancozeb. The levels of protein, glycogen, total lipid, phospholipid, and neutral lipid were elevated in the liver, uterus, and ovary after hemicastration. Protein, glycogen, total lipid, phospholipid, and neutral lipid were not significantly changed in the liver, uterus, and ovary after 500 mg/kg/day mancozeb. However, treatment with 600, 700, and 800 mg/kg/day mancozeb showed a significant decrease in the levels of protein, glycogen, total lipid, phospholipid, and neutral lipid in the liver, uterus, and ovary, with the exception of liver total lipid and uterine glycogen. In addition to the decrease in the compensatory ovarian hypertrophy, mancozeb treatment reduced the number of healthy follicles with a concomitant increase in the number of atretic follicles. This finding plus disruption of the estrous cycle may be due to a direct effect on the ovary or the hypothalamo-hypophysial-ovarian axis. PMID- 10825677 TI - A combined reproduction, neonatal development, and neurotoxicity study with 1,6 hexamethylene diisocyanate (HDI) in the rat. AB - 1,6-Hexamethylene diisocyanate (HDI), a chemical widely used in commercial polyurethane products, was evaluated in a combined reproductive/developmental/neurotoxicity study. Sprague-Dawley rats (n = 120; 15 per sex/dose group) were administered via whole-body inhalation exposure either 0, 0.005, 0.05, or 0.3 ppm HDI for 6 h/day during a 14-day premating phase, up to a 14-day mating phase, and a 21-day gestation phase. The dams and their litters were maintained for a 4-day lactation phase during which exposure to HDI was discontinued. Neurobehavioral testing (automated measures of activity and a functional observational battery) was conducted before exposure, after the premating phase, and before termination. Body weight and clinical observations were recorded throughout the study. Terminal examinations included a gross necropsy, hematology, and clinical chemistry. Tissues retained for microscopic examination included the reproductive organs, neural tissues, nasal turbinates (multiple sections), trachea, larynx, and lung. The animals were also evaluated for effects on mating, fertility, gestation length, litter size, pup sex ratio, and pup viability. In the 0.300 ppm dose group a statistically significant decrease in body weight was observed in the females on day 4 of the study. Also observed at this dose level, in both males and females, were microscopic alterations in the nasal cavity, primarily epithelial hyperplasia, squamous metaplasia, chronic-active inflammation, and more seriously, degeneration of the olfactory epithelium. Similar microscopic effects were also observed, albeit to a lesser extent, in the males and females of the 0.05 ppm dose level. No histopathologic effects were observed in the 0.005 ppm dose level. No effects on any reproductive or neurotoxicologic parameters, hematology, clinical chemistry, or any effects on pup growth and development were observed at any exposure level. PMID- 10825678 TI - Relationship between acrylamide reproductive and neurotoxicity in male rats. AB - To determine whether there is a relationship between the reproductive and neurotoxic effects of acrylamide monomer (AM), the first week of the study design of Sublet et al. ?14 was duplicated: Long-Evans male rats were gavaged with AM in water, 25/group, at 0, 5, 15, 30, 45, or 60 mg/kg/day for 5 days (days 1 through 5). On Day 8, males were paired overnight with untreated virgin females (1 : 1) in proestrus/estrus. On day 9, males were evaluated for forelimb and hindlimb grip strength. Five males/group were perfusion fixed, 20/group were used for andrologic assessment, and all were necropsied. Perfusion-fixed sciatic nerves were examined histologically. Sperm-positive females were examined for preimplantation and postimplantation loss at midpregnancy. At 15 to 60 mg/kg/day, males exhibited significantly reduced weight gain, reduced mating, fertility, and pregnancy indices by trend analysis (significant at 60 mg/kg/d by pairwise comparison), and increased postimplantation loss and dominant lethal factor, F(L)%, at 45 and 60 mg/kg/day. At 60 mg/kg/day, the sperm beat cross frequency was increased, with no significant effects on epididymal sperm motility or concentration, and hindlimb grip strength was decreased, with no pathologic lesions in sciatic nerves. Therefore, epididymal sperm, mating, and neurotoxic effects were observed at AM doses that also resulted in increased postimplantation loss, possibly by different mechanisms. PMID- 10825680 TI - Swelling of synovial joints - An anatomical, physiological and energy metabolical approach. AB - The direction and part of the rate of fluid filtration between synovial capillary (subscript capill) plasma and joint (subscript j) cavity fluid depends on the hydrostatic (P(capill)-P(j)) and colloid osmotic (COP(plasma)-COP(j)) pressure differences and on the osmotic efficiency (coefficient sigma) of 'colloids' across the barrier between plasma and synovial fluid. The rate of filtration also depends on capillary endothelial hydraulic conductance (L(p)) and surface area (A). Synovial fluid protein is removed by lymphatics, or split to be resorbed or used up by cells. Physiologists put together as equations the above, as well as other terms, all of these influenced by numerous factors. The equations offer a frame for a multifactorial approach to diseases associated with joint swelling and suggest, among others, that in the presence of weak generalized exsudation, synovial joints are loci minores resistentiae to swelling. The influence of joint cartilage energy demands on synovial blood flow may have been underrated. There are many accepted clinical, laboratory and morphological findings that fit this physiological frame that merits further clinical research. PMID- 10825681 TI - Effects of endothelin-1 on KCl-induced contractions of airway smooth muscles. AB - The objective of the study was to investigate the effect of endothelin-1 (ET-1) on KCl-induced contractions. Tracheal strips were obtained from 12 male rabbits, perfused with Krebs solution and aerated continuously by bubbling with a mixture of 95% O(2)-5% CO(2). Increasing concentrations of ET-1 (10(-12)-10(-7) M) additionally to KCl caused more contractions than KCl-induced (P<0.05). A total of 10(-7) M ET-1-induced 14.56% more contraction. Methylene blue (10(-5) M) decreased all the contractions induced from simultaneous use of KCl and ET-1. Methylene blue caused about 18% relaxation of contraction induced from KCl (80 mM) and ET-1 (10(-8) M). Consequently ET-1 increases the KCl-induced contractions of airway smooth muscles. This effect seem to mediate by a nitric oxide (NO) guanylate cyclase-cGMP pathway, in which methylene blue is inactivator of guanylate cyclase. PMID- 10825682 TI - Dose response of ethanol on antioxidant defense system of liver, lung, and kidney in rat. AB - This study investigated the alterations in levels of glutathione, lipid peroxidation, and antioxidant enzyme activity in the liver, lung, and kidney of rats treated with acute doses of ethanol. Male Fisher-344 rats were randomly divided into four groups, and were treated as follows: (1) vehicle (saline) control; (2) ethanol 2 g/kg, p.o.; (3) ethanol 4g/kg, p.o.; and (4) ethanol 6 g/kg, p.o. The animals were sacrificed 1 h after treatment, and tissues were isolated and analyzed. The hepatic GSH levels significantly decreased (73, 68, and 66% of control) due to ethanol ingestion at 2, 4, and 6g/kg, respectively. The hepatic GSH/GSSG ratio also decreased with increasing doses indicating stress response due to ethanol. The hepatic SOD activity significantly decreased (70, 75 and 71% of control) with graded doses of ethanol ingestion. The hepatic CAT/SOD and GSH-Px+CAT/SOD ratios significantly increased (147, 169 and 177% of control) and (140, 167 and 178% of control), respectively with increasing doses of ethanol. In the lung, graded doses of ethanol increased GSH-Px activity (120, 114 and 141% of control) and decreased GR activity (98, 89 and 89% of control), respectively. The MDA concentrations in the lung also increased after higher ethanol ingestion. Most of the antioxidant enzyme ratios increased with increasing doses of ethanol in the lung. In the kidney, GSH-Px activity increased (139, 119 and 151% of control), whereas GR activity decreased (84, 85 and 83% of control). GSH-Px/SOD and GSH-Px+CAT/SOD ratios increased whereas GR/GSH-Px ratio decreased after graded doses of ethanol. GSH levels in the kidney decreased after ethanol ingestion. MDA concentrations increased with increasing dose of ethanol in the kidney. These results showed the dose dependant and tissue specific changes in the antioxidant system after ethanol ingestion. Ethanol exerts oxidative stress on antioxidant systems of liver, lung and kidney in proportion to the amount of ethanol ingestion. PMID- 10825683 TI - Electrocorticographic factors associated with temporal lobe epileptogenicity. AB - Continuous subdural electrocorticographic (ECoG) monitoring was performed to test the hypothesis that human temporal lobe epileptogenicity, during long-term monitoring following antiepileptic drug (AED) withdrawal, regardless of the specific AED regimen, is dependent upon ECoG ictal onset and interhemispheric spread of epileptic activity. In 121 patients, ECoG parameters were analyzed for association with seizure frequency, a clinical measure of epileptogenicity. Significantly associated with increased seizure frequency were: ictal medial temporal lobe onset, absence of ictal frontal lobe desynchronization and short interhemispheric propagation time (IHPT). Seizure frequency during long-term ECoG monitoring was not predictive of post-operative seizure outcome. It is concluded that, following AED withdrawal, regardless of the specific AED regimen, increased seizure frequency is associated with medial temporal lobe ictal onset, short IHPT and absence of frontal lobe desynchronization. The results confirm the hypothesis that human temporal lobe epileptogenicity, after withdrawal, is dependent upon ECoG ictal onset and interhemispheric spread of epileptic activity. Future development of procedures which promote ECoG factors associated with increased seizure frequency following AED withdrawal might decrease duration of invasive long-term monitoring and improve efficiency for the pre-surgical selection of temporal lobectomy candidates. Intervention producing ictal frontal lobe desynchronization and increased IHPT might inhibit temporal lobe epileptogenicity and should be evaluated for therapeutic efficacy outside of the long-term monitoring context. PMID- 10825684 TI - Implication of ESR signals from ceruloplasmin (Cu(2+)) and transferrin (Fe(3+)) in pleural effusion of lung diseases. AB - Pleural effusions of seven lung cancer patients (mean age; 58) and seven non cancer patients (mean age; 49) were examined and Cu(2+) was measured in ceruloplasmin and Fe(3+) in transferrin signals by electron spin resonance (ESR) method. The variations of total Fe and Cu ions, ceruloplasmin and transferrin, proteins, neutrophil cell counts, LDH and nitrite/nitrate were also examined. The Cu(2+) peak was decreased and the Fe(3+) peak was increased in the cancer group. The interrelationship among Cu(2+), total Cu and ceruloplasmin, and among Fe(3+), total Fe and transferrin clarified that Cu(2+) and Fe(3+) are not a representative of ceruloplasmin and transferrin, respectively. The ratio of Cu(2+)/Fe(3+) in pleural effusion distinguished lung cancer from benign inflammation as a cause. The ratio of total Cu/total Fe measured by the chemical analysis method also distinguished these, but the ratio of ceruloplasmin/transferrin was unable to distinguish the cancer. In conclusion, the simple and rapid measurement of Cu(2+)/Fe(3+) by ESR effectively abstracts the variation of total ion concentrations caused by malignant disease. PMID- 10825685 TI - Is positive correlation between cortisol and met-enkephalin concentration in blood of women with breast cancer a reaction to stress before chemotherapy administration? AB - The role of stress in the course of neoplastic diseases has been emphasised over the past few years. Organism defence in form of increased release of hormones, corticosteroids and endogenous opioids should be assessed as a particular form of adaptation. Interactions between stress hormones and others which may influence the growth of breast cancer cells are possible. In this study, the concentration of cortisol and met-enkephalin vis-a-vis growth hormone, insulin-like growth factor-I, prolactin, estradiol, progesterone and melatonin in blood of women with breast cancer before the first cycle of supplementary chemotherapy was compared to that of healthy women. In a group of patients with breast cancer, before the first cycle of chemotherapy, high hypercortisolemia and positive correlation between cortisol and met-enkephalin was observed which may be a result of stress at the beginning of such treatment. A negative correlation between prolactin and met-enkephalin in this group can indirectly testify to essential participation of endogenous opioids in hormonal regulation and can be a response to stress caused by beginning chemotherapy. PMID- 10825686 TI - Induction of c-fos and interleukin-2 genes expression in the central nervous system following stressor stimuli. AB - An injection of 5% mustard in vegetable oil, an injection of vegetable oil and needle prick into a rat leg gastrocnemius muscle were selected as stressor stimuli. Expression of c-fos mRNA and interleukin-2 (IL-2) mRNA was measured using digoxigenin labeled cDNA probe. C-fos-like proteins were determined by an immunochemical method with anti-c-fos polyclonal antibodies. Thirty min after the injection of 5% mustard in vegetable oil, c-fos mRNA synthesis was shown in the sacral segments of the spinal cord. Many c-fos mRNA labeled cells were noted also in the hypothalamus (paraventricular nucleus, dorsomedial nucleus, and the lateral hypothalamic area), thalamus, hippocampus, nucleus caudatum, sensori motor zone of the brain cortex and the amygdaloid complex. Expression of c-fos like proteins was shown after the injection of vegetable oil and 5% mustard in vegetable oil (within 2 h) in the spinal cord structures, the maximum quantity of the labeled cells was found in the I and II sacral segments (in the superficial lamina of the gray matter of the dorsal horns in the ipsilateral side), the minimum in the IV and V lumbar segments (in the intermediate zone of the gray matter at the level of the central canal). When c-fos-like proteins expression were analyzed in the CNS, c-fos-labeled cells were detected in the same structures where c-fos mRNA was also shown. The IL-2 mRNA synthesis was shown within 2 h after the stimulus in the same brain structures as c-fos mRNA. It is always difficult to destinguish the effect of stress as sensory stimuli or as general stimuli. The c-fos immunoreactivity appeared mostly in the contralateral side of the hypothalamus, therefore maybe in our investigation the mechanism of sensory stimuli mostly results in the changes that have been obtained. The information of applied stimuli arrived in the spinal cord, and is transmitted to the brain where it activates certain brain structures inducing c-fos and IL-2 genes expression. The most important conclusion is that the used stress stimuli induced not only c-fos gene but also IL-2 gene expression in the brain. The synthesis of both mRNA takes place in the same brain structures. PMID- 10825687 TI - Superoxide dismutase activity and zinc and copper concentrations in Parkinson's disease. AB - Although several hypotheses are currently being investigated the cause of Parkinson's disease (PD) is still unknown. The aim of this study was to determine red cell copper/zinc-superoxide dismutase (Cu/Zn-SOD) activity and copper and zinc concentrations both in plasma and in red cell in PD. In this preliminary assay, 30 patients with PD the mean age of 64 were studied. Additionally, a second group of older individuals without PD mean age of 61, were recruited to the study. The patient group was compared with the other group according to their red cell Cu/Zn-SOD activities, and plasma and red cell copper, zinc concentrations. Red cell Cu/Zn-SOD activity was measured spectrophotometrically while plasma and red cell copper, zinc concentrations were determined by atomic absorption spectrophotometer. The results were analysed by 'Student t-test' statistically. The results showed that red cell Cu/Zn-SOD activities and red cell copper and zinc and also plasma copper concentrations of the PD patients increased compared to older individuals without PD. These findings suggested that possibility of oxidative stress in PD was reflected on the blood including the red cell and plasma parameters. PMID- 10825688 TI - Protection of gastric mucosal integrity by gelatin and simple proline-containing peptides. AB - We have observed that gelatin as feed supplement protected against ethanol induced mucosal damages in rats. Intraperitoneally given peptides PG and PGP had similar effects. The increase of mucosal integrity may be connected with the change of microcirculation: the peroral administration of PGP or gelatin given as a feed supplement decreased platelet aggregation. PMID- 10825689 TI - What makes plants different? Principles of extracellular matrix function in 'soft' plant tissues. AB - An overview of the biomechanic and morphogenetic function of the plant extracellular matrix (ECM) in its primary state is given. ECMs can play a pivotal role in cellular osmo- and volume-regulation, if they enclose the cell hermetically and constrain hydrostatic pressure evoked by osmotic gradients between the cell and its environment. From an engineering viewpoint, such cell walls turn cells into hydraulic machines, which establishes a crucial functional differences between cell walls and other cellular surface structures. Examples of such hydraulic machineries are discussed. The function of cell walls in the control of pressure, volume, and shape establishes constructional evolutionary constraints, which can explain aspects commonly considered typical of plants (sessility, autotrophy). In plants, 'cell division' by insertion of a new cell wall is a process of internal cytoplasmic differentiation. As such it differs fundamentally from cell separation during cytokinesis in animals, by leaving the coherence of the dividing protoplast basically intact. The resulting symplastic coherence appears more important for plant morphogenesis than histological structure; similar morphologies are realized on the basis of distinct tissue architectures in different plant taxa. The shape of a plant cell is determined by the shape its cell wall attains under multiaxial tensile stress. Consequently, the development of form in plants is achieved by a differential plastic deformation of the complex ECM in response to this multiaxial force (hydrostatic pressure). Current concepts of the regulation of these deformation processes are briefly evaluated. PMID- 10825690 TI - Natriuretic peptides in fish physiology. AB - Natriuretic peptides exist in the fishes as a family of structurally-related isohormones including atrial natriuretic peptide (ANP), C-type natriuretic peptide (CNP) and ventricular natriuretic peptide (VNP); to date, brain natriuretic peptide (or B-type natriuretic peptide, BNP) has not been definitively identified in the fishes. Based on nucleotide and amino acid sequence similarity, the natriuretic peptide family of isohormones may have evolved from a neuromodulatory, CNP-like brain peptide. The primary sites of synthesis for the circulating hormones are the heart and brain; additional extracardiac and extracranial sites, including the intestine, synthesize and release natriuretic peptides locally for paracrine regulation of various physiological functions. Membrane-bound, guanylyl cyclase-coupled natriuretic peptide receptors (A- and B-types) are generally implicated in mediating natriuretic peptide effects via the production of cyclic GMP as the intracellular messenger. C- and D-type natriuretic peptide receptors lacking the guanylyl cyclase domain may influence target cell function through G(i) protein-coupled inhibition of membrane adenylyl cyclase activity, and they likely also act as clearance receptors for circulating hormone. In the few systems examined using homologous or piscine reagents, differential receptor binding and tissue responsiveness to specific natriuretic peptide isohormones is demonstrated. Similar to their acute physiological effects in mammals, natriuretic peptides are vasorelaxant in all fishes examined. In contrast to mammals, where natriuretic peptides act through natriuresis and diuresis to bring about long-term reductions in blood volume and blood pressure, in fishes the primary action appears to be the extrusion of excess salt at the gills and rectal gland, and the limiting of drinking-coupled salt uptake by the alimentary system. In teleosts, both hypernatremia and hypervolemia are effective stimuli for cardiac secretion of natriuretic peptides; in the elasmobranchs, hypervolemia is the predominant physiological stimulus for secretion. Natriuretic peptides may be seawater adapting hormones with appropriate target organs including the gills, rectal gland, kidney, and intestine, with each regulated via, predominantly, either A- or B-type (or C- or D-type?) natriuretic peptide receptors. Natriuretic peptides act both directly on ion-transporting cells of osmoregulatory tissues, and indirectly through increased vascular flow to osmoregulatory tissues, through inhibition of drinking, and through effects on other endocrine systems. PMID- 10825691 TI - Effect of beta-antagonists on isoprenaline-induced secretion of fluid, amylase and protein by the parotid gland of the red kangaroo, Macropus rufus. AB - Selective and non-selective beta-adrenoceptor antagonists were used to block the increases in fluid, protein and amylase secretion caused by sympathomimetic stimulation of the parotid gland of red kangaroos during intracarotid infusion of isoprenaline. ICI118551 at antagonist/agonist ratios up to 300:1 caused increasing but incomplete blockade of fluid secretion, and protein/amylase release. Atenolol at antagonist/agonist ratios up to 300:1 was only marginally more potent than ICI118551 at blocking the fluid, protein and amylase responses. Propranolol at antagonist/agonist ratios of 30:1 was as effective at blocking fluid and protein secretion as the highest ratios of either atenolol or ICI118551. Simultaneous administration of atenolol (30:1) with ICI118551 (30:1) was not as potent as propranolol (30:1). Thus, the beta-adrenoceptor/s in the acini of the kangaroo parotid gland appear to have antagonist-binding affinities atypical of those found for eutherian tissues. The data are consistent with the gland possessing either a single anomalous beta-adrenoceptor or functional beta(2)-receptors in addition to the beta(1)-receptors which are characteristic of eutherian salivary glands. PMID- 10825692 TI - Energetics of lactation in domestic dog (Canis familiaris) breeds of two sizes. AB - The energetics of lactation was measured in two breeds of domestic dog during peak lactation. Labrador Retrievers (30 kg) had larger litter sizes than Miniature Schnauzers (6 kg). During the 7-day experimental period, Labrador pups increased more in mass than Schnauzer pups, both absolutely and relatively. Consequently, the energy demands of the litter, relative to maternal metabolism, were higher in Labradors than Schnauzers. Milk composition and gross efficiency of milk production were not significantly different between breeds and the costs of lactation were fuelled by increases in food intake. Metabolisable energy intake was higher than predicted in Labradors, but lower than predicted in Schnauzers. These patterns differ from interspecific expectations, which would predict larger animals to reproduce more slowly, have smaller litter sizes, and invest less energy in reproduction. PMID- 10825693 TI - Impact of hand-rearing on morphology and physiology of the capercaillie (Tetrao urogallus). AB - Morphological and physiological disparities between 20 captive and 11 wild capercaillies were determined. Birds, their pectoral and leg muscles, hearts, livers and gizzards were weighed. The length of small intestines and caeca were measured. Haemoglobin, haematocrit, glucose, triglycerides, total protein, uric acid and thyroid hormones as well as the cytochrome c-oxidase activity of the pectoral muscle and heart were determined. The glycogen and protein contents of pectoral and leg muscles and liver were analysed. Chemical composition (water, fat, protein, ash) of muscles and liver was determined. Captive males had heavier pectoral muscles than wild ones. The result was opposite in females. Wild birds had heavier hearts, livers, and gizzards, and also longer small intestines and caeca than captive birds. The cytochrome c-oxidase activity of pectoral muscle and heart was higher in wild than in hand-reared birds. The chemical composition of livers of wild birds differed significantly from that of hand-reared capercaillies. Plasma uric acid and T(4) concentrations were higher in captive than in wild birds. The observed differences in digestive system and liver can result in diminished ability of captive birds to utilise natural food nutrients. Decreased cytochrome c-oxidase activity of hand-reared birds can affect their takeoff and flying capacity and increase their vulnerability to predation. These facts may contribute to the low survival of hand-reared birds after release. PMID- 10825694 TI - Chloride transport by lobster hepatopancreas is facilitated by several anion antiport mechanisms. AB - Three anion antiporters have previously been demonstrated in lobster hepatopancreatic basolateral membrane vesicles (BLMV) to perform vital physiological functions in the crustacean. Cl(-) was shown to be transported by all three of the documented antiporters. The stilbene, 4-acetamido-4' isothiocyanostilbene-2,2'-disulfonic acid, also known as SITS, strongly inhibited Cl(-)/SO(4)(2-), Cl(-)/oxalate(2-) and Cl(-)/HCO(3)(-) exchange. It was concluded that Cl(-) could be transported by different modes of the documented existing anion antiporters in the lobster hepatopancreatic BLMV. PMID- 10825695 TI - Expression of 70 kDa heat shock proteins in antarctic and New Zealand notothenioid fish. AB - The cold and constant water temperature of the Southern Ocean surrounding Antarctica provides a natural laboratory to address questions of temperature adaptation in marine organisms. In this study, endogenous levels and the number of isoforms of the 70 kDa heat shock protein multigene family (hsp70) of Antarctic and cold temperate notothenioid fishes were determined by SDS polyacrylamide gel electrophoresis and Western blotting. Tissues from three Antarctic Trematomus congeners had significantly lower levels of 70 kDa Hsp isoforms than their temperate confamilial from New Zealand waters. However, these two thermally disparate sets of fish did not differ in number or pattern of 70 kDa Hsp isoforms expressed under normal physiological conditions. Additionally, levels of 70 kDa Hsp isoforms in specimens of one Antarctic species, Trematomus bernacchii, acclimated to 4 degrees C were significantly higher than non acclimated conspecifics, indicating a direct effect of temperature on Hsp expression in this species. This study shows that constitutive expression of some members of the 70 kDa Hsp multigene family have been maintained, despite the absence of environmental heat stress for at least 2.5 million years. PMID- 10825696 TI - The hormonal regulation of premigratory fat deposition and winter fattening in red-winged blackbirds. AB - Glucagon is a highly potent lipolytic agent in birds and a candidate for regulating premigratory and winter fattening. The seasonal role of glucagon in fat metabolism was determined by monitoring plasma glucagon, fatty acids and glucose in two groups of red-winged blackbirds; one group exposed to outside environmental conditions (September to May) and a second group maintained at summer conditions with respect to day length and temperature. The results of this investigation demonstrate significantly lower plasma glucagon (480.1 pg/ml) in birds exposed to outdoor conditions than in birds maintained at summer conditions (734.6 pg/ml) during September/October. The data are consistent with the view that low plasma glucagon in outdoor birds ensures the preservation of fat stores for autumn migration. Lower plasma free fatty acid (FFA) levels (0.35 mEq/l) in outdoor birds (vs. 0.54 mEq/l in indoor birds) in autumn may reflect the rapid transport of FFA to adipose tissue for lipogenesis resulting in a steady increase in body weight from September to January. The sharp decline in plasma FFA in indoor birds from 0.54 mEq/l in September/October to 0.28 mEq/l in January/February may be attributed to a marked decrease in food consumption, rather than a dramatic change in the rate of lipid transport from blood to muscle or adipose tissue. Glucagon injections caused a 600% increase in plasma FFA and a more modest (50%) increase in plasma glucose. This confirms the major role of glucagon in fat mobilization. Its lipolytic effects, however, can vary seasonally by way of down regulation of glucagon receptors. Down regulation of glucagon receptors in adipose tissue and the associated reduced sensitivity of adipocytes to the lipolytic action of glucagon would account for the progressive increase in weight of the birds throughout November/December when plasma glucagon levels were significantly higher (578.9 pg/ml) in outdoor birds as compared to indoor birds (436.9 pg/ml). Lower plasma glucagon levels (405.5 pg/ml) in outdoor birds in January/February (vs. 638.6 pg/ml in indoor birds) may reflect the same physiological conditions prevailing in September/October favoring the preservation of fat stores. PMID- 10825697 TI - Heart rate within male crayfish: social interactions and effects of 5-HT. AB - Behaviors, such as those that establish dominant and subordinate social status, are thought to be driven by various neuromodulators and hormones. In crustaceans, the level of serotonin (5-HT) in the hemolymph is correlated with degree of aggressiveness. The crustacean heart is neurogenic and is modulated by neural secretion of 5-HT in the hemolymph, which bathes the cardiac tissue. We discuss and present the results of measuring heart rate (HR) of crayfish during interactions, as an indication of their state of excitability. HR is the result of multiple influences: a cocktail of hormones and modulators. HR was monitored during the periods in which crayfish established aggressive and submissive social status, during sham injections, and following injections of various doses of 5 HT. Crayfish, during an interaction to establish social status, can increase HR. Both the aggressive and submissive crayfish can dampen their HR within seconds during a pause in the interaction, while still posturing in an aggressive or submissive state. Injections of 5-HT to obtain systemic levels of approximately 100 nM-10 microM increase HR substantially for hours. This suggests that aggressive interactions and the establishment of a dominant posture may not be related to large increases in the free concentrations of 5-HT within the circulating hemolymph, since a sustained HR is not observed in aggressive animals. Instead, the results may demonstrate that inhibitory cardiac regulation is present in the aggressors during interactions and that a regulator is possibly 5-HT. PMID- 10825698 TI - Potential role of pyrroline 5-carboxylate in regulation of collagen biosynthesis in cultured human skin fibroblasts. AB - Although insulin-like growth factor-I (IGF-I) is known as an important stimulator of collagen biosynthesis in collagen-producing cells, the mechanism and endpoints by which it regulate the process remain largely unknown. Serum of acutely fasted rats contained reduced amount of IGF-I (72+/-16 ng/ml) and showed about 75% reduced ability to stimulate collagen and DNA synthesis in confluent human skin fibroblasts in comparison to the effect of control rat serum (IGF-I, 168+/-19 ng/ml). An addition of IGF-I (at least 40 ng/ml) to fasted rat serum restored its mitogenic activity but could not restore its ability to stimulate collagen biosynthesis to control values during 24 h of incubation. However, when the cells were incubated in fasted rat serum supplemented with 40 ng/ml of IGF-I for 48 h, collagen biosynthesis was restored to control values. It suggests that the stimulatory role of IGF-I in collagen biosynthesis undergo indirectly. We considered pyrroline-5-carboxylate (P5C) as a candidate to play a direct role in this process. Since IGF-I and P5C are known to be decreased in serum of fasted rats it seems that the action of IGF-I on collagen biosynthesis may involve participation of P5C. We have found that serum of fasted rats (showing low level of P5C) supplemented with 1 mmol/l P5C induced collagen biosynthesis in confluent human skin fibroblasts during 24 h to control values. Supporting evidence comes from the experiment showing stimulatory action of P5C on collagen biosynthesis in fibroblasts cultured in serum-free medium. Our results postulate potential role of P5C in regulation of collagen biosynthesis and indicate participation of this molecule in the pathway of IGF-I action in this process. PMID- 10825699 TI - Comparison of calcium storage between a Baikalian gastropod and holarctic relatives. AB - In Lake Baikal, extremely thin shells are reported as a typical feature of endemic gastropods. This statement derived only from observations; no experimental data were available up to now. Therefore, we quantitatively investigated the calcium distribution in the endemic prosobranch gastropod Benedictia baicalensis and compared the results with those of Lithoglyphus naticoides, a near relative, non-endemic, palaearctic species. The shell of the endemic mollusc B. baicalensis consists of 94.9+/-26.0 microg Ca(2+)/microl animal volume (n=43), and in L. naticoides 865.0+/-271.5 microg Ca(2+)/microl (n=10). Calcium contents in the tissue of B. baicalensis vary between different sampling stations and different sampling dates (from 9.4+/-5.1 (n=33) to 20.5+/ 8.4 microg Ca(2+)/mg dry weight DW (n=16)) and are only 1/5-1/10 compared to L. naticoides (88.5+/-39.1 microg Ca(2+)/mg DW (n=9)). But the values for hemolymph calcium concentration and osmolality in both species are identical (B. baicalensis: osmolality: 84.4+/-5.3 mosm/kg (n=40); hemolymph calcium concentration: 4.6+/-1.7 mmol/l (n=40). L. naticoides: osmolality: 85.0+/-2.0 mosm/kg (n=8); hemolymph calcium concentration: 5.2+/-5.0 mmol/l (n=40).). This is the first experimental study demonstrating, that - besides a similar hemolymph ionic composition - the Baikalian species is characterized by significantly lower calcium storage in shell and tissue than the nearly related non-endemic species. PMID- 10825700 TI - Brain chirps: spectrographic signatures of epileptic seizures. AB - OBJECTIVE: A chirp is a brief signal within which the frequency content changes rapidly. Spectrographic chirps are found in signals produced from many biological and physical phenomena. In radar and sonar engineering, signals with chirps are used to localize direction and range to the signal source. Although characteristic frequency changes during epileptic seizures have long been observed, the correlation with chirps and chirp technology seems never to have been made. METHODS: We analyzed 19404 s (1870 s of which were from 43 seizures) of intracranially (subdural and depth electrode) recorded digital EEG from 6 patients for the presence of spectral chirps. Matched filters were constructed from methods in routine use in non-medical signal processing applications. RESULTS: We found that chirps are very sensitive detectors of seizures (83%), and highly specific as markers (no false positive detections). The feasibility of using spectral chirps as matched filters was demonstrated. CONCLUSIONS: Chirps are highly specific and sensitive spectrographic signatures of epileptic seizure activity. In addition, chirps may serve as templates for matched filter design to detect seizures, and as such, can demonstrate localization and propagation of seizures from an epileptic focus. PMID- 10825701 TI - Oculomotor phenomena in petit-mal. AB - OBJECTIVE: To determine the temporal relationship among behavior, eye movements and respiration during absence seizures. METHOD: This included simultaneous videorecording of a patient's face, EEG and respirogram, in 5 patients with absence seizures. Absence seizures were defined as a sudden lapse of consciousness with impairment of mental functions. Oculomotor phenomena consisted of: eye fluttering, eye deviation (conjugate lateral or upward deflection), and eye-opening. RESULTS: Oculomotor phenomena began 1 2/3-3 s after the onset of the electrographic discharge and ended before the discharge. The most frequent events were eye-opening, stare with or without palpebral myoclonias, and tonic or clonic upward movements. Downward movement or convergence was not observed as well as head version. Respiratory changes (apnea) are the last events to occur. CONCLUSIONS: absence seizures were characterized by the onset of a 3/s spike-slow wave discharge followed by oculomotor phenomena and respiratory changes. The time course suggests that epileptic discharges precede and then involve oculomotor and respiratory brainstem centers. PMID- 10825702 TI - Contribution of motor cortex in generation of evoked spikes in patients with benign rolandic epilepsy. AB - OBJECTIVES: Among the different kinds of rolandic epilepsy there is a form of benign epilepsy with centrotemporal spikes (BECT) presenting the peculiar characteristic of evoking rolandic paroxysmal activity, characterized by a spike followed by a slow wave, using electrical stimulation of the fingers. METHODS: We evaluated 7 patients suffering from BECT presenting evoked scalp activity by electrical stimulation of the fingers of the hand. Electrical stimulation was performed using a pair of ring electrodes applied to the thumb. The motor evoked potentials (MEPs) were elicited in hand muscles by transcranial magnetic stimulation (TMS) and were conditioned by the same electrical digital stimulation producing the evoked spikes at interstimulus intervals ranging from 10 to 200 ms. RESULTS: Digital stimulation in epileptic patients produced an increase in MEP amplitude substantially above the normal ranges. MEP facilitation showed a time course overlapping the ascending phase and peak of the evoked spike, whereas no significant MEP changes were found during the early positive peak and the descending phase of the spike, or during the following slow wave. CONCLUSIONS: Several considerations support the hypothesis that the short-lasting M1 facilitation is related to the spread of an abnormal hypersynchronous discharge of the S1 neurones to functionally related motor areas via cortico-cortical connections. PMID- 10825703 TI - The organization of quick corrections within a two-joint synergy in conditions of unexpected blocking and release of a fast movement. AB - OBJECTIVE: Within this study, we tested a hypothesis that common organization of central commands to the elbow and the wrist joints within a two-joint synergy is associated with a similar organization of pre-programmed corrections to unexpected changes in the external conditions. DESIGN AND METHODS: The subjects (n = 7) performed series of very fast movements or isometric contractions against a pad at the level of the palm or at the level of the forearm. Some trials within a series of movements were unexpectedly blocked at the initial position leading to an isometric contraction, while some trials within a series of isometric contractions were unexpectedly released leading to a movement. Movement kinematic and electromyographic (EMG) patterns were analyzed. RESULTS: In cases of unexpectedly blocked or unexpectedly released trials, differences in the EMG patterns between perturbed and unperturbed trials were seen at latencies between 50 and 80 ms. Two patterns were observed representing interindividual differences among subjects. One included reciprocal changes in the flexor-extensor pairs controlling both joints. The other pattern included reciprocal changes in the wrist flexor-extensor pair and unidirectional changes in the EMGs of biceps and triceps. The patterns were reproducible within each individual subject across tasks and conditions. In particular, the same pattern in the wrist flexor extensor pair was seen when the pad was applied to the palm and when it was applied to the forearm, despite the fact that early wrist joint deviations were in opposite directions. CONCLUSIONS: It is argued that the observed early EMG changes may be unrelated to local joint kinematics and stretch reflexes, and represent consequences of control patterns for fast corrective movements that are organized with respect to the motion of the endpoint of the limb similarly to the organization of voluntary movements. Within this framework, the organization of joints of a multi-joint limb into a synergy implies a simultaneous, automatic organization of pre-programmed reactions into a similarly organized synergy. PMID- 10825704 TI - Voluntary teeth clenching facilitates human motor system excitability. AB - OBJECTIVE: Voluntary contraction of the teeth is a common maneuver used to facilitate peripheral monosynaptic reflexes. It was the aim of this study to determine the site along the neuraxis where this effect occurs. METHODS: Focal transcranial magnetic stimulation (TMS) was used to measure recruitment curves, motor thresholds and intracortical inhibition and facilitation from the right first dorsal interosseus (FDI) and tibialis anterior (TA) muscles in seven normal volunteers. Changes in excitability in subcortical structures during teeth clenching were studied using F waves, H reflexes, and brainstem magnetic stimulation. RESULTS: Recruitment curves of FDI and TA showed significant facilitation during voluntary teeth clenching indicating an overall enhancement in the motor system excitability. Teeth clenching additionally resulted in decreased intracortical inhibition in the FDI but not in TA, pointing to an intracortical site of enhancement for the hand. Motor evoked potentials (MEPs) following stimulation at the brainstem level and F waves in FDI and soleus H reflex amplitude were also facilitated by teeth clenching, indicating a subcortical site for this effect for the upper and lower extremity. M wave amplitudes did not change. CONCLUSIONS: The teeth clenching maneuver had a similar facilitatory effect on upper and lower extremities. Cortical and subcortical sites contribute to this effect in a hand muscle while only subcortical sites were identified in this facilitatory effect on the lower extremity. PMID- 10825705 TI - Motor cortical dysfunction disclosed by single and double magnetic stimulation in patients with fibromyalgia. AB - OBJECTIVE: To investigate the motor cortex by single and double magnetic stimulation, in patients with fibromyalgia. METHODS: Thirteen patients with fibromyalgia and 13 age-matched healthy subjects were examined. We evaluated, in both limbs, motor evoked potential (MEP) latency and amplitude and the MCA/MPA ratio, i.e. MEP cortical amplitude (MCA) /maximal peripheral amplitude of the M response (MPA), the central conduction time (TCC) and the length of the silent period (SP). With double magnetic stimulation, different time intervals between shocks were used: with delays between shocks of 4, 25, 55 and 85 ms, the intensities of the conditioning shock were 80% the relaxed threshold. With delays between shocks of 55, 85, 100, 155, 200, 255 and 355 ms, the intensities of the conditioning shocks were set at 150% the relaxed threshold. In all cases, the intensity of the test shock was 150% the relaxed threshold. The results were also compared with those obtained in 5 women affected by rheumatoid arthritis (RA). RESULTS: As compared to control, the cortical relaxed threshold was enhanced on both sides and limbs (P<0.05). The cortical silent period recorded with single magnetic stimulation was reduced in the upper limbs (P = 2.7x10(-11)) and lower limbs (both sides P = 3.6x10(-5)). The other parameters investigated were normal. With double magnetic stimulation, facilitatory phenomena were absent in fibromyalgic patients and the inhibitory responses recorded with a delay of 155 ms were reduced (P = 0.0052). No significant differences were noted between FM and RA patients. CONCLUSION: This study demonstrated motor cortical dysfunction in patients with fibromyalgia involving excitatory and inhibitory mechanisms. This indicates motor cortical involvement and supports the hypothesis of aberrant central pain mechanisms. The absence of differences between FM and RA suggest that the lesions were not specific and could be related to chronic pain disorders within the central nervous system. PMID- 10825706 TI - Effects of low-frequency transcranial magnetic stimulation on motor excitability and basic motor behavior. AB - OBJECTIVE: To explore effects of low-frequency repetitive transcranial magnetic stimulation (rTMS) of the primary motor cortex (M1) on motor excitability and basic motor behavior in humans. DESIGN AND METHODS: Seven normal volunteers underwent 1 Hz rTMS of the hand representation of the right M1 for 15 min at an intensity of 115% of the individual resting motor threshold. The effects of rTMS on motor excitability were assessed by monitoring changes in individual resting motor threshold and input-output curves of motor evoked potentials (MEPs) in the flexor pollicis brevis, first dorsal interosseus, abductor digiti minimi and biceps brachii muscles. Changes in basic motor behavior were studied by measuring maximal and mean peak force and peak accelerations of thumb flexions and abductions of the fifth finger before and after rTMS. RESULTS: rTMS produced a significant increase in resting motor threshold and a significant suppression of MEP input-output curves that persisted for 30 min. The suppressing effect was restricted to the hand motor representation which was the prime target of the stimulation procedure, and there were no significant effects on the biceps representation. Peak force and peak acceleration were not affected while the motor representations of muscles involved in the behavioral measurements were significantly suppressed by rTMS. CONCLUSIONS: Low-frequency rTMS of M1 transiently depresses motor excitability but this does not affect basic motor behavior. This is relevant for the therapeutic use of low-frequency rTMS in disorders with abnormal cortical excitability. PMID- 10825707 TI - Effect of antagonistic voluntary contraction on motor responses in the forearm. AB - OBJECTIVE: We investigated the effects of voluntary contraction of agonist and antagonist muscles on motor evoked potentials (MEP) and on myoelectric activities in the target (agonist) muscle following transcranial magnetic stimulation (TMS). METHODS: The left extensor carpi radialis (ECR) and flexor carpi radialis (FCR) muscles were studied in 16 healthy subjects. H reflexes, MEP induced by TMS, and background electromyographic (EMG) activity were recorded using surface electrodes at rest and during voluntary contraction of either agonist or antagonist muscles. RESULTS: Voluntary contraction of antagonist muscles (at 10% of maximum contraction) enhanced the amplitudes of MEP for both muscles. The H reflex of the FCR muscle was inhibited by contraction (10% of maximum) of the ECR muscle. Background EMG activity did not differ between H-reflex trials and TMS trials. Enhancement of MEP amplitudes and background EMG activity during voluntary antagonist contraction was comparable in the two muscles. Appearance rate of MEP recorded by needle electrodes in response to subthreshold TMS was increased by antagonistic voluntary contraction. CONCLUSION: Facilitation occurs during voluntary contraction of antagonist muscles. Differences between the effects of voluntary contraction of the ECR muscle for the MEP and the H reflex of the FCR suggest that cortical facilitatory spread occurs between agonist and antagonist muscles. PMID- 10825708 TI - Quantitative measures of spasticity in post-stroke patients. AB - OBJECTIVE: Quantitative evaluation of muscle tone in post-stroke patients; correlation of biomechanical indices with conventional clinical scales and neurophysiological measures; characterization of passive and neural components of muscle tone. METHODS: Mechanical stretches of the wrist flexor muscles of 53 post stroke patients were imposed by means of a torque motor at constant speed. Patients were clinically studied using the Ashworth scale for spasticity and the Medical Research Council score for residual muscle strength. The neurophysiological measures were Hoffmann reflex latency, Hmax/Mmax ratio, stretch reflex threshold speed (SRTS), stretch reflex (SR) latency and area, passive (ISI) and total (TSI) stiffness indices. RESULTS: Hmax/Mmax ratio, SR area, ISI and TSI values were significantly higher in patients, while SRTS was significantly lower. TSI, SRTS and SR area were highly correlated to the Ashworth score. CONCLUSIONS: This EMG-biomechanical technique allows an objective evaluation of changes in muscle tone in post-stroke patients, providing easily measurable, quantitative indices of muscle stiffness. The linear distribution of these measures is particularly indicated for monitoring changes induced by treatment. The apparatus seems suitable to characterize neural stiffness, while difficulties were found in isolating the passive components, because of the occurrence of tonic EMG activity in most spastic patients. PMID- 10825709 TI - Influence of GAA expansion size and disease duration on central nervous system impairment in Friedreich's ataxia: contribution to the understanding of the pathophysiology of the disease. AB - OBJECTIVE: To verify if GAA expansion size could account for the severity of the central nervous system involvement in Friedreich's ataxia (FA). METHODS: Retrospective study of 52 FA patients (mean age 26.9+/-12.1 years; mean disease duration 10.6+/-7.6 years) homozygous for GAA expansion. Median nerve somatosensory evoked potentials (SSEPs) were available in 36 FA patients, upper limb motor evoked potentials (MEPs) to transcranial magnetic stimulation in 32, brainstem auditory evoked potentials (BAEPs) in 24, and visual evoked potentials (VEPs) in 34. N20, P100, MEP amplitude, SSEP and MEP central conduction time (CCT and CMCT), P100 latency and I-III and I-V interpeak latency, and a BAEP abnormality score were correlated with disease duration and GAA expansion size on the shorter (GAA1) and larger (GAA2) allele in each pair. RESULTS: The GAA1 size inversely correlated with the N20 amplitude (r = -0.49; P<0. 01). Disease duration directly correlated with CMCT (r = 0.57; P<0.01) and BAEP score (r = 0.61; P<0.01) and inversely with MEP (r = -0.40; P<0.05) and P100 amplitude (r = 0.39; P<0.05). CONCLUSIONS: Our data suggest that central somatosensory pathway involvement in FA is mainly determined by GAA1 expansion size. Vice versa, degeneration of pyramidal tracts, auditory and visual pathways seems to be a continuing process during the life of FA patients. PMID- 10825711 TI - The cause of slowed forearm median conduction velocity in carpal tunnel syndrome. AB - OBJECTIVES: Attempting to answer a debate concerning the etiopathogenesis of the decreased forearm median motor conduction velocity (FMMCV), we tried to use proximal stimulation at the wrist, elbow, mid-arm and axillary regions to determine segmental median motor conduction velocity (MMCV). We also correlated the FMMCV with median motor distal latency (MMDL) and compound muscle action potential (CMAP) amplitudes of the abductor pollicis brevis (APB) muscle in order to assess whether the conduction block of large myelinating fibers or retrograde axonal atrophy was the major cause of the decreased FMMCV. BACKGROUND: The cause of the decreased FMMCV resulting from either the conduction block of the large myelinating fibers at the wrist or distal compression with retrograde axonal atrophy remains an unresolved issue at the moment. Animal models have supported the hypothesis that the retrograde axonal atrophy might also occur in humans. Other authors believe the standard FMMCV is calculated by subtracting the distal latency which may not represent an exact assessment of FMMCV but rather the velocity of small fibers that persist through the carpal tunnel. SUBJECTS AND METHODS: Patients with the clinical symptoms and signs of carpal tunnel syndrome (CTS) confirmed using standard electrodiagnosis were included. The patients were arbitrarily divided into two groups based on the FMMCV, one with reduced FMMCV (n = 20, FMMCV < 50 m/s) and the other with normal FMMCV (n = 35, FMMCV> or =50 m/s). Age-matched volunteers served as controls. We explored motor conduction proximally at wrist, elbow, mid-arm and axillary stimulation, and recorded at the APB muscles. Based on the latency differences, we calculated the FMMCV, distal arm MMCV (DAMMCV) and proximal arm MMCV (PAMMCV), and compared the conduction velocity (CV) differences of DAMMCV-FMMCV, PAMMCV-FMMCV and PAMMCV-DAMMCV in the two patient groups and the control. Furthermore, we correlated FMMCV with MMDL and CMAP amplitudes of APB muscle because MMDL and CMAP amplitudes might reflect the integrity of the large myelinating fibers. RESULTS: CMAP amplitudes of APB muscle at wrist stimulation and MMDL were not correlated with FMMCV in either of the two patient groups; however, the CMAP amplitude was markedly decreased and MMDL was significantly prolonged when compared with normal controls. The significant increase of CV gradient of DAMMCV-FMMCV and PAMMCV-FMMCV without an equal increase of CV gradient of PAMMCV-DAMMCV only occurred in the reduced FMMCV patient group, suggesting that the conduction block is not the primary cause. The CV gradient of DAMMCV-FMMCV and PAMMCV-DAMMCV did not show any significant difference between patients with the normal FMMCV and the control group. CONCLUSION: The retrograde axonal atrophy, not selective damage of the large fibers at the wrist, was the direct cause of the decreased FMMCV. PMID- 10825710 TI - Amyotrophic lateral sclerosis versus cervical spondylotic myelopathy: a study using transcranial magnetic stimulation with recordings from the trapezius and limb muscles. AB - OBJECTIVE: We report an electrophysiological method to differentiate amyotrophic lateral sclerosis (ALS) from cervical spondylotic myelopathy (CSM). METHODS: Motor evoked potentials (MEPs) by transcranial magnetic stimulation were investigated in patients with ALS (n=10) and CSM (n=9). In addition to limb MEPs using the triple stimulation technique (TST) at upper limbs, MEPs recorded from trapezius muscles were compared with those obtained from 23 normal subjects. The parameters studied were: central motor conduction time, amplitude ratio and, for the trapezius, the interside asymmetry. RESULTS: Whereas limb MEPs were abnormal in most ALS and CSM patients (17/19), trapezius MEPs were abnormal in all ALS patients, and normal in 8 out of 9 CSM patients. CONCLUSION: Recording of trapezius MEPs is a valuable addition to the limb MEPs study, since it distinguishes ALS from SCM in most patients. PMID- 10825712 TI - ERP correlates of response inhibition to elemental and configural stimuli in a negative patterning task. AB - OBJECTIVE: The present experiment examined the ERP correlates of response inhibition to elemental and configural Nogo stimuli in a Go/Nogo task. DESIGN AND METHODS: Event-related potentials (ERPs) were recorded while 8 subjects completed a visual Go/Nogo task. Nogo stimuli required the inhibition of a response to stimuli that differed from Go stimuli (A+, B+) either on the basis of each of two physical features (elemental Nogo stimuli; CD-) or on the basis of the conjunction of features represented in the Go stimuli (configural Nogo stimuli; AB-). Behavioural data and ERP component measures (amplitude and latency) were analysed using analysis of variance. RESULTS: An enhanced N2 component and an enhanced fronto-centrally distributed P3 component were elicited following elemental Nogo stimuli relative to Go stimuli, consistent with a number of studies examining ERPs during Go/Nogo tasks. In contrast, an enhanced late frontal negative/parietal positive slow wave was elicited following configural Nogo stimuli relative to Go stimuli. CONCLUSIONS: These results cast doubt on the interpretation of the N2 enhancement as reflecting response inhibition processes per se. The pattern of results was interpreted as providing support for the unique cue model of learning rather than the configural model of learning and was discussed in the context of a recent model of executive functioning. PMID- 10825713 TI - Duration and frequency mismatch negativity in schizophrenia. AB - OBJECTIVES: The aim of the present study was to elucidate the reasons for apparent inconsistencies in the schizophrenia literature with respect to the mismatch negativity (MMN) waveform of the event-related potential (ERP). While most previous research has shown that MMN is reduced in schizophrenia, there are a small number of studies reporting that frequency MMN is not reduced. METHODS: We recorded ERPs to auditory stimuli with different frequencies and durations from patients with schizophrenia (N = 14) and control subjects (N = 17) of similar age and sex. MMNs to small but discriminable frequency deviants were contrasted with large frequency deviants and duration deviants. RESULTS: Only the MMN to duration deviants was significantly reduced in patients, although there was evidence of a similar trend for large frequency deviants. CONCLUSIONS: The results together with a review of the frequency MMN literature suggest that there are 3 variables which are important in determining whether patients exhibit a reduced MMN to frequency deviants: deviant probability, degree of deviance and interstimulus interval. The results also indicated that patients with schizophrenia may have particular deficits in processing the temporal properties of auditory stimuli. This finding has implications for the pathophysiology of the disorder as time-dependent processing is reliant on the integrity of an extensive network of brain areas consisting of auditory cortex, areas of pre-frontal cortex, the basal ganglia and cerebellum. PMID- 10825714 TI - Quantitative sensory testing: effect of site and pressure on vibration thresholds. AB - OBJECTIVE: To determine the effect of different sites and locally applied pressure on vibration thresholds. METHODS: Vibration thresholds were compared in 47 normal volunteers at 3 sites of the index finger (pulp, dorsum of the middle phalanx and nail) and at two sites of the great toe (dorsum of the proximal phalanx and nail). The effect of local pressure (30, 50 and 100 g/1.22 cm(2)) were compared in 41 subjects at the dorsum of the middle phalanx of the index finger and the proximal phalanx of the great toe. RESULTS: The hand was more sensitive than the foot for vibration. There were no significant differences in vibration thresholds at different sites of the index finger and different sites of the great toe. The pulp of the index finger yielded the least inter-individual variation. Testing under 30 and 50 g/1.22 cm(2) of pressure yielded equal vibration thresholds. Vibration threshold was higher when tested under 100 g/1.22 cm(2) at the index finger but not the great toe. This difference was small and clinically negligible. CONCLUSION: Testing of vibration thresholds in normal subjects can be adequately conducted at several sites of the index finger and the great toe. The test can be adequately done under low pressure of 30-50 g/1.22 cm(2). PMID- 10825715 TI - Maturation of near-field and far-field somatosensory evoked potentials after median nerve stimulation in children under 4 years of age. AB - OBJECTIVES: The maturation of subcortical SEPs in young children. METHODS: Median nerve SEPs were recorded during sleep in 42 subjects aged 0-48 months. Active electrodes were at the ipsilateral Erb's point, the lower and upper dorsal neck, and the frontal and contralateral centroparietal scalp; reference electrodes were at the contralateral Erb's point, the ipsilateral earlobe and the frontal scalp; bandpass was 10-3000 Hz. The peaks were labelled by their latencies in adults. RESULTS: The peak latencies of N9 (brachial plexus potential) decreased exponentially with age during the first year, but increased with height thereafter. The interpeak latencies (IPLs) N9-N11, which measure conduction between brachial plexus and dorsal column, decreased with age (linear regression). The IPLs N11-P13 and N11-N13b, which measure conduction between the dorsal column and approximately the cervico-medullary junction, did not change across this age range. The IPLs N13a-N20, N13b-N20 and P13-N20, which measure central conduction, showed negative exponential regressions with rapidly decreasing latencies during the first year of life and slowly decreasing latencies thereafter. CONCLUSIONS: Maturation of the peripheral segments of the somatosensory pathway progresses more rapidly than that of the central segments. The maturation of central conduction is not completed within the first 4 years of age. Our maturational data may serve as a reference source for subsequent developmental and clinical studies. PMID- 10825717 TI - Differential interaction of somatosensory inputs in the human primary sensory cortex: a magnetoencephalographic study. AB - OBJECTIVE: Somatosensory evoked magnetic fields (SEFs) were recorded to investigate the interaction of the somatosensory inputs using the modality of electrical finger stimulation in 6 normal subjects. METHODS: Electrical stimuli were given to the index (II), middle (III) or little (V) fingers individually, and also to pairs of either the II and III simultaneously, or the II and V simultaneously. The interaction ratio (IR) was calculated as the ratio of the SEF amplitude by simultaneous two-finger stimulation to the arithmetically summed SEF amplitudes of two individual-finger stimulations. RESULTS: SEFs showed 3 major components: N22m, P30m and P60m. The N22m and P60m revealed a clear somatotopic organization in the primary sensory cortex (S1) in the sequence of II, III and V, while the P30m showed a cluster with medial location compared with N22m and P60m in S1. The N22m had a significantly greater IR in II and III stimulation compared to that in II and V stimulation. The P60m also showed a similar trend in the IR but was greater than that of N22m. In contrast, the IR in P30m showed no such tendency. CONCLUSION: The interaction of S1 was most influenced when adjacent receptive fields were activated in the modality of electrical finger stimulation. Our results were consistent with the concept that the Brodmann's areas in S1 which produce the 3 components of the SEFs have different functional organization. PMID- 10825716 TI - Intra- and postoperative factors determining neurological complications after surgery under deep hypothermic circulatory arrest: a retrospective somatosensory evoked potential study. AB - OBJECTIVES: Intraoperative median nerve SEP monitoring uses the disappearance of cortical and brain stem activities as the criterion to determine that brain cooling is sufficient in deep hypothermic circulatory arrest (CA) surgery. This study presents the results of a retrospective SEP analysis of intraoperative events that engendered neurological complications. METHODS: Median nerve SEP monitoring was performed on 58 consecutive patients who underwent surgery under deep hypothermic CA. The monitoring was retrospectively analyzed and compared with intraoperative events, and postoperative symptoms. RESULTS: Intraoperative SEP abnormalities were observed in 9 out of the 16 patients who presented neurological complications. Among the 7 others, the abnormalities either were present preoperatively (n = 2), occurred postoperatively (n = 3) or only involved the lower limbs (n = 2). Although the mere inspection of the intraoperative SEPs was insufficient to determine the origin of the alterations, their comparison with intraoperative events facilitated the identification of hemodynamic factors in 7 cases and embolism in two cases. There were no patients in whom CA per se caused neurological complications. CONCLUSIONS: Intraoperative SEP monitoring helps identify intraoperative events responsible for neurological complications and prevent these in subsequent procedures. However, the neurological complications of deep hypothermic CA can also be due to pre- or postoperative factors that escape the domain of intraoperative monitoring. PMID- 10825718 TI - Sources of cortical responses to painful CO(2) laser skin stimulation of the hand and foot in the human brain. AB - OBJECTIVES: To investigate whether the same dipolar model could explain the scalp CO(2) laser evoked potential (LEP) distribution after either hand or foot skin stimulation. METHODS: LEPs were recorded in 14 healthy subjects after hand and foot skin stimulation and brain electrical source analysis of responses obtained in each individual was performed. RESULTS: A 5 dipolar sources model explained the scalp LEP topography after both hand and foot stimulation. In particular, we showed that the co-ordinates of the two earliest activated dipoles were compatible with source locations in the upper bank of the Sylvian fissure on both sides. These sources did not change their location when the stimulation site was moved from the upper to the lower limb. The other 3 dipoles of our model were activated in the late LEP latency range with a biphasic profile and a location compatible with activation of the cingulate gyrus and deep temporo-insular structures. CONCLUSIONS: The dipolar model previously proposed for the hand stimulation LEPs can also satisfactorily explain the LEP distribution obtained after foot stimulation. The earliest activated Sylvian dipolar sources did not change their location when the upper or lower limb was stimulated, as expected from the close projections of hand and foot in the second somatosensory area. No source in the primary somatosensory area was necessary to model the scalp topography of LEPs to hand and foot stimulation. PMID- 10825719 TI - On the processing of spatial frequencies as revealed by evoked-potential source modeling. AB - OBJECTIVES AND METHODS: Visually evoked potentials (VEPs) are known to be sensitive to spatial frequency, especially in the time range between 50 and 100 ms post-stimulus. In two experiments we localized the cortical activity elicited by stimuli of varying spatial frequency in scalp-recorded brain potentials, using multi-electrode recordings and dipole-source analysis. RESULTS: Low spatial frequencies (<1 c/d) activated relatively lateral occipital areas, the orientation of the neural ensembles involved being predominantly perpendicular to the scalp surface. In contrast, high spatial frequencies (>4 c/d) induced activation of more medial occipital areas with the predominant orientation of the sources being much more parallel to the scalp surface. Furthermore, at about 100 ms latency the lateral-occipital response to low spatial frequencies was stronger in the right hemisphere; no such asymmetry was found for the responses to the high spatial frequencies. These findings were consistent across varying recording conditions, individual subjects, subject populations, stimulus characteristics (grating orientation, grating vs. checkerboard), and task conditions (active vs. passive). CONCLUSION: The results indicate that there are differences in sensitivity to specific spatial frequencies between primary and secondary visual areas, as well as between the right and the left hemispheres. PMID- 10825720 TI - Comparison of visual and auditory evoked cortical potentials in migraine patients between attacks. AB - OBJECTIVE: As both habituation of pattern reversal visual evoked potentials (PR VEP) (Schoenen J, Wang W, Albert A, Delwaide PJ. Potentiation instead of habituation characterizes visual evoked potentials in migraine patients between attacks. Eur J Neurol 1995;2:115-122) and intensity dependence of auditory evoked cortical potentials (IDAP) (Wang W, Timsit-Berthier M, Schoenen J. Intensity dependence of auditory evoked potentials in migraine: an indication of cortical potentiation and low serotonergic neurotransmission? Neurology 1996;46:1404-1409) were found abnormal in migraine between attacks, we have searched for intraindividual correlations between both tests in 59 migraine patients (22 with aura [MA], 37 without aura [MO]) and in 23 healthy volunteers (HV). METHODS: Amplitude change of the PR-VEP N1-P1 was measured between the 1st and 5th block of 50 sequential averagings during continuous stimulation at 3.1 Hz. IDAP was computed from N1-P2 amplitudes of 100 averagings during stimulations at 40, 50, 60 and 70 dB SL. Amplitude-stimulus intensity function (ASF) slopes and amplitude changes between 40 and 70 dB were calculated. MO and MA differed from HV in PR VEP amplitude change (P=0.007) and IDAP slope (P = 0.0004). RESULTS: There was no significant correlation between VEP amplitude changes and IDAP slopes, nor between the latter two and attack frequency or disease duration. A negative correlation was found between the amplitude of the first block of averaged responses and potentiation of VEP in all subject groups (P = 0.03) as well as between the amplitude of the auditory evoked potential, at 40 dB, and the percentage of amplitude increase between 40 and 70 dB in MO (P = 0.004) and MA (P = 0.007). ASF slopes and 40 dB amplitudes were significantly correlated only in the MA group (P = 0.002). These results confirm the interictal deficit of habituation in cortical processing of repetitive visual and auditory information in migraine. Since there is no intraindividual correlation between the cortical responses to these sensory modalities they are complementary tools for the study of migraine and may help to identify subgroups of patients with distinct pathophysiological mechanisms. CONCLUSIONS: The strong negative correlation between the initial amplitude of evoked potentials and their amplitude increase during subsequent averaging confirms that the response potentiation in migraine is likely to be due to a reduced preactivation level of sensory cortices. PMID- 10825721 TI - Vigilance stages and performance in OSAS patients in a monotonous reaction time task. AB - OBJECTIVES: To develop improved methods for objective assessment of daytime vigilance. This is important in the diagnosis and therapy control of sleep disorders associated with excessive daytime sleepiness (EDS). METHODS: Twenty-one patients with EDS due to obstructive sleep apnea were recorded during a daytime 90 min reaction time (RT) test in a monotonous situation. Two EEG, two EOG and a submental EMG channel were recorded simultaneously. The recordings were divided into short, stationary segments of variable length (0.5-2 s) and classified into 7 stages using our previously described system, which includes additional stages for drowsiness. RESULTS: The duration of RT was linearly correlated to the vigilance state (P<0.05). The appearance of slow eye movements (SEM) was more consistently related to performance impairment than EEG changes. CONCLUSIONS: Our system can provide a better tracking of vigilance changes than the standardized sleep stage scoring. A 1-2 h test is useful in the assessment of the performance of a subject suffering from EDS. We found that SEMs indicate more sensitive and consistent EDS-related performance impairment than changes in EEG activity. PMID- 10825722 TI - The Human Genome Project - obstacles versus opportunities. PMID- 10825724 TI - Mouse model of spinal muscular atrophy. PMID- 10825723 TI - New insights into factors that affect HIV-1 transmission. PMID- 10825726 TI - Beyond uHTS: ridiculously HTS? PMID- 10825725 TI - Funding for knowledge-sake - Letter. PMID- 10825727 TI - Can you stand the heat of today's drug discovery? PMID- 10825729 TI - Applications of NMR in drug discovery. AB - In the half-century since its discovery, nuclear magnetic resonance (NMR) has become the single most powerful form of spectroscopy in both chemistry and structural biology. The dramatic technical advances over the past 10-15 years, which continue apace, have markedly increased the range of applications for NMR in the study of protein-ligand interactions. These form the basis for its most exciting uses in the drug discovery process, which range from the simple identification of whether a compound (or a component of a mixture) binds to a given protein, through to the determination of the full three-dimensional structure of the complex, with all the information this yields for structure based drug design. PMID- 10825730 TI - Respiratory syncytial virus: recent progress towards the discovery of effective prophylactic and therapeutic agents. AB - Although respiratory syncytial virus (RSV) was discovered in 1955, the burden associated with this infectious agent on all population groups is only now beginning to be fully appreciated. The successful launch of the humanized monoclonal antibody Synagis (developed by MedImmune, Gaithersburg, MD, USA), as a prophylactic in September 1998 has helped to heighten awareness of the extent of mortality and morbidity associated with annual RSV epidemics. Small, drug-like molecules that would provide the clinician with effective and conveniently administered prophylactic and therapeutic agents for the prevention and treatment of RSV have not yet advanced into clinical studies. This review will summarize recent developments in the area of RSV drug discovery and development. PMID- 10825731 TI - In vitro display technologies - new tools for drug discovery. AB - Over the past decade, several ligand discovery techniques have been developed that mimic the process of natural evolution. Phage display technology is the most established of these methods and has been applied to numerous technological problems including the discovery of novel drugs. More recently, some new display technologies have emerged which, unlike phage display, operate entirely in vitro and have concomitant advantages. This review describes this new generation of display technologies and indicates how they might fit into the modern drug discovery process. PMID- 10825732 TI - Monitor: molecules and profiles. AB - Monitor provides an insight into the latest developments in drug discovery through brief synopses of recent presentations and publications together with expert commentaries on the latest technologies. There are two sections: Molecules summarizes the chemistry and the pharmacological significance and biological relevance of new molecules reported in the literature and on the conference scene; Profiles offers commentary on promising lines of research, emerging molecular targets, novel technology, advances in synthetic and separation techniques and legislative issues. PMID- 10825733 TI - Combinatorial chemistry. PMID- 10825734 TI - Bioinformatics. PMID- 10825735 TI - Perforin: more than just an effector molecule. PMID- 10825737 TI - Autoimmunity can benefit self-maintenance. AB - Autoimmunity is usually considered only as a cause of disease; nevertheless, human T-cell repertoires are filled naturally with autoimmune lymphocytes. Here, we review evidence that autoimmune T cells can help heal damaged tissues, indicating that natural autoimmunity could also be a cause of health. PMID- 10825736 TI - Jekyll and Hyde: the transformation of HLA-B27. PMID- 10825738 TI - Both self and non-inherited maternal HLA antigens influence the immune response. AB - Recent studies suggest that exposure of the fetus and newborn to non-inherited maternal major histocompatibility complex HLA antigens (NIMA) has a life-long effect on allograft recognition that could influence tolerance of organ grafts. NIMA also appear to influence disease susceptibility. Here, Jon van Rood and Frans Claas discuss evidence that three HLA haplotypes, those inherited from the parents plus NIMA, shape the immune response. PMID- 10825739 TI - Interleukin 16: implications for CD4 functions and HIV-1 progression. AB - In this article, David Center and colleagues clarify the controversies that have emerged over the unique structure of interleukin 16 and its anti-HIV-1 activity. Interleukin 16 is a ligand for CD4, and this implies CD4 acts as a sentinel receptor that can switch CD4+ T cells between immune and inflammatory functions. PMID- 10825740 TI - Neural immunoregulation: emerging roles for nerves in immune homeostasis and disease. AB - In this review, James Downing and Jaleel Miyan outline emerging evidence for neural mechanisms that contribute to specific categories of host defence. Involvement of direct innervation in the adaptive control of immunological responses complements an established view of neuroendocrine-immune modulation. The challenge remains to understand the integrative and homeostatic functions of 'hardwiring' of peripheral immune effector sites, its bearing on disorder and potential for therapeutic modification. PMID- 10825741 TI - A new primate model for multiple sclerosis in the common marmoset. AB - Experimental autoimmune encephalomyelitis (EAE) in outbred marmoset monkeys (Callithrix jacchus) is a recently developed nonhuman primate model of multiple sclerosis. Here, Bert 't Hart and colleagues compare this model to EAE in rhesus monkeys, highlighting autoimmune mechanisms in CNS inflammation and demyelination, including the role of major histocompatibility complex restriction and preclinical evaluation of innovative immunotherapies. PMID- 10825743 TI - Reply to Balasa and Sarvetnick. PMID- 10825742 TI - Molecular insights into the immunopathogenesis of follicular lymphoma. AB - Follicular lymphoma is caused by the transformation of a germinal-center-derived B cell with a t(14;18) chromosomal translocation. The distribution of somatic mutations within immunoglobulin genes indicates that follicular-lymphoma cells can interact with antigen. In addition, nonimmunoglobulin genes such as BCL6 seem to undergo somatic hypermutation. Here, Kostas Stamatopoulos and colleagues relate the molecular data about immunoglobulin genes and the protooncogenes BCL2 and BCL6 to the pathogenesis and evolution of follicular lymphoma. PMID- 10825744 TI - Reply to infante, krolick and wall PMID- 10825746 TI - Reply to saoudi et al PMID- 10825745 TI - Is pathogenic humoral autoimmunity a Th1 response? PMID- 10825747 TI - Interplanetary crew dose rates for the August 1972 solar particle event. AB - Parsons, J. L. and Townsend, L. W. Interplanetary Crew Dose Rates for the August 1972 Solar Particle Event. Using the coupled neutron-proton space radiation transport computer code (BRYNTRN), estimates of dose rates of protons in the skin, ocular lens and bone marrow, behind various thicknesses of aluminum shielding, for crews on space missions outside the Earth's magnetosphere, are made for the large solar particle event (SPE) of August 1972. Overall, the August 1972 dose rates are significantly higher than those estimated for any of the events that occurred in August-December 1989. The dose rates in the August 1972 SPE are not low dose rates as specified by the major national and international advisory bodies and committees. PMID- 10825748 TI - The relative biological effectiveness of low doses of 14 MeV neutrons in steady state murine spermatogenesis as determined by flow cytometry. AB - The relative biological effectiveness of 14 MeV neutrons in the low-dose range < or =1 Gy has been determined in differentiating and differentiated spermatogonia. Male NMRI mice were exposed to single doses of 2 cGy to 3 Gy of (60)Co gamma rays or neutrons. The ratios of testicular S-phase cells, 4c primary spermatocytes, and elongated spermatids were quantified by DNA flow cytometry 2 to 70 days after irradiation and were found to decrease. Histological samples and testis weight were analyzed in parallel. Doses of 2-5 cGy neutrons and 10-50 cGy gamma rays significantly (P<0.05) decreased the proportions of S-phase cells, spermatocytes and elongated spermatids at 4, 14 and 28 days postirradiation. For S-phase cells, the biphasic shape of the cell survival curves was described with a D(50) of 5 cGy neutrons. The D(50) for (60)Co gamma rays and the relative biological effectiveness could not be determined. The relative biological effectiveness of neutrons at 50% reductions of testis weight, primary spermatocytes, and elongated spermatids were 2.5, 10.0 and 6.1, respectively. This in vivo assay is interesting because of its sensitivity at dose ranges that are relevant for exposures in the environment, the workplace and radiotherapy. PMID- 10825749 TI - Measuring the spectrum of mutation induced by nitrogen ions and protons in the human-hamster hybrid cell line A(L)C. AB - Astronauts can be exposed to charged particles, including protons, alpha particles and heavier ions, during space flights. Therefore, studying the biological effectiveness of these sparsely and densely ionizing radiations is important to understanding the potential health effects for astronauts. We evaluated the mutagenic effectiveness of sparsely ionizing 55 MeV protons and densely ionizing 32 MeV/nucleon nitrogen ions using cells of two human-hamster cell lines, A(L) and A(L)C. We have previously characterized a spectrum of mutations, including megabase deletions, in human chromosome 11, the sole human chromosome in the human-hamster hybrid cell lines A(L)C and A(L). CD59(-) mutants have lost expression of a human cell surface antigen encoded by the CD59 gene located at 11p13. Deletion of genes located on the tip of the short arm of 11 (11p15.5) is lethal to the A(L) hybrid, so that CD59 mutants that lose the entire chromosome 11 die and escape detection. In contrast, deletion of the 11p15.5 region is not lethal in the hybrid A(L)C, allowing for the detection of chromosome loss or other chromosomal mutations involving 11p15.5. The 55 MeV protons and 32 MeV/nucleon nitrogen ions were each about 10 times more mutagenic per unit dose at the CD59 locus in A(L)C cells than in A(L) cells. In the case of nitrogen ions, the mutations observed in A(L)C cells were predominantly due to chromosome loss events or 11p deletions, often containing a breakpoint in the pericentromeric region. The increase in the CD59(-) mutant fraction for A(L)C cells exposed to protons was associated with either translocation of portions of 11q onto a hamster chromosome, or discontinuous or "skipping" mutations. We demonstrate here that A(L)C cells are a powerful tool that will aid in the understanding of the mutagenic effects of different types of ionizing radiation. PMID- 10825750 TI - Chromosome translocations in turtles: a biomarker in a sentinel animal for ecological dosimetry. AB - Nonhuman organisms are being exposed to ionizing radiations at radionuclide contaminated sites around the world. Direct methods are seldom available for measuring biologically relevant doses received by these organisms. Here we extend biological dosimetry techniques, which are much better developed for humans and a few other mammalian species, to a nonmammalian species. Turtles were chosen because a long-lived animal would best serve the need for low-level, chronic exposure conditions. We chose the yellow-bellied slider turtle (Trachemys scripta), which is known to have a maximum life span of at least 22 years. As reported elsewhere, we first isolated an embryonic fibroblast cell line and constructed whole-chromosome-specific DNA libraries for chromosome 1 by microdissection and PCR. A FISH painting probe was prepared and used to establish a dose-response curve for ionizing radiation-induced chromosome interchange aberrations in turtle fibroblasts. This was compared to the dose response for human fibroblasts treated under similar conditions in our laboratory. With respect to induction of chromosome interchange aberrations, human fibroblasts were approximately 1.7 times more sensitive than the T. scripta fibroblasts. To the extent that symmetrical interchanges are persistent over long periods, this approach could eventually provide a measure of the integrated lifetime dose these organisms receive from radionuclides in their environment and give a measure of the extent of relevant genetic damage over that time. PMID- 10825751 TI - Analysis of blood lymphocyte subsets in children living around Chernobyl exposed long-term to low doses of cesium-137 and various doses of iodine-131. AB - Epidemiological studies have found that children living around Chernobyl have rates of respiratory tract illness that are higher than those seen in the area before the Chernobyl accident. The present study investigates the possible effects of radiation exposure on the composition of peripheral blood lymphocyte subsets in children living around Chernobyl. Two hundred nineteen healthy children and children suffering from recurrent respiratory diseases aged 6-14 years who received both low doses of radiation to the whole body from (137)Cs and various doses of radiation to the thyroid from (131)I as fallout from the accident were assessed 5 (1991) and 8-10 years (1994-1996) after the accident. A total of 148 healthy children and children suffering from recurrent respiratory diseases living in noncontaminated areas were also evaluated as controls. Children with recurrent respiratory diseases who lived around Chernobyl had a significantly lower percentage of T cells and a higher percentage of NK cells compared to control children with recurrent respiratory diseases during the study period. In contrast to the findings in 1991, a significant decrease in the percentage of helper-inducer cells was observed in children with recurrent respiratory diseases in 1994-1996. In contrast to 1991, there is a positive correlation between the percentage of helper-inducer cells, the helper inducer/cytotoxic-suppressor cell ratio, and the dose of radiation to the thyroid of healthy children from (131)I in 1994-1996. There was a positive correlation between the dose of radiation to the thyroid from (131)I and the percentage of helper-inducer cells in children with recurrent respiratory diseases 5 years (1991) after the accident. Further, the dose of radiation to the thyroid from (131)I correlated negatively with the percentage of T and B cells and positively with the percentage of NK cells in children with recurrent respiratory diseases 8 10 years (1994-1996) after the accident. These results raise the possibility that long-term exposure to low doses of (137)Cs may have altered the composition of the T-cell subsets and NK cells in children with recurrent respiratory diseases. The differences in the composition of the peripheral blood lymphocyte subsets between healthy children and those with recurrent respiratory diseases may be attributed to long-term low-dose exposure of the whole body to radiation from (137)Cs and exposure of the thyroid to radiation from (131)I subsequent to the Chernobyl accident. PMID- 10825752 TI - Biological response to ionizing radiation in mouse embryo fibroblasts with a targeted disruption of the DNA polymerase beta gene. AB - Base excision repair (BER) is carried out by two distinct pathways in mammalian cells, one dependent on DNA polymerase beta (Polb) and the other on proliferating cell nuclear antigen (Pcna). We studied whether the Polb-dependent pathway plays an important role in BER in vivo after exposure to ionizing radiation. For this purpose, we used mouse embryo fibroblasts derived from wild-type and Polb gene knockout littermates. Both cell lines had essentially the same clonogenic cell survival and low levels of apoptosis as determined by a colony formation assay and by a change in mitochondrial membrane potential, respectively. No significant cleavage of protein kinase C delta (Pkcd) in vivo, which is a substrate for caspase 3, was detected, and intact Pkcd was retained in both cell lines for at least 72 h after irradiation. Similar significant increases in caspase 3-like activities as measured by Asp-Glu-Val-Asp (DEVD) cleaving activity in vitro were observed in both cell lines after irradiation. Radiation induced cell cycle arrest in the form of a G(2)-phase block, and G(2)/M-phase fractions reached a peak approximately 10 h after irradiation and decreased thereafter with a similar time course in both cell lines. Similar levels of chromatin-bound Pcna were observed immediately after irradiation in non-S-phase cells of both cell lines and disappeared by 4 h after irradiation. We conclude that the deficiency in Polb does not have a significant influence on the radiation responses of these cells. Together with evidence accumulated in vitro, these results strongly support the idea that the Pcna-dependent pathway predominantly acts in BER of radiation induced DNA damage in vivo. PMID- 10825753 TI - Glucose-6-phosphate dehydrogenase and the oxidative pentose phosphate cycle protect cells against apoptosis induced by low doses of ionizing radiation. AB - The initial and rate-limiting enzyme of the oxidative pentose phosphate shunt, glucose-6-phosphate dehydrogenase (G6PD), is inhibited by NADPH and stimulated by NADP(+). Hence, under normal growth conditions, where NADPH levels exceed NADP(+) levels by as much as 100-fold, the activity of the pentose phosphate cycle is extremely low. However, during oxidant stress, pentose phosphate cycle activity can increase by as much as 200-fold over basal levels, to maintain the cytosolic reducing environment. G6PD-deficient (G6PD(-)) cell lines are sensitive to toxicity induced by chemical oxidants and ionizing radiation. Compared to wild type CHO cells, enhanced sensitivity to ionizing radiation was observed for G6PD( ) cells exposed to single-dose or fractionated radiation. Fitting the single-dose radiation response data to the linear-quadratic model of radiation-induced cytotoxicity, we found that the G6PD(-) cells exhibited a significant enhancement in the alpha component of radiation-induced cell killing, while the values obtained for the beta component were similar in both the G6PD(-) and wild-type CHO cell lines. Here we report that the enhanced alpha component of radiation induced cell killing is associated with a significant increase in the incidence of ionizing radiation-induced apoptosis in the G6PD(-) cells. These data suggest that G6PD and the oxidative pentose phosphate shunt protect cells from ionizing radiation-induced cell killing by limiting the incidence of radiation-induced apoptosis. The sensitivity to radiation-induced apoptosis was lost when the cDNA for wild-type G6PD was transfected into the G6PD(-) cell lines. Depleting GSH with l-BSO enhanced apoptosis of K1 cells while having no effect in the G6PD(-) cell line PMID- 10825754 TI - Induction and rejoining of DNA double-strand breaks in bladder tumor cells. AB - The induction and rejoining of radiation-induced double-strand breaks (DSBs) in cells of six bladder tumor cell lines (T24, UM-UC-3, TCC-SUP, RT112, J82, HT1376) were measured using the neutral comet assay. Radiation dose-response curves (0-60 Gy) showed damage (measured as mean tail moment) for five of the cell lines in the same rank order as cell survival (measured over 0-10 Gy), with the least damage in the most radioresistant cell line. Damage induction correlated well with clonogenic survival at high doses (SF10) for all six cell lines. At the clinically relevant dose of 2 Gy, correlation was good for four cell lines but poor for two (TCC-SUP and T24). The rejoining process had a fast and slow component for all cell lines. The rate of these two components of DNA repair did not correlate with cell survival. However, the time taken to reduce the amount of DNA damage to preirradiated control levels correlated positively with cell survival at 10 Gy but not 2 Gy; radioresistant cells rejoined the induced DSBs to preirradiation control levels more quickly than the radiosensitive cells. Although the results show good correlation between SF10 and DSBs for all six cell lines, the lack of correlation with SF2 for TCC-SUP and T24 cells would suggest that a predictive test should be carried out at the clinically relevant dose. At present the neutral comet assay cannot achieve this. PMID- 10825755 TI - Comparison of repair of DNA double-strand breaks in identical sequences in primary human fibroblast and immortal hamster-human hybrid cells harboring a single copy of human chromosome 11. AB - We have optimized a pulsed-field gel electrophoresis assay that measures induction and repair of double-strand breaks (DSBs) in specific regions of the genome (Lobrich et al., Proc. Natl. Acad. Sci. USA 92, 12050-12054, 1995). The increased sensitivity resulting from these improvements makes it possible to analyze the size distribution of broken DNA molecules immediately after the introduction of DSBs and after repair incubation. This analysis shows that the distribution of broken DNA pieces after exposure to sparsely ionizing radiation is consistent with the distribution expected from randomly induced DSBs. It is apparent from the distribution of rejoined DNA pieces after repair incubation that DNA ends continue to rejoin between 3 and 24 h postirradiation and that some of these rejoining events are in fact misrejoining events, since novel restriction fragments both larger and smaller than the original fragment are generated after repair. This improved assay was also used to study the kinetics of DSB rejoining and the extent of misrejoining in identical DNA sequences in human GM38 cells and human-hamster hybrid A(L) cells containing a single human chromosome 11. Despite the numerous differences between these cells, which include species and tissue of origin, levels of TP53, expression of telomerase, and the presence or absence of a homologous chromosome for the restriction fragments examined, the kinetics of rejoining of radiation-induced DSBs and the extent of misrejoining were similar in the two cell lines when studied in the G(1) phase of the cell cycle. Furthermore, DSBs were removed from the single-copy human chromosome in the hamster A(L) cells with similar kinetics and misrejoining frequency as at a locus on this hybrid's CHO chromosomes. PMID- 10825756 TI - Radiation-induced heat-labile sites that convert into DNA double-strand breaks. AB - The yield of DNA double-strand breaks (DSBs) in SV40 DNA irradiated in aqueous solution was found to increase by more than a factor of two as a result of postirradiation incubation of the DNA at 50 degrees C and pH 8.0 for 24 h. This is in agreement with data from studies performed at 37 degrees C that were published previously. Importantly, similar results were also obtained from irradiation of mammalian DNA in agarose plugs. These results suggest that heat labile sites within locally multiply damaged sites are produced by radiation and are subsequently transformed into DSBs. Since incubation at 50 degrees C is typically employed for lysis of cells in commonly used pulsed-field gel assays for detection of DSBs in mammalian cells, the possibility that heat-labile sites are present in irradiated cells was also studied. An increase in the apparent number of DSBs as a function of lysis time at 50 degrees C was found with kinetics that was similar to that for irradiated DNA, although the magnitude of the increase was smaller. This suggests that heat-labile sites are also formed in the cell. If this is the case, a proportion of DSBs measured by the pulsed-field gel assays may occur during the lysis step and may not be present in the cell as breaks but as heat-labile sites. It is suggested that such sites consist mainly of heat-labile sugar lesions within locally multiply damaged sites. Comparing rejoining of DSBs measured with short and long lysis procedure indicates that the heat-labile sites are repaired with fast kinetics in comparison with repair of the bulk of DSBs. PMID- 10825757 TI - Radioprotective thiolamines WR-1065 and WR-33278 selectively denature nonhistone nuclear proteins. AB - Differential scanning calorimetry was used to study the interactions of nuclei isolated from Chinese hamster V79 cells with the radioprotector WR-1065, other thiol compounds, and polyamines. Differential scanning calorimetry monitors denaturation of macromolecules and resolves the major nuclear components (e.g. constrained and relaxed DNA, nucleosome core, and nuclear matrix) of intact nuclei on the basis of thermal stability. WR-1065 treatment (0.5-10 mM) of isolated nuclei led to the irreversible denaturation of nuclear proteins, a fraction of which are nuclear matrix proteins. Denaturation of 50% of the total nonhistone nuclear protein content of isolated nuclei occurred after exposure to 4.7 mM WR-1065 for 20 min at 23 degrees C. In addition, a 22% increase in the insoluble protein content of nuclei isolated from V79 cells that had been treated with 4 mM WR-1065 for 30 min at 37 degrees C was observed, indicating that WR 1065-induced protein denaturation occurs not only in isolated nuclei but also in the nuclei of intact cells. From the extent of the increase in insoluble protein in the nucleus, protein denaturation by WR-1065 is expected to contribute to drug toxicity at concentrations greater than approximately 4 mM. WR-33278, the disulfide form of WR-1065, was approximately twice as effective as the free thiol at denaturing nuclear proteins. The proposed mechanism for nucleoprotein denaturation is through direct interactions with protein cysteine groups with the formation of destabilizing protein-WR-1065 disulfides. In comparison to its effect on nuclear proteins in isolated nuclei, WR-1065 had only a very small effect on non-nuclear proteins of whole cells, isolated nuclear matrix, or the thiol-rich Ca(2+)ATPase of sarcoplasmic reticulum, indicating that WR-1065 can effectively denature protein only inside an intact nucleus, probably due to the increased concentration of the positively charged drug in the vicinity of DNA. PMID- 10825758 TI - Free radical formation in X-irradiated crystals of 2'-deoxycytidine hydrochloride. Electron magnetic resonance studies at 10 K. AB - Single crystals of deoxycytidine hydrochloride (CdR.HCl) have been X-irradiated at 10 K with doses up to about 150 kGy and studied using 24 GHz (K-band) EPR, ENDOR and FSE spectroscopy. In this system, the cytosine base is protonated at the N3 position. Nine different radicals were characterized and identified. Three of these are ascribed to three versions of the one-electron reduced species, probably differing in their protonation state. Radicals formed by net hydrogen addition to the cytosine C5 and C6 positions were observed at 10 K. The hydrogen abstraction radical at the deoxyribose C1' position most probably results from initial oxidation of the base. The remaining radical species are all localized to the sugar moiety, representing products formed by net hydrogen abstraction from three of the five available carbons of the deoxyribose sugar. The lack of base centered oxidation products as well as the structures of the one-electron reduced species is rationalized by considering the specific proton donor-acceptor properties of this crystalline lattice in comparison with similar systems. PMID- 10825759 TI - Electron paramagnetic resonance investigation of sucrose irradiated with heavy ions. AB - We investigated the potential use of sucrose to estimate linear energy transfer (LET) for heavy-ion irradiation. We also made a quantitative comparison between heavy-ion and gamma irradiation in terms of spin concentration. Heavy-ion irradiation of sucrose produces stable free radicals. Based on the electron paramagnetic resonance (EPR) spectra obtained, the stable sucrose radicals are the same among helium ions, carbon ions and gamma rays. The EPR spectrum was approximately 70 G wide and was composed of several hyperfine structures. The total spin concentration obtained after the heavy-ion irradiation increased linearly as the absorbed dose increased and decreased logarithmically as LET increased. Production of the spin concentration of helium ions was two times more dependent on LET than that for carbon-ion irradiation. The empirical relationships obtained imply that LET at a certain dose can be determined by the spin concentration. Furthermore, the results of gamma irradiation of deuterated sucrose suggest that one of the persistent radicals is a carbon-centered radical. PMID- 10825760 TI - Survival of a mixture of cells of variable linear-quadratic sensitivity to radiation. AB - The experimentally observed survival of a heterogeneous mixture of cells, each component of which obeys a different linear-quadratic survival response to ionizing radiation, is examined. It is shown that the survival relationship for the mixed population approaches a linear-quadratic form for low doses. The linear parameter of the low-dose relationship approached is equal to the average of the distribution of values of the linear parameter (alpha(i)) of the various components of the mixture. The quadratic parameter of the low-dose relationship approached is equal to the average of the distribution of values of the quadratic parameter (beta(i)) of the various components of the mixture minus one-half the variance of the distribution of the values of alpha(i). A numerical example of the survival expected for an exponentially growing population of Chinese hamster V79 cells is presented. From this it can be appreciated that the apparent value of the alpha and beta parameters obtained by fitting an experimentally obtained survival curve will depend on the range of doses over which survival is determined. The apparent value of beta is decreased at higher doses, producing a straightening of the survival curve to approach the exponential decrease in survival commonly observed for the terminal high-dose portion of survival curves. Apparent exponential survival at high doses is not inconsistent with linear quadratic survival and may not indicate a multitarget or other mechanism of cell killing. PMID- 10825761 TI - Hemochromatosis heterozygotes may constitute a radiation-sensitive subpopulation. AB - A primary mechanism of radiation-induced DNA damage is by generation of free radicals. Chronically increased oxidative stress from elevated levels of iron in the body may increase radiation sensitivity by decreasing cellular oxygen radical scavenging capability. Hemochromatosis heterozygotes have elevated body iron. Low level radiation sensitization by iron may be particularly pertinent for risk of breast cancer. Since 10% of the population appears to be heterozygous for the hemochromatosis gene, a radiosensitizing effect would have pervasive implications. PMID- 10825763 TI - Visible stents: all that glitters...is it gold? PMID- 10825762 TI - Multicenter registry to evaluate the efficacy of the NIROYAL stent in de novo or restenotic coronary stenosis. AB - The purpose of this registry was to document the safety and efficacy of elective deployment of the NIROYAL stent (Boston Scientific SCIMED, Maple Grove, Minnesota) in coronary arteries. This was a prospective, multi-center international registry. NIROYAL stents (9, 16, 25 and 32 mm-long) were manually crimped onto coronary balloons and deployed in de novo or restenotic lesions in 165 male and female patients with angina pectoris and a reference vessel diameter of 2.0-4.5 mm. Stent deployment was successful in 98.8% of the lesions. Mean percent diameter stenosis decreased from 83.5+/-10.8% to 2.7+/- 6.2% after intervention. The interventional procedure of the treatment site was successful in 97.6% of patients. At six-month clinical follow-up, 88.5% had event-free survival [i.e., did not experience a major adverse cardiac event (MACE)] and 95.2% had no anginal symptoms. Furthermore, 87.3% of the patients were free of target vessel failure (defined as a composite of acute procedural failure, target vessel revascularization, myocardial infarction or death at six months). Only three MACE were observed within the first 30 days after stent implantation. A low left ventricular ejection fraction and the percentage stenosis pre-procedure were identified as statistically significant (p<0.05) predictive factors for MACE. The incidences of MACE and target vessel failure after six months follow-up were lower than that seen in other studies, and target lesion revascularization rates were low. PMID- 10825764 TI - Measurement of myocardial fractional flow reserve during coronary angioplasty in infarct-related and non-infarct related coronary artery lesions. AB - Myocardial fractional flow reserve (FFRmyo) has been demonstrated to be a useful method for determining the physiologic importance of a given coronary lesion. However, the reliability of the FFRmyo measurement is unknown in infarct-related arteries (IRA). The aim of this study was to measure and correlate the FFRmyo results of 14 consecutive patients who had recent acute myocardial infarction (AMI) (Group 1) with 14 consecutive patients who didnOt have AMI (Group 2) before and after percutaneous transluminal coronary angioplasty (PTCA). Quantitative coronary angiography (QCA) and FFRmyo measurements were determined both before and after optimal PTCA for all patients. FFRmyo was measured by use of a 0.014 inch guidewire as the ratio of the pressure distal to the target lesion to the aortic pressure taken during the maximal hyperemia induced by intracoronary adenosine. There were no differences between the two groups related to gender, target artery reference diameter, minimal luminal diameter and percent diameter stenosis of the vessel both before and after PTCA. While FFRmyo results after PTCA were not different between the groups, they were statistically different before PTCA (Group 1: 77.6+/-5.4%, Group 2: 63.3+/-8.4%; p<0.001). Although QCA determined percent diameter stenosis revealed a significant degree of stenosis (66.5+/-10.5%) for Group 1, FFRmyo values were higher than 75% (77.6+/-5.4%), indicating insignificant stenosis. Thus, it was concluded that FFRmyo measurements before PTCA were significantly different between IRA and non-IRA and that the method may not be valid for the determination of stenosis significance in IRA. PMID- 10825766 TI - Coronary artery bypass surgery on small patients N commentary PMID- 10825765 TI - Coronary artery bypass surgery on small patients. AB - Little research exists on the outcome of smaller patients who undergo coronary artery bypass graft surgery. The purpose of this study was to evaluate the outcomes of cardiac surgery in smaller patients (males with body surface area of 1.6 square meters or less, and women with 1.5 meters or less). A consecutive series of 4,358 patients undergoing bypass surgery was evaluated. Of these, 246 (5.6%) were classified as small patients. Smaller patients were more likely to be women, older and of Asian ethnicity. They were less likely to have undergone a prior cardiac surgery. Smaller patients were less likely to receive an arterial conduit (74% versus 99%; p<0.00001). Rates of post-surgery complications differed between small and normal size patients, with smaller patients more likely to require prolonged ventilator support (p <0.05), more likely to have acute renal failure (p<0.0001), more transfusions and re-operation for bleeding (p<0.05), higher death rate (5.7% versus 2.6%; p<0.01) and longer length of hospital stay (11.4 versus 8.3 days; p<0.00001). In multivariate analyses evaluating factors related to death, emergent surgery, poor ejection fraction and older patient age were independently related to mortality. Small body surface area was not an independent predictor. The results of this study indicate that smaller patients do have poorer outcomes associated with coronary artery bypass surgery. However, 90% of the smaller patients did have an event-free surgery. Surgeons may need to monitor these patients more closely and anticipate the increased risk and cost that is associated with this group. PMID- 10825767 TI - The Jomed Covered Stent Graft for coronary artery aneurysms and acute perforation: a successful device which needs careful deployment and may not reduce restenosis. AB - The Jomed Covered Stent Graft (Jomed International AB, Helsingborg, Sweden) is marketed for treatment of coronary artery aneurysms, perforations, dissection or thrombus. Three cases are presented, two with aneurysms, one with an acute coronary perforation. Intravascular ultrasound (IVUS) identified the need for high-pressure deployment of the stent. Although it has been suggested that this stent might lead to reduced rates of restenosis, one case later developed proliferative and occlusive in-stent restenosis and another suffered stent thrombosis at one month shortly after discontinuing clopidogrel. This niche stent clearly has an important role, but high-pressure deployment, IVUS evaluation and prolonged antiplatelet therapy are strongly recommended. PMID- 10825768 TI - Complications from Sinus of Valsalva aneurysm--a case of stable ischemia from compression of the left main stem and one of surgical recurrence. AB - Two cases of Sinus of Valsalva aneurysm with unusual complications are presented. Left main stem compression from an expanding Sinus of Valsalva aneurysm usually carries a poor prognosis and aggressive surgical management has been proposed. We describe a case with stable myocardial ischemia over four months from left main stem compression. Surgical repair of Sinus of Valsalva aneurysm is usually curative, but our second case demonstrates the pitfalls of inadequate surgical management. PMID- 10825770 TI - Stent grafting for coronary arteriovenous fistula with adjacent atherosclerotic plaque in a patient with myocardial infarction. AB - Coronary arteriovenous fistula is a relatively common congenital anomaly of coronary artery. Currently, percutaneous transcatheter embolization and surgical ligation are two commonly utilized management techniques. Stent grafts have recently been applied for various coronary lesions such as coronary perforation and coronary aneurysm. We report a case of coronary arteriovenous fistula with adjacent atherosclerotic plaque successfully treated with coronary stent grafting in a patient who presented with acute myocardial infarction. PMID- 10825769 TI - Acquired internal mammary artery to pulmonary artery fistula following bypass surgery. AB - We report two patients with acquired fistulas between the internal mammary and pulmonary arteries after coronary bypass surgery. This is a rare complication of bypass surgery and may be a cause of recurrent angina postoperatively due to coronary artery steal. A table of all the cases reported in the literature is compiled. PMID- 10825771 TI - Radiation safety of personnel during catheter-based Ir-192 coronary brachytherapy. AB - Catheter-based brachytherapy using Ir-192 seed sources has shown significant reduction in the rate of restenosis among patients with coronary in-stent restenosis. High-energy gamma rays from Ir-192 raise some radiation safety issues of personnel. The aim of this study was to fully analyze the radiation safety issues associated with Ir-192 brachytherapy in the cardiac cath lab environment. Measurements were made to assess the penetrating ability of Ir-192 gamma rays through tissues, concrete and lead. Radiation exposure levels were measured around a large number of patients undergoing Ir-192 brachytherapy. Personnel were carefully monitored for any additional dose received from brachytherapy for the last five years covering > 500 cases. Our results showed that with a proper radiation safety program in place, the dose to cath lab staff was negligible. It was concluded that radiation safety of personnel was easy to maintain during catheter-based coronary brachytherapy using Ir-192 seed sources. PMID- 10825772 TI - Studies of cardiac function and myocardial tissue characterization. AB - The heart can be studied using ultrasound techniques. The shape of the heart, its chambers, wall thicknesses, wall tissue characteristics as well as motion of walls and valve leaflets are all diagnostic information. In addition, the blood velocity and its timing within the cardiac cycle is an important diagnostic tool. In the present paper focus will be limited to the analysis of the left ventricular function as observed with two-dimensional and three-dimensional echocardiography and the characteristics of backscattered ultrasound information from the left ventricular chamber wall. Function of the heart is often studied by observation of local wall motion or comparison of chamber volume in maximum and minimum shapes during the cardiac cycle (ejection fraction). Integrated backscatter from the wall is described in examples of cardiac transplantation and hypertrophy. Study of cyclic variation of frequency-dependent attenuation and integrated backscatter indicates that these are independent parameters. PMID- 10825773 TI - Design of a surface replacement prosthesis for the proximal interphalangeal joint. AB - A surface replacement finger joint prosthesis was designed specifically for the proximal interphalangeal joint (PIPJ). The two-piece design consisted of a bi condylar proximal phalangeal head and a conforming bi-concave middle phalangeal base. The bearing surfaces were designed as close to the original anatomy of the PIPJs as possible, using detailed information obtained from a previous anatomical study of 83 PIPJs by the present authors. Four sizes of prosthesis were designed with maximum head diameters of 7, 8, 9 and 10 mm. Fixation of the joint prosthesis was achieved by an interference fit between the stems of semicircular cross-section and the phalangeal bone shafts. The main considerations for the stem designs were the offset from the centre of rotation, angle of inclination, length, and cross-sectional shape and size. It is proposed that the two components will be made from cross-linked polyethylene (XLPE) because it can be injection moulded to produce the complex shapes of the joint prosthesis. In addition, XLPE against itself has shown comparable wear rates with stainless steel against ultra-high molecular weight polyethylene from previous work by Joyce et al. PMID- 10825774 TI - Robotic-assisted internal fixation of hip fractures: a fluoroscopy-based intraoperative registration technique. AB - The internal fixation of proximal femoral (hip) fractures is the most frequently performed orthopaedic surgery procedure. When using a sliding compression hip screw, a commonly used fixation device, accurate positioning of the device within the femoral neck-head is achieved by initially drilling a pilot hole. A cannulated component of the hip screw is then inserted over the guide wire (surgical drill bit), which is used to drill the pilot hole. However, in practice, this fluoroscopically controlled drilling process is severely complicated by a depth perception problem and, as such, a surgeon can require several attempts to achieve a satisfactory guide wire placement. A prototype robotic-assisted orthopaedic surgery system has therefore been developed, with a view to achieving accurate right-first-time guide wire insertions. This paper describes the non-invasive digital X-ray photogrammetry-based registration technique which supports the proposed robotic-assisted drilling scenario. Results from preliminary laboratory (in vitro) trials employing this registration technique indicate that the cumulative error associated with the entire X-ray guided robotic system is within acceptable limits for the guide wire insertion process. PMID- 10825775 TI - Adsorption control of synovia constituents on artificial joint materials by means of an electric field: evaluation of tribological characteristics. AB - The supplemental lubrication mechanism by means of an electric field was proposed to reduce friction and wear for the advanced joint prosthesis with the low elastic modulus bearing surface. The possibility of application of this mechanism to the prosthesis was investigated by the fundamental and experimental procedures in simplified sliding conditions. Conductive silicon rubber was used as the low elastic modulus surface. The counterfaces were a titanium alloy and a stainless steel. Protein (gamma-globulin) in lubricant appeared to cause the tribological characteristic to deteriorate in the mixed lubrication regime. High friction seemed to be brought about by the obstruction against the entraining of the fluid flow and the high shear stress due to the microbonding between the asperities of bearing surfaces, which were derived from the adsorbed protein on the hydrophobic surfaces. The repulsive force between the adsorbed film and the bearing materials by means of the d.c. electric field, and the continuous change in polarity on the surface by means of the a.c. electric field appeared to affect the adsorbed film adjacent to the bearing material, so that friction and wear were varied. PMID- 10825776 TI - The development of a long, dual-platform triaxial walkway for the measurement of forces and temporal-spatial data in the clinical assessment of gait. AB - Force-plate measurement of the ground reaction force (GRF) has, for many years, been considered a vital component of the comprehensive assessment of human gait in the clinical context. For example, the data can be used in the adjustment of prostheses and orthoses and in identifying the mechanisms underlying a gait dysfunction. However, commercial force plates are usually only capable of measuring GRF data from one step in a single traverse. That can lead to problems of 'targeting' and, with less able subjects, fatigue before the necessary data have been collected. Previous work at the University of Surrey resulted in a prototype dual-platform force walkway capable of measuring the vertical component of the GRF and estimating the position of application of that force for multiple foot contacts in a single traverse. In addition, temporal-spatial information, e.g. speed and step length, could also be determined. This paper describes the development of a longer walkway that can measure the three orthogonal components of the GRF and provide and a more accurate estimate of the position of application of that force. Software to allow the rapid reduction of gait data to useful clinical information has also been developed. PMID- 10825777 TI - A study on dental vacuum system losses. AB - An experimental study was conducted to document system head losses for dental vacuum systems. Experiments were performed to estimate pressure losses and pressure loss coefficients for pertinent dental treatment room hoses and fittings in high-volume evacuation systems. Pressure losses for some polyvinyl chloride (PVC) elbows used in dental plumbing systems were also measured and presented. Friction head losses in straight PVC piping are calculated and documented in graphical form. Calculated values for friction losses are found to correlate exceptionally well with measured experimental data. PMID- 10825778 TI - Performance tests of vacuum pumps in small and large dental facilities. AB - A study was conducted to compare and assess the performance of various vacuum pumps in small and large dental clinic operations. A four-chair dental clinic operation was set up at the laboratory to simulate a small dental clinic environment. The performance of two dry and two wet pumps was assessed in the laboratory via extensive performance data acquisition tests. Data were also collected in two small dental clinics of practising dentists. Large dental system performance tests were conducted at the Ellsworth Air Force Base and Minot Air Force Base dental clinics. At each clinic, data were collected with both the existent turbine pump and a rotary vane RAMVAC pump. PMID- 10825779 TI - Gender differences in erotic plasticity: the female sex drive as socially flexible and responsive. AB - Responding to controversies about the balance between nature and culture in determining human sexuality, the author proposes that the female sex drive is more malleable than the male in response to sociocultural and situational factors. A large assortment of evidence supports 3 predictions based on the hypothesis of female erotic plasticity: (a) Individual women will exhibit more variation across time than men in sexual behavior, (b) female sexuality will exhibit larger effects than male in response to most specific sociocultural variables, and (c) sexual attitude-behavior consistency will be lower for women than men. Several possible explanations for female erotic plasticity are reviewed, including adaptation to superior male political and physical power, the centrality of female change (from no to yes) as a prerequisite for intercourse, and the idea that women have a milder sex drive than men. PMID- 10825780 TI - Gender differences in erotic plasticity--evolutionary or sociocultural forces? Comment on Baumeister (2000) AB - R. F. Baumeister (2000) argued that there are gender differences in erotic plasticity, meaning that women are more influenced by cultural and social factors than men are. He attributed the gender difference in erotic plasticity to evolutionary, biological forces. We propose an alternative account of the data using a multifactor sociocultural model that rests on 4 assertions: (a) Men have more power than women on many levels including the institutional and the interpersonal levels, (b) education increases women's power, (c) groups with less power (women) pay more attention to and adapt their behavior more to the group with more power (men) than the reverse, and (d) gender roles powerfully shape behavior, and heterosexuality is a more important element of the male role than the female role. PMID- 10825781 TI - Beyond artificial, sex-linked distinctions to conceptualize female sexuality: comment on Baumeister (2000) AB - The authors comment on three aspects of R. F. Baumeister's (2000) theoretical article on female sexuality. Questioning the predominance of nature versus cultural factors in accounting for sexual outcomes for men and women, the authors draw attention to the similarities (as opposed to differences) in the sexual attitudes, behaviors, and responses of men and women, and directly question the suggestion of "controlling" women's sexual attitudes, behaviors, responses, etc. to meet social needs for change. PMID- 10825782 TI - Nature, culture, and explanations for erotic plasticity: reply to Andersen, Cyranowski, and Aarestad (2000) and Hyde and Durik (2000) AB - R. F. Baumeister's (2000) article on erotic plasticity was criticized by B. L. Andersen, J. M. Cyranowski, and S. Aarestad (2000) for not being biological enough and by J. S. Hyde and A. M. Durik (2000) for being too biological. Both critiques were based on drawing a polarized caricature of R. F. Baumeister's actual view, although the two caricatures are opposites. Actually, neither commentary questioned the gender difference R. F. Baumeister documented; rather, the dispute is about how to explain it, which is indeed a challenge remaining for further work. Although both commentaries provided valuable suggestions about how to approach an explanation, neither approach can provide a coherent account until various theoretical problems are resolved and seemingly contrary empirical findings are addressed. PMID- 10825783 TI - Maxims or myths of beauty? A meta-analytic and theoretical review. AB - Common maxims about beauty suggest that attractiveness is not important in life. In contrast, both fitness-related evolutionary theory and socialization theory suggest that attractiveness influences development and interaction. In 11 meta analyses, the authors evaluate these contradictory claims, demonstrating that (a) raters agree about who is and is not attractive, both within and across cultures; (b) attractive children and adults are judged more positively than unattractive children and adults, even by those who know them; (c) attractive children and adults are treated more positively than unattractive children and adults, even by those who know them; and (d) attractive children and adults exhibit more positive behaviors and traits than unattractive children and adults. Results are used to evaluate social and fitness-related evolutionary theories and the veracity of maxims about beauty. PMID- 10825784 TI - A meta-analytic review of sex differences in facial expression processing and their development in infants, children, and adolescents. AB - Quantitative and qualitative reviews of the literature on sex differences in facial expression processing (FEP) have yielded conflicting findings regarding children. This study was designed to review quantitatively the literature on sex differences in FEP from infancy through adolescence and to evaluate consistency between the course of FEP development and predictions derived from preliminary theoretical models. Results, which indicate a female advantage at FEP, are consistent with predictions derived from an integrated neurobehavioral/social constructivist model. These findings suggest a need for research examining both neurological maturation and socialization as important factors in the development of sex differences in FEP and related skills. Possible directions for future study are discussed, with emphasis on the need to integrate the infant literature with research focused on older children and adults. PMID- 10825785 TI - Proposal of a four-dimensional model of social response. AB - Descriptive models of social response attempt to identify the conceptual dimensions necessary to define and distinguish various types of influence. Building on previous approaches, the authors propose a new response model and demonstrate that a minimum of 4 dimensions is necessary to adequately provide for such influence phenomena as conformity, minority influence, compliance, contagion, independence, and anticonformity in a single model. In addition, the proposed model suggests 5 potential types of response that have not been previously identified. These new types suggest directions for future research and theoretical development. Selected empirical evidence is reviewed in support of the validity and integrative power of the proposed model. PMID- 10825786 TI - [Determination of immediate hypersensitivity to pollen in allergic patients in the Federal District]. AB - AIMS: The immediate hypersensibility immediate mediated by IgE represents the damage mechanism type 1 according to the original classification of Gell & Coombs. MATERIAL AND METHOD: Observational study, descriptive, transverse in the one which were checked the files of 1250 patient of the Allergy Service of the Hospital Infantil de Mexico Federico Gomez with diagnostic of rhinitis allergic; asthma; asthma and rhinitis allergic. RESULTS: 1250 patient studied 468 were of the feminine sex (37.44%) and 782 (62.58%) of the masculine with a mean age in either case of 7.9 +/- 3.5 SD. With respect to result of cutaneous test 154 (12.32%) were negative and 1096 (87.68%) were positive to one or more antigens. CONCLUSIONS: The obtained results indicate the importance of accomplishing greater number of studies that relate not only the percentages of positive in the determination of immediate hypersensitive, but also to the frequency of pollens in the means to the one which is unfolded the patient, together with the stages of pollination throughout the Mexican Republic, in pediatric population as well as in adult, to effect cutaneous tests selects according to the zone and to stablish a specific treatment and adapted for each patient. PMID- 10825787 TI - [Changes in intraocular pressure with the use of topical nasal mometasone furoate]. AB - OBJECTIVE: The objective of the study was to valuation if the use of topical Mometasone Furoate for the treatment of rhinitis provokes an elevation of the intraocular pressure. MATERIAL AND METHOD: It was comparative, double blind, experimental, prospective and longitudinal. To measure the intraocular pressure of the patients at the third week, sixth week, three month, six month and one year. RESULTS: There were some modifications in the intraocular pressure, but without exceeding the normal parameters. CONCLUSIONS: Mometasone Furoate is safe and does not cause an increment in the intraocular pressure. PMID- 10825788 TI - [Frequency of association of rheumatoid factor in patients with breast cancer]. AB - AIMS: It has been reported that some patients with cancer present auto-immune phenomenon mediated by auto-antibodies, suggesting a relationship between auto immunity and cancer. Our interest was to determine the frequency of association of rheumatoid factor and breast cancer. MATERIAL AND METHOD: Fifty patients were studied, 3 on stage 111 and 19 on stage IV. Rheumatoid factor was measured in all of them. Auto-antibodies were measured by ELISA. The clinical files of all the patients were reviewed to determine the presence of metastases (osseous, pulmonary, CNS and hepatic) as well as the histological type of the cancer to correlate the expression of the rheumatoid factor with the patients' clinical status. RESULTS: Four (12.9%) out of the 31 patients on stage 111 had positive rheumatoid factor, while 9 (47.3%) out of 19 patients on stage IV had positive rheumatoid factor. The mean age of the patients on stage 111 with positive rheumatoid factor was 48 years, while the mean age of the patients on stage IV with positive rheumatoid factor was 53 years. Patients on stage 111 with positive rheumatoid factor only had local-regional metastases while patients on stage IV with positive rheumatoid factor had distant metastases. The predominant histological type was adenocarcinoma. CONCLUSION: In this study it is shown that breast cancer on the most advanced stages have higher expression of rheumatoid factor, and more clinical derangement with higher levels of rheumatoid factor expression. The proposal of auto-antibodies as predictors of the severity of the cancer requires further studies on several types of cancer. PMID- 10825789 TI - [Anamnesis and skin test to prevent fatal reactions to iodinated contrast media]. AB - AIMS: The phenomenon of allergic and toxic reaction to the means of yodates contrast occurs with greater frequency than what registers the literature. For this motive was effected the present study. MATERIAL AND METHOD: 178,439 cases of patients exposed to iodinated contrast media (ICM) corresponding to 137,147 to patients for excretory urography and 41,292 to cholangiography were studied. A direct interrogation making emphasis on personal antecedents of allergy to contrast media, iodinated products usage, nervous system and cardiovascular illnesses was made. Cutaneous test for iodinated contrast media was applied. If the result was positive no study with ICM was made or it was made with special guidelines. RESULTS: 4,302 positive and 1,276 false negatives were found, 287 positive cases were premedicated and preventive medication was administered in cases of urticaria asthma/and angioedema with negative cutaneous test histories. There were no deaths registered. CONCLUSIONS: The means of yodate contrast are susceptible of unfettering adverse reactions of pharmacological type or immunologic. For this reason is emphasised the usefulness of the anamnesis and the tests of intradermorreaction to detect cases of serious reaction. PMID- 10825790 TI - [Degree and duration of bronchodilatation with an agonist beta 2 administered alone versus an agonist beta 2 administered with ipratropium bromide in children with acute asthma]. AB - BACKGROUND: The asthma is the chronic illness of the most frequent respiratory tract of the pediatric age. MATERIAL AND METHOD: Prospective study, longitudinal, double blind, at random with 40 patient (split into two groups) with asthmatic crisis with ages of 8 to 15 years. To all was them accomplished tests of effected pulmonary function of basal way and subsequent to the inhalation of the medication. The medication were administered in two inhalations by three steps and with intervals of 10 minutes between each dose. It was considered that the treatment was effective if the improvement of the pulmonary function tests (VEF1) were presenting an increase of the 15% of their initial value. RESULTS: For each one of the tests accomplished in the different times were found meaningful increases in VEF1 (p < 0.05) in both groups compared with the basal and stayed meaningful until eight hours after the administration of both therapeutic plans. CONCLUSION: The bronchodilator effect of the salbutamol alone and in combination with bromide of ipratropio is similar in intensity and in action time demonstrated by the VEF1. PMID- 10825791 TI - [Treatment with montelukast in children with asthma. Test trial]. AB - OBJECTIVE: To study of antileukotrienes with participation in the genesis of the asthma appeared in the last years. MATERIAL AND METHOD: 17 children with a mean age of 10.29 years, male seven and ten girls. They were tried with montelukast 5 mg/day the 12-year-old smellers and with 10 mg/day the such age greats, during a period middle to 5.7 months. Ten of they had a concomitant treatment fluticasona, budesonide, salmeterol or formoterol. Was valued the clinic improvement, the saving in medication and the sound effects. RESULTS: a) Clinic improvement in 82.3%; b) elimination of medication associated in 90% and c) sound effects: in a case drowsiness and in other abdominal pain. CONCLUSIONS: Montelukast seems be a very useful product in the treatment of the bronchial asthma in children since generates a saving of corticoids and longtime beta 2 in the mild or moderate asthma, associate or not with the inmunoterapia. PMID- 10825792 TI - [New protocol for memory-related potential]. AB - A new P300 testing protocol for evaluating memory function is proposed. Four stimuli (S1, S2, S3 and S4) are presented to the subject at 1.5 sec intervals and the subject is instructed to judge whether S4 was the same (80%) or not (20%) as S1, as well as whether S3 was the same (80%) or not (20%) as S2. The S2-S3 comparison acts as an attention shifting mechanism, and by inserting this comparison between S1 and S4, the subject must memorized S1 at the memory site in the brain until S4 is presented. This protocol was performed using both auditory and visual stimulations with 24 college students (age range: 19-28 years old) and a patient (23 years old) with bilateral medial temporal lobe lesions caused by limbic encephalitis. The P300 brain potentials after S3 and S4 were compared. The Wechsler memory scale-revised (WMS-R) test was also assessed in all subjects. The latency of P300 after S4 had a significantly (p < 0.01) longer than that after S3. The latency of P300 after S4 in the patient, who showed the severe auditory as well as the mild visual memory impairment, was longer than the mean + 2 SD in the normal subjects for auditory stimuli, but not for visual stimuli, whereas the latency of P300 after S3 in the patient was longer than the mean + 2 SD in the normal subjects only for visual stimuli. The brain potential after S4 might be more reflective of a degree of memory disorder as compared with that after S3. Furthermore, the latency of P300 after S4 showed a more significant correlation with some WMS-R scores (verbal memory, general memory and verbal delay) than the latency of P300 after S3. These results suggest that measuring the P300 latency after attention shifting in the new protocol is useful for the evaluation of memory functions. PMID- 10825793 TI - [MRSA infection control in the wards for progressive muscular dystrophy: the effects of encouraged handwashing]. AB - There are currently 27 national hospitals with the wards for progressive muscular dystrophy (PMD) in Japan. Today, most patients in these wards are severely motor handicapped and complicated with respiratory and/or cardiac failure. Malnutrition, dysphagia, insufficient respiratory tract clearance are common problems that cause fragility to infection. Although progress in the treatment of infection have remarkably prolonged their life-span, frequent use of antibiotics is a major factor for occurrence of drug resistant bacteria. Since we had the first case of methicillin resistant Staphylococcus aureus (MRSA) infection in 1994 in our hospital, the number of MRSA carriers increased year by year. To prevent the spread of MRSA, we revised our infection precaution manual and surveyed its consequence. We encouraged handwashing of staffs, introduced green tea in cleansing genital region and abolished the use of a private room except a patient with severe seborrheic eczema. The revision of the manual resulted in decrease of MRSA carriers. The surveillance revealed that many patients had MRSA in genital regions, although there were no relationships between colonization of MRSA and severlity of disability or complications. It was strongly suggested that toilettary care by the hands of nursing staffs was a major factor of transferring MRSA. Our study revealed that encouragement of handwashing is more powerful in preventing the spreading MRSA, and more favorable for quality of lives of PMD patients than isolating the patients. PMID- 10825794 TI - [A case of diabetic polyneuropathy complicated with entrapment neuropathy of the bilateral ulnar nerves due to osteoarthrosis at the elbow]. AB - We report a 61-year-old man with diabetic polyneuropathy and bilateral ulnar nerve palsy due to osteoarthrosis in the elbow. He was diagnosed as having non insulin dependent diabetes mellitus (DM) at 40 years of age. At 56 years of age, he developed muscle atrophy and weakness predominantly in the distal parts of his upper limbs. A neurological examination showed him to have severe atrophy and weakness in the muscles innervated by the ulnar nerve bilaterally. He also had paresthesia on the distal parts of all four limbs. Superficial and deep sensory deficits were observed in the lower limbs. A motor nerve conduction study showed a marked reduction in the motor conduction velocity as well as in the amplitude of the action potentials of both ulnar nerves. Roentgenograms of the elbow joints and grooves for the ulnar nerve revealed marked osteophyte formation bilaterally. The bilateral ulnar nerve palsy was thus considered to be due to the entrapment of the nerve by the osteophyte. Since several studies have suggested the existence of a relationship between DM and osteoarthropathy, it is important to check for the possible presence of osteoarthrosis in cases of diabetic neuropathy complicated with entrapment neuropathy. PMID- 10825795 TI - [Complex visual hallucination in the quadrantanopsic field in a patient with a medial temporal hematoma following left putaminal hemorrhage]. AB - A 56-year-old man with a left medial temporal lobe hematoma from the left putaminal hemorrhage presented with complex visual hallucinations in the right quadrantanopsic field. In the right superior quadrantanopsic field, he saw colored and formed hallucinations of plants, bamboo thickets and a stature of bodhisattva. Most of these hallucinations were what he had actually seen before. The visual hallucinations disappeared when he gazed them with saccadic eye movements. EEG showed no epileptic discharges. MRI revealed hematoma from the lower putamen to the medial temporal lobe including amygdala on the left. 123I IMP SPECT showed hypoperfusion in the left temporal lobe and in the left occipital and parietal lobes. We speculate that the hallucinations of the present case were produced by the dysfunctions not only of the temporooccipital and temporoparietal regions but also of the posterior inferotemporal cortex which has strong interactions with amygdala and works as the visual memory center. PMID- 10825796 TI - [An unusual case of epilepsy exhibiting gelastic seizure, simple visual hallucination, and transient swelling of the left parieto-occipital region]. AB - We report a 74-year-old man with gelastic seizure, simple visual hallucination, and adversive seizure. The patient described his visual hallucinations as "rotating light like a firefly" and "mimicking a stream". Brain CT scan showed a transient swelling as well as low density of a left parieto-occipital region. Electroencephalographic study revealed spikes and fast waves beginning at left occipital region. Although temporal lobe and hypothalamic lesions (especially hypothalamic hamartomas) are well known as origins of gelastic seizures, we could not find any report that described a series of occurrence of gelastic seizure and simple visual hallucination. Usually, simple visual hallucination is thought to occur in occipital lesion. In our case, it is possible that gelastic seizure and simple visual hallucination are related to the epileptic discharge from occipital lesion directly or indirectly. The reversible brain swelling with low density seen in the present case might be caused by cytotoxic edema due to status epilepticus. PMID- 10825797 TI - [Chronic encapsulated intracerebral hematoma in thalamus with incongruous right homonymous hemianopia: a case report]. AB - We presented a patient with chronic encapsulated intracerebral hematoma. This 49 year-old woman suffered from visual disturbance, and slowly progressive right hemiparesis, sensory disturbance of the right extremities and incongruous right homonymous hemianopia over 2 months. Computed tomography scanning showed high density area and ring enhancement, and magnetic resonance imaging revealed mixed intensity on T1 and T2-weighted images in her left thalamus and internal capsule. Angiographic studies revealed no vascular anomaly or tumor stain. The pathologic pictures indicated well-encapsulated hematoma containing fresh and old hematomas in the left thalamus. Most reported cases of this disease had hematomas in the subcortex and no cases had similar visual disturbance. This report was prepared because this condition is uncommon and may remain unrecognized. PMID- 10825798 TI - [An autopsy case of malignant lymphoma who showed hearing loss and cavernous sinus syndrome in the first stage and hemophagocytic syndrome]. AB - A 48-year-old man was admitted to our hospital due to multiple cranial neuropathy such as vertigo, hearing loss of the left ear, visual loss of the left eye, left ptosis and total ophthalmoplegia of the left eye. Brain MRI revealed no abnormalities. After 10 days, he suddenly showed hemophagocytic syndrome with high fever, liver dysfunction, jaundice and pancytopenia. In spite of the treatment, he died of multiple organ failure after 26 days. At autopsy, malignant lymphoma cells of T cell origin directly invaded into the liver, spleen, intraperitoneal lymph nodes and other organs. Reports that direct invasion of lymphoma cells could be detected at the cranial nerves in the cavernous sinus are very rare. PMID- 10825799 TI - [A case of pericarotid syndrome with retention cyst in ethmoid sinus]. AB - A 42-year-old man noted pain on the left side of his forehead and left ptosis. On examination, he showed conjunctival hyperemia, ptosis and miosis in the left side, as well as hyperesthesia in the first branch of left trigeminal nerve. An MRI of his brain showed a retension cyst in the left ethmoid sinus. There was neither abnormalities in the parasellar lesion nor in the neck. We diagnosed him with pericarotid syndrome rather than cluster headache or Raeder syndrome. Five cases who had paranasal sinus lesions as a cause of cluster headache or Raeder syndrome have been reported. More cases are needed to clarify the association of retension cyst in ethmoid sinus and pericarotid syndrome. PMID- 10825800 TI - [Radiculopathy and reversible axonopathy in a tetraplegic patient with meningeal carcinomatosis]. AB - We report a 49-year-old tetraplegic woman with meningeal carcinomatosis secondary to breast cancer. Serial nerve conduction studies in the extremities revealed that the amplitudes of the sensory nerve action potentials (SNAP) and the compound muscle action potentials (CMAP) decreased rapidly within a few days after her admission. Plasma exchanges were done four times and restored the SNAP amplitudes to normal range. The CMAP amplitudes were also increased, but not to the normal range. These electrophysiologic changes were not associated with clinical improvement. Our patient's tetraplegia may be associated with a combination of two different mechanisms; reversible axonopathy caused by humoral factors that can be removed by plasma exchange, and irreversible radiculopathy due to direct cancer cell invasion. PMID- 10825801 TI - [A case of famotidine-induced aseptic meningitis]. AB - We reported a 50-year-old woman with a history of mixed connective tissue disease. She had two episodes of meningitis-like symptoms after taking famotidine and tiquizium bromide for treatment of gastric ulcer. From CSF findings (elevated pressure, increase of protein, polymorphonuclear pleocytosis, negative culture) and result of famotidine challenge test, we diagnosed her as a drug induced aseptic meningitis. Because she had taken tiquizium bromide several times previously without any side effects, we concluded that famotidine was a causative drug. She was recovered without sequelae within a few days following cessation of these drugs. This is the first report of H2-blocker induced aseptic meningitis. When we encounter a patient with aseptic meningitis who presents polymorphonuclear pleocytosis in CSF, we should suspect drug induced aseptic meningitis and take a history of drug medication including H2-blocker. PMID- 10825802 TI - [A case of codon 232 mutation-induced Creutzfeldt-Jakob disease visualized by the MRI-FLAIR images with atypical clinical symptoms]. AB - A 50-year-old man was admitted to our hospital because of disorientation and nocturnal restlessness. The patient presented chronically progressive dementia. No myoclonus or periodic synchronous discharge (PSD) was found over time, with abnormal evidence in MRI-FLAIR (fluid-attenuated inversion recovery) images alone. Brain biopsy and prion protein gene analysis led to the final diagnosis of Creutzfeldt-Jakob disease (CJD) induced by the point mutation at codon 232 (Met to Arg). To date the cases of M232R mutation-induced CJD have been reported to present clinical symptoms and pathological evidences similar to sporadic CJD cases, and differential diagnosis between the types has been believed to be difficult. Our case suggests that some types of CJD induced by point mutation at codon 232 cannot be easily inferred from clinical findings. PMID- 10825803 TI - [Pathogenesis of pulmonary thrombosis in Duchenne muscular dystrophy; a consideration from changes in serum CK and LDH levels]. AB - We assessed the hypercoagulative state in patients with Duchenne muscular dystrophy (DMD) by measuring peripheral blood creatine kinase (CK) and lactate dehydrogenase (LDH) levels. In 76 patients with DMD who had no complications during examination, the CK and LDH levels were examined in a total of 173 times. Both enzyme levels were elevated and highly correlated (R = 0.93). We examined the temporal alteration of serum CK and LDH levels during episodes of pulmonary thrombosis experienced in 7 DMD patients who fulfilled the following diagnostic criteria: 1. sudden onset of dyspnea, chest pain, and hypoxia; 2. abnormal coagulation profile or abnormal chest CT findings, such as peripheral wedged shaped shadow and; 3. pneumothorax ruled out from X-ray findings. Throughout the episode of pulmonary thrombosis, CK and LDH levels changed within the range of those from DMD patients who had no complications, however, the CK level was significantly elevated from the basal level (2.12 times, p = 0.0075) on the first day of the episode. The LDH level rose for several days, and subsequently both CK and LDH returned to the basal level. In patients with compartment syndrome, CK levels were reportedly elevated due to muscle fiber necrosis which lead a hypercoagulative state. Therefore, we propose that muscle fiber necrosis is one of causal factors in pulmonary thrombosis. In addition, elevated CK may be helpful in predicting pulmonary thrombosis. PMID- 10825804 TI - [Putaminal signal hypointensity on T2 weighted MR image in a patient with corticobasal degeneration]. AB - In the course of a 74-year-old woman with corticobasal degeneration (CBD), bilateral putaminal signal hypointensity appeared on T2 weighted MR image. This right-handed woman noticed progressive difficulty in writing and using chopsticks during two years. Neurological examination in February 1998 showed apraxia, rigidity, akinesia and myoclonus, which were more marked in the right hand side. MRI revealed atrophy of the left frontotemporal lobes. These findings were compatible to CBD. Her symptoms progressed slowly. On re-admission due to aspiration pneumonia in December 1998, she showed difficulty in swallowing and walking in addition to the signs and symptoms described above. MRI revealed bilateral putaminal signal hypointensity on T2 weighted image. In the literature, totally 9 cases in 2 reports showed similar putaminal signal hypointensity. Although this finding is unusual in CBD, advanced degenerative process in the putamen might have induced this change due possibly to the accumulation of iron secondary to the nerve cell loss. PMID- 10825805 TI - [Atypical Miller Fisher syndrome associated with antiphospholipid antibodies]. PMID- 10825807 TI - [Development of highly stereoselective reactions utilizing heteroatoms--new approach to the stereoselective Horner-Wadsworth-Emmons reaction]. AB - This article reviews a new approach to stereoselective Horner-Wadsworth-Emmons (HWE) reactions. The HWE reaction is one of the most efficient methods for the preparation of alpha,beta-unsaturated esters, which play an important role in the synthesis of biologically active compounds. The reactions of aldehydes with phosphonates bearing alpha-substituents which stabilize the carbanion, preferentially furnish the corresponding E-alkenes. However, the stereoselectivity of the HWE reactions with ketones has never been investigated in detail because of their low reactivity and low stereoselectivity. The conventional HWE reactions of aryl alkyl ketones with ethyl diethylphosphonoacetate in the presence of sodium hydride gave the corresponding alpha,beta-unsaturated esters with modest E-selectivity. On the other hand, the treatment of aryl alkyl ketones with ethyl diethylphosphonoacetate in the presence of Sn(OSO2CF3)2 and N-ethylpiperidine afforded alpha,beta-unsaturated esters in a highly Z-selective fashion. A significant improvement in the selectivity and yield was found when the Still's reagent, methyl bis(trifluoroethyl)phosphonoacetate, was used under Sn(II)-mediated conditions. On the basis of the experimental results, the high Z-selectivity in the Sn(II) mediated HWE reactions of aryl alkyl ketones with these phosphonates can be rationalized in terms of six-membered transition state involving Sn(II) chelation. Similarly, the HWE reactions of aryl alkyl ketones with ethyl 2-fluoro 2-diethylphosphonoacetate in the presence of Sn(OSO2CF3)2 and N-ethylpiperidine gave the corresponding alpha-fluoro-alpha,beta-unsaturated esters in a highly E selective manner. Finally, the Sn(II)-mediated asymmetric HWE reactions of isopropyl 2-fluoro-2-diethylphosphonoacetate with 4-tert-butylcyclohexanone in the presence of a chiral diamine is described. PMID- 10825806 TI - [Structure and biological activities of fungal beta-1,3-glucans]. AB - This paper summarizes the structure, biological activities, signaling, and metabolic degradation of fungal beta-1,3-glucans. Fungal beta-glucans exist both soluble and particulate forms. Conformation of the soluble beta-glucan was classified into three groups: triple helix, single helix and random coil. Fungal beta-1,3-glucans exhibit a variety of biological and immuno-pharmacological activities, and the significance of these activities is dependent on the structure, such as solubility in water, molecular weight, degree of branching, and conformation. Many of the activities, such as nitrogen oxide synthesis of macrophage and limulus factor G activation, were dependent on the conformation, but some of the others were independent, such as interferon-gamma and colony stimulating factor syntheses. These activities are beneficial and pharmacologically useful, while some strongly related to allergic and inflammatory adverse reactions. It should be noted that the beta-glucans, once administered into the body, remain mainly in the liver and spleen for a very long period of time. The activity, at least in part, is maintained during these periods. The biochemical mechanisms of the beta-glucan, especially in soluble form, mediating biological activity was still not clearly demonstrated. PMID- 10825808 TI - [Kinetic analysis of drug disposition and biological response]. AB - This review deals with pharmacokinetic/pharmacodynamic analysis of drugs. For the analysis of antipyretics, it was assumed that: (1) The rat body is divided into two compartments, core and skin. (2) Metabolic heat (M) is generated in the core compartment. (3) Heat loss by vaporization (V) is mainly originated from a respiratory effect and occurs in the core compartment. (4) At the skin compartment, heat is gained from the core compartment by conduction (K) and is transferred to the ambient air by radiation and convection. (5) Central nervous system commands efferent signals for M, K and V to change their values according to changes in afferent signals from core and skin temperatures. (6) The effect of antipyretics is shown as afferent signals to the controller. For loop diuretics, it was assumed that: (1) The diuretic rate can be correlated with the urinary excretion rate of diuretics. (2) If there is no intervention in a body fluid regulation system, the relationship between the diuretic rate and the corresponding urinary excretion rate can be described by a Hill equation. (3) Intensity of the body fluid regulation is also described by the Hill equation, in which the intensity is correlated with cumulative amount of drugs excreted in the urine. For neuromuscular blockade, assumptions were: (1) There exists an acetylcholine (ACh) compartment at a motor nerve terminal. (2) ACh in the compartment is eliminated by a first-order rate process. (3) All of the ACh in the compartment is released by one electrical stimulus. (4) The compartment is replenished by two kinds of ACh mobilization. One is a slow mobilization with a constant rate and the other is a momentary mobilization which takes place just after the release of ACh. (5) The released ACh is metabolized immediately after binding to receptors and causing a twitch response. For centrally acting drugs, the quantitative electroencephalographic (EEG) method was used as a surrogate measure of a pharmacological response. Signals from two electrodes fitted on the skull of rats were continuously measured, recorded and subjected to off-line analysis. Total amplitude from aperiodic analysis was taken as an EEG parameter. PMID- 10825809 TI - [Evaluation of time course and inter-relationship of inflammatory mediators in experimental inflammatory reaction]. AB - Inflammatory signs, such as heat, redness, swelling and pain, have been described from the Greek era. In these phenomena various endogenous active substances, i.e., inflammatory mediators, could cause and manifest vascular dilatation, a vascular permeability increase and sensitization of pain receptors, etc. In order to evaluate the roles of inflammatory mediators, we have studied the time courses of inflammatory reaction along with detection of various active substances directly or indirectly in the experimental animal model of pleurisy, such as rat carrageenin-induced, and zymosan-induced pleurisy. These pleurisies showed almost similar time courses of pleural exudate accumulation and neutrophil migration. However, mediators detected in the exudates of such pleurisies were different; in carrageenin-induced pleurisy bradykinin and prostacyclin (PGI2) caused exudate formation, while zymosan-induced pleurisy showed early degradation of mast cells and activation of complements, followed by an increase in platelet activating factor (PAF). In both pleurisies TNF alpha, IL-1, IL-6 and CINC (cytokine-induced neutrophil chemoattractant) appeared similarly in the exudates to cause chemoattractant for neutrophils. TNF alpha and IL-1 could stimulate to produce IL 6 and IL-8. While prostaglandins may regulate cytokine production via a cellular cAMP-dependent mechanism. Thus one should consider the time for application of anti-inflammatory drugs, such as cyclooxygenase inhibitor, indomethacin, since it causes increases in TNF alpha and IL-1 production by reducing PGI2 and prostaglandin E2 (PGE2) levels. In conclusion, inflammatory reaction has its own automatic regulation mechanism through complex cross talks between inflammatory mediators. PMID- 10825810 TI - [Toxicity of cadmium and lead on vascular cells that regulate fibrinolysis]. AB - Cadmium and lead are heavy metals that cause vascular lesions such as atherosclerosis and hypertension. Toxicity of cadmium and lead on the regulation of fibrinolysis by vascular-composing cells was investigated using a cell culture system. It was found that cadmium promotes the synthesis of plasminogen activator inhibitor-1 (PAI-1) whereas lead inhibits the synthesis of tissue plasminogen activator (t-PA) in vascular endothelial cells; consequently, both heavy metals reduced fibrinolytic activity in the liquid phase. The responses of endothelial cells to cadmium and lead were different from those to other heavy metals and the release of the fibrinolytic proteins from vascular smooth muscle cells and fibroblasts was perturbed by cadmium and lead in different manners. In conclusion, the present study showed that cadmium and lead exhibit their toxicity on fibrinolysis regulated by vascular cells in different manners among cell types and the individual cell types respond to cadmium and lead in different manners with respect to the release of fibrinolytic proteins. PMID- 10825811 TI - [Antihypertensive substance in the leaves of kumis kucing (Orthosiphon aristatus) in Java Island]. AB - The water decoction of leaves of kumis kucing, Orthosiphon aristatus (BL.) MIQ (Lamiaceae) which has been prescribed in Javanese traditional medicine (jamu) for the treatment of hypertension etc., was partitioned into a mixture of chloroform and water. The chloroform-soluble portion showed an inhibitory effect on the contractile responses on rat thoracic aorta smooth muscle stimulated with KCl beforehand, while the water-soluble portion showed no effect. The chloroform soluble portion was separated to afford a new benzochromene [orthochromene A (7)], two new isopimarane-type diterpenes [orthosiphonone A (10), orthosiphonone B (11)], and two novel migrated pimarane-type diterpenes [neoorthosiphol A (12), neoorthosiphol B (13)], together with eight known compounds (1-6, 8, 9). Among those thirteen substances, it was found that a major constituent in the water decoction of leaves, methylripariochromene A (5), exhibited a continuous decrease in systolic blood pressure after subcutaneous administration in conscious stroke prone spontaneously hypertensive rats (SHRSP). PMID- 10825812 TI - [Apoptotic cell death induced by actinomycin D arisen at different levels and cell cycle stages according to human cultured cell species]. AB - Nine kinds of human cultured cells, including fetus cells (smooth chorion trophoblast cells, amnion epithelial cells and HE-21), adult non-carcinoma cells (HCF), and carcinoma cells (KATO-III, COLO 201, Lu-134-AH, SK-OV-3 and SKG-3a) were stimulated with Actinomycin (Act.) D for 24 h. Apoptosis induction was investigated by agarose gel electrophoresis for DNA fragmentation analysis and by flow cytometric analysis of stained cells using in situ terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labeling TUNEL) staining techniques for the quantification of apoptosis, and simultaneously using propidium iodide for the gain of some information about cell cycle. By agarose gel electrophoresis, DNA fragmentation of these cells except amnion epithelial and SKG-3a cells was detected, depending on concentration of Act. D. Using flow cytometric analysis, these cells were separated into four groups according to the information about cell cycle. Group 1 included amnion epithelial and SKG-3a cells, which were TUNEL negative. In group 2, all cell populations at G0/G1 and G2/M phases of HCC, KATO III and SK-OV-3 were TUNEL staining positive. A portion of each G0/G1 or G2/M phase cell of Lu-134-AH and COLO 201 in group 3 was TUNEL stain positive. In group 4, G2/M phase cells of smooth chorion trophoblast cells and HE-21 were mostly stained and a small population of G0/G1 phase cells were also TUNEL stain positive. These results show that the stages of the cell cycle at which apoptosis was arisen by Act. D stimulation were significantly different depending on the cells types. PMID- 10825813 TI - Influences of 4-meta/MMA-TBB adhesive resin on osteodentinogenesis of transplanted rabbit dental pulp in vivo: immunohistochemical and electronmicroscopic studies. AB - The purpose of this study was to investigate the influence of 4-META/MMA-TBB adhesive resin (4-META resin) on osteodentinogenesis of transplanted pulp in vivo. Dental pulp was obtained from the incisors of adult rabbits. 4-META resin was applied to the pulp tissue, and the pulp tissue with 4-META resin was autotransplanted beneath the renal capsule with the pulp side touching the kidney. Pulp tissue alone was also transplanted as a control. The animals were sacrificed at 3, 7, and 14 days after the experiment, and the specimens were examined morphologically. At 3 days, proliferation of mesenchymal cells was observed, and alkaline phosphatase activity (ALP) and osteocalcin were detected throughout the entire transplanted pulp area. In the experimental case, a thin, highly electron dense zone and a granular layer were observed. Under this layer, only a cell-membrane-like structure, a cell with an unclear nucleus, a nucleus alone, and an organelle-like structure could be seen. Furthermore, an exudative layer with many neutrophils was observed, and apoptotic-body-like structures were also found in some areas. On days 7 and 14 in the control group, osteoblast-like cells had proliferated, and osteodentin formation was initiated throughout the entire transplanted pulp area. In the experimental cases at 7 and 14 days, the entire transplanted area had become osteodentin except for a thin fibrous layer under the 4-META resin. These results suggested that the components of 4-META resin such as MMA and 4-MET (A), which guide the polymerization, might cause degeneration of, but not disturb, the wound healing of the pulp tissue. PMID- 10825814 TI - In vivo and electron microscopic observations of the responses of the hepatic sinusoid to interleukin-1. AB - Interleukin-1 (IL-1), which is produced by Kupffer cells and sinusoidal endothelial cells, may play an important role in immunological and microvascular responses to a variety of stimuli in the liver. The responses of the hepatic microvasculature including phagocytic activity of sinusoidal lining cells to IL-1 alpha were examined in C57Bl/6 mice in vivo and using electron microscopy. One hour after recombinant mouse IL-1 alpha was injected at doses of 80 U, the low dose group, and 800 U, the high dose group, the phagocytic activity of sinusoidal lining cells showed significant differences between the two treated groups and between the two groups and the controls. In the low dose group, the numbers of sinusoids containing blood flow and of leukocytes adhering to the sinusoidal lining remained unchanged, but the former decreased and the latter increased significantly in the high dose group. Ultrastructurally, Kupffer cells that phagocytosed latex particles appeared to have decreased in number while the sinusoidal endothelial cells became phagocytic. A considerable number of leukocytes were seen adhering to the sinusoidal endothelium. These findings demonstrate that IL-1 alpha not only elicited sticking and plugging by leukocytes in sinusoids but also activated phagocytic functions in the hepatic sinusoidal endothelial cells. These endothelial responses are similar to those seen following FV3 virus infection, chronic administration of ethanol, or a combination of cocaine and ethanol, or during extracorporal perfusion, suggesting that IL-1 may participate in these responses. PMID- 10825815 TI - Clinical study of the pumping pressure changes in the temporomandibular joint space before and after arthroscopic surgery. AB - When arthroscopy of the temporomandibular joint (TMJ) is performed, we often observe differences in the resistance of hydraulic pressure during intra articular pumping. It can be speculated that intra-articular adhesion and contracture of the capsule and ligament may cause these differences in resistance. In order to measure the changes in intra-articular hydraulic pressure before and after arthroscopic ablation surgery, seven cases were examined by balloon pumping technique. We concluded that the values of intra-articular hydraulic pressure are implicated in the severity of the internal derangement of TMJ. PMID- 10825816 TI - Relationship between aging and recovery of the underlying mucosa immediately after removal of pressure: the first molar region in the mandible. AB - To investigate changes in the characteristics of edentulous mucosa in the alveolar ridge with aging, we applied pressure to the mucosa of the alveolar ridge in subjects of various ages and evaluated the recovery amount and time after the pressure was removed. Twenty-three subjects ranging from 23 to 79 years of age, who had lost the first mandibular molar tooth, were selected. Displacing pressure was applied to the mucosa corresponding to the mandibular first molar region. After release the pressure, we measured the recovery amount of mucosa from its displacement, which was expressed as the movement of a target which had been placed on the mucosa as measured by an indirect displacement measurement device. The levels of displacement were set at 0.1 mm, 0.2 mm and 0.3 mm, and the time to achieve the selected displacement level was set at 1 second. The initial recovery of the mucosa immediately after the pressure has been released was designated as the initial recovery amount, and the time needed for recovering to 90% of the previous state was designated as the final recovery time. Then we investigated the relationships between the measures of the initial recovery amount, the final recovery time, and the subjects' ages. The initial recovery amount of mucosa was lower and the final recovery time was longer in older subjects, and this tendency was exaggerated when the displacement was increased. PMID- 10825817 TI - Buccal mucosa ridging and tongue indentation: incidence and associated factors. AB - Buccal mucosa ridging and tongue indentation have been considered as one of the visible and reliable signs of bruxism. However, there have not been any reports justifying this relationship scientifically. Moreover, there have not been any studies reporting specific procedures to assess them. Thus, the purpose of the present study was to determine the clinical incidence of buccal mucosa ridging and tongue indentation and assess the possible relationship between certain factors that can influence their occurrence. A total of 244 (178 males and 66 females) dentulous adults from 20 to 59 years of age, who were employees at the Bank of Yokohama, were randomly selected. At first, the buccal mucosa ridging and tongue indentation were classified into three groups based in their intensity: none, mild, and severe. The incidence of both conditions in the different age groups, as well as the incidence by gender was evaluated. Furthermore, the possible relationships between buccal mucosa ridging and tongue indentation and age, gender, clenching awareness, grinding awareness, headache, neck stiffness, vertical dimension, temporomandibular joint (TMJ) pain to palpation, masticatory muscle tenderness to palpation, and the presence of premature contacts were evaluated using the chi-square test. A positive relationship was found between the occurrence of buccal mucosa ridging and tongue indentation and gender (p < 0.01); both conditions were observed more frequently in females than in males. A positive relationship was also found to age; the group between 20-29 years old showed the highest incidence. The vertical dimension had a positive relationship with the occurrence of both buccal mucosa ridging and tongue indentation. Other factors evaluated did not show any correlation. PMID- 10825818 TI - An analysis of management of private dental clinics in Japan from the survey of the current economy of medicine and dentistry. AB - We analyzed the current state of management of private dental offices in Japan using the 11th Survey of the Current Economy of Medicine and Dentistry conducted in September of 1997. The analysis was mainly performed by comparing the survey in 1997 with the survey conducted in 1989. The results show the medium-term tendencies in dental management. We also compared the data from dental offices with those from private medical offices without facilities for inpatients. The following are the results: the average income from dental practice for a dental office in 1997 was 4,050,000 yen, a decrease of 4.9% (about 210,000 yen) compared with the average income in 1989. The average income from treatment covered by insurance in 1997, which accounted for most of the total income, was 3,540,000 yen, a decrease of 2.8% compared with that in 1989. The average income from non insured treatment was 490,000 yen, which was also a decrease of 17.4%. The average income from medical practice for a medical office in 1997 was 1,970,000 yen more than the average income from dental practice of the same year. The average medical practice expenses and the average balance in medicine were higher than those in dentistry by 1,360,000 yen and by 600,000 yen respectively. Although the average income from medical practice slightly increased, the average wage costs increased by as much as 42.8%, resulting in a decrease of 21.2% in the average balance. Dental offices attempted to weather the decreased income from dental practice by raising the average fee paid by an insured patient. The analysis in the present study suggested that a decreasing number of patients and increasing dental practice expenses diminished the balance. PMID- 10825819 TI - VIP inhibits high voltage-gated calcium channel currents of hamster submandibular ganglion neurons. AB - In this study, we investigated the modulation of calcium channels by vasoactive intestinal polypeptide (VIP) in hamster submandibular ganglion (SMG) cells using the whole-cell patch clamp technique. VIP (1 microM) inhibited the high voltage gated calcium channels in the SMG neurons via activation of VIP receptors and G protein coupled to these receptors, but did not affect the low voltage-gated calcium channels. VIP at 1 microM reduced the peak amplitude of the maximum high voltage-gated calcium current by 26.0 +/- 6.2% (mean +/- S.E., n = 5) and slowed the rising phase of the calcium current. The time to peak of the maximum high voltage-gated calcium current was prolonged from 16.8 msec to 22.4 msec. In a representative cell, 1 microM VIP reduced the peak amplitude of maximum high voltage-gated calcium current by 50.5% and the amplitude at the end of voltage step after 300 msec by 51.9%. The VIP-induced inhibition of the calcium current was produced in a voltage-dependent manner. Inhibition was maximal (50.5%) at the level of -20 mV. Therefore, VIP inhibits both transient and sustained types of high voltage-gated calcium channels in the hamster SMG neuron. PMID- 10825821 TI - An undistinguished career in the Royal Army Dental Corps, 1943-947. PMID- 10825820 TI - Dental surgeons in the early days of anaesthesia. PMID- 10825822 TI - Milestones in dental education in the U.K. PMID- 10825823 TI - The development of dental schools--an evolutionary hypothesis. PMID- 10825824 TI - From naturalist to scientist: the world of F H Balkwill 1837-1921, the pioneer of dental articulation. PMID- 10825825 TI - Pain and new cultural diseases. AB - The purpose of this article is to recount a piece of medical history, even though it has been told many times before. Everyone has heard about patients who visit their doctors time and again with physical complaints without the physician being able to find an adequate organic cause. The oldest descriptions of this illness can be found on ancient Egyptian papyrus rolls, dating back to about the year 1900 BC (Shorter F. A history of psychosomatic illness in the modern era. New York: Macmillan; 1992). The need to tell these stories repeatedly can be attributed to the chameleonic nature of the illness which is difficult to spot because of its changing expression, which one could say takes on the appearance of its surroundings. Throughout history, humans have had the capacity to develop symptoms almost identical with the real diseases of the time. The term somatize is often used descriptively about patients with physical complaints without any recognizable organic basis. Shorter proposes that mankind has a shared "symptom pool" containing all existing symptoms. Patients who convert their mental state into bodily symptoms--somatization--take their symptoms from this symptom-pool, albeit it is possible for new symptoms to emerge. For centuries patients have presented with sensory symptoms such as paresthesias. In the 19th century patients began to exhibit motorial symptoms with attacks of hysteria. According to Shorte's theory, this multitude of manifestations should be perceived as culturally determined or as different expressions of the same underlying phenomenon. PMID- 10825826 TI - Scanning electron microscopy (SEM) investigation of selected desensitizing agents in the dentine disc model. AB - Dentine sensitivity is a painful clinical conditions that can affect up to 35% of the population at any one time. Both professionally available (in-office) or commercially available (over-the-counter) products have been used to treat dentine sensitivity. The aim of the present study was to investigate whether selected in-office desensitizing agents occluded dentine tubules in the dentine disc model. Both surface effects and tubule penetration of the five selected test products were examined by scanning electron microscopy. The results of the present study appeared to demonstrate that all of the applied desensitizing agents produced some occlusion of the tubules although the level of coverage and occlusion varied between the products. Of all the agents tested, ferric oxalate, the active ingredient of Sensodyne Sealant, produced crystal-like structures which occluded a higher proportion of the tubules across the dentine disc surface. ALL-BOND 2 and One-Step (both light-cured primer systems) produced similar crystal-like structures and, although coverage was not uniform across the disc surface, there was some reduction in tubule diameter. These three products, however, appeared to be more effective than either Butler Protect (potassium oxalate) or Oxa-gel (potassium oxalate in a gel) where there was a marked decrease in both the level of coverage and tubule occlusion. Both quantitative and functional studies are required in order to determine the effects of these agents on dentine permeability (fluid flow) as well as clinical studies to determine their effectiveness over time in reducing pain arising from dentine sensitivity. PMID- 10825827 TI - The antimicrobial effect of calcium hydroxide in root canals pretreated with 5% iodine potassium iodide. AB - Calcium hydroxide (CH) is often used as a routine interappointment dressing during the endodontic treatment of teeth with apical periodontitis. However, it fails to consistently produce sterile root canals. The present study was set up to find out whether an antimicrobial strategy including the use of CH could be made more effective if: 1) canals were pretreated with 5% iodine potassium iodide (IPI), and 2) the dressing period was extended up to 2 months. Fifty human teeth, with radiographically verified apical periodontitis, were microbiologically sampled. After chemomechanical preparation the canals were pretreated with IPI for 3-7 days. Teeth where microorganisms persisted were then treated with CH for 2 months. Following instrumentation and dressing with IPI, 43 bacterial strains were recovered in 22 of the teeth. Samples obtained after the CH dressing period disclosed growth of 13 facultative and two strict anaerobic strains in 10 teeth. Enterococcus faecalis was identified in two specimens. In conclusion, the present study gave no evidence for an increased antimicrobial effect of CH if it was left for longer periods in the root canal. Although pretreatment with IPI from a quantitative point of view did not seem to add antimicrobial power, it might reduce the frequency of persisting strains of E. faecalis. PMID- 10825828 TI - Endodontic treatment and prevalence of apical periodontitis in an adult population of Vilnius, Lithuania. AB - Apical periodontitis is prevalent in many Western populations and is frequently detected in connection with inferior quality of endodontic treatment. However, information about endodontic conditions from eastern European countries is limited. The aim of the present investigation was to study the prevalence of apical periodontitis and the quality of endodontic treatment in 35-44-year-old Lithuanians. The prevalence of apical periodontitis was 70%. An overall success rate of 65% was found for root-filled teeth while 56% of pulp amputations (pulpotomies) were considered successful based on a radiographic evaluation. The quality of the root fillings showed little impact on the treatment results, except for over-extended fillings, which were associated with a high number of failures. PMID- 10825829 TI - Muco-gingival and periodontal health recovery following reimplantation of teeth. AB - Reimplantation of avulsed permanent teeth following traumatic injuries is a widely accepted therapeutic measure; however, though periodontal healing is considered to be the major determinant of long-term survival of reimplanted teeth, the mucogingival reaction to reimplantation has never been assessed. In the present study, hygiene level and gingival inflammation of 13 reimplanted teeth were assessed according to the plaque index (PI) and the gingival index (GI) around both reimplanted and control teeth. The periodontal condition was evaluated by measuring probing depth and probing attachment level at the same location. The clinical parameters of the reimplanted teeth were compared with those of contralateral teeth with the help of the non-parametric Wilcoxon signed ranks test. There were no statistically significant differences in PI and GI scores at any points of measurements around both reimplanted and control teeth; neither did probing depths and probing attachment levels differ between reimplanted and contralateral teeth. The results of the present study indicated that the muco-gingival reaction following traumatic avulsion did not affect the long-term prognosis of the reimplanted teeth. PMID- 10825830 TI - Re-innervation in the canine periodontal ligament of replanted teeth using an antibody to protein gene product 9.5: an immunohistochemical study. AB - The re-innervation process in the periodontal ligament of replanted canine teeth was examined by immunohistochemistry for protein gene product 9.5 (PGP 9.5), a general marker for neurons, and by electron microscopy. Within 1 week of replantation, the periodontal fibers had regenerated, filling the narrow spaces between the alveolar bone and the root surface around the cervical and apical regions. Near the root apex, however, no PGP 9.5-immunopositive nerve fibers were found in the regenerated periodontal ligament except for those in the alveolar half of the ligament. At 2 weeks after replantation, many nerve fibers positive for PGP 9.5 had ascended the periodontal ligament from the thick nerve bundles located near the root apex. Fine nerve endings showing complicated ramification were also present in the apical region. By 3 or 4 weeks after replantation, the vascular network was regenerated and principal periodontal fibers were re established throughout the entire length of the periodontal ligament. The extensively ramified PGP 9.5-immunopositive structures had increased in thickness and density and showed characteristic tree-like profiles by 3 weeks. Electron microscopy confirmed that most of these structures were Ruffini-like endings, and demonstrated that such nerve terminals were almost regenerated by 4 weeks post replantation. These results indicated that, in the periodontal ligament of replanted canine teeth, the regeneration of the nerve fibers including mechanosensory receptors first showed signs of regeneration by 2 weeks following tooth replantation and proceeded rapidly thereafter. Regeneration of the periodontal ligament including fiber architecture as well as vascular and neural elements was almost complete by 4 weeks after replantation. PMID- 10825831 TI - Incidence of dentofacial injuries in children: a 2-year longitudinal study. AB - In this prospective study the incidence of dentofacial injuries in 199 children aged 8 to 10 years was analyzed. Dentofacial injuries were recorded by a questionnaire and a clinical examination. After the first examination, each subject was re-examined at yearly intervals over a period of 2 years. Incidence rates were calculated in percentages and the chi-square was applied to test statistical differences. The 95% probability level was used. Results showed that the overall incidence of dentofacial injuries was 45.2%. The percentages for boys and girls were 28.6 and 16.6 respectively. The chi-square test showed significant gender differences, while differences by age were not significant. The highest incidence was recorded for dental injury followed by facial injury, historical evidence of injury and dental and facial injury. PMID- 10825832 TI - Apical microleakage after lateral condensation, hybrid gutta-percha condensation and Soft-Core obturation: an in vitro evaluation. AB - The purpose of the present study was to evaluate the apical seal of root canals filled with a cold lateral condensation, a hybrid gutta-percha condensation technique and with Soft-Core obturators. Three groups of 35 maxillary anterior teeth and four control anterior teeth were treated with a crown-down/step-back technique. All roots were submerged in India ink for 1 week. The roots were cleared and the degree of linear dye penetration was measured. The mean apical leakage for the Soft-Core technique was at least twice as extensive as for the two other gutta-percha obturation techniques. No statistically significant differences were found between the two other condensation techniques. PMID- 10825833 TI - Delayed apical healing after apexification treatment of non-vital immature tooth: a case report. AB - We report the endodontic treatment of a non-vital permanent immature tooth in which unexpected complications such as exacerbation of apical periodontitis followed by external root resorption occurred after that the initial stages of the healing process were clinically and radiographically evident. After continued treatment stable repair was obtained. PMID- 10825834 TI - Failure of inferior alveolar nerve block in endodontics. AB - Analgesia is essential for successful completion of modern dental procedures. Standard inferior alveolar nerve block (IANB) is the primary method used to achieve mandibular analgesia. Difficulty experienced in obtaining satisfactory analgesia after IANB, especially of an acutely inflamed mandibular molar, remains a common clinical problem. Even when a proper technique is employed, clinical studies show that IANB fails in approximately 30% to 45% of cases. The reasons for failure are not fully understood. Anatomical considerations and abnormal physiological responses in the presence of inflammation as explanations for IANB failure are discussed in this paper. PMID- 10825835 TI - Influence of several factors on the success or failure of removal of fractured instruments from the root canal. AB - The influence of several factors on the success rate of removal procedures of fractured endodontic instruments was evaluated postoperatively. In 105 teeth with 113 fragments removal attempts were undertaken using a wide range of techniques and instruments. All cases were analyzed with special regard to the following factors: type of tooth and root canal, site of fragment in relation to root canal curvature, length of fragment, and type of fractured instrument. Success of treatment was defined as removal or complete bypassing of the fragment. Of 82 instruments in molars (maxillary: 32, mandibular: 50), 56 were removed or bypassed (max.: 26, mand.: 30). Of 16 fragments in premolars (max.: 12, mand.: 4), 8 could be removed or bypassed (max.: 6, mand.: 2). Of 14 fragments in canines and incisors (max.: 7, mand.: 7), 13 could be removed completely (max.: 6, mand.: 7). When the fragment was localized before the curvature 2 of 18 cases failed, when localized inside the curvature 13 of 31 cases failed and when localized beyond the curvature 15 of 33 cases failed. Anatomical factors favorable for removal were: straight canals, incisors and canines, localization before the curvature, length of fragment more than 5 mm, localization in the coronal or mesial third of the root canal, reamer or lentulo spirales. In molars removal procedures were most successful in the palatal canals of maxillary molars. PMID- 10825836 TI - Clinical evaluation of glutaraldehyde with calcium hydroxide and glutaraldehyde with zinc oxide eugenol in pulpotomy of primary molars. AB - The objectives of this study were to clinically evaluate the effectiveness of 2% buffered glutaraldehyde in pulpotomies of human primary molars and to compare the success rate of glutaraldehyde with calcium hydroxide and glutaraldehyde with zinc oxide eugenol as dressing material on the radicular pulp. Pulpotomies were completed on 61 primary molars in 19 children. The teeth were divided into two groups by random allocation. One group had a dressing of zinc oxide-eugenol base (IRM) incorporated with one drop of 2% buffered glutaraldehyde while the other group had a dressing of calcium hydroxide base incorporated with one drop of 2% buffered glutaraldehyde after the initial placement of 2% buffered glutaraldehyde on cotton pellet for 3 min. All teeth had light-curing compomer (Dyract) placed over the dressing material followed by a stainless steel crown restoration within 1 or 2 weeks after the pulpotomy. Blind clinical and radiographic evaluations of 57 teeth available after 12 months showed a success rate of 92.9% and 73.6% respectively. The clinical and radiographic success rates for glutaraldehyde/zinc oxide eugenol pulpotomies were 96.5% and 75.8% respectively while those for glutaraldehyde-calcium hydroxide were 89.2% and 71.4%. There was no statistically significant difference between the two groups either clinically or radiographically. The overall clinical success rate suggested that 2% buffered glutaraldehyde was an effective agent in the pulpotomy of human primary molars. PMID- 10825837 TI - Ultrasonic root-end preparation with smooth and diamond-coated tips. AB - The purpose of this study was to compare the effects of smooth and diamond-coated ultrasonic retrotips on the external and internal surfaces of root-end preparations with the aid of a scanning electron microscope (SEM). Forty-four mesial roots of human mandibular molars were selected. The canals were cleaned, shaped and obturated using gutta-percha and sealer. The apical portions were resected at a 45 degrees-angle bevel exposing both mesial canals and the isthmus area. The roots were then divided into two groups according to the type of root end preparation: Group A--performed with smooth retrotips (S) and Group B- performed with diamond-coated retrotips (DC). The specimens were coded and prepared for SEM evaluation. Observations of the external surface preparation showed that the S and DC retrotips produced very well-centered cavities involving both canals and isthmus area with minimal deviations and no perforative defects. When the internal surface of the root-end preparations was evaluated, it was evident that the use of S retrotips resulted in clean canal walls with little superficial debris and smear layer. Internal canal surfaces done with DC retrotips were irregular showing patent grooves, in contrast with the more uniform, regular and smoother surfaces when S retrotips were employed. PMID- 10825838 TI - Inflammatory and replacement resorption in reimplanted permanent incisor teeth: a study of the characteristics of 84 teeth. AB - Avulsion is a serious injury which can cause damage to some or all of the dental and surrounding tissues. This study examined the profiles of teeth showing inflammatory resorption, in terms of time prior to reimplantation, contamination, pulp extirpation time and period of splinting and compared them to teeth without resorption. There were a total of 71 children in the present study (mean age 9.8 years, range 6-16 years) with a total of 84 reimplanted teeth. Inflammatory resorption was present in 22 teeth. There was a significant relationship between the presence of inflammatory resorption and the time the teeth were dry prior to reimplantation, with a lesser effect for total delay time. There were slightly later pulp extirpation times for teeth with inflammatory resorption, with median delays of 16 and 11 days respectively and increased inflammatory resorption in teeth extirpated at 20 days or later. Replacement resorption was present in 40 teeth. There was a significantly longer splinting time in teeth with replacement resorption and more resorption in teeth splinted for longer than 10 days. It was concluded that pulp extirpation time was not critical unless the delay exceeded 20 days and that splinting time should not exceed 10 days. PMID- 10825839 TI - Non-vital immature permanent incisors: factors that may influence treatment outcome. AB - This study examines the treatment of non-vital immature permanent incisors using the calcium hydroxide apexification technique. The objectives of the present study were to determine the speed and location of barrier formation and those factors discernible at presentation and during treatment which may be related to it. Forty-four non-vital immature incisors undergoing calcium hydroxide apexification were reviewed in detail. All cases were reviewed every 8-12 weeks for up to 18 months, or until apexification occurred. Details of the time and nature of the injuries and treatment were recorded. The degree of apical development prior to treatment was assessed, and barrier formation, location and time were noted. Mean time to barrier formation was 34.2 weeks (range 13-67 weeks). The strongest predictor of rapid barrier formation was the rate of change of calcium hydroxide and a barrier also formed more rapidly in cases with narrower initial apical width. There was evidence of displacement and a higher mean time for barrier formation in half of the cases. The presence of an abscess was the weakest predictor of rapid barrier formation and the effect was not significant (P = 0.280). The barrier was located at the apex in 28 cases (63.6%) and the distance from the barrier to the apex for the remaining 16 (36.4%) varied from 1 mm to 5 mm. The number of placements of calcium hydroxide varied from 1 to 4 with a mean of 1.9, and there was a higher mean number of calcium hydroxide placements in the cases where the barrier was located at the apex. PMID- 10825840 TI - The diagnostic accuracy of microbiologic root canal sampling and the influence of antimicrobial dressings. AB - The routine approach to endodontic treatment of teeth with apical periodontitis often involves an interappointment dressing with calcium hydroxide. However, investigations have demonstrated a negative influence of calcium hydroxide on the accuracy of microbiological root canal sampling (MRS). The aim of the present study was to investigate whether the use of a fluid dressing like 5% iodine potassium iodide (IPI) would increase the accuracy of MRS. Following instrumentation of 50 teeth with radiographically verified apical periodontitis the root canals received IPI as an intracanal dressing. One week after closure canals were sampled, "test sample" (TS), and then left filled with sampling fluid and temporarily scaled. Seven days later a "gold standard" (GS) sample was obtained. Bacteria were recovered in 22 teeth (44%) in TS as well as in GS. Fifteen teeth (30%) were positive for growth in both samples. Using the detection level "very sparse growth" of microbes the sensitivity and specificity of MRS reached 68% and 75%, respectively. In an earlier study, following the same experimental protocol, but with calcium hydroxide as intracanal dressing, the corresponding values were 33% and 81%. In 25% of these cases bacteria persisted in the canals. As compared to calcium hydroxide, the use of IPI resulted in improved test accuracy, but loss of antibacterial capacity. Conclusively, intracanal dressings seem to vary in their influence on the microbiologic test performance as well as in their antibacterial efficacy. In a clinical situation the choice of interappointment dressing should include consideration of these potentially conflicting properties. PMID- 10825841 TI - A comparison of laser Doppler flowmetry with other methods of assessing the vitality of traumatised anterior teeth. AB - Laser Doppler flowmetry is a non-invasive electrooptical technique which allows the semi-quantitative recording of pulpal blood flow. This study aimed to determine the reliability (measured as the sensitivity and specificity) of laser Doppler flowmetry as a method of assessing the vitality of traumatised anterior teeth, and to compare it with standard pulpal diagnostic tests. Recordings of pulpal blood flow were taken from 67 non-vital anterior teeth (55 patients), where the pulpal status was confirmed by pulpectomy. For comparison, recordings were also taken from 84 vital anterior teeth (84 patients). Analysis of the recordings allowed diagnostic criteria to be developed which gave the technique a sensitivity and specificity of 1.0 for this sample. None of the other standard pulpal diagnostic methods tested was as reliable. This was usually due to low sensitivities, which ranged between 0.92 for sensibility testing with ethyl chloride down to 0.36 for periapical radiolucency and 0.16 for a history of pain. Laser Doppler flowmetry was found to be a reliable method of assessing the pulpal status of traumatised anterior teeth, although it is technique-sensitive and time consuming to use. PMID- 10825842 TI - Mental nerve paresthesia associated with an amalgam filling: a case report. AB - We present a case report in which a mental nerve paresthesia occurred in connection with an amalgam filling placed in direct contact with the pulp of a mandibular first molar. The main evidence for amalgam being the dominant etiological factor was the positive epicutaneous patch testing response to the components of the amalgam alloy, and the total remission of symptoms without further clinical complications after removal of the amalgam filling and subsequent endodontic therapy. PMID- 10825843 TI - Malformation in the primary and permanent dentitions following trauma prior to tooth eruption: a case report. AB - Dento-facial injuries that occur prior to the eruption of teeth in the primary dentition are rare, but can result in damage to the primary dentition. We report a rare case where an injury to the anterior maxilla and mandible of an infant prior to primary tooth eruption resulted in hypoplasia, displacement and impaction of the primary dentition and damage to a developing permanent tooth. PMID- 10825844 TI - Mobility in old age. AB - Immobility is common in older people and may impact on their dental care. Immobility in old age may have physical, psychological and environmental causes. Immobile elderly people often suffer from a number of diseases which worsen their mobility. Arthritis, osteoporosis, hip fracture, stroke and Parkinson's disease are among the most common causes of immobility in old age. Complications of immobility such as orthostatic hypotension may occur in the dental patient. Careful history-taking and a thorough physical examination by the physician are the most important parts of the assessment process. This assessment should lead to a list of active problems and treatment should then be aimed at these problems. Active management, carried out by the multidisciplinary team, will lead to improvements in mobility and lessen the frequency and severity of the complications of immobility. This broad description thus provides the basis for a wide understanding for the special problems that the immobile patient present to the practitioner and ways of overcoming the problems. PMID- 10825845 TI - Behind the screens: care staff observations on delivery of oral health care in nursing homes. AB - OBJECTIVE: To identify qualitatively carer staff attitudes, practices and critical comments related to oral health care of functionally dependent nursing home clients. DESIGN: Open-ended questions included in a longer quantitative questionnaire. SETTING: 22 randomly selected nursing homes in the Bristol area. SUBJECTS: 416 carers employed in these homes. RESULTS: The majority of carers thought that clients had a right to good oral health, accepted the carers' role in helping clients with oral and denture hygiene, but recognised that oral health care provision was deficient. However, some carers believed oral health care to be solely the clients' responsibility despite high disability levels. Main barriers to providing oral health care were low prioritization of oral health by nursing management, lack of co-operation from cognitively impaired clients, and lack of training. Carers were critical of homes' lack of arrangements for routine professional dental checks, lack of commitment to staff training, low standards of oral health care by colleagues, and lack of provision of oral hygiene aids and cleansing materials for clients. Many responses indicated ways in which nursing home oral health care could be improved. Carers reported contrasting experiences of dental treatment, and deplored recent decreased availability of subsidized dental care. CONCLUSIONS: Carers' generally positive attitudes towards clients' oral health care should encourage health educators. Insights gained from qualitative data can help to identify the less obvious causes for poor oral care delivery, which can then be addressed in education and training initiatives in nursing home settings. PMID- 10825846 TI - The orodental status of a group of elderly in-patients: a preliminary assessment. AB - OBJECTIVE: To provide a preliminary assessment of the orodental status and dental treatment requirements of a group of elderly in-patients. DESIGN: Cross sectional. SETTING: Acute Care of the Elderly and Stroke Rehabilitation units at teaching hospitals in Merseyside. SUBJECT: 150 patients aged 58 to 94 years, in which a history could be validated at interview. INTERVENTION: Questionnaire administered by dentist and clinical examination. MAIN OUTCOME MEASURES: Registration with a dentist, prosthetic status and difficulties with dentures, denture hygiene and identification marking, dental treatment needs and evidence of mucosal pathology. RESULTS: Only 27% of patients claimed registration with a dentist. Three quarters of the patients were edentulous and 66 patients wore full dentures; 18 had no prostheses. Difficulties were experienced by one quarter of patients with upper dentures, compared with a half of lower denture wearers. Of the dentures available for inspection, 61% had removable soft debris, 66% were left out at night and 75% were cleaned by the patient, whilst on the ward. No dentures had evidence of identification marking. Of the 39 partially dentate patients, 75% required interventive dental treatment. Denture stomatitis was diagnosed in 29% of patients and 19 had evidence of benign mucosal pathology. CONCLUSIONS: The orodental status of this group of elderly in-patients was poor, with a high proportion being edentulous. Few were registered with a dentist and denture hygiene was inadequate. Lack of identification marking is a matter of concern. Closer liaison between hospital staff responsible for elderly in patients is required, to improve the orodental health and quality of life of this medically compromised group of patients. PMID- 10825847 TI - An index of clinical oral disorder in elders (CODE). AB - Attempts to construct an index of oral health or disorder suitable for older adults have been limited in clinical scope or based on the judgement of very few individuals. Consequently, we present here a multidimensional index of Clinical Oral Disorder in Elders (CODE) based on a breadth of clinical measures relevant to elders. The data for the index are derived from a clinical examination of jaw function, dentures, mucosa, teeth, and periodontium. Weighting or ranking for each disorder within the context of an elderly person was established as mild, moderate or severe by experienced dental clinicians and dental hygienists. Subsequently, the index was constructed by transforming the weights into a numerical value for each clinical measure. Individual scores can relate to the heaviest weight identified during the examination or to the sum of the weights assigned throughout the examination, and the average score during a series of examinations will reflect the clinical status of a particular population. Clinical applications in elderly residential-care populations indicate that the index can be constructed efficiently and with reasonable reliability. We suggest, therefore, that the CODE index is suitable for descriptive and comparative research by providing a clinical format for measuring oral disorder in disabled elders, and, when combined with a psychosocial index, it should be very comprehensive indicator of oral dysfunction in older adults. PMID- 10825848 TI - Factors influencing older people's self reported use of dental services in the UK. AB - OBJECTIVES: This study was designed to determine the use of dental services and factors associated with their use among the United Kingdoms' older population. DESIGN: A national study involving 1,116 older people (aged 60 or older). SETTING: Home Interviews were undertaken exploring the time and reason for last dental visit. In addition, socio-demographic characteristics and proxy oral health measures (self-reported number of teeth and edentulous status) of the respondents were collected. RESULTS: Forty seven percent (528) claimed they visited the dentist within the past year, 10% (116) claimed that the reason for their last visit was because of a dental emergency, 43% (484) were classified as "regular attenders"--having attended the dentist within the past year for a non dental emergency. Bivariate analysis identified that regular dental attendance was associated with age (P < 0.01), social class (P < 0.01), income level (P < 0.01), educational attainment (P < 0.01), self-reported number of teeth possessed (P < 0.01) and edentulous status (P < 0.01). In regression analysis, self reported edentulous status and number of teeth possessed emerged as the most important factors in determining service utilisation. Possessing a full denture was associated with a 6-fold decrease, having accounted for other factors, in the likelihood of attending the dentist within the past year for a non dental emergency (OR = 0.15, CI 0.10, 0.21). CONCLUSION: Less than half of the sample population were "regular dental attenders", their attendance was associated with a number of socio-demographic and oral health factors. In particular, edentulous state was a major factor associated with their use of services. PMID- 10825849 TI - The oral health of older people from minority ethnic communities in south east England. AB - OBJECTIVES: To describe the clinical health status and subjective oral health status of older adults from minority ethnic communities resident in South East England. DESIGN: Cross sectional survey incorporating a clinical examination and a questionnaire assessment of subjective oral health status. SETTING: Community groups working with individuals from minority ethnic communities. SUBJECTS: A total of 540 individuals from 7 minority ethnic communities. MEASURES: Clinical assessment of oral health status using BASCD criteria. Assessment of oral symptoms and impact experienced together with satisfaction with oral status, by structured questionnaire. RESULTS: On all measures of clinical health status the participants were healthier than a comparison group based on data from the Adult Dental Health Survey. The participants expressed high levels of subjective oral symptoms. Levels of satisfaction were lower than those found in the Adult Dental Health Survey. There was evidence of some variation across minority ethnic communities in clinical and subjective oral health status. CONCLUSIONS: Approximately 20% of the 540 adults from minority ethnic communities resident in the South East of England surveyed were relatively fit orally. They experienced a great many oral symptoms, and in approximately 30% of individuals these were sufficient to interfere with their quality of life. ETHICAL APPROVAL: The research described in this paper was approved by the Ethics committee of King's College London. PMID- 10825850 TI - Efficacy of antifungal agents in tissue conditioners in treating candidiasis. AB - Chronic atrophic candidiasis is prevalent in up to 72% of institutionalized geriatric populations and is causally associated with Candida albicans. Topical antifungal treatments are difficult to implement in some geriatric patients due to cognitive impairment, reduced motor dexterity and memory loss. OBJECTIVE: This in vitro study incorporated antifungal agents into tissue conditioners to investigate the effectiveness of this method of drug delivery. DESIGN: Combinations of nystatin, fluconazole, itraconazole and Coe Soft, Viscogel, Fitt were tested at 1, 3, 5, 7, 9 and 11 wt/wt%, with and without sterilized saliva. 6 mm diameter cores were punched in Sabouraud plates pre-grown with standardized C. albicans. Antifungal agents plus tissue conditioner mixtures were injected into each core. Inhibition diameters were measured for 14 days. RESULTS: Cores with only tissue conditioners acted as negative control and showed no significant inhibition activity (ANOVA, p > 0.05). Peak activity was between 65 to 89 hours; followed by a plateau. Itraconazole had greater fungicidal activity than fluconazole; while nystatin was found to have the least fungicidal activity (ANOVA, p < 0.05). The most effective concentration for nearly all combinations was 5% wt/wt (ANOVA, p < 0.05). Specimens with saliva showed greater antifungal activity than those without (t-test, p < 0.001). Itraconazole altered the physical properties of Viscogel hence this combination is not recommended for clinical use. CONCLUSION: The treatment of chronic atrophic candidiasis by incorporation of antifungal drugs into tissue conditioners is efficacious. 5% wt/wt itraconazole mixed with Coe Soft or Fitt is recommended for clinical study where compliance of patient or care giver cannot be relied upon. Peak antifungal activity at 3 days suggests that mixtures prepared for clinical study may be replaced soon after this time for maximum effectiveness. PMID- 10825851 TI - Bitter taste thresholds, numbers and diameters of circumvallate papillae and their relation with age in a Turkish population. AB - OBJECTIVES: This study analyses the relationship between the sense of bitter taste and age. The relationships between these and the numbers and diameters of Circumvallate Papillae (CP) are also analysed. SUBJECTS: Twenty-four elderly subjects (from 65 to 85 years) and 30 young subjects (from 17 to 25 years) were studied. METHOD: Bitter taste thresholds were determined by the three drop method with an ascending series of concentrations. The numbers and diameters of CP were observed by direct naked eye observation using a wooden tongue depressor, a gauge and a light source. RESULTS: The bitter taste acuity was significantly poorer in the older sample. There was no relationship between the bitter taste acuity and number of papillae but an inverse relationship was observed between the bitter taste acuity and diameters of papillae, thus lower acuity was associated with larger papillae. It was also observed that the diameters of papillae were inversely related to the numbers of CP. The distribution of numbers and diameters of CP were not significantly different between these samples with age. PMID- 10825852 TI - An abstract: Technique and philosophy for the treatment of Class II skeletal and dental malocclusions. PMID- 10825853 TI - A new perspective on the methodology for clinical efficacy evaluations of tooth whitening dentifrices. PMID- 10825854 TI - A six-week clinical efficacy study of a tooth whitening tartar control dentifrice for the removal of extrinsic tooth stain. AB - The objective of this double-blind clinical study was to investigate the tooth whitening efficacy of a new commercially available calculus-inhibiting dentifrice (Colgate Tartar Control Plus Whitening Fluoride dentifrice) containing tetra sodium pyrophosphate, sodium tripolyphosphate, and PVM/MA copolymer in a 0.243% sodium fluoride/silica base (test dentifrice), compared to a commercially available, non-tooth whitening dentifrice containing 0.243% sodium fluoride in a silica base (control dentifrice). Following a baseline examination for, extrinsic tooth stain, qualifying adult male and female subjects from the Manchester, United Kingdom area were randomized into two treatment groups which were balanced for gender, age and level of extrinsic tooth stain. Subjects were instructed to brush their teeth twice daily (morning and evening) for one minute with their assigned dentifrice using a soft-bristled toothbrush. Examinations for extrinsic tooth stain were repeated after six weeks' use of the study dentifrices. Seventy nine(79) subjects complied with the protocol and completed the entire study. At the six-week examination, subjects assigned to the test dentifrice treatment group exhibited statistically significant reductions of over 40% with respect to both extrinsic tooth stain area and extrinsic tooth stain intensity relative to those subjects assigned to the control dentifrice treatment group. PMID- 10825855 TI - A six-week clinical efficacy study of a new dentifrice for the removal of extrinsic tooth stain. AB - The objective of this double-blind clinical study was to investigate the tooth whitening efficacy of a new dentifrice (Colgate Tartar Control Plus Whitening Fluoride Toothpaste) for the removal of extrinsic tooth stain, through a comparison with two commercially available dentifrices, Aquafresh Whitening Toothpaste with Fluoride, and Crest Regular Fluoride Toothpaste. Following a baseline examination for extrinsic tooth stain area and intensity, qualifying adult male and female subjects from the Mississauga, Ontario, Canada area were randomized into three treatment groups which were balanced for age, gender, and level of extrinsic tooth stain. Subjects were instructed to brush their teeth twice daily (morning and evening) for one minute with their assigned dentifrice using a soft-bristled toothbrush. Examinations for extrinsic tooth stain were repeated after six weeks' use of the study dentifrices. One hundred twenty-two (122) subjects complied with the protocol and completed the entire study. At the six-week examination, subjects assigned to the Colgate Tartar Control Plus Whitening treatment group and subjects assigned to the Aquafresh Whitening treatment group exhibited statistically lower levels (p < 0.0001) of extrinsic tooth stain area and intensity than did those subjects assigned to the Crest Regular Fluoride treatment group. Further, subjects assigned to the Colgate Tartar Control Plus Whitening treatment group exhibited significantly lower levels of extrinsic tooth stain area and intensity than did those assigned to the Aquafresh Whitening treatment group (p < 0.0001). PMID- 10825856 TI - The clinical anticalculus efficacy of a tartar control whitening dentifrice for the prevention of supragingival calculus in a three-month study. AB - The objective of this double-blind clinical study was to compare the effect of a new dentifrice (Colgate Tartar Control Plus Whitening Fluoride Toothpaste) for the prevention of supragingival calculus, with that of a commercially available calculus-inhibiting dentifrice (Crest Tartar Control Toothpaste). The study involved adult male and female subjects who had pre-qualified for participation by developing sufficient supragingival calculus (greater than 7.0 on the Volpe Manhold Calculus Index) during an eight-week screening period. Subjects received a full oral prophylaxis, and were stratified into two treatment groups balanced for age, sex and qualifying calculus score. Subjects were instructed to brush their teeth twice daily (morning and evening) for one minute with their assigned dentifrice using a soft-bristled toothbrush. Examinations for dental calculus were performed after twelve weeks' use of the study dentifrices, using the Volpe Manhold Calculus Index, Fifty-eight (58) subjects complied with the protocol and completed the entire study. The Colgate Tartar Control Plus Whitening group exhibited a statistically significant (p < 0.001) 34.6% reduction in mean calculus score compared to the Crest Tartar Control group. PMID- 10825857 TI - A six-week clinical efficacy study of three commercially available dentifrices for the removal of extrinsic tooth stain. AB - The objective of this double-blind clinical study was to compare the extrinsic tooth stain removal efficacy of three commercially available dentifrices: Colgate Platinum Whitening Toothpaste with Fluoride, Aquafresh Advanced Whitening Toothpaste with Fluoride, and Crest Regular Fluoride Toothpaste. Following a baseline examination for extrinsic tooth stain, qualifying adult male and female subjects from the Toronto, Ontario, Canada are were randomized into three treatment groups which were balanced for age, gender, tobacco habits, and level of extrinsic tooth stain using the Lobene Stain Index. Subjects were instructed to brush their teeth twice daily (morning and evening) for one minute with their assigned dentifrice using a soft-bristled toothbrush. Examinations for extrinsic tooth stain were repeated after six weeks' use of the study dentifrices. One hundred forty-nine (149) subjects complied with the protocol and completed the entire study. At the six-week examination, subjects assigned to the Colgate Platinum treatment group exhibited statistically significant reductions (p < 0.05) from baseline level with respect to extrinsic tooth stain area (49.4% reduction) and extrinsic tooth stain intensity (52.4% reduction). Statistically significant reductions (p < 0.05) for both extrinsic tooth stain parameters were also exhibited by the subjects assigned to the Aquafresh Whitening treatment group (14.0% reduction for area, 15.5% reduction for intensity). Both tooth whitening treatments provided statistically significantly greater reductions (p < 0.05) in extrinsic tooth stain than did the Crest Regular Fluoride dentifrice, which exhibited a small increase in the extrinsic tooth stain parameters over the course of the study. Additionally, the Colgate Platinum dentifrice provided statistically significantly (p < 0.05) greater removal of extrinsic tooth stain over the course of this six-week study than did the Aquafresh Whitening dentifrice. Thus, the results of the double-blind clinical study support the conclusion that the two tooth whitening dentifrices, Colgate Platinum and Aquafresh Whitening, provide significantly greater (p < 0.05) removal of extrinsic tooth stain than does Crest Regular Fluoride, a sodium fluoride/silica dentifrice. Further, the results of the study support the conclusion that Colgate Platinum dentifrice provides a greater (p < 0.05) level of extrinsic tooth stain removal efficacy than does Aquafresh Whitening dentifrice. PMID- 10825858 TI - A six-week clinical efficacy study of four commercially available dentifrices for the removal of extrinsic tooth stain. AB - The objective of this double-blind clinical study was to compare the efficacy for extrinsic tooth stain removal of four commercially available dentifrices: Colgate Tartar Control with Baking Soda & Peroxide Fluoride Toothpaste; Aquafresh Advanced Whitening Toothpaste with Fluoride; Rembrandt Tartar Control Low Abrasion Fluoride Whitening Toothpaste; and Crest Regular Fluoride Toothpaste. Following a baseline examination for extrinsic tooth stain on the anterior six mandibular and maxillary teeth, qualifying adult male and female subjects from the Philadelphia, Pennsylvania area were randomized into four treatment groups which were balanced for age, gender, tobacco habits, and level of extrinsic tooth stain. Subjects were instructed to brush their teeth twice daily (morning and evening) for one minute with their assigned dentifrice using a soft-bristled toothbrush. Examinations for extrinsic tooth stain were repeated after six weeks' use of the study dentifrices. One hundred eighty (180) subjects complied with the protocol, and completed the entire study. At the six-week examination, subjects assigned to the Colgate Tartar Control with Baking Soda & Peroxide Fluoride Toothpaste treatment group exhibited statistically lower levels of extrinsic tooth stain area and extrinsic tooth stain intensity than did those subjects assigned to the Crest Regular Fluoride treatment group. Subjects assigned to the Aquafresh Advanced Whitening treatment group exhibited significantly lower levels of extrinsic tooth stain area than did those assigned the Crest Regular Fluoride group. No other significant differences among the four study dentifrices were noted. Thus, the results of this double-blind clinical study support the conclusion that Colgate Tartar Control with Baking Soda & Peroxide Fluoride Toothpaste provides significantly greater control of extrinsic tooth stain than does Crest Regular Fluoride, a sodium fluoride/silica dentifrice. PMID- 10825859 TI - A short-term brushing model for assessing antiplaque/antigingivitis dentifrice effectiveness: a pilot study. AB - This pilot clinical study was conducted to evaluate the utility of a short-term brushing model for assessing the inhibition of supragingival plaque and gingivitis by an essential oil-containing dentifrice formulation. Forty-two qualifying subjects entered and completed this double-blind, parallel group, controlled study. At baseline, subjects received an oral soft and hard tissue examination and were also scored for plaque, gingivitis and bleeding, both visually and invasively. Subjects received a supragingival prophylaxis and were randomly assigned to an experimental essential oil-containing dentifrice or its vehicle control. Subjects brushed unsupervised for one minute, twice daily for three weeks, after which all clinical examinations were repeated. All examinations were performed by a qualified and calibrated examiner. Analyses of covariance were used to compare inter-group means for each of the three outcome variables. Compared to its vehicle control, the experimental dentifrice produced statistically significant (p < 0.0001) reductions of 39.6% for plaque, 10.8% for gingivitis, and 65.4% for bleeding. No adverse events were observed or reported. The results of this pilot study suggest that three weeks of conventional use may be an adequate duration to demonstrate effectiveness of an essential oil containing dentifrice. Confirmation of these results in a duplicate study is desirable to assess the reliability of the model. Furthermore, the utility of this model to determine the effects of formula modifications to the antiplaque and antigingivitis activity of active agents needs to be explored. PMID- 10825860 TI - Microleakage of Class V restorations using two different compomer systems: an in vitro study. AB - This study compared the marginal microleakage of Class V cavities restored with Dyract-AP and F2000. Forty Class V cavity preparations were performed on extracted human teeth. As a negative control, twenty teeth were used without Class V preparations. The apical foramina of the teeth were sealed with a layer of varnish and amalgam restorations. Class V cavity preparations with occlusal margins in enamel, and gingival margins in cementum or dentin that measured approximately three millimeters in width (gingival-occlusal) and length (mesial distal), were prepared on the buccal surface of the teeth. Samples were divided randomly into 2 groups of 15, and restored per manufacturer's instructions using experimental primer/conditioner (PCC, D/C) and Single Bond Adhesive (3M). All restorations were polished with an abrasive finishing kit. After storage in 37 degrees C water for 24 hours, all specimens were thermocycled between 5 degrees C and 55 degrees C for 500 cycles with a 30-second dwell time, followed by immersion in 0.2% basic fucsine for 24 hours. Teeth were then embedded in cold cure acrylic resin, sectioned longitudinally, and the dye penetration at the enamel and cementum margins were scored at 30x magnification. Evaluations were rated from 0 to 3 (0 = no leakage; 1 = dye penetration up to one-half of the preparation depth; 2 = dye penetration more than one-half preparation depth, but less than the axial wall; 3 = dye penetration along the axial wall). Both F-2000 and Dyract-AP indicated no leakage in the enamel margins. Dyract-AP showed no leakage at either the enamel or dentin margins. Fisher's Exact Test revealed that this difference in dentin margins was statistically significant (p < 0.05). Under the given conditions, Dyract-AP and F-2000 demonstrated resistance to microleakage in enamel, and showed Dyract-AP to be more resistant to microleakage in dentin than F-2000. PMID- 10825861 TI - Plaque removal efficacy of the V-shaped toothbrush with new designs in bristle arrangement. AB - Experimental bristle arrangements were applied to a conventional V-shaped bristle design toothbrush in an effort to improve plaque removal efficacy in interproximal areas. A single-blind cross-over study was performed to evaluate the plaque removal efficacy of this new bristle arrangement by comparing it to either a more conventional V-shaped toothbrush or a flat-headed toothbrush. Plaque removal efficacy was determined by assessing the percentage of plaque score reduction following a single controlled brushing. The new toothbrush bristle arrangement had a significantly higher plaque removal percentage efficacy than both the V-shaped toothbrush (59.1% vs. 48.5%; p = 0.0092) and the flat headed toothbrush (65.3% vs. 55.3%; p = 0.0260) in interproximal areas. These differences were also consistent with whole mouth comparisons. When the subjects were asked about their preferences for the three different bristle formats used in this study, there was no significant difference of opinion found. These findings indicated a superiority of the new toothbrush to the other two, more conventional toothbrush bristle styles on plaque removal efficacy with these study subjects, but no particular preference for bristle design. PMID- 10825862 TI - The clinical effectiveness of a dentifrice containing triclosan and a copolymer for controlling breath odor measured organoleptically twelve hours after toothbrushing. AB - The objective of this double-blind clinical study was to investigate the effectiveness of a dentifrice containing triclosan and a copolymer, Colgate Total Toothpaste, for controlling breath odor twelve hours after brushing the teeth compared to a placebo dentifrice which did not contain triclosan or a copolymer. Breath odor was evaluated by a panel of four expert judges using a nine-point hedonic scale. Following a baseline evaluation of breath odor, prospective study subjects who presented scores above the threshold value for unpleasant breath odor were stratified by score, and randomized into two treatment groups. Subjects were provided with a soft-bristled toothbrush, and brushed their teeth thoroughly in their regular and customary manner with their assigned dentifrice. Subjects refrained from dental hygiene, breath mints, or mouth rinses for the next twelve hours, after which they were once again evaluated for breath odor. Sixty-three (63) adult male and female subjects from the Mississauga, Ontario, Canada area participated in the study. At twelve hours after brushing their teeth, subjects in both dentifrice treatment groups presented mean breath odor scores which were statistically significantly lower than the mean scores observed at baseline. However, the difference between the mean twelve-hour breath odor scores differed significantly between the two dentifrices. The mean twelve-hour breath odor score for the Colgate Total Toothpaste group was 4.77, which is within the range of values corresponding to pleasant breath odor; the mean twelve-hour breath odor score for the placebo group was 6.05, which is above the value corresponding to unpleasant breath odor. Thus, the results of this double-blind clinical study support the conclusion that Colgate Total Toothpaste provides effective control of breath odor at twelve hours after brushing the teeth. PMID- 10825864 TI - Subgingival access and artificial plaque removal by a sonic cleaning device. AB - A sonic subgingival cleaning device (soniPick Sonic Interdental Plaque Remover) has recently been marketed with three bristle tips varying in size lengths and bundle diameters. The purpose of this study was to evaluate the ability of these bristle tips to remove subgingival artificial plaque deposits in a laboratory method used for toothbrushes. The laboratory method has been modified for testing the sonic device, to simulate the directions for using the product at home. The dimensions (length x bundle width) of three tips tested were: 0.007" x 0.040"; 0.007" x 0.062"; and 0.009" x 0.062". The tips used with the sonic device were placed on the tooth surface at a 20 degrees angle according to directions, and inserted 3 mm under simulated gingivae. The device was turned on and the tip was moved in a maximum 10 mm stroke for 15 seconds with the brushing machine. For control purposes, a flat, multi-tufted, ADA-accepted manual toothbrush (Oral-B P35) was also tested. The manual toothbrush was tested using the standard methods for evaluating toothbrushes, i.e., with the bristle tips placed at the gingival margin, then brushed at a 45 degrees angle at 250 g weight, using 15 mm strokes for 60 seconds. The depth of subgingival deposit removal was recorded as the maximum depth of the artificial plaque deposit removed from the pressure sensitive paper under the simulated gingivae over anterior- or posterior-shaped teeth. In the assays conducted, the three bristle tips on the sonic device removed artificial plaque deposits under the simulated gingival at depths of 1.9 2.7 mm. The manual toothbrush had a mean subgingival cleaning depth of 0.6 mm. Differences between the three bristle tips used with the sonic device and the manual toothbrush were significant (p < 0.001 ANOVA). In this laboratory assay, all 3 bristle tips provided with the sonic cleaning device maintained access into and subsequent removal of artificial plaque from the subgingival space. PMID- 10825863 TI - The clinical effectiveness of a dentifrice containing triclosan and a copolymer for providing long-term control of breath odor measured chromatographically. AB - The objective of this double-blind clinical study was to investigate the effectiveness of a commercially available dentifrice containing triclosan and a copolymer (Colgate Total Toothpaste) for controlling long-term, i.e., seven-hour and overnight breath odor. In particular, a comparison was made between the level of control of breath odor provided by the test dentifrice, and that provided by a placebo dentifrice which did not contain triclosan or a copolymer. This study followed a two-treatment, two-period crossover design. Prospective subjects were provided with a supply of a commercially available fluoride dentifrice, which was used for a one-week period prior to the two seven-day treatment periods. During each treatment period, subjects were instructed to brush their teeth twice a day, morning and evening, for sixty seconds with their assigned study dentifrice, using the soft-bristled toothbrush which had been provided. On the morning following the seventh day of each treatment period, subjects reported to the clinical facility for overnight breath odor assessments. Directly following this, subjects brushed their teeth, ate and drank normally, and reported once again to the clinical facility at seven hours post-toothbrushing for another breath odor assessment. Prior to the overnight breath odor assessments, subjects refrained from brushing their teeth, rinsing their mouths or using breath mints, and from eating or drinking anything on the morning of the evaluation. Subjects refrained from the use of tobacco products, and from eating onions, garlic, or strong spices throughout the entire study. Breath odor was instrumentally evaluated by measuring the level of volatile sulfur compounds in the mouth air using a 565 Tracor gas chromatograph equipped with a flame photometric detector. Measurements were taken in duplicate, and then averaged. Levels of volatile sulfur compounds were expressed in nanograms per milliliter (ng/ml) of mouth air. The two dentifrices exhibited statistically significant differences (p < 0.05) with respect to both overnight breath odor and seven-hour post-toothbrushing breath odor. The mean overnight breath odor scores were 9.63 ng/ml for Colgate Total Toothpaste, and 12.64 ng/ml for the placebo dentifrice. For seven-hour breath odor, the mean scores were 5.62 ng/ml for Colgate Total Toothpaste, and 7.10 ng/ml for the placebo dentifrice. Thus, the results of this double-blind clinical study on 19 subjects support the conclusion that Colgate Total Toothpaste provides effective seven-hour and overnight control of breath odor. PMID- 10825865 TI - Dental discoloration: an overview. AB - Often the first evidence of variation from normal in human dentition is an observable difference in the color of the teeth. During the past decade, the demand for conservative esthetic dentistry has grown dramatically. Tooth discoloration is a frequent dental finding, associated with clinical and esthetic problems. It differs in etiology, appearance, composition, location, severity, and firmness in adherence to the tooth surface. Basically, there are two types of tooth discolorations: those caused by extrinsic factors and those caused by intrinsic congenital or systemic influence. The intensity of stains may be worsened if there are enamel defects. Tooth discoloration presents two major challenges to the dental team. The first challenge is to ascertain the cause of the stain; the second is its management. CLINICAL SIGNIFICANCE: This article reviews the etiology and clinical presentation of dental stains and outlines treatment options. PMID- 10825866 TI - Comparing the perception of dentists and lay people to altered dental esthetics. AB - PURPOSE: This study was designed to determine the perceptions of lay people and dental professionals with respect to minor variations in anterior tooth size and alignment and their relation to the surrounding soft tissues. MATERIALS AND METHODS: Smiling photographs were intentionally altered with one of eight common anterior esthetic discrepancies in varying degrees of deviation, including variations in crown length, crown width, incisor crown angulation, midline, open gingival embrasure, gingival margin, incisal plane, and gingiva-to-lip distance. Forty images were randomized in a questionnaire and rated according to attractiveness by three groups: orthodontists, general dentists, and lay people; 300 questionnaires were distributed. RESULTS: The response rate was 88.2% for orthodontists, 51.8% for general dentists, and 60.6% for lay people. The results demonstrated threshold levels of noticeable difference between the varying levels of discrepancy. A maxillary midline deviation of 4 mm was necessary before orthodontists rated it significantly less esthetic than the others. However, general dentists and lay people were unable to detect even a 4-mm midline deviation. All three groups were able to distinguish a 2-mm discrepancy in incisor crown angulation. An incisal plane cant of 1 mm as well as a 3-mm narrowing in maxillary lateral incisor crown width were required by orthodontists and general dentists to be rated significantly less esthetic. Lay people were unable to detect an incisal plane asymmetry until it was 3 mm, or a lateral incisor narrowing until it reached 4 mm. Threshold levels for open gingival embrasure and gingiva-to-lip distance were both at 2 mm for the orthodontic group. Open gingival embrasure became detectable by the general dentists and lay people at 3 mm, whereas gingiva-to-lip distance was classified by these groups as noticeably unattractive at 4 mm. CLINICAL SIGNIFICANCE: The results of this study show that orthodontists, general dentists, and lay people detect specific dental esthetic discrepancies at varying levels of deviation, which may aid the dental professional in making specific treatment recommendations. PMID- 10825868 TI - Development of noncarious cervical notch lesions in vitro. AB - PURPOSE: The initiation and progression of noncarious cervical notch lesions (NCCL) continues to perplex clinicians worldwide and poses a considerable restorative challenge. The purpose of this brief communication is to report what is believed to be the first in vitro production of notch-shaped lesions in the cervical third of premolar teeth. MATERIALS AND METHODS: The lesions, were produced by axial loading of selected permanent premolar teeth in a 10% aqueous solution of sulfuric acid over a period of 5 days, followed by immersion in water for 7 days. RESULTS: Results revealed macroscopic and microscopic features similar to those observed in noncarious cervical lesions in vivo. The lesions were incidental findings while the authors were studying stress corrosion of enamel at low pH. Although much remains to be investigated regarding the etiology and pathogenesis of NCCL, axial loading and a corrosive environment may be implicated in these processes. The artificial lesions arose in clinically sound teeth, suggesting that there is no simple clinical examination to identify teeth at risk from NCCL. CLINICAL SIGNIFICANCE: The relationship between the development of NCCL and applied stress indicates that occlusal factors may play the most significant role in the initiation and progression of NCCL. PMID- 10825867 TI - Clinical study of tooth shade lightening from dentist-supervised, patient-applied treatment with two 10% carbamide peroxide gels. AB - PURPOSE: Prescribed, patient-applied tooth lightening agents, or nightguard vital bleaching, typically utilizes a 10% carbamide peroxide agent applied during nocturnal hours. The purpose of this randomized double-blind study was to compare the amount of tooth color change in two groups of subjects using dentist supervised, patient-applied 10% carbamide peroxide gel. MATERIALS AND METHODS: One group used Opalescence (Ultradent Products Inc., South Jordan, Utah) and the other NiteWhite Excel (Discus Dental, Inc., Los Angeles, California). Evaluation of tooth color for the six maxillary anterior teeth was done using a Vita shade guide at baseline, 1, 2, and 4 weeks. Subjects were instructed to apply the gel nocturnally using a custom-made soft tray 8 hours per day for 2 weeks. The 16 tabs of the shade guide were ranked according to value from darkest to lightest. The number (1-16) that correlated to the shade tab selected as the match for each tooth was the outcome variable. A Kruskal-Wallis one way analysis of variance on ranks was used. RESULTS: The test revealed no statistically significant difference between Opalescence and NiteWhite Excel for lightening the teeth (p = .807). The color change was still significant after 2 weeks without further bleaching activity. The baseline evaluation of the maxillary incisors and canines for all subjects, regardless of group, demonstrated a significant shade difference, with the canines being darker. This difference was not seen after 2 weeks of active bleaching or at the 4-week evaluation. CLINICAL SIGNIFICANCE: In this study comparing bleaching products, patients using Opalescence and NiteWhite Excel experienced a significant change in the color of their teeth relative to baseline values after 2 weeks of active treatment. PMID- 10825869 TI - Integrated orthodontic and restorative procedures for replacement of lost central incisors. AB - The use of adhesive material in association with many clinical procedures has contributed a great deal to solving functional and esthetic problems. Before adhesive techniques were known, dental procedures were more invasive and aggressive, especially in young patients. This article describes some integrated clinical procedures used to esthetically and functionally restore a mouth with traumatic loss of both central incisors. The treatment involved the orthodontic mesialization of the upper teeth toward the midline followed by anatomic and functional recovery through restorative dentistry. CLINICAL SIGNIFICANCE: This article describes some integrated clinical procedures that may effectively and conservatively improve the esthetic and functional results when a young patient loses the central incisors. PMID- 10825870 TI - Seventeen-year clinical study of ultraviolet-cured posterior composite Class I and II restorations. AB - OBJECTIVES: To compare the clinical performance of four commercial ultraviolet light-cured composite materials, and to evaluate curing-system effects on long term wear resistance of Class I and II restorations. MATERIALS AND METHODS: Approximately 32 samples of each of four different ultraviolet light-cured composites (n = 130) were inserted into conventional Class I and II cavity preparations by two clinicians. Cavosurface margins of the preparations were not beveled. Enamel walls of the preparation were etched, and the respective bonding agent was applied. Each restoration was evaluated by two clinicians at 5, 10, and 17 years. Direct evaluations were performed using modified United States Public Health Service (USPHS) criteria. Indirect evaluations were performed using the Leinfelder cast evaluation method. RESULTS: After 17 years, 65% of the restorations were recalled and pooled direct evaluations were conducted for color matching (94% alfa), marginal discoloration (100% alfa), marginal integrity (100% alfa), secondary caries (92% alfa), surface texture (72% alfa), and anatomic form (22% alfa). Mean occlusal wear from indirect evaluations at 5, 10, and 17 years was 197 +/- 85 microns, 235 +/- 72 microns, and 264 +/- 80 microns, respectively. For direct and indirect evaluations there were significant differences (p < or = .05) between the baseline and 5-year recall evaluations. CLINICAL SIGNIFICANCE: This study demonstrated that the mean pooled occlusal wear of four ultraviolet light-cured posterior composites at 17 years was 264 microns (approximately 0.25 mm), and that most wear (75%) occurred in the first 5 years. Of all recalled restorations, 76% were judged clinically acceptable at 17 years, and 22% of those exhibited no clinically detectable wear. PMID- 10825871 TI - Effects of air abrasion and acid etching on the microleakage of preventive Class I resin restorations: an in vitro study. AB - PURPOSE: To evaluate the effects of air abrasion, acid etching, and the combination of both procedures on the microleakage of preventive Class I resin restorations. MATERIALS AND METHODS: Eighty-four extracted human molar teeth were randomly assigned to seven groups with 12 teeth each. Occlusal fissures were opened with a diamond bur and etched with phosphoric acid (groups I and VI); prepared with a diamond bur without etching (group VII); air abraded with the KCP 1000, using 50-micron aluminum oxide particles without etching (group II), and with phosphoric acid etching (group III); or air abraded with 27-micron aluminum oxide particles without etching (group IV), and with phosphoric acid etching (group V). Preparations were filled with a low-viscosity resin composite (Liquicoat, groups I to V) or with a low-viscosity polyacid-modified resin composite (PrimaFlow, groups VI and VII). Six teeth in each group were thermocycled (5 degrees-55 degrees C, 2500 cycles). Dye penetration (methylene blue) was evaluated in ordinal scale. RESULTS: The number of non-thermocycled and thermocycled specimens revealing no microleakage was as follows: group I, 6 and 1; group II, 2 and 2; group III, 4 and 3; group IV, 0 and 0; group V, 1 and 0; group VI, 3 and 2; and group VII, 0 and 0, respectively. No significant differences existed between the thermocycled specimens and non-thermocycled specimens, except within group I. CLINICAL SIGNIFICANCE: Results indicated that (1) air-abrasion treatment with 27-micron particles is less effective in preventing microleakage compared to 50-micron particles, (2) air abrasion should be combined with acid etching to reduce microleakage of preventive Class I resin restorations, and (3) acid etching significantly reduces microleakage of the low viscosity polyacid-modified resin composite in preventive Class I restorations. PMID- 10825872 TI - Bilateral dental fusion of the upper central incisors: a multidisciplinary approach. AB - The multidisciplinary treatment of a case of bilateral dental fusion of the upper central incisors with maxillary constriction, crowding, and palatal inversion of one of the lateral incisors is reported. The rarity with which this anomaly appears, along with its complex characteristics, often make it extremely difficult to treat. It requires the involvement of different specialists to find the most suitable treatment for each individual patient. CLINICAL SIGNIFICANCE: This particular case of bilateral dental fusion required the involvement of orthodontists, periodontists, endodontists, and restorative specialists. For the individual evaluation of these extremely complex cases, the esthetic and functional results to be reached must be assessed compared to treatment times and biologic and economical costs. The long-term prognoses of these complex therapies also must be borne in mind. PMID- 10825873 TI - Symbiosis of esthetics and occlusion: thoughts and opinions of a master of esthetic dentistry. PMID- 10825874 TI - Esthetic smile analysis of maxillary anterior tooth width: the golden percentage. AB - With increasing application of cosmetic dental treatment comes the need for a greater understanding of esthetic principles. Scientific analysis of beautiful smiles has revealed repeatable, objective principles that can be systematically applied to evaluate and improve dental esthetics in predictable ways. Symmetry across the midline, anterior or central dominance, and regressive proportion are three composition elements required to create utility and esthetics in a smile. The Golden Proportion has been suggested as one possible mathematic analysis tool for assessing dominance and proportion in the frontal view of the arrangement of maxillary teeth. It has proven to be controversial in developing esthetically beautiful smiles and cumbersome for evaluating symmetry. CLINICAL SIGNIFICANCE: This article considers a bilateral analysis of apparent individual tooth width as a percentage of the total apparent width of the anterior segment and proposes the concept of the Golden Percentage as a more useful application in diagnosing and developing symmetry, dominance, and proportion for esthetically pleasing smiles. PMID- 10825875 TI - Effect of two abrasive systems on resin bonding to laboratory-processed indirect resin composite restorations. AB - PURPOSE: This study compared two methods of surface roughening or preparation, with or without the use of proprietary surface wetting agents, to evaluate their effect on resin cement adhesion to the following laboratory-processed, indirect restorations: Artglass (AG), belleGlass HP (BG), Concept (C), and Targis (T). Methods of surface roughening or preparation included microetching with aluminum oxide (AO): 50 microns at 34 psi and silanized silica coating, CoJet-Sand (CJ): 30 microns at 34 psi. Artglass and Concept were tested with and without the use of their respective surface wetting agents: Artglass Liquid (AGL) and Special Bond II (SB). MATERIALS AND METHODS: One hundred twenty specimens, each consisting of a pair of cylinders (7.0 x 3 mm and 4.3 x 3 mm) were fabricated. The larger cylinder or base was embedded in self-curing resin in a phenolic ring, and bonding surfaces were finished with 320-grit silicon carbide paper. Specimen pairs for each restorative material were randomly assigned to treatment groups (n = 10) and received the following surface treatments prior to cementation: group 1 (AG/AO/+AGL), group 2 (AG/AO/-AGL), group 3 (AG/CJ/+AGL), group 4 (AG/CJ/-AGL), group 5 (BG/AO), group 6 (BG/CJ), group 7 (C/AO/+SB), group 8 (C/AO/-SB), group 9 (C/CJ/+SB), group 10 (C/CJ/-SB), group 11 (T/AO), and group 12 (T/CJ). Specimen pairs were cemented with a dual-cure resin cement (Dual) and a standardized force of 1 MPa. Specimens were light-cured 40 seconds per side (80 s total), then thermocycled 300 times at between 5 degrees and 55 degrees C. Shear bond strengths (MPa) were determined using a Zwick Materials Testing Machine at a crosshead speed of 5 mm per minute. RESULTS: One-way analysis of variance (ANOVA) and Duncan's multiple range test (alpha = 0.05) by restoration type indicated no significant differences in shear bond strength between BG group 5 (29.8 +/- 5.8), BG group 6 (28.3 +/- 4.3), T group 11 (29.3 +/- 4.9), and T group 12 (29.0 +/- 4.4). Shear bond strength in AG group 3 (35.9 +/- 3.4) was significantly higher than in AG group 4 (32.4 +/- 4.0) and equal to that in AG group 2 (31.9 +/- 3.9) and AG group 1 (30.0 +/- 3.6). Shear bond strength in C group 10 (24.8 +/- 5.7) was equal to that in C group 9 (21.5 +/- 2.9), but was higher than in C groups 7 (19.4 +/- 3.1) and 8 (19.3 +/- 3.4). CLINICAL SIGNIFICANCE: Under the conditions of this study, the combination of CoJet-Sand and Artglass Liquid resulted in the highest bond strength for Artglass restorations. Microetching with CoJet-Sand or aluminum oxide followed by wetting with an unfilled adhesive was an effective surface pretreatment for dual-cure resin cementation of the four proprietary indirect resin-ceromer restorations tested. PMID- 10825876 TI - Novel approach to measure composite conversion kinetics during exposure with stepped or continuous light-curing. AB - PURPOSE: The objective of this research was to evaluate a novel approach to monitor the polymerization reaction during a light-curing exposure by using infrared (IR) spectroscopy. MATERIALS AND METHODS: An IR spectrometer was equipped to use an attenuated total reflectance (ATR) element as an IR-active substrate. The uncured composite (Herculite XRV, Shade A2, Kerr, Orange, California) was placed against the crystal, and the IR spectrum was continuously obtained during various exposure scenarios. The degree of conversion and the maximum rate of reaction were monitored at 0 mm (top surface), 1 mm, 2 mm, and 3 mm beneath the surface. The exposure conditions included continuous 40-second or 60-second exposures at 100% intensity (800 mW/cm2) or a stepped output of 10 seconds at 17% maximal output (133 mW/cm2) followed by full output for the remainder of the 40-second or 60-second exposure (Elipar Highlight, ESPE, Norristown Pennsylvania). The results were analyzed using MANOVA with appropriate post hoc tests (p < or = .05). RESULTS: For 40-second exposures, the peak conversion rates were significantly reduced (p < .05) when using the stepped exposure mode compared to the continuous exposure: 40-second top surface: stepped = 5.5%/s +/- 0.4, continuous = 10.5%/s +/- 1.0; 1 mm step = 3.6%/s +/- 0.4, continuous mode = 4.8%/s +/- 0.2. The same trend was noted when using the 60 second exposure. Equivalent conversion values (p > .05) beneath the surface between stepped and continuous exposure modes at similar depths 60 seconds after light initiation were only attained at 3 mm 4 for the 40-second exposure. However, using the 60-second exposure, equivalent conversion values between step and continuous exposure modes at similar depths were obtained. Even with a reduced conversion rate at the surface using the stepped cure mode, polymerization shrinkage forces were sufficient to debond the specimens from the test crystal after only 20 seconds into the exposure. This result indicated that stress development in the curing composite was non-uniform, and stress values developed at the surface of the restoration were the greatest. CLINICAL SIGNIFICANCE: Stepped intensity curing for the ESPE Highlight unit was shown to produce significantly lower conversion rates at the surface and at 1-mm depths, but longer exposure times were still required to provide conversion values equivalent to continuous exposure. PMID- 10825877 TI - Margin gap size of ceramic inlays using second-generation CAD/CAM equipment. AB - OBJECTIVE: The goal of this study was to determine if the fit of second generation computer-assisted design/computer-assisted manufacturing (CAD/CAM) (CEREC 2, Sirona Dental Systems, Bensheim, Germany) inlays within Class II cavity preparations were within the range of 50 to 100 microns and were equal or better to CEREC 1 inlays. MATERIALS AND METHODS: Extracted human molars free of surface defects and caries were prepared with standard mesio-occlusodistal cavity preparations. Ceramic inlays were fabricated with CEREC 2 CAD/CAM equipment and seated into cavity preparations. Digital images were captured at 100 times magnification. Marginal gaps were measured with an image analysis program. For each restoration, gaps were measured at 12 locations along interproximal margins and 10 locations along occlusal margins. RESULTS: No difficulty occurred in seating any of the milled inlays. Average marginal gaps for occlusal (59 +/- 35 microns) and interproximal (97 +/- 66 microns) margins were statistically different (t-test, p < or = .01). Average marginal gap for all sites combined was 80 +/- 57 microns. Marginal gap widths were smaller than those obtained from a similar study done earlier with the CEREC 1 unit for occlusal (89 +/- 65 microns) and interproximal (105 +/- 81 microns) margins. Ease of seating with CEREC 2 inlays was much better than with CEREC 1. CLINICAL SIGNIFICANCE: Improvements in software and hardware allow the CEREC 2 to adapt well to standard inlay cavity preparations. The ability of the device to create inlays that seat without interference is remarkable. The improved fit and ability to create the occlusal surface in a variety of ways make CEREC 2 an attractive restorative option for all-ceramic restorations. PMID- 10825878 TI - Potential for load-induced cervical stress concentration as a function of periodontal support. AB - PURPOSE: The purpose was to determine, photoelastically, the locations of occlusal load-induced stress concentrations within a maxillary premolar as a function of degree of periodontal support. MATERIALS AND METHODS: Composite three dimensional models of a maxillary first premolar were fabricated for quasi-three dimensional photoelastic stress analysis. Individual materials were used to model enamel, dentin, periodontal ligament, and alveolar bone. Three levels of periodontal support reduction (0%, 20%, 25%) were simulated by varying the socket depth. Vertical loads of 10 lb were applied to: (1) tip of buccal cusp, (2) tip of lingual cusp, and (3) center of occlusal surface. The resulting stresses were monitored and recorded photographically in the field of a circular polariscope arrangement. RESULTS: Cuspal loading concentrated stress around the cervical region below the loaded cusp, being highest under buccal cusp loading. The location of the stress concentration shifted apically as periodontal support diminished. CLINICAL SIGNIFICANCE: Observed cervical stress concentration corresponded well to the common location of abfractions seen in clinical situations. Since the location of cervical stress concentration varied with occlusal conditions, occurrence of abfractions must be highly dependent upon each patient's occlusion. Periodontal support reduction shifted the location of highest stress away from the cementoenamel junction. Therefore, abfractions are less likely to occur on periodontally compromised teeth compared to stable teeth. PMID- 10825880 TI - Esthetic compomer restorations in posterior teeth using a new all-in-one adhesive: case presentation. AB - Compomers have continually gained in importance since their introduction in 1993, especially because of their ease of handling. The main indications are cervical lesions, Class III restorations, and restorations in the primary dentition. To extend the indications of compomers to esthetic Class I and II restorations in the permanent dentition, improvements in wear resistance and bond strength need to be accomplished. Studies have shown an improved bond strength and less marginal gap formation between compomers and enamel and dentin with acid etching of the tooth before application of conventional mild self-etching adhesives. Newly formulated, improved self-etching adhesives with a higher demineralization potential recently have been developed. The objective of this case presentation is to demonstrate step-by-step the clinical procedure for the application of a newly developed self-etching adhesive with high demineralization potential in combination with a compomer material in occlusal cavities. CLINICAL SIGNIFICANCE: Improved self-etching adhesives provide a faster and simplified application technique and allow effective conditioning and priming of enamel and dentin in one step, without waiving sufficient bond strengths. With limited indications, compomers are used in combination with these self-etching primers to restore occlusal cavities in posterior teeth. Evidence-based dentistry requires exact scientific data to evaluate whether this new trend results in benefits for patients. However, until clear evidence from controlled clinical studies supports the use of improved compomer systems in Class I and II cavities in the permanent dentition, dentists should be careful with this indication. PMID- 10825879 TI - Packable composites: overview and technical considerations. AB - OBJECTIVE: New composites, called packable or condensable composites, are being promoted as amalgam alternatives. The purposes of this review article are to identify these products, define new terminology associated with them, summarize the advertised properties for the materials, discuss the ideal properties for packable composites, review the properties of the major products, and critically evaluate the proposed handling procedures for these materials. REVIEW: The term packable is preferable to condensable for describing this new class of materials. All materials should be considered amalgam alternatives, not amalgam substitutes. The compositions and physical properties reported by manufacturers reveal that none of the materials represents a remarkable improvement over the properties of more traditional universal composites. The designs of Solitaire (Heraeus Kulzer), ALERT (Jeneric-Pentron), and SureFil (Dentsply/Caulk) are discussed in detail. The distinguishing characteristics of all packable compositions are less stickiness or stiffer viscosity than conventional composites, which allow them to be placed in a manner that somewhat resembles amalgam placement. CONCLUSIONS: Packable composites may allow more convenient placement in posterior sites and may offer some technique advantages over conventional composites. However, there is no evidence that their clinical properties are consistently better than those of conventional universal composites. CLINICAL SIGNIFICANCE: Packable composites may be selected as alternatives to amalgam or conventional universal composites, but they are not equal to or better than dental amalgam in all respects. Also, in most cases, mechanical properties of packable composites are not substantially better than those of most conventional universal composites. PMID- 10825881 TI - In vitro penetration of the pulp chamber by three brands of carbamide peroxide. AB - PURPOSE: Vital tooth bleaching has become a popular procedure for whitening teeth. Most home bleaching products contain 10% carbamide peroxide. The purpose of this in vitro study was to measure the quantity of hydrogen peroxide that reaches the pulp chamber from three carbamide peroxide products: Opalescence, Sparkle, and Rembrandt. MATERIALS AND METHODS: Seventy roots of extracted premolars were amputated approximately 3 mm apical to the cementoenamel junction, and the pulp tissues were removed. They were divided into three experimental groups (n = 20) and a control group of 10 teeth. An acetate buffer solution was placed in the pulp chamber before the crown was exposed to the bleaching agent at 37 degrees C for 25 minutes. The buffer solution was removed and reacted with leukocrystal violet and horseradish peroxidase. The optical density of blue color that developed was measured at a wavelength of 596 nm and read from a standard curve for hydrogen peroxide quantity. RESULTS: The measured amounts of hydrogen peroxide were 3.605 +/- 1.405, 1.282 +/- 0.762, and 0.339 +/- 0.251 micrograms for the Opalescence, Sparkle, and Rembrandt groups, respectively. A statistically significant difference in the hydrogen peroxide levels was observed by analysis of variance (p < .05) among the three groups. It was concluded that the penetration of commercial bleaching products was different even though the products were labeled as having the same 10% carbamide peroxide. CLINICAL SIGNIFICANCE: Carbamide peroxide penetration to the pulp varies significantly for various commercial bleaching products. This may result in different levels of tooth sensitivity or bleaching efficacy. PMID- 10825882 TI - Nightguard vital bleaching of tetracycline-stained teeth: 54 months post treatment. AB - PURPOSE: The purpose of this longitudinal whitening study was to determine the stability, post-treatment side effects, and patient satisfaction after 6 months of active treatment of tetracycline-stained teeth with 10% carbamide peroxide at 0 and 54 months post treatment. MATERIALS AND METHODS: Twelve patients who completed the study (80%) were contacted and asked to participate in a survey concerning their whitening experience. Subjects were asked whether there had been any change in the shade of their teeth after treatment, and if they had experienced any side effects that they believed were treatment-related. Eight of the twelve patients underwent clinical examination. RESULTS: Ten patients (83%) reported no obvious shade change or only a slight darkening not noticed by others. Two (17%) reported a slight darkening that is probably noticeable by other people, but no one reported moderate darkening or significant darkening back to original shade. All respondents (n = 12) denied having to have a crown or root canal that they believed was treatment-related. Examiners who compared preoperative and post-treatment photographs and Vita shade values were in agreement with the patient's perceptions of shade change. The degree of improvement was significant for both the immediate (0 mo) and the 54-month post treatment comparison with the pretreatment shade (p < .005 and p < .01 respectively). CLINICAL SIGNIFICANCE: Results of this nightguard vital bleaching study indicate that tetracycline-stained teeth can be whitened successfully using extended treatment time, and that shade stability may last at least 54 months after treatment. Patients who participated in this study were overwhelmingly positive about the procedure in terms of shade retention and lack of post treatment side effects. PMID- 10825883 TI - Elective dentistry. PMID- 10825884 TI - Developing confidence and competence in cosmetic dentistry. Part III--Creating the plan: the art and science of cosmetics. PMID- 10825885 TI - A review of condensable composite. PMID- 10825886 TI - Developing confidence and competence in cosmetic dentistry. AB - When it comes to cosmetic or esthetic dentistry the old adage "beauty is in the eye of the beholder" should be a guiding principal when developing an outline of treatment for our patients. Frequently our patients come to us with requests for improving their smiles but uncertain as to exactly what they want or even worse that have expectations that we are unable to deliver. The path traveled from initial interviewing of the patient to completing each case can be fraught with pitfalls of disappointment unless a careful process of educating the patient and constant communication is followed. PMID- 10825887 TI - Differential diagnosis in dentistry. PMID- 10825888 TI - Developing confidence and competence in cosmetic dentistry. Part II of III- Treatment planning and consultation. PMID- 10825889 TI - Nightguard vital bleaching: a review of the literature 1994-1999. PMID- 10825890 TI - Dentistry's role in the management of obstructive sleep apnea. PMID- 10825891 TI - COX-2 inhibitors and dental pain control. AB - Celecoxib (CELEBREX) and rofecoxib (VIOXX) appear to offer the following advantages: reduced incidence of gastric ulceration during long-term administration; little or no effect on platelet aggregation; longer clinical duration of action than aspirin, acetaminophen and ibuprofen. However, in the context of the management of dental pain and inflammation, the following points and disadvantages should be considered: no greater effectiveness than conventional NSAIDs (e.g., ibuprofen) for dental pain; greater cost than conventional NSAIDs (especially those available in generic forms); not available over-the-counter; possible inadequate duration of action for postoperative dental pain (see references 6 and 7); similar contraindications and drug interactions to less expensive, equally effective conventional non-selective NSAIDs. At this time, celebrex and rofecoxib cannot be recommended over conventional, non selective NSAIDs as first-choice drugs for pain and inflammation in dentistry. Practitioners are cautioned against selecting any new drug based on "clinical trials of one", in which both the dentist and the patient know the drug being prescribed, (as opposed to double-blind studies), usually in the context of considerable "hype" about the drug (based on comments about the fact that the agent being tried is "new") and strong placebo reinforcement based on the dentist's enthusiasm for the new product, which does not usually accompany the prescription of older, routinely prescribed drugs. Finally, such "clinical trials of one" invariably involve close follow-up about the outcome of the treatment, which is usually not done with more common, older drugs, and which only introduces further bias into the interpretation of the effectiveness of the drug by both the patient and the dentist. Anaglesic drugs should be selected on the basis of controlled, double-blind, randomized clinical trials which utilize a reasonable, dentally-related pain model. The older NSAIDs, such as ibuprofen, naproxen, diflunisal and others, remain first-choice drugs for the treatment of mild-to-moderate pain in dentistry in patients lacking the contraindications for such drugs. As proposed by this author several years ago, the major contraindications to NSAIDs can be remembered by the "SAAB Rule", an acronym which stands for "Stomach problems", Aspirin Allergy" and "Bleeding problems", in addition to pregnancy and hepatic/renal disease. PMID- 10825892 TI - Case study. Drug-related mucosal reaction. PMID- 10825893 TI - Out of the mouths of babes. Are you spotting communication disorders in children? PMID- 10825894 TI - Case reports: lasers in soft-tissue management. PMID- 10825895 TI - Case study. Cherubism. PMID- 10825896 TI - Putting the muscle into X-rays. PMID- 10825897 TI - The hygienist's role in halitosis treatment. PMID- 10825898 TI - Case 9. Actinic cheilosis. PMID- 10825899 TI - Resuming your career. PMID- 10825900 TI - Wearing gloves: is it protection or punishment? PMID- 10825901 TI - Case study. Ameloblastic fibro-odontoma. PMID- 10825902 TI - The elderly are rich in diversity, poor in oral care. Interview by Cathleen Terhune Alty, RDH. PMID- 10825903 TI - Communication errors--are you guilty? PMID- 10825904 TI - Does ginseng boost the quality of life? PMID- 10825905 TI - Case 11. Angioedema. PMID- 10825906 TI - What happened in Kansas could happen in your state. Are you ready? PMID- 10825907 TI - Safety check. PMID- 10825908 TI - 'Cleaner' sources of water may be depriving dental patients of necessary fluoride content. PMID- 10825909 TI - Case 6. Florid cemento-osseous dysplasia. PMID- 10825910 TI - Getting paid what you're worth. PMID- 10825911 TI - Answer line spells out seven ways to ask for a raise. PMID- 10825912 TI - Following the paper trail. PMID- 10825913 TI - Massage helps relieve aching muscles, so why not invite a therapist to the office? PMID- 10825914 TI - Case 10. Actinic keratosis. PMID- 10825915 TI - A state of mind. PMID- 10825916 TI - The who's who of implant candidates. PMID- 10825917 TI - Risk management--plot out a strategy for all the 'what ifs'. PMID- 10825918 TI - Viagra presents special concerns for the hygienist and the dental team. PMID- 10825919 TI - Treatment of Lyme disease. PMID- 10825920 TI - New drugs for allergic conjunctivitis. PMID- 10825921 TI - No acute varicella-zoster virus replication in peripheral blood mononuclear cells during postherpetic neuralgia. AB - Patients suffering from postherpetic neuralgia (PHN) were investigated whether varicella-zoster virus (VZV) DNA or RNA could be detected in their peripheral blood mononuclear cells (PBMCs). Altogether 16 samples were tested by standard polymerase chain reaction (PCR) for open reading frame (ORF) 14 and ORF 29, standard and nested PCR for ORF 63, and isothermal transcription-based nucleic acid amplification (NASBA) for ORF 63 and ORF 68. By these methods neither VZV DNA nor VZV RNA could be detected. The obtained results are in contrast to those of other authors (Vafai et al., 1988; Mahalingam et al., 1995) but support the hypothesis of Bennett (1994) and Kost and Straus (1996) proposing that PHN is not caused by acute VZV replication but a consequence of neuronal damage accompanying replication of VZV in ganglia during zoster episodes. PMID- 10825922 TI - A ribozyme targeted to RNA polymerase gene of infectious bursal disease virus effectively cleaves and inhibits expression of the viral gene product. AB - Five hammerhead-type ribozymes were designed and cloned to cleave infectious bursal disease virus (IBDV) RNA and inhibit protein synthesis from cloned full length viral cDNA genes. Two ribozymes (R1 and R2) were directed to the large viral RNA segment gene and three ribozymes (R3, R4, and R5) were directed to the small viral RNA segment gene. Targets for the ribozymes were produced from cloned full-length coding regions of both small and large viral RNA segment genes. Ribozymes and their corresponding targets were synthesized as in vitro transcripts. Despite several attempts at different temperatures, no cleavage of viral RNA transcripts with four of the ribozymes (R1, R2, R3, and R5) was observed. One of the ribozymes, R4, was effective in cleaving the viral RNA polymerase gene transcripts in a magnesium-dependent manner. Ribozyme R4 caused 82% reduction in the synthesis of the viral RNA polymerase gene product. The inhibition was specific since there was no change in the rate of synthesis of the Escherichia coli beta-galactosidase protein. The results suggest that ribozyme R4 can be used as potential anti-IBDV agent. It was also demonstrated that the hammerhead-type ribozymes can cleave sites other than conventional GUC sequence motif as the cleavage site of ribozyme R4 had GUU motif which conformed to the NUX consensus motif. PMID- 10825923 TI - Preferential banding of secondary veins in strawberry is caused by mixed virus infection. AB - Leaves of Fragaria ananassa Duch. cv. Redgauntlet with mottle and mild dwarf symptoms were grafted onto F. vesca indicator clones. The youngest leaves developed specific vein banding pattern located preferentially on secondary veins near the edge of the leaves. Electron microscopy of ultrathin sections and negatively stained purified virus preparations from symptom-bearing strawberry leaves revealed presence of different-sized isometric virions. Particles of about 50 nm and 23 nm in diameter were identified as strawberry vein banding virus (SVBV) and tobacco necrosis virus (TNV) D strain. Based on results of electron microscopy, DNA hybridization, enzyme-linked immunosorbent assay (ELISA), and DNA sequencing we propose that the anomalous "leaf edge vein banding" symptoms are caused by a mixed virus infection with SVBV and other viruses such as TNV. PMID- 10825924 TI - Effect of brefeldin A on Mayaro virus replication in Aedes albopictus and Vero cells. AB - Brefeldin A (BFA), a fungal metabolite that blocks transport of newly synthesized proteins from the endoplasmic reticulum, was found to inhibit Mayaro virus replication. At the concentration of 0.05 microgram/ml, the yield of the virus was inhibited by 94% in Aedes albopictus cells and by 99.5% in Vero cells. Treatment of A. albopictus cells with BFA did not inhibit the virus protein synthesis. However, this compound drastically reduced viral protein synthesis in Vero cells. The inhibitory effect progressively declined when BFA was added at late times post infection (p.i.). The effect of BFA on protein glycosylation is discussed. PMID- 10825925 TI - A fast, sensitive and specific method for rice dwarf virus detection by northern blot hybridization. AB - Based on stability of double-stranded (ds) RNA, a new, fast, sensitive, and specific method for detection of genomic rice dwarf virus (RDV) dsRNA by molecular hybridization was developed. In contrary to the commonly used, standard Northern blot analysis, dsRNA is denatured in the immobilized state on the blot. Therefore, risk of degradation of single-stranded (ss) RNA by ribonuclease (RNase) during sample preparation, electrophoresis and blotting is eliminated. This method overcomes disadvantage of incomplete denaturation of dsRNA in Northern blot analysis. In conclusion, the newly developed method is reliable, sensitive, very specific, and gives a low background. The entire procedure is also less time consuming; it can be completed within 2-3 days. The new method may be regarded as a modification of the standard Northern blot analysis. PMID- 10825926 TI - Unintegrated viral DNA as a marker for human immunodeficiency virus 1 infection in vivo and in vitro. AB - The unintegrated viral DNA found in human immunodeficiency virus (HIV) infection includes linear and circular forms. The circular unintegrated viral DNA (CUVD) could be of either 1-long terminal repeat (LTR) or 2-LTR form. Inverse primers from nef (upstream) and gag (downstream) gene sequences of HIV-1 genome were designed to span the LTR circle junction. CUVD was assayed in unstimulated, quiescent persistently infected cell lines 8E5, HIIIB, and GB8, as well as in peripheral blood lymphocytes (PBLs) of HIV-1-infected patients by nested PCR in a cross sectional study. CUVD in the infected cell lines (in vitro) was predominantly of 2-LTR form in 8E5 and GB8 cells, while in HIIIB cells, there was besides 1-LTR and 2-LTR an additional, intermediate form. In vivo, CUVD was predominantly of 1-LTR form. The possibility of using CUVD, an early phenomenon in the virus replication, as an additional postpenetration, preintegration marker of HIV infection is discussed. PMID- 10825927 TI - Detection of low-virulent classical swine fever virus in blood of experimentally infected animals: comparison of different methods. AB - The effectiveness of virus isolation, commercial antigen enzyme-linked immunosorbent assay (ELISA), reverse transcriptase-polymerase chain reaction (RT PCR), and flow cytometry in detection of a low-virulent classical swine fever virus (CSFV) in blood in the early period of infection was evaluated. Domestic pigs at the age of 6-8 weeks and young wild boars were inoculated with a low virulent field isolate of CSFV originating from a wild boar. This virus induced serious clinical reaction in only one pig which was naturally infected with Pasteurella multocida. Nine of 13 infected domestic pigs showed viremia. All infected weanling pigs were found viraemic by virus isolation on day 6 post infection (p.i.) but virus-free by RT-PCR. The flow cytometry was apparently not as sensitive as the virus isolation. Two young wild boars infected with the virus were viremic only for the first 2 days p.i. Virus isolation and RT-PCR were of similar sensitivity. Three different commercial antigen ELISAs used were not able to detect viral antigen in any animal. PMID- 10825928 TI - Chlamydia psittaci lipopolysaccharide. A reinvestigation of its chemical composition and structure. AB - A lipopolysaccharide (LPS) of Chlamydia psittaci PK 5082 strain associated with enzootic abortion in ewes was isolated from embryonated hen eggs-grown elementary bodies (EBs) by a phenol/water procedure. Compositional analyses revealed the presence of 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo), glucosamine (GlcN), phosphorus, and fatty acids in a molar ratio of 2.6:2.0:2.4:4.4. GlcN was the only amino sugar detected. Methylation analysis of the LPS confirmed the presence of a Kdo trisaccharide proximal to lipid A having the structure Kdo-(2-->8)-Kdo (2-->4)-Kdo, which appears to be a highly conserved region in native chlamydial LPSs. The complex fatty acid composition revealed the presence of ten different straight or branched (iso and anteiso) nonhydroxy fatty acids and thirteen 3 hydroxy fatty acids. The major nonhydroxy fatty acid was icosanoic acid and the most prominent 3-hydroxy fatty acid was 3-hydroxyicosanoic acid followed by 3 hydroxy-18-methylicosanoic acid. The 3-hydroxy fatty acids represented more than two thirds of the total fatty acid content and most of them were bound in amide linkages. In contrast, most nonhydroxy fatty acids were ester-linked. It appears that LPSs from various chlamydial species differ in fatty acid composition and distribution. PMID- 10825929 TI - Cloning and sequencing of truncated gIV glycoprotein gene of an Indian isolate of bovine herpesvirus 1. AB - Bovine herpesvirus 1 (BHV-1) has been reported from the Indian subcontinent in late 70's. In order to identify the origin of an Indian isolate of BHV-1 (IBR/H 167 VS) and its molecular relationship to known strains of BHV-1, a 680 bp region of the glycoprotein gene gIV was amplified by polymerase chain reaction (PCR), cloned and sequenced. Comparison of this sequences with the corresponding one of an European strain of BHV-1 (Cooper) revealed more than 99% nucleotide (nt) homology. We conclude that the Indian isolate under study does not differ from the Cooper strain regarding the gIV gene. PMID- 10825930 TI - Partial antigenic characterization of different potato virus Y NTN strain isolates. AB - Eight isolates of potato virus Y NTN strain (PVY-NTN) of different origin were studied by means of monoclonal antibodies (MAbs) in non-competitive and competitive enzyme-linked immunosorbent assay (ELISA), and by immunoblot analysis of the viral coat protein (CP). As the MAbs reacted with the denatured viral CP, their epitopes must be continuous. The ELISA data demonstrate that the epitopes are topologically different. The epitopes may be located on the N-terminal part of CP as showed its partial amino acid sequencing and the immunoblot analysis. PMID- 10825931 TI - Transduction of antibiotic resistance in Pseudomomas aeruginosa: relationship between lytic and transducing activity of phage isolate AP-423. AB - Isolation and propagation of a wild type phage, isolate AP-423, from an apparently lysogenic strain of Pseudomonas aeruginosa, resistant to a series of anti-pseudomonadal antibiotics, and its use for transduction of resistance determinants is described. The phage isolate AP-423 showed a phenomenon of host restriction, i.e. it was lysogenic only for some of the recipient strains tested. Its transduction capacity, both in sets of genes transduced and frequency of transduction, was different in two recipient strains of P. aeruginosa. This phage showed also some restriction in titers, to which it could be propagated, only in certain recipient strains. PMID- 10825932 TI - Monoclonal antibodies suitable for type-specific identification of herpes simplex viruses by a rapid culture assay. AB - Eight monoclonal antibodies (MAbs) to herpes simplex viruses 1 and/or 2 (HSV-1, HSV-2) were tested for their reactivity with 190 previously typed HSV-positive clinical specimens in order to determine their suitability for use as type specific reagents. Using a rapid culture assay we found that two MAbs (T51 and T96) identified HSV-1 in all the 94 specimens, previously found HSV-1-positive, but did not react with the 96 specimens, previously found HSV-2-positive. In contrast, one MAb (303) confirmed the presence of HSV-2 in all the specimens, previously found HSV-2-positive, but did not react with any of the 94 specimens, previously found HSV-1-positive. These three type-specific MAbs allow for a rapid type-specific identification of HSV in infected cultures. One type-common MAb (T111) reacted with all HSV-positive cultures. This MAb can be used as an excellent reagent for detection of HSV infection. PMID- 10825933 TI - The NS5 gene location of two turkey meningoencephalitis virus genomic sequences. AB - Two new turkey meningoencephalitis virus (TMEV) nucleotide sequences were aligned to complete sequences of genomes of the flaviviruses that were available at present in the GeneBank. It was found that the both TMEV sequences represent different NS5 locations; the sequence with Acc. No. AF098456 is located downstream of that with Acc. No. AF013377 on the TMEV NS5 gene. This finding provides further insight into the TMEV NS5 gene structure and shows that the two sequences are located on the NS5 gene separately. PMID- 10825934 TI - Disaster conference gives animal issues a "seat at the table". PMID- 10825935 TI - Rabbit calicivirus infection confirmed in Iowa rabbitry. PMID- 10825936 TI - Recommends integrated approach to diagnosis and treatment. PMID- 10825937 TI - Are laws prohibiting ownership of pit bull-type dogs legally enforceable? PMID- 10825938 TI - What is your diagnosis? Chronic fibrosing pleuritis, pleural effusion, and lobar consolidation. PMID- 10825939 TI - Theriogenology question of the month. Transmissible venereal tumor (TVT). PMID- 10825940 TI - Animal behavior case of the month. Congo African Grey parrot examined because of feather picking and self-injurious behavior. PMID- 10825941 TI - Availability of training for prepuberal gonadectomy at North American veterinary colleges. PMID- 10825942 TI - Electrocardiographic assessment of antianxiety medication in dogs and correlation with serum drug concentration. AB - OBJECTIVES: To determine effects of tricyclic antidepressants (TCA) on the ECG of dogs treated for behavioral conditions and to examine correlations between ECG findings and serum concentrations of these medications. DESIGN: Repeated-measures study. ANIMALS: 39 client-owned dogs with behavioral problems. PROCEDURE: Two groups of dogs with behavioral problems were evaluated. In group 1 (n = 20), ECG tracings were recorded before starting treatment with TCA and again after treatment for > or = 1 month. Dogs in group 2 were already on long-term maintenance amounts of antianxiety medication when ECG tracings were recorded and serum concentrations of medications were obtained. RESULTS: Significant differences were not detected for dogs in group 1 between ECG values measured before and after TCA administration. The ECG values for dogs in group 2 did not differ significantly from the mean of group-1 dogs before receiving medication or from the reference range used at our facility. Duration of the P wave had a significant positive correlation with serum concentrations of clomipramine but significant negative correlation with serum concentrations of amitriptyline. The QT interval corrected for heart rate had a significant negative correlation with serum concentrations of amitriptyline. CONCLUSIONS AND CLINICAL RELEVANCE: Amitriptyline and clomipramine administered at standard dosages apparently do not cause ECG abnormalities in healthy dogs with behavioral problems. These medications should be used cautiously in dogs with conduction abnormalities, and clinicians should periodically monitor ECG and use good clinical judgment to weigh risks and benefits of medications for the safety of each dog. PMID- 10825943 TI - Comparison of absorptive capacities of original and modified Schirmer tear test strips in dogs. AB - OBJECTIVE: To compare in vitro and in vivo absorptive capacities of modified Schirmer tear test strips with those of original strips, and to establish reference values for use with the modified strips. DESIGN: Prospective study. ANIMALS: 100 dogs. PROCEDURE: In vitro absorptive capacity was determined by immersing strips in an irrigating solution for 15 seconds and recording amount of wetting. In vivo absorptive capacity was determined by placing an original Schirmer tear test strip in 1 eye and a modified strip in the other eye of 50 dogs with normal or abnormal tear production. Time required to wet 10 mm of each strip was recorded. Measurements were repeated 30 minutes later after reversing which strip was placed in the left or right eye. Reference values (mean +/- SD) were determined by recording the time required to wet 10 mm of the modified strip in 50 healthy dogs. RESULTS: Amount of wetting was significantly less and time required to wet 10 mm was significantly greater for the modified strip, compared with the original strip. Reference values determined for the modified strip were 32 +/- 11 seconds in the right eye, 33 +/- 11 seconds in the left eye, and 32 +/- 10 seconds in both eyes. CONCLUSIONS AND CLINICAL RELEVANCE: Absorptive capacities of the original and modified Schirmer tear test strips were significantly different. Reference values determined for 1 strip should not be used for the other strip. PMID- 10825944 TI - Sedative and cardiorespiratory effects of medetomidine, medetomidine-butorphanol, and medetomidine-ketamine in dogs. AB - OBJECTIVE: To determine sedative and cardiorespiratory effects of i.m. administration of medetomidine alone and in combination with butorphanol or ketamine in dogs. DESIGN: Randomized, crossover study. ANIMALS: 6 healthy adult dogs. PROCEDURES: Dogs were given medetomidine alone (30 micrograms/kg [13.6 micrograms/lb] of body weight, i.m.), a combination of medetomidine (30 micrograms/kg, i.m.) and butorphanol (0.2 mg/kg [0.09 mg/lb], i.m.), or a combination of medetomidine (30 micrograms/kg, i.m.) and ketamine (3 mg/kg [1.36 mg/lb], i.m.). Treatments were administered in random order with a minimum of 1 week between treatments. Glycopyrrolate was given at the same time. Atipamezole (150 micrograms/kg [68 micrograms/lb], i.m.) was given 40 minutes after administration of medetomidine. RESULTS: All but 1 dog (given medetomidine alone) assumed lateral recumbency within 6 minutes after drug administration. Endotracheal intubation was significantly more difficult when dogs were given medetomidine alone than when given medetomidine and butorphanol. At all evaluation times, percentages of dogs with positive responses to tail clamping or to needle pricks in the cervical region, shoulder region, abdominal region, or hindquarters were not significantly different among drug treatments. The Paco2 was significantly higher and the arterial pH and Pao2 were significantly lower when dogs were given medetomidine and butorphanol or medetomidine and ketamine than when they were given medetomidine alone. Recovery quality following atipamezole administration was unsatisfactory in 1 dog when given medetomidine and ketamine. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that a combination of medetomidine with butorphanol or ketamine resulted in more reliable and uniform sedation in dogs than did medetomidine alone. PMID- 10825945 TI - Autologous blood collection and transfusion in cats undergoing partial craniectomy. AB - OBJECTIVE: To describe the procedure for autologous blood donation and associated complications in cats undergoing partial craniectomy for mass removal. DESIGN: Prospective case series. ANIMALS: 15 cats with intracranial mass confirmed by computed tomographic scan, no evidence of renal failure, and PCV > or = 22%. PROCEDURE: One unit (60 ml) of blood was collected and stored 7 to 17 days before surgery and transfused during the perioperative period if needed. The PCV was measured before donation, before surgery, during surgery, and after surgery to assess effect of donation on PCV before surgery and effect of transfusion on PCV after surgery. Cats were evaluated for donation complications, iatrogenic anemia, and adverse reactions associated with administration of autologous blood. RESULTS: Complications associated with phlebotomy were not detected. Fifteen cats underwent partial craniectomy 7 to 17 days after blood donation; all had histologic confirmation of meningioma by examination of tissue obtained at surgery. Eleven cats received autologous blood transfusions. None of the cats received allogeneic blood transfusions. Transfusion reactions were not observed. Subclinical iatrogenic anemia was detected in 3 cats. Two cats were considered to have received excessive transfusion, and 3 cats received inadequate transfusion. All cats undergoing partial craniectomy were discharged from the hospital and were alive > 6 months after surgery. CONCLUSIONS AND CLINICAL RELEVANCE: Autologous blood donation before surgery was considered safe for cats undergoing partial craniectomy for resection of meningioma. The only complication observed was iatrogenic anemia. The procedure contributed to blood conservation in our hospital. PMID- 10825946 TI - Mycobacterial neuritis in a cat. AB - A 2-year-old spayed female domestic shorthair cat was referred for evaluation of rapidly progressive lameness of the right hind limb, which was paralyzed. Histologic examination of biopsy specimens revealed pyogranulomatous inflammation affecting the sciatic, common peroneal, and tibial nerves, and slender, beaded, acid-fast bacilli within macrophages, nerve fibers, and degenerate axons. A diagnosis of mycobacterial neuritis was made and the cat was treated with clofazimine and enrofloxacin for extended periods. Treatment was partially effective; the goal of returning normal function to the limb was not achieved, but disease progression was halted and the cat remained a viable pet. PMID- 10825947 TI - Tracheal rupture associated with intubation in cats: 20 cases (1996-1998). AB - OBJECTIVE: To characterize clinical features of tracheal rupture associated with endotracheal intubation in cats and to evaluate the most appropriate treatment for this condition. DESIGN: Retrospective study. ANIMALS: 20 cats with a history of endotracheal intubation that subsequently developed dyspnea or subcutaneous emphysema. PROCEDURE: Medical records of cats with a presumptive diagnosis of tracheal rupture associated with intubation were reviewed. Clinical and clinicopathologic data were retrieved. RESULTS: Cats were evaluated 5 hours to 12 days after a surgical or medical procedure requiring general anesthesia with intubation had been performed. Fourteen (70%) cats were evaluated after dental prophylaxis. All cats radiographed had pneumomediastinum and subcutaneous emphysema. Eighteen of 19 cats were initially treated medically. Duration of medical treatment for cats that did not have surgery ranged from 12 to 72 hours. Cats that had surgery received medical treatment 3 to 24 hours prior to the surgical procedure. Medical treatment alone was administered to 15 cats that had moderate dyspnea, whereas surgical treatment was chosen for 4 cats that had severe dyspnea (open-mouth breathing despite treatment with oxygen) or worsening subcutaneous emphysema. Eighteen cats had improvement of clinical signs, 1 cat died after surgery, and 1 cat died before medical or surgical intervention. CONCLUSIONS AND CLINICAL RELEVANCE: Most cats with tracheal rupture associated with intubation can be treated medically. Cats with worsening clinical signs (severe dyspnea, suspected pneumothorax, or worsening subcutaneous emphysema) should have surgery performed immediately to correct the defect. PMID- 10825948 TI - Laparoscopic intra-abdominal ligation of the testicular artery following castration in a horse. AB - A 364-kg (800-lb) 1-year-old mixed-breed horse was admitted for treatment of uncontrolled bleeding after castration. Multiple attempts to ligate the testicular artery through the scrotal incisions prior to referral had been unsuccessful. Because of the owner's concerns about cost, an attempt was made to control the bleeding by applying pressure to the inguinal region and administering formalin IV. However, hemorrhage continued. A decision was made to use laparoscopy to ligate the testicular artery. The horse was anesthetized and positioned in dorsal recumbency, and a routine ventral laparoscopic approach was used. The horse recovered without further complications. Laparoscopy should be considered for ligation of the testicular artery in horses with uncontrolled bleeding after castration. PMID- 10825949 TI - Isolation of respiratory bovine coronavirus, other cytocidal viruses, and Pasteurella spp from cattle involved in two natural outbreaks of shipping fever. AB - OBJECTIVE: To identify cytocidal viruses and Pasteurella spp that could be isolated from cattle involved in 2 natural outbreaks of shipping fever. ANIMALS: 105 and 120 castrated male 4- to 8-month-old feedlot cattle involved in 1997 and 1998 outbreaks, respectively. PROCEDURES: Nasal swab specimens and blood samples were collected, and cattle were vaccinated on arrival at an order-buyer barn from 4 local auction houses. Four days later, they were transported to a feedlot, and additional nasal swab specimens and blood samples were collected. Nasal swab specimens were submitted for virus isolation and bacterial culture; blood samples were submitted for measurement of respiratory bovine coronavirus (RBCV) hemagglutinin inhibition titers. RESULTS: 93 of 105 cattle and 106 of 120 cattle developed signs of respiratory tract disease during 1997 and 1998, respectively, and RBCV was isolated from 81 and 89 sick cattle, respectively, while at the order-buyer's barn or the day after arrival at the feedlot. During the 1997 outbreak, bovine herpesvirus 1 was isolated from 2 cattle at the order-buyer's barn and from 5 cattle 7 and 14 days after arrival at the feedlot, and parainfluenza virus 3 was isolated from 4 cattle 14 days after arrival at the feedlot. During the 1998 outbreak, bovine herpesvirus 1 was isolated from 2 cattle at the order-buyer's barn and on arrival at the feedlot and from 5 cattle 7 and 14 days after arrival at the feedlot, and parainfluenza virus 3 was isolated from 1 animal the day of, and from 18 cattle 7 and 14 days after, arrival at the feedlot. Pasteurella spp was cultured from 4 and 6 cattle at the order-buyer's barn and from 92 and 72 cattle on arrival at the feedlot during the 1997 and 1998 outbreaks, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that RBCV may play a causative role in outbreaks of shipping fever in cattle. More than 80% of the sick cattle shed RBCV at the beginning of 2 outbreaks when the Pasteurella spp infection rate was low. PMID- 10825950 TI - Hemorrhagic diathesis, mesenteric hematoma, and colic associated with ingestion of sweet vernal grass in a cow. AB - A 4-year-old Holstein cow from a farm where 2 cows had recently died suddenly was referred for evaluation of acute severe colic. Right flank laparotomy revealed a large mesenteric hematoma. Within 14 layer chromatographic analyses of the moldy hay and blood from the necropsied cow and the hospitalized cow were positive for dicumarol. A diagnosis of sweet vernal poisoning was confirmed on the basis of clinical and toxicologic findings. The cow was treated with supportive therapy, blood transfusions, and vitamin K1 and recovered without complications. Because sweet vernal grass is becoming common in certain areas and the use of round bales is commonplace, practitioners should be aware of the potential for this toxicosis. PMID- 10825951 TI - Synthesis and cytotoxic activities of a new benzo[c]phenanthridine alkaloid, 7 hydroxynitidine, and some 9-oxygenated benzo[c]phenanthridine derivatives. AB - [formula: see text] A new benzo[c]phenanthridine alkaloid, 7-hydroxynitidine, was synthesized by a novel synthetic procedure. The cytotoxic activity of this compound against HeLa S3 cells was strong, but not greater than those of its mother compounds, nitidine and NK109. We also synthesized other 9-oxygenated benzo[c]phenanthridine alkaloids, 7-methoxynitidine, 9-demethylnitidine, nitidine, and fagaronine, and tested their cytotoxic activities. These results suggest that the 7-hydroxy group enhances antitumor activity and an 8- or 9 hydroxy group weakens this activity. PMID- 10825952 TI - A new cryptand: synthesis and complexation with paraquat. AB - [formula: see text] Inspired by folded, nonpseudorotaxane complexes of bis(m phenylene)-32-crown-10 systems, we synthesized a new bicyclic crown ether containing two 1,3,5-phenylene units linked by three tetra(ethyleneoxy) units. The new cryptand forms a "pseudorotaxane-like" inclusion complex with N,N' dimethyl-4,4'-bipyridinium bis(hexafluorophosphate) with association constant Ka = 6.1 x 10(4) M-1, 100-fold greater than that of an analogous simple crown ether. PMID- 10825953 TI - Design and synthesis of a 1 alpha,25-dihydroxyvitamin D3 dimer as a potential chemical inducer of vitamin D receptor dimerization. AB - [formula: see text] A dimer comprising two 1 alpha,25-dihydroxyvitamin D3 units linked by an alkyl side chain at C-11 was synthesized with a view to the simultaneous binding of two vitamin D receptor (VDR) molecules and the consequent induction of VDR dimerization. The short, convergent synthesis uses a stereoselective cuprate addition to introduce the linking side chain and a key ruthenlum olefin metathesis as the dimerization step. PMID- 10825954 TI - A new class of simplified phorbol ester analogues: synthesis and binding to PKC and eta PKC-C1B (eta PKC-CRD2). AB - [formula: see text] A unique class of simplified phorbol ester analogues is described for the first time. A highly efficient retro-annelation sequence was developed in order to remove the five-membered ring from the phorbol diterpene core, allowing access to BCD ring analogues of the phorbol esters. The binding of these analogues to protein kinase C (PKC) and the truncated peptide eta PKC-C1B (eta PKC-CRD2) is also reported. PMID- 10825955 TI - The Mukaiyama-Michael reaction of N-acyl-2,3-dihydro-4-pyridones: regio- and stereoselective synthesis of cis-2,6-disubstituted 1,2,5,6-tetrahydropyridines. AB - [formula: see text] A convenient regio- and stereoselective preparation of 1,2,5,6-tetrahydropyridines of the type 1 has been developed, starting from readily available N-acyl-2,3-dihydro-4-pyridones 2. PMID- 10825956 TI - Asymmetric synthesis of alpha-methylphosphophenylalanine derivatives using sulfinimine-derived enantiopure aziridine-2-phosphonates. AB - [formula: see text] 2-Methylaziridine-2-phosphonates were prepared from enantiopure sulfinimines and were demonstrated to be versatile synthetic intermediates for the synthesis of novel alpha-disubstituted and alpha,beta trisubstituted alpha-aminophosphonate derivatives. The first asymmetric synthesis of both enantiomers of alpha-methylphosphophenylalanine is described. PMID- 10825957 TI - Radical-based asymmetric synthesis: an iterative approach to 1, 3, 5, ... (2n + 1) polyols. AB - [formula: see text] A conceptually novel approach to 1, 3, 5, ... (2n + 1) polyols based on iterative stereo-controlled homologation of chiral hydroxyalkyl radicals is reported. Starting from alpha-keto ester precursors, the general sequence of (1) ketone reduction, (2) auxiliary attachment, (3) saponification, (4) Barton esterification, and (5) radical addition provided the two-carbon homologue in 70-80% overall yield. The simplicity and generality of this iterative strategy for 1, 3, 5, ... (2n + 1) polyol synthesis suggests an attractive alternative for the preparation of molecules containing this structural motif. PMID- 10825958 TI - Mechanistic studies on the repair of a novel DNA photolesion: the spore photoproduct. AB - [formula: see text] UV irradiation of spores results in the formation of the spore photoproduct. This novel DNA photolesion is repaired in the germinating spore in a reaction catalyzed by the spore photoproduct lyase. Model studies, using a simple bispyrimidine, suggest that this repair reaction proceeds by hydrogen abstraction from C6 of the spore photoproduct followed by beta-scission of the bond linking the two pyrimidines and back hydrogen atom transfer. PMID- 10825959 TI - Synthesis of (R)-[2-2H]isopentenyl diphosphate and determination of its enantiopurity by 2H NMR spectroscopy in a lyotropic medium. AB - [formula: see text] The synthesis of (R)-[2-2H]isopentenyl diphosphate from D mannitol 1,2:5,6-bis-acetonide in 10 steps is reported. Stereospecific incorporation of the label is achieved by a BF3-catalyzed NaCNBD3 reduction of the enantiomerically pure (S)-isopropylidene oxirane intermediate. The enantiomeric excess of the penultimate precursor [2-2H]isopentenyl tosylate (> 95% ee) was determined by 2H NMR spectroscopy in a poly-gamma-benzyl-L glutamate/CH2Cl2 liquid crystal at -50 degrees C. PMID- 10825960 TI - Biosynthesis of isoprenoids in Escherichia coli: stereochemistry of the reaction catalyzed by farnesyl diphosphate synthase. AB - [formula: see text] Farnesyl diphosphate (FPP) synthase from Escherichia coli catalyzes the condensation of isopentenyl diphosphate (IPP) and geranyl diphosphate (GPP) with selective removal of the pro-R hydrogen at C2 of IPP, the same stereochemistry observed for the pig liver, yeast, and avian enzymes. PMID- 10825961 TI - Synthesis of the proposed structure of fudecalone, an anticoccidial drimane terpenoid. AB - [formula: see text] The proposed structure of fudecalone (1), an anticoccidial drimane sesquiterpene, was synthesized as a racemate in six steps starting from a known phthalide (5). Interestingly, our synthetic 1 showed conformation 1b, while the natural one was reported as 1a, and the NMR spectral data were not identical. PMID- 10825962 TI - Amine-catalyzed addition of azide ion to alpha,beta-unsaturated carbonyl compounds. AB - [formula: see text] A new protocol for the beta-azidation of alpha,beta unsaturated carbonyl compounds is described. The method employs tertiary amines as catalysts for azide addition. The azide source is a 1:1 mixture of TMSN3 and AcOH. Tertiary amines, either in solution or bound to a solid support, are efficient catalysts for the reaction. PMID- 10825963 TI - Synthesis and fluoride-promoted wittig rearrangements of alpha-alkoxysilanes. AB - [formula: see text] Lewis acid-catalyzed reaction of allyl and benzyl trichloroacetimidates with alpha-silyl alcohols was found to be a general method for the synthesis of alpha-alkoxysilanes. Upon exposure to CsF, these alpha alkoxysilanes could be made to undergo [2,3]-Wittig rearrangement with an efficiency similar to that realized by the analogous but inherently more toxic alpha-alkoxystannanes. PMID- 10825964 TI - Methyllithium-promoted Wittig rearrangements of alpha-alkoxysilanes. AB - [formula: see text] The Wittig rearrangements of alpha-alkoxysilanes, promoted by the action of methyllithium were studied. Depending on both the substrate and reaction conditions employed, [2,3]-, [1,2]-, or [1,4]-Wittig rearrangements can be realized. These rearrangements were shown to be initiated by either Si/Li exchange or deprotonation alpha to the silane. Furthermore the sigmatropic shifts can often be followed by other synthetically useful in situ chemical events. PMID- 10825965 TI - Some allylic substituent effects in ring-closing metathesis reactions: allylic alcohol activation. AB - [formula: see text] Dienes 2a-e were used to study allylic substituent effects in the ring-closing metathesis reaction (RCM). Both the steric and electronic character of the allylic substituents were found to influence alkene reactivities. Free allylic hydroxyl groups exert a large activating effect on the RCM reaction rates. PMID- 10825966 TI - Synthesis and structure of tolyporphin A O,O-diacetate. AB - [formula: see text] The revised structure of tolyporphin A O,O-diacetate (2b) was synthesized by assembling fragments 4, 5, and 12. The synthetic substance was found to be identical to the O,O-diacetate derived from natural tolyporphin A in every respect, thus establishing the relative and absolute configurations of this natural product. PMID- 10825967 TI - Improving the stereoselectivity of bakers' yeast reductions by genetic engineering. AB - [formula: see text] The stereoselectivities of bakers' yeast catalyzed reductions of beta-keto esters can be manipulated by genetic design. Strains in which two major beta-keto ester reductases are either knocked out or overexpressed have been constructed. The former approach results in whole cell biocatalysts with reversed stereoselectivity from unmodified bakers' yeast while the latter shows useful improvements in stereoselectivity. These results indicate that the "designer yeast" approach can provide useful biocatalysts for these transformations. PMID- 10825968 TI - Disiloxane-protected 2-deoxyribonolactone as an efficient precursor to 1,2 dideoxy-1-beta-aryl-D-ribofuranoses. AB - [formula: see text] Aryl C-nucleosides are analogues of natural nucleosides where the bases have been replaced with aromatic moieties. Work herein describes the highly stereoselective syntheses of non-hydrogen-bonding carbocyclic derivatives using a disiloxane-protected 2-deoxy-D-ribono-1,4-lactone as a stable and readily accessible starting material. Unlike the bis(TBDMS)-protected congener, this compound enables the use of sterically congested ortho-substituted aryllithium reagents in the initial addition reaction. PMID- 10825969 TI - Thermolysis of vic-dihydroxybacteriochlorins: a new approach for the synthesis of chlorin-chlorin and chlorin-porphyrin dimers. AB - [formula: see text] A novel approach for the preparation of symmetrical (chlorin chlorin), and unsymmetrical (chlorin-porphyrin) dimers joined with carbon-carbon linkages as models to study the "intramolecular" charge transfer is discussed. PMID- 10825970 TI - Synthesis and biological activity of cyclic peptide inhibitors of ribonucleotide reductase. AB - [formula: see text] A series of lactam-bridged peptide inhibitors (2-6) of mammalian ribonucleotide reductase (mRR) has been designed and synthesized on the basis of the heptapeptide N-AcFTLDADF (1), corresponding to the C-terminus of the R2 subunit of mRR. Inhibition studies revealed a direct relation between ring size and activity, with peptide 5 being 2.5 times more potent than peptide 1. PMID- 10825971 TI - Building fluorescent sensors by template polymerization: the preparation of a fluorescent sensor for D-fructose. AB - [formula: see text] The application of molecular imprinting in making fluorescent sensors has been hampered by the lack of suitable fluorescent tags, which would respond to the binding event with significant fluorescence intensity changes. We have designed and synthesized a fluorescent monomer which allows for the preparation of fluorescent sensors of cis diols using molecular imprinting methods. This monomer was used for the preparation of sensitive fluorescent sensors for D-fructose. PMID- 10825972 TI - A short synthesis of bicyclic dipeptides corresponding to Xxx-L-Pro Xxx-D-Pro having constrained trans-proline amides. AB - [formula: see text] A short synthesis that generates two isomeric bicyclic dipeptides having constrained, trans-proline amide bonds has been developed. One of these bicyclic dipeptides corresponds to an Xxx-L-Pro dipeptide (4), while the other isomer corresponds to an Xxx-D-Pro dipeptide (5). The two isomers are readily distinguished by their 1H NMR spectra. PMID- 10825973 TI - Separate deamination mechanisms for isomeric styrene oxide induced N1-adenine adducts. AB - [formula: see text] Styrene 7,8-oxide (SO) induced N1-2'-deoxyadenosine 5' phosphate (AMP) adducts deaminate to corresponding inosine derivatives. For the beta-isomer of N1-SO-AMP, the chiral alpha-carbon was found to be involved in the hydrolytic deamination, suggesting formation of an oxazolinium ring as an intermediate and that a water molecule attacks the benzylic carbon. The mechanism differs from the one suggested for the alpha-isomer of N1-SO-AMP, for which deamination occurs by direct attack of water at the 6-position of purine ring. PMID- 10825974 TI - Lower rim functionalized resorcinarenes: useful modules for supramolecular chemistry. AB - [formula: see text] A synthetic scheme for the selective functionalization of all cis (rccc) resorcinarene platform at the "lower rim" was developed. Self-folding and self-complementary cavitands were prepared for molecular recognition and self assembly, bearing functionality at remote sites. These molecules promise applications on solid support and as polymeric capsules. PMID- 10825975 TI - Enantioselective parallel synthesis using polymer-supported chiral Co(salen) complexes. AB - [formula: see text] The kinetic resolution of epoxides with phenols catalyzed by a polymer-supported Co(salen) complex is applied to the first enantioselective catalytic synthesis of parallel libraries. The corresponding 1-aryloxy-2-alcohols are obtained in high yield, purity, and enantiomeric excess. Further elaboration with diversity elements provides highly efficient access to important classes of pharmacologically active compounds. PMID- 10825976 TI - Synthesis of tedanolide and 13-deoxytedanolide. Assembly of a common C(1)-C(11) subtarget. AB - [formula: see text] In this Letter we describe a synthetic strategy and an efficient assembly of a common C(1)-C(11) subtarget, (-)-3, for (+)-tedanolide (1) and (+)-13-deoxytedanolide (2), architecturally complex marine macrolides displaying potent antitumor activity. Key elements of the synthesis include two iterations of the Evans aldol protocol to construct the C(1)-C(6) moiety and a stereocontrolled vinyl anion addition to generate the C(8,9) trisubstituted olefin incorporating stereogenicity at C(7). Alkylation with a model epoxide demonstrates that (-)-3 is a competent dithiane for further elaboration of the macrolide skeleton. PMID- 10825977 TI - Tremorgenic indole alkaloids. Studies directed toward the assembly of the A, F, and I rings of penitrem D: observation of an unexpected stereochemical outcome. AB - [formula: see text] In this Letter we demonstrate the viability of a highly stereoselective tandem Mannich cyclization-grammine fragmentation/addition cascade, critical for assembly of the A and F rings of penitrem D. We also explored simultaneous execution of this tactic with concurrent construction of ring I. Reinvestigation of a model system provided an explanation for the unanticipated stereochemical outcome at C(28). PMID- 10825978 TI - Mechanism and conditions for highly enantioselective epoxidation of alpha,beta enones using charge-accelerated catalysis by a rigid quaternary ammonium salt. AB - [formula: see text] Highly enantioselective (up to 130:1) epoxidation of a variety of alpha,beta-enones to form alpha,beta-epoxy ketones is described along with a rational analysis of the mechanistic basis for this strong absolute stereochemical control by the chiral catalyst 2. PMID- 10825979 TI - Palladium-induced cyclizations for the synthesis of cis-2,5-disubstituted-3 methylenetetrahydrofurans: studies of the C7-C22 core of amphidinolide K. AB - [formula: see text] The diastereoselective synthesis of cis-2,5-disubstituted-3 methylenetetrahydrofurans via Pd(0)-catalyzed cyclization of 2-methylene-1,4 diols is described. Investigations into the scope of the reaction and its application toward the synthesis of amphidinolide K is reported. PMID- 10825980 TI - Synthesis of (+)-casuarine. AB - [formula: see text] The first synthesis of (+)-casuarine, a pentahydroxy pyrrolizidine alkaloid, is described. The key bond-forming events occur in a tandem [4 + 2]/[3 + 2] nitroalkene cycloaddition involving nitroalkene 6, chiral vinyl ether 7b, and vinyl silane 4. This process also creates five of the six stereocenters present in this potent glycosidase inhibitor. Completion of the synthesis required only four additional steps and delivered (+)-casuarine in 20% overall yield. PMID- 10825981 TI - Total synthesis of (+/-)-isoschizogamine. AB - [formula: see text] Isoschizogamine has been prepared for the first time. The synthesis requires eight steps from a readily available ketone starting material and features an aminal-forming cyclization that is based on a proposed biosynthetic transformation. PMID- 10825982 TI - The first total synthesis of (-)-tamandarin A. AB - [formula: see text] Tamandarin A (1), a newly isolated natural product similar in structure to didemnin B (2), was shown to be somewhat more active in vitro than 2 against pancreatic carcinoma with an ED50 value 1.5 to 2 ng/mL. We report here the first total synthesis of 1. The key steps include a practical stereoselective synthesis of the Hiv-isostatine unit, high-yielding linear precursor formation, a successful macrocyclization, and coupling of the macrocycle with the side chain to afford tamandarin A (1). PMID- 10825983 TI - Models of F.H contacts relevant to the binding of fluoroaromatic inhibitors to carbonic anhydrase II. AB - [formula: see text] Complexes formed between fluorobenzene and N-methylformamide or benzene have been used as models of the interaction of fluoroaromatic drugs with carbonic anhydrase II. These structures have been investigated via ab initio and density functional methods, including HF, B3LYP, and MP2 procedures. The results of the calculations are consistent with the hypothesis, suggested originally by experimental X-ray crystal structures of the drug-receptor complexes, that favorable fluorine-hydrogen interactions affect binding affinity. PMID- 10825984 TI - Staminolactones A and B and norstaminol A: three highly oxygenated staminane-type diterpenes from Orthosiphon stamineus. AB - [formula: see text] Staminolactones A (1) and B (2) and norstaminol A (3), three highly oxygenated staminane-type diterpenes having mild cytotoxic activities against highly liver-metastatic colon 26-L5 carcinoma cells, were isolated from the aerial part of the Vietnamese medicinal plant Orthosiphon stamineus (Lamiaceae). Their structures were elucidated on the basis of the extensive spectral analyses. PMID- 10825985 TI - Synthesis of C-11 methyl-substituted benzocycloheptapyridine inhibitors of farnesyl protein transferase. AB - [formula: see text] Synthesis of C-11 methyl-substituted benzocycloheptylpyridine tricyclic compounds has been achieved via two different methods. Methylation of C 11 has been effected by treatment of amine 4 with BuLi followed by Mel quenching. In a similar procedure, introduction of a C-11 substituent with concomitant rearrangement of the exocyclic double bond has been carried out. Potent farnesyl protein transferase inhibitors have been synthesized using the above methodologies. PMID- 10825986 TI - Synthesis and DNA binding of spirocyclic model compounds related to the neocarzinostatin chromophore. AB - [formula: see text] Spirocyclic model compounds which mimic the molecular architecture of one of the decomposition products of the antitumor agent NCS chrom have been synthesized. These readily accessible molecules bind with remarkable efficiency to bulged DNA oligonucleotides, offering potential for the design of therapeutic agents. PMID- 10825987 TI - Lewis acid-mediated displacements of alkoxydioxolanes: synthesis of a 1,2 dioxolane natural product. AB - [formula: see text] Addition of electron-rich alkenes to the peroxycarbenium ions derived from Lewis acid-mediated ionization of 3-alkoxy-1,2-dioxolanes provides an efficient route for the synthesis of substituted 1,2-dioxolanes. The methodology is illustrated with a rapid synthesis of a 1,2-dioxolane natural product related to the plakinic acids. PMID- 10825988 TI - Solid-phase synthesis of pyridones and pyridopyrazines as peptidomimetic scaffolds. AB - [formula: see text] We report the syntheses of peptidomimetic opioids containing the core structure N-alkyl-2-alkyl-2,3-dihydro-4-pyridone. By employing imines bound on a solid support and the Danishefsky diene, this [4 + 2] cyclocondensation reaction facilitates the synthesis of novel complex heterocycles. The central reaction is carried out under mild conditions and employs readily available building blocks. In this study we demonstrate the suitability of N-alkyl-2-alkyl-2,3-dihydro-4-pyridones as a central scaffold for peptidomimetics and establish the scope of this [4 + 2] cyclocondensation reaction with imino acids on a solid phase. We also combine the synthesis of diketopiperazines with the [4 + 2] cyclocondensation reaction to form a 9,9a dihydro-2H-pyrido-[1,2a]- pyrazine-3,8(1,4-dialkyl)dione, a bicyclic molecule containing a pyridopyrazine core structure. PMID- 10825989 TI - Improved synthesis of epothilone B employing alkylation of an alkyne for assembly of subunits. AB - [formula: see text] A strategy for assembling the two principal modules of epothilone B was developed that merges an allylic bromide with a terminal acetylene to fabricate the C10-C11 bond of the macrocycle. The resulting alkyne was semihydrogenated to give a seco ester previously employed in our total synthesis of epothilone B. This new approach affords a more efficient route to the naturally occurring macrolide and to its 9,10-dehydro analogue. PMID- 10825990 TI - Applications of aziridinium ions. Selective syntheses of beta-aryl-alpha,beta diamino esters. AB - [formula: see text] alpha,beta-Diamino esters are readily prepared through stereospecific and regioselective opening of an aziridinium ion intermediate with a variety of amines. The aziridinium ion is generated from the epoxide in two steps. PMID- 10825991 TI - Synthesis and stereochemical assignment of exo- and endo-7-methyl-7 azabicyclo[2.2.1]heptan-2-ol. AB - [formula: see text] The syntheses of both exo and endo stereoisomers of 7-methyl 7-azabicyclo[2.2.1]heptan-2-ol were achieved in straightforward fashion. Alternatively, the intramolecular cyclization of syn-4-N-methylaminocyclohexane 1,2-epoxide was found to give exo-7-methyl-7-azabicyclo-[2.2.1]heptan-2-ol as the sole product. The stereochemistry of the exo isomer was unequivocally confirmed by X-ray crystallography. PMID- 10825992 TI - Two syntheses of the 16- and 17-membered DEF ring systems of chloropeptin and complestatin. AB - [formula: see text] Two syntheses of a model system of the DEF ring system of complestatin and chloropeptin are described. The key step in both of these syntheses involves the formation of the biaryl linkage using a palladium catalyzed Suzuki cross-coupling reaction and a catalytic enantioselective ene reaction to form the 6-bromo-D-tryptophan. Additionally, ring contraction of the 17-membered DEF ring system of complestatin generates the 16-membered DEF ring system of chloropeptin in a biomimetic fashion. PMID- 10825993 TI - An organoiron approach to the benzophenone appendage of the protein kinase C inhibitor balanol. AB - [formula: see text] A new synthetic route to the benzophenone appendage of balanol, based on sequential iron-assisted nucleophilic aromatic substitution and ring-opening as well as regioselective oxidative cyanation, is described. PMID- 10825994 TI - An efficient one-flask synthesis of N-confused tetraphenylporphyrin. AB - [formula: see text] N-Confused meso-substituted porphyrin is a porphyrin isomer previously available from one-flask porphyrin syntheses as a low-yield byproduct (< 7.5%). We have found that methanesulfonic acid catalyzed condensation of pyrrole and benzaldehyde followed by DDQ oxidation provides N-confused tetraphenylporphyrin (NC-TPP) in up to 39% yield in analytical scale experiments. Preparative synthesis provided an isolated yield of 35% (800 mg). This represents a > 5-fold yield improvement and makes significant quantities of NC-TPP readily available. PMID- 10825995 TI - Cytotoxic alkaloids motuporamines A-C: synthesis and structural verification. AB - [formula: see text] The unusual structure and biological properties of the marine alkaloids motuporamines A-C, as well as the uncertainty as to the position of the olefin within the ring of motuporamine C, led us to synthesize these compounds. The strategy utilized the ring-closing metathesis reaction to form the 14- and 15 membered rings and Michael addition and amidation chemistry to introduce the spermine-like unit. The syntheses, structure assignment verifications, and also the determination of the position of the olefin in motuporamine C are described. PMID- 10825996 TI - A method for constructing the C44-C51 side chain of altohyrtin C. AB - [formula: see text] A method to construct the C44-C51 side chain of altohyrtin C has been developed and applied to a model aldehyde derived from D-glucose. The approach relies on a Wittig reaction to couple the side chain to an aldehyde and utilizes an allylic diazene rearrangement to place the C45 double bond in the correct position. PMID- 10825997 TI - Total synthesis of (+)-thiazinotrienomycin E. AB - [formula: see text] The first total synthesis of (+)-thiazinotrienomycin E (1), member of a novel class of cytotoxic ansamycin antibiotics, has been achieved. The synthesis features a highly efficient construction of the aromatic fragment 3 incorporating TBS protection of the aniline, a significantly improved synthesis of (-)-19, an intermediate employed in our trienomycins A and F total syntheses, application of the Kocienski modified Julia protocol to elaborate the E,E,E triene subunit, an efficient union of 3 and (+)-4, and Mukaiyama macrolactamization to access the thiazinotrienomycin macrolide. PMID- 10825998 TI - A two-step total synthesis of the natural pentacycle trichodimerol, a novel inhibitor of TNF-alpha production. AB - [formula: see text] Trichodimerol (1) can be synthesized by a remarkable dimerization of the chiral hydroxy dienone 5. PMID- 10825999 TI - Viral hepatitis C. AB - Identification of a cDNA clone and the genome of hepatitis C virus in 1988-1989 allowed the development of clinical tests that are now used to detect and quantify hepatitis C virus. This has largely eliminated post-transfusion hepatitis C virus infection; however, the overall incidence of chronic hepatitis C and its complications has greatly increased because of its transmission by other means, lack of a protective vaccine, and inadequate virucidal therapy. Drug abuse is the most common cause of hepatitis C; an etiologic mechanism, however, remains unknown in one third of patients referred to the New Jersey Medical School Liver Center. Response to treatment depends on the viral subtype, immune reactivity of the host, and hepatic pathologic alterations. Many patients with hepatitis C improve or are cured by administering an interferon with or without ribavirin; patients refractory to these measures exhibit persistent elevation of serum cytokines and progressive liver disease. New measures, including protease inhibitors and adjunct immunotherapy, should increase effectiveness of therapy, diminishing hepatitis C virus-induced cirrhosis and hepatocellular cancer. Populations, including the underserved, who harbor and transmit hepatitis C virus require special assistance. This is best achieved by community support groups organized through medical schools, physician associations, and churches to help prevent, detect, and treat chronic hepatitis C. PMID- 10826000 TI - Post-extubation stridor: risk factors and outcome. AB - Post-extubation stridor is a potential complication of endotracheal intubation. The incidence, risk factors, and outcome in adult patient populations are poorly defined. It was our clinical impression that the occurrence of post-extubation stridor in our medical intensive care unit was more frequent than generally reported. We therefore monitored all intubated patients to determine the incidence of post-extubation stridor and to identify any predisposing factors. All adult patients requiring endotracheal intubation and medical intensive care admission were prospectively observed over a 5-month period in a university affiliated medical intensive care unit. Sixty-seven patients requiring intubation and medical intensive care admission were evaluated. Twenty-two patients were excluded from analysis because no extubation was attempted. The remaining 45 patients were divided into two groups: 8 with and 37 without post-extubation stridor. Of the parameters analyzed, cuff pressure, treatment with corticosteroids at the time of extubation, and the presence of a primary neurologic process necessitating intubation differed significantly between groups. Six of the 8 patients who developed post-extubation stridor required reintubation. Four of these patients were subsequently successfully extubated, one required tracheostomy, and one patient died after a 2-week deteriorating clinical course. The incidence of significant stridor in our population was 17.8%. Medical management was successful in the majority of patients with post extubation stridor. Routine tracheostomy following a single episode of post extubation stridor is not indicated. Further investigation regarding risk factors and a placebo-controlled trial evaluating the efficacy of systemic corticosteroids before extubation in individuals at risk for developing post extubation stridor are needed. PMID- 10826001 TI - Esophageal candidoma in a patient with acquired immunodeficiency syndrome. AB - Oral thrush and esophagitis caused by Candida are common in patients infected with the human immunodeficiency virus. We present the case of a 33-year-old man with acquired immunodeficiency syndrome who developed dysphagia during a hospitalization for pneumonia. Signs and symptoms were consistent with Candida esophagitis. Despite therapy with fluconazole, the patient's symptoms persisted. At upper endoscopy, a 1-cm, polypoid esophageal mass at 30 cm from the incisors and several other nodular lesions were observed; white plaques were noted throughout the esophagus. Biopsy specimens of the mass contained hyphal forms consistent with Candida species. Therapy with amphotericin B improved the patient's symptoms, and resolution of the mass was confirmed by repeat upper endoscopy. We believe this is the first case in the medical literature of a Candida mass (candidoma) causing dysphagia in a patient with acquired immunodeficiency syndrome. Candidoma should be considered in the differential diagnosis of dysphagia in patients with human immunodeficiency virus infection or immunosuppression due to other causes. PMID- 10826002 TI - Measuring success: the future of the AAMP. Affirmative opportunity, not affirmative outcome. PMID- 10826003 TI - Complementary and alternative medicine: herbal therapies for diabetes. AB - Interest in and use of complementary and alternative medicine have increased in recent years, with as much as 40% of the general population having used some form of complementary and alternative medicine in the past year. Although recent national surveys have not adequately assessed the use of complementary and alternative medicine in minority populations, its use in many minority populations is at least equal to and possibly greater than that in the general population. One condition for which minority populations are likely to use complementary and alternative medicine therapies is diabetes, which is particularly prevalent in Mexican-American and Native-American communities. Many minority cultures have a long history of using herbal preparations to treat diabetes, and recent research suggests that some herbal therapies may have a role in the treatment of this complex disease. However, many questions remain regarding the proper use of herbal therapies for diabetes, particularly in regard to dosage and contaminants. Therefore, physicians who work with minority populations should ask patients about their use of these therapies and must become knowledgeable about their safety and efficacy. PMID- 10826004 TI - Cardiovascular disease in patients with diabetes: clinical considerations. AB - Cardiovascular disease is the major cause of morbidity and mortality in patients with diabetes. Diabetic individuals have a 200% to 400% greater risk for vascular disease than nondiabetics, with a disproportionately greater burden of disease complications in non-white minorities. Although the atherosclerotic plaques in the two groups are similar, diabetics have more severe and more diffuse disease than nondiabetics. Recent advances in the treatment of coronary disease have improved survival for diabetics and nondiabetics, but diabetics still have double the case fatality rate as nondiabetics, and diabetic women have particularly poor outcomes. Diabetic individuals also have an increased frequency of silent ischemia, systolic and diastolic left ventricular dysfunction, and cardiac autonomic neuropathy. The high frequency of modifiable risk factors provides great opportunities for prevention, the cornerstones of therapy being glycemic control, aggressive risk factor modification, and ongoing patient surveillance and monitoring to facilitate early disease detection and prompt intervention. In patients with coronary disease who require revascularization, both mechanical coronary interventions and bypass surgery are effective therapies. Patients with multivessel coronary disease have better results following bypass surgery with arterial grafts than following coronary interventions. However, diabetic patients are at increased risk for poor long-term outcome following either revascularization modality, with high rates of restenosis following mechanical interventions and the development of atherosclerosis in conduits following bypass surgery. PMID- 10826005 TI - The effects of pentoxifylline on spinal cord blood flow after experimental spinal cord injury. AB - Previous clinical and experimental investigations have suggested that pentoxifylline, a methylxanthine, can improve cerebrovascular circulation and reduce cerebral edema in cerebrovascular disorders. Pentoxifylline's mechanism of action includes such rheologic effects as enhanced red cell deformability, alterations in leukocyte activation, and modification of coagulation parameters. The purpose of our investigation was to determine the effects of pentoxifylline in an experimental spinal cord injury model. A compression device was used to cause a reproducible spinal cord injury in adult female albino rats. Spinal cord blood flow was monitored using a laser Doppler flow meter pre- and postinjury for 4 hours. The experimental group (N = 7) was injected with pentoxifylline 10 minutes prior to injury. The control group (N = 5) received an identical protocol, except that this group was injected with an equal amount of saline. Results of this investigation revealed that pentoxifylline treatment significantly increased spinal cord blood flow. In the pentoxifylline-treated group, spinal cord blood flow was significantly higher from 120 to 240 minutes postinjury compared with that of the control group. We conclude that via its multiple physiologic effects, pentoxifylline significantly improves spinal cord blood flow in experimental spinal cord injury. PMID- 10826006 TI - Has the well run dry? Priming the diversity pump in PhD programs. AB - America has always described itself as a melting pot of people and cultures. Perhaps it has become more of a "boiling pot" as assaults against affirmative action and charges of racism run rampant. Underrepresented minorities in the United States remain well below the national average with regard to education, employment, and health status. While today these minorities make up nearly 25% of the US population, historically they continue to be underrepresented in the medical and research professions. Some progress has been made during the last several decades, but it has stalled. And there is reason to believe that we are facing potentially rapid reversals of what has been painfully achieved. There is a significant pipeline problem in the production of minority doctorate recipients in America today. A review of a cohort of 17-year-old white and black students using a national longitudinal survey of the high school class of 1980 determined that 28% of white students graduated from college, compared with only 11% of black students--a 60% difference. Less than 1% of this original cohort of black students went on to earn science degrees. Doctoral programs now increasingly compete for the same small group of academically qualified minority students. The only long-term solution is to improve the educational opportunities starting from the early stages of the educational pipeline, beginning in elementary school. High school is too late. This process is complicated and will be costly. Pipeline initiatives will demand creativity, flexibility, and a commitment from all of us. We must increase the motivation and preparation of minority students regarding careers in science, engineering, and health care. We in academic health centers must play a key role in strengthening science education and in changing our behavior. We need to address the reasons why more than half of black students who enter college fail to graduate, why there is so little interest in science, and why minority students are not better prepared in science while they are in secondary school. And we need to address these issues now. PMID- 10826007 TI - Emerging concepts in antihypertensive therapy: the benefits of angiotensin II blockade. AB - Essential hypertension affects more than 40 million Americans, or one in four adults. The prevalence of hypertension is greater among the African-American population, with a distressingly high rate of end-organ complications. Although diabetes mellitus has surpassed hyper tension as the dominant etiology of end stage renal disease in the United States, kidney failure secondary to hypertensive nephrosclerosis remains a significant problem, particularly among African Americans. During the past decade, a shift in the paradigm for the renin angiotensin system (RAS) has evolved from a circulating vasoactive cascade toward angiotensin II (ANG II) formation at the cellular level. The molecular components of the RAS have been identified in cells, documenting the existence of an autocrine tissue RAS, as well as the presence of enzymes, which catalyze the formation of ANG II by angiotensin-converting-enzyme-independent pathways, providing new targets for therapeutic intervention. The latter challenge has important clinical implications, in view of recent evidence implicating ANG II in pathologic cell growth and cell death and fundamental events in the remodeling of the vascular wall and myocardium in the setting of hypertension. This review focuses on ANG II as a major determinant of end-organ damage in essential hypertension; the benefits of ANG II blockade at the end-organ level, which appear to be independent of the blood pressure-lowering effect; and the emerging role for ANG II receptor antagonists as first-line agents in the treatment of essential hypertension. PMID- 10826008 TI - Epidemiologic pattern of esophageal cancer at an inner-city university hospital. AB - The incidence of esophageal adenocarcinoma has increased significantly during the past 25 years in the United States and Europe. This increase has occurred predominantly among white men. To determine the effect of ethnicity and selected clinical features on the type of esophageal cancer in an urban, minority population, we retrospectively reviewed esophageal cancer at our institution. All patients with esophageal cancer from 1980 to 1995 were identified using the tumor registry data base and patient medical records at UMDNJ-University Hospital. Inclusion criteria were self-reported ethnicity and a confirmed pathologic diagnosis of either esophageal adenocarcinoma (ADENO) or squamous cell carcinoma (SCCA). Data abstracted from the record included age and year of diagnosis, weeks of survival, and risk factors, such as Barrett's esophagus and tobacco and alcohol abuse. Of 150 cases of esophageal cancer, 139 (93%) were SCCA and 11 (7%) were ADENO; the male:female ratio was 11:4. African and Latino Americans comprised 87% and white Americans 13% of the group. The incidence of ADENO increased during the study period: 1980-1984, 1 case; 1985-1989, 3 cases; and 1990-1995, 7 cases (P = .022); whereas the incidence of SCCA remained constant during the same intervals: 51, 52, and 36 cases, respectively (P > .05). By ethnicity, ADENO occurred more frequently among whites (7/19, 37%) than among African and Latino Americans (4/131, 3%); SCCA was more common among African and Latino Americans (127/131, 97%) than among whites (12/19, 63%) (P < .001). Other risk factors did not influence the type of esophageal cancer. The study concluded that the incidence of ADENO increased, primarily among white men, from 1980 to 1995 at UMDNJ-University Hospital. In contrast, the incidence of SCCA remained constant and was the primary type of esophageal cancer in African and Latino Americans. This study supports previous reports that ethnicity influences the histology of esophageal cancer. PMID- 10826009 TI - Lead poisoning from a retained bullet: a case report and review. AB - A 47-year-old man with a prior gunshot wound presented with arthritis, constipation, abdominal pain, and weight loss. Arthrocentesis did not reveal the cause of the arthritic complaints, but lead poisoning was suspected and confirmed. We present this case along with a short review of the literature pertaining to this often overlooked and reversible cause of lead poisoning. PMID- 10826010 TI - Colorectal cancer epidemiology in minorities: a review. AB - Colorectal cancer is the second leading cause of cancer death in the United States. In 1997, more than 131,000 new cases and more than 54,000 deaths were estimated. Racial and ethnic disparities in incidence, mortality and survival rates, and trends exist for this disease. Differences in colorectal cancer screening, early detection, and treatment in minority communities are related to therapeutic outcomes. Age-adjusted incidence rates for men with colorectal cancer are highest for Alaskan native men, followed by Japanese, then African-American men. For women, the incidence is highest for Alaskan native women, followed by African-American, then Japanese women. Mortality rates in men are highest for African Americans, followed by Alaskan natives and then Hawaiians. In women, mortality rates are highest for Alaskan natives, then African Americans and whites. Colorectal cancer screening rates vary by race, income, and education. It is interesting that, when compared with whites, African-American men demonstrate the higher reported rate of screening for this disease. In addition, site specificity is different for African Americans compared with whites. Findings also reveal that stage at diagnosis is an influential factor with regard to mortality and survival. This may be related in part to socioeconomic factors, differences in anatomic site, and treatment differences in African Americans. Risk factor data for this disease are scarce for minority populations. Documented differences in colorectal cancer incidence, mortality, and survival rates exist between minorities and whites. Additional research is needed on risk factors specific to African Americans and other minorities, differences in treatment, and the role of socioeconomic status. PMID- 10826011 TI - Racial and ethnic factors in the genetic pathogenesis of colorectal cancer. AB - Colorectal cancer can develop by two distinct pathogenic mechanisms: one involving chromosomal breakage and aneuploidy (called chromosomal instability) and one involving mutations at DNA micro-satellite sequences (termed micro satellite instability). Relatively few reports consider these mechanisms of colorectal cancer development across racial or ethnic groups. Available data indicate a moderate increase in colorectal cancer risk among Ashkenazi Jews who have a mutational polymorphism at codon 1307 in the APC gene. In American blacks, there is evidence for a higher prevalence of right-sided colonic tumors and an earlier age of onset of colorectal cancer. In addition, blacks have the highest colon cancer incidence in the United States among ethnic groups and have poorer 5 year survival rates compared with whites. While some differences may be attributed to health care access and socioeconomic differences, these do not completely explain all the variances. In the chromosomal instability pathway, there are polymorphisms within the P53 gene that are more prevalent in blacks, but the significance of these polymorphisms is not fully known. Blacks are more likely to demonstrate micro-satellite instability in their tumors; however, the mechanism for this phenomenon in blacks is unexplored. Differences in diet among racial and ethnic groups and polymorphic variations in drug metabolizing or acetylation genes have not been adequately cataloged. Identification of genetic and environmental factors among racial and ethnic groups should offer some insights into the observed epidemiologic data and advance opportunities to better understand the control and development of colorectal cancer. PMID- 10826013 TI - The Baltimore Alliance for the Prevention and Control of Hypertension and Diabetes: a model for developing a colorectal cancer community outreach program. AB - Many diseases that account for the majority of morbidity and mortality in African Americans are also associated with racial disparity. Such diseases include cardiovascular disease, diabetes, hypertension, cancer, acquired immunodeficiency syndrome, and violent injuries. Epidemiologic associations find the excess burden for these diseases to be rooted in biological, psychosocial, and socioeconomic factors. Community-based efforts often function in isolation, attempting to have impact on diseases that affect local residents; unfortunately, such programs cannot be comprehensive in their approach. Foreseeable gaps allow morbidity and mortality to continue to exist. The Baltimore Alliance for the Prevention and Control of Hypertension and Diabetes (the Alliance), based at the University of Maryland School of Medicine, provides an infrastructure where successful community-based programs that target hypertension and diabetes join together in collaborative effort. The Alliance creates an association that supports community based efforts, helping improve the efficiency and efficacy of such programs. Such associations can help close gaps, enabling programs to have a more comprehensive impact on communities. PMID- 10826012 TI - Role of diet and NSAIDs in the chemoprevention of colorectal cancer. AB - Colorectal cancer is a major public health problem. It is also one of the most preventable cancers. Although the colorectal cancer incidence and mortality in whites have been declining over the past 2 decades, these statistics are rising in nearly all ethnic minority groups. The development of colorectal cancer is influenced by exogenous factors, such as dietary constituents and drugs. While reputable data on the chemopreventive effects of diet and drugs, such as nonsteroidal anti-inflammatory drugs (NSAIDs), in specific minority groups are limited, evidence suggests that dietary factors may affect colorectal carcinogenesis in various ethnic minority groups. Clearly, more studies are necessary to resolve these questions. Because the risk of colorectal cancer is increasing in minority patients, they cannot wait for the results of such studies. Therefore, until definitive data are available, it is prudent for physicians to recommend that all individuals be screened for colorectal cancer according to accepted guidelines and to educate them regarding healthful eating habits that will prevent the development of colorectal cancer. Physicians should be particularly vigilant in recommending these approaches to minority patients. Patients should be advised to consume a well-balanced, low-fat, high-fiber diet that is rich in fruits and vegetables. PMID- 10826014 TI - Death postponement and increased chronic lung disease: the hidden costs of mortality reduction in the post-surfactant era. AB - To determine changes in the morbidity and mortality of ventilated, very low-birth weight infants before and after the introduction of surfactant therapy in 1990, we retrospectively studied the infants in a tertiary neonatal intensive care unit. All ventilated patients admitted from January 1, 1984, to December 31, 1997, with birth weight of 1500 g or less, excluding 76 transferred to other hospitals, were included in the analysis (N = 1336). The primary outcomes studied included mortality, length of stay, and survival without bronchopulmonary dysplasia. Although post-surfactant mortality and pulmonary air leak for infants with birth weight greater than 750 g decreased more than 50% (P < or = .026), the proportion of infants surviving without bronchopulmonary dysplasia also decreased (P < or = .034). For surviving infants with birth weight of 751 to 1000 g, mean post-surfactant length of stay increased 16 days (P = .008). Postnatal age at death also increased in the post-surfactant period for infants with birth weight of 750 g or less (P = .002). For infants with birth weight of 1000 g or less, post-surfactant increases were seen in the mean duration of assisted ventilation (+22% to 32%, P < or = .005) and the incidence of bronchopulmonary dysplasia at 36 weeks' postmenstrual age (+62% to 162%, P < .001). For all infants, the incidence of bacteremia, duration of supplemental oxygen therapy, and likelihood of discharge on home oxygen were increased in the post-surfactant period (P < or = .011). The implicit benefits of mortality reduction in the post-surfactant era may be offset by increasing respiratory morbidity in some survivors and by an unwelcome prolongation of death for some nonsurvivors. We speculate that the ultimate costs of these undesirable outcomes may greatly surpass the ostensible cost benefits of neonatal surfactant therapy. PMID- 10826015 TI - Immunoproliferative small intestinal disease: case report and literature review. AB - Immunoproliferative small intestinal disease (IPSID) is a subtype of lymphoma of mucosa-associated lymphoid tissue. Notable for a high production of alpha-heavy chains, it is designated alpha-heavy-chain disease. IPSID is a debilitating disease that has a predilection for impoverished populations of developing countries. It has been documented primarily in subjects of Middle Eastern countries and thus was previously referred to as Mediterranean lymphoma. We report the case of a 42-year-old man from Senegal who presented with chronic diarrhea, dehydration, and weight loss. The endoscopic, pathologic, and serologic findings before, during, and after treatment with fludarabine phosphate are presented. We review the literature concerning current concepts on the etiology, pathogenesis, and management of IPSID. PMID- 10826016 TI - Esophageal cancer: an unusual endoscopic appearance and presentation. AB - Esophageal cancer in advanced stages grows to occlude the esophageal lumen; presenting symptoms include dysphagia and weight loss. Esophageal cancer rarely grows to occupy a narrow column of the esophagus or manifests neurologic symptoms. We report the case of a 58-year-old man with a history of tobacco and alcohol abuse and chronic obstructive airway disease who presented with headaches, left-sided weakness, unsteady gait, and weight loss. Physical examination showed left-sided weakness. Computed tomographic scan of the brain and chest revealed, respectively, a right frontoparietal mass and a tumor mass in the distal esophagus. The patient's weakness and headaches improved after treatment with dexamethasone and craniotomy with partial enucleation of the brain lesion. An esophagogastroduodenoscopy revealed a large, elongated mass in the esophagus. Pathologic analyses of biopsies of the esophageal mass showed mixed adenosquamous carcinoma. The brain mass histology showed poorly differentiated carcinoma. Several weeks after craniotomy, the patient developed respiratory failure and died. While it appears that the esophageal cancer metastasized to the brain, there is the possibility of other undetected primary tumor with metastasis to the brain. Nonetheless, the endoscopic appearance and clinical presentation of this case are unusual and noteworthy. PMID- 10826017 TI - Tuberculosis at the millennium (yes, we should worry). PMID- 10826018 TI - Highlights of the 13th annual scientific meeting of the Association for Academic Minority Physicians, 1999 PMID- 10826019 TI - Gastroesophageal reflux disease: pathophysiology and pharmacology overview. AB - Gastroesophageal reflux disease (GERD) is one of the most frequently encountered illnesses in the Western Hemisphere. GERD encompasses a spectrum of disorders in which reflux of gastric content into the esophagus causes symptoms and/or damage to the esophagus, oropharynx, or respiratory tract. This article provides a brief update on the pathophysiology and pharmacology of drugs used for the treatment of GERD. The etiology of GERD is multi-factorial and is believed to be principally a consequence of altered motility states in the esophagus and stomach. The drugs used for the treatment of GERD are continuously evolving, but as yet no drug has been shown to cure this chronic, relapsing disease. Antacids, prokinetics, and gastric antisecretory agents are the principal drugs currently used to treat GERD in conjunction with life-style modifications. Due to their ultrashort duration of buffering action, antacids are primarily used as self-medication for temporary relief of mild GERD symptoms. The prokinetic drug cisapride effectively resolves symptoms and heals mild-to-moderate esophagitis, with efficacy similar to that of the histamine H2-receptor antagonists. H2-receptor antagonists exhibit moderate inhibition of gastric acid secretion and are effective for resolving symptoms and healing mild-to-moderate esophagitis. In addition, H2-receptor antagonists slightly augment the therapeutic efficacy of cisapride for healing mild-to moderate esophagitis. However, use of H2-receptor antagonists at higher doses and higher frequency approaches the efficacy of proton pump inhibitors in healing erosive esophagitis. Given their potent and long-lasting acid-reducing efficacy, proton pump inhibitors have become the drugs of choice for many patients with GERD. Despite progress in the medical treatment of GERD, there are still several unresolved questions relating to cost-effective strategies with specific drugs, how long pharmacologic therapy should be maintained, and when surgical intervention is warranted. Additional studies are clearly needed to address the unresolved treatment issues in GERD. PMID- 10826020 TI - Gastroesophageal reflux disease: diagnosis and management. AB - Gastroesophageal reflux disease (GERD) is defined as the presence of symptoms and/or tissue damage resulting from the reflux of gastric contents into the esophagus. It occurs as a result of transient or persistent reduction in lower esophageal sphincter (LES) pressure, influenced to some degree by the presence or absence of a hiatal hernia, and the failure of the usual clearance mechanisms that normally rid the distal esophagus of noxious materials. Heartburn and regurgitation are the most common symptoms, but extraesophageal symptoms related to aspiration may occur. The clinical presentation itself is often diagnostic, but techniques such as endoscopy, barium swallow, and pH monitoring are confirmatory. Management generally involves life-style changes with or without added pharmacologic therapy. A small percentage of patients require antireflux surgery. Pharmacologic management options include acid-neutralizing agents such as antacids and alginate, prokinetic agents such as metoclopramide and cisapride, and antisecretory drugs such as the histamine H2 blockers and the proton pump inhibitors. Once the patient is healed pharmacologically, maintenance pharmacologic therapy is necessary to prevent relapse. Antireflux surgery may be indicated in patients whose diagnosis is clear, who respond well to pharmacologic therapy, but who, for one reason or another, are not candidates for long-term pharmacologic management. Preventing the advent of Barrett's esophagus is one goal of therapy, because of the risk of developing adenocarcinoma of the esophagus. The management of Barrett's esophagus is discussed. PMID- 10826021 TI - Gastroesophageal reflux disease: surgical considerations. AB - Technical advances are changing the attitude toward surgery as a last-resort treatment option for gastroesophageal reflux disease (GERD). Although a number of effective medications are currently available to manage GERD, surgery is definitive therapy, and its results are long-term, eliminating both medication compliance problems and the high cost of lifelong drug therapy. Numerous procedures are available for GERD; laparoscopic Nissen fundoplication is emerging as highly suitable and successful for most patients with simple GERD. For the best clinical result, however, the procedure must be tailored to the patient's disease and to the preoperative study results of esophageal pathology and function. The multidisciplinary collaboration of the gastroenterologist, radiologist, and surgeon will be key to the future management of GERD. PMID- 10826022 TI - Managed care, MSOs and unions. What is really going on here? PMID- 10826023 TI - Shifting paradigms in periodontal disease. Implications for prevention in the elderly. AB - Our understanding of periodontal diseases has evolved considerably. No longer are the notions of universal susceptibility to plaque and calculus with automatic reliance on mechanical debridement valid. The interplay between specific bacteria and the immune response within a complex context of risk factors forms the new paradigm of periodontal disease pathogenesis. New approaches to treatment and prevention and their implications in the elderly will be discussed. PMID- 10826024 TI - Considering a new paradigm in caries diagnosis. Towards a more conservative approach to treatment. AB - Advances in the understanding of caries and the introduction of new treatment techniques have led to the development of a new approach, or new paradigm. This paper contrasts the new paradigm with its predecessor, discusses the implications of both for the diagnosis and treatment of caries, and describes impediments to the adoption of the new paradigm. PMID- 10826025 TI - Evidence-based dentistry. It's development and use in clinical practice. AB - Science and technology are developing so rapidly that dental practitioners have to constantly assess whether the materials and procedures they are using in their practices still represent the best treatment for their patients. But busy practitioners have little time to research these questions on their own, and their attempts to do so are frequently handicapped by the lack of in-depth training in the skills required to distinguish good science from poor science. The emergence of evidence-based dentistry, a process for ensuring that the treatment we provide our patients is justified by the best available scientific evidence, holds promise for making this problem more manageable. PMID- 10826026 TI - Why not Medicaid dentistry? AB - Charitable dental services and Medicaid dentistry are two components of services provided to patients with limited financial resources. An approach to making the case for improvements in Medicaid dentistry may best be made by increasing the public's awareness of the charitable services provided by individual dentists. PMID- 10826027 TI - The professional is political: an interpretation of the problem of the past in solution-focused therapy. AB - This paper proposes that therapists subject their work to radical doubt designed to uncover and interpret the micro-politics that in here in professional knowledge and practice. Using Orlie's notion of trespass, the approach to the past employed by solution-focused therapies is examined. These trespasses involve the therapy's silence about the meaning of injustice; its role in relation to social movements; and its potential to reproduce relations of domination. A new conception of ethics is proposed that takes into account the inevitable presence of the political as it fashions the history that conditions us as human beings. PMID- 10826028 TI - "Computer addiction": a critical consideration. AB - Problems related to intemperate Internet use are examined within the conceptual framework of addiction. It is argued that empirical support for the construct validity of computer addiction has yet to emerge, that defining the construct as a unique psychiatric disorder is therefore premature, and that, in most cases, excessive computer use may be symptomatic of other, more primary disorders. Greater caution and rigor are urged in investigating and treating problems related to intemperate computer use. PMID- 10826029 TI - Psychosocial intervention in stroke: Families in Recovery from Stroke Trial (FIRST). AB - A family-focused psychosocial intervention for stroke survivors is described and illustrated with case studies. It is designed to improve functional recovery through four specific pathways: increased knowledge, efficacy, and control through stroke education; optimized social support; increased network cohesion; and improved problem-solving abilities. Rationales for these pathways are presented and methods of implementing them discussed. PMID- 10826030 TI - Cultural determinants in the treatment of Arab Americans: a primer for mainstream therapists. AB - As the population of Arab Americans grows, so does their presence among mental health clientele, creating a need among clinicians for information about these clients. The broad lines of Arab culture are delineated: its roots, language, religion, and political history; patterns of immigration to the United States; and the salient differences between Arab culture and the dominant U.S. culture. The effects of negative stereotyping and discrimination against Arab Americans are examined, as are specific clinical issues in treating them. Recommendations for more culturally sensitive treatment are enumerated. PMID- 10826031 TI - Attachment experiences transformed into language. AB - This exploratory study examined the linguistic components of adults' narratives on their attachment experiences, focusing on the process of transformation of the nonverbal experience into language. Securely attached mothers were found to differ significantly from insecurely attached mothers on cognitive ability to connect between experience and language. Results support the notion that language becomes the dynamic tool through which the individual negotiates meaning and transforms the nonverbal organized pattern of relatedness. Applications for therapy and directions for further research are discussed. PMID- 10826032 TI - Employment outcomes in family-aided assertive community treatment. AB - Family-aided assertive community treatment (FACT) was enhanced by adding vocational specialists to help persons with severe mental illness obtain competitive employment. Results were then tested against those of conventional vocational rehabilitation (CVR). The FACT cohort demonstrated significantly better employment rates than did the CVR, while negative symptoms declined in the former and increased in the latter. No evidence was found that competitive work presented a significant risk for relapse. PMID- 10826033 TI - Social/emotional intelligence and midlife resilience in schoolboys with low tested intelligence. AB - Seventy-three inner-city boys with a mean IQ of 80 were followed prospectively from age 14 until age 65. Their adult adjustment was compared to a socioeconomically matched sample of 38 boys with a mean IQ of 115. Although childhood social disadvantage did not distinguish the groups with low and high IQs, half of the low-IQ men enjoyed incomes as high and had children as well educated as did the high IQ men. These resilient low-IQ men were more likely to be generative, to use mature defenses, and to enjoy warm object relations than the high IQ group as a whole. PMID- 10826034 TI - Effects of early intervention on psychiatric symptoms of young adults in low-risk and high-risk families. AB - Effects of early risk and intervention on the psychiatric symptoms of young adults were studied. By means of a weighted risk index, 160 families with a baby born in 1975-1976 were classified as being at low or high risk. Half of the families attended a five-year family counseling program, the other half served as a control group. At age 20-21 years, the young adults from the counseled families had significantly fewer psychiatric symptoms than did those from the control families. A significant interaction between family risk and counseling was found. Implications for preventive intervention efforts are discussed. PMID- 10826035 TI - Preference for violent electronic games, self-concept, and gender differences in young children. AB - Electronic game-playing has been linked to adjustment problems in player subgroups. This study examined relationships among time commitment, gender, preference for violent games, and self-concept in 364 fourth and fifth graders. Main effects were identified for game preference and gender, with stronger preference for violent games being associated with lower self-perceived behavioral conduct. Implications for future research are discussed. PMID- 10826036 TI - Coping with psychotic symptoms in the early phases of schizophrenia. AB - How people diagnosed with schizophrenia cope with positive symptoms after their first hospitalization is explored, along with the relationship of their coping strategies to their psychosocial functioning. The strategies most frequently endorsed were cognitive in type, while those considered most helpful were behavioral. Respondents identifying an active strategy as most helpful displayed better psychosocial functioning at 24-month follow-up. PMID- 10826037 TI - Comorbidity of substance use and post-traumatic stress disorders in a community sample of adolescents. AB - A study of 384 18-year-old adolescents living in the community demonstrated a frequent co-occurrence of substance use disorders and post-traumatic stress disorder. Multiple pathways appeared to lead to this comorbidity, which was associated with widespread psychological impairment that might have serious developmental consequences. Implications for research and practice are discussed. PMID- 10826038 TI - Exposure to community violence and post-traumatic stress symptoms: mediating factors. AB - Children's perceptions of safety and family conflict were examined as mediators of the relation between exposure to community violence and post-traumatic stress symptoms. Results indicated that exposure to community violence was related to perceptions of decreased neighborhood safety and increased family conflict, and that this, in turn, mediated the impact of violence exposure on children's post traumatic stress symptoms. Implications for research are discussed. PMID- 10826039 TI - Trauma-related symptomatology among children of parents victimized by urban community violence. AB - The effects of parents' victimization by community violence on children's psychological functioning was examined in a group of 30 6-12-year-old African American children. In comparison with 30 demographically similar children in the same urban community whose parents had never been victimized, the victims' children exhibited higher levels of distress symptoms. Implications for research and practice are discussed. PMID- 10826040 TI - Prenatally drug-exposed toddlers: cognitive and social development. AB - In a replication of an earlier study, 122 prenatally drug-exposed toddlers were assessed on four cognitive scales and rated by their caregivers on socialization, communication, and daily living skills. A significant number of the sample performed below age expectancy on the cognitive scales and on at least two of the three domains rated by caregivers. PMID- 10826041 TI - Dentistry for babies: a preventive protocol. AB - The objectives of this program were to educate the parents in the importance of preventive care and to establish protocols for oral health treatment and maintenance that reflected the oral disease risk of the infant. Since its inception, more than 1,000 infants have been enrolled in the program and compliance has been excellent. PMID- 10826042 TI - Cost-effectiveness study of a school-based sealant program. AB - A nonprofit private dental clinic provides free dental treatment for children up to the age of eighteen years. In order to expand its services to reach more children, a school-based sealant program using dental auxiliary personnel was organized. This paper evaluated the performance of the school-based program during 1991 by comparing the costs of the school-based program with the costs of the sealants placed in the clinic in twelve months. Costs were calculated based on 1991 expenditures, and effectiveness rates were estimated from dental literature on sealants. The cost of saving one tooth-surface from decaying within a six-year period at the school and the clinic was $65 and $42 with an average sealing time per tooth surface of 18 and 12.5 minutes, respectively. Despite the relatively lower cost of personnel, the school program cost was 35 percent higher than at the clinic. Nevertheless, if hidden costs, such as transportation, time off work, and waiting time for those attending the clinic were considered, costs could be comparable or even higher for the clinic. The main goal of this paper is to explore a methodology to compare programs of different nature, and critically evaluate the results. PMID- 10826043 TI - An increasing number of your pediatric patients may have asthma: the demographics of asthma. AB - In light of the growing prevalence of children with asthma, dental practitioners should increase their awareness of the demographic characteristics of the disease and the associated use of health services. A review is provided of 1) the distribution of the disease in the pediatric and general populations, 2) the number of physician visits, 3) emergency room and hospital usage, and 4) mortality patterns. PMID- 10826044 TI - Childhood asthma and dental erosion. AB - The aim of this investigation was first to assess the prevalence of asthma in a random sample of fourteen-year-old children in Birmingham UK. Secondly to assess the levels of dental erosion in these children, and thirdly to see whether there was any correlation between children with asthma and the levels of dental erosion. A random sample of 418 children from twelve secondary schools was examined, 209 were male and 209 female. The level of tooth wear was recorded using a modification of the Tooth Wear Index of Smith and Knight. The prevalence of asthma in this group was 15.8 percent (66 children out of 418); the levels of dental erosion in children with asthma were higher. PMID- 10826045 TI - Pediatric dental practice: reconstruction or disintermediation. AB - The objective of this study was to evaluate ongoing changes in contemporary pediatric dental practice. The impact of demographics, gender, generational values and debt load of the 1990s practitioner is reviewed. The role of overhead expenses, third party payers, business practices, dental management service organizations and business training of practitioners are analyzed. The influence of technological changes on clinical practice is described. In conclusion, there is a need for zero-basing and reconstructing pediatric dental practices. PMID- 10826046 TI - Bond strengths of glass ionomer restoratives to primary vs permanent dentin. AB - An in vitro experiment was conducted to evaluate the shear bond strength of a conventional GIC (glass ionomer cement) and a RMGIC (resin modified glass ionomer cement) when applied to dentin of primary and permanent teeth. Results show that the bond strength of the RMGIC was more than thrice that of the conventional GIC. Fracture analysis showed that the bond failures were cohesive in the cement. PMID- 10826047 TI - In vitro study of antibacterial and antiadhesive activities of fluoride containing light-cured fissure sealants and a glass ionomer liner/base against oral bacteria. AB - The aim of this study was to estimate the antibacterial and anti-adhesive properties of light-cured pit-and-fissure sealants containing fluoride (Teethmate F-1, TF; UltraSea-1 XT, US) or not containing fluoride (Teethmate A, TA)- and glass ionomer liner/base containing fluoride (Vitrebond, VB) against oral bacteria in vivo. VB inhibited the growth of S. mutans, S. oralis and L. casei aerobically and anaerobically, and TA and US inhibited the growth of only S. mutans aerobically. The numbers of adherent cells mediated by insoluble glucan for all materials were lower than that for the control tooth. The numbers of adhesional cells not mediated by insoluble glucan were larger than that for the control tooth. VB showed the highest concentration of fluoride released throughout the four-week observation period, and had the highest pH value. In this study, VB showed superior antibacterial action and fluoride-release compared with the other three materials. PMID- 10826048 TI - Pain related to different ways of minimal intervention in the treatment of small caries lesions. AB - Pain during invasive treatment of dental caries is a common phenomenon, if no local analgesia has been used before cavity preparation. Atraumatic restorative technique (ART) is a suggested procedure which is at least less traumatic for the patient. Although the ART approach has been received well by both children and adults who belong to population groups hardly ever exposed to regular oral health care, it has not yet been proven that this particular procedure really causes less pain, compared to more conventional techniques with rotating instruments. In the present study, pain was reported in connection with tooth restoration in 19.3 percent of the cases, when the ART technique was used, which is significantly less than with a conventional restorative technique (35.7 percent). Finally the results show a clear relationship in the pain reports between the first and the following treatments in both ART and the conventional technique groups. PMID- 10826049 TI - A survey of oral biopsies in Brazilian pediatric patients. AB - This paper presents a review of 1018 oral biopsies in pediatric patients from the Oral Pathology Service, Minas Gerais Federal University, Brazil. The lesions were divided into ten main categories. The most common oral lesions in this study were follicular cyst in the maxillary anterior region, followed by inflammatory fibrous hyperplasia in the same region, and mucocele in the lower lip. Cysts of the jaws and oral soft tissues comprised 26.1 percent of total oral biopsies. The importance of these findings in oral diagnosis is discussed. PMID- 10826050 TI - Dental caries and fluorosis prevalence study in a nonfluoridated Brazilian community: trend analysis and toothpaste association. AB - The main purpose of this study is to compare data from previous surveys to current prevalence rates of dental caries and dental fluorosis in eleven- and twelve-year-olds in a non-fluoridated Brazilian community (< 0.2 ppm F). This study also assesses the possible association between use of fluoride toothpaste and the prevalence of dental caries and dental fluorosis. The sample subjects are randomly selected schoolchildren who were examined with a dental probe and buccal mirror under natural light. The intra-examiner error was calculated, using Kappa statistics (K tau 0.85). The results showed that between 1991 and 1997 there was a 56.7 percent decrease in the prevalence of dental caries and an 80.1 percent increase in dental fluorosis. Children with dental fluorosis were 1.75 times more likely to be free of caries (OR = 1.75-CI:0.43, 6.68). Children who started using fluoride toothpaste before the age of three were 4.43 times more likely to have dental fluorosis than those who started using it after the age of three (OR = 4.43-CI:0.51, 99.61). The results of the cross-sectional surveys conducted with schoolchildren in 1991, 1995, and 1997 suggest a continuing decrease in the prevalence of dental caries and an increase in the prevalence of dental fluorosis in this nonfluoridated Brazilian town. PMID- 10826051 TI - Radiographic evaluation of third-molar development in a group of Turkish children. AB - The aim of this investigation was to evaluate the crypt formation and calcification of the maxillary and mandibular third molars according to age and gender on panoramic radiographs in a group of Turkish children. Four hundred panoramic radiographs of 188 female and 212 male dental patients, ages six to thirteen years, were examined by two observers. The interrater reliability was tested by Kappa statistics. The earliest age for maxillary third molar crypt formation was eight years; mandibular third molars could be seen radiographically as early as seven years. PMID- 10826052 TI - Evaluation of calcium hydroxide and zinc oxide eugenol as root canal filling materials in primary teeth. PMID- 10826053 TI - Coffin-Lowry syndrome and premature tooth loss: a case report. AB - Coffin-Lowry syndrome was first described by Coffin in 1966 and later by Lowry in 1971. Several oral manifestations have been described previously. We report a case with additional information on the histology of prematurely exfoliated teeth. PMID- 10826054 TI - [The cell as a gravity-dependent biomechanic system]. AB - In the period of 1995-1997 experimental and theoretical studies with various biomechanic objects, i.e. individual cells and cell associations, were performed under changed gravity (0.00001-5 g). Experimental investigations were conducted using clinostats and centrifuges to model effects of hypo- and hypergravity, and aboard space vehicles in real microgravity. Cell cultures in vitro including fibroblasts and osteoblasts on a solid glass or plastic substrate served as objects of the studies. Changes in value and direction of the gravity vector were found to modify the morphophysiological characteristics of cells: structural organization (spatial rearrangement of the intracell component, changes in forms, sizes and quantity of cells) and functional activity (alterations in energy expenditure and intensity of intracellular metabolism). The data suggest that there should be mechanisms of gravitational sensitivity in living systems on the cellular level. As was stated, sensitivity of unicellular free-living organisms to gravity is mostly defined by the motor activity determined by the level of general metabolism. Morphological characteristics (form, size and mass) are of secondary importance. Theoretical analysis resulted in correction of one of the principle postulates of gravitational biology stating a direct link between size (mass) and gravitational sensitivity of organism. Described were consistent patterns of growth, development, and behavior of unicellular cultures in gravitational fields. Strengthening of the force of gravity (hypergravity) leads to eventual deceleration of cell growth and diminution of biomass gain. On the other hand, the spaceflight environment (microgravity) stimulates growth mechanisms. In our opinion, behind these gravitational effects are altered levels of energy spent by cells to overcome the force of gravity. Opposite trends were observed in experiments with cell cultures in vitro. During space microgravity, fibroblast cultures on the solid substrate decreased the growth rate, and inhibited cell division and migration within the substrate. Compared to the Earth's gravity, under elevated gravity these parameters were noticeably higher. It was demonstrated that the main cause of the unfavorable effects of space microgravity on the cellular level is decay in the adherence of cells to the substrate. Explored were also the most probable mechanisms of the effects of changed gravity on the cell as a biomechanic structure. Specialized and non specialized graviceptors of various types of cells were crypt-analyzed and classified. In future, investigations should be angled for elucidation of the role of intracellular components in perception and implementation of the gravitational stimulus, and description of quantitative characteristics of energy exchange and metabolism in cells as a function of gravity force and direction. PMID- 10826055 TI - [Ethics of space life science research with animals]. AB - The authors present a historical sketch on the ethics of use of animals in life sciences experiments. Experiments with animals are necessary to expand knowledge in the field of life sciences including the space ones, and health service. It is emphasized that these experiments must be performed with observance of certain moral rules and utilize minimum animals suffice to obtain statistically significant data. The first draft of SRC-IBMP Guidelines on the use of laboratory animals has been published. PMID- 10826056 TI - [Problems of environmental compatibility of the Russian and US segment of the International space station]. AB - The problem of environmental compatibility of the ISS Russian and U.S. segments arises from chemical and biological inequality of their atmospheres and is attested by the data of observations during operation of Russian orbital stations Salyut and Mir. Efforts must be made to develop unified limits, and common procedures and requirements for evaluation of ISS materials and equipment by experts of the international partners. PMID- 10826057 TI - [Aspects of developing a unified information system of space crew medical support based on networking technologies]. AB - Computerization of national and international space research projects, real-time data management, improvement of the quality of medical support to space crew through more extensive use of video, audio and telemetric information put in the forefront the necessity to integrate individual experts and agencies in a single network. Described is the structure of the operative network mounted at the Russian Federation State Research Center--Institute of Biomedical Problems. The authors discuss issues related to networking participants in medical support of manned missions, and a biomedical data exchange format. Developed was a prototype of an on-line Web-server. Prospect for advancement of the informational network is outlined. PMID- 10826058 TI - [Information technology of diagnostics of the operator's functional status]. AB - Consideration has been given to the theoretical and applied aspects of regulation of the operator's functional state and its impact on work quality from the standpoint of preparation for and implementation of space mission. Determinations of the reflectory and regulatory functions of the operator's state are given and the neurophysiological mechanism of the latter is characterized. Analyzed were fresh methods of controlling the states of healthy persons and patients subjected to psychogenic depression via remote monitoring of voice, motor activity, etc., and recording of electrical indices (EEG, autonomous functions, etc.). Peculiarities of modifications of errors made by human subjects in identification of masked visual images within the "fatigue--negative emotions" range were investigated. PMID- 10826059 TI - [The mechanisms of insulin resistance during hyperthermia]. AB - Consideration is given to the possibility and mechanisms of insulin resistance as a function of human tolerance to hyperthermia. Poor tolerance of hyperthermia was observed in persons with initially altered metabolism. Administration of sugar test by these persons revealed high levels of insulin incretion and low insulin reactivity of tissues or insulin resistance. Negative effects of extended period of hyperthermia that may bring about pathologic pancreatic changes even in essentially healthy persons are reviewed. Following LCE and rehabilitative therapy, 60% of aviators were medically certified without change in fitness category. PMID- 10826060 TI - [The influence of extended exposure in artificial hypercapnic environments on the function of external respiration and gas exchange]. AB - Effects of artificial hypercapnic atmosphere were studied in a series of ten 40-d experiments with participation of 42 volunteers aged from 20 to 61 years. Partial pressure of carbon dioxide was maintained at 27 +/- 2 mm Hg (3.4-3.8% CO2); partial pressure of oxygen was in the range of 122-136 mm Hg. Barometric pressure, air temperature and humidity had normal values. Results of the experiments were subjected to the variation analysis. Continuous breathing the gas mixture with specified PCO2 increased CO2 sensitivity of the breathing center in all age-groups both at rest and during exercise. The ventilatory reaction of subjects aged 41-61 grew to a less degree as compared to the subjects below 40. Medical selection of members for space crew to go to a distant voyage should take into consideration the parameters of oxygen demand and the volume of pulmonary ventilation per a time unit. PMID- 10826061 TI - [The build-up of human reserve potential by exposure to intermittent normobaric hypoxia]. AB - Effects of intermittent hypoxia at normal pressure (NPH) on physical and mental performance were studied in 32 essentially healthy human subjects. The 24-d NPH cycle was found to increase the maximal aerobic capacity as well as physical and mental performance. Growth of reserve and efficiency of the main physiological systems stipulated this gain in physical and mental performance. The authors make a presumption that NPH can be a part of preflight training, a measure to maintain an adequate level of human performance during orbital mission, and a method of rehabilitation post flight. PMID- 10826062 TI - [Response of siphoneal alga (Vaucheria sessilis) to the gravity factor]. AB - Growth and development of siphonal alga Vaucheria sessilis under changed gravity and the role of cytoskeletal structures in gravitational response were studied. Hypergravity (3 g) and "hypogravity" were generated by slow clinostating at 7-8 rev/min and rapid clinostating at 35, 70, 135 rev/min, respectively. The experiments in microgravity were flown aboard biosatellite Bion-11. As was shown, V. sessilis responded to changed gravity by inhibition of rate of growth and increasing the number of nuclei in the strand as a result of activation of mitotic processes. Modulation of the course of the gravitational response with anticytoskeletal agents points to involvement of the cytoskeleton. The cortical circuit of actin microfilaments and the cytoplasm stream directed by this structure appear to be the most sensitive to changes in gravity. Gravity sensitive V. sessilis is another promising object of research in the field of gravitational biology. PMID- 10826063 TI - [Experimental study of the root supply system with periodic water return designed for space greenhouses]. AB - To improve reliability of plant's moistening and aeration control in microgravity, an original root supply system with a periodic return water flow has been designed and tested in laboratory. For 30 days crops of Pekinese cabbage (Brassica pekinesis (Lour Rupr), Khibini sort) were raised in the test bench which allowed adjustment of water potential in the root zone within a preset range. A three-step water potential control algorithm included water injection with a pump-dispenser, a pause, and water sucking back to a desired value of water potential. The following parameters of the control cycle were selected in a series of two experiments: time of water injection (2.5 hr) and return (1.5 hr), and a pause of 8 and 20 hr, respectively. Magnitude of water potential about the root module axis was controlled in the range from -1.3 kPa to -3.0 kPa in both experiments and maintained at -1.3 kPa in the control. The root modules consisted of porous metaloceramic tubes wrapped in fibrous ion exchanging cloth and a light proof film with planting slots on top. In the first experiment, plant characteristics were comparable to the control. The developed procedure and technology can be used to provide favourable moisture-air conditions in the root zone. By and large, the system of root nutrition with a periodic water return has demonstrated high capacity during the ground-based cultivation of plants. To use this system in space greenhouse, it is necessary to specify operational parameters for the microgravity environment. PMID- 10826064 TI - [The balance of harmful trace contaminants between the air humidity condensate and air in a simulator of the Mir orbit station moisture condensation unit]. AB - Subject of the investigation was the balance of harmful trace contaminants (HTC) between the air moisture condensate and air in a simulator of the MIR moisture condensation unit. Experiments involved various classes of water-solvent compounds including alcohols (C1-C4), ketons (C1-C2), aldehydes (C1-C2), fatty acids (C2-C4), esters (acetates C4-C6), and ammonium. For most of the compounds, removal efficiency correlates with air humidity and virtually does not depend on the HTC concentration within the range of 0.25 to 59.1 mg/m3. PMID- 10826065 TI - [Laparoscopic cholecystectomy in the treatment, rehabilitation and clinical health certification of the flying personnel with cholelithiasis]. AB - Applicability of laparoscopic cholecystectomy (LSCE), a modern approach to the treatment of cholelithiasis, to rehabilitation, and military medical and military aviation certification was evaluated. Since 1994, 303 LSCE operations, including to 97 patients-aviators, have been performed by way of penetrating the abdominal wall with a standard laporoscopic stand (K-Chtorz, Germany). After introduction of LSCE, only 9 operations (2.9%) were fulfilled with the conventional technique. Immediate results of treatment of aviators were satisfactory and characterized by a good cosmetic effect, more rapid recovery of the motor activity, minimal complications (2.2%), absence of severe post-operational pancreatic/biliary dysfunctions, and expedited course of treatment. PMID- 10826066 TI - [Electrical stimulation of the visual analyzer in the rehabilitation of flying personnel with abnormal refraction]. AB - The authors propose an original technique of rehabilitative treatment of visual disorders in flying personnel, i.e. transconjunctival electroophthalmostimulation, which utilizes a specially developed ophthalmoscopic electrostimulator (ESOF-1). The technique was applied to 312 flyers with deteriorated visual acuity due to refraction abnormalities. After the course of 10 stimulation sessions, the far- and near-sight acuity was noted to improve by 48% and 38%, respectively, accommodation volume by 51%, accommodation reserve by 65%, and contrast sensitivity by 23%. This gain in vision remained after one year since treatment but slightly declined (23-38%) by the end of the second year of follow-up. Moreover, the results of therapy were not dependent on severity and type of refraction, or age of a patient. Without treatment, 56% of pilots and 68% of navigators would have been grounded for the reason of bad vision. Transconjunctival electroophthalmostimulation enabled 90% of the flyers to continue their career. PMID- 10826067 TI - [A model system for the assessment of the stability of eukaryotic genes and expressed proteins in extended space flight]. AB - Evidence was obtained that the changed gravity, tension profiles of the magnetic fields inside the orbital station and other spaceflight factors (SFF) substantially influence the cell genome and synthesis of recombinant proteins. The authors proposed a technique of fixation of possible alterations in genes and expressed recombinant proteins in strains-producers of human alpha-interferon: HuIFN-alpha 2b, HuIFN-alpha 8a, HuIFN-alpha 10a, and HuIFN-alpha 14a. Spaceflight factors were simulated by way of gamma-irradiation at 10 Gy and 50 Gy by a cobalt unit, and centrifugation of samples at 10 g and 50 g. Cultivation of the strains producers during the SFF simulation yielded HuIFN-alpha proteins with altered functional characteristics. Following exposure to simulated SFF, strains producers expressed HuIFN-alpha that preserved a high titre of antiviral activity (AVA) but suppressed the antiproliferative activity (APA) inferring some structural/functional shifts in the HuIFN-alpha molecule due to, presumably, mutations in the active center responsible for APA. Determination of species specificity of the HuIFN-alpha recombinant proteins following exposure to SFF revealed dissociation of cross-AVA in homologous and heterologous cell cultures which can be also attributed to the structural/functional shifts in the HuIFN alpha molecule. These changes can be localized in the receptor cluster of molecule or consequent to modification of the center defining HuIFN-alpha species specificity. The proposed simulation system allows fixation of shifts in the HuIFN-alpha structural/functional characteristics and investigations of the stability of eucaryotic genes in long-term space flights. PMID- 10826068 TI - [The use of antimicrobial stabilizers in biochemical research]. AB - The ability of highly active antimicrobial biochemical stabilizers to sustain samples (T = 4 degrees C to 8 degrees C) till biochemical analysis in 20-50 days was tested. A new generation of delayed biochemical assays with the use of these stabilizers can be invoked in metabolic studies of cosmonauts and aviators, personnel of atomic submarines, pole explorers and other occupational groups. PMID- 10826069 TI - [Bioethical issues of medical research in the interests of military and aerospace medicine]. PMID- 10826070 TI - [At the cradle of space medicine]. PMID- 10826071 TI - [From the protection of the crew on board the place with nuclear power installation to the radiation protection on board the space vehicle]. PMID- 10826072 TI - New developments in type 1 (insulin-dependent) diabetes. AB - The Diabetes Control and Complications Trial has conclusively demonstrated that improved metabolic control leads to reduction in the rate of microvascular complications of diabetes. In order to allow patients to achieve improved metabolic control, much research has focused on improved methods of glucose monitoring and more physiologic ways of insulin delivery. The 2 most promising methods of minimally invasive blood glucose monitoring are the Glucowatch, using the technique of reverse iontophoresis to measure interstitial fluid glucose levels every twenty minutes and an implantable sensor, in which a catheter resembling that used for insulin delivery through a pump is impregnated with glucose oxidase at the tip. This device monitors blood sugars every few minutes, but like a holter monitor, must be downloaded in the physician's office. Still under development are (1) implantable subcutaneous sensors with a high and low blood glucose alarm and (2) sensors in which the patient will be able to download the data using a home PC. Advances in insulin delivery have included the availability of new insulin analogs which more closely simulate endogenous insulin release, with rapid acting analogs simulating the increase in insulin production that normally occurs after meals. Phase III clinical trials are in progress of a long-acting basal insulin without peak actions to simulate the low dose continuous production of the insulin which normally inhibits hepatic glucose production. In addition, use of the insulin pump has increased markedly since publication of the DCCT with the greatest increase being among adolescents. In addition to advances in treatment of diabetes, research has continued on curing the disease using islet cell transplantation and preventing the disease with agents such as insulin (DPT-1 Trial) and nicotinamide (ENDIT). This article provides an overview of recent advances in diabetes management and prevention. PMID- 10826073 TI - Recurrent abdominal pain in children--a retrospective study of outcome in a group referred to a pediatric gastroenterology practice. AB - Recurrent abdominal pain (RAP) affects a significant number of children each year. We reviewed our experience over a 2-year period to determine the outcome of patients who were referred for pediatric gastroenterology consultation. We identified 356 patients, 149 (42%) male and 207 (58%) female. All patients underwent a thorough interview and complete physical examination. Patients suspected of having irritable bowel syndrome (IBS) were treated as such without further initial evaluation. Others underwent an initial blood and urine evaluation. When these initial screening studies were negative, additional studies were performed including abdominal ultrasonography, radiography, and/or endoscopy of the upper gastrointestinal (GI) tract if the history suggested a possible diagnosis that could be excluded or confirmed by such tests. There was no identifiable diagnosis in 43.5% of the patients studied. IBS was diagnosed in 25.8% of all patients. Constipation was diagnosed in 3.7%. Miscellaneous causes, including GI mucosal lesions, and renal and pancreatic disorders were found in an additional 27% of patients. In a follow-up survey, more than 70% of the treated respondents were improved (i.e., their RAP had resolved or was markedly improved). We conclude that most children with RAP have a functional disorder. Patients with an organic cause for pain can be identified and treated in a cost effective manner with carefully planned evaluation. PMID- 10826074 TI - Relationship between scholastic and health behaviors and reading level in adolescent females. AB - This was a study of 102 adolescent females, 12-20 years of age, presenting to a central city clinic for medical care. Participants completed an oral questionnaire that included demographics, and questions regarding scholastic history, sexual behavior, and substance use. Each subject completed the Accuracy Level Test (ALT), a reading test. The subject's reading test grade level was subtracted from her appropriate grade in school to give a reading delay level (RDL). The mean reading grade level for all subjects was 6.7 +/- 2.6 and the average reading delay was 4.5 +/- 2.5 grades. Poor school attenders had greater reading delays (5.8 +/- 3.4 grades behind vs. 4.3 +/- 2.2 for good attenders p < 0.04), and those who repeated grades were also significantly delayed (5.5 +/- 2.4 grades behind vs. 3.7 +/- 2.3, p < 0.0007). Previously pregnant students had a greater delay in reading level than their nonpregnant peers (5.2 +/- 2.3 vs. 3.9 +/- 2.5 respectively p < 0.01). Delayed reading levels appear to correlate with some risk behaviors. The ALT may serve as a tool to identify high-risk patients who need more intensive clinical intervention. PMID- 10826075 TI - Parental attitudes toward BB and pellet guns. AB - As well as describing our pediatric BB and pellet gun injuries and the circumstances surrounding these injuries, we also evaluated parental perceptions of the dangers of BB and pellet guns. A convenience sample of three groups of parents and their children presenting to a Midwest, urban, children's hospital emergency department was prospectively enrolled. The three groups of parents included the injured group, which consisted of the parents whose children had been injured by BB or pellet guns; the gun group, which consisted of the parents who allowed their children to possess BB or pellet guns but had not sustained injury from these guns; and the no gun group, which consisted of the parents who did not allow their children to have these guns. All parents completed a survey concerning their attitudes toward BB and pellet guns. Twenty-eight parents completed questionnaires in each of the three groups. Most BB and pellet gun injuries occurred in adolescent males at home without adult supervision and were inflicted by a friend or by themselves. The injured group and the no gun group viewed BB and pellet guns as significantly more dangerous than the gun group. Parents who allow their children to have BB or pellet guns appear to misperceive their potential for injury by allowing their children to use these guns in an unsafe manner. Clinicians must educate parents about the significant potential for injury of nonpowdered guns. PMID- 10826076 TI - Infant sleep position practices 2 years into the "back to sleep" campaign. AB - Since the 1992 American Academy of Pediatrics (AAP) recommendation to put babies to sleep in the nonprone position and the subsequent 1994 "Back to Sleep" campaign, the U.S. rate of sudden infant death syndrome (SIDS) has decreased more than 40%. This study reports sleep position practices in the greater Philadelphia area during 1996 and 1997. Four hundred and ten parents of infants 6 months of age or less answered a questionnaire by interview in Philadelphia clinics and private pediatric offices from December 1995 through February 1997. Sleep position practices and other SIDS risk factors were measured among demographic groups and compared with reported rates in a similar population from 1993 and 1994. Data were analyzed by Chi square after analysis of correlation coefficients. Significance is reported at p < 0.05. Seventy-two percent of all infants surveyed slept nonprone (NP) compared to 31.8% in 1993 and 59.1% in 1994. The population was 61% African-American (AA), 62% clinic patients. The breast feeding rate was 31%, maternal smoking 17%, and cosleeping 46%. AA infants (67% vs. 82%), infants receiving care at a clinic (66% vs. 84%), and infants > 3 months old (65% vs. 76%) are less likely to be placed nonprone. Most parents who place infants on their back report it was recommended by a medical professional (56%). The majority of those placing infants prone do so because their infant is more comfortable or sleeps better (65%), although 73% said their physician/nurse discussed sleep position with them. Nonprone sleeping continues to increase since the initiation of the "Back to Sleep" campaign. Disparity between some demographic groups persists. An excessive number of African-American families and clinic families still choose a prone sleep position. Many who do so cite increased infant comfort, despite knowledge of the AAP recommendation. PMID- 10826077 TI - A 9-month-old boy with fever and lethargy. PMID- 10826078 TI - Delayed presentation of pulmonary interstitial emphysema. PMID- 10826079 TI - A hand-held, reusable, flexible fiberoptic catheter to conform correct endotracheal tube placement in children: an evaluation and comparison with chest radiography. PMID- 10826080 TI - Avascular necrosis of the hip: a complication following treatment of congenital dysplasia of the hip. PMID- 10826081 TI - Dilemma faced by every admissions committee when considering the application of a reasonably acceptable minority candidate who has no personal record of deprivation. PMID- 10826082 TI - Sarcoidosis in children. PMID- 10826083 TI - What's eating you? Bedbugs. PMID- 10826084 TI - Melanoma in children. PMID- 10826085 TI - Coffee grinders assist pediatric dosing. PMID- 10826086 TI - Photo quiz. What is your diagnosis? PMID- 10826087 TI - Lichenoid drug reaction due to sildenafil. AB - Lichenoid drug eruptions are difficult to distinguish from lichen planus. Determining the offending agent is complicated by the proliferation of lifestyle medications that the patient may not consider a medication. A case of lichenoid drug eruption due to sildenafil, which was taken for sexual enhancement, is presented. PMID- 10826088 TI - Peanut allergy. AB - Peanut allergy is acute and severe with symptoms of immediate hypersensitivity. This allergy is very common, affecting 1% of preschoolers. The incidence has increased with succeeding generations, and is possibly due to the increasing exposure of children to peanuts at a young age. Diagnosis is via history, skin prick test, and serum IgE level. The mainstay of therapy is avoidance. Treatment of anaphylaxis includes epinephrine and antihistamines. Children usually will not outgrow this food allergy. Novel treatment with rush immunotherapy and enzyme potentiated desensitization is not currently acceptable. We describe a 27-month old Asian boy with a typical presentation of peanut hypersensitivity. A good understanding of the epidemiology of this illness is necessary for treatment and prevention. PMID- 10826090 TI - Onchocerciasis presenting with lower extremity, hypopigmented macules. AB - Onchocerciasis, or river blindness, is a parasitic infection caused by the filarial nematode, Onchocerca volvulus. It infects 18 million people worldwide, but is rarely seen in the United States. It is one of the leading causes of blindness in the developing world. Although onchocerciasis is also known as river blindness, it is not just a disease of the eyes, but rather a chronic multisystem disease. Clinically, onchocerciasis takes three forms: 1) eye disease; 2) subcutaneous nodules; and 3) a pruritic hypopigmented or hyperpigmented papular dermatitis. We present an 18-year-old African female with a 5-year history of asymptomatic, hypopigmented, slightly atrophic macules on her anterior tibiae. Pathology revealed a scant perivascular inflammatory infiltrate with mononuclear cells, eosinophils, and rare microfilariae in the papillary dermis. Ivermectin is the treatment of choice for onchocerciasis and was initiated in this patient. We present this interesting patient with onchocerciasis to expand our differential of hypopigmented macules, especially in the African population. In addition, we discuss both the diagnosis and the treatment of onchocerciasis in expatriate patients living in nonendemic areas. PMID- 10826089 TI - Cactus skin injuries. AB - Cactus spine injuries to the skin are usually treated by removing the visible spines and applying a topical corticosteroid. With this approach, granulomatous inflammation usually resolves in 2 to 4 months. We describe a case of a 54-year old Caucasian woman who presented for treatment of painful cactus spine granulomas on her right hand. Unroofing the granulomatous papules and removing the cactus spine fragments under a dissecting microscope, and subsequent soaking in an antibacterial solution, resulted in rapid resolution of the granulomatous lesions. PMID- 10826091 TI - Blue nevus with satellitosis mimicking malignant melanoma. AB - Blue nevus rarely develops a malignant melanoma. The loss of the regular border and the development of satellite lesions are ominous clinical indicators of the malignant change. A case is presented in which both of these clinical features- irregular border and satellite lesions--were present, but no malignant change was observed histopathologically. To our knowledge, such a single nodule with satellite lesions mimicking malignant melanoma has not been described previously. PMID- 10826092 TI - Subungual amelanotic melanoma. AB - We describe a 76-year-old white male with subungual amelanotic melanoma. The lack of pigmentation of the lesion may cause misdiagnosis and aggravate its poor prognosis. PMID- 10826093 TI - Multiple corticosteroid orally elicited allergic contact dermatitis in a patient with multiple topical corticosteroid allergic contact dermatitis. AB - Corticoid allergic contact dermatitis (ACD) may be topically or systemically elicited. Allergic contact dermatitis to topical corticosteroids is relatively common, whereas reports of orally elicited ACD to corticosteroids are rarer. Patients allergic to one corticosteroid often exhibit cross-reactivity to other corticoids. We have previously reported a 46-year-old woman with contact allergy documented by patch and provocative use testing to multiple topical corticosteroids. On further testing, she was thought to have multiple corticoid orally elicited ACD to triamcinolone, methyl prednisolone, dexamethasone, and prednisone. Oral provocation tests were performed in a single-blind fashion following the method of Alanko and Kauppinen [Diagnosis of drug eruptions: clinical evaluation and drug challenges. In, Skin Reactions to Drugs (Kauppinen K, Alanko K, Hannuksela M, Maibach HI, eds). Boca Raton, FL, CRC Press, 1998.]. The five oral corticosteroids tested were triamcinolone, methyl prednisolone, dexamethasone, prednisone, and hydrocortisone. Four of the five challenged corticosteroids (i.e., triamcinolone, methyl prednisolone, dexamethasone, and prednisone) produced a generalized maculopapular eruption in a delayed manner. The fifth challenged corticoid, hydrocortisone, had no adverse effect on this patient. This patient was unusual in that she exhibited polysensitivity to a spectrum of oral and topical corticosteroids. Hydrocortisone was identified as a corticosteroid for future clinical use. This is an important finding since corticosteroids are important emergency drugs. PMID- 10826094 TI - Malignant melanoma with zosteriform metastases. AB - Melanoma is a tumor that has a high tendency to metastasize to the skin. Zosteriform metastases are a rare form of metastasizing melanoma. We describe a 79-year-old man with melanoma presenting with a dermatomal distribution and no visceral metastases. PMID- 10826095 TI - Transverse leukonychia with systemic infection. AB - Transverse white nail bands (leukonychia) have been described in association with systemic illnesses and exposure to toxins, and medications. We describe the occurrence of transverse nail bands in two patients following acute systemic illnesses. In the first case, transverse white nail bands developed in a 30-year old human immunodeficiency virus-positive man following acute pulmonary tuberculosis. In the second case, transverse white nail bands were noted in an 80 year-old patient following Streptococcus intermedius empyema. PMID- 10826096 TI - Allergic contact dermatitis to gold. AB - Gold is a relatively common allergen that appears to induce dermatitis about the face and eyelids, as well as at sites of direct cutaneous contact. In this study, 355 patients with suspected contact dermatitis were evaluated; 17 (4.8%) were found to be allergic to gold. Fifteen of these 17 patients were re-evaluated at > 2 months after patch testing. When contact with gold jewelry was discontinued, 7 of 15 (46.7%) of the gold-allergic patients reported that their dermatitis cleared. In 3 of 7 patients (42.9%), discontinuing contact with gold jewelry was the only modification to their behavior; whereas in 4 of 7 (57.1%), discontinuing contact with gold jewelry and other documented allergens was necessary to affect resolution. Despite continuous contact with gold (jewelry and/or dental appliances), 7 of 15 (46.7%) of our patients had complete clearing of their symptoms by avoiding other documented allergens. None of our patients required the removal of dental gold. PMID- 10826097 TI - Ulcerated atrophic striae from etretinate. AB - Oral retinoids such as etretinate and acitretin are commonly associated with dose dependent, mucocutaneous side effects such as dryness, peeling, and fragility. Although these effects can be extreme in some patients and even require discontinuation of treatment, thinning of skin to the point of atrophy and ulceration has never been reported in the English literature. We present the case of a patient with psoriasis in whom ulcerated atrophic striae developed during etretinate therapy. After discontinuation of etretinate, all cutaneous ulcers resolved. Subsequently, the patient had a favorable response to oral calcitriol (1,25-dihydroxy vitamin D3), a novel therapy for psoriasis. PMID- 10826098 TI - Spontaneous apparent clinical resolution with histologic persistence of a case of extramammary Paget's disease: response to topical 5-fluorouracil. AB - An 86-year-old woman presented with a 3-year history of an erythematous axillary lesion, which was histologically confirmed to be extramammary Paget's disease (EMPD) confined to the epidermis and adnexa. Surprisingly, spontaneous clinical regression occurred in the lesion, but Paget's cells persisted within the epidermis and adnexa on histologic examination. One year of intermittent topical chemotherapy with 5-fluorouracil resulted in ulcers that were interpreted as EMPD and completely excised. Histologic examination showed a complete absence of Paget's cells. To our knowledge, only one previous report investigated apparent spontaneous clinical resolution with histologic persistence of EMPD. We emphasize that topical 5-fluorouracil cannot be considered a safe treatment modality for EMPD, but it may be useful in certain cases in which the extent of the lesions, or the general condition of the patient, advise against surgery or radiotherapy. PMID- 10826099 TI - Reproduction/developmental toxicity screening test in rats with orally administered 1-hexene. AB - This study was conducted to provide screening information concerning the potential systemic, reproductive and developmental toxicity of 1-hexene when administered orally, by gavage, to male and female rats using a modified OECD 421 protocol. 1-Hexene was administered at doses of 100, 500, and 1000 mg/kg/day in corn oil; the control group received the vehicle at an equivalent volume. The males were treated for 28 days prior to mating and until euthanasia (44 days of dosing). The females were treated for 14 days prior to mating and during mating, gestation, and lactation until euthanasia (41-55 total days of dosing). Females were allowed to deliver and rear their offspring until lactation day 4. The parental rats were subject to a gross and microscopic examination. Viability and development of the pups were followed through lactation day 4. There was no mortality, and there were no clinical signs of toxicity or differences in body weights, weight gain, feed consumption or organ weights. Copulation and fertility indices, precoital intervals, gestation lengths and pregnancy rates were comparable among the groups, and no signs of prolonged delivery or unusual nesting behaviors were noted. Pup viability, body weights, external observations and necropsy data were comparable among the groups. Pitted kidneys were observed at necropsy for two parental males in the 500 mg/kg/day group and three males in the 1000 mg/kg/day group. Microscopic changes in the kidneys of some male rats from the 100, 500, and 1000 mg/kg/day groups consisted of dose-related accumulations of hyaline droplets in the epithelial cells of the proximal convoluted tubules of the kidneys. In summary, the only treatment-related effect noted in this study was hydrocarbon nephropathy in male rats, which is not considered relevant for human health. The NOAEL for systemic and reproductive toxicity was 1000 mg/kg/day, excluding the finding of male rat hydrocarbon nephropathy. PMID- 10826100 TI - Effect of cocaine on mitochondrial electron transport chain evaluated in primary cultures of neonatal rat myocardial cells and in isolated mitochondrial preparations. AB - Cardiotoxicity is commonly associated with cocaine abuse. Previous studies have indicated that cocaine alters myocardial mitochondrial function. This study was undertaken to investigate the effect of cocaine on activity of the mitochondrial electron transport chain in cultures of neonatal rat cardiomyocytes and in isolated myocardial mitochondria. Cocaine concentrations (10(-5) to 10(-3) M) were used, and these concentrations have been reported in human cocaine users and are within a similar range of cocaine concentrations used in studies in vivo. After 24 hr treatment of cocaine, there was a slight increase in lactate dehydrogenase leakage in the cells treated with cocaine (10(-3) M). Reduction of tetrazolium compounds, neotetrazolium chloride (NTC) and triphenyltetrazolium chloride (TTC) was analyzed in intact cells to assess activity of the mitochondrial electron transport chain. Cocaine (10(-3) M) did not significantly change TTC and NTC reduction. In isolated mitochondria, cocaine (10(-3) M) significantly inhibited glutamate/malate-mediated respiration. These data suggest that cocaine at high concentrations may inhibit complex I of the mitochondrial electron transport chain of myocardial cells. PMID- 10826101 TI - Humoral and cell mediated immune response to cadmium in mice. AB - The effect of 30, 100 and 300 ppm of cadmium chloride (CdCl2) exposure for 35 days on humoral and cell mediated immune response was examined in Swiss Albino mice. Body burden of cadmium in kidney, spleen and liver was determined and histopathology of these organs was also done. Cadmium chloride in doses of 100 and 300 ppm when fed in drinking water caused significant decrease in IgM and IgG titre against sheep red blood cells (SRBC) and a significant decrease in IgG titre against bovine serum albumin (BSA). The delayed type hypersensitivity response to SRBC and splenic T cell proliferation to BSA was also significantly decreased following 100 amd 300 ppm cadmium exposure. Cadmium accumulation in the spleen, liver and kidney was associated with degeneration and inflammatory changes. It is concluded that cadmium causes significant suppression of humoral and cell mediated immune response in mice which could be due to its cytotoxic action on liver, kidney and immune cells. PMID- 10826102 TI - The effect of trifluoperazine on the genotoxicity of bleomycin in cultured human lymphocytes. AB - The effects of trifluoperazine on the toxicity and mutagenicity of bleomycin were examined in cultured human lymphocytes. Lymphocyte cultures were initiated from three adult healthy non-smoking male volunteers. Cultures were exposed to the drugs for either three or twenty hours prior to cell collection. The toxic and clastogenic effects of the different treatments were represented by the reduction in the mitotic indices and the induction of chromosomal aberrations (CA) respectively. Both TFP and BLM significantly increased CA frequencies and reduced the mitotic indices (MI) following all treatments. The reduction in the mitotic indices and the increase in CA frequencies induced by the combined administration of both BLM and TFP were highly significant (p < or = 0.001), but they were not significantly different from the sum of those induced by the separate treatments with the two drugs. These combined treatments, however, potentiated the odds ratios compared to those of the separate drug treatments. Therefore, though the effect of TFP on the clastogenic and cytotoxic effects of BLM was additive, the observed potentiation of the odds ratios of the combined treatments compared to those of the separate treatments suggested a significant enhancement in the expected chemotherapeutic effects of BLM when administered with TFP. PMID- 10826103 TI - The effects of the ALDH2*1/2, CYP2E1 C1/C2 and C/D genotypes on blood ethanol elimination. AB - The effects of CYP2E1 genotypes on the blood ethanol and acetaldehyde levels were investigated in a pair of Japanese volunteers whose ADH2, ADH3 and ALDH2 genotypes were identical but whose CYP2E1 genotypes were different. In the same way, the effects of ALDH2 and ADH2 on the ethanol elimination kinetics were also studied. The predicting 95% confidence bounds determined on regression analysis of the data suggested that after venous injection of ethanol, the blood ethanol and acetaldehyde concentrations in a volunteer normal homozygous for ALDH2 (ALDH2*1/1) were lower than in a heterozygous one (ALDH2*1/2). Also, the blood ethanol and acetaldehyde concentrations in a volunteer with the c2 and C alleles of CYP2E1 (c1/c2 and C/D) were lower than in one without the c2 and C alleles (c1/c1 and D/D). However, there were no significant differences in the blood ethanol and acetaldehyde concentrations between volunteers with ADH2*1 (ADH2*1/1) and without ADH2*1 (ADH2*1/2). PMID- 10826104 TI - Aflatoxins inhibit prolactin secretion by rat pituitary cells in culture. AB - Acute effects of aflatoxins (AF), and in particular hormonal actions, have not been examined as much as chronic toxicity. Thus, we studied the effects of specific AF on prolactin (PRL) secretion by rat pituitary cells in culture. AFB1 and AFQ1 (1 x 10(-4) M) reduced the stimulating effect of dimethyl sulfoxide on PRL secretion by cultured rat pituitary cells. The mechanism responsible for this action is still unknown, but it did not seem to be a non specific toxic effect, because AFB1, at the same concentration, did not significantly alter cell viability, as indicated by the Trypan blue dye-exclusion test. PMID- 10826105 TI - Effect of amiodarone on the membrane bound enzymes of rat intestine. AB - Amiodarone, a cationic amphiphile known for its clinical efficacy as an antiarrhythmic agent, unfortunately causes serious side effects. The present study was undertaken to investigate its intestinal toxicity, on oral administration, using a Wistar rat model. The relationship of drug dose and duration on intestinal toxicity was investigated. Optimum changes were observed after 21 days of AD administration at a dose of 175 mg/Kg body wt/day and this dosage was used for further studies. Histological studies revealed decreased villi and crypt size and reduction in the cellularity of lamina propria. Marked reduction in the activities of Ca(2+)-ATPase, alkaline phosphatase, disaccharidases and Na+, K(+)-ATPase was observed. The reduction in the uptake of 14C-glucose and 14C-glycine, in vivo, was correlated to the reduction in the activities of these enzymes. The reduction in the activities of the intestinal membrane bound enzymes may be attributed to altered morphology of the villi and crypts. PMID- 10826106 TI - Influence of granulating method on physical and mechanical properties, compression behavior, and compactibility of lactose and microcrystalline cellulose granules. AB - The physical and mechanical properties of lactose (LC) and microcrystalline cellulose (MCC) granules prepared by various granulating methods were determined, and their effects on the compression and strength of the tablets were examined. From the force-displacement curve obtained in a crushing test on a single granule, all LC granules appeared brittle, and MCC granules were somewhat plastically deformable. Inter-granular porosity epsilon inter clearly decreased with greater spherical granule shape for both materials. Decrease in intragranular porosity epsilon intra enhanced the crushing force of a single granule Fg. Agitating granulation brought about the most compactness and hardness of granules. In granule compression tests, the initial slope of Heckel plots K1 appeared closely related to ease of filling voids in a granule bed by the slippage or rolling of granules. The reciprocal of the slope in the succeeding step 1/K2 in compression of MCC granules indicated positive correlation to Fg, while in LC granules, no such obvious relation was evident. 1/K2 differed only slightly among granulating methods. Tensile strength of tablets Tt obtained by compression of various LC granules was low as a whole and was little influenced by granulating method. For MCC granules, which are plastically deformable, tablet strength greatly depended on granulation. Granules prepared by extruding or dry granulation gave strong tablets. Tablets prepared from granules made by the agitating method showed particularly low Tt. From stereomicroscopic observation, the contact area between granule particles in a tablet appeared smaller; this would explain the decrease in inter-granular bond formation. PMID- 10826107 TI - PVP solid dispersions for the controlled release of furosemide from a floating multiple-unit system. AB - The poor bioavailability of orally dosed furosemide (FUR) is due to the presence of a biological window in the upper gastrointestinal tract. The purpose of the present study was to develop and optimize in vitro a multiple-unit floating system with increased gastric residence time for FUR. The incomplete release of FUR from the units, related to its low water solubility, led to the preparation and evaluation of different FUR samples to be incorporated into the units. The complete dose release over the actual intragastric residence time of the system (about 8 hr) was achieved by loading both the core and the membrane forming the units with a 1:5 FUR/polyvinylpyrrolidone (FUR/PVP) solid dispersion. Physicochemical analyses suggested the predominant role of the amorphous state of FUR in producing enhanced drug solubility and dissolution rate, which led to the desired release profile from the floating units. PMID- 10826108 TI - Development of terfenadine-pseudoephedrine double-layer tablet dissolution equivalent to core tablet. AB - The terfenadine-pseudoephedrine dosage form discussed here is the sustained release core tablet composed of outer (fast-release) and inner (sustained release) layers. To develop the double-layer tablet dissolution-equivalent to a core tablet, the fast-release and sustained-release layers were prepared using various disintegrants and polymers, respectively. The layer composed of terfenadine/pseudoephedrine/lactose/cornstarch/sodium bicarbonate/hydroxypropylcellulose (HPC)/sodium lauryl sulfate/microcrystalline cellulose (60/10/90/30/20/1/40/1/293 mg), which gave the fast disintegration time and high dissolved amounts of drugs, was selected as the fast-release layer. The dissolved amounts of pseudoephedrine from sustained-release layers increased more with a smaller ratio of ethylcellulose and hydroxypropylmethylcellulose (HPMC). Dissolution mechanism analysis showed the release of pseudoephedrine was proportional to the square root of time, indicating that drug might be released from the layers by Fickian diffusion. The layer composed of pseudoephedrine/ethylcellulose/HPMC (110/30/155 mg), which had similar dissolution amounts of pseudoephedrine as the inner layer of a core tablet, was selected as the sustained-release layer. Furthermore, the dissolved amounts of drugs from the core and double-layer tablets had deviations of less than 5% against the average dissolved amounts of drugs at each time. There was no significant difference between the dissolved amounts of drugs from these tablets at each time in pH 1.2, 4.0, and 6.8 (P > .05). Our results suggest that this double-layer tablet was a dissolution equivalent to the core tablet. PMID- 10826109 TI - Interaction of antimalarial agent artemisinin with cyclodextrins. AB - To obtain an effective solution of the poorly water soluble antimalarial agent artemisinin, the use of several kinds of cyclodextrins (CDs) as solubilizers was examined. The following CDs were used in this study: alpha-CD, beta-CD, gamma-CD as parent CDs, 2-hydroxypropyl-beta-CD (HP-beta-CD), sulfobutyl ether beta-CD (SBE7-beta-CD), heptakis (2,6-di-O-methyl)-beta-CD (DM-beta-CD), 2,3,6-partially methylated-beta-CD (PM-beta-CD) as modified CDs, and glucosyl-beta-CD (G1-beta CD), and maltosyl-beta-CD (G2-beta-CD) as branched CDs. The solubility curves of artemisinin with CDs can all be classified as type AL. The apparent stability constants for artemisinin-parent CD complexes increased in the order of alpha- < gamma- < or = beta-CD. The constants for artemisinin-beta-CD derivative (and beta CD) complexes increased in the order of G2-beta-CD approximately equal to G1-beta CD approximately equal to PM-beta-CD approximately equal to beta-CD < HP-beta-CD < SBE7-beta-CD < DM-beta-CD. These results suggest that the addition of CDs enables the solubilization of artemisinin. PMID- 10826110 TI - Evaluation of validation of a fully instrumented Huttlin HKC 05-TJ laboratory scale fluidized bed granulator. AB - The instrumentation and validation of a laboratory-scale fluidized bed apparatus is described. For continuous control of the process, the apparatus is instrumented with sensors for temperature, relative humidity (RH), and air velocity. Conditions of inlet air, fluidizing air, product, and exhaust air were determined. The temperature sensors were calibrated at temperatures of 0.0 degree C and 99.9 degrees C. The calibration of the humidity sensors covered the range from 12% RH to 98% RH using saturated electrolyte solutions. The calibration of the anemometer took place in a wind tunnel at defined air velocities. The calibrations led to satisfying results concerning sensitivity and precision. To evaluate the reproducibility of the process, 15 granules were prepared under identical conditions. The influence of the type of pump used for delivering the granulating liquid was investigated. Particle size distribution, bulk density, and tapped density were determined. Granules were tableted on a rotary press at four different compression force levels, followed by determination of tablet properties such as weight, crushing strength, and disintegration time. The apparatus was found to produce granules with good reproducibility concerning the granule and tablet properties. PMID- 10826111 TI - Permeation studies comparing cobra skin with human skin using nicotine transdermal patches. AB - Cobra skin (Naja Naja Khaotia) was used as a barrier for an in vitro permeation study using nicotine. Fluxes of nicotine that permeated from Nicotinell through cobra skin (CS) taken from the head, body, and tail were 233.93 +/- 16.08, 206.87 +/- 19.00, and 211.26 +/- 22.93 micrograms/cm2/hr1/2, respectively (n = 6). This indicated no significant difference (p > .05). Abdominal human epidermis (HE), obtained from cadavers, and the CS provided identical permeation kinetics for nicotine, which can be described by Mt = 4M alpha (Dt/pi L2)1/2. The mean flux of nicotine formulated as an acrylic transdermal patch that permeated through HE was 137.92 +/- 67.79 micrograms/cm2/hr1/2 (4 specimens, n = 12), whereas that through CS was 180.13 +/- 41.05 micrograms/cm2/hr1/2 (4 specimens, n = 15). The ratio of the fluxes of nicotine from formulated patches having three different nicotine contents using CS and HE was 1.22 to 1, respectively, for each of the patches irrespective of nicotine content. The coefficients of variation of the nicotine permeated were 22.79% and 49.15% for CS and HE, respectively, that is, a narrower variation of results was obtained with CS. This indicated that CS could be used for nicotine permeation studies. PMID- 10826112 TI - Water vapor adsorption properties of amorphous cefditoren pivoxil evaluated by adsorption isotherms and microcalorimetry. AB - Water vapor adsorption of ground cefditoren pivoxil was studied. The amount of water adsorbed increased with a decrease in the crystallinity of cefditoren pivoxil. It was found from the microcalorimetric measurements that the differential heat of water vapor adsorption at 1.5% adsorbed water increased with decreasing crystallinity of cefditoren pivoxil, suggesting that hygroscopicity of cefditoren pivoxil was enhanced by grinding. These results indicated that hydrophilic adsorption sites in cefditoren pivoxil increased through the grinding process. The results of infrared (IR) spectra examination suggested that the increment of hydrophilic adsorption sites through the grinding process resulted from the change of the environment of the carbonyl groups in two esters and amide. PMID- 10826113 TI - Comparative study of drug release from pellets coated with HPMC or Surelease. AB - The release of metoclopramide hydrochloride (very water soluble cationic drug) and diclofenac sodium (sparingly soluble anionic drug) from pellets coated with hydroxypropylmethylcellulose (HPMC; water-soluble polymer) or ethylcellulose aqueous dispersion (Surelease; water-insoluble polymer) at different coating loads was investigated. The release rates of either drug decreased as the coating load of HPMC increased, but overall, the release was fast, and the majority of both drugs released in about 1 hr, even at the highest coating load. The drug release mechanism for either drug was not affected by the coating load of HPMC or by the type of drug used, and it was found to be mainly diffusion controlled. Diclofenac sodium released slightly more slowly than metoclopramide hydrochloride from HPMC-coated pellets. This was attributed to the lower water solubility of the former drug. The release rate of either drug decreased greatly as the coating load of Surelease increased. The release of both drugs was sustained over 12 hr as the coating load of Surelease increased, and only about 70% of either drug was released after this period at the highest coating load (20%). The mechanism of release of metoclopramide hydrochloride was independent of coating load, and it was predominantly diffusion controlled. However, the mechanism of diclofenac sodium release was dependent on the coating load of Surelease. At low coating loads, diffusion of drug was facilitated due to the presence of more pores at the surface of the coated pellets; therefore, the rate of dissolution of the drug particles was the rate-limiting step. However, at high coating loads, drug release was mainly diffusion controlled. Despite its lower water solubility, diclofenac sodium released slightly faster than metoclopramide hydrochloride from Surelease-coated pellets at equivalent coating loads. PMID- 10826114 TI - Calibration and LOD/LOQ estimation of a chemiluminescent hybridization assay for residual DNA in recombinant protein drugs expressed in E. coli using a four parameter logistic model. AB - Calibration is the process of fitting a model based on reference data points (x, y), then using the model to estimate an unknown x based on a new measured response, y. In DNA assay, x is the concentration, and y is the measured signal volume. A four-parameter logistic model was used frequently for calibration of immunoassay when the response is optical density for enzyme-linked immunosorbent assay (ELISA) or adjusted radioactivity count for radioimmunoassay (RIA). Here, it is shown that the same model or a linearized version of the curve are equally useful for the calibration of a chemiluminescent hybridization assay for residual DNA in recombinant protein drugs and calculation of performance measures of the assay. PMID- 10826115 TI - Characterization of the physicochemical properties of the micelles of platelet activating factor (C18:0). AB - The purpose of this study was to clarify the physicochemical properties of the micelles of platelet-activating factor (PAF; C18:0). The critical micelle concentration (CMC) of PAF (C18:0) was determined (0.20 microM) using fluorescence techniques. The fluidity and the micropolarity of the PAF (C18:0) micelle were similar to those of the micelle of stearoyl lysophosphatidylcholine. PMID- 10826116 TI - Characterization of polyvinylalcohol microspheres of diclofenac sodium: application of statistical design. AB - Microspheres of polyvinylalcohol (PVA) containing diclofenac sodium were prepared by an emulsion-chemical cross-linking method. A statistical design was used to study the variables that affect the preparation of microspheres and to study the release profile of diclofenac from the microspheres. To account for the drug content, a mass balance study of the process was performed. A high concentration of polyvinylalcohol, a high stirring speed, and a low level of glutaraldehyde were found to be important to obtain spherical and discrete microspheres. The concentration of polyvinylalcohol and the amount of heavy liquid paraffin were found to be critical factors in influencing the t50 value. Almost 98% of the total diclofenac sodium added was accounted for in mass balance studies. PMID- 10826117 TI - Preparation and evaluation of a sustained-release formulation of nifedipine HPMC tablets. AB - A nifedipine (NF) polyethylene glycol (PEG) solid dispersion was prepared. Using this solid dispersion, NF hydroxypropylmethylcellulose (HPMC) matrix tablets were prepared. Both the high-viscosity grade HPMC (Methocel K15M) and low-viscosity grade HPMC (Methocel K100) were applied in the tablets to form the matrix. The dissolution and absorption of NF from the tablet were evaluated as a formulation that had a sustained release over 24 hr. The Hixson-Crowell equation and Higuchi equation were used to investigate the dissolution mechanism, and the erosion and diffusion codependent mechanism was established. Adalat GITS 30 was used as a reference dosage form. Each beagle dog was also administered an intravenous injection to obtain the pharmacokinetics parameters. The Loo-Riegelman method was applied to study the in vitro/in vivo correlation of the tested tablets and Adalat GITS 30, and significant correlation was proved. Absolute bioavailability and comparative bioavailability of the tested tablet were studied. The results indicated that the NF HPMC tablet could be an ideal 24-hr sustained-release formulation. PMID- 10826118 TI - Physical structure characterization of theophylline in some acidic film-forming polymers. AB - The physical structure and drug-polymer interactions of theophylline in Eudragit L100, shellac, polyvinyl acetate phthalate (PVAP), cellulose acetate phthalate (CAP), hydroxypropylmethylcellulose acetate phthalate (HPMCP), and hydroxypropylmethylcellulose (HPMC) were studied. The drug-polymer films were prepared by casting and were characterized using powder X-ray diffractometry (PXRD), nuclear magnetic resonance (NMR) spectroscopy, and thin-layer chromatography (TLC). Theophylline was found to recrystallize in the modification II form in all kinds of polymers, which was the same as that recrystallized solely from the solvent system and the original powder. The PXRD and NMR results indicated a superficial drug-polymer interaction between theophylline and Eudragit L100, while there was no evidence of interaction for the others. No drug decomposition was observed by TLC for all drug-polymer mixtures. PMID- 10826119 TI - Ranitidine HCl: tablet film coating acidity and discoloration. AB - The presence of citric acid in the film coating of ranitidine hydrochloride tablets decreased discoloration. PMID- 10826120 TI - Rehabilitation in the community. AB - The Project applied rehabilitation in community (RIC) in Zagreb, Croatia as a model of care with the aim of maximizing physical and mental abilities in a persons' own life environment as well as social integration. From September 96 to June 98, 785 patients with primary physical disablement were admitted: 80% with musculoskeletal conditions, and 20% with traumatic brain or spinal injury, stroke, multiple sclerosis, amputation or rheumatoid arthritis. They were residents of Trnje community (55%), Zagreb area (40%) and Croatia area (5%). Mean age was 44 years (men 39, women 48, range 7-76); 52% of patients were women and 17% were recent war wounded. The RIC programme focused on training for physical condition, activities of daily living, preservation of energy and instructions on self-rehabilitation measures, application of aids, and psychosocial consultation. Patients' families were actively involved. Educational activities involved 1100 participants and RIC was promoted by 182 health and social care agencies, humanitarian organizations, the Church, Red Cross, the University and public. The Project demonstrated that primary health care nurse, social worker, and peer counsellor joined to rehabilitation professionals were able to develop interdisciplinary teamwork and to confirm their roles as CBR workers. PMID- 10826121 TI - Exercise-based rehabilitation for injured workers: programme efficacy and identification of factors predicting programme completion and outcome. AB - A sample of 355 injured workers presenting to a tertiary referral agency for supervised physical activity programmes were assessed for programme completion and changes in work status at the conclusion of the programme. Seventy-five percent of the sample comprised long-term (i.e. greater than six months) injured workers, of whom 45% were unemployed at the commencement of the programme. There was a 15% dropout rate and 49% of completers improved their work status at the end of the programme. A series of stepwise logistic regression analyses were conducted to identify predictors of dropout and improved work status. Joint pathology was the only predictor of dropout. Preprogramme work status, referral source, intervertebral pathology, and time between injury and commencement of the programme predicted post-programme work status. Those workers who were employed, referred by sources other than rehabilitation providers, such as doctors, employers or insurers, did not have a diagnosis including intervertebral pathology and who began their programmes within six months of injury were more likely to improve their work status at the conclusion of the programme. Other factors including age, gender, occupation, insurer category, location of injury (i.e. lumbar or other), and neurological signs predicted neither programme completion nor change in work status. PMID- 10826122 TI - Patient and spouse perceptions of stroke and its rehabilitation. AB - The object of the study was to determine, in stroke patients and their spouses, changes in their perceptions of stroke, information received about it, its management, and its rehabilitation. There were assessments on acute admission, on admission to and discharge from rehabilitation, and six and twelve months after discharge. Participants were 60 twelve month stroke survivors who had undergone an inpatient rehabilitation programme, and 46 spouses. Their perceptions of stroke, of medical information received, of the management of stroke, and of rehabilitation were assessed in a structured interview. Stroke was predominantly perceived by participants in terms of physiological causes and disabling consequences. Its causes were poorly understood, as was the role of doctors and hospitals for people with strokes. The role of rehabilitation was better understood. Many participants felt they were told little about the illness during care. Stroke patients and their spouses adapt with varying success to their changed circumstances. This adaptation will benefit from better communication between medical staff and families in the acute and rehabilitation settings. PMID- 10826123 TI - The development and psychometric assessment of the Turkish version of the Nottingham Health Profile. AB - The Nottingham Health Profile (NHP) is a widely used measure of perceived health status. The aim of the present study was to adapt the NHP for use in Turkey and to test its psychometric properties. Following translation and testing for its face and content validity, 50 patients with osteoarthritis were interviewed on two occasions. Each interview included administration of the NHP and the Stanford Health Assessment Questionnaire (HAQ), a measure of functional disability. Test retest reliability of the new version was satisfactory and comparable with other available language versions. As predicted, high correlations were found between the HAQ and the physical mobility, pain and energy level sections of the NHP and low correlations between the HAQ and emotional reactions, social isolation and sleep, confirming the construct validity of the NHP. It is concluded that the adaptation of the NHP into Turkish was successful but that additional studies are required to assess its suitability for use with other patient populations and its equivalence to other language versions of the measure. PMID- 10826124 TI - Attitudes to disability of Russian occupational therapy and nursing students. AB - Positive student attitudes toward disability are essential to client-centred rehabilitation. Instilling such attitudes in students enrolled in the first occupational therapy (OT) and post-diploma nursing management (NM) courses in Russia was a key objective of the curriculum development team. The team's aim was to combat the traditional Russian attitude of pity, dependence and marginalization. To this end, the introductory classes were developed and taught by teachers from the All Russia Society of Disabled (ARSD). This paper examines the attitudes to disability in Russia and compares the attitudes of three groups of students. Students in the OT and NM programmes (n = 18) in Volgograd were compared to other nursing students in the same college (n = 21) and to nursing students in a Moscow medical school (n = 20). All completed the Scale of Attitudes Toward Disabled Persons (SADP). The Kruskal-Wallis and Mann-Whitney U statistics revealed that students in the OT and NM programmes had significantly more positive scores (P = 0.01) than either of the other groups. This suggests that the strategies adopted by the curriculum team were successful in developing positive attitudes in students. PMID- 10826125 TI - The association between functional status and the number of areas in the body with musculoskeletal symptoms. PMID- 10826127 TI - Local concepts and beliefs of disability: international expert meeting and symposium, Bonn, May 1998. PMID- 10826126 TI - Chronic psychiatric patients: an assessment of treatment and rehabilitation related needs. PMID- 10826128 TI - Modeling of hepatic elimination and organ distribution kinetics with the extended convection-dispersion model. AB - The conventional convection-dispersion (also called axial dispersion) model is widely used to interrelate hepatic availability (F) and clearance (Cl) with the morphology and physiology of the liver and to predict effects such as changes in liver blood flow on F and Cl. An extended form of the convection-dispersion model has been developed to adequately describe the outflow concentration-time profiles for vascular markers at both short and long times after bolus injections into perfused livers. The model, based on flux concentration and a convolution of catheters and large vessels, assumes that solute elimination in hepatocytes follows either fast distribution into or radial diffusion in hepatocytes. The model includes a secondary vascular compartment, postulated to be interconnecting sinusoids. Analysis of the mean hepatic transit time (MTT) and normalized variance (CV2) of solutes with extraction showed that the discrepancy between the predictions of MTT and CV2 for the extended and unweighted conventional convection-dispersion models decreases as hepatic extraction increases. A correspondence of more than 95% in F and Cl exists for all solute extractions. In addition, the analysis showed that the outflow concentration-time profiles for both the extended and conventional models are essentially identical irrespective of the magnitude of rate constants representing permeability, volume, and clearance parameters, providing that there is significant hepatic extraction. In conclusion, the application of a newly developed extended convection-dispersion model has shown that the unweighted conventional convection-dispersion model can be used to describe the disposition of extracted solutes and, in particular, to estimate hepatic availability and clearance in both experimental and clinical situations. PMID- 10826129 TI - The anomalous pharmacokinetics of amiodarone explained by nonexponential tissue trapping. AB - Conventional pharmacokinetic (PK) concepts fail to describe the long-term pharmacokinetics of the extremely cationic amphiphilic drug amiodarone. A nonclassical model based on the phenomenon of trapping at tissue binding sites with very long release times is presented, which implies that a volume of distribution and a steady-state level cannot be defined. In agreement with clinical PK data available in the literature, the model well describes not only single-dose disposition curves but also the persistently increasing plasma concentration-time curve during long-term treatment (up to 5 years) and the washout curve following cessation of therapy. The novel aspect is a long-tailed tissue residence time distribution which is incorporated into a recirculatory model leaving the initial distribution process and the clearance concept unchanged. The underlying theoretical approach, which is known as "strange or anomalous" kinetics in physical sciences, and the fractal scaling property of the model may enhance our understanding of the PK of extremely hydrophobic xenobiotics. PMID- 10826130 TI - Population pharmacokinetics and pharmacodynamics of the anti-CD11a antibody hu1124 in human subjects with psoriasis. AB - The pharmacokinetics of hu1124, a human anti-CD11a antibody, were investigated in human subjects with psoriasis. CD11a is a subunit of LFA-1, a cell surface molecule involved in T cell mediated immune responses. Subjects received a single dose of 0.03, 0.1, 0.3, 0.6, 1, 2, 3, or 10 mg/kg of hu1124 intravenously over 1 3 hr. Blood samples were collected at selected times from 60 min to 72 days after administration. Plasma samples were assayed for hu1124 by ELISA, and pharmacokinetic analyses were performed on the drug plasma concentrations. As the dose of hu1124 was increased, the clearance decreased from 322 ml/day per kg at 0.1 mg/kg to 6.6 ml/day per kg at 10 mg/kg of hu1124. The plasma hu1124 concentration-time profile suggested that the clearance of hu1124 was saturable above 10 micrograms/ml. In addition, treatment with hu1124 caused a rapid reduction in the level of CD11a expression on CD3-positive lymphocytes (T cells) to about 25% of pretreatment levels. Regardless of the hu1124 dose administered, cell surface CD11a remained at this reduced level as long as hu1124 was detectable (> 0.025 microgram/ml) in the plasma. When hu1124 levels fell below 3 micrograms/ml, the drug was rapidly cleared from the circulation and expression of CD11a returned to normal within 7-10 days thereafter. In vitro, half-maximal binding of hu1124 to lymphocytes was achieved at about 0.1 microgram/ml and saturation required more than 10 micrograms/ml. One of the receptor-mediated pharmacokinetic/pharmacodynamic models which was developed describes the dynamic interaction of hu1124 binding to CD11a, resulting in the removal of hu1124 from the circulation and reduction of cell surface CD11a. The model accounts for the continually changing number of CD11a molecules available for removing hu1124 from the circulation based on prior exposure of cells expressing CD11a to hu1124. In addition, the model also accounts for saturation of CD11a molecules by hu1124 at drug concentrations of approximately 10 micrograms/ml, thereby reducing the clearance rate of hu1124 with increasing dose. PMID- 10826131 TI - Absolute bioavailability and absorption profile of cyanamide in man. AB - A pharmacokinetic study of cyanamide, an inhibitor of aldehyde dehydrogenase (EC1.2.1.3) used as an adjuvant in the aversive therapy of chronic alcoholism, has been carried out in healthy male volunteers following intravenous and oral administration. Cyanamide plasma levels were determined by a sensitive HPLC assay, specific for cyanamide. After intravenous administration cyanamide displayed a disposition profile according to a two-compartmental open model. Elimination half-life and total plasma clearance values ranged from 42.2 to 61.3 min and from 0.0123 to 0.0190 L.kg-1.min-1, respectively. After oral administration of 0.3, 1.0, and 1.5 mg/kg x +/- SEM values of Cmax, tmax (median) and AUC were 0.18 +/- 0.03, 0.91 +/- 0.11, and 1.65 +/- 0.27 micrograms.ml-1; 13.5, 13.5, and 12 min; and 8.59 +/- 1.32, 45.39 +/- 1.62, and 77.86 +/- 17.49 micrograms.ml-1.min, respectively. Absorption was not complete and the oral bioavailability, 45.55 +/- 9.22, 70.12 +/- 4.73, and 80.78 +/- 8.19% for the 0.3, 1.0, and 1.5 mg/kg doses, respectively, increased with the dose administered. The models that consider a first-order absorption process alone (whether with a fixed or variable bioavailability value as a function of dose) or with loss of drug due to presystemic metabolism (with zero-order or Michaelis-Menten kinetics) were simultaneously fitted to plasma level data obtained following 1 mg/kg i.v. and 0.3, 1.0, and 1.5 mg/kg oral administrations. The model that best fit the data was that with a first-order absorption process plus a loss by presystemic metabolism with Michaelis-Menten kinetics, suggesting the presence of a saturable first-pass effect. PMID- 10826132 TI - Variability of the model-independent AUC: the one sample per individual case. AB - A theory is developed for estimation of a population value of AUC along with its standard deviation, in the case, when only one concentration-time (C-t) sample is available for each individual. This theory is based on model-independent pharmacokinetics. Integration methods are classified due to their applicability to the presented approach. The main goal of this work is to establish a statistical hypothesis-testing procedure which would make single C-t samples usable for bioequivalence studies. An application of the theory to a number of integration methods currently in use is analyzed in detail. A real data illustration is included. PMID- 10826133 TI - Medical errors hit home. PMID- 10826134 TI - Enclosed automobiles and the risk of hyperthermia. PMID- 10826135 TI - Evaluating and managing intimate partner violence. AB - An estimated 18% to 25% of female primary care and prenatal patients suffer from intimate partner violence, and 31% to 44% of women report some abuse during their lifetimes. Violence is often unreported because women fear the batterer's reprisals, time-consuming court procedures, shame, blame, and unreliable legal sanctions. Screening for domestic violence should be a routine aspect of health care. Clinicians must ask direct questions about violence and detect subtle and overt clues. This article presents the historical context of violence and examines screening, evaluation, treatment, and ethics. PMID- 10826136 TI - The cycle of domestic violence and the barriers to treatment. PMID- 10826137 TI - Understanding the reimbursement process. AB - Nurse practitioners must be fiscally responsible to obtain reimbursement for services provided. A thorough knowledge of the reimbursement process is required to accomplish this goal. This article explores the reimbursement process, generation of revenue, and personal productivity determination for nurse practitioners. Parameters for billing using the resource-based relative value scale, the Evaluation and Management Documentation Guidelines, the International Classification of Disease, and the Physicians' Current Procedural Terminology code book are explained. PMID- 10826138 TI - Rotator cuff pathology and subacromial impingement. AB - Musculoskeletal complaints are one of the most common reasons for primary care office visits, and rotator cuff disorders are the most common source of shoulder pain. Subacromial impingement with subsequent tendinitis and bursitis is frequently found in young adult patients. Rotator cuff tears are a common cause of shoulder pain in patients over age 40. The majority of subacromial impingement and incomplete rotator cuff tears may be successfully managed with conservative treatment. This article discusses anatomic function of the glenohumeral joint and subacromial space, etiology of subacromial impingement and rotator cuff disorders, examination of the shoulder, diagnostic testing, and treatment of subacromial impingement and rotator cuff disorders in the primary care setting. PMID- 10826139 TI - Understanding Klinefelter's syndrome. PMID- 10826140 TI - Never too late to quit. PMID- 10826141 TI - The preceptor/student relationship: implications for practicum evaluation. PMID- 10826142 TI - Building a safety program from scratch. PMID- 10826143 TI - "Use in a well ventilated area". PMID- 10826144 TI - Reducing the hazards of high heat. PMID- 10826145 TI - Healthy hydration. PMID- 10826146 TI - CO2 measurements for IAQ analysis. AB - These examples illustrate the utility of carbon dioxide measurements as a diagnostic tool for indoor air quality. Carbon dioxide is sometimes said to be an unreliable indicator because of uneven mixing. However, even though mixing does occur slowly in very still air, in nearly all cases there is enough air movement that mixing is not a misleading factor for diagnostic purposes. Measurements do have to be recorded with enough context for proper interpretation. For example, to distinguish between the effects of natural and mechanical ventilation. Or to understand how the results are affected by the number of people in the building and what they are doing. Or how they relate to the nature and layout of the HVAC system and how it is adjusted or intended to be used. Carbon dioxide measurements say little about how healthy an indoor environment is. But they can tell a great deal about how comfortable it is and, with good observation, can give important clues as to where to look for factors that may be affecting health. The interaction of subjective and objective elements in the interpretation of CO2 measurements makes them a fascinating and powerful tool in the hands of a careful investigator. PMID- 10826147 TI - A fixed monitoring solution. AB - A comprehensive, plant-wide hazardous gas risk management system needs to be capable of bringing together all of the necessary information that allows the operator to make decisions correctly and quickly that protect plant and personnel. Such a system incorporates digital communication links from each sensor to the controller and links the gas detection control system to Ethernet for plant-wide access. The digital communications link between sensors and controller allows the use of modules for alarm relays and displays to be easily added wherever they are needed. Digital communications from the controller allow this valuable information to be used with MMI programs or DCS. PMID- 10826148 TI - The basic IH guide to monitoring ventilation. PMID- 10826149 TI - 'Enviromachining' cuts coolant. PMID- 10826150 TI - Service life software for organic vapor cartridges. PMID- 10826151 TI - Eyewash standards & guidelines for the workplace. PMID- 10826152 TI - Protective coveralls: reborn as a performance product. PMID- 10826153 TI - Full compliance. PMID- 10826155 TI - Promoting good research practice. AB - Embarking on a new millennium provides the stimulus both to take stock and also to look forward. In the field of medical statistics there is much to make us feel proud and excited. Rapid methodological developments together with parallel developments in computer technology have enormously expanded our statistical repertoire. At the same time, the high profile attained by the evidence-based medicine movement means that the importance of our discipline is recognized more widely then ever before. However, any medical statistician who is involved in medical publishing, or who is even a regular reader of the medical literature, must be aware of the yawning chasm between what is recognized as good statistical practice and what is actually published. Poor study design, inappropriate analysis and selective reporting are commonplace. In my opinion the most important challenge currently facing our profession is the task of bridging this chasm. PMID- 10826154 TI - Biostatistics in the new millennium: a consulting statistician's perspective. AB - This article attempts to predict the future for biostatistics and biostatisticians in the twenty-first century. Life will certainly be more complex and there will be growth in population, large corporations, and globalization generally. However, there will continue to be problems relating to biology, medicine, health and the environment, so biostatisticians can surely play an important role, if they are willing to adapt to changing circumstances. Personal views are expressed concerning those areas of most recent development that seem likely to be continued in the coming years: applications (clinical trials, epidemiology, vital statistics), philosophies, models, advances in computing, and the profession of biostatistics. PMID- 10826156 TI - The next 10 years of biostatistics. AB - I return to an article on the future of statistics written by Sir Maurice Kendall some 30 years ago. I draw some parallels between the state of general statistics at that time and the state of biostatistics today, and try to learn from the successes and omissions in Sir Maurice's predictions to make my own limited projections for the next 10 years. PMID- 10826157 TI - Medical statistics--no time for complacency. AB - Statistical science plays an important role in medical research. Whether this will continue to be so, depends to a large extent on the future of fields like clinical trials and epidemiology. Will these continue to play the important role they do today in the development of medicine? Traditional, frequentist, methodology still dominates in medical statistics. Statisticians should have an open and pragmatic attitude towards new approaches. Survival analysis is a field of biostatistics that has been through considerable recent development. The need for further methodological development in this field is stressed. There is also need for connecting statistical work more intimately to biological knowledge than is the case today. PMID- 10826158 TI - Statistical methods in public health and epidemiology: a look at the recent past and projections for the next decade. AB - This article attempts to prognosticate from past patterns, the type of statistical methods that will be used in published public health and epidemiological studies in the decade that follows the millennium. With this in mind, we conducted a study that would characterize trends in use of statistical methods in two major public health journals: the American Journal of Public Health, and the American Journal of Epidemiology. We took a probability sample of 348 articles published in these journals between 1970 and 1998. For each article sampled, we abstracted information on the design of the study and the types of statistical methods used in the article. Our major findings are that the proportion of articles using statistical methods as well as the mean number of statistical methods used per article has increased dramatically over the three decades surveyed. Also, the proportion of published articles using study designs that we classified as analytic has increased over the years. We also examined patterns of use in these journals of three statistical methodologies: logistic regression, proportional hazards regression, and methods for analysis of data from complex sample surveys. These methods were selected because they had been introduced initially in the late 1960s or early 1970s and had made considerable impact on data analysis in the biomedical sciences in the 1970s-90s. Estimated usage of each of these techniques remained relatively low until user-friendly software became available. Our overall conclusions are that new statistical methods are developed on the basis of need, disseminated to potential users over a course of many years, and often do not reach maximum use until tools for their comfortable use are made readily available to potential users. Based on these conclusions, we identify certain needs that are not now being met and which are likely to generate new statistical methodologies that we will see in the next decade. PMID- 10826159 TI - Exponential family models and statistical genetics. AB - This article describes the evolution of applied exponential family models, starting at 1972, the year of publication of the seminal papers on generalized linear models and on Cox regression, and leading to multivariate (i) marginal models and inference based on estimating equations and (ii) random effects models and Bayesian simulation-based posterior inference. By referring to recent work in genetic epidemiology, on semiparametric methods for linkage analysis and on transmission/disequilibrium tests for haplotype transmission this paper illustrates the potential for the recent advances in applied probability and statistics to contribute to new and unified tools for statistical genetics. Finally, it is emphasized that there is a need for well-defined postgraduate education paths in medical statistics in the year 2000 and thereafter. PMID- 10826160 TI - Simulating the impact of cross resistance between Bt toxins in transformed clover and apples in New Zealand. AB - Simulation were conducted to guide development of resistance management strategies aimed at prolonging the usable life of B. thuringiensis (Bt) endotoxins in multiple cropping situations, where different crops expressing Bt endotoxins are host plants for a common pest. We used the New Zealand apple and clover model ecosystem to explore the relative impact on the rate of resistance development of varying levels of cross-resistance between different toxins expressed in these 2 potentially Bt-transformed crops. These 2 crops are hosts for a complex of leaf-rollers in New Zealand, including the light-brown apple moth, used here as the model pest. Cross-resistance was varied between 0.0 and 0.5 (zero to partial cross-resistance) to allow for the case in which selection by one plant has a potential effect on resistance to the toxin in another plant. The largest factor affecting the evolution of resistance was the total habitat area occupied by transgenic orchards. The proportion of the clover habitat that was transformed was also an important factor, even in the absence of cross resistance. The effect of increasing the proportion of the second transformed crop (clover) acted on resistance evolution mainly by reducing the external refuge of susceptibility for the transgenic orchards. Hence, the ecological implications of reducing the available source of susceptible insects from clover, which can help to slow resistance development in the orchard ecosystem, had a more significant impact than the presence of cross-resistance. Partial cross resistance between different toxins in the separate crops was overall of relatively minor importance. These simulations have implications for deployment decisions for individual transformed crops in multiple cropping systems, where there is the potential for the crops to serve as refugees for each other. These decisions may need to focus less on cross-resistance between toxins, than on economic trade-offs between the relative roles of individual crops as refugia maintaining susceptibility in the system as a whole. PMID- 10826161 TI - "Inert" formulation ingredients with activity: toxicity of trisiloxane surfactant solutions to twospotted spider mites (Acari: Tetranychidae). AB - Organosilicone molecules are important surfactant ingredients used in formulating pesticides. These methylated silicones are considered inert ingredients, but their superior surfactant properties allow them to wet, and either suffocate or disrupt important physiological processes in mites and insects. Aqueous solutions of the tri-siloxane surfactants Silwet L-77, Silwet 408, and Silwet 806 were bioassayed against adult female two-spotted spider mites, Tetranychus urticae Koch, with leaf dip methods to compare their toxicity with organosilicone molecules containing bulkier hydrophobic components. All three tri-siloxanes in aqueous solutions were equivalently toxic (LC50 = 5.5-8.9 ppm), whereas Silwet L 7607 solutions were less toxic (LC50 = 4,800 ppm) and Silwet L-7200 was nontoxic to mites. In another experiment, the toxicity of Silwet L-77 was affected by the wettability of leaf surfaces. The LC50 shifted from 22 to 84 ppm when mites were tested on bean and strawberry leaf disks, respectively. Droplet spreading on paraffin and surface tension were both related to the toxicity of surfactant solutions. Surface tensions of solutions below 23 mN/m caused > 90% mite mortality in leaf dip bioassays. A field test of Conserve SC and its formulation blank, with and without Dyne-Amic adjuvant (a vegetable oil-organosilicone surfactant mixture) revealed that Dyne-Amic had the greatest miticidal contribution, reducing mite populations by 70%, followed by formulation inactive ingredients. Spinosad, the listed active ingredient in Conserve, only contributed miticidal activity when synergized by Dyne-Amic. Researchers should include appropriate surfactant or formulation blank controls when testing insecticides or miticides, especially when using high spray volumes. PMID- 10826162 TI - Laboratory evaluation of miticides to control Varroa jacobsoni (Acari: Varroidae), a honey bee (Hymenoptera: Apidae) parasite. AB - A laboratory bioassay was developed to evaluate miticides to control Varroa jacobsoni (Oudemans), an important parasite of the honey bee, Apis mellifera L. Bees and mites were exposed to applications of essential oil constituents in petri dishes (60 by 20 mm). The registered mite control agents tau-fluvalinate (Apistan) and formic acid also were evaluated as positive controls. Treatments that caused high mite mortality (> 70%) at doses that produced low bee mortality (< 30%) were considered mite selective. The six most selective of the 22 treatments tested (clove oil, benzyl acetate, thymol, carvacrol, methyl salicylate, and Magic3) were further evaluated to estimate LD50 values and selectivity ratios (A. mellifera LD50/V. jacobsoni LD50) at 24, 43, and 67 h after exposure. Tau-fluvalinate was the most selective treatment, but thymol, clove oil, Magic3, and methyl salicylate demonstrated selectivity equal to or greater than formic acid. The effect of mode of application (complete exposure versus vapor only) on bee and mite mortality was assessed for thymol, clove oil, and Magic3 by using a 2-chambered dish design. Estimated V. jacobsoni LD50 values were significantly lower for complete exposure applications of thymol and Magic3, suggesting that both vapor and topical exposure influenced mite mortality, whereas estimated values for clove oil suggested that topical exposure had little or no influence on mite mortality. These results indicate that essential oil constituents alone may not be selective enough to control Varroa under all conditions, but could be a useful component of an integrated pest management approach to parasitic mite management in honey bee colonies. PMID- 10826163 TI - Comparative laboratory toxicity of neem pesticides to honey bees (Hymenoptera: Apidae), their mite parasites Varroa jacobsoni (Acari: Varroidae) and Acarapis woodi (Acari: Tarsonemidae), and brood pathogens Paenibacillus larvae and Ascophaera apis. AB - Laboratory bioassays were conducted to evaluate neem oil and neem extract for the management of key honey bee (Apis mellifera L.) pests. Neem pesticides inhibited the growth of Paenibacillus larvae (Ash, Priest & Collins) in vitro but had no effect on the growth of Ascophaera apis (Olive & Spiltoir). Azadirachtin-rich extract (neem-aza) was 10 times more potent than crude neem oil (neem oil) against P. larvae suggesting that azadirachtin is a main antibiotic component in neem. Neem-aza, however, was ineffective at controlling the honey bee mite parasites Varroa jacobsoni (Ouduemans) and Acarapis woodi (Rennie). Honey bees also were deterred from feeding on sucrose syrup containing > 0.01 mg/ml of neem aza. However, neem oil applied topically to infested bees in the laboratory proved highly effective against both mite species. Approximately 50-90% V. jacobsoni mortality was observed 48 h after treatment with associated bee mortality lower than 10%. Although topically applied neem oil did not result in direct A. woodi mortality, it offered significant protection of bees from infestation by A. woodi. Other vegetable and petroleum-based oils also offered selective control of honey bee mites, suggesting neem oil has both a physical and a toxicological mode of action. Although oils are not as selective as the V. jacobsoni acaricide tau-fluvalinate, they nonetheless hold promise for the simultaneous management of several honey bee pests. PMID- 10826164 TI - Sunlight persistence and rainfastness of spray-dried formulations of baculovirus isolated from Anagrapha falcifera (Lepidoptera: Noctuidae). AB - Nuclear polyhedrosis viruses such as the one isolated from the celery looper, Anagrapha falcifera (Kirby) (AfMNPV), have the potential to be successful bioinsecticides if improved formulations can prevent rapid loss of insecticidal activity from environmental conditions such as sunlight and rainfall. We tested 16 spray-dried formulations of AfMNPV to determine the effect of different ingredients (e.g., lignin, corn flour, and so on) on insecticidal activity after simulated rain and simulated sunlight (at Peoria, IL) and natural sunlight exposures (at Tifton, GA). The most effective formulation contained pregelatinized corn flour and potassium lignate, which retained more than half of its original activity after 5 cm of simulated rain, and almost full activity after 8 h of simulated sunlight. In Georgia, formulations made with and without lignin were compared for persistence of insecticidal activity when exposed to natural sunlight. In addition, the effect of fluorescent brighteners as formulation components and spray tank additives was tested. Results showed that the formulations with lignin had more insecticidal activity remaining after sunlight exposure than formulations without lignin. The inclusion of brighteners in the formulation did not improve initial activity or virus persistence. However, a 1% tank mix significantly enhanced activity and improved persistence. Scanning electron micrographs revealed discreet particles, and transmission electron micrographs showed virus embedded within microgranules. Results demonstrated that formulations made with natural ingredients could improve persistence of virus-based biopesticides. PMID- 10826165 TI - Assessment of microencapsulated formulations for improved residual activity of Bacillus thuringiensis. AB - Bacillus thuringiensis Berliner is a highly efficacious bioinsecticide used to control lepidopteran pests in the field. Unfortunately, it has limited residual activity on plants because sunlight inactivates spores and crystals and they can be washed off by rain. To minimize loss of activity, formulations must contain UV protectants, stickers, or both. We tested approximately 80 formulations and determined optimal combinations of ingredients and spray drying conditions for improving B. thuringiensis residual activity after simulated rain and simulated sunlight. B. thuringiensis stability, after simulated sunlight (xenon light/8 h) and rain (5 cm/50 min), was improved using formulations based on lignin, corn flours, or both, with up to 20% of the active ingredient, when compared with technical powder or Dipel 2x in laboratory assays. Two formulations, made with corn flours or lignin + pregelatinized corn flour (PCF), killed 51.6 and 75.3% of Ostrinia nubilalis (Hubner) neonates after rain, respectively, versus 27% for technical powder. When the insecticidal activity was tested after simulated sunlight, corn flour-based formulations killed 78.5% of test larvae, and the lignin + PCF formulation killed 70.4%, in contrast to technical powder which caused an average of 29% mortality. Formulations made with Dipel 2x rather than technical powder, caused 62.5% mortality (corn flour-based formulations), and 72.3% mortality (lignin + PCF), versus 53.4% for Dipel 2x after rain. When tested after simulated sunlight, formulations killed 95% of the larvae (average of both formulations) versus 82% for Dipel 2x. In a field test, formulations were applied to cabbage and insecticidal activity was determined against Trichoplusia ni (Hubner) neonates exposed to treated leaves. Insecticidal activity of the corn flour-based formulations was comparable to Dipel 2x for 4 d after treatment, but was significantly better than Dipel 2x 7 d after application. A lignin and PCF based formulation showed significantly higher residual activity than Dipel 2x, 4 and 7 d after application. PMID- 10826166 TI - Identification of ice-nucleating active Pseudomonas fluorescens strains for biological control of overwintering Colorado potato beetles (Coleoptera: Chrysomelidae). AB - Laboratory studies were conducted to identify ice-nucleating active bacterial strains able to elevate the supercooling point, the temperature at which freezing is initiated in body fluids, of Colorado potato beetles, Leptinotarsa decemlineata (Say), and to persist in their gut. Adult beetles fed ice-nucleating active strains of Pseudomonas fluorescens, P. putida, or P. syringae at 10(6) or 10(3) bacterial cells per beetle had significantly elevated supercooling points, from -4.5 to -5.7 degrees C and from -5.2 to -6.6 degrees C, respectively, immediately after ingestion. In contrast, mean supercooling point of untreated control beetles was -9.2 degrees C. When sampled at 2 and 12 wk after ingestion, only beetles fed P. fluorescens F26-4C and 88-335 still had significantly elevated supercooling points, indicating that these strains of bacteria were retained. Furthermore, beetle supercooling points were comparable to those observed immediately after ingestion, suggesting that beetle gut conditions were favorable not only for colonization but also for expression of ice-nucleating activity by these two strains. The results obtained from exposure to a single, low dose of either bacterial strain also show that a minimum amount of inoculum is sufficient for establishment of the bacterium in the gut. Persistence of these bacteria in Colorado potato beetles long after ingestion was also confirmed using a polymerase chain reaction technique that detected ice-nucleating active bacteria by virtue of their ina genes. Application of these ice-nucleating active bacteria to elevate the supercooling point of this freeze-intolerant insect pest could significantly reduce their winter survival, thereby reducing local populations and, consequently, crop damage. PMID- 10826167 TI - Effects of the chitin synthesis inhibitor buprofezin on survival and development of immatures of Chrysoperla rufilabris (Neuroptera: Chrysopidae). AB - Effects of buprofezin (Applaud), a chitin synthesis inhibitor, on survival and development of eggs, three instars, and pupae of Chrysoperla rufilabris (Burmeister) were determined in the laboratory. Buprofezin at three tested concentrations (100, 500, and 1,000 mg [AI]/liter) did not affect the viability and development of eggs when the eggs were treated, or third instars and pupae when those stages were treated. Although the degree of effects by buprofezin on larvae varied with instar, buprofezin at the higher concentrations (500 and 1,000 mg [AI]/liter) reduced survival rates 17-47% and prolonged the overall development from first instars to adult emergence by 2 or 3 d when first instars were treated, indicating that the first instar is the most vulnerable stage. When second instars were treated, the survival of C. rufilabris from second instars to pupae was not significantly affected. However, the developmental time from second instar to adult emergence was longer in the treatments with the highest concentration (1,000 mg [AI]/liter) than that with the lowest concentration (100 mg [AI]/liter). The compatibility of buprofezin with natural enemies in integrated pest management programs is discussed. PMID- 10826168 TI - Phenology and spatial pattern of Typhaea stercorea (Coleoptera: Mycetophagidae) infesting stored grain: estimation by pitfall trapping. AB - The hairy fungus beetle, Typhaea stercorea (L.), occurs frequently in stored grain, often in large numbers. Populations infesting stored barley in Minnesota, corn in South Carolina, and wheat in Florida were sampled by means of grain probe traps. Spatial distribution of the species was examined by contour analysis of trap catch. In South Carolina, corn was sampled at 2 locations over 2 storage seasons, and temperature, moisture content, and malathion residues were measured. These data were used to examine phenology as well as spatial distribution, and showed peak trap catch shortly after harvest in the fall, and in the spring. This pattern followed seasonal changes in grain temperature, but there was no apparent relationship of trap catch to either grain moisture content or malathion residue. The populations of T. stercorea were not distributed randomly, but were largely concentrated in 1 or very few aggregations associated with the "spoutline," a region high in foreign material and broken grain that forms near the center of a bin as it is loaded. However, the spatial patterns were dynamic, even on a very small time scale (week to week). Numbers of insects in aggregations rose and fell, the areas involved expanded and contracted, the centers shifted, and secondary centers appeared and disappeared. These changes were apparently in response to changing patterns of grain temperature and moisture content. Secondary centers of aggregation often formed in warmer grain along bin walls. PMID- 10826169 TI - Effect of nymphal diet on adult predation behavior in Orius majusculus (Heteroptera: Anthocoridae). AB - The predatory bug Orius majusculus (Reuter) was reared on 2 different diets during the nymphal stages. The 1st group was exclusively offered eggs of Ephestia kuehniella Zeller (Lepidoptera: Pyralidae), a standard diet for O. majusculus production. The 2nd group was exclusively offered 4th instars of the pea aphid, Acyrtosiphon pisum (Harris). Subsequently, adult predatory behavior in experimental arenas containing A. pisium was recorded using 2 video cameras. One camera permitted observation of the predator's contact with the prey, where the 2nd camera viewed the arena from above to record the path taken by O. majusculus adults before and after contact with prey. When O. majusculus were reared on aphids, adult bugs successfully located and consumed 55% of experimental prey and continued prey search behavior after each aphid meal. O. majusculus adults that had no experience of aphid predation as nymphs, did not prey on aphids in the experimental arena. The mean walking speed of this group of predators increased from 5.9 +/- 1.2 mm/s to 9.8 +/- 0.7 mm/s after contact or detection of prey, indicating that predators rapidly moved away from unfamiliar prey. Moreover, for egg-reared O. majusculus, all contacts between aphid and predator were lateral, along the side of the prey and were effectively repelled by an aphid kicking response. In contrast, 83% of attacks by aphid-reared O. majusculus were directed at the head or posterior abdomen for which the prey could not defend themselves adequately. When egg-reared O. majusculus were exposed to novel aphid prey for 1 8 d, the frequency of aphid attack increased significantly. We conclude that the standard diet used for rearing O. majusculus may adversely affect the efficiency of this predator as an agent of biological control. PMID- 10826171 TI - Population dynamics of western flower thrips (Thysanoptera: Thripidae) in nectarine orchards in British Columbia. AB - Development of a control strategy for thrips attacking nectarine trees depends on an understanding of their phenology, distribution, and life history as related to characteristics of nectarine orchards. To this end, we compared the overwintering behavior, distribution, and abundance of western flower thrips, Frankliniella occidentalis (Pergande), among 11 nectarine orchards located in the dry central interior of British Columbia, Canada, during 1993 and 1994. Western flower thrips emerged from areas not previously used for agriculture (wild areas) and from within orchards before trees were out of dormancy. Flight of thrips within and around orchards peaked during early bud development, with a second major peak several weeks later after husk fall as the next generation emerged. Orchards protected from wild areas by other orchards had the lowest densities of thrips in buds. Density estimates of western flower thrips on trees were not affected by location of trees within orchards or buds within trees, but most thrips were found in the most developed buds on a tree at any one time. Thrips were not found within buds until petal was first visible on the buds. Larval feeding on buds at early petal fall resulted in serious surface russetting of fruit. PMID- 10826170 TI - Monitoring of the European corn borer (Lepidoptera: Crambidae) in central Maine. AB - Pheromone trap types and within-field trap locations were compared for their effectiveness in monitoring the flight activity of European corn borer, Ostrinia nubilalis (Hubner), and its relationship to egg mass density and crop damage in sweet corn in central Maine from 1995 to 1996. The use of both 3:97 Z:E-11 tetradecenyl acetate and 97:3 Z:E-11 tetradecenyl acetate pheromone blends confirmed that European corn borer in central Maine is attracted to both pheromone lure types. European corn borer moths were captured predominantly with the E-lure type than with the Z-lure type in both years. The Scentry Heliothis trap was more effective than the Multi-Pher trap, but similar to the pheromone baited water pan trap for monitoring European corn borer flights. With the Scentry Heliothis trap, the grassy border and 1st corn rows were the best locations for moth capture during the early flight period, but during the peak flight period, traps located in the middle of the field caught the most moths. Corn damage was recorded before moth captures in some sites and before egg mass counts in others, indicating poor efficacy of traps for early flights. Significant and positive correlations were found between moth captures in the midfield location and egg mass counts, and corn leaf damage, and between egg mass counts and corn leaf damage. However, low coefficients of variation suggest that pheromone trap captures were not good predictors of European corn borer leaf damage in sweet corn. PMID- 10826172 TI - Population dynamics and effects of Oebalus ornatus (Hemiptera: Pentatomidae) on rice yield and quality in southwestern Colombia. AB - Field experiments were conducted from 1989 to 1992 to determine the effects of Oebalus ornatus (Sailer) on Cica 8 rice, Oryza sativa L., yield and to study the population dynamics of the insect. The effect of O. ornatus was measured for seven population levels (0, 2, 3, 4, 8, 12, and 24 sexed pairs per 20 panicles) at three stages of grain development (flowering, milk, and soft dough stage). Insect feeding during the flowering and milk stages of grain development caused more damage than feeding during the soft dough stage. The action threshold was calculated to be 14 O. ornatus adults per square meter for the flowering and milk stages and 67 for the soft dough stage of grain maturity. Population densities that would reduce rice yield in southwestern Colombia were not observed during the 3 yr of the study. PMID- 10826173 TI - Field tests of environmentally friendly malathion replacements to suppress wild Mediterranean fruit fly (Diptera: Tephritidae) populations. AB - This article reports a large-scale field test of two environmentally friendly malathion replacements on wild populations of the Mediterranean fruit fly, Ceratatis capitata (Wiedemann): spinosad, a bacteria-derived toxin, and phloxine B, a red dye with phototoxic properties. The comparison test was conducted on 11 coffee fields infested with wild populations of Mediterranean fruit fly on the Hawaiian island of Kauai with 8-wk protein bait sprays with and without toxicants. To assess effectiveness, adults were trapped and larval infestation levels were evaluated with fruit collections. Malathion was found to be the most effective treatment. However, the two replacements gave significant levels of control, and because they are environmentally safer, should be considered for eradicating incipient populations of this invasive species of fruit fly. Cage tests were also conducted to ensure that the wild flies consumed the bait and to assess how long the bait-toxicant combination remained effective in the field. Although spinosad and phloxine B were found to be effective up to 1 wk, malathion remained effective at least 2 wk. PMID- 10826174 TI - Effects of applaud on the growth of silkworm (Lepidoptera: Bombycidae). AB - An experiment was carried out to evaluate the effect of the insecticide Applaud (buprofezin 25% WP) on the silkworm Bombyx mori (L.). This insecticide belongs to the class of insect growth regulators (IGR). The larvae were fed on leaves treated with 3 different concentrations (0.5, 1, 2 g/liter) of Applaud on the 1st d of each instar. Analysis of data with the Tukey-Kramer test at 1% significant level revealed that mortality and larval duration did not differ among the treatments. On the contrary, the larval weight, which was estimated just before mounting (procedure during which the mature larva climbing on a branch or other material to spin the cocoon), differed among the treatments. Also, cocoon weight, shell weight, and cocoon sericin and fibroin content were different among the treatments, except the shell cocoon ratio. Maximum weight was observed in the controls and minimum in the last instar treatments. Our data suggest that supplementation of Applaud through food to larvae does not affect their mortality rate. On the contrary, it affects larval growth and cocoon parameters. PMID- 10826175 TI - Laboratory and field evaluations of imidacloprid against Melanoplus sanguinipes (Orthoptera: Acrididae) on small grains. AB - The toxicity of imidaloprid to the migratory grasshopper, Melanoplus sanguinipes (F.), was measured in bioassays, greenhouse trials, and field trials. An LD50 of 53 and 86 ppm for the oral/topical applications of imidacloprid confirmed a low toxicity for this chemical when compared with carbofuran as a standard. However, 100% debilitation was observed at concentrations of > or = 1 ppm. Grasshoppers exhibited leg flexing, abdominal quivering, and tremors before becoming motionless and appearing dead. Knockdown was temporary with a high percentage of recovery within 1 h. Efficacy and feeding damage were determined from artificial infestations of M. sanguinipes at the 2nd, 4th, and early tillering growth stages of winter and spring wheat treated with foliar and seed treatments of imidacloprid. All rates of imidacloprid tested resulted in < 45% mortality to 4th instar and adult M. sanguinipes in the greenhouse and field. Although efficacy was low, high rates of debilitation in bioassays suggest that improved control may be gained by combining imidacloprid with insect pathogens or additional chemicals. PMID- 10826176 TI - Lethal and sublethal effects of selected insecticides and an insect growth regulator on the boll weevil (Coleoptera: Curculionidae) ectoparasitoid Catolaccus grandis (Hymenoptera: Pteromalidae). AB - A laboratory culture of Catolaccus grandis (Burks), an ectoparasitoid of the boll weevil, Anthonomus grandis grandis Boheman, was exposed to lethal and sublethal doses of insecticides and an insect growth regulator using a spray chamber bioassay. Materials tested were azinphos-methyl, endosulfan, fipronil, malathion, cyfluthrin, dimethoate, spinosad, methyl parathion, acephate, oxamyl, and tebufenozide. At full rates, spinosad was significantly less toxic to female C. grandis than other treatments except endosulfan. Fipronil and malathion were significantly more toxic to females than other treatments. Most of the chemicals tested were highly toxic to male C. grandis; spinosad was least toxic. At reduced rates, most of 4 selected chemicals tested were low in toxicity to C. grandis; however, a reduced rate of malathion was significantly more toxic to females than other treatments. No C. grandis pupae developed from parasitism during a 24-h treatment period with malathion or spinosad. The sex ratio of progeny from sprayed adults appeared to be unaffected by the treatments. PMID- 10826177 TI - Africanized honey bees (Hymenoptera: Apidae) have a greater fidelity to sunflowers than European bees. AB - A study of sunflower, Helianthus annuus L., pollen collection by Africanized and European honey bees, Apis mellifera L., was conducted in a hybrid seed production field in Argentina. Africanized honey bees collected significantly larger proportions of sunflower pollen than did European honey bees. The result suggests that Africanized bees would be more efficient for commercial sunflower seed production. PMID- 10826178 TI - Action windows for optimum control of rangeland grasshoppers (Orthoptera: Acrididae). AB - Different assemblages of primary and secondary pest grasshopper species were found in 5 different rangeland plant communities in western North Dakota. The action window for their control (i.e., the interval when control is likely to be efficacious) with short-lived insecticides can be visualized in terms of time, grasshopper phenology, or plant phenology. Action windows ranged from 14 d duration in habitats dominated by western wheatgrass to 38 d duration in habitats dominated by needle-and-thread or by crested wheatgrass. Action windows opened 11 15 d later in rhizominous grass habitats than in bunchgrass habitats because of increased presence of late-hatching secondary pest species. In all habitat types, action windows closed primarily in response to imminent oviposition by a ubiquitous primary pest species, Melanoplus sanguinipes (F.). In all habitat types, action windows tended to open during seed development of 2 common and conspicuous grasses, needle-and-thread and green needlegrass. No such consistent indicator for closure of action windows was detected. PMID- 10826179 TI - Effect of crop rotation on populations of Epitrix tuberis (Coleoptera: Chrysomelidae) in potato. AB - The effect of crop rotation on populations of tuber flea beetle, Epitrix tuberis Gentner, in potatoes was investigated using data supplied by an integrated pest management (IPM) company and Geographic Information System software and conventional statistical methods. Using combined 1995 and 1996 data, beetles of the overwintered and F1 generations in both the interior and edges of potato fields showed a significant linear increase with an increase in the preceding consecutive years (0, 1, and 2 years) that the current years' crop was planted to potatoes. Populations were significantly higher in nonrotated fields compared with rotated fields. Both the percentage of the cropping region requiring insecticidal control of tuber flea beetles and the cost of insecticides per hectare of potatoes grown increased linearly with an increase in the number of previous years planted to potatoes. Not practicing crop rotation resulted in a 4.2-7.3% increase in the cropping region requiring insecticidal control of tuber flea beetles. The cost of controlling beetles in potato fields planted to potatoes for 3 consecutive years was up to $20/ha greater than potatoes rotated from the preceding year. Beetle counts from the interior of rotated potato fields never exceeded threshold levels when field edges also were below threshold. It is concluded that sampling of overwintered beetles in interior sites of rotated fields could be abandoned, and only 1 monitoring scout rather than 2 would be necessary to monitor a field during this time. From these results, we concluded that rotating potato crops would reduce spray costs to the farmer and monitoring costs to IPM companies. PMID- 10826180 TI - Relative suitability of crested wheatgrass and other perennial grass hosts for the Russian wheat aphid (Homoptera: Aphididae). AB - The Russian wheat aphid, Diuraphis noxia (Mordvilko) (Homoptera: Aphididae), reproduces parthenogenetically in North America and must survive year-round on host plants, including in late summer when small grains are not in cultivation. During this time, cool-season perennial wheatgrasses (Poaceae: Triticeae) contribute substantially to aphid survival, crested wheatgrass (Agropyron spp.) particularly. In greenhouse studies, the number of aphids per plant was measured after four infestation periods on unvernalized and vernalized wheatgrasses. Before placement on these test plant species, aphids were reared either on winter wheat or on the grass host species on which aphid progeny were counted. On vernalized plants, aphids reared on wheat resulted in more aphids per test plant than when the aphids were reared on wheatgrasses, but on unvernalized plants the number of aphids per test plant did not differ significantly regardless of rearing host. Aphids on crested wheatgrass were similar in number to the other grasses when plants were unvernalized. However, when plants were vernalized, crested wheatgrass supported significantly more aphids than some of the other hosts. Aphid numbers increased on all test species as infestation period lengthened, and plant growth was largely unaffected by aphid feeding. These results suggest if sufficient moisture is available during summer when small grains are not in cultivation, all host species observed are capable of sustaining aphids. Crested wheatgrass is an abundant and important host of the Russian wheat aphid in its northern range of the western United States, but other less prevalent wheatgrasses also may contribute to aphid survival during late summer when small grains are not in cultivation. PMID- 10826181 TI - Antifeeding activity of isoquinoline alkaloids identified in Coptis japonica roots against Hyphantria cunea (Lepidoptera: Arctiidae) and Agelastica coerulea (Coleoptera: Galerucinae). AB - The anti-feeding activity of 3 isoquinoline alkaloids identified from roots of Coptis japonica Makino toward 4th-instar larvae of Hyphantria cunea Drury and adults of Agelastica coerulea Baly was examined using the leaf-dipping bioassay. The biologically active constituents of the Coptis roots were characterized as the isoquinoline alkaloids berberine, palmatine and coptisine by spectroscopic analysis. In a test with H. cunea larvae, the anti-feeding activity was much more pronounced in an application of a mixture of palmatine iodide and berberine chloride (1:1, wt:wt) at 250 ppm (82.3%) and 500 ppm (100%), compared with palmatine iodide (76.0%) and berberine chloride (75.4%) alone at 500 ppm. These results indicate a synergistic effect. With A. courulea adults, berberine chloride showed 57.5 and 91.1% anti-feeding activity at 125 and 250 ppm, respectively; whereas, weak activity was obtained in application of 500 ppm of palmatine iodide (41.4%) and coptisine chloride (52.4%) alone. The Coptis root derived compounds merit further study as potential insect-control agents. PMID- 10826182 TI - Effects of intensive forest management practices on insect infestation levels and loblolly pine growth. AB - Intensive forest management practices have been shown to increase tree growth and shorten rotation time. However, they may also lead to an increased need for insect pest management because of higher infestation levels and lower action thresholds. To investigate the relationship between intensive management practices and insect infestation, maximum growth potential studies of loblolly pine, Pinus taeda L., were conducted over 4 yr using a hierarchy of cultural treatments. The treatments were herbaceous weed control (H), H + irrigation (I), H + I + fertilizer (F), and H + I + F + pest control (P). These treatments were monitored for differences in growth and insect infestation levels related to the increasing management intensities. The Nantucket pine tip moth, Rhyacionia frustrana (Comstock), was consistently found infesting study trees. In the third field season, the H + I + F + P treatment had significantly more southern pine coneworm, Dioryctria amatella (Hulst), attacks than the H and H + I treatments. There were significant differences in volume index (D2H) among all treatments after each of the four growing seasons. This study indicated that tree fertilization can increase coneworm infestation and demonstrated that tip moth management can improve tree growth initially. Future measurements will determine if the growth gains from tip moth management are transitory or sustainable. PMID- 10826183 TI - Tomicus piniperda (Coleoptera: Scolytidae) emergence in relation to burial depth of brood logs. AB - The pine shoot beetle, Tomicus piniperda (L.), is an exotic pest of pines, Pinus spp., that was first found in the United States in 1992. A federal quarantine currently regulates movement of pine Christmas trees and pine nursery stock from infested to uninfested counties. The current national Pine Shoot Beetle Compliance Management Program requires T. piniperda-infested brood material to be disposed of by burning, chipping, or burial. The burial option requires that the infested pine material be buried at a depth of at least 30 cm. We tested this requirement by burying logs with similar levels of infestation at 0, 15, 30, 45, 61 and 76 cm and then monitoring for T. piniperda emergence. Logs were buried at two times during larval development (early and late) and in two soil types (sandy loam and loam). Emergence patterns from the two soil types were similar. Overall, 1,747 T. piniperda adults were collected from the 24 exposed control logs, but only 34 adults from the 120 buried logs, including 24 adults from logs buried at 15 cm, eight adults from 30 cm, one adult from 45 cm, and one adult from 61 cm. In comparing mean emergence density from buried logs with that of exposed logs, 98.6% mortality occurred at 15 cm, 99.5% at 30 cm, and > 99.9% at > or = 45 cm. Mean date of T. piniperda emergence to the soil surface was affected by burial depth and burial date, but not soil type. PMID- 10826184 TI - Attraction of adult Rhagoletis indifferens (Diptera: Tephritidae) to unbaited and odor-baited red spheres and yellow rectangles. AB - Five sizes of red spheres (4, 6, 8, 10, and 12 cm diameter) and 2 orientations of yellow rectangles (vertical and V) were evaluated as unbaited sticky-coated traps for western cherry fruit flies, Rhagoletis indifferens Curran, in unmanaged cherry trees in Washington and Oregon. Red spheres that were 10 cm in diameter attracted more flies than red spheres that were 8 or 12 cm in diameter and significantly more flies than 4- or 6-cm spheres and yellow rectangles of either orientation. In a 2nd test, red spheres (10 cm diameter) baited with ammonium carbonate alone or ammonium carbonate plus putrescine attracted significantly more R. indifferens than similar spheres baited with ammonium acetate alone, putrescine alone, 3-methyl-1-butanol alone, or combinations of these substances. In a 3rd test, vertical yellow rectangles baited with ammonium carbonate alone attracted numerically more R. indifferens than any of the aforementioned substances alone or in combination. We discuss the potential value of 10-cm red spheres baited with ammonium carbonate for monitoring and direct control of R. indifferens. PMID- 10826185 TI - Whole-plant CO2 exchange measurements on azaleas injured by azalea lace bug (Heteroptea: Tingidae) feeding. AB - Whole-plant gas exchange was measured continuously for 24 h on rooted cuttings of Girard's 'Pleasant White' azaleas. Azalea treatments were azalea lace bug, Stephanitis pyrioides (Scott), feeding injury levels that averaged 6, 13, or 31% leaf-area injury throughout the plant canopies. Gas exchange parameters, including net photosynthesis, dark respiration, carbon use efficiency, and growth, were compared with undamaged control plants. Responses of Girard's 'Pleasant White' azaleas suggested that azaleas were tolerant of lace bug feeding injury levels above the aesthetic threshold. Azalea tolerance can be incorporated into an integrated management plan to reduce chemical inputs into the urban landscape. PMID- 10826186 TI - Field evaluation of integrated pest management-compatible pesticides for the citrus leafminer Phyllocnistis citrella (Lepidoptera: Gracillariidae) and its parasitoid Ageniaspis citricola (Hymenoptera: Encyrtidae). AB - Potentially selective and integrated pest management (IPM)-compatible pesticides for the citrus leafminer Phyllocnistis citrella Stainton and its parasitoid Ageniaspis citricola Logvinovskaya were compared under nursery field conditions at Gainesville, FL. In 1996, replicated blocks of young grapefruit trees were treated with 2% petroleum oil and 1 x the lowest recommended field rate (LRFR) of diflubenzuron + oil (0.4%). Untreated and treated controls (avermectin at 1 x the LRFR + oil) were included. In 1997, blocks were treated with oil (3%), 1 x the LRFR of azadirachtin + oil, 1 x the LRFR of diflubenzuron + oil and 0.1 x the LRFR of avermectin + oil. Untreated and treated controls were again included. Oil at 3%, azadirachtin at 1 x the LRFR + 0.4% of oil, and diflubenzuron at 1 x the LRFR + 0.4% of oil were shown to be IPM-compatible pesticides. In 1997, these blocks had fewer mines per leaf and P. citrella pupae parasitized by A. citricola per total leaves sampled compared with the untreated control but more than the treated control (alpha = 0.05). Avermectin at 0.1 x the LRFR + 0.4% of oil was not considered an IPM-compatible pesticide because, while it reduced the number of P. citrella mines per leaf, it reduced the number of A. citricola to levels as low as the treated control. Actual P. citrella infestation levels had no detectable effect on tree growth and vigor. Pesticide applications were not justified when P. citrella infestations were < 1 mine per leaf and the biological control agent A. citricola was present. PMID- 10826187 TI - Lawn parameters influencing abundance and distribution of the hairy chinch bug (Hemiptera: Lygaeidae). AB - Management of lawns that promotes conditions detrimental to the development of insect pests may represent a valuable environmentally benign turfgrass management strategy. In the cool-humid region of Quebec, Canada, we investigated 45 lawns infested with hairy chinch bug, Blissus leucopterus hirtus Montandon, to identify lawn parameters related to its distribution and abundance. Kentucky bluegrass, creeping bentgrass, and perennial ryegrass, respectively, accounted for 55.8, 19.6, and 9.3% of the grass species. Chinch bug population density was associated positively with abundance of perennial ryegrass, whereas it was marginally negatively related with the abundance of creeping bentgrass. An index of the severity of chinch bug infestation was obtained for each lawn by combining estimates of number of infested patches per lawn, average size of the patches, and chinch bug number per patch. The index was associated positively with abundance of Kentucky bluegrass and perennial ryegrass. There was evidence that abundance of creeping bentgrass was associated negatively with the number of infested patches per lawn, area of the patches, and number of chinch bugs within those patches. The number of infested patches increased, whereas patch area and chinch bug number per patch tended to decrease, when broad-leaf weeds were more abundant on a lawn. No significant relationship was found between thatch thickness and patterns of chinch bug abundance and distribution. These results suggest that management of lawns to respectively increase and decrease abundance of creeping bentgrass and perennial ryegrass could facilitate control of hairy chinch bug populations in cool-humid regions. PMID- 10826188 TI - Citrus leafminer (Lepidoptera: Gracillariidae) density: effect on yield of 'Tahiti' lime. AB - The relationship between damage by citrus leafminer, Phyllocnistis citrella Stainton, and 'Tahiti' lime yield were investigated in a 15-yr-old and a 5-yr-old lime orchard. Citrus leafminer population densities were controlled by insecticide applications of abamectin plus FC 435 oil, abamectin plus FC435 oil plus imidacloprid, and methomyl. The control was not treated. To ensure adequate citrus leafminer densities, adult citrus leafminer were periodically released in the experimental plots during fall and winter. For the 15-yr-old trees, the least amount of leaf area damage occurred in the abamectin plus FC 435 oil plus imidacloprid (1.9%) and the abamectin plus FC435 oil (2.3%) treatments compared with the control treatment (10-21%). In the 5-yr-old orchard, the least amount of leaf area damaged occurred in the abamectin plus FC 435 oil plus imidacloprid (0.4%) and the imidacloprid (0.1%) treatments compared with the control (20.85%). The percentage of leaf area damaged was linearly correlated with the average number of mines per leaf, average mine days, and cumulative mine days in both orchards. In both orchards, the percentage of leaf area damaged and cumulative mine days was linearly correlated with the number of fruit per tree and total fruit weight per tree. Calculating the economic injury levels indicated that 16 23% and 18-85% of leaf area damaged caused significant yield reductions in 15-yr old and 5-yr-old trees, respectively. PMID- 10826189 TI - Larval growth of Diaprepes abbreviatus (Coleoptera: Curculionidae) and resulting root injury to three citrus varieties in two soil types. AB - Larval growth and intraspecific competition of Diaprepes abbreviatus (L.) larvae and consequent root injury in container-grown citrus in the greenhouse were evaluated. Roots of Carrizo citrange, Citrus sinesis L. Osbeck x Poncirus trifoliata (L.) Raf.; Cleopatra mandarin, C. reticulata Blanco, and Swingle citrumelo, C. paradisi Macf. x P. trifoliata (L.) Raf. rootstock seedlings grown in Candler fine sand and potting soil were colonized with different populations of D. abbreviatus larvae. Larvae were exposed to the seedlings for 79 d. Larval growth and development increased steadily for approximately 70 d on all rootstock soil combinations, at which time most larvae were instars 6-8. Most feeding injury occurred to roots when larvae were between instars 3 and 6. Larval weight reached a plateau at approximately 70 d, but often declined between 70 and 79 d. When larvae were small, injury to seedlings developed slowly, primarily on fibrous roots, then feeding increased rapidly, often resulting in total consumption of both fibrous root and bark tissue. Although not statistically significant, root injury developed slightly slower on Swingle citrumelo compared with Carrizo and Cleopatra rootstocks, but damage was comparable by 79 d. Little or no difference in consumptive benefit to the larvae was found between the rootstocks. Based on larval weight days, little feeding injury occurred during the first 21 d, but increased rapidly between 21 and 60 d. Soil type affected the rate of larval growth and development, with potting soil contributing to greater growth rates. Detritus in potting soil provided little or no nutritional resource, suggesting that the effect of potting soil on larval development was primarily physical. In addition, fewer inoculated larvae per seedling exhibited greater weight gains than higher infestation densities, suggesting that intraspecific competition for nutritional resources influenced larval development. PMID- 10826190 TI - Sublethal effects of resin glycosides from the periderm of sweetpotato storage roots on Plutella xylostella (Lepidoptera: Plutellidae). AB - Resin glycoside material extracted from the periderm tissue of storage roots from sweetpotato, Ipomoea batatas (L.) Lam., was bioassayed for effects on survival, development, and fecundity of the diamondback moth, Plutella xylostella (L.). The resin glycoside was incorporated into an artificial diet and fed to P. xylostella larvae. First instars were placed individually into snap-top centrifuge vials containing artificial diet with one of six concentrations of resin glycoside material (0.00, 0.25, 0.50, 1.00, 1.50, and 2.00 mg/ml). Each replication consisted of 10 individuals per concentration, and the experiment was repeated 13 times. Vials were incubated at 25 degrees C and a photoperiod of 14:10 (L:D) h in a growth chamber. After 6 d, surviving larvae were weighted and their sex determined, then returned to their vials. Later, surviving pupae were weighed and incubated at 25 degrees C until moths emerged. Females were fed, mated with males from the laboratory colony, and allowed to lay eggs on aluminum foil strips. Lifetime fecundity (eggs/female) was measured. There were highly significant negative correlations between resin glycoside levels and survival, and between glycoside levels and larval weight after 6 d of feeding. For larvae that lived at least 6 d, there was no additional mortality that could be attributed to the resin glycoside material. However, there was a significant positive correlation between glycoside dosages and developmental time of larvae (measured as days until pupation). Lifetime fecundity also was negatively affected at sublethal doses. Resin glycosides may contribute to the resistance in sweetpotato breeding lines to soil insect pests. PMID- 10826191 TI - Genetic sexing strains of mediterranean fruit fly (Diptera: Tephritidae): quality in mass-reared temperature-sensitive lethal strains treated at high temperatures. AB - The high-temperature treatment of eggs of mass-reared tsl genetic sexing strains in Mediterranean fruit fly, Ceratitis capitata (Wiedemann), during late embryogenesis (the low-high protocol) conserves more male flies than treatment during early embryogenesis. A tsl strain, AUSTRIA 6-97, was constructed to follow the fate of aneuploid individuals during male-only production. Aneuploid individuals are produced following segregation in the translocation heterozygous males, and they can survive to the pupal stage where they compromise quality because they do not eclose as adults. Hatching, emergence, and male fly production were quantified and the heat-treatment protocol was characterized. The low-high egg treatment conserves the number of euploid-balanced males, and there is a very low survival of aneuploid males. After heat treatment of eggs, at least 95% of the male pupae were euploid compared with only 71% from untreated eggs. The quality of euploid male pupae was diminished with successive daily collections, an effect previously attributed to aneuploid survivors. Reduced yield of euploid males from early heat treatments was the result of an emergence effect, in addition to a maternal effect. A third detrimental effect of heat was found, occurring after hatching and before pupation, that reduces the survivorship of euploid males. The low-high treatment protocol yielded more males, with a higher accuracy than other heat treatments. However, although it avoids both the maternal and emergence effects, the production of euploid males was 30% less than the potential production, implying that the low-high heat protocol for killing female embryos in tsl genetic sexing strains can be fine tuned. PMID- 10826192 TI - Toxicity and residual effectiveness of insecticides on insecticide-treated spheres for controlling females of Rhagoletis pomonella (Diptera: Tephritidae). AB - This study evaluated the toxicity of five technical-grade insecticides of four different classes to apple maggot females, Rhagoletis pomonella (Walsh), following a 10-min exposure period in insecticide-coated glass jars, with or without a feeding stimulant (sucrose) present. According to LC90 values for toxicity by ingestion and tarsal contact, imidacloprid was 1.5 times more toxic than dimethoate or abamectin, diazinon was less toxic, and phloxine B (a phototoxic dye) least toxic. Based on LC90 values for tarsal contact alone, dimethoate was 2.3, 4.0, and 18.4 times more toxic than imidacloprid, abamectin, and diazinon, respectively. Contact alone with phloxine B caused no mortality. When exposure was assessed using spheres coated with a latex paint mixture containing sucrose and formulated dimethoate (Digon 400 EC) or imidacloprid (Provado 1.6 F) at concentrations ranging from 5 to 70 g (AI)/cm2, both insecticides showed reduced effectiveness compared with toxicities from glass jar tests, with Digon two times more toxic than Provado. After exposure to artificial rainfall and retreatment with sucrose, Digon- and Provado-treated spheres exhibited greatest residual effectiveness, with diazinon-treated spheres less effective. Spheres treated with formulated abamectin (Agri-Mek 0.15 EC) at 1.0% (AI) performed only slightly better than phloxine B-treated spheres, which completely lost effectiveness after exposure to rainfall. Spheres treated with formulated imidacloprid (Merit 75 WP) at 1.5% (AI) showed equal or better residual efficacy in killing apple maggot flies (> 80% mortality, shorter lethal duration of feeding) over a 12-wk exposure period to outdoor weather than spheres treated with Digon at 1.0% (AI) after both types were retreated with sucrose. Our results indicate that imidacloprid is a promising safe substitute for dimethoate as a fly killing agent on lure-kill spheres. Imidacloprid formulated as Merit 75 WP had greater residual efficacy than imidacloprid formulated as Provado 1.6 F. PMID- 10826193 TI - Measuring wood consumption by subterranean termites (Isoptera: Rhinotermitidae) with digitized images. AB - A method is described in which consumption by termites of a feeding block composed of 36 thin (2-mm) wooden slats is estimated with a video image analysis. The significant regression between dry wood weight loss and decreased pixel count of digitized images demonstrates that the video image technique may be a useful tool to quantify termite feeding. Time required to measure wood consumption with the video image technique (approximately 11.5 min per sample) was significantly less than that for the manual method of washing, and weighing wooden blocks damaged by termite feeding (approximately 21.0 min per sample). PMID- 10826194 TI - Control of Coptotermes havilandi (Isoptera: Rhinotermitidae) with hexaflumuron baits and a sensor incorporated into a monitoring and baiting program. AB - A sensor consisting of a wooden monitor painted with a conductive circuit of silver particle emulsion was placed in a monitoring station to detect feeding activity of the subterranean termite Coptotermes havilandi Holmgren. Sensor accuracy was 100% 1 mo after installation, but 9 mo after sensor placement, the rate declined to 73%. After the detection of C. havilandi in the stations, baits containing the chitin synthesis inhibitor hexaflumuron were applied in five colonies, and four colonies were eliminated within 3-5 mo. Baiting could not be completed for the remaining one colony because the site became inaccessible. PMID- 10826195 TI - Feeding inhibition and mortality in Reticulitermes flavipes (Isoptera: Rhinotermitidae) after exposure to imidacloprid-treated soils. AB - Feeding inhibition and mortality of Reticulitermes flavipes (Kollar) exposed to sand, sandy loam, loam, and silty clay loam soils treated with several concentrations of imidacloprid were studied using bioassay techniques under laboratory conditions. Termite workers stopped feeding after exposure to treated soils. Differences in feeding reduction varied among the soil types. Based on the magnitude of the F-statistics, the effect of imidacloprid on the reduction of termite feeding was greatest in sand followed by sandy loam, loam, and silty clay loam soils. Soil properties such as organic matter content, silt and clay proportions, pH, and cation exchange capacity were suggested to affect the bioavailability of imidacloprid. Similar soil effects on mortality were observed in termites continuously exposed to treated soil for 21 d. In three of four soils tested, susceptibility to imidacloprid was not affected by the source of the termites tested. PMID- 10826196 TI - Evaluation of hexaflumuron for controlling the subterranean termite Coptotermes curvignathus (Isoptera: Rhinotermitidae) in Malaysia. AB - Cellulosic bait matrices containing 0.5% hexaflumuron were tested against field colonies of the subterranean termite Coptotermes curvignathus Holmgren in Malaysia. Foraging activities of the termites were first monitored for several months by using survey stakes and wood bundles placed in underground monitoring traps. Infested stakes were replaced with bait devices. Termite workers to act as recruits were collected from infested stakes and placed in the bait devices. Subsequent activities of the termites were monitored by examining and weighing the wood in the monitoring traps. Data show that the four colonies had populations of 166,288-709,052 foragers and mean wood consumption rates of 305.9 508.2 g/mo per trap. These colonies ceased their activities at all monitoring devices 25-44 d after baiting had commenced. Colonies consumed approximately 27 79 g of bait matrix or 137.5-395 mg of hexaflumuron. Thus, C. curvignathus colonies could be monitored, characterized, baited, and foraging eliminated by using a bait matrix containing hexaflumuron. PMID- 10826197 TI - Secondary transmission of toxic baits in German cockroach (Dictyoptera: Blattellidae). AB - The death rates of three toxic gel baits (fipronil bait, 0.05%; hydramethylnon, bait 2.15%; and abamectin B1 bait, 0.05%) were estimated in Blattella germanica (L.). After ingestion, all three baits killed similar proportions of cockroaches, but they died more rapidly after ingestion of fipronil bait than after ingestion of abamectin B1 bait or of hydramethylnon bait. Laboratory experiments evaluated the impact of secondary transmission of fipronil bait and hydramethylnon bait. Mortality rates caused by secondary transmission were higher for fipronil bait than for hydramethylnon bait. Under controlled laboratory conditions, secondary transmission occurred mainly through direct contact with, or ingestion of, traces of baits dispersed in the environment by contaminated cockroaches. These traces were either deposited by "trampling" in the environment or on dead contaminated cockroaches. Social interactions and cannibalism played a minor role in secondary transmission of these two baits. PMID- 10826198 TI - Seasonal changes in pyrethroid resistance in tarnished plant bug (Heteroptera: Miridae) populations during a three-year period in the delta area of Arkansas, Louisiana, and Mississippi. AB - Tarnished plant bugs, Lygus lineolaris (Palisot de Beauvois), were collected from weeds at 71 locations in the delta of Arkansas, Louisiana, and Mississippi and tested with a discriminating dose bioassay for pyrethroid resistance in the spring (April-May) and again at the same locations in the fall (September October) in 1995-1997. Percentage of mortality in the discriminating dose bioassay declined significantly (pyrethroid-resistance increased) from spring to fall by an average 18.7, 21.3, and 21.7% in Arkansas, Louisiana, and Mississippi, respectively. Pyrethroid resistance declined significantly by 26.3% from the fall of 1995 to the spring of 1996 in Mississippi, but did not significantly decline in Arkansas (4.1%) and Louisiana (13.2%). Significant decreases in resistance occurred in all 3 states from the fall of 1996 to the spring of 1997 (17.1, 38.3, and 29.8% in Arkansas, Louisiana, and Mississippi, respectively). Plant bugs from 2 locations (Indianola, MS, and Wainwright, LA) had multiple insecticide resistance to a carbamate, 2 pyrethroid, and 4 organophosphorus insecticides. However, resistance to the organophosphate acephate in plant bugs from both locations was not significant. Possible causes for the significant increases and declines in resistance from season to season are discussed. PMID- 10826199 TI - Baseline susceptibility to imidacloprid and cross resistance patterns in Colorado potato beetle (Coleoptera: Chrysomelidae) populations. AB - During 1995-1998, we tested 134 geographically discrete populations of Colorado potato beetle, Leptinotarsa decemlineata (Say), from the United States, Canada, Germany, France, and Poland for susceptibility to imidacloprid. Neonates were assayed on potato-based agar diet incorporated with imidacloprid and exposed on filter paper to esfenvalerate, azinphosmethyl, and carbofuran to characterize cross-resistance. In all 4 yr, Long Island populations were the most tolerant to imidacloprid, with LC50s ranging up to 29 times higher than the most susceptible populations. Responses to imidacloprid did not change significantly on farms where populations were assayed over time, except for those from Long Island, which doubled in overall tolerance to imidacloprid since 1995. Much of this tolerance was already present before imidacloprid was used on Long Island. Correlative analysis of the populations tested over the 4 yr indicated positive cross-resistance patterns with esfenvalerate and azinphosmethyl. This response was probably caused by preexisting metabolic and excretion mechanisms selected by previous exposure. There was no significant pattern of cross-resistance with carbofuran or bensultap. Regression slopes were also significantly negatively correlated with LC50 values for imidacloprid, indicating higher heterogeneity, which could lead in further resistance development. We discuss the relative sensitivity of diet-incorporated assays with neonates compared with other bioassay studies. Based on a pooled group of susceptible populations tested in 1995, a baseline LC50 of 0.39 ppm and a discriminating concentration of 8 ppm were suggested to detect early stages of resistance in "suspect" populations. We also suggest application strategies for imidacloprid that reduce selection pressure. PMID- 10826200 TI - Susceptibility of Spanish populations of the corn borers Sesamia nonagrioides (Lepidoptera: Noctuidae) and Ostrinia nubilalis (Lepidoptera: Crambidae) to a Bacillus thuringiensis endotoxin. AB - Baseline susceptibility to the Cry1Ab delta-endotoxin from Bacillus thuringiensis (Berliner) was determined for four populations of Sesamia nonagrioides (Lefebvre) and two populations of Ostrinia nubilalis (Hubner) from Spain. This study shows that S. nonagrioides is at least as susceptible as O. nubilalis to B. thuringiensis Cry1Ab protein. We found small differences in susceptibility among the Spanish populations of S. nonagrioides that can be attributed to natural variation, because there are no records of B. thuringiensis products being used on corn crops in Spain. There were no differences in susceptibility to Cry1Ab toxin between the two populations of O. nubilalis. PMID- 10826201 TI - Mechanisms associated with methiocarb resistance in Frankliniella occidentalis (Thysanoptera: Thripidae). AB - Biochemical mechanisms associated with methiocarb resistance were examined in laboratory-selected and field populations of the western flower thrips, Frankliniella occidentalis (Pergande). Seven populations were examined and they differed in their susceptibility to methiocarb by 30 times. Including the synergists piperonyl butoxide, a cytochrome P-450 monooxygenase inhibitor, or S,S,S-tributylphosphorotrithioate, an esterase inhibitor, in the methiocarb bioassays partially suppressed resistance in the most resistant populations. In vitro assays of general esterase, glutathione S-transferase, and acetylcholinesterase activities showed increased activity in some of the resistant populations and increased activity of the enzymes after methiocarb selection on one of the populations. Assays of acetylcholinesterase sensitivity to inhibition by methiocarb, dichlorvos, and eserine suggested insensitive acetylcholinesterase in two of the resistant populations. These results indicate that methiocarb resistance in F. occidentalis was polyfactorial and involved detoxification and altered target site. None of the biochemical assays showed interpopulation enzymatic differences strongly correlated with the level of methiocarb resistance. The possibilities for developing rapid biochemical diagnostic assays to detect methiocarb resistance in F. occidentalis are discussed. PMID- 10826202 TI - Evaluation of potato tuber moth (Lepidoptera: Gelechiidae) resistance in tubers of Bt-cry5 transgenic potato lines. AB - The potato tuber moth, Phthorimaea operculella (Zeller), in tropical and subtropical countries, is the most destructive pest of potato, Solanum tuberosum L. The larvae attack foliage and tubers in the field and in storage. The purpose of this study was to evaluate the efficacy of a Bt-cry5 transgene to control the potato tuber moth in tuber tissues. Tuber bioassays using stored (11-12 mo old) and newly harvested tubers of Bt-cry5-Lemhi Russet and Bt-cry5-Atlantic potato lines showed up to 100% mortality of 1st instars. Mortality was lowest in the newly harvested tubers of Bt-cry5-Atlantic lines (47.1-67.6%). Potato tuber moth mortality was 100% in the Bt-cry5-Spunta lines that were transformed with Bt-cry5 gene controlled by the CaMV 35S promoter (pBIML5 vector) and in 2 of 3 lines transformed with Bt-cry5 gene controlled by the Gelvin super promoter (pBIML1 vector). The transgenic Spunta lines expressing Bt-cry5 controlled by the patatin promoter (pBMIL2 vector) showed the lowest tuber moth mortality (25.6 and 31.1%). The Bt-cry5 transgenic lines with high tuber expression of B. thuringiensis have value in an integrated pest management system to control potato tuber moth. PMID- 10826203 TI - Influence of whorl region from resistant and susceptible corn genotypes on fall armyworm (Lepidoptera: Noctuidae) growth and development. AB - The effect of diets prepared from whorl tissue of resistant and susceptible corn genotypes, Zea mays L., on the larval growth, development, and physiology of fall armyworm, Spodoptera frugiperda (J. E. Smith), was analyzed. Larvae reared on an optimized artificial diet had a higher growth rate and developed faster than those reared on lyophilized whorl tissue from resistant and susceptible genotypes. Larvae reared on the resistant material were smaller and had a longer developmental period. Larvae reared on yellow-green and green whorl sections from resistant plants were significantly smaller than those reared on the same sections of susceptible plants. There was no significant difference in weight when larvae were reared on the yellow whorl regions from either resistant or susceptible lines. Physiological indices were determined for larvae fed resistant and susceptible lyophilized and fresh whorl material. Larvae fed resistant lyophilized material had significantly lower growth rate (GW) and efficiency of conversion of ingested food to body substance (ECI) than those reared on artificial diet or susceptible material. However, there were no significant differences in consumption index (CI), approximate digestibility (AD) and efficiency of conversion of digested food to body substance (ECD) between larvae reared on lyophilized tissue from resistant and susceptible genotypes. Larvae reared on fresh yellow-green whorl sections from resistant plants had significantly lower GW, ECI, and ECD than those reared on susceptible material. In contrast, no significant differences in any of the estimated food consumption and utilization indices were observed between larvae reared on fresh yellow whorl sections from resistant or susceptible plants. These results suggest that some components of whorls from resistant plants, especially the yellow-green region, inhibit food utilization in fall armyworm larvae. PMID- 10826204 TI - Effect of plant age, larval age, and fertilizer treatment on resistance of a cry1Ab-transformed aromatic rice to lepidopterous stem borers and foliage feeders. AB - The resistance of vegetative, booting, and flowering stage plants of a variety of an aromatic rice, Oryza sativa L., transformed with a Bacillus thuringiensis Berliner cry1Ab gene under control of the maize phosphoenolpyruvate carboxylase (PEPC) promoter was evaluated against four lepidopterous rice pests--the stem borers Chilo suppressalis (Walker) (Lepidoptera: Crambidae) and Scirpophaga incertulas (Walker) (Lepidoptera: Pyralidae), and the foliage feeders Cnaphalocrocis medinalis Guenee (Lepidoptera: Pyralidae) and Naranga aenescens Moore (Lepidoptera: Noctuidae). Plants of the cry1Ab-transformed line (no. 827) were more resistant to young larvae of S. incertulas, C. suppressalis, and C. medinalis than control plants at the vegetative stage but not at the flowering stage. Survival of 10-d-old stem borer larvae did not differ on cry1Ab plants and control plants at either the vegetative or flowering stage, but the development of 10-d-old C. suppressalis larvae was retarded on the vegetative stage cry1Ab plants. Immunological analysis also showed an apparent decline in Cry1Ab titer in leaf blades and leaf sheaths at the reproductive stage. In experiments comparing three fertilizer treatments (NPK, PK, and none), there was a significant interaction between fertilizer treatment and variety on larval survival only in whole-plant assays at booting stage with C. suppressalis. On cry1Ab plants, larval survival did not differ significantly among the three fertilizer levels, whereas on control plants survival was highest with the NPK treatment. cry1Ab plants tested at the sixth and seventh generations after transformation were more resistant than control plants to N. aenescens and C. suppressalis, respectively, suggesting that gene silencing will not occur in line 827. The results of the experiments are discussed in terms of resistance management for B. thuringiensis toxins in rice. PMID- 10826205 TI - Evaluation of sampling methodology for determining the phenology, relative density, and dispersion of western flower thrips (Thysanoptera: Thripidae) in nectarine orchards. AB - Western flower thrips, Frankliniella occidentalis (Pergande), cause serious economic damage to nectarines in the Okanagan and Similkameen Valleys, British Columbia, Canada. We evaluated several sampling methods for western flower thrips for their precision and ability to predict general population trends. Beating of branches, flicking of buds, and visual estimation methods were not accurate for estimating numbers of thrips in nectarine buds. Thrips caught on sticky cards indicated general population trends, but were less efficient than collecting nectarine buds and counting thrips. Searching for thrips from buds in the field underestimated the density of both adults and larvae, and for adults, underestimated the proportion of the pale morph of western flower thrips. Dispersion patterns of thrips populations among orchards were either random or aggregated dependent on the development stage of the nectarine buds. PMID- 10826206 TI - Spatial distribution and sampling plans for Thrips palmi (Thysanoptera: Thripidae) infesting fall potato in Korea. AB - The spatial distribution of adult and immature Thrips palmi Karny on fall potato, Solanum tuberosum L., on Cheju Island, Korea, was studied over a 2-yr period by visually inspecting potato leaves. The majority of thrips collected from the leaves were observed in the top one-third of the plant. The within-field spatial patterns of adults and immature thrips were aggregated. The slopes and intercepts of Taylor's power law did not differ among adults and immature thrips. A fixed precision-level sampling plan was developed using the parameters from Taylor's power law and was tested with resampling simulations using eight independent data sets. Over a wide range of densities, the simulation demonstrated that actual sampling precision (d = SEM/mean) values at d = 0.25 averaged < 0.24 in all cases. A binomial sampling plan for estimating mean density was developed using an empirical model evaluated at tally thresholds (the minimum number of insects present before a leaf is considered infested) of one, three, five, and eight thrips per leaf. Increasing sampling size had little effect on the precision of the estimated mean regardless of tally threshold (T). However, increasing T had a dramatic effect on precision. The best tally threshold for estimating thrips density based on the applicable density ranges and the precision of the model was T = 5. A binomial sampling plan with a tally threshold of five and a fixed sample size of 30 leaves should be an effective replacement for enumerative counts when thrips average < 10 per leaf. PMID- 10826207 TI - Statistical inference on associated fertility life table parameters using jackknife technique: computational aspects. AB - Knowledge of population growth potential is crucial for studying population dynamics and for establishing management tactics for pest control. Estimation of population growth can be achieved with fertility life tables because they synthesize data on reproduction and mortality of a population. The five main parameters associated with a fertility life table are as follows: (1) the net reproductive rate (Ro), (2) the intrinsic rate of increase (rm), 3) the mean generation time (T), (4) the doubling time (Dt), and (5) the finite rate of increase (lambda). Jackknife and bootstrap techniques are used to calculate the variance of the rm estimate, which can be extended to the other parameters of life tables. Those methods are computer-intensive, their application requires the development of efficient algorithms, and their implementation is based on a programming language that encompasses quickness and reliability. The objectives of this article are to discuss statistical and computational aspects related to estimation of life table parameters and to present a SAS program that uses jackknife to estimate parameters for fertility life tables. The SAS program presented here allows the calculation of confidence intervals for all estimated parameters, as well as provides one-sided and two-sided t-tests to perform pairwise or multiple comparison between groups, with their respective P values. PMID- 10826208 TI - Effect of temperature and treatment conditions on the mortality of Epiphyas postvittana (Lepidoptera: Tortricidae) exposed to ethanol. AB - Morality responses were determined for 5th-instar lightbrown apple moth, Epiphyas postvittana (Walker), in the presence and absence of apples to immersion in ethanol solutions and exposure to ethanol vapor at a range of concentrations, treatment times, and temperatures. Ethanol may have caused an initial knock-down effect in E. postvittana larvae because there was a trend for larval mortality to reduce with increased periods between treatment and assessment time when immersed at 20 degrees C in 30 or 50% ethanol solutions. Mortality for larvae immersed on apples in a range of ethanol concentrations was higher than for larvae in the absence of apples. Increasing treatment temperature from 20 to 45 degrees C during ethanol immersion significantly increased larval mortality. During ethanol vapor exposure, longer treatment times were required to achieve 99% E. postvittana mortality for larvae on apples compared with those in the absence of apples. PMID- 10826209 TI - Toxicity of diatomaceous earth to red flour beetles and confused flour beetles (Coleoptera: Tenebrionidae): effects of temperature and relative humidity. AB - Red flour beetles, Tribolium castaneum (Herbst), and confused flour beetles, Tribolium confusum (DuVal), were exposed for 8-72 h to diatomaceous earth (Protect-It) at 22, 27, and 32 degrees C and 40, 57, and 75% RH (9 combinations). Insects were exposed to the diatomaceous earth at 0.5 mg/cm2 on filter paper inside plastic petri dishes. After exposure, beetles were held for 1 wk without food at the same conditions at which they were exposed. Mortality of both species after initial exposure was lowest at 22 degrees C but increased as temperature and exposure interval increased, and within each temperature decreased as humidity increased. With 2 exceptions, all confused flour beetles were still alive after they were exposed at 22 degrees C, 57 and 75% RH. Mortality of both species after they were held for 1 wk was greater than initial mortality for nearly all exposure intervals at each temperature-humidity combination, indicating delayed toxic effects from exposure to diatomaceous earth. For both species, the relationship between mortality and exposure interval for initial and 1-wk mortality was described by linear, nonlinear, quadratic, and sigmoidal regression. Mortality of confused flour beetles was lower than mortality of red flour beetles exposed for the same time intervals for 46.7% of the total comparisons at the various temperature-relative humidity combinations. PMID- 10826210 TI - Progeny production by Tribolium castaneum (Coleoptera: Tenebrionidae) and Oryzaephilus surinamensis (Coleoptera: Silvanidae) on maize previously infested by Sitotroga cerealella (Lepidoptera: Gelechiidae). AB - A laboratory experiment was conducted to quantify the effects of infestation of maize by Sitotroga cerealella (Olivier) on progeny production by two common secondary colonizers of grain, Tribolium castaneum (Herbst) and Oryzaephilus surinamensis (L.). Adults of both secondary pest species were allowed to oviposit for 3 wk on intact kernels of 'DeKalb 689', mechanically split kernels, kernels that had been infested for 3 mo by S. cerealella, and kernels that had been infested for 6 mo. Progeny of both species reached highest numbers on 6-mo infested maize. Prior infestation for 6 mo by S. cerealella makes maize a more suitable medium for reproduction by T. castaneum and O. surinamensis, much more than can be accounted for by mere disruption of kernel integrity resulting from larval feeding. The results highlight the importance of limiting establishment by S. cerealella on maize in storage. PMID- 10826211 TI - Bioactivities of methyl allyl disulfide and diallyl trisulfide from essential oil of garlic to two species of stored-product pests, Sitophilus zeamais (Coleoptera: Curculionidae) and Tribolium castaneum (Coleoptera: Tenebrionidae). AB - Two of the major constituents of the essential oil of garlic, Allium sativum L., methyl allyl disulfide and diallyl trisulfide, were tested against Sitophilus zeamais Motschulsky and Tribolium castaneum (Herbst) for contact toxicity, fumigant toxicity, and antifeedant activity. The contact and fumigant toxicities of diallyl trisulfide were greater than that of methyl allyl disulfide to the adults of these two species of insects. These two compounds were also more toxic to T. castaneum adults than to S. zeamais adults. Older T. castaneum larvae were more susceptible to the contact toxicity of the two compound, whereas younger larvae were more susceptible to the fumigant toxicity of these compounds. Both compounds reduced egg hatching of T. castaneum and subsequent emergence of progeny. Diallyl trisulfide totally suppressed egg hatching at 0.32 mg/cm2, and larval and adult emergence at 0.08 mg/cm2. Methyl allyl disulfide significantly decreased the growth rate, food consumption, and food utilization of adults of both insect species, with feeding deterrence indices of 44% at 6.08 mg/g food for S. zeamais and 1.52 mg/g food for T. castaneum. However, it did not affect any nutritional indices of T. castaneum larvae. Diallyl trisulfide significantly reduced all of the nutritional indices in all of the insects tested. Feeding deterrence indices of 27 and 51% were obtained in S. zeamais adults and T. castaneum larvae, respectively, at the concentration of 2.98 mg/g food, whereas feeding deterrence of 85% was achieved in T. castaneum adults at a much lower concentration of 0.75 mg/g food. Hence, diallyl trisulfide is a more potent contact toxicant, fumigant and feeding deterrent than methyl allyl disulfide. PMID- 10826212 TI - Presence-absence sequential sampling plan for northern fowl mite, Ornithonyssus sylviarum (Acari: Macronyssidae), on caged-layer hens. AB - Caged-layer hens were scored as infested or uninfested by visual examination of the vent region, and the number of northern fowl mite, Ornithonyssus sylviarum (Canestrini & Fanzago), per hen was estimated. The proportion infested and average number of mites per hen were shown to have a highly significant, positive relationship (r = 0.936). Sampling among houses within a flock, and rows and sections within houses were analyzed to determine the reliability of sampling a representative portion of a flock. Low- and moderate-tolerance treatment thresholds, based on percentage of hens infested with mites, were developed from sampling 1 wk before and 1 wk after acaricide treatments determined necessary by the producer. These thresholds were used to compare a fixed (single) sampling plan, a curtailed procedure of the fixed sampling plan, and a sequential sampling plan based on a sequential probability ratio test, by sampling 174 hens (the maximum number needed for the single sampling plan). The sequential sampling plan required fewer hen examinations on average to reach a treatment decision than did the other plans, depending on the infestation tolerance limits. Using a low tolerance approach in which infestations below 15% are considered noneconomic (safe threshold) and infestations above 25% are considered economically important (action threshold), as few as 5 hens required examination to reach a treatment decision. Sequential sampling plan graphs are presented for 2 tolerance threshold scenarios (a 15% safe-threshold paired with a 25% action threshold and a 35% safe threshold paired with a 45% action threshold). These sequential sampling plans using presence absence assessments should greatly facilitate monitoring and treatment decisions for this important pest. PMID- 10826213 TI - Behavioral health expenditures and state organizational structure. AB - The authors present a study on expenditures by state mental health, substance abuse, and developmental disability agencies in the United States for the period between 1981 and 1993. The relationship between agency spending and organizational structure of state bureaucracy was examined. Results indicate that organizational structure is a determinant of agency spending. The more independent an agency, the higher its spending; conversely, the more independent its competitor, the lower the agency's spending. The number of levels between an agency and the governor's office was not significant in explaining agency expenditures. PMID- 10826214 TI - Performance improvement teams for better psychiatric rehabilitation. AB - Despite the promise of psychiatric rehabilitation, many programs fail to incorporate innovative rehabilitation practices into their day-to-day regimens. Performance improvement is an effective paradigm for helping agencies improve the quality of their programs. Four phases of performance improvement are reviewed in this article: organizing for change, preparing the environment, focusing the environment, and maintaining improvement. Implementing the four phases of performance improvement is illustrated in a case study. Methodological rigor of data generated by performance improvement teams is also discussed. PMID- 10826215 TI - Reliability of Medicaid claims files for use in psychiatric diagnoses and service delivery. AB - To investigate the reliability of Medicaid claims data for use in research, clinical decision-making, and policy, medical records were abstracted of 105 inpatient stays on the psychiatric service of a large general hospital. Primary and secondary diagnoses and outpatient specialty mental health services after hospitalization were compared between Medicaid claims data and medical record information. Primary and secondary diagnoses were reliable, but claims data failed to capture several types of outpatient services. This suggests strategies to use claims files more appropriately. PMID- 10826216 TI - Developing a case management strategy in Israel. PMID- 10826217 TI - Using panel data to model community functioning of severely mentally ill persons. PMID- 10826218 TI - Geriatric psychiatry: evolution of an inpatient unit. PMID- 10826219 TI - Legionella from guests of Welsh hotel indistinguishable from humidifier isolates. PMID- 10826220 TI - Meeting the health needs of refugees and asylum seekers. PMID- 10826221 TI - [Characteristics of auricular changes by ageing--a case report from Saga Prefecture]. AB - Measurements of auricles were done about 317 volunteers living in the Saga Prefecture at the age of 50 to 94 years old (71 men and 246 women), from July until November, 1999. 1) A growing tendency was observed in both male and female physiognomic ear length. 2) In contrast, there was no significant increase or decrease in the ear width. 3) A decreasing tendency was observed in the ear index of both male and female volunteers. PMID- 10826222 TI - [An adult case of recurrent myelopathy presenting with monoplegia following asthmatic attacks]. AB - A 22-year-old woman noticed an acute onset muscle weakness in her right leg after an asthmatic attack. A neurological examination showed moderate muscle weakness from the right iliopsoas muscle to the toe muscles, a decreased deep tendon reflex in her right lower limb and mild hypesthesia in her right L2-S1 segments. Needle EMG revealed fibrillation potentials in the right gastrocnemius muscle and a positive sharp wave in the right anterior tibialis muscle. The findings of motor nerve conduction studies and sensory nerve conduction studies were normal. The lesion was considered to be located at the posterior horns, as well as at the anterior horns at the L2-S1 levels and also at their roots on the right side. The anterior horn cells appeared to be the most severely affected. Polio-, echo-, entero- and coxsackie-virus antibody titers showed no significant changes on the 36th and 64th days of the disease. The serum IgE level was elevated and mite antigen-specific IgE was strongly positive. MRI revealed no abnormalities in either the thoracic or lumbar spinal cord. Although the sensory disturbance did rapidly improve after corticosteroid therapy, no improvement was seen in her muscle weakness which thus resulted in the atrophy of the affected muscle. While undergoing the corticosteroid therapy, she suffered another asthmatic attack. Nine days after the second attack, she further developed weakness in her right deltoid, biceps brachii, triceps brachii, wrist extensor, wrist flexor, digits extensor and digits flexor muscles with hyperreflexia in her left upper limb. Cervical MRI disclosed a high intensity area at the C3-6 level on the T2-weighted images and also a gadolinium enhancement of the lesion. Since monoplegia had appeared twice previously after bronchial asthma attacks in this case, Hopkins syndrome was suggested. Hopkins syndrome has so far been exclusively reported in children, and no recurrent cases have ever been reported with this condition. This is therefore considered to be the first case of Hopkins syndrome occurring in an adult and also demonstrating recurrence. PMID- 10826223 TI - Double blind study of the effect of caffeine on the mental calculation. AB - The double blind study of the effect of caffeine on the mental calculation has been carried out in the practical exercise course for the undergraduate students of clinical pharmacology, in the Medical School, Kyushu University. The results obtained from 1997 to 1999 were summarized and evaluated. PMID- 10826224 TI - Anti-Ri-associated paraneoplastic cerebellar degeneration without opsoclonus in a patient with a neuroendocrine carcinoma of the stomach. AB - We report a case of a 63-year-old man suffering from anti-Ri-associated paraneoplastic cerebellar degeneration (PCD) with gastric cancer. The neurologic presentation was limited to severe cerebellar ataxia without opsoclonus. The gastric cancer was composed of both poorly differentiated adenocarcinoma and neuro-endocrine carcinoma. The patient's serum reacted with recombinant Ri antigen and the neuroendocrine tumor component. It is thus considered that PCD without opsoclonus in the present case was related to the gastric neuroendocrine tumor and anti-Ri antibody. PMID- 10826225 TI - Medical response to bioterrorism: are we prepared? AB - The 1995 bombing of the Federal Building in Oklahoma City made terrorism a shocking reality for all Oklahomans. The fact that such an event could happen here, far from foreign political and ethnic conflicts and from large coastal cities, made us realize that no one is immune from the terrorist threat. But as horrific as that event and other terrorist incidents have been, the present threat of the use of infectious agents in acts of terrorism has the potential to cause far greater human death and suffering than any previously used terrorist weapon. Although health care professionals have not historically participated in terrorist planning and response activities, the addition of infectious agents to the terrorists' arsenal makes medical and public health professionals critical players in these efforts. Indeed, physicians have the ability to first recognize and report such an event and thereby diminish the impact. To meet the bioterrorism responsibility, physicians must be knowledgeable of this type of threat. In this article we present an overview of bioterrorism and review the organisms most likely to be used in a terrorist event. PMID- 10826226 TI - Screening for gestational diabetes: an evidence-based approach. AB - The primary goal of medicine should be to improve the health of patients. Evidence-based medicine is the utilization of evidence from studies published in the medical literature to help clinicians make decisions with and for their patients. Ideally the evidence should be patient-oriented, demonstrating improvements in actual patient outcomes, not just physiologic parameters or other intermediate endpoints. This concept is important in making decisions about screening tests, since this is how clinicians can determine if the screening test will actually improve patients' (and in the case of obstetrics, the neonates') outcomes. The purpose of this article is to examine screening for gestational diabetes from this perspective, discuss the current recommendations, and make suggestions for its utilization in clinical care. PMID- 10826227 TI - Smoking cessation overview. AB - This article summarizes the primary recommendations of the Agency for Health Care Quality's recently published Smoking Cessation Clinical Practice Guidelines. Specific recommendations are outlined to assist physicians in treating patients who smoke to end their use of tobacco. PMID- 10826228 TI - Reducing teen pregnancy in Oklahoma: focusing on older teens (18-19 years old). AB - Births to females aged 18 and 19 account for two-thirds of teen births in Oklahoma annually. It is important for Oklahoma to identify effective ways to reduce births to this age group. Nationally and locally, there are few prevention programs and little data. Oklahoma has the opportunity to lead the nation in reducing births to older teens by aggressively addressing the issue, sharing promising approaches, and building prevention efforts that include older teens in meaningful ways. PMID- 10826229 TI - Editorial on de-unification. PMID- 10826230 TI - Initial field testing of an instrument to measure: observable indicators of nursing home care quality. AB - The "Observable Indicators of Nursing Home Care Quality" instrument was developed as a new measure of nursing home care quality. The instrument is based on a theoretical model of quality nursing home care grounded in data from provider and consumer focus groups. The instrument was piloted in 10 Missouri nursing homes. Subsequent versions were tested in 109 Missouri and 11 Icelandic nursing homes. Content validity was established using experts. Concurrent and known groups validity was evaluated using Minimum Data Set quality indicators, survey citations, and a process of care measure. Interrater and test-retest reliabilities were calculated as well as coefficient alpha. The "Observable Indicators of Nursing Home Care Quality" instrument is a new measure that can be used by researchers, and potentially by regulators, consumers, or providers, to observe and score specific indicators of quality care following a 20- to 30 minute inspection of a nursing home. PMID- 10826231 TI - A systems approach to medication error reduction. AB - An interdisciplinary process for the reduction of medication errors was implemented at East Jefferson General Hospital in Metairie, Louisiana, a suburb of New Orleans. The Medication Administration Review and Safety Committee expanded the error reporting system with a comprehensive concurrent chart review process. Safe administration of medications through a systems approach has led to a reduction of documentation deficiencies, a new follow-up process, and an increase in reported errors. PMID- 10826232 TI - Reducing nosocomial pressure ulcers in an acute care facility. AB - In 1996, a nursing committee at an acute care facility organized the first pressure ulcer point prevalence survey for that hospital. In 1996, hospital acquired pressure ulcers were 90 percent of the predicted prevalence rate; in 1997, the rate dropped to 59 percent of the predicted prevalence and in 1998, to 53 percent of the predicted prevalence. The severity index decreased markedly from 291 (1996) to 98 (1997) then to 62 (1998). These improvements are attributed to the purposeful addition of multidimensional interventions, including best practices and research-based protocols, to prevent and treat nosocomial pressure ulcers. PMID- 10826233 TI - Evaluating the reliability and utility of cumulative intake and output. AB - During times of scarce resources available for providing patient care, it becomes necessary to validate the effectiveness of standard practices that consume caregivers' time and efforts. Three nurse managers of nursing units serving diverse patient populations studied the effectiveness of continuous intake and output in estimating daily fluid balance. They examined 48-hour records of daily weight and intake and output of 73 patients and correlated the two. Their findings suggest that even when caregiver charting compliance is optimal, daily intake and output recording provides unreliable results. The authors recommend daily weights alone be adopted for all patients who are not experiencing acute renal conditions. PMID- 10826234 TI - The experience of a community hospital in quantifying and reducing patient falls. AB - A retrospective review of patient falls in a 248-bed acute-care community hospital was conducted in order to quantify the number of patient falls and identify what factors resulted in these falls. The author reports these results and describes specific measures that were implemented in an attempt to reduce the number of falls in the organization. PMID- 10826235 TI - Establishment of interrater reliability for a nursing Extrapyramidal Side Effects (EPS) Assessment Scale. AB - Many patients who receive antipsychotic medications experience dystonia, akinesia, dyskinesia, and akathisia, collectively called Extrapyramidal Symptom Side Effects (EPS). The purpose of this study was to establish interrater reliability for a Nursing EPS Assessment Scale developed to focus on all four symptom areas. Twenty RNs and 12 patients participated in the instrument development studies. After several revisions, interrater reliability significance was demonstrated at the 0.01 level. It was concluded that the Nursing EPS Assessment Scale possessed good interrater reliability for nursing assessment of EPS. PMID- 10826236 TI - Family matters: a quality initiative through the patient's eyes. AB - Understanding the patient's experience of illness and health care requires that health care institutions understand how patients perceive their needs. Furthermore, nurses at the bed-side are at the forefront for impacting the quality of care patients receive. The department of medicine/oncology nursing of an academic medical center examined a report by the American Hospital Association/Picker Institute and integrated its findings into a departmental quality plan. In this article, the dimensions of care described by the American Hospital Association/Picker Institute will be discussed as well as a creative framework for the nurse to address patient needs. PMID- 10826237 TI - A nurse-led, unit-based program to reduce restraint use in acute care. AB - Although the effectiveness of restraints is not proven, the dangers of restraints are clear. Many advocate for the reduction of restraint use in tertiary care settings. The Joint Commission on the Accreditation of Healthcare Organizations is setting the standard by insisting that hospital administrations create a culture that emphasizes restraint reduction. Despite the importance of this task, very little is available in the literature to direct or guide this effort. The purpose of this article is to describe a unit-based Restraint Reduction Program that was successful on a neurology/neurosurgery unit. PMID- 10826238 TI - Effects of early middle ear effusion on child intelligence at three, five, and seven years of age. AB - OBJECTIVE: This study tested the hypothesis that children with prolonged middle ear effusion (MEE) during the first 3 years of life are at risk for cognitive delays or deficits. METHOD: A prospective study enrolled 698 children from diverse backgrounds and controlled for eight demographic and environmental factors. Participants were recruited at birth and monitored for ear status frequently in the home; 379 children were assessed for cognition with the Stanford-Binet, 4th ed., at 3 years of age, 294 at 5 years, and 198 at 7 years. RESULTS: Using the SAS General Linear Models (GLM) procedure, we found a significant direct relation between duration of bilateral MEE and Stanford-Binet Composite and Nonverbal Reasoning/Visualization Factor scores at age 3, but not at age 5 or age 7. Statistical clustering analysis revealed four groups with different temporal patterns of MEE: Low MEE, Early MEE (peaking at 0-6 months), Later MEE (peaking at 6-12 months), and High MEE. GLM analyses revealed no direct effects, but several moderated effects, of MEE cluster on cognitive development at 3 years, but none at 5 or 7 years. In general, children in the Later MEE and High MEE groups appeared to be more adversely affected by bilateral MEE at 3 years, but effects were moderated in complex ways by socioeconomic status or home stimulation. Growth curve modeling across the three assessment periods showed no effects of total duration of MEE but did indicate that children in the Later MEE cluster had low scores at age 3 but caught up at ages 5 and 7. CONCLUSIONS: Prolonged MEE, especially between 6 and 12 months, may put children at risk for cognitive delays at 3 years, but the risk effect is not strong and effects are no longer detectable at 5 or 7 years. PMID- 10826239 TI - Otitis media: precursor of delayed reading. AB - OBJECTIVE: To investigate the connection between otitis media in the language acquisition years and the occurrence of delayed reading between the ages of 8 and 10. METHOD: Participants were 40 children, half of whom had a history of otitis media between the ages of birth and three years and half who were free of the disease. These children, now ages 8-10, were tested with the WISC-R and a variety of reading measures. RESULTS: Children with a history of otitis media scored over a year below grade level in reading and significantly below controls on a variety of literacy measures as well as on the Verbal Comprehension factor on the WISC-R. CONCLUSIONS: Children with early onset otitis media (birth to three years) tend to be at greater risk for delayed reading than age-matched controls. PMID- 10826240 TI - Commentary: otitis media: consistency is the hobgoblin? PMID- 10826241 TI - Randomized, controlled trial of behavior therapy for families of adolescents with insulin-dependent diabetes mellitus. AB - OBJECTIVE: To describe the short-term results of a controlled trial of Behavioral Family Systems Therapy (BFST) for families of adolescents with diabetes. METHODS: We randomized 119 families of adolescents with diabetes to 3 months' treatment with either BFST, an education and support Group (ES), or current therapy (CT). Family relationships, psychological adjustment to diabetes, treatment adherence and diabetic control were assessed at baseline, after 3 months of treatment (reported here), and 6 and 12 months later. RESULTS: Compared with CT and ES, BFST yielded more improvement in parent-adolescent relations and reduced diabetes specific conflict. Effects on psychological adjustment to diabetes and diabetic control were less robust and depended on the adolescent's age and gender. There were no effects on treatment adherence. CONCLUSIONS: BFST yielded some improvement in parent-adolescent relationships; its effects on diabetes outcomes depended on the adolescent's age and gender. Factors mediating the effectiveness of BFST must be clarified. PMID- 10826242 TI - A multimethod assessment of behavioral and emotional adjustment in children with asthma. AB - OBJECTIVE: Examine behavioral adjustment and emotion regulation among 6-year-old children with asthma and a group of healthy controls. METHOD: Subjects were 81 children with asthma and 22 healthy controls. Asthma and allergy statuses were confirmed by objective measures. Emotional and behavioral functioning were assessed through parent report, child interview, and child participation in an emotional regulation paradigm. RESULTS: Maternal report revealed more internalizing and total behavior problems for children with asthma compared to controls. Child interview and behavioral observations of emotion regulation yielded no differences between groups. Severity of asthma was related to increased emotional difficulties by clinician interview and observation but not by maternal report. CONCLUSIONS: Two groups of children with asthma who have psychological difficulties include those with increased anxiety and those with poor emotion regulation and more asthma symptoms. Different measures of child adjustment yield a complex picture of the behavioral difficulties associated with pediatric asthma. PMID- 10826243 TI - Commentary: rethinking our interventions in pediatric chronic pain and treatment research. AB - The dearth of empirical support for interventions in pediatric chronic and disease-related pain requires a shift in our treatments and treatment research. Treatments with a higher degree of treatment accommodation are needed to reduce the treatment demands on patients and their families. We need to focus more on changing our treatments to fit the lives of our patients instead of trying to change our patients' lives to fit our treatments. Likewise, treatment research will need higher levels of research acceptability to ensure adequate data demonstrating the true efficacy of treatments. A more creative approach is needed that incorporates technology as a tool to these ends. PMID- 10826244 TI - [Analysis of the contractions evoked by sympathetic nerve stimulation, and thermal effect on the guinea-pig vas deferens--study as a model for the thermotherapy of benign prostatic hyperplasia]. AB - BACKGROUND: The present study was designed to investigate the mechanism of thermotherapy on benign prostatic hyperplasia (BPH), using the guinea-pig vas deferens as a model for BPH. The components of contractions elicited by electrical field stimulation and nicotine were analyzed, and the thermal effect on the vas deferens was examined. METHODS: The vas deferens was dissected, suspended vertically through two silver ring electrodes, and attached to an isometric transducer. The electrical stimulation of 10 constant current pulses (10 mA) with 0.3 msec in duration of 5, 10, and 40 Hz was achieved under air-gap condition. Drugs were added directly to a 5 ml Magnus tube containing Tyrode solution (36 degrees C) gassed with a 95% O2-5% CO2 mixture. The components of contractions evoked by electrical stimulation and nicotine were investigated by tetrodotoxin (TTX), and blocking agents of alpha 1-adrenoceptors and/or purinoceptors. Thermal effect on electrically evoked contractions was examined at incubation temperature of 25 degrees C (control), 43 degrees C, 45 degrees C, 46 degrees C and 47 degrees C for 1 hour. RESULTS: Nicotine (200 microM) elicited biphasic contractions, which were triggered by corelease of noradrenaline (NA) and ATP (N-ATP) from sympathetic nerve terminals by activation of prejunctional nicotine receptors. NA and N-ATP caused the corresponding contractions, alpha 1 and N-ATP components, respectively. Combined application of prazosin (1 microM) and suramin (50 microM) abolished these contractions. Activation of post junctional alpha 1-adrenoceptors by NA caused release of ATP from muscle cells to produce the contraction (alpha 1-ATP component), which was sensitive to both suramin and prazosin. N-ATP and alpha 1 components attributed to fast and slow part of the contraction, respectively. Electrical field stimulation caused biphasic contractions which consisted of both neurogenic (TTX-sensitive) and non neurogenic (TTX-insensitive) components. An increase in stimulation frequency (5 to 40 Hz) increased the neurogenic components, which contained alpha 1 and N-ATP components, as well as the case of nicotine. The non-neurogenic components consisted of alpha 1-ATP, muscle-derived ATP (m-ATP) and unknown substance 'X' components. Nifedipine (10 microM). L-type Ca2+ channel blocker, markedly reduced the contractions induced by bath applied phenylephrine (alpha 1-agonist, 100 microM) but only partially blocked the contractions produced by bath applied ATP (500 microM). The contractile force in amplitude and neurogenic components induced by electrical field stimulation did not change at 43 degrees C, but both declined significantly above 45 degrees C. The neurogenic components at 45 degrees C and 46 degrees C were suppressed to 22 +/- 6% and 14 +/- 3% (mean +/- SD) of control, respectively. All the contractile responses were abolished at 47 degrees C. CONCLUSION: The contractions of the guinea-pig vas deferens evoked by electrical field stimulation consisted of alpha 1, N-ATP, alpha 1-ATP, m-ATP and X components. Sympathetic nerve fibers in the muscles were completely inactivated by thermal exposure at 47 degrees C for 1 hour. The results suggest that the minimal temperature for thermotherapy of BPH should be 47 degrees C. PMID- 10826245 TI - [The problem of a long-term follow up in patients with renal cell carcinoma]. AB - PURPOSE: We investigated on a problem of long-term follow up in patients with renal cell carcinoma. PATIENTS AND METHODS: A total of 287 patients with renal cell carcinoma treated in Nara Medical University and affiliated facilities from January 1980 to December 1990 were examined. And we investigated the trend of explanation to patients including 287 patients from 1991 to 1995. RESULTS: Up to December 1995, there are 76 patients (26.5%) unable to be followed and 211 patients able to be followed. The former group patients were less declared cancer rather than the latter group patients. Of 76, 22 patients (28.9%) might misunderstand completely recovering from the cancer disease. For the recent 5 years, those patients who were declared cancer increased, and those patients who were explained benign disease decreased. CONCLUSION: These results suggested that declaration of cancer is important for patients with renal cell carcinoma to be followed for a long-term. PMID- 10826246 TI - [Congenital hydronephrosis detected by prenatal ultrasonography: the effect of pyeloplasty on renal function]. AB - PURPOSE: Prenatal detection of congenital hydronephrosis has raised new questions with regard to the need of management and the indications for operation. Moreover, there is considerable controversy whether or not pyeloplasty improves long-term renal function. We evaluated the operative results of this entity under the setting of well defined indications for surgery. MATERIALS AND METHODS: The indications for pyeloplasty included Society for Fetal Urology-grade (SFU-grade) 3 and grade 4 hydronephrosis on ultrasonogram together with obstructive pattern on modified "Well Tempered" diuretic renogram. We analysed the postoperative changes of ultrasonographic findings, differential renal function, and renogram patterns in 28 children with unilateral hydronephrosis detected prenatally. RESULTS: Of the patients, 27 had grade-4 hydronephrosis on preoperative ultrasonogram. Pyeloplasty was effective in decreasing the grade of dilatation in all kidneys except one. Of the involved renal units, 14 (50%) had reduced differential renal function less than 45% on the preoperative renogram. Differential renal function increased significantly by absolute value of 5% or greater in 8 kidneys (29%), but decreases in 5 kidneys (18%). In the remaining 15 kidneys (54%) there was no significant differences between pre- and post operative differential renal function. Postoperative diuretic renogram changed to non-obstructive pattern in 26 kidneys (93%). Another 2 kidneys showed hypofunctioning pattern. CONCLUSIONS: The results suggest that, although pyeloplasty decreases ultrasonographic dilatation of hydronephrosis and improves excretory pattern of diuretic renogram, significant increase of differential renal function is expected in only one third of the involved kidneys. PMID- 10826247 TI - [Clinical evaluation of low dose glucocorticoid therapy for hormone-refractory prostate cancer]. AB - Fifteen cases with hormone-refractory metastatic carcinoma of the prostate were treated by the combination therapy of LH-RH analogue and low-dose glucocorticoid. Prior treatments of the patients were composed of maximum androgen blockade (MAB) therapy (14 cases) and bilateral orichiectomy (1 case). Nine of 15 cases underwent estramustine based treatment or anti-cancer chemotherapy as the second line therapy. As a glucocorticoid (steroid) therapy, 1.5 or 2.0 mg of dexamethasone or 10 mg of predonisolone were given for a median period of nine months. Clinical responses were evaluated by the determination of serum PSA, EOD score and QOL scales. PSA responses were classified as CR, PR and PD according to the following categories; CR : PSA level decreased to less than 2 ng/ml, PR : PSA level decreased to less than half values of pretreatment level, PD: 20% or more increased to pretreatment level. PSA responses (CR + PR) were observed in eight of 15 patients for a median period of six months. Responses are seemed to be correlated with the Gleason scores. No apparent improvement on EOD scores by bone scintigraphy was seen, however, definite improvement of bone pain was identified in eight of 11 cases. The steroid therapy was considered as one of the useful treatment choice for patients with hormone-refractory prostate cancer. PMID- 10826248 TI - [A case of neuroendocrine tumor of the urinary bladder]. AB - We present a case of a neuroendocrine tumor of the urinary bladder. A 81-year-old man presented with macrohematuria and difficult voiding. Cystoscopy, computed tomography (CT) and magnetic resonance imaging (MRI) revealed non-papillary broad based bladder tumor, benign prostatic hyperplasia and three bladder stones. Histological diagnosis was neuroendocrine tumor of the urinary bladder and he underwent two courses of trans-arterial infusion (neoadjuvant) chemotherapy. After two courses of chemotherapy, pelvic CT scan revealed partial response and then the patient underwent transurethral resection of the bladder tumor, simultaneously with TUR-P and cystolithotripsy. Examination of the resected specimen indicated pathological complete remission and he discharged. Although the patient was followed-up postoperatively because of pathological complete response, the tumor recurred in the bladder three months later. PMID- 10826249 TI - [A case of metanephric adenoma treated with partial nephrectomy]. AB - A 32-year-old female presented with a right renal mass detected by abdominal ultrasound examination at a routine medical check up. Computed tomography scans and magnetic resonance images revealed a solid tumor in the upper pole of the right kidney measuring 1.5 cm in diameter. Partial nephrectomy was performed on suspicion of the renal cell carcinoma. Histologically, the tumor is composed of small, uniform, epithelial cells with scant cytoplasm and hyperchromatic round nuclei that formed a tubular or glomerular-like structure. The histological diagnosis was metanephric adenoma. PMID- 10826250 TI - [Management of unilateral renal cystine calculi via percutaneous nephrolithotripsy in a child with a contralateral non-functional kidney--a case report]. AB - A 2-year old boy visited our clinic with a chief complaint of high fever. A past history of acute renal failure due to cystine stones and cystinuria was expressed. Abdominal rentogenograms and CT demonstrated a right ureteral stone and a left renal stone. Furthermore renogram evaluation indicated non-function of the right kidney and dysfunction in the left kidney. Since right ureteral stone moved into bladder seven days post-admission, right ureteroscopy, left PNL, and cystolithotripsy were performed. Considering that right ureteral stenosis was determined by ureteroscopy, balloon dilation against the stenotic ureteral wall was performed. Left PNL and cystolithotripsy were successfully performed. No intraoperative complications occurred and no symptoms of signs of recurrence of the underlying metabolic disease were evident four months postoperatively. PMID- 10826251 TI - Women's responses to sexual violence by male intimates. AB - The purpose of this grounded theory study was to devise a theoretical framework that describes the problem of sexual violence by male intimates from the point of view of 23 women who have experienced such violence at some time in their adult lives. The core variable, forging ahead in a dangerous world, reflects the women's descriptions of life after violence as a struggle to get on with their lives in a social world they know through first-hand experience to be unsafe. The theoretical framework includes three variations of forging ahead (getting back on track, starting over again, and surviving the long, hard road) described by three subgroups of women who experienced different types of sexual violence. The framework also outlines three common processes used to forge ahead: telling others, making sense of the violence, and creating a safer life. The nature and meaning of these processes differ according to group. PMID- 10826252 TI - Personality traits and self-care in adults awaiting renal transplant. AB - This article reports the pretransplant findings of the first phase of a three phase, longitudinal study examining relationships among personality traits and self-care abilities and behaviors of Ontario adults pre- and post-renal transplant. A consortium of Ontario nurse researchers representing three of Ontario's five renal transplant centers conducted this research. All adults on the cadaver transplant lists of 15 Ontario dialysis centers were invited to participate. One hundred ninety-eight adults awaiting renal transplant were enrolled in the study, representing a 70% response rate. A cross-sectional, correlational design was used for the pretransplant phase. Self-report measures with known psychometric properties were used; validity and reliability of the measures were supported by the sample. Data were analyzed using descriptive approaches, correlational analyses, multiple regression, and path analysis. Relationships were supported among selected personality traits, health state and self-care abilities and behaviors. Further research to examine personality traits and health state in relation to adult self-care is warranted. PMID- 10826253 TI - Men's perspectives on individual and family coping with their wives' breast cancer and chemotherapy. AB - Little research has examined the impact of cancer and chemotherapy treatment for breast cancer from men's perspectives as partners, fathers, and caregivers. This research, part of a larger study describing women's, partners', and children's perspectives, aims to describe men's perspectives on their experiences and how their wives' breast cancer and chemotherapy impacted them and their families, to describe what facilitated and hindered their coping, and to suggest interventions to assist men and their families to manage the experience with less stress. This participatory action study used qualitative naturalistic inquiry methods. Semistructured interviews were conducted with 11 male partners. Two major themes were identified: focusing on a wife's illness and care, and focusing on the family to keep life going. Nine sub-themes cut across the major themes: being there, relying on health care professionals, being informed and contributing to decision making, trying to keep patterns normal and family life going, helping out and relying on others, being positive, putting self on hold, adapting work life, and managing finances. PMID- 10826254 TI - Cost-effectiveness analysis methods for the REACT Study. AB - The Reconditioning Exercise and Chronic Obstructive Pulmonary Disease Trial (REACT) is a two-arm randomized clinical trial designed to compare short-term versus long-term exercise intervention in terms of physical function, acute exacerbation of chronic obstructive pulmonary disease, health-related quality of life, and cost-effectiveness. Clinical trials such as REACT are now routinely paired with economic analyses, and nurses can expect to play a growing role in the conduct of these studies. This article describes a model that is useful for structuring economic evaluations of health care interventions, and illustrates a cost-effectiveness analysis that is being conducted in conjunction with the REACT study. An in-depth description of collection methods and procedures is provided, as well as a summary of recruitment and retention experience to date. PMID- 10826255 TI - Social and professional influences of the technology of electronic fetal monitoring on obstetrical nursing. AB - Electronic fetal monitoring (EFM) is one example of a biomedical technology that rapidly diffused from an experimental innovation into a standard medical practice. First developed in the 1950s, EFM became commercially available in the early 1970s and quickly transformed intrapartum obstetrical practice. Assessments and interventions, which practitioners had previously based primarily on laboring women's subjective reports of bodily sensations, were now being based on quantifiable objective data from uterine activity and fetal heart rate transducers. Despite concerns of over-medicalization of the natural event of birth, iatrogenesis related to the increased incidence of operative deliveries, and escalating costs, EFM became widely accepted as routine and necessary by both practitioners and patients. By presenting the confident expectations and cautious reservations of various practitioners and patients to EFM, this article explores the rapid diffusion of EFM within the social context of the 1970s. A special focus is given to the perspective of intrapartum obstetrical nurses, because they have been the primary users of this perinatal technology since its introduction. PMID- 10826256 TI - Nursing advocacy during a military operation. AB - Advocacy is an essential component of the registered nurse's professional role, yet experts provide no consistent definition of advocacy. The purpose of this study was to explore the experiences of military nurses as they engage in advocating practices and to describe their shared practices and common meanings. Heideggerian hermeneutic phenomenology, provided the framework and method for this study. Twenty-four U.S. Army nurses were individually interviewed and the researcher kept interview observational notes. The constant comparative method of analysis was used. The stories of these nurses revealed one constitutive pattern- safeguarding--and four related themes. The themes were advocating as protecting, advocating as attending the whole person, advocating as being the patient's voice, and advocating as preserving personhood. One conclusion was that military nurses must be prepared for the important safe-guarding role. They must be coached in how to deal with other members of the health team on the patient's behalf. PMID- 10826257 TI - Informed consent and the limits of confidentiality. AB - Investigators encounter many legal and ethical issues when they conduct research on partner abuse. Balancing abused women's rights to privacy with legally mandated reporting requirements involves considerable thought and planning. Failure to protect participants may result in escalation of abuse as well as loss of children to protective services. Various perspectives on maintaining participant privacy while conforming to legal mandates to report child abuse, homicidality, and suicidality are discussed. The role of confidentiality certificates in providing legal immunity for researchers and the method of obtaining the certificates are presented. In addition, the authors describe the strategies for participant protection that are implemented in a federally funded study of abuse, women's self-care, and pregnancy outcomes. The decision to clearly and specifically inform abused women of the limits of confidentiality allows participants to make informed decisions about disclosures, but may result in diminished recruitment. PMID- 10826258 TI - The realities of aging. PMID- 10826259 TI - Supplemental vitamin C may hasten atherosclerosis. PMID- 10826260 TI - Geriatrics photo quiz. Galactorrhea. PMID- 10826261 TI - Pruritic foot lesions. Summertime heat and sweat exacerbate the condition. PMID- 10826262 TI - Hypercholesterolemia. Is lipid-lowering worthwhile for older patients? AB - Despite some indications to the contrary, evidence continues to accumulate that controlling cholesterol levels with drug therapy in older persons is a worthwhile goal. Older persons with hypercholesterolemia have an elevated risk of coronary heart disease morbidity and mortality, and this risk increases as they age. Recent clinical trials have suggested that older persons benefit from lipid lowering therapy as much as younger patients do. Therefore, intervention appears to be justified--as is greater vigilance in identifying untreated patients. PMID- 10826263 TI - Osteoporosis. An overview of the National Osteoporosis Foundation clinical practice guide. AB - During the past decade, numerous organizations and associations have published recommendations for the prevention and treatment of osteoporosis. For the primary care physician, the most applicable of these--due to its reliance on clinical trial data and its scope--is the clinical guide published by the National Osteoporosis Foundation. The guide addresses risk assessment, bone mineral density testing, diagnosis, nutritional supplementation, and pharmacologic therapy, including consideration of the newer agents used to slow or manage osteoporosis progression. Reflecting one of the key deficiencies in the clinical trial data, the guide applies predominantly to a patient population of postmenopausal white females. The refined design of new osteoporosis studies will in time allow for recommendations that apply to a more diverse patient population. PMID- 10826264 TI - Mycotic infections. Effective management of conditions involving the skin, hair, and nails. AB - Superficial mycotic infections of the skin, hair, or nails are recurring presentations in the geriatric primary care setting. The most common infections are those caused by dermatophytes. The genus Trichophyton gives rise to most of the tinea dermatophytoses, including tinea capitis, tinea pedis, and tinea unguium (onychomycosis). Part of the diagnostic challenge lies in distinguishing the mycotic lesions from those caused by cutaneous diseases such as psoriasis, eczema, dyshidrosis, and contact dermatitis. Because environmental conditions play a major role in fungal infection onset, clinical management should include patient education about conditions conducive to fungal propagation. Oral agents are the primary mode of treatment for fungal infections of the scalp and nails, whereas topical treatments are frontline agents for other superficial skin conditions. PMID- 10826265 TI - Breathlessness in spinal cord injury depends on injury level. AB - Little is known about the prevalence and predictors of breathlessness in individuals with neurologically complete chronic spinal cord injury (SCI). Between December 1992 and September 1993, we mailed a respiratory questionnaire to 1,147 community-based individuals with chronic SCI. The questionnaire included four questions about the presence of breathlessness during activities related to moving about. Of the 485 who replied (42 percent response rate), analysis was limited to adult males with neurologically complete motor injuries who reported using a hand-propelled wheelchair more than 50 percent of the time to get around. Of 130 subjects (33 tetraplegics, 53 high thoracic SCI, 44 lower injury levels), the patients with tetraplegia reported breathlessness more frequently (range for the four questions, 21-33%) than those with high thoracic (range, 9-15%) or lower injury levels (range, 2-11%). For each of the four questions there was a significant trend (p < 0.05) for subjects with higher levels of injury to report the greatest prevalence of breathlessness (tetraplegia > high thoracic > lower). The frequency of breathlessness was greatest in those with neurologically complete cervical injuries, an effect that was independent of obesity, smoking, age, and years since SCI. The mechanisms of breathlessness in SCI are unclear but elucidation might lead to strategies for providing relief. PMID- 10826266 TI - Epididymal sperm transport in normal and recent spinal cord injured Sprague Dawley rats. AB - Causes of poor semen quality following spinal cord injury (SCI) are not known. One possible reason, based upon studies that reported improved semen quality in SCI men after several induced ejaculations, is delayed epididymal sperm transport. Our study was designed to establish baseline epididymal sperm transport values in the Sprague Dawley rat and evaluate effects of SCI on this process. Spermatozoa protamine was labeled with tritiated arginine, and the rats were sacrificed various times after injection. Each epididymis was divided into six equal sections from proximal to distal. Sperm tails were dissolved with 8 molar (M) urea in the presence of 2 mM dithiothreitol (DTT); sperm heads were collected by centrifugation (3,000 rpms, 10 min.). The radioactivity in sperm heads from each section was counted and expressed as counts per million sperm heads. To account for different rates of labeled arginine incorporation, the percentage of counts per million sperm heads in each section was calculated relative to the total number of counts in all six sections. Our results showed there was an orderly progression of sperm through the epididymis. It took 8 days for labeled sperm to enter the epididymis and 28 days to peak in the caudal (tail) section in non-SCI rats. Stasis was present 10 days after T-9 SCI in rats compared with transport in sham controls. This was evidenced by a significant increase in the percentage of labeled sperm in proximal sections of the epididymis (sections 1, 2, and 4) in T-9 transected animals (p < 0.01). If similar stasis occurs in SCI men, it could obviously contribute to poor semen quality. However, it remains to be determined how long this stasis persists after SCI in rats. PMID- 10826267 TI - Outcome of functional electrical stimulation in the rehabilitation of a child with C-5 tetraplegia. AB - Hand function was provided for a six-year-old child with C-5 American Spinal Injuries Association (ASIA) classification-A tetraplegia through a percutaneous intramuscular (i.m.) functional electrical stimulation (FES) system. In conjunction with implantation of 10 percutaneous i.m. electrodes for provision of grasp and release of her right hand, reconstructive surgery was performed to provide upper extremity positioning to optimize hand use. The subject participated in FES training over a nine-week period for approximately five hours weekly, with an additional five hours each week dedicated to exercise and conditioning of her arm muscles. Physical and functional assessments included range of motion (ROM), manual muscle testing (MMT), activities of daily living (ADL) abilities and the Canadian Occupational Performance Measure (COPM), used to evaluate the effect of stimulated hand function and surgical reconstruction on functional ability. These were conducted prior to FES and surgery and repeated after rehabilitation training. With rehabilitation and training, the child was able to control her FES system. Physical assessments revealed increased strength of both shoulders and more useful range of arm movement. Functional assessments show that the FES system enabled her to perform age-appropriate ADL that previously were achievable only with physical assistance. Her overall level of independence in ADL ability increased, as did self-rated levels of satisfaction and performance on chosen activities. Positive gains demonstrated here suggest the need for further studies of FES systems in young children with SCI. PMID- 10826268 TI - Intravesical capsaicin therapy: a review. PMID- 10826269 TI - A review of the neuropathology of human spinal cord injury with emphasis on special features. AB - A judicious understanding of the basic neuropathology of spinal cord injuries (SCI) is essential knowledge for the clinician responsible for SCI management. An appreciation of the nature of human SCI is also necessary for the neuroscientist searching for a cure. The neuropathology of human SCI described here is derived from the study of 564 cases of spinal cord trauma held in a tissue bank and database of the Department of Neuropathology, Royal Perth Hospital in Australia. The main features of SCI neuropathology are reviewed and special aspects such as early axonal lesions, traumatic demyelination-remyelination, and quantification of white matter tracts are reported in more detail. One of the remarkable outcomes of this work is the finding that the majority of SCI patients have a proportion of spinal cord white matter maintained across the level of the lesion, an observation that has important therapeutic implications. PMID- 10826270 TI - Methicillin-resistant Staphylococcus aureus in patients with spinal cord injury. AB - During the past decade, methicillin-resistant Staphylococcus aureus (MRSA) has become a major cause of nosocomial and community-acquired infections in the United States. A retrospective chart review of MRSA-positive cases from January 1990 to December 1994 was done to assess the extent of the problem, the factors associated with MRSA acquisition, and the effectiveness of a Center for Disease Control (CDC) protocol to treat MRSA infection in a population with spinal cord injury (SCI). Seventy-four percent of the patients acquired MRSA during their hospitalization. Urine was the most common site of colonization, and 73 percent of the patients with positive urine cultures managed their bladders with indwelling catheters. Implementation of the CDC protocol in 1991 was associated with a decrease in the MRSA incidence rate in subsequent years. Other simple methods of treatment, such as bladder irrigation, were also effective. To control the spread of MRSA, a continual concerted effort by hospital staff through education and implementation of the MRSA protocol is necessary. PMID- 10826271 TI - APS recommendations for skin care of hospitalized patients with acute spinal cord injury. PMID- 10826272 TI - APS recommendations for the urological evaluation of patients with spinal cord injury. PMID- 10826273 TI - Recovery of locomotion after experimental spinal cord injury: axonal regeneration or modulation of intrinsic spinal cord walking circuitry? PMID- 10826274 TI - Managed care and SCI. PMID- 10826275 TI - Putting the lid back on Pandora's jar. PMID- 10826276 TI - Source Informatics Ltd versus Department of Health: appeal court reverses judgement of Mr Justice Latham and allows use of anonymized patient information without consent. PMID- 10826277 TI - Medical errors major killers in UK and USA. PMID- 10826278 TI - Famous names in toxicology. Inoculators and vaccinators of the 18th century III. PMID- 10826279 TI - Aggression as an adverse drug reaction. PMID- 10826280 TI - Drug interactions with drugs used in the treatment of HIV and infective complications peculiar to HIV infected patients. PMID- 10826281 TI - The century's greatest contributions to control practice PMID- 10826282 TI - Modeling of an intelligent pressure sensor using functional link artificial neural networks. AB - A capacitor pressure sensor (CPS) is modeled for accurate readout of applied pressure using a novel artificial neural network (ANN). The proposed functional link ANN (FLANN) is a computationally efficient nonlinear network and is capable of complex nonlinear mapping between its input and output pattern space. The nonlinearity is introduced into the FLANN by passing the input pattern through a functional expansion unit. Three different polynomials such as, Chebyschev, Legendre and power series have been employed in the FLANN. The FLANN offers computational advantage over a multilayer perceptron (MLP) for similar performance in modeling of the CPS. The prime aim of the present paper is to develop an intelligent model of the CPS involving less computational complexity, so that its implementation can be economical and robust. It is shown that, over a wide temperature variation ranging from -50 to 150 degrees C, the maximum error of estimation of pressure remains within +/- 3%. With the help of computer simulation, the performance of the three types of FLANN models has been compared to that of an MLP based model. PMID- 10826283 TI - Accurate predictive modeling of response variables under dynamic condition without the use of past response data AB - One promising attribute of the dynamic predictive modeling method introduced by Rollins et al. [D.K. Rollins, J. Liang, P. Smith, Accurate simplistic predictive modeling of nonlinear dynamic processes, ISA Transactions 37(4) (1998) 193-203] is its ability to accurately predict output response without the use of online output data. The proposed method only needs online input data to accurately predict output behavior once the semi-empirical model has been identified using offline data. This ability is critical to chemical processes because many output variables (such as chemical composition) are often measured infrequently, inaccurately, or not at all. In addition, in the presence of extremely high measurement noise of the output variable, this work will demonstrate very accurate predictive performance. Finally, this article will show that the method of Rollins et al. can predict better without the use of output data than with the use of output data in the case of large measurement variance. Thus, the proposed method is being recommended for its accuracy, especially in situations where online output response data is limited or inaccurate. PMID- 10826284 TI - Data reconciliation with uncertain models AB - Data reconciliation has proven to be an effective technique for providing frequent, accurate and consistent "best estimates" of plant operation data. However, in almost all the proposed techniques until today, the mathematical model of the process has been considered as exact. In point of fact, this hypothesis is uncommon and frequently the models used are uncertain. This paper proposes a new technique of data reconciliation which is able to exploit the knowledge about the uncertainties of the model with regard to which the reconciliation is done. It leads to the solution of a classical quadratic optimisation problem subject to constraints. The originality of the proposed technique is to use penalty functions for solving this problem and to weight each constraint with regard to their uncertainties. PMID- 10826285 TI - Innovation sequence application to aircraft sensor fault detection: comparison of checking covariance matrix algorithms AB - In this paper, the algorithms verifying the covariance matrix of the Kalman filter innovation sequence are compared with respect to detected minimum fault rate and detection time. Four algorithms are dealt with; the algorithm verifying the trace of the covariance matrix of the innovation sequence, the algorithm verifying the sum of all elements of the inverse covariance matrix of the innovation sequence, the optimal algorithm verifying the ratio of two quadratic forms of which matrices are theoretic and selected covariance matrices of Kalman filter innovation sequence, and the algorithm verifying the generalized variance of the covariance matrix of the innovation sequence. The algorithms are implemented for longitudinal dynamics of an aircraft to detect sensor faults, and some suggestions are given on the use of the algorithms in flight control systems. PMID- 10826286 TI - Development of a GPC-based PID controller for unstable systems with deadtime AB - Till now, traditional low-order control schemes have never been applicable to unstable systems with deadtime. In this paper, we present the first application of a PID controller with time-scheduled gains to unstable systems with deadtime, consisting of a single unstable pole. The control gains are designed based on a generalised predictive control (GPC) approach. The only user specifications required are simple and classical desired properties as in the natural frequency and the damping ratio of the closed-loop system. An approach is further developed to subsequent on-line self-tuning of control weights so that the overall control system remains applicable and effective in the face of disturbances and slowly varying dynamics. A detailed analysis of the closed-loop stability of the thus designed control system is further provided in the paper. Based on stability conditions developed, the prediction horizon for the GPC-based controller may be effectively computed. Finally, simulation examples illustrate the performance of the control system. PMID- 10826287 TI - An adaptive I-PD controller based on frequency domain system identification AB - This paper presents an approach to auto-tune an I-PD controller for plants with long time delays such as pneumatic cylinders. The proposed control scheme estimates the frequency response of a plant using the sliding discrete Fourier transform (DFT) and employs a modified Smith predictor to reduce the effect of time delay. Controller gains are determined so as to minimize the error between the open-loop frequency response of the reference model and that of the actual system. PMID- 10826288 TI - Re-design of Smith predictor systems for performance enhancement AB - In this paper, a new design for Smith Predictor systems is presented. It employs a deliberately mismatched model to enhance performance over a perfectly matched system while using a simple primary controller. The design methodology is formulated in the frequency domain as an optimization problem and it turns out that an approximate solution can be obtained using the linear least squares method. To improve the performance further, a modified Smith Predictor system structure is also proposed, and it reduces the system into one involving second order dynamics for the primary controller design. Illustrative simulation of several typical processes are included. PMID- 10826289 TI - Real-time flow control using neural networks AB - In this paper, an on-line trained neural network controller is applied to control the flow rate of a process control rig. The neural controller replaces a conventional controller in the forward path. The overall performance of this controller is compared with that of a PID controller in the presence of noise and non-linearity. It is shown that as the non-linearity is added to the system, the PID controller cannot track the set-point changes, however, the neural controller copes well under various conditions. PMID- 10826290 TI - Advantages of an electrical control and energy management system AB - This paper discusses an electrical control and energy management system (ECEMS) that was installed at Indian Petrochemicals Corporation Limited (IPCL) Nagathone Gas Cracker complex located in Maharashtra, India. This distributed control system (DCS) provided computer assisted control in the areas of: Demand control; Automatic generation control, including MW and MVAR management; Power factor control; Automatic tap changer control; Load shedding; Automatic synchronization of generator and ties; Remote control of breakers. Previously, IPCL, like most other petrochemical companies in India, relied on operator control for power house functions. The process is always automated, but the power house equipment is usually manually controlled. Electrical control and energy management systems are not thought to be necessary. However, in this case the consultants for IPCL and the DCS supplier convinced IPCL that an ECEMS would save them enough money in operating costs to pay for the new control system. The control system discussed in this paper reduced operating costs by satisfying the process steam and power demands in the most cost-effective manner. In addition, the system took action to respond to electrical disturbances, such as loss of tie line and generator tripping, so that stable conditions were restored. PMID- 10826291 TI - Process control reliability: the key to investing in your infrastructure AB - Expenditures to improve process control infrastructure in pulp and paper mills should be analyzed and evaluated as rigorously as greenfield investments to optimize capital effectiveness. An approach to such an evaluation based on the economic impact of process control equipment reliability is presented. The procedure, called the control infrastructure health assessment (CIHA), is described and the results obtained for a particular case are discussed. Some data and the linkage to life cycle costing (LCC) are shared. PMID- 10826292 TI - Medial temporal lobe amnesia: a case study for nursing. AB - Maintaining and enhancing cognitive function is a crucial but challenging intervention for patients with memory problems. Research on the medial temporal lobe (MTL) memory system has yielded much information that can guide nurses in planning, evaluating, and performing effective interventions. A patient, Mrs. N, with a diagnosis of anaplastic astrocytoma of the left medial temporal lobe provides an example. Information from research guides assessment of Mrs. N and affords development of specific patient-centered interventions to maintain function, cope, and compensate. Data have been gathered from the patient, relatives, and caregivers to compare with and augment existing research, because few nursing case studies of amnesia involving patients with left medial temporal lobe tumors are available for analysis. PMID- 10826293 TI - A follow-up study of 14 young adults with complex regional pain syndrome type I. AB - The purpose of this study was to evaluate the present symptom experience and functional status of 27 young adults with Reflex Sympathetic Dystrophy Syndrome, now known as Complex Regional Pain Syndrome (CRPS) type I. All were originally diagnosed and studied more than 12 years ago. Only 15 subjects could be located; 14 participated in this research. A descriptive exploratory research design was utilized. A tool was designed to gather data about daily living variables, psychosocial variables, and symptom variables. Although many questions required the use of Likert scales, there were some open-ended questions to capture the qualitative descriptions of the participants' symptom experiences. Findings contribute to the clinical literature about CRPS and its effects on clients over time. PMID- 10826294 TI - Functional hemispherectomy: radical treatment for Rasmussen's encephalitis. AB - Rasmussen's encephalitis is a progressive, debilitating disease that usually appears in the pediatric population but also may affect adults. Uncontrolled seizures, cognitive decline, and a progressive hemiparesis accompany this disorder. Treatment options are limited and consist of medical management, which is somewhat experimental and ineffective; the use of either intravenous antiviral agents or steroids; or surgical resection of the affected hemisphere. Patients undergoing hemispherectomy for Rasmussen's encephalitis are complex. Nurses caring for these patients require a high degree of knowledge and specialized skills. Patient outcome is highly affected by the knowledge and skill of the entire team of healthcare professionals needed to manage these patients. PMID- 10826295 TI - Stress, depression, and psychoneuroimmunology. AB - Stress is a common phenomenon most people encounter every day, whereas depression represents a vast constellation of mental disorders of multifarious etiology. Both situations have been exhaustively evaluated and scrutinized in the complex field of psychoneuroimmunology (PNI). PNI studies have attempted to ascertain and define the extensive links among and intricate web of neural activity, the endocrine system, and altered immune responses in stressed patients and those with depressive disorders. Although the literature indicates some type of relationship exists between these systems, the associations between and relevance of immune alterations in depression and stress to health and illness remain to be determined. Only by seeking to expand our knowledge of these neurological, endocrine, and immunological alterations in depressed and stressed neurological patients will our understanding of stress, depression, and health be magnified and our ability to provide holistic care be maximized. PMID- 10826296 TI - Evaluation of an algorithm for treatment of status epilepticus in adult patients undergoing video/EEG monitoring. AB - Convulsive or generalized tonic clonic status epilepticus (SE) is a neurological emergency that can lead to transient or permanent brain damage or even death. An algorithm was designed to aid nursing and medical staff members in decision making about the type of SE and pharmacological intervention needed to stop prolonged or repetitive seizures. Fifteen registered nurses at a northern New England medical center's epilepsy unit participated in educational sessions on classification of seizures and status epilepticus prior to use of the algorithm. A pretest-posttest design with an investigator-developed tool was used to measure SE knowledge before and after educational intervention. There was a significant improvement in scores on the posttest of the classification of status epilepticus (Z = -2.93, p = .003). Twenty-nine medical records of patients who had experienced SE between February 1992 and December 1997 were reviewed. Nineteen patients experienced SE before the algorithm was implemented, and 10 patients experienced SE after the algorithm was implemented. A total of 16 patients experienced generalized convulsive SE with 12 episodes occurring before and 4 episodes after algorithm implementation. The mean time taken to stop the episode of SE after pharmacologic treatment began was compared in both groups using a t test. The mean difference between the groups was 235 minutes (t = 2.57, p = .026). The findings of this project demonstrate that combining a treatment algorithm with education of staff members on its use has benefits in the practice setting of an inpatient comprehensive epilepsy program. Episodes of SE are more accurately classified and successful treatment of the episodes occurs earlier. PMID- 10826297 TI - The heart-brain connection. AB - We have long known that patients with vascular disease in one system are at risk for vascular disease in other systems. Beyond this, we are recognizing the increased risk for cardiovascular patients to develop stroke not only as the result of arrhythmia, but also at the time of cardiovascular events or procedures. This presents clinical challenges to nurses with either neurological or cardiovascular expertise, requiring development of new awareness, clinical and critical thinking skills, and collaboration with their colleagues in other specialties. Three case studies illustrate patient presentations ranging from the subtle to the obvious. Pathophysiology of stroke is reviewed. Leading-edge management strategies and supporting literature highlight the benefits of prompt identification and management of the stroke patient. The Stroke Watch Action Team (SWAT) has proved to be an effective means of expediting patient identification and access to effective stroke treatment. PMID- 10826298 TI - Neurobiological bases for Alzheimer's disease. AB - The ever-increasing number of persons with Alzheimer's disease (AD) will create a greater demand for neuroscience nurses in the 21st century. There are approximately 4 million Americans with AD, and this number is expected to increase to approximately 15 million by the year 2050. As neuroscience nurses encounter more patients with AD in their daily practice, a better understanding of this neurodegenerative disease is important in order to provide high-quality nursing care. An insight into the neurologic bases for cognitive and behavioral manifestations of AD enables nurses to implement effective nursing interventions. PMID- 10826299 TI - Should boxing be banned? AB - Should boxing be banned? Do the ever-so-obvious risks outweigh everyone's freedom to choose whether to expose oneself to these risks by taking up the sport? On an official level, the RCN in the UK has taken its stand--it does not! So has also the British Medical Association (BMA)--it does! With few exceptions, the responding nurses from Europe, America, and Australia in this month's column seem to agree with the official nursing standpoint in the UK, also emphasizing the importance that any person's choice not only should be free, but also informed. In the United States, where boxing perhaps has its strongest tradition and deepest roots, the whole issue hardly seems to be one of much realistic debate at all. In Australia, however, the debate seems to be similar to that in the UK. What would a total ban on boxing lead to? No more boxing and no more neurological consequences due to boxing? Doubtfully, boxing would probably continue anywhere where there is an interest for it, and a ban might actually increase the attraction to the sport for some people. In this scenario there is also a risk that the safety precautions would be seriously compromised. This month's question exemplifies an area in which it is very important for nurses to make a stand, on a personal as well as on a collective level. As indicated by several of this month's replies, the issue is probably not merely about boxing but also about to what extent people's choices should be controlled by bans and where the line should be drawn. To what extent are people competent to make their own decisions and where/when/how should "big brother" (in this case as represented by, among others, nursing as a profession) be allowed to step in? Anyone who has any further contributions or comments on this issue is welcome to contact me! PMID- 10826300 TI - Heterosexual prevention: the key to averting HIV orphanhood. PMID- 10826301 TI - Predictors of perception of cognitive functioning in HIV/AIDS. AB - This is a descriptive, correlational study of the predictors of perceived cognitive functioning. The convenience sample of 728 nonhospitalized persons receiving health care for HIV/AIDS was recruited from seven sites in the United States. All measures were self-reported. Self-perception of cognitive functioning, the dependent variable, was composed of three items from the Medical Outcomes Study HIV scale: thinking, attention, and forgetfulness. Data related to age, gender, ethnicity, education, injection drug use, CD4 count, and length of time known to be HIV-positive were collected on a demographic questionnaire. The scale from the Sign and Symptom Checklist for Persons with HIV Disease was used to measure self-reported symptoms. Data were analyzed using hierarchical multiple regression analysis. Predictors of perception of cognitive functioning explained a total of 36.3% of the variance. Four blocks--person variables (1.5%) (age, gender, education, history of injection drug use), disease status (2.3%), symptom status (26.5%), and functional status (5.4%)--significantly contributed statistically to the total variance. Among those individuals who completed the questions related to depression (n = 450), 28% of the variance in cognitive functioning was explained by this variable. The findings in this multi-site study indicate that symptom status explained the largest amount of variance in perceived cognitive functioning. Early identification of cognitive impairment can result in appropriate clinical interventions in remediable conditions and in the improvement of quality of life. PMID- 10826302 TI - Sources of occupational stress and coping strategies among nurses working in AIDS care. AB - Nurses working in AIDS care experience high rates of occupational stress and therefore are vulnerable to emotional exhaustion and occupational burnout. This study surveyed 499 members of the Association of Nurses in AIDS Care regarding their work-related stress experiences and coping strategies for managing stress. Qualitative analyses identified a hierarchical structure of occupational stress, with two supraclusters representing workplace and patient care-related stress and eight specific subclusters of stressors: institutions, personnel, biohazards, death, informing patients, challenging patients, families, and treatment dilemmas. Analyses showed that nurses experiencing stress from their workplace were significantly more likely to use wishful thinking, planful problem solving, and avoidance as coping strategies, whereas stress originating from patient care was more likely to be dealt with using positive appraisal and acceptance. Interventions designed to assist nurses in managing occupational stress and to prevent occupational burnout must include the sources of work-related stress among nurses in AIDS care. PMID- 10826303 TI - Identifying HIV/AIDS research priorities for the next millennium: a Delphi study with nurses in AIDS care. AB - The purpose of this study was to identify HIV/AIDS research priorities for the next millennium from the perspective of nurses in AIDS care. The study used a three-round Delphi technique, with study participants (the panel of experts) selected from the membership of the Association of Nurses in AIDS Care. In the first round of the Delphi study, 317 nurses identified more than 2,000 topics they viewed as important for overall HIV/AIDS research and for HIV/AIDS nursing research. Using qualitative analysis, 12 priority topics were generated in the overall research category and 12 priority topics in the nursing research category. In Round II of the Delphi survey, study participants ranked the 12 topics in categories (overall and nursing research priorities) identified in Round I. Finally, in Round III, the top five priority topics from Round II were prioritized in each category. The overall HIV/AIDS research topics identified were (a) HIV community-level education and prevention; (b) development of more tolerable drugs; (c) HIV prevention focusing on individual or specific group behavior; (d) vaccine development; and (e) development of new and more effective drugs. The HIV/AIDS nursing research priorities identified were (a) symptom management; (b) community-level HIV education and prevention; (c) quality of life issues in chronic HIV disease; (d) HIV prevention focusing on individual or specific group behavior; and (e) research related to adherence to drug therapy. PMID- 10826304 TI - Active client participation: an examination of self-empowerment in HIV/AIDS case management with women. AB - Case management has become increasingly popular as a means to alleviate the difficulties associated with accessing health care and social services. However, little information exists regarding models of case management specifically intended for work with HIV-positive women. This article explores the practices currently in use at an HIV/AIDS service agency to further define empowering practices employed by case managers working with HIV-positive women. Although a client's active participation in service plan development and delivery is widely regarded as the pathway to empowerment, findings indicate that "active participation" is not so readily defined; empowerment exists on a dynamic continuum with increasing levels of client participation. A model is presented that demonstrates the relationship of particular behaviors to client self empowerment. Because the needs of clients with HIV/AIDS are continually fluctuating, and because case managers' behaviors will need to adapt accordingly, this model also incorporates the concept of balance as it pertains to HIV/AIDS case management. PMID- 10826305 TI - Developing culture-specific interventions for Latinas to reduce HIV high-risk behaviors. AB - Latina women are among the fastest-growing segments of the U.S. population to become infected with HIV. Health care providers must use culture-specific HIV prevention strategies and must address cultural roles and values of the Latin community to empower Latinas to reduce high-risk behaviors. Some Latinas adhere to their cultural traditions that place the female subservient to the male in the sexual relationship. A review of the literature reveals that skills-building, a group participatory tool that engages the individual to examine high-risk behaviors and provides methods to make behavior changes, and nurse counseling, which focuses on an individual's specific risk behaviors, are more effective modes of reducing high-risk behaviors in women than an information-only intervention. PMID- 10826306 TI - The lived experience of grieving for persons living with HIV who have used injection drugs. AB - Parse's research method was used to investigate the lived experience of grieving for 10 persons self-identified as HIV-positive injection drug users. These individuals compose an understudied and poorly understood population, and their grief experiences have rarely been documented. The losses grieved by persons living with HIV infection include the loss of life, friends, family members, employment, energy, and sex. The lived experience of grieving was found to be "overwhelming anguish that shapes hopes and intentions as a wretched aloneness is punctuated with cherished uplifting engagements, while gratitude inspires courage in the midst of ambiguity." This new conceptualization of the grieving process is discussed in light of Parse's human becoming theory of nursing. PMID- 10826307 TI - Political activity: a nursing intervention. PMID- 10826308 TI - Efavirenz. PMID- 10826309 TI - Secondary analysis of qualitative data: a means of collaboration in HIV-related research. PMID- 10826310 TI - International committee unveils initial plans to explore long-term use of HIV treatment. PMID- 10826311 TI - Endocrine therapy for breast cancer. AB - In addition to ovarian ablation by means of surgery or irradiation, a wide variety of endocrine agents are now available for the management of breast cancer, in both the metastatic and adjuvant settings. Currently available endocrine agents include antiestrogens, progestational drugs, and aromatase inhibitors. Randomized trials and meta-analyses have greatly clarified the specific roles of the more established of these agents in both premenopausal and postmenopausal women. However, the exact combination and sequence of these drugs are still under investigation. The addition of the bisphosphonates has also helped improve the quality of life of patients with metastatic disease. Among new endocrine agents currently under development are the pure antiestrogens, selective estrogen receptor modulators (SERMs), and a group of drugs with effects intermediate between the two. PMID- 10826312 TI - Adenocarcinoma of the esophagus: risk factors and prevention. AB - Esophageal cancer, with an estimated number of 12,300 new cases in the year 2000, is relatively uncommon in the United States but produces a high number of annual deaths, estimated at 12,100. Moreover, the incidence of the adenocarcinoma histologic type of esophageal cancer has been rising over the past two decades. Identification of risk factors could lead to primary prevention, as well as earlier diagnosis, treatment, and increased survival. Multiple risk factors are associated with the development of esophageal adenocarcinoma. These include Barrett's esophagus, acid peptic disorders, motor disorders of the esophagus, other malignancies, medications, environmental exposures, diet, and nutrition. However, no one particular risk factor is responsible for the rising incidence of esophageal cancer. Several preventive strategies are under investigation using such agents as nonsteroidal anti-inflammatory drugs (NSAIDs), selenium, alpha difluoromethylornithine (DFMO), and retinoids. As we gain more insight into the biology of this disease, other risk factors will hopefully be identified that will enable us to develop effective prevention strategies and, thus, reverse the current rising incidence of esophageal carcinoma. PMID- 10826313 TI - Cancer summit leads to global network to help unify research in fight against cancer. PMID- 10826314 TI - Biological basis of radiation sensitivity. Part 1: Factors governing radiation tolerance. AB - Local tumor recurrence after radiation therapy is due primarily to the failure to eradicate all of the tumor cells within the treatment fields. Theoretically, all cancers could be controlled locally if a sufficiently high radiation dose could be delivered to a treatment volume that encompassed all of the tumor cells. In practice, however, the administration of a radiation dose high enough to sterilize all of the tumor cells would pose an unacceptably high risk of severe damage to normal tissues. Technologic improvements in the delivery of therapeutic radiation have led to some improvements in the therapeutic ratio (i.e., the ratio of the dose required to eradicate every tumor cell to the dose that produces unacceptable normal tissue toxicity). Further significant improvements in the therapeutic ratio will drive from an understanding of the mechanisms governing the sensitivity of malignant and normal cells to radiation. Part 1 of this two part article reviews the clinical and tissue kinetic factors that govern the sensitivity of normal tissues and organs to ionizing radiation. Part 2, which will appear in next month's issue, describes recent insights into the cellular and molecular pathways that determine the sensitivity of normal cells and tumor cells to radiation. PMID- 10826315 TI - Clinical trials, referral resource. Treatment of advanced or recurrent squamous cell carcinoma of the head and neck. PMID- 10826316 TI - Management of patients with cancer of unknown primary site. AB - Cancer of unknown primary site represents approximately 3% to 5% of all new cancer diagnoses. Adenocarcinomas account for 60% of all unknown primary cancers and poorly differentiated carcinomas or adenocarcinomas, for 30%. Historically, the prognosis for most patients with unknown primary tumors has been poor, with survival often less than 6 months from diagnosis. Recent advances in diagnostic techniques, including immunocytochemical and molecular genetic methods, have increased the probability of identifying a likely underlying tumor type. Based on clinical and pathologic features, approximately 40% of patients can be categorized within subsets for which specific treatment has been defined. Empiric therapy is an option for the remaining 60% of patients. In these patients, favorable prognostic factors for treatment response include tumor location in lymph nodes, fewer sites of metastases, younger age, and poorly differentiated carcinoma histology. Although experience remains limited, the incorporation of a taxane into empiric regimens appears to improve response rates and survival. A recent study of paclitaxel (Taxol), carboplatin (Paraplatin), and etoposide in 55 patients with cancer of unknown primary site reported an overall response rate of 47% and a median overall survival of 13.4 months. Investigations continue to explore new diagnostic techniques and novel therapeutic approaches. PMID- 10826317 TI - Sedation in the imminently dying patient. AB - Sedation is a clinically important therapeutic intervention in the imminently dying patient. As the patient with an advanced, irreversible illness nears the end of life, symptoms accumulate that are progressively more difficult to manage and that may become refractory to standard medical interventions. The most common of these intractable symptoms are pain, agitated delirium, dyspnea, and existential or psychological distress. Various therapeutic options available for relieving these symptoms include physician-assisted suicide, euthanasia, acceptance of unrelieved suffering, and terminal sedation. Some commentators have voiced concerns that sedating the imminently dying patient inevitably hastens death, and that this practice, in fact, is a surrogate form of physician-assisted suicide or euthanasia. Ethical arguments invoked to support the use of terminal sedation include the principle of double effect, which draws a moral distinction between the intention of an act (in this case, to relieve suffering) and its foreseen but unintended consequence (premature death). This author views sedation as a necessary, although risk-laden, procedure that, if practiced by trained, dedicated clinicians, maintains the physician's twin obligations to benefit patients and to "do no harm." PMID- 10826318 TI - Smallpox vaccine. PMID- 10826319 TI - Assembly line medicine. PMID- 10826320 TI - Preschool vision screening: the importance of the two-line difference. PMID- 10826321 TI - Genetic screening and the pediatrician. PMID- 10826322 TI - The family history as a screening tool. PMID- 10826323 TI - Genetic testing in children: ethical and social points to consider. PMID- 10826324 TI - Advances in newborn screening for metabolic disorders: what the pediatrician needs to know. PMID- 10826325 TI - Universal screening for congenital hearing loss. PMID- 10826326 TI - [The attitude of pregnant women to the possibilities for the prenatal screening and diagnosis of Down's syndrome in the 2nd trimester]. AB - The aim of the study was to assess pregnant women's attitude and receptivity for second trimester prenatal screening and diagnostic tests for fetal Down syndrome [DS], factors that influence attitude formation, sufficiency of patients' information, advisability of introduction of these tests in routine prenatal care Interviews with 129 pregnant women were conducted after they had received written information concerning prenatal DS screening and diagnostic tests Five questions to the point of the matter as well as 14 related to the personal characteristics of the interviewed were included. Ultrasound screening rt]. accepted by 98.4% serum screening--by 93% and invasive prenatal testing--by 90% of the pregnant women Patients receptivity for serum screening and invasive testing Was influenced by factors like age, past obstetric history educational level, religiosity, attitude towards patient's own health For some of the factors statistically significant relationships were present while for others only some trends were outlined Regarding patients' high receptivity for prenatal DS screening and diagnostic tests the latter can be recommended as an element of the routine prenatal care in our country. PMID- 10826327 TI - [The Joel-Cohen (Misgav Ladach) method--a new surgical technic for cesarean section and gynecological laparotomy]. AB - The authors make a review on this problem for first time in Bulgarian literature and for the first time by us operated with the Misgav Ladach method--cesarean section. The authors describe 50 cases wit Ladach method and as control 20 with Pfannenstiel. The authors establish shorter duration of the operation-Si] and 56.3 min. respective in benefit for Misgav method; the babies were extracted at 5.5 and 12.3 min. respect. The mothers with Misgav had better prognosis at the time of dehospitalization. PMID- 10826328 TI - [The anthropometric indices, morbidity and mortality of newborn infants with diabetic fetopathy]. AB - A study was carried out to examine the anthropometric characteristics, neonatal morbidity and mortality in newborn infants with diabetic fetopathy. MATERIAL AND METHODS: Infant anthropometric measurements were obtained, neonatal morbidity and early neonatal mortality were determined in 15 neonates with diabetic fetopathy over 6 years. RESULTS: Among the study population, 33.3% of children had macrosomia. The incidence of aspiration syndrome and meconium aspiration syndrome was 40%, transient tachypnea was 33% and the observed incidence of hyaline membrane disease was 26%. The cardiologic pathology was result of patent ductus arteriosus, myocardial hypertrophy and persistent pulmonary hypertension. The most frequent biochemical changes were transient neonatal hypoglycemia and hyperbilirubinemia. The perinatal mortality rate was 26.7%, with cases of hemorrhagic disease of newborn and the incidence of neonatal thromboembolism. CONCLUSION: Despite the current improvements of diabetes care in pregnancy, newborn infants with diabetic fetopathy have a high level of morbidity and early neonatal mortality. PMID- 10826329 TI - [Asymptomatic infections of the genital tract and infertility]. AB - Diagnosis of male genital tract inflammations plays a significant role in andrology. Although genital infections are often silent they can severely impair male infertility. In seminal plasma of 100 patients were determined IgG and SigA (radial immunodiffusion) the number of peroxydase-positive cells in addition to conventional semen parameters and microbiological investigations. There was carried out quantitative determination of alpha-amylase in connection with semen liquefaction. The results showed that exact quantitation of above mentioned parameters distinguish inflammatory male adnexal affections, which is appropriate for control of anti-inflammatory treatment and facilitates the diagnosis of inflammatory process in andrology. PMID- 10826330 TI - [A comparative study between hysterosalpingography and laparoscopy in evaluating female infertility]. AB - The diagnostic accuracy of hysterosalpingography and laparoscopy was compared. One hundred fourty three infertile women were prospectively investigated in two years duration. Laparoscopic chromopertubation using methylen blue dye performed on days 8-16. Hysterosalpingography performed on days 7-10 with water soluble contrast medium. There was agreement between the results of both methods in 66.4 per cent /95/ of the patients. In 15 of the 63 patients with a normal hysterosalpingography, however, peritubal or tubal disease was observed at the time of laparoscopy/false-negative results, 10.4%/Eighty patients with tubal obstructions were detected by hysterosalpingography, but 33 obstructions were not confirmed by laparoscopy/false-positive results, 23%/. The greatest discrepancy between the hysterosalpingography and laparoscopy was found in peritubal adhesions--58.7%. Laparoscopic evaluation demonstrated also endometriosis in 41 cases, poly cystic ovaries in 12, ovarian cyst in 9, and uterine fibroids in 15. Intrauterine abnormalities were diagnosed by hysterosalpingography in only 2.9% per cent of the patients. No significant laparoscopic complications were noted. The results suggest that hysterosalpingography is useful as a primary screening procedure, but laparoscopy provides a more accurate assessment of tubal patency and peritoneal factors in the investigation of infertility. PMID- 10826331 TI - [The results of using ICSI at the Center for Assisted Reproduction, Varna]. AB - The authors present the results of 31 consecutive cycles of ICSI. Fertilization occurred in 251 /88%/ of 279 oocytes which resulted in 8 clinical pregnancies /27% per transfer/--3 singleton, 2 multiple uterine pregnancies, 2 abortions and 1 ectopic pregnancy. PMID- 10826332 TI - [The possibility for treating the premenstrual syndrome with Periodin]. AB - We examined the possibility of treatment of premenstrual syndrome (PMS) with Periodin (Walmark). Periodin is a combination of polyvitamins, oligoelements and phytoextracts which have a relation with prostaglandin's synthesis and regulation of prolactin secretion. A group of 24 women with PMS and regular cycles in premenopausal period (mean age of 44.7) was examined and took Periodin 1-2 caps daily for a period of 3 months as a food supplement. The assessment was made by modified J. Mortola score system. After treatment the PMS symptoms decreased with 52.3% compared with the beginning. The authors conclude that Periodin has a considerable influence on PMS symptoms, no contraindications and could be useful widely in gynecological practice. PMID- 10826333 TI - [The treatment of cyclic mastalgia--a comparative study between the preparations Bromocriptine and Geritamin]. AB - The aim of the prospective randomized study was to compare the reduction in cyclical breast pain after treatment with oral bromocriptine for 30 days (42 cases) and Geritamine for 40 days (44 cases). The overall response rate to first group was 78.5% and 64.0% to second group. Nine patients treated with bromocriptine and 15 cases with Geritamine have refractory cyclical mastalgia (not influence). The relapse rate after stopping bromocriptine was 30.3% and was 44.8%, with a mean follow-up of 5 months. The investigators indicate that these agents will be used for a twice per year. PMID- 10826334 TI - [Laparoscopic hysterectomy--the initial experience in Bulgaria]. AB - The paper presents the first cases of laparoscopic-assisted vaginal hysterectomy and laparoscopic hysterectomy with removal of the uterus through the vagina performed in Bulgaria as well as the experience gained by the team from II gynaecology clinic at State University Hospital "Maichin dom" in Sofia from seven patients operated on by these techniques. The indications, the conditions and the implemented technique with its advantages and difficulties are described. The procedures are associated with less trauma and blood loss than abdominal surgery and are followed by a very smooth postoperative period which is very well tolerated by the patients. No intraoperative complications occurred. The postoperative period was free of any problems in 5 of the patients while in two there were minor complications. Marked shortening of the duration of the procedures is achieved with the initial experience. PMID- 10826335 TI - [Disorders in sexual function after hysterectomy]. AB - The authors studied 120 women with hysterectomy, divided into 4 groups according to the volume of the surgical intervention (with preserved or removed cervix uteri and with preserved or eliminated ovaria). In the groups do not enter hysterectomies of oncological diseases and of patients with vaginal hysterectomy. The authors found a disturbed sexual function. Towards the different biological problems exist and a psychological one, the stress and the inhibited self-esteem from the loss of the important for the woman's functioning organs. In a small part of the operated patients was observed an increased sexual activity, as a psychological compensation of the inferiority feeling. PMID- 10826336 TI - [Transvaginal sonography, dilatation and curettage, computed tomography and hysteroscopy as methods for the diagnosis of the involvement of the cervical canal in cases of endometrial carcinoma]. AB - The quality of the preoperative diagnostic procedures D&C, hysteroscopy, transvaginal sonography, and computer tomography were evaluated. The results of these diagnostic methods were compared with the postoperative pathological results. The computer tomography was with limited abilities for determination of cervical canal involvement. The high negative predictive value of the rest of the methods gives us the ability to exclude cervical canal involvement with great accuracy and probability. But none of these methods could not on firm the presence of cervical canal infiltration, because of low percentage of positive predictive value, which was between 7.8% in hysteroscopy to 50% in transvaginal sonography and till 60% in MRI imaging. Between all these methods for diagnosis of cervical canal involvement--the transvaginal sonography was relatively efficient/78.2%/, specific in/79%/ and sensitive in /76%/ and relatively cheap. Because of this reason this method of transvaginal sonography is preferable in the evaluation of possible cervical infiltration. PMID- 10826337 TI - [Our experience in treating ovarian tumors with borderline malignancy]. AB - We tried to evaluate the treatment of borderline ovarian tumours for the period of 1987 till 1999. We analyzed 62 cases as the age varied from 16 till 82 years of age, median 51.2. To most of them was performed surgical treatment, and to some of them immediately second look operation. In 38% of the women under 35 years of age there were, recurrences, as for the all group of border line tumours it was 19% recurrences. The recurrences appeared lately usually after the fifth year. We examined the time of operation, the stage, the histological type, the number of pregnancies, number of recurrences, and the disease free interval. The results showed that the conservative surgery in young women leads to high niveau of pregnancies, and also high level of recurrences. The young patients with unilateral disease stage IA were treated with conservative surgery, with intraoperative staging, without adjuvant chemotherapy. The menopausal women were treated as malignant tumours/Total hysterectomy bilateral salpingo-oophorectomy, omentectomy and appendectomy/ We show the need of active follow up of the treated women especially the young ones who are treated conservatively. PMID- 10826338 TI - [Reconstructive surgery after mastectomy--the anatomical prerequisites]. AB - Reconstructive surgery of the breast after mastectomy occupies an important place in the complex treatment of breast cancer. The aim of the present study is to determine the capacity of the dermo-muscular flaps and to study the anatomical premises for their use under breast reconstructions. On the base of putrid material are created operative models of reconstructions of the breast with myocutaneous flaps m. rectus abdominis, m. latissimus dorsi and m. gluteus maximus. A study of the anatomical parameters of 64 female individuals, at an age from 51 to 73 years, has been conducted. It was found out that the myocutaneous flap m. rectus abdominis has an average volume of 590 cc and is formed basically of dermo-subdermal tissue (more than 80%). The myocutaneous flap m. latissimus dorsi has an average volume of 290 cc and is formed basically of muscular tissue (85%). The myocutaneous flap m. gluteus maximus has an average volume of 650 cc and is formed of subdermal and muscular tissue in the correlation 65/35%. The most suitable for breast reconstructions turns out to be the myocutaneous flap from m. rectus abdominis, since it provides sufficient amount of new tissue and uncomplicated surgical technique. PMID- 10826339 TI - [Excessive hydramnion--its management and complications during delivery]. PMID- 10826340 TI - [Anal incontinence following vaginal delivery]. PMID- 10826341 TI - [Endometriosis in the surgical scar from cesarean section]. AB - The frequency of endometriosis in and around the surgical scar from cesarean section is from 0.03%-0.04% to 0.8% in some reports. It is difficult to be diagnosed, in spite of the typical symptoms: aching swelling in the area of the surgical scar, which is influenced by the phases of menstruation. Sonography and fine-needle biopsy can be used, but it is usually diagnosed through the surgical treatment. Hystological diagnosis can also be a matter of some difficulty, as sometimes it is necessary to differentiate from adenocarcinoma, pseudomyxoma peritonei and metastatic carcinoma. The treatment is surgical. It is examined a case of endometriosis, which occurred in the surgical scar from a cesarean section about a year after the operation. The treatment is surgical by means of wide cutting-out of damaged musculus rectus abdominis and the surrounding tissues. After the operation a seroma is formed, which is healed with aspiration drainage and compressive bandage. Three years after operation a recurrence is not established. PMID- 10826342 TI - [Morphological diagnosis in gynecology: the joint activities of clinicians and pathologists]. PMID- 10826343 TI - [Abortion on demand. Its incidence and the reasons. Pre- and postabortion contraceptive consultation]. PMID- 10826344 TI - [Current management in pregnancy and epilepsy]. AB - Modern treatments make possible uneventful pregnancy and successful delivery for most of the epileptic women. Major risks however remain the teratogenic effects of some antiepileptic drugs and the obstetric complications they may cause. This implies careful planning of pregnancy for women with epilepsy and alertness for possible unwanted or undiagnosed (in the early stages) pregnancy in epileptic women during their reproductive years. The authors summarize current opinions in drug selections, specific treatment regimens and the obstetrics problems during pregnancy and delivery in epileptic women. PMID- 10826345 TI - [Pregnancy following testicular extraction of spermatozoa (TESE) and intracytoplasmic sperm injection (ICSI) in azoospermia]. PMID- 10826346 TI - [New methodological guidelines for the prevention and treatment of venous thromboses during pregnancy and in the puerperal period in Kuwait. Thrombophilia Clinic, Al-Adan Hospital]. AB - The high incidence of venous thromboembolic problems among women during pregnancy and puerperium in Kuwait require a new guidelines to cope with the increased number of thrombotic events and complications in this group. Pregnant ladies should be refer red in very early trimester to a Thrombophilia Clinic where they will be checked routinely for general and special laboratory tests accordingly. They will be classified to high risk category or low risk category. The high risk category group should receive a prophylactic measures throughout pregnancy and puerperium, while the low risk group do not need further action. Those who develop venous thrombosis from any categories should be classified in the high risk category during next pregnancies. PMID- 10826347 TI - Point-of-care HIV testing using simple/rapid HIV test kits: guidance for health care professionals. PMID- 10826348 TI - An integrated approach to communicable disease surveillance. PMID- 10826349 TI - Infectious diseases in England and Wales: July to September 1999. PMID- 10826350 TI - [More stringent rules concerning xenotransplantation proposed in a report: the risk of severe infection can not be excluded]. PMID- 10826351 TI - [Patients' attitudes to xenotransplantation: "I'll do anything, I don't want to die!"]. AB - Transplantation of neural tissue is an effective therapeutical approach in Parkinson's disease, but the method is constrained by the lack of suitable donor material. Embryonic neural tissue from pigs, xenografts, is considered as an alternative source of donor tissue. The attitudes towards neural tissue grafting in general and xenografts in particular were investigated by interviewing a group of patients with Parkinson's disease. The analysis revealed an ambivalence regarding xenografts. A pragmatic view, with priority placed on survival over ethical and other reservations, became apparent. PMID- 10826352 TI - [Rapid increase of the use of PET in cancer investigation--but Sweden lags behind]. AB - Glucose consumption in tissue can be measured using positron emission tomography (PET) and 18F-deoxyglucose (18FDG). Malignant tumors rely largely on anaerobic glycolysis and show very rapid glucose consumption, and can therefore be imaged using PET and 18FDG. PET has been shown to be useful in the evaluation of patients with e.g. lung cancer, colo-rectal cancer, malignant melanoma and malignant lymphoma, in terms of both diagnostic accuracy and cost-effectiveness. The clinical use of PET for workup of cancer patients is increasing rapidly in North America as well as in the European Union, but Sweden is lagging behind. PMID- 10826353 TI - [Increased incidence of adenocarcinoma of the esophagus and cardia. Reflux and obesity are strong and independent risk factors according to the SECC study]. AB - In a Swedish nationwide case-control study, gastroesophageal reflux and obesity were identified as strong and independent risk factors for esophageal adenocarcinoma. A moderately strong association was found with adenocarcinoma of the gastric cardia. No significant association was found with squamous cell carcinoma of the esophagus. With increasing duration and severity of reflux symptoms and with increasing body mass index (BMI) the risk increased in a dose dependent manner. When combined, reflux symptoms and obesity entailed greatly increased risk estimates, with relative risks exceeding 100 compared with persons with neither reflux symptoms nor obesity. However, because adenocarcinoma of the esophagus and gastric cardia are rarities, the absolute risk of developing these tumors was still not high. Our calculations revealed that even in the group with the highest risk, endoscopic surveillance is not readily recommended. Possible reasons for the increasing incidence of adenocarcinoma of the esophagus include 1) a suspected increase in the prevalence of reflux disease; 2) the increasing prevalence of obesity reported in western populations; and 3) the widespread use of medications that relax the lower esophageal sphincter and might cause reflux. All of these hypotheses suffer from inconsistencies that need to be solved before any firm conclusions can be drawn concerning the reasons for the increasing incidence of esophageal adenocarcinoma. PMID- 10826354 TI - [Attachment theory: a biological basis for the patient-physician relation?]. AB - Attachment theory is an evolutionary sociobiological theory of development which postulates that the propensity to make emotional bonds to a differentiated and preferred person, conceived as stronger and/or wiser, is a basic component of human nature, distinct from feeding and sexuality. Attachment behavior is present in germinal form in the neonate and continues to be present "from the cradle to the grave". When faced with loss, threat, distress or illness we tend to seek out an attachment figure from which we can obtain protection. Attachment theory has implications for the patient-doctor relationship. The doctor who is responsive to the patient's verbal and nonverbal attachment signals may be seen as a supplementary attachment figure. Doctors are powerful social figures, expected to manifest appropriate empathic behavior upon encountering the biologically grounded need for attention and listening. PMID- 10826355 TI - [Incidence of cancer after primary deep venous thrombosis or pulmonary embolism]. AB - We report a nationwide study based on a cohort of patients with a thromboembolic event (TE) defined as deep venous thrombosis and/or pulmonary embolism identified from the Danish National Registry of Patients from 1977 to 1993. Cancer occurrence in the cohort was determined by linkage to the Danish Cancer Registry. Expected number of cancer cases was estimated from age, sex and site-specific incidence rates. A total of 26,653 TE-cases were identified. We observed 1737 cases of cancer versus 1371 expected (SIR 1.3; 95% CI, 1.21 to 1.33). The risk was particularly high during the first months of follow-up, but declined rapidly hereafter to a constant level slightly above 1.0 one year after the TE. Forty percent of patients diagnosed with cancer within one year from the hospitalization for TE had distant metastases at the time of cancer diagnosis. The observed risk pattern provides evidence that most often a preclinical cancer gives rise to a subsequent TE. An aggressive search for hidden cancer in patients with TE is not warranted. PMID- 10826356 TI - [Do dare to perform myomectomy vaginally!]. PMID- 10826357 TI - [The parliamentary resolution on priority principles in health care. "The prioritization" groups are clinically applicable"]. PMID- 10826358 TI - [Unnecessary work of the research ethics committees? A lot of work is done with studies which are never to be published]. PMID- 10826359 TI - [Increased use of psychopharmaceuticals among young people--a problem not a cure!]. PMID- 10826360 TI - [Racing health insurance costs: reconstruct the rules and give incentives to all involved to reduce sick-leave]. PMID- 10826361 TI - [High cholesterol levels are discriminated by new therapeutic recommendations]. PMID- 10826362 TI - [Better therapeutic potential for azathioprine and mercaptopurine]. PMID- 10826363 TI - [The Ekaterinburg Medical Research Center of Prophylaxis and Health Care for industrial workers: history, present and prospects (70th anniversary of its foundation)]. PMID- 10826364 TI - [On hygienic standards of the allowable levels of suspended particles in the air]. AB - Elevation of average daily concentrations of suspended particles in air, even being within the MAC, could be associated with higher mortality in general population and deteriorated respiratory symptoms in children, especially in those having chronic respiratory diseases with bronchial obstruction. Discussion on reduction of MAC for suspended particles is expendient with wider practice of fraction measurement their concentrations. PMID- 10826365 TI - [The influence of "rigidity" and structure of fibrous dust on their biological activity]. AB - The authors represent experimental data on cytotoxic, fibrogenic and mutagenic effects of fibrous dusts--"soft" pulp fibers and "stiff" ones (chrysotile asbestos, carbon, basalt and fiber glass) in comparison with the nonfibrous analogs (antigorit, quartz DQ-12 and others). Viability of peritoneal macrophages was depressed more dramatically by "stiff" fibers vs. the "soft" ones. Mutagenic activity was associated with the "stiffness" degree of the dust particles. When compared to fibrous chemical dusts, nonfibrous ones appeared inert in micronuclear test. PMID- 10826366 TI - [Occupational medicine in aluminum and copper alloy production]. AB - Workers engaged into aluminum alloys production demonstrated high prevalence of occupational respiratory diseases caused by dust--pneumoconiosis and dust bronchitis, workers of copper alloys production were diagnosed as having early stage of chronic lead intoxication. PMID- 10826367 TI - [Clinical and x-ray diagnosis and diagnosis and classification of occupational fluorosis]. AB - The authors summarize longstanding experience in studies of chronic occupational intoxication caused by fluoric compounds. Findings are that diagnosis and clinical classification of occupational fluorosis should base on osteosclerotic changes in compact substance of peripheral bones, primarily of forearms and legs. The authors give clinical classification of occupational fluorosis. PMID- 10826368 TI - [Perinatal disorders of the central nervous system in children under ecologically unfavorable conditions]. AB - Regular medical and biologic etiologic risk factors of perinatal disorders of central nervous system in children give much less contribution into neurologic diseases development, than they give under ecologically unfavorable conditions when those factors induce most disorders. PMID- 10826369 TI - [Relationship between occupational respiratory pathology and cumulative exposure to dust]. AB - The authors concluded that "multiplicity of excess in MAC for predominantly fibrogenic dusts" value remains quite informative criterion in classification of work conditions and in forecasting the development of occupational respiratory diseases. Suggested in the new "Hygienic criteria for evaluation and classification of work conditions according to jeopardy and harm of occupational hazards, hardiness and stress of work" Manual, method determining harm degree of work conditions according to multiplicity of excess in reference dust loads is arduous but not so informative, and recommended values for reference dust loads are boosted and need correction. PMID- 10826370 TI - [Some results of the used methodology for the evaluation of risk in the Sverdlovsk region]. AB - The authors represent a scheme of conceptual model for methodology to evaluate health risk caused by environmental pollution. The model is based on analysis of literature and the authors' experience. PMID- 10826371 TI - [25-year experience of Sverdlovsk regional occupational center on the basis of the research institute]. AB - Sverdlovsk Regional Occupational Center based on the Research Institute has highly qualified staff, incorporates ambulatory department, hospital with modern diagnostic and therapeutic equipment. The Center performs multiple tasks, being an organizational and methodic, diagnostic and occupational examination institution using up-to-date advances in industrial medicine. PMID- 10826372 TI - [Drug-free management of homeostasis in patients with vibration disease]. AB - The authors present results of homeostasis studies in 239 patients having vibration disease due to local vibration and demonstrate drugs-free correction (physio- and balneotherapy) of disorders caused by vibration. High therapeutic efficiency meant improved peripheral hemodynamics of upper limbs, coagulation parameters, lipid metabolism and lipid peroxidation. PMID- 10826373 TI - [The use of atomic emission spectral method for metal detection in environment and biologic materials]. AB - Registering spectrum simultaneously by two devices enables to use atomic emission spectral method for various metals detection in environmental objects and biologic materials. PMID- 10826374 TI - [On the ration between solubility and cytotoxicity of silica modifications]. PMID- 10826376 TI - [Occupational evaluation of new reagent technologies for drinking water preparation]. PMID- 10826375 TI - [The evaluation of the efficiency of biological prophylaxis of lead poisoning (experimental study)]. PMID- 10826377 TI - [Status and prospects of using natural therapeutic resources in the Urals region]. PMID- 10826378 TI - The role of the sand in chemical warfare agent exposure among Persian Gulf War veterans: Al Eskan disease and "dirty dust". AB - The purpose of this paper is to inquire into the relationship between Al Eskan disease and the probable exposure to chemical warfare agents by Persian Gulf War veterans. Al Eskan disease, first reported in 1991, compromises the body's immunological defense and is a result of the pathogenic properties of the extremely fine, dusty sand located in the central and eastern region of the Arabian peninsula. The disease manifests with localized expression of multisystem disorder. Signs and symptoms of Al Eskan disease have been termed by the news media "Persian Gulf syndrome." The dust becomes a warfare agent when toxic chemicals are microimpregnated into inert particles. The "dirty dust" concept, that the toxicity of an agent could be enhanced by absorption into inactive particles, dates from World War I. A growing body of evidence shows that coalition forces have encountered Iraqi chemical warfare in the theater of operation/Persian Gulf War to a much greater extent than early U.S. Department of Defense information had indicated. Veterans of that war were exposed to chemical warfare agents in the form of direct (deliberate) attacks by chemical weapons, such as missiles and mines, and indirect (accidental) contamination from demolished munition production plants and storage areas, or otherwise. We conclude that the microimpregnated sand particles in the theater of operation/Persian Gulf War depleted the immune system and simultaneously acted as vehicles for low-intensity exposure to chemical warfare agents and had a modifying-intensifying effect on the toxicity of exposed individuals. We recommend recognition of a new term, "dirty sand," as a subcategory of dirty dust/dusty chemical warfare agents. Our ongoing research efforts to investigate the health impact of chemical warfare agent exposure among Persian Gulf War veterans suggest that Al Eskan disease is a plausible and preeminent explanation for the preponderance of Persian Gulf War illnesses. PMID- 10826379 TI - A descriptive analysis of patient encounter data from the Fleet Hospital FIVE humanitarian relief mission in Haiti. AB - Fleet Hospital FIVE personnel treated more than 24,000 patients during a 1997 military operation other than war in Haiti. Sample data were analyzed for 10,215 Haitians who received humanitarian assistance care at field treatment sites and for 353 military, United Nations personnel, and Haitian nationals who were treated at the fleet hospital. Demographic data, type of encounter, diagnoses, and prescriptions were tabulated. Children aged 1 to 10 years accounted for 31% of humanitarian assistance visits. Females outnumbered males; in adults aged 21 to 30 years, the proportion was almost three to one. Most (97%) were initial encounters. Infectious and parasitic diseases, such as worms or scabies, accounted for 25% of diagnoses. At the fleet hospital, more than 80% of patients were males; these were most often older than 21 years. Injuries and aftercare procedures constituted 51.5% of diagnoses. Of 18,100 prescriptions, 57% were for anti-inflammatories, vitamins, or anti-parasitics. Implications for medical planning are described. PMID- 10826380 TI - Maintenance of water and food quality in the stabilization forces area of operations: German surveillance laboratories and the problem of expired field rations. AB - The medical services of the German contingent of the Stabilization Forces in Bosnia maintain a high-standard-preventive health care system. Because knowledge about German food and water surveillance laboratories is not widespread, these and the methods used for sample examinations as well as their analytical capacities are introduced. From July to November 1998, we examined 168 food samples. Three percent of them were considered unsafe for consumption as a result of microbiological contamination. None was considered unsafe for chemical reasons. Consequently, only one case of suspected food-borne infection occurred. Similar results were found when we examined water quality. Generally, microbiological contamination was low. One sample of mineral water (0.5%) was considered unsafe for consumption for chemical reasons. These findings suggest a high-quality standard of food and water supply for the Stabilization Forces in Bosnia. The maintenance of this level is an important aim of the German preventive health care system. PMID- 10826381 TI - Medical ethics surveillance in the Armed Forces. AB - Modern defense services depend on a policy of the vigorous promotion of research to ensure that they retain an advantage in any future operational context. Research involving personnel within the armed forces, however, has certain constraints with respect to contemporary, best-practice medical ethics. Service members are one example of a class of "captive subjects" who require special protection in the context of medical research. (Prisoners, students, children, and the intellectually disabled are other such examples.) The majority of national defense forces now have ethical watchdog groups--institutional ethics committees--that oversee research involving service members. Such groups monitor the special considerations and constraints under which subjects in uniform can volunteer for biological research. These committees audit particularly the ethical themes of confidentiality, equality, and justice. Themes inherent in medical research in the military include the standard Beauchamp-Childress paradigm of autonomy, beneficence, nonmaleficence, and justice, to which are added the traditional military values of loyalty, respect, courtesy, and chivalry. Contemporary thinking is that the general principle of affording service members the opportunity to volunteer for research should be maintained within the constraints of compromised training time, national security, and operational necessity. Most biological research (and its outcome) does not in practice compromise confidentiality or military security. This paper presents an audit of the functioning of one national military medical ethics committee, the Australian Defence Medical Ethics Committee, and presents a discussion of its philosophies and influence within the broader military context. The Australian Defence Medical Ethics Committee believes that most research should, as an a priori condition of approval, be intended for open publication in peer-reviewed journals. PMID- 10826382 TI - The relationship between heavy alcohol use and work productivity loss in active duty military personnel: a secondary analysis of the 1995 Department of Defense Worldwide Survey. AB - This cross-sectional study examines the association between heavy alcohol use among active duty military personnel and five work productivity loss events that may have an adverse effect on military performance and readiness. Data for light (N = 3,147) and heavy (N = 2,242) drinkers, categorized by gender and pay grade, were obtained from the 1995 Department of Defense Worldwide Survey. Drinking classification was predefined using a standard algorithm that factored quantity and frequency of wine, beer, and liquor consumed. The relative risks of experiencing a productivity loss event at a particular level and 95% confidence intervals were calculated by applying the Mantel-Haenszel method after adjusting for age. The relative risks for increased self-reported lateness, leaving early, low performance, and on-the-job injury were all higher for heavy drinkers than for light drinkers. This association between the heavy-drinking population and four of five work productivity loss events indicates that prevention programs should target all personnel. PMID- 10826383 TI - Why is Marine combat mortality less than that of the Army? AB - Data from recent wars indicate that a wounded Marine had a 20% lower risk of dying than an Army soldier. Possible reasons for this difference are (1) Navy care is superior, (2) soldiers sustained more severe wounds, and (3) the services count casualties differently. Injury severity was measured in random samples of Marines and soldiers that were selected from the Wound Data and Munitions Effectiveness Team database. There was no difference in the lethality of injury and the prevalence of lifesaving first aid. Wounded Marines were more likely to wear protective vests, and this decreased Marine mortality in Vietnam. Hospitalized Marines had lower Injury Severity Scores and were less likely to be returned to duty without first being admitted to a medical treatment facility. Lower Marine combat mortality is primarily the result of the fact that a Marine with a minor soft tissue wound was more likely to be hospitalized than was a soldier with a similar injury. PMID- 10826384 TI - The incidence of known diabetics among U.S. Army active duty dental patients. AB - Diabetes affects an estimated 16 million Americans. The purpose of this study was to determine the incidence of known diabetes among active duty soldiers by reviewing medications dispensed for diabetes at a large military installation. This study was conducted from July 31, 1997, through August 1, 1998. Fifty-eight (0.18%) members of the total active duty male population of 32,593 had been dispensed diabetes medications, whereas 43 (0.48%) members of the total female active duty population of 8,898 had been dispensed medications for diabetes. Gestational diabetes was the cause for 88.37% of the total active duty females dispensed diabetic medication at Fort Hood, Texas. The female-to-male ratio for diabetes was 2.7:1. Diabetics made up 0.27% of the total population. PMID- 10826385 TI - Carpal tunnel syndrome: effect on Army dental personnel. AB - The purpose of this study was 2-fold: (1) to determine the prevalence of hand problems, in particular carpal tunnel syndrome (CTS), among Army dental personnel; and (2) to identify dental professionals at risk. A 12-page survey was mailed to all U.S. Army military and civilian dental personnel. Of the 6,320 surveys mailed, 80.9% were returned completed. An analysis was performed identifying the prevalence of hand problems and CTS and noting differences between civilian and military dental personnel. Of the 5,115 surveys analyzed, 44.8% indicated hand problems and 25.4% were determined to indicate a high probability of CTS. Of the 18 dental job specialties, dental therapy assistants and dental hygienists had the highest prevalence of CTS, 73% and 57%, respectively. Army dental personnel are at greater risk of developing CTS than the general public, especially civilian dental personnel, who were female, older, and employed longer. PMID- 10826386 TI - Evaluation of possible battlefield tourniquet systems for the far-forward setting. AB - A significant number of casualties in previous conflicts died from peripheral vascular wounds. A well-designed tourniquet could possibly have prevented these deaths. The objective of this study was the identification of such a tourniquet. A survey of Special Operations corpsmen established important characteristics necessary in an ideal tourniquet. Because most available devices do not and patented ideas could not meet these criteria, a number of prototypes were developed. Seven potentially satisfactory tourniquets were evaluated by 15 Navy SEAL corpsmen. The success and timing of placement were recorded, and a follow-up questionnaire was completed. Of the several successful tourniquets, two were preferred. Tourniquets incorporating a windlass technique take longer to place and often fail when placed with only one hand. New, relatively simple tourniquet devices incorporating bladder and ratchet mechanisms can significantly improve tourniquet performance. PMID- 10826387 TI - Designing a carotid endarterectomy critical pathway for your organization. AB - BACKGROUND: Carotid endarterectomy (CEA) is one of the top-five surgical diagnosis-related groups at Keesler Medical Center. The geometric mean length of stay for CEA during fiscal year (FY) 1996 was 5.84 days (N = 41), compared with 1.79 for a benchmark facility. OBJECTIVE: Create a critical pathway to standardize care, maintain/improve patient outcomes, reduce lengths of stay, and decrease costs. METHODS: A multidisciplinary team was formed to evaluate four patient-flow options. The team decided to discharge patients directly from the intensive care unit to meet both patient and staff needs. RESULTS: The geometric mean length of stay decreased to 1.70 days (N = 54) in FY 1998, compared with 2.42 days (N = 40) in FY 1997. The cost savings ws $5,841 per case, compared with $1,684 before creation of the pathway. This represents an annual savings of more than $224,000 and a 30% reduction in length of stay. CONCLUSIONS: The CEA pathway has standardized the care received by this group of patients. By decreasing variation, processes have become routine and more efficient. PMID- 10826388 TI - Primary care access improvement: an empowerment-interaction model. AB - Improving community primary care access is a difficult and dynamic undertaking. Realizing a need to improve appointment availability, a systematic approach based on measurement, empowerment, and interaction was developed. The model fostered exchange of information and problem solving between interdependent staff sections within a managed care system. Measuring appointments demanded but not available proved to be a credible customer-focused approach to benchmark against set goals. Changing the organizational culture to become more sensitive to changing beneficiary needs was a paramount consideration. Dependent-group t tests were performed to compare the pretreatment and posttreatment effect. The empowerment interaction model significantly improved the availability of routine and wellness type appointments. The availability of urgent appointments improved but not significantly; a better prospective model needs to be developed. In aggregate, appointments demanded but not available (empowerment-interaction model) were more than 10% before the treatment and less than 3% with the treatment. PMID- 10826389 TI - Factors affecting Department of Defense patient satisfaction in a military emergency department. AB - This descriptive, correlational study was conducted to explore factors (nurses and staff, doctors, waiting and convenience issues, and tests and treatments) and specific interpersonal behaviors affecting Department of Defense patient satisfaction in a military emergency department (ED). Data from a random sample of 534 patients using a mail-back questionnaire (N = 178) revealed that military patients who perceived the staff to have treated them courteously, taken their problems seriously, and paid attention to them during their stay were more satisfied with their military ED encounter. Patient satisfaction, expressed as likelihood to recommend the facility, was independent of major demographic characteristics such as age and sex. Our findings suggest that a patient's perception of the ED encounter, particularly the interactions between the patient and staff, ultimately influence satisfaction. Significant improvements in patient satisfaction might be realized if quality improvement efforts focus on providing ED staff specific training in interpersonal interactions. PMID- 10826391 TI - The use of deployable military hospitals after hurricanes: lessons from the Hurricane Marilyn response. AB - Deployable military hospitals are frequently requested to supplement surviving local health care capabilities after disasters. Although some authors have advocated their use after mass casualty events such as earthquakes, previous reports have questioned the appropriateness of deploying these hospitals after destructive storms. These hospitals are relatively slow moving, expensive, and may require the diversion of local resources to support. After Hurricane Marilyn in 1995, a military hospital was deployed to the U.S. Virgin Islands. For a variety of reasons, the local health care community declined to use the facility once it was operational. This report is based on interviews with disaster managers and local health officials involved in the Hurricane Marilyn response. Recommendations include improving communications while requesting resources, broadening the range of available health assets to increase flexibility, positioning resources regionally or in the civilian sector, and creating clear indications for full-scale deployable hospitals when they are required. PMID- 10826390 TI - Providing managed care options for a large population: evaluating the CHAMPUS reform initiative. Civilian Health and Medical Program of the Uniformed Services. AB - OBJECTIVES: To evaluate a managed care demonstration project in CHAMPUS (Civilian Health and Medical Program of the Uniformed Services), the insurance program covering physical and mental health care services for the dependents of active duty military personnel, military retirees, and the retirees' dependents. The demonstration project added a health maintenance organization (HMO) option and a preferred provider organization (PPO) option to the standard CHAMPUS coverage and allowed beneficiaries to select the coverage option they preferred. DATA SOURCES: Utilization, costs, access, and beneficiary satisfaction were measured using data from CHAMPUS claims records, the Defense Enrollment Eligibility Reporting System, the demonstration project contractor's HMO enrollment file, the contractor's list of network hospitals, and two surveys of CHAMPUS beneficiaries. STUDY DESIGN: Changes in utilization at 11 demonstration sites were compared with changes in utilization at 11 matched control sites. The effect of the demonstration project on costs was evaluated by estimating the costs for the demonstration sites both with and without the managed care options based on data from the control sites. Access to care and satisfaction were compared between the demonstration sites and control sites based on beneficiary surveys. DATA COLLECTION: All claims in both demonstration and control sites were used in estimating utilization changes. Two mailed surveys were sent to a randomly selected sample of active duty and retiree households with CHAMPUS beneficiaries; the sample was stratified by beneficiary type (active duty or retiree) and site. PRINCIPLE FINDINGS: Overall utilization in the CHAMPUS system decreased at the demonstration sites but stayed approximately the same at the control sites. Utilization among the enrollees in the HMO demonstration option, however, increased dramatically. Patient access to care and satisfaction generally remained at the same levels at both demonstration and control sites, but enrollees in the HMO option reported higher satisfaction. Costs to the government at the demonstration areas, based on regression estimates from the control sites, were about the same or slightly higher than what they would have been under the standard CHAMPUS system. CONCLUSIONS: Managed care plans for large government-sponsored insurance programs can reduce utilization and maintain patient access and satisfaction. Careful structuring of such plans is needed, however, if they are to reduce costs. PMID- 10826392 TI - The backpack run test: a model for a fair and occupationally relevant military fitness test. AB - Our purpose in this investigation was to develop and validate a theoretical model for a backpack run test based on how fast one can run 2 miles while wearing a backpack. Using actual unloaded (no backpack) 2-mile-run test data from 59 male service academy cadets, we calculated the average oxygen cost during the run, the equivalent cost if wearing additional weight, and the corresponding estimated run time with the backpack. The correlations between body weight and loaded (backpack weight = 30 kg) run times (r = 0.55 [p < 0.05] and r = 0.12 [p > 0.05], respectively) demonstrate that the bias against heavier runners is eliminated with the backpack run. Given that the backpack run test requires only standard issue equipment, demonstrates clear occupational and health-related fitness relevance, predicts no apparent body-size bias, and measures work- and health related components of fitness, we recommend that the military services consider the present data when developing or modifying tests of physical fitness. PMID- 10826393 TI - Premature treatment termination by angry patients with combat-related post traumatic stress disorder. AB - Angry patients with conjoined post-traumatic stress disorder often direct their anger at health care providers during the course of treatment. Such misplaced anger can interfere with treatment. Emerging treatments for trauma-related anger are effective. However, even in the course of psychotherapy for trauma-related anger, these patients direct anger at their therapists, compromising the treatment alliance and increasing the likelihood of premature termination. A case example is presented to illustrate the effect of anger on the treatment alliance. A therapeutic strategy is proposed to reduce the likelihood of premature treatment termination in these high-risk patients. This strategy may also be helpful in primary care contexts. PMID- 10826394 TI - Congenital radioulnar synostosis in an active duty soldier: case report and literature review. AB - Congenital radioulnar synostosis (CRS) is a developmental deformity that interferes with pronation and supination of the upper extremity. CRS often results in functional, cosmetic, and cultural limitations. The purpose of this paper is to present a patient with CRS who is currently functioning as a generator mechanic in the U.S. Army and to review the literature for current treatment options. We know of no other individual in the U.S. Army who is on active duty status with this deformity. PMID- 10826395 TI - Practitioner's Trial on the Efficacy of Antihypertensive Treatment in the Elderly Hypertension (The PATE-Hypertension Study) in Japan. AB - Patients aged 60 years and older with essential hypertension were treated with an angiotensin-converting enzyme inhibitor (ACE-I), delapril (Adecut) or a long acting calcium (Ca)-antagonist, manidipine (Calslot) for 3 years. The incidences of cardiovascular events as well as drug-related side effects were compared between the two groups to investigate whether both classes of antihypertensive drugs are beneficial in elderly hypertensive patients. There were no significant differences in characteristics of patients between the two intervention groups, except for slightly lower blood pressure (P = .08) in the Ca-antagonist group at the initiation of the study. There were no significant differences in total death between the two groups. Cardiovascular events (both fatal and nonfatal) were noted in 34 of 699 patients (22.5/1000 patient-years) in the ACE-I group and 50 of 1049 patients (19.7/1000 patient-years) in the Ca-antagonist group, with no significant difference found between the two groups. The correlation between cardiovascular incidence and the blood pressure attained during treatment showed a J-shaped phenomenon and suggests that an excessive reduction less than 120 mm Hg in systolic blood pressure (SBP) is unnecessary and may be harmful in certain cases. Side effects were more frequent in the ACE-I group than in the Ca antagonist group (P = .01). Cough was the major adverse event, occurring in 5.0% of patients in the ACE-I group. In conclusion, the study indicates that both ACE I (delapril) and Ca-antagonist (manidipine) were equally beneficial for reducing cardiovascular morbidity and mortality in elderly hypertensive patients. However, tolerability of ACE-I was lower due to the adverse event of coughing. PMID- 10826396 TI - The addition of doxazosin to the therapeutic regimen of hypertensive patients inadequately controlled with other antihypertensive medications: a randomized, placebo-controlled study. AB - This randomized, double-blind, placebo-controlled study evaluated the use of doxazosin as an add-on therapy for inadequately controlled hypertension. Patients with a sitting diastolic blood pressure (BP) of 95 to 115 mm Hg received either doxazosin (n = 38) or placebo (n = 32) in addition to one or two baseline antihypertensive medications. After an upward titration period, patients were maintained on a fixed dosage of doxazosin (1 to 16 mg/day) or matching placebo for 4 weeks. Doxazosin add-on therapy led to improvements, compared with placebo, in sitting systolic BP (adjusted mean change = -20.9 v -8.5 mm Hg, P = .001), sitting diastolic BP (-13.0 v -8.1 mm Hg, P = .026), and standing systolic BP ( 22.0 v -11.5 mm Hg, P = .011). Baseline antihypertensive therapy was gradually tapered or discontinued in patients who achieved a target reduction in BP (sitting diastolic BP of < 90 mm Hg in addition to a minimum improvement of 10 mm Hg in sitting diastolic BP over baseline) with add-on therapy (55% [n = 21] with doxazosin, 31% [n = 10] with placebo). Twelve patients in the doxazosin group maintained the target reduction in BP after complete withdrawal of their baseline antihypertensive therapy, compared with none in the placebo group. A small but statistically significant positive effect on the lipid profile was seen in the doxazosin group during add-on therapy. Doxazosin treatment was well tolerated, with an adverse event profile similar to that of placebo. These findings demonstrate that doxazosin add-on therapy is an effective, well-tolerated treatment strategy for patients with inadequately controlled hypertension. PMID- 10826397 TI - Additive pressor effects of caffeine and stress in male medical students at risk for hypertension. AB - The effects of caffeine on blood pressure (BP) and cortisol secretion were examined during elevated work stress in medical students at high versus low risk for hypertension. Among 31 male medical students who were regular consumers of caffeine, 20 were considered at low risk for hypertension (negative parental history and all screening BP < 125/78 mm Hg) and 11 at high risk based on epidemiologic criteria (positive parental history and average screening BPs between 125/78 and 139/89 mm Hg). Cortisol levels and ambulatory BP were measured with and without caffeine during two lectures (low work stress) and two exams (high work stress) in a randomized, double-blind, crossover trial. Caffeine consumption and exam stress increased cortisol secretion in both groups (P < .05). BP increased with caffeine or exam stress in both groups, low versus high risk, respectively (Caffeine: + 5/4 vs + 3/3 mm Hg; Stress: + 4/1 vs + 7/3 mm Hg; P < .05). The combination of stress and caffeine caused additive increases in BP (Low Risk + 9/5 mm Hg, High Risk + 10/6 mm Hg) such that 46% of high-risk participants had average systolic BP > or = 140 mm Hg. This combined effect of stress and caffeine on BP suggests that it may be beneficial for individuals at high risk for hypertension to refrain from the use of caffeinated beverages, particularly at times when work demands and attendant stressors are high. For the same reasons, recent intake of caffeine should be controlled in patients undergoing BP measurement for the diagnosis of hypertension. PMID- 10826398 TI - Association between alcohol intake and development of hypertension in Japanese normotensive men: 12-year follow-up study. AB - Although it has been suggested that alcohol intake is related to hypertension, few long-term prospective studies have investigated this relationship. We therefore conducted a prospective study in male residents of a farming community in Japan to determine whether heavy drinking would predispose to the development of hypertension. A total of 325 normotensive (< 140/90 mm Hg) men were enrolled in 1977. Twelve years later, 93 (28.6%) subjects became hypertensive (defined as blood pressure > or = 140/90 mm Hg or use of antihypertensive medication). The probability of the development of hypertension in heavy drinkers predicted from a logistic regression equation was 44.6% (relative risk: 2.05 versus nondrinkers) after adjusting for age and body mass index (BMI). It was 36.2% (relative risk: 1.86 versus nondrinkers) after a further adjustment for systolic blood pressure at baseline. A high odds ratio of 2.39 for the development of hypertension with alcohol intake of < 46 g/day versus > or = 46 g/day at baseline was obtained even after adjustments for age, BMI, and confounding factors. We conclude that habitual heavy drinking of alcohol is a risk factor for the development of hypertension. This is the first report demonstrating a significant relationship between habitual alcohol intake and the development of hypertension in a long term prospective study in Japan. PMID- 10826399 TI - Complex segregation analysis of blood pressure and heart rate measured before and after a 20-week endurance exercise training program: the HERITAGE Family Study. AB - Complex segregation analysis of baseline resting blood pressure (BP) and heart rate (HR) and their responses to training (post-training minus baseline) were performed in a sample of 482 individuals from 99 white families who participated in the HERITAGE Family Study. Resting BP and HR were measured at baseline and after a 20-week training program. Baseline resting BP and HR were age-adjusted and age-BMI-adjusted, and the responses to training were age-adjusted and age baseline-adjusted, within four gender-by-generation groups. This study also analyzed the responses to training in two subsets of families: (1) the so-called "high" subsample, 45 families (216 individuals) with at least one member whose baseline resting BP is in the high end of the normal BP range (the upper 95th percentile: systolic BP [SBP] > or = 135 or diastolic BP [DBP] > or = 80 mm Hg); and (2) the so-called "nonhigh" subsample, the 54 remaining families (266 individuals). Baseline resting SBP was influenced by a multifactorial component (23%), which was independent of body mass index (BMI). Baseline resting DBP was influenced by a putative recessive locus, which accounted for 31% of the variance. In addition to the major gene effect, which may impact BMI as well, baseline resting DBP was also influenced by a multifactorial component (29%). Baseline resting HR was influenced by a putative dominant locus independent of BMI, which accounted for 31% of the variance. For the responses to training, no familiality was found in the whole sample or in the nonhigh subsample. However, in the high subsample, resting SBP response to training was influenced by a putative recessive locus, which accounted for 44% of the variance. No familiality was found for resting DBP response to training. Resting HR response to training was influenced by a major effect (accounting for 35% of the variance), with an ambiguous transmission from parents to offspring. PMID- 10826400 TI - Effect of ACE inhibition on pressor, renal vascular, and adrenal responses to infusion of angiotensin I in normal subjects eating a low-salt diet. AB - To examine the influence of angiotensin-converting enzyme (ACE) on pressor, renal vascular, and adrenal responses during angiotensin I (Ang I) infusion, we studied 10 normotensive, healthy men. Each was in balance with a 10-mEq sodium, 100-mEq potassium intake and was studied before and during ACE inhibition with enalapril. Ang I (3, 10, and 30 ng/kg/min) was infused in each subject. Then ACE inhibition was instituted with enalapril for 3 days, which induced the anticipated fall in blood pressure, plasma Ang II, and aldosterone concentration, and rise in renal plasma flow. During ACE inhibition only the 30-ng/kg/min Ang I dose raised plasma Ang II levels. There was a spectrum, however, in the end-organ response to Ang I during ACE inhibition. Responses of plasma aldosterone concentration and blood pressure were in excellent accord with the reduction in Ang II formation. On the other hand, responses of the renal blood supply were substantially less inhibited than anticipated. Under the conditions of this study, ACE inhibition led to nonuniform changes in the response to exogenous Ang I, suggesting intrarenal conversion of Ang I to Ang II. PMID- 10826401 TI - Angiotensin II type 1 (AT1) receptor blockade in hypertensive women: benefits of candesartan cilexetil versus enalapril or hydrochlorothiazide. AB - The aim of this large, randomized, double-blind, parallel-group study in hypertensive women was to compare the antihypertensive efficacy and effects on subjective symptoms and quality of life of the new angiotensin II type 1 (AT1) receptor blocker candesartan cilexetil, the angiotensin-converting enzyme inhibitor enalapril, and the diuretic hydrochlorothiazide (HCTZ). Women, aged 40 to 69 years, with a seated diastolic blood pressure (DBP) of 95 to 115 mm Hg, were randomized to candesartan cilexetil, 8 to 16 mg (n = 140), enalapril, 10 to 20 mg (n = 146), or HCTZ, 12.5 to 25 mg (n = 143), for 12 weeks; the higher doses were used if DBP was greater than 90 mm Hg after 6 weeks. Candesartan cilexetil lowered seated blood pressure by 17/11 and 19/11 mm Hg after 6 and 12 weeks of treatment, respectively. This reduction was greater (P < .01) than with enalapril (12/8 and 13/9 mm Hg) or HCTZ (12/7 and 13/8 mm Hg). The proportions of patients with controlled DBP (< 90 mm Hg) after 12 weeks of treatment with candesartan cilexetil, enalapril, or HCTZ were 60%, 51%, and 43%, respectively. Patients experienced less dry cough (P < 0.001) with candesartan cilexetil or HCTZ than with enalapril. No treatment differences were found in the incidence of dizziness and quality of life was well maintained in all groups. Compared with candesartan cilexetil and enalapril, HCTZ increased uric acid and decreased serum potassium (P < .001). In conclusion, candesartan cilexetil reduced blood pressure more effectively and was better tolerated than enalapril or HCTZ in women with mild to moderate hypertension. PMID- 10826402 TI - The antiarrhythmic potential of angiotensin II antagonism: experience with losartan. AB - A large body of literature accumulated over the past several years supports the notion that inhibition of the renin-angiotensin system protects the heart and other target organs from hypertensive complications. Various studies have shown that angiotensin-converting enzyme inhibitors reduce morbidity and mortality in the setting of ischemic heart disease and/or congestive heart failure. The improvement in survival has been attributed in part to a significant decrease in the incidence of sudden deaths, possibly due to a decrease in complex arrhythmia episodes. Recently, the angiotensin II type 1 receptor antagonist losartan was shown to reduce mortality by 46% compared with captopril in older patients with chronic congestive heart failure. This paper briefly reviews the arrhythmogenic properties of angiotensin II and the possible pharmacologic mechanisms for the antiarrhythmogenic potential of losartan. PMID- 10826403 TI - Effect of amlodipine on hemodynamic and endocrine responses to mental stress. AB - Little is known about the effects of antihypertensive drugs on hemodynamic responses to mental stress. We studied 24 patients with mild-to-moderate hypertension in a double-blind random-sequence crossover study comparing placebo with amlodipine titrated up from 5 to 10 mg daily. After 1 month of treatment, the subjects performed 20 min of a frustrating cognitive task. At baseline before task, amlodipine significantly reduced systolic pressure (128.9+/-8.2 mm Hg v 140.3+/-10.7 mm Hg, P < .001), diastolic pressure (81.7+/-7.7 mm Hg v 90+/-7.5 mm Hg, P < .001), and total peripheral resistance (37.5+/-15 v 45.6+/-23.7 mm Hg/L/min, P < .05), while elevating baseline norepinephrine levels (2286+/-731 pmol/L v 1788+/-546 pmol/L, P < .001). Blood pressure during the stress task was significantly less with amlodipine than with placebo (systolic 142.3+/-12.3 mm Hg v 150.9+/-14.6 mm Hg, P < .001; diastolic 87.9+/-8.4 mm Hg v 97.7+/-9.3 mm Hg, P < .001), whereas norepinephrine was significantly higher (2754+/-1007 pmol/L v 1970+/-740 pmol/L, P < .001). There were no significant differences in cardiac output, plasma lipids or lipoproteins, or markers of platelet activation. Heart rate increased significantly during stress, but there was no significant difference between amlodipine and placebo either at baseline or during stress. Our conclusion is that amlodipine reduces blood pressure at baseline and during mental stress, but raises basal and stress-related plasma catecholamines. This finding may have implications for the recent controversy over the safety of calcium channel antagonists, and suggests the potential relevance of combining amlodipine with adrenergic blockers. PMID- 10826404 TI - Clinical relevance of office underestimation of usual blood pressure in treated hypertension. AB - Average 24-h blood pressure (BP) is more representative of usual BP than office BP. However, the clinical relevance of 24-h BP in treated hypertensive subjects is incompletely known. Thus, we studied 395 uncomplicated hypertensive subjects (209 men, 53+/-10 years) who were receiving antihypertensive drug therapy from >1 year. All subjects underwent 24-h ambulatory BP monitoring and M-mode echocardiography. Subjects were classified by tertile of the difference between observed and predicted 24-h systolic BP (the latter determined by regressing 24-h systolic BP on office systolic BP): higher-than-predicted (III tertile), around the regression line (II tertile), and lower-that-predicted (I tertile) 24-h BP. Despite similar office BP (144/89, 141/88, and 144/89 mm Hg in the III, II, and I tertile, P = not significant), age, body mass index, and duration of hypertension, left ventricular mass was greater in the subjects with higher-than predicted 24-h systolic BP (50+/-14 g x m(-2.7)) than in the other two groups (46+/-13 g x m(-2.7) and 42+/-10 g x m(-2.7), both P < .05). The III tertile also showed a more concentric left ventricular geometric pattern (relative wall thickness was 0.42+/-0.08, 0.40+/-0.07, and 0.38+/-0.07 in the III, II, and I tertile, P < .001) and a reduced systolic function at the midwall level (16.8+/ 3, 17.7+/-3, and 18.2+/-3, P < .001). In conclusion, treated hypertensive subjects whose 24-h BP is notably higher than one would predict from office BP are more likely to develop left ventricular hypertrophy, a strong adverse prognostic marker. In a sizable subset of treated hypertensive subjects, BP measured in the physician's office underestimates usual BP and its impact on left ventricular structure. PMID- 10826405 TI - Hypertension in cafeteria-fed rats: alterations in renal alpha2-adrenoceptor subtypes. AB - Obesity is a major cause of human essential hypertension and there are clear evidences that abnormal kidney functions play a key role in obesity hypertension. Feeding rats a cafeteria diet has been extensively used as an experimental model to study obesity and energy balance expenditure. The present study investigated whether rats fed a cafeteria diet develop hypertension with alterations in renal alpha2-adrenoceptor subtype distribution. Weight gain induced by feeding rats a cafeteria diet during 8 weeks was associated with a marked increase in blood pressure. Insulin levels were higher in these hypertensive rats, leading to a decreased plasma glucose/insulin ratio. Based on radioligand-binding studies using [3H]-RX821002 and selective competitors, a raise in alpha2-adrenoceptor density that was solely due to an increased alpha2B-adrenoceptor subtype density was detected in the kidney of the cafeteria-fed rat. Furthermore, reverse transcription-polymerase chain reaction (RT-PCR) experiments showed an overexpression of the gene encoding the alpha2B-adrenoceptor subtype in these rats. On the other hand, despite a similar mRNA level, the alpha2A-adrenoceptor subtype was no more detectable by radioligand-binding studies in the kidney of the cafeteria-fed rat. In conclusion, cafeteria-fed rats are hypertensive, with renal alterations in alpha2-adrenoceptor distribution. These alterations, which are not related to genetic factors, may play a key role in the onset of hypertension. PMID- 10826406 TI - Effect of protease inhibitors on angiotensin-converting enzyme activity in human T-lymphocytes. AB - The purpose of these investigations was to determine whether the aminopeptidase B and leucine aminopeptidase inhibitor bestatin, the chymase inhibitor chymostatin, the calpain inhibitor E-64, and the neutral serine protease inhibitor leupeptin affect the angiotensin converting enzyme (ACE) activity in T-lymphocytes. ACE activity in homogenates of T-lymphocytes or in intact T-lymphocytes in suspension was measured by determining fluorimetrically histidyl-leucine, formed from the conversion of hippuryl-histidyl-leucine, coupled with ophtaldialdehyde. The effect of various concentrations (10(-9) to 10(-3) mol/L) of the angiotensin converting enzyme inhibitors lisinopril and captopril and of the various protease inhibitors on ACE activity was studied. Lisinopril and captopril reduced the ACE activity in homogenates of T-lymphocytes in a concentration-dependent manner. Lisinopril exhibited a more pronounced inhibition of ACE in T-lymphocytes than did captopril. Chymostatin and E-64 had no effect on the ACE activity in T lymphocytes, whereas leupeptin inhibited its activity in a dose-dependent fashion. Bestatin, on the contrary, increased the ACE activity in homogenates of T-lymphocytes as well as in intact T-lymphocytes in proportion to the concentration. Our data showed that the ACE activity in T-lymphocytes was stimulated by bestatin and inhibited by leupeptin, whereas chymostatin and E-64 did not affect the ACE activity in T-lymphocytes. PMID- 10826407 TI - NaCl-induced hypertensive rat model of non-insulin-dependent diabetes: role of sympathetic modulation. AB - Systemic hypertension is common in individuals with non-insulin-dependent diabetes (NIDD) and, in this population, markedly increases the risk for cardiovascular complications. The aims of this study were to develop a rat model of combined NaCl-induced hypertension and NIDD, and to determine the contribution of the sympathetic nervous system to the development of the manifest hypertension. Two-day old male Wistar-Kyoto rats were injected with either streptozotocin (90 mg/kg, ip; NIDD) or vehicle (citrate buffer; control). At 4 weeks of age, the animals underwent either a right nephrectomy or a sham operation. Animals in each group were further subdivided, with one group maintained on normal (0.72 %) NaCl diet whereas the other was placed on a high (8%)-NaCl diet. At 6 months of age, diabetes was confirmed by glucose tolerance testing. Hemodynamic parameters were measured in the freely moving animal (ia) before and after the administration of prazosin (peripheral alpha1-adrenergic antagonist, iv) or clonidine (central alpha2-adrenergic agonist). The NIDD rat displayed a higher (P < .05) blood glucose concentration than the nondiabetic control rat during the glucose tolerance test. Elevated dietary NaCl significantly increased mean arterial pressure (MAP) in the uninephrectomized, but not the sham-operated groups. Acute administration of prazosin resulted in a significantly greater reduction in MAP of both hypertensive groups than of their normotensive counterparts. Moreover, clonidine caused a significant reduction in MAP of the hypertensive control rat but not in the normotensive controls. By contrast, both the hypertensive NIDD and the normotensive NIDD rats showed a similar reduction in MAP in response to clonidine administration. The data suggest that the combination of uninephrectomy and dietary NaCl excess confers hypertension on the NIDD rat. Moreover, enhancement of the sympathetic pathway plays an important role in the regulation of arterial pressure in the hypertensive NIDD rat. PMID- 10826409 TI - Lead promotes hydroxyl radical generation and lipid peroxidation in cultured aortic endothelial cells. AB - Early studies by our group have shown that lead-induced hypertension (HTN) is closely related to enhanced activity of reactive oxygen species (ROS). In addition, we have found indirect evidence that hydroxyl radical may be the most likely culprit in lead-exposed animals. In the present study, rat aortic endothelial cells were incubated in the presence of 0, 0.01, 0.1, 0.5, and 1.0 ppm lead acetate for 1, 24, and 48 h. At the conclusion of the incubation period cells were harvested and the media were collected. Lipid peroxidation products were measured as malondialdehyde-thiobarbituric acid (MDA-TBA) in the medium and hydroxyl radical was measured as 2,3-dihydroxybenzoic acid (2,3 DHBA) in the cells. After exposure to lead for 48 h, MDA-TBA generation and 2,3 DHBA formation were significantly increased. These data clearly demonstrate that lead exposure promotes hydroxyl radical generation and induces oxidative stress in isolated endothelial cells, mimicking the effects observed in lead-exposed animals. Enhanced inactivation of endothelium-derived nitric oxide by locally produced oxygen free radicals could contribute to endothelial dysfunction and HTN in lead exposed animals. PMID- 10826408 TI - Blood pressure and metabolic changes during dietary L-arginine supplementation in humans. AB - Dietary L-arginine supplementation has been proposed to reverse endothelial dysfunction in such diverse pathophysiologic conditions as hypercholesterolemia, coronary heart disease, and some forms of animal hypertension. In particular, chronic oral administration of L-arginine prevented the blood pressure rise induced by sodium chloride loading in salt-sensitive rats. To investigate the effects of L-arginine-rich diets on blood pressure and metabolic and coagulation parameters we performed a single-blind, controlled, crossover dietary intervention in six healthy volunteers. The subjects (aged 39+/-4 years, body mass index [BMI] 26+/-1 kg/m2, mean +/- SEM) received, in random sequence, three different isocaloric diets, each for a period of 1 week (Diet 1: control; Diet 2: L-arginine enriched by natural foods; Diet 3: identical to Diet 1 plus oral L arginine supplement). Sodium intake was set at a constant level (about 180 mmol/day) throughout the three study periods. A blood pressure decrease was observed with both L-arginine-rich diets (Diet 2 v 1, SBP: -6.2 mm Hg [95% CI: 0.5 to -11.8], DBP: -5.0 mm Hg [-2.8 to -7.2]; Diet 3 v 1, SBP: -6.2 mm Hg [-1.8 to -10.5], DBP: -6.8 mm Hg [-3.0 to -10.6]). A slight increase in creatinine clearance (P = .07) and a fall in fasting blood glucose (P = .008) occurred after Diet 3 and, to a lesser extent, after Diet 2. Serum total cholesterol (P = .06) and triglyceride (P = .009) decreased and HDL cholesterol increased (P = .04) after Diet 2, but not after Diet 3. These results indicate that a moderate increase in L-arginine significantly lowered blood pressure and affected renal function and carbohydrate metabolism in healthy volunteers. PMID- 10826410 TI - Role of the sympathetic nervous system during the development of obesity-induced hypertension in rabbits. AB - We have previously reported that weight gain induced by high-fat diet (HFD) leads to an increase in mean arterial pressure (MAP, +14%) and heart rate (HR, +31%) in the adult rabbit. In the present study, we tested the hypothesis that an increased activity of the sympathetic nervous system may contribute to the development of obesity-induced hypertension. A combination of alpha- and beta adrenergic blockers (terazosin + propranolol) was chronically administered to rabbits housed in metabolic cages for continuous monitoring of arterial pressure by telemetry, 24 h a day. After 2 weeks of adrenergic blockade under control diet, animals were switched to HFD for the next 6 weeks. HFD induced a progressive increase in body weight, but no increase in mean arterial pressure (+0.2+/-2.5%) and a slight increase in heart rate (+14+/-3%). Time-control animals fed normal diet showed no changes in MAP or HR with long-term alpha- and beta-adrenergic blockade. Our results indicate that the activation of the sympathetic nervous system may play an important role in the pathogenesis of obesity-induced hypertension. PMID- 10826411 TI - White coat hypertension in type 1 diabetic patients without nephropathy. AB - The aim of our study was to determine the prevalence of white coat hypertension (WCH) in type 1 diabetic patients. Therefore, ambulatory blood pressure monitoring (ABPM) and 24-h urinary albumin excretion (UAE) were determined in 47 patients with type 1 diabetes mellitus (27 with new diagnosis of hypertension by office blood pressure (BP) measurement and 20 with normotension). WCH was diagnosed in 20 patients (74%). Patients with WCH presented higher values of systolic and diastolic BP and UAE than normotensive patients. The results indicate that in type 1 diabetes mellitus WCH is very frequent. Thus, WCH may represent a potential risk for the development of diabetic complications, mainly diabetic nephropathy. PMID- 10826412 TI - Effects of vitamin E on clinic and ambulatory blood pressure in treated hypertensive patients. Collaborative Group of the Primary Prevention Project (PPP)--Hypertension study. AB - A randomized controlled open trial studied the effect of vitamin E supplementation (300 mg/day) on clinic and 24-h ambulatory blood pressure (BP) in 142 treated hypertensive patients. After 12 weeks, clinic BP decreased whether or not patients were randomized to vitamin E. Ambulatory BP showed no change in systolic BP and a small decrease in diastolic BP (-1.6 mm Hg, 95% confidence intervals from -2.8 to -0.4 mm Hg), approaching statistical significance in comparison to the control group (P = .06). Vitamin E supplementation thus seems to have no clinically relevant effect on BP in hypertensive patients already under controlled treatment. PMID- 10826413 TI - Hypertension articles in primary care journals. PMID- 10826414 TI - Natural antibodies and blood pressure hormones. PMID- 10826415 TI - Management of postoperative peritonitis after anterior resection: experience from a referral intensive care unit. AB - PURPOSE: Anastomotic leakage is the main cause of death after anterior resection. If it causes a single abscess, it may be successfully cured by percutaneous drainage, but in case of extensive peritoneal infection (multiple abscesses and generalized peritonitis), it is associated with a 40 to 60 percent mortality. This study aimed at evaluating aggressive, one-stage surgical management in such cases. METHODS: All patients referred to our surgical intensive care unit during the past ten years with generalized, multilocular, intra-abdominal sepsis after anterior resection were reviewed. There were 32 patients, with a mean age of 65 years, among which 15 (47 percent) were referred from other institutions. The mean Acute Physiology and Chronic Health Evaluation II score on admission was 18. All patients underwent a laparotomy with complete peritoneal exploration, intraoperative lavage, fecal diversion, capillary drainage of the pelvis excluding the rectal stump or the leaking anastomosis from the peritoneal cavity, and primary closure of the abdomen. A Hartmann's operation was done in 22 cases, and conservation of the anastomosis with proximal colostomy was done in 10 cases. The choice was based on the size of the leak, the viability of the colon, and the site of the anastomosis. RESULTS: Four patients died (12 percent), and five patients (16 percent) had recurrent sepsis. When the anastomosis had been conserved, restoration of continuity was achieved in all cases. After Hartmann's operation 8 patients of 19 survivors kept a permanent stoma; 7 had undergone a low anterior resection. CONCLUSIONS: Extensive intra-abdominal infection after anterior resection may be efficiently controlled by a surgical approach combining peritoneal debridement, fecal diversion, and capillary drainage of the pelvis. Intestinal continuity may be restored after diversion stoma or Hartmann's procedure after high anterior resection. This is not the case after a Hartmann's operation after a low colorectal anastomosis, and this procedure should be avoided whenever possible. PMID- 10826416 TI - Are sphincter defects the cause of anal incontinence after vaginal delivery? Results of a prospective study. AB - PURPOSE: Anal incontinence affects approximately 10 percent of adult females. Damage to the anal sphincters has been considered as the cause of anal incontinence after childbirth in the sole prospective study so far available. The aims of the present study were to determine prospectively the incidence of anal incontinence and anal sphincter damage after childbirth and their relationship with obstetric parameters. METHODS: We studied 259 consecutive females six weeks before and eight weeks after delivery. They were asked to fill in a questionnaire assessing fecal incontinence. Anal endosonography (7-10 MHz) was then performed. Two independent observers analyzed internal and external anal sphincters. RESULTS: A total of 233 patients (90 percent) were assessed, of whom 31 had cesarean section. De novo sphincter defects were observed in 16.7 percent (14 percent external, 1.7 percent internal, and 1 percent both) in the postpartum period only after vaginal delivery. These disruptions occurred with the same incidence after the first and the second childbirth. Independent risk factors (odds ratio; 95 percent confidence interval) for sphincter defect were forceps (12; 4-20), perineal tears (16; 9-25), episiotomy (6.6; 5-17), and parity (8.8; 4 19) as revealed by multivariate analyses. The overall rate of anal incontinence was 9 percent and independent risk factors (odds ratio; 95 percent confidence interval) involved forceps (4.5; 1.5-13), perineal tears (3.9; 1.4-10.9), sphincter defect (5.5; 5-15), and prolonged labor (3.4; 1-11). Among these patients only 45 percent had sphincter defects. CONCLUSION: Anal incontinence after delivery is multifactorial, and anal sphincter defects account for only 45 percent of them. Primiparous and secundiparous patients have the same risk factors for sphincter disruption and anal incontinence. Because external anal sphincter disruptions are more frequent than internal anal sphincter damage, surgical repair should be discussed in symptomatic patients. PMID- 10826417 TI - Neostigmine: safe and effective treatment for acute colonic pseudo-obstruction. AB - BACKGROUND: Ogilvie's syndrome, or acute colonic pseudo-obstruction, is a common and relatively dangerous condition. If left untreated, it may cause ischemic necrosis and colonic perforation, with a mortality rate as high as 50 percent. Neostigmine enhances excitatory parasympathetic activity by competing with acetylcholine for attachment to acetylcholinesterase at sites of cholinergic transmission and enhancing cholinergic action. We hypothesized that neostigmine would restore peristalsis in patients with acute colonic pseudo-obstruction. METHODS: Twenty-eight patients at Fletcher Allen Health Care and The Cleveland Clinic Foundation were treated for acute colonic pseudo-obstruction with neostigmine 2.5 mg IV over 3 minutes while being monitored with telemetry. Mechanical obstruction had been excluded. RESULTS: Complete clinical resolution of large bowel distention occurred in 26 of the 28 patients. Time to pass flatus varied from 30 seconds to 10 minutes after administration of neostigmine. No adverse effects or complications were noted. Of the two patients who did not resolve, one had a sigmoid cancer that required resection and one patient died from multiorgan failure. CONCLUSION: This study supports the theory that acute colonic pseudo-obstruction is the result of excessive parasympathetic suppression rather than sympathetic overactivity. We have shown that neostigmine is a safe and effective treatment for acute colonic pseudo-obstruction. PMID- 10826418 TI - Laparoscopic restorative proctocolectomy: case-matched comparative study with open restorative proctocolectomy. AB - PURPOSE: A laparoscopic approach to restorative proctocolectomy is new and has not been compared recently with the traditional open procedure. By using prospectively gathered data, laparoscopic and open restorative proctocolectomy procedures in mucosal ulcerative colitis and familial adenomatous polyposis patients were compared by using a case-matched design. METHODS: Forty patients, composing 20 consecutive laparoscopic cases (13 mucosal ulcerative colitis, 7 familial adenomatous polyposis), were matched for age, gender, and body mass index with 20 open cases (13 mucosal ulcerative colitis, 7 familial adenomatous polyposis) performed during the same time period. Mucosal ulcerative colitis patients were also matched for severity of disease by using hemoglobin and albumin levels, whole blood count, and steroid dependency. A loop ileostomy was made in 12 of 13 laparoscopic mucosal ulcerative colitis patients, all open mucosal ulcerative colitis patients, and no familial adenomatous polyposis patients. RESULTS: The median age was 25 (range, 9-61) years. There were no intraoperative complications in either group and no conversions in the laparoscopic group. The operative times (median, range) were significantly longer in laparoscopic cases (330, 180-480 minutes) vs. open cases (230, 180-300 minutes), P < 0.001. Bowel function returned more quickly in laparoscopic cases (2, 1-8 days) vs. open cases (4, 1-13 days), P = 0.03; and the length of stay was shorter in laparoscopic cases (7, 4-14 days) vs. open cases (8, 6-17 days), P = 0.02. For diverted patients, the median length of stay was reduced by two days in laparoscopic cases (6, 4-14 days) vs. open cases (8, 6-17 days), P = 0.01. Complications occurred in 4 of 20 laparoscopic patients (3 obstruction/ileus and 1 pelvic abscess) and 5 of 20 open patients (2 obstruction and ileus, 1 each anastomotic leak and abscess, peptic ulceration, and episode of dehydration). CONCLUSIONS: Return of intestinal function and length of stay are reduced in the laparoscopic group compared with open group. A laparoscopic approach to restorative proctocolectomy has the potential of becoming an appealing alternative to conventional restorative proctocolectomy surgery. PMID- 10826419 TI - Severe steroid-unresponsive ulcerative colitis: outcomes of restorative proctocolectomy in patients undergoing cyclosporin treatment. AB - PURPOSE: The recent introduction of the immune suppressor cyclosporin for treatment of steroid-refractory ulcerative colitis has required surgeons to perform a colectomy in those patients who eventually fail this rescue treatment, thus raising questions as to the safety of surgery as performed in patients with a heavily manipulated immune system. To assess the rates of mortality and morbidity in this setting, we studied a cohort of consecutive patients who had surgery after failing cyclosporin for refractory ulcerative colitis at our center. METHODS: Between January 1991 and December 1996, 25 patients with ulcerative colitis underwent restorative proctocolectomy performed in three steps (21 patients) and in two steps (4 patients). Seventeen of the 25 patients (68 percent) were initial nonresponders to a dose of 2 mg/kg/day of intravenous cyclosporin and underwent surgery immediately, the remaining 8 (32 percent) relapsed as outpatients on oral cyclosporin and were readmitted for surgery. RESULTS: There was no operative mortality. Nine patients of the 25 developed postoperative (early) complications (36 percent). The three-step operation subset had a 28 percent complication rate, the two-step 75 percent. Three patients needed reoperation. A total of 11 patients (44 percent) reported with late complications: two patients required surgical treatment, one for obstruction and one for pouch-perianal fistula. Three cases of pouchitis were recorded. No patient required pouch removal. CONCLUSION: Given the absence of postoperative mortality and a low overall complication rate, restorative proctocolectomy can safely be performed in patients who fail rescue treatment with a dose of 2 mg/kg of cyclosporin for steroid-refractory ulcerative colitis. Corollary evidence in this article hints but does not prove that the three-step procedure is safer than the two-step operation. PMID- 10826420 TI - Randomized, controlled trial to compare the J-pouch and W-pouch configurations for ulcerative colitis in the maturation period. AB - PURPOSE: Proctocolectomy with ileal pouch-anal anastomosis has become the procedure of choice for the treatment of ulcerative colitis. Functional results may differ with different pouch designs. This randomized study aimed to evaluate the relative effectiveness of two-limb J and four-limb W reservoir designs in the so-called maturation period after ileostomy closure. METHODS: Twenty-four patients underwent ileal pouch-anal anastomosis for ulcerative colitis. Eleven were randomly assigned to the J-pouch group and 13 to the W-pouch group. Frequency of defecation and other functional data were collected at 4, 8, and 12 months after ileostomy closure. Maximum tolerated volume was assessed in the same period by a latex balloon inflated with water. Maximum resting anal pressure, maximum voluntary contraction, and the rectoanal inhibitory reflex were assessed in the preoperative period and at 4, 8, and 12 months after ileostomy closure. RESULTS: Frequency of defecation decreased from 4 to 12 months after ileostomy closure in both groups (P = 0.04), but patients with a W-pouch had significantly lower values than patients with J-pouches (P < 0.01). Night-time defecation (P = 0.04) and use of antidiarrheals (P = 0.04) were significantly lower for patients with a W-pouch. Maximum tolerated volume was greater in the W-pouch group throughout the whole period (P = 0.01). Maximum resting anal pressure, maximum voluntary contraction, and rectoanal inhibitory reflex did not differ between the study arms. CONCLUSION: Patients with W-pouch have better functional results than those with J-pouches in the "maturation period" after ileostomy closure. PMID- 10826421 TI - Should enteric fistulas from Crohn's disease or diverticulitis be treated laparoscopically or by open surgery? A matched cohort study. AB - PURPOSE: The aim of this study was to study a group of consecutive patients with enteric fistulas treated by laparoscopic surgery and to compare outcomes with a matched group of patients treated by open surgery. METHODS: The outcomes of 13 patients with Crohn's disease or sigmoid diverticulitis with enteric fistulas treated laparoscopically (Group I) were compared with 13 patients matched for age, weight, gender, diagnosis, and characteristics of fistulas and treated by conventional surgery (Group II) during the same period. RESULTS: No patient died postoperatively in either group. Mean operative time was 183 minutes in Group I vs. 154 minutes in Group II (P = 0.280). No significant difference was found between Groups I and II in the number of patients with major postoperative complications (3 vs. 5; P = 0.462), or postoperative stay (7.6 +/- 3.6 vs. 9.2 +/ 3 days; P = 0.239). Conversion to open laparotomy occurred in one (7.7 percent) patient from Group I. No patient required readmission for secondary surgery in Group I, and two patients were readmitted and underwent reoperation for complications in Group II (P = 0.462). CONCLUSIONS: The laparoscopic treatment of selected cases of enteric fistulas is safe. Although most good outcome trends favor the laparoscopic group, the study is inconclusive, because no statistical difference was demonstrated with regard to operative time, number of postoperative complications, readmission rate, and length of postoperative stay, most likely because of the small number of cases in each arm of the study. Study of a greater number of cases outside the learning curve of the laparoscopic surgeons would clarify this issue. Other outcomes, including cost, pain control, cosmesis, and return to activities of daily living, need to be included in the evaluation. PMID- 10826422 TI - Abdominal wall recurrence after colorectal resection for cancer. AB - PURPOSE: Disease recurrence in the abdominal wall from a primary colorectal cancer is a poorly studied and little understood phenomenon that has received renewed attention after the recognition of port site metastases in patients after laparoscopic colorectal resections. The purpose of the present study was to define the clinical, pathologic, and management issues in patients with abdominal wall metastases from colorectal cancer. METHODS: Patients presenting to Memorial Sloan-Kettering Cancer Center with a diagnosis of colorectal cancer were entered into a prospective database beginning in 1986. Review of this database showed that 31 patients presenting with recurrent disease in the abdominal wall were managed surgically at the institution between 1986 and 1998. RESULTS: A total of 31 patients (19 males) with a median age of 67 (range, 45-86) years presented with recurrent disease between 7 and 183 (median, 24) months after primary surgery. Primary tumors were located in the right colon in 17 patients, left colon in 2 patients, sigmoid colon in 7 patients, and rectum in 3 patients. Nineteen percent of primary tumors were perforated, 45 percent were poorly differentiated, 92 percent were transmural (T3 or T4), and 51 percent had lymph node metastases at presentation. Twenty-two patients presented with a symptomatic abdominal wall mass, whereas recurrence in the abdominal wall was found incidentally in 9 patients undergoing laparotomy. Four patients had isolated abdominal wall disease, whereas the remaining 27 were found to have associated intra-abdominal disease. Six patients who were left with residual intra-abdominal cancer after abdominal wall resection had a median survival time of four months. Twenty-five patients underwent a histologically complete resection of recurrence restricted to the abdominal wall alone (n = 4; median survival time, 18 months), abdominal wall and in continuity resection of adherent viscera (n = 15; median survival time, 12.5 months), or resection of abdominal wall and intra-abdominal recurrence at a distant site (n = 6; median survival time, 22 months, although only 1 patient remained alive with disease). The actual two-year and five-year disease-free survival rates were 16 and 3 percent, respectively. CONCLUSION: Abdominal wall metastases are often indicators of recurrent intra-abdominal cancer; however, aggressive resection in patients with disease restricted to the abdominal wall and associated adherent viscera can result in local disease control with little morbidity and no mortality. PMID- 10826423 TI - Pudendal nerve function in idiopathic fecal incontinence. AB - PURPOSE: The study was undertaken to determine whether idiopathic fecal incontinence in middle-aged and elderly females is likely to be a result of pudendal nerve damage (neurogenic incontinence) or merely a consequence of aging. METHODS: One hundred seventy-eight females over the age of 50 years with fecal incontinence were studied. The incontinence was classified as idiopathic because none of the patients related the incontinence to trauma (including obstetric trauma) or other events or diseases. All had an anal physiology examination, including determination of nerve conduction velocity of both pudendal nerves (pudendal nerve terminal motor latency). RESULTS: With a cutoff value of 2.4 msec, 79 percent (95 percent confidence limit, 73-85) had normal pudendal nerve terminal motor latency on both sides, 13 bilaterally prolonged latency (7 percent; 4-11), and 25 unilaterally prolonged latency (14 percent; 9-19). With a cutoff value of 2.2 msec, 66 percent (59-73) had normal latency on both sides, 15 percent (9-20) bilaterally prolonged pudendal nerve terminal motor latency, and 20 percent (14-26) unilateral prolongation. No relationship between the groups with normal, bilateral, or unilateral prolongation of pudendal nerve terminal motor latency and anal resting and squeeze pressure was found. Anal resting pressure decreased with increasing age (P < 0.05). CONCLUSION: Our data support the view that idiopathic fecal incontinence in the majority of females is likely to be a result of the aging process and that only a limited number may suffer from anal incontinence of neurogenic origin. Furthermore, unilateral prolongation of pudendal nerve terminal motor latency probably is without clinical significance. PMID- 10826424 TI - Laparoscopic suture rectopexy without resection is effective treatment for full thickness rectal prolapse. AB - PURPOSE: The study was undertaken to evaluate the role of laparoscopic suture rectopexy without resection as a safe and effective treatment for full-thickness rectal prolapse. METHOD: Data were prospectively collected and analyzed on 25 patients who underwent laparoscopic rectopexy without resection for full thickness rectal prolapse between October 1994 and July 1998. Four patients had conversions from laparoscopic to open surgery. Two patients had recurrent prolapse previously managed by Delorme's procedure. Another two patients had solitary rectal ulcer syndrome associated with their full-thickness rectal prolapse. There were a total of three males. Mean age was 72 (range, 37-89) years. The preoperative and postoperative course of each patient was followed up, with attention paid to first bowel movement, hospital stay, duration of surgery, fecal incontinence, constipation, recurrent prolapse, morbidity, and mortality. Follow-up was made by clinic appointments and, if necessary, by telephone review. RESULTS: Median follow-up period was 26 (range, 1-41) months. Mean duration of surgery was 96 (range, 50-150) minutes. Postoperatively, the median time for first bowel movement was four (range, 2-10) days. Median hospital stay was seven (range, 3-23) days. Overall, 15 patients (60 percent) either improved or remained unchanged with respect to continence. There was an improvement in 10 of 20 patients (50 percent) among those with continence Grade 2 or more (P < 0.05). Seven patients (28 percent) remained incontinent. No patient became more incontinent after surgery. Constipation, which was present in 9 patients (36 percent) preoperatively, affected 11 patients (44 percent) after rectopexy (P > 0.05; not significant). Postoperative morbidity included a port site hernia and deep venous thrombosis in one patient, a repaired rectal perforation, a retroperitoneal hematoma with prolonged ileus (1 case), and a superficial wound infection (1 case). One patient with solitary rectal ulcer syndrome in the laparoscopic surgery group remained unhealed despite resolution of the rectal prolapse after rectopexy and required abdominoperineal resection. Two patients (laparoscopic surgery = 1 and open surgery = 1) had severe constipation after surgery and both required loop colostomies. There were no cases of operative mortality or recurrent prolapse. CONCLUSION: Laparoscopic suture rectopexy without resection is both safe and effective in this frequently frail population and offers a minimally invasive approach that may have potential advantages for selected groups of patients with full-thickness rectal prolapse. PMID- 10826425 TI - Schistosomal pelvic floor myopathy contributes to the pathogenesis of rectal prolapse in young males. AB - PURPOSE: Rectal prolapse is common in young males in Egypt. The role of schistosomiasis in the pathogenesis of rectal prolapse is not clearly defined. The purpose of this work is to study changes in the pelvic floor muscles in patients of rectal prolapse associated with schistosomiasis. METHODS: This study included 33 male patients with rectal prolapse of whom 27 patients with schistosomiasis and 6 patients free from schistosomiasis. Biopsies were taken from the pelvic floor muscle during surgery. The prepared sections were examined for histopathologic structural changes, for ultrastructural changes (by using electron microscopy) and after immunohistochemical staining by using anti-IgG and anti-IgM antibodies. RESULTS: The muscles from the patients without schistosomiasis had no histologic or EM changes and showed negative staining for IgG and IgM. Myopathic changes were found in the group of patients with schistosomiasis, including increased variation in the fiber diameter in 66.6 percent of patients, degenerative changes in 59.26 percent of patients, fiber splitting and fragmentation in 44.4 percent of patients, and endomysial fatty changes in 55.5 percent of patients. Ultrastructural study revealed starting loss of striation and margination of the nucleus in 70.37 percent of patients, distorted myofibrillar pattern in 51.85 percent of patients, disturbed endoplasmic reticulum and increased glycogen granules in all patients, the mitochondria are irregularly arranged with electron dense matrix in 40.74 percent of patients, and prominent nuclear sap in 13.72 percent of patients. The muscles of all the schistosomal patients showed a positive cytoplasmic staining for immunoglobulin G, of them only 12 patients showed positive staining for IgM. CONCLUSION: Patients with schistosomiasis suffer from pelvic floor myopathic changes that may contribute to the pathogenesis of rectal prolapse in young males. The immunohistochemical findings suggest immunologic mechanism for this myopathy. PMID- 10826426 TI - Quality of life with a temporary stoma: ileostomy vs. colostomy. AB - PURPOSE: The hypothesis is that the impact of a temporary stoma on a patient's daily life is determined by complications and related stoma care problems. METHOD: A prospective clinical trial was performed, studying complications and social well-being of 37 patients with loop ileostomy and 39 patients with a loop colostomy (randomly assigned comparison). Patients were categorized according to degree of social restriction. The association between the degree of social restriction and the presence of stoma care problems and complications was assessed. Follow-up was scheduled every three months until the stoma was closed (94 percent). RESULTS: There is no relation between stoma type (ileostomy or colostomy) and degree of social restriction (chi-squared test, P = 0.42). The more stoma care problems or complications seen, the higher the degree of social restriction: significantly more stoma care problems were seen in the completely isolated group of patients when compared with the patients who were less socially restricted (Spearman correlation coefficient 1 = 0.35, P = 0.003). Especially stoma leakage, peristomal skin irritation, dietary prescriptions, retraction, and prolapse of the stoma have significant impact on the patient's daily life. CONCLUSION: Stoma surgery has a great influence on a patient's daily life. There is a clear relation between the number of stoma care problems and the degree of social restriction. Follow-up of stoma patients under close surveillance of stoma care nurse to minimize stoma care problems and a careful surgical technique are advocated for good stoma care. PMID- 10826427 TI - Survival after percutaneous, image-guided, thermal ablation of hepatic metastases from colorectal cancer. AB - PURPOSE: One-year, two-year, three-year, and four-year survival rates and median survival time for patients with inoperable liver metastases from colorectal cancer is 32, 10, and 3 percent and 7.4 to 11 months, respectively. Systemic chemotherapy produces a modest improvement to 48, 21, and 3 percent and 12 months, respectively. Regional chemotherapy produces a further improvement to 64, 25, and 5 percent and 15 to 17 months, respectively. For those with operable disease, hepatic resection survival rates are 90, 62, 48, and 40 percent, respectively, and survival time is 33 months. Thermal ablation is effective in producing necrosis in liver metastases. We report the impact on survival in 69 patients treated from 1993 to 1997, with follow-up to 1998. METHODS: Sixty-nine patients, 50 male, mean age 60 (range, 33-87) years were treated. Liver resection was not feasible because of disease extent in the liver, extrahepatic disease or concurrent medical conditions. The average number of liver metastases was 2.9 (range, 1-16), the mean maximal diameter was 3.9 (range, 1-8) cm, and the mean initial total liver tumor volume was 47 (range, 1-371) ml. Eighteen (26 percent) had undergone previous hepatic resection. Sixty-two of 67 (93 percent) received chemotherapy at some stage. Twenty (29 percent) had extrahepatic disease. RESULTS: One-year, two-year, three-year, and four-year survival rates and median survival time from liver metastasis diagnosis was 90, 60, 34, and 22 percent and 27 months, respectively. Forty of 69 (58 percent) developed new liver metastases, and 23 of 69 (33 percent) developed new extrahepatic disease. Of a subgroup of 24 patients with less than four metastases, <5 cm diameter, treated after January 1995, the median survival time was 33 months from first thermal ablation vs. 15 months for the remainder (P = 0.0004). Major morbidity occurred in 3.2 percent, minor morbidity occurred in 12 percent, and there was one periprocedural death. CONCLUSIONS: Thermal ablation therapy improves survival in patients with inoperable but limited liver metastases. This is an improvement on the natural history of the disease and published chemotherapy results. Recent and ongoing technical refinements, not reflected in these results, are expected to further improve survival. PMID- 10826428 TI - Transanal endoscopic microsurgery: experience with 75 rectal neoplasms. AB - PURPOSE: The aim of this study was to describe a single institution's experience with transanal endoscopic microsurgery in patients with benign and malignant rectal tumors. PATIENTS: Between January 1992 and April 1998, 75 patients with a mean follow up of 38 months, underwent transanal endoscopic microsurgery excision of benign (46) or malignant (29) rectal tumors, located 3 to 18 cm from the dentate line. RESULTS: A total of 3 of 46 (6.5 percent) patients with benign tumors underwent conversion to radical surgery owing to tumor size. During the follow-up period, benign tumor recurrence was observed in four (9 percent) patients, three of whom were managed by repeat transanal endoscopic microsurgery, whereas one required radical surgery. Histologic staging of malignant tumors was T1 (10), T2 (10), and T3 (9). Seven patients with either inadequate resection margins or T3 tumors were complimented with radical surgery. Of the remaining 22 patients, 11 received adjuvant radiation therapy whereas 11 had no further treatment. Four (18 percent) had recurrent disease, which was managed by repeat transanal endoscopic microsurgery in two, radical surgery in one, and laser ablation in one. No cancer-related deaths were observed during the follow-up period. There was one operative mortality in a cardiac-crippled patient. Postoperative complications were mainly of a minor character and included fever, urinary retention, and bleeding; none of which required reintervention. Rectourethral fistula developed in one patient who underwent repeat transanal endoscopic microsurgery excision for a T3 malignancy. Fecal soiling was transient in three patients and persisted in two. CONCLUSION: Transanal endoscopic microsurgery excision is a safe and precise technique that is well tolerated even in high operative risk patients. Transanal endoscopic microsurgery may become a procedure of choice for benign rectal tumors and selected early malignant neoplasms. PMID- 10826429 TI - Bowel preparation with oral polyethylene glycol electrolyte solution vs. no preparation in elective open colorectal surgery: prospective, randomized study. AB - PURPOSE: Efficient mechanical bowel preparation has been regarded as essential in preventing postoperative complications of colorectal surgery, but the necessity of bowel cleansing has been disputed recently. The aim of this study was to evaluate the outcome of elective colorectal surgery in patients with or without bowel preparation. METHODS: Altogether, 267 consecutive adult patients admitted for elective open colorectal surgery were randomly assigned either to the bowel preparation group with oral polyethylene glycol electrolyte solution (138 patients) or no preparation group (129 patients). Patients who were unable to drink polyethylene glycol electrolyte solution, those who had had bowel preparation within the previous week, and patients not needing opening of the bowel were excluded. Routine colorectal surgery was undertaken, and infectious and other complications were registered daily. Late complications were checked up one to two months after surgery. RESULTS: No deaths were recorded, and 76 percent of the patients in the polyethylene glycol electrolyte solution group and 81 percent in the unprepared group recovered without complication. Anastomotic leaks occurred in 4 percent of the polyethylene glycol electrolyte solution patients and in 2 percent of the other cases, and other surgical site infections occurred in 6 and 5 percent, respectively. None of the differences was statistically significant. There was no difference in restoration of bowel function. The median postoperative stay was eight days in both groups. CONCLUSIONS: Preoperative bowel preparation seems to offer no benefit in elective open colorectal surgery. PMID- 10826430 TI - Intraoperative lower extremity compartment pressures in lithotomy-positioned patients. AB - PURPOSE: Measurement of anterior leg compartment pressures in eight patients (16 limbs) who were positioned in the lithotomy position for prolonged periods of time while undergoing colorectal intra-abdominal surgery. METHODS: Anterior leg compartment pressures were measured in eight patients (16 limbs) by using a slit catheter, whereas subjects were positioned in the lithotomy position for prolonged periods of time while undergoing colorectal intra-abdominal surgery. RESULTS: Compartment pressures had minor elevations after initial lithotomy positioning, and gradually increased over time. Levels elevated to 30 mmHg at an average of five (range, 3.5 to 6) hours. The maximum recorded leg compartment pressure was 70 mmHg. The addition of Trendelenburg positioning was noted to consistently increase compartment pressures. All pressures returned to less than 10 mmHg shortly after removing the limb from the stirrups and placing the limb supine. No patients developed clinical evidence of compartment syndrome. CONCLUSIONS: Leg anterior compartment pressures rise when limbs are placed in the lithotomy position for prolonged periods of time. The rise in pressure is increased with the addition of Trendelenburg positioning. Anterior compartment pressures reached a threshold of 30 mmHg at an average of five hours. The results of this study suggest that lithotomy positioning of the lower extremities has the potential to initiate leg compartment syndrome when the period of positioning approaches five hours. Removing the limbs from the stirrups and placing them in the supine position allows the pressure in the compartments to return to normal. PMID- 10826431 TI - Dermal island-flap anoplasty for transsphincteric fistula-in-ano: assessment of treatment failures. AB - PURPOSE: The aim of this study was to assess the treatment failures of island flap anoplasty for fistula-in-ano, a procedure designed to treat fistula without sphincter division. METHODS: Data concerning all patients having dermal island flap anoplasty for the treatment of transsphincteric fistula were reviewed. Variables assessed were age, gender, radial fistula location, cause, Crohn's disease, previous fistula operations, other complicating illnesses, internal sphincter closure, simultaneous use of fibrin adhesive injection, and use of combined dermal and rectal flap for large fistulas. Postoperative data collected included persistence of the distal tract, recurrence of the fistula, and treatment of the recurrence. Recurrence (or persistence) of the fistula was the dependant variable and each risk factor for recurrence was assessed using chi squared analyses. RESULTS: Seventy-three flaps were performed in 65 individuals. Recurrence developed 17 times in 13 individuals. Recurrence was more likely to occur in males, patients who have had previous treatment of fistulas, patients with large fistulas requiring combined flaps, and patients who had simultaneous fibrin glue injection. Patients with Crohn's disease and individuals having internal sphincter closure had fewer recurrences. Factors reaching statistical significance included closure of the internal sphincter, the use of fibrin glue, and cause of the fistula. CONCLUSION: No specific anatomic or demographic characteristic is sufficiently associated with failure to exclude any patient from the operation. Closure of the internal sphincter should be done as part of the procedure and fibrin glue injection should not be done simultaneously. PMID- 10826432 TI - Clinical, endoscopic, and histologic spectrum of nonsteroidal anti-inflammatory drug-induced lesions in the colon. AB - PURPOSE: It has become increasingly clear that nonsteroidal anti-inflammatory drugs may cause damage not only to the upper gastrointestinal tract but also to the small and large intestine. Although the colon may be readily investigated by endoscopy, drug-induced lesions are not well known, probably because they are considered to occur only rarely. In the present study we describe endoscopic, histologic, and gross characteristics of nonsteroidal anti-inflammatory drug induced colonic damage. Furthermore, pathogenetic mechanisms and therapeutic options are discussed. METHODS: The histories of all patients diagnosed as having nonsteroidal anti-inflammatory drug colitis during the last two years at the department of gastroenterology or the department of pathology at our hospital were reviewed. Endoscopic, histologic, and gross pathologic findings were systematically recorded. In addition, data on duration and type of nonsteroidal anti-inflammatory drug intake and time from onset of symptoms to diagnosis were collected. Therapy and outcome of our patients, if available, are reported. RESULTS: During the study period 11 patients were diagnosed as having nonsteroidal anti-inflammatory drug colitis. Most patients presented with diarrhea with or without blood loss and complained about diffuse abdominal pain. Endoscopy revealed flat ulcers in the entire colon being more severe in the right colon in the three cases with acute onset of diarrhea. In four cases concentric "diaphragm-like" strictures were seen, all located in the right colon. In the remainder endoscopy showed nonspecific erosions and was normal in one patient. Histology revealed findings similar to ischemic colitis. Additionally, in two cases collagenous colitis was found. Diclofenac slow release was the most commonly involved drug. The median time from onset of symptoms to diagnosis was 1.8 (range, 0-11.5) years. CONCLUSIONS: Nonsteroidal anti-inflammatory drug colitis is a clinically significant disease, which may present with diarrhea, anemia, and nonspecific abdominal complaints. Careful history taking, together with awareness of endoscopic and histologic findings, allows a timely diagnosis of this disease. PMID- 10826433 TI - Long-term clinical course and prognostic factors in intestinal Behcet's disease. AB - PURPOSE: The present study was aimed at evaluating the long-term course of intestinal Behcet's disease and determining predictive factors of prognosis. METHODS: This report is a retrospective study based on the records of 43 patients with intestinal Behcet's disease. The mean follow-up duration was 73 +/- 60 months. We evaluated the efficacy of medical treatment for the intestinal lesion at initial eight weeks. The cumulative probabilities were calculated by using Kaplan-Meier method, and the results were compared by using the log-rank test. RESULTS: Sixteen patients (38 percent) achieved a complete remission of intestinal lesions eight weeks after medical treatment had begun. The patients who achieved a complete remission had a lower probability of receiving an operation than those who had not (13 percent at 2 and 5 years vs. 36 and 43 percent, respectively; P = 0.028). The recurrence probability of intestinal lesions was 25 percent at two years and 49 percent at five years after complete remission with medical treatment. Patients who had a history of intestinal perforation or fistula had a higher probability of recurrence after operation than those without such history (59 vs. 33 percent at 2 years; 88 vs. 57 percent at 5 years; P = 0.020). Patients who had taken azathioprine had a lower probability of receiving reoperation than those who did not (7 vs. 25 percent at 2 years; 25 vs. 47 percent at 5 years; P = 0.035). The length of ileal resection and whether hemicolectomy was performed had no significant effect on the recurrence or reoperation rate. CONCLUSIONS: Intestinal Behcet's disease frequently requires a surgical treatment and has a high recurrence rate. The patients who achieved a complete remission with medical treatment, who had no history of intestinal perforation, and who received azathioprine after operation showed better clinical courses. Resection of a short segment of bowel would be a more appropriate surgical procedure. PMID- 10826434 TI - Simple and effective surgical treatment of pilonidal sinus: asymmetric excision and primary closure using suction drain and subcuticular skin closure. AB - PURPOSE: This study was planned to evaluate prospectively the results of 112 pilonidal sinus cases treated surgically by using asymmetric excision and primary closure with suction drain and subcuticular skin closure. It is aimed at elimination of the causative factors of pilonidal sinus. METHOD: The patient's age, profession, weight and height, symptoms and signs, duration of symptoms, previous treatments, operation time and cost, hospital stay, return to normal activity, complications, pathologic and microbiologic examinations, and recurrences were noted. All pilonidal sinus cases except pilonidal abscess and extensive gluteal involvement were treated surgically. The procedure consists of an eccentric, elliptical excision of the affected tissue, mobilization of the flap to the sacrococcygeal fascia and the suturing of its edge to the lateral one. Penrose drains were placed in the first eight (7.14 percent) cases, but suction drains were placed in others. The cases were followed up for a mean of 2.4 years. RESULTS: Twenty-eight (25 percent) cases had undergone previous operative procedures. Of 112 patients 106 (94.6 percent) were male. Mean age was 22.1 years. Mean history of disease was 4.2 years. The overall complication rate was 7.14 percent. Two (1.8 percent) wound infections, two wound breakdowns, three (2.7 percent) collections, and one (0.9 percent) recurrence were recorded. The collections were reduced to zero after first eight cases by using a suction drain. Sixty-eight of the patients (60.7 percent) had body weight over 90 kg, and the mean body mass index was 24.8. The mean hospital stay was 2.6 days, and the mean time off work was 12.4 days. The average healing time was 13.2 days. There were no anesthetic or surgical deaths. CONCLUSION: The natal cleft is flattened and the incision scar and the incision line is transferred from the midline to the lateral side by performing the asymmetric excision and primary closure, and thus the essential cause of pilonidal sinus is eliminated. The procedure is simple, the complications and recurrences are very low, and it is seen to be an excellent procedure in the surgical treatment of uncomplicated pilonidal sinus disease. PMID- 10826435 TI - Cecal necrosis: infrequent variant of ischemic colitis. Report of five cases. AB - PURPOSE: Spontaneous nonocclusive ischemic colitis involving only the right colon is an infrequent occurrence. Because this problem is less recognized than its counterpart involving the left colon, the correct diagnosis may not be considered. The purpose of this article was to describe the presentation and management of this unusual clinical problem. METHODS: Five cases of nonocclusive ischemic cecal necrosis are described. Four of the patients presented with right sided abdominal pain, tenderness, and leukocytosis. The preoperative diagnosis was incorrect in all patients, although cecal necrosis was considered in one. Two patients were thought to have. appendicitis, two were thought to have carcinoma, and one was thought to have a perforated viscus. Each patient underwent a right hemicolectomy and four survived. RESULTS: Each of the patients had ischemic cecal necrosis without evidence of emboli or vasculitis. Although cecal gangrene may occur after systemic hypotension, no such event preceded these patients' presentation. We believe that the patients we treated had a form of nonocclusive ischemic colitis, which occasionally affects only the right colon. CONCLUSION: Ischemic necrosis of the cecum is an infrequent variant of ischemic colitis that should be considered in the differential diagnosis of the elderly patient presenting with right lower quadrant pain. PMID- 10826436 TI - Successful resection of an ileoanal pouch mesenteric desmoid without pouch sacrifice: report of a case. AB - PURPOSE: Intra-abdominal desmoid tumors are associated with familial adenomatous polyposis and may compromise ileoanal pouch function after restorative proctocolectomy, particularly if the pouch mesentery is involved. This usually necessitates pouch excision. The aim of this report was to describe the first known case of pouch salvage after surgery for a desmoid tumor arising from the pouch mesentery. METHODS: The management of a desmoid tumor involving an ileoanal pouch in a 21-year-old female is described and the literature reviewed. RESULTS: The tumor was successfully excised completely after failed medical therapy, and pouch function was fully preserved. CONCLUSIONS: This is a unique case highlighting the possibility of pouch salvage when affected by desmoid tumor. PMID- 10826437 TI - Transperitoneal drainage for a large cystic degeneration after regression of an intra-abdominal desmoid tumor. AB - A new technique is described for treating a large cystic degeneration after regression of an intra-abdominal desmoid tumors in patients with familial adenomatous polyposis. A cysto-peritoneal shunt is made for the relief of large cystic degeneration by creating a channel between the mesenteric cyst and the peritoneal cavity by means of a silicone tube to facilitate transperitoneal drainage. PMID- 10826438 TI - The farthest 3' distal end APC mutation identified in attenuated adenomatous polyposis coli with extracolonic manifestations. PMID- 10826439 TI - Nonoperative management of chronic anal fissure. PMID- 10826440 TI - Familial adenomatous polyposis: extending the role of dye-spray at colonoscopy. PMID- 10826441 TI - Femoral nerve neuropraxia after abdominoperineal resection. PMID- 10826442 TI - "Somatoanal" reflex or "thermosphincteric" reflex? PMID- 10826443 TI - Homeopathy in cancer care. PMID- 10826444 TI - Statistical analysis of the effect of high dilutions of arsenic in a large dataset from a wheat germination model. AB - This paper describes the statistical analysis of a series of experiments using a simple biological model (wheat germination in vitro), where a large number of wheat seeds were treated with homeopathic potencies of Arsenic trioxide. Some potencies, such as As2O3 40x, 42x and 45x, have repeatedly shown a significant stimulating effect on germination compared to controls, whereas As2O3 35x has a significant inhibiting effect. In some experiments the seeds were stressed before the experiment with a sublethal dose of the same substance. We performed a statistical analysis, both for stressed and non-stressed seed groups, using Poisson distribution as a suitable model for representing the number of non germinated seeds in a standard experiment with 33 seeds in the same Petri dish. Finally, we have considered the most repeated potencies (30x and 45x), computing the sample odds ratio (OR) and a 95% confidence interval (CI) for the population OR. Our results show significant reproducible effects of some As2O3 decimal potencies, particularly As2O3 45x. In stressed seeds, even decimal potencies of water seem to give significant results compared to control, whereas high dilutions of As2O3 without potentization never show significant effects. PMID- 10826445 TI - Patient benefit survey: Tunbridge Wells Homoeopathic Hospital. AB - We report an outcome study concerning patients treated by homeopathic medicine at the Tunbridge Wells Homoeopathic Hospital for the whole of 1997. The study aimed to assess: (a) the range of diagnoses presented by patients, and (b) patients' own impressions of benefit. 1372 questionnaires were completed by patients after consultations to record their impressions of the effects of homeopathic treatment. Patients were asked to score their responses on a + 3 to - 3 scale. The three main diagnostic groups were, dermatology, musculo-skeletal disorders, and malignant disease, especially carcinoma of the breast. Overall, 74% of patients recorded positive benefits, with 55% recording scores of + 3 or + 2. PMID- 10826446 TI - Altered solution structure of alcoholic medium of potentized Nux vomica underlies its antialcoholic effect. AB - Nux vomica 30c, 200c and 1000c were administered orally to three batches of albino mice for three days. Six hours after the last dose on the third day the mice were injected i.p. with ethanol 4g/kg body wt. They lost their righting reflex and lay motionless apparently sleeping due to alcohol. Mice treated with three potencies of Nux vomica regained their righting reflex more quickly than the corresponding untreated controls. Each of the three batches of mice was tested twice for ethanol sedation, once with a potency of Nux vomica and another time with a placebo control. The time interval between drug treatment and control was 10 days. NMR spectra of Nux 30, Nux 200, Nux 1000, alcohol 30, alcohol 30 (unagitated) and 90% alcohol showed significant difference from each other with respect to the spin-lattice relaxation time (T1) of the deuterium nuclei. This gives a measurable physical basis of the effective high potencies of Nux vomica. PMID- 10826447 TI - A new homeopathic approach to neoplastic diseases: from cell destruction to carcinogen-induced apoptosis. AB - Neoplastic diseases are now among the most commonly seen conditions. Orthodox, non-surgical approaches, including chemotherapy and radiotherapy, have variable results, but many adverse affects that limit their use. These are sometimes the direct cause of death. More patients are choosing alternative treatments, mainly the homeopathic and herbal-nutrition approach. Homeopathy does not have highly effective remedies for cancer in its literature, and has been limited to palliating the adverse effects of chemo/radiotherapy. Research into substances that can produce neoplastic diseases (carcinogens), may lead to them being used to treat the cancer they cause, according to the principle of similarity. I have used ultra-low doses (1 x 10(-10) to 10(-12) molar) of chemical carcinogens for 3 24 months, which have been given to cancer patients, usually in conjunction with conventional treatment. Using this procedure, complete remission or life extension has been achieved for some cancer cases. Three clinical cases are presented: a man with undifferentiated lung cancer; a child with an astrocytoma and a woman with leiomyosarcoma. PMID- 10826448 TI - The toxicology of Trachinus vipera: the lesser weeverfish. AB - The biology and toxicology of the lesser weeverfish, Trachinus vipera, is presented in a homeopathic repertory fashion to supplement the existing materia medica. PMID- 10826449 TI - 20 years ago: the British Homoeopathic Journal, April 1980. PMID- 10826450 TI - Homeopathy in Cuba. PMID- 10826451 TI - Management of community-acquired pneumonia in the era of pneumococcal resistance: a report from the Drug-Resistant Streptococcus pneumoniae Therapeutic Working Group. AB - OBJECTIVE: To provide recommendations for the management of community-acquired pneumonia and the surveillance of drug-resistant Streptococcus pneumoniae (DRSP). METHODS: We addressed the following questions: (1) Should pneumococcal resistance to beta-lactam antimicrobial agents influence pneumonia treatment? (2) What are suitable empirical antimicrobial regimens for outpatient treatment of community acquired pneumonia in the DRSP era? (3) What are suitable empirical antimicrobial regimens for treatment of hospitalized patients with community-acquired pneumonia in the DRSP era? and (4) How should clinical laboratories report antibiotic susceptibility patterns for S pneumoniae, and what drugs should be included in surveillance if community-acquired pneumonia is the syndrome of interest? Experts in the management of pneumonia and the DRSP Therapeutic Working Group, which includes clinicians, academicians, and public health practitioners, met at the Centers for Disease Control and Prevention in March 1998 to discuss the management of pneumonia in the era of DRSP. Published and unpublished data were summarized from the scientific literature and experience of participants. After group presentations and review of background materials, subgroup chairs prepared draft responses, which were discussed as a group. CONCLUSIONS: When implicated in cases of pneumonia, S pneumoniae should be considered susceptible if penicillin minimum inhibitory concentration (MIC) is no greater than 1 microg/mL, of intermediate susceptibility if MIC is 2 microg/ mL, and resistant if MIC is no less than 4 microg/mL. For outpatient treatment of community-acquired pneumonia, suitable empirical oral antimicrobial agents include a macrolide (eg, erythromycin, clarithromycin, azithromycin), doxycycline (or tetracycline) for children aged 8 years or older, or an oral beta-lactam with good activity against pneumococci (eg, cefuroxime axetil, amoxicillin, or a combination of amoxicillin and clavulanate potassium). Suitable empirical antimicrobial regimens for inpatient pneumonia include an intravenous beta-lactam, such as cefuroxime, ceftriaxone sodium, cefotaxime sodium, or a combination of ampicillin sodium and sulbactam sodium plus a macrolide. New fluoroquinolones with improved activity against S pneumoniae can also be used to treat adults with community-acquired pneumonia. To limit the emergence of fluoroquinolone-resistant strains, the new fluoroquinolones should be limited to adults (1) for whom one of the above regimens has already failed, (2) who are allergic to alternative agents, or (3) who have a documented infection with highly drug-resistant pneumococci (eg, penicillin MIC > or =4 microg/mL). Vancomycin hydrochloride is not routinely indicated for the treatment of community-acquired pneumonia or pneumonia caused by DRSP. PMID- 10826452 TI - Does aspirin attenuate the beneficial effects of angiotensin-converting enzyme inhibition in heart failure? AB - Ischemic heart disease is the most common underlying cause of congestive heart failure, and thus aspirin (acetylsalicylic acid [ASA]) and angiotensin-converting enzyme (ACE) inhibitors are commonly used together for treatment in this setting. The issue of possible attenuation of the effect of ACE inhibitors by ASA has been an area of intense debate. Currently, it is perceived that a significant part of the beneficial effect of ACE inhibitors is related to augmentation of bradykinin levels, which among other effects stimulate the release of prostacyclin. Aspirin, on the other hand, inhibits the production of prostacyclin by blocking cyclooxygenase. Prostaglandins play an important endogenous vasodilatory role and counteract the enhanced peripheral vasoconstriction state in congestive heart failure. Thus, the counteracting effect of ASA on the augmentation of prostacyclin synthesis by ACE inhibitors could result in a potential reduction of the beneficial effects of the ACE inhibitor's and could be of great importance. This article reviews reports from large clinical trials pertaining to this issue and relates their findings to the currently available theoretical bases for support of the counteracting effect of ASA on augmentation of prostacyclin synthesis by ACE inhibitors. The clinical implications of such an interaction are discussed. PMID- 10826453 TI - Charges for medical care at different hospitals. AB - BACKGROUND: The United States has a high proportion of people without health insurance (15%) and a low proportion of people without employment (5%), resulting in millions who lack insurance but have some ability to pay. We tested whether hospitals charge similar prices for well-specified elective services to individuals paying out-of-pocket for medical care. METHODS: We surveyed the 2 largest general hospitals from every large city (population >500 000) in the United States and Canada. At each hospital we evaluated 5 diagnostic, 7 therapeutic, and 3 nonclinical services to determine the total charge to patients who pay directly. RESULTS: Overall, 66 hospitals were included (average, 758 beds; not-for-profit, 97% [n = 64]; teaching, 80% [n = 53]). The range in charges was substantial; for example, a screening mammogram was $40 at one hospital in Los Angeles, Calif, and $346 at one hospital in Quebec City. Charges for a screening mammogram were relatively stable between 1996 and 1997 (r=0.79; 95% confidence interval, 0.68-0.87) and unrelated to the hospital's location or charges for other services. The relative amount of variation in charges was similar for high-priced and low-priced services, similar for diagnostic and therapeutic services, and similar for the United States and Canada. CONCLUSIONS: Charges for the same hospital service vary substantially. Greater visibility might reduce some variation by bringing outliers into closer scrutiny. Patients seeking care and paying out-of-pocket could save financially by comparison shopping. PMID- 10826454 TI - The aneurysm detection and management study screening program: validation cohort and final results. Aneurysm Detection and Management Veterans Affairs Cooperative Study Investigators. AB - BACKGROUND: We previously reported the prevalence and associations of abdominal aortic aneurysm (AAA) in 73451 veterans aged 50 to 79 years who underwent ultrasound screening. OBJECTIVE: To understand the prevalence of and principal positive and negative risk factors for AAA, and to assess reproducibility of our previous findings. METHODS: In the new cohort of veterans undergoing screening, 52 745 subjects aged 50 to 79 without history of AAA underwent successful ultrasound screening for AAA, after completing a questionnaire on demographics and potential risk factors. RESULTS: We detected AAA of 4.0 cm or larger in 613 participants (1.2%; compared with 1.4% in the earlier cohort). The direction and magnitude of the important associations reported in the first cohort were confirmed. Respective odds ratios for the major associations with AAA for the second and for the combined cohorts were as follows: 1.81 and 1.71 for age (per 7 years), 0.12 and 0. 18 for female sex, 0.59 and 0.53 for black race, 1.94 and 1.94 for family history of AAA, 4.45 and 5.07 for smoking, 0.50 and 0.52 for diabetes, and 1.60 and 1.66 for atherosclerotic diseases. The excess prevalence associated with smoking accounted for 75% of all AAAs of 4.0 cm or larger in the total population of 126 196. Associations for AAA of 3.0 to 3.9 cm were similar but tended to be somewhat weaker. CONCLUSIONS: Our findings confirm our previous cohort findings. Age, smoking, family history of AAA, and atherosclerotic diseases remained the principal positive associations with AAA, and female sex, diabetes, and black race remained the principal negative associations. PMID- 10826455 TI - Women's understanding of the mammography screening debate. AB - BACKGROUND: The fractious public debate over mammography screening recommendations for women aged 40 to 49 years has received extensive attention in medical journals and in the press. OBJECTIVE: To learn how women interpret the mammography screening debate. METHODS: We mailed a survey to a random sample of American women 18 years and older, oversampling women of screening age (40-70 years). Sixty-six percent of women completed the survey (n = 503). MAIN OUTCOME MEASURES: The main outcome measures were women's reactions to the debate, their suggestion for the starting age for mammography screening, and their understanding of the source of the debate. RESULTS: Almost all women (95%) said that they had paid some attention to the recent discussion about mammography screening. Only 24% said the discussion had improved their understanding of mammography, while 50% reported being upset by the public disagreement among screening experts. Women's beliefs about mammography differed from those articulated by experts in the debate. Eighty-three percent believed that mammography had proven benefit for women aged 40 to 49 years, and 38% believed that benefit was proven for women younger than 40 years. Most women suggested that mammography screening should begin before age 40 years, while only 5% suggested a first mammogram should be performed at 50 years or older. In response to an open-ended question about why mammography has been controversial, 15% cited concerns about the potential harms of radiation and another 12% cited questions about efficacy. Nearly half (49%), however, identified costs as the major source of debate (eg, "Health maintenance organizations [HMOs] don't want to pay for mammography"). CONCLUSIONS: Most women paid attention to the recent debate about routine mammography screening for women aged 40 to 49 years, but many believed the debate was about money rather than the question of benefit. Policy makers issuing recommendations about implementation of large-scale mammography screening services need to consider how to effectively disseminate their message. PMID- 10826456 TI - Influenza and pneumococcal vaccination among Native American elders in a primary care practice. AB - BACKGROUND: More than 2 million Native Americans (ie, Native Americans and Native Alaskans) live in the United States; 60% reside in cities. This population, especially its elders, is especially susceptible to respiratory diseases; yet, adherence to guidelines for influenza and pneumococcal immunizations is unknown. OBJECTIVES: To evaluate how frequently older and high-risk adults received vaccinations for influenza and pneumococcal infection and to identify patient characteristics associated with adherence to published recommendations. METHODS: Retrospective medical record review of 550 Native American elders seen in an urban primary care practice defined using a culturally appropriate age threshold (> or =50 years) and standard criteria (> or =65 years). Univariate analyses examined demographic and clinical information by vaccination status. Logistic regressions identified factors associated with adherence to immunization guidelines. RESULTS: Among patients aged 50 years and older with any indication according to published recommendations, rates were low for influenza (31%) and pneumococcal (21%) immunizations. Likewise, few subjects at least 65 years of age had been immunized appropriately against influenza (38%) or pneumococcus (32%). Younger age and alcohol use were significantly associated with less frequent immunization; Medicare insurance, depression, and more health problems and taking more medications predicted significantly higher immunization rates. Aged 65 years or older and having cardiovascular disease or diabetes mellitus were specific indications significantly correlated with receipt of influenza and pneumococcal vaccine. CONCLUSIONS: Regardless of age or risk, inadequate vaccination rates were observed in elderly Native Americans. Our findings suggest the need to identify obstacles to immunization and to conduct prospective and elderly intervention studies in Native American populations. PMID- 10826457 TI - Placebo-associated blood pressure response and adverse effects in the treatment of hypertension: observations from a Department of Veterans Affairs Cooperative Study. AB - BACKGROUND: The use of placebo in clinical trials has been vigorously debated. Placebo control may be useful in disease states, such as stage 1 and stage 2 hypertension as defined by the Sixth Report of the Joint National Committee on Detection, Evaluation and Treatment of High Blood Pressure (JNC VI), in which response rates for placebo are high or close to response rates for effective therapies, or when established interventions have significant adverse effects. OBJECTIVE: To compare rates for the control of blood pressure and adverse effects of placebo vs active treatment in patients with stage 1 and stage 2 hypertension. METHODS: This study is a randomized controlled trial evaluating the blood pressure response and adverse effects of placebo vs 6 active treatments administered in 15 Veterans Affairs hypertension centers. The 1292 subjects of the Veterans Affairs Cooperative Study receiving single-drug therapy for hypertension were randomly allocated to receive treatment with 1 of 6 active drugs (n= 1105) or placebo (n=187). Treatment success was defined as maintaining a diastolic blood pressure of less than 95 mm Hg for at least 1 year. We compared treatment success rates for the control of blood pressure and adverse effects of placebo vs active treatment. Using the Kaplan-Meier method, we also compared rates of discontinuation from placebo vs active drug treatment over time as a result of adverse drug effects and blood pressure exceeding safety limits. RESULTS: At the end of the titration phase, 58 patients who were treated with placebo (31%) achieved a goal diastolic blood pressure lower than 90 mm Hg and 57 (30%) achieved success at 1 year. Older white patients who received placebo had a success rate of 38% vs 23% to 27% for the other age-race subgroups. The rates of discontinuation as a result of adverse drug effects were 13% for patients receiving placebo vs 12% for patients receiving active treatment (P=.40). The rates of discontinuation for blood pressure being too high were 14% for patients receiving placebo vs 7% for patients receiving active treatment (P=.01). CONCLUSIONS: Placebo control provides an important benchmark for both efficacy and adverse effects. It continues to have an appropriate place in certain therapeutic trials, particularly those involving the treatment of stage 1 and stage 2 hypertension. PMID- 10826458 TI - Superiority of lansoprazole vs ranitidine in healing nonsteroidal anti inflammatory drug-associated gastric ulcers: results of a double-blind, randomized, multicenter study. NSAID-Associated Gastric Ulcer Study Group. AB - BACKGROUND: The usefulness of nonsteroidal anti-inflammatory drugs (NSAIDs) is limited by adverse gastrointestinal tract events. OBJECTIVE: To identify the optimal antisecretory therapy for healing of gastric ulcer in patients using NSAIDs and the impact of concurrent Helicobacter pylori infection on ulcer healing. DESIGN: Prospective, double-blind, multicenter, parallel-group study. SETTING: Gastroenterology practices in ambulatory and referral center settings. PATIENTS: Three hundred fifty-three patients with an active, nonmalignant gastric ulcer at least 5 mm in diameter confirmed by endoscopy and biopsy and who continued to receive stable doses of NSAIDs. INTERVENTION: Patients were randomized to receive ranitidine hydrochloride, 150 mg twice daily, or lansoprazole, 15 mg or 30 mg once daily, for 8 weeks. MEASUREMENTS: Healing was assessed by endoscopy at 4 and 8 weeks in an intent-to-treat population. Helicobacter pylori status was assessed by histological examination. RESULTS: After 8 weeks of treatment, healing was observed in 61 (53%) of 115, 81 (69%) of 118, and 85 (73%) of 117 patients receiving ranitidine lansoprazole, 15 mg, and lansoprazole, 30 mg, respectively (P<.05 for ranitidine vs both lansoprazole doses; 95% confidence interval, 3.2-28.0 for ranitidine vs lansoprazole, 15 mg, and 7.4-31.8 for ranitidine vs lansoprazole, 30 mg). The gastric ulcer healing rates were similar between H pylori-infected and -noninfected patients, with a statistically significant increase with the use of lansoprazole vs ranitidine. CONCLUSIONS: In patients who require continuous treatment with NSAIDs, lansoprazole is superior to ranitidine for healing of NSAID-associated gastric ulcers. Healing is not delayed by the presence of H pylori infection. PMID- 10826459 TI - Effects of celecoxib and naproxen on renal function in the elderly. AB - OBJECTIVE: To compare the effects of celecoxib, a cyclooxygenase 2-specific inhibitor, with the nonspecific cyclooxygenase 1 and 2 inhibitor naproxen on renal function in 29 healthy elderly subjects in a single-blind, randomized, crossover study. METHODS: Subjects received either celecoxib, 200 mg twice daily, for 5 days followed by celecoxib, 400 mg twice daily, for the next 5 days, or they received naproxen, 500 mg twice daily, for 10 days. After a 7-day washout, subjects were crossed over to receive the other regimen. RESULTS: After the first dose, the trend was for a greater decrease in glomerular filtration rate with naproxen (-5.31 mL/min per 1.73 m2) compared with celecoxib (-0.86 mL/min per 1.73 m2). The treatment difference became statistically significant on day 6 ( 7.53 vs -1.11 mL/min per 1.73 m2 for naproxen and celecoxib, respectively; P=.004). Urinary prostaglandin E2 and 6-keto-prostaglandin F1alpha excretion was significantly reduced from baseline across the treatment interval with both celecoxib and naproxen (P< or =.04). There were no significant differences in prostaglandin excretion between these 2 agents (P> or =.07). Small, transient decreases (P<.05) in urinary sodium excretion were observed after the initiation of both celecoxib and naproxen treatment. Sodium excretion values returned to baseline by the end of the study. CONCLUSIONS: The results indicate that cyclooxygenase 2-specific inhibition in healthy elderly subjects may spare renal hemodynamic function, although the effects on sodium excretion, as well as urinary prostaglandin E2 and 6-keto-prostaglandin F1alpha excretion, appear to be similar to those of nonspecific cyclooxygenase inhibitors such as naproxen. PMID- 10826460 TI - Low birth weights contribute to high rates of early-onset chronic renal failure in the Southeastern United States. AB - BACKGROUND: The southeastern United States is a region in which rates of cardiovascular and renal diseases are excessive. Within the Southeast, South Carolina has unusually high rates of end-stage renal disease (ESRD) in young people, with more than 70% of cases attributed to hypertension and diabetes. OBJECTIVE: To determine whether the increased vulnerability to early-onset ESRD might originate through impaired renal development in utero as measured by low birth weight. METHODS: Patients who were diagnosed with renal failure and undergoing dialysis from 1991 through 1996 were identified from the ESRD registry maintained by the Southeastern Kidney Council, Raleigh, NC. Birth weights reported on birth certificates were selected for the ESRD cases and non-ESRD controls who were born in South Carolina in 1950 and later. Birth weights were compared for 1230 cases and 2460 controls who were matched for age, sex, and race. RESULTS: Low birth weight was associated with ESRD among men and women as well as blacks and whites. Among people whose birth weight was less than 2.5 kg, the odds ratio for ESRD was 1.4 (95% confidence interval, 1.1-1.8) compared with people who weighed 3 to 3.5 kg. This association was present for renal failure resulting from diabetes, hypertension, and other causes. CONCLUSIONS: Low birth weights, which reflect adverse effects on development in utero, contribute to the early onset of ESRD in South Carolina. Since low birth weight increases the risk of ESRD from multiple causes, the data suggest that an adverse environment in utero impairs kidney development and makes it more vulnerable to damage from a range of pathological processes. PMID- 10826461 TI - Medical therapy, symptoms, and the distress the cause: relation to quality of life in patients with angina pectoris and/or hypertension. AB - BACKGROUND: Adverse events during drug therapy can be assessed through measurement of 2 features: their frequency and their severity. Their severity, in turn, can be measured by assessing the distress that they cause. Our goal was to relate the magnitude of the distress induced by treatment with calcium-channel blocking agents to the change in quality of life assessed through psychosocial instruments in patients treated with calcium-channel blocking agents, either for hypertension or for angina pectoris. METHODS: Four hundred seventy-five patients with angina pectoris were randomized to double-blind treatment with PPR (physiological pattern release) verapamil hydrochloride, amlodipine besylate, amlodipineatenolol combination, or placebo. In addition, 557 hypertensive patients were randomized either to PPR verapamil or nifedipine GITS (gastrointestinal system). Both studies were double-blind. RESULTS: Significant differences in treatment of angina pectoris or hypertension, were not found between the regimens. Overall quality of life also failed to show a significant difference in either group. In both groups, however, remarkable concordance was found between the degree of distress associated with specific symptoms and a change in quality of life. An unchanged, stable symptom distress was associated with a significant improvement in the quality of life of about 0.1 SD. Improvement or erosion of symptom distress represented by 1 step was associated with a 0.1- to 0.2-SD change. The extreme change in symptom distress was associated with a substantially larger change in global quality of life. CONCLUSIONS: The magnitude of symptom distress or relief associated with symptoms in 2 patient populations correlated strongly with a shift in quality of life. The assessment of distress associated with symptoms provides valuable additional information on drug therapy. PMID- 10826462 TI - Comparison of central nervous system adverse effects of amantadine and rimantadine used as sequential prophylaxis of influenza A in elderly nursing home patients. AB - BACKGROUND: Amantadine hydrochloride and rimantadine hydrochloride are recommended by the Centers for Disease Control and Prevention for prophylaxis of influenza A. While data suggest that rimantadine is better tolerated, there are no data examining the rate of adverse reactions in elderly patients who receive amantadine vs rimantadine. Our objective was to assess the adverse reaction rate in elderly nursing home patients receiving sequential amantadine and rimantadine for influenza A prophylaxis. METHODS: Data were collected in 156 nursing home patients (70% women; mean+/-SD age, 83.7+/-10.1 years) in a single care setting who received sequential therapy with amantadine and rimantadine during the 1997 1998 influenza season. Patients were assessed for central nervous system adverse effects and therapy discontinuation occurring with each agent. RESULTS: Twenty nine (18.6%) of the 156 patients experienced an adverse effect when receiving amantadine compared with 3 patients (1.9%) when rimantadine was given (P<.01). Drug use was discontinued due to adverse events in 17.3% (n = 27) of the amantadine courses and 1.9% (n=3) of the rimantadine courses (P<.001). Confusion was the most frequently observed adverse event (amantadine, 10.6%; rimantadine, 0.6%; P<.001). Multivariate logistic regression analysis showed that significant risk factors for central nervous system adverse events included male sex (odds ratio, 3.65), reduced calculated creatinine clearance (odds ratio, 1.78), and use of amantadine (odds ratio, 12.73). CONCLUSIONS: Amantadine use was associated with a significantly higher incidence of central nervous system adverse events than rimantadine use in this elderly population receiving influenza prophylaxis. In addition, the discontinuation rate of amantadine was significantly higher than that with rimantadine. PMID- 10826463 TI - Increased prevalence of celiac disease in patients with dyspepsia. AB - BACKGROUND: Although 30% to 40% of patients with celiac disease (CD) (which affects 1 in 200 individuals) have dyspeptic symptoms, there is a lack o data concerning the prevalence of CD in patients with dyspepsia. METHODS: In this prospective series, we enrolled all consecutive outpatients undergoing endoscopy of the upper gastrointestinal tract for dyspepsia at our centers between January and June 1998. The exclusion criteria were age younger than 12 years, workup or follow-up of an already known disease of the gastrointestinal tract, suspected CD, malabsorption, and/or iron-deficiency anemia. RESULTS: Of the 3019 patients who were evaluated, 517 (17%) were eligible for the study. Endoscopic findings suggested CD in 5 cases. Celiac disease was histologically diagnosed in 6 patients (5 women and 1 man; mean age, 31.3 years; age range, 20-46 years), 3 of whom had a normal endoscopic pattern and 3 of whom had an endoscopic pattern that was consistent with CD. In the patients with histologically diagnosed CD, antiendomysium antibody positivity supported the diagnosis. The relative risk for CD was 2.32 (95% confidence interval, 1.06-5.07) in comparison with the general population and higher among females (3.22; 95% confidence interval, 1.37-7.56). CONCLUSIONS: The present results indicate that the prevalence of CD in patients with dyspepsia is twice that of the general population. Thus, serological screening for CD should be considered in the early workup of these patients to allow diagnosis and treatment of an eminently treatable disease. PMID- 10826464 TI - Do depression symptoms predict early hypertension incidence in young adults in the CARDIA study? Coronary Artery Risk Development in Young Adults. AB - BACKGROUND: Hypertension has been linked to several psychological factors, including depression, but the relation between hypertension incidence and depressive symptoms has not been adequately examined. OBJECTIVE: To determine if depressive symptoms independently predict hypertension incidence. DESIGN AND SETTING: A prospective, multicenter, epidemiological cohort of young adults (aged 23-35 years at study entry) from the general community without hypertension followed up for 5 years. SUBJECTS: A sample of 3343 adults from 4 urban areas stratified for race (black and white) from the CARDIA (Coronary Artery Risk Development in Young Adults) study. MAIN OUTCOME MEASURE: Hypertension incidence, which was defined as blood pressure higher than 160/95 mm Hg (assessed on a single occasion) or the use of prescribed antihypertensive medication. RESULTS: Participants with high scores (> or = 16) on the Center for Epidemiological Studies Depression (CES-D) Scale were at significant risk for hypertension incidence compared with those with low CES-D scores (< or =7; odds ratio, 2.10; 95% confidence interval, 1.22-3.61) after adjustment for other hypertension risk factors (eg, age, resting systolic blood pressure at the 5-year examination, physical activity, daily alcohol use, parental history of hypertension, education, presence of diabetes mellitus or heart disease, sex, and race) in fixed logistic models. Those with intermediate depressive symptoms (CES-D scores 8-15) were also at significant risk (adjusted odds ratio, 1.78; 95% confidence interval, 1.06-2.98). These associations were significant in blacks alone but were not found in whites, who had a lower hypertension incidence (29 [2%] of 1806) than blacks (89 [6%] of 1537). CONCLUSIONS: Depressive symptoms were predictive of later hypertension incidence in young adults, and young blacks with depressive symptoms were at high risk of developing hypertension. PMID- 10826465 TI - Older person's preferences for home vs hospital care in the treatment of acute illness. AB - BACKGROUND: Although the home is expanding as a potential site for acute illness treatment, little is known about patients' preferences for home vs the hospital. OBJECTIVE: To determine older persons' preferences for home or hospital as a treatment site for acute illness and factors associated with preference. METHODS: Two hundred forty-six community-dwelling persons aged 65 years or older hospitalized with congestive heart failure, chronic obstructive pulmonary disease, or pneumonia were identified in 2 urban teaching hospitals and received telephone interviews 2 months after hospitalization. They were asked their preference for home or hospital treatment, given the availability of equivalent therapies and outcomes at the 2 sites and a nursing visit and several hours of home health aide assistance daily in the home. They were also asked about changes in preference with changes in the description of the outcome or the availability of services. RESULTS: If home and hospital offered equivalent outcomes, 46% of the sample preferred treatment at home. Preferences were heavily dependent on the outcome of the illness, physician opinion about the best site of care, and the provision of house calls. Higher education, white race, living with a spouse, being deeply religious, and having 2 or more dependencies in activities of daily living were associated with a preference for home treatment. CONCLUSIONS: Under conditions of equivalent outcome, preferences for treatment site are almost equally divided between home and hospital. Explicit elucidation of preferences is necessary if patients' preferences are to play a meaningful role in decision making about site of care. PMID- 10826466 TI - White-coat hypertension and carotid artery atherosclerosis: a matching study. AB - BACKGROUND: Blood pressure (BP) measurements obtained in the clinic have long served as the basis for determining risk of hypertensive vascular disease, yet many patients with high BP in the physician's office are normotensive elsewhere. It remains unclear whether such patients with "white coat" hypertension elude the risk of atherosclerosis. METHODS: Community residents 40 to 70 years of age and not receiving any cardiovascular medications were recruited to participate in a study of cardiovascular risk factors. On the basis of clinic and daytime ambulatory BP and a threshold criterion of 140/90 mm Hg, subjects were classified as having persistent hypertension, white-coat hypertension, or persistent normotension. One-to-one matching was conducted in male participants on the basis of race and BP. Subjects with persistent hypertension and white-coat hypertension were matched on clinic BP, and those with white-coat hypertension and normotension were matched on daytime ambulatory BP. RESULTS: The 3 matched groups of men (n=40 in each group) were similar in age, smoking status, and fasting glucose and lipid levels. Compared with the normotensive subjects, subjects with either persistent or white-coat hypertension had greater mean body mass index, waist-hip ratio, and fasting insulin concentration. On the basis of standardized duplex ultrasound examination of the carotid arteries, mean maximal intimal medial thickness and plaque index in subjects with white-coat hypertension were greater than among normotensive subjects and equal to that of the subjects with persistent hypertension. CONCLUSION: When compared with unmedicated individuals with comparable elevations in clinic BP, individuals with white-coat hypertension appear not to be protected from the atherosclerotic sequelae of hypertension. PMID- 10826467 TI - Benefits of screening for latent Mycobacterium tuberculosis infection. AB - BACKGROUND: The benefits of screening for latent Mycobacterium tuberculosis infection are unknown for most people, because screening has not been studied in clinical trials and preventive therapy has not been tested in all risk groups for whom it is recommended. METHOD: A MEDLINE search was performed to determine tuberculosis risk. A Markov model was used to analyze tuberculin skin test screening and preventive therapy for 3-year-old and 30-year-old persons with positive test results. Outcome measures were lifetime and 10-year tuberculosis risk, including spread to others, life expectancy extension, and number needed to screen and number needed to treat to prevent 1 case and 1 death during 10 years. RESULTS: The benefits of screening and preventive therapy outweigh the risks for all groups tested, although the benefits range from large to small. The number needed to screen to prevent 1 case is 10 to 6888, and the number needed to treat is 2 to 179. Persons with human immunodeficiency virus infection, intravenous drug abuse, or end-stage renal disease treated with transplantation and children exposed to high-risk adults have the highest tuberculosis rates and the lowest number needed to screen and number needed to treat to prevent cases and deaths. The range of risks found in the literature for some risk groups, such as persons with silicosis, leukemia or lymphoma, end-stage renal disease treated with dialysis, or prolonged corticosteroid therapy, is wide and, as a result, the benefits of screening are uncertain. CONCLUSIONS: The benefits of screening and preventive therapy vary widely, although the benefits outweigh the risks for all risk groups. The benefits are large for some risk groups and uncertain for others. PMID- 10826468 TI - Suicidal ideation and suicide attempts in general medical illnesses. AB - BACKGROUND: This study examines the association between the presence of a general medical illness and suicidality in a representative sample of US young adults. METHODS: Between 1988 and 1994, 7589 individuals aged 17 to 39 years were administered the Diagnostic Interview Schedule as part of a national probability survey. The survey collected information about lifetime suicidal ideation and suicide attempts, a checklist of common general medical conditions, and data on major depression, alcohol use, and demographic characteristics. RESULTS: Whereas 16.3% of respondents described suicidal ideation at some point in their lives, 25.2% of individuals with a general medical condition, and 35.0% of those with 2 or more medical illnesses reported life-time suicidal ideation. Similarly, whereas 5.5% of respondents had made a suicide attempt, 8.9% of those with a general medical illness and 16.2% of those with 2 or more medical conditions had attempted suicide. In models controlling for major depression, depressive symptoms, alcohol use, and demographic characteristics, presence of a general medical condition predicted a 1.3 times increase in likelihood of suicidal ideation; more specifically, pulmonary diseases (asthma, bronchitis) were associated with a two-thirds increase in the odds of lifetime suicidal ideation. Cancer and asthma were each associated with a more than 4-fold increase in the likelihood of a suicide attempt. CONCLUSIONS: A significant association was found between medical conditions and suicidality that persisted after adjusting for depressive illness and alcohol use. The findings support the need to screen for suicidality in general medical settings, over and above use of general depression instruments. PMID- 10826469 TI - Pulmonary embolism as a cause of cardiac arrest: presentation and outcome. AB - BACKGROUND: Pulmonary embolism (PE) is a possible noncardiac cause of cardiac arrest. Mortality is very high, and often diagnosis is established only by autopsy. METHODS: In a retrospective study, we analyzed clinical presentation, diagnosis, therapy, and outcome of patients with cardiac arrest after PE admitted to the emergency department of an urban tertiary care hospital. RESULTS: Within 8 years, PE was found as the cause in 60 (4.8%) of 1246 cardiac arrest victims. The initial rhythm diagnosis was pulseless electrical activity in 38 (63%), asystole in 19 (32%), and ventricular fibrillation in 3 (5%) of the patients. Pronounced metabolic acidosis (median pH, 6.95, and lactate level, 16 mmol/L) was found in most patients. In 18 patients (30%), the diagnosis of PE was established only postmortem. In 42 (70%) it was diagnosed clinically, in 24 of them the diagnosis of PE was confirmed by echocardiography. In 21 patients, 100 mg of recombinant tissue-type plasminogen activator was administered as thrombolytic treatment, and 2 (10%) of these patients survived to hospital discharge. Comparison of patients of the thrombolysis group (n = 21) with those of the nonthrombolysis group (n = 21) showed a significantly higher rate of return of spontaneous circulation (81% vs 43%) in the thrombolysis group (P=.03). CONCLUSIONS: Mortality related to cardiac arrest caused by PE is high. Echocardiography is supportive in determining PE as the cause of cardiac arrest. In view of the poor prognosis, thrombolysis should be attempted to achieve return of spontaneous circulation and probably better outcome. PMID- 10826470 TI - Small-vessel vasculitis in granulomatous giant cell arteritis. PMID- 10826471 TI - Human immunodeficiency virus testing and behavior change. PMID- 10826472 TI - Identifying high-risk surgical patients by poor self-reported exercise tolerance. PMID- 10826473 TI - Overestimation of the number of individuals with hypertension who are eligible for treatment according to the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure. PMID- 10826474 TI - Thyroid nodules and carcinoma in Graves disease. PMID- 10826475 TI - Hypernatremia in hospitalized patients: a sequel of inadvertent fluid administration. PMID- 10826476 TI - Genetic characterization of incident HIV type 1 subtype E and B strains from a prospective cohort of injecting drug users in Bangkok, Thailand. AB - We obtained specimens from 128 HIV-1 seroconverters identified from 1995 through 1998 in a prospective cohort study of 1,209 HIV-negative injecting drug users (IDUs) in Bangkok, Thailand. Epidemiologic data indicated that parenteral transmission accounted for nearly all infections. HIV-1 DNA from the C2-V4 env region was sequenced, and phylogenetic analyses determined that 102 (79.7%) of the specimens were subtype E and 26 (20.3%) subtype B strains. All subtype B strains clustered with strains often referred to in previous studies as Thai B or B'. The interstrain nucleotide distance (C2-V4) within subtype E strains was low (mean, 6.8%), and pairwise comparisons with a prototype subtype E strain, CM244, showed limited divergence (mean, 5.6%). The subtype B stains showed greater interstrain divergence (mean, 9.2%) and were significantly divergent from the prototype B strain HIV-MN (mean, 13.0%; p < 0.0001). The subtype E strains had significantly lower mean V3 loop charge than did subtype B strains (p = 0.017) and, on the basis of analysis of amino acid sequences, were predicted to be predominantly (91%) non-syncytium-inducing (NSI), chemokine coreceptor CCR5-using (CCR5+) viruses. The subtype B strains had a higher mean V3 loop charge, and a smaller proportion (23%) were predicted to be NSI/CCR5+ viruses. This study demonstrates that most incident HIV1 infections among Bangkok IDUs are due to subtype E viruses, with a narrow spectrum of genetic diversity. The characterization of incident HIV-1 strains from 1995 to 1998 will provide important baseline information for comparison with any breakthrough infections that occur among IDUs in Bangkok who are participating in an HIV-1 vaccine efficacy trial initiated in 1999. PMID- 10826477 TI - Deletion of the p16INK4A gene in ex vivo acute adult T cell lymphoma/leukemia cells and methylation of the p16INK4A promoter in HTLV type I-infected T cell lines. AB - The stoichiometry of the p16INK4A and p15INK4B proteins bound to the cyclin D CDK4/6 complex regulates the entry of cells into the G1 phase of the cell cycle. Thus, their level of expression is essential in maintaining regulated cell growth. In several tumors, deletion of these genes has been reported and, more recently, promoter methylation has been suggested as an alternative mechanism to decrease the expression of these cell cycle inhibitor proteins. Here, we studied the methylation status and the integrity of the p16INK4A and p15INK4B genes in 8 chronically HTLV-I-infected T cell lines and in ex vivo cells from 14 ATLL patients. Deletion of the locus carrying both genes was not found in the HTLV-I infected T cell lines but was found in seven of eight acute ATLL cases and in none of the PBMCs from the chronic cases or the affected lymph nodes of the lymphoma type. In contrast, partial or complete methylation of one or both genes was found only in chronically HTLV-I T cells. Thus, HTLV-I infection targets the p16INK4A and p15INK4B loci both in vitro and in vivo, although the mechanisms may differ. PMID- 10826478 TI - HTLV-I/HTLV-II coinfection in an AIDS patient from Sao Paulo, Brazil. AB - A serological survey for HTLV infection identified an AIDS patient with HTLV I/HTLV-II dual seroreactivity. Two further sequential blood samples were collected (samples A and B) for PCR, restriction fragment length polymorphism (RFLP), and sequence analyses of HTLV-I and HTLV-II strains. PCR analyses confirmed dual infection in both samples. Restriction digests of the env region amplified from sample A showed the presence of an HTLV-IIa subtype; the HTLV-II provirus was found to be defective in the pol and env regions in the second sample from this patient. RFLP analysis of the HTLV-II LTR region of both samples confirmed this finding and identified an a5/bzl restriction type. Nucleotide sequence analyses revealed full homology in the HTLV-I env and LTR regions and in the HTLV-II LTR region between the two samples. These findings document the first case of an HTLV-I/HTLV-II coinfection that was fully confirmed and characterized by means of molecular analyses. PMID- 10826479 TI - Rapid identification of all known retroviral reverse transcriptase sequences with a novel versatile detection assay. AB - We have developed a highly sensitive, universal assay that allows detection as well as identification of all known retroviral reverse transcriptase (RT)-related nucleic acids in a biological sample by a single two-step experiment. The assay combines polymerase chain reaction (PCR) and reverse dot-blot hybridization (RDBH), using an array of immobilized synthetic retrovirus-specific oligonucleotides and two sets of mixed oligo primers (MOPs). These primers were derived from highly conserved motifs found in all known reverse transcriptase genes. The PCR/RDBH assay was used for qualitative analyses of human endogenous retrovirus (HERV) transcription in peripheral blood mononuclear cells (PBMCs) and in particles released by the human mammary carcinoma-derived cell line T47D. Sensitivity was further demonstrated by detection of down to 10 copies of pig endogenous retrovirus (PERV) DNA in human cDNA samples. Therefore, this assay is particularly useful for the identification of retroviral sequences in xenografts as well as in recipients of xenografted tissues and organs. Moreover, it is a valuable tool to detect retroviral transcripts and particles in cell cultures used for production of therapeutic polypeptides. The assay is further suitable for monitoring vector preparation used in human gene therapy to exclude transfer of copackaged endogenous retroviruses into target cells. PMID- 10826480 TI - Chromosomal distribution and coding capacity of the human endogenous retrovirus HERV-W family. AB - Some genomic elements of the multicopy HERV-W endogenous retroviral family have been previously identified in databases. One of them, located on chromosome 7, contains a single complete open reading frame (ORF) putatively encoding an envelope protein. We have experimentally investigated the genomic complexity and coding capacity of the HERV-W family. The human haploid genome contains at least 70, 100, and 30 HERV-W-related gag, pro, and env regions, respectively, widely and heterogeneously dispersed among chromosomes. Using in vitro transcription translation procedures, three putative HERV-W gag, pro, and env ORFs were detected on chromosomes 3, 6, and 7, respectively, and their sequences analyzed. A 363 amino acid gag ORF containing matrix and carboxy-terminal truncated capsid domains encoded a putative 45-kDa protein. No gag-pro ORF was found, but a pro sequence containing a DTG active site was detected. Finally, the previously described 538 amino acid HERV-W env ORF, located on chromosome 7, was shown to be unique and encoded a putative 80-kDa glycosylated protein. Proteins of molecular mass identical to the one obtained by an in vitro transcription-translation procedure were detected in human placenta, using anti HERV-W Gag- and Env specific antibodies. The absence of an HERV-W replication-competent provirus versus the existence of HERV-W-related Gag and Env proteins in healthy human placenta is discussed with respect to particle formation, physiology, and pathology. PMID- 10826481 TI - Fine definition of a conserved CCR5-binding region on the human immunodeficiency virus type 1 glycoprotein 120. AB - A previous study implicated a conserved surface of the human immunodeficiency virus (HIV-1) gp120 exterior envelope glycoprotein in binding the CCR5 viral coreceptor (Rizzuto C, Wyatt R, Hernandez-Ramos N, Sun Y, Kwong PD, Hendrickson WA, and Sodroski J: Science 1998;280:1949-1953). Additional mutagenesis indicates that important residues in this region for CCR5 binding are Ile-420, Lys-421, Gln 422, Pro-438, and Gly-441. These highly conserved residues are located on two strands that connect the gp120 bridging sheet and outer domain, suggesting a mechanism whereby interdomain conformational shifts induced by CD4 binding could facilitate CCR5 binding. PMID- 10826482 TI - In vitro and in vivo responses to interleukin 12 are maintained until the late SIV infection stage but lost during AIDS. AB - The in vitro proliferative responses of macaque peripheral blood mononuclear cells (PBMCs) to IL-12 appeared similar before and early after SIV infection, whereas macaque PBMCs sampled during symptomatic stages of SIV infection showed markedly decreased responses. IL-12 was administered to SIVmac239-infected rhesus macaques either during the asymptomatic or the AIDS stage of infection in efforts to evaluate the effect of this cytokine on immune responses, viral loads, and hematopoietic functions in vivo. IFN-gamma secretion levels induced during the asymptomatic or early symptomatic phase were similar to preinfection induced levels, whereas in later AIDS stages this response was lost. The constitutive levels of other measured cytokines were not affected by IL-12 administration in vivo. The frequency and activity of circulating NK cells were markedly enhanced at early stages but not at symptomatic stages of SIV infection. pCTL frequencies were enhanced at early symptomatic stages but not at late AIDS stages. Despite its immunomodulatory effect, IL-12 did not seem to exacerbate or inhibit the replication of SIV in vivo, or the frequency of circulating infected lymphocytes. IL-12 administration was associated with a significant yet subclinical and transient decrease in hematocrit and hemoglobin levels without evidence of hemolysis, hemodilution, or reduction in the frequency of colony-forming unit potential of bone marrow CD34+ cells. This phenomenon may be explained by a functional inhibition of differentiation rather than an altered generation of bone marrow precursors. Thus, these results suggest that IL-12 may benefit HIV-1 infected patients only as long as their immune system retains its capability to respond to cytokine stimulation. PMID- 10826483 TI - Higher western blot immunoreactivity of glycoprotein 120 from R5 HIV type 1 isolates compared with X4 and X4R5 isolates. AB - The envelope glycoprotein of human immunodeficiency virus 1 (HIV-1) plays important roles in viral life cycle and pathogenesis. Understanding the immune responses the protein elicits during the course of a viral infection in patients is important in designing an effective vaccine candidate against the virus or for better diagnosis of the disease. In this study, we report that gp120 of R5 isolates have higher Western blot (WB) immunoreactivity to antibodies elicited against the protein in virus-infected human patients compared with that of X4 and X4R5 isolates. Analyses of WB immunoreactivity of chimeric gp120s constructed between R5 (AD8) and X4R5 (DH12) HIV-1 isolates indicate that there are complex tertiary interdomain interactions even after a complete denaturation of the protein. Our data suggest that the determinant(s) responsible for the high WB immunoreactivity might be present in all gp120s, but are accessible to antibodies only for R5 gp120s in the WB assay. The V1/V2 and/or V3 regions of X4 and X4R5 gp120s likely interfere with either the formation or surface exposure of the WB immunoreactive determinant. Supplementing HIV-1 WB diagnosis kits with purified R5 gp120 could improve their sensitivity and facilitate earlier diagnosis of virus infection. PMID- 10826484 TI - Nef modulation of HIV type 1 gene expression and cytopathicity in tissues of HIV transgenic mice. AB - Transgenic mice bearing HIV-1 proviral DNA deleted in the gag/pol region (HIVd1443 mice) model a chronic, nonproductive form of viral gene expression in various cell types including macrophages. They display a disease phenotype that includes HIV-associated nephropathy (HIVAN), congenital cataracts, papillomatosis, and growth failure. The role of HIV-1 Nef in viral gene regulation and the development of disease was explored in mice bearing an isogenic HIV transgene in which nef was mutated by frameshift mutation. Like its Nef+ counterpart, HIVd1443[Nef-] mice expressed HIV gene products in the skin, muscle, kidney, and peritoneal macrophages. While these mice did not develop cataracts, papillomatous skin lesions, or display any apparent growth defect, they did develop HIVAN. Nef expression was introduced to HIVd1443[Nef-] mice through breeding to mice bearing an HIV LTR-linked nef transgene. Nef complemented HIVd1443[Nef-] mice had reduced levels of viral gene products in the muscle and kidney. In contrast, HIV gene expression in the skin of these mice remained high and papillomatous lesions emerged that were more severe than those on wild-type HIVd1443 mice. Still, Nef had a negative effect on LPS-induced viral gene expression in visibly normal skin. In comparisons of peritoneal macrophages, viral RNA expression was significantly reduced in resident macrophages of Nef+ mice. HIV inflammatory macrophages expressed viral genes and displayed an altered FACS profile. In particular, Nef+ populations were marked by an increased proportion of F4/80med/Mac-1-cells as well as fewer Mac-1 cells and reduced F4/80 staining. This HIV proviral transgenic model has demonstrated the capacity of HIV 1 Nef to contribute to HIV cytopathicity by altering cellular maturation and viral gene expression in vivo. PMID- 10826485 TI - Effect of PMPA and PMEA on the kinetics of viral load in simian immunodeficiency virus-infected macaques. AB - In this study we compared the effect of postexposure treatment of the acyclic nucleoside analogs 9-(2-phosphonylmethoxyethyl)-adenine (PMEA) and 9-(2 phosphonylmethoxypropyl)-adenine (PMPA) on the kinetics of viral load in the blood and lymph nodes of rhesus macaques chronically infected with SIVmac251 for 18 weeks. Two of the four macaques treated with PMPA (20 mg/kg per day) for 28 consecutive days had demonstrable reductions in viral loads of 1.5 and 3 logs. Three of four macaques given the same dosing regimen of PMEA had viral load reductions ranging from 1.25 to 2.8 logs. Furthermore, treatment with either drug caused a reduction in virus burden in the lymph nodes by 2 weeks posttreatment. However, in both PMEA- and PMPA-treated animals, viral loads rebounded to day of treatment levels by 2 weeks after termination of treatment. The extent to which viral load was suppressed was similar for both drugs. In contrast, viral loads in three of four mock-treated animals remained persistently high throughout the study. This study has demonstrated that postexposure treatment with these acyclic nucleoside analogs could modulate the kinetics of viral load reduction in some animals. PMID- 10826486 TI - Diversity of envelope glycoprotein from human immunodeficiency virus type 1 of recent seroconverters in Thailand. AB - The envelope-coding sequence of human immunodeficiency virus type 1 (HIV-1) was determined for 11 Thai seroconverters between 1995 and 1996. On the basis of the env sequences, all subjects were infected with HIV subtype E. Compared with the interpatient protein diversity among HIV-1 Thai reference sequences from 1990 to 1992 (4.4%), the diversity among the 1995-1996 seroconverters was approximately double (9.5%). The tetrapeptide tip of the V3 loop was invariant for 10 of the 11 seroconverters, and identical to that observed in sequences derived from the 1990 1992 group. However, in the V3 region, sequences from 2 of the 11 subjects demonstrated more than 5 amino acid changes relative to the reference strains. This may represent the "aging" of the HIV epidemic seen in other endemic regions. These findings may have substantial implications for vaccine development and evaluation for both HIV antibody and cytotoxic T lymphocyte repertoire recognition. PMID- 10826487 TI - Analysis of HIV type 1 protease and reverse transcriptase in antiretroviral drug naive Ugandan adults. AB - We analyzed plasma HIV-1 from 27 antiretroviral drug-naive Ugandan adults. Previous subtype analysis of env and gag sequences from these samples identified subtypes A, C, D, and recombinant HIV-1. Sequences of HIV-1 protease and reverse transcriptase (RT) were obtained with a commercial HIV-1 genotyping system. Subtypes based on protease sequences differed from gag subtypes for 5 of 27 samples, demonstrating a high rate of recombination between the gag and pol regions. Protease and RT sequences were analyzed for the presence of amino acid polymorphisms at positions that are sites of previously characterized drug resistance mutations. At those sites, frequent polymorphisms were detected at positions 36 and 69 in protease and positions 179, 211, and 214 in RT. Subtype specific amino acid motifs were identified in protease. Most of the subtype A sequences had the amino acids DKKM at positions 35, 57, 69, and 89, whereas most subtype D sequences had the amino acids ERHL at those positions. Detection of those polymorphisms may provide a useful approach for rapid identification of subtype A and D isolates in Uganda. This analysis significantly increases the number of Ugandan protease and RT sequences characterized to date and demonstrates successful use of a commercial HIV-1 genotyping system for analysis of diverse non-B HIV-1 subtypes. PMID- 10826488 TI - Genetic characterization and phylogenetic analysis of HIV-1 subtype C from Uganda. AB - To better understand the emergence of subtype C and its potential impact on vaccine efforts in Uganda, we have characterized subtype C sequences from Uganda (n = 13), Zimbabwe (n = 11), Mozambique (n = 5), South Africa (n = 4), and India (n = 3). Phylogenetic analysis of subtype C sequences in the env gp41 gene region revealed multiple subclusters within subtype C. Further, while most Ugandan specimen subclustered together, other subclusters did not reflect a clear geographic location. The nucleotide divergence within the Ugandan subset was 8.2% (6.1-9.8%) compared with 9.5% (2.5-15%) for the other subtype C gp41 sequences. The protein sequence alignment revealed marked sequence conservation of major immunodominant epitopes within the gp41 region. PMID- 10826489 TI - Mutations in the reverse transcriptase gene of HIV type 1 from subjects after stavudine-didanosine dual therapy. PMID- 10826490 TI - Localization of ribophorin II to the endoplasmic reticulum involves both its transmembrane and cytoplasmic domains. AB - Proteins that are concentrated in specific compartments of the endomembrane system in order to exert their organelle-specific function must possess specific localization signals that prevent their transport to distal regions of the exocytic pathway. Some resident proteins of the endoplasmic reticulum (ER) that are known to escape with low efficiency from this organelle to a post ER compartment are recognized by a recycling receptor and brought back to their site of residence. Other ER proteins, however, appear to be retained in the ER by mechanisms that operate in the organelle itself. The mammalian oligosaccharyltransferase (OST) is a protein complex that effects the cotranslational N-glycosylation of newly synthesized polypeptides, and is composed of at least four rough ER-specific membrane proteins: ribophorins I and II (RI and RII), OST48, and Dadl. The mechanism(s) by which the subunits of this complex are retained in the ER are not well understood. In an effort to identify the domains within RII responsible for its ER localization we have studied the fate of chimeric proteins in which one or more RII domains were replaced by the corresponding ones of the Tac antigen, the latter being a well characterized plasma membrane protein that lacks intrinsic ER retention signals and serves to provide a neutral framework for the identification of retention signals in other proteins. We found that the luminal domain of RII by itself does not contain retention information, while the cytoplasmic and transmembrane domains contain independent ER localization signals. We also show that the retention function of the transmembrane domain is strengthened by the presence of a flanking luminal region consisting of 15 amino acids. PMID- 10826491 TI - The membrane transport factor p115 recycles only between homologous compartments in intact heterokaryons. AB - Cytosolic proteins that participate in membrane traffic are assumed to be recruited from the cytosol onto specific membrane sites where they perform their function, and then released into cytosol before rebinding to catalyze another round of transport. To examine whether the ER to Golgi transport factor p115 recycles through release into a cytosolic pool, we formed heterokaryons between rat NRK and simian COS-7 cells and examined the dynamics of rat p115 transfer from the rat to the simian portion of the heterokaryon. The heterokaryons shared a common cytosolic pool, as shown by the efficient relocation of a cytosolic green fluorescent protein (GFP) from the COS-7 to the NRK part of the heterokaryon. Unexpectedly, even 24 h after cell fusion, rat p115 did not redistribute to the COS-7 part of the heterokaryon. This was not due to the inability of the rat p115 to associate with simian membranes since rat p115 expressed in COS-7 cells was efficiently targeted to and associated with simian Golgi complex. Furthermore, rat p115 associated with heterologous simian membranes after the NRK and COS-7 Golgi fused into a single chimeric structure. Our results indicate that p115 is not freely diffusible in intact cells and might remain tethered to membranes throughout its life cycle. These findings suggest that p115, and perhaps other cytosolic proteins involved in membrane traffic, recycle not by being released into cytosol, but in association with recycling membranes. PMID- 10826492 TI - Interaction of casein kinase 1 delta (CK1delta) with post-Golgi structures, microtubules and the spindle apparatus. AB - Members of the casein kinase 1 family of serine/threonine kinases are highly conserved from yeast to mammals and seem to play an important role in vesicular trafficking, DNA repair, cell cycle progression and cytokinesis. We here report that in interphase cells of various mammalian species casein kinase 1 delta (CK1delta) specifically interacts with the trans Golgi network and cytoplasmic, granular particles that associate with microtubules. Furthermore, at mitosis CK1delta is recruited to the spindle apparatus and the centrosomes in cells, which have been exposed to DNA-damaging agents like etoposide or gammairradiation. In addition, determination of the affinity of CK1delta to different tubulin isoforms in immunoprecipitation-Western analysis revealed a dramatically enhanced complex formation between CK1delta and tubulins from mitotic extracts after introducing DNA damage. The high affinity of CK1delta to the spindle apparatus in DNA-damaged cells and its ability to phosphorylate several microtubule-associated proteins points to a regulatory role of CK1delta at mitosis. PMID- 10826493 TI - GSK3beta-mediated phosphorylation of the microtubule-associated protein 2C (MAP2C) prevents microtubule bundling. AB - A major determinant of neuronal morphology is the cytoskeleton. And one of the main regulatory mechanisms of cytoskeletal proteins is the modification of their phosphorylation state via changes in the relative activities of protein kinases and phosphatases in neurons. In particular, the microtubule-associated protein 2 (MAP2) family of proteins are abundant cytoskeletal components predominantly expressed in neurons and have been found to be substrates for most of protein kinases and phosphatases present in neurons, including glycogen-synthase kinase 3 (GSK3). It has been suggested that changes in GSK3-mediated MAP phosphorylation may modify MT stability and could control neuronal development. We have previously shown that MAP2 is phosphorylated in vitro and in situ by GSK3 at Thr1620 and Thr1623, located in the proline-rich region of MAP2 and recognized by antibody 305. However, the function of the phosphorylation of this site of MAP2 is still unknown. In this study, non-neuronal COS-1 cells have been co transfected with cDNAs encoding MAP2C and either wild type or mutated GSK3beta to analyze possible effects on microtubule stability and on the association of MAP2 with microtubules. We have found that GSK3beta phosphorylates MAP2C in co transfected cells. Moreover, this phosphorylation is inhibited by the specific GSK3 inhibitor lithium chloride. Additionally, the formation of microtubule bundles, which is observed after transfection with MAP2C, was decreased when MAP2C was co-transfected with GSK3beta wild type. Microtubule bundles were not observed in cells expressing MAP2C phosphorylated at the site recognized by antibody 305. The absence of microtubule bundles was reverted after treatment of MAP2C/GSK3beta wild type transfected cells with lithium chloride. Highly phosphorylated MAP2C species, which were phosphorylated at the site recognized by antibody 305, appeared in cells co-transfected with MAP2C and GSK3beta wild type. Interestingly, these MAP2C species were enriched in cytoskeleton-unbound protein preparations. These data suggests that GSK3-mediated phosphorylation of MAP2 may modify its binding to microtubules and regulate microtubule stability. PMID- 10826494 TI - Perinuclear localization of the protein-tyrosine phosphatase SHP-1 and inhibition of epidermal growth factor-stimulated STAT1/3 activation in A431 cells. AB - The SH2 domain protein-tyrosine phosphatase SHP-1 has been shown earlier to bind to the epidermal growth factor receptor and to have the capacity for receptor dephosphorylation. New bi- and tricistronic expression vectors (pNRTIS-21 and pNRTIS-33, respectively) based on the tetracycline system were constructed and employed to generate stable cell lines with inducible expression of SHP-1. Inducible overexpression of SHP-1 in A431 cells led to attenuation of epidermal growth factor (EGF) receptor autophosphorylation and of EGF-induced DNA binding of 'signal transducers and activators of transcription' (STAT) 1 and 3. SHP-1 was localized in the cytoplasm with an enrichment in the perinuclear compartment. Association of SHP-1 with perinuclear structures may form the basis for a partial cofractionation with nuclei observed in different types of transfected cells and also with endogenous SHP-1 in U-937 cells. Treatment of SHP-1-overexpressing A431 cells or of HaCaT human keratinocytes expressing SHP-1 endogenously with the Ca2+ ionophore A23187 resulted in partial nuclear accumulation of SHP-1. Thus, SHP-1 may interact with substrates or regulatory proteins in perinuclear or nuclear structures. PMID- 10826495 TI - Dynamic connexin43 expression and gap junctional communication during endoderm differentiation of F9 embryonal carcinoma cells. AB - Gap junctional communication permits the direct intercellular exchange of small molecules and ions. In vertebrates, gap junctions are formed by the conjunction of two connexons, each consisting of a hexamer of connexin proteins, and are either established or degraded depending on the nature of the tissue formed. Gap junction function has been implicated in both directing developmental cell fate decisions and in tissue homeostasis/metabolite exchange. In mouse development, formation of the extra embryonal parietal endoderm from visceral endoderm is the first epithelial-mesenchyme transition to occur. This transition can be mimicked in vitro, by F9 embryonal carcinoma (EC) cells treated with retinoic acid, to form (epithelial) primitive or visceral endoderm, and then with parathyroid hormone-related peptide (PTHrP) to induce the transition to (mesenchymal) parietal endoderm. Here, we demonstrate that connexin43 mRNA and protein expression levels, protein phosphorylation and subcellular localization are dynamically regulated during F9 EC cell differentiation. Dye injection showed that this complex regulation of connexin43 is correlated with functional gap junctional communication. Similar patterns of connexin43 expression, localization and communication were found in visceral and parietal endoderm isolated ex vivo from mouse embryos at day 8.5 of gestation. However, in F9 cells this tightly regulated gap junctional communication does not appear to be required for the differentiation process as such. PMID- 10826496 TI - The zyxin-related protein TRIP6 interacts with PDZ motifs in the adaptor protein RIL and the protein tyrosine phosphatase PTP-BL. AB - The small adaptor protein RIL consists of two segments, the C-terminal LIM and the N-terminal PDZ domain, which mediate multiple protein-protein interactions. The RIL LIM domain can interact with PDZ domains in the protein tyrosine phosphatase PTP-BL and with the PDZ domain of RIL itself. Here, we describe and characterise the interaction of the RIL PDZ domain with the zyxin-related protein TRIP6, a protein containing three C-terminal LIM domains. The second LIM domain in TRIP6 is sufficient for a strong interaction with RIL. A weaker interaction with the third LIM domain in TRIP6, including the proper C-terminus, is also evident. TRIP6 also interacts with the second out of five PDZ motifs in PTP-BL. For this interaction to occur both the third LIM domain and the proper C-terminus are necessary. RNA expression analysis revealed overlapping patterns of expression for TRIP6, RIL and PTP-BL, most notably in tissues of epithelial origin. Furthermore, in transfected epithelial cells TRIP6 can be co-precipitated with RIL and PTP-BL PDZ polypeptides, and a co-localisation of TRIP6 and RIL with Factin structures is evident. Taken together, PTP-BL, RIL and TRIP6 may function as components of multi-protein complexes at actin-based sub-cellular structures. PMID- 10826497 TI - Spontaneous and induced apoptosis in embryogenic cell cultures of carrot (Daucus carota L.) in different physiological states. AB - Programmed cell death (apoptosis) in plants - and plant cells in culture - has received much less attention than its animal counterpart. In the present work, using agents producing biotic or abiotic stress on cultivated cells from carrot - and, in a few experiments, Arabidopsis -, we show that DNA fragmentation, random or oligonucleosomal, can be induced by different treatments. Moreover, we demonstrate that the same cultures may or may not respond to the inducing signal according to their physiological state. In particular, stationary cells are more responsive to the inducing signal than actively proliferating ones, and cells growing in an unorganized way are more responsive than cells carrying out the embryogenic programme. Senescent cells in culture also appear to die by apoptosis, but healthy cells can also be induced to die apoptotically if exposed to the medium conditioned by senescent cells of the same or different species. PMID- 10826498 TI - Regulation by carnitine of myocardial fatty acid and carbohydrate metabolism under normal and pathological conditions. AB - This review focuses on the regulation of myocardial fatty acids and glucose metabolism in physiological and pathological conditions, and the role of L carnitine and of its derivative, propionyl-L-carnitine. Fatty acids are the major oxidation fuel for the heart, while glucose and lactate provide the remaining need. Fatty acids in cytoplasm are transformed to long-chain acyl-CoA and transferred into the mitochondrial matrix by the action of three carnitine dependent enzymes to produce acetyl-CoA through the beta-oxidation pathway. Another source of mitochondrial acetyl-CoA is from the oxidation of carbohydrates. The pyruvate dehydrogenase (PDH) complex, the key irreversible rate limiting step in carbohydrate oxidation, is modulated by the intra mitochondrial ratio acetyl-CoA/CoA. An increased ratio results in the inhibition of PDH activity. A decreased ratio can relieve the inhibition of PDH as shown by the transfer of acetyl groups from acetyl-CoA to carnitine, forming acetylcarnitine, a reaction catalyzed by carnitine acetyl-transferase. This activity of L-carnitine in the modulation of the intramitochondrial acetyl CoA/CoA ratio affects glucose oxidation. Myocardial substrate metabolism during ischemia is dependent upon the severity of ischemia. A very severe reduction of blood flow causes a decrease of substrate flux through PDH. When perfusion is only partially reduced there is an increase in the rate of glycolysis and a switch from lactate uptake to lactate production. Tissue levels of acyl-CoA and long-chain acylcarnitine increase with important functional consequences on cell membranes. During reperfusion fatty acid oxidation quickly recovers as the prevailing source of energy, while pyruvate oxidation is inhibited. A considerable body of experimental evidence suggests that L-carnitine exert a protective effect in in vitro and in vivo models of heart ischemia and hypertrophy. Clinical trials confirm these beneficial effects although controversial results are observed. The actions of L-carnitine and propionyl-L carnitine cannot be explained as exclusively dependent on the stimulation of fatty acid oxidation but rather on a marked increase in glucose oxidation, via a relief of PDH inhibition caused by the elevated acetyl-CoA/CoA ratio. Enhanced pyruvate flux through PDH is beneficial for the cardiac cells since less pyruvate is converted to lactate, a metabolic step resulting in the acidification of the intracellular compartment. In addition, L-carnitine decreases tissue levels of acyl moieties, a mechanism particularly important in the ischemic phase. PMID- 10826499 TI - Defective suppression of the aldosterone biosynthesis during stroke permissive diet in the stroke-prone phenotype of the spontaneously hypertensive rat. AB - Previous studies have shown that short-term high salt intake unmasks blunted plasma aldosterone suppression in stroke-prone spontaneously hypertensive rats (SHRsp). The aim of this study was to evaluate the response of aldosterone biosynthesis and production to a sustained exposure to the stroke-permissive Japanese-style diet (JD) in young stroke-prone and stroke-resistant SHRs. For this purpose, 6-week old male rats from both strains were divided into 2 dietary groups and received regular diet (SHR = 37, SHRsp = 32) or the JD and 1% saline to drink (SHR = 34, SHRsp = 30) for 4 weeks. All measurements were carried out at the end of the dietary periods. After JD, plasma aldosterone levels were significantly decreased in SHR (from 357.8 +/- 57 to 163.3 +/- 31.5 pg/ml, p < 0.05) but markedly increased in SHRsp (from 442 +/- 56.5 to 739 +/- 125.7 pg/ml, p < 0.05). Consistently, the adrenal aldosterone synthase expression was reduced by JD in SHR (p < 0.05), whereas it was even slightly raised by JD in SHRsp so that, at the end of JD, aldosterone synthase mRNA was 5-fold higher in SHRsp than in SHR. Urinary sodium excretion (mEq/24h) achieved lower levels in SHRsp, so that fractional excretion of sodium was 80.2 +/- 9% in SHR and 40.3 +/- 8% in SHRsp (p < 0.05) in balance studies performed at the end of JD. These different responses of mineralocorticoid biosynthesis and urinary sodium excretion to JD were not accounted for by different adaptations of the renin-angiotensin and atrial natriuretic peptide systems, of serum potassium levels, or of adrenal 11beta-hydroxylase expression in the two strains. Systolic blood pressure was comparable in both strains throughout the experiment. These results demonstrate enhanced aldosterone biosynthesis, associated with reduced urinary excretion of sodium in response to JD in SHRsp before the onset of stroke. This abnormality may play a role in the higher susceptibility to stroke of this model. PMID- 10826500 TI - Mechanisms involved in coronary artery dilatation during respiratory acidosis in the isolated perfused rat heart. AB - A rat Langendorff heart preparation, perfused at constant pressure, was used to evaluate the role of K(ATP) channels in respiratory acidosis-induced coronary hyperemia. Prior administration of glibenclamide, an inhibitor of K(ATP) channels, reduced basal flow rates and eliminated the hyperemia associated with hypercapnia. These results implicate K(ATP) channels as a functional link in the respiratory acidosis-induced increase in coronary flow. PMID- 10826501 TI - Endothelin-1 induces interleukin-6 release via activation of the transcription factor NF-kappaB in human vascular smooth muscle cells. AB - The potent vasoconstrictor peptide endothelin-1 (ET-1) has been implicated in the pathophysiology of atherosclerosis and its complications. Since inflammation of the vessel wall is a hallmark of atherosclerosis, the purpose of the present study was to investigate the influence of ET-1 on cytokine production in human vascular smooth muscle cells (SMC) as a marker of inflammatory cell activation. ET-1 (100 pM - 1 microM) stimulated interleukin-6 (IL-6) secretion from human vascular SMC in a concentration-dependent manner. The ET-A-receptor antagonist BQ 123 (10 microM), but not the ET-B-receptor antagonist BQ-788, inhibited IL-6 release. ET-1 also transiently increased IL-6 mRNA compatible with regulation of IL-6 release at the pretranslational level. Electrophoretic mobility shift assays demonstrated time- and concentration-dependent activation of the proinflammatory transcription factor nuclear factor-kappaB (NF-kappaB) in ET-1-stimulated human vascular SMC. A decoy oligodeoxynucleotide bearing the NF-kappaB binding site inhibited ET-1-stimulated IL-6 release to a great extent suggesting that this transcription factor plays a key role for cytokine production elicited by ET-1. Moreover, the antioxidant pyrrolidine dithiocarbamate (10 microM) inhibited ET-1 induced IL-6 release indicating involvement of reactive oxygen species in ET-1 signaling. ET-1-stimulated IL-6 secretion was also suppressed by diphenylene iodonium (40 microM), an inhibitor of flavon-containing enzymes such as NADH/NADPH oxidase. The results demonstrate the ability of ET-1 to induce an inflammatory response in human vascular SMC. These observations may contribute to a better understanding of the role of ET-1 in inflammatory activation of the vessel wall during atherogenesis. PMID- 10826502 TI - Cardiac alpha-actin in smooth muscle cells: detection in umbilical cord vessels and in atherosclerotic lesions. AB - Phenotypic modulation of smooth muscle cells (SMC) is a key event during the development of atherosclerotic and restenotic lesions. During this process, the composition of the cytoskeleton is substantially altered, with changes predominantly in actin expression reflecting a shift from smooth muscle alpha actin to the non-muscle beta-isoform. We now demonstrate that yet another actin isoform, cardiac alpha-actin, is synthesized, de novo, in SMC of various atherosclerotic lesions. Using a highly specific monoclonal antibody against cardiac alpha-actin, we analyzed and compared the accumulation of this actin isoform in diverse SMC by immunofluorescence microscopy and immunoblotting. As expected, cardiac alpha-actin was present in human myocardium but not in healthy SMC of adult aorta, coronary arteries, trabeculae of the spleen, colon, stomach or skeletal muscle. Interestingly, the presence of cardiac alpha-actin was detected in umbilical cord vessels, human myometrium, in atherosclerotic coronary lesions and atherosclerotic lesions from peripheral vascular disease. The distribution of cardiac alpha-actin often paralleled that of cytokeratins 8 and 18, intermediate filament proteins typically found in dedifferentiated SMC. Taken together, the data presented here illustrate the expression of cardiac alpha actin to be limited to either fetal vessels or those vessels or tissue having suffered damage or atrophy, outside its 'native' environment in the heart. The demonstration of cardiac alpha-actin in SMC of umbilical cord vessels and in atherosclerotic lesions but not in apparently healthy vessels supports the notion that SMC in atherosclerotic lesions exhibit a dedifferentiated phenotype. PMID- 10826503 TI - Rp-cAMPS has no effect on adenosine A1 receptors in guinea-pig cardiomyocytes. AB - Rp-cAMPS, a protein kinase A inhibitor, is used in the investigation of the cAMP dependent systems. A report by Musgrave et al. has suggested that Rp-cAMPS may also act on adenosine receptors. To determine whether this occurs in guinea-pig ventricular myocytes, Rp-cAMPS was applied in the presence and absence of DCPCX, an adenosine A receptor antagonist. The isoprenaline-induced response was significantly decreased by Rp-cAMPS and the effect was not altered by the presence of DCPCX. Therefore Rp-cAMPS has no effect on cell contraction via adenosine A1 receptors and can reliably be used to investigate cyclic AMP dependent systems in isolated cardiac myocytes. PMID- 10826504 TI - Preconditioning-induced attenuation of purine metabolite accumulation during ischemia: memory and multiple cycles. AB - Brief myocardial ischemia (ISC) is known to attenuate purine metabolite accumulation in the interstitial fluid (ISF) during subsequent ISC. We determined how this attenuated purine accumulation was altered by 1) extended reperfusion (REP) and 2) multiple cycles of brief ISC. Microdialysis probes were used to assess ISF levels of the purine metabolites adenosine, inosine, and hypoxanthine in anesthetized rabbits. In one series of experiments, two 10 min periods of regional ISC were separated by 10 (n = 6), 60 (n = 6), or 180 (n = 6) min of REP. In the 10, 60, and 180 min REP groups the increase in ISF purine metabolites during the second ISC was 47%, 55%, and 53% of that seen during the first ISC, respectively. In a second series of experiments, hearts were exposed to 120 min of ISC with (n = 6) or without (n = 6) five preceding cycles of transient ISC (10 min ISC; 10 min of REP). The increase in ISF purine metabolites during the multiple cycles of ISC was progressively attenuated, and there was a delay but eventual increase in ISF purine metabolites during the 120 min ISC. These data demonstrate that attenuated purine metabolite accumulation 1) is progressively greater with multiple cycles of brief ISC, 2) has a memory time of at least 180 min, and 3) is not due to high energy phosphate depletion. PMID- 10826505 TI - Effects of preconditioning on myocardial interstitial levels of ATP and its catabolites during regional ischemia and reperfusion in the rat. AB - The interstitial accumulation of adenine nucleotide breakdown products (ANBP) in the myocardium during ischemia has been shown to provide a useful index of the ischemic injury, whereas reperfusion ANBP washout rate has been regarded as an index of reperfusion damage. The purpose of this study was, using cardiac microdialysis, to examine in the rat model of regional ischemia/reperfusion the relationship between the duration of ischemia and these indices and to assess the profile of interstitial ATP concentrations and the beneficial effects of ischemic preconditioning (IP). The rats underwent 10, 20, 30 or 40 min of coronary artery occlusion and 50 min of reperfusion. Regional ischemia, with its duration, provoked a progressive increase in dialysate ANBP in the ischemic zone. The rate of purine washout during reperfusion exponentially declined with an increase in duration of the ischemic period. IP, induced by three 5-min episodes of ischemia, each separated by 5 min of reperfusion, significantly reduced the accumulation of ANBP during the 30-min period of sustained ischemia and resulted in a marked acceleration of reperfusion ANBP washout, indicating the improvement of postischemic microcirculation. These effects were suggested to be, at least in part, responsible for the infarct size limitation observed. Using the relationship between the duration of ischemia and ANBP washout rate, it could be demonstrated that IP produced similar facilitation of purine washout as shortening of the ischemic period in nonpreconditioned rats from 30 to approximately 7 min. Regional 20-min ischemia induced an early peak increase in interstitial fluid ATP which correlated with the maximal incidence of ventricular arrhythmias, whereas IP abolished both ATP release and arrhythmias during the sustained ischemia. These findings suggest that ATP may be an important mediator of ischemia-induced ventricular arrhythmias. PMID- 10826506 TI - Protection of ischemic rat heart by dantrolene, an antagonist of the sarcoplasmic reticulum calcium release channel. AB - Cytosolic Ca2+ overload plays a major role in the development of irreversible injury during myocardial ischemia. Such overload is due at least in part to the release of Ca2+ from the sarcoplasmic reticulum. Therefore, we investigated whether dantrolene, a blocker of the sarcoplasmic reticulum Ca2+ release channel, may protect from ischemic injury. In binding experiments, we determined the effect of dantrolene on [3H]-ryanodine binding in rat cardiac tissue. In perfusion experiments, isolated rat hearts were perfused for 20 min in the working mode, in the presence of 0-45 microM dantrolene. The hearts were then subjected to 30 min of global ischemia and 120 min of retrograde reperfusion. Tissue injury was evaluated on the basis of triphenyltetrazolium chloride (TTC) staining and LDH release. The binding experiments showed that dantrolene displaced 4 nM [3H]-ryanodine with IC50 of 34 microM. In the perfusion experiments, tissue necrosis (i.e., TTC-negative tissue) averaged 28.3 +/- 1.6% of the ventricular mass under control conditions. Dantrolene was protective at micromolar concentrations: tissue necrosis decreased to 21.4 +/- 1.0% and 8.4 +/- 1.4% with 1 microM and 45 microM dantrolene, respectively (P < 0.05 and P < 0.01). Similar results were obtained with regard to LDH release. At low concentrations (up to 4 microM), dantrolene did not produce any significant hemodynamic effect, except for a slight increase in coronary flow, whereas at higher concentration a negative inotropic effect was apparent. In conclusion, dantrolene reduced ischemic injury even at concentrations that did not affect contractile performance. Modulation of sarcoplasmic reticulum Ca2+ release might represent a new cardioprotective strategy. PMID- 10826507 TI - Frequency-dependent changes in calcium cycling and contractile activation in SERCA2a transgenic mice. AB - OBJECTIVE: This study was undertaken to investigate the mechanism of altered contractility in hearts from transgenic mice overexpressing the sarcoplasmic reticulum (SR) Ca2+ ATPase (SERCA2a). In particular, we sought to determine whether the reported increase in contractility is frequency-dependent, as might be expected if attributable to changes in SR Ca2+ loading. METHODS: Intracellular [Ca2+] and contractile force were measured at room temperature (22 degrees C) simultaneously in fura-2-loaded isometrically-contracting trabeculae dissected from the hearts of FVB/N control (n = 6) or SERCA2a transgenic (n = 6) mice. RESULTS: SERCA transgenics exhibit a positive force-frequency relationship, but this was flat in age- and strain-matched controls. SERCA transgenics exhibit a sizable increase in calcium transient amplitude relative to controls, with a concomitant increase in force generation at higher frequencies of stimulation. Amplitudes of Ca2+ transients (transgenics: 1.56 +/- 0.09 micromol/L, controls: 1.21 +/- 0.14) and twitches (transgenics: 21.71 +/- 0.91 mN/mm2, controls: 13.74 +/- 1.67) were significantly different at 2.0 Hz stimulation (P < 0.05). CONCLUSION: An increase in SERCA expression increases the ability of the sarcoplasmic reticulum to store calcium, such that more calcium is available to be released during each heartbeat at higher stimulation rates. PMID- 10826508 TI - Cesium chloride induced ventricular arrhythmias in dogs: three-dimensional activation patterns and their relation to the cesium dose applied. AB - INTRODUCTION: Cesium chloride has widely been used in experimental models to produce various ventricular arrhythmias. The study was designed to evaluate whether type and mechanism of these arrhythmias are dose-dependent. METHODS: In 7 dogs with acute AV-block, 60 pins containing 4 bipolar electrodes each were inserted into both ventricles to provide 240 endo-, epi- and midmyocardial recording sites. A computerized mapping system was used to determine three dimensional activation patterns of ventricular arrhythmias induced by three injections of 1 mmol/kg cesium chloride at 20 minute intervals. RESULTS: Out of all arrhythmias induced, 25 ventricular extrasystoles, 31 monomorphic and 47 polymorphic ventricular tachycardias were mapped. Nonsustained ventricular tachycardias were readily inducible by a single bolus of cesium chloride, whereas sustained episodes required repetitive injections (1.45 +/- 0.61 vs. 2.61 +/- 0.57 doses, p < 0.05). Polymorphic tachycardias were observed more commonly than monomorphic tachycardias (87 vs. 31). Initiation and maintenance of cesium induced arrhythmias were exclusively based on focal mechanisms originating from the subendocardium, irrespective of morphology and dosage. All monomorphic arrhythmias were caused by repetitive firing of single immobile foci located in either the right or the left ventricle. Bi- and multifocal mechanisms, however, were found to underlie the polymorphic episodes. CONCLUSIONS: Although there is a dose-dependence as to the sustenance of mono- or polymorphic tachycardias, this does not reflect on the three-dimensional activation pattern of cesium induced arrhythmias, which are due to mono- or multifocal activation originating from the subendocardium. PMID- 10826509 TI - Analysis of mitochondrial DNA deletions in four chambers of failing human heart: hemodynamic stress, age, and disease are important factors. AB - Mitochondrial DNA (mtDNA) mutations are not only responsible for organ dysfunction due to inefficient energy production but also indicators of metabolic and functional stresses in the organ. To analyze the significance of deletion mutation in human myocardium, we screened the presence of two common deletions (7.4 kb from 8637-16084 nt, 5.0 kb from 8470-13477 nt) in four chambers using long-PCR, and using serial-dilution PCR, measured the amount of deleted mtDNA in normal heart (NL) of brain-dead victims of road accidents (n = 9, age = 10-59) and failing hearts (CHF) of patients who underwent heart transplantation (n = 24, age = 17-63). Frequency of both deletions was higher in ventricles (Vt) than in atria (At) (Vt:At = 25/33:12/33 for 7.4 kb, 19/33:6/33 for 5 kb) (p < 0.05), whereas it was the same in the right and left chambers. In ventricles, both deletions were more frequent among older persons (> 35 yrs) than in younger persons (< or = 35 yrs) (older:younger = 16/20:9/13 for 7.4 kb, 15/20:4/13 for 5 kb) (p < 0.05). In ventricles of failing heart, the 5-kb deletion was more frequent than in those of normal heart (CHF:NL = 17/24:2/9) (p < 0.05), whereas the 7.4-kb deletion was frequent both in failing and normal hearts (CHF:NL = 19/24:6/9). The association of mutation with aging or disease process observed in ventricles was not found in the atria. Although the amount of mutant mtDNA in the left ventricle tended to increase according to a disease process, it was small, at most 1.56% or 0.012% of total mtDNA for a 7.4- or 5-kb deletion, respectively. No deletion was found, however, in lymphocytes from any patient who underwent transplantation. In conclusion, deletion mutation of mtDNA is frequently, but in a small amount, found in the ventricle of older failing heart than in the atrium of younger normal heart. This suggests that hemodynamic stress, age, and disease are factors to induce mtDNA mutation that represents the indicator of stresses on the heart and might turn into a contributor of progressive heart failure under extreme conditions. PMID- 10826510 TI - Thyrocyte-interleukin-1 interactions. AB - Autoimmune thyroid disease is the most common organ-specific autoimmune disease and is a very common cause of thyroid dysfunction such as autoimmune hypothyroidism, Graves' disease and postpartum thyroiditis. The thyroid gland from patients with autoimmune thyroid disease is morphologically characterized by massive infiltration of lymphoid cells. The interleukin-1 (IL-1) family of molecules is together with other cytokines an integral component of the complex intercellular communication required to mount and control an immune response. IL 1alpha/beta in moderate and high concentrations reversibly inhibit thyroid cell function, while IL-1beta in low concentrations stimulates thyroid cell function. The biphasic, non-cytotoxic and reversible influence of IL-1 supports a role of IL-1 in the physiological regulation of thyroid cell function. IL-1 stimulates the guanylate mediated pathways and inhibits the adenylate cyclase mediated pathways. All IL-1 effects are counteracted by IL-1 receptor antagonist indicating that the effects are exerted through activation of specific IL-1 receptors on thyrocytes. Furthermore, IL-1 induces or enhances expression of a number of immunologically active molecules such as adhesion molecules, cytokines, and complement regulatory proteins in thyroid epithelial cells. IL-1 may thus play a role during physiological as well as pathophysiological conditions contributing to for example the euthyroid sick syndrome and development of thyroid autoimmunity. This review summarizes current literature on the phenomenological in vitro influence of IL-1 on the thyroid cell. PMID- 10826511 TI - Lack of germline mutations in the preproglucagon gene region coding for glucagon like peptide 1 in Type 2 diabetic (NIDDM) patients. AB - Glucagon-like peptide 1 (GLP-1) has antidiabetic effects and many facets of Type 2 diabetes could theoretically be the consequence of a reduction in or lack of GLP-1 function. Exogenous GLP-1 is exquisitely effective in Type 2 diabetic patients, making receptor defects unlikely. GLP-1 responses after meals as detected by radioimmunoassay are not overtly reduced in Type 2 diabetic patients. Therefore, a sequence analysis of exon 2 of the preproglucagon gene coding for the GLP-1 protein was initiated in order to exclude potential germline mutations. 24 Type 2 diabetic patients and in 14 control subjects with normal oral glucose tolerance (WHO criteria) were studied. In all specimens of peripheral blood leukocyte DNA examined, no germline mutations of the GLP-1 sequence were identified, thus excluding mutations in the GLP-1 sequence as a major contributor to the pathophysiological appearance of the Type 2 diabetic phenotype. Rare mutations, however, cannot be excluded due to the small number of Type 2 diabetic patients examined. PMID- 10826512 TI - Hemoglobin A1c and body mass index in children and adolescents with IDDM. An observational study from 1976-1995. AB - In adult patients with type 1 diabetes good metabolic control was associated with an undesired weight gain. In the present report the possible association of HbA1c and body mass index (BMI) in children and adolescents with type 1 diabetes (IDDM) was investigated in a long-term retrospective study from 1976 to 1995. Further, the relationship between BMI on one hand and age, gender, duration of IDDM, the number of units of insulin used and the number of injections per day on the other hand were considered. Statistical analysis was performed using repeated measurements analyses of variance. The 208 girls and 201 boys were 5-17 years old and had diabetes for beyond one year. For analysis 2512 data sets, in part measurements on the same patient in the course of the disease, were available. In various statistical models, the results show that age, gender, the daily amount of insulin, and the HbA1c level (p<0.001-0.005) were associated with the BMI. Extremely high HbA1c levels coincided with a remarkably low BMI. Hence, in children and adolescents with IDDM it may be difficult to achieve a constantly good metabolic control accompanied by a normal body weight. PMID- 10826513 TI - Maternally inherited diabetes and deafness (MIDD): unusual occult exocrine pancreatic manifestation in an affected German family. AB - The mitochondrial (mt) 3243 DNA mutation is an underlying cause of maternally inherited diabetes and deafness (MIDD) syndrome and the syndrome of mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes (MELAS). We report an affected German MIDD pedigree with maternal lineage over three generations. The index patient, her mother, her maternal aunt and her maternal grandmother all suffered from diabetes and premature hearing loss and were positive on testing for the mt 3243 DNA mutation. The 27-year-old index patient had a history of grand mal seizures. As sequela of abdominal ultrasound and confirmed by magnetic resonance cholangio-pancreaticography, she was diagnosed with chronic pancreatitis with pancreatic calcifications and pancreatic duct dilation, although she was completely asymptomatic and with no signs of steatorrhoea. She did not have gallstones and the common bile duct was normal. A possible etiopathogenic pathway for pancreatitis could be a suppressive effect of the mt 3243 mutation on the oxidative phosphorylation in affected mitochondria. Although pancreatitis and pancreatic dysfunction in association with the mt 3243 mutation, especially in patients with comorbidity of MELAS and diabetes, has previously been described as a rare manifestation, this case is specific because of the discrepancy of advanced morphological pancreatic alterations and complete lack of pancreatogenic symptoms. PMID- 10826514 TI - A study on the genetics of obesity: influence of polymorphisms of the beta-3 adrenergic receptor and insulin receptor substrate 1 in relation to weight loss, waist to hip ratio and frequencies of common cardiovascular risk factors. AB - Beta-3-adrenergic receptor (beta-3-AR) and insulin receptor substrate 1 (IRS-1) have been implicated in the pathogenesis of obesity and in obesity related increase in insulin resistance which is associated with, among other diseases, dyslipidemia and type 2 diabetes mellitus. We studied 210 white female Caucasian obese subjects, who underwent a formal weight loss program (Optifast). We examined the association between mutations of the IRS-1 gene at codon 972, mutations of the beta-3-AR gene at codon 64, and the combination of both mutations with the degree of weight loss, waist to hip ratio and the prevalence of hypertension, dyslipidemia and type 2 diabetes mellitus. Twenty-four women (11.4%) were polymorph only for the beta-3-AR mutation, 23 women (10.9%) only for the IRS-1 mutation, and 6 subjects (2.9%) were polymorph for both alleles. No patient displayed a homozygous polymorphism. Similar frequencies of these polymorphisms were observed when the 100 non-obese control women were tested (14.0, 15.0, 3.0, respectively). After 13 weeks of weight loss the group with multiple polymorph alleles had lost less of their weight than the obese controls without mutation (Delta BMI 5.32+/-0.18 versus 6.12+/-0.2 kg/m2, p<0.05). In this group, the frequency of type 2 diabetes (66.7%) was significantly higher than in the obese control group without mutations (16.7%, p=0.008). Our findings suggest there is a synergy between the polymorphisms of Trp64Arg beta-3-AR and Gly972Arg IRS-1 in Caucasian German obese women leading to a decreased weight loss. This seems to be accompanied with an increased frequency of type 2 diabetes. PMID- 10826515 TI - Prevalence and atherosclerosis risk in different types of non-diabetic hyperglycemia. Is mild hyperglycemia an underestimated evil? AB - So far little is known about the importance of different types of non-diabetic hyperglycemia for the development of macrovascular disease. The aim of this work was to examine the intima-media thickness (IMT) of the common carotid artery (CCA), a well-accepted marker of atherosclerosis, as well as various risk factors for atherosclerosis in non-diabetic subjects with isolated fasting (IFH; n=67), isolated postchallenge (IPH; n=82) and combined hyperglycemia (CH; n=88) in comparison to normoglycemic (NG; n=265) controls. Subjects were participants of the RIAD study (Risk Factors in IGT for Atherosclerosis and Diabetes). IMT in the IPH (IMTmean: 0.89+/-0.02 mm; IMTmax: 1.01+/-0.02 mm; mean+/-SEM) and CH group (IMTmean: 0.91+/-0.02 mm; IMTmax: 1.03+/-0.02 mm) was significantly increased vs. the NG (IMTmean: 0.82+/-0.01 mm; IMTmax: 0.94+/-0.01 mm) and IFH group (IMTmean: 0.81+/-0.02 mm; IMTmax: 0.90+/-0.03 mm). IMT of the IFH group was similar to the normoglycemic controls. Subjects in the first and second tertile for postchallenge plasma glucose have similar carotid IMT irrespective of the level of fasting plasma glucose. The individuals of the third tertile for 2 h plasma glucose, whether in the first, second or third tertile of fasting plasma glucose, showed the same carotid IMT, which was significantly higher than all other groups, except for the one with lowest tertile for fasting and postchallenge plasma glucose. Except for total cholesterol and von Willebrand factor the levels of all other risk parameters were significantly higher in the hyperglycemic groups in comparison to the normoglycemic controls. Among the hyperglycemic subjects the CH group was at the highest risk for atherosclerosis with significantly increased levels of plasma triglycerides, fibrinogen, PAI-1, albuminuria, HDL-triglycerides, free fatty acids, insulin and proinsulin, and significantly reduced HDL-cholesterol in comparison to the normoglycemic controls. In summary, postchallenge hyperglycemia within the non-diabetic range is associated with atherosclerosis, as measured by the increased intima-media thickness of the common carotid artery. Furthermore, cardiovascular risk factors are significantly raised in all types of non-diabetic hyperglycemia. PMID- 10826516 TI - Pharmacokinetic and pharmacodynamic properties of long-acting insulin analogue NN304 in comparison to NPH insulin in humans. AB - NN304 is a long-acting insulin analogue that is acylated with a 14-C-fatty acid chain. Protraction of action of this novel insulin analogue is due not to slow absorption after subcutaneous administration but to reversible binding to albumin. We investigated the pharmacokinetic and pharmacodynamic properties of insulin analogue NN304 (0.3 and 0.6 U/kg) in comparison to NPH insulin (0.3 and 0.6 IU/kg) in 10 healthy volunteers performing a randomised, double-blind, cross over, placebo-controlled glucose clamp study. During the observation period of 24 hours the areas under the insulin curve for NPH[0.3 IU/kg] vs. NPH[0.6 IU/kg] were 60 vs. 102 nmol min l(-1) (p<0.01) and for insulin analogue NN304[0.3 U/kg] vs. NN304[0.6 U/kg] 490 vs. 932 nmol min l(-1) (p <0.001), suggesting a clear dose-response relationship for both NPH insulin and NN304. The amount of disposed glucose (area under the curve of glucose infusion) differed with statistical significance between the five treatments and was highest with NPH[0.6 IU/kg] (2671 mg/kg) and lowest with placebo (265 mg/kg). However, area under the curve of glucose infusion after treatment with NN304 was only 36% (dose of 0.3 U/kg) and 24% (dose of 0.6 U/kg) of that observed with corresponding doses of NPH insulin. Moreover, increasing dosages of NN304 failed to demonstrate a significant dose-response with regard to the area under the curve of glucose infusion. This study demonstrates that the principle of protracted insulin action of NN304 by reversible binding to albumin is effective in humans albeit at a much lower rate of glucose utilisation when compared to NPH insulin. Thus, in contrast to animal studies NN304 and NPH insulin can not be considered equipotent in humans. PMID- 10826517 TI - Increased levels of platelet-derived growth factor in vitreous fluid of patients with proliferative diabetic retinopathy. AB - It is generally accepted that growth factors play an important role in the pathogenesis of proliferative diabetic retinopathy. Since platelet-derived growth factor AB (PDGF AB) is known to be involved in many angiogenetic and proliferative processes, it was the aim of our study to elucidate the role of PDGF AB in the angiogenetic process in proliferative diabetic retinopathy. We measured PDGF AB concentrations in the vitreous of 23 patients with proliferative diabetic retinopathy, 4 of them with additional rubeosis iridis as an indicator of very high vasoproliferative activity. Control measurements were done in 19 patients without diabetic or ischemic eye diseases and also in 4 non-diabetic patients with ischemic proliferative retinopathy with rubeosis iridis. To exclude PDGF remnants in the vitreous due to vitreous bleeding we additionally measured platelet factor 4 concentrations as a stable marker of activated thrombocytes in the vitreous. RESULTS: Significantly elevated concentrations of PDGF AB were found in the vitreous of patients with proliferative diabetic retinopathy, with higher levels in individuals with additional rubeosis iridis compared to controls. However, concentrations of PDGF AB were also elevated in ischemic non diabetic retinopathy, supporting the concept that ischemia might be a strong stimulator of growth factor production in the retina. Platelet factor 4 was not detectable in any of the vitreous samples included in the study. In summary, our results indicate that the growth factor PDGF plays an important role in the pathogenesis of proliferative diabetic retinopathy, probably in synergistic action with other growth factors like IGF I, IGF II, VEGF and TNF alpha. PMID- 10826518 TI - Detection of functionally different types of pathological autoantibodies against thyrotropin receptor in Graves' patients sera by luminescent immunoprecipitation analysis. AB - We describe a new method for the detection of different types of pathological autoantibodies against TSH receptor (TSHR) in Graves' patients sera by luminescent immunoprecipitation analysis. For this purpose three different chimeras composed of human TSHR and rat luteotropin/choriogonadotropin receptor (LH-CGR) were constructed, as was described previously (Tahara K, Ishikawa N, Yamamoto K, Hirai A, Ito K, Tamura Y, Yoshida S, Saito Y, Kohn LD. 1997 Thyroid 7:867-877). They were used in the immunoprecipitation reactions: (i) the wild type TSHR (for the detection of total TSHR autoantibodies), (ii) TSHR/LH-CGR chimera wherein TSHR amino acid residues 8-165 (epitopes for thyroid stimulating antibodies) are replaced by comparable LH-CGR residues, (iii) TSHR/LH-CGR chimera wherein TSHR amino acids 261-370 (epitopes for thyroid blocking antibodies) are replaced by comparable LH-CGR residues, and (iv) TSHR/LH-CGR chimera wherein TSHR amino acids 8-165 and 261-370 are replaced by comparable LH-CGR residues (for the detection of neutral TSHR autoantibodies). DNA encoding the N-terminal 725 (of 764) amino acids of wild type TSHR (or TSHR/LH-CGR chimera) was fused to the cDNA for the 550-amino acid firefly luciferase. The hybrid proteins were produced in HeLa cells using recombinant vaccinia viruses. All fusion proteins retained the enzymatic activity of firefly luciferase and TSHR-LUC interacted with TSH with the same affinity as wild type receptor. The luciferase tagged TSHR and TSHR/LH CGR chimeras were used for the detection of different types of TSHR autoantibodies (i.e. total, neutral, thyroid stimulating and thyroid blocking) in 63 Graves' disease and 62 normal sera by immunoprecipitation analysis. The data demonstrated positive correlation between results of immunoprecipitation assay and results obtained using cAMP bioassay or assay for TSH binding inhibitory immunoglobulins in test sera. PMID- 10826519 TI - Characterization of the neuroprotective effects of estrogens on hydrogen peroxide induced cell death in hippocampal HT22 cells: time and dose-dependency. AB - Time and dose-dependency of the effects of estrogens (17-beta estradiol, estrone) and non-estrogenic steroids (progesterone, dexamethasone and methylprednisolone) on the toxicity of hydrogen peroxide were examined in mouse hippocampal HT22 cells. Hydrogen peroxide, an important intermediate of various disease-relevant oxidative stressors, induced cell death in HT22 cells in extracellular concentrations between 0.5 and 1.5 mM in a dose-dependent manner (EC50=0.95 mM). Regarding the underlying mechanisms of toxicity, incubation with hydrogen peroxide did not induce lipid peroxidation in living HT22 cells under these conditions. After preincubation with estrogens and non-estrogenic steroids for 22 hours, estrogen compounds protected the cells against hydrogen peroxide toxicity. Estrogens showed a maximal protective effect at 60-70% of hydrogen peroxide toxicity which diminished at higher and lower concentrations of the toxic challenge. Dose-dependency studies of estrogens revealed that concentrations of 1 microM already exerted a significant cytoprotective effect. Co- and postincubation with 17-beta estradiol and estrone also resulted in significant cell protection even if the estrogens were added 30 min after the initiation of the challenge with hydrogen peroxide. In contrast, preincubation with other steroids like progesterone, a physiological gonadal steroid, dexamethasone, a synthetic glucocorticoid and methylprednisolone, a glucocorticoid with radical scavenging properties, did not protect the cells against hydrogen peroxide toxicity but resulted in a dose-related decrease of HT22 cell survival in the course of the toxic challenge. PMID- 10826520 TI - A RET double mutation in the germline of a kindred with FMTC. AB - Activating germline mutations of the RET proto-oncogene are found in more than 90% of families with multiple endocrine neoplasia type 2a (MEN 2a) and familial medullary thyroid carcinoma (FMTC). The majority of patients with these hereditary tumors carry germline mutations that result in the substitution of one of five cysteine residues in exon 10 and 11. Different mutations in exons 13, 14 and 15 affecting non-cysteine residues have also been described but are considered to be rare. We now for the first time report a double mutation of the RET proto-oncogene occurring in the germline of a kindred with FMTC. Both mutations occur within the tyrosine kinase domain in exon 14 and lead to the substitution of valine 804 by methionine and arginine 844 by leucine. Since the double mutated allele cosegregated with the disease and was not identified in 200 unrelated normal probands, we conclude that they represent mutations that predispose the individual to the development of FMTC with a mild phenotype. PMID- 10826521 TI - Radioiodine therapy in Graves' hyperthyroidism: determination of individual optimum target dose. AB - The aim of this study was to investigate the results of different volume dependent target doses on clinical outcome 6 months after radioiodine therapy (RITh) and its correlation with post therapeutic thyroid volumes (Vpost) in patients with Graves' disease. This analysis was designed to determine factors improving the results of radioiodine therapy without increasing target doses generally, as has been recommended recently. We studied consecutive data from 102 patients with Graves' disease, who had initial radioiodine therapy between 1991 and 1995. The 131I activities were calculated according to the formula of Marinelli. In addition to the normal calculation individual target doses were adjusted to the thyroid volumes of each patient before therapy. For statistical evaluation, the patients were divided into three subgroups of comparable sample sizes: Group I included those with a thyroid volume <15 ml before therapy. Group II included those ranging from a 15-25 ml volume before therapy and group III included those with thyroid volumes >25 ml. Laboratory thyroid parameters and thyroid volumes were measured in those groups before and 6 months after therapy. RESULTS: Analysis of all patients revealed a significantly higher rate of hypothyroidism (54%) and fewer cases of hyperthyroidism (15%) six months after therapy in cases with Vpost smaller than 8 ml. The median Vpost needed to achieve an optimum therapeutic success rate (rate of eu- or hypothyroidism) was smaller than 5 ml in group I and smaller than 10 ml in group II. Therapeutic success was associated with different target doses in each group, 150,220 and 260 Gy for groups I, II, and III respectively. CONCLUSIONS: Post therapeutic thyroid volumes correlated significantly with clinical outcome six months after therapy. An adjustment of the target doses based on thyroid volumes before therapy will lead to an appropriate reduction of thyroid volumes. Thus, in the individual case clinical outcome could be improved without applying higher target doses in all patients. This would ensure a better utilization of limited resources in medical care e.g. through a shorter hospital stay. PMID- 10826522 TI - Decreased thyroid peroxidase expression in cultured thyrocytes after external gamma irradiation. AB - Impairment of thyroid function has been described in up to 50% of the patients after external irradiation of the neck region as well as after mantle irradiation. In order to assess radiation-induced alterations in cultured thyroid cells, the occurrence of apoptosis and necrosis as well as the expression of thyroid peroxidase (TPO) and of two members of the 70 kD heat shock family, HSP 73 and HSP-72, were analysed following gamma irradiation. Human thyroid epithelial cells (TEC) were purified from surgical tissue specimens, were cultured and irradiated with a single dose of 5 Gy or 50 Gy using Co60 as radioactive source. Analysis was performed 1, 3 and 5 day(s) after irradiation. Apoptosis and necrosis were assessed by DNA staining with propidium iodide and FACS analysis. TPO and HSP expression by SDS-PAGE and Western blotting. The cell viability of TEC was not affected by irradiation and there was no induction of HSP-72, a sensitive indicator of acute cellular stress. Interestingly, the expression of TPO, a key enzyme of thyroid hormone synthesis, decreased significantly in irradiated TEC, while HSP-73 expression remained unchanged. Decreased expression of TPO with a resulting suppression of thyroid hormone synthesis could contribute to an early development of thyroid dysfunction following irradiation. Thus, analysis of thyroid function, even early after external radiation therapy of the neck or after total body irradiation, seems to be indicated. PMID- 10826523 TI - Melatonin hypersecretion in male patients with adult-onset idiopathic hypogonadotropic hypogonadism. AB - Increased melatonin secretion observed in male patients with congenital isolated hypogonadotropic hypogonadism and its normalization during testosterone treatment had suggested that melatonin and the reproductive hormones are inter-related. Since these patients have a congenital form of hypogonadism, it is likely that hypermelatoninemia is the consequence of hypogonadism. To further study the relations between the pineal and the reproductive axis in humans, we evaluated melatonin secretion in two men (aged 35 and 50 yrs.) with acquired adult-onset hypogonadotropic hypogonadism. The diagnosis was based on the findings of normal testicular volume, azoospermia, low serum testosterone, normal LH and FSH levels, but apulsatile LH secretion, and intact anterior pituitary hormones secretion, normal findings on skull radiographic imaging, prior sexual maturation and paternity. Melatonin secretion was assessed as urinary 24 h 6-sulphatoxymelatonin excretion (aMT6s) prior to and during the administration of 250 mg testosterone enanthate per month for 4 months. Pretreatment melatonin production was markedly increased in both patients: 427-915 ng/kg/24 h vs. 204+/-81 [mean+/-SD] in 16 age matched male controls. During testosterone treatment, aMT6s levels were normalized in one patient (range: 81-287 ng/kg/24 h) and remained elevated in the other patient (range: 830-1280 ng/kg/24 h). These data indicate that male patients with acquired GnRH deficiency have increased melatonin secretion. Melatonin hypersecretion in these patients may reflect a functional association. PMID- 10826524 TI - Pneumocystis carinii pneumonia associated with a rapid reduction of cortisol level in a patient with ectopic ACTH syndrome treated by octreotide and ketoconazole. AB - A case is herein reported of pneumocystis carinii pneumonia in a 60-year-old female patient with ectopic production of ACTH at a position 2 cm superior to her right clavicle, revealed in an octreotide scan. Her extremely high plasma ACTH and cortisol levels (460 pg/ml and 80 microg/dl, respectively) were markedly decreased with the combined treatment of octreotide (300 microg/d) and ketoconazole (600 mg/d). As her serum cortisol concentration decreased, pneumocystis carinii pneumonia occurred on the third day of treatment. A secondary E. coli infection was superimposed and the patient died of disseminated intravascular coagulation and adult respiratory distress syndrome. This case suggests that primary prophylaxis for pneumocystis carinii infection should be initiated before cortisol lowering therapy, especially when the plasma cortisol concentration is excessively high, and that early adjunctive glucocorticoid therapy can reduce the acute mortality in patients with endogenous Cushing's syndrome and Pneumocystis carinii pneumonia. This case study would also like to point out that plasma ACTH and cortisol levels were decreased effectively by the combination of octreotide and ketoconazole in this instance of ectopic ACTH syndrome. PMID- 10826525 TI - Serine protease inhibitors: novel therapeutic targets for stroke? AB - Although the thrombolytic activity of tissue-type plasminogen activator (t-PA) may be beneficial in the acute treatment of stroke, recent studies have suggested that this serine protease could also play a critical role in determining the extent of neuronal death after injury to the central nervous system (CNS). This hypothesis is based on several experimental results: t-PA-deficient mice are resistant to excitotoxic neuronal death induced by the intrahippocampal injection of kainate; the infarct volume induced by occlusion of the middle cerebral artery is reduced in t-PA knockout mice; and the intravenous injection of t-PA can under certain circumstances potentiate the infarct volume in animals subjected to middle cerebral artery occlusion. In the CNS, the serine proteases have been identified to occur both in neurons and glial cells. Their enzymatic activity regulates the balance between the accumulation and the degradation of the extracellular matrix. They are involved in many physiologic functions, ranging from synaptic outgrowth during perinatal development to plasticity in adults. For instance, thrombin and t-PA are known to modulate neurite outgrowth and tissue remodeling in the early stages of development. In the adult brain, t-PA may contribute to the late phase of long-term potentiation and to the subsequent synaptic growth in the hippocampal mossy fiber pathway. This balance between the degradation and accumulation of the extracellular matrix may also be integral to various pathologic processes involved in acute brain injury. For example, compounds that modulate the activity of serine proteases exhibit neuroprotective activity. Based on the above, numerous studies have focused on the production and modulation of the endogenously produced serine protease inhibitors, termed serpins, such as type 1 plasminogen activator inhibitor, neuroserpin, and protease nexin-1. In the present review, we will discuss the need to distinguish between the potentially neurotoxic effects of t-PA and its beneficial effect on reperfusion. We will present data supporting the idea that the modulation of serine protease activity may represent a novel and efficient strategy for the treatment of acute cerebral injury in humans. PMID- 10826526 TI - Quantitative assessment of ischemic pathology in axons, oligodendrocytes, and neurons: attenuation of damage after transient ischemia. AB - Axons and oligodendrocytes are vulnerable to cerebral ischemia. The absence of quantitative methods for assessment of white matter pathology in ischemia has precluded in vivo evaluation of therapeutic interventions directed at axons and oligodendrocytes. The authors demonstrate here that the quantitative extent of white matter pathology was reduced by restoration of cerebral blood flow after 2 hours of middle cerebral artery occlusion. Focal ischemia was induced in anesthetized rats by intraluminal thread placement, either transiently (for 2 hours) or permanently. At 24 hours after induction of ischemia, axonal damage was determined by amyloid precursor protein (APP) immunohistochemistry, and the ischemic insult to oligodendrocytes was assessed by Tau-1 immunostaining in the same sections. In adjacent sections, ischemic damage to neuronal perikarya was defined histologically. The hemispheric extent of axonal damage was reduced by 70% in the transiently occluded animals from that in permanently occluded animals. The volumes of oligodendrocyte pathology and of neuronal perikaryal damage were reduced by 62% and 58%, respectively, in the transiently occluded animals. These results demonstrate that this methodologic approach for assessing ischemic damage in axons and oligodendrocytes can detect relative alterations in gray and white matter pathology with intervention strategies. PMID- 10826527 TI - NMDA receptor blockade fails to alter axonal injury in focal cerebral ischemia. AB - The ability of the NMDA receptor antagonist, MK-801, to protect myelinated axons after focal cerebral ischemia has been examined. Amyloid precursor protein (APP) immunocytochemistry was used to assess the anatomic extent of axonal injury, and conventional histopathology was used to assess the volume of ischemic damage to neuronal perikarya. The middle cerebral artery was permanently occluded in 16 cats. The cats were treated with either vehicle or MK-801 as a 0.5-mg/kg bolus at 15 minutes before middle cerebral artery occlusion, followed by an infusion of 0.14 mg/kg per hour. After 6 hours, the animals were killed and the brains processed for histology and immunocytochemistry. The volume of neuronal necrosis was determined from 16 preselected coronal levels of the brain. The circumscribed zones of APP accumulation in axons were mapped onto images at the same 16 coronal levels, and quantitative analysis was performed using a transparent counting grid, randomly placed over each image. The histologic appearance and anatomic location of axons with increased APP immunoreactivity was similar in animals treated with vehicle and MK-801. MK-801 failed to reduce the hemispheric APP score significantly. In vehicle-treated animals, there was a significant association between the volume of neuronal necrosis and the amount of APP immunoreactivity. MK-801 significantly reduced the slope of the association between the volume of neuronal necrosis and the amount of APP immunoreactivity compared with that observed in vehicle-treated animals. As a result, the ratio of hemispheric APP score and volume of neuronal necrosis was significantly increased with MK-801 treatment. The inability of NMDA receptor antagonists to protect axons may limit their functional efficacy in improving functional outcome after stroke. PMID- 10826528 TI - Correlation between N-acetylaspartate levels and histopathologic changes in cortical infarcts of mice after middle cerebral artery occlusion. AB - The aim of the present study was to evaluate the use of the endogenous neuronal compound N-acetylaspartate (NAA) as a marker of neuronal damage after focal cerebral ischemia in mice. After occlusion of the middle cerebral artery (MCAO) the ischemic cortex was sampled, guided by 2,3,5-triphenyltetrazolium chloride (TTC) staining, and the NAA concentration was measured by high-pressure liquid chromatography (HPLC). Conventional histology and immunohistological methods using antibodies against neuron-specific enolase (NSE), neurofilaments (NF), synaptophysin, glial fibrillary acidic protein (GFAP), and carbodiamide-linked NAA and N-acetylaspartylglutamate (NAAG). The level of NAA rapidly declined to 50% and 20% of control levels in infarcted tissue after 6 hours and 24 hours, respectively. No further decrease was observed during the observation period of 1 week. Within the first 6 hours the number of normal-appearing neurons in the infarcted cortical tissue decreased to 70% of control, of which the majority were eosinophilic. After 24 hours almost no normal-appearing neurons were seen. The number of eosinophilic neurons decreased steadily to virtually zero after 7 days. The number of immunopositive cells in the NSE, NF, and synaptophysin staining within the infarct was progressively reduced, and after 3 to 7 days the immunoreactions were confined to discrete granulomatous structures in the center of the infarct, which otherwise was infested with macrophages. This granulomatous material also stained positive for NAA. The number of cells with positive GFAP immunoreactions progressively increased in the circumference of the infarct. They also showed increased immunoreaction against NAA and NSE. The study shows that the level of NAA 7 days after ischemia does not decline to zero but remains at 10% to 20% of control values. The fact NAA is trapped in cell debris and NAA immunoreactivity is observed in the peri-infarct areas restricts its use as a marker of neuronal density. PMID- 10826529 TI - Early N-acetylaspartate depletion is a marker of neuronal dysfunction in rats and primates chronically treated with the mitochondrial toxin 3-nitropropionic acid. AB - N-acetylaspartate (NAA) quantification by 1H-magnetic resonance spectroscopy has been commonly used to assess in vivo neuronal loss in neurodegenerative disorders. Here. the authors used ex vivo and in vivo 1H-magnetic resonance spectroscopy in rat and primate models of progressive striatal degeneration induced by the mitochondrial toxin 3-nitropropionate (3NP) to determine whether early NAA depletions could also be associated with neuronal dysfunction. In rats that were treated for 3 days with 3NP and had motor symptoms, the authors found a significant decrease in NAA concentrations, specifically restricted to the striatum. No cell loss or dying cells were found at this stage in these animals. After 5 days of 3NP treatment, a further decrease in striatal NAA concentrations was observed in association with the occurrence of dying neurons in the dorsolateral striatum. In 3NP-treated primates, a similar striatal-selective and early decrease in NAA concentrations was observed after only a few weeks of neurotoxic treatment, without any sign of ongoing cell death. This early decrease in striatal NAA was partially reversed after 4 weeks of 3NP withdrawal. These results demonstrate that early NAA depletions reflect a reversible state of neuronal dysfunction preceding cell degeneration and suggest that in vivo quantification of NAA 1H-magnetic resonance spectroscopy may become a valuable tool for assessing early neuronal dysfunction and the effects of potential neuroprotective therapies in neurodegenerative disorders. PMID- 10826530 TI - The heat shock response inhibits NF-kappaB activation, nitric oxide synthase type 2 expression, and macrophage/microglial activation in brain. AB - The heat shock response (HSR) provides protection against stress-induced damage, and also prevents initiation of inflammatory gene expression via inhibition of NFkappaB activation. This article describes experiments demonstrating that the HSR prevents induction of nitric oxide synthase type 2 (NOS2) in rat brain. Twenty four hours after intrastriatal injection of lipopolysaccharide (LPS), IL 1beta, and IFN-gamma, NOS2 immunoreactive cells were detected in striatum, corpus callosum, and to a lesser extent in cortex. Induction of a HSR by whole body warming to 41 degrees C for 20 minutes, done 1 day before LPS plus cytokine injection, reduced the number of NOS2-positive staining cells to background levels. Staining for EDI antigen revealed that the HSR also suppressed microglial/brain macrophage activation in the same areas. Striatal injection of LPS and cytokines induced the rapid activation of NFkappaB, and this activation was prevented by prior HS, which also increased brain IkappaB-alpha expression. These results suggest that establishment of a HSR can reduce inflammatory gene expression in brain, mediated by inhibition of NFkappaB activation, and may therefore offer a novel approach to treatment and prevention of neurological disease and trauma. PMID- 10826531 TI - Modulation by nitric oxide of cerebral neutrophil accumulation after transient focal ischemia in rats. AB - A beneficial role of nitric oxide (NO) after cerebral ischemia has been previously attributed to its vascular effects. Recent data indicate a regulatory role for NO in initial leukocyte-endothelial interactions in the cerebral microcirculation under basal and ischemic conditions. In this study, the authors tested the hypothesis that endogenous NO production during and/or after transient focal cerebral ischemia can also be neuroprotective by limiting the process of neutrophil infiltration and its deleterious consequences. Male Sprague-Dawley rats were subjected to 2 hours occlusion of the left middle cerebral artery and the left common carotid artery. The effect of NG-nitro-L-arginine methyl ester (L NAME) (10 mg/kg, intraperitoneally), an NO synthase inhibitor, was examined at 48 hours after ischemia on both infarct size and myeloperoxidase activity, an index of neutrophil infiltration. L-NAME given 5 minutes after the onset of ischemia increased the cortical infarct volume by 34% and increased cortical myeloperoxidase activity by 60%, whereas administration of L-NAME at 1, 7, and 22 hours of reperfusion had no effect. Such exacerbations of infarction and myeloperoxidase activity produced when L-NAME was given 5 minutes after the onset of ischemia were not observed in rats rendered neutropenic by vinblastine. These results suggest that after transient focal ischemia, early NO production exerts a neuroprotective effect by modulating neutrophil infiltration. PMID- 10826532 TI - L-arginine partially restores the diminished CO2 reactivity after mild controlled cortical impact injury in the adult rat. AB - Using an open cranial window technique, the authors investigated the mechanisms associated with the suppressed CO2 reactivity after mild controlled cortical impact (CCI) injury in rats. The dilation of arterioles (n = 7) to hypercapnia before injury was 38 +/- 12%, which was significantly reduced both at 1 hour (23 +/- 15% dilation) and at 2 hours after injury (11 +/- 19% dilation). In the presence of L-arginine (10 mmol/L topical suffusion, 300 mg/kg intravenous infusion), the dilation of pial arterioles (n = 6) to hypercapnia was partially restored to 30 +/- 6% at 2 hours after injury. In the presence of the nitric oxide (NO) donor, S-nitroso-N-acetylpenicillamine (SNAP) (10(-8) mol/L topical suffusion), the dilation of pial arterioles (n = 5) to hypercapnia remained diminished (5 +/- 7%) at 2 hours after injury. The dilation of pial arterioles (n = 4) to hypercapnia also remained suppressed (5 +/- 6%) with topical suffusion of the free radical scavengers, polyethylene glycol-superoxide dismutase (60 units/mL) and polyethylene glycol-catalase (40 units/mL). The authors have shown that L-arginine at least partially restores the diminished response to hypercapnia after mild CCI injury. Furthermore, these data suggest that the beneficial effects of L-arginine are mediated by a combination of providing substrate for NO synthase and scavenging free radicals. PMID- 10826533 TI - Excitotoxic hippocampal injury is attenuated in thioredoxin transgenic mice. AB - Thioredoxin is a small, multifunctional protein with a redox-active disulfide/dithiol in the active site. Thioredoxin plays several important biologic roles both in intracellular and extracellular compartments with its redox-regulating and reactive oxygen intermediates scavenging activities. We assayed the seizure response and the excitotoxic hippocampal injury in thioredoxin transgenic and wild-type C57BL/6 mice. Seizure score after kainic acid treatment was significantly lower in thioredoxin transgenic mice. Seven days after kainic acid administration, the damage in the hippocampal CA1 and CA3 regions was significantly attenuated in thioredoxin transgenic mice. Thioredoxin and redox regulation play an important role in excitotoxic brain damage. PMID- 10826534 TI - Induced hypothermia in experimental pneumococcal meningitis. AB - Pneumococcal meningitis resulting from Streptococcus pneumoniae has a death rate of 28% in adults. In severe head injury and stroke, inflammatory changes and intracranial hypertension are improved by induced hypothermia, which also is neuroprotective. We hypothesized that moderate hypothermia ameliorates inflammatory changes in experimental pneumococcal meningitis. Wistar rats were cooled systemically, and meningitis was induced by pneumococcal cell wall components. The increase of regional cerebral blood flow in the meningitis animals was blocked by hypothermia at 6 hours. The reduction of intracranial pressure correlated with temperature. The influx of leukocytes into the cerebrospinal fluid and levels of tumor necrosis factor alpha in the cerebrospinal fluid were decreased. Cooling the animals 2 hours after meningitis induction to 30.5 degrees C was also protective. We conclude that hypothermia is a new adjuvant approach to reduce meningitis-induced changes, in particular intracranial pressure, in the early phase of the disease. PMID- 10826535 TI - Involvement of intracellular calcium stores during oxygen/glucose deprivation in striatal large aspiny interneurons. AB - Striatal large aspiny interneurons were recorded from a slice preparation using a combined electrophysiologic and microfluorometric approach. The role of intracellular Ca2+ stores was analyzed during combined oxygen/glucose deprivation (OGD). Before addressing the role of the stores during energy deprivation, the authors investigated their function under physiologic conditions. Trains of depolarizing current pulses caused bursts of action potentials coupled to transient increases in intracellular calcium concentration ([Ca2+]i). In the presence of cyclopiazonic acid (30 micromol/L), a selective inhibitor of the sarcoendoplasmic reticulum Ca2+ pumps, or when ryanodine receptors were directly blocked with ryanodine (20 [micromol/L), the [Ca2+]i transients were progressively smaller in amplitude, suggesting that [Ca2+]i released from intracellular stores helps to maintain a critical level of [Ca2+]i during physiologic firing activity. As the authors have recently reported, brief exposure to combined OGD induced a membrane hyperpolarization coupled to an increase in [Ca2+]i. In the presence of cyclopiazonic acid or ryanodine, the hyperpolarization and the rise in [Ca2+]i induced by OGD were consistently reduced. These data support the hypothesis that Ca2+ release from ryanodine sensitive Ca2+ pools is involved not only in the potentiation of the Ca2+ signals resulting from cell depolarization, but also in the amplification of the [Ca2+]i rise and of the concurrent membrane hyperpolarization observed in course of OGD in striatal large aspiny interneurons. PMID- 10826536 TI - High-resolution CMR(O2) mapping in rat cortex: a multiparametric approach to calibration of BOLD image contrast at 7 Tesla. AB - The blood oxygenation level-dependent (BOLD) functional magnetic resonance imaging (fMRI) method, which is sensitive to vascular paramagnetic deoxyhemoglobin, is dependent on regional values of cerebral metabolic rate of oxygen utilization (CMR(O2)), blood flow (CBF), and volume (CBV). Induced changes in deoxyhemoglobin function as an endogenous contrast agent, which in turn affects the transverse relaxation rates of tissue water that can be measured by gradient-echo and spin-echo sequences in BOLD fMRI. The purpose here was to define the quantitative relation between BOLD signal change and underlying physiologic parameters. To this end, magnetic resonance imaging and spectroscopy methods were used to measure CBF, CMR(O2), CBV, and relaxation rates (with gradient-echo and spin-echo sequences) at 7 Tesla in rat sensorimotor cortex, where cerebral activity was altered pharmacologically within the autoregulatory range. The changes in tissue transverse relaxation rates were negatively and linearly correlated with changes in CBF, CMR(O2), and CBV. The multiparametric measurements revealed that CBF and CMR(O2) are the dominant physiologic parameters that modulate the BOLD fMRI signal, where the ratios of (deltaCMR(O2)/CMR(O2)/(deltaCBF/ CBF) and (deltaCBV/CBV)/(deltaCBF/CBF) were 0.86 +/- 0.02 and 0.03 +/- 0.02, respectively. The calibrated BOLD signals (spatial resolution of 48 microL) from gradient-echo and spin-echo sequences were used to predict changes in CMR(O2) using measured changes in CBF, CBV, and transverse relaxation rates. The excellent agreement between measured and predicted values for changes in CMR(O2) provides experimental support of the current theory of the BOLD phenomenon. In gradient-echo sequences, BOLD contrast is affected by reversible processes such as static inhomogeneities and slow diffusion, whereas in spin-echo sequences these effects are refocused and are mainly altered by extravascular spin diffusion. This study provides steps by which multiparametric MRI measurements can be used to obtain high-spatial resolution CMR(O2) maps. PMID- 10826537 TI - Noninvasive assessment of the functional neovasculature in 9L-glioma growing in rat brain by dynamic 1H magnetic resonance imaging of gadolinium uptake. AB - Pathophysiologic parameters of the functional neovasculature and the blood-brain barrier of 9L-glioma in rat brain were measured noninvasively by dynamic 1H magnetic resonance imaging studies of gadolinium (Gd)-DTPA uptake. Changes of apparent [Gd-DTPA] uptake in time (CT[t]) were analyzed in a slice through the center of 10 9L-gliomas using fast T1 measurements. The distribution of the contrast agent was spatially correlated with the distribution of perfused microvessels as determined by immunohistochemical analysis. This method permits a distinction between perfused and nonperfused microvessels with a disrupted blood brain barrier. In transverse slices of the whole tumor, a spatial correlation was observed between CT maps and the two-dimensional distribution of perfused microvessels. In the next step, Gd-DTPA uptake rates were spatially related to the perfused microvessel density (Np) or vascular surface area (Sp). In tumor voxels with perfused microvessels, a linear correlation was found between Gd-DTPA uptake rate constants (k values) and Np or Sp. No correlation was observed between k values and the total microvessel density. These are the first data that show a relation between Gd-DTPA uptake rates and parameters of the functional neovasculature in 9L-glioma growing in rat brain. Now that Gd-DTPA uptake studies can be related to parameters of the functional neovasculature, they may be used more efficiently as a prognostic tool before or during therapy. PMID- 10826538 TI - Evidence for coupling between glucose metabolism and glutamate cycling using FDG PET and 1H magnetic resonance spectroscopy in patients with epilepsy. AB - The purpose of this study was to examine the relation between glucose metabolism and glutamate concentration in the human brain, in both the normal and diseased state. Regional values of glucose metabolism measured with 2-deoxy-2[F-18]fluoro D-glucose positron emission tomography (FDG PET) studies and single-voxel proton magnetic resonance spectroscopy (1H MRS) measurements of the glutamate/ glutamine/gamma-aminobutyric acid (Glx) tissue concentration were determined in multiple brain regions in 11 patients (5 girls and 6 boys, mean age 7.5 years) with medically intractable partial epilepsy. FDG PET and 1H MRS studies were performed in the interictal state in seven patients and in the ictal/periictal state in four patients. Regions of interest were identified in epileptic cortex (determined by intracranial and/or scalp electroencephalography) and in contralateral normal brain regions. Lower glucose metabolism and lower Glx concentrations were found in the epileptic focus than in the contralateral normal cortex in all seven patients examined in the interictal state, whereas higher glucose metabolism and higher Glx concentrations were observed in the epileptic focus in the four patients who had ictal/periictal studies. Significant correlations were found between the values of cerebral glucose utilization and Glx concentration in epileptic brain region, in nonepileptic brain regions, and in epileptic and nonepileptic regions combined. These results demonstrate a significant relation between glucose metabolism and glutamate/glutamine concentration in normal and epileptic cerebral cortex. This relation is maintained in both the interictal and ictal states. PMID- 10826539 TI - Modeling dynamic PET-SPECT studies in the wavelet domain. AB - This work develops a theoretical framework and corresponding algorithms for the modeling of dynamic PET-SPECT studies both in time and space. The problem of estimating the spatial dimension is solved by applying the wavelet transform to each scan of the dynamic sequence and then performing the kinetic modeling and statistical analysis in the wavelet domain. On reconstruction through the inverse wavelet transform, one obtains parametric images that are consistent estimates of the spatial patterns of the kinetic parameter of interest. The theoretical setup allows the use of linear techniques currently used in PET-SPECT for kinetic analysis. The method is applied to artificial and real data sets. The application to dynamic PET-SPECT studies was performed both for validation purposes, when the spatial patterns are known, and for illustration of the advantages offered by the technique in case of tracers with an unknown pattern of distribution. PMID- 10826540 TI - Ocular problems of young adults in rural Nigeria. AB - OBJECTIVES: To determine the common eye diseases as well as the prevalence and causes of blindness and visual impairment in young adult residents of rural areas of Anambra State, Nigeria. MATERIALS AND METHODS: Three rural villages in Anambra State, Nigeria were selected by simple random sampling. A structured questionnaire on demographic characteristics, symptoms and attitude to eye diseases was administered to residents aged 18-49 years. Ocular examination included visual acuity estimation, colour vision test, tonometry, refraction and ophthalmoscopy. Skin-snip was examined for microfilaria. RESULTS: The common ocular problems in the 510 young adults examined were presbyopia (33.3%), refractive errors (41.1%), allergic conjunctivitis (8.2%), pterygium (8.2%), pingueculum (5.9%) and colour vision defect (2.4%). Bilateral blindness occurred in 1.2%, uniocular blindness in 0.8% and 1.7% had visual impairment in their better eyes. Glaucoma and sequelae of congenital cataract caused bilateral blindness while visual impairment was due to refractive errors, cataract, corneal opacities and uveitis. Trauma predisposed to uniocular blindness and visual impairment. CONCLUSIONS: Eyeglasses alone would alleviate visual impairment and ensure good near vision in more than 47% of the subjects. The prevalence of blindness could be reduced through early detection of glaucoma, congenital cataract and ocular trauma. PMID- 10826541 TI - Solitary fibrous tumour of the orbit. Report of a new case. AB - PURPOSE: Solitary fibrous tumour (SFT) of the orbit is a very rare lesion that may be misdiagnosed as fibrous histiocytoma, haemangiopericytoma, or other orbital tumour. We studied a case of SFT of the orbit in a 35-year-old woman. METHODS: The patient, with a 1-year history, reported a 'pressure' sensation behind her right eye, but no pain, vision changes, or other associated symptoms. Ocular examination was normal. Magnetic resonance imaging (MRI) showed a well circumscribed medial mass in the right orbit. The tumour was excised. RESULTS: The tumour was a cellular spindle-cell neoplasm with a storiform pattern. The tumour cells were spindle-shaped with bland nuclei and rare mitoses. Immunostaining was positive for vimentin and CD34, but negative for cytokeratin (AE1-AE3), EMA, desmin, smooth muscle actin, S-100, CD31, CD45 and bcl-2. CONCLUSION: SFT can infrequently involve the orbit. The tumour must be removed entirely to avoid recurrences. Careful and continued follow-up is important because orbital recurrence may occur several years after the excision of the primary tumour. PMID- 10826542 TI - Childhood eye injuries in North Jordan. AB - AIMS: To provide epidemiological data on serious eye injuries among children that can be utilized in planning health education and safety strategies for preventing such injuries. MATERIALS AND METHODS: Records were reviewed of 116 children who sustained serious eye injuries which required admission to Princess Basma Teaching Hospital between October 1995 and November 1998. The material was analyzed retrospectively with respect to various epidemiological features. RESULTS: 71.5% of the injured children were male and 28.5% were female. There was a marked preponderance of injuries in the age group 6-10 years. The majority of injuries occurred during play and sport (74.1%). Stones and sharp objects were the most common causes accounting for 18.1% and 17.2% respectively. Most of the sharp objects were household instruments. Perforating injuries were more common than non-perforating injuries. 56% of injured eyes had a low vision with visual acuity between 6/24 and 3/60, and 13% had a blinding outcome with visual acuity less than 3/60. CONCLUSION: The major causes of serious eye injuries among these children were unnecessary risks during play, the use of dangerous objects and insufficient supervision by adults, especially at home. Health education should be directed towards prevention of such serious injuries. PMID- 10826543 TI - Intermediate uveitis of pars planitis type in identical twins. Report of a case. AB - The occurrence of intermediate uveitis in monozygotic twins has rarely been reported in literature. We report the occurrence of this disease condition in identical twins. Twin sisters presented with typical features of intermediate uveitis, which appeared three years earlier in one of the sisters. They were treated with posterior subtenon injection of triamcinolone acetonide and recovered well. Systemic evaluation and laboratory investigations did not reveal any etiology. Cytogenetic high-resolution analysis did not reveal any chromosomal abnormalities. Pedigree analysis revealed no specific transmission pattern. No other family members in three generations had similar ocular disease. Our report indicates that a detailed family history should be taken in intermediate uveitis and examination of other siblings of patients with intermediate uveitis should be performed. PMID- 10826544 TI - Retinopathy of prematurity: evaluation of risk factors. AB - OBJECTIVES: The purpose of this study was to determine what risk factors play a role in the development of retinopathy of prematurity (ROP). STUDY DESIGN: Data were collected on 157 infants born and cared for in one institution between January 1991 and July 1994. Initially we evaluated all children enrolled in the study to determine potential risk factors for the development of ROP. We subsequently compared multiple variables for ROP positive singletons with ROP positive twins to determine ROP risk factors for each group and to determine if one group was more susceptible to a given risk factor. RESULTS: Of the 157 infants examined, 72 infants (46%) developed ROP. Infants who developed ROP had a lower gestational age, a lower birth weight, a higher number of days on oxygen/ventilator, more days in the intensive care unit (ICU), a greater need for steroids and a higher incidence of sepsis when compared to infants who did not develop ROP. There was no significant difference noted between singleton and twin gestation infants that developed ROP when comparing gestational age, weight, ventilator time or length of ICU stay. Total number of days on oxygen therapy was higher in the singleton group and this difference did reach statistical significance. CONCLUSIONS: Several risk factors are associated with a higher incidence of ROP. These variables may not be independent risk factors but may be a sign of the increased severity of illness associated with those infants who are born earlier with a lower birth weight. Multiple gestational births do not appear to increase the risk of developing ROP when compared to a similar group of singleton birth infants. PMID- 10826545 TI - Effect of verapamil on lenticular calcium, magnesium and iron in radiation exposed rats. AB - PURPOSE: To investigate the effect of verapamil on lenticular calcium, magnesium, iron and on radiation-induced cataract in rats. METHODS: Thirty-seven, female, Wistar Albino rats, weighing 180-230 g were randomly grouped as follows: control group (10 rats), radiation group (13 rats) and radiation-verapamil group (14 rats). Both radiation and radiation-verapamil groups received 5 Gy radiation to the whole body in a single dose, including both eyes within the irradiation volume; in addition the verapamil group received daily subcutaneous injections of 8 mg/kg verapamil starting on the first day of radiation. At the end of an 8-week experimental period, the animals were killed by decapitation. Lenticular calcium, magnesium, and iron levels were studied. RESULTS: The mean lens calcium level for the radiation group was significantly higher than that for the control and radiation-verapamil groups and, there was no significant difference between the control and radiation-verapamil groups. The mean lens magnesium value for the radiation group was significantly higher than that for the control group. In the radiation-verapamil group the mean lens magnesium content was significantly lower than that for the radiation group. The iron level in the radiation group was significantly higher than that for the radiation-verapamil group. CONCLUSIONS: The lens calcium, magnesium and iron contents increased after radiation exposure. Verapamil treatment significantly reduced the increase in the lenticular content of calcium, magnesium and iron, indicating a probable protective effect of verapamil in radiation-induced cataract formation. PMID- 10826546 TI - Lactate dehydrogenase activity in pathological human tears obtained with glass capillaries correlates with the albumin content. AB - Lactate dehydrogenase (LDH) activity was measured in stimulated normal human tears and in tears from patients suffering from various corneal and conjunctival disorders. In normal tears the LDH activity was below the detection level of the method used, and it was hardly detectable in tears of most of the patients. The LDH activity correlated with the serum albumin content (r = 0.745, n = 32), suggesting that its source in pathological cases could be the plasma by transudation, and not the surface epithelial cells. Based on these results, LDH activity cannot be used as a measure of epithelial cell destruction in most pathological conditions, since the tear secretion of the patients is usually stimulated due to irritation, associated with the disorder or disease. PMID- 10826547 TI - Outdoor work and the risk of pterygia: a case-control study. AB - AIM: To investigate the relationship between outdoor work and pterygium formation in a hospital-based case-control study in Singapore. METHOD: Sixty-one patients with pterygium (49 males, mean age 54.2 years) and 125 controls (41 males, mean age 50.2 years), excluding patients with cataract and age-related macular degeneration presenting at outpatient clinics of the Singapore National Eye Centre were interviewed face-to-face with a standard questionnaire. Demographic data and information on outdoor work and eye protection, were collected. RESULTS: Thirty-eight subjects (20.4%) were outdoor workers and 148 (79.6%) were indoor workers. Outdoor workers were more likely to be male smokers and alcohol drinkers with lower family income and higher sunlight exposure than indoor workers. The crude odds ratio for pterygium in outdoor workers was 7.0 (95% confidence interval 3.2, 15.3). The adjusted odds ratio was 4.2 (95% confidence interval 1.7, 10.1). CONCLUSION: There is a statistically significant association between outdoor work and pterygium formation, which may be related to the higher exposure to sunlight and dust in outdoor workers. Efforts may be made to educate outdoor workers to wear proper equipment to protect their eyes against excessive sunlight. PMID- 10826548 TI - Ocular haemodynamics in glaucoma associated with high myopia. AB - The ocular blood flow of a group of high myopic and glaucomatous eyes has been investigated by means of the Color Doppler Imaging (CDI) technique, comparing the results with those of some very myopic eyes without glaucoma, some non-myopic glaucomatous eyes and with a control group. The Ophthalmic Artery (OA), the Short Posterior Ciliary Arteries (SPCAs) and the Central Retinal Artery (CRA) were considered. For every vessel, peak systolic velocity (SV), end-diastolic velocity (DV) and the Pourcelot Resistivity Index were studied. The statistical analysis was performed by means of t-test according to Bonferroni procedure for multiple comparisons. The authors registered in the group of eyes with glaucoma associated with very severe myopia some abnormalities of the OA blood flow systolic velocity which might be interpreted as a vascular risk factor for the pathogenesis of the glaucomatous optic neuropathy; these abnormalities are more severe in glaucoma patients with high myopia than in an age-matched group of glaucoma patients without myopia. PMID- 10826549 TI - Patient perceptions of quality-of-life associated with bilateral visual loss. AB - PURPOSE: To ascertain whether patients with unilateral visual loss to a specific level are able to approximate the degree of impairment of quality-of-life experienced by patients with bilateral visual loss to the same level. METHODS: One hundred thirty-three study group patients with (1) visual loss to 20/40 or worse in at least one eye, and (2) a marked difference between the visual acuities in their two eyes, were polled using the time tradeoff method of utility value measurement. All patients were asked to assume that the visual acuity in both of their eyes was as poor as the visual acuity in their worst seeing eye. These utility values were then compared to those obtained from a control group of 173 patients with known utility values who had similar bilateral visual loss. Both the study and control groups were stratified into 4 levels of visual loss (20/40 to 20/50, 20/60 to 20/100, 20/200 to 20/400, and counting fingers to light perception). RESULTS: Mean utility values for the study group ranged from 0.47 to 0.71. Patients with unilateral visual loss, given the assumption of bilateral visual loss to the same degree, routinely demonstrated no significant difference in utility preferences as compared to patients with true bilateral visual loss to the same level. CONCLUSIONS: Patients with unilateral visual loss can very accurately estimate the degree of impairment of quality-of-life that would result if visual loss to a similar degree occurred in the remaining eye with better vision. PMID- 10826550 TI - Visual field loss after argon laser panretinal photocoagulation in diabetic retinopathy: full- versus mild-scatter coagulation. AB - PURPOSE: The aim of the present study was to prospectively examine the effect of two styles of PRP, full- and mild-scatter on central and peripheral visual field in diabetic patients. METHODS: 40 eyes with preproliferative or early proliferative diabetic retinopathy of 32 patients underwent visual field testing on the C 30-1 and P 30/60-1 program of the Humphrey field analyser before and after argon laser panretinal photocoagulation (PRP). 21 eyes received full- and 19 mild-scatter PRP. RESULTS: Laser treatment had caused significant reduction of central retinal sensitivity in all eyes with no difference between full- and mild scatter PRP. Peripheral visual field sensitivity was significantly improved after mild-scatter PRP and was significantly depressed after full-scatter PRP. CONCLUSION: Our results suggest, that two treatment produced essentially equal effects on central and different on peripheral visual field. PMID- 10826551 TI - Guidelines for the use of self-blood pressure monitoring: a summary report of the First International Consensus Conference. Groupe Evaluation & Measure of the French Society of Hypertension. PMID- 10826552 TI - Renal proximal tubule sodium transport and genetic mechanisms of essential hypertension. AB - Renal sodium re-absorption is a closely regulated process serving to maintain both extracellular fluid volume and arterial blood pressure. Proteins participating in sodium re-absorption and its regulation are therefore important candidate proteins whose genes may contain sequence variation contributing to the inherited tendency for increased arterial blood pressure (essential hypertension). Important insight has come from rare forms of single-gene hypertension in human subjects and from polygenic animal models of genetic hypertension. Both indicate the primacy of altered renal function in the genesis of hypertension, and suggest that genes contributing to the disease are members of the subset of genes expressed in the kidney. This review examines evidence for abnormalities in renal sodium re-absorption in hypertension and focuses on the proximal tubule as a site of relevant dysfunction. Identification of the proteins participating in renal sodium re-absorption and its regulation, particularly those involved in the renal pressure-natriuresis mechanism, will allow gene cloning and sequencing which in turn may lead to the identification of novel gene sequence variation participating in hypertension. PMID- 10826553 TI - Hypertension in developing economies: a review of population-based studies carried out from 1980 to 1998. AB - AIM: Hypertension is an established major risk factor underlying the epidemic of coronary and cardiovascular diseases in most developed countries, and it has been shown to be a public health problem in many developing countries since the 1970s. This review was carried out to illustrate the current situation with regard to blood pressure levels and the prevalence and management of hypertension in developing countries. METHODS: A search for papers published in medical journals from 1991 to 1998 was performed using the MEDLINE database. A total of 40 articles were selected according to previously defined criteria. RESULTS: Hypertension is a public health problem for most of the developing countries reviewed, and it is frequently associated with low levels of awareness, treatment and control. A positive association between the gross national product per capita and the prevalence of hypertension in developing countries is also evident. CONCLUSIONS: Differences in methodology between national surveys make international comparisons difficult. Nevertheless, low-cost hypertension control programmes in developing countries are needed, along with development of the primary prevention of hypertension. PMID- 10826554 TI - The relationship of childhood to adult blood pressure: longitudinal study of juvenile hypertension in Lithuania. AB - OBJECTIVE: To assess changes in blood pressure from childhood to adulthood and the ability to predict adult blood pressure. DESIGN: Longitudinal study of a cohort of children with baseline data and a follow-up survey after 20 years. SETTING: Epidemiological survey of schoolchildren and subsequent inhabitants of Kaunas, a town in Lithuania. PARTICIPANTS: The children came from 15 schools and accounted for 25% of all 12- and 13-year-old children born in 1964 in Kaunas. The first survey (n = 1082) was carried out in 1977. The same population was re examined in 1997 (n = 505). Data from 217 men and 288 women, who participated in both the first and the most recent surveys, is presented. MAIN OUTCOME MEASURES: Systolic and diastolic blood pressure at the age of 32-33 years. RESULTS: In the 20 years between the two surveys blood pressure increased more in men than in women. Statistically significant correlation between childhood and adult blood pressure levels was estimated (for systolic blood pressure r=0.40 in men and r=0.24 in women; for diastolic blood pressure r=0.14 in men and r=0.34 in women). Stepwise regression analysis of the data showed that the best predictors of adult blood pressure were the initial childhood blood pressure levels and change in BMI during the 20-year period for both men and women. Other factors were less predictive. CONCLUSIONS: Childhood blood pressure is related to adult levels and, together with changes in body mass index, is a significant predictor of adult blood pressure. PMID- 10826555 TI - Influence of movement arousal on circadian rhythm of blood pressure in obstructive sleep apnea syndrome. AB - OBJECTIVE: To investigate the hypothesis that repeated arousals at the termination of apnea/hypopnea in obstructive sleep apnea syndrome (OSAS) are related to abnormal circadian rhythm of blood pressure (BP). DESIGN AND METHODS: We performed polysomnography (PSG) with pulse oximetry in 26 middle-aged patients with OSAS aged 42-58 years (mean age 51.8 years). The intensity of arousal on PSG was graded into two levels: grade 1 (EEG arousal, EA), an abrupt shift in EEG frequency, and grade 2 (movement arousal, MA), EEG arousal with an increase in electromyogram activity lasting at least 3 s. The number of apnea/hypopneas per hour (apnea/hypopnea index, AHI), and length of time during which nocturnal oxygen saturation decreased below 90% (oxygen desaturation time, ODT) were also evaluated. Percentage EA and %MA were assessed by the ratio of the number of apneas and hypopneas with EA or MA to the number of apneas and hypopneas in total. The 24 h systolic blood pressure (SBP) and diastolic blood pressure (DBP) were measured noninvasively. Multiple regression analysis was performed among AHI, ODT, %EA and %MA or among age, body mass index and %MA. RESULTS: The %MA was the most significant factor contributing to the elevated 24 h SBP (r= 0.46, P< 0.05); oxygen desaturation (r= 0.44, P< 0.05) was the next most important contributing factor. The level and pattern of 24 h BP differed significantly between the patients with %MA 85% and %MA < 85% (mean 24 h SBP: 147 +/- 16.8 versus 125 +/- 19.6 mmHg, P< 0.01; mean 24 h DBP: 97.5 +/- 14.3 versus 85.6 +/- 14.6 mmHg, P< 0.01), and also differed between those with severe OSAS, i.e. ODT > or = 130 min, and mild to moderate OSAS, i.e. ODT < 130 min, (mean 24 h SBP: 149 +/- 15.8 versus 132 +/- 20.6 mmHg, P < 0.01; mean 24 h DBP: 100 +/- 14.1 versus 87.4 +/- 14.0 mmHg, P < 0.01). CONCLUSION: Our findings suggest that MA and oxygen desaturation in OSAS make an important contribution to abnormal circadian rhythm of BP. We conclude that repeated end-apneic arousal and/or hypoxic asphyxia and the subsequent sleep fragmentation may contibute to nocturnal and diurnal elevation of BP. PMID- 10826556 TI - Mapping of quantitative trait loci (QTL) of differential stress gene expression in rat recombinant inbred strains. AB - OBJECTIVE: Stress has been shown to be a major environmental contributor to cardiovascular diseases through its effects on blood pressure variability and cardiac function. The cellular stress response is characterized by the expression of specific heat stress genes (hsps), under the transcriptional control of heat shock transcription factors (HSTFs). The levels of hsp mRNA depend on the severity of the stress, with hstf1 acting as a stress sensor. The aim of this work was to evaluate the genetic contribution of the variability in hsp expression, and to identify its putative quantitative trait loci (QTL). METHODS: Twenty recombinant inbred rat strains (RIS) were studied. The animals underwent a standardized, identical 1 h immobilization stress in restraint cages, followed by 1 h of rest before sacrifice. Total RNA was extracted from the heart kidneys and adrenals, and the mRNA levels of hsp27, hsp70, hsp84, hsp86 and hsp105 were measured. The strain distribution pattern (SDP) of hsp expression was correlated with that of 475 polymorphic markers distributed throughout the RIS genome. A polymorphism of rat hstf1 in RIS was used for its mapping in RIS. RESULTS: Despite an identical stress being applied to all strains, hsp expression showed up to a 1 2-fold gradient with little intra-strain variability, indicative of a strong genetic contribution to the trait Heritability ranged from 50 to 77% for most hsp genes in the three target organs. The continuous SDP of stress gene expression indicated the polygenic nature of the trait A common locus on chromosome 7 (at D7Cebrp187s3 marker) was consistently associated with all hsp expression in most of the organs [with a likelihood of odds (LOD) score of 3.0 for hsp27 expression]. We have mapped rat hstf1 on chromosome 7 at the same locus. Finally, the D4Mit19 marker was significantly associated with hsp84 expression in the heart (LOD score of 3.1). CONCLUSION: Two loci were linked with the differential expression of HSPs in response to immobilization stress in target organs of RIS. The chromosome 7 locus unveiled for all HSPs could explain up to 42% of the observed inter-strain variability of hsp levels in response to stress. We propose hstf1 as a positional candidate at this locus. PMID- 10826557 TI - Polymorphism in oestrogen response element associated with variation in plasma angiotensinogen concentrations in healthy pregnant women. AB - OBJECTIVE: To investigate the hypothesis that the genotype at nucleotide A(-20)C in the 5' flanking region of the angiotensinogen gene, which lies within a sequence with high homology to an oestrogen response element, affects plasma angiotensinogen levels in pregnancy. DESIGN: Prospective observational study METHODS: Seventy-two healthy pregnant women were recruited in the second half of pregnancy from hospital and primary care antenatal clinics in Nottingham, UK. Plasma angiotensinogen concentrations were measured by radioimmunoassay of angiotensin I generated from endogenous angiotensinogen in the presence of excess human renin. DNA was extracted from peripheral venous blood, and angiotensinogen genotype determined at A(-20)C, G(-6)A and Met235Thr. Associations between genotype and plasma angiotensinogen concentration were assessed by analysis of variance. RESULTS: Women homozygous for the -20C allele had the lowest mean plasma angiotensinogen concentration of 1.7 +/- 0.3micromol/l. Women homozygous for -20A had significantly higher plasma angiotensinogen concentrations (2.6 +/- 0.1 micromol/l), and intermediate levels (2.0 +/- 0.1 micromol/l) were observed in women heterozygous for A(-20)C (P = 0.002, ANOVA). The polymorphisms at nucleotide -6 and codon 235 were in almost complete linkage disequilibrium, and nucleotide -20C was found only in a subset of -6A/235Thr alleles. Conclusion The low plasma angiotensinogen levels associated with the -20C/-6A/235Thr haplotype in pregnant women contrast with the high concentrations associated with the 235Thr allele in the non-pregnant state. A possible explanation lies in the presence of a motif with high homology to an oestrogen response element between the TATA box and transcription initiation site. Previous in vitro studies of reporter gene constructs have demonstrated thatthe A(-20)C polymorphism affects oestrogen responsiveness. The results of this study support the hypothesis that the oestrogen response element of the angiotensinogen gene is of functional importance in pregnancy, and that oestrogen responsiveness in pregnancy is influenced by the genotype at nucleotide - 20. PMID- 10826558 TI - Alterations in skeletal muscle morphology in glucose-tolerant elderly hypertensive men: relationship to development of hypertension and heart rate. AB - OBJECTIVE: To compare the muscle morphology in hypertensive subjects with that in controls and to test the hypothesis of a relation between heart rate, development of hypertension and muscle morphology that is independent of glucose intolerance. PATIENTS AND METHODS: We studied 43 glucose-tolerant, untreated hypertensive subjects and 113 healthy controls in a longitudinal cohort of 70-year-old men. Metabolic status (oral glucose tolerance test and euglycemic, hyperinsulinaemic clamp test), muscle fibre distribution (myosin ATPase staining) and capillary supply (amylase-PAS method) were evaluated. Blood pressure and heart rate data were available from both ages 50 and 70 years. RESULTS: Hypertensive subjects had a significantly smaller mean number of capillaries per fibre than controls (1.53 versus 1.64; P = 0.04). In hypertensive subjects, the proportions of type I and type II fibres were correlated to mean arterial pressure (r = -0.56 and r= 0.52, respectively, P < 0.05 for both). The increase in mean arterial pressure over 20 years was closely correlated to capillary density in mm2 (r= -0.62; P< 0.0001). Capillary supply was inversely related to resting heart rate both at ages 50 and 70 years. CONCLUSIONS: Skeletal muscle of glucose tolerant hypertensive subjects showed a lower capillary supply than that of controls. This capillary rarefaction was correlated to increase in mean arterial pressure over two decades and to supine heart rate. This is compatible with the suggestion that higher sympathetic drive might generate structural alterations in muscle capillarization. PMID- 10826559 TI - Dietary alpha-lipoic acid supplementation lowers blood pressure in spontaneously hypertensive rats. AB - BACKGROUND AND OBJECTIVES: In spontaneously hypertensive rats (SHRs), excess endogenous aldehydes bind sulfhydryl groups of membrane proteins, altering membrane Ca2+ channels and increasing cytosolic free calcium and blood pressure. The thiol compound, N-acetyl cysteine, normalizes elevated blood pressure in SHRs by binding excess endogenous aldehydes and normalizing membrane Ca2+ channels and cytosolic free calcium. The aim of the present study was to investigate whether a dietary supplementation of an endogenous fatty acid, alpha-lipoic acid, another thiol compound that is known to increase tissue cysteine and glutathione, can lower blood pressure and normalize associated biochemical and histopathological changes in SHRs. METHODS AND RESULTS: Starting at 12 weeks of age, animals were divided into three groups of six animals each. Animals in the Wistar- Kyoto (WKY) rat control group and the SHR control group were given a normal diet, and the SHR lipoic acid group was given a diet supplemented with lipoic acid (500 mg/kg feed) for the next 9 weeks. After 9 weeks, systolic blood pressure, platelet [Ca2+]i, plasma insulin and liver, kidney and aortic aldehyde conjugates were significantly higher in SHR controls as compared with WKY rat controls and the SHR lipoic acid group. SHR controls also showed smooth muscle cell hyperplasia in the small arteries and arterioles of the kidneys. CONCLUSIONS: Dietary alpha lipoic acid supplementation in SHRs lowered the systolic blood pressure, cytosolic [Ca2+]i, blood glucose and insulin levels, and tissue aldehyde conjugates, and attenuated adverse renal vascular changes. PMID- 10826560 TI - Oxidative stress augments secretion of endothelium-derived relaxing peptides, C type natriuretic peptide and adrenomedullin. AB - OBJECTIVE: Excess oxidative stress is one of the major metabolic abnormalities on vascular walls in hypertension and atherosclerosis. In order to further elucidate the endothelial function under oxidative stress, the effect of hydrogen peroxide (H2O2) on expression of two novel endothelium-derived vasorelaxing peptides, C type natriuretic peptide (CNP) and adrenomedullin (AM) from bovine carotid artery endothelial cells (BCAECs) was examined. METHODS: BCAECs were treated with H2O2 (0.1-1.0 mmol/ l) and/or an antioxidant, N-acetylcysteine (NAC) (5-10 mmol/l), and incubated for 48 h. The concentrations of CNP and AM were measured with the specific radioimmuno assays that we originally developed. CNP and AM mRNA expressions were also examined by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: Treatment of BCAECs with 0.5 and 1 mmol/l H2O2 induced 9-and 10-fold increases of CNP concentration in the media. Addition of 10 mmol/l NAC significantly suppressed the effect of H2O2 by 52%. RT-PCR analysis showed that CNP mRNA expression in BCAECs was also rapidly augmented within 1 h with H2O2 (1 mmol/l) treatment, and reached a peak at 3 h to show a 10-fold increase. AM secretion from BCAECs also increased to two-fold with exposure to 0.5 mmol/l H2O2, accompanied with the augmented level of AM mRNA. NAC 10 mmol/l completely suppressed the effect of H2O2 on AM secretion. CONCLUSIONS: In this study, it has been demonstrated that H2O2 augments endothelial secretion of the two endothelium derived relaxing peptides, CNP and AM. Our findings suggest the increased secretion of CNP and AM from endothelium under oxidative stress may function to compensate the impaired nitric oxide-dependent vasorelaxation in hypertension and atherosclerosis. PMID- 10826561 TI - Persistent high job demands and reactivity to mental stress predict future ambulatory blood pressure. AB - OBJECTIVE: To test the hypothesis that work stress (persistent high job demands over 1 year) in combination with high reactivity to mental stress predict ambulatory blood pressure. DESIGN: Assessment of cardiovascular responses to standardized behavioural tasks, job demands, and ambulatory blood pressure over a working day and evening after 12 months. PARTICIPANTS: We studied 81 school teachers (26 men, 55 women), 36 of whom experienced persistent high job demands over 1 year, while 45 reported lower job demands. METHODS: Participants were divided on the basis of high and low job demands, and high and low systolic pressure reactions to an uncontrollable stress task. Blood pressure and concurrent physical activity were monitored using ambulatory apparatus from 0900 to 2230 h on a working day. RESULTS: Cardiovascular stress reactivity was associated with waist/hip ratio. Systolic and diastolic pressure during the working day were greater in high job demand participants who were stress reactive than in other groups, after adjustment for age, baseline blood pressure, body mass index and negative affectivity. The difference was not accounted for by variations in physical activity. CONCLUSIONS: Cardiovascular stress reactivity and sustained psychosocial stress may act in concert to increase cardiovascular risk in susceptible individuals. PMID- 10826562 TI - Sympathetic and reflex alterations in systo-diastolic and systolic hypertension of the elderly. AB - BACKGROUND: Previous studies have shown that young and middle-aged essential hypertensives are characterized by a sympathetic activation coupled with an impaired baroreflex-heart rate control. The present study aimed to determine whether these neuroadrenergic and reflex alterations also characterize systo diastolic and systolic hypertension of the elderly. SUBJECTS AND METHODS: In 20 untreated elderly essential hypertensive subjects [10 with a systo-diastolic and 10 with an isolated systolic hypertension, aged 67.2 +/- 1.5 years and 66.9 +/- 1.7 years (mean +/- SEM)], we measured beat-to-beat arterial blood pressure (finger photoplethysmographic device), heart rate (electrocardiogram) and efferent postganglionic muscle sympathetic nerve activity (microneurography) at rest and during baroreceptor stimulation and deactivation induced by stepwise intravenous infusions of phenylephrine and nitroprusside, respectively. Data were compared with those obtained in 11 age-matched normotensive control subjects. RESULTS: Compared to the elderly normotensive group, muscle sympathetic nerve activity was increased to a similar degree in the group of systo-diastolic and systolic hypertension (50.8 +/- 4.2 versus 75.2 +/- 5.2 and 70.4 +/- 5.1 bursts per 100 heart beats, respectively, P< 0.01 for both). In the control group, the stepwise increase in arterial pressure induced by phenylephrine caused progressive bradycardia and sympathoinhibition, while the stepwise decrease in arterial pressure had opposite effects. While baroreceptor-heart rate control was markedly impaired (average reduction 41.6%), in both systo-diastolic and systolic hypertensive patients, baroreceptor modulation of sympathetic nerve traffic was similar to that seen in normotensive individuals. CONCLUSIONS: These data demonstrate that sympathetic activation is not only a feature of young and middle aged, but also of elderly hypertensives, regardless of whether both systolic and diastolic or only systolic blood pressure is increased. They also show that hypertension of the elderly is not accompanied by an impaired baroreceptor modulation of sympathetic nerve traffic. PMID- 10826563 TI - Reflex cardiovascular responses to somatic stimulation in spontaneously hypertensive rats. AB - OBJECTIVES: This study aimed to test whether the cardiovascular responses to somatic stimulation in spontaneously hypertensive rats (SHR) were enhanced compared with those in normotensive Wistar-Kyoto (WKY) rats, and to examine any role of the impaired baroreflex function in the hypertensive rats. METHODS: Experiments were done in anaesthetized SHR (n = 34) and WKY (n = 31). Baroreceptor reflexes were assessed by continuous infusion of incremental doses (5-30 microg/kg per min) of phenylephrine over a 3 min infusion period. Cardiovascular responses to sciatic nerve stimulation (5 s trains, 1 ms pulse duration, 400 microA intensity) were studied before and after baroreceptor deactivation. The latter was achieved either by carotid occlusion and cutting the vagi and aortic nerves (SHR, n = 28 and WKY rats, n = 27), or by complete baroreceptor denervation (SHR, n = 6 and WKY rats, n = 4). RESULTS: We confirmed that baroreceptor sensitivity was significantly lower in SHR (0.40 +/- 0.05 ms/mmHg) than in WKY rats (0.90 +/- 0.04 ms/mmHg). Sciatic nerve stimulation elicited significantly greater increases in mean arterial pressure (MAP) and in heart rate in SHR than in WKY rats (+32.5 +/- 1.9 mmHg versus +20.2 +/- 1.1 mmHg and +13.5 +/- 1.5 bpm versus +8.0 +/- 1.1 bpm, respectively). Following baroreceptor deactivation, the responses to the same sciatic nerve stimulation of MAP and heart rate in SHR (+38.5 +/- 2.4 mmHg and +15.5 +/- 1.5 bpm) were still significantly greater than those in WKY rats (+29.5 +/- 1.3 mmHg and +11.6 +/- 1.2 bpm). CONCLUSIONS: These results show that cardiovascular responses to sciatic nerve stimulation are increased in SHR compared to WKY rats, and that this increased reactivity to somatic stimuli in hypertensive rats does not depend upon the impairment in baroreflex function demonstrated in this strain. PMID- 10826564 TI - Nitric oxide modulates the development and surgical reversal of renovascular hypertension in rats. AB - OBJECTIVE: To evaluate the role of nitric oxide (NO) in the development and unclipping-induced reversal of blood pressure and bilateral renal function in two kidney, one clip (2K1C) Goldblatt hypertensive rats. METHODS: Goldblatt hypertensive rats were prepared by clipping the left renal artery 4 weeks before unclipping experiments. NG-nitro-L-arginine methyl ester (L-NAME) was administered after clipping and during unclipping to inhibit nitric oxide (NO) synthesis. Blood pressure and bilateral renal responses were measured. RESULTS: Chronic L-NAME treatment accelerated and aggravated blood pressure elevations and increased plasma nitrite and nitrate levels in 2K1C rats. Surgical removal of the renal artery clip induced profound reductions in blood pressure in rats with and without L-NAME treatment. However, the magnitude of the unclipping-induced depressor response at the first post-unclipping hour was significantly smaller in L-NAME-treated rats compared to those without L-NAME administration (15 +/- 1 versus 22 +/- 1%, P < 0.05). Two hours after unclipping, blood pressure of both groups fell to a comparable, normal level. Acute intravenous infusion of L-NAME in established 2K1C hypertensive rats further increased blood pressure. Subsequent unclipping caused a depressor response similar to that observed in hypertensive rats treated chronically with L-NAME. Despite the marked decreases in blood pressure, unclipping induced striking increases in glomerular filtration rate (GFR), urine flow and sodium and potassium excretion rates in the ipsilateral kidney. However, the magnitudes of increases in GFR and the diuretic and natriuretic responses in rats without L-NAME treatment were significantly greater than in rats with L-NAME administration. In contrast, unclipping reduced these function indices in the contralateral kidney to a similar level in rats with and without L-NAME treatment. CONCLUSIONS: NO exerts vasodilator action and thereby lessens renal artery clipping-induced blood pressure elevation. Furthermore, unclipping-induced release of NO partially contributes to the early reduction in blood pressure and changes in bilateral renal function but does not directly mediate the normalization of blood pressure after unclipping in this hypertension model. PMID- 10826565 TI - Effects of sodium intake on the pressor and renal responses to nitric oxide synthesis inhibition in normotensive individuals with different sodium sensitivity. AB - OBJECTIVE: The present study evaluated the role of nitric oxide (NO) in the systemic vascular and renal adaptation to changes in dietary sodium intake. DESIGN AND METHODS: Seven healthy normotensive male subjects were randomized to high or low sodium diets in a double blind crossover design (7 days on each diet). The NO synthesis inhibitor, NGmonomethyl-L-arginine (L-NMMA) was infused systemically (1.8 mg/kg over 30 min) at the end of each dietary period and its effects on blood pressure, renal plasma flow, glomerular filtration rate, urinary flow rate and sodium excretion were measured. RESULTS: Blood pressure increased in response to L-NMMA on a high sodium diet only (area under time curve percentage change in mean blood pressure, low sodium = -94.5 +/- 164.3; high sodium = 391.1 +/- 228.6; P < 0.05 low versus high). The increase in blood pressure was directly and significantly associated with the individual salt sensitivity, defined by the difference in systemic mean blood pressure between high and low sodium diets (r = 0.756; P < 0.05). L-NMMA also reduced renal plasma flow and urinary flow rate in subjects on high sodium diet. CONCLUSIONS: The data support a significant influence of endogenous NO in the systemic and renal vascular adaptation to a high sodium diet in normotensive men. In addition, the direct association between the individual sodium-sensitivity and the pressor response to L-NMMA suggests that there is increased dependence of vascular tone on NO in normotensive subjects whose blood pressure is more sodium sensitive. PMID- 10826566 TI - Increased proximal tubule NHE-3 and H+-ATPase activities in spontaneously hypertensive rats. AB - OBJECTIVE: To investigate renal proximal tubular sodium-hydrogen exchanger 3 (NHE 3) and H+-ATPase activities in young (5-week-old) spontaneously hypertensive rats (SHR) and normotensive Donryu (DRY) rats, in the period during which high blood pressure is developing. METHODS: Five-week-old SHR and DRY rats were weighed and systolic blood pressure recorded. Proximal tubule cells were isolated, loaded with the intracellular pH dye, 2'-7'-bis-carboxyethyl-5(6)-carboxyfluorescein acetoxymethyl-ester and acidified with a NH4+/NH3 prepulse. Na+-independent intracellular pH recovery rate (H+-ATPase activity) and initial Na+-dependent intracellular pH recovery rate (NHE-3 activity) were assessed. NHE-3 activity was assessed during inhibition of H+-ATPase with Bafilomycin A1 and during inhibition of any possible NHE-1 activity with Hoe 694. RESULTS: Mean body weight and systolic blood pressures of 5-week-old SHR and DRY rats were not significantly different. NHE-3 activity was higher in SHR, 1.08 +/- 0.1 pH units/min compared with DRY rats, 0.73 +/- 0.1 pH units/min (P < 0.05) H+-ATPase activity was also higher in SHR, 0.119 +/- 0.02 pH units/min, compared with DRY rats, 0.051 +/- 0.02 pH units/min (P < 0.05). CONCLUSIONS: Proximal tubule cells of 5-week-old SHR have higher NHE-3 and H+-ATPase activities compared with age-matched DRY rats. Enhanced proximal tubular fluid reabsorption is likely to contribute to development of high blood pressure in young SHR. PMID- 10826567 TI - Influence of gender and age on preventing cardiovascular disease by antihypertensive treatment and acetylsalicylic acid. The HOT study. Hypertension Optimal Treatment. AB - OBJECTIVE: We have assessed the influence of gender and age on the main outcome results of the Hypertension Optimal Treatment (HOT) study. DESIGN AND INTERVENTIONS: The aims of the HOT study were to study the relationship between three levels of target office diastolic blood pressure (BP) (< or = 90, < or = 85 or < or = 80 mmHg) and cardiovascular (CV) events in hypertensive patients, and to examine the effects of 75 mg acetylsalicylic acid (ASA) daily versus placebo. SETTING: Outpatient clinical trial in 26 countries. PATIENTS: A total of 18790 patients (mean age 61.5 years, range 50-80) were randomized and followed for an average of 3.8 years until 71051 patient-years and 683 events had occurred. MAIN OUTCOME MEASURES: CV death, myocardial infarction (MI) and stroke. RESULTS: There were significantly fewer MIs in those in the lower diastolic BP target groups (3.0 versus 1.2 and 1.7 MIs/1000 patient-years, P for trend = 0.034) in women (n = 8883), whereas the similar but smaller trend (4.1 versus 4.1 and 3.4 MIs/1000 patient-years) was not statistically significant in men nor in the subgroup analysis of younger and older subjects. The effect of ASA on preventing MI was not influenced by age < 65 years (P= 0.02) or age > or = 65 years (P = 0.04) but was influenced by gender (P = 0.38 in women and P = 0.001 in men, lowered by 42% corresponding to a reduction from 5.0 to 2.9 MIs/1000 patient-years). CONCLUSIONS: The data of this HOT study sub-analysis suggest somewhat differentiated optimal gender- and age-dependent effects of anti-hypertensive and anti-platelet therapies; lowering of diastolic BP to about 80 mmHg in hypertensive women and, in addition, the administration of 75 mg of ASA to well treated hypertensive men appear to effectively reduce the most common cardiovascular complication, i.e. myocardial infarction, in patients with essential hypertension. PMID- 10826568 TI - Reduced intestinal epithelial cell brush border membrane calcium transport in spontaneously hypertensive rats. PMID- 10826569 TI - The National Sentinel Caesarean Section Audit. PMID- 10826570 TI - Nitric oxide in parturition. PMID- 10826571 TI - Inter-observer variation in histopathological diagnosis and grading of vulvar intraepithelial neoplasia: results of an European collaborative study. AB - OBJECTIVE: To evaluate the inter-observer variability of vulvar intraepithelial neoplasia diagnosis and grading system. DESIGN: Prospective study. SAMPLE: Histological sections of 66 vulvar biopsies. METHODS: Six consultant pathologists working at different European institutions independently reviewed 66 vulvar biopsies. The following variables were investigated: specimen adequacy, gross categorisation into benign or neoplastic changes, presence of atypical cytological pattern, presence of neoplastic architectural pattern, grade of vulvar intraepithelial neoplasia, presence of histopathologic associated findings for human papillomavirus infection. MAIN OUTCOME MEASURES: The degree of inter observer variation for each histopathologic parameter was assessed by Kappa (kappa) statistics. The frequency and the degree of disagreement were calculated by a symmetrical agreement matrix showing the number paired classifications. RESULTS: A good agreement (overall weighted kappa = 0.65, unweighted kappa = 0.46) was observed for grading vulvar intraepithelial neoplasia. Human papillomavirus infection associated findings and specimen adequacy were the variables with less inter-observer agreement (overall weighted kappa 0.26 and 0.22, respectively). Exact agreement between two pathologists for grade of vulvar intraepithelial neoplasia was observed in 63.6% of paired readings; the rate of paired agreement reached 73.9% considering vulvar intraepithelial neoplasia 2 and 3 as a single class. Conversely, only 5.0% of vulvar intraepithelial neoplasia 1 diagnoses were concordant in paired analysis. CONCLUSIONS: Current terminology offers a reproducible tool in the hands of expert pathologists. While on the diagnosis of 'high grade' vulvar intraepithelial neoplasia (vulvar intraepithelial neoplasia 2 and 3) there is good agreement, the diagnostic category of vulvar intraepithelial neoplasia 1 is not reproducible. PMID- 10826572 TI - High risk human papillomavirus in women with normal cervical cytology prior to the development of abnormal cytology and colposcopy. AB - OBJECTIVE: To study the significance of the presence of high risk human papillomavirus (HPV) in women with initially normal cervical cytology for the development of abnormal cytology and an abnormal colposcopic impression. DESIGN: Prospective, observational study PARTICIPANTS AND METHODS: Sixty-eight women with cytomorphologically normal smears and at least one positive HPV test result were evaluated every six months by cytology, colposcopy and HPV testing. The endpoint of the study was abnormal cervical cytology. RESULTS: The median time of follow up from the first positive HPV test was 34 months. A total of 17 women developed abnormal cytology, of whom 16 (94%) had persistence of a high risk HPV infection. Women with persistent high risk HPV were more likely to develop abnormal cervical cytology than women without high risk HPV (hazard ratio 28.2, 95% CI 3.72-215.2); they also had an increased risk of developing an abnormal colposcopic impression (hazard ratio 4.4, 95% CI 1.69-11.7). Among the 17 women with abnormal cytology, high grade dysplasia was histopathologically demonstrated in eight women. CONCLUSION: Persistent presence of high risk HPV in normal cervical smears is associated with a significantly increased risk of developing abnormal cytology and to a lesser degree with developing an abnormal colposcopic impression. PMID- 10826573 TI - A clinical review of borderline glandular cells on cervical cytology. AB - OBJECTIVE: To review the diagnoses and diagnostic pathway of women presenting with borderline glandular cells on cervical cytology. To outline the basis of clinical approach of these women. DESIGN: Retrospective review. POPULATION: Forty three women referred to the hospital department over a 32-month period. METHODS: Review of the casenotes for the demographic data, previous cervical cytology and/or histology report, indication for the smear resulting in borderline glandular cells, colposcopic findings, diagnostic and/or treatment procedures, final diagnosis and current status. RESULTS: The average age was 36.7 years. Twenty-four women (56%) had clinically significant lesions: seven women (16%) presented with cancers, of which one was endometrial in origin, and 17 (40%) with intraepithelial neoplasia (CIN and cervical glandular intraepithelial neoplasia (CGIN)). Sixty-seven percent of all clinically significant lesions were of squamous origin. Thirty-seven had histological diagnosis, while six went on to cytological surveillance. Colposcopy was the most significant predictor for clinically significant lesions (P < 0.05). Punch biopsies and loop excisions were diagnostic when based on abnormal colposcopic findings. Brush cytology was appropriate follow up for asymptomatic, premenopausal women with no colposcopic abnormality. In addition, endometrial sampling was recommended in the peri- and postmenopausal women. CONCLUSION: Borderline glandular cells have a high incidence of clinically significant lesions. Immediate referral for colposcopy and assessment is strongly recommended in women with two borderline glandular smears to avoid delays in potential cancer diagnosis. PMID- 10826574 TI - A randomised double-blind placebo-controlled trial of transcervical intrauterine local anaesthesia in outpatient hysteroscopy. AB - OBJECTIVE: To assess whether transcervical intrauterine instillation of local anaesthetic agent reduces pain during diagnostic outpatient hysteroscopy and endometrial biopsy. DESIGN: Prospective, randomised, double blind, placebo controlled trial. SETTING: Outpatient hysteroscopy clinic in a university teaching hospital. POPULATION: Ninety women undergoing outpatient diagnostic hysteroscopy with or without endometrial biopsy. METHODS: Transcervical intrauterine instillation of 5 mL of 2% lignocaine into the uterine cavity before performing the procedure. MAIN OUTCOME MEASURES: Evaluation of pain at different stages of the procedure using a visual analogue scale and changes in blood pressure and heart rate. RESULTS: The use of local anaesthetic did not alleviate pain experienced during hysteroscopy and endometrial biopsy. It did not prevent the occurrence of vaso-vagal reactions; however the incidence of these was low. CONCLUSIONS: Transcervical instillation of local anaesthesia neither reduced pain nor prevented vaso-vagal reaction during hysteroscopy and endometrial biopsy. PMID- 10826575 TI - Effect of SR49059, an orally active V1a vasopressin receptor antagonist, in the prevention of dysmenorrhoea. AB - OBJECTIVE: To investigate the clinical effect of SR49059 when given shortly before the onset of menstruation as a preventative treatment of dysmenorrhoea. DESIGN: A double-blind, randomised, placebo-controlled, cross-over trial in complete block design (three periods, three treatments). SETTING: A clinical research organisation in Paris, France. PARTICIPANTS: Women aged 18-35 years suffering from primary dysmenorrhoea. INTERVENTIONS: In each of three menstrual cycles, women reported to the study centre and were given a daily dose of either placebo, 100 mg or 300 mg SR49059 from a minimum of 4 hours up to a maximum of three days before the onset of bleeding and/or menstrual pain. If this did not control the pain, women were allowed once a day to take a second dose of study treatment providing that at least 4 hours had passed since the first drug intake. MAIN OUTCOME MEASURES: Intensity of menstrual pain recorded by means of a visual analogue scale. Rating of symptoms of dysmenorrhoea (mainly back and pelvic pain) in relation to functional capacity (Sultan score). Self-assessment of menstrual blood loss in a menstrual diary record. RESULTS: Analysis of intensity of menstrual pain, as recorded by visual analogue scale and Sultan pain score (back and pelvic pain) during the first 24 hours of dysmenorrhoea, showed a dose related effect of SR49059. The 300 mg dose of SR49059 was significantly more effective than placebo. Similarly, a dose-related effect of SR49059 was shown on total Sultan score. SR49059 was well tolerated and no significant effect on the bleeding pattern was noted. CONCLUSIONS: This study showed for the first time a therapeutic effect of an orally active vasopressin V1a receptor antagonist in the prevention of dysmenorrhoea. Further studies are required to examine effect mechanisms and determine effective doses. PMID- 10826576 TI - Identifying the indications for laparoscopically assisted vaginal hysterectomy: a prospective, randomised comparison with abdominal hysterectomy in patients with symptomatic uterine fibroids. AB - OBJECTIVE: To compare laparoscopically assisted vaginal hysterectomy (LAVH) and total abdominal hysterectomy (TAH) in patients with uterine fibroids. DESIGN: A prospective randomised study. SETTING: The San Paolo Hospital, Milan. POPULATION: Sixty-two patients, who were not suitable for a vaginal hysterectomy, requiring treatment for uterine fibroids. METHODS: Randomisation between LAVH and TAH. Comparison of outcomes on the whole series, patients with uteri < or = 500 g (Group 1) and patients with uteri > 500 g (Group 2). MAIN OUTCOME MEASURES: To establish operating time, blood loss, complications, febrile morbidity, analgesics administration and hospital stay for both treatment approaches. RESULTS: Median uterine weight was 400 g in both LAVH and TAH group. Median operating time was longer for LAVH (135 min compared with 120 min for TAH; P = 0.001), but patients undergoing LAVH had less analgesics administration (23% compared with 77%, P < 0.001) and a shorter median hospital stay (3.8 compared with 5.8 days; P < 0.001). LAVH, when compared with TAH in the two weight subgroups, required a significantly longer operating time only in Group 2, significantly reduced analgesics administration only in Group 1, and significantly reduced hospital stay in both groups. Conversions of LAVH to laparotomy were significantly more frequent in Group 2 (3/11) than in Group 1 (0/20) (P = 0.04). CONCLUSIONS: Compared with TAH, LAVH has advantages in removing uteri weighing < or = 500 g, with comparable operating time, less post operative pain and shorter recovery. Among uteri weighing > 500 g LAVH showed a shorter recovery, but longer operating time than TAH and a 27% rate of conversion to laparotomy. PMID- 10826577 TI - Fertility outcome following combined methotrexate treatment of unruptured extrauterine pregnancy. AB - OBJECTIVE: To assess the effectiveness of systemic treatment with methotrexate in combination with local injection for unruptured tubal pregnancy, and to evaluate reproductive function following treatment. DESIGN: Prospective, open clinical study. SETTING: University clinic. POPULATION: Sixty-seven women with unruptured tubal pregnancy. METHODS: Systemic methotrexate (intramuscular methotrexate 0.5 mg/kg for up to five days) in combination with local application of 12.5 mg methotrexate via laparoscopy. MAIN OUTCOME MEASURES: The subsequent surgical intervention required and future fertility. RESULTS: In 89.6% of women no further surgical intervention was required and 47 women (81%) experienced subsequently an intrauterine pregnancy. In 39 of 40 women who underwent hysterosalpingography following treatment, patency of the affected tube was observed. CONCLUSIONS: Combined local and systemic methotrexate treatment for unruptured tubal pregnancy seems to be more effective than each therapeutic modality alone. PMID- 10826578 TI - Reproductive failure in women living in iodine deficient areas of West Africa. AB - OBJECTIVES: To investigate the association between iodine status and reproductive failure in a population of West African women. DESIGN: Epidemiological survey on iodine deficiency disorders carried out in 1996-1997. SETTING: The iodine deficient areas of Senegal (Casamance and Senegal Oriental). POPULATION: Four thousand nine hundred and eighty women, aged 10 to 50, of whom 1,544 adolescent and 462 pregnant women were examined for thyroid size and urinary iodine excretion. Their iodine status was associated with their fertility rate and reproductive failures. RESULTS: Reproductive failure (defined as repeated miscarriages and stillbirth) was associated with low iodine status, with severe iodine deficiency increasing the risk. Poor nutritional status and illiteracy had a significant effect on the outcome of pregnancy: underweight women had a fourfold higher risk, and those who were illiterate an eightfold higher risk, of failed pregnancy, compared with nutritionally healthy, literature women. CONCLUSIONS: These findings emphasise the need to implement an effective iodine supplementation programme targeted at young and pregnant women in this area of western Africa. They also emphasise the importance of improving the nutritional status of young girls and the crucial role played by education in the prevention of reproductive failure. PMID- 10826579 TI - An epidemic of parvovirus B19 in a population of 3,596 pregnant women: a study of sociodemographic and medical risk factors. AB - OBJECTIVES: To estimate the incidence of human parvovirus B19 among pregnant women before and during an epidemic, to elucidate possible sociodemographic and medical risk factors during pregnancy and to estimate the association between parvovirus B19 infection and negative pregnancy outcome. DESIGN: Prospective study among pregnant women followed from their first antenatal visit before 24 full weeks of gestation until delivery. SETTING: Department of Obstetrics and Gynaecology, Odense University Hospital, Denmark, November 1992 to February 1994. METHODS: 3,596 pregnant women were invited to participate. The women were examined at first antenatal visit in the period from November 1992 to February 1994 and at delivery. The last delivery was in August 1994 and samples were thus collected before and during a large parvovirus B19 epidemic in Denmark January to September 1994. A blood sample for parvovirus B19 serology was taken at enrollment and from the umbilical cord at delivery. Three questionnaires were completed during 2nd and 3rd trimesters and a registration form at delivery. In total, 3,174 (87.6%) were enrolled and 79.5% completed the study. RESULTS: The prevalence of B19 IgG seropositivity at the first antenatal visit before 24 full weeks of gestation was 66% . The cumulative prevalence proportion of acute parvovirus B19 infection during pregnancy among IgG negative women was found to be 10.3% (IgM seropositivity and/or IgG seroconversion). The IgG seroconversion incidence increased significantly from 1.0% to 13.5% among 932 seronegative pregnant women before and during the epidemic, respectively (P < 0.001). Independent risk factors related to increased risk of B19 infection during pregnancy, adjusted for other sociodemographic and medical factors, were: children at home (adjusted OR 2.1, 95% CI 1.3-3.2); serious medical disease (adjusted OR 3.0, 95% CI 1.0-8.5); and a stressful job (adjusted OR 1.8, 95% CI 1.0-3.3). Parvovirus B 19 IgM seropositivity was associated with events of late spontaneous abortions and stillbirths (crude OR 9.9; 95% CI 3.3-29.4). CONCLUSION: Before and during an epidemic of acute B19 infection incidences were measured among pregnant women to be 1.0% and 13.5%, respectively. Three factors, significantly increasing the risk of acute B19, were identified as: having children at home; suffering from serious medical diseases; and having a stressful job. IgM positivity for parvovirus B19 was associated with negative outcome of pregnancy. PMID- 10826580 TI - Car seatbelts in pregnancy: the practice and knowledge of pregnant women remain causes for concern. AB - OBJECTIVE: To evaluate current knowledge and practice among pregnant women about the use of car restraint systems during pregnancy. DESIGN: A structured questionnaire. SETTING: District hospital antenatal clinic. PARTICIPANTS: Two hundred pregnant women attending for their routine mid-pregnancy anomaly scan. MAIN OUTCOME MEASURES: The women were asked about seatbelt and airbag usage, source of information about restraint systems and the legal requirements and recommendations regarding car safety systems. RESULTS: One hundred fifty-nine women (80%) completed the questionnaire. Ninety-eight percent wore seatbelts in the front, 68% wore seatbelts in the back of a car, 48% correctly identified where to place the seatbelt and 37% had received information on seatbelt use while pregnant. The latter were more likely to correctly position their restraints than those who had received no information (P = 0.03). Thirty-nine percent had an airbag system fitted and one woman had de-activated it. Fifty eight percent were aware seatbelt use was a legal requirement for driver and passengers when fitted. CONCLUSIONS: Many pregnant women are ignorant of the correct usage of seatbelts, their positioning and legal requirements. This puts their own wellbeing, and that of the fetus, at risk. Women should receive written instructions and advice from well informed health professionals and be encouraged always to wear a correctly positioned seatbelt. Education increases compliance and correct usage. PMID- 10826581 TI - Preterm delivery for maternal or fetal indications: maternal morbidity, neonatal outcome and late sequelae in infants. AB - OBJECTIVE: To assess maternal morbidity, and neonatal outcome and especially long term sequelae in infants born preterm due to maternal or fetal indications. DESIGN: Analysis of retrospective cohort. SETTING: Oulu University Central Hospital, Finland. POPULATION: One hundred and three women, who were between the 24th and the 33rd week of pregnancy, delivered by caesarean delivery because of maternal or fetal indications. They were matched with 103 women who had spontaneous preterm delivery at corresponding gestational weeks between 1990 1997. MAIN OUTCOME MEASURES: Maternal morbidity, reasons for caesarean delivery, neonatal mortality and morbidity rates, and later development of the infants. RESULTS: Pre-eclampsia was diagnosed in 57% of the women in the indicated group and only in one woman in the control group. All infants in the indicated group and almost a third in the control group were born by caesarean birth; the main indication was threatening fetal asphyxia. There was a significant difference in neonatal mortality rates between the groups (175 vs 78 per thousand live births in the indicated vs control infants; RR 2.3, 95% CI 1.02, 4.9) and the main cause of death was respiratory insufficiency: 64% in the indicated group and 22% in the controls; RR 2.9, 95% CI 0.8, 10. Respiratory distress syndrome occurred more often (73% vs 53%, RR 1.4, 95% CI 1.1, 1.7) and it was more severe and more complicated in infants in the indicated group, compared with those in the control group. Symptomatic chronic lung disease at one year of age was more common in infants in the indicated group than in the control group (15% vs 3%; RR 4.6, 95% CI 1.4, 15.9). CONCLUSIONS: Not only the risks of neonatal mortality and morbidity but also long term pulmonary consequences, appear to be greater in infants born preterm by indicated delivery than in preterm infants born spontaneously at corresponding weeks. PMID- 10826582 TI - Views of Finnish doctors on fetal screening. AB - OBJECTIVE: To investigate the views of Finnish doctors concerning fetal screening. DESIGN: Anonymous, questionnaire survey conducted in 1996-1997. POPULATION: A representative sample of gynaecologists, paediatricians and general practitioners in Finland. Both leading doctors and ordinary practitioners were included. RESULTS: Most doctors said that serum screening for Down's syndrome and ultrasound screening for structural abnormalities should be available for all pregnant women. In response to more direct questions, doctors acknowledged many drawbacks to Down's serum screening, notably the worry due to false positives. Only a few were against abortion, and a fifth said fetal screening is partly based on a eugenic ideology. There were some differences between the different doctor groups, but the overall impression was of similarity rather than divergence, both between the different specialist groups, and by the position of the doctor (leading vs ordinary). CONCLUSIONS: Finnish doctors support current fetal screening, but many acknowledged resulting ethical, psychological, and social problems. PMID- 10826583 TI - The effect of immersion on haemodynamic and fetal measures in uncomplicated pregnancies of nulliparous women. AB - OBJECTIVE: To evaluate the effect of immersion in water on maternal haemodynamic measures and fetal heart rate patterns in healthy nulliparous women. DESIGN: A randomised cross-over study. PARTICIPANTS: Twenty pregnant women were studied between 26-29 weeks and between 34-37 weeks of gestation. METHODS: The women were either immersed in water for 35 minutes or rested on a bed for a similar period of time. Maternal blood pressure, thoracic electrical bioimpedance, urine and blood samples, and fetal heart rate patterns were obtained before and after immersion or bedrest. RESULTS: Compared with bedrest and pre-immersion values, immersion resulted in a significant decrease in blood pressure, which returned to baseline values within 1.5 hour. Urine production increased for a short period after immersion, but 24 hour samples showed no difference compared with bedrest. Sodium, potassium and creatinine levels in urine remained constant. No significant changes were found for the other haemodynamic measures. There were no effects on fetal heart rate nor on its variation. CONCLUSION: In healthy pregnant women immersion for 35 minutes has a clear effect on blood pressure and urine production. However, these effects are short lasting. PMID- 10826584 TI - Fetal haemodynamic changes in fetuses during fetal development evaluated by arterial pressure pulse and blood flow velocity waveforms. AB - OBJECTIVE: To study the relation between diameter pulse (pressure pulse) and flow velocity waveforms in the fetal descending aorta in fetuses with intrauterine growth retardation and acceleration. DESIGN: Blood flow through a tubular system can be expressed by the ratio of blood pressure to vascular resistance. Doppler ultrasound and a phase locked loop echo tracking system coupled to a B-mode ultrasonic imager (central frequency 3.5 MHz) were used to assess downstream resistance and change in blood pressure, respectively. SETTING: Tertiary referral unit in a teaching hospital PARTICIPANTS: Serial study between 21 and 40 weeks of 22 women with normally grown fetuses at intervals of four weeks; 25 women with small for gestational age fetuses with uteroplacental insufficiency (high umbilical artery pulsatility index); and six women with large for gestational age fetuses. MAIN OUTCOME MEASURES: We measured the maximum systolic and minimum diastolic diameter of the fetal descending aorta (the pulse amplitude) and then calculated the pulsatile waveform time integral above the least diastolic diameter (pulsatile area) and total waveform time integral (perfusion area). RESULTS: Normal fetal growth was associated with an increase in systolic and diastolic diameters in the fetal descending aorta with advancing gestational age. Pulse amplitude, pulsatile and perfusion areas increased with gestational age. The increased pulse amplitude and increased pulsatile area in association with a decrease of the pulsatility index in the descending aorta during the second trimester suggested that pulse pressure and stroke volume were increased. In the group with intrauterine growth retardation, systolic and diastolic diameters of the descending aorta and perfusion area were within the normal range. Moreover, there was an increase in the diastolic diameter per unit fetal weight and a decrease in the pulsatile area. In the large for gestational age fetuses, there was an increase in the pulse amplitude and pulsatile area. CONCLUSION: These findings suggest that in growth restricted fetuses there is an increase in diastolic pressure and a reduction in stroke volume, while in large for gestational age fetuses there is an increase in the pulse pressure and stroke volume. It is possible that measurement of pressure pulse waveform in relation to Doppler velocity waveform may be used to infer changes of blood flow. PMID- 10826585 TI - Uteroplacental blood flow and placental vascular endothelial growth factor in normotensive and pre-eclamptic pregnancy. AB - OBJECTIVE: To determine whether placental vascular endothelial growth factor (VEGF) is increased in pre-eclampsia. DESIGN: Prospective cohort study. SETTING: Royal Prince Alfred Hospital, Sydney, Australia. SAMPLE: Eleven normotensive women and eight women with pre-eclampsia matched for age and gestation. METHODS: Uterine artery Doppler ultrasound flow velocity profiles were recorded in the third trimester and resistance index calculated as (Vs-Vd)/Vs (Vs = peak systolic flow velocity, Vd = end diastolic flow velocity). Placental tissue at delivery was examined for VEGF distribution with avidin-biotin-peroxidase immunohistochemistry. RESULTS: Uterine resistance index [median (range)] was significantly increased in pre-eclamptic women (normotensive: 0.42 (0.36-0.51); pre-eclampsia: 0.59 (0.40-0.75); P = 0.005). Notching of the uterine artery waveform, consistent with a high resistance circulation, was evident in early diastole in five women with pre-eclampsia but only one normotensive woman (P = 0.013). Placental VEGF was increased in women with pre-eclampsia in the decidual trophoblast (normotensive: 34% (4-59) cells stained for VEGF; pre-eclampsia: 58% (15-95); P = 0.033) and in the villous syncytiotrophoblast (normotensive: VEGF count 1.4 arbitrary units (1.1-2.1); pre-eclampsia: 1.8 arbitrary units (1.4 2.2); P = 0.041). Analysis indicated that uterine artery resistance index was directly correlated with placental VEGF staining, mean arterial pressure and birthweight. CONCLUSIONS: Abnormal uterine artery Doppler ultrasound flow velocity profiles in pre-eclampsia indicate increased uteroplacental resistance. The associated increase in placental VEGF may represent a compensatory mechanism attempting to restore blood flow towards normal. PMID- 10826586 TI - The effect of local anaesthetic spray on the pain associated with local anaesthetic injection, prior to biopsy or loop diathermy to the cervix in the outpatient colposcopy clinic. AB - We evaluated the effectiveness of local anaesthetic spray prior to injection of local anaesthetic before biopsy or large loop excision of the transformation zone of the cervix at colposcopy. This was a prospective, randomised, double-blind placebo-controlled study involving 51 women. Pain or discomfort was measured using a four-point categorical scale and a visual analogue score. Our results show that the use of local anaesthetic spray has no effect on the pain or discomfort experienced by patients having local anaesthetic injections to the cervix, and cannot be recommended. PMID- 10826587 TI - Serum CA125 at 11-14 weeks of gestation in women with morphologically normal ovaries. AB - In a number of pregnant women ovarian cysts are found incidentally during the routine first trimester scan. These cysts may pose diagnostic difficulties, and the measurement of serum CA125 levels can be used to aid management. In this study we measured maternal serum CA125 levels in 188 women with uncomplicated pregnancies between 11-14 weeks of gestation. All women had morphologically normal ovaries observed on ultrasound examination. The median serum CA125 levels were 23.4 U/mL (range 2.2-166.3 U/mL, 95% reference interval 5.28-70.15) and did not change significantly with gestation. We conclude that CA125 levels are increased at 11-14 weeks of gestation and cut off values which are used to assess the nature of ovarian cysts in nonpregnant women cannot be applied to pregnant women at this gestation. PMID- 10826588 TI - Neonatal alloimmune thrombocytopenia due to anti-HPA 1a antibodies; the level of maternal antibodies predicts the severity of thrombocytopenia in the newborn. AB - Eleven thousand one hundred pregnant women were genotyped for human platelet antigen HPA 1, and 198 HPA 1bb women were followed in the pregnancy with quantitative assay for anti-HPA la antibodies. Antibodies were detected in 24 women, and nine children were born with severe thrombocytopenia (< 50x10(9)/L). All mothers with high levels of antibodies were delivered of children with severe thrombocytopenia. None of the newborn infants had clinical signs of intra-cranial haemorrhage. The level of maternal anti-HPA 1a antibodies is predictive for fetal thrombocytopenia and may be used in decisions related to time and mode of delivery. PMID- 10826589 TI - Alveolar echinococcosis with bulky peritoneal spread--a rare but important diagnosis in gynaecological practice. PMID- 10826590 TI - Wrong turn--dead end. PMID- 10826591 TI - Life threatening myocardial ischaemia associated with the use of prostaglandin E1 to induce abortion. PMID- 10826592 TI - Administration of low-dose aspirin to mothers with small for gestational age fetuses and abnormal umbilical Doppler studies to increase birthweight: a randomised double-blind controlled trial. PMID- 10826593 TI - Activin A, inhibin A, inhibin B and parturition: changes of maternal and cord serum levels according to the mode of delivery. PMID- 10826594 TI - Could tight garments cause endometriosis? PMID- 10826595 TI - Could tight garments cause endometriosis? PMID- 10826596 TI - Switchable columnar metallomesogens AB - Chiral columnar liquid crystals have recently appeared as a promising new type of ferroelectric materials. To date, all the columnar liquid crystals that have been reported to show ferroelectric switching consist of organic compounds. However, metal-containing liquid crystals open this field to a significant number of new structures and offer the possibility of adding to the ferroelectric behavior other properties inherent to the presence of metals in the structure, such as magnetism, as well as the use of new methods of characterization (EPR, synchrotron radiation, etc.). The potential of columnar metal-containing liquid crystals as ferroelectric materials has been demonstrated even though only a few organic columnar ferroelectric liquid crystals have been described. As a first approach to this type of material, this concepts article describes the results obtained with chiral metal beta-diketonates that show ferroelectric switching in the columnar mesophase. It has been shown that these materials have a helical columnar arrangement in the mesomorphic state, and a chiral superstructure has been proposed from circular dichroism studies. This type of supramolecular structure plays a fundamental role in the ferroelectric properties of these compounds. The discussion is mainly focused on the strategy employed for the molecular design, and on the interpretation of the mesophase structure and the electrooptic effect. The use of a diverse range of techniques, both those commonly used in the field of liquid crystals and those that are more specific will be highlighted, and the principles of these specific techniques are summarized together with a justification of their applicability to this study. PMID- 10826597 TI - New polyfunctional magnesium reagents for organic synthesis AB - The iodine-magnesium exchange reaction allows the preparation of polyfunctional aryl, heteroaryl, or alkenyl magnesium reagents at low temperature. These reagents display the typical reactivity of Grignard compounds and undergo various copper-catalyzed reactions such as allylation or 1,4-addition. Using this halogen metal exchange reaction, it was possible to generate polyfunctional magnesium reagents on the solid phase. PMID- 10826598 TI - Enhancement of Benzylic basicity by a fluorine substituent at the para- position: a case of lone pair/lone pair repulsion AB - The introduction of a halogen atom at any aromatic position of toluene considerably accelerates the base-promoted deprotonation of the methyl group. p Fluorotoluene is the only exception; proton abstraction from its benzylic site occurs approximately at one tenth of the rate found with toluene (at -75 degrees C). Lone-pair repulsion appears to be at the origin of the decrease in acidity. Chloro- and bromotoluenes instantaneously exchange benzylic hydrogen against metal when treated with solution of lithium 2,2,6,6-tetramethylpiperidide (LITMP) in diethyl ether in the presence of potassium tert-butoxide and N,N,N',N",N" pentamethyldiethylenetriamine at -100 degrees C. Due to extensive side reactions ("aryne" formation as a consequence of concomitant deprotonation of aromatic sites adjacent to the halogen atom), products can be isolated only in moderate yield (10-35%), but they are regioisomerically pure. PMID- 10826599 TI - Synthesis, structural characterisation, and Monte Carlo simulation of the magnetic properties of the 3D-stacked honeycomb Cs(n) AB - Two new polymeric manganese-azido systems with formula Cs(n)-[[Mn(N3)3]n] (1) and [[N(C2H5)4]n][[Mn2-(N3)5(H2O)]n] (2) were synthesised and structurally characterised. Compound 1 crystallises in the P2(1)/n group and consists of a three-dimensional system with end-to-end and end-on azido bridges with the caesium atoms in the holes of the net. Magnetically, compound 1 is a rare case of a three-dimensional network with alternate ferro-antiferromagnetic interactions. Compound 2 crystallises in the P1 group and consists of double chains of manganese atoms bridged by end-on and, the exceptional, (mu-1,1,1)-azido bridges. Magnetically, compound 2 shows net ferromagnetic behaviour. Exact fit of the magnetic data was performed for the two compounds by means of Monte Carlo simulations based on the Metropolis algorithm on sets of 10 x 10 x 10 (1) and 1 x 1 x 320 (2) S = 5/2 classical spin centres. PMID- 10826600 TI - Ring-hydrogen participation in the keto--enol isomerization of the acetophenone radical cation AB - Molecular ions obtained from acetophenone have been observed to undergo proton transfer reactions in competition with unimolecular blackbody dissociation in a Fourier transform ion cyclotron resonance spectrometer provided with an in situ high temperature blackbody source. The ionizing energy dependence of these two processes and generation of the enol molecular ion by fragmentation of butyrophenone reveal that the keto ion undergoes blackbody dissociation exclusively while the enol ion promotes fast proton transfer reactions and undergoes very slow blackbody induced dissociation. Experiments with labeled acetophenone either on the methyl group or on the ring reveal that the enol ions can transfer both H+ and D+ suggesting that the mechanism responsible for the tautomerization process of these radical cations may involve scrambling of the methyl and ring hydrogens, or more than one mechanism. Theoretical calculations at the B3LYP level predict that the most favorable pathway for unimolecular isomerization of the keto ion involves initial migration of an ortho hydrogen to the carbonyl. The subsequent rearrangement to the enol form is calculated to require enough internal energy that would allow hydrogen walk around the benzene ring in agreement with the experimental results. The possibility that isomerization may also occur by a direct 1,3-hydrogen migration is also explored in terms of possible excited electronic states of the ion. PMID- 10826601 TI - Polybenzimidazole as a promising support for metal catalysis: morphology and molecular accessibility in the dry and swollen state AB - Polybenzimidazole (PBI) in beaded form (250-500 microm) has been characterized in the dry state by scanning electron microscopy (SEM), BET, and nitrogen porosimetry. In the swollen state, it has been characterized by inverse steric exclusion chromatography (ISEC) in tetrahydrofuran, toluene, and water, by ESR of TEMPONE (2,2,6,6-tetramethyl-4-oxo-1-oxypiperidine), and pulse field gradient spin echo (PGSE) NMR spectroscopy, toluene, in tetrahydrofuran, ethanol and water. The dry-state results are in good agreement with the ISEC results obtained in tetrahydrofuran, toluene, and water with regard to the 40-80 nm macroporosity. The swelling-dependent surface area and pore volume detected by ISEC in toluene and water reveal the amphiphilic nature of PBI. PMID- 10826602 TI - Isomerizations between nitrosyl halides X-N=O and isonitrosyl halides X-O-N: a matrix-spectroscopic study AB - Irradiation of nitrosyl bromide BrNO (4) with light of the wavelength lambda = 248 nm and nitrosyl chloride CINO (6) with lambda = 193 nm in an argon matrix at 10 K leads to the corresponding isomers isonitrosyl bromide BrON (5) and isonitrosyl chloride CION (7). Both new compounds 5 and 7 have been identified by comparison of the experimental and calculated (BLYP/6-311 + G*) IR spectra. Nitrosyl fluoride FNO (8) could not be transferred into isonitrosyl fluoride FON (9). The back reactions 5-->4 and 7-->6 can be initiated by UV (lambda > 310 nm), visible or IR light. The retransformation also occurs spontaneously in the matrix at 10 K under exclusion of any UV/Vis or IR radiation. Surprisingly, the reaction rates of these spontaneous back reactions are temperature independent between 8.5 and 25 K. The mechanism of these processes is discussed. PMID- 10826603 TI - 17O NMR spectroscopic characterization and the mechanism of formation of alkyl hydrotrioxides (ROOOH) and hydrogen trioxide (HOOOH) in the low-temperature ozonation of isopropyl alcohol and isopropyl methyl ether: water-assisted decomposition AB - Low-temperature ozonation of isopropyl alcohol (1a) and isopropyl methyl ether (1b) in [D6]acetone, methyl acetate, and tert-butyl methyl ether at -78 degrees C produced the corresponding hydrotrioxides, Me2C(OH)(OOOH) (2a) and Me2C(OMe)(OOOH) (2b), along with hydrogen trioxide (HOOOH). All the polyoxides investigated were characterized for the first time by 17O NMR spectroscopy of highly 17O-enriched species. The assignment was confirmed by GIAO/MP2/6-31++G* calculations of 17O NMR chemical shifts, which were in excellent agreement with the experimental values. Ab initio density functional (DFT) calculations at the B3LYP/ 6-31G*+ZPE level have clarified the transition structure (TS1, deltaE = 7.4 and 10.6 kcalmol(-1), relative to isolated reactants and the complex 1a ozone, respectively) for the ozonation of 1a: this, together with the formation of HOOOH and some other products, indicates the involvement of radical intermediates (R*, *OOOH) in the reaction. The activation parameters for the decomposition of the hydrotrioxides 2a and 2b (Ea, = 23.5+/-1.5 kcalmol(-1), logA = 16+/-1.8) were typical for a homolytic process in which cleavage of the ROOOH molecule occurs to yield a radical pair [RO* *OOH] and represents the lowest available energy pathway. Significantly the lower activation parameters for the decomposition of HOOOH (Ea = 16.5+/-2.2 kcalmol(-1), logA = 9.5+/-2.0) relative to those expected for the homolytic scission of the HO-OOH bond [bond dissociation energy (BDE) = 29.8 kcalmol(-1), CCSD(T)/6-311++G**] are in accord with the proposal that water behaves as a bifunctional catalyst and therefore participates in a "polar" (non-radical) decomposition process of this polyoxide. A relatively large acceleration of the decomposition of the hydrotrioxide 2a in [D6]acetone, accompanied by a significant lowering of the activation energies, was observed in the presence of a large excess of water. Thus intramolecular 1,3 proton transfer probably also involves the participation of water and is similar to the mechanism proposed for the decomposition of HOOOH. This hypothesis was further substantiated by the B3LYP/6-31++ G*+ZPE calculations for the participation of water in the decomposition of CH3OOOH, which revealed two stationary points on the potential energy surface corresponding to a CH3OOOH-HOH complex and a six-membered cyclic transition state TS2. The energy barriers were comparable with those calculated for HOOOH, that is, deltaE = 15.0 and 21.5 kcalmol(-1) relative to isolated reactants and the CH3OOOH-HOH complex, respectively. PMID- 10826604 TI - Micrometrically controlled surface modification of Teflon by redox catalysis: electrochemical coupling between Teflon and a gold band ultramicroelectrode AB - Carbon-fluorine bonds of Teflon (polytetrafluoroethylene, PTFE) can be reduced electrochemically with the purpose of modifying its adhesive and wetting surface properties by micrometrically controlled surface carbonization of the material. This can be performed adequately by redox catalysis provided that the redox mediator couple has a sufficiently negative reduction potential. The process is investigated kinetically with benzonitrile as the mediator and a gold-band ultramicroelectrode mounted adjacent to a PTFE block, though separated from it by an insulating micrometric mylar gap. For moderate fluxes of reduced mediator, the whole device behaves as a generator-collector double-band assembly with a constant current amplification factor. This is maintained over long periods of time, during which the carbonized PTFE zones extends over distances that are much wider than the slowly expanding cylindrical diffusion layer generated at the gold microband electrode. This establishes that the overall redox catalysis proceeds through electronic conduction in the n-doped carbonized material. Thus, carbonization progresses at the external edge of the freshly carbonized surface in a diffusion-like fashion (dependence on the square root of time), while the redox-mediator oxidized form is regenerated at the carbonized PTFE edge facing to the gold ultramicroelectrode, so that the overall rate of carbonization is controlled by solution diffusion only. For larger fluxes of mediator, the heterogeneous rate of reduction and doping of PTFE becomes limiting, and the situation is more complex. A conceptually simple model is developed which predicts and explains all the main dynamic features of the system under these circumstances and allows the determination of the heterogeneous rate constant of carbon-fluorine bonds at the interface between the carbonized zone and the fresh PTFE. This model can be further refined to account for the effect of ohmic drop inside the carbonized zone on the heterogeneous reduction rate constants and henceforth gives an extremely satisfactory quantitative agreement with the experimental data. PMID- 10826605 TI - Preparation of a new carboranyl lactoside for the treatment of cancer by boron neutron capture therapy: synthesis and toxicity of fluoro carboranyl glycosides for in vivo 19F-NMR spectroscopy. AB - The synthesis of new ortho-carboranyl lactosides 8, 17, 19 and glucosides 22 and 23 for the use in boron neutron capture therapy is reported. Carboranyl lactosides 17 and 19 as well as the glucosides 22 and 23 contain a fluorine atom to allow a noninvasive determination of these compounds in tumor cells by 19F-NMR spectroscopy. In cloning efficiency tests on human bronchial carcinoma cells the carboranyl lactosides 17 and 19 displayed almost no cytotoxicity. Thus, the considerably cytotoxic carboranyl alcohol 11 is detoxified when linked to a sugar moiety such as in carboranyl glucoside 22. PMID- 10826606 TI - Heck reactions of iodobenzene and methyl acrylate with conventional supported palladium catalysts in the presence of organic and/or inorganic bases without ligands AB - The vinylation of iodobenzene with methyl acrylate has been studied with several supported palladium catalysts in N-methylpyrrolidone in the presence of triethylamine and/or sodium carbonate. The reaction can be performed in air without any solubilizing or activating ligands. It was found that significant amounts of palladium leach out into the solvent and these dissolved Pd species essentially catalyze the reaction. It is interesting, however, that almost all the palladium species in the solution can redeposit onto the surface of the supports after the reaction has been completed (at 100% conversion of iodobenzene). Thus, the catalysts were recyclable without loss of activity. The use of both inorganic and organic bases is very effective in the promotion of the palladium redeposition as well as in the enhancement of the reaction rate. For Heck reactions with bromobenzene and chlorobenzene it was found that the use of triethylamine and sodium carbonate increases the selectivity of the Heck coupling product (benzene is also produced for these two substrates), but the mixed bases do not affect the overall rate of reaction as much. PMID- 10826607 TI - The radical cation of anti-tricyclooctadiene and its rearrangement products AB - The anti dimer of cyclobutadiene (anti-tricyclo[4.2.0.0(2.5)]octa-3,7-diene, TOD) is subjected to ionization by gamma-irradiation in Freon matrices, pulse radiolysis in hydrocarbon matrices, and photoinduced electron transfer in solution. The resulting species are probed by optical and ESR spectroscopy (solid phase) as well as by CIDNP spectroscopy (solution). Thereby it is found that ionization of anti-TOD invariably leads to spontaneous decay to two products, that is bicyclo[4.2.0]octa-2,4,7-triene (BOT) and 1,4-dihydropentalene (1,4-DHP), whose relative yield strongly depends on the conditions of the experiment. Exploration of the C8H8*+ potential energy surface by the B3LYP/6-31G* density functional method leads to a mechanistic hypothesis for the observed rearrangements which involves a bifurcation between a pathway leading to the simple valence isomer, BOT*+, and another one leading to an unprecedented other valence isomer, the anti form of the bicyclo[3.3.0]octa-2,6-diene-4,8-diyl radical cation (anti-BOD*+). The latter product undergoes a very facile H-shift to yield the radical cation of 1,3a-dihydropentalene (1,3a-DHP*+) which ultimately rearrranges by a further H-shift to the observed product, 1,4-DHP*+. PMID- 10826608 TI - The radical cation of syn-tricyclooctadiene and its rearrangement products AB - The syn dimer of cyclobutadiene (tricyclo[4.2.0.0(2.5)]octa-3,7-diene, TOD) is subjected to ionization under different conditions and the resulting species are probed by optical and ESR spectroscopy. By means of quantum chemical modelling of the potential energy surfaces and the optical spectra, it is possible to assign the different products that arise spontaneously after ionization or after subsequent warming or illumination of the samples. Based on these findings, we propose a mechanistic scheme which involves a partitioning of the incipient radical cation of TOD between two electronic states. These two states engage in (near) activation-less decay to the more stable valence isomers, cyclooctatetraene (COT*+) and a bis-cyclobutenylium radical cation BCB*+. The latter product undergoes further rearrangement, first to tetracyclo[4.2.0.0(2,4).0(3,5]oct-7-ene (TCO*+) and eventually to bicyclo[4.2.0]octa-2,4,7-triene (BOT*+) which can also be generated photochemically from BCB*+ or TCO*+. The surprising departure of syn-TOD*+ from the least-motion reaction path leading to BOT*+ can be traced to strong vibronic interactions (second-order Jahn-Teller effects) which prevail in both possible ground states of syn-TOD*+. Such effects seem to be more important in determining the intramolecular reactivity of radical cations than orbital or state symmetry rules. PMID- 10826609 TI - Stereoselective interactions of a specialized antibody with cholesterol and epicholesterol monolayers. AB - The stereoselective recognition by monoclonal antibodies of two-dimensional monolayers of cholesterol spread at the air-water interface is presented. Using immunofluorescence, we show that one antibody, raised and selected against crystals of cholesterol monohydrate, specifically recognizes monolayers of cholesterol, but not monolayers of epicholesterol--its epimeric form. This demonstrates that stereoselective recognition also applies to protein-surface interactions. PMID- 10826610 TI - Quinones as co-catalysts and models for the surface of active carbon in the phosphovanadomolybdate-catalyzed aerobic oxidation of benzylic and allylic alcohols: synthetic, kinetic, and mechanistic aspects AB - Quinones have been considered as reactive compounds present on the surface of active carbon. Thus, the co-catalytic use of quinones combined with the phosphovanadomolybdate polyoxometalate, PV2Mo10O40(5-), has been studied as an analogue of the known PV2Mo10O405-/C catalyst in oxidative dehydrogenation reactions. From the synthetic point of view both biphasic the quinone (org) Na5PV2Mo10O40- (aq) and monophasic quinone (org)- 4Q5PV2Mo10O40-(org) [4Q = (nC4H9)4-N+] systems are effective for the selective oxidation of benzylic and allylic alcohols to their corresponding aldehydes. Kinetic measurements carried out on the model oxidative dehydrogenation of 4-methylbenzyl alcohol in the presence of p-chloranil, 4Q5PV2Mo10O40, and molecular oxygen showed that the reaction was non-elementary, although the 4-methylbenzyl alcohol oxydehydrogenation was the rate-determining step. ESR measurements showed the presence of the semiquinone of p-chloranil, probably as a complex with the polyoxometalate. This proposed complex was shown to be a more potent oxidant than p-chloranil. Thus, for the oxidation of 4-methoxytoluene the semiquinone complex was active, whereas p-chloranil alone was inactive. Beyond the importance of understanding quinone-phosphovanadomolybdate polyoxometalate-catalyzed reactions, insight gained from the formation of semiquinone active species can be applied for heterogeneous and aerobic oxidative transformations catalyzed by PV2Mo10O405- with carbon matrices as active supports. PMID- 10826611 TI - A family of polynuclear cobalt and nickel complexes stabilised by 2-pyridonate and carboxylate ligands AB - The synthesis and structural characterisation of a series of cobalt and nickel cages are reported. Eight of these structures contain a [M10(mu3-OH)6(eta2, mu3 xhp),(eta2, mu2-O2CR)6]2+ core (where M = Co or Ni; xhp = 6-chloro- or 6-methyl-2 pyridonate: R = Me, Ph, CHMe2, CH2Cl, CHPh2 or CMe3), where the ten metal atoms describe a centred-tricapped-trigonal prism (ttp). The cage contains six hydroxide ligands around the central metal, and the exterior is coated with pyridonate and carboxylate ligands. For four of the cages additional metal centres are found attached to the upper and/or lower triangular faces of the trigonal prism, generating dodeca- and undecanuclear cages. Three further cages are reported that contain a metal core based on an incomplete centred tetraicosahedron. These cages involve trimethylacetate as a ligand in company with either 6-methyl-2-pyridonate or 6-chloro-2-pyridonate. Comparison of these latter structures with the trigonal prisms reveal that they can be described as a pentacapped-trigonal prism missing one edge. Magnetic studies of three of the nickel cages with trigonal prismatic cores show spin ground states of S = 8, 4 and 2 for Ni12, Ni11 and Ni10 cages, respectively. PMID- 10826612 TI - The gas-phase Meerwein reaction AB - A systematic investigation of a novel epoxide and thioepoxide ring expansion reaction promoted by gaseous acylium and thioacylium ions is reported. As ab initio calculations predict, and 18O-labeling and MS3 pentaquadrupole experiments demonstrate, the reaction proceeds by initial O(S)-acylation of the (thio)epoxides followed by rapid intramolecular nucleophilic attack that results in three-to-five-membered ring expansion, and forms cyclic 1,3-dioxolanylium, 1,3 oxathiolanylium, or 1,3-dithiolanylium ions. This gas-phase reaction is analogous to a condensed-phase reaction long since described by H. Meerwein (Chem. Ber. 1955, 67, 374), and is termed as "the gas-phase Meerwein reaction"; it occurs often to great extents or even exclusively, but in some cases, particularly for the most basic (thio)epoxides and the most acidic (thio)acylium ions, proton transfer (eventually hydride abstraction) competes efficiently, or even dominates. When (thio)epoxides react with (thio)-acylium ions, the reaction promotes O(S)-scrambling; when epoxides react with thioacylium ions and the adducts are dissociated, it promotes S/O replacement. An analogous four-to-six membered ring expansion also occurs predominantly in reactions of trimethylene oxide with acylium and thioacylium ions. PMID- 10826613 TI - Enantioselective Diels-Alder approach to C-3-oxygenated angucyclinones from (SS) 2-(p-tolysulfinyl)-1,4-naphthoquinone. AB - Chiral racemic vinylcyclohexenes 2, bearing oxygenated substituents and/or a methyl group at the C-5 position of the cyclohexene ring, were submitted to Diels Alder reactions with enantiomerically pure (SS)-(2-p-tolylsulfinyl)-1,4 naphthoquinone [(+)-1]. The domino cycloaddition/pyrolytic sulfoxide elimination process led to the formation of enantiomerically enriched angularly tetracyclic quinones anti-6 and syn-7, which were obtained from the kinetic resolution of the racemic diene. In all cases, (SS)-(2-p-tolylsulfinyl)-1,4-naphthoquinone reacted from the less hindered face of the more reactive s-cis conformation, to form products in good enantiomeric excesses. Steric effects and torsional interactions in the corresponding approaches account for the observed pi-facial diastereoselectivities at both partners. The usefulness of this methodology is illustrated with the four-step totally asymmetric synthesis of the C-3-oxygenated angucyclinone derivative (-)-8-deoxytetrangomycin 10 in 26% overall yield and with 50% enantiomeric purity. PMID- 10826614 TI - A copper(I) oxygenation precursor in the entatic state: two isomers of a copper(I) compound of a rigid tetradentate ligand AB - Oxygenation of [CuI(L1)(NC-CH3)]+ (L1 = dimethyl 2,4-bis(2-pyridinyl)-3,7 diazabicyclo-[3.3.1]-nonane-9-on-1,5-dicarboxylate) leads to a relatively stable mu-peroxo-dicopper(II) product. The stability of this type of oxygenation product has been shown before to be the result of the square pyramidal geometry of L1; preorganization by a dinucleating ligand has been shown to increase the stability of the mu-peroxo-dicopper(II) compound. The structural data presented here indicate that destabilization of the copper(I) precursor is another important factor. There are two isomers of [CuI(L1)(NCCH3)]+; one is yellow, and the other is red. X-ray crystallography indicates that one pyridinyl donor is not coordinated in the yellow compound and that the red compound is 5-coordinate. In the light of the X-ray structure of the metal-free ligand and that of the corresponding copper(II) compound, it emerges that the ligand cavity is well suited for copper(II), whereas the copper(I) compounds are highly strained. This is supported by 1H NMR spectra of the copper(I) species where a fast dynamic process leads to line broadening and by electrochemical data, which indicate that the copper(II) products are exceptionally stable. Also presented are structural (copper(II)), electrochemical, and spectroscopic data (1H NMR, copper(I)) of the derivative [Cu(L2)(X)]n+ with a methyl substituent at the alpha-carbon atom of the two coordinated pyridinyl groups (L2 = dimethyl 2,4-bis(2-pyridinyl-6-methyl) 3,7-diazabicyclo-[3.3.1]-nonane-9-on-1,5-dicarboxylate). There are two structural forms of [CuII(L2)(X)]n+ (X = NCCH3, Cl), which depend on the steric demand of the fifth donor X. For both, van der Waals repulsion leads to a destabilization of the copper(II) products, and this is also evident from an increase in the reduction potential (-110 mV vs. -477 mV, Ag/AgNO3). PMID- 10826615 TI - Rapid access to infrared reference spectra of arbitrary organic compounds: scope and limitations of an approach to the simulation of infrared spectra by neural networks AB - Substance identification by infrared spectroscopy is performed by comparison of the experimental spectrum with a reference spectrum from a printed compilation or a database. If the analyzed compound can not be found in a database the corresponding reference spectrum has to be simulated. In order to achieve this, several reasonable candidates of structures for the compound at hand have to be conceived and for all these, infrared spectra have to be developed. The simulated spectrum that is most similar to the experimental suggests the correct structure. A rapid spectrum prediction method based on neural networks has been developed that supplies reference spectra for any organic compound. The scope and limitations of this method will be discussed on a test set of 16 compounds representing a broad range of organic chemistry. PMID- 10826616 TI - Radial aplasia, poikiloderma and auto-immune enterocolitis--new syndrome or severe form of Rothmund-Thomson syndrome? AB - A syndrome is described in three isolated patients in whom the main features are bilateral radial aplasia, short stature, an inflammatory based 'elastic' pyloric stenosis, a pan-enteric inflammatory gut disorder that appears to be due to an autoimmune process, and poikiloderma. Other features in individual cases include cleft palate, micrognathia, anal atresia, patellar aplasia/hypoplasia and sensorineural deafness. This combination may represent a severe form of Rothmund Thomson syndrome or possibly a previously unrecognized condition. PMID- 10826617 TI - The KBG syndrome. AB - We report on two boys with a combination of short stature, a broad face, macrodontia and developmental delay. These features suggest that they have the KBG syndrome, a rare but distinctive phenotype. The pattern of inheritance remains uncertain and both autosomal dominant and X-linked recessive inheritance should be considered in genetic counselling. PMID- 10826618 TI - Congenital bowing of the long bones associated with camptodactyly, talipes equinovarus and agenesis of the corpus callosum. AB - We report a baby with congenital bowing of the long bones, camptodactyly, talipes equinovarus and radiological features resembling both Stuve-Wiedemann syndrome and Schwartz-Jampel syndrome type 2. The baby had, in addition, agenesis of the corpus callosum. This feature has not been reported in either of these syndromes. It is possible that this baby has a previously undescribed syndrome. PMID- 10826619 TI - Raine dysplasia: a Brazilian case with a mild radiological involvement. AB - We report a preterm male infant, the first child of a young consanguineous couple, whose physical examination revealed craniofacial disproportion with microcephaly, wide fontanelles, exophthalmos, low nasal root and hypoplastic nose, long philtrum, small mouth, high arched and narrow palate, micrognathia, dysplastic, low-set and rounded ears, short neck and, arthrogryposis. Postmortem findings included hypoplastic lungs. Radiological examinations showed mild and localized increased of bone density in the cranial vault and skull base and facial bones and undermodelled in the long bones. The above findings are characteristics of Raine dysplasia but the case reported here presents a mild bone involvement with only a localized bone sclerosis and absence of prenatal fractures. We discuss the possibility that this case represents an allelic mutation of the Raine gene. The consanguinity of the parents reinforces the hypothesis of autosomal recessive inheritance for this entity. PMID- 10826620 TI - Two patients with varying combinations of sternal cleft, haemangiomas, midline abdominal raphe, coarctation of the aorta with a right aortic arch. AB - We report two patients, one with sternal cleft, haemangiomas, supraumbilical midline raphe and the other with a sternal cleft, haemangiomas, coarctation of the aorta with a right aortic arch. PMID- 10826621 TI - Agenesis of the corpus callosum, camptodactyly and obesity. AB - We report a 16-year-old girl with agenesis of the corpus callosum, congenital heart disease (ventricular septal defect and abnormal venous return), 5th digit camptodactyly and obesity, born to first cousin Italian parents. Although this child shares features with acrocallosal syndrome and Camera-Marugo-Cohen syndrome, we think that this combination of anomalies allows the delineation of a new MCA/MR syndrome, possibly recessively inherited. PMID- 10826622 TI - Severe short stature, hyperphalangy of the index fingers, mental retardation and facial dysmorphism. AB - We present an adult female patient with a so far unreported syndrome of severe short stature, severe mental retardation, facial dysmorphism and hyperphalangy of the index fingers. Parental consanguinity suggests an autosomal recessive inheritance. PMID- 10826623 TI - Pseudotrisomy 13 syndrome in siblings. AB - We describe a brother and sister who both had holoprosencephaly, polydactyly, cardiac lesions and a normal karyotype. The parents were first cousins and a diagnosis of pseudotrisomy 13 syndrome is suggested. This report provides further support that the inheritance of pseudotrisomy 13 syndrome is autosomal recessive. PMID- 10826624 TI - Autosomal dominant ectrodactyly with sensorineural deafness. AB - Ectrodactyly or split hand/split foot malformation as it is now known, is a rare developmental disorder sometimes associated with other systemic malformations. Very few cases of congenital hearing loss coexisting with ectrodactyly have been reported world-wide. We profile one such family with the father and daughter being affected with bilateral foot ectrodactyly minor abnormalities of the 5th fingers and severe sensorineural hearing loss (SNHL). The inheritance in this family was likely to be autosomal dominant. There were no cytogenetically demonstrable structural aberrations in either case. PMID- 10826625 TI - Woodhouse and Sakati syndrome (MIM 241080): report of a new patient. AB - A 32-year-old male with Woodhouse Sakati syndrome (MIM 241080) is described. Two of the proband's brothers also have diabetes mellitus and similar facial features, however they are not dysarthric. An affected older brother died of an unknown cause at age 30. This confirms autosomal recessive inheritance. PMID- 10826626 TI - Juberg-Hayward syndrome: report of a case with cleft palate, distally placed thumbs and vertebral anomalies. AB - A 10-year-old male is described with Juberg-Hayward syndrome. He has a cleft palate with distally placed thumbs and multiple cervical hemivertebrae but does not have a cleft lip. The mode of inheritance is discussed. PMID- 10826627 TI - Ring chromosome 1 in a newborn. AB - A newborn girl with ring chromosome 1 is described. The letter reports the chromosome analysis of peripheral blood lymphocytes with 46,XX,r(1)(p36.3q44) and evidence of ring instability in umbilical cord fibroblasts. PMID- 10826628 TI - Duane retraction syndrome, a new feature in 22q11 deletion syndrome? AB - A female child with Duane retraction syndrome is described. A microdeletion on chromosome 22(q11) was discovered using FISH analysis. It is postulated that Duane retraction syndrome might be a new feature in 22q11 deletion syndrome. PMID- 10826629 TI - Oromandibular-limb hypogenesis spectrum and maternal lithium use. AB - A female is described with oromandibular-limb hypogenesis spectrum. The mother had taken lithium during the pregnancy. PMID- 10826630 TI - Post-axial limb defects with maternal sodium valproate exposure. AB - A male child is described with bilateral talipes equinovarus, absent fibulae with bowing and shortening of the tibiae and facial dysmorphic features. His mother was on a daily dose of 2000 mg sodium valproate for convulsions. This case raises the possibility that postaxial limb defects might be caused by intrauterine valproate exposure. PMID- 10826631 TI - VACTERL with hydrocephalus and isolated tracheo-oesophageal fistula in a first cousin. AB - A male infant is described of VACTERL with hydrocephalus where a female first cousin of this child had isolated oesophageal atresia with tracheo-oesophageal fistula. Possibly modes of inheritance are discussed. PMID- 10826632 TI - A fetus with VACTERL association, unilateral aplasia of the humerus and partial hemihypoplasia. AB - We report simultaneous occurrence of right partial hemihypoplasia and ipsilateral aplasia of humerus in a fetus with VACTERL association. Our case has further demonstrated the overlap between VACTERL and body hemihypoplasia. PMID- 10826633 TI - Unilateral proximal femoral focal deficiency and Hirschsprung disease. AB - A male with unilateral proximal femoral focal deficiency and Hirschsprung disease is described. PMID- 10826634 TI - The second unrelated case with isolated microcephaly and normal intelligence (microcephalia vera) AB - An isolated case of microcephaly with normal intelligence in a 16-year-old male is described and compared with previously reported cases. PMID- 10826635 TI - Peculiar facies, obesity, cleft lip and palate, growth hormone deficiency and mental retardation: a new syndrome? AB - A female child with peculiar facies, obesity, cleft lip and palate, growth hormone deficiency and mental retardation is described. The present case does not appear to fit any of the known syndromes. PMID- 10826636 TI - Growth deficiency, mental retardation and unusual facies. AB - An 8-year-old girl with growth deficiency, mental retardation, unusual facies and hypertrichosis is described. The case is compared with one described by Wiedemann et al. [(1989) Atlas of Clinical Syndromes, 2nd ed. London: Wolfe Publishing pp. 198-199]. PMID- 10826637 TI - Amperometric biosensors/sequential injection analysis system for simultaneous determination of S- and R-captopril. AB - Two amperometric biosensors based on L- and D-amino acid oxidase, respectively, are proposed for the simultaneous detection of S- and R-captopril in a sequential injection analysis system (SIA). The linear concentration ranges are: 0.4-1.6 micromol/l (S-captopril) and 120-950 nmol/l (R-captopril) with detection limits of 0.2 and 15 nmol/l, respectively. The biosensors/SIA system can be used reliably on-line in synthesis process control, for the simultaneous assay of S- and R-captopril with a frequency of 34 samples/h. PMID- 10826638 TI - Application of a luminescence-based pH optrode to monitoring of fermentation by Klebsiella pneumoniae. AB - The application of an optical sensor based on immobilization of the ruthenium complex [Ru(bpy)2(dhphen)]2+ (bpy = 2,2'-bipyridine, dhphen = 4,7-dihydroxy-1,10 phenanthroline) in Nafion to pH monitoring of fermentation by Klebsiella pneumoniae is described. Interference from the culture medium can be eliminated by addition of a black microporous filter membrane (pore size: 0.45 microm) on top of the sensing film. The response of this pH optrode was found to show good correlation with the conventional pH electrode. PMID- 10826639 TI - Highly sensitive optical chip immunoassays in human serum. AB - Over the past decade the ability of refractometric optical sensors to quantitatively measure a wide range of biomolecules has been demonstrated. These include proteins, nucleic acids, microorganisms, and in competitive formats small molecules such as drugs and pesticides. Furthermore, by using high refractive index nanoparticles to amplify the biomolecular binding signal, sensitivities approaching those of well established diagnostic assays have been achieved. However, to date it has not been possible to show rapid detection of analytes in complex bodily fluids such as serum, in a one-step procedure, due to the interference resulting from non-specific binding (NSB) to the sensor surface. We have carried out preliminary work on the control of interference due to NSB using an optical chip based on the Hartman interferometer. This interferometer configuration employs a reference sensing region that can be functionalized separately from the specific sensing region. Optical chips were stored dry after surface functionalization, and rehydrated in serum. The observed level of background drift in serum was reduced by an order of magnitude when an exposed reference was used, compared to a reference which was blind to the sample. An additional 70% reduction in signal drift in serum was achieved by controlling the surface chemistry of the optical chip using a biotin-poly(ethylene glycol) (PEG) blocking agent. This functionalization procedure was combined with a sandwich assay using gold nanoparticles to develop a one-step assay for human chorionic gonadotropin (hCG) in human serum with a detection limit of 0.1 ng/ml for a 35 min assay. PMID- 10826640 TI - A biosensor for the detection of gas toxicity using a recombinant bioluminescent bacterium. AB - A whole-cell biosensor was developed for the detection of gas toxicity using a recombinant bioluminescent Escherichia coli harboring a lac::luxCDABE fusion. Immobilization of the cells within LB agar has been done to maintain the activity of the microorganisms and to detect the toxicity of chemicals through the direct contact with gas. Benzene, known as a representative volatile organic compound, was chosen as a sample toxic gas to evaluate the performance of this biosensor based on the bioluminescent response. This biosensor showed a dose-dependent response, and was found to be reproducible. The immobilizing matrices of this biosensor were stored at 4 degrees C and were maintained for at least a month without any noticeable change in its activity. The optimal temperature for sensing was 37 degrees C. A small size of this sensor kit has been successfully fabricated, and found to be applicable as a disposable and portable biosensor to monitor the atmospheric environment of a workplace in which high concentrations of toxic gases could be discharged. PMID- 10826641 TI - Distribution and stability of membrane proteins in lipid membranes on solid supports. AB - Bacteriorhodopsin and the nicotinic acetylcholine receptor were biotinylated and reconstituted in lipidic membranes on silicon supports by fusion with proteoliposomes. The presence and distribution of the proteins were studied by binding with streptavidin. Radio-labelled streptavidin was employed for quantifying the amounts of protein remaining in the supported membranes after storage in buffer. The proteins within the membranes remained bound to the surface for weeks. The biological activity of reconstituted unlabelled receptor upon storage showed stability in membranes formed on silicon supports and a reduced stability when formed onto lipid monolayer covered supports. Atomic force microscopy studies on preparations in liquid showed bilayer structures but also attached, partly fused liposomes and membrane particles. In air, the surface was smoother and contained less of liposomes and more of stacked lipid layers. Preparations labelled with streptavidin conjugated to colloidal gold and imaged in air showed the proteins individually distributed, with no protein-rich patches or protein aggregates. PMID- 10826642 TI - Reagentless biosensors based on self-deposited redox polyelectrolyte oxidoreductases architectures. AB - Reagentless fructose and alcohol biosensors have been produced with a versatile enzyme immobilisation technique which mimics natural interactions and flexibility of living systems. The electrode architecture is built up on electrostatic interactions by the sequential adsorption of redox polyelectrolytes and redox enzymes giving rise to the efficient transformation of substrate fluxes into electrocatalytic currents. All investigated multilayer structures were self deposited on 3-mercapto-1-propanesulfonic acid monolayers self-assembled on gold electrodes. Fructose dehydrogenase, horseradish peroxidase (HRP) and the couple HRP-alcohol oxidase were electrochemically connected with a cationic poly[(vinylpyridine)Os(bpy)2Cl] redox polymer (RP) interface in a layer-by-layer self-deposited architecture. The dependence of the distance on the electrochemical response of this interface was also studied showing a clear decrease in the Faradaic current when the distance to the electrode surface was increased. The sensitivities obtained for each biosensor were 19.3, 58.1 and 10.6 mA M(-1) cm(-1) for fructose, H2O2 and methanol, respectively. The sensitivity values can be easily controlled by a rational deposition and manipulation of the charge in the catalytic layers. The electrostatic assembly of the electrochemical interface and the catalytic layers resulted in integrated biochemical systems in which mass transfer diffusion and heterogeneous catalytic and electron transfer steps are efficiently coupled and can be easily manipulated. PMID- 10826643 TI - Rise in background current over time in a subcutaneous glucose sensor in the rabbit: relevance to calibration and accuracy. AB - In order to calibrate a continuous glucose monitor, accurate determination of the background current (I0) is necessary, in part because I0 could change over time. We compared two methods of I0 measurement: (1), extrapolation of sensor output data (as a function of glucose level) to the intercept at zero glucose and (2) direct measurement of the output of a blank anode with no enzyme coat. We implanted telemetric sensors subcutaneously in rabbits and measured their outputs during tri-level glucose clamps once per week for 5 weeks. The two methods yielded similar results. I0 rose substantially over time and this increase reached significance during week 3 by the direct method but not until week 5 by the extrapolation method. Using the direct method, I0 rose from 3.41 (0.60-8.48 nanoamperes (nA), median and range) during week 1 to 13.42 (9.1-14.3) during week 5. Using the extrapolation method, I0 rose from 0.57 (0-16.7) during week 1 to 15.3 (12.2-21.6) during week 5. We conclude that I0 can rise over time. If this rise went undetected and was assumed to be stable, a one-point calibration procedure would overestimate glycemia in the hypoglycemic range, i.e. fail to appreciate the severity of hypoglycemia. It is recommended that during validation of a chronic glucose sensor, I0 be measured sequentially over time. PMID- 10826644 TI - Patterning multiple antibodies on polystyrene. AB - A method for patterning different antibodies on polystyrene is described. The polystyrene surface is coated with a material resistant to antibody adsorption and the coating material is selectively removed from the region where antibody adsorption is desired, exposing clean polystyrene. Upon immersion of the substrate in antibody solution, antibodies adsorb to the clean polystyrene, but not to the coated areas of the surface. Two methods have been used to remove the coating: ion beam sputtering and mechanical etching. The pattern was verified using gold-labeled antigen in conjunction with X-ray photoelectron spectroscopy and with FITC-labeled antigen and fluorescence microscopy. Substrates patterned in this way could be used in conjunction with a charge-coupled detector for a multi-analyte biosensor. This patterning technique is suitable for applications in which a fast, inexpensive fabrication process is necessary, for example, in single-use sensors. PMID- 10826645 TI - Sol-gel based glucose biosensors employing optical oxygen transducers, and a method for compensating for variable oxygen background. AB - Various types of thin-film glucose biosensors based on the use of the enzyme glucose oxidase (GOx) have been developed. The luminescent oxygen probe Ru(dpp)- whose emission is quenched by oxygen--is used to measure the consumption of oxygen. Three different combinations of oxygen transducer and sol-gel immobilized GOx were tested. In the first, GOx was sandwiched between a sol-gel layer doped with Ru(dpp) and a second sol-gel layer composed of pure sol-gel (the 'sandwich' configuration). In the second, a sol-gel layer doped with Ru(dpp) was covered with sol-gel entrapped GOx (the 'two-layer configuration'). In the third, both GOx and a sol-gel powder containing GOx were incorporated into a single sol-gel phase (the 'powder configuration'). In all cases, it was found to be essential to add sorbitol which results in a more porous sol-gel in which diffusion is not impaired. The sandwich configuration provides the highest enzyme activity and the largest dynamic range (0.1-15 mM), but suffers from a distinct decrease in sensitivity upon prolonged use. The two-layer configuration has the fastest response time (t90 = 50 s), while the 'powder configuration' provides the best operational lifetime. The storage stability of all configurations exceeds 4 months if stored at 4 degrees C. In an Appendix, equations are derived which describe the response of such sensors, how the effect of varying oxygen supply can be compensated for by making use of two sensors, one sensitive to oxygen only, the other to both oxygen and glucose, and how such sensors can be calibrated using two calibrators only. PMID- 10826646 TI - Development of highly selective and stable potentiometric sensors for formaldehyde determination. AB - Two types of biosensors selective to formaldehyde have been developed on the basis of pH-sensitive field effect transistor as a transducer. Highly or partially purified alcohol oxidase (AOX) and the permeabilised cells of methylotrophic yeast Hansenula polymorpha (as a source of AOX) have been used as sensitive elements. The response time in steady-state measurement mode is in the range of 10-60 s for the enzyme-based sensors and 60-120 s for the cell-based sensor. When measured in kinetic mode the response time of all biosensors developed was less than 5 s. The linear dynamic range of the sensor output signals corresponds to 5-200 mM formaldehyde for highly and partially purified alcohol oxidase, and 5-50 mM formaldehyde for the cells. The operational stability of the biosensors is not less than 7 h, and the relative standard deviation of intra-sensor response is approximately 2 and 5% for the enzyme- and cell-based sensors, respectively. When stored at 4 degrees C, the enzyme and cell sensor responses have been found stable for more than 60 and 30 days, respectively. Both types of biosensors demonstrate a high selectivity to formaldehyde with no potentiometric response to primary alcohols, including methanol, or glycerol and glucose. The possible reasons of such unexpected high selectivity of AOX-based FET-sensors to formaldehyde are discussed. The influence of the biomembrane composition and the effect of different buffers on the sensor response to formaldehyde are also discussed. PMID- 10826647 TI - Protein profiled features patterned via confocal microscopy. AB - Protein patterns were printed using conventional microlithographic materials in a bilayer arrangement and unconventional exposure tools. The bilayer resist stack consisted of a lower poly(tert-butyl methacrylate) layer and an upper diazonaphtoquinone/novolak layer. The protein features were printed in either 'contact printing', or 'step and repeat' mode. The latter printing mode can be managed in a flow-cell consisting of a standard microscope slide and cover slip, spaced apart by about 20 microm, as follows: (i) the exposure step is carried out in the cell using focused 488 nm beam of a confocal laser scanning microscope; (ii) the development step is performed by flowing the photoresist developer through the cell; (iii) the selective deposition of the protein (FITC-labelled avidin) is achieved via the flow of the protein solution through the cell until a desired contrast has been reached; (iv) the control of the process is assured using on-line monitoring of the photo-activated red fluorescence of the developing resist layer, and of the green fluorescence of the FITC-protein patterns, respectively. The protein printing technique uses equipment routinely available in biological laboratory. The 'step and repeat' patterning yields high and controllable resolution. The process can be applied in the fabrication of medical microanalysis devices. PMID- 10826648 TI - Hydrogen-ion binding of polycytidylic acid immobilized between Langmuir layers of dimethyldioctadecylammonium (DODA) in thin multilayer films. AB - The report describes the study of hydrogen-ion binding of Langmuir-Blodgett films contained with polycytidylic acid. A variety of multilayer films are analyzed and their UV absorption spectra are recorded. Poly (C) molecules established between dimethyldioctadecylammonium (DODA) layers are shown to exist in double stranded and semiprotonated form, independent of the pH value of the solution from which the films were made. A large hysteresis was found between forward and back proton titration of poly(C) immobilized in the LB films. This hysteresis points to a marked transference of both types of molecules during the film titration. This behavior also depends upon the types of molecules from which the films were made. PMID- 10826649 TI - Current awareness in biosensors & bioelectronics. PMID- 10826650 TI - Spontaneous hydrolysis of 4-trifluoromethylphenol to a quinone methide and subsequent protein alkylation. AB - 4-Trifluoromethylphenol (4-TFMP) was cytotoxic to precision-cut rat liver slices as indicated by loss of intracellular potassium. Intracellular glutathione levels decreased and fluoride ion levels increased in a time and concentration-dependent manner. The cytotoxicity of 4-TFMP did not appear to be due to the release of fluoride, however, since equimolar concentrations of sodium fluoride or potassium fluoride were not toxic. The ortho isomer (2-TFMP), which had a threefold slower rate of fluoride release, was much less toxic to liver slices. In incubations without slices, 4-TFMP spontaneously hydrolyzed in aqueous buffer at physiological pH to form 4-hydroxybenzoic acid via a quinone methide intermediate. The quinone methide was trapped by the addition of glutathione. Analysis of the glutathione adduct indicated that all of the fluorine atoms were lost during the hydrolysis, yielding a cresol derivative with the glutathione moiety attached to a benzylic carbonyl group. The glutathione conjugate was the primary product formed at low alkylphenol/glutathione ratios; however, at higher 4-TFMP concentrations additional unidentified products were observed. 4-TFMP also inhibited the in vitro enzyme activity of purlfied glyceraldehyde-3-phosphate dehydrogenase, a sulfhydryl-dependent enzyme, in a time and concentration dependentmanner. Loss of thiol residues closely paralleled the loss in enzyme activity. The coaddition of glutathione prevented 4-TFMP-induced loss of enzyme activity. The cytotoxicity of 4-TFMP therefore appears to be due to spontaneous quinone methide formation and subsequent alkylation of cellular macromolecules. PMID- 10826651 TI - Transcriptional blockages in a cell-free system by sequence-selective DNA alkylating agents. AB - There is considerable interest in DNA sequence-selective DNA-binding drugs as potential inhibitors of gene expression. Five compounds with distinctly different base pair specificities were compared in their effects on the formation and elongation of the transcription complex from the lac UV5 promoter in a cell-free system. All were tested at drug levels which killed 90% of cells in a clonogenic survival assay. Cisplatin, a selective alkylator at purine residues, inhibited transcription, decreasing the full-length transcript, and causing blockage at a number of GG or AG sequences, making it probable that intrastrand crosslinks are the blocking lesions. A cyclopropylindoline known to be an A-specific alkylator also inhibited transcription, with blocks at adenines. The aniline mustard chlorambucil, that targets primarily G but also A sequences, was also effective in blocking the formation of full-length transcripts. It produced transcription blocks either at, or one base prior to, AA or GG sequences, suggesting that intrastrand crosslinks could again be involved. The non-alkylating DNA minor groove binder Hoechst 33342 (a bisbenzimidazole) blocked formation of the full length transcript, but without creating specific blockage sites. A bisbenzimidazole-linked aniline mustard analogue was a more effective transcription inhibitor than either chlorambucil or Hoechst 33342, with different blockage sites occurring immediately as compared with 2 h after incubation. The blockages were either immediately prior to AA or GG residues, or four to five base pairs prior to such sites, a pattern not predicted from in vitro DNA-binding studies. Minor groove DNA-binding ligands are of particular interest as inhibitors of gene expression, since they have the potential ability to bind selectively to long sequences of DNA. The results suggest that the bisbenzimidazole-linked mustard does cause alkylation and transcription blockage at novel DNA sites. in addition to sites characteristic of untargeted mustards. PMID- 10826652 TI - Characterization of the major DNA adducts in the liver of rats chronically exposed to tamoxifen for 18 months. AB - Our previous study has shown that chronic exposure to tamoxifen (TAM) induced formation of high levels of DNA adducts in the liver, the target tissue of TAM induced carcinogenesis in rats. One of the major DNA adducts (spot 1), as detected by 32P-postlabeling, accounted for 53% of the total adducts. To characterize this major adduct, the current study has compared spot 1 with two previously identified TAM-DNA adducts, i.e. alpha-TAM-N2-deoxyguanine (alpha-TAM N2-dG) and alpha-N-desmethyl TAM-N2-deoxyguanine (alpha-N-dmTAM-N2-dG) by various rechromatography methods. It was found that spot 1 was further resolved into two fractions during rechromatography analysis, one fraction co-migrated with the alpha-TAM-N2-dG and the other fraction co-migrated with the alpha-N-dmTAM-N2-dG. These findings have demonstrated that chronic exposure to tamoxifen induced the same major DNA adducts, i.e. alpha-TAM-N2-dG and alpha-N-dmTAM-N2-dG as those detected in acutely exposed rats. PMID- 10826653 TI - Effect of the microsomal system on interconversions between hydroquinone, benzoquinone, oxygen activation, and lipid peroxidation. AB - Our previous results indicated that cytochrome P450 destruction by benzene metabolites was caused mainly by benzoquinone (Soucek et al., Biochem. Pharmacol. 47 (1994) 2233-2242). The aim of this study was to investigate the interconversions between hydroquinone, semiquinone, and benzoquinone with regard to both spontaneous and enzymatic processes in order to test the above hypothesis. We have also studied the participation of hydroquinone and benzoquinone in OH radicals formation and lipid peroxidation as well as the role of ascorbate and transition metals. In buffered aqueous solution, hydroquinone was slowly oxidized to benzoquinone via a semiquinone radical. This conversion was slowed down by the addition of NADPH and completely stopped by microsomes in the presence of NADPH. Benzoquinone was reduced to semiquinone radical at a significantly higher rate and this conversion was stimulated by NADPH and more effectively by microsomes plus NADPH while semiquinone radical was quenched there. In microsomes with NADPH. both hydroquinone and benzoquinone stimulated the formation of OH radicals but inhibited peroxidation of lipids. Ascorbate at 0.5-5 mM concentration also produced significant generation of OH radicals in microsomes. Neither hydroquinone nor benzoquinone did change this ascorbate effect. On the contrary, 0.1-1.0 mM ascorbate stimulated peroxidation of lipids in microsomes whereas presence of hydroquinone or benzoquinone completely inhibited this deleterious effect of ascorbate. Iron-Fe2+ apparently played an important role in lipid peroxidation as shown by EDTA inhibition, but it did not influence OH radical production. In contrast, Fe3+ did not influence lipid peroxidation, but stimulated OH radical production. Thus, our results indicate that iron influenced the above processes depending on its oxidation state, but it did not influence hydroquinone/benzoquinone redox processes including the formation of semiquinone. It can be concluded that interconversions between hydroquinone and benzoquinone are influenced by NADPH and more effectively by the complete microsomal system. Ascorbate, well-known antioxidant produces OH radicals and peroxidation of lipids. On the other hand, both hydroquinone and benzoquinone appear to be very efficient inhibitors of lipid peroxidation. PMID- 10826654 TI - Heterogenous effects of anthraquinones on drug-metabolizing enzymes in the liver and small intestine of rat. AB - The induction of a variety of drug-metabolizing enzymes by six anthraquinones (AQs) has been investigated in the liver and small intestine of rat. In the liver, the intragastric administration for 3 days of 100 mg/kg 9,10-anthraquinone (9,10-AQ). 1-hydroxy-AQ, 1,4-dihydroxy-AQ, but not 1,2-dihydroxy-AQ and 2-carboxy AQ, resulted in a significant induction of the UDP-GT, DT-diaphorase, P450 1A linked monooxygenase activities and in particular the methoxyresorufin-O demethylase (MEROD), an activity dependent on P450 1A2. Immunoblot analysis indicated that 1-hydroxy-AQ and 1,4-dihydroxy-AQ induced P450 1A2 but not 1A1 and 9,10-AQ induced both P4501A2 and P4502B. Northern blotanalysis, using a cDNA probe for CYP 1A1 and CYP 1A2, confirmed that the AQs induce CYP 1A2 but not 1A1 mRNA. In the mucosa of small intestine, none of the above-mentioned enzymatic activities were enhanced following AQ administration. The induction mechanism of the hepatic enzymes by AQs is not known and it deserves a further study as it might be independent from the activation of the Ah-receptor as reported for other tricyclic compounds. The results from inhibition experiments showed that the hydroxylated AQs were strong inhibitors of P450 1A2-dependent monooxygenases. This suggests that long-term ingestion of certain AQs, may affect the toxicity of other components present in the diet through the hepatic induction or inhibition of P450 1A2. PMID- 10826655 TI - Overview of the mechanism of action of lithium in the brain: fifty-year update. AB - Since its discovery, lithium has been shown to act upon various neurotransmitter systems at multiple levels of signaling in the brain. Lithium, affecting each neurotransmitter system within complex interactive neuronal networks, is suggested to restore the balance among aberrant signaling pathways in critical regions of the brain. Recent molecular studies have revealed the action of lithium on signal transduction mechanisms, such as phosphoinositide hydrolysis, adenylyl cyclase, G protein, glycogen synthase kinase-3beta, protein kinase C, and its substrate myristoylated alanine-rich C kinase substrate. Such effects are thought to trigger long-term changes in neuronal signaling patterns that account for the prophylactic properties of lithium in the treatment of bipolar disorder. Through its effects on glycogen synthase kinase-3beta and protein kinase C, lithium may alter the level of phosphorylation of cytoskeletal proteins, which leads to neuroplastic changes associated with mood stabilization. Chronic lithium regulates transcriptional factors, which in turn may modulate the expression of a variety of genes that compensate for aberrant signaling associated with the pathophysiology of bipolar disorder. Future studies on long-term neuroplastic changes caused by lithium in the brain will set the stage for new drug-discovery opportunities. PMID- 10826656 TI - The psychopharmacologic specificity of the lithium ion: origins and trajectory. AB - This article examines the development of lithium therapy since its dramatic introduction into psychiatry in 1949. Since that time, lithium has been examined in the treatment of a variety of neuropsychiatric conditions, but it is in the treatment of bipolar disorder that it is most effective. This suggests that it has specificity in the treatment of this disorder. These findings are very relevant as they suggest that understanding the mechanism of action of lithium in bipolar disorder may hold keys to elucidating its pathophysiology and to developing newer and more effective treatments. We review the published data on the effectiveness of lithium in bipolar disorder and various neuropsychiatric conditions and also the available data on anticonvulsants and newer therapeutic agents in the treatment of this condition. PMID- 10826657 TI - The longitudinal course of bipolar disorder. AB - Course of illness is central to our focus on bipolar disorder due to the lifelong nature of this illness in the majority of patients. In this overview, we highlight areas of consensus and debate on factors that impact course of illness. Findings on age at onset, psychiatric comorbidity, frequency of episodes, cycle pattern, rapid cycling, mixed symptoms, and precipitants of episodes including use of substances and antidepressants and lithium discontinuation are discussed. The diversity and range of presentation and even course of illness become quickly apparent in this review. Highlighting these factors rather than seeking a unifying theory should be a productive way to refine our ability to identify additive factors contributing to course of illness for patients with bipolar disorder. PMID- 10826658 TI - Bipolar disorder and attention-deficit/hyperactivity disorder in children and adolescents. AB - The relationship between bipolar disorder and attention-deficit/hyperactivity disorder (ADHD) in children and adolescents has been one of the most hotly debated topics in recent child psychiatry literature. At the heart of the matter is whether large numbers of children with bipolar disorder are being unrecognized or misdiagnosed. The differential diagnoses of juvenile-onset bipolar disorder can be complicated by many factors, but the most common clinical dilemmas seem to arise from overlapping symptomatology with ADHD and the differing treatment strategies these diagnoses imply. This article discusses the similarities and differences between these disorders with respect to phenomenology, epidemiology, family history, brain imaging, and treatment response. PMID- 10826659 TI - Efficacy of lithium in mania and maintenance therapy of bipolar disorder. AB - Lithium was introduced in 1949 as a treatment for mania, for which there is still the strongest evidence of its efficacy. It has consistently yielded better results in the treatment of mania than neuroleptics and carbamazepine and equivalent results to divalproex. Its efficacy in bipolar depression remains inadequately studied. Lithium also provides benefit in prophylaxis. However, the percentage of patients persistently benefited is low, because it has both low efficacy in many symptomatic and illness course presentations of the disorder and low tolerability. Converging evidence from clinical and animal studies indicates that a principal behavioral effect of lithium is reduction of motor activity. Lithium is increasingly used in combined treatment regimens, often thereby allowing lower, better tolerated dosing and complementary benefits from drugs with different profiles of action. PMID- 10826660 TI - In vivo imaging of the pharmacodynamics and pharmacokinetics of lithium. AB - The therapeutic efficacy of lithium for the long-term management of bipolar disorder is well recognized, along with the risk of lithium-induced toxicity. The author describes the current findings of in vivo functional neuroimaging techniques with respect to the pharmacokinetics and pharmacodynamics of lithium and their future potential to elucidate the drug distribution and neural mechanisms that produce its therapeutic effects. Brain Li nuclear magnetic resonance spectroscopy findings have disassociated postdose brain and blood lithium concentrations and suggest a pharmacokinetic basis for lithium response and nonresponse. The application of in vivo synaptic activity and neurochemical imaging is providing new knowledge related to the distributed neural activity associated with lithium response and is contributing to the critical human testing of neuroprotective and signal transduction models of lithium's therapeutic effects. PMID- 10826661 TI - Suicide and bipolar disorder. AB - Suicide, which is both a stereotypic yet highly individualized act, is a common endpoint for many patients with severe psychiatric illness. The mood disorders (depression and bipolar manic-depression) are by far the most common psychiatric conditions associated with suicide. At least 25% to 50% of patients with bipolar disorder also attempt suicide at least once. With the exception of lithium--which is the most demonstrably effective treatment against suicide-remarkably little is known about specific contributions of mood-altering treatments to minimizing mortality rates in persons with major mood disorders in general and bipolar depression in particular. Suicide is usually a manifestation of severe psychiatric distress that is often associated with a diagnosable and treatable form of depression or other mental illness. In a clinical setting, an assessment of suicidal risk must precede any attempt to treat psychiatric illness. PMID- 10826662 TI - Lithium in unipolar depression and the prevention of suicide. AB - Unipolar depression is a severe recurrent illness with high lifetime morbidity and premature mortality due to suicide. Numerous double-blind, placebo-controlled trials have shown that lithium is very effective at reducing relapses when given as maintenance therapy. It is also very effective when given as maintenance therapy after electroconvulsive therapy. It can be given once a day at night, and controlled trials have shown a 12-hour plasma lithium level between 0.5 and 0.7 mmol/L the most effective, with very slight side effects. Long-term studies of lithium maintenance therapy show a suicide rate of 1.3 suicides per 1000 patient years. This is much lower than comparative studies in long-term follow-up of untreated depression, which show about 5.5 suicides per 1000 patient years. Although it is neither feasible nor ethical to carry out double-blind studies on suicide reduction, the massive evidence showing a reduction in morbidity on lithium treatment suggests that systematic long-term lithium treatment of unipolar depression could considerably lower the suicide rate. PMID- 10826663 TI - The treatment of bipolar depression. AB - BACKGROUND: The treatment of bipolar depression represents a relatively understudied area in clinical psychiatry. The depressive phases of bipolar disorder can be very disabling, with significant associated comorbidity and suicide risk, impairment in functioning, and infringement on quality of life. We review the current evidence for the management of bipolar depression. METHOD: References for this review were obtained through MEDLINE searches of the medical literature on subjects pertaining to the treatment of bipolar depression. Search terms included bipolar depression, antidepressants, and bipolar disorder. Only publications in English are reviewed here. RESULTS: Lithium is currently the gold standard and most appropriate initial treatment for the depressive phase of bipolar disorder. Other mood stabilizers have demonstrated preliminary efficacy. Of the antidepressants, bupropion and the selective serotonin reuptake inhibitors may be associated with less risk of inducing hypomania, mania, and rapid cycling compared with tricyclic antidepressants. Monoamine oxidase inhibitors should be considered for patients with anergic bipolar depression. Electroconvulsive therapy has been shown to be highly efficacious. Other treatment modalities, including psychotherapy, sleep deprivation, phototherapy, and newer medications, require further research. CONCLUSIONS: Although the treatment of bipolar depression can be a complicated clinical task, the treatment armamentarium is expanding. Further research, especially in the form of randomized controlled trials, is warranted. Clinicians should be familiar with general guidelines for the use of psychopharmacologic agents for treating bipolar depression. PMID- 10826664 TI - Integration of pharmacotherapy and psychotherapy for bipolar disorder. AB - There is no question that pharmacotherapy is the treatment of choice for bipolar disorder. However. an integration of psychotherapeutic techniques with pharmacotherapy has been recommended by the American Psychiatric Association practice guideline for the treatment of bipolar disorder. Psychotherapy aims to address risk factors and associated features that are difficult to address with pharmacotherapy alone. The most common psychotherapeutic approaches added to pharmacotherapy for bipolar disorder include psychoeducation, individual cognitive-behavioral therapy, marital and family interventions, individual interpersonal therapy, and adjunctive therapies such as those for substance use. Each of these approaches is described in detail, and research regarding their efficacy is presented. PMID- 10826665 TI - Optimizing lithium treatment. AB - The purpose of this article is to discuss treatment of side effects that can occur during lithium therapy. Side effects from lithium are common but generally benign. For this article, I have divided the side effects into those that occur early, those that are late appearing, side effects related to drug interactions, and lithium toxicity. Side effects can decrease compliance. Lithium is a very effective drug for the stabilization of mood disorder in bipolar patients. Since side effects can affect compliance, recognition and treatment of early and late appearing side effects are important aspects of lithium pharmacotherapy. PMID- 10826666 TI - Lithium up-regulates the cytoprotective protein Bcl-2 in the CNS in vivo: a role for neurotrophic and neuroprotective effects in manic depressive illness. AB - Although mood disorders have traditionally been conceptualized as "neurochemical disorders," considerable literature from a variety of sources demonstrates significant reductions in regional central nervous system (CNS) volume and cell numbers (both neurons and glia) in persons with mood disorders. It is noteworthy that recent advances in cellular and molecular biology have resulted in the identification of 2 novel, hitherto completely unexpected targets of lithium's actions, discoveries that may have a major impact on the future use of this unique cation in biology and medicine. Chronic lithium treatment has been demonstrated to markedly increase the levels of the major neuroprotective protein bc1-2 in rat frontal cortex, hippocampus, and striatum. Similar lithium-induced increases in bc1-2 are also observed in cells of human neuronal origin and are observed in rat frontal cortex at lithium levels as low as approximately 0.3 mM. Bc1-2 is widely regarded as a major neuroprotective protein, and genetic strategies that increase bc1-2 levels have demonstrated not only robust protection of neurons against diverse insults, but have also demonstrated an increase in the regeneration of mammalian CNS axons. Lithium has also been demonstrated to inhibit glycogen synthase kinase 3beta (GSK-3beta), an enzyme known to regulate the levels of phosphorylated tau and beta-catenin (both of which may play a role in the neurodegeneration observed in certain forms of Alzheimer's disease). Consistent with the increases in bc1-2 levels and inhibition of GSK-3beta, lithium has been demonstrated to exert robust protective effects against diverse insults both in vitro and in vivo. These findings suggest that lithium may exert some of its long-term beneficial effects in the treatment of mood disorders via underappreciated neurotrophic and neuroprotective effects. To date, lithium remains the only medication demonstrated to markedly increase bc1-2 levels in several brain areas; in the absence of other adequate treatments, an investigation of the potential efficacy of lithium in the long-term treatment of several neurodegenerative disorders is warranted. Additionally, we suggest that a reconceptualization of the use of lithium in mood disorders may be warranted-namely, that the use of lithium as a neurotrophic/neuroprotective agent should be considered in the long-term treatment of mood disorders, irrespective of the "primary" treatment modality being used for the condition. PMID- 10826667 TI - Reduced suicide risk during lithium maintenance treatment. AB - BACKGROUND: About 20% of deaths of bipolar disorder patients are suicides associated with depressive or mixed episodes. Long-term lithium treatment may be associated with reduction of suicidal risk. METHOD: We reviewed studies and our previously reported data to quantify relationships of presence versus absence of lithium maintenance and suicides or suicide attempts in bipolar disorder patients. RESULTS: Results from 22 studies (1974-1998) yielded 7-fold lower suicidal rates for patients during long-term lithium treatment than for these patients when they were not receiving such treatment, patients lacking such treatment, or for patients after lithium discontinuation. CONCLUSIONS: Protection against suicide with lithium is incomplete, but rates of suicides plus attempts during lithium treatment may approach general population base rates. Better protection against bipolar depression is essential for limiting suicidal risk: alternatives to lithium require further study for effects on suicidal behavior. PMID- 10826668 TI - Detection in capillary electrophoresis. AB - A review of the four major, on-line, capillary electrophoresis (CE) detection modalities is presented. It is shown that each detection method, fluorescence, absorbance (conventional and nonconventional), electrochemical and refractive index, have distinct advantages and limitations when applied to analysis in a CE format. Various aspects of CE detection are considered and a perspective regarding the applicability of the technique is provided. It is shown that because of widely varying detection limits (ranging from single molecule to 10( 5) M) and detection scheme complexity, the particular application should dictate the selection of detection methodology in CE. PMID- 10826669 TI - Fluorescence line-narrowing detection in chromatography and electrophoresis. AB - A review of the basic aspects of fluorescence line-narrowing spectroscopy (FLNS) and its coupling with thin-layer chromatography (TLC) and polyacrylamide gel electrophoresis (PAGE) for off-line high-resolution low temperature spectral characterization is discussed. This is followed by a description of the on-line interfacing of capillary electrophoresis (CE) and capillary electrochromatography (CEC) with FLN detection. CE/ CEC-FLNS instrumentation and its applications for spectral identification of closely related analytes are also presented. Future prospects of micro and capillary high performance liquid chromatography (HPLC) with on-line high-resolution low temperature spectroscopic identification are considered. PMID- 10826670 TI - Near-infrared laser-induced fluorescence detection in capillary electrophoresis. AB - As capillary electrophoresis continues to focus on miniaturization, either through reducing column dimensions or situating entire electrophoresis systems on planar chips, advances in detection become necessary to meet the challenges posed by these electrophoresis platforms. The challenges result from the fact that miniaturization requires smaller load volumes, demanding highly sensitive detection. In addition, many times multiple targets must be analyzed simultaneously (multiplexed applications), further complicating detection. Near infrared (NIR) fluorescence offers an attractive alternative to visible fluorescence for critical applications in capillary electrophoresis due to the impressive limits of detection that can be generated, in part resulting from the low background levels that are observed in the NIR. Advances in instrumentation and fluorogenic labels appropriate for NIR monitoring have led to a growing number of examples of the use of NIR fluorescence in capillary electrophoresis. In this review, we will cover instrumental components used to construct ultrasensitive NIR fluorescence detectors, including light sources and photon transducers. In addition, we will discuss various types of labeling dyes appropriate for NIR fluorescence and finally, we will present several applications that have used NIR fluorescence in capillary electrophoresis, especially for DNA sequencing and fragment analysis. PMID- 10826671 TI - Applications of capillary electrophoresis with electrochemical detection in pharmaceutical and biomedical analyses. AB - As a high efficiency separation technique, capillary electrophoresis has been widely used in various fields of analytical science. This review discusses the applications of electrochemical detection systems combined with capillary electrophoresis in pharmaceutical and biomedical analysis. These detection methods mainly involve amperometric detection but also include conductivity detection and potentiometric detection. Its applications in the field are divided into six parts, including catechol compounds, thiols, amino acids and peptides, carbohydrates, general pharmaceuticals, and other related compounds. A relatively detailed discussion is described for each compound under the current studied. On this basis, we have suggested several conceivable directions for capillary electrophoresis with electrochemical detection in the future. PMID- 10826672 TI - A multireflection cell for enhanced absorbance detection in microchip-based capillary electrophoresis devices. AB - The design, fabrication and testing of a photolithographically fabricated, glass based multireflection absorbance cell for microfluidic devices, in particular microchip-based capillary electrophoresis (CE) systems is described. A multireflection cell was fabricated lithographically using a three-mask process to pattern aluminum mirrors above and below a flow channel in a chip, with 30 microm diameter optical entrance or exit apertures (one in each mirror) positioned 200 microm apart. Source and detector were positioned on opposite sides, and the metal mirrors were made 1 cm square, to reduce stray light effects. Calibration curves using bromothymol blue (BTB) with a 633 nm source (He:Ne laser) were linear to at least 0.5 absorbance units, with typical r2 values of 0.9997, relative standard deviations in the slopes of +/- 1.3%, and intercepts of zero within experimental error. Effective optical pathlengths of 50 272 microm were achieved, compared to single-pass pathlengths of 10-30 microm, corresponding to sensitivity enhancements (i.e., optical path length increase) of 5 to 10-fold over single-pass devices. Baseline absorbance noise varied within a factor of two in almost all devices, depending only weakly on path length. This device can give much higher absorbance sensitivity, and should be much easier to manufacture than planar, glass-based devices previously reported. PMID- 10826673 TI - Improving signal to background ratio for on-the-fly fluorescence lifetime detection in capillary electrophoresis. AB - On-the-fly fluorescence lifetime detection (OFLD) in capillary electrophoresis (CE) was previously demonstrated using a commercial multiharmonic Fourier transform (MHF) spectrofluorometer interfaced to a commercial CE system. This paper discusses optimization of the interface design for minimization of background fluorescence and scattered light, thereby maximizing the signal-to background ratio (S/B) of the dynamic measurement. Strategies included using various combinations of optical filters including a holographic filter and longpass or bandpass filters, tilting the capillary relative to the incident laser beam, employing a confocal design and adding an iris to remove out-of-focus light, using a microscope objective in the emission beam to increase the collection efficiency, and using square instead of ciruclar capillary columns. Significant improvements in S/B for on-column, on-the-fly detection of fluorescein in CE were achieved with most modifications. PMID- 10826674 TI - Quenched phosphorescence, a new detection method in capillary electrophoresis. AB - The applicability of quenched phosphorescence as a detection mode in capillary electrophoresis (CE) was explored for a number of analyte classes and buffer systems. The detection method is based on the quenching of biacetyl phosphorescence (biacetyl is a constituent of the CE buffer) by the analytes via various mechanisms (energy transfer, electron transfer and, possibly, hydrogen donation) and gives rise to negative peaks in the electropherograms. A number of buffers in the pH range 4.7-11.5, frequently used in CE, were tested for their compatibility with this detection mode. Borate, succinate, malonate, acetate, and phosphate buffers (pH 4.7-8.5) could be used without any problems. With a pH of ca. 8.5 or higher the baseline declined with time, while at a pH higher than 9.5 no signal at all was obtained. Obviously, the noise on the phosphorescence signal (i.e., the baseline) determines the ultimate analyte detection limits (LODs). The baseline signal-to-noise ratio, usually denoted as the dynamic reserve (DR), was enhanced ca. 25-fold compared to direct biacetyl excitation by sensitization of the biacetyl phosphorescence by 1,5-naphthalenedisulfonic acid, and by application of a total emission mirror (TEM). A concentration of 1 x 10(-3) M 1,5 naphthalenedisulfonic acid was found to be optimal. For the buffer systems considered, the DR was typically ca. 300-600 under optimized conditions (noise defined as 1 x sigma). Investigated analytes include naphthalenesulfonic acids (NS), nitrophenols, hydroxybenzoic acids, amino acids, and dithiocarbamates (DTCs.). For most of these, the LODs were in the 10(-7)-10(-8) M range, which is significantly lower than with direct or indirect absorption detection. PMID- 10826675 TI - Indirect photomeric detection of anions in capillary electrophoresis using dyes as probes and electrolytes buffered with an isoelectric ampholyte. AB - The use of highly absorbing anionic dyes as probes and isoelectric ampholytes as buffers in background electrolytes (BGEs) combined with the use of a light emitting diode (LED) as a light source has been studied for ultrasensitive indirect photometric detection in capillary electrophoresis (CE). Potential dyes and buffers were evaluated based on characteristics relevant to indirect photometric detection principles, such as the electrophoretic mobility of the probe dye, its solubility and adsorption behaviour, and the isoelectric point and buffering capacity of the ampholytic buffer. Two dyes, tartrazine and naphthol yellow S, and histidine as the ampholytic buffer, were selected for detailed investigation. Purification of the probes was vital to avoid anionic impurities interfering with the detection. For the electrolytes containing a purified probe (0.5 mM) and histidine as the isoelectric buffer (p/ 7.7), hydroxypropylmethyl cellulose (approximately 0.05%) was effective in suppression of the electroosmotic flow (EOF). Analytical method performance characteristics were determined. For both probes, experimentally determined mobilities were generally close to literature values, excellent peak shapes and separation efficiencies of up to 298 000 theoretical plates were obtained, and detection limits were generally at the sub-microM level. For the naphthol yellow S-histidine BGE, linearity and reproducibility were also evaluated, with excellent linearity being observed over a range of 5-500 microM, and reproducibility (relative standard deviation, RSD) less than 1% for migration times and 2-8% for normalised peak areas. The approach developed was applied successfully to several real samples including tap water, mineral waters, and beer. PMID- 10826676 TI - Electrofilament deposition and off-column detection of analytes separated by capillary electrophoresis. AB - Capillary electrophoresis interfaced with electrospray is a convenient technique for continuously transferring column effluent from capillary-to-planar format. Conditions are optimized to produce a narrow (approximately 20 microm) liquid filament (electrofilament), which is capable of depositing spatially focused bands with track widths that are routinely 100 microm. A fiber optic-based, laser induced fluorescence cell is employed to monitor the separation on-column while the separated bands are deposited onto a moving substrate. The photodetection of deposited bands is accomplished by using either a charge-coupled device camera or a photomultiplier tube. Deterioration of on-column separation performance is observed when the electrofilament voltage is applied. Elevating the inlet of the capillary column, to provide hydrodynamic flow, restores separation performance. Substrate temperature and translational rates are optimized with respect to both off-column separation efficiency and signal intensity. Off-column separation efficiencies of 65 000 plates per meter were achieved. A linear dynamic range of 10(3) and a limit of detection of 10(-8) M were obtained for kiton red deposited onto a reversed phase thin-layer chromatography plate. To demonstrate the applicability of this technique to more complex separation solutions, a dye mixture was successfully separated and deposited with sodium dodecyl sulfate in the running buffer. PMID- 10826677 TI - Construction and evaluation of a capillary array DNA sequencer based on a micromachined sheath-flow cuvette. AB - A capillary array electrophoresis DNA sequencer is reported based on a micromachined sheath-flow cuvette as the detection chamber. This cuvette is equipped with a set of micromachined features that hold the capillaries in precise registration to ensure uniform spacing between the capillaries, in order to generate uniform hydrodynamic flow in the cuvette. A laser beam excites all of the samples simultaneously, and a microscope objective images fluorescence onto a set of avalanche photodiodes, which operate in the analog mode. A high-gain transimpedance amplifier is used for each photodiode, providing high duty-cycle detection of fluorescence. PMID- 10826678 TI - Separation of polymerase chain reaction amplified bird genes by capillary electrophoresis. AB - Female birds possess one Z and one W chromosome, whereas male birds possess two identical Z chromosomes. Thus, the presence of a W genetic marker is diagnostic of the female sex. Capillary electrophoresis with buffer containing an entangled solution of hydroxyethylcellulose was used to separate the polymerase chain reaction (PCR) amplified bird sexing genes CHD-Z and CHD-W. The relative standard deviations (RSD) were less than 0.6% for the male genes and less than 0.4% for the female genes for six runs and detection limits of 0.1 ng/microL were obtained with laser-induced fluorescence (LIF) detection. Using a DNA ladder and theoretical models for DNA separation in sieving media, the sizes of the two bird genes were determined. PMID- 10826679 TI - Capillary electrophoretic analysis of alkaline phosphatase inhibition by theophylline. AB - An analytical method for studying enzyme inhibition has been developed using capillary electrophoresis with laser-induced fluorescence detection. This technique is based on electrophoretic mixing of zones of enzyme and inhibitor in substrate-filled capillaries. Enzyme catalytic activity is measured by detecting the fluorescent reaction product as it migrates past the detector. Reversible enzyme inhibition is indicated by a transient decrease in product formation. The enzyme, alkaline phosphatase, has been studied using the fluorogenic substrate AttoPhos ([2,2'-bibenzothiazol]-6-hydroxy-benzthiazole phosphate). This assay has been used to quantify theophylline, a noncompetitive, reversible inhibitor of alkaline phosphatase. The detection limit for theophylline is estimated at 3 microM, and 8.6 amole of alkaline phosphatase are required for each assay. The calculated K(i) for theophylline is 90 microM for the capillary electrophoretic enzyme-inhibitor assays. PMID- 10826680 TI - Separation and determination of the macrolide antibiotics (erythromycin, spiramycin and oleandomycin) by capillary electrophoresis coupled with fast reductive voltammetric detection. AB - Separation and determination of erythromycin, spiramycin and oleandomycin by capillary zone electrophoresis coupled with fast reductive voltammetric detection using an Hg-film electrode was investigated in a simple aqueous phosphate buffer system. The influence of pH, concentration of phosphate, applied voltage, capillary length and dimension on the separation was examined and optimized. The entire separation of erythromycin, spiramycin, and oleandomycin was achieved in a 0.2 mol/L phosphate buffer system without organic modifiers. The electrochemical detection parameters, such as electrode material, applied waveform, scan rate, preconcentration potentials and preconcentration times, were investigated and discussed. This approach provides high separation efficiency and high sensitivity for all compounds, with detection limits (3 x peak-to-peak baseline noise) of 7.5 x 10(-8) mol/L for spiramycin, and 3 x 10(-7) mol/ L for erythromycin and oleandomycin. The calibration plot of peak areas for each separated peak vs. concentration of analyte was found to be linear over three orders of magnitude. PMID- 10826681 TI - Design of a sheathless capillary eletrophoresis-mass spectrometry probe for operation with a Z-spray ionization source. AB - The construction of a sheathless interface for capillary electrophoresis electrospray ionization mass spectrometry (CE-ESI-MS), for operation with a Z Spray source on a Micromass Quattro-LC triple quadrupole mass spectrometer is described. Designing the interface involved machining a probe compatible with the setup already in place on the mass spectrometer, i.e., MegaFlow-Z ESI. The probe was made of Lexan with the same dimensions as the ESI probe supplied with the instrument. The electrical connection at the electrospray end of the CE capillary was made possible by gold-coating (sheathless CE-ESI-MS). The probe design as well as the electrical and power supply requirements are described in detail. Experiments were performed using this interface, and CE separations of mixtures containing pmole and sub-pmole amounts of peptides were monitored by on-line MS. For a standard peptide mixture (10(-4) M), separation efficiency was typically characterized by N > 10(4) theoretical plates with S/N > 400. Using the same experimental setup, it was also possible to conduct on-line CE-ESI-tandem MS (MS/MS) experiments on the same peptide mixture, and to determine the sequence of the peptides. MS/MS scan functions for different precursor ions were used either alternately or sequentially and the results from both methods were compared. The possibility of peptide mass mapping was explored, and CE-ESI-MS results were obtained for the digestion products of equine myoglobin. Separation efficiencies and S/N values were similar to those obtained for standard peptides. A complete map of the digestion products was obtained. PMID- 10826682 TI - Subatmospheric electrospray interface for coupling of microcolumn separations with mass spectrometry. AB - A modular subatmospheric electrospray interface with fiber optic UV detection close to the electrospray tip was developed for coupling of microcolumn separation techniques with mass spectrometry. The interface was based on a liquid junction with a removable microelectrospray tip. The electrospray tip was enclosed in a subatmospheric chamber attached in front of the sampling orifice of the mass spectrometer. The inlet of the liquid junction was maintained at atmospheric pressure, and thus no pressure drop developed across the separation column. The flow rate of the electrosprayed liquid from the liquid junction reservoir was adjusted by the pressure in the electrospray chamber. In this approach, a continuous and stable electrospray could be achieved without the use of an external pump. Since the electrospray did not depend on fluid delivery from the separation column, coated capillaries without electroosmotic flow as well as capillaries with electroosmotic flow could be used for capillary electrophoresis. In addition, the interface was found to be effective with capillary liquid chromatography. The use of a fiber optic UV detector placed close to the exit of the separation column provided additional detection information and a simple means of troubleshooting. The interface did not significantly influence the quality of the separation, even with columns generating several hundred thousand theoretical plates. Peptide samples in the submicromolar concentration range were detected, corresponding to a limit of detection in the attomole range. PMID- 10826683 TI - Mass spectrometric detection for capillary isoelectric focusing separations of complex protein mixtures. AB - Capillary isoelectric focusing (CIEF) can provide high-resolution separations of complex protein mixtures, but until recently it has primarily been used with conventional UV detection. This technique would be greatly enhanced by much more information-rich detection methods that can aid in protein characterization. We describe progress in the development of the combination of CIEF with Fourier transform ion cyclotron resonance (FTICR) mass spectrometry and its application to proteome characterization. Studies have revealed 400-1000 putative proteins in the mass range of 2-100 kDa from total injections of approximately 300 ng protein in single CIEF-FTICR analyses of cell lysates for both Escherichia coli (E. coli) and Deinococcus radiodurans (D. radiodurans). We also demonstrate the use of isotope labeling of the cell growth media to improve mass measurement accuracy and provide a means for quantitative proteome-wide measurements of protein expression. The ability to make such comprehensive and precise measurements of differences in protein expression in response to cellular perturbations should provide new insights into complex cellular processes. PMID- 10826684 TI - The identification and determination of selected 1,4-benzodiazepines by an optimised capillary electrophoresis-electrospray mass spectrometric method. AB - An optimised capillary electrophoresis-electrospray mass spectrometric method is presented for the identification and determination of diazepam and its metabolites N'-desmethyldiazepam, oxazepam and temazepam. By investigating constituent parts of the capillary electrophoresis-electrospray mass spectrometric interface and optimising their function, a relatively fast and reproducible method is described for the identification and determination of selected 1,4-benzodiazepines. Optimisation of sheath and auxiliary gas flows, capillary tip tapering, capillary tip positioning, sheath liquid composition and flow rate and pressure application during the separation step have led to acceptable relative standard deviation (RSD) values for migration time and peak area, correlation coefficients and limits of detection. This has been achieved as a result of stabilising the electrospray current prior to analysis, a procedure that takes a matter of minutes when using the method described. Sequential product ion fragmentation (MS(n)) characterisation of 15 1,4-benzodiazepines is also presented and mechanisms for the observed fragmentation patterns proposed. PMID- 10826685 TI - Evaluation of the potentiometric detection of acetylcholine and other neurotransmitters in capillary electrophoresis. AB - It is demonstrated that acetylcholine may be determined potentiometrically in capillary electrophoresis down to 6 x 10(-7) M with a miniature coated-wire ion selective electrode. The more established amperometric method is not possible for this compound. Similarly, 4-aminobutyric acid (GABA) is not accessible amperometrically; although potentiometric detection was possible, the sensitivity is not good. Other neurotransmitters may be determined by either method, but in these cases amperometric determination is generally favorable. For serotonin the potentiometric detection limit was 4 x 10(-5) M, while for the amperometric method a detection limit of 10(-7) M was found. PMID- 10826686 TI - In-line coupling capillary electrochromatography with amperometric detection for analysis of explosive compounds. AB - Amperometric detection at a bare gold electrode has been in-line coupled with capillary electrochromatography (CEC) for analysis of nitroaromatic and nitroamine explosives in contaminated soils and ground water. The CEC column packed with 3 microm C18 particles performed best using a mobile phase containing 70-80% methanol, 30 or 20% water, 5 mM sodium dodecyl sulfate (SDS) and 10mM 2-(N morpholino)ethanesulfonic acid (MES). In contrast, the separation column packed with 1.5 km C18 particles exhibited the best separation when only 30% methanol was added to a mobile phase containing 70% water, 7 mM SDS, and 10 mM MES. The detection, based on electrochemical reduction of the explosives (-0.7 or -1 V vs. Ag/AgCl, depending upon the level of methanol in the mobile phase), was compatible with such mobile phases. The detection limits for 13 explosives ranged from 100 to 200 ppb, i.e., about twofold better than those obtained with electrokinetic chromatography (EKC)/amperometric detection. From an operational viewpoint, exhaustive column conditioning was a prerequisite and care should be taken to prevent bubble formation and current breakdown during the course of separation. The CEC column equipped with amperometric detection successfully measured explosives in ground water and extracts prepared from contaminated soils and the results obtained agreed well with those of the U.S. Environmental protection Agency (EPA) method. PMID- 10826687 TI - Conductivity detection of aliphatic alcohols in micellar electrokinetic chromatography using an oscillometric detector. AB - Although conductivity is usually applied to detect ionic species in capillary electrophoresis (CE), nonionic species can also be detected by their indirect effects on the conductivity of the running electrolyte. This approach was used for detection of aliphatic alcohols in micellar electrokinetic chromatography (MEKC) with an oscillometric detector. Although the detector operates at 600 kHz, for the range of electrolyte concentration used in CE, the response is mainly due to variations of conductivity. A 50 mM phosphate and 50 mM SDS solution was used as running electrolyte and as the solvent for mixtures of some isomers of propanol, butanol, and pentanol. A set of negative peaks was obtained and assigned to the components by spiking the samples. The limits of detection (LOD) ranged from 2.1 mM for 2-methyl-2-propanol to 5.3 mM for 1-pentanol. Due to the high affinity for the interior of the micelles, 1-hexanol could not be easily detected, but by the addition of 10% methanol to the running electrolyte it was possible. For this electrolyte, the LOD was improved, ranging from 0.8 mM for 2 methyl-2-propanol to 1.5 mM for 1-pentanol. Calibration plots were linear up to 40 mM at least. These results indicate that conductivity may be useful for detection of nonionic species in CE, especially when optical methods can not be conveniently applied. PMID- 10826688 TI - Quantitative assay of metronidazole by capillary zone electrophoresis with amperometric detection at a gold microelectrode. AB - Capillary zone electrophoresis was employed for the determination of metronidazole using end-column amperometric detection with a gold microelectrode at a constant potential of -0.52V vs. saturated calomel electrode. To overcome interference of oxygen in the solution, a deaeration injector and a deaeration protector at the detection cell were used. The optimum conditions of separation and detection are 1.0 x 10(-3) mol/L potassium dihydrogen citrate (KH2C6H5O7) for the buffer solution, 20 kV for the separation voltage, and 5 kV and 10 S for injection voltage and injection time, respectively. The limit of detection is 6.0 x 10(7) mol/L or 0.78 fmole (S/N = 3). The relative standard deviation is 3.9% for the electrophoretic peak current. The method was applied to the determination of metronidazole in human urine. PMID- 10826689 TI - Determination of clozapine by capillary zone electrophoresis following end-column amperometric detection with simplified capillary/electrode alignment. AB - Capillary zone electrophoresis was employed for the determination of clozapine using an end-column amperometric detection at a carbon fiber array microdisk electrode with simplified capillary/electrode alignment. The optimum conditions of separation and detection are: Britton-Robinson buffer, pH 2.0 (1.3 x 10(-2) mol/L total concentration of acids, 3.2 x 10(-3) mol/L NaOH), 15 kV for separation voltage, 5 kV and 10 s for injection voltage and injection time, respectively. The limit of detection is 4.2 x 10(-7) mol/L or 1.2 fmole (signal to noise, S/N = 2). The relative standard deviation is 1.4% for the migration time and 2.5% for the electrophoretic peak current. The method was applied to the determination of clozapine in human blood. The recovery of the method is between 94-104%. PMID- 10826690 TI - Parameters affecting reproducibility in capillary electrophoresis. AB - It is well known that poor quantitative reproducibility substantially limits the practical implementation of capillary electrophoresis (CE) separations in chemical analysis. The principal sources of variance in observed peak areas are irreproducible flow rate, which influences on-column detector response, and inconsistent injection volume or amount. An overview of studies by researchers to address the reproducibility issue will be presented. In addition, current efforts in our laboratory to assess sources of quantitative variance for separations of dansylated amino acids using an automated CE system are presented and related when appropriate to the body of existing knowledge on this important topic. A comparison of different injection methods (hydrostatic vs. electrokinetic) and approaches (e.g., high vs. low pressure), the effect of random changes in electroosmotic flow (EOF) due to air bubbles in the CE capillary, and choice of certain peak integration parameters in terms of peak area reproducibility are presented. Under optimum conditions relative standard deviation (RSD) values in raw peak area are typically 2.0%. With nonoptimum conditions (e.g., with air bubbles in capillary), RSD values can substantially degrade. However, normalizing with retention times, internal standards, or observed electrophoretic current produces RSD values in a range of 1.4-2.3%. PMID- 10826691 TI - Softening of fused-silica capillaries during particle packing. AB - When a semipreparative capillary electrochromatography (CEC) capillary is packed with silica particles and exposed to solvent, its mechanical strength is markedly reduced. In our studies, a fused-silica capillary (internal diameter > 200 microm and wall thickness < 150 microm) was packed under pressure (approximately 200 psi) with spherical silica particles (1.5-5 microm) suspended in water or various common organic solvents. After one hour of exposure, the capillary can be readily deformed, and it keeps its deformed shape upon release of the force causing deformation. It is suggested that capillary softening is promoted through the propagation of internal microcracks that have been caused by action of the particles during packing in the presence of solvent. Application of a protective coating to the inside of the capillary is found to reduce or eliminate capillary softening. PMID- 10826692 TI - Contiguous strings of strongly binding short oligonucleotides as a useful tool for completing sequencing experiments. AB - Large-scale DNA sequencing is currently based mostly on the shotgun approach. Although widely used, it is not free of shortcomings: the total length of randomly sequenced subclones appear to be five to eight times more than the total DNA length; such a high statistical redundancy of random sequences cannot guarantee the absence of unsequenced gaps. We calculated that these problems could be alleviated if the average length of the subcloned random fragments were increased. In this paper we present the primer walking approach based on the use of contiguous modified hexamer strings strongly binding to complementary templates as segmented primers. The approach exhibits the following characteristic features: 1) Single-stranded templates appear to have advantage over double-stranded ones. 2) With single-stranded templates, an overall sequencing success rate is about 80%. 3) A relatively small library of 1400 hexanucleotides selected according to the most frequent occurrence in the human genome is sufficient for sequencing all clones. The approach allows one to reduce the number of templates necessary for the shotgun strategy and also to decrease the number of gaps between sequenced contigs. This should be kept in mind when considering the ways for enhancement of the present strategy of large-scale sequencing. PMID- 10826693 TI - Cloning and sequencing of the genes encoding the periplasmic-cytochrome B containing selenate reductase of Thauera selenatis. AB - The periplasmic selenate reductase (Ser) of Thauera selennatis is a component of the electron transport chain catalyzing selenate reduction with acetate as the electron donor (i.e., selenate respiration). The purified enzyme consists of three subunits (SerA, SerB and SerC). Using transposon (i.e., Tn5) mutagenesis selenate reductase mutants were isolated. Junction fragments of DNA adjacent to the integrated Tn5 were used, together with oligonucleotides derived from the N termini of SerA and SerB, to clone from a gene bank a DNA fragment that contained the corresponding genes. After sequencing, serA, serB and serC were identified by sequence comparison with the N-termini of the three subunits. The genes are arranged in the order serA, serB, serC; a fourth open reading frame (serD) in between, but overlapping serB and serC, is also present. The serA gene product contains an apparent leader peptide with a twin-arginine motif. The remainder of the translated amino acid sequence is similar to that of a number of prokaryotic molybdenum-containing enzymes (e.g., nitrate reductases and formate dehydrogenases of Escherichia coli). The serB gene product contains four cysteine clusters and is similar to various iron-sulfur protein subunits. The serC gene product contains a putative Sec-dependent leader peptide, but there are no similarities between the remainder of the translated protein and other protein subunits. The SerC contains two histidine and four methionine residues, and these may noncovalently bind heme b--which is a component of the active selenate reductase. The serD gene product encodes a putative protein that shows no significant sequence similarities to other proteins. However, the location of the serD within the other ser genes is similar to that of narJ within the E. coli narGHJI operon (nitrate reductase A); thus suggesting that the role of SerD may be similar to that of NarJ, which is a system-specific chaperone protein. PMID- 10826694 TI - Structural analysis and transcript processing of the bovine proteolipid protein (PLP) gene. AB - In this study we present the complete genomic structure of the bovine PLP gene and its assignment to the long arm of the X-chromosome (BTXq2.1). We determined a total of 18,767 bp of the bovine PLP gene and compared it to the human heterolog. A very high similarity was detected between the non-coding regions, interrupted primarily by several transposable elements. A deletion of 13 bp in the vicinity to the translation start signal in the promoter of the bovine PLP gene was found. Functional studies of the 3' region showed the use of several polyadenylation signals. Three main transcripts were detected in adult cattle in the range of 3200, 2400, and 1600 nucleotides using Northern blot analysis. An additional shorter transcript was detected in the cerebrum of calves. PMID- 10826695 TI - Cloning and sequencing of feline and canine ice-related cDNAs encoding hybrid caspase-1/caspase-13-like propeptides. AB - Caspases are cysteine proteases which have important roles in the activation of cytokines and in apoptosis. The ICE subfamily of caspases comprise peptides closely related to caspase-1, or interleukin-1beta (IL-1beta) converting enzyme (ICE), which promotes maturation of interleukin IL-1beta and interleukin-18 (IL 18) by proteolytic cleavage of precursor forms to generate biologically active peptides. Other members of this subfamily include caspase-4, -5, -13 and isoforms of these proteins. We report the cloning and sequencing of two feline and canine ICE-related cDNAs amplified by RT-PCR. The predicted proteins are 410 and 404 amino acids in length respectively and are most closely related to caspase-1 sequences across the N-terminal 115 amino acids and to human caspase-13 across the C-terminal sequence. PMID- 10826696 TI - Isolation and expression of a barley beta-1,3-glucanase isoenzyme II gene. AB - A beta-1,3-glucanase gene from Hordeum vulgare was isolated by a PCR strategy, cloned and subsequently sequenced. The amplified sequence contained the entire coding region of the isoenzyme II, which is interrupted by a 165 bp intron at 73 bp downstream the starting codon. This intron contains all the elements required for the processing mechanism in monocots: a high A + U content, the appropriate splice sites in the 5' and 3' ends and four typical YUNAN consensus sequences. Transient transformation of wheat protoplasts with the complete beta-1,3 glucanase gene under the control of maize polyubiquitin promoter revealed that the intron sequence was spliced out. The gene was also expressed at high levels, probably due to an enhancer-like sequence found near the 3' end of the intron. PMID- 10826697 TI - cDNA cloning of lymphotoxin alpha (LT-alpha) from a marsupial, Macropus eugenii. AB - Lymphotoxin (LT) is a proinflammatory cytokine with a broad spectrum of immunological activities. While the 'classic' form of the molecule is a secreted homotrimer, now referred to in the literature as LT-alpha3, it has more recently been recognised that a membrane-bound form of LT exists on activated T lymphocytes and that this represents a complex between LT-alpha and a closely related type II membrane protein, LT-beta. Together with another related cytokine, tumour necrosis factor alpha (TNF-alpha), these molecules have been extremely well studied in eutherian mammals but not in any other group. Marsupials represent a distinct branch in mammalian evolution to that of eutherians, the two groups having diverged more than 100 million years ago. We report here for the first time, the cDNA cloning of LT-alpha from a marsupial, Macropus eugenii (tammar wallaby). This sequence was found to be relatively conserved when compared to orthologous sequences from eutherian mammals, sharing an average sequence identity of 70.4% at the nucleotide level and 71.7% at the deduced amino acid level. PMID- 10826698 TI - Nucleotide sequence of the rat CD40 ligand. PMID- 10826699 TI - TMpcp: a Tuber magnatum gene which encodes a putative mitochondrial phosphate carrier. AB - Little is known about the genome of Tuber, Ascomycetes which comprise a number of ectomycorrhizal species. Screening of a genomic library of Tuber magnatum led to identification of a chitin synthase gene (chs). On sequencing upstream of it in the same phage, we found a 2000 bp long fragment that proved to contain a hypothetical gene with high homology with mitochondrial phosphate carriers from human and bovine heart, and from Saccharomyces cerevisiae. The sequence contains two putative introns and its open reading frame encodes for a protein 305 amino acids long. A primary sequence analysis revealed 6 hydrophobic segments and a signature pattern, similar to that of other mitochondrial carriers. PMID- 10826700 TI - Identification of a novel thioredoxin-1 pseudogene on human chromosome 10. PMID- 10826701 TI - Genomic structure of mouse and human genes for DNA-PKcs interacting protein (KIP). AB - DNA-dependent protein kinase (DNA-PK) is a nuclear protein serine/threonine kinase in a wide variety of vertebrate species and it has a role in the DNA repair and recombination process of lymphoid development. DNA-PK is composed of a large catalytic subunit (DNA-PKcs) and DNA-binding protein, Ku. Recently, the mouse and human DNA-PKcs interacting proteins (Kip/KIP) have been reported. In this report, we have determined the complete genomic structure of mouse and human Kip/KIP genes. The total length of mouse Kip gene and human KIP gene are approximately 5.7 kb and 3.6 kb in genomic DNAs, respectively. Both of genes consist of 7 exons. PMID- 10826702 TI - Genomic structure and chromosomal localization of human thioredoxin-like protein gene (txl). AB - Human thioredoxin-like protein (txl) is a novel member of the expanding thioredoxin superfamily whose main characteristic is the presence, after the thioredoxin domain, of a C-terminal extension of 184 residues with no homology with any other protein in the databases. Txl is a cytosolic ubiquitously expressed protein and it has been copurified with a kinase of the STE20 family, which is proteolytically activated by caspases in apoptosis. However, no cellular function has yet been assigned to this protein. In the present study we report the genomic organization of the txl gene which encompasses approximately 36 kb organized in eight exons ranging from 96 bp to 303 bp. In contrast, intron sizes are much bigger ranging from 1.5 kb to 12 kb. Chromosomal localization of txl gene revealed that it maps at position 18q21, a region frequently affected in different human tumours. Furthermore, we have identified the putative homologues of txl in both Drosophila melanogaster and Caenorhabditis elegans that display much closer homology to the known thioredoxins than the human txl protein. Indeed, critical residues for optimal thioredoxin activity are present in both Drosophila and Caenorhabditis txl but absent in the human protein suggesting that txl might have evolved to carry out a function different from the general disulfide reductase typical of thioredoxins. PMID- 10826703 TI - Cloning, sequence and mRNA expression of bovine interleukin-16. AB - A lambda gt11 cDNA library was constructed using mRNA isolated from Theileria parva-infected bovine lymphocytes. Sequencing of random clones of this library resulted in the identification of a cDNA encoding bovine interleukin-16 (IL-16). The cDNA has an open reading frame of 1134 bp, and a 3' untranslated region of 275 nucleotides with a polyadenylation signal 16 bases upstream from the poly (A) tail. The protein predicted by the cDNA sequence contains 378 amino acids and the level of amino acid homology with the corresponding part of human precursor IL-16 is 79 %. No information is available about the tissue distribution of IL-16 in cattle, therefore we investigated the expression of IL-16 mRNA in bovine lymphoid tissues by reverse-transcription polymerase chain reaction assays. To investigate the potential of IL-16 as an immunoregulatory molecule we also analysed IL-16 mRNA expression in CD4+ and CD8+T-cell clones derived from T. parva-immunised cattle. PMID- 10826704 TI - Full promoter sequence of human early growth response factor-1 (Egr-1): demonstration of a fifth functional serum response element. AB - The early growth response factor-1 (Egr-1) gene encodes a zinc finger transcription factor which is critical for cell proliferation and differentiation. The human Egr-1 promoter comprises regulatory elements including two Sp1 sites, an AP1 site, two cAMP response elements and an Egr-1 binding site. In addition to these transcription factor binding sites, the promoter harbours five serum response elements (SREs) and associated binding sites for the Ets transcription factor family, previously identified from partial sequence data (Sakamoto et al, Oncogene 6; 867-871, 1991). We now report the full sequence of the human Egr-1 promoter and confirm the presence of a fifth serum response element. This element is functionally active in a minimal promoter vector in response to the MAP kinase kinase MEK1. PMID- 10826705 TI - Protein crosslinking in assembly and remodelling of extracellular matrices: the role of transglutaminases. AB - Transglutaminases form a family of proteins that have evolved for specialized functions such as protein crosslinking in haemostasis, semen coagulation, or keratinocyte cornified envelope formation. In contrast to the other members of this protein family, tissue transglutaminase is a multifunctional enzyme apparently involved in very disparate biological processes. By virtue of its reciprocal Ca2+-dependent crosslinking activity or GTP-dependent signal transducing activity, tissue transglutaminase exhibits true multifunctionality at the molecular level. The crosslinking activity can subserve disparate biological phenomena depending on the location of the target proteins. Intracellular activation of tissue transglutaminase can give rise to crosslinked protein envelopes in apoptotic cells, whereas extracellular activation contributes to stabilization of the extracellular matrix and promotes cell-substrate interaction. While tissue transglutaminase synthesis and activation is normally part of a protective cellular response contributing to tissue homeostasis, the enzyme has also been implicated in a number of pathological conditions including fibrosis, atherosclerosis, neurodegenerative diseases, celiac disease, and cancer metastasis. This review discusses the role of transglutaminases in extracellular matrix crosslinking with a focus on the multifunctional enzyme tissue transglutaminase. PMID- 10826706 TI - The strain magnitude and contact guidance determine orientation response of fibroblasts to cyclic substrate strains. AB - When grown in a substrate subjected to cyclic stretching, most types of cells change orientations. This cell orientation response (COR) has been shown to be driven by axial substrate strain (the strain beneath and along a cell's long axis). However, it remains unclear whether COR depends on the strain direction (tension vs compression). Furthermore, in vitro COR is paradoxical, since in vivo fibroblasts align along collagen fibers and hence the stretch direction. We hypothesized that COR does not depend on the surface strain direction, and that contact guidance provided by microgrooves can maintain cell alignment in the presence of cyclic stretching. Human skin fibroblasts were cultured on compliant smooth and microgrooved surfaces in silicone dishes. Cyclic uniaxial tensile and compressive strains (4%, 8% and 12%) were applied on the dishes at 1 Hz for 24 h. Cell orientation distributions were determined and compared using the Kolmogorov Smirnov test. Significant differences were found between each of cell orientation distributions with the applied strains and that without strains (p < 0.05). Nevertheless, no significant differences were found between two cell orientation distributions for each pair of opposite strains applied (for 4%, p = 0.33; for 8%, p = 0.18; and for 12%, p = 0.32). Moreover, fibroblasts grown in microgrooves aligned in the groove direction and remained so after 8% cyclic stretch. Thus, this study showed that COR is the cells' avoidance to substrate deformation (i.e., strain-direction independent). It also suggested that the failure of fibroblasts to change orientations in vivo may result from the contact guidance provided by collagen fibers. PMID- 10826707 TI - Structural aspects of the calcification process of lower vertebrate collagen. AB - In order to investigate the structural relationship between inorganic phase and collagen fibrils in the calcified tissues of lower vertebrates we have carried out a wide and small angle X-ray diffraction investigation on carp scales and bone samples. The small angle patterns from decalcified bone and scales, as well as uncalcified tendon samples from carp are very similar to that of type I collagen from higher vertebrates. The D-axial period, 67 nm, is the same as that of higher vertebrate type I collagen, while the most significant difference is the relatively low intensity of the first order reflection, which is, however, the most intense. The relative intensity distributions of the meridional reflections recorded from fish bone and scales are in agreement with an electron density distribution according to a step function. The calculated step length is very close to the values previously reported for calcified tissues from higher vertebrates. The small angle reflections from calcified, as well as decalcified, scales display different directions of orientation, which could be in agreement with a plywood arrangement of collagen fibrils in successive sheets parallel to the plane of the scale. PMID- 10826708 TI - Pluripotential marrow cells produce adipocytes when transplanted into steroid treated mice. AB - The effect of steroids on adipogenesis by D1-BAG, a pluripotent cell cloned from mouse bone marrow and transfected with traceable genes encoding beta galactosidase and neomycin resistance, was investigated in vitro in culture and in vivo after injection into mice. Treatment of D1-BAG cells in culture with dexamethasone produced an accumulation of lipid vesicles and stimulated expression of the fat cell-specific 422(aP2) mRNA. Fifty-six mice each received 1 x 10(6) D1-BAG cells, either by tail-vein injection or by direct injection into the marrow of the right femur. Another 38 mice received either saline injection or no treatment as controls. Half of the animals in each group were treated with 3 mg/kg of methylprednisolone per week. Analysis of marrow blow-outs by flow cytometry, DNA analysis by PCR, and X-gal stain of histological sections indicated that cells transplanted by either intravenous or intramedullary injection had appeared and persisted in the marrow of host mice. Cell sorting by flow cytometry and staining with Sudan IV demonstrated that steroid treatment produced adipogenesis in 5-9% of transplanted cells. The results indicate that steroid-induced differentiation of potentially osteogenic marrow cells into adipocytes in vivo may contribute to the development of osteoporosis and osteonecrosis. PMID- 10826709 TI - Cell surface proteoglycan expression by human periodontal cells. AB - Cell surface proteoglycans are known to be involved in many functions including interactions with components of the extracellular microenvironment and serve to influence cell shape, adhesion, proliferation, and differentiation. They also can act as co-receptors, to help bind and modify the action of various growth factors and cytokines. Despite their strategic location and relevance to cell function, few studies have considered the nature of the cell surface proteoglycans associated with cells of the periodontium. Due to the structural complexity and multiplicity of cell types in the periodontium, we have selected three different cell lines (gingival connective tissue fibroblast, periodontal ligament fibroblast, and osteoblast) which each represent the unique functions within the periodontium to study the expression of cell surface proteoglycans. We hypothesized that a number of cell surface proteoglycans will be expressed by human periodontal cells and these may be related to the source and function of the cell. Western blotting and RT-PCR methods were used to study the expression of five cell surface proteoglycans (syndecan-1, -2, -4, glypican and betaglycan) in three cell lines of human periodontal cells in vitro. Our results demonstrated the expression of protein cores for syndecan-1 (43 kDa), syndecan-2 (48 kDa), syndecan-4 (35 kDa), glypican (64 kDa), and betaglycan (100-110 kDa). RT-PCR results confirmed that all of these cells produced mRNA for the cell surface proteoglycans under study, of which syndecan-2 showed a significant difference in expression between the periodontal ligament fibroblasts, gingival fibroblasts and osteoblasts. We conclude that the presence of specific cell surface proteoglycans on periodontal cells implies a likely role for these molecules in cell-cell, cell matrix interactions involved in periodontal disease and/or regeneration of the periodontium, of which they may have distinctive functions related to the source and function of these cells. PMID- 10826710 TI - Effect of nicotine on rat gingival fibroblasts in vitro. AB - Tobacco smoking is considered a major risk factor for the development and progression of periodontal diseases (Haber, J. and Wattles, J. (1994). J. Periodontol., 64, 16-23). The purpose of this study was to determine the effects of nicotine on rat gingival fibroblasts (RGF) cultured in vitro. After ether anesthesia, rat gingival tissues were obtained from the attached gingiva of a Wistar rat. Small fragments of gingiva were maintained in culture in Petri dishes. Fibroblasts developing from these explants were collected to obtain monolayer cultures. After the fourth passage (T4), cells were supplemented with nicotine at various concentrations. Control and treated cells were examined under phase contrast or transmission electron microscopy. They were compared as regards their DNA content, mitochondrial activity, collagen and protein synthesis, and cell death by apoptosis or necrosis. Nicotine from 0.05 microM to 1 mM did not affect the DNA content or protein and collagen synthesis. At concentrations between 3 and 5 mM, growth was significantly diminished and the survival rate reduced. Ultrastructural analysis revealed dilated mitochondria and vacuolization in treated cells, suggestive of necrosis, but increased apoptosis was also revealed by cytometry. On the basis of this in vitro study, it appears that tobacco, through its component nicotine, may directly affect various functions of RGF. PMID- 10826711 TI - Efficacy of ultraviolet light for reducing Escherichia coli O157:H7 in unpasteurized apple cider. AB - This study examined the efficacy of UV light for reducing Escherichia coli O157:H7 in unpasteurized cider. Cider containing a mixture of acid-resistant E. coli O157:H7 (6.3 log CFU/ml) was treated using a thin-film UV disinfection unit at 254 nm. Dosages ranged from 9,402 to 61,005 microW-s/cm2. Treatment significantly reduced E. coli O157:H7 (P < or = 0.0001). Mean reduction for all treated samples was 3.81 log CFU/ml. Reduction was also affected by the level of background microflora in cider. Results indicate that UV light is effective for reducing this pathogen in cider. However, with the dosages used in this experiment, additional reduction measures are necessary to achieve the required 5 log reduction. PMID- 10826712 TI - Fate of Shigella sonnei on parsley and methods of disinfection. AB - Outbreaks of shigellosis associated with chopped parsley used as a garnish for foods occurred in four states in the United States and in two Canadian provinces in 1998. This prompted a study to determine survival and growth characteristics of Shigella sonnei inoculated onto raw parsley. Two inoculum levels (approximately 10(3) and 10(6) CFU/g) were applied to parsley leaves, portions of which were then chopped. Inoculated whole and chopped parsley leaves were held at 4 degrees C or 21 degrees C for up to 14 days. Initial populations of the organism on chopped parsley receiving high or low levels of inoculum increased by approximately 3 log10 CFU/g, within 1 day at 21 degrees C. Populations of S. sonnei on inoculated chopped or whole parsley leaves held at 4 degrees C decreased by 2.5 to 3.0 log10 CFU/g during a 14-day storage period. The pathogen multiplied, without a lag phase, on inoculated (2.72 log10 CFU/g) chopped parsley held at 21 degrees C, exceeding 6 log10 CFU/g within 24 h. Treatment of inoculated whole parsley leaves with vinegar containing 5.2% (vol/vol) acetic acid or 200 ppm free chlorine for 5 min at 21 degrees C reduced the population of S. sonnei by more than 6 log10 CFU/g, whereas treatment with vinegar containing 7.6% acetic acid or 250 ppm free chlorine reduced initial populations of 7.07 and 7.26 log10 CFU/g, respectively, to undetectable levels (<0.6 log10 CFU/g). These studies revealed that S. sonnei can grow rapidly on chopped parsley held at ambient temperature and remain viable for at least 14 days at 4 degrees C. Treatment of contaminated parsley with vinegar or chlorinated water offers a simple method to reduce markedly or eliminate the pathogen in food-service or home settings. PMID- 10826713 TI - Short-chain volatile fatty acids modulate the expression of the hilA and invF genes of Salmonella typhimurium. AB - The ability of Salmonella typhimurium to invade the intestinal mucosal cells is an important step in pathogenesis. This invasion process requires genes encoded on the Salmonella pathogenicity island 1 (SPI1). Two transcriptional activators, HilA and InvF, encoded in SPII regulate the expression of invasion genes in response to environmental stimuli such as osmolarity, oxygen tension, and pH. During its pathogenic life cycle, Salmonella typhimurium is also exposed to short chain fatty acids (SCFA), especially acetate, propionate, and butyrate, in the intestinal lumen, as well as the SCFA used as food preservatives. The effects of SCFA on the expression of hilA and invF-lacZY transcriptional fusions were examined to determine the potential role of SCFA in the pathogenesis of Salmonella typhimurium. Growth rates were reduced by increasing SCFA concentrations at pH 6 but not at pH 7. At pH 7, hilA and invF expression was induced by acetate but not by propionate or butyrate, while at pH 6, all SCFA induced hilA and invF expression at 1 h. In general, hilA and invF expression levels when compared to respective control responses were higher at 1 h than at 4 and 8 h in the presence of most SCFA concentrations at pH 6. However, expression levels at 4 and 8 h were either similar or higher than the 1-h responses for the hilA-lacZY fusion strain in the presence of acetate while exposure to 20 mM propionate yielded similar levels of expression at 1, 4, and 8 h. The pH dependent manner of induction suggests that entry of SCFA into the cell was necessary for induction. We speculate that SCFA may serve as an environmental signal that triggers the expression of invasion genes in the gastrointestinal tract. PMID- 10826714 TI - Incidence of Salmonella in fish and seafood. AB - Field laboratories of the U.S. Food and Drug Administration collected and tested 11,312 import and 768 domestic seafood samples over a 9-year period (1990 to 1998) for the presence of Salmonella. The overall incidence of Salmonella was 7.2% for import and 1.3% for domestic seafood. Nearly 10% of import and 2.8% of domestic raw seafood were positive for Salmonella. The overall incidence of Salmonella in ready-to-eat seafood and shellfish eaten raw was 0.47% for domestic -one shucked oyster and one shark cartilage powder. The incidence in the 2,734 ready-to-eat import seafood was 2.6%--cooked shrimp, shellfish or fish paste, smoked fish, salted/dried fish, and caviar. The incidence in import shellfish consumed raw was 1% in oyster, 3.4% in clams, and 0% in mussels. The incidence in raw, import fish was 12.2%. Distribution of Salmonella in seafood on a regional basis indicated the incidence to be highest in central Pacific and Africa and lowest in Europe/Russia and North America (12% versus 1.6%). Data on a country basis indicated Vietnam to have the highest (30%) and Republic of Korea the lowest (0.7%). While the most frequent serotypes in import seafood were Salmonella Weltevreden (1st), Salmonella Senftenberg (2nd), Salmonella Lexington, and Salmonella Paratyphi-B (3rd, equal numbers for each serotype), the top 20 list included Salmonella enteritidis (5th), Salmonella Newport (6th), Salmonella Thompson (7th), Salmonella typhimurium (12th), and Salmonella anatum (13th), commonly involved in foodborne illness in the United States. Because the incidence in the present study is based on only a small fraction of the seafood imported into the United States, efforts should be directed toward implementation of hazard analysis and critical control points to reduce the incidence of Salmonella in seafood without relying on testing for Salmonella. PMID- 10826715 TI - Antimicrobial activity of cetylpyridinium chloride washes against pathogenic bacteria on beef surfaces. AB - Cetylpyridinium chloride (CPC), a water-soluble, neutral pH, colorless compound, is widely used in oral hygiene products to inhibit bacteria responsible for plaque. Previously, researchers have demonstrated that CPC not only reduces Salmonella typhimurium on poultry but also prevents cross-contamination. To determine the effectiveness of CPC against pathogens associated with lean and adipose beef surfaces, several spray-washing experiments (862 kPa, 15 s, 35 degrees C) with 1% (wt/vol) CPC were conducted. On lean beef surfaces, CPC immediately reduced 5 to 6 log10 CFU/cm2 of Escherichia coli O157:H7 and Salmonella typhimurium to virtually undetectable levels (0 log10 CFU/cm2), as well as after 35 days of refrigerated (4 degrees C), vacuum-packaged storage. On adipose beef surfaces, 5 log10 CFU/cm2 Salmonella typhimurium and E. coli O157:H7 were reduced immediately (>2.5 log10 CFU/cm2) with 1% CPC; by day 35 the reduction was <1.3 log10 CFU/cm2. Further plate overlay analyses indicated that the effectiveness of CPC against pathogens on adipose surfaces was not hampered by the presence of meat components or fatty acids. Additional chemical and microbiological analyses of 1% CPC-treated beef surfaces subjected to a secondary water wash (following contact times of 0, 5, 10, 15, or 30 min) or grinding did reduce pathogenic bacteria and CPC levels. However, residual CPC levels following any of the treatments were considered excessive for human consumption. Despite the residual levels, this study is the first to demonstrate the effect of CPC on pathogenic bacteria associated with beef surfaces immediately after treatment and also after long-term, refrigerated, vacuum-packaged storage. PMID- 10826716 TI - Antimicrobial effect of herb extracts against Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella typhimurium associated with beef. AB - The effects of plant extracts against pathogenic bacteria in vitro are well known, yet few studies have addressed the effects of these compounds against pathogens associated with muscle foods. A series of experiments was conducted to determine the effectiveness of a commercially available, generally recognized as safe, herb extract dispersed in sodium citrate (Protecta One) or sodium chloride (Protecta Two) against Escherichia coli O157:H7, Salmonella typhimurium, and Listeria monocytogenes associated with beef. In the first experiment, E. coli O157:H7, Salmonella typhimurium, and L. monocytogenes inoculated onto beef and subjected to surface spray treatments with 2.5% solutions of Protecta One or Protecta Two were not affected by immediate application (day 0) of the herbal extracts. However, after 7 days of storage at 4 degrees C, E. coli O157:H7 was reduced by >1.3 log10 CFU/cm2 by Protecta Two; L. monocytogenes was reduced by 1.8 and 1.9 log10 CFU/cm2 by Protecta One and Protecta Two, respectively; Salmonella typhimurium was not reduced >0.3 log10 CFU/cm2 by either extract by day 7. In the second experiment, 2.5% Protecta Two (wt/vol or wt/wt) added to inoculated lean and adipose beef trim, processed, and packaged as ground beef chubs (80% lean, 20% adipose), did not reduce pathogen populations >0.5 log10 CFU/g up to 14 days at 4 degrees C. In the third experiment, surface spray treatments of beef with 2.5% lactic acid or 2.5% solutions of Protecta One or Protecta Two, vacuum packaged, and stored up to 35 days at 4 degrees C did reduce E. coli O157:H7, L. monocytogenes, and Salmonella Typhimurium slightly. These studies suggest that the use of herb extracts may afford some reductions of pathogens on beef surfaces; however, the antimicrobial activity may be diminished in ground beef by adipose components. PMID- 10826717 TI - Fate of Listeria monocytogenes, Salmonella typhimurium DT104, and Escherichia coli O157:H7 in Labneh as a pre- and postfermentation contaminant. AB - Commercially pasteurized milk (approximately 2% milkfat) was heated at 85 to 87 degrees C/30 min, inoculated to contain 2,000 to 6,000 CFU/ml of Listeria monocytogenes, Salmonella typhimurium DT104, or Escherichia coli O157:H7, cultured at 43 degrees C for 4 h with a 2.0% (wt/wt) commercial yogurt starter culture, stored 12 to 14 h at 6 degrees C, and centrifuged to obtain a Labneh like product. Alternatively, traditional salted and unsalted Labneh was prepared using a 3.0% (wt/wt) starter culture inoculum, similarly inoculated after manufacture with the aforementioned pathogens, and stored at 6 degrees C and 20 degrees C. Throughout fermentation, Listeria populations remained unchanged, whereas numbers of Salmonella increased 0.33 to 0.47 logs during the first 2 h of fermentation and decreased thereafter. E. coli populations increased 0.46 to 1.19 logs during fermentation and remained that these levels during overnight cold storage. When unsalted and salted Labneh were inoculated after manufacture, Salmonella populations decreased >2 logs in all samples after 2 days, regardless of storage temperature, with the pathogen no longer detected in 4-day-old samples. Numbers of L. monocytogenes decreased from 2.48 to 3.70 to < 1.00 to 1.95 logs after 2 days with the pathogen persisting up to 15 days in one lot of salted/unsalted Labneh stored at 6 degrees C. E. coli O157:H7 populations decreased from 3.39 to 3.7 to < 1.00 to 2.08 logs during the first 2 days, with the pathogen no longer detected in any 4-day-old samples. Inactivation rates for all three pathogens in Labneh were unrelated to storage temperature or salt content. Unlike L. monocytogenes that persisted up to 15 days in Labneh, rapid inactivation of Salmonella typhimurium DT104 and E. coli O157:H7 suggests that these emerging foodborne pathogens are of less public health concern in traditional Labneh. PMID- 10826718 TI - Pathogenicity and production of virulence factors by Listeria monocytogenes isolates from channel catfish. AB - Pathogenicity of four channel catfish Listeria monocytogenes isolates (CCF1, CCF4, HCC7, and HCC23) was examined in a comparative manner with virulent type strains L. monocytogenes ATCC 19115 and EGD and avirulent type strain ATCC 15313 in BDF and A/J mice. Isolates HCC7 and CCF1 (both serovar 1) caused similar percent mortalities and 50% lethal concentration values when compared with virulent type strains and were therefore considered pathogenic. The presence of the virulence factors listeriolysin (LLO), phosphotidylcholine-phospholipase (PC PLC), and phosphotidylinositol-phospholipase (PI-PLC) was determined using specific activity tests. The virulent catfish isolates were positive for production of LLO, PC-PLC, and PI-PLC. However, catfish isolate HCC23 was not virulent in mice despite being hemolytic, suggesting that not every hemolytic L. monocytogenes strain is virulent. With the exception of HCC7, all virulent strains displayed enhanced LLO production in a special stress medium, whereas almost undetectable LLO activity was present when catfish isolates and virulent type strain L. monocytogenes were grown in a rich medium such as brain heart infusion. Avirulent strains were found to lack or have decreased expression of LLO, PC-PLC, or PI-PLC. PMID- 10826719 TI - Antimicrobial activity of carvacrol toward Bacillus cereus on rice. AB - The antimicrobial activity of carvacrol, a compound present in the essential oil fraction of oreganum and thyme, toward the foodborne pathogen Bacillus cereus on rice was studied. Carvacrol showed a dose-related inhibition of growth of the pathogen. Concentrations of 0.15 mg/g and higher inhibited the growth and the extent of inhibition depended on the initial inoculum size. To decrease the input of carvacrol on the taste and flavor of the product, a combined treatment with the structure analog cymene was tested. Due to the smell and taste of carvacrol at high concentrations, carvacrol was combined with cymene, a natural antimicrobial compound with a similar structure. A synergistic effect was observed when 0.30 mg/g carvacrol was combined with 0.27 mg/g cymene. Finally it was demonstrated that a common taste enhancer like soya sauce also increased the antimicrobial action of carvacrol toward B. cereus. The antimicrobial activity of carvacrol with cymene or soya sauce was influenced by the addition of NaCl. PMID- 10826720 TI - Naturally occurring biofilms on alfalfa and other types of sprouts. AB - Scanning electron microscopy was used to examine the cotyledons, hypocotyls, and roots of alfalfa, broccoli, clover, and sunflower sprouts purchased from retail outlets as well as alfalfa sprouts grown in the laboratory using a tray system equipped with automatic irrigation. Biofilms were observed on all plant parts of the four types of commercially grown sprouts. Biofilms were also commonly observed on alfalfa sprouts grown in the laboratory by 2 days of growth. Rod shaped bacteria of various sizes were predominant on all sprouts examined both as free-living cells and as components of biofilms. Occasionally, cocci-shaped bacteria as well as yeast cells were also present in biofilms. The microbes contained in the biofilms appeared to be attached to each other and to the plant surface by a matrix, most likely composed of bacterial exopolysaccharides. Biofilms were most abundant and of the largest dimensions on cotyledons, sometimes covering close to the entire cotyledon surface (approximately 2 mm in length). Naturally occurring biofilms on sprouts may afford protected colonization sites for human pathogens such as Salmonella and Escherichia coli O157:H7, making their eradication with antimicrobial compounds difficult. PMID- 10826721 TI - Effect of electron beam irradiation on color and microbial bioburden of red paprika. AB - The effect of irradiation with electron beams on the microbiological quality and color properties of red paprika was examined. The irradiation doses ranged from 0 to 12.5 kGy. The counts performed were total mesophilic aerobic microorganisms, Enterobacteriaceae, coliforms, sulfite-reducing clostridia, molds, and yeasts. It was concluded that molds, yeasts, and sulfite-reducing clostridia were the most resistant species, although a 10-kGy dose of irradiation leads to optimum sanitation. Extractable color and apparent color were analyzed to appraise the incidence of the irradiation treatments in the color properties of red paprika. Extractable color was determined according to the American Spice Trade Association method, and apparent color was analyzed by reflectance using the CIELab color space. Data showed no significant differences between the color properties of irradiated and nonirradiated samples. Irradiation was a suitable procedure to minimize the bioburden of red paprika with small modifications of its color properties. PMID- 10826722 TI - Probiotic effects of feeding heat-killed Lactobacillus acidophilus and Lactobacillus casei to Candida albicans-colonized immunodeficient mice. AB - Probiotic bacteria can protect immunodeficient mice from orogastric candidiasis but cause some pathology of their own. Severely immunodeficient patients may be at risk if fed viable probiotics, so this study evaluated the probiotic potential of nonviable probiotic bacteria to protect immunodeficient mice from Candida albicans infections. Heat-killed probiotic bacteria were fed to gnotobiotic bg/bg nu/nu and bg/bg-nu/+ mice to ascertain if they could protect the mice from mucosal and systemic candidiasis. Both heat-killed Lactobacillus acidophilus (HKLA) and heat-killed Lactobacillus casei (HKLC), in comparison to control mice not fed the probiotic bacteria but challenged (oral) with C. albicans, suppressed the severity of orogastric candidiasis in bg/bg-nu/nu mice at 2 weeks after colonization with C. albicans, inhibited disseminated candidiasis in C. albicans colonized bg/bg-nu/+ mice at 4 weeks after colonization, and suppressed the number of viable C. albicans in the alimentary tract. HKLA, but not HKLC, treatment inhibited disseminated candidiasis in bg/bg-nu/nu mice at 2 weeks after oral challenge and enhanced the proliferative responses of splenocytes from C. albicans-colonized bg/bg-nu/+ mice to C. albicans antigens. Neither HKLA nor HKLC were able to prolong the survival of gnotobiotic bg/bg-nu/nu mice after oral challenge with C. albicans. These results demonstrate that heat-killed lactobacilli can induce some (limited) protection (probiotic effect) against candidiasis in mice. PMID- 10826723 TI - Ability of dairy strains of lactic acid bacteria to bind aflatoxin M1 in a food model. AB - Aflatoxin M1 (AFM1) is a highly toxic compound found in milk. Its occurrence poses a threat to the health of consumers, especially young children, and leads to economic losses due to contaminated milk. The problem is global but more severe in developing countries. Consequently, there is a great demand for novel strategies to prevent the contamination and adverse effects of AFM1. To develop a safe and practical decontamination method, a preliminary study was carried out with specific lactic acid bacteria strains that were tested for their ability to remove AFM1 from liquid media. All strains, whether viable or heat-killed, could reduce the AFM1 content of a liquid medium. Two most effective strains were also tested using contaminated skim and full cream milk. The results indicate that specific lactic acid bacteria used in dairy products can offer novel means of decontaminating aflatoxin M1 from milk. PMID- 10826724 TI - A chromogenic medium for the detection of yeasts with beta-galactosidase and beta glucosidase activities from intermediate moisture foods. AB - A selective and differential solid medium for the specific detection of some common yeasts frequently causing spoilage in intermediate moisture foods is described. The principle of the method is based on the detection of two enzymes, beta-glucosidase and beta-galactosidase, using the chromogenic substrates salmon Gluc and X-Gal. Over 140 yeasts and bacteria were tested, and Debaryomyces hansenii and Kluyveromyces marxianus strains produced salmon and dark blue colonies, respectively, thus permitting their clear discrimination from other yeasts common in intermediate moisture foods. The medium was very satisfactory when intermediate moisture foods were tested. PMID- 10826725 TI - Prevalence of Salmonella spp. in poultry broilers and shell eggs in Korea. AB - This study was conducted to determine the presence of Salmonella spp. in raw broilers and shell eggs in Korea. In total, 135 dozen shell eggs and 27 raw broilers were tested. None of the egg yolks were found to contain Salmonella organisms but Escherichia coli, Escherichia hermanii, and Citrobacter freundii were isolated from egg shells. Salmonella spp. were detected in 25.9% of raw broilers, and Salmonella serotypes isolated from raw broilers were Salmonella Enteritidis, Salmonella Virchow, and Salmonella Virginia. D-values and antibiotic resistance of Salmonella isolates were also investigated. D-values of Salmonella enteritidis, Salmonella Virginia, and Salmonella Virchow in tryptic soy broth at 55 degrees C were 2.36, 2.13, and 0.70 min and 0.53, 0.37, and 0.20 min at 60 degrees C, respectively. All Salmonella isolates showed multiple antibiotic resistance patterns and were resistant to penicillin and vancomycin. One strain of Salmonella Enteritidis showed resistance to 12 antibiotics used in this study. PMID- 10826726 TI - Temperature gradient gel electrophoresis of the amplified product of a small 16S rRNA gene fragment for the identification of Listeria species isolated from food. AB - The development of a rapid method for the identification of Listeria spp. is described. It is based on the polymerase chain reaction amplification of a small fragment from the 16S rRNA gene followed by temperature gradient gel electrophoresis. Forty-five strains of Listeria spp. (Listeria monocytogenes, Listeria innocua, Listeria ivanovii, Listeria seeligeri, and Listeria welshimeri) were used for the optimization of the protocol. No differences were observed between the results of the identification of the strains tested using traditional methods and those obtained by polymerase chain reaction-temperature gradient gel electrophoresis analysis. PMID- 10826727 TI - Inactivation of Listeria monocytogenes Scott A on artificially contaminated frankfurters by high-pressure processing. AB - Vacuum-packaged frankfurters, inoculated with 24-h cultures of Listeria monocytogenes Scott A (approximately 10(9) CFU/ml) by injection into the packages, were held at pressures of 300, 500, and 700 MPa for up to 9 min. L. monocytogenes were washed from the surface of the frankfurter and plated onto brain heart infusion agar. During the time to achieve 300, 500, and 700 MPa (come up time), L. monocytogenes populations decreased by 1, >3, and >5 logs, respectively. Additional inactivation of L. monocytogenes occurred while the samples were held at 300 and 500 MPa. A 5-log reduction in bacterial population was possible at all pressure treatments; however, pressurization at 700 MPa showed the fastest inactivation with L. monocytogenes reduced from 10(8) to 10(2) CFU/package during the come-up time. These results show that high-pressure processing may be a viable method for controlling foodborne pathogens in postprocessed, packaged frankfurters. PMID- 10826728 TI - Evaluation of dry sheet medium culture plate (Compactdry TC) method for determining numbers of bacteria in food samples. AB - The Compactdry, a ready-to-use and self-diffusible dry medium sheet culture system, has been developed by the Nissui Pharmaceutical Co. Ltd. for enumerating bacteria in food. The Compactdry consists of special spread sheet with culture medium that is the same as standard method nutrients, a cold water-soluble gelling agent, and a unique plastic dish. The procedure for bacterial examination in a sample solution (1 ml) is to just inoculate a test solution into the center of the self-diffusible medium and incubate at 35 degrees C for 48 h. The Compactdry TC (CTC) for the enumeration of total aerobic bacteria from 97 food samples was compared with the standard plate count (SPC) method and 3M Petrifilm aerobic count plates (PAC). The correlation coefficients between the CTC and SPC method, the CTC and PAC, and the PAC and SPC method were 0.97, 0.99, and 0.97, respectively. The Compactdry system is useful for the enumeration of total aerobic bacteria in food and may be a possible suitable alternative to the conventional pour-plate or the Petrifilm plate methods. PMID- 10826729 TI - Microbiology of potatoes and potato products: a review. AB - Many types of spoilage and pathogenic microorganisms exist on fresh, minimally processed, and fully processed potato products. Potatoes are processed into many products including frozen, dried, ready-to-eat, and minimally processed. The microbiological quality of finished potato products is influenced by the natural microflora, processing, handling, and human contact. The natural microflora of potatoes are influenced by soil and airborne inocula, agricultural practices, harvesting methods, and storage conditions. The microflora of processed products are influenced by all of the factors and conditions affecting the natural microflora as well as the processes applied to the product. Increased consumer demand for new and existing potato products highlights the importance of ensuring their microbiological safety. This review considers the sources of microorganisms, microflora, foodborne disease pathogens, and outbreaks associated with, and selected microbiological research involving, potatoes and potato products. PMID- 10826730 TI - From the shoulders of giants: a look ahead: biotechnology in the 21st century. PMID- 10826731 TI - The case for financial incentives to encourage organ donation. AB - The existing system of organ procurement has failed in its mission of providing an adequate number of organs for transplantation. The organ shortfall results in diminished life expectancy and quality of life for dialysis patients and in increased mortality for patients with end-stage cardiac disease and liver failure who are unable to obtain transplants. Oft-repeated arguments against sale of human organs do not stand up to careful examination, and seem anachronistic in the context of widespread current acceptance of financial incentives for tissue donation in analogous fields, such as reproductive medicine. This essay advocates the staged introduction of a reward-based system of organ donation, initially for cadaveric harvesting and, possibly, later for living donation. Although imperfect, the proposed approach would represent a significant improvement over the status quo. PMID- 10826732 TI - Use of the placenta as an artificial lung. AB - Oxygen transfer across a single cotyledon of the human placenta was assessed by using three different perfusates in the maternal circuit: 1) M-199 culture medium, 2) human adult red blood cells (RBCs), and 3) perflubron. These maternal circuit perfusates were oxygenated with a membrane oxygenator. RBCs were perfused on the fetal side of the circuit and samples were taken preplacenta and postplacenta for each maternal perfusate. PO2 and PCO2 were measured and O2 transfer was calculated for each maternal perfusate. O2 transfer per single cotyledon (mean +/- SE) was 0.18 +/- 0.04, 0.20 +/- 0.03, and 0.15 +/- 0.05 ccO2/min when using: 1) M-199, 2) RBCs, and 3) perflubron, respectively. O2 transfer per kilogram of placental tissue was 13.08 +/- 2.78, 14.57 +/- 2.05, and 10.43 +/- 3.79 ccO2/kg per minute when using: 1) M-199, 2) RBCs, and 3) perflubron, respectively. When extrapolated to the individual weights of the entire placenta, the O2 transfer was 9.15 +/- 1.95, 10.20 +/- 1.43, and 7.30 +/- 2.65 when using: 1) M-199, 2) RBCs, and 3) perflubron, respectively. We conclude from these data that O2 transfer can be accomplished during placental perfusion. Larger studies are required to differentiate efficacy among the three maternal circuit perfusates. PMID- 10826733 TI - Ex vivo testing of the intravenous membrane oxygenator. AB - Intravenous oxygenation represents a potential respiratory support modality for patients with acute respiratory failure or with acute exacerbations of chronic respiratory conditions. Our group has been developing an intravenous oxygenator, the IMO, which uses a constrained fiber bundle and a rapidly pulsating balloon within the fiber bundle. Balloon pulsation drives blood flow past the fibers at greater relative velocities than would otherwise exist within the host vessel, and gas exchange rates are enhanced. The purpose of this study was twofold: (1) to characterize the gas exchange performance of the current IMO in an extracorporeal mock vena cava vessel under conditions of known fixed vessel geometry and controlled blood flow rates; and (2) to compare the IMO gas exchange performance to that reported for the clinically tested IVOX device within a comparable ex vivo set-up. The ex vivo flow loop consisted of a 1 inch ID tube as a mock vena cava that was perfused directly from an anesthetized calf at blood flow rates ranging from 1 to 4 1/2 L/min. O2 and CO2 exchange rates were measured for balloon pulsation rates, which ranged from 0 to 180 bpm. Balloon pulsation significantly increased gas exchange, by 200-300% at the lowest blood flow rate and 50-100% at the highest blood flow rate. Balloon pulsation eliminated much if not all of the dependence of the gas exchange rate on blood flow rate as seen in passive oxygenators. This suggests that in clinical application the IMO may exhibit less gas transfer variability due to differences in cardiac output Over the entire flow rate range studied, the CO2 and O2 gas exchange rates of the IMO at maximal balloon pulsation varied from approximately 250 to 350 ml/min/m2. At maximum balloon pulsation the IMO exchanged CO2 and O2 at rates from 50-500% greater, depending upon the blood flow rate, than the exchange rates reported for the IVOX device in ex vivo tests. PMID- 10826734 TI - Use of reconstructed small intestine submucosa for urinary tract replacement. AB - We used reconstructed SIS (ReSIS), a photocrosslinked biomaterial, to create grafts in various shapes and sizes. Sheets of ReSIS were placed in 14 swine to repair bladder defects, and ReSIS tubes were placed in six swine to replace a segment of excised ureter. Histologic analysis of the bladder repair revealed transitional urothelial cells lining the ReSIS by 1 week. After 2 weeks, fibroblasts and mononuclear cells had infiltrated the ReSIS, neovascularization had occurred, and the urothelial lining was more complex, containing multiple cell layers. After 4 weeks, a definite submucosa was present and the ReSIS was starting to degrade. An initial muscular regeneration was demonstrated at 12 weeks. No foreign body reaction, calcification, or sedimentation was noted in any animal. The ureteral implants showed identical histologic changes, without obstruction or leakage of the replaced segment. The ReSIS allowed rapid urothelial regeneration, ingrowth of new blood vessels, and the orderly deposition and organization of new collagen. Our study demonstrates that the photocrosslinking technique used to create larger sheets and tubes of this biomaterial (ReSIS) does not detract from the positive attributes of the SIS and should improve its usefulness in accomplishing larger bladder augmentations and ureter replacements. PMID- 10826735 TI - Computer simulation of sensorless fuzzy control of a rotary blood pump to assure normal physiology. AB - Rotary blood pumps have been considered effective permanently implantable devices. However, control of such pumps is quite complicated. Sensorless control of pump flow is required because no invasive flow or pressure sensors are wanted. Whereas insufficient pump output can cause underperfusion and should be avoided, overpumping may cause ventricular collapse and must be prevented. An intelligent physiologic control algorithm is highly desirable to reach optimal pump output based on physiologic requirements. We present an intelligent physiologic control mechanism for the blood pump allowing it to achieve normal physiology. Sensorless control of pump flow is gained by analysis of the electric motor current and speed. The required pump output flow is chosen based on heart rate, and an intelligent fuzzy logic based control mechanism is developed to adjust the motor input so that the pump output can reach required flow while also preventing the occurrence of ventricular suction or cannular collapse. Computer simulation was carried out, and the results indicate that the proposed algorithms can achieve required pump flow to obtain normal physiology, whereas overpumping can be prevented to provide safe operation. PMID- 10826736 TI - Access type as a predictor of dialysis adequacy in chronic hemodialysis patients. AB - Dialysis prescription commonly exceeds the delivered dialysis dose. Tunneled hemodialysis catheters (PC) may provide less dialysis than arteriovenous fistula (AVF) and polytetrafluoroethylene grafts (GG), but the impact of access type on the discrepancy (deltaHD) between dialysis prescription and dose is unknown. This study investigates the relationship between deltaHD and vascular access type. Fifty three chronic hemodialysis patients in our unit were prospectively studied for 3 weeks with measurement of delivered single pool and prescribed Kt/V(urea). There were 25 patients with AVF, 17 with GG, and 11 with PC. Demographic characteristics did not significantly differ between groups. Mean prescribed Kt/V(urea) was 1.73 +/- 0.26, and mean delivered Kt/V(urea) was 1.61 +/- 0.26. For 10 of 53 (19%) patients, dialysis delivery was at least equal to that prescribed, and this proportion did not differ between access types. Forty six of fifty three patients (86.7% of all patients) received Kt/V(urea) > 1.3, with no difference in this proportion between access types: AVF 22 of 25 (88.0%), GG 16 of 17 (94.1%), PC 8 of 11 (72.7%). Surprisingly, prescription times for patients with PC (3.6 +/- 0.3 hr) were significantly shorter than for those with AVF (3.9 +/- 0.3 hr) and GG (3.9 +/- 0.3 hr) (p = 0.02), perhaps indicating physician bias toward patients with tunneled catheters. In summary, access type was not a significant predictor of deltaHD, although patients with arteriovenous access tended to receive more dialysis than those with tunneled catheters. While a large proportion of patients received less dialysis than prescribed, the high levels of delivered Kt/V(urea) indicate that adequate dialysis is possible even in patients who must use tunneled catheters. PMID- 10826737 TI - Prediction of equilibrated urea in children on chronic hemodialysis. AB - Urea rebound (UR) after hemodialysis (HD) requires the use of equilibrated urea (Ceq) instead of immediate end-dialysis urea (Ct) for correct quantification of HD, which is impractical. A new formula for predicting Ceq in children is suggested in our study. Thirty eight standard pediatric HD sessions (single pool Kt/V = 1.70 +/- 0.35, K = 4.65 +/- 1.14 ml/min/kg, UF coeff. = 3.2-6.2 ml/h/mm Hg, t = 3.80 +/- 0.46 h) in 15 children (M: 6, F: 9), ages 14.5 +/- 3.28 years were analyzed. Blood samples were taken: before, 70 min from the start, at the end, and 60 min after the end of HD sessions. After correlating UR (20.32 +/- 7.74%) to various HD parameters, we found that it was mainly determined by HD efficiency parameters. Therefore we correlated Ceq to HD efficiency parameters (Ct, urea reduction ratio, Kt/V, and K/V) and found a very high correlation between Ct and Ceq (r = 0.973). Linear regression analysis was used to further investigate this relationship, and a new formula to predict Ceq from Ct was obtained (Ceq = 1.085 Ct + 0.729, R2 = 0.946, SE = 0.49, absolute residuals = 0.38 +/- 0.29 mmol/L). In a validation study (10 HD sessions with new set of urea blood samples) the results obtained by the new formula were compared with measured values of Ceq and those obtained by the Smye formulae. Values predicted by the new formula (9.91 +/- 2.92 mmol/L) were not significantly different from the measured values (10.33 +/- 3.44 mmol/L). Absolute error of the new formula was 0.78 +/- 0.73 mmol/L, median 0.65; ie., 6.93 +/- 5.3%, median 7.7%. Ceq predicted by the Smye formulae (10.95 +/- 4.18 mmol/L) also did not significantly differ from the measured values, but absolute error of predicted values was markedly higher (1.21 +/- 0.90 mmol/L, median 0.89; 11.73 +/- 7.72%, median 10.11%; p < 0.05). When predicted Ceq was used for calculating equilibrated Kt/V (eKt/V), the new formula resulted in lower absolute error (0.09 +/- 0.07, median 0.08) than the Smye method (0.14 +/- 0.08, median 0.12). We conclude that our simple formula is sufficiently accurate in predicting Ceq in standard pediatric HD and that it is more accurate than the existing Smye formulae, while requiring only pre- and post-HD urea samples. We suggest the use of the new formula for predicting Ceq, which can then be used instead of Ct for a more accurate estimation of double pool Kt/V, URR, V, and PCR. PMID- 10826738 TI - Clinical evaluation of a new high efficiency hemodialyzer: polysynthane. AB - Two increasingly common characteristics of the American chronic hemodialysis (HD) population, high hematocrit and large body size, may render the currently recommended adequacy targets difficult to achieve, even with very efficient dialyzers. In a group of patients with these characteristics, we assessed the ability of a new high efficiency dialyzer (PSN210; Baxter Healthcare Corporation) to achieve the currently recommended adequacy targets. Six patients (mean pre-HD weight and hematocrit, 90.3 +/- 18.0 kg and 0.40 +/- 0.03 kg, respectively) were evaluated. At prescribed blood and dialysate flow rates of 400 and 800 ml/min, respectively, and a mean treatment duration of 4 hrs, mean delivered urea Kt/V and reduction ratio (URR) were 1.38 +/- 0.25 and 0.73 +/- 0.07, respectively. For the same flow rates, whole blood clearances for urea, creatinine, and phosphate were 315 +/- 13, 246 +/- 28, and 260 +/- 27 ml/min, respectively. These data indicate this dialyzer has an efficient mass transfer design allowing adequate dialysis to be delivered even to very large patients under high efficiency conditions. PMID- 10826739 TI - In vitro characterization of the occurrence of hemolysis during extracorporeal blood circulation using a mini hemodialyzer. AB - A minimodule dialyzer was made of polyacrylonitrile (PAN) hollow fibers and tested in an in vitro circuit under varying flow rates and blood volumes. Hemolysis occurred in the open circuit in the inlet and outlet chambers. The module was measured to investigate the hemolysis effect during dialysis. The results show that hemolysis ratio (HR) increased with blood flow rate. When the blood speed was increased from 27 to 70 cm/sec, the damaged fraction of red blood cells increased from 0.8% to 1.5%. Shearing in the hollow fiber contributed approximately 13% of the overall HR. Approximately 55% of the overall HR occurred at the inlet and outlet chambers. With increasing blood volume, the contacting surface area per unit blood volume (S/V) and the frequency of pump squeezing of red cells were decreased; hence the damage to the red blood cells and thus reduction of HR. When S/V was 2 cm(-1), the damaged fraction of red cells was approximately 0.7%. In addition, the tubing and connectors of the circuit caused further damage to the red blood cells. By referring to the linear velocity and blood volume, we can predict the HR in an actual protocol from the results of this experiment. This should improve the quality of hemodialysis and benefit the patient. PMID- 10826740 TI - Analysis of the long-term efficacy and selectivity of immunoadsorption columns for low density lipoprotein apheresis. AB - Immunoadsorption low density lipoprotein (LDL) apheresis is performed with reusable columns containing anti-apolipoprotein B(ApoB) antibodies. We analyzed their long-term efficacy and selectivity. Performance over 60 treatment sessions of six pairs of immunoadsorption LDL apheresis columns was evaluated by analysis of variance using the removal of total cholesterol and ApoB to assess efficacy and the ratio of total cholesterol/high density cholesterol removed to assess selectivity. The removal of cholesterol did not vary significantly with treatment number. The mass of ApoB removed increased significantly (p = 0.002), and the mass of ApoB removed per volume unit of processed plasma showed a trend (p = 0.065) toward an increase with treatment number. Both parameters correlated with the serum ApoB concentration before treatment, which also increased significantly (p = 0.0007) with treatment number. No significant variation of selectivity was found. The efficacy of the LDL apheresis immunoadsorption columns did not decrease after 60 treatment sessions. The columns' selectivity also remained unchanged. PMID- 10826741 TI - Thromboembolization associated with sudden increases in flow in a coronary stent ex vivo shunt model. AB - To observe the dynamics of thromboembolism (TE) in an animal model, a carotid carotid arterial ex vivo shunt was developed. A coronary stent deployed in a 3.5 mm polyvinylchloride (PVC) tubing segment was used as a model device in the shunt. A light-scattering microemboli detector monitored the embolic content of the blood flowing through the shunt at 50-150 ml/min as determined by a clamp-on ultrasound flow probe. The stent was found to actively develop thrombi and release emboli for 1-3 hours when the activated clotting time (ACT) was maintained between 125 and 150 seconds. The shunt flow rate fluctuated considerably (from 50 to 150 ml/min) depending on the animal's activity. When the time profile of this fluctuating flow rate was super imposed on the time profile of embolization, it was noted that sudden increases in flow rate were associated with incidents of embolization. Statistically, sudden flow rate increases of 100% or more were accompanied by embolic events 95% of the time (p < 0.01). Based on the results of this study, it was postulated that the increased embolization may be due to the fluid forces associated with accelerating flow. To explore this postulate, in vitro studies were conducted to compare the effects of pulsatile flow with steady flow on stent induced TE. Results of this study suggested a significant increase (100%) in both stent thrombosis and embolism during pulsatile flow compared with steady flow. PMID- 10826742 TI - Angiogenesis in the latissimus dorsi muscle using different regimens of electrical stimulation and pharmaceutical support. AB - It is our contention that the prevention of ischemia-reperfusion injuries immediately after latissimus dorsi muscle (LDM) mobilization and enhancement of angiogenesis will be effective in improving cardiomyoplasty results. The investigations were performed on adult sheep. Three hours after LDM mobilization, various stages of leukocyte-endothelium interaction were revealed: leukocytes binding to the endothelium, leukocyte destruction of endothelium, and leukocytes leaving capillaries through gaps in the endothelium. Fifty-six days after mobilization various stages of necrosis were discernible. The area occupied by capillaries was 3.45 +/- 0.26% vs. 3.99 +/- 0.24% in control muscle; most of the endothelial cells exhibited morphologic degeneration. Electrical stimulation with 60 CPM actually decreased the capillary density to 2.15 +/- 0.7%, and most of the endothelial cells were damaged, with disrupted plasma membranes. Muscle subjected to 15 CPM increased the percent of capillaries to 5.01 +/- 0.56%, and endothelial cells appeared normal in ultrastructure. Pharmaceutical support prevented muscle damage and accelerated revascularization. After 56 days of autologous biological glue (ABG) application, the area occupied by capillaries was 5.57 +/- 0.24%. This increased to 8.47 +/- 0.72% when aprotinin (proteinase inhibitor) was added to ABG, and to 9.40 +/- 1.24% with pyrrolostatin (free radical scavenger). Both ABG application with aprotinin and electrical stimulation at 15 CPM prevent the LDM from postmobilization damage, and increase angiogenic potential. PMID- 10826743 TI - Effect of heparin reversal and fresh platelet transfusion on platelet emboli post cardiopulmonary bypass in a pig model. AB - Heparin reversal by protamine and fresh platelet transfusion may decrease bleeding complications post-cardiopulmonary bypass (CPB) and may increase the level of organ trapped platelet emboli. Platelet emboli were quantified in two groups of 12 Yorkshire pigs (30-35 kg), where 111indium labeled autologous platelets (INPLT: 850-1,200 microCi) were injected intravenously before and after CPB (BCPB, ACPB), and the platelet emboli level in intact organs and their samples (brain, heart, kidneys, lung, liver, and spleen) was quantified with an ion chamber and a gamma counter, respectively. All pigs were systemically heparinized (ACT > 400 sec). CPB was carried out at 2.5-3.5 L/min at 28 degrees C using a centrifugal pump, an oxygenator (OX:Bentley Univox 1.8 m2), an arterial filter (AF:0.25 m2), and a cardiotomy reservoir (CR: BMR 250) for 90 min. Heparin was reversed with an equivalent dose of protamine. The percent of INPLT dose (ID%, mean +/- SD) in organs of BCPB and ACPB pigs was calculated. The sequence of platelet emboli on a unit weight basis (ID%/g) had the following order: Spleen > Liver > Lung > Kidneys > Heart > Brain. The presence of significantly higher levels of emboli in brain, heart, and kidneys in the ACPB than the BCPB group suggest that platelet transfusion after heparin reversal with protamine may increase the risk of platelet emboli. However, it is an acceptable risk for patients having bleeding complications post-CPB. PMID- 10826744 TI - Prevalence and persistence of heparin/platelet factor 4 antibodies in patients with heparin coated and noncoated ventricular assist devices. AB - Thromboembolism is a major complication in patients with ventricular assist devices (VAD). Anticoagulation with heparin, coumarin, and anti-platelet agents, particularly the development of biocompatible surfaces such as inner pseudo endothelial layers or a coating with heparin, are intended to reduce these complications. However, the administration of heparin can lead to heparin induced thrombocytopenia type II (HIT II). Predominantly heparin/platelet factor 4 (HPF4) antibodies are responsible for the development of HIT II. The goal of the present investigation was to assess the prevalence of these antibodies in patients with heparin coated and noncoated VADs. Fifty-five patients were enrolled in the investigation. A heparin coated system was implanted in 30 patients, and a noncoated system was implanted in 25 patients. Antibodies were evaluated before, on days 7 and 14, and 3 months after implantation. Testing was performed with the Heparin/Platelet factor 4 enzyme-linked immunosorbent assay (ELISA) (Stago, France). In 40 of the 55 patients, the formation of HPF4 antibodies was observed (73%). In 35 of these patients (88%), HPF4 antibodies were present before surgery. There were no differences between the groups. In 11 patients (equal from both groups), the antibodies disappeared after termination of systemic heparinization. We conclude that in a rather high percentage of patients with VADs HPF4 antibodies are found. This finding may be explained by the repetitive and prolonged exposure of these patients to heparin. Immobilized heparin, as presently used in the carmeda coating, seems not to influence the formation and persistence of HPF4 antibodies. Further studies will have to prove whether HPF4 antibodies contribute to thromboembolic complications in these patients. PMID- 10826745 TI - Hydrodynamic properties of a new percutaneous intra-aortic axial flow pump. AB - Cardiac intervention, myocardial infarction, or postoperative heart failure will sometimes create a need for circulatory support. For this purpose, a new, minimally invasive intra-aortic cardiac support system with a foldable propeller has been developed. In animals, the pump has been shown to have a positive hemodynamic influence, and the present study evaluates the hydraulic properties of the pump in a bench test. The axial flow pump is a catheter system with a distal motor driven foldable propeller (0-15,000 revolutions per minute). To protect the aortic wall, filaments forming a cage surround the propeller. In the present study, tests were done with two different pumps, one with and one without the cage. Two different models were used, one for testing pressure generation and one for obtaining flow-pressure characteristics. Propellers and tubes with different diameters were studied, and pressure and flow characteristics were measured. The mathematical relationships between pressure and rotational speed, pressure, and diameter of propeller and tube were determined. There was a positive relationship between the revolutions per minute and the generated pressure, a positive relationship between the diameter of the propeller and pressure, and a negative relationship between the diameter of the tube and the generated pressure. Within the physiologic range of cardiac output, there was a small drop in pressure with increasing flow in the tubes with a small diameter. With an increasing diameter of the tube, a smaller pressure drop was seen with increasing flow. The present cardiac support system has hydraulic properties, which may be of clinical relevance for patients with left ventricular heart failure. PMID- 10826746 TI - Outcome of patients with ventricular assist devices and acute renal failure requiring renal replacement therapy. AB - The significance of acute renal failure (ARF) for patients treated with a ventricular assist device (VAD) is uncertain. There is little information on the outcome of patients who require renal replacement therapy during treatment with a VAD. A retrospective review was undertaken to evaluate the impact of renal failure requiring renal replacement therapy on such patients. Studied were 227 patients who were supplied with a VAD at the German Heart Institute Berlin. Fifty five patients required renal replacement therapy during treatment with a VAD. These were compared with patients not needing renal replacement therapy (ARF and non-ARF groups). Significant differences for the end points of survival, heart transplantation, and discharge from hospital were observed in patients with ARF (p < 0.01). Survival was then analyzed according to indications for treatment with a VAD (bridge to transplantation or cardiac recovery after cardiotomy, transplantation, myocardial infarction, myocarditis, and endocarditis). Survival for bridge-to-transplantation patients was clearly influenced in a negative way by ARF (p < 0.01). For cardiac recovery patients, only a small difference in survival was observed (p = 0.05). We conclude that ARF is a negative predictor for bridge-to-transplantation patients. For cardiac recovery patients the impact of ARF on survival is marginally significant. PMID- 10826747 TI - P-selectin monoclonal antibody may attenuate the whole body inflammatory response induced by cardiopulmonary bypass. AB - Cardiopulmonary bypass (CPB) is known to induce an inflammatory response in association with neutrophil mediated lung injury. P-Selectin has been reported to be involved in the initiation of this inflammatory response by promoting the adhesion of neutrophils to endothelial cells in postcapillary venules. However, the role of P-selectin in the inflammatory response induced by CPB has never been clarified. To elucidate its role, we evaluated the effect of an anti-rat specific P-selectin monoclonal antibody (ARP2-4; Sumitomo Pharmaceutical) on the response of inflammatory cytokines and lung injury in a rat-CPB model. Twenty Sprague Dawley rats underwent CPB for 30 minutes (80 ml/kg per minute, 34 degrees C) under one of two conditions. In group P, ARP2-4 (3 mg/kg) was added to the priming solution of the bypass circuit (n = 10). Saline alone was given to group C (n = 10). Inflammatory cytokines (tumor necrosis factor-alpha [TNF-alpha], interleukin[IL]-1beta, IL-6, and IL-8) and respiratory index (RI) as a marker of pulmonary gas-exchange ability were measured 1) before the initiation of CPB, 2) at the termination of CPB, and 3) 2 hours after the termination of CPB. Neither TNF-alpha nor IL-1beta was detected during the experimental period in either group. The plasma levels of IL-6 and IL-8 increased after CPB in both groups, but they were significantly lower in group P than in group C. The RI value increased in a pattern similar to that of the inflammatory cytokines and was significantly lower in group P. These data demonstrate that the addition of an anti-rat specific monoclonal antibody inhibits the abnormal release of inflammatory cytokines and attenuates CPB induced lung injury in rats. Thus, P-selectin may play a role in the augmentation of CPB induced inflammatory response, and the use of its inhibitory monoclonal antibody seems to be a promising strategy for the treatment of CPB induced lung injury. PMID- 10826748 TI - Development of an implantable small right ventricular assist device. AB - Currently, at least two permanent implantable left ventricular assist devices (LVADs) are used clinically. Unfortunately, there is no small implantable right ventricular assist device (RVAD) available, even though at least 25-30% of this patient population has right ventricular failure. If a small implantable RVAD were available, biventricular assist could support patients with right ventricular failure. A small atraumatic and antithrombogenic RVAD is being developed to meet this clinical need. This small centrifugal blood pump, the Gyro PI pump, is 6.5 cm in diameter and 4.6 cm in height and has three unique characteristics to prevent thrombus formation: (1) the double pivot bearing and magnetic coupling system enable this pump to be completely sealless; (2) the secondary vanes at the bottom of the impeller accelerate the blood flow and prevent blood stagnation; and (3) the eccentric inlet port enables the top female bearing to be embedded into the top housing and decrease blood cell trauma. The inflow conduit consists of a wire reinforced tube and a hat-shaped tip that is biolized with gelatin to create a thrombus resistant material. This conduit is directly implanted into the right ventricle, and the outflow conduit is anastomosed to the PA. The pump can be implanted inside the abdominal wall or in the thoracic cavity. Biocompatibility of this pump was proved in two calves by thrombus free implantation as an LVAD for 284 days and 200 days. Two RVAD implantations were conducted, aiming for 1-month system feasibility studies. During the month, the RVADs operated satisfactorily without any thromboembolic incident. No blood clots or abnormal findings were seen inside the pump, nor were there abnormal findings in the explanted lungs except for small areas of atelectasis. The pump flow was 3.02 +/- 0.38 L/min in calf 1 and 3.75 +/- 1.18 L/min in calf 2. The power requirement was 7.28 +/- 0.43W for calf 1 and 14.52 +/ 3.93W for calf 2. The PaO2 was 72.0 +/- 3.60 mm Hg (calf 1) and 72.0 +/- 7.63 mm Hg (calf 2); PaCO2 was 38.3 +/- 2.17 mm Hg (calf 1) and 34.1 +/- 1.95 mm Hg (calf 2); and SaO2 was 94.1 +/- 1.37% (calf1) and 95.0 +/- 1.95% (calf 2). Gas exchange via the lungs was maintained. These studies indicate that the Gyro PI pump is suitable as a single implantable RVAD, and is a feasible RVAD as a part of a BiVAD system in terms of pump performance and thrombus resistance. PMID- 10826749 TI - A bovine model for detecting high intensity transient signals originating from mechanical heart valves. AB - In patients with mechanical heart valves (MHVs), transcranial Doppler methods commonly detect high intensity transient signals (HITS) representing microemboli. These microemboli, which are presumably gaseous, may cause stroke and cognitive deterioration. A bovine model was therefore developed for studying the relationship between mitral MHV induced HITS and potential etiogenic factors. We placed an 18 mm, 4 MHz Doppler probe in the brachiocephalic artery to detect MHV induced microbubbles at baseline (rest) and under 9 other conditions. To elucidate the gas composition (CO2 or N2) of the microbubbles, we administered 1%, 3%, and 5% CO2, and 100% O2. To determine effect of the heart rate, we paced the heart at 120, 160, and 180 bpm. To alter the myocardial contractility, we gave dobutamine and esmolol. Two independent, blinded observers counted the HITS from recorded doppler spectra. HITS were defined by an initial unidirectional spectral deviation, a signal power of >8 dB relative to the background power, and lack of a cyclic appearance. The electrocardiogram, aortic and LV pressures, and LV dP/dt were obtained telemetrically. The calves were studied 4 to 6, 8 to 10, and 12 to 14 weeks postoperatively, after which the animals were sacrificed at an approximate 4 month study duration, and a postmortem evaluation of the heart and the main viscera was performed. In all, 27 HITS recordings were made in 10 calves. Myocardial contractility was the only factor to significantly affect HITS frequency; the heart rate and blood gas concentrations had minimal effect on HITS frequency. Our bovine model will be useful for assessing valve designs, as well as the mechanism of HITS, the composition of the microemboli, and their possible pathophysiologic effects on the kidneys and brain. PMID- 10826750 TI - Development of an intracorporeal Thoratec ventricular assist device for univentricular or biventricular support. AB - There is a need for a small, simple, and versatile intracorporeal ventricular assist device (IVAD) as an alternative to the large implantable electromechanical LVAD systems in current use. Because the basic design of the Thoratec paracorporeal VAD has been demonstrated in over 1,000 patients, weighing from 17 to 144 kg, and for durations up to 515 days including patient discharge (by using the portable driver), we are developing a new intracorporeal version of our VAD. This IVAD has a smooth contoured, polished titanium housing, and maintains the same blood flow path and Thoralon polyurethane blood pumping sac as the paracorporeal VAD. The IVAD is controlled with the Thoratec TLC-II Portable VAD Driver, which is a small briefcase sized, battery powered, pneumatic control unit. Intracorporeal LVADs and/or RVADs are implanted in a preperitoneal position, with a single small (9 mm OD) percutaneous pneumatic driveline for each VAD. The major advantages of the new IVAD design are size and simplicity. The IVAD weight (339 g) and implanted volume (252 ml) are substantially smaller than current implantable electromechanical LVAD systems. Only the small blood pump is implanted, leaving the more complex control unit external, where it can be serviced and replaced. The versatile design is intended for left and/or right heart support in large or small patients. The IVAD in combination with the TLC-II portable driver will be a viable and attractive alternative to large, implanted electromechanical systems. PMID- 10826751 TI - Anticoagulation during use of a left ventricular assist device. AB - Thirty-six mongrel dogs underwent 24hr left ventricular assist. The VAD was placed between the left atrium and the descending aorta, and the dogs were divided into four groups according to type of anticoagulation: no anticoagulation, argatroban, nafamostat mesylate, and nafamostat mesylate + prostacyclin analog. Results of this animal experiment revealed that a newly developed synthetic thrombin inhibitor argatroban can prevent activation of the intrinsic coagulation pathway. Argatroban is efficient under any blood coagulative condition, even lack of anti-thrombin III, because of its direct inhibitory effect on thrombin, making argatroban more useful than heparin as an anticoagulant for LVAD. Argatroban, as well as heparin, provides marked and significant prolongation of the prothrombin time from early assisted circulation, but produces a bleeding tendency. Nafamostat mesylate can maintain blood coagulation parameters within the acceptable range. Combined administration of nafamostat mesylate and a prostacyclin analog cause the least decrease in fibrinogen and alpha2-plasmin inhibitor among the four groups and causes no significant prolongation of prothrombin time. PMID- 10826752 TI - Pump perfusion causes vasodilation by activation of platelets. AB - Use of a pump in extracorporeal circuits depresses autoregulation and vascular tone. To study whether platelets are involved, we perfused rat hindlegs by means of an extracorporeal shunt between carotid and femoral artery. Autoperfusion could instantaneously be replaced by pump perfusion. To avoid interference by effects caused by blood-material contact, the circuit was coated with albumin. Spontaneous flow did not elicit platelet aggregation as recorded continuously with a photometric device inserted into the tubing, nor did it affect femoral vascular resistance. However, pump perfusion immediately evoked strong platelet aggregation that stabilized at a lower level after 2-3 minutes. Femoral resistance rose slightly during the first 2 minutes, but thereafter fell to 63% of control and stayed at approximately 70% for the next 2 hours. Pump induced platelet aggregation and fall in vascular resistance could be prevented with aurintricarboxylic acid, which specifically inhibits shear induced platelet aggregation. We conclude that pump perfusion with blood in coated systems elicits shear-induced platelet aggregation that, in turn, leads to vasodilation in the perfused vascular bed. These effects can be prevented by blocking the binding of von Willebrand factor to the platelet glycoprotein Ib receptors. PMID- 10826753 TI - Recovery directed left ventricular assist device: a new concept. AB - To promote cardiac recovery, we developed a recovery directed left ventricular assist device (RDLVAD) that consists of a valved apical conduit, an afterload controlling chamber (ACC), and a pump. We evaluated its efficacy by comparison with an ordinary LVAD. In each of six pigs with ischemia-induced heart failure, flow and pressure measurements were made while maintaining the total blood flow and arterial pressure equal in the two groups. RDLVAD was able to direct all the blood ejected from the LV into the ACC (0-15 mm Hg) but not into the aorta (73 mm Hg). In the ordinary LVAD, however, some ejection occurred into the aorta despite vigorous suction of the LV. Thus, RDLVAD increased DPTI/SPTI 2.3 times (p < 0.005) and decreased left ventricular end-diastolic pressure by 40% and maximum dP/dt by 20% (p < 0.05). Even the apical valve, at approximately half the diameter of the aortic valve, was able to allow all the blood ejected from the LV to enter the ACC. In one control group pig that achieved almost no ejection into the aorta, left ventricular relaxation and dilatation was extremely limited. RDLVAD may promote cardiac recovery by ensuring less LV work, a greater blood supply/demand ratio in the coronary circulation, and full ventricular relaxation. PMID- 10826754 TI - Human factors issues in ventricular assist device recipients and their family caregivers. AB - Ultimately, for ventricular assist devices (VADs) to be acceptable as permanent alternatives to heart transplantation, patients' and their families' satisfaction with specific features and risks of VADs must be addressed. Of 42 eligible patients who received VADs between February of 1996 and December of 1998, we interviewed 37 patients (17 Novacor, 18 Thoratec, 2 with both devices) and 20 of their primary family caregivers about device related concerns and reactions. Demographic and health related correlates of respondents' concerns were examined. Eleven patients discharged from the hospital with the VAD in place were then reinterviewed 1 month after discharge. At baseline, patients' general perceptions of the VAD were positive, although 22-52% reported specific concerns, including most often worry about infection (52%), difficulty sleeping due to the position of the driveline (52%), pain at the driveline exit site (46%), worry about device malfunction (40%), and being bothered during the day by device noise (32%). The prevalence of most concerns rose with duration of VAD support. Caregivers' perceptions did not differ significantly from patients' perceptions. Outpatients were somewhat more concerned than inpatients about device noise and risk of stroke, but were markedly less concerned about infection. Across all patients, higher levels of device related concerns were correlated with more physical functional limitations and more psychological distress, and reduced quality of life. Demographic characteristics and device type were not uniformly related to device concerns. PMID- 10826755 TI - The oculoplastic surgeon as gardener. PMID- 10826756 TI - Sentinel lymph node mapping for patients with cutaneous and conjunctival malignant melanoma. PMID- 10826757 TI - Clinical features predictive of orbital exenteration for conjunctival melanoma. AB - PURPOSE: Conjunctival malignant melanoma can sometimes be difficult to control locally, and orbital exenteration may be necessary. To our knowledge, the risk factors that portend exenteration have not been previously identified. The purpose of this study was to review patients who underwent orbital exenteration for advanced disease and to identify retrospectively the clinical factors predictive of ultimate exenteration. METHODS: The records of patients with conjunctival melanoma who were managed on the Oncology Service at Wills Eye Hospital from 1974 to 1997 were reviewed. Factors predictive of orbital exenteration were analyzed using Cox proportional hazards regression models. RESULTS: Of 151 consecutive patients with conjunctival melanoma, 20 (13%) required orbital exenteration for advanced disease. The number of melanoma excisions performed before referral ranged from 0 to 11, with a mean of 2. The clinical factors predictive of orbital exenteration on multivariable analysis were visual acuity of 20/200 or worse, lack of tumor pigmentation, and extralimbal tumor location. All exenterations were performed using an eyelid sparing technique. At mean follow-up of 51 months after exenteration, 4 patients (20%) had died of metastasis, 3 (15%) were alive with metastasis, and 13 (65%) had not developed metastasis. CONCLUSIONS: Conjunctival melanoma can show aggressive local behavior, and orbital exenteration is necessary in up to 13% of cases at a tertiary referral center. Patients with risk factors such as poor visual acuity, amelanotic tumor, and extralimbal tumor location may require wider excision and radiation therapy to control the tumor and avoid orbital exenteration. It is hoped that modern surgical approaches will decrease the need for orbital exenteration. PMID- 10826758 TI - Correlation of preoperative computed tomography and postoperative ocular motility in orbital blowout fractures. AB - PURPOSE: To determine a relationship between preoperative soft tissue disruption and postoperative ocular motility in orbital blowout fractures. METHODS: This retrospective cohort study reviewed 30 patients who met all criteria: retrievable coronal computed tomography (CT) scans; internal fractures of the orbital floor, with or without medial wall extension; preoperative diplopia; repair by a single surgeon; complete release of entrapped tissues; and postoperative binocular visual fields (BVFs). Motility outcomes were quantified by one group of the authors, who measured the vertical fusion within BVFs. Other authors analyzed CT scans, designating each fracture as either A or B, based on lesser or greater soft tissue distortion relative to the configuration of bone fragments. The interval between trauma and surgery was also determined. RESULTS: Among the 15 patients with a postoperative motility outcome poorer than the median (86 degrees or less), four (27%) had A fractures; 11 (73%) had B fractures. Among the 15 patients with an outcome better than the median (88 degrees or more), 10 (67%) had A fractures; five (33%) had B fractures. Differences were more defined away from the median. Among five patients with B fractures and better than the median result, three (60%) had surgical repair during the first week after injury. Among the 11 patients with B fractures and less than the median result, one (9%) had repair during the first week. CONCLUSIONS: Postoperative motility is influenced by soft tissue-bone fragment relationships. Whether the outcome can be altered by earlier surgery in selected cases will be determined by prospective studies. PMID- 10826759 TI - Mechanisms of orbital floor fractures: a clinical, experimental, and theoretical study. AB - PURPOSE: The purpose of this study was to investigate the two accepted mechanisms of the orbital blowout fracture (the hydraulic and the buckling theories) from a clinical, experimental, and theoretical standpoint. METHODS: Clinical cases in which blowout fractures resulted from both a pure hydraulic mechanism and a pure buckling mechanism are presented. Twenty-one intact orbital floors were obtained from human cadavers. A metal rod was dropped, experimentally, onto each specimen until a fracture was produced, and the energy required in each instance was calculated. A biomathematical model of the human bony orbit, depicted as a thin walled truncated conical shell, was devised. Two previously published (by the National Aeronautics and Space Administration) theoretical structural engineering formulas for the fracture of thin-walled truncated conical shells were used to predict the energy required to fracture the bone of the orbital floor via the hydraulic and buckling mechanisms. RESULTS: Experimentally, the mean energy required to fracture the bone of the human cadaver orbital floor directly was 78 millijoules (mJ) (range, 29-127 mJ). Using the engineering formula for the hydraulic theory, the predicted theoretical energy is 71 mJ (range, 38-120 mJ); for the buckling theory, the predicted theoretical energy is 68 mJ (range, 40-106 mJ). CONCLUSION: Through this study, we have experimentally determined the amount of energy required to fracture the bone of the human orbital floor directly and have provided support for each mechanism of the orbital blowout fracture from a clinical and theoretical basis. PMID- 10826760 TI - Comparison of oxymetazoline and lidocaine versus cocaine for outpatient dacryocystorhinostomy. AB - PURPOSE: To evaluate the effectiveness of oxymetazoline combined with lidocaine versus cocaine for outpatient dacryocystorhinostomy (DCR). METHODS: Twenty-seven patients underwent DCR with a combination of oxymetazoline 0.05% nasal spray and lidocaine 4% spray/pledgetts (oxy/lido) and were compared with 28 patients who underwent DCR with cocaine 4% pledgetts for nasal anesthesia and vasoconstriction. Most of the patients underwent standard external approach DCR: five patients in the oxy/lido group and four patients in the cocaine group underwent endoscopic DCR. RESULTS: Patients in both groups reported being comfortable during and immediately after surgery. Visualization of the operative field was adequate, and surgery was successfully completed in all cases. Mean blood loss was 6.3 ml (+/-6.0) in the oxy/lido group and 7.3 ml (+/-6.4) in the cocaine group (p = 0.541). There were no cases of postoperative epistaxis requiring nasal packing or hospital admission in the oxy/lido group, whereas one patient in the cocaine group developed epistaxis 8 days after surgery, which required treatment. Success rate was similar in the two groups. CONCLUSIONS: The combination of oxymetazoline and lidocaine is effective for outpatient dacryocystorhinostomy. Patient comfort was adequate and blood loss not significantly different in comparison to cocaine. This combination provides effective nasal anesthesia and vasoconstriction, while avoiding the "hassle" and potentially greater risks associated with cocaine. PMID- 10826761 TI - Sebaceous cell carcinoma of the eyelid and the human immunodeficiency virus. AB - PURPOSE: Patients infected with the human immunodeficiency virus (HIV) are at increased risk for developing malignancies, which are acquired at a younger age and are more aggressive. Sebaceous cell carcinoma is a rare eyelid tumor typically occurring in the seventh decade of life. We report two cases of sebaceous cell carcinoma in young HIV-infected patients. METHODS: Case series and review of the literature. We describe two HIV-infected patients with sebaceous cell carcinoma of the eyelid and caruncle. The first patient was a 36-year-old woman with a 9-month history of an enlarging right lower eyelid mass. The second patient was a 34-year-old man with a 6-month history of an enlarging right caruncular mass. RESULTS: Biopsy showed both masses to be sebaceous cell carcinoma. The first patient underwent Mohs' micrographic excision of the lesion followed by reconstruction of the full-thickness eyelid defect with a combination of tarsoconjunctival and myocutaneous advancement flaps. The second patient underwent exenteration because of orbital involvement. CONCLUSION: Sebaceous cell carcinoma should be considered for any suspicious eyelid lesion in young HIV infected patients. PMID- 10826762 TI - Radiation therapy for local control of eyelid sebaceous cell carcinoma: report of two cases and review of the literature. AB - PURPOSE: Because of previous reports of increased rate of recurrence and mortality after radiation therapy, eyelid sebaceous cell carcinoma has been considered radioresistant. Recent reports of success with primary radiation therapy have been attributed to advancements in irradiating technology and technique. Two cases of eyelid sebaceous cell carcinoma successfully treated with radiation therapy are reported. The techniques used are compared with the techniques described in previous reports, and factors favoring successful treatment are reevaluated. METHODS: Case series and review of the literature. Two cases of eyelid sebaceous cell carcinoma that underwent radiation therapy are described. Both patients were offered, but refused, surgical excision. One patient received 69 Gy combined superficial and megavoltage x-ray irradiation to the left lower eyelid. The second patient received 59 Gy megavoltage electron beam irradiation to the right upper eyelid. A review of the literature was performed, and Fisher's exact test analysis was used to compare the results of all reported cases treated with < or =55 Gy with those treated with >55 Gy. RESULTS: In both cases, the tumor responded to radiation therapy. One patient died 39 months after treatment, of myocardial infarction. The second patient is without clinical evidence of tumor recurrence 46 months after treatment. Fisher's exact test showed an advantage to patients treated with >55 Gy radiation (p = 0.05). CONCLUSION: Radiation therapy with an appropriate delivery system is effective as a curative treatment for eyelid sebaceous cell carcinoma when >55 Gy of radiation dose is delivered. It should be considered for patients seeking an alternative to surgical excision. PMID- 10826763 TI - Chronic exposure of hydroxyapatite orbital implants: cilia implantation and epithelial downgrowth. AB - PURPOSE: To describe previously unreported histologic findings in two patients who developed chronic implant exposure and abscess formation within hydroxyapatite orbital implants. METHODS: Surgically removed implant specimens were processed for histopathologic examination and stained for microorganisms. Each patient's clinical course, socket appearance, and exposure management were reviewed. RESULTS: Histopathologic examination of case 1 showed a channel of necrosis leading from the anterior surface of the implant to its center. Hair shafts were discovered embedded within this channel. Histopathologic examination of the site of chronic exposure in case 2 showed epithelial ingrowth into the pores of the implant. Both spheres in this report indicated limited fibrovascular ingrowth and abscess formation. CONCLUSIONS: Chronic exposure of hydroxyapatite implants allows a portal of entry for extraneous hair shafts and also can lead to epithelial downgrowth. Both of these may be contributing factors in the development of serious implant infections. PMID- 10826764 TI - The safety and applications of acellular human dermal allograft in ophthalmic plastic and reconstructive surgery: a preliminary report. AB - PURPOSE: To describe the uses and determine the safety of a commercially available acellular dermal allograft in ophthalmic plastic and reconstructive surgery. The existing literature regarding current applications and results using acellular dermal allograft is reviewed. METHODS: The study design was a retrospective, noncomparative case series. Participants consisted of 63 consecutive patients undergoing ophthalmic plastic and reconstructive surgery using an acellular dermal matrix graft. The main intervention was placement of a dermal allograft. Outcome measures included the degree of clinical improvement and complications for each patient. RESULTS: We used AlloDerm (LifeCell Corporation, The Woodlands, TX, U.S.A.) as a posterior lamellar conjunctival spacer graft, a soft tissue interpositional graft, and an orbital implant wrapping material. Clinical improvement was noted in all cases. There were no complications attributable to the acellular dermal matrix material. CONCLUSIONS: We describe several novel ophthalmic plastic and reconstructive surgical applications using acellular dermal allograft tissue. This material may be used safely as a posterior lamellar conjunctival spacer graft, a soft tissue interpositional graft, and a wrapping material. Clinically, the allograft appears to be biocompatible, nontoxic, and nonallergenic in the orbit, eyelid, and midfacial tissues. Long-term data and studies comparing the efficacy of acellular dermal allograft with conventional materials are necessary. PMID- 10826765 TI - Outpatient ophthalmic plastic surgery: outcomes and patient satisfaction using initial postoperative telephone call follow-up. AB - PURPOSE: To evaluate the safety and efficacy of using a telephone call within 24 hours as the initial follow-up after outpatient ophthalmic plastic surgery, deferring the first postoperative visit 4 to 7 days. METHODS: We prospectively evaluated clinical outcomes, including complications and patient satisfaction, after 469 outpatient eyelid, lacrimal and anterior orbital procedures. Patient satisfaction was assessed with a 13-item questionnaire. RESULTS: Response to the questionnaire was 282 of 469 (60%) patients. Essentially, all patients responding to the questionnaire who received a telephone call believed that it was helpful and their questions and concerned were adequately addressed. Only 4 of 274 (1%) patients without a planned 24-hour visit indicated that they would have preferred being seen in the office. Complications of a minor nature were noted in 12 of 469 (3%) patients, none of which adversely affected clinical outcomes. CONCLUSION: We conclude that a telephone call on the initial postoperative day, deferring the first visit 4 to 7 days, is a safe, cost-effective strategy with high patient acceptance for most types of eyelid and lacrimal surgery, as well as some types of orbital surgery. PMID- 10826766 TI - Central corneal dellen: a complication of upper eyelid tarsotomy. AB - PURPOSE: To report a rare complication of central corneal dellen that developed after a tarsotomy procedure to correct cicatricial entropion. METHOD: Central corneal dellen developed in a 60-year-old woman 3 months after a tarsotomy procedure for correction of cicatricial upper eyelid entropion. Eversion of the upper eyelid showed a retracted and buckled tarsal plate and the presence of a 5 x 10-mm depression between the edge of the tarsoconjunctival flap and lower tarsal bridge. RESULTS: The recalcitrant corneal defect was reversed when the upper eyelid tarsoconjunctival surface contour was restored by full-thickness tarsal incisions designed to flatten the dome-shaped tarsal plate. CONCLUSION: An upper eyelid defect producing suboptimal lid-globe apposition during lid closure can lead to dellen formation because of focal corneal dehydration as the result of an altered tear meniscus. PMID- 10826767 TI - Localized neurofibroma of the orbit: a radiographic and histopathologic study. AB - PURPOSE: To demonstrate the clinical, radiographic, intraoperative and histopathologic presentation of a localized neurofibroma of the orbit. METHODS: Clinicopathologic case report. RESULTS: A 25-year-old man was examined for inferior displacement of the right globe. Physical examination and neuro-imaging identified a localized solid tissue mass of the superior orbit. Histopathologic evaluation after complete surgical resection proved the lesion to be a localized neurofibroma. CONCLUSIONS: The patient's history, clinical presentation, and imaging studies were strongly characteristic for this infrequently encountered orbital lesion. Several imaging features, including multilobulation, multiplicity, ring-configured contrast enhancement, and magnetic resonance imaging (MRI) signal intensity heterogeneity, when present in combination, may be reasonably diagnostic of this tumor. PMID- 10826768 TI - Inhibition of proliferation of contaminating fibroblasts by D-valine in cultures of smooth muscle cells from human myometrium. AB - Replacement of L-valine with D-valine in a standard culture medium can selectively inhibit fibroblast proliferation. The aim of the present study was to investigate whether human myometrial cells cultured with D-valine instead of L valine can survive and express their characteristics. Cultured cells (95-98%) maintain expression of the intermediate filament desmin, which is the specific marker for mature muscle cells. By transmission electron microscopy, the cells showed the general morphology of smooth muscle cells in culture. Oxytocin in serum-free culture medium at 37 degrees C (5 min) caused a concentration dependent increase in cellular Na and total Ca, and a decrease in K content as determined by X-ray microanalysis. The percentage of cells cultured with D-valine responding to oxytocin stimulation was larger than that of cells cultured with L valine, suggesting less contamination of smooth muscle cells by fibroblasts in the presence of D-valine. As shown by measurements with fura-2, D-valine-cultured cells retained the characteristic increase in intracellular free Ca2+ ions after oxytocin stimulation. PMID- 10826769 TI - Hydrogen peroxide induces apoptosis in human hepatoma cells and alters cell redox status. AB - Direct exposure of human hepatoma cell line SMMC-7721 to hydrogen peroxide (H2O2) can induce apoptosis. Apoptosis induced by H2O2 was inhibited by cycloheximide, actinomycin D, 3-aminobenzamide, EGTA or Zn2+. H2O2 can increase the level of intracellular Ca2+, downregulate GSH levels, slightly induce lipid peroxidation, and lead to change in the ratio of reduced ion components to oxidized ion components of cells. Analysis of flow cytometry indicates that H2O2 decreases the level of Bcl-2. The data indicate that H2O2-induced apoptosis requires new mRNA and protein syntheses; H2O2 can activate Ca2+/Mg2+-dependent endonuclease leading to internucleosomal DNA fragmentation and activation of poly (ADP-ribose) polymerase interfering with the energy metabolism of the cell. The H2O2 downregulation of GSH may be more important for apoptosis than H2O2 induction of lipid peroxidation, and the H2O2 induced changes in redox status of the cell may be among the original events which lead up to other biochemical changes. PMID- 10826770 TI - Tissues formed during distraction osteogenesis in the rabbit are determined by the distraction rate: localization of the cells that express the mRNAs and the distribution of types I and II collagens. AB - An experimental model of leg lengthening was used to study the morphology of, the collagenous proteins present, and the collagen genes expressed in the regenerating tissue following 20% lengthening at four different distraction rates. At a distraction rate of 0.3 mm/day (8 weeks distraction), the regenerate consists of intramembranous bone and localized areas of fibrocartilage. At rates of 0.7 (4 weeks) and 1.3 mm/day (2 weeks), the bone that grows from the cut ends of the cortical bone is separated by fibrous tissue and cartilage is present. At 2.7 mm/day (1 week), only fibrous tissue and sparse bone are present. Type I collagen is present in the matrices around the cells expressing its mRNA and similarly, type II collagen is located around the chondrocytes. Type I collagen mRNA is expressed predominantly by the fibroblasts in the fibrous tissue, the bone surface cells and to a reduced extent by the osteocytes. Type II collagen mRNA is expressed by chondrocytes. The results suggest that osteoblasts and chondrocytes within the regenerate originate from the same pool of progenitor cells, and the differentiation of these cells and the expression of types I and II collagen genes are altered by different rates of distraction. These observations suggest that the optimal rate of distraction in the model is 0.7 mm/day. PMID- 10826771 TI - S100 immunoreactivity is increased in reactive astrocytes of the visual pathways following a mechanical lesion of the rat occipital cortex. AB - After demonstration of the paracrine action of glial neurotrophic factors, gliosis has also been considered to be related to neuronal trophism and plasticity rather than solely a repair event following brain injury. S100 is a Ca2+ binding protein, present mainly in astrocytes, that exerts paracrine trophic effects on several neuronal populations. This study analyses the presence of S100 protein by means of immunohistochemistry combined with stereology in the reactive glial cells of the rat visual pathways following a lesion of the visual cortex. Adult male Wistar rats were submitted to a unilateral aspiration of the occipital cortex or to a sham operation. One week later the rats were killed and their brain processed for immunochemistry. Single antibody immunohistochemistry was performed for the visualization of glial fibrillary acidic protein (GFAP, a marker for astrocytes), OX-42 (a marker for microglia) and S100 protein. Double immunofluorescence procedures were applied for co-localization of the S100/GFAP and S100/OX-42. An optical dissector, point interceptors and rotators were used to quantify the degree of glial activation and the changes in the S100 immunoreactivity. We observed an intense microglial and astroglial reaction in addition to an increased S100 immunoreactivity in the occipital cerebral cortex, geniculate nucleus and hippocampus ipsilateral to the lesion. In the ipsilateral superior colliculus, an intense astroglial activation was accompanied by an up regulation of S100 immunoreactivity. Double-immunofluoresence revealed an increased S100 immunoreactivity in reactive astrocytes, but not in the reactive microglia. Evidence has therefore been obtained that after mechanical trauma, the astroglial S100 protein participates in the trophism and plasticity of the injured visual pathways. PMID- 10826772 TI - A glycoprotein isolated from the sponge, Pachymatisma johnstonii, has anti leishmanial activity. AB - A high anti-leishmanial activity was observed in an aqueous extract from the marine sponge Pachymatisma johnstonii, Bowerbank 1842 (Demospongiae, Geodiidae). Pachymatismin, a glycoprotein, was purified and shown to be a cytotoxic agent, which acts on promastigote and clinical-like amastigote stages with IC50 about 1 microg protein/ml and induces changes in the cell shape, phospholipase A2 activity and invasion capacity of the parasite. We believe pachymatismin is the first reported substance from a marine organism with anti-leishmanial activity. PMID- 10826773 TI - On the stall force for growing microtubules. AB - The assembly of microtubules generates forces that play a role in cellular motility processes such as the motion of chromosomes during mitosis. Recently, Mogilner and Oster proposed a model for the growth of microtubules that agrees quantitatively with the force-velocity relation measured for individual microtubules. In addition, the authors predicted that the stall force for any polymer consisting of N independently growing protofilaments should increase as the square root of N. We simulated this model and found that the stall force increases linearly with N, and is in fact consistent with the maximum force predicted by thermodynamic arguments. We show that this discrepancy can be explained by a more careful treatment of the "off-term" in the Mogilner-Oster model. PMID- 10826774 TI - A new method for protein domain recognition. AB - A fuzzy cluster method is presented to recognize protein domains. This algorithm can identify domains globally. A protein structure set was used to test the algorithm. Among 219 proteins, 66.7% yielded results that agreed with the reference definitions, 30.6% showed minor differences, and only 2.7% (six proteins) showed major differences with the reference. The new method is more than 20 times fast than previous algorithms. PMID- 10826775 TI - Protein-induced bilayer deformations: the lipid tilt degree of freedom. AB - The theory of hydrophobic interaction between a transmembrane protein and a lipid bilayer is reinvestigated. The protein is modeled as a cylindrically symmetric rigid inclusion, residing in a symmetric, tension-free lipid bilayer. The hydrophobic coupling between the inclusion and the lipids may induce an elastic bilayer deformation, which is commonly described in terms of stretching (or compressing) the hydrocarbon chains of the lipids. In the present work, we additionally include the possibility of the average lipid director to tilt with respect to the normal direction of the hydrocarbon-water interface. The corresponding membrane deformation energy is expressed using both a phenomenological description of elastic lipid layer perturbations and employing a specific molecular lipid model. The molecular lipid model accounts for head group repulsions, interfacial tension, and the chain conformational free energy. Assuming incompressibility of the hydrocarbon chains, we estimate and compare typical membrane deformation energies induced by single gramicidin A channels, with and without the lipid tilt degree of freedom taken into account. The membrane deformation energies are conveniently expressed using a spring constant. We argue that the consideration of the lipid tilt degree of freedom leads to a severalfold reduction of the spring constant and should thus not be excluded from the description of protein-induced membrane deformations. Possible limits of membrane elasticity-based theories for lipid-protein interactions are discussed. Finally, we calculate inclusion-induced deformations of electrostatically charged bilayers, illuminating the coupling between electrostatic and elastic energies in charged membranes. PMID- 10826776 TI - The nicotinic acetylcholine receptor: from molecular model to single-channel conductance. AB - The nicotinic acetylcholine receptor (nAChR) is the archetypal ligand-gated ion channel. A model of the alpha7 homopentameric nAChR is described in which the pore-lining M2 helix bundle is treated atomistically and the remainder of the molecule is treated as a "low resolution" cylinder. The surface charge on the cylinder is derived from the distribution of charged amino acids in the amino acid sequence (excluding the M2 segments). This model is explored in terms of its predicted single-channel properties. Based on electrostatic potential profiles derived from the model, the one-dimensional Poisson-Nernst-Planck equation is used to calculate single-channel current/voltage curves. The predicted single channel conductance is three times higher (ca. 150 pS) than that measured experimentally, and the predicted ion selectivity agrees with the observed cation selectivity of nAChR. Molecular dynamics (MD) simulations are used to estimate the self-diffusion coefficients (D) of water molecules within the channel. In the narrowest region of the pore, D is reduced ca. threefold relative to that of bulk water. Assuming that the diffusion of ions scales with that of water, this yields a revised prediction of the single-channel conductance (ca. 50 pS) in good agreement with the experimental value. We conclude that combining atomistic (MD) and continuum electrostatics calculations is a promising approach to bridging the gap between structure and physiology of ion channels. PMID- 10826777 TI - Tryptophan fluorescence studies of melanotropins in the amphiphile-water interface of reversed micelles. AB - We report studies on the interaction of alpha-melanocyte stimulating hormone (alpha-MSH) and a synthetic analogue (MSH-I) with reverse micelles prepared from the amphiphilic sodium bis(2-ethylhexyl)sulfosuccinate in isooctane. The tripeptide lysyl-tryptophyl-lysine and the isolated amino acid tryptophan were also investigated as simpler compounds interacting with the micelles. Tryptophan fluorescence parameters (spectral position of emission band, anisotropy, and lifetime decay) demonstrated that in the presence of reverse micelles the environment around the fluorophore is less polar and more rigid than bulk water. Those parameters are sensitive to the changes induced in the micelles by the presence of water. In large micelles having a water/ amphiphile molar ratio above 10, the modifications detected by fluorescence are small and the location of the fluorophore is not affected by a further increase in the concentration of the bulk water. The results, with additional support from quenching experiments, indicated that the different compounds occupy different positions in the large reverse micelles, but in any case they are in the interface region, without dispersing into the bulk water. From decay associated spectra, conformations were identified showing different degrees of tryptophan exposition to polar and nonpolar local environments. The conformation related to the long lifetime has its tryptophan more exposed to water while that associated to the intermediate lifetime has that residue stabilized in nonpolar media. The native hormone alpha MSH and the analogue MSH-I present similar conformations in dry micelles. However, in buffer and in the large hydrated micelles, differences in conformations are evident, and could be related to the different physiological activity of the peptides. PMID- 10826778 TI - Circular and linear DNA molecules in the Entamoeba histolytica complex molecular karyotype. AB - Entamoeba histolytica genome was analysed by pulsed field gel electrophoresis under conditions to separate linear chromosomes in the 170-1400 kb range. We identified linear DNA molecules of 227, 366, 631, 850, 1112 and 1361 kb (mean sizes obtained by three different methods) and we estimated their reorientation times and migration velocities at various experimental conditions. DNA shift mobility assays, using ethidium bromide, suggested that bands migrating at 227 and 631 kb contain linear and circular DNA, whereas a band at 436 kb has only circular DNA. We obtained a regression equation relating sizes of supercoiled DNA molecules with their migration velocities during a pulse at constant electric field and temperature. We also developed a computer program (EHPATTERNS) that predicts the migration per pulse and the resolution order of circular and linear E. histolytica DNA at different pulse times and constant driving and frictional forces. The simulation showed that linear DNA molecules frequently co-migrate with circular molecules, but circular molecules change when the pulse time varies. This molecular mixture generates broad bands and difficulties in the interpretation of the molecular karyotype of E. histolytica. PMID- 10826779 TI - Localisation and dynamics of sodium counterions around DNA in solution from molecular dynamics simulation. AB - The localisation and dynamics of sodium counterions around the DNA duplex d(AGCGTACTAGTACGCT)2 corresponding to the trp operator fragment used in the crystal structure of the half site complex has been studied by a 1.4 ns molecular dynamics simulation in explicit solvent. A continuous and well-defined counterion density is shown to be present around the minor groove, while density patches are found in the major groove in regions where DNA bending is observed. A residence time analysis reveals the dynamic nature of these distributions. The resulting picture agrees with previous theoretical and experimental studies of A-tract DNA sequences, and is consistent with the polyelectrolyte condensation model. PMID- 10826780 TI - Steady-state gel electrophoresis of long polymer molecules: a theoretical study. AB - The velocity of long polymer molecules in a gel and the liquid flow profile in the vicinity of a molecule's surface were studied theoretically by combining the Navier-Stokes equation with the Poisson-Boltzmann equation. The electrophoretic mobility has been calculated in dependence of the ionic strength of the electrolyte solution, its viscosity, the gels' volume friction coefficient, the surface charge and the radius of the polymer molecule. The results are presented in a nondimensional form and depend on two dimensionless parameters only. The first parameter is the radius of the polymer molecule in units of the Debye length. The second is a parameter comprising the electrolyte's viscosity and the gel density. Thus, by similarity theory, the results apply to any given experimental arrangement. PMID- 10826781 TI - Validation of publication of new names and new combinations previously effectively published outside the IJSEM. PMID- 10826782 TI - Pseudomonas brassicacearum sp. nov. and Pseudomonas thivervalensis sp. nov., two root-associated bacteria isolated from Brassica napus and Arabidopsis thaliana. AB - Bacteria isolates phenotypically related to Pseudomonas corrugata have frequently been isolated from the rhizosphere of Arabidopsis thaliana and Brassica napus grown on different soils. 16S rDNA (rrs) gene sequencing, DNA-DNA hybridization, biochemical characterization and siderophore typing showed that these isolates belong to two different species that are distinct from other species of the genus Pseudomonas, including P. corrugata. A description of properties of these two new species is given based on the study of 16 isolates. Proposed names are Pseudomonas brassicacearum (10 strains studied) and Pseudomonas thivervalensis (6 strains studied). The type strain of Pseudomonas brassicacearum is CFBP 11706T and that of Pseudomonas thivervalensis is CFBP 11261T. PMID- 10826783 TI - Description of Paralactobacillus selangorensis gen. nov., sp. nov., a new lactic acid bacterium isolated from chili bo, a Malaysian food ingredient. AB - Paralactobacillus selangorensis gen. nov., sp. nov. is described. This organism, isolated from a Malaysian food ingredient called chili bo, is an obligatory homofermentative, rod-shaped lactic acid bacterium. The G+C content is 46.1 46.2+/-0.3 mol%. Earlier 16S rRNA studies showed that this organism constitutes a new taxon distantly related to the Lactobacillus casei-Pediococcus group. A phenotypic description that distinguishes Paralactobacillus selangorensis from other genera of lactic acid bacteria is presented. The type strain of Paralactobacillus selangorensis is LMG 17710T. PMID- 10826784 TI - Desulfotomaculum alkaliphilum sp. nov., a new alkaliphilic, moderately thermophilic, sulfate-reducing bacterium. AB - A new moderately thermophilic, alkaliphilic, sulfate-reducing, chemolithoheterotrophic bacterium, strain S1T, was isolated from a mixed cow/pig manure with neutral pH. The bacterium is an obligately anaerobic, non-motile, Gram-positive, spore-forming curved rod growing within a pH range of 8.0-9.15 (optimal growth at pH 8.6-8.7) and temperature range of 30-58 degrees C (optimal growth at 50-55 degrees C). The optimum NaCl concentration for growth is 0.1%. Strain S1T is an obligately carbonate-dependent alkaliphile. The G+C content of the DNA is 40.9 mol%. A limited number of compounds are utilized as electron donors, including H2+acetate, formate, ethanol, lactate and pyruvate. Sulfate, sulfite and thiosulfate, but not sulfur or nitrate, can be used as electron acceptors. Strain S1T is able to utilize acetate or yeast extract as sources of carbon. Analysis of the 16S rDNA sequence allowed strain S1T (= DSM 12257T) to be classified as a representative of a new species of the genus Desulfotomaculum, Desulfotomaculum alkaliphilum sp. nov. PMID- 10826785 TI - Description of Sphingomonas xenophaga sp. nov. for strains BN6T and N,N which degrade xenobiotic aromatic compounds. AB - The taxonomic position of two bacterial strains, BN6T and N,N, with the ability to degrade xenobiotic aromatic compounds (naphthalenesulfonates or N,N dimethylaniline) was investigated. The 165 rRNA gene sequence, the G+C content of the DNA (62-63 mol%) and the detection of ubiquinone Q-10, 2-hydroxymyristic acid and the sphingoglycolipid present clearly placed the two strains into the genus Sphingomonas. Both strains are representatives of one species according to the level of DNA relatedness (70.7%). The strains could be separated from all validly described taxa of the genus Sphingomonas, according to the 16S rRNA gene sequence (the highest sequence similarity observed was 96 % to Sphingomonas yanoikuyae), the pattern of the polar lipids and physiological characteristics. Therefore, the new species Sphingomonas xenophaga is proposed to accommodate strains BN6T (= DSM 6383T) and N,N (= DSM 8566). PMID- 10826786 TI - Phylogenetic analysis of 18 thermophilic Methanobacterium isolates supports the proposals to create a new genus, Methanothermobacter gen. nov., and to reclassify several isolates in three species, Methanothermobacter thermautotrophicus comb. nov., Methanothermobacter wolfeii comb. nov., and Methanothermobacter marburgensis sp. nov. AB - Using a combination of 16S rRNA analysis and antigenic fingerprinting consisting of new and published data, the phylogenetic position of 18 thermophilic isolates currently classified as Methanobacterium species was reinvestigated. The results were verified by independent methods, including, where applicable, plasmid and phage typing. Comparative analysis of 16S rRNA data for 30 strains belonging to the order Methanobacteriales strongly suggested that mesophilic and thermophilic Methanobacterium isolates are distantly related and should be assigned to separate genera. For the thermophilic strains the genus Methanothermobacter was initially proposed by Boone, Whitman and Rouviere. Furthermore, the results support a reclassification of 15 isolates in three species within the proposed genus: (i) Methanothermobacter thermautotrophicus comb. nov., containing eight isolates, six of which are able to utilize formate (type strain deltaHT); (ii) Methanothermobacter wolfeii comb. nov., containing four formate-utilizing isolates (type strain DSM 2970T); (iii) Methanothermobacter marburgensis sp. nov., containing three obligately autotrophic isolates (type strain MarburgT). Of the nine isolates formerly referred to as Methanobacterium thermoformicicum, six were reclassified as Methanothermobacter thermautotrophicus and three as Methanothermobacter wolfeii. PMID- 10826787 TI - A new species of oral Streptococcus isolated from Sprague-Dawley rats, Streptococcus orisratti sp. nov. AB - Taxonomic studies were performed on an unusual oral Streptococcus strain isolated from Sprague-Dawley rats. The isolates were alpha-haemolytic, bile-tolerant, aesculin-hydrolytic and unable to grow in 6.5% NaCl. They fermented lactose, sucrose and trehalose. They were distinguished from other recognized species of oral and viridans streptococci by several biochemical characteristics and by Lancefield's group antigen, as well as by unique DNA-DNA hybridization characteristics. 16S rDNA sequence studies confirmed the genealogical distinctiveness of the species. The results of the study demonstrated that the isolates represented a new species of the oral and viridans streptococci. The name Streptococcus orisratti sp. nov. is proposed for the new species. The type strain is A63T (= ATCC 700640T). PMID- 10826788 TI - 'Candidatus Odyssella thessalonicensis' gen. nov., sp. nov., an obligate intracellular parasite of Acanthamoeba species. AB - An intracellular bacterium, strain L13, was observed infecting an environmental isolate of an Acanthamoeba species. The bacterium could not be recovered on axenic medium but was recovered and cultivated in vitro using cultures of Acanthamoeba polyphaga. The 16S rRNA gene sequence of L13 was found to be new, sharing less than 84% similarity with other sequences in the GenBank/EMBL database. L13 was found to be a member of the alpha-Proteobacteria, sharing an evolutionary line of descent with a group of uniquely obligate intracellular organisms comprised of Caedibacter and Holospora species and the NHP bacterium. Viable bacteria appeared to be highly motile within amoebae. Ultrastructural analysis of the bacterium demonstrated that it is rod-shaped and possesses a typical Gram-negative cell wall, but has no other outstanding features except small vesicle-like structures often associated with the outer surface of each bacterium. The host range of L13 was found to be limited to the genus Acanthamoeba. In A. polyphaga, L13 infection was slow to manifest when cultures were incubated below 30 degrees C, but at higher temperatures bacteria multiplied prolifically and induced host cell lysis. The protein profile of the bacterium purified from the amoebae was assessed by SDS-PAGE and its G+C content was estimated to be 41 mol%. Although these results support the proposal of L13 as a new species, its obligate intracellular nature prevented isolation of a definitive type strain. L13 is therefore proposed as 'Candidatus Odyssella thessalonicensis' gen. nov., sp. nov. PMID- 10826789 TI - Nocardiopsis tropica sp. nov., Nocardiopsis trehalosi sp. nov., nom. rev. and Nocardiopsis dassonvillei subsp. albirubida subsp. nov., comb. nov. AB - Sixteen strains isolated from different soils and marine sediments and 'Nocardiopsis trehalosi' VKM Ac-942T were subjected to taxonomic analyses. The morphological and chemotaxonomic characteristics of the strains matched those described for members of the Nocardiopsis. Based on 16S rDNA sequence analysis, DNA-DNA hybridization values and phenotypic characteristics, including the composition of menaquinones and cell wall teichoic acids, two new species are proposed, Nocardiopsis tropica sp. nov. (type strain VKM Ac-1457T) and Nocardiopsis trehalosi sp. nov., nom. rev. (type strain VKM Ac-942T = JCM 3357T = NRRL 12026T). Nocardiopsis dassonvillei VKM Ac-1882T (formerly Nocardiopsis albirubida) differs from other strains of this species by the presence of MK-10 as the major menaquinone, cell wall teichoic acid structure, and numerical analysis based on 115 growth and physiological characteristics. The strain is reclassified as the subspecies Nocardiopsis dassonvillei subsp. albirubida, with VKM Ac-1882T (= DSM 43465T = ATCC 23612T = ISP 5465T) designated the type. PMID- 10826790 TI - Pseudomonas plecoglossicida sp. nov., the causative agent of bacterial haemorrhagic ascites of ayu, Plecoglossus altivelis. AB - A new Pseudomonas species, for which the name Pseudomonas plecoglossicida is proposed, was isolated from cultured ayu (Plecoglossus altivelis) with bacterial haemorrhagic ascites. The causative agent was similar to Pseudomonas putida biovar A in its phenotypic characteristics and on the basis of 16S rRNA gene sequence analysis, but it reduced nitrate to nitrite. Furthermore, it was distinguished phenotypically from Pseudomonas putida biovar A by utilization of D malate, L-(+)-tartrate, m-tartrate and nicotinate. The levels of DNA-DNA hybridization between the isolate strain FPC 951T and other reference strains of Pseudomonas species, including Pseudomonas putida, were less than 50%. The G+C content of the DNA of FPC 951T was 62.8 mol%. Strain FPC 951T (= ATCC 700383T) is designated the type strain of the new species. PMID- 10826791 TI - DNA relatedness among strains of Streptomyces pathogenic to potato in France: description of three new species, S. europaeiscabiei sp. nov. and S. stelliscabiei sp. nov. associated with common scab, and S. reticuliscabiei sp. nov. associated with netted scab. AB - The genomic relatedness was evaluated by DNA-DNA hybridization for 23 strains (21 were pathogenic and two were saprophytic strains) isolated from lesions of common and netted scab in France and 19 strains from other countries, including type strains of Streptomyces species. Three genomospecies were defined within the conventional species of Streptomyces scabies, and these genomospecies were different from other pathogenic described species (Streptomyces acidiscabies, Streptomyces caviscabies) based on previously published phenotypic data. Two of these genomospecies (1 and 3) correspond to new species, for which the names Streptomyces europaeiscabiei sp. nov. (with type strain CFBP 4497T) and Streptomyces stelliscabiei sp. nov. (with type strain CFBP 4521T) are proposed. Genomospecies 2 corresponds to S. scabies (with type strain CFBP 4517T = ATCC 49173T), and includes only one French strain. The pathogenic strains associated with netted scab lesions constituted a new species that was named Streptomyces reticuliscabiei sp. nov. (with type strain CFBP 4531T). The G+C content of DNA from the three strains CFBP 4497T (S. europaeiscabiei), CFBP 4521T (S. stelliscabiei), CFBP 4531T (S. reticuliscabiei) was 71.3, 71.0 and 69.8 mol%, respectively. Phylogenetic analysis based on 16S rRNA gene sequences showed that the type strain CFBP 4497T was very similar to the type strain of S. scabies, whereas, the type strain of S. stelliscabiei, CFBP 4521T, was very similar to the type strain of Streptomyces bottropensis. On the basis of 16S rRNA gene sequences, the type strain of S. reticuliscabiei, CFBP 4531T, differed extensively from the other strains of Streptomyces tested. PMID- 10826792 TI - Reclassification of Clostridium quercicolum as Dendrosporobacter quercicolus gen. nov., comb. nov. AB - Morphological features, genomic DNA base composition and 16S rDNA sequence similarities, as well as a distinct phospholipid pattern, whole-cell fatty acid distribution and the occurrence of the lipoquinone 'lipid F', indicate that Clostridium quercicolum belongs to the Sporomusa-Pectinatus-Selenomonas phyletic group and possesses only a remote relationship to members of the genus Clostridium sensu stricto. On the basis of these results, the new genus and combination Dendrosporobacter quercicolus gen. nov., comb. nov. are proposed. PMID- 10826793 TI - Clostridium gasigenes sp. nov., a psychrophile causing spoilage of vacuum-packed meat. AB - Two psychrophilic Clostridium strains, DB1AT and R26, were isolated from incidences of 'blown-pack' spoilage of vacuum-packed chilled lamb. Vacuum packs of meat inoculated with these strains developed gas bubbles and pack distension within 14 d storage at 2 degrees C. The two main gases responsible for pack distension were carbon dioxide and hydrogen. 1-Butanol, butyric and acetic acid and butyl esters were the major volatile compounds produced by the strains in the artificially inoculated packs. The unknown strains were Gram-positive motile rods producing elliptical subterminal spores during the late-stationary growth phase. At pH 7.0, they grew from -1.5 to 26 degrees C, and their optimum growth temperature was 20-22 degrees C. At 20 degrees C, the pH range for growth was 5.4 8.9 and the optimum pH for growth was 6.2-8.6. In peptone/yeast extract broth, the organisms grew little or not at all in the absence of fermentable carbohydrates. Both strains hydrolysed gelatin, aesculin and starch. The fermentation products formed in peptone yeast extract glucose starch broth were ethanol, acetate, butyrate, lactate, butanol, carbon dioxide and hydrogen. The G+C contents of the DNA of strains DB1AT and R26 were 29.4 and 28.3 mol%, respectively. Phylogenetic analyses indicated that the strains belong to cluster I of the genus Clostridium (sensu Collins et al. 1994). The new strains differed from the phylogenetically related clostridia in cellular fatty acid composition, soluble protein profiles and phenotypic properties. On the basis of rDNA analysis and phenotypic and phylogenetic characterization, the strains were assigned to a new species for which the name Clostridium gasigenes is proposed. Strain DB1AT (= DSM 12272T) is designated as the type strain. PMID- 10826794 TI - Bifidobacterium thermacidophilum sp. nov., isolated from an anaerobic digester. AB - A new phenotypic group of Bifidobacterium strains, isolated from an anaerobic digester for the treatment of waste water from a bean-curd farm, was described previously. In this study, the DNA-DNA relatedness between strain 36 (type strain, AS 1.2282T) of this new group and the type strains of other described Bifidobacterium species was analysed. The low level of DNA homology (0-58.9%) as well as comparison of the 16S rDNA sequences confirmed the distinct phylogenetic position of strain 36. In addition, the new species differed from other Bifidobacterium species in its phenotypic characteristics, such as its growth at moderately thermophilic conditions (49.5 degrees C) and at relatively low pH (4.0), as well as its sugar-fermentation pattern. On the basis of phenotypic, genetic and phylogenetic studies, a new Bifidobacterium species, Bifidobacterium thermacidophilum sp. nov., was designated. PMID- 10826795 TI - Intrageneric relationships among Micromonospora species deduced from gyrB-based phylogeny and DNA relatedness. AB - The phylogenetic structure of genus Micromonospora within actinomycetes was examined by analysing the gyrB sequences of 15 validly described species and four subspecies. All but one of the Micromonospora strains formed a tight cluster, as had previously been demonstrated by a 16S rDNA-based phylogenetic analysis. However, the intrageneric relationships deduced from the gyrB-based phylogeny were different from those based on their 16S rDNA sequences. To examine which phylogeny would be more relevant for classifying genus Micromonospora, DNA-DNA hybridization experiments were performed. The gyrB-based classification agrees with the results of the DNA-DNA hybridization studies, indicating that this classification method is useful for analysing the phylogenetic relationships of high G+C Gram-positive bacteria at the level of the genomic species. Genus Micromonospora was reclassified into the following 14 species: Micromonospora echinospora, Micromonospora pallida, Micromonospora nigra, Micromonospora purpureochromogenes, Micromonospora aurantiaca, Micromonospora carbonacea, Micromonospora chalcea, Micromonospora chersina, Micromonospora coerulea, Micromonospora gallica, Micromonospora halophytica, Micromonospora inositola, Micromonospora olivasterospora and Micromonospora rosaria. PMID- 10826796 TI - Phocoenobacter uteri gen. nov., sp. nov., a new member of the family Pasteurellaceae Pohl (1979) 1981 isolated from a harbour porpoise (Phocoena phocoena). AB - Phenotypic and phylogenetic studies were performed on a Gram-negative, rod-shaped bacterium isolated from the uterus of a porpoise. Biochemical and physiological studies indicated that the bacterium was related to the family Pasteurellaceae. Comparative 16S rRNA gene sequencing studies confirmed these findings and demonstrated that the bacterium represents a hitherto unknown subline within this family of organisms. Based on the results of the phylogenetic analysis and phenotypic criteria, it is proposed that the bacterium be assigned to a new genus, Phocoenobacter uteri gen. nov., sp. nov. The type strain of Phocoenobacter uteri sp. nov. is NCTC 12872T. PMID- 10826797 TI - Reclassification of Alteromonas distincta Romanenko et al. 1995 as Pseudoalteromonas distincta comb. nov. AB - The 16S rRNA gene of Alteromonas distincta KMM 638T was amplified, cloned and sequenced. The nucleotide sequence was aligned with sequences of representative strains of Alteromonas, Moritella, Pseudoalteromonas and Shewanella. Results of phylogenetic analysis, using neighbour-joining and Fitch-Margoliash methods, clearly indicated that this species should be assigned to the genus Pseudoalteromonas. On the basis of polyphasic data obtained from previous work and this study, it is proposed that the species Alteromonas distincta be reclassified as Pseudoalteromonas distincta comb. nov. with type strain KMM 638T (= ATCC 700518T). PMID- 10826798 TI - Bacteroides acidifaciens sp. nov., isolated from the caecum of mice. AB - During studies of the bacterial flora in the intestines of mice, we isolated characteristic strains which lowered the pH of peptone-yeast broth containing Fildes' digest. Based on 16S rRNA sequence comparison, these isolates were considered to belong to the Bacteroides cluster in the bacteroides subgroup of the Cytophaga-Flavobacterium-Bacteroides phylum, and were divided into two groups. Their phenotypic characteristics, i.e. growth in 20% bile, aesculin hydrolysis, and glucose-6-phosphate dehydrogenase (G6PDH) and 6-phosphogluconate dehydrogenase (6PGDH) activity, were the same as those of the 'Bacteroides fragilis group'. The low level of DNA-DNA hybridization with type strains in the Bacteroides cluster confirmed the novel species status of these isolates. It is proposed that these isolates be named Bacteroides acidifaciens, the type strain of which is A40T (= JCM 10556T). PMID- 10826799 TI - Characterization of the rnpB gene and RNase P RNA in the order Chlamydiales. AB - The sequence of the RNase P RNA gene (rnpB) was determined for 60 strains representing all nine species in the family Chlamydiaceae and for the related Chlamydiales species, Parachlamydia acanthamoebae and Simkania negevensis. These sequences were used to infer evolutionary relationships among the Chlamydiaceae. The analysis separated Chlamydophila and Chlamydia into two lineages, with Chlamydophila forming three distinct clusters: the Chlamydophila pneumoniae strains; the Chlamydophila pecorum strains; and a third cluster comprising the species Chlamydophila psittaci, Chlamydophila abortus, Chlamydophila caviae and Chlamydophila felis. The Chlamydia line of descent contained two clusters, with the Chlamydia suis strains distinctly separated from strains of Chlamydia trachomatis and Chlamydia muridarum. This analysis indicated that the rnpB sequence and structure are distinctive markers for species in the Chlamydiaceae. It was also demonstrated that the RNase P RNA derived from Chlamydia trachomatis is able to cleave a tRNA precursor in the absence of protein. These findings are discussed in relation to the structure of Chlamydia RNase P RNA. PMID- 10826800 TI - Polyphasic characterization of rhizobia that nodulate Phaseolus vulgaris in West Africa (Senegal and Gambia). AB - Fifty-eight new isolates were obtained from root nodules of common bean (Phaseolus vulgaris) cultivated in soils originating from different agroecological areas in Senegal and Gambia (West Africa). A polyphasic approach including both phenotypic and genotypic techniques was used to study the diversity of the 58 Rhizobium isolates and to determine their taxonomic relationships with reference strains. All the techniques performed, analysis of multilocus enzyme electrophoretic patterns, SDS-PAGE profiles of total cell proteins, PCR-RFLP analysis of the genes encoding 16S rRNA and of the 16S-23S RNA intergenic spacer region (ITS-PCR-RFLP), auxanographic tests using API galleries and nodulation tests lead to the consensus conclusion that the new rhizobial isolates formed two main distinct groups, I and II, belonging to Rhizobium tropici type B and Rhizobium etli, respectively. By MLEE R. etli and group II strains showed several related electrophoretic types, evidencing some extent of internal heterogeneity among them. This heterogeneity was confirmed by other techniques (ITS-PCR-RFLP, SDS-PAGE and host-plant-specificity) with the same nine distinct strains of group II showing some differences from the core of group II (54 strains). PMID- 10826801 TI - Methanosarcina semesiae sp. nov., a dimethylsulfide-utilizing methanogen from mangrove sediment. AB - Methanosarcina semesiae MD1T (T = type strain), a novel obligately methylotrophic methanogenic archaeon is described. Strain MD1T was isolated from an enrichment on dimethylsulfide inoculated with mangrove sediment. The cells were irregularly coccoid, non-motile, 1.4+/-0.2 microm in diameter and stained Gram-positive. The catabolic substrates used included dimethylsulfide, methanethiol, methanol and methylated amines, but not acetate, formate, H2/CO2 or a combination of these substrates. When cells grown on dimethylsulfide were transferred to trimethylamine or methanol and vice versa, a lag phase was observed. The same lag phase occurred when cells grown on trimethylamine were transferred to methanol and vice versa, indicating that for each substrate different enzymes were induced. Fastest growth occurred within a temperature range of 30-35 degrees C and a pH of 6.5-7.5. Both Na+ and Mg2+ were required for growth, with maximum growth rates at 200-600 mM Na+ and 20-100 mM Mg2+. The cells exhibited specific growth rates (h-1) of 0.07+/-0.02, 0.15+/-0.04 and 0.18-/+0.05 on dimethylsulfide, methanol and trimethylamine, respectively. Analysis of the 16S rRNA gene sequence showed that strain MD1T was phylogenetically closely related to members of the genus Methanosarcina, but clearly differed from all described species of this genus (94-97% sequence similarity). PMID- 10826802 TI - Luteococcus peritonei sp. nov., isolated from the human peritoneum. AB - An unusual catalase-positive pleomorphic Gram-positive rod isolated from a human clinical specimen was subjected to a polyphasic taxonomic analysis. Comparative 16S rRNA gene sequence analysis revealed the unknown bacterium was a member of the high G+C branch of the Gram-positive bacteria (Actinobacteria), and was phylogenetically a member of the family Propionibacteriaceae, with Luteococcus japonicus as its nearest relative. Based on both phenotypic and phylogenetic evidence, it is proposed that the unknown bacterium be classified in the genus Luteococcus, as Luteococcus peritonei sp. nov. The type strain of Luteococcus peritonei is CCUG 38120T. PMID- 10826803 TI - Halorhabdus utahensis gen. nov., sp. nov., an aerobic, extremely halophilic member of the Archaea from Great Salt Lake, Utah. AB - Strain AX-2T (T = type strain) was isolated from sediment of Great Salt Lake, Utah, USA. Optimal salinity for growth was 27% (w/v) NaCl and only a few carbohydrates supported growth of the strain. Strain AX-2T did not grow on complex substrates such as yeast extract or peptone. 16S rRNA analysis revealed that strain AX-2T was a member of the phyletic group defined by the family Halobacteriaceae, but there was a low degree of similarity to other members of this family. The polar lipid composition comprising phosphatidyl glycerol, the methylated derivative of diphosphatidyl glycerol, triglycosyl diethers and sulfated triglycosyl diethers, but not phosphatidyl glycerosulfate, was not identical to that of any other aerobic, halophilic species. On the basis of the data presented, it is proposed that strain AX-2T should be placed in a new taxon, for which the name Halorhabdus utahensis is appropriate. The type strain is strain AX-2T (= DSM 12940T). PMID- 10826804 TI - Hongia gen. nov., a new genus of the order Actinomycetales. AB - An aerobic, nocardioform actinomycete, named LM 161T, was isolated from a soil sample obtained from a gold mine in Kongiu, Republic of Korea. This organism formed well-differentiated aerial and substrate mycelia and produced branched hyphae that fragmented into short or elongated rods. The cell wall contains major amounts of LL-diaminopimelic acid, alanine, glycine, glutamic acid, mannose, glucose, galactose, ribose and acetyl muramic acid. The major phospholipids of this isolate are phosphatidylcholine, diphosphatidylglycerol, phosphatidylglycerol and phosphatidylinositol, and the major isoprenologue is a tetrahydrogenated menaquinone with nine isoprene units. The whole-cell hydrolysate of strain LM 161T contains 12-methyltetradecanoic and 14 methylpentadecanoic acids as the predominant fatty acids, but does not contain mycolic acids. The G+C content of the DNA is 71.3 mol%. The phylogenetic position of the test strain was investigated using an almost complete 16S rDNA sequence. The isolate formed the deepest branch in the clade encompassing the members of the suborder Propionibacterineae Rainey et al. 1997. On the basis of chemical, phenotypic and genealogical data, it is proposed that this isolate be classified within a new genus as Hongia koreensis gen. nov., sp. nov. in the order Actinomycetales. The type strain is LM 161T (= IMSNU 50530T). PMID- 10826805 TI - Micropruina glycogenica gen. nov., sp. nov., a new Gram-positive glycogen accumulating bacterium isolated from activated sludge. AB - A new Gram-positive non-spore-forming bacterium, strain Lg2T, was isolated from an activated sludge reactor showing enhanced biological phosphorus removal activity. The new isolate was a slowly growing organism and was capable of accumulating large amounts of intracellular glycogen from substrate taken up. Both oxidase and catalase were produced. The new isolate contained meso diaminopimelic acid (DAP) in the cell wall. Complex fatty acid patterns with iso C14:0, anteiso-C15:0, C16:0, iso-C16:0 and four other minor saturated or unsaturated straight-chain fatty acids were detected. The isolate contained a high genomic G+C content (70.5 mol%). Phylogenetic analysis based on the 16S rRNA gene sequence placed the isolate in the high G+C Gram-positive group with Microlunatus phosphovorus and Friedmanniella antarctica as the closest relatives (sequence similarities are 93 and 92 %, respectively). These three organisms shared common features in morphology, but strain Lg2T could be differentiated from the other species by its peptidoglycan type (meso-DAP), fatty acid composition, carbon source utilization profile and G+C content. On the basis of these findings, it is proposed that a new genus and species, Micropruina glycogenica, should be created for the new isolate; the type strain is strain Lg2T (= JCM 10248T). PMID- 10826806 TI - Thermus igniterrae sp. nov. and Thermus antranikianii sp. nov., two new species from Iceland. AB - Several yellow-pigmented isolates, with optimum growth temperatures of about 65 70 degrees C, were recovered from hot springs in Iceland. Phylogenetic analysis of the 16S rDNA and DNA-DNA reassociation values showed that these organisms represented two new species of the genus Thermus. Strains RF-4T and HN1-8 had maximum temperatures for growth below 80 degrees C, while strains HN3-7T and HN2 7, unlike all other strains of the species of the genus Thermus except those belonging to Thermus thermophilus, grew at 80 degrees C. The new isolates from Iceland could not be distinguished easily from each other or from other strains of the species of the genus Thermus by biochemical characteristics; however, strains RF-4T and HN1-8 assimilated ribitol, a characteristic which was not detected in any of the other strains examined. Moreover, the species represented by strains RF-4T and HN1-8 and the species represented by strains HN3-7T and HN2 7 could be distinguished clearly from the other species of Thermus by their fatty acid composition. Strains RF-4T and HN1-8 have the highest combined levels of iso 15:0 and iso-17:0 and the lowest levels of iso-16:0 of any of the strains of the species of Thermus, while strains HN3-7T and HN2-7 are characterized by a very low iso-15:0/iso-17:0 ratio. On the basis of the phylogenetic analysis, DNA-DNA reassociation values, physiological and biochemical characteristics and fatty acid composition, the name Thermus igniterrae sp. nov. is proposed for the species represented by strains RF-4T and HN1-8 and the name Thermus antranikianii sp. nov. is proposed for the species represented by strains HN3-7T and HN2-7. PMID- 10826807 TI - A proposal for the reclassification of Bdellovibrio stolpii and Bdellovibrio starrii into a new genus, Bacteriovorax gen. nov. as Bacteriovorax stolpii comb. nov. and Bacteriovorax starrii comb. nov., respectively. AB - Bdellovibrios are unique bacteria with the ability to prey upon a wide variety of susceptible Gram-negative bacteria. Micro-organisms exhibiting this trait have been included in the genus Bdellovibrio despite their isolation from diverse habitats and relatively unstudied taxonomic relatedness. In this study, 16S rDNA sequences were compared from known terrestrial Bdellovibrio species, Bdellovibrio bacteriovorus 100T, Bdellovibrio stolpii Uki2T and Bdellovibrio starrii A3.12T in order to study their phylogenetic relationship. The two sequences from B. stolpii Uki2T and B. starrii A3.12T were 90.0% similar to each other but exhibited only 81.7% and 81.2% similarity, respectively to B. bacteriovorus 100T. Phylogenetic analysis indicated that B. bacteriovorus 100T clustered in a separate clade from B. starrii A3.12T and B. stolpii Uki2T, demonstrating only a distant relationship between B. bacteriovorus 100T and the other two recognized type species. DNA-DNA hybridization experiments also demonstrated <4% hybridization between these three species. On the basis of the results obtained from the phylogenetic analysis and DNA-DNA hybridization studies, it is proposed that B. stolpii Uki2T and B. starrii A3.12T should be transferred to a new genus, Bacteriovorax gen. nov. as Bacteriovorax stolpii comb. nov. and Bacteriovorax starrii comb. nov., respectively. It is also proposed that the type species for the new genus Bacteriovorax should be Bacteriovorax stolpii comb. nov. PMID- 10826808 TI - Bradyrhizobium spp. (TGx) isolates nodulating the new soybean cultivars in Africa are diverse and distinct from bradyrhizobia that nodulate North American soybeans. AB - The newly developed cultivars of soybean in Africa, known as Tropical Glycine cross (TGx), are nodulated by bradyrhizobia indigenous to African soils, here designated Bradyrhizobium spp. (TGx). Isolates of Bradyrhizobium spp. (TGx) obtained from nodules of TGx soybeans that were inoculated with soils from 65 locations in six African countries were characterized and grouped into 11 phylogenetic clusters on the basis of RFLP of the 16S rRNA gene. Five restriction enzymes (RsaI, HinfI, MspI, CfoI and HaeIII) established RFLP groups within these Bradyrhizobium spp. (TGx) isolates, which were used to construct a phylogenetic tree showing their genetic relationship with other Bradyrhizobium species. RFLP analysis indicated that Bradyrhizobium spp. (TGx) is a heterogeneous group with some isolates related to Bradyrhizobium japonicum and Bradyrhizobium elkanii strains and some to Bradyrhizobium spp. (misc.) reference strains isolated from a variety of tropical legumes. The heterogeneity within the large phylogenetic clusters was further examined through analysis of randomly amplified polymorphic DNA (RAPD) using GC-rich PCR primers. The RAPD analysis showed additional heterogeneity in the Bradyrhizobium spp. (TGx) phylogenetic clusters, which was not revealed by separations based on RFLP analysis. The Bradyrhizobium spp. (TGx) isolates were classified into effective and ineffective types based on their symbiotic performance on TGx soybean. The isolates were randomly distributed throughout the phylogenetic clusters regardless of their symbiotic effectiveness on TGx soybean. PMID- 10826809 TI - Acidovorax anthurii sp. nov., a new phytopathogenic bacterium which causes bacterial leaf-spot of anthurium. AB - The bacterial leaf-spot of anthurium emerged during the 1980s, in the French West Indies and Trinidad. This new bacterial disease is presently wide spread and constitutes a serious limiting factor for commercial anthurium production. Twenty nine strains isolated from leaf-spots of naturally infected anthurium were characterized and compared with reference strains belonging to the Comamonadaceae family, the genera Ralstonia and Burkholderia, and representative fluorescent pseudomonads. From artificial inoculations 25 out of 29 strains were pathogenic on anthurium. Biochemical and physiological tests, fatty acid analysis, DNA-DNA hybridization, 16S rRNA gene sequence analysis, DNA-16S RNA hybridization were performed. The 25 pathogenic strains on anthurium were clustered in one phenon closely related to phytopathogenic strains of the genus Acidovorax. Anthurium strains were 79-99% (deltaTm range 0.2-1.6) related to the strain CFBP 3232 and constituted a discrete DNA homology group indicating that they belong to the same species. DNA-rRNA hybridization, 16S rRNA sequence and fatty acid analysis confirmed that this new species belongs to the beta-subclass of Proteobacteria and to rRNA superfamily III, to the family of Comamonadaceae and to the genus Acidovorax. The name Acidovorax anthurii is proposed for this new phytopathogenic bacterium. The type strain has been deposited in the Collection Francaise des Bacteries Phytopathogenes as CFBP 3232T. PMID- 10826810 TI - DNA-DNA reassociation studies of Streptococcus constellatus with unusual 16S rRNA sequences. AB - DNA-DNA reassociation studies were performed on previously described 'CI strains', which form an unusual 16S rRNA population within the 'anginosus' group of Streptococcus. The CI strains displayed reassociation values of >70% with the Streptococcus constellatus NCDO 2226T strain, with Tm values <1 degrees C, indicating phylogenetic species identity. PMID- 10826811 TI - Lactobacillus mucosae sp. nov., a new species with in vitro mucus-binding activity isolated from pig intestine. AB - A new Lactobacillus species from pig small intestine has been identified. In an attempt to isolate Lactobacillus reuteri strains carrying the putative colonization-factor gene (mub, for mucus binding) a mub-derived gene probe was used to screen pig intestinal material. A number of isolates were obtained and primary characterization showed that they were Gram-positive, catalase-negative, non-spore-forming, non-motile rods. Growth occurred at 45 degrees C but not at 15 degrees C and the DNA G+C content was 46 mol%. Cell wall analysis together with DNA-DNA hybridization and analysis of the 16S rRNA sequence revealed that the new isolates represent a previously undescribed Lactobacillus species closely related to L. reuteri, Lactobacillus fermentum and Lactobacillus pontis. The name Lactobacillus mucosae is proposed for this species and the type strain is S32T. PMID- 10826812 TI - Aminobacterium mobile sp. nov., a new anaerobic amino-acid-degrading bacterium. AB - A novel, curved (0.3 x 4.0-5.0 microm), Gram-negative, non-sporulating, mesophilic bacterium, designated strain ILE-3T (T = type strain), was isolated from an anaerobic lagoon in a dairy wastewater treatment plant. Optimal growth occurred at 37 degrees C and pH 7.4 on a medium containing serine as an energy source and yeast extract. The strain was motile by means of one or two lateral flagella. It required yeast extract for growth on serine, glycine, threonine and pyruvate. Poor growth was obtained on cysteine, Casamino acids, biotrypcase, peptone and 2-oxoglutarate. In the presence of Methanobacterium formicicum, strain ILE-3T oxidized alanine, glutamate, leucine, isoleucine, valine and aspartate to a minor extent. The G+C content of the DNA was 44 mol%. Phylogenetic analysis of the 16S rRNA gene of strain ILE-3T indicated that it was related to Aminobacterium colombiense (95% similarity value). On the basis of the phenotypic and phylogenetic characteristics, strain ILE-3T is designated as a new species of the genus Aminobacterium, namely Aminobacterium mobile sp. nov. (= DSM 12262T). PMID- 10826813 TI - Assignment of Alteromonas elyakovii KMM 162T and five strains isolated from spot wounded fronds of Laminaria japonica to Pseudoalteromonas elyakovii comb. nov. and the extended description of the species. AB - A marine bacterium, Alteromonas elyakovii KMM 162T, which was described recently, and five strains isolated from spot-wounded fronds of Laminaria japonica have been subjected to phylogenetic analysis, and geno- and phenotypic characterization. The phenotypic features of Pseudoalteromonas elyakovii strains were closely related to that of Pseudoalteromonas espejiana IAM 12640T, but utilization of three carbon compounds (D-mannose, L-tyrosine and trehalose) distinguished both species. The G+C content of Pseudoalteromonas elyakovii was between 38.5 and 38.9 mol%. Pseudoalteromonas elyakovii KMM 162T and the five Laminaria isolates constitute a single species different from any other Alteromonas and Pseudoalteromonas species as revealed by DNA-DNA hybridization data, especially Pseudoalteromonas distincta KMM 638T (52.4%), Pseudoalteromonas citrea KMM 216 (49.5%), Pseudoalteromonas carrageenovora NCIMB 302T (46.9%) and Pseudoalteromonas espejiana IAM 12640T (29.9%). All the data indicated that Alteromonas elyakovii KMM 162T should be reclassified as Pseudoalteromonas elyakovii and five strains isolated from Laminaria japonica have to be included in the species. Pseudoalteromonas elyakovii comb. nov. (type strain, KMM 162T = ATCC 700519T) is proposed and a set of phenotypic features which differentiate the Pseudoalteromonas species is described. PMID- 10826814 TI - Characterization of N2O-producing Xanthomonas-like isolates from biofilters as Stenotrophomonas nitritireducens sp. nov., Luteimonas mephitis gen. nov., sp. nov. and Pseudoxanthomonas broegbernensis gen. nov., sp. nov. AB - A group of yellow-pigmented isolates from ammonia-supplied biofilters showed an unusual denitrification reaction. All strains reduced nitrite but not nitrate without production of nitrogen (N2). The only product found was nitrous oxide (N2O). The strains were divided into two clusters and one separate strain by their fatty acid profiles, which were similar to the fatty acid profiles of the genera Xanthomonas and Stenotrophomonas. Analyses of the 165 rDNA sequences showed that these clusters and the separate strain form three independent lines within the Xanthomonas branch of the Proteobacteria. The evolutionary distances of the isolates to members of the related genera Xanthomonas, Stenotrophomonas and Xylella calculated by the 16S rDNA sequences led to the proposal of two new genera and three new species, Stenotrophomonas nitritireducens sp. nov., Luteimonas mephitis gen. nov., sp. nov. and Pseudoxanthomonas broegbernensis gen. nov., sp. nov. The type strains are Stenotrophomonas nitritireducens L2T (= DSM 12575T), Luteimonas mephitis B1953/27.1T (= DSM 12574T) and Pseudoxanthomonas broegbernensis B1616/1T (= DSM 12573T). PMID- 10826815 TI - Mycobacterium xenopi and related organisms isolated from stream waters in Finland and description of Mycobacterium botniense sp. nov. AB - Three scotochromogenic Mycobacterium xenopi-like organisms were isolated from stream waters in Finland. These strains grew at 36-50 degrees C but not at 30 degrees C. One of the three strains was fully compatible with the M. xenopi type strain according to GLC-MS, biochemical tests, and 16S rDNA and 16S-23S rDNA internal transcribed spacer (ITS) sequencing. Two of the strains closely resembled M. xenopi in lipid analyses and biochemical tests, but analysis by GLC MS verified the presence of two new marker fatty acids (2,4,6,x-tetramethyl eicosanoic acid and 2,4,6,x,x-pentamethyl-docosanoic acid). The 16S rDNA and ITS region sequences of these two strains differed from those of M. xenopi and other previously described mycobacterial sequences. Therefore, the strains are regarded as new species of slow-growing mycobacteria, for which the name Mycobacterium botniense sp. nov. is proposed. The chemical, physical and microbiological quality of the water reservoirs of M. xenopi and M. botniense are described. As far as is known, this is the first time that M. xenopi has been isolated from natural waters. The strains of M. botniense sp. nov. (E347T and E43) have been deposited in the ATCC as strains 700701T and 700702, respectively. PMID- 10826816 TI - Updated phylogenetic description of the Mycoplasma hominis cluster (Weisburg et al. 1989) based on 16S rDNA sequences. AB - The fastidious nature of the mollicutes (mycoplasmas), their lack of a classic bacterial cell wall, and their very small genome, make phylogenetic placements of new species in this enlarging group of prokaryotes an important and valuable aid in their classification. In this report we have determined the phylogeny of the Mycoplasma hominis cluster of the hominis group. The 16S rDNA sequences from several previously described Mycoplasma species were determined and ten species were found to belong to the M. hominis cluster. With almost complete sequences available, the phylogenetic analysis revealed that the M. hominis cluster currently comprises 19 species, forming a distinct clade as judged from branch lengths, bootstrap percentage values, nucleotide signature analysis, and structural elements in the 16S rRNA molecule. The 16S rRNA gene sequences of species in the M. hominis cluster were found to be > or = 94% similar and the range within which similarities can be used in the classification of new species is discussed. Members of the M. hominis cluster all share a major biochemical property of M. hominis, in that they hydrolyse arginine and are incapable of fermenting glucose. This consistency in phenotypic pattern has not been found in any of the other phylogenetic clusters of the hominis group. Two species, the non cultivable agent of Grey Lung disease in rodents (tentatively named 'Candidatus Mycoplasma ravipulmonis') and the avian species Mycoplasma gypis strain B1/T1T, were regarded as close relatives to the M. hominis cluster, but are clearly separated from the species of this cluster. Both species formed early branches of the M. hominis cluster and should be regarded as individual lines containing one species. PMID- 10826817 TI - Staleya guttiformis gen. nov., sp. nov. and Sulfitobacter brevis sp. nov., alpha 3-Proteobacteria from hypersaline, heliothermal and meromictic antarctic Ekho Lake. AB - Two Gram-negative, aerobic, pointed and budding bacteria were isolated from various depths of hypersaline, heliothermal and meromictic Ekho Lake (Vestfold Hills, East Antarctica). 16S rRNA gene sequence comparisons show the isolates to be phylogenetically close to the genera Sulfitobacter and Roseobacter. Cells can be motile and contain storage granules. Sulfite addition does not stimulate growth. Isolate EL-38T can produce bacteriochlorophyll a and has a weak requirement for sodium ions; polar lipids include phosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine and an unidentified amino lipid, but not diphosphatidylgycerol. The dominant fatty acid is 18:1omega7c; other characteristic fatty acids are 3-OH 10:0, 3-OH 14:1, 16:0, 18:0, 18:2 and 19:1. The DNA base composition is 55.0-56.3 mol% G+C. Isolate EL-162T has an absolute requirement for sodium ions. Diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine and an unidentified amino lipid are present in the polar lipids. Dominant fatty acids of this isolate are 18:1omega7c and 18:1omega9c as well as 18:2 which is present as two isomers. Other characteristic fatty acids are 3-OH 10:0, 3-OH 14:1, 16:0 and 18:0. The G+C content is 57.9-58.1 mol%. Morphological, physiological and genotypic differences from related, thus far known genera support the description of Staleya guttiformis gen. nov. and sp. nov. with EL-38T (= DSM 11458T) as the type strain and of Sulfitobacter brevis sp. nov. with the type strain EL-162T (= DSM 11443T). PMID- 10826818 TI - Thermobacillus xylanilyticus gen. nov., sp. nov., a new aerobic thermophilic xylan-degrading bacterium isolated from farm soil. AB - An aerobic, thermophilic, xylanolytic, spore-forming bacterium, XETP (T = type strain; P = patent strain), has been isolated from farm soil situated underneath a manure heap in northern France. Strain XETP, which stained negative in the Gram test, occurs as short rods which sometimes form chains. Its spores are ellipsoidal, central to subterminal and occur in swollen sporangia. It grows at temperatures up to 63 degrees C and in the pH range 6.5-8.5. When grown on glucose in optimal conditions, its doubling time was found to be 33 min. CO2 was observed to have a growth-stimulating effect at the start of the culture. In addition to glucose, the isolate utilizes xylose, arabinose, mannose, cellobiose, galactose, maltose, sucrose, xylan and starch. Growth is inhibited by 5% NaCl. The G+C content of strain XETP is 57.5 mol%. The 16S rDNA sequence analysis indicated that strain XETP falls into the radiation of the Bacillus-Lactobacillus Streptococcus subdivision of the Gram-positive phylum. Its three closest phylogenetic relatives are 'Bacillus viscosus', Paenibacillus curdlanolyticus and Bacillus popilliae with identity values of 91.15, 90.94 and 90.92%, respectively. The major cellular fatty acids are 14-methyl pentadecanoic acid (16:0 iso), hexadecanoic acid (16:0) and 14-methyl hexadecanoic acid (17:0 anteiso). On the basis of 16S rRNA sequence and chemotaxonomic characteristics, the isolate is different enough for it to be considered as a member of a new genus. It is therefore proposed that this isolate represents a new genus and species: Thermobacillus xylanilyticus. Strain XETP, the type strain of Thermobacillus xylanilyticus, has been deposited in the Collection Nationale de Cultures Microbiennes (CNCM I-1017) as a patent strain. PMID- 10826819 TI - Vibrio aerogenes sp. nov., a facultatively anaerobic marine bacterium that ferments glucose with gas production. AB - A mesophilic, facultatively anaerobic, marine bacterium, designated strain FG1T, was isolated from a seagrass bed sediment sample collected from Nanwan Bay, Kenting National Park, Taiwan. Cells grown in broth cultures were motile, Gram negative rods; motility was normally achieved by two sheathed flagella at one pole of the cell. Strain FG1T required Na+ for growth, and exhibited optimal growth at 30-35 degrees C, pH 6-7 and about 4% NaCl. It grew anaerobically by fermenting glucose and other carbohydrates with production of various organic acids, including acetate, lactate, formate, malate, oxaloacetate, propionate, pyruvate and succinate, and the gases CO2 and H2. The strain did not require either vitamins or other organic growth factors for growth. Its DNA G+C content was 45.9 mol%. It contained C12:0 as the most abundant cellular fatty acid. Characterization data, together with the results of a 16S rDNA-based phylogenetic analysis, indicate that strain FG1T represents a new species of the genus Vibrio. Thus, the name Vibrio aerogenes sp. nov. is proposed for this new bacterium. The type strain is FG1T (= ATCC 700797T = CCRC 17041T). PMID- 10826820 TI - Phylogenetic analysis of Saccharothrix and related taxa: proposal for Actinosynnemataceae fam. nov. AB - Partial sequences for 16S rDNA were determined for strains of the genus Saccharothrix, including most described species, as well as strains of the described species of the related genera Kutzneria, Actinokineospora and Actinosynnema. These were aligned with published sequences for other species of these genera, as well as those of the genera 'Actinoalloteichus', 'Asiosporangium', Lentzea, Kutzneria, Streptoalloteichus and representative taxa from other actinomycete families. Phylogenetic analysis of the sequence data showed that species of the genera Actinokineospora, Actinosynnema, Lentzea and Saccharothrix are members of the same clade, and distinct from the Pseudonocardiaceae. It is proposed that a new family be created within the class Actinobacteria for these genera, to be called the Actinosynnemataceae on the basis of the oldest described genus within this family, Actinosynnema Hasegawa et al. The chemotaxonomic properties of all the genera to be placed within the new family have similar cell wall type (type III), whole-cell sugars (generally galactose although mannose and rhamnose may be present), phospholipid type (PII) and menaquinones [MK-9(H4) predominant]. PMID- 10826821 TI - Modestobacter multiseptatus gen. nov., sp. nov., a budding actinomycete from soils of the Asgard Range (Transantarctic Mountains). AB - Oligotrophic PYGV medium, inoculated with soils from Linnaeus Terrace (1600 m, Antarctica), yielded four aerobic actinomycetes with short rods, multiple and irregular septa and often motile buds. Cells were 1.0-2.8 x 1.0-3.0 microm and colonies were beige to pink. The isolates were nearly identical in physiological and biochemical tests. Three strains grew from 0 degrees C to 25-28 degrees C, but one was psychrophilic with a maximum growth temperature of 20 degrees C. Carbon sources utilized were D-glucose, D-galactose, lactose, sucrose or mannitol; malate, succinate, fumarate, pyruvate or glutarate were decarboxylated aerobically. Peptone and yeast extract were the preferred nitrogen sources. Nitrate was reduced aerobically or anaerobically. Cell walls contained meso diaminopimelic acid, glutamate, alanine, glycine, galactose, glucose and ribose. Major fatty acids of strains AA-802, -824, -825 and -826T were n18:1, i16:0 and ai17:0. Major respiratory quinones were MK-9(H4) and MK-8(H4). Polar lipids were diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylinositol. Phosphatidylglycerol was found in most strains. The DNA G+C contents were 68-70 mol%. In 16S rDNA analyses, similarity values obtained for 500 nucleotides from the 5' terminus were > 99.5%. Almost complete sequences from AA-826T and -825 were 99.9% similar. Strain AA-826T belonged to a novel cluster of desert soil and rock isolates within the Geodermatophilaceae and was equidistantly related to members of Geodermatophilus and to a Blastococcus lineage. The four isolates represent a new genus, Modestobacter gen. nov., with Modestobacter multiseptatus sp. nov. as the type species. The type strain, Modestobacter multiseptatus AA 826T, was deposited in the DSMZ as DSM 44406T. PMID- 10826822 TI - Corynebacterium simulans sp. nov., a non-lipophilic, fermentative Corynebacterium. AB - Three coryneform strains isolated from clinical samples were analysed. These strains fitted the biochemical profile of Corynebacterium striatum by conventional methods. However, according to recently described identification tests for fermenting corynebacteria, the strains behaved rather like Corynebacterium minutissimum. The three isolates could be distinguished from C. minutissimum by a positive nitrate and nitrite reductase test and by not fermenting maltose; from C. striatum by their inability to acidify ethylene glycol and to grow at 20 degrees C. Genetic studies based on 16S rRNA showed that the three strains were in fact different from C. minutissimum and C. striatum (96.9 and 98% similarity, respectively) and from other corynebacteria. They represent a new species for which the name Corynebacterium simulans sp. nov. is proposed. The type strain is DSM 44415T (= UCL 553T = Co 553T). PMID- 10826823 TI - Frigoribacterium faeni gen. nov., sp. nov., a novel psychrophilic genus of the family Microbacteriaceae. AB - The taxonomic position of five actinobacterial strains isolated from dust, an animal shed, the air inside a museum and soil was investigated using a polyphasic approach. The growth characteristics were unusual for actinomycetes. Optimal growth was at temperatures ranging from 2 to 10 degrees C. After small-step adaptation (5 degrees C steps) to higher temperatures, the strains were also able to grow at 20 degrees C. Cell wall analyses revealed that the organisms showed a hitherto undescribed, new group B-type peptidoglycan [type B2beta according to Schleifer & Kandler (1972), but with lysine instead of ornithine]. All strains contained menaquinone MK-9. Mycolic acids were not detected. Diphosphatidylglycerol, phosphatidylglycerol and an unknown glycolipid were detected in the polar lipid extracts. The main fatty acids were 12-methyl tetradecanoic acid (15:0 anteiso), 12-methyl-tetradecenoic acid (15:1 anteiso), 14-methyl-pentadecanoic acid (16:0 iso) and 14-methyl-hexadecanoic acid (17:0 iso), as well as an unusual compound identified as 1,1-dimethoxy-anteiso pentadecane (15:0 anteiso-DMA). The G+C content of DNA was approximately 71 mol%. The results of 16S rRNA gene sequence comparisons revealed that the strains represent a new lineage in the suborder Micrococcineae and the family Microbacteriaceae of the order Actinomycetales. On the basis of these results the new genus Frigoribacterium gen. nov. is proposed, harbouring the new species Frigoribacterium faeni sp. nov. (type strain = 801T = DSM 10309T). PMID- 10826824 TI - The genus Abiotrophia (Kawamura et al.) is not monophyletic: proposal of Granulicatella gen. nov., Granulicatella adiacens comb. nov., Granulicatella elegans comb. nov. and Granulicatella balaenopterae comb. nov. AB - The genus Abiotrophia currently includes four species, Abiotrophia defectiva, Abiotrophia adiacens, Abiotrophia balaenopterae and Abiotrophia elegans. Recent 16S rRNA gene sequencing studies have demonstrated that the genus is not monophyletic and is in need of taxonomic revision. Phylogenetically, the genus Abiotrophia consists of two distinct lines, A. defectiva, the type species of the genus, and a robust group consisting of A. adiacens, A. balaenopterae and A. elegans. Therefore, it is formally proposed that the genus Abiotrophia should be restricted to A. defectiva and that A. adiacens, A. balaenopterae and A. elegans should be reclassified in a new genus, Granulicatella, as Granulicatella adiacens comb. nov., Granulicatella balaenopterae comb. nov. and Granulicatella elegans comb. nov. PMID- 10826825 TI - Leifsonia poae gen. nov., sp. nov., isolated from nematode galls on Poa annua, and reclassification of 'Corynebacterium aquaticum' Leifson 1962 as Leifsonia aquatica (ex Leifson 1962) gen. nov., nom. rev., comb. nov. and Clavibacter xyli Davis et al. 1984 with two subspecies as Leifsonia xyli (Davis et al. 1984) gen. nov., comb. nov. AB - The new genus Leifsonia gen. nov. with two new species, Leifsonia poae sp. nov. (type strain VKM Ac-1401T) and Leifsonia aquatica (ex Leifson 1962) nom. rev., comb. nov. (the type species, with VKM Ac-1400T = DSM 20146T = JCM 1368T as type strain), is proposed to accommodate bacteria found in Poa annua root gall, induced by the nematode Subanguina radicicola, and 'Corynebacterium aquaticum' Leifson 1962. Further, it is proposed to reclassify Clavibacter xyli Davis et al. 1984 with two subspecies in the new genus as Leifsonia xyli (Davis et al. 1984) comb. nov., Leifsonia xyli subsp. xyli (Davis et al. 1984) comb. nov. and Leifsonia xyli subsp. cynodontis (Davis et al. 1984) comb. nov. Members of the proposed genus are characterized by coryneform morphology, peptidoglycans based upon 2,4-diaminobutyric acid, the major menaquinone MK-11, phosphatidylglycerol and diphosphatidylglycerol as principal phospholipids, the high content of anteiso- and iso-branched saturated fatty acids, and a DNA G+C base composition of 66-73 mol%. They form a distinct phylogenetic branch attached to the line of descent of Agromyces spp. The new and reclassified species of the new genus clearly differ from each other phylogenetically and phenetically and can be recognized by their morphologies, the cell wall sugar composition, the requirement of complex media for growth, and numerous physiological characteristics, including the oxidase reaction. PMID- 10826826 TI - Trichosporon veenhuisii sp. nov., an alkane-assimilating anamorphic basidiomycetous yeast. AB - A morphological and physiological description of an alkane-assimilating anamorphic basidiomycetous yeast species, named Trichosporon veenhuisii, is presented. The ability to assimilate several aliphatic and aromatic compounds as sole source of carbon and energy is reported. The phylogenetic position within the genus, based on nuclear base sequencing of the D1/D2 region of the large subunit of rDNA, is discussed. The type strain is CBS 7136T. PMID- 10826827 TI - Candida tartarivorans sp. nov., an anamorphic ascomycetous yeast with the capacity to degrade L(+)- and meso-tartaric acid. AB - An undescribed anamorphic yeast species of ascomycetous affinity, for which the name Candida tartarivorans is proposed, was isolated from dried wine lees in Portugal using a selective medium with L(+)-tartaric acid as the sole source of carbon and energy. The single isolate (IGC 4854T) showed the following characteristics: sympodial holoblastic conidiogenesis, absence of asci with ascospores, a negative colour reaction with Diazonium Blue B, production of elaborate pseudomycelium and ability to grow with inositol as sole source of carbon. Analysis of the physiological data pointed to a close relationship with other inositol-assimilating taxa, namely the genera Arxula, Stephanoascus, Sympodiomyces, Zygoascus and selected Candida species. Comparative analysis of the D1/D2 variable domain of the 26S rRNA gene of all available sequences for ascomycetous yeasts showed that strain IGC 4854T did not match with any other species in the database. The closest relative was Candida auringiensis Santa Maria, but the two species differed in 24 nucleotide positions. A description of the new species is given. PMID- 10826828 TI - Four new yeasts in the Pichia anomala clade. AB - Four new yeasts are described that were recognized as novel from nucleotide substitutions in domain D1/D2 of 26S rDNA, a region that is sufficiently divergent to allow resolution of most ascomycetous yeast species. The new species and their type strains are as follows: Pichia maclurae NRRL Y-5377T (= CBS 8671T); Pichia misumaiensis NRRL Y-17389T (= CBS 8062T); Candida mycetangii NRRL Y-6843T (= CBS 8675T); and Candida ulmi NRRL YB-2694T (= CBS 8670T). The two Pichia species form spherical ascospores and are heterothallic. Phylogenetic analysis of domain D1/D2 sequences placed the four new species in the Pichia anomala clade. PMID- 10826829 TI - Phylogeny of the genus Kluyveromyces inferred from the mitochondrial cytochrome-c oxidase II gene. AB - A phylogenetic analysis of 17 species belonging to the genus Kluyveromyces and 12 reference and outgroup species was performed using mitochondrial cytochrome-c oxidase II gene sequences. The genus Kluyveromyces appears as a polyphyletic taxon formed by species included within the following four main groups. The Kluyveromyces phaffii group encompasses the species Kluyveromyces blattae, K. phaffii and Kluyveromyces yarrowii. The Kluyveromyces marxianus group is a monophyletic group consisting of the species Kluyveromyces aestuarii, Kluyveromyces dobzhanskii, Kluyveromyces lactis, K. marxianus and Kluyveromyces wickerhamii. The monophyletic Kluyveromyces thermotolerans group is formed by K. thermotolerans, Kluyveromyces waltii and Saccharomyces kluyveri (which appears in the mitochondrial tree as the sister clade of the K. marxianus group). Finally, the Saccharomyces cerevisiae group contains the remaining Kluyveromyces species, as well as the reference Saccharomyces species (sensu lato and sensu stricto) and Candida glabrata (the phylogenetic relationships within this group are unclear according to the bootstrap test). The phylogenetic relationships obtained for this mitochondrial gene are, for the most part, congruent with previous trees based on nuclear rRNA sequences, except for the position of K. yarrowii and the close relationship between the K. marxianus and K. thermotolerans groups. These differences, as well as the existence of these groups, are discussed in the context of previous studies based on phenotypic, genetic and molecular data. Although additional studies are required to decipher the phylogenetic relationships between the genus Kluyveromyces and the closely related genera Saccharomyces, Torulaspora and Zygosaccharomyces, future changes to their taxonomic status should take account of the existence of these four groups of Kluyveromyces species. PMID- 10826830 TI - Taxonomic relationships among the taxa in the Candida guilliermondii complex, as revealed by comparative electrophoretic karyotyping. AB - Electrophoretic karyotypes of 15 type strains of the taxa in the Candida guilliermondii complex including Candida fukuyamaensis Nakase et al. and Candida xestobii Yarrow et S. A. Meyer were comparatively analysed by using the CHEF (contour-clamped homogeneous electric field) method of PFGE. Eighteen strains (isolated from various natural sources in China) which were originally identified as C. guilliermondii by conventional methods were also included. Six electrophoretic karyotype groups were recognized among the strains compared. The following type strains were grouped together with the type strains of C. guilliermondii (Castellani) Langeron et Guerra and Pichia guilliermondii Wickerham: Blastodendrion arztii Ota, Blastodendrion krausi Ota, Candida amidovorans Balloni et al., C. guilliermondii var. japonica Sugiyama et Goto, Candida mamillae S. Goto, Candida parapsilosis (Ashford) Langeron et Talice var. tokyoensis Suzuki et al., C. parapsilosis var. tuxtlensis Herrera et al. and six Chinese strains. The type strain of Torulopsis kestonii Scarr et Rose was classified into the group together with the type strain of Candida fermentati (Saito) Bai and seven Chinese strains. The group represented by the type strain of C. fukuyamaensis included five other strains isolated in China. The type strains of Candida xestobii, C. guilliermondii var. carpophila Phaff et M. W. Miller and Trichosporon appendiculare Batista et al. were separated into three different groups, respectively. Taxonomic relationships among the taxa studied are discussed. PMID- 10826831 TI - Pooled-sample testing as a herd-screening tool for detection of bovine viral diarrhea virus persistently infected cattle. AB - The study was conducted to develop methodology for least-cost strategies for using polymerase chain reaction (PCR)/probe testing of pooled blood samples to identify animals in a herd persistently infected with bovine viral diarrhea virus (BVDV). Cost was estimated for 5 protocols using Monte Carlo simulations for herd prevalences of BVDV persistent infection (BVDV-PI) ranging from 0.5% to 3%, assuming a cost for a PCR/probe test of $20. The protocol associated with the least cost per cow involved an initial testing of pools followed by repooling and testing of positive pools. For a herd prevalence of 1%, the least cost per cow was $2.64 (95% prediction interval = $1.72, $3.68), where pool sizes for the initial and repooled testing were 20 and 5 blood samples per pool, respectively. Optimization of the least cost for pooled-sample testing depended on how well a presumed prevalence of BVDV-PI approximated the true prevalence of BVDV infection in the herd. As prevalence increased beyond 3%, the least cost increased, thereby diminishing the competitive benefit of pooled testing. The protocols presented for sample pooling have general application to screening or surveillance using a sensitive diagnostic test to detect very low prevalence diseases or pathogens in flocks or herds. PMID- 10826832 TI - Type C botulism in dairy cattle from feed contaminated with a dead cat. AB - Four hundred twenty-seven of 441 adult Holstein dairy cattle from a 1,200-cow dairy died over a 1-week period during early spring 1998. Affected animals were from 4 late lactation pens, one of which included the bull string. Signs included weakness, recumbency, watery diarrhea, and death. Eighty animals from the 4 pens were dead approximately 8 hours after the first ill cows were noted. Affected cows would collapse on stimulation and extend all 4 limbs with moderate rigidity. Several lacked lingual tonus and had abdominal breathing patterns. The animals had been fed a load of total mixed ration that included a rotten bale of oat hay containing a dead cat. No common toxicants were identified, and pathologic examination revealed no consistent lesions. Testing of tissue from the cat carcass found in the feed sample using mouse protection bioassay identified the presence of type C botulinum toxin. Samples of feed, tissue from affected animals, cat tissue from feed, milk, and serum were also tested using an enzyme linked immunosorbent assay (ELISA) specific for type C botulinum. Two samples of rumen contents were tested and found to be positive for botulism by ELISA, and 1 of 3 liver samples had a weak positive finding. No botulinum toxin was found in milk or sera using the ELISA. PMID- 10826833 TI - Urinary alkaloid excretion as a diagnostic tool for fescue toxicosis in cattle. AB - Fescue toxicosis research studies have often included serum prolactin as a physiologic index of the disorder. Serum prolactin has not been used as a clinical measure of fescue toxicosis because of variation associated with sex and physiologic condition of the animal and climatic and seasonal factors. The primary excretory route of the alkaloids responsible for this toxicosis is the urine. Three pasture experiments were conducted to examine serum prolactin and urinary ergot alkaloid variability among steers continuously grazing endophyte infected (E+) or endophyte-free (E-) tall fescue and among steers that were switched from one pasture form to the other. A fourth grazing experiment was used to examine how to best to manage the steers prior to sampling for urinary ergot alkaloid excretion. Coefficients of variability for urinary alkaloid excretion were consistently lower (46-65%) than serum prolactin (64-142%). Urinary alkaloid excretion patterns changed within 12 hours following switching steers from E+ to E- pasture or vice versa, but serum prolactin was recalcitrant to change. Because it is less variable and more dynamic than serum prolactin, urinary alkaloid excretion can be used for health assessment of steers grazing E+ and E- pastures. Regression analysis established a quadratic relationship between alkaloid excretion and average daily weight gain, with a regression coefficient of 0.86. Urinary alkaloid analysis was useful in determining whether cattle were consuming toxic tall fescue. PMID- 10826834 TI - In vitro activity of difloxacin against canine bacterial isolates. AB - The in vitro activity of difloxacin against canine bacterial isolates from clinical cases was studied in the United States and The Netherlands. Minimal inhibitory concentrations (MIC), the postantibiotic effect, the effect of pH on antimicrobial activity, and the bacterial killing rate tests were determined according to standard techniques. The MICs of American and Dutch isolates agreed in general. The MICs of the American gram-negative isolates ranged from 0.06 to 2.0 microg/ml, and the MICs of the Dutch gram-negative isolates ranged from 0.016 to 8.0 microg/ml. A few European strains of Proteus mirabilis and Klebsiella pneumoniae had relatively high MICs. Bordetella bronchiseptica also was less susceptible to difloxacin. The MICs of the American gram-positive cocci ranged from 0.125 to 4.0 microg/ml, and the MICs of Dutch isolates ranged from 0.125 to 2.0 microg/ ml. Difloxacin induced a concentration-dependent postantibiotic effect that lasted 0.2-3 hours in cultures with Escherichia coli, Staphylococcus intermedius, Streptococcus canis, Proteus spp., and Klebsiella pneumoniae. There was no postantibiotic effect observed against canine Pseudomonas aeruginosa. Decreasing the pH of the medium increased the MIC of Proteus mirabilis for difloxacin. The MICs of Escherichia coli and Klebsiella pneumoniae were lowest at neutral pH and were slightly increased in acid or alkaline media. At a neutral pH, most tested bacterial species were killed at a difloxacin concentration of 4 times the MIC. Similar results were obtained when these same bacteria were tested against enrofloxacin. A Klebsiella pneumoniae strain in an acidic environment was readily killed at difloxacin or enrofloxacin MIC, but at neutral pH the drug concentration had to be raised to 4 times the MIC for a bactericidal effect. After 24 hours of incubation at pH 7.1, difloxacin and enrofloxacin had similar bactericidal activity for all bacteria tested except Staphylococcus intermedius. Against S. intermedius, difloxacin was more bactericidal than enrofloxacin. PMID- 10826835 TI - Detection of Streptococcus suis by in situ hybridization, indirect immunofluorescence, and peroxidase-antiperoxidase assays in formalin-fixed, paraffin-embedded tissue sections from pigs. AB - Streptococcus suis is an important pathogen in pigs and is considered a zoonotic agent. To aid diagnosis of infection caused by S. suis, a species-specific probe targeting 16S ribosomal RNA was designed and used for fluorescent in situ hybridization. Two additional immunohistochemical detection methods, an indirect immunofluorescence assay and a peroxidase-antiperoxidase method, using polyclonal antibodies also were developed. The specificity of the oligonucleotide probe was examined by whole-cell and dot-blot hybridization against reference strains of the 35 serotypes of S. suis and other closely related streptococci and other bacteria commonly isolated from pigs. The probe was specific for S. suis serotypes 1-31. The specificity of the polyclonal antibodies, which has previously been evaluated for use in diagnostic bacteriology for typing of serotype 2, was further evaluated in experimentally infected murine tissue with pure culture of different serotypes of S. suis, related streptococci, and other bacteria commonly found in pigs. The polyclonal antibodies against S. suis serotype 2 cross-reacted with serotypes 1 and 1/2 in these assays. The in situ hybridization and the immunohistochemical methods were used for detection of S. suis in formalin-fixed, paraffin-embedded tissue sections of brain, endocardium, and lung from pigs infected with S. suis. The methods developed were able to detect single cells of S. suis in situ in the respective samples, whereas no signal was observed from control tissue sections that contained organisms other than S. suis. These techniques are suitable for determining the in vivo localization of S. suis for research and diagnostic purposes. PMID- 10826836 TI - Intestinal lesions caused by a strain of Chlamydia suis in weanling pigs infected at 21 days of age. AB - The objective of this study was to determine whether a strain of Chlamydia suis shown previously to be an intestinal pathogen in gnotobiotic piglets could cause diarrhea and intestinal lesions in young weanling pigs. Pigs from 2 sows were randomly assigned to 2 groups. Group 1 included 13 pigs that were weaned at 24 hours of age and then housed in isolator units and fed milk replacer and unmedicated starter ration. Group 2 included 8 pigs that nursed their respective sows, consumed unmedicated starter ration, and were weaned at 21 days of age. Ten pigs in group 1 and 6 pigs in group 2 were inoculated orally with 4 x 108 inclusion-forming units of C. suis strain R27 at 21 days of age. Control pigs were inoculated with sham inoculum. The pigs were necropsied 5-14 days postinoculation (DPI). None of the Chlamydia-infected pigs developed diarrhea. Villus atrophy was seen histologically in sections of ileum from Chlamydia infected pigs in both groups 5 and 7 days DPI. Lymphangitis and multiple lymphohistiocytic and neutrophilic aggregates were seen in the submucosa, tunica muscularis, and serosa of the distal jejunum, ileum, and colon from Chlamydia infected pigs in both groups 5-14 DPI. Immunostaining of sections of distal jejunum, ileum, and colon from infected pigs revealed chlamydial antigen in intestinal epithelium and in foci of lymphangitis/inflammation. The results indicated that C. suis strain R27 can cause intestinal lesions in young weanling pigs, and the lesions are similar to those seen in gnotobiotic piglets. The results also indicated that strain R27 causes asymptomatic intestinal infections in young weanling pigs, at least under the conditions of this study. PMID- 10826837 TI - Comparison of a clinic-based ELISA test kit with the immunofluorescence test for the assay of Ehrlichia canis antibodies in dogs. AB - The "gold standard" for the detection of antibodies to Ehrlichia canis, the cause of canine monocytic ehrlichiosis (CME), is the indirect immunofluorescence antibody (IFA) test. The IFA test however is generally available only in selected laboratories and requires extensive equipment and trained personnel. A double blind study was conducted to compare the ability of an in-clinic standardized enzyme-linked immunosorbent assay (ELISA) test kit to measure E. canis IgG antibodies in dogs compared with the standard IFA technique. A good correlation was found between the 2 techniques (r2 = 0.8793; P < 0.0001). Evidence for the sensitivity of the ELISA technique for the early detection of E. canis IgG antibodies was demonstrated by comparing the appearance of E. canis antibody titers by the IFA and ELISA techniques after artificial infection of 2 sets of dogs. In both experimental infections, both tests were equally sensitive for the early detection of IgG antibodies against E. canis, and the results correlated well with the appearance of fever and clinical signs. Proposed application of the in-clinic ELISA test is to aid in the diagnosis of CME. PMID- 10826838 TI - Single-dilution enzyme-linked immunosorbent assay for quantification of antigen specific salmonid antibody. AB - An enzyme-linked immunosorbent assay (ELISA) was developed on the basis of testing a single dilution of serum to quantify the level of antibody to the p57 protein of Renibacterium salmoninarum in sockeye salmon (Oncorhynchus nerka). The levels of antibody were interpolated from a standard curve constructed by relating the optical densities (OD) produced by several dilutions of a high-titer rainbow trout (O. mykiss) antiserum to the p57 protein. The ELISA OD values produced by as many as 36 test sera on each microplate were compared with the standard curve to calculate the antigen-specific antibody activity. Repeated measurements of 36 samples on 3 microplates on each of 6 assay dates indicated that the mean intraassay coefficient of variation (CV) was 6.68% (range, 0-23%) and the mean interassay CV was 8.29% (range, 4-16%). The antibody levels determined for the serum sample from 24 sockeye salmon vaccinated with a recombinant p57 protein generally were correlated with the levels determined by endpoint titration (r2 = 0.936) and with results from another ELISA that was based on extrapolation of antibody levels from a standard curve (r2 = 0.956). The single-dilution antibody ELISA described here increases the number of samples that can be tested on each microplate compared with immunoassays based on analysis of several dilutions of each test serum. It includes controls for interassay standardization and can be used to test fish weighing <3 g. PMID- 10826839 TI - Toxigenic Corynebacterium diphtheriae associated with an equine wound infection. PMID- 10826840 TI - Assessment of the long-term shedding pattern of Salmonella serovar choleraesuis following experimental infection of neonatal piglets. AB - In the United States, swine salmonellosis is most often attributed to infections by Salmonella serovar choleraesuis. As a host-adapted pathogen rarely found in nonswine sources, S. choleraesuis is thought to be spread primarily via horizontal transmission, with carrier animals playing an important role. Little has been reported regarding infection of neonatal piglets, particularly regarding their potential to become carriers. Evidence reported herein demonstrates that piglets experimentally infected by S. choleraesuis at 2 days of age were capable of shedding the pathogen for up to 85 days postinfection, at which time the study was concluded. This study also presents findings supporting the use of GN-Hajna as a preenrichment medium for the isolation of S. choleraesuis. PMID- 10826841 TI - Susceptibility of two bighorn sheep cell cultures to selected common ruminant viruses. PMID- 10826842 TI - A case report: evidence for type 2 bovine viral diarrhea virus (BVDV)-associated disease in beef herds vaccinated with a modified-live type 1 BVDV vaccine. PMID- 10826843 TI - Evaluation of a blocking ELISA using a urease conjugate for the detection of antibodies to pseudorabies virus. AB - A blocking enzyme-linked immunosorbent assay (ELISA) using a urease conjugate (U B-ELISA) was evaluated for screening sera for antibodies to pseudorabies virus under field conditions. A total of 764 serum samples were analyzed by U-B-ELISA. Of these, 264 were evaluated by both virus neutralization and U-B-ELISA, and the results were compared. U-B-ELISA showed 98.5% and 98.9% sensitivity and specificity, respectively. This test combines the sensitivity and specificity of the blocking ELISA format while allowing visual assessment of results. PMID- 10826844 TI - Suspected citrus pulp toxicosis in dairy cattle. AB - Thirteen lactating dairy cows from a herd of 650 died over a 6-week period. Most animals were down in milk production at 1 milking and were found dead at the next milking. Two cows had elevated heart rate and enlarged mandibular lymph nodes. Two others had azotemia, elevated heart rate, hyperglycemia, and weight loss. Necropsy of 10 cows revealed hemorrhages on the intestinal serosa and epicardium, lymphadenopathy, interstitial nephritis, small intestinal hemorrhage, and interstitial pneumonia. Histopathology showed lymphocytic to lymphogranulomatous inflammation in the heart, spleen, kidney, lymph nodes, liver, lung, pancreas, and adrenal gland. Phlebitis was present in 2 livers. The lesions resembled those of hairy vetch toxicosis, but no vetch was being fed. Similar lesions have been reported with the feeding of citrus pulp. Citrus pulp was being fed to the lactating cows and had been added to the diet 6 weeks before the first death. The syndrome resolved with elimination of citrus pulp from the diet. PMID- 10826845 TI - Acute pit gas (hydrogen sulfide) poisoning in confinement cattle. AB - Rapid deaths in confinement cattle caused by exposure to hydrogen sulfide (H2S) gas from manure pits has not been reported in the USA. In 1997, 158 cattle in 2 confinement pens were exposed to H2S gas as the manure in the pits under a slatted floor was agitated prior to pumping. Approximately 35 of the cattle were lying on the floor when the upper agitator was turned on. Within 5 minutes, many these cattle were down on their sides and paddling. Of these, 26 died within a few minutes. The survivors were treated and sent to slaughter. Cattle that did not show immediate signs of toxicosis remained clinically unaffected. Two steers that were near death were brought to the Purdue Animal Disease Diagnostic Laboratory for clinical evaluation, euthanasia, and necropsy. They were recumbent and unresponsive to visual and auditory stimuli. Necropsy examination yielded no significant gross lesions. No evidence of viral or bacterial infection was found. Ocular fluid nitrate concentrations were within normal limits, and no lead was detected in either animal. Microscopic examination revealed lesions consistent with H2S-induced central nervous system anoxia. Histologically, sections of brain demonstrated massive, diffuse cerebral cortical laminar necrosis and edema. Portions of the outer lamina contained hypereosinophilic and shrunken neurons. The subcortical white matter was vacuolated in some areas. The history, clinical signs, and histologic lesion of cerebral laminar necrosis led to a diagnosis of H2S toxicosis in these cattle. PMID- 10826846 TI - Endometrial carcinosarcoma in a sow: an immunohistochemical study. PMID- 10826847 TI - Spinal ataxia in a horse caused by an arachnoid diverticulum (cyst). PMID- 10826848 TI - Extramedullary plasmacytoma in a horse with ptyalism and dysphagia. AB - A Clydesdale mare was examined for weight loss, inappetence, ptyalism, and dysphagia. The main abnormality revealed by serum biochemistry was a marked hyperglobulinemia, and protein electrophoresis revealed a monoclonal gammopathy in the gamma region. The urine was positive for Bence Jones proteins. These findings suggested a plasma cell tumor. The neoplasm could not be located with extensive antemortem examination. At postmortem, neoplastic cells morphologically compatible with plasma cells and positive for equine IgG with imunoperoxidase staining infiltrated the pericardium, mediastinal stromal tissues, adrenal glands, meninges, atrioventricular valves, aorta, abdominal and thoracic fat, and nerves, including the trigeminal nerve. The neoplastic cells invading the cranial nerves were responsible for many of the presenting signs. PMID- 10826849 TI - Primary hepatic hemangiosarcoma with pulmonary metastasis in a New Zealand White rabbit. PMID- 10826850 TI - Papillomatous pastern dermatitis with spirochetes and Pelodera strongyloides in a Tennessee Walking Horse. AB - Papillomatous digital dermatitis is a common disease in cattle. The pastern dermatitis observed in a horse shared many of the gross characteristics of papillomatous digital dermatitis in cattle. Lesions included a mixture of proliferative and erosive changes, with a verrucose appearance in some areas. Microscopic similarities included pseudoepitheliomatous and papillomatous epidermal hyperplasia with hyperkeratosis, spongiosis of the epidermis, and intraepidermal spirochetes. The horse was also concurrently infected with Pelodera strongyloides. Papillomatous digital dermatitis in cattle is associated with poor husbandry practices. The environment of the affected horse was heavily contaminated with urine, manure, and other organic debris. Verrucous pododermatitis of horses may be the same as or similar to bovine papillomatous digital dermatitis, and these conditions have similar etiologies. PMID- 10826851 TI - Ischemic preconditioning: the endogenous power--a review of the literature. AB - With the re-emergence of surgery on a beating heart, it is important to examine the unique phenomenon of ischemic preconditioning (IPC) and its role as a form of myocardial protection under these circumstances. Ischemic preconditioning is defined as a short period of ischemia, regional or global, imposed before a more prolonged period of ischemia, which increases the myocardium's tolerance to ischemia by endogenous adaptive mechanism. Until recently, it was generally held that repeated ischemic episodes would have a cumulative adverse effect and could lead to a "stuttering infarction." PMID- 10826852 TI - Alterations of venous drug reactivity in humans: acquired and genetic factors. AB - Until recently, studies dealing with veins have almost always been the neglected part of vascular research. Recent data show an increasing rate of venous disease, and increasing evidence supports a role for veins in systemic diseases. The authors discuss and comment on findings of recent studies on venous drug reactivity. Alterations in venous reactivity to alpha- and beta-adrenergic, NO dependent, and other drugs have been shown in many genetically determined and acquired conditions, such as hypertension, smoking, and aging. In some of them, the changes of venous responsiveness are most likely secondary to another process, while in others the they seem to play a primary role in the development of systemic disease states. Studying the drug reactivity of the venous system more extensively provides useful information for clinicians and researchers and will no doubt help to further knowledge of the normal and pathologic processes of the vasculature. PMID- 10826853 TI - Prevention of coronary arterial renarrowing by continuous 3-day infusion of tissue plasminogen activator after successful percutaneous transluminal coronary angioplasty: a randomized, double-blind, placebo-controlled study. AB - Vascular injury resulting from percutaneous transluminal coronary angioplasty (PTCA) and subsequent thrombus formation are important factors in the pathogenesis of coronary arterial restenosis. The present study was undertaken to determine whether infusions of tissue plasminogen activator (tPA), beginning immediately after PTCA, would decrease the renarrowing of the coronary arterial lumen. Patients were randomized and divided into two groups: one group (15 patients, 16 lesions) received continuous, 3-day, intravenous infusions of tPA (0.25 mg/kg/day in 0.9% saline), beginning immediately after successful PTCA, while the other group (17 patients, 17 lesions) received saline alone. Coronary angiography was performed before, immediately after, 24 hours after, and 3 months after PTCA. Coronary stenosis (%), reference diameters, and minimum luminal diameters were measured by quantitative coronary angiography. The incidence of restenosis tended to be lower in the tPA group than in the placebo group (tPA group, 13%; placebo group, 41%; NS), although diameter stenoses before and immediately after PTCA were not significantly different in the two groups. Three months later, however, diameter stenoses were significantly smaller in the tPA group (35.6 +/- 13.3 vs 47.7 +/- 18.9%; P<0.05). Thus, intravenous tPA infusion beginning immediately after successful PTCA may inhibit renarrowing of the coronary arterial lumen. PMID- 10826854 TI - Retrospective analysis of Rt-pa thrombolysis combined with PGE1 in patients with peripheral arterial occlusions. AB - Although thrombolysis has been established for recanalization of acute and in part chronic peripheral artery occlusions, only smaller studies exist regarding the use of long-term rt-pa infiltration-thrombolysis. The objective of this study was to evaluate the benefit of additional long-term thrombolysis in patients with peripheral arterial occlusions for whom acute thrombolysis failed. From 1992 to 1997, 323 patients with peripheral arterial occlusions were treated with rt-pa (recombinant tissue-type plasminogen activator). When the thrombolysis failed during the first 3 hours, the thrombolytic therapy was continued as a long-term treatment with 3 mg rt-pa alternated by PGE1 (2.1 mL/hr for 3 hours, concentration: 20 microg/50 mL NaCl) every 3 hours. Additional heparin was applied in doses of 15,000 IE/24 hr or more to slightly increase the partial thromboplastin time. Angiographic controls were performed every 24 hours. If necessary, a final angioplasty was performed. In 142 of the 323 patients the occlusions were recanalized during the first 3 hours; 119 patients were treated with a long-term thrombolysis and in 72 (61%) a recanalization was ultimately achieved. Thus, the recanalization rate increased to 214 of 323 patients (p < 0.02). Mean treatment time was 2.8 +/- 2.2 days, range: 1 to 13 days. The rates of recanalization were not different in iliac, femoral, or crural arteries. Fibrinogen levels did not decrease during thrombolysis. Severe bleeding (with a decrease of more than 3 g/dL hemoglobin requiring transfusion) occurred in four patients after finishing the thrombolysis with short-term and in six with long term therapy; two required surgical treatment. The 1-, 2-, and 3-year cumulative patency rates were respectively 90.1%, 74.2% and 64.9%. Patency rates in patients with acute or long-term thrombolysis were not different. A composite thrombolytic treatment using low-dose rt-pa in combination with PGE1 offers significantly better results than an acute thrombolytic treatment alone. It can be an effective and practicable regimen in about 60% of patients in whom acute thrombolysis fails. PMID- 10826855 TI - Circulating biochemical marker levels of collagen metabolism are abnormal in patients with abdominal aortic aneurysm. AB - Changes in extracellular matrix composition induced by abnormal collagen metabolism in the aortic wall may be an important factor in the progression of aortic structural changes. The authors have measured several types of biochemical marker for collagen metabolism in plasma: carboxyterminal propeptide of type Icollagen (PICP) for a pure collagen synthesis marker, matrix metalloproteinase-1 (MMP-1) for a degradation marker of collagen matrix, and tissue inhibitors of metalloproteinase-1 (TIMP-1) as a native inhibitor of MMP-1. Subjects of this study were 17 patients with abdominal aortic aneurysm (AAA), 14 patients with atherosclerosis obliterans (ASO), and 22 age/sex-matched healthy controls (HC). Blood samples were drawn from a forearm vein and measured by radioimmunoassay or enzyme-linked immunosorbent assay. Plasma concentrations of PICP in patients with AAA were significantly decreased compared to those in HC patients (82.0 +/- 16.4 vs 111.3 +/- 40.3 ng/mL; p < 0.01), but those in patients with ASO (105.4 +/- 55.4 ng/mL) were comparable to control concentrations. Although no differences in plasma concentrations of MMP-1 were observed among the three subject groups (HC, 20.0 +/- 5.6 ng/mL; ASO, 21.4 +/- 13.8 ng/mL; AAA, 24.5 +/- 11.7 ng/mL; NS), MMP 1/PICP ratio as an index of collagen degradation to collagen neosynthesis in AAA was significantly elevated compared to HC (0.32 +/- 0.18 vs 0.20 +/- 0.08; p < 0.01). Plasma concentrations of TIMP-1 in patients with AAA (293.8 +/- 61.2 ng/mL) or ASO (327.6 +/- 54.9 ng/mL) were significantly higher than in HC (227.3 +/- 60.2 ng/mL; both p < 0.01). In conclusion, these data suggest that although a compensatory mechanism such as increased TIMP-1 may be activated, collagen neosynthesis may decrease with relatively increased collagen degradation in patients with AAA. PMID- 10826856 TI - Impact of a clinical pathway on the care and costs of myocardial infarction. AB - A time series of 110 patients with acute myocardial infarction admitted between January 1992 and June 1997 examined the effects of a clinical pathway. The pathway reduced length of hospital stay by 2.2 days and hospital charges by $1,008 without compromising care quality and outcomes. PMID- 10826857 TI - Effect of beta-blocker on metabolism and contraction of doxorubicin-induced cardiotoxicity in the isolated perfused rabbit heart. AB - The effect of beta-blocker (propranolol) on the metabolism and contraction of doxorubicin-induced cardiomyopathy during pacing or ischemia was examined by the phosphorus 31-nuclear magnetic resonance (31 P-NMR) in Langendorff hearts of chronically treated rabbits after cumulative doses of 16 mg doxorubicin/kg. After 8 weeks of doxorubicin treatment, beta-blocker (propranolol) was given orally over a period of 2 weeks for a cumulative dose of 1.4 mg/kg. Isolated hearts were paced at higher heart rates, or hearts were perfused on low flow. Adenosine triphosphate (ATP), creatine phosphate (PCr), inorganic phosphate (Pi), pH, left ventricular systolic developed pressure (LVDev P), and coronary flow were measured. The hearts were divided into three experimental groups: Group I consisted of controls, Group II consisted of doxorubicin treatment, and Group III consisted of doxorubicin treatment with propranolol. Group II showed a significant decrease of ATP during pacing (48 +/- 2%) and during low flow (61 +/- 6%) compared with Group I (86 +/- 9% at pacing, 94 +/- 6% on low flow). But Group III showed a significantly marked improvement of ATP during pacing (95 +/- 10%) and during low flow (83 +/- 3%) compared with Group II. Furthermore, Group II showed a significant decrease of LVDev P during pacing (69 +/- 6 mm Hg) and during low flow (63 +/- 3 mm Hg) compared with Group I (101 +/- 5 mm Hg at pacing, 95 +/- 9 mm Hg on low flow). But Group III showed a significantly marked improvement of LVDev P during pacing (93 +/- 5 mm Hg) and during low flow (83 +/- 14 mm Hg) compared with Group II. In conclusion, propranolol had a significant beneficial effect on metabolism and contraction during high-energy demand and during low oxygen supply of doxorubicin cardiomyopathy. PMID- 10826858 TI - Percutaneous coronary intervention as a bridge to renal transplantation in a patient with end-stage renal disease--a case report. AB - Renal transplantation is one of the preferred modes of replacement therapy in patients with end-stage renal disease. Cardiovascular disease remains the leading cause of morbidity and mortality in patients with end-stage renal disease and renal transplant recipients. The authors describe a patient with end-stage renal disease who developed unstable angina before renal transplantation. Emergent cardiac catheterization and percutaneous coronary intervention served as a bridge to his successful renal transplantation without complications. PMID- 10826859 TI - Prothrombin gene 20210 G-->A and Factor V Arg 506 to Gln mutation in a patient with Buerger's disease--a case report. AB - Thromboangiitis obliterans, or Buerger's disease, is a segmental occlusive inflammatory disorder of the arteries and veins most commonly affecting the lower extremities of young male cigarette smokers. The etiopathogenesis of the thromboangiitis obliterans is still obscure. The authors have identified heterozygosity for the recently described prothrombin gene 20210 G-->A variation and Factor V Arg 506 to Gln (Factor V Leiden) mutation in a patient with Buerger's disease. Both mutations confer a high risk of thrombosis. This coincidental observation may serve as further evidence that a thrombotic mechanism is involved in Buerger's disease. PMID- 10826860 TI - Inappropriate shocks diagnosed by stored electrograms of implantable cardioverter defibrillators--two case reports. AB - An implantable cardioverter defibrillator is an important therapeutic option for patients with high risk of life-threatening ventricular arrhythmias. However, their use is also associated with several complications including inappropriate shock. Although the most frequent cause of inappropriate shock is supraventricular tachyarrhythmias, lead fracture can also be associated with inappropriate shock. Diagnosis of lead fracture can be made by chest x-ray radiography, fluoroscopic examination, interrogation of the device, and intracardiac electrograms. In this report, the authors present two cases of inappropriate shock due to lead fractures in the costoclavicular region that could only be diagnosed by the help of stored intracardiac electrograms. Methods for diagnosis of lead fractures and modalities to avoid recurrences are also discussed. PMID- 10826861 TI - Monoclonal cryoglobulinemia with extensive gangrene of all four extremities--a case report. AB - The monoclonal immunoglobulin products of plasma cell neoplasm can give rise to a variety of manifestations including hyperviscosity, amyloidosis, cryoglobulinemia, neuropathy, and renal failure. A 50-year-old woman with monoclonal cryoglobulinemia developed extensive gangrene and ulceration of both hands and feet over several weeks requiring bilateral below-elbow and below-knee amputations. This case illustrates the importance of qualitative properties of paraprotein. PMID- 10826862 TI - Takayasu's arteritis concurrent with Marfan syndrome--a case report. AB - Marfan syndrome (MS) is a dominantly inherited connective tissue disorder characterized by arachnodactyly, tall stature, the presence of aortic aneurysm, and lens dislocation. Takayasu's arteritis (TA) is a chronic vasculitis that primarily affects the aorta and its branches. The authors report the first case of TA in a patient with MS. The simultaneous presence of TA and MS could be a coincidence, however; the pathogenesis of TA might be linked with autoimmunity induced by abnormal extracellular matrix protein derived from the genetic mutations in MS. PMID- 10826863 TI - Obesity management in primary care: assessment of readiness to change among 284 family practice patients. AB - BACKGROUND: Most adults in primary care are overweight or obese; two thirds of patients with weight problems have other obesity-related conditions. The study objective was to explore the feasibility of a primary care obesity intervention based on the transtheoretical model (TM) of behavior change and principles of chronic disease (CD) care. METHODS: A prospective study of the initial version of the TM-CD intervention with obese family practice patients (n = 284) yielded cross-sectional data on baseline stage of change for six target behaviors: dietary fat, portion control, vegetable intake, fruit intake, usual physical activity, and planned exercise. The sample consisted of obese patients scheduled for an office visit during times when recruitment and informed consent did not conflict with acute care. RESULTS: Obese patients volunteering for a TM-CD program are in different stages of change for six target behaviors. Preparation was the most frequently reported stage for increased exercise (49%) or activity (34%), decreased dietary fat consumption (44%), and increased portion control (51%). Patients in a particular stage for one behavior were distributed across all five stages for another behavior. Stage of change for five target behaviors was associated with body mass index or waist girth (P < .05) in a manner consistent with stage-of-change theory. CONCLUSIONS: Using the transtheoretical model of behavior change will allow physicians to recognize when obese patients are receptive to specific behavioral interventions. PMID- 10826864 TI - Are yearly physical examinations in adolescents necessary? AB - BACKGROUND: Recommendations regarding the frequency of routine physical examinations for adolescents have varied from one examination every 2 to 3 years to yearly evaluations. Because none of these recommendations was based on studies regarding the usefulness of such examinations, it was pertinent to review the results of published studies. METHODS: All series of routine school and preathletic examinations of adolescents published in the English literature from 1943 to 1995 were reviewed. Only reviews of examinations by physicians with or without supervised health professionals were included. RESULTS: Findings included weight, blood pressure, visual acuity, innocent heart murmurs, scoliosis, referral for further testing, and serious abnormalities unknown before examination. A total of 20,047 examinations by 12 different groups of investigators was abstracted. Only 2 adolescents had major, previously unknown findings: 1 boy was blind in one eye and the other had mitral insufficiency. Elevated blood pressures were found in 0.1% to 1.6% of adolescents. Minor findings included acne, caries, myopia, and minor orthopedic problems, but they did not prevent participation in school or sports. CONCLUSIONS: Yearly physical examinations in adolescents are not cost-effective and have practically no value in finding important pathologic conditions. This conclusion would not apply to sexually active teenagers. The value of an examination for health education or detection of mental problems has never been tested in this population. For entrance to school and camps or for sports participation, the review of a questionnaire and screening examinations by allied health providers should be the method of choice unless future studies justify repeated yearly examination of adolescents. PMID- 10826865 TI - Impact of vaginal antifungal products on utilization of health care services: evidence from physician visits. AB - BACKGROUND: Recently many medications formerly available only by prescription have been approved for over-the-counter (OTC) status. In 1990, clotrimazole became the first available OTC drug to treat candidal vaginitis. Subsequently several other prescription antifungal medications have also been available in OTC products. One proposed benefit of these switches from prescription to OTC status is a reduction in the utilization of health care services. METHODS: Using National Ambulatory Medical Care Survey data, the average numbers of annual visits for vaginal complaints were estimated for 1985, 1990, and 1994. These years were chosen because they represented periods before, close to, and after the approval of the OTC antifungal preparations. The estimated visits for each year were compared using a chi-square analysis with a sample weight correction. RESULTS: There was a 15% decline in the number of vaginitis visits from 1990 to 1994 that potentially could be attributed to the availability of the OTC antifungal preparations. The decrease in physician visits results in approximately $45 million in direct cost savings and another $18.75 million in indirect savings by reducing time lost from work. CONCLUSIONS: It appears that the availability of OTC anticandidal fungal preparations reduces the number of physician visits for vaginitis, resulting in cost savings. PMID- 10826866 TI - Perceptions of practice among rural family physicians--is there a gender difference? AB - BACKGROUND: Women physicians are relatively less likely to practice in rural areas. The purpose of this study was to describe differences in perceptions of rural practice between male and female physicians currently in rural practice. METHODS: A questionnaire was developed from interviews with rural family physicians and mailed to 118 randomly selected board-certified family physicians in rural towns of the northwestern United States. RESULTS: Responses from 63 physicians were analyzed quantitatively and qualitatively. Female family physicians were more likely than their male counterparts to agree they had been attracted by the availability of part-time work and by opportunities for their personal partner. Compared with men, women were more likely to expect to earn less in rural than in urban practice. On open-ended questions analyzed using qualitative methods, women were more likely to report discouragement by professional isolation and by potential lack of privacy in rural areas. CONCLUSIONS: These responses suggest that during recruitment efforts rural communities might be able to attract more female physicians by offering flexible schedules, spouse or partner opportunities, role boundaries, and professional or personal support networks. PMID- 10826867 TI - Primum non nocere and the quality of evidence: rethinking the ethics of screening. AB - BACKGROUND: Screening is different from investigation, and these differences have important implications in the assessment of screening programs. METHODS: I review the differences between screening and investigation and the implications of these differences derived from a consideration of the four ethical principles of beneficence, nonmaleficence, autonomy, and distributive justice. RESULTS: Because most of the harms of screening fall on the healthy and because screening is initiated by physicians, nonmaleficence takes ethical precedence over beneficence. Issues related to cost and consent are also approached differently in screening compared with investigation, and both take on greater ethical importance. I contend further that these ethical implications require that screening programs be backed up by better evidence than is the usual case for investigative medicine. I suggest an outline for the appropriate assessment of screening programs and for the ethical responsibilities of those involved in screening. CONCLUSIONS: Many current medical screening practices are not concordant with our ethical principles and should be reassessed. PMID- 10826868 TI - A case of fatal food-borne septicemia: can family physicians provide prevention? AB - BACKGROUND: Vibrio vulnificus, a common bacteria found in undercooked seafood and seawater, is the leading cause of food-borne death in Florida. Fatal cases of V vulnificus infection have also been reported in most states. METHODS: The literature was searched using the key words "Vibrio vulnificus," "septicemia," "wound infections," "seafood," "immunocompromise," and "patient education." A case of fatal V vulnificus septicemia is described. RESULTS AND CONCLUSIONS: V vulnificus, part of the natural flora of temperate coastal waters and one of the most abundant microorganisms found in seawater, has been isolated from waters off the Gulf, Pacific, and Atlantic coasts of the United States. Infections in noncoastal regions have been traced to consumption of seafood derived from Gulf Coast waters. Seawater exposure and consumption of inadequately cooked seafood are routes most commonly associated with V vulnificus infection. Exposure to V vulnificus is life-threatening for chronically ill or immunocompromised patients, who are most likely to develop fatal septicemia. Currently a combination of doxycycline and intravenous ceftazidime is recommended treatment. Mortality rates from V vulnificus continue to be high in immunocompromised patients. Family physicians can help prevent this outcome by counseling high-risk patients. PMID- 10826869 TI - Applying a meta-analysis to daily clinical practice. PMID- 10826870 TI - Trimethoprim-sulfamethoxazole-induced hypoglycemia as a cause of altered mental status in an elderly patient. PMID- 10826871 TI - Herpetic whitlow of the toe. PMID- 10826872 TI - Cervical prolapse during pregnancy. PMID- 10826873 TI - Cushing syndrome and adrenocortical carcinoma in a patient with CD4+ lymphocytopenia. PMID- 10826874 TI - The challenge of obesity management in primary care. PMID- 10826875 TI - Annual physical examination for adolescents: a reassessment. PMID- 10826876 TI - Teaching information mastery. PMID- 10826877 TI - Congestive heart failure clinical outcomes study in a private community medical group. PMID- 10826878 TI - Prenatal testing and counseling for Down syndrome. PMID- 10826879 TI - Delivery of preventive services. PMID- 10826880 TI - Characterization of the hydrogen bond network in guanosine quartets by internucleotide 3hJ(NC)' and 2hJ(NN) scalar couplings. AB - Scalar coupling correlations across hydrogen bonds with carbonyl groups as acceptors have been observed in a variety of proteins, but not in nucleic acids. Here we present a pulse scheme that allows such an observation and quantification of trans-hydrogen bond 3hJ(NC)' correlations in nucleic acid base pairs, between the imino nitrogen 15N1 and the carbonyl 13C6 nuclei within the guanine quartets of the Oxy-1.5 DNA-quadruplex. Intra- and internucleotide N-H...O=C connectivities can be traced around each guanine quartet, allowing the hydrogen bonding partners to be unambiguously assigned. Absolute values of the 3hJ(NC)' couplings are approximately 0.2 Hz as quantified by a selective long-range H(N)CO experiment and are thus on average smaller than the analogous 3hJ(NC)' couplings observed in proteins. In addition, an improved version of the pseudo heteronuclear H(N)N-COSY [Majumdar et al. (1999) J. Biomol. NMR, 14, 67-70] is presented which allows simultaneous detection of the 15N-donor and 15N-acceptor resonances connected by 2hJ(NN) couplings in hydrogen bonds involving amino groups. Using this experiment, values ranging between 6 and 8 Hz are determined for the 2hJNN couplings between 15N2 and 15N7 nuclei in the guanine quartet. These values are not strongly influenced by the presence of a significant amount of chemical exchange broadening due to amino group rotations. PMID- 10826881 TI - Transverse relaxation optimized triple-resonance NMR experiments for nucleic acids. AB - Triple resonance HCN and HCNCH experiments are reliable methods of establishing sugar-to-base connectivity in the NMR spectra of isotopicaly labeled oligonucleotides. However, with larger molecules the sensitivity of the experiments is drastically reduced due to relaxation processes. Since the polarization transfer between 13C and 15N nuclei relies on rather small heteronuclear coupling constants (11-12 Hz), the long evolution periods (up to 30 40 ms) in the pulse sequences cannot be avoided. Therefore any effort to enhance sensitivity has to concentrate on manipulating the spin system in such a way that the spin-spin relaxation rates would be minimized. In the present paper we analyze the efficiency of the two known approaches of relaxation rate control, namely the use of multiple-quantum coherence (MQ) and of the relaxation interference between chemical shift anisotropy and dipolar relaxation - TROSY. Both theoretical calculations and experimental results suggest that for the sugar moiety (H1'-C1'-N1/9) the MQ approach is clearly preferable. For the base moiety (H6/8-C6/8-N1/9), however, the TROSY shows results superior to the MQ suppression of the dipole-dipole relaxation at moderate magnetic fields (500 MHz) and the sensitivity improvement becomes dramatically more pronounced at very high fields (800 MHz). The pulse schemes of the triple-resonance HCN experiments with sensitivity optimized performance for unambiguous assignments of intra-residual sugar-to-base connectivities combining both approaches are presented. PMID- 10826882 TI - Solution structure of the human BTK SH3 domain complexed with a proline-rich peptide from p120cbl. AB - X-linked agammaglobulinemia (XLA), an inherited disease, is caused by mutations in the Bruton's tyrosine kinase (BTK). The absence of functional BTK leads to failure of B cell differentiation which incapacitates antibody production in XLA patients leading to, sometimes lethal, bacterial infections. Point mutation in the BTK gene that leads to deletion of C-terminal 14 aa residues of BTK SH3 domain was found in one patient family. To understand the role of BTK in B cell development, we have determined the solution structure of BTK SH3 domain complexed with a proline-rich peptide from the protein product of c-cbl protooncogene (p120cbl). Like other SH3 domains, BTK SH3 domain consists of five beta-strands packed in two beta-sheets forming a beta-barrel-like structure. The rmsd calculated from the averaged coordinates for the BTK SH3 domain residues 218 271 and the p120cbl peptide residues 6-12 of the complex was 0.87 A (+/-0.16 A) for the backbone heavy atoms (N, C, and Calpha) and 1.64 A (+/-0.16 A) for all heavy atoms. Based on chemical shift changes and inter-molecular NOEs, we have found that the residues located in the RT loop, n-Src loop and helix-like loop between beta4 and beta5 of BTK SH3 domain are involved in ligand binding. We have also determined that the proline-rich peptide from p120cbl binds to BTK SH3 domain in a class I orientation. These results correlate well with our earlier observation that the truncated BTK SH3 domain (deletion of beta4, beta5 and the helix-like loop) exhibits weaker affinity for the p120cbl peptide. It is likely that the truncated SH3 domain fails to present to the ligand the crucial residues in the correct context and hence the weaker binding. These results delineate the importance of the C-terminus in the binding of SH3 domains and also indicate that improper folding and the altered binding behavior of mutant BTK SH3 domain likely lead to XLA. PMID- 10826883 TI - Probing hydrogen bonds in the antibody-bound HIV-1 gp120 V3 loop by solid state NMR REDOR measurements. AB - We describe solid state NMR measurements on frozen solutions of the complex of the 24-residue HIV-1 gp120 V3 loop peptide RP135 with the Fab fragment of the anti-gp120 antibody 0.5beta, using rotational echo double resonance (REDOR). In order to probe possible hydrogen bonding between arginine side chains and glycine backbone carbonyls in the region of the conserved Gly-Pro-Gly-Arg (GPGR) motif of the V3 loop, RP135 samples were prepared with 15N labels at the eta nitrogen positions of arginine side chains and 13C labels at glycine carbonyl positions and 13C-detected 13C-15N REDOR measurements were performed on peptide/antibody complexes of these labeled samples. Such hydrogen bonding was previously observed in a crystal structure of the V3 loop peptide/antibody complex RP142/59.1 [Ghiara et al. (1994) Science, 264, 82-85], but is shown by the REDOR measurements to be absent in the RP135/0.5beta complex. These results confirm the antibody-dependent conformational differences in the GPGR motif suggested by previously reported solid state NMR measurements of phi and psi backbone dihedral angles in the RP135/0.53 complex. In addition, we describe REDOR measurements on the helical synthetic peptide MB(i+4)EK in frozen solution that establish our ability to detect 13C-15N dipole-dipole couplings in the distance range appropriate to these hydrogen bonding studies. We also report the results of molecular modeling calculations on the central portion RP135, using a combination of the solid state NMR restraints of Weliky et al. [Nat. Struct. Biol., 6, 141-145, 1999] and the liquid state NMR restraints of Tugarinov et al. (Nat. Struct. Biol., 6, 331-335, 1999]. The dynamics calculations demonstrate the mutual compatibility of the two sets of experimental structural restraints and reduce ambiguities in the solid state NMR restraints that result from symmetry and signal-to-noise considerations. PMID- 10826884 TI - Characterization of surfactant liquid crystal phases suitable for molecular alignment and measurement of dipolar couplings. AB - Media employed for imparting partial alignment onto solute molecules have recently attracted considerable attention, since they permit the measurement of NMR parameters for solute biomolecules commonly associated with solid state NMR. Here we characterize a medium which is based on a quasi-ternary surfactant system comprising cetylpyridinium bromide/hexanol/sodium bromide. We demonstrate that dilute solutions of this system can exist in liquid crystalline phases which orient in the magnetic field and allow the measurement of residual dipolar couplings under a variety of conditions. The present system is extremely versatile and robust, tolerating different buffer conditions, temperature ranges and concentrations. PMID- 10826885 TI - A simple and inexpensive preparation of perdeuterated sorbitol for use as a biomacromolecule stabilization agent in NMR studies. AB - Sorbitol is an excellent protein-stabilization agent, but it is typically used at high concentrations where the 1H signals can interfere with NMR data collection and analysis. Deuteration of sorbitol can ameliorate this problem; however, perdeuterated sorbitol is not commercially available. We describe a simple and inexpensive method for preparation of perdeuterated sorbitol from perdeuterated glucose. The method is described explicitly and examples are given where the use of perdeuterated sorbitol has allowed the extraction of information, from NMR spectra, that is otherwise unobtainable. PMID- 10826886 TI - NMR experiments for the sign determination of homonuclear scalar and residual dipolar couplings. AB - A modified version of the JHH-TOCSY experiment, 'signed COSY', is presented that allows the determination of the sign of residual dipolar 1H-1H coupling constants with respect to the sign of one-bond 1H-X coupling constants in linear three-spin systems X-1H-1H, where X = 13C or 15N. In contrast to the original JHH-TOCSY experiments, the signs of J(HH) couplings may be determined for CH2-CH2 moieties and for uniformly 13C/15N-labelled samples. In addition, sensitivity is enhanced, diagonal peaks are suppressed and cross peaks are observed only between directly coupled protons, as in a COSY spectrum. PMID- 10826887 TI - Sequence-specific 1H, 13C and 15N resonance assignments and secondary structure of [2Fe-2S] ferredoxin from Halobacterium salinarum. PMID- 10826888 TI - Backbone H(N), N, Calpha, C' and Cbeta assignments of the 19 kDa DHFR/NADPH complex at 9 degrees C and pH 7.6. PMID- 10826889 TI - Letter to the editor: 1H, 15N and 13C resonance assignments for the catalytic domain of the yeast E2, UBC1. PMID- 10826890 TI - Complete 1H, 15N and 13C assignment of the functional domain of paracoccus denitrificans cytochrome c552 in the reduced state. PMID- 10826891 TI - Sequential NMR assignment of the RAS-binding domain of Byr2. PMID- 10826892 TI - Backbone resonance assignment of the N-terminal 24 kDa fragment of the gyrase B subunit from S. aureus complexed with novobiocin. PMID- 10826893 TI - Assignment of 1H, 13C and 15N NMR signals in the toluene 4-monooxygenase effector protein. PMID- 10826894 TI - Hyperlipidemia associated with HIV protease inhibitor use: pathophysiology, prevalence, risk factors and treatment. AB - Despite potent antiretroviral activity, the HIV-1 protease inhibitors have recently been associated with abnormal serum lipoprotein concentrations. The purpose of this review is to describe serum lipid abnormalities related to protease inhibitor use. A MEDLINE search up to June 1, 1999, and abstracts from recent scientific meetings were primary data sources. Lipid disturbances in HIV infected patients receiving protease inhibitors generally consist of elevated triglycerides and total cholesterol levels; HDL cholesterol is often reduced. The pathophysiological mechanism by which the protease inhibitors induce these lipid abnormalities has been hypothesized, but is unknown. Cases of pancreatitis and coronary heart disease have been described in hyperlipidemic patients receiving protease inhibitors. Treatment of protease inhibitor-related hyperlipidemia is unknown. Exchanging the offending protease inhibitor for nevirapine may be helpful in certain patients. Atorvastatin in combination with gemfibrozil has been used with limited success in a small number of individuals. PMID- 10826895 TI - Clinical aspects of nephropathia epidemica (Puumala virus infection) in Europe: a review. AB - Nephropathia epidemica (NE) is a prevalent zoonosis throughout Europe and is caused by the Puumala type of hantavirus. The incidence of NE varies in a cyclic fashion, with peaks occurring every 3rd to 4th year, coinciding with peaks in vole populations. The clinical course of NE is generally milder than haemorrhagic fever with renal syndrome caused by hantaviruses in other parts of the world. Typically, NE has a sudden onset with fever, headache, backpain and gastrointestinal symptoms. However, severe complications, e.g. gastrointestinal haemorrhage, occur and fatal cases have been reported. Renal involvement is prominent and manifests as initial oliguria and later as marked polyuria. Tests of renal function show pronounced glomerular and tubular involvement. Vaccine against Puumala virus infection as well as specific treatment for NE are still lacking. PMID- 10826896 TI - Clinical syndromes associated with adult pneumococcal cellulitis. AB - Streptococcus pneumoniae is an uncommonly recognized etiology of cellulitis in adults. A review of the literature uncovered 30 cases of pneumococcal skin infection in adults. Typically, all patients with pneumococcal cellulitis had an underlying chronic illness, or were immunocompromised by drug or alcohol abuse. Pneumococcal cellulitis presents as two distinctive clinical syndromes: one with extremity involvement in individuals with diabetes and substance abuse; and a second involving the head, neck and upper torso in individuals with systemic lupus erythematosis, nephrotic syndrome and hematologic disorders. For each there are statistically significant associations between the location of pneumococcal cellulitis and underlying clinical disorders. In contrast to other common bacterial etiologies, pneumococcal cellulitis is frequently associated with blood stream invasion, tissue necrosis and suppurative complications. Patients often require surgical interventions and prolonged hospitalizations. A high degree of suspicion and early aggressive management is needed for those presenting with cellulitis characterized by bullae and violaceous color. PMID- 10826897 TI - Screening and diagnosis of congenital cytomegalovirus infection: a 5-y study. AB - Although cytomegalovirus infection is the most common infection transmitted via the placenta, there are no guidelines for routine screening to detect children congenitally infected with cytomegalovirus. From 1993 to 1997, maternal serum and cord vein blood of newborns was screened for HCMV-IgM (n = 21,183). Urine was examined for HCMV-excretion during the first postnatal week to prove HCMV infection in children who expressed HCMV-IgM in cord vein blood (n = 13) or who were born to mothers positive for HCMV-IgM in the serum (n = 234), or when both cord vein blood and maternal serum were positive for HCMV-IgM (n = 6). Congenital HCMV infection was detected in 17 newborns. To determine the incidence of congenital HCMV infection, only those mother/child pairs were selected in whom serum and cord vein blood were investigated (n = 5967 mother/child pairs). In this group 13 newborns were infected. The observed incidence for congenital HCMV infection is 0.21%. It is concluded that that this screening programme will detect those children at risk for congenital HCMV infection. These children have to be examined for virus excretion in the urine. Although the observed incidence is only 0.21%, congenital HCMV infection is a problem that can no longer be neglected because of its long-term sequelae. PMID- 10826898 TI - Hepatitis C virus serotypes in haemodialysis patients in South-East Italy. AB - Hepatitis C virus (HCV) infection is highly prevalent in haemodialysis patients. To date, only a few studies involving a small number of subjects have characterized HCV-infected dialysis patients by serotyping. The spread of HCV serotypes in 114 HCV-positive dialysis patients from the same geographical area was evaluated by Murex HCV serotyping assay. Serotypes were detected in 102 subjects (89.5%), with type 1 being the most frequent (37.7%), followed by types 2 (19.3%), 4 (8.8%) and 3 (7.9%). Types 5 and 6 were the least prevalent (3.5%). Ten samples (8.8%) revealed mixed infections: type 1 was detectable in all and the co-infecting HCV types were types 2, 3 and 4 in 3, 4 and 3 cases, respectively. These results suggest that the serotyping assay as an alternative method of distinguishing the major types of HCV, also for particular risk groups and especially in laboratories that lack the specific expertise to perform genotyping methods. Age-related differences in patients with type 5 compared with those with types 3 and 6 may provide evidence of a more recent spread of these latter types. PMID- 10826899 TI - Hepatocellular carcinoma in Sweden: its association with viral hepatitis, especially with hepatitis C viral genotypes. AB - Viral markers of chronic hepatitis were tested for in 95 frozen serum samples from 299 patients from Malmo, Sweden, with hepatocellular carcinoma (HCC), diagnosed between 1977 and 1994. Hepatitis B analysis included anti-HBc, HBsAg and, if anti-HBc positive, HBV DNA. Hepatitis C infection analysis included anti HCV screening, RIBA, HCV RNA and HCV genotyping. HCV genotyping was also carried out in 9 HCV-viraemic HCC-patients from Gothenburg. HCV genotype distribution in HCC cases was compared with Swedish HCV-infected blood donors. Among the 95 patients from Malmo, 28 (29%) had anti-HBc, but only 5 (5%) were chronic HBV carriers, compared with 16 (17%) with chronic hepatitis C (p = 0.021). HCV related HCC was more common among immigrants (8/16 vs. 8/79; p < 0.001). Genotyping of 25 HCV-infected cases showed genotype 1a in 6 (24%), genotype 1b in 13 (52%), genotype 2b in 4 (16%), and genotype 3a in 2 (8.0%) patients. Genotype 1b was more common among HCC patients than among blood donors (p < 0.001), but 8 of 13 genotype 1b-infected patients were from countries where genotype 1b is predominant. Among native Swedes there was no difference between the HCV genotypes infecting blood donors and those found in HCC patients. PMID- 10826900 TI - Subcutaneous interleukin-2 in combination with anti-retroviral therapy for treatment of HIV-1-infected subjects. AB - A total of 11 HIV-1 positive patients, with CD4+ cell counts between 200 and 500/microl, who were in stable anti-retroviral therapy, were treated with subcutaneous recombinant human IL-2 thrice weekly administered on an out-patient basis in a dose-escalating manner. Subcutaneous IL-2 was well tolerated and associated with only mild to moderate constitutional symptoms and local inflammation at the injection site. CD4+ cell count increased from 404 +/- 48/microl at baseline to 639 +/- 88/microl at week 6, with proportionate increases in naive cells and memory cells. Increased doses of IL-2 were then needed to sustain the number of CD4+ cells. After discontinuation of IL-2 treatment, CD4+ cell count returned to baseline levels. IL-2 induced a reduction in the percentage of CD8+ CD38+ and CD8+ HLA-DR+ cells, an increase in the fraction of CD8+ CD25+ and CD8+ CD122+, and an elevation in the number of NK cells. IL-2 did not induce any clinically significant change in plasma HIV-RNA. In conclusion, IL-2 can safely be administered subcutaneously on an out-patient basis to HIV-infected individuals with CD4+ cell counts from 200/microl to 500/microl and with some improvement in immunological abnormalities. Continuous therapy, however, seems to result in the development of tachyphylaxia. PMID- 10826901 TI - Decreased GATA3 mRNA expression in human T-cell lymphotropic virus type 1 (HTLV 1) infection. AB - GATA3 is a specific T-cell transcription factor involved in the expression of T cell receptor (TCR). In order to characterize the relationship between HTLV-1 infection, which has been reported to be associated with down-regulation of genes belonging to the TCR/CD3 complex, and the transcription factor GATA3, we evaluated, by semi-quantitative RT-PCR, the expression of GATA3 gene in HTLV-1 carriers and individuals with related diseases. The study included 4 asymptomatic carriers, 2 patients with adult T-cell leukaemia/lymphoma (ATLL), 1 patient with HTLV-1 associated myelopathy (HAM)/tropical spastic paraparesis (TSP) and 7 healthy blood donors. A considerable decrease in the expression of the GATA3 mRNA was observed in all subjects infected by HTLV-1 and no expression of GATA3 mRNA was observed in 1 subject with ATLL and in 1 with HAM/TSP. PMID- 10826902 TI - Vertical transmission of HIV-1 in Poland. AB - The aim of our study was to evaluate changes in vertical transmission of HIV infection in Poland after introducing zidovudine prophylactic strategies. Data from the Department of Children's Infectious Diseases (a paediatric HIV referral centre) at the Medical University, Warsaw was studied. Since 1989 vertical transmission of HIV-1 has been studied in 100 children born to 91 HIV-positive mothers (2 sets of twins). Zidovudine therapy, mode and timing of delivery and their relationship to perinatal HIV-1 infection were analysed. From 1989 to 1994 the transmission rate was 31.5%. Since 1995, when recommendations based on ACTG 076 were issued, a decline in a transmission rate to 19.6% was reported. 62% (32 out of 52) mother-infant pairs received zidovudine therapy. None of those children have become HIV infected. Zidovudine chemoprophylaxis regimen reduces the risk for mother to child transmission. It should be recommended for all HIV infected pregnant women or women in labour and their infants. PMID- 10826903 TI - Risk factors for HIV-2 seropositivity among older people in Guinea-Bissau. A search for the early history of HIV-2 infection. AB - Because the seroprevalence of HIV-2 has been shown to be high in older age groups, we conducted a survey of all persons aged 50 years or over in two districts in Bissau, investigating the presence of HIV antibodies and possible risk factors for HIV infection with a particular emphasis on age, the impact of the war of independence (1963-74), traditional marital and extramarital sex patterns, blood contact and contact with monkeys. In 670 participants, the HIV-2 prevalence was 14.3%; 16.1% in women and 12.3% in men. The HIV-1 prevalence was only 0.5% (3/670). The HIV-2 prevalence peaked for men in the 60-69 years age group, and for women in the 50-59 years age group, declining markedly in the following age group for both men and women (OR = 0.09 (0.01-0.51), OR = 0.37 (0.15-0.82), respectively). This pattern could be due to differential mortality for HIV-2 infected individuals or to a cohort effect for a generation who were sexually active at the time of the war of independence in the 1960s and early 1970s in Bissau. Supporting the link with the colonial army, women who had had sex with a white man had a higher seroprevalence (OR = 3.63 (1.12-11.24)). The ethnic group indigenous to Bissau city had a much lower prevalence, but demographic and cultural risk factors such as marital status, religion, education and having lived outside Bissau were not associated with HIV-2. In the multivariate analyses for women, variables related to extramarital sex or prostitution (having sex with a white man, having lived in Senegal, not living with husband, and not marrying first sexual partner) were associated with higher risk. For men, previous spouses who had died or had divorced were associated with higher prevalence. Having married the first sexual partner was protective against HIV-2 infection for both men (OR = 0.29 (0.09-0.76)) and women (OR = 0.19 (0.04 1.00)). Hospitalizations, possibly due to transfusions, tended to be associated with higher risk, but only for women (OR = 1.83 (0.97-3.48)). The focus of the HIV-2 epidemic in Guinea-Bissau is likely to be endogenous, and the war of independence and the colonial army with its associated prostitution may have played an important part in propagating the virus. PMID- 10826904 TI - Seroprevalence of antibodies to Chlamydia pneumoniae in elderly people: a two decade longitudinal and cohort difference study. AB - A gerontological population of 178 men and 249 women was investigated regarding the prevalence of antibodies specific to Chlamydia pneumoniae. A longitudinal substudy was carried out on 22 men and 44 women, age range 70-90 y. Antibodies specific to C. pneumoniae were common. Men had higher prevalences and higher antibody levels than women. More than half of the individuals in the longitudinal study had significant IgG and/or IgA titre changes (> or = 4-fold) between the ages of 70 and 90 y, suggesting that C. pneumoniae infections are common in the elderly population. This is of importance for the treatment of respiratory infections in elderly people. PMID- 10826905 TI - Hospitalization due to Pasteurella multocida-infected animal bite wounds: correlation with inadequate primary antibiotic medication. AB - Over a 10-y period, patients hospitalized with Pasteurella-induced cat or dog bite-associated wound infection were analysed retrospectively with regard to preceding antibiotic medication. In 10/14 cases, hospitalization was necessitated in spite of prophylactic or therapeutic administration of oral antibiotics. In 1 case, phenoxymethylpenicillin and flucloxacillin had been prescribed. The other patients received flucloxacillin (7 patients), erythromycin, or cefadroxil (1 patient each), agents that are not consistently active against Pasteurella. In conclusion, hospitalization due to Pasteurella-induced animal bite-associated wound infection seemed to be related to the prescription of suboptimal oral antibiotic therapy at a preceding medical consultation. PMID- 10826906 TI - Molecular characteristics and susceptibility to antibiotics of serogroup A Neisseria meningitidis strains isolated in Senegal in 1999. AB - A total of 22 strains of Neisseria meningitidis isolated from cerebrospinal fluid samples of patients in Dakar, Senegal, in the course of an epidemic of meningococcal meningitis in 1999 were studied. All the strains were serogroup A, serotype 21:P1.9 and belonged to clonal subgroup III-1. The strains were resistant to sulphonamide, but were susceptible to ampicillin, ceftriaxone and chloramphenicol, which are used in the treatment of cerebrospinal meningitis in Senegal. PMID- 10826907 TI - Influence of breastmilk on the development of resistance to intestinal colonization in infants born at the Atma Jaya Hospital, Jakarta. AB - A study of intestinal colonization resistance (CR) in breastfed versus formula fed newborns at 4 intervals after birth in Jakarta, Indonesia, is described. To measure the intestinal CR for gram-negative enterobacilli, mean values of Enterobacteriaceae concentrations and mean numbers of Enterobacteriaceae biotypes were determined. The CR values found in this study show, that in all 4 sampling periods, at < 1, 2, 4 and 6 months, the mean concentration of Enterobacteriaceae was somewhat lower in the breastfed group than in the formula-fed group (only significant at 6 months). This means that the intestinal CR of the breastfed group may have been slightly higher than that in the formula-fed group. In both study groups, the CR was lower in the second and fourth month than soon after birth and at 6 months. For epidemiological reasons, comparison was performed of the Enterobacteriaceae biotypes found in samples from mother and child. The data show that, in the first sampling period, regardless of the theoretical possibility of a 'more intense (skin) contact' during breastfeeding (which might promote transfer of also microorganisms), the breastfed infants had a significantly lower percentage of identical Enterobacteriaceae biotypes than did the formula-fed group. This could possibly be ascribed to a higher CR in the breastfed group. Determination of the concentration of Enterococcus species was found applicable to reproducibly measure the CR in the newborns at 6 months and in the mother-group. PMID- 10826908 TI - Ceftazidime versus aztreonam in the treatment of pseudomonal chronic suppurative otitis media in children. AB - In a prospective, open, randomized trial, ceftazidime was given to 15 children and aztreonam to another 15 children with chronic suppurative otitis media without cholesteatoma (CSOM). Patients enrolled to the study had a pure culture of Pseudomonas aeruginosa growing in middle ear discharge. The subjects' mean age was 56 months in the ceftazidime group and 48 months in the aztreonam group. Success rate, defined as complete disappearance of discharge, was 84.6% in the ceftazidime treated patients and 67% in the aztreonam group (p value not significant). The number of days required for complete dryness of the ear was 7.9 and 8.4 respectively. Two patients in each group had recurrence of suppurative discharge within 90 d from discontinuation of antibiotic treatment. These results suggest that aztreonam is an optional alternative systemic treatment for paediatric patients with pseudomonal CSOM, especially in subjects allergic to penicillin. PMID- 10826909 TI - Risk factors predicting complications in blood culture-proven typhoid fever in adults. AB - To create a prognostic model for complications of blood culture-proven typhoid fever in adults (> or = 15 y old), a retrospective cohort was assembled though review of the medical records of the hospitalized patients treated for typhoid fever over a 3-y period ending January 1995. Of the 59 patients included, 21 (35.6%) developed various complications of typhoid fever. No patient included died. Four baseline variables (abdominal pain, systolic blood pressure < 100 mmHg, hypoalbuminaemia < 32 g/l and laboratory evidence of disseminated intravascular coagulation) were independently associated with complications and were used to create a prognostic model. The prediction accuracy of the model was determined using the concordance index (c-index). The results (c-index, 0.915 [95%, CI, 89.0-93.0]) showed that the model predicted complications significantly better than chance. The model stratified patients into 3 prognostic stages: low risk for complications (0%; stage I), intermediate risk (42.9%; stage II) and high risk (92.3%; stage III) (p = 0.001). If validated in other settings, it will help clinicians in predicting complications in patients with blood culture-proven typhoid fever on admission. PMID- 10826910 TI - Shigellosis of childhood in northern Greece: epidemiological, clinical and laboratory data of hospitalized patients during the period 1971-96. AB - The aim of this study was to evaluate epidemiological, clinical and laboratory data of shigellosis in children from northern Greece, hospitalized in our department during the period 1971-96. In total, 422 cases of shigellosis, aged 1 month to 14 y (238M, 184F) were hospitalized during the study period. The annual distribution was approximately stable until 1990, the mean number of cases per year being about 20. During the last 4 y the incidence significantly decreased. Shigella was serotyped in 138/422 cases. Seventy six of the strains were S. flexneri (55%) and 56 S. sonnei (40%). In the majority of cases the clinical picture was mild. Severe dehydration was seen in only 6 patients. Ninety four patients (22%) had extra-intestinal manifestations. Most common of these were convulsions (16%) and, less frequently, disturbances of consciousness (n = 26), rash (n = 9), shock and disseminated intravascular coagulopathy (n = 2), nerve paralysis (n = 2), severe anaemia (n = 2) and haemolytic-uraemic syndrome (n = 1). Nine patients had acute encephalopathy of 12 h to 12 d duration. It is important to note that all these cases recovered completely with no residual neurological deficit, except for 1 girl who developed temporal epilepsy 8 y later. Spinal fluid was normal in all 42 examined patients. Antibiotics were given to 212 of 422 patients, mainly during the first half of the study period. Shigella resistance to antibiotic was significant for cotrimoxazole (24%) and ampicillin (16%). All patients were cured. Shigellosis is a mild disease in our area, with a decreasing prevalence. PMID- 10826911 TI - Polymyositis associated with primary cytomegalovirus infection. AB - A case of polymyositis associated with primary cytomegalovirus infection in a 17 y-old girl is reported. The girl exhibited fever, sore throat, progressive myalgia and muscle weakness with elevated creatine kinase, atypical lymphocytosis and myopathic features in the electromyogram. Histopathologically, biopsied muscle met the criteria for polymyositis. Primary cytomegalovirus infection was proven by seroconversion of IgG as well as IgM antibodies. This is the first report of an association between cytomegalovirus infection and polymyositis. PMID- 10826912 TI - Streptococcus mitis septicaemia and hepatitis. AB - The case of an otherwise well 9-y-old boy with fever, papular rash, jaundice and impaired liver function is presented. Streptococcus mitis sensitive to penicillin grew in blood culture. The boy had an excellent outcome. The clinical spectrum of viridans streptococci may be wider than currently anticipated, and Streptococcus mitis may cause septicaemia and hepatitis in immunocompetent individuals. PMID- 10826913 TI - Group B streptococcus meningitis in an HIV-positive adult: case report and review. AB - We report a new case of meningitis due to group B streptococci (S. agalactiae) in an HIV-positive patient. Only three other cases have been reported so far, and common features are discussed. PMID- 10826914 TI - Moraxella catarrhalis endocarditis: case report and review of the literature. AB - A case of bacterial endocarditis caused by Moraxella catarrhalis in an apparently immunocompetent Greek male is presented, which was diagnosed after a 2-month history of low-grade fever of unknown origin. The agent seems to be a rare pathogen, but due to the high mortality rate, it should always be considered in the differential diagnosis of relevant cases. Beta-lactamase production by many strains complicates the choice of antibiotic. PMID- 10826915 TI - A patient with suspected sarcoidosis died from miliary tuberculosis. AB - In Western Europe, awareness of tuberculous lymphadenitis (TLA) has declined. This report describes a patient who had suspected sarcoidosis for 3.5 y, who eventually died from miliary tuberculosis due to delay in diagnosis. The report includes a discussion of the differential diagnosis of TLA and sarcoidosis. PMID- 10826916 TI - Direct NAD(P)H hydrolysis into ADP-ribose(P) and nicotinamide induced by reactive oxygen species: a new mechanism of oxygen radical toxicity. AB - The effect of different oxygen radical-generating systems on NAD(P)H was determined by incubating the reduced forms of the pyridine coenzymes with either Fe2+-H2O2 or Fe3+-ascorbate and by analyzing the reaction mixtures using a HPLC separation of adenine nucleotide derivatives. The effect of the azo-initiator 2,2'-azobis(2-methylpropionamidine)dihydrochloride was also tested. Results showed that, whilst all the three free radical-producing systems induced, with different extent, the oxidation of NAD(P)H to NAD(P)+, only Fe2+-H2O2 also caused the formation of equimolar amounts of ADP-ribose(P) and nicotinamide. Dose dependent experiments, with increasing Fe2+ iron (concentration range 3-180 microM) or H2O2 (concentration range 50-1000 microM), were carried out at pH 6.5 in 50 mM ammonium acetate. NAD(P)+, ADP-ribose(P) and nicotinamide formation increased by increasing the amount of hydroxyl radicals produced in the medium. Under such incubation conditions NAD(P)+/ADP-ribose(P) ratio was about 4 at any Fe2+ or H2O2 concentration. By varying pH to 2.0, 3.0, 4.0, 4.5, 5.0, 5.5, 6.0, 7.0 and 7.4, NAD(P)+/ADP-ribose(P) ratio changed to 5.5, 3.2, 1.8, 1.6, 2.0, 2.5, 3.0, 5.4 and 6.5, respectively. Kinetic experiments indicated that 90-95% of all compounds were generated within 5s from the beginning of the Fenton reaction. Inhibition of ADP-ribose(P), nicotinamide and NAD(P)+ production of Fe2+-H2O2 treated NAD(P)H samples, was achieved by adding mannitol (10-50 mM) to the reaction mixture. Differently, selective and total inhibition of ADP-ribose(P) and nicotinamide formation was obtained by performing the Fenton reaction in an almost completely anhydrous medium, i.e. in HPLC-grade methanol. Experiments carried out in isolated postischemic rat hearts perfused with 50 mM mannitol, showed that, with respect to values of control hearts, this hydroxyl radical scavenger prevented reperfusion-associated pyridine coenzyme depletion and ADP ribose formation. On the basis of these results, a possible mechanism of action of ADP-ribose(P) and nicotinamide generation through the interaction between NAD(P)H and hydroxyl radical (which does not involve the C-center where "conventional" oxidation occurs) is presented. The implication of this phenomenon in the pyridine coenzyme depletion observed in postischemic tissues is also discussed. PMID- 10826917 TI - Trypanosoma cruzi dihydrolipoamide dehydrogenase is inactivated by myeloperoxidase-generated "reactive species". AB - Dihydrolipoamide dehydrogenase (LADH) from Trypanosoma cruzi was inactivated by treatment with myeloperoxidase (MPO)-dependent systems. With MPO/H2O2/NaCl, LADH lipoamide reductase and diaphorase activities significantly decreased as a function of incubation time. Iodide, bromide, thiocyanide and chloride effectively supplemented the MPO/H2O2 system, KI and NaCl being the most and the least effective supplements, respectively. LADH inactivation by MPO/H2O2/NaCl and by NaOCl was similarly prevented by thiol compounds such as GSH, L-cysteine, N acetylcysteine, penicillamine and N-(2-mercaptopropionyl-glycine) in agreement with the role of HOCI in LADH inactivation by MPO/H2O2/NaCl. LADH was also inactivated by MPO/NADH/halide, MPO/H2O2/NaNO2 and MPO/NADH/NaNO2 systems. Catalase prevented the action of the NADH-dependent systems, thus supporting H2O2 production by NADH-supplemented LADH. MPO inhibitors (4-aminobenzoic acid hydrazide, and isoniazid), GSH, L-cysteine, L-methionine and L-tryptophan prevented LADH inactivation by MPO/H2O2/NaNO2. Other MPO systems inactivating LADH were (a) MPO/H2O2/chlorpromazine; (b) MPO/H2O2/monophenolic systems, including L-tyrosine, serotonin and acetaminophen and (c) MPO/H2O2/di- and polyphenolic systems, including norepinephrine, catechol, nordihydroguaiaretic acid, caffeic acid, quercetin and catechin. Comparison of the above effects and those previously reported with pig myocardial LADH indicates that both enzymes were similarly affected by the MPO-dependent systems, allowance being made for T. cruzi LADH diaphorase inactivation and the greater sensitivity of its LADH lipoamide reductase activity towards the MPO/H2O2/NaCl system and NaOCl. PMID- 10826918 TI - Fructose induced deactivation of antioxidant enzymes: preventive effect of pyruvate. AB - Glycation initiated changes in tissue proteins, which are triggered by the Schiff base formation between the sugar carbonyl and the protein -NH2, have been suggested to play an important role in the development of diabetes-related pathological changes such as the formation of cataracts. While the initial reaction takes place by the interaction of >C=O of the parent sugars with the NH2 of proteins, reactive oxygen species (ROS) dependent generation of more reactive dicarbonyl derivatives from the oxidation of sugars also plays a significant role in these changes, altering the structural as well as functional properties of proteins. The purpose of this study was to examine whether the activities of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), catalase and superoxide dismutase (SOD) could be affected by the high levels of fructose prevalent in diabetic lenses. Incubation of the enzymes with this sugar led to a significant loss of their activities. GAPDH was inactivated within a day. This was followed by the inactivation of catalase (3-4 days) and SOD (6 days). The loss of the activities was prevented significantly by incorporation of pyruvate in the incubation mixture. The protective effect is ascribable to its ability to competitively inhibit glycation as well as to its ROS scavenging activity. Hence, it could play a significant role in the maintenance of lens physiology and cataract prevention. PMID- 10826919 TI - Pretreatment with methylprednisolone protects the isolated rat heart against ischaemic and oxidative damage. AB - Methylprednisolone (MP), a synthetic glucocorticoid, is widely used clinically and experimentally as acute antiinflammatory treatment. The molecular actions of MP indicate that pretreatment with this drug may be cardioprotective. We investigated if giving rats MP prior to excising their hearts for Langendorff perfusion protected cardiac function against oxidative stress, and if this was mediated by increasing antioxidant defence or influencing myocardial nitric oxide synthase (NOS). Rats (n=6-11 in each group) were injected with MP (40mg/kg i.m.) or vehicle 24 and 12 h before Langendorff-perfusion with 30 min global ischaemia and 60 min reperfusion, or 10 min perfusion with 180 micromol/L hydrogen peroxide. Other hearts were exposed to 30 min global ischaemia 5 days after MP injection. Additional hearts were sampled before, during, and after ischaemia for analyzing tissue activity of antioxidant enzymes. Tissue endothelial and inducible NOS (eNOS and iNOS) were investigated by immunoblotting and semiquantitative RT-PCR in a time-course after MP injection. Pretreatment with MP improved left ventricular function and increased coronary flow during postischaemic reperfusion, and this effect was sustained 5 days afterwards. When exposing hearts to hydrogen peroxide, MP improved coronary flow. Catalase, glutathione peroxidase, and oxidized glutathione were increased during reperfusion of MP-treated hearts compared to vehicle only. MP did not influence eNOS at protein or mRNA level. iNOS could not be detected by immunoblotting, indicating low cardiac enzyme content. Its mRNA initially increased the first hour after injection, thereafter decreased. In conclusions, pretreating rats with MP protects the heart against ischaemia-reperfusion dysfunction. This effect could be due to increase of tissue antioxidant activity during reperfusion. MP did not influence cardiac eNOS. mRNA for iNOS was influenced by MP, but the corresponding protein could not be detected. PMID- 10826920 TI - Pyruvate secreted by human lymphoid cell lines protects cells from hydrogen peroxide mediated cell death. AB - Reactive oxygen species (ROS) released from polymorphonuclear leukocytes and macrophages could cause DNA damage, but also induce cell death. Therefore inhibition of cell death must be an important issue for accumulation of genetic changes in lymphoid cells in inflammatory foci. Scavengers in the post culture medium of four lymphoid cell lines, lymphoblastoid cell lines (LCL), Raji, BJAB and Jurkat cells, were examined. Over 80% of cultured cells showed cell death 24 h after xanthine (X)/xanthine oxidase (XOD) treatment, which was suppressed by addition of post culture medium from four cell lines in a dose-dependent manner. H2O2 but not O2*- produced by the X/XOD reaction was responsible for the cytotoxity, thus we used H2O2 as ROS stress thereafter. The H2O2-scavenging activity of post culture media from four cell lines increased rapidly at the first day and continued to increase in the following 2-3 days for LCL, Raji and BJAB cells. The scavenging substance was shown to be pyruvate, with various concentrations in the cultured medium among cell lines. Over 99% of total pyruvate was present in the extracellular media and less than 1% in cells. alpha Cyano-4-hydroxycinnamate, a specific inhibitor of the H+-monocarbohydrate transporter, increased the H2O2-scavenging activity in the media from all four cell lines via inhibition of pyruvate re-uptake by cultured cells from the media. These findings suggest that lymphoid cells in inflammatory foci could survive even under ROS by producing pyruvate, so that accumulation of lymphoid cells with DNA damage is possible. PMID- 10826921 TI - Effect of age on the profile of alkanes in normal human breath. AB - Ethane and pentane in breath are markers of oxidative stress, produced by ROS mediated lipid peroxidation of n-3 and n-6 polyunsaturated fatty acids (PUFAs), but little is known about other n-alkanes in normal human breath. We investigated the spectrum of alkanes in normal human alveolar breath, and their variation with age. Fifty normal humans were studied (age range 23-75, median 35). Volatile organic compounds (VOCs) in alveolar breath were captured on sorbent traps and assayed by gas chromatography and mass spectroscopy. Alveolar gradients (concentration in breath minus concentration in ambient room air) of alkanes were determined. C4-C20 alkanes were observed in breath and room air. Their mean alveolar gradients were negative from C4 to C12 and positive from C13 to C20. The mean alveolar gradients of four alkanes (C5-C8) were significantly less negative in the older subjects (p < 0.05). There were no significant differences between males and females. Normal human breath contained a spectrum of alkanes which may include new markers of oxidative stress. The mean rate of clearance (via cytochrome p450) exceeded the mean rate of synthesis (by ROS-mediated oxidative stress) for C4-C12 alkanes, while synthesis was greater than clearance for C13 C20 alkanes. The elevated alkane profile in older subjects was consistent with an age-related increase in oxidative stress, though an age-related decline in alkane clearance rate may have contributed. PMID- 10826922 TI - Gain in functions of mutant Cu,Zn-superoxide dismutases as a causative factor in familial amyotrophic lateral sclerosis: less reactive oxidant formation but high spontaneous aggregation and precipitation. AB - Eight mutant Cu,Zn-superoxide dismutases (SODs) related to familial amyotrophic lateral sclerosis (FALS) were produced in a baculovirus/insect cell expression system and their molecular properties in terms of hydroxyl radical formation and aggregation were compared with the wild-type enzyme. Treatment of the enzymes with Chelex 100 resin decreased Cu contents as well as SOD activities in all mutant Cu,Zn-SODs, indicating that the affinities of the enzymes for copper ion were decreased. Contrary to previous reports, all the mutant Cu,Zn-SODs exhibited less reactive oxidant producing ability in the presence of hydrogen peroxide than the wild-type enzyme. Both SOD activities and their reactive oxidant forming correlated well with the copper ion content of the molecules. In addition, the proteins spontaneously aggregated and were precipitated by simple centrifugation at 12,000g for 20 min in keeping their enzyme activities. Since hyaline inclusions found in FALS patients with SOD1 mutations contained components which were reactive to anti-Cu,Zn-SOD antibody, a primary reaction caused by mutant SOD1 may be attributed to their propensity to form aggregates. Aggregated but still active mutant SOD1 would be expected to mediate the formation of reactive oxygen species and nitrosylation in a more condensed state. PMID- 10826923 TI - "Spin-labelling" with the second hydrogen radioisotope: radicals formed from squalene by muonium addition. AB - By radio-labelling with muonium (the second hydrogen radioisotope), a free radical species has been clearly identified in squalene (2,6,10,15,19,23 hexamethyl-2,6,10,14,18,22-tetracosahexaene ) with a muon-electron hyperfine coupling of 240.7 MHz, as measured by the muon spin rotation (MuSR) technique. The radical is undetectable in pure squalene due to its high viscosity and the large molecular size which leads to extreme line broadening, but its signals may be resolved on reducing the viscosity of the medium by dilution with diethyl ether. The potential of the MuSR method is thus demonstrated as a means for spin labelling radical species such as are formed from cellular antioxidants; ESR spectroscopy is unsuitable for studies of radicals formed from this molecule, due to extreme line-broadening and spectral complexity. PMID- 10826924 TI - Ceruloplasmin enhances DNA damage induced by hydrogen peroxide in vitro. AB - Ceruloplasmin (Cp) was found to promote the oxidative damage to DNA, as evidenced by the formation of 8-hydroxy-2'-deoxyguanosine and strand breaks, when incubated with H2O2 in vitro. The capacity of Cp to enhance oxidative damage to DNA was inhibited by hydroxyl radical scavengers such as sodium azide and mannitol, a metal chelator, diethylenetriaminepentaacetic acid, and catalase. Although the oxidized protein resulted in an increase in the content of carbonyl groups, the ferroxidase activity and the proteolytic susceptibility were not significantly altered. The release of a portion of Cu from Cp was observed, and conformational alterations were indicated by the changes in fluorescence spectra. Based on these results, we suggest that damage to DNA is mediated in the H2O2/Cp system via the generation of *OH by released Cu2+ and/or loosely bound Cu exposed from oxidatively damaged Cp through the conformational change. The release of Cu from Cp during oxidative stress could enhance the formation of reactive oxygen species and could also potentiate cellular damage. PMID- 10826925 TI - Reduction in free-radical-induced DNA strand breaks and base damage through fast chemical repair by flavonoids. AB - This paper provides evidence that dietary flavonoids can repair a range of oxidative radical damages on DNA, and thus give protection against radical induced strand breaks and base alterations. We have irradiated dilute aqueous solutions of plasmid DNA in the absence and presence of flavonoids (F) in a "constant *OH radical scavenging environment", k of 1.5 x 10(7) s(-1) by decreasing the concentration of TRIS buffer in relation to the concentration of added flavonoids. We have shown that the flavonoids can reduce the incidence of single-strand breaks in double-stranded DNA as well as residual base damage (assayed as additional single-strand breaks upon post-irradiation incubation with endonucleases) with dose modification factors of up to 2.0+/-0.2 at [F] < 100 microM by a mechanism other than through direct scavenging of *OH radicals. Pulse radiolysis measurements support the mechanism of electron transfer or H* atom transfer from the flavonoids to free radical sites on DNA which result in the fast chemical repair of some of the oxidative damage on DNA resulting from *OH radical attack. These in vitro assays point to a possible additional role for antioxidants in reducing DNA damage. PMID- 10826926 TI - Resveratrol inhibition of lipid peroxidation. AB - To define the molecular mechanism(s) of resveratrol inhibition of lipid peroxidation we have utilized model systems that allow us to study the different reactions involved in this complex process. Resveratrol proved (a) to inhibit more efficiently than either Trolox or ascorbate the Fe2+ catalyzed lipid hydroperoxide-dependent peroxidation of sonicated phosphatidylcholine liposomes; (b) to be less effective than Trolox in inhibiting lipid peroxidation initiated by the water soluble AAPH peroxyl radicals; (c) when exogenously added to liposomes, to be more potent than alpha-tocopherol and Trolox, in the inhibition of peroxidation initiated by the lipid soluble AMVN peroxyl radicals; (d) when incorporated within liposomes, to be a less potent chain-breaking antioxidant than alpha-tocopherol; (e) to be a weaker antiradical than alpha-tocopherol in the reduction of the stable radical DPPH*. Resveratrol reduced Fe3+ but its reduction rate was much slower than that observed in the presence of either ascorbate or Trolox. However, at the concentration inhibiting iron catalyzed lipid peroxidation, resveratrol did not significantly reduce Fe3+, contrary to ascorbate. In their complex, our data indicate that resveratrol inhibits lipid peroxidation mainly by scavenging lipid peroxyl radicals within the membrane, like alpha-tocopherol. Although it is less effective, its capacity of spontaneously entering the lipid environment confers on it great antioxidant potential. PMID- 10826927 TI - Bimodal RR interval distribution in chronic atrial fibrillation: impact of dual atrioventricular nodal physiology on long-term rate control after catheter ablation of the posterior atrionodal input. AB - INTRODUCTION: Radiofrequency (RF) catheter modification of the AV node in patients with atrial fibrillation (AF) is limited by an unpredictable decrease of the ventricular rate and a high incidence of permanent AV block. A bimodal RR histogram has been suggested to serve as a predictor for successful outcome but the corresponding AV node properties have never been characterized. We hypothesized that a bimodal histogram indicates dual AV nodal physiology and predicts a better outcome after AV node modification in chronic AF. METHODS AND RESULTS: Thirty-seven patients were prospectively subdivided into two groups according to the RR histogram of 24-hour ECG monitoring. Before to RF ablation, internal cardioversion and programmed stimulation were performed. Among the 22 patients (group I) with a bimodal RR histogram, dual AV nodal physiology was found in 17 (77%) patients. Ablation significantly decreased ventricular rate with loss of the peak of short RR cycles after ablation (mean and maximal ventricular rates: 32% and 35% rate reduction, respectively; P < 0.01). In 15 patients with a unimodal RR histogram (group II), dual AV nodal physiology was found in 2 (13%), and rate reductions were 16% and 17%, respectively. At 6 months, 3 (14%) patients in group I and 6 (40%) in group II underwent elective AV nodal ablation with pacemaker implantation due to intolerable rapid ventricular response to AF. CONCLUSION: Bimodal RR interval distribution during chronic AF suggests the presence of dual AV nodal physiology and predicts a better outcome of RF ablation of the posterior atrionodal input. PMID- 10826928 TI - Relevance of atrioventricular nodal physiology in patients with medically refractory atrial fibrillation. PMID- 10826930 TI - Spatial and temporal heterogeneity of depolarization and repolarization may complicate implantable cardioverter defibrillator therapy in Brugada syndrome. AB - INTRODUCTION: Dynamic variations in electrophysiologic phenomena inherent to the Brugada syndrome may complicate therapy with implantable cardioverter defibrillators (ICDs). METHODS AND RESULTS: Between 1997 and 1999, 3 of 7 patients with Brugada syndrome (1 man and 2 women, mean age 42 years) received an ICD. During follow-up, 2 patients experienced multiple inappropriate shocks. Simultaneously with dynamic changes in the surface ECG, endocardial ECGs revealed a dynamic decrease in the right ventricular R wave and an increase in the corresponding T wave, resulting in T wave oversensing. With ajmaline administration, these dynamic changes in endocardial signals were reproducible at different right ventricular sites, whereas left ventricular epicardial signals remained stable. Incremental AAI pacing and exercise stress testing resulted in similar changes in right ventricular endocardial signals, but normalization of the surface ECG apart from progressively increasing S waves in leads II, V5, and V6. Orciprenaline administration had no effect on ECG phenomena. After implantation of a left ventricular epicardial lead for sensing and pacing, no inappropriate tachycardia detection recurred. CONCLUSION: These findings demonstrate that, in Brugada syndrome, spontaneous or ajmaline-induced changes in the surface ECG may be paralleled by significant variations in the right ventricular endocardial electrogram that may result in ICD malfunction. Implantation of a left ventricular epicardial lead for sensing and pacing may be the ultimate successful approach in certain patients. To assure proper ICD function, ajmaline testing during ICD implantation appears to be helpful. PMID- 10826929 TI - Dependence of electrogram duration in right posteroseptal atrium and atrium pulmonary vein junction on pacing site: mechanism and implications regarding atrioventricular nodal reentrant tachycardia and paroxysmal atrial fibrillation. AB - INTRODUCTION: The fractionated atrial electrogram, a signal helpful in identifying the target site for radiofrequency catheter ablation of the slow AV nodal pathway, is considered to arise from nonuniform anisotropic electrical activity. However, the effects of pacing sites and radiofrequency ablation on these electrograms are not clear. Similarly, the nature of the fractionated atrial electrogram in the atrium-pulmonary vein junction has yet to be determined. METHODS AND RESULTS: Two experiments were performed in this study. Experiment 1 evaluated the fractionated atrial electrogram at target sites before and after slow AV nodal pathway ablation during sinus rhythm or during pacing from different sites. Group 1A consisted of 16 patients with dual AV nodal pathway physiology and AV nodal reentrant tachycardia who underwent successful ablation without residual slow AV nodal pathway. Group 1B consisted of 7 patients who underwent successful elimination of AV nodal reentry but with residual dual AV nodal pathway physiology. Group 1C consisted of 6 patients who still had AV nodal reentrant tachycardia after two applications of radiofrequency energy. In group 1D, there were 16 patients with dual AV nodal pathway physiology, but without inducible AV nodal reentrant tachycardia. In group 1E, there were 15 patients without dual AV nodal pathway physiology. Experiment 2 investigated the fractionated atrial electrogram in the ostium of the left and right superior pulmonary veins in 18 patients with paroxysmal atrial fibrillation (2A) and in 8 patients without paroxysmal atrial fibrillation (2B). Before radiofrequency ablation, electrogram duration in the right posteroseptal atrium during pacing from the middle coronary sinus or the right posterolateral atrium was shorter than that during pacing from the high right atrium (HRA) in all group 1 patients. After the successful elimination of the slow AV nodal pathway conduction in group 1A, atrial electrogram duration during HRA pacing was shorter than that before ablation. In experiment 2 patients, electrogram duration during pacing from the proximal or distal coronary sinus was shorter than that during pacing from HRA or sinus rhythm. CONCLUSION: These findings suggest that the fractionated atrial electrograms in the right posteroseptal atrium and ostium of left or right superior pulmonary veins are potentially consistent with nonuniform anisotropic propagation. Alternations of electrogram characteristics after successful radiofrequency ablation of the slow AV nodal pathway may arise from the changes of nonuniform anisotropic activity in the right posteroseptal atrium. PMID- 10826931 TI - Role of the input/output relation of sinoatrial myocytes in cholinergic modulation of heart rate variability. AB - INTRODUCTION: Modulation of sinus rate may be viewed as the transduction of an input signal (receptor stimulation) into an output signal (cycle length [CL]) by the sinus node. This study analyzes the input/output (I/O) relation of sinoatrial pacemaking elements and tests its impact on cholinergic modulation of heart rate variability. METHODS AND RESULTS: Spontaneous activity of isolated rabbit sinoatrial myocytes was recorded by patch clamp techniques. CL and all the action potential parameters determining CL were automatically measured from >100 consecutive cycles. Acetylcholine (ACh, 5 to 50 nM) increased CL by decreasing diastolic depolarization rate (DDR) only. This was associated with a substantial increase in the coefficient of variation of CL and minor changes in the coefficient of variation of other parameters. A simple function relating CL to action potential parameters accurately described CL response to ACh (I/O relation). Numerical simulations based on this I/O relation showed that ACh induced depression of DDR might, by its own, increase CL variability. CONCLUSION: Time-domain measurements of CL variability may not necessarily reflect variability of the neural input to the sinoatrial node, but also may be affected by its tonic level. Properties of the I/O relation of sinoatrial myocytes may fully account for the dependency of CL variability on mean heart rate, previously described in humans. Any condition depressing DDR may enhance CL variability, independent of changes in the pattern of neural activity. PMID- 10826932 TI - Cycle length dynamics at the onset of postinfarction ventricular tachycardias induced in canines: dependence on interval-dependent excitation properties of the reentrant substrate. AB - INTRODUCTION: Postinfarction monomorphic ventricular tachycardias induced by programmed stimulation may display initial cycle length (CL) variations before stabilizing. METHODS AND RESULTS: To show that tachycardia onset dynamics depend on rate-dependent electrical properties of the reentrant substrate, we extracted activation times and maximum negative slopes of local activation complexes ( dV/dt(max)) from 191 unipolar electrograms recorded in the anterior left ventricular wall of anesthetized, 3-day-old infarct canine preparations. Measurements were made of the responses to programmed stimulation, as well as in early and later beats of tachycardias, which displayed either a constant trend in CL (group A, n = 5 preparations) or one in which CL prolongation occurred according to an exponential course before stabilizing (group B, n = 9). Stimulation protocols inducing the tachycardias were more aggressive and their CL was significantly shorter (CL = 159 +/- 24 msec) in group A than in group B (stabilized CL = 206 +/- 34 msec). Reentrant activity occurred in subepicardial areas in which the absolute value of -dV/dt(max) (absolute value(-dV/dtmax)) was heterogeneously depressed (<2 mV/msec). Absolute value(-dV/dtmax) was reduced and activation delay increased in the successive responses to extrastimuli. Further reductions in absolute value(-dV/dtmax) (10% to 23%) were shown to occur between early and later beats in 5 of the 9 tachycardias in group B (no change in the 4 others), and they were associated with localized prolongation of conduction times in reentrant pathways. In contrast, absolute value(-dV/dtmax) improved in all group A tachycardias (7% to 25%). CONCLUSION: This study provides evidence that the onset dynamics of postinfarction ventricular tachycardias are determined by interval-dependent electrical changes occurring in the reentrant substrate. PMID- 10826933 TI - Cycle length dynamics during ventricular tachycardia in the infarcted heart. PMID- 10826934 TI - Simultaneous optical mapping of transmembrane potential and intracellular calcium in myocyte cultures. AB - INTRODUCTION: Fast spatially resolved measurements of transmembrane potential (Vm) and intracellular calcium (Ca(i)2+) are important for studying mechanisms of arrhythmias and defibrillation. The goals of this work were (1) to develop an optical technique for simultaneous multisite optical recordings of Vm and Ca(i)2+, and (2) to determine the relationship between Vm and Ca(i)2+ during normal impulse propagation in myocyte cultures. METHODS AND RESULTS: Monolayers of neonatal rat myocytes were stained with fluorescent dye RH-237 (Vm) and Fluo 3AM (Ca(i)2+). Both dyes were excited at the same wavelength range. The emitted fluorescence was optically separated into components corresponding to changes in Vm and Ca(i)2+ and measured using two 16 x 16 photodiode arrays at a spatial resolution of up to 27.5 microm per diode and sampling rate of 2.5 kHz. The optical setup was adjusted so that there was no optical cross-talk between the two types of measurements, which was validated in experiments involving staining with either RH-237 or Fluo-3. The amplitude of Fluo-3 signals rapidly decreased during experiments due to dye leakage. Dye leakage was substantially reduced by application of 1 mM probenecid, a blocker of organic anion transport, which had no effect on action potential duration and only minor effect on conduction velocity. In double-stained preparations, during regular pacing Ca(i)2+ transients had a rise time of 14.2 +/- 2 msec, and they followed Vm upstrokes with a delay of 5.3 +/- 1 msec (n = 9). Durations of Vm and Ca(i)2+ transients determined at 50% level of signal recovery were 54.6 +/- 10 msec and 136 +/- 8 msec, respectively. Application of 2 microM nifedipine reduced the amplitude and duration of Ca(i)2+ transients without significantly affecting conduction velocity. CONCLUSION: The results demonstrate feasibility of simultaneous optical recordings of Vm and Ca(i)2+ transients with high spatial and temporal resolution. PMID- 10826935 TI - Maturational atrioventricular nodal physiology in the mouse. AB - INTRODUCTION: Dual AV nodal physiology is characterized by discontinuous conduction from the atrium to His bundle during programmed atrial extrastimulus testing (A2V2 conduction curves), AV nodal echo beats, and induction of AV nodal reentry tachycardia (AVNRT). The purpose of this study was to characterize in vivo murine maturational AV nodal conduction properties and determine the frequency of dual AV nodal physiology and inducible AVNRT. METHODS AND RESULTS: A complete transvenous in vivo electrophysiologic study was performed on 30 immature and 19 mature mice. Assessment of AV nodal conduction included (1) surface ECG and intracardiac atrial and ventricular electrograms; (2) decremental atrial pacing to the point of Wenckebach block and 2:1 conduction; and (3) programmed premature atrial extrastimuli to determine AV effective refractory periods (AVERP), construct A2V2 conduction curves, and attempt arrhythmia induction. The mean Wenckebach block interval was 73 +/- 12 msec, 2:1 block pacing cycle length was 61 +/- 11 msec, and mean AVERP100 was 54 +/- 11 msec. The frequency of dual AV nodal physiology increased with chronologic age, with discontinuous A2V2 conduction curves or AV nodal echo beats in 27% of young mice < 8 weeks and 58% in adult mice (P = 0.03). CONCLUSION: These data suggest that mice, similar to humans, have maturation of AV nodal physiology, but they do not have inducible AVNRT. Characterization of murine electrophysiology may be of value in studying genetically modified animals with AV conduction abnormalities. Furthermore, extrapolation to humans may help explain the relative rarity of AVNRT in the younger pediatric population. PMID- 10826936 TI - Effects of intracardiac radiofrequency current application on coronary artery vessels in young pigs. AB - INTRODUCTION: Radiofrequency current is widely used in children to ablate accessory AV pathways. Previous data in a pig model demonstrated coronary artery obstruction adjacent to radiofrequency current lesions 48 hours and 6 months after energy delivery. In the present study, the long-term effects (>6 months) of radiofrequency current application on coronary artery vessels in young pigs are assessed. METHODS AND RESULTS: Radiofrequency current (500 kHz) was delivered over 30 seconds in ten piglets (mean body weight 12.8 kg) using a steerable 6 French catheter with a 4-mm thermistor tip electrode (target temperature 75 degrees C). Lesions were created under fluoroscopic and electrocardiographic guidance at the lateral right atrial (RA) wall above the tricuspid valve orifice, and at the lateral left atrial and left ventricular wall adjacent to the mitral valve orifice. Selective coronary angiography and intravascular ultrasound (IVUS) studies were performed 3, 6, 9, and 12 months after energy application. After 12 months, the lesions were studied pathohistologically. All lesions consisted of compact fibrous tissue. RA lesions extended to the adjacent right coronary artery and led to coronary artery involvement with increased fibrous tissue in the adventitia and media and intimal thickening in three animals. Coronary arterial narrowing was documented by IVUS during follow-up in all three cases 9 months after energy application. Angiography failed to demonstrate coronary pathology in any of the three animals. CONCLUSION: The risk of late coronary artery lesions must be considered when catheter ablation at the RA wall is planned in children with free-wall accessory AV pathways. PMID- 10826937 TI - Bundle branch reentrant ventricular tachycardia with two distinct components of QRS complex due to simultaneous dual transseptal conduction. AB - We report the case of a patient with bundle branch reentrant ventricular tachycardia having two distinct components of the QRS complex due to simultaneous dual transseptal conduction. The macroreentrant circuit consisted of anterograde conduction down the left bundle branch and retrograde conduction up the right bundle branch. Extensively injured myocardium in the interventricular septum isolated the two separate transseptal passages, which exhibited a "fast" and "slow" conduction property, respectively. The QRS configuration resulted from a wavefront propagating from the left bundle branch via "fast" transseptal conduction, followed by another more delayed wavefront propagating via "slow" transseptal conduction, which played an important role in determining this unique QRS morphology. PMID- 10826938 TI - Focal ablation of chaotic atrial rhythm in an infant with cardiomyopathy. AB - Chaotic atrial rhythm in infants has been defined similar to multifocal atrial tachycardia in adults, implying a multifocal etiology. However, its ECG appearance resembles atrial fibrillation, which sometimes has a unifocal ectopic mechanism amenable to catheter ablation. Curative focal radiofrequency ablation was performed in a 4-month-old infant with chaotic atrial rhythm and dilated cardiomyopathy. Left ventricular function subsequently returned to normal. Reversibility of associated cardiomyopathy supports aggressive rhythm management of chaotic atrial rhythm. In this patient, the unifocal origin allows insight into the pathophysiology of the rhythm and demonstrates the potential utility of catheter ablation for refractory cases. PMID- 10826939 TI - Atrial tachycardia masquerading as atrial flutter following ablation of the subeustachian isthmus. AB - We report a case of atrial tachycardia masquerading as atrial flutter in a man who had previously undergone catheter ablation for atrial flutter. The recurrent arrhythmia was electrocardiographically almost identical to the prior atrial flutter; at repeat electrophysiologic study, although bidirectional conduction block was observed in the tricuspid annulus-inferior vena caval isthmus, the atrial arrhythmia was readily initiated. Activation mapping suggested typical atrial flutter, but entrainment techniques demonstrated intra-atrial reentry not involving the ablated isthmus. This case illustrates the need to apply entrainment techniques even in cases of apparent "typical" atrial flutter to confirm that putative ablation targets are necessary for tachycardia perpetuation. PMID- 10826940 TI - Left atrial fibrillation with regular right atrial activation and a single left to-right electrical interatrial connection: multisite mapping of dissimilar atrial rhythms. AB - We report two patients who had isolated atrial fibrillation in the left atrium and regular activation of the entire right atrium. Mapping of the arrhythmia was performed using a 64-electrode basket catheter that was inserted intravenously and deployed in the right and left atria. Both patients manifested a single, stable interatrial electrical connection conducting in a left-to-right direction, consistent with Bachmann's bundle location. The right and left sides of the interatrial septum were activated discordantly, each reflecting activation characteristics of the respective atria. A filtering effect at the level of interatrial septum was demonstrated by calculating the fibrillation intervals on both sides of the operative interatrial connection. It was concluded that differences in activation of the left and right surfaces of the interatrial septum and preferential use and the filtering effect of the interatrial connections play a significant role in explaining the differences in activation patterns of the left and right atria in patients with atrial fibrillation. PMID- 10826941 TI - Cardiac pacing in the long QT syndrome: review of available data and practical recommendations. AB - A review of published data on cardiac pacing in the long QT syndrome (LQTS) is presented, in the hope that optimization of patient selection and pacemaker programming will prevent arrhythmic death. LQT3 patients may derive particular benefit from pacing because the dispersion of repolarization worsens steeply during bradycardia in this genotype. However, concluding that other genotypes will not benefit from pacing is premature. Pacing may be especially beneficial for patients with "pause-dependent" arrhythmias. Programming should include a sufficiently fast lower rate limit. Features that allow heart rate slowing beyond the lower rate limit or that may trigger pauses must be programmed "off" because pauses are proarrhythmic in this population. Pause-prevention pacing algorithms may be beneficial. PMID- 10826942 TI - Cardiac arrhythmias: the genetic connection. PMID- 10826943 TI - Differential diagnosis of supraventricular tachycardia with concentric atrial activation. PMID- 10826944 TI - Virtual electrode polarization of ventricular epicardium during bipolar stimulation. PMID- 10826945 TI - Superantibodies: synergy of innate and acquired immunity. AB - The antibody molecule possesses a number of so-called unconventional binding sites in the variable domain that are expressed and function independently from the antigen-binding site. These sites are encoded in the germline, and are predominantly composed of framework residues. By this definition, these sites function as part of the innate immunity, and are not subject to antigen-driven mutation and maturation. In this article, we focus on the evidence for the function and utility of the self-binding domain. The self-binding or autophilic domain has been discovered on murine germline-encoded antibodies from the S107/T15 Vh family. Autophilic antibodies form self-complexes after attaching to targets, but remain monomeric in solution. A peptide has been identified that confers self-binding if chemically attached to antibodies. Because this modification enhances the overall avidity of antibodies for target binding, therapeutic and diagnostic antibodies can be biotechnologically improved. The concept of superantibodies is introduced here to describe the unique coexistence and synergism of acquired immunity with innate immunity via antigen-specific and unconventional functional domains. As not every antibody qualifies as a superantibody, biotechnological engineering can produce superantibodies with superior targeting and therapeutic properties. PMID- 10826946 TI - Tracking immunoglobulin variable-gene expression in HIV infection. AB - B-cell superantigens (SAgs) interact with normal human nonimmune immunoglobulins (Igs) independently of the light-chain isotype, and activate a large proportion of the B-cell repertoire. Recently, the major envelope protein of human immunodeficiency virus 1 (HIV-1), gp120, was found to exhibit SAg-like properties for B cells with potential pathological consequences for the infected host, including accelerated apoptosis and progressive loss of B cells. This unconventional mode of interaction contrasts with its binding to immunization induced antibodies, which requires the tertiary structure of the heavy- and light chain variable regions. Examining the temporal development of V(H)3+ antibodies in HIV-1-infected subjects over a 7-yr period showed that V(H)3+ antibodies specific for the gp120 SAg-binding site are deficient. Quantification of V(H)3+ antibodies, which impart protective responses to infectious agents, in serum samples from HIV-seropositive slow progressors and from patients who progressed to AIDS-related manifestations reveals that paucity in V(H)3+ antibodies is a marker of rapid clinical decline. Remarkably, anti-gp160 V(H)3+ antibodies show a gradual decrease in progressors and vary with time, depending on the viral load. Thus, V(H)3+ antibodies could play an important role in protection, and their underexpression may accelerate disease progression. Investigation of the structural basis of the interaction between human Igs and gp120 shows that the viral gp120 SAg can interact only with a subset of human V(H)3+ Igs. A number of amino acid-positions present primarily in the first and third framework regions of the Ig heavy-chain variable regions correlate with gp120 binding. These residues partially overlap with the Staphylococcus aureus protein A-binding site for V(H)3+ Igs. Overall, these interactions could represent a novel mechanism of humoral deficiency resulting from the capacity of a viral SAg to impact an important subset of the B-cell repertoire and to induce B-cell death by apoptosis. PMID- 10826948 TI - Diversification and selection mechanisms for the production of protein repertoires: lessons from the immune system. AB - The physiological mechanism for producing antigen-specific antibodies is based on a two-phase neo-Darwinian process: the first phase consists of diversity generation (formation of the repertoire), and the second phase is antigen mediated selection. In this article, we consider how the natural immunoglobulin gene-diversification processes can be exploited both in vivo and in vitro in order to allow the generation of novel antibody (and heterologous protein) repertoires. PMID- 10826947 TI - Epitope promiscuity of human monoclonal autoantibodies to T-cell receptor combining site determinants. AB - To characterize the binding specificity and light- and heavy-chain variable region usage in monoclonal human autoantibodies (mAAbs) to T-cell receptors, we constructed heterohybridomas from peripheral blood B cells of three rheumatoid arthritis (RA) patients. From a panel of more than 200 heterohybridomas secreting IgM autoantibodies binding to T-cell receptor Vbeta chain first complementarity determining segments (CDR1), we characterized two IgM/lambda molecules from a single patient in detail. These bound to both CDR1 peptide epitopes and intact TCR of recombinant single-chain T-cell receptor constructs, and to T-cell surface TCR. Spectratype analysis using epitopes mimicking a set of 24 Vbeta genes indicated that one molecule bound only a few members of the set, whereas the second showed considerable epitope promiscuity by binding to more than half of the tested CDR1 peptides. Both mAAbs used variants of a Vlambda3 gene that were very similar to one another and to the germline gene. The epitope-promiscuous autoantibody used a V(H)4 gene identical to a germline prototype, while the other incorporated a V(H)3 sequence differing in only a single residue from its germline prototype. The CDR3s of both were large and distinct from each other as well as from the corresponding segments of rheumatoid factors and "cold agglutinins" using the same or related V(H) germline genes. These mAAbs offer models for deciphering the basis of epitope promiscuity, and serve as candidates for direct use in immunomodulation because they are of intrinsic human origin and do not require molecular engineering to adapt them for use in therapy. PMID- 10826949 TI - Structural components responsible for peptide antigenicity. AB - The conformation of a peptide defines its antigenic specificity. In most cases, a free peptide is in a random form, whereas the same peptide within a protein has a defined conformation. Immunological cross-reactions between the two are rare. Nevertheless, there are cases where an anticonformational antibody may transconform (change the conformation of) the free peptide, allowing the reaction to occur. Based on such considerations, synthetic vaccines against infectious diseases may be constructed. The same is true for autoimmune diseases, where, at least in one case--that of the exacerbating-remitting type of multiple sclerosis- we have developed a polypeptidic drug-vaccine, copolymer 1 (denoted Copaxone), capable of suppressing the autoimmune phenomena in a specific way. This drug vaccine has been approved for treatment by the US Food and Drug Administration (FDA). PMID- 10826950 TI - Natural catalytic immunity is not restricted to autoantigenic substrates: identification of a human immunodeficiency virus gp 120-cleaving antibody light chain. AB - The autoimmune repertoire is well known from previous studies to be capable of producing catalytic antibodies directed to self-antigens. In the present study, we explored the ability of 26 monoclonal light chains (L chains) from multiple myeloma patients to cleave radiolabeled gp120, a foreign protein. One L chain with this activity was identified. 125I-gp120 and unlabeled gp120 were cleaved at several sites by the L chain, as shown by SDS-polyacrylamide gel electrophoresis, autoradiography, and immunoblotting, respectively. The apparent dissociation constant of the L chain was 130-145 nM, indicating high-affinity gp120 recognition. 125I-albumin was not cleaved by the L chain, and various proteins and peptides did not inhibit gp120 cleavage by the L chain, suggesting that the activity is not a nonspecific phenomenon. The substrate recognition determinants may be conserved in different HIV-1 strains, because gp120 isolated from strains SF2, MN, and IIIB was found to be cleaved by the L chain. Micromolar concentrations of a synthetic peptide corresponding to residues 23-30 of gp120 inhibited the cleavage of 125I-gp120, suggesting that these residues are components of the epitope recognized by the L chain. The toxic effect of gp120 in neuronal cultures was reduced by about 100-fold by pretreatment of the protein with the L chain. These observations open the possibility of utilizing gp120 cleaving antibodies in the treatment of AIDS. PMID- 10826951 TI - Does catalytic activity of Bence-Jones proteins contribute to the pathogenesis of multiple myeloma? AB - Some Bence-Jones proteins have been found to be capable of hydrolyzing DNA, chromogenic amide substrates, such as benzoylarginine p-nitroanilide, and natural oligopeptides, such as arginine vasopressin. Patients who excrete Bence-Jones protein with the DNA-nicking activity have shown moderately severe symptoms. When incubated with LLC-PK1 (porcine kidney proximal tubule) cells, some Bence Jones proteins penetrated the cytoplasm, and entered the nucleus with little or no degradation of epitopes. Intranuclear Bence Jones proteins ultimately induced DNA fragmentation in situ and cell death. This cytocidal activity was not directly associated with the DNA-nicking activity, since Bence Jones proteins with no detectable DNase activity also produced cell death. These results, however, suggest that the biological activities of Bence Jones proteins described here makes a significant contribution to the development and/or deterioration of multiple myeloma. PMID- 10826952 TI - DNA hydrolysis by monoclonal autoantibody BV 04-01. AB - Monoclonal anti-DNA autoantibody BV 04-01 catalyzed hydrolysis of DNA in the presence of Mg2+. Catalysis was associated with BV 04-01 IgG, Fab, and single chain-antibody (SCA) proteins. Cleavage of both ss and dsDNA was observed with efficient hydrolysis of the C-rich region of A7C7ATATAGCGCGT2, as well as a preference for cleaving within CG-rich regions of dsDNA. Data on specificity of ssDNA hydrolysis and kinetic data obtained from wild-type SCA, and two SCA mutants were used to model the catalytically active antibody site using the previously resolved X-ray structure of BV 04-01. The resulting model suggested that the target phosphodiester bond is activated by induction of conformational strain. In addition, the antibody-DNA complex contained a Mg2+ coordination site composed of the L32Tyr and L27dHis side chains and a DNA 3'-phosphodiester group. Induction of strain along with the metal coordination could be part of the mechanism by which this antibody catalyzes DNA hydrolysis. Sequence data for BV 04-01 V(H) and V(L) genes suggested that the proposed catalytic-antibody active site was germline-encoded. This observation suggests that catalytic activity might represent an important-rarely examined-function for some antibody molecules. PMID- 10826953 TI - Amidase and peptidase activities of polyclonal immunoglobulin G present in the sera of patients with rheumatoid arthritis. AB - Polyclonal Immunoglobulin (Ig) G from patients with rheumatoid arthritis (RA) and healthy subjects hydrolyzed carbobenzoxy-Val-Gly-Arg p-nitroanilide and D-Pro-Phe Arg p-nitroanilide. RA IgG exhibited higher activity against the former substrate, but not the latter. On the other hand, RA IgG showed reduced activity against D-Pro-Phe-Arg methylcoumarinamide, when compared with those of the healthy controls. These results suggest that RA IgGs differ from normal IgGs in the substrate specificity of amidase activity. Preliminary studies have shown that two out of three RA IgG samples cleaved a pentapeptide--Gln-Arg-Arg-Ala-Ala- which is assumed to be associated with the risk of developing RA (Gregersen, P. K. et al. (1987), Arthritis Rheum. 30, 1205-1213). By contrast, virtually no cleavage of the same peptide was observed with IgG from healthy controls. A peptide analog, Gln-Arg-Arg-Trp-Ala, was not cleaved at all by any IgGs examined either from RA patients or healthy controls. PMID- 10826954 TI - Secretory immunoglobulin A from healthy human mothers' milk catalyzes nucleic acid hydrolysis. AB - The human milk secretory immune system is the first line of protection for the newborn infant against various pathogens. Secretory IgA (sIgA), the typical immunoglobulin found in secretions, can fight infections through many mechanisms. Using different methods, we have shown that sIgA from the milk of healthy women possesses DNAse and RNAse activities. The catalytic center is localized in the light chain of catalytic sIgA, while the DNA-binding center is predominantly formed by its heavy chain. The enzymic properties and substrate specificity of catalytic sIgA distinguish it from other known DNases and RNases. It is reasonable to assume that the milk DNA- and RNA-hydrolyzing antibodies are capable not only of neutralizing viral and bacterial nucleic acids by binding these antigens as well as by hydrolyzing them. The DNA-hydrolyzing activity of Abs raises the possibility that these catalytic Abs may provide protective functions for the newborn through the hydrolysis of viral and bacterial nucleic acids. PMID- 10826956 TI - Idiotypic network dysregulation: a common etiopathogenesis of diverse autoimmune diseases. AB - The pathogenesis of autoimmune disease is still an enigma. Whereas the diverse clinical manifestations of many autoimmune diseases cannot be explained by the existence of autoantibodies, idiotypic dysregulation may provide an alternative explanation. Experimental models, serum level changes of pathogenic idiotypes during exacerbation and remission, and the increased expression of pathogenic idiotypes following common infections all support this notion. In this article we review experimental models of autoimmune disease induction (systemic lupus erythematosus, antiphospholipid syndrome, Goodpasture's syndrome, autoimmune thyroiditis, and vasculitis) by manipulation of the idiotypic network, and discuss the utilization of idiotypes for the immunotherapy of autoimmune diseases and other conditions that involve the immune system (e.g., atherosclerosis). PMID- 10826955 TI - Localization of a novel adhesion-promoting site on acetylcholinesterase using catalytic antiacetylcholinesterase antibodies displaying cholinesterase-like activity. AB - A monoclonal antibody (MAb) raised against human acetylcholinesterase was found to have catalytic activity. A similar phenomenon was observed in a polyclonal antibody raised against the same antigen. The antibodies were demonstrated to be pure, and no contamination with either acetylcholinesterase or butyrylcholinesterase was found. Both antibodies hydrolyzed acetylthiocholine, an acetylcholinesterase substrate, and the MAb followed Michaelis-Menten kinetics. Six other MAbs and one other polyclonal antibody showed no evidence of catalytic activity. Acetylcholinesterase is a key component in the transmission of the nerve impulse, and is also expressed nonsynaptically during embryonic development, and abnormalities in expression are seen in neural tumors and degenerative disorders. This unusual expression is believed to be associated with a novel function of the enzyme related to differentiation and cell adhesion. Autoantibodies to acetylcholinesterase have been observed in a variety of neurologic, muscular, and autoimmune disorders. In an investigation of the possible role of acetylcholinesterase in cell adhesion, we showed that the enzyme promoted neurite outgrowth in neuroblastoma cell lines, and conversely, that certain antiacetylcolinesterase antibodies abrogated cell-substrate adhesion. Interestingly, the antibodies most effective in this regard were catalytic. Preliminary epitope analysis indicated a conformational epitope in the N-terminal domain. This domain contains the active site within a deep gorge and the peripheral anionic site at the rim of the gorge. Peripheral-site inhibitors, but not active-site inhibitors, also interfered with adhesion, and competed with the catalytic monoclonal binding to acetylcholinesterase, indicating that the epitope recognized is associated with the peripheral anionic site. The inhibitor data also support the supposition that catalysis in these antibodies may have arisen from stable complexation of acetylcholinesterase with an inhibitor. We conclude that the catalytic antiacetylcholinesterase antibody interacts with structures associated with the peripheral anionic site, thus defining a novel site on the molecule involved in cell adhesion. This finding has implications for our understanding of the potential importance of this peripheral site in a variety of congenital, neoplastic, and degenerative conditions. PMID- 10826957 TI - Catalytic mechanism of an abzyme displaying a beta-lactamase-like activity. AB - A catalytic IgG (Ab2) displaying a beta-lactamase-like activity was previously obtained by using the antiidiotypic pathway: the particularity of this antibody is that it is a true antiidiotype of the beta-lactamase active site. We have previously demonstrated that this IgG has retained some of the structural information displayed by the beta-lactamase active site, evident from data that polyclonal anti-Ab2 antibodies (Ab3) recognize beta-lactamase. In this article, we investigated the catalytic mechanism of the abzyme compared to that of the enzyme. The experimental data allow us to hypothesize the catalytic residues required for catalysis. PMID- 10826958 TI - Nonspecific catalysis by protein surfaces. AB - Catalytic antibodies are the best available all-around enzyme mimics. They provide a unique experimental approach and some special insights into general questions about catalysis by enzymes. They offer enantiospecific reactions and levels of substrate binding that compare well with typical enzyme reactions, but not--so far--comparable catalytic efficiency. We and others have used the Kemp elimination as a probe of catalytic efficiency in antibodies. We compare these reactions with nonspecific catalysis by other proteins, and with catalysis by enzymes. Several simple reactions are catalyzed by the serum albumins with Michaelis-Menten kinetics, and can be shown to involve substrate binding and catalysis by local functional groups. Here we report the details of one investigation, which implicate known binding sites on the protein surface, and discuss implications for catalyst design and efficiency. PMID- 10826959 TI - Pyridoxal-5'-phosphate-dependent catalytic antibodies. AB - Cofactors--i.e., metal ions and coenzymes--extend the catalytic scope of enzymes and might have been among the first biological catalysts. They may be expected to efficiently extend the catalytic potential of antibodies. Monoclonal antibodies (MAbs) against Nalpha-phosphopyridoxyl-L-lysine were screened for 1) binding of 5'-phosphopyridoxyl amino acids, 2) binding of the planar Schiff base of pyridoxal-5'-phosphate (PLP) and amino acids, the first intermediate of all PLP dependent reactions, and 3) catalysis of the PLP-dependent alpha, beta elimination reaction with beta-chloro-D/L-alanine. Antibody 15A9 fulfilled all criteria and was also found to catalyze the cofactor-dependent transamination reaction of hydrophobic D-amino acids and oxo acids (k'cat = 0.42 min(-1) with D alanine at 25 degrees C). No other reactions with either D- or L-amino acids were detected. PLP markedly contributes to catalytic efficacy-it is a 10(4) times more efficient acceptor of the amino group than pyruvate. The antibody ensures reaction specificity, stereospecificity, and substrate specificity, and further accelerates the transamination reaction (k'cat(Ab)/k'cat(PLP) = 5 x 10(3)). The successive screening steps simulate the selection criteria that might have been operative in the evolution of protein-assisted pyridoxal catalysis. PMID- 10826960 TI - Catalytic antibodies for complex reactions: hapten design and the importance of screening for catalysis in the generation of catalytic antibodies for the NDA/CN reaction. AB - Success in generating catalytic antibodies as enzyme mimics lies in the strategic design of the transition-state analog (TSA) for the reaction of interest, and careful development of screening processes for the selection of antibodies that are catalysts. Typically, the choice of TSA structure is straightforward, and the criterion for selection in screening is often binding of the TSA to the antibody in a microtiter-plate assay. This article emphasizes the problems of TSA design in complex reactions and the importance of selecting antibodies on the basis of catalysis as well as binding to the TSA. The target reaction is the derivatization of primary amines with naphthalene-2,3-dicarboxaldehyde (NDA) in the presence of cyanide ion. The desired outcome is selective catalysis of formation of the fluorescent derivative in preference to nonfluorescent side products. In the study, TSA design was directed toward the reaction branch leading to the fluorescent product. Here, we describe a microtiter plate-based assay that is capable of detecting antibodies showing catalytic activity at an early stage. Of the antibodies selected, 36% showed no appreciable binding to any of the substrates tested, but did show catalytic activity in derivatizing one or more of the amino acids screened. In contrast, only two out of 77 clones that showed binding did not show catalysis. Thus, in this complex system, observation of binding is a good predictor of the presence of catalytic activity, and failure to observe binding is a poor predictor of the absence of catalytic activity. PMID- 10826961 TI - How and why 41S-2 antibody subunits acquire the ability to catalyze decomposition of the conserved sequence of gp41 of HIV-1. AB - It has become well known that antibodies obtained by immunization with the ground state of peptides can display proteolytic activity. Our antibody light chain produced by immunization with the peptide RGPDRPEGIEEEGGERDRD, a highly conserved sequence in envelope gp41 of HIV-1 showed the ability to cleave this peptide. Moreover, its heavy chain also decomposed the peptide, although this occurred at lower activity levels compared with the light chain, while the whole antibody did not show any catalytic activity. From molecular modeling, the light and heavy chains of the antibody were deduced to possess catalytic triads (Asp, His, and Ser) in their steric conformations, which may be responsible for the observed proteolytic activity. PMID- 10826962 TI - Nucleophilic proteolytic antibodies. AB - Proteolytic antibodies appear to utilize catalytic mechanisms akin to nonantibody serine proteases, assessed from mutagenesis and protease-inhibitor studies. The catalytic efficiency derives substantially from the ability to recognize the ground state with high affinity. Because the proteolytic activity is germline encoded, catalysts with specificity for virtually any target polypeptide could potentially be developed by applying appropriate immunogens and selection strategies. Analysis of transition-state stabilizing interactions suggests that chemical reactivity of active-site serine residues is an important contributor to catalysis. A prototype antigen analog capable of reacting covalently with nucleophilic serine residues permitted enrichment of the catalysts from a phage displayed lupus light-chain library. Further mechanistic developments in understanding proteolytic antibodies may lead to the isolation of catalysts suitable for passive immunotherapy of major diseases, and elicitation of catalytic immunity as a component of prophylactic vaccination. PMID- 10826963 TI - Inhibition and labeling of enzymes and abzymes by phosphonate diesters. AB - Reactive phosphonate diesters were designed and prepared as inhibitors of serine proteases and esterases. Inactivation of trypsin, chymotrypsin, and butyrylcholinesterase was determined by residual enzymatic activity as well as by the release of a chromogenic or fluorogenic product of the inhibition reaction. Second-order rate constants were determined from rates of nitrophenol formation. Application of the reaction for active-site titration of enzyme preparations is demonstrated. A basic functional group present in the nitrophenyl tropane phosphonate diester was shown to confer selectivity for inactivation of trypsin and chymotrypsin. Biotinylated derivatives of the phosphonate diesters were prepared to permit analysis of proteins modified in the inhibition reaction. Labeled polypeptides were resolved by SDS-PAGE, electroblotted, and detected by streptavidin-peroxidase staining. A detection limit of less than 4 ng, corresponding to 20 nM of trypsin, was demonstrated. Pretreatment of enzymes with DFP or nonbiotinylated phosphonates specifically blocks the labeling. This technique permits identification of serine proteases in complex mixtures with good sensitivity and specificity. PMID- 10826964 TI - Autoantibodies to thyroglobulin in health and disease. AB - Thyroglobulin (Tg)--a heavily glycosylated, iodinated protein--is a major autoantigen in autoimmune thyroiditis. Tg also induces thyroiditis by immunization of experimental animals. Humans with chronic lymphocytic thyroiditis characteristically produce autoantibodies to thyroglobulin, but similar autoantibodies are also found in some clinically normal, euthyroid individuals. A comparison of the fine specificity of autoantibodies in humans and in experimentally immunized mice was carried out, based on their ability to inhibit a panel of monoclonal antibodies (MAbs). Patients with autoimmune thyroid disease, as well as normal individuals, produced autoantibodies mainly to the conserved, cross-reactive determinants of thyroglobulin. Patients developed additional autoantibodies to species-restricted epitopes. The determinants recognized by patients with Graves' disease differed in some respects from epitopes recognized by thyroiditis patients or patients with differentiated thyroid carcinoma. Similarly, mice that are genetically susceptible to thyroiditis produced autoantibodies that reacted with the mouse-specific antigenic determinants. Using an autoantibody that reacts with one of the epitopes associated with thyroiditis, a reactive 15-kDa fragment of human Tg- localized at the carboxy end of the molecule--was isolated and sequenced. Iodine plays an important role in the precise specificity of the disease-associated epitope, since T cells from patients with thyroiditis react with iodinated but not noniodinated human thyroglobulin. Addition of iodine to Tg generates new or exposes cryptic epitopes. Use of a selected MAb as a surrogate for the T-cell receptor suggests that a specific iodine-containing epitope is sometimes involved in recognition. Finally, thyroglobulin-reactive autoantibodies exhibit proteolytic activity on thyroglobulin. PMID- 10826965 TI - Autoantibodies to nuclear antigens: correlation between cytotoxicity and DNA hydrolyzing activity. AB - The cytotoxicity of DNA-specific autoantibodies from sera of patients with systemic lupus erythematosis (SLE) and with lymphoproliferative diseases, and from blood of healthy donors was examined on tumor-cell lines L929 and HL-60. DNA binding IgG fractions from SLE and chronic lymphocytic leukemia (CLL) sera were cytotoxic at concentrations of up to 10(-10) M. No detectable changes in cell viability were observed after incubation with antibodies devoid of DNA-binding activity and DNA-specific antibodies isolated from blood of healthy donors and patients with T-cell lymphoma, B-cell lymphosarcoma, and acute B-cell leukemia. There was good correlation between the cytotoxic activity and DNA-hydrolyzing activity of anti-DNA antibodies. The cytotoxic effect of DNA-binding antibodies presumably was complement-independent, because it was attributed only to the Fab fragment. The cytotoxic effect was completely inhibited by preincubation with double-stranded DNA (dsDNA). Both the cytotoxic effect and the DNA-hydrolyzing activity of anti-DNA antibodies were significantly increased in the antibody fraction that displayed cross-reactivity with nuclear matrix proteins. Possible mechanisms for the formation and pathogenicity of cytotoxic anti-DNA antibodies are discussed in this article. PMID- 10826966 TI - Multifunctional antigens of A. fumigatus and specific antibodies. AB - The majority of Aspergillus-induced infections in man are caused by the pathogenic fungus A. fumigatus, which secretes biologically and immunologically active glycosylated and nonglycosylated proteins. The complexity in the antigenic structure of A. fumigatus and the varying host immune responses lead to a wide spectrum of clinical conditions such as allergic bronchopulmonary aspergillosis (ABPA), aspergilloma, and invasive aspergillosis. It is reported that 15-20% of allergic asthmatics suffer from Aspergillus-induced allergies. The incidence of opportunistic infections, including Aspergillus infections, has risen because of the increase in the incidence of HIV and tuberculosis. Allergic bronchopulmonary aspergillosis is an immunologically significant clinical form where type I and type III hypersensitivity reactions are involved in pathogenesis. High levels of specific IgE and IgG antibodies in these patients are of diagnostic value. Molecular characterization of certain immunodominant allergens and antigens of A. fumigatus revealed the presence of complex carbohydrate moieties, heat-shock proteins, enzyme activities such as elastase, protease, catalase, dismutase, and cytotoxic ribonuclease. A Con A binding allergen/antigen (45 kDa) and Con A nonbinding allergen/antigen (18 kDa, Asp fI) have a multifunctional nature. The multifunctional nature of these antigens may play an important role in the pathogenesis of the disease. Significant amounts of a major allergen/antigen of molecular weight 18 kDa is excreted in large amounts through the urine of patients with invasive aspergillosis. Studies on the structure-function relationship of the 18-kDa allergen/antigen revealed the involvement of tryptophan residues in binding with monoclonal antibodies (MAbs). Also, the histidine residues and cysteine disulfide bonds of the 18-kDa allergen are involved in its catalytic activity. The high load of multifunctional antigens in the serum of patients for prolonged periods, the presence of high levels of specific antibodies, and the absence of protective antibodies in ABPA patients have necessitated studies on the functional properties of the antibodies. The present study shows significant immunoreactivity of antibodies in patients of ABPA to fibronectin and collagen. Analysis of IgG antibodies from the patients of ABPA showed the presence of DNA-cleaving activity. These observations offer a new line of thinking in understanding the mechanism of pathogenesis of Aspergillus induced clinical manifestations, and may lead to novel approaches to intervention in the inflammation and infection caused by fungal pathogens. PMID- 10826967 TI - A preliminary study for isolation of catalytic antibodies by histidine ligand affinity chromatography as an alternative to conventional protein A/G methods. AB - Catalytic autoimmune antibodies from the sera of lupus patients were purified using histidyl-aminohexyl-Sepharose gel and compared with the antibodies purified with protein A and protein G affinity chromatography. The IgG preparations from the histidine affinity column had a much higher catalytic activity in hydrolyzing the peptide substrate Pro-Phe-Arg-methylcoumarinamide compared to the antibodies obtained by the conventional protein A/G method. This preservation of catalytic activity is attributed to the gentle buffer conditions used in the histidine ligand method that allowed the integrity of three-dimensional structure of purified catalytic antibodies. Thus, histidine affinity offer a superior method for isolating autoimmune catalytic antibodies. PMID- 10826968 TI - Recent developments in the encoding and deconvolution of combinatorial libraries. AB - The value of molecular libraries generated by combinatorial methods is largely dependent on the ease and ability to deconvolute or decode the structure of compounds of interest after screening the library. Following the introduction of promising concepts in the early 1990s, there has been considerable progress in the development and refinement of methodologies to address this issue. PMID- 10826969 TI - Evolution of DNA and RNA as catalysts for chemical reactions. AB - In vitro selection from combinatorial nucleic acid libraries has provided new RNA and DNA molecules that have catalytic properties. Catalyzed reactions now go far beyond self-modifying reactions of nucleic acid molecules. The future application of in vitro selected RNA and DNA catalysts in bioorganic synthesis appears promising. PMID- 10826970 TI - Evolutionary optimisation of enzymes. AB - Aside from the demonstration that individual molecular traits of enzymes can be evolutionarily optimised, the discovery that several traits can be simultaneously optimised is a major advance. The first observations of the effects of evolutionary optimisation at the structural level, through X-ray crystallography, reinforce the view that enzymes are best optimised by evolution and not by design. PMID- 10826971 TI - Optical detection methods for combinatorial libraries. AB - The main interests in the development of new combinatorial assays are the reduction of time for screening and an increase in the number of samples measured in parallel. The variety of detection methods is increasing, but the optimal one has not yet been determined. In the past two years, the first parallel detection methods for non-labelled compounds have been developed. PMID- 10826972 TI - Structure-based library design: molecular modelling merges with combinatorial chemistry. AB - Recent advances in both computational and experimental techniques now allow a very fruitful interplay of computational and combinatorial chemistry in the structure-based design of combinatorial libraries. PMID- 10826973 TI - Molecular evolution: dynamic combinatorial libraries, autocatalytic networks and the quest for molecular function. AB - The principles of Darwinian evolution have been explored in molecular systems such as autocatalytic networks and dynamic combinatorial libraries. Molecular evolution in such systems manifests itself as ligand or receptor amplification by selection. Research efforts exploring these concepts may provide a mechanism for the identification of novel catalysts, molecular receptors and bioactive molecules. PMID- 10826974 TI - Functional diversity of compound libraries. AB - The ideal designed screening library should contain compounds with a variety of structural shapes and molecular properties, while avoiding redundancies. Other requirements involve the need to find structurally distinct leads and to recognise drug-like molecules. Functional diversity analysis is one way in which these objectives can be achieved. For this, molecular descriptions that relate to both structure and properties of molecules are needed, as well as their evaluation in terms of biological relevance. PMID- 10826975 TI - Recent applications of polymer-supported reagents and scavengers in combinatorial, parallel, or multistep synthesis. AB - There has been a wealth of recent reports concerning support-bound reagents and scavengers in the solution-phase synthesis of compound libraries and natural products. Important advances in 1999 include the continued development and use of novel reagents for heterocycle synthesis, the increased use of catch-and-release purification, and the development of increasingly sophisticated techniques to allow sequestering of many types of impurities from desired compounds. These techniques have all been combined to enable the complicated multistep synthesis of natural products and of libraries of novel drug-like molecules, without conventional purification. PMID- 10826976 TI - The discovery of new isocyanide-based multi-component reactions. AB - Multi-component reactions are finding increasing use in the discovery process of new drugs and agrochemicals. Some years ago they were considered as highly exotic types of organic reactions. Recently, many groups have realized that the field of multi-component reactions is full of new opportunities. These comprise fast and efficient production of small-molecular-weight compound libraries, highly atom economic chemistry, very large chemical libraries, improvements in total synthesis, and applications in bioconjugate chemistry, as well as in related fields (e.g. chemical biology). PMID- 10826977 TI - Optimization of chemical libraries by neural networks. AB - Neural networks are finding ever-more applications in the design of combinatorial libraries. These can be divided into two types: Kohonen (self-organizing) maps, and feed-forward networks. While the number of applications is currently quite limited, a rapid increase in publications in this area can be expected in the next few years from the rapid development of general combinatorial chemistry technology. PMID- 10826978 TI - Immobilized catalysts in combinatorial chemistry. AB - As a new methodology for library synthesis in combinatorial chemistry, the use of immobilized catalysts and multi-component reactions is focused. In the past two years, many advances have been made in this emerging field, leading to the efficient library synthesis of, for example, quinolines, amino ketones and amino esters. PMID- 10826979 TI - High-diversity combinatorial libraries. AB - The synthesis of complex chemical structures by combinatorial chemistry has gained considerable interest. New chemical methods have been developed that enable the synthesis of compound libraries exhibiting structural diversities similar to those of natural products. The concept of 'chemical genomics' has been introduced, reflecting a new quality of understanding and creating the relationship between diverse artificial chemical structures with the space of biological responses and possible protein ligands. PMID- 10826980 TI - Lantibiotics and microcins: polypeptides with unusual chemical diversity. AB - Bacterial-derived antimicrobial polypeptides enjoy a large degree of structural and chemical diversity. Two well-studied examples of such polypeptides are the lanthionine-containing lantibiotics produced by a variety of Gram-positive bacteria, and their Gram-negative counterparts, the microcins. Both groups are produced as gene-encoded precursor peptides and undergo post-translational modification to generate the active moieties. Structure elucidation of novel lantibiotics and microcins has recently uncovered further novel structural and chemical features and, combined with the generation of analogue peptides by genetic manipulation, new insights into structure-function relationships have been gained. Furthermore, study of the mode of action of the lantibiotics nisin and mersacidin has revealed their use of a 'docking molecule' in the target cell to facilitate their biological activities. Meanwhile, in vitro studies with microcin B17 have helped to uncover the molecular mechanisms by which post translational modification results in the formation of heterocyclic oxazole and thiazole rings. From a practical standpoint, both groups of polypeptides represent new lead structures for future development of antimicrobial agents, whilst the identification of the 'docking molecules' represents a step forward in the search for novel targets for future antibiosis. PMID- 10826981 TI - Synthesis of natural-product-based compound libraries. AB - Natural products cover a diversity space not yet available from synthetic libraries, with an unrivalled success rate as drug leads. The combinatorial synthesis of non-oligomeric natural-product-based libraries, however, is still limited to few examples because access to easily modified units strongly depends on the availability of a core structure either from a natural source, or through a suitable synthetic route. Only a few resourceful groups have managed the latter approach for more demanding multifunctional natural drug leads, such as epothilones. PMID- 10826982 TI - Zebrafish: a model system for the study of human disease. AB - The zebrafish (Danio rerio) is a powerful model organism for the study of vertebrate biology, being well suited to both developmental and genetic analysis. Large-scale genetic screens have identified hundreds of mutant phenotypes, many of which resemble human clinical disorders. The creation of critical genetic reagents, coupled with the rapid progress of the zebrafish genome initiative directed by the National Institutes of Health, are bringing this model system to its full potential for the study of vertebrate biology, physiology and human disease. PMID- 10826983 TI - Strategies in complex disease mapping. AB - Dissecting the genetics of common, complex disorders remains one of the great challenges in human genetics. The acceleration of human genome sequence determination, improvements in informatics, large-scale identification of single nucleotide polymorphisms and improvements in scoring technologies have now increased the feasibility of identifying polymorphisms that predispose to common disease. PMID- 10826984 TI - Potential mechanisms of mutations that affect neuronal migration in man and mouse. AB - Mutations in the genes that encode filamin-1, Lis1 and doublecortin are responsible for X-linked lissencephaly in man, whereas mutations in the genes that encode Cdk5, its activator p35 and the reelin-signaling pathway disturb migration and architectonic development in mice. To understand the action of genes that control neuronal migration and the phenotype of corresponding defects, it might be as important to consider the positioning of the nucleus as it is to consider the guidance of the leading process. PMID- 10826985 TI - Toward quantitative genetic analysis of host and parasite traits in the manifestations of Plasmodium falciparum malaria. AB - The complex human and parasite determinants that influence disease severity in Plasmodium falciparum malaria reflect thousands of years of selective pressure. Emerging genetic and genomic resources offer the prospect of unraveling interactions of these determinants. PMID- 10826986 TI - Inherited disorders of transport in the liver. AB - There has been a recent explosion in our understanding of hepatic transport processes. Much of this has resulted from the investigation of human diseases involving the liver and the use of animal models. The physiological roles of many of these transporters have been well characterised previously but have, until now, been resistant to molecular cloning. PMID- 10826987 TI - Ion channels and epilepsy in man and mouse. AB - Inherited disorders of voltage-gated ion channels are a recently recognized etiology of epilepsy in the developing and mature central nervous system. Two human epilepsy syndromes, benign familial neonatal convulsions and generalized epilepsy with febrile seizures plus, represent K+ and Na+ channelopathies, and other newly defined syndromes have now been mapped to chromosomal regions that are rich in ion channel genes. Experimental mouse models promise a resolution of their intriguing pathophysiology, which includes a diverse array of cellular phenotypes consistent with the differential contributions of individual channels to excitability in neural networks. PMID- 10826988 TI - X-linked non-specific mental retardation. AB - Non-specific mental retardation is a very common and genetically heterogeneous disorder but, to date, only six genes related to this condition have been identified. Five of these six have been found in the past two years, through positional-cloning efforts of mapped X-linked families. The characteristics of the newly identified genes are providing insights into the molecular mechanisms of mental impairment and the development of cognitive functions. PMID- 10826989 TI - Genetics of hypertension. AB - In the past year, substantial progress has been made in both mapping and fine mapping the genes involved in blood pressure regulation. Genome scans have been carried out in humans and mice and these reveal many new potential chromosomal locations for blood pressure susceptibility loci. The chromosomal regions containing blood pressure genes for many of the inbred hypertensive rat models have been refined using new congenic strains. Further genetic studies support a role for antiotensinogen, aldosterone synthase and a region close to the epithelial sodium channel in blood pressure regulation. Finally, comprehensive single-nucleotide polymorphism analysis of cardiovascular genes has been undertaken using chip technology. PMID- 10826990 TI - The genetics of Parkinson's disease. AB - The effort to map the entire human genome has led recently to the important milestone publication in late 1999 of the complete sequence of chromosome 22. This has been facilitated by increasingly sophisticated tools for genetic analysis and the ensuing wealth of detailed genetic information. The quest for genetic factors contributing to Parkinson's disease and parkinsonian disorders has revealed a progressively complex picture implicating gene mutations in the rarer, autosomally inherited forms of Parkinson's disease and the interplay of genetic and/or environmental factors in the common sporadic forms of the disorder. These findings not only reiterate the complex genetic heterogeneity of Parkinson's disease but could also point towards common pathogenic mechanisms in Parkinson's disease and related neurodegenerative disorders. PMID- 10826991 TI - Methyl-CpG-binding protein 2 mutations in Rett syndrome. AB - The X-linked methyl-CpG-binding protein 2 gene (MECP2) encodes a protein that links DNA methylation to transcriptional repression mediated by histone deacetylases. Mutations in MECP2 have been found in 76% of classic Rett syndrome patients. Favourable nonrandom X chromosome inactivation ameliorates the phenotype. PMID- 10826992 TI - Genetics of ventral forebrain development and holoprosencephaly. AB - The disease holoprosencephaly is the basis of the most common structural anomaly of the developing forebrain in humans. Numerous teratogens when administered during early gastrulation, have been associated with this condition. Recent studies have characterized molecules expressed in the prechordal plate which are critical for normal brain formation. Perturbation of signaling pathways involving these molecules have been shown to cause holoprosencephaly in humans and other organisms. PMID- 10826993 TI - Left-right axis malformations in man and mouse. AB - The study of left-right axis malformations in man and mouse has greatly advanced understanding of the mechanisms regulating vertebrate left-right axis formation. Recently, the roles of the TGF-beta family, Sonic hedgehog and fibroblast growth factor signaling, homeobox genes, and cilia in left-right axis determination have been more clearly defined. The identification of genes and environmental factors affecting left-right axis formation has important implications for understanding human laterality defects. PMID- 10826994 TI - Genetic susceptibility to malaria getting complex. AB - The evolution of sickle cell disease illustrates the powerful selective pressure of malaria in Africa, and candidate gene association studies have identified more than ten putative susceptibility determinants involving erythrocytes or the immune system. Efforts at present are aimed at understanding the functional basis of known associations, and at developing both linkage- and association-based approaches of genome-wide screening for novel susceptibility factors. PMID- 10826995 TI - Genetics of the neuronal ceroid lipofuscinoses. AB - The neuronal ceroid lipofuscinoses (NCLs) are an intriguing group of inherited neurodegenerative disorders characterized by blindness, progressive psychomotor deterioration and death of neocortical neurons. Clinically, four major NCL groups have been identified: infantile, late infantile, juvenile and adult. In recent years, our understanding of the molecular basis of different NCLs has advanced significantly. The accumulation of autofluorescent material in patients' tissues has been shown to be caused by defects in either lysosomal enzymes or in novel membrane proteins of unknown function. Although the accumulated material is biochemically well defined and some of the causative mutations are known, a unifying hypothesis for the molecular basis of the NCLs remains elusive. Further work will be required to characterize the interactiving molecules and metabolic pathways involved in the pathogenesis of NCLs. PMID- 10826996 TI - Inherited disorders of ion transport in the intestine. AB - Within the past year, it has been established that the gene mutated in the human disorder congenital chloride diarrhea encodes a major Cl-/HCO3- exchanger at the apical membrane of gut epithelial cells. A major apical Na+/H+ exchanger has also been identified. New insight into metal ion absorption has been gained, and several new transporters without cognate diseases have been cloned. PMID- 10826997 TI - Insulin-like growth factor (IGF)-binding proteins: interactions with IGFs and intrinsic bioactivities. AB - The insulin-like growth factor (IGF)-binding proteins (IGFBPs) are a family of six homologous proteins with high binding affinity for IGF-I and IGF-II. Information from NMR and mutagenesis studies is advancing knowledge of the key residues involved in these interactions. IGF binding may be modulated by IGFBP modifications, such as phosphorylation and proteolysis, and by cell or matrix association of the IGFBPs. All six IGFBPs have been shown to inhibit IGF action, but stimulatory effects have also been established for IGFBP-1, -3, and -5. These generally involve a decrease in IGFBP affinity and may require cell association of the IGFBP, but precise mechanisms are unknown. The same three IGFBPs have well established effects that are independent of type I IGF receptor signaling. IGFBP 1 exerts these effects by signaling through alpha(5)beta(1)-integrin, whereas IGFBP-3 and -5 may have specific cell-surface receptors with serine kinase activity. The regulation of cell sensitivity to inhibitory IGFBP signaling may play a role in the growth control of malignant cells. PMID- 10826998 TI - Effects of high-fat diet and exercise training on intracellular glucose metabolism in rats. AB - We examined the effects of high-fat diet (HFD) and exercise training on insulin stimulated whole body glucose fluxes and several key steps of glucose metabolism in skeletal muscle. Rats were maintained for 3 wk on either low-fat (LFD) or high fat diet with or without exercise training (swimming for 3 h per day). After the 3-wk diet/exercise treatments, animals underwent hyperinsulinemic euglycemic clamp experiments for measurements of insulin-stimulated whole body glucose fluxes. In addition, muscle samples were taken at the end of the clamps for measurements of glucose 6-phosphate (G-6-P) and GLUT-4 protein contents, hexokinase, and glycogen synthase (GS) activities. Insulin-stimulated glucose uptake was decreased by HFD and increased by exercise training (P < 0.01 for both). The opposite effects of HFD and exercise training on insulin-stimulated glucose uptake were associated with similar increases in muscle G-6-P levels (P < 0.05 for both). However, the increase in G-6-P level was accompanied by decreased GS activity without changes in GLUT-4 protein content and hexokinase activities in the HFD group. In contrast, the increase in G-6-P level in the exercise trained group was accompanied by increased GLUT-4 protein content and hexokinase II (cytosolic) and GS activities. These results suggest that HFD and exercise training affect insulin sensitivity by acting predominantly on different steps of intracellular glucose metabolism. High-fat feeding appears to induce insulin resistance by affecting predominantly steps distal to G-6-P (e.g., glycolysis and glycogen synthesis). Exercise training affected multiple steps of glucose metabolism both proximal and distal to G-6-P. However, increased muscle G-6-P levels in the face of increased glucose metabolic fluxes suggest that the effect of exercise training is quantitatively more prominent on the steps proximal to G 6-P (i.e., glucose transport and phosphorylation). PMID- 10826999 TI - Ability of insulin to modulate hepatic glucose production in aging rats is impaired by fat accumulation. AB - Increased total fat mass (FM) and visceral fat (VF) may account in part for age associated decrease in hepatic insulin action. This study determined whether preventing the changes in body fat distribution abolished this defect throughout aging. We studied the F(1) hybrid of Brown Norway-Fischer 344 rats (n = 29), which we assigned to caloric restriction (CR) or fed ad libitum (AL). CR (55% of the calories consumed by AL) was initiated and used at 2 mo to prevent age dependent increases in FM and VF. AL rats were studied at 2, 8, and 20 mo; CR rats were studied at 8 and 20 mo. VF and FM remained unchanged throughout aging in CR rats. AL-fed rats at 8 and 20 mo had over fourfold higher FM and VF compared with both CR groups. Insulin clamp studies (3 mU. kg(-1). min(-1) with somatostatin) were performed to assess hepatic insulin sensitivity. Prevention of fat accretion resulted in a marked improvement in insulin action in the suppression of hepatic glucose production (HGP) (6.3 +/- 0.3 and 7.2 +/- 1.2 mg. kg(-1). min(-1) in 8- and 20-mo CR rats vs. 8.3 +/- 0.5 and 10.8 +/- 0.9 mg. kg( 1). min(-1) in 8- and 20-mo AL rats, respectively). The rate of gluconeogenesis (by enrichment of hepatic uridine diphosphate glucose and phosphoenolpyruvate pools by [(14)C]lactate) was unchanged in all groups. The improvement in hepatic insulin action in the CR group was mostly due to effective suppression of glycogenolysis (4.4 +/- 0.3 and 4.9 +/- 0.3 mg. kg(-1). min(-1) in 8- and 20-mo CR rats vs. 5.8 +/- 0.6 and 8.2 +/- 1.0 mg. kg(-1). min(-1) in 8- and 20-mo AL rats, respectively). The results demonstrated the preservation of hepatic insulin action in aging CR rats. Therefore, body fat and its distribution are major determinants of age-associated hepatic insulin resistance. PMID- 10827000 TI - Regulation of MAP kinase pathway activity in vivo in human skeletal muscle. AB - Insulin and exercise potently stimulate glucose metabolism and gene transcription in vivo in skeletal muscle. A single bout of exercise increases the rate of insulin-stimulated glucose uptake and metabolism in skeletal muscle in the postexercise period. The nature of the intracellular signaling mechanisms that control responses to exercise is not known. In mammalian tissues, numerous reports have established the existence of the mitogen-activated protein (MAP) kinase signaling pathway that is activated by a variety of growth factors and hormones. This study was undertaken to determine how a single bout of exercise and physiological hyperinsulinemia activate the MAP kinase pathway. The euglycemic-hyperinsulinemic clamp and cycle ergometer exercise techniques combined with percutaneous muscle biopsies were used to answer this question. In healthy subjects, within 30 min, insulin significantly increased MAP kinase [isoforms p42(MAPK) and p44(MAPK) (ERK1 and ERK2)] phosphorylation (141 +/- 2%, P < 0.05) and activity (177 +/- 5%, P < 0.05), and the activity of its upstream activator MEK1 (161 +/- 16%, P < 0.05). Insulin also increased the activity of the MAP kinase downstream substrate, the p90 ribosomal S6 kinase 2 (RSK2) almost twofold (198 +/- 45%, P < 0.05). In contrast, a single 30-min bout of moderate intensity exercise had no effect on the MAP kinase pathway activation from MEK to RSK2 in muscle of healthy subjects. However, 60 min of exercise did increase extracellular signal-related kinase activity. Therefore, despite similar effects on glucose metabolism after 30 min, insulin and exercise regulate the MAP kinase pathway differently. Insulin more rapidly activates the MAP kinase pathway. PMID- 10827001 TI - Kinetics of L-[1-(13)C]leucine when ingested with free amino acids, unlabeled or intrinsically labeled casein. AB - In two groups of five adults, each adapted to two different dietary regimens for 6 days, the metabolic fate of dietary [1-(13)C]leucine was examined when ingested either together with a mixture of free amino acids simulating casein (extrinsically labeled; condition A), along with the intact casein (extrinsically labeled; condition B), or bound to casein (intrinsically labeled; condition C). Fed state leucine oxidation (Ox), nonoxidative leucine disposal (NOLD), protein breakdown, and splanchnic uptake have been compared using an 8-h oral [1 (13)C]leucine and intravenous [(2)H(3)]leucine tracer protocol while giving eight equal hourly mixed meals. Lower leucine Ox, increased NOLD, and net protein synthesis were found with condition C compared with condition A (19.3 vs. 24.9; 77 vs. 55.8; 18.9 vs. 12.3 micromol. kg(-1). 30 min(-1); P < 0.05). Ox and NOLD did not differ between conditions B and C. Splanchnic leucine uptake calculated from [1-(13)C]- and [(2)H(3)]leucine plasma enrichments was between 24 and 35%. These findings indicate that the form in which leucine is consumed affects its immediate metabolic fate and retention by the body; the implications of these findings for the tracer balance technique and estimation of amino acid requirements are discussed. PMID- 10827002 TI - Somatostatin restrains the secretion of glucagon-like peptide-1 and -2 from isolated perfused porcine ileum. AB - Suspecting that paracrine inhibition might influence neuronal regulation of the endocrine L cells, we studied the role of somatostatin (SS) in the regulation of the secretion of the proglucagon-derived hormones glucagon-like peptide-1 and -2 (GLP-1 and GLP-2). This was examined using the isolated perfused porcine ileum stimulated with acetylcholine (ACh, 10(-6) M), neuromedin C (NC, 10(-8) M), and electrical nerve stimulation (NS) with or without alpha-adrenergic blockade (phentolamine 10(-5) M), and perfusion with a high-affinity monoclonal antibody against SS. ACh and NC significantly increased GLP secretion, whereas NS had little effect. SS immunoneutralization increased GLP secretion eight- to ninefold but had little influence on the GLP responses to ACh, NC, and NS. Basal SS secretion (mainly SS28) was unaffected by NS alone. Phentolamine + NS and NC abstract strongly stimulated release mainly of SS14, whereas ACh had little effect. Infused intravascularly, SS14 weakly and SS28 strongly inhibited GLP secretion. We conclude that GLP secretion is tonically inhibited by a local release of SS28 from epithelial paracrine cells, whereas SS14, supposedly derived from enteric neurons, only weakly influences GLP secretion. PMID- 10827003 TI - Insulin-mediated translocation of GLUT-4-containing vesicles is preserved in denervated muscles. AB - Skeletal muscle denervation decreases insulin-sensitive glucose uptake into this tissue as a result of marked GLUT-4 protein downregulation ( approximately 20% of controls). The process of insulin-stimulated glucose transport in muscle requires the movement or translocation of intracellular GLUT-4-rich vesicles to the cell surface, and it is accompanied by the translocation of several additional vesicular cargo proteins. Thus examining GLUT-4 translocation in muscles from denervated animals allows us to determine whether the loss of a major cargo protein, GLUT-4, affects the insulin-dependent behavior of the remaining cargo proteins. We find no difference, control vs. denervated, in the insulin-dependent translocation of the insulin-responsive aminopeptidase (IRAP) and the receptors for transferrin and insulin-like growth factor II/mannose 6-phosphate, proteins that completely (IRAP) or partially co-localize with GLUT-4. We conclude that 1) denervation of skeletal muscle does not block the specific branch of insulin signaling pathway that connects receptor proximal events to intracellular GLUT-4 vesicles, and 2) normal levels of GLUT-4 protein are not necessary for the structural organization and insulin-sensitive translocation of its cognate intracellular compartment. Muscle denervation also causes a twofold increase in GLUT-1. In normal muscle, all GLUT-1 is present at the cell surface, but in denervated muscle a significant fraction (25.1 +/- 6.1%) of this transporter is found in intracellular vesicles that have the same sedimentation coefficient as GLUT-4-containing vesicles but can be separated from the latter by immunoadsorption. These GLUT-1-containing vesicles respond to insulin and translocate to the cell surface. Thus the formation of insulin-sensitive GLUT-1 containing vesicles in denervated muscle may be a compensatory mechanism for the decreased level of GLUT-4. PMID- 10827004 TI - Role of adrenal renin-angiotensin system in the control of aldosterone secretion in sodium-restricted rats. AB - This study examined the effect of the pharmacological manipulation of adrenal renin-angiotensin system (RAS) on aldosterone secretion from in situ perfused adrenals of rats kept on a normal diet and sodium restricted for 14 days. Neither the angiotensin-converting enzyme inhibitor captopril nor the nonselective angiotensin II receptor antagonist saralasin and the AT(1) receptor-selective antagonist losartan affected basal aldosterone output in normally fed rats. In contrast, they concentration dependently decreased aldosterone secretion in sodium-restricted animals, with maximal effective concentration ranging from 10( 7) to 10(-6) M. Captopril (10(-6) M), saralasin (10(-6) M), and losartan (10(-7) M) counteracted aldosterone response to 10 mM K(+) in sodium-restricted rats but not in normally fed animals. Collectively, these findings provide evidence that adrenal RAS plays a role in the regulation of aldosterone secretion, but only under conditions of prolonged stimulation of zona glomerulosa probably leading to overexpression of adrenal RAS. PMID- 10827005 TI - Peroxynitrite production by TNF-alpha and IL-1beta: implication for suppression of osteoblastic differentiation. AB - To determine the roles of nitric oxide (NO) and its metabolite, peroxynitrite (ONOO(-)), on osteoblastic activation, we investigated the effects of a NO donor [ethanamine, 2, 2'-(hydroxynitrosohydrazono)bis- (dNO)], an O(-2) donor (pyrogallol), and an ONOO(-) scavenger (urate) on alkaline phosphatase (ALPase) activity and osteocalcin gene expression, which are indexes of osteoblastic differentiation. dNO elevated ALPase activity in the osteogenic MC3T3-E1 cell line. The combination of dNO and pyrogallol reduced both ALPase activity and osteocalcin gene expression. Because both indexes were recovered by urate, ONOO( ), unlike NO itself, inhibited the osteoblastic differentiation. Furthermore, treatment with a combination of the proinflammatory cytokines tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) was found to yield ONOO(-) as well as NO and O(-2). The reductions in ALPase activity and osteocalcin gene expression were also restored by urate. We conclude that ONOO(-) produced by TNF-alpha and IL-1beta, but not NO per se, would overcome the stimulatory effect of NO on osteoblastic activity and inhibit osteoblastic differentiation. PMID- 10827006 TI - The oviduct produces erythropoietin in an estrogen- and oxygen-dependent manner. AB - Previously, we showed that erythropoietin (Epo) is produced in the mouse uterus, where Epo is indispensable for estrogen (E(2))-dependent angiogenesis. Expression of uterine Epo mRNA is stimulated by E(2) and hypoxia. The hypoxic induction requires the presence of E(2). In the present study, we examined other female reproductive organs in the mouse with respect to Epo mRNA expression and its stimuli (E(2) and hypoxia)-induced changes. Although Epo mRNA expression was seen in the ovary and oviduct, the E(2)-induced stimulation of Epo mRNA was found only in the oviduct. The E(2)-induced stimulation in the oviduct was transient and rapidly downregulated. Epo mRNA expression in the oviduct was hypoxia inducible, in both the presence and the absence of E(2). E(2)-dependent production of Epo and its mRNA expression were also found by use of cultured oviducts. The E(2) action is probably mediated through the E(2) receptor, and de novo protein synthesis is not required for E(2) induction of Epo mRNA. In the oviduct, the ampulla and isthmus regions produce Epo. PMID- 10827007 TI - Training does not protect against exhaustive exercise-induced lactate transport capacity alterations. AB - The effects of endurance training on lactate transport capacity remain controversial. This study examined whether endurance training 1) alters lactate transport capacity, 2) can protect against exhaustive exercise-induced lactate transport alteration, and 3) can modify heart and oxidative muscle monocarboxylate transporter 1 (MCT1) content. Forty male Wistar rats were divided into control (C), trained (T), exhaustively exercised (E), and trained and exercised (TE) groups. Rats in the T and TE groups ran on a treadmill (1 h/day, 5 days/wk at 25 m/min, 10% incline) for 5 wk; C and E were familiarized with the exercise task for 5 min/day. Before being killed, E and TE rats underwent exhaustive exercise (25 m/min, 10% grade), which lasted 80 and 204 min, respectively (P < 0.05). Although lactate transport measurements (zero-trans) did not differ between groups C and T, both E and TE groups presented an apparent loss of protein saturation properties. In the trained groups, MCT1 content increased in soleus (+28% for T and +26% for TE; P < 0.05) and heart muscle (+36% for T and +33% for TE; P < 0.05). Moreover, despite the metabolic adaptations typically observed after endurance training, we also noted increased lipid peroxidation byproducts after exhaustive exercise. We concluded that 1) endurance training does not alter lactate transport capacity, 2) exhaustive exercise induced lactate transport alteration is not prevented by training despite increased MCT1 content, and 3) exercise-induced oxidative stress may enhance the passive diffusion responsible for the apparent loss of saturation properties, possibly masking lactate transport regulation. PMID- 10827008 TI - Short-term effects of growth hormone on myocardial glucose uptake in healthy humans. AB - Cardiac muscle is characterized by insulin resistance in specific heart diseases such as coronary artery disease and congestive heart failure, but not in generalized disorders like diabetes mellitus and essential hypertension when cardiac manifestations are absent. To examine whether the insulin antagonistic effect of growth hormone (GH) acts upon the heart, we compared insulin-stimulated whole body and myocardial glucose uptake with and without GH administration during a 3.5-h euglycemic-hyperinsulinemic clamp in eight healthy males. Myocardial 2-deoxy-2-[(18)F]fluoro-D-glucose uptake was measured with positron emission tomography. The data were converted to myocardial glucose uptake by tracer kinetic analysis. GH did not change the rate-pressure product. GH decreased whole body insulin-stimulated glucose disposal by 26% (48.0 +/- 12.1 vs. control 62.8 +/- 6.1 micromol. kg(-1). min(-1), P < 0.02). Free fatty acids were suppressed to a similar extent with and without GH during the insulin clamp. Insulin-stimulated myocardial glucose uptake was similar in the presence and in the absence of GH (0.34 +/- 0.05 and 0.31 +/- 0.03 micromol. g(-1). min(-1), P = 0.18). In conclusion, GH does not impair insulin-stimulated myocardial glucose uptake despite a considerable whole body insulin antagonistic effect. Myocardial insulin resistance is not an inherent consequence of whole body insulin resistance. PMID- 10827009 TI - The differential effects of galanin-(1-30) and -(3-30) on anterior pituitary hormone secretion in vivo in humans. AB - Intravenous injection of galanin increases plasma growth hormone (GH) and prolactin (PRL) concentrations. In the rat, the effects of galanin on GH appear to be mediated via the hypothalamic galanin receptor GAL-R(1), at which galanin (3-29) is inactive. In contrast, the effect of galanin on PRL is mediated via the pituitary-specific galanin receptor GAL-R(W), at which galanin-(3-29) is fully active. We investigated the effects of an intravenous infusion of human galanin (hGAL)-(1-30) and -(3-30) on anterior pituitary hormone levels in healthy females. Subjects were infused with saline, hGAL-(1-30) (80 pmol. kg(-1). min( 1)), and hGAL-(3-30) (600 pmol. kg(-1). min(-1)) and with boluses of gonadotropin releasing hormone, thyrotropin-releasing hormone, and growth hormone-releasing hormone (GHRH). Both hGAL-(1-30) and -(3-30) potentiated the rise in GHRH stimulated GH levels [area under the curve (AUC), saline, 2,810 +/- 500 vs. hGAL (1-30), 4,660 +/- 737, P < 0.01; vs. hGAL-(3-30), 6, 870 +/- 1,550 ng. min. ml( 1), P < 0.01]. In contrast to hGAL-(1-30), hGAL-(3-30) had no effect on basal GH levels (AUC, saline, -110 +/- 88 vs. hGAL 1-30, 960 +/- 280, P < 0.002; vs. hGAL (3-30), 110 +/- 54 ng. min. ml(-1), P = not significant). These data suggest that the effects of galanin on basal and stimulated GH release are mediated via different receptor subtypes and that the human equivalent of GAL-R(W) may exist. PMID- 10827010 TI - Abundance and subcellular distribution of MCT1 and MCT4 in heart and fast-twitch skeletal muscles. AB - The expression of two monocarboxylate transporters (MCTs) was examined in muscle and heart. MCT1 and MCT4 proteins are coexpressed in rat skeletal muscles, but only MCT1 is expressed in rat hearts. Among six rat fast-twitch muscles (red and white gastrocnemius, plantaris, extensor digitorum longus, red and white tibialis anterior) there was an inverse relationship between MCT1 and MCT4 (r = -0.94). MCT1 protein was correlated with MCT1 mRNA (r = 0.94). There was no relationship between MCT4 mRNA and MCT4 protein. MCT1 (r = -0.97) and MCT4 (r = 0.88) protein contents were correlated with percent fast-twitch glycolytic fiber. When normalized for their mRNAs, MCT1 but not MCT4 was still correlated with the percent fast-twitch glycolytic fiber composition of rat muscles (r = -0.98). MCT1 and MCT4 were also measured in plasma membranes (PM), triads (TR), T tubules (TT), sarcoplasmic reticulum (SR), and intracellular membranes (IM). There was an intracellular pool of MCT4 but not of MCT1. The MCT1 subcellular distribution was as follows: PM (100%) > TR (31.6%) > SR (15%) = TT (14%) > IM (1.7%). The MCT4 subcellular distribution was considerably different [PM (100%) > TR (66.5%) > TT (36%) = SR (43%) > IM (24%)]. These studies have shown that 1) the mechanisms regulating the expression of MCT1 (transcriptional and posttranscriptional) and MCT4 (posttranscriptional) are different and 2) differences in MCT1 and MCT4 expression among muscles, as well as in their subcellular locations, suggest that they may have different roles in muscle. PMID- 10827011 TI - Exercise attenuates the fasting-induced transcriptional activation of metabolic genes in skeletal muscle. AB - Fasting elicits a progressive increase in lipid metabolism within skeletal muscle. To determine the effects of fasting on the transcriptional regulation of genes important for metabolic control in skeletal muscle composed of different fiber types, nuclei from control and fasted (24 and 72 h) rats were subjected to nuclear run-on analysis using an RT-PCR-based technique. Fasting increased (P < 0.05) transcription rate of the muscle-specific uncoupling protein-3 gene (UCP3) 14.3- to 21.1-fold in white gastrocnemius (WG; fast-twitch glycolytic) and 5.5- to 7.5-fold in red gastrocnemius (RG; fast-twitch oxidative) and plantaris (PL; mixed) muscles. No change occurred in soleus (slow-twitch oxidative) muscle. Fasting also increased transcription rate of the lipoprotein lipase (LPL), muscle carnitine palmitoyltransferase I (CPT I), and long-chain acyl-CoA dehydrogenase (LCAD) genes 1.7- to 3.7-fold in WG, RG, and PL muscles. Transcription rate responses were similar after 24 and 72 h of fasting. Surprisingly, increasing metabolic demand during the initial 8 h of starvation (two 2-h bouts of treadmill running) attenuated the 24-h fasting-induced transcriptional activation of UCP3, LPL, CPT I, and LCAD in RG and PL muscles, suggesting the presence of opposing regulatory mechanisms. These data demonstrate that fasting elicits a fiber type specific coordinate increase in the transcription rate of several genes involved in and/or required for lipid metabolism and indicate that exercise may attenuate the fasting-induced transcriptional activation of specific metabolic genes. PMID- 10827012 TI - Acute response of IGF-I and IGF binding proteins induced by thermal injury. AB - Previous studies demonstrate that thermal injury decreases circulating levels of insulin growth factor I (IGF-I) and alters the plasma concentration of several IGF binding proteins (IGFBP), but the mechanisms for these alterations have not been elucidated. In the current study, a 30% total body surface area full thickness scald burn was produced in anesthetized rats, and animals were studied 24 h later. The plasma concentration of both total and free IGF-I was decreased (38 and 65%, respectively) in burn rats compared with values from time-matched control animals. Thermal injury decreased the IGF-I peptide content in liver approximately 40%, as well as in fast-twitch skeletal muscle (56-69%) and heart (28%). In contrast, IGF-I content in kidney was elevated by 36% in burn rats. Northern blot analysis of liver indicated that burn decreased the expression of small (1.7- and 0.9- to 1.2-kb) IGF-I mRNA transcripts but increased the expression of the 7.5-kb transcript. In contrast, there was a coordinate decrease in all IGF-I mRNA transcripts in muscle and kidney of approximately 30%. For liver, muscle, and kidney, there was no significant difference in the expression of growth hormone receptor mRNA between control and burn rats. Thermal injury increased plasma IGFBP-1 levels, and this change was associated with increased IGFBP-1 mRNA in both liver and kidney. IGFBP-3 levels in plasma were concomitantly decreased by burn injury. This change was associated with a reduction in IGFBP-3 mRNA in liver but an increased expression of IGFBP-3 in kidney and muscle. Thermal injury also decreased the concentration of the acid labile subunit (ALS) in plasma and ALS mRNA expression in liver. Finally, hepatic expression of IGFBP-related peptide-1 was increased twofold in liver but was unchanged in kidney or muscle of burn rats. These results characterize burn induced changes in various components of the IGF system in select tissues and thereby provide potential mechanisms for alterations in the circulating IGF system and for changes in tissue metabolism. PMID- 10827013 TI - Effect of early dietary restriction on insulin action and secretion in the GK rat, a spontaneous model of NIDDM. AB - The availability of the Goto-Kakisaki (GK) rat model of non-insulin-dependent diabetes mellitus prompted us to test the effect of a limited period of undernutrition in previously diabetic young rats on their insulin secretion and insulin action during adult age. Four-week-old female GK rats were either food restricted (35% restriction, 15% protein diet) or protein and energy restricted (35% restriction, 5% protein diet) for 4 wk. Food restriction in the young GK rat lowered weight gain but did not aggravate basal hyperglycemia or glucose intolerance, despite a decrease in basal plasma insulin level. Furthermore, the insulin-mediated glucose uptake by peripheral tissues in the GK rat was clearly improved. We also found that food restriction, when it is coupled to overt protein deficiency in the young GK rat, altered weight gain more severely and slightly decreased basal hyperglycemia but conversely aggravated glucose tolerance. Improvement of basal hyperglycemia was related to repression of basal hepatic glucose hyperproduction, despite profound attenuation of basal plasma insulin level. Deterioration of tolerance to glucose was related to severe blunting of the residual glucose-induced insulin secretion. It is, however, likely that the important enhancement of the insulin-mediated glucose uptake helped to limit the deterioration of glucose tolerance. PMID- 10827014 TI - Immunochemical studies on the putative plasmalemmal receptor for 1, 25(OH)(2)D(3). I. Chick intestine. AB - Antisera were raised against the NH(2)-terminus of the putative basal lateral membrane (BLM) receptor for 1,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3); BLM VDR]. In Western analyses of BLM proteins, antibody (Ab) 099 was monospecific for a 64.5-kDa band. A protein of 64.5 kDa was also labeled by the affinity ligand [(14)C]1, 25(OH)(2)D(3)-bromoacetate; label was diminished in the presence of excess unlabeled secosteroid. The monoclonal antibody against the nuclear VDR (9A7) failed to detect an appropriate band in BLM fractions. Preincubation of isolated intestinal cells with Ab 099, but not 9A7, affected the following two 1,25(OH)(2)D(3)-mediated signal transduction events: augmented intracellular calcium and protein kinase C activity. Subcellular distribution of Ab 099 reactivity by Western analyses and fluorescence microscopy revealed the highest concentrations in BLM followed by the endoplasmic reticulum. Exposure of isolated intestinal cells to 1,25(OH)(2)D(3) for 10 s or vascular perfusion of duodena for 5 min resulted in a time-dependent increase in nuclear localization of the BLM VDR antigen, as judged by electron microscopy, whereas 24, 25-dihydroxyvitamin D(3) failed to increase antigenic labeling in nuclei. Densitometric quantitation of Western blots of subcellular fractions prepared from isolated intestinal cells treated with vehicle or 1,25(OH)(2)D(3) confirmed a hormone-induced increase of putative BLM-VDR in the nucleus. It is concluded that a novel cell surface binding protein for 1,25(OH)(2)D(3) has been identified. PMID- 10827015 TI - Isoproterenol and cAMP regulation of the human brain natriuretic peptide gene involves Src and Rac. AB - Brain natriuretic peptide (BNP) gene expression and chronic activation of the sympathetic nervous system are characteristics of the development of heart failure. We studied the role of the beta-adrenergic signaling pathway in regulation of the human BNP (hBNP) promoter. An hBNP promoter (-1818 to +100) coupled to a luciferase reporter gene was transferred into neonatal cardiac myocytes, and luciferase activity was measured as an index of promoter activity. Isoproterenol (ISO), forskolin, and cAMP stimulated the promoter, and the beta(2) antagonist ICI 118,551 abrogated the effect of ISO. In contrast, the protein kinase A (PKA) inhibitor H-89 failed to block the action of cAMP and ISO. Pertussis toxin (PT), which inactivates Galpha(i), inhibited ISO- and cAMP stimulated hBNP promoter activity. The Src tyrosine kinase inhibitor PP1 and a dominant-negative mutant of the small G protein Rac also abolished the effect of ISO and cAMP. Finally, we studied the involvement of M-CAT-like binding sites in basal and inducible regulation of the hBNP promoter. Mutation of these elements decreased basal and cAMP-induced activity. These data suggest that beta adrenergic regulation of hBNP is PKA independent, involves a Galpha(i)-activated pathway, and targets regulatory elements in the proximal BNP promoter. PMID- 10827016 TI - Trafficking of dietary oleic, linolenic, and stearic acids in fasted or fed lean rats. AB - Increasing evidence supports the notion that there are significant differences in the health effects of diets enriched in saturated, as opposed to monounsaturated or polyunsaturated fat. However, the current understanding of how these types of fat differ in their handling by relevant tissues is incomplete. To examine the effects of fat type and nutritional status on the metabolic fate of dietary fat, we administered (14)C-labeled oleic, linolenic, or stearic acid with a small liquid meal to male Sprague-Dawley rats previously fasted for 15 h (fasted) or previously fed ad libitum (fed). (14)CO(2) production was measured for 8 h after tracer administration. The (14)C content of gastrointestinal tract, serum, liver, skeletal muscle (soleus, lateral, and medial gastrocnemius), and adipose tissue (omental, retroperitoneal, and epididymal) was measured at six time points (2, 4, 8, 24, and 48 h and 10 days) after tracer administration. Plasma levels of glucose, insulin, and triglyceride were also measured. Oxidation of stearic acid was significantly less than that of either linolenic or oleic acid in both the fed and fasted states. This reduction was in part explained by a greater retention of stearic acid within skeletal muscle and liver. Oxidation of oleate and stearate were significantly lower in the fed state than in the fasted state. In the fasted state, liver and skeletal muscle were quantitatively more important than adipose tissue in the uptake of dietary fat tracers during the immediate postprandial period. In contrast, adipose tissue was quantitatively more important than skeletal muscle or liver in the fed state. The movement of carbons derived from dietary fat between tissues is a complex time-dependent process, which varies in response to the type of fat ingested and the metabolic state of the organism. PMID- 10827017 TI - Endotoxin-induced decrease in muscle protein synthesis is associated with changes in eIF2B, eIF4E, and IGF-I. AB - The present study examined potential mechanisms contributing to the inhibition of protein synthesis in skeletal muscle after administration of endotoxin (LPS). Rats implanted with vascular catheters were injected intravenously with a nonlethal dose of Escherichia coli LPS, and samples were collected at 4 and 24 h thereafter; pair-fed control animals were also included. The rate of muscle (gastrocnemius) protein synthesis in vivo was reduced at both time points after LPS administration. LPS did not alter tissue RNA content, but the translational efficiency was consistently reduced at both time points. To identify mechanisms responsible for regulating translation, we examined several eukaryotic initiation factors (eIFs). The content of eIF2alpha or the amount of eIF2alpha in the phosphorylated form did not change in response to LPS. eIF2B activity was decreased in muscle 4 h post-LPS but activity returned to control values by 24 h. A decrease in the relative amount of eIF2Balpha protein was not responsible for the LPS-induced reduction in eIF2B activity. LPS also markedly altered the distribution of eIF4E in muscle. Compared with control values, LPS-treated rats demonstrated 1) a transient increase in binding of the translation repressor 4E binding protein-1 (4E-BP1) with eIF4E, 2) a transient decrease in the phosphorylated gamma-form of 4E-BP1, and 3) a sustained decrease in the amount of eIF4G associated with eIF4E. LPS also decreased insulin-like growth factor (IGF) I protein and mRNA expression in muscle at both times. A significant linear relationship existed between muscle IGF-I and the rate of protein synthesis or the amount of eIF4E bound to eIF4G. In summary, these data suggest that LPS impairs muscle protein synthesis, at least in part, by decreasing translational efficiency, resulting from an impairment in translation initiation associated with alterations in both eIF2B activity and eIF4E availability. PMID- 10827018 TI - Whole body and abdominal lipolytic sensitivity to epinephrine is suppressed in upper body obese women. AB - We measured whole body and regional lipolytic and adipose tissue blood flow (ATBF) sensitivity to epinephrine in 8 lean [body mass index (BMI): 21 +/- 1 kg/m(2)] and 10 upper body obese (UBO) women (BMI: 38 +/- 1 kg/m(2); waist circumference >100 cm). All subjects underwent a four-stage epinephrine infusion (0.00125, 0.005, 0.0125, and 0.025 microgram. kg fat-free mass(-1). min(-1)) plus pancreatic hormonal clamp. Whole body free fatty acid (FFA) and glycerol rates of appearance (R(a)) in plasma were determined by stable isotope tracer methodology. Abdominal and femoral subcutaneous adipose tissue lipolytic activity was determined by microdialysis and (133)Xe clearance methods. Basal whole body FFA R(a) and glycerol R(a) were both greater (P < 0.05) in obese (449 +/- 31 and 220 +/- 12 micromol/min, respectively) compared with lean subjects (323 +/- 44 and 167 +/- 21 micromol/min, respectively). Epinephrine infusion significantly increased FFA R(a) and glycerol R(a) in lean (71 +/- 21 and 122 +/- 52%, respectively; P < 0.05) but not obese subjects (7 +/- 6 and 39 +/- 10%, respectively; P = not significant). In addition, lipolytic and ATBF sensitivity to epinephrine was blunted in abdominal but not femoral subcutaneous adipose tissue of obese compared with lean subjects. We conclude that whole body lipolytic sensitivity to epinephrine is blunted in women with UBO because of decreased sensitivity in upper body but not lower body subcutaneous adipose tissue. PMID- 10827019 TI - Differences in skeletal and muscle mass with aging in black and white women. AB - Previous cross-sectional studies using delayed gamma neutron activation analysis and whole body counting suggested that the relationship of total body calcium (TBCa) to total body potassium (TBK) (muscle mass, body cell mass) remained constant with age. This led to the hypothesis that the muscle mass and skeletal mass compartments are integrated in their response to aging. It had also been hypothesized that loss of skeletal and muscle mass was similar between races. In the current study, delayed gamma neutron activation analysis and whole body counting were performed on 90 black and 143 white women 20-69 yr of age. Black women had higher TBCa and TBK values than white women, even when the data were adjusted for age, height, and weight. TBCa was correlated with height and TBK with weight. The estimated decline of skeletal mass (TBCa) from 20 to 70 yr was 18% in black women and 19% in white women. However, the lifetime decline of TBK was only 8% for black women, compared with 22% for white women. Black women may lose TBK more slowly than TBCa with aging, compared with white women. In particular, correlation of TBCa and age was similar for blacks and whites (r = 0.44 and r = -0.54, respectively). However, for TBK these correlations were r = 0.14 and r = -0.42. These data confirm a higher musculoskeletal mass in black women and suggest that the loss of muscle mass with age may be lower in black than in white women. These ethnic differences do not support the hypothesis of an integrated musculoskeletal system, so that these two components should be considered separately. A prospective study is needed to confirm these findings. PMID- 10827020 TI - Partial saturation and regional variation in the blood-to-brain transport of leptin in normal weight mice. AB - Impaired blood-brain barrier transport of leptin into the arcuate nucleus has been suggested to underlie obesity in humans and outbred aging mice. Here, we used a brain perfusion method in mice to measure transport rates and kinetic parameters for leptin at vascular concentrations between 0.15 and 130 ng/ml. Transport into whole brain was partially saturated at all concentrations, not only those seen in obesity. Leptin entered all regions of the brain, not only the hypothalamus, with entry and saturation rates differing among the brain regions. The value of the Michaelis-Menten constant of the transporter approximates normal serum levels and the maximum velocity value varies significantly among brain regions. These results suggest an important role for low serum levels signaling starvation status to the brain and show that the levels of leptin seen in obesity greatly saturate the transporter. Differences in regional uptake and saturation provide a mechanism by which leptin can control events mediated at the arcuate nucleus and other regions of the central nervous system with different regional thresholds for optimal function. PMID- 10827021 TI - Control of ovine hepatic growth hormone receptor and insulin-like growth factor I by thyroid hormones in utero. AB - By use of RNase protection assays, hepatic growth hormone receptor (GHR) and insulin-like growth factor I (IGF-I) mRNA abundances were measured in sheep fetuses after experimental manipulation of fetal plasma thyroid hormone concentrations by fetal thyroidectomy (TX) and exogenous infusion of triiodothyronine (T(3)) and cortisol. TX abolished the normal prepartum rise in hepatic GHR abundance but had little effect on hepatic GHR gene expression at 127 130 days (term 145 +/- 2 days). By contrast, it upregulated basal IGF-I expression in immature fetal liver by increasing both Class 1 and Class 2 transcript abundance but had no further effects on IGF-I gene mRNA levels at 142 145 days. Raising plasma T(3) to prepartum values by exogenous infusion of either T(3) or cortisol into immature intact fetuses prematurely raised hepatic GHR and IGF-I mRNA abundances to values similar to those seen in intact fetuses at 142 145 days. In TX fetuses, cortisol infusion increased hepatic GHR mRNA but not total IGF-I mRNA abundance at 127-130 days. These findings show that thyroid hormones have an important role in the regulation of hepatic GHR and IGF-I gene expression in fetal sheep during late gestation and suggest that T(3) mediates the maturational effects of cortisol on the hepatic somatotropic axis close to term. PMID- 10827022 TI - Changing society: changing role of doctors. The stresses must not be allowed to get too great. PMID- 10827023 TI - Fluctuations in lower urinary tract symptoms in women. Reassurance and watchful waiting can prevent overtreatment. PMID- 10827024 TI - Magnetic resonance imaging of the pelvis. Better techniques can help determine management and predict outcome. PMID- 10827025 TI - Distinguishing mental illness in primary care. We need to separate proper syndromes from generalised distress. PMID- 10827026 TI - Dereliction of duty in an ageist society. The government's silence over Royal Commission report on long term care is ominous. PMID- 10827028 TI - Juniors to wait 9 years for EU rules to bite PMID- 10827027 TI - Paediatric surgery should be performed by specialist surgeons. PMID- 10827029 TI - Appeal court refuses mother's request for hysterectomy. PMID- 10827030 TI - Digital radio relays text to remote doctors and nurses in Africa. PMID- 10827031 TI - In brief PMID- 10827032 TI - Israeli infertility experts investigated for "selling" ova. PMID- 10827033 TI - GMC to consult on next stage of revalidation. PMID- 10827035 TI - Proceedings start against former chemical warfare chief. PMID- 10827034 TI - AIDS activists arrested in India. PMID- 10827036 TI - Milburn attacks doctors' private pay PMID- 10827037 TI - Tobacco firms fight ban on advertisements PMID- 10827038 TI - Campaigners accuse tobacco firm of dubious ploy. PMID- 10827039 TI - FDA approves device for female sexual dysfunction. PMID- 10827041 TI - Deaths from breast cancer fall dramatically in UK and US PMID- 10827040 TI - Thrombolytic agents no benefit to those over 75 PMID- 10827042 TI - Incidence and remission rates of lower urinary tract symptoms at one year in women aged 40-60: longitudinal study. AB - OBJECTIVES: To determine the incidence and rates of remission of lower urinary tract symptoms at one year in women aged 40-60, and to assess factors associated with remission. DESIGN: Ongoing longitudinal cohort study. SETTING: One rural and one urban county in Denmark. PARTICIPANTS: 4000 women recruited on a random basis, 2860 of whom were followed up at one year. MEASUREMENTS: Incidence and rates of remission of lower urinary tract symptoms. RESULTS: Prevalence, incidence, and rates of remission of lower urinary tract symptoms in 2284 women were respectively 28.5% (95% confidence interval 26.7% to 30.4%), 10.0% (8.5% to 11.4%), and 27.8% (25.6% to 30.0%). Overall, symptoms were not significantly associated with events performed or initiated in the study period: medical consultation (1.6, 0.8 to 2.8), pelvic floor physiotherapy (0.9, 0.5 to 1.8), treatment with antibiotics on suspicion of a lower urinary tract infection (1.3, 0.8 to 2.2), or other treatment (1.7, 0.7 to 4. 1). Many of the individual symptoms were, however, associated with seeking professional help. CONCLUSIONS: Lower urinary tract symptoms constitute dynamic conditions, with women experiencing more or fewer symptoms, and eventually a cessation of symptoms. The distinction between permanent and fluctuating cases may have important clinical and scientific implications. PMID- 10827043 TI - Association between psychosocial work characteristics and health functioning in American women: prospective study. AB - OBJECTIVE: To examine prospectively the relation between psychosocial work characteristics and changes in health related quality of life over four years in a cohort of working women in the United States. DESIGN: Longitudinal cohort study. SETTING: United States. PARTICIPANTS: 21 290 female registered nurses who completed the Karasek's job content questionnaire and a modified version of the short form 36 questionnaire (SF-36) as used for a survey of health status by the medical outcomes study. MAIN OUTCOME MEASURES: Seven dimensions of health status: physical functioning, role limitations due to physical health problems, bodily pain, vitality, social functioning, role limitations due to emotional problems, and mental health. RESULTS: Examined separately low job control, high job demands, and low work related social support were associated with poor health status at baseline as well as greater functional declines over the four year follow up period. Examined in combination, women with low job control, high job demands, and low work related social support ("iso-strain" jobs) had the greatest functional declines. These associations could not be explained by age, body mass index, comorbid disease status, alcohol consumption, smoking status, education level, exercise level, employment status, marital status, or presence of a confidant. CONCLUSIONS: Adverse psychosocial work conditions are important predictors of poor functional status and its decline over time. PMID- 10827044 TI - Relation between a career and family life for English hospital consultants: qualitative, semistructured interview study. AB - OBJECTIVE: To explore the relation between work and family life among hospital consultants and their attitude towards the choices and constraints that influence this relation. DESIGN: Qualitative study of consultants' experiences and views based on tape recorded semistructured interviews. SETTING: Former South Thames health region in southeast England. PARTICIPANTS: 202 male and female NHS hospital consultants aged between 40 to 50 years representing all hospital medical specialties. RESULTS: Three types of relation between work and family life (career dominant, segregated, and accommodating) were identified among hospital consultants. Most consultants had a segregated relation, although female consultants were more likely than male consultants to have a career dominant or an accommodating relation. Many male consultants and some female consultants expressed considerable dissatisfaction with the balance between their career and family life. A factor influencing this dissatisfaction was the perceived lack of choice to spend time on their personal or family life, because of the working practices and attitudes within hospital culture, if they wanted a successful career. CONCLUSIONS: Consultants are currently fitting in with the profession rather than the profession adapting to enable doctors to have fulfilling professional and personal lives. Current government policies to increase the medical workforce and promote family friendly policies in the NHS ought to take account of the need for a fundamental change in hospital culture to enable doctors to be more involved in their personal or family life without detriment to their career progress. PMID- 10827046 TI - Email submissions from outside the united kingdom PMID- 10827045 TI - Hospital admissions for acute anaphylaxis: time trend study. PMID- 10827047 TI - Primary and secondary care management of women with early breast cancer from affluent and deprived areas: retrospective review of hospital and general practice records. AB - OBJECTIVES: To investigate whether poorer survival of breast cancer among deprived women compared with affluent women is related to their NHS care. DESIGN: Retrospective review of hospital and general practice case records. SETTING: Greater Glasgow Health Board area. SUBJECTS: Women diagnosed with breast cancer in 1992-3 who lived in the most affluent (deprivation categories 1 and 2) and the most deprived areas (deprivation categories 6 and 7) of Glasgow (Carstairs and Morris deprivation index). MAIN OUTCOME MEASURES: Breast cancer treatment, time from general practice consultation to clinic visit and surgery, and details of hospital admissions and follow up in primary and secondary care. RESULTS: The access to care and surgical and oncological treatment of women from affluent and deprived areas were similar. Admissions to hospital for problems not related to breast cancer were more common in those living in deprived areas (number admitted once or more: 51 (24%) v 13 (10%), P=0.001). Consultation patterns in general practice by the second year after diagnosis showed women in deprived areas consulting more frequently than women in affluent areas (median (interquartile range) number of consultations (5 (2-10) v 7 (4-13), P=0.01). CONCLUSION: Women living in affluent areas did not receive better NHS care for breast cancer than women in deprived areas. However, women from deprived areas seem to have greater comorbidity, and poorer outcomes from breast cancer among these women is probably due to factors which result in deprived communities having poorer health outcomes rather than to management of their breast cancer. PMID- 10827048 TI - Listening to the patient PMID- 10827049 TI - Thresholds for taking antihypertensive drugs in different professional and lay groups: questionnaire survey. PMID- 10827050 TI - Aggression and violent behaviour in general practice: population based survey in the north of England. PMID- 10827051 TI - Books of life, death-and what comes in between PMID- 10827053 TI - Check, check, and check again PMID- 10827052 TI - Extracts from the "clinical evidence". Endometriosis. AB - DEFINITION: Endometriosis is characterised by ectopic endometrial tissue, which can cause dysmenorrhoea, dyspareunia, non-cyclical pelvic pain, and subfertility. Diagnosis is made by laparoscopy. Most endometrial deposits are found in the pelvis (ovaries, peritoneum, uterosacral ligaments, pouch of Douglas, and rectovaginal septum). Extrapelvic deposits, including those in the umbilicus and diaphragm, are rare. Endometriomas are cysts of endometriosis within the ovary. INCIDENCE/PREVALENCE: In asymptomatic women, the prevalence ranges from 2% to 22%, depending on the diagnostic criteria used and the populations studied. In women with dysmenorrhoea, the incidence of endometriosis ranges from 40% to 60%, and in women with subfertility it ranges from 20% to 30%. The severity of symptoms and the probability of diagnosis increase with age. Incidence peaks at about age 40. Symptoms and laparoscopic appearance do not always correlate. AETIOLOGY: The cause is unknown. Risk factors include early menarche and late menopause. Embryonic cells may give rise to deposits in the umbilicus, while retrograde menstruation may deposit endometrial cells in the diaphragm. Oral contraceptives reduce the risk of endometriosis, and this protective effect persists for up to a year after their discontinuation. PROGNOSIS: We found one small randomised controlled trial (RCT) in which repeat laparoscopy was performed in the women treated with placebo. Over 12 months, endometrial deposits resolved spontaneously in a quarter, deteriorated in nearly half, and were unchanged in the remainder. AIMS: To relieve pain (dysmenorrhoea, dyspareunia, and other pelvic pain) and to improve fertility, with minimal adverse effects. OUTCOMES: American Fertility Society scores for size and number of deposits; recurrence rates; time between stopping treatment and recurrence; rate of adverse effects of treatment. In women with pain: relief of pain, assessed by visual analogue scale and subjective improvement. In women with subfertility: cumulative pregnancy rate, live birth rate. In women undergoing surgery: ease of surgical intervention (rated as easy, average, difficult, or very difficult). PMID- 10827055 TI - Understanding today's joads PMID- 10827054 TI - ABC of arterial and venous disease. Swollen lower limb-1: general assessment and deep vein thrombosis. PMID- 10827056 TI - The secret life of the NHS. PMID- 10827057 TI - The private finance initiative: spinning out the defence. PMID- 10827058 TI - Modernizing the NHS. Performance and productivity. PMID- 10827060 TI - Not the right welcome PMID- 10827059 TI - Measuring performance in the NHS: what really matters? PMID- 10827061 TI - Statistics notes. The odds ratio. PMID- 10827062 TI - Business of the philosopher PMID- 10827063 TI - Obituaries PMID- 10827064 TI - PFI poses threat to health care PMID- 10827065 TI - New labour, new language? PMID- 10827066 TI - Caring for women with circumcision: A technical manual for health care providers PMID- 10827068 TI - Vilified for tackling tobacco PMID- 10827067 TI - The river: A journey back to the source of HIV and AIDS PMID- 10827069 TI - Urinary tract symptoms PMID- 10827070 TI - Has humanity disappeared from the NHS? PMID- 10827071 TI - Be very afraid PMID- 10827072 TI - New labour, new stalinism PMID- 10827073 TI - Lower urinary tract symptoms follow a variable course PMID- 10827075 TI - Consultants feel imbalance between work and family life PMID- 10827076 TI - Admissions for acute anaphylaxis have increased in england PMID- 10827074 TI - Low job control leads to decline in physical function and mental health PMID- 10827077 TI - Women from deprived areas do not receive worse treatment for breast cancer PMID- 10827078 TI - GPs' susceptibility to abuse varies with deprivation PMID- 10827079 TI - HEDJ, an Hsp40 co-chaperone localized to the endoplasmic reticulum of human cells. AB - Hsp40 co-chaperones, characterized by the presence of a highly conserved J domain, are involved in nearly all aspects of protein synthesis, folding, and secretion. Within the lumen of the endoplasmic reticulum, these chaperones are also involved in reverse translocation and degradation of misfolded proteins. We describe here the cloning and characterization of a novel Hsp40 chaperone, which we named HEDJ. Epitope-tagged HEDJ was demonstrated by confocal microscopy to be localized to the endoplasmic reticulum. Protease susceptibility, glycosidase treatment, and detergent solubility assays demonstrated that the molecule was luminally oriented and membrane-associated. In vitro experiments demonstrated that the J domain interacted with the endoplasmic reticulum-associated Hsp70, Bip, in an ATP-dependent manner and was capable of stimulating its ATPase activity. HEDJ mRNA expression was detected in all human tissues examined. Highly homologous sequences were found in mouse, Drosophila, and Caenorhabditis elegans data bases. These results suggest potential roles for HEDJ in protein import, folding, or translocation within the endoplasmic reticulum. PMID- 10827080 TI - Mammalian peptidoglycan recognition protein binds peptidoglycan with high affinity, is expressed in neutrophils, and inhibits bacterial growth. AB - Peptidoglycan recognition protein (PGRP) is conserved from insects to mammals. In insects, PGRP recognizes bacterial cell wall peptidoglycan (PGN) and activates prophenoloxidase cascade, a part of the insect antimicrobial defense system. Because mammals do not have the prophenoloxidase cascade, its function in mammals is unknown. However, it was suggested that an identical protein (Tag7) was a tumor necrosis factor-like cytokine. Therefore, the aim of this study was to identify the function of PGRP in mammals. Mouse PGRP bound to PGN with fast kinetics and nanomolar affinity (K(d) = 13 nm). The binding was specific for polymeric PGN or Gram-positive bacteria with unmodified PGN, and PGRP did not bind to other cell wall components or Gram-negative bacteria. PGRP mRNA and protein were expressed in neutrophils and bone marrow cells, but not in spleen cells, mononuclear cells, T or B lymphocytes, NK cells, thymocytes, monocytes, and macrophages. PGRP was not a PGN-lytic or a bacteriolytic enzyme, but it inhibited the growth of Gram-positive but not Gram-negative bacteria. PGRP inhibited phagocytosis of Gram-positive bacteria by macrophages, induction of oxidative burst by Gram-positive bacteria in neutrophils, and induction of cytokine production by PGN in macrophages. PGRP had no tumor necrosis factor-like cytotoxicity for mammalian cells, and it was not chemotactic on its own or in combination with PGN. Therefore, mammalian PGRP binds to PGN and Gram-positive bacteria with nanomolar affinity, is expressed in neutrophils, and inhibits growth of bacteria. PMID- 10827081 TI - NIPP1-mediated interaction of protein phosphatase-1 with CDC5L, a regulator of pre-mRNA splicing and mitotic entry. AB - NIPP1 is a regulatory subunit of a species of protein phosphatase-1 (PP1) that co localizes with splicing factors in nuclear speckles. We report that the N terminal third of NIPP1 largely consists of a Forkhead-associated (FHA) protein interaction domain, a known phosphopeptide interaction module. A yeast two-hybrid screening revealed an interaction between this domain and a human homolog (CDC5L) of the fission yeast protein cdc5, which is required for G(2)/M progression and pre-mRNA splicing. CDC5L and NIPP1 co-localized in nuclear speckles in COS-1 cells. Furthermore, an interaction between CDC5L, NIPP1, and PP1 in rat liver nuclear extracts could be demonstrated by co-immunoprecipitation and/or co purification experiments. The binding of the FHA domain of NIPP1 to CDC5L was dependent on the phosphorylation of CDC5L, e.g. by cyclin E-Cdk2. When expressed in COS-1 or HeLa cells, the FHA domain of NIPP1 did not affect the number of cells in the G(2)/M transition. However, the FHA domain blocked beta-globin pre mRNA splicing in nuclear extracts. A mutation in the FHA domain that abolished its interaction with CDC5L also canceled its anti-splicing effects. We suggest that NIPP1 either targets CDC5L or an associated protein for dephosphorylation by PP1 or serves as an anchor for both PP1 and CDC5L. PMID- 10827082 TI - Amino acid residues conferring ligand binding properties of prostaglandin I and prostaglandin D receptors. Identification by site-directed mutagenesis. AB - Using chimeras of the mouse prostaglandin (PG) I receptor (mIP) and the mouse PGD receptor (mDP), we previously revealed that the cyclopentane ring recognition by these receptors is specified by a region from the first to third transmembrane domain of each receptor; recognition by this region of mIP is broad, accommodating the D, E, and I types of cyclopentane rings, whereas that of mDP binds the D type of PGs alone (Kobayashi, T., Kiriyama, M., Hirata, T., Hirata, M., Ushikubi, F., and Narumiya, S. (1997) J. Biol. Chem. 272, 15154-15160). In the present study, we performed a more detailed chimera analysis, and narrowed the domain for the ring recognition to a region from the first transmembrane domain to the first extracellular loop. One chimera with the replacement of the second transmembrane domain and the first extracellular loop of mDP with that of mIP bound only iloprost. The amino acid substitutions in this chimera suggest that Ser(50) in the first transmembrane domain of mIP confers the broad ligand recognition of mIP and that Lys(75) and Leu(83) in the second transmembrane domain of mDP confer the high affinity to PGD(2) and the strict specificity of ligand binding of mDP, respectively. PMID- 10827083 TI - A novel double-headed proteinaceous inhibitor for metalloproteinase and serine proteinase. AB - A novel proteinaceous inhibitor for the metalloproteinase of Streptomyces caespitosus has been isolated from the culture supernatant of Streptomyces sp. I 355. It was named ScNPI (Streptomyces caespitosus neutral proteinase inhibitor). ScNPI exhibited strong inhibitory activity toward ScNP with a K(i) value of 1.6 nm. In addition, ScNPI was capable of inhibiting subtilisin BPN' (K(i) = 1.4 nm) (EC ). The scnpi gene consists of two regions, a signal peptide (28 amino acid residues) and a mature region (113 amino acid residues, M(r) = 11,857). The deduced amino acid sequence of scnpi showed high similarity to those of Streptomyces subtilisin inhibitor (SSI) and its homologues. The reactive site of ScNPI for inhibition of subtilisin BPN' was identified to be Met(71)-Tyr(72) bond by specific cleavage. To identify the reactive site for ScNP, Tyr(33) and Tyr(72), which are not conserved among other SSI family inhibitors but are preferable amino acid residues for ScNP, were replaced separately by Ala. The Y33A mutant retained inhibitory activity toward subtilisin BPN' but did not show any inhibitory activity toward ScNP. Moreover, a dimer of ternary complexes among ScNPI, ScNP, and subtilisin BPN' was formed to give the 2:2:2 stoichiometry. These results strongly indicate that ScNPI is a double-headed inhibitor that has individual reactive sites for ScNP and subtilisin BPN'. PMID- 10827084 TI - Conserved serine and histidine residues are critical for activity of the ER-type signal peptidase SipW of Bacillus subtilis. AB - Type I signal peptidases (SPases) are required for the removal of signal peptides from translocated proteins and, subsequently, release of the mature protein from the trans side of the membrane. Interestingly, prokaryotic (P-type) and endoplasmic reticular (ER-type) SPases are functionally equivalent, but structurally quite different, forming two distinct SPase families that share only few conserved residues. P-type SPases were, so far, exclusively identified in eubacteria and organelles, whereas ER-type SPases were found in the three kingdoms of life. Strikingly, the presence of ER-type SPases appears to be limited to sporulating Gram-positive eubacteria. The present studies were aimed at the identification of potential active site residues of the ER-type SPase SipW of Bacillus subtilis, which is required for processing of the spore-associated protein TasA. Conserved serine, histidine, and aspartic acid residues are critical for SipW activity, suggesting that the ER-type SPases employ a Ser-His Asp catalytic triad or, alternatively, a Ser-His catalytic dyad. In contrast, the P-type SPases employ a Ser-Lys catalytic dyad (Paetzel, M., Dalbey, R. E., and Strynadka, N. C. J. (1998) Nature 396, 186-190). Notably, catalytic activity of SipW was not only essential for pre-TasA processing, but also for the incorporation of mature TasA into spores. PMID- 10827086 TI - Subtype-specific translocation of diacylglycerol kinase alpha and gamma and its correlation with protein kinase C. AB - We examined the translocation of diacylglycerol kinase (DGK) alpha and gamma fused with green fluorescent protein in living Chinese hamster ovary K1 cells (CHO-K1) and investigated temporal and spatial correlations between DGK and protein kinase C (PKC) when both kinases are overexpressed. DGKalpha and gamma were present throughout the cytoplasm of CHO-K1 cells. Tetradecanoylphorbol 13 acetate (TPA) induced irreversible translocation of DGKgamma, but not DGKalpha, from the cytoplasm to the plasma membrane. The (TPA)-induced translocation of DGKgamma was inhibited by the mutation of C1A but not C1B domain of DGKgamma and was not inhibited by staurosporine. Arachidonic acid induced reversible translocation of DGKgamma from the cytoplasm to the plasma membrane, whereas DGKalpha showed irreversible translocation to the plasma membrane and the Golgi network. Purinergic stimulation induced reversible translocation of both DGKgamma and alpha to the plasma membrane. The timing of the ATP-induced translocation of DGKgamma roughly coincided with that of PKCgamma re-translocation from the membrane to the cytoplasm. Furthermore, re-translocation of PKCgamma was obviously hastened by co-expression with DGKgamma and was blocked by an inhibitor of DGK (R59022). These results indicate that DGK shows subtype-specific translocation depending on extracellular signals and suggest that PKC and DGK are orchestrated temporally and spatially in the signal transduction. PMID- 10827085 TI - Kinetic and spectroscopic evidence for three actomyosin:ADP states in smooth muscle. AB - Smooth muscle myosin II undergoes an additional movement of the regulatory domain with ADP release that is not seen with fast skeletal muscle myosin II. In this study, we have examined the interactions of smooth muscle myosin subfragment 1 with ADP to see if this additional movement corresponds to an identifiable state change. These studies indicate that for this myosin:ADP, both the catalytic site and the actin-binding site can each assume one of two conformations. Relatively loose coupling between these two binding sites leads to three discrete actin associated ADP states. Following an initial, weakly bound state, binding of myosin:ADP to actin shifts the equilibrium toward a mixture of two states that each bind actin strongly but differ in the conformation of their catalytic sites. By contrast, fast myosins, including Dictyostelium myosin II, have reciprocal coupling between the actin- and ADP-binding sites, so that either actin or nucleotide, but not both, can be tightly bound. This uncoupling, which generates a second strongly bound actomyosin ADP state in smooth muscle, would prolong the fraction of the ATPase cycle time that this actomyosin spends in a force generating conformation and may be central to explaining the physiologic differences between this and other myosins. PMID- 10827087 TI - Sensitization to death receptor cytotoxicity by inhibition of fas-associated death domain protein (FADD)/caspase signaling. Requirement of cell cycle progression. AB - Upon binding of their ligands, death receptors belonging to the tumor necrosis factor (TNF) receptor family initiate a signaling pathway leading to the activation of caspases and ultimately apoptosis. TNF, however, in parallel elicits survival signals, protecting many cell types from cell death that can only be induced by combined treatment with TNF and inhibitors of protein synthesis. Here, we report that in NIH3T3 cells, apoptosis in response TNF and cycloheximide is not inhibited by the broad spectrum caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone (zVAD. fmk). Moreover, treatment with zVAD.fmk sensitizes the cells to the cytotoxic action of TNF. Sensitization was also achieved by overexpression of a dominant-negative mutant of Fas-associated death domain protein and, to a lesser extent, by specific inhibition of caspase-8. A similar, but weaker sensitization of zVAD.fmk to treatment with the TNF-related apoptosis-inducing ligand (TRAIL) or anti-CD95 antibody was demonstrated. The unexpected cell death in response to TNF and caspase inhibition occurs despite the activation of nuclear factor kappaB and c Jun N-terminal kinases. The mode of cell death shows several signs of apoptosis including DNA fragmentation, although activation of caspase-3 was excluded. TNF/zVAD.fmk-induced cell death is preceded by an accumulation of cells in the G(2)/M phase of the cell cycle, indicating an important role of cell cycle progression. This hypothesis is further strengthened by the observation that arresting the cells in the G(1) phase of the cell cycle inhibited TNF/zVAD.fmk induced cell death, whereas blocking them in the G(2)/M phase augmented it. PMID- 10827088 TI - Substitutions in a homologous region of extracellular loop 2 of CXCR4 and CCR5 alter coreceptor activities for HIV-1 membrane fusion and virus entry. AB - CXCR4 and CCR5 are the principal coreceptors for human immunodeficiency virus type-1 (HIV-1) infection. Previously, mutagenesis of CXCR4 identified single amino acid changes that either impaired CXCR4's coreceptor activity for CXCR4 dependent (X4) isolate envelope glycoproteins (Env) or expanded its activity, allowing it to serve as a functional coreceptor for CCR5-dependent (R5) isolates. The most potent of these point mutations was an alanine substitution for the aspartic acid residue at position 187 in extracellular loop 2 (ecl-2), and here we show that this mutation also permits a variety of primary R5 isolate Envs, including those of other subtypes (clades), to employ it as a coreceptor. We also examined the corresponding region of CCR5 and demonstrate that the substitution of the serine residue in the homologous ecl-2 position with aspartic acid impairs CCR5 coreceptor activity for isolates across several clades. These results highlight a homologous and critical element in ecl-2, of both the CXCR4 and CCR5 molecules, for their respective coreceptor activities. Charge elimination expands CXCR4 coreceptor activity, while a similar charge introduction can destroy the coreceptor function of CCR5. These findings provide further evidence that there are conserved elements in both CXCR4 and CCR5 involved in coreceptor function. PMID- 10827089 TI - Gram-negative bacteria-binding protein, a pattern recognition receptor for lipopolysaccharide and beta-1,3-glucan that mediates the signaling for the induction of innate immune genes in Drosophila melanogaster cells. AB - Pattern recognition receptors, non-clonal immune proteins recognizing common microbial components, are critical for non-self recognition and the subsequent induction of Rel/NF-kappaB-controlled innate immune genes. However, the molecular identities of such receptors are still obscure. Here, we present data showing that Drosophila possesses at least three cDNAs encoding members of the Gram negative bacteria-binding protein (DGNBP) family, one of which, DGNBP-1, has been characterized. Western blot, flow cytometric, and confocal laser microscopic analyses demonstrate that DGNBP-1 exists in both a soluble and a glycosylphosphatidylinositol-anchored membrane form in culture medium supernatant and on Drosophila immunocompetent cells, respectively. DGNBP-1 has a high affinity to microbial immune elicitors such as lipopolysaccharide (LPS) and beta 1,3-glucan whereas no binding affinity is detected with peptidoglycan, beta-1,4 glucan, or chitin. Importantly, the overexpression of DGNBP-1 in Drosophila immunocompetent cells enhances LPS- and beta-1,3-glucan-induced innate immune gene (NF-kappaB-dependent antimicrobial peptide gene) expression, which can be specifically blocked by pretreatment with anti-DGNBP-1 antibody. These results suggest that DGNBP-1 functions as a pattern recognition receptor for LPS from Gram-negative bacteria and beta-1, 3-glucan from fungi and plays an important role in non-self recognition and the subsequent immune signal transmission for the induction of antimicrobial peptide genes in the Drosophila innate immune system. PMID- 10827090 TI - The alpha, but not the beta, isoform of the human thromboxane A2 receptor is a target for prostacyclin-mediated desensitization. AB - In this study, we examined the effects the prostacyclin receptor (IP) agonist cicaprost exhibited on U46619-mediated thromboxane A(2) receptor (TP) signaling in platelets and compared it to that which occurs in human embryonic kidney (HEK) 293 cells stably overexpressing the individual TPalpha or TPbeta isoforms. Consistent with previous studies, cicaprost abrogated U46619-mediated platelet aggregation and mobilization of intracellular calcium ([Ca(2+)](i)). In HEK 293 cells, signaling by TPalpha, but not TPbeta, was subject to IP-mediated desensitization in a protein kinase A-dependent, protein kinase C-independent manner. Desensitization of TPalpha signaling was independent of the nature of the IP agonist used, the level of IP expression, or the subtype of G(q) protein. Signaling by TP(Delta)(328), a truncated variant of TP devoid of the divergent residues of the TPs, or by TPalpha(S329A), a site-directed mutant of TPalpha, were insensitive to IP agonist activation. Whole cell phosphorylations established that TPalpha, but not TPbeta or TPalpha(S329A), is subject to IP mediated phosphorylation and that TPalpha phosphorylation is inhibited by H-89. Thus, we conclude that TPalpha, but not TPbeta, is subject to cross desensitization by IP mediated through direct protein kinase A phosphorylation at Ser(329) and propose that TPalpha may be the isoform physiologically relevant to TP:IP-mediated vascular hemostasis. PMID- 10827091 TI - Spectroscopic investigation of selective cluster conversion of archaeal zinc containing ferredoxin from Sulfolobus sp. strain 7. AB - Archaeal zinc-containing ferredoxin from Sulfolobus sp. strain 7 contains one [3Fe-4S] cluster (cluster I), one [4Fe-4S] cluster (cluster II), and one isolated zinc center. Oxidative degradation of this ferredoxin led to the formation of a stable intermediate with 1 zinc and approximately 6 iron atoms. The metal centers of this intermediate were analyzed by electron paramagnetic resonance (EPR), low temperature resonance Raman, x-ray absorption, and (1)H NMR spectroscopies. The spectroscopic data suggest that (i) cluster II was selectively converted to a cubane [3Fe-4S](1+) cluster in the intermediate, without forming a stable radical species, and that (ii) the local metric environments of cluster I and the isolated zinc site did not change significantly in the intermediate. It is concluded that the initial step of oxidative degradation of the archaeal zinc containing ferredoxin is selective conversion of cluster II, generating a novel intermediate containing two [3Fe-4S] clusters and an isolated zinc center. At this stage, significant structural rearrangement of the protein does not occur. We propose a new scheme for oxidative degradation of dicluster ferredoxins in which each cluster converts in a stepwise manner, prior to apoprotein formation, and discuss its structural and evolutionary implications. PMID- 10827092 TI - Inhibitory and stimulatory effects of lactacystin on expression of nitric oxide synthase type 2 in brain glial cells. The role of Ikappa B-beta. AB - Expression of inflammatory nitric oxide synthase (NOS2) is mediated by transcription factor NFkappaB. By using the specific proteasome inhibitor lactacystin to examine IkappaB degradation, we observed a paradoxical increase in lipopolysaccharide- and cytokine-dependent NOS2 expression at low concentrations or when lactacystin was added subsequent to cytokines. Lactacystin reduced the initial accumulation of NOS2 mRNA but reduced its subsequent decrease. Lactacystin increased NOS2 promoter activation after 24 h, but not after 4 h, and similarly prevented initial NFkappaB activation and at later times caused NFkappaB reactivation. Lactacystin reduced initial degradation of IkappaB-alpha and IkappaB-beta, however, at later times selectively increased IkappaB-beta, which was predominantly non-phosphorylated. Expression of full-length rat IkappaB beta, but not a carboxyl-terminal truncated form, inhibited NOS2 induction and potentiation by lactacystin. Lactacystin increased IkappaB-beta expression in the absence of NOS2 inducers, as well as expression of heat shock protein 70, and the heat shock response due to hyperthermia increased IkappaB-beta expression. These results suggest that IkappaB-beta contributes to persistent NFkappaB activation and NOS2 expression in glial cells, that IkappaB-beta is a stress protein inducible by hyperthermia or proteasome inhibitors, and that delayed addition of proteasome inhibitors can have stimulatory rather than inhibitory actions. PMID- 10827093 TI - Calcium and fos involvement in brain-derived Ca(2+)-binding protein (S100) dependent apoptosis in rat phaeochromocytoma cells. AB - Brain-derived calcium-binding protein S100 induces apoptosis in a significant fraction of rat phaeochromocytoma (PC12) cells. We used single cell techniques (patch clamp, videomicroscopy and immunocytochemistry) to clarify some of the specific aspects of S100-induced apoptosis, the modality(ies) of early intracellular Ca2+ concentration increase and the expression of some classes of genes (c-fos, c-jun, bax, bcl-x, p-15, p-21) known to be implicated in apoptosis of different cells. The results show that S100: (1) causes an increase of [Ca2+]i due to an increased conductance of L-type Ca2+ channels; (2) induces a sustained increase of the Fos levels which is evident since the first time point tested (3 h) and remains elevated until to the last time point (72 h). All these data suggest that S100-derived apoptosis in PC12 cells may be the consequence of a system involving an increase in L-type Ca2+ channel conductance with consequent [Ca2+]i increase which up-regulates, directly or indirectly, the expression of Fos. PMID- 10827094 TI - Pancreatic exocrine secretions as a source of luminal polyamines in pigs. AB - The goal of the present study was twofold: (1) to detect the possible storage of dietary polyamines (PAs) in various tissues and (2) to investigate the role of dietary PAs in the differentiation of the pig intestinal epithelium. A first experimental series was designed to assess the accumulation of either milk PAs (mostly spermidine) or orally administered spermine (SPM) in piglet red blood cells (RBCs) and plasma, a preliminary stage in their distribution to growing and storage organs. Though PA concentrations of piglet RBCs and plasma were generally significantly higher than their sow counterparts, our experimental conditions failed to demonstrate that this increase could stem from ingested PAs. A second experimental series dealt with the determination of disaccharidase specific activities in proximal and distal parts of piglet gut on the 26th and 29th days after birth (preweaning time). In agreement with observations made previously on rat pups, we observed an increase in maltase specific activity (SA) at the end of the suckling period (the observed increase in sucrase SA was not significant). However, orally administered SPM did not affect this activity. Compared to the constant protein concentrations observed in both parts of the gut, the pancreatic protein content decreased sharply between the 26th and 29th postnatal days. At the same time pancreatic concentrations of spermidine (SPD) also decreased, suggesting that some pancreatic PAs were released as the organ secreted its proteins. In accordance with this hypothesis, we recorded SPM and SPD in pancreatic juice. The increases in PA concentrations seemed to follow the protein secretion pattern (i.e. PA concentrations reached a maximal value when the protein concentration was highest). The presence of PAs in pancreatic juice could be indicative of a control mechanism exerted by the pancreas on PA-induced growth and differentiation of porcine intestinal epithelium. PMID- 10827095 TI - Do we need managers? PMID- 10827096 TI - Creating an adult learning culture through practice development. AB - The development of a learning culture is becoming a dominant theme in the strategic plans of health care organisations. This is arising through a drive to improve standards of practice, bridge the perceived theory-practice gap and create means of integrating learning with practice. There have been many initiatives to create such a change, including continuous professional development, reflective practice, clinical supervision and work based learning. This paper presents an account of a practice development strategy that aimed to create a learning culture as a sub-element of the overall programme of work. Working with individual project leaders, the intention was to shift the emphasis away from classroom based education, to learning at and from work. This was achieved through a combination of action research, the application of adult learning theory and facilitation. The paper describes the context of the development strategy, the facilitation processes adopted including the theoretical underpinnings and some 'tentative' outcomes achieved. PMID- 10827097 TI - Problem-based learning within health professional education. What is the role of the lecturer? A review of the literature. AB - With the increasing popularity of using problem-based learning (PBL) within health professional curricula, it could be argued that the health lecturer's role in education is changing. As a lecturer, I have only recently become involved in using PBL. With increasing exposure to the process and through reviewing the literature, I have come to realise that the role of the lecturer is fraught with difficulty. The literature is often conflicting with PBL meaning different things to different people (Barrows 1986). It provides no consistent guidelines as to how the lecturer should adapt to undertake this new role. This article explores the issues around the role of the lecturer within PBL and through reviewing the literature, investigates the level of intervention the lecturer should provide when students are undertaking the PBL process. Suggestions will be made to 'facilitate' the lecturer into facilitating an effective teaching strategy. PMID- 10827098 TI - Collage work as a medium for guided reflection in the clinical supervision relationship. AB - This paper identifies the value of one form of art as a means of enhancing communication and developing self expression. The medium of magazine picture collage is described with explanation of its application in the clinical supervision setting where it is used as a trigger for reflective questioning to clarify issues, examine perceptions and facilitate learning from experience. The assembly of magazine pictures as a collage allows metaphorical representation of events and influences through which the supervisee can 'see' their situation from different perspectives. This medium is used as a starting point for discussion in the supervision setting where reflective questioning by the supervisor is framed around the collage images in order to clarify relationships of the issues presented and the personal meanings and values of these to the supervisee. This method is offered as a means of supporting existing systems of supervision not a replacement for these and is perceived to be of particular value in situations where facilitation of expression by means other than dialogue alone is felt to be required in order to move the supervision process forward and enhance personal learning from this. PMID- 10827099 TI - A study of the utilization of research in practice and the influence of education. AB - This paper reports part of a multi-phase study which aimed to investigate the extent to which nurses utilize research and to identify factors associated with research utilization. The findings presented examine the influence of education upon research utilization. Firstly, a survey of registered nurses working in general medical and surgical wards in Scotland was conducted. 680/936 (72.6%) nurses returned self-report questionnaires to measure the level of utilization of 14 research based practices and assess the presence of potential influencing factors. This was then followed up through interviews with a sub-sample of nurses. An association was found between a higher educational level and research utilization. The nurses reported that in courses as opposed to study days, they were expected to engage in study and read and complete course work whereas attendance at study days could be an entirely passive experience and was often more of a morale booster. Nurses who read at least one journal regularly, had had more study leave, or had attended research courses also had a higher level of research utilization. PMID- 10827100 TI - The patient as 'teacher': learning in the care of elderly persons with dementia. AB - In 1996 HM Queen Silvia of Sweden started a non-governmental education programme with an integrated day-care unit devoted to elderly persons with dementia. A total of 18 Licensed Practical Nurses (LPN) from various parts of Sweden took part in the year theoretical and practical education in dementia care. The purpose was to develop specialized skills in the particular field and more generally to develop mentor capabilities. The aim of the study was to examine the experiences of the trainees, gained from following a single patient during their entire practical training period in the school's integrated day-care unit. The study was based on a combination of participant observations, interviews, diaries and recorded data and used an ethnographic approach. The results showed that themes such as 'personal guide', 'creating a relationship', 'reducing the working pace' and 'investigative mealtimes' were of major importance for the trainees' learning. In their role as 'personal guides', the trainees fell into six categories with different educational focuses. In their relationships with the patients, the trainees were able to form their own impressions of the patients' present and former lives. By training their ability to adapt the pace at which they worked, they had time to observe symptoms and be aware of the patients' needs, as well as the patients' signs of appreciation. The results also indicated that 'investigative mealtimes' can be an important element in the trainee's education in dementia care. PMID- 10827101 TI - Age and 'type' of domain specific entry qualifications as predictors of student nurses' performance in biological, social and behavioural sciences in nursing assessments. AB - The present study explored the effects of age and 'type' of entry qualifications in psychology, sociology and biology on student performance in 'the psychological, sociological and biological perspectives in nursing' module assessments, respectively. Data from 222 students undertaking 'the pre registration diploma in nursing' programme at a university in the north West of England were analysed. The study found no significant differences in performance among those students with GCSE 'O' level, those with access and those without any type of domain specific qualifications. However, student age significantly predicted performance, with such performances found to be highly consistent across the three modules. The 'non-mature' students (aged < 20 years) were identified in the study as being at risk in terms of academic performance whilst the 'very mature' students (aged > 34 years) were found to predict better overall performance. The findings suggest that, paper qualifications such as GCSE O.level, GNVQ or BTEC in psychology, sociology or biology should not be relied upon as predictors of academic performance in their related nursing modules when selecting potential nurses. On the basis of the findings and their probable explanations, it seems possible that the entry gate to nursing can further be widened by giving more credit to the older applicant who has, for example, achieved NVQ at level 2 or 3. However, such flexibility in nurse selection will have to be matched with equal flexibility in the pre-registration diploma in nursing curriculum which at present fails to recognize 'at risk' groups. The findings and the implications for nurse education and recruitment and discussed with support from a growing number of studies investigating student approaches to studying and learning. PMID- 10827102 TI - The nature and purpose of the role of the nurse lecturer in practice settings. AB - This paper examines the role of lecturers in nursing in pre-registration education. It focuses on the nature and purpose of the nurse lecturer's contribution in the practice setting, with particular emphasis on the issues surrounding clinical credibility. This is particularly pertinent in the light of current recommendations, which emphasise the importance of clinical learning in pre-registration education programmes. The purpose of the lecturer's role in clinical practice settings is ill defined. This lack of clear consensus regarding the expected outcomes for lecturers (in practice), leads to difficulty outlining what they should do in practice settings. Although lecturers accept that they have an important part to play in maximizing the learning opportunities for students in both university and practice settings, they are less clear about how this should be achieved in the latter. This paper argues that: It is opportune to examine and realign the lecturers contribution in practice settings given that there has been a shift in the responsibility for clinical learning; nurse education is now embedded in the higher education sector and there is a need for a greater emphasis on practice development. Clinical credibility for lecturers is about the development of nursing practice through education which is not always achieved by 'hands on' care. For example, assisting nurses in a ward area to develop expertise in evidence based practice may not involve 'hands on' care giving but does involve being conversant with current research and practice issues. The lecturer's expertise in practice settings is in teaching and facilitating learning, rather than direct care giving. No one common model for practice may be either feasible or desirable. However, it is important that nurse lecturers do not follow a particular approach because the debate about the nurse lecturer's role in practice settings fails to acknowledge the strengths and weaknesses of each model. It is important that the approach is based on sound rationale. PMID- 10827103 TI - Can educational methods influence the popularity of psychiatric nursing? AB - The ongoing difficulty of attracting sufficient numbers of undergraduate nursing students to a career in psychiatric nursing has been acknowledged. Limited available research suggests that education can produce a more positive view of psychiatric nursing. Despite these findings, research into the impact of educational methods upon the popularity of psychiatric nursing has not been undertaken. This paper reports the findings of a quasi-experimental research project, which examined the relationship between the study of, and the popularity of, psychiatric nursing. The results propose that through undertaking the psychiatric nursing program, psychiatric nursing as a future career option became more popular. The greater significance of the results of this study than had been found in previous studies suggests that problem-based learning is an educational method capable of influencing the development of more positive attitudes. PMID- 10827104 TI - Essential clinical skills for beginning registered nurses. AB - A research project investigating the identification of essential clinical skills for registered nurses in their first year of practice was undertaken in a number of Sydney metropolitan hospitals in Australia. Using an adaptation of the Williams & Brammer (1996) study, 206 beginning registered nurses were surveyed to determine the clinical skills they performed frequently in their daily routine, and those they considered essential. One hundred and thirty two questionnaires were returned and the responses were analysed for frequency of clinical skill performance and essential rating. The clinical skills rated as most frequently performed related to universal precautions for infection control, vital sign assessment, management of intravenous therapy, administration of all forms of medication and a wide range of patient hygiene related skills. Many clinical skills rated as essential by these nurses were not rated as performed frequently, which may indicate concern for the need to draw upon less frequently performed skills during emergency situations and thus they rate these skills as essential. This study clearly identified the clinical skills rated as performed frequently by beginning registered nurses and identified those clinical skills deemed essential to beginning nursing practice by novice nurses. A larger sample size would strengthen the reliability of these findings. PMID- 10827105 TI - A short tandem repeat-based phylogeny for the human Y chromosome. AB - Human Y-chromosomal short tandem repeat (STR) data provide a potential model system for the understanding of autosomal STR mutations in humans and other species. Yet, the reconstruction of STR evolution is rarely attempted, because of the absence of an appropriate methodology. We here develop and validate a phylogenetic-network approach. We have typed 256 Y chromosomes of indigenous descent from Africa, Asia, Europe, Australia, and highland Papua New Guinea, for the STR loci DYS19, DXYS156Y, DYS389, DYS390, DYS392, and DYS393, as well as for five ancient biallelic mutation events: two poly (A) length variants associated with the YAP insertion, two independent SRY-1532 mutations, and the 92R7 mutation. We have used our previously published pedigree data from 11,000 paternity-tested autosomal STR-allele transfers to produce a two-class weighting system for the Y-STR loci that is based on locus lengths and motif lengths. Reduced-median-network analysis yields a phylogeny that is independently supported by the five biallelic mutations, with an error of 6%. We find the earliest branch in our African San (Bushmen) sample. Assuming an age of 20,000 years for the Native American DYS199 T mutation, we estimate a mutation rate of 2.6x10-4 mutations/20 years for slowly mutating Y STRs, approximately 10-fold slower than the published average pedigree rate. PMID- 10827106 TI - Aicardi-Goutieres syndrome displays genetic heterogeneity with one locus (AGS1) on chromosome 3p21. AB - We have studied 23 children from 13 families with a clinical diagnosis of Aicardi Goutieres syndrome. Affected individuals had developed an early-onset progressive encephalopathy that was characterized by a normal head circumference at birth, basal ganglia calcification, negative viral studies, and abnormalities of cerebrospinal fluid comprising either raised white cell counts and/or raised levels of interferon-alpha. By means of genomewide linkage analysis, a maximum heterogeneity LOD score of 5.28 was reached at marker D3S3563, with alpha=.48, where alpha is the proportion of families showing linkage. Our data suggest the existence of locus heterogeneity in Aicardi-Goutieres syndrome and highlight potential difficulties in the differentiation of this condition from pseudo-TORCH (toxoplasmosis, rubella, cytomegalovirus, and herpes simplex virus types 1 and 2) syndrome. PMID- 10827107 TI - Association mapping in structured populations. AB - The use, in association studies, of the forthcoming dense genomewide collection of single-nucleotide polymorphisms (SNPs) has been heralded as a potential breakthrough in the study of the genetic basis of common complex disorders. A serious problem with association mapping is that population structure can lead to spurious associations between a candidate marker and a phenotype. One common solution has been to abandon case-control studies in favor of family-based tests of association, such as the transmission/disequilibrium test (TDT), but this comes at a considerable cost in the need to collect DNA from close relatives of affected individuals. In this article we describe a novel, statistically valid, method for case-control association studies in structured populations. Our method uses a set of unlinked genetic markers to infer details of population structure, and to estimate the ancestry of sampled individuals, before using this information to test for associations within subpopulations. It provides power comparable with the TDT in many settings and may substantially outperform it if there are conflicting associations in different subpopulations. PMID- 10827108 TI - Additional copies of the proteolipid protein gene causing Pelizaeus-Merzbacher disease arise by separate integration into the X chromosome. AB - The proteolipid protein gene (PLP) is normally present at chromosome Xq22. Mutations and duplications of this gene are associated with Pelizaeus-Merzbacher disease (PMD). Here we describe two new families in which males affected with PMD were found to have a copy of PLP on the short arm of the X chromosome, in addition to a normal copy on Xq22. In the first family, the extra copy was first detected by the presence of heterozygosity of the AhaII dimorphism within the PLP gene. The results of FISH analysis showed an additional copy of PLP in Xp22.1, although no chromosomal rearrangements could be detected by standard karyotype analysis. Another three affected males from the family had similar findings. In a second unrelated family with signs of PMD, cytogenetic analysis showed a pericentric inversion of the X chromosome. In the inv(X) carried by several affected family members, FISH showed PLP signals at Xp11.4 and Xq22. A third family has previously been reported, in which affected members had an extra copy of the PLP gene detected at Xq26 in a chromosome with an otherwise normal banding pattern. The identification of three separate families in which PLP is duplicated at a noncontiguous site suggests that such duplications could be a relatively common but previously undetected cause of genetic disorders. PMID- 10827109 TI - The exon 13 duplication in the BRCA1 gene is a founder mutation present in geographically diverse populations. The BRCA1 Exon 13 Duplication Screening Group. AB - Recently, a 6-kb duplication of exon 13, which creates a frameshift in the coding sequence of the BRCA1 gene, has been described in three unrelated U.S. families of European ancestry and in one Portuguese family. Here, our goal was to estimate the frequency and geographic diversity of carriers of this duplication. To do this, a collaborative screening study was set up that involved 39 institutions from 19 countries and included 3,580 unrelated individuals with a family history of the disease and 934 early-onset breast and/or ovarian cancer cases. A total of 11 additional families carrying this mutation were identified in Australia (1), Belgium (1), Canada (1), Great Britain (6), and the United States (2). Haplotyping showed that they are likely to derive from a common ancestor, possibly of northern British origin. Our results demonstrate that it is strongly advisable, for laboratories carrying out screening either in English-speaking countries or in countries with historical links with Britain, to include within their BRCA1 screening protocols the polymerase chain reaction-based assay described in this report. PMID- 10827110 TI - Breaking away from home. PMID- 10827111 TI - Remote sensing and human health: new sensors and new opportunities. AB - Since the launch of Landsat-1 28 years ago, remotely sensed data have been used to map features on the earth's surface. An increasing number of health studies have used remotely sensed data for monitoring, surveillance, or risk mapping, particularly of vector-borne diseases. Nearly all studies used data from Landsat, the French Systeme Pour l'Observation de la Terre, and the National Oceanic and Atmospheric Administration's Advanced Very High Resolution Radiometer. New sensor systems are in orbit, or soon to be launched, whose data may prove useful for characterizing and monitoring the spatial and temporal patterns of infectious diseases. Increased computing power and spatial modeling capabilities of geographic information systems could extend the use of remote sensing beyond the research community into operational disease surveillance and control. This article illustrates how remotely sensed data have been used in health applications and assesses earth-observing satellites that could detect and map environmental variables related to the distribution of vector-borne and other diseases. PMID- 10827112 TI - A dynamic transmission model for predicting trends in Helicobacter pylori and associated diseases in the United States. AB - To assess the benefits of intervention programs against Helicobacter pylori infection, we estimated the baseline curves of its incidence and prevalence. We developed a mathematical (compartmental) model of the intrinsic dynamics of H. pylori, which represents the natural history of infection and disease progression. Our model divided the population according to age, infection status, and clinical state. Case-patients were followed from birth to death. A proportion of the population acquired H. pylori infection and became ill with gastritis, duodenal ulcer, chronic atrophic gastritis, or gastric cancer. We simulated the change in transmissibility consistent with the incidence of gastric cancer and duodenal ulcer over time, as well as current H. pylori prevalence. In the United States, transmissibility of H. pylori has decreased to values so low that, should this trend continue, the organism will disappear from the population without targeted intervention; this process, however, will take more than a century. PMID- 10827113 TI - Using remotely sensed data to identify areas at risk for hantavirus pulmonary syndrome. AB - The 1993 U.S. hantavirus pulmonary syndrome (HPS) outbreak was attributed to environmental conditions and increased rodent populations caused by unusual weather in 1991- 92. In a case-control study to test this hypothesis, we estimated precipitation at 28 HPS and 170 control sites during the springs of 1992 and 1993 and compared it with precipitation during the previous 6 years by using rainfall patterns at 196 weather stations. We also used elevation data and Landsat Thematic Mapper satellite imagery collected the year before the outbreak to estimate HPS risk by logistic regression analysis. Rainfall at case sites was not higher during 1992-93 than in previous years. However, elevation, as well as satellite data, showed association between environmental conditions and HPS risk the following year. Repeated analysis using satellite imagery from 1995 showed substantial decrease in medium- to high-risk areas. Only one case of HPS was identified in 1996. PMID- 10827114 TI - Remote sensing and geographic information systems: charting Sin Nombre virus infections in deer mice. AB - We tested environmental data from remote sensing and geographic information system maps as indicators of Sin Nombre virus (SNV) infections in deer mouse (Peromyscus maniculatus) populations in the Walker River Basin, Nevada and California. We determined by serologic testing the presence of SNV infections in deer mice from 144 field sites. We used remote sensing and geographic information systems data to characterize the vegetation type and density, elevation, slope, and hydrologic features of each site. The data retroactively predicted infection status of deer mice with up to 80% accuracy. If models of SNV temporal dynamics can be integrated with baseline spatial models, human risk for infection may be assessed with reasonable accuracy. PMID- 10827115 TI - Potential exposure to Australian bat lyssavirus, Queensland, 1996-1999. AB - Two human deaths caused by Australian bat lyssavirus (ABL) infection have been reported since 1996. Information was obtained from 205 persons (mostly adults from south Brisbane and the South Coast of Queensland), who reported potential ABL exposure to the Brisbane Southside Public Health Unit from November 1,1996, to January 31, 1999. Volunteer animal handlers accounted for 39% of potential exposures, their family members for 12%, professional animal handlers for 14%, community members who intentionally handled bats for 31%, and community members with contacts initiated by bats for 4%. The prevalence of Lyssavirus detected by fluorescent antibody test in 366 sick, injured, or orphaned bats from the area was 6%. Sequelae of exposure, including the requirement for expensive postexposure prophylaxis, may be reduced by educating bat handlers and the public of the risks involved in handling Australian bats. PMID- 10827117 TI - Rhinosporidium seeberi: a human pathogen from a novel group of aquatic protistan parasites. AB - Rhinosporidium seeberi, a microorganism that can infect the mucosal surfaces of humans and animals, has been classified as a fungus on the basis of morphologic and histochemical characteristics. Using consensus polymerase chain reaction (PCR), we amplified a portion of the R. seeberi 18S rRNA gene directly from infected tissue. Analysis of the aligned sequence and inference of phylogenetic relationships showed that R. seeberi is a protist from a novel clade of parasites that infect fish and amphibians. Fluorescence in situ hybridization and R. seeberi- specific PCR showed that this unique 18S rRNA sequence is also present in other tissues infected with R. seeberi. Our data support the R. seeberi phylogeny recently suggested by another group. R. seeberi is not a classic fungus, but rather the first known human pathogen from the DRIPs clade, a novel clade of aquatic protistan parasites (Ichthyosporea). PMID- 10827116 TI - Genetic variation in Pneumocystis carinii isolates from different geographic regions: implications for transmission. AB - To study transmission patterns of Pneumocystis carinii pneumonia (PCP) in persons with AIDS, we evaluated P. carinii isolates from patients in five U.S. cities for variation at two independent genetic loci, the mitochondrial large subunit rRNA and dihydropteroate synthase. Fourteen unique multilocus genotypes were observed in 191 isolates that were examined at both loci. Mixed infections, accounting for 17.8% of cases, were associated with primary PCP. Genotype frequency distribution patterns varied by patients' place of diagnosis but not by place of birth. Genetic variation at the two loci suggests three probable characteristics of transmission: that most cases of PCP do not result from infections acquired early in life, that infections are actively acquired from a relatively common source (humans or the environment), and that humans, while not necessarily involved in direct infection of other humans, are nevertheless important in the transmission cycle of P. carinii f. sp. hominis. PMID- 10827118 TI - High prevalence of penicillin-nonsusceptible Streptococcus pneumoniae at a community hospital in Oklahoma. AB - During 1997, Oklahoma City's Hospital A reported penicillin-nonsusceptible Streptococcus pneumoniae in almost 67% of isolates. To confirm this finding, all Hospital A S. pneumoniae isolates from October 23, 1997, through February 19, 1998, were tested for antibiotic susceptibility and repeat-tested at two other hospital laboratories. Medical records of Hospital A patients with invasive S. pneumoniae infections during 1994 through 1997 were also reviewed. These data were compared with 1998 statewide sentinel hospital surveillance data for invasive S. pneumoniae. Of 48 S. pneumoniae isolates from Hospital A during October 23, 1997, through February 19, 1998, 31 (65%) were penicillin nonsusceptible S. pneumoniae, and 23 (48%) were highly penicillin resistant. Similar prevalences were confirmed at the other hospital laboratories; however, significant interlaboratory differences were noted in the determination of third generation cephalosporin susceptibility. During 1994 through 1997, a trend toward increasing penicillin nonsusceptibility (p <0.05) was noted among S. pneumoniae isolates from nursing home patients. During 1998, 85 (30%) of 282 invasive isolates reported to the state surveillance system were penicillin-nonsusceptible S. pneumoniae; 33 (12%) were highly resistant. The increase in resistance observed is notable; the interlaboratory discrepancies are unexplained. To respond, a vaccination program was implemented at Hospital A, and vaccination efforts were initiated at nursing homes. PMID- 10827119 TI - Rickettsia mongolotimonae: a rare pathogen in France. AB - We report a second case of laboratory-confirmed infection caused by Rickettsia mongolotimonae in Marseille, France. This rickettsiosis may represent a new clinical entity; moreover, its geographic distribution may be broader than previously documented. This pathogen should be systematically considered in the differential diagnosis of atypical rickettsioses, especially rashless fevers with lymphangitis and lymphadenopathy, in southern France and perhaps elsewhere. PMID- 10827120 TI - Costs and benefits of a subtype-specific surveillance system for identifying Escherichia coli O157:H7 outbreaks. AB - We assessed the societal costs and benefits of a subtype-specific surveillance system for identifying outbreak-associated Escherichia coli O157:H7 infections. Using data from Colorado, we estimated that if it averted five cases annually, the system would recover all its costs. PMID- 10827121 TI - Dengue epidemic in Belem, Para, Brazil, 1996-97. AB - We describe clinical and epidemiologic findings during the first epidemic of dengue fever in Belem, Para State, Brazil, in 1996-97. Of 40,237 serum samples, 17,440 (43%) were positive for dengue by virus isolation or serologic testing. No hemorrhagic cases or deaths were reported. Mycobacterium tuberculosis PMID- 10827122 TI - Mycobacterium tuberculosis Beijing genotype emerging in Vietnam. AB - To assess whether the Mycobacterium tuberculosis Beijing genotype is emerging in Vietnam, we analyzed 563 isolates from new cases by spoligotyping and examined the association between the genotype and age, resistance, and BCG vaccination status. Three hundred one (54%) patients were infected with Beijing genotype strains. The genotype was associated with younger age (and hence with active transmission) and with isoniazid and streptomycin resistance, but not with BCG vaccination. PMID- 10827123 TI - Bartonella spp. isolated from wild and domestic ruminants in North America. AB - Bartonella species were isolated from 49% of 128 cattle from California and Oklahoma, 90% of 42 mule deer from California, and 15% of 100 elk from California and Oregon. Isolates from all 63 cattle, 14 deer, and 1 elk had the same polymerase chain reaction/restriction fragment length polymorphism profiles. Our findings indicate potential for inter- and intraspecies transmission among ruminants, as well as risk that these Bartonella spp. could act as zoonotic agents. PMID- 10827124 TI - Malaria and global warming in perspective? PMID- 10827125 TI - Serologic evidence of human monocytic and granulocytic ehrlichiosis in Israel. PMID- 10827127 TI - Death receptors, intimal disease, and gene therapy: are therapies that modify cell fate moving too Fas? PMID- 10827128 TI - Genetic susceptibility to atherosclerosis: insights from mice. PMID- 10827129 TI - Calcium waves: physiological relevance in cardiac function. PMID- 10827130 TI - Small guanine nucleotide-binding proteins and myocardial hypertrophy. AB - The small (21 kDa) guanine nucleotide-binding protein (small G protein) superfamily comprises 5 subfamilies (Ras, Rho, ADP ribosylation factors [ARFs], Rab, and Ran) that act as molecular switches to regulate numerous cellular responses. Cardiac myocyte hypertrophy is associated with cell growth and changes in the cytoskeleton and myofibrillar apparatus. In other cells, the Ras subfamily regulates cell growth whereas the Rho subfamily (RhoA, Rac1, and Cdc42) regulates cell morphology. Thus, the involvement of small G proteins in hypertrophy has become an area of significant interest. Hearts from transgenic mice expressing activated Ras develop features consistent with hypertrophy, whereas mice overexpressing RhoA develop lethal heart failure. In isolated neonatal rat cardiac myocytes, transfection or infection with activated Ras, RhoA, or Rac1 induces many of the features of hypertrophy. We discuss the mechanisms of activation of the small G proteins and the downstream signaling pathways involved. The latter may include protein kinases, particularly the mitogen activated or Rho-activated protein kinases. We conclude that although there is significant evidence implicating Ras, RhoA, and Rac1 in hypertrophy, the mechanisms are not fully understood. PMID- 10827131 TI - Cardiovascular overexpression of transforming growth factor-beta(1) causes abnormal yolk sac vasculogenesis and early embryonic death. AB - Transforming growth factor-beta(1) (TGF-beta(1)) is expressed in the adult and embryonic vasculature; however, the biological consequences of increased vascular TGF-beta(1) expression remain controversial. To establish an experimental setting for investigating the role of increased TGF-beta(1) in vascular development and disease, we generated transgenic mice in which a cDNA encoding a constitutively active form of TGF-beta(1) is expressed from the SM22alpha promoter. This promoter fragment directs transgene expression to smooth muscle cells of large arteries in late-term embryos and postnatal mice. We confirmed the anticipated pattern of SM22alpha-directed transgene expression (heart, somites, and vasculature of the embryo and yolk sac) in embryos carrying an SM22alpha-beta galactosidase transgene. SM22alpha- beta-galactosidase transgenic mice were born at the expected frequency (13%); however, nearly all SM22alpha-TGF-beta(1) transgenic mice died before E11.5. SM22alpha-TGF-beta(1) transgenic embryos identified at E8.5 to E10.5 had growth retardation and both gross and microscopic abnormalities of the yolk sac vasculature. Overexpression of TGF-beta(1) from the SM22alpha promoter is lethal at E8.5 to E10.5, most likely because of yolk sac insufficiency. Investigation of the consequences of increased vascular TGF beta(1) expression in adults may require a conditional transgenic approach. Moreover, because the SM22alpha promoter drives transgene expression in the yolk sac vasculature at a time when embryonic survival is dependent on yolk sac function, use of the SM22alpha promoter to drive expression of "vasculoactive" transgenes may be particularly likely to cause embryonic death. PMID- 10827132 TI - Immunoneutralization of glycoprotein Ibalpha attenuates endotoxin-induced interactions of platelets and leukocytes with rat venular endothelium in vivo. AB - This study aimed to examine molecular mechanisms for endotoxin-induced adhesive changes in platelets in vivo. Platelets labeled with carboxyfluorescein diacetate succinimidyl ester were visualized in rat mesenteric venules through intravital microscopy assisted by a high-speed fluorescence video imager at 1000 frames per second or by a normal-speed intensifier under monitoring of erythrocyte velocity. Leukocyte rolling was examined by normal-speed transmission video images. The velocity of platelets traveling along the centerline of venules followed that of erythrocytes, whereas that measured at the periendothelial space was significantly smaller than the erythrocyte velocity; a majority of these cells exhibited transient but notable rolling with endothelium. Administration of endotoxin increased the density of periendothelial platelets and reduced the rolling velocities of platelets and leukocytes in venules: All events were attenuated by anti-rat P-selectin monoclonal antibody s789G or by anti-human glycoprotein (GP) Ibalpha monoclonal antibody GUR83/35, which blocks ristocetin induced aggregation of rat platelets. Isolated rat platelets injected into endotoxin-pretreated rats were able to roll on the venules. This event was attenuated by pretreatment of platelets in vitro with GUR83/35 but not with s789G, suggesting involvement of endothelial P-selectin and platelet GP Ibalpha in the endotoxin-induced responses. Furthermore, isolated human platelets showed similar rolling interactions with endotoxin-preexposed rat venules, and pretreatment of the platelets with GUR83/35, but not with s789G, significantly reduced such interactions. Our results provide the first evidence for involvement of GP Ibalpha in endotoxin-induced microvascular rolling of platelets and leukocytes, and this system serves as a potentially useful tool to examine GP Ibalpha-associated function of human platelets in vivo. PMID- 10827133 TI - Sensitivity to Fas-mediated apoptosis is determined below receptor level in human vascular smooth muscle cells. AB - Despite Fas expression, many cells resist Fas-induced apoptosis. Although differences in surface Fas expression can explain Fas resistance, multiple proteins below receptor level also inhibit Fas-induced apoptosis. To examine the mechanism of Fas resistance, we studied Fas-induced apoptosis in human medial vascular smooth muscle cells (VSMCs) from healthy coronary arteries. VSMCs showed marked heterogeneity to Fas-induced apoptosis, exhibiting both Fas-resistant (98.1+/-2.3% viable, n = 4, P = NS) and Fas-sensitive (31.3+/-2.6% viable, n = 3, P<0.01) cells. Fas-resistant VSMCs expressed surface Fas and could recruit RIP, indicating that functional receptor complexes were formed. However, Fas-resistant cells showed reduced expression of FADD, Fas ligand, and caspases 3, 7, and 8 and increased expression of FLIP and c-IAP-1. Fas-induced apoptosis was associated with cleavage of caspase 3 and blocked by inhibitors of caspase 3 or 8 but not caspase 1, 6, or 7. Selective inhibition of caspase 3 or 8 by antisense transfection inhibited Fas-induced apoptosis, but their reexpression could not rescue the Fas-resistant phenotype. In vivo, medial VSMCs showed marked heterogeneity of expression of caspase 3. We conclude that Fas sensitivity is determined not only by expression of surface Fas but by differential expression of Fas-signaling proteins below receptor level. Subpopulations of cells within the same tissue have different sensitivities to apoptosis, determined by expression of specific death-signaling proteins. PMID- 10827134 TI - Coupling function of endogenous alpha(1)- and beta-adrenergic receptors in mouse cardiomyocytes. AB - Genetically altered mouse models constitute unique systems to delineate the role of adrenergic receptor (AR) signaling mechanisms as modulators of cardiomyocyte function. The interpretation of results from these models depends on knowledge of the signaling properties of endogenous ARs in mouse cardiomyocytes. In the present study, we identify for the first time several defects in AR signaling in cardiomyocytes cultured from mouse ventricles. beta(1)-ARs induce robust increases in cAMP accumulation and the amplitude of the calcium and cell motion transients in mouse cardiomyocytes. Selective beta(2)-AR stimulation increases the amplitude of calcium and motion transients, with only a trivial rise in cAMP accumulation in comparison. beta(2)-AR responses are not influenced by pertussis toxin in cultured mouse cardiomyocytes. alpha(1)-ARs fail to activate phospholipase C, the extracellular signal-regulated protein kinase, p38-MAPK, or stimulate hypertrophy in mouse cardiomyocytes. Control experiments establish that this is not due to a lesion in distal elements in the signaling machinery, because these responses are induced by protease-activated receptor-1 agonists and phospholipase C is activated by Pasteurella multocida toxin (a G(q) alpha-subunit agonist). Surprisingly, norepinephrine activates p38-MAPK via beta-ARs in mouse cardiomyocytes, but beta-AR activation of p38-MAPK alone is not sufficient to induce cardiomyocyte hypertrophy. Collectively, these results identify a generalized defect in alpha(1)-AR signaling and a defect in beta(2)-AR linkage to cAMP (although not to an inotropic response) in cultured mouse cardiomyocytes. These naturally occurring vagaries in AR signaling in mouse cardiomyocytes provide informative insights into the requirements for hypertrophic signaling and impact on the value of mouse cardiomyocytes as a reconstitution system to investigate AR signaling in the heart. PMID- 10827135 TI - Signaling properties and functions of two distinct cardiomyocyte protease activated receptors. AB - Previous studies have established that cardiomyocytes express protease-activated receptor (PAR)-1, a high-affinity receptor for thrombin, which is also activated by the tethered-ligand domain sequence (SFLLRN) and which promotes inositol trisphosphate accumulation, stimulates extracellular signal-regulated protein kinase, and modulates contractile function. A single previous report identified PAR-1 as a hypertrophic stimulus, but there have been no subsequent investigations of the mechanism. This study reveals the coexpression of PAR-1 and PAR-2 (a second PAR, which is activated by trypsin/tryptase but not thrombin) by Northern blot analysis and compares their signaling properties in neonatal rat ventricular cardiomyocytes. SFLLRN and SLIGRL (an agonist peptide for PAR-2) promote inositol trisphosphate accumulation, stimulate mitogen-activated protein kinases (extracellular signal-regulated protein kinase and p38-mitogen-activated protein kinase), elevate calcium concentration, and increase spontaneous automaticity. SFLLRN (but not SLIGRL) also activates c-Jun NH(2)-terminal kinase and AKT. In keeping with their linkage to pathways that have been associated with growth and/or survival, SFLLRN and SLIGRL both induce hypertrophy. However, PAR agonists promote cell elongation, a morphology that is distinct from the uniform increase in cell dimension induced by alpha(1)-adrenergic receptor activation. These studies provide novel evidence that cardiomyocytes coexpress 2 functional PARs, which link to a common set of signals that culminate in changes in contractile function and hypertrophic growth. PAR actions may assume clinical importance in the border zone surrounding an infarction, where local proteolysis of PARs by serine proteases generated during inflammatory or thrombogenic pathways would elevate calcium concentration (setting the stage for arrhythmias), promote hypertrophic growth, and/or influence cardiomyocyte survival. PMID- 10827136 TI - Effects of the renin-angiotensin system on the current I(to) in epicardial and endocardial ventricular myocytes from the canine heart. AB - The Ca(2+)-independent portion of transient outward K(+) current (I(to)) exhibits a transmural gradient in ventricle. To investigate control mechanisms for this gradient, we studied canine epicardial and endocardial ventricular myocytes with use of the whole-cell patch-clamp technique. I(to) was larger in amplitude, had a more negative voltage threshold for activation, and had a more negative midpoint of inactivation in epicardium. Recovery from inactivation was >10-fold slower in endocardium. Incubation of epicardial myocytes with angiotensin II for 2 to 52 hours altered I(to) to resemble unincubated endocardium and reduced the amplitude of the phase 1 notch of the action potential. In contrast, incubation of endocardial myocytes with losartan for 2 to 52 hours altered I(to) to resemble unincubated epicardium and induced a phase 1 notch in the action potential. With RNase protection assays, we determined that incubations with angiotensin II or losartan did not alter mRNA levels for either Kv4.3 or Kv1.4; thus, a change in the alpha subunit for I(to) is unlikely to be responsible. To test whether posttranslational modification produced the effects of angiotensin II, we coexpressed Kv4.3 and the angiotensin II type 1a receptor in Xenopus oocytes. Incubation with angiotensin II increased the time constant for recovery from inactivation of the expressed current by 2-fold with an incubation time constant of 3.7 hours. No effect on activation or inactivation voltage dependence was observed. These results demonstrate that the properties of I(to) in endocardium and epicardium are plastic and likely under the tonic-differing influence of the renin-angiotensin system. PMID- 10827137 TI - Enhanced cardiomyocyte DNA synthesis during myocardial hypertrophy in mice expressing a modified TSC2 transgene. AB - Tuberous sclerosis complex (TSC) is a rare genetic disorder characterized by the appearance of benign tumors in multiple organs, including the heart. Disease progression is accompanied by homozygous mutation at 1 of 2 loci (designated TSC1 or TSC2), leading to the suggestion that these genes function as tumor suppressors. In this study, we generated a series of TSC2 cDNAs in which one or more structural motifs were deleted, with the hope that expression of the modified gene product would override the growth-inhibitory activity of the endogenous TSC2 gene product. Several of the modified cDNAs enhanced growth rate, increased endocytosis, and promoted aberrant protein trafficking when expressed in NIH-3T3 cells, thereby mimicking phenotypes typical of TSC2-deficient cells. Surprisingly, targeted expression of the most potent TSC2 cDNA to the heart did not perturb cardiac development. However, the level of cardiomyocyte DNA synthesis in adult transgenic mice was elevated >35-fold during isoproterenol induced hypertrophy compared with their nontransgenic siblings. These results suggest that alteration of TSC2 gene activity in combination with beta-adrenergic stimulation can reactivate the cell cycle in a limited number of terminally differentiated adult cardiomyocytes. PMID- 10827138 TI - Determinants of atherosclerosis susceptibility in the C3H and C57BL/6 mouse model: evidence for involvement of endothelial cells but not blood cells or cholesterol metabolism. AB - Lipids, monocytes, and arterial wall cells are primary components involved in atherogenesis. Using the inbred mouse strains C57BL/6J (B6) and C3H/HeJ (C3H), which have been extensively studied as models of the genetic control of diet induced atherosclerosis, we examined which of these components determine genetic susceptibility. To test whether dietary responsiveness is involved, a congenic strain of C3H carrying an apoE-null allele (apoE(-/-)) was constructed. Although C3H.apoE(-/-) mice had higher plasma cholesterol levels, they developed much smaller lesions than their B6.apoE(-/-) counterpart on either chow or Western diets. Reciprocal bone marrow transplantation between the strains, with congenics carrying the same H-2 haplotype, was performed to examine the role of monocytes. The atherosclerosis susceptibility was not altered in the recipient mice, indicating that variations in monocyte function were not involved. Endothelial cells isolated from the aorta of B6 mice exhibited a dramatic induction of monocyte chemotactic protein-1, macrophage colony-stimulating factor, vascular cell adhesion molecule-1, and heme oxygenase-1 in response to minimally modified LDL, whereas endothelial cells from C3H mice showed little or no induction. In a set of recombinant inbred strains derived from the B6 and C3H parental strains, endothelial responses to minimally modified LDL cosegregated with aortic lesion size. These data provide strong evidence that endothelial cells, but not monocytes or plasma lipid levels, account for the difference in susceptibility to atherosclerosis between the 2 mouse strains. PMID- 10827139 TI - Contribution of caveolin protein abundance to augmented nitric oxide signaling in conscious dogs with pacing-induced heart failure. AB - Myocardial NO signaling appears elevated in heart failure (HF). Whether this results from increased NO production, induction of the high-output NO synthase (NOS)2 isoform, or changes in NOS regulatory pathways (such as caveolae) remains controversial. We tested the hypothesis that increased abundance of caveolin-3 and/or sarcolemmal caveolae contribute to increased NO signaling in pacing induced HF. Abundance of caveolin-3 (0.59+/-0.08 versus 0.29+/-0.08 arbitrary units, P = 0.01) but not caveolin-1 was increased in HF compared with control conditions, assessed by Western blot. Additionally, transmission electron microscopy revealed increased caveolae (2. 7+/-0.4 versus 1.3+/-0.3 per micrometer myocyte membrane, P<0.005). The association between caveolin-3 and NOS3 at the sarcolemma and T tubules was unchanged in HF compared with control myocytes. The impact of NOS inhibition with L-N(G)-methylarginine hydrochloride (L-NMMA) on beta-adrenergic inotropy was assessed in conscious dogs before and after HF. In control dogs, dobutamine (5 microg. kg(-1) x min(-1)) increased +dP/dt by 36+/-7%, and this was augmented to 66+/-24% by 20 mg/kg L-NMMA (P = 0.04 versus without L-NMMA, n = 8) but not affected by 10 mg/kg L-NMMA (34+/-10%, P = NS; n = 8). In HF, dobutamine +dP/dt response was depressed (P<0.001 versus control), and increased concentrations were required to match control inotropic responses (10 to 15 microg. kg(-1) x min(-1), 48+/-7%). L-NMMA enhanced +dP/dt responses similarly at 10 mg/kg (61+/-17%, P = 0.02; n = 4) and 20 mg/kg (54+/ 7%, P = 0.04; n = 7). Caveolin-3 abundance positively correlated with L-NMMA augmentation of dobutamine inotropic responses in HF (r = 0.9, P = 0.03; n = 4). Thus, in canine pacing-induced HF, expression of caveolin-3 and of sarcolemmal caveolae is increased. This increase is associated with augmented agonist stimulated NO signaling, likely via a compartmentation effect. PMID- 10827141 TI - UltraRapid communication : coexpression of connexins 40 and 43 enhances the pH sensitivityof gap junctions: A model for synergistic interactions among connexins AB - Gap junctions are formed by oligomerization of a protein called connexin. Most cells express more than one connexin isotype. Atrial myocytes, for example, coexpress connexin (Cx) 40 and Cx43. The consequence of connexin coexpression on the regulation of gap junctions is not well understood. In the present study, we show that cells coexpressing Cx40 and Cx43 are more susceptible to acidification induced uncoupling than those cells expressing only one connexin isotype. Xenopus oocytes were injected with mRNA for Cx40, Cx43, or a combination of both. Intracellular pH and junctional conductance were simultaneously measured while cells were progressively acidified by superfusion with a bicarbonate-buffered solution gassed with increasing concentrations of carbon dioxide. The data show that the pKa (ie, the pH at which junctional conductance decreased to 50% from maximum) shifted from approximately 6.7 when cells expressed only Cx40 or only Cx43 to approximately 7.0 when one of the oocytes was coexpressing both connexins. Truncation of the carboxyl terminal domains of the connexins caused the loss of pH sensitivity even after coexpression. The data are interpreted on the basis of previous studies from our laboratory that demonstrated heterodomain interactions in the regulation of Cx40 and Cx43 gap junctions. The possible implications of these findings on the regulation of native gap junctions that express both connexins remain to be determined. The full text of this article is available at http://www.circresaha.org. Web Site Feature The full-length article can be found on the World Wide Web at http://www.circresaha.org Key Words: connexin gap junctions pH(i) PMID- 10827140 TI - Three distinct types of Ca(2+) waves in Langendorff-perfused rat heart revealed by real-time confocal microscopy. AB - Although Ca(2+) waves in cardiac myocytes are regarded as arrhythmogenic substrates, their properties in the heart in situ are poorly understood. On the hypothesis that Ca(2+) waves in the heart behave diversely and some of them influence the cardiac function, we analyzed their incidence, propagation velocity, and intercellular propagation at the subepicardial myocardium of fluo 3 loaded rat whole hearts using real-time laser scanning confocal microscopy. We classified Ca(2+) waves into 3 types. In intact regions showing homogeneous Ca(2+) transients under sinus rhythm (2 mmol/L [Ca(2+)](o)), Ca(2+) waves did not occur. Under quiescence, the waves occurred sporadically (3.8 waves. min(-1) x cell(-1)), with a velocity of 84 microm/s, a decline half-time (t(1/2)) of 0.16 seconds, and rare intercellular propagation (propagation ratio <0.06) (sporadic wave). In contrast, in presumably Ca(2+)-overloaded regions showing higher fluorescent intensity (113% versus the intact regions), Ca(2+) waves occurred at 28 waves x min(-1) x cell(-1) under quiescence with a higher velocity (116 microm/s), longer decline time (t(1/2) = 0.41 second), and occasional intercellular propagation (propagation ratio = 0.23) (Ca(2+)-overloaded wave). In regions with much higher fluorescent intensity (124% versus the intact region), Ca(2+) waves occurred with a high incidence (133 waves x min(-1) x cell(-1)) and little intercellular propagation (agonal wave). We conclude that the spatiotemporal properties of Ca(2+) waves in the heart are diverse and modulated by the Ca(2+)-loading state. The sporadic waves would not affect cardiac function, but prevalent Ca(2+)-overloaded and agonal waves may induce contractile failure and arrhythmias. PMID- 10827142 TI - Coexpression of connexins 40 and 43 enhances the pH sensitivity of gap junctions: a model for synergistic interactions among connexins. AB - Gap junctions are formed by oligomerization of a protein called connexin. Most cells express more than one connexin isotype. Atrial myocytes, for example, coexpress connexin (Cx) 40 and Cx43. The consequence of connexin coexpression on the regulation of gap junctions is not well understood. In the present study, we show that cells coexpressing Cx40 and Cx43 are more susceptible to acidification induced uncoupling than those cells expressing only one connexin isotype. Xenopus oocytes were injected with mRNA for Cx40, Cx43, or a combination of both. Intracellular pH and junctional conductance were simultaneously measured while cells were progressively acidified by superfusion with a bicarbonate-buffered solution gassed with increasing concentrations of carbon dioxide. The data show that the pKa (ie, the pH at which junctional conductance decreased to 50% from maximum) shifted from approximately 6.7 when cells expressed only Cx40 or only Cx43 to approximately 7.0 when one of the oocytes was coexpressing both connexins. Truncation of the carboxyl terminal domains of the connexins caused the loss of pH sensitivity even after coexpression. The data are interpreted on the basis of previous studies from our laboratory that demonstrated heterodomain interactions in the regulation of Cx40 and Cx43 gap junctions. The possible implications of these findings on the regulation of native gap junctions that express both connexins remain to be determined. PMID- 10827143 TI - Hepatitis C, interferon alfa, and depression. PMID- 10827144 TI - Mimicry peptides of human PDC-E2 163-176 peptide, the immunodominant T-cell epitope of primary biliary cirrhosis. AB - The human PDC-E2 163-176 peptide (GDLLAEIETDKATI) is an immunodominant autoreactive T-cell epitope in patients with primary biliary cirrhosis (PBC), restricted by HLA DRB4*0101. We have previously reported that the ExDK sequence is essential for recognition of this epitope and identified 1 mimicry peptide, Escherichia coli PDC-E2 peptide (EQSLITVEGDKASM), which can activate human PDC-E2 163-176 peptide-reactive T-cell clones. In the present study, to further investigate mimicry peptides possibly involved in PBC, we generated 13 different T-cell clones reactive to the human PDC-E2 163-176 peptide following repeated in vitro stimulation of peripheral T lymphocytes with the human PDC-E2 163-176 peptide (native peptide) and tested for the reactivity of these T-cell clones to 30 different mimicry peptides derived from various self- and nonself proteins that have an ExDK-sequence. We found 7 mimicry peptides derived from microbial proteins that can activate at least 1 of these T-cell clones; 7 of 7 T-cell clones from patients with PBC and 2 of 6 T-cell clones from healthy subjects were activated by at least 1 to 6 different mimicry peptides. Two of 6 T-cell clones from healthy subjects were activated by specific mimicry peptides more strongly than by the native peptide, and 2 of 6 T-cell clones from healthy subjects were not activated by any mimicry peptides tested. Thus, the pattern and degree of activation by mimicry peptides differed in each T-cell clone, indicating the presence of a diverse spectrum of autoreactive T cells that are reactive to a single minimal epitope of the human PDC-E2 163-176 peptide. PMID- 10827145 TI - Decreased carnitine biosynthesis in rats with secondary biliary cirrhosis. AB - Carnitine biosynthesis was investigated in rats with secondary biliary cirrhosis induced by bile duct ligation (BDL) for 4 weeks (n = 5) and in pair-fed, sham operated control rats (n = 4). Control rats were pair-fed to BDL rats, and all rats were fed an artificial diet with negligible contents of carnitine, butyrobetaine, or trimethyllysine. Biosynthesis of carnitine and its precursors was determined by measuring their excretion in urine and accumulation in the body of the animals. Four weeks after BDL, total carnitine content was increased by 33% in livers from BDL rats when compared with control rats, but was unchanged in skeletal muscle and whole carcass. The plasma total carnitine concentration averaged 29.0 +/- 4.1 vs. 46.4 +/- 7.3 micromol/L in BDL rats and control rats, respectively. Urinary total carnitine excretion was reduced by 56% in BDL rats as compared with control rats. Carnitine biosynthesis was significantly decreased in BDL rats (0.45 +/- 0.19 vs. 0.93 +/- 0.08 micromol/100 g body weight/d in BDL and control rats, respectively). The tissue content of free and protein-linked trimethyllysine, a carnitine precursor, and trimethyllysine plasma concentrations were not different between BDL and control rats. However, urinary trimethyllysine excretion was increased 5-fold in BDL rats and approximated glomerular filtration. In contrast, urinary excretion of butyrobetaine, the direct carnitine precursor, was decreased by 40% in BDL rats as compared with control rats. Trimethyllysine biosynthesis was not different, but butyrobetaine biosynthesis was decreased by 51% in BDL as compared with control rats. In conclusion, carnitine biosynthesis is decreased in BDL rats as a result of a defect in the conversion of trimethyllysine to butyrobetaine. PMID- 10827146 TI - Increased natriuretic efficiency of furosemide in rats with carbon tetrachloride induced cirrhosis. AB - The authors examined the natriuretic efficiency of furosemide in rats with cirrhosis induced by carbon tetrachloride (CCl(4)). Rats were treated for 17 weeks with intraperitoneal injections of CCl(4) in groundnut oil twice a week throughout the study. Control rats were treated with vehicle (groundnut oil). Studies in metabolic cages showed that sodium retention was present from week 14. Renal clearance experiments were performed in chronically, instrumented conscious rats at the end of week 14 and at the termination of the study (end week 16) when ascites and hyponatremia were present. After 14 weeks, cirrhotic rats had sodium retention along with increased renal plasma flow, normal GFR, normal renal lithium handling, and a significantly increased diuretic (+41% vs. control) and natriuretic (+56% vs. control) response to a test dose furosemide (7.5 mg/kg b.w., intravenously). The natriuretic efficiency of furosemide, i.e., the natriuresis expressed relative to the furosemide excretion rate (triangle upU(Na)V/U(FUR)V) was increased by 51% versus control. After 17 weeks, ascites and hyponatremia had developed, and significant decreases in renal plasma flow ( 33%), GFR (-30%), and fractional lithium excretion (-44%) were observed. At this stage urinary recovery of furosemide was significantly decreased and the diuretic (-27% vs. Control) and natriuretic (-38% vs. control) responses to furosemide were significantly impaired. However, the increased natriuretic efficiency of furosemide was still present (+34% vs. control). Together these results suggest that increased sodium reabsorption in the thick ascending limb of Henle's loop is involved in the renal sodium retention in cirrhosis in rats that eventually results in decompensation with the formation of ascites. PMID- 10827147 TI - Autoimmune cholangitis within the spectrum of autoimmune liver disease. AB - Autoimmune cholangitis is an idiopathic disorder with mixed hepatocellular and cholestatic findings. Our goal was to characterize the disease prospectively by application of uniform diagnostic criteria. Twenty patients were identified and compared with 242 patients with conventional forms of autoimmune liver disease. Patients with autoimmune cholangitis were distinguished from type 1 autoimmune hepatitis (AIH) by lower serum levels of aspartate transaminase (AST), gamma globulin, and immunoglobulin G; higher serum levels of alkaline phosphatase; and lower frequencies of autoantibodies. They were distinguished from primary biliary cirrhosis (PBC) by higher serum levels of AST and bilirubin, lower serum concentrations of immunoglobulin M, and greater occurrence of autoantibodies. Their female predominance, lower serum alkaline phosphatase levels, higher frequency of autoantibodies, and absence of inflammatory bowel disease differentiated them from primary sclerosing cholangitis (PSC). Laboratory findings ranged widely and did not characterize individual patients. HLA risk factors were similar to those of type 1 AIH and PBC, and different from those of PSC. Treatment responses to corticosteroids or ursodeoxycholic acid were poor. Composite histological patterns resembled mainly PBC or PSC. We conclude that autoimmune cholangitis diagnosed by prospective analysis cannot be assimilated into a single, conventional, diagnostic category. It may represent variant forms of diverse conditions, a transition stage, or a separate entity with varying manifestations. PMID- 10827148 TI - Isosorbide mononitrate and propranolol compared with propranolol alone for the prevention of variceal rebleeding. AB - The aim of this study was to test the effectiveness of isosorbide-5-mononitrate (IM) as an adjunct to propranolol (PR) in the prevention of variceal rebleeding. Ninety-five cirrhotic patients with variceal bleeding were randomly assigned to treatment with PR + IM (46 patients) or PR alone (49 patients). Eighteen patients in the PR + IM group and 28 in the PR group had rebleeding during the 2 years after randomization. The actuarial probability of rebleeding 2 years after randomization was lower in the PR + IM group (40.4% vs. 57.4%) but the difference was not significant (P =. 09). However, the decrease in the risk of rebleeding reached statistical significance after stratification according to age, i.e. less than 50 versus >/=50 years old, (P =.03) or by adding an additional year of follow-up (P =.05). No significant difference was found in rebleeding index and survival. The multivariate Cox analysis indicated first, that both treatment (P =.03) and age (P =. 001) were factors predictive of rebleeding and second, that PR + MI reduced the risk of rebleeding by half (relative risk: 0.51, 95% confidence interval: 0.28-0.95). Seven patients in the PR + MI group and 1 patient in the PR group had to discontinue one of the drugs because of adverse events (P =.03). These results suggest that the addition of IM improves the efficacy of PR alone in the prevention of variceal rebleeding in cirrhotic patients. However no beneficial effects were observed on other parameters reflecting the efficacy of treatment. PMID- 10827149 TI - Quantitative evaluation of genomic instability as a possible predictor for development of hepatocellular carcinoma: comparison of loss of heterozygosity and replication error. AB - Both loss of heterozygosity (LOH) and replication error (RER) are considered to be phenotypes of genomic instability. To unveil the role of the genomic instability in hepatocarcinogenesis, frequencies of LOH and RER were simultaneously determined in 15 hepatocellular carcinomas (HCCs), surrounding nontumorous liver tissues (SL), and 13 liver tissues with chronic viral hepatitis void of cancer (NC) by referencing peripheral blood leukocytes (PBLs) from the corresponding donor using 18 microsatellite markers spread throughout the genome. LOH was significantly frequent in HCC compared with that in SL or NC (P =.005, P =.0003, respectively) and observed preferentially at particular microsatellite loci, D1S204, D2S123, D8S1106, D9S266, D16S748, and D19S601. Although the higher prevalence of RER was also significant in HCC compared with that in NC (P =.03), in most cases the errors were detected at very low frequencies and random loci. Both LOH and RER tended to appear more prevalently in SL than in NC. The occurrence rate of LOH was higher in the tissues associated with hepatitis B virus (HBV) than with hepatitis C virus (HCV) infection especially in HCC (P =.03). When referencing SL instead of PBLs, the prevalence of LOH and RER in HCC significantly decreased (P =.02 and P =.03, respectively). These results suggest that LOH is closely associated with multistep hepatocarcinogenesis especially under HBV infection, but RER is imperceptibly associated. The quantitative evaluation of the frequency of LOH by referencing PBLs may be a useful predictor for HCC development in chronic liver diseases. PMID- 10827150 TI - Differential expression of lymphoid and myeloid markers on differentiating hematopoietic stem cells in normal and tumor-bearing adult human liver. AB - The presence and phenotype of lineage-committed hematopoietic progenitors in the normal adult human liver (AHL) were investigated and compared with the profiles of differentiating hematopoietic precursor populations detected in liver bearing metastases of colonic origin. Levels of hematopoietic stem cells (HSCs) (CD34(+)CD45(+)) detected in hepatic mononuclear cell (HMNC) populations were increased 6-fold when compared with matched peripheral blood samples. In normal liver, less than 5% of HSCs expressed the myeloid-associated antigen, CD33, whereas considerable proportions expressed lymphoid-associated markers (T cell, 33.39%; B cell, 17.39%; and natural killer [NK] cell, 37.17%). Significant increases were observed in the relative proportions of hepatic HSCs coexpressing CD33 (20.53%; P =.001), and the T-cell marker (CD7, 58. 13%; P =.02) in tumor bearing liver compared with normal liver. HSCs with B-cell progenitor phenotype (CD19(+)) were significantly decreased in tumor-bearing liver (0.06%; P =.02). Despite these differences, the activation status of hematopoiesis, as measured by the coexpression of the differentiation and activation markers, CD38 and CD45RA, did not differ significantly between normal and tumor-bearing liver. These results indicate that the normal AHL harbors lineage-committed hematopoietic progenitors, and the vast majority of these progenitors express lymphoid associated antigens with changes occurring in both the myeloid and lymphoid compartments of the hepatic hematopoietic pathway on tumor challenge. While tumor bearing livers are enriched for intrahepatic myeloid precursors and T-cell progenitor cells, further studies are required to establish the origin and in situ development potential of hepatic HSCs in the adult human and their role in tumor immunity. PMID- 10827151 TI - Unique epithelial cell production of hepatocyte growth factor/scatter factor by putative precancerous intestinal metaplasias and associated "intestinal-type" biliary cancer chemically induced in rat liver. AB - Recently, we observed that Met, the receptor for hepatocyte growth factor/scatter factor (HGF/SF), is overexpressed in epithelial cells of both early-appearing intestinal metaplastic glands in precancerous hepatic cholangiofibrotic tissue and neoplastic glands in later developed intestinal-type of cholangiocarcinoma originated from the furan rat model of cholangiocarcinogenesis when compared with normal and hyperplastic intrahepatic biliary epithelia. We now show that HGF/SF is also aberrantly expressed in a manner closely paralleling that of its receptor in the neoplastic epithelial cells of furan-induced rat cholangiocarcinomas and in a majority of metaplastic epithelial cells within earlier formed precancerous hepatic cholangiofibrotic tissue. Using in situ hybridization and reverse transcription-polymerase chain reaction (RT-PCR), we further showed specific expression of HGF/SF messenger RNA (mRNA) in a novel rat cholangiocarcinoma epithelial cell line overexpressing Met. This cholangiocarcinoma cell line, termed C611B, was established from tumorigenic cells isolated from a furan induced transplantable tumor. Moreover, we detected by in situ hybridization strong expression of HGF/SF mRNA transcripts in the cancerous epithelial glands of cholangiocarcinoma developed in recipient rats after in vivo cell transplantation of C611B cells. In contrast, mRNA transcripts and protein immunoreactivity for this cytokine were not detected in hepatocytes and biliary epithelial cells in adult normal rat liver nor in rat hyperplastic intrahepatic biliary epithelium. Our results clearly show that HGF/SF becomes aberrantly expressed in cholangiocarcinoma epithelium and in putative precancerous intestinal metaplastic epithelium induced in the liver of furan-treated rats. PMID- 10827152 TI - Cold liver ischemia-reperfusion injury critically depends on liver T cells and is improved by donor pretreatment with interleukin 10 in mice. AB - Kupffer cells are thought to mediate most of the deleterious effects of liver ischemia-reperfusion injury. The role of liver T cells and the impact of resident cell deactivation by interleukin 10 (IL-10) have never been addressed. Using a model of ex vivo liver cold ischemia and reperfusion, we assessed liver injury, tumor necrosis factor (TNF) and interferon gamma (IFN-gamma) release from livers of balb/c mice, nude mice, nude mice reconstituted with T cells, and gadolinium balb/c pretreated mice. The anti-inflammatory cytokine IL-10 was then used to define the best strategy of administration potentially able to modulate ischemia reperfusion injury. For this purpose IL-10 was administered to the donor before liver harvesting, in the preservation medium during cold ischemia or during reperfusion. TNF and IFN-gamma were released time dependently and paralleled liver injury after reperfusion of cold preserved livers. Reperfused livers from nude or gadolinium pretreated mice disclosed a dramatic decrease in TNF and IFN gamma release. Tissue injury was reduced by 51% in the absence of T cells and by 88% when Kupffer cells were deactivated. This effect was reverted by T-cell transfer to nude mice. Only donor pretreatment with IL-10 or IL-10 infusion during reperfusion led to a significant decrease in liver injury, TNF, and IFN gamma release (-66% or -41%, -95% or -94%, and -70% or -70%, respectively). In conclusion, liver resident T cells are critically involved in cold ischemia reperfusion injury and pretreatment of the donor with IL-10 decreases liver injury and the release of T-cell- and macrophage-dependent cytokines. PMID- 10827153 TI - Interferon tau-induced hepatocyte apoptosis in sheep. AB - Clinical applications of Type I interferon (IFN) are limited by adverse side effects mediated largely by unknown mechanisms. This study examined the mechanisms of acute hepatic injury in lambs treated with systemic administration of IFN-tau, a Type I IFN. Liver tissues were collected at 24, 48, or 96 hours after treatment with either IFN-tau or saline. Histopathology revealed acute hepatopathy including cellular swelling, cytoplasmic aggregates, and apoptosis in all IFN-tau-treated lambs, which were accompanied by elevation of aspartate transaminase (AST) (P <.01). The number of apoptotic hepatocytes in IFN-tau treated lambs was higher than for control lambs (P <.001). Immunohistochemistry for proliferating cell nuclear antigen (PCNA) revealed that IFN-tau induced hepatocyte growth arrest at the G0/G1 phase of the cell cycle and that the majority of hepatocytes in S or G2 phase were eliminated by apoptosis. We investigated expression of bax-alpha and bcl-2, acting as pro- and antiapoptotic molecules, in IFN-tau-induced apoptosis. Northern blot analysis revealed increased expression of bax-alpha messenger RNA (mRNA) in IFN-tau-treated lambs (P <.01) compared with control lambs, consistent with the expression pattern for bax-alpha protein. However, there was no detectable difference in expression of bcl-2 proteins between control and IFN-tau-treated lambs. The levels of bax-alpha associated with the mitochondria also increased during IFN-tau treatment. Bax alpha immunostaining showed scattered immunoreactive hepatocytes with morphological hallmarks of apoptosis. These results suggest that IFN-tau induces growth arrest as well as apoptosis by regulating bax-alpha expression. These pathological effects of IFN-tau on sheep liver indicate potential mechanisms of Type 1 IFN-induced hepatotoxicity in animals and humans. PMID- 10827154 TI - Cholestasis with altered structure and function of hepatocyte tight junction and decreased expression of canalicular multispecific organic anion transporter in a rat model of colitis. AB - Cholestasis is frequently associated with inflammatory bowel disease. Because some cholestasis is resulted from altered hepatocyte tight junctions (TJs) or the canalicular multispecific organic anion transporter, we have investigated the following topics in a rat model of inflammatory bowel disease: (1) alterations in hepatocyte TJs and in the canalicular multispecific organic anion transporter, (2) etiologic factors for cholestasis, and (3) effects of antibiotics on cholestasis. Rats with trinitrobenzene sulfonic acid-induced colitis were studied 24 hours after treatment. Hepatocyte TJs and the canalicular multispecific organic anion transporter were evaluated by immunostaining for TJ-associated proteins, 7H6 and ZO-1, and multidrug resistance protein 2 (mrp2). To investigate etiologic factors causing cholestasis, portal endotoxin and proinflammatory cytokines were examined. The effects of polymyxin B, penicillin G, or metronidazole on immunostaining for 7H6, ZO-1, mrp2, and cholestasis were investigated. (1) Immunostaining for 7H6 and ZO-1 colocalized outlining the bile canaliculi and immunostaining for mrp2 localized on the canalicular membrane in controls. Treatment with trinitrobenzene sulfonic acid induced significant cholestasis and caused translocation of immunostaining for 7H6, but not that for ZO-1, to the cytoplasm and diminished immunostaining for mrp2 on the canaliculus membrane. (2) The levels of portal endotoxin, but not proinflammatory cytokines, was increased. (3) Polymyxin B, but not the other antibiotics, prevented alterations in immunostaining for both 7H6 and mrp2, and cholestasis. We described that both hepatocyte TJs and the canalicular multispecific organic anion transporter were altered and that gut-derived endotoxin levels in the portal blood were increased in this rat colitis model. PMID- 10827155 TI - Cloning and functional characterization of the bile acid-sensitive methotrexate carrier from rat liver cells. AB - We have cloned two complementary DNAs (cDNAs), RL-Mtx-1 and RL-Mtx-2, corresponding to the bile acid- sensitive methotrexate carrier from rat liver by direct full-length rapid amplification of cDNA ends polymerase chain reaction (RACE-PCR) using degenerated primers that were deduced from published sequences of tumor cell methotrexate transporters. When expressed in Xenopus laevis oocytes and cosM6 cells, both clones mediate methotrexate and bumetanide transport. RL Mtx-1 consists of 2,445 bp with an open reading frame of 1,536 bp. The corresponding protein with 512 amino acids has a molecular weight of 58 kd. RL Mtx-2 (2,654 bp) differs by an additional insert of 203 bp. This insert is located in frame at position 1,196 of the RL-Mtx-1 and contains the typical splice junction sites at the 5' and 3' end, indicating that the RL-Mtx-2 messenger RNA (mRNA) is generated by alternative splicing. The insert contains a stop codon that shortens the RL-Mtx-2 protein to 330 amino acids (38 kd). Both cDNAs contain the binding site sequence for the dioxin/nuclear translocator responsive element (Ah/Arnt-receptor) in conjunction with a barbiturate recognition sequence (Barbie box). Preliminary results show that the Barbie box acts as a negative regulatory element. The two liver cDNA clones show homologies to the published sequences of folate and the reduced folate carriers, but no homology is found to the transport systems for organic anions like the Ntcp1, oatp1, OAT-K1, and OAT1. Expression of the mRNA for the methotrexate carrier is found in liver, kidney, heart, brain, spleen, lung, and skeletal muscle, but not in the testis as revealed by Northern blot analysis. The highest abundance of the mRNA is found in the kidney. PMID- 10827156 TI - Bile acid synthesis in cultured human hepatocytes: support for an alternative biosynthetic pathway to cholic acid. AB - The biosynthesis of bile acids by primary cultures of normal human hepatocytes has been investigated. A general and sensitive method for the isolation and analysis of sterols and bile acids was used, based on anion exchange chromatography and gas chromatography-mass spectrometry (GC/MS). Following incubation for 5 days, 8 oxysterols and 8 C(27)- or C(24)-bile acids were identified in media and cells. Cholic and chenodeoxycholic acids conjugated with glycine or taurine were by far the major steroids found, accounting for 70% and 24% of the total, respectively, being consistent with bile acid synthesis in human liver. Small amounts of sulfated 3beta-hydroxy-5-cholenoic acid and 3beta,7alpha-dihydroxy-5beta-cholanoic acid were also detected. Nine steroids were potential bile acid precursors (2% of total), the major precursors being 7alpha, 12alpha-dihydroxy-3-oxo-4-cholenoic acid and its 5beta-reduced form. These 2 and 5 other intermediates formed a complete metabolic sequence from cholesterol to cholic acid (CA). This starts with 7alpha-hydroxylation of cholesterol, followed by oxidation to 7alpha-hydroxy-4-cholesten-3-one and 12alpha-hydroxylation. Notably, 27-hydroxylation of the product 7alpha, 12alpha dihydroxy-4-cholesten-3-one and further oxidation and cleavage of the side chain precede A-ring reduction. A-Ring reduction may also occur before side-chain cleavage, but after 27-hydroxylation, yielding 3alpha,7alpha, 12alpha-trihydroxy 5beta-cholestanoic acid as an intermediate. The amounts of the intermediates increased in parallel to those of CA during 4 days of incubation. Suppressing 27 hydroxylation with cyclosporin A (CsA) resulted in a 10-fold accumulation of 7alpha, 12alpha-dihydroxy-4-cholesten-3-one and a decrease of the production of CA and its acidic precursors. These results suggest that the observed intermediates reflect an alternative biosynthetic pathway to CA, which may be quantitatively significant in the cells. PMID- 10827157 TI - Expression of aquaporin-4 water channels in rat cholangiocytes. AB - We recently reported that secretin induces the exocytic insertion of functional aquaporin-1 water channels (AQP1) into the apical membrane of cholangiocytes and proposed that this was a key process in ductal bile secretion. Because AQP1 is present on the basolateral cholangiocyte membrane in low amounts, we hypothesized that another AQP must be expressed at this domain to facilitate transbasolateral water movement. Thus, we investigated the expression, subcellular localization, possible regulation by secretin, and functional activity of AQP4, a mercury insensitive water channel expressed in other fluid transporting epithelia. Using reverse transcription-polymerase chain reaction (RT-PCR) on RNA prepared from purified rat cholangiocytes, we amplified a product of 311 bp that was 100% homologous to the reported AQP4 sequence. RNase protection assay confirmed the presence of an appropriate size transcript for AQP4 in cholangiocytes. Immunoblotting detected a band of approximately 31 kd corresponding to AQP4 in basolateral but not apical membranes of cholangiocytes. Secretin did not alter the amount of plasma membrane AQP4 but, as expected, induced AQP1 redistribution from intracellular to apical plasma membranes. Functional studies showed that AQP4 accounts for about 15% of total cholangiocyte membrane water permeability. Our results indicate that: (1) cholangiocytes express AQP4 messenger RNA (mRNA) and protein and (2) in contrast to AQP1, which is targeted to the apical cholangiocyte membrane by secretin, AQP4 is constitutively expressed on the basolateral cholangiocyte membrane and is secretin unresponsive. The data suggest that AQP4 facilitates the basolateral transport of water in cholangiocytes, a process that could be relevant to ductal bile formation. PMID- 10827158 TI - Clearance of the original hepatitis B virus YMDD-motif mutants with emergence of distinct lamivudine-resistant mutants during prolonged lamivudine therapy. AB - Tyrosine-methionine-aspartate-aspartate (YMDD)-motif mutants may emerge and elicit immune clearance during prolonged lamivudine treatment. The aim of this study was to investigate the virological events following development of the original mutants. Twenty-three patients who developed YMDD-motif mutants during the Asian lamivudine trial were included. Serial serum samples from these patients were subjected to sequence analysis to identify new mutants. Site directed mutagenesis experiments were performed to investigate whether the new mutations were responsible for lamivudine resistance. Of the 23 patients included, 13 harbored either one or a mixture of the two common YMDD-motif mutants (methionine 552-to-isoleucine [M552I] and leucine 528-to methionine/methionine 552-to-valine [L528M/M552V]) throughout the course, whereas in the remaining 10 patients, distinct mutants became dominant over the original mutants to cause continuing chronic hepatitis. Of them, 3 developed an alanine 529-to-threonine (A529T) mutant, 6 developed a leucine 528-to methionine/methionine 552-to-isoleucine (L528M/M552I) mutant, and 1 developed these two mutants sequentially. Site-directed mutagenesis experiments confirmed that the aforementioned mutations were responsible for the resistance to lamivudine in vitro. The nucleotide substitution in the A529T mutant concomitantly generated a stop codon at the surface gene, leading to impaired secretion of HBsAg. Strikingly, the replication of this mutant was lamivudine dependent. These results suggested that distinct lamivudine-resistant mutants could emerge and replace the original YMDD-motif mutants as the cause of continuing chronic hepatitis during prolonged lamivudine therapy. PMID- 10827159 TI - Priming of hepatitis C virus-specific cytotoxic T lymphocytes in mice following portal vein injection of a liver-specific plasmid DNA. AB - The immunology of hepatitis C virus (HCV) infection should be studied in the context of HCV antigen expression in the liver, because HCV primarily infects this organ. Indeed, the nature, function, and fate of T cells primed after antigen expression in the liver might differ from those primed when antigens are expressed systemically or in other organs, because the nature of the antigen presenting cells (APCs) involved may be different. In addition, the normal liver contains a resident population of lymphocytes that differ from those present at other sites. Thus, we investigated whether HCV-specific CD8(+) cytotoxic T cells (CTLs) could be elicited following portal vein (PV) injection of plasmid DNA in mice whose hepatic veins were transiently occluded. We show that PV injection of mice with "naked" DNA expressing the HCV-NS5a protein, under the control of a liver-specific enhancer/promoter, resulted in NS5a expression in the liver and the priming of HCV-specific CTLs. These results suggested that such a model might be relevant to the study of HCV-specific immune responses primed during natural infection. PMID- 10827160 TI - Viral clearance in hepatitis C (1b) infection: relationship with human leukocyte antigen class II in a homogeneous population. AB - The aim of this study was to investigate the possibility of a significant relationship between human leukocyte antigen (HLA) class II and the clearance of hepatitis C virus (HCV). The study group consisted of 156 Irish women who iatrogenically received HCV 1b-contaminated Anti-D immunoglobulin between May 1977 and November 1978. Thus, the study population was homogeneous in terms of gender, source of infection, and ethnicity. On Screening in 1994, all individuals were anti-HCV antibody positive by recombinant immunoblot assay, while 46% (n = 72) of the group were HCV-positive by reverse transcriptase-polymerase chain reaction (RT-PCR). HLA DRB1 and DQB1 status was molecularly defined by high resolution reverse line probe hybridization methodology. Clearance of HCV 1b was found to be associated with DRB1*01. However, this association was lost after Bonferroni correction for multiple comparisons. Extended haplotype analysis between specific DRB1 and DQB1 allelic combinations identified a significant reduction in the frequency of DQB1*0501 in the presence of DRB1*0701 in the persistently infected individuals in the study group (P <.05). No associations with either viral clearance or persistence were found at the DQB1 locus. Our results suggest that HLA DRB1*01 appears to contribute to the spontaneous resolution of a primary HCV infection in the Irish population. The presence of DRB1*0701 in the absence of DQB1*0501 possibly reflects an influence of this allele in persistence of HCV infection. Defined and homogeneous patient populations offer the best opportunity to illuminate previously disguised immunogenetic factors important in the clearance of HCV 1b. PMID- 10827161 TI - Combination of ribavirin and interferon-alfa surpasses high doses of interferon alfa alone in patients with genotype-1b-related chronic hepatitis C. AB - The purpose of this study was to compare interferon-alfa alone (12-month course with high initial doses) with a combination of interferon-alfa and ribavirin in patients infected with genotype 1b. Three hundred and seven patients were randomized into 3 groups to receive 6 mega units (MU) of interferon-alfa-2b subcutaneously 3 times weekly for 6 months followed by 3 MU for 6 months (n = 95, group A); 10 MU for 3 months followed by 6 MU for 3 months, followed by 3 MU for 6 months (n = 83, group B); or the group-A schedule in combination with ribavirin (n = 129, group C) for 4 (n = 46), 6 (n = 44), or 12 months (n = 39). Negative polymerase chain reaction (PCR) was more frequent in group C than in groups A or B after 3 months of treatment (P <.006), at the end of treatment (P =.017), and at the end of follow-up (32.8%, 16.9%, and 14.1%, respectively, P <.003). A complete response (negative PCR and normal alanine transaminase) was higher in group C than in the other groups and when comparing 12- to 4- and 6-month combination therapy at the end of treatment (P =.05) and of follow-up (45.2% vs. 25.4%, respectively, P =.05). The greater efficacy of the combination was related to the higher rate of primary virological response and also to a decrease in the percentage of breakthrough and of relapse. In 1b-infected patients, the combination of high doses of interferon-alfa (6 MU) and ribavirin for 12 months appears to be the best therapy, with a high rate of sustained response. PMID- 10827162 TI - Influence of immunogenetic background on the outcome of recurrent hepatitis C after liver transplantation. AB - In immunocompetent patients, specific human leukocyte antigen (HLA) class II alleles have been associated with the severity of hepatitis C virus (HCV)-related disease, in particular, HLA-DRB1*11 has been found to exert a protective effect. The authors have analyzed the role of HLA class I and II alleles in determining the frequency, timing, and progression of histologically proven recurrent hepatitis C in 89 patients who underwent a liver transplant for HCV-related cirrhosis. In addition, the influence of HLA mismatch between donor and recipient, HCV genotype, and use of steroid pulses was also evaluated. Median patient follow up was 35 months (range 4-119). HLA-DRB1 typing was performed by genomic analysis in all cases. Liver biopsies were obtained routinely and at least at yearly intervals. Histologically proven recurrent hepatitis was observed in 46 patients (52%), 10 patients progressing to stage 5-6 fibrosis in most cases within 2 years after transplant. By univariate analysis, 3 variables, HLA-B14, HLA-DRB1*04, and HLA-DRB1 donor/recipient mismatch, showed a significant effect on time to recurrent hepatitis C disease. These parameters were included in a multivariate regression model along with HCV genotype, treatment with steroid pulses and DRB1*11. HLA-B14, HLA-DRB1*04, and HLA-DRB1 donor/recipient mismatch were confirmed to provide a significant and independent contribution to the risk of hepatitic disease recurrence. As for the severity of the disease, none of the 10 patients with stage 5-6 hepatitis carried the HLA-DRB1*11 allele, in line with what was observed in nontransplant subjects. Our results suggest that in posttransplant recurrent hepatitis C, immunogenetic factors are relevant in determining HCV infection outcome. PMID- 10827163 TI - Hepatitis C virus core protein induces apoptosis and impairs cell-cycle regulation in stably transformed Chinese hamster ovary cells. AB - Hepatitis C virus (HCV) infection is associated with the development of hepatocellular carcinoma. Several lines of evidence suggest that the core protein of HCV may play a role in the development of this cancer. The authors examined regulation of the cell cycle in stable cell lines derived from Chinese hamster ovary (CHO-K1) cells that constitutively expressed one or more of the structural proteins of HCV. In media containing low concentrations of serum (serum starvation), cell lines expressing the core protein showed a significantly lower population of viable cells than noncore-expressing cells. The low viability of the core-expressing cells was a result of the increased population of cells undergoing apoptosis. Interestingly, the cell cycle analysis revealed that the arresting function at G(0) was impaired, and the cell cycle was accelerated in core-expressing cell lines even under serum starvation. Thus, the HCV core protein sensitizes the apoptosis to serum starvation, although it promotes the cell cycle in CHO-K1 cells. To explain these findings, the authors examined the expression of revival apoptosis and cell-cycle-related genes. Expression of the c myc genes was significantly induced in core-expressing cells in response to serum starvation. Other apoptosis-inducing genes downstream of c-myc, p53, p21WAF1/CIP1 and Bax were significantly highly induced, although there was no induction of Bcl 2, which prevents apoptosis in core-expressing cells. Thus, the HCV core protein induced apoptosis and impaired the regulation of the cell cycle by activating c myc expression, whereas the p53 and Bax pathways play a role in the induction of apoptosis. PMID- 10827164 TI - Mutations within the E2 and NS5A protein in patients infected with hepatitis C virus type 3a and correlation with treatment response. AB - Defined regions of hepatitis C virus (HCV) envelope 2 (E2), PePHD, and nonstructural 5A (NS5A) protein (PKR-binding domain) have been shown to interact with interferon alfa (IFN-alpha)-inducible double-stranded RNA-activated protein kinase (PKR) in vitro, suggesting a possible mechanism of HCV to evade antiviral effects of IFN-alpha. The clinical correlation between amino acid mutations within the E2 PePHD or the NS5A PKR-binding domain and response to antiviral treatment in HCV-3a-infected patients is unknown. Thirty-three patients infected with HCV-3a isolates were treated with IFN-alpha with or without ribavirin. The carboxyterminal half of E2 and of the NS5A gene were sequenced. Sixteen patients achieved a sustained virological response (SR), 6 patients an end-of-treatment response with relapse thereafter (ETR), and 11 patients were nonresponders (NR). Within the PePHD of the E2 protein 0.5 (range, 0-2) mutations were observed in SR patients, whereas the number of mutations in ETR or NR patients was 0.2 (0-1). Quasispecies analyses showed almost no heterogeneity. The mean number of mutations within the PKR-binding domain of the NS5A protein was 1.6 (range, 0-4) in SR patients, 1 (0-2) in ETR patients, and 1.6 (0-3) in NR patients. Patients with higher numbers of mutations within the E2 or NS5A region showed a trend towards lower pretreatment viremia. Phylogenetic and conformational analyses of E2 or NS5A sequences allowed no differentiation between sensitive and resistant isolates. However, mutations within the E2 PePHD in SR patients were frequent, and hydrophobic mutations within the hydrophilic area of PePHD at codon 668 and 669 were exclusively observed in sustained virological responders. PMID- 10827165 TI - Association between hepatitis B virus infection and HLA-DR type in Korea. AB - Although the mechanism of susceptibility to chronic persistent hepatitis B virus (HBV) infection is not well clarified, immunogenetic factors of the host may have a role. Recently, a strong association between HLA-DR13 and the self-limited course of HBV infection has been reported. To determine whether the elimination of HBV is related to a particular HLA allele, we studied the HBV markers and HLA DR phenotypes of 1,272 Koreans who had visited Yonsei University Medical Center for renal transplantation. They included 330 renal transplant donors. Subjects were categorized into 3 different groups: the "Unexposed Group" (UE; n = 946) with negative HBV markers, the "Chronic Carrier Group" (CC; n = 83), who were hepatitis B surface antigen (HBsAg)-positive, and the "Spontaneously Cleared Group" (SC; n = 243), who were HBsAg-negative with antibodies to HBsAg (anti-HBs) and hepatitis B core antigen (anti-HBc). HLA-DR4 was the most common type in all groups. HLA-DR6 was significantly more frequent in 69 of 243 subjects with SC (28. 4%) than in 8 of 83 subjects with CC (9.6%) (P <.001; relative risk [RR] = 3.72). HLA-DR9 was significantly more frequent in CC than in SC (P <.001; RR = 0.33). HLA-DR13 showed a stronger association with the clearance of HBV than the other HLA-DR6 subgroup. The distribution of HLA-DR phenotypes was similar regardless of renal disease. Our data indicate that HLA-DR6, especially HLA-DR13, is one of the host factors, which influences the immune response to HBV, and may be associated with self-elimination of HBV in Koreans. PMID- 10827166 TI - Hepatic inflammation and immunity: a summary of a conference on the function of the immune system within the liver. PMID- 10827167 TI - Crystal structure of dTDP-4-keto-6-deoxy-D-hexulose 3,5-epimerase from Methanobacterium thermoautotrophicum complexed with dTDP. AB - Deoxythymidine diphosphate (dTDP)-4-keto-6-deoxy-d-hexulose 3, 5-epimerase (RmlC) is involved in the biosynthesis of dTDP-l-rhamnose, which is an essential component of the bacterial cell wall. The crystal structure of RmlC from Methanobacterium thermoautotrophicum was determined in the presence and absence of dTDP, a substrate analogue. RmlC is a homodimer comprising a central jelly roll motif, which extends in two directions into longer beta-sheets. Binding of dTDP is stabilized by ionic interactions to the phosphate group and by a combination of ionic and hydrophobic interactions with the base. The active site, which is located in the center of the jelly roll, is formed by residues that are conserved in all known RmlC sequence homologues. The conservation of the active site residues suggests that the mechanism of action is also conserved and that the RmlC structure may be useful in guiding the design of antibacterial drugs. PMID- 10827168 TI - Association of calcium/calmodulin-dependent kinase II with developmentally regulated splice variants of the postsynaptic density protein densin-180. AB - In a continuing search for proteins that target calcium/calmodulin-dependent protein kinase II (CaMKII) to postsynaptic density (PSD) substrates important in synaptic plasticity, we showed that the PSD protein densin-180 binds CaMKII. Four putative splice variants (A-D) of the cytosolic tail of densin-180 are shown to be differentially expressed during brain development. Densin-180 splicing affects CaMKII phosphorylation of specific serine residues. Variants A, B, and D, but not C, bind CaMKII stoichiometrically and with high affinity, mediated by a differentially spliced domain. Densin-180 differs from the previously identified CaMKII-binding protein NR2B in that binding does not strictly require CaMKII autophosphorylation. Binding of densin-180 and NR2B to CaMKII is noncompetitive, indicating different interaction sites on CaMKII. Expression of the membrane targeted CaMKII-binding domain of densin-180 confers membrane localization to coexpressed CaMKII without requiring calcium mobilization, suggesting that densin 180 plays a role in the constitutive association of CaMKII with PSDs. PMID- 10827169 TI - Nucleoside transporter proteins of Saccharomyces cerevisiae. Demonstration of a transporter (FUI1) with high uridine selectivity in plasma membranes and a transporter (FUN26) with broad nucleoside selectivity in intracellular membranes. AB - FUI1 and function unknown now 26 (FUN26) are proteins of uncertain function with sequence similarities to members of the uracil/allantoin permease and equilibrative nucleoside transporter families of transporter proteins, respectively. [(3)H]Uridine influx was eliminated by disruption of the gene encoding FUI1 (fui1) and restored by expression of FUI1 cDNA, whereas influx in transport-competent and fui1-negative yeast were unaffected, respectively, by disruption of the FUN26 gene or overexpression of FUN26 cDNA. FUI1 transported uridine with high affinity (K(m), 22 +/- 3 micrometer) and was unaffected or inhibited only partially by high concentrations (1 mm) of a variety of ribo- and deoxyribonucleosides or nucleobases. When FUN26 cDNA was expressed in oocytes of Xenopus laevis, inward fluxes of [(3)H]uridine, [(3)H]adenosine, and [(3)H]cytidine were stimulated, and uridine influx was independent of pH and not inhibited by dilazep, dipyridamole, or nitrobenzylmercaptopurine ribonucleoside. Fractionation of yeast membranes containing immunotagged recombinant FUN26 (shown to be functional in oocytes) demonstrated that the protein was primarily in intracellular membranes. These results indicated that FUI1 has high selectivity for uracil-containing ribonucleosides and imports uridine across cell-surface membranes, whereas FUN26 has broad nucleoside selectivity and most likely functions to transport nucleosides across intracellular membranes. PMID- 10827170 TI - Dual function of the propeptide of prouroguanylin in the folding of the mature peptide: disulfide-coupled folding and dimerization. AB - Guanylyl cyclase activating peptide II (GCAP-II), an endogenous ligand of guanylyl cyclase C, is produced via the processing of the precursor protein (prepro-GCAP-II). We have previously shown that the propeptide in pro-GCAP-II functions as an intramolecular chaperone in the proper folding of the mature peptide, GCAP-II (Hidaka, Y., Ohno, M., Hemmasi, B., Hill, O., Forssmann, W.-G., and Shimonishi, Y. (1998) Biochemistry 37, 8498-8507). Here, we report an essential region in pro-GCAP-II for the correct disulfide pairing of the mature peptide, GCAP-II. Five mutant proteins, in which amino acid residues were sequentially deleted from the N terminus, and three mutant proteins of pro-GCAP II, in which N-terminal 6, 11, or 17 amino acid residues were deleted, were overproduced using Escherichia coli or human kidney 293T cells, respectively. Detailed analysis of in vivo or in vitro folding of these mutant proteins revealed that one or two amino acid residues at the N terminus of pro-GCAP-II are critical, not only for the chaperone function in the folding but also for the net stabilization of pro-GCAP-II. In addition, size exclusion chromatography revealed that pro-GCAP-II exists as a dimer in solution. These data indicate that the propeptide has two roles in proper folding: the disulfide-coupled folding of the mature region and the dimerization of pro-GCAP-II. PMID- 10827171 TI - In vivo functional analysis of the human mitochondrial DNA polymerase POLG expressed in cultured human cells. AB - The human gene POLG encodes the catalytic subunit of mitochondrial DNA polymerase, but its precise roles in mtDNA metabolism in vivo have not hitherto been documented. By expressing POLG fusion proteins in cultured human cells, we show that the enzyme is targeted to mitochondria, where the Myc epitope-tagged POLG is catalytically active as a DNA polymerase. Long-term culture of cells expressing wild-type POLG-myc revealed no alterations in mitochondrial function. Expression of POLG-myc mutants created dominant phenotypes demonstrating important roles for the protein in mtDNA maintenance and integrity. The D198A amino acid replacement abolished detectable 3'-5' (proofreading) exonuclease activity and led to the accumulation of a significant load (1:1700) of mtDNA point mutations during 3 months of continuous culture. Further culture resulted in the selection of cells with an inactivated mutator polymerase, and a reduced mutation load in mtDNA. Transient expression of POLG-myc variants D890N or D1135A inhibited endogenous mitochondrial DNA polymerase activity and caused mtDNA depletion. Deletion of the POLG CAG repeat did not affect enzymatic properties, but modestly up-regulated expression. These findings demonstrate that POLG exonuclease and polymerase functions are essential for faithful mtDNA maintenance in vivo, and indicate the importance of key residues for these activities. PMID- 10827172 TI - Recognition of formamidopyrimidine by Escherichia coli and mammalian thymine glycol glycosylases. Distinctive paired base effects and biological and mechanistic implications. AB - The activity of prokaryotic and mammalian thymine glycol (Tg) glycosylases including Escherichia coli endonuclease III (Endo III) and endonuclease VIII (Endo VIII) and mouse Endo III homologue (mNth1) for formamidopyrimidine (Fapy) has been investigated using defined oligonucleotide substrates. 2, 6-Diamino-4 hydroxy-5-N-methylformamidopyrimidine, a methylated Fapy derived from guanine, was site specifically incorporated in the oligonucleotide. The substrates containing Fapy:N pairs (N = A, G, C, T) as well as a Tg:A pair, a physiological substrate of Endo III, Endo VIII, and mNth1, were treated by the enzymes and nicked products were quantified by gel electrophoresis. The activity of Endo III and Endo VIII for Fapy varied markedly depending on the paired base, being the highest with G (activity relative to Tg = 0. 55 (Endo III) and 0.41 (Endo VIII)) and the lowest with C (0.05 (Endo III) and 0.06 (Endo VIII)). In contrast, mNth1 recognized all Fapy pairs equally well and the activity was comparable to Tg. The results obtained in the nicking assay were further substantiated by the analysis of the Schiff base intermediate using NaBH(4) trapping assays. These results indicate that Escherichia coli and mammalian Tg glycosylases have a potential activity to recognize Fapy. However, as demonstrated for Fapy:C pairs, their distinctive activities implicate unequal participation in the repair of Fapy lesions in cells. PMID- 10827173 TI - Interactions of the low density lipoprotein receptor gene family with cytosolic adaptor and scaffold proteins suggest diverse biological functions in cellular communication and signal transduction. AB - The members of the low density lipoprotein (LDL) receptor gene family bind a broad spectrum of extracellular ligands. Traditionally, they had been regarded as mere cargo receptors that promote the endocytosis and lysosomal delivery of these ligands. However, recent genetic experiments in mice have revealed critical functions for two LDL receptor family members, the very low density lipoprotein receptor and the apoE receptor-2, in the transmission of extracellular signals and the activation of intracellular tyrosine kinases. This process regulates neuronal migration and is crucial for brain development. Signaling through these receptors requires the interaction of their cytoplasmic tails with the intracellular adaptor protein Disabled-1 (DAB1). Here, we identify an extended set of cytoplasmic proteins that might also participate in signal transmission by the LDL receptor gene family. Most of these novel proteins are adaptor or scaffold proteins that contain PID or PDZ domains and function in the regulation of mitogen-activated protein kinases, cell adhesion, vesicle trafficking, or neurotransmission. We show that binding of DAB1 interferes with receptor internalization suggesting a mechanism by which signaling through this class of receptors might be regulated. Taken together, these findings imply much broader physiological functions for the LDL receptor family than had previously been appreciated. They form the basis for the elucidation of the molecular pathways by which cells respond to the diversity of ligands that bind to these multifunctional receptors on the cell surface. PMID- 10827174 TI - The thrombospondin motif of aggrecanase-1 (ADAMTS-4) is critical for aggrecan substrate recognition and cleavage. AB - Aggrecanase-1 (ADAMTS-4) is a member of the a disintegrin and metalloprotease with thrombospondin motifs (ADAMTS) protein family that was recently identified. Aggrecanase-1 is one of two ADAMTS cartilage-degrading enzymes purified from interleukin-1-stimulated bovine nasal cartilage (Tortorella, M. D., Burn, T. C., Pratta, M. A. , Abbaszade, I., Hollis, J. M., Liu, R., Rosenfeld, S. A., Copeland, R. A., Decicco, C. P., Wynn, R., Rockwell, A., Yang, F., Duke, J. L., Solomon, K., George, H., Bruckner, R., Nagase, H., Itoh, Y., Ellis, D. M., Ross, H., Wiswall, B. H., Murphy, K., Hillman, M. C., Jr., Hollis, G. F., and Arner, E.C. (1999) Science 284, 1664-1666; 2 Abbaszade, I., Liu, R. Q., Yang, F., Rosenfeld, S. A., Ross, O. H., Link, J. R., Ellis, D. M., Tortorella, M. D., Pratta, M. A., Hollis, J. M., Wynn, R., Duke, J. L., George, H. J., Hillman, M. C., Jr., Murphy, K., Wiswall, B. H., Copeland, R. A., Decicco, C. P., Bruckner, R., Nagase, H., Itoh, Y., Newton, R. C., Magolda, R. L., Trzaskos, J. M., and Burn, T. C. (1999) J. Biol. Chem. 274, 23443-23450). The aggrecan products generated by this enzyme are found in cartilage cultures stimulated with cytokines and in synovial fluid from patients with arthritis, suggesting that aggrecanase-1 may be important in diseases involving cartilage destruction. Here we demonstrate that the thrombospondin type-1 (TSP-1) motif located within the C terminus of aggrecanase-1 binds to the glycosaminoglycans of aggrecan. Data from several studies indicate that this binding of aggrecanase-1 to aggrecan through the TSP-1 motif is necessary for enzymatic cleavage of aggrecan. 1) A truncated form of aggrecanase-1 lacking the TSP-1 motif was not effective in cleaving aggrecan. 2) Several peptides representing different regions of the TSP-1 motif effectively blocked aggrecanase-1 cleavage of aggrecan by preventing the enzyme from binding to the substrate. 3) Aggrecanase-1 was not effective in cleaving glycosaminoglycan-free aggrecan. Taken together, these data suggest that the TSP 1 motif of aggrecanase-1 is critical for substrate recognition and cleavage. PMID- 10827175 TI - TIMP-2 is required for efficient activation of proMMP-2 in vivo. AB - Matrix metalloproteinases (MMPs) are synthesized as latent proenzymes. A proteolytic cleavage event involving processing of the cysteine-rich N-terminal propeptide is required for their full activation. Previous in vitro studies indicated that activation of proMMP-2 can occur through formation of a trimolecular complex between MMP-14, TIMP-2, and proMMP-2 at the cell surface. Using TIMP-2-deficient mice and cells derived from them, TIMP-2 was shown to be required for efficient proMMP-2 activation both in vivo and in vitro. The requirement for TIMP-2 was not cell-autonomous as exogenously added TIMP-2 could restore activation of proMMP-2 to TIMP-2-deficient cells. Mutant mice were overtly normal, viable, and fertile on the C57BL/6 background, indicating that both TIMP-2 and activated proMMP-2 are dispensable for normal development. PMID- 10827176 TI - Inactivating mutation of the mouse tissue inhibitor of metalloproteinases-2(Timp 2) gene alters proMMP-2 activation. AB - To understand the biologic function of TIMP-2, a member of the tissue inhibitors of metalloproteinases family, an inactivating mutation was introduced in the mouse Timp-2 gene by homologous recombination. Outbred homozygous mutants developed and procreated indistinguishably from wild type littermates, suggesting that fertility, development, and growth are not critically dependent on TIMP-2. Lack of functional TIMP-2, however, dramatically altered the activation of proMMP 2 both in vivo and in vitro. Fully functional TIMP-2 is essential for efficient activation of proMMP-2 in vivo. No evidence of successful functional compensation was observed. The results illustrate the duality of TIMP-2 function, i.e. at low concentrations, TIMP-2 exerts a "catalytic" or enhancing effect on cell-mediated proMMP-2 activation, whereas at higher concentrations, TIMP-2 inhibits the activation and/or activity of MMP-2. PMID- 10827177 TI - X-ray crystallographic study of cyanide binding provides insights into the structure-function relationship for cytochrome cd1 nitrite reductase from Paracoccus pantotrophus. AB - We present a 1.59-A resolution crystal structure of reduced Paracoccus pantotrophus cytochrome cd(1) with cyanide bound to the d(1) heme and His/Met coordination of the c heme. Fe-C-N bond angles are 146 degrees for the A subunit and 164 degrees for the B subunit of the dimer. The nitrogen atom of bound cyanide is within hydrogen bonding distance of His(345) and His(388) and either a water molecule in subunit A or Tyr(25) in subunit B. The ferrous heme-cyanide complex is unusually stable (K(d) approximately 10(-6) m); we propose that this reflects both the design of the specialized d(1) heme ring and a general feature of anion reductases with active site heme. Oxidation of crystals of reduced, cyanide-bound, cytochrome cd(1) results in loss of cyanide and return to the native structure with Tyr(25) as a ligand to the d(1) heme iron and switching to His/His coordination at the c-type heme. No reason for unusually weak binding of cyanide to the ferric state can be identified; rather it is argued that the protein is designed such that a chelate-based effect drives displacement by tyrosine of cyanide or a weaker ligand, like reaction product nitric oxide, from the ferric d(1) heme. PMID- 10827178 TI - Feedback control of cyclooxygenase-2 expression through PPARgamma. AB - Cyclooxygenase-2 (COX-2), a rate-limiting enzyme for prostaglandins (PG), plays a key role in inflammation, tumorigenesis, development, and circulatory homeostasis. The PGD(2) metabolite 15-deoxy-Delta(12, 14) PGJ(2) (15d-PGJ(2)) was identified as a potent natural ligand for the peroxisome proliferator-activated receptor-gamma (PPARgamma). PPARgamma expressed in macrophages has been postulated as a negative regulator of inflammation and a positive regulator of differentiation into foam cell associated with atherogenesis. Here, we show that 15d-PGJ(2) suppresses the lipopolysaccharide (LPS)-induced expression of COX-2 in the macrophage-like differentiated U937 cells but not in vascular endothelial cells. PPARgamma mRNA abundantly expressed in the U937 cells, not in the endothelial cells, is down-regulated by LPS. In contrast, LPS up-regulates mRNA for the glucocorticoid receptor which ligand anti-inflammatory steroid dexamethasone (DEX) strongly suppresses the LPS-induced expression of COX-2, although both 15d-PGJ(2) and DEX suppressed COX-2 promoter activity by interfering with the NF-kappaB signaling pathway. Transfection of a PPARgamma expression vector into the endothelial cells acquires this suppressive regulation of COX-2 gene by 15d-PGJ(2) but not by DEX. A selective COX-2 inhibitor, NS-398, inhibits production of PGD(2) in the U937 cells. Taking these findings together, we propose that expression of COX-2 is regulated by a negative feedback loop mediated through PPARgamma, which makes possible a dynamic production of PG, especially in macrophages, and may be attributed to various expression patterns and physiological functions of COX-2. PMID- 10827179 TI - Distribution and repair of bipyrimidine photoproducts in solar UV-irradiated mammalian cells. Possible role of Dewar photoproducts in solar mutagenesis. AB - In order to better understand the relative contribution of the different UV components of sunlight to solar mutagenesis, the distribution of the bipyrimidine photolesions, cyclobutane pyrimidine dimers (CPD), (6-4) photoproducts ((6-4)PP), and their Dewar valence photoisomers (DewarPP) was examined in Chinese hamster ovary cells irradiated with UVC, UVB, or UVA radiation or simulated sunlight. The absolute amount of each type of photoproduct was measured by using a calibrated and sensitive immuno-dot-blot assay. As already established for UVC and UVB, we report the production of CPD by UVA radiation, at a yield in accordance with the DNA absorption spectrum. At biologically relevant doses, DewarPP were more efficiently produced by simulated solar light than by UVB (ratios of DewarPP to (6-4)PP of 1:3 and 1:8, respectively), but were detected neither after UVA nor after UVC radiation. The comparative rates of formation for CPD, (6-4)PP and DewarPP are 1:0.25 for UVC, 1:0. 12:0.014 for UVB, and 1:0.18:0.06 for simulated sunlight. The repair rates of these photoproducts were also studied in nucleotide excision repair-proficient cells irradiated with UVB, UVA radiation, or simulated sunlight. Interestingly, DewarPP were eliminated slowly, inefficiently, and at the same rate as CPD. In contrast, removal of (6-4)PP photoproducts was rapid and completed 24 h after exposure. Altogether, our results indicate that, in addition to CPD and (6-4)PP, DewarPP may play a role in solar cytotoxicity and mutagenesis. PMID- 10827180 TI - The splicing factor U1C represses EWS/FLI-mediated transactivation. AB - EWS is an RNA-binding protein involved in human tumor-specific chromosomal translocations. In approximately 85% of Ewing's sarcomas, such translocations give rise to the chimeric gene EWS/FLI. In the resulting fusion protein, the RNA binding domains from the C terminus of EWS are replaced by the DNA-binding domain of the ETS protein FLI-1. EWS/FLI can function as a transcription factor with the same DNA binding specificity as FLI-1. EWS and EWS/FLI can associate with the RNA polymerase II holoenzyme as well as with SF1, an essential splicing factor. Here we report that U1C, one of three human U1 small nuclear ribonucleoprotein specific proteins, interacts in vitro and in vivo with both EWS and EWS/FLI. U1C interacts with other splicing factors and is important in the early stages of spliceosome formation. Importantly, co-expression of U1C represses EWS/FLI mediated transactivation, demonstrating that this interaction can have functional ramifications. Our findings demonstrate that U1C, a well characterized splicing protein, can also function in transcriptional regulation. Furthermore, they suggest that EWS and EWS/FLI may function both in transcriptional and post transcriptional processes. PMID- 10827181 TI - Non-erythroid genes inserted on either side of human HS-40 impair the activation of its natural alpha -globin gene targets without being themselves preferentially activated. AB - The human alpha-globin gene complex includes three functional globin genes (5' zeta2-alpha2-alpha1-3') regulated by a common positive regulatory element named HS-40 displaying strong erythroid-specific enhancer activity. How this enhancer activity can be shared between different promoters present at different positions in the same complex is poorly understood. To address this question, we used homologous recombination to target the insertion of marker genes driven by cytomegalovirus or long terminal repeat promoters in both possible orientations either upstream or downstream from the HS-40 region into the single human alpha globin gene locus present in hybrid mouse erythroleukemia cells. We also used CRE recombinase-mediated cassette exchange to target the insertion of a tagged alpha globin gene at the same position downstream from HS-40. All these insertions led to a similar decrease in the HS-40-dependent transcription of downstream human alpha-globin genes in differentiated cells. Interestingly, this decrease is associated with the strong activation of the proximal newly inserted alpha-globin gene, whereas in marked contrast, the transcription of the non-erythroid marker genes remains insensitive to HS-40. Taken together, these results indicate that the enhancer activity of HS-40 can be trapped by non-erythroid promoters in both upstream and downstream directions without necessarily leading to their own activation. PMID- 10827182 TI - Protein kinase C-regulated dynamitin-macrophage-enriched myristoylated alanine rice C kinase substrate interaction is involved in macrophage cell spreading. AB - Macrophage spreading requires the microtubule cytoskeleton and protein kinase C (PKC). The mechanism of involvement of the microtubules and PKC in this event is not fully understood. Dynamitin is a subunit of dynactin, which is important for linking the microtubule-dependent motor protein dynein to vesicle membranes. We report that dynamitin is a Ca(2+)/calmodulin-binding protein and that dynamitin binds directly to macrophage-enriched myristoylated alanine-rice C kinase substrate (MacMARCKS), a membrane-associated PKC substrate involved in macrophage spreading and integrin activation. Dynamitin was found to copurify with MacMARCKS both during MacMARCKS purification with conventional chromatography and during the immunoabsorption of MacMARCKS using anti-MacMARCKS antibody. Vice versa, MacMARCKS was also found to cosediment with the 20 S dynactin complex. We determined that the effector domain of MacMARCKS is required to interact with the N-terminal domain of dynamitin. MacMARCKS and dynamitin also partially colocalized at peripheral regions of macrophages and in the cell-cell border of 293 epithelial cells. Treatment with phorbol esters abolished this colocalization. Disrupting the interaction with a short peptide derived from the MacMARCKS-binding domain of dynamitin caused macrophages to spread and flatten. These data suggest that the dynamitin-MacMARCKS interaction is involved in cell spreading. Furthermore, the regulation of this interaction by PKC and Ca(2+)/calmodulin provides a possible regulatory mechanism for cell adhesion and spreading. PMID- 10827183 TI - Expression of human topoisomerase I with a partial deletion of the linker region yields monomeric and dimeric enzymes that respond differently to camptothecin. AB - Human topoisomerase I is a 765-residue protein composed of four major domains as follows: the unconserved and highly charged NH(2)-terminal domain, a conserved core domain, the positively charged linker region, and the highly conserved COOH terminal domain containing the active site tyrosine. Previous studies of the domain structure revealed that near full topoisomerase I activity can be reconstituted in vitro by fragment complementation between recombinant polypeptides approximating the core and COOH-terminal domains. Here we demonstrate that deletion of linker residues Asp(660) to Lys(688) yields an active enzyme (topo70DeltaL) that purifies as both a monomer and a dimer. The dimer is shown to result from domain swapping involving the COOH-terminal and core domains of the two subunits. The monomeric form is insensitive to the anti tumor agent camptothecin and distributive during in vitro plasmid relaxation assays, whereas the dimeric form is camptothecin-sensitive and processive. However, the addition of camptothecin to enzyme/DNA mixtures causes enhancement of SDS-induced breakage by both monomeric and dimeric forms of the mutant enzyme. The similarity of the dimeric form to the wild type enzyme suggests that some structural feature of the dimer is providing a surrogate linker. Yeast cells expressing topo70DeltaL were found to be insensitive to camptothecin. PMID- 10827184 TI - RNA template-dependent 5' nuclease activity of Thermus aquaticus and Thermus thermophilus DNA polymerases. AB - DNA replication and repair require a specific mechanism to join the 3'- and 5' ends of two strands to maintain DNA continuity. In order to understand the details of this process, we studied the activity of the 5' nucleases with substrates containing an RNA template strand. By comparing the eubacterial and archaeal 5' nucleases, we show that the polymerase domain of the eubacterial enzymes is critical for the activity of the 5' nuclease domain on RNA containing substrates. Analysis of the activity of chimeric enzymes between the DNA polymerases from Thermus aquaticus (TaqPol) and Thermus thermophilus (TthPol) reveals two regions, in the "thumb" and in the "palm" subdomains, critical for RNA-dependent 5' nuclease activity. There are two critical amino acids in those regions that are responsible for the high activity of TthPol on RNA containing substrates. Mutating glycine 418 and glutamic acid 507 of TaqPol to lysine and glutamine, respectively, increases its RNA-dependent 5' nuclease activity 4-10 fold. Furthermore, the RNA-dependent DNA polymerase activity is controlled by a completely different region of TaqPol and TthPol, and mutations in this region do not affect the 5' nuclease activity. The results presented here suggest a novel substrate binding mode of the eubacterial DNA polymerase enzymes, called a 5' nuclease mode, that is distinct from the polymerizing and editing modes described previously. The application of the enzymes with improved RNA-dependent 5' nuclease activity for RNA detection using the invasive signal amplification assay is discussed. PMID- 10827185 TI - Inactivation of rabbit muscle creatine kinase by reversible formation of an internal disulfide bond induced by the fungal toxin gliotoxin. AB - The biological activity of gliotoxin is dependent on the presence of a strained disulfide bond that can react with accessible cysteine residues on proteins. Rabbit muscle creatine kinase contains 4 cysteines per 42-kDa subunit and is active in solution as a dimer. Only Cys-282 has been identified as essential for activity. Modification of this residue results in loss of activity of the enzyme. Treatment of creatine kinase with gliotoxin resulted in a time-dependent loss of activity abrogated in the presence of reducing agents. Activity was restored when the inactivated enzyme was treated with reducing agents. Inactivation of creatine kinase by gliotoxin was accompanied by the formation of a 37-kDa form of the enzyme. This oxidized form of creatine kinase was rapidly reconverted to the 42 kDa species by the addition of reducing agents concomitant with restoration of activity. A 1:1 mixture of the oxidized and reduced monomer forms of creatine kinase as shown on polyacrylamide gel electrophoresis was equivalent to the activity of the fully reduced form of the enzyme consistent with only one reduced monomer of the dimer necessary for complete activity. Conversion of the second monomeric species of the dimer to the oxidized form by gliotoxin correlated with loss of activity. Our data are consistent with gliotoxin inducing the formation of an internal disulfide bond in creatine kinase by initially binding and possibly activating a cysteine residue on the protein, followed by reaction with a second neighboring thiol. The recently published crystal structure of creatine kinase suggests the disulfide is formed between Cys-282 and Cys-73. PMID- 10827186 TI - Incorporation of nucleoside analogs into nuclear or mitochondrial DNA is determined by the intracellular phosphorylation site. AB - Nucleoside analogs used in cancer chemotherapy and in treatment of virus infections are phosphorylated in cells by nucleoside and nucleotide kinases to their pharmacologically active form. The phosphorylated nucleoside analogs are incorporated into DNA and cause cell death or inhibit viral replication. Cellular DNA is replicated both in the nucleus and in the mitochondria, and nucleoside analogs may interfere with DNA replication in both these subcellular locations. In the present study we created a cell model system where nucleoside analogs were phosphorylated, and thereby pharmacologically activated, in either the nucleus, cytosol, or mitochondria of cancer cells. The system was based on the reconstitution of deoxycytidine kinase (dCK)-deficient Chinese hamster ovary cells with genetically engineered dCK targeted to the different subcellular compartments. The nucleoside analogs phosphorylated by dCK in the mitochondria were predominantly incorporated into mitochondrial DNA, whereas the nucleoside analogs phosphorylated in the nucleus or cytosol were incorporated into nuclear DNA. We further show that the nucleoside analogs phosphorylated in the mitochondria induced cell death by an apoptotic program. These data showed that the subcellular site of nucleoside analog phosphorylation is an important determinant for incorporation of nucleoside analogs into nuclear or mitochondrial DNA. PMID- 10827187 TI - Poliovirus RNA-dependent RNA polymerase (3Dpol): structural, biochemical, and biological analysis of conserved structural motifs A and B. AB - We have constructed a structural model for poliovirus RNA-dependent RNA polymerase (3D(pol)) in complex with a primer-template (sym/sub) and ATP. Residues found in conserved structural motifs A (Asp-238) and B (Asn-297) are involved in nucleotide selection. Asp-238 appears to couple binding of nucleotides with the correct sugar configuration to catalytic efficiency at the active site of the enzyme. Asn-297 is involved in selection of ribonucleoside triphosphates over 2'-dNTPs, a role mediated most likely via a hydrogen bond between the side chain of this residue and the 2'-OH of the ribonucleoside triphosphate. Substitutions at position 238 or 297 of 3D(pol) produced derivatives exhibiting a range of catalytic efficiencies when assayed in vitro for poly(rU) polymerase activity or sym/sub elongation activity. A direct correlation existed between activity on sym/sub and biological phenotypes; a 2.5 fold reduction in polymerase elongation rate produced virus with a temperature sensitive growth phenotype. These data permit us to propose a detailed, structural model for nucleotide selection by 3D(pol), confirm the biological relevance of the sym/sub system, and provide additional evidence for kinetic coupling between RNA synthesis and subsequent steps in the virus life cycle. PMID- 10827188 TI - The inositol hexakisphosphate kinase family. Catalytic flexibility and function in yeast vacuole biogenesis. AB - Saiardi et al. (Saiardi, A., Erdjument-Bromage, H., Snowman, A., Tempst, P., and Snyder, S. H. (1999) Curr. Biol. 9, 1323-1326) previously described the cloning of a kinase from yeast and two kinases from mammals (types 1 and 2), which phosphorylate inositol hexakisphosphate (InsP(6)) to diphosphoinositol pentakisphosphate, a "high energy" candidate regulator of cellular trafficking. We have now studied the significance of InsP(6) kinase activity in Saccharomyces cerevisiae by disrupting the kinase gene. These ip6kDelta cells grew more slowly, their levels of diphosphoinositol polyphosphates were 60-80% lower than wild-type cells, and the cells contained abnormally small and fragmented vacuoles. Novel activities of the mammalian and yeast InsP(6) kinases were identified; inositol pentakisphosphate (InsP(5)) was phosphorylated to diphosphoinositol tetrakisphosphate (PP-InsP(4)), which was further metabolized to a novel compound, tentatively identified as bis-diphosphoinositol trisphosphate. The latter is a new substrate for human diphosphoinositol polyphosphate phosphohydrolase. Kinetic parameters for the mammalian type 1 kinase indicate that InsP(5) (K(m) = 1.2 micrometer) and InsP(6) (K(m) = 6.7 micrometer) compete for phosphorylation in vivo. This is the first time a PP-InsP(4) synthase has been identified. The mammalian type 2 kinase and the yeast kinase are more specialized for the phosphorylation of InsP(6). Synthesis of the diphosphorylated inositol phosphates is thus revealed to be more complex and interdependent than previously envisaged. PMID- 10827189 TI - Tryptophan 512 is sensitive to conformational changes in the rigid relay loop of smooth muscle myosin during the MgATPase cycle. AB - To examine the structural basis of the intrinsic fluorescence changes that occur during the MgATPase cycle of myosin, we generated three mutants of smooth muscle myosin motor domain essential light chain (MDE) containing a single conserved tryptophan residue located at Trp-441 (W441-MDE), Trp-512 (W512-MDE), or Trp-597 (W597-MDE). Although W441- and W597-MDE were insensitive to nucleotide binding, the fluorescence intensity of W512-MDE increased in the presence of MgADP berellium fluoride (BeF(X)) (31%), MgADP-AlF(4)(-) (31%), MgATP (36%), and MgADP (30%) compared with the nucleotide-free environment (rigor), which was similar to the results of wild type-MDE. Thus, Trp-512 may be the sole ATP-sensitive tryptophan residue in myosin. In addition, acrylamide quenching indicated that Trp-512 was more protected from solvent in the presence of MgATP or MgADP-AlF(4)( ) than in the presence of MgADP-BeF(X), MgADP, or in rigor. Furthermore, the degree of energy transfer from Trp-512 to 2'(3')-O-(N-methylanthraniloyl)-labeled nucleotides was greater in the presence of MgADP-BeF(X), MgATP, or MgADP-AlF(4)( ) than MgADP. We conclude that the conformation of the rigid relay loop containing Trp-512 is altered upon MgATP hydrolysis and during the transition from weak to strong actin binding, establishing a communication pathway from the active site to the actin-binding and converter/lever arm regions of myosin during muscle contraction. PMID- 10827190 TI - DNA repair patch-mediated double strand DNA break formation in human cells. AB - To investigate the mechanism of double strand DNA break formation in mammalian cells, an in vitro assay was established using closed circular DNA containing two uracils on opposite DNA strands (18 and 30 base pairs apart) and extracts prepared from human cells. In this assay, formation of double strand breaks was detected by the conversion of circular DNA to linear DNA. Approximately 4-fold more double strand DNA breaks were produced by extracts from cells deficient in DNA ligase I (46BR) relative to those produced by extracts from control cells (MRC5, derived from a clinically normal individual). In parallel with the amount of double strand DNA breaks, extracts from 46BR cells produced longer repair patches (up to 24 bases in length) than those from MRC5 cells (typically <5 bases long). When purified DNA ligase I was added to 46BR extracts to complement the DNA ligase deficiency, only a negligible difference was found between the amount of doublestrand DNA breaks or the repair patch size generated in the assay relative to MRC5 extracts. Together, our data demonstrate that double strand DNA breaks are produced through formation of DNA repair patches. We refer to this process of double strand break formation as the "DNA repair patch-mediated pathway." PMID- 10827191 TI - Increased AKT activity contributes to prostate cancer progression by dramatically accelerating prostate tumor growth and diminishing p27Kip1 expression. AB - The PTEN tumor suppressor gene is frequently inactivated in human prostate cancers, particularly in more advanced cancers, suggesting that the AKT/protein kinase B (PKB) kinase, which is negatively regulated by PTEN, may be involved in human prostate cancer progression. We now show that AKT activation and activity are markedly increased in androgen-independent, prostate-specific antigen positive prostate cancer cells (LNAI cells) established from xenograft tumors of the androgen-dependent LNCaP cell line. These LNAI cells show increased expression of integrin-linked kinase, which is putatively responsible for AKT activation/Ser-473 phosphorylation, as well as for increased phosphorylation of the AKT target protein, BAD. Furthermore, expression of the p27(Kip1) cell cycle regulator was diminished in LNAI cells, consistent with the notion that AKT directly inhibits AFX/Forkhead-mediated transcription of p27(Kip1). To assess directly the impact of increased AKT activity on prostate cancer progression, an activated hAKT1 mutant was overexpressed in LNCaP cells, resulting in a 6-fold increase in xenograft tumor growth. Like LNAI cells, these transfectants showed dramatically reduced p27(Kip1) expression. Together, these data implicate increased AKT activity in prostate tumor progression and androgen independence and suggest that diminished p27(Kip1) expression, which has been repeatedly associated with prostate cancer progression, may be a consequence of increased AKT activity. PMID- 10827192 TI - Assembly of U7 small nuclear ribonucleoprotein particle and histone RNA 3' processing in Xenopus egg extracts. AB - In animals, replication-dependent histone genes are expressed in dividing somatic cells during S phase to maintain chromatin condensation. Histone mRNA 3'-end formation is an essential regulatory step producing an mRNA with a hairpin structure at the 3'-end. This requires the interaction of the U7 small nuclear ribonucleoprotein particle (snRNP) with a purine-rich spacer element and of the hairpin-binding protein with the hairpin element, respectively, in the 3' untranslated region of histone RNA. Here, we demonstrate that bona fide histone RNA 3' processing takes place in Xenopus egg extracts in a reaction dependent on the addition of synthetic U7 RNA that is assembled into a ribonucleoprotein particle by protein components available in the extract. In addition to reconstituted U7 snRNP, Xenopus hairpin-binding protein SLBP1 is necessary for efficient processing. Histone RNA 3' processing is not affected by addition of non-destructible cyclin B, which drives the egg extract into M phase, but SLBP1 is phosphorylated in this extract. SPH-1, the Xenopus homologue of human p80 coilin found in coiled bodies, is associated with U7 snRNPs. However, this does not depend on the U7 RNA being able to process histone RNA and also occurs with U1 snRNPs; therefore, association of SPH1 cannot be considered as a hallmark of a functional U7 snRNP. PMID- 10827193 TI - Glycosylation/Hydroxylation-induced stabilization of the collagen triple helix. 4 trans-hydroxyproline in the Xaa position can stabilize the triple helix. AB - We have shown recently that glycosylation of threonine in the peptide Ac-(Gly-Pro Thr)(10)-NH(2) with beta-d-galactose induces the formation of a collagen triple helix, whereas the nonglycosylated peptide does not. In this report, we present evidence that a collagen triple helix can also be formed in the Ac-(Gly-Pro Thr)(10)-NH(2) peptide, if the proline (Pro) in the Xaa position is replaced with 4-trans-hydroxyproline (Hyp). Furthermore, replacement of Pro with Hyp in the sequence Ac-(Gly-Pro-Thr(beta-d-Gal))(10)-NH(2) increases the T(m) of the triple helix by 15.7 degrees C. It is generally believed that Hyp in the Xaa position destabilizes the triple helix because (Pro-Pro-Gly)(10) and (Pro-Hyp-Gly)(10) form stable triple helices but the peptide (Hyp-Pro-Gly)(10) does not. Our data suggest that the destabilizing effect of Hyp relative to Pro in the Xaa position is only true in the case of (Hyp-Pro-Gly)(10). Increasing concentrations of galactose in the solvent stabilize the triple helix of Ac-(Gly-Hyp-Thr)(10)-NH(2) but to a much lesser extent than that achieved by covalently linked galactose. The data explain some of the forces governing the stability of the annelid/vestimentiferan cuticle collagens. PMID- 10827194 TI - Elevated glucocorticoid receptor transactivation and down-regulation of alpha 1 integrin are associated with loss of plasma membrane Ca2+-ATPase isoform 1. AB - We have previously shown that inhibition of expression of the plasma membrane Ca(2+)-ATPase isoform 1 in PC6 cells leads to loss of nerve growth factor mediated neurite extension (Brandt, P.C., Sisken, J.E., Neve, R.L., and Vanaman, T.C. (1996) Proc. Natl. Acad. Sci. U.S.A. 93, 13843-13848). Cells lacking plasma membrane Ca(2+)-ATPase 1 did not attach to collagen-coated plates as tightly as controls, suggesting that a defect in adhesion might be underlying the inability to extend neurites. We report here that cell lines lacking plasma membrane Ca(2+) ATPase 1 do not produce alpha(1) integrin, which is required for both collagen adherence and neurite extension. Because alpha(1) integrin gene transcription can be down-regulated by glucocorticoids, the response of cells to glucocorticoids was investigated. Cortisol-dependent transactivation from the mouse mammary tumor virus promoter in cells lacking plasma membrane Ca(2+)-ATPase 1 was stimulated 145-216-fold over untreated cells compared with 15-26-fold for controls. This increase was not due to increased binding affinity of the receptor for cortisol, an increased number of cortisol-binding sites, or increased translocation of the receptor to the nucleus. Expression of additional glucocorticoid receptor dependent genes required for neurite extension must also be altered in cells missing the plasma membrane Ca(2+)-ATPase 1 because constitutive expression of alpha(1) integrin did not restore their nerve growth factor-mediated neurite extension capability. The impact of plasma membrane Ca(2+)-ATPase isoform 1 on other signaling systems and the resultant profound yet subtle effects on PC6 cells strongly suggests that it plays an important role in modulating signal transduction pathways downstream of Ca(2+)-mediated signals. PMID- 10827196 TI - Hdmx stabilizes Mdm2 and p53. AB - The Mdm2 protein is a key regulator of p53 activity and stability. Upon binding, Mdm2 inhibits the transcription regulatory activity of p53 and promotes its rapid degradation. In this study we investigated the effect of the human Mdm2 homologue Hdmx on p53 stability. We found that Hdmx does not target p53 for degradation, although, like Mdm2, it inhibits p53-mediated transcription activation. On the contrary, Hdmx was found to counteract the degradation of p53 by Mdm2, and to stabilize both p53 and Mdm2. The RING finger of Hdmx was found to be necessary and sufficient for this stabilization, and it probably involves hetero oligomerization with the RING finger of Mdm2, which may lead to inhibition of Mdm2's ubiquitin ligase activity. However, Hdmx does not relieve the inhibition by Mdm2 of transcription activation by p53, probably due to the formation of a trimeric complex consisting of Hdmx, Mdm2, and p53. We propose a model in which Hdmx secures a pool of largely inactive p53, which, upon the induction of stress, can be quickly activated. PMID- 10827195 TI - Cloning and characterization of a Na+-driven anion exchanger (NDAE1). A new bicarbonate transporter. AB - Regulation of intra- and extracellular ion activities (e.g. H(+), Cl(-), Na(+)) is key to normal function of the central nervous system, digestive tract, respiratory tract, and urinary system. With our cloning of an electrogenic Na(+)/HCO(3)(-) cotransporter (NBC), we found that NBC and the anion exchangers form a bicarbonate transporter superfamily. Functionally three other HCO(3)(-) transporters are known: a neutral Na(+)/ HCO(3)(-) cotransporter, a K(+)/ HCO(3)( ) cotransporter, and a Na(+)-dependent Cl(-)-HCO(3)(-) exchanger. We report the cloning and characterization of a Na(+)-coupled Cl(-)-HCO(3)(-) exchanger and a physiologically unique bicarbonate transporter superfamily member. This Drosophila cDNA encodes a 1030-amino acid membrane protein with both sequence homology and predicted topology similar to the anion exchangers and NBCs. The mRNA is expressed throughout Drosophila development and is prominent in the central nervous system. When expressed in Xenopus oocytes, this membrane protein mediates the transport of Cl(-), Na(+), H(+), and HCO(3)(-) but does not require HCO(3)(-). Transport is blocked by the stilbene 4,4' diisothiocyanodihydrostilbene- 2, 2'-disulfonates and may not be strictly electroneutral. Our functional data suggest this Na(+) driven anion exchanger (NDAE1) is responsible for the Na(+)-dependent Cl(-)-HCO(3)(-) exchange activity characterized in neurons, kidney, and fibroblasts. NDAE1 may be generally important for fly development, because disruption of this gene is apparently lethal to the Drosophila larva. PMID- 10827197 TI - The role of positively charged amino acids in ATP recognition by human P2X(1) receptors. AB - P2X receptors for ATP are a family of ligand-gated cation channels. There are 11 conserved positive charges in the extracellular loop of P2X receptors. We have generated point mutants of these conserved residues (either Lys --> Arg, Lys --> Ala, Arg --> Lys, or Arg --> Ala) in the human P2X(1) receptor to determine their contribution to the binding of negatively charged ATP. ATP evoked concentration dependent (EC(50) approximately 0.8 microm) desensitizing responses at wild-type (WT) P2X(1) receptors expressed in Xenopus oocytes. Suramin produced a parallel rightward shift in the concentration response curve with an estimated pK(B) of 6.7. Substitution of amino acids at positions Lys-53, Lys-190, Lys-215, Lys-325, Arg-202, Arg-305, and Arg-314 either had no effect or only a small change in ATP potency, time course, and/or suramin sensitivity. Modest changes in ATP potency were observed for mutants at K70R and R292K/A (20- and 100-fold decrease, respectively). Mutations at residues K68A and K309A reduced the potency of ATP by >1400-fold and prolonged the time course of the P2X(1) receptor current but had no effect on suramin antagonism. Lys-68, Lys-70, Arg-292, and Lys-309 are close to the predicted transmembrane domains of the receptor and suggest that the ATP binding pocket may form close to the channel vestibule. PMID- 10827198 TI - Micromolar Ca2+ concentrations are essential for Mg2+-dependent binding of collagen by the integrin alpha 2beta 1 in human platelets. AB - Integrin receptor alpha(2)beta(1) requires micromolar Ca(2+) to bind to collagen and to the peptide GPC(GPP)(5)GFOGER(GPP)(5)GPC (denoted GFOGER-GPP, where O represents hydroxyproline), which contains the minimum recognition sequence for the collagen-binding alpha(2) I-domain (Knight, C. G., Morton, L. F., Peachey, A. R., Tuckwell, D. S., Farndale, R. W., and Barnes, M. J. (2000) J. Biol. Chem. 275, 35-40). Platelet adhesion to these ligands is completely dependent on alpha(2)beta(1) in the presence of 2 mm Mg(2+). However, we show here that this interaction was abolished in the presence of 25 microm EGTA. Adhesion of Glanzmann's thrombasthenic platelets, which lack the fibrinogen receptor alpha(IIb)beta(3), was also inhibited by micromolar EGTA. Mg(2+)-dependent adhesion of platelets was restored by the addition of 10 microm Ca(2+), but millimolar Ca(2+) was inhibitory. Binding of isolated alpha(2)beta(1) to GFOGER GPP was 70% inhibited by 50 microm EGTA but, as with intact platelets, was fully restored by the addition of micromolar Ca(2+). 2 mm Ca(2+) did not inhibit binding of isolated alpha(2)beta(1) to collagen or to GFOGER-GPP. Binding of recombinant alpha(2) I-domain was not inhibited by EGTA, nor did millimolar Ca(2+) inhibit binding. Our data suggest that high affinity Ca(2+) binding to alpha(2)beta(1), outside the I-domain, is essential for adhesion to collagen. This is the first demonstration of a Ca(2+) requirement in alpha(2)beta(1) function. PMID- 10827199 TI - The reelin receptor ApoER2 recruits JNK-interacting proteins-1 and -2. AB - Correct positioning of neurons during embryonic development of the brain depends, among other processes, on the proper transmission of the reelin signal into the migrating cells via the interplay of its receptors with cytoplasmic signal transducers. Cellular components of this signaling pathway characterized to date are cell surface receptors for reelin like apolipoprotein E receptor 2 (ApoER2), very low density lipoprotein receptor (VLDLR), and cadherin-related neuronal receptors, and intracellular components like Disabled-1 and the nonreceptor tyrosine kinase Fyn, which bind to the intracellular domains of the ApoER2 and VLDL receptor or of cadherin-related neuronal receptors, respectively. Here we show that ApoER2, but not VLDLR, also binds the family of JNK-interacting proteins (JIPs), which act as molecular scaffolds for the JNK-signaling pathway. The ApoER2 binding domain on JIP-2 does not overlap with the binding sites for MLK3, MKK7, and JNK. These results suggest that ApoER2 is able to assemble a multiprotein complex containing Disabled-1 and JIPs, together with their binding partners, to the cell surface of neurons. This complex might participate in ApoER2-specific reelin signaling and thus would explain the different phenotype of mice lacking the ApoER2 from that of VLDLR-deficient mice. PMID- 10827200 TI - The assembly of very low density lipoproteins in rat hepatoma McA-RH7777 cells is inhibited by phospholipase A2 antagonists. AB - In McA-RH7777 cells, the oleate-stimulated assembly and secretion of very low density lipoproteins (VLDL) was associated with enhanced deacylation of phospholipids, which was markedly decreased by inactivation of the cellular phospholipase A(2). Treatment of the cells with antagonists or antisense oligonucleotide of the Ca(2+)-independent phospholipase A(2) (iPLA(2)) significantly inhibited secretion of apoB100-VLDL and triglyceride. Similar inhibitory effect of the iPLA(2) antagonists was observed on apoB48-VLDL secretion, but secretion of high density lipoprotein particles (such as apoAI- and apoB48-high density lipoprotein) or proteins in general was unaffected. The iPLA(2) antagonist did not affect the synthesis of apoB100 or triglyceride, nor did it affect the activities of phospholipase D, phosphatidate phosphohydrolase, or microsomal triglyceride transfer protein. Inactivation of iPLA(2) resulted in impaired apoB100-VLDL assembly as shown by decreased apoB100-VLDL and triglyceride within the microsomal lumen, with concomitant increase in apoB100 association with the microsomal membranes. The inhibitory effect of iPLA(2) antagonists on apoB100-VLDL assembly/secretion could be abated by pretreatment of cells with oleate. Analysis of molecular species of microsomal phosphatidylcholine and phosphatidylethanolamine by electron spray tandem mass spectrometry revealed that the enrichment of oleoyl moieties was altered by the treatment of iPLA(2) antagonist. These results suggest that the oleate-induced VLDL assembly/secretion may depend upon the establishment of membrane glycerolipids enriched in oleoyl chain, a process mediated by the iPLA(2) activity. PMID- 10827201 TI - Processing by endoplasmic reticulum mannosidases partitions a secretion-impaired glycoprotein into distinct disposal pathways. AB - In the early secretory pathway, a distinct set of processing enzymes and family of lectins facilitate the folding and quality control of newly synthesized glycoproteins. In this regard, we recently identified a mechanism in which processing by endoplasmic reticulum mannosidase I, which attenuates the removal of glucose from asparagine-linked oligosaccharides, sorts terminally misfolded alpha(1)-antitrypsin for proteasome-mediated degradation in response to its abrogated physical dissociation from calnexin (Liu, Y., Choudhury, P., Cabral, C., and Sifers, R. N. (1999) J. Biol. Chem. 274, 5861-5867). In the present study, we examined the quality control of genetic variant PI Z, which undergoes inappropriate polymerization following biosynthesis. Here we show that in stably transfected hepatoma cells the additional processing of asparagine-linked oligosaccharides by endoplasmic reticulum mannosidase II partitions variant PI Z away from the conventional disposal mechanism in response to an arrested posttranslational interaction with calnexin. Intracellular disposal is accomplished by a nonproteasomal system that functions independently of cytosolic components but is sensitive to tyrosine phosphatase inhibition. The functional role of ER mannosidase II in glycoprotein quality control is discussed. PMID- 10827202 TI - Distinct NMDA receptor subpopulations contribute to long-term potentiation and long-term depression induction. AB - Long-term potentiation (LTP) and long-term depression (LTD) are persistent modifications of synaptic strength that have been implicated in learning, memory, and neuronal development. Despite their opposing effects, both forms of plasticity can be triggered by the activation of NMDA receptors. One mechanism proposed for this bidirectional response is that the specific patterns of afferent stimulation producing LTP and LTD activate to different degrees a uniform receptor population. A second possibility is that these patterns activate separate receptor subpopulations composed of different NMDA receptor (NR) subunits. To test this hypothesis we examined the inhibition of LTP and LTD by a series of competitive NMDA receptor antagonists that varied in their affinities for NR2A/B and NR2C/D subunits. The potency for the inhibition of LTP compared with inhibition of LTD varied widely among the agents. Antagonists with higher affinity for NR2A/B subunits relative to NRC/D subunits showed more potent inhibition of LTP than of LTD. D-3-(2-carboxypiperazine-4-yl)-1-propenyl-1 phosphonic acid, which binds to NR2A/B with very high affinity relative to NR2C/D, showed an approximately 1000-fold higher potency for LTP than for LTD. These results show that distinct subpopulations of NMDA receptors characterized by different NR2 subunits contribute to the induction mechanisms of potentiation and depression. PMID- 10827203 TI - Thomas Hughes Jukes (1906-1999). PMID- 10827204 TI - Cyclic fatty acid monomers from heated oil modify the activities of lipid synthesizing and oxidizing enzymes in rat liver. AB - Cyclic fatty acid monomers purified from a heated linseed oil were given for 2 wk to adult rats as triacylglycerol at two dose levels, i.e., 0.1 and 1 g/100 g diet, to determine their effect on some aspects of lipid metabolism. Indirect evidence of a peroxisome proliferator-like effect was observed, as determined by an elevation of some characteristic enzyme activities, such as peroxisomal acyl CoA oxidase, and the microsomal omega- but also (omega-1)-laurate hydroxylase (CYP4A1 and CYP2E1, respectively). The dietary cyclic fatty acids induced a coordinated regulation between the activities of the lipogenic enzymes studied (Delta9-desaturase, phosphatidate phosphohydrolase) and peroxisomal oxidation, but not with mitochondrial beta-oxidation. The dose-dependent decrease of Delta9 desaturase activity (P < 0.05) with cyclic fatty acid monomer intake was accompanied by a similar decrease of the monounsaturated fatty acid level in liver. The increase in the gamma-linolenic acid level also suggested an increase in Delta6-desaturase activity with cyclic fatty acid intake (P < 0.05). In addition, our results strongly suggested that the altered liver levels of eicosapentaenoic and arachidonic acids were due to the peroxisomal retroconversion process in rats fed cyclic acids. Finally, an effect of these cyclic compounds on the carbohydrate metabolism cannot be disregarded because they decreased liver glycogen concentration. We conclude that cyclic fatty acid monomers affect different aspects of lipid metabolism, including a phenotypic peroxisome proliferator response. This provides the ground for further studies investigating the biochemical pathways that underlie the nutritional effect of such molecules. PMID- 10827206 TI - Copper deficiency suppresses effector activities of differentiated U937 cells. AB - Dietary copper (Cu) deficiency impairs both innate and acquired branches of immunity. Specific roles of Cu in the activation and effector activities of host defense cells remain largely unknown. The effects of Cu status on effector activities of a monocytic cell line were investigated as an initial step in the elucidation of specific functions of Cu in phagocytic cells. Exposure of differentiating U937 human promonocytic cells to 5 micromol/L 2,3, 2-tetraamine (tet), a high affinity Cu chelator, for 4 d decreased cellular Cu by 62% without altering cellular Cu,Zn-superoxide dismutase (SOD) activity, Zn content, mitochondrial activity and protein synthesis. In contrast, Cu deficiency suppressed the respiratory burst activity and markedly compromised the ability of U937 cells to kill Salmonella. Similarly, treatment of RAW264.7 murine macrophages with 5 micromol/L tet decreased cell Cu by 78% and Cu,Zn-SOD activity by 15% and increased bacterial survival by 180%. The tet-induced impairment of respiratory burst and bactericidal activities was blocked in cultures supplemented with Cu, but not Zn or Fe. In addition, lipopolysaccharide (LPS) induced secretion of the inflammatory mediators, tumor necrosis factor-alpha, interleukin (IL)-1beta, IL-6 and prostaglandin E(2) (PGE(2)), was decreased by 30 60% in tet-treated U937 cells. Flow cytometric analysis of the surface antigens CD11b and CD71 showed that the suppressed activities of Cu-deficient cells were not due to an attenuation in the degree of differentiation or secondary iron deficiency. These data demonstrate that U937 cells provide a useful model for examining the biochemical roles of Cu in monocyte activity. PMID- 10827205 TI - A high fructose diet affects the early steps of insulin action in muscle and liver of rats. AB - A high fructose diet induces insulin resistance in rats, although the exact molecular mechanism involved is unknown. In this study, we used immunoprecipitation and immunoblotting to examine the levels and phosphorylation status of the insulin receptor (IR) and insulin receptor substrate-1 (IRS-1), as well as the association of the IRS-1 with phosphatidylinositol 3-kinase (PI 3 kinase), and phosphotyrosine phosphatase (SHP2) in the liver and muscle of rats fed a control or high fructose diet for 28 d. There were no differences in IR and the IRS-1 protein levels in the liver and muscle of rats fed the control and high fructose diets. However, tyrosine-phosphorylation of the insulin receptor after insulin stimulation was reduced to 71 +/- 2% (P < 0.05) of control in the liver of the fructose-fed rats. In samples previously immunoprecipitated with anti-IRS 1 antibody and blotted with antiphosphotyrosine antibody, the insulin-stimulated IRS-1 phosphorylation levels in the liver and muscle of the fructose-fed group were only 70 +/- 6% (P < 0.05) and 76 +/- 5% (P < 0.05) of those of control rats, respectively. The insulin-stimulated IRS-1 association with PI 3-kinase was reduced to 84 +/- 3% (P < 0.05) in the liver and to 84 +/- 4% (P < 0.05) in the muscle of the fructose-fed group compared with control rats. Insulin-stimulated IRS-1 association with SHP2 was reduced to 79 +/- 5% (P < 0.05) in liver of the fructose-fed rats. These data suggest that changes in the early steps of insulin signal transduction may have an important role in the insulin resistance observed in these rats. PMID- 10827207 TI - Hyperparathyroidism is augmented by ovariectomy in Nagase analbuminemic rats. AB - The role of albumin in bone metabolism was studied in Nagase analbuminemic (NA) rats. Serum calcium (Ca), inorganic phosphate (Pi) and magnesium (Mg) concentrations did not differ between female NA and control Sprague-Dawley (SD) rats at the time of ovariectomy (ovx), although serum ionized Ca was significantly lower in NA rats than in SD rats. Serum parathyroid hormone (PTH) and osteocalcin (OC) concentrations and urinary Ca excretion were significantly greater in NA rats than in SD rats, suggesting hyperparathyroidism and the resultant enhanced bone turnover in NA rats. Paradoxically, ovx increased serum PTH and OC in NA rats but not in SD rats. Ovx-induced exacerbation of hyperparathyroidism was confirmed by significantly greater conversion of 25 hydroxyvitamin D to 1, 25-dihydroxyvitamin D in ovx NA rats even after normalization to vitamin D-binding protein. Bone mineral density (BMD) in proximal tibia increased similarly in a time-dependent manner in sham-operated NA and SD rats. However, ovx ablated the time-dependent increase of BMD in SD rats and significantly decreased BMD in NA rats by 2 wk after ovx, resulting in a significantly lower BMD in ovx NA rats than in ovx SD rats. In summary, NA rats, which are analbuminemic with compensatory increases in lipid and protein synthesis, developed hyperparathyroidism, possibly due to an increase in serum Pi and a reduction of ionized Ca, and ovx induced a greater BMD reduction in NA rats than in SD rats, probably by exacerbating hyperparathyroidism. PMID- 10827208 TI - Dietary conjugated linoleic acid reduces rat adipose tissue cell size rather than cell number. AB - We investigated the basis for the reduction in fat pad size in rats fed conjugated linoleic acid (CLA). In the first study, growing female Sprague-Dawley rats (initial weight150 g) were fed diets containing 0, 0.25 and 0.5 g/100 g diet of a purified (97% CLA) and 0.5% of a feed-grade (55% CLA) source of CLA for 5 wk to determine the effects on growth performance and fat mass. There was no effect of CLA on growth rate or food intake. Dietary CLA reduced retroperitoneal fat pad weight 13, 25 and 32% in rats fed 0.25 and 0. 5% of the pure CLA and 0.5% of the feed-grade CLA, respectively (P < 0.05). Similar effects were observed in the parametrial fat pad. The reduced pad size was due to smaller adipocyte size rather than a reduced cell number. Relative to the control group, mean cell volume was 15, 28 and 29% lower in tissue from rats fed 0.25 and 0.5% of the pure CLA and 0.5% of the feed-grade CLA, respectively (P < 0.01). In the second study, rats were fed CLA (0 vs. 0.5%) for 7 or 49 d. Reductions in fat pad weight were observed within 7 d. In addition, the effects of CLA on energy metabolism were studied in the chronically fed rats. There were no significant effects of CLA on oxygen consumption, CO(2) or heat production. During wk 4 of feeding, but not at other times, there was a 5% lower respiratory quotient in CLA-fed rats (P < 0.05). There was a time-dependent accumulation of CLA in adipose tissue and a decrease in monounsaturated fatty acids. These results suggest that the reduction in fat mass in rats fed CLA can be accounted for by a reduction in cell size rather than a change in cell number. PMID- 10827209 TI - Evidence for multiple signaling pathways in the regulation of gene expression by amino acids in human cell lines. AB - In mammals, plasma concentrations of amino acids (AA) are affected by nutritional or pathologic conditions. Alterations in AA profiles have been reported as a result of a deficiency of any one of the essential AA, a dietary imbalance of AA or an insufficient intake of protein. In recent years, evidence has accumulated that AA availability regulates the expression of several genes involved in the regulation of a number of cellular functions or AA metabolism. Nevertheless, the molecular mechanisms involved in the AA regulation of mammalian gene expression are limited, particularly the signaling pathways mediating the AA response. This work provides a better understanding of the signaling pathways involved in the AA control of gene expression. We studied the expression of C/EBP homologous protein (CHOP) and asparagine synthetase (AS) in response to deprivation of a single AA and investigated the possible link between protein synthesis inhibition due to amino acid limitation and gene expression. We have shown the following: 1) several mechanisms are involved in the AA control of gene expression. When omitted from the culture medium, each AA can activate one (or several) specific signaling pathways leading to the regulation of one specific pattern of genes. 2) AA limitation by itself can induce gene expression independently of a cellular stress due to protein synthesis inhibition. Together, these results suggest that AA control of gene expression involves several specific mechanisms by which one AA (or one group of AA) can activate one signaling pathway and thus alter one specific pattern of gene expression. PMID- 10827210 TI - Regulation of hepatic delta-6 desaturase expression and its role in the polyunsaturated fatty acid inhibition of fatty acid synthase gene expression in mice. AB - Dietary polyunsaturated fatty acids (PUFA) of the (n-6) and (n-3) families uniquely suppress the expression of lipogenic genes while concomitantly inducing the expression of genes encoding proteins of fatty acid oxidation. Although considerable progress has been made toward understanding the nuclear events affected by PUFA, the intracellular mediator responsible for the regulation of hepatic lipogenic gene expression remains unclear. On the basis of earlier fatty acid composition studies, we hypothesized that the Delta-6 desaturase pathway was essential for the production of the fatty acid regulator of gene expression. To address this hypothesis, male BALB/c mice (n = 8/group) were fed for 5 d a high glucose, fat-free diet (FF) or the FF plus 50 g/kg 18:2(n-6) with and without eicosa-5, 8,11,14-tetraynoic acid (ETYA) (200 mg/kg diet), a putative inhibitor of the Delta-6 desaturase pathway. ETYA had no effect on food intake or weight gain, but it completely prevented 18:2(n-6) from suppressing the hepatic abundance of fatty acid synthase mRNA. ETYA ingestion was associated with a decrease in the hepatic content of 20:4(n-6) and an increase in the amount of 18:2(n-6). The fatty acid composition changes elicited by ETYA were accompanied by a decrease in the enzymatic activity of Delta-6 desaturase. Interestingly, the hepatic abundance of Delta-6 desaturase mRNA was actually induced by ETYA one- to twofold. When the product of Delta-6 desaturase, i.e., 18:3(n-6), was added to the ETYA plus 18:2(n-6) diet, the hepatic content of 20:4(n-6) was normalized. In addition, 18:3(n-6) consumption reduced the level of hepatic Delta-6 desaturase mRNA by 50% and completely prevented the increase in fatty acid synthase mRNA that was associated with ETYA ingestion. Apparently, Delta-6 desaturation is an essential step for the PUFA regulation of the fatty acid synthase gene transcription. Finally, the suppression of Delta-6 desaturase by PUFA and its induction by ETYA suggest that the Delta-6 desaturase gene may be regulated by two different lipid-dependent mechanisms. PMID- 10827211 TI - Glutamine appearance rate in plasma is not increased after gastrointestinal surgery in humans. AB - The metabolic response to surgical stress is characterized by muscle protein breakdown and mobilization of amino acids and has been postulated to furnish glutamine and other amino acids to the immune system, gut and liver. The present study was undertaken to investigate whether the whole body appearance rate (R(a))(3) of glutamine in plasma is increased after major elective surgery. Fourteen patients (8 males, 6 females) were measured prior to laparotomy and on the second postoperative day. Patients received a primed continuous 6-h infusion of L-[5-(15) N]glutamine and L-[1-(13)C]leucine, and arterial blood samples and muscle biopsies were taken for concentration and enrichment measurements. As expected, the metabolic response to surgery was characterized by a rise in whole body protein breakdown (n = 14, P < 0.001) and a decreased concentration of glutamine in plasma (n = 14, P < 0.001) and muscle (n = 8, P < 0.01). However, these catabolic changes were not reflected by an increase in the plasma R(a) of glutamine: 246 +/- 8 micromol. kg(-1). h(-1) before surgery vs. 241 +/- 10 micromol. kg(-1). h(-1) on the second postoperative day. We conclude that the whole body R(a) of glutamine in plasma is not increased 2 d after elective gastrointestinal surgery. Further studies are warranted to establish whether the lack of an increase in plasma glutamine R(a) provides a rationale for glutamine supplementation. PMID- 10827212 TI - Chronic consumption of short-chain fructooligosaccharides does not affect basal hepatic glucose production or insulin resistance in type 2 diabetics. AB - Short-chain fructooligosaccharides (FOS) are prebiotics, which escape digestion in the small intestine and are fermented by the colonic microflora into short chain fatty acids. Recently, we found that the daily consumption of 20 g FOS decreased basal hepatic glucose production in healthy subjects without any effect on insulin-stimulated glucose metabolism. In this study, we evaluated the effects of the chronic ingestion of FOS on plasma lipid and glucose concentrations, hepatic glucose production and insulin resistance in type 2 diabetics. Type 2 diabetic volunteers (n = 10; 6 men, 4 women) received either 20 g/d FOS or sucrose for 4 wk in a double-blind crossover design. FOS did not modify fasting plasma glucose and insulin concentrations or basal hepatic glucose production. The plasma glucose response to a fixed exogenous insulin bolus did not differ at the end of the two periods. Erythrocyte insulin binding also did not differ. Serum triacylglycerol, total and HDL cholesterol, free fatty acid, apolipoproteins A1 and B and lipoprotein (a) concentrations were not modified by the chronic ingestion of FOS. We conclude that 4 wk of 20 g/d of FOS had no effect on glucose and lipid metabolism in type 2 diabetics. PMID- 10827213 TI - Fruits and vegetables increase plasma carotenoids and vitamins and decrease homocysteine in humans. AB - Observational epidemiologic studies have shown that a high consumption of fruits and vegetables is associated with a decreased risk of chronic diseases. Little is known about the bioavailability of constituents from vegetables and fruits and the effect of these constituents on markers for disease risk. Currently, the recommendation is to increase intake of a mix of fruits and vegetables ("five a day"). We investigated the effect of this recommendation on plasma carotenoids, vitamins and homocysteine concentrations in a 4-wk dietary controlled, parallel intervention study. Male and female volunteers (n = 47) were allocated randomly to either a daily 500-g fruit and vegetable ("high") diet or a 100-g fruit and vegetable ("low") diet. Analyzed total carotenoid, vitamin C and folate concentrations of the daily high diet were 13.3 mg, 173 mg and 228.1 microg, respectively. The daily low diet contained 2.9 mg carotenoids, 65 mg vitamin C and 131.1 microg folate. Differences in final plasma levels between the high and low group were as follows: lutein, 46% [95% confidence interval (CI) 28-64]; beta cryptoxanthin, 128% (98-159); lycopene, 22% (8-37); alpha-carotene, 121% (94 149); beta-carotene, 45% (28-62); and vitamin C, 64% (51-77) (P < 0.05). The high group had an 11% (-18 to -4) lower final plasma homocysteine and a 15% (0.8-30) higher plasma folate concentration compared with the low group (P < 0.05). This is the first trial to show that a mix of fruits and vegetables, with a moderate folate content, decreases plasma homocysteine concentrations in humans. PMID- 10827214 TI - Folate status of elderly women following moderate folate depletion responds only to a higher folate intake. AB - Dietary Reference Intakes (DRI) for folate for elderly women have been based primarily on data extrapolated from studies in younger women. This study was conducted to provide the first age-specific data in elderly women (60-85 y) from a controlled metabolic study on which to base folate intake recommendations. Subjects (n = 33) consumed a moderately folate-deplete (118 microg/d) diet for 7 wk, followed by repletion diets providing either 200 or 415 microg folate/d as diet plus folic acid (FA) or a combination of FA and orange juice (OJ) for 7 wk (n = 30). Comparisons among and within groups were made for serum folate (SF), RBC folate and plasma total homocysteine (tHcy) concentrations. SF concentrations decreased significantly (P < 0.001) during depletion (65 +/- 15%). Postrepletion, the adjusted SF concentration for subjects consuming 415 microg folate/d was significantly greater (P = 0.003) than for subjects consuming 200 microg folate/d. RBC folate concentrations decreased (P < 0.001) during depletion (21 +/ 10%) and further (P < 0.001) during repletion (5 +/- 14%). During depletion, plasma tHcy concentrations increased significantly (P < 0.001) and an inverse relationship between SF and plasma tHcy concentrations was observed in 94% of subjects (P < 0.001). Reversal of this inverse relationship was significant only for subjects consuming 415 microg folate/d (P < 0.001). Postrepletion, subjects consuming 200 microg folate/d had a significantly higher (P = 0.009) adjusted plasma tHcy concentration than subjects consuming 415 microg folate/d. These data in elderly women indicate that 415 microg/d folate, provided as a combination of diet, FA and OJ, or diet and FA, normalizes folate status more effectively than does 200 microg/d, thus providing age-specific data for future folate intake recommendations. PMID- 10827215 TI - LDL of Taiwanese vegetarians are less oxidizable than those of omnivores. AB - The vegetarians in Taiwan consume diets high in polyunsaturated fatty acids. To investigate whether this dietary pattern results in high susceptibility of LDL to oxidation, 109 long-term (8 +/- 5 y) male and female vegans and lactovegetarians (ages 31-45 y) from Taipei and females from Hualien and matched omnivores were recruited to have 24-h-recall dietary assessments and blood lipid analysis. Body mass index and blood pressure were significantly lower in all vegetarian groups than in the matched omnivore groups (P < 0.05). Vegetarians consumed less energy except in the males and less protein, fat and cholesterol (P < 0.05). The mean polyunsaturated/saturated fatty acid (P/S) ratio of 2.4 in vegetarian diet was about two times that in omnivore diet (P < 0. 001). The concentrations of plasma total- and LDL-cholesterol (LDL-C) but not HDL-cholesterol (HDL-C) were significantly lower (P < 0.001) and resulting HDL-C/LDL-C ratio was 38, 46 and 30% higher (P < 0.01) in Taipei female, male and Hualien female vegetarians, respectively, than in the matched omnivores. Plasma triglyceride concentration was significantly lower only in the Hualien women vegetarians (31%, P < 0.001) than in the matched omnivores. The lag time of conjugated diene formation in LDL oxidized in vitro induced by copper was longer in Taipei female (62%, P < 0.001), male (29%, P < 0.05) and Hualien female (38%, P < 0.01), and the production of thiobarbituric acid reactive substances (TBARS) in LDL after 2-4 h of oxidation was 22-32% less (P < 0.005) in Taipei male and Hualien female vegetarians than the matched omnivores. Lag time of LDL oxidation was negatively related to LDL arachidonic (r = -0.55, P = 0.0003) and eicosapentaenoic (r = -0.47, P = 0.003) acid contents. LDL-TBARS production was negatively related to LDL linoleic acid content (r = -0.36, P = 0.023), but positively related to LDL arachidonic (r = 0.56, P = 0.0002) and eicosapentaenoic (r = 0.45, P = 0.004) acids. No significant differences were found in dietary vitamins C and E intakes and plasma LDL alpha-tocopherol concentrations between vegetarians and omnivores. Our results suggest that vegetarian diets decrease the susceptibility of LDL to oxidation despite their higher dietary P/S ratio. PMID- 10827216 TI - Poor maternal schooling is the main constraint to good child care practices in Accra. AB - Life in urban areas presents special challenges for maternal child care practices. Data from a representative quantitative survey of households with children < 3 y of age in Accra, Ghana were used to test a number of hypothesized constraints to child care including various maternal (anthropometry, education, employment, marital status, age and ethnic group) and household-level factors (income, availability of food, quality of housing and asset ownership, availability of services, household size and crowding). Three care indices were created as follows: 1) a child feeding index; 2) a preventive health seeking index; and 3) a hygiene index. The first two indices were based on data from maternal recall; the hygiene index was based on spot-check observations of proxies of hygiene behaviors. Multivariate analyses (ordinary least-squares regression for the child feeding index and ordered probit for the two other indices) showed that maternal schooling was the most consistent constraint to all three categories of child care practices. None of the household-level characteristics were associated with child feeding practices, but household socioeconomic factors were associated with better preventive health seeking and hygiene behaviors. Thus, poor maternal schooling was a main constraint for child feeding, health seeking and hygiene practices in Accra, but the lack of household resources was a constraint only for health seeking and hygiene. The programmatic implications of these findings for interventions in nutrition education and behaviors in Accra are discussed. PMID- 10827217 TI - The cecum and dietary short-chain fructooligosaccharides are involved in preventing postgastrectomy anemia in rats. AB - Dietary short-chain fructooligosaccharides (Sc-FOS) stimulate absorption of calcium and magnesium in the large intestine of rats. In this study, we examined whether Sc-FOS stimulate iron absorption in the large intestine by monitoring recovery from anemia in gastrectomized rats, with or without cecectomy. The rats were divided into four groups, i.e., sham-operated (Sham), gastrectomized only (GX), cecectomized only (CX) and both gastrectomized and cecectomized (GCX). Half of the rats in each group were fed a control diet (AIN-93G) and the other half were fed a Sc-FOS-containing diet (75 g/kg diet) for 28 d. Hematocrit (Ht) and hemoglobin concentration (Hb) were measured at the start, and on d 14 and 28 after the start of feeding. On the final day of the study, total blood was collected. Gastrectomy significantly decreased Ht and Hb, but cecectomy did not influence these variables. Dietary Sc-FOS prevented the decrease in Ht and Hb significantly. In the gastrectomized rats, the effectiveness of Sc-FOS in preventing postgastrectomy anemia was significantly diminished by cecectomy. These results suggest that the effect of Sc-FOS in increasing absorption of iron in gastrectomized rats takes place in part in the cecum. The cecum plays an important role in the mechanism by which Sc-FOS prevent postgastrectomy anemia. PMID- 10827218 TI - Tissue lycopene concentrations and isomer patterns are affected by androgen status and dietary lycopene concentration in male F344 rats. AB - Diets rich in lycopene from tomato products as well as greater concentrations of blood lycopene have been associated with a decreased risk for prostate cancer in epidemiologic studies. However, little is known about factors modulating lycopene absorption, metabolism and tissue distribution in humans and animal models of prostate cancer. A 2 x 4 factorial design was used to measure the effects of androgen status (castrated vs. intact), dietary lycopene concentration (0.00-5.00 g/kg lycopene) and their interaction on tissue lycopene accumulation and isomer patterns in male F344 rats. Male F344 rats ( 14 wk old; 44 castrated, 44 intact) were randomly assigned to one of four diets containing total lycopene concentrations of 0.00, 0.05, 0.50 or 5.00 g/kg as beadlets and fed for 8 wk. Tissue total lycopene and cis/trans lycopene profiles were determined by HPLC. Tissue and serum lycopene concentrations increased significantly (P < 0.01) as dietary lycopene levels increased between 0.00 and 0.50 g/kg. No further increases in serum or tissue concentrations were seen in rats fed dietary lycopene between 0.50 and 5.00 g/kg. As dietary lycopene increased, so did the percentage of cis lycopene in the liver (P < 0.05), due primarily to an increase in the 5-cis isomer. Castrated rats accumulated twice (P < 0.01) the liver lycopene as compared to intact controls, with no effect of castration on serum lycopene or adrenal, kidney, adipose, or lung tissue concentration. Livers from castrated rats had a greater proportion of cis-lycopene than those of intact rats (P < 0.05). A significant interaction between dietary lycopene concentration and androgen status was seen for liver lycopene concentration (P < 0.01). We conclude that serum and tissue lycopene reaches a plateau between 0.05 and 0.50 g/kg dietary lycopene, the tissue cis/trans lycopene ratio increases with greater dietary lycopene and androgens modulate hepatic lycopene metabolism. PMID- 10827219 TI - Lymphatic transport in rats of several dietary fats differing in fatty acid profile and triacylglycerol structure. AB - We examined in rats the intestinal absorption of nine very different dietary fats (two rapeseed oils, corn, olive, palm and menhaden oil, butter, cocoa butter and lard) to investigate the influence of fatty acid profile and triacylglycerol structure on absorption. Absorption was followed for 24 h after administration of similar amounts of fats, and the accumulated lymphatic transport and amount of triacylglycerols found in lymph in response to the administered fats were calculated, revealing major differences. The transport of olive and low alpha linolenic rapeseed oil was significantly higher than that of the other fats (P < 0.05), except corn oil. The lymphatic transport of the other fats followed a slower course, with cocoa butter and menhaden oil having the lowest amounts transported. The amount of triacylglycerols found in lymph in response to the administered fats at 8 h ranged from 27.5% of the administered dose for cocoa butter to 72.1% for olive oil. The value for cocoa butter was significantly lower than that for most other fats. At 24 h, the values ranged from 66.5% for cocoa butter to 121.2% for olive oil. The high value for olive oil suggested transport of endogenous as well as exogenous fatty acids. The low value observed after cocoa butter resulted from decreased lipolysis and possibly also low absorption of triacylglycerols with high levels of long-chain saturated fatty acids in the sn-1/3 position. Furthermore, a low value was observed after menhaden oil administration, suggesting decreased absorption of fats containing (n-3) long chain polyunsaturated fatty acids. Overall, these results demonstrate the influence of the fatty acid composition and triacylglycerol structure on the lymphatic absorption of dietary fat. PMID- 10827220 TI - Thiamin is decomposed due to Anaphe spp. entomophagy in seasonal ataxia patients in Nigeria. AB - A fairly high activity of a relatively heat-resistant thiaminase was detected and characterized from the pupae of an African silkworm Anaphe spp. which had been the putative cause of a seasonal ataxia and impaired consciousness in Nigerians. The thiaminase in the buffer extract of Anaphe pupae was type I (thiamin: base 2 methyl-4-aminopyrimidine methyl transferase EC 2.5.1.2), and the optimal temperature and pH were 70 degrees C and 8.0-8.5, respectively. Based on gel filtration chromatography, the molecules were estimated to be 200 kDa. Second substrates which could be utilized by the thiaminase were pyridoxine, amino acids, glutathione, taurine and 4-aminopyridine. Thiamin phosphate esters were inactive as substrates. This is the first report describing an insect thiaminase. Our results indicate the necessity of thorough heat treatment for the detoxification of the African silkworm, making the worm a safe source of high quality protein. PMID- 10827221 TI - Intakes of dietary docosahexaenoic acid ethyl ester and egg phosphatidylcholine improve maze-learning ability in young and old mice. AB - The effect of dietary docosahexaenoic acid (DHA) [22:6 (n-3)] ethyl ester (EE) and egg-phosphatidylcholine (PC) on maze-learning ability in young and old mice was studied. Male Crj:CD-1 mice aged either 3 wk or 14 mo were fed a diet containing 2 g DHA-EE/100 g diet plus 3 g palm oil/100 g diet (DHA-EE Group), 5 g egg-PC/100 g diet (egg-PC Group), 1 g DHA-EE/100 g diet plus 2.5 g egg-PC/100 g diet plus 1.5 g palm oil/100 g diet (DHA-EE + egg-PC Group) or 5 g palm oil/100 g diet (Control Group) for 5 mo. Maze-learning ability was assessed 4 mo after the start of the experiment. The time required to reach the maze exit and the number of times that a mouse strayed into blind alleys in the maze were measured in three trials every 4 d. In trial 2 of young mice, performed on d 4 after the first trial, the DHA-EE group required less (P < 0.05) time to reach the maze exit and DHA-EE and egg-PC groups strayed (P < 0.05) into blind alleys fewer times than the control group. In trial 2 of old mice, the DHA-EE, egg-PC and DHA EE + egg-PC groups needed less (P < 0.05) time to find the exit and spent a fewer (P < 0.05) number of times in blind alleys than did the control group. The DHA EE, DHA-EE + egg-PC and egg-PC groups strayed into blind alleys fewer times than the control group in trial 3 of old mice (P < 0.05). Our results suggest that the intake of DHA-EE and the egg-PC diet effectively enhances maze-learning ability and brain functions in old mice. PMID- 10827222 TI - Supplementation with L-histidine during dietary zinc repletion improves short term memory in zinc-restricted young adult male rats. AB - Zinc, an essential dietary element, modulates neurotransmission in brain regions associated with cognition. Cognitive dysfunction has been reported in offspring of female rats fed zinc-restricted diets during gestation and/or lactation. Studies on the cognitive effects of zinc restriction during young adulthood are limited. After a 3-wk period of dietary zinc restriction, male rats (71-75 d old) were repleted with zinc chloride alone, or zinc chloride supplemented with L histidine, and short-term memory was measured using the Morris water maze. During restriction, zinc-restricted rats demonstrated significantly longer (86.0%) retrieval latencies than nonrestricted controls, and significantly lower liver (25.5%), bone (32.5%) and hippocampal (3.2%) zinc concentrations. During subsequent repletion, rats repleted with zinc chloride supplemented with L histidine improved their retrieval latencies to the extent that they were no longer significantly different from controls by repletion d 3. This was associated with a return of hippocampal zinc concentrations to control values by repletion d 3. The mean retrieval escape latencies of the zinc chloride-repleted rats remained significantly prolonged (75.0%). Collectively, these data indicate the following: 1) feeding a zinc-restricted diet for 3 wk impairs short-term memory in young adult male rats, and 2) repletion with dietary zinc supplemented with L-histidine improves short-term memory function more efficiently than dietary zinc chloride alone. The latter point suggests that dietary zinc supplemented with L-histidine is more bioavailable to the brain than zinc provided as zinc chloride alone. These findings are important in that they highlight the importance of both dietary zinc formulation and the use of functional assessments in determining zinc nutriture. PMID- 10827223 TI - Circadian release of hypothalamic norepinephrine in rats in vivo is depressed during early L-lysine deficiency. AB - Rats rapidly recognize an amino acid-deficient diet, presumably via central mechanisms that involve hypothalamic circuits. We evaluated the effects of a deficiency of the essential amino acid, L-lysine, on the ventromedial hypothalamus (VMH) norepinephrine (NE) circadian release in free-moving, nonstressed rats. A dialysis probe was implanted into the VMH of male Wistar rats. Continuous microdialysis measurement was done during the first 26 h of L lysine (Lys) deficiency in rats that had free access to food and fluid. The dark phase was from 1900 to 0700 h. Rats were divided into six groups according to their food and fluid intakes. They were fed either normal (Lys sufficient) or Lys deficient powdered food and provided with distilled water, glycine (Gly, 400 mmol/L) or Lys solution (400 mmol/L). In control rats, VMH NE release showed a diurnal pattern, with the lowest levels measured at the onset of the dark phase. In Lys-deficient rats, the release was significantly depressed from the early morning (0500 h) compared with Lys-sufficient rats, without any differences in food and fluid intakes. A normal pattern of VMH NE was restored by the provision of 400 mmol/L Lys solution to deficient rats. The results suggest that the VMH NE release is involved in the early integration of signals about amino acid deficiency. PMID- 10827224 TI - Skin cancer in liver transplant recipients. AB - Skin cancer is the most common malignancy arising in the posttransplantation setting. Multiple factors contribute to the high risk for cutaneous carcinoma in immunosuppressed organ-transplant recipients. We review the phenomenon of skin cancer in solid-organ transplant recipients and further delineate the problem in the context of liver transplantation. Skin cancer is a significant medical and surgical problem for organ-transplant recipients. With prolonged allograft function and patient survival, the majority of solid-organ transplant recipients will eventually develop skin cancer. Although squamous cell carcinoma is the most common cutaneous malignancy in this population, basal cell carcinoma, melanoma, and Kaposi's sarcoma, as well as uncommon skin malignancies, may occur. Highly susceptible patients may develop hundreds of squamous cell carcinomas, which may be life threatening. Management strategies focus on regular full-skin and nodal examination, aggressive treatment of established malignancies, and prophylactic measures to reduce the risk for additional photodamage and malignant transformation. Skin cancer is a substantial cause of morbidity and even mortality among solid-organ transplant recipients. As a byproduct of immunosuppression, liver transplant recipients experience a high incidence of skin cancer and should be educated and managed accordingly. PMID- 10827225 TI - Liver transplantation for hereditary transthyretin amyloidosis. AB - Transthyretin (TTR) amyloidosis is the most common form of hereditary amyloidosis. It is a systemic amyloidosis caused by an amyloidogenic variant TTR (ATTR), of which the methionine for valine at position 30 (ATTR Val30Met) gives rise to a fatal neuropathic amyloidosis. Because more than 95% of TTR is produced by the liver, a liver transplantation should abolish the liver's production of amyloidogenic mutant TTR and thereby halt amyloid formation. The first liver transplantation for hereditary TTR amyloidosis was performed in Sweden in 1990 on a patient with ATTR Val30Met amyloidosis, and the result was encouraging. Today, liver transplantation for TTR amyloidosis is an established treatment. However, the disease is rarely seen except in a few endemic areas; therefore, most transplantation centers only receive a few cases. Because the disease phenotype varies with different TTR mutations and variability is even encountered for the same mutation, an evaluation of patients for transplantation must include an investigation of all organs that may be affected by the disease and may impact on the morbidity and mortality of the procedure. The aim of this review is to present the results of liver transplantation for TTR amyloidosis and give recommendations for patient evaluation and selection based on the literature and our experience with the disease. PMID- 10827227 TI - New treatments for hepatorenal syndrome. PMID- 10827226 TI - Improvement of hepatorenal syndrome with extracorporeal albumin dialysis MARS: results of a prospective, randomized, controlled clinical trial. AB - In hepatorenal syndrome (HRS), renal insufficiency is often progressive, and the prognosis is extremely poor under standard medical therapy. The molecular adsorbent recirculating system (MARS) is a modified dialysis method using an albumin-containing dialysate that is recirculated and perfused online through charcoal and anion-exchanger columns. MARS enables the selective removal of albumin-bound substances. A prospective controlled trial was performed to determine the effect of MARS treatment on 30-day survival in patients with type I HRS at high risk (bilirubin level, > or =15 mg/dL) compared with standard treatment. Thirteen patients with cirrhosis with type I HRS were included from 1997 to 1999. All were Child's class C, with Child-Turcotte-Pugh scores of 12.4 +/- 1. 0, United Network for Organ Sharing status 2A, and total bilirubin values of 25.7 +/- 14.0 mg/dL. Eight patients were treated with the MARS method in addition to hemodiafiltration (HDF) and standard medical therapy, and 5 patients were in the control group (HDF and standard medical treatment alone). None of these patients underwent liver transplantation or received a transjugular intrahepatic portosystemic shunt or vasopressin analogues during the observation period. In the MARS group, 5.2 +/- 3.6 treatments (range, 1 to 10 treatments) were performed for 6 to 8 hours daily per patient. A significant decrease in bilirubin and creatinine levels (P <.01) and increase in serum sodium level and prothrombin activity (P <.01) were observed in the MARS group. Mortality rates were 100% in the control group at day 7 and 62.5% in the MARS group at day 7 and 75% at day 30, respectively (P <.01). We conclude that the removal of albumin bound substances with the MARS method can contribute to the treatment of type I HRS. PMID- 10827228 TI - Evaluation of 100 patients for living donor liver transplantation. AB - The initial success of living donor liver transplantation (LDLT) in the United States has resulted in a growing interest in this procedure. The impact of LDLT on liver transplantation will depend in part on the proportion of patients considered medically suitable for LDLT and the identification of suitable donors. We report the outcome of our evaluation of the first 100 potential transplant recipients for LDLT at the University of Colorado Health Sciences Center (Denver, CO). All patients considered for LDLT had first been approved for conventional liver transplantation by the Liver Transplant Selection Committee and met the listing criteria of United Network for Organ Sharing status 1, 2A, or 2B. Once listed, those patients deemed suitable for LDLT were given the option to consider LDLT and approach potential donors. Donors were evaluated with a preliminary screening questionnaire, followed by formal evaluation. Of the 100 potential transplant recipients evaluated, 51 were initially rejected based on recipient characteristics that included imminent cadaveric transplantation (8 patients), refusal of evaluation (4 patients), lack of financial approval (6 patients), and medical, psychosocial, or surgical problems (33 patients). Of the remaining 49 patients, considered ideal candidates for LDLT, 24 patients were unable to identify a suitable donor for evaluation. Twenty-six donors were evaluated for the remaining 25 potential transplant recipients. Eleven donors were rejected: 9 donors for medical reasons and 2 donors who refused donation after being medically approved. The remaining 15 donor-recipient pairs underwent LDLT. Using our criteria for the selection of recipients and donors for LDLT gave the following results: (1) 51 of 100 potential transplant recipients (51%) were rejected for recipient issues, (2) only 15 of the remaining 49 potential transplant recipients (30%) were able to identify an acceptable donor, and (3) 15 of 100 potential living donor transplant recipients (15%) were able to identify a suitable donor and undergo LDLT. Recipient characteristics and donor availability may limit the widespread use of LDLT. However, careful application of LDLT to patients at greatest risk for dying on the waiting list may significantly reduce waiting list mortality. PMID- 10827229 TI - Single-center analysis of the first 40 adult-to-adult living donor liver transplants using the right lobe. AB - The first adult-to-adult living donor liver transplant using the right hepatic lobe in the United States was performed only 2 years ago. Although initial reports were encouraging, continuous review of the results and appropriate modifications in patient management will be necessary to minimize donor risk and optimize recipient outcome. The results of 40 such transplantations were analyzed and are summarized. Recipients were listed for transplantation according to the usual criteria. Living donors were not considered for United Network for Organ Sharing status IIA patients after the initial 22 patients. Donor evaluation followed a rigid protocol. A graft-to-recipient body weight ratio of at least 0.8% was the minimum required throughout most of the study. The surgical procedures were similar, except the plane of transection was modified to better accommodate donor biliary anatomy, and uniform stenting of bile ducts was practiced after the first 10 transplants. Immunosuppression consisted of tacrolimus, mycophenolate mofetil, and a prednisone taper. The target tacrolimus level was decreased and mycophenolate was withdrawn more rapidly in the second half of the study because of the absence of acute cellular rejection. Donor morbidity has been limited to minor complications, and transplant recipient biliary complications decreased from 35% to 0%. Acute cellular rejection has not been observed despite less aggressive immunosuppression, and septic complications decreased dramatically. There have been no recipient deaths since these changes were instituted. Right lobectomy can be performed safely in the donor population. Recipient biliary complications can be minimized with stenting. Less aggressive immunosuppression is well tolerated and minimizes septic complications and attributable mortality. PMID- 10827230 TI - Biliary stenting versus bypass surgery for the palliation of malignant distal bile duct obstruction: a meta-analysis. AB - The objective of this analysis is to compare endoscopic stenting with surgical bypass in patients with unresectable, malignant, distal common bile duct obstruction using the technique of meta-analysis. The inclusion criteria for the studies were randomized patient assignment, publication in the English language, 20 or more patients per group, all patients followed up until death, and follow up and complications reported in an equivalent way for both treatment arms. Data extraction was performed independently by 2 of the authors. The number of treatment failures, serious complications, requirement for additional treatment sessions, and 30-day mortality were extracted. Three existing trials met the inclusion criteria, all of which compared surgery with the use of plastic stents. There were no studies identified that used metallic expandable stents. For the rate of treatment failure and serious complications, the odds ratios (ORs) of the 3 trials were heterogeneous, and no summary ORs were calculated. More treatment sessions were required after stent placement than after surgery, and a common OR was estimated to be 7.23 (95% confidence interval [CI], 3.73 to 13.98). Thirty day mortality was not significantly different (OR = 0.522; 95% CI, 0.263 to 1.036). Although surgical bypass required fewer additional treatment sessions, existing data do not allow a definitive conclusion on which treatment is preferable. A larger randomized controlled trial using newer metallic stents and proper quality-of-life instruments is required. PMID- 10827231 TI - Prolonged disease-free survival after orthotopic liver transplantation plus adjuvant chemoirradiation for cholangiocarcinoma. AB - Orthotopic liver transplantation (OLT) alone for unresectable cholangiocarcinoma is often associated with early disease relapse and limited survival. Because of these discouraging results, most programs have abandoned OLT for cholangiocarcinoma. However, a small percentage of patients have achieved prolonged survival after OLT, suggesting that adjuvant approaches could perhaps improve the survival outcome. Based on these concepts, a protocol was developed at the Mayo Clinic using preoperative irradiation and chemotherapy for patients with cholangiocarcinoma. We report our initial results with this pilot experience. Patients with unresectable cholangiocarcinoma above the cystic duct without intrahepatic or extrahepatic metastases were eligible. Patients initially received external-beam irradiation plus bolus fluorouracil (5-FU), followed by brachytherapy with iridium and concomitant protracted venous infusion of 5-FU. 5 FU was then administered continuously through an ambulatory infusion pump until OLT. After irradiation, patients underwent an exploratory laparotomy to exclude metastatic disease. To date, 19 patients have been enrolled onto the study and have been treated with irradiation. Eight patients did not go on to OLT because of the presence of metastasis at the time of exploratory laparotomy (n = 6), subsequent development of malignant ascites (n = 1), or death from intrahepatic biliary sepsis (n = 1). Eleven patients completed the protocol with successful OLT. Except for 1 patient, all had early-stage disease (stages I and II) in the explanted liver. All patients who underwent OLT are alive, 3 patients are at risk at 12 months or less, and the remaining 8 patients have a median follow-up of 44 months (range, 17 to 83 months; 7 of 9 patients > 36 months). Only 1 patient developed tumor relapse. OLT in combination with preoperative irradiation and chemotherapy is associated with prolonged disease-free and overall survival in highly selected patients with early-stage cholangiocarcinoma. PMID- 10827232 TI - Revisiting liver transplantation for patients with hilar cholangiocarcinoma: the Mayo Clinic proposal. PMID- 10827233 TI - Improved survival with screening for hepatocellular carcinoma. AB - Hepatocellular cancer (HCC) is a frequent cause of cancer death worldwide. Although successful screening programs exist in Asian countries, the role for screening in the United States is not clear. The purpose of this study is to determine if screening has an effect on survival in a predominantly Asian population in Hawaii. We retrospectively reviewed cases of HCC from 1993 to 1998 for demographic data, risk factors, symptoms, stage, treatment, and survival. Patients were divided into 3 groups according to presentation: symptomatic (S), asymptomatic (A), and asymptomatic but screened for disease (A/Sc). Kaplan-Meier analysis was performed on overall survival by group. Ninety-one patients were referred for HCC. There were 56 patients in group S, 19 patients in group A, and 16 patients in group A/Sc. Patients in group A/Sc survived significantly longer than those in group S (P =.009), with the former group having a median survival of 1,399 versus 234 days for group S. Median survival for group A was 545 days and did not differ significantly from the other groups. Patients who are asymptomatic and screened for HCC have an increased survival compared with those patients who present with symptoms. This may justify the use of a formal screening program for HCC in Hawaii, but we would need to investigate this prospectively. PMID- 10827234 TI - Early vascular complications after pediatric liver transplantation. AB - Vascular complications have a detrimental effect on the outcome after liver transplantation. Most studies focus exclusively on hepatic artery thrombosis (HAT). The current study analyzed the incidence, consequences, and risk factors for HAT, portal vein thrombosis (PVT), and venous outflow tract obstruction (VOTO) in a consecutive series of 157 pediatric liver transplantations. The overall incidence of vascular complications was 21%. The incidences of HAT, PVT, and VOTO were 10%, 4%, and 6%, respectively. Patient survival after PVT and VOTO and graft survival after HAT and PVT were less compared with survival of grafts without vascular complications. To identify risk factors for vascular complications, factors related to recipient, donor, and surgical techniques were analyzed. A low donor-recipient (D/R) age ratio, long surgical time, and use of the proper hepatic artery of the recipient for arterial reconstruction were risk factors for HAT. Young age, low weight, segmental grafts, and piggyback technique were risk factors for PVT. Fulminant hepatic failure, high D/R age and weight ratios, and use of segmental grafts were related to VOTO. Vascular complications, which occurred in 21% of the pediatric liver transplantations, had a significant impact on patient and graft survival. Size disparity between donor and recipient was an important risk factor for vascular complications, especially in the case of transplantation of segmental grafts. Patient and graft survival might improve by avoiding the identified risk factors. PMID- 10827235 TI - Health-related quality of life in long-term survivors of pediatric liver transplantation. AB - The purpose of this study is to measure the health-related quality of life (HRQOL) in children who are long-term survivors of liver transplantation and to pilot the Liver Transplant Disability Scale (LTDS), a newly developed 12-point scale that quantifies chronic medical disability related to liver transplantation. This study is a cross-sectional survey of 51 children surviving liver transplantation by at least 2 years, with a median age of 4.94 years. Functional capacity and utility scores were measured by the Health Utilities Index Mark II (HUI2), and chronic disease-specific medical disability was measured by the LTDS. HUI2 results were compared with a reference population. LTDS scores were compared with utility scores and patient survival 3 years later. Ninety percent of the study patients had functional deficits compared with 50% of controls. Functional impairment was typically mild. The resulting mean utility score, 0.86 +/- 0.13 (0 = dead, 1 = perfect health), was significantly less than that of the reference population, 0.95 +/- 0. 07 (P <.001). LTDS scores ranged from 0 (no disability) to 6 (moderate disability). Seventy-one percent of the children had mild disability (scores 0 to 3), and 29% had moderate disability (scores 4 to 6). LTDS scores did not correlate with utility scores but were predictive of survival. The majority of pediatric liver transplant recipients have mild functional deficits. Their utility scores reflected a high level of HRQOL but were significantly less than those of a reference population. The majority also had mild medical disability, predominantly delayed growth. Medical disability did not correlate with HRQOL but predicted survival 3 years later. PMID- 10827236 TI - Laparoscopic cholecystectomy and cirrhosis: a case-control study of outcomes. AB - The incidence of gallstone disease in patients with cirrhosis is greater than that in healthy patients. Previous surgical literature reported greater morbidity and mortality in patients with cirrhosis with both open and laparoscopic cholecystectomy (LC). We compared our recent experience with LC in patients with cirrhosis and controls. A retrospective review was performed using the search terms, "cirrhosis" and "laparoscopic cholecystectomy." Forty-eight patients with cirrhosis were identified and randomly matched with healthy controls by age and sex. Four controls were assigned per patient with cirrhosis. Outcomes assessed included mortality, duration of surgery, length of hospital stay, blood transfusion requirement, postoperative complications, and need for conversion to open cholecystectomy. Forty-eight patients with cirrhosis and 187 healthy controls underwent LC. Child-Pugh classification of severity of liver disease was as follows: Child's class A, 38 of 48 patients; Child's class B, 10 of 48 patients; and Child's class C, 0 of 48 patients. Patients with cirrhosis had statistically significantly lower albumin levels (P =.0001) and prolonged prothrombin times (P =. 05). Average duration of surgery for patients with cirrhosis was 1. 71 versus 1.57 hours (P =.57) for controls. Average length of hospital stay for patients with cirrhosis was 6.47 versus 4.77 days (P =.152) for controls. Average number of units of blood transfused in patients with cirrhosis was 0.156 versus 0.0 units (P =.025) in controls. Complications occurred in 6 of 48 patients with cirrhosis (12.5%) and 8 of 187 controls (4.2%; P <.05). No child's class C patient underwent LC. Four patients with cirrhosis (8.3%) and no controls were converted to open cholecystectomy. No postoperative infections were noted. There was no mortality in either group. LC in patients with Child's class A and B cirrhosis is reasonably safe and shows no increase in morbidity or mortality or worsening of outcome. Further studies are required to evaluate the management of acute gallbladder disease in Child's class C patients. PMID- 10827237 TI - Intracerebral hemorrhage after liver transplantation. AB - We report 5 patients with intracerebral hemorrhage after orthotopic liver transplantation (OLT) and identify the possible risk factors. Between November 1991 and April 1999, 75 adult patients received 77 orthotopic liver transplants at Queen Mary Hospital, Hong Kong. Five patients (6.5%) developed intracerebral hemorrhage postoperatively. Clinical and laboratory data were reviewed, and potential risk factors were analyzed. The 5 patients developed intracerebral hemorrhage within 40 days (range, 1 to 37 days; median, 4 days) after OLT. The mortality rate was 80% (4 of 5 patients). The intraoperative blood transfusion volume (median, 17,200 mL; range, 15,750 to 30,360 mL) administered to patients who developed intracerebral hemorrhage postoperatively was significantly greater than that (median, 6,990 mL; range, 1,840 to 22,680 mL) for patients without the complication (P =.0008). Massive intraoperative transfusion (>15,000 mL) was required in all 5 patients (100%) with intracerebral hemorrhage but only 9 of 72 patients (12.5%) in the other group (P =.0001). Four of 5 patients (80%) with intracerebral hemorrhage had intraoperative hypotension compared with 7 of 72 patients (9.7%) in the other group (P =.001). No significant difference was found in age, prothrombin time (PT), activated partial thromboplastin time (APTT), incidence of hypertension, bleeding at extracerebral sites, cyclosporine A neurotoxicity, thrombocytopenia, hemodialysis, and sepsis between the patients with and without intracerebral bleeding. However, the median cumulative score of coagulation parameters (PT, APTT, platelet count) was significantly greater in the group with than without intracerebral bleeding (median score, 3 v 1; P =.023). Intracerebral hemorrhage is 1 of the most disastrous complications after OLT. Intraoperative hypotension, massive intraoperative transfusion, and coagulopathy may be correlated with this complication. PMID- 10827238 TI - Erythropoietin response to post-liver transplantation anemia. AB - Anemia frequently accompanies end-stage liver disease. Erythropoietin has recently been shown to be of benefit in a number of diseases complicated by anemia. We studied erythropoietin levels before and after orthotopic liver transplantation (OLT) and correlated these with the degree of anemia. Twenty seven patients with end-stage cirrhosis who underwent OLT had preoperative and weekly postoperative serum erythropoietin levels determined by a highly sensitive radioimmunoassay. The relation of erythropoietin level to the values for hematocrit, serum creatinine, and cyclosporine and other biochemical test results was evaluated. Before transplantation, 23 patients were anemic; erythropoietin levels were appropriately elevated (72.7 +/- 37 mU/mL; normal, 10 to 15 mU/mL) for the degree of anemia (hematocrit, 33.1% +/- 1%) in 16 patients (70%). A blunted erythropoietin response to anemia was found in 7 of the anemic patients with cirrhosis (30%). After OLT, the hematocrit decreased to 29.5% +/- 0.6% at 4 weeks, with a reciprocal increase in serum erythropoietin levels to 36 +/- 5 mU/mL. Erythropoietin response appeared appropriate to the degree of anemia in 82% of the liver transplant recipients and blunted in 18%. We conclude that the ability to secrete erythropoietin in response to anemia is defective in many patients with end-stage liver disease, and a normal response may be restored after OLT. The results suggest that exogenous erythropoietin administration may be beneficial in anemic patients with cirrhosis and liver transplant recipients who have inappropriately low serum erythropoietin levels. PMID- 10827239 TI - Internal biliary stenting in orthotopic liver transplantation. AB - Biliary complications account for significant morbidity in orthotopic liver transplantation (OLT), with a reported incidence ranging from 6% to 47%, and many centers are reassessing the need and options available for stenting the biliary anastomosis. We report on our experience using a 6F Silastic, double-J, ureteral stent as an internal biliary stent in OLT. From October 15, 1995, to September 30, 1998, a total of 99 patients at our institution underwent 108 OLTs. Of these, 77 patients received an end-to-end choledochocholedochostomy over an internal stent. Three patients died within 1 week post-OLT, leaving 74 patients for evaluation (follow-up, 2 to 38 months). Stents were placed transanastomotic and transsphincteric at the time of OLT and secured with a dissolvable suture. At 4 to 6 weeks post-OLT, stents visible within the biliary tree on kidney, ureters, and bladder radiograph were removed endoscopically. Graft and patient survival rates were 92% and 96%, respectively. There were 12 biliary complications (18%): anastomotic leak in 6 patients (9%), anastomotic stricture in 5 patients (7.6%), and stent migration in 1 patient (1.5%). Thirty-two patients (43%) passed the biliary stent without intervention, whereas 42 patients (57%) underwent esophagogastro duodenoscopy (EGD) stent removal at 4 to 6 weeks without incident. Treatment of the complications included percutaneous drainage, endoscopic dilatation with stenting, and/or conversion to Roux-en-Y choledochojejunostomy. The use of the 6 F Silastic, double-J, ureteral stent provides a safe and effective means of stenting the biliary anastomosis in OLT. Major advantages to this method are that it: (1) is completely internal, (2) is biliary decompressive, (3) is radiopaque, (4) can be spontaneously passed, and (5) is easily accessible for EGD extraction. PMID- 10827240 TI - Pancreatitis in adult orthotopic liver allograft recipients: risk factors and outcome. AB - Acute pancreatitis (AP) has been described after orthotopic liver transplantation but is uncommon in stable patients after the initial perioperative phase. The aim of this study is to review our experience with AP occurring more than 2 months after primary allografting and determine possible contributing factors plus patient outcome. A review of patient files and the unit database was performed. AP was diagnosed in 9 of 298 patients (3%) on 12 occasions. The incidence of AP was greater in men (8 of 163 men) than women (1 of 135 women; P <.04). Underlying factors to each episode of AP were biliary manipulation (4 of 12 episodes; 33%), history of recent alcohol ingestion (3 of 12 episodes; 25%), and malignancy in the region of the pancreas (2 of 12 episodes; 16%). AP was associated with a diagnosis of either hepatic artery thrombosis combined with biliary tract complications (P <.005) or malignancy (P <.004). In 7 of 12 episodes of AP (58%), conservative management alone was successful. In 3 of 9 patients (33%), subsequent surgery was required. One patient died of pancreatic malignancy. In conclusion, AP is uncommon in stable liver transplant recipients. Male sex, complications of hepatic artery thrombosis, and malignancy in the region of the pancreas are associated with AP in this study. PMID- 10827241 TI - Hepatic arterial anatomy as applied to living-donor and split-liver transplantation. PMID- 10827242 TI - Reversibility of alcohol-related brain atrophy. PMID- 10827243 TI - Alcohol and the brain: a love and hate relationship. PMID- 10827244 TI - Delayed recurrence of hepatocellular carcinoma after liver transplantation: detection of origin by chromosomal analysis. AB - We report on a 41-year-old man undergoing liver transplantation for chronic hepatitis C who presented 26 months later with hepatocellular carcinoma. No evidence of hepatocellular carcinoma could be found in the native liver, although features of small cell dysplasia were prominent. Although he had recurrent hepatitis C, the transplanted liver was not cirrhotic. Chromosomal analysis was used to resolve whether this was a de novo tumor or a recurrence of an unsuspected tumor present at the time of transplantation. This male patient had received a liver from a female donor, and in situ hybridization for the Y chromosome showed reactivity in the tumor but not in surrounding nontumorous liver. Thus, this is an example of the use of chromosomal analysis to resolve the origin of a tumor occurring in the transplant setting. PMID- 10827245 TI - Prophylactic fluconazole in liver transplant recipients: a randomized, double blind, placebo-controlled trial. PMID- 10827246 TI - Analytic modeling to predict the outcome of chronic hepatitis C infection: a useful clinical tool or an unfair representation of natural history? PMID- 10827247 TI - Tumor necrosis factor gene promoter polymorphism and recurrent hepatitis C after liver transplantation: the missing link to pathogenesis or a casual association? PMID- 10827249 TI - Call for papers PMID- 10827248 TI - Xenotransplantation: a summary of the International Business Communications' Fourth International Congress. PMID- 10827250 TI - Longitudinal monitoring of WBC subsets in packed RBC units after filtration: implications for transfusion transmission of infections. AB - BACKGROUND: Specific subsets of peripheral blood WBCs are reservoirs for infectious agents, such as CMV and EBV, and can serve as vectors for transfusion transmission of these agents. While filter WBC reduction has been used to prevent transfusion transmission of infections, its effectiveness has not been documented for many infectious agents and in some instances may be difficult to demonstrate in clinical trials. Because the effectiveness of filtration depends on the number of infected WBCs remaining at transfusion, WBC subpopulations in packed RBC units were quantitated after filtration and storage. STUDY DESIGN AND METHODS: Packed RBC units (n = 14) were filtered and stored at 4(o)C for 42 days or were stored without filtration. Serial samples were subjected to flow cytometric immunophenotyping of WBC subsets: neutrophils, monocytes, CD4+ and CD8+ T cells, B cells, and NK cells. RESULTS: Filtration produced a mean reduction in total WBCs of 3.2 log. Monocytes, lymphocytes, and neutrophils were reduced by 4.1, 3.8, and 2.5 log, respectively. Lymphocyte subsets also demonstrated differential reduction with filtration. All WBC subsets showed ongoing loss during storage. CONCLUSIONS: Monocyte and lymphocyte subsets are removed most effectively by prestorage filtration. Postfiltration storage leads to further significant reductions in WBC subsets. The implications of these findings for the mitigation of transfusion transmission of infection are discussed. PMID- 10827251 TI - Immunomagnetic selection of CD34+ cells: factors influencing component purity and yield. AB - BACKGROUND: In immunomagnetic selection of CD34+ cells from HPC transplants, not all factors that affect yield and purity of CD34+ cells are known. METHODS: Forty three consecutive procedures of immunomagnetic selection of CD34+ cells from peripheral blood HPCs and bone marrow harvests (autologous harvests, n = 27; allogeneic harvests; n=16) were performed by use of a cell selection system (Isolex 300i, Baxter Immunotherapy). The composition of the starting component and the subsets of CD34+ cells were analyzed for correlation with the yield and purity of the final component. RESULTS: The mean purity of the final components was 84.3 percent (range, 27-99%), and the mean yield was 51.4 percent (range, 9.4 80. 4%). Partial regression analysis showed that, among the factors correlating with purity and/or yield, the RBC volume in the starting fraction had the highest predictive impact on the purity and yield of CD34+ cells, even after the exclusion of procedures using bone marrow harvests as an HPC source (beta coefficient, -0.704; p = 0. 001). CONCLUSION: The use of the Isolex 300i system allows efficient recovery of CD34+ cells in routine selection procedures. The volume of RBCs in the starting component should be minimized to ensure a high yield and purity of the final component. PMID- 10827252 TI - A multicenter study evaluating three methods for counting residual WBCs in WBC reduced blood components: Nageotte hemocytometry, flow cytometry, and microfluorometry. AB - BACKGROUND: A multicenter study was conducted to evaluate the performance characteristics of flow cytometry and microfluorimetry for counting low concentrations of WBCs and to compare the results with Nageotte hemocytometry. STUDY DESIGN AND METHODS: A two-phase study involving 10 centers located in the United States and in Europe was performed. Coded samples of RBCs and platelets were distributed by 24-hour (Phase 1) or 2-day (Phase 2) courier service to each test site for analysis. Samples were prepared to include concentrations of WBCs slightly above and below the concentration corresponding to the threshold standards for WBC-reduced RBCs and platelets. All centers tested samples by Nageotte hemocytometry plus one or both of two automated methods. RESULTS: Both flow cytometry and microfluorometry gave better results than Nageotte hemocytometry in testing freshly prepared samples. At WBC concentrations >5 per microL (RBCs) or >3 per microL (platelets), the intersite CV was <20 percent for the automated methods but >30 percent for the Nageotte hemocytometer method (p<0.001). Accuracy was greater for the automated methods than for the Nageotte hemocytometer method (p<0. 001). Nageotte hemocytometry showed a bias to underestimation relative to the results obtained with the automated methods. All methods had poorer performance in testing samples that required > or =2 days' shipment than in testing of those requiring overnight shipment. CONCLUSION: Automated methods for counting residual donor WBCs in WBC-reduced cellular components offer advantages of improved precision and greater accuracy than are seen with the Nageotte hemocytometer method. Automated methods are less labor intensive but more costly than microscopic methods. Preparation and shipping methods will need further refinement for samples to be counted more than 24 hours after sample collection. PMID- 10827253 TI - Filtration of RBC units:effect of storage time and temperature on filter performance. AB - BACKGROUND: The influence of time, temperature, and rate of filtration on the efficacy of WBC reduction of RBC units was studied in a controlled, paired-donor format. STUDY DESIGN AND METHODS: Ten donors underwent whole-blood phlebotomy on two to four occasions each. Units were filtered (RCXL-1, Pall Biomedical) under laboratory conditions and gravity flow as follows: 1) after 0 to 2 hours of storage at 22 degrees C, 2) after 7 to 8 hours at 22 degrees C, 3) after 14 days of storage at 4 degrees C, and 4) under mock bedside conditions after 14 days of storage at 4 degrees C. Prefiltration and postfiltration cell counts and prefiltration WBC CD11a expression were assessed on Days 0 and 14. RESULTS: WBC content before filtration was 2.20 and 2.34 x 10(9) (p>0.05) for units stored for 2 and 8 hours (Groups 1 and 2) and declined to 52.8 and 7. 57 x 10(4) (p<0.01) after filtration. The efficacy of WBC reduction in units stored for 14 days was similar to that in units stored for 8 hours, but absolute postfiltration WBC counts were significantly lower because of a 0.6 log reduction in the starting WBC count after 14 days of storage (postfiltration WBC content of 1.02 and 2.31 x 10(4) for units filtered under laboratory vs. bedside conditions [p>0.05]). Filtration under bedside conditions was associated with a greater degree of variation in residual WBC counts than laboratory filtration. WBC reduction by filtration was significantly greater in units stored for at least 8 hours (Groups 2, 3, and 4) than in those stored for less than 2 hours (4.59 log vs. 3.83 log reduction in WBC content, p<0.05). Surface expression of leukocyte function antigen 1 as measured by CD11a was similar in all groups. CONCLUSION: WBC reduction of RBC units by filtration was least effective when performed within 2 hours of collection. Efficacy of WBC reduction increased significantly after the units were stored for 8 hours to 14 days, without significant differences between these storage intervals. Laboratory filtration yielded more consistent results than did mock bedside filtration. Temperature and filtration rate had no effect on the efficacy of WBC reduction by filtration. PMID- 10827254 TI - Effects of the addition of second-messenger effectors to platelet concentrates separated from whole-blood donations and stored at 4 degrees C or -80 degrees C. AB - BACKGROUND: Platelet concentrates (PCs) are currently stored at 22 degrees C under continuous agitation. Because of the potential risk of the overgrowth of bacteria in case of contamination, PC shelf life is limited to 5 days. A mixture of second-messenger effectors is being evaluated to determine if it has benefits for cold liquid storage and cryopreservation of platelets. STUDY DESIGN AND METHODS: PCs separated from whole-blood donations by the buffy coat method were randomly assigned (n = 6 each) to be stored for 5 days at 22 degrees C under continuous agitation or at 4 degrees C after treatment with a platelet storage medium (ThromboSol, LifeCell Corp. ). PCs were also cryopreserved with 6-percent DMSO (final concentration) or with ThromboSol plus 2-percent DMSO (final concentration) (TC). After storage, platelets were analyzed by flow cytometry, transmission electron microscopy, and aggregation and perfusion techniques. RESULTS: Cold liquid storage of ThromboSol-treated platelets resulted in a lower binding of coagulation factor Va on the platelet surface than on platelets stored at 22 degrees C. In transmission electron microscopy, a conversion to spherical morphology was seen in the case of cold liquid storage. No difference between ThromboSol-treated platelets stored at 4 degrees C and platelets stored at 22 degrees C was seen in perfusion studies. Cryopreservation in the presence of TC prevented the reduction in glycoprotein Ib and IV expression on platelet surface that is seen in 6-percent DMSO-cryopreserved platelets. Platelets cryopreserved in TC covered, by thrombus, a significantly greater percentage of the perfused surface after the freezing and thawing process. CONCLUSION: ThromboSol-treated PCs separated from whole-blood donations by the buffy coat method, stored at 4 degrees C for 5 days, or cryopreserved in the presence of TC, maintained in vitro functional activity comparable to that achieved by current methods of storage, although discoid morphology was not preserved during cold liquid storage with ThromboSol. PMID- 10827255 TI - Alterations in cytoskeletal organization and tyrosine phosphorylation in platelet concentrates prepared by the buffy coat method. AB - BACKGROUND: Numerous morphologic and biochemical changes occurring during platelet storage may result in the impairment of platelet function. STUDY DESIGN AND METHODS: The effect of preparation and storage conditions on platelet function was analyzed through evaluation of cytoskeletal organization and signaling mechanisms involved in the activation of platelets by thrombin. Samples of platelets prepared by the buffy coat method were obtained before and after the platelet concentrates were prepared during storage for 1, 3, and 5 days. Thrombin induced aggregation was monitored, and changes in the organization of proteins in the cytoskeleton were analyzed by gel electrophoresis. For the analysis of tyrosine phosphorylation, proteins were transferred to nitrocellulose membranes and probed with a specific antibody. RESULTS: The aggregation and the cytoskeletal organization induced by thrombin activation were markedly impaired immediately after preparation of platelet concentrates, although they normalized after the first 24 hours of storage and decreased progressively after 3 days of storage. Results in tyrosine phosphorylation paralleled those obtained with cytoskeletal organization, except for samples obtained immediately after processing to obtain platelet concentrates. CONCLUSION: These data indirectly suggest that the stress induced by the preparation method has an activating effect on platelet function that may imply a delayed platelet response to further stimuli. This effect may result in a deficient redistribution of signaling molecules within platelets. PMID- 10827256 TI - Regeneration of erythropoiesis after related- and unrelated-donor BMT or peripheral blood HPC transplantation: a major ABO mismatch means problems. AB - BACKGROUND: Blood group incompatibility in allogeneic BMT is common but does not appear to affect the outcome in terms of incidence of graft rejection or delayed engraftment. However, major ABO incompatibility may be associated with prolonged erythroid aplasia. STUDY DESIGN AND METHODS: In a retrospective analysis of 286 allogeneic transplant recipients, the prevalence of prolonged erythroid aplasia, including pure RBC aplasia, was determined. RESULTS: Patients receiving major ABO incompatible grafts showed a significant delay in reticulocyte engraftment (median, 32 days; range, 12-347) from that in patients receiving ABO-identical (20; 10-152) or minor ABO-incompatible (21; 12-47) grafts. Pure RBC aplasia occurred in 7 (3%) of 240 evaluable recipients and was observed only in the major ABO-incompatible group (7/43, 16%). Treatment of pure RBC aplasia consisted of either plasma exchange, which resulted in a response within 16 to 68 days, or immunoadsorption, in which the response occurred between Days 119 and 204 after initiation of treatment. CONCLUSION: Major ABO incompatibility may lead to delayed reticulocyte engraftment, resulting in prolonged transfusion dependency and increased risks of transmission of infection and iron overload. Therefore, therapeutic strategies should be taken into consideration to allow erythroid reconstitution in these patients. PMID- 10827257 TI - Detection of multiple passively acquired alloantibodies following infusions of IV Rh immune globulin. AB - BACKGROUND: Passively acquired blood group alloantibodies are detected regularly after infusions of IV Rh immune globulin (RhIG) for the treatment of immune thrombocytopenic purpura (ITP) in D+ patients. STUDY DESIGN AND METHODS: Blood samples from 16 D+ patients with ITP were tested after treatment with IV RhIG for the presence of passively acquired alloantibodies. Similar studies were conducted for three D- patients after injections of IM RhIG for Rh immunoprophyl-axis. Four production lots of IV RhIG and 2 lots of IM RhIG were tested for the presence of alloantibodies. RESULTS: All 16 D+ patients with ITP developed a positive DAT, as well as positive antibody detection test results, after infusions of IV RhIG. All postinfusion plasma samples contained anti-D, as well as one or more additional antibodies, usually anti-C, -E, -G, -V, or -Fy(a). Eluates from patients' RBCs with positive DAT results contained multiple passively acquired alloantibodies. Multiple alloantibodies were detected in samples of different production lots of IV RhIG or IM RhIG. No acute transfusion reactions were observed in five D+ patients with ITP who had been treated with IV RhIG and had been given serologically incompatible D+ RBCs. After injections of IM RhIG, the only passively acquired alloantibody detected was anti-D. CONCLUSION: Plasma samples from D+ patients with ITP treated with IV RhIG regularly contained anti-D and multiple other passively acquired Rh, Duffy, or Kidd system alloantibodies. Postinfusion RBC samples all had positive DAT results with eluates containing anti-D and multiple other Rh, Duffy, or Kidd system antibodies. The consistent detection of multiple passively acquired alloantibodies after IV RhIG, in contrast to the detection of anti-D only after IM RhIG, reflects the immediate effect of the entire (bolus) dose of RhIG by the IV route, the dose for treating ITP that is approximately 10 times the dose for Rh immunoprophylaxis, and the expected serologic incompatibility with recipients' D+ RBCs. PMID- 10827258 TI - The low-frequency MNS blood group antigens Ny(a) (MNS18) and Os(a) (MNS38) are associated with GPA amino acid substitutions. AB - BACKGROUND: Antigens of the MNS blood group system are located on two sialoglycoproteins, GPA and GPB, encoded by GYPA and GYPB. The molecular backgrounds of the low-frequency antigens Ny(a) and Os(a) are not known. STUDY DESIGN AND METHODS: Immunoblotting and a monoclonal antibody-specific immobilization of erythrocyte antigens (MAIEA) assay were used to analyze Os(a). PCR-amplified products of the coding exons of GYPA were studied by single-strand conformation polymorphism analysis, and exon 3 was sequenced. Synthetic peptides were used in hemagglutination-inhibition tests. RESULTS: Sequencing of GYPA exon 3 of two unrelated Ny(a+) persons revealed heterozygosity for a T194A base change encoding an Asp27Glu substitution. Immunoblotting with anti-Os(a) and an MAIEA assay with MoAbs to GPA showed that Os(a) is on GPA. Sequencing exon 3 of an Os(a+) person from the only family with Os(a) revealed heterozygosity for a C273T base change encoding a Pro54Ser substitution. A synthetic peptide representing part of GPA with the Os(a) mutation (VRTVYPSEEETGE) completely inhibited anti Os(a), whereas the control peptide (VRTVYPPEEETGE) did not inhibit anti-Os(a). CONCLUSION: Ny(a) and Os(a) are low-frequency antigens of the MNS blood group system that represent Asp27Glu and Pro54Ser substitutions in GPA, respectively. PMID- 10827259 TI - Identification of a novel hybrid glycophorin gene encoding GP.Hop. AB - BACKGROUND: The GP.Hop (Mi.IV) phenotype expresses the MNS low-incidence antigens Mur, Hop, TSEN, MINY, and MUT. Because serologically similar MNS phenotypes expressing some or all of these antigens were shown to be carried by hybrid GP(B A-B) proteins, it was proposed that a similar protein would be found for GP.Hop. The identification of a second GP.Hop propositus (ES) initiated a study to determine the molecular basis of this phenotype. STUDY DESIGN AND METHODS: Serologic tests and immunoblotting analysis with glycophorin-specific antibodies were performed. GYPB, the gene encoding the GPB protein, was cloned and sequenced after reverse transcription PCR amplification of total RNA isolated from ES. GYPB specific primers encompassing GYPB pseudoexon 3, intron 3, and exon 4 were also used to clone and sequence genomic DNA from ES and MH, the original GP.Hop proband. RESULTS: Serologic and immunochemical data confirmed that ES's RBCs carried antigens associated with the GP.Hop phenotype. Sequencing of ES's cDNA demonstrated the presence of genes predicted to encode s-specific GPB and an S specific GP(B-A-B) hybrid in which the 3' end of GYPB pseudoexon 3 had been replaced by a short nucleotide sequence from exon 3 of the GPA gene (GYPA). The hybrid nucleotide sequence contained sequence motifs previously shown to be required for the expression of the Mur, Hop, TSEN, MINY, and MUT, which is consistent with their presence as detected serologically. Genomic DNA analysis found that the crossover point in GYPB pseudoexon 3 was identical in ES and MH. CONCLUSIONS: The GP.Hop phenotype is produced by a hybrid GP(B-A-B) protein caused by a DNA insertion of GYPA into GYPB. The composition of the hybrid protein is GPB(1-26)-GPpsiB(27-50)-GPA(51-58)-GPB(S)(59-103). PMID- 10827260 TI - Immune-mediated agglutination of cytoskeleton-free RBC microvesicles. AB - BACKGROUND: The factors contributing to RBC agglutination are complex. The RBC cytoskeleton's participation in and contribution to this phenomenon are difficult to separate from those of the plasma membrane. Immunoreactive, cytoskeleton-free, band 3-enriched microvesicles can be generated from normal RBCs. Band 3 has been defined as an important antigen in autoimmune hemolytic anemia (AIHA). STUDY DESIGN AND METHODS: RBC microvesicles devoid of major cytoskeletal proteins were generated and sensitized with eluates obtained from AIHA patients, DAT-positive blood donors, and antisera to common RBC antigens. Monoclonal anti-human IgG was added and agglutination was investigated. Autoantibody-specific binding was evaluated by employing (125)I protein A. RESULTS: RBC vesicle agglutination with a 4+ anti-human globulin score was obtained with 10 autoantibody eluates from AIHA patients and anti-D (3+), but not with eluates from 20 DAT-positive blood donors or antisera directed to eight other common RBC antigens. Microvesicles sensitized with AIHA eluates bound 67 to 167 times as much (125)I protein A radioactivity as did those incubated with buffered normal saline and 18 to 45 times more than vesicles incubated with normal serum. CONCLUSION: The major proteins of the RBC cytoskeleton are not required for supporting IgG immune mediated agglutination of RBC microvesicles. PMID- 10827262 TI - Evaluation of automated RNA-extraction technology and a qualitative HCV assay for sensitivity and detection of HCV RNA in pool-screening systems. AB - BACKGROUND: The objective of this study was the evaluation of NAT technology for the detection of HCV RNA in plasma pools according to the recommendations of the Paul Ehrlich Institute (5000 IU/mL/donation) and the Committee for Proprietary Medical Products (100 IU/mL/manufacturing pool). STUDY DESIGN AND METHODS: Serial dilutions of both the EUROHEP standard (3,800 genome equivalents [geq]/mL; HCV genotype 1) and the World Health Organization (WHO) international standard (100,000 IU/mL; HCV genotype 1) were made in S/D plasma (ESPEP plasma, OctaPharma), which was nonreactive in serologic tests. Serial dilutions of plasma (2 mL) were used for extraction of HCV RNA with an automated version of a nucleic acid isolation method (NucliSens Extractor, Organon Teknika). HCV RNA was co extracted from 2 mL of plasma, together with 84 copies of an in vitro-synthesized single-strand RNA serving as internal extraction control (IC) to monitor the efficiency of extraction and PCR. Amplification and detection of both HCV RNA and IC RNA were performed with an automated PCR system and a qualitative HCV assay (COBAS Amplicor 2.0 HCV, Roche Diagnostics). RESULTS: A cutoff value of 16 geq per mL (10/10 runs [100% hit rate]) was found by using the EUROHEP standard, whereas the WHO international standard had a cutoff value of approximately 12 IU per mL (10/10 runs [100% hit rate]). The IC had a cutoff value of approximately 17.5 copies per mL (6/6 runs [100% hit rate]). Forty-two copies per mL of IC RNA were found in 282 of 284 runs (99% hit rate). The negative controls (ESDEP plasma) were negative in all experiments. Experiments with pool sizes of 12, 24, 48, and 96 using serial dilutions of the WHO international standard revealed a cutoff value of 8 IU per mL (100% hit rate). The EUROHEP standard and the WHO international standard were detected with a 50 percent detection endpoint of 5.2 geq per mL and 1.5 IU per mL, respectively. CONCLUSION: This test system (NucliSens Extractor, and the COBAS Amplicor 2.0 HCV assay) revealed a high sensitivity for HCV RNA; considering the proposed requirements for sensitivity of NAT assays for the detection of HCV RNA in donor plasma, pool sizes of about 400 donors are possible. These endpoint results indicated that 1 IU is equal to about 3.4 geq. PMID- 10827261 TI - Severe HDN due to anti-Ce that required exchange tranfusion. AB - BACKGROUND: Rh system antibodies are commonly encountered in blood bank practice as well as during pregnancy. Nevertheless, no examples of anti-Ce (RH7) have been reported as a cause of HDN that requires exchange transfusion. CASE REPORT: A 38 year-old woman in her fourth pregnancy was typed as blood group O D+, C-, c+, E+, e-. Anti-C and anti-e were detected in her serum during a routine prenatal work up. Further evaluation, including flow cytometric analysis, revealed the presence of a strong anti-Ce and a weak anti-e. Her partner was typed as group A D+, C+, c , E-, e+. A seemingly healthy male infant was delivered at 40 weeks of gestation. The infant's RBCs were typed as group O D-, C+, c+, E+, e+ with a positive DAT (titer 128). Twenty-five hours after birth, the baby had to be transferred to the neonatal intensive care unit because of rapidly rising total serum bilirubin. Despite intensive treatment, including double phototherapy, albumin infusion, and the administration of furosemide and IVIG, the total serum bilirubin level increased during the following day and exchange transfusion with 2 units of type O D-, C-, c+, E+, e- had to be performed; this resulted in a prompt decrease in total serum bilirubin without relapse. CONCLUSION: Anti-Ce caused severe HDN requiring exchange transfusion. This highlights the need for a close follow-up throughout pregnancy if unexpected RBC antibodies are present, to permit the provision of compatible blood in case of a rare antibody. PMID- 10827263 TI - Results of HCV screening of volunteer blood donors with a chemiluminescent immunoassay and a second- or third-generation EIA: overlap of false-positive reactivity and its impact on donor management. AB - BACKGROUND: This study reports the results of adopting a strategy of anti-HCV testing of volunteer blood donors that uses a primary screening assay, two secondary EIAs (Anti-HCV Version III, Murex; Monolisa Anti-HCV New Antigens, Sanofi Pasteur), and a confirmatory immunoblot (HCV WB, Murex). STUDY DESIGN AND METHODS: A comparison was made of HCV test results from volunteer donors tested in two periods when different primary HCV screening assays were in use. The same two secondary screening assays and the same confirmatory test were used for the whole study. The two different primary assays were semi-automated second- or third-generation HCV EIA (Abbott Diagnostics) and an HCV chemiluminescent immunoassay (ChLIA), performed on a fully automated analyzer (PRISM, Abbott). RESULTS: During the period of use of the EIAs as primary screening assays, there were 60 donors per year who were confirmed as anti-HCV-positive, 29 who were classed as having indeterminate HCV serologic results, and 236 who were assessed as having biologically false-positive anti-HCV results. These numbers compared with 57, 52, and 320 such donors, respectively, in the first year of routine use of the ChLIA. The significant increase (p<0.05) in the number of anti-HCV indeterminate donors after the introduction of the ChLIA was primarily due to an increase in donors who reacted on Monolisa HCV, but not an HCV Murex (expected 18/year vs. the observed 31/year, p<0.01). CONCLUSIONS: Compared to the second- or third-generation HCV EIA, the HCV ChLIA has a significantly greater overlap of false reactivity with the Monolisa HCV assay. This finding has implications for the selection of primary and secondary assays for anti-HCV screening of blood donors. PMID- 10827264 TI - False-negative testing errors in routine viral marker screening of blood donors. For the Retrovirus Epidemiology Donor Study. AB - BACKGROUND: The contribution of testing errors to the risk of virus transmission by transfusion depends on the rate of false-negative testing errors and the prevalence of infected seropositive donations. Although the false-negative testing error rate has been estimated at 0.1 to 1 percent on the basis of proficiency studies, it has not previously been measured in routine donor screening. STUDY DESIGN AND METHODS: A 1991 to 1995 database containing 5,153,153 donations from 1.5 x 10(6) donors (including autologous donors) was searched to identify donors who tested seropositive for HIV, HCV, HTLV-I or II and who attempted subsequent donations. The false-negative rate in routinely screened follow-up donations was determined, and false-negative cases were investigated to identify the cause. RESULTS: Subsequent donations (n = 2015) by 1224 donors with confirmed-positive results were identified. Eleven (0.5%) of these donations did not react in EIA. Ten of the 11 false-negative cases were attributable to borderline-reactive donations. On subsequent donations, there were borderline nonreactive results on HTLV-I (2 cases), first-generation HCV (5 cases), and second-generation HCV (3 cases) EIAs. The final case was strongly reactive for HCV in a second-generation EIA on two donations (signal-to-cutoff [S:C] ratio >3.5), followed by a baseline nonreactive result on a third donation (S:C = 0.05). CONCLUSION: False-negative testing results occur infrequently during routine infectious-disease donor screening. Although most false-negative results occurred with borderline-reactive HTLV-II samples and/or early-generation HCV EIAs, frank technical errors (e.g., sample mixup or failure to add sample to EIA well) also occur at a low rate (0.05%; 95% CI, 0-1.5%). Process enhancements designed to reduce errors (e.g., enhanced automation of data management and testing systems and process controls for EIAs) are warranted to detect and prevent false-negative results. PMID- 10827265 TI - High prevalence of TT virus infection in French blood donors revealed by the use of three PCR systems. AB - BACKGROUND: The purpose of this study was to determine the prevalence of TT virus (TTV) infection in voluntary blood donors in Southeastern France. STUDY DESIGN AND METHODS: The sera of 289 blood donors were tested for the presence of TTV DNA by two PCR systems detecting genes located in the 5' UTR (primer set A [Set A]) and the open reading frame (ORF2) (primer set B [Set B]) of the viral genome. A randomized sample of 40 blood donors was also tested by a nested-PCR system in the ORF1 by use of primer set C (Set C). Donors were questioned for possible risk factors for virus transmission. RESULTS: In the entire population studied, 30.8 percent of blood donors tested positive with both Sets A and B, and 70.6 percent with at least one set. In the sample tested with three sets of primers, 27.5 percent of blood donors were positive in testing with all PCR systems and 80 percent with at least one system. The specificity of TTV DNA amplification was confirmed by sequencing 10 PCR products obtained with each set of primers. Statistical analysis revealed that the prevalence of TTV reactivity increased with age. CONCLUSION: The high prevalence of TTV reactivity and the absence of a pathologic condition or risk factors obviously associated with the infection in blood donors suggest that there is no need for systematic detection of TTV infection before blood donation. Further studies are required to determine if TTV isolates can be responsible for a pathologic condition in humans after blood transfusion. PMID- 10827266 TI - Incidence and clinical presentation of posttransfusion TT virus infection in prospectively followed transfusion recipients: emphasis on its relevance to hepatitis. AB - BACKGROUND: A novel transfusion-transmissible human DNA virus, TT virus (TTV), has been discovered recently. An attempt was made to determine the incidence and clinical outcome of TTV infection in recipients of blood transfusion. STUDY DESIGN AND METHODS: Serial serum samples collected as part of a prospective study of posttransfusion hepatitis were examined for TTV DNA by a nested PCR assay. RESULTS: Among 150 adults undergoing cardiac surgery, posttransfusion specimens from 59 individuals were positive for TTV DNA. Pretransfusion sera were found to be positive in 13 of these individuals. Therefore, 46 (33.6%) of the 137 previously uninfected patients developed new TTV viremia after transfusion. Among the 46 patients, 3 were coinfected with HCV, 5 were coinfected with HGV, and 38 were infected with TTV alone. No apparent symptoms or signs were noted in the 38 patients infected by TTV alone or the 5 infected with HGV plus TTV. The average peak serum ALT activity was 31 IU per L, with persistently normal levels in 34 of the 38 patients with TTV infection alone. In 8 other patients who subsequently developed well-documented non-A-G hepatitis, 3 were positive for TTV (3/8 vs. 46/137, p = 0.8). In 12 patients followed for more than 1 year, TTV viremia persisted in every case. CONCLUSION: In this population, TTV is transmitted by transfusion to approximately 30 percent of patients who undergo cardiac surgery. Most of the infections appear to become persistent. Despite the high prevalence rate, TTV does not appear to cause hepatitis on its own. PMID- 10827267 TI - High rate of GB virus type C/HGV transmission from mother to infant: possible implications for the prevalence of infection in blood donors. AB - BACKGROUND: Because GB virus type C(GBV-C)/HGV (GBV-C/HGV) is blood-borne and sexually transmitted, persons at risk of infection with such viruses have a high prevalence of GBV-C/HGV markers. However, adults with no apparent risk factors, such as blood donors, frequently are positive for GBV-C/HGV markers. Mother-to infant transmission could explain this high prevalence, but it has been studied only through small series of GBV-C/HGV-infected mothers co-infected with HCV or HIV. STUDY DESIGN AND METHODS: To determine the rate of mother-to-infant transmission of GBV-C/HGV RNA in women who are HCV- or HIV-negative, a prospective study was performed in a cohort of 288 mothers screened for viral RNA and in the infants born to GBV-C/HGV-infected mothers. RESULTS: Thirteen mothers (4.5%) were found positive for GBV-C/HGV RNA. Of the infants in whom at least one blood sample was collected between the third and the ninth months of life, 89 percent were positive for viral RNA. The majority of these newborns were negative for GBV-C/HGV RNA at birth and positive after the third month. The viral RNA titers of infants born to GBV-C/HGV-infected mothers appeared as elevated as those of their mothers. All the GBV-C/HGV-infected infants remained positive for viral RNA during the entire study period. No clinical events possibly linked to a primary GBV-C/HGV infection were reported in infants. Serum ALT level and blood count remained within normal values throughout the follow-up of all GBV-C/HGV infected infants. CONCLUSION: The frequency of mother-to-infant GBV-C/HGV transmission is elevated and could explain the high prevalence of GBV-C/HGV markers (viral RNA and E2 antibody) in adults at low risk for blood-borne or sexually transmitted viruses, such as blood donors. PMID- 10827268 TI - John Elliott and the evolution of American blood banking, 1934 to 1954. PMID- 10827269 TI - HLA antibodies after transfusions of FFP. PMID- 10827270 TI - The risk of transfusion transmission of Chagas disease in Mexico City. PMID- 10827271 TI - Transfusion-related acute lung injury and granulocytes from stored packed RBCs. PMID- 10827272 TI - Lack of macroscopic aggregates in pools of prestorage WBC-reduced random-donor platelet concentrates prepared from whole blood. PMID- 10827273 TI - Molecular genetic analysis of the Japanese amorph rh(null) phenotype. PMID- 10827274 TI - The concentration of AS-1 RBCs after inverted gravity sedimentation for neonatal transfusions. PMID- 10827275 TI - Genomic DNA fingerprinting of Oenococcus oeni strains by pulsed-field gel electrophoresis and randomly amplified polymorphic DNA-PCR. AB - Genetic diversity of 60 Oenococcus oeni strains from different wines was evaluated by numerical analysis of (i) pulsed-field gel electrophoresis (PFGE) patterns with endonuclease ApaI and (ii) randomly amplified polymorphic DNA (RAPD)-PCR fingerprints with four oligonucleotide primers. Sixty-two percent of the strains could be distinguished by PFGE, whereas most strains were identified by distinct RAPD-PCR profiles and associated according to the geographical origin. Because of its rapidity and reliability, RAPD-PCR appeared to be a suitable method for typing and monitoring O. oeni strains in winemaking. PMID- 10827276 TI - Diversity of cyanobacterial hydrogenases, a molecular approach. AB - In an effort to elucidate the diversity of cyanobacterial hydrogenases, we used a molecular approach. Filamentous strains from a broad range of sources were screened for the presence of hup (uptake hydrogenase), xisC (rearrangement within hupL), and hox (bidirectional hydrogenase) genes. As expected, an uptake hydrogenase seems to be present in all N(2)-fixing cyanobacteria. On the other hand, no evidence was found for the presence of a conventional bidirectional enzyme in several strains. Similarly, the presence of xisC is not a characteristic shared by all the heterocyst-forming cyanobacteria. Although tempting, it is not possible to establish a correlation between the presence/absence of the bidirectional hydrogenase and the occurrence of xisC. The natural molecular variation of hydrogenases in cyanobacteria is certainly a field to explore, both to understand the physiological functions of the respective enzymes and to identify a genetic background to be used when constructing a strain for photobiological H(2) production in a bioreactor. PMID- 10827277 TI - Isolation of a gene from Burkholderia cepacia IS-16 encoding a protein that facilitates phosphatase activity. AB - A genomic library from Burkholderia cepacia IS-16 was constructed in Escherichia coli by partial Sau3AI digestion of the chromosomal DNA, with the plasmid vector Bluescript SK. This library was screened for clones able to grow as green stained colonies on selective medium developed for detecting phosphatase-positive colonies. Three green-stained clones (pFS1, pFS2, and pFS3) carried recombinant plasmids harboring DNA inserts of 5.0, 8.0, and 0.9 kb, respectively. DNA hybridization experiments demonstrated the presence of overlapping DNA fragments in the three clones and that these three clones were all derived from Burkholderia cepacia IS-16 genomic DNA. DNA sequence analysis, together with polyacrylamide gels of proteins encoded by E. coli containing pFS3, suggested that the isolated 0. 9-kb DNA fragment encodes the functional portion of a phosphate transport protein. PMID- 10827278 TI - Transformation of internal extracellular bacteria isolated from Rhagoletis completa cresson gut with enhanced green fluorescent protein. AB - We discovered Zeocintrade mark is an effective antibiotic against Enterobacter agglomerans and Klebsiella pneumoniae strains isolated from the walnut husk fly (Rhagoletis completa Cresson: Family Tephritidae) and that bleomycin resistance can be used as a selective marker in transforming plasmids. We transformed Ent. agglomerans and K. pneumoniae strains originally isolated from their close association with R. completa gut to produce enhanced green fluorescent protein, a variant of green fluorescent protein in the first demonstration of genetic transformation of internal extracellular bacteria isolated from a tephritid pest. We report methods for plasmid-mediated transformation of these bacteria, the expression of fluorescent marker protein from the transforming plasmids, and the stability of the transforming plasmid in the bacteria. We also discuss applications of this technology in the study of pest biology and control implementation. PMID- 10827279 TI - Genomic rearrangements in neisseria meningitidis strains of the ET-5 complex. AB - A physical map of the chromosome of Neisseria meningitidis strain 44/76, which belongs to the epidemic clone ET-5, was constructed. DNA fragments obtained after SfiI and NheI digestion were resolved by pulsed field gel electrophoresis (PFGE). The overall arrangement of 26 genetic markers localized on the 2.3-Mb chromosome was conserved in comparison with that in meningococcal strains B1940 and Z2491. Simplified physical maps of 29 additional strains belonging to the ET-5 complex isolated from various parts of the world were compared with that of strain 44/76. Ten distinct patterns of hybridization were identified. While two of the seven probes hybridized to fragments of the same size in all strains, the remaining probes hybridized to different fragments, in some cases to fragments not adjacent on the chromosome of 44/76. These results indicated the occurrence of genetic rearrangements in the genome of the ET-5 meningococcal clone in the course of its epidemic spread. 30 years now. A large number of social science questions, however, remain to be answered. In this article, we review the rapidly growing but geographically lopsided literature on this topic. Using computer database searches supplemented by reference reviews and professional correspondence with those active in the field, we gathered literature on the social science and service delivery aspects of emergency contraception published in English up through December 1998, as well as a few unpublished papers from the same time and slightly later, representing regions where published material is practically nonexistent. Methodologically acceptable papers are summarized in our tables and text, and form the basis for suggested improvements in existing emergency contraceptive services. The review also offers ideas for designing new emergency contraception services where they do not yet exist. We conclude by proposing an agenda for further social science research in this area. PMID- 10827332 TI - Prolonged effectiveness of Norplant(R) capsule implants: a 7-year study. AB - Soft tubing Norplant(R) contraceptive implants were studied in 1210 women for 7 years to measure the duration of effectiveness and the magnitude of the pregnancy rates over that time. Mean age at enrollment was 27.4 years. Of the enrollees, 42% were US residents. One-sixth (16.1%) weighed >/=70 kg at the time of implant placement. At the end of 5 years, the cumulative pregnancy rate was 1.1/100; at the end of 7 years, it was 1.9/100. No pregnancies occurred to any of the 400 women who enrolled in the study at age >/=30 years and who weighed <100 kg. Among women aged 18-33 years, the 7-year Norplant pregnancy rates are comparable to the median pregnancy rates of tubal sterilization methods for women of the same age and duration of use. For women aged >/=34 years, without regard to weight at admission, the 7-year effectiveness of soft tubing Norplant equals or surpasses that of tubal sterilization. For continuing implant users, annual pregnancy rates <1.0/100 in years 6 and 7, together with low cumulative pregnancy rates, testify that Norplant capsule implants remain highly effective for 7 years. PMID- 10827333 TI - Comparison of ovarian follicular activity during treatment with a monthly injectable contraceptive and a low-dose oral contraceptive. AB - Ovarian follicular development occurs during treatment with combined and progestin-only oral contraceptive (OC) pills and progestin-containing subdermal implants, and can be associated with the development of persistent functional cysts that may require surgical removal. Lunelle is a once-a-month injectable contraceptive containing estradiol cypionate 5 mg and medroxyprogesterone acetate 25 mg. A randomized, comparative study was undertaken to compare the effect on ovarian follicular activity associated with use of Lunelle and a low-dose OC. A total of 30 ovulatory subjects were randomly assigned to receive two cycles of treatment with either an OC containing ethinyl estradiol 20 microg and 0.1 mg levonorgestrel or Lunelle. During the second cycle of treatment, pelvic sonography was performed every 4 days, at which time the maximum follicle diameter was measured. Study end points were the presence of follicles >/=10, 20, and 30 mm. In all, 13 of 15 subjects in the OC group and 14 of 15 in the Lunelle group completed the study. Follicles measuring >/=10 mm were present in 11 of 13 (84.6%) in the OC users and in four of 14 (28.6%) in the Lunelle users (p <0.05). In the OC group, six of 13 subjects (46.1%) developed follicles >/=20 mm, and one of 13 (7.7%) developed follicles >/=30 mm. No subjects in the Lunelle group developed a follicle >/=20 mm in diameter. This study indicates that Lunelle is associated with a significantly lower incidence of ovarian follicular development compared to that of an OC containing 20 microg ethinyl estradiol and 0.1 mg levonorgestrel. PMID- 10827334 TI - Ovarian function during use of Nestorone(R) subdermal implants. AB - Nestorone(R) progestin (NES) is a potent 19-nor-progesterone derivative which is biologically inactive when administered orally; however, it is an excellent option for implant contraception. The objective of this study was to evaluate ovarian function during use of either one 4-cm or two 3-cm NES implants for 24 months. A total of 60 volunteers were enrolled in each dose group. Vaginal ultrasound (VUS) and blood sampling for determinations of estradiol (E(2)), progesterone (P) and NES serum levels were carried out twice a week for 6 consecutive weeks, beginning in months 1, 6, 12, 18, and 24 of implant use. Serum levels of NES declined with time, with a more pronounced decrease during the first 18 months of implant use; thereafter, NES levels remained stable until the end of the study at 24 months. Luteal activity was very infrequent during the first year of use (<3%) but increased during the second year, occurring in 27% and 35% of the sampling periods in the 1-implant group, and 2% and 16% of the sampling periods in the 2-implant group, at months 18 and 24 of use, respectively. No luteal activity was observed with NES levels above 80 pmol/L. Serum P levels in periods of luteal activity were significantly lower than those of controls. Persistent anovulatory follicles were the most common VUS finding and this was associated with E(2) levels that remained within the normal range (101-1500 pmol/L) in the majority of the sampling periods studied. Considering that a single implant offers advantage for insertion and removal, a new single NES implant is being developed with a slightly higher release rate, to reduce effectively the incidence of ovulation and provide a greater margin of safety beyond 2 years. PMID- 10827335 TI - Comparative evaluation of three Tactylon(TM) condoms and a latex condom during vaginal intercourse: breakage and slippage. AB - This study compared breakage and slippage rates of three male condom styles made of Tactylon(TM), a synthetic elastomer, to those of a marketed latex condom during vaginal intercourse. Safety and acceptability outcomes were also assessed. This two-center, prospective, crossover study enrolled 443 couples. Each couple was randomly assigned to use three condoms of each type in one of 24 use sequences. Couples completed questionnaires after using each condom, all of one condom type, and all four condom types. The percentage and standard error (SE, in parentheses) of latex condoms with clinical breakage was 0.86% (0.295). Percentages for Tactylon condoms were not equivalent to the latex study condom, ranging from 3.50% to 4.17%. The percentage and SE of latex condoms with complete slippage was 1.11% (0.328). Percentages for Tactylon condoms were equivalent to those for latex, ranging from 0.70% to 1.31%. The Low-Modulus Tactylon condom was the most preferred. Fewer medical events were reported with the Tactylon condoms than with the latex condom. It was concluded that Tactylon condoms were equivalent to the latex condom in terms of slippage but not breakage. However, safety and acceptability seemed to be better for Tactylon condoms. This may improve consistency of use and may attract new users. PMID- 10827336 TI - Safety and acceptability of a baggy latex condom. AB - A total of 104 couples participated in a randomized crossover trial to compare a new baggy condom with a straight-shaft condom produced by the same manufacturer. Participants completed a coital log after using each condom. All couples used five condoms of each type. Among 102 couples who did not report major deviations from the protocol, the breakage rate was eight of 510 (1.6%) for the baggy condom, and six of 510 (1.2%) for the standard condom (rate difference, RD = 0. 4%, 95% confidence interval of the RD, CI = -1.0%; +1.8%). Slippage was reported in 50 baggy condom logs and in 58 standard condom logs; the slippage rate was 50 of 510 (9.8%) for the baggy condom, and 58 of 510 (11.4%) for the standard condom (RD = -1.6%, 95% CI = -5.4%; +2.2%). Slippage was most often partial (<1 inch) and may not indicate condom failure. Severe slippage rates were 11 of 510 (2.2%) for the baggy condom, and 18 of 510 (3.5%) for the standard condom (RD = -1.4%, 95% CI = -3.4%; +0.7%). The findings support the conclusion that the two condoms are equivalent with respect to breakage and slippage. The participants appeared to prefer the baggy condom, suggesting that the new product may be more acceptable to the public than the traditional straight-shaft condoms, and may be easier to use consistently over long time periods. PMID- 10827337 TI - Predictors of analgesia use during supervised medical abortion. The Mifepristone Clinical Trials Group. AB - The object of this analysis was to identify predictors of narcotic analgesic use during medical abortion. A total of 2121 women with pregnancies of phosphatidylethanolamine and PC > LPE/EPL > phosphatidylinositol > PS. Further, CLA increased the proportions of n-3 fatty acids in the individual glycerophospholipids. High CLA diets containing 3% and 5% of a CLA mixture were associated with increased activity of catalase in the peroxisome-enriched cell fraction of liver and exhibited marked reductions of cholesterol in the low and high density lipoproteins relative to rats receiving no CLA. PMID- 10827341 TI - Effects of phospholipids on sphingomyelin hydrolysis induced by intestinal alkaline sphingomyelinase: an in vitro study. AB - Digestion of dietary sphingomyelin (SM) is catalyzed by intestinal alkaline sphingomyelinase (SMase) and may have important implications in colonic tumorigenesis. Previous studies demonstrated that the digestion and absorption of dietary SM was slow and incomplete and that the colon was exposed to SM and its hydrolytic products including ceramide. In the present work, we studied the influences of glycerophospholipids and hydrolytic products of phosphatidylcholine (PC; i.e., lyso-PC, fatty acid, diacylglycerol, and phosphorylcholine) on SM hydrolysis induced by purified rat intestinal alkaline SMase in the presence of 10 mM taurocholate. It was found that various phospholipids including PC, phosphatidylserine (PS), phosphatidylinositol (PI), phosphatidylethanolamine (PE), and phosphatidic acid (PA) inhibit alkaline SMase activity in a dose dependent manner, with the degree of inhibition being in the order PA > PS > PI > PC > PE. Similar inhibition was also seen in a buffer of pH 7.4, which is close to the physiologic pH in the middle of the small intestine. When the effects of hydrolytic products of PC were studied, lyso-PC, oleic acid, and 1,2-dioleoyl glycerol also inhibited alkaline SMase activity, whereas phosphorylcholine enhanced SMase activity. However, in the absence of bile salt, acid phospholipids including PA, PS, and PI mildly stimulated alkaline SMase activity whereas PC and PE had no effect. It is concluded that in the presence of bile salts, glycerophospholipids and their hydrolytic products inhibit intestinal alkaline SMase activity. This may contribute to the slow rate of SM digestion in the upper small intestine. PMID- 10827342 TI - Myristic acid, unlike palmitic acid, is rapidly metabolized in cultured rat hepatocytes. AB - This study was designed to examine and compare the metabolism of myristic and palmitic acids in cultured rat hepatocytes. [1-(14)C]-Labeled fatty acids were solubilized with albumin at 0.1 mmol/L in culture medium. Incubation with 24-hr cultured hepatocytes was carried out for 12 hr. Myristic acid was more rapidly (P < 0.05) taken up by the cells than was palmitic acid (86.9 +/- 0.9% and 68.3 +/- 5.7%, respectively, of the initial radioactivity was cleared from the medium after 4 hr incubation). Incorporation into cellular lipids, however, was similar after the same time (33.4 +/- 2.8% and 34.9 +/- 9.3%, respectively, of initial radioactivity). In the early phase of the incubation (30 min), myristic acid was more rapidly incorporated into cellular triglycerides than was palmitic acid (7.4 +/- 0.9% and 3.6 +/- 1.9%, respectively, of initial radioactivity). However, after 12 hr incubation, the radioactivity of cellular triglycerides, cellular phospholipids, and secreted triglycerides was significantly higher with palmitic acid as precursor. Myristic acid oxidation was significantly higher than that of palmitic acid (14.9 +/- 2.2% and 2.3 +/- 0.6%, respectively, of the initial radioactivity was incorporated into the beta-oxidation products after 4 hr). Myristic acid was also more strongly elongated to radiolabeled palmitic acid (12.2 +/- 0.8% of initial radioactivity after 12 hr) than palmitic acid was to stearic acid (5.1 +/- 1.3% of initial radioactivity after 12 hr). The combination of elongation and beta-oxidation results in the rapid disappearance of C14:0 in hepatocytes whereas C16:0 is esterified to form glycerolipids. This study provides evidence that myristic acid is more rapidly metabolized in cultured hepatocytes than is palmitic acid. PMID- 10827343 TI - JNK1 is inactivated during thiamine deficiency-induced apoptosis in human neuroblastoma cells. AB - Thiamine deficiency results in selective neuronal damage. A number of mechanisms have been proposed to account for brain damage associated with thiamine deficiency and to account for the focal nature of the loss of neurons. One proposed mechanism is programmed cell death. We found efficient induction of apoptosis in human neuroblastoma cells when the cells were deprived of thiamine. Although extensive mitochondrial damage was seen, the release of cytochrome c was not the triggering mechanism for thiamine deficiency-induced apoptosis. Instead, the activity of the cJun amino terminal kinase Jnk1 was lost, and this loss correlated temporally with induction of apoptosis. The loss was specific for Jnk1; Jnk2/3 activity remained unchanged. Loss of Jnk1 activity was not found in lymphoblasts, a cell type that did not undergo apoptosis when deprived of thiamine. These findings suggest that thiamine deficiency results in a cellular stress that brings about the loss of Jnk1 activity and the loss of its function of protecting cells from programmed cell death. We postulate that focal sensitivity to thiamine deficiency results, in part, from specific neuronal cell types being susceptible to the inactivation of Jnk1 in response to depletion of cellular thiamine. PMID- 10827344 TI - Green tea polyphenols (flavan 3-ols) prevent oxidative modification of low density lipoproteins: an ex vivo study in humans. AB - Oxidation of low density lipoprotein (LDL) plays crucial roles in atherogenesis. We previously reported that green tea polyphenols (flavan 3-ols), especially epigallocatechingallate (EGCg) and epicatechingallate, exerted potent inhibitory effects on LDL oxidation in vitro. To examine whether intake of green tea polyphenols renders LDL resistant to ex vivo oxidation in humans, 22 male volunteers aged between 22 and 32 years were recruited and assigned the same dietary regimen for 2 weeks. After a 1-week baseline period, they were equally divided into two groups: control and tea. The tea group ingested 300 mg of green tea polyphenol extract twice daily for 1 week. Plasma EGCg concentration at the end of the experiment was 56 nmol/L on average (56% in free form) in the tea group; no EGCg was detected before the experiment. Plasma concentrations of lipids, ascorbate, alpha-tocopherol, and lipid peroxides did not change before and after the experiment in either group, but beta-carotene was higher in the tea group (P< 0.01 by paired Student'st-test). LDL (0.1 mg/mL) was incubated with 5 microM Cu(2+) and the oxidation was measured by absorbance at 234 nm. The lag time was significantly prolonged by 13.7 min in the tea group (P < 0.05 by paired Student'st-test, before versus after), whereas such a change was not observed in the control group. These results suggest that daily consumption of seven to eight cups (approximately 100 mL each cup) of green tea may increase resistance of LDL to in vivo oxidation, leading to reduction in the risk of cardiovascular diseases. PMID- 10827345 TI - Indicators of erythrocyte formation and degradation in rats with either vitamin A or iron deficiency. AB - Vitamin A deficiency produces anemia and altered iron status. In this study with rats we tested two hypotheses regarding vitamin A deficiency: (1) that it impairs erythropoiesis, leading to an increased red cell turnover, and (2) that it inhibits the glycosylation of transferrin. Erythropoietic activity was assessed indirectly by determining the myeloid:erythroid ratio in bone marrow smears, the number of erythroid colonies in the red pulp of spleen, the blood reticulocyte index, and zinc protoporphyrin and plasma transferrin receptor concentrations. Transferrin glycosylation was assessed by measuring the sialic acid content of transferrin. The effects of vitamin A deficiency were compared with those of iron deficiency. Iron deficiency produced anemia and low iron levels in organs. Vitamin A deficiency produced low levels of plasma and hepatic retinol, and it induced decreased plasma total iron-binding capacity and raised iron levels in tibia and spleen. Short- but not long-term iron deficiency reduced the number of erythroid colonies in spleen; vitamin A deficiency had no influence. Neither iron nor vitamin A deficiency influenced the myeloid:erythroid ratio in bone marrow smears and the blood reticulocyte production. Plasma transferrin receptor and erythrocyte zinc protoporphyrin concentrations were not affected by vitamin A deficiency but increased with iron deficiency. Vitamin A deficiency did not stimulate erythrocyte breakdown, as indicated by unaltered plasma lactate dehydrogenase activity and reduced plasma total bilirubin levels. Both vitamin A and iron deficiencies raised the proportion of multiple sialylated transferrins in plasma. Thus, we have not found evidence that vitamin A deficiency affects erythropoiesis and erythrocyte turnover. The iron accumulation in spleen and bone marrow may be related to reduced iron transport due to inhibition of transferrin synthesis rather than inhibition of transferrin sialylation. PMID- 10827346 TI - Dietary coconut oil affects more lipoprotein lipase activity than the mitochondria oxidative capacities in muscles of preruminant calves. AB - The presence of coconut oil in a milk replacer stimulates the growth rate of calves, suggesting a better oxidation of fatty acid in muscles. Because dietary fatty acid composition influences carnitine palmitoyltransferase I (CPT I) activity in rat muscles, this study was designed to examine the effects of a milk replacer containing either tallow (TA) or coconut oil (CO) on fatty acid utilization and oxidation and on the characteristics of intermyofibrillar (IM) and subsarcolemmal (SS) mitochondria in the heart and skeletal muscles of preruminant calves. Feeding CO did not affect palmitate oxidation rate by whole homogenates, but induced higher palmitate oxidation by IM mitochondria (+37%, P < 0.05). CPT I activity did not significantly differ between the two groups of calves. Heart and longissimus thoracis muscle of calves fed CO had higher lipoprotein lipase activity (+27% and 58%, respectively; P < 0.05) but showed no differences in fatty acid binding protein content or activity of oxidative enzymes. Whatever the muscle and the diet, IM mitochondria had higher respiration rates and enzyme activities than those of SS mitochondria (P < 0.05). Furthermore, CPT I activity of the heart was 28-fold less sensitive to malonyl coenzyme A inhibition in IM mitochondria than in SS mitochondria. In conclusion, dietary CO marginally affected the activity of the two mitochondrial populations and the oxidative activity of muscles in the preruminant calf. In addition, this study showed that differences between IM and SS mitochondria in the heart and muscles were higher in calves than in other species studied so far. PMID- 10827347 TI - [Telomerase and cancer: a therapeutic breakthrough or a cul-de-sac?]. PMID- 10827348 TI - [An uncommon tumor of the urinary bladder: the small cell carcinoma]. AB - Primary small cell carcinoma of the urinary bladder is an uncommon tumor, compared to the frequency of urothelial tumors. Fifty percent of cases are combined with a non endocrine carcinomatous component. We report six new cases of this tumor, three of pure, and three associated with an urothelial carcinoma. Diagnosis is easy established by the immunohistochemical study which show the neuro-endocrine differentiation of these aggressive tumors. Pathologist needs to look for a neuro-endocrine part in all bladder cancer, as its presence modify the treatment. Chemotherapy is used in these cancers, due to their high metastatic power. Places of radical surgery and radiotherapy need to be specified. PMID- 10827349 TI - [Standards, Options and Recommendations (SOR): nutritional support in oncohematology]. PMID- 10827350 TI - [Standards, Options and Recommendations (SOR) for the use of appetite stimulants in oncology. Work group. Federation of the French Cancer Centres (FNCLCC)]. AB - CONTEXT: The "Standards, Options and Recommendations" (SOR) project, started in 1993, is a collaboration between the Federation of the French Cancer Centres (FNCLCC), the 20 French Cancer Centres and specialists from French Public Universities, General Hospitals and Private Clinics. The main objective is the development of clinical practice guidelines to improve the quality of health care and outcome for cancer patients. The methodology is based on literature review and critical appraisal by a multidisciplinary group of experts, with feedback from specialists in cancer care delivery. OBJECTIVES: To develop clinical practice guidelines according to the definitions of the Standards, Options and Recommendations project for the use of appetite stimulants in oncology. METHODS: Data were identified by searching Medline and the personal reference lists of members of the expert groups. Once the guidelines were defined, the document was submitted for review to 55 independent reviewers, and to the medical committees of the 20 French Cancer Centres. RESULTS: The main recommendations for the use of appetite stimulants in oncology are: 1) Corticosteroids and the synthetic progestogens (megestrol acetate and medroxyprogesterone acetate) are appetite stimulants. 2) They can be useful in managing anorexia and weight loss in cancer patients, especially in the palliative setting, despite the potential side effects of these agents. 3) The most effective way of using these drugs is not known. Inclusion in clinical trials is recommended. 4) Cyproheptadine, metoclopramide, nandrolone and pentoxif line should not be used outside prospective clinical trials. 5) Hydrazine sulfate should not be used. PMID- 10827351 TI - [Percutaneous endoscopic gastrostomy: cost/benefit analysis in patients with carcinoma of the upper aero-digestive tract]. AB - The aim of this retrospective study was to evaluate the cost and benefit of percutaneous fluoroscopic gastrostomy feeding (PFG) in 70 cancer patients with advanced stage disease of the upper-aero digestive tract; we retrospectively analyzed the consequences in terms of nutritional status (evaluated by weight and body mass index), the possibility to lead a treatment by high dose chemo radiotherapy to the end of the therapeutic schedule, the feasibility, complications and cost ratios. Three weeks after the procedure, no major complication was observed, the initial nutritional threshold was conserved. PFG is a safe and effective technique; the additional cost is low (2%) compared with the total cost of hospitalization. PMID- 10827352 TI - [++Scintimammography: a new imaging technique for diagnosis and follow-up of breast cancer]. AB - 99m Tc MiBi has emerged as a new imaging technique for primary breast carcinomas. The aim of this study was to analyze the diagnostic performance and the additional information provided by scintimammography (SMN). Two hundred and forty consecutive women referred to surgery for abnormalities breast or suspicious breast cancer were scanned before surgery. Sensitivity and specificity were respectively 92% and 53%. PPV: 77.5% and PVN: 79.3%. All false negative scans occurred in cancers 1 cm. SMN detected multiple foci of uptake in the same breast in 13%, that were all confirmed to be multifocal disease and histology. Controlateral focal uptake was also detected; at this time 5/27 are confirmed to be bilateral breast neoplasms; 22 patients are in follow-up. SMN is reliable in the diagnosis of breast cancer and also with difficult cases of mammography. Moreover, SMN provides additional qualitative information in 19.6% of breast carcinomas, such as chest wall infiltration, multifocal or bilateral breast cancer. PMID- 10827353 TI - [Case report forum: the example of French sarcoma group multidisciplinary discussion tools. Sarcomes de la FNCLCC]. AB - Within the Rubis 4th framework of European project is led a pilot experiment of tools and services for health professionals in prospect for the Aquitanian healthcare network. The sarcoma group of the FNCLCC (47 people) uses on its web site a multidisciplinary dialogue with a specific discussion forum. This service allows the anonymous publication of a imaging clinical case and to start a take care discussion. 87 cases were published in 13 months involving 261 answers from February 1999 to February 2000. A case is published every 4 days on average and the deadlines for replies regularly drop (15 days in February 1999 down 1.1 day in February 2000). The cases are published either for a diagnosis or treatment request (30%) or for the physical preparation of meeting or for the continuous medical training (70%). There are many advantages in comparison with the other possibilities of discussion: availability, autonomy of publication, cost, number of experts participating. These NTIC services will be developing within the regional healthcare oncology networks and are already tested by other regional groups (Lymphoma) considering the simplicity of use, management and training of the functionality. PMID- 10827354 TI - [Evaluation of the first social consultation in oncology: a series of 200 cases. Groupe de recherche de l'action medico-sociale en Aquitaine (Gramsa)]. AB - The aim of the present study is to evaluate the clinical social work in oncology. We present a series of 137 men and 63 women treated for cancer (mean age: 57 y; 20-90) who were addressed (65%) or came spontaneously (35%) for a first social consultation; 45 consultations concerned the family at the day of death of the patient. The collected data (expressed requests, evaluated real difficulties, orientations of the social work, necessity of network) are correlated with the medical data and the time between diagnosis and social consultation. The social intervention is often brief (95%) and focused (66%). A medical and social network is used in 70% of cases. The administrative and psychosocial problems predominate with an important discordance between the requests and the evaluated difficulties. The metastatic status increases psychosocial, legal difficulties and problems of the organisation of home-care. The delay of the social consultation has great influence upon the legal difficulties. The family's difficulties, after the patient's death are administrative (93.3%), psychosocial (84.4%) and legal (68.9%). Clinical social work is part of the global management of patients and their families; its quality has direct implication on their daily life during and after treatment. PMID- 10827355 TI - A support group for children with one parent with cancer: report on 4 year experience of a talking group. AB - Children of a parent diagnosed with cancer, experience severe emotional distress, which may have important repercussions later, in adult life. However, responses of children to a parent with a serious illness vary, depending on the functioning of the family unit, and more especially, the capacity of the family to communicate openly and effectively about the parent's illness. A support group has now been established at IGR for children of parents undergoing treatment for cancer, in order to help foster intra-familial communication about coping with illness. The group is open to children and other family members and consists of a non-structured discussion of approximately 2 hours. The support group is animated by a psychologist and a doctor. In 4 years, 175 children, aged from 6 months to 23 years, from 98 families have participated in the programme. The emotional and behavioural conduct of the children and their family were analysed. Children were encouraged to express their fears concerning their parent's illness and age appropriate medical information was given in response to their questions. As a result of this opportunity to express and share feelings with others, children's anxiety was shown to be considerably lessened and a renewal of discussion within the family followed in the majority of cases. Nevertheless, the dynamics of these groups can be delicately balanced. It is therefore paramount that the animation of such groups be confined to experienced professionals. PMID- 10827356 TI - [Do health workers practice what they preach? A study of knowledge, opinions and practices of health care professionals, with respect to their own health, in Madagascar]. AB - In this study, a questionnaire was used to study the individual behavior of a sample of fifty health workers, doctors and nurses from Tamatave (Madagascar) with respect to a number of current health issues. We used the results to assess the impact and efficacy of previous training and to obtain information about the extent to which health staff actually follow the recommendations and standards that they are given and expected to teach, as far as their own health is concerned. It was clear that the standard management procedures and instructions concerning basic health-preserving behavior were far from being universally accepted and followed by health workers themselves. This was the case even for common diseases and health issues for which specific training had been given, such as malaria, diarrhea, AIDS and STDs, tobacco use, HBP management and the use of antibiotics. More attention should be paid to involving health workers in their training programs so that they are really convinced by the recommendations given. So long as most health workers are not themselves committed to the recommendations and have inadequate behavior regarding their own health, they are unlikely to be effective at promoting good health practice. PMID- 10827357 TI - [Nutrition problems experienced by adults infected with the human immunodeficiency virus in Abidjan (Ivory Coast)]. AB - Denutrition is frequent among HIV-infected (HIV+) adults in sub-Saharan Africa. One of the risk factors for denutrition is a reduction in dietary intake. Eating disorders may be partly responsible for such decreases in food intake. We prospectively analyzed the frequency, associated factors and progression of anorexia, dysphagia and food aversion in a cohort of 330 HIV-infected adults included in a trial of early chemoprophylaxis with cotrimoxazole in Abidjan, Ivory Coast. Patients were followed-up by means of scheduled monthly visits. Eating disorders were assessed using a standardized questionnaire after 6, 12 and 18 months of follow-up. After six months of follow-up, 28% of the patients reported anorexia, 9% dysphagia and 28% food aversion. Multivariate analysis showed that anorexia was significantly more frequent in women than in men (odds ratio (OR) = 2.0 [95% confidence interval: 1.2-3.5]) and in patients with a CD4+ lymphocyte count < 200/mm3 (OR = 1.8 [1.0-3.5]). The risk of dysphagia was also higher for women than for men (OR = 1.8 [1.0-3.5]). The risk of dysphagia was also higher for women than for men (OR = 3.3 [1.3-8.4]). Patients with < 200 CD4+ lymphocytes/mm3 were more likely than those with CD4+ lymphocyte counts of over 200 to suffer food aversion (OR = 1.8 [1.1-3.0]). We analyzed the progression of dietary problems during follow-up and found that anorexia and dysphygia tended to disappear from one evaluation to the next whereas the number of patients reporting food aversion tended to increase. For patients reporting anorexia at the 6-month evaluation, significantly more women than men reported the persistence of anorexia at the 12-month evaluation (16% versus 5%; p = 0.03). Among patients with dysphagia at six months, those with a CD4+ lymphocyte count below 200/mm3 were much more likely than those with a CD4+ count above 200 to report persistent dysphagia at the 12-month evaluation (7% versus 0%; p = 0. 02, Fischer's exact test). For patients with no dietary problems after six months, those taking cotrimoxazole were significantly more likely than those of the placebo group to report food aversion at the 12-month evaluation (21% versus 8%; p = 0.01). We found that dietary problems were associated more with the stage of immunodeficiency that with socioeconomic factors, with the exception of sex, which was associated with several outcomes. These data stress the importance of detecting these frequent dietary problems as part of the overall clinical management of HIV+ adults in Africa, and of providing affected individuals with early nutritional counseling. PMID- 10827358 TI - [Hygiene, working conditions and professional risks in the Moorish "hammams++" in Marrakech]. AB - Moorish "Hammam" baths are used by almost all of the Moroccan population. We evaluated the occupational hazards associated with these baths by studying the working conditions, hygiene and safety at ten Hammam baths in Marrakech. We carried out a descriptive, cross-sectional epidemiological study of 60 workers (30 men and 30 women). We analyzed working conditions, hygiene and safety and carried out a medical survey by means of a questionnaire, clinical tests, chest X rays and biological screening. The working environment in Hammam baths is particularly hot and humid. We found working conditions, hygiene and safety to be poor. Elementary regulations concerning environmental health were not respected and no means of protection was provided for the staff. Mycological studies showed that fungi pathogenic for humans were present in 100% of floor samples. Bacteriological analysis of the well water supplying the baths and of the residual water collected from various bath chambers showed that pathogenic bacteria resistant to multiple antibiotics were present. The medical survey identified various diseases affecting staff members, mostly infectious in nature. The most common problems observed were: problems muscular or skeletal in nature or linked to posture (28.3%), dermatological (100%), neurological (71.6%), ocular (30%), respiratory (90%), ear/nose/throat (51.6%), digestive (11.6%), oral (63.3%), gynecological (83.3%) and urinary (31.6%). Often, individual workers suffered several morbidities associated with working conditions simultaneously In light of these major occupational hazards, it is clearly important to implement special medical supervision of workers in Hammam baths, to improve hygiene and working conditions and to ensure the occupational health education and training of those concerned. As a first step, an occupational health service should be set up in the offices of the municipal health department. PMID- 10827359 TI - [Appearance on X-ray and computed tomography of femoral head necrosis in patients with sickle cell disease]. AB - The aim of this study was to describe X-ray and computed tomography signs and to estimate the severity of femoral head necrosis in patients with sickle cell disease. We studied two groups of patients with sickle cell disease (group A: 14 patients with no hip pain and group B: 34 patients with hip pain) over 17 months. Each patient underwent hip X-ray (lying face-down) and CT scan. None of the patients in group A presented hip lesions whereas we detected 42 necrotic femoral head lesions by standard X ray and 51 by CT scan. The lesions were frequently bilateral (82.14%) and were more frequent in young patients (from 11 to 30 years old). Their progression was faster in patients with the SS genotype. No significant difference was observed according to type of hemoglobin. All patients with sickle cell disease presenting with hip pain should undergo CT scan to check for femoral head necrosis. PMID- 10827360 TI - [++Helicobacter pylori infection in North Lebanon]. AB - The aim of this study was to determine the prevalence of Helicobacter pylori (HP) infection in North Lebanon and to compare it with published figures. The study period was from September 1 1996 to March 31 1998. We tested 349 patients for HP, using biopsy tissue obtained by esophagoduodenoscopy and the modified urease technique. One hundred and seventy-nine of these patients were men and 170 were women. The mean age of the subjects was 39.3 years (range: 20 to 78 years). The prevalence of HP was 25.9% in individuals with normal endoscopy results (n = 104), 60.8% in patients with duodenal ulcer and 72% in patients with congestive antritis. The prevalence of HP in patients with erosive antritis was similar to that in individuals with normal endoscopy results. The prevalence of HP increased with age in the individuals with pathological endoscopic results but no such relationship was observed in individuals with normal endoscopy results. Our results show that the prevalence of HP in our patients is similar to that of industrialized countries. As age affected the prevalence of HP only in subjects with pathological endoscopy results, it is probably the extent of lesions caused by HP, rather than the infection itself, that increases with age. PMID- 10827362 TI - [Endometrial carcinomas in Gabon. A study of 34 cases in 11 years: 1988-1998]. AB - The aim of this retrospective study was to determine the importance of endometrium cancer in Gabon over an eleven-year period (1988-1998). We studied 34 cases from the register of cancers at the Faculty of Medicine in Libreville. The incidence of endometrium cancers was 5.3%. The patients came from areas with medical care facilities. They were all mature women with a mean age of 58.8 years (range: 34 to 80 years). Most of these women were menopausal (88. 24%). Uterine bleeding was the main symptom (100% of cases). Histologically, endometrium adenocarcinomas (79.41%) were the most frequently observed. The setting up of medical facilities in rural areas and efforts to provide the population with information concerning these cancers may affect the incidence and prognosis of endometrium cancer in Gabon. PMID- 10827361 TI - [Severely burned patients: epidemiology and treatment (a study of 104 Gabonese cases)]. AB - This retrospective study was carried out over five years (August 1993 to August 1998) and included 104 patients admitted to the intensive care unit for heat induced or electrical burns affecting more than 10% of their total body surface area. Most of the patients were children or young adults and the mean age of the group was 24 years. Seventy-eight of the patients were the victims of domestic fires. The other 26 cases involved work-related burns and car accidents. Most of the burns observed were caused by fire or scalding, but there were also two cases of electrical burns. Lesions affected predominantly the head (45.1%), upper limbs (31.5%) and perineum (5.8%). Hemodynamic rescuscitation and intensive respiratory care were administered initially, along with topical surgical treatment. Triple antibiotic treatment was also given immediately in cases of shock or burns to the body's natural orifices. If triple antibiotic treatment was not administered immediately then, within six hours of the burn, treatment was given to prevent infection with Staphylococcus aureus and soil-borne microorganisms and anti tetanus vaccination was adminstered systematically. The treatment was then modified to prevent infection with Gram-negative bacilli, common second-stage microoganisms. The bacteria most frequently isolated, particularly from the skin and urine, were Pseudomonas (52%), Escherichia coli (37.5%) and Staphylococcus aureus (10.5%). Enteral and parenteral nutrition was begun as soon as possible. The presence of the patient's family during care and rehabilitation was of great psychological benefit to the patients. The mean duration of hospital stay was 12 days. In those cases in which the patient died, early death (within one week) was due to respiratory distress and hydroelectrolytic problems whereas deaths after the first week were due to septic shock. The overall death rate was 54.8%. Prevention should be taught, with particular emphasis on those at high risk. PMID- 10827363 TI - [Drug-resistant malaria: problems with its definition and technical approaches]. AB - In antimalarial chemotherapy, drug resistance is defined as "the ability of a parasite strain to survive and/or multiply despite the administration and absorption of a drug in doses equal to or higher than those usually recommended but within the limits of tolerance of the subject". This official World Health Organization definition, based on clinical and parasitological observations, was established in 1973, when genetics, pharmacology and in vitro culture techniques were still in the early stages of development. Several techniques are currently used to detect drug-resistant Plasmodium falciparum. Several in vivo tests, the traditional gold standard for the detection of drug resistance, have been developed. Classical tests include the 28-day extended test and the 7-day test, interpreted using the S-RI-RII-RIII classification system (S for susceptible and R for resistant, with three degrees of resistance, I to III, depending on parasitological response). These tests cannot be applied in practice, in field situations, and the results do not take into account the clinical condition of the patient, largely because they were designed for use with asymptomatic carriers. These limitations led to the development in 1994 (modified in 1996) of the more practical and simplified 14-day test of therapeutic efficacy. This test classifies the patient's clinical and parasitological response as "adequate clinical response", "late treatment failure" or "early treatment failure". This in vivo test of therapeutic efficacy can be applied in the field with a minimum of health facilities, personnel and other resources. However, true cases of drug resistance may not always be detected by in vivo tests due to pharmacokinetic variations, reinfection, multiple infections, noncompliance or interference with the acquired immune response. The most commonly used reliable in vitro assay, the isotopic microtest, determines the drug concentration at which 50% of parasite growth is inhibited (50% inhibitory concentration IC50). The in vitro assay not only yields quantitative results, it also determines the phenotype of the parasite independently of the immune and physiopathological conditions of the host. However, this in vitro assay requires highly skilled personnel and laboratory equipment. In addition, parasites isolated from patients who have taken medication on their own initiative a few days before consultation usually do not grow in vitro and the interpretation of assay results for patients with multiple infections may be equivocal. One of the major problems with in vitro tests is the determination of the threshold IC50 values that distinguish susceptible from resistant parasites. There are currently no fully validated cut off points for assessing in vitro resistance. Despite these shortcomings, in vitro tests are of value, particularly if performed in parallel with the in vivo test. Molecular biology has made a major contribution to our understanding of the mechanisms of drug resistance. Discrete point mutations in the genes encoding dihydrofolate reductase and dihydropteroate synthase are strongly associated with resistance in vitro to pyrimethamine and sulfadoxine, respectively. Preliminary results have also suggested that these mutations are responsible for the failure of sulfadoxine-pyrimethamine combination treatment. No causal relationship between discrete polymorphisms in the candidate genes and in vitro chloroquine resistance has yet been established. High-performance liquid chromatography is being increasingly used to determine the plasma concentrations of antimalarial drugs in patients with prophylactic or therapeutic failure, to check that the failure of the treatment is not due to inadequate levels of the drug in the patient. Taking into account all these aspects of resistance to antimalarial drugs we think that the WHO definition of drug resistance is now inadequate. (ABSTRACT TRUNCATED) PMID- 10827364 TI - [Foreign international reference databases: MEDLINE and PubMed. Presentation and instructions for use]. AB - The aim of this paper is to provide a French guide to reference retrieval using MEDLINE and the new version of PubMed, a system for retrieving references from the World Wide Web. We explain, screen-by-screen, how this reference retrieval tool works, to make its use easier for French-speaking end-users. PMID- 10827365 TI - [Rapid trachoma assessment method (TRA): comparison with an exhaustive prevalence survey in a region of endemic trachoma in Mali]. AB - In 1997, the World Health Organization (WHO) endorsed a new strategy aimed at controlling blinding trachoma by 2020. The WHO developed a rapid assessment method for trachoma (TRA) for identifying communities affected by blinding trachoma and for ranking villages with endemic trachoma in order of priority for intervention. We applied this method to communities in Mali in which trachoma was prevalent, and compared the results with those of a classical prevalence survey. Seven villages from the Dogon region were first investigated using the rapid method. Members of the local community were asked to identify individuals with suspected trichiasis, using a diagram to help them with recognition. The individuals thus identified were then examined by an ophthalmic nurse for confirmation of the diagnosis. Fifty children aged less than ten years from poorer households were then selected. These children underwent ophthalmic examination, using a magnification of 2.5, and trachoma status was established. The next day, the villages were visited by an ophthalmologist who examined all of the inhabitants. The rapid assessment method identified 18 of the 52 patients requiring surgery. This method had a sensitivity of 34.6% and a specificity of 98%. The prevalence of active trachoma among children was 29.0% in the exhaustive survey and 29.9% for the rapid assessment method. Overall sensitivity was 54.6% and overall specificity 79.6%, as the two methods did not identify the same individuals. The Kappa coefficient, estimating agreement between observers, was 0.34. The ranking of the villages in order of priority differed slightly for the two methods. With the rapid assessment method, five of the seven villages were classified as having a prevalence of over 20% whereas, with the exhaustive survey, six of the seven villages fell into this category. TRA was not designed to replace epidemiological survey and the prevalence of trachoma should not be inferred from this method. However, it is a useful tool for determining the order of priority for intervention of communities at risk. PMID- 10827366 TI - [Treatment of rheumatoid polyarthritis with methotrexate in Dakar: efficacy, tolerance and cost]. AB - Methotrexate (MTX), which has been used for years in cancer treatment, is now being proposed as a first-line treatment for rheumatoid arthritis (RA), despite its potential side effects. The aim of this study was to investigate the short term efficacy, safety and relative cost of low-dose MTX for the treatment of RA. We carried out an open, nonrandomized trial in which patients received a 7 mg injection of MTX once per week, with clinical and biological follow up. A single physician performed the weekly assessments, which involved evaluation of the duration of morning stiffness, the number of night awakenings, the number of painful and swollen joints and Ritchie's index. Blood cell count and erythrocyte sedimentation rate were determined monthly. Twelve RA patients were enrolled in the trial, over a mean treatment period of 356 +/- 175 days. A significant improvement was observed in all variables except the number of swollen joints. Ritchie's index decreased from a mean of 31.8 +/- 11.85 to 6.5 +/- 8.98 (p<1.6 x 10- 4). Minor adverse reactions were observed but none indicated treatment withdrawal: 6 cases of nausea, 2 of a moderate increase in transaminase activity, 1 of bronchitis, in which the responsibility of MTX was not definitely established and 3 cases in which hemoglobin levels decreased. The monthly cost of the treatment, including the drug itself and laboratory tests, is lower than that of gold salt injection. Three issues of key importance in our region were investigated in this study: 1) the possible desire to become pregnant of female patients undergoing MTX treatment. In addition, some of the young and unmarried patients did not understand or appreciate the contraceptive effects of the treatment; 2) poor compliance with the treatment due to limited financial resources. Many patients did not regularly attend for their follow-up appointments and many stopped taking the medication. One third of the patients were lost to follow-up during this study; 3) the prevalence of chronic hepatitis, which may limit the use of MTX in our region. Serological tests should be performed before the treatment is started and a liver biopsy is recommended for patients with chronic hepatitis B or C. PMID- 10827367 TI - Transesophageal echocardiography guided enoxaparin antithrombotic strategy for cardioversion of atrial fibrillation: the ACUTE II pilot study. AB - BACKGROUND: Patients with atrial fibrillation >2 days' duration for whom immediate cardioversion is desired or required are commonly hospitalized for 4 or more days of antithrombotic therapy with intravenous unfractionated heparin and commencement of oral warfarin. For these early cardioversion patients, self administered low-molecular-weight heparin (enoxaparin sodium) as "bridge" therapy to warfarin may obviate the need for hospitalization and activated partial thromboplastin time monitoring and thus potentially lower costs and enhance utility. OBJECTIVE: To compare feasibility and safety of a transesophageal echocardiography (TEE)-guided enoxaparin strategy with those of a TEE-guided unfractionated heparin strategy in patients with atrial fibrillation of >2 days' duration undergoing early electrical or chemical cardioversion. DESIGN AND SETTING: This is a randomized, multicenter clinical trial at 11 hospitals in the United States. PATIENTS AND INTERVENTION: Two hundred patients with atrial fibrillation >2 days' duration requiring early chemical or electric cardioversion will be enrolled. TEE-guided intravenous unfractionated heparin bridge therapy will be compared with TEE-guided subcutaneous enoxaparin bridge therapy. OUTCOME MEASURES: Feasibility outcomes are time to hospital discharge, patient quality of life/utility, treatment costs, and sinus rhythm. Safety outcomes are ischemic stroke, transient ischemic attack, systemic embolization, major and minor bleeding, clinical hemodynamic instability, and cardiac and cardioversion-related death for a 5-week period from enrollment. CLINICAL IMPLICATIONS: The results of this pilot study will have important clinical and economic implications for the antithrombotic management of patients with atrial fibrillation undergoing TEE guided cardioversion. PMID- 10827368 TI - High-dose bolus lidocaine for chemical cardioversion of atrial fibrillation: a prospective, randomized, double-blind crossover trial. AB - BACKGROUND: Most drugs used for chemical cardioversion of atrial fibrillation have significant proarrhythmia risk and require close monitoring after administration. Lidocaine has few of the proarrhythmic concerns of most antiarrhythmic drugs and, at high bolus doses, prolongs the atrial refractory period well enough to be effective in converting atrial fibrillation to sinus rhythm. This finding has been previously demonstrated in a dog model. We sought to confirm the animal findings in human beings with lidocaine doses of 1.5 to 2.5 mg/kg. METHODS: Twenty patients with atrial fibrillation scheduled for elective cardioversion were enrolled in this study. In a randomized, double-blind, crossover study design, each patient received intravenous bolus lidocaine or saline. Patients were observed for 10 minutes after the initial bolus to assess efficacy. The second test drug was then delivered if the first was unsuccessful at cardioversion. RESULTS: All 20 patients received both lidocaine and saline placebo therapy in a crossover manner. None of the 20 patients converted to sinus rhythm with either therapy. The 95% confidence interval for effectiveness of lidocaine in this population was 0% to 14%. CONCLUSION: In this population of patients referred for elective cardioversion of atrial fibrillation, high-dose bolus lidocaine was ineffective in converting patients to sinus rhythm. Although this study was not sufficiently powered to rule out a low efficacy of lidocaine (<15%) or a higher efficacy in certain subgroups of atrial fibrillation, routine use of lidocaine for this indication is not warranted. PMID- 10827369 TI - Design of the blockade of the glycoprotein IIb/IIIa receptor to avoid vascular occlusion (BRAVO) trial. AB - BACKGROUND: Platelets play a key role in the pathogenesis of atherosclerosis, thrombosis, and acute coronary and cerebrovascular syndromes. Inhibition of platelet function by acetylsalicylic acid (aspirin) has been shown to reduce the incidence atherothrombotic events in patients with coronary, cerebrovascular, or peripheral vascular disease. Thienopyridine agents, however, including ticlopidine and clopidogrel, inhibit the adenosine diphosphate receptor and have modestly superior effects compared with aspirin on reduction of death, myocardial infarction, and stroke among a broad group of patients with vascular disease. More effective antithrombotic agents are still required to treat patients at high risk for recurrent vascular events. METHODS: Lotrafiban, a selective, nonpeptide antagonist of the human platelet fibrinogen receptor (glycoprotein [GP] IIb/IIIa [alphaIIb/beta3 integrin]), blocks the binding of fibrinogen to the GP IIb/IIIa receptor, which is the final common pathway of platelet aggregation. Lotrafiban at doses of up to 50 mg twice daily was well-tolerated in a 12-week, double blind, placebo-controlled, dose-ranging study in patients with recent myocardial infarction, unstable angina, transient ischemic attack, or stroke when added to aspirin therapy. On the basis of these results, a dosing regimen was selected for the phase III Blockage of the Glycoprotein IIb/IIIa Receptor to Avoid Vascular Occlusion (BRAVO) trial based on pharmacodynamics and drug tolerability. In the pivotal BRAVO study, lotrafiban therapy is being evaluated in patients who have had a recent myocardial infarction, unstable angina, transient ischemic attack, or ischemic stroke, or who present at any time after a diagnosis of peripheral vascular disease combined with either cardiovascular or cerebrovascular disease. RESULTS: The efficacy evaluation will be based on a composite end point of clinical events (death by any cause, myocardial infarction, stroke, recurrent ischemia requiring hospitalization, or urgent ischemia-driven revascularization). The target enrollment is 9200 patients worldwide. Approximately 700 centers will participate and will be distributed within 30 countries across North America, Europe, Australia, and Asia. PMID- 10827370 TI - Influence of proteinuria on long-term outcome among patients with diabetes: the evidence continues to accumulate. PMID- 10827371 TI - Endovascular stent treatment of aortic coarctation. PMID- 10827372 TI - Proteinuria is a key determinant of death in patients with diabetes after isolated coronary artery bypass grafting. AB - BACKGROUND: Proteinuria is a marker for underlying diabetic nephropathy and may be a surrogate marker for advanced atherosclerosis. It is unknown if proteinuria is a determinant of death in patients with diabetes after coronary artery bypass grafting. We hypothesized that diabetic patients with evidence of proteinuria would have increased mortality and clinical event rates after isolated coronary artery bypass grafting compared with nonproteinuric diabetic patients. METHODS AND RESULTS: We performed an observational of study of 905 diabetic patients with urinalysis and available follow-up data (nonproteinuria, n = 651; proteinuria, n = 254) after isolated coronary artery bypass grafting at the Cleveland Clinic Foundation between January 1989 and December 1992. The proteinuria group was further prospectively stratified into low-concentration (n = 225) and high concentration (n = 29) groups. The end points of this study were all-cause mortality and the composite end point of death, nonfatal myocardial infarction, and need for repeat revascularization. The mean follow-up time was 66 months. The 5-year mortality rate for the nonproteinuria and proteinuria groups was 20.2% and 29.1% (P <.001), respectively. The 5-year rate of death, nonfatal myocardial infarction, and need for repeat revascularization for the nonproteinuria and proteinuria groups was 25.2% and 36.2% (P <.001), respectively. Significant multivariate predictors of 5-year mortality included age, not using a left internal mammary artery graft to the left anterior descending coronary artery, proteinuria, lower body weight, and increased creatinine level. CONCLUSIONS: Among diabetic patients, proteinuria appears to be an important predictor of death after isolated coronary artery bypass grafting. PMID- 10827373 TI - Clinical information determines the impact of transesophageal echocardiography on the diagnosis of infective endocarditis by the duke criteria. AB - BACKGROUND: Although transesophageal echocardiography (TEE) is more sensitive than transthoracic echocardiography (TTE) in detecting echocardiographic evidence of infective endocarditis (IE), the impact of TEE on the clinical diagnosis of IE has not been clearly delineated. METHODS AND RESULTS: We studied 112 patients with 114 suspected episodes of IE over a 6-year period who underwent both TTE and TEE during their diagnostic evaluation. Using the results of these studies along with clinical and microbiologic data, we attempted to determine the incremental value of TEE to the Duke Endocarditis Diagnostic Criteria. Patients were initially classified into a diagnostic category of the Duke criteria with TTE data, and then the diagnostic classification was reconsidered with TEE data. A diagnostic category reassignment occurred in 25 of 114 episodes of IE evaluated when TEE results were incorporated into the evaluation with the Duke criteria (22 patients were reclassified from possible IE to definite IE whereas 3 patients were reclassified from rejected to possible IE). Diagnostic reclassification occurred in 9 (11%) of the 80 episodes of suspected IE with native cardiac valves and 13 (34%) of 34 episodes with prosthetic cardiac valves. Most patients reclassified from possible IE to definite IE with TEE data (19 of 22) had an intermediate clinical likelihood of IE, whereas 92% of patients had negative TTE results. Pathologic examination of valvular tissue in 22 of the 114 episodes of suspected IE revealed that the positive predictive value of the Duke criteria with TEE data for diagnosis of IE was 85% in patients with native valves and 89% in patients with prosthetic valves. CONCLUSIONS: When clinical evidence of IE is present, TEE improves the sensitivity of the Duke criteria to diagnose definite IE. TEE data appears to be especially useful for the diagnostic evaluation of patients with suspected IE who have prosthetic valves. PMID- 10827374 TI - Subgroups, treatment effects, and baseline risks: some lessons from major cardiovascular trials. AB - BACKGROUND: The objective of this study was to determine how subgroup analyses are performed in large randomized trials of cardiovascular pharmacotherapy. METHODS AND RESULTS: We reviewed 67 randomized, double-blind, controlled trials involving pharmacotherapy in at least 1000 patients with unstable angina, myocardial infarction, left ventricular dysfunction, or heart failure with clinical outcomes as primary end points, published between 1980 and 1997. Nine had no subgroup analyses but 43 reported on 5 or more subgroups and 31 reported subgroups without formal statistical tests for treatment-subgroup interactions. In most trials, a rationale for subgroup selection was missing. All but 6 focused on single-factor subgroups. CONCLUSIONS: Trial subgroups should ideally be defined a priori on 2 bases: single-factor subgroups with a strong rationale for biological response modification and multifactorial prognostic subgroups defined from baseline risks. However, single-factor subgroup analyses are often reported without a supporting rationale or formal statistical tests for interactions. We suggest that clinicians should interpret published subgroup-specific variations in treatment effects skeptically unless there is a prespecified rationale and a significant treatment-subgroup interaction. PMID- 10827375 TI - Predictors of recurrent ischemic events and death in unstable coronary artery disease after treatment with combination antithrombotic therapy. AB - BACKGROUND: Patients with non-Q-wave acute coronary syndromes (ACS) have substantial rates of recurrent ischemic events, but prognostic studies have been small or preceded the routine use of aggressive combination antithrombotic therapy. We sought to identify predictors of these events after antithrombotic treatment of non-Q-wave ACS. METHODS: We assessed 30-day rates of a composite triple end point (death, infarction, or refractory angina) and double end point (death or infarction) among 3171 patients with non-ST-segment elevation ACS randomly assigned to enoxaparin or heparin, plus aspirin, for 2 to 8 days. We created multivariable regression models to predict these end points from baseline factors. RESULTS: Overall, 682 patients (21%) reached the triple end point and 220 (6.8%) reached the double end point. Independent predictors of the triple end point were admission with myocardial necrosis, ST-segment depression, prior angina severity, symptom duration, and allocation to enoxaparin treatment in patients with ST-segment depression (significant interaction). Independent predictors of the double end point were admission with myocardial necrosis, ST segment depression, enrollment region, age >75 years, prior angina severity, and rales. By deciles, the average predicted risk for the double end point ranged from 2% to 20%: a patient aged <75 years with no risk factors had a 3.5% risk, whereas a patient aged >75 years with 2 additional high-risk features (myonecrosis and ST depression) had a risk of death or reinfarction of 26%. CONCLUSIONS: Patients with non-ST-segment elevation ACS exhibit a broad range of risk of adverse recurrent ischemic events. The predictive power of the model for the triple end point, using baseline variables, was modest. However, a subgroup at very low risk of the double end point (average 2%) can be identified with baseline variables. PMID- 10827376 TI - Clinical importance of risk factors and exercise testing for prediction of significant coronary artery stenosis in women recovering from unstable coronary artery disease: the Stockholm Female Coronary Risk Study. AB - BACKGROUND: The objectives of this study were to investigate the relation between coronary risk factors, exercise testing parameters, and the presence of angiographically significant coronary artery disease (CAD) (> or =50% luminal stenosis) in female patients previously hospitalized for an acute CAD event. METHODS AND RESULTS: All women younger than age 66 years in the greater Stockholm area in Sweden who were hospitalized for acute coronary syndromes during a 3-year period were recruited. Besides collection of clinical parameters, coronary angiography and a symptom-limited exercise test were performed in 228 patients 3 to 6 months after the index hospitalization. The mean age was 56 +/- 7 years. Angiographically nonsignificant CAD (stenosis <50%) was verified in 37% of the patients; significant CAD was found in 63%. The clinical parameters that showed the strongest relation with the presence of significant CAD after adjusting for age were history of myocardial infarction (odds ratio [OR] 4.91, 95% confidence interval [CI] 2.35 to 7.49), history of diabetes mellitus (OR 3.83, 95% CI 1.63 to 14.31), serum high-density lipoprotein cholesterol <1.4 mmol/L (OR 2.11, 95% CI 1. 20 to 3.72), and waist-to-hip ratio >0.85 (OR 1.78, 95% CI 1.02 to 3. 10). A low exercise capacity and associated low change of rate-pressure product from rest to peak exercise were the only exercise testing parameters that were significantly related to angiographically verified significant CAD (<90% of the predicted maximal work capacity adjusted for age and weight, OR 1.91, 95% CI 1. 04 to 3.50). CONCLUSIONS: In female patients recovering from unstable CAD, exercise capacity was the only exercise testing parameter of value in the prediction of significant CAD. The consideration of certain clinical characteristics and coronary risk factors offer better or complementary information when deciding on further coronary assessment. PMID- 10827377 TI - Acute myocardial infarction in young adults: prognostic role of angiotensin converting enzyme, angiotensin II type I receptor, apolipoprotein E, endothelial constitutive nitric oxide synthase, and glycoprotein IIIa genetic polymorphisms at medium-term follow-up. AB - BACKGROUND: A number of reports have investigated the association between various gene polymorphisms and the phenotypic expression of myocardial infarction. No investigations have evaluated the prognostic role of genetic factors in young people with premature coronary disease. The aim of this study was to investigate the influence of genetic factors compared with that of conventional risk factors on follow-up events in a population of Italian young adults with myocardial infarction. METHODS AND RESULTS: The study population consisted of 106 young patients (mean age 40 +/- 4 years, range 23 to 45 years) with diagnosis of acute myocardial infarction. Clinical and genetic data from the group of patients with events during follow-up were compared with those from patients without events. The following genetic polymorphisms were tested: angiotensin I converting enzyme, angiotensin II type I receptor, apolipoprotein E (ApoE), endothelial constitutive nitric oxide synthase, and platelet glycoprotein IIIa. Coronary angiography was performed in 94 patients. Coronary angiography showed coronary artery disease in 93% of patients. During follow-up (46 +/- 12 months, range 25 to 72) the overall combined end points (cardiac death, myocardial infarction, and revascularization procedures) accounted for 21 events. Family history of coronary artery disease, smoking, stenosis of the left anterior descending artery at coronary angiography, and ApoE polymorphism (presence of epsilon4 allele) were significantly more prevalent (univariate analysis) in the group of patients with events. Logistic multivariate analysis showed that ApoE polymorphism (P =. 004, odds ratio [OR] 6.8, 95% confidence interval [CI] 2 to 22), family history (P =.005, OR 8.3, 95% CI 2 to 35), smoking after acute myocardial infarction (P =.008, OR 10.9, 95% CI 2 to 62), and left anterior descending coronary artery disease (P =.02. OR 6.6, 95% CI 1.3 to 33) were independent predictors of adverse events. CONCLUSIONS: Myocardial infarction at a young age is commonly characterized by evidence of multiple cardiovascular risk factors and by a favorable prognosis in short- and medium-term follow-up. Evidence of significant disease at coronary angiography suggests the presence of a premature atherosclerotic process. ApoE polymorphism (presence of epsilon4 allele) appears to be a strong independent predictor of adverse events, suggesting a remarkable influence in the accelerated coronary disease. PMID- 10827378 TI - Factors associated with delay in reperfusion therapy in elderly patients with acute myocardial infarction: analysis of the cooperative cardiovascular project. AB - BACKGROUND: Many elderly patients with an acute myocardial infarction (AMI) do not receive thrombolysis within 30 minutes of hospital arrival as recommended by the American College of Cardiology/American Heart Association Guidelines. We sought to identify factors associated with delay in administration of thrombolysis after arrival to the hospital in these patients and to determine whether this delay is associated with increased mortality rates. METHODS AND RESULTS: By using the Cooperative Cardiovascular Project database, we identified patients who received thrombolysis for an AMI. The patients were stratified into groups by time to thrombolysis after hospital arrival. Among a cohort of 17,379 patients, 22.2% received thrombolysis in the first 30 minutes after hospital arrival. Patients treated after the first 30 minutes were more likely to be older, be female, be diabetic, have a history of hypertension or heart failure, and have less marked ST elevation. They were also more likely to be admitted to smaller hospitals with a lower volume of AMIs and to hospitals without a cardiac catheterization laboratory. The 30-day mortality rate was significantly lower for patients treated within the first 30 minutes. After adjustments were made for clinical and hospital characteristics, delays in therapy beyond 30 and 90 minutes were associated with an increase in 1-year mortality rates of 9% and 27%, respectively, compared with delays for patients treated within 30 minutes. CONCLUSIONS: After hospital arrival, time to treatment with thrombolytic therapy is longer than recommended in a significant proportion of patients. Clinical characteristics and institutional factors are associated with the delay in treatment. The more rapid treatment of appropriate elderly patients with an AMI probably will reduce mortality rates. PMID- 10827379 TI - Serial quantitative coronary angiography and coronary events. AB - BACKGROUND: Although assessment of progression of atherosclerosis by quantitative coronary angiography (QCA) is used as a surrogate for coronary events, no validation study has compared the several QCA measures used. METHODS AND RESULTS: The Cholesterol Lowering Atherosclerosis Study was a clinical trial testing the efficacy of colestipol-niacin on the progression of coronary atherosclerosis. Baseline/2-year coronary angiograms were obtained on 156 men with prior coronary artery bypass graft surgery. Changes in percent diameter stenosis and minimum lumen diameter (both measured in coronary lesions and segments) and coronary segment measures of average diameter, percent involvement, and vessel edge roughness were measured by QCA. Coronary events ascertained over 12 years of follow-up included myocardial infarction (MI), coronary death, and coronary artery revascularizations. Proportional hazards models evaluated the relation between QCA change measures and coronary events. Changes in percent diameter stenosis and minimum lumen diameter of coronary artery lesions were significantly related to the risk of MI/coronary death. All QCA measures were significantly related to the risk of any coronary event. Relative risks for each QCA measure were of similar magnitude when estimated separately within each treatment group. Change in minimum lumen diameter of lesions was the only measure independently associated with the risk of coronary events. CONCLUSIONS: All QCA measures of progression of coronary artery disease were related to all coronary events (including revascularizations). Only QCA measures of lesion progression were related to MI/coronary death. QCA measures of lesion change may be better surrogate end points for "hard" coronary events than measures of change in coronary segments. PMID- 10827380 TI - Current status of prevention, diagnosis, and management of coronary artery disease in patients with kidney failure. AB - BACKGROUND: Patients with kidney failure have a heavy burden of coronary artery disease. The results of preventive, diagnostic, and therapeutic measures developed in nonuremic populations cannot automatically be extrapolated to this unique group of patients. METHODS AND RESULTS: Articles were reviewed if they contained English language text or an abstract identified by MEDLINE search from 1980 to 1999, supplemented by manual review of bibliographies of published articles and abstract issues of national cardiology meetings, studies on diagnostic techniques, risk modification measures, pharmacologic agents, and coronary revascularization procedures in patients with uremia. Descriptive and quantitative data as appropriate were extracted. Lipid-lowering agents may be safely administered to uremic patients. Direct evidence of lipid lowering in this population is not available and is not likely to be forthcoming. Erythropoietin therapy is effective in reversing the cardiovascular perturbations of uremic anemia, but an approach of normalizing the hematocrit cannot be recommended. Glycoprotein IIb/IIIa inhibitors used in acute coronary syndromes require downward dose adjustment or are contraindicated. Thrombolytic agents are underutilized in the management of myocardial infarction. Noninvasive testing is less accurate than in nonuremic populations. Coronary revascularization offers relative clinical advantages over medical therapy similar to non-kidney failure populations, even though the results in uremic patients is significantly less favorable than for nonuremic patients. Stenting is the preferred revascularization approach, and conventional balloon percutaneous transluminal coronary angioplasty the least favorable. CONCLUSIONS: Many but not all of the benefits of therapies developed in nonuremic patients extend to patients with kidney failure. Physicians should be familiar with the advantages and limitations of each of these modalities in this population. PMID- 10827381 TI - Sex-specific triggers for right ventricular outflow tract tachycardia. AB - BACKGROUND: Right ventricular outflow tract tachycardia (RVOT-VT) is a common arrhythmia in young patients without heart disease. The arrhythmia is characterized by repetitive bursts and premature ventricular contractions with a left bundle branch block, inferior-axis QRS morphology, and symptoms of palpitations. Although more frequent in women, sex-specific triggers for symptomatic RVOT-VT have not been identified. METHODS AND RESULTS: We interviewed 34 women and 13 men referred for ablation of RVOT-VT to determine if predictable but sex-specific exacerbations in symptomatic RVOT-VT exist. After a general query asking if there was predictability to what triggered palpitations, we then specifically queried all patients about symptomatic RVOT-VT initiation with exercise, stress, caffeine, fatigue, and, in women only, periods of recognized hormonal flux. The times identified as states of hormonal flux included premenstrual, gestational, perimenopausal, and coincident with the administration of birth control pills. In response to the completed interview, the most common recorded trigger for RVOT-VT in women was recognized states of hormonal flux with 20 (59%) of 34 women responding positively and 14 (41%) of the 34 indicating that states of hormonal flux were the only recognizable triggers. Men were more likely than women to report that their RVOT-VT was predictably triggered by exercise, stress, or caffeine: 12 (92%) of 13 men versus 14 (41%) of 34 women (P <.01). CONCLUSIONS: Triggers for RVOT-VT initiation are sex specific. Women have RVOT-VT initiation with recognized states of hormonal flux. Men more commonly have RVOT VT initiated by exercise or stress. These data have important implications related to patient education and counseling in the setting of RVOT-VT and may influence the timing of drug treatment and electrophysiologic evaluation in selected patients. PMID- 10827382 TI - Changes over time in the incidence and case-fatality rates of primary ventricular fibrillation complicating acute myocardial infarction: perspectives from the Worcester Heart Attack Study. AB - BACKGROUND: Limited population-based data are available that describe temporal and recent trends in the incidence and case-fatality rates in patients with primary ventricular fibrillation (VF) complicating acute myocardial infarction (AMI). The purpose of this study was to describe changes over a 22-year period (1975 through 1997) in the incidence and hospital case-fatality rates of primary VF complicating AMI from a multihospital, community-wide perspective. METHODS AND RESULTS: This was an observational study of metropolitan Worcester residents hospitalized with a validated uncomplicated AMI (n = 5020) in all hospitals in the Worcester, Massachusetts, metropolitan area (1990 census population = 437,000) during 11 1-year periods between 1975 and 1997. The overall incidence rate of primary VF complicating AMI was 4.7%. The crude as well as multivariable adjusted odds of the development of VF did not change significantly over the 22 year period under study. The overall in-hospital case-fatality rate of patients with primary VF was 44%, which was significantly greater in comparison with AMI patients in whom VF did not develop (5%). Hospital mortality rates associated with primary VF declined over time. Improved survival was observed in patients who had primary VF in the 1990s after adjusting for potential prognostic confounders. CONCLUSIONS: The results of this community-wide study failed to indicate changes over time in the incidence rates of primary VF in patients hospitalized with AMI between 1975 and 1997. On the other hand, hospital death rates in patients with primary VF have shown encouraging declines during more recent periods. These mortality trends are likely to be the results of improvements in the treatment and more careful surveillance of patients with AMI. PMID- 10827383 TI - Continuous monitoring of an endocardial index of myocardial contractility during head-up tilt test. AB - BACKGROUND: Previous studies suggest that vigorous myocardial contractions stimulate ventricular mechanoreceptors and lead to vasovagal syncope. We studied an endocardial index of myocardial contractility during the head-up tilt test in vasovagal patients and control patients, and we evaluated the effect of negative inotropic drugs on myocardial contractility and tilt test outcome. METHODS AND RESULTS: We investigated 19 patients with recurrent vasovagal syncope and positive tilt test (group 1) and 11 patients with no syncope and negative tilt test (group 2). Myocardial contractility was continuously measured during a tilt test (60 degrees ) through a microaccelerometer incorporated in the tip of a right ventricular electrode to sense left ventricular contractility. Patients in groups 1 and 2 were evaluated during an unmedicated tilt test, and patients in group 1 were reevaluated during a tilt test with infusion of esmolol (n = 10) or disopyramide (n = 9). During the unmedicated test, patients in group 1 exhibited a significant increase in myocardial contractility immediately on postural change (P <.05), unlike patients in group 2. Patients in group 1 also had a further increase in myocardial contractility before the end of tilt (P <.01). With drug administration, the changes in supine myocardial contractility were nonsignificant and were not related with the outcome of the tilt test (P <.05). CONCLUSIONS: An increase in myocardial contractility is detected by the sensor during the tilt test. The changes induced by the drugs on supine myocardial contractility are minor and not related with the outcome of the head-up tilt test. PMID- 10827384 TI - Prediction of death after percutaneous coronary interventional procedures. AB - BACKGROUND: The prediction and comparison of procedural death after percutaneous coronary interventional procedures is inherently difficult because of variations in case mix and practice patterns. The impact of modern, expanded patient selection criteria, and newer technologic approaches is unknown. Our objective was to determine whether a risk equation based on patient-related variables and derived from an independent data set can accurately predict procedural death after percutaneous coronary intervention in the current era. METHODS AND RESULTS: An analysis was made of the Mayo Clinic Coronary Interventional Database January 1, 1995, to October 31, 1997. Expected mortality rate was calculated with the use of the New York State multivariate risk score. In 3387 patients, 3830 procedures (55.1% stents) were performed, with an expected mortality rate of 2.32% and observed mortality rate of 2.38% (P = not significant). The risk score derived from the New York multivariate model was highly predictive of death (chi-square = 213.8; P <.0001). The presence of a high-risk lesion characteristic such as calcium, thrombus, or type C lesion was modestly associated with death. CONCLUSIONS: The New York State multivariate model accurately predicted procedural death in our database. PMID- 10827385 TI - Adjunctive intracoronary dipyridamole in the interventional treatment of small coronary arteries: a prospectively randomized trial. AB - BACKGROUND: Patients undergoing intracoronary stent placement or balloon angioplasty for the treatment of small coronary arteries are at an increased risk of an adverse outcome from a higher incidence of abrupt closure and restenosis. Intracoronary thrombus formation plays a key role in the pathogenesis of abrupt vessel closure and of restenosis. Dipyridamole prevents platelet aggregation by a mechanism that differs from aspirin. The purpose of this study was to investigate the effect of intracoronary dipyridamole on acute complications and restenosis after percutaneous transluminal coronary angioplasty. METHODS: In a prospectively randomized study including 491 dilatations of coronary arteries with a diameter <2.75 mm, additional intracoronary application of dipyridamole was compared with conventional pretreatment consisting of heparin and aspirin. Study end points were defined as incidence of abrupt vessel closure, myocardial infarction, angiographic restenosis, and target vessel revascularization rate. RESULTS: Intracoronary dipyridamole was associated with a significant reduction of abrupt vessel closure (2.8% vs 8.6%; P =.005) and a nonsignificant reduction of myocardial infarction (1.6% vs 4.5%; P =.07) after percutaneous transluminal coronary angioplasty. Net gain 6 months after angioplasty was significantly higher in the dipyridamole group (0.60 +/- 0.35 mm vs 0.42 +/- 0.34 mm; P <.001). However, dipyridamole failed to reduce the incidence of angiographic restenosis (41.6% vs 49.1%; P =.11) and target vessel revascularization rate (20.6% vs 269%; P =.12). CONCLUSIONS: Intracoronary dipyridamole reduces the incidence of adverse cardiovascular events in the first 48 hours after balloon angioplasty of small coronary arteries. Reduction of restenosis rates failed to reach statistical significance. However, a significant increase in net gain was observed. Thus intracoronary application of dipyridamole should be considered in the treatment of small coronary arteries when intracoronary stenting is not appropriate. PMID- 10827386 TI - Review of immediate angioplasty after fibrinolytic therapy for acute myocardial infarction: insights from the RESCUE I, RESCUE II, and other contemporary clinical experiences. AB - BACKGROUND: Prompt restoration of Thrombolysis In Myocardial Infarction (TIMI) grade 3 flow improves survival in patients with acute ST-segment elevation myocardial infarction (MI). Fibrinolytic therapy fails to restore TIMI 3 flow within 90 minutes in 40% to 50% of patients. Because the results of percutaneous coronary intervention (PCI) for MI seem to be improving, a reevaluation of the role of PCI after fibrinolytic therapy for MI appears to be warranted. METHODS AND RESULTS: Data from all 9 randomized controlled trials (including new data from 4 trials) of rescue percutaneous transluminal coronary angioplasty (PTCA) versus conservative therapy after fibrinolytic therapy (1456 patients), 4 contemporary registries of PCI in this setting (977 patients), and other germane studies are reviewed. PTCA after failed fibrinolysis (TIMI 0 to 1 flow) appears to reduce early severe heart failure (3. 8% vs 11.7%, P =.04) and improve survival over 1 year in patients with moderate to large MI (92% vs 87%, P =.001) and possibly reduces early repeat MI (4.3% vs 11.3%, P =.08). Assessment of the possible benefit of PTCA for TIMI 2 flow is hampered by the small number of patients randomly assigned. Repeat MI may be decreased and left ventricular functional recovery enhanced. PTCA early after successful fibrinolysis is nearly always technically successful and may reduce repeat MI and hospital length of stay. However, it must be recalled that randomized trials from the 1980s suggested increased mortality rates with PTCA after restoration of TIMI 2 to 3 flow with fibrinolysis. Data from contemporary randomized studies of stents and glycoprotein IIb/IIIa inhibitors suggest that PCI as performed today may yield better results than those reviewed. CONCLUSIONS: These data suggest a probable benefit of rescue PTCA in several distinct scenarios and that the pivotal mid 1980s studies suggesting no benefit or harm for PTCA after fibrinolytic therapy may no longer be relevant. The role of mechanical intervention in the treatment of patients treated in these settings should be reassessed. PMID- 10827387 TI - Early results and medium-term follow-up of stent implantation for mild residual or recurrent aortic coarctation. AB - BACKGROUND: Optimal timing and mode of treatment for patients with aortic coarctation remains controversial, particularly when the degree of obstruction is mild. Surgery, balloon dilatation, and stent implantation have all proven effective in the treatment of moderate or severe obstruction. In this report, we describe the use of stents to treat coarctation in a heterogeneous population, including patients with relatively mild obstruction. METHODS: Retrospectively, we studied the results of stent implantation in 33 patients, children and young adults, who underwent catheterization for treatment of coarctation. Patients with isolated coarctation, as well as those with associated cardiac defects, were included. The median systolic pressure gradient of our patients was 25 mm Hg. RESULTS: Patients had an acute decrease in systolic blood pressure gradient (25 mm Hg to 5 mm Hg, P <.001) and an increase in lumen diameter (8 mm to 13 mm, P <.001). When 16 patients were recatheterized during the follow-up period, gradients remained decreased (30 mm Hg to 14 mm Hg, P <.001) compared with prestent values. Ventricular end-diastolic pressure, which was increased in 82% of patients at the time of initial catheterization, decreased from 17 mm Hg to 14 mm Hg (P =.002). Although the procedure was generally safe, serious complications did occur. CONCLUSIONS: Stent implantation represents a therapeutic option that can safely and effectively reduce gradient in challenging patients with mild postoperative coarctation. Furthermore, our data suggest that aortic obstruction often coexists with ventricular diastolic dysfunction in these patients and that relief of obstruction may play a role in improvement of function. PMID- 10827388 TI - Heparin infusion prior to stenting (HIPS) trial: final results of a prospective, randomized, controlled trial evaluating the effects of local vascular delivery on intimal hyperplasia. AB - BACKGROUND: Local delivery of pharmacologic agents or genes at the site of angioplasty is a promising approach to reduce restenosis. However, there are unresolved questions concerning the safety and feasibility of local vascular delivery in clinical practice as well as the efficacy of delivered drug. To this end, the safety, feasibility, and efficacy of local delivery of heparin were evaluated in the Heparin Infusion Prior to Stenting (HIPS) trial. METHODS AND RESULTS: A total of 179 patients were enrolled in this multicenter, randomized, prospective, core laboratory-evaluated trial. Patients were randomly assigned to 5000 U heparin either administered to the coronary artery lumen or infused into the arterial wall immediately after angioplasty and before stent placement. End points included procedural events and clinical, angiographic, and intravascular ultrasound events at 6 months. Patient groups were evenly matched. There was no difference in the incidence of arterial injury, defined as an increase in arterial dissection, acute closure, or decrease in Thrombolysis In Myocardial Infarction grade blood flow in the group receiving local delivery. At follow-up there was no difference in the major adverse event rate between intraluminal (22.7%) and local groups (24.7%). There was no difference between intraluminal and local therapy in the angiographic in-stent restenosis rate (12.5%, 12.7%) or the in-stent volumetric analysis by intravascular ultrasound (IVUS) (37.19 +/- 20. 86 mm(3) vs 43.79 +/- 25.52 mm(3)). CONCLUSIONS: Local delivery of 5000 U heparin into the arterial wall before stent implantation is safe and feasible. There was not a favorable effect of locally delivered heparin on clinical, angiographic, or IVUS end points of restenosis. The use of IVUS to measure volume of intimal hyperplasia in a multicenter, core laboratory-controlled trial is feasible. PMID- 10827389 TI - Sensitivity and specificity of transesophageal echocardiography for determination of aortic valve morphology. AB - BACKGROUND: Preoperative recognition of the presence of bicuspid aortic valve can be important in the planning of procedures. Multiplane transesophageal echocardiography may allow more accurate detection of valvular morphology than does biplane transesophageal echocardiography. METHODS AND RESULTS: The studies of 710 patients who subsequently underwent valvular or aortic surgery were reviewed in a blinded fashion. The inclusion criteria were adequate short-axis view and operative note confirmation of aortic valve morphology. Six hundred eight patients were submitted to further analysis. Four hundred three patients had aortic stenosis as the primary diagnosis. Three hundred sixty patients had biplane examinations and 248 had multiplane examinations. The sensitivity and specificity of the multiplane technique in assessing aortic valve morphology (bicuspid vs tricuspid valve) was 87% and 91%, respectively. The sensitivity and specificity of the biplane technique was 66% and 56%, respectively. Whether valves were calcified or not did not result in major changes in sensitivity and specificity for either technique. CONCLUSIONS: Multiplane transesophageal echocardiography provides a more accurate assessment of preoperative aortic valve morphology than does the biplane approach in the majority of patients. PMID- 10827390 TI - Safety assessment of perflenapent emulsion for echocardiographic contrast enhancement in patients with congestive heart failure or chronic obstructive pulmonary disease. AB - BACKGROUND: Perflenapent injectable emulsion is a newer echocardiographic contrast agent that provides chamber opacification and improved endocardial border delineation. In >1000 patients tested, adverse effects have been minimal. However, previous studies have included few patients with congestive heart failure (CHF) or severe chronic obstructive pulmonary disease (COPD). Because of their reduced cardiopulmonary reserve, these patients might be at greater risk for adverse events. METHODS AND RESULTS: The safety of perflenapent emulsion was evaluated in ambulatory patients with New York Heart Association class III to IV congestive heart failure (CHF) or severe obstructive pulmonary disease (COPD). In 2 separate multicenter, phase II, randomized, double-blind, placebo-controlled, single-administration clinical trials, 146 patients in the CHF study and 134 patients in the COPD study received either perflenapent emulsion or saline placebo during an echocardiographic examination. Perflenapent emulsion was well tolerated in patients with CHF or COPD. The adverse event rates with perflenapent emulsion (15%) were similar to those with saline (11%; P =.43) in the combined groups, and similar findings were seen when analyzed by CHF or COPD groups separately. All adverse events were rated as mild, required no treatment, resolved spontaneously, and left no sequelae. Changes from baseline in vital signs, pulse oximetry, electrocardiography, and laboratory tests were similar between treatment groups. CONCLUSIONS: These data support the safety of perflenapent emulsion for contrast echocardiography in stable ambulatory patients with CHF or COPD. PMID- 10827391 TI - Effects of losartan and captopril on left ventricular volumes in elderly patients with heart failure: results of the ELITE ventricular function substudy. AB - BACKGROUND: The mechanism by which angiotensin-converting enzyme inhibitors reduce mortality rates and disease progression in patients with heart failure is likely mediated in part through prevention of adverse ventricular remodeling. This study examined the effects of the angiotensin-converting enzyme inhibitor captopril and the angiotensin II type 1 receptor antagonist losartan on ventricular volumes and function in elderly patients with heart failure and reduced left ventricular ejection fraction (< or =40%). METHODS: Patients underwent radionuclide ventriculograms (RVG) at baseline and were randomized to either captopril (n = 16) or losartan (n = 13). After 48 weeks, another RVG was obtained. Therapy was then withdrawn for at least 5 days, and the RVG was repeated while the patient was not receiving the drug. RESULTS: At 48 weeks both captopril and losartan significantly reduced left ventricular (LV) end-diastolic volume index (135 +/- 26 to 128 +/- 23 mL/m(2) for losartan, P <.05 vs baseline; 142 +/- 25 to 131 +/- 20 mL/m(2) for captopril, P <.01; mean (SD). Captopril also reduced LV end-systolic volume index (98 +/- 24 to 89 +/- 21 mL/m(2), P <.01 vs. baseline), whereas a nonsignificant trend was observed for the losartan group (97 +/- 23 to 90 +/- 16 mL/m(2), P = not significant). The between-group differences in the changes in LV volumes were not statistically significant. After drug withdrawal, LV end-diastolic volume index remained significantly lower than baseline in the captopril group (P <.01). CONCLUSIONS: Both captopril and losartan prevent LV dilation, representing adverse ventricular remodeling, previously seen with placebo treatment. Reverse remodeling was observed in the captopril group. On the basis of these results, the relative effects on LV remodeling do not provide a rationale for a survival benefit of losartan over captopril. PMID- 10827392 TI - Five-minute recording of heart rate variability in severe chronic heart failure: correlates with right ventricular function and prognostic implications. AB - BACKGROUND: In advanced chronic heart failure (CHF), correlation between heart rate variability (HRV) and parameters of disease severity is still unclear. A reduced HRV has been related to left but not to right ventricular function parameters. Moreover, the prognostic role of spectral measures is not fully defined. We sought to assess HRV by using a short electrocardiographic recording in ambulatory patients with severe CHF to investigate the relation of HRV with clinical neurohormonal and hemodynamic parameters and to determine its predictive prognostic power. METHODS AND RESULTS: HRV was obtained from 5-minute electrocardiographic recordings in 75 ambulatory patients with CHF referred for heart transplantation screening. Standard frequency-domain parameters (total power, low-frequency power, and high-frequency power) were calculated. Prognostic value of these autonomic markers and their correlation with clinical and instrumental parameters were also assessed. A low low-frequency/high-frequency ratio was an independent predictor of cardiac events (P =.015). No correlation was found between New York Heart Association class and HRV, whereas significant correlations were identified between norepinephrine plasma levels, several hemodynamic parameters, and spectral measures (P < or =.03). A reduced HRV, particularly a low-frequency power reduction (P =.000), was highly related to indexes of right ventricular dysfunction. CONCLUSIONS: The current data indicate that spectral analysis of HRV, calculated from short electrocardiographic recordings, may represent a simple but effective means contributing to risk stratification of patients with severe CHF. Autonomic information obtained from this analysis suggests that right ventricular dysfunction may be a critical element determining autonomic imbalance in patients with severe CHF. PMID- 10827393 TI - Reduction of congestive heart failure symptoms by very early fibrinolytic therapy in acute myocardial infarction: a long-term follow-up. AB - BACKGROUND: In patients with acute myocardial infarction (MI), early fibrinolytic therapy results in improved survival and preservation of ventricular function. The purpose of the study was to determine whether very early treatment also reduces the development of congestive heart failure. METHODS AND RESULTS: During the years 1984 to 1989, 358 consecutive patients with acute MI were treated with streptokinase, 161 within the first 1.5 hours from the onset of chest pain (group A) and 197 within 1.5 to 4.0 hours (group B). In 68, fibrinolysis was initiated in the prehospital setting pioneered by our group. Symptoms related to heart failure including dyspnea on exertion, fatigue, orthopnea, paroxysmal nocturnal dyspnea, nocturia, and peripheral edema, in addition to pulmonary edema events, were assessed during 5 years of follow-up. The evaluation was based on medical records and a detailed questionnaire, which was filled in by the investigators. A favorable significant effect of very early thrombolysis on the development of most of these limiting symptoms appeared 3 months after hospital discharge and persisted thereafter (P <.05). During hospitalization, pulmonary edema attacks occurred less frequently in patients from group A (23% vs 36.5%, P <.01). This difference persisted during 4 years of follow-up (13% vs 36%, P <.001). CONCLUSIONS: Our data demonstrate that very early fibrinolytic therapy results in a significant long-term reduction of congestive heart failure-related symptoms and thereby improves the quality of life in patients after MI. PMID- 10827395 TI - Quantification of mitral regurgitation in the cardiac catheterization laboratory with contrast echocardiography. AB - BACKGROUND: There is no method of quantifying the severity of mitral regurgitation (MR) from injection of tracer directly into the left ventricular (LV) cavity, a method commonly used in the cardiac catheterization laboratory. METHODS AND RESULTS: We used a previously validated mathematical model that derives regurgitant fraction (RF) from the relative tracer washout from the left atrial (LA) and LV cavities. Thirty-nine patients referred for diagnostic cardiac catheterization with clinical evidence of possible MR were included in the study. Five milliliters of a microbubble mixture was power-injected into the LV during simultaneously performed contrast echocardiography. Relative changes in background-subtracted video intensity were measured from the LV and LA, and the resultant model-derived RF was correlated with the severity of MR on cineangiography. The severity of MR ranged from 0 to 4+ on cineangiography with corresponding model-derived RF of 0 to 0.69 on contrast echocardiography. A close linear relation was noted between angiographic severity of MR and model-derived RF on contrast echocardiography (y = 0.1x + 0.03, r = 0.89, P <.001). Contrast echocardiography was more sensitive than cineangiography for detecting mild MR. CONCLUSIONS: We describe a new method of measuring the severity of MR in the cardiac catheterization laboratory. Apart from being quantitative, this method can be safely used during cardiac catheterization in patients in whom iodinated contrast agents may be potentially harmful. PMID- 10827394 TI - New insights in cardiac structural changes in patients with Fabry's disease. AB - BACKGROUND: Fabry's disease is an X-linked recessive genetic deficiency of the enzyme alpha-galactosidase leading to the pathologic intracellular deposition of neutral glycosphingolipids. Although cardiac involvement is frequent, there is controversy regarding the character of the associated left ventricular (LV) changes and the severity of valvular involvement. METHODS: Clinical evaluation (disease severity scaling, laboratory tests, and echocardiography) was performed in 13 hemizygous men (mean age 39 +/- 10 years) and 17 heterozygous women (mean age 35 +/- 19 years). RESULTS: LV hypertrophy (LVH) was frequent in subjects older than 30 years, more often in men (61%) than in women (18%, P <.001). The degree of LVH was independently associated with age and the logarithm of alpha galactosidase activity (r(2) = 0.70, P <.001). The predominant LV geometric patterns were concentric LVH and remodeling, both present in 11 subjects (36%). Three patients had an asymmetric septal hypertrophy mimicking hypertrophic cardiomyopathy. In most subjects with LVH, the systolic function was normal and severe diastolic dysfunction (restrictive pattern) was not noted. Minor structural abnormalities of the mitral valve were found in 17 subjects (57%). The aortic valve was affected in 14 patients (47%). Valvular abnormalities were frequently accompanied by regurgitation of minor to mild degree. The presence of LVH or valvular changes was associated with increased disease severity. CONCLUSIONS: Echocardiographically detectable cardiac involvement is frequent with Fabry's disease, particularly in older subjects, and more pronounced in affected hemizygous men than in heterozygous women. LVH is frequently observed but usually not associated with significant systolic or restrictive diastolic dysfunction. Concentric LVH and remodeling appear to be the major manifestations of LV structural alteration. The frequently noted valvular abnormalities were not associated with a significant degree of regurgitation. Valvular and especially LV structural changes may serve as a useful marker of disease severity. PMID- 10827397 TI - Update on familial cancer syndromes and the skin. AB - Familial cancer syndromes reflect an inherited predisposition to develop benign and malignant tumors. Clinically, the cancers occur at an earlier age and involve multiple foci of tumor formation at multiple sites. In the past 10 years, the molecular basis of many of these cancer syndromes have been unraveled with the advent of powerful genetic technologies. Entities which were hypothesized to be related on the basis of clinical features have now been shown to be linked or disparate through genetic analysis. This article reviews some of the recent advances in the clinical and molecular aspects of familial cancer syndromes that involve the skin. (J Am Acad Dermatol 2000;42:939-69.) LEARNING OBJECTIVE: After completing this article, the reader should become (1) fluent with some basic genetic principles underlying the mechanisms of cancer predisposition and positional cloning and (2) aware of the recent breakthroughs in the identification of familial cancer syndrome disease genes. PMID- 10827398 TI - Altered basic fibroblast growth factor expression in common epidermal neoplasms: examination with in situ hybridization and immunohistochemistry. AB - BACKGROUND: The fibroblast growth factor family consists of acidic fibroblast growth factor (aFGF), basic fibroblast growth factor (bFGF), and Kaposi fibroblast growth factor (kFGF). The distribution of these growth factors in skin disease has not been determined. OBJECTIVE: The purpose of this study was to determine the distribution of these growth factors in keratinocytic lesions and normal skin. METHODS: Skin sections from common disorders of keratinocytes were examined by in situ hybridization with specific probes for aFGF, bFGF, and kFGF, and immunohistochemistry. RESULTS: Of the growth factors studied, only bFGF was present in skin. bFGF messenger RNA was highly expressed in both normal epidermis and benign and malignant epithelial neoplasms. In normal skin, bFGF was expressed predominantly in a suprabasal fashion, whereas in epithelial neoplasms, homogeneous high-level expression of bFGF was observed. CONCLUSION: bFGF is the primary member of the fibroblast growth factor expressed in the skin. The source of synthesis of bFGF is keratinocytes. Immunoreactivity for bFGF appears to be primarily confined to upper layers of the epidermis in normal skin, but is expressed at all layers of the epidermis in both benign and malignant neoplastic conditions. Genetic changes that promote epithelial tumors may also promote translation of bFGF messenger RNA into protein. Specific inhibition of bFGF activity may have application in the treatment of common skin diseases. PMID- 10827399 TI - Androgenetic alopecia in heterozygous carriers of a mutation in the human hairless gene. AB - BACKGROUND: Androgenetic alopecia is considered to be genetically determined. Recently, a rare autosomal recessive form of hereditary alopecia, termed atrichia with papular lesions (APL), was found to result from mutations in the human hairless gene. OBJECTIVE: Our aim was to assess the pattern of androgenetic alopecia in heterozygous carriers of a deleterious mutation in the human hairless gene. METHODS: Healthy male second-degree relatives (n = 31) of patients affected with APL and belonging to a large consanguineous kindred were interviewed and given a Hamilton score of baldness. DNA was obtained from each subject and analyzed for the presence of a mutation in the human hairless gene known to affect this family. The age at onset and extent of baldness were compared in healthy homozygotes and heterozygous carriers of the mutation. RESULTS: Statistical analysis of the results revealed no differences in age at onset and extent of androgenetic alopecia between the two groups of subjects. CONCLUSION: The present study reports the first attempt to characterize the phenotype of heterozygous carriers of a mutation in the human hairless gene. It indicates that the presence of a deleterious mutation in one allele of the hairless gene does not affect the pattern of androgenetic hair loss. PMID- 10827400 TI - Patients with cutaneous lupus erythematosus who smoke are less responsive to antimalarial treatment. AB - BACKGROUND: Reports suggest that cigarette smoking might interfere with the effectiveness of antimalarial therapy as first-line treatment for cutaneous lupus erythematosus. Patients refractory to this treatment often must be treated with potentially more toxic regimens. OBJECTIVE: Our purpose was to examine the effects of cigarette smoking on the therapeutic response to antimalarial agents in patients with discoid lupus erythematosus (DLE) and subacute cutaneous lupus erythematosus (SCLE). METHODS: A total of 61 patients (47 DLE, 14 SCLE) were selected on the basis of the following criteria: (1) skin biopsy was consistent with cutaneous LE, (2) a smoking history was available, and (3) an adequate trial of antimalarial therapy was completed. Patients were classified as antimalarial responders or nonresponders on the basis of descriptions in their medical records. Two-by-two table analysis was performed comparing response rates in smokers versus nonsmokers. RESULTS: A significant difference (P<.0002) in the antimalarial response rate was observed for smokers (40%) versus nonsmokers (90%). CONCLUSION: These results indicate that patients with cutaneous LE who smoke are significantly less likely to respond to antimalarial therapy. PMID- 10827401 TI - Basal cell carcinoma and risk of subsequent malignancies: A cancer registry-based study in southwest England. AB - BACKGROUND: A possible link between basal cell carcinoma (BCC) and an increased subsequent risk of experiencing further noncutaneous malignancies has been suggested in previous cancer-registry and cohort studies. OBJECTIVE: The purpose of this study was to establish whether a possible link between BCC and subsequent malignancies could be confirmed in a new population in which environmental and genetic risk factors may vary from previously studied populations. METHODS: A cohort of 13,961 cancer registry-listed persons from the southwest of England, in whom BCC had been diagnosed during the period of 1981 to 1988, was examined for the relative risk of experiencing various further malignancies. RESULTS: An approximately 3-fold increase in the risk for malignant melanoma was demonstrated. No other cancers occurred in statistically significant excess. CONCLUSION: The previous reported associations of BCC onset with subsequent increased risk for various noncutaneous cancers are not supported by this study. PMID- 10827402 TI - Parastomal pyoderma gangrenosum: clinical features and management. AB - BACKGROUND: The importance of pyoderma gangrenosum (PG) as a cause of ulceration around abdominal stomas is not well recognized. OBJECTIVE: Our purpose was to describe the incidence, clinical and histologic features, disease associations, and possible risk factors for parastomal PG. METHODS: A clinic, run by a dermatologist and two stoma nurses, was created. Five hundred patients approached by postal questionnaire were invited to attend if they had skin problems. In addition, local surgical, dermatologic, and nursing services were invited to refer patients with parastomal skin problems. Cases of parastomal PG were identified, investigated, and treated. RESULTS: The annual incidence of parastomal PG in the questionnaire-based cohort of patients was 0.6% (3 patients). An additional 23 patients with the condition were seen. No consistent hematologic, biochemical, immunologic, microbiological or histologic abnormalities were identified. Local skin damage did not appear to be an important trigger for parastomal PG. The condition is recurrent in one third of cases. Topical tacrolimus (0.3% in carmellose sodium paste) has been effective in 4 patients. CONCLUSION: Parastomal PG is far more common than previous reports would suggest, and it may be associated with diseases other than inflammatory bowel disease. PMID- 10827403 TI - Vertex balding, plasma insulin-like growth factor 1, and insulin-like growth factor binding protein 3. AB - BACKGROUND: A recent report suggested that men with vertex balding have higher levels of plasma insulin-like growth factor 1 (IGF-1). The association of its major carrier protein, insulin-like growth factor binding protein 3 (IGFBP-3), with male pattern hair loss has not been examined. OBJECTIVE: We evaluated the relations of plasma concentrations of IGF-1 and IGFBP-3 with vertex balding in middle-aged and elderly men. METHODS: Participants were 431 male members of the Health Professionals Follow-up Study who responded to a question in 1992 on their hair pattern at 45 years of age and who were 47 to 81 years old when they provided a blood specimen in 1993-1994. Odds ratios (ORs) of vertex balding associated with IGF-1 and IGFBP-3 were estimated from logistic regression models mutually adjusting for each other and controlling for age at blood draw. RESULTS: Of the 431 men, 128 had vertex balding at age 45. Compared with men who were not balding, for a 1 standard deviation increase in plasma IGF-1 level (72.4 ng/mL), the OR for vertex balding was 1. 31 (95% CI, 0.95-1.81). For a 1 standard deviation increase in plasma IGFBP-3 (957 ng/mL), the OR for vertex balding was 0.62 (95% CI, 0.44-0.88). CONCLUSION: Older men with vertex balding have lower circulating levels of IGFBP-3 and higher levels of IGF-1 when controlling for IGFBP-3 level. PMID- 10827404 TI - In vitro compatibility of tazarotene with other topical treatments of psoriasis. AB - Tazarotene is the first receptor-selective retinoid indicated for the topical treatment of plaque psoriasis. It is being used clinically in combination with other topical antipsoriatic treatments, although its stability in the presence of these products has not been examined extensively. This study examines the compatibility of tazarotene 0.05% gel with 17 other topical products used in the treatment of psoriasis, assessed over a 2-week period. Tazarotene showed minimal degradation (<10%) at 0, 8, 24, and 48 hours after compounding with each of the 17 products. In addition, after 1 and 2 weeks, degradation of tazarotene remained less than 10% for 15 of the 17 products tested. Tazarotene appeared to have minimal impact on the stability of the other products. These results suggest that tazarotene gel can be successfully coprescribed with a range of commonly used topical psoriasis treatments without adversely affecting the chemical stability of either agent. PMID- 10827406 TI - Topical corticosteroid in foam vehicle offers comparable coverage compared with traditional vehicles. AB - BACKGROUND: A new preparation of betamethasone valerate in a novel foam vehicle is available for treatment of scalp dermatoses. The vehicle spreads between hair until it reaches the scalp, where it melts and delivers the active drug. Solids make up only a tiny fraction of the foam vehicle, leaving no apparent residue on the skin or hair. The uniqueness of this vehicle raises the question of how to compare it with other topical corticosteroid preparations. OBJECTIVE: The purpose of this study was to determine the equivalency of a given quantity of the foam product to quantities of drugs in conventional vehicles. METHODS: The number of fingertip units (FTUs) per gram and the area of coverage of an FTU of betamethasone valerate foam vehicle were determined and compared with those of cream, lotion, gel, and solution psoriasis treatments. RESULTS: The weight of an FTU of foam vehicle was 52.5 +/- 5.7 microg. There were 9 to 12 times as many FTUs in 100 g of vehicle foam as in 100 g of cream or gel and 2.3 to 2.8 times as many as in 100 g of lotion or solution. The area covered by an FTU of foam vehicle was less than the area covered by an FTU of cream or gel. CONCLUSION: The characteristics of foam vehicle are different from those of other vehicles. The greater number of FTUs in 100 g of foam vehicle made up for the lower coverage per FTU, such that total coverage area for 100 g of foam vehicle was comparable to the coverage area for 100 g of the drugs in traditional vehicles. PMID- 10827405 TI - Lobular capillary hemangiomas: An epidemiologic report, with emphasis on cutaneous lesions. AB - BACKGROUND: Lobular capillary hemangiomas (pyogenic granulomas) occur on both mucosal and cutaneous surfaces. There are conflicting data regarding the increased prevalence of lobular capillary hemangiomas in female versus male subjects. Some studies have noted a female predominance of lobular capillary hemangiomas, but other studies do not reveal such a disparity. Because of an increased prevalence during pregnancy, oral tumors are also known as "granuloma gravidarum" or "pregnancy tumors." A hormonal influence for these mucosal lesions has been postulated. There are, however, no studies that address a possible relationship between hormones and cutaneous lesions. OBJECTIVE: This study presents the epidemiology of lobular capillary hemangiomas, with an emphasis on cutaneous lesions. METHODS: We reviewed 63,759 dermatopathology reports from a regional, private dermatopathology laboratory and found 325 cases of lobular capillary hemangiomas over a 1-year period. RESULTS: In our study of lobular capillary hemangiomas, cutaneous lesions accounted for 86%, with mucosal lesions representing only 12% of cases. Seven cases were excluded (one was intravascular, two were subcutaneous, and in 4 the location was not specified). Overall, male patients outnumbered female patients. The peak incidence for cutaneous lobular capillary hemangiomas was found in the second decade of life. The most common cutaneous sites were the trunk, upper extremities, and head. Mucosal lesions were primarily seen on the lips, gingiva, and tongue, and these affected females more than males by a ratio of 2:1, most commonly in the fourth decade of life. CONCLUSION: Cutaneous lobular capillary hemangiomas were equally prevalent in male and female patients. This would refute a female hormonal influence in the induction of cutaneous lobular capillary hemangiomas. Our data may suggest a hormonal influence on mucosal lesions because mucosal lobular capillary hemangiomas were twice as common in female patients. However, the small number of lesions in our study precludes us from making such a conclusion. PMID- 10827407 TI - Exacerbation of pustulosis palmaris et plantaris after topical application of metals accompanied by elevated levels of leukotriene B4 in pustules. AB - BACKGROUND: Pustulosis palmaris et plantaris (PPP) is a chronic inflammatory disease consisting of polymorphonuclear leukocyte infiltration, and is often exacerbated by focal infections such as tonsillitis. In some cases, metal allergy has been reported. OBJECTIVE: The purpose of this study was to evaluate (1) the significance of metal allergy in the formation of pustules, and (2) the participation of leukotriene (LT) B(4) in the formation of pustules of PPP. METHODS: Patch tests with metals were performed on 7 patients with PPP, and both pustular and plasma levels of LTB(4) were measured in these 7 patients before and 48 hours after metal patch tests. RESULTS: Palmoplantar pustules were exacerbated after the metal patch tests in all 7 patients. The mean levels of LTB(4) in plasma and pustules of the volar surface at 48 hours after the metal patch tests were significantly higher than those before the metal patch tests. CONCLUSION: Metals can be important in the pathogenesis of PPP by contributing to the induction of high LTB(4) concentration in the pustules. PMID- 10827408 TI - Botulinum toxin type A therapy for palmar and digital hyperhidrosis. AB - OBJECTIVE: We evaluated the efficacy of subepidermal injections of botulinum toxin type A on recalcitrant palmar and digital hyperhidrosis. METHODS: Twenty patients with recalcitrant palmar and digital hyperhidrosis were treated with subepidermal injections of botulinum toxin. Nineteen patients completed the 12 month study. Injections were performed in 3 stages. The total dose of toxin for each hand, which included the palm, thenar eminence, and digits, was 165 units. Patients were followed up on a monthly basis. RESULTS: Botulinum toxin significantly reduced sweat production in the treated areas. Anhidrosis lasted 9 months in 3 patients, 8 months in 3 patients, 7 months in 8 patients, 6 months in 3 patients, 5 months in 1 patient, and 4 months in 1 patient. Reduced sweating of the palm and digits continued in all patients for the 12-month evaluation period, with the greatest reduction of sweating in the nondominant hand. Mild weakness of the thumb occurred in 4 patients at a mean duration of 3 weeks, with the greatest duration being 6 weeks. CONCLUSION: Botulinum toxin provides a safe and efficacious alternative in the treatment of recalcitrant palmar and digital hyperhidrosis. PMID- 10827409 TI - UVA rush hardening for the treatment of solar urticaria. AB - Induction of tolerance by subsequent UV exposures is the most effective therapy for solar urticaria; however, it is time-consuming and takes a long time until protection is achieved. Three patients with solar urticaria were exposed to multiple UVA irradiations at 1-hour intervals per day. With this rush hardening regimen, protection was achieved within 3 days. PMID- 10827410 TI - Clinical improvement and immunohistochemical findings in severe atopic dermatitis treated with interferon gamma. AB - BACKGROUND: Several clinical studies have focused on the therapeutic effects of interferon gamma (IFN-gamma) in patients with severe atopic dermatitis (AD), although the dosage of recombinant IFN-gamma (rIFN-gamma), therapeutic schedule, and the degree of clinical improvement were different among studies. Although the exact mechanism of action of IFN-gamma therapy in AD is not clear, the beneficial effects of IFN-gamma have been attributed mainly to an immunomodulating effect on the expression of certain immunologic markers. OBJECTIVE: Our purpose was to study the therapeutic effect of two different dosages of rIFN-gamma on AD and to investigate the change of lesional expression of infiltrating inflammatory cell markers associated with rIFN-gamma therapeutic efficacy. METHODS: Fifty-one patients with severe recalcitrant AD were treated with rIFN-gamma. Twenty patients were treated with 0.5 x 10(6) IU/m(2) of rIFN-gamma (low-dose [LD] group); 21 patients received 1.5 x 10(6) IU/m(2) of rIFN-gamma (high-dose [HD] group); and 10 patients received placebo. The patients were injected subcutaneously 3 times a week for 12 weeks. Immunohistochemical study was performed in 20 patients of the HD group in the initial visit and after completion of rIFN-gamma therapy with a panel of 14 monoclonal antibodies as markers of inflammatory cells and cytokines. RESULTS: The disease severity of the 2 groups treated with rIFN-gamma was reduced significantly at the end of treatment compared with that of the placebo group (P<.05). More rapid clinical improvement and more effective treatment outcome were seen in the HD group than in the LD group for the initial 6-week treatment period; however, the clinical improvement in both of the treated groups was stable and maintained after week 8 of treatment. Immunohistochemical findings showed statistically significant reduction in the lesional expression of CD25 and EG2 cells that infiltrated into skin after rIFN-gamma therapy. CONCLUSION: This study demonstrated that rIFN gamma therapy for AD is safe and effective. In the early phase of therapy, a higher dosage of rIFN-gamma is more effective; and for the maintenance of clinical improvement, a lower dosage of rIFN-gamma is recommended when high cost and effectiveness of rIFN-gamma are considered. The therapeutic efficacy of rIFN gamma in AD might be in part related to the decreased number of CD25(+) and EG2(+) inflammatory cells infiltrated into skin. PMID- 10827411 TI - The use of full-thickness skin grafts to repair nonperforating nasal defects. AB - BACKGROUND: Full-thickness skin grafts are traditionally thought of as inferior alternatives to local flaps in nasal reconstruction. OBJECTIVE: Our purpose was to demonstrate that full-thickness skin grafts are viable alternatives to local flaps in the reconstruction of nonperforating defects extending through the dermis of the nose. METHODS: The literature was reviewed to determine the best donor sites for full-thickness skin grafts to repair defects on different regions of the nose. RESULTS: The optimal donor site for the repair of defects on the thin skin of the upper two thirds of the nose (dorsum and sidewall) is the preauricular area. The best donor site for the repair of defects on the thick, sebaceous skin of the lower one third of the nose (tip and ala) is the conchal bowl of the ear. Defects located on the transition zone between the upper two thirds and lower one third of the nose are best repaired with Burow's grafts or nasolabial fold grafts. CONCLUSION: Full-thickness skin grafts are excellent alternatives to local flaps in the reconstruction of nonperforating nasal defects provided that the surgeon selects the appropriate donor site based on whether the surgical defect is located on the thin, pliable skin of the upper nose or the thick, sebaceous skin of the lower nose. PMID- 10827412 TI - Revised classification system for inherited epidermolysis bullosa: Report of the Second International Consensus Meeting on diagnosis and classification of epidermolysis bullosa. PMID- 10827413 TI - Double nicking of Mohs tissue specimen. PMID- 10827414 TI - Annular erythema of Sjogren's syndrome in a white woman. AB - Annular erythema of Sjogren's syndrome (AE-SS) is believed to be the Asian counterpart of subacute cutaneous lupus erythematosus (SCLE) in white persons. We report the second case of a white person with AE-SS, diagnosed by clinical and serologic findings, as well as the absence of histologic criteria for SCLE. The diagnosis of AE-SS was established by symptoms of xerophthalmia and xerostomia, as well as by examination of skin and salivary gland biopsy specimens. Evaluation showed the presence of anti-Ro(SS-A) and anti-La(SS-B) autoantibody, with the presence of anti-Ro(SS-A) antibody against the 60-kd, but not the 52-kd, epitope, a pattern frequently seen in both the Asians with AE-SS and in white patients with SCLE. Both skin and sicca symptoms were alleviated with combination antimalarial therapy, which included hydroxychloroquine and quinacrine. This case demonstrates that AE-SS can occur in white patients and that the autoantibody profile is similar to that described in Asians with this disease. PMID- 10827415 TI - Computer palms. AB - We describe a new occupation-related skin finding in 2 computer programmers and discuss its characteristics and causes. PMID- 10827416 TI - Migratory poliosis: A forme fruste of alopecia areata? AB - We describe a 19-year-old African-American man with a 14-year history of migratory poliosis. We suggest that this phenomenon may represent a forme fruste of alopecia areata. PMID- 10827417 TI - Vulvitis circumscripta plasmacellularis mimicking child abuse. AB - Vulvitis circumscripta plasmacellularis (VCP) is a rare, benign vulvar disorder that is typically described in adult women. Our case occurred in an 8-year-old girl. The primary diagnostic concern was sexual abuse. VCP may also mimic lichen sclerosus, extramammary Paget's disease, pemphigus vulgaris, fixed drug eruption, squamous cell carcinoma, candidiasis, allergic contact dermatitis, and herpes simplex infection. Evaluation should include a biopsy because the histopathologic features of VCP are distinctive. PMID- 10827418 TI - The dermatologist, genetic counseling, and cancer-associated genodermatoses. PMID- 10827419 TI - The foul side of fragrance-free products. PMID- 10827420 TI - Evidence is lacking for a synergistic or additive effect of combination extracorporeal photopheresis with interferon alfa for cutaneous T-cell lymphoma. PMID- 10827422 TI - Out of Africa: training collaboration and malaria research. PMID- 10827423 TI - Plant-based malaria control: research initiative on traditional antimalarial methods. PMID- 10827424 TI - Structure-function analysis of malaria proteins by gene targeting. PMID- 10827426 TI - Passports, tiny worms and herbal teas On the Web PMID- 10827425 TI - Structure-function analysis of malaria proteins by gene targeting--a response. PMID- 10827427 TI - Abnormal blood flow and red blood cell deformability in severe malaria. AB - Obstruction of the microcirculation plays a central role in the pathophysiology of severe malaria. Here, Arjen Dondorp and colleagues describe the various contributors to impaired microcirculatory flow in falciparum malaria: sequestration, rosetting and recent findings regarding impaired red blood cell deformability. The correlation with clinical findings and possible therapeutic consequences are discussed. PMID- 10827428 TI - Hormones and nuclear receptors in schistosome development. AB - A substantial but disparate body of evidence suggests that hormones affect the development of schistosomes within their definitive hosts. Here, Raymond Pierce and colleagues review such evidence for host steroid and thyroid hormones, and for ecdysteroids, and link this to the expanding knowledge of the nuclear receptors for these hormones. Phylogenetic analysis of the nuclear receptor superfamily and the characterization of the first schistosome nuclear receptors suggest that steroids and thyroid hormone probably act indirectly, or by pathways not involving the control of gene transcription. However, the probability that schistosome nuclear receptors exist for a variety of unique ligands opens up exciting possibilities for targeted drug development. PMID- 10827429 TI - Motile systems in malaria merozoites: how is the red blood cell invaded? AB - The ability of the malaria parasite to invade erythrocytes is central to the disease process, but is not thoroughly understood. In particular, little attention has been paid to the motor systems driving invasion. Here, Jennifer Pinder, Ruth Fowler and colleagues review motility in the merozoite. The components of an actomyosin motor are present, including a novel unconventional class XIV myosin, now called Pfmyo-A, which, because of its time of synthesis and location, is likely to generate the force required for invasion. In addition, there is a subpellicular microtubule assemblage in falciparum merozoites, the f MAST, the integrity of which is necessary for invasion. PMID- 10827430 TI - Evolutionarily conserved proteins as prominent immunogens during Leishmania infections. AB - Many Leishmania antigens have been identified as members of conserved protein families, such as the acidic ribosomal proteins, the histones and the heat-shock proteins; despite this, they elicit specific immune responses. Furthermore, homologues of many of these antigens are immune targets in other infectious diseases and systemic autoimmune diseases. Here, Jose Mar a Requena, Carlos Alonso and Manuel Soto review this class of widely distributed antigens, which they call 'panantigens'. They also propose a model to explain the prominent immunogenicity of these antigens during Leishmania infection, on the basis of the fact that many panantigens are constituents of multicomponent complexes in the cell. The elucidation of the pathways by which Leishmania antigens are processed and presented to effector cells from the host immune system will shed light on the immunopathology of leishmaniasis and help in the development of protective immunotherapies. PMID- 10827431 TI - Websites of interest PMID- 10827433 TI - Corrigendum PMID- 10827432 TI - The economic burden of lymphatic filariasis in India. AB - Lymphatic filariasis affects 119 million people living in 73 countries, with India accounting for 40% of the global prevalence of infection. Despite its debilitating effects, lymphatic filariasis is given very low control priority. One of the reasons for this is paucity of information on the economic burden of the disease. Recent studies in rural areas of south India have shown that the treatment costs and loss of work time due to the disease are considerable. Based on the results of these studies, Kapa Ramaiah et al. here estimate the annual economic loss because of lymphatic filariasis for India and discuss the implications of their findings. PMID- 10827434 TI - Cotrimoxazole for prenatal treatment of congenital toxoplasmosis? AB - Congenital toxoplasmosis is still one of the most frequent causes of fetal death. Despite a significant improvement in diagnosis, particularly in utero diagnosis, maternal treatment is only partially effective in preventing transmission to the fetus and treating fetal infection. Maternal treatment is based on drugs developed 50 years ago, which may have limited efficacy (spiramycin) or serious side-effects (pyrimethamine). Data on the use of cotrimoxazole in mouse models of toxoplasmosis and for preventing toxoplasmic encephalitis in patients suffering from AIDS have led Francis Derouin and colleagues to consider the potential of cotrimoxazole for prenatal prevention and treatment of toxoplasmic fetal death. PMID- 10827435 TI - Schistosoma haematobium in Morocco: moving from control to elimination. AB - In this article, Hammou Laamrani and colleagues summarize the Moroccan schistosomiasis control programme and discuss the challenges ahead for schistosomiasis elimination. In 1994, a programme was initiated by the Moroccan Ministry of Health to eliminate schistosomiasis from Morocco by the year 2004. In 1997, this objective had been achieved in three out of 20 affected provinces. This article discusses the background and strategies of this programme, as well as the achievements, the problems encountered and the challenges ahead, along with suggestions as to how to reach the goal of elimination of urinary schistosomiasis in Morocco, and possibly elsewhere in Africa. PMID- 10827436 TI - New data from old Trypanosomatid preparations. AB - Assessing the diversity of the Trypanosomatidae is difficult because of the relatively small number of species that can be cultured. This same problem thwarts efforts to identify the hosts and insect vectors of Phytomonas, a genus of parasites of plants that includes species responsible for devastating epiphytotics of economically important plantations. Here, Myrna Serrano, Marta Teixeira and Erney Camargo review the studies that have led to the development of a PCR-based technique for processing insect and plant juices fixed on glass slides. The method overcomes the need for cultivation, facilitates field collections and also permits the molecular examination of archival smears of Phytomonas. In principle, the method can be adapted to any trypanosomatid as well as to any fastidious parasitic or free-living organism. PMID- 10827437 TI - More on Babesia, NO and malaria. PMID- 10827438 TI - Reply PMID- 10827439 TI - Parasite candidate vaccines: a warning from polymorphic Leishmania populations. PMID- 10827440 TI - Reply PMID- 10827441 TI - Is horsemeat trichinellosis an emerging disease in the EU? PMID- 10827442 TI - Controversy about the visual magnocellular deficit in developmental dyslexics. PMID- 10827443 TI - On the conflicting support for the magnocellular-deficit theory of dyslexiaResponse to Stein, Talcott and Walsh (2000). PMID- 10827444 TI - Cognitive and emotional influences in anterior cingulate cortex. AB - Anterior cingulate cortex (ACC) is a part of the brain's limbic system. Classically, this region has been related to affect, on the basis of lesion studies in humans and in animals. In the late 1980s, neuroimaging research indicated that ACC was active in many studies of cognition. The findings from EEG studies of a focal area of negativity in scalp electrodes following an error response led to the idea that ACC might be the brain's error detection and correction device. In this article, these various findings are reviewed in relation to the idea that ACC is a part of a circuit involved in a form of attention that serves to regulate both cognitive and emotional processing. Neuroimaging studies showing that separate areas of ACC are involved in cognition and emotion are discussed and related to results showing that the error negativity is influenced by affect and motivation. In addition, the development of the emotional and cognitive roles of ACC are discussed, and how the success of this regulation in controlling responses might be correlated with cingulate size. Finally, some theories are considered about how the different subdivisions of ACC might interact with other cortical structures as a part of the circuits involved in the regulation of mental and emotional activity. PMID- 10827445 TI - The distributed human neural system for face perception. AB - Face perception, perhaps the most highly developed visual skill in humans, is mediated by a distributed neural system in humans that is comprised of multiple, bilateral regions. We propose a model for the organization of this system that emphasizes a distinction between the representation of invariant and changeable aspects of faces. The representation of invariant aspects of faces underlies the recognition of individuals, whereas the representation of changeable aspects of faces, such as eye gaze, expression, and lip movement, underlies the perception of information that facilitates social communication. The model is also hierarchical insofar as it is divided into a core system and an extended system. The core system is comprised of occipitotemporal regions in extrastriate visual cortex that mediate the visual analysis of faces. In the core system, the representation of invariant aspects is mediated more by the face-responsive region in the fusiform gyrus, whereas the representation of changeable aspects is mediated more by the face-responsive region in the superior temporal sulcus. The extended system is comprised of regions from neural systems for other cognitive functions that can be recruited to act in concert with the regions in the core system to extract meaning from faces. PMID- 10827446 TI - The complementary brain: unifying brain dynamics and modularity. AB - How are our brains functionally organized to achieve adaptive behavior in a changing world? This article presents one alternative to the computer analogy that suggests brains are organized into independent modules. Evidence is reviewed that brains are in fact organized into parallel processing streams with complementary properties. Hierarchical interactions within each stream and parallel interactions between streams create coherent behavioral representations that overcome the complementary deficiencies of each stream and support unitary conscious experiences. This perspective suggests how brain design reflects the organization of the physical world with which brains interact. Examples from perception, learning, cognition and action are described, and theoretical concepts and mechanisms by which complementarity might be accomplished are presented. PMID- 10827447 TI - Positioning the isthmic organizer where Otx2 and Gbx2meet. AB - Regional diversity along the anterior-posterior axis of the central nervous system is established during gastrulation and is subsequently refined by local organizing centres that are located at genetically defined positions. The isthmic organizer possesses midbrain- and cerebellum-inducing properties, and its positioning at the midbrain-hindbrain boundary is a crucial event that controls midbrain and cerebellum development. Recent work has shown that two transcription factors, Otx2 and Gbx2, are instrumental in positioning the isthmic organizer at the midbrain-hindbrain boundary. PMID- 10827448 TI - Segregation distortion of mouse t haplotypes the molecular basis emerges. AB - The t haplotype is an ancestral version of proximal mouse chromosome 17 that has evolved mechanisms to persist as an intact genomic variant in mouse populations. t haplotypes contain mutations that affect embryonic development, male fertility and male transmission ratio distortion (TRD). Collectively, these mutations drive the evolutionary success of t haplotypes, a phenomenon that remains one of the longstanding mysteries of mouse genetics. Molecular genetic analysis of TRD has been confounded by inversions that arose to lock together the various elements of this complex trait. Our first molecular glimpse of the TRD mechanism has finally been revealed with the cloning of the t complex responder (Tcr) locus, a chimeric kinase with a genetically cis active effect. Whereas + sperm in a +/t male have impaired flagellar function caused by the deleterious action of trans-active, t haplotype-encoded 'distorters,' the mutant activity of Tcr counterbalances the distorter effects, maintaining the motility and fertilizing ability of t sperm. PMID- 10827449 TI - The eye: monophyletic, polyphyletic or perhaps biphyletic? PMID- 10827450 TI - Reply to meyer-rochow PMID- 10827451 TI - Sex in the wormcounting and compensating X-chromosome dose. AB - The nematode Caenorhabditis elegans counts its X chromosomes to determine sex and to activate the process of dosage compensation, which ensures that males (XO) and hermaphrodites (XX) express equal levels of most X-chromosome products. The number of X chromosomes is communicated by a set of X-linked genes called X signal elements, which repress the master sex-determination switch gene xol-1 via two distinct, dose-dependent molecular mechanisms in XX embryos. X-chromosome gene dosage is compensated by a specialized protein complex that includes evolutionarily conserved components of mitotic and meiotic machinery. This complex assembles on both X chromosomes of hermaphrodites to repress transcription by half. The recruitment of chromosome segregation proteins to the new task of regulating X-chromosome-wide gene expression points to the evolutionary origin of nematode dosage compensation. PMID- 10827452 TI - Forward with BACs: new tools for herpesvirus genomics. AB - The large, complex genomes of herpesviruses document the high degree of adaptation of these viruses to their hosts. Not surprisingly, the methods developed over the past 30 years to analyse herpesvirus genomes have paralleled those used to investigate the genetics of eukaryotic cells. The recent use of bacterial artificial chromosome (BAC) technology in herpesvirus genetics has made their genomes accessible to the tools of bacterial genetics. This has opened up new avenues for reverse and forward genetics of this virus family in basic research, and also for vector and vaccine development. PMID- 10827453 TI - Partners and pathwaysrepairing a double-strand break. AB - Double-strand chromosome breaks can arise in a number of ways, by ionizing radiation, by spontaneous chromosome breaks during DNA replication, or by the programmed action of endonucleases, such as in meiosis. Broken chromosomes can be repaired either by one of several homologous recombination mechanisms, or by a number of nonhomologous repair processes. Many of these pathways compete actively for the repair of a double-strand break. Which of these repair pathways is used appears to be regulated developmentally, genetically and during the cell cycle. PMID- 10827454 TI - RTK mutations and human syndromeswhen good receptors turn bad. AB - Mutations in receptor tyrosine kinases (RTKs) have been linked to an increasing number of inherited human disease syndromes, including dwarfism, craniosynostosis, heritable cancer susceptibility, venous malformation and Piebaldism. Both gain-of-function mutations resulting in constitutive receptor activation, and loss-of-function mutations resulting in non-functional or dominant negative receptors, have been observed. This review summarizes RTK families that are involved in inherited syndromes, describes the molecular consequences of the disease mutations, and predicts that many novel mutations remain to be identified. PMID- 10827455 TI - US gene therapy in crisis. AB - The recent death of a young man, Jesse Gelsinger, in a gene therapy experiment has sent shock waves throughout the US research community. This tragic event has raised new questions about the prospects for human gene therapy. And it has also reopened old controversies about how best to guide the development of this field. PMID- 10827456 TI - EMBOSS: the European Molecular Biology Open Software Suite. PMID- 10827458 TI - [Adhesive properties of basal membranes of Bruch's membrane. Immunohistochemical studies of age-dependent changes in adhesive molecules and lipid deposits]. AB - BACKGROUND: In addition to deposition of debris between the plasma and basal membrane of the RPE, different types of early and late AMD-like drusen, choroidal neovascularization and pigment epithelial detachments are characterized by splitting of Bruch's membrane between the basement membrane of the RPE and the inner collagenous layer. Because these layers are normally attached by adhesion molecules, the possible age-related regression of these adhesion molecules in this structure was examined. METHODS: The presence of the adhesion molecules laminin, fibronectin, collagen IV and of lipid deposits was examined in 44 macular specimen (age 4-88 years). RESULTS: Age-related regression of the adhesion molecules laminin, fibronectin and collagen IV was observed in the basement membrane of the RPE and the endothelium cells of the choriocapillaris. This effect correlated with an increase in lipid deposits. CONCLUSION: The regression of adhesion molecules, especially in the basement membrane of the RPE in association with increased lipid deposits in Bruch's membrane, may explain the development of a "cleavage edge" for the splitting of Bruch's membrane between the basement membrane of the RPE and the inner collagenous layer. PMID- 10827459 TI - [Findings of optical coherence tomography (OPT) before and after macular hole surgery]. AB - PURPOSE: We used optical coherence tomography (OCT) for the staging of macular holes and compared our results with clinical and intraoperative findings. PATIENTS: Between 1 January 1997 and 15 October 1998, 80 patients underwent vitrectomy for idiopathic macular holes. In 36 cases we could record an OCT preoperatively and in 24 cases also postoperatively. OCT measurements were analyzed in a blind fashion independent of the clinical pre- and intraoperative findings. RESULTS: Preoperative OCT staging was in agreement with the intraoperative findings in 24 of 36 cases. However, in 8 cases, the macular hole was judged as stage 3 intraoperatively, while OCT revealed a stage 4 hole. Postoperatively, OCT results were consistent with the clinical findings in all 24 cases. CONCLUSION: OCT scans are a valuable adjunct for the pre- and postoperative analysis of idiopathic macular holes. As the posterior hyaloid surface cannot always be reliably identified in OCT, discrimination between stage 3 and 4 is difficult. PMID- 10827460 TI - [Endophthalmitis after cataract surgery: predisposing factors, infectious agents and therapy]. AB - BACKGROUND: Infectious exogenous endophthalmitis is a serious complication after cataract surgery. Despite modern pharmacological and surgical methods, its treatment is still difficult. PATIENTS AND METHODS: In a retrospective study the records of all patients treated for endophthalmitis following cataract extraction at the Department of Ophthalmology of the University Hospital in Hamburg between January 1989 and December 1997 were assessed. RESULTS: Of 36 patients treated for endophthalmitis, 29 (80.6%) had been referred. In 14 (38.9%) of these 29 patients endophthalmitis had occurred after outpatient cataract surgery. Seven patients (19.4%) had been treated as inpatients at the University Eye Hospital Eppendorf. Vitrectomy was performed in 80.6% of the cases. An infectious agent was isolated from 50% of diagnostic probes. The most common organism isolated were coagulase negative staphylococci (4 cases). Predisposing factors for the development of endophthalmitis were diabetes (27.8%), intraoperative loss of vitreous (19.4%), application of systemic steroids (13.9%) and wound dehiscence (11.1%). Of 27 patients (75%) followed up, 16 (59.3%) had a final visual acuity of 20/400 or better (mean 20/40). An enucleation had to be performed in 4 patients (13.8%). CONCLUSION: In this study, approximately 60% of patients with endophthalmitis following cataract extraction had their globes preserved and a good visual outcome after appropriate surgical interventions. PMID- 10827461 TI - [Pupil physiology after cataract surgery]. AB - BACKGROUND: The goal of a cataract operation is to achieve an optimal outcome, which includes a round and functioning pupil. The goal of this study was to analyze the influence of cataract operations on pupils that appear to be normal on slit-lamp examination postoperatively. PATIENTS AND METHODS: A videopupillography was performed on 47 eyes of 47 patients after phacoemulsification without complications in the first eye, and on 12 eyes of 12 patients after phacoemulsification without complications in the second eye. RESULTS: After surgery, the first eyes showed a significantly more constricted pupil compared to the non-operated eye independently of the stage of irritation. After surgery of the second eye, this was only observed under near dark conditions. CONCLUSION: Even the morphologically normal pupil shows a change in motility postoperatively. We believe that the reason for this is the mechanical manipulation of the iris during cataract operation. During this, the sensitive dilator muscle of the pupil is affected, leading to temporary or permanent changes in pupillary function. The change in motility of the contralateral, non operated eye cannot be easily explained. The pupil size is important for the visual process and alteration of its function allows an evaluation of the quality of the operation. PMID- 10827462 TI - [Ultrasound biomicroscopy in pigmentary glaucoma]. AB - PURPOSE: To evaluate the anatomical relationships of the iris in pigmentary glaucoma before and after laser iridotomy and to evaluate the effect on intraocular pressure. METHODS: Ultrasound biomicroscopy (UBM, Humphrey-Zeiss) of the anterior segment was performed in 28 eyes of 28 patients (20 male, 8 female, mean age 43 years, mean untreated IOP 24.3 mmHg) with pigmentary glaucoma before and after laser iridotomy. The slope of intraocular pressure was documented. Mean follow-up was 9 months. For statistical analysis the Wilcoxon test was used. RESULTS: Ten out of 28 eyes showed iridozonular contact and concavity of the midperipheral iris. Laser iridotomy resulted in a significant pressure drop (P < 0.05) in these 10 eyes (24.6 mmHg to 18.3 mmHg). Eighteen eyes, however, did not show iridozonular contact and intraocular pressure did not drop sufficiently (P > 0.05; 25.1 mmHg to 23.1 mmHg) after laser iridotomy. CONCLUSION: The results show that iridozonular contact does not exist in every patient with pigmentary glaucoma. Therefore, it seems possible that more than one pathogenic mechanism is involved in pigmentary glaucoma. In patients with iridozonular contact, however, laser iridotomy significantly reduces intraocular pressure. PMID- 10827463 TI - [Imaging of orbital metastasis of small-intestine carcinoid by octreotide scintigraphy. A critical evaluation of the methodology]. AB - BACKGROUND: An orbital carcinoid metastasis can be specifically imaged by octreotide scintigraphy. Orbital metastases of carcinoid tumors are rare. In the current literature only 27 cases have been published. CASE REPORT: We report on a 59 year-old woman who presented with diplopia 6 years after resection of a carcinoid tumor of the ileum. She also complained about flush and diarrhea. The diplopia was caused by limited extensibility of the right inferior muscle. Computed tomography with contrast revealed an enhancing tumor in the area of the right inferior rectus muscle, which appeared in the MRT as an isointense structure in the T1-weighted image. On staging, we found a contrast-enhancing nodular hepatic lesion. In octreotide scintigraphy, enhancement of the liver, thoracic and abdominal para-aortic lymph nodes and of the right inferior orbit was found. CONCLUSION: The survival times of patients with orbital metastasis of carcinoid tumors range between 7 months and 11 years. No specific treatment is available. Surgical excision is indicated for a symptomatic solitary metastasis. Octreotide is helpful in two ways: (1) as a radiolabeled tracer to detect extra abdominal and extrahepatic secondary tumors; (2) as a treatment modality for flush and diarrhea, which is successful in over 60% of patients (life expectancy uninfluenced). Our patient is presently being treated with 200 micrograms of octreotide three times a day subcutaneously. Flush episodes and diarrhea have been reduced, but her ophthalmologic symptoms remain unchanged, and she has recently developed cardiac insufficiency. PMID- 10827464 TI - [Role of vitreoretinal interface in the pathogenesis and therapy of macular disease associated with optic pits]. AB - PURPOSE: Although the relationship between optic pits and macular lesions was described nearly a century ago, the pathology and pathogenesis of macular detachment remain unclear. Recent OCT studies have shown schisislike spaces in connection with the disc. None of the hypotheses of pathogenesis proposed so far could have been proven. Besides the hypothesis of exudation, the role of the vitreoretinal interface is kept in the background of discussion. We describe a case of macular detachment associated with optic pit that regained full vision after pars plana vitrectomy with laser coagulation and gas tamponade over a follow-up of 26 months. The purpose of our case report is to emphasize the role of the vitreoretinal interface in the pathogenesis of macular detachment associated with optic pits. CASE REPORT AND THERAPY: A 32-year-Caucasian woman developed macular detachment associated with an optic pit on her right eye. The vision deteriorated to 24/60. A standard three-port pars plana vitrectomy was performed. After creating a posterior vitreous detachment, all vitreoretinal adhesions were removed carefully. Peripapillary laser coagulation and gas tamponade with 15% C2F6-air mixture followed. For 26 months after surgery the macula has been flat. The vision is 60/60. CONCLUSIONS: Besides the exudative component of macular detachment, the vitreoretinal interface seems to play an underestimated role in the pathogenesis of maculopathy associated with optic pits. Tractional forces could explain the delay of macular detachment in young adulthood and the frequency of treatment failure after laser coagulation and gas tamponade. Pars plana vitrectomy with complete removal of all vitreoretinal adhesions should be a suitable technique in the treatment of macular detachment associated with optic pits. PMID- 10827465 TI - [Invasive aspergillosis of the orbit in immunocompetence]. AB - BACKGROUND: An isolated aspergillosis of the sphenoidal sinus is a difficult diagnosis because the often misleading clinical symptoms of this rare disease can develop late and first be presented to the ophthalmologist. Because of this even life-threatening disease in normally immunodeficient patients, fast diagnostics and treatment is indicated. CASE REPORT: We report about an immunocompetent man to suffer from an invasive aspergillosis involving the sphenoidal sinus after external operation of a mucocele. Aspergillus infiltrated the right orbit by bone erosion and led to an acute loss of vision on his right eye and a pathological visual field (visual acuity 0.1). A systemical antimycotic therapy was performed and the right orbit and the optical channel were operated to remove the mycosis. During operation the optical nerve was found to be completely surrounded by the mycosis. After surgery visual acuity recovered slowly but completely in about two weeks (visual acuity 1.0) and visual field examination revealed normal findings. These findings were stable now for one year. CONCLUSION: The main findings of unilateral acute visual loss and pre-operations of the paranasal sinuses in combination with persistent headache can lead to the suspect of an aspergillosis infection. It is important to establish correct diagnosis--especially in immunocompetent patients--early to prevent a probably irreversible visual loss and in an advanced stage to handle the life-threatening risk of this disease. PMID- 10827466 TI - [Ophthalmology in the European Union. Status at the beginning of the 21st century and opening to Eastern Europe]. PMID- 10827467 TI - [Striking scleral pigmentation]. PMID- 10827468 TI - [On line ophthalmologic education. Live transfers and on demand lectures via Internet]. AB - Continuing medical education for ophthalmologist is important and demanded from the board of ophthalmic organizations. The use of new multimedia possibilities provided by the internet, offers an alternative to acquire the training sessions requested by the board. Aim of the paper is to show the efforts made, to place an internet-based training website for ophthalmologists by transferring live ophthalmology events via the internet. The live-transfers of selected lectures is realized by transferring the original sound compatible to the software "Real Audio-Player" with an rate of 8000 Hz (encoding-depth 14 bits). By using this software it is possible to follow the lectures live. It includes also 2 live pictures of 2 digital camcorders to show the appropriate slides from the lecturer. The live broadcasted lectures were afterwards revised and are available at any time as "on-demand lectures" with original sound and slides. Since November 1997, 15 continuing medical education events for ophthalmologists of the University Erlangen-Nurnberg, Germany and from other universities were live broadcasted covering all aspects of phthalmology can be viewed as "on-demand lectures" with original-sound and slides under the internet-addresse http:?www.onjoph.com/deutsch/live.html. The broadcasted events of workshops were watched via the internet with the appropriate software by 3500 persons, and approximately 5200 persons were listening at least one of the lectures since 1997. PMID- 10827469 TI - [Problems of tumor progression: doubts or hopes at the turn of the Millennium?]. AB - After major developments in the diagnosis and therapy of the primary cancer, at the turn of the century clinical oncology is still facing the major challenge: management of the disseminated disease. Cancer biology provided basic information on the genetic and biochemical background of the process, however, it turned out that the individual tumor types use a wide range of mechanisms for invasion and metastasis. Recent major discoveries concerning invasion are identification of the invasion organelle (invadopodia) and identification of certain molecular mechanisms leading to organ-selective metastatization. Prognostic pathology emerged as a new diagnostic field, specialized in predicting the metastatic behavior of the tumor based on the geno- and phenotype of the primary tumor. Ultimately, the first drugs which were designed on the principles of tumor progression entered clinical trials (angiogenesis- and MMP-inhibitors) indicating a slow but steady transfer of cancer biology knowledge to clinical oncology predicting a significant development in the management of disseminated cancer patients. PMID- 10827470 TI - [The cumulative risk index in stroke prevention]. AB - The screening and special management of high risk patient, is possible strategy to reduce the high morbidity and mortality of stroke. In the II. district of Budapest we examined high vascular risk patients cooperating with general practitioners. We let the severity of risk with arithmetic of the cumulative risk index. Significant relation was found between of high cumulative risk index and 50% or higher carotis stenosis, as well as the cumulative risk index and low Se HDL level. Increased blood pressure level has been found in among the treated hypertonic patient. Reduction of high blood pressure was documented in the 6 month control examination. Finally there was very low level of smoking in the high vascular risk patient group. PMID- 10827471 TI - [Alkaline phosphatase level and bone density in Scheuermann's disease and in adolescent idiopathic scoliosis]. AB - Authors made a comparison between 167 suffers from Scheuermann's disease (SD), 70 adolescence idiopathic scoliosis (AIS) and 132 age, sex, height, weight, pubertal developmental stages (Tanner stages) matched controls. The height percentile in 130 cases was also determined. The bone mineral density (BMD) was measured by pQCT on the non dominate sided radial bone. The alkaline phosphatase (AP) level increased at the beginning of puberty and in the puberty in SD. In SD the trabecular Z-score of BMD was significantly decreased in Tanner stage 1 to 4 in both boys and girls. It was not found any significance difference, however, in Tanner stage 5, while in AIS girl no significance decrease of BMD was found. In SD good correlation could be demonstrated between increase in AP and decrease in trabecular Z score r = 0.2, while did not correlate with height percentile. The AP level's increase, and radiomorphometric data of SD severity (intervertebral space narrowing and antero-posterior diameter increase of vertebral body) shows a significant correlation with decreased trabecular Z-score in the period of prepuberty. PMID- 10827472 TI - [Comparison of various methods in the detection of Helicobacter pylori infection]. AB - There are several possible methods to detect H. pylori in the gastric mucosa. The aim of our prospective study was to evaluate the diagnostic value of these tests and to define their place in the clinical practice. 109 (45 male, 64 female) patients who underwent upper GI endoscopy were included. Before endoscopy, whole blood was collected for serological test and 13C-UBT (13C-urea breath test) was performed. During endoscopy, multiple biopsies were collected from the antrum and corpus for the examination of H. pylori status by histology and rapid urease test. Patients with positive histology and a positive result of any other examinations, or--in case of negative histology--with two positive results of the remaining examinations were considered to be H. pylori-positive. 50 patients (46%) proved to be H. pylori-positive. Sensitivity and specificity, respectively, were as follows [in brackets: negative predictive value (NPV) and positive predictive value (PPV), respectively]: histology, 98% and 92% (98% and 91%); 13C UBT, 93% and 98% (94% and 98%); rapid urease test, 83% and 100% (86% and 100%); serological examination, 86% and 74% (88% and 70%). The sensitivity and the clinical role of the methods used for the detection of H, pylori infection is different. Histology is the most reliable method if endoscopy in performed. The positive result of the rapid urease test is also of good diagnostic value. The 13C-UBT is the method of choice if no endoscopy is performed and the clarification of H. pylori status is necessary. This method can be useful to control the success of bacterium eradication as well. The serological examination provides instant result, therefore this method is proposed for screening and epidemiological studies. PMID- 10827473 TI - [Role of bile acids and endotoxins in the pathogenesis and therapy of psoriasis]. AB - The authors demonstrated significant curative effect of bile acids (Suprachol; Acidum dehydrocholicum) in 551 psoriatic patients. The clinical efficiancy was evaluated by means of PASI-score (Psoriasis Area Severity Index). During this treatment (1-8 weeks) 434 patients (78.8 per cent) became asymptomatic. However, the traditional therapy resulted in 62 patients (24.9 per cent) of 249 sick persons a recovery (p < 0.05). In acute form of psoriasis (184 patients) this curative effect of bile acids was elevated (95.1 per cent). Two years later 319 patients (57.9 per cent) of bile treated 551 people were asymptomatic in contrast with 15 people (6.0 per cent) of 249 traditional treated patients (p < 0.05). In same time among the patients which were treated in acute form of psoriasis 10 (7.2 per cent) of 139 controls and 147 (79.9 per cent) of 184 bile-treated individuals were asymptomatic (p < 0.01). On the basis of their clinical observations (digestive disorders, ultrasonical confirmed gallbladder complaints, etc.) authors supposed that the deficiency of bile acids and the consecutive endotoxin translocation might play a role in the pathogenesis of psoriasis. In normal conditions the bile acids as detergent (physico-chemical defense) can protect the body against enteral endotoxins while split them to atoxic fragments and so preventing consecutive cytokin liberation. PMID- 10827474 TI - [In memoriam Endre Kelemen (1921-2000)]. PMID- 10827475 TI - [Ambulatory treatment of hyperthyroidism with radioiodine]. PMID- 10827476 TI - [Ambulatory treatment of hyperthyroidism with radioiodine]. PMID- 10827477 TI - [The century of stress]. PMID- 10827478 TI - [The Nobel Prize in physiology and medicine]. PMID- 10827479 TI - [Physicians without boundaries]. PMID- 10827480 TI - [The ideology of market economy and medicalization]. PMID- 10827481 TI - [On the holy ground of dermatology]. PMID- 10827482 TI - [Influenza, prevention and solidarity]. PMID- 10827483 TI - [Social class and birth weight]. AB - The aim of this study was to investigate whether the birthweight of Scandinavian children varies according to the social class of their parents, especially the mother. Data were taken from the Scandinavian part of an international multicentre study of fetal growth and perinatal outcome. The occupations of the pregnant woman, her spouse and her parents were registered according to the Nordic classification of occupations. This classification has been criticised for being too detailed to be suitable in epidemiological studies, and the data were recorded into the British system of five classes. The birthweight of female newborns in social class V was 301 g lower than in the other social classes (p < 0.05). A corresponding difference was not shown among male newborns. Newborns of women that had migrated downwards in the socioeconomic system, were 117 g lower than if the migration was upwards (p < 0.05). This difference among female newborns was 164 g (p < 0.05). This study demonstrated that there are differences in birthweight according to social class. This may partly be due to genetic factors and a higher prevalence of smoking and high body mass index, i.e. a less favourable lifestyle in the lower social classes. PMID- 10827484 TI - [Phlegmasia cerulea dolens as the initial symptom of abdominal aortic aneurysm]. AB - Phlegmasia cerulea dolens is a serious condition and a result of central venous obstruction which can lead to venous gangrena and amputation of the affected limb. Multiple underlying conditions are described: malignant disease, trauma and hypercoagulable states such as heparin-induced thrombocytopenia, deficiency of protein C and antithrombin III. Phlegmasia cerulea dolens is also described as complication to insertion of a vena caval filter. We present a patient who developed phlegmasia cerulea dolens in his left lower limb as a result of an undiagnosed lumbal aortic aneurysm. To our knowledge this is the first report of phlegmasia cerulea dolens as the initial symptom of an abdominal aneurysm. PMID- 10827485 TI - [Recently discovered herpes viruses explain the etiology of well-known diseases]. AB - Three new members of the family of human herpesviruses (HHVs) have been identified in less than a decade, HHV 6 7 and 8. HHV-6 and HHV-7, both infecting T-lymphocytes and phylogenetically related to cytomegalovirus, were identified as causative agents of exanthema subitum. In addition, HHV-6 has been reported to manifest central nervous system tropism and to be frequently detected in normal brain tissue, but has also been associated with febrile seizures. HHV-7 has been suggested to be involved in the development of pityriasis rosea, but has also been found to occur in normal dermal tissue. HHV-8, related to Epstein-Barr virus and infecting B-lymphocytes, was the first herpesvirus to be identified with molecular techniques. Recent research has been focused on the involvement of proteins expressed by HHV-8 in the pathogenesis of two rare tumours, Kaposi's sarcoma and body-cavity B-cell lymphomas. PMID- 10827486 TI - [Two individuals under the same skin--immune mechanisms in pregnancy]. AB - The fetus may be regarded as an allograft in the maternal organism. This paper gives an overview of pregnancy-associated immune mechanisms, based on the literature and studies performed by the authors. During implantation, maternal tissues are invaded by fetal trophoblasts expressing HLA-G, a trophoblast specific variant of HLA Class I antigens. Recognition of HLA-G stimulates uterine natural killer cells to cytokine production, by which an intrauterine immunosuppression is established. Development, growth and differentiation of placenta is regulated by the cytokines produced. Uterine leukocyte population and expression of cytokine receptors in placental tissues varies throughout gestation, and the complex interplay between trophoblasts and uterine cells, involving a number of cytokines, cytokine receptors, adhesion molecules, enzymes and hormones, changes with gestation. Some cytokines, such as tumour necrosis factor and interleukin-1, may threathen the reproductive process and fetal well being in high doses. A tight regulation of cytokine activities is probably obtained by the observed upregulation of endogenous cytokine buffer mechanisms in pregnancy. The reproductive success and phenomenons like implantation, placental growth and development, maintenance of pregnancy and delivery, appear to rely on complex, gestational age related interplay between cells of fetal origin and the maternal immune system. PMID- 10827487 TI - [Analytical uncertainty--how wrong can a laboratory result be?]. AB - Some uncertainty encumbers the outcome of all laboratory tests. Patient conditions and handling of specimens, as well as analytical variation and systematic error will affect the results. In order not to confuse the interpretation of test results, the analytical standard deviation should not exceed one half of the intraindividual biological standard deviation, and systematic error should not exceed 1/16 of the reference interval. However, these goals cannot always be achieved. Moreover, analytical control procedures have limited ability to detect errors in the analytical process. After one analysis of control material, using +/- 2 standard deviations (analytical) from the mean as acceptance limits, the magnitude of a systematic error must be 3.3 times the analytical standard deviation in order to be detected with 90% probability. As a result of such error, patients' results can be released with a total error up to five times the analytical standard deviation. More complicated control procedures may give smaller total error. Practising physicians should be familiar with the variations in lab results, and interpret the results accordingly. Lab results that conflict with results of other investigations should be used with caution. PMID- 10827488 TI - [Health status in Norway around the year 1000]. AB - One thousand years is a very long period, especially in relation to medical history. More than 30 generations have lived over this space of time, and their thoughts and attitudes and their views on hygiene, sickness and treatment are heterogeneous. This paper will discuss the state of health in Norway at the time of the turn of the first millennium--at a time when Norway underwent profound political, social and religious changes, as well as structure of diseases. PMID- 10827489 TI - [Health, medicine and population development in Norway]. AB - If health, medicine and medical services in Norway throughout the millennium are seen through a demographer's eyes, it is obvious that the health situation and the preconditions for medical work have changed profoundly, especially in the last two centuries. A relatively stable, young, and geographically dispersed population, living with, and probably to a large extent accepting high morbidity and high mortality, has gradually been transformed into a more middleaged, urbanized population where absence of disease and untimely death is perceived as a normal situation, which it is the goal of the health services to achieve. The population growth was especially accelerated during the 19th century, when mortality rates declined rapidly before a corresponding decrease took place in the birth rates. The demographic effects of medicine in Norway in the 19th century should mostly be ascribed to efforts in preventive and social medicine. The life saving and life prolonging effects of curative medicine belong to the the 20th century, a period when specific treatment of infections had become available. However, further achievements in demographic effects of medicine have to take into account the demands set by the perceptions of health and welfare in the population, and the ever increasing social and economic constraints. PMID- 10827490 TI - [What shapes people's notions of health and illness?]. AB - Lay conceptions of health are shaped by a variety of demographic, social, and personal factors. Research has focused on psychological and sociological perspectives, and three core models have dominated the field: the cognitive developmental model, the health locus of control model, and the so-called health belief model. Societal factors have been less studied. On the basis of a meta model we discuss how changes in prevalence of disease and disease panorama, social network, the experience of uncertainty, and the medical focus on risk may have impact on lay conceptions of health and disease. The joint effect of the medical focus on risk and disintegrating social networks seems to be the central force behind medicalization. PMID- 10827491 TI - [The epidemiological transition during the 20th century]. AB - An epidemiological transition took place during the 20th century causing profound demographic changes in the industrialised world. The transition is a reflection of lower birth rates and changing disease rates. These changes are associated with improved nutrition, hygiene and other social factors, and are unevenly distributed according to socio-economic status. Preventive efforts, intentional or not, and diagnostic and therapeutic improvements will contribute to a demographic transition resulting in an increasing number of elderly people. The epidemiological transition and the development of more sophisticated medical techniques will increase the demand for health care. PMID- 10827492 TI - [Health services--a system with a variety of objectives]. AB - The article presents the objectives of the Norwegian health care system as they have been outlined in governmental publications from 1672 to 1999. A model for analysis of objectives is presented. Concern for the patient, estimated medical needs in society and administrative guidelines are common elements in the set of objectives through the period. Effective use of resources became a concern at the end of the period. The stated objectives of the health care systems of Norway are more concerned with the health care system than with health in itself. The model for analysis uses the concepts of mercy, benefit, order and profit for providers to classify the various objectives. Evaluated by this model, the main objectives seem to have been surprisingly stable throughout the period covered by the study, though the concept of benefit seems to become a more important consideration during later decades. PMID- 10827493 TI - [Growth and stagnation--development of health services during the 20th century]. AB - The health services of Norway have changed over the last century in tune with the development of society. The health services are not only the result of scientific and technological progress, but also a product of changes in prevailing political and social conditions and ideologies. Quality assessment and concern for the ethical basis of the health services ought to be given priority in the years to come. PMID- 10827494 TI - [Welfare and health in the century of social security]. AB - Great achievements for public health in this century include penicillin, oral contraception, vaccination and transplantation, but the greatest contribution to the health of Norwegians may have been made by social security and the welfare state policies. The beneficial effects of social security include: less social inequality, a factor which in itself makes for better health, some degree of financial security for people who are ill and unable to support themselves or their families, universal availability of medical and health services. This paper explores the ideas and sources from which present-day social security legislation stems. The emergence of state based welfare legislation was inspired by developments in Germany under Bismarck; accident insurance for industrial workers was the first to be introduced in 1894. Several periods of active social security legislation followed at times when labour unions and socialist ideas were strong, supported, to some degree, by radical clergy and public health doctors. Social democratic governments, inspired by the British Beveridge plan towards the end of World War II, took the lead during the long post-war period until the mid 1970s. The Scandinavian model of social security is based on universal coverage and a single payer system. However, since the 1980s, as in other countries of Northern Europe, the social security system has come under pressure from market liberalism. Now at the turn of the century, increasing social inequalities cause some concern for the future of the welfare state. PMID- 10827495 TI - [Defending pluralism]. AB - The 20th century is the century of modernity. Medical science and technology have basically and positively changed the health of the population. The serious infectious diseases are under control and the health services and their institutions have expanded extensively, in Norway as throughout the western world. Nevertheless, there is an impression of general frustration and discontentment. The authors reflect over this paradox by reminding the reader of some of the main developments in primary care in this century. They call attention to former Director General of the Norwegian Board of Health, Karl Evang, and his role as initiator and leader, and see him in contrast to former Minister of Health, Werner Christie and his criticism of the system. The authors suggest that the crisis within the health care system emerging over the last decades, should be understood as a crisis of modernity. There is a need to adjust some of the ideals which have dominated our century. In order to face the challenges of post-modern society, they recommend restoring the art of medicine. Above all, there is a need to incorporate pluralism as an essential value in the planning of our future health care system. PMID- 10827496 TI - [The history of the asylum--the history of "the great confinement"?]. AB - Histoire de la Folie (1961) by the French philosopher Michel Foucault has profoundly influenced the general public's conception of the history of psychiatry, but the book is relatively seldom discussed among psychiatrists. Foucault views the core characteristics of psychiatry in the light of what he terms "the great confinement" of the mad and other outsiders. He maintains that the confinement took place all over central Europe from 1650 until 1800, and that the psychiatric asylum was designed on the basis of the confinement house. This paper discusses whether this great confinement actually took place and critically examines Foucault's interpretational model. There is a further discussion of the main reasons why the humanistic and creative spirit characteristic of the early nineteenth century asylum was replaced by stagnation and disillusionment 150 years later. PMID- 10827497 TI - [Historical retrospect on women's health in our century]. AB - At this turn of the century, we may look back on a hundred years of major social changes with a huge impact on women's health, many of them related to women's role in reproduction. Effective contraception has become available to all women, and access to safe abortion has been secured through legislation. The majority of women are now in paid employment and women's position in society is dramatically improved. The decrease in mortality has affected women and men differently. Women live longer than men, but they tend to be afflicted by chronic non-lethal diseases. Issues which earlier were kept out of the public debate on women's health are now part of that debate. The improvements in women's health are not only a consequence of changing living conditions and better health services; they are also linked to changing attitudes to sexuality, love and marriage. PMID- 10827498 TI - [Health as premise for urban planning]. AB - Concerns for health have been a foundation for planning in towns the cities in Norway from the 19th century up to the present. Doctors previously took active part in discussions of urban planning. Planners developed new concepts for urban design and town planning. Sun, air, low density and separation of functions were important tools to achieve a healthy city. Modern planning has led to dispersed cities requiring a great deal of transport. A more dense built-up urban area is necessary to achieve sustainable development. Higher population density and a mix of functions should be encouraged in order to reduce the need for transport. This strategy is contrary to the development of urban planning over the last 150 years. Health professionals must participate in the discussion on urban form and planning in order to avoid new health problems higher density and a mix of functions in our cities may cause. PMID- 10827499 TI - [Work and health--challenges at the turn of the century]. AB - This article gives an overview of a century of developments in the field of work related health; relevant legislation, the incidence of work-related accidents and diseases, and the development of occupational health services. It is difficult to evaluate the working environment in Norway, as official statistics concerning work-related accidents and diseases are incomplete. Such data are of major importance for assessments of the effect of interventions in the working environment; hence, improved statistics should be a high priority in the future. There is a need for a national plan for work and health and for devoting more resources to the Labour Inspectorate. The occupational health service is a very good arena for preventive health work; it ought to be better integrated in the Norwegian general preventive health efforts. PMID- 10827500 TI - [The outer environment--now and then]. AB - Hazardous environmental factors in Norway have changed considerably over the last decades, often for the better. During the last five-year period, water-works serving 800,000 Norwegians have been renovated. The earlier high levels of sulphur dioxide and lead in urban air are now very low, whereas suspended particulate matter and nitrogen oxides continue to be at levels which can induce adverse health effects. Radon and tobacco smoke are now important indoor contaminants. Moisture-induced damage in dwellings may lead to health problems; the extent of such damage is, however, not known. A number of fjords are still contaminated with metals, PAH (polycyclic aromatic hydrocarbons), PCB (polychlorinated biphenyls) and dioxins, even though industrial discharge to water and air has been greatly reduced. Body burdens of DDT, PCB and dioxins have been markedly lowered over time. There is a continuous increase in the use of chemicals, though the chemicals are better tested and controlled than before. The total volume of pesticides used has fallen over the last three decades. There are no clear changes over the last 15 to 20 years in perceived noise exposure. PMID- 10827501 TI - [Cobalt mining factory--diagnoses 1822-32]. AB - The cobalt mining company at Modum in Norway had its own health service. The doctors kept records of all patient contacts. Recently discovered material from the 1822-32 period shows that gastrointestinal diseases, injuries and respiratory diseases made up the majority of patient contacts, in addition to unspecified infections. PMID- 10827502 TI - [Health commissions' activities in five rural municipalities in Telemark between 1861-1900]. AB - It has been assumed that local health administrations in rural areas in the second half of the nineteenth century functioned in a rather passive and ineffective way. The present study focuses on the activity of the health commissions in the municipalities of Vinje, Seljord, Hitterdal, Gjerpen and Bamble. In the 1860s og 1870s, few meetings were held. Lack of interest and communication problems between doctors (who chaired the commissions) and the municipal councils were pointed out as causes. In commission meetings, the doctors used most of the time to give information on the present health situation in the area and on general medical issues. After 1885 the activity in the commissions gradually increased. New legislation and health regulations gave the health commissions a more important role in public health administration. In Hitterdal and Seljord the health commissions contributed actively to fighting the diphteria epidemics in the late 1880s. With urbanization and industrialisation in lower Telemark, environmental health issues took on added importance for the health commissions of Gjerpen and Bamble. The problems were much more effectively dealt with in Bamble than in Gjerpen, possible because of differences in the organisation of the commissions and in the workload for the doctors who chaired them. PMID- 10827503 TI - [Health status and living conditions in medical district of Sandnes during the period 1866-1900]. AB - The municipality of Sandnes became a separate medical district in 1866. The district physicians' reports give us an insight into health and living conditions in this district in the last half of the 19th century. This paper is based on a study of all statistical data and physicians' comments concerning the Sandnes district for the 1866-1900 period. The incidence of various diseases is estimated. Mental illness seems to have been more frequent in Sandness than in the rest of the county. In the physicians' reports, this was explained by the dominance of religious Puritanism in the district. The incidence of other disease, such as typhoid fever, tuberculosis and scarlet fever, did not differ essentially between Sandnes and the county as a whole. The living conditions in the district were rather good during the whole period, though the physicians' reports provide interesting insight in changes from one year to another. PMID- 10827504 TI - [Obstetrics--a women's discipline for men]. AB - In recent years there has been a considerable increase in the number of female gynaecologists in Norway. This contrasts sharply with the difficulties pioneering Norwegian female physicians had in entering this specialty. Three generations ago, leading Norwegian physicians argued against female gynaecologists. This article describes the controversy in 1914 when Dr. Louise Isachsen (1875-1932) claimed discrimination when she was not appointed senior registrar at the National Hospital's Midwifery Clinic. Her case was fiercely debated, not only in the medical profession, but also in newspapers and in Parliament. The clinic's director, Professor Kristian Brandt (1859-1932), found women unsuited for operative gynaecology and emergency obstetrics. We compare his arguments against female physicians with the requirements for Norwegian midwives, all female at the time. Brandt himself played a crucial role in formulating these requirements through his teaching and textbooks. PMID- 10827505 TI - [Multiple benign sarcoid of the skin--100 years since Caesar Boeck's pioneering article]. AB - In December 1899, Professor Caesar Boeck (1845-1917) in Kristiania (now Oslo) published his pioneering article on "Multiple benign sarkoid of the skin". At the time, Boeck's sarcoid was a dermatological curiosity, but in his latest work in 1916, Boeck recognised the generalised character of the disease. Today, 100 years later, sarcoidosis is still an enigmatic disease. Caesar Boeck was a hard-working clinician and histologist, the founder of modern dermatovenerology in Norway. This article gives an account of Caesar Boeck's family background, his scientific career and his personality. PMID- 10827506 TI - [Medical humor--theoretical basis and practical use]. PMID- 10827507 TI - [Is the general practice system based on men's premises? The pressure against the clock as a stressful factor in the physician-patient relation]. PMID- 10827508 TI - [Quality of life after organ transplantation]. PMID- 10827509 TI - [The increasing costs of mammography]. PMID- 10827510 TI - [Breast feeding and alcohol drinking]. PMID- 10827511 TI - [Nicotine level measurements in restaurants in Tromso 1999]. PMID- 10827512 TI - [The phenomenon of "earth radiation" and disease]. PMID- 10827513 TI - [Belief and health]. PMID- 10827514 TI - [Just talk?]. PMID- 10827515 TI - [Internship and residency]. PMID- 10827516 TI - [Oral contraceptives and thrombosis--an adverse effect of mandatory notification?]. PMID- 10827517 TI - [School injuries among 7 to 15 years old children]. AB - This study had two purposes: first, to describe the incidence and characteristics of school injuries among 7 to 15 year old pupils in Norway; second, to identify effective preventive measures. Data on school injuries were taken from the Norwegian National Injury Register (National Institute of Public Health) for the 1995-97 period, n = 2,975. The incidence of school injuries was 29 per 1,000 children-years. Over the nine-year age span, boys had a higher cumulative incidence (292 injuries per 1,000 boys) than girls (239 injuries per 1,000 girls); 1% of the injuries was categorized as severe. Among pupils aged 7-12, 34% of injuries occurred during sports activities; among the 13-15 age group, 60% were related to sports. A literature review was conducted in order to identify preventive measures shown to be effective in reducing school injuries. The databases used were Cochrane Library (issue 4, 1998), Medline (1966-98) and Embase (1980-98). However, no studies found in the literature had evaluated preventive measures. On the basis of our findings, we suggest that preventive measures should emphasise non-sport injuries among the younger pupils and sport injuries among the older pupils. Preventive measures should be evaluated. PMID- 10827518 TI - [Invasive pneumococcal infections in children from Troms, Nordland and Sor Trondelag 1980-95]. AB - Invasive pneumococcal infections may be severe. We have examined epidemiology, risk factors and outcome of these infections. In the years 1980-95, 76 children below the age of 15 with invasive pneumococcal infections were admitted to the hospitals in the counties of Troms, Nordland and Sor-Trondelag in Norway. The incidence rate in children 0-2 years old was 10.3 cases per 100,000 persons per year, and 1.8 in children 3-14 years. Of the patients 24 had meningitis and 52 bacteraemia. All bacteriological isolates were sensitive to benzylpenicillin. Seven patients died and five developed sequelae. Thirty-one of the children had risk factors prior to the infection. Children with hypo- or hyperventilation at the time of arrival, and/or impaired circulation fared worse than those with normal findings. Children with underlying risk factors have a much higher frequency of invasive pneumococcal infections than other children. Patients who had impaired circulation or ventilation on admission have a bad prognosis for healthy survival. PMID- 10827519 TI - [Fulminant influenza pneumonia]. AB - Severe community acquired pneumonia is a common cause of acute respiratory failure requiring ventilator support. Bacteria are by far the most common pathogens, necessitating initial coverage with antibacterial agents. When influenza virus is involved in the aetiology of severe pneumonia, its role in the pathogenesis is most often that of facilitating secondary bacterial pneumonia. The influenza virus itself can, however, cause a severe and rapidly evolving viral pneumonia. We describe two cases of fulminant pneumonia with multiorgan failure, in which extensive microbiologic and serologic diagnostic test did not reveal other causes for the pneumonia than influenza virus. Different clinical lung manifestations of influenza are discussed. Influenza must be considered as an aetiologic possibility in acute respiratory failure. PMID- 10827520 TI - [Prediction of developmental problems and functional disabilities in infancy]. AB - Methods to identify developmental problems in children have been requested for a long time. We wanted to find out whether high scores of cumulative non-optimality during infancy indicated disability or developmental problems. In a prospective study, a population of children (n = 1,217) was followed from pregnancy until the age of four. Medical, motor, and neurological data, as well as information about attention and social interaction were added in a cumulative non-optimality score. Information about family and environment was also included. When the children were four years of age, multiple methods were used to identify those with disability and developmental problems. Complete data were obtained for 926 children. The predictive values of the instrument were based on children diagnosed as functionally disabled at the age of four (n = 32). The sensitivity was 87.5%, specificity 93%, positive predictive value 30.8% and negative predictive value was 99.5%. These values are higher than those of comparable methods. The method could be implemented in the normal Norwegian "child health centre" without difficulty. The discussion in this article focuses on whether or not the method should be implemented in the ordinary health care system. PMID- 10827521 TI - [Hypokalemic pareses secondary to renal tubular acidosis]. AB - A 24 year old woman presented with flaccid paralysis, severe hypokalaemia and hyperchloremia, metabolic acidosis. Immunological tests and labial glandular biopsy indicated primary Sjogren's syndrome as the underlying cause of her distal renal tubular acidosis. The patient recovered after alkali and potassium substitution and was put on oral treatment with potassium citrate. PMID- 10827522 TI - [Pulmonary embolism among young non-pregnant women]. AB - This study focuses on the incidence and risk profile among young women with a diagnosis of pulmonary embolism over the 10-year period 1988-97. A total of 66 non-pregnant women aged 15-44, treated for the first event of pulmonary embolism, were identified by discharge diagnosis at two main Norwegian hospitals (The Regional Hospital of Trondheim and Ulleval Hospital, Oslo). The estimated overall incidence of pulmonary embolism was 6.8 per 100,000 women-years. The crude incidence decreased from 7.5 per 100,000 women-years in 1988-90 to 4.1 per 100,000 women-years in 1996-97 (p < 0.1). 25 (38%) of the 66 patients were identified with at least one medical risk factor, four patients (6%) with a family history of thrombophilia and 30 patients (45%) used oral contraceptives (OC) at the time of diagnosis. 25 (68%) of the 37 patients without registered risk factors used OC. The incidence of pulmonary embolism decreased during the 10 year period, with unchanged risk profile. Two women (3%) died from pulmonary emboli shorter than three weeks after onset of symptoms. PMID- 10827523 TI - [Deep venous thrombosis in young women in Norway]. AB - The incidence and distribution of risk factors in non-pregnant women diagnosed with deep vein thrombosis is not well known and was the target of the present study. The medical records of 102 non-pregnant women aged 15-44 hospitalized with deep vein thrombosis at two major Norwegian hospitals were examined retrospectively. The overall incidence of first event of deep vein thrombosis remained fairly constant over the 10-year study period and was estimated at 10.5 per 100,000 women-years. The incidence increased with age and was found twice as high for women 35-44 years of age when compared with the 15-34 age group. Forty three (42%) of the patients had no identifiable risk factors. Surgery or trauma was found in 37 (36%) of the patients, eight (8%) had a diagnosis of associated cancer, and five (5%) were drug addicts. Five of eight patients registered with immobilization had immobilizing chronic neurological diseases. Thirty-nine (38%) of the patients used oral contraceptives at the time of diagnosis, and 14 of the 39 users of oral contraceptives were identified with medical risk factors. Over the 10-year study period there was no change in the incidence of venous thromboembolism and the risk profile among cases remained constant. PMID- 10827524 TI - [Reporting of adverse drug reactions to the Norwegian Drug Control Agency]. AB - This article reports on a retrospective study in two major Norwegian hospitals of spontaneous reporting to the Adverse Drug Reaction Committee of the Norwegian Medicines Control Agency of adverse reactions from the use of oral contraceptives (OCs) among patients with a diagnosis of venous thromboembolism. In these two hospitals, 168 women between 15 and 44 were diagnosed with a first event of venous thromboembolism during the ten-year period up to the end og 1997; 69 of them were OC users at the time of diagnosis. Three medical records (4%) kept at the hospital contained information on a report of adverse drug reaction to the Norwegian Medicines Control Agency. The Agency's database of all spontaneous reports on adverse drug reactions contains 112 reports of venous thromboembolism in the context of OC use. These reports came from all over Norway during the ten year period of our study; four of the reports came from the study hospitals. We conclude that venous thromboembolism during the use of OCs was underreported by more than 90%. This corresponds to estimates of underreporting of other adverse drug reactions from the use of various compounds, in Norway as well as in other countries. PMID- 10827525 TI - [Sale of oral contraceptives, births and abortions prior to and after the "Marvelon issue"]. AB - This study focuses on the situation in Norway after the UK Committee on Safety of Medicines issued an alert on an increased risk of venous thromboembolism with third-generation OCs on 18 October 1995. The Norwegian Medicine Control. Agency immediately issued a press release warning Norwegian women about the health risk associated with third-generation OCs (in Norway, the drug Marvelon only). In this study the effects of the Norwegian warning are evaluated by the use of data on the sale of OCs, number of induced abortions, and births by quarter for the 1992 to 1998 period. From third quarter 1995 to first quarter 1996, sales of Marvelon to pharmacies dropped by 75%, while total sales of OCs were reduced by 5%. From late October 1995 thorough January 1996, approximately 50,000 women stopped taking Marvelon. A maximum increase of 200 induced abortions could be attributed to the negative news on OCs in October and November 1995. There was no subsequent change in the birth pattern. These data confirm that Norwegian women who shifted to another type or stopped taking OCs during the 1995 media crisis avoided unplanned pregnancies. PMID- 10827526 TI - [Cancer and religiosity--a follow up of patients with Hodgkin's disease]. AB - In this study of religiosity among cancer patients and the relationship between religiosity and illness, a questionnaire was sent to 183 survivors of Hodgkin's disease in November 1995, all of them patients treated at the Norwegian Radium Hospital between 1988 and 1994. 107 patients (58%) gave replies to 45 questions relating to religion and belief, view of life, quality of life, and the relationship between religiosity and illness. The patients' attitude to religion differed little from that of the Norwegian population at large. 15% of the patients defined themselves as atheists, 14% as agnostics, 23% believed in God as an impersonal supreme power, and 48% in a personal God. 40 patients (38%) had changed their religious belief, 33 of them becoming more religious. 58% had prayed to God (or a supreme power) for cure. Half of the respondents felt that their religious belief had been a support during their illness. 52 (49%) were not satisfied with the pastoral services during hospitalisation. The study indicates that cancer disease activates religiosity, and that religiosity may help patients cope with their disease. PMID- 10827527 TI - [What is the most important for the patient in the meeting with a general practitioner?]. AB - Efficient medical communication depends on an understanding of the patient perspective. The purpose of this study was to characterise the most important consultation elements as perceived by primary care patients. The experience of 31 patients regarding their most recent medical encounter were recorded in six focus groups. All groups were audiotaped and analysed by qualitative methodology. Five consultation themes emerged spontaneously in all groups: Interaction between doctor and patient (e.g. communication, emotional experiences, and personal relationship with the doctor) and consultation outcome were the most important elements. Information, continuity of care, and time with the doctor were considered reasonably important. Six other themes received limited attention in a few groups only (e.g. availability, doctor's gender, time in the waiting room, auxiliary personnel, the clinical examination, and technical skills). Patients value the personal, attentive, and listening doctor. The feeling of not having to hurry during the consultation is more important than the actual number of minutes. Patients' level of emotional involvement and their specific expectations are often undisclosed. There are large individual differences with regard to the meaning of and need for information. PMID- 10827528 TI - [Words and language used by patients when describing consultation with general practitioners]. AB - Increased knowledge of how patients express their experiences may help physicians to capture their patients' perceptions and agenda. The aim of the present study was to examine words and language used by patients when describing specific primary care consultation experiences. Six focus group sessions with a total of 31 patients were conducted shortly after a general practice consultation. All group sessions were audiotaped and analysed by qualitative ethnographic methodology. Seven distinct grammatical and verbal characteristics were found: The consultations were recited in the first person with little recollection of the doctors' talk or actions. Subjectivity was constantly underlined by the verbs "feel" or "think". When expressing negative emotions, the personal pronoun constantly was changed from the personal singular to the impersonal or plural form. Language tended to be evasive in connection with possible criticism. Negative emotions were diminished by the use of limiting expressions and modest wording. Non-committal expressions, with a wide range of possible meanings, were frequent, and were possible to decipher only by close attention to non-verbal cues. We conclude that doctors may capture more of their patients' hidden emotional messages in the consultation by increased awareness of specific verbal characteristics and non-verbal cues. PMID- 10827529 TI - [Morbidity and mortality in pandemic influenza in Norway]. AB - This article is partly based on a newly published influenza pandemic preparedness plan and risk analysis for future pandemics in Norway, and presents estimates on morbidity and mortality in influenza pandemics in Norway in the 20th century. In addition, estimates on morbidity and mortality for a future pandemic are given. One of the main conclusions is that excess mortality in connection with Spanish influenza and Asian influenza is highly underestimated. The proportion of the infected that died in the pandemics was low, and was highest for Spanish influenza. The morbidity will be highest if a pandemic similar to Asian influenza reappears, while mortality will be highest if a Spanish flu is repeated. In the worst case scenario for a pandemic today, based on the figures from the Spanish flu, the number of excess deaths could reach 29,000, an increase of 60% compared to the average annual number of deaths seen in the 1990s. PMID- 10827530 TI - [Pseudo-Meigs' syndrome]. AB - Cytologic examination of the body cavity effusions in patients with ovarian tumours is performed to differentiate between reactive processes and tumour spread. While detection of malignant cells is a marker of metastatic disease and a sign of bad prognosis, benign effusions affect neither disease stage nor the patient's prognosis. Determination of the presence or absence of tumour spread is based primarily on cellular morphology. As distinction between reactive mesothelial and cancer cells can be difficult, immunocytochemistry may be employed in equivocal cases. The case of a 42-year-old woman who presented with a large pelvic mass accompanied by ascites and hydrothorax is described. Cytomorphology of preoperative pleural fluid specimen was inconclusive. Immunocytochemical examination of cell block sections using: BerEP4, B72.3, CA 125, CD15, CEA, E-cadherin and calretinin was done. No epithelial cells were detected and diagnosis of reactive mesothelial cells was made. Laparotomy was performed and adnexal tumour removed. Borderline mucinous tumour of the ovary was diagnosed. There was no recurrence of the ascites or hydrothorax. The clinicopathologic features and terminology of pseudo-Meigs' syndrome are briefly reviewed. The role of ancillary studies in diagnosis of body cavity effusions is emphasized. PMID- 10827531 TI - [Serous effusions]. AB - Malignant cells in serous effusions are most often found in patients with primary ovarian or breast carcinomas; the prognosis is generally poor. Identifying malignant cells in effusions may be difficult, as mesothelial cells often mimic carcinoma cells. The present data are based on retrieval from the medical databases Medline and Cancer-line. Several ancillary methods can aid in the morphological diagnosis of effusion specimens. These consist of histochemistry, immunocytochemistry, flow cytometry, electron microscopy and molecular biology techniques. Immunocytochemistry is the most widely used technique to date and utilizes both epithelial and mesothelial markers. Electron microscopy is considered to be the gold standard in serous effusion diagnostics, but is of limited use as it is unavailable in many laboratories. The use of molecular techniques may offer an additional possibility of enhancing the diagnostic accuracy and may provide information regarding the origin of the primary tumour. Accurate diagnosis of serous effusion mandates the use of ancillary methods such as immunocytochemistry and molecular techniques, and may aid the clinician in patient management. PMID- 10827532 TI - [To be emotionally affected during medical education]. PMID- 10827533 TI - [Influenza--preventive measures without effect?]. PMID- 10827534 TI - [DNA tests and physician's role]. PMID- 10827535 TI - [Meniere's disease--therapeutic aspects]. PMID- 10827537 TI - [We should reduce adverse incidents and increase safety in health care. The topic of the BMJ from March 18, 2000: Reducing error and improving safety]. PMID- 10827536 TI - [Adverse incidents in hospitals. A review of three foreign studies]. PMID- 10827538 TI - [Hepatic encephalopathy and brain edema]. PMID- 10827539 TI - [Quantitative ultrasound bone measurements]. AB - Quantitative ultrasound (QUS) bone measurement is a relatively new technique for the diagnosis of osteoporosis which is cheaper and easier to use than the more established method of bone densitometry (measurement of bone mineral density, BMD) by x-ray absorptiometry. The two QUS parameters currently measured are BUA (broadband ultrasound attenuation) and SOS (speed of sound). The reported age related changes for healthy women range from -0.27% to -1.62% per year for BUA and from -0.06% to -0.19% per year for SOS. Precision ranges from 1.0-3.8% (CV) for BUA and 0.19-0.30% (CV) for SOS. QUS is believed to reflect mainly BMD. However, studies indicate that QUS also reflects trabecular orientation and other determinants of bone strength independently of BMD. In both cross sectional and prospective studies, QUS seems to be as good a predictor of osteoporotic fractures as BMD. PMID- 10827540 TI - [DiGeorge syndrome. Velocardiofacial syndrome/chromosome 22q11 deletion syndrome]. AB - Patients with a deletion of chromosome band 22q11 are described as having DiGeorges syndrome, velocardiofacial syndrome or chromosome 22q11 deletion syndrome depending on clinical manifestations. The patients have variable severity and combinations of conotruncal heart defects, abnormalities of the ear and palate, facial dysmorphism and mental retardation as well as partial or complete aplasia/hypoplasia of the thymus and endocrine dysfunction, e.g. hypoparathyroidism. The patients may present with impaired immune function, heart failure, hypocalcaemia, facial dysmorphism, impaired hearing and mental retardation. The syndrome, which is a significant cause of heart and craniofacial defects as well as mental retardation, is probably underdiagnosed. In each of the above mentioned phenotypical presentations, chromosome 22q11 deletion syndrome should be considered. PMID- 10827541 TI - [Meniere's disease. The clinical effect of gentamicin treatment]. AB - In patients with Meniere's disease attacks of vertigo can be invalidating. We have tested the effect of instillation of gentamycin in the middle ear to minimize or stop the attacks of vertigo. The results of a prospective investigation of 14 patients with Meniere's disease treated with gentamycin in the period from 1993 to 1996 are presented. The patients were followed up to 53 months after the treatment and a telephone follow-up was performed. At the last follow-up nine out of 12 patients were free of attacks of vertigo. Two improved and one patient was unchanged. In two patients deafness developed in the treated ear, in the other patients the hearing was unchanged measured by speech reception threshold and discrimination loss. It is concluded that treatment of patients with Meniere's disease by instillation of gentamycin into the middle ear does have a therapeutic effect. The treatment is simple and the effect of treatment is equal to other types of treatments. The risk of deafness in the treated ears stresses the importance of giving the patient sufficient information before treatment. PMID- 10827542 TI - [Microbiological examination of patients hospitalized in Aarhus municipal hospital]. AB - The purpose of this retrospective study was to evaluate 1) the nature, extent and relevance of the microbiological examination performed before institution of antibiotic treatment, 2) the relevance of the instituted treatment in the light of the microbiological findings or clinical diagnosis of infection. A minimum of requirements concerning microbiological examination was put forward for each clinical diagnosis of infection. Among patients with the diagnosis of sepsis, cystitis or pneumonia, 33%, 80% and 6.5%, respectively, fulfilled the minimum requirements, no attempt at microbiological examination was performed in 4.8%, 10%, and 32%, respectively. There is a need for education in relevant use of clinical microbiology e.g. samples related to focus of infection, and more consistency in the use of microbiological results. PMID- 10827543 TI - [Aftercare of malleolar fractures in Denmark. A questionnaire study]. AB - A questionnaire was sent out to 59 orthopaedic departments. Seventy-eight percent returned the questionnaire. Four thousand two hundred ankle fractures are treated annually. Fifty percent are operated on (range 12.5-83%). Large departments are on average more surgically active than smaller ones. The noticeable difference, however, in surgical frequency is not connected to size, function or location of orthopaedic department. All fractures are usually immobilized with a circular plaster bandage. Thirty percent use dynamic bandages for some stable ankle fractures. Eighty-five percent bandage for six weeks, 15% for up to eight weeks. Very few allow early mobilization of surgically treated fractures (6%). The rest do not allow mobilization until at least three weeks after surgery. Stable fractures are more frequently permitted immediate mobilization. Several randomized studies recommend that all ankle fractures should be mobilized immediately, surgically treated in a circular plaster bandage, stable fractures in dynamic bandages. PMID- 10827544 TI - [Quality assessment of resuscitation]. AB - The object of the study was to evaluate the availability of paediatricians for resuscitation of newborns and the quality of resuscitation. During the period 1.9.95-15.1.96 a prospective registration was made of the initial treatment of all newborns where a paediatrician was involved. It was noted whether the doctor was present when needed. A comparison with the written guidelines was made. The paediatrician was called to 27% of the 1461 babies born during that period. In 93% of the deliveries with maternal risk factors the pediatrician was called before the delivery and was present at the delivery in 89% of the cases. The resuscitation was optimal in 60%, less satisfactory in 38% and unsatisfactory in 2% of the cases. Intensified training and supervision of doctors dealing with resuscitation of newborns and strengthening of the staff on duty is recommended. PMID- 10827545 TI - [DiGeorge syndrome diagnosed in a 39-year old woman with chronic hypocalcemia]. AB - A 39 year-old woman was diagnosed with DiGeorge's syndrome based on newly diagnosed hypocalcaemia, appearance and history. The patient had congenital cardiovascular malformations, mild mental retardation and ear malformations, and during infancy and childhood suffered from failure to thrive and frequent infections. In children with conotruncal heart malformation, hypocalcaemia and hypoplasia of the thymus in combination it is estimated that chromosome 22q11 deletion is present in almost 100%. The syndrome is probably underdiagnosed. PMID- 10827546 TI - [Eosinophilic enteritis]. AB - We present a case of eosinophilic enteritis in a 45 year-old male with clinical and radiological signs of stenotic inflammatory ileal disease. A diagnosis of Crohn's disease was considered. He developed small bowel obstruction and sixty cm of obstructed ileum was resected. Histopathological examination revealed the diagnosis of eosinophilic enteritis primarily localized to the tunica muscularis. One year postoperatively he relapsed and small bowel X-ray demonstrated 1 m narrow and irregular ileum. He was treated with mesalamine, azathioprine, and cromoglicate, went into remission and fares well one and a half years later. PMID- 10827547 TI - [Picture of the month. Contact dermatitis of the hand]. PMID- 10827548 TI - [Genetic prevention of cancer]. PMID- 10827549 TI - [Adverse events and accreditation by the H:S]. PMID- 10827550 TI - [Hearing tests in specialty practice]. PMID- 10827551 TI - [War or peace between otorhinolaryngologists and hospital audiologists?]. PMID- 10827552 TI - [Hearing tests in specialty practice prior the use of hearing aids]. PMID- 10827553 TI - [Diagnosis of enterobiasis]. PMID- 10827554 TI - [Kirsten Lyloff on death causes of German immigrant children]. PMID- 10827555 TI - [The failure of care]. PMID- 10827557 TI - [What is the best way to assure the quality of patient/physician contact?]. PMID- 10827556 TI - [Kombucha tea]. PMID- 10827558 TI - [The elderly and infections: old friends or sworn enemies?]. PMID- 10827559 TI - [Repetitive transcranial magnetic stimulation. A method in the treatment of depressions]. AB - Transcranial magnetic stimulation (TMS) has been used as a diagnostic tool in neurology for more than a decade. Recent research indicates that it when applied repeatedly as repetitive transcranial magnetic stimulation (rTMS) has an antidepressant effect. RTMS is based on the principle of electro-magnetism. An electromagnetic coil placed on the scalp produces a time-varying magnetic field, which gives rise to a current in the proximity of the cerebral cortex. Unlike electroconvulsive therapy (ECT) rTMS does normally not give rise to epileptic seizures and does not require anaesthesia. This review covers a critical summary of the literature on the subject. The results of recent placebo-controlled, randomized trials are promising. However, further investigations are required, before rTMS can be fully integrated in the antidepressant therapeutic armamentarium. PMID- 10827560 TI - [Placebo effect in the drug therapy of depression]. AB - This paper estimates the effect sizes of placebo treatment of depression from meta-analyses of antidepressive medicine, and examines methodological artifacts in antidepressive research. It is concluded that between one-half and two-thirds of the effects of antidepressive medicine are explained by placebo response with a mean difference between treatment and placebo groups around 25 percent. This modest effect size should probably be adjusted for methodological bias due to incomplete blindness and other sources of error favouring the medicine group. Some practical consequences of the fact that depression is a placebo sensitive disorder are outlined. PMID- 10827561 TI - [Refugee and immigrant children referred to a department of pediatric psychiatry during the years 1991-1994]. AB - The purpose of this study was to describe a group of 42 refugee- and immigrant children up to 14 years old compared to Danish controls in order to identify groups at risk and describe course, examination, treatment and contact. The investigation was based on case records. The only statistically significant results were higher frequencies of referral than expected regarding children from Lebanon and Somalia. However, the data indicate differences between the foreign and Danish groups as to greater problems of speech and language and less behavioural and emotional problems among the foreigners. No foreign children were offered foster homes. The attitudes of the parents towards referral, examination and treatment are more negative in the foreign group. The need for more research is emphasized. PMID- 10827562 TI - [The effect of patient-specialist-continuity in a surgical department. Evaluation of seven years of experiences]. AB - From 1990 through 1997 the working schedules for staff surgeons underwent gradual changes. The aim was to comply with patient demands to receive care from few surgeons during hospital care. To evaluate the effect, four controlled or cross sectional studies were conducted. Quality of care was assessed by surgical audit. Patient and surgeon satisfaction was examined with questionnaires and interviews. Changes in costs were evaluated by administrative data. The number of different specialists providing care was reduced by 50%, initially during inpatient care and later also during outpatient care. The patients experienced this change as very satisfying. Hospital stay was reduced, as was time used by specialists on out-of-hours duty. The quality of care did not change neither did the use of diagnostic tests. The specialists appraised the increased individual responsibility in patient care as satisfying. Initiatives should be taken to examine whether other ways of organising work for hospital employed doctors have similar effect. PMID- 10827563 TI - [Prevalence of depressive disorders among frail elderly, assessed by psychogeriatricians, general practitioners and geriatricians]. AB - The prevalence of depressive disorders (D) in 70 years of age or older frail elderly was studied. Two hundred and eleven (81%) recipients of Municipal Home Help Service (frail elderly) in a rural area of Jutland were included and screened by the Geriatric Depression Scale-15 (GDS). If participants had: a) a GDS-score > or = 5 points or/and b) a history of depression, further evaluation was given by: 1) general practitioners (GPs) (implicit criteria), 2) a psycho geriatrician (ICD-10 criteria), and 3) a geriatrician with the Hamilton Test for Depression. Prevalence rates for D among the evaluators: 15-18%. The diagnostic agreement was only fair. IN CONCLUSION: 18% of the studied population fulfilled the ICD-10 research criteria for depression. The GDS may help GPs in diagnosing depression among frail elderly. False GDS negatives, found among GDS negatives with a history of depression, should be evaluated thoroughly when identifying depression in the target group. PMID- 10827564 TI - [Endoscopic treatment of vesicoureteral reflux in children]. AB - In the five-year period 1990-95, 102 patients, 78 girls and 24 boys, aged ten weeks to 14 years were treated for grade III-V vesicoureteral reflux by the same surgeon using subureteric teflon injection--the STING procedure. Both primary, secondary and complex reflux were included. The reflux had disappeared in 90 of 133 refluxing ureters, the grade of reflux had diminished in 17 and were unchanged in 26 at the control three months after treatment. After re-STING in 22 reflux ureters, reflux disappeared in 13, was reduced in six and unchanged in three. Reappearance of low grade reflux was seen in eight of 88 ureters between the three months and one year control, but only one needed re-STING. Open surgery was performed in 13 patients (18 reflux ureters) because of unchanged reflux after STING, most of them due to complex reflux: duplex ureters, ectopic ureters, ureteroceles and paraureteral diverticula. It is concluded that most patients with vesicoureteral reflux can be successfully treated by using subureteric teflon injection. PMID- 10827565 TI - [When time stops... Psychiatric reactions to deportation orders of unaccompanied children and teenagers seeking asylum]. AB - Five cases of psychiatric disturbances in unaccompanied children who were refused political asylum are described. The children showed severe reactions with suicidal thoughts or psychosis, with a characteristic loss of perception of time during the long period of waiting for a decision. These children with severe reactions constituted a minority of the total who were refused asylum. PMID- 10827566 TI - [The type of hospital and results of hip alloplasty]. PMID- 10827567 TI - [Control of tuberculosis in Denmark]. PMID- 10827568 TI - [Otomycosis]. PMID- 10827569 TI - [Assertive community team--better than what?]. PMID- 10827570 TI - [Take your own responsibility when it comes to your health!]. PMID- 10827571 TI - An age limit for mammography? PMID- 10827572 TI - How to survive a hospital stay. PMID- 10827573 TI - A weight loss prescription for longer life. PMID- 10827574 TI - Hair loss treatments: weighing the pros and cons. PMID- 10827575 TI - My glaucoma has been hard to control, and now my ophthalmologist says I have something called pseudoexfoliation syndrome. What does this mean? PMID- 10827576 TI - Is calcium citrate better than other calcium supplements? PMID- 10827577 TI - Lesbian health care issues. Exploring options for expanding research and delivering care. PMID- 10827578 TI - C-sections recommended for HIV+ pregnant women. PMID- 10827579 TI - SCHIP: State Children's Health Insurance Program. AB - After the flurry of legislative activity in the late 1980's to expand Medicaid for pregnant women and children, the move from federalism to states' rights led to incremental health care program expansion. Despite the increased coverage in current programs, the number of uninsured women and children has increased since 1990. During 1997, 13.7 percent of all pregnant women (about 465,000) were uninsured (March of Dimes, 1999) as were 11.3 million children under the age of 18 (Children's Defense Fund, 1998). PMID- 10827580 TI - Charting in record time. AB - Nurses working in intrapartum settings have many responsibilities to ensure a safe and satisfying birth experience for the families in their care. Paperwork, while essential, often takes time away from providing supportive care for laboring women and their families. In striking a balance between responsibilities and obligations, nurses must play an active role in designing a charting system that allows essential and important data to be recorded in the least amount of time. PMID- 10827581 TI - Menopause, naturally. Exploring alternatives to traditional hormone replacement therapy. PMID- 10827582 TI - Harmonizing herbs. Managing menopause with help from Mother Earth. PMID- 10827583 TI - Building alternative families. Helping lesbian couples find the path to parenthood. PMID- 10827585 TI - MEDLINE revisited. PMID- 10827584 TI - Learning about genetic risk. Weighing the risk with the right to know. PMID- 10827586 TI - Nursing in the next millennium. Leadership skills for surviving and thriving. AB - Consider the following case study: The setting is an acute care facility with a large obstetrical service delivery (6,000 babies annually.) The year was 1989. A nurse leader had been appointed manager of Women's Services. Major job responsibilities included staffing, hiring, employee performance evaluations, developing policies and protocols, and managing daily operations issues. The reimbursement for the patient population was more than 50 percent fee for service. PMID- 10827587 TI - A time for dying. Working through end-of-life decisions. AB - "I think it's time," said Bruce, between sobs. I knew my husband meant it was time to help our son Roger die. Roger entered our family 18 years ago. He was a tiny (1 lb., 12 oz.) premature baby I met in the NICU. Because normal gestational age at birth is 38 to 41 weeks, Roger's gestational age of 28 weeks made him extremely vulnerable to many medical complications, such as chronic respiratory problems, intracranial hemorrhage, mental retardation, and seizures. He experienced all of these complications during his life. PMID- 10827588 TI - Nephrology nursing practice: more than technical expertise. AB - This paper argues that the role of the nephrology nurse goes far beyond the generally accepted parameters of technical expert to include the roles of care giver, advocate, educator, facilitator, mentor and referral agent. Patients with End Stage Renal Disease, who require long term dialysis, experience many stressors which they must cope with in order to achieve an acceptable quality of life. Nephrology nurses who understand the stress of living with dialysis, and who are familiar with the coping strategies a patient may use, are ideally placed to facilitate the process of adaptation from the patient and their families perspective. PMID- 10827589 TI - Total quality improvement assessment of the EPO co-ordinator role. AB - Total quality management or total quality improvement has developed and grown from the process of quality assurance. Its main features are, quality is a management led approach, top management participation is essential, it is a concept for preventing mistakes, not finding fault and correcting it. A client centred approach and teamwork are seen as key components. This paper explores the role of the Epo co-ordinator and identified what changes were required to improve the quality of the service. PMID- 10827590 TI - Preparing for the challenges of the millennium. AB - This paper describes a retrospective view of the challenges the two authors were faced with as Nurse managers when the haemodialysis unit was expanded from ten dialysis stations to thirty-one within a period of three years. This expansion involved changes to the environment and management strategies. There were four issues identified that helped to face these challenges and they are described in depth. The authors claim the skill of communication within the team and the proactive approach to change was the key to their success. PMID- 10827591 TI - The utilisation of shared governance to improve vascular access catheter care. AB - This article looks at the implementation of shared governance at directorate level and its role in improving vascular access catheter care. PMID- 10827592 TI - Malaria. A challenge for the haemodialysis and intensive care nurse. AB - Malaria is the world's most important tropical parasitic disease. In many developing countries, and Africa in particular, malaria exacts an enormous toll in lives, medical costs, and days of lost labour. Through the use of case studies, this paper explores the effects of malaria and the patient management by the haemodialysis and intensive care nurse. All malaria cases admitted to an intensive care unit, and treated by means of haemodialysis or continuous-veno veno-haemodiafiltration were analysed. Specific problems identified were impaired fluid and biochemical control, acid-base imbalance, disseminated intra vascular coagulation, adult respiratory distress syndrome, and haemodynamic instability. Guidelines were compiled for the nursing management of these complicated malaria cases. PMID- 10827593 TI - Algorithmic nursing protocol to provide optimum care post combined kidney/pancreas transplantation. AB - This paper explains the development of protocols to aid in the care of patients following a combined kidney/pancreas transplant. With adequate planning of care, medical and nursing problems can be detected at an early stage thus ensuring immediate and appropriated action. We have found protocols to be invaluable in the care of these patients and the one which we developed on fluids and electrolytes is outlined in this paper. PMID- 10827594 TI - Quality of water used for haemodialysis: bacterial and chemical parameters. I. Vorbeck-Meister et al. Nephrology, Dialysis and Transplantation, 1999 14 666-675. PMID- 10827595 TI - Exit-site infection prevention and treatment protocol. AB - Infections are universally known as one of the main causes of drop-out in peritoneal dialysis. Exit-site infections are often a problem, as they are difficult to treat, tend to become chronic and may lead to the development of continuity peritonitis, resulting in the need for removal of the peritoneal catheter. The purpose of this paper is to describe the application of an exit site infection prevention and treatment protocol. PMID- 10827597 TI - Relation between response to illness and compliance in haemodialysis patients. AB - The treatment schedule of the haemodialysis patient has many restrictions and the long-term haemodialysis patient's response to illness is characterised by various emotional reactions. Compliance with their medical regimens remains a significant problem which influences the progress of health and by extension quality of life. The preconceived idea that a patient's response to illness may influence his adherence to medical regimen led to this study. The main variables examined were the relationship between the patient's response to illness and compliance and the relationship between compliance and age, education and the length of time on haemodialysis. PMID- 10827596 TI - The patient with diabetic nephropathy in the hospital. AB - Diabetic nephropathy leads to chronic renal failure in 30-40% of diabetic patients. By offering primary health care and teaching patients with diabetic nephropathy about factors which influence their renal function, we can delay the renal deterioration and therefore the start of the renal replacement therapy. By learning to control their blood pressure, blood glucose and diet they will be fully aware of their health problem, avoid possible complications and last but not least they will possibly achieve more time. The nephrology nurse is capable of making a difference by offering advanced nursing care and being the patient's teacher and supporter. PMID- 10827598 TI - Stress, coping and quality of life among chronic haemodialysis patients. AB - This study identifies the most significant stressors perceived by individuals undergoing haemodialysis due to chronic renal failure, the coping methods used most often by patients, and their quality of life. 75 patients aged 18-65 years old, and on regular haemodialysis for at least for one year were studied. Results showed that patients perceived high levels of stress, and that psychosocial stressors are as problematic as the physiological ones. Patients used problem oriented coping methods more often than affective-oriented methods. Although their quality of life was satisfactory, patients were dissatisfied about their physical well being. PMID- 10827599 TI - Treatment decisions in paediatric dialysis: children's versus parental rights. AB - Treatment decisions in Paediatric Dialysis are often fraught with ethical questions about how to serve the best interests of children on dialysis, especially when parents refuse recommended treatment and opt for non-optimal interventions. This paper reviews the issues of children's rights, as guaranteed in the United Nations Covenant on Children's Rights and the Israel's Patients' Rights Law, and presents several case vignettes where conflict between children's rights for optimal treatment and parental desires arose. The paper concludes with policy recommendations to assure that children's expressed desires and medical needs are met, including the establishment of multidisciplinary committees to address the needs of children on dialysis and their families. PMID- 10827600 TI - An educational programme for persistent life-style changes in patients with chronic renal disease. AB - The aim of this study was to produce positive life-style changes in patients with chronic renal disease through a residential education programme about the medical aspects of chronic renal failure and the various treatment options in order to increase their physical capacity, emotional stability and general well-being. The programme was available to all categories of patients with chronic renal disease irrespective of treatment modality. Comparison of data pre and post course demonstrated improvements in physical activity, mental well-being and nutritional awareness. PMID- 10827602 TI - The nurse, the Internet, and consumer protection. PMID- 10827601 TI - Chronic disease: the challenge for nursing and midwifery. PMID- 10827603 TI - A critical social theory approach to nursing care of adolescents with diabetes. AB - This paper explores the viewpoint that, as a group, adolescents with diabetes are oppressed by health care professionals who use approaches to diabetes care delivery that are paternalistic in nature. These approaches can lead to negative health/diabetes outcomes in this age group. As inherent in oppressed group characteristics, adolescents with diabetes may not realize that they are being treated unfairly or that they have rights as patients. Nurses are in a unique position to advocate for these adolescents and to help them become advocates for themselves. PMID- 10827604 TI - Young Parents Project: a 21st-century nursing intervention. AB - The purpose of this exploratory, descriptive pilot study was to describe the usage of the Young Parents Project, a computer network offering health information and support, to adolescent mothers. Nine midwestern adolescent mothers were provided a computer so they could access the Young Parents Project in their homes. Participants gained access to the computer network 834 times for a total of 7,046 minutes during the year of the study. Adolescent mothers read and posted 402 electronic messages to one another and 110 electronic mail messages to advanced practice nurses. The following eight themes emerged when participants' electronic messages were analyzed using content analysis: introductions, emotional support, social support systems, infant sharing, infant issues, postpartum issues, finances and education, and computer support. The adolescent mothers' evaluation of the Young Parents Project was primarily positive, and the project served as a mechanism for providing health information and social support to adolescent mothers. PMID- 10827605 TI - Special needs children in the public schools: perceptions of school nurses and school teachers. AB - Schools are faced with the challenges presented by special needs children (SNC) because the law requires that they must provide educational opportunities to all children--those who have no handicapping conditions as well as those who do, no matter how severe those conditions. The need exists for adequately prepared health care professionals in the school setting. Using a convenience sample of school teachers and school nurses, this investigation focused on the perceptions of school teachers and nurses regarding the challenges and demands of having these children in the public school. Two surveys were conducted to study those perceptions. Quantitative and qualitative data analyses showed that the needs of both groups of providers--school nurses and school teachers--can be summed up in three categories: information dissemination, communication, and resource integration. Infrastructure development involves the establishment of an effective information management system, effective use of such a system in establishing communications between all participants, and adequate administrative support to facilitate the development of the school providers' sense of competence in the care of SNC. A well-planned and adequately supported program goes a long way toward changing people's attitudes toward the inclusion of SNC in the classroom. PMID- 10827606 TI - Sexuality and the adolescent with a physical disability: understandings and misunderstandings. AB - The purpose of this descriptive study was to examine the areas of sexual knowledge, sexual behavior, and beliefs about sexuality among adolescents with congenital physical disabilities. The sample consisted of 15 males and 14 females, ranging in age from 12 to 22 years. The Sexual Knowledge Interview Schedule (SKIS) was administered to all participants during face-to-face interviews. In addition, the participants were asked questions regarding their ability to engage in intimate relationships and their future childbearing potential. Overall, the findings suggested that these adolescents are uninformed or misinformed about general sexual knowledge, have many misconceptions about sexuality and their disability, and depend on health care professionals and parents for sex education. Implications of this research are discussed. PMID- 10827607 TI - Psychological support: are nurses appropriately prepared to deliver? PMID- 10827608 TI - Do relatives have a right to witness resuscitation? AB - A relative's right to witness resuscitation is the subject of considerable discussion and debate. This paper explores the presence of relatives in the resuscitation room from a moral and ethical perspective. The focus of discussion is essentially upon the principle of respect for autonomy vs. what appears to be the counter-argument, benevolent paternalism. It is concluded that recognition of a relative's right to witness resuscitation is dependent upon health care professionals' willingness to promote the principle of respect for autonomy. PMID- 10827609 TI - The importance of information giving for patients newly diagnosed with cancer: a review of the literature. AB - The importance of information giving has been recognized and promoted in political, ethical and professional arenas and this has ultimately resulted in the publication of a vast amount of literature relating to the subject. This literature review considers the functions of patient information and the factors influencing information seeking for patients who have been diagnosed with cancer. Methods of providing information are discussed and the nurse's role in information giving is examined. A number of core problems are highlighted in relation to information giving. These problems, and possible reasons for them, are subsequently critically analysed. Finally, the literature relating to information needs of patients with cancer is evaluated and five key needs identified. PMID- 10827610 TI - Mental health training needs of primary health care nurses. AB - The primary health care team (PHCT) has a critical role to play in the care of people with mental health problems. However, little attention has been paid to the mental health training and preparation required by primary health care nurses. This paper reports on a training needs assessment undertaken in one London health authority area. The findings indicate that primary health care nurses are undertaking a wide range and increasing volume of mental health work with little preparation and training. Specific training needs include time and space to identify training gaps; basic training in mental health, safe working practices and management of role boundaries; and multidisciplinary training. In addition, NHS trusts, primary care groups and health authorities need to clarify which aspects of nurses' mental health workloads are appropriate to their role and develop policies, procedures and training to support them in undertaking that role. PMID- 10827611 TI - Unwillingness to be violated: carers' experiences of caring for a person acting in a disturbing manner. An interview study. AB - Carers working in psychiatric care are sometimes exposed to insane, unpredictable and violent actions. In rare cases a patient appears to be resistant to all forms of pharmacological treatment. Fifteen carers (four registered nurses, 11 enrolled nurses) on a psychiatric ward in Sweden were interviewed about their experiences when caring for a person who acted in a disturbing manner. Narrative interviews were conducted and interpreted using a method inspired by Ricoeur. Four themes were formulated which describe the carers' uncertainty about the future, their inability to interpret the patient's disturbing behaviour and their own overall feeling of meaninglessness. The carers were of the opinion that the patient had the power and ruled the ward, which led to them feeling they were subjugated victims. The interviews also revealed the carers' recognition of forbidden feelings and actions and unknown negative sides. These results were interpreted and reflected on in the light of an ethical framework in order to achieve a deeper understanding of the text. This paper shows that an ethical perspective is important when searching for the meaning of caring for patients acting in a disturbing manner. The study raises the question: 'Is it possible to establish good when evil has dominion?'. PMID- 10827612 TI - Towards recovery: living in a home-like setting after the move from a hospital ward. AB - Six clients with a diagnosis of schizophrenia were interviewed about their experiences of their lives in a home-like setting, their key care provider and the care received. Their narratives of lived experiences in care were interpreted as living a process of health in the midst of severe mental illness, involving: 'becoming more', 'being disabled', 'comforting/conforming relationship', 'discomforting/unconfirming relationship' and 'caring about the caring relationship'. Fatigue and lack of strength influenced the lives of most clients considerably, and the stories were about problems and conflicts. Nevertheless, the clients seemed really to struggle to make communal life work, and there were experiences of increased competence and better self-confidence in most of the stories. On the whole care was described as good, and the process of health seemed to be supported by experiences of comfort and being confirmed in the client-care provider relationship. PMID- 10827613 TI - Narrative and narrative analysis. PMID- 10827614 TI - Reflective narrative and dementia care. AB - This study was an enquiry into how clinical supervision could enable a team of six community psychiatric nurses to provide improved patient-centred care. It also examined how feelings of burn-out, stress and tedium when caring for people with dementia could be relieved. The methodology used in the study analysed the process, enabling the findings to be studied. This is new paradigm research, requiring the researcher and participants to be equal and active coenquirers into the process facilitating the enquiry. Six community psychiatric nurses allowed their period of supervision to be recorded over six months in order to identify relevant issues of concern and resolution in their practice. It was anticipated that the experiences revealed during supervision would provide a means by which the nurses could develop their skill and knowledge and improve their care-giving. The findings suggest that the education of community psychiatric nurses needs to enable them to develop the ability to demonstrate emotional competence and to carry out self-questioning. PMID- 10827615 TI - Patients' experiences of having an underwater seal chest drain: a replication study. AB - The nursing management of patients who have a chest drain in situ has received little attention. The findings of a single small-scale study previously indicated that such patients' needs for information are not always fully met and pain control is inadequate. This small-scale study was replicated to produce broadly similar findings in a second hospital 2 years later. Despite some differences in the approach to management it was apparent that patients were still not well prepared pre-operatively and there was scope for improving pain control. All the patients reported considerable discomfort and pain of moderate to severe intensity. The type of pain described is typical of deep somatic/visceral pain. PMID- 10827616 TI - Patients' views on quality of care: age effects and identification of patient profiles. AB - Age differences among physically ill inpatients, regarding their evaluation of the subjective importance of various care conditions, as well as their actual evaluations of these conditions, were explored. Subjective importance ratings were found to be unrelated to age. Patients' perceptions of the care they actually received indicated increasingly more positive evaluations with increasing age. Profiles of more and less satisfied patients were obtained using cluster analysis. Lowest satisfaction was reported by younger and well-educated women with a lower sense of coherence (as measured by Antonovsky's (1987) version of the Sense of Coherence Questionnaire) who stayed for a short period of time in hospital. It was concluded that caregivers need to focus on individual patient characteristics when looking at quality of care from a patient perspective. Generalizations based on age alone tend to be misleading. PMID- 10827617 TI - Life course of people with coronary artery disease. AB - The purpose of this study was to describe the life course of coronary artery disease patients from their own perspective at the onset of coronary artery disease and during both the in-patient period and the one-year period of aftercare. Nineteen people selected from a population of 200 patients who had undergone either coronary artery bypass surgery or percutaneous transluminal coronary angioplasty were available for thematic interviews. Interviews were conducted one year after the treatment in the subject's homes. Analyses revealed two distinct types of life course; accepting and progressive, as opposed to non accepting and regressive. Participants who represented an accepting and progressive life course achieved a better level of rehabilitation than those with a non-accepting and regressive life course. If health care personnel are able to identify the problems related to a non-accepting, regressive life course, they will be better able to support patients' individual life course planning. According to the present findings, a rehabilitation programme is particularly needed for patients with acute onset of coronary artery disease at a relatively early age, disruption of an active working career, financial problems, dissatisfaction with outcome of treatment, family problems and a dismal view of the future. The findings challenge health care personnel to listen to coronary patients' own experiences. PMID- 10827618 TI - Women living with type II diabetes: the intrusion of illness. AB - This study is part of a larger research project which aimed to create an understanding of how people with type II diabetes incorporated chronic illness into their lives. We aimed to find wellness in the context of a chronic illness. The participatory action-orientated research (PAR) approach is consumer based and managed, and six women with diabetes and the research team met for eight two-hour sessions in August and September 1998. The objectives were to contribute to understanding of the health of adults who live with chronic illness by providing greater insight and understanding into the worlds of people who live with a chronic illness and allowing the voices of people with diabetes to be clearly heard in relation to their health. There was a wellness theme around taking time out, but considering the other dominant negative experiences, this study can only be considered as an attempt to find wellness in the context of chronic illness. The PAR process, built upon sharing, listening and reconstructing stories, worked toward the women being heard and having a voice for the first time. Listening and acting upon the voices of the women has implications for the practice of health care professionals. PMID- 10827619 TI - 'It's not the same as him being at home': creating caring partnerships following nursing home placement. AB - Admission to a nursing home is a major life event for both older people and family carers. Despite a policy of community care in both the UK and the US, entry to nursing homes will be an increasingly common event. Family carers are often the key decision makers but little is known about their experience of placement, especially adjustment after the event. Antagonistic relationships can often develop between staff and relatives, as the former seek to take over care and the latter to develop new roles. There is a need to create a partnership between staff and family so that the care of the older person is improved and the carers' need to remain involved is acknowledged. PMID- 10827620 TI - Determining older people's need for registered nursing in continuing healthcare: the contribution of the Royal College of Nursing's Older People Assessment Tool. AB - This paper presents an overview of the work of the Royal College of Nursing (RCN) in developing an assessment tool to determine the need for registered nursing by older people in continuing care. In the past, healthcare has been provided free and social care 'means-tested'. The erosion of the NHS's responsibility for continuing care provision through the development of eligibility criteria has resulted in the majority of older people having their care services means-tested, including their nursing care. The RCN has argued that older people have inter related health and social care needs, that nursing care is an integral component of healthcare provision and that therefore older people should not have to pay for nursing care. This paper describes the work undertaken by the RCN to develop a tool for use in assessing older people's need for registered nursing--'The RCN Nursing Older People Assessment Tool'. The underpinning theoretical framework is outlined and the process of constructing the tool is described. In addition, the stages of assessment using the tool are outlined. A pilot study in seven nursing homes managed by three different companies was undertaken to test the reliability and acceptability of the tool. 178 paired assessments were completed (89% of the intended sample). The results of the pilot study demonstrated that the tool has a high degree of reliability and acceptability. PMID- 10827621 TI - The development of a collaborative stroke training programme for nurses. AB - The effectiveness of multidisciplinary stroke rehabilitation may be enhanced by nurses and therapists adopting a single consistent approach to the positioning and mobilizing of patients. As patients can spend as little as 4% of the waking day receiving 'therapy' there is considerable potential for a more dynamic nursing intervention, which may contribute to improving patient care. We aimed to investigate whether physiotherapists could step back from direct patient treatment in order to participate in a structured training programme for nurses involved with patients recovering from stroke on an established elderly care rehabilitation ward in a district general hospital. Qualitative methods were used within a participatory action research framework to describe the development process and content of the training programme. Nursing staff, physiotherapists and their respective managers were interviewed to identify perceived training needs. This informed the structure and content of the training course and allowed insight into interprofessional working. PMID- 10827622 TI - The decision to resuscitate: older people's views. AB - The aim of this study was to explore how a healthy sample of the older population feel about resuscitation and the decision not to resuscitate certain patients. Their views were sought on whether a Do Not Resuscitate decision is appropriate in certain circumstances, whom they think should be involved in the decision, whether they think patients should be consulted, and if they would like to be involved in the decision themselves. Two focus groups were held in a day care setting in order to collect data, and a thematic analysis was conducted. Participants thought that a Do Not Resuscitate decision should be discussed with patients and also with relatives if appropriate. However, there was ambivalence about whether individuals would like to be involved personally in such a decision because of the anxiety this would produce. PMID- 10827624 TI - Spearman's rank correlation test. PMID- 10827623 TI - Needs of Chinese families with a relative in a critical care unit. PMID- 10827625 TI - Clinical governance; evaluation and intuition. PMID- 10827626 TI - Nursing workload: an unquantifiable entity. AB - BACKGROUND: In the increasingly prominent drives towards cost containment and efficiency nursing is under pressure to justify its value to healthcare. On the surface nursing workload assessment methods would appear to offer a means to quantify the nursing input to patient care. AIMS: To examine the validity of nursing workload methods with respect to nursing practice and nursing knowledge. FINDINGS: Previous studies and the conclusions of subject reviews have cast doubt on the reliability and validity of most methods and indicate that they have no clear theoretical base. CONCLUSION: It appears that nursing is more concerned with knowledge processing and nurses' intentions than just with the activities of caring. It is suggested that patients control the time element of nursing care. The need for nursing workload information, despite its validity problems, is discussed and a pragmatic solution is suggested. PMID- 10827627 TI - Quality management within a cost-constrained service. AB - AIM: To examine the meaning of quality and ways in which quality can be improved despite budgetary limitations. KEY ISSUES: Definitions of quality are discussed, comparing the development from industrial concepts of quality to that contained within the Department of Health's A First CLASS SERVICE: The relationship of cost and quality is examined and the effects of changing treatments on healthcare costs explored. Clinical error is identified as a cause of increased cost through failure to maintain quality. A variety of quality tools are discussed in identifying ways in which quality can be improved within a cost-limited service. Thrombolysis is used as an exemplar and the effects on nursing discussed. CONCLUSION: There is research evidence which demonstrates that some areas of quality are amenable to improvement despite budgetary constraints. PMID- 10827628 TI - A modernized mental health service: the role of care pathways. AB - AIM: This paper reviews the impact of the UK governments' strategy and structures to improve the quality of mental health care. The possible role of care pathways within this modernization agenda is discussed. BACKGROUND: The governments' emphasis upon reducing clinical variation and cost effective treatments complements both the historical and structural elements of care pathways. This paper is a reflection on some of the issues within this debate. METHODS: The paper reviews the latest publications from government sources and draws on the authors' research and critical analysis of the care pathway literature. FINDINGS: The government has made an explicit movement towards structuring care processes within the newly formed National Service Framework. However, there are many problems in following standardized care processes such as professional antagonism and a relative dearth of evidence-based practice. CONCLUSION: Further research is required to examine the wider impact of government initiatives on the work force. Care pathways may provide the structure for this research and development activity. PMID- 10827629 TI - Promoting quality of care for older people: developing positive attitudes to working with older people. AB - AIMS: Managers and administrators have a key strategic role and responsibility for the way the care of older people is delivered within health and social care services, since the decisions made and directions taken at this level have a direct influence on services delivered. This article provides an outline of the context in which the services provided for older people have emerged, especially within health care, and offers strategies for the way forward. BACKGROUND: The standard of care received by older people is high on the agenda in contemporary health care. Frequent reports have questioned this quality over the years, yet concerns still remain. A key factor influencing quality of care received by older people, can be attributed to the persistence and perpetuation of ageist attitudes held by society and those working within health and social care settings at all levels. METHODS: Drawing upon a range of literature, an overview of those factors that seem to be attributable to the development of contemporary attitudes and perceptions about older people and their care are reviewed, focusing particularly upon health and social care workers, especially nurses. FINDINGS: The role of education in particular is explored as this has a key role in influencing attitudes towards caring for older people and will impact directly on the way in which all care and services develop. CONCLUSION: A number of areas are outlined for future development and research that aim to address and serve the needs of older people, and which could be supported at a managerial and administrative level to promote positive attitudes. PMID- 10827630 TI - Implementing smoking policies within trusts: nurses' perceptions and views of effectiveness and implications. AB - AIM: This study set out to explore nurses' views and perceptions of their hospital Trust's smoking policy, and the effects that it has had on both staff and patients. BACKGROUND: Although soliciting and considering the views and perceptions of staff during smoking policy development increases the likelihood of effective implementation, such activity does not appear to be routinely undertaken within the NHS. METHODS: Semistructured interviews were conducted with seven surgical nurses working within one district general hospital in the UK. RESULTS: While all of the subjects acknowledged the value of there being a smoking policy, they felt that their Trust's policy placed nurses in a powerless and vulnerable position, with little support available to help effectively address the problems that smoking raised for them and their patients. CONCLUSIONS: NHS Trusts that wish to develop and implement effective smoking policies should engage their staff throughout the process, and not underestimate the positive contribution that they all can make. PMID- 10827631 TI - Promoting evidence-based practice: managing change in the assessment of pressure damage risk. AB - AIMS: This study set out to facilitate the development of evidence-based practice in the assessment of pressure damage risk to patients within a large acute hospital. BACKGROUND: The importance of nursing practice being based on the best available evidence is emphasized in recent health policy. Meeting this objective is not easy as both individual and organizational factors create barriers to the implementation of research findings and the achievement of change. METHODS: The study was based on an action research model. It comprised three stages: a review of the research evidence; a survey of qualified nurses' knowledge of risk assessment of pressure damage and an audit of record keeping, and a multifaceted approach to achieving change in which researchers, managers, practitioners and clinical nurse specialists worked together collaboratively. FINDINGS: The findings from the survey and audit indicated a shortfall in nurses' knowledge of risk assessment of pressure damage and in their record keeping. The researchers, with the help of the clinical nurse specialist, built upon these findings by assisting practitioners and managers to take ownership of the need to base practice on the appropriate evidence. CONCLUSIONS: Achieving evidence-based practice is a complex undertaking that requires the development of an evaluative culture and a commitment by practitioners and managers to change practice. Researchers can play a valuable role in facilitating this process. PMID- 10827632 TI - On the receiving end of health care. PMID- 10827633 TI - A guide to revising the postanesthesia care unit documentation record. AB - The environment of the PACU is changing rapidly, as are the standards, protocols, and requirements of documentation. Nursing care activities must be captured in a timely, accurate, and efficient manner that reflects the standards of care and quality practice, while reducing the amount of actual writing required. This article reviews the available literature and defines a process to guide one through the revision of a documentation record. PMID- 10827634 TI - Adapting charting by exception to the perianesthesia setting. AB - Many acute care settings are experiencing a shift toward ambulatory patient care and are being pressured to decrease patient length of stay. Meanwhile, there is increased pressure to remain patient centered and to become more customer oriented. This article describes how the authors adapted charting-by-exception principles to develop a single ambulatory surgery nursing record for use from admission to discharge. The authors' goals were to condense documentation, to decrease time spent on documentation, and to diminish the variance in quality of documentation. This article also details how the new nursing record measures achievement of patient outcomes, with a focus of promoting street readiness. PMID- 10827635 TI - Ambulatory anesthesia and surgery in America: a historical background and recent innovations. AB - This article describes the history of ambulatory surgery and anesthesia in the United States. The innovations in surgical instrumentation and techniques, newer anesthesia drugs and techniques, as well as insurance provider changes that have led to an increase in the numbers and types of surgeries performed in outpatient settings in the United States, are detailed. PMID- 10827636 TI - When a minute seems like a millennium. AB - Surgery is a stressful experience for patients and their families. The increased use of ambulatory surgical services, coupled with changes in health care economics, requires nurses to design innovative ways of addressing the emotional needs of patients and families, while not compromising standards of care or becoming frustrated with the fast-paced world of ambulatory surgery. This article presents ways to help patients and their families throughout the preoperative experience. PMID- 10827637 TI - Antihypertensive agents. AB - Fifty million Americans suffer from hypertension. During the perioperative period, hypertension is a frequent finding owing to a variety of mechanisms including anxiety, pain, and preexisting conditions. This article explores the pharmaceutical agents commonly used in the treatment of hypertension. Particular attention is directed to drugs administered in the perioperative setting and complications that can be observed. PMID- 10827638 TI - What can the postanesthesia care unit manager do to decrease costs in the postanesthesia care unit? AB - The economic structure of the PACU dictates whether a cost-reducing intervention (e.g., reducing the length of time patients stay in the PACU) is likely to decrease hospital costs. Cost-reducing interventions, such as changes in medical practice patterns (e.g., to reduce PACU length of stay), only impact variable costs. How PACU nurses are paid (e.g., salaried v hourly) affects which strategies to decrease PACU staffing costs will actually save money. For example, decreases in PACU labor costs resulting from increases in the number of patients that bypass the PACU vary depending on how the staff is compensated. The choice of anesthetic drugs and the elimination of low morbidity side effects of anesthesia, such as postoperative nausea, are likely to have little effect on the peak numbers of patients in a PACU and PACU staffing costs. Because the major determinant of labor productivity in the PACU is hour-to-hour and day-to-day variability in the timing of admissions from the operating room, a more even inflow of patients into the PACU could be attained by appropriate sequencing of cases in the operating room suite (e.g., have long cases scheduled at the beginning of the day). However, this mathematically proven solution may not be desirable. Surgeons, for example, may not want to lose control over the order of their cases. Guidelines for analysis of past daily peak numbers of patients are provided that will provide data to predict the minimum adequate number of nurses needed. Though many managers already do this manually on an ad hoc basis statistical methods summarized in this article may increase the accuracy. PMID- 10827639 TI - Pruritus and urticaria. AB - Pruritus and urticaria frequently occur during the perianethesia period. Knowledge of the physiology of these symptoms can assist the perianesthesia nurse to identify the appropriate nursing and medical interventions. PMID- 10827640 TI - The energy business. AB - Looking at change as a great opportunity for growth puts a different perspective on the response both managers and employees exhibit. To rekindle motivation and energy in the workplace, the very mind-set of each individual must change. The potential for growth rather than fear of loss creates energy and excitement. It can ultimately lead to a workplace where there is joy and a strong sense of accomplishment. PMID- 10827641 TI - The life of a new long stay patient: a reflexive experience. AB - The care received by one patient in an acute psychiatric ward setting is described and analyzed by examining the relevant literature and the patient's perspective through a therapeutic relationship with a psychiatric nursing student. The patient is described as the new long stay patient and the reasons for the existence of this group are discussed. The paper concludes by stating the need to provide for those patients who are inappropriately occupying acute beds because there is no available alternative. This is found to cause problems for the individual, for psychiatric services and for nursing care. PMID- 10827642 TI - Creating alliances: the views of users and carers on the education and training needs of community mental health nurses. AB - The Sussex Education and Training Consortium commissioned a development project to identify the education and training needs of Community Mental Health Nurses (CMHNs) working with people with severe mental illness. The concerns of various stakeholding groups were identified using a qualitative methodology, Responsive Evaluation. This paper reports the views of 52 service users and representatives and 24 carers and career support workers as expressed in semi-structured interviews and focused discussion groups. Users and carers were able to identify specific areas that require attention in the education and training of CMHNs. Users want CMHNs to have wide knowledge of mental illness, medication, therapies and services and the understanding and confidence to work in an empowering way. They also want CMHNs to systematically involve users and their carers in care planning and to make more explicit any communication with other professionals and agencies. Carers want more involvement and information and CMHNs made more aware of the role that carers play and the specific problems they face. Users and their organisations would like significant involvement in the provision of CMHN education and training at all levels. Developments to CMHN education and training are suggested with recommendations for research and evaluation. PMID- 10827643 TI - The psychiatric nurse and the 'appropriate adult'. AB - This paper identifies a role for the mental health nurse in the 'Appropriate Adult Scheme'. It discusses how the unique skills of the mental health nurse could be utilized in this role but also the problems and objections associated with this potential development. PMID- 10827644 TI - Empathy: a crucial component of the helping relationship. AB - The literature reviewed in this paper substantiates the widely held view that empathy is crucial to all forms of helping relationships. While most studies cited are more than a decade old, the relationship between empathy and helping remains unchallenged in the 1990s. Additionally, while there is confusion about whether empathy is a personality dimension, an experienced emotion, or an observable skill, it is shown that empathy involves an ability to communicate an understanding of a client's world. Finally, a definition of empathy considered to be relevant to clinical nursing is introduced, which includes the need to understand client's distress, and to provide supportive interpersonal communication. It is argued that there is a need to revisit the role of empathy in the context of current health care delivery. PMID- 10827645 TI - Effects of systematic clinical supervision on psychiatric nurses' sense of coherence, creativity, work-related strain, job satisfaction and view of the effects from clinical supervision: a pre-post test design. AB - There are few investigations of the type and the outcome of interventions aimed at supporting nurses caring for psychiatric patients. Therefore a prepost-test design study was used in which 22 psychiatric nurses, on a general psychiatric ward were examined before, during and after one year of systematic clinical supervision combined with supervised documented, planned, individualized care. The methods used were the Sense of Coherence scale (SOC), the Creative Climate Questionnaire (CCQ), the Work-Related Strain Inventory and 34 statements from the Satisfaction with Nursing Care and Work Questionnaire (SNCW). In addition 14 statements were developed to evaluate the nurses' view of the effects from clinical supervision. The baseline values for the CCQ indicated a stagnant organization and a high score in the conflict dimension indicated personal and emotional tensions within the organization. The intervention led to a significantly increased creative and innovative climate in the dimensions for trust, idea time and reduced conflicts. However, the organizational climate remained stagnant. The nurses' view of the effects from clinical supervision also increased significantly. There were no significant changes in the nurses' SNCW, WRSI or SOC score. The result of the correlation analysis indicated that a strong sense of coherence was related to low work-related strain but not to unsatisfactory working conditions/milieu. The results gave some support to the idea that systematic clinical supervision and supervised nursing care plans constitute one type of support strategy that improves creativity and the organizational climate. It may, not, however, buffer for interpersonal problems. Further research is required to explore the need for effects of various support systems depending on the circumstances in the organization. PMID- 10827646 TI - Effective health care for people with learning disabilities: a formal carers' perspective. AB - This paper explores the perceptions and experiences of formal carers, working in three different geographical locations in the south of England, in accessing primary health care for people with learning disabilities. The sample included six multidisciplinary, community learning disability teams and 11 managers of small homes for residents who had learning disabilities. Data were collected through focus group and individual semistructured interviews. Levels of collaboration between primary health care groups and the community learning disability teams and the managers of small homes were assessed. Questions were focused upon the importance of primary health care and the key care provider within the community setting. Although some good experiences were recorded, the overall levels of collaboration were poor. Health care was perceived as very important by all of the respondents. The general practitioner (GP) as the coordinator of health care was perceived as the normal model within our health care system but the majority of respondents questioned the effectiveness in relation to their own experiences of accessing health care for people with learning disabilities. The relocation of this role to the learning disability nurse was perceived as a possible solution. PMID- 10827647 TI - A report of the development of a Chinese and British education initiative. AB - A significant event in the development of psychiatric and mental health nursing in China was conducted at An Ding Hospital, Beijing from May 5th to 9th 1999. The Peking Union Medical College, School of Nursing and An Ding Psychiatric Hospital hosted a series of workshops, conducted by Professor Philip Burnard (University of Wales), Dr David Arthur (Hong Kong Polytechnic University) and Madam Wang Zhiying (Director of the Nursing Department An Ding Hospital) for approximately 100 nurses from various regions of China. PMID- 10827648 TI - Virtual insanity: imag(en)ing illness in brain scans of schizophrenic people. PMID- 10827649 TI - Nurse-patient interaction: perspectives from the operating department. PMID- 10827650 TI - The next millennium. PMID- 10827651 TI - Management of a neonatal wound on a newborn infant. PMID- 10827652 TI - A non-comparative multi-centre clinical evaluation of a new hydropolymer adhesive dressing. AB - This study evaluates the performance of a new hydropolymer adhesive dressing in the treatment of chronic or acute wounds, in the presence of blood and/or low levels of exudate, in 74 patients, in a five-site multi-centre non-comparative clinical trial of four weeks' duration. The primary efficacy variable was defined as the incidence of central island dressing adherence to the wound bed in the final stages of healing. The hydropolymer dressing performed extremely well with 98.4% (240 out of 244) of the dressings not adhering to the wound bed. PMID- 10827653 TI - Evaluation of a range of hospital replacement mattresses. AB - A combination of physical measurement and clinical testing was used to evaluate a range of pressure-reducing replacement mattresses, which had met tender specifications, in order to reduce the number available for selection in the purchasing process. Factors considered included mattress price and expected life span. The results presented have supported a purchasing decision to replace mattresses but care should be taken in adopting these findings in a different context from that of a district general hospital. Optimum performance was found in one mattress replacement (Pentaflex). PMID- 10827654 TI - Infection control properties of some wound dressings. AB - The ability of some wound dressings to sequester and retain micro-organisms associated with wound fluid is perceived to provide beneficial properties regarding infection control. This study used an in vitro model to investigate and compare such properties in a range of fibrous absorbent dressings (alginate, hydrofibre and hydrophobic). Dressings were challenged with a simulated wound fluid containing common wound pathogens (Staphylococcus aureus or Pseudomonas aeruginosa). Bacterial sequestering and binding levels were monitored over time. A hydrofibre dressing and two calcium alginate dressings were shown to effectively sequester challenge organisms from a simulated wound fluid. However, the hydrophobic and hydrofibre dressings produced statistically significant results in their ability to adsorb and retain challenge organisms (p < 0.05). These investigations have demonstrated that a hydrofibre dressing effectively sequesters and retains micro-organisms upon exposure to simulated wound fluid, and may therefore provide a passive mechanism for reducing the microbial load in wounds and in the surrounding environment. Further in vivo studies are required to investigate these dressing properties. PMID- 10827655 TI - Hypergranulation tissue. PMID- 10827656 TI - Management of scarring. PMID- 10827657 TI - Measurement of breast skin viscoelasticity and a pilot study on the potential radioprotective effect of a zinc-based cream. AB - Radiation-induced late skin effects were studied in patients with breast cancer in relation to different protocols of fractionated radiotherapy in three different medical centres, in Israel, the UK and the USA. The mechanical properties of skin were evaluated in breasts of healthy volunteers, and non irradiated and irradiated breasts of patients, using a newly developed viscoelasticity skin analyser (VESA). The increase of the dose of radiation per fraction was found to have more impact on the development of radiation-induced late skin effects than the elevation of the total dose given. In addition, a pilot study on the possible radioprotective effect of external application of a cream containing zinc oxide on radiation-induced early skin changes in patients with breast cancer was initiated. Non-invasive measurement of trace elements and zinc pharmacokinetics in the skin of healthy controls following the application of the zinc oxide cream were performed by unique diagnostic X-ray spectrometry (DXS). Application of the cream, followed by thorough skin cleansing, significantly increased the amount of residual zinc in the skin, but continuous daily treatment did not cause further build-up of the dermal zinc level. The radioprotective effect of the zinc oxide cream on the skin is now being studied. PMID- 10827658 TI - Clinical evaluation of a new pressure-relieving mattress. AB - In introducing a new mattress into practice it is important to evaluate its performance from both the patient's and nurse's perspective. A total of 20 patients admitted to the integrated medical unit were recruited and nursed on a new pressure-relieving mattress offering two modes of pressure relief (Duo). All patients had been recently admitted to hospital and were considered to be 'at very high risk' of developing a pressure ulcer, with an initial Waterlow score > 20. Data were collected on admission, mid-point (three to four days) and study exit (seven days). The primary objective was to investigate the condition of patients' skin and the mode of choice for pressure relief. The secondary objective was to investigate reasons for nurses' and patients' choices. Structured interviews were conducted with the patient, ward nurse and research nurse whenever possible. Nineteen patients maintained or improved their skin condition during the evaluation. PMID- 10827659 TI - Topical insulin in wound healing: a randomised, double-blind, placebo-controlled trial. AB - Two studies were carried out to assess the relative roles of insulin and zinc in the acceleration of wound healing. In the first study, six diabetic and five non diabetic human volunteers had two uniform cuts created, one on each forearm. One forearm wound was treated with topical regular insulin (Iletin-II) and the other with normal saline four times a day until healed. Treatment was double-blind and forearms were assigned randomly. The wounds treated with insulin healed 2.4 +/- 0.8 days faster than the wounds treated with saline (P < 0.001 by paired t-test). Zinc is used to crystallise insulin. When wounds are treated with insulin, they are therefore also being treated with zinc. If insulin accelerates wound healing, it is not clear if the increase in the rate of healing would be due to insulin (a known growth factor), the zinc it contains, or a combination of the two. The second study used a randomised, double-blind, placebo-controlled design to compare the efficacy of insulin with that of a solution containing the same amount of zinc in accelerating the healing of standardised wounds in rats and humans. Although these pilot investigations did not have the power to define the relative roles of insulin and zinc with accuracy, the results suggest that zinc does play a role in the wound healing process. It is concluded that topical insulin accelerates wound healing in humans. More importantly, however, this study describes a method of creating uniform wounds in humans acceptable to an institutional review board, thus solving one of the major impediments to the scientific evaluation of human wound healing. PMID- 10827660 TI - Orthotic intervention in patients with diabetic foot ulceration. PMID- 10827661 TI - The palliative management of fungating malignant wounds. AB - This study focused on the palliative management of fungating malignant wounds and individual experiences of living with such a wound. Dressings were evaluated for the ability to contain these wounds and reduce their impact on daily life. The project extended to collaboration with industry for the development and evaluation of dressings designed to meet patient needs. A longitudinal multiple case study design was adopted. The methodology evolved through three principal phases: quasi-experimental design; emergent collaborative design; and emergent theory-driven evaluation. The radical departure from the initial approach was in response to the methodological problems encountered in a study of individuals with uncontrolled disease. A non-probability sampling plan was adopted, mainly because of the lack of homogeneity in the patient population; 45 participants were included. The length of time patients remained in the study depended on how long they lived. This ranged from a few days to more than two years. A sampling plan was, however, adopted for the data collection. The study had a dual focus: methodology, and the generation of explanations for dressing performance and the management of fungating wounds. The methodological aspect included development of the Teler system as a method of measuring dressing performance against goals of optimal practice in fungating wound management. The second component was a system of reasoning developed as an analytical strategy for abstracting general issues from individual case study data in order to construct explanations. Theory was used to generalize beyond the individual cases. Two forms of explanation for fungating wound management were constructed. These included explanations of individual experiences of living with such a wound and knowledge of the elements of fungating wound management. The impact on the individual was explained in terms of the stigma attached to public disability and a revulsion in society for uncontrolled body fluids. A pivotal relationship emerged between exudate and other wound management problems, including psychosocial aspects. A final critical explanation was developed for the qualification of the theory of moist wound healing to explain the phenomenon of exudate management in fungating and, possibly, other exuding chronic wounds. PMID- 10827662 TI - The use of silicone gel sheeting in the management of hypertrophic and keloid scars. PMID- 10827663 TI - The prevention of pressure ulcers in the operating department. PMID- 10827664 TI - The use of case reports in wound care. PMID- 10827665 TI - Comparison of a new foam mattress with the standard hospital mattress. AB - This study evaluates pressure ulcer incidence rates and comfort perceptions in 100 subjects admitted to a district general hospital. Subjects were divided into two groups, Group A using the standard hospital mattress (Transfoam), which had been in use in the hospital for three years, and Group B using the study mattress (Transfoamwave), which was new at the beginning of the trial. Due to the low observed incidence of pressure ulcers, it has not been possible to determine whether there is a difference in the clinical performance of the mattresses. Both appear to provide similar levels of comfort. Seat cushions were provided for each trial subject but staff failed to make good use of them, which suggests that more education is required in this area. PMID- 10827666 TI - The use of a hydrofibre dressing in wound management. PMID- 10827667 TI - Pressure-induced skin lesions in pigs: reperfusion injury and the effects of vitamin E. AB - The pathogenesis of the development of pressure ulcers is still unclear. The aim of this study was to investigate the role of ischaemia and reperfusion in pressure-induced tissue necrosis in the trochanteric region in pigs. Pressure application was achieved with a newly developed computer-controlled pressure device. Histological examination showed damage in the subcutis and muscle tissue comparable with inflammation, extending in a vascular pattern beyond the area of pressure application. Electron-microscopic studies revealed neutrophil adherence to the capillary endothelium, which showed signs of injury. These observations were manifest two hours after the cessation of pressure. Pre-treatment with 500 mg vitamin E per day resulted in significantly less tissue damage compared with untreated animals. Pressure alone caused a significant decrease in reduced glutathione and total glutathione, suggesting oxidative stress. After pressure release there was a significant increase in hydrogen peroxide concentration, suggesting a decreased antioxidant protection. After pre-treatment with vitamin E, however, there was no increase of hydrogen peroxide. It is concluded that the early signs of necrosis after pressure application are concordant with typical ischaemia-reperfusion damage and this can be prevented in part by treatment with vitamin E. Prophylactic administration of vitamin E may influence the occurrence of pressure ulcers in humans undergoing elective surgery. PMID- 10827668 TI - The role of the specialist nurse in diabetic foot ulceration. PMID- 10827669 TI - A non-healing leg ulcer associated with malignant lymphoma. PMID- 10827670 TI - Should the exception be lifted that disallows ESRD patients from enrolling in managed care programs? PMID- 10827671 TI - Providing dialysis services for patients in a skilled nursing facility. PMID- 10827673 TI - NKF president sheds light on status of DOQI nutrition guidelines. PMID- 10827672 TI - A reduction in the nursing labor market and its impact on the renal industry. AB - Nurses, especially those with specialized training such as nephrology, are looking more and more endangered. A cursory review of the classified ads supports the likelihood of another nursing shortage with the reappearance of key words such as, "relocation assistance," "incentive pay," and "hiring bonus." It is now commonplace to see hiring bonuses ranging from $3,000-$6,000. This article will investigate the definition of occupational shortages, possible causes of the nursing shortage, future implications, and strategies to be considered. PMID- 10827674 TI - Preparing dialysis equipment for Y2K: organization is the key. PMID- 10827675 TI - Administering independently. PMID- 10827676 TI - Team rehabilitation: different perspectives bridge the gap for better patient centered care. PMID- 10827677 TI - The forum of ESRD networks: past, present and future. AB - The increasing visibility and credibility of the Forum of ESRD Networks in the national ESRD landscape is due, in large part, to the Forum's longstanding and unwavering advocacy for improved ESRD patient outcomes through the application of continuous quality improvement methodologies and the development of a data infrastructure which encompasses a universal patient sample. This advocacy is untainted by the agenda of any single professional constituency, and its success is limited only by the commitment that all stakeholders (payers, providers, and patients) have to the process. The data infrastructure, SIMS-VISION, is almost a reality, and has the potential to significantly improve the quality of care through the provision of provider-specific profiles to drive quality improvement. The application of quality improvement principles themselves will take longer, and requires the commitment of facility medical directors, whom the Forum hopes to reach through its collaborative education project with the RPA. Through its clearinghouse activities, the Forum hopes to foster evidence-based medicine, increasing provider awareness of clinical practice guidelines and other literature which may improve the quality of patient care. The Forum's strategic plans for achieving its goals closely parallel the recommendations of the President's Advisory Commission on Consumer Protection and Quality in the Health Care Industry, and emphasize not only the information infrastructure, quality measuring/reporting, and evidence-based medicine, but also enhancing patient participation, building partnerships, and facilitating education and research. The Forum's strategic plan, as reconfigured according to the President's Advisory Commission domains, has been endorsed by the Renal Coalition as the national renal quality agenda, and the Coalition's constituent organizations are exploring projects and funding sources to achieve some of these goals as well. PMID- 10827678 TI - Workplace study to advance practice. PMID- 10827679 TI - Nurse-teacher roles--2. Balancing provision of education and service needs--is there a way? PMID- 10827680 TI - Critical appraisal. 8. Appraising qualitative research. PMID- 10827681 TI - Clinical governance. 6. Effective staff--2. PMID- 10827682 TI - Evaluating the quality of the learning experience. 1. Quality in learning. PMID- 10827683 TI - Your staff is watching you. PMID- 10827684 TI - Developing IT relief for chronic illness. AB - Read about emerging models of care delivery for the chronically ill and how information technology supports them and the search for cures. PMID- 10827685 TI - Clinical practice guidelines: consider, adopt, or do your own? AB - Learn how to find, adapt, and monitor clinical practice guidelines, which are outlined in four new JCAHO standards. PMID- 10827686 TI - What's your liability in a survey? AB - You have liability exposure for regulatory issues when federal, state, and local officials, as well as other organization, such as JCAHO, survey your facility. Answer the questions about the following survey scenario below, then check your answers at www.nursingmanagement.com. PMID- 10827687 TI - Nurse managers flex their budgetary might. AB - A survey measures the control that nurse managers have gained over budgeting. PMID- 10827688 TI - When releasing a temp nurse, give the agency good feedback. PMID- 10827689 TI - How to accomplish organization-wide competency and education. AB - To achieve high-quality, low-cost outcomes, all employees must be competent in their fields. Ensure one high standard for the entire organization with these seven steps to plan an organization-wide education and competency program. PMID- 10827690 TI - Give-and-take feedback. PMID- 10827691 TI - Wanted: behavioral skills. PMID- 10827692 TI - Orientation: six ways to avoid throwing new nurses to the wolves. PMID- 10827693 TI - Nurse executives champion change in integrated health systems. AB - When facilities join integrated health systems, nurse executives must increase their focus on governance, staff education, financial analysis, mentorship, and planning to maintain strong patient advocacy across a continuum of care. PMID- 10827695 TI - Take on the latest challenges for OR directors. AB - Operating room directors' responsibilities are changing and expanding with medical and technological advances, managed care, the nursing shortage, and market competition. PMID- 10827694 TI - Oh baby! A labor and delivery staffing system measures patient census and acuity. AB - The unique characteristics of labor and delivery units make it difficult to plan clinically and fiscally justified staffing patterns. This article presents an effective technique to measure patient census, track patient acuity, and determine required and actual nursing care hours. PMID- 10827696 TI - Purchasing influence. Top trends that support your stance. AB - Today's purchasing and supply professionals are changing the ways they work. Extend your nursing influence to the purchasing arena by learning the eight purchasing trends that will shape the industry. PMID- 10827697 TI - NIC puts nursing into words. AB - A standardized language promotes consistent thought and action--and raises a profession's profile. The Nursing Interventions Classification (NIC) arranges all of nursing's rich and varied interventions into a powerful, research-based tool. PMID- 10827698 TI - Critical care nurses read the writing on the wall. AB - Need a creative way to stimulate adult learners? Interactive bulletin boards book readers and give you an opportunity to communicate with humor. PMID- 10827699 TI - Communication systems make a caring connection. AB - Tips on how to select the nurse communication system most appropriate for your setting. PMID- 10827700 TI - Five deposition styles and how to handle them. PMID- 10827701 TI - Ask AONE's experts ... about a four-point plan to reduce medication errors. AB - Learn how you and your staff can implement a proactive four-point plan to reduce medication errors. PMID- 10827702 TI - Every country in the developed world has been reforming its health services. PMID- 10827703 TI - Let's hang on to what we've got.... PMID- 10827704 TI - Making up the numbers. PMID- 10827705 TI - Making an impact. PMID- 10827706 TI - Managed clinical networks: time to act. PMID- 10827707 TI - Nurse manager, change thyself. PMID- 10827708 TI - Dame Pauline Fielding. Interview by Tom Keighley. PMID- 10827709 TI - Many challenges ahead. PMID- 10827710 TI - Lessons from Scotland. PMID- 10827711 TI - Staying ahead of the competition. PMID- 10827712 TI - Nursing is a great profession! Spread the word. PMID- 10827713 TI - JCAHO solutions. Ins and outs of advance directives. AB - Nurses' responsibilities surrounding patients' advance directives can vary by setting. PMID- 10827714 TI - Protect your staff from workplace violence. AB - Your staff faces the possibility of workplace violence every day. Information and safety policies can give them a fighting chance. Answer the questions about workplace violence below, then check your answers at http:?www.nursingmanagement.com. PMID- 10827715 TI - How to establish a position and hours budget. AB - Learn how to convert your full-time equivalent (FTE) allocation into a position and hours budget. PMID- 10827716 TI - Integrated communication cascade: better information, faster response. AB - Learn how one hospital created an integrated communication cascade to increase efficiency and to maintain a consistent response to monitor-sensed rhythm changes. PMID- 10827717 TI - When an employee dies. AB - The death of an employee is a life-changing event. Learn how you and your employees can pull together to grieve, heal, and move on. PMID- 10827718 TI - Room at the top? AB - Nurse executives have joined hospital administrative teams, but are they accepted as fully integrated team executives? Learn how nurse executives and their influential colleagues view integration and its influences. PMID- 10827719 TI - The patient room of the future. AB - In the future, patient rooms will use high-tech to a human advantage. Learn how the new model provides a soothing, healing environment that increases nursing efficiency. PMID- 10827720 TI - Clinical service lines bring patients into focus. AB - In 1996, SSM St. Joseph Hospital of Kirkwood, Mo., was in crisis mode. A radical structure change from centralized nursing to clinical service lines helped to keep the organization in the black. PMID- 10827721 TI - Need critical care nurses? Inquire within. AB - A serious shortage of critical care nurses encouraged a nurse manager to develop intensive care unit (ICU) nurses through an innovative preceptor program. PMID- 10827722 TI - Have you and your staff signed self-care contracts? AB - Nurses are inevitable givers who ignore their own needs. Nurse leaders are in a key position to encourage staff members to care for themselves. PMID- 10827723 TI - Measure how ACNPs impact outcomes. AB - Nurse leaders need a way to measure acute care nurse practitioners' (ACNPs') performance and justify their positions. An objective documentation tool can help to evaluate ACNPs' work activities and measure performance and outcomes. PMID- 10827724 TI - Staffing and scheduling at your fingertips. AB - Learn how staff-scheduling software can help you reach goals for data gathering and management, outcomes improvement, or cost savings goals, and how to select the best system for your organization. PMID- 10827725 TI - Tips to help you recruit, manage, and keep Generation X employees. PMID- 10827726 TI - When and how to use adhesive film dressings. PMID- 10827727 TI - Using honey as a dressing for infected skin lesions. PMID- 10827728 TI - Guidelines for pressure ulcer prevention and management. PMID- 10827729 TI - Managing MRSA wound infection and colonisation. PMID- 10827730 TI - Managing leg ulcers: a review of the clinical guidelines. PMID- 10827731 TI - Taking a wound swab. PMID- 10827732 TI - When and how to use hydrocolloid dressings. PMID- 10827733 TI - Dog saliva complicates the healing of ulcers. PMID- 10827734 TI - Accurate documentation. PMID- 10827735 TI - Selecting compression hosiery. PMID- 10827736 TI - Managing burn blisters. PMID- 10827737 TI - MRSA 2000. PMID- 10827738 TI - The public health management of meningococcal disease. PMID- 10827739 TI - Body piercing: controlling the risk of infection. PMID- 10827740 TI - Which glove, when? PMID- 10827741 TI - [The Brocas report or how to connect the health care system and the paramedical professions]. PMID- 10827743 TI - [Care of the elderly: quality of life and autonomy]. PMID- 10827742 TI - [Nurses on humanitarian mission: towards the professionalization of volunteers]. PMID- 10827744 TI - [Care of the elderly: falls among the aged]. PMID- 10827745 TI - [Care of the elderly: devices and technical aids for the aged]. PMID- 10827746 TI - [Caring for the elderly: nutrition and the elderly]. PMID- 10827747 TI - [Care of the elderly: pain in the aged]. PMID- 10827748 TI - [Care of the elderly: local treatment of venous leg ulcers]. PMID- 10827749 TI - [Care of the elderly: anesthesia in the aged]. PMID- 10827750 TI - [Care of the elderly: Alzheimer's disease: the nurses are involved]. PMID- 10827751 TI - [Gastrointestinal disorders: 1. Functional digestive complaints]. PMID- 10827752 TI - [Management of sleep apnea syndrome]. PMID- 10827753 TI - [Disposal of hospital waste]. PMID- 10827754 TI - [The European nursing societies plead for a healthy Europe]. PMID- 10827755 TI - [Technical levels at the hospital] [In Process Citation] PMID- 10827756 TI - [Pharmaceutical satellite antennas: for a more efficient supplying of care units] [In Process Citation] PMID- 10827757 TI - [Sterilization: a priority unit within the pharmacy service] [In Process Citation] PMID- 10827758 TI - [The wandering computer at the hospital: medical record at the bedside] [In Process Citation] PMID- 10827759 TI - [Let us stop being sorry for ourselves. Interview by Anne Boiteux]. PMID- 10827760 TI - [The legacy of Desire Bourneville to the nursing profession]. PMID- 10827761 TI - [Patient education. Self supervision and self control of diabetes mellitus; the nurse's role. 2]. PMID- 10827762 TI - [Power and the nurse: from the obligation of healing to the power of thinking]. PMID- 10827763 TI - [Power and the nurse. The power of expression]. PMID- 10827764 TI - [Power and the nurse. A power without recognition]. PMID- 10827765 TI - [Power and the nurse. The power of sharing one's knowledge]. PMID- 10827766 TI - [Power and the nurse. Value and powers at the hospital]. PMID- 10827767 TI - [Power and the nurse. Power or nurses' power at the administrative council]. PMID- 10827768 TI - [Power and the nurse. Power, a tool for management]. PMID- 10827769 TI - [Power and the nurse. Let us find a unit with force and power]. PMID- 10827770 TI - [What to do in the presence of a generalized convulsive crisis in an adult]. PMID- 10827771 TI - [Proper usage of medicines. 6/12--aerosol therapy by nebulizers]. PMID- 10827772 TI - Parallel lists. PMID- 10827773 TI - Organisation of acute general hospital services: a surgical synopsis of the JCC document. AB - Hospital networks should be created, where possible by amalgamation into larger Trusts sufficient to provide comprehensive acute services to a population base of 450,000-500,000. In many areas, however, DGHs serving 200,000-300,000 will remain the backbone of the NHS and will provide the majority of acute services, although specific arrangements with neighbouring units will need to be made for a range of sub-specialties, such as vascular surgery. Where distance dictates the need for isolated small hospitals to continue to offer acute care, close professional links should be established with larger units so that patient care without compromise in quality can be offered even though transfer may be necessary. Because such hospitals may find it difficult to meet the new European Directive on the 48-hour working week for both permanent staff and junior doctors, staffing arrangements both at consultant and junior level might have to be very different from those expected in larger hospitals. There will need to be a major increase in the number of consultants in almost all surgical specialties. PMID- 10827774 TI - Help for basic surgical trainees: what the Raven Department of Education can offer. PMID- 10827775 TI - Chummy Sinnatamby--the man everybody knows. PMID- 10827776 TI - Perceptions of ENT training. PMID- 10827777 TI - Surgeons under stress (II). Update following the college seminar/workshop on pastoral care. PMID- 10827778 TI - Competencies required to be a competent surgeon. PMID- 10827780 TI - Re: 'Mr' or 'Dr'. PMID- 10827779 TI - Short listing for specialist registrars: consultants versus medical personnel. PMID- 10827781 TI - New distance learning courses from the Raven Department of Education. AB - The Raven Department of Education is marking the new century by developing two new distance learning courses, both of which will include computer-based learning using the College web site. STEP 2000 will be offered to basic surgical trainees from August 2000 and replaces the MRCS-STEP course for those preparing to take the MRCS examination. The second course is the department's first distance learning offer for higher surgical trainees. Designed for HSTs in general surgery in years 1-3, it provides an introduction to the surgery of breast disease. In the two articles below, the sessional tutors leading these developments outline the main features of the two courses and explain the educational principles behind them. PMID- 10827782 TI - Standardisation of the selection process for applicants for senior house officer posts for basic surgical training rotations. PMID- 10827783 TI - Commonwealth training initiative: step distance learning project. PMID- 10827784 TI - [Prescriptions of aerosol therapy. A survey of practices at the Besancon University Hospital]. AB - OBJECTIVE: Prescriptions of aerosol sprays concomitantly with other drugs can raise problems of incompatibility. METHODS: Medical practices in the clinical units of the Besancon University Hospital were analyzed to assess the therapeutic indications, the most frequently prescribed drugs, possible admixtures, the nature and volume of solvents used, drug protocols and type of aerosol therapy and nebulizer used. Sixty questionnaires were sent to all the units of the University Teaching Hospital of Besancon. RESULTS: Analysis of 48 questionnaires completed by head nurses showed that 28 different drugs and 26 different admixtures were prescribed. Only 2 of the admixtures had undergone prior validation. Only 7 (26%) of the drug formulations prescribed had received marketing approval. Recognized clinical practices for the administration of aerosol therapy were not applied and the aerosol sessions were not standardized. CONCLUSION: Many prescriptions are carried out without knowledge of the chemical compatibility of co-administered medicines. PMID- 10827785 TI - [Cost of medical imaging practices in acute abdominal syndromes]. AB - OBJECTIVES: To describe the costs of medical imaging practices in the diagnosis management of acute abdominal pain (AAP). METHODS: Medical imaging techniques until decision for treatment were prospectively recorded in patients presenting with AAP. Direct costs used hospital analytic accountability. Time of human resources involved was also surveyed prospectively. RESULTS: In 122 adult patients (2.3 examinations on average) before treatment decision making, the more frequent practices were: initial plain abdomen x-ray followed by tomodensitometry (36.8%), by echography or endoscopy (17.2%), plain abdomen solely (19.6%) or initial abdominal tomodensitometry (12.3%). Direct costs ranged from 977 to 1073 FF for practices with initial plain abdomen x-ray, and from 996 to 1150 FF with initial tomodensitometry. It ranged from 808 to 880 FF when the treatment decision was surgery, and 300 FF higher when it was medical. CONCLUSION: Differences in costs assessed for practices were very narrow. Such information should be taken into account to determine cost-effective strategies, and to built up reference guidelines. PMID- 10827786 TI - [Myasthenia in the aged: a case with unusually late onset]. AB - BACKGROUND: Myasthenia is an uncommon autoimmune condition that can occur at any age. Peak frequency is seen around the age of 65 years. We report a case with a particularly late onset and discuss the particular conditions of myasthenia in the elderly subject. CASE REPORT: A 97-year-old patient was hospitalized for dysphonia and dysphagia associated with exercise-induced dyspnea. The general picture suggested generalized myasthenia confirmed by the electromyography exploration and a positive anticholinesterase test. Treatment with acetylcholinesterase inhibitor was effective although cure was incomplete. Further improvement was obtained with immunosuppressor therapy using azathioprine. DISCUSSION: The clinical presentation of very late onset myasthenia differs little from that in younger subjects excepting the very high frequency of brain stem involvement in the initial presentation. Diagnosis may however be more difficult as other conditions are more easily taken to be the causal element. Thus, for the elderly patient, the real problem is to envisage the diagnosis of myasthenia. Positive diagnosis is based on the same criteria as in younger subjects. Clinicians should be aware of the possibility of myasthenia in the geriatric population as specific treatment can improve functional prognosis with satisfactory efficacy. PMID- 10827787 TI - [Pseudotumorous cystic lesion of the thigh caused by Echinococcus granulosus]. PMID- 10827788 TI - [Rheumatic manifestations in Buerger's disease]. PMID- 10827790 TI - [Therapeutic efficacy of bed rest should be reevaluated]. PMID- 10827789 TI - [Septic arthritis caused by Aeromonas hydrophila]. PMID- 10827791 TI - [Cyclotrons in cancerology: proton therapy used in uveal melanoma. Synthesis of clinical evaluation and guidelines]. PMID- 10827792 TI - [Cannabis-induced arteriopathy]. PMID- 10827793 TI - [Diagnostic value of antiphospholipid antibody assays]. PMID- 10827794 TI - [Effects of cognitive aging on memory performances]. AB - PSYCHOLOGY OF AGING: This article sums up studies on influence of normal aging in memory. Aging is a process involving many dimensions: Psychological, Biological and Social. These elements interact with each other and needed to be considered in the description of human memory aging. COGNITIVE APPROACH OF HUMAN MEMORY: Moreover, researches on human memory have been conducted within the framework of current theoretical views of memory. The present review provides an outline of the cognitive neuropsychology models of memory, its nature, components and organization. CLINICAL APPROACH OF AGING AND MEMORY, CONSEQUENCES: This introduction may help the reader to understand more clearly empirical evidence of age-related differences in memory and attentional functioning. In closing, the human factors psychologists must be take in account while adopting a global approach of the understanding of memory aging. Psychological field provides a complete structure for assessing human being and is helpful before to conclude between normal or pathological memory aging. PMID- 10827795 TI - [Memory complaints: epidemiology and diagnostic approach]. AB - IN THE ELDERLY: Complaints of poor memory for everyday activities are common in the young as well as in the elderly. However, in the elderly, memory complaints are especially of interest on account of their frequency and the fear of being related to early Alzheimer's disease. Two types of memory complaints should distinguished. ENCODING/STORAGE DEFECTS: Memory difficulties related to early Alzheimer's disease are mainly related to a deficit in encoding and/or storage. They are restricted to the recent past and rapidly associated with temporal disorientation, limitation in everyday activities and behavioral modifications alerting the family responsible for seeking medical advice in most cases. Short examination of memory performance shows a free recall deficit which is not improved by cued recall. BENIGN DIFFICULTIES: Memory difficulties of benign type are due to a pure recall deficit: they are experienced as well for ancient as for recent past and the difficulty in recall is transient. There are no behavioral modifications or limitation in everyday activities: the disorder is purely subjective. At examination, the deficit in free recall is corrected by cueing. Benign type of memory complaints is mostly related to psychoaffective disturbances and subjects experiencing this type of memory difficulties are not at high risk for developing dementia. PMID- 10827796 TI - [Memory clinics: value and limitations]. AB - CONCEPT: Memory clinics have developed since the early 1980s. Their main objective is the diagnosis of dementia for early multiple disciplinary management. INTEREST: Memory clinics serve an expert center, allowing early diagnosis in the predementia phase and a precise diagnosis of the cause. Memory clinics have revealed the frequency of psychiatric disorders, particularly depression and anxiety, leading to memory complaints. They provide a significant contribution for follow-up of dementia patients, announcement of the diagnosis, initiation of new treatments, prevention of "emergency hospitalizations" as well as postponement or preparation of institutionalization. They also have an educative role, promoting better use of psychotropics and a better understanding of management approaches and incidents observed during the disease course. Clinical research, which is not always sufficiently supported, is one of their important responsibilities. LIMITATIONS: The basic limitation for memory centers is the problem of the availability of qualified personnel. The lack of sufficient interest in truly early screening for Alzheimer's disease remains a problem to be solved. PMID- 10827797 TI - [Alzheimer type dementia: is early diagnosis significant?]. AB - ADVANTAGES OF EARLY DIAGNOSIS: Better prognosis and quality of life for patients with Alzheimer's disease and their caregivers depends on early diagnosis as specific treatment (anticholinesterase drugs) early in the disease process can have a beneficial effect on cognition and psychiatric or behavioral disorders. In addition early diagnosis gives the physician the opportunity to provide adapted advice for the patient and caregivers especially important in preventing complications and helping the family cope with the inevitable disruption of the family pattern caused by the disease, a situation which is particularly for the "designated caregiver". PREDEMENTIA STATES: The question of early diagnosis raises several types of problems. Defining the limits of the disease is particularly difficult: when does Alzheimer type dementia start? what is the definition of predementia? A growing body of work suggests that it is warranted to identify patients at risk of developing Alzheimer type dementia since, according to certain authors, they can benefit from specific treatment. RISK FACTORS: The only fully recognized risk factors are age, family history of dementia and presence of the allele epsilon 4 of the apolipoprotein E gene. There are probably several other risk factors. Their identification is a current subject of debate. TOOLS FOR EARLY DIAGNOSIS: Psychometric tests have been shown to provide specific information useful for interpreting the clinical assessment which must focus on detecting early signs and exploring even minimal memory deficiencies. PMID- 10827798 TI - [Endonasal dacryocystorhinostomy: a personal experience]. AB - Stenosis of the naso-lachrymal duct as a consequence of acute or chronic inflammation, trauma or congenital malformation is a relatively frequent lachrymal duct pathology. The symptoms include epiphora and intermittent tumefaction of the medial ocular canthus, indicative of a deficiency in lachrymal drainage. If such conditions do not respond to medical treatment, lavage and transcanicular probe, surgery proves necessary. Modern techniques for endonasal surgery place the otologist in the forefront in the surgical treatment of this disorder. Endonasal dacryocystorhinostomy (DCR) is indeed a valid alternative to traditional extra-nasal surgery; once exclusively the domain of the ophthalmologist. However, it does require a certain expertise in micro-endoscopic surgery. In most cases sophisticated equipment is not required and the procedure can be performed with just a few instruments: electric knives, scalpel and Citelli forceps. Endonasal DCR makes skin incisions unnecessary and makes it possible to reveal, and possibly, correct, any sequela subsequent to traditional DCR such as ethmoidal rhino-sinusitis or the presence of adhesions. In the present work 67 patients--24 with recurrence after traditional surgery--underwent endonasal DCR within the last 22 months and with an average follow-up of one year. Surgery was successful in 90% of the cases. PMID- 10827799 TI - [The accuracy of the fine needle aspiration biopsy in 1250 thyroid nodules]. AB - Between 1989 and 1998 a total of 1250 thyroid nodes underwent Fine Needle Aspiration Biopsy (FNAB). Of these 150 went on to surgery with subsequent histological examination which proved positive for malignant neoplasm in 35 nodes. The remaining 115 nodes presented benign lesions. The cytological diagnoses were preventatively broken down into three groups: a) benign; b) malignant; c) suspected malignancy. Group a) included 53 nodes; histology confirmed the diagnosis of a benign lesion in 50 of these nodes (True Negatives), while 3 proved malignant (False Negatives). The cytological diagnosis of malignancy was reached in 24 nodes (group b) and subsequent histology confirmed the malignancy in 18 cases (True Positives) while the remaining 6 nodes tested negative for neoplasm (False Positives). Group c) included those thyroid nodes which cytology classified as follicular neoplasms and for which histology was required to reach a diagnosis of malignancy or benignity; for this reason these cases were not used in the evaluation of diagnostic reliability. Of these 15 (20.5%) proved malignant and 58 (79.5%) benign (44 follicular adenomas and 14 micro-macrofollicular struma nodes). On the basis of the above data, the diagnostic accuracy of FNAB is 88.3%, sensitivity 85.7% and specificity 89.3%. These findings are substantially in agreement with the international literature which considers cytological testing highly reliable. In analyzing the cases which were not confirmed by histology, it was interesting to note that among the 4 false positives--defined as the "presence of atypical cells in a lymphocyte infiltration context"--a full three were thyroadenitis nodes for which the presence of atypical cells is quite common. The three false negative nodes, on the other hand, included two cysts for which cytology did not reveal neoplastic cells. The present experience suggests the following: 1) FNAB is still the most reliable technique for the diagnosis of thyroid neoplasms; 2) the presence of atypical cells in thyroid node lesions is not always indication of a malignancy; 3) cysts must be subject to careful follow-up since they can mask a malignant neoplasm. PMID- 10827800 TI - [Tc99m-sestamibi SPECT and head and neck tumors: preliminary data]. AB - After underlining recent progress in nuclear medicine diagnostics, even in oncology, the authors describe the most important methods used and oncotropic capacity of some radiodrugs currently used in clinical practice: i.e. perfusion tracers. Then they present the results of a personal experience using Tc99m SestaMIBI SPET in detail. Fifteen patients suffering from various primary head and neck neoplasms underwent systematic clinical evaluation, endoscopy, CT and/or MRI and Tc99m-SestaMIBI SPET. The authors then briefly describe the method and underline the fact that the literature lacks any homogeneous data on the use of Tc99m-SestaMIBI SPET in the study of head and neck neoplasms. The results indicate that the method is reliable in identifying both primary tumors and laterocervical metastases. Then, taking indication from the only case currently available on post-radiotherapy recurrence, the authors suggest an interesting field of application for this technique: the study of post-radio-chemotherapy and/or surgery recurrences. In conclusion, the authors underline the valid contribution this method currently offers both in diagnosis and follow-up as well as its future of ever expanding clinical applications. PMID- 10827801 TI - [Acute epiglottitis in children: current criteria for the diagnosis and treatment]. AB - Acute epiglottitis is still a potentially lethal pathology, particularly in early childhood. The present study involves seven cases of acute epiglottitis in children under 4 years of age. The authors describe the diagnostic and therapeutic protocols used in these pediatric patients placing particular emphasis on the use of endoscopy and the need for prompt hospitalization in an intensive care unit to best integrate the diagnostic approach with therapeutic treatment. PMID- 10827802 TI - [Tumors of the parapharyngeal space: case report of clear cell myoepithelioma of the parotid gland and review of literature]. AB - The parapharyngeal space is a deep neck region and can be the site of a wide range of neoplasms, including primary benign or malignant tumors, metastatic tumors, neoplasms invading the parapharyngeal space from the adjacent regions and neoformations stemming from systemic processes. Tumors of the parapharyngeal space are, nevertheless, relatively rare and account for 0.5-0.8% of all head and neck tumors. Most are benign tumors originating in the deep lobe of the parotid gland and the minor salivary glands. The most frequent tumor is pleomorphic adenoma while myoepithelioma accounts for just 1% of all salivary gland tumors. The present case report involves a case of myoepithelioma of the deep parotid gland lobe, arising in the parapharyngeal space. Together with a review of the literature, considerations are presented on the clinical and radiological evaluation, differential diagnosis and treatment of these lesions. PMID- 10827803 TI - [Maxillary ameloblastoma: a case report]. AB - Ameloblastoma is a neoplasm of odontogenic origin. Although it is considered a benign lesion it presents some peculiarities including a high local recurrence rate, particularly after conservative surgery, and a high loco-regional invasiveness. The present work describes a case of ameloblastoma of the left maxillary sinus bordering on the homolateral nasal fossa. The male patient was admitted to the E.N.T. Dept II of the University of Turin in May 1998 and underwent left radical maxillectomy. The authors also review the literature on the topic. PMID- 10827804 TI - Different types of interferon for the therapy of HCV chronic active hepatitis in the elderly patient. AB - HCV correlated hepatitis is a pathology on the increase, and it is especially affecting patients above 60 years old. The only treatment for this disease is therapy with different types of interferon. The authors take into examination three of their previous studies on treatment of HCV correlated chronic hepatitis in the elderly using different types of interferon: recombinant interferon alpha, interferon beta, and lymphoblastoid interferon, in order to evaluate which one, among the three, should be the best for the treatment of this pathology in the elderly. The data show that recombinant interferon alpha is preferable since the remission percentage is higher (75%), compared to beta (53.8%) and lymphoblastoid interferon (60%). As far as the relevant side-effects in elderly patients are concerned, beta-interferon therapy is almost with no side-effects. Even in cases where there could be a possible higher exposure to side-effects linked to the use of recombinant interferon alpha, still, the risk/benefit ratio suggests that this particular drug should be used for treating this pathology in elderly patients. PMID- 10827805 TI - Adrenomedullin assay and its clinical significance. AB - Adrenomedullin (Am) is a recently discovered peptide, first purified from pheochromocytoma specimens, with a chemical structure similar to that of CGRP and amylin. Adrenomedullin is present in numerous human body tissues and its powerful vasodilatatory activity is thought to play an essential role in cardiovascular and renal homeostasis. PMID- 10827806 TI - Lithium safety in the prophylaxis of bipolar disorders: a study with plasma levels. AB - From many decades efficacy of lithium salts, as mood stabilizers, has been largely recognized, but their tolerability, in particular during intermediate or long-term treatments is still discussed. The most frequently described side effects can affect several organs. Aim of the study was to evaluate lithium carbonate tolerability after a "brief" (1 month-4 years), "intermediate" (5-9 years) and "longterm" (10-21 years) treatment of patients affected by Bipolar Disorders (BD). 27 patients (14 males, 13 females), aged from 20 to 78 years (mean 49.03 years +/- 14.61 SD), affected by BD, type I, according to DSM IV criteria were included into the study. Our data suggest a good tolerability of lithium salts without significant differences among the three different periods of treatment. PMID- 10827807 TI - Migraine with visual aura in developing age: visual disorders. AB - Visual disorders are an important symptom in the migraine of developing age. Different kinds of visual disturbances can precede, accompany or follow a migraine attack. These visual disturbances can be grouped into negative (hemianopsia, quadrantopsia, scotoma) and positive (phosphene, teicopsia, metamorphopsia, macropsia, micropsia, teleopsia, diplopia, dischromatopsia, hallucination disturbances) disorders. The pathogenetic mechanism of the visual phenomena of migraine has not yet been clarified. Various hypotheses have been proposed: vasospasm with consequent ischemia of some cerebral areas, the opening of arteriovenous shunts between the intra and extra cerebral circulation, the formation of microthrombi in arterioles and dopaminergic hypersensitivity of some nervous centers. We have studied 1787 children, affected by migraine with (13%) or without (87%) aura. Among the patients, 211 (12%) referred visual disorders, especially scotoma and phosphene. These data let us hypothesize that a relationship between migraine and visual disorders is present also in pediatric age. However this relationship is less important than in adults. PMID- 10827808 TI - Acute otitis media. Histopathological changes: a post mortem study on temporal bones. AB - A classification of otitis media is proposed and the histopathological changes of otitis media with effusion are reviewed post mortem on two temporal bones. A case of adhesive otitis media is observed on temporal bone slides. The clinical and therapeutic aspect are examined and treatment guidelines are proposed. PMID- 10827809 TI - High resolution computed tomography (HRCT) assessment of beta 2-agonist induced bronchodilation in chronic obstructive pulmonary disease patients. AB - The evaluation of airway obstruction reversibility in chronic obstructive pulmonary disease (COPD) patients is currently performed by means of an indirect assessment of drug inducing variations in pulmonary function tests. Imaging techniques, especially high resolution computed tomography (HRCT), usually provide a complete evaluation of lung parenchyma (bronchial and vascular structures), but so far they have never been applied to visualize the effects on the bronchopulmonary tree of some pharmacologic stimuli (beta 2 adrenergic agonist), currently used in clinical practice to disclose the presence of airway reversibility. In order to assess the possible role of HRCT in this setting, five COPD patients have been subjected to a double functional radiologic evaluation before and after salbutamol-induced broncho-dilation, with a rigorous assessment of bronchial diameter changes by means of "bronchus-vessel" ratio, currently used for diagnosing bronchiectasis in COPD patients. The results of this experimental study enable us to visualize drug induced broncho-dilation, with a direct assessment of airway reversibility in these patients, and to show a good correlation between functional and HRCT findings, raising the possibility of evaluation COPD patients with abnormal spirometric results or with early signs of lung involvement. PMID- 10827810 TI - Electroionotherapy in acute arthrorheumatic pain. AB - The authors evaluated the efficacy of an electronic treatment for pain, using an ion flow generator, BE-101 model by Bio-Ejt, on 19 patients suffering from acute pain of an arthrorheumatic nature. Each patient was treated for two weeks every other day (6 sittings), each sitting lasting 20 minutes at an intensity of about 30 microAmper for both transducers. The results demonstrated that this technique is very effective in curing the pain. PMID- 10827811 TI - Specific immunotherapy in children: past and present. AB - Specific immunotherapy (SIT) in children is usually completely neglected. Pediatric SIT being not an optional treatment should be administered as soon as possible, also in children aged 2-3 years, due to the very early asthma and rhinitis onset, contrarily to a recent publication that continues to stress the danger of anaphylactic reactions without displaying reliable data. On the other hand drugs represent only a symptomatic treatment, therefore SIT is the only treatment which can alter the natural course of respiratory diseases. PMID- 10827812 TI - A randomized openly comparative study between rifaximin suspension versus rifaximin pills for the eradication of Helicobacter pylori. AB - BACKGROUND: It has been recently shown that Rifaximin, although given as a suspension, plus omeprazole, may be a promising antibiotic against Helicobacter pylori (H pylori) and worthy of further study. AIM: We have therefore evaluated Rifaximin suspension versus Rifaximin pills, in a randomly openly allocated fashion study, in H pylori positive patients. METHODS: Twenty patients with upper gastrointestinal symptoms (M/F: 13/7, age range 28-68; mean 49.6 yrs) were found to have H. pylori associated gastritis. They were allocated in an open randomized study to two different treatment groups for two weeks: (A) Rifaximin suspension 1800 mg three times a day plus Omeprazole 20 mg twice a day (n = 10), (B): Rifaximin pills 1800 mg three times a day plus Omeprazole twice a day (n = 10). Symptoms such as pirosis, bloating, epigastric pain and nausea were recorded by diary card and were evaluated before and four weeks after stopping treatment. Patients were assessed by endoscopy, histology and urease testing at entry and four weeks after stopping treatments. All the twenty patients were available four weeks after stopping treatment. RESULTS: A statistically significant improvement of the symptoms were found overall after Rifaximin treatments for pirosis, bloating, epigastric pain (p < 0.001 respectively). A significant difference in the symptom's score at the end of the two treatments were recorded between the two groups for bloating alone (p < 0.070). A different and major fall in the neutrophils, between the two treatments was observed with Rifaximin pills compared to Rifaximin suspension. The same observation was obtained according to the intensity of H. pylori reaching an eradication rate of 40% and 60% for Rifaximin suspension versus Rifaximin pills plus omeprazole respectively. In conclusion, these data suggest that Rifaximin pills may be an effective antibiotic against H pylori and worthy of further study. PMID- 10827813 TI - The infrequent association of synchronous renal and colonic malignancies. AB - The coexistence of both kidney and colon primary malignancies is a rare condition. We report the case of a 75-year-old woman who presented with bilateral pulmonary nodules at chest X-ray and stratigraphy. Total-body CT scan showed multiple, apparently metastatic, bilateral pulmonary lesions, a diffusely dysomogeneous neoformation in the lower pole of the right kidney and a gross neoformation in the ascending colon. A right nephrectomy and a right hemicolectomy were performed and histology showed two primary neoplasms: clear cell renal carcinoma and undifferentiated adenocarcinoma of the colon. PMID- 10827814 TI - Metastasis to the breast of a renal carcinoma: a clinical case. AB - The breast is an unusual site for metastatic disease, and generally the lesion appears same times after the diagnosis of the primary malignancy. Authors report a case of a breast metastasis from a hypernephroma, six years after nefrectomy. PMID- 10827815 TI - Pharmacokinetics of sebacic acid in rats. AB - The pharmacokinetics of disodium sebacate (Sb) was studied in Wistar rats of both sexes. Sebacate was administered either as intra-peritoneal (i.p.) bolus (six doses ranging from 10 mg to 320 mg) or as oral bolus (two doses: 80 and 160 mg). Plasma and urinary concentrations of Sb and urinary concentrations of Sb and its products of beta-oxidation (suberic and adipic acids) were measured by an improved method using gas-liquid chromatography/mass-spectrometry. A single compartment with two linear elimination routes was selected after no increase in significance was shown by an additional compartment and after a saturable mechanism was found to be unsuitable. Both renal and non-renal elimination parameters were obtained by Marquardt non linear fitting of plasma concentrations together with urinary elimination. The data reported are calculated from the analysis on the whole population of rats and referred to an average body weight (bw) of 100 g. The Sb half-time was 31.5 min. The tissue elimination rate was 0.0122 min-1. The overall volume of distribution was found to be 26.817 ml/100 g bw. The renal clearance was 0.291 ml/min/100 g of bw, which is much less than the value of GFR reported in literature (about 1 ml/min/100 g bw), suggesting the presence of Sb reabsorption from the ultrafiltrate. The value of Sb renal clearance was found to be a concentration-independent function, suggesting the presence of a passive back-diffusion. The relative bioavailability of the oral form compared to the i.p. form was 69.09%, showing a good absorption of the drug. PMID- 10827816 TI - Meniere's disease: is it a bilateral disease? AB - BACKGROUND: Bilateral Meniere's disease (MD) is still controversial due to different criteria used to assess the involvement of the primarily affected ear and the contralateral one. We evaluated the percentage of bilateral forms in 49 patients with MD. METHODS: 49 patients with (MD) were studied. All were selected according to the following requirements: history, tonal audiometry, glycerol test, Auditory Brainstem Response (ABR), vestibular examination. Magnetic Resonance (MR) was performed in 14 patients. RESULTS: A raised hearing threshold in the contralateral ear was found in 23 patients, but only 7 (14.3%) fulfilled the requirements to be considered affected by bilateral MD. The delay of occurrence in the contralateral ear was 7 years (from 5 to 12 years). Submillimeter Magnetic Resonance is determinant for differential diagnosis with Meniere-like syndromes. CONCLUSIONS: A conservative approach in surgical treatment of unilateral Meniere's disease is recommended because of the possibility of evolution in a bilateral form, that can occur even after 10 years from the onset of the disease. PMID- 10827817 TI - Cholesterol granuloma of the petrous apex. A post mortem study on temporal bones. AB - Otitis media (OM) is an infection localized in the middle ear: mastoid, middle ear cavity, Eustachian tube. The classification of OM includes otitis media with effusion, otitis media without effusion, and chronic otitis media. A rare complication of chronic otitis is cholesterol granuloma of the petrous apex. It may develop in any aerated portion of the temporal bone but most commonly develops when a pathologic process obstruct the air cell tracts to the petrous apex preventing normal aeration. PMID- 10827819 TI - Feeding high-risk infants with family history of allergy. AB - Since the turn of the century, CM (cow's milk) formulas have become progressively more common as breast milk substitutes when mother's milk is unavailable, and CM allergy (CMA) has thus gradually become a more common disorder. Therefore the management of CMA infants confronts pediatricians and allergists with one of the most demanding challenges. In the first year of life of many children, CM provides almost the entire dietary supply of proteins, carbohydrates, and fat; its high nutritional value and low cost should be noted. However, children with CMA can avoid CM without nutritional loss if nutrients are provided by other foods such as meat, fish, vegetables and fruit. Thus the choice of an adequate CM substitute for high-risk infants with family history of allergy is mandatory. The ideal CM substitute should be hypoallergenic; have an adequate nutritional value according to the infant's age; be easily available and inexpensive; and be palatable in order to obtain a good compliance. CM substitutes now available are soy protein formulas (SPF) and hydrolysate formulas (HF). Consequently pediatricians are now over-whelmed with a variety of information on new formulas called hypoallergenic and are faced with a difficult choice among them. PMID- 10827818 TI - CGRP and ET-1 plasma levels in normal subjects. AB - OBJECTIVE: Calcitonin gene-related peptide (CGRP) is a 37 amino acid peptide displaying about 50% homology with amylin which is secreted from the pancreatic islets of Langerhans. The main form, the beta-CGRP, is produced by the enteric nervous system and perivascular nerves of the vasa-vasorum. It represents one of the most powerful vasodilator yet discovered but its role is not yet completely clarified. High levels of this peptide have been shown in patients affected with thyroid medullary carcinoma, phaemocromocytoma and lung carcinoma. Recently circulating levels of CGRP have been found in normal subjects. Endothelin-1 (ET 1), a potent vasoconstrictor peptide, isolated from porcine endothelial cells, is an important regulator of the vascular tone acting in physiological antagonism with atrial natriuretic hormone (ANH). With this study we intended to investigate the presence of any correlation between CGRP and ET-1 in normal subjects. PATIENTS: For the study we considered 20 normal subjects (11 males and 9 females) aged 23 to 50. MEASURES: Plasma levels of CGRP and ET-1 were measured by radioimmunological Kit. RESULTS: A positive and significant correlation between calcitonin gene-related peptide and endothelin-1 was found. CONCLUSIONS: Our results confirms that CGRP and ET-1 have opposing actions on vessels and that they can act together in haemodinamic regulation. PMID- 10827843 TI - [Delirium? Admission to the general hospital, rather than a psychiatric hospital]. AB - Delirium is often not recognized either by psychiatrists or by other physicians. In two men, aged 78 and 70 years, a diagnosis of delirium was not recognized at admission. The first patient had Alzheimer's disease with superimposed delirium caused by pulmonary embolism, which led to cardiopulmonary arrest during his stay in a psychiatric hospital. The other patient had a delirium due to multiple somatic causes (carbon monoxide poisoning with cerebral damage, pulmonary infection, heart attack) which was not recognized leading to an early discharge from the general hospital. The first patient died later on because of complications of a pneumonia and the other patient was transferred to a nursing home on a maintenance dose of haloperidol. Since delirium is a symptom of a medical disorder, delirious patients should be referred primarily to a general hospital. PMID- 10827844 TI - [Lavage almost never indicated after an autointoxication]. AB - In many Dutch hospitals gastric lavage leaving charcoal and a laxative is the treatment of choice after autointoxication. Gastric lavage is not without risks. No difference has been demonstrated in efficacy and safety of gastric lavage combined with administration of activated charcoal on the one hand and just administration of activated charcoal on the other. In theory, gastric lavage might be useful in patients presenting shortly after the selfpoisoning (up to 2 hours) and in the case of delayed gastric emptying up to 4 hours. Gastric lavage is indicated without doubt in patients presenting shortly after ingestion of toxic substances which are poorly adsorbed by charcoal (for instance lithium). In case of a mild intoxication (for example with benzodiazepines), the risks of drug toxicity do not outweigh those of lavage, regardless of the time elapsed after ingestion. If gastric lavage is applied, it should be performed properly, i.e. with warm water (38 degrees C), with a 36-40 Fr. tube, using aliquots of 200-300 ml. In a minority of the intoxications whole bowel lavage should be employed. PMID- 10827845 TI - [Use of margarine fortified with phytosterols as a therapeutic food]. AB - The intake of one tub of margarine with added phytosterols per week has been shown to reduce plasma total and low density lipoprotein (LDL) cholesterol levels by 9 and 13% respectively, with no effect on high density lipoprotein (HDL) cholesterol or triglyceride levels. This reduction in cholesterol concentration may help to decrease the risk of coronary heart disease by 20 to 50%. No significant side effects have been reported. Phytosterols reduce cholesterol levels by inhibition of cholesterol absorption in the intestine. The margarines with added phytosterols are an example of functional foods (novel foods) which are designed to enhance health or contribute to reduction in a specific disease risk. In 1997, the European Union has introduced rules for the evaluation of novel foods, the Novel Food Regulatory Approval Procedure. Post-marketing surveillance of functional foods is obligatory to control long term safety and efficacy. PMID- 10827846 TI - [Cloning: applications in humans. I. Technical aspects]. AB - The successful cloning experiments in mammals such as the sheep and mouse prompted speculations on clinical application in humans. Cloning is possible by nucleus transplantation and by embryo splitting. Nucleus transplantation does not result in a genetically completely identical individual because the mitochondrial DNA originates from the ovum donor. Embryo splitting may be regarded as the artificial production of a monozygotic multiplet. Possible applications of cloning in humans belong in the context of reproduction (treatment of couples with subfertility, with genetic problems or with a 'replica motive'), transplantation of genetically identical tissue, and scientific research. PMID- 10827847 TI - [Cloning: applications in humans. II. Ethical considerations]. AB - Reproductive cloning in adults/children evokes unfavourable reactions. Direct objections are that cloning is unnatural, that it affects human dignity and violates the individual's right to genetic uniqueness. Consequential objections concern unjustified health risks for the progeny, unjustified psychosocial risks for the clone child and the risk of cloning for eugenetic purposes. There is consensus that reproductive cloning of existing persons is unjustifiable as yet because of the health risks for the offspring. Reproductive cloning of embryos is possible by means of nucleus transplantation and of embryo splitting. The ethical analysis of reproductive cloning of embryos depends on the purposes and applications. At least some of the moral objections against cloning of adults/children are not or not completely applicable to reproductive cloning of embryos. Conditions to be put to reproductive cloning of embryos are efficacy, safety and, at least for the time being, avoidance of asynchrony in transferring identical embryos. The ethical aspects of its application in the context of genetical reproductive techniques must be evaluated separately. Therapeutic cloning may be acceptable if alternatives are lacking. PMID- 10827848 TI - [Vaccination against acute otitis media]. AB - Acute otitis media (AOM) is the most frequent bacterial infection in childhood. Because of the high morbidity, the costs of AOM and growing concern about increasing resistance of pneumococci, the most common bacterial cause of AOM, prevention of AOM is important. Vaccination with the recently developed pneumococcal conjugate vaccines leads to a reduction in the number of AOM cases caused by the serotypes present in the vaccine, but the reduction in overall AOM incidence is below 10%. In particular the children with recurrent episodes of AOM may benefit more from these pneumococcal conjugate vaccines. PMID- 10827849 TI - [Identification of a new mutation (CysII6Gly) in a family with neurogenic diabetes insipidus]. AB - OBJECTIVE: Elucidation and identification of the molecular biological alteration in the arginine-vasopressin neurophysin II AVP-NPII gene in a family with familial neurohypophysial diabetes insipidus (FNDI). DESIGN: Descriptive. METHODS: Following the finding of diabetes insipidus in a 2-year-old boy and his father a molecular genetic investigation was performed in the Isala Klinieken, Sophia location, in Zwolle, the Netherlands, to determine the nature of a possible gene mutation. Thereafter the AVP-NPII gene was screened in the family with polymerase chain reaction (PCR) and restriction enzyme analysis on DNA isolated from peripheral blood. An extensive pedigree was made. RESULTS: A new mutation in the AVP-NPII gene was identified in the part encoding the transport peptide neurophysin II. in exon 3, on codon 116, in which thymine was replaced by guanine, leading to the amino acid glycine instead of cysteine in the gene product. CONCLUSION: In a Dutch family with familial neurohypophysial diabetes insipidus a new gene mutation was found (CysII6Gly). Clarification of the molecular background of FNDI in this family made it possible to test family members in a relatively simple and friendly way (without the thirsting test) by PCR and restriction enzyme analysis for the presence of the mutation and the predisposition for diabetes insipidus. PMID- 10827850 TI - [Diverticulosis of the small bowel]. AB - A 77-year-old man known with chronic abdominal complaints and a malabsorption syndrome presented with ileus, peritonitis and air in the abdominal cavity. Surgery showed perforation of a jejunal diverticulum and extensive small bowel diverticulosis. The pathologic segment was surgically removed but the patient died afterwards of septic shock, diffuse intravascular coagulation and multi organ failure. Small bowel diverticulosis has a prevalence of 0.3-2.5% and symptoms are present only in the minority of the cases. Apart from bacterial overgrowth and malabsorption, complications include bleeding, mechanical obstruction, volvulus and perforation. Diagnosis is difficult and mostly made by double-contrast radiology of the small bowel or during laparotomy. Treatment can be conservative in most patients; only in case of severe and persistent symptoms or complications like ileus, bleeding, perforation or volvulus, resection of the affected bowel segment is indicated. PMID- 10827851 TI - [Medicine inspired by poverty]. AB - Since his arrival in Egypt in 1994 the author joined a number of archaeological expeditions as a surveyor and part-time physician. During this latter activity he came into contact with the beliefs and practices of the local workmen and those of the Ababda Bedouin in particular. Living a harsh life in the southern part of the Egyptian Eastern Desert, their medicine seems to be inspired by poverty. Widely used for all internal disorders are 'kaya bil-naar': oval scars made with red-hot metal instruments. Another, less common, form of scarification is 'mi'ah hed'asher', three parallel lines carved deeply into the cheek of the patient. 'Muhawy' is the bite of a snake into the earlobe of the patient, to prevent snake bites. Another prophylactic is the 'higab', a small leather pouch containing a magical object or text. Therapies for less serious disorders include the use of herbs, spices and foodstuffs, often prepared in special ways. Externally, car fuel and axle grease are widely used. With the development of the Red Sea coast for tourism, the life of the Ababda Bedouin will change fundamentally. The above practices are likely to be replaced by Western medicine, probably a change for the better for these people. PMID- 10827852 TI - [Diagnosis and treatment of cholelithiasis]. PMID- 10827853 TI - Plans to hasten death among gay men with HIV/AIDS: relationship to psychological adjustment. AB - This study assessed the prevalence and nature of thoughts and future plans to end one's life in a group of gay men with HIV/AIDS over an 18-month period. HIV positive men (n = 167) participated in a series of clinical interviews which measured current health status, current and past psychiatric disorders, current levels of distress, and thoughts and plans about ending their lives currently or at some future point. A small number of HIV-positive men (17%) reported serious thoughts or plans to end their lives at some point in the future which were stable over an 18-month period. No differences in psychiatric or medical measures were observed among men with and without thoughts/plans at either assessment point. In the absence of current psychiatric disorders, such thoughts or plans about the future may represent one way to maintain control and independence in the face of the uncertainty of life with HIV illness. PMID- 10827854 TI - Training of obstetric nurses and HIV testing in pregnancy at a managed care organization. AB - The purpose of this study was to evaluate the effectiveness of an HIV Training Programme for obstetric nurses at a managed care organization. Nurses were trained to counsel pregnant women about the HIV antibody test according to the latest recommendations made by the US Public Health Service and the Texas legislature. Training improved the nurses' overall knowledge about the content of the programme and nurses were more likely to offer the HIV test to all pregnant women. Still, contrary to what was predicted, the nurses did not give more information to increase the knowledge pregnant women had about HIV infection, transmission and available treatments. However, pregnant women who received prenatal care after the nurses attended the programme were more likely to be tested than women who received prenatal care before the nurses were trained (96% versus 48%); in turn, the likelihood that high-risk women would be tested for HIV also increased (94% versus 60%). There was no evidence that mandatory testing with right of refusal would deter pregnant women from being tested for HIV. The increased rate of HIV testing was attributable to the providers asking their patients to be tested rather than on their patients' knowledge of its benefits. PMID- 10827855 TI - Work-related stress and occupational burnout in AIDS caregivers: test of a coping model with nurses providing AIDS care. AB - Occupational stress and burnout are potential threats to quality of care for people living with HIV/AIDS. A total of 445 nurses who provide care to people living with HIV/AIDS responded to an anonymous postal survey that consisted of: demographic and work history questions, the Maslach Burnout Inventory, Rotter's Internal-External Locus of Control scale, and internal and external coping styles as measured by the Ways of Coping scale. Path analyses showed that both external (13.6% of the variance) and internal (3.1% of the variance) coping styles significantly predicted levels of burnout among AIDS caregivers, over and above participants' age, perceived workload and locus of control. Mediation analyses further showed that external coping mediates the effect of locus of control on burnout, but internal coping styles do not exhibit mediation. Results therefore replicate and extend previous research demonstrating the important roles of cognitive and behavioural coping styles in understanding burnout among providers of AIDS care. PMID- 10827856 TI - Risk behaviours for HIV and hepatitis infection among anabolic-androgenic steroid users. AB - This study examined HIV and hepatitis risk behaviours among anabolic-androgenic steroid (AAS) users and controls, providing a detailed assessment of both injection and sexual practices. Fifty AAS-using and 40 non-AAS-using weight trainers completed a semi-structured interview followed by a detailed questionnaire. The 'high risk' behaviour of sharing needles and syringes was not current practice among the AAS-using sample, with only one user reporting sharing injecting equipment in the past. However, for some users it was common practice to share multi-dose vials (19%) and to divide drugs using syringes (17%). Both these practices are potential routes for HIV and hepatitis infection. Significantly more AAS users (36%) reported increases in sex drive over the last three months than did the control subjects (5%). More AAS users than controls were engaging in sex with more than one partner while infrequently using condoms. These findings support the maintenance of needle exchange schemes which are essential to continued safe injecting practices among AAS users. The elevated sexual risk behaviour of AAS users could be due to AAS's effect of increasing sex drive or to a 'risk taking' personality trait among AAS users. PMID- 10827857 TI - A scale of optimism-scepticism in the context of HIV treatments. AB - The aim of this research was to develop a brief scale of gay men's optimism scepticism in the context of new HIV treatments. Following comprehensive literature search and communication with other researchers, 34 items about viral load testing, HIV therapies and concern about HIV infection were generated. By way of anonymous, self-complete questionnaires, these items were put to 532 gay men recruited at the Sydney Gay and Lesbian Mardi Gras Fair Day in February 1999. Principal axes factor analysis and item refinement yielded an optimism-scepticism scale based on 12 items (range 12-48, with higher scores indicating greater optimism). The sample mean was 19.8 (SD = 4.7). The scale had high internal consistency reliability (Cronbach's alpha = 0.79). Providing preliminary evidence of validity, men who reported unprotected anal intercourse with a causal partner 'in the past six months' had a significantly higher mean score (21.3) than those who did not report this risk practice (19.5, p < 0.001). Similar validation was provided from separate samples of gay men in Queensland and Sydney. Our scale of optimism-scepticism in the context of new HIV therapies is a brief, reliable instrument which can be incorporated within broader questionnaires and interview schedules. It is a useful tool to monitor attitudes toward HIV therapies and possible associations with risk practices. PMID- 10827858 TI - A review of HIV testing policies and procedures for pregnant women in public maternity units of Porto Alegre, Rio Grande do Sul, Brazil. AB - HIV/AIDS is considered one of the major public health issues in Brazil, with 120,399 notified AIDS cases by 1998 (Aristedes Barbosa et al., 1998). Rio Grande do Sul (RS) reports the highest HIV prevalence in women in the country (Louireiro et al., 1998) and local sero-surveillance studies note an increase in HIV prevalence in pregnancy from 2.6% in 1996 (Buchalter et al., 1996) to 3.3% in 1997 (Ferreira & Valente, 1997). As a result, the Ministry of Health has recommended that all pregnant women are offered an HIV test. This study reviewed HIV testing policies and procedures, reported by obstetric staff in public health care centres in Porto Alegre. The beliefs and practices of obstetric health care professionals towards HIV testing and prevention for pregnant women were also examined, given the crucial role of obstetric staff in the implementation of successful antenatal HIV testing procedures. In total, 106 (69.3%) of eligible obstetric health care professionals responded to a postal questionnaire during April and June 1998. Eighty-six (83.5%) of respondents reported a policy of universal offer of HIV testing. A few reported selective offer (n = 9, 8.7%) and eight (7.8%) reported a policy of testing on request only. When selection was operational, respondents reported aiming at women who disclosed intravenous drug use (88.9%), women whose partner had a history of injecting drugs (77.8%), recipients of blood products prior to 1985 (66.7%), women who reported one or more sexually transmitted diseases (STDs) (88.9%) and women whose current or previous partner ever had sex with a man (77.8%). Respondents reported that consent for an HIV test was acquired 'verbally' by the majority (n = 90, 88.2%), whilst ten (9.8%) acquired consent in writing. Two (2.0%) reported that women's consent for an HIV test was not asked for. Units with universal antenatal HIV testing policy were significantly more likely to have had pregnant women test HIV positive compared with non-universal policy units (U = 13.500, p = 0.050). It appears that whilst the majority of the public health centres were offering an HIV test to all pregnant women in Porto Alegre by 1998, a lack of structured auditing of the actual uptake of HIV testing by pregnant women still remains. Local HIV testing strategy would greatly benefit from such data. PMID- 10827859 TI - The implementation of the DHHS guidelines for the use of antiretroviral agents in HIV-infected adults: a pilot study. AB - The objective of this study was to assess whether United States DHHS guidelines for treatment with antiretroviral therapies are being implemented by health care providers when discussing treatment options with newly diagnosed HIV patients. Health care providers were observed interacting with HIV patients while making decisions about treatment. Observers assessed whether providers and patients discussed the five benefits and six risks of initiating antiretroviral therapy, as recommended in the DHHS Guidelines. Preliminary results indicate that the DHHS Guidelines are not being implemented in the way they were intended. The risks and benefits of antiretroviral therapy were infrequently discussed. No more than four of the 11 risk/benefit items were discussed with any patient, and four of the six risk items were never mentioned to any patient. Potential reasons why the guidelines are not being implemented are discussed. PMID- 10827860 TI - Correlates of high-risk sexual behaviour among Canadian men of South Asian and European origin who have sex with men. AB - A sample of 98 Canadian homosexual and bisexual men, 46 of South Asian and 52 of European origin, who had sex with other than an exclusive primary partner were asked about their high-risk sexual behaviours during the previous six months. They were also queried about internalized homophobia, acculturation to the gay community, and for South Asians acculturation to the majority culture. Participants who reported more internalized homophobia were more likely to engage in both high-risk anal and oral sex. South Asian men exhibited significantly greater levels of homophobia. In addition, South Asian men who were less acculturated to the majority culture were more likely to engage in both types of high-risk sex. These data suggest a need to address internalized homophobia in HIV prevention programmes with homosexual and bisexual men generally, and further suggest the need to target less acculturated South Asian men in particular. PMID- 10827861 TI - Knowledge about HIV/AIDS, the perceived risks of infection and sources of information of Asian-Indian adolescents born in the USA. AB - This study examined the extent and specificity of knowledge about HIV/AIDS, the most used sources of information and the usefulness of these sources among Asian Indian adolescents who were born in the USA and whose parents emigrated from India. Although 86% knew that having unsafe sex with a person infected with HIV could transmit HIV, 47% did not know that sharing a razor with an HIV-positive person could do so, and a significant proportion believed that donating blood (27%) and taking blood tests (14%) could transmit HIV. Television was the most used source of information, but school programmes on HIV/AIDS were considered the most useful source. The results indicated that to be effective, HIV/AIDS prevention programmes must assess the gap in scientific knowledge and beliefs, and clarify misconceptions, reinforce school programmes to present clear messages about the transmission of HIV/AIDS and utilize television to reach adolescents. PMID- 10827862 TI - Modalities of palliative care in hospitalized patients with advanced AIDS. AB - This prospective multidisciplinary survey started in October 1994. The survey assessed the modalities of care of hospitalized patients with advanced AIDS in an Infectious and Tropical Diseases Unit with regards to the practices of palliative care in a Palliative Care Unit. Seventy-eight (78) AIDS patients with CD4 < or = 30/mm3 who had 102 consecutive hospitalizations were recruited. Types (symptomatic or curative) and number of drugs administered to the patients, as well as biological and radiological investigations performed were recorded. Symptoms were concomitantly assessed on a weekly basis by self-evaluation of the patients themselves and by physicians. The results showed that the practices of care were different in the two units according to the specific goals and norms of each unit. A higher density of care was delivered at the Infectious and Tropical Diseases Unit. Symptoms assessed by both patients and physicians were underestimated by physicians in frequency and in intensity. In conclusion, an integrated approach including objective and subjective criteria should enable a better adjustment of the palliative and curative therapeutic strategies in advanced AIDS. These would concomitantly take into account the wishes of the patient and the goals regarding care in the unit where the patient is hospitalized. PMID- 10827863 TI - When always isn't enough: implications of the late application of condoms for the validity and reliability of self-reported condom use. AB - Condoms are most effective when they are applied before intercourse. This study assessed the prevalence of 'condom cheats'--instances of condom use in which the condom is put on after initial penetration. As part of a prospective study of safer sexual behaviour, 103 heterosexual tertiary students completed a condom use diary over a period of up to 6 months. Of the 464 condoms used by study participants, 13% were put on after initial penetration. Thirty-eight percent of condom users reported at least one instance of late application of a condom. Late application of condoms is common and places individuals at risk of infection with HIV and many other STDs. The widespread practice of this behaviour casts doubt on the validity of self-reports of condom use. Assessments of risk based on self reported condom use may lead to underestimates of the risk entailed in heterosexual young adults' condom use behaviour, and may also lead to underestimates of condom efficacy. PMID- 10827865 TI - Seasonality of Plasmodium vivax and P. falciparum in tribal villages in central India (1987-1995). AB - Microscopical examination of blood films produced from samples collected, over a 9-year period (1987-1995), from the inhabitants of four tribal villages of Mandla district, Madhya Pradesh (central India) revealed that malaria was highly endemic and probably transmitted perennially. Both Plasmodium vivax and P. falciparum were prevalent in all age-groups but their prevalence was highly seasonal: longitudinal studies showed an autumn (October-November) peak for P. falciparum and a summer (April-May) peak for P. vivax. However, both the incidence and prevalence of infection with each Plasmodium species showed inter-village variations. Analysis of the malariometric parameters investigated revealed that there had been no improvement in the malaria situation over the study period, and that, since 1992, there had been a shift in the predominant parasite, from P. vivax to P. falciparum, in each village. PMID- 10827864 TI - An evaluation of peer-led STD/HIV prevention work in a public sex environment. AB - This paper describes an independent evaluation of a peer-led STD/HIV prevention intervention conducted by Gay Men Fighting AIDS (GMFA) in a public sex environment (PSE). A variety of quantitative and qualitative research methods were employed to collect data on the intervention process as well as its outcomes. The main aim of the intervention was the distribution of condoms and safer sex literature to PSE users. During a five-month period, over 100,000 condoms and 2,200 safer sex information packs were distributed by GMFA volunteers to the PSE users. Condom provision was identified as the most needed health promotion activity in PSEs in a survey of gay and bisexual men (n = 688) conducted by the evaluators. Data collected showed that condoms provided by GMFA, as well as from other sources, were being used in the PSE. The peer-led focus of the intervention was acceptable to the PSE users. In addition, high levels of commitment and input from the volunteers contributed considerable added value to the intervention. The evaluation found that GMFA was successful in reaching the target population and addressing their needs and demands. PMID- 10827866 TI - Impact of the development of agricultural land on the transmission of sleeping sickness in Daloa, Cote d'Ivoire. AB - Although tools to control sleeping sickness do exist, their use is difficult; areas where intervention is most required often cannot be targeted for lack of appropriate risk indicators. The importance of human behaviour and habits in the manifestation of the disease is clear. In the development of effective new approaches to the control of the disease, information must be gathered about human populations, and their interaction with the environment, in rural as well as in urban and peri-urban areas. The results of a study carried out in Daloa show that use of some methods for the development of agricultural land leads to increased human-vector contact and, as a result, increased risk of sleeping sickness. Such land-management methods may therefore be useful as risk indicators for transmission. Transmission does not occur in the town of Daloa itself but in surrounding areas under cultivation. The use of the epidemiological risk index seems to be inappropriate in urban (and perhaps peri-urban) areas. The results emphasise not only the importance of environmental and demographic data in elucidating the epidemiology of human trypanosomiasis but also the need for further investigations in peri-urban areas. PMID- 10827867 TI - Phylogenetic analysis of the 5' subterminal region of isolates of Leishmania RNA virus-1. AB - Leishmania RNA virus-1 (LRV1) is a double-stranded RNA virus present in some Leishmania species. The virus genome consists of a 450-nucleotide, 5' untranslated region (UTR) followed by the coat gene and the RNA-dependent RNA polymerase (RDRP). It has been shown that the 5' end UTR of the genome promotes internal initiation of translation in an in-vitro assay, indicating the presence of an internal ribosomal entry site (IRES) element upstream of the coat gene. The nucleotide sequences of the 5' subterminal regions of six new isolates of LRV1, of different geographical origins, have now been determined. The RNA folding of the 5' subterminal region of LRV1 has been predicted, using a combination of thermodynamic parameters and folding constraints based on nucleotide substitutions. Furthermore, a putative pyrimidine-rich region (a feature unique to all IRES elements), which is complementary to the Leishmania 18S rRNA, has been identified. The significance and relevance of these findings in the context of the function of the 5' UTR of LRV1 as an IRES element are discussed. PMID- 10827868 TI - Echinococcosis in children and adolescents in Bulgaria: a comparative study. AB - Between 1971 and 1995, 5874 patients underwent surgery in Bulgaria because of cystic echinococcosis caused by Echinococcus granulosus. Of these 5874, 10.6% were children aged < 15 years and 5.25% adolescents aged 15-19 years, giving annual incidences of 1.25 and 2.03/100,000 inhabitants, respectively. Although the annual incidence of surgery for echinococcosis among adults (3.12/100,000) was higher than the combined value for children and adolescents (1.48/100,000), the data indicate that most human infections with E. granulosus occur during childhood and adolescence. In evaluating the epidemiology of echinococcosis or the effectiveness of a control programme, therefore, reductions or increases in the incidence of clinical disease among children and adolescents indicate an improving or worsening situation, respectively. The incidence of surgical treatment for echinococcosis was higher in males than females in all but the youngest subjects (< 5 years) and adults (> 19 years). It was also higher in rural populations than in urban populations, particularly among children and adolescents. Whereas cysts were found more frequently in the lungs of children and adolescents than their livers (51.8% v. 38.3% of the patients), most cysts found in the adults were hepatic (73.5% of patients) and relatively few were in the lungs (14.4% of patients). PMID- 10827869 TI - Epidemiology of HIV and Schistosoma mansoni infections among sugar-estate residents in Ethiopia. AB - Few studies have examined the interaction between schistosomiasis and infection with human immunodeficiency virus (HIV). The overlap between the two infections, and the effect of HIV infection on the egg output and worm load of individuals co infected with Schistosoma mansoni, were therefore investigated in a sugar estate in central Ethiopia. The 1239 subjects were selected by stratified sampling of residents aged 15-54 years. The intensities of infection with S. mansoni were measured as egg output in stools (all subjects) and as the concentration of circulating cathodic antigen (CCA) in urine (a proxy for worm load, measured in 287 subjects). Schistosome infection was detected in 358 subjects [adjusted prevalence (AP) = 31.4%] and HIV infection in 52 (AP = 3.1%). The two infections clustered into different populations of the estate: the schistosome infections were predominantly found in the camps, and primarily affected young people (aged < 20 years) and those working in the field, whereas the HIV epidemic was found in the main village, primarily affecting those aged > 20 years and those who had recently arrived on the estate. Schistosome infection was detected in 348 of the 1187 HIV-negatives (AP = 31.6%) and 10 of the 52 HIV-positives (AP = 25.1%; P > 0.05). Schistosoma mansoni egg output was significantly lower in the HIV positives than in the HIV-negatives (Mann-Whitney test; P = 0.03; ratio of geometric means = 0.74), and remained so after controlling for potential confounders (gender, age, and residence). However, CCA concentrations (i.e. worm loads) were found to be similar for these two groups, after controlling for potential confounders (age, gender, residence, and duration of residence). PMID- 10827870 TI - Observations on the compatibility between Bulinus spp. and Schistosoma haematobium in the Senegal River basin. AB - Snail-infection experiments were carried out with a number of different species and populations of Bulinus and isolates of Schistosoma haematobium. The parasites came from six localities in the Senegal River basin (SRB), in the Lower Valley (Mbodiene), Middle Valley (Podor, Diatar and Nguidjilone), and Upper Valley (Aroundou and Gallade). Isolates of S. haematobium from the Middle and Upper Valleys all showed some compatibility with laboratory-bred B. truncatus from Mali, but none of these isolates was compatible with laboratory-bred B. truncatus originating from Senegal. Schistosoma haematobium from Diatar (Middle Valley) was compatible with B. senegalensis, whereas S. haematobium from Mbodiene (Lower Valley), which is naturally transmitted by B. globosus, was incompatible with B. senegalensis and B. truncatus. These data demonstrate that different isolates of S. haematobium from different regions of the SRB exhibit distinct intermediate host specificities, which in turn will have an effect on the epidemiology of the disease, including the periods of transmission. It is apparent that, in addition to B. senegalensis and B. globosus, B. truncatus, the most widespread bulinid snail in the SRB, may be playing a role in the epidemiology of urinary schistosomiasis. This conclusion has obvious implications for the future spread of urinary schistosomiasis in the SRB. Chemical and physical measurements from assorted habitats along the SRB, including pH, temperature, salinity, conductivity, and resistivity, are also reported. PMID- 10827871 TI - The capacity of the third-stage larvae of Oesophagostomum bifurcum to survive adverse conditions. AB - Human infections with the intestinal nematode Oesophagostomum bifurcum are commonly found in the Sudan savannah of northern Togo and Ghana. Apparently, the long and hot dry season in this region does not prevent transmission, which is believed to take place through ingestion of the infective, third-stage larvae (L3). Oesophagostomum L3 cultured from human stools, unlike the larvae of Necator americanus, were shown to survive desiccation. In addition, 93% of the O. bifurcum L3 frozen for 24 h at -15 degrees C regained motility when brought back into ambient temperatures. The L3 also survived the acidity of an artificial mixture made to resemble the gastric juices of humans. Desiccated larvae could even be rehydrated in this mixture, indicating the possibility of dust-borne infections. The sturdiness of the L3 is likely to contribute to the high transmission intensity in northern Togo and Ghana. PMID- 10827872 TI - Intestinal parasites of the chimpanzee Pan troglodytes in Kibale Forest, Uganda. AB - One-hundred-and-twenty-three stool samples were examined from 45 chimpanzees in a natural population in western Uganda. Comparison with previous studies is complicated by the diversity of techniques used and interpretations. The Ugandan population had relatively many intestinal protozoa, including the probably beneficial entodiniomorph ciliates. Strongyloid nematodes are universal among chimpanzees, but were surprisingly absent from those on Mount Assirik in Senegal. Railletina tapeworms are sporadic in their occurrence. The absence of spiruroid nematodes in the Ugandan population reflects the absence of insectivory in this population. There was little evidence of seasonal difference in prevalences. PMID- 10827873 TI - Leprosy in China: delay in the detection of cases. AB - In a national survey in China, 27,928 cases of leprosy detected by the health authorities between 1984 and 1998 were investigated. The delay between onset of symptoms (estimated from each case's recall) and confirmed diagnosis was < or = 2 years for 55.1% of the new patients but > 10 years for 7.0%, with a median value, overall, of 22.0 months. The median delay was longer: (1) for the multibacillary cases than the paucibacillary; (2) among farmers than among factory workers; (3) among some nationalities than among others (being longest among the Tu and shortest among the Wei); and (4) for some methods of case-detection than for others. Over the study period, the mean delay decreased with time. The delay was greatest in the areas where leprosy was endemic and/or where access to health services was poor. The later the cases were detected the more likely they were to show disability. Leprosy cases are still going undetected in China, although, over the last 14 years, case-finding has significantly improved. Age, occupation, nationality, leprosy type and detection method all appear to affect the delay. PMID- 10827874 TI - An unusual case of multidrug-resistant Plasmodium vivax malaria in Mumbai (Bombay), India. PMID- 10827875 TI - Falciparum malaria: a case report of severe hyponatraemia. PMID- 10827876 TI - Forensic mental health. PMID- 10827877 TI - Developing a clinically useful actuarial tool for assessing violence risk. AB - BACKGROUND: A new actuarial method for violence risk assessment--the Iterative Classification Tree (ICT)--has become available. It has a high degree of accuracy but can be time and resource intensive to administer. AIMS: To increase the clinical utility of the ICT method by restricting the risk factors used to generate the actuarial tool to those commonly available in hospital records or capable of being routinely assessed in clinical practice. METHOD: A total of 939 male and female civil psychiatric patients between 18 and 40 years old were assessed on 106 risk factors in the hospital and monitored for violence to others during the first 20 weeks after discharge. RESULTS: The ICT classified 72.6% of the sample as either low risk (less than half of the sample's base rate of violence) or high risk (more than twice the sample's base rate of violence). CONCLUSIONS: A clinically useful actuarial method exists to assist in violence risk assessment. PMID- 10827878 TI - Evidence-based rehabilitation in forensic psychiatry. AB - BACKGROUND: The extent to which forensic psychiatric rehabilitation alters an individual's level of risk is unclear. AIMS: To highlight some essential features of a forensic psychiatric rehabilitation system, and to discuss risk assessment in this context to create a conceptual framework for risk research and practice. METHOD: The applicability of risk assessment instruments to forensic psychiatric rehabilitation was examined. Core processes and elements considered essential in this type of rehabilitative work were reviewed. RESULTS: Current risk research has limited application to rehabilitation. Future research aimed at analysing forensic psychiatric rehabilitation will be hampered by the complexity of the treatment systems and the number of methodological issues relevant to this type of research. CONCLUSIONS: Novel research approaches are suggested to analyse further the risk factors and processes important in forensic psychiatric rehabilitation. PMID- 10827879 TI - Involuntary out-patient commitment and reduction of violent behaviour in persons with severe mental illness. AB - BACKGROUND: Violent behaviour among persons with severe mental illness (SMI) causes public concern and is associated with illness relapse, hospital recidivism and poor outcomes in community-based treatment. AIMS: To test whether involuntary out-patient commitment (OPC) may help to reduce the incidence of violence among persons with SMI. METHOD: One-year randomised trial of the effectiveness of OPC in 262 subjects with psychotic or major mood disorders and a history of hospital recidivism. Involuntarily hospitalised subjects awaiting OPC were randomly assigned to release or court-ordered treatment after discharge. Those with a recent history of serious assault remained under OPC until expiry of the court order (up to 90 days); then OPC orders were renewed at clinical/court discretion. Control subjects had no OPC. Four-monthly follow-up interviews with subject, case manager and collateral informant took place and service records were collected. RESULTS: A significantly lower incidence of violent behaviour occurred in subjects with > or = 6 months' OPC. Lowest risk of violence was associated with extended OPC combined with regular out-patient services, adherence to prescribed medications and no substance misuse. CONCLUSIONS: OPC may significantly reduce risk of violent behaviour in persons with SMI, in part by improving adherence to medications while diminishing substance misuse. PMID- 10827880 TI - Future directions for treatment in forensic psychiatry. AB - BACKGROUND: As the availability of mental hospital beds has fallen, so the number of people in prison has risen. AIMS: To review current policy trends in British forensic psychiatry and put them in an international context. METHOD: Literature on the prevalence rates of psychiatric disorder in prisons and jails has been examined for the USA, England & Wales and New Zealand. RESULTS: All studies show a high prevalence of mental disorder in prisons and jails. Authors in the USA suggest that prisons are replacing mental hospitals. In England & Wales rates of psychosis are reported as 4-10% for remanded prisoners and 2-7% for sentenced prisoners. Substance misuse among prisoners is a major problem. Prison is the preferred place of disposal for large numbers of mentally disordered people. Does this matter? Why should this be the case? Is this the cheapest option? Politicians are considering new powers to direct more people into institutions (presumably prisons) on the grounds of public protection. CONCLUSIONS: We need more information about attitudes and their formation. We need more interprofessional dialogue about the best arrangements for people with mental disorders, and inter-disciplinary education. PMID- 10827881 TI - Violence in the workplace: new perspectives in forensic mental health services in the USA. AB - BACKGROUND: This paper reviews current research on workplace violence in the USA and offers suggestions concerning the roles that mental health professionals with forensic expertise can play in this expanding field. AIMS: To clarify the role of the mental health professional in evaluating issues related to workplace violence. METHOD: Manual and computer literature searches were performed. RESULTS: The incidence of reported workplace violence is on the rise and can be devastating beyond the immediate injury. Forensically oriented mental health professionals can assist companies by providing pre-employment screenings, fitness-for-duty evaluations and threat assessment by using the results of current research on potentially violent individuals. CONCLUSIONS: With the growing interest in workplace violence come many opportunities for mental health professionals to assist companies in assessment, intervention and prevention. PMID- 10827882 TI - Substance misuse, psychiatric disorder and violent and disturbed behaviour. AB - BACKGROUND: Epidemiological studies suggest schizophrenia and substance misuse to be associated with a higher rate of violence and crime. AIMS: The literature was evaluated to assess whether people with schizophrenia who use substances have an increased risk for violence and disturbed behaviour. METHOD: A detailed Medline analysis was performed and relevant studies were reviewed. RESULTS: A large number of studies have linked substance misuse in schizophrenia with male gender, high incidence of homelessness, more pronounced psychotic symptoms, non-adherence with medication, poor prognosis, violence and aggression. The latter has been proved by clinical, epidemiological and longitudinal prospective studies of unselected birth cohorts. The increased risk for aggression and violent acts cannot be interpreted only as a result of poor social integration. Male gender, more severe psychopathology, a primary antisocial personality, repeated intoxications and non-adherence with treatment are important confounding variables. CONCLUSION: Substance misuse has been shown consistently to be a significant risk factor for violence and disturbed behaviour. Future research should try to evaluate possible pharmacological and psychosocial treatment approaches. PMID- 10827883 TI - Psychiatric morbidity of a rural Indian community. Changes over a 20-year interval. AB - BACKGROUND: Cross-sectional studies give no indication of the changes that may occur in the mental health status of a community in course of times. Studies should be designed to assess these changes. AIMS: To assess the changes, if any, in the prevalence of mental disorders in a rural community after an interval of 20 years in the context of its changing socio-economic conditions. METHOD: A door to-door survey of the prevalence of psychiatric morbidity in two villages was conducted by a team of psychiatrists. The survey was repeated after 20 years by the same team and by the same method. Changes in the mental health status of the community were compared. RESULTS: Total morbidity per 1000 fell from 116.8 to 105.2. Morbidity in men fell from 86.9 to 73.5 per 1000 and in women from 146.8 to 138.3 per 1000. Rates of anxiety, hysteria and phobia had fallen dramatically and those of depression and mania had risen significantly. CONCLUSION: The level of psychiatric morbidity showed no statistically significant change. The morbidity pattern (relative proportion of type of morbidity), however, showed some interesting changes. Similar studies should be done on a larger sample. PMID- 10827884 TI - Catatonia in autistic spectrum disorders. AB - BACKGROUND: The clinical pictures of autistic spectrum disorders include features described in catatonia. AIMS: To examine the severe exacerbation of the catatonic features of autistic disorders in adolescence or early adult life, which occurs in some individuals. METHOD: A semi-structured interview schedule was used to collect information from parents or other care-givers concerning 506 referrals to a specialist clinic for autistic spectrum disorders. Individuals with severe exacerbation of catatonic features were compared with a same-age group of referrals without this type of deterioration in skills and behaviour. RESULTS: Seventeen per cent of referrals aged 15 or over had severe exacerbation of catatonic features. They were significantly more likely than the comparison group to have had, before the onset of the change in behaviour, impaired language and passivity in social interaction. CONCLUSIONS: Catatonia is a later complication of autistic spectrum disorders, which adds considerably to the burden of caring. More research is needed to identify causes, neuropathology, and early signs of vulnerability. PMID- 10827885 TI - Interruption of selective serotonin reuptake inhibitor treatment. Double-blind, placebo-controlled trial. AB - BACKGROUND: Abrupt interruption of therapy with selective serotonin reuptake inhibitors (SSRIs) has been associated with somatic and psychological symptoms. AIMS: Systematically to assess symptoms and effects on daily functioning related to interruption of SSRI therapy. METHOD: Patients treated with fluoxetine, setraline or paroxetine underwent identical five-day periods of treatment interruption and continued active treatment under double-blind, order-randomised conditions, with regular assessment of new symptoms. RESULTS: Placebo substitution for paroxetine was associated with increases in the number and severity of adverse events following the second missed dose, and increases in functional impairment at five days. Placebo substitution for sertraline resulted in less pronounced changes, while interruption of fluoxetine was not associated with any significant increase in symptomatology. CONCLUSIONS: Abrupt interruption of SSRI treatment can result in a syndrome characterised by specific physical and psychological symptoms. Incidence, timing and severity of symptoms vary among SSRIs in a fashion that appears to be related to plasma elimination characteristics. PMID- 10827886 TI - Labour and delivery complications at birth and later mania. An Irish case register study. AB - BACKGROUND: Several reports postulate that manic depression and schizophrenia share environmental risk factors. Although obstetric adversity has been suggested as a risk factor for schizophrenia, few studies have examined its relationship to bipolar affective disorder. AIMS: To assess the rate of obstetric complications incurred by patients with mania compared with controls. METHOD: From the Dublin Psychiatric Case Register we identified individuals with a discharge diagnosis of mania and traced their birth records. Each case was matched with a control of the same gender, born in the same hospital, in the same year, matched for maternal age, parity and social class. Two obstetric complications scales were used to make blind evaluations of labour and delivery data. RESULTS: Patients with mania did not experience a greater frequency or severity of labour and delivery complications than their matched controls. Rates of obstetric adversity were unrelated to the presence or absence of family history of psychiatric disorder. Obstetric adversity was unrelated to the age at first diagnosis. CONCLUSIONS: These findings suggest that obstetric adversity is not a risk factor for later mania. PMID- 10827887 TI - Self-efficacy as a mediator between stressful life events and depressive symptoms. Differences based on history of prior depression. AB - BACKGROUND: Self-efficacy, a characteristic that is protective against depressive symptoms, may be undermined by stressful life events. AIMS: To estimate the effects of stressful life events on self-efficacy, and to examine self-efficacy as a mediator of the effect of stressful life events on symptoms of depression. METHOD: Using a sample of 2858 respondents from the longitudinal Americans' Changing Lives study, path analyses were used to evaluate interrelationships between self-efficacy, life events and symptoms of depression controlling for a variety of potentially confounding variables. Separate models were estimated for those with and without prior depression. RESULTS: For those with prior depression, dependent life events had a significant, negative impact on self efficacy. For those without prior depression, life events had no effect on self efficacy. CONCLUSIONS: For those with prior depression, self-efficacy mediates approximately 40% of the effect of dependent stressful life events on symptoms of depression. PMID- 10827888 TI - Psychometric properties of the Social Phobia Inventory (SPIN). New self-rating scale. AB - BACKGROUND: Of available self-rated social phobia scales, none assesses the spectrum of fear, avoidance, and physiological symptoms, all of which are clinically important. Because of this limitation, we developed the Social Phobia Inventory (SPIN). AIMS: To establish psychometric validation of the SPIN. METHOD: Subjects from three clinical trials and two control groups were given the 17 item, self-rated SPIN. Validity was assessed against several established measures of social anxiety, global assessments of severity and improvement, and scales assessing physical health and disability. RESULTS: Good test-retest reliability, internal consistency, convergent and divergent validity were obtained. A SPIN score of 19 distinguished between social phobia subjects and controls. The SPIN was responsive to change in symptoms over time and reflected different responses to active drugs v. placebo. Factorial analysis identified five factors. CONCLUSIONS: The SPIN demonstrates solid psychometric properties and shows promise as a measurement for the screening of, and treatment response to, social phobia. PMID- 10827889 TI - Subjective imagery in obsessive-compulsive disorder before and after exposure therapy. Pilot randomised controlled trial. AB - BACKGROUND: Distressing mental imagery is hard to study experimentally in obsessive--compulsive disorder (OCD). AIMS: To develop a way to assess mental imagery in OCD during functional magnetic resonance imaging (fMRI). METHOD: A small randomised study, controlled for type and order of mental imagery and for treatment condition (exposure therapy guided by a computer or by a therapist, or relaxation guided by audio-tape). Before and after treatment, during fMRI scanning, patients imagined previously-rehearsed scenarios that evoked an urge to ritualise or non-OCD anxiety or a neutral state, and rated their discomfort during imagery. RESULTS: The method evoked greater discomfort during OCD imagery and anxiety (non-OCD) imagery than during neutral imagery. Discomfort was reduced by cancelling imagery. Discomfort during OCD imagery (but not during anxiety non OCD imagery) fell after exposure therapy but not after relaxation. CONCLUSIONS: Results showed differences between OCD and non-OCD images and their change after successful treatment, and confirmed clinical suggestions that cancelling images reduced OCD discomfort. The method's success paves the way for further studies of mental imagery in OCD: for instance, during fMRI. PMID- 10827890 TI - HoNOS update. PMID- 10827891 TI - Comprehensiveness of systematic review--update. PMID- 10827892 TI - PACT to the future. PMID- 10827893 TI - Drug treatment for resistant depression. PMID- 10827894 TI - Cholesterol, depression and suicide. PMID- 10827895 TI - Cholesterol, depression and suicide. PMID- 10827896 TI - Motor responses to transcranial magnetic stimulation in schizophrenia. PMID- 10827897 TI - Clozapine-induced thrombocytosis. PMID- 10827898 TI - Paternal age and schizophrenia in dizygotic twins. PMID- 10827899 TI - Enhancing the capacity of food consumption surveys of short duration to estimate long-term consumer-only intakes by combination with a qualitative food frequency questionnaire. AB - In principle, a proper risk assessment for a food chemical requires that the time frame for food chemical intake estimates matches the time-frame for the toxicological assessments upon which the safety statements (ADI, PTWI, etc.) are based. For food additives, the toxicological assessments are based on exposure over a lifetime. While food consumption data cannot be collected over the lifetimes of individuals, the information should reflect habitual intakes as closely as possible. This study investigated the possibility of combining a 3-day food diary with a food frequency questionnaire to estimate mean consumer-only food intakes comparable to estimates based on a 14-day diary. The study population consisted of 948 teenagers and analysis was based on 32 clearly defined foods. For 47% of the foods, the difference was < or = 1 g/day. When expressed as portion sizes, 56% of the foods showed differences representing < 5% of an average portion and no food showed a difference > 14% of an average portion. When between-method differences (portions/day) were plotted against the mean of the methods, the mean between-method difference was 0.02(+/- 0.06) portions/day with limits of agreement of -0.10 to 0.14. This preliminary investigation suggests that the combined 3-day diary and FFQ method provides comparable estimates of mean consumer only intakes to a 14-day diary. Therefore, a qualitative FFQ may be a useful adjunct to a food consumption survey of short duration if estimates of longer term food intakes are required. PMID- 10827900 TI - The release of nickel and other trace elements from electric kettles and coffee machines. AB - The release of nickel, chromium and lead from electric kettles to water under conditions simulating regular household use was investigated. Ten out of 26 kettles sold on the Danish market released more than 50 micrograms/l nickel to water, whereas neither lead nor chromium was released in any significant amount. Fifty micrograms/l of nickel in water was chosen as the threshold of action, because concentrations below this value were considered unlikely to provide outbreaks of eczema for those consumers suffering from contact allergy to nickel, who are also sensitive to the content of nickel in the diet. This first part of the study was followed up by a dialogue between the kettle producers and the Danish authorities, leading to a change of construction or design for those kettles that did not comply with the criteria. As a follow-up study another ten kettles were studied to check whether compliance was improved. Two of these ten kettles still released more than 50 micrograms/l nickel to water under the test conditions. These two kettles, however, were subsequently withdrawn from the market. Coffee machines tested similarly did not release aluminium, lead, chromium or nickel in quantities of any significance. PMID- 10827901 TI - Determination of metronidazole and hydroxymetronidazole in trout by a high performance liquid chromatographic method. AB - A high-performance liquid chromatographic (HPLC) method based on solid phase extraction was developed for determination of metronidazole (MNZ) and its metabolite hydroxymetronidazole (MNZ-OH) in muscle and skin tissues of rainbow trout. Tinidazole (TNZ) was used as internal standard. The compounds were extracted with acetonitrile and the extract was evaporated and redissolved in a mixture of ethyl acetate and hexane (1:2). The extract was cleaned up by solid phase extraction on a silica cartridge. The extract was analysed by reverse phase gradient HPLC on a C18 column followed by ultraviolet detection at 325 nm. The limit of detection was 2.8 micrograms/kg for both compounds in muscle. The estimated limits of detection in skin tissue were 3 micrograms/kg for MNZ and 5 micrograms/kg for MNZ-OH. The mean recoveries of MNZ in muscle calculated without use of internal standard were 93% and 81% at levels of 10 micrograms/kg and 25 100 micrograms/kg respectively. The mean recovery of MNZ-OH in muscle was 79% at a level of 10-100 micrograms/kg. The mean relative repeatability standard deviations on spiked muscle tissue were 3.3% for MNZ and 3.2% for MNZ-OH at a level of 10-100 micrograms/kg. The method was applied to trout given feed containing MNZ in an aquaculture pilot plant. Residues of MNZ and MNZ-OH were detected in muscle and skin tissues shortly after the administration period but not 3 weeks later. PMID- 10827902 TI - Estimation of lead intake from crystalware under conditions of consumer use. AB - The purpose of this study was to estimate the lead intake from crystalware resulting from short-term contacts with beverages, under conditions that are likely to occur to a consumer. The extraction ability of different kinds of beverages was estimated by comparison with 4% acetic acid under conditions of continuous contact for 3 h. It was found that lead release increased in the following order: cola drink > HAc > whisky > white wine. Under conditions of repeated use under different scenarios, lead release showed a steep decrease with increasing number of contacts, for both wine and cola drink. The maximum lead intake resulted from the cola drink, corresponding to an ingestion of 14.5 micrograms Pb for consumption of 350 ml beverage. Assuming a fixed contribution from the diet of 71 micrograms/day, in the six scenarios taken into consideration, total daily lead intake levels ranged from a minimum of about 76 micrograms up to a maximum of 86 micrograms lead. As these values, converted on a weekly basis, would correspond to 35% and 40% PTWI respectively, significant health risks resulting from the ingestion of beverages in contact with crystalware can be excluded. Finally it was found that the use of a dishwater did not affect significantly the release of lead into wine, while release into cola drink was slightly but significantly increased after the third cycle. PMID- 10827903 TI - Modification of the ion exchange HPLC procedure for the detection of nitrate and nitrite in dairy products. AB - Existing ion exchange HPLC methodology for nitrate and nitrite analysis in cured meat products suffers from high analyte variability at low concentrations and also chromatographic interference by artifacts in some other foods, such as dairy products. An investigation into the sources of variability has shown that both the cyclohexyl solid phase extraction cartridge and the glass fibre filter used in the original method can introduce artifacts which interfere with the determination of the nitrate in foodstuffs. We have also found that the use of a graphitized solid phase extraction cartridge used in tandem with the cyclohexyl solid phase extraction cartridge removed the artifacts from the chromatograph of dairy products that co-eluted with nitrite and nitrate. Values for the nitrite and nitrate content of dairy products were obtained by the HPLC procedure using these two solid phase extraction cartridges and the values obtained were in close agreement with those obtained by cadmium column reduction and colorimetry. PMID- 10827904 TI - Clinical incidents and risk management--a public health issue. PMID- 10827905 TI - Declining male births with increasing geographical latitude in Europe. AB - OBJECTIVE: Demographic studies in various industrialised countries have shown a decline in male births in the latter half of the 20th century from the expected ratio of 0.515 (males/total). This study analyses trends in this ratio over the period 1890 to 1995 in Malta, and also analyses this ratio for Western European countries for the period 1990-1995. DESIGN: Births subdivided by sex were obtained from official Maltese publications. European countries were grouped according to geographical latitude by banding countries into three groups: Northern Mediterranean, Central European and Scandinavian. Births by sex for these countries were also analysed for the period 1990-1995. RESULTS: No decline in the ratio of male births to total births was noted in Malta over the period 1916-1995. However, the ratio was higher than expected (n = 151,766, ratio = 0.517 (95% confidence intervals (95% CI): 0.514, 0.519). Moreover, during the period 1890-1899 (n = 66,874), the ratio was 0.523 (95% CI: 0.519, 0.527), even higher than observed during the 20th century (chi 2 = 8.3, p = 0.004). Analysis of European births showed a much higher ratio of male births in the south of Europe than in the north (chi 2 = 87.2, p < 0.0001). CONCLUSIONS: The findings were unable to explain the higher incidence of male births in the south of Europe, but it is speculated that ambient temperatures may not only affect fertility, but also influence sex ratios at birth. PMID- 10827906 TI - "Development" is not essential to reduce infant mortality rate in India: experience from the Ballabgarh project. AB - BACKGROUND: India aims to reduce the infant mortality rate (IMR) to below 60 per 1000 live births by 2000. IMR is higher in northern India as compared with south Indian states like Kerala. Any further reduction in IMR needs identification of new strategies. The Ballabgarh project with an IMR of 36 in 1997 can help identify such strategies. OBJECTIVE: To see the trend in reduction of neonatal mortality rate (NNMR) and IMR at the Ballabgarh project, compare it with Kerala and rural India's trend and look at the causes of neonatal and infant mortality. DESIGN: The Comprehensive Rural Health Services Project, Ballabgarh, run by the All India Institute of Medical Sciences, covered an estimated population of 70,079 in 1997. The health care delivery system is on the national pattern. All the deaths are identified during the house visits by the male workers. The cause of death is ascertained by the health assistant based on the symptomatology at the time of death. RESULTS: The trends in reduction of IMR for Ballabgarh, Kerala and rural India are roughly parallel with the IMR of Ballabgarh lying somewhere in between the two. However, the NNMR of Ballabgarh (10.6 in 1996) was comparable to Kerala's NNMR (10.9 in 1992). The proportion of infant deaths occurring during the neonatal period had fallen from 50% in the early seventies to 30% during 1996 97. In 1992-1994, 33.8% of all neonatal deaths were attributable to low birth weight and 37.3% to infective causes. Acute respiratory infection and diarrhoea continue to be the chief cause of postneonatal mortality. CONCLUSION: It is possible to bring down neonatal mortality before postneonatal mortality. The Kerala model, which focuses on social development, may not apply to northern India for sociocultural reasons. PMID- 10827907 TI - Marital status and suicide in the National Longitudinal Mortality Study. AB - OBJECTIVES: The purpose of the study was to examine the effect of marital status on the risk of suicide, using a large nationally representative sample. A related objective was to investigate the association between marital status and suicide by sex. METHODS: Cox proportional hazards regression models were applied to data from the National Longitudinal Mortality Study, based on the 1979-1989 follow up. In estimating the effect of marital status, adjustments were made for age, sex, race, education, family income, and region of residence. RESULTS: For the entire sample, higher risks of suicide were found in divorced than in married persons. Divorced and separated persons were over twice as likely to commit suicide as married persons (RR = 2.08, 95% confidence intervals (95% CI) 1.58, 2.72). Being single or widowed had no significant effect on suicide risk. When data were stratified by sex, it was observed that the risk of suicide among divorced men was over twice that of married men (RR = 2.38, CI 1.77, 3.20). Among women, however, there were no statistically significant differentials in the risk of suicide by marital status categories. CONCLUSIONS: Marital status, especially divorce, has strong net effect on mortality from suicide, but only among men. The study showed that in epidemiological research on suicide, more accurate results would be obtained if samples are stratified on the basis of key demographic or social characteristics. The study further observed that failure to control for relevant socioeconomic variables or combining men and women in the same models could produce misleading results. PMID- 10827908 TI - The cultural heritage shapes the pattern of tumour profiles in Europe: a correlation study. AB - STUDY OBJECTIVE: This study investigates the spatial pattern of tumours in Europe to check the feasibility of a large scale ecological epidemiology approach to cancer in Europe. SETTING: The tumour types relative frequencies and cancer incidence (for men and women) reported in the European cancer registries were investigated by exploratory data analysis techniques. Socioeconomical descriptors of the female condition were considered as well. MAIN RESULTS: The classification of the European regional areas covered by the cancer registries followed almost exactly the boundaries set by the long and intermingled European history in terms of life styles and cultural heritage. This result supports the notion of a predominant role of environmental factors in cancer induction. Further support to the above result was given by the finding of a correlation between differential male-female cancer incidence, and socioeconomic descriptors of the female condition. CONCLUSIONS: From a methodological point of view, the consistency of these results pointed to the feasibility of an ecological approach to tumour epidemiology. PMID- 10827909 TI - Sociodemographic characteristics influencing birth outcome in Sweden, 1908-1930. Birth variables in the Population Study of Women in Gothenburg. AB - STUDY OBJECTIVE: To identify variables available in early Swedish delivery records and their relation to birth outcomes for home and hospital deliveries in Gothenburg at the early part of this century. DESIGN: A retrospective recovery of original delivery records and social variables in a cross sectional population. SETTING: Gothenburg, Sweden. PARTICIPANTS: 851 fullterm singleton female births with known gestational age born into five birth cohorts on selected dates (1908, 1914, 1918, 1922 and 1930). MAIN RESULTS: Delivery site, maternal parity, gestational age, and social group were significant factors influencing birth outcome as birth weight and length. The mean birth weight and length of hospital born infants was consistently lower than for home deliveries across all cohorts. Site of delivery changed significantly during the period of births under study, 1908-1930. CONCLUSIONS: In this study, which was based on original delivery records from the early part of this century, it was found that delivery site was an important factor influencing birth outcome across five birth cohorts. Utilisation of delivery services changed during the period of study. Thus, to avoid selection bias, the application of delivery records should reflect the birthing practice of the time period in question. PMID- 10827910 TI - Intra-urban differences in breast cancer mortality: a study from the city of Malmo in Sweden. AB - STUDY OBJECTIVE: To assess whether in an urban population stage at breast cancer diagnosis is related to area of living and to what extent intra-urban differences in breast cancer mortality are related to incidence respectively stage at diagnosis. DESIGN: National registries were used to identify cases. Mortality in 17 residential areas was studied in relation to incidence and stage distribution using linear regression analysis. Areas with high and low breast cancer mortality, incidence and proportion of stage II+ tumours at diagnosis were also compared in terms of their sociodemographic profile. SETTING: City of Malmo in southern Sweden. PATIENTS: The 1675 incident breast cancer cases and 448 deaths that occurred in women above 45 years of age in Malmo 1986-96. MAIN RESULTS: Average annual age standardised breast cancer mortality ranged between residential areas, from 35/10(5) to 107/10(5), p = 0.04. Mortality of breast cancer was not correlated to incidence, r = 0.22, p = 0.39. The ratio of stage II+/0-I cancer incidence varied between areas from 0.45 to 1.99 and was significantly correlated to breast cancer mortality, r = 0.53, p = 0.03. Areas with high proportion of stage II+ cancers and high mortality/incidence ratio were characterised by a high proportion of residentials receiving income support, being foreigners and current smokers. CONCLUSIONS: Within this urban population there were marked differences in breast cancer mortality between residential areas. Stage at diagnosis, but not incidence, contributed to the pattern of mortality. Areas with high proportion of stage II+ tumours differed unfavourably in several sociodemographic aspects from the city average. PMID- 10827911 TI - Socioeconomic status and the expectation of disability in old age: estimates for England. AB - OBJECTIVES: The longer life expectancy in old age of more privileged socioeconomic groups is well established, but less clear is whether the net effect of additional years of life is a lengthened, stable or reduced duration of disability. Estimates of healthy and disabled life expectancy (using definitions including dependency in activities of daily living and cognitive impairment) were made, contrasting occupational classes I and II (professional and managerial) with the rest. DESIGN: Disability prevalence was estimated from the Medical Research Council Cognitive Function and Ageing study. Sullivan's method was used to calculate health expectancy. SUBJECTS: 10,377 people aged 65 years or over in Cambridgeshire, Newcastle, Nottingham and Oxford. Subjects were classified as disabled if they had evidence of dementia (using the Automated Geriatric Examination Computer Assisted Taxonomy) or scored 11 or more on the modified Townsend Disability scale, at baseline screen. RESULTS: The prevalence of disability overall and need for "constant care" was lower in both men and women in social classes I and II compared with the rest. Men aged 65 to 69 in classes I and II can expect nearly 14 years of life free of disability compared with 11.5 years for those in classes III to V: for women the equivalent expectations are 15.5 and 13.8 years. Men aged 65 to 69 in classes I and II can also expect a shorter duration of disability: 1 year compared with 1.6 years for classes III to V. In women expectation of disability is higher overall, but shows little difference by occupational class. CONCLUSIONS: Relatively privileged socioeconomic groups in England, especially men, can expect fewer years of disability despite longer overall life expectancy. These findings lend weight to optimistic scenarios for the future numbers of older people with disability. PMID- 10827912 TI - Regional variation in incidence and case fatality of myocardial infarction among young women in England, Scotland and Wales. AB - OBJECTIVES: To examine the regional variation in incidence and case fatality of myocardial infarction among young women. DESIGN: Cross sectional survey, using population based incidence data. SETTING: England, Scotland and Wales. SUBJECTS: Subjects were women aged 16-44 with a diagnosis of myocardial infarction between 1 October 1993 and 15 October 1995. OUTCOME MEASURES: Incidence of myocardial infarction per 100,000 women years, with case fatality as a percentage of total cases. RESULTS: Incidence of myocardial infarction rose steeply from age 33 upwards, (maximum = 20.2 cases per 100,000 women years at age 44). The adjusted incidence rate for myocardial infarction was 3.7 (95% CI 3.2, 4.2) times greater in Scotland than in southern England. In contrast, case fatality was significantly lower in Scotland: 18.5% (95% CI 13.1%, 25.0%), compared with 31.0% (95% CI 25.9%, 36.0%) in southern England. CONCLUSIONS: The incidence of myocardial infarction varied widely within the United Kingdom. Case fatality variation may reflect differences in ambulance response, or in diagnostic acumen, within the regions. PMID- 10827913 TI - Income, occupational position, qualification and health inequalities--competing risks? (comparing indicators of social status). AB - STUDY OBJECTIVE: The debate on health inequalities has shifted from the consequences of occupational position, as expressed in the Registrar General's classification, to consequences of material living conditions. This change in interest occurred without comparative analyses of different sources of health inequalities. Thus this study investigated the relative contribution of "material resources" (income), "qualification" and "occupational position" for explaining social differentials in mortality. DESIGN AND SETTING: Analyses were performed with records from a statutory health insurance in West Germany. The analyses were performed with data of 84,814 employed men and women between 25 and 65 years of age who were insured between 1987 and 1995 for at least 150 days. RESULTS: The three indicators were statistically associated, but not strong enough to warrant the conclusion that they share the same empirical content. The relative risk (hazard rate) for income by controlling for occupational position and gender for the highest as compared with the lowest category was 1.99 (95% CI 1.66, 2.39). The corresponding relative risk for income by controlling for qualification and gender was 2.03 (95% CI 1.68, 2.46). In both multivariate analyses, the effects of occupational position and qualification were no longer interpretable because of large confidence intervals. In sum, income related relative mortality risks were the comparably highest, while qualification and occupational position were no longer substantial. CONCLUSIONS: The results emphasise the present discussion on the consequences of material living conditions. Income on the one hand and qualification and occupational position on the other are largely independent. Mortality related effects of income override those of the other socioeconomic status indicators. However, seen in a time perspective, qualification may still have a placement function at least for the first occupational position. PMID- 10827914 TI - The impact of area deprivation on differences in health: does the choice of the geographical classification matter? AB - OBJECTIVE: Many studies show the average health status in deprived areas to be poorer and the use of health care to be higher, but there is hardly any information on the impact of the geographical classification on the size of these differences. This study examines the impact of the geographical classification on the clustering of poor health per area and on the size of the differences in health by area deprivation. DESIGN: Data on self reported health regarding 5121 people were analysed using three classifications: neighbourhoods, postcode sectors and boroughs. Multilevel logistic models were used to determine the clustering of poor health per area and the size of the differences in health by area deprivation, without and subsequently with adjustment for individual socioeconomic status. SETTING: General population aged 16 years and over of Amsterdam, The Netherlands. MAIN OUTCOME MEASURES: Self rated health, mental symptoms (General Health Questionnaire, 12-item version), physical symptoms and long term functional limitations. MAIN RESULTS: The clustering of poor health is largest in neighbourhoods and smallest in postcode sectors. Health differences by area deprivation differ only slightly for the three geographical classifications, both with and without adjustment for individual socioeconomic status. CONCLUSIONS: In this study, the choice of the geographical classification affects the degree of clustering of poor health by area but it has hardly any impact on the size of health differences by area deprivation. PMID- 10827915 TI - Balkan briefing (Part 4). Kosovo refugees in the North West region of the United Kingdom. PMID- 10827916 TI - What is the lag time between income inequality and health status? PMID- 10827917 TI - Aetiological heterogeneity of Alzheimer's disease. PMID- 10827918 TI - A survey of smoking among Italian doctors. PMID- 10827923 TI - Recent advances in the preparation of heterocycles on solid support: a review of the literature. PMID- 10827924 TI - Combinatorial de novo synthesis of catalysts: how much of a hit-structure is needed for activity? PMID- 10827925 TI - String synthesis. A spatially addressable split procedure PMID- 10827926 TI - Kinetic study of organic reactions on polystyrene grafted MicroTubes PMID- 10827927 TI - Resin-to-resin suzuki coupling of solid supported arylboronic acids PMID- 10827928 TI - Photoinduced cyclorelease for condensed heteroaromatic synthesis. PMID- 10827929 TI - Synthesis of non-peptidyl alpha,alpha-difluoromethylenephosphonic acids on a soluble polymer support AB - The polymer-supported syntheses of a series of biaryl derivatives bearing the alpha,alpha-difluoromethylenephosphonic acid group is reported. Non-cross-linked polystyrene (NCPS) was used as the support which enabled the reactions to be carried out under homogeneous conditions and reactions to be followed using conventional (19)F NMR. Synthesis of the biaryl phosphonic acids was initiated by attaching mono-ethyl esters of alpha, alpha-difluorophosphonic acids 11 and 12 to 3% alkylhydroxy-modified NCPS via a phosphate ester linkage. Suzuki reaction conditions were developed which allowed for the formation of a series of polymer bound biaryl phosphonates at ambient temperature. Removal of phosphonic acids from the support and cleavage of the ethyl protecting group was achieved in a single step using TMSI or TMSBr. Yields of the phosphonic acids ranged from 43 to 89% and, in most cases, were obtained in a purity (96-99%), after cleavage from the support, that was sufficient for biological screening. PMID- 10827930 TI - Chemically sensitive high throughput parallel analysis of solid phase supported library members AB - This paper introduces Fourier transform infrared imaging as a powerful spectroscopic tool for the parallel identification of members of resin-supported combinatorial libraries. This technique combines the chemical specificity and high sensitivity of FTIR with the ability to rapidly analyze multiple supported resin beads simultaneously. It is shown here that the chemical identity of ligands on a variety of supported resin beads can be identified in a single experiment without destroying or otherwise perturbing the system. PMID- 10827931 TI - A new cleavage strategy for ester linked polymer supports: generation of a tertiary alcohol library AB - The addition of different Grignard reagents to polymer bound esters resulting in the generation of a tertiary alcohol library with concomitant cleavage is described. PMID- 10827932 TI - Solid phase synthesis of purines from pyrimidines. AB - In this paper the solid phase synthesis of various substituted purines is described starting from 4,6-dichloro-5-nitropyrimidine. The 4,6-dichloro-5 nitropyrimidine was coupled to Rink amide resin followed by displacement of the second chloride by an amino compound. Reduction of the nitro compound proved to be problematic but was achieved using lithium aluminum hydride/aluminum trichloride. The diamines (13) were then elaborated to purines by three different routes. PMID- 10827933 TI - Evaluation of evaporative light-scattering detector for combinatorial library quantitation by reversed phase HPLC. AB - A quantitation study using reversed phase HPLC with UV and evaporative light scattering detector (ELSD) was conducted on 90 library standards selected from 15 small molecule combinatorial libraries (six standards from each library). This study assessed the quantitation errors using a single calibration curve for rapid purity analysis of combinatorial libraries. The average quantitation error of six standards from one library at 200 microM by UV was 13. 4%, 20.6%, and 60.3%, at 214, 220, and 254 nm, respectively. By ELSD, the average quantitation error of these six standards at 200 micro was only 7.7%. Applying this ELSD calibration curve to 84 standards from 14 structurally diverse libraries, an average quantitation error of 16.4% was obtained. The average quantitation error of all 90 standards from 15 libraries using 15 calibration curves was 18.5%. PMID- 10827934 TI - Confirmation of primary active substances from high throughput screening of chemical and biological populations: a statistical approach and practical considerations. AB - Many biologically important substances are discovered through screening of relevant chemical or biological libraries. The ability to find the active substances ("hits") from any random collection is largely determined by the quality of the assay and screening conditions. When a large population is screened for a specific characteristic, each member of that population is usually tested only once. Errors in the measurements require additional follow-up tests to confirm that each hit from the primary screen is truly active. In this report, we present a statistical model system that predicts the reliability of hits from a primary test as affected by the error in the assay and the choice of the hit threshold (hit limit). The hit confirmation rate, as well as false positive (representing substances that initially fall above the hit limit but whose true activity are below the hit limit) and false negative (representing substances that initially fall below the hit limit but whose true activity are in fact greater than the hit limit) rates have been analyzed with this model by computational simulation. This model can also be used in screen validation and post-screening data analysis. The statistical analysis presented here has broad implications and is applicable to screening of any large population for any specific characteristic. Obvious applications include drug discovery, gene chip analysis, population biology, directed molecular evolution, biological panning, and combinatorial material sciences. PMID- 10827935 TI - Application of base cleavable safety catch linkers to solid phase library production. AB - We have used sulfide "Safety Catch" linkers to anchor typical medicinal chemistry functional groups to amine resins. Compounds are loaded as the ester, carbamate, or amine. At the end of the synthesis, the linker is activated by peracid. The sulfone resins are then cleaved by beta-elimination in the gas phase or in solution by secondary amines to produce acids and primary, secondary, or tertiary amines. Comparison of cleavage rates to other sulfone resins including SEM showed significantly faster cleavage for this system with conditions similar to Fmoc deprotection. Application of this strategy to a medicinal chemistry library gives good yields and purities of the resulting compounds. PMID- 10827936 TI - Solution- and solid-phase syntheses of substituted guanidinocarboxylic acids. AB - A library of guanidine-based compounds was produced to mimic the lead compound 1, which is a substance known to have intensely sweet-taste characteristics. Libraries of guanidinocarboxylic acids were therefore prepared via two synthetic methods. The solid-phase method involving trapping of solution-phase carbodiimides by supported amines was used to produce N,N'-dialkyl derivatives (Scheme 1). The second solid-phase method, featuring supported carbodiimides and solution-phase amines (Scheme 2), was devised to prepare N,N'-disubstituted and N,N',N'-trisubstituted guanidinocarboxylic acids. A small collection of guanadinoacetic acid dimers and trimers was also prepared, but this time via a solution-phase coupling of carbodiimides to a polyamine linker. PMID- 10827937 TI - Solid-phase syntheses of heterocycles containing the 2-aminothiophenol moiety. AB - Efficient and general procedures have been developed for the solid-phase preparation of substituted benzothiazoles (1), 3, 4-dihydro-1,4-benzothiazines (2), 3,4-dihydro-1,4-benzothiazine-1, 1-dioxides (3), 3,4-dihydro-3-oxo-1,4 benzothiazines (4), and 3, 4-dihydro-3-oxo-1,4-benzothiazine-1,1-dioxides (5). All five classes of compounds were prepared from a common intermediate, resin bound 2-amino-4-carboxythiophenol, in a minimal number of steps. This intermediate was generated by (i) coupling 4-fluoro-3-nitrobenzoic acid onto Wang resin, or onto an amino acid bound to the resin, (ii) substitution of the aryl fluoride with a protected thiol, (iii) reduction of the nitro group, and (iv) removal of sulfur protection. Reaction with the appropriate substrates and reagents to effect cyclization gave the substituted core structures, which were modified further to introduce additional point(s) of diversity. Following cleavages from the solid support, the compounds were obtained in high initial purities and good isolated yields after purification. PMID- 10827938 TI - SPEEDY: spin-echo enhanced diffusion filtered spectroscopy. A new tool for high resolution MAS NMR AB - The pulsed field gradient diffusion edited experiment, bipolar LED, has been combined with the Carr-Purcell-Meiboom-Gill (CPMG) spin-echo sequence for the analysis of solid phase resin samples. Spin-echo enhanced diffusion filtered spectroscopy (SPEEDY), when optimized, filters both the compounds that demonstrate fast diffusion rates as well as the compounds that demonstrate fast T(2) relaxation rates. Using this technique, compounds that are not covalently attached to the resin are not observed and contributions from the resin matrix are greatly attenuated. The interpretation of the resulting spectrum is more readily accessible. This technique lessens the importance of completely removing reaction residues or the wash solvents simply for analytical evaluation. The utility of the combined filtering scheme was demonstrated by the implementation into a NOESY sequence. PMID- 10827939 TI - Gatekeepers of the nucleus. AB - Nuclear pore complexes (NPCs) form the site for entry and exit from the nucleus. A convergence of studies have defined the physical framework for the nuclear transport mechanism. This includes definition of the soluble transport machinery required for protein and RNA movement, x-ray structure analysis of transport factors, definitive compositional analysis of yeast NPCs, and documentation of the relative steady state arrangement of NPC components within the portal. With this information, researchers are now in the exciting position to examine the dynamic interplay between shuttling transport factors and the static pore complex. PMID- 10827940 TI - A sense of the end. AB - How a cell distinguishes a double-strand break from the end of a chromosome has long fascinated cell biologists. It was thought that the protection of chromosomal ends required either a telomere-specific complex or the looping back of the 3' TG-rich overhang to anneal with a homologous double-stranded repeat. These models must now accommodate the findings that complexes involved in nonhomologous end joining play important roles in normal telomere length maintenance, and that subtelomeric chromatin changes in response to the DNA damage checkpoint. A hypothetical chromatin assembly checkpoint may help to explain why telomeres and the double-strand break repair machinery share essential components. PMID- 10827941 TI - Splitting the chromosome: cutting the ties that bind sister chromatids. AB - In eukaryotic cells, sister DNA molecules remain physically connected from their production at S phase until their separation during anaphase. This cohesion is essential for the separation of sister chromatids to opposite poles of the cell at mitosis. It also permits chromosome segregation to take place long after duplication has been completed. Recent work has identified a multisubunit complex called cohesin that is essential for connecting sisters. Proteolytic cleavage of one of cohesin's subunits may trigger sister separation at the onset of anaphase. PMID- 10827942 TI - Like attracts like: getting RNA processing together in the nucleus. AB - Structures visible within the eukaryotic nucleus have fascinated generations of biologists. Recent data show that these structures form in response to gene expression and are highly dynamic in living cells. RNA processing and assembly require many factors but the nucleus apparently lacks any active transport system to deliver these to the RNAs. Instead, processing factors move by diffusion but are concentrated by transient association with functionally related components. At sites of high activity this gives rise to visible structures, with components in dynamic equilibrium with the surrounding nucleoplasm. Processing factors are recruited from this pool by cooperative binding to RNA substrates. PMID- 10827943 TI - Structure of the light-driven chloride pump halorhodopsin at 1.8 A resolution. AB - Halorhodopsin, an archaeal rhodopsin ubiquitous in Haloarchaea, uses light energy to pump chloride through biological membranes. Halorhodopsin crystals were grown in a cubic lipidic phase, which allowed the x-ray structure determination of this anion pump at 1.8 angstrom resolution. Halorhodopsin assembles to trimers around a central patch consisting of palmitic acid. Next to the protonated Schiff base between Lys(242) and the isomerizable retinal chromophore, a single chloride ion occupies the transport site. Energetic calculations on chloride binding reveal a combination of ion-ion and ion-dipole interactions for stabilizing the anion 18 angstroms below the membrane surface. Ion dragging across the protonated Schiff base explains why chloride and proton translocation modes are mechanistically equivalent in archaeal rhodopsins. PMID- 10827944 TI - Anomalous polarization profiles in sunspots: possible origin of umbral flashes AB - We present time-series spectropolarimetric observations of sunspots in the Ca II infrared triplet lines, which show a periodic occurrence of anomalous, asymmetric, circular polarization profiles in the umbral chromosphere. The profiles may be caused by the periodic development of an unresolved atmospheric component in a downward flowing magnetized environment. This active component with upward directed velocities as high as 10 kilometers per second is connected to the umbral flash (UF) phenomenon. We can explain the observations with a semiempirical model of the chromospheric oscillation and of the sunspot magnetized atmospheric plasma during a UF event. PMID- 10827945 TI - Dimensionality effects in the lifetime of surface states AB - A long-standing discrepancy between experimental and theoretical values for the lifetimes of holes in the surface-state electron bands on noble metal surfaces is resolved; previous determinations of both are found to have been in error. The ability of the scanning tunneling microscope to verify surface quality before taking spectroscopic measurements is used to remove the effects of defect scattering on experimental lifetimes, found to have been a significant contribution to prior determinations. A theoretical treatment of inelastic electron-electron scattering is developed that explicitly includes intraband transitions within the surface state band. In our model, two-dimensional decay channels dominate the electron-electron interactions that contribute to the hole decay and are screened by the electron states of the underlying three-dimensional electron system. PMID- 10827946 TI - Real-time observation of adsorbate atom motion above a metal surface AB - The dynamics of cesium atom motion above the copper(111) surface following electronic excitation with light was studied with femtosecond (10(-15) seconds) time resolution. Unusual changes in the surface electronic structure within 160 femtoseconds after excitation, observed by time-resolved two-photon photoemission spectroscopy, are attributed to atomic motion in a copper-cesium bond-breaking process. Describing the change in energy of the cesium antibonding state with a simple classical model provides information on the mechanical forces acting on cesium atoms that are "turned on" by photoexcitation. Within 160 femtoseconds, the copper-cesium bond extends by 0.35 angstrom from its equilibrium value. PMID- 10827947 TI - Electron-induced inversion of helical chirality in copper complexes of N,N dialkylmethionines. AB - Stereodynamic complexes of copper were found to undergo inversion of a helical chiral element upon oxidation or reduction. The amino acid methionine was derivatized by the attachment of two chromophores to the nitrogen atom. The resultant ligands formed stable complexes with Cu(I) and Cu(II) salts. For a derivative of a given absolute chirality, the complexes afford nearly mirror image circular dichroism spectra. The spectral changes originate from reorientation of the nitrogen-attached chromophores due to a conformation interconversion driven by the exchange of a carboxylate for a sulfide ligand. The electrically induced chirality inversion coupled with strong interactions with polarized light is unique and may lead to novel chiral molecular devices. PMID- 10827948 TI - Quantifying denitrification and its effect on ozone recovery AB - Upper Atmosphere Research Satellite observations indicate that extensive denitrification without significant dehydration currently occurs only in the Antarctic during mid to late June. The fact that denitrification occurs in a relatively warm month in the Antarctic raises concern about the likelihood of its occurrence and associated effects on ozone recovery in a colder and possibly more humid future Arctic lower stratosphere. Polar stratospheric cloud lifetimes required for Arctic denitrification to occur in the future are presented and contrasted against the current Antarctic cloud lifetimes. Model calculations show that widespread severe denitrification could enhance future Arctic ozone loss by up to 30%. PMID- 10827949 TI - Field measurement of slow metamorphic reaction rates at temperatures of 500 degrees to 600 degrees C AB - High-temperature metamorphic reaction rates were measured using strontium isotopic ratios of garnet and whole rock from a field site near Simplon Pass, Switzerland. For metamorphic conditions of cooling from 612 degrees +/- 17 degrees C to 505 degrees +/- 15 degrees C at pressures up to 9.1 kilobars, the inferred bulk fluid-rock exchange rate is 1.3(-0.4)(+1.1) x 10(-7) grams of solid reacted per gram of solid per year, several orders of magnitude lower than laboratory-based estimates. The inferred reaction rate suggests that mineral chemistry may lag the evolving conditions in Earth's crust during mountain building. PMID- 10827950 TI - Spatial patterns in the distribution of tropical tree species. AB - Fully mapped tree census plots of large area, 25 to 52 hectares, have now been completed at six different sites in tropical forests, including dry deciduous to wet evergreen forest on two continents. One of the main goals of these plots has been to evaluate spatial patterns in tropical tree populations. Here the degree of aggregation in the distribution of 1768 tree species is examined based on the average density of conspecific trees in circular neighborhoods around each tree. When all individuals larger than 1 centimeter in stem diameter were included, nearly every species was more aggregated than a random distribution. Considering only larger trees (>/= 10 centimeters in diameter), the pattern persisted, with most species being more aggregated than random. Rare species were more aggregated than common species. All six forests were very similar in all the particulars of these results. PMID- 10827951 TI - Mechanism of ATP-dependent promoter melting by transcription factor IIH. AB - We show that transcription factor IIH ERCC3 subunit, the DNA helicase responsible for adenosine triphosphate (ATP)-dependent promoter melting during transcription initiation, does not interact with the promoter region that undergoes melting but instead interacts with DNA downstream of this region. We show further that promoter melting does not change protein-DNA interactions upstream of the region that undergoes melting but does change interactions within and downstream of this region. Our results rule out the proposal that IIH functions in promoter melting through a conventional DNA-helicase mechanism. We propose that IIH functions as a molecular wrench: rotating downstream DNA relative to fixed upstream protein-DNA interactions, thereby generating torque on, and melting, the intervening DNA. PMID- 10827952 TI - Structure and function of a human TAFII250 double bromodomain module. AB - TFIID is a large multiprotein complex that initiates assembly of the transcription machinery. It is unclear how TFIID recognizes promoters in vivo when templates are nucleosome-bound. Here, it is shown that TAFII250, the largest subunit of TFIID, contains two tandem bromodomain modules that bind selectively to multiply acetylated histone H4 peptides. The 2.1 angstrom crystal structure of the double bromodomain reveals two side-by-side, four-helix bundles with a highly polarized surface charge distribution. Each bundle contains an Nepsilon acetyllysine binding pocket at its center, which results in a structure ideally suited for recognition of diacetylated histone H4 tails. Thus, TFIID may be targeted to specific chromatin-bound promoters and may play a role in chromatin recognition. PMID- 10827953 TI - Rapid destruction of human Cdc25A in response to DNA damage. AB - To protect genome integrity and ensure survival, eukaryotic cells exposed to genotoxic stress cease proliferating to provide time for DNA repair. Human cells responded to ultraviolet light or ionizing radiation by rapid, ubiquitin- and proteasome-dependent protein degradation of Cdc25A, a phosphatase that is required for progression from G1 to S phase of the cell cycle. This response involved activated Chk1 protein kinase but not the p53 pathway, and the persisting inhibitory tyrosine phosphorylation of Cdk2 blocked entry into S phase and DNA replication. Overexpression of Cdc25A bypassed this mechanism, leading to enhanced DNA damage and decreased cell survival. These results identify specific degradation of Cdc25A as part of the DNA damage checkpoint mechanism and suggest how Cdc25A overexpression in human cancers might contribute to tumorigenesis. PMID- 10827954 TI - Chromatin-independent nuclear envelope assembly induced by Ran GTPase in Xenopus egg extracts. AB - The nuclear envelope (NE) forms a controlled boundary between the cytoplasm and the nucleus of eukaryotic cells. To facilitate investigation of mechanisms controlling NE assembly, we developed a cell-free system made from Xenopus laevis eggs to study the process in the absence of chromatin. NEs incorporating nuclear pores were assembled around beads coated with the guanosine triphosphatase Ran, forming pseudo-nuclei that actively imported nuclear proteins. NE assembly required the cycling of guanine nucleotides on Ran and was promoted by RCC1, a nucleotide exchange factor recruited to beads by Ran-guanosine diphosphate (Ran GDP). Thus, concentration of Ran-GDP followed by generation of Ran-GTP is sufficient to induce NE assembly. PMID- 10827955 TI - Nipah virus: a recently emergent deadly paramyxovirus. AB - A paramyxovirus virus termed Nipah virus has been identified as the etiologic agent of an outbreak of severe encephalitis in people with close contact exposure to pigs in Malaysia and Singapore. The outbreak was first noted in late September 1998 and by mid-June 1999, more than 265 encephalitis cases, including 105 deaths, had been reported in Malaysia, and 11 cases of encephalitis or respiratory illness with one death had been reported in Singapore. Electron microscopic, serologic, and genetic studies indicate that this virus belongs to the family Paramyxoviridae and is most closely related to the recently discovered Hendra virus. We suggest that these two viruses are representative of a new genus within the family Paramyxoviridae. Like Hendra virus, Nipah virus is unusual among the paramyxoviruses in its ability to infect and cause potentially fatal disease in a number of host species, including humans. PMID- 10827956 TI - Granuloma-specific expression of Mycobacterium virulence proteins from the glycine-rich PE-PGRS family. AB - Pathogenic mycobacteria, including the agent of tuberculosis, Mycobacterium tuberculosis, must replicate in macrophages for long-term persistence within their niche during chronic infection: organized collections of macrophages and lymphocytes called granulomas. We identified several genes preferentially expressed when Mycobacterium marinum, the cause of fish and amphibian tuberculosis, resides in host granulomas and/or macrophages. Two were homologs of M. tuberculosis PE/PE-PGRS genes, a family encoding numerous repetitive glycine rich proteins of unknown function. Mutation of two PE-PGRS genes produced M. marinum strains incapable of replication in macrophages and with decreased persistence in granulomas. Our results establish a direct role in virulence for some PE-PGRS proteins. PMID- 10827957 TI - Regulation of B lymphocyte and macrophage development by graded expression of PU.1. AB - The ets family transcription factor PU.1 is required for the development of multiple lineages of the immune system. Using retroviral transduction of PU.1 complementary DNA into mutant hematopoietic progenitors, we demonstrate that differing concentrations of the protein regulate the development of B lymphocytes as compared with macrophages. A low concentration of PU. 1 protein induces the B cell fate, whereas a high concentration promotes macrophage differentiation and blocks B cell development. Conversely, a transcriptionally weakened mutant protein preferentially induces B cell generation. Our results suggest that graded expression of a transcription factor can be used to specify distinct cell fates in the hematopoietic system. PMID- 10827958 TI - Polyglutamine toxicity: a new idea for a tough problem. PMID- 10827959 TI - Effects of mobile buffers on facilitation: experimental and computational studies. AB - Facilitation is an important form of short-term plasticity that occurs in most synapses. At crayfish neuromuscular junctions, basal transmission and facilitation were significantly reduced after presynaptic introduction of "fast" high-affinity calcium buffers, and the decay of facilitation was accelerated. The existence of residual calcium during facilitation was also demonstrated. Computational modeling of three-dimensional buffered Ca(2+) diffusion and binding to secretory and facilitation targets suggest that the facilitation site is located away from a secretory trigger mediating exocytosis; otherwise, the facilitation site would be saturated by each action potential. Our simulations account for many characteristics of facilitation and effects of exogenous buffer, and suggest that facilitation is caused by residual calcium gaining access to a site distinct from the secretory trigger through restricted diffusion. PMID- 10827960 TI - On the temperature and pressure dependence of a range of properties of a type of water model commonly used in high-temperature protein unfolding simulations. AB - Molecular dynamics simulations of protein folding and unfolding are often carried out at temperatures (400-600 K) that are much higher than physiological or room temperature to speed up the (un)folding process. Use of such high temperatures changes both the protein and solvent properties considerably, compared to physiological or room temperature. Water models designed for use in conjunction with biomolecules, such as the simple point charge (SPC) model, have generally been calibrated at room temperature and pressure. To determine the distortive effect of high simulation temperatures on the behavior of such "room temperature" water models, the structural, dynamic, and thermodynamic properties of the much used SPC water model are investigated in the temperature range from 300 to 500 K. Both constant pressure and constant volume conditions, as used in protein simulations, were analyzed. We found that all properties analyzed change markedly with increasing temperature, but no phase transition in this temperature range was observed. PMID- 10827961 TI - Scroll wave dynamics in a three-dimensional cardiac tissue model: roles of restitution, thickness, and fiber rotation. AB - Scroll wave (vortex) breakup is hypothesized to underlie ventricular fibrillation, the leading cause of sudden cardiac death. We simulated scroll wave behaviors in a three-dimensional cardiac tissue model, using phase I of the Luo Rudy (LR1) action potential model. The effects of action potential duration (APD) restitution, tissue thickness, filament twist, and fiber rotation were studied. We found that APD restitution is the major determinant of scroll wave behavior and that instabilities arising from APD restitution are the main determinants of scroll wave breakup in this cardiac model. We did not see a "thickness-induced instability" in the LR1 model, but a minimum thickness is required for scroll breakup in the presence of fiber rotation. The major effect of fiber rotation is to maintain twist in a scroll wave, promoting filament bending and thus scroll breakup. In addition, fiber rotation induces curvature in the scroll wave, which weakens conduction and further facilitates wave break. PMID- 10827962 TI - Spatial buffering of potassium ions in brain extracellular space. AB - It has long been assumed that one important mechanism for the dissipation of local potassium gradients in the brain extracellular space is the so-called spatial buffer, generally associated with glial cells. To date, however, there has been no analytical description of the characteristic patterns of K(+) clearance mediated by such a mechanism. This study reanalyzed a mathematical model of Gardner-Medwin (1983, J. Physiol. (Lond.). 335:393-426) that had previously been solved numerically. Under suitable approximations, the transient solutions for the potassium concentrations and the corresponding membrane potentials of glial cells in a finite, parallel domain were derived. The analytic results were substantiated by numerical simulations of a detailed two-compartment model. This simulation explored the dependence of spatial buffer current and extracellular K(+) on the distribution of inward rectifier K(+) channels in the glial endfoot and nonendfoot membranes, the glial geometric length, and the effect of passive KCl uptake. Regarding the glial cells as an equivalent leaky cable, the analyses indicated that a maximum endfoot current occurs when the glial geometric length is equal to the corresponding electrotonic space constant. Consequently, a long glial process is unsuitable for spatial buffering, unless the axial space constant can match the length of the process. Finally, this study discussed whether the spatial buffer mechanism is able to efficiently transport K(+) over distances of more than several glial space constants. PMID- 10827963 TI - The mechanochemistry of V-ATPase proton pumps. AB - The vacuolar H(+)-ATPases (V-ATPases) are a universal class of proton pumps that are structurally similar to the F-ATPases. Both protein families are characterized by a membrane-bound segment (V(o), F(o)) responsible for the translocation of protons, and a soluble portion, (V(1), F(1)), which supplies the energy for translocation by hydrolyzing ATP. Here we present a mechanochemical model for the functioning of the V(o) ion pump that is consistent with the known structural features and biochemistry. The model reproduces a variety of experimental measurements of performance and provides a unified view of the many mechanisms of intracellular pH regulation. PMID- 10827965 TI - Analysis of the composite response of shear wave resonators to the attachment of mammalian cells. AB - The suitability of the quartz crystal microbalance (QCM) technique for monitoring the attachment and spreading of mammalian cells has recently been established. Different cell species were shown to generate an individual response of the QCM when they make contact with the resonator surface. Little is known, however, about the underlying mechanisms that determine the QCM signal for a particular cell type. Here we describe our results for different experimental approaches designed to probe the particular contributions of various subcellular compartments to the overall QCM signal. Using AC impedance analysis in a frequency range that closely embraces the resonators' fundamental frequency, we have explored the signal contribution of the extracellular matrix, the actin cytoskeleton, the medium that overlays the cell layer, as well as the liquid compartment that is known to exist between the basal plasma membrane and the culture substrate. Results indicate that the QCM technique is only sensitive to those parts of the cellular body that are involved in cell substrate adhesion and are therefore close to the resonator surface. Because of its noninvasive nature, sensitivity, and time resolution, the QCM is a powerful means of quantitatively studying various aspects of cell-substrate interactions. PMID- 10827964 TI - Cross-linking of cell surface receptors enhances cooperativity of molecular adhesion. AB - Cooperativity of molecular adhesion has been proposed as a mechanism for enhanced binding strength of adhesion molecules on the cell surface. Direct evidence for its mechanism, however, has been lacking until now. Atomic force microscopy (AFM) was used to measure the adhesive strength between concanavalin A (Con A) coupled to an AFM tip and Con A receptors on the surface of NIH3T3 fibroblast cells. Cross-linking of receptors with either glutaraldehyde or 3, 3'-dithio bis(sulfosuccinimidylproprionate) (DTSSP) led to an increase in adhesion that could be attributed to enhanced cooperativity among adhesion complexes. An increase in loading rate due to greater stiffness of fixed cells also contributed to the twofold increase in binding strength. These results show that receptor cross-linking can greatly contribute to a total increase in cell adhesion by creating a shift toward cooperative binding of receptors. PMID- 10827966 TI - Efficiency of platelet adhesion to fibrinogen depends on both cell activation and flow. AB - The kinetics of adhesion of platelets to fibrinogen (Fg) immobilized on polystyrene latex beads (Fg-beads) was determined in suspensions undergoing Couette flow at well-defined homogeneous shear rates. The efficiency of platelet adhesion to Fg-beads was compared for ADP-activated versus "resting" platelets. The effects of the shear rate (100-2000 s(-1)), Fg density on the beads (24-2882 Fg/microm(2)), the concentration of ADP used to activate the platelets, and the presence of soluble fibrinogen were assessed. "Resting" platelets did not specifically adhere to Fg-beads at levels detectable with our methodology. The apparent efficiency of platelet adhesion to Fg-beads readily correlated with the proportion of platelets "quantally" activated by doses of ADP, i.e., only ADP activated platelets appeared to adhere to Fg-beads, with a maximal adhesion efficiency of 6-10% at shear rates of 100-300 s(-1), decreasing with increasing shear rates up to 2000 s(-1). The adhesion efficiency was found to decrease by only threefold when decreasing the density of Fg at the surface of the beads by 100-fold, with only moderate decreases in the presence of physiologic concentrations of soluble Fg. These adhesive interactions were also compared using activated GPIIbIIIa-coated beads. Our studies provide novel model particles for studying platelet adhesion relevant to hemostasis and thrombosis, and show how the state of activation of the platelet and the local flow conditions regulate Fg-dependent adhesion. PMID- 10827967 TI - A membrane bending model of outer hair cell electromotility. AB - We propose a new mechanism for outer hair cell electromotility based on electrically induced localized changes in the curvature of the plasma membrane (flexoelectricity). Electromechanical coupling in the cell's lateral wall is modeled in terms of linear constitutive equations for a flexoelectric membrane and then extended to nonlinear coupling based on the Langevin function. The Langevin function, which describes the fraction of dipoles aligned with an applied electric field, is shown to be capable of predicting the electromotility voltage displacement function. We calculate the electrical and mechanical contributions to the force balance and show that the model is consistent with experimentally measured values for electromechanical properties. The model rationalizes several experimental observations associated with outer hair cell electromotility and provides for constant surface area of the plasma membrane. The model accounts for the isometric force generated by the cell and explains the observation that the disruption of spectrin by diamide reduces force generation in the cell. We discuss the relation of this mechanism to other proposed models of outer hair cell electromotility. Our analysis suggests that rotation of membrane dipoles and the accompanying mechanical deformation may be the molecular mechanism of electromotility. PMID- 10827968 TI - Role of actin cortex in the subplasmalemmal transport of secretory granules in PC 12 cells. AB - In neuroendocrine PC-12 cells, evanescent-field fluorescence microscopy was used to track motions of green fluorescent protein (GFP)-labeled actin or GFP-labeled secretory granules in a thin layer of cytoplasm where cells adhered to glass. The layer contained abundant filamentous actin (F-actin) locally condensed into stress fibers. More than 90% of the granules imaged lay within the F-actin layer. One-third of the granules did not move detectably, while two-thirds moved randomly; the average diffusion coefficient was 23 x 10(-4) microm(2)/s. A small minority (<3%) moved rapidly and in a directed fashion over distances more than a micron. Staining of F-actin suggests that such movement occurred along actin bundles. The seemingly random movement of most other granules was not due to diffusion since it was diminished by the myosin inhibitor butanedione monoxime, and blocked by chelating intracellular Mg(2+) and replacing ATP with AMP-PNP. Mobility was blocked also when F-actin was stabilized with phalloidin, and was diminished when the actin cortex was degraded with latrunculin B. We conclude that the movement of granules requires metabolic energy, and that it is mediated as well as limited by the actin cortex. Opposing actions of the actin cortex on mobility may explain why its degradation has variable effects on secretion. PMID- 10827969 TI - Neuronal death and perinatal lethality in voltage-gated sodium channel alpha(II) deficient mice. AB - Neural activity is crucial for cell survival and fine patterning of neuronal connectivity during neurodevelopment. To investigate the role in vivo of sodium channels (NaCh) in these processes, we generated knockout mice deficient in brain NaChalpha(II). NaChalpha(II)(-/-) mice were morphologically and organogenically indistinguishable from their NaChalpha(+/-) littermates. Notwithstanding, NaChalpha(II)(-/-) mice died perinatally with severe hypoxia and massive neuronal apoptosis, notably in the brainstem. Sodium channel currents recorded from cultured neurons of NaChalpha(II)(-/-) mice were sharply attenuated. Death appears to arise from severe hypoxia consequent to the brainstem deficiency of NaChalpha(II). NaChalpha(II) expression is, therefore, redundant for embryonic development but essential for postnatal survival. PMID- 10827970 TI - Poly-L-glutamine forms cation channels: relevance to the pathogenesis of the polyglutamine diseases. AB - We report that long-chain poly-L-glutamine forms cation-selective channels when incorporated into artificial planar lipid bilayer membranes. The channel was permeable to alkali cations and H(+) ions and virtually impermeable to anions; the selectivity sequence based on the single-channel conductance was H(+) >> Cs(+) > K(+) > Na(+). The cation channel was characterized by long-lived open states (often lasting for several minutes to tens of minutes) interrupted by brief closings. The appearance of the channel depended critically on the length of polyglutamine chains; ion channels were observed with 40-residue stretches, whereas no significant conductance changes were detected with 29-residue tracts. The channel-forming threshold length of poly-L-glutamine was thus between 29 and 40 residues. A molecular mechanics calculation suggests a mu-helix (. Biophys. J. 69:1130-1141) as a candidate molecular structure of the channel. The channel forming nature of long-chain poly-L-glutamine may provide a clue to the elucidation of the pathogenetic mechanism of the polyglutamine diseases, a group of inherited neurodegenerative disorders including Huntington's disease. PMID- 10827971 TI - Molecular dynamics of the KcsA K(+) channel in a bilayer membrane. AB - Molecular dynamics (MD) simulations of an atomic model of the KcsA K(+) channel embedded in an explicit dipalmitoylphosphatidylcholine (DPPC) phospholipid bilayer solvated by a 150 mM KCl aqueous salt solution are performed and analyzed. The model includes the KcsA K(+) channel, based on the recent crystallographic structure of, Science. 280:69-77), 112 DPPC, K(+) and Cl(-) ions, as well as over 6500 water molecules for a total of more than 40,000 atoms. Three K(+) ions are explicitly included in the pore. Two are positioned in the selectivity filter on the extracellular side and one in the large water-filled cavity. Different starting configurations of the ions and water molecules in the selectivity filter are considered, and MD trajectories are generated for more than 4 ns. The conformation of KcsA is very stable in all of the trajectories, with a global backbone root mean square (RMS) deviation of less than 1.9 A with respect to the crystallographic structure. The RMS atomic fluctuations of the residues surrounding the selectivity filter on the extracellular side of the channel are significantly lower than those on the intracellular side. The motion of the residues with aromatic side chains surrounding the selectivity filter (Trp(67), Trp(68), Tyr(78), and Tyr(82)) is anisotropic with the smallest RMS fluctuations in the direction parallel to the membrane plane. A concerted dynamic transition of the three K(+) ions in the pore is observed, during which the K(+) ion located initially in the cavity moves into the narrow part of the selectivity filter, while the other two K(+) ions move toward the extracellular side. A single water molecule is stabilized between each pair of ions during the transition, suggesting that each K(+) cation translocating through the narrow pore is accompanied by exactly one water molecule, in accord with streaming potential measurements (, Biophys. J. 55:367-371). The displacement of the ions is coupled with the structural fluctuations of Val(76) and Gly(77), in the selectivity filter, as well as the side chains of Glu(71), Asp(80), and Arg(89), near the extracellular side. Thus the mechanical response of the channel structure at distances as large as 10-20 A from the ions in the selectivity filter appears to play an important role in the concerted transition. PMID- 10827972 TI - GOLAC: an endogenous anion channel of the Golgi complex. AB - The Golgi complex is present in every eukaryotic cell and functions in posttranslational modifications and sorting of proteins and lipids to post-Golgi destinations. Both functions require an acidic lumenal pH and transport of substrates into and by-products out of the Golgi lumen. Endogenous ion channels are expected to be important for these features, but none has been described. Ion channels from an enriched Golgi fraction cleared of transiting proteins were incorporated into planar lipid bilayers. Eighty percent of the single-channel recordings revealed the same anion channel. This channel has novel properties and has been named GOLAC (Golgi anion channel). The channel has six subconductance states with a maximum conductance of 130 pS, is open over 95% of the time, and is not voltage-gated. Significant for Golgi function, the channel conductance is increased by reduction of pH on the lumenal surface. This channel may serve two nonexclusive functions: providing counterions for the acidification of the Golgi lumen by the H(+)-ATPase and removal of inorganic phosphate generated by glycosylation and sulfation of proteins and lipids in the Golgi. PMID- 10827973 TI - Homology modeling and molecular dynamics simulation studies of an inward rectifier potassium channel. AB - A homology model has been generated for the pore-forming domain of Kir6.2, a component of an ATP-sensitive K channel, based on the x-ray structure of the bacterial channel KcsA. Analysis of the lipid-exposed and pore-lining surfaces of the model reveals them to be compatible with the known features of membrane proteins and Kir channels, respectively. The Kir6.2 homology model was used as the starting point for nanosecond-duration molecular dynamics simulations in a solvated phospholipid bilayer. The overall drift from the model structure was comparable to that seen for KcsA in previous similar simulations. Preliminary analysis of the interactions of the Kir6.2 channel model with K(+) ions and water molecules during these simulations suggests that concerted single-file motion of K(+) ions and water through the selectivity filter occurs. This is similar to such motion observed in simulations of KcsA. This suggests that a single-filing mechanism is conserved between different K channel structures and may be robust to changes in simulation details. Comparison of Kir6.2 and KcsA suggests some degree of flexibility in the filter, thus complicating models of ion selectivity based upon a rigid filter. PMID- 10827974 TI - Rapid and slow voltage-dependent conformational changes in segment IVS6 of voltage-gated Na(+) channels. AB - Mutations in segment IVS6 of voltage-gated Na(+) channels affect fast inactivation, slow-inactivation, local anesthetic action, and batrachotoxin (BTX) action. To detect conformational changes associated with these processes, we substituted a cysteine for a valine at position 1583 in the rat adult skeletal muscle sodium channel alpha-subunit, and examined the accessibility of the substituted cysteine to modification by 2-aminoethyl methanethiosulfonate (MTS EA) in excised macropatches. MTS-EA causes an irreversible reduction in the peak current when applied both internally and externally, with a reaction rate that is strongly voltage-dependent. The rate increased when exposures to MTS-EA occurred during brief conditioning pulses to progressively more depolarized voltages, but decreased when exposures occurred at the end of prolonged depolarizations, revealing two conformational changes near site 1583, one coupled to fast inactivation, and one tightly associated with slow inactivation. Tetraethylammonium, a pore blocker, did not affect the reaction rate from either direction, while BTX, a lipophilic activator of sodium channels, completely prevented the modification reaction from occurring from either direction. We conclude that there are two inactivation-associated conformational changes in the vicinity of site 1583, that the reactive site most likely faces away from the pore, and that site 1583 comprises part of the BTX receptor. PMID- 10827975 TI - Expression of the alpha(2)delta subunit interferes with prepulse facilitation in cardiac L-type calcium channels. AB - We investigated the role of the accessory alpha(2)delta subunit on the voltage dependent facilitation of cardiac L-type Ca(2+) channels (alpha(1C)). alpha(1C) Channels were coexpressed in Xenopus oocytes with beta(3) and alpha(2)delta calcium channel subunits. In alpha(1C) + beta(3), the amplitude of the ionic current (measured during pulses to 10 mV) was in average approximately 1.9-fold larger after the application of a 200-ms prepulse to +80 mV. This phenomenon, commonly referred to as voltage-dependent facilitation, was not observed when alpha(2)delta was coexpressed with alpha(1C) + beta(3). In alpha(1C) + beta(3), the prepulse produced a left shift ( approximately 40 mV) of the activation curve. Instead, the activation curve for alpha(1C) + beta(3) + alpha(2)delta was minimally affected by the prepulse and had a voltage dependence very similar to the G-V curve of the alpha(1C) + beta(3) channel facilitated by the prepulse. Coexpression of alpha(2)delta with alpha(1C) + beta(3) seems to mimic the prepulse effect by shifting the activation curve toward more negative potentials, leaving little room for facilitation. The facilitation of alpha(1C) + beta(3) was associated with an increase of the charge movement. In the presence of alpha(2)delta, the charge remained unaffected after the prepulse. Coexpression of alpha(2)delta seems to set all the channels in a conformational state from where the open state can be easily reached, even without prepulse. PMID- 10827977 TI - Resolution of patch capacitance recordings and of fusion pore conductances in small vesicles. AB - We investigated the noise levels in cell-attached patch capacitance recordings with a lock-in amplifier. The capacitance noise level decreases with increasing sine wave frequency up to 20-40 kHz. With a 20-mV rms sine wave the rms noise level above 8 kHz is <50 aF. With increasing sine wave amplitudes a further reduction down to 14 aF could be achieved. Capacitance measurements with a lock in amplifier may also be used to measure the conductance of fusion pores connecting the vesicular lumen to the extracellular space. It is estimated that at noise levels of 14 aF fusion pore conductances between 20 pS and 700 pS may be resolved in vesicles with 380-aF capacitance by using a 50-kHz sine wave. This corresponds to vesicles with a approximately 110-nm diameter. It is suggested that with low-noise techniques fusion pores may be detectable in vesicles approaching the size of large synaptic vesicles. PMID- 10827976 TI - Molecular determinants of anion selectivity in the cystic fibrosis transmembrane conductance regulator chloride channel pore. AB - Ionic selectivity in many cation channels is achieved over a short region of the pore known as the selectivity filter, the molecular determinants of which have been identified in Ca(2+), Na(+), and K(+) channels. However, a filter controlling selectivity among different anions has not previously been identified in any Cl(-) channel. In fact, because Cl(-) channels are only weakly selective among small anions, and because their selectivity has proved so resistant to site directed mutagenesis, the very existence of a discrete anion selectivity filter has been called into question. Here we show that mutation of a putative pore lining phenylalanine residue, F337, in the sixth membrane-spanning region of the cystic fibrosis transmembrane conductance regulator (CFTR) Cl(-) channel, dramatically alters the relative permeabilities of different anions in the channel. Specifically, mutations that reduce the size of the amino acid side chain present at this position virtually abolish the relationship between anion permeability and hydration energy, a relationship that characterizes the anion selectivity not only of wild-type CFTR, but of most classes of Cl(-) channels. These results suggest that the pore of CFTR may indeed contain a specialized region, analogous to the selectivity filter of cation channels, at which discrimination between different permeant anions takes place. Because F337 is adjacent to another amino acid residue, T338, which also affects anion selectivity in CFTR, we suggest that selectivity is predominantly determined over a physically discrete region of the pore located near these important residues. PMID- 10827978 TI - Excitability of the soma in central nervous system neurons. AB - The ability of the soma of a spinal dorsal horn neuron, a spinal ventral horn neuron (presumably a motoneuron), and a hippocampal pyramidal neuron to generate action potentials was studied using patch-clamp recordings from rat spinal cord slices, the "entire soma isolation" method, and computer simulations. By comparing original recordings from an isolated soma of a dorsal horn neuron with simulated responses, it was shown that computer models can be adequate for the study of somatic excitability. The modeled somata of both spinal neurons were unable to generate action potentials, showing only passive and local responses to current injections. A four- to eightfold increase in the original density of Na(+) channels was necessary to make the modeled somata of both spinal neurons excitable. In contrast to spinal neurons, the modeled soma of the hippocampal pyramidal neuron generated spikes with an overshoot of +9 mV. It is concluded that the somata of spinal neurons cannot generate action potentials and seem to resist their propagation from the axon to dendrites. In contrast, the soma of the hippocampal pyramidal neuron is able to generate spikes. It cannot initiate action potentials in the intact neurons, but it can support their back propagation from the axon initial segment to dendrites. PMID- 10827979 TI - Strength of Ca(2+) binding to retinal lipid membranes: consequences for lipid organization. AB - There is evidence that membranes of rod outer segment (ROS) disks are a high affinity Ca(2+) binding site. We were interested to see if the high occurrence of sixfold unsaturated docosahexaenoic acid in ROS lipids influences Ca(2+)-membrane interaction. Ca(2+) binding to polyunsaturated model membranes that mimic the lipid composition of ROS was studied by microelectrophoresis and (2)H NMR. Ca(2+) association constants of polyunsaturated membranes were found to be a factor of approximately 2 smaller than constants of monounsaturated membranes. Furthermore, strength of Ca(2+) binding to monounsaturated membranes increased with the addition of cholesterol, while binding to polyunsaturated lipids was unaffected. The data suggest that the lipid phosphate groups of phosphatidylcholine (PC), phosphatidylethanolamine (PE), and phosphatidylserine (PS) in PC/PE/PS (4:4:1, mol/mol) are primary targets for Ca(2+). Negatively charged serine in PS controls Ca (2+) binding by lowering the electric surface potential and elevating cation concentration at the membrane/water interface. The influence of hydrocarbon chain unsaturation on Ca(2+) binding is secondary compared to membrane PS content. Order parameter analysis of individual lipids in the mixture revealed that Ca(2+) ions did not trigger lateral phase separation of lipid species as long as all lipids remained liquid-crystalline. However, depending on temperature and hydrocarbon chain unsaturation, the lipid with the highest chain melting temperature converted to the gel state, as observed for the monounsaturated phosphatidylethanolamine (PE) in PC/PE/PS (4:4:1, mol/mol) at 25 degrees C. PMID- 10827980 TI - Partitioning of amphiphiles between coexisting ordered and disordered phases in two-phase lipid bilayer membranes. AB - The partition coefficients (K(P)) of a series of single-chain and double-chain fluorescent amphiphiles, between solid ordered (P(beta') and L(beta)) and liquid disordered (L(alpha) of the type l(d)) lipid phases coexisting in the same lipid bilayer, was studied using steady-state fluorescence emission anisotropy. The single-chain amphiphiles were N-(7-nitrobenzoxa-2, 3-diazol-4-yl)-alkylamines, and the double-chain amphiphiles were N-(7-nitrobenzoxa-2, 3-diazol-4-yl) phosphatidylethanolamines with chain lengths of 12-18 carbon atoms. Saturated 18 carbon alkyl/acyl chain compounds were also compared with Delta(9)-cis unsaturated chains of the same chain length. The fluorescence anisotropy of the probes was examined in lipid bilayers (multilamellar vesicles) prepared from an equimolar mixture of dilauroylphosphatidylcholine and distearoylphosphatidylcholine and studied as a function of temperature through the entire temperature range of coexistence of ordered gel phases and a disordered fluid phase in this system. The unsaturated chain amphiphiles partitioned exclusively into the fluid phase whenever this phase was present, as did the saturated chain amphiphiles with the shortest chains (C(12:0)), while K(P) ranges between 1 and 2, in favor of the L(beta) solid phase, for the amphiphiles with long saturated (C(18:0)) alkyl/acyl chains, with intermediate behavior for the intermediate chain lengths. All probes appeared to be totally excluded from P(beta') solid (gel) phases. The technique was also used to determine partitioning of some of the probes between coexisting liquid ordered (cholesterol-containing) (l(o)) and liquid disordered (l(d)) L(alpha) phases. In this case the ratio of signal amplitude to noise allowed us to obtain a qualitative, but not quantitative, measure of the phase partitioning of the probes. We conclude that the partitioning behavior of the probes examined between coexisting l(o) and l(d) phases is qualitatively similar to that observed between solid ordered and liquid disordered phases. PMID- 10827981 TI - Asymmetrical ion-channel model inferred from two-dimensional crystallization of a peptide antibiotic. AB - The structural organization of ion channels formed in lipid membranes by amphiphilic alpha-helical peptides is deduced by applying direct structural methods to different lipid/alamethicin systems. Alamethicin represents a hydrophobic alpha-helical peptide antibiotic forming voltage-gated ion channels in lipid membranes. Here the first direct evidence for the existence of large scale two-dimensional crystalline domains of alamethicin helices, oriented parallel to the air/water interface, is presented using synchrotron x-ray diffraction, fluorescence microscopy, and surface pressure/area isotherms. Proofs are obtained that the antibiotic peptide injected into the aqueous phase under phospholipid monolayers penetrates these monolayers, phase separates, and forms domains within the lipid environment, keeping the same, parallel orientation of the alpha-helices with respect to the phospholipid/water interface. A new asymmetrical, "lipid-covered ring" model of the voltage-gated ion channel of alamethicin is inferred from the structural results presented, and the mechanism of ion-channel formation is discussed. PMID- 10827982 TI - Endocytosis switch controlled by transmembrane osmotic pressure and phospholipid number asymmetry. AB - The dynamics of endocytosis in living K562 cells was investigated after the osmotic pressure of the external medium was decreased and the transmembrane phospholipid number asymmetry was increased. When the external pressure was decreased by a factor of 0.54, a sudden inhibition of endocytosis was observed. Under these conditions, the endocytosis suddenly recovered after the phospholipid number asymmetry was increased. The phospholipid asymmetry was generated by the addition of exogenous phosphatidylserine, which is translocated by the endogenous flippase activity to the inner layer of the membrane. The recovery of endocytosis is thus consistent with the view that the phospholipid number asymmetry can act as a budding force for endocytosis. Moreover, we quantitatively predict both the inhibition and recovery of endocytosis as first-order phase transitions, using a general model that assumes the existence of a transmembrane surface tension asymmetry as the budding driving force. In this model, the tension asymmetry is considered to be elastically generated by the activity of phospholipid pumping. We finally propose that cells may trigger genetic transcription responses after the internalization of cytokine-receptor complexes, which could be controlled by variations in the cytosolic or external pressure. PMID- 10827983 TI - Comparative single-molecule and ensemble myosin enzymology: sulfoindocyanine ATP and ADP derivatives. AB - Single-molecule and macroscopic reactions of fluorescent nucleotides with myosin have been compared. The single-molecule studies serve as paradigms for enzyme catalyzed reactions and ligand-receptor interactions analyzed as individual stochastic processes. Fluorescent nucleotides, called Cy3-EDA-ATP and Cy5-EDA ATP, were derived by coupling the dyes Cy3.29.OH and Cy5.29.OH (compounds XI and XIV, respectively, in, Bioconjug. Chem. 4:105-111)) with 2'(3')-O-[N-(2 aminoethyl)carbamoyl]ATP (EDA-ATP). The ATP(ADP) analogs were separated into their respective 2'- and 3'-O-isomers, the interconversion rate of which was 30[OH(-)] s(-1) (0.016 h(-1) at pH 7.1) at 22 degrees C. Macroscopic studies showed that 2'(3')-O-substituted nucleotides had properties similar to those of ATP and ADP in their interactions with myosin, actomyosin, and muscle fibers, although the ATP analogs did not relax muscle as well as ATP did. Significant differences in the fluorescence intensity of Cy3-nucleotide 2'- and 3'-O-isomers in free solution and when they interacted with myosin were evident. Single molecule studies using total internal reflection fluorescence microscopy showed that reciprocal mean lifetimes of the nucleotide analogs interacting with myosin filaments were one- to severalfold greater than predicted from macroscopic data. Kinetic and equilibrium data of nucleotide-(acto)myosin interactions derived from single-molecule microscopy now have a biochemical and physiological framework. This is important for single-molecule mechanical studies of motor proteins. PMID- 10827984 TI - Actin and temperature effects on the cross-linking of the SH1-SH2 helix in myosin subfragment 1. AB - Past biochemical work on myosin subfragment 1 (S1) has shown that the bent alpha helix containing the reactive thiols SH1 (Cys(707)) and SH2 (Cys(697)) changes upon nucleotide and actin binding. In this study, we investigated the conformational dynamics of the SH1-SH2 helix in two actin-bound states of myosin and examined the effect of temperature on this helix, using five cross-linking reagents that are 5-15 A in length. Actin inhibited the cross-linking of SH1 to SH2 on both S1 and S1.MgADP for all of the reagents. Because the rate of SH2 modification was not altered by actin, the inhibition of cross-linking must result from a strong stabilization of the SH1-SH2 helix in the actin-bound states of S1. The dynamics of the helix is also influenced by temperature. At 25 degrees C, the rate constants for cross-linking in S1 alone are low, with values of approximately 0.010 min(-1) for all of the reagents. At 4 degrees C, the rate constants, except for the shortest reagent, range between 0.030 and 0.070 min( 1). The rate constants for SH2 modification in SH1-modified S1 show the opposite trend; they increase with the increases in temperature. The greater cross-linking at the lower temperature indicates destabilization of the SH1-SH2 helix at 4 degrees C. These results are discussed in terms of conformational dynamics of the SH1-SH2 helix. PMID- 10827985 TI - The effect of inorganic phosphate on force generation in single myofibrils from rabbit skeletal muscle. AB - In striated muscle, force generation and phosphate (P(i)) release are closely related. Alterations in the [P(i)] bathing skinned fibers have been used to probe key transitions of the mechanochemical coupling. Accuracy in this kind of studies is reduced, however, by diffusional barriers. A new perfusion technique is used to study the effect of [P(i)] in single or very thin bundles (1-3 microM in diameter; 5 degrees C) of rabbit psoas myofibrils. With this technique, it is possible to rapidly jump [P(i)] during contraction and observe the transient and steady-state effects on force of both an increase and a decrease in [P(i)]. Steady-state isometric force decreases linearly with an increase in log[P(i)] in the range 500 microM to 10 mM (slope -0.4/decade). Between 5 and 200 microM P(i), the slope of the relation is smaller ( approximately -0.07/decade). The rate constant of force development (k(TR)) increases with an increase in [P(i)] over the same concentration range. After rapid jumps in [P(i)], the kinetics of both the force decrease with an increase in [P(i)] (k(Pi(+))) and the force increase with a decrease in [P(i)] (k(Pi(-))) were measured. As observed in skinned fibers with caged P(i), k(Pi(+)) is about three to four times higher than k(TR), strongly dependent on final [P(i)], and scarcely modulated by the activation level. Unexpectedly, the kinetics of force increase after jumps from high to low [P(i)] is slower: k(Pi(-)) is indistinguishable from k(TR) measured at the same [P(i)] and has the same calcium sensitivity. PMID- 10827987 TI - Influence of ADP on cross-bridge-dependent activation of myofibrillar thin filaments. AB - Contraction of skeletal muscle is regulated by calcium at the level of the thin filament via troponin and tropomyosin. Studies have indicated that strong cross bridge binding is also involved in activation of the thin filament. To further test this, myofibrils were incubated with a wide range of fluorescent myosin subfragment 1(fS1) at pCa 9 or pCa 4 with or without ADP. Sarcomere fluorescence intensity and the fluorescence intensity ratio (non-overlap region/overlap region) were measured to determine the amount and location of bound fS1 in the myofibril. There was lower sarcomere fluorescence intensity with ADP compared to without ADP for both calcium levels. Similar data were obtained from biochemical measures of bound fS1, validating the fluorescence microscopy measurements. The intensity ratio, which is related to activation of the thin filament, increased with increasing [fS1] with or without ADP. At pCa 9, the fluorescence intensity ratio was constant until 80-160 nM fS1 without ADP conditions, then it went up dramatically and finally attained saturation. The dramatic shift of the ratio demonstrated the cooperative character of strong cross-bridge binding, and this was not observed at high calcium. A similar pattern was observed with ADP in that the ratio was right-shifted with respect to total [fS1]. Saturation was obtained with both the fluorescence intensity and ratio data. Plots of intensity ratio as a function of normalized sarcomere intensity (bound fS1) showed little difference between with and without ADP. This suggests that the amount of strongly bound fS1, not fS1 state (with or without ADP) is related to activation of the thin filament. PMID- 10827986 TI - Interaction of myosin with F-actin: time-dependent changes at the interface are not slow. AB - The kinetics of formation of the actin-myosin complex have been reinvestigated on the minute and second time scales in sedimentation and chemical cross-linking experiments. With the sedimentation method, we found that the binding of the skeletal muscle myosin motor domain (S1) to actin filament always saturates at one S1 bound to one actin monomer (or two S1 per actin dimer), whether S1 was added slowly (17 min between additions) or rapidly (10 s between additions) to an excess of F-actin. The carbodiimide (1-ethyl-3-(3-dimethylaminopropyl) carbodiimide, EDC)-induced cross-linking of the actin-S1 complex was performed on the subsecond time scale by a new approach that combines a two-step cross-linking protocol with the rapid flow-quench technique. The results showed that the time courses of S1 cross-linking to either of the two actin monomers are identical: they are not dependent on the actin/S1 ratio in the 0.3-20-s time range. The overall data rule out a mechanism by which myosin rolls from one to the other actin monomer on the second or minute time scales. Rather, they suggest that more subtle changes occur at the actomyosin interface during the ATP cycle. PMID- 10827988 TI - Temperature change does not affect force between single actin filaments and HMM from rabbit muscles. AB - The temperature dependence of sliding force, velocity, and unbinding force was studied on actin filaments when they were placed on heavy meromyosin (HMM) attached to a glass surface. A fluorescently labeled actin filament was attached to the gelsolin-coated surface of a 1-microm polystyrene bead. The bead was trapped by optical tweezers, and HMM-actin interaction was performed at 20-35 degrees C to examine whether force is altered by the temperature change. Our experiments demonstrate that sliding force increased moderately with temperature (Q(10) = 1.6 +/- 0.2, +/-SEM, n = 9), whereas the velocity increased significantly (Q(10) = 2.9 +/- 0.4, n = 10). The moderate increase in force is caused by the increased number of available cross-bridges for actin interaction, because the cross-bridge number similarly increased with temperature (Q(10) = 1. 5 +/- 0.2, n = 3) when measured during rigor induction. We further found that unbinding force measured during the rigor condition did not differ with temperature. These results indicate that the amount of force each cross-bridge generates is fixed, and it does not change with temperature. We found that the above generalization was not modified in the presence of 1 mM MgADP or 8 mM phosphate. PMID- 10827989 TI - Different myofilament nearest-neighbor interactions have distinctive effects on contractile behavior. AB - Cooperativity in contractile behavior of myofilament systems almost assuredly arises because of interactions between neighboring sites. These interactions may be of different kinds. Tropomyosin thin-filament regulatory units may have neighbors in steric blocking positions (off) or steric permissive positions (on). The position of these neighbors influence the tendency for the regulatory unit to assume the on or off state. Likewise, the tendency of a myosin cross-bridge to achieve a force-bearing state may be influenced by whether neighboring cross bridges are in force-bearing states. Also, a cross-bridge in the force-bearing state may influence the tendency of a regulatory unit to enter the on state. We used a mathematical model to examine the influence of each of these three kinds of neighbor interactions on the steady-state force-pCa relation and on the dynamic force redevelopment process. Each neighbor interaction was unique in its effects on maximal Ca(2+)-activated force, position, and symmetry of the force pCa curve and on the Hill coefficient. Also, each neighbor interaction had a distinctive effect on the time course of force development as assessed by its rate coefficient, k(dev). These diverse effects suggest that variations in all three kinds of nearest-neighbor interactions may be responsible for a wide variety of currently unexplained observations of myofilament contractile behavior. PMID- 10827990 TI - Influence of ionic strength on the actomyosin reaction steps in contracting skeletal muscle fibers. AB - Muscle contraction occurs as the result of actin-myosin interaction, which is mediated by the intermolecular forces exerted at the actin-myosin interface. To obtain information about the nature of these intermolecular forces, we tested the sensitivity of various contractile parameters of skinned skeletal muscle fibers to ionic strength (IS) at 3-5 degrees C; IS variation is a useful technique for distinguishing between ionic and nonionic (primarily hydrophobic) types of intermolecular forces. The most striking effect of elevated IS was the strong suppression of isometric tension. However, none of the measured parameters suggested a corresponding decrease in the number of force-generating myosin heads on actin. The rate of actin-myosin association seemed to be only modestly IS sensitive. The following force-generating isomerization was apparently IS insensitive. The dissociation of the force-generating actomyosin complex was decelerated by elevated IS, contrary to the expectation from the suppressed isometric tension. These results led us to conclude that an IS-sensitive step, responsible for the large suppression of tension, occurs after force-generating isomerization but before dissociation. The present study suggests that the actomyosin interaction is generally nonionic in nature, but there are at least two ionic processes, one at the beginning and the other close to the end of the actomyosin interaction. PMID- 10827991 TI - The M intermediate of Pharaonis phoborhodopsin is photoactive. AB - The retinal protein phoborhodopsin (pR) (also called sensory rhodopsin II) is a specialized photoreceptor pigment used for negative phototaxis in halobacteria. Upon absorption of light, the pigment is transformed into a short-wavelength intermediate, M, that most likely is the signaling state (or its precursor) that triggers the motility response of the cell. The M intermediate thermally decays into the initial pigment, completing the cycle of transformations. In this study we attempted to determine whether M can be converted into the initial state by light. The M intermediate was trapped by the illumination of a water glycerol suspension of phoborhodopsin from Natronobacterium pharaonis called pharaonis phoborhodopsin (ppR) with yellow light (>450 nm) at -50 degrees C. The M intermediate absorbing at 390 nm is stable in the dark at this temperature. We found, however, that M is converted into the initial (or spectrally similar) state with an absorption maximum at 501 nm upon illumination with 380-nm light at -60 degrees C. The reversible transformations ppR if M are accompanied by the perturbation of tryptophan(s) and probably tyrosine(s) residues, as reflected by changes in the UV absorption band. Illumination at lower temperature (-160 degrees C) reveals two intermediates in the photoconversion of M, which we termed M' (or M'(404)) and ppR' (or ppR'(496)). A third photoproduct, ppR'(504), is formed at -110 degrees C during thermal transformations of M'(404) and ppR'(496). The absorption spectrum of M'(404) (maximum at 404 nm) consists of distinct vibronic bands at 362, 382, 404, and 420 nm that are different from the vibronic bands of M at 348, 368, 390, and 415 nm. ppR'(496) has an absorption band that is shifted to shorter wavelengths by 5 nm compared to the initial ppR, whereas ppR'(504) is redshifted by at least 3 nm. As in bacteriorhodopsin, photoexcitation of the M intermediate of ppR and, presumably, photoisomerization of the chromophore during the M --> M' transition result in a dramatic increase in the proton affinity of the Schiff base, followed by its reprotonation during the M' --> ppR' transition. Because the latter reaction occurs at very low temperature, the proton is most likely taken from the counterion (Asp(75)) rather than from the bulk. The phototransformation of M reveals a certain heterogeneity of the pigment, which probably reflects different populations of M or its photoproduct M'. Photoconversion of the M intermediate provides a possible pathway for photoreception in halobacteria and a useful tool for studying the mechanisms of signal transduction by phoborhodopsin (sensory rhodopsin II). PMID- 10827992 TI - The bound electron acceptors in green sulfur bacteria: resolution of the g-tensor for the F(X) iron-sulfur cluster in Chlorobium tepidum. AB - The photosynthetic reaction center (RC) of green sulfur bacteria contains two [4Fe-4S] clusters named F(A) and F(B), by analogy with photosystem I (PS I). PS I also contains an interpolypeptide [4Fe-4S] cluster named F(X); however, spectroscopic evidence for an analogous iron-sulfur cluster in green sulfur bacteria remains equivocal. To minimize oxidative damage to the iron-sulfur clusters, we studied the sensitivity of F(A) and F(B) to molecular oxygen in whole cells of Chlorobium vibrioforme and Chlorobium tepidum and obtained highly photoactive membranes and RCs from Cb. tepidum by adjusting isolation conditions to maximize the amplitude of the F(A)(-)/F(B)(-) electron paramagnetic resonance signal at g = 1.89 (measured at 126 mW of microwave power and 14 K) relative to the P840(+) signal at g = 2.0028 (measured at 800 microW of microwave power and 14 K). In these optimized preparations we were able to differentiate F(X)(-) from F(A)(-)/F(B)(-) by their different relaxation properties. At temperatures between 4 and 9 K, isolated membranes and RCs of Cb. tepidum show a broad peak at g = 2.12 and a prominent high-field trough at g = 1.76 (measured at 126 mW of microwave power). The complete g-tensor of F(X)(-), extracted by numerical simulation, yields principal values of 2.17, 1.92, and 1. 77 and is similar to F(X) in PS I. An important difference from PS I is that because the bound cytochrome is available as a fast electron donor in Chlorobium, it is not necessary to prereduce F(A) and F(B) to photoaccumulate F(X)(-). PMID- 10827993 TI - Buffer effects on electric signals of light-excited bacteriorhodopsin. AB - Buffers change the electric signals of light-excited bacteriorhodopsin molecules in purple membrane if their concentration and the pH of the low-salt solution are properly selected. "Positive" buffers produce a positive component, and "negative" buffers a negative component in addition to the signals due to proton pumping. Measurement of the buffer effects in the presence of glycyl-glycine or bis-tris propane revealed an increase of approximately 2 and a change of sign and a decrease to approximately -0.5 in the translocated charge in these cases, respectively. These factors do not depend on temperature. The Arrhenius parameters established from the evaluation of the kinetics indicate activation enthalpies of 35-40 kJ/mol and negative activation entropies for the additional signals. These values agree with those found by surface-bound pH-sensitive probes in the search of the timing of proton release and uptake. The electric signals were also measured in the case of D(2)O solutions with similar results, except for the increased lifetimes. We offer a unified explanation for the data obtained with surface-bound probes and electric signals based on the clusters at extracellular and cytoplasmic sites of bacteriorhodopsin participating in proton release and uptake. PMID- 10827994 TI - Characterization of conditions required for X-Ray diffraction experiments with protein microcrystals. AB - The x-ray exposure at which significant radiation damage occurs has been quantified for frozen crystals of bacteriorhodopsin. The maximum exposure to approximately 11-keV x-rays that can be tolerated for high-resolution diffraction experiments is found to be approximately 10(10) photons/microm(2), very close to the value predicted from limits that were measured earlier for electron diffraction exposures. Sample heating, which would further reduce the x-ray exposure that could be tolerated, is not expected to be significant unless the x ray flux density is well above 10(9) photons/s-microm(2). Crystals of bacteriorhodopsin that contain approximately 10(11) unit cells are found to be large enough to give approximately 100 high-resolution diffraction patterns, each covering one degree of rotation. These measurements are used to develop simple rules of thumb for the minimum crystal size that can be used to record x-ray diffraction data from protein microcrystals. For work with very small microcrystals to be realized in practice, however, it is desirable that there be a significant reduction in the level of background scattering. Background reduction can readily be achieved by improved microcollimation of the x-ray beam, and additional gains can be realized by the use of helium rather than nitrogen in the cold gas stream that is used to keep the protein crystals frozen. PMID- 10827995 TI - Two-dimensional crystallization on lipid monolayers and three-dimensional structure of sticholysin II, a cytolysin from the sea anemone Stichodactyla helianthus. AB - Sticholysin II (Stn II), a potent cytolytic protein isolated from the sea anemone Stichodactyla helianthus, has been crystallized on lipid monolayers. With Fourier based methods, a three-dimensional (3D) model of Stn II, up to a resolution of 15 A, has been determined. The two-sided plane group is p22(1)2, with dimensions a = 98 A, b = 196 A. The 3D model of Stn II displays a Y-shaped structure, slightly flattened, with a small curvature along its longest dimension (51 A). This protein, with a molecular mass of 19. 2 kDa, is one of the smallest structures reconstructed with this methodology. Two-dimensional (2D) crystals of Stn II on phosphatidylcholine monolayers present a unit cell with two tetrameric motifs, with the monomers in two different orientations: one with its longest dimension lying on the crystal plane and the other with this same axis leaning at an angle of approximately 60 degrees with the crystal plane. PMID- 10827997 TI - Thermal denaturing of bacteriorhodopsin by X-Ray scattering from oriented purple membranes. AB - We present a temperature-dependent x-ray diffraction study of thin films of purple membranes (PMs) with the native membrane protein bacteriorhodopsin (BR). The high degree of alignment with respect to the silicon substrates allows for the application of modern interface-sensitive scattering techniques. Here we focus on the structural changes of BR in PMs at the thermal denaturing transition. A partial unfolding of the helices is observed rather than the complete unfolding process known from helix to coil transitions. While BR remains threaded into the lipid bilayer in the denatured state, changes in the short range lateral structures are associated with the partial unfolding of the transmembrane helices. PMID- 10827996 TI - Comparison of the solution conformation and dynamics of antifreeze glycoproteins from Antarctic fish. AB - The (1)H- and (13)C-NMR spectra of antifreeze glycoprotein fractions 1-5 from Antarctic cod have been assigned, and the dynamics have been measured using (13)C relaxation at two temperatures. The chemical shifts and absence of non-sequential (1)H-(1)H NOEs are inconsistent with a folded, compact structure. (13)C relaxation measurements show that the protein has no significant long-range order, and that the local correlation times are adequately described by a random coil model. Hydroxyl protons of the sugar residues were observed at low temperature, and the presence of exchange-mediated ROEs to the sugar indicate extensive hydration. The conformational properties of AFGP1-5 are compared with those of the previously examined 14-mer analog AFGP8, which contains proline residues in place of some alanine residues (Lane, A. N., L. M. Hays, R. E. Feeney, L. M. Crowe, and J. H. Crowe. 1998. Protein Sci. 7:1555-1563). The infrared (IR) spectra of AFGP8 and AFGP1-5 in the amide I region are quite different. The presence of a wide distribution of backbone torsion angles in AFGP1-5 leads to a rich spectrum of frequencies in the IR spectrum, as interconversion among conformational states is slow on the IR frequency time scale. However, these transitions are fast on the NMR chemical shift time scales. The restricted motions for AFGP8 may imply a narrower distribution of possible o, psi angles, as is observed in the IR spectrum. This has significance for attempts to quantify secondary structures of proteins by IR in the presence of extensive loops. PMID- 10827998 TI - Solubility of fluoromethemoglobin S: effect of phosphate and temperature on polymerization. AB - The polymerization properties of the fully liganded fluoromet derivative of hemoglobin S (FmetHb S) were investigated by electron microscopy and absorption spectroscopy. Polymerization progress curves, as measured by increasing sample turbidity at 700 nm, exhibit a delay time (t(d)) consistent with the double nucleation mechanism. The pattern of fiber growth, as monitored by electron microscopy, is also indicative of a heterogeneous nucleation process, and dimensions of the fibers were found to be comparable to that of deoxyHb S. The polymerization rate constant (1/t(d)) depends exponentially on Hb S concentration, and the size of the homogeneous and heterogeneous nuclei also depend on FmetHb S concentration. As for deoxyHb S, higher concentrations of protein and phosphate favor fiber formation, while lower temperatures inhibit polymerization. Solubility experiments reveal, however, that eight times more FmetHb S is required for polymerization. The current studies further show that reaction order is independent of phosphate concentration if Hb S activity and not concentration is considered. The allosteric effector, inositol hexaphosphate (IHP), promotes fiber formation, and temperature-dependent reaggregation of FmetHb S suggests that IHP stabilizes pregelation aggregates. These studies show that FmetHb S resembles deoxyHb S in many of its polymerization properties; however, IHP-bound FmetHb S potentially provides a unique avenue for future studies of the early stages of Hb S polymerization and the effect of tertiary and quaternary protein structure on the polymerization process. PMID- 10827999 TI - Multiple geminate ligand recombinations in human hemoglobin. AB - The geminate ligand recombination reactions of photolyzed carbonmonoxyhemoglobin were studied in a nanosecond double-excitation-pulse time-resolved absorption experiment. The second laser pulse, delayed by intervals as long as 400 ns after the first, provided a measure of the geminate kinetics by rephotolyzing ligands that have recombined during the delay time. The peak-to-trough magnitude of the Soret band photolysis difference spectrum measured as a function of the delay between excitation pulses showed that the room temperature kinetics of geminate recombination in adult human hemoglobin are best described by two exponential processes, with lifetimes of 36 and 162 ns. The relative amounts of bimolecular recombination to T- and R-state hemoglobins and the temperature dependence of the submicrosecond kinetics between 283 and 323 K are also consistent with biexponential kinetics for geminate recombination. These results are discussed in terms of two models: geminate recombination kinetics modulated by concurrent protein relaxation and heterogeneous kinetics arising from alpha and beta chain differences. PMID- 10828001 TI - Visualization of myoglobin-facilitated mitochondrial O(2) delivery in a single isolated cardiomyocyte. AB - The purpose of the present study was to visualize myoglobin-facilitated oxygen delivery to mitochondria at a critical mitochondrial oxygen supply in single isolated cardiomyocytes of rats. Using the autofluorescence of mitochondrial reduced nicotinamide adenine dinucleotide (phosphate) (NAD(P)H), the mitochondrial oxygen supply was imaged from approximately 1.4 microm inside the cell surface at a subcellular spatial resolution. Significant radial gradients of intracellular oxygenation were produced by superfusing the cell suspension with a mixed gas containing 2-4% oxygen while stimulating mitochondrial respiration with an uncoupler of oxidative phosphorylation. Augmentation of the NAD(P)H fluorescence started from the core of the cell (anoxic core) and progressively expanded toward the plasma membrane, as the extracellular Po(2) was lowered. Inactivation of cytosolic myoglobin by 5 mM NaNO(2) significantly enlarged such anoxic regions. Nitrite affected neither mitochondrial respiration in uncoupled cells nor the relationship between Po(2) and the NAD(P)H fluorescence in coupled cells. Thus we conclude that myoglobin significantly facilitates intracellular oxygen transport at a critical level of mitochondrial oxygen supply in single cardiomyocytes. PMID- 10828000 TI - Ligand-induced conformational changes in tissue transglutaminase: Monte Carlo analysis of small-angle scattering data. AB - Small-angle neutron and x-ray scattering experiments have been performed on type 2 tissular transglutaminase to characterize the conformational changes that bring about Ca(2+) activation and guanosine triphosphate (GTP) inhibition. The native and a proteolyzed form of the enzyme, in the presence and in the absence of the two effectors, were considered. To describe the shape of transglutaminase in the different conformations, a Monte Carlo method for calculating small-angle neutron scattering profiles was developed by taking into account the computer-designed structure of the native transglutaminase, the results of the Guinier analysis, and the essential role played by the solvent-exposed peptide loop for the conformational changes of the protein after activation. Although the range of the neutron scattering data is rather limited, by using the Monte Carlo analysis, and because the structure of the native protein is available, the distribution of the protein conformations after ligand interaction was obtained. Calcium activation promotes a rotation of the C-terminal with respect to the N-terminal domain around the solvent-exposed peptide loop that connects the two regions. The psi angle between the longest axes of the two pairs of domains is found to be above 50 degrees, larger than the psi value of 35 degrees calculated for the native transglutaminase. On the other hand, the addition of GTP makes possible conformations characterized by psi angles lower than 34 degrees. These results are in good agreement with the proposed enzyme activity regulation: in the presence of GTP, the catalytic site is shielded by the more compact protein structure, while the conformational changes induced by Ca(2+) make the active site accessible to the substrate. PMID- 10828002 TI - Direct observation of specific messenger RNA in a single living cell under a fluorescence microscope. AB - We observed the expression of human c-fos mRNA in a living transfected Cos7 cell under a fluorescence microscope by detecting hybrid formed with two fluorescently labeled oligodeoxynucleotides (oligoDNAs) and c-fos mRNA in the cytoplasm. Two fluorescent oligoDNAs were prepared, each labeled with a fluorescence molecule different from the other. When two oligoDNAs hybridized to an adjacent sequence on the target mRNA, the distance between the two fluorophores became very close and fluorescence resonance energy transfer (FRET) occurred, resulting in changes in fluorescence spectra. To find sequences of high accessibility of c-fos RNA to oligoDNAs, several sites that included loop structures on the simulated secondary structure were selected. Each site was divided into two halves, and the pair of fluorescent oligoDNAs complementary to the sequence was synthesized. Each site was examined for the efficiency of hybridization to c-fos RNA by measuring changes in fluorescence spectra when c-fos RNA was added to the pair of oligoDNAs in solution. A 40 mer specific site was found, and the pair of oligoDNAs for the site were microinjected into Cos7 cells that expressed c-fos mRNA. To block oligoDNAs from accumulating in the nucleus, oligoDNA was bound to a macromolecule (streptavidin) to prevent passage of nuclear pores. Hybridization of the pair of oligoDNAs to c-fos mRNA in the cytoplasm was detected in fluorescence images indicating FRET. PMID- 10828004 TI - Direct characterization of the physicochemical properties of fungal spores using functionalized AFM probes. AB - A new method is described for characterizing the physicochemical properties of native microbial cells by using atomic force microscopy (AFM) with chemically functionalized probes. Adhesion forces were measured, under deionized water, between probes and model substrata functionalized with alkanethiol self-assembled monolayers terminated with OH and CH(3) groups. These were found to be 6 +/- 2 nN (n = 1024), 0.9 +/- 0.4 nN, and approximately 0 nN, for CH(3)/CH(3), CH(3)/OH, and OH/OH surfaces, respectively, and were not significantly influenced by changes of ionic strength (0.1 M NaCl versus deionized water). This shows that functionalized probes are very sensitive to changes of surface hydrophobicity. Using OH- and CH(3)-terminated probes, patterns of rodlets, approximately 10 nm in diameter, were visualized, under physiological conditions, at the surface of spores of Phanerochaete chrysosporium. Multiple (1024) force-distance curves recorded over 500 x 500-nm areas at the spore surface, either in deionized water or in 0.1 M NaCl solutions, always showed no adhesion for both OH- and CH(3) terminated probes. Control experiments indicated that the lack of adhesion is not due to transfer of cellular material onto the probe, but to the hydrophilic nature of the spore surface. PMID- 10828003 TI - A metal-chelating microscopy tip as a new toolbox for single-molecule experiments by atomic force microscopy. AB - In recent years, the atomic force microscope (AFM) has contributed much to our understanding of the molecular forces involved in various high-affinity receptor ligand systems. However, a universal anchor system for such measurements is still required. This would open up new possibilities for the study of biological recognition processes and for the establishment of high-throughput screening applications. One such candidate is the N-nitrilo-triacetic acid (NTA)/His-tag system, which is widely used in molecular biology to isolate and purify histidine tagged fusion proteins. Here the histidine tag acts as a high-affinity recognition site for the NTA chelator. Accordingly, we have investigated the possibility of using this approach in single-molecule force measurements. Using a histidine-peptide as a model system, we have determined the binding force for various metal ions. At a loading rate of 0.5 microm/s, the determined forces varied from 22 +/- 4 to 58 +/- 5 pN. Most importantly, no interaction was detected for Ca(2+) and Mg(2+) up to concentrations of 10 mM. Furthermore, EDTA and a metal ion reloading step demonstrated the reversibility of the approach. Here the molecular interactions were turned off (EDTA) and on (metal reloading) in a switch-like fashion. Our results show that the NTA/His-tag system will expand the "molecular toolboxes" with which receptor-ligand systems can be investigated at the single-molecule level. PMID- 10828005 TI - Therapy-related acute myeloid leukemia and myelodysplasia after high-dose chemotherapy and autologous stem cell transplantation. AB - Therapy-related myelodysplasia (t-MDS) and acute myeloid leukemia (t-AML) after high-dose chemotherapy (HD-CT) and autologous stem cell transplantation (ASCT) for malignant diseases have become an important problem. The actuarial risk has varied, but has often been high if compared to the risk after conventional therapy. Prior chemotherapy with large cumulative doses of alkylating agents is the most important risk factor. In addition, patient age and previous radiotherapy, particularly the use of total body irradiation (TBI) in the preparative regimen for ASCT, have been identified as risk factors. In 3 studies, patients transplanted with CD34(+ )cells from peripheral blood after chemotherapy priming showed a higher risk of t-MDS or t-AML than patients transplanted with cells isolated from the bone marrow without priming. To what extent this higher risk relates to the prior therapy with a different contamination with preleukemic, hematopoietic precursors of the CD34(+) cells obtained by the 2 methods, or is a direct result of chemotherapy priming, or of an increasing awareness of these complications, remains to be determined. The latent period from ASCT to t-MDS and t-AML has often been short, 12 months or less in 27% of the patients. Bone marrow pathology of early cases of t-MDS after ASCT has often been neither diagnostic nor prognostic, but most patients presented chromosome aberrations, primarily deletions or loss of the long arms of chromosomes 5 and 7. The prognosis was in general poor, although 17% with indolent t-MDS survived more than 18 months from diagnosis, and most of these presented a normal karyotype or a single chromosome aberration. PMID- 10828006 TI - Translational pathophysiology: a novel molecular mechanism of human disease. AB - In higher eukaryotes, the expression of about 1 gene in 10 is strongly regulated at the level of messenger RNA (mRNA) translation into protein. Negative regulatory effects are often mediated by the 5'-untranslated region (5'-UTR) and rely on the fact that the 40S ribosomal subunit first binds to the cap structure at the 5'-end of mRNA and then scans for the first AUG codon. Self-complementary sequences can form stable stem-loop structures that interfere with the assembly of the preinitiation complex and/or ribosomal scanning. These stem loops can be further stabilized by the interaction with RNA-binding proteins, as in the case of ferritin. The presence of AUG codons located upstream of the physiological start site can inhibit translation by causing premature initiation and thereby preventing the ribosome from reaching the physiological start codon, as in the case of thrombopoietin (TPO). Recently, mutations that cause disease through increased or decreased efficiency of mRNA translation have been discovered, defining translational pathophysiology as a novel mechanism of human disease. Hereditary hyperferritinemia/cataract syndrome arises from various point mutations or deletions within a protein-binding sequence in the 5'-UTR of the L ferritin mRNA. Each unique mutation confers a characteristic degree of hyperferritinemia and severity of cataract in affected individuals. Hereditary thrombocythemia (sometimes called familial essential thrombocythemia or familial thrombocytosis) can be caused by mutations in upstream AUG codons in the 5'-UTR of the TPO mRNA that normally function as translational repressors. Their inactivation leads to excessive production of TPO and elevated platelet counts. Finally, predisposition to melanoma may originate from mutations that create translational repressors in the 5'-UTR of the cyclin-dependent kinase inhibitor 2A gene. PMID- 10828007 TI - The chemokine SDF-1 activates the integrins LFA-1, VLA-4, and VLA-5 on immature human CD34(+) cells: role in transendothelial/stromal migration and engraftment of NOD/SCID mice. AB - Hematopoietic stem cell homing and engraftment require several adhesion interactions, which are not fully understood. Engraftment of nonobese/severe combined immunodeficiency (NOD/SCID) mice by human stem cells is dependent on the major integrins very late activation antigen-4 (VLA-4); VLA-5; and to a lesser degree, lymphocyte function associated antigen-1 (LFA-1). Treatment of human CD34(+) cells with antibodies to either VLA-4 or VLA-5 prevented engraftment, and treatment with anti-LFA-1 antibodies significantly reduced the levels of engraftment. Activation of CD34(+) cells, which bear the chemokine receptor CXCR4, with stromal derived factor 1 (SDF-1) led to firm adhesion and transendothelial migration, which was dependent on LFA-1/ICAM-1 (intracellular adhesion molecule-1) and VLA-4/VCAM-1 (vascular adhesion molecule-1). Furthermore, SDF-1-induced polarization and extravasation of CD34(+)/CXCR4(+) cells through the extracellular matrix underlining the endothelium was dependent on both VLA-4 and VLA-5. Our results demonstrate that repopulating human stem cells functionally express LFA-1, VLA-4, and VLA-5. Furthermore, this study implies a novel approach to further advance clinical transplantation. PMID- 10828008 TI - Glycoprotein Ia gene C807T polymorphism and risk for major adverse cardiac events within the first 30 days after coronary artery stenting. AB - The glycoprotein complex Ia/IIa (GP Ia/IIa) is a major collagen receptor on platelets and other cell types. Recently, linked polymorphisms within the coding region of the GP Ia gene (C807T and G873A) were identified that are related to GP Ia/IIa surface expression. The T807/A873 allele is associated with high expression, whereas the C807/G873 allele is associated with low surface expression of GP Ia/IIa. Subsequently, the T807 allele was found to be associated with coronary and cerebral infarction in younger patients. Platelet adhesion to the vessel wall plays a pivotal role in thrombosis after coronary artery stent placement. The goal of this study was to test whether C807T polymorphism is associated with a higher incidence of thrombotic events following coronary stenting. Consecutive patients treated with coronary stent placement (n = 1797) were genotyped for C807T polymorphism with polymerase chain reaction and allele specific fluorogenic probes. The composite end point was defined as death, myocardial infarction, or urgent target vessel revascularization within 30 days of stent implantation. The genotype distribution of the study population was CC in 36.5%, CT in 46.7%, and TT in 16.8% of the patients. The incidence of the composite end point was 6.5% in T allele carriers and 5.3% in noncarriers (odds ratio for T allele carriage 1.23 [95% confidence interval, 0.81-1.86], P =.33). After adjusting for other baseline characteristics, the odds ratio for the composite end point was 1.15 (0.76-1.75). Therefore, C807T genotype has no significant influence on the major adverse events occurring after coronary artery stenting. PMID- 10828009 TI - Phase I/II trial of neutrophil transfusions from donors stimulated with G-CSF and dexamethasone for treatment of patients with infections in hematopoietic stem cell transplantation. AB - We examined the feasibility of a community blood bank granulocyte transfusion program utilizing community donors stimulated with a single-dose regimen of subcutaneous granulocyte colony-stimulating factor (G-CSF) plus oral dexamethasone. The recipients of these transfusions were neutropenic stem cell transplantation patients with severe bacterial or fungal infection. Nineteen patients received 165 transfusions (mean 8.6 transfusions/patient, range 1-25). Community donors provided 94% of the transfusions; relatives accounted for only 6% of the transfusions. Sixty percent of the community donors initially contacted agreed to participate, and 98% of these individuals indicated willingness to participate again. Transfusion of 81.9 +/- 2.3 x 10(9) neutrophils (mean +/- SD) resulted in a mean 1-hour posttransfusion neutrophil increment of 2. 6 +/- 2.6 x 10(3)/microL and restored the peripheral neutrophil count to the normal range in 17 of the 19 patients. The buccal neutrophil response, a measure of the capacity of neutrophils to migrate to tissue sites in vivo, was restored to normal in most patients following the transfusion. Chills, fever, and arterial oxygen desaturation of >/= 3% occurred in 7% of the transfusions, but these changes were not sufficient to limit therapy. Infection resolved in 8 of 11 patients with invasive bacterial infections or candidemia. These studies indicate that transfusion of neutrophils from donors stimulated with G-CSF plus dexamethasone can restore a severely neutropenic patient's blood neutrophil supply and neutrophil inflammation response. Further studies are needed to evaluate the clinical efficacy of this therapy. PMID- 10828010 TI - Improved outcome in childhood acute lymphoblastic leukemia despite reduced use of anthracyclines and cranial radiotherapy: results of trial ALL-BFM 90. German Austrian-Swiss ALL-BFM Study Group. AB - Trial ALL-BFM 90 was designed to improve outcome in patients with childhood acute lymphoblastic leukemia (ALL) by using a reduced treatment regimen. Patients were stratified into a standard-risk group (SRG), a medium-risk group (MRG), both defined by adequate early treatment response; and a high-risk group (HRG), defined by inadequate response to the cytoreductive prednisone prephase, induction failure, or Philadelphia-chromosome-positive ALL. Four treatment modifications were evaluated: dose intensification in induction by a more rapid drug sequence; administration of L-asparaginase during consolidation therapy in the MRG (randomized); enforced consolidation by rotational elements in the HRG; and reduction in the dose of anthracyclines and use of only 12-Gy preventive cranial radiotherapy in the MRG and HRG, with the aim of avoiding toxicity. Among all 2178 patients (/= 2 years), disease phase at transplant (first chronic phase vs other), disease stage at relapse (cytogenetic or chronic phase vs advanced phase), time from transplant to relapse (< 1 year vs >/= 1 year), and donor type (HLA-identical sibling vs volunteer unrelated donor). The effects of individual adverse risk factors were cumulative: The probability of survival at 10 years decreased stepwise from 42% (0 factors), 32% (1 factor), 14% (2 factors), 3% (3 factors), to 0% (4 or 5 factors). Novel strategies for high-risk patients are warranted. We conclude that these 5 factors should be taken into account when comparing results of salvage therapies in patients with Ph+ chronic myeloid leukemia relapsing after allogeneic stem cell transplant. PMID- 10828013 TI - Inhibition of granulocyte colony-stimulating factor-mediated myeloid maturation by low level expression of the differentiation-defective class IV granulocyte colony-stimulating factor receptor isoform. AB - In acute myeloid leukemia (AML), granulocyte colony-stimulating factor receptor (G-CSFR) proliferative and maturational signaling pathways are uncoupled. Seven human G-CSFR mRNA isoforms exist, named class I through class VII. The 183-amino acid cytosolic domain of the class I isoform provides all signaling activities. The class IV isoform is "differentiation defective" because the carboxy-terminal 87 amino acids are replaced with 34 amino acids of novel sequence. In more than 50% of AML samples, the class IV/class I G-CSFR mRNA ratio is aberrantly elevated compared to normal CD34(+) bone marrow cells. We hypothesized that the increased relative expression of class IV G-CSFR in AML uncouples proliferative and maturational G-CSFR signaling pathways. To test this, we transfected the G-CSF responsive murine cell line 32Dcl3 with class IV G-CSFR cDNA. After 10 days of G CSF stimulation, clones expressing class IV G-CSFR had greater percentages of myeloblasts and promyelocytes than controls (53% +/- 13% versus 3% +/- 2%). Differential counts over time demonstrated delayed G-CSF-driven maturation in 5 class IV-expressing clones, with 2 clones demonstrating a subpopulation that completely failed to differentiate. Heterologous class IV expression did not affect G-CSF-dependent proliferation. Class IV/murine G-CSFR mRNA ratios after 24 hours of G-CSF stimulation for 3 of the 5 clones (range, 0. 090 to 0.245; mean, 0.152 +/- 0.055) are within the range of class IV/class I mRNA ratios seen in patients with AML. This indicates that aberrantly increased relative class IV G CSFR expression seen in AML can uncouple G-CSFR proliferative and maturational signaling pathways. PMID- 10828014 TI - Identification and characterization of a new human ETS-family transcription factor, TEL2, that is expressed in hematopoietic tissues and can associate with TEL1/ETV6. AB - The ETS family of proteins is a large group of transcription factors implicated in many aspects of normal hematopoietic development, as well as oncogenesis. For example, the TEL1/ETV6 (TEL1) gene is required for normal yolk sac angiogenesis, adult bone marrow hematopoiesis, and is rearranged or deleted in numerous leukemias. This report describes the cloning and characterization of a novel ETS gene that is highly related to TEL1 and is therefore called TEL2. The TEL2 gene consists of 8 exons spanning approximately 21 kilobases (kb) in human chromosome 6p21. Unlike the ubiquitously expressed TEL1 gene, however, TEL2 appears to be expressed predominantly in hematopoietic tissues. Antibodies raised against the C terminus of the TEL2 protein were used to show that TEL2 localizes to the nucleus. All ETS proteins can bind DNA via the highly conserved ETS domain, which recognizes a purine-rich DNA sequence with a GGAA core motif. DNA binding assays show that TEL2 can bind the same consensus DNA binding sequence recognized by TEL1/ETV6. Additionally, the TEL2 protein is capable of associating with itself and with TEL1 in doubly transfected Hela cells, and this interaction is mediated through the pointed (PNT) domain of TEL1. The striking similarities of TEL2 to the oncogenic TEL1, its expression in hematopoietic tissues, and its ability to associate with TEL1 suggest that TEL2 may be an important hematopoietic regulatory protein. PMID- 10828015 TI - Vitamin A deficiency in mice causes a systemic expansion of myeloid cells. AB - To examine the role of retinoids in hematopoietic cell growth in vivo, we studied female SENCAR mice made vitamin A deficient by dietary restriction. Deficient mice exhibited a dramatic increase in myeloid cells in bone marrow, spleen, and peripheral blood. The abnormal expansion of myeloid cells was detected from an early stage of vitamin A deficiency and contrasted with essentially normal profiles of T and B lymphocytes. This abnormality was reversed on addition of retinoic acid to the vitamin A-deficient diet, indicating that the myeloid cell expansion is a direct result of retinoic acid deficiency. TUNEL analysis indicated that spontaneous apoptosis, a normal process in the life cycle of myeloid cells, was impaired in vitamin A-deficient mice, which may play a role in the increased myeloid cell population. Quantitative reverse transcriptase polymerase chain reaction analysis of purified granulocytes showed that expression of not only RAR, but RXRs, 2 nuclear receptors that mediate biologic activities of retinoids, was significantly reduced in cells of deficient mice. This work shows that retinoids critically control the homeostasis of myeloid cell population in vivo and suggests that deficiency in this signaling pathway may contribute to various myeloproliferative disorders. PMID- 10828016 TI - High-affinity binding to the GM-CSF receptor requires intact N-glycosylation sites in the extracellular domain of the beta subunit. AB - The human granulocyte-macrophage colony-stimulating factor (GM-CSF) receptor consists of 2 glycoprotein subunits, GMRalpha and GMRbeta. GMRalpha in isolation binds to GM-CSF with low affinity. GMRbeta does not bind GM-CSF by itself, but forms a high-affinity receptor in association with GMRalpha. Previously, it was found that N-glycosylation of GMRalpha is essential for ligand binding. The present study investigated the role of N-glycosylation of the beta subunit on GM CSF receptor function. GMRbeta has 3 potential N-glycosylation sites in the extracellular domain at Asn58, Asn191, and Asn346. Single mutants and triple mutants were constructed, converting asparagine in the target sites to aspartic acid or alanine. A single mutation at any of the 3 consensus N-glycosylation sites abolished high-affinity GM-CSF binding in transfected COS cells. Immunofluorescence and subcellular fractionation studies demonstrated that all of the GMRbeta mutants were faithfully expressed on the cell surface. Reduction of apparent molecular weight of the triple mutant proteins was consistent with loss of N-glycosylation. Intact N-glycosylation sites of GMRbeta in the extracellular domain are not required for cell surface targeting but are essential for high affinity GM-CSF binding. PMID- 10828017 TI - Intramedullary and extramedullary B lymphopoiesis in osteopetrotic mice. AB - Adult bone marrow is a major site for hematopoiesis, and reduction of the bone marrow cavity induces hematopoiesis in extramarrow tissues. To investigate the rudimentary intramarrow and the compensatory extramarrow hematopoiesis, particularly B lymphopoiesis, we used 3 osteopetrotic mouse strains [op/op, mi/mi, and Fos (-/-)], which are severely deficient in functional osteoclasts and therefore form inadequate bone marrow cavities. We found that bone marrow in these osteopetrotic mice supports myelopoiesis but not B lymphopoiesis, although cells that have the potential to differentiate into B lineage cells are present in the bone marrow. Although B lymphopoiesis normally occurs both in the spleen and liver of newborn mice, compensatory B lymphopoiesis in adult op/op and mi/mi mice is observed only in the liver, while myelopoiesis is enhanced in both organs. Interestingly, mice lacking the Fos proto-oncogene exhibit B lymphopoiesis in the spleen as well as liver. The amounts of expression of steel factor, Flt3/Flk-2 ligand, and interleukin-7 in the bone marrow, spleen, or liver were not significantly affected in these osteopetrotic mutants. These findings suggest that the volume of the bone marrow cavity regulates B lymphopoiesis without affecting the production of certain hematopoietic growth factors. The splenic microenvironments that support both myelopoiesis and B lymphopoiesis in the neonatal stage are lost in adults and are not reactivated even in the osteopetrotic adults unless the Fos gene is disrupted. PMID- 10828018 TI - Smad7 selectively interferes with different pathways of activin signaling and inhibits erythroid leukemia cell differentiation. AB - Smad family proteins are essential for transforming growth factor beta (TGF-beta) signal mediation downstream of a heteromeric complex of the type I and type II receptor serine/threonine kinases. A distant family member, Smad7, is expressed in most mammalian tissues and cells and prevents TGF-beta signaling. In this study, we examined the physiologic role of Smad7 in mediating the effects of activin, a member of the TGF-beta superfamily of peptides that functions in a number of processes, including blood-cell development. We report here that Smad7 expression is specifically absent in particular hematopoietic cells that respond to activin by differentiating into the erythroid lineage and that ectopic production of Smad7 causes mouse erythroid leukemia (F5-5) cells to become resistant to activin induction of erythroid differentiation. When coexpressed with type I activin receptor ActR-I or ActR-IB in concert with type II receptor ActR-II, Smad7 efficiently reduced an early transcriptional response mediated by ActR-I but had only a minimal effect on the response mediated by ActR-IB. In the presence of Smad7, overexpression of an activated form of ActR-IB, but not of an activated form of ActR-I, induced F5-5 cells to differentiate. These results suggest that Smad7 selectively interferes with the ActR-I pathway in activin signal transduction. The findings also indicate the existence of a novel activity of Smad7 that inhibits erythroid differentiation by blocking intracellular signaling of activin. PMID- 10828019 TI - Conserved amino acids in metal-binding motifs of PDE3A are involved in substrate and inhibitor binding. AB - The activity of phosphodiesterase (PDE)3A requires divalent cations. Putative metal-binding sites are expected at 2 highly conserved metal-binding motifs, HXXXH(X)(25)E. A functional truncated recombinant PDE3A containing the catalytic domain (PDE3Atriangle up1) and mutant proteins were expressed in a baculovirus/Sf9 cell system. All the mutant proteins had decreased catalytic efficiency (k(cat)/K(m)). Mutants H752A, H756A, and E825A had k(cat) of less than 0.0008 s(-1) to 0.0475 s(-1) compared to PDE3Atriangle up1, with 1.86 second(-1), with unchanged K(m). Although E866A had a k(cat) of 0.235 s(-1), the K(m) for cyclic adenosine monophosphate (cAMP) was increased 11-fold and the K(i) for cyclic guanosine monophosphate (cGMP) was 27-fold higher than PDE3Atriangle up1. The K(i) of H836A for cGMP was 177-fold higher than that of PDE3Atriangle up1. The K(m) for E971A was 5-fold higher than PDE3Atriangle up1. These results suggest that the cAMP and cGMP binding sites are overlapping, but not identical, involving both common and different amino acids. Mutants E825A, H836A, and E866A showed low activity in a metal ion-free assay; however, their enzymatic activities were increased 4- to 10-fold in buffers containing Mn(2+), Mg(2+), or Co(2+). This observation indicates that conserved amino acids in the second metal binding motif might not be involved in binding divalent cations but may serve other functions such as substrate or inhibitor binding in PDE3A. PMID- 10828020 TI - Up-regulation of vascular endothelial growth factor receptor Flt-1 after endothelial denudation: role of transcription factor Egr-1. AB - Vascular endothelial growth factor (VEGF) is highly expressed in vascular remodeling processes and accelerates reendothelialization after mechanical denudation. Two VEGF tyrosine kinase receptors have been reported-fms-like tyrosine kinase-1 (Flt-1) and kinase domain region (KDR). Little is known about the regulation of the expression of these receptors after vascular injury. Herein, we have analyzed the expression of Flt-1 after mechanical denudation of primary cultures of endothelial cells, which has been considered a useful in vitro model to study endothelium responses to vascular injury. After denudation, the Flt-1 protein and mRNA levels are clearly up-regulated, and transient transfection experiments showed a strong induction of the flt-1 promoter dependent transcription. Analysis of the flt-1 promoter sequence revealed the presence of a putative binding site for the early growth response factor-1 (Egr 1) at positions -24 to -16. Electrophoretic mobility shift and supershift assays showed that Egr-1 was able to bind to this DNA sequence, and cotransfection of the flt-1 promoter reporter plasmid with an Egr-1 expression vector resulted in enhancement of its transcriptional activity. Furthermore, the mutation of the Egr 1 binding site markedly reduced the denudation-induced flt-1 promoter activity. These data demonstrate that Flt-1 is up-regulated after endothelial denudation and that Egr-1 plays a relevant role in this process. PMID- 10828021 TI - Binding and transfer of verocytotoxin by polymorphonuclear leukocytes in hemolytic uremic syndrome. AB - The hemolytic uremic syndrome (HUS) is the most common cause of acute renal failure in children. The role of a verocytotoxin (VT)-producing Escherichia coli has been strongly implicated in the epidemic form of HUS. Although direct toxicity of VT on glomerular endothelial cells has been demonstrated, it remained still unclear how the VT is transported from the intestine to the target organs. In this study we demonstrate that VT, when incubated in whole blood, binds rapidly and completely to human polymorphonuclear leukocytes (PMNs) and not to other components of blood. Binding studies with (125)I-VT-1 showed a single class of binding sites on freshly isolated, nonstimulated human PMNs. The K(d) of VT binding to PMNs was 10(-8) mol/L, 100-fold less than that of the VT-receptor globotriaosylceramide. On incubation of VT-preloaded PMNs with human glomerular microvascular endothelial cells (GMVECs), transfer of VT-1 to the endothelial cells occurred. Incubation of nonstimulated GMVECs with VT-preloaded PMNs, but not with PMNs or VT-1 alone, caused inhibition of protein synthesis and cell death. Our data are in concert with a role of PMNs in the transfer of VT from the intestine to the kidney endothelium. This transfer occurs by selective binding to a specific receptor on PMNs and subsequent passing of the ligand VT to the VT receptor on GMVECs, which causes cell damage. This new mechanism further underpins the important role of PMNs in HUS. PMID- 10828022 TI - Endostatin-induced tyrosine kinase signaling through the Shb adaptor protein regulates endothelial cell apoptosis. AB - Endostatin, which corresponds to the C-terminal fragment of collagen XVIII, is a potent inhibitor of angiogenesis. Fibroblast growth factor-2 (FGF-2)-induced angiogenesis in the chicken chorioallantoic membrane was inhibited by endostatin, but not by an endostatin mutant R158/270A, lacking heparin-binding ability. Endostatin was internalized by endothelial cells, but not by mouse fibroblasts. Treatment of murine brain endothelial (IBE) cells with endostatin reduced the proportion of cells in S phase, whereas growth-arrested IBE cells in collagen gels treated with endostatin displayed enhanced tubular morphogenesis. IBE cells overexpressing Shb, an adaptor protein implicated in angiostatin-induced apoptosis, displayed elevated apoptosis and decreased tubular morphogenesis in collagen gels in response to endostatin when added together with FGF-2. Induction of apoptosis was dependent on the heparin-binding ability of endostatin and the expression of Shb with a functional Src homology 2 (SH2)-domain. Endostatin treatment for 10 minutes or 24 hours induced tyrosine phosphorylation of Shb and formation of multiprotein complexes. An Shb SH2 domain fusion protein precipitated a 125-kd phosphotyrosyl protein in endostatin-treated cells. The 125 kd component either contained intrinsic tyrosine kinase activity or occurred in complex with a tyrosine kinase. In conclusion, our data show that endostatin induces tyrosine kinase activity and enhanced apoptosis in FGF-treated endothelial cells. PMID- 10828023 TI - Distinct localization and function of (1,4,5)IP(3) receptor subtypes and the (1,3,4,5)IP(4) receptor GAP1(IP4BP) in highly purified human platelet membranes. AB - Platelet activation is associated with an increase of cytosolic Ca(++) levels. The (1,4,5)IP(3) receptors [(1,4,5)IP(3)R] are known to mediate Ca(++) release from intracellular stores of many cell types. Currently there are at least 3 distinct subtypes of (1,4, 5)IP(3)R-type I, type II, and type III-with suggestions of distinct roles in Ca(++) elevation. Specific receptors for (1,3,4,5)IP(4) belonging to the GAP1 family have also been described though their involvement with Ca(++) regulation is controversial. In this study we report that platelets contain all 3 subtypes of (1,4,5)IP(3)R but in different amounts. Type I and type II receptors are predominant. In studies using highly purified platelet plasma (PM) and intracellular membranes (IM) we report a distinct localization of these receptors. The PM fractions were found to contain the type III (1,4,5)IP(3)R and GAP1(IP4BP) in contrast to IM, which contained type I (1,4,5)IP(3)R. The type II receptor exhibited a dual distribution. In studies examining the labeling of surface proteins with biotin in intact platelets only the type III (1,4,5)IP(3)R was significantly labeled. Immunogold studies of ultracryosections of human platelets showed significantly more labeling of the PM with the type III receptor antibodies than with type I receptor antibodies. Ca(++) flux studies were carried out with the PM to demonstrate in vitro function of inositol phosphate receptors. Ca(++) release activities were present with both (1,4,5)IP(3) and (1, 3,4,5)IP(4) (EC(50) = 1.3 and 0.8 micromol/L, respectively). Discrimination of the Ca(++)-releasing activities was demonstrated with cyclic adenosine monophosphate (cAMP)-dependent protein kinase (cAMP-PK) specifically inhibiting (1,4,5)IP(3) but not (1,3,4, 5)IP(4)-induced Ca(++) flux. In experiments with both PM and intact platelets, the (1,4,5)IP(3)Rs but not GAP1(IP4BP) were found to be substrates of cAMP-PK and cGMP-PK. Thus the Ca(++) flux property of (1,3,4,5)IP(4) is insensitive to cAMP-PK. These studies suggest distinct roles for the (1,4,5)IP(3)R subtypes in Ca(++) movements, with the type III receptor and GAP1(IP4BP) associated with cation entry in human platelets and the type I receptor involved with Ca(++) release from intracellular stores. PMID- 10828024 TI - Modulation of factor VII levels by intron 7 polymorphisms: population and in vitro studies. AB - Previous studies have established that factor VII gene (F7) polymorphisms (5'F7 and R353Q) contribute about one-third of factor VII (FVII) level variation in plasma. However, F7 genotyping in patients with cardiovascular disease has produced conflicting results. Population and expression studies were used to investigate the role of intron 7 (IVS7 ) polymorphisms, including repeat and sequence variations, in controlling activated FVII (FVIIa) and antigen (FVIIag) levels. Genotype-phenotype studies performed in 438 Italian subjects suggested a positive relation between the IVS7 repeat number and FVII levels. The lowest values were associated with the IVS7 + 7G allele. The screening of 52 patients with mild FVII deficiency showed an 8-fold increase in frequency (8%) of this allele, and among heterozygotes for identical mutations, lower FVII levels were observed in the IVS7 + 7G carriers. This frequent genetic component participates in the phenotypic heterogeneity of FVII deficiency. The evaluation of the individual contribution of polymorphisms was assisted by the expression of each IVS7 variant, as a minigene, in eukaryotic cells. The novel quantitative analysis revealed that higher numbers of repeats were associated with higher mRNA expression levels and that the IVS7 + 7G allele, previously defined as a functionally silent polymorphism, was responsible for the lowest relative mRNA expression. Taken together, these findings indicate that the IVS7 polymorphisms contribute to the plasmatic variance of FVII levels via differential efficiency of mRNA splicing. These studies provide further elements to understand the control of FVII levels, which could be of importance to ensure the hemostatic balance under pathologic conditions. PMID- 10828025 TI - Thrombopoietin potentiates collagen receptor signaling in platelets through a phosphatidylinositol 3-kinase-dependent pathway. AB - Collagen activates platelets through a tyrosine kinase-dependent pathway, involving phospholipase Cgamma2. Functional responses such as aggregation and secretion induced by collagen are potentiated by preincubation with thrombopoietin (TPO). In this study, we show that collagen and thrombopoietin activate the phosphatidylinositol 3-kinase (PI 3-kinase) pathway and that this contributes to their respective actions. The structurally distinct inhibitors of PI 3-kinase, wortmannin, and LY294002, completely inhibit formation of phosphatidylinositol 3,4,5-trisphosphate by collagen. This leads to a substantial reduction in the formation of inositol phosphates and phosphatidic acid, 2 indices of PLC activity, and the consequent inhibition of intracellular Ca(++) [Ca(++)](i), aggregation and secretion. Potentiation of the collagen response by TPO is prevented in the presence of wortmannin and LY294002. However, when the 2 PI 3-kinase inhibitors are given after the addition of TPO but before the collagen, recovery of potentiation is observed. This suggests that potentiation is mediated through activation of PI 3-kinase. TPO stimulates aggregation of platelets from a low percentage of donors and this is also blocked by wortmannin. These results suggest that the PI 3-kinase pathway plays an important role in signaling by collagen and in the priming action of TPO. PMID- 10828026 TI - Antibodies to the FVIII light chain that neutralize FVIII procoagulant activity are present in plasma of nonresponder patients with severe hemophilia A and in normal polyclonal human IgG. AB - We have analyzed the properties of anti-factor VIII (FVIII) immunoglobulin (Ig) G recovered by affinity chromatography on FVIII-Sepharose from the IgG fraction of the plasma of healthy individuals and nonresponder patients with hemophilia A. Affinity-purified anti-FVIII antibodies were found to neutralize FVIII activity and to bind to FVIII with an affinity similar to that of anti-FVIII IgG that had been affinity-purified from the plasma of inhibitor-positive hemophilia patients and of patients with anti-FVIII autoimmune disease. The antibodies also exhibited patterns of reactivity with thrombin-digested FVIII similar to those of FVIII inhibitors and preferentially recognized epitopes located in the light chain of FVIII. These observations suggest that FVIII inhibitors occurring in hemophilia A and in patients with anti-FVIII autoimmune disease originate from the expansion of preexisting natural anti-FVIII clones that exhibit FVIII-neutralizing properties. PMID- 10828027 TI - Ferriporphyrins and endothelium: a 2-edged sword-promotion of oxidation and induction of cytoprotectants. AB - Heme arginate infusions blunt the symptoms of patients with acute intermittent porphyria without evidence of the vascular or thrombotic side effects reported for hematin. To provide a rationale for heme arginate's safety, the present study examined the effects of various ferriporphyrins to sensitize human endothelial cells to free radical injury and to induce heme oxygenase and ferritin expression. Heme arginate, unlike hematin, did not amplify oxidant-induced cytotoxicity mediated by hydrogen peroxide (5.3 +/- 2.4 versus 62.3 +/- 5.3% (51)Cr release, P <.0001) or by activated neutrophils (14.4 +/- 2.9 versus 41.1 +/- 6.0%, P <.0001). Nevertheless, heme arginate efficiently entered endothelial cells similarly to hematin, since both markedly induced heme oxygenase mRNA (more than 20-fold increase) and enzyme activity. Even with efficient permeation, endothelial cell ferritin content was only minimally increased by heme arginate compared with a 10-fold induction by hematin; presumably less free iron was derived from heme arginate despite up-regulation of heme oxygenase. Hematin is potentially vasculopathic by its marked catalysis of oxidation of low-density lipoprotein (LDL) to endothelial-toxic moieties. Heme arginate was significantly less catalytic. Heme arginate-conditioned LDL was less than half as cytotoxic to endothelial cells as hematin-conditioned LDL (P <.004). It is concluded that heme arginate may be less vasculotoxic than hematin since it is an effective heme oxygenase gene regulator but a less efficient free-radical catalyst. PMID- 10828028 TI - Nitric oxide induces apoptosis in megakaryocytic cell lines. AB - Cytokines that stimulate inducible nitric oxide (NO) synthase can suppress the growth and differentiation of normal human bone marrow cells, including megakaryocytes. Since NO promotes apoptosis in other cell systems, we chose to study the determinants of apoptosis in megakaryocytic cells. We show that both exogenous and endogenous sources of NO can induce apoptosis in megakaryocytoid cell lines. The megakaryocyte growth factor thrombopoietin suppresses NO-induced apoptosis, whereas treatment with peroxynitrite, a cytotoxic product formed when NO reacts with superoxide, promotes apoptosis. Superoxide inhibitors suppress NO induced apoptosis, and pretreatment with megakaryocyte growth and maturation factors attenuates NO-induced apoptosis. These data show that NO modulates megakaryocyte apoptosis and suggest that this process may occur in the cytokine rich marrow milieu to regulate megakaryocyte turnover. PMID- 10828029 TI - Hydroxychloroquine inhibits calcium signals in T cells: a new mechanism to explain its immunomodulatory properties. AB - Hydroxychloroquine (HCQ), a lysosomotropic amine, is an immunosuppressive agent presently being evaluated in bone marrow transplant patients to treat graft versus-host disease. While its immunosuppressive properties have been attributed primarily to its ability to interfere with antigen processing, recent reports demonstrate HCQ also blocks T-cell activation in vitro. To more precisely define the T-cell inhibitory effects of HCQ, the authors evaluated T-cell antigen receptor (TCR) signaling events in a T-cell line pretreated with HCQ. In a concentration-dependent manner, HCQ inhibited anti-TCR-induced up-regulation of CD69 expression, a distal TCR signaling event. Proximal TCR signals, including inductive protein tyrosine phosphorylation, tyrosine phosphorylation of phospholipase C gamma1, and total inositol phosphate production, were unaffected by HCQ. Strikingly, anti-TCR-crosslinking-induced calcium mobilization was significantly inhibited by HCQ, particularly at the highest concentrations tested (100 micromol/L) in both T-cell lines and primary T cells. HCQ, in a dose dependent fashion, also reduced a B-cell antigen receptor calcium signal, indicating this effect may be a general property of HCQ. Inhibition of the calcium signal correlated directly with a reduction in the size of thapsigargin sensitive intracellular calcium stores in HCQ-treated cells. Together, these findings suggest that disruption of TCR-crosslinking-dependent calcium signaling provides an additional mechanism to explain the immunomodulatory properties of HCQ. PMID- 10828030 TI - Heavy and light chain primary structures control IgG3 nephritogenicity in an experimental model for cryocrystalglobulinemia. AB - Crystal formation by monoclonal immunoglobulins is a well-known but rare complication of B-cell neoplasia. We have designed an in vivo model of cryocrystalglobulinemia by grafting to mice hybridoma clones producing a pathogenic monoclonal immunogloblulin (Ig) G3kappa. One clone, 8A4, secreted a singular IgG3 that formed crystals both in the proliferating plasma cells and as mesangial and subendothelial deposits in the kidney glomeruli. Morphologic analysis of kidneys revealed neutrophil infiltration and endocapillary hyperplasia, while the morphology of deposits was reminiscent of those in cryocrystalglobulinemia patients. A variant clone that only differed from 8A4 by a 3-amino acid deletion in the V(kappa) CDR1 increased its secretion level by 7 fold and produced an abundant bona fide serum monoclonal cryoglobulin in mice, without crystal formation within tumoral cells; it yielded no subendothelial deposits but only amorphous precipitates in capillary lumens of kidney glomeruli, reminiscent of those seen in the human hyperviscosity syndrome, without other glomerular lesions. A limited variation in the V(kappa) domain thus proved able to increase secretion, to abrogate crystallization, and to modify patterns of glomerular lesions and deposits. Both the crystallizing and noncrystallizing IgG3kappa sequences were related to previously reported murine cryoglobulins, all including a gamma3 chain and canonical VH sequences. Two additional variants of 8A4 with identical VH and VL domains but having switched to IgG1 also lost crystal formation, further showing this feature of 8A4 to result from a unique 3 dimensional conformation of the complete immunoglobulin, relying on V and C domain primary structures of both chains. PMID- 10828031 TI - Acquisition of intact allogeneic human leukocyte antigen molecules by human dendritic cells. AB - In an attempt to transduce monocyte-derived dendritic cells (DCs) by a retroviral vector coding for a cell surface marker, we were confronted by the observation of high transfer of the surface molecule in the absence of vector proviral DNA in the treated cells. Indeed, DCs acquired the surface marker by a mechanism independent of the vector machinery, requiring cell-to-cell contact and involving transfer of lipids and a variety of intact membrane proteins. Most important, this property of DCs also includes acquisition of foreign human leukocyte antigen (HLA) molecules. Consequently, DCs become immunological hybrids as they display their own and foreign HLA molecules. The newly acquired HLA is fully functional because it allows recognition by allo-specific T lymphocytes and the binding and presentation of antigen peptides. PMID- 10828032 TI - Expression of c-FLIP(L) and resistance to CD95-mediated apoptosis of monocyte derived dendritic cells: inhibition by bisindolylmaleimide. AB - To gain insight into the mechanisms controlling apoptosis of dendritic cells (DC), human monocyte-derived DC were analyzed for their expression of CD95 (Fas/Apo-1) and their response to CD95 ligation. Although DC expressed the CD95 molecule on their membrane, they did not undergo apoptosis on CD95 ligation unless sensitized by cycloheximide. In parallel, DC synthesized c-FLIP(L), an inhibitor of the CD95-mediated death-signaling cascade. We also demonstrated that bisindolylmaleimide down-regulates c-FLIP(L) expression in DC and, in parallel, allows CD95-mediated apoptosis in these cells. In contrast, Bcl-2, Bcl-x(L), and Bax levels were not affected by bisindolylmaleimide. We conclude that DC resist CD95- mediated apoptosis in association with c-FLIP(L) expression and that the immunosuppressive potential of bisindolylmaleimide previously observed at the T cell level also involves facilitation of CD95-mediated DC apoptosis. PMID- 10828033 TI - Failure of gelsolin overexpression to regulate lymphocyte apoptosis. AB - The actin regulatory protein gelsolin cleaves actin filaments in a calcium- and polyphosphoinositide-dependent manner. Gelsolin has recently been described as a novel substrate of the cysteinyl protease caspase-3, an effector protease activated during apoptosis. Cleavage by caspase-3 generates an amino-terminal fragment of gelsolin that can sever actin filaments independently of calcium regulation. The disruption of the actin cytoskeleton by cleaved gelsolin is hypothesized to mediate many of the downstream morphological changes associated with apoptosis. In contrast, overexpression of full-length gelsolin has also been reported to inhibit apoptotic cell death upstream of the activation of caspase-3, suggesting that gelsolin may also act prior to commitment to cell death. The authors previously observed that actin stabilization by the cell permeant agent jasplakinolide enhanced cell death upon interleukin (IL)-2 or IL-3 withdrawal from growth-factor-dependent lymphocyte cell lines, and hypothesized that actin polymerization could alter the activity of gelsolin, thus enhancing apoptosis. Here the authors show that constitutive overexpression of gelsolin did not, however, inhibit or dramatically enhance apoptotic cell death upon growth-factor withdrawal, nor did it modify sensitivity to jasplakinolide. In contrast to previous reports, overexpression of gelsolin in Jurkat T cells did not prevent or delay apoptosis induced by Fas ligation or ceramide treatment. Overexpressed gelsolin protein was cleaved during apoptosis, as seen previously in this and other cell types. In these model systems, therefore, the level of gelsolin expression was not a rate-limiting determinant in commitment to or time to the morphological changes of apoptosis. PMID- 10828034 TI - Mice lacking flt3 ligand have deficient hematopoiesis affecting hematopoietic progenitor cells, dendritic cells, and natural killer cells. AB - The ligand for the receptor tyrosine kinase fms-like tyrosine kinase 3 (flt3), also referred to as fetal liver kinase-2 (flk-2), has an important role in hematopoiesis. The flt3 ligand (flt3L) is a growth factor for hematopoietic progenitors and induces hematopoietic progenitor and stem cell mobilization in vivo. In addition, when mice are treated with flt3L immature B cells, natural killer (NK) cells and dendritic cells (DC) are expanded in vivo. To further elucidate the role of flt3L in hematopoiesis, mice lacking flt3L (flt3L-/-) were generated by targeted gene disruption. Leukocyte cellularity was reduced in the bone marrow, peripheral blood, lymph nodes (LN), and spleen. Thymic cellularity, blood hematocrit, and platelet numbers were not affected. Significantly reduced numbers of myeloid and B-lymphoid progenitors were noted in the BM of flt3L-/- mice. In addition a marked deficiency of NK cells in the spleen was noted. DC numbers were also reduced in the spleen, LN, and thymus. Both myeloid-related (CD11c(++) CD8alpha(-)) and lymphoid-related (CD11c(++) CD8alpha(+)) DC numbers were affected. We conclude that flt3L has an important role in the expansion of early hematopoietic progenitors and in the generation of mature peripheral leukocytes. PMID- 10828035 TI - Mechanism of resistance to the ABL tyrosine kinase inhibitor STI571 in BCR/ABL transformed hematopoietic cell lines. AB - The tyrosine kinase activity of the Bcr/Abl oncogene is required for transformation of hematopoietic cells. The tyrosine kinase inhibitor STI571 (formerly called CGP57148B, Novartis Pharmaceuticals) inhibits BCR/ABL, TEL/ABL, and v-ABL kinase activity and inhibits growth and viability of cells transformed by any of these ABL oncogenes. Here we report the generation of 2 BCR/ABL positive cell lines that have developed partial resistance to STI571. BCR/ABL transformed Ba/F3 hematopoietic cells and Philadelphia-positive human K562 cells were cultured in gradually increasing concentrations of STI571 over a period of several months to generate resistant lines. Resistant Ba/F3.p210 cells were found to have an increase in Bcr/Abl messenger RNA, amplification of the Bcr/Abl transgene, and a greater than tenfold increase in the level of BCR/ABL protein. In contrast to Ba/F3.p210 cells, drug-resistant K562 cells did not undergo detectable amplification of the BCR/ABL gene, although they displayed a 2-fold to 3-fold increase in p210BCR/ABL protein. The addition of STI571 to both resistant Ba/F3. p210 and K562 cells resulted in a rapid reduction of tyrosine phosphorylation of cellular proteins, similar to that observed for nonresistant cells. However, the inhibition of kinase activity was transient and partial and was not accompanied by apoptosis. The results suggest that resistance to STI571 may be multifactorial. Increased expression of the target protein BCR/ABL was observed in both lines, and resulted from oncogene amplification in one line. However, altered drug metabolism, transport, or other related mechanisms may also contribute to drug resistance. PMID- 10828036 TI - Interleukin-13 fusion cytotoxin as a potent targeted agent for AIDS-Kaposi's sarcoma xenograft. AB - Clinically advanced and rapidly progressive AIDS-associated Kaposi sarcoma (AIDS KS) tumors require an aggressive tumor-directed therapy. We have observed that AIDS-KS cells express high levels of receptors for immune regulatory cytokine, interleukin-13 (IL-13). Two tumorigenic AIDS-KS cell lines, KS Y-1 and KS-imm, expressed 4560 and 9480 IL-13 binding sites per cell with an affinity (kd) of approximately 0.9 and 3.7 nmol/L, respectively. IL-13 cytotoxin IL13-PE38QQR, consisting of human IL-13 and a derivative of Pseudomonas exotoxin, is specifically cytotoxic to KS tumor cells. Systemic and loco regional administration of IL13-PE38QQR in immunodeficient mice with established human KS tumors produced remarkable antitumor activity. Three intratumoral (IT) injections of IL-13 toxin (250 microg/kg per dose) on alternate days (qod) or 5 daily (qd) IT injections with lower doses (50 or 100 microg/kg per dose) resulted in a complete regression of established subcutaneous tumors in most animals. Daily IT treatment with 250 microg/kg of IL-13 toxin in another KS-derived cell line also produced complete responses. Twice daily intraperitoneal injections of IL13 PE38QQR (25 or 50 microg/kg per dose) for 10 days (total injections = 20) also completely eradicated KS Y-1 tumors. Intravenous administration of IL13-PE38QQR also suppressed tumor growth; however, complete responses were not observed. All animals tolerated the therapeutic doses of IL-13 toxin without any visible signs of toxicity. The efficacy of receptor-directed IL13-PE38QQR therapy in mice warrants further exploration of this drug for AIDS-KS treatment. PMID- 10828037 TI - Deletion of the multidrug resistance protein MRP1 gene in acute myeloid leukemia: the impact on MRP activity. AB - Deletion of the multidrug resistance gene MRP1 has been demonstrated in acute myeloid leukemia (AML) patients with inversion of chromosome 16 (inv[16]). These AML patients are known to have a relatively favorable prognosis, which suggests that MRP1 might play an important role in determining clinical outcome. This study analyzed MRP1 deletion by fluorescent in situ hybridization (FISH), with a focus on inv(16) AML patients. Functional activity of multidrug resistance protein (MRP) was studied in a flow cytometric assay with the use of the MRP substrate carboxyfluorescein (CF) and the inhibitor MK-571. MRP1, MRP2, and MRP6 messenger RNA (mRNA) expression was determined with reverse transcriptase polymerase chain reaction (RT-PCR). The results were compared with normal bone marrow cells. MRP1 deletion was detected in 7 AML patients; 2 cases showed no MRP1 FISH signals, and 5 cases had 1 MRP1 signal, whereas in 4 AML patients with inv(16) no MRP1 deletions were observed. A variability in MRP activity, expressed as CF efflux-blocking by MK-571, was observed (efflux-blocking factors varied between 1.2 and 3.6); this correlated with the number of MRP1 genes (r = 0.91, P <. 01). MRP activity in the AML cases was not different from normal hematopoietic cells. MRP1 mRNA was detected in patients with 1 or 2 MRP1 FISH signals, but not in patients with no MRP1 signals. MRP2 and MRP6 mRNA were expressed predominantly in AML samples with 1 MRP1 signal, whereas in normal bone marrow cells no MRP2 and MRP6 mRNA was observed. In conclusion, this study shows that MRP activity varies among inv(16) AML cases and does not differ from that in normal hematopoietic cells; this might be in part due to the up-regulation of other MRP genes. PMID- 10828038 TI - Follicular lymphomas' BCL-2/IgH junctions contain templated nucleotide insertions: novel insights into the mechanism of t(14;18) translocation. AB - The human t(14;18) chromosomal translocation is assumed to result from illegitimate rearrangement between BCL-2 and D(H)/J(H) gene segments during V(D)J recombination in early B cells. De novo nucleotides are found inserted in most breakpoints and have been thus far interpreted as nontemplated N region additions. In this report, we have analyzed both direct (BCL-2/J(H)) and reciprocal (D(H)/BCL-2) breakpoints derived from 40 patients with follicular lymphoma with t(14;18). Surprisingly, we found that more than 30% of the breakpoint junctions contain a novel type of templated nucleotide insertions, consisting of short copies of the surrounding BCL-2, D(H), and J(H) sequences. The features of these templated nucleotides, including multiplicity of copies for 1 template and the occurrence of mismatches in the copies, suggest the presence of a short-patch DNA synthesis, templated and error-prone. In addition, our analysis clearly shows that t(14;18) occurs during a very restricted window of B cell differentiation and involves 2 distinct mechanisms: V(D)J recombination, mediating the breaks on chromosome 14 during an attempted secondary D(H) to J(H) rearrangement, and an additional unidentified mechanism creating the initial breaks on chromosome 18. Altogether, these data suggest that the t(14;18) translocation is a more complex process than previously thought, involving the interaction and/or subversion of V(D)J recombination with multiple enzymatic machineries. PMID- 10828039 TI - Expression of gene MAGE-A4 in Reed-Sternberg cells. AB - Genes of the MAGE-A family are expressed in several types of solid tumors but are silent in normal tissues with the exception of male germline cells, which do not carry HLA molecules.Therefore, peptides encoded by MAGE-A genes are strictly tumor-specific antigens that can be recognized by CTL and constitute promising targets for immunotherapy. The expression of 6 genes of the MAGE-A family was tested with reverse transcriptase-polymerase chain reaction in lymphoma samples. Among 38 samples of non-Hodgkin lymphoma, 1 anaplastic large cell lymphoma expressed genes MAGE-A1, -A2, -A3, -A4, and -A12, and 1 lymphoepithelioid T-cell lymphoma expressed gene MAGE-A4. Five of 18 samples (28%) from patients with Hodgkin disease expressed gene MAGE-A4. In tissue sections, staining by a monoclonal antibody that recognizes the MAGE-A4 protein was observed in 11 of 53 samples (21%) from patients with Hodgkin disease. In the positive samples, the Reed-Sternberg cells were strongly stained whereas the surrounding cells were not. These results indicate that Hodgkin disease may be a target for specific immunotherapy involving MAGE-A4 antigens. PMID- 10828040 TI - Somatic mutation of bcl-6 genes can occur in the absence of V(H) mutations in chronic lymphocytic leukemia. AB - Somatic mutation in immunoglobulin variable (V) region genes occurs largely in the germinal center and, after neoplastic transformation, imprints V genes of B cell tumors with the mutational history of the cell of origin. Recently, it has been found that chronic lymphocytic leukemia (CLL) consists of 2 subsets, each with a different clinical course, one with unmutated V(H) genes consistent with a naive B cell, and the other with mutated V(H) genes consistent with transit through the germinal center. However, somatic mutation also occurs at another distinct locus, the 5' noncoding region of the bcl-6 gene, in both B-cell tumors and in normal germinal center B cells. To probe the suggestive link between the occurrence of mutations in V(H) and bcl-6 genes, we analyzed the nature of somatic mutation at these distinct loci in the 2 CLL subsets. Unexpectedly, we found no such link in the CLLs defined by unmutated V(H) genes, with 4 of 10 cases clearly showing mutations in bcl-6. In those CLLs defined by somatically mutated V(H) genes, 4 of 9 cases predictively showed bcl-6 mutations. The frequency of bcl-6 mutations was comparable in both subsets, with mutations being biallelic, and in 3 of 8 cases indicative of clonal origins. Surprisingly, intraclonal variation, which is not a feature of V(H) genes in CLL, was found in 6 of 8 cases in both subsets. These data indicate that somatic mutation of the V(H) and bcl-6 loci may not necessarily occur in tandem in CLL, suggesting diverse pathways operating on the 2 genes. PMID- 10828041 TI - Pleiotropic role of lyn kinase in leukotriene B(4)-induced eosinophil activation. AB - The authors have examined the role of the src-family of protein tyrosine kinases in leukotriene B(4) (LTB(4))-induced activation of guinea-pig eosinophils. Western blot analysis identified the src-like protein tyrosine kinases p53(lyn), p56(lyn), p56/59(hck), p55(fgr), and p56(lck) whereas p60(src), p62(yes), p55(blk), and p59(fyn) were not detected. LTB(4) promoted a rapid increase in p53/56(lyn) activity in eosinophils, which peaked at 5 seconds and remained elevated at 60 seconds; hck, fgr, and lck were not activated. A role for p53/56(lyn) in eosinophil activation was investigated with the use of the src selective inhibitor PP1 (1 micromol/L to 10 micromol/L), which attenuated LTB(4) stimulated p53/56(lyn) activity and the phosphorylation of extracellular signal regulated kinase-2 in intact cells. At comparable concentrations, PP1 was also shown to attenuate LTB(4)-induced nicotinamide adenine dinucleotide phosphate (reduced form) (NADPH) oxidase activation, chemotaxis, and Ca(++)-dependent [(3)H]arachidonic acid (AA) release. Moreover, an inhibitor of mitogen-activated protein kinase kinase-1, PD 098059, significantly inhibited LTB(4)-induced chemotaxis but had no effect on oxidant production or [(3)H]AA release. Collectively, these results implicate lyn kinase in LTB(4)-induced eosinophil activation through the recruitment of divergent cell-signaling pathways. PMID- 10828042 TI - Interferon-gamma improves splicing efficiency of CYBB gene transcripts in an interferon-responsive variant of chronic granulomatous disease due to a splice site consensus region mutation. AB - X-linked chronic granulomatous disease (CGD) derives from defects in the CYBB gene, which encodes the gp91-phox component of NADPH oxidase. We studied the molecular basis of the disease in a kindred with variant CGD, due to a single base substitution at the sixth position of CYBB first intron. The patients' phagocytes have been shown previously to greatly increase superoxide release in response to interferon-gamma (IFN-gamma) in vitro and in vivo. We examined CYBB gene expression in an Epstein-Barr virus (EBV)-transformed B-cell line from 1 patient in this kindred. These cells showed markedly decreased levels of CYBB transcripts in total RNA (5% of normal) and nuclear RNA (1.4% of normal), despite equal CYBB transcription rates in the CGD and control cells. Incubation with IFN gamma produced a 3-fold increase in CYBB total messenger RNA (mRNA) levels in the patient's cells, and decreased nuclear transcripts to undetectable levels. Reverse transcriptase-polymerase chain reaction analysis of RNA splicing revealed a preponderance of unspliced CYBB transcripts in the patient's nuclear RNA. In vitro incubation with IFN-gamma increased by 40% the ratio of spliced relative to unspliced CYBB mRNA in nuclei from the CGD B-cell line. Total RNA harvested from the same patient's monocytes, on and off therapy with IFN-gamma, showed a similar improvement in splicing. We conclude that IFN-gamma partially corrects a nuclear processing defect due to the intronic mutation in the CYBB gene in this kindred, most likely by augmentation of nuclear export of normal transcripts, and improvement in the fidelity of splicing at the first intron. PMID- 10828043 TI - Hemoglobin switching in unicellular erythroid culture of sibling erythroid burst forming units: kit ligand induces a dose-dependent fetal hemoglobin reactivation potentiated by sodium butyrate. AB - Mechanisms underlying fetal hemoglobin (HbF) reactivation in adult life have not been elucidated; particularly, the role of growth factors (GFs) is controversial. Interestingly, histone deacetylase (HD) inhibitors (sodium butyrate, NaB, trichostatin A, TSA) reactivate HbF. We developed a novel model system to investigate HbF reactivation: (1) single hematopoietic progenitor cells (HPCs) were seeded in serum-free unilineage erythroid culture; (2) the 4 daughter cells (erythroid burst-forming units, [BFU-Es]), endowed with equivalent proliferation/differentiation and HbF synthesis potential, were seeded in 4 unicellular erythroid cultures differentially treated with graded dosages of GFs and/or HD inhibitors; and (3) HbF levels were evaluated in terminal erythroblasts by assay of F cells and gamma-globin content (control levels, 2.4% and 1.8%, respectively, were close to physiologic values). HbF was moderately enhanced by interleukin-3 (IL-3) and granulocyte-macrophage colony-stimulating factor treatment (up to 5%-8% gamma-globin content), while sharply reactivated in a dose dependent fashion by c-kit ligand (KL) and NaB (20%-23%). The stimulatory effects of KL on HbF production and erythroid cell proliferation were strictly correlated. A striking increase of HbF was induced by combined addition of KL and NaB or TSA (40%-43%). This positive interaction is seemingly mediated via different mechanisms: NaB and TSA may modify the chromatin structure of the beta globin gene cluster; KL may activate the gamma-globin promoter via up-modulation of tal-1 and possibly FLKF transcription factors. These studies indicate that KL plays a key role in HbF reactivation in adult life. Furthermore, combined KL and NaB administration may be considered for sickle cell anemia and beta-thalassemia therapy. PMID- 10828044 TI - Evidence for HLA-related susceptibility for stroke in children with sickle cell disease. AB - Cerebral infarction occurs in one quarter of all children with sickle cell anemia (SCA). There is an increased risk of stroke in siblings with SCA, suggesting genetic factors may influence risk of stroke. The authors investigated whether HLA type was associated with risk of stroke in children with SCA. Fifty-three patients with SCA underwent complete HLA typing at both HLA class I (HLA-A, B) and HLA class II (HLA-DR, DQ, DP) loci. Of the 53 patients, 22 had magnetic resonance imagining (MRI)-documented evidence of cerebral infarction, and the remaining 31 patients had negative MRI scans. Comparison of the results of HLA typing between the SCA patients with a positive and those with a negative MRI documented that the 2 groups differed with respect to the class I HLA-B (P =.012), and the class II HLA-DRB1 (P =.0008) and DQB1 (P =.029). Susceptibility associations at the HLA-DRB1 locus included both DR3 alleles, where DRB1*0301 and *0302 were both associated with an increased risk of stroke. Protective associations were found in the DR2 group, where DRB1*1501 was protective for stroke. DQB1*0201, which is in linkage disequilibrium with DRB1*0301, was also associated with stroke. Similarly, DQB1*0602, in linkage disequilibrium with DRB1*1501, was protective. Specific HLA alleles may influence the risk of stroke in children with SCA. HLA typing may prove useful in identifying SCA patients at higher risk for stroke. PMID- 10828045 TI - Analysis of ferrochelatase expression during hematopoietic development of embryonic stem cells. AB - Ferrochelatase, the last enzyme in the heme pathway, chelates protoporphyrin IX and iron to form heme and is mutated in protoporphyria. The ferrochelatase gene is expressed in all tissues at low levels to provide heme for essential heme containing proteins and is up-regulated during erythropoiesis for the synthesis of hemoglobin. The human ferrochelatase promoter contains 2 Sp1 cis-elements and GATA and NF-E2 sites, all of which bind their cognate trans-acting factors in vitro. To investigate the role of these elements during erythropoiesis, we introduced expression of the green fluorescent protein (EGFP) transgenes driven by various ferrochelatase promoter fragments into a single locus in mouse embryonic stem cells. EGFP expression was monitored during hematopoietic differentiation in vitro using flow cytometry. We show that a promoter fragment containing the Sp1 sites, the NF-E2 and GATA elements, was sufficient to confer developmental-specific expression of the EGFP transgene, with an expression profile identical to that of the endogenous gene. In this system the -0.275 kb NF E2 cis-element is required for erythroid-enhanced expression, the GATA cis element functions as a stage-specific repressor and enhancer, and elements located between -0.375kb and -1.1kb are necessary for optimal levels of expression. Ferrochelatase mRNA increased before the primitive erythroid-cell stage without a concomitant increase in ferrochelatase protein, suggesting the presence of a translational control mechanism. Because of the sensitivity of this system, we were able to assess the effect of an A-to-G polymorphism identified in the promoters of patients with protoporphyria. There was no effect of the G haplotype on transcriptional activity of the -1.1 kb transgene. PMID- 10828046 TI - FKLF-2: a novel Kruppel-like transcriptional factor that activates globin and other erythroid lineage genes. AB - FKLF-2, a novel Kruppel-type zinc finger protein, was cloned from murine yolk sac. The deduced polypeptide sequence of 289 amino acids has 3 contiguous zinc fingers at the near carboxyl-terminal end, an amino-terminal domain characterized by its high content of alanine and proline residues and a carboxyl-terminal domain rich in serine residues. By Northern blot hybridization, the human homologue of FKLF-2 is expressed in the bone marrow and striated muscles and not in 12 other human tissues analyzed. FKLF-2 is constitutively expressed in established cell lines with an erythroid phenotype, but it is inconsistently expressed in cell lines with myeloid or lymphoid phenotypes. The expression of FKLF-2 messenger RNA (mRNA) is up-regulated after induction of mouse erythroleukemia cells. In luciferase assays, FKLF-2 activates predominantly the gamma, and to a lesser degree, the epsilon and beta globin gene promoters. The activation of gamma gene promoter does not depend on the presence of an HS2 enhancer. FKLF-2 activates the gamma promoter predominantly by interacting with the gamma CACCC box, and to a lesser degree through interaction with the TATA box or its surrounding DNA sequences. FKLF-2 also activated all the other erythroid specific promoters we tested (GATA-1, glycophorin B, ferrochelatase, porphobilinogen deaminase, and 5-aminolevulinate synthase). These results suggest that in addition to globin, FKLF-2 may be involved in activation of transcription of a wide range of genes in the cells of the erythroid lineage. PMID- 10828047 TI - Estimating the prevalence of pyruvate kinase deficiency from the gene frequency in the general white population. AB - Pyruvate kinase (PK) deficiency is the most common cause of hereditary nonspherocytic hemolytic anemia. The prevalence of this deficiency is unknown, though some estimates have been made based on the frequency of low red cell PK activity in the population. An additional 20 patients with hereditary nonspherocytic hemolytic anemia caused by PK deficiency have been genotyped. One previously unreported mutation 1153C-->T (R385W) was encountered. The relative frequency of PK mutations in patients with hemolytic anemia caused by PK deficiency was calculated from the 18 white patients reported here and from 102 patients previously reported in the literature. DNA samples from 3785 subjects from different ethnic groups have been screened for the 4 more frequently encountered mutations-c.1456 C-->T(1456T), c.1468 C-->T(1468T), c.1484 C- >T(1484T), and c.1529 G6A (1529A)-by allele-specific oligonucleotide hybridization. Among white patients the frequency of the 1456T mutation was 3.50 x 10(-3); that of the 1529A mutation was 2.03 x 10(-3). Among African Americans the frequency of the 1456T mutation was 3.90 x 10(-3) The only mutation found in the limited number of Asians tested was 1468T at a frequency of 7.94 x 10(-3). Based on the gene frequency of the 1529A mutation in the white population and on its relative abundance in patients with hemolytic anemia caused by PK deficiency, the prevalence of PK deficiency is estimated at 51 cases per million white population. This number would be increased by inbreeding and decreased by failure of patients with PK deficiency to survive. PMID- 10828048 TI - Acquired DNA mutations associated with in vivo hydroxyurea exposure. AB - Hydroxyurea (HU) is an effective therapeutic agent for patients with myeloproliferative disorders (MPDs) or sickle cell disease (SCD). Short-term HU toxicities primarily include transient myelosuppression, but long-term HU risks have not been defined. The mutagenic and carcinogenic potential of HU is not established, although HU has been associated with an increased risk of leukemia in some patients with MPD. In this study, 2 assays were used to quantitate acquired somatic DNA mutations in peripheral blood mononuclear cells (PBMCs) after in vivo HU exposure. The HPRT assay measures hypoxanthine phosphoribosyl transferase (hprt) mutations, while the VDJ assay identifies "illegitimate" T cell receptor Vgamma-Jbeta interlocus recombination events. PBMCs were analyzed from patients with MPD, adults and children with SCD, and normal controls. MPD patients with prolonged HU exposure had numbers of DNA mutations equivalent to patients with low HU exposure or controls. Similarly, adults with SCD had equivalent numbers of DNA mutations regardless of HU exposure. Children with SCD and 30-month HU exposure had equivalent hprt(-) mutations but significantly more VDJ mutations (1.82 +/- 1.20 events per microg DNA) than children with 7-month HU exposure (1.58 +/- 0.87 events) or no HU exposure (1.06 +/- 0.45 events), P =.04 by analysis of variance. Taken together, these data suggest that the mutagenic and carcinogenic potential of in vivo HU therapy is low. Although increased numbers of illegitimate VDJ recombination events do not directly portend leukemia, young patients with SCD and HU exposure should be monitored serially for increases in DNA mutations. PMID- 10828049 TI - The duffy-binding-like domain 1 of Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) is a heparan sulfate ligand that requires 12 mers for binding. AB - The Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1), present on the surfaces of parasitized red blood cells (pRBC), mediates rosetting, a virulent phenotype. Here, we show that pRBC specifically bind heparan sulfate (HS) and heparin onto their surfaces and that the rosetting ligand PfEMP1 specifically adheres to heparin-Sepharose when extracted from the surfaces of radioiodinated infected RBC. An analysis of the binding properties of the different regions of PfEMP1 provides evidence that the Duffy-binding-like domain-1 (DBL-1) is the predominant ligand involved in HS and heparin binding. Soluble DBL-1 requires a minimal heparin fragment size of a 12-mer ( approximately 4 kd) for binding and is critically dependent on N-sulfation. A 12-mer is also the minimal heparin fragment that disrupts naturally formed rosettes. DBL-1 binds specifically to erythrocytes and also to HS from endothelial cells and human aorta but not to chondroitin sulfate A, suggesting that different PfEMP1s mediate adhesion to distinct glycosaminoglycans in individual malaria parasites. Present data suggest that HS on endothelial cells may also be involved in the sequestration of pRBC. Elucidation of these binding mechanisms opens up new possibilities for therapeutic strategies targeting adhesive interactions of pRBC. PMID- 10828050 TI - Independent formation of DnaseI hypersensitive sites in the murine beta-globin locus control region. AB - Mammalian beta-globin loci are composed of multiple orthologous genes whose expression is erythroid specific and developmentally regulated. The expression of these genes both from the endogenous locus and from transgenes is strongly influenced by a linked 15-kilobase region of clustered DNaseI hypersensitive sites (HSs) known as the locus control region (LCR). The LCR encompasses 5 major HSs, each of which is highly homologous among humans, mice, and other mammals. To analyze the function of individual HSs in the endogenous murine beta-globin LCR, we have used homologous recombination in embryonic stem cells to produce 5 mouse lines, each of which is deficient for 1 of these major HSs. In this report, we demonstrate that deletion of the conserved region of 5'HS 1, 2, 3, 4, or 5/6 abolishes HS formation at the deletion site but has no influence on the formation of the remaining HSs in the LCR. Therefore, in the endogenous murine locus, there is no dominant or initiating site whose formation must precede the formation of the other HSs. This is consistent with the idea that HSs form autonomously. We discuss the implications of these findings for current models of beta-globin regulation. PMID- 10828051 TI - Effect of leukocyte compatibility on neutrophil increment after transfusion of granulocyte colony-stimulating factor-mobilized prophylactic granulocyte transfusions and on clinical outcomes after stem cell transplantation. AB - The primary limitations of granulocyte transfusions include low component cell dose and leukocyte incompatibility. Component cell dose improved with granulocyte colony-stimulating factor (G-CSF) mobilization, and the transfusion of G-CSF mobilized, human leukocyte antigen (HLA)-matched granulocyte components resulted in significant, sustained absolute neutrophil count (ANC) increments. However, the effect of leukocyte compatibility on outcomes with G-CSF-mobilized granulocyte transfusions is unclear. The objectives were to determine the effect of leukocyte compatibility on ANC increments and selected clinical outcomes after transfusion of prophylactic, G-CSF-mobilized granulocyte components into neutropenic recipients of autologous peripheral blood stem cell (PBSC) transplants. Beginning on transplant day 2, 23 evaluable recipients were scheduled to receive 4 alternate-day transfusions of granulocyte components apheresed from a single donor given G-CSF. G-CSF was also given to recipients after transplantation. Recipient ANC was determined before and sequentially after each granulocyte transfusion to determine the peak ANC increment. Leukocyte compatibility was determined at study entry only by a lymphocytotoxicity screening assay (s-LCA) against a panel of HLA-defined cells. Eight recipients had positive s-LCA. On days 2 and 4, the mean peak ANC increments after granulocyte transfusion were comparable between the cohorts with positive and negative s-LCA. However, the mean peak ANC increments on day 6 (246/microL vs 724/microL; P =.05) and day 8 (283/microL vs 1079/microL; P =.06) were lower in the cohort with positive s-LCA, in spite of the transfusion of comparable component cell doses. Adverse reactions occurred with only 5 of 87 (5.7%) granulocyte transfusions and were not associated with leukocyte compatibility test results. Platelet increments, determined 1 hour after granulocyte transfusion, were comparable between the cohorts. Although the 2 cohorts received PBSC components with similar CD34(+) cell doses, the cohort with a positive s-LCA had delayed neutrophil engraftment and a greater number of febrile days and required more days of intravenous antibiotics and platelet transfusions. Leukocyte incompatibility adversely affected ANC increments after the transfusion of G-CSF-mobilized granulocyte components and clinical outcomes after PBSC transplantation. PMID- 10828052 TI - Induction of oral tolerance in splenocyte recipients toward pretransplant antigens ameliorates chronic graft versus host disease in a murine model. AB - Chronic graft versus host disease (cGVHD) is a major complication that can develop after bone marrow transplantation. It involves an immune-mediated attack by transplanted donor lymphocytes, and often results in inflammatory damage of host target organs. Immune hyporesponsiveness induced by oral antigen administration has been recently shown to prevent the development of cGVHD in a murine model. The aim of this study was to evaluate whether tolerance induction in bone marrow transplant (BMT) recipients after transplantation, toward their pretransplant antigens, can alleviate preexisting cGVHD in a mouse model. cGVHD was generated by infusing 2.5 x 10(7) splenocytes from B10.D2 donor mice, to sublethally irradiated (6 Gy) BALB/c recipient mice, which differ by minor histocompatibility antigens. Transplantation resulted in cGVHD, with characteristic scleroderma-like cutaneous fibrosis, increased skin collagen content, decreased body weight, and hepatic and small bowel inflammation. Oral tolerance was induced by feeding recipient BALB/c mice with proteins extracted from BALB/c splenocytes for 11 days after B10.D2 splenocyte transplantation. Tolerance induction was evidenced by a significant reduction in mixed lymphocyte response of effector splenocytes from tolerant BALB/c mice transplanted with B10.D2 splenocytes against BALB/c target splenocytes. Oral tolerance decreased skin collagen deposits. Reduction of collagen alpha1(I) gene expression and skin collagen were shown by in situ hybridization and histochemistry, respectively. Liver and bowel biopsy specimens revealed less inflammation. Serum IL-10 levels were higher in tolerant mice than in controls, whereas IFNgamma was significantly reduced. Oral tolerance of BMT recipients toward their pretransplant antigens after splenocyte transplantation down-regulated the immune attack by transplanted cells, thus ameliorating cGVHD. PMID- 10828053 TI - Cotransplantation of human stromal cell progenitors into preimmune fetal sheep results in early appearance of human donor cells in circulation and boosts cell levels in bone marrow at later time points after transplantation. AB - Both in utero and postnatal hematopoietic stem cell (HSC) transplantation would benefit from the development of approaches that produce increased levels of engraftment or a reduction in the period of time required for reconstitution. We used the in utero model of human-sheep HSC transplantation to investigate ways of improving engraftment and differentiation of donor cells after transplantation. We hypothesized that providing a more suitable microenvironment in the form of human stromal cell progenitors simultaneously with the transplanted human HSC would result in higher rates of engraftment or differentiation of the human cells in this xenogeneic model. The results presented here demonstrate that the cotransplantation of both autologous and allogeneic human bone marrow-derived stromal cell progenitors resulted in an enhancement of long-term engraftment of human cells in the bone marrow of the chimeric animals and in earlier and higher levels of donor cells in circulation both during gestation and after birth. By using marked stromal cells, we have also demonstrated that injected stromal cells alone engraft and remain functional within the sheep hematopoietic microenvironment. Application of this method to clinical HSC transplantation could potentially lead to increased levels of long-term engraftment, a reduction in the time for hematopoietic reconstitution, and a means of delivery of foreign genes to the hematopoietic system. PMID- 10828054 TI - Expression of CD94/NKG2A and killer immunoglobulin-like receptors in NK cells and a subset of extranodal cytotoxic T-cell lymphomas. AB - Thirty-two natural killer (NK) and cytotoxic T-cell lymphomas and 14 noncytotoxic nodal T-cell lymphoma controls were immunostained with the use of monoclonal antibodies reactive against NK-cell receptor (NKR) molecules (CD94, NKG2A, p58.2, p58.1, p140, p70, p50.3). All NK-cell lymphomas (4 nasal/oral and 1 intestinal) expressed at least 1 NKR, the CD94/NKG2A complex. Two were positive for 1 or more killer immunoglobulin-like receptors. Of 15 extranodal cytotoxic T-cell lymphomas, 3 expressed CD94, including 2 intestinal and 1 hepatosplenic gammadelta T-cell lymphomas. In contrast, none of the nodal lymphomas were positive. Detection of NKRs may provide a useful tool to confirm the diagnosis of NK-cell lymphomas and to delineate a subgroup of cytotoxic T-cell lymphomas. Expression of NKRs only in extranodal cytotoxic T-cell lymphomas might reflect differences in the homing capabilities of cytotoxic T cells expressing NKRs in normal individuals and might be influenced in part by localized chronic immune reactions. PMID- 10828055 TI - Reduction of lysosomal storage in murine mucopolysaccharidosis type VII by transplantation of normal and genetically modified macrophages. AB - This study examined the ability of macrophages to serve as target cells of gene therapy for mucopolysaccharidosis (MPS) type VII using a murine model. Bone marrow cells were harvested from syngeneic normal mice and differentiated to macrophages. These cells were given to nonmyeloablated MPS VII mice. After transplantation, donor cells populated the liver and spleen. The pathologic improvement at day 38 after transplantation was significant and glycosaminoglycan storage was reduced. To develop gene therapy using this system, a retroviral vector expressing human beta-glucuronidase (HBG) was used to infect macrophages cultivated from MPS VII mice and given to nonmyeloablated MPS VII mice. At 38 days after transplantation, HBG-positive cells were still observed histochemically and pathologic improvement was significant. These observations suggest that macrophage transplantation is a promising method for treatment of murine MPS VII without myeloablation, and macrophages may be good target cells for ex vivo gene therapy for MPS VII. PMID- 10828056 TI - Structural insights into the protein splicing mechanism of PI-SceI. AB - PI-SceI is a member of a class of proteins (inteins) that excise themselves from a precursor protein and in the process ligate the flanking protein sequences (exteins). We report here the 2.1-A resolution crystal structure of a PI-SceI miniprecursor (VMA29) containing 10 N-terminal extein residues and 4 C-terminal extein residues. Mutations at the N- and C-terminal splicing junctions, blocking in vivo protein splicing, allowed the miniprecursor to be purified and crystallized. The structure reveals both the N- and C-terminal scissile peptide bonds to be in distorted trans conformations (tau approximately 100 degrees ). Modeling of the wild-type PI-SceI based on the VMA29 structure indicates a large conformational change (movement of >9 A) must occur to allow transesterification to be completed. A zinc atom was discovered at the C-terminal splicing junction. Residues Cys(455), His(453), and Glu(80) along with a water molecule (Wat(53)) chelate the zinc atom. The crystal structure of VMA29 has captured the intein in its pre-spliced state. PMID- 10828057 TI - Specific interaction of TAFII105 with OCA-B is involved in activation of octamer dependent transcription. AB - TAF(II)105 is a TFIID-associated factor highly expressed in B lymphocytes. This subunit is found in a small portion of TFIID complexes and is homologous to human TAF(II)130 and Drosophila TAF(II)110. In the present study we show that TAF(II)105 is involved in transcription activation directed by the B cell specific octamer element found in many B cell-specific genes. B cells overexpressing TAF(II)105 display higher octamer-dependent transcription, whereas expression of a C-terminal truncated form of TAF(II)105 inhibits octamer transcription in a dominant negative manner. In addition, antibodies directed against TAF(II)105 specifically inhibit octamer-dependent transcription. Reporter gene analysis revealed that TAF(II)105 elevates octamer transcription in the presence of OCA-B, a cofactor subunit of Oct1 and Oct2 proteins. In vitro binding assays and functional studies established that the effect of TAF(II)105 on octamer activity involves interaction of TAF(II)105 with octamer-binding complexes via the C-terminal activation domain of OCA-B. These findings link TAF(II)105 coactivator function to B cell-specific transcription. PMID- 10828058 TI - Sphingomyelinase activity associated with human plasma low density lipoprotein. AB - Isolated human plasma low density lipoprotein (LDL) was observed to possess sphingomyelinase activity. Accordingly, the formation of ceramide was catalyzed by LDL at 37 degrees C using tertiary liposomes composed of sphingomyelin (mole fraction (x) = 0.2), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (x = 0.7), 1, 2-dimyristoyl-sn-glycero-3-phospho-rac-glycerol (x = 0.1), and either the fluorescent sphingomyelin analog Bodipy-sphingomyelin or [(14)C]sphingomyelin as substrates. However, this activity was not present in either very low density lipoprotein or the high density lipoprotein subfractions HDL(2) and HDL(3). Oxidation of LDL abrogated its sphingomyelinase activity. Aggregation of the liposomes upon incubation with LDL was evident from the light scattering measurements. Microinjection of LDL to the surface of giant liposomes composed of 1-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine (SOPC), N-palmitoyl-d sphingomyelin (C16:0-sphingomyelin), and Bodipy-sphingomyelin as a fluorescent tracer (0.75:- 0.20:0.05, respectively) revealed the induction of vectorial budding of vesicles, resembling endocytosis. PMID- 10828059 TI - ERK2 mitogen-activated protein kinase binding, phosphorylation, and regulation of the PDE4D cAMP-specific phosphodiesterases. The involvement of COOH-terminal docking sites and NH2-terminal UCR regions. AB - The cAMP-specific phosphodiesterase family 4, subfamily D, isoform 3 (PDE4D3) is shown to have FQF and KIM docking sites for extracellular signal-regulated kinase 2 (ERK2) (p42(MAPK)). These straddle the target residue, Ser(579), for ERK2 phosphorylation of PDE4D3. Mutation of either or both of these docking sites prevented ERK2 from being co-immunoprecipitated with PDE4D3, ablated the ability of epidermal growth factor to inhibit PDE4D3 through ERK2 action in transfected COS cells, and attenuated the ability of ERK2 to phosphorylate PDE4D3 in vitro. The two conserved NH(2)-terminal blocks of sequence, called upstream conserved regions 1 and 2 (UCR1 and UCR2), that characterize PDE4 long isoforms, are proposed to amplify the small, inherent inhibitory effect that ERK2 phosphorylation exerts on the PDE4D catalytic unit. In contrast to this, the lone intact UCR2 region found in PDE4D1 directs COOH-terminal ERK2 phosphorylation to cause the activation of this short isoform. From the analysis of PDE4D3 truncates, it is suggested that UCR1 and UCR2 provide a regulatory signal integration module that serves to orchestrate the functional consequences of ERK2 phosphorylation. The PDE4D gene thus encodes a series of isoenzymes that are either inhibited or activated by ERK2 phosphorylation and thereby offers the potential for ERK2 activation either to increase or decrease cAMP levels in cellular compartments. PMID- 10828060 TI - Peroxisome proliferator-activated receptor alpha activators improve insulin sensitivity and reduce adiposity. AB - Fibrates and glitazones are two classes of drugs currently used in the treatment of dyslipidemia and insulin resistance (IR), respectively. Whereas glitazones are insulin sensitizers acting via activation of the peroxisome proliferator activated receptor (PPAR) gamma subtype, fibrates exert their lipid-lowering activity via PPARalpha. To determine whether PPARalpha activators also improve insulin sensitivity, we measured the capacity of three PPARalpha-selective agonists, fenofibrate, ciprofibrate, and the new compound GW9578, in two rodent models of high fat diet-induced (C57BL/6 mice) or genetic (obese Zucker rats) IR. At doses yielding serum concentrations shown to activate selectively PPARalpha, these compounds markedly lowered hyperinsulinemia and, when present, hyperglycemia in both animal models. This effect relied on the improvement of insulin action on glucose utilization, as indicated by a lower insulin peak in response to intraperitoneal glucose in ciprofibrate-treated IR obese Zucker rats. In addition, fenofibrate treatment prevented high fat diet-induced increase of body weight and adipose tissue mass without influencing caloric intake. The specificity for PPARalpha activation in vivo was demonstrated by marked alterations in the expression of PPARalpha target genes, whereas PPARgamma target gene mRNA levels did not change in treated animals. These results indicate that compounds with a selective PPARalpha activation profile reduce insulin resistance without having adverse effects on body weight and adipose tissue mass in animal models of IR. PMID- 10828061 TI - Characterization of Cdk2-cyclin E complexes in plasma membrane and endosomes of liver parenchyma. Insulin-dependent regulation. AB - Rat liver parenchyma Golgi/endosomes fractions harbor a tyrosine-phosphorylated 34-kDa protein. Screening of Golgi, endosomes (ENs), plasmalemma (PM), and cytosolic (Cyt) fractions revealed the presence of the mitotic kinase Cdk2 in ENs, PM, and Cyt. The fluid phase endocytic marker horseradish peroxidase gained access to the endosomal Cdk2, confirming its localization. Cdk2 was shown to be associated to cyclin E and was active in ENs and PM fractions. The administration of a single dose of insulin (1.5 microgram/100 g, body weight) induced a time dependent activation of the insulin receptor kinase in these structures. Insulin receptor-kinase activation was followed by the inhibition of immunoprecipitated Cdk2-cyclin E kinase activity in PM and the progressive disappearance of cyclin E. In marked contrast, no such effect was observed in ENs. The injection of a phosphotyrosyl phosphatase inhibitor (bpV(phen)) increased the levels of cyclin E in ENs and PM. A massive recruitment of p27(kip1) was observed in the Cdk2-cyclin E complexes isolated from PM and Cyt but not from ENs. In vitro, Cdk2-cyclin E complexes have the capacity to inhibit the formation of hybrid structures containing horseradish peroxidase and radioiodinated epidermal growth factor. Therefore, in the PM and ENs of adult rat liver, an active and regulated pool of the mitotic kinase Cdk2-cyclin E and some yet to be defined effectors are present. Cdk2 may contribute to the modulation of transport events and/or maintenance of the topology of endocytic elements. PMID- 10828062 TI - Induction of platelet-derived growth factor B-chain expression by transforming growth factor-beta involves transactivation by Smads. AB - Transforming growth factor-beta (TGF-beta) regulates a diverse array of biological processes, such as proliferation, differentiation, extracellular matrix production, and apoptosis. In cultured vascular endothelial cells, TGF beta induces the expression of platelet-derived growth factor (PDGF) B-chain, a mitogen and chemoattractant, at the level of transcription. The molecular mechanism(s) underlying this process are not presently understood. In this study, we performed serial 5' deletion and transient transfection analysis to define a region in the PDGF-B promoter mediating inducible responsiveness to TGF-beta. This region contains an atypical nucleotide recognition element for the Smad family of transcriptional regulators. Electrophoretic mobility shift analysis revealed that nuclear proteins bound to this site in a transient and specific manner. Supershift studies demonstrated the physical association of Smad4 with the promoter. Overexpression of Smad4 activated the PDGF-B promoter and superinduced PDGF-B promoter-dependent expression in cells exposed to TGF-beta. Moreover, simultaneous cotransfection of Smad3 and Smad4 activated the PDGF-B promoter. This effect was attenuated when Smad4 was substituted with its dominant negative counterpart. Mutation of the (-81)CAGA(-78) motif in the PDGF-B promoter abrogated Smad-inducible promoter-dependent expression. Overexpression of Smad2 and Smad3 transactivated the PDGF-B promoter in a synergistic manner. These findings demonstrate the existence of a novel, functional binding element in the proximal region of the PDGF-B promoter mediating responsiveness to TGF-beta. PMID- 10828063 TI - Signaling across the platelet adhesion receptor glycoprotein Ib-IX induces alpha IIbbeta 3 activation both in platelets and a transfected Chinese hamster ovary cell system. AB - In platelets, alpha(IIb)beta(3) exists in a form that cannot bind adhesive proteins in the plasma; although it can interact with immobilized fibrinogen it cannot interact with immobilized von Willebrand factor in the vessel wall. Soluble agonists such as thrombin convert alpha(IIb)beta(3) to a form that recognizes soluble and immobilized ligands. Attempts to reconstitute alpha(IIb)beta(3) activation in a non-hematopoietic, nucleated cell system have been unsuccessful. In the present study, we have developed a transfected Chinese hamster ovary cell model in which alpha(IIb)beta(3) activation is induced by signaling across glycoprotein (GP) Ib-IX by its ligand, von Willebrand factor. GPIb-IX activates not only the transfected alpha(IIb)beta(3) but also endogenous alpha(v)beta(3). Activation of the pathways leading to integrin activation occurred even in cells transfected with GPIb-IX lacking the domain on GPIbalpha that binds 14-3-3 or that which binds actin-binding protein. These studies demonstrate that signals induced by interaction of GPIb-IX with von Willebrand factor lead to alpha(IIb)beta(3) activation and suggest that the signaling pathways by which GPIb-IX induces alpha(IIb)beta(3) activation are different to those used by thrombin. Elucidation of these differences may provide insights into therapeutic ways in which to inhibit integrin activation in selective clinical settings. PMID- 10828064 TI - WNK1, a novel mammalian serine/threonine protein kinase lacking the catalytic lysine in subdomain II. AB - We have cloned and characterized a novel mammalian serine/threonine protein kinase WNK1 (with no lysine (K)) from a rat brain cDNA library. WNK1 has 2126 amino acids and can be detected as a protein of approximately 230 kDa in various cell lines and rat tissues. WNK1 contains a small N-terminal domain followed by the kinase domain and a long C-terminal tail. The WNK1 kinase domain has the greatest similarity to the MEKK protein kinase family. However, overexpression of WNK1 in HEK293 cells exerts no detectable effect on the activity of known, co transfected mitogen-activated protein kinases, suggesting that it belongs to a distinct pathway. WNK1 phosphorylates the exogenous substrate myelin basic protein as well as itself mostly on serine residues, confirming that it is a serine/threonine protein kinase. The demonstration of activity was striking because WNK1, and its homologs in other organisms lack the invariant catalytic lysine in subdomain II of protein kinases that is crucial for binding to ATP. A model of WNK1 using the structure of cAMP-dependent protein kinase suggests that lysine 233 in kinase subdomain I may provide this function. Mutation of this lysine residue to methionine eliminates WNK1 activity, consistent with the conclusion that it is required for catalysis. This distinct organization of catalytic residues indicates that WNK1 belongs to a novel family of serine/threonine protein kinases. PMID- 10828065 TI - Characterization of MyD118, Gadd45, and proliferating cell nuclear antigen (PCNA) interacting domains. PCNA impedes MyD118 AND Gadd45-mediated negative growth control. AB - MyD118 and Gadd45 are related genes encoding for proteins that play important roles in negative growth control, including growth suppression and apoptosis. MyD118 and Gadd45 are related proteins that previously were shown to interact with proliferating cell nuclear antigen (PCNA), implicated in DNA replication, DNA repair, and cell cycle progression. To establish the role of MyD118 and Gadd45 interactions with PCNA, in this work we sought to identify the interacting domains and analyze the significance of this interaction in negative growth control. Using complementary in vivo and in vitro interaction assays the N terminal (1-46) and middle (100-127) regions of PCNA were identified as harboring MyD118- and Gadd45 interacting domains, whereas PCNA interacting domains within MyD118 and Gadd45 were localized to the C termini of these proteins (amino acids 114-156 and 137-165, respectively). These findings provide first evidence that similar domains within MyD118 and Gadd45 mediate interactions with PCNA. Importantly, ectopic expression of MyD118 or Gadd45 N-terminal peptides, lacking the PCNA interacting domain, was found to suppress colony formation or induce apoptosis more efficiently than the full-length proteins. These findings suggest that interaction of MyD118 or Gadd45 with PCNA, in essence, serves to impede negative growth control. PMID- 10828066 TI - Identification and characterization of human mitochondrial tryptophanyl-tRNA synthetase. AB - A full-length cDNA clone encoding the human mitochondrial tryptophanyl-tRNA synthetase (h(mt)TrpRS) has been identified. The deduced amino acid sequence shows high homology to both the mitochondrial tryptophanyl-tRNA synthetase ((mt)TrpRS) from Saccharomyces cerevisiae and to different eubacterial forms of tryptophanyl-tRNA synthetase (TrpRS). Using the baculovirus expression system, we have expressed and purified the protein with a carboxyl-terminal histidine tag. The purified His-tagged h(mt)TrpRS catalyzes Trp-dependent exchange of PP(i) in the PP(i)-ATP exchange assay. Expression of h(mt)TrpRS in both human and insect cells leads to high levels of h(mt)TrpRS localizing to the mitochondria, and in insect cells the first 18 amino acids constitute the mitochondrial localization signal sequence. Until now the human cytoplasmic tryptophanyl-tRNA synthetase (hTrpRS) was thought to function as the h(mt)TrpRS, possibly in the form of a splice variant. However, no mitochondrial localization signal sequence was ever detected and the present identification of a different (mt)TrpRS almost certainly rules out that possibility. The h(mt)TrpRS shows kinetic properties similar to human mitochondrial phenylalanyl-tRNA synthetase (h(mt)PheRS), and h(mt)TrpRS is not induced by interferon-gamma as is hTrpRS. PMID- 10828067 TI - Interaction of GRASP, a protein encoded by a novel retinoic acid-induced gene, with members of the cytohesin family of guanine nucleotide exchange factors. AB - A novel, retinoic acid-induced gene, GRP1-associated scaffold protein (GRASP), was isolated from P19 embryonal carcinoma cells using a subtractive screening strategy. GRASP was found to be highly expressed in brain and exhibited lower levels of expression in lung, heart, embryo, kidney, and ovary. The predicted amino acid sequence of GRASP is characterized by several putative protein-protein interaction motifs, suggesting that GRASP may be a component of a larger protein complex in the cell. Although GRASP does not harbor a predicted membrane spanning domain(s), the protein was observed to be associated with the plasma membrane of transiently transfected mammalian cells. Yeast two-hybrid screening revealed that GRASP interacted strongly with the General Receptor for Phosphoinositides 1 (GRP1), a brefeldin A-insensitive guanine nucleotide exchange factor for the ADP ribosylation factor family of proteins. GRASP. GRP1 interactions were also demonstrated in vitro and in mammalian cells in which GRASP was shown to enhance GRP1 association with the plasma membrane. Furthermore, GRASP colocalized with endogenous ADP-ribosylation factors at the plasma membrane in transfected cells, suggesting that GRASP may modulate signaling by this family of small GTPases. PMID- 10828068 TI - Transient induction of ENC-1, a Kelch-related actin-binding protein, is required for adipocyte differentiation. AB - In an attempt to study molecules that play a regulatory role early in adipocyte differentiation, we identified by differential display ENC-1, a Drosophila kelch related protein. ENC-1 colocalizes with actin filaments. ENC-1 is expressed in adipose tissue, specifically in the adipose-derived stroma-vascular fraction. ENC 1 mRNA levels are transiently increased 8-12-fold early in in vitro adipocyte differentiation of primary cells of the adipose-derived stroma-vascular fraction and of 3T3-L1 cells. Treatment with the adipogenic inducers dexamethasone and methylisobutylxanthine causes an increase in ENC-1 mRNA levels specifically in preadipocytes, and methylisobutylxanthine is the main effector of ENC-1 expression. The induction of ENC-1 precedes expression of the transcription factors, peroxisome proliferator-activated receptor (PPARgamma) and CCAAT/enhancer-binding protein (C/EBPalpha), and other adipocyte markers including adipocyte fatty acid-binding protein. The ENC-1 induction correlates with the subsequent differentiation of primary stroma-vascular cells into adipocytes. Furthermore, decreasing the endogenous ENC-1 levels by stable antisense transfection, thereby preventing the transient induction, effectively inhibits 3T3-L1 adipocyte differentiation. Overall, these studies indicate that ENC-1, an actin-binding protein, plays a regulatory role early in adipocyte differentiation when cytoskeletal reorganization and cell shape change from fibroblastic preadipocytes to spherical adipocytes occur. PMID- 10828069 TI - Folding pathway mediated by an intramolecular chaperone. The inhibitory and chaperone functions of the subtilisin propeptide are not obligatorily linked. AB - The subtilisin propeptide functions as an intramolecular chaperone (IMC) that facilitates correct folding of the catalytic domain while acting like a competitive inhibitor of proteolytic activity. Upon completion of folding, subtilisin initiates IMC degradation to complete precursor maturation. Existing data suggest that the chaperone and inhibitory functions of the subtilisin IMC domain are interdependent during folding. Based on x-ray structure of the IMC subtilisin complex, we introduce a point mutation (E112A) to disrupt three hydrogen bonds that stabilize the interface between the protease and its IMC domain. This mutation within subtilisin does not alter the folding kinetics but dramatically slows down autoprocessing of the IMC domain. Inhibition of E112A subtilisin activity by the IMC added in trans is 35-fold weaker than wild-type subtilisin. Although the IMC domain displays substantial loss of inhibitory function, its ability to chaperone E112A-subtilisin folding remains intact. Our results show that (i) the chaperone activity of the IMC domain is not obligatorily linked with its ability to bind with and inhibit active subtilisin; (ii) degradation and not autoprocessing of the IMC domain is the rate-limiting step in precursor maturation; and (iii) the Glu(112) residue within the IMC subtilisin interface is not crucial for initiating folding but is important in maintaining the IMC structure capable of binding subtilisin. PMID- 10828070 TI - Regulation of neuronal cell adhesion molecule expression by NF-kappa B. AB - The neuronal cell adhesion molecule (NCAM) is a key mediator of structural plasticity in the central nervous system, but the mechanisms that control its expression are unknown. Equally, although the transcription factor NF-kappaB is present in the brain, few NF-kappaB-regulated genes relevant for central nervous system function have been identified. We have previously demonstrated that NF kappaB is activated in neuronal cultures treated with kainic acid or nitric oxide. We show here that kainic acid or nitric oxide also increase the levels of NCAM mRNA and protein in neurons and that this induction of NCAM expression is sensitive to dexamethasone and to antisense, but not missense, oligonucleotides designed to suppress NF-kappaB synthesis. Nitric oxide also stimulates protein binding to an NF-kappaB site in the promoter of the NCAM gene. This indicates that NF-kappaB, which has recently been implicated in synaptic plasticity and also in the etiology of neurodegenerative disease, plays a crucial role in the activity-dependent regulation of NCAM gene expression. In addition, since both NCAM and NF-kappaB are present in the post-synaptic density, this represents a route allowing direct communication between the synapse and the nucleus. PMID- 10828071 TI - Generating a high affinity scorpion toxin receptor in KcsA-Kv1.3 chimeric potassium channels. AB - The crystal structure of the bacterial K(+) channel, KcsA (Doyle, D. A., Morais, C. J., Pfuetzner, R. A., Kuo, A., Gulbis, J. M., Cohen, S. L., Chait, B. T., and MacKinnon, R. (1998) Science 280, 69-77), and subsequent mutagenesis have revealed a high structural conservation from bacteria to human (MacKinnon, R., Cohen, S. L., Kuo, A., Lee, A., and Chait, B. T. (1998) Science 280, 106-109). We have explored this conservation by swapping subregions of the M1-M2 linker of KcsA with those of the S5-S6 linker of the human Kv-channel Kv1.3. The chimeric K(+) channel constructs were expressed in Escherichia coli, and their multimeric state was analyzed after purification. We used two scorpion toxins, kaliotoxin and hongotoxin 1, which bind specifically to Kv1.3, to analyze the pharmacological properties of the KcsA-Kv1.3 chimeras. The results demonstrate that the high affinity scorpion toxin receptor of Kv1.3 could be transferred to KcsA. Our biochemical studies with purified KcsA-Kv1.3 chimeras provide direct chemical evidence that a tetrameric channel structure is necessary for forming a functional scorpion toxin receptor. We have obtained KcsA-Kv1.3 chimeras with kaliotoxin affinities (IC(50) values of approximately 4 pm) like native Kv1.3 channels. Furthermore, we show that a subregion of the S5-S6 linker may be an important determinant of the pharmacological profile of K(+) channels. Using available structural information on KcsA and kaliotoxin, we have developed a structural model for the complex between KcsA-Kv1.3 chimeras and kaliotoxin to aid future pharmacological studies of K(+) channels. PMID- 10828072 TI - Oxidant stress stimulates phosphorylation of eIF4E without an effect on global protein synthesis in smooth muscle cells. Lack of evidence for a role of H202 in angiotensin II-induced hypertrophy. AB - Reactive oxygen species (ROS) are implicated in the pathogenesis of several proliferative diseases, including atherosclerosis and cancer. Eukaryotic translation initiation factor 4E (eIF4E) plays an important role in cell proliferation and differentiation. To gain insight into molecular mechanisms by which ROS influence the pathogenesis of these diseases, I have studied the effect of H(2)O(2), a ROS, on eIF4E phosphorylation. H(2)O(2) induced eIF4E phosphorylation in a dose- and time-dependent manner in growth-arrested smooth muscle cells (SMC). H(2)O(2)-induced eIF4E phosphorylation occurred on serine residues. PD098059, a specific inhibitor of mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) kinase inhibited ERK activities but had no significant effect on eIF4E phosphorylation induced by H(2)O(2). Similarly, SB203580, a specific inhibitor of p38 MAPK, although inhibiting H(2)O(2)-induced p38 MAPK activity, had no effect on H(2)O(2)-induced eIF4E phosphorylation. Calphostin C, a specific inhibitor of protein kinase C, also had no effect on H(2)O(2)-induced eIF4E phosphorylation. In contrast, trifluoperazine, an antagonist of calcium/calmodulin kinases, completely blocked H(2)O(2)-induced eIF4E phosphorylation. In addition, intracellular and extracellular Ca(2+) chelators significantly inhibited H(2)O(2)-induced eIF4E phosphorylation. Despite its ability to induce eIF4E phosphorylation, H(2)O(2) had no significant effect on protein levels and new protein synthesis as compared with control. In contrast, it induced the expression of c-Fos, c-Jun, and HSP70 in a time-dependent manner in SMC. Together, these results suggest that H(2)O(2), a ROS and a cellular oxidant, induces eIF4E phosphorylation in a manner that is dependent on Ca(2+) and Ca(2+)/calmodulin kinases and independent of ERKs, p38 MAPK, and protein kinase C. These results also suggest that enhanced eIF4E phosphorylation by H(2)O(2) appears to be an important event in SMC in response to oxidant stress and that eIF4E phosphorylation may be associated with the translation of a small subset of mRNAs such as c-fos, c-jun, and HSP70 gene mRNAs, whose products may have a critical role in cell survival. PMID- 10828073 TI - Growth hormone (GH)-independent dimerization of GH receptor by a leucine zipper results in constitutive activation. AB - Growth hormone initiates signaling by inducing homodimerization of two GH receptors. Here, we have sought to determine whether constitutively active receptor can be created in the absence of the extracellular domain by substituting it with high affinity leucine zippers to create dimers of the growth hormone receptor (GHR) signaling domain. The entire extracellular domain of the GHR was replaced by the hemagglutinin-tagged zipper sequence of either the c-Fos or c-Jun transcription factor (termed Fos-GHR and Jun-GHR, respectively). Transient transfection of Fos-GHR or Jun-GHR resulted in activation of the serine protease inhibitor 2.1 promoter in Chinese hamster ovary-K1 cells to a level equal to that achieved by fully activated wild type GHR. Furthermore, stable expression of Jun-GHR alone or Fos-GHR and Jun-GHR together in the interleukin 3 dependent BaF-B03 cell line resulted in cell proliferation after interleukin 3 withdrawal at a rate equal to maximally stimulated wild type GHR-expressing cells. Activation of STAT 5b was also observed in Fos-Jun-GHR-expressing cells at a level equal to that in chronically GH-treated GHR-expressing cells. Thus, forced dimerization of the transmembrane and cytoplasmic domains of the GHR in the absence of the extracellular domain can lead to the constitutive activation of known GH signaling end points, supporting the view that proximity of Janus kinase 2 (JAK2) kinases is the essential element in signaling. Such constitutively active GH receptors may have particular utility for transgenic livestock applications. PMID- 10828074 TI - A dominant-negative UBC12 mutant sequesters NEDD8 and inhibits NEDD8 conjugation in vivo. AB - NEDD8, a novel ubiquitin-like protein, has been shown to conjugate to proteins in a manner analogous to ubiquitination and sentrinization. Recently, human UBC12 was identified as a putative NEDD8 conjugation enzyme (E2). While investigating the in vivo function of UBC12, we found that the point mutant, UBC12(C111S), showed a dominant-negative effect on NEDD8 conjugation. This mutant, with a single Cys-to-Ser substitution at the conserved Cys residue in the E2 family, could specifically inhibit NEDD8 conjugation. We observed the dominant-negative effect on NEDD8 conjugation to substrates, including the C-terminal fragment of cullin-2 (Cul-2-DeltaN), full-length cullin-1, and also other uncharacterized target proteins. Interestingly, UBC12(C111S) formed a heterodimeric conjugate with NEDD8. This conjugate was stable under stringent conditions, including 6 m guanidine HCl, 8 m urea, 2% SDS, or 5% beta-mercaptoethanol. Our results are consistent with the hypothesis that UBC12(C111S) sequesters the NEDD8 monomer by forming a UBC12(C111S)-NEDD8 conjugate and, in turn, inhibits the subsequent transfer of NEDD8 to its targets. To examine the biological role of NEDD8 conjugation, this dominant-negative form of UBC12 was applied to a cell growth assay. Overexpression of UBC12(C111S) led to inhibition of growth in U2OS and HEK293 cells. Thus, this dominant-negative form of UBC12 could be useful in defining the role of NEDD8 modification in other biological systems. PMID- 10828075 TI - Transcriptional activation of JC virus by human T-lymphotropic virus type I Tax protein in human neuronal cell lines. AB - Polyomavirus JC (JCV) causes the human demyelinating disease, progressive multifocal leukoencephalopathy (PML). The recent demonstration of cases of PML in association with human T-lymphotropic virus type I (HTLV-I) infection prompted us to examine whether the HTLV-I-encoded regulatory protein Tax activates JCV transcription. By employing a dual luciferase assay, we initially found that the expression of Tax activated the transcriptional potential of both early and late promoters of JCV in human neuronal but not in non-neuronal cells. We subsequently analyzed the mechanism of Tax-induced activation of the JCV promoter in neuronal cells with the following results: 1) the JCV promoter that lacks the NF-kappaB binding motif could not be activated by Tax; 2) the overexpression of IkappaBalpha abolished Tax-induced transcriptional activation of the JCV promoter; 3) a Tax mutant (M22) lacking the potential for activation via the NF kappaB pathway did not activate the JCV promoter. Furthermore, Tax enhances the gene expression of JCV T antigen and VP1. We examined mechanisms of the cell specific activation of the JCV promoter by Tax. Electrophoretic mobility shift assay demonstrated the presence of Tax-bound protein(s) that were specifically present in non-neuronal cells. This study is the first demonstration of the activation of JCV promoter by HTLV-I Tax in an NF-kappaB-dependent manner. PMID- 10828076 TI - Regulation of receptor-mediated protein kinase C membrane trafficking by autophosphorylation. AB - Signal transduction via protein kinase C (PKC) is closely regulated by its subcellular localization. In response to activation of cell-surface receptors, PKC is directed to the plasma membrane by two membrane-targeting domains, namely the C1 and C2 regions. This is followed by the return of the enzyme to the cytoplasm, a process shown recently to require PKC autophosphorylation (Feng, X., and Hannun, Y. A. (1998) J. Biol. Chem. 273, 26870-26874). In the present study, we examined mechanisms of translocation and reverse translocation and the role of autophosphorylation in these processes. By visualizing the trafficking of wild type as well as mutant PKCbetaII in live cells, we demonstrated that in response to cell-surface receptor activation, the function of the C1 region is required but not sufficient for recruitment of the enzyme to the plasma membrane. The C2 region is also critical in anchoring the enzyme to the plasma membrane. Furthermore, the inability of a kinase-deficient PKC to undergo reverse translocation was restored by the addition of intracellular calcium chelators, suggesting a role for the C2 region in the persistent phase of translocation. On the other hand, the inability of a C2 deletion mutant (C1 region intact) to translocate in response to agonist was reversed in mutants lacking kinase activity or by mutation of the Ser(660) autophosphorylation site to alanine, suggesting that autophosphorylation of this site is required for opposing the action of the C2 region. Therefore, the membrane-targeting function of the C1 region is facilitated by the C2 region and appears to be opposed by autophosphorylation. Taken together, these findings provide novel evidence of the functional regulation of reversible PKC membrane localization by autophosphorylation, and they show that the dynamic translocation of PKC in response to agonists is tightly regulated in a collaborative fashion by the C1 and C2 regions in balance with the effects of autophosphorylation. PMID- 10828077 TI - Activation of calpain I converts excitotoxic neuron death into a caspase independent cell death. AB - Glutamate receptor overactivation contributes to neuron death after stroke, trauma, and epileptic seizures. Exposure of cultured rat hippocampal neurons to the selective glutamate receptor agonist N-methyl-d-aspartate (300 microm, 5 min) or to the apoptosis-inducing protein kinase inhibitor staurosporine (300 nm) induced a delayed neuron death. In both cases, neuron death was preceded by the mitochondrial release of the pro-apoptotic factor cytochrome c. Unlike staurosporine, the N-methyl-d-aspartate-induced release of cytochrome c did not lead to significant activation of caspase-3, the main caspase involved in the execution of neuronal apoptosis. In contrast, activation of the Ca(2+)-activated neutral protease calpain I was readily detectable after the exposure to N-methyl d-aspartate. In a neuronal cell-free apoptosis system, calpain I prevented the ability of cytochrome c to activate the caspase cascade by inhibiting the processing of procaspase-3 and -9 into their active subunits. In the hippocampal neuron cultures, the inhibition of calpain activity restored caspase-3-like protease activity after an exposure to N-methyl-d-aspartate. Our data demonstrate the existence of signal transduction pathways that prevent the entry of cells into a caspase-dependent cell death program after the mitochondrial release of cytochrome c. PMID- 10828078 TI - Apolipoprotein A-I: structure-function relationships. AB - The inverse relationship between high density lipoprotein (HDL) plasma levels and coronary heart disease has been attributed to the role that HDL and its major constituent, apolipoprotein A-I (apoA-I), play in reverse cholesterol transport (RCT). The efficiency of RCT depends on the specific ability of apoA-I to promote cellular cholesterol efflux, bind lipids, activate lecithin:cholesterol acyltransferase (LCAT), and form mature HDL that interact with specific receptors and lipid transfer proteins. From the intensive analysis of apoA-I secondary structure has emerged our current understanding of its different classes of amphipathic alpha-helices, which control lipid-binding specificity. The main challenge now is to define apoA-I tertiary structure in its lipid-free and lipid bound forms. Two models are considered for discoidal lipoproteins formed by association of two apoA-I with phospholipids. In the first or picket fence model, each apoA-I wraps around the disc with antiparallel adjacent alpha-helices and with little intermolecular interactions. In the second or belt model, two antiparallel apoA-I are paired by their C-terminal alpha-helices, wrap around the lipoprotein, and are stabilized by multiple intermolecular interactions. While recent evidence supports the belt model, other models, including hybrid models, cannot be excluded. ApoA-I alpha-helices control lipid binding and association with varying levels of lipids. The N-terminal helix 44-65 and the C-terminal helix 210-241 are recognized as important for the initial association with lipids. In the central domain, helix 100-121 and, to a lesser extent, helix 122 143, are also very important for lipid binding and the formation of mature HDL, whereas helices between residues 144 and 186 contribute little. The LCAT activation domain has now been clearly assigned to helix 144-165 with secondary contribution by helix 166-186. The lower lipid binding affinity of the region 144 186 may be important to the activation mechanism allowing displacement of these apoA-I helices by LCAT and presentation of the lipid substrates. No specific sequence has been found that affects diffusional efflux to lipid-bound apoA-I. In contrast, the C-terminal helices, known to be important for lipid binding and maintenance of HDL in circulation, are also involved in the interaction of lipid free apoA-I with macrophages and specific lipid efflux. While much progress has been made, other aspects of apoA-I structure-function relationships still need to be studied, particularly its lipoprotein topology and its interaction with other enzymes, lipid transfer proteins and receptors important for HDL metabolism. PMID- 10828079 TI - Prostaglandin E(2) regulation of cyclooxygenase expression in keratinocytes is mediated via cyclic nucleotide-linked prostaglandin receptors. AB - Inflammatory responses are thought to be mediated in part by the prostaglandins derived from arachidonic acid (AA) by the action of prostaglandin H synthase, also referred to as cyclooxygenase (COX). The mitogen-inducible isoform, COX-2, is over-expressed in numerous chronic inflammatory disease conditions and in neoplasms from both human and experimental animal models. COX-1 expression, on the other hand, has been referred to as constitutive or non-inducible. In this study, we present evidence demonstrating autoregulation of prostaglandin (PG) production by the PGs themselves and their precursor, AA. We observed that AA and PGs induced COX-2, as well as COX-1, expression in cultured murine keratinocytes approximately 3 h after treatment. In primary keratinocytes transiently transfected with a full-length COX-2 promoter linked to a luciferase reporter gene, we observed enhanced transcription by AA, PGE(2), and the other prostaglandins. Forskolin, a known activator of adenylate cyclase, and dibutryl cAMP, a cAMP analog, induced COX-1 and COX-2 mRNA, suggesting that cAMP is a second messenger for COX expression. SQ 22536, an adenylate cyclase inhibitor, inhibited COX-2 mRNA induction by PGE(2) in a dose-dependent manner suggesting that PGE(2)-induced expression may be through one of the cAMP-linked PGE(2) receptors. The results of this study demonstrate that both COX-1 and COX-2 are inducible. Further, both COX isoforms can be up-regulated by their products, the PGs, and this autoregulation probably occurs via prostaglandin receptors linked to a cAMP signal transduction pathway. PMID- 10828080 TI - An immortalized rat liver stellate cell line (HSC-T6): a new cell model for the study of retinoid metabolism in vitro. AB - Hepatocytes and hepatic stellate cells play important roles in retinoid storage and metabolism. Hepatocytes process postprandial retinyl esters and are responsible for secretion of retinol bound to retinol-binding protein (RBP) to maintain plasma retinol levels. Stellate cells are the body's major cellular storage sites for retinoid. We have characterized and utilized an immortalized rat stellate cell line, HSC-T6 cells, to facilitate study of the cellular aspects of hepatic retinoid processing. For comparison, we also carried out parallel studies in Hepa-1 hepatocytes. Like activated primary stellate cells, HSC-T6 express myogenic and neural crest cytoskeletal filaments. HSC-T6 cells take up and esterify retinol in a time- and concentration-dependent manner. Supplementation of HSC-T6 culture medium with free fatty acids (up to 300 micrometer) does not affect retinol uptake but does enhance retinol esterification up to 10-fold. RT-PCR analysis indicates that HSC-T6 cells express all 6 retinoid nuclear receptors (RARalpha, -beta, -gamma, and RXRalpha, -beta, gamma) and like primary stellate cells, HSC-T6 stellate cells express cellular retinol-binding protein, type I (CRBP) but fail to express either retinol-binding protein (RBP) or transthyretin (TTR). Addition of retinol (10(-8)-10(-5) m) or all-trans-retinoic acid (10(-10)-10(-6) m) rapidly up-regulates CRBP expression. Using RAR-specific agonists and antagonists and an RXR-specific agonist, we show that members of the RAR-receptor family modulate HSC-T6 CRBP expression.Thus, HSC T6 cells display the same retinoid-related phenotype as primary stellate cells in culture and will be a useful tool for study of hepatic retinoid storage and metabolism. PMID- 10828081 TI - Cholesterol-sphingomyelin interaction in membrane and apolipoprotein-mediated cellular cholesterol efflux. AB - Helical apolipoproteins interact with cellular surface and generate high density lipoprotein (HDL) by removing phospholipid and cholesterol from cells. We have reported that the HDL is generated by this reaction with the fetal rat astrocytes and meningeal fibroblasts but cholesterol is poorly available to this reaction with the astrocytes (Ito et al. 1999. J. Neurochem. 72: 2362;-2369). Partial digestion of the membrane by extracellular sphingomyelinase increased the incorporation of cholesterol into thus-generated HDL from both types of cell. This increase was diminished by supplement of endogenous or exogenous sphingomyelin to the cells. The sphingomyelinase treatment decreased cholesterol in the membrane mainly in the detergent-resisting domain. The intracellular cholesterol used by acylCoA:cholesterol acyltransferase increased by the sphingomyelinase treatment in the absence of apoA-I, more remarkably in the fibroblast than in the astrocytes. ApoA-I suppressed this increase completely in the astrocytes, but only partially in the fibroblast. The effect of the sphingomyelin digestion was more prominent for the apolipoprotein-mediated reaction than the diffusion-mediated cellular cholesterol efflux. Thus, cholesterol molecules restricted by sphingomyelin in the domain of the plasma membrane appear to be primarily used for the HDL assembly upon the apolipoprotein;-cell interaction. PMID- 10828082 TI - Preparative free-solution isotachophoresis for separation of human plasma lipoproteins: apolipoprotein and lipid composition of HDL subfractions. AB - We have previously shown that plasma lipoproteins can be separated by analytical capillary isotachophoresis (ITP) according to their electrophoretic mobility in a defined buffer system. As in lipoprotein electrophoresis, HDL show the highest mobility followed by VLDL, IDL, and LDL. Chylomicrons migrate according to their net-charge between HDL and VLDL, because ITP has negligible molecular sieve effects. Three HDL subfractions were obtained which were designated fast-, intermediate-, and slow-migrating HDL. To further characterize these HDL subfractions, a newly developed free-solution ITP (FS-ITP)-system was used, that allows micro-preparative separation of human lipoproteins directly from whole plasma (Bottcher, A. et al. 1998. Electrophoresis. 19: 1110-1116). The fractions obtained by FS-ITP were analyzed for their lipid and apolipoprotein composition and by two-dimensional nondenaturing polyacrylamide gradient gel electrophoresis (2D-GGE) with subsequent immunoblotting. fHDL are characterized by the highest proportion of esterified cholesterol of all three subfractions and are relatively enriched in LpA-I. Together with iHDL they contain the majority of plasma apoA-I, while sHDL contain the majority of plasma apoA-IV, apoD, apoE, and apoJ. Pre-beta HDL were found in separate fractions together with triglyceride-rich fractions between sHDL and LDL. In summary, ITP can separate the bulk of HDL into lipoprotein subfractions, which differ in apolipoprotein composition and electrophoretic mobility. While analytical ITP permits rapid separation and quantitation for diagnostic purposes, FS-ITP can be used to obtain these lipoprotein subfractions on a preparative scale for functional analysis. As FS ITP is much better suited for preparative purposes than gel electrophoresis, it represents an important novel tool for the functional analysis of lipoprotein subclasses. PMID- 10828083 TI - Sphingomyelin exhibits greatly enhanced protection compared with egg yolk phosphatidylcholine against detergent bile salts. AB - Inclusion of phosphatidylcholine within bile salt micelles protects against bile salt-induced cytotoxicity. In addition to phosphatidylcholine, bile may contain significant amounts of sphingomyelin, particularly under cholestatic conditions. We compared protective effects of egg yolk phosphatidylcholine (similar to phosphatidylcholine in bile), egg yolk sphingomyelin (mainly 16:0 acyl chains) and dipalmitoyl phosphatidylcholine against taurocholate in complementary in vitro studies. Upon addition of taurocholate-containing micelles to sonicated egg yolk phosphatidylcholine vesicles, subsequent micellization of the vesicular bilayer proved to be retarded when phospholipids had also been included in these micelles in the rank order: egg yolk phosphatidylcholine < dipalmitoyl phosphatidylcholine < sphingomyelin. Hemolysis of erythrocytes and LDH release by CaCo-2 cells after addition of taurocholate micelles were strongly reduced by including small amounts of sphingomyelin or dipalmitoyl phosphatidylcholine in these micelles (PL/(PL + BS) >/= 0.1), whereas egg yolk phosphatidylcholine provided less protection. Amounts of non-phospholipid-associated bile salts (thought to be responsible for cytotoxicity) in egg yolk phosphatidylcholine containing micelles were significantly higher than in corresponding sphingomyelin or dipalmitoyl phosphatidylcholine-containing micelles (tested at PL/(PL + BS) ratios 0.1, 0.15, and 0.2). LDH release upon incubation of CaCo-2 cells with taurocholate simple micelles at these so-called "intermixed micellar-vesicular" concentrations was identical to LDH release upon incubation with corresponding taurocholate-phospholipid mixed micelles. In conclusion, we found greatly enhanced protective effects of sphingomyelin and dipalmitoyl phosphatidylcholine compared to egg yolk phosphatidylcholine against bile salt-induced cytotoxicity, related to different amounts of non-phospholipid-associated bile salts. These findings may be relevant for protection against bile salt-induced cytotoxicity in vivo. PMID- 10828084 TI - Effect of growth hormone replacement therapy on plasma lecithin:cholesterol acyltransferase and lipid transfer protein activities in growth hormone-deficient adults. AB - The effects of growth hormone (GH) replacement on plasma lecithin:cholesterol acyltransferase (LCAT), cholesteryl ester transfer protein (CETP), and phospholipid transfer protein (PLTP), factors involved in high density lipoprotein (HDL) metabolism, are unknown. We carried out a 6 months study in 24 GH-deficient adults who were randomized to placebo (n = 8), low dose GH (1 U daily, n = 8), and high dose GH (2 U daily, n = 8), followed by a 6 months open extension study with high dose GH (1 drop-out). No significant changes in plasma lipoproteins, LCAT, CETP, and PLTP activities, cholesterol esterification (EST) and cholesteryl ester transfer (CET) were observed after placebo. After 6 months of GH (combined data, n = 24), very low + low density lipoprotein (VLDL + LDL) cholesterol (P < 0.05) and apolipoprotein B (P < 0.05) decreased, whereas HDL cholesterol and HDL cholesteryl ester increased (P < 0. 05). Prolonged treatment showed comparable effects. Plasma apolipoprotein A-I and Lp[a] remained unchanged. Plasma LCAT (P < 0. 01) and CETP activities (P < 0.01), as well as EST (P < 0.01) and CET decreased (P < 0.01) after 12 months of GH (n = 15), but PLTP activity did not significantly change. Changes in EST and CET after 12 months of treatment were independently related to changes in plasma LCAT (P = 0.001 and CETP activity (P = 0.01). In conclusion, GH replacement therapy improves the lipoprotein profile in GH-deficient adults. Chronic GH replacement lowers plasma LCAT and CETP activities, contributing to a decrease in cholesterol esterification and cholesteryl ester transfer. These effects may have consequences for HDL metabolism and reverse cholesterol transport. PMID- 10828085 TI - alpha-cyclodextrin extracts diacylglycerol from insect high density lipoproteins. AB - alpha-Cyclodextrins are water-soluble cyclic hexamers of glucose units with hydrophobic cavities capable of solubilizing lipophiles. Incubating alpha cyclodextrin with high density lipophorin from Manduca sexta or Bombyx mori resulted in a cloudy, turbid solution. Centrifugation separated a pale yellowish precipitate. Thin-layer chromatography analysis of the lipid extract of the precipitate showed that the major lipid was diacylglycerol, while KBr density gradient analysis of the supernatant demonstrated the presence of a lipid depleted very high density lipophorin. Transfer of diacylglycerol from lipophorin to cyclodextrin was specific to alpha-cyclodextrin and was not observed with beta or gamma-cyclodextrins. pH had no effect on diacylglycerol transfer to alpha cyclodextrin. However, the transfer was strongly dependent on the concentration of alpha-cyclodextrin and temperature. Increasing the concentration of alpha cyclodextrin in the incubation mixture was associated with the formation of increasingly higher density lipophorins. Thus, at 20, 30, and 40 mm alpha cyclodextrin, the density of B. mori lipophorin increased from 1.107 g/ml to 1.123, 1. 148, and 1.181 g/ml, respectively. At concentrations greater than 40 mm, alpha-cyclodextrin had no further effect on the density of lipophorin. alpha Cyclodextrin removed at most 83;-87% of the diacylglycerol present in lipophorin. Temperature played an important role in altering the amount of diacylglycerols transferred to alpha-cyclodextrin. At 30 mm alpha-cyclodextrin, the amount of diacylglycerol transferred at different temperatures was 50% at 4 degrees C, 41% at 15 degrees C, 20% at 28 degrees C, and less than 3% at 37 degrees C. We propose that diacylglycerol transfers to alpha-cyclodextrin via an aqueous diffusion pathway and that the driving force for the transfer is the formation of an insoluble alpha-cyclodextrin-diacylglycerol complex. PMID- 10828086 TI - Specific inhibition of rat brain phospholipase D by lysophospholipids. AB - Although the importance of phospholipase D (PLD) in signal transduction in mammalian cells is well documented, the negative regulation of PLD is poorly understood. This is primarily due to a lack of known specific inhibitors of PLD. We herein report that the activity of partially purified rat brain PLD is inhibited by certain lysophospholipids, such as lysophosphatidylinositol, lysophosphatidylglycerol, and lysophosphatidylserine in a highly specific manner. Inhibition of PLD by lysophospholipids was dose-dependent: the concentration of lysophosphatidylinositol required for half-maximal inhibition was about 3 micrometer. An analysis of the enzyme-kinetics suggested that lysophospholipids act as non-competitive inhibitors of PLD activity. As expected, PLD activity was stimulated by ADP-ribosylation factor (Arf) and phosphatidylinositol 4,5 bisphosphate (PIP(2)). The inhibition of PLD by lysophospholipids, however, was not affected by the presence or absence of Arf or by an increase in PIP(2) concentration. A protein-binding assay suggested that lysophospholipids bind directly to PLD. These results indicate that the observed inhibition of PLD by lysophospholipids is due to their direct interaction rather than to an interaction between lysophospholipids and either Arf or PIP(2). The present study suggests that certain lysophospholipids are specific inhibitors of rat brain PLD in a cell-free system and may provide the new opportunities to investigate mechanisms by which PLD is regulated by lysophospholipids, presumably liberated by phospholipase A(2) activation, in mammalian cells. PMID- 10828087 TI - Molecular scanning of the human PPARa gene: association of the L162v mutation with hyperapobetalipoproteinemia. AB - Peroxisome proliferator-activated receptor alpha (PPARalpha) is a member of the steroid hormone receptor super family involved in the control of cellular lipid utilization. This makes PPARalpha a candidate gene for type 2 diabetes and dyslipidemia. The aim of this study was to investigate whether genetic variation in the human PPARalpha gene can influence the risk of type 2 diabetes and dyslipidemia among French Canadians. We therefore first determined the genomic structure of human PPARalpha, and then designed intronic primers to sequence the coding region and the exon-intron boundaries of the gene in 12 patients with type 2 diabetes and in 2 nondiabetic subjects. Sequence analysis revealed the presence of a L162V missense mutation in exon 5 of one diabetic patient. Leucine 162 is contained within the DNA binding domain of the human PPARalpha gene, and is conserved among humans, mice, rats, and guinea pigs. We subsequently screened a sample of 121 patients newly diagnosed with type 2 diabetes and their age and sex matched nondiabetic controls, recruited from the Saguenay-Lac-St-Jean region of Northeastern Quebec, for the presence of the L162V mutation by a PCR-RFLP based method. There was no difference in L162 homozygote or V162 carrier frequencies between diabetics and nondiabetics. However, whether diabetic or not, carriers of the V162 allele had higher plasma apolipoprotein B levels compared to noncarriers (P 5 0.05). To further this association, we screened another sample of 193 nondiabetic subjects recruited in the greater Quebec City area. Carriers of the V162 allele compared with homozygotes of the L162 allele had significantly higher concentrations of plasma total and LDL-apolipoprotein B as well as LDL cholesterol (P 0.29%, 8 hr) and penetration (2.48-->1.11%). In conclusion, dermal absorption data from liquid-organic or aqueousorganic mixtures may underestimate the risk of exposure to TCB-contaminated soil. PMID- 10828143 TI - Effects of industrial detergents on the barrier function of human skin. AB - Detergents are involved in the causation of contact dermatitis and in promoting percutaneous absorption of toxic chemicals, but limited information is available to allow an assessment of their relative effects on the skin barrier function. The effect of detergents on skin permeability to water and nickel was examined in an in-vitro model using human skin. Twenty-four of the most widely used detergents were studied. After a two-hour exposure to an aqueous detergent solution, penetration of labeled model compounds was followed for 66 hours. Interindividual variation was substantial, but 12 of the detergents caused statistically significant increases in the penetration of water, nickel, or both. Nonionic detergents were as likely as anionic detergents to have this effect. This study demonstrates that useful information may be obtained by a simple in vitro method, and that such data may provide a basis for substitution efforts. PMID- 10828144 TI - Effects of four detergents on the in-vitro barrier function of human skin. AB - The key to promoting percutaneous penetration is to alter the properties of the dermal barrier function. Mechanisms for changing the barrier function vary. Thus, some enhancers induce reversible conformational changes, whereas others cause prolonged barrier disruption. Discrimination between the influences of detergents on the barrier function may enable the use of those detergents affecting the overall integrity of the skin the least. In an experimental setup using in-vitro static diffusion cells mounted with human skin, the effects of four extensively used detergents on dermal barrier function were evaluated. Three of the detergents (lutensol AP10, nonyl phenyl ethoxylate, ethanol) apparently enhance percutaneous penetration without compromising the overall integrity of the skin barrier. SLS, on the other hand, acts through a time-and-dose-dependent deterioration of the dermal barrier function. The present experimental approach allows discrimination between detergents that act through different mechanisms to enhance percutaneous penetration. PMID- 10828145 TI - Exposure and absorption of hazardous materials through the skin. PMID- 10828146 TI - Manganese--a public health concern: its relevance for occupational health and safety policy and regulation in South Africa. AB - Concerns about the effects of low-level manganese exposures on human health arise at a time when South Africa finds itself in competition with newcomers to the market economy, China and the CIS. This case study illustrates how decisions about occupational health and safety and the environment are influenced by incompleteness of scientific knowledge, competing interests, differences over what is fair or just, and the compartmentalization of public policy. In addition, an assessment is made of the ability of the occupational health and safety system in South Africa in its current form to address the challenges posed by manganese related issues. The importance of tracking developments abroad, strengthening participatory processes, developing national policy, linking economic policy and OHS policy, and establishing appropriate trade agreements is stressed. PMID- 10828147 TI - Seattle and the ICOH: the view from the other side of the globe. PMID- 10828148 TI - The international ban asbestos secretariat. PMID- 10828149 TI - An adaptive system for active noise reduction. AB - An adaptive system for active noise reduction in an acoustic duct is presented. The system is based on a modification of a least mean square (LMS) algorithm called filtered-U with on-line error path modelling. The system was assembled and examined on a laboratory test stand in the Laboratory of Active Noise Reduction Methods of the Central Institute for Labour Protection (Warsaw, Poland). The structure of the test stand, the block structure of the active noise reduction system, the basic assumption concerning the applied adaptive algorithm, and examples of measured effectiveness of the system for various kinds of noise are presented. PMID- 10828150 TI - Measurement of the response time of an electrosensitive protective device in the process of its certification. AB - In the process of testing an electrosensitive protective system, determining its response time is of crucial importance. A unique double penetration method of measuring electrosensitive protective device (ESPD) response time has been worked out in the Central Institute for Labour Protection. In the first step, low speed penetration enables the detection zone border to be localised. In the second step of measurement, the probe is injected at a high speed and response time is measured. Three different ways have been taken for validation of the method: theoretical analysis, calibration of the stand, and taking a series of measurements. The double penetration method enables ESPD response time measurement results to be obtained, the accuracy, repeatability, and reproducibility of which is satisfactory enough to be assessed objectively. PMID- 10828151 TI - Prediction of toxic substances emission for occupational exposure assessment. AB - Methods for predicting organic solvents, chromic acid, mineral oil, styrene, and sulphuric acid emissions in painting, metal degreasing, wood preservation, chromium electroplating, turning, grinding, making glass fortified polyester laminates and lead batteries charging, injection moulding of polystyrene plastics, and making polyurethane foam processes are described. Experimentally introduced equations are based on the essential parameters of these processes. Knowing the emission and the total flow rate of ventilation, it is possible to calculate toxic agent concentration, which is the basis of occupational exposure assessment. PMID- 10828152 TI - A study of the nonsteady-state filtration process in a fibrous material in conditions of real dust loading. AB - The study concerns ways to describe filtration efficiency and gas flow resistance while particles are deposited in fibrous respiratory protective filters under conditions characteristic for their use. The performance of fibrous filter materials of varied structure and electrostatic properties was studied, using polydisperse coarse-grained aerosols and submicron aerosols of sodium chloride. Evidence was found that changes in airflow resistance depended to a large extent on air humidity and the concentration of the aerosol. A relationship was also found between the electrostatic properties of filter media and their decreasing efficiency with time. A numerical model was developed to simulate the phenomenon of nonsteady-state filtration for 2 mechanisms of deposition, which permitted assessment of the filtration characteristics of filter media of any design and in any conditions. PMID- 10828154 TI - Impact of economic incentives on costs and benefits of occupational health and safety. AB - The most common type of economic incentive used in the field of health and safety is experience rating of insurance premiums. The impact of this incentive on occupational health and safety (OHS) costs in the company was analysed by comparing insurance costs with other OHS costs associated with inadequate working conditions, such as accident costs borne by a company. Accident costs were estimated on the basis of research carried out in 10 companies. Insurance costs and their adjustments according to the health and safety level in a company were calculated according to an experience rating model developed in the Central Institute for Labour Protection. PMID- 10828153 TI - Monitoring psychosocial stress at work: development of the Psychosocial Working Conditions Questionnaire. AB - Many studies on the impact of psychosocial working conditions on health prove that psychosocial stress at work is an important risk factor endangering workers' health. Thus it should be constantly monitored like other work hazards. The paper presents a newly developed instrument for stress monitoring called the Psychosocial Working Conditions Questionnaire (PWC). Its structure is based on Robert Karasek's model of job stress (Karasek, 1979; Karasek & Theorell, 1990). It consists of 3 main scales Job Demands, Job Control, Social Support and 2 additional scales adapted from the Occupational Stress Questionnaire (Elo, Leppanen, Lindstrom, & Ropponen, 1992), Well-Being and Desired Changes. The study of 8 occupational groups (bank and insurance specialists, middle medical personnel, construction workers, shop assistants, government and self-government administration officers, computer scientists, public transport drivers, teachers, N = 3,669) indicates that PWC has satisfactory psychometrics parameters. Norms for the 8 groups were developed. PMID- 10828155 TI - Occupational health and safety management in polish enterprises implementing total quality management systems. AB - Total Quality Management (TQM) is defined as the management approach of the organization aimed at long-term success through client satisfaction, and which benefits all members of the organization and society (ISO 8402; International Organization for Standardization, 1994a). The objective of the study was to evaluate management methods applied to improve working conditions in Polish enterprises implementing TQM. The investigation was conducted in the form of interviews, which covered relevant connections between the TQM concept and occupational health and safety (OHS) systematic management rules. The results revealed that the criteria adopted in investigated enterprises for OHS management systems, as well as the implemented management methods and tools, can be evaluated positively. However, many require significant improvement in order to ensure better compliance with the existing law provisions. Elements of OHS management systems also require better integration with the overall management system of the enterprise. PMID- 10828156 TI - Polish system of assessing occupational risk posed by chemical compounds. AB - According to the Polish Labour Code &elpar;Ustawa, 1974) employers are legally obligated to provide workers with information about occupational health and safety risks. Maximum allowable concentrations (MAC) and the results of determining chemical compounds in workplace air are used for assessing occupational exposure and risk. A computer-assisted system STER, developed in the Central Institute for Labour Protection, helps to register and document occupational risk assessment and all actions resulting from those assessments. PMID- 10828157 TI - Preferred levels of auditory danger signals. AB - An important issue at the design stage of the auditory danger signal for a safety system is the signal audibility under various conditions of background noise. The auditory danger signal should be clearly audible but it should not be too loud to avoid fright, startling effects, and nuisance complaints. Criteria for designing auditory danger signals are the subject of the ISO 7731 (International Organization for Standardization [ISO], 1986) international standard and the EN 457 European standard (European Committee for Standardization [CEN], 1992). It is required that the A-weighted sound pressure level of the auditory danger signal is higher in level than the background noise by 15 dB. In this paper, the results of an experiment are reported, in which listeners adjusted most preferred levels of 3 danger signals (tone, sweep, complex sound) in the presence of a noise background (pink noise and industrial noise). The measurements were done for 60-, 70-, 80-, and 90-dB A-weighted levels of noise. Results show that for 60-dB level of noise the most preferred level of the danger signal is 10 to 20 dB above the noise level. However, for 90-dB level of noise, listeners selected a level of the danger signal that was equal to the noise level. Results imply that the criterion in the existing standards is conservative as it requires the level of the danger signal to be higher than the level of noise regardless of the noise level. PMID- 10828158 TI - Practical application of ergonomic settings of typical computerised workstations. AB - The goal of the study was to check, with regard to ergonomics, workstations equipped with visual display terminals in selected enterprises. Over 180 workstations were tested in 3 enterprises. Most workstations were equipped with computers. The ergonomics of both the parameters of the basic components of the workstation (i.e., a chair and a desk, and the position of the computer at the workstation and its screen with respect to windows) and lighting fittings were analysed. Typical mistakes in the layout of a workstation were chairs inappropriate for computer work, as well as broken chair adjustment mechanisms, which qualified chairs for repair or replacement. Wrong positioning of monitors on the desk and with regard to windows and lighting fittings was also noted. PMID- 10828159 TI - Assessment of psychophysical abilities of people with disabilities during occupational rehabilitation. AB - Designing objective and subjective methods of studying the abilities of disabled people is one of the most important things to be done before employing them. That is why, new assessment methods are proposed. Evaluation as a testing procedure used for defining the abilities of disabled people general capacity, range, and strength of the main groups of muscles and extremities, psychological tests, and many others were developed by an interdisciplinary team. In total, 48 testing procedures, including physiological, psychological, and biomechanical ones, were prepared. We also propose another subjective assessment method because it is also very important to know what an individual person with disability needs, wants to do, or what kind of job he or she prefers. The subjective evaluation of abilities, possibilities, and needs might be done by using a questionnaire. These testing procedures offer a chance to use the same methods in all diagnostic centres so the criteria of health and work ability will be the same and easily comparable. Using the same tests for assessment will be helpful in observing the results and progress of medical, social, and vocational rehabilitation, too. PMID- 10828160 TI - Thirst and work capacity of older people in a hot environment. AB - In a hot environment, especially during exercise, the main role of thermoregulative mechanisms is to dissipate excessive heat from the body. The most effective way of heat dissipation is sweat production and its evaporation off skin surface. Intense sweating results in a considerable loss of water and electrolytes. There are some results that indicate lower thirst of older men than young ones in response to a hot environment and osmotic stimuli. Our studies conducted in men of different ages exposed to a hot environment indicated that there were no significant differences in rectal temperature (Tre) or heat storage (S) among groups at rest. Lower Tre and lower S in older men with higher physical capacity for their age than in young ones with average physical capacity was shown. However, in all the experiments significantly lower thirst in older men than in the young was indicated. Special attention should be paid to fluid replacement among workers in a hot environment because of lower thirst and lower hydration of older men. These individuals may be exposed to dehydration risk during prolonged exercise, despite the possibility to tolerate heat strain as well as young ones. Further studies are necessary because of some diversity in the meaning of the results. PMID- 10828161 TI - Review of research studies of ergonomic aspects of selected personal protective equipment. AB - This article is a review of research studies conducted in the Central Institute for Labour Protection (CIOP) on ergonomic aspects of personal protective equipment (PPE) design and use. Research was related to, among other things, a comparison of body strain and work time limitation for users wearing gas-tight and drill suits. Some other studies were connected with the biophysical properties of clothing materials for medical use. On the basis of the measurement results, a proposal of clothing construction with an optimum combination of protection and comfort was created. Research leading to the development of test methodology concerning automatic welding filters with switchable luminous transmittance is also conducted in CIOP. The article also brings up the subject of designing new PPE assuring a high level of comfort. A model of material to be used under tight protective clothing, an inflammable harness for working at a height, and a model of intelligent clothing that will assist human thermoregulation processes during physical effort are examples of new PPE. PMID- 10828162 TI - A Scientific Meeting held at the Royal College of Physicians, London on 27 January, 1999. AB - These Abstracts of Original Communications are presented by the authors as camera ready artwork ready for printing, and are therefore only available in PDF format. The HTML "full text availability" lists the titles and authors of each paper, and the PDF file presents the text. PMID- 10828163 TI - A Scientific Meeting held at Maastricht University, Maastricht, The Netherlands on 18-21 February 1999. AB - These Abstracts of Original Communications are presented by the authors as camera ready artwork ready for printing, and are therefore only available in PDF format. The HTML "full text availability" lists the titles and authors of each paper, and the PDF file presents the text. PMID- 10828165 TI - An appreciation. PMID- 10828166 TI - An appreciation. PMID- 10828164 TI - Obituary. PMID- 10828168 TI - Proceedings of the nutrition society: 2000 and beyond PMID- 10828167 TI - The 6th International Tutorial Conference of the Research Group on the Biochemistry of Exercise, a collaboration between the Nutrition Toxicology and Environmental Research Institute Maastricht and the Nutrition Society was held in Maastricht, The Netherlands on 18-21 February 1999. PMID- 10828169 TI - Essential fatty acids in early life: structural and functional role. AB - Essential fatty acids (EFA) are structural components of all tissues and are indispensable for cell membrane synthesis; the brain, retina and other neural tissues are particularly rich in long-chain polyunsaturated fatty acids (LCPUFA). These fatty acids serve as specific precursors for eicosanoids that regulate numerous cell and organ functions. Results from animal and recent human studies support the essential nature of n-3 EFA in addition to the well-established role of n-6 EFA for human subjects, particularly in early life. The most significant effects relate to neural development and maturation of sensory systems. Recent studies using stable-isotope-labelled tracers demonstrate that even preterm infants are able to form arachidonic acid (AA) and docosahexaenoic acid (DHA), but that synthesis is extremely low. Intracellular fatty acids or their metabolites regulate transcriptional activation of gene expression during adipocyte differentiation, and retinal and nervous system development. Regulation of gene expression by LCPUFA occurs at the transcriptional level and is mediated by nuclear transcription factors activated by fatty acids. These nuclear receptors are part of the steroid hormone receptor family. Two types of polyunsaturated fatty acid responsive transcription factors have been characterized, the peroxisome proliferator-activated receptor (PPAR) and the hepatic nuclear factor 4alpha. DHA also has significant effects on photoreceptor membranes involved in the signal transduction process, rhodopsin activation, and rod and cone development. Comprehensive clinical studies have shown that dietary supplementation with marine oil or single-cell oils, sources of LCPUFA, results in increased blood levels of DHA and AA, as well as an associated improvement in visual function in formula-fed premature infants to match that of human milk-fed infant. Recent clinical trials convincingly support LCPUFA supplementation of preterm infant formulations and possibly term formula to mimic human milk composition. PMID- 10828170 TI - Long-term adequacy of exclusive breast-feeding: how scientific research has led to revised opinions. AB - The present paper reviews the research of the authors and their colleagues over the past 20 years to provide improved nutritional and anthropometric guidelines for the assessment of lactational adequacy and for when the weaning process might be initiated. The nutritional guidelines are based on revised dietary energy requirements. The basic assumption is that since breast milk is a well-balanced food, if energy needs are satisfied so will those for essential nutrients. Energy requirements for young babies have been derived from the application of the doubly-labelled-water technique. This approach indicated that 460 kJ (110 kcal)/kg per d at 1 month and 397 and 355 kJ (95 and 85 kcal)/kg per d at 3 and 6 months respectively would be satisfactory for a nutrient content of high bioavailability. Translated into a breast-milk intake of 850 ml/d the latter would cover the dietary energy needs of the average child growing along the 50th centile until at least 4 months, but the typical child from many developing countries following the 25th centile until 6 months. The importance of revised growth reference values for infancy, equally crucial for assessing lactational adequacy, is also reviewed. In contrast with the shapes of earlier reference patterns, growth trajectories are different when babies are fed in accordance with modern paediatric advice. Mothers and health professionals using the older growth charts to assess the progress of a baby can be misled into assuming that the weaning process needs to be introduced sooner than necessary. Examples of this situation within the context of a developing country are provided. PMID- 10828171 TI - The role of care in nutrition programmes: current research and a research agenda. AB - The importance of cultural and behavioural factors in children's nutrition, particularly with regard to feeding, has been recognized only recently. The combination of evidence regarding the importance of caregiving behaviour for good nutrition, and improved strategies for measuring behaviour have led to a renewed interest in care. The UNICEF conceptual framework suggests that care, in addition to food security and health care services, are critical for children's survival, growth and development. The present paper focuses on the care practice of complementary feeding, specifically behavioural factors such as parental interaction patterns, feeding style and adaptation of feeding to the child's motor abilities (self-feeding or feeding by others). Three kinds of feeding styles (Birch & Fisher, 1995) are identified: controlling; laissez-faire; responsive. Probable effects of each feeding style on nutrient intake are described. A number of studies of feeding behaviour have suggested that the laissez-faire style is most frequently observed among families and communities with a higher prevalence of malnourished children. Nutrition interventions that have been able to show significant effects on outcomes, such as the Hearth Model in Vietnam (Sternin et al. 1997), have usually incorporated behavioural components in their intervention. At this time, there have been no tests of the efficacy of behavioural interventions to improve feeding practices. Research is needed to understand behavioural factors in complementary feeding, and to identify and test intervention strategies designed to improve nutrient intake of young children. Finally, the paper concludes with a discussion of how nutrition programmes might change if care were incorporated. PMID- 10828172 TI - Failure to thrive in a population context: two contrasting studies of feeding and nutritional status. AB - Although failure to thrive (FTT) is generally thought to be a nutritional problem, dietary intake in children with FTT has been little researched. We describe two community-based studies of dietary intake and eating behaviour in FTT. The first study of ninety-seven children with FTT identified by population screening found that only a minority of case children were associated with neglect, organic illness or deprivation, even though dietary information suggested an underlying nutritional cause in the majority. Limited case-control data suggested significantly delayed weaning and less liking for food in general among the case children. The second study found diminished appetite, delayed progression onto solid foods and fewer foods eaten by forty-four children referred with FTT compared with forty-five controls, but was unable to detect a significant difference in energy intake, even allowing for varying body composition and other confounding variables. This study also revealed the bias introduced by recruiting case children by referral rather than screening, and that despite rigorous matching procedures the controls were not representative of the general population. These studies suggest a wide range of differences in eating behaviour and feeding patterns between children with FTT and controls, but suggest that measurements of nutrient totals may not be robust. Studies of FTT based on referred children can be misleading, as can comparison with controls, unless allowance can be made for important confounding social variables. PMID- 10828173 TI - Nutritional deficiencies and later behavioural development. AB - The literature on the long-term effects of nutritional deficiencies in early life is reviewed. The severity and duration of the deficiency, the stage of the children's development, the biological condition of the children and the socio cultural context may all modify the effect. There is substantial evidence that reduced breast-feeding, small-for-gestational-age birth weight, Fe and I deficiency, and protein-energy malnutrition (PEM) are associated with long-term deficits in cognition and school achievement. However, all these conditions are associated with poverty and poor health, which may account for the association. It is difficult to establish that the long-term relationship is causal, as it requires a randomized treatment trial with long-term follow-up. Such studies are only available for I deficiency in utero and early childhood PEM. Results from these studies indicate that I deficiency has a long-term effect and PEM probably has a long-term effect. PMID- 10828174 TI - Factors affecting newborn bone mineral content: in utero effects on newborn bone mineralization. AB - Several factors have been found recently to have a significant impact on newborn bone mineral content (BMC) and developing fetal bone. Recently we showed that maternal vitamin D deficiency may affect fetal bone mineralization. Korean winter born newborn infants had extremely low serum 25-hydroxyvitamin D (25-OHD), high serum cross-linked carboxy-terminal telopeptide of type I collagen (ICTP; a bone resorption marker), and markedly lower (8 %) total body BMC than summer-born newborn infants. Infant total body BMC was positively correlated with cord serum 25-OHD and inversely correlated with ICTP, which was also negatively correlated with vitamin D status. In three separate studies on North American neonates we found markedly lower (8-12 %) BMC in summer newborn infants compared with winter newborn infants, the opposite of the findings for Korean neonates. The major reason for the conflicting BMC results might be the markedly different maternal vitamin D status of the North American and Korean subjects. Recently, we found evidence of decreased bone formation rates in infants who were small-for gestational age (SGA) compared with infants who were appropriate-for-gestational age; we reported reduced BMC, cord serum osteocalcin (a marker of bone formation) and 1,25-dihydroxyvitamin D (the active metabolite of vitamin D), but no alterations in indices of fetal bone collagen metabolism. In theory, reduced utero-placental blood flow in SGA infants may result in reduced transplacental mineral supply and reduced fetal bone formation. Infants of diabetic mothers (IDM) have low BMC at birth, and infant BMC correlated inversely with poor control of diabetes in the mother, specifically first trimester maternal mean capillary blood glucose concentration, implying that factors early in pregnancy might have an effect on fetal BMC. The low BMC in IDM may be related to the decreased transplacental mineral transfer. Cord serum ICTP concentrations were higher in IDM than in control subjects, implying increased intrauterine bone resorption. BMC is consistently increased with increasing body weight and length in infants. Race and gender differences in BMC appear in early life, but not at birth. Ethanol consumption and smoking by the mother during pregnancy affect fetal skeletal development. PMID- 10828175 TI - The role of retinoic acid in embryonic and post-embryonic development. AB - Retinoic acid (RA) is the bioactive metabolite of vitamin A (retinol) which acts on cells to establish or change the pattern of gene activity. Retinol is converted to RA by the action of two types of enzyme, retinol dehydrogenases and retinal dehydrogenases. In the nucleus RA acts as a ligand to activate two families of transcription factors, the RA receptors (RAR) and the retinoid X receptors (RXR) which heterodimerize and bind to the upstream sequences of RA responsive genes. Thus, in addition to the well-established experimental paradigm of depriving animals of vitamin A to determine the role of RA in embryonic and post-embryonic development, molecular biology has provided us with two additional methodologies: knockout the enzymes or the RAR and RXR in the mouse embryo. The distribution of the enzymes and receptors, and recent experiments to determine the endogenous distribution of RA in the embryo are described here, as well as the effects on the embryo of knocking out the enzymes and receptors. In addition, recent studies using the classical vitamin A-deprivation technique are described, as they have provided novel insights into the regions of the embryo which crucially require RA, and the gene pathways involved in their development. Finally, the post-embryonic or regenerating systems in which RA plays a part are described, i.e. the regenerating limb, lung regeneration, hair cell regeneration in the ear and spinal cord regeneration in the adult. PMID- 10828176 TI - The role of iodine in brain development. AB - I is required for the synthesis of thyroid hormones. These hormones, in turn, are required for brain development, which occurs during fetal and early postnatal life. The present paper reviews the impact of I deficiency (1) on thyroid function during pregnancy and in the neonate, and (2) on the intellectual development of infants and children. All extents of I deficiency (based on I intake (microgram/d); mild 50-99, moderate 20-49, severe > 20) affect the thyroid function of the mother and neonate, and the mental development of the child. The damage increases with the extent of the deficiency, with overt endemic cretinism as the severest consequence. This syndrome combines irreversible mental retardation, neurological damage and thyroid failure. Maternal hypothyroxinaemia during early pregnancy is a key factor in the development of the neurological damage in the cretin. Se deficiency superimposed on I deficiency partly prevents the neurological damage, but precipitates severe hypothyroidism in cretins. I deficiency results in a global loss of 10-15 intellectual quotient points at a population level, and constitutes the world's greatest single cause of preventable brain damage and mental retardation. PMID- 10828177 TI - Are milk polyamines preventive agents against food allergy? AB - Insufficient polyamine intake could play a role in the induction of sensitization to dietary allergens. This proposal is based essentially on investigations made in sucking rats and in children. In sucking rats it has been established that oral administration of spermine can induce all the modifications occurring in the digestive tract at weaning. In the intestine events occur in two phases. The early event consists of desquamation of the epithelium resulting from an activation of apoptosis. The late event appears to involve an hormonal cascade in which adrenocorticotropic hormone, cytokines, bombesin and corticosterone are included. Observations in human subjects show that: (1) the spermine and spermidine concentrations are generally lower in infant formulas than in human breast milk. Mothers seem consistently to have relatively high or relatively low concentrations of spermine and spermidine in their milk. These individual variations may be due to diet, lifestyle or genetic background; (2) the probability of developing allergy can reach 80 % if the mean spermine concentration in the milk is lower than 2 nmol/ml milk. It is approximately 0 % if the mean spermine concentration is higher than 13 nmol/ml milk; (3) preliminary results show that the intestinal permeability to macromolecules differs in premature babies when they are fed on breast milk compared with infant formulas (J Senterre, J Rigo, G Forget, G Dandrifosse and N Romain, unpublished results). This difference does not seem to be present when powdered milk is supplemented with polyamines at the concentration found in breast milk; (4) spermine increases proliferation and differentiation of lymphocytes isolated from the tonsils of children. PMID- 10828178 TI - Protein nutrition of the neonate. AB - The period of growth and development between birth and weaning is crucial for the long-term well-being of the organism. Protein deposition is very rapid, is achieved with a high nutritional efficiency, and is accompanied by marked differences in the growth rates of individual tissues and a series of maturational processes. These important aspects of development occur while the neonate is consuming a single and highly-specific food source, milk. Surprisingly, although there is a clear relationship between the nutrient density of milk and the growth rate of its recipient, this relationship does not apply to the overall amino acid composition of mixed milk proteins. Some amino acids, notably glycine and arginine, are supplied in milk in quantities that are much less than the needs of the neonate. The milk-fed neonate is therefore capable of carrying out a tightly-regulated transfer of N from amino acids in excess to those that are deficient. The rapid growth of the neonate is supported by a high rate of tissue protein synthesis. This process appears to be activated by the consumption of the first meals of colostrum. Recent research has identified that skeletal muscle and the brain are specifically responsive to an unidentified factor in colostrum. Following the initial anabolic response the rate of protein synthesis in some tissues, notably muscle, falls from birth to weaning. This decrease reflects a progressively smaller anabolic response to nutrient intake, which not only involves an overall fall in the capacity for protein synthesis, but also in responses to insulin and amino acids. The study of growth and protein metabolism, and their regulation in the neonate is not only important for pediatrics, but may provide important pointers to more general aspects of regulation that could be applied to the nutrition of the mature animal. PMID- 10828179 TI - Use of maternal reserves as a lactation strategy in large mammals. AB - The substrate demands of lactation must be met by increased dietary intake or by mobilization of nutrients from tissues. The capacity of animals to rely on stored nutrients depends to a large extent on body size; large animals have greater stores, relative to the demands of lactation, than do small animals. The substrate demands of lactation depend on the composition and amount of milk produced. Animals that fast or feed little during lactation are expected to produce milks low in sugar but high in fat, in order to minimize needs for gluconeogenesis while sustaining energy transfers to the young. The patterns of nutrient transfer are reviewed for four taxonomic groups that fast during part of or throughout lactation: sea lions and fur seals (Carnivora: Otariidae), bears (Carnivora: Ursidae), true seals (Carnivora: Phocidae) and baleen whales (Cetacea: Mysticeti). All these groups produce low-sugar high-fat milks, although the length of lactation, rate of milk production and growth of the young are variable. Milk protein concentrations also tend to be low, if considered in relation to milk energy content. Maternal reserves are heavily exploited for milk production in these taxa. The amounts of lipid transferred to the young represent about one-fifth to one-third of maternal lipid stores; the relative amount of the gross energy of the body transferred in the milk is similar. Some seals and bears also transfer up to 16-18 % of the maternal body protein via milk. Reliance on maternal reserves has allowed some large mammals to give birth and lactate at sites and times far removed from food resources. PMID- 10828180 TI - Nutrient partitioning during pregnancy: adverse gestational outcome in overnourished adolescent dams. AB - Appropriate nutrient partitioning between the maternal body and gravid uterus is essential for optimum fetal growth and neonatal survival, and in adult sheep nutrient partitioning during pregnancy generally favours the conceptus at the expense of the dam. However, recent studies using an overnourished adolescent sheep model demonstrate that the hierarchy of nutrient partitioning during pregnancy can be dramatically altered in young growing females. Overnourishing the adolescent dams to promote rapid maternal growth throughout pregnancy results in a major restriction in placental mass and leads to a significant decrease in birth weight relative to moderately-fed adolescents of equivalent gynaecological age. High maternal feed intakes are also associated with an increased incidence of non-infectious spontaneous abortion, a reduction in gestation length and colostrum production, and a higher incidence of neonatal mortality. The present paper examines the putative role of a variety of endocrine regulators of nutrient partitioning in this unusual model system, where the dam is overnourished while the stunted placenta restricts nutrient supply to the fetus. The central role of nutritionally-mediated alterations in placental growth and development in setting the subsequent pattern of nutrient partitioning between the maternal body, placenta and fetus is examined, and critical periods of sensitivity to alterations in maternal nutritional status are defined. Finally, the consequences of this form of inappropriate nutrient partitioning on the growth and development of the fetus and neonate are described with particular emphasis on the reproductive axis. PMID- 10828181 TI - Protein nutrition in late pregnancy, maternal protein reserves and lactation performance in dairy cows. AB - Empirical evidence suggests that prolonged underfeeding of protein to late pregnant dry cows can have modest negative carry-over effects on milk volume and/or protein yield during early lactation, and may also cause increased incidence of metabolic diseases associated with fatty liver. However, assessment of requirements is hampered by lack of information on relationships between dietary intake of crude protein (N x 6.25) and metabolizable protein supply during late pregnancy, and by incomplete understanding of the quantitative metabolism of amino acids in maternal and conceptus tissues. Inability of the postparturient cow to consume sufficient protein to meet mammary and extra mammary amino acid requirements, including a significant demand for hepatic gluconeogenesis, necessitates a substantial, albeit transient, mobilization of tissue protein during the first 2 weeks of lactation. Ultimately, much of this mobilized protein appears to be derived from peripheral tissues, especially skeletal muscle and, to a lesser extent, skin, through suppression of tissue protein synthesis, and possibly increased proteolysis. In the shorter term, soon after calving, it is likely that amino acids required for hepatic glucose synthesis are diverted from high rates of synthesis of splanchnic tissue and export proteins, including serum albumin. The prevailing endocrine milieu of the periparturient cow, including major reductions in plasma levels of insulin and insulin-like growth factor-I, together with insulin resistance in peripheral tissues, must permissively facilitate, if not actively promote, net mobilization of amino acids from these tissues. PMID- 10828182 TI - Adipose tissue metabolism and its role in adaptations to undernutrition in ruminants. AB - Changes in the amount and metabolism of adipose tissue (AT) occur in underfed ruminants, and are amplified during lactation, or in fat animals. The fat depot of the tail of some ovine breeds seems to play a particular role in adaptation to undernutrition; this role could be linked to its smaller adipocytes and high sensitivity to the lipolytic effect of catecholamines. Glucocorticoids and growth hormone probably interact to induce teleophoretic changes in the AT responses to adenosine and catecholamines during lactation. Fat mobilization in dry ewes is related both to body fatness and to energy balance. The in vivo beta-adrenergic lipolytic potential is primarily related to energy balance, whereas basal postprandial plasma non-esterified fatty acids (NEFA) are related to body fatness, and preprandial plasma NEFA is the best predictor of the actual body lipid loss. Several mechanisms seem to be aimed at avoiding excessive fat mobilization and/or insuring a return to the body fatness homeostatic set point. As well as providing the underfed animal with fatty acids as oxidative fuels, AT acts as an endocrine gland. The yield of leptin by ruminant AT is positively related to body fatness, decreased by underfeeding, beta-adrenergic stimulation and short day length, and increased by insulin and glucocorticoids. This finding suggests that the leptin chronic (or acute) decrease in lean (or underfed respectively) ruminants is, as in rodents, a signal for endocrine, metabolic and behavioural adaptations aimed at restoring homeostasis. PMID- 10828183 TI - Malnutrition, morbidity and mortality in children and their mothers. AB - While being underweight or stunted is recognized as an important risk factor for increased prevalence and severity of infection and high mortality rates, there is increasing evidence for an independent role for micronutrient deficiency. Improving vitamin A status reduces mortality among older infants and young children and reduces pregnancy-related mortality; it also reduces the prevalence of severe illness and clinic attendance among children. Improving Zn status reduces morbidity from diarrhoeal and respiratory infection. Treatment of established infection with vitamin A is effective in measles-associated complications, but is not as useful in the majority of diarrhoeal or respiratory syndromes. Zn supplements, however, have significant benefit on the clinical outcome of diarrhoeal and respiratory infections. Concerns that Fe supplements might increase morbidity if given in malarious populations appear to be decreasing, in the light of new studies on Fe supplements showing improved haemoglobin without an increase in morbidity. Breast-feeding, well known to protect against diarrhoea, is also important in protecting against respiratory infection, especially in the young infant. Transmission of human immunodeficiency virus (HIV) in breast milk is recognized, but new data showing reduced transmission in infants who receive exclusive breast-feeding rather than mixed feeding reinforces the importance of promoting this practice in areas where environmental contamination precludes the safe use of other infant feeding regimens. The presence of subclinical mastitis, now recognized to occur in approximately 20 % of mothers in several developing countries, has been shown to increase the concentration of HIV in breast milk. Preliminary findings suggest that the prevalence of subclinical mastitis is reduced by dietary supplements containing antioxidants. Governments and international agencies now have a strong scientific basis to be much more active and innovative in the introduction of focused nutrition interventions especially micronutrients, for the control of infection. PMID- 10828184 TI - The impact of infection and nutrition on gut function and growth in childhood. AB - Poor growth performance during infancy and early childhood is a frequent fact of life in most developing countries. Work in The Gambia has demonstrated that more than 43 % of observed growth faltering during the first 15 months of life can be explained by the presence of a mucosal enteropathy in the small intestine. Within communities the illness is very common: in the area investigated more than 95 % of infants above 8 months of age were affected, and on average they suffered a growth-limiting enteropathy for more than 75 % of their first year of life. Two mechanisms of weight loss have been defined. First, partial villus atrophy reduces absorption and digestion of lactose and probably other nutrients. Second, and more importantly, damage to the mucosal barrier allows translocation of macromolecules into the mucosa and blood, triggering both local and systemic immune and inflammatory mechanisms. Given the severity of the enteropathy it is not surprising that infants fail to grow at a normal rate. Recent findings suggest that these lesions continue throughout childhood and into adulthood. Thus, a persistence of chronic, local and systemic inflammation throughout childhood may be responsible for continued poor growth during this period. Although the nature of the enteropathy and the mechanisms of growth failure have been defined, the factors involved in the initiation and persistence of the intestinal lesion remain uncertain, making clinical management difficult. More work is clearly required to elucidate these factors and to define interventions to prevent or treat the enteropathy. PMID- 10828185 TI - Nutrition in paediatric human immunodeficiency virus infection. AB - The nutritional condition of children with human immunodeficiency virus (HIV) infection continues to be a problem both in developed and developing countries. HIV-infected children grow below normal standards in both height and weight when compared with HIV-exposed non-infected children. These patterns persist over time. It is possible that acute infectious episodes and increased HIV viral burden contribute to decrements in all growth variables. Potential aetiologies for abnormal growth include inadequate dietary intake, gastrointestinal malabsorption, increased energy utilization and psycho-social problems. It is likely that all these factors contribute to the growth problems of these children to some extent. With the development of protease inhibitor anti-retroviral therapy and highly-active anti-retroviral treatment regimens, children with HIV infection in developed countries are living longer with a chronic illness. New nutritional problems have arisen with the development of the fat redistribution syndrome or lipodystrophy. Emerging problems are now being recognized, with the development of insulin resistance and truncal obesity which may potentially lead to premature cardiovascular disease. PMID- 10828186 TI - Water peak suppression: time-frequency vs time-scale approach. AB - Wavelets are the most popular time-scale analysis tool. A well-known application of wavelets in nuclear magnetic resonance spectroscopy is water peak extraction/suppression. However, spectroscopists are more familiar with frequency than scale. So, from a spectroscopist point of view, a time-scale analysis tool (i.e., wavelets) is not natural and a time-frequency approach would be much more satisfactory. We explain a time-frequency solution to this problem based on Gabor analysis. As the two formalisms are closely linked together we continuously emphasize their similarities and differences. In particular we show that, here, the Gabor method is as efficient as the wavelet approach, and we give some examples. Those remarks also apply to other NMR problems solved previously with the continuous wavelet transform, such as quantification or dynamical phase correction. PMID- 10828187 TI - Anisotropy in tendon investigated in vivo by a portable NMR scanner, the NMR MOUSE. AB - Ordered tissue like tendon is known to exhibit the magic-angle phenomenon in magnetic resonance investigations. Due to the anisotropic structure the transverse relaxation time T(2) depends on the orientation of the tendon in the magnetic field. In medical imaging, relaxation measurements of tendon orientation are restricted by the size of the object and the space available in the magnet. For humans, tendon orientation can only be varied within small limits. As a consequence, the magic-angle phenomenon may lead to a misjudgement of tendon condition. It is demonstrated that the NMR-MOUSE (mobile universal surface explorer), a hand-held NMR sensor, can be employed to investigate the anisotropy of T(2) in Achilles tendon in vivo. The NMR-MOUSE provides a convenient tool for analyzing the correlation of T(2) and the physical condition of tendon. PMID- 10828188 TI - One-dimensional imaging with a palm-size probe. AB - A new portable magnetic resonance imaging device was built. Spatially resolved NMR was achieved by placing a gradient coil pair and a Helmholtz pair type radio frequency probe in the gap between two antiparallel polarized permanent magnets. The flat face of the low-field (nu(proton) = 20 MHz) apparatus allowed for the study of arbitrarily large objects in situ. Relaxation time weighted 1D images were achieved over a 15-mm field of view by a single-point spin-echo sequence. A phase encoding time on the order of 200 micros permited the scanning of a wide range of heterogeneous materials. PMID- 10828189 TI - Deuterium nuclear spin-lattice relaxation times and quadrupolar coupling constants in isotopically labeled saccharides. AB - (13)C and (2)H spin-lattice relaxation times have been determined by inversion recovery in a range of site-specific (13)C- and (2)H-labeled saccharides under identical solution conditions, and the data were used to calculate deuterium nuclear quadrupolar coupling constants ((2)H NQCC) at specific sites within cyclic and acyclic forms in solution. (13)C T(1) values ranged from approximately 0.6 to 8.2 s, and (2)H T(1) values ranged from approximately 79 to 450 ms, depending on molecular structure (0.4 M sugar in 5 mM EDTA (disodium salt) in (2)H(2)O-depleted H(2)O, pH 4. 8, 30 degrees C). In addition to providing new information on (13)C and (2)H relaxation behavior of saccharides in solution, the resulting (2)H1 NQCC values reveal a dependency on anomeric configuration within aldopyranose rings, whereas (2)H NQCC values at other ring sites appear less sensitive to configuration at C1. In contrast, (2)H NQCC values at both anomeric and nonanomeric sites within aldofuranose rings appear to be influenced by anomeric configuration. These experimental observations were confirmed by density functional theory (DFT) calculations of (2)H NQCC values in model aldopyranosyl and aldofuranosyl rings. PMID- 10828190 TI - Changes in ADC caused by tensile loading of rabbit achilles tendon: evidence for water transport. AB - Water diffusion measurements were performed on rabbit Achilles tendons during static tensile loading and tendons in an unloaded state. The apparent diffusion coefficient (ADC) was measured along two directions: parallel and perpendicular to the long axis of the tendon. Tendons were studied after being prepared in two ways: (a) after being stored frozen in phosphate-buffered saline (PBS) and (b) freshly isolated. Statistically significant directional anisotropy was observed in the ADC in all tendons. The ADC was significantly greater in the direction parallel to the long axis of the tendon than in the perpendicular direction. The anisotropy is attributed to the greater restrictions seen by the water molecules in the perpendicular direction and is consistent with the known geometry of the tendon. Storage in PBS caused tendons to swell. This increased the ADC measured along both directions and reduced the anisotropy. The existence of anisotropy in the ADC was not related to the orientation of the specimen in the magnet. The ADC increased along both directions following the application of a 5-N tensile load; the increase was greatest along the perpendicular axis of the tendon. In order to determine whether load-related changes in the ADC reflected changes in interfibrilar spacing, we used electron microscopy to measure load-related changes in fibril spacing. Load-related changes in fiber spacing could not account for the observed changes in the ADC. The increase in ADC caused by loading was attributed to the extrusion of tendon water into a bulk phase along the outside surface of the tendon. In PBS-stored samples, enough fluid was extruded that it could be visualized. The transient response of the ADC to a 5-N tensile load was also studied. The absolute ADC in both directions increased with loading and recovered to baseline upon unloading. The transient changes in ADC, for both loading and unloading, had a mean time constant of approximately 15 min. The magnitude of the load-induced transient ADC changes was comparable to that seen in the static-loading experiments. PMID- 10828191 TI - Intensity of cross-peaks in hyscore spectra of S = 1/2, I = 1/2 spin systems. AB - The cross-peak intensity for a S = 1/2, I = 1/2 spin system in two-dimensional HYSCORE spectra of single-crystals and powders is analyzed. There is a fundamental difference between these two cases. For single crystals, the cross peak intensity is distributed between the two (+, +) and (+, -) quadrants of the hyperfine sublevel correlation (HYSCORE) spectrum by the ratio c(2):s(2) (C. Gemperle, G. Aebli, A. Schweiger, and R. R. Ernst, J. Magn. Reson. 88, 241 (1990)). However, for powder spectra another factor becomes dominant and governs cross-peak intensities in the two quadrants. This factor is the phase interference between modulation from different orientations of the paramagnetic species. This can lead to essentially complete disappearance of the cross-peak in one of the two (+, +) or (+, -) quadrants. In the (+, +) quadrant, cross-peaks oriented parallel to the main (positive) diagonal of the HYSCORE spectrum are suppressed, while the opposite is true in the (+, -) quadrant where cross-peaks nearly perpendicular to the main (negative) diagonal of HYSCORE spectra are suppressed. Analytical expressions are derived for the cross-peak intensity profiles in powder HYSCORE spectra for both axial and nonaxial hyperfine interactions (HFI). The intensity is a product of two terms, one depending only on experimental parameter (tau) and the other only on the spin Hamiltonian. This separation provides a rapid way to choose tau for maximum cross-peak intensity in a region of interest in the spectrum. For axial HFI, the Hamiltonian-dependent term has only one maximum and decreases to zero at the canonical orientations. For nonaxial HFI, this term produces three separate ridges which outline the whole powder lineshape. These three ridges have the majority of the intensity in the HYSCORE spectrum. The intensity profile of each ridge resembles that observed for axial HFI. Each ridge defines two principal values of the HFI similar to the ridges from an axial HFI. PMID- 10828192 TI - High-resolution diffusion imaging using a radial turbo-spin-echo sequence: implementation, eddy current compensation, and self-navigation. AB - This work describes a segmented radial turbo-spin-echo technique (DW-rTSE) for high-resolution multislice diffusion-weighted imaging and quantitative ADC mapping. Diffusion-weighted images with an in-plane resolution of 700 microm and almost free of bulk motion can be obtained in vivo without cardiac gating. However, eddy currents and pulsatile brain motion cause severe artifacts when strong diffusion weighting is applied. This work explains in detail the artifacts in projection reconstruction (PR) imaging arising from eddy currents and describes an effective eddy current compensation based on the adjustment of gradient timing. Application of the diffusion gradients in all three orthogonal directions is possible without degradation of the images due to eddy current artifacts, allowing studies of the diffusional anisotropy. Finally, a self navigation approach is proposed to reduce residual nonrigid body motion artifacts. Five healthy volunteers were examined to show the feasibility of this method. PMID- 10828193 TI - Imaging of heterogeneous materials with a turbo spin echo single-point imaging technique. AB - A magnetic resonance imaging method is presented for imaging of heterogeneous broad linewidth materials. This method allows for distortionless relaxation weighted imaging by obtaining multiple phase encoded k-space data points with each RF excitation pulse train. The use of this method, turbo spin echo single point imaging-(turboSPI), leads to decreased imaging times compared to traditional constant-time imaging techniques, as well as the ability to introduce spin-spin relaxation contrast through the use of longer effective echo times. Imaging times in turboSPI are further decreased through the use of low flip angle steady-state excitation. Two-dimensional images of paramagnetic doped agarose phantoms were obtained, demonstrating the contrast and resolution characteristics of the sequence, and a method for both amplitude and phase deconvolution was demonstrated for use in high-resolution turboSPI imaging. Three-dimensional images of a partially water-saturated porous volcanic aggregate (T(2L) approximately 200 ms, Deltanu(1/2) approximately 2500 Hz) contained in a hardened white Portland cement matrix (T(2L) approximately 0.5 ms, Deltanu(1/2) approximately 2500 Hz) and a water-saturated quartz sand (T(2) approximately 300 ms, T(2)(*) approximately 800 microseconds) are shown. PMID- 10828194 TI - Determination of molecular geometry by high-order multiple-quantum evolution in solid-state NMR. AB - The principles of molecular geometry determination by high-quantum heteronuclear local field spectroscopy in solid-state NMR are discussed. The extreme multiple quantum coherences in a cluster of nuclear spins are allowed to evolve in the presence of heteronuclear through-space couplings to two spins of a different type. The multiple-quantum dephasing curve is independent of the homonuclear spin spin couplings and may be described in terms of geometric parameters. The triple quantum version of the experiment is demonstrated by determining the psi torsion angle in a [(15)N(2), (13)C(3)]-labeled sample of the peptide ala-ala-gly. Two regions of torsion angle space fit the experimental data, one in the neighborhood of -152 degrees and one in the neighborhood of +161 degrees. The latter determination is in excellent agreement with the X-ray estimate of +160.5 degrees. PMID- 10828195 TI - Errors in the measurement of cross-correlated relaxation rates and how to avoid them. AB - Cross-correlated relaxation rates Gamma are commonly obtained from constant time experiments by measuring the effect of the desired cross-correlated relaxation on an appropriate coherence during the constant time T. These measurements are affected by systematic errors, which derive from undesired cross-correlated relaxation effects taking place before and after the constant time period T. In this paper we discuss the sources and the size of these errors in an example of two pulse sequences. Higher accuracy of the measured data can be obtained by recording a set of experiments with different T values. Cross-correlated relaxation rates are measured in constant time experiments either from the differential relaxation of multiple components (J-resolved Gamma experiments) or from the efficiency of magnetization transfer between two coherences (quantitative Gamma experiments). In this paper we calculate analytically the statistical errors in both J-resolved and quantitative Gamma experiments. These formulae provide the basis for the choice of the most efficient experimental approach and parameters for a given measurement time and size of the rate. The optimal constant time T for each method can be calculated and depends on the relaxation properties of the molecule under investigation. Moreover, we will show how to optimize the relative duration of cross and reference experiments in a quantitative Gamma approach. PMID- 10828196 TI - Magnetic resonance imaging of electroconvection in a polar organic solvent. AB - Molecular motion in the polar organic solvent nitrobenzene induced by an electric field is studied by magnetic resonance imaging. Rf pulse sequences that correlate images obtained at two different times under conditions of both continuous and pulsed electric fields are introduced. The resultant image correlation spectra indicate that the time scale of motion in a 9.6 kV/cm electric field is tens of milliseconds. Comparison of the results to an analytic solution for the Fokker Planck probability function for one-dimensional bounded diffusion yields an electric field dependent effective diffusion coefficient for perdeuterated nitrobenzene of D = 1.08 x 10(-5) cm(2)/s + (3.33 x 10(-3) cm(4)/kV(2)s) E(2) at room temperature. Characteristics of this electroconvection and its consequences for combining multidimensional nuclear magnetic resonance with electrical orientation are also discussed. PMID- 10828197 TI - 23Na and (1)H NMR microimaging of intact plants. AB - (23)Na NMR microimaging is described to map, for the first time, the sodium distribution in living plants. As an example, the response of 6-day-old seedlings of Ricinus communis to exposure to sodium chloride concentrations from 5 to 300 mM was observed in vivo using (23)Na as well as (1)H NMR microimaging. Experiments were performed at 11.75 T with a double resonant (23)Na-(1)H probehead. The probehead was homebuilt and equipped with a climate chamber. T(1) and T(2) of (23)Na were measured in the cross section of the hypocotyl. Within 85 min (23)Na images with an in-plane resolution of 156 x 156 micrometer were acquired. With this spatial information, the different types of tissue in the hypocotyl can be discerned. The measurement time appears to be short compared to the time scale of sodium uptake and accumulation in the plant so that the kinetics of salt stress can be followed. In conclusion, (23)Na NMR microimaging promises great potential for physiological studies of the consequences of salt stress on the macroscopic level and thus may become a unique tool for characterizing plants with respect to salt tolerance and salt sensitivity. PMID- 10828198 TI - Detection of nuclear resonance signals: modification of the receiver operating characteristics using feedback. AB - The performance of a nuclear resonance detection system can be quantified using binary detection theory. Within this framework, signal averaging increases the probability of a correct detection and decreases the probability of a false alarm by reducing the variance of the noise in the average signal. In conjunction with signal averaging, we propose another method based on feedback control concepts that further improves detection performance. By maximizing the nuclear resonance signal amplitude, feedback raises the probability of correct detection. Furthermore, information generated by the feedback algorithm can be used to reduce the probability of false alarm. We discuss the advantages afforded by feedback that cannot be obtained using signal averaging. As an example, we show how this method is applicable to the detection of explosives using nuclear quadrupole resonance. PMID- 10828199 TI - Adiabatic decoupling sidebands. AB - An analytical solution is given for amplitudes and phases of adiabatic decoupling sidebands as a function of spin inversion time tau. Since all the adiabatic decoupling phases theta(t, tau) refocus at two periods (2T) of the decoupling pulse, the sidebands are located at n/2T rather than at n/T as observed in other decoupling schemes. The real (R(n)(tau)) and imaginary (I(n)(tau)) amplitudes of the sidebands have symmetry R(n)(tau) = R(-n)(tau) and I(n)(tau) = -I(-n)(tau), forming a mirror image between the counterparts of the sidebands. When frequency sweep changes direction all I(n)(tau) are inverted while all R(n)(tau) remain unchanged, leading to pure absorption sidebands with two accumulations as demonstrated by Kupce and Freeman, and to an exchange of sidebands between counterparts. The sum of the real parts for sidebands n = 1 and 2 is almost a constant near on-resonance decoupling, and it increases substantially for large decoupling offsets. The phase defocusing can be minimized for all decoupling offsets by inserting an initial decoupling period with T(ini) = T/2, eliminating all sidebands located at n/2T (n = +/-1, +/-3, +/-5, ...). PMID- 10828200 TI - ESR spectroscopy as a tool for identifying joining fragments of antique marbles: the example of a pulpit by Donatello and Michelozzo. AB - ESR spectroscopy is one of the physicochemical techniques used to characterize archaeological white marbles and obtain information about their quarries of provenance. This is done by measuring selected spectral features of the Mn(2+) impurity ubiquitously present in marbles and developing a statistical classification rule from the variable vectors measured for a significant number of samples of known provenance (the quarry database). Now we show that the overall variability exhibited by the same spectroscopic features decreases rapidly with the linear dimensions of the sampled block and can be used to distinguish fragments belonging to the same piece of stone from those simply originating from the same quarry. Application of the method to the seven marble panels of the Donatello pulpit in Prato (Tuscany) shows that they have all been cut from the same single block and their different degradation must be ascribed to differential weathering and to the different conservation treatments undergone in the past. The limits and possible drawbacks of the method are also discussed. PMID- 10828201 TI - Multidimensional dipolar exchange-assisted recoupling measurements in solid-state NMR. AB - Aseries of uni- and multidimensional variants of the dipolar exchange-assisted recoupling (DEAR) NMR experiment is described and applied to determinations of (13)C-(14)N dipolar local field spectra in amino acids and dipeptides. The DEAR protocol recouples nearby nuclei by relying on differences in their relative rates of longitudinal relaxation, and has the potential to give quantitative geometric results without requiring radiofrequency pulsing on both members of a coupled spin pair. One- and two-dimensional variants of this recoupling strategy on generic I-S pairs are discussed, and measurements of (13)C-(14)N distances and 2D local field experiments sensitive to the relative orientation of CN vectors with respect to the (13)C shielding tensor are presented. Since these measurements did not involve pulsing on the broad nitrogen resonance, their results were independent of the quadrupolar parameters of this nucleus. High resolution 3D NMR versions of the 2D experiments were also implemented in order to separate their resulting local field patterns according to the isotropic shifts of inequivalent (13)C sites. These high-resolution 3D acquisitions involved collecting a series of 2D DEAR NMR data sets on rotating samples as a function of spinning angle, and then subjecting the resulting data to a processing akin to that involved in variable-angle correlation NMR. Once successfully tested on l-alanine this experiment was applied to the analysis of a series of dipeptides, allowing us to extract separate local field (13)C-(14)N spectra from this type of multisite systems. PMID- 10828202 TI - The multidimensional filter diagonalization method. AB - The theory and numerical aspects of the recently developed multidimensional version of the filter diagonalization method (FDM) are described in detail. FDM can construct various "ersatz" or "hybrid" spectra from multidimensional time signals. Spectral resolution is not limited by the time-frequency uncertainty principle in each separate frequency dimension, but rather by the total joint information content of the signal, i.e., N(total) = N(1) x N(2) x vertical ellipsis x N(D), where some of the interferometric dimensions do not have to be represented by more than a few (e.g., two) time increments. It is shown that FDM can be used to compute various reduced-dimensionality projections of a high dimensional spectrum directly, i.e., avoiding construction of the latter. A subsequent paper (J. Magn. Reson. 144, 357-366 (2000)) is concerned with applications of the method to 2D, 3D, and 4D NMR experiments. PMID- 10828203 TI - The multidimensional filter diagonalization method. AB - The theory of the multidimensional filter diagonalization method (FDM) described in the previous paper (V. A. Mandelshtam, 2000, J. Magn. Reson. 144, 343-356 (2000)) is applied to NMR time signals with up to four independent time variables. Direct projections of the multidimensional time signals produce new kinds of 2D spectra. The resolution obtained by FDM can be far superior to that obtained by conventional phase-sensitive FT processing, and correlation peaks in heteronuclear and homonuclear experiments can be condensed to sharp singlets, removing all spin-spin couplings. Examples of singlet-HSQC and singlet-TOCSY spectra show big gains in resolution. It is not necessary to have a finely digitized spectrum, in which the individual multiplet components are resolved, for the methods to work. Examples of FDM spectra, ranging from simple organic molecules and steroids to metalloproteins, are shown. PMID- 10828204 TI - Protein backbone (15)N relaxation rates as a tool for the diagnosis of structure quality. AB - In the work reported herein we define a structure validation factor that depends on protein backbone (15)N relaxation rates. This is an alternative method to the previously defined quality factors derived from anisotropic chemical shifts or residual dipolar couplings. We have used the structure dependence of (15)N relaxation rates of anisotropically tumbling proteins to calculate this structure diagnosis factor and have used it to demonstrate the improvement of protein structures refined with residual dipolar couplings. PMID- 10828205 TI - Chemical shift imaging with continuously flowing hyperpolarized xenon for the characterization of materials. AB - In this contribution we report new approaches to the MRI of materials using continuously produced laser-polarized (129)Xe gas. This leads to vastly improved sensitivity and makes new kinds of information available. The hyperpolarized xenon is produced in a continuous flow system that conveniently delivers the xenon at low partial pressure to probes for NMR and MRI experiments. We illustrate applications to the study of micropore and other kinds of void space and show for the first time that with flowing hyperpolarized xenon it is possible to obtain chemical-shift-resolved images in a relatively short time. PMID- 10828206 TI - Acute effects of an insect repellent, N,N-diethyl-m-toluamide, on cholinesterase inhibition induced by pyridostigmine bromide in rats. AB - Acute lethal interactions have been previously described between a cholinesterase (ChE) inhibitor, pyridostigmine bromide (PB), and the insect repellent, N,N diethyl-m-toluamide (DEET). The mechanism of toxic interaction between these agents is unknown. Alterations in membrane permeability caused by DEET could facilitate or enhance absorption, or alter the distribution of peripherally restricted PB, causing increased inhibition of ChE at a given dose. Studies were conducted to investigate PB-induced ChE inhibition in the presence of DEET. Rats received ip injections of PB (1, 2, or 3 mg/kg), DEET (200 mg/kg), or PB + DEET at doses that potentiated acute lethality. ChE activity was measured in heart, diaphragm, blood, whole brain, or specific brain areas using a modified spectrophotometric assay. DEET did not alter PB-induced inhibition of ChE activity in rat diaphragm, heart, or blood. Administration of DEET alone had no effect on ChE activity. PB alone did not inhibit ChE in whole brain, but PB (3 mg/kg) + DEET (200 mg/kg) caused significant inhibition of whole brain ChE activity to approximately 60% of controls. In specific brain areas, (cortex, cerebellum, medulla, hypothalamus, hippocampus, midbrain, and striatum) PB alone did not inhibit ChE activity. PB (3 mg/kg) + DEET (200 mg/kg) reduced ChE activity to approximately 65-75% of controls in each brain area, but those results were not statistically significant. In conclusion, DEET did not alter PB induced inhibition of ChE activity in the periphery. While DEET may have facilitated the access of PB into the CNS at high doses, it is doubtful that the resulting minor reduction in ChE activity would have resulted in death. It is unlikely that the lethal interaction between PB and DEET is mediated through a cholinergic effect resulting from increased inhibition of ChE. PMID- 10828207 TI - EGF receptors of hepatocytes from rats treated with phenobarbital are sensitized to down-regulation by phenobarbital in culture. AB - Hepatocytes from Fisher 344 rats treated with the liver tumor promoter phenobarbital (PhB; 0.1% in the drinking water, 2-3 months) exhibit reduced epidermal growth factor (EGF) binding and EGF-induced mitogenesis in culture. Similar responses are induced by >1 mM PhB added to the culture medium of hepatocytes from untreated rats. In this study, we demonstrated that hepatocyte EGFr protein, as determined by immunoblotting, was unchanged by treatment of rats with PhB. However, hepatocytes from PhB-treated rats are more sensitive to PhB in culture in that decreased EGF binding occurred with 0.05 mM PhB, a concentration also attained in plasma of rats exposed to PhB. Sensitization was reversible, as is tumor promotion, since hepatocytes from rats withdrawn from PhB for 1 month were unresponsive to <3 mM PhB. EGFr down-regulation by a series of barbiturates correlated well with their known activities as tumor promoters and CYP2B1/2 inducers, with pentobarbital and PhB yielding high activities, while barbital was intermediate and barbituric acid, 5-phenylbarbituric acid, and 5-ethylbarbituric acid were ineffective. Differentiated hepatocyte function was required for PhB induced EGFr down-regulation since HepG2 and rat liver epithelial cells were unresponsive, but involvement of CYP2B1/2 activity was discounted by the failure of metyrapone to inhibit the response in PhB-induced hepatocytes. These studies support a role for impaired EGFr function in PhB liver tumor promotion due to effects on existing EGFr protein and suggest that EGFr down-regulation by PhB in culture is independent of CYP2B1/2 activity but shares mechanistic components involved in its transcriptional activation by PhB. PMID- 10828208 TI - gamma-diketone peripheral neuropathy. I. Quality morphometric analyses of axonal atrophy and swelling. AB - Quantitative morphometric analysis was used to characterize expression of myelinated axon swelling and atrophy in rat peripheral nerve during 2,5 hexanedione (HD) intoxication. HD was administered by gavage according to different daily dosing regiments (100, 175, 250, or 400 mg/kg/day) and four proximodistal nerve regions (5th lumbar spinal nerve, proximal and distal sciatic nerve, and tibial nerve) were examined morphometrically. Morphometric determinations were made at four behavioral endpoints (unaffected, slight, moderate, and severe toxicity) and were correlated to electrophysiologic measurements of peripheral nerve function. Results show that, for all HD dose rates, onsets of behavioral neurotoxicity and nerve dysfunction were generally related to development of abundant axon atrophy. The proximodistal manifestation of atrophy was dependent upon the dosing rate; i.e., the atrophy response produced by subacute intoxication with higher daily dosing rates (250 and 400 mg/kg/day) was restricted to distal nerve regions whereas subchronic induction with lower dosing rates (100 and 175 mg/kg/day) produced abundant fiber atrophy in all proximodistal areas. In contrast to atrophy, axonal swellings constituted an inconsistent minor morphologic response, the expression of which was dependent upon subchronic dosing rates (100-250 mg/kg/day). Subacute HD administration (400 mg/kg/day) produced significant changes in neurobehavior and nerve electrophysiologic parameters in the absence of peripheral axon swelling. Thus, conditional expression of swellings suggests they are an epiphenomenon related to low-dose induction rates. Fiber atrophy, however, was numerically dominant, correlated with nerve dysfunction, and occurred at all dosing levels. These characteristics suggest atrophy is a neurotoxicologically significant feature of gamma-diketone peripheral neuropathy. PMID- 10828209 TI - gamma-diketone peripheral neuropathy. II. Neurofilament subunit content. AB - Quantitative morphometric analyses have demonstrated that axon atrophy is the primary neuropathic alteration in peripheral nerve of 2,5-hexanedione (HD) intoxicated rats (Lehning et al., Toxicol. Appl. Pharmacol. 165, 127-140, 2000). Research suggests that axon caliber is regulated by neurofilament (NF) content and density. Therefore, as a possible mechanism of atrophy, NF subunit (NF-L, -M, and -H) proteins were quantitated in moderately affected rats intoxicated with HD at three daily dosing rates (175, 250, and 400 mg/kg/day). Analyses of subunit protein contents in proximal sciatic nerves indicated uniformly small decreases, which corresponded to minimal changes in axon area occurring in this region. In distal tibial nerve, subunit proteins were decreased substantially (40-70%) when rats were exposed to the 175 and 250 mg/kg/day doses. These reductions in NFs corresponded to significant decreases (approximately 50%) in tibial axon area induced by lower dosing rates. In contrast, 400 mg/kg/day produced similar changes in caliber but smaller reductions (18-25%) in NF-L, -M, and -H levels. This suggests that a decrement in axonal NF content is unlikely to be solely responsible for gamma-diketone-induced axon atrophy and that the corresponding mechanism probably involves additional changes in factors regulating NF density. Analysis of NF content in peripheral nerve also identified the presence of anomolous higher molecular weight NF-H proteins. However, the neurotoxicological significance of these abnormal subunits is uncertain based on their limited occurrence and inconsistent spatiotemporal expression. PMID- 10828210 TI - Paternal effects from methamidophos administration in mice. AB - In this study, the mouse was used to evaluate paternal germline exposure to the organophosphate methamidophos for its potential to produce adverse effects on spermatozoa and in the offspring. There have been reports that organophosphate exposure can increase abnormal sperm morphology in mice. However, effects transmitted to the offspring following paternal exposure have not been reported previously. The maximum tolerated dose (MTD) was 7.5 mg kg(-1) body weight and this dose resulted in no deaths, although blood plasma cholinesterase activity was still decreased. Males were euthanized 4 weeks after an acute intraperitoneal injection of methamidophos (0.5, 3.75, 5.0, and 7.5 mg kg(-1) body wt) and the number of spermatids per gram testes and sperm morphology were analyzed. In this study, abnormal sperm morphology on a per group basis exhibited a dose-response significantly related to increased methamidophos exposure as indicated by regression analysis and a nested ANOVA (p < 0.0001). Preimplantation embryos that were conceived 6 weeks after paternal methamidophos exposure (5 mg kg(-1) body wt) exhibited a significant increase in cleavage arrest. Fertility of males was also affected as shown by a decrease in the number of two- to four-cell embryos per male (postexposure week 6) and an increase in the number of degenerated embryos (postexposure weeks 4-6). We conclude that methamidophos may have the potential to produce transmissible adverse embryonic effects following an acute paternal germline exposure. PMID- 10828211 TI - Expression of biotransformation enzymes in human fetal olfactory mucosa: potential roles in developmental toxicity. AB - High levels of cytochrome P450 are present in the olfactory mucosa (OM) in mammalian animals and contribute to the known tissue-selective toxicity of numerous chemical compounds. Olfactory toxicity in the perinatal period may have a greater impact on behavior, growth, and development than in adults. To establish a molecular basis for determining the risk of developmental toxicity in OM, the expression of several cytochrome P450 enzymes, as well as NADPH cytochrome P450 reductase and microsomal epoxide hydrolase, was examined in hepatic and nasal microsomes prepared from human fetal tissues at gestational day 91-125. The relative microsomal concentrations of these biotransformation enzymes were determined on immunoblots. Expression of CYP2A, CYP2J2, the reductase, and epoxide hydrolase was detected in both OM and liver. The microsomal levels of these enzymes were generally lower in OM than in liver of the same fetuses, except for the CYP2A-related proteins, which were expressed in OM at much higher levels. OM expression of CYP2A6, CYP2A13, CYP2B6, and CYP2J2 mRNAs was detected using RNA-PCR. These results document, for the first time, prenatal expression of xenobiotic-bioactivating cytochrome P450 enzymes in human OM and suggest that the human fetal OM may be a preferred target tissue for the toxicity of maternally derived chemical compounds that are activated by the CYP2A enzymes. PMID- 10828212 TI - Development of a physiologically based pharmacokinetic model for hydroquinone. AB - Hydroquinone (HQ) produces nephrotoxicity and renal tubular adenomas in male F344 rats following 2 years of oral dosing. Female F344 and SD rats are comparatively resistant to these effects. Nephrotoxicity and tumorigenicity have been associated with a minor glutathione conjugation pathway following the oxidation of HQ to benzoquinone (BQ). The majority of administered doses (90-99%) consists of glucuronide and sulfate conjugates of HQ. An initial physiologically based pharmacokinetic model was developed to characterize the role of kinetics in the strain differences observed in HQ-induced renal toxicity and tumorigenicity. Partition coefficients, protein-binding, and metabolic rate constants were determined directly or estimated from a series of in vivo and in vitro studies. Metabolism was confined to the liver and GI tract. The total flux through the glutathione pathway represented the "internal dose" of HQ for nephrotoxicity. Simulations were compared to a variety of data from male and female F344 rats, male SD rats, and a single male human volunteer. Simulations of intraperitoneal administration resulted in higher amounts of glutathione conjugates than comparable oral doses. This was consistent with protein-binding and toxicity studies and emphasized the importance of first-pass GI tract metabolism. In addition, male F344 rats were predicted to form more total glutathione conjugates than SD rats at equivalent dose levels, which was also consistent with the observed strain differences in renal toxicity. This model represents the first stage in the development of a biologically based dose-response model for improving the scientific basis for human health risk assessments of HQ. PMID- 10828213 TI - Phenogenetic drift and the evolution of genotype-phenotype relationships. PMID- 10828214 TI - Stochastic modelling of environmental variation for biological populations. AB - We examine stochastic effects, in particular environmental variability, in population models of biological systems. Some simple models of environmental stochasticity are suggested, and we demonstrate a number of analytic approximations and simulation-based approaches that can usefully be applied to them. Initially, these techniques, including moment-closure approximations and local linearization, are explored in the context of a simple and relatively tractable process. Our presentation seeks to introduce these techniques to a broad-based audience of applied modellers. Therefore, as a test case, we study a natural stochastic formulation of a non-linear deterministic model for nematode infections in ruminants, proposed by Roberts and Grenfell (1991). This system is particularly suitable for our purposes, since it captures the essence of more complicated formulations of parasite demography and herd immunity found in the literature. We explore two modes of behaviour. In the endemic regime the stochastic dynamic fluctuates widely around the non-zero fixed points of the deterministic model. Enhancement of these fluctuations in the presence of environmental stochasticity can lead to extinction events. Using a simple model of environmental fluctuations we show that the magnitude of this system response reflects not only the variance of environmental noise, but also its autocorrelation structure. In the managed regime host-replacement is modelled via periodic perturbation of the population variables. In the absence of environmental variation stochastic effects are negligible, and we examine the system response to a realistic environmental perturbation based on the effect of micro-climatic fluctuations on the contact rate. The resultant stochastic effects and the relevance of analytic approximations based on simple models of environmental stochasticity are discussed. PMID- 10828215 TI - Population dynamics of plant-parasite interactions: thresholds for invasion. AB - Thresholds are derived for the invasion of plant populations by parasites. The theory is developed for a generic model that takes into account two features characteristic of plant-parasite interactions: a dual source of inoculum (infection from primary or externally introduced inoculum and secondary infection from contact between susceptible and infected host tissue) and a host response to infection load. Each of the threshold criteria is shown to be the sum of the individual components for primary and secondary infection. This indicates that if parasite invasion is not possible through primary or secondary infection alone, when the two modes of transmission are combined, the parasite may be able to invade. The invasion criteria demonstrate that there is a threshold population of susceptible hosts below which the parasite is unable to invade. If there are nonlinearities in the population dynamics (arising through either the transmission process or the host response), there are also threshold densities for the infected hosts and parasite populations below which invasion does not occur. The implications of the results for the control of plant disease are discussed. PMID- 10828216 TI - A model for tuberculosis with exogenous reinfection. AB - Following primary tuberculosis (TB) infection, only approximately 10% of individuals develop active T.B. Most people are assumed to mount an effective immune response to the initial infection that limits proliferation of the bacilli and leads to long-lasting partial immunity both to further infection and to reactivation of latent bacilli remaining from the original infection. Infected individuals may develop active TB as a consequence of exogenous reinfection, i.e., acquiring a new infection from another infectious individual. Our results in this paper suggest that exogenous reinfection has a drastic effect on the qualitative dynamics of TB. The incorporation of exogenous reinfection into our TB model allows the possibility of a subcritical bifurcation at the critical value of the basic reproductive number R(0)=1, and hence the existence of multiple endemic equilibria for R(0)<1 and the exogenous reinfection rate larger than a threshold. Our results suggest that reducing R(0) to be smaller than one may not be sufficient to eradicate the disease. An additional reduction in reinfection rate may be required. These results may also partially explain the recently observed resurgence of TB. PMID- 10828217 TI - The stability of symmetric solutions to polygenic models. AB - Analysis of multilocus evolution is usually intractable for more than n approximately 10 genes, because the frequencies of very large numbers of genotypes must be followed. An exact analysis of up to n approximately 100 loci is feasible for a symmetrical model, in which a set of unlinked loci segregate for two alleles (labeled "0" and "1") with interchangeable effects on fitness. All haploid genotypes with the same number of 1 alleles can then remain equally frequent. However, such a symmetrical solution may be unstable: for example, under stabilizing selection, populations tend to fix any one genotype which approaches the optimum. Here, we show how the 2(n)x2(n) stability matrix can be decomposed into a set of matrices, each no larger than nxn. This allows the stability of symmetrical solutions to be determined. We apply the method to stabilizing and disruptive selection in a single deme and to selection against heterozygotes in a linear cline. PMID- 10828218 TI - On estimating the heterozygosity and polymorphism information content value. AB - The polymorphism information content (PIC) value is commonly used in genetics as a measure of polymorphism for a marker locus used in linkage analysis. In this communication we have derived the uniformly minimum variance unbiased estimator of PIC along with its exact variance. We have also calculated the exact variance of the maximum likelihood estimator of PIC which is asymptotically an unbiased estimator. In order to find this variance we have derived a recursive formula to calculate the moments of any polynomial in a set of variables that are multinomially distributed. PMID- 10828219 TI - Species coexistence by permanent spatial heterogeneity in a lottery model. AB - We studied the effect of permanent spatial heterogeneity in promoting species coexistence in a lottery model. The system consisted of multiple habitats, each composed of a number of sites occupied by adults of two species. Larvae produced from different habitats were mixed in a common pool. When an adult died, the vacant site became occupied by an individual randomly chosen from the larval pool. If there were n habitats, there could be up to n-1 internal equilibria with both species in addition to two single-species equilibria. These equilibria and their local stability can be calculated from a single function, indicating the difference among species in their average lifetime reproductive success. Our main result is that between-habitat variation in the ratio of mortalities of two species promotes coexistence, while that of reproductive rates does not. This conclusion is the opposite of the role of temporal variation in the standard lottery model, in which between-year variation in the reproductive rate, but not that in the mortalities, promotes coexistence. PMID- 10828220 TI - Properties of bias and variance of two multiallelic estimators of F(ST). AB - This study compares theoretical and simulated properties of two estimators of fixation indices F(ST) for multiallelic data from a single locus. The estimators are due to Weir and Cockerham (straight theta(WC)) and to Robertson and Hill (;straight theta(RH)), respectively. Both estimators are linear combinations of biallelic estimators which differ in the way frequent and rare alleles are weighted. Consequently, their sampling properties differ as far as bias and variance are concerned. In the infinite island model at migration-drift equilibrium, in the case of one multiallelic locus, we analytically approximate the bias of the two estimators and show that;straight theta(WC) is nearly unbiased, whereas;straight theta(RH) is negatively biased. Hence, we propose a correction of bias of the latter. Moreover, we reexamine the properties of variance of the initial estimators: due to their construction, their variances are minimal, each over different parameter ranges,;straight theta(RH) performing better for low differentiation and;straight theta(WC) for high differentiation. These theoretical properties are confirmed by simulations, which show that our correction of bias of;straight theta(RH) performs well and does not affect its property of minimal variance for low differentiation. Hence, we advocate the use of;straight theta(WC) for high values of differentiation, and the use of;straight theta(RH), with our correction of bias, for low or moderate differentiation. PMID- 10828221 TI - Quasi-equilibrium theory for the distribution of rare alleles in a subdivided population: justification and implications. AB - This paper examines a quasi-equilibrium theory of rare alleles for subdivided populations that follow an island-model version of the Wright-Fisher model of evolution. All mutations are assumed to create new alleles. We present four results: (1) conditions for the theory to apply are formally established using properties of the moments of the binomial distribution; (2) approximations currently in the literature can be replaced with exact results that are in better agreement with our simulations; (3) a modified maximum likelihood estimator of migration rate exhibits the same good performance on island-model data or on data simulated from the multinomial mixed with the Dirichlet distribution, and (4) a connection between the rare-allele method and the Ewens Sampling Formula for the infinite-allele mutation model is made. This introduces a new and simpler proof for the expected number of alleles implied by the Ewens Sampling Formula. PMID- 10828222 TI - Antithrombotic agents after peripheral transluminal angioplasty: a review of the studies, methods and evidence for their use. AB - OBJECTIVE: to review published studies and assess the strength of the evidence for the use of antithrombotic agents for the prevention of re-occlusion after peripheral angioplasty. METHODS: a literature search was performed. All randomised comparative studies with patency assessments or clinical endpoints were included. The methods and results of the studies were compared and evaluated. RESULTS: eleven randomised trials were identified, six of them were double blind. The majority of patients included were those with femoropopliteal lesions. A significant benefit of aspirin in one placebo-controlled study is balanced by no apparent benefit in another study. No dose-dependent effect of aspirin in the range 50-1000 mg/day has been shown in any of the four studies investigating such an effect. None of the three studies comparing platelet inhibitors and oral anticoagulants have shown any differences in outcome and no other well-designed studies of anticoagulants have been reported. CONCLUSIONS: evidence for a reduction in the likelihood of re-occlusion or restenosis after peripheral transluminal angioplasty with platelet inhibitors remains equivocal. Evidence for the efficacy of any other agent in this indication is also lacking. PMID- 10828223 TI - A randomised controlled trial of Prostar Plus for haemostasis in patients after coronary angioplasty. AB - OBJECTIVES: to clarify the efficacy and safety of Prostar Plus, a new percutaneous vascular surgical device (PVS) for vascular haemostasis. DESIGN: prospective randomised controlled trial. METHODS: a consecutive series of 60 patients were randomised to either PVS (n =30) or conventional manual compression ( n =30) following coronary angioplasty or stenting with femoral access using an 8-F sheath. RESULTS: PVS significantly shortened the time to haemostasis (10 s.d. 3 vs. 27 s.d. 9 min, p <0.001), ambulation (2.2 s.d. 0.9 vs. 11.0 s.d. 1.4 h, p <0.001), and discharge (2.2 s.d. 0.4 vs. 3.1 s.d. 0.7 days, p <0.01), compared with the manual compression group with no major complications. PVS also increased patient comfort assessed by using a visual-analogue scale method. Although these clinical benefits reduced the hospital cost ($1301 s. d. 248 vs. 1613 s.d. 460, p <0.05), the cost of the PVS device (approximately $350) cancelled the cost-saving benefit. CONCLUSIONS: this randomised study indicates that Prostar Plus is safe, more effective and comfortable than conventional manual compression. PMID- 10828224 TI - Prevention of intimal hyperplasia by single-dose pre-insertion external radiation in canine-vein interposition grafts. AB - OBJECTIVES: to evaluate the efficacy of single-dose pre-insertion gamma radiation of vein grafts in the prevention of intimal hyperplasia. METHODS: femoral artery interposition grafts with internal jugular vein were inserted in 12 mongrel dogs. The animals were randomly divided into two groups. Immediately before graft replacement, jugular veins were treated with a single dose of cobalt-60 radiation at 14 Gy or received no radiation (control group). Six weeks after graft insertion, the vein grafts were pressure-perfusion fixed and harvested for the histomorphometric analysis. Quantitative data on anastomotic stenosis were calculated from Gilman parameters after cross-sectional image analysis. RESULTS: vein grafts treated with radiation demonstrated significantly decreased neointima formation compared with grafts in the control group. The mean Gilman parameter for the control group was 1.09 S.E.M. 0.34 mm and for the radiotherapy group was 0.65 S.E. M. 0.23 mm (p<0.05). All vein grafts in the radiotherapy group had a decreased amount of intimal and cellular infiltration. CONCLUSION: single-dose external pre-insertion gamma radiation of vein grafts reduced the amount of intimal hyperplasia in this animal model. PMID- 10828225 TI - Marimastat inhibits neointimal thickening in a model of human arterial intimal hyperplasia. AB - OBJECTIVE: matrix metalloproteases (MMPs) produced by vascular smooth-muscle cells (VSMCs) degrade extracellular matrix and facilitate the migration of these cells. This is a fundamental process in arterial intimal hyperplasia. This study investigated whether Marimastat (a selective but non-specific MMP inhibitor) can prevent intimal hyperplasia in cultured human internal mammary artery (IMA). MATERIALS AND METHODS: segments of IMA from 8 patients were prepared and cultured for 14 days in serum-supplemented medium (control) or in medium supplemented with Marimastat at 2 concentrations (treatment groups). The tissue was fixed, sectioned, stained and neointimal thicknesses measured by computer-aided image analysis. Further sections were cultured in the same manner and prepared for gel enzymography to quantify the production of MMPs. RESULTS: neointimal thickness was significantly reduced by Marimastat in a dose-dependent manner when compared to controls (p =0.008 Wilcoxon). Gel enzymography demonstrated a reduction in levels of MMP2 and MMP9. This was most significant for the active forms of the enzymes ( p =0.03). CONCLUSIONS: our results suggest that there is a potential therapeutic role for specific inhibition of the gelatinases in the prevention of human arterial restenosis. PMID- 10828226 TI - Exposure of plasma proteins on Dacron and ePTFE vascular graft material in a perfusion model. AB - OBJECTIVES: to compare the exposure of plasma proteins adsorbed onto three vascular graft materials (polytetrafluoroethylene ePTFE and two modifications of polyethyleneterephthalate Dacron). METHODS: surface exposure of fibronectin, vitronectin, thrombospondin, antithrombin III, IgG, high molecular-weight kininogen, fibrinogen, albumin and plasminogen was studied by incubation with radiolabelled antibodies in a perfusion model. Perfusion times with human plasma were 1, 4, 24 and 48 hours. RESULTS: all proteins could be detected at 1, 4, 24 and 48 hours after the start of perfusion. Overall, the least amount of proteins adsorbed onto ePTFE. CONCLUSIONS: the low adsorption of proteins onto ePTFE may be one of the reasons for the lower incidence of infections reported with this material. PMID- 10828227 TI - Above-knee prosthetic femoropopliteal bypass for intermittent claudication. Results of the initial and secondary procedures. AB - OBJECTIVES: [corrected] to report the results of primary and secondary prosthetic above-knee femoropopliteal bypass for intermittent claudication. DESIGN: a retrospective study in a University hospital. PATIENTS: one hundred and twelve operations performed in 103 patients (26 women) between January 1990 and June 1997. METHODS: a comparison of primary assisted patency was made between Dacron and PTFE, between men and women and between operations performed early and late in the study period. Patency of secondary procedures was also studied. RESULTS: there were no operative deaths. The 5-year survival rate was 81% and equal to that of a demographically matched population. The primary assisted graft patency was 58% after two years. Women had a significantly better graft patency than men (79% vs. 49%). The type of graft and the date of the operation did not influence the outcome. Forty of the 55 occluded grafts were subjected to a redo procedure with a 1-year patency of 29%. CONCLUSIONS: the results after prosthetic above knee femoropopliteal bypass procedures are disappointing, and a controversy persists as to whether this operation should be performed for intermittent claudication. The results of secondary procedures are even worse, and perhaps should only be considered in patients suffering critical ischaemia. PMID- 10828228 TI - Endoluminal femoropopliteal bypass for intermittent claudication. AB - OBJECTIVES: (i) to describe our initial clinical experience with endoluminal femoropopliteal bypass using a technique developed in a cadaveric model; (ii) to identify areas requiring technical modification to improve patency and complication rates. DESIGN: prospective, experimental pilot study. MATERIALS AND METHODS: fourteen consecutive patients with disabling intermittent claudication and superficial femoral artery occlusion underwent endarterectomy through a groin incision and endoluminal placement of a polytetrafluoroethylene graft. Follow-up was by duplex ultrasound and arteriography. RESULTS: two endovascular technical failures required conversion to open surgery. The cumulative primary (1 degrees), 1 degrees-assisted and secondary (2 degrees) patency rates at 1 year were 35.7%, 42.8% and 71.4% respectively; at 2 years the patency rates were 14.3%, 31.2% and 57.1%. Twenty-three endovascular interventions were required to maintain graft patency in 10 patients. Five patients subsequently required conventional bypass, of whom two proceeded to major amputation because of graft infection. Seven endovascular grafts remain patent at a mean follow-up of 50 months. CONCLUSIONS: this minimally invasive surgical technique is feasible, with acceptable patency rates. However, considerable investment of time and resources is required to maintain graft patency. With increasing experience and improved technical design, this procedure may offer a real alternative to conventional surgery in patients disabled by short-distance claudication. PMID- 10828229 TI - Endovascular treatment of penetrating thoracic outlet arterial injuries. AB - OBJECTIVES: to establish the feasibility of stent-graft treatment of penetrating thoracic outlet arterial injuries. DESIGN: prospective study. MATERIALS AND METHODS: forty-one patients with penetrating injuries to the carotid, subclavian and proximal axillary arteries admitted between August 1998 and May 1999 were studied. Patients requiring urgent surgical exploration for active bleeding (n=26) were excluded. Remaining patients underwent arteriography to assess suitability for stent-graft placement. After successful stent-graft treatment clinical and sonographic follow-up were done at 1 month and thereafter 3-monthly. RESULTS: of the 15 patients considered, 10 patients qualified for stent-graft treatment (seven male, three female, mean age 27 years). The vessels involved were subclavian artery (seven), carotid artery (two) and axillary artery (one). Seven had arteriovenous fistulae and three, pseudoaneurysms. Stent-graft treatment was successful in all 10 patients with no procedure-related complications. On mean follow-up of 7 months no complications were encountered. CONCLUSION: endovascular treatment shows promise as a treatment modality for thoracic outlet arterial injuries. Long-term follow-up is required for comparison to the results of standard surgical repair. PMID- 10828230 TI - Limb salvage and popliteal aneurysms: advantages of preventive surgery. AB - OBJECTIVES: to assess the prognosis of atherosclerotic popliteal aneurysms (APAs), according to whether they were occluded or patent at the time of diagnosis. DESIGN: retrospective study. PATIENTS AND METHODS: fifty-two APAs were investigated in 35 patients. Nineteen were occluded (group I) and 33 patent (group II). In group I, 11 lower limbs had critical ischaemia, and eight had severe claudication. In group II, 27 were asymptomatic, 3 were painful, and 3 presented with symptomatic distal occlusion. In group I, treatment consisted of six bypasses, five thrombectomies, four thrombolyses, but for five APAs, no revascularisation was possible due to lack of runoff. In group II, 30/33 APAs were treated by graft replacement; the other three were not operated on due to the patients>> poor general condition. RESULTS: the 4-year survival rate was 72% in group I vs. 77% in group II, and the limb salvage rate was 72% in group I vs. 100% in group II, p<0.01. CONCLUSION: prophylactic treatment of asymptomatic popliteal aneurysms may avoid amputation caused by thrombosis and embolisation of runoff. PMID- 10828231 TI - Intravascular ultrasound underestimates vessel dimensions. AB - The information gathered with intravascular ultrasound (IVUS) are of great value in endovascular techniques. The aim of this study was to evaluate the reliability of IVUS when measuring vessel dimensions by comparison with an established reference method. The left carotid artery was exposed in 4 pigs (45-55 kg) and two piezoelectric crystals were sutured on the adventitia in the same cross sectional plane. The distance between them was measured either by IVUS and by sonomicrometers. The mean distance between the two crystals calculated by the sonomicrometer was 4.7+/-0.4 mm (mean systolic distance was 4.9+/-0.2 mm, mean diastolic distance was 4.6+/-0.1 mm). The mean distance between the two targets calculated by IVUS was 4. 5+/-0.2 mm (mean systolic distance was 4.6+/-0.2 mm and mean diastolic 4.4+/-0.2 mm). Regression analysis of the two series of data shows a R(2)=0.9984. IVUS measurements are an average 5% smaller than sonomicrometer measurements (3.6% up to 8.3%) and the difference is statistically significant ( p <0.05). The underestimation of IVUS measurements will affect the accuracy, and probably the long-term outcome, of endovascular procedures. PMID- 10828232 TI - Interobserver agreement in duplex scanning for vein grafts. AB - BACKGROUND: although the precision of duplex scanning is of utmost importance in vein-graft surveillance, it has not been properly assessed. This study aims to analyse interobserver agreement on duplex scanning. METHODS: a blinded comparative trial of 69 infrainguinal vein bypass reconstructions. Two consecutive duplex scans were performed by different examiners and duplex ultrasound machines on the same patient. The duplex examinations were also compared with angiography, when available, and clinical follow-up. RESULTS: interobserver agreement in Kappa statistics was 0.69, signifying "good" agreement between the examinations in detecting haemodynamically significant changes in the grafts. The sensitivity, specificity and accuracy figures compared with a combination of angiography and follow-up data for the two scans were 80%, 91%, 88% and 85%, 93%, 91%, respectively. The limits of agreement were, however, wide for Doppler-derived velocity characteristics. CONCLUSION: duplex scanning is an accurate and reproducible method for detecting haemodynamically significant changes in infrainguinal vein grafts. PMID- 10828233 TI - Lower-extremity revascularisation without preoperative contrast arteriography in 185 cases: lessons learned with duplex ultrasound arterial mapping. AB - PURPOSE: we have previously reported our experience with lower-extremity duplex ultrasound arterial mapping (DUAM) compared to contrast arteriography (CA) to predict lower-extremity bypass sites. The present study evaluates arterial revascularisation procedures for chronic limb ischaemia based on DUAM. MATERIALS AND METHODS: from January 1998 to July 1999, 195 patients (128 men, 67 women) underwent 211 lower-extremity revascularisation procedures based on DUAM. Indications for surgery were tissue loss, severe claudication, rest pain and popliteal aneurysm in 57%, 25%, 14% and 4% of the limbs, respectively. The mean age was 72+/-12 years and risk factors such as diabetes, hypertension, tobacco use, coronary artery and end-stage renal disease were present in 53%, 58%, 53%, 50% and 12% of the patients, respectively. Previous revascularisation procedures had been performed in 23% of the limbs. Preoperative evaluation consisted of DUAM alone (185) or of a combination of DUAM and CA (29 limbs). CA was deemed necessary due to a combination of technical difficulties that jeopardized adequate sonographic imaging and presence of disadvantaged run-off for medico legal reasons. DUAM consisted of direct imaging of all major arteries from the distal aorta to the pedal circulation. Optimal inflow and outflow bypass anastomotic sites were selected according to a diagram based on DUAM. Adequacy of the inflow was additionally assessed by common-femoral-artery waveform and confirmed by intraoperative pressure measurements. Post-bypass CA was obtained to verify patency of the run-off. RESULTS: DUAM procedure time averaged 75+/-26 min. For patients who underwent only DUAM, the distal anastomosis was to the popliteal artery in 91 cases and to tibial or pedal arteries in 58 cases. Distal anastomosis was proximal to a significant lesion in two cases that required jump grafts. Cumulative patency rates at 1 and 3 months for popliteal bypasses were 96% and 90%, and for infrapopliteal bypasses 90% and 83%, respectively. Inflow procedures to the femoral artery, patch and balloon angioplasties accounted for the remaining 40 cases. Four primary amputations were performed after CA confirmed DUAM findings. CONCLUSIONS: contrary to general belief, these data show that high-quality arterial ultrasonography represents a safe alternative to preoperative CA, even for infrapopliteal bypasses. This non-invasive approach may be especially useful for patients with contrast allergy or impaired renal function. PMID- 10828234 TI - Gadolinium-enhanced magnetic resonance angiography, digital subtraction angiography and duplex of the iliac arteries compared with intra-arterial pressure gradient measurements. AB - PURPOSE: to compare gadolinium-enhanced magnetic resonance angiography (Gd-MRA), digital subtraction angiography (DSA) and duplex of the iliac arteries with intra arterial pressure gradient measurement as the reference method. MATERIALS AND METHODS: Gd-MRA, DSA and duplex examinations of the iliac arteries were performed in 30 patients (60 arteries) with lower-limb arterial occlusive disease. In 29 arteries, pressure measurements were made (n=25) or the artery was found to be occluded on catheterisation (n=4). An aortofemoral peak systolic pressure gradient of 20 mmHg or more was regarded as haemodynamically significant. Stenoses with a diameter reduction of 50% or more on MRA or DSA, or an increase in peak systolic velocity greater than 150% (duplex) were considered significant. MRA examinations were evaluated by means of maximum intensity projections (MRA MIP) and using source images and curved multiplanar reconstruction (MRA-MPR). RESULTS: the sensitivity (specificity) for a significant iliac artery stenosis were 81% (75%) for MRA-MIP, 76% (75%) for MRA-MPR, 86% (88%) for DSA, and 72% (88%) for duplex. CONCLUSION: with intra-arterial pressure measurements as the reference method, similar results were achieved with Gd-MRA, DSA and duplex concerning the detection of haemodynamically significant iliac artery stenoses. The use of source images and multiplanar reconstructions resulted in higher accuracy for the detection of occlusions. PMID- 10828235 TI - Feasibility of using dynamic contrast-enhanced magnetic resonance angiography as the sole imaging modality prior to endovascular repair of abdominal aortic aneurysms. AB - OBJECTIVES: to establish the feasibility of using magnetic resonance imaging (MRI) with dynamic contrast-enhanced (DCE) MRA as the sole imaging modality in the assessment of patients prior to endovascular repair of abdominal aortic aneurysms (AAAs). DESIGN: DCE MRA with MRI and helical computed tomography (CT) examinations were performed in patients being assessed for suitability for an endovascular approach to repair of their AAA. Management outcomes determined by the two techniques were compared. MATERIALS: sixteen patients with AAA. METHODS: all subjects underwent DCE MRA/MRI and helical CTA. Criteria for suitability for endovascular repair were established. The management outcomes determined by the MRI findings were compared with those obtained by CTA. RESULTS: high-quality MRA/MRI and CT images were obtained in 16 patients. Six patients were considered suitable for an endovascular approach, one was considered borderline and nine were judged unsuitable. In all cases, the overall management determined by the two methods concurred. CONCLUSIONS: comparison of the two imaging modalities resulted in agreement as to suitability for an endovascular approach. We therefore conclude that in our group the use of MRI and DCE-MRA proved effective as a sole imaging modality for the assessment of these patients. PMID- 10828236 TI - Role of duplex scan in endoleak detection after endoluminal abdominal aortic aneurysm repair. AB - OBJECTIVE: to validate the role of duplex scan in endoleak detection in postoperative surveillance of endoluminal abdominal aneurysm repair (EAAR). PATIENTS AND METHODS: between April 1997 and March 1999, 103 patients were eligible for duplex and computed tomography (CT) scan after EAAR. Mean follow-up was 8 months (range 1-24 months). The study protocol comprised concurrent examination with colour-duplex and CT scan at 1, 6, and 12 months after EAAR, for a total of 198 concurrent examinations. All duplex scan examinations were performed by two vascular surgeons with the same machine (ATL HDI 3000). Interobserver agreement in endoleak detection (kappa=1) and in type of endoleak (kappa=0.7) was evaluated in 50 random duplex examinations. Endoleak detection was examined comparatively in duplex and CT scan, the latter being the gold standard. Sensitivity and specificity tests together with negative- and positive predictive values (NPV and PPV) were calculated. RESULTS: duplex scan was not feasible in one patient. On CT scan the endoleak rate was 4% at one month, 3% at 6 months, and 4% at one year. Overall, CT scan detected 12 endoleaks. With respect to endoleak detection, duplex scan revealed a great ability in ruling out false-negative results (sensitivity 91.7%, NPV 99.4%), but overestimated the presence of endoleak (specificity 98.4%, PPV 78. 6%). Regarding type of endoleak, the ability of duplex scan to identify the source of endoleak was low (sensitivity 66.7%). CONCLUSIONS: duplex scan, if validated, appears to be a reliable means for excluding the presence of endoleak after EAAR. PMID- 10828237 TI - The inflammatory response following treatment of abdominal aortic aneurysms: a comparison between open surgery and endovascular repair. AB - OBJECTIVES: to compare the inflammatory response following endovascular and conventional AAA repair. DESIGN: prospective study. PATIENTS AND METHODS: ten patients were selected for open surgery (OPEN) and ten for endovascular (ENDO) AAA repair. Leukocytes, platelets, myeloperoxidase, lactoferrin, beta thromboglobulin, C-reactive protein (CRP), interleukin 6 (IL-6), tumour necrosis factor alpha (TNF-alpha) and complement activation products were measured before, during and after surgery. RESULTS: in the OPEN group the median hospital stay was longer (6 vs. 12 days, p=0.001) and more patients required transfusion (p=0.02). IL-6 and CRP increased postoperatively, most in OPEN (p<0.01). Platelet counts decreased after the first angiography in ENDO (p<0.01) and before aortic cross clamping in OPEN (p<0.05). The decrease was larger in OPEN (p=0.02). Leukocyte counts decreased after the first angiography in ENDO, and thereafter increased (p=0.001). An equivalent increase was observed in OPEN after declamping (p=0.001). Leukocyte and platelet degranulation products increased after the first angiography in ENDO and after declamping in OPEN. Changes in complement activation products were small. TNF-alpha did not change significantly. CONCLUSION: endovascular AAA repair caused significant leukocyte and platelet activation. Based on the timing of activation this could be caused by radiographic contrast media. PMID- 10828238 TI - Arterial flow induces changes in saphenous vein endothelium proteins transduced by cation channels. AB - OBJECTIVES: expression of leukocyte adhesins and proteins controlling thrombosis is likely to be an important determinant of graft patency early following vein bypass. We have previously demonstrated rapid increase in endothelial ICAM-1 and nitric oxide synthase (eNOS) concentrations in human saphenous vein exposed to arterial flow. The aim of this study was to investigate whether ion-channel blocking drugs could alter these flow-induced changes. METHODS: human saphenous vein segments, freshly excised from patients, were placed in a validated in vitro circuit using flow conditions shown to simulate arterial or venous circulations for 90 min, in the presence or absence of ion-channel blockers. The concentrations of ICAM-1, VCAM-1, eNOS and tissue factor (TF) were assessed by quantitative immunohistochemistry in vein exposed to flow and compared with that in freshly excised vein from the same patient. The endothelial protein concentration was calculated as the mean area of staining as percentage of that for the control protein CD31, using computer-aided image analysis. RESULTS: after arterial flow conditions the area ratio of ICAM-1 increased from 21.4+/-1.4 to 44.6+/-2.0%, of eNOS increased from 50.0+/-5.6 to 70.1+/-5.0%, of VCAM-1 decreased from 16.6+/-3.4 to 3.6+/-1.0%, whereas TF staining area ratio was unchanged. Inclusion of the non-selective K(+)channel blocker, tetraethylammonium, in the arterial perfusion solution abolished all these arterial flow-induced changes. Inclusion of the K(+)ATP channel blocker, glibenclamide, selectively abolished the arterial flow-induced changes in ICAM-1 and VCAM-1. Inclusion of the calcium channel blocker, nifedipine, abolished the arterial flow-induced changes in eNOS and VCAM-1 but increased the TF staining area ratio from 3.0+/-0.4 to 8.5+/-0.7%, p=0.01. Inclusion of the stretch activated cation-channel blocker, gadolinium, enhanced the arterial flow-induced increase in eNOS, but prevented the arterial flow-induced increase in ICAM-1. CONCLUSIONS: perfusion of veins under arterial flow conditions with gadolinium was associated with low endothelial concentrations of ICAM-1, VCAM-1 and TF, but high levels of eNOS. Such a concentration of endothelial proteins may be advantageous in newly implanted vein grafts. In contrast, nifedipine could have adverse effects by promoting increase in TF concentration. PMID- 10828239 TI - Catastrophic arterial thromboembolism associated with factor V Leiden. PMID- 10828240 TI - A psoas ganglion causing obstruction of the iliofemoral arteries. PMID- 10828241 TI - AV fistulae. PMID- 10828242 TI - Localization and molecular determinants of the Hanatoxin receptors on the voltage sensing domains of a K(+) channel. AB - Hanatoxin inhibits voltage-gated K(+) channels by modifying the energetics of activation. We studied the molecular determinants and physical location of the Hanatoxin receptors on the drk1 voltage-gated K(+) channel. First, we made multiple substitutions at three previously identified positions in the COOH terminus of S3 to examine whether these residues interact intimately with the toxin. We also examined a region encompassing S1-S3 using alanine-scanning mutagenesis to identify additional determinants of the toxin receptors. Finally, guided by the structure of the KcsA K(+) channel, we explored whether the toxin interacts with the peripheral extracellular surface of the pore domain in the drk1 K(+) channel. Our results argue for an intimate interaction between the toxin and the COOH terminus of S3 and suggest that the Hanatoxin receptors are confined within the voltage-sensing domains of the channel, at least 20-25 A away from the central pore axis. PMID- 10828243 TI - Interactions of cyclic nucleotide-gated channel subunits and protein tyrosine kinase probed with genistein. AB - The cGMP sensitivity of cyclic nucleotide-gated (CNG) channels can be modulated by changes in phosphorylation catalyzed by protein tyrosine kinases (PTKs) and protein tyrosine phosphatases. Previously, we used genistein, a PTK inhibitor, to probe the interaction between PTKs and homomeric channels comprised of alpha subunits (RETalpha) of rod photoreceptor CNG channels expressed in Xenopus oocytes. We showed that in addition to inhibiting phosphorylation, genistein triggers a noncatalytic interaction between PTKs and homomeric RETalpha channels that allosterically inhibits channel gating. Here, we show that native CNG channels from rods, cones, and olfactory receptor neurons also exhibit noncatalytic inhibition induced by genistein, suggesting that in each of these sensory cells, CNG channels are part of a regulatory complex that contains PTKs. Native CNG channels are heteromers, containing beta as well as alpha subunits. To determine the contributions of alpha and beta subunits to genistein inhibition, we compared the effect of genistein on native, homomeric (RETalpha and OLFalpha), and heteromeric (RETalpha+beta, OLFalpha+beta, and OLFalpha+RETbeta) CNG channels. We found that genistein only inhibits channels that contain either the RETalpha or the OLFbeta subunits. This finding, along with other observations about the maximal effect of genistein and the Hill coefficient of genistein inhibition, suggests that the RETalpha and OLFbeta subunits contain binding sites for the PTK, whereas RETbeta and OLFalpha subunits do not. PMID- 10828244 TI - Dominant-negative mutants identify a role for GIRK channels in D3 dopamine receptor-mediated regulation of spontaneous secretory activity. AB - The human D3 dopamine receptor can activate G-protein-coupled inward rectifier potassium channels (GIRKs), inhibit P/Q-type calcium channels, and inhibit spontaneous secretory activity in AtT-20 neuroendocrine cells (Kuzhikandathil, E.V., W. Yu, and G.S. Oxford. 1998. Mol. Cell. Neurosci. 12:390-402; Kuzhikandathil, E.V., and G.S. Oxford. 1999. J. Neurosci. 19:1698-1707). In this study, we evaluate the role of GIRKs in the D3 receptor-mediated inhibition of secretory activity in AtT-20 cells. The absence of selective blockers for GIRKs has precluded a direct test of the hypothesis that they play an important role in inhibiting secretory activity. However, the tetrameric structure of these channels provides a means of disrupting endogenous GIRK function using a dominant negative approach. To develop a dominant-negative GIRK mutant, the K(+) selectivity amino acid sequence -GYG- in the putative pore domain of the human GIRK2 channels was mutated to -AAA-, -GLG-, or -GFG-. While the mutation of -GYG- to -GFG- did not affect channel function, both the -AAA- and -GLG- GIRK2 mutants were nonfunctional. This suggests that the aromatic ring of the tyrosine residue rather than its hydroxyl group is involved in maintaining the pore architecture of human GIRK2 channels. When expressed in AtT-20 cells, the nonfunctional AAA GIRK2 and GLG-GIRK2 acted as effective dominant-negative mutants and significantly attenuated endogenous GIRK currents. Furthermore, these dominant negative mutants interfered with the D3 receptor-mediated inhibition of secretion in AtT-20 cells, suggesting they are centrally involved in the signaling pathway of this secretory response. These results indicate that dominant-negative GIRK mutants are effective molecular tools to examine the role of GIRK channels in vivo. PMID- 10828245 TI - Role of domain 4 in sodium channel slow inactivation. AB - Depolarization of sodium channels initiates at least three gating pathways: activation, fast inactivation, and slow inactivation. Little is known about the voltage sensors for slow inactivation, a process believed to be separate from fast inactivation. Covalent modification of a cysteine substituted for the third arginine (R1454) in the S4 segment of the fourth domain (R3C) with negatively charged methanethiosulfonate-ethylsulfonate (MTSES) or with positively charged methanethiosulfonate-ethyltrimethylammonium (MTSET) produces a marked slowing of the rate of fast inactivation. However, only MTSES modification produces substantial effects on the kinetics of slow inactivation. Rapid trains of depolarizations (2-20 Hz) cause a reduction of the peak current of mutant channels modified by MTSES, an effect not observed for wild-type or unmodified R3C channels, or for mutant channels modified by MTSET. The data suggest that MTSES modification of R3C enhances entry into a slow-inactivated state, and also that the effects on slow inactivation are independent of alterations of either activation or fast inactivation. This effect of MTSES is observed only for cysteine mutants within the middle of this S4 segment, and the data support a helical secondary structure of S4 in this region. Mutation of R1454 to the negatively charged residues aspartate or glutamate cannot reproduce the effects of MTSES modification, indicating that charge alone cannot account for these results. A long-chained derivative of MTSES has similar effects as MTSES, and can produce these effects on a residue that does not show use-dependent current reduction after modification by MTSES, suggesting that the sulfonate moiety can reach a critical site affecting slow inactivation. The effects of MTSES on R3C are partially counteracted by a point mutation (W408A) that inhibits slow inactivation. Our data suggest that a region near the midpoint of the S4 segment of domain 4 plays an important role in slow inactivation. PMID- 10828246 TI - Functional coupling of the beta(1) subunit to the large conductance Ca(2+) activated K(+) channel in the absence of Ca(2+). Increased Ca(2+) sensitivity from a Ca(2+)-independent mechanism. AB - Coexpression of the beta(1) subunit with the alpha subunit (mSlo) of BK channels increases the apparent Ca(2+) sensitivity of the channel. This study investigates whether the mechanism underlying the increased Ca(2+) sensitivity requires Ca(2+), by comparing the gating in 0 Ca(2+)(i) of BK channels composed of alpha subunits to those composed of alpha+beta(1) subunits. The beta(1) subunit increased burst duration approximately 20-fold and the duration of gaps between bursts approximately 3-fold, giving an approximately 10-fold increase in open probability (P(o)) in 0 Ca(2+)(i). The effect of the beta(1) subunit on increasing burst duration was little changed over a wide range of P(o) achieved by varying either Ca(2+)(i) or depolarization. The effect of the beta(1) subunit on increasing the durations of the gaps between bursts in 0 Ca(2+)(i) was preserved over a range of voltage, but was switched off as Ca(2+)(i) was increased into the activation range. The Ca(2+)-independent, beta(1) subunit induced increase in burst duration accounted for 80% of the leftward shift in the P(o) vs. Ca(2+)(i) curve that reflects the increased Ca(2+) sensitivity induced by the beta(1) subunit. The Ca(2+)-dependent effect of the beta(1) subunit on the gaps between bursts accounted for the remaining 20% of the leftward shift. Our observation that the major effects of the beta(1) subunit are independent of Ca(2+)(i) suggests that the beta(1) subunit mainly alters the energy barriers of Ca(2+)-independent transitions. The changes in gating induced by the beta(1) subunit differ from those induced by depolarization, as increasing P(o) by depolarization or by the beta(1) subunit gave different gating kinetics. The complex gating kinetics for both alpha and alpha+beta(1) channels in 0 Ca(2+)(i) arise from transitions among two to three open and three to five closed states and are inconsistent with Monod-Wyman-Changeux type models, which predict gating among only one open and one closed state in 0 Ca(2+)(i). PMID- 10828247 TI - Timing of Ca(2+) release from intracellular stores and the electrical response of Limulus ventral photoreceptors to dim flashes. AB - Light-induced release of Ca(2+) from stores in Limulus ventral photoreceptors was studied using confocal fluorescence microscopy and the Ca(2+) indicator dyes, Oregon green-5N and fluo-4. Fluorescence was collected from a spot within 4 microm of the microvillar membrane. A dual-flash protocol was used to reconstruct transient elevations of intracellular free calcium ion concentration (Ca(i)) after flashes delivering between 10 and 5 x 10(5) effective photons. Peak Ca(i) increased with flash intensity to 138 +/- 76 microM after flashes delivering approximately 10(4) effective photons, while the latent period of the elevation of Ca(i) fell from approximately 140 to 21 ms. The onset of the light-induced elevation of Ca(i) was always highly correlated with that of the receptor potential. The time for Ca(i) to exceed 2 microM was approximately equal to that for the receptor potential to exceed 8 mV (mean difference; 2.2 +/- 6.4 ms). Ca(i) was also measured during steps of light delivering approximately 10(5) effective photons/s to photoreceptors that had been bleached with hydroxylamine so as to reduce their quantum efficiency. Elevations of Ca(i) were detected at the earliest times of the electrical response to the steps of light, when a significant receptor potential had yet to develop. Successive responses exhibited stochastic variation in their latency of up to 20 ms, but the elevation of Ca(i) and the receptor potential still rose at approximately the same time, indicating a shared process generating the latent period. Light-induced elevations of Ca(i) resulted from Ca(2+) release from intracellular stores, being abolished by cyclopiazonic acid (CPA), an inhibitor of endoplasmic reticulum Ca(2+) pumps, but not by removal of extracellular Ca(2+) ions. CPA also greatly diminished and slowed the receptor potential elicited by dim flashes. The results demonstrate a rapid release of Ca(2+) ions that appears necessary for a highly amplified electrical response to dim flashes. PMID- 10828248 TI - Dynamic control of deactivation gating by a soluble amino-terminal domain in HERG K(+) channels. AB - K(+) channels encoded by the human ether-a-go-go-related gene (HERG) are distinguished from most other voltage-gated K(+) channels by an unusually slow deactivation process that enables cardiac I(Kr), the corresponding current in ventricular cells, to contribute to the repolarization of the action potential. When the first 16 amino acids are deleted from the amino terminus of HERG, the deactivation rate is much faster (Wang, J., M.C. Trudeau, A.M. Zappia, and G.A. Robertson. 1998. J. Gen. Physiol. 112:637-647). In this study, we determined whether the first 16 amino acids comprise a functional domain capable of slowing deactivation. We also tested whether this "deactivation subdomain" slows deactivation directly by affecting channel open times or indirectly by a blocking mechanism. Using inside-out macropatches excised from Xenopus oocytes, we found that a peptide corresponding to the first 16 amino acids of HERG is sufficient to reconstitute slow deactivation to channels lacking the amino terminus. The peptide acts as a soluble domain in a rapid and readily reversible manner, reflecting a more dynamic regulation of deactivation than the slow modification observed in a previous study with a larger amino-terminal peptide fragment (Morais Cabral, J.H., A. Lee, S.L. Cohen, B.T. Chait, M. Li, and R. Mackinnon. 1998. Cell. 95:649-655). The slowing of deactivation by the peptide occurs in a dose-dependent manner, with a Hill coefficient that implies the cooperative action of at least three peptides per channel. Unlike internal TEA, which slows deactivation indirectly by blocking the channels, the peptide does not reduce current amplitude. Nor does the amino terminus interfere with the blocking effect of TEA, indicating that the amino terminus binding site is spatially distinct from the TEA binding site. Analysis of the single channel activity in cell attached patches shows that the amino terminus significantly increases channel mean open time with no alteration of the mean closed time or the addition of nonconducting states expected from a pore block mechanism. We propose that the four amino-terminal deactivation subdomains of the tetrameric channel interact with binding sites uncovered by channel opening to specifically stabilize the open state and thus slow channel closing. PMID- 10828249 TI - Electrogenic Na(+)/Ca(2+) exchange. A novel amplification step in squid olfactory transduction. AB - Olfactory receptor neurons (ORNs) from the squid, Lolliguncula brevis, respond to the odors l-glutamate or dopamine with increases in internal Ca(2+) concentrations ([Ca(2+)](i)). To directly asses the effects of increasing [Ca(2+)](i) in perforated-patched squid ORNs, we applied 10 mM caffeine to release Ca(2+) from internal stores. We observed an inward current response to caffeine. Monovalent cation replacement of Na(+) from the external bath solution completely and selectively inhibited the caffeine-induced response, and ruled out the possibility of a Ca(2+)-dependent nonselective cation current. The strict dependence on internal Ca(2+) and external Na(+) indicated that the inward current was due to an electrogenic Na(+)/Ca(2+) exchanger. Block of the caffeine induced current by an inhibitor of Na(+)/Ca(2+) exchange (50-100 microM 2',4' dichlorobenzamil) and reversibility of the exchanger current, further confirmed its presence. We tested whether Na(+)/Ca(2+) exchange contributed to odor responses by applying the aquatic odor l-glutamate in the presence and absence of 2', 4'-dichlorobenzamil. We found that electrogenic Na(+)/Ca(2+) exchange was responsible for approximately 26% of the total current associated with glutamate induced odor responses. Although Na(+)/Ca(2+) exchangers are known to be present in ORNs from numerous species, this is the first work to demonstrate amplifying contributions of the exchanger current to odor transduction. PMID- 10828250 TI - Interaction between quaternary ammonium ions in the pore of potassium channels. Evidence against an electrostatic repulsion mechanism. AB - We have examined the interaction between internal and external ions in the pore of potassium channels. We found that external tetraethylammonium was able to antagonize block of Shaker channels by internal TEA when the external and internal solutions contained K(+) ions. This antagonism was absent in solutions with Rb(+) as the only permeant ion. An externally applied trivalent TEA analogue, gallamine, was less effective than the monovalent TEA in inhibiting block by internal TEA. In addition, block by external TEA was little affected by changes in the concentration of internal K(+) ions, but was increased by the presence of internal Na(+) ions in the pore. These results demonstrate that external and internal TEA ions, likely located at opposite ends of the pore selectivity filter, do not experience a mutual electrostatic repulsion. We found that these results can be simulated by a simple 4-barrier-3-site permeation model in which ions compete for available binding sites without long-range electrostatic interactions. PMID- 10828251 TI - Mechanism of cGMP-gated channel block by intracellular polyamines. AB - Polyamines block the retinal cyclic nucleotide-gated channel from both the intracellular and extracellular sides. The voltage-dependent mechanism by which intracellular polyamines inhibit the channel current is complex: as membrane voltage is increased in the presence of polyamines, current inhibition is not monotonic, but exhibits a pronounced damped undulation. To understand the blocking mechanism of intracellular polyamines, we systematically studied the endogenous polyamines as well as a series of derivatives. The complex channel blocking behavior of polyamines can be accounted for by a minimal model whereby a given polyamine species (e.g., spermine) causes multiple blocked channel states. Each blocked state represents a channel occupied by a polyamine molecule with characteristic affinity and probability of traversing the pore, and exhibits a characteristic dependence on membrane voltage and cGMP concentration. PMID- 10828252 TI - Mechanism of IRK1 channel block by intracellular polyamines. AB - Intracellular polyamines inhibit the strongly rectifying IRK1 potassium channel by a mechanism different from that of a typical ionic pore blocker such as tetraethylammonium. As in other K(+) channels, in the presence of intracellular TEA, the IRK1 channel current decreases with increasing membrane voltage and eventually approaches zero. However, in the presence of intracellular polyamines, the channel current varies with membrane voltage in a complex manner: when membrane voltage is increased, the current decreases in two phases separated by a hump. Furthermore, contrary to the expectation for a nonpermeant ionic pore blocker, a significant residual IRK1 current persists at very positive membrane voltages; the amplitude of the residual current decreases with increasing polyamine concentration. This complex blocking behavior of polyamines can be accounted for by a minimal model whereby intracellular polyamines inhibit the IRK1 channel by inducing two blocked channel states. In each of the blocked states, a polyamine is bound with characteristic affinity and probability of traversing the pore. The proposal that polyamines traverse the pore at finite rates is supported by the observation that philanthotoxin-343 (spermine with a bulky chemical group attached to one end) acts as a nonpermeant ionic blocker in the IRK1 channel. PMID- 10828253 TI - Microsomal enzyme induction. PMID- 10828254 TI - Why are some proteins allergens? AB - The ability of certain proteins to induce an allergic response in susceptible individuals is well established. Symptoms can range from mild erythema or rhinitis, to acute, and possibly fatal, anaphylactic shock. Because such allergic responses require complex interactions between the protein and the immune system, they are notoriously difficult to predict. Nevertheless, it is clear that some proteins are intrinsically more allergenic than others. The challenge for toxicologists is to identify those characteristics that confer on proteins the potential to induce allergic sensitization and allergic disease. Here, we first consider the potential contribution that individual epitopes may make to the allergenicity of a protein. These are the minimal peptide units within proteins that can be recognized by the immune system and are a fundamental requirement for all immune responses, including those resulting in allergic sensitization. It appears that allergens must necessarily contain B-cell epitopes to which immunoglobulin E (IgE) can bind, and T-cell epitopes capable of inducing a type 2 T-lymphocyte response. Nevertheless, it appears doubtful that the presence of appropriate epitopes alone is sufficient to endow a protein with allergenic potential. We therefore consider also the contribution that other features and characteristics of proteins may make to their overall allergenicity. In particular, we consider the effects that resistance to proteolysis, post translational glycosylation, and enzymatic activity may have. It appears that relative stability in simulated gastric fluid (SGF) sometimes correlates with allergenic activity. However, this is not universally true, and it is known that there are protein allergens, such as some of those associated with oral allergy syndrome, that are unstable. Nevertheless, if stability in SGF is associated with the intrinsic allergenicity of many proteins irrespective of the route of exposure, then this may reflect some more fundamental property of proteins, and possibly their stability in other biologic matrices and/or to intracellular proteases. Post-translational modification appears generally to enhance allergenicity, perhaps by increasing uptake and detection of the protein by the immune system. Some enzymatic activities also enhance allergenicity through what appear to be several different mechanisms, including nonspecific activation of cells participating in the immunologic response. Overall, it appears likely that many factors can contribute to the overall allergenicity of any given protein. Some, such as the presence of epitopes with allergenic potential, may be essential. Others, such as the glycosylation status, resistance to proteolysis, and enzymatic activity, may play a subsidiary but nevertheless critically important role. By better defining the limits within which these factors operate, we can hope to gain a better understanding of the fundamental origins of protein allergenicity, and therefore be in a position to identify and characterize the hazards and risks of allergic disease associated with novel proteins. PMID- 10828255 TI - Assessment of skin absorption and penetration of JP-8 jet fuel and its components. AB - Dermal penetration and absorption of jet fuels in general, and JP-8 in particular, is not well understood, even though government and industry, worldwide, use over 4.5 billion gallons of JP-8 per year. Exposures to JP-8 can occur from vapor, liquid, or aerosol. Inhalation and dermal exposure are the most prevalent routes. JP-8 may cause irritation during repeated or prolonged exposures, but it is unknown whether systemic toxicity can occur from dermal penetration of fuels. The purpose of this investigation was to measure the penetration and absorption of JP-8 and its major constituents with rat skin, so that the potential for effects with human exposures can be assessed. We used static diffusion cells to measure both the flux of JP-8 and components across the skin and the kinetics of absorption into the skin. Total flux of the hydrocarbon components was 20.3 micrograms/cm(2)/h. Thirteen individual components of JP-8 penetrated into the receptor solution. The fluxes ranged from a high of 51.5 micrograms/cm(2)/h (an additive, diethylene glycol monomethyl ether) to a low of 0.334 micrograms/cm(2)/h (tridecane). Aromatic components penetrated most rapidly. Six components (all aliphatic) were identified in the skin. Concentrations absorbed into the skin at 3.5 h ranged from 0.055 micrograms per gram skin (tetradecane) to 0.266 micrograms per gram skin (undecane). These results suggest: (1) that JP-8 penetration will not cause systemic toxicity because of low fluxes of all the components; and (2) the absorption of aliphatic components into the skin may be a cause of skin irritation. PMID- 10828256 TI - Application of a physiologically based pharmacokinetic model to estimate the bioavailability of ethanol in male rats: distinction between gastric and hepatic pathways of metabolic clearance. AB - A portion of ingested ethanol does not reach the systemic circulation in both rats and humans as indicated by higher blood ethanol concentrations following an intravenous administration compared to an equivalent oral administration. The mechanism for this decrease in the oral bioavailability is not yet completely understood. Metabolism by gastric or hepatic alcohol dehydrogenase (ADH), or both, has been implicated. However, the extent to which each pathway of elimination contributes to the first-pass clearance is not known. The purpose of this study was to utilize a physiologically based pharmacokinetic (PBPK) model for ethanol to estimate the relative contributions of hepatic and gastric metabolic clearance to the oral bioavailability of ethanol in male rats. In the current model, calculations of hepatic-first pass metabolic clearance accounted for the competition for metabolism between incoming ethanol from the GI tract and recirculating ethanol. This differs from previous methods that quantified the effect of ethanol entering the liver from the GI tract on the overall rate of metabolism of ethanol by the liver. These models did not specifically describe the effect of recirculating ethanol on the first-pass metabolism of ethanol, and vice versa. The dependence of bioavailability on dose and absorption rate was also investigated. The use of a PBPK model for ethanol in rats allows a more detailed examination of physiological and biochemical factors affecting the bioavailability of ethanol than has previously been possible. The analysis indicates that both gastric and hepatic first-pass metabolism of ethanol contribute to ethanol bioavailability in male rats. PMID- 10828257 TI - Inhibition of cytochrome P450 2E1 decreases, but does not eliminate, genotoxicity mediated by 1,3-butadiene. AB - 1,3-Butadiene (BD), a rodent carcinogen, is metabolized to mutagenic and DNA reactive epoxides. In vitro data suggest that this oxidation is mediated by cytochrome P450 2E1 (CYP2E1). In this study, we tested the hypothesis that oxidation of BD by CYP2E1 is required for genotoxicity to occur. Inhalation exposures were conducted with B6C3F1 mice using a closed-chamber technique, and the maximum rate of butadiene oxidation was estimated. The total amount of butadiene metabolized was then correlated with the frequency of micronuclei (MN). Three treatment groups were used: (1) mice with no pretreatment; (2) mice pretreated with 1,2-trans-dichloroethylene (DCE), a specific CYP2E1 inhibitor; and (3) mice pretreated with 1-aminobenzotriazole (ABT), an irreversible inhibitor of cytochromes P450. Mice in all 3 groups were exposed to an initial BD concentration of 1100 ppm, and the decline in concentration of BD in the inhalation chamber with time, due to uptake and metabolism of BD, was monitored using gas chromatography. A physiologically based pharmacokinetic model was used to analyze the gas uptake data, estimate V(max) for BD oxidation, and compute the total amount of BD metabolized. Model simulations of the gas uptake data predicted the maximum rate of BD oxidation would be reduced by 60% and 100% for the DCE- and ABT-pretreated groups, respectively. Bone marrow was harvested 24 h after the onset of the inhalation exposure and analyzed for frequency of micronuclei in polychromatic erythrocytes (MN-PCE). The frequency of MN-PCE per 1000 PCE in mice exposed to BD was 28.2 +/- 3.1, 19.8 +/- 2.5, and 12.3 +/- 1.9, for the mice with no pretreatment, DCE-pretreated mice and ABT-pretreated mice, respectively. Although inhibition of CYP2E1 decreased BD-mediated genotoxicity, it did not completely eliminate genotoxic effects. These data suggest that other P450 isoforms may contribute significantly to the metabolic activation of BD and resultant genotoxicity. PMID- 10828258 TI - Biotransformation and kinetics of excretion of tert-amyl-methyl ether in humans and rats after inhalation exposure. AB - tert-Amyl methyl ether (TAME) may be widely used as an additive to gasoline in the future. The presence of this ether in gasoline reduces the tail pipe emission of pollutants. Therefore, widespread human exposure to TAME may occur. To contribute to the characterization of potential adverse effects of TAME, its biotransformation was compared in humans and rats after inhalation exposure. Human volunteers (three males and three females) and rats (five males and five females) were exposed to 4 (3.8 +/- 0.2) and 40 (38.4 +/- 1.7) ppm TAME for 4 h in a dynamic exposure system. Urine samples were collected for 72 h in 6-h intervals and blood samples were taken at regular intervals for 48 h in humans. In urine, the TAME metabolites tert-amyl alcohol (t-amyl alcohol), 2-methyl-2, 3 butane diol, 2-hydroxy-2-methylbutyric acid, and 3-hydroxy-3-methylbutyric acid were quantified. TAME and t-amyl alcohol were determined in blood samples. After the end of the exposure period, blood concentrations of TAME were 4.4 +/- 1.7 microM in humans and 9.6 +/- 1.4 microM in rats after 40 ppm TAME, and 0.6 +/- 0.1 microM in humans and 1.4 +/- 0.8 microM in rats after 4 ppm. TAME was rapidly cleared from blood in both rats and humans. The blood concentrations of t-amyl alcohol were 9.2 +/- 1.8 microM in humans and 8.1 +/- 1.5 microM in rats after 40 ppm TAME, and 1.0 +/- 0.3 microM in humans and 1.8 +/- 0.2 microM in rats after 4 ppm TAME. t-Amyl alcohol was also rapidly cleared from blood. In urine of humans, 2-methyl-2,3-butane diol, 2-hydroxy-2-methylbutyric acid, and 3-hydroxy-3 methylbutyric acid were recovered as major excretory products in urine. In rats, 2-methyl-2,3-butane diol and its glucuronide were major TAME metabolites. t-Amyl alcohol and its glucuronide were minor TAME metabolites in both species. All metabolites of TAME excreted with urine in rats were rapidly eliminated, with elimination half-lives of less than 6 h. Metabolite excretion in humans was slower and elimination half-lives of the different metabolites were between 6 and 40 h in humans. The obtained data indicate differences in TAME biotransformation and excretion between rats and humans. In rats, TAME metabolites are rapidly excreted. In humans, metabolic pathways are different and metabolite excretion is slower. Recovery of TAME metabolites in urine was higher in humans as compared to rats, suggesting more intensive biotransformation of TAME in humans. PMID- 10828259 TI - Metabolism of 3-methylindole by porcine liver microsomes: responsible cytochrome P450 enzymes. AB - The role of different cytochrome P450 enzymes on the metabolism of 3-methylindole (3MI) was investigated using selective chemical inhibitors. Eight chemical inhibitors of P450 enzymes were screened for their inhibitory specificity towards 3MI metabolism in porcine microsomes: alpha-naphthoflavone (CYP1A1/2), 8 methoxypsoralen (CYP2A6), menthofuran (CYP2A6), diethyldithiocarbamate (CYP2A6), 4-methylpyrazole (CYP2E1), sulphaphenazole (CYP2C9), quinidine (CYP2D6), and troleandomycin (CYP3A4). The production of 3MI metabolites was only affected by the presence of inhibitors of CYP2A6 and CYP2E1 in the microsomal incubations. In a second experiment, a set of porcine microsomes (n = 30) was analyzed for CYP2A6 content by protein immunoblot analysis and for their coumarin 7-hydroxylation activity (CYP2A6 activity). Both CYP2A6 content and enzymatic activity were found to be highly and negatively correlated with 3MI fat content. The results of the present study indicate that the CYP2A6 porcine ortholog plays an important role in the metabolism of 3MI and that measurement of CYP2A6 levels and/or activity could be a useful marker for 3MI-induced boar taint. PMID- 10828260 TI - Statistical analysis of skin tumor data from Tg.AC mouse bioassays. AB - New strategies for identifying chemical carcinogens and assessing risk have been proposed based on the Tg.AC (zetaglobin promoted v-Ha-ras) transgenic mouse. Preliminary studies suggest that the Tg. AC mouse bioassay may be an effective means of quickly evaluating the carcinogenic potential of a test agent. The skin of the Tg.AC mouse is genetically initiated, and the induction of epidermal papillomas in response to dermal or oral exposure to a chemical agent acts as a reporter phenotype of the activity of the test chemical. In Tg.AC mouse bioassays, the test agent is typically applied topically for up to 26 weeks, and the number of papillomas in the treated area is counted weekly. Statistical analyses are complicated by within-animal and serial dependency in the papilloma counts, survival differences between animals, and missing data. In this paper, we describe a statistical model for the analysis of skin tumor data from a Tg.AC mouse bioassay. The model separates effects on papilloma latency and multiplicity and accommodates important features of the data, including variability in expression of the transgene and dependency in the tumor counts. Methods are described for carcinogenicity testing and risk assessment. We illustrate our approach using data from a study of the effect of 2,3,7, 8-tetrachlorodibenzo-p dioxin (TCDD) exposure on tumorigenesis. PMID- 10828261 TI - Choline supplementation inhibits diethanolamine-induced morphological transformation in syrian hamster embryo cells: evidence for a carcinogenic mechanism. AB - DEA, an amino alcohol, and its fatty acid condensates are widely used in commerce. DEA is hepatocarcinogenic in mice, but shows no evidence of mutagenicity or clastogenicity in a standard testing battery. However, it increased the number of morphologically transformed colonies in the Syrian hamster embryo (SHE) cell morphologic transformation assay. The goal of this work was to test the hypothesis that DEA treatment causes morphologic transformation by a mechanism involving altered cellular choline homeostasis. As a first step, the ability of DEA to disrupt the uptake and intracellular utilization of choline was characterized. SHE cells were cultured in medium containing DEA (500 microg/ml), and (33)P-phosphorus or (14)C-choline was used to label phospholipid pools. After 48 h, SHE cells were harvested, lipids were extracted, and radioactive phospholipids were quantified by autoradiography after thin layer chromatographic separation. In control cells, phosphatidylcholine (PC) was the major phospholipid, accounting for 43 +/- 1% of total phospholipid synthesis. However, with DEA treatment, PC was reduced to 14 +/- 2% of total radioactive phospholipids. DEA inhibited choline uptake into SHE cells at concentrations > or = 50 microg /ml, reaching a maximum 80% inhibition at 250-500 microg/ml. The concentration dependence of the inhibition of PC synthesis by DEA (0, 10, 50, 100, 250, and 500 microg/ml) was determined in SHE cells cultured over a 7-day period under the conditions of the transformation assay and in the presence or absence of excess choline (30 mM). DEA treatment decreased PC synthesis at concentrations > or = 100 microg/ml, reaching a maximum 60% reduction at 500 microg/ml. However, PC synthesis was unaffected when DEA-treated cells were cultured with excess choline. Under 7-day culture conditions, (14)C-DEA was incorporated into SHE lipids, and this perturbation was also inhibited by choline supplementation. Finally, DEA (10-500 microg/ml) transformed SHE cells in a concentration-dependent manner, whereas with choline supplementation, no morphologic transformation was observed. Thus, DEA disrupts intracellular choline homeostasis by inhibiting choline uptake and altering phospholipid synthesis. However, excess choline blocks these biochemical effects and inhibits cell transformation, suggesting a relationship between the two responses. Overall, the results provide a plausible mechanism to explain the morphologic transformation observed with DEA and suggest that the carcinogenic effects of DEA may be caused by intracellular choline deficiency. PMID- 10828262 TI - Effects of genistein exposure on sexually dimorphic behaviors in rats. AB - The phytoestrogen genistein, the principal isoflavone in soybeans, has adverse effects on animal reproduction. As adult physiology and behavior are sensitive to perturbation by developmental estrogens, exposure to genistein during development may produce behavioral alterations as well. Pregnant rats were fed soy-free diets containing 0, 25, 250, or 1250 ppm genistein (approximately 0, 2, 20, or 100 mg/kg/day) beginning on gestational day 7, and offspring continued on these diets through postnatal day (PND) 77. Male and female offspring were assessed for levels of sexually dimorphic behaviors: open field activity, play behavior, running wheel activity, and consumption of saccharin- and sodium chloride flavored solutions. Consumption of the salt solution was affected by genistein, with animals in the 1250-ppm group drinking significantly more than controls; consumption of plain water was unaffected. Genistein treatment also significantly affected play behavior; although no treated group was significantly different from controls, and the effect was not sexually dimorphic. Running wheel activity and saccharin solution consumption showed significant sex differences, but no effects of genistein treatment. Gestational duration, total and live pups per litter, and total and live litter sex ratios were not significantly affected by genistein. However, average weight per live pup at birth and offspring body weights from PND 42-77 were significantly decreased in the 1250-ppm group. Body weight and food intake for the dams were also significantly decreased in the 1250 ppm group. These results indicate that developmental genistein treatment, at levels that decrease maternal and offspring body weight, causes subtle alterations in some sexually dimorphic behaviors. PMID- 10828263 TI - Activity of benzo[a]pyrene and its hydroxylated metabolites in an estrogen receptor-alpha reporter gene assay. AB - A human breast cancer cell line, MCF-7, transiently transfected with a chimeric estrogen receptor (Gal4-HEG0) and a luciferase reporter plasmid (17m5-G-Luc), was used to investigate the estrogenic activity of benzo[a]pyrene (B[a]P), a prototypical polyaromatic hydrocarbon (PAH). B[a]P at concentrations > or = 1 microM produced responses comparable to that of 0.1 nM 17beta-estradiol (E2). The ER antagonist ICI 182,780 (ICI) completely inhibited the response to both E2 and B[a]P, indicating that the responses were ER-mediated. However, 2 microM alpha napthoflavone (alpha-NF), an Ah receptor antagonist and P450 inhibitor, also decreased the response to B[a]P but not to E2. Analysis of the profile of B[a]P metabolites in the transfected MCF-7 cultures indicated that alpha-NF inhibited the production of the 3- and 9-hydroxy (3-OH and 9-OH), as well as the 7, 8- and 9,10-dihydroxy (7,8-OH and 9,10-OH) B[a]P species. In the ER-alpha reporter assay, the 3-OH and 9-OH metabolites produced maximal responses comparable to E2, with EC50 values of 1.2 microM and 0.7 microM, respectively. The 9,10-OH metabolite exhibited minimal activity in the assay. These responses were inhibited by ICI for both the 3-OH and the 9-OH species; however, alpha-NF inhibited only the response to the 9-OH metabolite. The 7,8-OH metabolite did not exhibit significant estrogenic activity. Furthermore, 7,8-OH B[a]P displayed observable cytotoxicity at concentrations > or = 10(-7) M. This cytotoxic response was completely inhibited by alpha-NF, suggesting that 7,8-OH B[a]P was being further metabolized to one or more cytotoxic metabolites. PMID- 10828264 TI - Depletion of cellular protein thiols as an indicator of arylation in isolated trout hepatocytes exposed to 1,4-benzoquinone. AB - A method to measure protein thiols (PrSH), reduced and oxidized, was adapted to determine PrSH depletion in isolated rainbow trout hepatocytes exposed to arylating agent 1,4-benzoquinone (BQ). Toxicant analysis revealed rapid conversion of BQ to 1, 4-hydroquinone (HQ) upon addition to hepatocytes. Hepatocytes exposed to 200 microM BQ+HQ showed 80% decline in glutathione (GSH) (1 h), 30% loss of PrSH (6 h), and no loss of viability (24 h). Recoverable oxidized PrSH was detected only after 24 h (200 microM BQ+HQ). Exposure to 600 microM BQ+HQ caused rapid (10 min) loss of > 90% GSH and > 60% PrSH, with eventual cell death. Half of the PrSH depletion at 6 h observed in hepatocytes exposed to 600 microM BQ+HQ was recoverable by reduction with dithiothreitol. Following the loss of GSH in hepatocytes exposed to 600 microM BQ+HQ, cellular PrSH were susceptible to direct arylation and oxidation. Rainbow trout hepatocytes, which contained 10-fold less GSH than rat cells, had a GSH:PrSH ratio of 1:82 compared with rat ratios of 1:2 to 1:6. The methods reported are useful for further study and discrimination of reactive modes of action needed for prediction of aquatic organism susceptibility to these types of toxicants. PMID- 10828265 TI - Oral treatment with trimethoprim-sulfamethoxazole and zidovudine suppresses murine accessory cell-dependent immune responses. AB - Trimethoprim-sulfamethoxazole (TMP-SMX), commonly used for prophylaxis of Pneumocystis carinii pneumonia (PCP) in AIDS patients, often produces a high incidence of treatment-limiting reactions. We investigated the effect of oral administration of TMP-SMX alone or in combination with the antiretroviral drug zidovudine (ZDV) on hematopoiesis and cellular immunity in BALB/c mice. Daily treatment for 28 days with TMP-SMX (160:800 mg/kg) had no effect on hematopoiesis or the ex vivo proliferative response of splenic T lymphocytes to allogeneic tumor cells (EL-4) or to concanavalin A (ConA), or that of splenic B cells to lipopolysaccharide (LPS). ZDV at 240 mg/kg/day was not immunosuppressive but caused a mild macrocytic anemia. Combined treatment produced severe pancytopenia, a significant drop in splenic cellularity, and a 61% decrease in the percentage of splenic macrophages. The percentage of splenic CD3+ lymphocytes increased 150% in the TMP-SMX + ZDV group, but the ratios of T-cell subsets and the frequency of B cells remained unchanged. Combined drug treatment did not impair the proliferative response of B cells to LPS or that of T cells to EL-4 cells. In concert with the reduction in the percentage of macrophages, the proliferative response of T lymphocytes to ConA decreased significantly. Optimal ConA-induced T cell proliferation requires the participation of accessory cells (AC) (e.g., macrophages); EL-4 cells are able to function as AC. These data indicate that ZDV synergizes with TMP-SMX, causing severe hematotoxicity and suppressing AC dependent immune function, and suggest that this therapeutic regimen may contribute to the immune deterioration in AIDS patients. PMID- 10828266 TI - Immunological responses of mice following administration of natural rubber latex proteins by different routes of exposure. AB - Although the prevalence of IgE-mediated latex allergy has increased over the past decade, the circumstances which culminate in sensitization remain uncertain. The objective of these studies was to evaluate the role which sensitization route plays in the development of latex allergy using murine models representative of potential exposure routes by which health care workers (topical and respiratory) and spina bifida patients (subcutaneous) may be sensitized. BALB/c mice administered latex proteins by the subcutaneous, topical, intranasal, or intratracheal routes exhibited dose-responsive elevations in total IgE. In vitro splenocyte stimulation initially demonstrated specificity of the murine immune response to latex proteins. Subsequently, immunoblot analysis was used to compare latex-specific IgE production amongst sensitization routes. Immunoblots of IgE from subcutaneously sensitized mice demonstrated recognition of latex proteins with molecular weights near 14 kDa and 27 kDa. These protein sizes are consistent with the molecular weights of major latex allergens (Hev b 1 and Hev b 3), to which high percentages of spina bifida patients develop antibodies. Mice sensitized by intratracheal or topical administration exhibited combined IgE recognition of latex proteins near 14 kDa, 35 kDa, and 92 kDa. These molecular weights are similar to other latex allergens (Hev b 6, Hev b 2, and Hev b 4) commonly recognized by IgE of health care workers. Mice sensitized to latex proteins by topical, intranasal, or intratracheal exposures exhibited bronchoconstriction as evaluated by whole body plethysmography following respiratory challenge with latex proteins. Subcutaneously sensitized mice were unresponsive. These differences in latex-specific IgE immunoblot profiles and altered pulmonary function amongst the four different sensitization routes suggest that exposure routes leading to sensitization may play a role in determining the primary allergen(s), and the clinical manifestation of the allergic responses. PMID- 10828267 TI - Fingerprinting of cytochrome P450 and microsomal epoxide hydrolase gene expression in human blood cells. AB - To examine the character and variability of human cytochrome P450 (CYP) and microsomal epoxide hydrolase (mEH) gene expression in human blood cells, we used a highly sensitive, quantitative, competitive reverse transcriptase-coupled polymerase chain reaction (QC RT-PCR) assay to assess mRNA profiles for a battery of 8 genes, in peripheral lymphocytes isolated from 10 healthy donors. Of the genes profiled, in lymphocytes CYP2D6 was typically expressed at the highest levels (3.8 x 10(5) molecules/microg total RNA), with CYP2E1 and mEH also maintained at relatively high abundance (1.2 x 10(5) and 1.8 x 10(5) molecules/microg total RNA, respectively). CYP1A1 levels were approximately an order of magnitude lower (3.9 x 10(4) molecules/microg total RNA), followed by CYP2F1 and CYP3A levels that were near the detection limit of the assay. CYP1A2 and CYP2A6/7 mRNAs were not detected in any of the lymphocyte samples. Overall, relatively low levels of inter-individual variation (2- to 6-fold) existed among these endpoint parameters in the subjects tested. To test whether established human blood cell lines were suitable models to assess basal expression and chemical induction responsiveness of these genes, we determined that constitutive CYP and mEH mRNA profiles were essentially conserved across 4 established human blood cell lines, and highly analogous to the basal expression patterns identified in freshly isolated peripheral lymphocytes. mEH protein was detected in all of the cell lines using Western immunoblotting and chemiluminescent visualization, whereas CYP1A1, CYP2D6, CYP2E1 or CYP3A proteins were not detected in these analyses. When blood cell-derived cultures were exposed to the prototypical CYP1A and CYP3A inducers, i.e., beta-naphthoflavone (beta-NF), dexamethasone (DEX) or phenobarbital, generally little or no inductive response was manifested. Thus, the data obtained from this investigation indicate that, although human blood cell lines in general exhibit poor responsiveness to prototypical inducer exposures, the constitutive patterns of CYP and mEH expression in peripheral lymphocytes appear to exhibit relatively low levels of variation among individuals. In addition, these in vivo patterns of expression are well maintained in established cultured blood-cell lines. PMID- 10828268 TI - Evaluation of cytotoxicity, cell proliferation, and genotoxicity induced by p cresidine in hetero- and nullizygous transgenic p53 mice. AB - The heterozygous p53 knockout mouse is being used as a short-term alternative model for carcinogenicity screening of chemicals. In most cases, these mice develop tumors within 6 months of exposure to genotoxic carcinogens. The bladder and liver carcinogen, p-cresidine, is recommended as a positive control chemical for these assays. To evaluate early effects of p53 deficiency on bladder and liver histopathology and genotoxicity induced by p-cresidine, we treated 4-week old heterozygous and nullizygous p53 male mice with p-cresidine by gavage (100, 200, 400, and 800 mg/kg/day) 5 days/week for 7 weeks. Tissue sections were prepared for hematoxylin-eosin staining and immunohistochemistry for PCNA protein or 3'-OH DNA fragments to assess cell proliferation and apoptosis, respectively. Blood and bone marrow were examined for methemoglobin and micronuclei in polychromatic erythrocytes (MN-PCE), respectively. Individual cell necrosis of the bladder transitional epithelium was evident in both p53 heterozygous and nullizygous mice at all doses. In addition, diffuse hyperplasia of the bladder epithelium was observed at 400 and 800 mg/kg in both genotypes. In the liver, both genotypes exhibited similar increases in hepatocyte apoptosis (10-fold increase) and cell proliferation (20-fold increase) at 800 mg/kg/day. Methemoglobin levels were increased 6-fold in both genotypes at 800 mg/kg. Background MN-PCE rates were similar in both genotypes and there were no treatment-related increases. Also, no point mutations were observed in codon 12 of the c-Ha-ras gene from urinary bladder DNA from p-cresidine treated p53 mice. These results suggest that loss of p53 allele(s) in mice does not influence the early markers of carcinogenic activity induced by subchronic treatment with p cresidine. Increased tumor susceptibility associated with a reduction in p53 dosage may be dependent on neoplastic progression rather than initiation and promotional events elicited by p-cresidine. PMID- 10828269 TI - Acetaminophen inhibits NF-kappaB activation by interfering with the oxidant signal in murine Hepa 1-6 cells. AB - A toxic dose of acetaminophen (APAP) reduces the activity of NF-kappaB in mouse liver. NF-kappaB inactivation may be important for APAP toxicity, as this transcription factor can play a central role in maintaining hepatic viability. We recently reported that APAP likewise inhibits serum growth factor activation of NF-kappaB in a mouse hepatoma cell line (Hepa 1-6 cells). Here we present evidence that APAP's antioxidant activity may be involved in this NF-kappaB inhibition in Hepa 1-6 cells. Like the antioxidants N-acetylcysteine (NAC) and pyrrolidinedithiocarbamate (PDTC), APAP was found to suppress the H(2)O(2) induced oxidation of an intracellular reactive oxygen species probe (dihydrodichlorofluorescein) in Hepa 1-6 cells. Treatment of Hepa 1-6 cells with H(2)O(2) was sufficient for NF-kappaB activation and IkappaBalpha degradation, and APAP was able to block both of these events. The APAP inhibition of NF-kappaB activation by serum growth factors may also be due to APAP's antioxidant activity, as the antioxidants NAC and PDTC likewise inhibit this activation. The potential role of NF-kappaB and oxidant-based growth factor signal transduction in APAP toxicity is discussed. PMID- 10828270 TI - Lack of mutagenic activity of 1,6-hexamethylene diisocyanate. AB - 1,6-Hexamethylene diisocyanate (HDI) is an aliphatic diisocyanate used in the manufacture of higher molecular weight biuret and trimer polyisocyanate resins. These resins are commonly used in polyurethane paints, resulting in potential occupational, and to a lesser extent consumer exposures. Because some isocyanates have been reported to be mutagenic, HDI was tested in the bacterial reverse mutation assay (Ames test), CHO/HGPRT gene mutation assay, and in the mouse micronucleus test, using vapor-phase exposures. Although indicators of toxicity were observed in each test, HDI did not induce mutagenic or clastogenic effects in any of the three assays. PMID- 10828271 TI - Intranasal instillation of aflatoxin B(1) in rats: bioactivation in the nasal mucosa and neuronal transport to the olfactory bulb. AB - Aflatoxin B(1) (AFB(1)) may be present in moldy dust. Inhalation of contaminated dust particles may result in high local exposure of the nasal mucosa. The present study was designed to assess bioactivation and toxicity of AFB(1) in the nasal mucosa after intranasal administration of the mycotoxin in rats and also to examine if translocation of the mycotoxin occurs from the nasal mucosa to the brain along olfactory neurons. Female Sprague-Dawley rats were given (3)H-AFB(1) (0.2, 1 or 20 microg) intranasally and were sacrificed at various intervals (1 h to 20 d). Tissues were examined autoradiographically or histopathologically. Quantitative data were obtained by beta-spectrometry in rats given (3)H-AFB(1) intranasally or orally (for comparison). The data indicated that intranasal administration of AFB(1) resulted in formation of tissue-bound metabolites in sustentacular cells, in some cells of Bowman's glands, and in a population of neuronal cells in the olfactory mucosa, whereas in the respiratory nasal mucosa, there was selective bioactivation of AFB(1) in mucous cells. Intranasal instillation of 20 microg AFB(1) resulted in disorganized undulating olfactory epithelium, with injured neuronal and sustentacular cells. In the respiratory epithelium, there was selective destruction of mucous cells. beta-Spectrometry and autoradiography with tape-sections of the head of rats given (3)H-AFB(1) intranasally indicated transport of AFB(1) and/or AFB(1) metabolites along the axons of the primary olfactory neurons to their terminations in the glomeruli of the olfactory bulb. The data indicate that the materials transported in the olfactory nerves represent AFB(1) and/or some of its nonreactive metabolites. It is concluded that application of AFB(1) on the nasal mucosa in rats results in high local bioactivation of the mycotoxin in this tissue and translocation of AFB(1) and/or its metabolites to the olfactory bulb. PMID- 10828272 TI - Manganese taken up into the CNS via the olfactory pathway in rats affects astrocytes. AB - Manganese (Mn), administered intranasally in rats, is effectively taken up in the CNS via the olfactory system. In the present study, Mn (as MnCl(2)) dissolved in physiological saline, was instilled intranasally in rats at doses of 0 (control), 10, 250, or 1000 microg. At the start of the experiment each rat received an intranasal instillation. Some rats were killed after one week without further treatment (the 1-w group), whereas the remaining rats received further instillations after one and two weeks and were killed after an additional week (the 3-w group). The brains were removed and either used for ELISA-determination of the astrocytic proteins glial fibrillary acidic protein (GFAP) and S-100b or histochemical staining of GFAP and S-100b, microglia (using an antibody against the iba1-protein) and the neuronal marker Fluoro-Jade. There were no indications that the Mn induced neuronal damage. On the other hand, the ELISA showed that both GFAP and S-100b decreased in the olfactory cortex, the hypothalamus, the thalamus, and the hippocampus of the 3-w group. The only effect observed in the 1 w group was a decrease of S-100b in the olfactory cortex at the highest dose. The immunohistochemistry showed no noticeable reduction in the number of astrocytes. We assume that the decreased levels of GFAP and S-100b are due to an adverse effect of Mn on the astrocytes, although this effect does not result in astrocytic demise. In the 3-w group, exposed to the highest dose of Mn, increased levels of GFAP and S-100b were observed in the olfactory bulbs, but these effects are probably secondary to a Mn-induced damage of the olfactory epithelium. Our results indicate that the astrocytes are the initial targets of Mn toxicity in the CNS. PMID- 10828273 TI - Pubertal development and reproductive functions of Crl:CD BR Sprague-Dawley rats exposed to bisphenol A during prenatal and postnatal development. AB - Bisphenol A (BPA) is used on a large scale in the manufacture of polycarbonate plastics. BPA has been shown to bind weakly to both estrogen receptor (ER)alpha and ERbeta, and to transactivate reporter genes in vitro. The purpose of the present study was to determine whether exposure of rats to BPA during pre- and postnatal development affects estrogen-mediated end points related to pubertal development and reproductive functions. BPA was administered to pregnant Crl:CD BR Sprague-Dawley rats by gavage at 0, 3.2, 32, or 320 mg/kg/day from gestation day (GD) 11 through postnatal day (PND) 20. Diethylstilbestrol (DES) at 15 microg/kg/day was used as a reference chemical with known estrogenic effects. Female pubertal development was not affected by indirect BPA exposure of the offspring at any of the dose levels. Treatment with this chemical also did not produce detectable effects on the volume of the sexually dimorphic nucleus of the preoptic area (SDN-POA), estrous cyclicity, sexual behavior, or male reproductive organ weights of F(1) offspring. However, DES at 15 microg/kg/day increased the volume of the SDN-POA of female offspring and affected their normal estrous cyclicity following puberty. In this study, pre- and postnatal exposure of rats to BPA at 3.2, 32, or 320 mg/kg/day from GD 11 through PND 20 did not have any apparent adverse effects on female rat pubertal development and reproductive functions. PMID- 10828274 TI - Prenatal toxicity and lack of carcinogenicity of 2-amino-3-methylimidazo[4,5 f]quinoline (IQ) following transplacental exposure. AB - Accumulating evidence from human and experimental animal studies indicates that consumption of heterocyclic amines (HA), derived from cooked meat and fish, may be associated with an increased incidence of cancer. Experiments were initiated to assess the role of one of these compounds, 2-amino-3-methylimidazo[4,5 f]quinoline (IQ), as a potential transplacental carcinogen, as well as to evaluate whether in utero exposure to IQ results in the induction of fetal cytochrome P4501A1 (Cyp1a1), P4501B1 (Cyp1b1), and/or glutathione S-transferase (GST). Inducible, or responsive, backcrossed fetuses resulting from a cross between congenic C57BL/6 (Ah(d)Ah(d)) nonresponsive female mice and C57BL/6 (Ah(b)Ah(b)) responsive male mice were transplacentally exposed to olive oil or 6.25, 12.5, or 25 mg/kg of IQ on day 17 of gestation. No macroscopically or microscopically visible liver, lung, or colon tumors were found in the transplacentally treated offspring by one year after birth. Ethoxyresorufin O deethylase (EROD) and 1-chloro-2,4-dinitrobenzene assays were performed to evaluate whether transplacental exposure to IQ results in the induction of fetal Cyp1a1 and GST, respectively, in lung and liver tissues. Results showed levels of EROD and GST activity in tissues of IQ-treated mice to be very close, if not identical, to those of mice treated with olive oil. Similarly, ribonuclease protection assay data showed that the levels of Cyp1a1 and Cyp1b1 RNA in tissues of IQ-treated mice were not significantly different from those of oil-treated controls. Previous studies have shown that the developing organism expresses very low levels of Cyp1a2. Thus, in utero exposure to IQ does not lead to induction of Cyp1a1, Cyp1a2, or Cyp1b1 in the fetal compartment, thereby maintaining the low levels of these activating enzymes in the developing organism. Taken together, these data imply that, at least under the conditions employed for these experiments, IQ may not play an important role in transplacentally induced tumorigenesis. PMID- 10828275 TI - Bioassays of shortened duration for drugs: statistical implications. AB - Declining survival rates in rodent carcinogenesis bioassays have raised a concern that continuing the practice of terminating such studies at 24 months could result in too few live animals at termination for adequate pathological evaluation. One option for ensuring sufficient numbers of animals at the terminal sacrifice is to shorten the duration of the bioassay, but this approach is accompanied by a reduction in statistical power for detecting carcinogenic potential. The present study was conducted to evaluate the loss of power associated with early termination. Data from drug studies in rats were used to formulate biologically based dose-response models of carcinogenesis using the 2 stage clonal expansion model as a context. These dose-response models, which were chosen to represent 6 variations of the initiation-promotion-completion cancer model, were employed to generate a large number of representative bioassay data sets using Monte Carlo simulation techniques. For a variety of tumor dose response trends, tumor lethality, and competing risk-survival rates, the power of age-adjusted statistical tests to assess the significance of carcinogenic potential was evaluated at 18 and 21 months, and compared to the power at the normal 24-month stopping time. The results showed that stopping at 18 months would reduce power to an unacceptable level for all 6 submodels of the 2-stage clonal expansion model, with the pure-promoter and pure-completer models being most adversely affected. For the 21-month stopping time, the results showed that, unless pure promotion can be ruled out a priori as a potential carcinogenic mode of action, the loss of power is too great to warrant early stopping. PMID- 10828276 TI - Chronic toxicity of di(2-ethylhexyl)phthalate in rats. AB - Fischer 344 rats were treated with 0, 100, 500, 2500, or 12,500 ppm di(2 ethylhexyl)phthalate (DEHP) in the diet for up to 104 weeks. Blood and urine were analyzed at weeks 26, 52, 78, and 104 from 10 animals per sex per group. Survival was slightly but not statistically reduced for rats receiving 12,500 ppm DEHP. Body weights and food consumption were significantly reduced for rats receiving the highest dose level of DEHP and occasionally for the male 2500-ppm group. BUN and albumin were significantly higher and globulin lower at nearly every sampling interval for the 12,500-ppm group compared with the controls. There was an increase in the mean activities of AST and ALT at 104 weeks, but no statistically significant differences were seen. Erythrocyte count, hemoglobin, and hematocrit values for the 12,500-ppm group were significantly lower than controls at nearly every sampling interval. No other differences in hematology were seen. No toxicologically significant changes were observed in urinalysis. At termination, relative lung weights for the 2500- and 12,500-ppm male groups of rats were significantly higher than for the controls. Absolute and relative liver and kidney weights for the 2500- and 12,500-ppm male rats, and liver weights for 12,500-ppm female rats were higher compared with the controls. Absolute and relative testes weights for the 12, 500-ppm male rats were lower compared with the controls. All organs were examined for histopathology. The incidence of hepatocellular lesions has been reported separately and correlated with the induction of peroxisomal enzyme activity (David et al., 1999). A dose level of 500 ppm was the NOEL for peroxisome proliferation. Bilateral aspermatogenesis in the testes, castration cells in the pituitary gland, spongiosis hepatis, and pancreatic acinar cell adenoma were observed for 12,500-ppm male rats. Aspermatogenesis and spongiosis hepatis were observed for 2500-ppm male rats, and aspermatogenesis was seen at 500 ppm. DEHP exposure exacerbated age-, species- or strain-related lesions such as mineralization of the renal papilla and chronic progressive nephropathy in male rats. Kupffer cell pigmentation and renal tubule pigmentation were seen in male and female 12,500-ppm rats. The increased incidence of spongiosis hepatis correlated with increased palmitoyl CoA oxidase activity, but the incidence of pancreatic acinar cell adenoma was increased only at the highest dose level of 12,500 ppm. These lesions, although typical of those seen with other peroxisome proliferators, may respond differently depending on the potency of the peroxisome proliferator. A dose level of 500 ppm (28.9-36.1 mg/kg/day) was considered to be the NOAEL. PMID- 10828277 TI - Bacterial lipopolysaccharide exposure augments aflatoxin B(1)-induced liver injury. AB - Bacterial endotoxin (lipopolysaccharide; LPS) given to animals in large doses results in pronounced, midzonal liver injury. Exposure to smaller, non-injurious doses of LPS augments the toxicity of certain hepatotoxicants. This study was conducted to delineate the development of injury in a rat model of augmentation of aflatoxin B(1) (AFB(1)) hepatotoxicity by LPS. At large doses (i.e., > 1 mg/kg, ip), AFB(1) administration resulted in pronounced injury to the periportal regions of the liver. Male, Sprague-Dawley rats (250-350 g) were treated with 1 mg AFB(1)/kg, ip or its vehicle (0.5% DMSO/saline) and 4 h later with either E. coli LPS (7.4 x 106 EU/kg, iv) or its saline vehicle. Liver injury was assessed 6, 12, 24, 48, 72, or 96 h after AFB(1) administration. Hepatic parenchymal cell injury was evaluated as increased alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities in serum and from histologic examination of liver sections. Biliary tract alterations were evaluated as increased concentration of serum bile acids and activities of gamma-glutamyltransferase (GGT), alkaline phosphatase (ALP), and 5'-nucleotidase (5'-ND) in serum. At all times and for all markers, injury in rats treated with either AFB(1) or LPS alone was absent or modest. In the AFB(1)/LPS cotreated group, hepatic parenchymal cell injury was pronounced by 24 h and had returned to control values by 72 h. The injury began in the periportal region and spread midzonally with time. Furthermore, changes in serum markers indicative of biliary tract alterations were evident by 12 h and had returned to control values by 72 h. Thus, the nature of the hepatic lesions suggested that LPS potentiated the effects of AFB(1) on both parenchymal and bile duct epithelial cells. PMID- 10828278 TI - Effects of cartap on isolated mouse phrenic nerve diaphragm and its related mechanism. AB - Cartap, a nereistoxin analogue pesticide, is reported to have no irritation to eyes in rabbits. However, we have demonstrated recently that cartap could actually cause acute death in rabbits via ocular exposure. Our preliminary study with isolated mouse phrenic nerve diaphragms has shown that instead of neuromuscular blockade, cartap caused muscular contracture. The objective of the study was to examine the effect of cartap on the neuromuscular junction in more detail and to investigate its possible underlying mechanism with isolated mouse phrenic nerve diaphragms and sarcoplasmic reticulum (SR) vesicles. Cartap or nereistoxin at various concentrations was added in the organ bath with isolated mouse phrenic nerve diaphragm and both nerve- and muscle-evoked twitches were recorded. Instead of blocking the neuromuscular transmission as nereistoxin did, cartap caused contracture in stimulated or quiescent isolated mouse phrenic nerve diaphragm. Both the cartap-induced muscular contracture force and the time interval to initiate the contracture were dose-dependent. The contracture induced by cartap was not affected by the pretreatment of the diaphragm with the acetylcholine receptor blocker alpha-bungarotoxin; the Na(+) channel blocker tetrodotoxin; or various Ca(2+) channel blockers, NiCl(2), verapamil, and nifedipine. On the contrary, the contracture was significantly inhibited when the diaphragm was pretreated with ryanodine or EGTA containing Ca(2+)-free Krebs solution or in combination. This suggested that both internal and extracellular Ca(2+) might participate in cartap-induced skeletal muscle contracture. Moreover, cartap inhibited the [(3)H]-ryanodine binding to the Ca(2+) release channel of SR in a dose-dependent manner. Additionally, cartap could induce a significant reduction in Ca(2+)-ATPase activity of SR vesicles at a relatively high dose. The results suggested that cartap might cause the influx of extracellular Ca(2+) and the release of internal Ca(2+), with subsequent induction of muscular contracture in the isolated mouse phrenic nerve diaphragm. Based on these findings, we propose that the acute death of rabbits following ocular exposure to cartap might have resulted from respiratory failure secondary to diaphragm contracture. PMID- 10828279 TI - Metallothionein-I/II null mice are more sensitive than wild-type mice to the hepatotoxic and nephrotoxic effects of chronic oral or injected inorganic arsenicals. AB - Metallothionein (MT) is a low-molecular-weight, sulfhydryl-rich, metal-binding protein that can protect against the toxicity of cadmium, mercury, and copper. However, the role of MT in arsenic (As)-induced toxicity is less certain. To better define the ability of MT to modify As toxicity, MT-I/II knockout (MT-null) mice and the corresponding wild-type mice (WT) were exposed to arsenite [As(III)] or arsenate [As(V)] either through the drinking water for 48 weeks, or through repeated sc injections (5 days/week) for 15 weeks. Chronic As exposure increased tissue MT concentrations (2-5-fold) in the WT but not in MT-null mice. Arsenic by both routes produced damage to the liver (fatty infiltration, inflammation, and focal necrosis) and kidney (tubular cell vacuolization, inflammatory cell infiltration, and interstitial fibrosis) in both MT-null and WT mice. However, in MT-null mice, the pathological lesions were more frequent and severe when compared to WT mice. This was confirmed biochemically, in that, at the higher oral doses of As, blood urea nitrogen (BUN) levels were increased more in MT-null mice (60%) than in WT mice (30%). Chronic As exposures produced 2-10 fold elevation of serum interleukin-1beta, interleukin-6, and tumor necrosis factor alpha levels, with greater increases seen by repeated injections than by oral exposure, and again, MT-null mice had higher serum cytokines than WT mice after As exposure. Repeated As injections also decreased hepatic glutathione (GSH) by 35%, but GSH-peroxidase and GSH-reductase were minimally affected. MT-null mice were more sensitive than WT mice to the effect of GSH depletion by As(V). Hepatic caspase-3 activity was increased (2-3-fold) in both WT and MT-null mice, indicative of apoptotic cell death. In summary, chronic inorganic As exposure produced injuries to multiple organs, and MT-null mice are generally more susceptible than WT mice to As-induced toxicity regardless of route of exposure, suggesting that MT could be a cellular factor in protecting against chronic As toxicity. PMID- 10828280 TI - The effects of sulfur, thiol, and thiol inhibitor compounds on arsine-induced toxicity in the human erythrocyte membrane. AB - The mechanism of arsine (AsH(3)) toxicity is not completely understood. The first cytotoxic effect of AsH(3) is disruption of ion homeostasis, with a subsequent hemolytic action. The only accepted treatment for AsH(3) toxicity is exchange transfusion of the blood. In this study the effect of sulfur, sulfur compounds, thiol-containing compounds, and thiol inhibitors on AsH(3)-induced disruption of membrane transport and hemolysis in human erythrocytes was investigated in vitro. Elemental sulfur, sodium thiosulfate, 5, 5'-dithio-bis(2-nitrobenzoic acid), and meso-2,3-dimercaptosuccinic acid were successful in delaying hemolysis, but the most successful agent was the sulfhydryl inhibitor, N-ethylmaleimide (NEM). This indicated that sulfhydryl groups, possibly membrane sulfhydryls, are major factors in the hemolytic mechanism of AsH(3). Measuring intracellular ion concentrations tested the effect of NEM on AsH(3)-induced disruption of membrane transport. AsH(3) alone caused all ions tested to flow with their concentration gradients: Intracellular K+ and Mg++ decreased, whereas Na+, Cl-, and Ca++ increased. NEM was unable to prevent ion loss except for Ca++, whose increase was prevented for 1 h after AsH(3) treatment. The influx of Ca++ in AsH(3)-treated erythrocytes is an irreversible event leading to hemolysis. Reduction of oxygenated hemoglobin to carboxyhemoglobin completely inhibited AsH(3)-induced hemolysis. In addition, AsH(3) and NEM had no direct chemical interactions. We concluded that membrane sulfhydryl groups are likely targets of AsH(3) toxicity, with NEM being able to prevent AsH(3)-induced hemolysis. PMID- 10828281 TI - Mobilization of vitamin A stores in rats after administration of 2,3, 7,8 tetrachlorodibenzo-p-dioxin: a kinetic analysis. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a highly toxic environmental contaminant that prevents the normal accumulation of vitamin A in liver and causes increased excretion of vitamin A. To determine what alterations in vitamin A metabolism occur first in response to TCDD treatment, we administered TCDD (7.0 microg/kg b.w. ) orally to rats that had received a nonperturbing (tracer) iv dose of [(3)H]vitamin A-labeled plasma (n = 3) or lymph (n = 3) 21 days earlier. Within a few days after TCDD administration, fraction of the injected radiolabel in plasma, which had been in a terminal slope when plotted on a semilog scale, increased and remained elevated until the experiment was terminated (day 42). At that time, liver vitamin A levels were 65% lower in TCDD-perturbed rats than in controls. Using model-based compartmental analysis and compartmental models developed previously for control rats (S. K. Kelley et al., 1998, Toxicol. Sci, 44:1-13), we determined the minimal changes needed to account for the perturbation in plasma [(3)H] tracer responses after TCDD administration. We determined that the effects of TCDD could be explained by adjusting the value of one fractional transfer coefficient corresponding to the mobilization of vitamin A from large, slowly turning-over pools. We speculate that this change corresponds to an increased fractional rate of retinyl ester hydrolysis, and that it precedes the TCDD-associated increased irreversible utilization and excretion of vitamin A. PMID- 10828287 TI - Exploring the nature of molecular recognition in nicotinic acetylcholine receptors. AB - Nicotinic acetylcholine receptors (nAChRs) are the subject of ever increasing interest because of their presumed involvement in the etiology of numerous clinical disorders. Unfortunately, the absence of atomic-level structural data, as well as the pharmacological complexity of these receptors leaves many fundamental questions unanswered. An understanding of how ligands interact with the receptor and, in-turn, how these interactions lead to pharmacological effect is vital in the advancement of nAChR-based therapeutics. We will first explore physico-chemical themes that are evidenced to be of particular importance in nAChR molecular recognition; these are- pi-cation interaction, conformational entropy and stereochemistry. The second objective of this review is an interpretive encapsulation of the extensive and disparate body of structure activity data that now exists for nAChRs. Finally, this review will advocate a re investigation of distance geometry based methods as well as the need for additional approaches in nicotinic receptor pharmacophore generation. PMID- 10828288 TI - Inhibition of transglycosylation involved in bacterial peptidoglycan synthesis. AB - The continuing spectre of resistance to antimicrobial agents has driven a sustained search for new agents that possess activity on drug resistant bacteria. Although several paths are available to reach this goal, the most generalized would be the discovery and clinical development of an agent that acts on a new target which has not yet experienced selective pressure in the clinical setting. Such a target should be essential to the growth and survival of bacteria, and sufficiently different from, or better still non-existent in, the human host. The transglycosylation reaction that polymerizes biochemical intermediates into peptidoglycan qualifies as such a target. This biochemical system accepts the basic unit N-acetylglucosamine-beta-1, 4-N-acetyl-muramyl-pentapeptide pyrophosphoryl-undecaprenol (lipid II), and leads to polymerization of the N acetylglucosamine -beta-1, 4-N-acetyl-muramyl-pentapeptide segment into peptidoglycan. Approaches to targeting this reaction include modification of known glycolipid and glycopeptide natural product antibiotics. The synthesis and antibacterial activity of synthetic analogs of moenomycin having novel antibacterial activities not present in the parent structure will be presented, together with the combinatorial chemistry and assay systems leading to their discovery. Likewise, we will discuss chemical modifications to specific glycopeptide antibiotics that have extended their spectrum to include vancomycin resistant enterococci that substitute D-alanyl-D-lactate for D-alanyl-D-alanine in their peptidoglycan. Two differing theories, one positing the generation of high affinity, specific binding to D-alanyl-D-lactate via glycopeptide dimerization and/or membrane anchoring, and the other supporting direct targeting of the modified glycopeptide to the transglycosylation complex, seek to explain the mechanism of action on vancomycin resistant enterococci. Biochemical evidence in support of these two theories will be discussed. PMID- 10828289 TI - New chemical and biological aspects of S-nitrosothiols. AB - Nitric oxide (NO) possesses many physiological effects and S-nitrosothiols have been identified in a variety of tissues exhibiting many NO-like activities. This review focuses on the latest discoveries pertaining to the biological functions of S-nitrosothiols and the recent research progress in the chemical properties and biomedical applications of RSNOs. PMID- 10828290 TI - Target-based drug discovery for malaria, leishmaniasis, and trypanosomiasis. AB - Advances in combinatorial chemistry, high-throughput screening, and molecular modeling have revolutionized the process of drug discovery in the pharmaceutical industry. Drug discovery efforts for the primary protozoal parasitic diseases of the developing world, malaria, leishmaniasis, and trypanosomiasis, have also begun to employ these techniques. Drug targets in these parasites, exemplified by cysteine proteases and trypanothione reductase, have been purified and used for inhibitor screening. Through this work, small molecules have been identified that inhibit both parasite proteins and the growth of the organisms. This review describes advances that have been made in examining the effects of small molecules on potential parasitic drug targets determined by biochemical and computer-based screening, and also details the activity of such compounds on parasites in vitro and in vivo. Based on these results, it is apparent that modern drug discovery techniques hold promise for the identification of antiparasitic drug candidates. PMID- 10828297 TI - Non-invasive radiotracer imaging as a tool for drug development. AB - Non-Invasive Radiotracer Imaging (NIRI) uses either short-lived positron-emitting isotopes, such as 11C and 18F, for Positron Emis ion Tomography (PET) or single photon emitting nuclides, e.g., 123I, which provide images using planar imaging or Single-Photon Emission Computed Tomography (SPECT). These high-resolution imaging modalities provide anatomical distribution and localization of radiolabeled drugs, which can be used to generate real time receptor occupancy and off-rate studies in humans. This can be accomplished by either isotopically labeling a potential new drug (usually with 11C), or indirectly by studying how the unlabelled drug inhibits specific radioligand binding in vivo. Competitive blockade studies can be accomplished using a radiolabeled analogue which binds to the site of interest, rather than a radiolabeled version of the potential drug. Imaging, particularly PET imaging, can be used to demonstrate the effect of a drug through a biochemical marker of processes such as glucose metabolism or blood flow. NIRI as a development tool in the pharmaceutical industry is gaining increased acceptance as its unique ability to provide such critical information in human subjects is recognized. This section will review recent examples that illustrate the utility of NIRI, principally PET, in drug development, and the potential of imaging advances in the development of cancer drugs and gene therapy. Finally, we provide a brief overview of the design of new radiotracers for novel targets. PMID- 10828298 TI - Bioanalytical applications of accelerator mass spectrometry for pharmaceutical research. AB - Accelerator mass spectrometry (AMS) is a mass spectrometric method for quantifying isotopes. It has had great impact in the geosciences and is now being applied in the biomedical fields. AMS measures radioisotopes such as 14C, 3H, 41Ca, and 36Cl, and others, with attomole sensitivity and high precision. Its use is allowing absorption, distribution, metabolism and elimination studies, as well as detailed pharmacokinetics, to be carried out directly in humans with very low chemical or radiological hazard. It is used in combination with standard separation methodologies, such as chromatography, in identification of metabolites and molecular targets for both toxicants and pharmacologic agents. AMS allows the use of very low specific activity chemicals (< 1 mCi/mmol), creating opportunities to use compounds not available in a high specific activity form, such as those that must be biosynthesized, produced in combinatorial libraries, or made through inefficient synthesis. AMS is allowing studies to be carried out with agents having low bioavailability, low systemic distributions, or high toxicity where administered doses must be kept low (<1 microg/kg). It may have uses in tests for idiosyncratic metabolism, drug interaction, or individual susceptibility, among others. The ability to use very low chemical doses, low radiological doses, small samples and conduct multiple dose studies may help move drug candidates into humans faster and safer than before. The uses of AMS are growing and its potential for drug development is only now beginning to be realized. PMID- 10828299 TI - Recent advances in the applications of radioisotopes in drug metabolism, toxicology and pharmacokinetics. AB - Radioisotopes have proven to be an indispensable tool in biomedical research and have played a pivotal role in the investigation of absorption, distribution, metabolism and excretion (ADME) properties of new chemical entities over the past several decades. The main advantage of using radioisotopes in studying the disposition of new drug candidates is the ease of detection and the achievement of high sensitivity, especially when compounds with high specific activity are used. The recent advances and applications of radioisotopes in designing and conducting ADME studies and its impact in the field of drug metabolism and pharmacokinetics are discussed in this review. PMID- 10828300 TI - Recent developments in tritium incorporation for radiotracer studies. AB - The essential nature of rapid radiotracer synthesis very early in drug discovery programs has driven the need for better and more varied tritium incorporation methods. This review presents a summary of recent advances for tritium introduction via tritiated water, tritium gas, complex tritides, and a range of recently improved tritiation reagents. Access to a wider range of tritiated reagents (for tritioacetylation, tritioformylation, methylation, etc.) and commercial manifolds for the transfer and use of tritium gas is also discussed. PMID- 10828301 TI - Recent advances in the design and synthesis of carbon-14 labelled pharmaceuticals from small molecule precursors. AB - Over the past decade, the increased chemical complexity of new drug candidates has resulted in a parallel need to develop innovative syntheses of carbon-14 labelled pharmaceuticals. Faced with short time-lines and a limited number of labelled precursors, radiochemists have addressed this challenge by developing new reagents and adapting existing technology to labelled syntheses. Selected examples from the recent radiochemical literature illustrate some of the creative strategies used to rapidly solve these synthetic challenges. Examples describing the handling and use of common small molecule reagents, such as carbon-14 labelled carbon dioxide, methyl iodide, cyanide, acetic acids, sulfur and phosphorous stabilized ylides for the synthesis of labelled steroids, prostanoids, nucleosides, pyridines, quinolines, benzazepines and other heterocycles are presented. Several general strategies for radiolabelling are also discussed including the degradation strategy for accessing necessary intermediates and precursors, the radiolabelling of aromatic substrates, transition metal mediated cross-couplings, and the use of chiral auxiliaries for the enantioselective syntheses of radiolabelled pharmaceuticals. PMID- 10828302 TI - Mechanisms for the selective actions of vitamin D analogues. AB - The wide range of activities now attributed to 1,25(OH)2D3 has suggested numerous potential therapeutic applications for this vitamin D hormone, including inhibiting growth of various type of cancer. Unfortunately, the potent calcemic activity of the natural hormone has precluded its use in most cases. Vitamin D analogs with higher therapeutic indices offer renewed hope for treatment of malignancies. The promising analogs currently under study were selected from hundreds of candidates by in vitro screening followed by in vivo testing. The mechanism(s) responsible for the greater effectiveness of most of these compounds is not known. Our current understanding of vitamin D physiology and biochemistry suggests that the biological profile of an analog would be determined primarily by its interaction with four classes of proteins: 1) the nuclear vitamin D receptor (VDR) that mediates transcriptional regulation; 2) the metabolic enzymes, primarily the vitamin D-24-hydroxylase but possibly others; 3) serum transporters, mainly vitamin D binding protein (DBP), and perhaps lipoproteins; and 4) a new class of receptors that reside in the plasma membrane and mediate rapid, nongenomic responses. This article discusses how the manner in which analogs associate with these proteins can potentially produce selective actions at the tissue, cell and gene level. A thorough understanding of the influence of these analog/protein interactions on the biological profile of vitamin D analogs will be invaluable for the design of future analogs with enhanced target specificity. PMID- 10828303 TI - Anti-tumor effects of 1,25-dihydroxyvitamin D3 and vitamin D analogs. AB - The role of 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) as a regulator of cell growth and differentiation is well recognized. Currently, 1, 25-(OH)2D3 and vitamin D analogs are being evaluated for their therapeutic potential in the treatment of hyperproliferative disorders like cancer. In the present review, we will discuss several processes that might be involved in 1,25-(OH)2D3- and vitamin D analog mediated suppression of cancer cell growth. The effects on tumor cell proliferation, differentiation, apoptosis, angiogenesis, metastases, and parathyroid hormone-related peptide secretion will be highlighted. In addition, combination therapy with other tumor effec tive drugs will be addressed. Furtermore, we will focus on the potential drawbacks and the possible side effects of vitamin D compounds in the treatment of cancer. PMID- 10828304 TI - Structure-function analysis of vitamin D and VDR model. AB - In the first section, the general three-dimensional structure of the ligand binding domain (LBD) of nuclear receptors (NR) was briefly described on the basis of their x-ray crystal structures. Emphasis was placed on the three major conformations of NR-LBD and their role in the transactivation function. In the second part, the structure-function relationship of vitamin D was analyzed based on the ligand structure, in particular by using systematic conformational analysis as a tool. On the basis of the conformational analysis of the vitamin D side chain and studies using conformationally restricted synthetic vitamin D analogs, we suggested the active space region concept of vitamin D: The vitamin D side-chain region was grouped into five regions (A, G, EA, EG and F). Activity orders, in terms of the spatial region, found by these studies are as follows: Affinity for vitamin D receptor (VDR), EA>A>F>G>EG; Affinity for vitamin D binding protein (DBP), A>>G,EA, EG; Target gene transactivation, EA>F>A>EG G; Cell differentiation, EA>F>A>EG G; Bone calcium mobilization, EA>G A>F EG; Intestinal calcium absorption, EA=A G>>EG. In the third section, homology modeling of VDR-LBD and docking of the natural ligand, 1,25-(OH)2D3, into the ligand binding cavity of the model are described. Amino acid residues forming hydrogen bonds with the biologically important 1alpha- and 25-OH groups were identified: 1alpha-OH forms a pincer-type hydrogen bond with R274 and S237 and 25 OH with H397. This VDR-LBD/1,25-(OH)2D3 docking model was firmly substantiated by mutation analysis. Using this VDR model, the structure-function relationship of highly potent vitamin D analogs was discussed. PMID- 10828305 TI - A novel class of aromatic vitamin D analogs. AB - Vitamin D analogs in which the triene moiety is replaced by an aromatic ring have been synthesized and their ability to bind to the vitamin D receptor investigated. PMID- 10828306 TI - Biological activity in vitro of side-chain modified analogues of calcitriol. AB - The results of our studies on the biological activity of side-chain modified analogues of vitamin D are reviewed. These analogues appeared to be effective in induction of cell differentiation, inhibition of tumour cell proliferation in vitro and in increasing of antitumour effect of cytostatics. On the other hand, inhibition of cytostatic-induced apoptosis by these compounds was observed. The mechanism of the antiproliferative effect of calcitriol analogues in vitro is discussed. The induction of antigens CD14 and CD11b expression and phagocytic activity of HL-60 cells after exposure to these compounds is related to their effect on cell differentiation. The differentiation of the HL-60 leukaemia cells induced by side-chain modified analogues of calcitriol increases their sensitivity to the antiproliferative effect of cisplatin, doxorubicin and genistein, despite of that this pretreatment causes resistance of these cells to cytostatics-induced apoptosis. We observed a synergistic antiproliferative effect of the combined therapy using analogues of calcitriol with subsequent treatment with the above-mentioned cytostatics. This effect was measured as a significant decrease of the ID50 values for each cytostatic applied after pretreatment of the tumour cells with the calcitriol analogues. The results presented suggest that the improved therapeutic effect may be achieved also in vivo by the combined application of the analogues (without calcemic activity) of calcitriol with antitumour agents. PMID- 10828307 TI - New synthetic vitamin D analogs with antiproliferative activities. AB - The introduction of oxygen atoms into different positions of the vitamin D side chain is described. By combining the arising 23-oxa and 25-oxa elements with other structural modifications (19-nor, iso-19-nor, 20-methyl, 20-ene, 20,21 cyclo) calcitriol analogs with remarkable levels of dissociation between beneficial acitivities on cell growth regulation and undesired hypercalcemia were identified. Structure-activity relations are elaborated in a very systematic outline of the Schering drug finding program in this particular class of vitamin D compounds. PMID- 10828308 TI - Biological activities of 19-nor-1alpha,25-dihydroxyvitamin D3 analogs singly dehydroxylated at the C-1 or C-3 position of the A-ring. AB - Growing interests have been focused on the development of hybrid-analogs with modifications of the A-ring and the side chain of 1alpha,25-dihydroxyvitamin D3 [1alpha,25(OH)2D3]. An exocyclic methylene group at C-10, a hydroxy group at C-1 and a hydroxy group at C-3 play a crucial role in the expression of biological activities of 1alpha,25(OH)2D3. However, relationship between the functional groups and activities has not been fully understood. We have synthesized and evaluated biological activities of several singly dehydroxylated A-ring analogs of 19-nor-1alpha,25(OH)2D3 and 19-nor-22-oxa-1alpha,25(OH)2D3. All of them have an extremely low binding affinity for vitamin D receptor (VDR). Some of them lack the 1alpha-hydroxy group that is considered to be essential for VDR-mediated gene expression, have greater or equivalent potencies to 1alpha,25(OH)2D3 for inducing differentiation and cell cycle G0-G1 arrest of human promyelocytic leukemia cells as well as for the transactivation of target genes including a rat 25 hydroxyvitamin D3-24-hydroxylase gene promoter and a human osteocalcin gene promoter in transfected mammalian cells. The assessment of a ligand/VDR/Retinoid X receptor complex formation using a two-hybrid luciferase assay revealed that the liganded VDR has high potency to form a heterodimer, but this could not explain the high biological potency of the 19-nor analogs. Other reason(s) including an interaction with transcriptional cofactors should be considered to explain the mechanism of action of 19-nor analogs. PMID- 10828309 TI - Seocalcitol (EB 1089): a vitamin D analogue of anti-cancer potential. Background, design, synthesis, pre-clinical and clinical evaluation. AB - It is well established that the metabolically active form of vitamin D, 1alpha,25 dihydroxyvitamin D3 (1alpha,25(OH)2D3) plays a key role in the establishment and maintenance of the calcium metabolism in the body. In addition to this classic effect of 1alpha,25(OH)2D3, substantial evidence has emerged demonstrating that 1alpha,25(OH)2D3 is able to regulate cell growth and differentiation in a number of different cell types, including cancer cells. However, the clinical usefulness of 1alpha,25(OH)2D3 is limited by its tendency to cause hypercalcaemia. Much effort has therefore been directed to identifying new vitamin D analogues with potent cell regulatory effects, but with weaker effects on the calcium metabolism than those of 1alpha,25(OH)2D3. One of these new synthetic analogues is Seocalcitol (EB 1089). Despite being 50-200 times more potent than 1alpha,25(OH)2D3 with respect to regulation of cell growth and differentiation in vitro as well as in vivo, EB 1089 displays a reduced calcaemic activity in vivo compared to that of 1alpha, 25(OH)2D3. These characteristics make EB 1089 a potentially useful compound for the treatment of cancer. Recent clinical evaluation of EB 1089 has focused mainly on establishing a maximum tolerated dose in cancer patients. Early results confirm that the low calcaemic activity observed in animals can be reproduced in the clinic. Furthermore, EB 1089 has been shown to induce regression of tumours, especially in hepatocellular carcinoma where complete remission has been obtained. In conclusion, the development of EB 1089 as an anti-cancer drug holds promise. However, its final evaluation must await the completion of ongoing controlled clinical trials. PMID- 10828310 TI - The new antiepileptic drugs: pharmacological and clinical aspects. AB - In recent years several new drugs (oxcarbazepine, lamotrigine, topiramate, gabapentin, zonisamide, tiagabine, fosphenytoin, vigabatrin and felbamate) have been added to the therapeutic armamentarium against epilepsy. Some of these represent structural modifications of pre-existing compounds, others were developed with the specific objective of modifying neurotransmitter function, and many more were found to be clinically useful even though their mode of action is unclear or differs from that originally planned. The pharmacokinetics of these drugs differ widely from one agent to another. Some (gabapentin and vigabatrin) are eliminated unchanged in urine and have little or no interaction potential; others (tiagabine, lamotrigine, topiramate, oxcarbazepine, zonisamide, felbamate) are subject to induction of metabolism by concomitant anticonvulsants; lamotrigine is vulnerable to metabolic inhibition by valproate, and felbamate is a powerful enzyme inhibitor in addition to being an inducer of the metabolism of carbamazepine and steroid oral contraceptives. All new antiepileptic drugs have been found to be effective in improving seizure control in patients with partial and secondarily generalized seizures. However, lamotrigine, topiramate, zonisamide and felbamate appear to have broader efficacy against both partial and many generalized seizure types, while vigabatrin is also valuable in the management of infantile spasms. In monotherapy studies, new drugs have not been found to be more efficacious than older agents, but some may offer limited advantages in terms of improved tolerability. On the other hand, serious toxicity restricts considerably the use of vigabatrin and felbamate. Overall, new drugs represent valuable tools in the fight against epilepsy, but because of limited experience and cost considerations their first-line use cannot be recommended in most situations. PMID- 10828311 TI - Stopping drug therapy in epilepsy. AB - In most of the epilepsies and epileptic syndromes, the decision to initiate antiepileptic drug (AED) therapy is often a far simpler one than the decision to stop it. The primary factor that drives a patient to want to discontinue therapy, and a doctor to endorse or recommend this, is a fear of long-term adverse events, a consideration that may be entirely justified as the side effects of vigabatrin and felbamate have proved. On the other hand, seizure recurrence with the attendant implications in employment, driving regulations and social stigmatization is a strong deterrent that discourages withdrawal of therapy. The absence of a clearer understanding of the natural history and prognosis of many individual epilepsy syndromes somewhat hampers the resolution of this dilemma. Hopefully, as our understanding of the epilepsies grows and the prediction of the chances of seizure relapse becomes a more precise science, stopping medication will become a less fraught exercise for both the patient and the doctor. PMID- 10828312 TI - Current issues on epileptic women. AB - Issues linked to epileptic women are being reviewed. Ovarian steroid hormones have a number of effects on the brain that predispose to epileptic activity. In particular, estradiol produces changes in the hippocampus synapses predisposing hyperexcitability associated with seizures. Also, menses and menopause periods, in which there are changing levels of steroid ovarian hormones, are associated with a particular appearing of seizures (catamenial epilepsy) and with phenotypic changes of previous ones. Epilepsy can affect the reproductive system, inducing endocrinal abnormalities (through disruption of cortical regulation of hypothalamus hormone release, and changes in the central nervous system concentration of steroid hormones induced by antiepileptics), infertility (linked to abnormalities in menstrual cycle or to the occurrence of polycytic ovaries, particularly in association with valproate treatment), and sexual disfunction (namely related to physiologic defects). Oral hormonal contraceptives should be performed using a pill with > 50mg of estrogen in order to prevent its potential loss of efficacy induced by enzyme-inducing antiepileptics. Concerning pregnancy, some topics should be discussed with, and advised to epileptic women, including: the possibility of withdrawal antiepileptics and the need of folic acid supplementation when planning a pregnancy; the risk of increased seizure frequency during pregnancy, and of the occurrence of obstetric complications; the increased risk of teratogenesis associated with antiepileptic therapy (mainly if in polytherapy); the need of vitamin K supplementation during the last month of pregnancy in order to avoid newborn haemorrhages; and the general absence of risk of breastfeeding even under sustained antiepileptic therapy. PMID- 10828313 TI - The new drugs and the strategies to manage epilepsy. AB - After a short historical review of the development of the pharmaceutical treatment of the epilepsies the author reviews some of the possible strategies to manage patients with the different types of epilepsies and epileptic syndromes using the classical drugs. A strategy used by most of the physicians uses Sodium Valproate as the first line drug for almost all patients. This may be replaced by other drugs according to their efficacy against the different types of seizures to be treated whenever VPA has not enough efficacy or isn t well tolerated. On the other hand epileptologists use the different drugs according to the different epilepsies and epileptic syndromes depending on the relative efficacy of each drug available and the possible side effects. He then describes succinctly the better-known new drugs and makes some comments on the coming drugs now in development. Finally he proceeds to include them in the strategies above described. Lamotrigine and possibly Topiramate are good candidates to replace VPA in the one drug strategy. Lamotrigine, Oxcarbamazepine and possibly Gabapentin may be used in the future as 1st line drugs in selected patients. Vigabatrin is already used as one of the better alternatives for West syndrome and Oxcarbamazepine has replaced Carbamazepine in countries where it s available to the public. Some drawbacks have been apparent with these drugs like the hepatic and haematological toxic effect of Felbamate or the apparently irreversible fields constriction provoked by Vigabatrin, which did limit their use. PMID- 10828314 TI - Overview of childhood epilepsy and epileptic syndromes and advances in therapy. AB - Seizures have a variety of etiologies and manifestations. Descriptions of various epiletic seizures as well as electroencephalographic findings have led to a unifying international classification of epileptic seizures and epilepsy syndromes. The development of this classification system and the emergence of several new antiepiletic drugs have led to progress in the refractory pediatric patient particularly disorders which are traditionally difficult to treat such as infantile spasms and the Lennox-Gastaut Syndrome. However, there is limited data regarding optimal use in children. The childhood epilepsy syndromes are reviewed as well as the newer antiepileptic drug treatments - felbamate, gabapentin, lamotrigine, levetiracetam, oxcarbazepine, tiagabine, topiramate, and zonisamide. Efficacy data and toxicity are discussed from both the adult, and when available, pediatric data. PMID- 10828315 TI - Current status and recent developments in anthracenone antipsoriatics. AB - Treatment of psoriasis is rapidly changing, and there have been dramatic additions such as topical vitamin D analogs and a topical retinoid to the therapeutic armamentarium in recent years. Even though drug therapy is often inadequate and serves only to abate the skin disorder, anthralin of the anthracenone antipsoriatics clears the rash totally. Of all antipsoriatic agents, anthralin has proven to be the most remarkably consistent and time-honored drug for treating the disease. However, anthralin therapy is accompanied by inflammation and staining of the nonaffected skin surrounding a psoriatic lesion. A large body of evidence has shown that the biochemical basis for its mechanism of action at the molecular level is related to its redox chemistry leading to the production of oxygen radicals. These can react with all classes of biological macromolecules and are involved in the principal cellular effects of the anthracenones. This article will focus on current strategies to minimize the potential of skin inflammation by anthralin and the underlying concepts of controlling and manipulating oxygen-radical generation which are essential for the rational design of novel anthracenones. Various studies have shown that by altering the structure of anthralin the in vitro profile can be markedly altered from an only moderate 5-lipoxygenase inhibitor to a more potent inhibitor of this enzyme, which may reflect improved activity against the inflammatory component of psoriasis while the antiproliferative activity is retained. The structural change is also accompanied by significantly diminished oxygen-radical generation and proinflammatory action on the skin. In particular, derivatives bearing a phenylacyl substituent in the critical 10-position of the anthracenone molecule have a favorable overall profile for achieving improved topical therapy of psoriasis. PMID- 10828316 TI - Recent developments in receptor-selective retinoids. AB - Natural (all trans-retinoic acid, RA) and synthetic retinoids exhibit potent anti proliferative, normalization of differentiation and anti-inflammatory activities which appear to account for their therapeutic effects in acne, psoriasis, photoaging, precancerous lesions and established cancers. Although RA has shown considerable promise in dermatologic indications, certain side effects have restricted its use as a choice of agent for chronic administration. Systematic synthesis of receptor-selective retinoids has resulted in two topical drugs, Tazorac/Zorac (tazarotene) and Differin (adapalene). Tazorac is indicated for psoriasis and acne and Differin gel for the treatment of acne. These drugs bind to the retinoic acid receptor (RAR) family members. Various RAR subtype-specific and function-selective retinoids have been synthesized. These retinoids, which are in various stages of pre-clinical development for the treatment of cancers, psoriasis and as an antidote to Accutane-mediated mucocutaneous toxicity, will also be discussed in this review. Discovery of another retinoid receptor, retinoid X receptor (RXR), revealed that RXR-specific retinoids already existed in retinoid chemical libraries. Structure activity relationship studies based upon binding and transactivation assays led to the synthesis of RXR-specific ligands with high affinities for RXR subtypes. These compounds were found to be effective in the treatment of hyperglycemia in animal models of type II diabetes. The discovery of novel retinoids along with an increased understanding of the biological functions and mechanisms of action of retinoid receptors are likely to result in improved treatments for existing responsive indications and identification of new retinoid therapeutic targets. PMID- 10828317 TI - Inhibitors of tyrosine kinases in the treatment of psoriasis. AB - Psoriasis is a heterogenous skin disease, characterized by epidermal hyperproliferation, abnormal keratinization and inflammation. The heterogeneity of the disease results probably from the interaction of multiple gene abnormalities with environmental factors. The new approaches to drug design have become refocused to the emerging understanding of the role of signaling pathways in health and disease. Protein tyrosine kinases (PTKs) regulate cell proliferation, differentiation and immune processes. Uncontrolled signaling from receptor and intracellular tyrosine kinases can lead to numerous proliferative diseases: cancer, leukemia, restenosis and psoriasis. Identification of PTKs that play a key role in a defined disease can lead to a selective drug. The balance of signals which regulate the homeostasis of normal epidermis is altered in psoriasis. Several lines of evidence suggest a role for the EGF receptor system in this process. Therefore, blockers of the EGFR kinase were suggested as potent antipsoriasis agents. PTK inhibitors from the tyrphostin family were found to block EGF - dependent cell proliferation. AG 1571 (SU 5271) potently inhibits ligand-induced autophosphorylation of EGF-R, downstream signal transduction events, DNA replication and cell cycle progression at micromolar concentrations, as well as proliferation of keratinocytes isolated from psoriatic lesions in excellent correlation with its EGFR kinase inhibitory activity in these cells. AG 1571 (SU 5271) has been in clinical trials by SUGEN Inc. since early 1997. Overexpression of the EGFR is the hallmark of most epithelial cancers. Therefore one can view blockers of the EGFR kinase as becoming universal inhibitors. Tyrphostins are the first signal transduction agents to be used in the clinic. This article summarizes recent progress in the development of PTK inhibitors in the treatment of psoriasis. PMID- 10828318 TI - Inhibitors of the enzyme purine nucleoside phosphorylase as potential therapy for psoriasis. AB - Purine nucleoside phosphorylase (PNP) is one of the enzymes comprising the purine salvage pathway , and is responsible for the catalysis of the reversible phosphorolytic cleavage of purine ribonucleosides and 2'-deoxyribonucleosides. The pivotal role of PNP in T-cell proliferation has been demonstrated in patients with inherited PNP deficiency, where T-cell levels may be 1-3% of normal. This observation helped establish the critical role of PNP in T-cells and provided a rationale for developing inhibitors of PNP. Inhibitors of PNP may be useful for treating a variety of T-cell related autoimmune diseases including psoriasis, rheumatoid arthritis and Crohn s disease and T-cell cancers. In this manuscript, the x-ray crystal structure of the PNP enzyme is described. Results of a structure-based drug design program aimed at designing small-molecule inhibitors of PNP are also described. Of the many classes of compounds synthesized, studied and reviewed, only one, the 3-pyridinylmethyl-9-deazaguanine (BCX-34, 39) analog has been used in clinical trials. Both topical and oral formulations of BCX-34 were studied in psoriatic patients and the results of these clinical trials are described. PMID- 10828319 TI - Recent developments in vitamin D analogs. AB - Within the past decade it has been shown that psoriasis can be treated topically with analogs of vitamin-D3. Impaired differentiation and increased proliferation of keratinocytes are key features in psoriatic lesions together with a local activation of T lymphocytes. Evidence has accumulated showing that analogs of vitamin D3 increase differentiation and inhibit proliferation of keratinocytes. Therefore, analogs of vitamin D3 have been investigated in a number of trials showing improvement of psoriasis. It has been shown that vitamin D analogs are better than their vehicle and show the same potency as potent topical steroids. However, vitamin D analogs have been proven efficacious and without side effects also when used on long term basis. Vitamin D analogs can be used both as monotherapy and in combination topical steroids, UVB, PUVA, retinoids and cysclosporine. The vitamin D3 analog calcipotriol has been investigated in most detail and is available as an ointment, a creme and as a scalp solutation. From clinical studies involving thousands of patients, it can be concluded that calcipotriol is efficacious, safe, well tolerated and can be used on a long term basis. Other analogs are available, however, these analogs have not been studied in greater details yet. PMID- 10828320 TI - Do quadrupeds require a change in trunk posture to walk backward? AB - Previous studies on cats walking backward have indicated that they adopt a presumably adaptive posture characterized by extreme dorsiflexion of the lumbar spine. Because humans do not show any marked postural changes during backward walking, we questioned whether the posture exhibited by cats during backward walking was in fact adaptive and whether it was typical of quadrupeds. We therefore compared forward and backward walking in three treadmill-trained dogs and found reduced temporal parameters during backward walking and a marked reduction in wrist palmar-flexion during the swing phase of a backward step, but no change in trunk posture. We suggest that the aberrant posture exhibited by cats during backward walking is more related to ethological factors than to biomechanical ones. PMID- 10828321 TI - Effects of speed and distance of muscle shortening on force depression during voluntary contractions. AB - The purpose of this study was to determine the influence of speed and distance of muscle shortening on the amount of force depression for voluntary contractions. Two experimental tests were performed. In the first test, subjects performed isometric knee extensor contractions following muscle shortening produced by isokinetic knee extensions over the range 25-50 degrees. In the second test, subjects performed isometric knee extensor contractions following muscle shortening produced by isokinetic knee extensions at two speeds: 20 and 240 degrees /s. Knee extensor moments, surface electromyographical (EMG) signals of quadriceps femoris, and interpolated twitch moments were measured during all contractions and were compared with the corresponding values obtained during purely isometric contractions. Force depression following muscle shortening for the voluntary contractions tested in this study did not depend on the distance or the speed of muscle shortening. These results are in contrast to the corresponding results in the literature obtained using artificial electrical stimulation in which force depression was always found to be directly related to the distance of shortening and inversely related to the speed of shortening. The difference in force depression as a function of the distance and speed of muscle shortening between voluntary and artificial electrical stimulation may be associated with changes in activation following the voluntary shortening contractions, whereas activation is controlled and constant in all artificial stimulation protocols. PMID- 10828322 TI - Simplified dynamics model of planar two-joint arm movements. AB - A theoretical framework is presented that describes a way in which the inverse dynamics equations of motion of planar two-joint arm movements (EX-model) are reformulated in a simple form. A single point was assumed to define both the wrist and elbow joint centers, and thus the motion of two points in extrinsic space was represented by second-order differential equations to provide the variables in the reformulation (RE-) model. Through an analytical processes, it was shown that the RE-model for reproducing the shoulder joint torque consists of the linearly scaled moment per unit mass responsible for accelerating the wrist and elbow points about the shoulder joint, while that for reproducing the elbow joint torque consists of the linearly scaled moment per unit mass responsible for accelerating the wrist point about the elbow. The scaling factors for variables in the RE-model were based solely on the values for segment lengths, while in the EX-model the inertial parameter data for the segments are involved in its representation. The inertial parameter data of six-arm specimens from the cadaver experiment of Chandler et al. (1975, AMRL Technical Report, Wright-Patterson Air Force Base, OH) were used to develop and verify the numeric solutions of the RE model. The adequacy of the model varied somewhat among subjects, but minor changes of the physical parameters of the arm segments enabled perfect reformulation, regardless of the specimens. The potential abilities of the RE model to deal with the complexities in motor control with more simple control schemes are discussed. PMID- 10828323 TI - Non-linearities in apparent mass and transmissibility during exposure to whole body vertical vibration. AB - The causes of low back pain associated with prolonged exposure to whole-body vibration are not understood. An understanding of non-linearities in the biomechanical responses is required to identify the mechanisms responsible for the dynamic characteristics of the body, to allow for the non-linearities when predicting the influence of seating dynamics, and to predict the adverse effects caused by various magnitudes of vibration. Twelve subjects were exposed to six magnitudes, 0.25-2.5ms(-2) rms, of vertical random vibration in the frequency range 0.2-20Hz. The apparent masses of the subjects were determined together with transmissibilities measured from the seat to various locations on the body surface: the upper and lower abdominal wall, at L3, over the posterior superior iliac spine and the iliac crest. There were significant reductions in resonance frequencies for both the apparent mass and the transmissibilities to the lower abdomen with increases in vibration magnitude. The apparent mass resonance frequency reduced from 5.4-4. 2Hz as the magnitude of the vibration increased from 0.25-2.5ms(-2) rms. Vertical motion of the lumbar spine and pelvis showed resonances at about 4Hz and between 8 and 10Hz. When exposed to vertical vibration, the human body shows appreciable non-linearities in its biodynamic responses. Biodynamic models should be developed to reflect the non-linearity. PMID- 10828324 TI - The isometric functional capacity of muscles that cross the elbow. AB - We hypothesized that muscles crossing the elbow have fundamental differences in their capacity for excursion, force generation, and moment generation due to differences in their architecture, moment arm, and the combination of their architecture and moment arm. Muscle fascicle length, sarcomere length, pennation angle, mass, and tendon displacement with elbow flexion were measured for the major elbow muscles in 10 upper extremity specimens. Optimal fascicle length, physiological cross-sectional area (PCSA), moment arm, operating range on the force-length curve, and moment-generating capacity were estimated from these data. Brachioradialis and pronator teres had the longest (17.7cm) and shortest (5.5cm) fascicles, respectively. Triceps brachii (combined heads) and brachioradialis had the greatest (14.9cm(2)) and smallest (1.2cm(2)) PCSAs, respectively. Despite a comparable fascicle length, long head of biceps brachii operates over a broader range of the force-length curve (length change=56% of optimal length, 12.8cm) than the long head of triceps brachii (length change=28% of optimal length, 12. 7cm) because of its larger moment arm (4.7cm vs. 2.3cm). Although brachioradialis has a small PCSA, it has a relatively large moment generating capacity (6.8cm(3)) due to its large moment arm (average peak=7.7cm). These results emphasize the need to consider the interplay of architecture and moment arm when evaluating the functional capabilities of a muscle. PMID- 10828325 TI - The influence of the acetabular labrum on hip joint cartilage consolidation: a poroelastic finite element model. AB - The goal of this study was to investigate the influence of the acetabular labrum on the consolidation, and hence the solid matrix strains and stresses, of the cartilage layers of the hip joint. A plane-strain finite element model was developed, which represented a coronal slice through the acetabular and femoral cartilage layers and the acetabular labrum. Elements with poroelastic properties were used to account for the biphasic solid/fluid nature of the cartilage and labrum. The response of the joint over an extended period of loading (10,000s) was examined to simulate the nominal compressive load that the joint is subjected to throughout the day. The model demonstrated that the labrum adds an important resistance in the flow path of the fluid being expressed from the cartilage layers of the joint. Cartilage layer consolidation was up to 40% quicker in the absence of the labrum. Following removal of the labrum from the model, the solid on-solid contact stresses between the femoral and acetabular cartilage layers were greatly increased (up to 92% higher), which would increase the friction between the joint surfaces. In the absence of the labrum, the centre of contact shifted towards the acetabular rim. Subsurface strains and stresses were much higher without the labrum, which could contribute to fatigue damage of the cartilage layers. Finally, the labrum provided some structural resistance to lateral motion of the femoral head within the acetabulum, enhancing joint stability and preserving joint congruity. PMID- 10828326 TI - A fuzzy logic model of fracture healing. AB - A quantitative biomechanical model describes the tissue transformation during healing of a transverse osteotomy of a sheep metatarsal. The model predicts bridging of the bone ends through cartilage, followed by the growth of a callus cuff, and finally, the resorption of callus after ossification of the interfragmentary gap. We suggest bone density or the modulus of elasticity do not sufficiently characterize healing tissue for predictive purposes. In addition to the stimulus reflected by strain energy density we introduce a new osteogenic factor based upon stress gradients and which predicts areas of a high osteogenic capacity. Our model distinguishes three basic types of tissue, namely bone, cartilage and fibrous tissue. A fuzzy controller is proposed to model the tissue reaction. A set of fuzzy rules derived from medical knowledge has been implemented to describe tissue transformation such as intramembraneous or chondral ossification, atrophy or destruction. Fuzzy logic is able to model tissue transformation processes within the numerical simulation of remodeling processes. This approach improves the simulation tools and affords the potential to optimize planning of animal experiments and conduct parametric studies. PMID- 10828327 TI - A governing relationship for repetitive muscular contraction. AB - During repetitive contractions, muscular work has been shown to exhibit complex relationships with muscle strain length, cycle frequency, and muscle shortening velocity. Those complex relationships make it difficult to predict muscular performance for any specific set of movement parameters. We hypothesized that the relationship of impulse with cyclic velocity (the product of shortening velocity and cycle frequency) would be independent of strain length and that impulse cyclic velocity relationships for maximal cycling would be similar to those of in situ muscle performing repetitive contraction. Impulse and power were measured during maximal cycle ergometry with five cycle-crank lengths (120-220mm). Kinematic data were recorded to determine the relationship of pedal speed with joint angular velocity. Previously reported in situ data for rat plantaris were used to calculate values for impulse and cyclic velocity. Kinematic data indicated that pedal speed was highly correlated with joint angular velocity at the hip, knee, and ankle and was, therefore, considered a valid indicator of muscle shortening velocity. Cycling impulse-cyclic velocity relationships for each crank length were closely approximated by a rectangular hyperbola. Data for all crank lengths were also closely approximated by a single hyperbola, however, impulse produced on the 120mm cranks differed significantly from that on all other cranks. In situ impulse-cyclic velocity relationships exhibited similar characteristics to those of cycling. The convergence of the impulse-cyclic velocity relationships from most crank and strain lengths suggests that impulse cyclic velocity represents a governing relationship for repetitive muscular contraction and thus a single equation can predict muscle performance for a wide range of functional activities. The similarity of characteristics exhibited by cycling and in situ muscle suggests that cycling can serve as a window though which to observe basic muscle function and that investigators can examine similar questions with in vivo and in situ models. PMID- 10828328 TI - Blood flow and vessel mechanics in a physiologically realistic model of a human carotid arterial bifurcation. AB - The pulsatile flow in an anatomically realistic compliant human carotid bifurcation was simulated numerically. Pressure and mass flow waveforms in the carotid arteries were obtained from an individual subject using non-invasive techniques. The geometry of the computational model was reconstructed from magnetic resonance angiograms. Maps of time-average wall shear stress, contours of velocity in the flow field as well as wall movement and tensile stress on the arterial wall are all presented. Inconsistent with previous findings from idealised geometry models, flow in the carotid sinus is dominated by a strong helical flow accompanied by a single secondary vortex motion. This type of flow is induced primarily by the asymmetry and curvature of the in vivo geometry. Flow simulations have been carried out under the rigid wall assumption and for the compliant wall, respectively. Comparison of the results demonstrates the quantitative influence of the vessel wall motion. Generally there is a reduction in the magnitude of wall shear stress, with its degree depending on location and phase of the cardiac cycle. The region of slow or reversed flow was greater, in both spatial and temporal terms in the compliant model, but the global characteristics of the flow and stress patterns remain unchanged. The analysis of mechanical stresses on the vessel surface shows a complicated stress field. Stress concentration occurs at both the anterior and posterior aspects of the proximal internal bulb. These are also regions of low wall shear stress. The comparison of computed and measured wall movement generally shows good agreement. PMID- 10828329 TI - Kinematics of human arm reconstructed from spatial tracking system recordings. AB - The kinematics of the human arm in terms of angles of rotations in the joints is reconstructed from the spatial tracking system (Fastrack() Polhemus) recordings. The human arm is modeled by three rigid bodies (the upper arm, the forearm and the hand) with seven degrees of freedom (three in the shoulder, two in the elbow and two in the wrist). Joint geometry parameters (orientations of the axes relative to the arm segments, the angles and the distances between the axes) have been calculated on the basis of passive rotations in the joints. The calculated parameters have been used to solve the direct kinematics problem for the reaching movements in different directions. The difference between calculated and recorded positions and accelerations of the hand has been used to assess the accuracy of the proposed method of kinematics reconstruction. The error analysis showed that spatial tracking system recordings and human arm kinematics reconstruction could reliably be used to accurately analyze multijoint movement in humans. PMID- 10828330 TI - Investigation of the kinetics of anaerobic metabolism by analysis of the performance of elite sprinters. AB - The principal motivation for the present work was the study of the kinetics of anaerobic metabolism. A new mathematical model of the bioenergetics of sprinting, incorporating a three-equation representation of anaerobic metabolism, is developed. Results computed using the model are compared with measured data from the mens' finals of the 100m event at the 1987 World Championships. The computed results closely predict the overall average performance of the competitors over the course of the entire race. Further calculations show the three-equation model of anaerobic metabolism to be a significant improvement over the previous one equation model. Representative values of time constants that govern the rate of anaerobic energy release have been determined for elite male athletes. For phosphocreatine utilisation, values for lambda(2)=0. 20s(-1) and psi(2)=3.0s(-1) are consistent with data previously reported in the literature. New values of lambda(3)=0.033s(-1) and psi(3)=0.34s(-1) are proposed as offering an improved representation of the kinetics of oxygen-independent glycolysis. For the first time, tentative values for the time constants of ATP utilisation, lambda(1)=0.9s( 1) and psi(1)=20s(-1), are suggested. The maximum powers developed during sprinting by oxygen-independent glycolysis, PCr utilisation and endogenous ATP utilisation were calculated as 34. 1, 30.1 and 16.6Wkg(-1), respectively, with an overall maximum anaerobic power of 51.6Wkg(-1). Sample calculations show the mathematical model can be used in principle to derive data on the kinetics of anaerobic metabolism of individual athletes. PMID- 10828331 TI - The mesh-matching algorithm: an automatic 3D mesh generator for finite element structures. AB - Several authors have employed finite element analysis for stress and strain analysis in orthopaedic biomechanics. Unfortunately, the definition of three dimensional models is time consuming (mainly because of the manual 3D meshing process) and consequently the number of analyses to be performed is limited. The authors have investigated a new patient-specific method allowing automatically 3D mesh generation for structures as complex as bone for example. This method, called the mesh-matching (M-M) algorithm, generated automatically customized 3D meshes of anatomical structures from an already existing model. The M-M algorithm has been used to generate FE models of 10 proximal human femora from an initial one which had been experimentally validated. The automatically generated meshes seemed to demonstrate satisfying results. PMID- 10828332 TI - Three-dimensional numerical simulations of flow through a stenosed coronary bypass. AB - This work analyzes the flow patterns at the anastomosis of a stenosed coronary bypass. Three-dimensional numerical simulations are performed using a finite elements method. We consider a geometrical model of the host coronary artery with and without a 75% severity stenosis for three different locations from the anastomosis. The flow features - velocity profiles, secondary motions and wall shear stresses - are compared for different configurations of the flow rate and of the distance of the anastomosis from the site of occlusion (called distance of grafting). The combination of the junction flow effects - counter rotating vortices - with the stenosis effects - confined jet flow - is particularly important when the distance of grafting is short. Given that the residual flow issued from the pathologic stenosis being non-negligible after two weeks grafting, models without stenosis cannot predict the evolution of the wall shear stress in the vicinity of the anastomosis. PMID- 10828333 TI - The accuracy of joint surface models constructed from data obtained with an electromagnetic tracking device. AB - Electromagnetic tracking devices are widely used in biomechanics. In this article a method is evaluated to construct models of articular surfaces using an electromagnetic tracking device. First, the accuracy of the space tracker was examined and optimised. Then, from several joint surfaces random points were measured and eighth degree polynomials were fitted to these measurements. To check if the fit converged well, plots of cross sections of the model with corresponding data points were examined. The accuracy of the models was determined by comparing them with computed tomography data and by reproducibility tests. All the fits converged well to the data. The root mean square (RMS) error of the models varied from 0.07 to 0. 18mm, and was proportional to the size and complexity of the surface. This was mainly due to systematic errors made by the space tracker, which were also proportional to the size and complexity of the surface. PMID- 10828334 TI - Measurement of the screw-home motion of the knee is sensitive to errors in axis alignment. AB - Measurements of joint angles during motion analysis are subject to error caused by kinematic crosstalk, that is, one joint rotation (e. g., flexion) being interpreted as another (e.g., abduction). Kinematic crosstalk results from the chosen joint coordinate system being misaligned with the axes about which rotations are assumed to occur. The aim of this paper is to demonstrate that measurement of the so-called "screw-home" motion of the human knee, in which axial rotation and extension are coupled, is especially prone to errors due to crosstalk. The motions of two different two-segment mechanical linkages were examined to study the effects of crosstalk. The segments of the first linkage (NSH) were connected by a revolute joint, but the second linkage (SH) incorporated gearing that caused 15 degrees of screw-home rotation to occur with 90 degrees knee flexion. It was found that rotating the flexion axis (inducing crosstalk) could make linkage NSH appear to exhibit a screw-home motion and that a different rotation of the flexion axis could make linkage SH apparently exhibit pure flexion. These findings suggest that the measurement of screw-home rotation may be strongly influenced by errors in the location of the flexion axis. The magnitudes of these displacements of the flexion axis were consistent with the inter-observer variability seen when five experienced observers defined the flexion axis by palpating the medial and lateral femoral epicondyles. Care should be taken when interpreting small internal-external rotations and abduction adduction angles to ensure that they are not the products of kinematic crosstalk. PMID- 10828335 TI - Force output during and following active stretches of rat plantar flexor muscles: effect of velocity of ankle rotation. AB - During the development of force deficits by repeated stretches, velocity sensitive changes in the extra force produced during and after subsequent stretching has not been studied. In the present study, repeated dorsiflexion of the foot of rats with maximally contracting plantar flexor muscles was performed at two angular velocities [0.87 (slow muscle stretch) and 10.47rads(-1) (fast muscle stretch)] to examine the active force of the muscles during and following dorsiflexion. Dorsiflexion was performed 30 times with a rest period of 3min between the stretches to minimize muscle fatigue. The ability of rat plantar flexor muscles to produce additional force during the stretch was not velocity sensitive. In contrast, repeated dorsiflexion with fast muscle stretches, but not with slow muscle stretches, resulted in an increase in the force decay with time following the stretches (i.e. increased stress relaxation), as indicated by a change in the time constant of force decay during stress relaxation. Apparently, the stress-relaxation of rat plantar flexor muscles is sensitive to angular velocity of ankle movements; repeated fast, but not slow dorsiflexion, alters the stress relaxation process of active skeletal muscles exposed to stretches which create a force deficit. The change in time constant of force decay during stress relaxation in response to a series of repeated stretches might provide information on the sarcomere length distribution in skeletal muscles. PMID- 10828336 TI - Calibration of position and angular data from a magnetic tracking device. AB - This paper describes a method for calibrating data from a magnetic tracking device. Position and orientation data were collected in a 1. 6x0.8x1.4m(3) volume using a Polhemus Fastrak((R)) in conjunction with both a long-range and standard transmitter. Position and orientation data were calibrated using a locally linear model based on the position of the measurement. After calibration, the average position and angular errors were less than 1.8cm and 1.2 degrees up to 1.8m from the transmitter for the long-range transmitter. For the standard transmitter, even after calibration, errors increased sharply when the sensor was more than 1.2m from the transmitter. Up to that distance, post-calibration errors were less than 1.2cm and 1. 2 degrees, while up to 1.8m they were below 5cm and 4 degrees. These errors could be further reduced by noise filtering. However, use of the standard transmitter is not recommended at distance greater than 1.2m due to orientation-based effects. It was concluded that for the volume investigated, tracking devices could provide similar three-dimensional accuracy to video systems. PMID- 10828337 TI - Roles of periaqueductal gray and nucleus tractus solitarius in cardiorespiratory function in the rat brainstem. AB - Periaqueductal gray (PAG) and nucleus tractus solitarius (NTS) are important centres for regulation of cardiorespiratory function in cats. We aimed to study the effects of specific PAG stimulation on cardiorespiratory parameters in the rat. Microinjection of D, L-homocysteic acid (DLH) into dorsolateral PAG of anaesthetised rats, led to: marked increases in respiratory frequency (RF) and amplitude of diaphragmatic electromyogram, decreases in inspiratory and expiratory durations, and increased blood pressure and heart rate. Following injection of propranolol (150 pmol, 30 nl), a beta-adrenergic antagonist, into the commissural subnucleus of NTS, the DLH-induced increase in RF was markedly attenuated. Inspiratory neurones (late I cells) in NTS were excited upon stimulation of PAG and their increased activity was accompanied by increased RF. The changes in activity of the late I cells in response to stimulation of dorsolateral PAG provide physiological evidence of a link, possibly noradrenergic, between the two nuclei and involvement of the NTS in control of respiratory functions orchestrated by the PAG. PMID- 10828338 TI - The influence of descending inputs on breathing pattern formation in the isolated bullfrog brainstem-spinal cord. AB - This study used in vitro brainstem-spinal cord preparations from the American bullfrog, Rana catesbeiana, to examine the influence of central descending inputs on breathing pattern formation. In preparations with an episodic pattern of fictive breathing, a transection slightly caudal to the optic chiasma produced a continuous breathing pattern and increased the overall frequency of fictive breathing. Following a transection to isolate the medulla, the frequency of fictive breathing decreased and the incidence of other forms of motor output increased. Further transections between the trigeminal and vagus nerve roots resulted in variable and asynchronous discharge from each nerve. These results suggest that a primary respiratory rhythm is produced within the medulla but descending influences stimulate breathing and promote episodic breathing. It would appear that multiple elements of the respiratory control system, including tegmental and medullary sites, play a role in shaping the burst pattern of motor output associated with each breath and that slower rhythms of longer burst duration are generated by more caudal hindbrain sites. PMID- 10828339 TI - Pulmonary vagal modulation of ventilation in toads (Bufo marinus). AB - This study examined the role of pulmonary vagal feedback on hypercapnic chemosensitivity and breathing pattern formation in cane toads (Bufo marinus). Decerebrate, paralysed toads were uni-directionally ventilated with air, 2.5% CO(2) or 5.0% CO(2) with the lungs inflated or deflated, before and after pulmonary vagotomy. Motor output from the mandibular branch of the trigeminal nerve served as an index of fictive breathing. As respiratory drive was increased, breathing frequency increased and breaths were clustered into discrete episodes separated by periods of apnea. Lung deflation tended to enhance episodic breathing while inflation biased the system towards apnea at low levels of respiratory drive and a pattern of continuous, small breaths at higher levels of respiratory drive. Following bilateral pulmonary vagotomy there was no increase in ventilation during hypercapnia and lung inflation/deflation had no effect on breathing pattern. In isolated brainstem-spinal cord preparations from the same animals, all variables associated with fictive breathing were unaffected by changes in superfusate pH from 8.0 to 7.6. The breathing pattern from the in vitro preparations was highly variable. This study demonstrates a crucial role for vagal feedback in modulating respiration and the respiratory responses to hypercapnia in B. marinus. PMID- 10828340 TI - Effect of alveolar volume and sequential filling on the diffusing capacity of the lungs: I. theory. AB - The diffusing capacity, DL, is a critical physiological parameter of the lung used to assess gas exchange clinically. Most models developed to analyze experimental data from a single breath maneuver have assumed a well-mixed or uniform alveolar region, including the clinically accepted Jones-Meade method. In addition, all previous models have assumed a constant DL, which is independent of alveolar volume, VA. In contrast, experimental data provide evidence for a non uniform alveolar region coupled with sequential filling of the lung. In addition, although the DL for carbon monoxide is a weak function of VA, the DL of nitric oxide depends strongly on VA. We have developed a new mathematical model of the single breath maneuver that considers both a variable degree of sequential filling and a variable DL. Our model predicts that the Jones-Meade method overestimates DL when the exhaled gas sample is collected late in the exhalation, but underestimates DL if the exhaled gas sample is collected early in the exhalation phase due to the effect of sequential filling. Utilizing a prolonged constant exhalation method, or a three-equation method, will also produce erroneous predictions of DL. We conclude that current methods may introduce significant error in the estimation of DL by ignoring the sequential filling of the lung, and the dependence of DL on VA. PMID- 10828341 TI - Effect of alveolar volume and sequential filling on the diffusing capacity of the lungs: II. Experiment. AB - The diffusing capacity of the lung, DL, is a critical physiological parameter, yet the currently accepted clinical model (Jones-Meade) assumes a well-mixed alveolar region, and a constant DL independent of alveolar volume, VA, despite experimental evidence to the contrary. We have formulated a new mathematical model [Tsoukias, N.M, Wilson, A.F., George, S.C., 2000. Respir. Physiol. 120, 231 249] that considers variable alveolar mixing through a single parameter, k (05 or 80% of patients throughout the 40-week study. At weeks 2, 8, 24, and 40, disease severity was 'much improved' or 'very much improved' in 49, 58, 59, and 58% of patients, respectively. The mean (+/-SEM) ESS score improved significantly from 16.5+/-0.2 at open-label baseline to 12.4+/-0.2 at week 2 and remained at that level through week 40 (P<0.001). Quality of life scores at weeks 4, 8, 24, and 40 were significantly improved versus open-label baseline scores for six of the eight SF-36 domains (P<0.001). The most common treatment-related adverse events were headache (13%), nervousness (8%), and nausea (5%). Most adverse events were mild to moderate in nature. A total of 341 patients (71%) completed the studies. Forty-three patients (9.0%) discontinued treatment because of adverse events.Conclusions: Modafinil is effective for the long-term treatment of EDS associated with narcolepsy and significantly improves perceptions of general health. Modafinil is well tolerated, with no evidence of tolerance developing during 40 weeks of treatment. PMID- 10828435 TI - Journal search and commentary. PMID- 10828436 TI - Articles reviewed: 1. Sleep deprivation-induced reduction in cortical functional response to serial subtraction. 2. Altered brain response to verbal learning following sleep deprivation. PMID- 10828437 TI - Article reviewed: Reduction of rapid eye movement sleep by diurnal and nocturnal seizures in temporal lobe epilepsy. PMID- 10828438 TI - Article reviewed: Sleep apnea and daytime sleepiness and fatigue: related to visceral obesity, insulin resistance, and hypercytokinemia. PMID- 10828439 TI - Article reviewed: The novel brain neuropeptide, orexin-A, modulates the sleep wake cycle of rats. PMID- 10828440 TI - Mitochondrial DNA depletion causes morphological changes in the mitochondrial reticulum of cultured human cells. AB - Depletion of mitochondrial DNA (mtDNA) causes defects in respiratory activity and energy production. Recent studies have shown mitochondria to exist primarily as reticular networks, having tubular cristae. Using fluorescence microscopy and transmission electron microscopy, we have examined mitochondrial morphology and interior structure in wildtype and mtDNA-depleted rho0 human fibroblasts and 143B osteosarcoma cell lines. MtDNA depletion results in compromise of the mitochondrial continuum and causes a reduction in amount of cristal membranes, often prompting the remaining cristae to adopt a circular appearance in the mitochondrial interior. These changes emphasize the tight relationship between mitochondrial structure and function. PMID- 10828441 TI - Cloning and expression analysis of an inducible HSP70 gene from tilapia fish. AB - We isolated and characterized the tilapia (Oreochromis mossambicus) HSP70 gene, highly homologous to other HSP70 genes. A dramatic increase of tilapia HSP70 mRNA levels was observed after heat shock of whole animals in all organs tested. Reporter constructs were tested for transient expression in carp cells and in microinjected zebrafish embryos. The entire isolated regulatory region ( 851/+157) was able to mediate heat shock inducible expression of the reporter gene, with no preference for a particular tissue. Our studies represent the first transcriptional analysis of a HSP70 promoter from fish, revealing a powerful tool to direct controlled, tissue-independent gene expression in fish. PMID- 10828442 TI - Involvement of poly(ADP-ribose) polymerase and activation of caspase-3-like protease in heat shock-induced apoptosis in tobacco suspension cells. AB - The cleavage of poly(ADP-ribose) polymerase (PARP) by caspase (casp)-3 is an essential link in the apoptotic pathway in animal cells. In plant cells, however, there is no authentic evidence for the similar role that PARP may play during apoptosis. Using a heat shock (HS)-induced apoptosis system of tobacco cells, we found that immediately after a 4 h heat treatment, PARP was cleaved to form an 89 kDa signature fragment, while DNA laddering appeared only after a 20 h recovery following the HS. An activation of casp-3-like protease was also observed. The results suggest that apoptosis in plants and animals may share common mechanisms. On the other hand, when cells were preincubated with 4 mM 3-aminobenzamide or 2-8 mM nicotinamide, the specific inhibitors of PARP, before HS treatment, apoptotic cell death was reduced significantly. Our results thus imply that PARP may also be involved in apoptosis in a different way from the casp-related events. PMID- 10828443 TI - Novel Dictyostelium unconventional myosin, MyoM, has a putative RhoGEF domain. AB - We have cloned a novel unconventional myosin gene myoM in Dictyostelium. Phylogenetic analysis of the motor domain indicated that MyoM does not belong to any known subclass of the myosin superfamily. Following the motor domain, two calmodulin-binding IQ motifs, a putative coiled-coil region, and a Pro, Ser and Thr-rich domain, lies a combination of dbl homology and pleckstrin homology domains. These are conserved in Rho GDP/GTP exchange factors (RhoGEFs). We have identified for the first time the RhoGEF domain in the myosin sequences. The growth and terminal developmental phenotype of Dictyostelium cells were not affected by the myoM(-) mutation. Green fluorescent protein-tagged MyoM, however, accumulated at crown-shaped projections and membranes of phase lucent vesicles in growing cells, suggesting its possible roles in macropinocytosis. PMID- 10828444 TI - Multiprotein complexes present at the MIF motifs flanking the promoter of the human c-myc gene. AB - The activated c-myc allele in Burkitt's lymphoma is associated with a clustering of somatic mutations within a discrete domain of intron I that define protein recognition sequences, designated as myc intron factors (MIF-1, MIF-2 and MIF-3). We have previously shown that MIF-1 binding activity consists of two polypeptides, myc intron binding polypeptide (MIBP1) and RFX1. In the present study we identified two polypeptides, p105 and p115, and showed that these proteins give rise to a DNA-protein complex at the MIF-2 as well as the adjacent MIF-1 site. In addition, we demonstrated that all four proteins interact with a novel MIF-1 like motif upstream from the c-myc promoter region, designated 5'MIF. These data suggest a model, where the interactions of MIBP1/RFX1 and p105/p115 with the MIF-like sites may play a role in the promoter topology of the c-myc gene. PMID- 10828445 TI - The effect of ethylene on MAPKinase-like activity in Arabidopsis thaliana. AB - Protein kinase activity was studied in cytosolic extracts from leaves of wild type Arabidopsis thaliana, the ethylene-insensitive mutant, etr1, and the constitutive triple-response mutant, ctr1. Treatment of wild type with ethylene resulted in increased myelin basic protein (MBP) phosphorylation. In etr1, constitutive protein kinase activity was lower than in wild type, but in ctr1, activity was enhanced. A protein of M(r) approximately 47 kDa associated with MBP phosphorylating activity was detected using in gel protein kinase assays and phosphorylation of this protein was promoted by ethylene treatment in wild type while activity in the mutants reflected that of MBP phosphorylation. Both MAPKinase (ERK 1) and phosphotyrosine antibodies immunoprecipitated MBP phosphorylating activity and detected a polypeptide band at M(r) approximately 47 kDa. Immunoprecipitated MBP-phosphorylating activity was again much lower in etr1 compared to wild type but much higher in ctr1. Antibodies to phosphorylated MAPKinase recognised proteins at approximately 47 kDa and the signal was upregulated in response to ethylene. The data obtained suggest that the detected protein(s) is a MAPKinase and provide further evidence confirming that a MAPKinase cascade(s) is involved in ethylene signal transduction. PMID- 10828446 TI - Identification of key amino acid residues in Neisseria polysaccharea amylosucrase. AB - Amylosucrase from Neisseria polysaccharea catalyzes the synthesis of an amylose like polymer from sucrose. Sequence alignment revealed that it belongs to the glycoside hydrolase family 13. Site-directed mutagenesis enabled the identification of functionally important amino acid residues located at the active center. Asp-294 is proposed to act as the catalytic nucleophile and Glu 336 as general acid base catalyst in amylosucrase. The conserved Asp-401, His-195 and His-400 residues are critical for the enzymatic activity. These results provide strong support for the predicted close structural and functional relationship between the sucrose-glucosyltransferases and enzymes of the alpha amylase family. PMID- 10828447 TI - Cytoplasmic localization of the interferon-inducible protein that is encoded by the AIM2 (absent in melanoma) gene from the 200-gene family. AB - While interferons (IFNs) (alpha, beta and gamma), a family of cytokines, have the ability to exert the growth-inhibitory effect on target cells, the molecular mechanism(s) by which IFNs inhibit cell growth remains to be identified. Because IFN-inducible 'effector' proteins mediate the biological activities of IFNs, characterization of IFN-inducible proteins is critical to identify their functional role in IFN action. One family (the 200-family) of IFN-inducible proteins is encoded by structurally related murine (Ifi202a, Ifi202b, Ifi203, Ifi204 and D3) and human (IFI16, MNDA and AIM2) genes. The proteins encoded by genes in the family share a unique repeat of 200-amino acids and are primarily nuclear. The AIM2 gene is a newly identified gene that is not expressed in a human melanoma cell line. Here we report that AIM2 is estimated to be a 39 kDa protein and, unlike other proteins in the family, is localized primarily in the cytoplasm. Interestingly, overexpression of AIM2 in transfected cells retards proliferation and, under reduced serum conditions, increases the susceptibility to cell death. Moreover, AIM2 can heterodimerize with p202 in vitro. Together, these observations provide support to the idea that AIM2 may be an important mediator of IFN action. PMID- 10828448 TI - Abscissic acid specific expression of RAB18 involves activation of anion channels in Arabidopsis thaliana suspension cells. AB - The abscissic acid (ABA) transduction cascade following the plasmalemma perception was analyzed in intact Arabidopsis thaliana suspension cells. In response to impermeant ABA, anion currents were activated and K(+) inward rectifying currents were inhibited. Anion current activation was required for the ABA specific expression of RAB18. By contrast, specific inhibition of K(+) channels by tetraethylammonium or Ba(2+) did not affect RAB18 expression. Thus, outer plasmalemma ABA perception triggered two separated signaling pathways. PMID- 10828449 TI - 14-3-3 proteins interact with a 13-lipoxygenase, but not with a 9-lipoxygenase. AB - Associations between lipoxygenases (Lox) and 14-3-3 proteins were demonstrated by two different methods. First, immunoprecipitation experiments, using isoenzyme specific monoclonal Lox antibodies, showed that 14-3-3 proteins co-precipitate with 13-Lox, but not with the 9-Lox from barley. Second, interactions between 13 Lox and 14-3-3 were established by surface plasmon resonance studies, showing that 13-Lox binds with 14-3-3 proteins in a concentration-dependent manner. The interactions between 14-3-3 proteins and 13-Lox may reveal their role during plant development. PMID- 10828450 TI - The role of mild uncoupling and non-coupled respiration in the regulation of hydrogen peroxide generation by plant mitochondria. AB - The roles of mild uncoupling caused by free fatty acids (mediated by plant uncoupling mitochondrial protein (PUMP) and ATP/ADP carrier (AAC)) and non coupled respiration (alternative oxidase (AO)) on H(2)O(2) formation by plant mitochondria were examined. Both laurate and oleate prevent H(2)O(2) formation dependent on the oxidation of succinate. Conversely, these free fatty acids (FFA) only slightly affect that dependent on malate plus glutamate oxidation. Carboxyatractylate (CAtr), an inhibitor of AAC, completely inhibits oleate- or laurate-stimulated oxygen consumption linked to succinate oxidation, while GDP, an inhibitor of PUMP, caused only a 30% inhibition. In agreement, CAtr completely restores the oleate-inhibited H(2)O(2) formation, while GDP induces only a 30% restoration. Both oleate and laurate cause a mild uncoupling of the electrical potential (generated by succinate), which is then followed by a complete collapse with a sigmoidal kinetic. FFA also inhibit the succinate-dependent reverse electron transfer. Diamide, an inhibitor of AO, favors the malate plus glutamate dependent H(2)O(2) formation, while pyruvate (a stimulator of AO) inhibits it. These results show that the succinate-dependent H(2)O(2) formation occurs at the level of Complex I by a reverse electron transport. This generation appears to be prevented by mild uncoupling mediated by FFA. The anionic form of FFA appears to be shuttled by AAC rather than PUMP. The malate plus glutamate-dependent H(2)O(2) formation is, conversely, mainly prevented by non-coupled respiration (AO). PMID- 10828451 TI - Novel intracellular effects of human connective tissue growth factor expressed in Cos-7 cells. AB - To clarify the multiple functionality of connective tissue growth factor (CTGF), we examined the effects of nascent CTGF within the cell by transient expression. In Cos-7 cells, expression of human CTGF induced an altered cell morphology. It was associated with an increased cellular DNA content and loose attachment, indicating the cells were in G2/M phase. Overexpression of CTGF did not induce cell growth, whereas recombinant CTGF efficiently stimulated the proliferation extracellularly. These results indicate that intracellular CTGF may act as an antimitotic agent, thus it should also be noted that nascent CTGF was found to accumulate around the central mitotic machinery. PMID- 10828452 TI - D-Serine inhibits serine palmitoyltransferase, the enzyme catalyzing the initial step of sphingolipid biosynthesis. AB - Serine palmitoyltransferase (SPT), responsible for the initial step of sphingolipid biosynthesis, catalyzes condensation of palmitoyl coenzyme A and L serine to produce 3-ketodihydrosphingosine (KDS). For determination of the stereochemical specificity of the amino acid substrate, a competition analysis of the production of [(3)H]KDS from L-[(3)H]serine was performed using purified SPT. D-Serine inhibited [(3)H]KDS production as effectively as non-radioactive L serine, whereas neither D-alanine nor D-threonine showed any significant effect. Incubation of purified SPT with [palmitoyl 1-(14)C]palmitoyl coenzyme A and D serine did not produce [(14)C]KDS, while the control incubation with L-serine did. These results suggest that D-serine competes with L-serine for the amino acid recognition site of SPT, but that D-serine is not utilized by this enzyme to produce KDS. PMID- 10828453 TI - The Rab3-interacting molecule RIM is expressed in pancreatic beta-cells and is implicated in insulin exocytosis. AB - The putative Rab3 effector RIM (Rab3-interacting molecule) was detected by Northern blotting, RT-PCR and Western blotting in native pancreatic beta-cells as well as in the derived cell lines INS-1E and HIT-T15. RIM was localized on the plasma membrane of INS-1E cells and beta-cells. An involvement of RIM in insulin exocytosis was indicated by transfection experiments of INS-1E cells with the Rab3 binding domain of RIM. This domain enhanced glucose-stimulated secretion in intact cells and Ca(2+)-stimulated exocytosis in permeabilized cells. Co expression of Rab3A reversed the effect of RIM on exocytosis. These results suggest an implication of RIM in the control of insulin secretion. PMID- 10828454 TI - Dominant negative myostatin produces hypertrophy without hyperplasia in muscle. AB - Myostatin, a TGF-beta family member, is a negative regulator of muscle growth. Here, we generated transgenic mice that expressed myostatin mutated at its cleavage site under the control of a muscle specific promoter creating a dominant negative myostatin. These mice exhibited a significant (20-35%) increase in muscle mass that resulted from myofiber hypertrophy and not from myofiber hyperplasia. We also evaluated the role of myostatin in muscle degenerative states, such as muscular dystrophy, and found significant downregulation of myostatin. Thus, further inhibition of myostatin may permit increased muscle growth in muscle degenerative disorders. PMID- 10828455 TI - The primary structure of the acidic lectin from winged bean (Psophocarpus tetragonolobus): insights in carbohydrate recognition, adenine binding and quaternary association. AB - The amino acid sequence of the winged bean acidic lectin (WBA II) was determined by chemical means and by recombinant techniques. From the N- and C-terminal sequence, obtained chemically, primers were designed for PCR amplification of the genomic DNA. The PCR product was cloned and sequenced to get the complete primary structure of WBA II. Peptide fragments for sequencing were also obtained by tryptic cleavages of the native lectin. The WBA II sequence showed a high degree of homology with that of WBA I and Erythrina corallodendron lectin (ECorL), especially in the regions involved in subunit association, where there is a very high conservation of residues. This perhaps implies the importance of this particular region in subunit interactions in this lectin. In addition, many of the residues, involved in carbohydrate binding in legume lectins, appear to be conserved in WBA II. The distinct differences in anomeric specificity observed amongst WBA I, WBA II, ECorL and peanut agglutinin (PNA) may be explained by subtle differences in sequence/structure of their D-loops. WBA II binds adenine quite strongly; a putative adenine binding sequence has been identified. PMID- 10828456 TI - EST comparison indicates 38% of human mRNAs contain possible alternative splice forms. AB - Expressed sequence tag (EST) databases represent a large volume of information on expressed genes including tissue type, expression profile and exon structure. In this study we create an extensive data set of human alternative splicing. We report the analysis of 7867 non-redundant mRNAs, 3011 of which contained alternative splice forms (38% of all mRNAs analysed). From a total of 12572 ESTs 4560 different possible alternative splice forms were detected. Interestingly, 70% of the alternative splice forms correspond to exon deletion events with only 30% exonic insertions. We experimentally verified 19 different splice forms from 16 genes in a total subset of 20 studied; all of the respective genes are of medical relevance. PMID- 10828457 TI - Successful mimicry of a complex viral antigen by multiple peptide insertions in a carrier protein. AB - The antigenic properties of a viral peptide from the surface of foot-and-mouth disease virus particles have been successfully mimicked by multiple insertion in solvent-exposed regions of Escherichia coli beta-galactosidase. By increasing the number of viral peptides per enzyme monomer, the average IC(50) of hybrid proteins in a competitive enzyme-linked immunosorbent assay) have decreased to values close to that presented by natural virions. Moreover, the antigenic diversity of these new recombinant enzymes when measured with different anti virus antibodies has also been largely reduced, indicating a better presentation of the epitopes located in the viral peptide. Although bivalent antibody binding could have been favoured by multiple presentation, conformational modifications of the viral peptide, due to the presence of other insertions or a cooperative antibody binding cannot be excluded. In addition, a multidimensional antigenic analysis have grouped together the multiple-inserted proteins with the native virus, suggesting that increasing the number of insertions could be a good strategy to reproduce the antigenic properties of an immunoreactive peptide in a natural multimeric disposition. PMID- 10828458 TI - Insertion of the full-length calcium channel alpha(1S) subunit into triads of skeletal muscle in vitro. AB - A full-length and a C-terminally truncated form of the calcium channel alpha(1S) subunit can be isolated from skeletal muscle. Here we studied whether full-length alpha(1S) is functionally incorporated into the skeletal muscle excitation contraction coupling apparatus. A fusion protein of alpha(1S) with the green fluorescent protein attached to its C-terminus (alpha(1S)-GFP) or alpha(1S) and GFP separately (alpha(1S)+GFP) were expressed in dysgenic myotubes, which lack endogenous alpha(1S). Full-length alpha(1S)-GFP was targeted into triad junctions and restored calcium currents and excitation-contraction coupling. GFP remained colocalized with alpha(1S), indicating that intact alpha(1S)-GFP was inserted into triads and that the C-terminus remained associated with the excitation contraction coupling apparatus. PMID- 10828459 TI - hKCNMB3 and hKCNMB4, cloning and characterization of two members of the large conductance calcium-activated potassium channel beta subunit family. AB - We cloned two beta subunits of large-conductance calcium-activated potassium (BK) channels, hKCNMB3 (BKbeta1) and hKCNMB4 (BKbeta4). Profiling mRNA expression showed that hKCNMB3 expression is enriched in testis and hKCNMB4 expression is very prominent in brain. We coexpressed BK channel alpha (BKalpha) and BKbeta4 subunits in vitro in CHO cells. We compared BKalpha/beta4 mediated currents with those of smooth muscle BKalpha/beta1 channels. BKbeta4 slowed activation kinetics more significantly, led to a steeper apparent calcium sensitivity, and shifted the voltage range of BK current activation to more negative potentials than BKbeta1. BKalpha/beta4 channels were not blocked by 100 nM charybdotoxin or iberiotoxin, and were activated by 17beta-estradiol. PMID- 10828460 TI - Regulation of [Ca(2+)](i) homeostasis in MRP1 overexpressing cells. AB - Regulation of capacitative Ca(2+) entry was studied in two different multidrug resistance (MDR) protein (MRP1) overexpressing cell lines, HT29(col) and GLC4/ADR. MRP1 overexpression was accompanied by a decreased response to thapsigargin. Moreover, inhibition of capacitative Ca(2+) entry by D, L-threo-1 phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP) was abolished in MRP1 overexpressing cells. Both PDMP and the MRP1 inhibitor MK571 greatly reduced InsP(3)-mediated (45)Ca(2+) release from intracellular stores in HT29 cells. Again, these effects were virtually abolished in HT29(col) cells. Our results point to a modulatory role of MRP1 on intracellular calcium concentration ([Ca(2+)](i)) homeostasis which may contribute to the MDR phenotype. PMID- 10828461 TI - Redox state dependency of HERGS631C channel pharmacology: relation to C-type inactivation. AB - The S631C mutation in human ether-a-go-go-related gene (HERG) channels has previously been reported to disrupt C-type inactivation and ion-selectivity when Cys-631 is in the oxidized state. In this study, we report the relation between pharmacology and C-type inactivation for HERGS631C channels. We demonstrate that HERGS631C in its reduced state is fully blocked by 1 microM astemizole, terfenadine and dofetilide, similar to wild-type HERG channels. In contrast, oxidized HERGS631C is insensitive for these blockers. Our results suggest that an interaction with HERG channels in the inactivated state might be a common mechanism to a variety of drugs known to block HERG channels with high affinity. PMID- 10828462 TI - Chaperone-like activity of synucleins. AB - Synucleins are a family of small proteins that are predominantly expressed in neurons. The functions of the synucleins are not entirely understood, but they have been implicated in the pathogenesis of several neurodegenerative diseases. Our data show that alpha-, beta- or gamma-synuclein suppresses the aggregation of thermally denatured alcohol dehydrogenase and chemically denatured insulin. The A53T but not the A30P mutant alpha-synuclein was able to inhibit the aggregation of insulin and the chaperone-like activity of alpha-synuclein was lost upon removal of its C-terminal residues 98-140. These results demonstrate that synucleins with the exception of the A30P mutant possess chaperone-like activity. PMID- 10828463 TI - Multirecording of Ca(2+) signals from inner retinal neurons evoked by light stimulation of photoreceptors. AB - We simultaneously monitored changes of intracellular free Ca(2+) concentration ([Ca(2+)](i)) following different light stimuli from different inner retinal neurons of the turtle retina slice preparation. [Ca(2+)](i) increased with an increase of the light stimulus intensity. Some of the cells also showed color opponent Ca(2+) signals. 2-Amino-4-phosphonobutyric acid (APB) blocked in particular [Ca(2+)](i) increases and picrotoxin enhanced the observed [Ca(2+)](i) changes. These data support the idea that the observed [Ca(2+)](i) changes result from light stimulation and subsequent retinal processing. Similar Ca(2+) signals were observed when the release of Ca(2+) from internal stores was blocked with caffeine and thapsigargin. These results indicate that retinal Ca(2+) signals evoked by light stimulation depend to a large extent on voltage-dependent Ca(2+) influx and might therefore reflect signal processing. PMID- 10828464 TI - Representation of stereoscopic edges in monkey visual cortex. AB - Form perception in random-dot stereograms is based on information that resides in the correlation between the two images, but is not present in either image alone. We have studied the coding of stereoscopic figures in the neural activity of areas V1 and V2 of alert behaving monkeys. While cells in V1 generally responded according to the disparity of the surface at the receptive field, we found cells in area V2 that responded selectively to the figure edges. These cells signaled the location and orientation of contrast borders as well as stereoscopic edges, and were often selective for the direction of the step in depth. We concluded that stereoscopic edges are explicitly represented in area V2. PMID- 10828465 TI - Surface integration influences depth discrimination. AB - Image fragments arising from partial occlusion may be perceptually unified by a surface integration process on the basis of similar color or texture. In a new objective measure pitting surface feature similarity against binocular disparity, observers discriminated whether a colored circle had either crossed or uncrossed disparity relative to a surrounding gray rectangle. Sensitivity to disparity was impaired only when (1) the configuration of the other surface fragments in the display supported the integration of a surface behind the rectangle and circle, and (2) matched the color of the central circle. Results were consistent with the hypothesis that a surface integration process integrated similarly-colored surface fragments into a smooth surface, even when those fragments were at different depths. Surface integration caused small and reliable effects on depth perception despite unambiguous disparity information. Perceived depth does not depend solely upon disparity, and may be determined after three-dimensional figural unity is established. PMID- 10828466 TI - Direction biasing by brief apparent motion stimuli. AB - The perceived direction of a motion step (probe stimulus) can be influenced by an earlier motion step or a brief motion sweep containing a series of steps (biasing stimulus). Depending upon experimental conditions, the biasing of the direction of the probe step (a phase shift of 180 degrees +/-Phi) by a biasing stimulus which precedes it by approximately 250 ms can either increase (positive filter biasing) or decrease (negative filter biasing) the tendency to see the probe move in the biasing direction as computed with a motion filter with a biphasic temporal impulse response. In a series of experiments it was found that biasing motions traversing 90 degrees of phase angle in fewer than six steps in less than 100 ms produced positive filter biasing. Also, biasing of the probe direction could be dissociated from the consciously reported direction of the biasing stimulus, and it did not occur when the probe preceded rather than followed the biasing stimulus. A biasing sweep containing more than six steps traversing 90 degrees or a sweep traversing 270 degrees produced negative filter biasing. Perceptual fusion of the steps of the sweep was not a necessary condition for obtaining negative filter biasing. In general, the negative filter biasing effects were found to be the most pervasive for the conditions investigated, and they are suggestive of a direction-specific, adaptation-like (gain-control) process in first-order motion filters. The exception to the negative biasing rule was found only with biasing stimuli which were short in duration or distance spanned. PMID- 10828467 TI - Absence of a chromatic linear motion mechanism in human vision. AB - We have investigated motion mechanisms in central and perifoveal vision using two frame random Gabor kinematograms with isoluminant red-green or luminance stimuli. In keeping with previous results, we find that performance dominated by a linear motion mechanism is obtained using high densities of micropatterns and small temporal intervals between frames, while nonlinear performance is found with low densities and longer temporal intervals [Boulton, J. C., & Baker, C. L. (1994) Proceedings of SPIE, computational vision based on neurobiology, 2054, 124-133]. We compare direction discrimination and detection thresholds in the presence of variable luminance and chromatic noise. Our results show that the linear motion response obtained from chromatic stimuli is selectively masked by luminance noise; the effect is selective for motion since luminance noise masks direction discrimination thresholds but not stimulus detection. Furthermore, we find that chromatic noise has the reverse effect to luminance noise: detection thresholds for the linear chromatic stimulus are masked by chromatic noise but direction discrimination is relatively unaffected. We thus reveal a linear 'chromatic' mechanism that is susceptible to luminance noise but relatively unaffected by color noise. The nonlinear chromatic mechanism behaves differently since both detection and direction discrimination are unaffected by luminance noise but masked by chromatic noise. The double dissociation between the effects of chromatic and luminance noise on linear and nonlinear motion mechanisms is not based on stimulus speed or differences in the temporal presentations of the stimuli. We conclude that: (1) 'chromatic' linear motion is solely based on a luminance signal, probably arising from cone-based temporal phase shifts; (2) the nonlinear chromatic motion mechanism is purely chromatic; and (3) we find the same results for both perifoveal and foveal presentations. PMID- 10828468 TI - Velocity discrimination in scotopic vision. AB - To characterize scotopic motion mechanisms, we examined how variation in average luminance affects the ability to discriminate velocity. Stimuli were drifting horizontal sine-wave gratings (0.25, 1.0 and 2.0 c/deg) viewed through a 2 mm artificial pupil and neutral density filters to produce mean adapting levels from 2.5 to -1.5 log photopic trolands. Drift temporal frequency varied from 0.5 to 36.0 Hz. Grating contrasts were either three or five times direction discrimination threshold contrasts at each adaptation level. Following 30 min adaptation, two drifting gratings were presented sequentially at the fovea. Subjects were asked to indicate which interval contained the faster moving stimulus. The Weber fraction for each base temporal frequency was determined using a staircase method. As previously reported, velocity discrimination performance was most acute at temporal frequencies of about 8.0 Hz and greater than 20.0 Hz (though there are individual differences), and fell off at both higher and lower temporal frequencies under photopic conditions. As adaptation level decreased, discrimination of high temporal frequencies in the central retina became increasingly worse, while discrimination of low temporal frequencies remained largely unaltered. The overall scotopic discrimination performance was best at about 3.0 Hz. These results can be explained by a motion mechanism comprising both low-pass and band-pass temporal filters whose peak and temporal cut-off shifts to lower temporal frequencies under scotopic conditions. PMID- 10828469 TI - The dependence of motion repulsion and rivalry on the distance between moving elements. AB - We investigated the extent to which motion repulsion and binocular motion rivalry depend on the distance between moving elements. The stimuli consisted of two sets of spatially intermingled, finite-life random dots that moved across each other. The distance between the dots moving in different directions was manipulated by spatially pairing the dot trajectories with various precisions. Data from experiment 1 indicated that motion repulsion occurred reliably only when the average distance between orthogonally moving elements was at least 21.0 arc min. When the dots were precisely paired, a single global direction intermediate to the two actual directions was perceived. This result suggests that, at a relatively small spatial scale, interaction between different directions favors motion attraction or coherence, while interaction at a somewhat larger scale generates motion repulsion. Similarly, data from experiment 2 indicated that binocular motion rivalry was significantly diminished by spatially pairing the dots, which moved in opposite directions in the two eyes. This supports the recent proposal that rivalry occurs at or after the stage of binocular convergence, since monocular cells could not have directly responded to our interocular pairing manipulation. Together, these findings suggest that the neural mechanisms underlying motion perception are highly sensitive to the fine spatial relationship between moving elements. PMID- 10828470 TI - Spatial structure, contrast polarity and motion integration. AB - It has been shown that in the initial stages of motion processing, the ON and OFF pathways stay more or less separated. There is evidence that this distinction between motion signals from opposite contrast polarities remains at least partly intact in the integration stage of local motion information. At the same time, interactions between the two systems are also apparent. Here we constructed stimuli that contained a constant number of moving checks. The checks were either assigned only one contrast polarity, or contrast polarity was distributed across the checks either randomly or evenly. We investigated how the spatial configuration of the moving stimulus affected direction discrimination thresholds for the different polarity distributions. Our results provide new evidence for contrast-sign-specific integration of local motion signals within areas of limited size, and inhibitory interactions between these separate ON and OFF motion sensor pools. PMID- 10828471 TI - Correlates of figure-ground segregation in fMRI. AB - We investigated which correlates of figure-ground-segregation can be detected by means of functional magnetic resonance imaging (fMRI). Five subjects were scanned with a Siemens Vision 1.5 T system. Motion, colour, and luminance-defined checkerboards were presented with alternating control conditions containing one of the two features of the checkerboard. We find a segregation-specific activation in V1 for all subjects and all stimuli and conclude that neural mechanisms exist as early as in the primary visual cortex that are sensitive to figure-ground segregation. PMID- 10828472 TI - On the linearity of accommodation dynamics. AB - Available experimental data on the gain and phase lags in the closed-loop accommodation responses to stimuli whose dioptric vergence changes sinusoidally with time are examined. It is shown that both phase lags and gain change almost linearly with the temporal frequency of the stimulus. This implies that the phase lags correspond to a frequency-independent time delay and that, if the system is linear, the impulse response should take a symmetrical (sin x/x)(2) form, with a corresponding standard form of step response. Comparison of this derived step response with the quasi-exponential results found experimentally underlines the limitations of attempting to generalize dynamic accommodation performance from responses obtained with predictable sinusoidally-changing stimuli and the application of a linear model. Consideration of the frequency response which would lead to an exponential step response supports the argument that, although the gain results obtained with sinusoidal stimuli are reasonably representative of those applying with other forms of stimulus, the predictable nature of sinusoidally-changing stimuli makes the derived phase lags inapplicable to non predictable step or other stimuli. Other limitations in linear models of the response are discussed. PMID- 10828473 TI - The predictive utility of the Teller acuity cards for assessing visual outcome in children with preterm birth and associated perinatal risks. AB - We assessed binocular grating visual acuity with the Teller acuity cards (TAC) in 3-36-month-olds at risk for visual disorders. After 3-8 years, each child was assessed with the TAC and with a battery of tests of spatial and non-spatial vision. The initial TAC score: (1) was uncorrelated with any of the later measures; (2) had low positive, but high negative predictive value for the later tests; (3) had low sensitivity, but high specificity for identifying children with and without visual disorders, respectively. We concluded that early TAC grating acuity predicts visual outcome, but perhaps only for children with initially normal grating acuity. PMID- 10828474 TI - Visual-vestibular interaction in progressive supranuclear palsy. AB - We measured the stability of gaze during horizontal head rotations at 1-3 Hz in four patients with progressive supranuclear palsy (PSP), while they viewed a stationary target. Median gain of compensatory eye movements was 0.94, similar to control subjects. During rotation in darkness, median gain of vestibulo-ocular reflex (VOR) was 0.88, similar to controls. Conversely, the median gain of smooth pursuit eye movements at 1.0 Hz was 0.23, lower than controls. A simple superposition model of smooth pursuit and the VOR could not account for the observed gaze stability during fixation. Our results are further evidence that a visually mediated mechanism, independent of smooth pursuit, optimizes eye movements to compensate for head rotations. PMID- 10828475 TI - Short-chain fatty acid enemas stimulate plasminogen activator inhibitor-1 after abdominal aortic graft surgery: a double-blinded, placebo-controlled study. AB - Colonic ischaemia may complicate aortic graft surgery with high mortality from associated colonic necrosis. Loss of the mucosal barrier function due to ischaemia may promote translocation of endotoxins with secondary systemic disseminated coagulation leading to multiple organ failure. Short-chain fatty acids (SCFAs) stimulate the microcirculation in the human rectum. The aim of this study was to investigate whether SCFA enemas influence systemic endotoxinaemia and fibrinolytic activity during and after elective aortic graft surgery for arteriosclerosis. Thirty-two patients were randomized to SCFA or placebo enemas twice daily from the day before surgery to 7 days after. Blood samples for endotoxin, plasminogen activator inhibitor-1 (PAI-1) activity, tissue-type plasminogen activator (t-PA) antigen, and cross-linked fibrin degradation products (XL-FDP) were drawn before, during, and 7 days after surgery. Four patients, two in each treatment group, developed postoperative endotoxinaemia. PAI-1 was significantly higher on days 2 and 4 in SCFA-treated patients, whereas t-PA was comparable Petween the groups. During the postoperative course, a progressive and near-identical XL-FDP increase was found in the two groups. In elective aortic graft surgery for arteriosclerosis, SCFA enemas likely stimulate systemic PAI-1 activity by promoting colonic tissue reperfusion following aortic unclamping. Endotoxinaemia and fibrinolytic shutdown are uncommon findings. PMID- 10828476 TI - Platelet serotonergic mechanisms in patients with cancer of the urinary bladder. AB - Platelet reactivity of hypertensive patients and patients with cancer of the urinary bladder was examined in vitro in comparison with normotensive patients. A significant increase in free serotonin in the plasma of patients with cancer of the bladder and a less pronounced increase in hypertensive subjects was observed. The platelets in neoplastic disease and hypertension showed a significant reduction in the kinetics of serotonin uptake. However, the release reaction was markedly higher, compared with healthy volunteers. Enhancement of platelet aggregation with serotonin was much greater in patients with cancer of the bladder than in control groups and in hypertensive patients. These data indicate increased sensitivity to serotonin within the platelets of patients with cancer of the urinary bladder. PMID- 10828477 TI - Tissue factor pathway inhibitor levels in patients with homocystinuria. AB - Thrombotic events are a well-recognized complication of homocystinuria. However, the mechanisms involved in the atherogenic and thrombotic effects of homocyst(e)ine remain incompletely understood. The objective of this study was to determine the role of endothelial cell activation/damage as indicated by levels of thrombomodulin, tissue factor and tissue factor pathway inhibitor, and factor VII activity in patients with homocystinuria. Six patients with homocystinuria, nonresponsive to pyridoxine, treated only with trimethylglycine (betaine) were injected with a bolus of 20 IU/kg body weight of unfractionated commercial heparin to induce the release of tissue factor pathway inhibitor from the vascular endothelium. Tissue factor, thrombomodulin, and factor VII activity were measured by enzyme-linked immunosorbent assay and clotting assay before heparin administration. Tissue factor pathway inhibitor antigen and activity were measured before and 5 minutes after the bolus of heparin. Levels of homocyst(e)ine were elevated (patients: 144.2+/-19.2 micromol/L; controls: 10.2+/ 0.9 micromol/L); however, levels of thrombomodulin, tissue factor, and tissue factor pathway inhibitor antigen were not statistically different from the control group. In contrast, tissue factor pathway inhibitor activity showed a significantly increased level (patients: 2.09+/-0.34 U/L; controls: 1.14+/-0.20 U/L; p<0.05) that was correlated with homocyst(e)ine. Factor VII activity was significantly decreased (patients: 64.7+/-5.1%; controls: 91.4+/-4.7%; p<0.05) and inversely correlated with homocyst(e)ine. After heparin the patients released higher amounts of tissue factor pathway inhibitor antigen and activity compared with the control group; however, the difference was not statistically significant. Although not treated with antithrombotic drugs, none of the patients had any thromboembolic complications after starting betaine. In addition to betaine treatment, the enhanced factor pathway inhibitor antigen activity observed in this small series of patients suggests that factor pathway inhibitor antigen may play an additional, as yet unexplained, role in this genetic disorder. PMID- 10828478 TI - Nutritional status influences plasma fibrinogen concentration: evidence from the THUSA survey. AB - Nutritional status and risk factors for chronic diseases, including plasma fibrinogen and its determinants, of Africans in the Northwest Province of South Africa, have been studied in a cross-sectional survey. A representative sample of 1854 "apparently healthy" African men and women volunteers aged 15 years and older was recruited from 37 randomly selected sites throughout the Province and stratified for level of urbanisation. Information was collected using validated and culture-sensitive questionnaires. Fasting blood samples were drawn, and all measurements were done with standardised methodology using appropriate equipment, procedures, and controls. Fibrinogen concentration was measured in citrated plasma with the method of Clauss, using the ACL200 automated system and the international fibrinogen standard. The results revealed a population with a high mean plasma fibrinogen (3.17+/-1.10 g/L for HIV-negative men and 3. 64+/-1.12 g/L for HIV-negative women). Factors known to influence plasma fibrinogen, such as age, gender, smoking habit, and physical activity, were also observed in this population. Young rural men and women had the lowest fibrinogen level. Nasal snuff taking and HIV infection did not influence fibrinogen concentration. Multivariate analyses revealed that lower plasma fibrinogen was associated with low to normal body mass index in women, and with dietary intakes compatible with prudent dietary guidelines in men and women (low intakes of animal protein; trans fatty acids and higher intakes of plant protein; dietary fibre, vitamin E, and iron, and a high dietary P/S ratio). Subjects in the higher quartiles of plasma fibrinogen had significantly lower iron, vitamin E, and vitamin B6 (women) status. Increases in fibrinogen were associated with significant increases in serum lipids. Both under- and overnutrition seem to be associated with high plasma fibrinogen. It is concluded that overall nutritional status, possibly in addition to specific nutrients (and foods), influences plasma fibrinogen. PMID- 10828479 TI - Inactivation of platelet PDE2 by an affinity label: 8-[(4-bromo-2, 3 dioxobutyl)thio]cAMP. AB - Cyclic GMP-stimulated cyclic nucleotide phosphodiesterase (PDE2) is the second most abundant of this class of enzymes in platelets. PDE2 probably plays an important role in the regulation of elevated intracellular concentrations of cAMP and cGMP in platelets inhibited by prostacyclin and/or nitric oxide. The cAMP and cGMP PDEs have catalytic domains with 28-40% identity, but vary in their substrate specificity and affinity. As a first step toward the goal of identifying important amino acids in the substrate binding site pocket, we have employed the affinity analog 8-[(4-bromo-2, 3-dioxobutyl)thio]adenosine-3'5' cyclic monophosphate (8-BDB-TcAMP) to inactivate PDE2 and observe the pattern of protection by substrates and their products. Incubation of purified platelet PDE2 with 8-BDB-TcAMP (2-10 mM) resulted in a time-dependent, irreversible inactivation of the enzyme with a second-order rate constant of 0.013 min(-1) mM( 1). Both substrates, cAMP and cGMP, as well as the products of hydrolysis by PDE2, AMP and GMP, exhibited concentration-dependent protection against inhibition by 8-BDB-TcAMP, but no protection was noted with ADP or ATP, which are not hydrolyzed by the enzyme. This compound, 8-BDB-TcAMP, and similar affinity reagents should prove useful in delineating amino acids in the active site of PDE2. PMID- 10828480 TI - Human platelet aggregation is not altered by Shiga toxins 1 or 2. AB - The hemolytic uremic syndrome involves the presence of Shiga toxin producing strains of Escherichia coli and is associated with thrombocytopenia, platelet activation, and microthrombi formation. We have, therefore, investigated the ability of Shiga toxin isotypes 1 and 2 to cause or enhance platelet aggregation under resting or arterial-flow conditions using a sensitive quenched-flow system and single-particle counting. Incubation of platelets with Shiga toxins 1 or 2 at 10(-10) M or 10(-9) M for 0.5-2 hours failed to induce platelet aggregation under static or physiological flow conditions, either by themselves or in the presence of ADP or thrombin. Thus, these Shiga toxins do not appear to be able to influence platelet function directly, and their ability to cause platelet thrombi in vivo must result from indirect mechanisms. PMID- 10828481 TI - Chloroquine: a multipotent inhibitor of human platelets in vitro. AB - Chloroquine inhibited human platelet aggregation in vitro both at receptor- and nonreceptor-operated stimuli. The inhibition was dose-dependent, recorded on isolated platelets as well as in platelet-rich plasma, and followed the rank order of stimuli: adrenaline (second phase)>phorbol 12-myristate 13 acetate>adenosine diphosphate>adrenaline (first phase)>thrombin>calcium ionophore A23187. In thrombin-activated platelets, chloroquine decreased in a dose dependent manner phospholipase A(2)-induced arachidonic acid liberation from membrane phospholipids, malondialdehyde formation (a marker of membrane phospholipid peroxidation), and thromboxane generation, considered the most potent autoaggregatory agent. Chloroquine only slightly altered the arachidonic acid cascade of platelets stimulated with A23187 and phorbol 12-myristate 13 acetate. Histamine formation and liberation induced with thrombin and A23187 were not affected by chloroquine. On the other hand, thrombin-stimulated serotonin secretion was significantly decreased with chloroquine in the concentration of 10 micromol/L. This indicated that chloroquine might interfere with stimulated secretion from platelets. The results suggest that chloroquine inhibited activated platelets: first, intracellularly; second, in a close relationship to the intraplatelet Ca(2+) mobile pool; and third, most probably at the site of platelet phospholipase A(2) activation. PMID- 10828482 TI - Different role of platelet glycoprotein GP Ia/IIa in platelet contact and activation induced by type I and type III collagens. AB - The role of glycoprotein Ia/IIa was studied during platelet contact and aggregation induced by type I and type III collagen. The anti-glycoprotein Ia/IIa (6F1) antibody inhibited type I collagen-induced aggregation but did not inhibit the first contact between platelets and collagen. In contrast, it was without effect either on type III collagen-induced contact or platelet interaction with the subendothelium in a static assay. Platelet aggregation induced by type III collagen was only slightly slowed down by 6F1 but pp72 spleen tyrosine kinase phosphorylation was not modified even at concentrations of 6F1 that completely blocked platelet activation induced by type I collagen. Our results indicate that glycoprotein Ia/IIa is not a primary binding site for type I or type III collagen on the platelet membrane. This receptor is more specifically involved in type I collagen-induced platelet spreading and aggregation. PMID- 10828483 TI - PLATSAK, a potent antithrombotic and fibrinolytic protein, inhibits arterial and venous thrombosis in a baboon model. AB - Antiplatelet-antithrombin-staphylokinase (PLATSAK) is a chimeric protein that was recombinantly produced in Escherichia coli cells. The protein was designed to target haemostasis at three different levels. It consists of staphylokinase for activation of fibrinolyis, the Arg-Gly-Asp sequence for the prevention of platelet aggregation, and an antithrombotic peptide for the inhibition of thrombin. The in vivo activity of PLATSAK was evaluated by assessing its effect on platelet deposition in a baboon model of arterial and venous thrombosis. Dacron vascular graft segments and expansion chambers, inserted as extensions into permanent femoral arteriovenous shunts, were used to simulate arterial and venous thrombosis, respectively. PLATSAK (3.68 mg/kg) was administered as a bolus 10 minutes before placement of the thrombogenic devices. Platelet deposition onto the graft surface and in the expansion chamber was imaged in real time with a scintillation camera as the deposition of 111In-labeled platelets. After 2 hours, platelet deposition in the graft segments and expansion chambers was inhibited by 50% and 85%, respectively, when compared to control studies. The activated partial thromboplastin time was lengthened to greater than 120 seconds. Interestingly, the level of fibrinogen degradation products in plasma did not increase after administration of PLATSAK. These results demonstrate that PLATSAK effectively inhibited platelet deposition in both arterial- and venous-type thrombosis in an animal model. PMID- 10828484 TI - Mechanisms involved in the augmentation of arachidonic acid-induced free-radical generation from rat neutrophils following hypoxia-reoxygenation. AB - Polymorphonuclear leukocytes are known to play an important role in hypoxia/ischemia and reoxygenation injury. The present study was undertaken to investigate the involvement of protein kinase C, calmodulin, and cyclic adenosine monophosphate in the augmentation of the free-radical generation observed after hypoxia-reoxygenation (H-R). Free-radical generation from the rat polymorphonuclear leukocytes was measured as the arachidonic acid (1-5x10(-5) M) induced luminol-dependent chemiluminescence response, which was augmented following H-R. The increase in free-radical generation after H-R was completely blocked by the pretreatment of cells with PKC inhibitor H(7), whereas indomethacin (a cyclo-oxygenase inhibitor) or forskolin (an adenylate cyclase activator) failed to modulate the H-R-dependent response. However, W(7)-a calcium/calmodulin (Ca(2+)/CaM) antagonist-partially reduced the augmented free radical generation observed in the H-R cells. Results obtained thus suggest the possible involvement of protein kinase C and calcium in the augmentation of the free-radical generation response following H-R. PMID- 10828485 TI - Transforming growth factor-beta(1) restores antiplatelet function of endothelial cells exposed to anoxia-reoxygenation injury. AB - Transforming growth factor-beta(1) released from platelet alpha-granules may preserve endothelial functions in injured vessels. However, direct evidence is lacking regarding how this cytokine modifies the antithrombotic function of injured endothelial cells. We performed an in vitro study to investigate the effects of transforming growth factor-beta(1) on platelet functions in the presence of cultured endothelial cells exposed to anoxia-reoxygenation injury. Cultured bovine aorta endothelial cells were placed in an anoxic chamber (0.5% O(2), 5% CO(2)) for 60 minutes followed by a 90-minute reoxygenation. Collagen (2 microg/mL)-induced platelet aggregation (10(8) platelets/mL), as determined by impedance aggregometry, was potently inhibited in the presence of control endothelial cells (17.4+/-3.3 Omega) at a concentration of 5x10(4) cells/mL, as compared to their absence (68. 2+/-2.2 Omega). Inhibition of platelet aggregation was attenuated in endothelial cells exposed to anoxia-reoxygenation (54.6+/-2.5 Omega). However, preincubation of endothelial cells with transforming growth factor-beta(1) (1.0 ng/mL) for 16 hours partially recovered the inhibitory capability of platelet aggregation by injured endothelial cells (40.6+/-3.8 Omega). Cell viability, confirmed by a trypan blue dye exclusion test, was similar (93-96%), including control, 1.0 ng/mL transforming growth factor-beta(1) and/or anoxia-reoxygenation-pretreated cells. The capability of platelet inhibition was restored when the endothelial cells were preincubated for 4 hours or more. Restoration of antiplatelet capacity in endothelial cells by transforming growth factor-beta(1) was suggested to be due to several mechanisms, including an increase in nitric oxide synthase activity, up-regulation of prostacyclin release, and restoration of adenosine triphosphate diphosphohydrolase activity, which was attenuated by anoxia-reoxygenation pretreatment. In summary, transforming growth factor-beta(1) released from activated platelets may play a compensatory role in the preservation of endothelial functions to inhibit platelet activation. PMID- 10828486 TI - Platelet-derived microparticles stimulate coronary artery smooth muscle cell mitogenesis by a PDGF-independent mechanism. AB - This study investigates the role of platelet-derived microparticles for vascular smooth muscle cell (SMC) proliferation. Microparticles concentration dependently stimulated p42/p44 MAP kinase phosphorylation, c-fos induction, DNA synthesis, and proliferation of cultured bovine coronary artery SMC. The maximum mitogenic effects of microparticles were significantly higher than those of platelet derived growth factor (PDGF)-BB. Microparticle-induced SMC mitogenesis was heat sensitive, whereas the effects of PDGF were not. In addition, neutralizing anti PDGF antibodies prevented PDGF-induced DNA synthesis but did not inhibit the effects of microparticles. In contrast to PDGF, which potently stimulated SMC migration, microparticles had only minor migratory activity. These results demonstrate a novel mechanism of SMC mitogenesis by platelet-derived microparticles that is probably independent of PDGF. PMID- 10828487 TI - Advances in understanding neuronal somatostatin receptors. AB - It has long been considered that somatostatin acts as a neuromodulator in the mammalian central nervous system but its precise physiological roles remain elusive. Early studies to identify somatostatin-binding sites revealed a widespread heterogeneous pattern, especially in the CNS. More recently, a family of somatostatin receptors have been identified, of which five genes (sst(1-5)) have been cloned. In this review, we discuss current data describing the localisation of the five receptor types. Recent progress in understanding their function has been made using high-affinity, selective receptor ligands and transgenic animal technology. Finally, the therapeutic potential for somatostatin receptor-selective compounds as analgesics is considered. PMID- 10828488 TI - Reactive oxygen species and vascular cell apoptosis in response to angiotensin II and pro-atherosclerotic factors. AB - Reactive oxygen species (ROS) are known to induce apoptotic cell death in various cell types. In the vessel wall, ROS can be formed by macrophages within the atherosclerotic plaque or can act on the endothelium after adhesion of monocytes or leucocytes. Moreover, ROS are endogenously synthesized by endothelial and vascular smooth muscle cells by NAD(P)H oxidase. Enhanced ROS production is a very early hallmark in the atherogenic process, suggesting a link between ROS and apoptosis. In endothelial cells, the endogenous generation of ROS is induced by different pro-inflammatory and pro-atherosclerotic factors such as Ang II, oxLDL or TNFalpha, which all promote the execution of programmed cell death. ROS synthesis is thereby causally involved in apoptosis induction, because antioxidants prevent endothelial cell death. The pro-apoptotic effects of endogenous ROS in endothelial cells mechanistically seems to involve the disturbance of mitochondrial membrane permeability followed by cytochrome c release, which finally activates the executioner caspases. In contrast to the pro apoptotic capacity of ROS in endothelial cells, in vascular smooth muscle cells emerging evidence suggests that endogenous ROS synthesis promotes cell proliferation and hypertrophy and does not affect cell survival. However, high concentrations of exogenous ROS can also stimulate smooth muscle cell apoptosis as shown for other cell types probably via activation of p53. Taken together, the double-edged effects of endogenously derived ROS in endothelial cells versus VSMC may provide a mechanistic clue to the anti-atherosclerotic effects of antioxidants shown in experimental studies, given that the promotion of endothelial survival in combination with inhibition of VSMC proliferation blocks two very important steps in the pathogenesis of atherosclerosis. PMID- 10828489 TI - New frontiers in the biology of GLP-2. AB - Glucagon-like peptide-2 (GLP-2) is a 33 amino acid peptide hormone released from the intestinal endocrine cells following nutrient ingestion. GLP-2 exerts trophic effects on the small and large bowel epithelium via stimulation of cell proliferation and inhibition of apoptosis. GLP-2 also upregulates intestinal glucose transporter activity, and reduces gastric emptying and gastric acid secretion. The activity of GLP-2 is regulated in part via renal clearance and cleavage by the aminopeptidase dipeptidyl peptidase IV. In experimental models of intestinal disease, GLP-2 reversed parenteral nutrition-induced mucosal atrophy and accelerated the process of endogenous intestinal adaptation in rats following major small bowel resection. GLP-2 also markedly attenuated intestinal injury and weight loss in mice with chemically-induced colitis, and significantly reduced mortality, bacterial infection and intestinal mucosal damage in mice with indomethacin-induced enteritis. The actions of GLP-2 are transduced by a recently cloned glucagon-like peptide-2 receptor (GLP-2R) that represents a new member of the G protein-coupled receptor superfamily. The GLP-2R is expressed in a highly tissue-specific manner predominantly in the gastrointestinal tract and GLP-2R activation is coupled to increased adenylate cyclase activity. The available evidence suggests that the biological properties of GLP-2 merit careful therapeutic assessment in selected human diseases characterized by injury and defective repair of the gastrointestinal epithelium. PMID- 10828490 TI - Free-ligand accelerated dissociation of insulin-like growth factor 1 (IGF-1) from the type I IGF receptor is reduced by insulin-like growth factor binding protein 3. AB - Insulin-like growth factor binding proteins (IGFBP) can inhibit or accentuate the mitogenic activities of insulin-like growth factor 1 (IGF-1) depending upon the experimental model employed. Inhibitory effects may be attributed to sequestration of IGF-1 onto IGFBP rather than the type I IGF receptor. We have demonstrated that the presence of IGFBP in a simple equilibrium binding assay significantly reduces the total amount of IGF-1 bound to the type I IGF receptor and increases the IC(50) for IGF-1 binding. On the basis of such an experiment, performed at equilibrium, IGFBP should reduce the mitogenic activity of IGF-1. Recent work has demonstrated an inverse correlation between the dissociation rate of insulin-like molecules from their receptors and their mitogenic activity. It has also been suggested that the increased rate of dissociation of insulin and IGF-1 from their receptors at increased ligand concentrations serves as a 'dampening' mechanism to decrease mitogenic signalling. We have demonstrated increased rates of dissociation of IGF-1 from the type I IGF receptor with increasing concentrations of IGF-1. Furthermore, IGFBP-3 inhibits the acceleration of dissociation rates due to increased IGF-1 levels. Thus, under receptor saturating conditions IGFBP-3 may act to increase mitogenesis by increasing the residence time of individual molecules of IGF-1 upon the type I IGF receptor. PMID- 10828491 TI - Calcitonin gene-related peptide regulates amino acid absorption across rat jejunum. AB - The calcitonin gene related peptide (CGRP) is widely distributed in the enteric nervous system and gut afferents. Its role in normal digestion and absorption is not characterised. This study is conducted to elucidate whether CGRP regulates amino acid absorption in the small intestine. In in vivo experiments using the single-pass perfusion technique, intravenous infusion of CGRP (250-750 pmol/kg min) reduced alanine absorption by 35-40%. The effects were completely blocked by the antagonist hCGRP (8-37). Moreover, intravenous infusion of CGRP antagonist blocked the inhibitory effect of intraluminal capsaicin perfusion on alanine absorption. Similarly, intracerebral injection of CGRP decreased alanine absorption, an effect which was reduced by vagotomy. In vitro experiments using isolated jejunal strips showed that CGRP reduced alanine absorption in a dose dependent manner. At 6 pM, CGRP decreased alanine absorption by 33%. Similarly, CGRP reduced the absorption of proline and taurine by 20 and 11.5%, respectively. Kinetic studies revealed that CGRP reduces alanine influx into intestinal epithelial cells by inhibiting the affinity of the carriers. It is demonstrated that CGRP is involved in the regulation of jejunal amino acid absorption through intrinsic (enteric) and extrinsic (central) neural mechanisms. PMID- 10828492 TI - TGFbeta1 autocrine growth control in isolated pancreatic fibroblastoid cells/stellate cells in vitro. AB - TGFbeta1 is a multifunctional factor, controlling cellular growth and extracellular matrix production. Deletion of the TGFbeta1 gene in mice results in multiple inflammatory reactions. Targeted overexpression of TGFbeta1 in pancreatic islet cells leads to fibrosis of the exocrine pancreas in transgenic mice. In pancreatic fibrosis interstitial fibroblasts are primary candidates for production and deposition of extracellular matrix. Still, little is known about regulation of these cells during development of pancreatic disease. We established primary cell lines of pancreatic fibroblastoid/stellate cells (PFC) from rat pancreas. Investigation of rPFCs in vitro shows TGFbeta1 expression by RT-PCR analysis. Mature TGFbeta1 was detected in culture supernatants by immunoassay. Rat PFCs in culture possess both receptors TGFbeta receptor type I, and type II, necessary for TGFbeta1 signal transduction. Inhibition of TGFbeta1 activity by means of neutralizing antibodies interferes with an autocrine loop and results in a 2-fold stimulation of cell growth. So far, pancreatic fibroblastoid/stellate cells in vitro were known as a target of TGFbeta1 action, but not as a source of TGFbeta1. Our data indicate TGFbeta1 activity in rat pancreas extends beyond regulation of matrix production, but appears to be important in growth control of pancreatic fibroblastoid cells. PMID- 10828493 TI - Purification and characterization of antimicrobial and vasorelaxant peptides from skin extracts and skin secretions of the North American pig frog Rana grylio. AB - Eight peptides with differential growth-inhibitory activity against the gram positive bacterium Staphylococcus aureus, the gram-negative bacterium Escherichia coli and the yeast, Candida albicans were isolated from an extract of the skin of the North American pig frog Rana grylio. The primary structures of these antimicrobial peptides were different from previously characterized antimicrobial peptides from Ranid frogs but on the basis of sequence similarities, the peptides may be classified as belonged to four previously characterized peptide families: the ranatuerin-1, ranatuerin-2 and ranalexin families, first identified in the North American bullfrog, Rana catesbeiana, and the temporin family first identified in the European common frog Rana temporaria. Peptides belonging to the brevinin-1, brevinin-2, esculentin-1, and esculentin-2 families, previously isolated from the skins of other species of Ranid frogs, were not identified in the extracts. The ranatuerin-1 and ranalexin peptides showed broadest spectrum of antimicrobial activity whereas the temporins were active only against S. aureus. Synthetic replicates of temporin-1Gb (SILPTIVSFLSKFL.NH(2)) and temporin-1Gd (FILPLIASFLSKFL.NH(2)) produced concentration-dependent relaxation of preconstricted vascular rings from the rat thoracic aorta (EC(50) = 2.4+/-0.1 microM for temporin-1Gb and 2.3+/-0.2 microM for temporin-1Gd). The antimicrobial peptides that were isolated in extracts of the skin R. grylio were present in the same molecular forms in electrically-stimulated skin secretions of the animal demonstrating that the peptides are stored in the granular glands of the skin in their fully processed forms. PMID- 10828494 TI - Isolation and characterization of putative trefoil peptide receptors. AB - Mammalian trefoil factors (TFFs) constitute a group of three peptides (TFF1, TFF2 and TFF3) widely distributed in the gastrointestinal tract. Although a mucosal protection/healing effect of these peptides is well documented the mechanism of action is still unknown. A mucosal membrane extract was prepared from porcine stomach scrapings and incubated with a gel containing immobilized porcine TFF2. The affinity gel material was specifically eluted with a neutral buffer containing a high concentration of the ligand (porcine TFF2). A subsequent SDS gel electrophoresis showed one protein with a MW of approximately 220 kDa and three proteins with MW around 140 kDa. The proteins were analyzed by trypsin digestion followed by mass spectrometric sequencing of tryptic fragments. In this way a 140-kDa beta subunit of fibronectin receptor and a 224-kDa CRP-Ductin gene product were identified. The CRP-Ductin gene product (also named MUCLIN), which is expressed in the intestinal crypts, is characterized by being a membrane protein with a short cytoplasmic region, a transmembrane domain and a large extracellular region. This protein thus fulfils some of the criteria for being a TFF receptor or a TFF binding protein. PMID- 10828495 TI - Renal content and output of epidermal growth factor in long-term adrenergic agonist-treated rats. AB - This study investigates the renal and urinary levels of epidermal growth factor (EGF) in rats under long-term treatment with alpha- or beta-adrenergic agonists. Urine samples were obtained on days 7, 14 and 21, and renal tissue samples on day 21. EGF was quantified by ELISA and tissue sections were used for immunohistochemistry and in situ hybridization. Fractional kidney weight was increased in the alpha-adrenergic agonist-treated group by 35% when compared with controls. Histological examination of the kidney revealed well-defined wedge shaped areas of tubular dilatations and luminal amorphous material in the distal tubules. Concomitantly, reduced levels of EGF and EGF mRNA were observed, and also the urinary levels of EGF were reduced. Together, these observations indicate alpha-adrenergic treatment to affect the distal tubules. Treatment with the beta-adrenergic agonist did not change fractional kidney weight, but initially the urinary excretion of EGF was reduced. The data add further evidence to the suggestion that activity of the sympathetic nervous system influences renal homeostasis of EGF, either directly or indirectly through renal histopathological changes. PMID- 10828496 TI - The presence of receptors for bombesin/GRP and mRNA for three receptor subtypes in human ovarian epithelial cancers. AB - Bombesin-like peptides can function as autocrine or paracrine growth factors and stimulate the growth of various cancers. The antagonists of bombesin/gastrin releasing peptide (GRP) suppress the proliferation of diverse tumors including ovarian cancer by mechanisms likely mediated by bombesin receptors. In this study, we used the reverse transcription-polymerase chain reaction (RT-PCR) method to evaluate the mRNA expression of three bombesin receptor subtypes: gastrin-releasing peptide receptor (GRPR), neuromedin B receptor (NMBR), and bombesin receptor subtype 3 (BRS-3), in 22 specimens of human epithelial ovarian cancer and in two human ovarian cancer lines. Of the 22 ovarian cancer specimens analyzed, 17 tumors ( approximately 77%) expressed mRNA for GRPR, 19 ( approximately 86%) showed NMBR mRNA and six ( approximately 27%) revealed BRS-3 mRNA. Thus, 14 of 22 specimens ( approximately 64%) expressed mRNAs for both GRPR and NMBR, and five ( approximately 23%) expressed all three subtypes. The expression of GRPR appeared to be greater in poorly differentiated ovarian carcinomas. A higher incidence of BRS-3 expression was observed in samples with tumor Stage IV (4/4, 100%) compared with Stage III (1/17, approximately 6%). mRNA for both GRPR and NMBR was also detected in OV-1063 and UCI-107 human ovarian cancer xenografts, but BRS-3 was found only in OV-1063, which originated from a metastatic tumor. In addition, functional receptors for bombesin/GRP were found in eight of 11 ovarian cancer specimens investigated and in both ovarian cancer lines by receptor binding assay. Our study indicates that GRPR and NMBR are widely distributed in human ovarian carcinomas with BRS-3 being found in Stage IV tumors. Some approaches based on bombesin/GRP receptor antagonists or targeted bombesin analogs could be considered for treatment of ovarian cancers. PMID- 10828497 TI - The blocking effect of BmP02, one novel short-chain scorpion peptide on transient outward K(+) channel of adult rat ventricular myocyte. AB - Two components F-2-7-4 and F-2-7-5, each composed of 28 amino acid residues, were purified from the venom of Buthus martensi Karsch by an opportune procedure with cation-exchange column chromatography and repeated HPLC. Both components were totally accounted to about 0. 88% dry weight of the crude venom. The molecular weights of both components were determined to be 2950 and 2935 by mass spectrometry, which were fully coincidence with that of the known novel short chain peptides BmP02 and BmP03, respectively [Romi-Lebrun R, Martin-Eauclaire M F, Escoubas P, Wu FQ, Lebrun B, Hisada M, Nakajima T. Characterization of four toxins from Buthus martensi scorpion venom, which act on apamin-sensitive Ca(2+) activated K(+) channels. Eur J Biochem 1997;145:457-464]. In addition, the sequence of component F-2-7-4 was analyzed to be the same as that of BmP02. The components F-2-7-4 and F-2-7-5 purified in this study were, thus, finally distinguished to be BmP02 and BmP03 from the same venom. Using whole cell patch clamp recording, it was found that BmP02 diminished the current of transient outward K(+) channel in adult rat ventricular myocyte in a concentration dependent manner. The inhibitory effect was reversible. Dynamic studies showed that the activation, inactivation and recovery processes of the transient outward K(+) channel were not changed significantly after applying of BmP02. In addition, when BmP02 was applied to guinea pig ventricular myocyte, both delayed and inward rectified K(+) currents showed no change compared with the control. The results suggest strongly that BmP02 or -like peptides from scorpion venom may provide a useful probe for the studying of transient outward K(+) channel in rat ventricular myocyte. PMID- 10828498 TI - Central natriuretic peptides regulation of peripheral atrial natriuretic factor release. AB - Atrial natriuretic factor (ANF) and C-type natriuretic peptide (CNP) receptors have been described in encephalic areas and nuclei related to the regulation of cardiovascular as well as sodium and water homeostasis. Stimulation of the anterior ventral third ventricular region of the brain modifies plasma ANF concentration, suggesting the participation of the central nervous system in the regulation of circulating ANF. The aim of this work was to study the effect of centrally applied ANF or CNP on plasma ANF. Normal and blood volume expanded rats (0.8 ml isotonic saline/100 g body weight) were intra cerebralventricularly injected with 1, 10 or 100 ng/microl/min ANF. Blood volume expanded animals were also centrally injected with the same doses of CNP. Blood samples were collected at 5 and 15 min. after intracerebralventricular administration of either ANF or CNP. Centrally applied ANF did not affect circulating ANF in normal blood volume rats. In blood volume expanded animals both ANF (1, 10 or 100 ng/microl/min) and CNP (1 ng/microl/min) decreased plasma ANF concentration after 15 min. Moreover, CNP (10 and 100 ng/microl/min) lowered circulating ANF levels not only at 15 min but also at 5 min. Neither ANF nor CNP elicited any change in mean arterial pressure and heart rate in normal and blood volume expanded rats. These results suggest the existence of a central regulation exerted by natriuretic peptides on circulating ANF levels. Furthermore, this is the first study reporting an effect on plasma ANF induced by centrally applied CNP. PMID- 10828499 TI - Substantial equivalence of antinutrients and inherent plant toxins in genetically modified novel foods. AB - For a safety evaluation of foodstuff derived from genetically modified crops, the concept of the substantial equivalence of modified organisms with their parental lines is used following an environmental safety evaluation. To assess the potential pleiotropic effect of genetic modifications on constituents of modified crops data from US and EC documents were investigated with regard to inherent plant toxins and antinutrients. Analysed were documents of rape (glucosinolates, phytate), maize (phytate), tomato (tomatine, solanine, chaconine, lectins, oxalate), potato (solanine, chaconine, protease-inhibitors, phenols) and soybean (protease-inhibitors, lectins, isoflavones, phytate). In several documents used for notifications no declarations even on essential inherent plant toxins and antinutrients could be found, for instance data on phytate in modified maize were provided only in one of four documents. Significant variations in the contents of these compounds in parental and modified plants especially due to environmental influences were observed: drought stress, for example, was made responsible for significantly increased glucosinolate levels of up to 72.6micromol/g meal in modified and parental rape plants in field trials compared to recommended standard concentrations of less than 30micromol/g. Taking into account these wide natural variations generally the concentrations of inherent plant toxins and antinutrients in modified products were in the range of the concentrations in parental organisms. The results presented indicate that the concept of the substantial equivalence is useful for the risk assessment of genetically modified organisms (GMOs) used for novel foods but possible environmental influences on constituents of modified crops need more attention. Consistent guidelines, specifying data of relevant compounds which have to be provided for notification documents of specific organisms have to be established. Because of the importance of inherent plant toxins and antinutrients on nutritional safety, also coherent databases of standard parental lines and clear criteria for mandatory declarations are necessary. PMID- 10828500 TI - The safety evaluation of phytosterol esters. Part 6. The comparative absorption and tissue distribution of phytosterols in the rat. AB - As part of an extensive safety evaluation programme, a series of studies has been conducted to determine the fate of phytosterols in the rat. Rats were dosed by oral gavage with 14C-labelled samples of cholesterol, beta-sitosterol or beta sitostanol or (3)H-labelled samples of beta-sitostanol, campesterol, campestanol or stigmasterol dissolved in sunflower seed oil. Urine and faeces were collected for up to 96 hours after dosing. There was no quantification of biliary excreted material in these studies. Animals were sacrificed and either prepared for whole body autoradiography or tissues and carcass remains were assayed for 14C or (3)H. The overall absorption of phytosterols was low as judged by tissue and carcass levels of radioactivity. Elimination from the body was mainly in the faeces and was initially very rapid, but traces of material were still being excreted at 4 days after dosing. While total absorption of the phytosterols could not be fully quantified without biliary excretion data, it was clear that cholesterol was absorbed to the greatest extent (27% of the dose in females at 24 hours). Campesterol (13%) was absorbed more than beta-sitosterol and stigmasterol (both 4%) which were absorbed more than beta-sitostanol and campestanol (1-2%). The absorption of phytosterols was slightly greater in females than males. For each test material, the overall pattern of tissue distribution of radioactivity was similar, with the adrenal glands, ovaries and intestinal epithelia showing the highest levels and the longest retention of radioactivity. PMID- 10828501 TI - The development of methods for assessing the in vivo oestrogen-like effects of xenobiotics in CD-1 mice. AB - The increasing awareness and concern about the potential health risks posed to the ecosystem and to man by endocrine disrupting chemicals with oestrogen-like activity in the environment has focused attention on the need for developing sensitive and specific methods for identifying these xenobiotics and to evaluate their degrees of toxic effects. We have conducted dose response studies in immature (21 days old) CD-1 female mice treated with four compounds, diethylstilboestrol (DES) (0.1 microg to 25 mg/kg body weight), alpha-zearalanol (0.5 mg to 25 mg/kg body weight), methoxychlor (0.5 mg to 500 mg/kg body weight) and bisphenol A (10 microg to 100 mg/kg body weight) administered subcutaneously daily for 3 days, and measured a number of uterine markers in treated and control (vehicle treated) mice. These were, in addition to the commonly measured changes in relative uterus weight and histopathological examination of uterine tissue, three other markers indicative of uterotrophic effects, namely, uterine luminal epithelium BrdU labelling index over the last 24 hr, peroxidase activity and lactoferrin expression. All of these markers showed clear dose-related increases in DES- and methoxychlor-treated animals. In the case of alpha-zearalanol treatment, relative uterine weight, peroxidase activity and lactoferrin expression showed dose-related increases at all the doses investigated. BrdU incorporation (an index of cell proliferation) also progressively increased at dose levels ranging from 0.1 mg to 5.0 mg/kg body weight, but apparently decreased at 25 mg/kg body weight. In contrast to these findings, bisphenol-A treatment showed no consistent changes in any of the four markers at the dose levels investigated. Additionally, studies were also conducted on a number of chemicals in CD-1 mice at one dose level. The chemicals investigated were: bisphenol A (1 g/kg body weight/day), naringenin (1 g/kg body weight/day) o,p' DDT (500 mg/kg body weight/day), genistein (1 g/kg/day), coumestrol (0.5 mg/kg/day) and chlordecone (20 mg/kg/day) administered subcutaneously daily for 3 days. There was some variability in response of the markers perhaps indicating that the chemicals did not all act in the same way. The findings of our exploratory in vivo studies in CD-1 mice suggest that the measurement of a range of uterine markers, in addition to organ weight and histopathology, would provide useful information on the potential oestrogenicity of chemicals. PMID- 10828502 TI - 13-Week oral repeated administration toxicity study of bovine lactoferrin in rats. AB - Bovine lactoferrin (LF), which is an iron-binding glycoprotein in milk, was administered orally to groups of 12 males and 12 female rats at dose levels of 200, 600 and 2000mg/kg/day once daily for 13 weeks and its toxicity on repeated administration was examined. Throughout the administration period, there were no deaths caused by administration of the test compound, nor were there any adverse effects noted in the general condition of the animals. The study findings concerning body weight and food consumption, ophthalmology, urinalysis including water consumption, haematology, blood chemistry, necropsy, organ weights and histopathology revealed that there were no apparent changes due to administration of LF. Therefore, the level of LF at which no adverse effect was observed was considered to be 2000mg/kg/day for both sexes. PMID- 10828503 TI - Bioactivation of the food mutagen 2-amino-3-methyl-imidazo[4, 5-f]quinoline (IQ) by prostaglandin-H synthase and by monooxygenases: DNA adduct analysis. AB - 2-Amino-3-methylimidazo[4,5-f]quinoline (IQ) is a known multisite carcinogen in rodents and a potent mutagen in acetyltransferase-proficient Salmonella typhimurium strains on activation by either monooxygenases (MFO) or by prostaglandin H synthase (PHS). The primary metabolites formed by MFO- or PHS mediated IQ-oxidation are different ([Wolz]), but secondary metabolism could ultimately result in the same DNA-binding intermediates. For further investigations, the DNA adduct pattern was now studied by means of (32)P postlabelling analysis in vitro on PHS-activation and compared to that formed on MFO-mediated activation of IQ in hepatocytes. The C8-dG-IQ-adduct N (deoxyguanosin-8-yl)-IQ was the major adduct in all samples, that is, in DNA isolated from S. typhimurium YG1024 treated with PHS-oxidized IQ or its nitro derivative, from ovine seminal vesicle cells, and from hepatocytes exposed to IQ or nitro-IQ. This speaks for the formation of a common DNA-reactive species, presumably an arylnitrenium ion, generated by different pathways in these cellular model systems. The similarity of critical biochemical DNA lesions suggests that PHS can contribute to the bioactivation of IQ in vivo: this is of particular interest in extrahepatic tissues since expression of cytochrome P450 isoenzymes known to be involved in the N-oxidation of IQ is largely confined to the liver. PMID- 10828504 TI - Subchronic (90-day) toxicity study in rats fed gum kondagogu (Cochlospermumgossypium). AB - Although gum kondagogu (Cochlospermum gossypium) is grouped under gum karaya (Sterculia sp.), it differs significantly in terms of physicochemical properties and chemical composition and does not conform to the confirmatory tests prescribed for gum karaya ([Janaki]). Gum karaya has wide applications in the pharmaceutical and food industries, whereas the use of gum kondagogu is yet to be explored. In this context, a short-term toxicity study on gum kondagogu was undertaken in rats. The gum was fed to rats at 0, 0.2%, 1% and 5% (w/w) in feed, for 90 days. Biochemical parameters were measured to assess the toxicity at the end of the study period. The results indicated no significant changes in growth pattern, haematological indices (RBC, WBC, Hb, PCV, MCV, MCH, MCHC, differential leucocyte counts), biochemical analytes (glucose, urea nitrogen, total protein, albumin, bilirubin, creatinine, sodium and potassium ions), activities of plasma and liver enzymes (alkaline phosphatase, alanine amino-transaminase, aspartate aminotransaminase, lactate dehydrogenase, glutathione S-transferase and gamma glutamyl transpeptidases and organ to body mass ratio (brain, heart, lungs, liver, kidneys and spleen). Histopathology of the liver and kidney also did not reveal any abnormality. An increased faecal bulk was observed in rats fed with 5% gum kondagogu. However, faecal moisture content of female rats only was significantly different (P=<0.05) as compared to controls. Thus, it can be inferred, based on the present investigations, that gum kondagogu has a potential application as a food additive, similar to gum karaya. Feeding it at a much higher level (5%) than expected for consumption as a food additive also did not result in any toxic effect. Being non-toxic, gum kondagogu has a potential as a food additive with excellent physicochemical properties and a unique chemical composition. PMID- 10828505 TI - Chemoprevention of 2-acetylaminofluorene-induced hepatotoxicity and lipid peroxidation in rats by kolaviron--a Garcinia kola seed extract. AB - The effect of kolaviron, a mixture of Garcinia biflavonoid 1 (GB1), Garcinia biflavonoid 2 (GB2) and kolaflavanone, used in the treatment of various ailments in southern Nigeria on hepatotoxicity and lipid peroxidation induced by 2 acetylaminofluorene (2-AAF) in rats was investigated. The ability of butylated hydroxyanisole (BHA) to attenuate the toxic effect of 2-AAF was also examined. Kolaviron administered orally to rats at a dose of 100mg/kg body weight twice a day for 1 week before challenge with 2-AAF (200mg/kg feed) and continuously for 3 weeks at a single dose of 200mg/kg body weight reversed the 2-AAF-mediated decrease in final body weight and relative organ weights, especially the liver. BHA was administered at a dose of 7.5g/kg feed to the animals for 4 weeks. The extract decreased significantly the 2-AAF-mediated increase in the activity of aspartate aminotransferase, alanine aminotransferase, gamma-glutamyl transferase and ornithine carbamyl transferase by 58%, 62%, 60% and 67%, respectively. BHA elicited respectively 55%, 63%, 57% and 65% reduction in the 2-AAF induced increase in the activities of these enzymes. Histological examination of the liver slices correlated with the changes in serum enzyme alterations. Similarly, kolaviron decreased the 2-AAF reduction of 5'-nucleotidase and glucose-6 phosphatase activities by 63% and 60%, respectively while BHA elicited 59% and 61% decrease in the activities of these enzymes. Simultaneous administration of kolaviron with 2-AAF inhibited microsomal lipid peroxidation as assessed by the thiobarbituric acid reacting substances (TBARS) formation by 66%. BHA produced a 64% reduction in TBARS formation. In the present study, kolaviron appears to act as an in vivo natural antioxidant and an effective hepatoprotective agent and is as effective as BHA. PMID- 10828506 TI - Differential effects of chronic cyanide intoxication on heart, lung and pancreatic tissues. AB - The histotoxic effects of chronic cyanide insult on heart, lung and pancreatic tissues, and some corroborative enzyme and metabolite changes were studied in New Zealand White rabbits using colorimetric, enzymatic and histochemical methods. Two groups of rabbits were fed for 10 months on either pure growers mash or grower mash +702 ppm inorganic cyanide. There were no significant differences in time-course profiles of serum amylase and fasting blood glucose between the cyanide-fed group and control. Pancreatic islet and heart histologies showed no pathological changes, and there were no significant differences in both serum and heart aspartate transaminase activities between the two groups. However, there were significant decreases (P<0.01) in alkaline phosphatase activity in the lungs of the cyanide-fed group, with corresponding significant (P<0.05) increases in the serum activity of the enzyme. Histological examination of lung tissue of the cyanide-treated rabbits revealed focal areas of pulmonary oedema and necrosis. These results suggest the existence of variabilities in tissue susceptibilities to the toxic effect of chronic cyanide exposure. It would appear that chronic cyanide exposure may not predispose to diabetes in the presence of adequate protein intake. PMID- 10828507 TI - Evaluation of antigen-coating procedures of enzyme-linked immunosorbent assay method for detection of trypanosomal antibodies. AB - Research was undertaken to improve the antigen-coating step of indirect enzyme linked immunosorbent assay (ELISA) method through the use of polystyrene 96-well plates precoated with antigenically stabile crude trypanosomal antigens. The plates were precoated with antigens, air dried and sealed before being packed in plastic bags with silica gel desiccant packets. Such plates stored at +4 and +37 degrees C provided an assay performance, which was superior to that of plates freshly coated with antigens from a frozen stock. Antigen-precoated plates consistently proved stable after storage up to +50 degrees C for at least 1 year. The accuracy of the assay was not affected, i.e. trypanosomal antibody-positive sera were clearly discriminated from trypanosomal antibody-negative negative sera. In contrast, lyophilized trypanosomal antigens lacked stability on storage at +37 degrees C for longer than 1 month. It was concluded that the routine use of antigen precoated polystyrene plates for the enzyme immunoassay technique will contribute to improved assay robustness at an acceptable diagnostic proficiency. The modified coating procedure will also provide an improved quality assurance and standardization procedure for the assay, which is required to allow the reliable detection of trypanosomal antibodies and comparison of data from different laboratories. PMID- 10828508 TI - Neospora caninum seroprevalence and associated risk factors in beef cattle in the northwestern United States. AB - A Neospora caninum seroprevalence and risk factor survey of 2585 cows was conducted in 55 beef cow-calf herds located in five northwestern states of the USA. Blood samples were collected by private veterinary practitioners and management practices were surveyed using a mail questionnaire. Producers were randomly selected from those that employed these veterinarians to perform annual herd pregnancy examinations. Questions were asked about animal management, grazing and feeding, immunization and record keeping practices. Blood was collected from a systematically selected sample of cows in each herd, and age, origin, and pregnancy status were recorded. Blood samples were analyzed for antibodies against N. caninum antigen using a monoclonal antibody-based competitive inhibition (CI) ELISA. Overall seroprevalence was 24% and within herd seroprevalence ranged from 3 to 67% with a median of 19%. Within herd seroprevalence and mean inhibition percentage were different between the five states. Herds that managed their cows on range for summer grazing had lower seroprevalence than those that did not, while increased seroprevalence was associated with higher winter stocking density. Cows less than 3 years of age had higher CI ELISA inhibition percent values than cows greater than 6 years of age. No relationship was noted between serologic status and individual cow origin (purchased or raised), or pregnancy status at the time of sampling. PMID- 10828509 TI - Determination of duration of immunity of calves vaccinated with the Theileria annulata schizont cell culture vaccine. AB - Bovine tropical theileriosis caused by Theileria annulata is a serious haemoprotozoan disease of cattle affecting exotic cattle, their crossbreeds and young indigenous calves. Cell culture vaccines have been developed and used effectively in various countries for the control of this disease. However, the duration of immunity provided by these vaccines is poorly understood. The present experiments were planned to study the duration of immunity in animals after vaccination with the T. annulata (Hisar) schizont cell culture vaccine. Two groups of calves were vaccinated and challenged after a period of 3 and 6 months, respectively. There was no fever in any of the vaccinated calves after challenge. However, the vaccinated animals exhibited mild to moderate enlargement of lymph nodes and parasitological reactions. The parasitological reactions were very mild in calves challenged after 3 months and moderate in calves challenged after 6 months. There was a mild but significant decrease in the haematological values of calves after challenge. A significant rise in the anti-theilerial antibody titres was observed in all calves after vaccination, which increased further, by many folds after challenge. On the other hand, all the challenge control calves showed symptoms of acute theileriosis and died. The observations suggested that the T. annulata (Hisar) schizont cell culture vaccine provided immunity in vaccinated animals for at least 6 months in the absence of field tick challenge. However, there was some decline in immunity after 6 months, if the animals are not exposed to ticks during this period. PMID- 10828510 TI - The Ibizian hound presents a predominantly cellular immune response against natural Leishmania infection. AB - Veterinarians working in the Balearic Islands (Mallorca), an endemic region of canine leishmaniosis, have reported very few cases of leishmaniosis in Ibizian hounds while concurrently observing that dogs of other breeds had a high incidence of clinical canine leishmaniosis. To further investigate this observation, two populations of dogs from the Balearic Islands were examined for the presence of Leishmania-specific cellular immunity using a delayed type hypersensitivity test (DTH) to leishmanin and for the presence of Leishmania specific humoral immunity using an ELISA. Fifty-six asymptomatic dogs, 31 Ibizian hounds and 25 dogs belonging to other breeds were examined. Seventy-seven percent of the dogs demonstrated a specific immune response against Leishmania, either humoral or cellular. This finding suggests that the infection rate (77%) was higher than previously considered. For Ibizian hounds 81% were DTH positive while only 48% of the other dogs were DTH positive. A statistical association between Ibizian hounds and positive DTH response was found. A specific humoral response was found in 48% of Ibizian hounds and in 56% of the other dogs. No statistical association relative to the Leishmania-specific IgG1 and IgG2 levels were found between the two groups. The Ibizian hound has been reported to be more resistant to Leishmania infection and we found that the Ibizian hound mounts a significant cellular response to infection. Thus, the Ibizian hound may be an interesting canine model for the investigation of protective anti-Leishmania immune response. PMID- 10828511 TI - Serological survey and first finding of Neospora caninum in Taiwan, and the detection of its antibodies in various body fluids of cattle. AB - A serological survey for antibodies against Neospora caninum in cattle, goats and farm dogs in Taiwan was carried out. Sera of 613 cattle from 25 dairy farms, 24 goats from six goat farms and 13 dogs from six dairy cattle farms were tested for antibodies against N. caninum using indirect fluorescence antibody test (IFAT). The same sera were also tested for antibodies against Toxoplasma gondii using latex agglutination test. Of the 613 cattle sera, 44.9% (275/613) were found to have antibodies against N. caninum. Among these 275 positive cattle, 77 also possessed antibodies against T. gondii. Nevertheless, 92 cattle which were negative for N. caninum showed antibodies against T. gondii. Of the 24 goat sera tested, none was found to be positive for N. caninum but 50% (12/24) were positive for T. gondii. Of the 13 farm dogs tested, three were found to possess antibodies against N. caninum, two of which tested negative for T. gondii antibodies. Besides sera, antibodies to N. caninum in cattle could be observed in the milk, vaginal secretion and saliva. However, the order of higher frequency of antibodies detection is in sera, milk, vaginal secretion and saliva. This is the first demonstration of the presence of antibodies to N. caninum in vaginal secretion and saliva of cattle. A 50microm cyst was observed in the brain of one of the 13 prednisolone-treated SPF ICR mice which had been peritoneally inoculated 4 months earlier with the brain homogenate of a serologically N. caninum positive but T. gondii negative cattle. Thus, we have confirmed for the first time the presence of N. caninum in Taiwan and also observed that it is widespread among dairy cattle and farm dogs. PMID- 10828512 TI - Lesions of the enteric nervous system and the possible role of mast cells in the pathogenic mechanisms of migration of schistosome eggs in the small intestine of cattle during Schistosoma bovis infection. AB - The enteric nervous system in the small intestine of cattle during Schistosoma bovis infection was studied by histological stains and immunohistochemical methods. Lesions due to migration of schistosoma eggs were located mainly in the mucous and the submucous layer overlaying the submucous vascular arcades. Granulomas destroyed ganglia, neurons, nerves fibre strands and nerve fibres. Ganglia situated within or near granulomas were infiltrated by mast cells, eosinophils, lymphocytes, globule leukocytes, neutrophils and macrophages. Mast cells were in close contact with degenerating neuronal perikarya. Whereas vasoactive intestinal peptide-like immunoreactivity in the nerves and neurons in the ganglia within and around granulomas was increased, the neurofilament-like immunoreactivity was reduced. Compared to the myenteric and external submucous plexuses, the internal submucous and mucous plexuses were the most damaged. These changes imply reduced functional capacity in the nervous tissue which might cause reduced motility, malabsorption and partly account for the loss of body weight and condition and failure to thrive which occur in schistosomosis. Biotinylated affinity purified swine anti-rabbit and mouse anti-rabbit immunoglobulins reacted nonspecifically with a subset of mast cells. The reaction revealed many mast cells in early forming granulomas and around schistosome egg tracts and infiltration of mast cells into the ganglia of intestinal nerve plexuses. The observation shows a localized, Type I hypersensitivity reaction suggesting for the release of mast cell-derived chemical mediators in the intestinal reaction to trap or evict S. bovis eggs and to cause diarrhoea. PMID- 10828513 TI - Distinguishing morphological features of the third larval stage of ovine Trichostrongylus spp. AB - In this paper we describe a procedure that enables the identification of species of infective third stage (L(3)) Trichostrongylus larvae. Lambs were infected with putatively monospecific infections of three species of Trichostrongylus commonly found in New Zealand (T. axei, T. colubriformis and T. vitrinus) and Teladorsagia (Ostertagia) circumcincta. After recovering L(3) from faecal cultures, the lambs were slaughtered and adult male worms recovered and examined for spicule morphology to verify identification. L(3) were examined for morphological features and measurement of their length. Further L(3) were exsheathed and examined under high power optics to observe posterior morphological features (tubercles). The posterior of T. colubriformis has a three-tubercle structure whereas T. vitrinus has a single tubercle and T. axei none. However, the tails of T. circumcincta also lack tubercles and thus T. axei cannot be readily distinguished from them on this feature. The range of lengths of L(3) of Trichostrongylus spp. (600-858 microm) and T. circumcincta (700-914 microm) were found to overlap considerably. The shape of the anterior end of these two species differs and this in combination with length provides an indication of the proportion of T. axei and T. circumcincta in a culture. A combination of tubercle number, with overall length and anterior morphology of L(3), can be used to differentiate nematode populations of T. axei, T. colubriformis, T. vitrinus and T. circumcincta. PMID- 10828514 TI - Nematode parasitism in adult dairy cows in Belgium. AB - Over a period of 1 year, from November 1997 to October 1998, the abomasa, blood and faecal samples of 121 dairy cows in Belgium were collected and examined for nematode infections. Nematodes were present in the abomasa of 110 animals. Ostertagia was found in all 110, Trichostrongylus was seen in 65 and Haemonchus in 14 abomasa. Overall, 91% of all trichostrongyles recovered were Ostertagia. The geometric mean total number of Ostertagia was 2750, with an average of 74% inhibited early fourth stage larvae (EL4). Between November and February >90% of the Ostertagia worm burden were EL4 stages. The majority of the animals (56%) harboured a low Ostertagia burden (100-5000) and 15% had a high burden (>10,000). Sixty-four percent of the coprocultures were positive and the genera recovered were Ostertagia sp. (100%), Trichostrongylus sp. (42%), Oesophagostomum (32%), Haemonchus sp. (29%) and Cooperia sp. (16%). A seasonal pattern was evident for serum Ostertagia-specific antibodies and for serum pepsinogen concentration, with the highest levels during the summer, and low values during the winter. Dictyocaulus viviparus specific antibodies were detected in the serum of eight (7%) animals. PMID- 10828515 TI - The effectiveness of doramectin pour-on in the control of gastrointestinal nematode infections in cow-calf herds. AB - Two field studies were conducted in the USA to determine the efficacy of a single strategically-timed dose of doramectin pour-on in the control of gastrointestinal nematodosis in beef cow-calf herds and the resultant effects on calf productivity. One study was carried out between May and October 1996 in a spring calving herd at a site located in Idaho (ID) and the other between January and July 1997 in a fall-calving herd at a site located in Mississippi (MS). In each study, cow-calf pairs were randomly allotted by sex of calf to pastures and one of two treatment groups (doramectin pour-on at the recommended dose rate of 500 microg/kg body weight or untreated control). There were four pasture replicates per treatment at each site. Each pasture contained 12 cow-calf pairs at the ID site and 15 cow-calf pairs at the MS site. Treatment was administered to cows and calves on 21 May 1996 (ID) or 23 January 1997 (MS). Following treatment, cow-calf pairs were assigned to their designated pastures where they remained until the calves were weaned 140 (ID) or 168 (MS) days later. Cow and calf fecal egg counts and calf body weights were recorded on treatment day and then at monthly intervals until study termination. Doramectin treatment reduced nematode egg output in cows and calves over the entire grazing season compared to untreated controls and resulted in calf weight gain improvements of 9.8kg (p=0.295) at the ID site and 17.4kg (p=0.0002) at the MS site. PMID- 10828516 TI - Ostertagia ostertagi challenge of calves vaccinated with Haemonchus placei intestinal homogenate. AB - The protective capacity of vaccination with Haemonchus placei whole gut homogenate against challenge with the non-blood-feeding nematode Ostertagia ostertagi was evaluated in calves. Ten helminth-free calves were randomly assigned to two groups. Group 1 received 100microg H. placei intestinal homogenate in the adjuvant 5% dextran sulfate/PBS, while Group 2 received the adjuvant alone. Injections were administered subcutaneously on Days 0 and 28. All calves were challenged with approximately 26,100 O. ostertagi larvae on Day 42. Serum antibody response and counts of nematode eggs per gram of feces (EPG) were determined throughout the study. Calves were necropsied at 5.5 weeks post challenge for recovery of nematodes. Although significant increases were detected in both serum IgG(1) and IgG(2) of Group 1 calves (p<0.05), there was no significant difference in the total number of O. ostertagi recovered from the two groups (p>0.05). Lengths of adult nematodes were not significantly different between groups nor were the numbers of eggs present in adult females recovered from each group significantly different (p>0.05). There were also no significant differences between groups regarding fecal egg counts (p>0.05). Results suggest either: (1) the antigens targeted by the induced antibodies were not present in O. ostertagi; (2) the antigens targeted by the induced antibodies were present, but not essential to O. ostertagi survival; or (3) the antigen was present and essential, but amount of antibody ingested was insufficient to cause damage to the nematode. PMID- 10828517 TI - Evaluation of the effects of nematode parasitism on grazing behaviour, herbage intake and growth in young grazing cattle. AB - Parasitic nematodes, even in the absence of any clinical disease, can cause a reduction in voluntary feed intake in housed ruminants. This trial examined these effects on young cattle grazing pastures. Twenty dairy heifer calves, born in the previous autumn, were blocked according to liveweight and allocated to one of two groups: either untreated or dosed with an IVOMEC((R)) (ivermectin) SR Bolus 10 days prior to turnout on 1 May 1998. The groups grazed separately on two paddocks on predominately ryegrass swards. Liveweights were recorded every 28 days and faecal samples taken for worm egg counts at the same time. In mid-May and mid July, for two 14-day periods, animals were paired within treatment groups and transferred to one of 10 replicate paddocks of similar sward height and herbage mass. Grazing behaviour and herbage intake were measured during these periods. In mid-May, 2 weeks after turnout, treated and untreated animals showed no difference in grazing behaviour or daily intake of grass. By mid-July, 10 weeks after turnout, the untreated heifers spent on average 105min less per day in grazing time and their daily herbage intake was 0.78kg dry matter (DM) per day lower than that of the treated animals. Residual sward height, mass and composition in the trial paddocks reflected these differences. There were no clinical signs of gastrointestinal parasitism in the untreated group up to this time; in July, the mean worm egg count in this group was 120 eggs per gram (e.p.g.) of faeces. Faecal samples from several untreated animals were positive for lungworm larvae by July, mild clinical signs of bronchitis were observed in two of these animals in mid-July. Over the period from turnout until mid-July, the untreated heifers showed a reduction in mean daily liveweight gain of 150g, compared to the treated animals. PMID- 10828518 TI - Inspection of wool lots at sales as a diagnostic test for louse infestation. AB - The accuracy of visual inspection of wool lots for lice as a test for louse infestation was estimated using information provided by 178 woolgrowers in Queensland, Australia. The estimated sensitivity of inspection was 36% (95% confidence interval, 19-58%) and the specificity was 95% (95% CI, 88-98%). Accuracy was influenced by timing, after shearing, of pesticide application for louse control and by class of pesticide last applied after shearing. Visual inspection was less sensitive (29%) if pesticides were applied >3 months after shearing and less sensitive (21%) if an insect growth regulator was the class of pesticide last used after shearing. Based on 36% sensitivity, it was estimated that 16 inspections would have to be conducted to reduce the false negative test rate to <20% in the study population. We suggest that visual inspection of wool lots could be used to efficiently monitor the prevalence of louse infestations in Queensland sheep flocks. Positive inspection results are more likely to represent real louse infestations, rather than a false test result, in flocks grazed in the more extensive regions of Queensland. PMID- 10828519 TI - Detection and quantitation of Ehrlichia risticii genomic DNA in infected horses and snails by real-time PCR. AB - A real-time quantitative PCR using the TaqMan fluorogenic detection system (TaqMan PCR) was established for identification of Ehrlichia risticii, the agent of Potomac horse fever (PHF). The TaqMan PCR identified an 85 base pair section of the 16S rRNA gene by use of a specific fluorogenic probe and two primers. This technique was specific for eight tested E. risticii strains. The TaqMan system identified 10 copies of a cloned section of the 16S rRNA gene of E. risticii. The sensitivity and specificity of the TaqMan PCR were similar to those of conventional nested PCR. The TaqMan PCR was evaluated on horses with infectious colitis and on freshwater stream snails collected from regions with a history of PHF. E. risticii could be detected in 22 of 153 (14.4%) horses with infectious colitis and in 25 of 234 (10.7%) snails in the TaqMan PCR. The same results were obtained in the conventional nested PCR. The Ehrlichia-load was in the range of 10,000-9,000,000 and 35,000-680, 000,000 Ehrlichia equivalents per microg leukocyte DNA and snail DNA, respectively. PMID- 10828520 TI - Evaluation of different methods for the diagnosis of scabies in swine. AB - Scabies in pigs is still very common in many countries and can be detrimental to the productivity of pigs. However, correct diagnosis of the disease can preclude meaningful comparisons of results. Therefore, the objective of the present study was to determine, on 11 pig farms, the prevalence of scabies by determination of the presence of mites in ear scrapings, the dermatitis score, the SI and the detection of specific serum antibodies. For the latter an indirect ELISA technique was performed using a free-living mite as a source of antigen. A second objective was to compare the value of these different diagnostic tests. Four farms were positive for the presence of mites. Our study indicated that the SI of piglets is not reliable as a diagnostic tool for scabies (all values were below the threshold value of 0.4, even on farms that were positive for mites) but on the two farms with the highest prevalence of mites the SI was above the threshold for the finishers. However, the fact that sows from eight of the 11 farms investigated had a SI>0.4 would indicate that for sows either the SI is not very specific, or that a cut-off level of 0.4 is not relevant for this age group. On three of the four infected farms the ADS was higher than the cut-off value of 0.5, and on the fourth farm, where the ADS was only 0.43, individual carcasses with generalised dermatitis (score 2) were present. However, an ADS>0.5 did not always coincide with the presence of mites. On six farms, ODR values were indicative for the presence of Sarcoptes, and on three of these farms this was confirmed with positive ear scrapings. In conclusion, as determined by the detection of mites in pig ears, especially the results from the dermatitis scores seem to be useful in the diagnosis of scabies. The specificity of the other parameters is not sufficient, and therefore, the detection of mites should still be used to confirm scabies on a farm, in combination with other tools. PMID- 10828521 TI - In vitro isolation and characterization of bovine Neospora caninum in Korea. AB - The brains of nine aborted bovine fetuses and two newborn calves born from dams suspected to be infected with Neospora caninum were homogenized and inoculated into Vero cells. All fetuses and calves were from cows determined as seropositive to N. caninum by an IFA test. Sera and thoracic fluids of all fetuses and calves also revealed high antibody titer to N. caninum by IFAT ranging from 1:800 to 1:3200. N. caninum was isolated from the brains of one aborted fetus and one newborn calf when the brain homogenates were grown continuously in Vero cell culture. N. caninum tachyzoites, giemsa-positive, were first observed on Days 45 and 56 postinoculation in the newborn calf and the aborted fetus, respectively. The isolates (KBA-1 and KBA-2) were morphologically and ultrastructurally similar to previously published Neospora isolates. The isolated parasites were confirmed as N. caninum by means of the antigenic reactivities, immunostaining, PCR and southern blotting, and electron microscopy. PMID- 10828522 TI - Camel trypanosomosis in the Canary Islands: assessment of seroprevalence and infection rates using the card agglutination test (CATT/T. evansi) and parasite detection tests. AB - Trypanosomosis due to Trypanosoma evansi (surra) is a major enzootic disease of the dromedary camel. Thus, the purpose of the present study was to assess seroprevalence and infection rates in the Canary Islands using antibody(-card agglutination test-CATT/T. evansi) and parasite detection tests (micro Haematocrit Centrifugation technique, Giemsa stained blood smears, microscopic examination of lymph node aspirates and mouse inoculation). PCV was also determined. 745 dromedary camels (483 females and 262 males) were examined. Trypanosomes were detected in seven animals. 36 animals yielded CATT positive results while 709 animals were negative. All parasitologically positive animals were also CATT positive. Results showed a good correlation between CATT positive and low PCV and a higher seroprevalence in older animals. Trypanocidal drugs have not been registered in Spain and, consequently, if vigilance is not exercised the prevalence could be increased in the future. PMID- 10828523 TI - No evidence for cholinergic problems in apolipoprotein E knockout and apolipoprotein E4 transgenic mice. AB - The varepsilon4 allele of the apolipoprotein E gene constitutes the major genetic risk factor to develop Alzheimer's disease. If and how this protein contributes to the pathological cascade of Alzheimer's disease is not known. The varepsilon4 allele particularly affects the cholinergic defect, which is one of the most consistent neurotransmitter problems in an Alzheimer's disease brain. We have analysed several parameters of the cholinergic system in brain of apolipoprotein E knockout mice as well as in transgenic mice overexpressing human apolipoprotein E4. We analysed the distribution of cholinergic fibers, the number and morphology of cholinergic neurons and the enzymatic activity of acetylcholinesterase and choline acetyltransferase in different brain regions. Finally, we analysed the distribution and the binding parameters of [3H]hemicholinium-3, a specific marker for the high affinity choline transporter in different brain sections and regions. This extensive effort failed to show any consistent difference in the cholinergic parameters studied, in either the apolipoprotein E4 transgenic mice or in the apolipoprotein E knockout mice, compared to age-matched non-transgenic mice. We conclude that the apolipoprotein E4 is not deleterious per se for the cholinergic system in mouse brain. PMID- 10828524 TI - Reversal of presynaptic deficits of apolipoprotein E-deficient mice in human apolipoprotein E transgenic mice. AB - Apolipoprotein E genotype is an important risk factor of Alzheimer's disease, which is associated with the degeneration of distinct brain neuronal systems. In the present study we employed apolipoprotein E-deficient mice and human apolipoprotein E3 and apolipoprotein E4 transgenic mice on a null mouse apolipoprotein E background, to examine the extent to which distinct brain neuronal systems are affected by apolipoprotein E and the isoform specificity of this effect. This was pursued by histological and autoradiographic measurements utilizing neuron specific presynaptic markers. The results thus obtained revealed significant reductions in the levels of brain cholinergic and noradrenergic nerve terminals in young apolipoprotein E-deficient mice and no changes in brain dopaminergic nerve terminals. These cholinergic and noradrenergic presynaptic derangements were ameliorated similarly in human apolipoprotein E3 and apolipoprotein E4 transgenic mice. In the case of the cholinergic system, this resulted in complete reversal of the presynaptic deficits, whereas in the case of the noradrenergic neurons the amelioration was partial. These findings suggest that brain cholinergic and noradrenergic neurons are markedly more dependent on brain apolipoprotein E than brain dopaminergic neurons and that the isoform specificity of these effects is not apparent at a young age under non-challenged conditions. PMID- 10828525 TI - Modulation of the ischemia-induced taurine release by adenosine receptors in the developing and adult mouse hippocampus. AB - The release of the inhibitory amino acid taurine is markedly enhanced under ischemic conditions in both adult and developing hippocampus, together with a pronounced increase in the release of excitatory amino acids and the neuromodulator adenosine. We studied the effects of adenosine receptor agonists and antagonists as well as adenosine transport inhibitors on hippocampal [(3)H]taurine release in normoxia and ischemia, using a superfusion system. Under standard conditions the adenosine A(1) receptor agonists N(6)-cyclohexyladenosine and R(-)N(6)-(2-phenylisopropyl)adenosine potentiated basal taurine release in developing mice and depressed the release in adults in a receptor-mediated manner. Adenosine A(2) receptor compounds had only minor effects on the basal release and the K(+)-stimulated release was not affected by these drugs. The adenosine uptake inhibitor dipyridamole enhanced basal taurine release in the developing hippocampus and reduced it in the adult. In ischemia the adenosine compounds had no marked effects on taurine release in immature animals, whereas A(1) receptor activation was still able to evoke taurine release in adults by a receptor-mediated mechanism. The results show that the basal release of taurine is modulated by A(1) receptors in both mature and immature hippocampus, whereas in ischemia these receptors potentiate taurine release only in adults. The elevated taurine levels together with the depression of excitatory amino acid release by adenosine receptor activation could be beneficial under ischemic conditions, protecting neural cells against excitotoxicity and hyperexcitation. PMID- 10828526 TI - Oxidative stress is involved in seizure-induced neurodegeneration in the kindling model of epilepsy. AB - Reactive oxygen species have been implicated in the development of seizures under pathological conditions and linked to seizure-induced neurodegeneration. There has been little direct evidence, however, of free radical production resulting from seizures. Using amygdala-kindled rats, we have examined the generation of reactive oxygen species following seizures, and their possible contribution to seizure development and seizure-induced neuronal loss. The concentrations of two products of free radical-induced lipid peroxidation, malonaldehyde and 4-hydroxy 2(E)-nonenal, were measured using colorimetric assays. Lipid peroxidation was increased in both hemispheres of kindled rats as compared to sham-operated controls. Cell death was also significantly increased in all hippocampal areas. Antioxidants (vitamin E and glutathione) prevented the rise in lipid peroxides and hippocampal neuronal death during kindling, but did not arrest the development of seizures.Thus, epileptiform activity can result in free radical production which may be one of the factors leading to cell death. PMID- 10828527 TI - Interruption of supramammillohippocampal afferents prevents the genesis and spread of limbic seizures in the hippocampus via a disinhibition mechanism. AB - In this study we describe the preventive effect of interruption of the supramammillohippocampal afferents on the Fos expression in the forebrain and epileptic discharges in the hippocampal electroencephalogram in rat model of kainic acid-induced limbic seizure. Little was known about the contribution of different degrees of neural activity of hippocampal principal cells to the genesis and spread of limbic seizures in the forebrain structures. Following kainic acid injection to the amygdala with or without concurrent injection of muscimol to the supramammillary nucleus, behavioral changes and electroencephalograms were observed in freely moving rats. The animals were processed for Fos immunocytochemical analysis at several time points. The latest expression of Fos at 2h was seen in hippocampal CA1-CA3, ventrolateral thalamic nuclei and mediodorsal caudate putamen, while the early Fos expression at 0.5h was seen in the piriform, entorhinal and other cortices, the thalamic midline nuclei and hypothalamic nuclei. Muscimol injection to the supramammillary nucleus prevented Fos expression in the CA1-CA3 region and reduced that in the forebrain regions with the latest Fos expression, but did not affect Fos expression in other forebrain regions with early Fos expression. This treatment also eliminated epileptic discharges and attenuated all waves in hippocampus. These findings indicate that an acute interruption of the facilitatory hypothalamic afferents by intrasupramammillary injection of muscimol may cause the inactivation of the disinhibition mechanism for hippocampal throughput at the dentate gyrus, resulting in the blockade of the genesis and spread of limbic seizures in the hippocampus. PMID- 10828528 TI - Similar increases in extracellular lactic acid in the limbic system during epileptic and/or olfactory stimulation. AB - Previous studies have shown that physiological stimulation of brain activity increases anaerobic glucose consumption, both in humans and in experimental animals. To investigate this phenomenon further, we measured extracellular lactate levels within different rat brain regions, using microdialysis. Experiments were performed comparing the effects of natural, physiological olfactory stimulation of the limbic system with experimental limbic seizures. Olfactory stimulation was carried out by using different odors (i.e. both conventional odors: 2-isobutyl-3-methoxypyrazine, green pepper essence; thymol; and 2-sec-butylthiazoline, a sexual pheromone). Limbic seizures were either induced by systemic injection of pilocarpine (200-400 mg/kg) or focally elicited by microinfusions of chemoconvulsants (bicuculline 118 pmol and cychlothiazide 1.2 nmol) within the anterior piriform cortex. Seizures induced by systemic pilocarpine tripled lactic acid within the hippocampus, whereas limbic seizures elicited by focal microinfusion of chemoconvulsants within the piriform cortex produced a less pronounced increase in extracellular lactic acid. Increases in extracellular lactate occurring during olfactory stimulation with the sexual pheromone (three times the baseline levels) were non-significantly different from those occurring after systemic pilocarpine. Increases in lactic acid following natural olfactory stimulation were abolished both by olfactory bulbectomy and by the focal microinfusion of tetrodotoxin, while they were significantly attenuated by the local application of the N-methyl-D-aspartate antagonist AP-5. Increases in hippocampal lactate induced by short-lasting stimuli (olfactory stimulation or microinfusion of subthreshold doses of chemoconvulsants, bicuculline 30 pmol) were reproducible after a short delay (1 h) and cumulated when applied sequentially. In contrast, limbic status epilepticus led to a long-lasting refractoriness to additional lactate-raising stimuli and there was no further increase in lactate levels when the olfactory stimulation was produced during status epilepticus. Increases in lactic acid following olfactory stimulation occurred with site specificity in the rhinencephalon (hippocampus, piriform and entorhinal cortex) but not in the dorsal striatum. Site specificity crucially relied on the quality of the stimulus. For instance, other natural stimuli (i.e. tail pinch) produced a similar increase in extracellular lactate in all brain areas under investigation. The major conclusion of this work is that the presentation of an odor known to be a rat pheromone results in lactate production as great as that induced by the systemic convulsant pylocarpine (maximum: 2.286+/ 0.195 mM and 1.803+/-0.108 mM, respectively). This supports the notion that the great magnitude of lactate production known to accompany seizures can result from the intensified neural activity per se ("aerobic gycolysis"), not merely from local anoxia or other pathological changes. PMID- 10828529 TI - The effects of ibotenic acid lesions of the medial and lateral prefrontal cortex on latent inhibition, prepulse inhibition and amphetamine-induced hyperlocomotion. AB - Hypofunction of prefrontal cortical regions, such as dorsolateral and orbital regions, has been suggested to contribute to the symptomatology of schizophrenia. In the rat, the medial and the lateral prefrontal cortices are considered as homologs of the primate dorsolateral and orbital prefrontal cortices, respectively. The present study investigated in rats the effects of lesions of the medial and lateral prefrontal cortices on latent inhibition, prepulse inhibition and amphetamine-induced activity. These paradigms are known to be modulated by the mesolimbic dopaminergic system, a system that has been suggested to be involved in the symptomatology of schizophrenia. Latent inhibition and prepulse inhibition are disrupted in schizophrenic patients as well as in rats treated with amphetamine. Amphetamine-induced activity was tested under dim light (low stress) and bright light (high stress) because stressful situations selectively increase mesocortical dopamine activity. Lateral prefrontal cortex lesioned animals did not differ in their behavior from control animals in any of the paradigms used in this study. Medial prefrontal cortex lesions did not affect latent inhibition but increased prepulse inhibition. In the amphetamine-induced activity experiment, prior to drug administration, open field locomotion was reduced under bright illumination for all lesion groups. After amphetamine administration, medial prefrontal cortex lesions attenuated the hyperlocomotor effect of the drug under the dim light condition and potentiated it under the bright light condition. The results indicate that medial and lateral prefrontal cortex can be functionally differentiated by their involvement in the modulation of behavior requiring mesocorticolimbic dopamine activation. The results in amphetamine induced activity suggest that the behavioral outcomes associated with medial prefrontal cortex depend on the background (stress) against which the evaluation is made. The results also support the notion that prepulse inhibition may be a better model than latent inhibition of the symptoms of schizophrenia associated with dysfunctional prefrontal activity. PMID- 10828530 TI - Differential involvement of dopamine in the shell and core of the nucleus accumbens in the expression of latent inhibition to an aversively conditioned stimulus. AB - Latent inhibition, the process whereby pre-exposure to a conditioned stimulus without consequence impairs subsequent learning of an association between the conditioned stimulus and an unconditioned stimulus, is reportedly disrupted in both amphetamine-treated rats and in acute schizophrenics. This has led to the suggestion that disruptions in latent inhibition model some of the cognitive impairments associated with hyperactive dopamine transmission as it is expressed in schizophrenic patients. Specifically, fluctuations in dopamine neurotransmission within the nucleus accumbens have been implicated in the mediation of latent inhibition; however, it has not been established whether these dopamine-mediated effects occur in the shell or core subregion of the nucleus. In the present study, 48h after conditioned stimulus-pre-exposed and non pre-exposed animals experienced 10 pairings of tone and footshock, we measured extracellular levels of dopamine in the shell and core during the expression of latent inhibition to an aversively conditioned tone using in vivo microdialysis. Our results show that pre-exposure to the tone eliminated the conditioned release of dopamine in the shell of the nucleus accumbens and resulted in an attenuated conditioned freezing response to the tone conditioned stimulus. In contrast, dopamine release in the core was not affected by pre-exposure to the tone. These data suggest that it is specifically the shell of the nucleus accumbens in which alterations of dopaminergic tone, whether pharmacologically induced in rodents or the result of disease in humans, may act to disrupt latent inhibition. PMID- 10828531 TI - The central nucleus of the amygdala projection to dopamine subpopulations in primates. AB - The dopamine system plays a major role in responses to potentially rewarding stimuli. An important input to the dopamine neurons is derived from the central nucleus of the amygdala. The central nucleus is a complex structure consisting of several subdivisions with distinct histochemical, morphologic, and connectional features. The central nucleus subdivisions are therefore likely to have specific inputs to the dopamine neurons. The midbrain dopamine cells are divided into dorsal and ventral subpopulations. We determined the organization of inputs from the central nucleus subdivisions to the dopamine subpopulations in monkeys. The dorsal tier neurons receive relatively greater central nucleus input compared to the ventral tier. Within the ventral tier, the central nucleus projects to the densocellular region, but not the cell columns. Furthermore, the midbrain subpopulations receive a differential projection from specific central nucleus subterritories. The medial subdivision of the central nucleus has the greatest input to the dopamine system, and projects throughout the dorsal tier and densocellular regions. This indicates that the medial subdivision influences not only the ventral striatum but also more dorsal striatal areas, through its inputs to these dopamine subpopulations. In contrast, the capsular subdivision of the lateral central nucleus and the amygdalostriatal area project preferentially to the dorsal tier, which selectively modulates the ventral striatum and cortex. The central core of the lateral central nucleus is unique in its restricted projection to the lateral substantia nigra in the region of the nigrotectal pathway. Taken as a whole, the central nucleus-nigral pathway provides a route for affectively significant stimuli to modulate the DA system, influencing the initiation of behavioral responses. PMID- 10828532 TI - Targeted disruption of the dopamine D(2) and D(3) receptor genes leads to different alterations in the expression of striatal calbindin-D(28k). AB - The present study used mice deficient for dopamine D(2) and D(3) receptors to test whether the expression of these two members of the D(2) class of receptors is essential for the normal expression of three markers that characterize the neurochemical differentiation of the striatum: the calcium-binding protein calbindin, tyrosine hydroxylase and acetylcholinesterase. Results from these experiments revealed that the expression of striatal tyrosine hydroxylase (the rate-limiting enzyme of dopamine synthesis) and acetylcholinesterase is unaffected even by the combined knockout of D(2) and D(3) receptors. However, D(2) and D(3) receptor knockouts differently affect the striatal expression of calbindin-D(28k) immunoreactivity. Prominent changes in the cellular distribution of calbindin are detected in striatal neurons of D(2) mutant mice. Whereas calbindin immunolabeling of wild-type neurons is prominent in the nuclei and the cytoplasm of medium spiny neurons, in D(2) mutant mice, calbindin immunoreactivity is concentrated exclusively in the cytoplasmic rim of these neurons. Such changes in the cellular distribution of calbindin expression are not detected in mice lacking D(3) receptors. In these mutants, however, a lesser density of calbindin-immunoreactive neuropil is detected in the ventral portions of the striatum, i.e. in regions in which D(3) receptors are thought to be expressed at highest levels. Mice lacking both D(2) and D(3) receptors show both phenotypes. The altered cellular distribution of calbindin in D(2) mutants is likely to have functional consequences for some of the Ca(2+)-mediated cellular functions. The topography of the decreased density of striatal calbindin immunorectivity in D(3) mutants suggests a role for D(3) receptors in supporting the expression of striatal calbindin. The observation that mice lacking both D(2) and D(3) receptors show a combination of the D(2) and D(3) mutant phenotypes indicates that each of the different phenotypes detected in the single mutants is indeed related to the lack of the two different D(2)-like receptor subtypes. PMID- 10828533 TI - The pattern of neurodegeneration in Huntington's disease: a comparative study of cannabinoid, dopamine, adenosine and GABA(A) receptor alterations in the human basal ganglia in Huntington's disease. AB - In order to investigate the sequence and pattern of neurodegeneration in Huntington's disease, the distribution and density of cannabinoid CB(1), dopamine D(1) and D(2), adenosine A(2a) and GABA(A) receptor changes were studied in the basal ganglia in early (grade 0), intermediate (grades 1, 2) and advanced (grade 3) neuropathological grades of Huntington's disease. The results showed a sequential pattern of receptor changes in the basal ganglia with increasing neuropathological grades of Huntington's disease. First, the very early stages of the disease (grade 0) were characterized by a major loss of cannabinoid CB(1), dopamine D(2) and adenosine A(2a) receptor binding in the caudate nucleus, putamen and globus pallidus externus and an increase in GABA(A) receptor binding in the globus pallidus externus. Second, intermediate neuropathological grades (grades 1, 2) showed a further marked decrease of CB(1) receptor binding in the caudate nucleus and putamen; this was associated with a loss of D(1) receptors in the caudate nucleus and putamen and a loss of both CB(1) and D(1) receptors in the substantia nigra. Finally, advanced grades of Huntington's disease showed an almost total loss of CB(1) receptors and the further depletion of D(1) receptors in the caudate nucleus, putamen and globus pallidus internus, and an increase in GABA(A) receptor binding in the globus pallidus internus. These findings suggest that there is a sequential but overlapping pattern of neurodegeneration of GABAergic striatal efferent projection neurons in increasing neuropathological grades of Huntington's disease. First, GABA/enkephalin striatopallidal neurons projecting to the globus pallidus externus are affected in the very early grades of the disease. Second, GABA/substance P striatonigral neurons projecting to the substantia nigra are involved at intermediate neuropathological grades. Finally, GABA/substance P striatopallidal neurons projecting to the globus pallidus internus are affected in the late grades of the disease. In addition, the finding that cannabinoid receptors are dramatically reduced in all regions of the basal ganglia in advance of other receptor changes in Huntington's disease suggests a possible role for cannabinoids in the progression of neurodegeneration in Huntington's disease. PMID- 10828534 TI - Major strain differences in response to chronic systemic administration of the mitochondrial toxin 3-nitropropionic acid in rats: implications for neuroprotection studies. AB - Chronic systemic treatment with 3-nitropropionic acid in rats produces persistent dystonia and bradykinesia, and striatal lesions reminiscent of Huntington's disease. However, the interpretation of results obtained with this model are complicated by a heterogeneous distribution of the response to a given toxic dose of 3-nitropropionic acid: approximately half of the animals develop selective striatal lesions, which in certain cases are associated with extrastriatal lesions, and the other half are apparently spared. Thus, the chronic 3 nitropropionic acid lesion model can be difficult for neuroprotection studies in which a consistent response to neurotoxic treatment is prerequisite. We hypothesized that some of the variability in the model was related to the use of Sprague-Dawley rats, since inter-individual variability in response to various stressful conditions has been described previously in this rat strain. We therefore compared 3-nitropropionic acid toxicity in rat strains known to be highly (Fisher 344) or poorly (Lewis) responsive to stress and compared the distribution of responses to that of Sprague-Dawley rats. In a protocol of intraperitoneal injection, toxicity of 3-nitropropionic acid was highest in Fisher rats, intermediate in Sprague-Dawley rats and lowest in Lewis rats. In addition, survival curves showed a more heterogeneous response to 3 nitropropionic acid toxicity in Sprague-Dawley rats than that observed in Lewis and Fisher rats. These differences between Sprague-Dawley and Lewis rats were confirmed in a protocol of subcutaneous 3-nitropropionic acid intoxication using osmotic minipumps, where doses up to 36-45mg/kg per day for five days were necessary to induce striatal lesions in Lewis rats as compared to 12-14mg/kg per day for five days in Sprague-Dawley rats. The selectivity of the striatum to lesions, and homogeneous progression of symptoms and neurodegeneration, were more consistently observed in Lewis as compared to Sprague-Dawley rats. These results suggest that vulnerability to 3-nitropropionic acid may depend on genetic factors, which could also influence the physiological response to stress. The present findings also establish an improved model of progressive striatal degeneration in the rat adapted for the testing of new neuroprotective strategies. PMID- 10828535 TI - Cellular and mitochondrial changes in glutamate-induced HT4 neuronal cell death. AB - Elevated levels of extracellular glutamate are neurotoxic. The cytotoxic property of extracellular glutamate is known to mediate two primary mechanisms, excitotoxicity and excitotoxicity-independent processes. The excitotoxicity independent pathway was investigated in the current study in a mouse hippocampal derived HT4 cell line. Exposure of HT4 cells to glutamate for 12h induced loss of cell viability preceded by rapid loss of intracellular reduced glutathione followed by accumulation of intracellular reactive oxygen species, elevation of intracellular Ca(2+), progressive loss of mitochondrial membrane potential swelling and loss of mitochondrial outer membrane integrity. Glutamate-induced loss of DNA integrity has been detected. The antioxidants alpha-tocopherol and trolox, mitochondrial calcium uniporter inhibitor Ruthenium Red and protein synthesis inhibitor cycloheximide all showed protection against glutamate-induced toxicity. None of the protective agents except for alpha-tocopherol controlled the glutamate-induced reactive oxygen species build-up. However, these cell death regulators prevented the glutamate-induced mitochondrial damage and regulated glutamate-induced increase in intracellular Ca(2+). Carbonyl cyanide p trifluoromethoxyphenyl-hydrazone, a mitochondrial uncoupler, partially protected against glutamate-induced cell death and mitochondrial damage, while the mitochondrial ribosomal inhibitor chloramphenicol and extracellular Ca(2+) chelator ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid did not protect the cells against glutamate treatment. The results of this study demonstrated that mitochondrial dysfunction was a key event in the excitotoxicity independent component of neuronal cell death. Reactive oxygen species accumulation and glutathione depletion were prominent in glutamate-treated cells; however, these events were not direct mediators of cell death. PMID- 10828536 TI - Contrasting effects of noradrenergic beta-receptor blockade within the medial septal area on forebrain electroencephalographic and behavioral activity state in anesthetized and unanesthetized rat. AB - The locus coeruleus-noradrenergic system participates in the modulation of behavioral state. Previous studies demonstrated that beta-receptors located within the general region encompassing the medial septum/vertical limb of the diagonal band of Broca (medial septal area) exert arousal-enhancing actions in both anesthetized and unanesthetized animals. These studies also demonstrated that, under conditions of limited locus coeruleus discharge rates, blockade of beta-receptors within this region decreased forebrain electroencephalographic indices of arousal. The current studies assess the extent to which medial septal area beta-receptors contribute to the maintenance of electroencephalographic and/or behavioral indices of arousal, under conditions associated with elevated locus coeruleus discharge rates. In the halothane-anesthetized rat, bilateral, but not unilateral, blockade of beta-receptors within this area prevented forebrain (cortical and hippocampal) electroencephalographic activation elicited by activation of locus coeruleus neurons. Placement of beta-antagonist immediately adjacent to the medial septal area had no effect on locus coeruleus dependent cortical and hippocampal electroencephalographic activation. In contrast, in unanesthetized rat, bilateral pretreatment of the medial septal area did not alter either electroencephalographic or behavioral measures in animals tested in an arousal-enhancing, brightly-lit novel environment, which has been demonstrated to elicit an activation of the locus coeruleus-noradrenergic system. The results obtained in anesthetized animals are consistent with previous studies demonstrating potent modulatory actions of noradrenergic systems on actions of general anesthetics, and suggest that beta-receptors may be an appropriate target for pharmacological adjuncts to general anesthetics. In contrast to that observed in anesthetized animals, medial septal beta-receptors alone do not contribute significantly to the maintenance of an activated forebrain in unanesthetized animals. It is presumed that actions of other noradrenergic receptors and/or other neurotransmitter systems located within or outside the medial septal area make the arousal-modulating actions of medial septal area beta-receptors redundant, in the unanesthetized, alert animal. PMID- 10828537 TI - Synaptic contacts between serotonergic and cholinergic neurons in the rat dorsal raphe nucleus and laterodorsal tegmental nucleus. AB - We examined synaptic connectivity between cholinergic and serotonergic neurons in the dorsal raphe nucleus and the laterodorsal tegmental nucleus of the rat. To this purpose we employed two variations (the combination of pre-embedding immunogold-silver intensification with avidin-biotin-peroxidase complex technique and the combination of avidin-biotin-peroxidase/3, 3'-diaminobenzidine/silver gold intensification with avidin-biotin-peroxidase/3,3'-diaminobenzidine reaction) of a double pre-embedding immunoelectron procedure, using primary antibodies against vesicular acetylcholine transporter and serotonin. At the light-microscopic level, serotonin-like immunoreactive neurons in the dorsal raphe nucleus appeared as reddish black and vesicular acetylcholine transporter like immunoreactive axon terminals were brown colored using a combination of pre embedding immunogold-silver technique and avidin-biotin-peroxidase complex technique. Serotonin-like immunoreactive fibers projected to the laterodorsal tegmental nucleus. At the electron microscopy level, with both methods we observed in the dorsal raphe nucleus vesicular acetylcholine transporter immunopositive axon terminals in synaptic contact with serotonin-like immunoreactive dendrites and, to a lesser degree, with serotonin-like immunoreactive cell bodies. These synapses usually were of the symmetrical type. Occasionally we noted, next to vesicular acetylcholine transporter-immunopositive axon terminals, also immunonegative terminals synapsing with the serotonin-like immunoreactive dendrites. In the laterodorsal tegmental nucleus we found serotonin-like immunoreactive axon terminals and immunonegative terminals forming synapses with vesicular acetylcholine transporter-immunoreactive dendrites. Most synapses formed by the serotonin-like immunopositive terminals were of the asymmetrical type. Our results suggest that serotonergic neurons in the dorsal raphe nucleus and cholinergic neurons in the laterodorsal tegmental nucleus may reciprocally influence each other by means of synaptic connectivity. Such connectivity may serve to regulate pain sensation, or be involved in the regulation of the sleeping-waking cycle. PMID- 10828538 TI - Cerebellar output exerts spatially organized influence on neural responses in the rat superior colliculus. AB - The deep cerebellar nuclei project to largely segregated target regions in the contralateral superior colliculus. Single-unit recordings have previously shown that nuclear inactivation normally suppresses spontaneously active collicular target neurons. However, facilitation of activity has also been found in a proportion of collicular units. In the present study we tested the hypothesis that the type of effect is related to the cerebellotectal topography. We recorded simultaneously in the deep cerebellar nuclei and superior colliculus of 53 anaesthetized rats. GABA microinjections produced a complete, reversible, arrest of activity in the deep cerebellar nuclei. We investigated the effect of this inactivation on 292 sensory and non-sensory cells in the collicular intermediate and deep layers. Of these, 29% showed a reduced response to their preferred sensory stimulus or decreased their spontaneous firing rate in the case of non sensory cells. However, 15% increased their sensory responsiveness and/or spontaneous firing rate following cerebellar inactivation. No effect was seen in the remaining 56% of cells. The distribution of these different effects was highly significantly related to the topography of the cerebellotectal terminal fields. Thus, 68% of the suppressive effects were obtained from cells lying in the terminal fields of the deep cerebellar nucleus inactivated. Conversely, 86% of the excitatory effects and 66% of the cells showing no effect were obtained from cells falling outside the terminal field. The results support the view that the superior colliculus is an important site for the functional integration of primary sensory information, not only with cortical and basal ganglia afferents, but also with cerebellar information. The contrasting physiological responses observed within the terminal cerebellotectal topography appear to map closely on to the known distribution of the cells of origin of the two major descending output pathways of the superior colliculus and are possibly mediated by intrinsic inhibitory connections within its intermediate and deep layers. These results provide evidence for a neural architecture in the superior colliculus whose function is the selection of appropriate actions in response to novel stimuli and the suppression of competing motor programmes. PMID- 10828539 TI - Anterograde axonal transport of glial cell line-derived neurotrophic factor and its receptors in rat hypoglossal nerve. AB - Glial cell line-derived neurotrophic factor is one of the most potent motoneuron survival factors yet identified. Although retrograde transport of trophic factors to the cell body is thought to be an important process in motoneuron survival, the transport pathways that lead to interaction of glial cell line-derived neurotrophic factor with its receptors is not known. We have used a double ligated hypoglossal nerve preparation to investigate transport of endogenous glial cell line-derived neurotrophic factor and its receptors, glial cell line derived neurotrophic factor family receptor alpha1 and receptor re-arranged during transfection. Glial cell line-derived neurotrophic factor was found to accumulate at the distal ligature, indicating retrograde transport and consistent with its motoneuron survival effects. In addition, we observed accumulation of glial cell line-derived neurotrophic factor and its receptors at the proximal ligature, indicating anterograde transport. This finding is not predicted by neurotrophic theory. Staining for glial cell line-derived neurotrophic factor in the motor axons was punctate, suggesting involvement of transport vesicles. Results obtained using immunohistochemistry and reverse transcription-polymerase chain reaction provide evidence for the synthesis of glial cell line-derived neurotrophic factor and glial cell line-derived neurotrophic factor family receptor alpha1 in Schwann cells and glial cell line-derived neurotrophic factor family receptor alpha1 and receptor re-arranged during transfection in motoneuron cell bodies. When the motor axons were separated from the cell body by avulsion, glial cell line-derived neurotrophic factor remained in the vicinity of the Schwann cells and did not accumulate at the proximal ligature. Our results indicate anterograde transport of Schwann cell-derived glial cell line-derived neurotrophic factor, which is dependent on binding to its cell body-derived receptors. These findings suggest a mechanism for collection of glial cell line derived neurotrophic factor from multiple Schwann cells which surround motor axons. We propose that in addition to its role in motoneuron survival, glial cell line-derived neurotrophic factor may also modulate local neuronal effects in distal regions of the nerve. PMID- 10828540 TI - Facial motor neuron regeneration induces a unique spatial and temporal pattern of myristoylated alanine-rich C kinase substrate expression. AB - We have previously shown that the myristoylated alanine-rich C kinase substrate, a primary protein kinase C substrate in brain that binds and cross-links filamentous actin, is enriched in neuronal growth cones and is developmentally regulated in brain. Here we examined myristoylated alanine-rich C kinase substrate expression in the facial motor nucleus during axonal regeneration following facial nerve axotomy or facial nerve resection lesions, which impede regeneration, or following motor neuron degeneration induced by the retrograde neurotoxin ricin. For comparative purposes, the protein kinase C substrates myristoylated alanine-rich C kinase substrate-like protein and growth-associated protein-43 were examined in parallel. Myristoylated alanine-rich C kinase substrate messenger RNA exhibited a robust increase in both neurons and non neuronal cells in the facial motor nucleus beginning four days after axotomy, peaked at seven days (2.5-fold), and declined back to baseline levels by 40 days. Myristoylated alanine-rich C kinase substrate protein similarly exhibited a twofold elevation in the facial motor nucleus determined four and 14 days post axotomy. Following nerve resection, myristoylated alanine-rich C kinase substrate messenger RNA levels increased at seven days and returned to baseline levels by 40 days. Unlike myristoylated alanine-rich C kinase substrate messenger RNA, myristoylated alanine-rich C kinase substrate-like messenger RNA levels did not increase in the facial motor nucleus at any time point following nerve axotomy or resection, whereas growth-associated protein-43 messenger RNA exhibited a rapid (one day) and prolonged (40 days) elevation in facial motor nucleus neurons following either nerve axotomy or resection. Ricin-induced degeneration of facial motor neurons elevated myristoylated alanine-rich C kinase substrate and myristoylated alanine-rich C kinase substrate-like messenger RNAs in both microglia (lectin-positive) and astrocytes (glial fibrillary acidic protein positive).Collectively, these data demonstrate that myristoylated alanine-rich C kinase substrate exhibits a unique expression profile in the facial motor nucleus following facial nerve lesions, and it is proposed that myristoylated alanine rich C kinase substrate may serve to mediate actin-membrane cytoskeletal plasticity in both neurons and glial cells in response to protein kinaseC mediated signaling during nerve regeneration and degeneration. PMID- 10828541 TI - Expression of multiple functional chemokine receptors and monocyte chemoattractant protein-1 in human neurons. AB - Functional chemokine receptors and chemokines are expressed by glial cells within the CNS, though relatively little is known about the patterns of neuronal chemokine receptor expression and function. We developed monoclonal antibodies to the CCR1, CCR2, CCR3, CCR6, CXCR2, CXCR3 and CXCR4 chemokine receptors to study their expression in human fetal neurons cultured from brain tissue as well as the clonally derived NT2.N human neuronal cell line (NTera 2/cl.D1). Specific monoclonal antibody labeling demonstrated expression of CCR2, CXCR2, CXCR3 and CXCR4 on neurons from both sources. Co-labeling studies revealed strong expression of CXCR3 and CXCR4 on both dendritic and axonal processes, with a weaker expression of CXCR2 and CCR2. Reverse transcriptase-polymerase chain reaction analysis of pure NT2.N neurons confirmed RNA expression for CCR2, CXCR2, CXCR3 and CXCR4. No changes in the neuronal labeling pattern of chemokine receptor expression were noted when NT2.N neurons were grown on a supporting layer of astrocytes, again consistent with similar patterns seen in primary human fetal brain cultures. Analysis of single-cell calcium transients revealed a robust response to stromal derived factor-1alpha (CXCR4) and melanocyte growth stimulating activity (CXCR2), and variable response to monocyte chemoattractant protein-1 (CCR2) or interferon-gamma inducible protein-10 (CXCR3). Finally, we detected the release of monocyte chemoattractant protein-1 from pure cultures of NT2.N neurons, but not undifferentiated NT2 cells. These data indicate that individual neurons may not only co-express multiple functional chemokine receptors, but also that neurons themselves produce chemokines which may influence cellular function within the central nervous system. PMID- 10828542 TI - Uptake and metabolism of glutamate at non-synaptic regions of crayfish central nerve fibers: implications for axon-glia signaling. AB - In crayfish and squid giant nerve fibers, glutamate appears to be an axon-glia signaling agent. We have investigated glutamate transport and metabolism by crayfish central nerve fibers in order to identify possible mechanisms by which glutamate could subserve this non-synaptic signaling function. Accumulation of radiolabeled L-glutamate by desheathed cephalothoracic nerve bundles was temperature and Na(+) dependent, linear with time for at least 8h and saturable at about 0.5-1mM L-glutamate. Most accumulated radiotracer was associated with the periaxonal glial sheath and remained as glutamate. Compounds known to block glutamate transport in invertebrate peripheral nerves or mammalian brain slices or cell cultures were also effective on crayfish central nerve fibers. Tissue radiotracer levels were only 3% of control levels when 1mM p chloromercuriphenylsulfonate was present, and 13%, 20%, 26%, 38% and 42% of control levels, respectively, when L-cysteate, L-cysteine sulfinate, L-aspartate, D-aspartate or DL-threo-beta-hydroxyaspartate was present. L-Glutamine, GABA, N methyl-DL-aspartate, alpha-aminoadipate and D-glutamate were without inhibitory effect on tissue tracer accumulation. Radiolabeled D-aspartate was an equivalent non-metabolized substitute for radiolabeled L-glutamate. D-Aspartate, p chloromercuriphenylsulfonate and GABA had comparable effects on isolated medial giant nerve fibers.These studies indicate that L-glutamate is taken up primarily by the periaxonal glia of crayfish central nerve fibers by a low-affinity, saturable, Na(+)-dependent transport system and is retained by the fibers primarily in that form. Our results suggest that the glia are not only the target of the glutamate signal released from non-synaptic regions of the crayfish medial giant axon during high-frequency stimulation, but that they are also the primary site of its inactivation. PMID- 10828543 TI - Looking back, looking forward: the legacy of the National Advisory Board on Ethics in Reproduction (NABER). AB - The National Advisory Board on Ethics in Reproduction (NABER) was an independent, private, not-for-profit board created in 1991 by the American College of Obstetricians and Gynecologists (ACOG) and the American Fertility Society (AFS) to review and comment on ethical issues related to the use of new reproductive technologies. This paper reviews NABER's mission and accomplishments, and attempts to make sense of why this vibrant, important organization closed its doors in 1998. PMID- 10828544 TI - The politics and ethics of human embryo and stem cell research. AB - There is great promise for stem cell research to develop cells and tissues for transplantation and treatment of diseases such as Alzheimer and Parkinson disease, diabetes, and heart problems. There is also promise to advance understanding and treatment of cancer and congenital defects. Human embryo research is fundamentally the only way to understand human fertilization, implantation, and early development. For years, federal funding of human embryo research has been held hostage to a congressional prolife agenda. Any reasonable solution to these political disputes that so greatly affect women's reproductive interests and the promise of health benefits from embryo and stem cell research should mandate that governmental sponsorship proceed. PMID- 10828545 TI - Ethics, politics, and human embryo stem cell research. AB - Three reports on ethical aspects of research involving human embryonic stem cells were issued in the final months of 1999. Two were from governmental agencies or commissions: the National Institutes of Health and the National Bioethics Advisory Commission. The third report was issued by the American Association for the Advancement of Science and the Institute for Civil Society. All three reports endorse the use of federal funds for embryonic stem cell research, but other differences distinguish these reports. This article describes the differences and provides an ethical analysis of the main arguments. PMID- 10828546 TI - Embryonic stem cells and the bigger reprogenetic picture. AB - In this country, public policy conversations about reproductive and genetic technologies have been conducted separately. This makes it difficult for policy makers and the public to notice the extent to which reproductive and genetic technologies are converging, and thus the extent to which we are approaching the possibility of reprogenetically shaping our children. Embryonic stem cell research offers a superb opportunity to identify that convergence, and thus to begin considering the creation of a public institution that would be devoted to contemplating the bigger picture of reprogenetically shaping children. PMID- 10828547 TI - Use of "excess" human embryos for stem cell research: protecting women's rights and health. AB - Proposed National Institutes of Health guidelines for stem cell research are too narrowly drawn and do not adequately protect the freedom of choice and health of women who donate embryos. They need to be expanded to cover not only the point of embryo donation, but also that of embryo creation. Guidelines are provided to ensure that donors undergoing hyperstimulation and egg retrieval gave voluntary informed consent to the production of embryos that might later prove in excess. A standard for determining when embryos have been overproduced is presented to address the possibility that additional embryos will be created for stem cell research in violation of the guidelines and at risk to women's health. PMID- 10828548 TI - What does "respect for embryos" mean in the context of stem cell research? AB - This article addresses two questions: 1) how to understand "respect for embryos," and 2) whether the distinction between surplus embryos from in vitro fertilization and embryos created for research purposes has validity. I caution against confusing respect for embryos as a form of human life with the respect that persons, as autonomous agents, deserve. I also argue that there is no moral difference between research that uses spare embryos and research that uses embryos created for that purpose. The value of the research is what determines whether it accords with the principle of respect for embryos, not the source of the embryos. PMID- 10828549 TI - Social ethics of embryo and stem cell research. AB - United States debates over stem cell research too often take the status of the embryo as the only decisive ethical issue, assume that if the embryo cannot be shown to be a person its destruction for a good cause is justified, and are insufficiently critical of economic incentives to legitimate and fund such research. Even if stem cell research ought not be banned, it could be situated and controlled on a spectrum of other health care needs through limited funding policies; aggressive peer review; regulatory and legal oversight applying to all research, however funded; and stringent patenting criteria. PMID- 10828550 TI - The ethical challenge of stem cell research. AB - This article analyzes the ethical issues raised by embryonic stem cell research and recent recommendations by the National Bioethics Advisory Commission (NBAC) regarding federal support for this research. The authors identify the key ethical issue as the moral significance that should be granted to early embryos and discuss arguments supporting the diverse answers to that question and the implications each view has on the formulation of rules and policies for stem cell research. The authors conclude that several of NBAC's recommendations regarding the derivation of stem cells from embryos for research are ethically justifiable and sound public policy. PMID- 10828551 TI - Our cells/ourselves: creating human embryos for stem cell research. AB - In September 1999, the National Bioethics Advisory Commission (NBAC) submitted its report, "Ethical issues in human stem cell research." The report recommends federal funding for stem cell research involving human embryos remaining after infertility treatment. It also suggests that at some later time, it may be appropriate for there to be federal funding for this same research using embryos expressly created for research purposes. This essay compares this recommendation to a similar recommendation reached 5 years earlier by the Human Embryo Research Panel of the National Institutes of Health. The NBAC recommendation is found to be much better packaged relative to the (contentious) goal of securing federal funding for this type of research. The merits of this goal are not discussed here. Instead, disappointment is expressed with the absence of any serious discussion of the ethical issues raised by the future possibility of stem cell research using research embryos. The report's silence on this question is significant given its promise to promote public debate on the profound ethical issues regarding human stem cell research. PMID- 10828552 TI - Ethics and politics of embryo and stem cell research: reinscribing the abortion debate. AB - Research on human embryos is controversial. Whether federal dollars can or should fund this work has been debated for 25 years, without satisfactory resolution. Many commentators point out that current conflict over embryo research grows out of this country's seemingly intractable conflict over abortion and the moral status of the fetus. This commentary reviews the most recent iteration of the embryo research debate--federal funding of human embryonic stem cell investigation--and explores the ways in which the terms of this debate not only reflect abortion disputes, but may in turn be shifted to strengthen feminist claims vis-a-vis abortion. PMID- 10828553 TI - Making babies in the 21st century: new strategies, old dilemmas. AB - Self-regulation of assisted reproduction by the medical community is not working. The continuing ban on in vitro fertilization (IVF) embryo research will only further erode the opportunity to develop comprehensive policies to protect the growing numbers of infertile couples, former cancer patients, gamete donors and their offspring produced through assisted reproduction. The President's National Bioethics Advisory Commission (NBAC) with the help of the Institute of Medicine can serve as an effective policy-building body because of its expertise, public forum, and national authority. National guidelines have been developed for gene therapy and xenotransplantation which could serve as models for dealing with the thorny bioethical dilemmas of assisted reproduction. In the interim, research on the social and ethical aspects of assisted reproduction should be a high priority for the NIH. PMID- 10828554 TI - Early separation and sibling incest. A test of the revised Westermarck theory. AB - This was a follow-up study of earlier reported findings by the present investigators suggesting, albeit equivocally, that separation during early childhood inhibited later sexual acts of a potentially procreative nature between siblings but did not deter other sexual activity. The present study surveyed 170 subjects, mostly in the Toronto area, by telephone and mail. Respondents reporting potentially procreative, postchildhood sexual acts (attempted or completed genital intercourse) with siblings were compared with those reporting sexual relationships excluding procreative acts, and a third sample reporting no postchildhood sibling sexual behavior. Consonant with expectations from the earlier study, prolonged separation during early childhood was associated with procreative postchildhood sexual activity but not with other postchildhood sexual activity. Contrary to predictions, however, both sexual activity groups reported significantly more nudity and physical contact with siblings during childhood than subjects reporting no sexual activity. The findings are discussed in terms of a revised version of the Westermarck hypothesis, which is consistent with a domain-specific approach to evolved incest avoidance mechanisms. PMID- 10828555 TI - The 2nd:4th digit ratio, sexual dimorphism, population differences, and reproductive success. evidence for sexually antagonistic genes? AB - The ratio between the length of the 2nd and 4th digit (2D:4D) is sexually dimorphic, with mean male 2D:4D lower than mean female 2D:4D. It recently was suggested that 2D:4D is negatively correlated with prenatal testosterone and positively correlated with prenatal estrogen. It is argued that high prenatal testosterone and low estrogen (indicated by low 2D:4D) favors the male fetus and low prenatal testosterone and high estrogen (indicated by high 2D:4D) favors the female fetus. The patterns of expression of 2D:4D are interpreted in terms of sexually antagonistic genes.We report data on the following. (a) reproductive success and 2D:4D from England, Germany, Spain, Hungary (ethnic Hungarians and Gypsy subjects), Poland, and Jamaica (women only). Significant negative associations were found between 2D:4D in men and reproductive success in the English and Spanish samples and significant positive relationships between 2D:4D in women and reproductive success in the English, German, and Hungarian samples. The English sample also showed that married women had higher 2D:4D ratios than unmarried women, suggesting male choice for a correlate of high ratio in women, and that a female 2D:4D ratio greater than male 2D:4D predicted high reproductive success within couples. Comparison of 2D:4D ratios of 62 father:child pairs gave a significant positive relationship. This suggested that genes inherited from the father had some influence on the formation of the 2D:4D ratio. Waist:hip ratio in a sample of English and Jamaican women was negatively related to 2D:4D. (b) Sex and population differences in mean 2D:4D in samples from England, Germany, Spain, Hungary (including ethnic Hungarians and Gypsy subjects), Poland, Jamaica, Finland, and South Africa (a Zulu sample). Significant sex and population differences in mean 2D:4D were apparent. PMID- 10828556 TI - Teaching may be hazardous to your marriage. AB - Kenrick et al.'s experiments demonstrate that men who view photographs of physically attractive women or Playboy centerfolds subsequently find their current mates less physically attractive and become less satisfied with their current relationships. What then would be the cumulative effect of being exposed to young, attractive women on a daily basis? Would there be any real consequences to the men's dissatisfaction with their relationships? Secondary school teachers and college professors come in contact with more young women at the peak of their reproductive value than others do. The analysis of a large, representative data set from the United States indicates that, while men in general are less likely to be divorced than women, and secondary school teachers and college professors in general are less likely to be divorced than others, simultaneously being male and being a secondary school teacher or college professor statistically increases the likelihood of being divorced (p <.05). We contend that the contrast effect that Kenrick et al. find in their experiments is cumulative and has real consequences. PMID- 10828557 TI - Is symmetry a visual cue to attractiveness in the human female body? AB - Small deviations from bilateral symmetry (a phenomenon called fluctuating asymmetry [FA]) are believed to arise due to an organism's inability to implement a developmental program when challenged by developmental stress. FA thus provides an index of an organism's exposure to adverse environmental effects and its ability to resist these effects. If one wishes to choose an individual with good health and fertility, FA could be used as an index of a potential partner's suitability. To explore whether this theory can be applied to human female bodies (excluding heads), we used a specially developed software package to create images with perfect symmetry. We then compared the relative attractiveness of the normal (asymmetric image) with the symmetric image. When male and female observers rated the images for attractiveness on a scale of 1 to 10, there was no significant difference in attractiveness between the symmetric and asymmetric images. However, in a two-alternative forced-choice experiment, the symmetric image was significantly more popular. The evidence suggests a role for symmetry in the perception of the attractiveness of the human female body. PMID- 10828558 TI - Evolved mechanisms underlying wayfinding. further studies on the hunter-gatherer theory of spatial sex differences. AB - Based on Silverman and Eals' hunter-gatherer theory of the origin of sex-specific spatial attributes, the present research sought to identify the evolved mechanisms involved in hunting that contribute to the dimorphism. The focus of these studies was the relationship between three-dimensional mental rotations, the spatial test showing the largest and most reliable sex difference favoring males, and wayfinding in the woods. Space constancy was presumed to be the evolved mechanism fundamental to both of these abilities. Measures of wayfinding were derived by leading subjects individually on a circuitous route through a wooded area, during which they were stopped at prescribed places and required to set an arrow pointing in the direction the walk began. As well, subjects were eventually required to lead the experimenters back to the starting point by the most direct route. In support of the hypotheses, males excelled on the various measures of wayfinding, and wayfinding was significantly related across sexes to mental rotations scores but not to nonrotational spatial abilities or general intelligence. PMID- 10828559 TI - Advances in vascular brachytherapy continue(1). PMID- 10828560 TI - Intracoronary radiation for patients with refractory in-stent restenosis: an analysis from the WRIST-Crossover Trial. Washington Radiation for In-stent Restenosis Trial. AB - BACKGROUND: In-stent restenosis (ISR) is a limitation of intracoronary stent implantation. In animal models and clinical trials, ionizing radiation has been shown to decrease in-stent neointimal formation. METHODS AND MATERIALS: In the Washington Radiation for In-stent Restenosis Trial (WRIST), patients with in stent restenosis were first treated with conventional catheter-based techniques and then randomized to gamma irradiation (l92Ir, 15 Gy at 2 mm from the source) or placebo. Patients randomized to placebo that developed recurrent ISR and presented with anginal symptoms were crossed-over to active therapy. We compared the clinical and angiographic outcomes of 39 patients in the crossover group with 65 patients treated primarily with radiation. The primary clinical endpoints were mortality, morbidity, and repeat target lesion revascularization (TLR) at 6 months. RESULTS: Procedural success was 100%. In-hospital and 30-day complications were minimal. At 6 months, the rate for any multiple adverse cardiac events (MACE: death, MI, or TLR) was 25.6% in the crossover group vs. 29.2% in the primary treatment group (p = 0.86). Three patients in the crossover group and four in the primary treatment group died (p = NS). There were no Q-wave myocardial infarction. Ten (9.6%) of the patients presented with late thrombosis and total occlusion: 6.2% (4/65) in the primary treatment group vs. 15.4% (6/39) in the crossover group (p = 0.13). CONCLUSIONS: Patients who fail conventional catheter-based intervention without radiation can be treated with 192Ir with results similar to those who initially receive brachytherapy. This is of specific importance with respect to patients initially assigned to the placebo arm of randomized clinical trials. PMID- 10828561 TI - Application of intravascular brachytherapy: utilization of beta-irradiation source delivery systems within a community-based cardiac center. AB - PURPOSE: There are numerous clinical studies ongoing to assess the outcome, physics, and radiobiology of intravascular brachytherapy and its effect on the reduction of the rate of restenosis after balloon angioplasty procedures. The present study reports on the experience of two different delivery systems as utilized in the community hospital setting. METHODS AND MATERIALS: Patients were enrolled into one of four ongoing trials at our institution: the Novoste Beta Cath trial, the Novoste Stents and Radiation Therapy trial (START), the Novoste START 40/20 trial, and the Guidant Intimal Hyperplasia Inhibition with Beta In stent Trial (INHIBIT). The Novoste studies utilized the Novoste Beta-Cath System with 90Sr/Y, and the Guidant INHIBIT trial used 32P. Enrollments into the various trials were determined by inclusion and exclusion criteria specified by each protocol. Randomization was conducted per criteria as determined by the participating study protocol. RESULTS: Forty-two patients were enrolled in total. Thirty-four were included in the Novoste trials and eight in the Guidant study, according to availability of the trial. Assessment of practicality of treatment was dependent primarily on treatment duration and extension of time of catheterization procedures by the addition of intravascular radiation. Average dwell time within the Novoste trials was 3 min 40 s, and 7 min 46 s for patients in the Guidant study. No acute complications were observed in any of the trials. CONCLUSIONS: Intravascular brachytherapy can be performed in the community hospital setting without compromising the efficiency of balloon angioplasty procedures. Pending long-term outcome data and FDA approval for specific delivery systems, endovascular brachytherapy in community hospital cardiac catheterization laboratories can be realized in an efficient and timely manner. PMID- 10828562 TI - Dose and dose rate effects of beta-particle emitting radioactive stents in a porcine model of restenosis. AB - BACKGROUND: Radioactive stents have been proposed as a means to prevent in-stent restenosis by inhibiting intimal proliferation with continuous low-dose irradiation. OBJECTIVES: The purpose of this study is to determine the effects of cumulative dose and dose-rate delivery on neointimal formation using 32P and 90Y beta-particle emitting radioactive stents in a porcine coronary model of restenosis. METHODS AND MATERIALS: We compared the late histologic results of 0.25 to 32.0 microCi 90Y (half-life 64 hours) (n = 64 stents) and 0.1 to 57.6 microCi 32P (half-life 14.3 days) (n = 55 stents) Beta-particle emitting radioactive stents with non-radioactive (n = 40) stents in a porcine coronary model of restenosis. A computer-based dosimetry modeling program was used to determine the 28 day cumulative dose and dose-rate delivery for the beta-particle emitting radioactive stents at a distance of 0.1 mm from the stent surface. RESULTS: Continuous low dose-rate (1 to 5 cGy/hr) radiation delivery for > 2 weeks via a 0.1 to 0.5 microCi 32P radioactive stent effectively reduced in-stent neointimal hyperplasia at 90 days. Cumulative doses of > 55 Gy induced severe adventitial fibrosis, microvascular damage and promoted the formation of a matrix rich neointima. Delayed vascular repair was evident at focal regions within the body of radioactive stents that delivered cumulative doses of > or = 140 Gy at 28 days and cumulative doses of 1,100 Gy at 90 days. CONCLUSIONS: These data may be useful in predicting safe and effective dose and dose rate delivery for beta particle emitting radioactive stents. PMID- 10828563 TI - Modulation of protein expression and activity by radiation: relevance to intracoronary radiation for the prevention of restenosis. AB - Restenosis is a common complication of percutaneous transluminal coronary angioplasty. Recent studies have demonstrated a striking reduction in the neointimal hyperplasia characteristic of restenosis following intracoronary radiation (IR), but the mechanisms by which radiation reduces neointima formation following balloon overstretch injury are not elucidated fully. In addition to direct antimitotic effects mediated via oxygen free radicals, ionizing radiation can induce the expression of numerous genes and thereby mediate indirect effects. Additionally, IR prevents restenosis at the cost of decreased healing and increased thrombosis, and we suggest that these adverse reactions can be modulated by adjunct pharmacology or gene-based strategies. This review discusses several genes and proteins modulated by radiation in the context of arterial injury, and their possible therapeutic relevance. PMID- 10828564 TI - External beam irradiation following balloon angioplasty in an atherosclerotic rabbit model. AB - PURPOSE: To study the effect of external beam irradiation on the morphometry of both angioplasted and nonangioplasted arteries in a hypercholesterolemic rabbit model. METHODS AND MATERIALS: Four groups of rabbit femoral arteries were studied: arteries (a) with no intervention, (b) irradiated with a 12-Gy x-ray dose, (c) treated with balloon angioplasty, and (d) dosed with 12 Gy 30 min after balloon angioplasty. RESULTS: External irradiation did not change vessel morphometry in nonangioplasted arteries. On the contrary, it induced neointimal formation and decreased luminal area, without causing any vessel remodeling in arteries treated with balloon angioplasty. CONCLUSION: External irradiation at 12 Gy given 30 min after angioplasty in the studied model accentuated the neointimal response to vascular injury, without causing any vessel remodeling. PMID- 10828565 TI - Dosimetry calculation for a novel phosphorus-32-impregnated balloon angioplasty catheter for intravascular brachytherapy. AB - PURPOSE: A phosphorus-32-impregnated balloon angioplasty catheter was used in a novel technique of simultaneous angioplasty and vessel irradiation. The 32P radionuclides were distributed on the surface of the balloon so that a certain amount of radiation was delivered while angioplasty was performed. Three dimensional dosimetry and dose-time relationship needs to be established for the catheter so that quantitative dosimetric information is available for both clinical treatment and research investigation. METHODS AND MATERIALS: The 32P impregnated balloon of an angioplasty catheter was assumed to have a cylindrical shape, and the radionuclides were assumed to be distributed uniformly on the curved surface of the cylinder. The dose rate at a point in space was computed by integrating the point dose-rate kernel of 32P over the radioactive surface of the balloon. The point dose-rate kernel was computed with Monte Carlo simulation of radiation transport. The energy spectra of 32P based on a mathematical model was used in the calculations. The three-dimensional dose distributions and dose-time relationships were calculated for balloons of various lengths and radii. RESULTS: At a short radial distance (e.g., 0.2 mm) away from the balloon surface, the dose distribution was uniform across a large portion of the balloon along the longitudinal axis, and dropped off rapidly at both ends of the balloon. Uniformity became worse as the radial distance increased. Uniformity was almost independent of balloon radius. The underdosed length at each end of the balloon was also almost independent of balloon length. In the central transverse plane, the dose reached a maximum at the surface of the balloon and then dropped off rapidly as the distance increases. Relative dose coverage outside the balloon was approximately independent of balloon radius and length, and the absolute dose coverage was approximately inversely proportional to balloon radius and length, assuming same total activity. CONCLUSIONS: Point dose-rate kernel of 32P beta emitter and the three-dimensional dose distributions of a 32P-impregnated balloon from an novel angioplasty catheter were calculated. A rule of thumb for dose calculation and dose coverage was established for simultaneous angioplasty and vascular brachytherapy with a 32P-impregnated balloon catheter. PMID- 10828566 TI - Measurement of density and calcium in human atherosclerotic plaque and implications for arterial brachytherapy. AB - PURPOSE: To measure density of arterial plaque specimens for purposes of improving calculation of intravascular radiation dose. METHODS AND MATERIALS: In the described technique, the mass of the specimen submerged in water is compared with its mass in air. Thirty-three plaque specimens harvested from cadavers and subsequently histologically classified (18 coronary, 15 noncoronary) were subjected to density measurement, and were also assayed for calcium using inductively coupled plasma optical emission spectroscopy (ICPOES). A dose point kernel (DPK) computer model extended to heterogeneous media is used to determine delivered dose to tissues for stents labeled with 32P, 103Pd, and 131Cs, based on measured density values. RESULTS: Plaque specimens identified histologically as noncalcified (non-class VII) had an average density of 1.22 +/- 0.03 g/cm3 (n = 19). Plaque specimens identified as calcified (Class VII) had an average density of 1.45 +/- 0.06 g/cm3 (n = 13). Density of calcified portions of plaque may be even higher because plaque specimens are heterogeneous. Plaque density was found to be correlated with calcium weight percentage (R2 = 0.67) and histologic percent area calcification (R2 = 0.58). Significant variations in calculated dose were found according to isotope, plaque density, and plaque thickness. The assumption of an "all water density" dose model overestimates dose to tissues. For 1-mm thick calcified (class VII) plaque, computed dose to tissues (via DPK model) are decreased by 29%, 34%, and 15%, for 32P, 103Pd, and 131Cs stents, respectively, compared with an "all water density" assumption model, when density is taken into account. Similar decreases are expected for catheter-based brachytherapy systems using beta or low energy (< 100 keV) gamma sources. CONCLUSIONS: This work has importance for radioactive stents and catheter-based brachytherapy due to dependence of dose on density at distances between 0.1 mm and 3 mm away from the radiation source. This dependence is important for both beta- and gamma-based systems. PMID- 10828567 TI - Intravascular irradiation using Re-186 liquid-filled balloon catheters: correlation between experimental and theoretical studies. AB - PURPOSE: Optimization of intravascular radiation to reduce stenosis following coronary angioplasty requires the ability to predict the patterns of radiation dose distribution. This investigation evaluated the agreement between Monte Carlo simulations and experimental radiation dose measurements for a radioisotope liquid-filled balloon catheter in a tissue equivalent phantom. METHODS AND MATERIALS: Direct measurements of the radiation dose from Re-186 liquid-filled balloons were made using thermoluminescent dosimeters (TLDs) and radiochromic film. Monte Carlo simulations were carried out using the Monte Carlo N-Particle code system (MCNP4B). RESULTS: The Monte Carlo generated dose values agreed with the experimentally determined results within the statistical uncertainty. A slightly higher penetration was indicated by regression analysis for the TLD data relative to the MCNP4B prediction that may be due to experimental configuration anomalies. For this balloon catheter, approximately 55 mCi of Re-186 will deliver 15 Gy at a 0.5 mm depth in tissue equivalent material in 5 min. CONCLUSIONS: Correlation between experimentally measured dose values and Monte Carlo computation supports the position that MCNP4B simulations constitute a valuable tool for investigating various clinical therapy designs. The agreement between Monte Carlo calculations and experiments provide confidence in applying MCNP4B to more sophisticated geometries of interest, and other methods of intravascular radiation dose delivery. PMID- 10828568 TI - Radioactive 32P-stent for in-stent restenosis in the porcine model. PMID- 10828569 TI - Reporting vascular brachytherapy: proposal for a revision of the AAPM TG 60 report. American Association of Physicists in Medicine. PMID- 10828570 TI - Physiological parameters evaluation following apnea in healthy premature infants. AB - To assess responses to central and obstructive apnea, we performed 10-hour polygraphic recordings in healthy 33-34 wGA infants. Each apnea period was paired with a control period. The presence of body movements (BM) and augmented breaths (AB), the EEG, heart rate (HR), respiratory rate, phase relationships between thoracic and abdominal respiratory movements, and changes in SaO(2) were evaluated. No apnea caused awakening. Apnea were not usually followed by motor events (no significant differences with control periods), with the exception of most obstructive apnea longer than 10 s. The mean percentage of changes following apnea, normalized to baseline values, was significant for the EEG (frequency slightly increased, mainly after obstructive apnea), HR (deceleration), and respiratory rate (increased). However, the mean values masked heterogeneity across apnea in the direction of the change in each parameter. The only significant correlation was between changes in HR and SaO(2). The increases in EEG frequency and respiratory rate seen in our study can be considered markers of CNS activation, but were small and inconsistent. The heart rate decelerations and SaO(2) changes are not activation markers. Thus, mechanisms underlying restarting of breathing efforts following apnea remain unclear in premature babies. Our investigation establishes the importance of using control data to distinguish between spontaneous and apnea-related events. PMID- 10828571 TI - Is a standard protocol necessary for oscillometric blood pressure measurement in term newborns? AB - OBJECTIVE: The necessity of taking only one randomized blood pressure measurement or averaging three repeated measurements and, the effects of various stages of the restful state and body position on blood pressure measurements obtained with the oscillometric technique were investigated in 138 healthy term newborns. METHODS: The Athena oscillometer was used to measure blood pressure. Three successive measurements with a 5-min interval were made in each of two positions, prone and supine, in random order 30 min after the last feeding if newborns were in very quiet or quiet sleep. During routine recording of vital signs, another (single) measurement was obtained before feeding the infant regardless of the body position of the newborn, provided that they were not struggling, crying or moving. RESULTS: For all systolic, diastolic and mean blood pressures measured, there were no significant differences among either prone, supine and single measurements or among three successive measurements in each position. CONCLUSIONS: We conclude that, in the routine care of term newborns, blood pressure measurements with the oscillometric technique may be made without the need of a special position or sleep state, provided that the measurements are made with an appropriate sized cuff in the absence of struggling, crying and movement of the newborn. Taking only one randomized measurement under these conditions would be enough and practical in daily newborn care practice instead of repeating and averaging many measurements. PMID- 10828572 TI - Bioactive transforming growth factor-beta in the lungs of extremely low birthweight neonates predicts the need for home oxygen supplementation. AB - Transforming growth factor-beta (TGF-beta) is a peptide implicated in tissue injury and repair but its role in the premature human lung remains unclear. In the present study, we used a TGF-beta responsive-promoter-luciferase construct in mink lung epithelial cells to quantify levels of biologically active TGF-beta (BA TGF-beta) in the endotracheal aspirate (ETA) fluid from 16 extremely low birthweight neonates [6 M/10 F, mean GA 26 weeks (range 23-30), mean BW 774 g (range 555-1,075)]. ETA fluid was obtained on day 1 and then every 4 days up to 32 days. BA-TGF-beta levels were low (92 +/- 19 pg/ml) in the first 24 h of life and then increased 5- to 10-fold with peak BA-TGF-beta levels (400 +/- 50 pg/ml) on day 20-25. BA-TGF-beta levels were higher in male than female infants (p = 0.0056). Prenatal steroids decreased significantly the amount of BA-TGF-beta recovered. High initial levels of BA-TGF-beta persisted over time and were predictive of the need for oxygen therapy at home. We conclude that abundant BA- TGF-beta is present in the lungs of preterm infants and speculate that it may be involved in inflammatory and repair processes encountered in acute and chronic lung disease. PMID- 10828573 TI - Spectral analysis of heart variability in the newborn infant. AB - We investigated the relationship between spectral power and both mean heart rate (HR) and heart rate variability (HRV). Spectral power was calculated using digital heart rate recordings from term infants. Regression analysis revealed a positive correlation between low-frequency (LF) sympathetic power and HR, and a negative correlation between high-frequency (HF) parasympathetic power and HR. HRV correlated positively in all regions of the power spectrum. In awake infants, the contribution of HF power to total power (HF/TP) was significantly decreased. LF power tended to be greater, however, this trend was not statistically significant. By following expected autonomic patterns, the findings of this study confirm that spectral analysis provides a noninvasive method for the assessment of autonomic activity influencing the newborn heart. The correlation between spectral power and HRV can serve as an additional tool in the study of autonomic dysfunction. PMID- 10828574 TI - Neonatal blood lymphocyte subpopulations: a different perspective when using absolute counts. AB - We compared the absolute counts of lymphocyte subpopulations in 15 neonates, and 9 adults using the whole lysed blood technique with 15 different triple immunostainings. To obtain accurate absolute lymphocyte counts in neonatal cord blood samples, the flow cytometric 'lympho-gate' was corrected for the erythroid cell contamination by normoblasts and unlysed erythrocytes. In contrast to earlier studies where relative frequencies were reported, we found that the major difference between neonatal and adult lymphocyte subpopulations concerned the much larger pool of naive 'untriggered' cells in neonates, standby for participation in primary immune responses. PMID- 10828575 TI - The efficacy of pentoxifylline as an anti-inflammatory agent in experimental Escherichia coli meningitis in the newborn piglet. AB - This study was done to evaluate the anti-inflammatory effect and the ensuing neuroprotective effect of pentoxifylline in neonatal experimental bacterial meningitis. Newborn piglets were divided into three groups: 10 in the control group (CG), 13 in the meningitis group (MG), and 13 in the meningitis with pentoxifylline group (PG). Meningitis was induced by intracisternal injection of 10(8) colony-forming units of Escherichia coli in 100 microl of saline. In PG, 20 mg/kg of pentoxifylline was given as a bolus intravenous injection 30 min before induction of meningitis and 6 mg/kg/h was given continuously throughout the experiment. In PG, the increase of CSF TNF-alpha level observed in MG was abolished. Reduced brain glucose and ATP concentrations observed in MG were significantly increased in PG. However, other parameters of inflammatory responses such as increased intracranial pressure, reduced glucose and increased lactate concentrations in the CSF observed in MG were not significantly down modulated. The extent of CSF leukocytosis was even higher in PG than in MG. Increased cerebral cortical cell membrane lipid peroxidation products and decreased Na(+),K(+)-ATPase activity observed in MG, indicative of meningitis induced brain cell membrane dysfunction, tended to improve without statistical significance in PG. In summary, although some anti-inflammatory effects have been observed, the overall anti-inflammatory effects of pentoxifylline was very weak, and it failed to significantly reduce the brain damage in experimental neonatal bacterial meningitis. PMID- 10828576 TI - Surfactant treatment effects on lung structure and type II cells of preterm ventilated lambs. AB - We evaluated surfactant treatment effects on lung morphology and alveolar type II cells of preterm ventilated lambs. Lambs were ventilated for 10 h following treatment of the right lung with natural surfactant. Lung parenchyma from the surfactant-treated right and the untreated left lung was compared morphometrically. Mechanical ventilation without surfactant resulted in distention of alveolar ducts accompanied by shallowing and loss of well-defined alveoli without disruption of collagen or elastin fibers. Surfactant treatment almost completely prevented these changes. The percent of normal parenchyma was 82 +/- 7% in surfactant-treated lobes and 26 +/- 5% in the nontreated lobes (p < 0.05). Type II cells became flatter in lungs ventilated without surfactant, and cell shape was preserved by surfactant treatment. The volume densities of lamellar bodies and multivesicular bodies in alveolar type II cells were not changed by surfactant treatment. With or without surfactant treatment, mechanical ventilation was associated with a shift in lamellar body distribution to a smaller size and a decrease in glycogen content of type II cells. Surfactant treatment of the preterm lung prevents alveolar distortion and atelectasis, but does not result in changes in subcellular organelles in immature type II cells. PMID- 10828577 TI - Contractile and relaxing mechanisms in pulmonary resistance arteries of the preterm fetal lamb. AB - Isolated pulmonary resistance arteries from term fetal lambs have nitric oxide (NO)- and prostaglandin-mediated relaxing mechanisms which are activated when PO(2) is raised from fetal to neonatal levels. The same vessels contract under hypoxia, and the contraction has been ascribed to endothelin-1 (ET-1). We have now studied these vasoeffector mechanisms before term (0.7 and 0.65 gestation) with the objective of determining whether their activity correlates with the development of susceptibility to oxygen changes. Experiments were carried out at neonatal PO(2), when expectedly relaxing mechanisms are maximally expressed, or under hypoxia. At either fetal age, the NO synthesis inhibitor, N(G)-nitro-L arginine methyl ester (100 microM), had no effect on basal tone, while indomethacin (2.8 microM) was a weak constrictor. Premature arteries did not contract when first exposed to hypoxia, but they responded marginally to a second exposure. The same arteries contracted strongly to a thromboxane A(2) analogue (ONO-11113, 0.1 microM) and ET-1 (10 nM), while their contraction to activating solution (5 mM Ca(2+) in K(+)-Krebs solution) was small and variable. At 0.7 gestation, bradykinin (0.1-100 nM), acetylcholine (0.01-10 microM), and sodium nitroprusside (0.1 nM to 10 microM) dose-dependently relaxed arteries precontracted with ONO-11113. Conversely, at 0.65 gestation the relaxation to bradykinin and acetylcholine was not dose-dependent and tended to be weaker. We conclude that preterm pulmonary arteries have viable effector mechanisms for contraction and relaxation. However, the capability for these mechanisms to be activated by PO(2) changes is markedly curtailed. PMID- 10828578 TI - Glutathione ethyl ester supplementation prevents mortality in newborn rats exposed to hyperoxia. AB - Human premature neonates suffer from respiratory distress syndrome due to immature lungs and require assisted ventilation with high concentrations of oxygen. Hyperoxic exposure and/or antioxidant deficiency causes an increase in the lung levels of reactive oxygen species (ROS) leading to oxidative stress induced cellular damage. In this study, we explored the protective role of the nonenzymatic antioxidant glutathione, by administering glutathione ethyl ester (GSHEE), in newborn rats exposed to hyperoxia (>95% FiO(2)). Our results show that GSHEE supplementation (5 mmol/kg/day) prevents mortality in newborn rats exposed to hyperoxia. We further show that delayed GSHEE supplementation in newborn rats, pre-exposed to hyperoxia for 4 days, also prevents death. Electron microscopic studies on the lung of GSHEE-treated hyperoxic rats showed normal histology and an absence of the marked swelling and degeneration of mitochondria and lamellar bodies, which are typically observed in the hyperoxic lungs of newborn rats. Furthermore, there were no apparent differences in weight gain or general appearance/activity among room air and hyperoxic GSHEE-supplemented animals when monitored, post-treatment, in room air for 30 days. Our results show a preventive/therapeutic role of GSHEE supplementation against mortality caused in newborn rats due to hyperoxic exposure, and may further be applicable to a variety of degenerative diseases that are caused as a result of ROS accumulation. PMID- 10828579 TI - Treatment of acute (Adult) respiratory distress syndrome. The holy grail of surfactant therapy. AB - The treatment of neonatal respiratory distress syndrome with surfactant represents a successful culmination of decades of basic and clinical research. In many babies, respiratory distress syndrome is a relatively pure expression of surfactant deficiency. Acute respiratory distress syndrome (ARDS) is a more common disease that is most frequently seen in adults, but the processes are common to lung injuries in newborns and children as well. While some impairment of production and secretion of surfactant constituents may be present in ARDS, surfactant inactivation is probably a more important factor in this disease. Until recently, surfactants available for human use have been easily susceptible to inactivation and this may explain why they have been less successful for treatment of ARDS than for neonatal respiratory distress syndrome. This review outlines recent information on surfactant inactivation and describes initiatives that may result in 'inactivation-proof' surfactants that may be of increased benefit in ARDS. PMID- 10828580 TI - Lung surfactant proteins A and D in innate immune defense. AB - The lung surfactant proteins (SP) A and D are large multimeric proteins and belong to a family of collagenous C-type lectins designated collectins. Both SP-A and SP-D are believed to play a role in the innate immunity of the lung. SP-A and SP-D bind to a broad spectrum of pathogens, including bacteria, viruses, fungi and yeasts but also lipopolysaccharides and allergens. Furthermore, SP-A and SP-D enhance the clearing of various pathogens by neutrophils and macrophages in vitro. Recent in vivo studies on SP-A deficient mice also support a role of SP-A in host defense. PMID- 10828581 TI - High-frequency oscillatory ventilation in the management of respiratory distress syndrome. AB - Early use of high-frequency oscillatory ventilation with a high volume strategy (HFOV-HVS) has been proposed to decrease the incidence of chronic lung disease following respiratory distress syndrome (RDS) in extremely immature infants. Despite encouraging results of animal experiments, clinical trials have provided discordant results. Our own multicenter trial showed that using HFOV-HVS, compared with conventional ventilation, decreased exogenous surfactant requirements, but did not modify pulmonary outcome, and increased the risk of severe intraventricular hemorrhage. This prompted us to change our management of RDS, by switching from elective use of HFOV to an 'early rescue' approach. PMID- 10828582 TI - Investigation of lung disease in preterm infants using magnetic resonance imaging. AB - Many preterm infants require ongoing respiratory support despite treatment with exogenous surfactant. The reasons for this are unclear, but may involve one or a combination of changes in water content or distribution within the lung. Detailed three-dimensional information with the potential to provide quantitation of water content may help to investigate this further. We aimed to determine if magnetic resonance (MR) imaging could be developed to study lung disease in preterm infants. Appropriate MR sequences and procedures were defined and we found that T1 and proton density weighted images could be successfully acquired. The images contained three-dimensional information that could not be obtained using chest radiograph. MR imaging may be a useful method for studying the pathology of respiratory distress syndrome and chronic lung disease in preterm infants. PMID- 10828583 TI - Report of the Fourth International Chromosome 6 Workshop 1999. 10-12 June 1999. Cambridge, UK. Abstracts. PMID- 10828584 TI - Assignment of type I phosphatidylinositol-4-phosphate 5-kinase (PIP5K1A) to human chromosome bands 1q22--> q24 by in situ hybridization. PMID- 10828585 TI - Assignment of the human postmeiotic segregation increased (S. cerevisiae) 1 (PMS1) to chromosome 2q31.1 by radiation hybrid mapping. PMID- 10828586 TI - Assignment of BUB3 to human chromosome band 10q26 by in situ hybridization. PMID- 10828587 TI - Assignment of GUCY1A3, a candidate gene for hypertension, to human chromosome bands 4q31.1-->q31.2 by in situ hybridization. PMID- 10828588 TI - Assignment of the ADAM15 gene to human chromosome band 1q21.3 by in situ hybridization. PMID- 10828589 TI - Assignment of the murine calcium-activated chloride channel genes Clca1 and Clca3 (alias gob-5) to chromosome 3 band H2-H3 using somatic cell hybrids. PMID- 10828590 TI - Assignment of the human metabotropic glutamate receptor gene GRM4 to chromosome 6 band p21.3 by radiation hybrid mapping. PMID- 10828591 TI - cDNA cloning and genomic structure of a novel gene (C11orf9) localized to chromosome 11q12-->q13.1 which encodes a highly conserved, potential membrane associated protein. AB - We have cloned and characterized a novel gene (C11orf9) mapping to chromosome 11q12-->q13.1. The transcript was initially identified as a partial cDNA sequence in the course of constructing a transcript map of the region between markers D11S1765 and uteroglobin known to encompass the gene causing Best disease. Using a combination of EST mapping, computational exon prediction, RT-PCR, and 5'-RACE its 5. 7-kb full-length cDNA sequence was subsequently obtained. The C11orf9 gene consists of 26 exons spanning 33.1 kb of genomic DNA and is located about 4.3 kb centromeric to FEN1. Biocomputational analysis predicts that its conceptual translation product of 1,111 amino acids contains two transmembrane helices as well as two proline-rich regions. Alignment reveals significant homology to hypothetical peptides from several other species including C. elegans and D. melanogaster, indicating a high degree of conservation throughout evolution. Northern Blot and RT-PCR analyses demonstrate widespread expression of a single transcript but varying degrees of abundance among the individual tissues tested. Mutation analysis of the entire coding sequence excluded C11orf9 as the Best disease gene. PMID- 10828592 TI - Assignment of claudin-1 (CLDN1) to human chromosome 3q28-->q29 with somatic cell hybrids. PMID- 10828593 TI - Assignment of the mouse Rbpsuh gene to chromosome 5 and one processed pseudogene Rbpsuh-rs3 to chromosome 6. PMID- 10828594 TI - Isolation and chromosomal assignment of a novel human gene, CORO1C, homologous to coronin-like actin-binding proteins. AB - We have isolated a gene, termed CORO1C (human coronin-like actin-binding protein 1C), that encodes a new member of the coronin-like family of proteins. The cDNA consists of 3,857 nucleotides, with an open reading frame of 1,422 bp encoding a 474 amino acid protein. The deduced amino acid sequence shared 65% identity with p57 (human coronin-like actin-binding protein), as well as 46% identity with coronin, a protein first isolated from the slime mold Dictyostelium discoideum. Computer analysis predicted that the product of the CORO1C gene would contain five WD repeats in its N-terminal region and a coiled-coil motif in its C terminal region, both of which are conserved among coronin-like proteins. CORO1C was ubiquitously expressed in all human tissues examined, in contrast to other known coronin-like molecules, each of which is expressed in a tissue-specific manner. Immunocytochemical staining demonstrated that CORO1C was co-localized with F-actin; therefore, the gene product is likely to be important in cytokinesis, motility, and signal transduction, as are the other members of this molecular family. We assigned this novel gene to chromosome 12q24.1 by fluorescence in situ hybridization. PMID- 10828595 TI - Expression map of human chromosome region 17p13.3, spanning the RP13 dominant retinitis pigmentosa locus, the Miller-Dieker lissencephaly syndrome (MDLS) region, and a putative tumour suppressor locus. AB - Chromosome region 17p13.3 is rich in genes, with 223 expressed sequence tags (ESTs) within the last 15 cM (7 Mb) of chromosome 17p in the GeneMap database. Loci for dominant retinitis pigmentosa (RP13), central areolar choroidal dystrophy (CACD), anterior polar cataract (CTAA2), Miller-Dieker lissencephaly syndrome (MDLS), and a region of tumour loss of heterozygosity (LOH) distinct from TP53 all map into the region adjacent to the 17p telomere. To date, however, there is no physical map of the region, which has resisted the efforts of the CEPH and Whitehead physical mapping programmes to generate contiguous clones across it. We have created a physical map covering approximately 3.5 Mb (6 cM)in this region, spanning the RP13 interval and extending distally to the gene MDCR (formerly, LIS1), which, when deleted, leads to the MDLS phenotype. The region covered is also the point of maximum LOH in lung cancer and has been implicated in the pathogenesis of many other human cancers. The map orders 47 sequence tagged sites, including 32 genes or ESTs, nine genetic markers, four anonymous sequences, and two YAC end clones, and highlights new candidate ESTs for involvement in RP13, MDLS, CTAA2, and a tumour-susceptibility gene. PMID- 10828596 TI - Cytogenetics and mechanisms of spontaneous abortions: increased apoptosis and decreased cell proliferation in chromosomally abnormal villi. AB - Genetic defects of the zygote, such as chromosome aberrations, are the most frequent causes of abnormal embryonic development and spontaneous abortion. However, the underlying mechanisms remain unknown. Chromosome aberrations likely cause changes in placental morphology and function (such as size, shape, vascularity, and the presence of trophoblastic inclusion). We postulated that chromosome aberrations may affect rates of cell proliferation or programmed cell death (apoptosis) during the differentiation of chorionic villi. To address these questions, we evaluated cell proliferation using a monoclonal antibody to Ki-67 (a cell-cycle marker) and apoptosis using the in situ end-labeling method (TUNEL) on paraffin-embedded placental tissues. Tissues were obtained from spontaneous abortions in early gestational periods with normal (11 cases) and abnormal karyotypes (15 cases), as well as eight normal control placentas from elective abortions. Apoptotic cells were found in the stroma of all cases, but were significantly higher in number in the stroma of chromosomally abnormal versus chromosomally normal spontaneous abortions. The apoptotic index of the trophoblasts was not significantly different between groups. Cell proliferation was higher in muscularized blood vessels in chromosomally normal placentas (both elective and spontaneous abortions) versus chromosomally abnormal spontaneous abortions. Cell proliferation was different in the trophoblast and stroma between the groups but to a lesser degree than in blood vessels. The morphological and biological data presented here suggest that: (1) chromosomally abnormal spontaneous abortions may occur because of different mechanisms than chromosomally normal spontaneous abortions, (2) apoptosis of the stromal cells and cell proliferation in blood vessels and stroma play an important role in the differentiation and functioning of villi, and (3) these changes could explain the etiology of spontaneous abortion and growth retardation of chromosomally abnormal embryos. PMID- 10828597 TI - Genomic organization, proposed alternative splicing mechanisms, and RNA editing structure of GRIK1. AB - In the present study, we report the genomic reconstruction of the glutamate receptor GluR5 gene (GRIK1, alias GLUR5; 402 kb) by the use of the data available in public databases. This analysis allowed characterization of the exon-intron boundaries, identification of the gene promoter region, presentation of the putative complete sequences of two GluR5 mRNA isoforms, and definition of the structure of GluR5 pre-mRNA surrounding the Q/R editing site. PMID- 10828598 TI - Identification and mapping of swine cyclin-dependent kinase inhibitor CDKN2A and CDKN2B exon 2 sequences. AB - We have isolated the swine homologs of human CDKN2A and CDKN2B exon 2 sequences. As in the human and mouse genomes, the exon 2 sequences of these two genes present a high level of sequence homology and are tightly linked. Using fluorescence in situ hybridization, we have mapped swine CDKN2A and CDKN2B to chromosome 1q25. This confirms the comparative mapping data among man, mouse, and swine, showing a conserved synteny among chromosome segments 9p21, 4C3-C6, and 1q25, respectively. PMID- 10828599 TI - Assignment of keratocan gene (KERA) to human chromosome band 12q22 by in situ hybridization. PMID- 10828600 TI - The eosinophil peroxidase gene forms a cluster with the genes for myeloperoxidase and lactoperoxidase on human chromosome 17. AB - Eosinophil peroxidase (EPX) is one of a family of mammalian peroxidases that includes myeloperoxidase (MPO), lactoperoxidase (LPO), and thyroid peroxidase (TPO). Here we show that the human EPX gene maps to chromosome 17q23.1, which localizes 34 kb from the LPO and MPO genes. Our results demonstrate that the EPX, LPO, and MPO genes form a cluster on human chromosome 17. PMID- 10828601 TI - Chromosome mapping of the human genes encoding the MAP kinase kinase MEK1 (MAP2K1) to 15q21 and MEK2 (MAP2K2) to 7q32. AB - Activation of the ERK mitogen-activated protein (MAP) kinase pathway has been implicated in the regulation of cell growth, differentiation and senescence. In this pathway, the MAP kinases ERK1/ERK2 are phosphorylated and activated by the dual-specificity kinases MEK1 and MEK2, which in turn are activated by serine phosphorylation by a number of MAP kinase kinase kinases. We report here the chromosomal localization of the human genes encoding the MAP kinase kinase isoforms MEK1 and MEK2. Using a combination of fluorescence in situ hybridization, somatic cell hybrid analysis, DNA sequencing and yeast artificial chromosome (YAC) clone analysis, we have mapped the MEK1 gene (MAP2K1) to chromosome 15q21. We also present evidence for the presence of a MEK1 pseudogene on chromosome 8p21. The MEK2 gene (MAP2K2) was mapped to chromosome 7q32 by fluorescence in situ hybridization and YAC clone analysis. PMID- 10828602 TI - Assignment of E-cadherin (CDH1) and KSP-cadherin (CDH16) to chromosome 16q22.1 by radiation hybrid mapping. PMID- 10828603 TI - Five human genes encoding F-box proteins: chromosome mapping and analysis in human tumors. AB - Members of the F-box protein (Fbp) family are characterized by an approximately 40 amino acid F-box motif. SCF complexes (formed by Skp1, cullin, and one of many Fbps) act as protein-ubiquitin ligases that control the G(1)/S transition of the eukaryotic cell cycle. The substrate specificity of SCF complexes is determined by the presence of different Fbp subunits that recruit specific substrates for ubiquitination. Unchecked degradation of cellular regulatory proteins has been observed in certain tumors and it is possible that deregulated ubiquitin ligases play a role in the altered degradation of cell cycle regulators. We have recently identified a family of human Fbps. As a first step aimed at determining if FBP genes could be involved in human neoplasia, we have mapped the chromosome positions of 5 FBP genes by fluorescence in situ hybridization (FISH) to 10q24 (BTRC alias beta-TRCP/FBW1a), 9q34 (FBXW2 alias FBW2), 13q22 (FBXL3A alias FBL3a), 5p12 (FBXO4 alias FBX4) and 6q25-->q26 (FBXO5 alias FBX5). Since most of these are chromosomal loci frequently altered in tumors, we have screened 42 human tumor cell lines and 48 human tumor samples by Southern hybridization and FISH. While no gross alterations of the genes encoding beta-Trcp/Fbw1a, Fbw2, Fbx4 and Fbx5 were found, heterozygous deletion of the FBXL3A gene was found in four of 13 small cell carcinoma cell lines. This is the first evaluation of genes encoding Fbps in human tumors. PMID- 10828604 TI - Identification and characterization of mouse orthologs of the AMMECR1 and FACL4 genes deleted in AMME syndrome: orthology of Xq22.3 and MmuXF1-F3. AB - The contiguous gene deletion syndrome AMME is characterized by Alport syndrome, midface hypoplasia, mental retardation and elliptocytosis and is caused by a deletion in Xq22.3, comprising several genes including COL4A5, FACL4 and AMMECR1. We have now cloned the murine Facl4 and Ammecr1 genes and have mapped both novel murine genes to mouse chromosome X band F1-F3. The murine and human orthologs show 96.5% (FACL4) and 95.2% (AMMECR1) identity at the amino acid level, with conservation of the respective putative subcellular localization signals. Our results show that Facl4 and Ammecr1 are the true murine orthologs of the human genes. Furthermore, the mapping of Facl4 and Ammecr1 to MmuXF1-F3 suggests that this subinterval is orthologous, at least for a portion of Xq22. 3. PMID- 10828605 TI - Assignment of the TCF-4 gene (TCF7L2) to human chromosome band 10q25.3. PMID- 10828606 TI - Chromosomal localization of the genes encoding ALDH, BMP-2, R-FABP, IFN-gamma, RXR-gamma, and VIM in chicken by fluorescence in situ hybridization. AB - Six structural genes encoding ALDH, BMP-2, R-FABP, IFN-gamma, RXR-gamma and VIM were mapped in the chicken by fluorescence in situ hybridization (FISH) using genomic and cDNA clones as probes. The genes were found to be located on four different macrochromosomes: chromosome 1 (IFNG and FABP), chromosome 2 (VIM and ALDH), chromosome 3 (BMP2) and a smaller macrochromosome, most probably chromosome 7 (RXRG). With the exception of IFNG none of the newly mapped sites corresponds to known orthologous regions between chicken and human chromosomes. PMID- 10828607 TI - Assignment of TTC4 to human chromosome band 1p31.3 and a pseudogene TTC4P to 7p14 ->p13 by in situ hybridization. PMID- 10828608 TI - The human multimerin gene MMRN maps to chromosome 4q22. PMID- 10828609 TI - Cloning, genomic sequence, and chromosome mapping of Scyb11, the murine homologue of SCYB11 (alias betaR1/H174/SCYB9B/I-TAC/IP-9/CXCL11). AB - A T-cell attracting CXC chemokine phylogenetically related to MIG and SCYB10 was recently characterized and termed SCYB11 (alias betaR1/H174/SCYB9B/I-TAC/IP 9/CXCL11). Here, we cloned the cDNA of the murine homologue of this protein, Scyb11, from interferon-gamma/lipopolysaccharide-stimulated RAW264.7 mouse macrophage-like cells. The nucleotide sequence of Scyb11 shares 63% identity with its human counterpart. It encodes a 100 amino acid immature protein of 11,265 Da which contains a putative signal peptide of 21 amino acids. The molecular mass of the mature protein was calculated to be 9,113 Da. Sequence identity of the murine and human SCYB11 proteins is 68%. Phylogenetic tree analysis of mouse CXC chemokines places SCYB11 together with the murine homologues of MIG and SCYB10 (Crg-2/muIP-10) on an individual branch. A genomic sequence was obtained by genome walking and subcloning DNA fragments from a BAC clone containing Scyb11. Like human SCYB11, Scyb11 contains 4 exons with intron/exon boundaries at positions comparable to the human gene. Whereas introns 2 and 3 are of similar length in the murine and human genes, intron 1 of Scyb11 contains 1,260 bp more than intron 1 of the human gene. Intron 1 of Scyb11 is also characterized by a 201-bp stretch with repetitive sequences of high cryptic simplicity. Using a BAC clone containing Scyb11, this gene could be mapped to chromosome 5 at position 5E3. Since human SCYB11 is localized on 4q21.2, this result confirms the mouse/human homology of the two chromosome regions. PMID- 10828610 TI - Assignment of developmentally regulated GTP-binding protein (DRG2) to human chromosome band 17p11.2 with somatic cell hybrids and localization to the Smith Magenis syndrome critical interval. PMID- 10828611 TI - Assignment of the plakophilin-2 gene (PKP2) and a plakophilin-2 pseudogene (PKP2P1) to human chromosome bands 12p11 and 12p13, respectively, by in situ hybridization. PMID- 10828612 TI - Assignment of the human metabotropic glutamate receptor gene GRM7 to chromosome 3p26.1-->p25.2 by radiation hybrid mapping. PMID- 10828613 TI - Delineation and physical separation of novel translocation breakpoints on chromosome 1p in two genetically closely associated childhood tumors. AB - Sporadic childhood tumors associated with Beckwith-Wiedemann syndrome (BWS) all show abnormalities of the same region on chromosome 11. In addition to chromosome 11, other chromosome regions are affected in some of these tumor types. In this study we analyzed the region on chromosome 1p involved in the etiology of BWS associated tumors, Wilms tumor, rhabdomyosarcoma, and hepatoblastoma. For this purpose we determined the location of two novel translocation breakpoints in this chromosome region in cells from a Wilms tumor and cells from a rhabdomyosarcoma. We constructed a map of the region and found that both breakpoints are separated by at least 875 kb. We identified a PAC clone which crosses the rhabdomyosarcoma breakpoint and found several exons within this clone. We established that this breakpoint is located proximal to the PAX7 gene and, therefore, identified a new region involved in the etiology of rhabdomyosarcomas. PMID- 10828614 TI - Comparative cytogenetics of hamsters of the genus Calomyscus. AB - Karyotypes of Calomyscus from different regions of Turkmenistan, Iran, and Azerbaijan were studied using chromosome banding (G- and C-banding) and analyses of meiosis in laboratory hybrids. Extensive variation in the diploid number and the number of autosomal arms (FNa) was revealed (2n = 30, FNa = 44; 2n = 32, FNa = 42; 2n = 44, FNa = 46; 2n = 44, FNa = 58; 2n = 37, FNa = 44; 2n = 50, FNa = 50; 2n = 52, FNa = 56). Centric and tandem fusions and heterochromatin changes were identified as the major modes of karyotype evolution in this group. Natural hybrids between individuals with different karyotypes were recorded, and regular chromosome pairing in meiosis was observed in laboratory hybrids. Fluorescence in situ hybridization with a 353-bp BspRI complex tandem repeat indicated that chromosomal repatterning occurred recently within the genus. There is no unequivocal evidence suggesting the role of chromosomal change in the speciation of the populations of Calomyscus examined. PMID- 10828615 TI - Isolation and characterization of a novel gene encoding a putative seven-span transmembrane protein, TM7SF3. AB - As part of a project involving large-scale sequencing of clones randomly selected from a human cDNA library, we isolated a novel human gene, termed TM7SF3 (transmembrane 7 superfamily member 3). Its open reading frame encodes a 570 amino acid protein containing seven putative transmembrane domains. The transcript of this gene was expressed in all human tissues examined, but most abundantlyin kidney. Immunocytochemical analysis demonstrated subcellular localization of TM7SF3 protein at the plasma membrane. We determined the chromosome location of TM7SF3 as 12q11.2-->q12 by a combination of fluorescence in situ hybridization and radiation hybrid mapping. PMID- 10828616 TI - Mapping of Zyx coding for zyxin in the rat and its exclusion as a candidate gene for lymphopenia. PMID- 10828617 TI - Assignment of the GPR14 gene coding for the G-protein-coupled receptor 14 to human chromosome 17q25.3 by fluorescent in situ hybridization. PMID- 10828618 TI - Assignment of the gene GRM1 coding for metabotropic glutamate receptor 1 to human chromosome band 6q24 by in situ hybridization. PMID- 10828620 TI - Assignment of dlk (Dlk1) to mouse chromosome band 12E-F1 by in situ hybridization. PMID- 10828619 TI - Identification of a novel beta-tubulin subfamily with one member (TUBB4Q) located near the telomere of chromosome region 4q35. AB - The human beta-tubulin supergene family consists of several isotypes with many associated pseudogenes. Here we report the identification of yet another beta tubulin sequence designated TUBB4Q. This tubulin maps 80 kb proximal to the facioscapulohumeral muscular dystrophy (FSHD1) associated D4Z4 repeats on chromosome 4q35. The genomic structure contains four exons encoding a putative protein of 434 amino acids. The TUBB4Q nucleotide and protein sequence show 87% and 86% homology to beta2-tubulin, respectively. Although the genomic structure shows all functional aspects of a genuine gene, no transcript could be detected. TUBB4Q-related sequences were identified on multiple chromosomes. Since these sequences mutually exhibit a high nucleotide sequence homology, they presumably belong to a novel subfamily of beta-tubulin genes. Although the chromosome 4q35 tubulin-member probably represents a pseudogene, ectopic expression due to a postulated position effect variegation (PEV), makes TUBB4Q an ideal dominant negative candidate gene for FSHD1. PMID- 10828621 TI - Assignment of WDR7 (alias TRAG, TGF-beta resistance associated gene) to orthologous regions of human chromosome 18q21.1-->q22 and mouse chromosome 18D.1 E.3 by fluorescence in situ hybridization. PMID- 10828622 TI - Assignment of mimecan gene (OGN) to human chromosome band 9q22 by in situ hybridization. PMID- 10828623 TI - Assignment of Slc11a3 to mouse chromosome 1 band 1B and SLC11A3 to human chromosome 2q32 by in situ hybridization. PMID- 10828624 TI - Identification and expression analysis of C15orf3, a novel gene on chromosome 15q21.1-->q21.2. AB - We have isolated C15orf3, a novel human gene that lacks homology to any known gene family. The C15orf3 gene encodes a transcript of 1676 nt with an ORF of 187 amino acids and a predicted protein product size of 20.8 kDa. Northern blot analysis showed ubiquitous expression in adult tissues. EST database searching revealed the presence of C15orf3 homologs in rat and mouse. C15orf3 was mapped to chromosome 15q21.1-->q21.2 using the Stanford G3 radiation hybrid panel. PMID- 10828625 TI - Cytomegalovirus infection in normal and immunocompromised humans. A review. AB - Although cytomegalovirus (CMV) disease is a severe complication among immunocompromised patients, its cutaneous features have not been reported frequently. CMV belongs to the Herpesviridae family sharing with the other members the ability to remain latent in their natural hosts after an initial infection and to produce overt disease in several settings. The natural history of human CMV infection is characterized by primary infection, latent infection and reinfection. This article reviews the extremely variable aspects of the clinical presentation of CMV infection in normal and immunocompromised humans, focusing on the dermatological manifestations, and indicates the laboratory tests for detecting CMV responsibility in skin disorders. PMID- 10828626 TI - Elevated serum KL-6 levels in patients with systemic sclerosis: association with the severity of pulmonary fibrosis. AB - BACKGROUND: Serum KL-6 has been suggested to be a useful marker for the evaluation of interstitial lung disease activity. OBJECTIVE: To determine the correlation between serum KL-6 levels and pulmonary fibrosis in patients with systemic sclerosis (SSc). METHODS: Serum samples from patients with limited cutaneous SSc (lSSc; n = 19), diffuse cutaneous SSc (dSSc; n = 26) and normal individuals (n = 15) were examined by ELISA. RESULTS: Serum KL-6 levels in SSc patients were significantly higher than those in normal controls. KL-6 levels in dSSc patients were significantly elevated compared with those in lSSc patients. Elevated KL-6 levels were associated with the presence of pulmonary fibrosis in SSc patients or dSSc patients. Furthermore, KL-6 levels inversely correlated with percentages of diffusion capacity of carbon monoxide and vital capacity in SSc patients or dSSc patients. CONCLUSION: KL-6 may be a simple, serologic indicator for the severity of pulmonary fibrosis in SSc. PMID- 10828627 TI - Cutaneous monomorphous CD4- and CD56-positive large-cell lymphoma. AB - BACKGROUND: Recently, CD56 (NCAM)-positive lymphomas, such as nasal and nasal type angiocentric NK/T cell lymphoma, aggressive NK cell leukemia/lymphoma and blastic NK cell lymphoma, were described by several authors as a unique group of lymphoma. OBJECTIVE: In this study, we intend to clarify the clinicopathological features of cutaneous CD4+ and CD56+ lymphoma. METHODS: Four patients with cutaneous CD4+ and CD56+ lymphoma were studied. RESULTS: Age at the first examination ranged from 71 to 89 years (mean = 81.2 years). One patient was female and 3 were males. The organ mainly involved at presentation was the skin. Lymphadenopathy, splenomegaly, leukemic spread and central nervous system involvement were observed as the disease progressed. The mean survival time was 12.2 months. Epstein-Barr virus was not detected within the tumor cells. CONCLUSION: This peculiar lymphoma is different from nasal and nasal-type angiocentric NK/T cell lymphoma and aggressive NK cell leukemia/lymphoma. Similar cases have been reported as blastic NK cell lymphoma/leukemia. PMID- 10828628 TI - Renal amyloidosis in recessive dystrophic epidermolysis bullosa. AB - BACKGROUND: Although it is known that renal amyloidosis may complicate several dermatoses, recessive dystrophic epidermolysis bullosa (RDEB) complicated by nephropathy has been thought to be rare. We, however, had seen a young adult with RDEB who died of renal failure due to systemic amyloidosis. OBJECTIVE: A retrospective study was performed in order to investigate the incidence and etiology of renal amyloidosis in RDEB. METHODS: Routine urinalysis, serum amyloid A protein (SAA) and creatinine levels were repeatedly determined in 11 patients with RDEB (mean age 17.7 years, range 5-28, 7 males, 4 females). Nephropathy was defined as the presence of both proteinuria and hematuria with red blood cell casts. RESULTS: Seven out of 9 generalized RDEB patients had nephropathy including 3 cases with end-stage renal disease (2 died within 2 years from the onset of nephropathy), while 2 patients with localized RDEB did not. Levels of SAA were significantly higher in patients with nephropathy than those in patients without nephropathy (p<0.05). CONCLUSION: Nephropathy is a common and serious complication of RDEB. Renal amyloidosis may play an important role in its etiology. We recommend that patients with RDEB should be periodically screened for nephropathy due to amyloidosis by urinalysis and measuring SAA levels. PMID- 10828629 TI - An X-linked gene involved in androgenetic alopecia: a lesson to be learned from adrenoleukodystrophy. AB - BACKGROUND: Adrenoleukodystrophy (ALD), including its adult variant adrenomyeloneuropathy (AMN), is an X-linked recessive trait characterized by progressive demyelinization of the nervous system. The gene defect involves a peroxisomal transporter protein, resulting in accumulation of very-long-chain fatty acids in the brain and other organs such as the adrenal glands. Affected men show various endocrine disorders. Moreover, disturbances of hair growth are frequently mentioned in reports on ALD/AMN. OBJECTIVE: This study was performed to delineate further the hair status and type of hair loss in men with AMN. METHODS: We examined and documented the status of hair growth in 16 men suffering from AMN. A meticulous history with particular regard to hair changes was taken from all patients and their family members. RESULTS: The age of the patients varied between 27 and 62 years, their mean age was 39.8 years. Twelve men showed male-pattern androgenetic alopecia (AGA), Hamilton grades IV-VIII, 3 men had a female-pattern AGA (Ludwig grade I or II). Ten of the patients with male-pattern AGA had reached Hamilton stage VII or VIII. The remaining scalp hair was unusually scarce and thin in 11 cases, regardless of the grade of AGA. Moreover, in 10 of 16 patients the eyelids showed pronounced madarosis. The remaining body hair was found to be normal. If present, endocrine manifestations had started prior to the onset of alopecia, and in 11 of 12 patients hair loss was apparent before neurological symptoms were noted. CONCLUSION: ALD/AMN gives rise to two different types of hair loss. Firstly, affected men show diffuse hair loss involving the entire scalp and the eyelashes. Secondly, they tend to develop AGA more frequently and earlier and in a severer form. Paradoxically, pronounced AGA is present although the patients may simultaneously show some degree of hypogonadism. Hence, the X-linked ALD mutation can be taken as a well-defined gene within the polygenic spectrum of genes responsible for AGA. This may be of theoretical importance for the elucidation of the pathogenetic pathways of AGA. PMID- 10828630 TI - Male-pattern hair loss in Norwegian men: a community-based study. AB - BACKGROUND: A Norwegian community study was conducted as an add-on to a large general health survey in Trondlag County, Norway, to characterize the cross sectional relationship of patient perceptions with the degree of hair loss. METHODS: All members of the community were invited to participate in the general health survey and male participants, aged 20-50 years (n = 7,250), regardless of their degree of hair loss, were asked to complete a mail-based questionnaire. The questionnaire measured satisfaction with hair appearance, self-reported degree of hair loss, self-assessment of Norwood/Hamilton hair pattern, bother due to hair loss as well as concern about getting older due to hair loss, and included the SF 12 general health status questionnaire. RESULTS: Respondents (n = 4, 101) rated their hair loss on a 7-point, textual scale that ranged from 'a full head of hair' to 'I am bald'. The majority (63%) of participants reported at least a little hair loss and 27% reported moderate to severe loss. Using the Norwood/Hamilton hair patterns, participants rated themselves as class II (25.5%), III (8.6%), IV (8. 8%) or V or worse (19.5%). Relative to men without hair loss, a greater proportion of men with hair loss reported being bothered (20. 5 vs. 2.3%), concerned about growing older (43.4 vs. 10.6%) and dissatisfied with overall hair appearance (22.6 vs. 94%, all p< or =0.001). Men with hair loss also reported greater levels of perceived noticeability of hair thinning to others. CONCLUSIONS: A high proportion of Norwegian men aged 26-50 years self reported having at least some hair loss. Results suggest that men who perceive themselves as having greater hair loss are more bothered by their hair loss, more dissatisfied with the appearance of their hair and have greater concern about getting older and losing more hair. PMID- 10828631 TI - Minocycline-induced lupus. A systematic review. AB - BACKGROUND: Minocycline has increasingly been associated with different adverse auto-immune reactions including drug-induced lupus. OBJECTIVE: To identify the scope of minocycline-induced lupus and to characterise its typical features. METHODS: Comprehensive Medline and Embase search of the English and non-English literature for case reports of minocycline-induced lupus. RESULTS: We included 57 cases of minocycline-induced lupus (mean age +/- SD at onset: 21.6+/-8.6 years, median time of exposure: 19 months, range 3 days to 6 years). All patients showed the clinical features of polyarthralgia/polyarthritis often accompanied by liver abnormalities. Twelve patients had evidence of dermatological manifestations (i.e rash, livedo reticularis, oral ulceration, subcutaneous nodules, alopecia). The ANA test was positive in all patients. CONCLUSION: Long-term exposure to minocycline may be associated with drug-induced lupus. Baseline and periodic liver function and ANA tests accompanied by appropriate clinical monitoring are suggested for patients receiving long-term minocycline therapy. PMID- 10828632 TI - Continuous topical administration of a petrolatum formulation by a novel disposable diaper. 1. Effect on skin surface microtopography. AB - BACKGROUND: Cutaneous problems are commonly associated with the use of diapers. Aiming to help reduce them, we have explored the use of the inner layer of diapers as a means to deliver to the skin dermatological formulations intended to help protect it from overhydration and irritation. OBJECTIVE: To determine the feasibility of using the inner layer of the diaper as a vehicle for topical delivery of a petrolatum-based formulation and to determine its impact on skin surface microtopography. METHODS: Two independent, blinded, randomized clinical trials were conducted, on children 16-24 months of age. All comparisons were done versus a control diaper, identical to the test product except for the absence of the petrolatum formulation. The studies determined the effects of the novel diaper on transfer of formulation to the skin and skin surface microtopography. RESULTS: During normal diaper use, formulation transfer from the diaper to the skin occurred in a cumulative, time-dependent manner and use of the formulation treated diaper was associated with significant reductions in skin surface roughness compared to the control diaper. CONCLUSIONS: The results demonstrated the feasibility and skin surface benefits associated with continuous topical administration of a petrolatum-based formulation by this novel diaper. This unprecedented dosimetric approach offers new avenues to reduce further the dermatological problems commonly associated with diaper use. PMID- 10828633 TI - Continuous topical administration of a petrolatum formulation by a novel disposable diaper. 2. Effect on skin condition. AB - BACKGROUND: Diaper dermatitis is a common childhood affliction. Aiming to help reduce the prevalence of this problem, we have developed a novel diaper to deliver to the skin dermatological formulations intended to help protect the skin from overhydration and irritation. OBJECTIVE: To determine the clinical benefits of a novel disposable diaper designed to deliver a petrolatum-based formulation continuously to the skin during use. METHODS: Two independent, blinded, randomized clinical trials were conducted, involving an aggregate total of 391 children, 8-24 months of age. All comparisons were done versus a control diaper, identical to the test product except for the absence of the petrolatum formulation. The studies determined the effects of the novel diaper on skin erythema and diaper rash. RESULTS: Use of the formulation-treated diaper was associated with significant reductions in severity of erythema and diaper rash compared to the control product. CONCLUSIONS: The results demonstrated the clinical benefits associated with continuous topical administration of a petrolatum-based formulation by this novel diaper. We anticipate that this advance in diaper design will contribute significantly to further reduce the prevalence and severity of irritant contact dermatitis in the diaper area. PMID- 10828634 TI - Strontium nitrate decreased histamine-induced itch magnitude and duration in man. AB - BACKGROUND: Previous studies have demonstrated that strontium salts have a potent and broad ability to suppress sensory irritation. OBJECTIVE: To ascertain the possible antipruritic effects of topical strontium salts, we conducted a double blind randomized study to evaluate the effect of 20% strontium nitrate on itch magnitude (intensity) and duration. METHODS: In 8 human subjects, strontium nitrate was applied to the volar forearm and vehicle control to the contralateral volar forearm for 30 min; both substances were reapplied for an additional 5 min. Itch was then experimentally induced by intradermal injection of histamine. Itch magnitude was rated each minute for the first 20 min using a visual analogue scale (VAS). Itch duration in minutes was also recorded. RESULTS: Strontium nitrate, in comparison to its vehicle control, significantly shortened itch duration from 28.1+/-5.4 min (mean +/- SEM) to 18.5+/-4.2 min (p<0.01) and reduced itch magnitude at time points 12-20 min and overall (p<0.05). CONCLUSION: Strontium nitrate may act as a topical antipruritic agent in reducing histamine mediated itch. PMID- 10828635 TI - Male-pattern baldness in men with X-linked recessive ichthyosis. AB - BACKGROUND: X-linked recessive ichthyosis (XRI) is a genetic disorder of keratinization with extracutaneous manifestations due to deficiency of steroid sulfatase (STS). Because STS plays an important role in androgen metabolism, and elevated levels of dehydroepiandrosterone sulfate have been reported in young men with andogenetic alopecia (AGA), the hypothesis was advanced that men with XRI do not show AGA or develop only mild forms of common baldness. METHODS: Patients with a diagnosis of XRI confirmed by analysis of the microsomal sulfatases in our clinic between 1984 and 1998, and in whom study of the case histories depicted the typical clinical presentation of XRI, were sent a questionnaire with the Hamilton-Norwood scale of patterns of hair loss in men, inquiring them to designate the condition of their scalp hair. RESULTS AND CONCLUSIONS: Of 15 questionnaires returned, 7 indicated Hamilton-Norwood I, 3 Hamilton-Norwood II, 3 Hamilton-Norwood III-vertex, 1 Hamilton-Norwood IV and 1 Hamilton-Norwood VII. The results of this survey do not support the hypothesis that XRI and AGA are mutually exclusive, in as much as advanced AGA was found among these men. PMID- 10828636 TI - Primary mucinous carcinoma of the scalp. AB - Primary cutaneous mucinous carcinoma (MC) is a rare epithelial neoplasm derived from the sweat glands. Herein, we report a case of MC located on the head. A 66 year-old woman underwent excision of a nodular tumor with a reddish brown surface on the left parietal region. Histopathology revealed a neoplasm extending from the reticular dermis into the subcutaneous fat. The tumor cell aggregates showed cribriform and solid lobules and were embedded in lakes of mucin, separated by thin, fibrous septae. Focally single neoplastic cells were arranged in an Indian file pattern. The tumor cells displayed an eosinophilic cytoplasm, large basophilic nuclei and some discrete nuclear atypia. Vascular spaces, filled by densely packed erythrocytes between the septae, were also observed. We compared the mucinous component with the tumor cell and the stromal component by light microscopy. Analyzing the tumor by an image analysis system in Alcian-blue stained serial sections, we found the averaged total tumor area measuring 99.7 mm(2). The area of the mucinous component measured 92.4 mm(2), that of the tumor cells 3.7 mm(2) and that of the stromal component 3.6 mm(2). The extensive checkup of the patient disclosed no evidence for a further malignant neoplasm. After excision of the tumor an adjuvant radiotherapy was performed. The patient was free of recurrence and metastatic spread of the mucinous carcinoma during a 4 year follow-up. PMID- 10828637 TI - Segmental manifestation of Darier disease. What is the genetic background in type 1 and type 2 mosaic phenotypes? AB - Darier disease is an autosomal dominant disorder which may occasionally become manifest in a segmental form. Two clinical phenotypes with a different genetic background have been elaborated in recent years. More than 50 patients with isolated linear disease expression have been documented. In this phenotype the skin outside the segmental affection is absolutely normal. Such a phenotype is explained by a postzygotic mutation with somatic mosaicism which was labeled as type 1 manifestation of segmental forms in autosomal skin disorders. A patient with classical type 1 segmental Darier disease is presented. On the other hand, only 3 patients with Darier disease showing a segmental manifestation in combination with a diffuse distribution have so far been observed. These cases correspond to the recently described type 2 manifestation of segmental forms of autosomal dominant disorders. We describe a fourth patient with type 2 segmental Darier disease. The genetic explanation of such a phenotype is possible with the assumption that a germline mutation for the disease exists but, in addition, a postzygotic mutation is needed resulting in loss of heterozygosity. Hence, in a circumscribed region a homozygous or hemizygous state of the mutation is apparent which can explain the enhanced severity of the segmental manifestation. PMID- 10828638 TI - Keratoacanthoma developing on a pigmented patch in incontinentia pigmenti. AB - Cutaneous manifestations of incontinentia pigmenti (IP) have classically been described as three sequential stages: an initial vesicobullous stage, a verrucous stage and a stage of swirled pigmentation. Verrucous lesions tend to last longer than vesicobullous eruptions, often persisting until 1 year of age. However, adult patients with verrucous lesions are rare. We report a case of keratoacanthoma with marked dyskeratosis on a pigmented patch in a 20-year-old woman. This tumor, like subungual keratotic tumors of IP, might have been developed as one of the late manifestations of the disease. PMID- 10828639 TI - A case of symptomatic heterozygous female Fabry's disease without detectable mutation in the alpha-galactosidase gene. AB - We report a case of symptomatic heterozygous female Fabry's disease with low alpha-galactosidase blood activity. We could not find any mutations in the coding regions of either the signal peptide or the enzyme subunit in our case. PMID- 10828640 TI - Photo-accentuated eruption and vascular deposits of immunoglobulin A associated with hepatitis A virus infection. AB - Skin rash associated with hepatitis A virus infection has rarely been reported. We describe a patient with hepatitis A virus infection who presented a rubelliform rash markedly accentuated in sun-exposed areas; direct immunofluorescence studies of the lesion revealed immunoglobulin (Ig) A deposition on the endothelial cells in the upper dermis. Oral rechallenge tests of the previously administered drugs failed to reproduce the eruption. The preferential setting of immune complexes containing IgA at sites of sun exposure and sunlight as a triggering factor might have been responsible for the development of the eruption in this patient. Eruptions associated with hepatitis A virus infections may be more frequent than commonly thought. Because of difficulty to exclude the possibility of drug eruptions, these cases might have been overlooked. In patients with such a disorder, a careful clinical workup such as IgM antibodies for hepatitis A virus at diagnosis and during follow-up is especially recommended. PMID- 10828641 TI - A figurate papulosquamous variant of inflammatory vitiligo. AB - The classic description of inflammatory vitiligo is an erythematous rim at the periphery of a patch of hypopigmented or depigmented skin. The histological correlate is a superficial perivascular infiltrate of mononuclear cells. However, we observed a 61-year-old patient with inflammatory vitiligo who had thin solid pink scaly plaques as well as serpiginous lesions with fine scale. Histologically, a lichenoid infiltrate was seen as was a thickened stratum corneum with parakeratosis. In a review of the literature, scattered case reports of similar findings were identified, either in the Japanese and French literature or from over 25 years ago. The clinical and histological spectrum of inflammatory vitiligo should be expanded to include solid and annular papulosquamous plaques as well as lichenoid infiltrates with exocytosis. PMID- 10828642 TI - A case of viral warts with particular fibrillar intracytoplasmic inclusion bodies. AB - A new type of skin wart was observed in a Japanese patient. It was characterized by intracytoplasmic inclusions with a 'fibrillar' structure which were distinct from previously described wart-associated inclusions. The papillomavirus (HPV) group-specific antigen could be detected, but DNA hybridization and PCR amplification using probes or PCR primers specific for the main skin HPV genotypes (including HPV-63 which is also associated with 'filamentous' inclusions) were negative. We consider that this cytopathic effect could correspond to an HPV genotype which has not yet been characterized. PMID- 10828643 TI - Food-dependent exercise-induced anaphylaxis occurring only in winter. PMID- 10828644 TI - Lipodystrophia centrifugalis abdominalis infantilis with ulceration. PMID- 10828645 TI - Dapsone-responsive persistent erythema multiforme. PMID- 10828646 TI - Juxtaclavicular beaded lines: A malformative condition affecting sebaceous glands. PMID- 10828647 TI - Meta-analysis of clinical trials with copolymer 1 in multiple sclerosis. AB - Glatiramer acetate (copolymer 1) was licensed in the USA in 1996 for the treatment of relapsing-remitting multiple sclerosis. In order to assess its efficacy, a meta-analysis of all randomized controlled trials was performed. Two double-blind studies, accounting for a total number of 299 patients, contribute to this analysis. Patients undergoing the treatment have decreased probability of relapse at 12 months (OR 0.17, 95% CI 0.05-0.51, p = 0.002) and of unsustained progression of disability at 24 and 35 months (OR 0.57, 95% CI 0. 34-0.95, p = 0.031, and OR 0.50, 95% CI 0.28-0.90, p = 0.019). These data suggest that glatiramer acetate represents an alternative to interferon treatment in relapsing remitting multiple sclerosis. However, further evidence of efficacy is required to justify its use in clinical practice. PMID- 10828649 TI - Bleeding and subsequent anemia: a precipitant for cerebral infarction. AB - BACKGROUND AND OBJECTIVES: The relationship between bleeding and subsequent anemia (BSA) and the occurrence of stroke has not been sufficiently studied. The purpose of the present study was to elucidate the characteristics of stroke associated with BSA. METHODS: We studied 16 consecutive patients with acute stroke associated with anemia (hemoglobin level on admission < or =9.0 g/dl) and compared their stroke subtypes with those of 32 control subjects. RESULTS: The cause of anemia was upper gastrointestinal bleeding in 11 patients (ulcers in 8; carcinomas in 2, and hemorrhagic gastritis in 1), bleeding from a hemorrhoid in 2, uterine cervical bleeding in 1, ecchymosis probably related to medication in 1, and chronic blood drainage in 1. At least 10 patients had a history of recent (<1 week), active bleeding. Clinical and imaging studies showed that all the patients had infarcts and none had intracerebral hemorrhages. Thirteen patients had infarcts in the region of the middle cerebral artery (MCA) (total MCA region in 2; partial, cortical area in 5; subcortical area in 5, and lenticulostriate artery region in 1), 2 had anterior cerebral artery (ACA) region infarction, and 1 had cerebellar infarction. All 11 patients who underwent vascular imaging studies showed significant stenosis and/or occlusion of the internal carotid artery (ICA) (n = 5), the MCA (n = 4), both the ICA and MCA (n = 1), or the ACA (n = 1). Of the different stroke subtypes (large vessel infarction (LVI), small vessel infarction, cardiogenic embolic infarction, intracerebral hemorrhage), LVI was significantly (p<0.05) more frequent in patients with stroke associated with BSA than in the controls, even though the demographics and risk factors were similar in each group. CONCLUSIONS: The close temporal relationship between the bleeding and the onset of stroke, as well as the predominance of the LVI subtype in the BSA-associated group as compared to controls, suggest that BSA may precipitate atherothrombotic cerebral infarction. A hemodynamic alteration, enhanced thrombosis or a combination of these appears to be the pathogenic mechanism. PMID- 10828648 TI - New hypointense lesions on MRI in relapsing-remitting multiple sclerosis patients. AB - BACKGROUND: Preliminary observational studies with multiple sclerosis (MS) patients have reported strong correlations between an increase in hypointense lesion load (black holes) on T1-weighted spin echo images, and an increase in disability. OBJECTIVE: We assessed the relationship of hypointense lesions to the clinical course of disease among 50 relapsing-remitting MS patients in the controlled setting of a randomized clinical trial. METHODS: Fifty patients with relapsing-remitting disease were enrolled in a randomized double-blind two-arm (cladribine vs. placebo) clinical trial of 1-year duration. All patients had monthly clinical evaluations and MRIs over the course of the trial. Multivariate techniques were used to identify predictors of clinical severity from information on exacerbations, MRIs, baseline clinical parameters, and demographics. RESULTS: At baseline, clinical severity is weakly related to counts of black holes, with rank correlations between counts and clinical scores (EDSS and SNRS) of absolute magnitude 0.3. Rates of appearance of new black holes over the course of the trial are higher for patients with more severe disease at baseline (EDSS > or = 4) than for the less severe patients. Changes in clinical severity over the course of the trial are best predicted by baseline neurologic scores and numbers of exacerbations, with black holes adding no further improvement in prediction. CONCLUSIONS: Numbers of exacerbations seem more critical to short-term clinical outcomes in relapsing-remitting MS, as reflected by patients' clinical scores, rather than black holes. Various imaging methods and MRI indices capture complementary information relating to MS disease processes. The determination of which processes are affected by different drugs should lead to more effective treatment of MS patients. PMID- 10828650 TI - Primary intracerebral hemorrhages in the Besancon stroke registry. Initial clinical and CT findings, early course and 30-day outcome in 350 patients. AB - The purpose of this study was to estimate the risk factors, early course, outcome and neuroimaging patterns in primary intracerebral hemorrhages (PIH). Using the Besancon Stroke Registry, 350 patients with first PIH documented by computed tomography (CT) between 1987 and 1993 were included in the present study. Patients with hemorrhage secondary to traumatism, brain tumor, thrombolytic treatment, vascular malformation or with hemorrhagic infarction were excluded. All CTs were evaluated to define the location, extension and volume of bleeding (55% of CT were performed within the first 12 h). Causes of death were classified and the 30-day outcome survival was evaluated with a modified Rankin scale (40 patients underwent a noncodified surgical procedure and were excluded from the outcome evaluation). Locations were lobar (36.5%), lenticular (32%), thalamic (15.7%), cerebellar (8.8%), midbrain and pons (2%), intraventricular (2%), caudate (1%) and multiple (2%). Risk factors included hypertension (54.8%), alcohol (18%) anticoagulant treatment (8.8%) and none (31.2%). The largest mean volume was in putaminal (41.7 ml) and lobar (39.8 ml) locations. Among 191 patients admitted before the 12th hour of evolution, 51 (26.7%) experienced an early clinical worsening. In this group, the percentage of patients with anticoagulant treatment (19.6%) was significantly higher (p<0.0001). PIH enlargement was documented in 3 patients. Overall, the 30-day mortality rate was 24.2% with 48% of all deaths occurring in the first 3 days. Death and 30-day survival status were closely associated with PIH volume (p<0.0001). Our study provides information on the natural history of PIH and especially on initial evolution. PIH volume seems to be an interesting indicator for death and functional status at 30 days. PMID- 10828651 TI - Increased expression of laminin 1 in the skin of amyotrophic lateral sclerosis. AB - Abnormalities of collagen and glycosaminoglycans of skin have been reported in patients with amyotrophic lateral sclerosis (ALS). However, little is known concerning laminin in ALS. The aim of this study is to evaluate laminin 1 immunohistochemistry of skin in ALS patients. We studied laminin 1 immunoreactivity of skin from 17 patients with ALS, 17 patients with other neurological or muscular diseases (control group A) and 10 patients without neurological or muscular disorders (control group B). The dermis, the epithelial basement membrane of the epidermal layer and the basement membrane of skin appendages and blood vessels of skin in ALS patients were significantly higher than in control groups A and B, and showed a significant positive correlation with the duration of illness in the optical density for laminin 1 immunoreactivity. In addition, there was an appreciable positive relationship in the laminin 1 immunoreactivity between the dermis and the basement membrane in ALS patients. These data suggest that alterations of laminin 1 in the skin of ALS patients could be related to the pathogenesis of ALS and may contribute to changes in the mechanical properties of the skin, resulting in the absence of bedsores in ALS patients. PMID- 10828652 TI - Nerve conduction studies in multiple system atrophy. AB - To study the frequency and severity of peripheral neuropathy in multiple system atrophy (MSA), we performed nerve conduction studies in 42 MSA patients suffering from either cerebellar MSA (MSA-C) or parkinsonian MSA (MSA-P). Abnormal nerve conduction was present in 24% of the patients. Abnormalities were significantly more frequent in MSA-P (43%) compared to MSA-C (14%). Motor nerve conduction velocities were reduced in 4% of the MSA-C and in 7% of the MSA-P patients. Abnormal compound muscle action potentials were more frequent in MSA-P (29% versus 7% in MSA-C) pointing to a more pronounced loss of motor axons in this subgroup. Sensory nerve conduction velocities were abnormal in 4% of the MSA-C and 14% of the MSA-P patients, and mean sensory nerve action potentials were normal in all MSA-C and reduced in 7% of the MSA-P patients. The data provide evidence that the peripheral nervous system is differentially affected in MSA-C and MSA-P. PMID- 10828653 TI - Blood pressure elevations in riluzole-treated patients with amyotrophic lateral sclerosis. AB - OBJECTIVE: To determine whether riluzole is associated with blood pressure elevations in patients with amyotrophic lateral sclerosis (ALS). BACKGROUND: Though previously reported, hypertension is not considered a frequent adverse effect of riluzole. METHODS: We reviewed data from 35 consecutive ALS patients on riluzole, and 88 randomly selected controls without and 20 patients with ALS who were not on riluzole. RESULTS: A significantly greater number of ALS patients on riluzole had blood pressure elevations (28 of 35 patients) compared to controls (26 of 88, p<0.001; 8 of 20, p = 0. 007). Median systolic and diastolic blood pressures were both significantly higher in riluzole-treated (140/86 mm Hg) than in control patients without ALS (120/70 mm Hg, p<0.001). Systolic, but not diastolic, blood pressures were significantly higher in riluzole-treated patients than in controls with ALS (126 mm Hg, p = 0.002). CONCLUSIONS: Riluzole treatment may be associated with mild blood pressure elevations. Future prospective trials of riluzole should closely assess hypertension. PMID- 10828654 TI - Increased cerebrospinal fluid tau protein in multiple sclerosis. AB - Axonal damage is now being recognized as a common finding in multiple sclerosis (MS) lesions and a cause of irreversible neurological damage. Attempts to identify markers of early axonal damage are of great significance. This prompted us to examine the microtubule-associated protein tau in the cerebrospinal fluid (CSF) of patients with MS vs. controls. Tau was measured by double antibody sandwich ELISA. Increased CSF tau levels were found in MS as compared to controls (medians 249.6 and 135 pg/ml respectively, p<0.001). Half of the MS patients presented with levels above the upper limit of the controls. A significant increase vs. controls was found in both relapsing-remitting and progressive subtypes. These data may indicate axonal impairment in a subpopulation of MS patients and may provide a tool for the estimation of axonal damage during life. PMID- 10828655 TI - Developmental dyslexia: re-evaluation of the corpus callosum in male adults. AB - Using a new method based upon the measurement of four angles, we analyzed the corpus callosum of 23 adult male dyslexics and 25 age-matched controls on MRI sagittal scans. Two out of the four angles measured showed significant differences between the groups that are consistent with previous findings concerning the size of the corpus callosum in dyslexics. In particular, posterior regions are concerned, displaying a lowered corpus callosum in dyslexics. These findings are discussed in relation to cortical parietal asymmetries and possible hormonal events, referring to the testosterone hypothesis, that could occur in utero or in the perinatal period in the dyslexic population. PMID- 10828656 TI - Benign monomelic amyotrophy of the lower limb--case report and brief review of the literature. PMID- 10828657 TI - Simultaneous brachial diplegia and rotational vertigo due to combined spinal anterior and vertebrobasilar embolism. PMID- 10828658 TI - Erdheim-Chester disease with spinal cord manifestations. PMID- 10828659 TI - Spontaneous intraspinal epidural hematomas--report of 2 cases and review of the literature. PMID- 10828660 TI - Acute Wernicke's syndrome mimicking brainstem stroke. PMID- 10828661 TI - Risk factors for urinary incontinence in women. AB - OBJECTIVES: To review the epidemiological evidence on risk factors for urinary incontinence (UI) in women. METHODS: Using Medline and a manual search we identified the main papers published in English up to 1998. RESULTS: There is consistent evidence that the frequency of UI increases with age, but there is little information on the frequency of different types of UI in different age groups. With regard to other risk factors, there is a clinical consensus that vaginal delivery and, more in general, obstetric traumas are associated with UI, although epidemiological studies are not always consistent. Less defined is the role of race, menopausal status, history of gynecological surgery, body mass index, smoking, and coffee and alcohol consumption on the risk of UI. CONCLUSION: In the coming years, epidemiological research should focus particularly on the potential differences in the epidemiological characteristics of different types of UI in order to gain insight into the pathogenic mechanisms. PMID- 10828662 TI - p53 expression predicts progression and poor survival in T1 bladder tumours. AB - OBJECTIVES: Histological grade (G) is the only parameter proved to have prognostic value for progression in T1 transitional cell carcinoma (TCC) of the bladder, although it is considered inaccurate to make clinical decisions on individuals. The aim of the present study was to evaluate the prognostic relevance of p53 expression in T1 TCC of the bladder. METHODS: Clinical records of 207 patients with T1 TCC of the bladder were reviewed for clinical parameters reported to influence the evolution of superficial bladder cancer. Among these 207 patients, 40 developed muscle-invasive disease (20 G2 and 20 G3). A retrospective case-control study was then carried out comparing the latter 40 tumours with 40 control tumours matched by grade, sex, age, number and size of the tumours, chemical exposure and presence of carcinoma in situ. p53 immunostaining with monoclonal antibody was performed in these two groups. RESULTS: Histological grade was the only clinical parameter that influenced evolution. p53 expression correlated with tumour progression, since it was observed in 21 out of 24 p53-positive tumours and in only 20 of 56 p53-negative tumours (p<0.0001), showing a specificity of 93. 5% and a sensitivity of 53%. p53 expression correlated as well with patient survival, being 39% in patients with p53-positive tumours and 80% in patients with p53-negative tumours at 60 months (p<0. 0001). CONCLUSIONS: p53 protein expression has prognostic value for survival and progression in T1 bladder tumours and can be used for early detection of poor-prognosis T1 bladder tumours. PMID- 10828663 TI - Prognostic value of MIB-1 antibody labeling index to predict response to Bacillus Calmette-Guerin therapy in a high-risk selected population of patients with stage T1 grade G3 bladder cancer. AB - PURPOSE: Proliferation rate is an important determinant of bladder tumor progression. However, this factor has not yet been correlated to bacillus Calmette-Guerin (BCG) therapy response in a selected high-risk population of patients with stage T1 grade G3 bladder cancer. To assess the predictive value of the proliferation rate, an immunoreactivity test with monoclonal antibodies MIB-1 was carried out. The aim of this study was to evaluate the prognostic value of an MIB-1 labeling index by selecting a group of responsive patients prior to intravesical therapy. MATERIALS AND METHODS: After complete transurethral resection, 35 patients with T1G3 bladder carcinoma received 6 weekly installations of BCG (intravesical Pasteur strain: 75 mg in 50 ml course of BCG therapy). After treatment a cystoscopy and randomized biopsies of the bladder mucosa were carried out and all recurrences were systematically resected. All tissue samples were fixed in Bouin's solution, embedded in paraffin and stained with hematoxylin-eosin-safran. Pathologists had sufficient material to perform immunomarking in 25 patients using peroxidase-antiperoxidase (PAP) technique, with antiprotein monoclonal antibody MIB-1 (Immunotech, Marseilles, France) to study MIB-1 expression before BCG therapy. Consensus was obtained from three independent pathologists for all sections. The results were expressed in a percentage of marked nuclei. Ten percent increment thresholds were established from 10 to 60%. Contingency tables were established, chi2 (p1) and Fisher exact test (p2) were performed for each threshold of 10%. RESULTS: Median follow-up was 57.3 months (range 25-144). Of the 25 patients, 8 (32%) did not respond to BCG therapy, 17 (68%) responded positively. With a 20% threshold, there was a statistical difference (p1 = 0.03, p2 = 0.04) between responder (R) and nonresponder (NR) patients. All the 7 patients with less than 20% of nuclear activity positively responded to BCG. At this threshold level, sensitivity was high but specificity low (positive predictive value = 0.44). If we consider other reactivity thresholds there were no statistical differences between R and NR patients (10%) threshold p1 = 0.13, p2 = 0.19; 30% p1 and p2 = 0.20; 40% p1 = 0.82, p2 = 0.61; 50% p1 = 0.57, p2 = 0.55). CONCLUSION: Our study indicates that the proliferation rate, assessed by MIB-1 immunoreactivity in Bouin's solution fixed primary tissue, could be a useful predictive marker of outcome in T1G3 bladder carcinoma. With a 20% reactivity cut-off, a negative MIB-1 immunomarking appears to predict a positive response to BCG instillations. However, on the other hand, MIB-1 is of limited clinical use because the low specificity of this test cannot predict failure and then select candidates for cystectomies. PMID- 10828664 TI - Glutathione S-transferase null genotypes in transitional cell bladder cancer: a case-control study. AB - OBJECTIVES: This study was conducted (1) to examine whether the GSTM1 and GSTT1 null genotypes are risk factors for bladder cancer, and (2) to study a possible association of these genotypes with disease severity. METHODS: This case-control study was undertaken over a 21-month period and included 89 newly diagnosed transitional cell bladder cancer patients and 147 controls; both patients and controls originated from a defined population (residents of the loannina region, Northwestern Greece) and were similar with regard to mean age, male to female ratio and smoking habits. The GSTM1 and GSTT1 genotypes were identified by multiplex polymerase chain reaction on peripheral blood DNA samples. Genotype frequencies among patients and controls were assessed and the association of the genotypes with tumor grade and stage at presentation were statistically evaluated by the chi(2) test. RESULTS: The GSTM1 null genotype was strongly associated with bladder cancer. The odds ratio, attributable and population attributable risks were estimated at 2.76, 0.64 and 0.40, respectively. The correlation between the GSTM1 null genotype with stage, although not statistically significant, was estimated at an odds ratio of 2.6 for invasive disease. The correlation of GSTM1 null genotype with tumor grade did not yield a statistically significant result. The GSTT1 null genotype was not statistically associated with bladder cancer. CONCLUSION: According to our study, individuals with the GSTM1 null genotype carry a substantially higher risk for bladder carcinogenesis. The GSTM1 null genotype is not associated with more aggressive disease in terms of tumor grade, although there is a correlation between this genotype and stage of the disease. PMID- 10828665 TI - Absence of urethral post-void milking: an additional cause for incontinence after radical prostatectomy? AB - OBJECTIVE: Urinary incontinence following radical prostatectomy is thought to be mainly due to stress leak as a result of sphincter insufficiency or detrusor dysfunction. However, a number of patients complain of stress-independent urinary leakage following voiding, i. e. a post-micturition dribble, of uncertain origin. In order to establish wether post-micturition dribble is related to altered post void milking in the urethra, voiding cystourethrograms (VCUGs) were performed before and after radical prostatectomy and correlated with the presence of post micturition dribble. METHODS: 23 VCUGs were recorded before and 19 VCUGs at 10-15 days following radical prostatectomy. A standard questionnaire regarding urinary symptoms was given to all patients pre- and postoperatively at defined intervals. RESULTS: 12 of 19 patients (63%) had post-void urethral milking prior to surgery, none of these reported post-micturition dribble. 6 of the 7 patients (86%) without post-void urethral milking reported post-micturition dribble. Postoperatively only 1 of 16 patients (6%) had post-void urethral milking. Of the 15 patients without postoperative urethral milking, 13 (87%) reported post micturition dribble. The decrease in rate of milking and increase in rate of post micturition dribble from before to after surgery was statistically significant (chi(2) test, p = 0.0001 and p<0.0001, respectively. CONCLUSIONS: These data suggest that post-void milk-out of the urethra is often absent in the early postoperative period after radical prostatectomy and that this is associated with post-micturition dribble. Aside from detrusor and sphincter dysfunction, urethral dysfunction, i.e. the absence of urethral post-void milking, seems to be an additional cause of incontinence following radical prostatectomy. PMID- 10828666 TI - Discrepancy between Gleason scores of biopsy and radical prostatectomy specimens. AB - OBJECTIVE: The grade of the prostate cancer is an important factor in defining prognosis and deciding on treatment. In this study, we compared the Gleason score determined by 18-gauge core needle biopsies with both the Gleason score and pathological staging of the radical prostatectomy specimens. PATIENTS AND METHODS: Between July 1992 and September 1998, we performed 144 radical retropubic prostatectomies for clinically localized prostatic carcinoma, after a negative frozen section in bilateral pelvic lymphadenectomy in all cases. Ten patients with pathologic stage T1a and T1b were excluded. The final study group consisted of 134 patients, all of whom had been diagnosed with adenocarcinoma by transrectal needle biopsies with an 18-gauge automated spring-loaded biopsy gun. No patients received neoadjuvant therapy, including androgen deprivation and radiation therapy. All patients had a designated Gleason score on the needle biopsy and prostatectomy specimens. RESULTS: We found that grading error was greatest with well-differentiated (Gleason score 2-4) tumors, The accuracy was 15% for Gleason score 2-4 on needle biopsy. Of the 113 evaluable patients with Gleason score 5-7 on needle biopsy, 110 (97%) were graded correctly. All of the Gleason score 8-10 on needle biopsy was graded correctly. But only 1 patient in our series had Gleason score 8 on needle biopsy. Twenty-seven (25%) of 110 patients with a biopsy grade of Gleason score <7 had the cancer upgraded to 7. Of patients with both Gleason score <7 in the needle biopsy and Gleason score 7 in the prostatectomy specimen, only 3 (11%) had tumor confined to the prostate. CONCLUSION: The potential for grading error is greatest with well-differentiated tumors and of patients with both Gleason scores <7 in the needle biopsy and Gleason score 7 in the prostatectomy specimen, only 11% had tumor confined to the prostate. This effects treatment policy, especially for watchful waiting criteria. PMID- 10828667 TI - Transition zone biopsy in the detection of prostate cancer. AB - OBJECTIVE: About 25% of all prostate cancers occur in the transition zone (TZ). We analyzed the impact of 4 systematic TZ and 2 systematic apex (AP) biopsies in addition to systematic sextant biopsies in an effort to establish the diagnostic importance of early prostate cancer. METHODS: One hundred and thirty patients underwent systematic transperineal multipoint prostate biopsy (biopsy of 12 sites, including 4 TZ and 2 AP biopsies). RESULTS: Forty-one of 130 men (31.5%) had biopsy specimens positive for cancer, and cancer originated in the TZ alone in 4 of these 41 patients (9.8%). Fourteen patients underwent radical retropubic prostatectomy. We compare the pathological findings of radical prostatectomy specimens and biopsy results. Prostate cancers predicted to be stage T(2c) by TZ biopsy were all classified as pT(2c) or greater. CONCLUSIONS: Routine TZ biopsy does not substantially increase the prostate cancer detection rate; however, it can be useful in patients who require repeat biopsy. PMID- 10828668 TI - Safety, efficacy and impact on Patients' quality of life of a long-term treatment with the alpha(1)-blocker alfuzosin in symptomatic patients with BPH. The Italian Alfuzosin Co-Operative Group. AB - OBJECTIVE: The aim of this study was to assess the impact of a 12-month treatment period with alfuzosin, 2.5 mg t.i.d, on symptomatic patients with BPH (n = 355) by means of the International Prostate Symptom Score (I-PSS), the Symptom Problem Index (SPI), the BPH Impact Index (BII), and the single Quality of Life (QoL) question proposed by the WHO. STUDY DESIGN: This was a naturalistic study, carried out under conditions of normal practice by 22 centres of urology. METHODS: At baseline and on a quarterly basis up to 12 months of treatment, subjective and objective (uroflowmetry and residual urine volume) responses of the patients were evaluated. The appearance of adverse medical events (AMEs) was carefully monitored and recorded throughout the trial. RESULTS: Both the BII and the Qol question improved gradually over time (60 and 54. 6%, respectively, after 12 months of treatment). At any visits, the improvements were statistically significant versus the baseline (p<0. 01). Alfuzosin was able to improve the BPH symptoms progressively and significantly over time: total mean score I-PSS: 3rd month = 22. 7%, 6th month = 38.4%, 9th month 0.50%, 12th month = 61.6%. The improvement was more marked in patients with severe symptoms at baseline (I-PSS score 20-35, 63.6%). A progressive and marked improvement over time of the problems due to symptoms (SPI) was observed in the whole population (61.7% after 12 months of treatment). After 12 months of treatment, uroflowmetric data showed a significant increase in peak (+5.5 ml/s) flow rate, associated with a marked decrease in residual volume: -31 ml (-53.5%). Twenty-five patients (7.1%) experienced one or more AMEs (total AMEs n = 44). Globally, 14 vasodilatory events and 30 non-vasodilatory events were reported. Fifteen (4.3%) patients dropped out prematurely from the study for safety reasons. Seven serious AMEs were reported during the study period. CONCLUSIONS: This study showed that long term treatment with alfuzosin in usual clinical practice had a continued and positive impact on the patients' QoL. PMID- 10828669 TI - Long-term outcome of transrectal high- intensity focused ultrasound therapy for benign prostatic hyperplasia. AB - OBJECTIVE: The aim of this study was to determine the long-term outcome after transrectal high-intensity focused ultrasound (HIFU) therapy for patients with lower urinary tract symptoms (LUTS) due to benign prostatic hyperplasia (BPH). METHODS: Between June 1992 and March 1995, 98 men (mean age: 66 years) with LUTS due to BPH underwent transrectal HIFU therapy at our institution, and the data of 80 patients were included in this long-term analysis. Principal inclusion criteria were a peak flow rate (Qmax) < or =15 ml/s, AUA/IPSS score > or =18 and a prostate volume < or =75 ml. Postoperatively, patients were seen at 6-month intervals with assessment of symptom score, uroflowmetry and post-void residual volume. In the present analysis, follow-up was terminated at 4 years. The mean follow-up of the study population (excluding the patients who underwent transurethral resection of the prostate, TURP, due to insufficient therapeutic response) was 41.3 months (range: 13-48 months). RESULTS: In treatment responders (HIFU only; n = 45), the symptom score decreased from preoperatively 19.6 to 8.5 (-53%) after 12 months and subsequently showed only marginal fluctuations within the 4-year study period. The Qmax increased from preoperatively 9.1 to 11.8 ml/s (+30%) after 12 months and gradually declined to 10.2 ml/s (+12%) after 4 years. 35 men (43.8%) underwent TURP due to insufficient therapeutic response during the 4-year study period. The mean time interval between HIFU therapy and TURP was 26.5+/-2.7 months (range: 1-48 months). The retreatment-free period was significantly longer for patients with a pretreatment average flow rate >5 ml/s (p = 0.05) and lower grades of urodynamically documented bladder outflow obstruction (p = 0.03). A similar trend, which did not reach statistical significance, was noted for individuals with higher Qmax and lower post-void residuals. CONCLUSIONS: These long-time data indicate that transrectal HIFU therapy for BPH, at least in its present form, did not stand the test of time, as 43.8% of patients had to undergo TURP within 4 years after initial therapy. These data underline the need for long-term studies with follow-ups over several years to reliably assess the role of less invasive treatment options for BPH. PMID- 10828670 TI - Two-year results of transurethral resection of the prostate versus four 'less invasive' treatment options. AB - OBJECTIVE: The aim of this study was to compare the efficacy or transurethral resection of the prostate (TURP) versus four less invasive treatment options during a 2-year follow-up. MATERIAL AND METHODS: 95 elderly men with lower urinary tract symptoms due to benign prostatic hyperplasia (BPH) were assigned prospectively to the following five treatment arms; transurethral resection of the prostate (TURP; n = 28), transurethral electrovaporization (TUVP; n = 17), visual laser ablation of the prostate (VLAP; n = 17), transrectal high intensity focused ultrasound (HIFU; n = 20) and transurethral needle ablation (TUNA); n = 15). Preoperative workup included the International Prostate Symptom Score (IPSS), uroflowmetry, post-void residual volume (PVR), prostate volume determined by transrectal ultrasonography and a multichannel pressure flow study. Postoperative follow-up at 6, 12, 18 and 24 months included assessment of IPSS, PVR and uroflowmetry. RESULTS: At study entry, patients assigned to one of the five treatment arms were comparable with respect to age, peak flow rate (Q(max)), IPSS, prostate size and the degree of bladder outflow obstruction. During study, 1 patient in the TURP group (4%) required a secondary TURP, as compared to 23.5% (n = 4) after TUVP, 26.7% (n = 4) after VLAP, 15% (n = 4) after HIFU and 20% (n = 3) following TUNA. In patients not subjected to a secondary procedure, the IPSS decreased a mean 13. 9 after TURP, as compared to 12.7 after TUVP, 12.9 after VLAP, 7.0 after HIFU, and 9.8 after TUNA. Q(max) increased 11.5 ml/s (mean) after TURP, as compared to 11.1 ml/s after TUVP, 5.6 ml/s after VLAP, 2.5 ml/s after HIFU and 2.3 ml/s after TUNA. CONCLUSION: In up to a quarter of the patients, a secondary TURP is performed within the first 2 years after 'less invasive' procedures. These data underline the need for long-term studies to reliably assess the role of less invasive procedures and to indicate that TURP is still competitive. PMID- 10828671 TI - Efficacy and cardiac safety of propiverine in elderly patients - a double-blind, placebo-controlled clinical study. AB - The study investigated the efficacy and cardiac safety of propiverine in the elderly, because the induction of life-threatening ventricular arrhythmia has been reported for some drugs prescribed in the therapy of urinary incontinence. Ninety-eight patients (21 male, 77 female; 67.7+/-6.3 years of age) suffering from urgency, urge incontinence or mixed urge-stress incontinence were included in the double-blind, multicentre, placebo-controlled, randomized study. After a 2 week placebo run-in period, the patients received propiverine (15 mg t.i.d.) or placebo (t.i.d.) for 4 weeks. Before (V1, V2) and during the treatment period (V3, V4), standard ECGs and 24-hour long-term ECGs were recorded. Propiverine caused a significant reduction of the micturition frequency (V2: 8.7+/-4.2, V4: 6.5+/-3.2 ml; p< or =0.01), reflected in a significant increase in the average micturition volume (V2: 163.5+/-65.9, V4: 216.3+/-101.5 ml; p< or =0.01) and a significant decrease in episodes of incontinence (-54%; p = 0.048). These findings were confirmed by the overall assessment at V4, in which approximately 90% of patients under propiverine either had no urge incontinence or urge symptoms, or showed improvement. Resting and ambulatory electrocardiograms indicated no significant changes. Neither the frequency-corrected Q-T interval nor other cardiac parameters were relevantly altered. The frequency of cardiac events (Lown classes IVa/b) was random, revealing no difference between placebo and propiverine. The incidence of adverse events was very low (2% dryness of the mouth under propiverine) and confirmed by the findings from the quality of life questionnaires. A favourable benefit-risk ratio without the induction of any cardiac arrhythmia in the treatment of elderly patients suffering from urgency, urge incontinence or combined urge-stress incontinence is therefore proven for propiverine. PMID- 10828672 TI - Low-dose oxybutynin for the treatment of urge incontinence: good efficacy and few side effects. AB - OBJECTIVE: Oxybutynin is an efficacious pharmacotherapeutic agent for the treatment of urge incontinence. However, many patients discontinue therapy because of the considerable side effects. This study aims at assessing the efficacy of a low-dose oxybutynin (2.5 mg three times daily) in relation to the concomitant profile of side effects. METHODS: Unselected primary care practice patients with involuntary urine loss were eligible for this study. A specially designed questionnaire was used to establish the diagnosis of symptomatic urge incontinence. All patients started with an oral dose of oxybutynin of 2.5 mg three times daily, and the responses with regard to efficacy and side effects were assessed after 2 and 6 weeks. Efficacy was evaluated using subjective patients' impression as well as data extracted from their voiding diaries. Adverse events possibly related to the use of oxybutynin were specifically asked for. Patients who experienced little or no symptomatic relief and who experienced no side effects were given a higher dose (5 mg three times daily). RESULTS: General practitioners recruited 416 patients with symptomatic urge incontinence. Increasing the dose to 5 mg three times was preferred by 115 patients (28%). After 6 weeks 393 patients reported a partial or complete symptomatic cure (positive responder rate 95%). The average number of micturitions per 24 h decreased from 9.9+/-(SD)3.7 to 6.0+/-2.5 (p<0.001), and the average number of episodes of urine loss per 24 h decreased from 6.0+/-3.6 to 1.7+/-1.8 (p<0.001). A total of 123 patients (30%) reported side effects attributable to the use of oxybutynin; 42 patients (10%) had to stop the medication because of the severity of these side effects. CONCLUSION: Successful pharmacological treatment of symptomatic urge incontinence can be given in a primary care setting using oxybutynin. Starting with a low dose of 2.5 three times daily, a subjective and objective efficacy can be achieved with fewer side effects than reported in the literature. PMID- 10828673 TI - Familial ureteral duplication and ureterocele: two sisters and their father. AB - We studied a family in which the father and his two daughters had ureteroceles involving the upper half of a duplex system. Our report gives additional evidence for the genetic background of ureteroceles. PMID- 10828674 TI - Functional bladder capacity as predictor of response to desmopressin and retention control training in monosymptomatic nocturnal enuresis. AB - OBJECTIVE: To evaluate the efficacy of intranasal desmopressin (DDAVP) and retention control training (RCT) for monosymptomatic nocturnal enuresis in childhood and to assess the predictive value of daytime functional bladder capacity for both methods. MATERIALS AND METHODS: A total of 114 children with monosymptomatic nocturnal enuresis, of whom 99 (86.8%) wetted the bed every night, were treated with 1 of the 2 methods: intranasal DDAVP in 54 and RCT in 60 subjects. RESULTS: Twenty-one of 54 patients (38.9%) and 14 of 60 patients (23.3%) in the DDAVP group and the RCT group, respectively, achieved strong improvement (p = 0.061). Forty-five of 54 (90.0%) in the DDAVP and 35 of 60 (58.3%) in the RCT group had a more than 50% decrease in wet nights (p = 0.004). In the DDAVP group, the functional bladder capacities at baseline in responders and nonresponders were 82+/-22% and 56+/-20% of the predicted bladder capacity for their age (p<0.001). In the RCT group, responders and nonresponders did not differ in functional bladder capacity at baseline. CONCLUSION: DDAVP treatment is more effective than RCT in decreasing the number of wet nights in childhood nocturnal enuresis, but not so effective in children with a low functional bladder capacity. Daytime functional bladder capacity is a valuable predictor of response to DDAVP, but not so to RCT. PMID- 10828675 TI - The hydraulic antireflux valve: a new technique for reimplanting dilated ureters. AB - PURPOSE: Wide ureters have a high risk of urinary reflux if they are implanted in the intestinal segment. Since 1978, we used the continence hydraulic valve as an antireflux device without staples. MATERIALS AND METHODS: A total of 40 patients, 20-65 years old (mean age 50), 36 with bilaterally severely dilated ureters and 4 with unilaterally dilated ureters underwent reconstructive surgery by intestinal segments with a hydraulic antireflux valve (HAV). RESULTS: Mean follow-up was 62 months (range 14-110); upper tract dilatation had improved or stabilized in all patients but one in whom bilateral dilatation occurred with HAV stenosis. All patients underwent follow-up loopogram studies which revealed 2 cases of reflux (4 ureterorenal units). In 1 of the 2 patients the urinary reflux occurred after dessusception (disinvagination) of the nipple. CONCLUSIONS: The HAV is a safe and reliable procedure in preventing reflux when implanting wide ureters into intestinal segments. This technique can be used in continent urinary diversions with intestinal pouches, and can also be used at the proximal level of an ileoureteroplasty. There is no need for staples. PMID- 10828676 TI - Methylglyoxal-induced apoptosis in human prostate carcinoma: potential modality for prostate cancer treatment. AB - OBJECTIVE: To examine the cellular effects of methylglyoxal (MG), a toxic physiological metabolite, on human prostatic cancer PC-3 cells. METHODS: The effects of MG on cell growth and viability were evaluated first, and then its effects on the cell cycle and the glycolytic process were analyzed by Western blots and specific assays. Possible MG-induced apoptosis was also assessed by DNA analysis using agarose gel electrophoresis. RESULTS: MG > or =3 mM caused severe growth inhibition, resulting in nearly 100% cell death by 24h. The time course study revealed that expression of cyclin D(1), cdk2, and cdk4 was significantly (>50%) downregulated in 3 h of MG (3 mM) exposure, followed by the dephosphorylation of retinoblastoma protein by 6 h. Both the glyceraldehyde-3 phosphate dehydrogenase activity and the cellular lactate level were also reduced by approximately 50 and 80%, respectively, following 6-hour MG exposure. Induction of apoptosis by MG was indicated by partial degradation of poly(ADP ribose) polymerase and further confirmed by discrete DNA fragmentation detected on an agarose gel. CONCLUSION: MG is capable of inducing apoptosis in prostatic cancer PC-3 cells, due primarily to a blocking of the cell cycle progression (G(1) arrest) and glycolytic pathway. Therefore, MG could be a potent apoptosis inducer, which may have a potential for prostate cancer treatment. PMID- 10828677 TI - Effect of beta-sitosterol on transforming growth factor-beta-1 expression and translocation protein kinase C alpha in human prostate stromal cells in vitro. AB - BACKGROUND: Today, plant extracts are widely used in the treatment of benign prostatic hyperplasia (BPH). However, the complete mode of action of the active substance, beta-sitosterol, is under investigation. The purpose of this study was to investigate the effect of beta-sitosterol on the expression of transforming growth factor beta 1 (TGF-beta1) and the activity of protein kinase C alpha (PKC alpha) in primary prostate stromal cell cultures in vitro. METHODS: Tissue samples for primary cell cultures were obtained from patients undergoing transurethral resection of the prostate (TURP). TGF-beta1 levels in stromal cell conditioned media following a culture with beta-sitosterol were detected in a TGF beta1 specific ELISA assay. Following different incubation periods with beta sitosterol, cells were lysed and fractionated into a Triton-soluble membrane fraction and a cytosol fraction. PKC-alpha protein was detected using immunoblot analysis. RESULTS: Beta-sitosterol was able to induce the expression and secretion of TGF-beta1 significantly between 1.26- and 1.86-fold compared to a cholesterol and the nonsupplemented control in 6 of 8 individual cultures. The total amount of secreted TGF-beta1 varied in cells from different patients. Based on its presence in both membrane fraction and cytosol, PKC-alpha appeared to be constitutively expressed in stromal cells. In the absence of beta-sitosterol PKC alpha was predominantly found in its membrane-associated active form. Following a culture with beta-sitosterol, a translocation of PKC-alpha from the membrane to the cytosol was observed. This effect was specific for beta-sitosterol as compared to cholesterol. CONCLUSION: This study describes the effect of beta sitosterol on the expression of a multifunctional growth factor (TGF-beta1) and the activity of PKC-alpha membrane in stromal cells of the human prostate in vitro. PMID- 10828678 TI - BPH: problems with terminology. PMID- 10828679 TI - Radiological evaluation of urethral disease PMID- 10828680 TI - Surgical management of urethral strictures PMID- 10828681 TI - Treatment of superficial bladder tumors: achievements and needs. The EORTC Genitourinary Group. AB - OBJECTIVE: The therapeutic objectives in the initial treatment of superficial tumors are to remove completely the tumor, to assess the need for further therapy and to plan the follow-up. METHODS/RESULTS: The EORTC Genitourinary Group assessed the percentage of patients with recurrence at 3 months (3RR) after complete resection of all visible lesions taking into account the institution, the number of tumors at presentation and the year of treatment. The 3RR was considered for 18 institutions. For single tumors, the 3RR varied from 0 to 36% and for multiple tumors from 7 to 75%. The 3RR by number of tumors was 8.7% for single tumors, 21% for 2-5 tumors and 32.2% for >5 tumors. The 3RR by year of entry for single tumors ranged from 21.0 to 43.8% during 1975-1978, from 6.3 to 12.7% during 1984-1986 and from 3 to 5.3% during 1987-1989. For multiple tumors it ranged from 50.0 to 61.5% during 1975-1978, from 20.2 to 27.3% during 1979 1983 and from 14.4 to 24.6% during 1984- 1986. The use of more refined instruments probably led to the decreasing percentage of the 3RR in more recent years, the large variation between institutions remains unexplained. The bladder's unique location renders its mucosa accessible to instillation of chemotherapeutic and immunotherapeutic agents. Cytostatics can be instilled into the bladder hours after surgery without severe complications. A single early instillation within 6 h after transurethral resection (TUR) in patients with a solitary bladder tumor category T(a)/T(1)G(1) to G(3) could reduce the recurrence rate per year by nearly 50%. The superiority of any of the commonly used intravesical drugs has never been demonstrated; the time to initiate therapy is important for treatment outcome. Optimal results can be achieved by initiating treatment early (within 24 h after TUR) and for a duration of 6 months, and maintenance (>6 months) for patients with a delayed first instillation (>7 days after TUR). Bacillus Calmette-Guerin (BCG) immunotherapy has been confirmed to be highly effective in the reduction of tumor recurrence, the treatment of residual papillary transitional cell carcinoma and the treatment of carcinoma in situ (CIS). The response rate in the treatment of the papillary disease averages 55%, and for CIS 73%. In the prevention of tumor recurrence the relative benefit of BCG is 45%. A direct prospective randomized comparison of BCG with intravesical chemotherapy has found it to be significantly superior to thiotepa, to doxorubicin and to mitomycin C when only patients with intermediate and high risk for recurrence were treated. In studies including patients with low recurrence risk, no advantage for BCG was found. Clinical trials showed no superiority of BCG immunotherapy to chemotherapy in preventing progression to > or =T(2). CONCLUSIONS: Investigation of the concept of chemoimmunotherapy up to now lacked evidence of advantages for this approach. Preventive regulatory measures directed to decrease tobacco smoking and some occupational exposures to aromatic amines may contribute to the reduction of bladder cancer. Bladder cancer is a multistep process making this tumor a candidate for chemoprevention. To date, retinoids are the best-studied chemopreventive agents achieving mixed clinical results in superficial bladder tumors. The potent apoptosis-inducing retinoid fenretinide is currently in the phase III trials. The follow-up of patients with all types of superficial tumors must be lifelong; unfortunately cystoscopy cannot be replaced yet by the control of any markers present or not in the urine. There is hope this may change in the near future. PMID- 10828682 TI - Biological response modifiers for the treatment of superficial bladder tumors. AB - BACKGROUND: For more than 20 years, superficial bladder tumors have been demonstrated to be sensitive to several biological response modifiers and especially to immunomodulators. The best-known and studied immunomodulator is the bacillus Calmette-Guerin (BCG). However, despite its well-recognized efficacy, BCG is not a universal panacea and is associated with potentially significant side effects. METHODS: New perspectives in BCG therapy aiming to increase BCG efficacy or to decrease side effects include the use of genetically engineered BCG strains producing cytokines as well as the use of purified BCG subcomponents. Because a cascade of immunological reactions including the secretion of several cytokines has been demonstrated in the BCG mode of action, many other biological response modifiers and especially immunomodulators have been studied for superficial transitional cell carcinoma therapy. Some were investigated in human trials, others are still in laboratory studies; some are administered intravesically whereas others are given orally. Interferon-alpha (IFN-alpha) intravesical instillations have been evaluated in several controlled studies. RESULTS: Although toxicity of intravesical IFN is minimal, its optimal dose, schedule and efficacy remain to be defined. Recent prospective studies comparing IFN to BCG intravesical therapy have been somewhat disappointing although this cytokine may be effective in some patients with T(a)-T(1) disease who have failed BCG therapy. Other immunomodulators administered intravesically investigated in clinical studies include interleukin 2 (recently used in a clinical study with a marker tumor response), levamisole, Rubratin, a Nocardia rubra cell wall skeleton, and keyhole limpet hemocyanin. Several biological response modifiers administered orally such as vitamin A (and its derivatives), Lactobacillus casei or bropirimine have been tested in clinical trials as well. In contrast, Allium sativum (garlic) or OK-432 (a streptococcal preparation) or BCG subfractions have been tested in laboratory studies only. CONCLUSIONS: Published reports on several of these biological response modifiers suggest that these compounds may be an alternative in patients with superficial bladder cancer who have failed or have not tolerated BCG, but further evaluation to improve efficacy, durability and understand their mechanism of action is warranted. PMID- 10828683 TI - Urinary cytokines reflecting the immunological response in the urinary bladder to biological response modifiers: their practical use. AB - BACKGROUND: Intravesical bacillus Calmette-Guerin (BCG) immunotherapy is currently the most effective treatment for superficial transitional cell carcinoma (TCC) of the urinary bladder. In recent years, the substantial number of patients not responding to BCG or experiencing considerable toxicities has stimulated studies addressing either the development of improved BCG treatment schedules or the exploration of the therapeutic value of a series of (novel) biological response modifiers, like interferons (IFNs), interleukin (IL) 2 and keyhole limpet hemocyanin. Although the actual mechanism by which BCG exerts its antitumor effect still needs detailed unraveling, current available knowledge suggests the induction of a T helper 1 (Th1) or Th1-like cytokine profile, represented by IL-2, IL-12 and IFN-gamma, as essential in the development of a cell-mediated antitumor activity. CONCLUSIONS: In this review, it is argued that incorporation of urinary cytokine determinations, like IL-2 and possibly IL-12 and IFN-gamma, may represent a valuable approach in the optimization and individualization of the BCG therapy and an early, initial evaluation of the potential efficacy of novel immunomodulating agents in the treatment of superficial TCC. PMID- 10828684 TI - Keyhole limpet hemocyanin: molecular structure of a potent marine immunoactivator. A review. AB - OBJECTIVES: In this short review we present a survey of the available biochemical and electron microscopic data on keyhole limpet hemocyanin (KLH). RESULTS: The biosynthesis of KLH and its biological role are discussed and the purification of the two isoforms of KLH (KLH1 and KLH2) presented in some detail. The determination of the molecular mass of KLH, its functional unit structure, carbohydrate content, immunological analysis and aspects of the molecular biology of KLH are all dealt with. Transmission electron microscopy (TEM) and crossed immunoelectrophoresis have played a significant part in the understanding of KLH structure. We present a summary of TEM studies on the native oligomers of KLH, the experimental manipulation of the different oligomeric states, immunological analysis and subunit reassociation. CONCLUSION: This fundamental structural information provides the scientific background upon which the understanding of the in vivo immunostimulatory function of KLH can be based. PMID- 10828685 TI - Keyhole limpet haemocyanin in experimental bladder cancer: literature review and own results. AB - OBJECTIVES: Keyhole limpet haemocyanin (KLH) is a high-molecular-weight protein antigen collected from the haemolymph of the sea mollusk Megathura crenulata. It is a powerful non-specific immune response modifier that induces both a cell mediated and a humoral response in animals and man. Thus, it is commonly used clinically as a measure of immune competence. In 1974, Olson studied the immune competence of bladder cancer patients by intradermal application of KLH. He later observed a significant reduction of recurrent disease in this patient group compared to another not immunized with KLH. This prompted a variety of experimental and clinical studies using KLH as an immunotherapy for recurrent bladder cancer. METHODS: Three different bladder cancer models have been used for experimental studies: intravesical transplantation of tumour cells in syngeneic mouse bladders; subcutaneous transplantation of tumour cells in syngeneic mice; direct chemical induction of bladder tumours by feeding rats with the carcinogen N-butyl-N-(4-hydroxybutyl)nitrosamine. RESULTS: The efficacy of KLH as an immunotherapeutic agent has been compared with different immune response modifiers alone or in combination with these in 11 experimental studies. Most of the studies used different concentration and application schedules for KLH. In addition a pre-immunisation prior to inoculation of the tumour was not performed in all studies. Therefore it is not useful to compare the results of these studies. However, most of the experiments demonstrated a significant effect on tumour appearance and extension after treatment with KLH. Intralesional or systemic application of KLH seemed to be superior to intravesical treatment. Pre immunisation with KLH several days or weeks before tumour inoculation also seems to be a key point of success. No study reported severe side-effects after application of KLH; additionally performed toxicity studies underlined the good tolerability of KLH. CONCLUSION: Based on all the experimental studies, KLH has to be judged as an effective and safe immunotherapeutic drug for the treatment of experimental bladder cancer. Prospective randomised clinical trials should evaluate the role of KLH as an immunotherapeutic alternative in the prophylaxis of recurrent bladder cancer and should determine whether the efficacy of KLH in man may be improved by systemic application. PMID- 10828686 TI - Keyhole limpet hemocyanin immunotherapy of bladder cancer: laboratory and clinical studies. AB - BACKGROUND: Since the serendipitous observation by Olsson in 1974 that patients immunized with 5 mg of keyhole limpet hemocyanin (KLH) had a marked reduction in recurrence of superficial bladder cancer, multiple laboratory and clinical studies have confirmed the efficacy of KLH immunotherapy. RESULTS: In 1981, we reported that KLH immunotherapy reduced tumor growth and prolonged survival in the MBT-2 murine model of transitional cell carcinoma (TCC), and in 1988, Jurincic and co-workers demonstrated that KLH was superior to mitomycin C chemotherapy in preventing bladder tumor recurrence. Subsequent studies using Immucothel (Biosyn), crude KLH, and endotoxin-free KLH confirmed the efficacy of KLH immunotherapy in the MBT-2 murine bladder cancer model (p < 0.05), and resulted in up to 100% survival. CONCLUSIONS: To evaluate the efficacy of KLH immunotherapy in patients, a multicenter clinical trial was performed. Sixty-four patients with CIS or residual stage T(a), T(1) TCC, or both were enrolled in a phase I-II trial of escalating doses of weekly KLH given intravesically for 6 weeks. Patients were followed with cystoscopic examination, urine cytology, and bladder biopsy. Complete response was seen in 50% of patients with CIS, 20% of patients with residual T(a), T(1) TCC, and 33% of patients with both CIS and residual T(a), T(1) TCC. Responses occurred at all doses tested: 0.4, 2, 10 and 50 mg. No significant difference in response according to dose was noted, but optimal overall complete response was seen with a dose of only 2 mg. The toxicity of KLH is minimal. KLH appears to be a safe and highly effective immunotherapy for superficial bladder cancer. PMID- 10828687 TI - Keyhole limpet hemocyanin for carcinoma in situ of the bladder: a long-term follow-up study. AB - OBJECTIVE: Keyhole limpet hemocyanin (KLH) is a nonspecific immunomodulator, demonstrated to be clinically effective in superficial bladder cancer. The present study investigated the clinical efficacy of intravesical KLH in patients with carcinoma in situ (CIS) with a long-term follow-up. METHODS: Thirteen patients with CIS grade III were treated with intravesical instillations of KLH, 20 mg for 6 weeks, then monthly for 1 year and bimonthly for 2 subsequent years. Patients not responding to 2 courses of KLH were treated with bacillus Calmete Guerin (BCG, 81 mg Connaught strain). RESULTS: The follow-up period ranged from 12 to 84 months. Two patients were free of tumor after KLH instillations with a follow-up of 66 and 82 months, respectively. All patients who did not respond to the primary KLH course, but to the 'rescue' instillation of BCG, experienced recurrences after 42, 48, 56 and 60 months after the first KLH instillation treatment. Three patients with recurrent CIS and who were not cystectomized had recurrences after prolonged remission (4-5 years). Patients progressing despite KLH and BCG instillations underwent cystectomy. CONCLUSIONS: KLH demonstrates efficacy and induces long- term remissions against CIS in a limited number of cases. In the present study, most patients with CIS progressed over time whatever the substance instilled, whether KLH or BCG. CIS remains a very aggressive neoplasm requiring a lifelong follow-up. Further studies are necessary to define the precise role of KLH in patients with CIS. PMID- 10828688 TI - Efficacy of prophylactic Immucothel in patients pretreated with conventional drugs to prevent recurrence of superficial bladder carcinoma. AB - OBJECTIVES: Fifty patients enrolled in clinical trials with Immucothel((R)) were reevaluated for their disease-free intervals. METHODS: Of the 37 evaluable patients, 16 had received mitomycin C, 3 bacillus Calmette-Guerin, 3 Adriamycin and 15 miscellaneous pretreatments prior to Immucothel. They thus served as their own controls. RESULTS: Although their prognosis was worse at the time when Immucothel therapy was started than at the time of initiation of pretreatment, the mean recurrence-free intervals of all patients increased from 17.0 months under pretreatment to 35.1 months under Immucothel. CONCLUSION: The difference was statistically significant (Wilcoxon matched-pairs signed-ranks test). PMID- 10828689 TI - Prevalence and etiology of linear enamel hypoplasia in monkeys and apes from Asia and Africa. AB - Ninety-seven specimens of sympatric monkeys and apes from East Malaysia and 115 monkeys and apes from West Africa are examined in order to evaluate the magnitude and nature of the great ape-monkey linear enamel hypoplasia (LEH) 'dichotomy'. This study demonstrates that great apes from both regions have a higher incidence of LEH and repetitive LEH than do gibbons and monkeys. However, the authors find that the dichotomy is not as clear-cut as previous research suggests, since some monkey samples exhibit high LEH frequencies. The authors evaluate the potential influence of great ape-monkey differences in crown height on this dichotomy. They show that canine crown height variation is weakly associated with LEH variation. Differences between monkeys and great apes in their crown formation spans and in their experience of environmental stress may be more likely causes of the dichotomy. PMID- 10828690 TI - Diet and feeding behaviour of the black-and-white ruffed lemur (Varecia variegata variegata) in the Betampona Reserve, eastern Madagascar. AB - The feeding behaviour and diet of the black-and-white ruffed lemur (Varecia variegata variegata) was investigated in the Betampona Reserve, eastern Madagascar. The highly frugivorous diet of this subspecies was confirmed - feeding on fruits accounting for 92.0% of feeding records. Most feeding at Betampona was observed at 10-25 m above the forest floor amongst flexible, small (0.5-5.0 cm diameter) and oblique/horizontal (0-45 degrees ) supports. The Varecia spent on average 21.7% (+/- 1.5) of their daily activity budget feeding and employ a variety of postures that enable them to harvest fruits in the rain forest canopy. The suspensory postures were the most important in allowing Varecia to compete with other smaller-bodied frugivores. PMID- 10828691 TI - Maternal recognition of infant calls in ring-tailed lemurs. PMID- 10828692 TI - Extragroup copulation among wild red howler monkeys in Venezuela. PMID- 10828693 TI - Swimming behaviour among bonnet macaques in Tamil Nadu. a preliminary description of a new phenomenon in India. PMID- 10828694 TI - Occurrence of twins in wild Formosan macaques, Macaca cyclopis, at Mt. Longevity, Taiwan. PMID- 10828695 TI - A survey of Tonkin snub-nosed monkeys (Rhinopithecus avunculus) in northern Vietnam. PMID- 10828697 TI - Two takeovers in wild hamadryas baboons. PMID- 10828696 TI - Multilocus DNA fingerprinting using oligonucleotide probes in 5 macaque species. PMID- 10828698 TI - Colour and shape discrimination in black lemurs (Eulemur macaco). PMID- 10828699 TI - Twenty new microsatellite loci for use with hair and faecal samples in the chimpanzee (Pan troglodytes troglodytes). PMID- 10828700 TI - Seasonal variation in food resource and forest strata use by brown capuchin monkeys (Cebus apella) in a disturbed forest fragment. PMID- 10828701 TI - Accurate multiplex polymerase chain reaction assay for gender determination from a single cell. AB - For gender determination of preimplantation embryos or circulating fetal cells in maternal blood, we developed a multiplex polymerase chain reaction assay from a single cell. This assay which co-amplifies X (DXZ1)- and Y (DYZ1)-specific repeat sequences, yields a 308-bp band in females and two bands of 154 and 308 bp in males. In a randomized, blinded assay of 100 isolated single amniocytes, 99 (99%) were amplified successfully. All 50 of the XY cells were correctly diagnosed as male (100%), whereas 49 of the 50 XX cells were diagnosed as female (98%). This accurate and efficient assay may be applicable in these clinical settings. PMID- 10828702 TI - Multiple screening for fetal Down's syndrome with the classic triple test, dimeric inhibin A and ultrasound. AB - The aim of the study was to determine whether inhibin combined with ultrasound improves the classic second trimester triple test. Eighteen Down's syndrome and 200 euploid pregnancies were included. Maternal serum alpha-fetoprotein (AFP), human chorionic gonadotrophin (hCG), and estriol (E3), measured between 15 and 20 weeks' gestation by radioimmunoassay, were retrieved. Inhibin A was tested with a specific two-monoclonal-antibody ELISA. Ultrasound was performed between 15 and 25 weeks' gestation: femur length, nuchal thickness and morphological abnormalities were assessed. Statistical analysis used a multivariate discriminant analysis. Except for the classic triple test parameters, only inhibin and the standard deviation score (SDS) for femur length allowed us to make a statistically significant distinction between euploid and Down's syndrome pregnancies. For a 5% false-positive rate, the classic triple test sensitivity was 61%. When E3 was replaced with inhibin A, the sensitivity increased to 77.8% and to 66.7% with SDS femur length. Combining hCG, AFP, SDS femur length and inhibin A gave a sensitivity of 66.7% for a false-positive rate of 2.5%. In conclusion, the classic triple test can be more significantly improved by replacing E3 with inhibin than by including ultrasound. However, a combination of ultrasound and inhibin enhances both the sensitivity and specificity of the test. In Down's syndrome screening, the biological parameters improve sensitivity while ultrasound is a better indication of specificity. PMID- 10828703 TI - Reproductive outcome after surgical treatment of ectopic pregnancy. AB - OBJECTIVES: Reproductive outcome after laparoscopy or laparotomy performed for treatment of ectopic pregnancy. METHODS: Data of 104 women were collected by a questionnaire and from medical records. RESULTS: No differences in pregnancy rate were found comparing the two different operative procedures. Women with a normal contralateral fallopian tube had a better pregnancy rate compared with those with a damaged contralateral tube. CONCLUSIONS: The crucial factor for future reproductive outcome after surgical treatment of ectopic pregnancy is the status of the contralateral tube. PMID- 10828704 TI - Comparison of magnesium and methyldopa for the control of blood pressure in pregnancies complicated with hypertension. AB - OBJECTIVES: Although magnesium is now the drug of choice for the prevention of eclamptic seizures only few studies have evaluated whether magnesium may reduce blood pressure in pregnancies complicated with hypertension. METHODS: A total of 33 patients with pregnancy-induced hypertension were randomized to either magnesium or methyldopa treatment. Of these 16 received magnesium and 17 methyldopa. The treatment comprised a 48-hour magnesium infusion followed by oral magnesium tablets until 3 days after delivery or 250 mg methyldopa 4 times a day in a similar period. RESULTS: Patients treated with magnesium had 1 day after inclusion a statistically significantly lower systolic blood pressure compared to the level in the methyldopa group (138.1 +/- 11 vs. 147.6 +/- 11 mm Hg; p < 0.05), but no difference was observed in diastolic blood pressure (92.0 +/- 6.6 vs. 96.0 +/- 10.1 mm Hg; NS). From the 5th day of inclusion and until delivery both systolic and diastolic blood pressure were significantly lower in the magnesium group (p < 0.05). Including all blood pressure measurements in a single analysis showed that both systolic (138 +/- 13 vs. 148 +/- 15 mm Hg; p < 0.0001) and diastolic (92 +/- 10 vs. 94 +/- 10 mm Hg; p < 0.05) blood pressure were lower in the magnesium group compared to the methyldopa group. There was no difference between the two groups regarding gestational age at delivery, birth weight, Apgar scores and pH in umbilical cord blood. CONCLUSION: This preliminary study demonstrates that magnesium treatment lowers blood pressure in pregnancies complicated with hypertension. The effect is without any adverse effect on maternal and neonatal well-being. PMID- 10828705 TI - Subclassification of small-for-gestational-age fetus using fetal Doppler velocimetry. AB - Our purpose was to determine whether small-for-gestational-age (SGA) fetus can be divided to subclassified groups using fetal Doppler velocimetry. Fifty-four pregnant women with SGA infant delivered after 37 weeks of gestation were studied. After 24 weeks of gestation, fetal middle cerebral artery puslatility index (MCAPI) and umbilical artery pulsatility index (UAPI) were measured at 2- to 3-week intervals using Doppler ultrasound. Perinatal outcomes [operative delivery due to fetal distress, abnormal fetal heart rate (FHR) pattern, meconium staining, low Apgar score (<7), neonatal acidosis (umbilical artery blood pH <7.15), neonatal intensive care unit (NICU) admission due to neonatal asphyxia, and decreased amniotic fluid] were compared in subclassified SGA groups using fetal Doppler velocimetry. The number of SGA fetuses with normal MCAPI and UAPI (normal SGA group) was 39, and those with significantly low MCAPI but normal UAPI (eventful SGA group) 15, respectively. Birth age and birth weights in the eventful SGA group were significantly earlier and lower than those in the normal SGA group, respectively (p < 0.05, and p < 0.005). There were significant increases in operative deliveries, abnormal FHR patterns and decreased amniotic fluid in eventful SGA group, when compared with events related to normal SGA group. However, there were no significant differences in meconium staining of amniotic fluid, low Apgar score, neonatal acidosis, and NICU admission between the two groups. These results suggest that SGA fetus with abnormally low MCAPI but normal UAPI has more poor perinatal outcomes, compared with that with normal MCAPI and UAPI. PMID- 10828706 TI - Sweeping of the membranes versus uterine stimulation by oxytocin in nulliparous women. A randomized controlled trial. AB - The aim of this study was to evaluate whether sweeping of the membranes at term could shorten the length of pregnancy and reduce the incidence of postterm pregnancies. We randomly selected 104 nulliparas with uncomplicated pregnancy and gestational age between 281 and 287 days. Our patients were divided into three groups. Group A consisted of 34 women who were subjected to sweeping of the membranes. Uterine stimulation with oxytocin was applied in 35 women (group B), and 35 women (group C) were used as a control group. We had no significant reduction of the time interval from sweeping of the membranes until delivery (1.9 +/- 1.2 days), compared to that of group B (2.1 +/- 0.8 days) as well as that of the control group (2.5 +/- 0.9 days). The incidence of spontaneous labor in patients after sweeping of the membranes was greater (67.6%) when compared with oxytocin-stimulated patients and the control group (p < 0.05). Furthermore, a better Bishop score was noted in patients of group A. No statistically significant difference was noted in the mode of delivery between the groups, but sweeping of the membranes significantly decreased the incidence of postterm pregnancies (p < 0. 05). We concluded that sweeping of the membranes is an effective method for initiating labor in women with a gestational age between 40 and 41 weeks, thus reducing the need for induction. PMID- 10828707 TI - Amniotic fluid index and labor length of pregnancies induced beyond 41 weeks of gestation with unfavorable cervix. AB - We evaluated if the response to prostaglandin E2 (PGE2) induction, in pregnancies completing 41 gestational weeks, is correlated to amniotic fluid index (AFI) values. A follow-up was performed from the time of the induction to the time of delivery of 63 pregnancies resulting in a spontaneous delivery showing unfavorable cervical examination at 41 weeks of gestation. This was induced by means of intracervical administration of PGE2 gel (Dinoprostone 0.5 mg). If the cervix was still unfavorable after 12 h, another gel administration followed. The number of PGE2 administration and AFI were both used as variables to correlate the time remaining before the delivery and the probability of delivery (Kaplan Meier and Cox algorithms). Gestational age, parity, neonatal weight, and APGAR 5' were used as covariates. A cut-off of AFI > 6 better discriminates two groups regarding the probability of delivery at paired hours from the beginning of the induction. A statistically significant difference was demonstrated in only those patients which did not deliver within 12 h (44 cases). Multivariate analysis (Cox regression) yielded an adjusted odds ratio associated to the probability of delivery of 0.47 (0.23-0.95, 95% CI, p value = 0.0354) for AFI < or = 6 vs. AFI > 6. PMID- 10828708 TI - Activation of Ca2+-activated K+ channels in human myometrium by nitric oxide. AB - To investigate the mechanism of human uterine smooth muscle relaxation, the activation of Ca2+-activated K+ channels in cultured myometrial cells obtained from human pregnant myometrium at term by nitric oxide was evaluated at the single cell level using the patch-clamp technique. The open probability of the K+ channel after the addition of 3 x 10(-3) M isosorbide dinitrate, a nitric oxide donor (0.116 +/- 0.048) was significantly higher than that before the addition (0.059 +/- 0.032; n = 9, p < 0.01). In myometrial cells pretreated with lipopolysaccharide, activation of K+ channels was also noted after the addition of L-arginine (10(-4) M; open probability increased from 0.179 +/- 0.076 to 0.380 +/- 0.105, n = 9, p < 0.01: 10(-3) M; open probability increased from 0.073 +/- 0.050 to 0.242 +/- 0.098, n = 12, p < 0.01). Either 10(-3) M N-nitro-L-arginine methyl-ester, an inhibitor of nitric oxide synthase, or 10(-6) M methylene blue, an inhibitor of guanylate cyclase, abolished activation of the K+ channel by 10( 3) M L-arginine in pretreated myometrial cells with lipopolysaccharide. Application of 10(-3) M L-arginine to the intracellular surface of an excised inside-out patch in the myometrial cells pretreated with lipopolysaccharide failed to increase Ca2+-activated K+ channel activity, suggesting that the activation was mediated by intracellular messengers. These results indicate that nitric oxide should control human myometrial relaxation during pregnancy via activation of Ca2+-activated K+ channels. PMID- 10828709 TI - Reproductive history in mothers of children with neural tube defects. AB - The reproductive history of 100 women with at least 1 child with a neural tube defect (NTD) has been studied. The data analyzed correspond to the period previous to their first visit to a genetic counseling service. A total of 204 pregnancies resulted in 205 outcomes. Of the 100 sibships, 14 (14%) had more than 1 affected member. The pregnancy was shorter than 28 weeks in 56/205 (27%) of the total outcomes. Of 104 evaluable previous outcomes, 34 corresponded to short pregnancies, positioned before an affected (23/60, 38%), a healthy (2/18, 11%), or an undiagnosed product (9/26, 35%). Short pregnancies subsequent to affected outcomes were also increased. The inter-gestational interval varied according to diagnosis: it was longer in the affected group than in the healthy one (0.1 > p > 0.05) and the subsequent intervals were shorter for the affected group (p < 0.05). An increased number of abortions adjacent to affected offspring and a changing fertility pattern, depending on the product diagnosis, point to an environmental etiological component in this high-risk NTD group of mothers. PMID- 10828710 TI - Use of chlorhexidine-releasing nylon fibres to reduce device-related uterine infections. AB - Implantation of intrauterine devices (IUDs) is associated with an increased incidence of uterine infection, probably as a result of vaginal bacteria entering the uterus at the time of insertion. To reduce the incidence of IUD-related infections, the incorporation of antimicrobial agents into the tail of the device was studied. Chlorhexidine was shown to be released from within nylon hollow fibres at a rate of approximately 114 microg x day(-1) for 10 days. This rate of release was sufficient to exhibit a biocidal effect on bacteria embedded within a mucus gel in vitro. When these devices were implanted transcervically into the guinea-pig uterus they significantly reduced the extent of uterine bacterial contamination within 24 h. These findings indicate that chlorhexidine-releasing devices are potentially useful in the reduction of device-related infections. PMID- 10828711 TI - Prognostic value of p53 expression in stage IB1 cervical carcinoma. AB - The purpose of this retrospective study was to evaluate the patterns of p53 expression in stage IB1 squamous cell carcinoma of the uterine cervix, to compare p53 expression with clinicopathological findings, and to assess its prognostic value. 27 patients with stage IB1 squamous cell carcinoma of the uterine cervix underwent abdominal radical hysterectomy and pelvic lymph node dissection. Expression of p53 was studied immunohistochemically. Overexpression of p53 was detected in 33.3% of the tumors, low expression was seen in 11.1%, and negative expression was found in 55.6%. Deep cervical stromal invasion (> or = 1/2) was found to be associated with the increased risk of lymph node metastases (odds ratio = 17.5). A significantly lower percentage of patients survived when p53 overexpression was observed (p = 0.0315). Univariate analysis revealed that tumor size (2-3.9 cm), lymph node metastasis, tumor invasion into parametria, tumor invasion into blood/lymph vessels, squamous cell carcinoma antigen (> or = 2 ng/ml), and p53 overexpression had a significantly lower recurrence-free survival rate. None of these above factors obtained significance in the multivariate analysis. This study suggests that expression of p53 may be indicative of an unfavorable prognosis in patients with stage IB1 squamous cell carcinoma of the uterine cervix. PMID- 10828712 TI - Significance of serum and peritoneal fluid lactate dehydrogenase levels in ovarian cancer. AB - We investigated prospectively whether the detection of serum lactate dehydrogenase (LDH) and/or peritoneal fluid LDH levels may serve as a reliable biochemical marker in discriminating ovarian carcinoma from benign ovarian tumors. In this series, postoperatively 20 of 50 patients had a diagnosis of ovarian cancer while the remaining 30 patients had benign ovarian tumor. No significant difference in peritoneal fluid LDH levels was observed between patients with ovarian cancer and benign ovarian tumor (p > 0.05). Serum LDH levels in ovarian cancer patients were significantly higher than those in patients with benign ovarian tumor (p < 0.05). Statistically significant differences were not observed in LDH levels of different histological types of ovarian cancer and different stages of the disease. Serum LDH levels presented diagnostic accuracy with high specificity and may have a potential use as a biochemical marker. PMID- 10828713 TI - Congenital fiber-type disproportion presenting antenatally with clubfoot and hydramnios. AB - This is a case of congenital fiber-type disproportion that presented prenatally with bilateral clubfoot, hydramnios, and reduced fetal movements. Although prognosis is generally good for this condition, the neonate presenting at birth may have a more severe form of congenital fiber-type disproportion with a high rate of mortality. PMID- 10828714 TI - Coexistence of a heterotopic pregnancy associated with a homolateral ovarian cyst in a patient submitted to elective abortion. AB - The authors describe the case of a right tubal pregnancy of delayed diagnosis in a 31-year-old nullipara, who was submitted to voluntary termination during the 7th week of pregnancy and who presented a homolateral ovarian cyst. Two weeks later the patient presented pelvic pain and intraperitoneal fluid layer, while plasma beta-hCG was 1,262 IU/ml. The case history was complicated by recent termination surgery and presence of an ovarian cyst, but a plasma beta-hCG assay and transvaginal ultrasonography oriented the diagnosis towards a previously unrevealed heterotopic pregnancy. The fallopian tube and the ovarian cyst were removed by laparoscopy. The case points out to the fact that, though rare, heterotopic pregnancy must always be considered one of the possible complications of spontaneous pregnancy. PMID- 10828715 TI - Leiomyoma of the fallopian tube. AB - Leiomyomas of the fallopian tube are rare. They are typically incidental findings seen at autopsy or unrelated surgical procedures. A 32-year-old woman presented with lower abdominal pain and mass. Transvaginal sonogram and magnetic resonance imaging showed the solid mass at the outside of the uterus. At surgery, the left fallopian tube contained a firm mass with torsion in the area of the ampullary isthmic junction. The left tube and the infundibulopelvic ligament were rolled in torsion and showed edematous change. We report a rare case in whom torsion of a pedunculated tubal leiomyoma caused abdominal pain. PMID- 10828716 TI - Regulation of specific immune responses by chemical and structural modifications of allergens. AB - Specific immunotherapy (SIT) is an efficient treatment of allergic diseases to defined allergens. Despite being used in clinical practice since early in this century, more rational and safer regimens are required, because SIT is faced with the risk of anaphylaxis and standardization problems of allergen-extract-based treatments. A better understanding of the pathogenesis of allergy and of the mechanisms of SIT has led to various approaches to overcome these problems. Knowledge of the influence of IgE-facilitated antigen presentation on allergen specific Th2 responses increased the efforts to generate non-IgE-binding allergens. The current principal approach to allergen modification is to modify B cell epitopes in order to prevent IgE binding and effector cell cross-linking while preserving T cell epitopes to retain the capacity of inducing tolerance. In this way, the modified allergen will be directed to T cells by a phagocytosis/pinocytosis-mediated antigen uptake mechanism, bypassing IgE cross linking and IgE-dependent antigen presentation. Accordingly, a differential regulation of allergen-specific T cell cytokine patterns and IgE:IgG production was demonstrated by modifications of the three-dimensional structure of allergens because of linearity in T cell epitopes and conformation dependence in B cell epitopes. In this context, chemically modified allergen extracts with low IgE binding capacity have been developed to reduce anaphylactic side effects since the early 1980s. The progress of recombinant techniques for producing allergens and allergen derivatives has led to a dramatic improvement in the ability of developing novel vaccines for the treatment of allergy. This has enabled mutation or deletion of decisive amino acids in B cell epitopes and fractionation or oligomerization of allergens by genetic engineering as fruitful approaches to generate hypoallergenic vaccines. Moreover, non-IgE-binding short T cell epitope peptides and single-amino-acid-altered peptide ligands represent potential candidates for future SIT. PMID- 10828717 TI - Allergy and the gut. AB - There have frequently been doubts as to the relevance of food allergy, in particular as far as the involvement of the intestinal tract is concerned. Several studies, however, have confirmed the existence of allergic reactions in the gut, with an estimated prevalence of about 1-2% in adults. Clinical symptoms are unspecific and include nausea, vomiting, abdominal pain, cramping and diarrhea. Intestinal mast cells, as well as intestinal eosinophils, have been shown to be involved in the pathogenesis of food-allergy-related enteropathy. In addition to classical IgE-dependent degranulation, further agonists have been demonstrated for mast cell activation, for example IL-4. The methods used to confirm the diagnosis of intestinal allergy are still insufficient. Until now, blinded oral challenge procedures with food antigens have been accepted as the 'gold standard' in diagnosing food allergy, although these tests have practical problems. Therefore, new test systems have been developed, such as endoscopic provocation tests, that may improve diagnostic procedures. Elimination diet still presents the main basis of therapy. Aspects to be focused on in the future are the role fo IgE-independent allergic mechanisms in intestinal allergy, the impact of cross-reactivity with other allergens and the relationship to other inflammatory bowel diseases such as Crohn's disease, ulcerative colitis, celiac disease and irritable bowel syndrome. PMID- 10828718 TI - Purification and immunobiochemical characterization of folding variants of the recombinant major wasp allergen Ves v 5 (antigen 5). AB - BACKGROUND: Antigen 5 is one of three major allergens in wasp venoms, but unlike phospholipase A(1) and hyaluronidase, both of which are enzymes, its biological function is unknown. The cDNA coding for this allergen has been isolated and used for recombinant expression. Thorough analysis of the expression product is essential in order to evaluate the usefulness for in vivo or in vitro application. OBJECTIVE: In this study, folding variants of the recombinant major allergen Ves v 5 from Vespula vulgaris were immunologically and biochemically investigated in order to determine their possible applicability for diagnostic or therapeutic purposes. METHOD: The cDNA encoding Ves v 5 was cloned into the expression vector pSE420 which generates recombinant products lacking a tag sequence. After expression, inclusion bodies were purified, subsequently denatured and dialyzed against different solutions. The structural properties of soluble proteins were analyzed by size exclusion chromatography, non-reducing SDS PAGE, native PAGE, N-terminal sequencing, proteolytic digestion and ion exchange chromatography. Immunological investigations were performed by using different monoclonal antibodies (mAbs) specific for Ves v 5 and IgE from patients allergic to wasp venom allergens. RESULTS: After dialysis, soluble monomeric recombinant Ves v 5 was more than 95% pure in each case. Using different dialysis solutions, clearly distinguishable folding variants were obtained. In one case, the recombinant allergen was comparable with the natural counterpart in respect of migration in non-reducing SDS-PAGE, native PAGE and IgE reactivity. This variant reacted with two different Ves v 5-specific mAbs and produced a stable fragment after proteolytic digestion. Elution from a cation exchange chromatography column was achieved with 320 mM NaCl. In two other cases, folding variants exhibited a different migration behavior in SDS-PAGE and native PAGE compared with the natural allergen. Also, the mAb 1E11 recognized none of these variants since it presumably detected a conformational epitope. Moreover, the IgE reactivity was clearly reduced and proteolytic digestion effected almost complete degradation. These variants eluted from the cation exchange column with 400 mM NaCl. CONCLUSION: Defined folding strategies resulted in both soluble misfolded variants with reduced IgE reactivity, potentially suitable for immunotherapy, and natural-like folded variants for diagnosis. PMID- 10828719 TI - Skin prick test reactivity to recombinant latex allergens. AB - BACKGROUND: Allergy to latex has become a serious and increasingly common health problem, particularly for healthcare workers and patients who undergo frequent surgical procedures. Testing for latex allergy currently involves in vitro tests and skin prick testing using crude preparations of natural rubber latex (NRL). To date, 10 latex proteins have received designation as allergens (Hev b 1 to Hev b 10) and, except for Hev b 4, have been cloned as recombinant proteins. Our aim was to compare the skin prick test (SPT) reactivity of six recombinant latex allergens with SPT reactivity to natural rubber latex proteins in known latex allergic individuals. METHODS: Six recombinant proteins were expressed in Escherichia coli, and tested as the intact fusion proteins (Hev b 2, 5, 6, 8) or as purified proteins (Hev b 3 and 7). SPT with the six recombinant latex allergens was performed using 10-fold serial dilutions on 31 latex-allergic subjects to determine the level of reactivity to each recombinant allergen. Latex specific IgE was determined using the AlaSTAT assay. RESULTS: All six recombinant allergens were reactive by SPT in at least 1 latex-allergic patient but not in any of the control patients. The frequency of sensitization to the various recombinant allergens was similar to previous studies using the native proteins isolated from NRL. The minimal level of protein for a positive skin test was 70 pg/ml for NRL and 1 ng/ml for one recombinant allergen (Hev b 7). In our patients, the use of a combination of recombinant latex allergens Hev b 5, 6 and 7 diagnosed latex allergy with 93% sensitivity and 100% specificity. CONCLUSION: Recombinant latex allergens are clinically reactive, can be produced in a standardized manner, and could potentially provide safe, sensitive and specific reagents for the diagnosis of latex allergy. PMID- 10828720 TI - IgE-mediated sensitization to mould allergens among patients with allergic respiratory diseases in a desert environment. AB - BACKGROUND: The importance of fungal allergens in the development of allergic diseases in a desert environment is uncertain. This study evaluated the prevalence of IgE sensitization to moulds among patients with allergic respiratory diseases in Kuwait - a desert country. METHODS: A total of 810 patients (male:female ratio 1.4) with a mean age of 32.3 years (range 2-76 years) with extrinsic asthma or allergic rhinitis were studied. Sera from the patients were tested by the CAP-RAST method for specific IgE to 6 fungi (Penicillium, Cladosporium, Aspergillus, Candida, Alternaria and Helminthosporium). For comparison house dust mite and Bermuda grass were also assessed. RESULTS: The overall positivity to at least one mould was 20.9%. Among 120 matched control subjects, the value was 5. 8%. The value was much higher among patients with asthma alone (45. 8%) or both asthma and rhinitis (28.3%) than those with rhinitis alone (11.8%; p < 0.001). Asthmatic children had the highest sensitization rate (66.0% in the 7- to 12-year age group), which declined sharply with age. Among asthmatics, Candida and Aspergillus had the highest sensitization rates (23.1 and 21.3%, respectively), followed by Helminthosporium (18.8%), Cladosporium (15.9%), Alternaria (14.6%) and Penicillium (13.9%). The values for mite and Bermuda grass were 41.2 and 54.6%, respectively. Among asthmatic children, severe asthma was significantly more frequent among mould-positive (51.6%) than mould-negative patients (17.5%; p < 0. 0001). CONCLUSIONS: Even in this desert environment, sensitization to moulds is quite common among patients with allergic respiratory diseases, with a striking preponderance among children with asthma. Mould allergy could also be an important factor determining asthma severity in this environment. PMID- 10828722 TI - Enhancing effect of dietary oil emulsions on immune responses to protein antigens fed to mice. AB - Repeated intragastric administration of beta-lactoglobulin (beta-Lg) with emulsified soybean oil elicited an antigen-specific, systemic humoral immune response in different strains of mice. The antibody response was enhanced as the dose of oil was increased and the particle size of emulsions was decreased. Feeding of aqueous beta-Lg could induce the antibody response only when emulsified oil was fed almost simultaneously. However, the emulsion-driven humoral immune response was not observed when mice were treated with anti-CD40 ligand antibody or in athymic mice. It is likely that the intestinal coexistence of emulsified oil with dietary antigens modulates the immune system to crucially support B cell response. A practical application of the present results to the prevention of cow's milk protein sensitization in infants is proposed. PMID- 10828721 TI - Modulation of allergen-specific immune responses to the major shrimp allergen, tropomyosin, by specific targeting to scavenger receptors on macrophages. AB - BACKGROUND: Tropomyosin from shrimp is the major cross-reacting crustacean food allergen. Earlier studies have led to the purification and immunochemical characterization of the major IgE binding epitopes of the allergen. Maleylated proteins are known to be specifically targeted to scavenger receptors on macrophage. Since antigens processed and presented by macrophages are known to elicit Th1 type of responses and allergic responses are characterized by polarization towards Th2 phenotype, the possibility of modulation of allergen specific immune responses by targeting of tropomyosin to macrophage via scavenger receptor was explored. METHODS: The IgG and IgE binding potential of the native maleylated form of tropomyosin was carried out by ELISA and immunoblot. The ability of the native and maleylated form of allergen to induce in vitro proliferation of splenocytes from BALB/C mice immunized with both forms of allergen was tested. The in vitro production of IL-4 and IFN-gamma by splenocytes from mice immunized with the two forms of allergen was determined from culture supernatants. The in vivo production of serum IgG1 and IgG2a antibodies following immunization with native and modified allergens was monitored by ELISA. RESULTS: The maleylated form of tropomyosin was found to have reduced antigenicity and allergenicity as compared to its native counterpart. The modified allergen was, however, found to elicit a cellular response similar to native tropomyosin in vitro. Analysis of the cytokine profiles showed a modulation from an IL-4 dominant, proallergic, Th2 phenotype to an IFN-gamma-dominant, antiallergic, Th1 phenotype that could also be correlated to a modulation in the in vivo antibody isotype. CONCLUSION: The results suggest the possible potential for modulating allergic responses in vivo by selective targeting to macrophages. PMID- 10828723 TI - Soluble CD30 and cyclosporine in severe atopic dermatitis. AB - BACKGROUND: Allergen-reactive Th2-like cells expressing membrane CD30 are present in the circulation of atopic dermatitis (AD) patients during seasonal allergen exposure. Moreover, CD30+ T cells are present in the lesional skin of AD patients and high levels of soluble CD30 (sCD30) are found in the serum of the same atopic patients. To investigate the immunosuppressive capacity of cyclosporin A (CsA) in AD patients, the sCD30 serum level was determined before and after CsA treatment (5 mg/kg/day) in 10 patients with severe, refractory AD. The sCD30 serum levels before and after CsA therapy together with other serum parameters were correlated with disease activity. METHODS: sCD30 serum levels were detected using a commercial sandwich ELISA; serum eosinophil cationic protein (ECP) levels were determined using a radioimmunoassay (RIA). RESULTS: In all AD patients sCD30 serum levels were increased ranging from 36 to 300 U/ml, with a mean value equal to 135.7 U/ml. After 6 weeks of CsA treatment, not only was there a significant difference between serum sCD30 levels before (mean 135.7) and after (mean 96.2) treatment but even the serum ECP levels before (mean 57.78) and after (mean 18.69) therapy showed an important reduction. Moreover, no significant difference was found between the mean of serum IgE levels before and after treatment, although the values showed a correlation (p = 0.0003). No significant correlations could be demonstrated between sCD30 levels and serum IgE or between sCD30 and ECP serum levels nor between sCD30 levels and blood eosinophil count after CsA treatment. Moreover, a positive correlation (p = 0.001) was instead documented between sCD30 and the severity of the disease. CONCLUSIONS: In this study, CsA therapy results in clinical improvement together with a statistically significant reduction in sCD30 and ECP serum levels in AD patients. PMID- 10828724 TI - IgG immune complex binding to and activation of liver cells. An in vitro study with IgG immune complexes, Kupffer cells, sinusoidal endothelial cells and hepatocytes. AB - BACKGROUND/AIMS: The aim was to study IgG immune complex (IC) binding to isolated hepatocytes, Kupffer cells (KCs) and sinusoidal endothelial cells (SECs). Further, we wished to analyze the capacity of IgG ICs to induce release of reactive oxygen metabolites by the IC-binding liver cells. METHODS: ICs were formed between (125)I-tyramine-cellobiose-labelled dinitrophenyl-conjugated human serum albumin ((125)I-TC-DNP(10)HSA) and polyclonal rabbit IgG antibodies. Binding of ICs to different rat liver cells in suspension was studied at 4 degrees C. Production of reactive oxygen metabolites was measured by luminol enhanced chemiluminescence at 37 degrees C. RESULTS: IgG mediated binding of (125)I-TC-DNP(10)HSA to both KCs and SECs, but not to hepatocytes. The binding showed saturation kinetics and was blocked by an excess of unlabelled IgG-ICs. IgG-ICs activated KCs, but not SECs, to a chemiluminescence response. CONCLUSIONS: Both KCs and SECs bind IgG-ICs in vitro, probably via Fc receptor interaction. IgG-ICs activate KCs to produce reactive oxygen metabolites. The binding of IgG-ICs to isolated hepatocytes is small. PMID- 10828725 TI - Increased inhibitory activity of protein kinase C on the serotonin transporter in OCD. AB - Different observations show a reduced functionality of the serotonin (5-HT) transporter in obsessive-compulsive disorder (OCD) that might be due to a disturbance of its regulation at intracellular level. Protein kinase C (PKC) has been reported to provoke a decrease in the number of the 5-HT transporter proteins. Therefore, we investigated whether OCD patients differed from control subjects in the effect of PKC upon the 5-HT transporter, after stimulation of this enzyme with 4beta-12-tetradecanoylphorbol 13-acetate (beta-TPA). Fifteen patients affected by OCD, according to DSM-IV criteria, were compared with a similar group of healthy subjects. The determination of 5-HT uptake was carried out according to the method of Arora and Meltzer with slight modifications. At baseline, OCD patients showed a significant decrease in the maximal velocity (V(max)) of 5-HT uptake, as compared with control subjects, with no change in the Michaelis-Menten constant (K(m)). The activation of PKC with beta-TPA provoked a significant decrease in V(max) values in both groups, but the effect was significantly more robust in OCD patients who, in turn, also showed also an increase in K(m) values. These findings could indicate the presence of hyperactivity of PKC in OCD that could be the result of increased activity of the phosphatidylinositol pathway. In addition, this suggests new potential therapeutic targets in OCD. PMID- 10828726 TI - Effect of time-of-day on the yawning response to apomorphine in normal subjects. AB - The yawning response to the dopamine (DA) receptor agonist apomorphine HCl (Apo, 7 microg/kg s.c.) and placebo (physiological saline) were examined in two groups of normal men. One group (n = 11) was investigated in the morning and the other group (n = 16) in the afternoon. The frequency of yawning was polygraphically monitored for 60 min following injection. Apo increased yawning compared with placebo when given in the morning (p < 0.02), but not when given in the afternoon. Yawning frequency was increased after both Apo (p < 0.01) and placebo (p < 0.025) when given in the morning compared with responses in the afternoon. These results suggest that yawning frequency with both Apo and placebo is influenced by time of day, possibly as a result of diurnal variation in DA receptor sensitivity. PMID- 10828727 TI - Sleep laboratory studies in restless legs syndrome patients as compared with normals and acute effects of ropinirole. 1. Findings on objective and subjective sleep and awakening quality. AB - Although the restless legs syndrome (RLS) is a disorder with a relatively high prevalence rate (8% in Austria) and leads to insomnia and excessive daytime tiredness, there is a paucity of sleep laboratory data concerning objective and subjective sleep and awakening quality. Thus, the aim of this study was to investigate 12 untreated RLS patients as compared with 12 normal controls and subsequently measure the acute effects of 0.5 mg ropinirole (Requip((R))) - a nonergoline dopamine agonist - as compared with placebo. In 3 nights (adaptation, placebo, ropinirole night) sleep induction, maintenance and architecture were measured objectively by polysomnography, subjective sleep and awakening quality were assessed by self-rating scales and visual-analog scales, and objective awakening quality was evaluated by a psychometric test battery. In polysomnography, RLS patients demonstrated, as compared with normal controls, a decreased total sleep time (TST) and sleep efficacy, increased wakefulness during the total sleep period and frequency of nocturnal awakenings, increased sleep stage S1, decreased S2 and increased stage shifts. Subjective sleep quality tended to decrease, and morning well-being, mood, affectivity and wakefulness were deteriorated. In the noopsyche, fine motor activity and reaction time performance were deteriorated. Ropinirole 0.5 mg induced, as compared with placebo, an increase in TST, sleep efficacy, S2 sleep and stage shifts. In the morning, somatic complaints increased slightly, while fine motor activity and reaction time performance improved. Our findings suggest a key-lock principle in the diagnosis/treatment of RLS and a dopaminergic mechanism in its pathogenesis, which is supported by the data on periodic leg movements during sleep and arousals of the subsequent paper. PMID- 10828728 TI - Sleep laboratory studies in restless legs syndrome patients as compared with normals and acute effects of ropinirole. 2. Findings on periodic leg movements, arousals and respiratory variables. AB - The restless legs syndrome (RLS) is a common sensorimotor disorder which leads to severe sleep disturbances and showed a prevalence of 7.9% in our sleep laboratory. The aim of this study was to investigate periodic leg movements (PLM), arousal and respiratory variables in 12 untreated RLS patients and to measure the acute effects of 0.5 mg ropinirole, a nonergoline dopamine agonist, as compared with placebo. In the target variable PLM/h of total sleep time (PLM/h TST), RLS patients showed an increased value of 40/h (normal 0-5/h). Further, we found an increased number of PLM (368), PLM/h of time in bed (49/h), PLM/h of REM sleep (11), PLM/h of non-REM sleep (46) and PLM/h awake (61). The arousal index was also increased (32/h; normal 0-25/h), as were arousals due to PLM. In the confirmatory part of our descriptive data analysis, ropinirole 0.5 mg significantly improved, as compared with placebo, the index PLM/h TST by 75%. In the descriptive part, all the other PLM variables were improved as well. Arousals due to PLM decreased, while spontaneous arousals increased. Respiratory variables, which had a priori been in the normal range, showed a slight but significant improvement after the dopamine agonist. Thus, 0.5 mg ropinirole significantly improved the target variable PLM/h TST, along with objective and subjective sleep quality and morning noopsychic performance, as described in the preceding paper. Our data encourage further sleep studies including all above mentioned variables in a larger group of RLS/PLM during sleep patients as well as long-term efficacy trials. PMID- 10828729 TI - Determinants of the voluntary consumption of nicotine by rats. AB - The 2-bottle free-choice method was used to study the voluntary consumption of nicotine by rats. Rats consumed nicotine voluntarily at different, albeit quite consistent, amounts. Voluntary intakes were higher in younger than older rats, but were not affected by gender. A previously forced nicotine intake had no effect on a subsequent voluntary intake of nicotine in older but increased it in younger rats. Forced exposure to nicotine of pregnant and lactating rats did not increase the voluntary intake by their offsprings. Established free-choice drinking patterns of nicotine were not affected by a temporary forced intake of this substance. The 2-bottle choice proved to be a reliable method to study the voluntary intake of nicotine, and results suggest that nicotine is not 'addictive' per se, but that its use is apparently determined by the response of an individual rat to this substance. PMID- 10828730 TI - Influence of various drugs on the voluntary intake of nicotine by rats. AB - The effects of the dopaminergic agonists (L-dopa, pergolide) and antagonists (haloperidol, clozapin) and a cholinergic agonist (tacrine) and antagonist (mecamylamine) on the voluntary intake of nicotine were investigated with the 2 bottle paradigm with the test drugs being dissolved directly in the drinking fluid of the animals. This method was found to be a reliable procedure to quickly screen compounds with specific sites of action in the brain for their effects on the voluntary intake of nicotine or perhaps other substances of abuse as well. L dopa, pergolide and haloperidol did not affect the intake of nicotine, whereas tacrine increased it slightly and clozapine and mecamylamine markedly. These results indicate that blockade of nicotinic and dopaminergic D4 receptors partially reduce the desired effect of nicotine by forcing the animals to consume more of this substance. PMID- 10828731 TI - Acute nicotine administration in Alzheimer's disease: an exploratory EEG study. AB - Previous findings of cognitive deficits and EEG slowing in Alzheimer's patients, together with independent reports of the performance enhancing and electrocortical activating properties of nicotine in normal adults, stimulated this study to examine the acute effects of nicotine on spectrum-analyzed EEG in patients with dementia of the Alzheimer type (DAT). Thirteen patients, 6 currently receiving cholinesterase inhibitor treatment and the remaining being medication free, were administered 2 mg of nicotine polacrilex under randomized, placebo-controlled conditions. Compared to age-regressed EEG norms, the pretreatment EEG spectrums of patients in general were characterized by excessive slow (delta and theta)-wave power, diminished fast (alpha and beta)-wave power and slow mean alpha and total band frequencies. Although postnicotine EEG indices remained within the abnormal range, nicotine, compared to placebo, significantly shifted EEG towards normal values by reducing slow wave (relative delta and theta) power and augmenting fast (relative alpha-1, alpha-2, beta-1) wave power. No differences were observed between treated and nontreated patients in response to nicotine. The results are discussed in relation to cholinergic and brain arousal systems and their relationship to cognitive processes. PMID- 10828732 TI - F(2) isoprostanes, potential specific markers of oxidative damage in human retina. AB - F(2) isoprostanes (F(2)-IPs) are a series of prostaglandin-F(2)(PGF(2))-like compounds produced during peroxidation of arachidonic acid (AA) by a mechanism independent of the cyclo-oxygenase. Of these, 8-epi-PGF(2alpha) has proved a reliable marker of oxidative stress in studies both in vitro and in vivo. Human retinas (n = 28) were obtained from healthy donors (age 15-59 years) and analysed for the content of free (non-esterified) as well as total (sum of free and esterified) F(2)-IPs. Retina tissue was homogenised, total lipids were partitioned with ethyl acetate and isoprostanes were isolated by chromatography on an NH(2) cartridge. After formation of pentafluorobenzyl ester and trimethylsilyl derivatives, F(2)-IPs were analysed by negative ion-chemical ionisation mass spectrometry using tetradeuterated PGF(2alpha) as the internal standard. Analysis of retina samples revealed the presence of measurable amounts of esterified 8-epi-PGF(2alpha) (mean: 81.71 microg/g AA; range: 3.29-187.89 microg/g AA) but not free F(2)-IPs. There was no correlation between age and content of 8-epi-PGF(2alpha). Detection of esterified F(2)-IPs in human retina provides the first direct evidence for free-radical-catalysed peroxidation of AA in retinal tissue. Analysis of F(2)-IPs should provide new opportunities for studying the implications of oxidative injury in various diseases of retina, including diabetic retinopathy and age-related macular degeneration. PMID- 10828733 TI - Nitric oxide as a second messenger in phagocytosis by cultured retinal pigment epithelial cells. AB - PURPOSE: To investigate a possible role of the nitric oxide (NO)-cGMP signal transduction system in phagocytosis of rod outer segments (ROS) by cultured retinal pigment epithelial (RPE) cells. METHODS: Primary cultures of RPE cells from 10-day-old Brown Norway rats were used to study the phagocytosis of ROS by these cells. Phagocytosis of ROS was evaluated with or without an inhibitor of nitric oxide synthase (NOS), N(G)-nitro-L-arginine (L-NNA), and the reverse effects of L-NNA by L-arginine and 8-bromo-cGMP on phagocytosis were also studied. NO-associated cGMP production by RPE cells was monitored during phagocytosis using L-NNA. NOS activity was assayed in RPE cells and ROS to locate the source of NO. RESULTS: Phagocytosis of ROS was inhibited by L-NNA but not by D-NNA. L-NNA inhibited the ingestion in a dose-dependent manner, but not the binding of ROS. The inhibition was reversed by L-arginine and also by an NO donor, SIN-1. RPE cells challenged with ROS showed increased cGMP activity, which was significantly reduced by L-NNA and again restored by an overdose of L arginine. NOS activity was found in RPE cells but not in ROS. CONCLUSIONS: Our data show that cGMP plays a role in the ingestion phase of ROS phagocytosis by RPE cells via a cGMP second-messenger system. PMID- 10828734 TI - The effects of induced acute hyperglycemia in the cat on the retinal capillary blood flow. AB - PURPOSE: To study the effect of acute hyperglycemia on the erythrocyte flow in specific retinal capillary paths. METHODS: A technique for fluorescent labeling of a known fraction of the erythrocyte moiety of systemic blood was combined with fluorescence viewing of the retinal capillary network in live cats. This technique was developed to enable visualization of the erythrocyte flow in the retinal capillary network and used to acquire video recording of the retinal capillary erythrocyte flow in normal feline eyes. The pattern of capillary erythrocyte flow under normal blood glucose levels and normal systemic blood pressure served as baseline. Acute hyperglycemia was induced by intravenous injection of glucose while monitoring the systemic blood pressure. Two subsets of functional capillary pathways previously defined as S (simple) and C (complex) were identified in the recorded data. The relationship between erythrocyte flux in random selections of these two pathways and the level of hyperglycemia and systemic blood pressure was determined. RESULTS: Induction of acute hyperglycemia led to acute elevation of the systemic blood pressure that returned to baseline levels within few minutes, while blood glucose remained high. The capillary erythrocyte flux in S paths was significantly higher than in C paths at all values of systemic blood pressure. The capillary erythrocyte flux in the S paths was directly proportional to the systemic blood pressure whereas the flux in C paths was minimally affected by acute hyperglycemia for the systemic blood pressure range between 110 and 160 mm Hg. CONCLUSIONS: The erythrocyte flux in S paths is affected by the changes of the systemic blood pressure and these paths act as 'shunt vessels' when acute elevation of the systemic blood pressure occurs. C paths maintain stable perfusion under changing conditions, most probably in an effort to minimally alter the basic metabolic needs of the retinal tissue. Hyperglycemia per se was not responsible for changes of the cell flux in these capillary paths. PMID- 10828735 TI - Concentrations of serum albumin and nonesterified fatty acids in bovine aqueous humor with regard to the fatty acid sensitivity of bovine lens epithelial cells. AB - Concentrations of albumin and nonesterified fatty acids (FAs) have been determined in bovine serum and aqueous humor (AH) with regard to the FA-to albumin molar ratio, which has been proved decisive for the FA cytotoxicity to bovine lens epithelial cells as described previously. In 6- to 7-month-old female calves the average albumin level was reduced 309-fold from 638 micromol/l in serum to 2.00 micromol/l in AH, in 72- to 84-month-old cows 405-fold from 632 to 1. 56 micromol/l, and in 12- to 18-month-old bulls 395-fold from 553 to 1.40 micromol/l. In 64% of the animals the albumin concentration in AH differed significantly between both eyes. The serum albumin values resulted in a Gaussian curve, but the frequency distribution for AH albumin values skewed to the right. Using an enzymatic method the concentrations of nonesterified FAs were determined in serum as well as in concentrated AH samples. In all animals investigated the molar FA-to-albumin ratio was higher in AH than in serum. The results are discussed in view of the assumption that unsaturated nonesterified FAs might be cataractogenic risk factors. PMID- 10828736 TI - Cytokines in aqueous humour and serum before and after corneal transplantation and during rejection. AB - Cytokine profiles in aqueous humour were studied in relation to corneal disease and subsequent corneal graft survival or rejection. Cytokine levels in samples obtained from eyes with clear grafts (n = 59) were all within the normal range. At the time of penetrating keratoplasty (n = 146), intraocular levels of IL-6 were increased in 38% (50/131), most markedly in eyes with previous allograft failure or herpetic stromal keratitis. The level of IL-10 was increased in 1 eye (n = 144) and of IL-4 and IFN-gamma in none. During rejection (n = 10), the levels of IL-6 in aqueous humour were increased in 75% (3/4), of IL-10 in 50% (3/6), of IL-4 in none (0/4) and of IFN-gamma in 40% (2/5). In conclusion, the levels of total protein and IL-6 were increased prior to penetrating keratoplasty in eyes with previous inflammation. These results could however not predict the final outcome of the graft. Increased intraocular levels of IL-6, IL-10 and IFN gamma were observed during rejection. PMID- 10828737 TI - Aqueous dynamics in experimental ab externo trabeculectomy. AB - PURPOSE: This study assessed the decrease in intraocular pressure (IOP), the residual outflow resistance of the trabeculum and the filtration site by histology in enucleated pig and human eyes after ab externo trabeculectomy. METHOD: Measurement of the resistance to aqueous outflow was performed using the constant pressure method before and after ab externo trabeculectomy. RESULTS: The outflow facility was 0.31 +/- 0.13 microl/min/mm Hg in pig eyes and 0.24 +/- 0.08 microl/min/mm Hg in human eyes before surgery, and 79.0 +/- 47. 6 microl/min/mm Hg in pig eyes and 6.33 +/- 6.67 microl/min/mm Hg in human eyes after ab externo trabeculectomy. On histological examinations the aqueous outflow pathway was seen to be at the level of the residual trabecular meshwork. CONCLUSION: This study showed that ab externo trabeculectomy significantly lowers IOP and improves outflow facility in enucleated pig and human eyes. PMID- 10828738 TI - Disjunctive optokinetic nystagmus in a naturally esotropic macaque monkey: interaction between nasotemporal asymmetries of versional eye movement and convergence. AB - OBJECTIVE: To assess the interaction between versional and vergence eye movements in normal and strabismic monkeys. METHODS: Horizontal optokinetic nystagmus (OKN) and vergence were measured using the magnetic scleral search coil technique in a normal adult monkey and a strabismic monkey who had naturally occurring early onset esotropia. Mean eye velocity and vergence angles were calculated during the slow phases of OKN. RESULTS: The strabismic monkey had a nasotemporal asymmetry of OKN favoring nasally directed motion in each eye. During monocular optokinetic stimulation, mean eye velocities were substantially greater for the adducting as compared to the abducting eye. The velocity of the abducting eye was between 55 and 80% of the velocity of the adducting eye (p < 0.01). As a consequence of the disjunctive movements, the eyes converged an average of 4 +/- 2.8 degrees during OKN. Saccadic analysis documented normal lateral rectus function in each eye. Neither an OKN asymmetry nor disjunctive OKN was observed in the normal monkey. CONCLUSION: Disjunctive OKN in the esotropic monkey suggests that the cerebral maldevelopment responsible for nasally biased OKN also contributes to nasal biases in vergence pathways. PMID- 10828739 TI - Evaluation of the norrie disease gene in a family with incontinentia pigmenti. AB - Incontinentia pigmenti (IP) is an ectodermal multisystem disorder which can affect dental, ocular, cardiac and neurologic structures. The ocular changes of IP can have a very similar appearance to the retinal detachment of X-linked familial exudative vitreoretinopathy, which has been shown to be caused by the mutations in the Norrie disease gene. Therefore, it is of interest to determine whether similar mutations in the gene can account for the retinal pathology in patients with IP. To test our hypothesis, we have analyzed the entire Norrie disease gene for a family with IP, by single strand conformational polymorphism followed by DNA sequencing. The sequencing data revealed no disease-specific sequence alterations. These data suggest that ocular findings of IP are perhaps associated with different genes and there is no direct relationship between the genotype and phenotype. PMID- 10828741 TI - 2-Oxo-2H-Pyrimido[2,1-b]benzothiazoles inhibit brain benzodiazepine receptor binding in vitro. AB - 2-Oxo-2H-pyrimido[2,1-b]benzothiazole derivatives were found to inhibit the in vitro binding of (3)H-Ro 15-1788 ((3)H-flumazenil) to rat cortical benzodiazepine receptors with IC(50) values in the range of 0.7-13 micromol/l. The most potent compound, 2-oxo-4-phenyl-2H-pyrimido[2,1-b]- benzothiazole showed a similar potency to inhibit (3)H-Ro 15-1788 binding to membrane preparations of rat brain cortex, cerebellum and hippocampus as well as to various subunit combinations of recombinant human gamma-aminobutyric acid(A)/benzodiazepine receptors. Scatchard plot analysis showed that 2-oxo-4-phenyl-2H-pyrimido[2,1-b]benzothiazole is a competitive inhibitor of (3)H-Ro 15-1788 binding to rat brain cortical membrane preparations. PMID- 10828742 TI - Animal pharmacology of reversible antagonism of the gastric acid pump, compared to standard antisecretory principles. AB - To define the basic antisecretory profile of a potassium-competitive antagonism of the gastric acid pump relative to other classes of acid-inhibitory drugs, they were compared to each other against all three major stimuli of acid secretion. Pumaprazole is an imidazo-pyridine derivative that was used in this investigation as an example of reversibly binding acid pump antagonists (APAs). It differs from covalently binding proton pump inhibitors (PPIs), such as omeprazole, both with respect to chemical structure and mode of interaction with the gastric H(+)/K(+) ATPase (i.e., the acid or proton pump). The present data show that a single dose of pumaprazole is able to elevate intragastric pH in the dog with gastric fistula under pentagastrin or carbachol stimulation from pH 1 to about pH 7 while still displaying a dose-dependent, well-controlable duration of action of a few hours. Ranitidine at the same oral dose also shows a short duration of action, but combined with a far lower efficacy. By contrast, a single oral dose of the PPI omeprazole elevates intragastric pH for a longer time period, but this pH elevation is far lower compared to that of the APA. Regarding the less stringent parameter of inhibition of total acid output in the Heidenhain pouch dog, the modified Shay rat or the Ghosh-Schild rat, pumaprazole is, overall, slightly more efficacious than ranitidine. The M(1)-muscarinic antagonist pirenzepine is ineffective (against histamine stimulation) or far less effective than pumaprazole (against pentagastrin-stimulation), but as effective as pumaprazole against carbachol stimulation in the Ghosh-Schild rat. Basal acid output in the same model is more effectively inhibited by pumaprazole than by ranitidine. In conclusion, our data demonstrate the exceptional ability of a reversibly binding APA to elevate intragastric pH up to neutrality even upon a first administration while still displaying a limited, dose-dependent duration of action. PMID- 10828740 TI - Participation of neuropeptides in antigen-induced contraction of guinea pig bronchi via NK(2) but not NK(1) receptor stimulation. AB - We evaluated the contribution of neuropeptides to antigen-induced contractions of isolated bronchi and tracheae of passively sensitized guinea pigs using CP-96345 and SR 48968, specific antagonists of NK(1) and NK(2) receptors, respectively, in combination with treatment by an antihistaminic and a cysteinyl leukotriene antagonist. SR-48968 but not CP-96345, significantly inhibited the late phase of the bronchial contraction. Phosphoramidon, a neutral endopeptidase inhibitor, tended to potentiate bronchial contraction. Posttreatment with SR-48968 decreased the enhanced contraction induced by the inhibitor as well as the nonenhanced contraction to similar levels of tension. On the other hand, antigen-induced tracheal contraction was not altered by either neuropeptide antagonist. These results suggest that neuropeptides mediate the antigen-induced contractile response of the guinea pig bronchus partly through NK(2) receptor stimulation. PMID- 10828743 TI - Pharmacological analysis of neurogenic, sympathetic responses mediated through alpha-1-, alpha-2-adrenergic and purinergic receptors in the dog saphenous vein. AB - Electrical transmural stimulation evoked a sympathetic contraction in the isolated dog saphenous vein. This contraction consisted of three components (alpha(1)-adrenergic, alpha(2)-adrenergic and purinergic), which were separately observed under combined treatments either with yohimbine (blockade of alpha(2) adrenoceptor) and alpha,beta-methylene ATP (desensitization of P(2X) purinoceptors), with prazosin (blockade of alpha(1)-adrenoceptor) and alpha,beta methylene ATP, or with prazosin and yohimbine, respectively. The alpha(1) adrenergic and purinergic contractions immediately developed after the start of stimulation and reached a peak rapidly. In contrast, the alpha(2)-adrenergic contraction developed slowly, thus the time to peak contraction was longer than the other two components. The relationship between the peak amplitudes of contraction and stimulus frequencies were similar between alpha(1)- and alpha(2) adrenergic components, but the purinergic contraction was smaller than the other components at all frequencies (0.1-30 Hz). Cocaine, a neuronal uptake inhibitor of noradrenaline, significantly potentiated alpha(1)- and alpha(2)-adrenergic components and prolonged their duration with a relatively greater effect on the alpha(2)-adrenergic component. In contrast, the purinergic component was not affected by cocaine. Exogenous noradrenaline produced concentration-dependent contraction, which was inhibited more effectively after combined treatment with combination of prazosin and yohimbine than either blocker given alone. Cocaine potentiated the attenuated contractile response to noradrenaline in the presence of prazosin, resulting in the recovery of response to the control level. Exogenous ATP produced a transient contraction, which was abolished under conditions where postjunctional P(2X)-purinoceptors were desensitized with alpha,beta-methylene ATP. Cocaine did not affect the ATP-induced contraction. These results suggest that sympathetic contraction of the dog saphenous vein is caused through three distinct routes (alpha(1)- and alpha(2)-adrenoceptors and P(2X)-purinoceptors). PMID- 10828744 TI - Structural variants of a human 5-HT1a receptor intracellular loop 3 peptide. AB - To better understand cytoplasmic loop 3/G protein coupling, variations in a bioactive synthetic peptide probe (P1) were constructed according to the published sequences of the human 5HT1a receptor. These probes were tested in a model system of human 5HT1a receptor stably expressed in Chinese hamster ovary cells. In agonist inhibition studies, peptides with amino acid substitutions of residues 6-9 from the amino terminus of loop 3 were less active than P1. Truncated peptide P4, conserving the residue 6-9 region, was also less active than P1. Truncates P5 and P6, deleting the residue 6-9 region, were inactive. When cAMP levels were measured, both substituted peptides were more active than P1 in this negatively coupled system. In contrast, the truncated peptides were without activity in the cAMP assays. Thus, P1 and its derivatives (P2-P6) constitute a small group of peptides with differential uncoupling (agonist inhibition) and signal transduction (cAMP) activities in this G-protein-linked system. It is proposed that these peptides will be useful in future studies detailing the molecular determinants at the receptor/G protein interface. PMID- 10828745 TI - Effects of centrally applied somatostatin on pituitary adrenocorticotropes in female rats. AB - Effects of intracerebroventricular (i.c.v.) application of somatostatin (SRIH-14 or SRIH-28) on growth and function of pituitary adrenocorticotropes (ACTH cells) were examined in adult female Wistar rats. Animals were subjected to i.c.v. administration of three 1-microg doses of SRIH-14 or SRIH-28 dissolved in 5 microl saline every second day. Controls were treated in the same way with the same volume of saline only. ACTH-producing cells were studied using the peroxidase-antiperoxidase (PAP) immunohistochemical procedure; blood samples were collected for hormone analyses 5 days after the last injection. SRIH-28 treatment decreased (p < 0.05) all morphometric parameters compared to control rats. Volume of ACTH cells decreased by 10%, nuclei by 36% and volume density by 13%. No significant changes (p > 0.05) in these parameters occurred after SRIH-14 treatment. Plasma concentration of ACTH in SRIH-28-treated rats was significantly lower (p < 0.05) than in control rats by 35%. In SRIH-14-treated rats, plasma concentration of ACTH was slightly, but not significantly (p > 0.05) increased by 13% compared to saline treatment. These observations suggest that centrally administered somatostatin-28, but not somatostatin-14, is specifically involved in the control of growth and secretory activity of ACTH cells in female rats. Thus, selective pharmacological manipulation of SRIH-28 receptors reached from CSF may affect ACTH activity without altering actions usually attributed to receptors sensitive to SRIF-14. PMID- 10828746 TI - Effect of Am-80, a retinoid derivative, on 2, 4-dinitrofluorobenzene-induced contact dermatitis in mice. AB - Retinoids have many pharmacological activities, including anti-inflammatory action and antiangiogenesis, effected through the regulation of various gene transcriptions. In this study, we investigated the effect of Am-80, one of the retinoic acid derivatives, on hapten-induced contact hypersensitivity in BALB/c mice. After application of 2,4-dinitrofluorobenzene (DNFB) to the ears of the mice, severe contact hypersensitivity with marked infiltration of inflammatory cells and hypertrophy of the epidermis was caused. The thickness of the ears increased biphasically and reached a peak 3 and 24 h after the DNFB challenge. Am 80 significantly inhibited ear thickness in the late-(24 h), but not the early phase (3 h) reaction in a dose-dependent manner. In a histopathological study, obvious depression of edema and infiltration of inflammatory cells was observed in the ears of mice treated with Am-80. Am-80 inhibited the levels of expression in mice ears of interferon-gamma (IFN-gamma) and interleukin-6 (IL-6), but not tumor necrosis factor-alpha (TNF-alpha) or interleukin-4 (IL-4). Furthermore, Am 80 inhibited the antigen-induced production of some cytokines, including IFN gamma and IL-6, but not IL-4, in vitro. Therefore, Am-80 inhibited hapten-induced contact hypersensitivity through the direct inhibition of inflammatory cytokines such as IFN-gamma and IL-6. PMID- 10828747 TI - Chairman's introduction PMID- 10828748 TI - Disease-specific Helicobacter pylori virulence factors: the unfulfilled promise. AB - A number of putative virulence factors for Helicobacter pylori have been identified including cagA, vacA and iceA. The criteria for a true virulence factor includes meeting the tests of biologically plausibility with the associations being both experimentally and epidemiologically consistent. Although disease-specific associations have been hypothesized/claimed, there are now sufficient data to conclusively state that none of these putative virulence factors have disease specificity. CagA has been claimed to be associated with increased mucosal IL-8 and inflammation, increased density of H. pylori in the antrum, duodenal ulcer (DU), gastric cancer, and protection against Barrett's cancer. Only the increase in IL-8/inflammation is direct and substantiated. Different H. pylori strains with functional cag pathogenicity islands do not vary in virulance as it has been shown that mucosal IL-8 levels are proportional to the number of cagA + H. pylori independent of the disease from which the H. pylori were obtained. It is now known that the density of either cagA + and cagA H. pylori in the antrum of patients with H. pylori gastritis is the same. In contrast, the mean density of H. pylori in the antrum in DU is greater than in the antrum of patients with H. pylori gastritis. Of interest, the density of H. pylori is higher in the corpus of patients with H. pylori gastritis than those with DU, suggesting that acid secretion plays a critical role in these phenomena. The presence of a functional cag pathogenicity island increases inflammation and it is likely that any factor that results in an increase in inflammation also increases the risk of a symptomatic outcome. Nevertheless, the presence of a functional cag pathogenicity island has no predictive value for the presence, or the future development of a clinically significant outcome. The hypothesis that iceA has disease specificity has not been confirmed and there is currently no known biological or epidemiological evidence for a role for iceA as a virulence factor in H. pylori-related disease. The claim that vacA genotyping might prove clinically useful, e.g. to predict presentation such as duodenal ulcer, has been proven wrong. Analysis of the worldwide data show that vacA genotype s1 is actually a surrogate for the cag pathogenicity island. There is now evidence to suggest that virulence is a host-dependent factor. The pattern of gastritis has withstood the test of time for its relation to different H. pylori-related diseases (e.g. antral predominant gastritis with duodenal ulcer disease). The primary factors responsible for the different patterns of gastritis in response to an H. pylori infection are environmental (e.g. diet), with the H. pylori strain playing a lesser role. Future studies should work to eliminate potential bias before claiming disease associations. Controls must exclude regional or geographic associations related to the common strain circulation and not to the outcome. The authors must also control for both the presence of the factor and for the disease association. The study should be sufficiently large and employ different diseases and ethnic groups for the results to be robust. The findings in the initial sample (data derived hypothesis) should be tested in a new group (hypothesis testing), preferably from another area, before making claims. Finally, it is important to ask whether the results are actually a surrogate for another marker (e.g. vacA s1 for cagA) masquerading for a new finding. Only the cag pathogenicity island has passed the tests of biological plausibility (increased inflammation) and experimental and epidemiological consistency. PMID- 10828749 TI - Should non-invasive Helicobacter pylori testing replace endoscopy in investigation of dyspepsia? AB - Our knowledge of Helicobacter pylori infection is now changing the way in which we investigate patients presenting with dyspepsia, with noninvasive H. pylori testing replacing endoscopy. Non-invasive H. pylori testing has been shown to be useful in predicting the underlying diagnosis in patients presenting with dyspepsia. Several studies have shown that 20-50% of dyspeptic patients with a positive H. pylori test will have evidence of underlying ulcer disease or duodenitis. In contrast, less than 5% of dyspeptic patients with a negative H. pylori test will have evidence of ulcer disease and in these subjects, the likeliest diagnosis is gastroesophageal reflux disease. This has led to many groups recommending that noninvasive H. pylori testing should be used in place of endoscopy, with all those testing positive being given anti-H. pylori therapy and those testing negative being treated symptomatically. One concern about nonendoscopic management of dyspeptic patients is the possibility of missing underlying malignancy but studies have shown that in western countries this is rare in patients less than 55 years of age presenting with dyspepsia in the absence of sinister symptoms. There is increasing evidence supporting eradication of H. pylori infection in dyspeptic patients without ulcer disease. Meta-analysis of four prospective randomized trials indicates that such treatment is superior to placebo in about 10% of subjects. H. pylori-positive dyspeptic patients are also recognized to have an increased risk of developing ulcer disease in the future which will be removed by treating the infection. Another justification for eradicating the infection in the absence of ulcer disease is the fact that H. pylori infection is now proven to be a risk factor for gastric cancer. Prospective randomized studies comparing endoscopy with noninvasive H. pylori testing in the management of dyspeptic patients indicate that managing dyspepsia by noninvasive H. pylori testing is at least as effective as endoscopic-based management in producing symptomatic resolution and saves a substantial number of endoscopic procedures. There is therefore now substantial evidence indicating that noninvasive H. pylori testing should be used in place of endoscopy to determine the management of younger dyspeptic patients without sinister symptoms and who are not taking nonsteroidal anti-inflammatory drugs. PMID- 10828750 TI - Where are We with current therapy? AB - Despite intensive research and widely publicized recommendations from consensus meetings in different continents, the public and primary care physicians are relatively slow in picking up the impact of Helicobacter pylori infection and identifying optimal therapies. The treatment of H. pylori infection has evolved from bismuth-containing regimens, 2-week proton pump inhibitor (PPI)-dual therapies, and now, the widely accepted PPI/ranitidine bismuth citrate (RBC) single week triple therapies. There is a wealth of evidence showing that these regimens are highly efficacious and well tolerated by patients. The MACH-2 studies have confirmed that the addition of a PPI to two antimicrobials has significantly improved the cure rate of H. pylori infection and reduced the impact of antimicrobial resistance. Attempts to use shorter regimens ranging from 1 to 3 days should be resisted because of their unacceptably low therapeutic efficacy. In the United States, there are some indications that 10-14 days of treatment may be required. While the first-line therapies for H. pylori infection is well established, we are still struggling with the choice of optimal regimen in retreatment after the first attempt fails. Quadruple therapy combining PPI with bismuth, metronidazole and tetracycline has achieved a respectable success of around 85%. Switching between metronidazole and clarithromycin seems to be a sensible strategy as these two are the most effective anti-Helicobacter agents. Changing between PPI and RBC in the triple therapy would not make much difference without replacing some of the antimicrobials. Rifabutin-containing regimens and high-dose PPI-amoxicillin dual therapy deserve more studies with large-scale studies. Data on anti-Helicobacter therapy for children are few. Most studies based on bismuth derivatives in combination with amoxicillin or tinidazole and were limited by the small number of cases. Recent studies showed 1-week bismuth based triple therapy and 2-week PPI-based triple therapy are highly efficacious. Reinfection in children > 5 years of age after successful cure is rare. It is worthwhile to refine the optimal therapy for children as the treatment of this group would, theoretically, prevent the development gastric cancer in the long term. PMID- 10828751 TI - Challenges to therapy in the future. AB - Quadruple therapy (with a proton pump inhibitor (PPI), metronidazole, tetracycline and bismuth) is generally reserved for second-line treatment; however, studies using this regimen for 7 days have found it to be effective even in metronidazole-resistant strains. Resistance is an ongoing problem with antimicrobial therapy but considerable progress has now been made into understanding the underlying genetic mechanisms of this process. Metronidazole resistance in Europe is usually in the range of 20-30% of strains but may be as high as 70% in some countries. One genetic mechanism involved is thought to be a mutation of the rdxA gene. Macrolide resistance appears to be on the increase in Europe, varying from 1% in some countries to 13% in others. The genetic mechanism involved has been shown to be a point mutation of a ribosomal RNA. Amoxicillin resistance is an emerging problem that has an adverse effect on eradication rates in clinical practice. Resistance has been shown to be caused by the absence of one of the four binding proteins in the cell wall. Few novel antibiotics have been developed for use in eradication therapy, although rifabutin, secnidazole and furazolidone have shown some success as part of combination therapy. Alternative therapies that have been tested include mucosal protective agents which have been used in place of a PPI in some eradication regimens with some success, and the somatostatin analog, octreotide, that has been used as part of quadruple therapy in place of a PPI and produced eradication rates of approximately 88%. The ultimate challenge is still to develop a safe and effective vaccine against Helicobacter pylori. Current and future research will also focus on identifying genetic targets for therapy, adhesion molecule analogs to prevent binding of the bacterium, and urease inhibitors. The current triple therapy treatment options available for the eradication of Helicobacter pylori infection are over 90% effective in susceptible organisms and there are very few medical conditions to which we can offer such efficacious treatment. Unfortunately, the recommendations made at consensus conferences are not always put into practice and physicians in primary care may be unaware of the true efficacy of eradication therapy. Treatment is very simple: three drugs, twice a day for 1 week. The main focus for both primary care physicians and gastroenterologists should be to reinforce the need for patient compliance, otherwise we will see an increase in antibiotic resistance. Patients should be prewarned that they may experience some mild side effects and should be encouraged to complete the course of treatment. The real challenge for the future will be the management of patients who do not respond to first-line treatment. This paper will focus on potential problems with therapy, such as antibiotic resistance, and possible future solutions, such as novel antibiotics and vaccines. PMID- 10828752 TI - Panel discussion - unresolved issues PMID- 10828753 TI - Introduction PMID- 10828754 TI - Neurogenic factors and hypertension in renal disease. AB - Hypertension in chronic renal failure (CRF) is very common and contributes to morbidity and mortality and to the progression of renal disease. The pathogenesis of hypertension in CRF has been attributed mostly to sodium retention and to activation of the renin-angiotensin-aldosterone system. More recently an abundance of evidence has accumulated to support a role for increased sympathetic nervous system (SNS) activity in the genesis of hypertension associated with CRF. Evidence from our laboratory has also demonstrated that the rise in central SNS activity is mitigated by increased local expression of nitric oxide synthase (NOS)-mRNA and nitric oxide (NO) production, and that the upregulation of NO production in the brain is mediated by IL-1beta. PMID- 10828755 TI - The role of vasoactive compounds, growth factors and cytokines in the progression of renal disease. AB - A number of kidney diseases, and their progression to end-stage renal disease, are driven, in part, by the effects of angiotensin II. Increasing levels of angiotensin II may in turn up-regulate the expression of growth factors and cytokines, such as transforming growth factor-beta1 (TGF-beta1), tumor necrosis factor-alpha (TNF-alpha), osteopontin, vascular cell adhesion molecule-1 (VCAM 1), nuclear factor-kappaB (NF-kappaB), platelet-derived growth factor (PDGF), basic fibroblast growth factor (bFGF) and insulin-like growth factor. Most of these compounds promote cell growth and fibrosis. Angiotensin II also stimulates oxidative stress. This stress in turn may potentiate the vasoconstrictor effect of the peptide due, in part, to increased catabolism of nitric oxide (NO). Oxidative stress, fueled in part by angiotensin II, up-regulates the expression of adhesion molecules, chemoattractant compounds and cytokines. The angiotensinogen gene, which provides the precursor for angiotensin production, is stimulated by NF-kappaB activation. NF-kappaB is activated by angiotensin in the liver and in the kidney. This provides an autocrine reinforcing loop that up regulates angiotensin production. Angiotensin II activates NF-kappaB through both AT1 and AT2 receptors. In addition, angiotensin-converting enzyme (ACE) inhibition markedly decreases NF-kappaB activation in the setting of renal disease. PMID- 10828756 TI - Glomerular hypertension, abnormal glomerular growth, and progression of renal diseases. AB - The development and progression of sclerosis is determined by complex interactions of many mechanisms, including direct hemodynamic actions, modulation of glomerular cell injury, and growth factor actions. The interplay of these factors determines the balance of cell growth and proliferation versus cell death by necrosis or apoptosis, and the balance of matrix accumulation versus degradation. Sclerosis may even be reversed when therapies inhibit these mechanisms and augment matrix degradation processes, both by directly increasing proteolytic activity and by down-regulating inhibitors of matrix degradation. We will focus in this review on the roles of glomerular hemodynamics and growth in the progression of renal diseases. PMID- 10828757 TI - Is there a common mechanism for the progression of different types of renal diseases other than proteinuria? Towards the unifying theme of chronic hypoxia. AB - The question of why chronic renal diseases progress is a topic only recently investigated. Putative causes such as proteinuria do not account for all aspects of progressive renal disease. An alternative mechanism, chronic hypoxia, is proposed that might better explain certain elements of progressive renal disease, but elements of the hypothesis remain subject to further study. PMID- 10828758 TI - The role of lipids in renal disease: future challenges. AB - Experimental studies have provided in vivo and in vitro data to support the notion that dyslipidemia contributes to glomerular and interstitial injury of the renal parenchyma. The 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors are a new class of lipid-lowering agents that have been extensively studied during the past decade. These agents have significant effects on circulating lipids and both renal and vascular injury. New insights into the mechanisms of action of these agents have revealed an important effect on a variety of inflammatory and fibrogenic processes that appear to have major implications for human renal and cardiovascular diseases. PMID- 10828759 TI - Role of patient factors in therapy resistance to antiproteinuric intervention in nondiabetic and diabetic nephropathy. AB - Reduction of proteinuria is a prerequisite for successful long-term renoprotection. To investigate whether individual patient factors are determinants of antiproteinuric efficacy, we analyzed individual responses to different modes of antiproteinuric intervention in nondiabetic and diabetic patients, obtained in prior studies comparing the efficacy of various pharmacological regimens. The individual antiproteinuric response to angiotensin converting enzyme (ACE) inhibition positively correlated to the response to angiotensin type I (AT1) receptor blockade in diabetic (r = 0.67, P < 0.01, N = 16) as well as nondiabetic patients (r = 0.75, P < 0.01, N = 12). This corresponded to the correlations for antihypertensive efficacy between ACE inhibition and AT1 receptor blockade in diabetic (r = 0.73, P < 0.001) as well as nondiabetic patients (r = 0.55, P < 0.05). Remarkably, the antiproteinuric response to ACE inhibition also correlated positively to the antiproteinuric response to indomethacin (r = 0.63, P < 0.05, N = 9). Thus, patients responding favorably to one class of antiproteinuric drugs also respond favorably to other classes of available drugs, supporting a main role for individual patient factors in responsiveness or resistance to antiproteinuric intervention. In the search for strategies to improve response in these high risk patients, combination treatment (combining different drugs, and combining drugs with dietary measures like sodium and protein restriction), and the use of higher doses may provide more fruitful strategies to optimize renoprotection than shifting to other classes of the available drugs. PMID- 10828760 TI - Dietary therapy in uremia: the impact on nutrition and progressive renal failure. AB - BACKGROUND: In rats with experimental chronic renal failure (CRF), low-protein diets protect against histologic damage and improve mortality. In CRF patients, low-protein diets ameliorate uremic symptoms and certain CRF complications. Fortunately, low-protein diets are nutritionally sound in CRF patients because they activate compensatory mechanisms that conserve protein with a low-protein diet. These results do not determine if dietary protein restriction can slow the rate of progression of CRF or the time to dialysis. METHODS: Reports evaluating low-protein diets and changes in nutritional status and/or progression of CRF are analyzed for efficacy. The MDRD Study is reviewed in depth. RESULTS: When dietary compliance was achieved, the nutritional status was unimpaired and progression was slowed. Studies with limited dietary compliance failed to find any beneficial effect on progression. Problems in study design suggest caution before accepting the initial MDRD Study conclusion that dietary restriction does not slow progression. Subsequent analyses of MDRD results indicate that protein restriction can slow progression of CRF. CONCLUSION: Evidence that dietary protein spontaneously decreases in progressively uremic patients should not be construed as an argument against the use of dietary therapy. Rather, it is a persuasive argument to restrict dietary protein intake in order to minimize CRF complications while preserving nutritional status. In patients with uremia or progression despite other measures, dietary therapy should be started along with monitoring for dietary compliance and nutritional adequacy. PMID- 10828761 TI - The role of systemic hypertension in the progression of nondiabetic renal disease. AB - BACKGROUND: A role for hypertension in the progression of renal disease has been convincingly shown in experimental animals only. In human studies, the relation between hypertension and progression is difficult to demonstrate due to several confounding factors: age, gender, race; the difficult choice of blood pressure (BP) parameters that correlate with progression; the abnormal circadian BP pattern; and the many non-hemodynamic factors of progression. An important role for hypertension in progressive nondiabetic renal disease has been suggested by observational studies and clinical trials originally intended to evaluate the effect of dietary protein restriction on progression. In addition, several studies, summarized by a recent meta-analysis, have shown that pharmacological agents which lower both BP and proteinuria, mainly the angiotensin-converting enzyme inhibitors (ACEI), significantly slow the rate of progression in these diseases. METHODS: In this article we review the effect of lowering BP on the progression of nondiabetic chronic renal disease, the patient characteristics that are associated with a greater or lesser benefit of blood pressure reduction, and the choice of antihypertensive regimens associated with better outcomes in patients with renal disease. RESULTS: Lower levels of achieved BP are associated with a slower decline in renal function, both in patients with and without proteinuria. ACEI are effective BP lowering agents and are associated with better preservation of renal function as opposed to antihypertensive regimens without ACEI. This protective effect of ACEI is in addition to their BP and urine protein lowering effects. The protective effect of ACEI on renal function is more pronounced in patients with proteinuria. CONCLUSION: In patients with nondiabetic renal disease and proteinuria, the risk of progression can be minimized by lowering both BP and proteinuria. ACEI have an additional beneficial effect. PMID- 10828762 TI - The role of underlying nephropathy in the progression of renal disease. AB - BACKGROUND: Disease-specific pathogenic mechanisms may be major determinants of the spontaneous rate of progression of chronic renal failure (CRF). To clarify the role of different underlying renal diseases, we examined the rate of CRF progression in 886 patients with chronic nephropathies. METHODS: Secondary analysis of two multicenter, prospective randomized trials: the Northern Italian Cooperative study (NIC) and the AIPRI study (ACE-Inhibition in Progressive Renal Insufficiency). Univariate and multivariate analyses of variance were used to select the covariates possibly related to CRF progression (estimated by means of the slope of the reciprocal of SCr against time), focusing on the contributory role of primary renal diseases. RESULTS: The overall rate of CRF progression was relatively low but there was a considerable difference in the slopes relating to the underlying nephropathy (particularly evident in the patients with chronic glomerulonephritis (CGN)). The median rate of CRF progression in both studies was more rapid in patients with polycystic kidney disease (PKD) and CGN than in those with other nephropathies. Multivariate analysis showed PKD as an independent predictor of the CRF progression rate only in the NIC Study (P < 0.0015); the selected variables in both studies predicted a variation of only 15-18% in the CRF progression rate. CONCLUSION: The underlying renal disease certainly plays a role in the natural history of CRF, but the variability of the CRF progression rates related to different renal diseases and between individuals with the same diagnosis underlines the need for caution in evaluating risk factors and predicting single patient outcomes. PMID- 10828763 TI - Elevated vascular endothelial growth factor in type 1 diabetic patients with diabetic nephropathy. AB - BACKGROUND: Growth factors have been suggested to play a role in the development and progression of diabetic nephropathy. Vascular endothelial growth factor (VEGF) is a potent cytokine family that induces angiogenesis and markedly increases endothelial permeability. The aim of the present study was to investigate plasma levels of VEGF in a large cohort of type 1 diabetic patients with diabetic nephropathy and in long-standing type 1 diabetic patients with persistent normoalbuminuria, and to evaluate VEGF as a predictor of nephropathy progression. METHODS: We measured VEGF with an enzyme-linked immunosorbent assay (ELISA) technique in 199 type 1 diabetic patients with diabetic nephropathy (122 males, age 41 +/- 10 years, diabetes duration 28 +/- 8 years), glomerular filtration rate (GFR) (median [range]) 75 [10-143] mL/min/1.73 m2, and in 188 long-standing type 1 diabetic patients with persistent normoalbuminuria (115 males, age 43 +/- 10 years, diabetes duration 27 +/- 9 years). One hundred fifty five of the proteinuric patients were followed for at least 3 years after baseline examination with yearly GFR measurements. RESULTS: Plasma levels of VEGF were significantly increased in patients with nephropathy as compared to the normoalbuminuric group; (median [range]): 45.7 [22.0-410] versus 27.1 [22.0-355] ng/L, respectively, P < 0.001. This difference was ascribed to elevated VEGF levels in nephropathic men: 51.8 [22.0-410] versus 22.0 [22.0-308] ng/L, P < 0. 001. No differences were found between women with and without nephropathy: 37.8 [22.0-325] versus 36.6 [22.0-335] ng/L, NS. In proteinuric patients with GFR above and below the median value, there was no difference in the level of VEGF, NS. Plasma VEGF was below the detection limit (22.0 ng/L) in 60 patients with nephropathy and 93 patients with normoalbuminuria, P < 0.001. The mean rate of GFR decline was 3.5 (SE: 0.4) mL/min/year, and the following baseline variables acted as predictors of progression: albuminuria, mean arterial blood pressure and male gender. Hemoglobin A1c and plasma VEGF did not act as predictors. No significant differences between patients with and without proliferative retinopathy were detected. CONCLUSIONS: Our data suggest that VEGF is elevated early in the course of diabetic nephropathy in men with type 1 diabetes mellitus. Baseline albuminuria, arterial blood pressure and male gender was predictors of diabetic nephropathy progression, while plasma VEGF and Hemoglobin A1c did not contribute. The importance of VEGF in the initiation of diabetic nephropathy remains to be established. PMID- 10828764 TI - Progression of renal damage in chronic rejection. AB - A significant complication in renal transplantation is the progressive decline of allograft function, commonly called chronic rejection. This complication has been regarded as a result of continuous immunological activity against the allograft. However, the role of nonimmunological factors has become increasingly apparent in triggering or exacerbating immunological mechanisms. Although the role of immunological factors is predominant, the correction of nonimmunological factors may contribute in improving the probability of long-term graft function. However, immunosuppressive therapy remains the cornerstone for reducing the risk of chronic rejection and late graft failure. PMID- 10828765 TI - Counteracting progression of renal disease: A look into the future. AB - It is the very nature of research, including medical research, that truly novel results are not predictable. Nevertheless, some predictions concerning the understanding and management of progression of renal disease are plausible. It is very likely that in the future, exploding health budgets will force authorities and nephrologists to more effectively apply existing knowledge in this field to patients with early renal disease, particularly diabetics. We hope that this optimistic note is justified, although experience admittedly indicates that it is very much against human nature to behave rationally. With the powerful methodological tools available today, it is safe to predict that insight into the mechanisms underlying progression will also increase. Although pharmacological blockade of the renin-angiotensin system has been one of the great success stories of the past two decades, in many patients, progression is seen despite administration of angiotensin-converting enzyme (ACE) inhibitors and angiotensin receptor blockers. Fortunately, additional targets for intervention, such as endothelin-1, inflammatory cascades, chemokines, etc., are on the horizon. A particularly fertile target for prevention of progression will be kidney grafts, since it has become increasingly clear that factors unrelated to allo-immunity play an important role in chronic allograft nephropathy. PMID- 10828766 TI - Utility of lower extremity venous ultrasound scanning in the diagnosis and exclusion of pulmonary embolism in outpatients. AB - STUDY OBJECTIVE: Emergency physicians frequently rely on normal findings from a lower extremity venous ultrasound examination as a method to decrease the probability of pulmonary embolism (PE) in outpatients with a nondiagnostic ventilation-perfusion lung scan (V/Q scan). The objective of this study was to evaluate the diagnostic utility of bilateral lower extremity venous ultrasound scanning in the diagnosis of PE in emergency department patients with a low-, moderate-, or indeterminate-probability (nondiagnostic) V/Q scan. METHODS: This prospective, 2-center, descriptive study was conducted at the EDs of 2 large teaching hospitals. From an initial cohort of 570 nonreferred outpatients, a convenience sample of 156 patients who had both a nondiagnostic V/Q scan and a lower extremity venous ultrasound scan performed was selected as the study population. The sensitivity and specificity for a single lower extremity venous ultrasound scan and the posttest probability of PE were determined for the study population. RESULTS: In the study population, the best-case sensitivity of the lower extremity venous ultrasound scan for PE was 54% (95% confidence interval [CI] 37% to 71%) and the specificity was 98% (95% CI 94% to 100%). The likelihood ratio of a positive test result was 27. The likelihood ratio of a negative test result was 0.49, yielding a lowest possible posttest probability of PE of 12% (95% CI 6% to 17%). CONCLUSION: This study demonstrates that the combination of a nondiagnostic (low, moderate, or indeterminate) V/Q scan plus a single negative result from lower extremity venous ultrasound examination, even in a best-case scenario, does not exclude the diagnosis of PE. PMID- 10828767 TI - Telephone follow-up after the emergency department visit: experience with acute asthma. On behalf of the MARC Investigators. AB - STUDY OBJECTIVE: This study explored how a variety of demographic and illness related factors were associated with telephone follow-up among patients visiting the emergency department for acute asthma. METHODS: We performed a prospective cohort study as part of the Multicenter Airway Research Collaboration (MARC). The study was performed at 77 EDs in 22 US states and 4 Canadian provinces. ED patients, ages 2 to 54 years, who presented with acute asthma underwent a structured interview during their visit. Two weeks later, research personnel attempted to contact participants by telephone, using numbers obtained during the ED interview. RESULTS: A total of 1,847 adult and 1,184 pediatric patients were interviewed in the ED. Of these, 1,308 (71%) adult patients and 1,026 (87%) pediatric patients were successfully reached for 2-week telephone follow-up. Multivariate analyses revealed the factor most strongly related to contact was age, with pediatric patients being 2.5 times more likely to be reached than adults (95% confidence interval 2.0 to 3.2). Also, participants who were black, low in socioeconomic status, lacking a primary care provider, and exposed to tobacco smoke were significantly less likely to have been reached (all P <.001). CONCLUSION: In contrast to some reports in the literature, telephone contact rates were high. However, successful contact was not equally likely among all patient groups. Although the high contact rates support the feasibility of telephone follow-up of asthmatic patients visiting the ED, the results also act as a reminder of the potential biases that may arise when using telephone contact for clinical, quality assurance, and research reasons. PMID- 10828768 TI - Improving telephone contact rates of patients discharged from the emergency department. AB - STUDY OBJECTIVE: This study investigated the rate of incorrect contact telephone numbers recorded during emergency department registration, and evaluated whether postdischarge contact rates can be improved by verifying the best contact number with the patient before discharge. METHODS: A prospective study was conducted with convenience sampling at a tertiary care hospital with an annual census of 60,000. Patients older than 18 years were enrolled, and the "unverified" telephone numbers recorded at registration were entered on the data sheet. Patients were then asked, "What number can we reach you at to discuss lab or x ray results?" These "verified" numbers and additional demographic data were entered on the data sheet. Within 1 week, 3 calls were made to both the unverified and the verified numbers. Calls were considered successful if the patient, a friend, family member, or coworker was reached, or if the patient returned a message left on an answering machine. RESULTS: Four hundred eighteen patients (43% men) were enrolled; 72 (17%) patients provided a different best contact number than the one recorded on the chart. When unverified numbers were called, only 68.9% of patients were reachable, whereas when verified numbers were called, 81.8% of patients were contacted (P <.01). No statistical difference was found between patients who were successfully contacted and those who were not with regard to age, sex, race, or time of visit. Patients insured by health maintenance organizations were more likely to be reached (P =.02). CONCLUSION: Verification of a best contact telephone number significantly improves the ability to contact patients after ED discharge. PMID- 10828769 TI - Out-of-hospital succinylcholine-assisted endotracheal intubation by paramedics. AB - STUDY OBJECTIVE: To describe the effectiveness and safety of succinylcholine administration by paramedics to out-of-hospital patients requiring endotracheal intubation. METHODS: A consecutive case series was collected by retrospective review of patient care records from a large, private ambulance agency serving an urban and rural county of 600,000 residents over a 40-month period. All patients for whom endotracheal intubation was facilitated by succinylcholine use were identified. Data were systematically collected on demographics, paramedic diagnosis, intubation attempts, and complications. RESULTS: Succinylcholine was used in 150 patients with a mean age (+/-SD) of 50+/-23 years. Paramedic diagnosis was coma in 43% (64/150; 95% confidence interval [CI] 35% to 51%), trauma in 26% (39/150; 95% CI 19% to 33%), and respiratory failure in 31% (47/150; 95% CI 23% to 39%). No patient was in cardiac arrest at the time of succinylcholine use. The intubation success rate was 92% (138/150; 95% CI 88% to 96%), with success achieved on the first attempt in 82% of patients (123/150; 95% CI 76% to 88%). Four patients who received succinylcholine developed cardiac arrest, and 3 others had symptomatic bradycardia. CONCLUSION: The use of succinylcholine by paramedics to assist intubation appears to aid in intubation of patients who are not in cardiac arrest. However, significant cardiac complications can occur. PMID- 10828770 TI - Association between prepayment systems and emergency medical services use among patients with acute chest discomfort syndrome. For the Rapid Early Action for Coronary Treatment (REACT) Study. AB - STUDY OBJECTIVE: Cost concerns may inhibit emergency medical services (EMS) use. Novel tax-based and subscription prepayment programs indemnify patients against the cost of EMS treatment and transport. We determine whether the presence of (or enrollment in) prepayment plans increase EMS use among patients with acute chest discomfort, particularly those residing in low-income areas, those lacking private insurance, or both. METHODS: This study uses a subset of baseline data from the REACT trial, a multicenter, randomized controlled community trial designed, in part, to increase EMS use. The sample includes 860 consecutive noninstitutionalized patients (>30 years old) presenting with nontraumatic chest discomfort to hospital emergency departments in 4 Oregon/Washington communities. The association between prepayment systems and EMS use was analyzed using multivariable logistic regression. RESULTS: Overall EMS use was 52% (n=445). Among EMS users, 338 (75%) were subsequently admitted to the hospital and 110 (25%) were released from the ED. Prepayment was not associated with increased EMS use in the overall patient sample. However, patients residing in low-income census block groups (median annual income <$30,000) were 2.6 times (95% confidence interval [CI] 1.4 to 4.8) more likely to use EMS when a prepayment system was available than when no system was present. No association was noted among higher-income block group residents. Among low-income block group residents lacking private insurance, prepayment systems were associated with 3.8 times (95% CI 1.2 to 13.4) greater EMS usage. CONCLUSION: Economic considerations may affect EMS system utilization among underinsured and low-income patients experiencing a cardiac event. Prepayment systems may increase EMS utilization among these groups. PMID- 10828771 TI - Does transportation by ambulance decrease time to gastrointestinal decontamination after overdose? AB - STUDY OBJECTIVE: Because the ability of gastrointestinal decontamination to alter drug absorption varies inversely with time, we compared the time from arrival in the emergency department to gastrointestinal decontamination (gastric lavage or activated charcoal) for patients transported by ambulance with patients transported by other means after overdose. METHODS: A retrospective chart review was conducted in an academic university ED with an annual volume of 56,000 visits. Consecutive cases of oral overdose treated by gastrointestinal decontamination between December 1, 1995, and May 31, 1996, were identified from International Classification of Diseases, ninth revision, billing codes. ED charts were reviewed to determine the patient's age, sex, mode of transportation, disposition, and time interval to gastrointestinal decontamination with either gastric lavage or activated charcoal. RESULTS: Two hundred eighty-one patient visits were identified. Complete data were available for 173 visits. Six patients were excluded because the diagnosis of overdose was not made on presentation, leaving 167 cases for analysis. The median age for all patients was 27 years; 95 (57%) were female. Overdose patients were transported by ambulance in 105 (67%) cases. Admission rates were similar for patients transported by ambulance and those who arrived by other means. The median interval from arrival to any gastrointestinal decontamination (lavage or charcoal) for patients transported by ambulance was shorter than patients who arrived by other means at 55 and 73 minutes, respectively (95% confidence interval for difference 2.5 to 30.5 minutes, P =.03). Subgroup analysis showed this difference was largely the result of gastric lavage. CONCLUSION: Overdose patients transported by ambulance have a shorter time interval from ED arrival to gastrointestinal decontamination than patients arriving by other means. This difference was largely related to more rapid gastric lavage. PMID- 10828772 TI - Motor vehicle crash characteristics and medical outcomes among older drivers in Utah, 1992-1995. AB - STUDY OBJECTIVE: We sought to compare the characteristics and medical outcomes of motor vehicle crashes for drivers 70 years and older with those of drivers between the ages of 30 and 39 years. METHODS: We probabilistically linked statewide motor vehicle crash and hospital discharge data between the years of 1992 and 1995 for the state of Utah. We calculated the odds of older drivers exhibiting certain motor vehicle crash characteristics compared with younger drivers. Adjusting for nighttime crash, high-speed crash, and seatbelt use, we calculated the odds of an older driver being killed or hospitalized compared with those of a younger driver. RESULTS: During the study years, there were 14,466 drivers older than 69 years and 68,706 drivers between the ages of 30 and 39 years involved in motor vehicle crashes in Utah. Older drivers were less likely to have crashes involving drug or alcohol use (odds ratio [OR] 0.1; 95% confidence interval [CI] 0.1 to 0.2) and less likely to have crashes at high speed (OR 0.6; 95% CI 0.6 to 0.7). Although older drivers were no more likely to have a crash involving a right-hand turn (OR 1.0; 95% CI 0.9 to 1.1) than younger drivers, they were over twice as likely to have a crash involving a left-hand turn (OR 2.3; 95% CI 2.2 to 2.5). Also, older drivers were more likely to be killed or hospitalized than younger drivers (OR, 3.5; P <.001). Among belted drivers, an older driver was nearly 7 times more likely to be killed or hospitalized than a younger driver (OR 6. 9; 95% CI 5.4 to 8.9). CONCLUSION: Older drivers do have distinctive motor vehicle crash patterns. Interventions must be taken to reduce the number of left-hand turn crashes involving older drivers. In addition, further research is needed to design, implement, and evaluate countermeasures that may enable older drivers to continue driving while keeping public safety in the forefront. PMID- 10828773 TI - Acute pain and pain control: state of the art. AB - Recognition and treatment of pain in the emergency department has undergone an evolution in the past decade. Emergency clinicians, educators, and researchers have begun to address the undertreatment of pain as well as challenge the long standing dogmas concerning pain treatment. Well-described barriers, both psychological and educational, contribute to our providing inadequate pain relief. This state-of-the-art update describes the current perception of our practice with regard to pain relief and how it can be modified. Pain and pain control is such a broad and complex topic that only new advances and important principles relevant to the practice of emergency medicine are presented. Headache, pediatric pain, and procedural sedation and analgesia are not covered in this article as they will be addressed in future state-of-the-art articles. PMID- 10828774 TI - Determination of "futility" in emergency medicine. AB - The practice of emergency medicine routinely requires rapid decisionmaking regarding various interventions and therapies. Such decisions should be based on the expected risks and benefits to the patient, family, and society. At times, certain interventions and therapies may be considered "futile," or of low expected likelihood of benefit to the patient. Various interpretations of the term "futility" and its practical application to the practice of emergency medicine are explored, as well as background information and potential application of various legal, ethical, and organizational policies regarding the determination of "futility. "Decisions regarding potential benefit of interventions should be based on scientific evidence, societal consensus, and professional standards, not on individual bias regarding quality of life or other subjective matters. Physicians are under no ethical obligation to provide treatments they judge to have no realistic likelihood of benefit to the patient. Decisions to withhold treatment should be made with careful consideration of scientific evidence of likelihood of medical benefit, other benefits (including intangible benefits), potential risks of the proposed intervention, patient preferences, and family wishes. When certain interventions are withheld, special efforts should be made to maintain effective communication, comfort, support, and counseling for the patient, family, and friends. PMID- 10828775 TI - Linking data for injury control research. PMID- 10828777 TI - Viral myocarditis presenting with seizure and electrocardiographic findings of acute myocardial infarction in a 14-month-old child. AB - Acute viral myocarditis is an uncommon but potentially fatal illness in children. Patients with myocarditis may present with nonspecific symptoms or atypical findings that make diagnosis in the emergency department difficult. We describe a previously healthy 14-month-old child with difficulty breathing and a tonic clonic seizure who was subsequently found to have ECG changes and cardiac marker elevation consistent with acute myocardial infarction. The patient was immediately transferred from our community hospital ED to our tertiary care children's hospital. Shortly after admission, the patient developed intractable nonperfusing ventricular arrhythmias necessitating extracorporeal membrane oxygenation. Cardiac function did not recover, and the patient required heart transplantation before cessation of bypass. Serology and anatomic pathology confirmed coxsackievirus B myocarditis. This case illustrates (1) the nonspecific presentation of myocarditis as dyspnea and seizure, (2) the manner in which myocarditis can mimic myocardial infarction, and (3) the importance of early diagnosis in the ED and transfer to a tertiary care facility. PMID- 10828776 TI - Futility-futilis-the leaky vessel. PMID- 10828778 TI - National Highway Traffic Safety Administration (NHTSA) notes: EMS education agenda for the future: a vision for the future of EMS education. PMID- 10828779 TI - Letter from the lady on bed 3. PMID- 10828780 TI - Use of the National Institutes of Health Stroke Scale in the emergency department setting. PMID- 10828781 TI - Management of calf deep venous thrombosis. PMID- 10828782 TI - Carbon monoxide poisoning: how about more precise analytic and preanalytic techniques? PMID- 10828783 TI - Carbon monoxide poisoning: how about more precise analytic and preanalytic techniques? PMID- 10828784 TI - Good samaritan protection. PMID- 10828785 TI - Public training in cardiopulmonary resuscitation and public access defibrillation. PMID- 10828786 TI - Commentary PMID- 10828787 TI - Mechanical and dynamic aspects of voice production as related to voice therapy and phonosurgery. AB - Laryngeal framework surgery can change the position and tension of the vocal folds safely without direct surgical intervention in the vocal fold proper. Some 23 years of experience with phonosurgery have proved its usefulness in treating dysphonia related to unilateral vocal fold paralysis, vocal fold atrophy, and pitch-related dysphonias. Meanwhile, much information about the mechanism of voice production has been obtained through intraoperative findings of voice and fiberscopic examination of the larynx. Based on such knowledge together with information obtained through model experiments, the human vocal organ was reconsidered mainly from the mechanical view point, and the roles of voice therapy and singing pedagogy were discussed in relation to phonosurgery. The vocal organ may not be an ideal musical organ and is rather vulnerable, but its potential is enormous. PMID- 10828788 TI - Plastipore implants in the surgical treatment of atrophic rhinitis: technique and results. AB - BACKGROUND: Atrophic rhinitis is a distressing and socially crippling disease. Surgical treatment has been used for patients with severe disease that is unresponsive to medical therapy. METHODS: Eight patients with primary atrophic rhinitis were treated surgically by narrowing of the nasal fossae with implantation of 2 Plastipore plates per affected side into submucosal pockets into the floor of the nose and septum, thus reducing the volume of the nasal fossae. RESULTS: Excellent results were obtained in 6 patients, with complete resolution of symptoms and good results and only minor crusting in 2 patients. One implant was extruded spontaneously 18 months after implantation and was reimplanted. Results in this patient before and after reimplantation were excellent. There were no cases of infection and no other complications. CONCLUSION: We implanted Plastipore, a high-density polyethylene sponge with micropores, which enables tissue ingrowth, anchoring the implants to the surrounding tissue. The surgical technique is performed with the patient under local anesthesia and is relatively easy and avoids complicated flap procedures, allogeneic bone harvesting, and the discomfort of nostril closure for prolonged periods. PMID- 10828789 TI - Intraoral removal of the submandibular gland: a new surgical approach. AB - The transcervical procedure used for treating sialolithiasis in the submandibular gland has been generally accepted by head and neck surgeons. However, several clinical problems after surgery through the transcervical approach have been described. We introduce a new surgical approach for excision of the submandibular gland indicated in the chronically inflamed salivary gland with or without calculus and benign mixed tumor of the submandibular gland. Thirty-one cases of submandibular gland excision through the intraoral approach were reviewed, analyzing surgical technique and morbidity. Early postoperative discomforts developed, such as a temporary lack of function of lingual nerve and a temporary limitation of tongue movement, but recovery was within a short period of time in all patients involved. No symptomatic late complications appeared, such as residual inflammation of Wharton's duct and neurologic sequelae. We suggest that this approach can be extended to the excision of the submandibular gland as an alternative to the transcervical approach. The major advantages of this approach are the avoidance of an external scar and injury to the marginal mandibular nerve or the hypoglossal nerve. PMID- 10828790 TI - Laser soldering and welding for ossicular reconstruction: an in vitro test. AB - To evaluate the possibility of laser application in ossicular reconstruction, soldering and welding of bony tissue with proteineous materials was performed by using a CO (2) laser, and the bonding strength of these materials was measured with a gradual weighing system. The soldered and welded surface of bone was observed with a scanning electron microscope. Additionally, welding of human ossicles and synthetic materials was experimentally tried with lasers, and morphologic changes were observed and recorded. A scanning electron microscopic examination was also done. A 40% albumin solution and commercial fibrin glue showed the best bonding strength. Bridging of denatured protein solder may be a mechanism of bonding. Polycel was melted by using a laser beam, and melted Polycel covered the surface of the ossicle. However, the bonding strength was not enough to use, and exact measurement of bonding strength was not possible. These results suggested the possibility of laser soldering and welding in ossicular reconstruction, especially with ossicle or cortical bone. PMID- 10828791 TI - Frameless computer-aided surgery system for revision endoscopic sinus surgery. AB - To increase the intraoperative safety factor and to acquire anatomic assistance during revision endoscopic sinus surgery (RESS), we used an optical computer aided surgery (CAS) system that we developed collaboratively in Bern, Switzerland. During 1 year, 25 RESSs were performed with CAS: recurrent polyposis (n = 20), recurrent frontal recess stenosis (n = 3), and recurrent frontal recess stenosis with mucocele (n = 2). These patients were compared with a control group of 10 patients undergoing RESS without CAS. The same surgeon (M.C.) performed all operations, and there were no minor or major complications in either group. The clinical inaccuracy of our system is between 0.5 and 2 mm with paired-point and surface matching. The navigation system is an important aid to surgeons in identifying anatomic landmarks that are typically difficult to visualize in this type of surgery, thus reducing the stress placed on the surgeon. PMID- 10828792 TI - Evaluation of the caloric test by combining 3 response parameters. AB - The slow-phase velocity of nystagmus is one of the most sensitive parameters of vestibular function and is currently the standard for evaluating the caloric test. However, the assessment of this parameter requires recording the response by using nystagmography. The aim of this study was to evaluate whether frequency and duration of the caloric nystagmus, as measured by using a clinical test with Frenzel glasses, could predict the result of the recorded test. The retrospective analysis of 222 caloric test results recorded by means of electronystagmography has shown a good association between the 3 parameters for unilateral weakness. The asymmetry observed in the velocity can be predicted by a combination of frequency and duration. On the other hand, no relationship was observed between the parameters for directional preponderance. These results indicate that a clinical caloric test with frequency and duration as parameters can be used to predict the unilateral weakness, which would be obtained by use of nystagmography. We propose an evaluation of the caloric test on the basis of diagrams combining the 3 response parameters. PMID- 10828793 TI - Frey syndrome treatment with botulinum toxin. AB - The goal of this work is to present our results of the intradermic infiltration with botulinum toxin in patients with Frey syndrome. Sixteen hemifaces in 15 patients were studied. Gustatory stimulation was evoked by sucking on a slice of lemon while measurements were done on both hemifaces, with the normal side being used as a control. Skin temperature and color (erythema) were measured with a digital surface thermometer and a skin chromameter, respectively. Sweat quantity and surface were measured by using the previously described blotting paper and iodine-sublimated paper histogram methods, respectively. Testing was repeated 2 weeks after skin infiltration with botulinum toxin (dilution of 50 U/mL). The interinjection distances were 1 cm, and 0.1 mL (5 U) was infiltrated at each injection site. Frey syndrome complaints disappeared in all patients. Small residual amounts of sweat were measurable. The difference in sweat quantity before and after botulinum toxin infiltration was significant in every patient (P < 0.001). Skin temperature and color measurement gave inconclusive results. In conclusion, Frey syndrome treatment with botulinum toxin is an efficient and well tolerated technique. Further work should address the optimal injection parameters. PMID- 10828794 TI - Extended high-frequency ototoxicity induced by the first administration of cisplatin. AB - The ototoxicity induced by cisplatin results in high-frequency hearing loss. The sound-pressure thresholds at extended high frequencies of 8 to 20 kHz were measured in 12 patients with head and neck cancer before and after the first administration of cisplatin. Ototoxicity was defined statistically by the newly introduced regression-line analysis. This analysis revealed that the threshold of 1 of the 12 patients increased evenly from 8 to 10 kHz, and the damage to the hearing system of this patient was permanent. The reasons for such a low rate of ototoxicity might be the long-term administration of low doses of cisplatin and the addition of fosfomycin. PMID- 10828795 TI - Correlation of galectin-3/galectin-3-binding sites with low differentiation status in head and neck squamous cell carcinomas. AB - The accurate determination of levels of differentiation is of prognostic value in human head and neck squamous cell carcinomas (HNSCCs). Because the deliberate selection of biochemical determinants accompanying certain stages of differentiation can refine the predictive power of histochemical assessments, the application of the quantitative evaluation of staining distribution and intensity by computer-assisted microscopy is one prerequisite to potential improvements. We used 2 innovative approaches with peanut agglutinin based on encouraging results with respect to common lectin-histochemistry. First, we used a custom-made neoglycoprotein to monitor the presence of Thomsen-Friedenreich (T) antigen binding sites. Second, we measured the presence of 2 galectins immunohistochemically and, at the same time, measured lectin-histochemically the presence of accessible ligands for the endogenous lectins. We also monitored the presence of calcyclin, a protein with relevance to cell cycle progression or exocytosis. With 61 cases of HNSCC as their basis, including 31 oral, 20 laryngeal, and 10 hypopharyngeal lesions, the data show that the main modifications observed in connection with a loss of differentiation are related to a modification in the levels of both galectin-3/galectin-3-binding site and T antigen/T-antigen-binding site expressions. The data obtained also suggest that galectin-3 could act as an acceptor site for the T antigen. Because the level of differentiation is known to be indicative of the recurrence rate in HNSCCs and our data clearly indicate that galectin-3 and the T antigen (and their respective binding sites) are involved in dedifferentiation processes, further investigation is warranted into the roles of galectins in HNSCC tumor progression and recurrence analysis. PMID- 10828796 TI - Characteristics of sinus-related pain. AB - OBJECTIVES: The goal of this study was to determine possible distinctive features of facial pain when caused by chronic sinusitis and to validate the pain characteristics previously described in the literature. METHODS: Included were 82 patients with radiographic and endoscopic evidence of chronic sinusitis and significant facial pain who underwent functional endoscopic sinus surgery and were available for 1-year follow-up. A modified McGill pain questionnaire was filled out before surgery, and follow-up data were obtained at 1 year. RESULTS: At 1 year 38% of patients had persistent facial pain despite the lack of any evidence of persistent sinusitis. A consistent use of pain adjectives and other distinctive features was noted in patients reporting improvement of headache. There was no correlation between the severity of pain and the extent or location of mucosal disease. The site of pain did not correlate with the site of disease. CONCLUSIONS: Sinusitis-related pain has distinctive features that set it apart from primary headache disorders and other causes of facial pain. Nonsinus causes account for the headache in 1 of 3 patients undergoing sinus surgery. PMID- 10828797 TI - Multinodular goiter: the surgical procedure of choice. AB - Surgical management of multinodular goiter (MNG) poses an ongoing dilemma between radical resection with its associated complications and partial resection, which carries the risk of recurrence and increased morbidity and difficulty for rethyroidectomy. This study was designed to evaluate the recurrence rate and need for reoperation in a carefully selected population of MNG patients, after nontotal thyroidectomy. The study addressed a highly selected population of patients who were treated and thoroughly evaluated at one surgical department for several years. We analyzed the recurrence rate of MNG in 124 patients. The follow up duration extended from 6 to 516 months (mean 93 months). The general recurrence rate for all nontotal bilateral thyroidectomies was 21% (21/100 patients), increasing from 13.4% to 60% according to the extent of resection. The average time for recurrence was 105 months (8.75 years). Among the patients with recurrent MNG, only 4 (4% of the patients with nontotal bilateral thyroidectomy) required secondary surgical interventions with no resultant morbidity. In our series of very highly selected patients, the recurrence rate for nontotal thyroidectomy was high (21%); however, the need for secondary surgical intervention was low (4%). Thus nontotal thyroidectomy for MNG is legitimate. However, we suggest that the surgical procedure of choice be tailored according to the severity of the disease and the patient's general condition. PMID- 10828798 TI - Relation of biochemical, cytologic, and morphologic parameters to the result of gammagraphy with technetium 99m sestamibi in primary hyperparathyroidism. AB - The aim of this study was to analyze the possible relation of biochemical, cytologic, and morphologic parameters to the results of parathyroid gammagraphy with Tc 99m sestamibi in primary hyperparathyroidism. We studied 46 consecutive patients with primary hyperparathyroidism who were undergoing surgery. All the patients were given a preoperative parathyroid gammagraphy with Tc 99m sestamibi and a complete preoperative biochemical study. During the surgical intervention we recorded the weight and size of the pathologic glands to calculate the volume of each. We also determined the percentage of the chief and oxyphil cells in the pathologic glands. Tc 99m sestamibi sensitivity is higher in adenomas (91%) than in hyperplasia (67%) or double adenomas (50%). No relation was found between biochemical or cytologic parameters and gammagraphic results. Weight and gland volume were significantly greater for adenomas than for hyperplasia (P < 0.0014 and P < 0.0004, respectively), and statistically significant differences in both of them were observed between the glands with positive and negative sestamibi. PMID- 10828799 TI - Oxygenation of advanced head and neck cancer: prognostic marker for the response to primary radiochemotherapy. AB - Recent clinical studies suggest that the degree of tumor oxygenation may be predictive of the response of radiation therapy for cancer. In an exploratory investigation of cervical lymph node metastases in 27 patients with advanced squamous cell carcinoma of the oropharynx and hypopharynx, this relationship was investigated by means of oxygen measurements with an Eppendorf PO (2) histograph. The measurements were made before the start of radiation therapy and after the first week of therapy. Clinical response was evaluated 6 weeks after the completion of therapy. Before therapy, marked hypoxia was observed in the lymph node metastases, with a mean PO (2) value of 16.1 +/- 8.2 mm Hg and a hypoxic fraction (PO (2) < 10 mm Hg) of 56.4% +/- 20.0%. After the first week of radiation (9 Gy) there was a general reoxygenation (DeltaPO (2) = 5.0 +/- 10.1 mm Hg, P < 0.05; Deltahypoxic fraction = -11.3% +/- 31.3%, P = 0.11). A relationship between the degree of reoxygenation and tumor response was not observed. Patients without at least partial lymph node response (n = 8) showed poorer pretherapeutic oxygenation (PO (2) mean = 11.1 +/- 2.9 mm Hg) than those who responded to the therapy (n = 19, PO (2) mean = 18.2 +/- 8.8 mm Hg). In this investigation of a defined set of patients with advanced carcinoma of the oropharynx and hypopharynx, we found that pretherapeutic oxygenation data are predictive for the therapeutic response to radiation therapy or radiochemotherapy. PMID- 10828800 TI - Reconstruction of oral defects with the pedicled buccal fat pad flap. AB - Pedicled buccal fat pad flaps were used to reconstruct defects produced by resection of tumors of the oral cavity in 21 patients. The indications included defects of the oral mucosa and defects after excision of a benign or malignant tumor, preferably smaller than 5 cm in diameter. The results were excellent, and there was no added morbidity. Only 1 delay failure and 1 complication were observed. The anatomy and surgical technique are described. The results suggest that these flaps are a direct, convenient, and reliable option for the reconstruction of small defects of the oral mucosa. PMID- 10828801 TI - Prognostic value of p53, proliferating cell nuclear antigen, and Ki-67 expression in undifferentiated nasopharyngeal carcinomas. AB - In this study the prognostic importance of p53, proliferating cell nuclear antigen (PCNA), and Ki-67 expression was analyzed along with the clinical parameters in 35 consecutive patients with undifferentiated nasopharyngeal carcinomas. Immunohistochemistry was used to detect p53, PCNA, and Ki-67 staining. Among the clinical findings, stage IV disease (P = 0.01), cranial nerve paralysis (P = 0.02), and lymph node metastasis (P = 0.06) were associated with shorter survival. The p53 positivity correlated with the presence of lymph nodes, but it was not a significant factor to predict the outcome. PCNA expression was not found to be a prognostic indicator. On the other hand, the proliferative value of Ki-67 staining was suggestive of prognosis. A proliferation index of Ki 67 less than 10% indicated longer survival (P = 0.03). There was no correlation between Ki-67 staining and PCNA index. As a result, the prognostic value of Ki-67 may alert the physician to more aggressive and adjuvant treatment modalities. PMID- 10828802 TI - Endoscopic management of cholesteatoma: long-term results. AB - OBJECTIVES: This report evaluates long-term results of transcanal endoscopic management and surveillance of cholesteatoma. METHODS: Sixty-nine ears with acquired cholesteatoma underwent endoscopic transcanal tympanotomy and atticotomy to access and completely remove the sac. Reconstruction with a composite tragal graft was performed in 38 ears, and the cavities were packed open in 31 ears. Office-based endoscopic surveillance and follow-up were performed. RESULTS: Forty three ears were operated on with the patient under local anesthesia, and 58 were done on an outpatient basis. Three cases were converted into postauricular tympanomastoidectomy. There were no iatrogenic facial nerve injuries. Bone thresholds were stable, except in 1 patient with perilymphatic fistula. Mean follow-up was 41 months, and 19 ears underwent 5 years of follow-up. Six ears required revision surgery, and 9 required office-based minor procedures. CONCLUSIONS: An endoscopic technique allows transcanal, minimally invasive management and surveillance of cholesteatoma with long-term results that compare well to those of postauricular methods. PMID- 10828803 TI - Malignancy of the temporal bone and external auditory canal. AB - A retrospective study of 18 patients with malignant tumors of the external auditory canal and temporal bone was undertaken to gain an Asian perspective of this rare disease. Of these patients, 15 (83%) had squamous cell carcinoma (SCC) and 61% had stage T3 tumors at presentation. The mean age was 56 years (range 38 82 years). Seven (39%) of the 18 patients had radiation-associated tumors (RATs), and all had undergone radiotherapy for treatment of nasopharyngeal carcinoma. The 1-year cumulative recurrence for the RAT group was 100%, but there was no recurrence in the non-RAT group (P = 0.001). In malignancies of the external auditory canal and temporal bone, a different classification and staging system for patients with RATs may be warranted to better guide treatment strategies. PMID- 10828804 TI - Percutaneous bone-anchored hearing aids at a pediatric institution. AB - OBJECTIVES: The goals were to evaluate hearing, complications, and patient satisfaction with the percutaneous bone-anchored hearing aid (BAHA) and to monitor long-term successful use achieved by careful patient selection. METHODS: This prospective longitudinal study included participants with inoperable congenital bilateral aural atresia, pure-tone average (PTA) bone conduction less than 45 dB HL, prior use of a conventional bone-conduction hearing aid (CBCA), and adequate intelligence, integration, and personal hygiene. Surgery for BAHA implantation was performed in two stages. Evaluation consisted of skin reactions, audiologic results with CBCA and BAHA, and patient satisfaction. Follow-up was at least 24 months. RESULTS: In 11 participants aged 5 to 17 years, the PTA free field air conduction improved 37%, and free-field speech discrimination improved 23%. Successful integration and implant use were achieved in 10 cases but were lost in 1 case as a result of head trauma. All patients preferred the BAHA as opposed to the CBCA. CONCLUSIONS: The BAHA is a valuable device that can improve hearing and provide significant parent and patient satisfaction. Careful selection appears to correlate with successful long-term use. PMID- 10828805 TI - Complications after total laryngectomy in nonradiated laryngeal and hypopharyngeal carcinomas. AB - To study the complications of total laryngectomy, we evaluated 471 previously untreated patients who underwent total laryngectomy between 1980 and 1997. This series consisted of 358 patients with primary carcinoma of the larynx and 113 with carcinoma of the hypopharynx. Concurrent neck dissection was performed in 85% of patients. Complications were studied in relation to age, T and N stage, previous tracheostomy, neck dissection, margins, reconstruction, tracheoesophageal puncture, and surgeon. Complication treatment and hospitalization were also evaluated. The overall complication rate was 30.7%, with 29.2% major and 6.5% minor complications. The mortality rate was 0.6% (3/471). Pharyngocutaneous fistula was the most frequent wound complication (21%), followed by wound infection (4.2%) and hemorrhage (2.3%). Pneumonia (1.4%) and embolism (0.4%) were the most frequent medical complications. Hypopharyngeal tumors, neck dissection, and extended procedures had a significantly higher rate of complications. Complication causes, prevention, and treatment are discussed. PMID- 10828806 TI - Evaluation of time of nasal packing after nasal surgery: a randomized trial. AB - A randomized, prospective trial was carried out to compare the rate of hemorrhagic complications after removal of nasal packing left for 24 and 48 hours. A total of 104 patients underwent nasal surgery and were randomly assigned to 1 of 2 groups: group 1, 24 hours of nasal packing; group 2, 48 hours of nasal packing. No statistical difference was observed between the groups in terms of occurrence of hemorrhagic complications. Hypertension was the only prognostic factor for postoperative bleeding. The routine use of 48-hour postoperative nasal packing after nasal surgery is not justified because of the low incidence of bleeding and the potential associated morbidity. At the end of the procedure, surgeons should evaluate the risk of postoperative bleeding (ie, presence of hypertension) and decide whether 24-hour nasal packing is enough. PMID- 10828807 TI - Epithelial differentiation in developing murine eustachian tube and middle ear. AB - Detailed information on how an epithelial differentiation occurred in the developing eustachian tube and middle ear would be helpful in understanding both normal physiology and pathology of the tubotympanum. This study was undertaken to establish patterns of laminin and E-cadherin in the embryonic mouse eustachian tube and middle ear by use of immunohistochemistry at a stage when epithelial differentiation is taking place. This study was also designed to clarify the role of the middle ear mesenchyme. During the development of the eustachian tube, relatively high immunoreactivity to laminin was observed in the epithelium at gestational days 16 and 17, when the developments of ciliated and secretory cells were first observed. At the time of birth, in contrast to epithelium of the eustachian tube, epithelium of the middle ear cavity showed predominant expression of laminin and E-cadherin. These findings suggest that the expressions of laminin and E-cadherin may be correlated with maturation of the epithelium in the eustachian tube and middle ear and that the epithelial differentiation of the developing murine eustachian tube and middle ear may be controlled by epithelial mesenchymal interaction and cell-to-cell interaction. PMID- 10828808 TI - Surgery for secondary hyperparathyroidism in patients undergoing dialysis. AB - Despite recent advances in the diagnosis and therapy of patients with chronic renal failure and secondary hyperparathyroidism (HPT), 5% of these patients may need parathyroidectomy. The purpose of this article is to present our experience with parathyroid surgery in 30 patients with chronic renal failure at "La Paz" University Hospital, analyzing the clinical and biochemical evolution after surgery as well as the recurrence rate. In the first month after surgery, calcium, parathyroid hormone, phosphorus, and alkaline phosphatase levels, as well as bone pain and pruritus, all decreased significantly. Within the first postoperative year, 24 patients remained asymptomatic, and no recurrent secondary HPT was detected. Within the second year after surgery, 15 patients were asymptomatic, and 3 patients showed a recurrence. According to these results, parathyroidectomy is an appropriate surgical procedure for patients with severe overt secondary HPT. PMID- 10828809 TI - Cryopreserved, irradiated tracheal homograft transplantation for laryngotracheal reconstruction in human beings. AB - Subglottic tracheal stenosis is a common clinical entity. Management in severe cases is often problematic. Various techniques for tracheal replacement have been used with varying degrees of success. In this study we used cryopreserved, irradiated tracheal homografts, the use of which in human beings has not been reported previously. In a sterile setup, the tracheas were harvested from donor cadavers within 24 hours of death. The grafts were initially kept at 57 degrees C for 20 minutes; they were then placed in a -70 degrees C chamber for another 2 to 3 days or more and were irradiated to 25 kGy (2.5 million rad). Finally, the grafts were stored at -70 degrees C until usage. Seven patients underwent the surgery, but only 4 are presented here. In the remaining 3 patients, the follow up time was too short to be evaluated. Four patients, 2 male and 2 female (aged 2 40 years, mean 16 years), with severe subglottic tracheal stenosis underwent segmental tracheal graft reconstruction. Immunosuppressant medications were not given to any patient. Follow-up ranged from 18 to 20 months. Three patients successfully underwent decannulation, and 1 patient had local infection and dislodgment of the intraluminal stent with subsequent restenosis. The postoperative tracheal lumen appeared to be near normal, with histologic evidence of normal respiratory epithelium at the grafted site. In conclusion, cryopreserved, irradiated tracheal homograft transplantation is a valuable alternative for subglottic tracheal reconstruction. PMID- 10828810 TI - Accuracy of fine-needle aspiration biopsy of the thyroid combined with an evaluation of clinical and radiologic factors. AB - Fine-needle aspiration biopsy (FNAB) of thyroid nodules is an operator interpreter-dependent procedure whose diagnostic accuracy has deficiencies, especially concerning indeterminate and negative results. There are reports in the literature of up to a 15% definitive diagnosis of carcinoma in the first category of cytology and an 11% false-negative rate in the second category. With these facts in mind, a prospective study was done at the Instituto Nacional de Cancerologia, Mexico City, to determine whether one or several clinicoradiologic factors were associated with a final pathologic diagnosis of carcinoma, thereby increasing the diagnostic accuracy of thyroid FNAB. In total, 200 cases were reviewed, and 159 patients underwent surgery. Our study group comprises these 159 patients. The only clinical factor associated with false-negative results was size > or = 4 cm, with no statistical significance for the other factors analyzed. A comparison was done between clinical factors of patients who had indeterminate results in the aspirate and those of patients who had a definitive diagnosis by permanent section of the thyroid carcinoma, where an association was found between malignancy and tumor size > or = 4 cm, irregular borders, fixed lesions, and heterogeneity on ultrasound. We conclude that in the presence of a negative cytology result, with tumor size > or = 4 cm, close follow-up is indicated, and a new thyroid FNAB should be performed in 6 months. If the nodule persists or grows, a surgical procedure should be considered. In the case of an indeterminate cytology result and in the absence of those factors associated more frequently with a pathologic result of carcinoma, a conservative approach could be indicated with clinical follow-up and repeat thyroid FNAB in 6 months to 1 year. PMID- 10828811 TI - Metastatic carcinoma to the jugular foramen. PMID- 10828812 TI - False aneurysm after rhinoplasty: an unusual complication. PMID- 10828813 TI - Rhinolalia as a symptom of pneumomediastinum after radical neck dissection. PMID- 10828814 TI - Plexiform neurofibroma of the sublingual gland. PMID- 10828815 TI - Practical approach to foreign bodies in the frontal sinus. PMID- 10828816 TI - Desmoid tumor of the neck. PMID- 10828817 TI - Hepatocarcinoma: metastasis to the nose and paranasal sinuses. PMID- 10828818 TI - Use of ototopical antibiotics in treating 3 common ear diseases. AB - Prompted by rising rates of antibiotic resistance, lack of standardized treatment regimens, and new treatment alternatives, the American Academy of Otolaryngology Head and Neck Surgery convened an expert consensus panel to consider recommendations for the responsible use of antibiotics in chronic suppurative otitis media, tympanostomy tube otorrhea, and otitis externa. The Panel concluded that in the absence of systemic infection or serious underlying disease, topical antibiotics alone constitute first-line treatment for most patients with these conditions, finding no evidence that systemic antibiotics alone or in combination with topical preparations improve treatment outcomes compared with topical antibiotics alone. Topical preparations should be selected on the basis of expected bacteriology and informed knowledge of the risk-benefit of each available preparation. The use of nonototoxic preparations in treating acute otitis externa (when the tympanic membrane is perforated or its status is unknown), chronic suppurative otitis media, and tympanostomy tube otorrhea should be considered. PMID- 10828819 TI - Tonsillar lipoma. PMID- 10828820 TI - Use of the endostitch device for closure of pharyngeal traumatic lacerations. PMID- 10828821 TI - Growth and development of the islets of langerhans: implications for the treatment of diabetes. A special section consisting of proceedings from the journal of molecular endocrinology symposium at the 190th meeting of the society for endocrinology, london, 8 november 1999 PMID- 10828822 TI - Islet growth and development in the adult. PMID- 10828823 TI - IGF-I and GH post-receptor signaling mechanisms for pancreatic beta-cell replication. AB - Certain nutrients, pharmacological agents and growth factors can stimulate pancreatic beta-cell proliferation; however, mitogenic signal transduction pathways in beta-cells have not been particularly well characterized. As a model system we have focussed on characterizing the signal transduction pathways immediately downstream of the IGF-I and GH receptors in beta-cells. The original idea was to gain an idea of important elements in mitogenic signaling pathways which might then be exploited to generate a marked increase in beta-cell proliferation. Such an approach could eventually reveal a means to increase the number of human pancreatic endocrine cells in vitro, in order to obtain an abundant source of beta-cells for routine transplantation therapy of type-I diabetes. However, in the course of our studies, we have also unveiled an unexpected insight into the pathogenesis of obesity-linked type-II diabetes. It has been observed that free fatty acids inhibit glucose- and glucose-dependent IGF-I/GH-induced beta-cell proliferation. We hypothesize that a gradual accumulation of intracellular fat in beta-cells during obesity can eventually lead to an inhibition of beta-cell mass expansion and hence failure to compensate for peripheral insulin resistance, so that type-II diabetes ensues. PMID- 10828824 TI - Persistent hyperinsulinaemic hyperglycaemia of infancy-derived cells; implications for beta-cells that replicate in vitro. PMID- 10828825 TI - Expression of 11beta-hydroxylase and aldosterone synthase genes in the rat brain. AB - The terminal stages of cortisol and aldosterone production in the human adrenal gland are catalysed by the enzymes 11beta-hydroxylase and aldosterone synthase, which are encoded by the CYP11B1 and CYP11B2 genes respectively. Recent studies have suggested that aldosterone and cortisol are also made in other tissues such as the brain, heart and vascular system and may play a role in cardiovascular homeostasis. The aim of this study was to confirm the presence of these enzymes and localise them precisely in the rat brain. Reverse transcription-polymerase chain reaction (RT-PCR)/Southern blotting confirmed transcription of CYP11B1 and CYP11B2 in whole brain and hypothalamus minces from Wistar-Kyoto rats. 11beta Hydroxylase and aldosterone synthase were immunolocalised in paraffin-embedded rat adrenal and brain sections using mouse monoclonal antibodies. Negative controls utilised a mouse monoclonal antibody raised against a non-mammalian epitope. In the brain, 11beta-hydroxylase and aldosterone synthase were detected in the cerebellum, especially the Purkinje cells, as well as the hippocampus. The specificities of the 11beta-hydroxylase and aldosterone synthase antibodies were confirmed by positive immunostaining of the relevant regions of the adrenal cortex. This is the first direct evidence that steroid hydroxylases involved in the final stages of corticosteroid biosynthesis are present in specific regions of the central nervous system. PMID- 10828826 TI - Transcriptional activation of transforming growth factor alpha by estradiol: requirement for both a GC-rich site and an estrogen response element half-site. AB - 17beta-Estradiol (E2) induces transforming growth factor alpha (TGFalpha) gene expression in MCF-7 cells and previous studies have identified a 53 bp (-252 to 200) sequence containing two imperfect estrogen responsive elements (EREs) that contribute to E2 responsiveness. Deletion analysis of the TGFalpha gene promoter in this study identified a second upstream region of the promoter (-623 to -549) that is also E2 responsive. This sequence contains three GC-rich sites and an imperfect ERE half-site, and the specific cis-elements and trans-acting factors were determined by promoter analysis in transient transfection experiments, gel mobility shift assays and in vitro DNA footprinting. The results are consistent with an estrogen receptor alpha (ERalpha)/Sp1 complex interacting with an Sp1(N)(30) ERE half-site ((1/2)) motif in which both ERalpha and Sp1 bind promoter DNA. The ER/Sp1-DNA complex is formed using nuclear extracts from MCF-7 cells but not with recombinant human ERalpha or Sp1 proteins, suggesting that other nuclear factor(s) are required for complex stabilization. The E2-responsive Sp1(N)(x)ERE(1/2) motif identified in the TGFalpha gene promoter has also been characterized in the cathepsin D and heat shock protein 27 gene promoters; however, in the latter two promoters the numbers of intervening nucleotides are 23 and 10 respectively. PMID- 10828827 TI - Androgen receptor localisation and turnover in human prostate epithelium treated with the antiandrogen, casodex. AB - In vitro models of normal and malignant human prostate are currently limited to a few well established cell lines that, with a single exception (LNCaP), fail to express the androgen receptor (AR) - a common characteristic of prostatic epithelium grown in culture. To investigate the molecular mechanism of action of the non-steroidal antiandrogen Casodex (bicalutamide) against wild-type AR, we have established a transient AR expression model in non-tumorigenic prostate cells of both epithelial and mesenchymal origin. In this model, both dihydrotestosterone and Casodex can effectively transport the AR protein into the nucleus of prostate cells. Whereas the natural ligand, dihydrotestosterone, stabilises the receptor, the AR is rapidly degraded at a nuclear location when the transfected cells are treated with Casodex. In contrast, whereas the mutant AR in the LNCaP line is also degraded on Casodex treatment over the same time period, its intracellular targeting is defective. PMID- 10828828 TI - Critical role of the H6-H7 loop in the conformational adaptation of all-trans retinoic acid and synthetic retinoids within the ligand-binding site of RARalpha. AB - The pleiotropic effects of the natural and synthetic retinoids are mediated by the activation of the two subfamilies of nuclear receptors, the retinoic acid receptors (RARs) and the retinoic X receptors (RXRs). At the molecular level, these events begin with the specific ligand recognition by a nuclear receptor subtype. The adaptation of ligands to the receptor binding site leads to an optimal number of interactions for binding and selectivity which justifies elucidation of the structural requirements of the ligand binding pocket. To explore the contribution of H6-H7 loop folding in the ligand-induced conformational changes explained by the mouse-trap model, four RARalpha mutants were constructed. Ligand binding and transactivation studies revealed that three residues from the H6-H7 loop (Gly(301), Phe(302) and Gly(303)) are critical for the conformational adaptation of both synthetic agonists and antagonists. Model building and analysis of both RARalpha-ATRA and RARalpha-CD367 complexes demonstrate that accommodation of CD367 results in a less tight contact of the saturated ring of this ligand with the amino acid side chains of the receptor ligand-binding pocket compared with that of ATRA. According to the flexibility of the agonists tested (ATRA>TTNPB=Am580> CD367), we observed a decrease in binding that was dependent on ligand structure rigidity. In contrast, the binding and transactivating activities of the L266A mutant confirmed the structural constraints imposed by synthetic ligands on binding affinity for the receptor and revealed that subtle local rearrangements induced by specific conformational adaptation changes result in different binding affinities. Our results illustrate the dynamic nature of the interaction between RARalpha and its ligands and demonstrate the critical role of the H6-H7 loop in the binding of both synthetic retinoid agonists and antagonists. PMID- 10828829 TI - Multiple transcripts encoding lamprey gonadotropin-releasing hormone-I precursors. AB - The cDNA encoding lamprey prepro-gonadotropin releasing hormone-I (lamprey GnRH I) has been isolated and sequenced in an agnathan, the sea lamprey, Petromyzon marinus. The lamprey GnRH-I precursor is the first identified in an ancient lineage of vertebrates and has the same overall tripartite structure as other vertebrate GnRH precursors. The amino acid sequence of lamprey GnRH-I and the processing site (Gly-Lys-Arg) are highly conserved during 500 million years of evolution with 60-70% identity compared with those of tetrapod and teleost GnRH precursors. In contrast, the GnRH associated peptide regions are markedly divergent, with less than 20% identity compared with all identified vertebrate precursors. Unlike all other known vertebrate GnRH precursors, which typically have one and in a single case two transcripts, three distinct transcripts were isolated and sequenced in lampreys. These lamprey GnRH-I transcripts, termed GAP49, GAP50 and GAP58, differed in the length of the GAP coding sequence and were demonstrated to be the products of a single gene. Analysis of the lamprey GnRH-I gene intron-2 splice junction demonstrated that alternate splicing produces the different lamprey GnRH-I transcripts. Lamprey GnRH-I is the first GnRH gene demonstrated to utilize splice sequence variants to produce multiple transcripts, which may reflect an ancestral gene regulatory mechanism. PMID- 10828830 TI - Rapid hormonal regulation of N-acetylglucosamine transferase I. AB - Previous studies have shown that, in unstimulated mammary epithelial cells from virgin mice, prolactin receptors are retained intracellularly because of their incomplete N-glycosylation. Activation of the nitric oxide/cGMP pathway stimulates Nacetylglucosamine (NAG) transferase I activity, completion of terminal glycosylation, and redistribution of the receptors to the cell surface. In this study, it was shown that nitric oxide could stimulate the phosphorylation of NAG transferase I in intact cells and that the cGMP-dependent protein kinase (PKG) could directly phosphorylate the purified enzyme. Furthermore, this modification was associated with enhanced enzymatic activity. Conversely, this stimulation of activity was blocked in intact cells by coincubation with a PKG inhibitor and reversed in the immunoprecipitated enzyme by alkaline phosphatase treatment. Kinetic analysis revealed that this effect on enzyme activity was due to an increase in V(max) without any change in K(m). Therefore, it appears that the nitric oxide/cGMP pathway activates NAG transferase I via direct phosphorylation by PKG. PMID- 10828831 TI - TSH controls Ref-1 nuclear translocation in thyroid cells. AB - Ref-1 (called also APE) is a bifunctional protein playing a role in a large variety of cell functions. It is a major member of the DNA base excision repair system. Moreover, through reduction of cysteine residues, Ref-1 controls the activity of several transcription factors. It has been previously demonstrated that TSH up-regulates Ref-1 gene expression in thyroid cells. By using the rat FRTL-5 cell line, we demonstrate that TSH controls Ref-1 intracellular localization. Western blot experiments indicate that addition of TSH to the culture medium increases the Ref-1 cytoplasm-to-nucleus translocation. This phenomenon occurs at early times of TSH stimulation and is not dependent on protein neosynthesis. The Ref-1 cellular compartmentalization was also investigated in human thyroid tumors. A Ref-1 nuclear/cytoplasmic ratio difference between normal and cancerous thyroid tissues was observed. These results suggest that Ref-1 localization may have a critical role in the control of thyroid cell functions. PMID- 10828832 TI - Opposite regulation of brain and C-type natriuretic peptides in the streptozotocin-diabetic cardiopathy. AB - C-type natriuretic peptide (CNP), a recent addition to the family of natriuretic peptides including atrial and brain natriuretic peptide (ANP, BNP), is believed to be an endothelium-derived vasodilator and to have an antimitotic effect. ANP and BNP concentrations are increased in conditions such as congestive heart failure, but cardiac CNP concentrations have not been investigated in this connection. Diabetes mellitus also involves myocardial dysfunctions without coronary artery disease or systemic hypertension. We therefore investigated the cardiac expression of CNP mRNA compared with that of BNP mRNA in streptozotocin (STZ)-diabetic rats. STZ- diabetic male Wistar rats (n=6) were studied in comparison with controls (n=6). The animals were characterised by their mean arterial blood pressure and plasma glucose concentrations. After extraction of total cardiac RNA, a specific cDNA probe of BNP was used for northern blot analysis, whereas myocardial CNP expression was analysed by an RNase-protection assay. Twelve weeks after diabetes was induced, the rats were normotensive (96.4+/-2.0 compared with 95.1+/-1.9 mmHg) and hyperglycaemic (615+/-61 compared with 165+/-21 mg/dl; P<0.001). Left ventricular pressure was significantly impaired (76.8+/-6.4 compared with 51.2+/-3.6 mmHg). STZ-diabetic rats had a 3.2 fold increase in cardiac BNP expression compared with controls. In contrast, cardiac CNP mRNA concentrations were decreased 2.6-fold. CNP seems to be downregulated like other peptides with antimitotic and vasodilator activities (nitric oxide, prostacyclin, kinins). This may contribute to cardiac dysfunction in diabetes mellitus and suggests that stimulation of CNP expression could provide cardiac protection in such cases. PMID- 10828833 TI - Increased expression of the mRNA encoding the somatostatin receptor subtype five in human colorectal adenocarcinoma. AB - Numerous studies have suggested that the antiproliferative potency of somatostatin (SS) analogues may be an efficient tool to improve the prognosis of colorectal cancer. In order to facilitate current efforts to design potent antitumour SS analogues, we studied the distribution of human SS receptors (hsst1 5) mRNAs in a large set of tumoural and normal colonic tissues. Localisation of hsst1-5 mRNAs in normal and tumoural tissues was performed by in situ hybridisation using radioactive antisense or sense riboprobes. Semi-quantitative analysis of hsst5 mRNA was performed using a computerised image analysis system. Hsst binding sites were characterised by studying the relative potency of SS14, SS28 or SS analogues in displacing [(125)I]Tyr degrees -d-Trp(8)-SS14 bound to HT29-D4 cells. Hsst5 mRNA was by far the most expressed subtype in both normal and transformed epithelial cells as well as in the HT29-D4 cell line. An increased expression of hsst5 mRNA was found in tumours. Hsst1 mRNA was expressed preferentially as clusters in immune cells in lamina propria and in stroma close to the tumour. A low expression of hsst4, hsst3 and hsst2 was seen in normal and tumoural tissue. In HT29-D4, binding experiments with SS14 demonstrated the existence of one SS binding class (K(d)=524 nM, B(max)=1fmol/10(6 )cells). In competition binding studies, SS28 and BIM23268 (an analogue that shows preferential specificity towards hsst5) effectively inhibited binding of [(125)I]Tyr degrees -d-Trp(8)-SS14 (IC(50)=15 and 157 nM respectively), while BIM23197 (an analogue that shows preferential affinity for hsst2) was ineffective. Our results show a high expression of hsst5 mRNA in human tumoural colonic tissue, while hsst5 protein is the predominant hsst protein subtype in a tumoural colonic cell line. PMID- 10828834 TI - Cloning and regulation of the rat activin betaE subunit. AB - Using a combination of polymerase chain reaction (PCR) procedures, we have cloned and sequenced the rat activin beta(E) subunit cDNA. The putative protein corresponding to the prepro-activin beta(E) subunit was predicted to comprise 350 amino acids which, when cleaved between amino acid residues 236 and 237, would yield a mature polypeptide of approximately M(r) 12 500 with a predicted pI of 5.1. Two cDNA transcripts for activin beta(E) were identified; these differed by 738 bp in the 3'-untranslated region. Activin beta(E) mRNA transcripts were expressed only in rat liver and lung tissue as assessed by Northern blotting and PCR analysis. Relatively higher levels of both transcripts were found in the liver, whereas the lung contained lower levels that were detectable by PCR only. In situ hybridisation data showed that, within the liver, activin beta(E) mRNA was localised to hepatocytes. In vivo treatment with lipopolysaccharide as a means of activating the immune system and the hepatic acute-phase response resulted in stimulated activin beta(E) mRNA levels, compared with untreated, control rats. This increased expression was accompanied by a preferential increase in the amount of the long activin beta(E) transcript over the shorter transcript. These findings suggested that the two activin beta(E) mRNA transcripts may be products of alternative splicing events or use alternative polyadenylation sites which are differentially regulated during inflammation. These data provide evidence of a role for activin beta(E) in liver function and inflammation in the rat. PMID- 10828835 TI - The juxtamembrane but not the carboxyl-terminal domain of the insulin receptor mediates insulin's metabolic functions in primary adipocytes and cultured hepatoma cells. AB - Insulin-stimulated signaling pathways are activated upon interactions between the intracellular domains of the receptor and its downstream effectors. Insulin receptor substrate proteins (IRS-1, -2, -3 and -4) are the best-studied substrates for the insulin receptor kinase (IRK). We have previously shown that IRS-1 and IRS-2 interact with the juxtamembrane (JM) but not with the carboxyl terminal (CT) region of the insulin receptor (IR) in vitro. However, the precise role of these IR regions in mediating insulin's bioeffects is still unresolved. In the present work we made use of vaccinia virus as a vector for quantitative expression of the JM and CT domains within the cytoplasm of physiologically insulin-responsive primary rat adipocytes and rat hepatoma Fao cells. We could demonstrate that overexpression of either the JM or the CT domains did not inhibit either insulin binding or insulin-stimulated receptor autophosphorylation. In contrast, metabolic effects such as insulin-induced glucose utilization in adipocytes, and insulin-induced amino acid utilization in Fao hepatoma cells were inhibited (70-80%) in cells overexpressing the JM but not the CT domains of IR. The inhibitory effects of the overexpressed JM domain were accompanied by inhibition of insulin-stimulated IRS-1 phosphorylation, decreased IRS-1-associated PI3K activity, and decreased phosphorylation of the downstream effectors of PI3K, PKB and p70 S6K. Insulin-stimulated thymidine incorporation in Fao cells was also inhibited (40%) upon overexpression of the JM but not the CT region of IR. Our findings suggest that interactions between the JM region of IR and its downstream effectors are obligatory for insulin-stimulated metabolic functions in physiologically relevant insulin responsive cells. They also rule out the possibility that interaction of proteins, including PI3K, with the CT domain can provide an alternative pathway. PMID- 10828836 TI - Semiquantitative reverse transcription-polymerase chain reaction to evaluate the expression patterns of genes involved in the oestrogen pathway. AB - The increasing number of factors to be taken into account in the oestrogen transcriptional process has created a need to develop a rapid screening method to evaluate their role in physiology and pathology. Molecular biology techniques enable gene expression studies at the mRNA level with small amounts of tissues. We therefore developed a semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) technique using fluorescent oligonucleotides to analyse simultaneously a large panel of interrelated genes involved in the oestrogen transcriptional pathway using a moderately expressed housekeeping gene, the hypoxanthine phosphoribosyltransferase gene (HPRT), as the reference gene. Expression levels of oestrogen receptors (ERalpha, ERbeta), cofactors AIB1, RIP140, SMRT and the Fas-associated protein-tyrosine phophatase-1 (FAP-1) genes were evaluated in breast, endometrial and ovarian cancer cell lines and in three ERalpha-positive and three ERalpha-negative breast cancer tumours. This technique provides a rapid and reliable way to quantitate simultaneously numerous mRNAs of genes involved in the oestrogen pathway from small amounts of tissues. PMID- 10828837 TI - IGF-I and IGF-I-binding proteins in rats with adjuvant-induced arthritis given recombinant human growth hormone. AB - Adjuvant-induced arthritis in rats is associated with growth failure, hypermetabolism and accelerated protein breakdown. We have previously reported that adjuvant-induced arthritis in rats results in a decrease in body weight gain, pituitary GH mRNA, circulating GH and IGF-I together with an increase in serum IGF-binding proteins (IGFBPs). The aim of this study was to analyze the role of GH in the decrease in body weight and in the alterations in the IGF-I system observed in chronic inflammation. Male Wistar rats were injected with complete Freund's adjuvant and 16 days later arthritic rats were injected daily with recombinant human GH (rhGH) (3 IU/kg s.c.) for 8 days; control rats received 250 microl saline. Arthritis significantly decreased body weight gain and serum IGF-I. These decreases were not due to the reduced food intake, since in pair-fed rats they were not observed. Furthermore, administration of rhGH to arthritic rats increased body weight gain without modifying food intake. To further investigate the effect of GH administration, 14 days after adjuvant injection both control and arthritic rats were treated with 0, 1.5, 3 or 6 IU/kg of rhGH. GH treatment at the dose of 3 and 6 IU/kg significantly increased body weight gain in arthritic rats. GH administration, at the higher dose of 6 IU/kg, increased hepatic and serum concentrations of IGF-I in both control and arthritic rats. In control rats, rhGH at the three doses assayed increased circulating IGFBP-3. GH treatment in arthritic rats decreased IGFBP-1 and -2, and did not modify IGFBP-4. GH treatment at the dose of 3 IU/kg also decreased circulating IGFBP-3 in arthritic rats. These data suggest that GH treatment can ameliorate the catabolism observed in adjuvant-induced arthritis, an effect mediated, at least in part, by modifications in the circulating IGFBPs. PMID- 10828838 TI - Alterations in insulin-like growth factor binding protein-3 proteolysis and complex formation in the arthritic joint. AB - Increased concentrations of insulin-like growth factor (IGF) system components have previously been observed in rheumatoid arthritis (RA) and osteoarthritis (OA); however, disruption of the IGF axis and the implications for the disease process remain largely unaddressed. This study was undertaken to characterise the IGF binding protein (IGFBP)-3 proteolysis and complex formation systems in synovial fluid and to investigate changes in these systems in arthritic disease, and their impact on the availability of IGF. Western blotting or autoradiography of SDS gels was used to visualise IGFBP-3 or its proteolysis. IGF-I and IGFBP-3 concentrations were determined by radioimmunoassays and acid-labile subunit (ALS) was measured by ELISA. A shift in distribution of IGFBP-3 and IGF-I in RA and OA synovial fluids (RASynF, OASynF) and an associated increase in ALS suggested the presence of 150 kDa ternary complexes. IGFBP-3 proteolysis was decreased in RASynF and OASynF, but was apparent in size-fractionated fluid and resembled serum activity. The presence of serum-like inhibitors of IGFBP-3 proteolysis in RASynF was also demonstrated by the ability of this fluid, and 150 kDa fractions from its size fractionation, to inhibit IGFBP-3 proteolysis in the other synovial fluid. A marked disruption in the IGF system was observed, as considerably more IGF-I was retained in ternary complexes. We also classified the IGFBP-3 proteolysis system in synovial fluid and found it to be disturbed in RASynF and OASynF. These changes may be caused by an increased flux of circulatory proteins into synovial fluid, resulting from an inflammation-induced increase in vascular permeability. The net result in RA and OA would be a decrease in IGF availability in arthritic joints, and therefore loss of a potential anabolic stimulus. This disruption to the IGF axis would influence disease progression in RA and OA. PMID- 10828839 TI - Differential regulation of IGF-binding proteins in rabbit costal chondrocytes by IGF-I and dexamethasone. AB - Cartilage is a primary target tissue for the IGFs. The mitogenic activity of these peptides is regulated by a family of high-affinity IGF-binding proteins (IGFBP-1 to -6). We characterized the IGFBPs produced by cultured chondrocytes derived from rib cartilage of prepubertal rabbits. Culture medium, which had been conditioned by these cells for 48 h showed bands of 22 kDa, 24 kDa and a 31/32 kDa doublet by Western ligand blotting with [(125)I]IGF-II. When the cells were grown in the presence of increasing amounts of IGF-I or IGF-II, the 31/32 kDa doublet increased in intensity (reaching a plateau of about 11-fold stimulation between 2 and 10 nM IGF-I). The 22 kDa and 24 kDa bands increased only slightly while a 26 kDa band became faintly visible. By Western immunoblotting the 31/32 kDa doublet was identified as IGFBP-5. An IGF-I analog with reduced affinity for IGFBPs, Long-R3 IGF-I, also induced IGFBP-5, while insulin was less effective (2.2-fold stimulation at 10 nM). IGF-I protected IGFBP-5 against proteolytic degradation by conditioned medium. IGF-I also enhanced the level of IGFBP-5 mRNA. LY294002, a specific inhibitor of the intracellular signaling molecule phosphatidylinositol 3-kinase, inhibited stimulation of IGFBP-5 by IGF-I. Dexamethasone suppressed IGFBP-5 (by 70% at 20 nM) but, at the same time, a 39/41 kDa doublet (presumably IGFBP-3) was induced. IGFBP-5 has been shown in several cell types to enhance the mitogenic activity of IGF-I. IGFBP-3 generally acts as a growth inhibitor. Therefore, the differential effects of dexamethasone on these regulatory proteins could account, at least in part, for the growth-arresting effect of this glucocorticoid. PMID- 10828840 TI - The GH secretagogues ipamorelin and GH-releasing peptide-6 increase bone mineral content in adult female rats. AB - Growth hormone (GH) is of importance for normal bone remodelling. A recent clinical study demonstrated that MK-677, a member of a class of GH secretagogues (GHSs), increases serum concentrations of biochemical markers of bone formation and bone resorption. The aim of the present study was to investigate whether the GHSs, ipamorelin (IPA) and GH-releasing peptide-6 (GHRP-6), increase bone mineral content (BMC) in young adult female rats. Thirteen-week-old female Sprague-Dawley rats were given IPA (0.5 mg/kg per day; n=7), GHRP-6 (0.5 mg/kg per day; n=8), GH (3.5 mg/kg per day; n=7), or vehicle administered continuously s.c. via osmotic minipumps for 12 weeks. The animals were followed in vivo by dual X-ray absorptiometry (DXA) measurements every 4th week. After the animals were killed, femurs were analysed in vitro by mid-diaphyseal peripheral quantitative computed tomography (pQCT) scans. After this, excised femurs and vertebrae L6 were analysed by the use of Archimedes' principle and by determinations of ash weights. All treatments increased body weight and total tibial and vertebral BMC measured by DXA in vivo compared with vehicle-treated controls. However, total BMC corrected for the increase in body weight (total BMC:body weight ratio) was unaffected. Tibial area bone mineral density (BMD, BMC/area) was increased, but total and vertebral area BMDs were unchanged. The pQCT measurements in vitro revealed that the increase in the cortical BMC was due to an increased cross sectional bone area, whereas the cortical volumetric BMD was unchanged. Femur and vertebra L6 volumes were increased but no effect was seen on the volumetric BMDs as measured by Archimedes' principle. Ash weight was increased by all treatments, but the mineral concentration was unchanged. We conclude that treatment of adult female rats with the GHSs ipamorelin and GHRP-6 increases BMC as measured by DXA in vivo. The results of in vitro measurements using pQCT and Archimedes' principle, in addition to ash weight determinations, show that the increases in cortical and total BMC were due to an increased growth of the bones with increased bone dimensions, whereas the volumetric BMD was unchanged. PMID- 10828841 TI - An osteogenesis-related transcription factor, core-binding factor A1, is constitutively expressed in the chondrocytic cell line TC6, and its expression is upregulated by bone morphogenetic protein-2. AB - Core-binding factor A1 (Cbfa1), also called Pebp2 alpha A/AML3, is a transcription factor that belongs to the runt-domain gene family. Cbfa1-deficient mice are completely incapable of both endochondral and intramembranous bone formation, indicating that Cbfa1 is indispensable for osteogenesis. Maturation of chondrocytes in these mice is also disorganized, suggesting that Cbfa1 may also play a role in chondrogenesis. The aim of this study was to examine the expression and regulation of Pebp2 alpha A/AML3/Cbfa1 expression in the chondrocyte-like cell line, TC6. Northern blot analysis indicated that Cbfa1 mRNA was constitutively expressed as a 6.3 kb message in TC6 cells and the level of Cbfa1 expression was enhanced by treatment with bone morphogenetic protein-2 (BMP2) in a time- and dose-dependent manner. This effect was blocked by an RNA polymerase inhibitor, 5,6-dichloro-1-beta-d-ribofuranosylbenzimidazole, but not by a protein synthesis inhibitor, cycloheximide. Western blot analysis of the cell lysates using polyclonal antibody raised against Cbfa1 indicated that BMP2 treatment increased the Cbfa1 protein level in TC6 cells. In TC6 cells, BMP2 treatment enhanced expression of alkaline phosphatase and type I collagen mRNAs but suppressed that of type II collagen mRNA. In addition to TC6 cells, Cbfa1 mRNA was also expressed in primary cultures of chondrocytes and BMP2 treatment enhanced Cbfa1 mRNA expression in these cells similarly to its effect on TC6 cells. These data indicate that the Pebp2 alpha A/AML3/Cbfa1 gene is expressed in a chondrocyte-like cell line, TC6, and its expression is enhanced by treatment with BMP. PMID- 10828842 TI - Peptidomimetic antagonists of alphavbeta3 inhibit bone resorption by inhibiting osteoclast bone resorptive activity, not osteoclast adhesion to bone. AB - Osteoclasts are actively motile on bone surfaces and undergo alternating cycles of migration and resorption. Osteoclast interaction with the extracellular matrix plays a key role in the osteoclast resorptive process and a substantial body of evidence suggests that integrin receptors are important in osteoclast function. These integrin receptors bind to the Arg-Gly-Asp (RGD) sequence found in a variety of extracellular matrix proteins and it is well established that the interaction of osteoclast alpha v beta 3 integrin with the RGD motif within bone matrix proteins is important in osteoclast-mediated bone resorption. In this study, we characterized the effects of two synthetic peptidomimetic antagonists of alpha v beta 3, SC-56631 and SC-65811, on rabbit osteoclast adhesion to purified matrix proteins and bone, and on bone resorption in vitro. SC-56631 and SC-65811 are potent inhibitors of vitronectin binding to purified alpha v beta 3. Both SC-56631 and SC-65811 inhibited osteoclast adhesion to osteopontin- and vitronectin-coated surfaces and time-lapse video microscopy showed that osteoclasts rapidly retract from osteopontin-coated surfaces when exposed to SC 56631 and SC-65811. SC-56631 and SC-65811 blocked osteoclast-mediated bone resorption in a dose-responsive manner. Further analysis showed that SC-65811 and SC-56631 reduced the number of resorption pits produced per osteoclast and the average pit size. SC-65811 was a more potent inhibitor of bone resorption and the combination of reduced pit number and size led to a 90% inhibition of bone resorption. Surprisingly, however, osteoclasts treated with SC-65811, SC-56631 or the disintegrin echistatin, at concentrations that inhibit bone resorption did not inhibit osteoclast adhesion to bone. These results suggest that alphavbeta3 antagonists inhibited bone resorption by decreasing osteoclast bone resorptive activity or efficiency but not by inhibiting osteoclast adhesion to bone per se. PMID- 10828843 TI - Thyroid hormone uptake in cultured rat anterior pituitary cells: effects of energy status and bilirubin. AB - Transport of thyroxine (T(4)) into the liver is inhibited in fasting and by bilirubin, a compound often accumulating in the serum of critically ill patients. We tested the effects of chronic and acute energy deprivation, bilirubin and its precursor biliverdin on the 15-min uptake of [(125)I]tri-iodothyronine ([(125)I]T(3)) and [(125)I]T(4) and on TSH release in rat anterior pituitary cells maintained in primary culture for 3 days. When cells were cultured and incubated in medium without glucose and glutamine to induce chronic energy deprivation, the ATP content was reduced by 45% (P<0. 05) and [(125)I]T(3) uptake by 13% (NS), but TSH release was unaltered. Preincubation (30 min) and incubation (15 min) with 10 microM oligomycin reduced ATP content by 51% (P<0.05) and 53% (P<0. 05) under energy-rich and energy-poor culture conditions respectively; [(125)I]T(3) uptake was reduced by 66% (P<0.05) and 64% (P<0.05). Neither bilirubin nor biliverdin (both 1-200 microM) affected uptake of [(125)I]T(3) or [(125)I]T(4). Bilirubin (1-50 microM) did not alter basal or TRH-induced TSH release. In conclusion, the absence of inhibitory effects of chronic energy deprivation and bilirubin on thyroid hormone uptake by pituitary cells supports the view that the transport is regulated differently than that in the liver. PMID- 10828844 TI - Tissue-specific regulation of thyroid hormone receptor mRNA isoforms and target gene proteins in domestic ducks. AB - Skeletal muscles are important target tissues for thyroid hormone action. The present study examines the influence of thyroid status on muscle growth and tissue-specific expression of thyroid receptor (TR) mRNA isoforms in a commercial strain of the domestic duck (Anas platyrhynchos). Four groups (n=5) of 1-week-old ducklings were rendered either hypothyroid by treatment with methimazole (6 mg 100 g(-1) body mass or 12 mg 100 g(-1) body mass), or hyperthyroid by treatment with methimazole (6 mg 100 g(-1) body mass) in combination with thyroid hormones (5 microg thyroxine (T(4)) and tri-iodothyronine (T(3)) 100 g(-1) body mass or 10 microg T(4) and T(3) 100 g(-1) body mass). Serum and tissue samples (cardiac, pectoralis and semimembranosus leg muscle, liver, pituitary and cerebral cortex) were collected from these four groups, and from a group of untreated controls, at 8 weeks of age. Development of duckling morphology was retarded in methimazole treated birds compared with that in euthyroid controls, as evidenced by differences in skeletal dimensions, primary feather length, and body and muscle masses. Body mass was lower by 18%, and relative masses of cardiac and pectoralis muscles were lower by 28% and 32% respectively. Heterologous oligonucleotides for TR alpha, TR beta 0, TR beta2 and the housekeeping gene beta-actin were derived from chicken sequences. RT-PCR showed that TR alpha mRNA was expressed in all tissues but was not significantly affected by any of the experimental treatments. TR beta 0 mRNA expression was significantly lower in the leg muscles of ducklings treated with 12 mg methimazole 100 g(-1) body mass (0.109+/-0.047 TR:beta-actin ratio, P<0.05) compared with that in euthyroid controls (0.380+/-0.202), but was unaltered in the pectoralis and cardiac muscles. Expression of TR beta 0 mRNA was significantly higher in pectoralis (by 3.5-fold, P<0. 05), cardiac (by 4.2-fold, P=0.003) and leg (by 4.0-fold, P<0.001) muscles of ducklings treated with thyroid hormones compared with those in euthyroid controls (0.098+/-0.019, 0.822+/-0.297 and 0. 38+/-0.202 TR:beta-actin respectively). Only the pituitary gland expressed significant levels of TR beta 2 mRNA. PMID- 10828845 TI - Expression of vascular endothelial growth factor in response to high glucose in rat mesangial cells. AB - Diabetic nephropathy associated with hyperglycemia is characterized by glomerular hyperfiltration and endothelial dysfunction. Vascular endothelial growth factor (VEGF) is known to be primarily involved in neoangiogenesis and increased endothelial permeability. The purpose of this study was to investigate VEGF expression in response to high glucose in rat cultured mesangial cells and to identify its signal pathway via protein kinase C (PKC). Rat mesangial cells were cultured with different concentrations of glucose: normal (5 mM d-glucose), medium (15 mM d-glucose) and high (30 mm d-glucose). Calphostin-C as a PKC inhibitor and phorbol myristate acetate (PMA) as a PKC downregulator were instillated into culture media to evaluate the role of PKC in mediating the glucose-induced increase in VEGF expression. High glucose increased expression of VEGF at the mRNA and protein levels, identified by semi-quantitative RT-PCR and western blotting, within 3 h and in a time- and glucose concentration-dependent manner. Calphostin-C and PMA inhibited glucose-induced increases in VEGF expression at the mRNA and protein levels. In conclusion, high glucose can directly increase VEGF expression in rat mesangial cells via a PKC-dependent mechanism. These results suggest that VEGF could be a potential mediator of glomerular hyperfiltration and proteinuria in diabetic nephropathy. PMID- 10828846 TI - Level of nutrition affects leptin concentrations in plasma and cerebrospinal fluid in sheep. AB - In mature male sheep, the level of nutrition acutely influences the secretion of reproductive hormones. The mechanism involved is not fully understood but findings in humans and laboratory rodents would suggest a major role for leptin that is secreted from adipose tissue and then travels via the circulation to the central nervous system. Before we can begin to test this hypothesis, we need to be able to measure leptin concentrations in blood plasma and cerebrospinal fluid. We have therefore developed a radioimmunoassay using antibodies raised against biologically active recombinant bovine-ovine leptin. Using this assay, we found that plasma concentrations of leptin were highly correlated to back-fat thickness and to the ratio of back-fat thickness to liveweight, in female and castrated male sheep. Plasma concentrations of leptin were higher in female sheep than in castrated or intact male sheep. Serial samples (every 5 min) suggested that the secretion of leptin in male sheep is episodic but it does not appear to show clear pulsatility, increases post-prandially, or a diurnal rhythm. Leptin concentrations in both plasma and cerebrospinal fluid increased within 5 days in male sheep fed a diet with a high content of energy and protein that also stimulates the secretion of LH pulses. These data suggest that in sheep, as in other species, leptin production is correlated with the mass of adipose tissue and that the hormone passes from the circulation to the cerebrospinal fluid and then to hypothalamic sites. There, it may affect appetite and perhaps GnRH secretion. The role of leptin in the link between nutrition and reproduction needs further investigation. PMID- 10828847 TI - Improved glycaemic control in obese diabetic ob/ob mice using N-terminally modified gastric inhibitory polypeptide. AB - Gastric inhibitory polypeptide (GIP) is an important insulin-releasing hormone of the enteroinsular axis which is rapidly inactivated by the exopeptidase dipeptidyl peptidase (DPP) IV. The present study has examined the ability of Tyr(1)-glucitol GIP to be protected from plasma degradation and to enhance insulin-releasing and antihyperglycaemic activity in 20- to 25-week-old obese diabetic ob/ob mice. Degradation of GIP by incubation at 37 degrees C with obese mouse plasma was clearly evident after 3 h (35% degraded). After 6 h, more than 61% of GIP was converted to GIP(3-42) whereas N-terminally modified Tyr(1) glucitol GIP was resistant to degradation in plasma (>99% intact after 6 h). The formation of GIP(3-42) was almost completely abolished by inhibition of plasma DPP IV with diprotin A. Effects of GIP and Tyr(1)-glucitol GIP were examined in overnight-fasted obese mice following i.p. injection of either peptide (20 nmol/kg) together with glucose (18 mmol/kg) or in association with feeding. Most prominent effects were observed in the former group where plasma glucose values at 60 min together with the area under the curve (AUC) for glucose were significantly lower following GIP (AUC, 874+/-72 mmol/l.min; P<0.01) or Tyr(1) glucitol GIP (770+/-134 mmol/l.min; P<0.001) as compared with administration of glucose alone (1344+/-136 mmol/l.min). This was associated with a significantly greater and more protracted insulin response following Tyr(1)-glucitol GIP than GIP (AUC, 491+/-118 vs 180+/-33 ng/ml.min; P<0.05). Administration of Tyr(1) glucitol GIP also enhanced the glucose-lowering ability of 50 units/kg insulin (218.4+/-30.2 vs insulin alone 133.9+/-16.2 mmol/l.min; P<0.05). These data demonstrate that Tyr(1)-glucitol GIP displays resistance to plasma DPP IV degradation in a commonly used animal model of type 2 diabetes, resulting in enhanced antihyperglycaemic activity and insulin-releasing action in vivo. PMID- 10828848 TI - Plasma cholesteryl ester fatty acid composition, insulin sensitivity, the menopause and hormone replacement therapy. AB - This study was designed to determine the effect of menopause and hormone replacement therapy (HRT) on plasma cholesteryl ester fatty acid (CEFA) composition and insulin sensitivity and the relationships between these variables in perimenopausal women (aged 40-55 years) including 49 who were premenopausal and 32 who were postmenopausal. Plasma cholesteryl ester proportions of dihomo gamma-linolenic acid (20:3 n-6) were correlated significantly with insulin sensitivity index (r=-0.319, P=0.005), fasting serum insulin levels (r=0.230, P=0.038), body mass index (r=0.242, P=0.03) and per cent body fat (r=0.329, P=0.003) in perimenopausal women (n=81). Similar associations were observed in premenopausal women. Regression analysis suggested the relationships between 20:3 n-6 proportions and indices of insulin action may be partly mediated by levels of adiposity. In postmenopausal women, 6 months of HRT significantly (P=0.008) increased the ratio of arachidonic acid (20:4 n-6) to linoleic acid (18:2 n-6), which is an indicator of activity in the pathway of 20:4 n-6 synthesis, compared with placebo. These findings suggest that the type of fat in the diet indicated by plasma CEFA composition is linked to adiposity and insulin action. They also suggest that in postmenopausal women, HRT may increase the synthesis of 20:4 n-6, which is the precursor for eicosanoids with important cardiovascular functions. PMID- 10828849 TI - Parathyroid hormone-related protein(1-34) in gestational fluids and release from human gestational tissues. AB - Parathyroid hormone-related protein (PTHrP) is present in fetal and gestational tissues, in which its proposed roles include stimulation of epithelial growth and differentiation, vasodilatation of the uteroplacental vasculature, relaxation of uterine muscle and stimulation of placental calcium transport. The aim of this study was to determine whether the release of PTHrP from gestational tissue explants was tissue specific. In addition, PTHrP concentrations were measured in maternal plasma, umbilical artery and vein plasma, and amniotic fluid from term, uncomplicated pregnancies before the onset of labour. PTHrP was detected in low concentrations in the mother, fetus and placental tissue. Amniotic fluid had ten times the PTHrP concentration compared with that in the maternal or fetal circulations. Using late pregnant human gestational tissues in an in vitro explant system, we found that amnion over placenta, choriodecidua, reflected amnion, and placenta released PTHrP into culture medium in progressively greater amounts over 24 h (P<0.05). This release was not associated with a loss of cell membrane integrity, as indicated by measurement of the intracellular enzyme, lactate dehydrogenase, in the incubation media. After 24 h incubation, the fetal membranes released significantly (P<0.05) greater amounts of PTHrP than did the placenta (placenta 3. 7+/-0.5 pmol PTHrP/g protein). Amnion over placenta released significantly more PTHrP (139.3+/- 43.1 pmol PTHrP/g protein) than did reflected amnion (29.0+/-8.3 pmol PTHrP/g protein) (P<0.05). This study unequivocally demonstrated that human gestational tissues release PTHrP and it was concluded that the main contributors to PTHrP in amniotic fluid were the human fetal membranes, particularly amnion over placenta. Fetal membrane-derived and amniotic fluid PTHrP are proposed to have stimulatory effects on epithelial growth and differentiation in fetal lung, gut, skin and hair follicles and paracrine effects on placental vascular tone and calcium transport. PMID- 10828850 TI - Amylin inhibits ovariectomy-induced bone loss in rats. AB - Amylin (AMY), a peptide co-secreted with insulin by pancreatic beta-cells, inhibits bone resorption and stimulates osteoblastic activity. The ovariectomized (OVX) rat is an established animal model for human osteoporosis. Thus, the present experiment was performed to study the effects of AMY on estrogen deficiency-induced bone loss in rats. Thirty-one 6-month-old Wistar rats were randomized by body weight (BW) into two groups. The first underwent surgical OVX (n=21). The second was sham-operated (SH; n=10). Sixty days after surgery, 11 OVX rats were s.c. injected with rat AMY (3 microg/100 g BW/day, for 30 days; OVX+AMY), and 10 with solvent alone in the same way (0.15 ml/100 g BW; OVX). Each rat, housed in an individual cage, was fed daily the mean quantity of diet consumed the day before by SH rats. This diet contained 0.24% calcium and 0. 16% phosphorus. The 31 animals were killed on day 90. No difference in daily weight gain and BW was observed between groups. Neither AMY treatment nor OVX had any significant effect upon femoral morphology, femoral failure load, diaphyseal femoral density (representative of cortical bone) and total femoral calcium content. Nevertheless, both distal metaphyseal (representative of cancellous bone) and total femoral bone densities were higher in SH and OVX+AMY than in OVX rats. The highest plasma osteocalcin concentration was measured in OVX+AMY rats. Simultaneously, urinary deoxypyridinoline excretion was lower in OVX+AMY than in OVX rats. These results indicate that in OVX rats, AMY treatment inhibited trabecular bone loss both by inhibiting resorption and by stimulating osteoblastic activity. PMID- 10828851 TI - Radioimmunoassay of plasma ouabain in healthy and pregnant individuals. AB - Ouabain was recently isolated from human plasma, bovine hypothalamus and bovine adrenal in attempts to identify endogenous substances inhibiting the cell membrane sodium pump. A number of radioimmunoassays have been developed in order to study the clinical significance of ouabain. The results have been controversial with regard to the presence and chemical nature of plasma ouabain like immunoreactivity. We have now measured ouabain in healthy and pregnant individuals using solid-phase extraction of plasma samples followed by a new radioimmunoassay with the extraordinary sensitivity of at least 2 fmol/tube (5 pmol/l). Plasma extracts, a previously isolated human plasma ouabain-like compound and bovine hypothalamic inhibitory factor displaced the tracer in parallel and eluted identically with ouabain in high-performance liquid chromatography. Plasma ouabain immunoreactivity was found to be much lower than reported previously: 12.6+/-1.3 pmol/l in healthy men (mean+/-s.e., n=20) and 9.4+/-0.7 pmol/l in women (n=14). In pregnant women (n=28) plasma ouabain concentration was 16.3+/-4.0 pmol/l during the first trimester, 18.8+/-4.3 pmol/l during the second trimester and 24.3+/-4.0 pmol/l during the third trimester (all P<0.01 compared with non-pregnant women). Plasma ouabain 3-5 days after the delivery was 13.6+/-1.1 pmol/l (n=10, P<0.05-0.01 compared with second and third trimesters). The pregnancy-related changes in the plasma concentrations of ouabain resembled those of cortisol. Therefore cortisol was measured from the same plasma samples and a significant positive correlation was found (r=0.512, P=0.006). The similar profiles of plasma ouabain and cortisol during pregnancy and their rapid decreases postpartum are consistent with the adrenal cortical origin of ouabain and also show that the secretions of these hormones are possibly under the control of same factors. PMID- 10828852 TI - Hormonal diagnosis of 21-hydroxylase deficiency in plasma and urine of neonates using benchtop gas chromatography-mass spectrometry. AB - We aimed at measuring the first plasma concentrations of 17-hydroxyprogesterone (17OH-P) determined by benchtop isotope dilution/gas chromatography-mass spectrometry (ID/GC-MS) in term neonates with or without 21-hydroxylase deficiency. Plasma samples from normal cord blood specimens (n=30), unaffected neonates (n=38) and neonatal patients with classical 21-hydroxylase deficiency (eight salt-wasters, three simple virilizers) were analyzed. Steroid profiling of random urinary specimens by GC-MS served as a confirmatory test for 21 hydroxylase deficiency. 17OH-P (nmol/l) in cord blood plasma lay between 11.66 and 75.92 (median 24.74). It declined shortly after birth. In the first 8 days of life, the time that screening for 21-hydroxylase deficiency is performed, 17OH-P ranged between undetected levels and an upper limit of 22.87 (median 4.11). Thereafter (days 9-28) its concentrations lay between 2.18 and 20.30 (median 6.22). Except one simple virilizer, all other patients with 21-hydroxylase deficiency had clearly elevated plasma 17OH-P at the time that screening for 21 hydroxylase deficiency would be performed. We suggest ID/GC-MS, which provides the highest specificity in steroid analysis, for checking suspicious concentrations of 17OH-P in neonates and underscore the potential of urinary steroid profiling by GC-MS as a rapid, non-invasive and non-selective confirmatory test for congenital adrenal hyperplasia. PMID- 10828853 TI - 11 beta-hydroxysteroid dehydrogenase type 1 is a predominant 11 beta-reductase in the intact perfused rat liver. AB - 11 beta-Hydroxysteroid dehydrogenase type 1 (11 beta-HSD-1), a regulator of intrahepatocellular glucocorticoid activity, is bidirectional in homogenates but catalyses 11 beta-reduction (regenerating glucocorticoid) in intact primary hepatocytes in culture. To examine this discrepancy at the whole-organ level, we examined 11 beta-HSD-1 activity in the intact bivascularly perfused rat liver. On a single pass through male rat liver, 44+/-5% of 11-dehydrocorticosterone (11 DHC) recovered was 11 beta-reduced to corticosterone, whereas 10+/-1% of corticosterone was 11 beta-dehydrogenated to 11-DHC. 11 beta-Reduction was less in female liver (21+/-2%, P<0.01) and was significantly greater with perfusion of all substrate via the portal vein (50+/-3%) than via the hepatic artery (30+/-2%, P<0.05). 11 beta-Reductase activity was not saturated by 11-DHC (10(-)(9)-10( )(6) M). Perfusion with carbenoxolone (CBX, 10(-)(6)-10(-)(3 )M) did not alter 11 beta-reduction of 11-DHC. In contrast, pretreatment with CBX in vivo (10 mg/day) for 7 days inhibited 11 beta-reductase (19+/-4% conversion, P<0.01). Concentrations of 11-DHC in male rat plasma were 44+/-6 nM. Thus 11 beta-HSD-1 is predominantly an 11 beta-reductase in the intact rat liver and is only inhibited by chronic administration of CBX. The substantial concentrations of plasma 11-DHC as substrate suggest that 11 beta-HSD-1 activity and its potential selective inhibition could modify glucocorticoid action in vivo. PMID- 10828854 TI - The oestrogenic effects of gestodene, a potent contraceptive progestin, are mediated by its A-ring reduced metabolites. AB - Gestodene (17 alpha-ethynyl-13 beta-ethyl-17 beta-hydroxy-4, 15-gonadien-3-one) is the most potent synthetic progestin currently available and it is widely used as a fertility regulating agent in a number of contraceptive formulations because of its high effectiveness, safety and acceptability. The observation that contraceptive synthetic progestins exert hormone-like effects other than their progestational activities, prompted us to investigate whether gestodene (GSD) administration may induce oestrogenic effects, even though the GSD molecule does not interact with intracellular oestrogen receptors (ER). To assess whether GSD may exert oestrogenic effects through some of its neutral metabolites, a series of experimental studies were undertaken using GSD and three of its A-ring reduced metabolites. Receptor binding studies by displacement analysis confirmed that indeed GSD does not bind to the ER, whereas its 3 beta,5 alpha-tetrahydro reduced derivative (3 beta GSD) interacts with a relative high affinity with the ER. The 3 alpha,5 alpha GSD isomer (3 alpha GSD) also binds to the ER, though to a lesser extent. The ability of the A-ring reduced GSD derivatives to induce oestrogenic actions was evaluated by the use of two different molecular bioassays: (a) transactivation of a yeast system co-transfected with the human ER alpha (hER alpha) gene and oestrogen responsive elements fused to the beta-galactosidase reporter vector and (b) transactivation of the hER alpha-mediated transcription of the chloramphenicol acetyl transferase (CAT) reporter gene in a HeLa cells expression system. The oestrogenic potency of 3 beta GSD was also assessed by its capability to induce oestrogen-dependent progestin receptors (PR) in the anterior pituitary of castrated female rats. The results demonstrated that 3 beta GSD and 3 alpha GSD were able to activate, in a dose-dependent manner, the hER alpha mediated transcription of both the beta-galactosidase and the CAT reporter genes in the yeast and HeLa cells expression systems respectively. In both assays the 3 beta derivative of GSD exhibited a significantly greater oestrogenic effect than its 3 alpha isomer, while unchanged GSD and 5 alpha GSD were completely ineffective. Neither 3 beta GSD nor 3 alpha GSD exhibited oestrogen synergistic actions. Interestingly, the pure steroidal anti-oestrogen ICI-182,780 diminished the transactivation induced by 3 beta GSD and 3 alpha GSD in the yeast expression system. Furthermore, administration of 3 beta GSD resulted in a significant increase of oestrogen-dependent PR in the anterior pituitaries of castrated rats in comparison with vehicle-treated animals. The characteristics of the 3 beta GSD induced PR were identical to those induced by oestradio benzoate. The overall results demonstrate that 3 beta GSD and its 3 alpha isomeric alcohol specifically bind to the ER and possess a weak intrinsic oestrogenic activity, whereas unmodified GSD does not. The data contribute to a better understanding of the GSD mechanism of action and allow the hypothesis to be advanced that the slight oestrogenlike effects attributable to GSD are mediated by its non-phenolic, tetrahydro reduced metabolites. PMID- 10828855 TI - Characterisation of N-terminal chromogranin A and chromogranin B in mammals by region-specific radioimmunoassays and chromatographic separation methods. AB - Chromogranin A (CgA) and chromogranin B (CgB) are acidic proteins stored in and released from hormone granules in endocrine and neuroendocrine tissue. The chromogranins are postulated to serve as pro-hormones to generate biologically active peptides, which may influence hormonal release and vascular functions or have antibacterial functions. Although N-terminal and C-terminal regions show some species amino acid homology, the chromogranins as a whole display considerable interspecies differences, which prevents their use in comparative studies of biological functions. We present four new radioimmunoassays for the measurement of defined N-terminal regions of CgA and CgB. A new radioimmunoassay for measurement of intact bovine CgA has also been developed. With these assays and two previously published ones, we have compared the cross-reactivity of chromogranins from man, cattle, sheep, goat, pig and horse and compared adrenomedullar content and serum levels of CgA from these species. We have also studied the influence of peptide concentrations and the ionic strength of the mobile phase on molecular weight estimations. Assays with antibodies directed against the N-terminal parts of CgA and CgB showed sufficient interspecies cross reactivity to allow comparative quantification of the circulating levels in man, cattle, sheep, goat, pig and horse. Assays measuring the intact human or bovine CgA were not suitable for comparative purposes in samples from sheep, goat, pig and horse. Molecular interactions between vasostatin immunoreactive material and intact bovine CgA were demonstrated in gel permeation studies, suggesting that conclusions about the degree of N-terminal processing from elution profiles should be made with caution. Reliable interspecies comparison of chromogranins is difficult, but measurements with region-specific assays may be helpful to study concentrations of chromogranins and chromogranin-related peptides. PMID- 10828856 TI - Progesterone on an oestrogen background enhances prolactin-induced apoptosis in regressing corpora lutea in the cyclic rat: possible involvement of luteal endothelial cell progesterone receptors. AB - Preovulatory surges of both prolactin (PRL) and progesterone have been suggested to be necessary for the induction of apoptosis in the regressing corpus luteum of the cyclic rat. The aim of these experiments was to study whether the administration of PRL and/or progesterone on the morning of pro-oestrus reproduces the regressive changes that happen in the cyclic corpus luteum (CL) during the transition from pro-oestrus to oestrus, and to analyse the temporal relationships between two characteristic features of structural luteolysis (luteal cell apoptosis and accumulation of macrophages). Cyclic rats (treated at 0900 h with an LHRH antagonist to block LH secretion) were injected at 1000 h with PRL and progesterone and killed at 0, 30, 60, 90 and 180 min after treatment. The number of apoptotic cells increased progressively from 60 min after treatment onward in hormone-treated rats, whereas the number of macrophages did not change throughout the period of time considered. Rats injected with PRL plus progesterone showed significantly greater numbers of apoptotic cells than those injected with PRL alone. The luteolytic effects of progesterone were in keeping with the presence of luteal endothelial cells showing progesterone receptor (PR) immunoreactivity in pro-oestrus. Treatment of rats during dioestrus and pro-oestrus with the specific antioestrogens LY117018 and RU58668 decreased the luteolytic effects of PRL and progesterone and the number of luteal endothelial cells immunostained for PR. These results strongly suggest that the preovulatory PRL surge and the preovulatory increase in progesterone together trigger structural regression of the corpus luteum. This seems to be dependent on oestrogen-driven cyclic changes in PRs in luteal endothelial cells. PMID- 10828857 TI - Possible relationship between changes in islet neogenesis and islet neogenesis associated protein-positive cell mass induced by sucrose administration to normal hamsters. AB - The possible relationship between changes in islet cell mass and in islet neogenesis-associated protein (INGAP)-cell mass induced by sucrose administration to normal hamsters was investigated. Normal hamsters were given sucrose (10% in drinking water) for 5 (S8) or 21 (S24) weeks and compared with control (C) fed hamsters. Serum glucose and insulin levels were measured and quantitative immunocytochemistry of the endocrine pancreas was performed. Serum glucose levels were comparable among the groups, while insulin levels were higher in S hamsters. There was a significant increase in beta-cell mass (P<0.02) and in beta-cell 5 bromo-2'-deoxyuridine index (P<0.01), and a significant decrease in islet volume (P<0.01) only in S8 vs C8 hamsters. Cytokeratin (CK)-labelled cells were detected only in S8 hamsters. INGAP-positive cell mass was significantly larger only in S8 vs C8 hamsters. Endocrine INGAP-positive cells were located at the islet periphery ( approximately 96%), spread within the exocrine pancreas ( approximately 3%), and in ductal cells (<1%) in all groups. INGAP positivity and glucagon co-localization varied according to topographic location and type of treatment. In C8 hamsters, 49.1+/-6. 9% cells were INGAP- and glucagon-positive in the islets, while this percentage decreased by almost half in endocrine extra insular and ductal cells. In S8 animals, co-expression increased in endocrine extra-insular cells to 36.3+/-9.5%, with similar figures in the islets, decreasing to 19.7+/-6.9% in ductal cells. INGAP-positive cells located at the islet periphery also co-expressed CK. In conclusion, a significant increase of INGAP-positive cell mass was only observed at 8 weeks when neogenesis was present, suggesting that this peptide might participate in the control of islet neogenesis. Thus, INGAP could be a potentially useful tool to treat conditions in which there is a decrease in beta-cell mass. PMID- 10828858 TI - Leptin expression in offspring is programmed by nutrition in pregnancy. AB - Birth weight is a determinant of blood leptin concentrations in adults. Since nutrition during pregnancy can affect birth weight, the hypothesis that feed intake during pregnancy alters leptin expression in progeny was examined. Leptin mRNA was measured in subcutaneous adipose tissue and leptin protein was measuredin blood plasma from 59 day old female pigs whose mothers were fed at the same restricted rate except that half were permitted to consume 35% more feed during the second quarter of pregnancy. Leptin mRNA abundance in adipose tissue (P=0.015) and plasma leptin concentration (P=0.01) were higher in progeny from mothers provided with more feed. Body weight at birth was negatively correlated with the abundance of leptin mRNA in subcutaneous fat at 59 days of age (P=0.01). This study shows for the first time that maternal nutrition during pregnancy programs postnatal leptin expression in offspring. PMID- 10828859 TI - High-dose therapy in breast cancer: out of favor but not out of promise. AB - The evidence for use of high-dose chemotherapy and autologous stem cell transplantation for breast cancer still remains inconclusive at best. A number of prospective randomized phase III studies have been either published or presented recently or are underway in North America and Europe. It will be crucial to complete the available prospective randomized phase III study and obtain the data when all studies reach a mature status. Only then will level I evidence become available to determine the efficacy and effectiveness of high-dose chemotherapy and autologous stem cell transplantation in breast cancer. Bone Marrow Transplantation (2000). PMID- 10828860 TI - Allogeneic peripheral blood stem cell transplantation using a fludarabine-based low intensity conditioning regimen for malignant lymphoma. AB - Relapse is a serious complication following high-dose therapy and autologous bone marrow transplantation (ABMT) for malignant lymphoma (ML). Allogeneic transplantation (alloSCT) is a therapeutic option. However, it is associated with a high incidence of transplant-related organ toxicity and mortality. We recently reported fast engraftment and minimal transplant-related toxicity, using fludarabine-based conditioning with reduced amounts of chemotoxic drugs prior to alloSCT. We now present our experience with 23 heavily treated high risk ML patients who underwent matched alloSCT following the same low intensity conditioning. The patients (20 male, three female) were aged 13-63 years. Nineteen had NHL and four HD (resistant disease 12, partial remission 11). Five were post ABMT. Twenty-two patients had fully matched sibling donors, and one a fully matched unrelated donor. Engraftment was fast. There was no rejection or non-engraftment. Organ toxicity was moderate with no liver or renal toxicity >grade II. Four patients developed >grade II graft-versus-host disease (GVHD). Seven patients died - four of grade III-IV GVHD and severe infections, two of bacterial sepsis, one of pulmonary failure. Ten patients are alive after 22.5 (15 37) months. Survival and disease-free survival at 37 months are both 40%. Probability of relapse is 26%. These encouraging results suggest that alloSCT following fludarabine-based low intensity conditioning in high-risk patients merits further evaluation. Bone Marrow Transplantation (2000). PMID- 10828861 TI - The frequency and proliferative potential of megakaryocytic colony-forming cells (Meg-CFC) in cord blood, cytokine-mobilized peripheral blood and bone marrow, and their correlation with total CFC numbers: implications for the quantitation of Meg-CFC to predict platelet engraftment following cord blood transplantation. AB - CFC numbers have shown to correlate with success of engraftment and speed of neutrophil recovery following cord blood (CB) transplantation. To investigate whether the number of Meg-CFC in a CB stem cell preparation might correlate with time to platelet engraftment, we evaluated the frequency of Meg-CFC among all CFC types in 134 CB, 21 adult bone marrow (BM) and 52 cytokine-mobilized peripheral blood (PB) stem cell preparations. The correlation of Meg-CFC with the total number of CFC and mixed cell-CFC was also assessed. The frequency of Meg-CFC was highest in CB and correlated significantly with total CFC numbers (mean 20.8%, correlation coefficient (r) 0.84, P = 0.0001) compared with Meg-CFC from mobilized PB (mean 13.1%, r = 0.29, P = 0.07) and BM (mean 4%, r = 0. 39, P = 0.13). In addition, mixed-cell CFC numbers in CB were highly correlated with the total number of Meg-CFC (r = 0.7, P = 0.0001). No such correlations were found with mobilized PB or BM. We conclude that, based on the high degree of correlation between Meg-CFC and non-Meg-CFC numbers in CB, no additional information concerning time to platelet engraftment would be gained by routinely performing Meg-CFC assays in addition to non-Meg-CFC assays. The fact that CB Meg CFC and mixed-cell CFC are strongly correlated suggests that CB Meg-CFC are more primitive than their counterparts in BM and PB and may reflect the number of stem cells in CB. Bone Marrow Transplantation (2000). PMID- 10828862 TI - G-CSF reduces IFN-gamma and IL-4 production by T cells after allogeneic stimulation by indirectly modulating monocyte function. AB - Despite a 10-fold increase of T cell dose, the incidence and severity of acute GVHD following allogeneic transplantation of G-CSF-mobilized PBSC is not increased compared to BMT. Experimental murine studies demonstrate that G-CSF polarizes donor T cells toward a type 2 cytokine response. To determine whether G CSF alters T cell cytokine responses, we investigated the effects of G-CSF administration on T cell proliferative and cytokine responses to alloantigen and Con A in nonadherent PBMC (NAC) and CD3+ T cells obtained from normal individuals before and after G-CSF administration (10 microg/kg x 4 days). Although T cell proliferative and cytokine (IFN-gamma and IL-4) responses to alloantigen stimulation and Con A were significantly reduced in post-G-CSF NAC, they were restored by the removal of non-T cells from post-G-CSF NAC. Furthermore, there was less T cell alloreactivity in MLR in the presence of autologous post-G-CSF monocytes than in the presence of pre-G-CSF monocytes. This alteration was not replicated in vitro by culturing PBMC with G-CSF. These results suggest that G CSF administration suppresses T cell proliferative and cytokine (IFN-gamma and IL 4) responses to allogeneic stimulation by indirectly modulating monocyte function. Bone Marrow Transplantation (2000). PMID- 10828863 TI - High-dose chemotherapy and stem cell transplantation for patients with stage IV breast cancer without clinically evident disease: correlation of CD34+ selection to clinical outcome. AB - Forty-five patients with metastatic breast cancer without clinically evident disease were treated with thiotepa 750 mg/m2, mitoxantrone 40 mg/m2 and carboplatin 1000 mg/m2 followed by stem cell transplantation to determine the safety and efficacy of CD34+ selection of peripheral blood stem cells. Of these, 15 patients' (group I) stem cells were processed through Baxter Isolex 300 device for CD34+ selection, whereas 30 patients (group II) received unmanipulated stem cells. Toxicity, progression-free survival and survival were compared between these two groups. There was no difference in transfusion requirements, white cell count and platelet recovery and non-hematologic toxicity between the two groups. The survival of patients in group I was 27 months compared to 38 months in group II (P = 0.8). The progression-free survival was 12 months and 13.5 months for group I and group II patients, respectively (P = 0.6). Our results indicate that while there is no adverse effect, there is also no significant advantage of CD34+ selection in terms of progression-free survival and survival in patients with metastatic breast cancer without clinically evident disease. Bone Marrow Transplantation (2000). PMID- 10828864 TI - Cardiac sequelae of doxorubicin and paclitaxel as induction chemotherapy prior to high-dose chemotherapy and peripheral blood progenitor cell transplantation in women with high-risk primary or metastatic breast cancer. AB - Doxorubicin plus paclitaxel has been shown to be an active regimen for metastatic breast cancer and is now frequently used as adjuvant therapy for high-risk primary breast cancer. Initial studies reported a higher than expected rate of cardiac toxicity with this regimen. We studied 105 patients with either high-risk primary breast cancer or metastatic breast cancer who were treated with doxorubicin (60 mg/m2) and 3-h infusions of paclitaxel (175 mg/m2) cycled every 3 weeks. Patients received three cycles of chemotherapy for high-risk primary or four cycles for metastatic disease. Patients then proceeded to high-dose chemotherapy (HDC) (STAMP I cyclophosphamide, cisplatin and carmustine) and peripheral blood progenitor cell transplantation (PBPCT). Patients underwent radionuclide multi-gated angiograms (MUGA) before and following induction chemotherapy and following HDC. During induction chemotherapy 40 (38%) of the patients had a reduction in left ventricular ejection fraction (LVEF). Fourteen had a decrease of 20% or greater and two were mildly symptomatic from CHF. There was additional reduction in the LVEF after HDC with a median value for LVEF of 59% (range, 20-78%). During HDC 10 patients developed clinical signs of congestive heart failure (CHF). Five patients responded to diuretic therapy and did not require any additional treatment. Four patients responded to vasodilation and/or digoxin with improvement in cardiac function. A clinically significant decrease in cardiac function was found in a small number of patients after induction chemotherapy and HDC with PBPCT. The majority of the patients tolerated this regimen without problems. Although there was a decline in LVEF as measured by radionuclide MUGA this did not prevent the majority of patients from proceeding with HDC. Bone Marrow Transplantation (2000). PMID- 10828865 TI - The dismal outcome in patients with acute leukaemia who relapse after an autograft is improved if a second autograft or a matched allograft is performed. Acute Leukaemia Working Party of the European Group for Blood and Marrow Transplantation (EBMT). AB - All patients receiving autografts for acute leukaemia in remission between 1 January 1981 and 31 December 1996 and reported to the European Group for Blood and Marrow Transplantation and had a relapse, were included. The patients underwent an allograft (n = 90, group A), were treated with chemotherapy (n = 2584, group B) or received a second autograft (n = 74, group C). The 2-year survival after relapse was 32 +/- 5%, 11 +/- 1% and 42 +/- 6% in groups A, B and C, respectively. In group A, those with an HLA-A, -B and -DR compatible related or unrelated donor had a 2-year survival of 37 +/- 7% compared to 13 +/- 8% for those receiving a graft from an HLA mismatched donor (n = 20). The following factors were associated with better survival in multivariate analyses: an interval from first autograft to relapse >5 months (P < 0.00001), a first autograft performed later than 1991 (P < 0.00001), patient age below 26 years (median, P < 0.002), group B vs HLA mismatches from group A (P = 0.002), group C vs group B (P < 0.005), patients who were not treated with total body irradiation at first autograft (P < 0.02) and patients in first remission at first autograft (P = 0.02). To conclude, the poor outcome in these patients was improved if a second autograft was feasible (P < 0.005), or if an HLA-matched allograft was performed (NS). Bone Marrow Transplantation (2000). PMID- 10828866 TI - Unrelated bone marrow transplantation in children: outcome and a comparison with sibling donor grafting. AB - The clinical course of 59 children, who underwent BMT during 1988-1998 with a matched unrelated donor (MUD), was compared with 59 case controls receiving a sibling donor marrow. Thirty-eight patients had haematological malignancies while 21 had a nonmalignant disorder. The cumulative incidence of acute GVHD grade II IV was 28% for MUD recipients vs 11% (P = 0.014) for sibling recipients. Extensive chronic GVHD was rare in both groups. The 5-year probability of survival was 52% for MUD vs 77% for sibling recipients (P= 0.014). For children with malignancies the 4-year probability of survival was 52% for MUD vs 67% for sibling recipients with a RFS of 49% vs 62%. In the ALL patients the survival of the MUD recipients was 77% and equalled that of the sibling group. For SAA survival was 43% vs 86% (P = 0.09) and for metabolic disorders 63% vs 89% (P = 0.025). The transplant-related mortality was higher in the MUD group, while death due to relapse was equally distributed. These results of MUD BMT in children compare favourably with most previous reports, and support the use of alternative donors in cases who lack an HLA-identical siblings. Bone Marrow Transplantation (2000). PMID- 10828867 TI - Mycophenolate mofetil (MMF) as therapy for refractory chronic GVHD (cGVHD) in children receiving bone marrow transplantation. AB - Mycophenolate mofetil (MMF) is an alternative immunosuppressant which inhibits the proliferation of T and B lymphocytes. The purpose of the present study was to evaluate the safety and efficacy of MMF as salvage therapy for chronic GVHD (cGVHD) in children receiving allogeneic bone marrow transplantation. Fifteen children, 3-16 years of age, who had received grafts from HLA-compatible siblings (n = 8), partially matched related donors (n= 2) or matched unrelated donors (n = 5), developed extensive cGVHD which had proved unresponsive to standard immunosuppressive therapy. Patients were treated with MMF at the dose of 15-40 mg/kg/day in combination with other immunosuppressive therapy for a median of 4 months (range 1-15 months). The overall response rate (complete or partial response) was 60%. Thirteen percent had only minor responses, whereas 27% of patients had progressive disease. Best responses were seen in patients with GI tract (60% of complete responses) or mouth (33% of complete responses) cGVHD and skin involvement (43% of complete responses) that did not include sclerodermatous manifestations. Once MMF was started, improvements in the clinical manifestations of cGVHD allowed a significant reduction of steroids in 45% of patients and discontinuation in 27% of cases. Six patients (40%) experienced adverse events, with gastrointestinal symptoms predominating. Five patients experienced opportunistic infections. MMF was discontinued after 35-180 days in six patients for the following reasons: parents choice (n = 2), liver toxicity (n = 1), poor compliance (n = 2), and no response (n = 1). In conclusion, these preliminary results suggest that MMF in combination with other immunosuppressive agents may have a role to play in patients with cGVHD. Prospective clinical trials are needed to establish exact indications for therapy and dosage scheduling. Bone Marrow Transplantation (2000). PMID- 10828868 TI - Parotid salivary gland dysfunction in chronic graft-versus-host disease (cGVHD): a longitudinal study in a mouse model. AB - Chronic graft-versus-host disease (cGVHD) is an autoimmune-like phenomenon resulting in morbidity and mortality following allogeneic bone marrow transplantation (BMT). Major salivary gland dysfunction and hyposalivation is one of the prevalent manifestations of cGVHD. We have used the B10.D2 to Balb/C cGVHD mice model in order to assess major salivary gland function in cGVHD, evaluating sialometric, sialochemical and histopathological parameters for almost 3 months. As cGVHD is a chronic debilitating disease it is of vast importance to evaluate these parameters on a prolonged longitudinal basis. We observed significant reduction in parotid salivary flow rate and disturbance in the salivary dynamic function in cGVHD mice in comparison to the normal and syngeneic transplanted controls. On days 18, 25, 46, 56 and 88 the mean flow rates of the cGVHD group were 37.4 +/- 4.4 microl/30 min, 40.5 +/- 4.6 microl/30 min, 32.5 +/- 2.3 microl/30 min, 22.2 +/- 3.2 microl/30 min and 14.8 +/- 3.8 microl/30 min, respectively, values which were lower than those of the syngeneic transplanted controls group by 42% (P < 0.04), 32% (P < 0.03), 44% (P < 0.01), 49% (P < 0.01) and 64% (P < 0.01), respectively. These changes in flow rates were paralleled by changes in the biochemical composition of the saliva. Moreover, the reduction in flow rates correlated with the degree of salivary gland destruction observed in the pathological slides. An inverse correlation was observed between the mean parotid salivary flow rate and the degree of fibrosis observed in the histopathological evaluation of the cGVHD mice (P < 0.01). Maximal flow rate 34.8 +/- 4.6 microl/30 min was observed when no fibrosis was observed while in mice with maximal fibrosis flow rates were minimal. This may point to the pathological mechanism leading to the major salivary gland dysfunction and hyposalivation observed in cGVHD. Thus, it may broaden our knowledge and provide the scientific background for designing better therapeutic strategies for this complication. Bone Marrow Transplantation (2000). PMID- 10828869 TI - Introduction of the oncological pediatric risk of mortality score (O-PRISM) for ICU support following stem cell transplantation in children. AB - Prognostic scoring systems based on physiological parameters have been established in order to predict the outcome of ICU patients. It has been demonstrated that the predictive value of these scores is limited in patients following hematopoietic stem cell transplantation (HSCT). Therefore, we evaluated patients from the Dusseldorf pediatric stem cell transplantation center with regard to predisposing factors and prognostic variables for ICU treatment and outcome. Between January 1989 and December 1998, 180 HSCT have been performed. The clinical, laboratory and HSCT-related parameters such as conditioning treatment, engraftment, GVHD, infections and HSCT toxicity were prospectively recorded and retrospectively analyzed. Established pediatric scoring systems (PRISM, TISS, P-TISS) were applied. Twenty-eight patients required intensive care (16 male, 12 female, median age: 10.9 years (range: 0.4 to 18.9 years), five autologous, 13 allogeneic-related and 10 unrelated transplanted patients). Ventilator-dependent respiratory failure was the most frequent cause of admission to the ICU (n = 23). Fourteen of 28 patients were discharged from ICU, and six of 28 patients achieved a long-term survival (110 to 396 weeks). At admission to the ICU, impaired cardiovascular status, high CRP levels and presence of macroscopic bleeding were each associated with fatal outcome (P < 0.05). The Pediatric Risk of Mortality (PRISM) score was not prognostically significant at the 0.05 level. Long-term survival after discharge from the ICU correlated with HSCT-related parameters such as the type of transplant and severity of GVHD (P = 0.002). By introduction of HSCT related parameters such as severity of GVHD (grade 2: 2 points; grade >2: 4 points), CRP-level (>10 mg/dl: 4 points), and presence of macroscopic bleeding (4 points) into the PRISM score a new oncological PRISM ('O PRISM') score was established. This score significantly correlated with the risk of mortality in the ICU (P = 0.01). In conclusion, the new O-PRISM score accurately characterizes the clinical situation of children requiring ICU treatment following HSCT. It distinguishes more appropriately between success and failure of ICU treatment following HSCT than the standard prognostic scores. It needs to be evaluated in future prospective studies of critically ill children after HSCT. Bone Marrow Transplantation (2000). PMID- 10828870 TI - Growth and endocrine function in children with acute myeloid leukaemia after bone marrow transplantation using busulfan/cyclophosphamide. AB - Longitudinal studies of growth and endocrine function of children with AML transplanted with BUCY are limited. We report a cohort of 23 children with AML transplanted (15 autologous and eight allogeneic) following a single chemotherapy protocol and surviving at least 2 years after BMT. Busulfan was given as a single daily dose. Growth and endocrine function was evaluated yearly from one up to 10 years post transplant (median 4.9 years). The mean height standard deviation score (HtSDS) of the entire group decreased from 0.01 (s.e.m. +/- 0.25) at diagnosis to -0.38 (+/- 0.28) at BMT (P = 0.001). There was no statistically significant difference between HtSDS at BMT and yearly HtSDS from 1 to 5 years post BMT. There was no significant relationship between age at BMT and subsequent change in HtSDS. To date, five of six girls have needed sex steroid replacement. Six of 12 evaluable boys had abnormal gonadotrophins, but none required sex steroid replacement. Children with AML who undergo BMT with BUCY show no significant growth impairment, but gonadal dysfunction is prominent, particularly in girls. Bone Marrow Transplantation (2000). PMID- 10828871 TI - Unrelated umbilical cord blood transplantation in infancy for mucopolysaccharidosis type IIB (Hunter syndrome) complicated by autoimmune hemolytic anemia. AB - This report describes unrelated umbilical cord blood transplantation for a 10 month-old infant boy with mucopolysaccharidosis IIB (Hunter syndrome), an X linked metabolic storage disorder due to deficiency of iduronate sulfatase. Two years after transplant approximately 55% normal plasma enzyme activity has been restored and abnormal urinary excretion of glycosaminoglycans has nearly completely resolved. The boy has exhibited normal growth and development after transplant. Nine months after transplant he developed severe autoimmune hemolytic anemia and required 14 months of corticosteroid treatment to prevent clinically significant anemia. Bone marrow transplantation for Hunter syndrome and post transplant hemolytic anemia are reviewed. Bone Marrow Transplantation (2000). PMID- 10828872 TI - Hematopoietic cell transplantation for mucopolysaccharidosis IIB (Hunter syndrome). AB - Hunter syndrome is an X-linked metabolic storage disorder arising from deficiency of iduronate sulfatase enzyme activity. Despite the successful use of hematopoietic cell transplantation for a variety of lysosomal and peroxisomal storage diseases, limited benefit occurs following transplantation in either the severe or mild forms of Hunter syndrome. A brief ethical commentary is provided on the case of a boy with mucopolysaccharidosis IIB (ie the mild form) who received an unrelated umbilical cord blood transplant to improve his future quality of life. Bone Marrow Transplantation (2000). PMID- 10828873 TI - GVHD dry eyes treated with autologous serum tears. AB - Two cases of GVHD with severe dry eyes are reported where conventional therapy failed to control ocular signs and symptoms. Autologous serum tears, however, resulted in a beneficial clinical effect with marked attenuation of the symptoms. This therapy proved to be safe during 10 months of treatment. Bone Marrow Transplantation (2000). PMID- 10828874 TI - Infusion of lymphocytes obtained from a donor immunised with the paraprotein idiotype as a treatment in a relapsed myeloma. AB - A 48-year-old patient with IgA k multiple myeloma received a BMT from his HLA matched sibling. After transplantation, the disease relapsed. Melphalan therapy followed by reinfusion of haemopoietic blood stem cells collected from the patient led to the improvement of the clinical status, although mixed chimerism and an elevated serum IgA persisted. Successful donor immunisation against an immunogenic preparation of the recipient monoclonal protein was performed before the infusion of donor T lymphocytes (DLI) into the patient. Ten weeks after the lymphocyte infusions, no monoclonal band was evidenced and donor complete chimerism was detected. The patient did not develop GVHD. Once complete remission was achieved, the idiotype vaccine was administered to the patient. Nineteen months after DLI, the patient remains in remission. Bone Marrow Transplantation (2000). PMID- 10828875 TI - An accelerrated-phase CML patient with e19a2 junction of BCR/ABL gene - the first case of transplanted CML with micro bcr. PMID- 10828876 TI - Factor V Leiden mutation as a predisposing factor for veno-occlusive disease following BMT. PMID- 10828877 TI - Toxoplasmosis after hematopoietic stem transplantation. Report of a 5-year survey from the Infectious Diseases Working Party of the European Group for Blood and Marrow Transplantation. PMID- 10828878 TI - Stress signals induce transcriptionally inactive E2F-1 independently of p53 and Rb. AB - One of the common features of cellular response to stress is cell cycle arrest or apoptosis. E2F is one of the key factors which controls cell cycle progression. Overexpression of E2F-1 can also induce apoptosis. In order to understand the role of E2F-1 in cellular response to stress, we studied the E2F-1 response in various cell lines to different types of stress signals including UV irradiation, cisplatin, etoposide and hypoxia. We showed here that the expression level of E2F 1 can be up regulated by the treatment of DNA damage agents as well as hypoxia. The kinetics of E2F-1 increase was dependent on the types of inducer and was similar to that of p53. However, stress signals can induce E2F-1 expression independently of p53 and Rb. Furthermore, the induced E2F-1 was transcriptionally inactive. All these results suggested that E2F-1 may play a very important role in cellular response to stress and this novel role of E2F-1 is independent of its transactivation function. PMID- 10828879 TI - Extinction of rac1 and Cdc42Hs signalling defines a novel p53-dependent apoptotic pathway. AB - Apoptosis is a normal physiological process which eliminates cells that do not receive adequate extracellular signals. One of the pathways signalling apoptosis is controlled by the small GTPases of the Rho family, also involved in cell proliferation, differentiation and motility. Another major apoptosis signalling pathway involves the p53 tumour suppressor which is activated by a variety of stress and mediates growth arrest or apoptosis in normal cells. We show here that upon detachment from the extracellular matrix, fibroblasts undergo rapid apoptosis that can be rescued by constitutive activation of Rac1 and Cdc42Hs GTPases. Conversely, inhibition of Rac1 and Cdc42Hs efficiently triggers apoptosis in adherent cells. Interestingly, apoptosis is not observed in p53-/- cells either cultured in suspension or inhibited for Rac1 and Cdc42Hs activity. Moreover, Rac1 and Cdc42Hs extinction in normal cells activates endogenous p53. Using specific inhibitors of MAPK pathways, we demonstrate that, in our experimental system, p38 signals survival, while ERK activity is required for apoptosis. Our data constitute the first demonstration that Rac1 and Cdc42Hs control pathways that require simultaneous signalling through MAPK ERK and p53 to induce apoptosis. PMID- 10828880 TI - Dominant negative Met reduces tumorigenicity-metastasis and increases tubule formation in mammary cells. AB - Activation of the Met tyrosine kinase growth factor receptor by its ligand HGF/SF has been shown to increase in vitro invasiveness in epithelial cell lines. To study the effect of Met-HGF/SF signaling in breast cancer cells, we transfected met, hgf/sf and dominant negative (DN) forms of met into the poorly differentiated metastatic murine mammary adenocarcinoma cell line DA3. These cells express moderate levels of endogenous Met, which is rapidly phosphorylated in response to HGF/SF treatment. Met+hgf/sf transfection results in significantly increased tumorigenic and metastatic activity in vivo accompanied by reduced tubule formation. DA3 cells transfected with DN forms of Met (DN-DA3) exhibit reduced Met phosphorylation following exposure to HGF/SF. Furthermore, as compared to the parental cells, the DN-DA3 cells exhibit diminished in vitro scattering and invasiveness, while in vivo they display greatly reduced tumorigenicity and spontaneous metastasis. Tumors emanating from DN-DA3 cells injected to BALB/C mice are highly differentiated and display extensive tubule formation. These results suggest that Met-HGF/SF signaling is a determining factor in the delicate balance between differentiation/tubule formation and tumorigenicity-metastasis. Oncogene (2000) 19, 2386 - 2397 PMID- 10828881 TI - High tumoral maspin expression is associated with improved survival of patients with oral squamous cell carcinoma. AB - Maspin, a member of the serpin family of protease inhibitors, is known to have tumor-suppressor functions. However, the association between its expression level and survival has not been demonstrated in human cancer. Using the immunohistochemical technique to examine the expression levels of maspin in 44 cases of oral squamous cell carcinoma (SCC), we found that 66% of the cases expressed low to intermediate levels of maspin and 34% of the cases expressed high levels of maspin. We further examined maspin protein expression in a series of six SCC cell lines from the head and neck, and found that all but one expressed low or no maspin protein. We also compared the clinicopathological features of the oral SCC cases with the maspin expression level, and found that high maspin expression was associated with the absence of lymph node metastasis. More importantly, we showed that higher maspin expression was significantly associated with better rates of overall survival, suggesting that high maspin expression may be a favorable prognostic marker for oral SCC. PMID- 10828882 TI - The impact of bcl-2 expression and bax deficiency on prostate homeostasis in vivo. AB - Prostatic glandular epithelial cells undergo apoptosis in response to androgen deprivation. The molecular determinants of androgen-responsiveness in these cells are incompletely understood. Recent evidence suggests that bcl-2 gene family members may be important in this context. We used the probasin promoter to target a human bcl-2 transgene specifically to the prostate in order to assess its impact on conferring resistance to androgen withdrawal in, otherwise sensitive, prostatic glandular epithelial cells in vivo. We examined the contribution of bax to mediating androgen-responsiveness in prostatic glandular epithelial cells using bax knockout mice. The histologic appearance of the prostates from probasin bcl-2 transgenic mice or bax-/- mice did not differ from those of control littermates. There was no evidence of hyperplastic or neoplastic growth. There was no difference between probasin bcl-2 transgenic mice, bax-/- mice, and control littermates in steady-state levels of apoptosis. Following castration our findings suggest that both bax and bcl-2 may each contribute to the androgen responsiveness of prostatic glandular epithelial cells. It is apparent from these results, however, that bax is not required to mediate cell death in prostatic glandular epithelial cells following castration. A comparison between the apoptotic indices in the ventral prostate from the probasin-bcl-2 and bax-/- mice following castration suggests that the presence of bcl-2 may be a more important indicator of androgen-sensitivity than a deficiency of bax. PMID- 10828883 TI - The chimeric FUS/TLS-CHOP fusion protein specifically induces liposarcomas in transgenic mice. AB - The characteristic t(12;16)(q13;p11) chromosomal translocation, which leads to gene fusion that encodes the FUS-CHOP chimeric protein, is associated with human liposarcomas. The altered expression of FUS-CHOP has been implicated in a characteristic subgroup of human liposarcomas. We have introduced the FUS-CHOP transgene into the mouse genome in which the expression of the transgene is successfully driven by the elongation factor 1alpha (EF1alpha) promoter to all tissues. The consequent overexpression of FUS-CHOP results in most of the symptoms of human liposarcomas, including the presence of lipoblasts with round nuclei, accumulation of intracellular lipid, induction of adipocyte-specific genes and a concordant block in the differentiation program. We have demonstrated that liposarcomas in the FUS-CHOP transgenic mice express high levels of the adipocyte regulatory protein PPARgamma, whereas it is not expressed in embryonic fibroblasts from these animals following induction to differentiation toward the adipocyte lineage, indicating that the in vitro system does not really reflect the in vivo situation and the developmental defect is downstream of PPARgamma expression. No tumors of other tissues were found in these transgenic mice despite widespread activity of the EF1alpha promoter. This establishes FUS-CHOP overexpression as a key determinant of human liposarcomas and provide the first in vivo evidence for a link between a fusion gene created by a chromosomal translocation and a solid tumor. PMID- 10828884 TI - Differential modulation of paclitaxel-mediated apoptosis by p21Waf1 and p27Kip1. AB - The impact of the cyclin dependent kinase (CDK) inhibitors p21Waf1 and p27Kip1 on paclitaxel-mediated cytotoxicity was investigated in RKO human colon adenocarcinoma cells with the ecdysone-inducible expression of p21Waf1 or p27Kip1. Ectopic expression of p27Kip1 arrested cells at G1 phase, whereas p21Waf1 expression arrested cells at G1 and G2. Expression of p21Waf1 after paclitaxel treatment produced much greater resistance to paclitaxel than did expression of p27Kip1. We attributed this difference to the additional block at G2 induced by p21Waf1, which prevented cells from entering M phase and becoming paclitaxel susceptible. Expression of p21Waf1 inhibited p34cdc2 activity and markedly reduced paclitaxel-mediated mitotic arrest, from 87.5 to 23%. In contrast, p27Kip1 expression also inhibited p34cdc2 but reduced mitotic arrest only slightly, from 87. 4 to 74.5%. We concluded that the G2 block produced by p21Waf1, but not by p27Kip1, contributed to their unequal modulation of sensitivity to paclitaxel-mediated apoptosis in RKO cells, and there is no causal relationship between paclitaxel-mediated cytotoxicity and elevation of p34cdc2 activity. PMID- 10828885 TI - CBP/p300 histone acetyl-transferase activity is important for the G1/S transition. AB - Transforming viral proteins such as E1A which force quiescent cells into S phase have two essential cellular target proteins, Rb and CBP/p300. Rb regulates the G1/S transition by controlling the transcription factor E2F. CBP/p300 is a transcriptional co-activator with intrinsic histone acetyl-transferase activity. This activity is regulated in a cell cycle dependent manner and shows a peak at the G1/S transition, suggesting a function for CBP/p300 in this crucial step of the cell cycle. Here, we have artificially modulated CBP/p300 levels in individual cells through microinjection of specific antibodies and expression vectors. We show that CBP/p300 is required for cell proliferation and has an essential function during the G1/S transition. Using the same microinjection system and GFP-reporter vectors, we demonstrate that CBP/p300 is essential for the activity of E2F, a transcription factor that controls the G1/S transition. In addition, our results suggest that CBP HAT activity is required both for the G1/S transition and for E2F activity. Thus CBP/p300 seems to be a versatile protein involved in opposing cellular processes, which raises the question of how its multiple activities are regulated. PMID- 10828886 TI - ETS1 lowers capillary endothelial cell density at confluence and induces the expression of VE-cadherin. AB - Ets1 is a transcription factor expressed in endothelial cells during angiogenesis but its target genes and function in blood vessel formation are still unknown. We have over-expressed Ets1 as a tagged protein in brain capillary endothelial cells and in 3T3 fibroblasts using a retroviral vector. Over-expression of Ets1 reduced by nearly half cell density at confluence of endothelials but not of fibroblasts. As density at confluence is controlled in part by cadherins, this growth arrest could be due to the up-regulation of these cell contact molecules. Indeed, Ets1 increased the expression of the endothelial-specific VE-cadherin without affecting N-cadherin expression levels. In parallel, both a dominant negative mutant of Ets members and an Ets1 anti-sense oligonucleotide inhibited VE cadherin expression in endothelial cells. Ets1 bound to two Ets-binding sites located in the proximal region of the VE-cadherin promoter. Mutation of these sites abolished Ets1-induced transactivation of the promoter. The present work is the first demonstration of a function of Ets1 in the regulation of a specific endothelial marker based on its endogenous gene and protein expression. PMID- 10828887 TI - Cdc25A stability is controlled by the ubiquitin-proteasome pathway during cell cycle progression and terminal differentiation. AB - Members of the cdc25 family are protein phosphatases that play pivotal roles in cell cycle progression. Cdc25A has been shown to be a critical regulator of the G1/S transition of mammalian cells and to be a myc-target gene with oncongenic properties. We investigated the regulation of cdc25A during terminal differentiation using myeloblastic leukemia M1 cells, that can be induced to undergo differentiation into macrophages by interleukin-6 (IL-6) treatment. In this report it is shown that cdc25A protein is degraded by the ubiquitin proteasome machinery in both terminally differentiating and cycling cells. Cdc25A was found to have two major peaks of accumulation during cell cycle progression, one in G1 and the other in S/G2. Evidence was obtained that degradation of cdc25A by the ubiquitin-proteasome machinery in terminally differentiating myeloid cells is accelerated compared to cycling cells. Moreover, deregulated expression of c myc in M1 cells, which had been previously shown to block terminal differentiation, was also found to block IL-6 induced degradation of cdc25A. PMID- 10828888 TI - Breast cancer growth inhibition by delivery of the MDGI-derived peptide P108. AB - Mammary derived growth inhibitor (MDGI) is a member of the family of cytoplasmic fatty acid binding proteins (FABPs), which bind hydrophobic ligands such as fatty acids, retinoids, eicosanoids and prostaglandines. MDGI and an 11 amino acid MDGI derived conserved C-terminal peptide (P108) inhibits growth of normal mammary epithelial cells in tissue and organ culture, but fails to inhibit proliferation of many breast cancer cell lines in vitro. Here, the effects of peptide P108 on tumor growth of MCF-7, MDA-MB468 and MDA-MB231 human breast cancer cell lines in nude mice were tested. To deliver P108 into tumors, a novel peptide production system was applied for expression and secretion of small bioactive peptides in mammalian cells. Functional differentiation was observed in MCF-7 and MDA-MB468 cells upon P108 expression. In addition, EGF-dependent colony formation in soft agar by MDA-MB468 cells was inhibited by secreted P108. Tumor growth in athymic nude mice was suppressed in all three cell lines tested. Furthermore, P108 expressed by MCF-7/P108 cells caused paracrine tumor growth inhibition of MDA MB231 cells. These results indicate that breast cancer inhibition by P108 is independent of binding to hydrophobic ligands and is perhaps mediated by interference with EGF-dependent signaling pathways. PMID- 10828889 TI - Absence of p53-dependent induction of the metastatic suppressor KAI1 gene after DNA damage. AB - The p53 tumor suppressor protein functions to monitor the integrity of the genome. If a damage is detected, p53 binds tightly to specific sequence elements in the DNA and induces the transactivation of genes involved in various growth regulatory processes such as cell cycle progression, DNA repair and apoptosis. A p53-binding site was recently identified in the promoter region of the metastatic suppressor KAI1 gene, suggesting that this gene was a direct transcriptional target of p53. To test the relevance of this hypothesis, we studied the endogenous KAI1 expression in a series of human cell lines with varying p53 status in response to genotoxic treatment as well as in different cellular models exhibiting an inducible p53 activity. Overall, our data indicate that KAI1 expression is not significantly modulated by p53. This observation provides a direct evidence that the presence of a p53-binding site in regulatory domains is not a sufficient criteria to define a p53-transcriptional target gene. PMID- 10828890 TI - Clinical features, anthropometric characteristics, and racial influences on the 'white-coat effect' in a single-centre cohort of 1553 consecutive subjects undergoing routine ambulatory blood pressure monitoring. AB - BACKGROUND: Patients with high office blood pressures but relatively normal readings during daytime ambulatory monitoring have been the subject of much investigation and debate. This clinical finding in part depends on an alerting reaction to the circumstances of the clinical measurement, often described as the 'white-coat effect' (WCE). Little is known of the characteristics of patients that are associated with the white coat effect in a large population of routinely referred patients. OBJECTIVE: To relate the size of the WCE (defined as the difference between office and ambulatory daytime readings) to clinical features that may influence this phenomenon. METHODS: We categorized 1553 consecutive subjects (51.3% men, aged 17-88 years), who had been referred to a single centre for the assessment of suspected hypertension prospectively into three groups: those aged <40, 40-59, and >/=60 years. RESULTS: WCE on systolic blood pressure (SBP) increased significantly with advancing age and was correlated positively to body mass index (BMI), age and treatment. We found significant correlations to sex (higher in women) and race. WCE on diastolic blood pressure (DBP) decreased slightly with advancing age and was correlated positively to BMI and significantly to race and sex. We found no correlation to age or treatment. Caucasians had a greater WCE than did non-Caucasians (P<0. 001 for SBP and DBP) and hypertensives had greater WCE than did normotensives (P<0.0001 for SBP and DBP). Multiple linear regression analysis showed that age and BMI are the most important factors influencing WCE on SBP and DBP. CONCLUSIONS: Factors such as race, age and BMI may exert important influences on the size of WCE possibly via effects on sympathetic nervous system activity. PMID- 10828891 TI - Racial differences in the influence of body size on ambulatory blood pressure in youths. AB - BACKGROUND: African Americans have higher night-time blood pressures than Caucasians do despite their having similar daytime blood pressures. It is well established that body size is related to casual blood pressure. OBJECTIVE: To examine the influence of body size on racial differences in patterns of ambulatory blood pressure. METHODS: Ambulatory blood pressure recordings were performed on 292 healthy children and adolescents (148 African Americans and 144 Caucasian) aged 10-18 years (mean 13+/-2 years). These blood pressures were related to height, weight, body surface area, and body mass index in separate regression models that also included race, sex, and age as factors. RESULTS: The race-by-height interaction was significant for night-time systolic blood pressure (P<0.02), with a significant relationship for African Americans (P<0. 0001), but not for Caucasians. The race-by-weight interaction was significant for night-time systolic blood pressure (P<0.04), also with a greater relationship for African Americans (P<0.0001) than for Caucasians (P<0.03). In addition, the race-by weight interaction was significant for night-time diastolic blood pressure (P<0.04), with a significant relationship for African Americans (P<0.01), but not for Caucasians. Finally, the race-by-body-surface-area interaction was significant for night-time diastolic blood pressure (P<0.05), again with a significant relationship for African Americans (P<0.02) but not for Caucasians. CONCLUSION: Differences in the relationship between body size and blood pressure contribute to the racial differences in patterns of ambulatory blood pressure and should be considered when evaluating patterns of blood pressure in African American youths. PMID- 10828893 TI - A simulation study of the consistency of oscillometric blood pressure measurements with and without artefacts. AB - BACKGROUND: Oscillometric pressure is measured by analysing, in relation to the cuff pressure, low-amplitude cuff-pressure pulsations generated by each arterial pulse. The cuff pressure is sampled at the pulse rate, introducing measurement variations, which are compounded by artefactual pulses. OBJECTIVE: To study the consistency of measurements with and without artefacts using simulated waveforms. METHODS: The Propaq Smartcuf (with and without electrocardiographic synchronization), the Welch Allyn 52 000 (before and after its software had been upgraded), the Critikon DINAMAP 8100 and Compact TS and the Criticare 507 NJC were evaluated. Each monitor recorded 15 determinations at 120/80 (93) mmHg without and with either low-frequency or high-frequency artefacts generated by the Bio-Tek BP-Pump simulator. Consistency of measurements was defined as SD of less than 2 mmHg for at least two of the systolic, diastolic and mean arterial pressures with all less than 3 mmHg. RESULTS: All monitors except the Critikon 8100 satisfied the consistency criteria without artefacts with most SD less than 1 mmHg. Several satisfied the criteria with a severe low-frequency artefact (all recorded SD were less than 6 mmHg). None satisfied the criteria with a severe high-frequency artefact. High systolic blood pressures were typically recorded with a severe tremor artefact, though the Criticare device, which measures during cuff inflation, recorded lower systolic blood pressures. The Propaq device with electrocardiographic synchronization had the lowest variability, with synchronization increasing determination time. CONCLUSION: Oscillometric monitors are more sensitive to a high-frequency artefact than they are to a low-frequency artefact. Signal-processing techniques can improve consistency of measurements. Simulators can evaluate a monitor's consistency with and without artefacts. PMID- 10828892 TI - Electronic activity-monitor-derived sleeping and awake times and diurnal variation of blood pressure. AB - BACKGROUND: Results of a number of studies have indicated that target-organ damage is more pronounced in non-dippers, those in whom the blood pressure falls by less than 10% with the onset of sleep, than it is in dippers with comparable clinic blood pressures. However, the standard use of arbitrarily defined daytime and night-time periods, rather than precise estimates of sleeping time and awake time, could limit the accuracy of estimates of diurnal variation of blood pressure and hence of dipping status. DESIGN AND METHODS: In this study of 102 consecutive patients undergoing ambulatory blood pressure monitoring we compared activity-derived estimates of sleeping and awake blood pressures using electronic activity monitoring and diary records with estimates determined using pre-defined day and night-time periods. The dipping/non-dipping status of each subject was assessed using these three different techniques for defining the awake/asleep time periods. RESULTS: The sleeping/awake times based on the activity monitor, diary and default data were 2356 h+/-55 min/0754 h+/-50 min, 2326 h+/-61 min/0722 h+/-72 min and 2300 h and 0700 h respectively. The percentage systolic/diastolic falls in blood pressure were 18+/-6/18+/-7% with six non-dippers (activity monitor-derived data), 16+/-6/17+/-8% and 12 non-dippers (diary data) and 13+/ 7/15+/-7% and 21 non-dippers (using the pre-set daytime and night-time periods). DISCUSSION: Results of this study demonstrate that the extent of the diurnal variation in blood pressure (and hence dipping status) can differ depending on the technique used to define periods of wakefulness and sleep. PMID- 10828894 TI - Can simulators evaluate systematic differences between oscillometric non-invasive blood-pressure monitors? AB - BACKGROUND: Oscillometric non-invasive blood-pressure (NIBP) monitors estimate the arterial pressure using model-specific signal processing and algorithms. Hence each model's accuracy must be clinically evaluated. Simulators may assist the evaluation, but their ability to do so has not been verified. OBJECTIVE: To investigate whether simulators can detect systematic differences between NIBP monitors. METHODS: We tested whether a simulator can distinguish between the two different algorithms available in a particular monitor, detect calibration errors and detect systematic differences between monitors that are observed clinically. RESULTS: Simulator evaluation correctly detected 1.8 and 4.2 mmHg systolic and diastolic differences between the two Nellcor N-3100 algorithms (with specified 2 and 5 mmHg differences) but found no difference between their mean arterial pressures (as expected from the specification). Simulator evaluations detected calibration adjustments at 80/50, 120/80 and 200/150 mmHg. Simulator and clinical comparisons of two devices of the same type recording slightly different blood pressures were in close agreement, but simulator and clinical comparisons of three different models (Propaq, Critikon DINAMAP and Datex Cardiocap) were not consistently in agreement. The simulators generated oscillometric pulse shapes different from physiological recordings. CONCLUSION: The results suggest that, although simulators can reveal systematic differences between devices of the same model, they cannot be used to detect systematic differences between different NIBP models. This could be at least partly because the oscillometric pulses generated by the simulators are dissimilar to physiologically recorded pulses. PMID- 10828895 TI - Proceedings from the first international consensus conference on self-blood pressure measurement. Forwardp6. PMID- 10828896 TI - Proceedings from a Consensus Conference on Self-Blood Pressure Measurement. Devices and validation. PMID- 10828897 TI - Development of diagnostic thresholds for automated self-measurement of blood pressure in adults. First International Consensus Conference on Blood Pressure Self-Measurement. AB - Blood pressure is usually measured by conventional sphygmomanometry, a procedure fraught with many potential sources of error. Automated techniques of measurement, such as ambulatory monitoring and self-measurement, reduce the limitations of conventional sphygmomanometry. However, the diagnostic thresholds applicable for conventional sphygmomanometry cannot be extrapolated to automated measurements. During the past 10 years criteria for normality have gradually been developed for ambulatory blood pressure monitoring. First, the distributions of the ambulatory blood pressure in normotensive subjects and in untreated hypertensive patients who had initially been classified on the basis of their conventional blood pressure were studied. Second, epidemiological studies were performed to investigate the distribution of the conventional and ambulatory blood pressures in the population at large. Third, authors of several studies have now validated the preliminary thresholds for ambulatory monitoring against left ventricular hypertrophy, other signs of target-organ damage and the incidence of cardiovascular complications. Finally, authors of clinical trials investigated whether it is beneficial to patients and cost-effective to diagnose and treat hypertension on the basis of ambulatory monitoring rather than under the sole guidance of conventional sphygmomanometry. For systolic/diastolic measurements, the upper limits of normotension include 130/80, 135/85 and 120/70 mmHg for the 24 h, daytime and night-time blood pressures, respectively. Whereas for ambulatory monitoring a large body of evidence currently supports the proposed diagnostic thresholds, for the self-recorded blood pressure, to a large extent, this evidence must still be collected. In pursuing this goal, the methods applied for ambulatory monitoring may serve as a template. On the basis of a meta analysis of summary statistics of published articles and a meta-analysis of data from individual subjects, 135/85 mmHg is likely to be the upper limit of normality for the self-measured blood pressure. Obviously, this threshold is preliminary and must be further validated in prognostic studies. However, the present proposal could guide clinicians who wish to use self-measurement to refine the diagnosis and the management of hypertension based on conventional sphygmomanometry. PMID- 10828898 TI - User procedure for self-measurement of blood pressure. First International Consensus Conference on Self Blood Pressure Measurement. AB - OBJECTIVE: To review the medical literature regarding the methodology of self measurement of blood pressure and to provide some practical recommendations regarding protocol and procedure for measurement, documentation and analysis of data, choice and provision of devices and education of patients and physicians. PROCEDURE AND PROTOCOL FOR MEASUREMENTS: Technical recommendations regarding measurement of blood pressure do not differ from usual guidelines. Frequency of measurement remains a matter of discussion. At the beginning of the self measurements of blood pressure and during the titration phase, there should be a 7-day measurement period with two measurement of blood pressure each morning and two measurements in the evening at pre-stipulated times. For long-term observation, a minimum measurement period of 1 week per quarter is required. The minimum number of measurements performed during each period should be a total of 12 readings recorded within seven working days. Depending on individual needs (e.g. poor compliance) or for pharmacologic studies, a higher frequency of readings could be used. DOCUMENTATION AND ANALYSIS OF DATA: Owing to the lack of reliability of patients' diaries, the use of printer-equipped or memory-equipped devices is advocated. All the recorded data, except those obtained on the first day, must be used for analysis. CHOICE AND PROVISION OF DEVICES: Self-measurement of blood pressure should be performed with validated fully automated devices using a brachial cuff. The preference should be given to apparatus offering the possibility of storing and transmitting measurements. Wrist apparatus should be used with caution due to the risk of measurement errors if it is used inappropriately. A manual device should be considered for patients suffering from irregular cardiac rhythms and patients with large or small arm circumferences, since automated devices have not been validated for use in these situations. Reimbursement of hypertensive patients using validated devices should be considered, so long as they are adequately trained and supervised. EDUCATION OF PATIENTS: In a therapeutic perspective, self-measurement of blood pressure should be performed by trained patients under the supervision of their practitioner. Teaching must be performed by skilled staff in hypertension centers and ultimately in general practice. Self-measurement of blood pressure is to be recommended for any hypertensive patient who is sufficiently motivated to participate in the treatment of his own hypertension. Patients with physical problems or mental disabilities that make them unable to perform or to understand the measuring technique represent the limits of the method. Education of patients must encompass information about hypertension and cardiovascular risk, blood pressure-measurement procedures, advice on items of equipment and their proper use, protocols, and interpretation of data. A patient's proficiency must be checked before he or she should be considered competent at performing the procedure. Annual reevaluation is required. PMID- 10828899 TI - Consensus Conference on Self-blood pressure measurement. Clinical applications and diagnosis. AB - The present study was aimed at reviewing the medical literature devoted to the clinical applications of self-blood pressure monitoring (SBPM) and at providing some recommendations regarding the use of SBPM for diagnostic purposes. The lack of reliability of conventional blood pressure (BP) measurement is largely related to the extreme variability of BP over time. SBPM provides a large number of readings and can be used to predict the results of repeated clinical measurements. The use of SBPM in the diagnosis of white coat hypertension can be proposed as a screening test: if it gives a positive result (a low home BP), it should be confirmed by ambulatory BP monitoring (ABPM). SBPM could improve patients' compliance with medication. Last, SBPM may be cost-effective for the management of hypertensive patients, by reducing costs of medication, number of clinic visits and costs of cardiovascular morbidity. Compared with ABPM, SBPM seems to have a less value for the initial diagnosis of hypertension and for predicting prognosis. In contrast, it should be of more value for the long term follow-up of patients with white coat hypertension and for the evaluation of treatment efficacy in patients with sustained hypertension. The use of SBPM in diabetic hypertensives, in pregnant women and in the elderly is encouraged, but needs further evaluation. PMID- 10828900 TI - Prognostic significance of self-measurements of blood pressure. AB - BACKGROUND: Self-measurements of blood pressure may offer some advantage in diagnostic and therapeutic evaluation and in management of patients. However, the most important limitation of self-measurement is that there are limited data available about the prognostic value of this information. RESULTS: Authors of several previous reports demonstrated that self-measurement reflects target-organ damage better than does casual measurement of blood pressure. So far, investigators in Tecumseh and Ohasama studies have provided pilot data on prognostic value of self-measurements. Investigators in Ohasama study demonstrated that self-measurements predict cardiovascular morbidity and mortality and all-cause mortality better than do casual measurements of blood pressure. Investigators in Tecumseh study demonstrated that self-measurement can predict future development of sustained hypertension and of diastolic dysfunction. These preliminary results suggest that self-measurements have strong predictive power for endpoints and surrogate measures of cardiovascular target organ damage. CONCLUSION: The final answer on the prognostic significance of self measurement has not been given. Prognostic studies designed to compare casual measurement of blood pressure, self-measurement, and ambulatory blood pressure monitoring are needed. PMID- 10828901 TI - Self-measurement of blood pressure in clinical trials and therapeutic applications. AB - Self-measurement of blood pressure (SMBP) is increasingly used to assess blood pressure outside the medical setting. A prerequisite for the wide use of SMBP is the availability of validated devices providing reliable readings when they are handled by patients. This is the case today with a number of fully automated oscillometric apparatuses. A major advantage of SMBP is the great number of readings, which is linked with high reproducibility. Given these advantages, one of the major indications for SMBP is the need for evaluation of antihypertensive treatment, either for individual patients in everyday practice or in clinical trials intended to characterize the effects of blood-pressure-lowering medications. In fact, SMBP is particularly helpful for evaluating resistant hypertension and detecting white-coat effect in patients exhibiting high office blood pressure under antihypertensive therapy. SMBP might also motivate the patient and improve his or her adherence to long-term treatment. Moreover, SMBP can be used as a sensitive technique for evaluating the effect of antihypertensive drugs in clinical trials; it increases the power of comparative trials, allowing one to study fewer patients or to detect smaller differences in blood pressure than would be possible with the office measurement. Therefore, SMBP can be regarded as a valuable technique for the follow-up of treated patients as well as for the assessment of antihypertensive drugs in clinical trials. PMID- 10828902 TI - Inflammatory bowel disease up to 1932. AB - Inflammatory bowel diseases have been a major interest of generations of Mount Sinai Hospital gastroenterologists. Although clinical descriptions of diarrhea with or without blood go back thousands of years, clear distinctions between enteritis and ulcerative colitis were possible only in the 19th century. At that time, many case reports were published of, in retrospect, classical regional enteritis. The term "ulcerative colitis" dates from 1888; the introduction of the electric sigmoidoscope soon after made it possible to make proper diagnosis of ulcerative colitis and distinguish it from infective dysentery, membranous mucous or catarrhal colitis, and nervous diarrhea. Doctors at The Mount Sinai Hospital adopted this diagnostic approach in the 1870s and 1880s, and were particularly interested in patients with tuberculosis-like ileocecal disease without tubercle bacilli. Articles were written by Weiner in 1914, Moschcowitz and Wilensky in 1923 and 1927, and Goldfarb and Suissman in 1931. Dr. A.A. Berg, in 1925, encouraged his assistant Leon Ginzburg to conduct a study of the inflammatory granulomatous diseases of the bowel, when Ginzburg and Gordon Oppenheimer were working in Dr. Paul Klemperer's laboratory. Initial reports came in 1927 and 1928, but Ginzburg and Oppenheimer "in conjunction with Dr. Burrill B. Crohn" presented a definitive paper, "Non-specific Granulomata of the Intestine," on May 2, 1932, to the American Gastro-Enterological Association. On May 13, 1932, Dr. Crohn presented a paper on "Terminal Ileitis" to the American Medical Association; this was published later that year with the title "Regional Ileitis: A Pathologic and Chronic Entity," under the authorship of Crohn, Ginzburg and Oppenheimer. PMID- 10828903 TI - Inflammatory bowel disease after 1932. AB - The clinical diseases of ulcerative colitis (UC) and Crohn's disease (CD) were defined by 1932-1933. After that, the major conceptual developments were the recognition that regional enteritis could clearly involve the colon, and that cancer and toxic megacolon could occur in both CD and UC. In the last half of the 20th century the main thrust of gastroenterology at The Mount Sinai Hospital has been in inflammatory bowel disease (IBD), with contributions to extra-intestinal manifestations, measurement of clinical activity in CD, the natural history of the placebo arm of controlled trials, complications and therapy with corticosteroids, 5-ASA, 6-mercaptopurine, immunomodulators and cyclosporine. Actuarial life tables were introduced for postoperative recurrence and re operation rates, as well as for quality of life analysis. Two forms of CD were defined, perforating and non-perforating, and the role of the fecal stream was explored in light of the higher risk of recurrence after operations with anastomosis as compared with ileocolostomy. PMID- 10828904 TI - The history of surgery for Crohn's disease at The Mount Sinai Hospital. AB - Almost certainly, the physicians and surgeons of The Mount Sinai Hospital cared for patients with inflammatory bowel disease prior to 1932. However, the accepted beginning of the surgery of granulomatous inflammatory bowel disease (IBD) and Crohn's disease (CD) at our institution occurred when the landmark paper by Crohn, Ginzburg, and Oppenheimer was published in 1932. As a major referral center for patients with both medical and surgical complications of IBD, the surgical service has had a long and abiding interest in the disease. This review highlights the major contributions of our staff to the management of this illness over the past 67 years. Despite major innovations in both medical and surgical management of patients with Crohn's disease, individuals suffering from this condition are ideally managed by a multidisciplinary team. PMID- 10828905 TI - Pioneer gastroenterological radiologic studies. AB - In the middle third of the 20th century, Mount Sinai radiologists were able to describe and establish specific radiologic criteria for the diagnosis of many gastrointestinal diseases. They then delineated specific radiologic patterns to diagnose such diverse conditions as inflammatory bowel disease of the small bowel and colon, protein-losing disorders, vascular disease of the small bowel, benign and malignant tumors, metastases and lymphoma of the gastrointestinal tract. Richard H. Marshak, Bernard S. Wolf, and Mansho T. Khilnani were the leaders in these radiologic investigations which established criteria that enabled generations of subsequent radiologists to arrive at definitive diagnoses. PMID- 10828906 TI - IBD: immunologic research at The Mount Sinai Hospital. AB - An evolution in our understanding of the inflammatory bowel diseases (IBD), ulcerative colitis and Crohn's disease, correlates with increased knowledge of the function of the mucosal immune system. In the early 1960s and 1970s, IBD was considered to be an autoimmune disease in which there was a directed attack by humoral and cellular elements of the immune system against intestinal tissues. These studies did not withstand the test of time, and from the 1970s through the 1990s there was a growing appreciation that defects in cellular immunity, not auto-reactive in nature, played a larger role in disease pathogenesis. Research at Mount Sinai focused in on these cellular T cell defects and helped pave the way for the current model of disease pathogenesis. PMID- 10828907 TI - A history of immunosuppressive drugs in the treatment of inflammatory bowel disease: origins at The Mount Sinai Hospital. AB - Much of what we know about the role of immunopathologic mechanisms in causing Crohn's disease and ulcerative colitis originated from research at The Mount Sinai Hospital. The authors were privileged to have been able to share in this undertaking, along with many others, including Moschcowitz, Klemperer, Otani, Crohn, Ginzburg, Oppenheimer, Garlock, Lyons, Marshak, Janowitz, Aufses, Waye, Greenstein, Sachar, Meyers, Gelernt, Mayer, Lichtiger and Kornbluth. In medical history, elucidation of disease processes is often serendipitous. Transplant surgery was successful because of the discovery by Hitchings and Elion of 6 mercaptopurine (6-MP) and azathioprine, which inhibited rejection. And the concept of immunosuppression slowly evolved into possible treatment of any disease thought to be caused by autoimmunity. This includes those diseases of the bowel seen so frequently at The Mount Sinai Hospital: ileitis, granulomatous colitis, ileocolitis, and ulcerative colitis. This paper depicts the progressive role of immunosuppressive drugs, from corticosteroids to 6-mercaptopurine, cyclosporine and anti-tumor necrosis factor, in both the treatment and understanding of the pathogenesis of Crohn's disease and ulcerative colitis. Major contributions to these treatments have come from physicians and surgeons with roots at The Mount Sinai Hospital. PMID- 10828908 TI - Cancer in inflammatory bowel disease. AB - The first case of cancer in inflammatory bowel disease (IBD) was reported at The Mount Sinai Hospital in 1925 in a patient with ulcerative colitis (UC). In 1956, carcinoma of the jejunum was described in a patient with regional enteritis (Crohn's disease [CD]). IBD cancers are preceded by dysplasia, and the relative risk increases with duration of the IBD. CD cancers are more proximally distributed than are UC cancers. Both tend to occur at the site of the overt disease and both develop at earlier ages (47 UC, 50 CD) than in the de novo colorectal cancer (70 years). The absolute cumulative colon cancer frequencies (8% UC, 7% CD) are identical after 20 years, emphasizing the importance of regular surveillance in both types of IBD. Moreover, the increased risk of colon cancer exists in patients with CD even when CD is confined to the small bowel, and patients with IBD have increased risks of developing extraintestinal and reticuloendothelial tumors in both CD and UC, as well as ano-vulval and malignant melanoma in CD. Colitic colorectal cancers are often diffuse, extensive, multiple and right-sided with insidious presentation. The prognosis is no worse after operation than that of de novo colon cancer. Most small bowel cancers in CD are adenocarcinomas, rather than sarcomas, and present at a younger age, more diffusely and more distally than de novo cancers, usually making them undiagnosable at a curable early stage; indeed, two-thirds present with intestinal obstruction. Strictures of the colon are common in patients with IBD, and they have a 10-fold risk for colon cancer, 30-fold for UC, and 6-fold for CD. The risk increases with disease duration. The indications for surgery are absolute, relative and incidental, and the procedures include segmental resection, total proctocolectomy, subtotal colectomy and palliative procedures. PMID- 10828909 TI - The small intestine. AB - Clinical investigation of the small bowel at The Mount Sinai Hospital began with David Adlersberg's arrival in 1931. His research interests were in bile acids, cholesterol, carotene, and vitamin A. In 1952, he was given a Nutrition Laboratory and later, a Nutrition Clinic. His vitamin A tolerance test and interest in malabsorption led him to a comprehensive study of sprue, the separation of the tropical and non-tropical forms, and their different etiologies and treatments. Adlersberg's work was complemented by (a) Marshak and Wolf's radiologic examination of the small bowel (especially in sprue and other malabsorption disorders); (b) Gerson s perfusion experiments; and (c) Friedman, Waye and Wolf's motility studies. Lieber and his colleagues explored the deleterious effects of alcohol on the function and structure of the small intestine. Gerson explored the nutrition of patients with Crohn's disease of the small intestine, especially after extensive resection or bypass leading to ascorbic and folic acid deficiencies and hypergastrinemia. PMID- 10828910 TI - Non-specific granulomata of the intestine (inflammatory tumors and strictures of bowel). 1932. PMID- 10828911 TI - Regional ileitis: a pathologic and clinical entity. 1932. PMID- 10828912 TI - Pin loosening in a halo-vest orthosis: a biomechanical study. AB - STUDY DESIGN: The cranial pin force history of a halo-vest orthosis was measured using an instrumented halo in a clinical study with three patients. Pin force values at the time of halo-vest application and at subsequent clinical visits during the halo-vest wear period were compared. OBJECTIVES: To document the pin force reduction in the cranial pins of a halo-vest orthosis in vivo. SUMMARY OF BACKGROUND DATA: The halo-vest is an orthosis commonly used to immobilize and protect the cervical spine. An important problem with halo-vest use is pin loosening. There have been no previous reports of pin force history in vivo. METHODS: A custom-built strain-gauged, open-ring halo was used to measure the compressive force and superiorly-inferiorly directed shear forces produced at the tips of the two posterior pins. The instrumented halo was applied to three patients with cervical spine fractures. Pin force measurements were recorded at the time of halo application and at subsequent follow-up visits during the entire treatment period. RESULTS: A mean compressive force of 343 +/- 64.6 N was produced at the pin tips during halo application with the patient in a supine position. On average, the compressive forces decreased by 83% (P = 0.002) during the typical halo-vest wear period. The compressive forces were substantially greater than the shear forces, which averaged only -11+/-30.2 N at the time of halo application and which did not change significantly with time. CONCLUSIONS: The study confirmed the hypothesized decrease in the compressive pin forces with time. All patients had developed at least some clinical symptoms of pin loosening at the time of halo-vest removal. PMID- 10828913 TI - Delayed apnea in patients with mid- to lower cervical spinal cord injury. AB - STUDY DESIGN: A retrospective study of 36 patients with mid- to lower cervical spinal cord injury (CSCI) divided into two groups based on whether delayed apnea developed. OBJECTIVES: To determine nonpulmonary risk factors associated with the development of delayed apnea in mid- to lower cervical spinal cord injury. SUMMARY OF BACKGROUND DATA: Patients with mid- to lower cervical spinal cord injury are generally at lower risk of developing respiratory failure than those with high cervical spinal cord injury. Respiratory failure manifesting as sudden apnea may occur days or even weeks after injury without any pulmonary complications in such patients. METHODS: An index group of eight patients with complete mid- to lower cervical spinal cord injury in whom delayed catastrophic apnea occurred were reviewed. Another group of 28 patients with cervical spinal cord injury of identical magnitude and presentation but without respiratory failure served as the control group. Six parameters presumed to be related to the delayed apnea were analyzed. RESULTS: The extent of cord lesions was significantly different, being diffuse in most of the index patients, but focal in the majority of the control patients (P<0.001). Involvement of the C4 segment of cord appeared to be more frequent in the index group; however, the difference was not statistically significant (P = 0.091). The incidence of transient bradycardia (P<0.01) and dyspnea (P<0.001) in the index group was significantly higher than in the control group. Paralytic ileus was a much rarer event and found to be unrelated to the occurrence of apnea. In five of the eight index patients, the apnea occurred during sleep. Six of the eight index patients died of it. CONCLUSIONS: Delayed but devastating apnea may develop in patients with mid- to lower cervical cervical spinal cord injury, even when they are clinically stable and free from any pulmonary complications. The presence of diffuse, extensive cord lesions, respiratory distress, or bradycardia with or without associated hypotension, however transient and self-limited, should be regarded as warning signs. Sleep was found to be a risky period of time. PMID- 10828914 TI - Nonoperative management of dens fracture nonunion in elderly patients without myelopathy. AB - STUDY DESIGN: A retrospective review of elderly patients treated without surgery for chronic mobile nonunions of the odontoid process. Patients were observed on an annual basis with clinical examinations and flexion/extensions plain film radiographs. OBJECTIVES: To evaluate the clinical and radiographic results of elderly patients without myelopathy treated without surgery for dens fracture nonunion. SUMMARY OF BACKGROUND DATA: Because of the risk of progressive myelopathy or sudden neurologic injury, many surgeons recommend operative stabilization for patients with mobile dens nonunions who are able to withstand an operation. There is, however, a lack of information about the radiographic and neurologic progression of dens nonunions. Although a less aggressive surgical approach has been recommended by some authors for elderly or medically compromised patients with acute fractures, long-term follow-up evaluation of patients with resulting nonunions has not been reported. METHODS: A series of elderly patients with chronic, unstable, dens nonunions without myelopathy were treated with a nonoperative treatment protocol. Patients were informed of the nature of their lesion, including the risk of acute or chronic spinal cord injury and the options for operative treatment. Patients were evaluated yearly for clinical and radiographic progression. No intervention to slow progression of atlantoaxial instability was undertaken. RESULTS: None of the patients developed myelopathic symptoms during the follow-up period, and no patient experienced more than a 1 mm radiographic increase in atlantoaxial excursion. None of the reported patients had less than 14 mm available for the spinal cord in either flexion or extension at the start of clinical monitoring. CONCLUSIONS: Although further follow-up evaluation is needed, the authors believe on the basis of this review that this treatment protocol may be considered for patients who are poor candidates for surgical fusion. PMID- 10828915 TI - Stability of myelomeningocele spines treated with the mayfield two-stage anterior and posterior fusion technique. AB - STUDY DESIGN: Retrospective review of the results of operative treatment for paralytic thoracolumbar lordoscoliosis in six consecutive myelomeningocele patients treated with the Mayfield two-stage combined anterior and posterior fusion and instrumentation technique. OBJECTIVES: To assess the long-term stability of myelomeningocele spines treated with the Mayfield technique. SUMMARY OF BACKGROUND DATA: An initial description of the technique by Mayfield and an early study of the evolution of the two-stage combined anterior and posterior instrumented fusion of paralytic lordoscoliosis in myelomeningocele spines are available. METHODS: Study of the six individual patients showed that the mean age at the time of surgery was 13 years and 8 months (range, 9 years 9 months to 15 years 10 months). At the time of surgery, the major lumbosacral curve averaged 81 degrees (range, 52-137 degrees ), pelvic obliquity was 28 degrees (range, 1-48 degrees ), torso decompensation was 2.8 cm (range, 0-6.0 cm), thoracic kyphosis was 60 degrees (range, 25-93 degrees ), lumbar lordosis was 106 degrees (range, 55-151 degrees ), and sacral anteflexion was 78 degrees (range, 22-117 degrees ). The mean duration of follow-up evaluation was 13 years 5 months (range, 8 years 10 months to 16 years 11 months). RESULTS: Clinical and radiographic evaluation showed that immediately after surgery, the lumbosacral curve was corrected to an average of 25 degrees and at last follow-up evaluation was 25 degrees, for a final correction of 69%. Pelvic obliquity was corrected to 7 degrees, 5 degrees at last follow-up evaluation, for a 82% correction. Torso decompensation was corrected to 1.4 cm initially and finally to 0.3 cm, for an 89% correction. Kyphosis was corrected to 52 degrees, finally to 50 degrees (17% decrease); lordosis to 73 degrees and finally to 67 degrees (37 degrees decrease); and sacral anteflexion corrected to 51 degrees and finally to 56 degrees (28% decrease). In one patient, the tip of the longest rod displaced from the uppermost hook and was replaced with maintenance of correction. Another patient had dehiscence of the pelvic portion of the posterior wound, with secondary bacterial contamination, and healed by secondary intention. CONCLUSIONS: The Mayfield technique effectively corrected and stabilized these difficult myelomeningocele spinal deformities, using distraction against square-holed hooks seated on the sacral alae, which contain the best-quality bone in these hypoplastic pelves. PMID- 10828916 TI - Longitudinal changes in trunkal balance after selective fusion of King II curves in adolescent idiopathic scoliosis. AB - STUDY DESIGN: A retrospective study was performed on the longitudinal changes of the trunkal balance in King II curves treated with selective posterior fusion of the thoracic curve. OBJECTIVES: To determine the effect of selective fusion on the coronal and sagittal plane balance in King II adolescent idiopathic scoliosis by analyzing the changes in shoulder level, pelvic tilt, trunk shift, centering of fusion mass, changes in the T11-L1 sagittal angle, and behavior of the unfused lumbar curve and its correlation with the end level of fusion. SUMMARY OF BACKGROUND DATA: It has been shown that selective fusion of the thoracic curve in a King II curve is associated with good results and arrest of lumbar curve progression in selected cases. Detailed quantitative analysis of the longitudinal changes and correlation between various clinical and radiologic parameters was not readily available in the literature. METHODS: This study investigated 24 patients with King II adolescent idiopathic scoliosis treated with Harrington rod and segmental spinous processes wiring in a 10-year period with follow-up periods of 3 to 8 years. Clinical and radiologic parameters were analyzed longitudinally during the preoperative and immediate postoperative period, then at 6 months, 1 year, 3 years, and final follow-up assessment. RESULTS: Progressive improvement in the trunk shift to within 2 cm of the center sacral line together with progressive leveling and stabilization of the shoulder and pelvic tilt was noted during the first year after surgery. Gradual movement of the Harrington rod toward the center sacral line assuming a "straight rod sign" with a rod to center line distance of less than 1 cm was found in 90% of the cases. Improvement of the sagittal alignment with no significant residual junctional kyphosis also was found. The unfused lumbar curve improved in both the coronal and sagittal plane and did not show any further progression. Patients whose lower end level of fusion was at T12 had a better percentage of lumbar curve correction than those that ended at L1. CONCLUSIONS: Selective thoracic fusion for King II idiopathic scoliosis curve can achieve acceptable coronal and sagittal plane balance of the spine. The rod to centersacral line distance is a helpful parameter in assessing the results and prognosis of surgically treated patients. PMID- 10828917 TI - Idiopathic scoliosis: the relation between the vertebral canal and the vertebral bodies. AB - STUDY DESIGN: The axial length of the vertebral canal and the anterior aspect of the vertebrae were measured in 36 skeletons, 15 with probable idiopathic scoliosis. OBJECTIVES: To compare the discrepancy in length of the vertebral canal and the anterior spinal column in skeletons having probable idiopathic scoliosis with the degree of deformity. SUMMARY AND BACKGROUND DATA: In idiopathic scoliosis, the vertebral bodies rotate toward the convexity of the curve, whereas the vertebral canal tends to retain a midline position. The vertebral canal therefore will be relatively short. The degree of shortening has not been described previously, nor its relation with the degree of deformity. METHODS: The axial length of the vertebral canal and the anterior aspect of the vertebral bodies were measured in 36 skeletons: 8 with normal spines, 13 with kyphosis, and 15 with probable idiopathic scoliosis. The relative shortening in the scoliotic spines was correlated with the Cobb angle and the degree of rotation. RESULTS: No significant difference in length was found between the vertebral canal and the vertebral column in the normal spines. The kyphotic spines had canals significantly longer than the vertebral length (P<0.025). All but one of the scoliotic spines had short vertebral canals (P<0.01). The degree of discrepancy was related to the Cobb angle (r = -0.50; P< 0.05), and particularly to the degree of rotation (r = -0.88; P< 0.001). CONCLUSIONS: The findings have surgical and etiologic implications. The results are consistent with a conceivable hypothesis that in some patients with idiopathic scoliosis, there may be impaired growth in the length of the spinal cord, the posterior elements are tethered, and as the vertebral bodies continue to grow, they become lordotic and then rotate. PMID- 10828918 TI - Scoliosis in rhythmic gymnasts. AB - STUDY DESIGN: An anamnestic, clinical, radiographic study of 100 girls actively engaged in rhythmic gymnastics was performed in an attempt to explain the higher incidence and the specific features of scoliosis in rhythmic gymnastic trainees. OBJECTIVES: To analyze the anthropometry, the regimen of motion and dieting, the specificity of training in rhythmic gymnastics, and the growth and maturing of the trainees, and to outline the characteristics of the scoliotic curves observed. An etiologic hypothesis for this specific subgroup of scoliosis is proposed. SUMMARY OF BACKGROUND DATA: The etiology of scoliosis remains unknown in most cases despite extensive research. In the current classifications, no separate type of sports-associated scoliosis is suggested. METHODS: The examinations included anamnesis, weight and height measurements, growth and maturing data, eating regimen, general and back status, duration, intensity, and specific elements of rhythmic gymnastic training. Radiographs were taken in all the patients with suspected scoliosis. The results obtained were compared with the parameters of normal girls not involved in sports. RESULTS: A 10-fold higher incidence of scoliosis was found in rhythmic gymnastic trainees (12%) than in their normal coevals (1.1%). Delay in menarche and generalized joint laxity are common in rhythmic gymnastic trainees. The authors observed a significant physical loading with the persistently repeated asymmetric stress on the growing spine associated with the nature of rhythmic gymnastics. Some specific features of scoliosis related to rhythmic gymnastics were found also. CONCLUSIONS: This study identified a separate scoliotic entity associated with rhythmic gymnastics. The results strongly suggest the important etiologic role of a "dangerous triad": generalized joint laxity, delayed maturity, and asymmetric spinal loading. PMID- 10828919 TI - The rates of false-positive lumbar discography in select patients without low back symptoms. AB - STUDY DESIGN: Experimental disc injections in subjects with no history of low back symptoms. OBJECTIVE: To determine in an experimental setting the relative pain response and pain-related behavior in selected subjects without a history of low back pain undergoing lumbar discography. This study aimed to select a study population that more closely represented patients undergoing discography in clinical practice. SUMMARY OF BACKGROUND DATA: Previous work has shown that in young, healthy men with little degenerative disc disease and no history of low back problems, discographic injections usually did not cause significant pain. This group differed from the patients who usually undergo discographic evaluation in clinical practice. Most clinical patients are older, have significant degenerative disc disease, have behavioral changes associated with chronic pain, and often have confounding psychosocial troubles. The authors undertook to study discography in subjects without low back pain but with clinical profiles similar to patients undergoing discography in clinical practice. METHODS: Twenty-six individuals, mean age 43 years, with no history of low back pain had lumber discography according to the strict protocol of Walsh et al. Of these, 10 were pain-free; 10 had chronic neck and arm pain, but no low back symptoms; and 6 had primary somatization disorders without low back symptoms.- RESULTS: Significant positive pain response and pain-related behavior with discography were found in 10% of the pain-free group, in 40% of the chronic cervical pain group, and in 83% of the somatization disorder group completing the injections. Twenty-four subjects had negative control discs. Discs with annular disruption were more likely to be painful on injection, particularly in those individuals with ongoing compensation issues, chronic pain, or abnormal psychological testing.- CONCLUSION: If strict criteria are applied, the rate of false-positive discography may be low in subjects with normal psychometric profiles and without chronic pain. Significantly painful injections were very common in subjects with annular disruption and chronic pain or abnormal psychometric testing. PMID- 10828920 TI - Cervical discography: clinical implications from 12 years of experience. AB - STUDY DESIGN: Positive pain responses provoked in an inclusive series of cervical discograms performed over a 12-year period were categorized by level and reviewed. OBJECTIVES: To report the prevalence of cervical pathology over an entire series of patients, to determine whether a reproducible pattern of concordant pain could be associated with each symptomatic level identified, and to calculate the rate of complications. SUMMARY OF BACKGROUND DATA: Cloward wrote the first articles explaining the technique of cervical discography and reported on the pain responses induced. Currently, the technique is viewed as an invaluable diagnostic tool, but it also is criticized for failing to contribute unique information beyond that available from imaging studies despite the inherent risks. METHODS: A series of 173 cervical discograms performed over 12 years was examined. Pain responses provoked and recorded during discography were grouped by disc level and examined for recurring patterns. The prevalence of disc pathology was calculated. RESULTS: In all, 807 discs were injected, and 404 concordant pain responses (50%) were elicited. Three or more abnormal disc levels were identified in more than half of the patients. Complications developed in four patients (2.3%). No further complications were reported. Surgical treatment was indicated as viable in only 35 studies. CONCLUSIONS: Discography is a safe and valuable diagnostic procedure showing characteristic pain patterns that may have clinical significance. In more than half of the studies, three or more levels were identified as pain generators, suggesting that treatment decisions based on information from fewer discs injected during discography may be tenuous. PMID- 10828921 TI - Influence of occupational factors on the relation between socioeconomic status and self-reported back pain in a population-based sample of German adults with back pain. AB - STUDY DESIGN: Population-based cross-sectional postal survey and interview substudy. OBJECTIVES: To examine the association between socioeconomic status and severe back pain and to determine whether this association can be explained by occupational factors. SUMMARY OF BACKGROUND DATA: Like other disorders, back pain and its consequences are inversely related to indicators of high socioeconomic status. METHODS: The associations between indicators of socioeconomic status and presence or severity of current back pain (no back pain or back pain of low intensity and low disability versus back pain with high intensity and/or high disability) were investigated in a survey among German adults 25 to 74 years of age (n = 2731) and an interview substudy of 770 participants with a recent history of back pain.- RESULTS: In the survey, educational level was inversely associated with back pain and severe current back pain. Similarly, in the interview substudy, educational level, vocational training, occupational class, household income, and health insurance status were inversely related to severe current back pain. Age-adjusted and gender-adjusted odds ratios were 0.36 (95% confidence interval [CI] 0.25-0.52) for immediate educational level and 0.37 (95% CI 0.18-0.73) for high educational level. Recalled work tasks at the onset of back pain were significant risk factors of severe current back pain (heavy physical work: odds ratio [OR] 1.77, 95% CI 1.06-2.93; work in bent position among males: OR 1.89, 95% CI 1.03-3.46). After adjusting for occupational class or work tasks, the association between educational level and severe current back pain remained unchanged.- CONCLUSIONS: The findings support the hypothesis that severe back pain is less prevalent among adults of higher socioeconomic status. The underlying mechanism could not be explained by differences in self-reported occupational factors. PMID- 10828922 TI - Cost-benefit of muscle cocontraction in protecting against spinal instability. AB - STUDY DESIGN: Lifting dynamics and electromyographic activity were evaluated using a biomechanical model of spinal equilibrium and stability to assess cost benefit effects of antagonistic muscle cocontraction on the risk of stability failure. OBJECTIVES: To evaluate whether increased biomechanical stability associated with antagonistic cocontraction was capable of stabilizing the related increase in spinal load. SUMMARY OF BACKGROUND DATA: Antagonistic cocontraction contributes to improved spinal stability and increased spinal compression. For cocontraction to be considered beneficial, stability must increase more than spinal load. Otherwise, it may be possible for cocontraction to generate spinal loads that cannot be stabilized. METHODS: A biomechanical model was developed to compute spinal load and stability from measured electromyography and motion dynamics. As 10 healthy men performed sagittal lifting tasks, trunk motion, reaction loads, and electromyographic activities of eight trunk muscles were recorded. Spinal load and stability were evaluated as a function of cocontraction and trunk flexion angle. Stability was quantified in terms of the maximum spinal load the system could stabilize. RESULTS: Cocontraction was associated with a 12% to 18% increase in spinal compression and a 34% to 64% increase in stability. Spinal load and stability increased with trunk flexion. CONCLUSIONS: Despite increases in spinal load that had to be stabilized, the margin between stability and spinal compression increased significantly with cocontraction. Antagonistic cocontraction was found to be most beneficial at low trunk moments typically observed in upright postures. Similarly, empirically measured antagonistic cocontraction was recruited less in high-moment conditions and more in low-moment conditions. PMID- 10828924 TI - Lumbar-pelvic coordination is influenced by lifting task parameters. AB - STUDY DESIGN: Low back kinematics, including relative lumbar and pelvic motions, were quantified during controlled lifting tasks. OBJECTIVES: To evaluate the influence of load and lifting velocity on lumbar-pelvic (LP) coordination. SUMMARY OF BACKGROUND DATA: Sagittal trunk extension is achieved through the coordinated motion of the pelvis and lumbar spine. There are no data to indicate whether lifting task design influences lumbar-pelvic coordination. METHODS: Lumbar and pelvic motions were recorded from 18 healthy subjects while performing isokinetic lifting tasks of 0.1 kg and 10 kg. Coordinated motions of the pelvis (sacral spine) and low-thoracic spine were evaluated using eigenvector analyses and a ratio of lumbar and pelvic angles (L/P). RESULTS: Eigenvector models of the lumbar-pelvic coordination accurately represented empirical coordination profiles. Weight significantly influenced lumbar-pelvic coordination. Trunk extension velocity demonstrated a small but statistically significant influence on lumbar-pelvic coordination. Weight and trunk flexion angle significantly influenced lumbar/pelvic angle ratios. CONCLUSIONS: Trunk extension was achieved through simultaneous but nonlinear contributions from both the pelvis and lumbar spine throughout the range of motion. The lumbar spine accounted for 70% of the total, with increased pelvic contributions in flexed postures. Task weight increased the lumbar contribution to total trunk motion. When performing clinical evaluations of spinal kinematics, it is necessary to recognize that unloaded motions may not fully represent loaded behavior of spinal coordination. PMID- 10828923 TI - Sensitivity and specificity of the indicators of sincere effort of the EPIC lift capacity test on a previously injured population. AB - STUDY DESIGN: Randomized control post-test only. OBJECTIVES: To investigate the reliability and validity of the EPIC Lift Capacity test's indicators of sincere effort. SUMMARY OF BACKGROUND DATA: The EPIC Lift Capacity test (ELC) (Employment Potential Improvement Corp., Santa Ana, CA) is a functional evaluation tool used to identify physical limitations involved in lifting and manual materials handling. Identification of insincere effort is an integral component of such functional testing because of the potential secondary gain issues surrounding the various populations typically involved in this form of testing. The purpose of this study was to determine the sensitivity and specificity of the "indicators of sincere effort" of the EPIC Lift Capacity test when used on a previously injured population typical of subjects for which the test is designed. METHODS: Subjects consisted of 41 volunteers (age 22 to 58 years) with a previously diagnosed musculoskeletal pathology of the spine or extremities. Volunteers were randomized into either the control group, instructed to give a sincere maximum effort, or the experimental group, instructed to give an insincere effort at 50% of their perceived maximum effort. All tests were administered by certified clinical evaluators according to the standardized EPIC Lift Capacity test protocol.- RESULTS: Overall accuracy in identifying participants' level of effort was 86.84%. The indicators of valid effort exhibited both high positive (94.44%) and negative (80.00%) predictive values. The indicators of valid effort accounted for 94.9% of the total variance in the determination of the subjects' overall effort level. Interrater reliability for agreement of subjects' overall effort was good (interclass correlation coefficient = 0.82). CONCLUSIONS: Through use of standardized indicators of sincere effort, certified EPIC Lift Capacity test evaluators were able to predict sincerity of effort with a high degree of reliability and validity. The rater's systematicobservational evaluation of effort was shown to be the single best indicator of sincere effort. PMID- 10828925 TI - A controlled trial of an educational pamphlet to prevent disability after occupational low back injury. AB - STUDY DESIGN: Randomized controlled trial. OBJECTIVE: To test the ability of an educational pamphlet to improve recovery in terms of pain, work status, and health care utilization after occupational low back injury. BACKGROUND: Low back pain and disability persist as occupational health problems of epidemic proportions. Because interventions based on biomechanical models have had limited impact, recent educational approaches to preventing back problems have stressed psychosocial recovery issues. METHODS: A pamphlet was developed by compiling activity resumption, self-care, and attitudinal advice from recent publications. The pamphlet was sent at random to half of all consenting workers reporting back pain within 11 days of occupational injury between 7/96 and 6/97. Three and 6 months later, back pain, work status, health care use, and pamphlet impact outcomes were assessed through structured telephone interviews. RESULTS: Of the 726 eligible workers, 486 consented to participate. Consenters and nonconsenters and intervention and control groups were similar in initial demographic variables. The pamphlet had no statistically significant impact at the 0.05 significance level on pain severity or reduction, health care visits, or work absence. Of the 229 pamphlet recipients, 129 thought it had provided useful information, but only 25 thought it had helped them return to work more quickly. CONCLUSIONS: In this trial, a pamphlet stressing psychosocial recovery issues did not prevent or reduce postinjury pain, health care use, or work absence. PMID- 10828926 TI - Lumbar spinal stenosis: conservative or surgical management?: A prospective 10 year study. AB - STUDY DESIGN: A cohort of 100 patients with symptomatic lumbar spinal stenosis, characterized in a previous article, were given surgical or conservative treatment and followed for 10 years. OBJECTIVES: To identify the short- and long term results after surgical and conservative treatment, and to determine whether clinical or radiologic predictors for the treatment result can be defined. SUMMARY OF BACKGROUND DATA: Surgical decompression has been considered the rational treatment. However, clinical experience indicates that many patients do well with conservative treatment. METHODS: In this study, 19 patients with severe symptoms were selected for surgical treatment and 50 patients with moderate symptoms for conservative treatment, whereas 31 patients were randomized between the conservative (n = 18) and surgical (n = 13) treatment groups. Pain was decisive for the choice of treatment group. All patients were observed for 10 years by clinical evaluation and questionnaires. The results, evaluated by patient and physician, were rated as excellent, fair, unchanged, or worse. RESULTS: After a period of 3 months, relief of pain had occurred in most patients. Some had relief earlier, whereas for others it took 1 year. After a period of 4 years, excellent or fair results were found in half of the patients selected for conservative treatment, and in four fifths of the patients selected for surgery. Patients with an unsatisfactory result from conservative treatment were offered delayed surgery after 3 to 27 months (median, 3.5 months). The treatment result of delayed surgery was essentially similar to that of the initial group. The treatment result for the patients randomized for surgical treatment was considerably better than for the patients randomized for conservative treatment. Clinically significant deterioration of symptoms during the final 6 years of the follow-up period was not observed. Patients with multilevel afflictions, surgically treated or not, did not have a poorer outcome than those with single-level afflictions. Clinical or radiologic predictors for the final outcome were not found. There were no dropouts, except for 14 deaths. CONCLUSIONS: The outcome was most favorable for surgical treatment. However, an initial conservative approach seems advisable for many patients because those with an unsatisfactory result can be treated surgically later with a good outcome. PMID- 10828927 TI - Lumbar interbody fusion using the Brantigan I/F cage for posterior lumbar interbody fusion and the variable pedicle screw placement system: two-year results from a Food and Drug Administration investigational device exemption clinical trial. AB - STUDY DESIGN: A carbon fiber-reinforced polymer cage implant filled with autologous bone was designed to separate the mechanical and biologic functions of posterior lumbar interbody fusion. OBJECTIVES: To test the safety and efficacy of the carbon cage with pedicle screw fixation in a 2-year prospective study performed at six centers under a protocol approved by the Food and Drug Administration, and to present the data supporting the Food and Drug Administration approved indications. SUMMARY OF BACKGROUND DATA: The success of posterior lumbar interbody fusion has been limited by mechanical and biologic deficiencies of the donor bone. Some failures of pedicle screw fixation may be attributable to the absence of adequate load sharing through the anterior column. Combining an interbody fusion device with pedicle screw fixation may address some limitations of posterior lumbar interbody fusion or pedicle screw fixation in cases that are more complex mechanically. METHODS: This clinical study of posterior lumbar interbody fusion with pedicle screw fixation involved a prospective group of 221 patients. RESULTS: Fusion success was achieved in 176 (98.9%) of 178 patients. In the management of degenerative disc disease in patients with prior failed discectomy surgery, clinical success was achieved in 79 (86%) of 92 patients, and radiographic bony arthrodesis in 91 (100%) of 91 patients. Disc space height, averaging 7.9 mm before surgery, was increased to 12.3 mm at surgery and maintained at 11.7 mm at 2 years. Fusion success was notdiminished over multiple fusion levels. These results were significantly better than those reported in prior literature. Although significant surgical complications occurred, those attributable to the implant devices occurred less frequently and generally were minor. CONCLUSIONS: The Brantigan I/F Cage for posterior lumbar interbody fusion and the Variable Screw Placement System are safe and effective for the management of degenerative disc disease. PMID- 10828928 TI - Harvesting the intact cadaveric cervical spine (C0-Th1). AB - STUDY DESIGN: Technical report on harvesting method for human cadaveric cervical spine. OBJECTIVES: Description of a new method for harvesting the intact cervical spine during routine autopsies, including the atlanto-occipital and cervico thoracic joints, without visible disfigurement above the suprasternal notch. SUMMARY OF BACKGROUND DATA: Despite the need for cervical spine specimens, there are only few articles describing procedures for harvesting an intact cervical spine. Presently available techniques either do not preserve the atlanto occipital joint or leave visible disfigurement. METHODS: The body was placed in a prone position with the head flexed, and a posterior midline incision was performed. The spine was separated from surrounding tissue, then the caudal end was cut off through the Th1/Th2 disc space. A circular craniotomy provided access to the cranial base. A square window surrounding the foramen magnum was cut at the cranial base (through the sella turcica, the internal occipital protuberance, and 5 cm parasagittal on either side), and the entire cervical spine extracted through the posterior incision. The defect was reconstructed using wood and plaster materials. RESULTS: Eighteen specimens were harvested to date using this method. The average time of harvesting the cervical spine was less than 30 minutes. Reconstruction using wood and plaster resulted in a nearly normal appearance of the neck. CONCLUSIONS: Using this technique, the nuchal ligament providing stability to the cervical spine can be preserved. The suggested method was found simple, efficient, and reproducible for harvesting the intact cervical spine, including the atlanto-occipital and cervico-thoracic joints, from any routine autopsy. PMID- 10828929 TI - Neonatal infectious spondylitis of the cervical spine presenting with quadriplegia: a case report. AB - STUDY DESIGN: A case report.- OBJECTIVE: To highlight the evaluation and treatment of neonatal infectious spondylitis of the cervical spine. SUMMARY OF BACKGROUND DATA: Most authors advise intravenous antibiotics as first-choice treatment. The place of aspiration or operative drainage is debated, as is the position and duration of immobilization. METHODS: A 3-week-old neonate was presented with intermittent quadriplegia. RESULTS: Additional investigation demonstrated an osteolytic process in the body of C3 with a large epidural abscess compressing the spinal cord. Because an infectious spondylitis of C3 was suspected, aspiration of the abscess was performed, and antibiotic therapy was started. The patient improved to neurologically normal within 3 weeks and remains asymptomatic throughout a follow-up period of 7 years. CONCLUSIONS: Neonatal infectious spondylitis should be diagnosed early and treated promptly; otherwise, it may have devastating consequences. PMID- 10828930 TI - Rosai-Dorman disease causing cervical myelopathy. AB - STUDY DESIGN: A case report of a rare disease entity and review of the literature. OBJECTIVES: To illustrate the occurrence of an unusual disease affecting the spine and spinal canal. SUMMARY OF BACKGROUND DATA: Rosai-Dorman disease is considered an idiopathic benign lymphoproliferative disease that typically occurs in the earlier decades of life. The usual manifestation is painless massive cervical lymphadenopathy, although involvement of many extra nodal sites is common. This case illustrates Rosai-Dorman disease occurring in an elderly man with myelopathy and mass lesions of the cervical, thoracic, and lumbar spinal canal without typical lymphadenopathy.- METHODS: Case report illustrating clinical presentation as well as radiographic and pathologic findings, including comparisons to cases previously reported. RESULTS: Surgical decompression with incomplete resection of the lesion was performed, providing diagnosis and treatment guidance. The patient experienced significant neurologic improvement of myelopathy. CONCLUSIONS: Recognizing clinical and laboratory features of this disease may permit earlier diagnosis and limit or avoid surgical intervention in some cases. PMID- 10828931 TI - Femoral artery ischemia during spinal scoliosis surgery detected by posterior tibial nerve somatosensory-evoked potential monitoring. AB - STUDY DESIGN: A case report of unilateral leg ischemia caused by femoral artery compression detected using posterior tibial nerve somatosensory-evoked potentials during spinal scoliosis instrumentation surgery. OBJECTIVES: To report a rare cause of intraoperative unilateral loss of all posterior tibial nerve somatosensory-evoked potential waveforms. SUMMARY OF BACKGROUND DATA: Failure to obtain adequate popliteal fossa, spinal, subcortical, and cortical potentials during posterior tibial nerve somatosensory-evoked potential spinal cord monitoring usually results from technical factors or chronic conditions affecting the peripheral nerve. METHODS: A 16-year-old boy with thoracic scoliosis had normal posterior tibial nerve somatosensory-evoked potentials both before surgery and in the operating room immediately after anesthesia induction and prone positioning on a four-post spinal frame. RESULTS: One hour after the start of surgery, a minimal amplitude reduction of the right popliteal fossa potentials appeared. Fifteen minutes later, the amplitudes of the popliteal fossa, subcortical, and cortical potentials evoked by right posterior tibial nerve stimulation became substantially reduced. Subsequently, all waveforms were lost. Malfunction of the right posterior tibial nerve stimulator was initially suspected, but when proper function was verified, a search for other causes of this loss led to discovery of leg ischemia. The patient was repositioned on the spinal frame, and all posterior tibial nerve somatosensory-evoked potentials waveforms began to reappear 7 minutes later. There was no postoperative clinically detectable complication. CONCLUSIONS: Although technical malfunction should always be suspected when all intraoperative somatosensory-evoked potential waveforms are initially seen and subsequently lost, one should also consider the possibility that intraoperative ischemia due to limb positioning could be the etiology. PMID- 10828933 TI - Imagery PMID- 10828932 TI - Letter to the editor PMID- 10828934 TI - The effect of oral nutritional supplements on habitual dietary quality and quantity in frail elders. AB - BACKGROUND: Frail institutionalized elders have a high prevalence of nutritional risk factors, undernutrition, weight loss, and nutrition-related morbidity and excess mortality. Little information is available on effective means to intervene in this setting. HYPOTHESES: We tested the hypothesis that addition of multinutrient oral supplements to the diet of frail elders would improve their overall nutritional status and functional level. METHODS: Fifty nursing home residents aged 88+/-1 yr. were followed for 10 weeks in the course of a randomized controlled trial of supplementation with a multinutrient liquid supplement vs. a non-nutritive placebo drink. Three-day food weighing was used to analyze their habitual dietary intake before and during the final week of the intervention. Nutritional status was further assessed with nutritional biochemistries, anthropometric measurements, and body composition analysis as well as physical and functional performance tests. RESULTS: The nutritional supplement was consumed with high compliance, but did not significantly augment total caloric intake. Supplementation was associated with significant reductions in total energy, protein, fat, water, fiber, and many vitamins and minerals in the habitual diet of these nursing home residents. Nutritional status improved in terms of folate levels in serum, but no other measured vitamin or mineral indices. Body composition analysis revealed a small gain in weight, increases in fat stores, but no improvement in lean tissue mass associated with supplemention. No physical performance or functional gains were associated with supplementation. CONCLUSION: Short-term nutritional supplementation in elders at nutritional risk is offset by simultaneous reduction in voluntary food intake. It seems likely that changing other components of energy expenditure such as physical activity levels or basal metabolism may be required to produce overall improvements in nutritional intake in this setting. PMID- 10828935 TI - Vitamins and trace elements in home parenteral nutrition patients. AB - AIMS: to study the micronutrient status in home parenteral nutrition (HPN) patients and its relationship to inflammatory markers, and clinical outcome. METHODS: Vitamins (A, 25OH D3, E, B12), serum folic acid, as well as trace elements (selenium, zinc, copper, iron and manganese) were measured in 22 adult HPN patients and 14 controls. They were compared to serum malondialdehyde (MDA) concentration (as a marker of lipid peroxidation), erythrocyte superoxide dismutase (SOD) and gluthatione peroxidase (GSHPx), inflammatory markers, and clinical outcome. RESULTS: In HPN patients MDA concentration was increased whereas vitamin E concentrations were decreased, and significantly negatively correlated to MDA. Erythrocyte GSHPx and plasma selenium were decreased in the patients and positively correlated to each other. By contrast, manganese concentration was significantly increased in HPN patients and correlated to inflammatory markers. CONCLUSIONS: Adult HPN patients showed increased lipid peroxidation. This seems principally the result of low vitamin E status. In addition, these patients presented often a decrease in plasma selenium responsible for low GSHPx activity. These combined antioxidant system deficiencies contribute probably to peroxidative damage in HPN patients. Increased manganese concentrations, in view of their potential neurotoxicity have to be closely surveyed. In HPN patients micronutrient status needs regular monitoring in regard to the possibility of vitamin and/or trace element abnormalities. PMID- 10828936 TI - Effect of decreased mobility on body composition in patients with Alzheimer's disease. AB - Randomly selected 50 patients with the diagnosis of probable Alzheimer's disease, hospitalized for long-term care in a Special Care Dementia unit, were examined. None of the patients were clinically malnourished although several had low cholesterol levels. The mean lean body mass, measured by bioelectrical impedance plethysmography, was 62.5% of total body mass. The average calorie intake was 2125+398 Kcal/day, ranging from 1300 to 2900 Kcal/day, and the body weight of most subjects was stable, with the average gain of 1 lbs in the previous three months. Eighteen patients ambulated independently, 14 required assistance, and 18 were non-ambulatory. The lean body mass index was associated with the patient's age and mobility status. These results indicate that patients with advanced dementia and compromised mobility have decreased muscle mass that may result in weight loss even in the absence of malnutrition. PMID- 10828937 TI - Nutritional support in elderly patients. AB - There are now many studies which have found that undernutrition is prevalent and often unrecognized in patients admitted to hospitals and institutions. There is also evidence which links protein-energy undernutrition or its markers with clinical outcomes in acute and non-acute hospital settings and that nutritional supplements can improve outcomes in some of these settings. Active nutritional support following the catabolic phase of acute illness and extending during the rehabilitation period may be of particular benefit in improving nutritional intake, status and or outcome. A randomized controlled trial is therefore needed to test this hypothesis. PMID- 10828938 TI - Nutritional supplementation in elderly people during the course of catabolic illnesses. AB - The benefit of nutritional support applied to elderly subjects during the course of catabolic illnesses is here discussed through the results of both a nutritional intervention study and a pragmatic prospective study. The early recognition of the nutritional risk is the basis of an adequate nutrition support. We have developed a visual method for estimating meal intake during hospitalisation. It permitted the exact quantification of energy and protein intake by a dietician or immediate but visual rough estimation by the nursing staff. These latter allowed classifying the patients according to the severity of the nutritional intake impairment. The oral nutritional intervention permitted to actually increase the mean dietary intake of critically ill elderly patients. This intervention was associated to a decrease in incidence rate of grade I pressure ulcer (erythema), but did not modify the death rate. On the other hand, the continuous training of the nursing staff for nutritional risk evaluation and oral nutritional support implementation led to significant improvements in practices. Such improvements in routine care were not followed by a significant decrease in inhospital adverse outcomes. However, in both studies the resulting average intake of these patients remained far from optimal and patients specific conditions played a critical role in adverse outcome rate. Thus, we propose to support the critically ill patients using oral supplementation (which is usually safe, when they are able to eat) on the basis of nutritional risk recognition. PMID- 10828939 TI - Chemical basis for enzyme catalysis. PMID- 10828940 TI - Glutamate 189 of the D1 polypeptide modulates the magnetic and redox properties of the manganese cluster and tyrosine Y(Z) in photosystem II. AB - Recent models for water oxidation in photosystem II postulate that the tyrosine Y(Z) radical, Y(Z)(*), abstracts both an electron and a proton from the Mn cluster during one or more steps in the catalytic cycle. This coupling of proton- and electron-transfer events is postulated to provide the necessary driving force for oxidizing the Mn cluster in its higher oxidation states. The formation of Y(Z)(*) requires the deprotonation of Y(Z) by His190 of the D1 polypeptide. For Y(Z)(*) to abstract both an electron and a proton from the Mn cluster, the proton abstracted from Y(Z) must be transferred rapidly from D1-His190 to the lumenal surface via one or more proton-transfer pathways. The proton acceptor for D1 His190 has been proposed to be either Glu189 of the D1 polypeptide or a group positioned by this residue. To further define the role of D1-Glu189, 17 D1-Glu189 mutations were constructed in the cyanobacterium Synechocystis sp. PCC 6803. Several of these mutants are of particular interest because they appear to assemble Mn clusters in 70-80% of reaction centers in vivo, but evolve no O(2). The EPR and electron-transfer properties of PSII particles isolated from the D1 E189Q, D1-E189L, D1-E189D, D1-E189N, D1-E189H, D1-E189G, and D1-E189S mutants were examined. Intact PSII particles isolated from mutants that evolved no O(2) also exhibited no S(1) or S(2) state multiline EPR signals and were unable to advance beyond an altered Y(Z)(*)S(2) state, as shown by the accumulation of narrow "split" EPR signals under multiple turnover conditions. In the D1-E189G and D1-E189S mutants, the quantum yield for oxidizing the S(1) state Mn cluster was very low, corresponding to a > or =1400-fold slowing of the rate of Mn oxidation by Y(Z)(*). In Mn-depleted D1-Glu189 mutant PSII particles, charge recombination between Q(A)(*)(-) and Y(Z)(*) in the mutants was accelerated, showing that the mutations alter the redox properties of Y(Z) in addition to those of the Mn cluster. These results are consistent with D1-Glu189 participating in a network of hydrogen bonds that modulates the properties of both Y(Z) and the Mn cluster and are consistent with proposals that D1-Glu189 positions a group that accepts a proton from D1-His190. PMID- 10828941 TI - Alzheimer's disease amyloid propagation by a template-dependent dock-lock mechanism. AB - Amyloid plaques composed of the peptide Abeta are an integral part of Alzheimer's disease (AD) pathogenesis. We have modeled the process of amyloid plaque growth by monitoring the deposition of soluble Abeta onto amyloid in AD brain tissue or synthetic amyloid fibrils and show that it is mediated by two distinct kinetic processes. In the first phase, "dock", Abeta addition to the amyloid template is fully reversible (dissociation t(1/2) approximately 10 min), while in the second phase, "lock", the deposited peptide becomes irreversibly associated (dissociation t(1/2) >> 1000 min) with the template in a time-dependent manner. The most recently deposited peptide dissociates first while Abeta previously deposited becomes irreversibly "locked" onto the template. Thus, the transition from monomer to neurotoxic amyloid is mediated by interaction with the template, a mechanism that has also been proposed for the prion diseases. Interestingly, two Abeta peptides bearing primary sequence alterations implicated in heritable Abeta amyloidoses displayed faster lock-phase kinetics than wild-type Abeta. Inhibiting the initial weak docking interaction between depositing Abeta and the template is a viable therapeutic target to prevent the critical conformational transition in the conversion of Abeta((solution)) to Abeta((amyloid)) and thus prevent stable amyloid accumulation. While thermodynamics suggest that inhibiting amyloid assembly would be difficult, the present study illustrates that the protein misfolding diseases are kinetically vulnerable to intervention. PMID- 10828942 TI - Three-dimensional structures of the free and antigen-bound Fab from monoclonal antilysozyme antibody HyHEL-63(,). AB - Antigen-antibody complexes provide useful models for studying the structure and energetics of protein-protein interactions. We report the cloning, bacterial expression, and crystallization of the antigen-binding fragment (Fab) of the anti hen egg white lysozyme (HEL) antibody HyHEL-63 in both free and antigen-bound forms. The three-dimensional structure of Fab HyHEL-63 complexed with HEL was determined to 2.0 A resolution, while the structure of the unbound antibody was determined in two crystal forms, to 1.8 and 2.1 A resolution. In the complex, 19 HyHEL-63 residues from all six complementarity-determining regions (CDRs) of the antibody contact 21 HEL residues from three discontinuous polypeptide segments of the antigen. The interface also includes 11 bound water molecules, 3 of which are completely buried in the complex. Comparison of the structures of free and bound Fab HyHEL-63 reveals that several of the ordered water molecules in the free antibody-combining site are retained and that additional waters are added upon complex formation. The interface waters serve to increase shape and chemical complementarity by filling cavities between the interacting surfaces and by contributing to the hydrogen bonding network linking the antigen and antibody. Complementarity is further enhanced by small (<3 A) movements in the polypeptide backbones of certain antibody CDR loops, by rearrangements of side chains in the interface, and by a slight shift in the relative orientation of the V(L) and V(H) domains. The combining site residues of complexed Fab HyHEL-63 exhibit reduced temperature factors compared with those of the free Fab, suggesting a loss in conformational entropy upon binding. To probe the relative contribution of individual antigen residues to complex stabilization, single alanine substitutions were introduced in the epitope of HEL recognized by HyHEL-63, and their effects on antibody affinity were measured using surface plasmon resonance. In agreement with the crystal structure, HEL residues at the center of the interface that are buried in the complex contribute most to the binding energetics (DeltaG(mutant) - DeltaG(wild type) > 3.0 kcal/mol), whereas the apparent contributions of solvent-accessible residues at the periphery are much less pronounced (<1.5 kcal/mol). In the latter case, the mutations may be partially compensated by local rearrangements in solvent structure that help preserve shape complementarity and the interface hydrogen bonding network. PMID- 10828943 TI - Three-dimensional structure of rat surfactant protein A trimers in association with phospholipid monolayers. AB - Surfactant protein A (SP-A) is a C-type lectin found primarily in the lung and plays a role in innate immunity and the maintenance of surfactant integrity. To determine the three-dimensional (3D) structure of SP-A in association with a lipid ligand, we have used single particle electron crystallography and computational 3D reconstruction in combination with molecular modeling. Recombinant rat SP-A, containing a deletion of the collagen-like domain, was incubated with dipalmitoylphosphatidylcholine:egg phosphatidylcholine (1:1, wt/wt) lipid monolayers in the presence of calcium, negatively stained, and examined by transmission electron microscopy. Images of SP-A-lipid complexes with different angular orientations were used to reconstruct the 3D structure of the protein. These results showed that SP-A subunits readily formed trimers and interacted with lipid monolayers exclusively via the globular domains. A homology based molecular model of SP-A was generated and fitted into the electron density map of the protein. The plane of the putative lipid-protein interface was relatively flat and perpendicular to the hydrophobic neck region, and the cleft region in the middle of the trimer had no apparent charge clusters. Amino acid residues that are known to affect lipid interactions, Glu(195) and Arg(197), were located at the protein-lipid interface. The molecular model indicated that the hydrophobic neck region of the SP-A did not interact with lipid monolayers but was instead involved in intratrimeric subunit interactions. The glycosylation site of SP-A was located at the side of each subunit, suggesting that the covalently linked carbohydrate moiety probably occupies the spaces between the adjacent globular domains, a location that would not sterically interfere with ligand binding. PMID- 10828944 TI - Location of cyanine-3 on double-stranded DNA: importance for fluorescence resonance energy transfer studies. AB - Fluorescence resonance energy transfer provides valuable long-range distance information about macromolecules in solution. Fluorescein and Cy3 are an important donor-acceptor pair of fluorophores; the characteristic Forster length for this pair on DNA is 56 A, so the pair can be used to study relatively long distances. Measurement of FRET efficiency for a series of DNA duplexes terminally labeled with fluorescein and Cy3 suggests that the Cy3 is close to the helical axis of the DNA. An NMR analysis of a self-complementary DNA duplex 5'-labeled with Cy3 shows that the fluorophore is stacked onto the end of the helix, in a manner similar to that of an additional base pair. This provides a known point from which distances calculated from FRET measurements are measured. Using the FRET efficiencies for the series of DNA duplexes as restraints, we have determined an effective position for the fluorescein, which is maximally extended laterally from the helix. The knowledge of the fluorophore positions can now be used for more precise interpretation of FRET data from nucleic acids. PMID- 10828945 TI - Structures of three inhibitor complexes provide insight into the reaction mechanism of the human methylenetetrahydrofolate dehydrogenase/cyclohydrolase. AB - Enzymes involved in tetrahydrofolate metabolism are of particular pharmaceutical interest, as their function is crucial for amino acid and DNA biosynthesis. The crystal structure of the human cytosolic methylenetetrahydrofolate dehydrogenase/cyclohydrolase (DC301) domain of a trifunctional enzyme has been determined previously with a bound NADP cofactor. While the substrate binding site was identified to be localized in a deep and rather hydrophobic cleft at the interface between two protein domains, the unambiguous assignment of catalytic residues was not possible. We succeeded in determining the crystal structures of three ternary DC301/NADP/inhibitor complexes. Investigation of these structures followed by site-directed mutagenesis studies allowed identification of the amino acids involved in catalysis by both enzyme activities. The inhibitors bind close to Lys56 and Tyr52, residues of a strictly conserved motif for active sites in dehydrogenases. While Lys56 is in a good position for chemical interaction with the substrate analogue, Tyr52 was found stacking against the inhibitors' aromatic rings and hence seems to be more important for proper positioning of the ligand than for catalysis. Also, Ser49 and/or Cys147 were found to possibly act as an activator for water in the cyclohydrolase step. These and the other residues (Gln100 and Asp125), with which contacts are made, are strictly conserved in THF dehydrogenases. On the basis of structural and mutagenesis data, we propose a reaction mechanism for both activities, the dehydrogenase and the cyclohydrolase. PMID- 10828946 TI - The caa(3) terminal oxidase of Rhodothermus marinus lacking the key glutamate of the D-channel is a proton pump. AB - The thermohalophilic bacterium Rhodothermus marinus expresses a caa(3)-type dioxygen reductase as one of its terminal oxidases. The subunit I amino acid sequence shows the presence of all the essential residues of the D- and K-proton channels, defined in most heme-copper oxidases, with the exception of the key glutamate residue located in the middle of the membrane dielectric (E278 in Paracoccus denitrificans). On the basis of homology modeling studies, a tyrosine residue (Y256, R. marinus numbering) has been proposed to act as a functional substitute [Pereira, M. M., Santana, M., Soares, C. M., Mendes, J., Carita, J. N., Fernandes, A. S., Saraste, M., Carrondo, M. A., and Teixeira, M. (1999) Biochim. Biophys. Acta 1413, 1-13]. Here, R. marinus caa(3) oxidase was reconstituted in liposomes and shown to operate as a proton pump, translocating protons from the cytoplasmic side of the bacterial inner membrane to the periplasmatic space with a stoichiometry of 1H(+)/e(-), as in the case in heme copper oxidases that contain the glutamate residue. Possible mechanisms of proton transfer in the D-channel with the participation of the tyrosine residue are discussed. The observation that the tyrosine residue is conserved in several other members of the heme-copper oxidase superfamily suggests a common alternative mode of action for the D-channel. PMID- 10828947 TI - Fluorescence decay and spectral evolution in intact photosystem I of higher plants. AB - A photosystem I preparation from maize, containing its full antenna complement (PSI-200) and in which detergent effects on chlorophyll coupling are almost completely absent, has been studied by time-resolved fluorescence techniques with approximately 5 ps resolution at 280 and 170 K in the wavelength interval of 690 780 nm. The data have been analyzed in terms of both the decay-associated spectra (DAS) and the time-resolved emission spectra (TRES). As in a previous room temperature study [Turconi, S., Weber, N., Schweitzer, D., Strotmann, H., and Holzwarth, A. R. (1994) Biochim. Biophys. Acta 1187, 324-334], the 280 K decay is well described by three DAS components in the 11-130 ps time range, the fastest of which displays both positive and negative amplitudes characteristic of excitation transfer from the bulk to the red antenna forms. Both the 57 and 130 ps components have all positive amplitudes and describe complex decay and equilibration processes involving the red forms. At 170 K, four major components in the 10-715 ps time range are required to describe the decay. The fastest represents bulk to red form transfer processes, while the 55, 216, and 715 ps decays, with all positive amplitudes, have maxima near 720, 730, and 740 nm, respectively, in accord with previous steady-state fluorescence measurements. The width and asymmetry of these DAS indicate that they are spectrally complex and represent decay and equilibration processes involving the red forms. Spectral evolution during the fluorescence decay process was analyzed in terms of the TRES. The red shifting of the TRES was analyzed in terms of the first central spectral moment (mean spectral energy) which is biexponential at both temperatures. The slower component, which describes equilibration between the red forms, leads to spectral red shifting during the entire fluorescence decay process, and the mean lifetimes of the spectral moments at 280 and 170 K (86 and 291 ps, respectively) are similar to the mean lifetimes of the fluorescence decays (119 and 384 ps, respectively). Thus, both spectral evolution and the trapping-associated fluorescence decay occur on a similar time scale, and both processes display a very similar temperature sensitivity. On the basis of these data, it is concluded that trapping in PSI-200 is to a large extent rate-limited by excitation diffusion in the antenna and in particular by the slow "uphill" transfer from the low-energy forms to the bulk and/or inner core chlorophyll molecules. PMID- 10828948 TI - Chromophore-apoprotein interactions in Synechocystis sp. PCC6803 phytochrome Cph1. AB - The secondary, tertiary, and quaternary structures of the Synechocystis Cph1 phytochrome were investigated by absorption and circular dichroism spectroscopy, size exclusion chromatography, and limited proteolysis. The Cph1 protein was coexpressed with a bacterial thioredoxin in Escherichia coli, reconstituted in vitro with tetrapyrrole chromophores, and purified by chitin affinity chromatography. The resultant Cph1 holoproteins were essentially pure and had the specific absorbance ratio (SAR) of 0.8-0.9. Circular dichroism spectroscopy and limited proteolysis showed that the chromophore binding induced marked conformational changes in the Cph1 protein. The alpha-helical content increased to 42-44% in the holoproteins from 37% in the apoprotein. However, no significant difference in the secondary structure was detected between the Pr and Pfr forms. The tertiary structure of the Cph1 apoprotein appeared to be relatively flexible but became more compact and resistant to tryptic digestion upon chromophore binding. Interestingly, a small chromopeptide of about 30 kDa was still predominant even after longer tryptic digestion. The N-terminal location of this chromopeptide was confirmed by expression in E. coli and in vitro reconstitution with chromophores of the 32.5 kDa N-terminal fragment of the Cph1 protein. This chromopeptide was fully photoreversible with the spectral characteristic similar to that of the full-size Cph1 protein. The Cph1 protein forms dimers through the C-terminal region. These results suggest that the prokaryotic Cph1 phytochrome shares the structural and conformational characteristics of plant phytochromes, such as the two-domain structure consisting of the relatively compact N-terminal and the relatively flexible C-terminal regions, in addition to the chromophore induced conformational changes. PMID- 10828949 TI - Spectroscopic and electrochemical studies on structural change of plastocyanin and its tyrosine 83 mutants induced by interaction with lysine peptides. AB - Interactions of wild-type and Tyr83 mutant (Y83F, Y83S, Y83L, and Y83H) plastocyanins (PCs) with lysine peptides as models for the PC interacting site of cytochrome f have been studied by absorption, resonance Raman, and electron paramagnetic resonance (EPR) spectroscopies and electrochemical measurements. The spectral and electrochemical properties of PCs corresponded well with each other; species having a longer wavelength maximum for the S(Cys) pi --> Cu 3d(x)()()2( )(y)()()2 charge transfer (CT) band observed around 600 nm and a stronger intensity for the 460-nm absorption band exhibited stronger intensities for the positive Met --> Cu 3d(x)()()2(-)(y)()()2 and negative His pi(1) --> Cu 3d(x)()()2(-)(y)()()2 circular dichroism (CD) bands at about 420 and 470 nm, respectively, a lower average nu(Cu)(-)(S) frequency, a smaller |A( parallel)| EPR parameter, and a higher redox potential, properties all related to a weaker Cu-S(Cys) bond and a more tetrahedral planar geometry for the Cu site. Similarly, on oligolysine binding to wild-type and several Tyr83 mutant PCs, a longer absorption maximum for the 600-nm CT band, a stronger intensity for the 460-nm absorption band, stronger 420-nm positive and 470-nm negative CD bands, and a lower average nu(Cu)(-)(S) frequency were observed, suggesting that PC assumes a slight more tetrahedral geometry on binding of oligolysine. Since changes were observed for both wild-type and Tyr83 mutant PCs, the structural change due to binding of oligolysine to PC may not be transmitted through the path of Tyr83 Cys84-copper by a cation-pi interaction which is proposed for electron transfer. PMID- 10828950 TI - Binding of O(2) and its reduction are both retarded by replacement of valine 279 by isoleucine in cytochrome c oxidase from Paracoccus denitrificans. AB - The crystal structure of the heme-copper oxidases suggested a putative channel of oxygen entry into the heme-copper site of O(2) reduction. Changing a conserved valine near this center in cytochrome bo(3) of Escherichia coli to isoleucine caused a significant increase in the apparent K(M) for oxygen with little or no change in V(max), suggesting that oxygen diffusion had been partially blocked [Riistama, S., Puustinen, A., Garcia-Horsman, A., Iwata, S., Michel, H., and Wikstrom, M. (1996) Biochim. Biophys. Acta 1275, 1-4]. To study this phenotype further using rapid kinetic methods, the corresponding change (V279I) has been made in cytochrome aa(3) from Paracoccus denitrificans. In this mutant, the apparent K(M) for oxygen is 8 times higher than in the wild-type enzyme, whereas V(max) is decreased only to approximately half of the wild-type value. Flow-flash kinetic measurements show that the initial binding of oxygen to the heme of the binuclear site is indeed much slower in the mutant than in the wild-type enzyme. However, the subsequent phases of the reaction with O(2) are also slow although the pure heme-to-heme electron transfer process is essentially unperturbed. It is suggested that the mutation sterically hinders O(2) entry into the binuclear site and that it may also perturb the structure of local water molecules involved in proton transfer to this site. PMID- 10828951 TI - Coupling of electron transfer with proton transfer at heme a and Cu(A) (redox Bohr effects) in cytochrome c oxidase. Studies with the carbon monoxide inhibited enzyme. AB - A study is presented on the coupling of electron transfer with proton transfer at heme a and Cu(A) (redox Bohr effects) in carbon monoxide inhibited cytochrome c oxidase isolated from bovine heart mitochondria. Detailed analysis of the coupling number for H(+) release per heme a, Cu(A) oxidized (H(+)/heme a, Cu(A) ratio) was based on direct measurement of the balance between the oxidizing equivalents added as ferricyanide to the CO-inhibited fully reduced COX, the equivalents of heme a, Cu(A), and added cytochrome c oxidized and the H(+) released upon oxidation and all taken up back by the oxidase upon rereduction of the metal centers. One of two reductants was used, either succinate plus a trace of mitochondrial membranes (providing a source of succinate-c reductase) or hexaammineruthenium(II) as the chloride salt. The experimental H(+)/heme a, Cu(A) ratios varied between 0.65 and 0.90 in the pH range 6.0-8.5. The pH dependence of the H(+)/heme a, Cu(A) ratios could be best-fitted by a function involving two redox-linked acid-base groups with pK(o)-pK(r) of 5.4-6.9 and 7.3-9.0, respectively. Redox titrations in the same samples of the CO-inhibited oxidase showed that Cu(A) and heme a exhibited superimposed E'(m) values, which decreased, for both metals, by around 20 mV/pH unit increase in the range 6.0 8.5. A model in which oxido-reduction of heme a and Cu(A) are both linked to the pK shifts of the two acid-base groups, characterized by the analysis of the pH dependence of the H(+)/heme a, Cu(A) ratios, provided a satisfactory fit for the pH dependence of the E'(m) of heme a and Cu(A). The results presented are consistent with a primary involvement of the redox Bohr effects shared by heme a and Cu(A) in the proton-pumping activity of cytochrome c oxidase. PMID- 10828952 TI - GCN4 binds with high affinity to DNA sequences containing a single consensus half site. AB - bZip proteins contain a bipartite DNA-binding motif consisting of a "leucine zipper" dimerization domain and a highly charged "basic region" that directly contacts DNA. These transcription factors form dimeric complexes with each monomer recognizing half of a symmetric or nearly symmetric DNA site. We have found that the bZip protein GCN4 can also bind with high affinity to DNA sites containing only a single GCN4 consensus half-site. Because several recent lines of evidence have suggested a role for monomeric DNA binding by bZip proteins, we investigated the structure of the GCN4.half-site complex. Quantitative DNA binding and affinity cleaving studies support a model in which GCN4 binds as a dimer, with one monomer making specific contacts to the consensus half-site and the other monomer forming nonspecific contacts that are nonetheless important for binding affinity. We also examined the folding transition induced in the basic regions of this complex upon binding DNA. Circular dichroism (CD) studies demonstrate that the basic regions of both monomers are helical, suggesting that a protein folding transition may be required for both specific and nonspecific DNA binding by GCN4. PMID- 10828953 TI - Chemical structure of nuclear proteins which are phosphorylated during meiotic maturation of starfish oocytes. AB - Oocytes of the starfish, Asterina pectinifera, are arrested at the G2 phase of meiosis I and possess a prominent germinal vesicle in which maternal stores of nuclear proteins which are destined for use primarily by the early embryo are stored. Germinal vesicle breakdown and subsequent oocyte maturation is triggered by activation of the p34(cdc2)/cyclin B complex, which is present as the preform in the cytoplasm. The aim of the present study was to identify and biochemically characterize in vivo substrates of the kinase. Two nucleic acid binding nuclear proteins designated NAAP1 and NAAP2 were found, both of which contain 345 amino acid residues with pI 3. 6 and which serve as substrates. The only difference between the two proteins was in the primary amino acid sequence at position 51, which is Asn in NAAP1 but Thr in NAAP2. NAAPs are phosphorylated in vivo during oocyte maturation but not at the meiotic G(2) stage. NAAPs are phosphorylated in vitro by the cdc2 kinase on the same site as in vivo. Although there are other evolutionarily conserved consensus sequences for phosphorylation by mitotically active cdc2 kinase in NAAPs and NAAP-derived fragments containing the sequences were efficiently phosphorylated in vitro, these sites in the intact NAAPs were not phosphorylated either in vivo or in vitro. These results suggest that the tertiary structure of NAAPs affects the target specificity of the cdc2 kinase. PMID- 10828954 TI - DNA binding in the central channel of bacteriophage T7 helicase-primase is a multistep process. Nucleotide hydrolysis is not required. AB - Many helicases assemble into ring-shaped hexamers and bind DNA in their central channel. This raises the question as to how the DNA gets into the central channel to form a topologically linked complex. We have used the presteady-state stopped flow kinetic method and protein fluorescence changes to investigate the mechanism of single-stranded DNA (ssDNA) binding to the bacteriophage T7 helicase-primase, gp4A'. We have found that the kinetics of 30-mer ssDNA binding to a preformed gp4A' hexamer in the presence of both Mg-dTMP-PCP and Mg-dTTP are similar, indicating that Mg-dTTP binding is sufficient and hydrolysis is not necessary for efficient DNA binding. Multiple transient changes in gp4A' fluorescence revealed a four-step mechanism for DNA binding with Mg-dTTP. These transient changes were analyzed by global fitting and kinetic simulation to determine the intrinsic rate constants of this four-step mechanism. The initial steps, including the bimolecular encounter of the DNA with the helicase and a subsequent conformational change, were fast. We propose that these initial steps of DNA binding occur at a readily accessible site, which is likely to be on the outside of the hexamer ring. The binding of the 30-mer ssDNA at this loading site is followed by slower conformational changes that allow the DNA to transit into the central channel of gp4A' via a ring-opening or threading pathway. PMID- 10828955 TI - Investigation of the proposed interdomain ribose zipper in hairpin ribozyme cleavage using 2'-modified nucleosides. AB - The hairpin ribozyme achieves catalytic cleavage through interaction of essential nucleotides located in two distinct helical domains that include internal loops. Initial docking of the two domains is ion dependent and appears to be followed by a structural rearrangement that allows the ribozyme to achieve a catalytically active state that can undergo cleavage. The proposed structural rearrangement may also be ion dependent and is now of increased importance due to recent evidence that docking is not rate limiting and that metal ions are unlikely to be involved in the chemical cleavage step. An initial structural model of the docked hairpin ribozyme included a proposal for a ribose zipper motif that involves two pairs of hydroxyl groups at A(10) and G(11) in domain A pairing with C(25) and A(24) in domain B, respectively. We have used a chemical functional group substitution technique to study whether this proposed ribose zipper is likely to be present in the active, conformationally rearranged ribozyme that is fit for cleavage. We have chemically synthesized a series of individually modified hairpin ribozymes containing 2'-analogues of nucleosides, that include 2'-deoxy and 2'-deoxy-2' fluoro at each of the four nucleoside positions, 2'-amino-2'-deoxy, 2'-deoxy-2' thio, and 2'-arabino at position C(25), and 2'-oxyamino at position A(10), as well as some double substitutions, and we studied their cleavage rates under both single- and multiple-turnover conditions. We conclude that at least some of the hydrogen-bonding interactions in the ribose zipper motif, either as originally proposed or in a recently suggested structural variation, are unlikely to be present in the active rearranged form of the ribozyme that undergoes cleavage. Instead, we provide strong evidence for a very precise conformational positioning for the residue C(25) in the active hairpin. A precise conformational requirement would be expected for C(25) if it rearranges to form a base-triple with A(9) and the essential residue neighboring the cleavage site G(+1), as recently proposed by another laboratory. Our results provide further support for conformational rearrangement as an important step in hairpin ribozyme cleavage. PMID- 10828956 TI - DNA strand transfer catalyzed by vaccinia topoisomerase: peroxidolysis and hydroxylaminolysis of the covalent protein-DNA intermediate. AB - Vaccinia topoisomerase forms a covalent DNA-(3'-phosphotyrosyl)-enzyme intermediate at sites containing the sequence 5'-CCCTT downward arrow. The covalently bound topoisomerase can religate the CCCTT strand to a 5'-OH terminated polynucleotide or else transfer the strand to a non-DNA nucleophile such a water or glycerol. Here, we report that vaccinia topoisomerase also catalyzes strand transfer to hydrogen peroxide. The observed alkaline pH dependence of peroxidolysis is consistent with enzyme-mediated attack by peroxide anion on the covalent intermediate. The reaction displays apparent first-order kinetics. From a double-reciprocal plot of k(obs) versus [H(2)O(2)] at pH 10, we determined a rate constant for peroxidolysis of 6.3 x 10(-)(3) s(-)(1). This rate is slower by a factor of 200 than the rate of topoisomerase-catalyzed strand transfer to a perfectly aligned 5'-OH DNA strand but is comparable to the rate of DNA strand transfer across a 1-nucleotide gap. Strand transfer to 2% hydrogen peroxide is 300 times faster than strand transfer to 20% glycerol and approximately 2000 times faster than topoisomerase-catalyzed hydrolysis of the covalent intermediate. Hydroxylamine is also an effective nucleophile in topoisomerase-mediated strand transfer (k(obs) = 6.4 x 10(-)(4) s(-)(1)). The rates of the peroxidolysis, hydroxylaminolysis, glycerololysis, and hydrolysis reactions catalyzed by the mutant enzyme H265A were reduced by factors of 100 700, in accordance with the 100- to 400-fold rate decrements in DNA cleavage and religation by H265A. We surmise that vaccinia topoisomerase catalyzes strand transfer to DNA and non-DNA nucleophiles via a common reaction pathway in which His-265 stabilizes the scissile phosphate in the transition state rather than acting as a general acid or base. PMID- 10828957 TI - RPA stabilizes the XPA-damaged DNA complex through protein-protein interaction. AB - The xeroderma pigmentosum group A complementing protein (XPA) and eukaryotic replication protein A (RPA) are among the major damage-recognition proteins involved in the early stage of nucleotide excision repair (NER). XPA and RPA are able to bind damaged DNA independently, although RPA interaction stimulates XPA binding to damaged DNA [Li, L., Lu, X., Peterson, C. A., and Legerski, R. J. (1995) Mol. Cell. Biol. 15, 5396-5402 (1); Stigger, E., Drissi, R., and Lee, S. H. (1998) J. Biol. Chem. 273, 9337-9343 (2)]. In this study, we used surface plasmon resonance (SPR) analysis to investigate the interaction of XPA and RPA with two major types of UV-damaged DNA: the (6-4) photoproduct and the cis-syn cyclobutane dimer of thymidine. Both XPA and RPA preferentially bind to (6-4) photoproduct-containing duplex DNA over cis-syn cyclobutane dimer-containing DNA. The binding of XPA to (6-4) photoproduct was weak (K(D) = 2.13 x 10(-)(8) M), whereas RPA showed a very stable interaction with (6-4) photoproduct (K(D) = 2. 02 x 10(-)(10) M). When XPA and RPA were incubated together, the stability of the XPA-damaged DNA interaction was significantly enhanced by wild-type RPA. On the other hand, mutant RPA (RPA:p34Delta33C) defective in its interaction with XPA failed to stabilize XPA-damaged DNA complex. Taken together, our results suggest that a role for RPA in UV-damage recognition is to stabilize XPA-damaged DNA complex through protein-protein interaction. PMID- 10828958 TI - Kinetic and structural characterization of a two-domain streptokinase: dissection of domain functionality. AB - The mammalian protease plasminogen can be activated by bacterial activators, the three-domain (alpha, beta, gamma) streptokinases and the one-domain (alpha) staphylokinases. These activators act as plasmin(ogen) cofactors, and the resulting complexes initiate proteolytic activity of host plasminogen which facilitates bacterial colonization of the host organism. We have investigated the kinetic mechanism of the plasminogen activation mediated by a novel two-domain (alpha, beta) streptokinase isolated from Streptococcus uberis (Sk(U)) with specificity toward bovine plasminogen. The interaction between Sk(U) and plasminogen occurred in two steps: (1) rapid association of the proteins and (2) slow transition to the active complex Sk(U)-PgA. The complex Sk(U)-PgA converted plasminogen to plasmin with the following parameters: K(m) < or = 1.5 microM and k(cat) = 0.55 s(-)(1). The ability of proteolytic fragments of Sk(U) to activate plasminogen was investigated. Only two C-terminal segments (97-261 and 123-261), which both contain the beta-domain (126-261), were shown to be active. They initiated plasminogen activation in complex with plasmin, but not with plasminogen, and thereby exhibited functional similarity to the staphylokinase. The fusion protein His(6)-Sk(U) (i.e., Sk(U) with a small N-terminal tag) acted exclusively in complex with plasmin as well. These observations demonstrate that (1) the N-terminal alpha-domain, including a native N-terminus, was necessary for "virgin" activation of the associated plasminogen in the Sk(U)-PgA complex and (2) the C-terminal beta-domain of Sk(U) is important for recognition of the substrate in the Sk(U)-PgA complex. PMID- 10828959 TI - Conformational difference between PDE4 apoenzyme and holoenzyme. AB - The type 4 cAMP-specific phosphodiesterases (PDE4s) are Mg(2+)-dependent hydrolases that catalyze the hydrolysis of 3', 5'-cAMP to AMP. Previous studies indicate that PDE4 exists in two conformations that bind the inhibitor rolipram with affinities differing by more than 100-fold. Here we report that these two conformations are the consequence of PDE4 binding to its metal cofactor such as Mg(2+). Using a fluorescence resonance energy transfer (FRET)-based equilibrium binding assay, we identified that L-791,760, a fluorescent inhibitor, binds to the apoenzyme (free enzyme) and the holoenzyme (enzyme bound to Mg(2+)) with comparable affinities (K(d) approximately 30 nM). By measuring the displacement of the bound L-791,760, we have also identified that other inhibitors bind differentially with the apoenzyme and the holoenzyme depending upon their structure. CDP-840, SB-207499, and RP-73401 bind preferentially to the holoenzyme. The conformational-sensitive inhibitor (R)-rolipram binds to the holoenzyme and apoenzyme with affinities (K(d)) of 5 and 300 nM, respectively. In contrast to its high affinity (K(d) approximately 2 microM) and active holoenzyme complex, cAMP binds to the apoenzyme nonproductively with a reduced affinity (K(d) approximately 170 microM). These results demonstrate that cofactor binding to PDE4 is responsible for eliciting its high-affinity interaction with cAMP and the activation of catalysis. PMID- 10828960 TI - Monoclonal antibody light chain with prothrombinase activity. AB - Prothrombin is the precursor of thrombin, a central enzyme in coagulation. Autoantibodies to prothrombin are associated with thromboembolism, but the mechanisms by which the antibodies modulate the coagulation processes are not understood. We screened a panel of 34 monoclonal antibody light chains isolated from patients with multiple myeloma for prothrombinase activity by an electrophoresis method. Two light chains with the activity were identified, and one of the light chains was characterized further. The prothrombinase activity eluted from a gel-filtration column run in denaturing solvent (6 M guanidine hydrochloride) at the characteristic positions of the light chain dimer and monomer. A constant level of catalytic activity was observed across the width of the light chain monomer peak, assessed as the cleavage of IEGR methylcoumarinamide, a peptide substrate corresponding to residues 268-271 of prothrombin. Hydrolysis of this peptide by the light chain was saturable and consistent with Michaelis-Menten-Henri kinetics (K(m) 103 microM; k(cat) of 2.62 x 10(-)(2)/min). Four cleavage sites in prothrombin were identified by N-terminal sequencing of the fragments: Arg(155)-Ser(156), Arg(271)-Thr(272), Arg(284) Thr(285), and Arg(393)-Ser(394). The light chain did not cleave radiolabeled albumin, thyroglobulin, and annexin V under conditions that readily permitted detectable prothrombin cleavage. Two prothrombin fragments (M(r) 55 000 and 38 000), were isolated by anion-exchange chromatography and were observed to cleave a thrombin substrate, tosyl-GPR-nitroanilide. Conversion of fibrinogen to fibrin was accelerated by the prothrombin fragments generated by the light chain. These finding suggest a novel mechanism whereby antibodies can induce a procoagulant state, i.e., prothrombin activation via cleavage of the molecule. PMID- 10828961 TI - Spectroscopic analysis of pH-induced changes in the molecular features of type A botulinum neurotoxin light chain. AB - Clostridial botulinum neurotoxins (BoNTs) cause neuroparalysis by blocking neurotransmitter release at the neuromuscular junctions. While the toxin's heavy chain (HC) is involved in binding and internalization, the light chain (LC) acts as a unique Zn(2+)-endopeptidase against a target protein in the exocytotic docking/fusion machinery. During the translocation of the LC to the cytosol, it is exposed to the endosomal low pH. Low pH showed a dramatic change in the BoNT/A LC polypeptide folding as indicated by differential heat denaturation. Furthermore, binding of 1-anilinonaphthalenesulfonate (ANS) revealed exposure of hydrophobic domains of BoNT/A LC at low pH. Low-pH-induced structural (and by implication the endopeptidase activity) changes were completely reversible. Exposure of BoNT/A LC to low pH (4.7) did not, however, evoke the loss of Zn(2+) bound to its active site. Implications of these observations to the delivery of active BoNT/A LC to the nerve cell are discussed. We further analyzed the nature of low-pH-induced change in the polypeptide folding of BoNT/A LC by Trp fluorescence measurements. The Trp fluorescence peak was observed at 322 nm, and the two fluorescence lifetime components estimated at 2.1 ns (88%) and 0.6 ns (12%) did not change much at low pH. These observations suggested that the two Trp residues are buried and constrained in a hydrophobic environment, and it is likely that the core of the BoNT/A LC protein matrix does not participate in the low-pH-induced structural alteration. This conclusion was further supported by the near-UV circular dichroism spectra under two pH conditions. PMID- 10828962 TI - Partially unfolded equilibrium state of hen lysozyme studied by circular dichroism spectroscopy. AB - Equilibrium unfolding of hen egg white lysozyme as a function of GdnCl concentration at pH 0.9 was studied over a temperature range 268.2-303.2 K by means of CD spectroscopy. As monitored by far- and near-UV CD at 222 and 289 nm, the lack of coincidence between two unfolding transition curves was observed, which suggests the existence of a third conformational species in addition to native and unfolded states. The three-state model, in which a stable intermediate is populated, was employed to estimate the thermodynamic parameters for the GdnCl induced unfolding. It was found that the transition from the native to intermediate states proceeds with significant changes in enthalpy and entropy due to an extremely cooperative process, while the transition from the intermediate to unfolded states shows a low cooperativity with small enthalpy and entropy changes. These results indicate that the highest energy barrier for the GdnCl induced unfolding of hen lysozyme is located in the process from the native state to the intermediate state, and this process is largely responsible for the cooperativity of protein unfolding. PMID- 10828963 TI - Inner nuclear membrane protein LBR preferentially interacts with DNA secondary structures and nucleosomal linker. AB - The lamin B receptor (LBR) is an integral protein of inner nuclear membrane whose nucleoplasmic amino-terminal domain contributes to the attachment of the membrane to chromatin. Here we analyzed the interactions of a recombinant GST protein containing the amino-terminal domain of the protein with in vitro reconstituted nucleosomes and short DNA fragments. Data show that the LBR amino-terminal domain (AT) binds linker DNA but does not interact with the nucleosome core. Titration and competition studies revealed that the interaction between LBR AT and DNA is saturable, of high affinity (K(D) approximately 4 nM), independent of DNA sequence, and enhanced by DNA curvature and supercoiling. In this respect, LBR amino-terminal domain binding to nucleosomes is similar to that of histone H1 and non histone proteins HMG1/2 which both bind preferentially to linker DNA and present a significant affinity for DNA secondary structures. PMID- 10828964 TI - Association of cytochromes P450 with their reductases: opposite sign of the electrostatic interactions in P450BM-3 as compared with the microsomal 2B4 system. AB - The role of electrostatic interactions in the association of P450s with their nicotinamide adenine dinucleotide phosphate- (NADPH) dependent flavoprotein reductases was studied by fluorescence resonance energy transfer. The fluorescent probe 7-(ethylamino)-3-(4'-maleimidylphenyl)-4-methylcoumarin maleimide (coumarylphenylmaleimide, CPM) was introduced into the flavoprotein molecule at a 1:1 molar ratio. The interaction of P450 2B4 and NADPH-P450 reductase (CPR) from rabbit liver microsomes was compared with that of the isolated heme domain (BMP) and the flavoprotein domain (BMR) of P450BM-3. The cross-pairs of the components were also studied. Increasing ionic strength (0.05-0.5 M) was shown to result in the dissociation of the CPR-P450 2B4 complex with the dissociation constant increasing from 0.01 to 0.09 microM. This behavior is consistent with the assumption that charge pairing between CPR and P450 2B4 is involved in their association. In contrast, the electrostatic component of the interaction of the partners in P450BM-3 was shown to have an opposite sign. The isolated BMP and BMR domains have very low affinity for each other and the dissociation constant of their complex decreases from 8 to 3 microM with increasing ionic strength (0.05 0.5 M). Importantly, the BMP-CPR and P450 2B4-BMR "mixed", heterogeneous pairs behave similarly to the pairs of BMP and P450 2B4 with their native electron donors. Therefore, the observed difference in the interaction mechanisms between these two systems is determined mainly by the different structure of the heme proteins rather than their flavoprotein counterparts. P450BM-3 is extremely efficient and highly coupled, with the reductase and the P450 domains tethered to one another. Therefore, in contrast to P450 2B4-CPR binding, very tight binding between the P450BM-3 redox partners would be of no value in the synchronization of complex formation during catalytic turnover. PMID- 10828965 TI - Inactivation of cysteine proteases by (acyloxy)methyl ketones using S'-P' interactions. AB - We have synthesized (acyloxy)methyl ketone inactivators of papain, cathepsin B, and interleukin-1beta conversion enzyme (ICE) that interact with both the S and S' subsites. The value of k(inact)/K(i) for these inactivators is strongly dependent on the leaving group. For example, Z-Phe-Gly-CH(2)-X is a poor inactivator of papain when X is OCOCH(3) (k(inact)/K(i) = 2.5 M(-)(1) s(-)(1)) but becomes a potent inactivator when X is OCO-L-Leu-Z (k(inact)/K(i) = 11 000 M( )(1) s(-)(1)). Since these leaving groups have similar chemical reactivities, the difference in potency must be attributed to interactions with the S' sites. The potency of the leaving group correlates with the P' specificity of papain. Similar results are also observed for the inactivation of cathepsin B by these compounds. A series of inactivators with the general structure Fmoc-L-Asp-CH(2)-X were designed to inactivate ICE. No inhibition was observed when X was OCOCH(3). In contrast, ICE is inactivated when X is OCO-D-Pro-Z (k(inact)/K(i) = 131 M( )(1) s(-)(1)). These results demonstrate that S'-P' interactions can be utilized to increase the efficacy and selectivity of (acyloxy)methyl ketone inactivators. PMID- 10828966 TI - Downregulation of atrial natriuretic peptide ANP-C receptor is associated with alterations in G-protein expression in A10 smooth muscle cells. AB - Atrial natriuretic peptide (ANP) receptors A and B are guanylyl cyclase receptors, whereas ANP-C receptors are coupled to adenylyl cyclase through inhibitory guanine nucleotide (Gi) protein. ANP has been shown to downregulate ANP-A and -B receptors and cGMP response in various tissues. In the present studies, we have examined the regulation of ANP-C receptor-adenylyl cyclase signal transduction by ANP and [des(Gln(18),Ser(19),Gln(20),Leu(21), Gly(22))ANP(4-23)-NH(2)](C-ANP(4-23)) that interacts specifically with ANP-C receptor in A10 smooth muscle cells (SMC). Treatment of the cells with C-ANP(4 23) for 24 h resulted in a reduction in ANP receptor binding activity. [(125)I]ANP(99-126) bound to control and C-ANP(4-23)-treated cell membranes at a single site with dissociation constants of 33.7 +/- 6 and 35.0 +/- 4.5 pM and B(max) of 74.0 +/- 5.0 and 57.6 +/- 4.0 fmol/mg of protein, respectively. C-ANP(4 23) inhibited adenylyl cyclase activity in a concentration-dependent manner in control cells. A maximal inhibition observed was about 30-40% with an apparent K(i) of about 1 nM; however, this inhibition was completely attenuated in cells pretreated with ANP(99-126) or C-ANP(4-23) (10(-7) M). However, the inhibition of adenylyl cyclase by 17-amino acid peptide (RRNHQEESNIGKHRELR) (R17A) of cytoplasmic domain of ANP-C receptor was attenuated by about 50% but was not completely abolished by C-ANP(4-23) treatment. The attenuation of C-ANP(4-23) mediated inhibition of adenylyl cyclase was dependent on the concentration and time of pretreatment of the cells with C-ANP(4-23). In addition, angiotensin II- (Ang II-) mediated inhibition of adenylyl cyclase ( approximately 30%) was also abolished by C-ANP(4-23) treatment, indicating that the desensitization elicited by ANP was heterologous. In addition, C-ANP(4-23) treatment decreased the expression of Gialpha-2 and Gialpha-3 proteins by about 40 and 60%, respectively, and their mRNA by 40%. However, the levels of Gi proteins were not altered when the cells were treated for shorter period of time (2-4 h) or with lower concentrations of C-ANP(4-23) (10(-10) M). On the other hand, the levels of Gsalpha but not of Gbeta were increased by about 35% by C-ANP(4-23) treatment. Furthermore, the stimulations exerted by GTPgammaS, isoproterenol, FSK, and NaF on adenylyl cyclase were also augmented in cells treated with C-ANP(4-23). These results indicate that C-ANP(4-23) treatment of A10 cells desensitizes ANP-C receptor-mediated inhibition of adenylyl cyclase which may be due to the downregulation of ANP-C receptor and decreased expression of Gialpha proteins to which these receptors are coupled. PMID- 10828967 TI - Role of asparagine-linked oligosaccharides in the function of the rat PTH/PTHrP receptor. AB - The receptor for parathyroid hormone (PTH) and PTH-related peptide (PTHrP) is a G protein-coupled receptor with four potential sites for N-linked glycosylation. The contribution of the oligosaccharide moieties to cell surface expression, ligand binding, and signal transduction was investigated. Site-directed mutagenesis of the rat PTH/PTHrP receptor cDNA was performed at single or combination of the four potential glycosylation sites to determine the effect of the putative carbohydrate chains on the activities of the receptor. The results revealed that all four potential N-glycosylation sites in the PTH/PTHrP receptor are glycosylated. Receptors missing a single or multiple glycosylation consensus but with at least one intact glycosylation site expressed sufficiently and functioned normally. In contrast, the nonglycosylated receptor, in which all four glycosylation sites were mutated, is deficient in these functions. These data indicate important roles for N-linked glycosylation in PTH/PTHrP receptor functions. PMID- 10828968 TI - Analysis of the role of interfacial tryptophan residues in controlling the topology of membrane proteins. AB - Tryptophans have a high affinity for the membrane-water interface and have been suggested to play a role in determining the topology of membrane proteins. We investigated this potential role experimentally, using mutants of the single spanning Pf3 coat protein, whose transmembrane topologies are sensitive to small changes in amino acid sequence. Mutants were constructed with varying numbers of tryptophans flanking the transmembrane region and translocation was assessed by an in vitro translation/translocation system. Translocation into Escherichia coli inner membrane vesicles could take place under a variety of experimental conditions, with co- or posttranslational assays and proton motive force dependent or -independent mutants. It was found that translocation can even occur in pure lipid vesicles, under which conditions the tryptophans must directly interact with the lipids. However, under all these conditions tryptophans neither inhibited nor stimulated translocation, demonstrating that they do not affect topology and suggesting that this may be universal for tryptophans in membrane proteins. In contrast, we could demonstrate that lysines clearly prefer to stay on the cis-side of the membrane, in agreement with the positive-inside rule. A statistical analysis focusing on interfacially localized residues showed that in single-spanning membrane proteins lysines are indeed located on the inside, while tryptophans are preferentially localized at the outer interface. Since our experimental results show that the latter is not due to a topology-determining role, we propose instead that tryptophans fulfill a functional role as interfacially anchoring residues on the trans-side of the membrane. PMID- 10828969 TI - Self-aggregation of surfactant protein A. AB - Environmental factors of physiological relevance such as pH, calcium, ionic strength, and temperature can affect the state of self-aggregation of surfactant protein A (SP-A). We have studied the secondary structure of different SP-A aggregates and analyzed their fluorescence characteristics. (a) We found that self-aggregation of SP-A can be Ca(2+)-dependent. The concentration of Ca(2+) needed for half-maximal self-association (K(a)(Ca)()2+) depended on the presence of salts. Thus, at low ionic strength, K(a)(Ca)()2+ was 2.3 mM, whereas at physiological ionic strength, K(a)(Ca)()2+ was 2.35 microM. Circular dichroism and fluorescence measurements of Ca(2+)-dependent SP-A aggregates indicated that those protein aggregates formed in the absence of NaCl are structurally different from those formed in its presence. (b) We found that self-aggregation of SP-A can be pH-dependent. Self-aggregation of SP-A induced by H(+) was highly influenced by the presence of salts, which reduced the extent of self-association of the protein. The presence of both salts and Ca(2+) attenuated even more the effects of acidic media on SP-A self-aggregation. (c) We found that self-aggregation of SP-A can be temperature-dependent. At 20 degrees C, SP-A underwent self aggregation at physiological but not at low ionic strength, in the presence of EDTA. All of these aggregates were dissociated by either adding EDTA (a), increasing the pH to neutral pH (b), or increasing the temperature to 37 degrees C (c). Dissociation of Ca(2+)-induced protein aggregates at low ionic strength was accompanied by an irreversible loss of both SP-A secondary structure and SP-A dependent lipid aggregation properties. On the other hand, temperature-dependent experiments indicated that a structurally intact collagen-like domain was required for either Ca(2+)- or Ca(2+)/Na(+)-induced SP-A self-aggregation but not for H(+)-induced protein aggregation. PMID- 10828970 TI - Enzyme-induced strain/distortion in the ground-state ES complex in beta-lactamase catalysis revealed by FTIR. AB - Class A beta-lactamases hydrolyze penicillins and other beta-lactams via an acyl enzyme catalytic mechanism. Ser70 is the active site nucleophile. By constructing the S70A mutant, which is unable to form the acyl-enzyme intermediate, it was possible to make stable ES complexes with various substrates. The stability of such Michaelis complexes permitted acquisition of their infrared spectra. Comparison of the beta-lactam carbonyl stretch frequency (nu(CO)) in the free and enzyme-bound substrate revealed an average decrease of 13 cm(-)(1), indicating substantial strain/distortion of the lactam carbonyl when bound in the ES complex. Interestingly, regardless of the frequency of the C=O stretch in the free substrate, when complexed to Bacillus licheniformis beta-lactamase, the frequency was always 1755 +/- 2 cm(-)(1). This suggests the active site environment induces a similar conformation of the beta-lactam in all substrates when bound to the enzyme. Using deuterium substitution, it was shown that the "oxyanion hole", which involves hydrogen bonding to two backbone amides, is the major source of the enzyme-induced strain/distortion. The very weak catalytic activity of the S70A beta-lactamase suggests enzyme-facilitated hydrolysis due to substrate distortion on binding to the enzyme. Thus the binding of the substrate in the active site induces substantial strain and distortion that contribute significantly to the overall rate enhancement in beta-lactamase catalysis. PMID- 10828971 TI - Escherichia coli dimethylallyl diphosphate:tRNA dimethylallyltransferase: essential elements for recognition of tRNA substrates within the anticodon stem loop. AB - Escherichia coli dimethylallyl diphosphate:tRNA dimethylallyltransferase (DMAPP tRNA transferase) catalyzes the alkylation of the exocyclic amine of A37 by a dimethylallyl unit in tRNAs with an adenosine in the third anticodon position (position 36). By use of purified recombinant enzyme, steady- state kinetic studies were conducted with chemically synthesized RNA oligoribonucleotides to determine the essential elements within the tRNA anticodon stem-loop structure required for recognition by the enzyme. A 17-base oligoribonucleotide corresponding to the anticodon stem-loop of E. coli tRNA(Phe) formed a stem-loop minihelix (minihelix(Phe)) when annealed rapidly on ice, while the same molecule formed a duplex structure with a central loop when annealed slowly at higher concentrations. Both the minihelix and duplex structures gave k(cat)s similar to that for the normal substrate (full-length tRNA(Phe) unmodified at A37), although the K(m) for minihelix(Phe) was approximately 180-fold higher than full-length tRNA. The A36-A37-A38 motif, which is completely conserved in tRNAs modified by the enzyme, was found to be important for modification. Changing A36 to G in the minihelix resulted in a 260-fold reduction in k(cat) compared to minihelix(Phe) and a 13-fold increase in K(m). An A38G variant was modified with a 9-fold reduction in k(cat) and a 5-fold increase in K(m). A random coil 17-base oligoribonucleotide in which the loop sequence of E. coli tRNA(Phe) was preserved, but the 5 base pair helix stem was completely disrupted and showed no measurable activity, indicating that a helix-loop structure is essential for recognition. Finally, altering the identity of several base pairs in the helical stem did not have a major effect on catalytic efficiency, suggesting that the enzyme does not make base-specific contacts important for binding or catalysis in this region. PMID- 10828972 TI - Multiple posttranslational modifications at distinct sites contribute to heterogeneity of the lipoprotein cytochrome bo(3). AB - The heme-copper cytochrome oxidase of Escherichia coli (cytochrome bo(3)) was tagged with oligohistidine at the C-terminus of the small noncatalytic subunit IV. After detergent solubilization, the enzyme was purified by a one-step procedure with immobilized metal affinity chromatography. Using different cytochrome bo(3) constructs as reference, the products were investigated by mass spectroscopical and immunological methods. Several posttranslational modifications of subunits II, III, and IV were observed: (1) N-terminal methionines of subunits III and IV are split off. (2) Fifty percent of subunit III polypeptides are acetylated, presumably at the N-terminal alanine. (3) Lipoprotein processing of subunit II involves cleavage of the signal peptide. (4) Maturation of subunit II [Ma, J., Katsonouri, A., and Gennis, R. B. (1997) Biochemistry 36, 11298-11303] alters the structure of the N-terminal cysteine by N-palmitoylation and S-glyceryldipalmitoylation. (5) A hexapeptide is split off from the C-terminus of subunit II. This happens subsequently to the N-terminal lipoprotein processing step and is dependent on the growth state of cells. PMID- 10828973 TI - Role of side-chains in the cooperative beta-hairpin folding of the short C terminal fragment derived from streptococcal protein G. AB - A short C-terminal fragment of immunoglobulin-binding domain of streptococcal protein G is known to form nativelike beta-hairpin at physiological conditions. To understand the cooperative folding of the short peptide, eight Ala-substituted mutants of the fragment were investigated with respect to their structural stabilities by analyzing temperature dependence of NMR signals. On comparison of the obtained thermodynamic parameters, we found that the nonpolar residues Tyr45 and Phe52 and the polar residues Asp46 and Thr49 are crucial for the beta-hairpin folding. The results suggest a strong interaction between the nonpolar side chains that participates in a putative hydrophobic cluster and that the polar side chains form a fairly rigid conformation around the loop (46-51). We also investigated the complex formation of the mutants with N-terminal fragment at the variety of temperature to get their thermal unfolding profiles and found that the mutations on the residues Asp46 and Thr49 largely destabilized the complexes, while substitution of Asp47 slightly stabilized the complex. From these results, we deduced that both the hydrophobic cluster formation and the rigidity of the loop (46-51) cooperatively stabilize the beta-hairpin structure of the fragment. These interactions which form a stable beta-hairpin may be the initial structural scaffold which is important in the early folding events of the whole domain. PMID- 10828974 TI - Val(659)-->Glu mutation within the transmembrane domain of ErbB-2: effects measured by (2)H NMR in fluid phospholipid bilayers. AB - Certain point mutations within the hydrophobic transmembrane domains of class I receptor tyrosine kinases have been associated with oncogenic transformation in vitro and in vivo [Gullick, J., and Srinivasan, R. (1998) Breast Cancer Res. Treat. 52, 43-53]. An important example is the replacement of a single (hydrophobic) valine by (charged) glutamate in the rat protein, Neu, and in the homologous human protein, ErbB-2. It has been suggested that the oncogenic nature of this Val-->Glu substitution may derive from alteration of the transmembrane domain's ability to take part in direct side-to-side associations. In the present work, we examined the basis of this phenomenon by studying transmembrane portions of ErbB-2 in fluid bilayer membranes. An expression system was designed to produce such peptides from the wild-type ErbB-2, and from an identical region of the transforming mutant in which Val(659) is replaced by Glu. All peptides were 50-mers, containing the appropriate transmembrane domain plus contiguous stretches of amino acids from the cytoplasmic and extracellular domains. Deuterium heteronuclear probes were incorporated into alanine side chains (thus, each alanine -CH(3) side chain became -CD(3)). Given the presence of natural alanine residues at positions 648 and 657 within ErbB-2, this approach afforded heteronuclear probes within the motif Ser(656)AlaValValGlu(660), thought to be important for homodimer formation, and nine residues upstream of this site. Further peptides were produced, by site-directed mutagenesis, to confirm spectral assignments and to provide an additional probe location at position 670 (11 residues downstream of the motif region). On SDS-polyacrylamide gels, the transmembrane peptides migrated as predominant monomers in equilibrium with smaller populations of homodimers/-oligomers. CD spectra of both wild-type and transforming mutant peptides were consistent with the transmembrane portions being basically alpha-helical. (2)H NMR spectra of each transmembrane peptide were obtained in fluid phospholipid bilayers of 1-palmitoyl-2 oleoylphosphatidylcholine (POPC) from 35 to 65 degrees C. Results were consistent with the concept that the glutamic acid residue characterizing the mutant is uncharged at neutral pH. Narrowed spectral components from species rotating rapidly and symmetrically within the membrane appeared to represent monomeric peptide. Mutation of Val(659) to Glu within the hydrophobic domain induced changes in side chain angulation of at least 6-8 degrees at Ala(657) (i.e., within the five amino acid motif thought to be involved in homodimer formation), and downstream of this site to residue 670. There was little evidence of effect at the upstream site (Ala(648)) at the membrane surface. This result argues that the transforming mutation is associated with significant intramolecular rearrangement of the monomeric transmembrane helix-extending over some four helix turns-which could influence its lateral associations. In addition, temperature effects on spectral quadrupole splittings suggested that there is greater peptide backbone flexibility for the wild-type transmembrane region. PMID- 10828975 TI - Oblique membrane insertion of viral fusion peptide probed by neutron diffraction. AB - Fusion peptides mimic the membrane fusion activities of the larger viral proteins from which they derive their sequences. A possible mode of activity involves their oblique insertion into lipid bilayers, causing membrane disruption by promoting highly curved hemifusion intermediates, leading to fusion. We have determined the location and orientation of the simian immunodeficiency virus (SIV) fusion peptide in planar lipid bilayers using neutron lamellar diffraction. The helical axis of the peptide adopts an angle of 55 degrees relative to the membrane normal, while it positions itself nearest the lipid bilayer surface. This is the first direct observation of the structural interaction between a fusion peptide and a phospholipid bilayer. PMID- 10828976 TI - Analysis of a water mediated protein-protein interactions within RNase T1. AB - Buried and well-ordered solvent molecules are an integral part of each folded protein. For a few individual water molecules, the exchange kinetics with solvent have been described in great detail. So far, little is known about the energetics of this exchange process. Here, we present an experimental approach to investigate water-mediated intramolecular protein-protein interactions by use of double mutant cycles. As a first example, we analyzed the interdependence of the contribution of two side chains (Asn9 and Thr93) to the conformational stability of RNase T1. In the folded state, both side chains are involved in the "solvation of the same water molecule WAT1. The coupling of the contributions of Asn9 and Thr93 to the conformational stability of RNase T1 was measured by urea unfolding and differential scanning calorimetry. The structural integrity of each mutant was analyzed by X-ray crystallography. We find that the effects of the Asn9Ala and the Thr93Ala mutations on the conformational stability are additive in the corresponding double mutant. We conclude that the free energy of the WAT1 mediated intramolecular protein-protein interaction in the folded state is very similar to solvent mediated protein-protein interaction in the unfolded state. PMID- 10828977 TI - Pyrene excimer fluorescence: a spatially sensitive probe to monitor lipid-induced helical rearrangement of apolipophorin III. AB - Manduca sexta apolipophorin III (apoLp-III), an 18-kDa, monomeric, insect hemolymph apolipoprotein, is comprised of five amphipathic alpha-helices arranged as a globular bundle in the lipid-free state. Upon lipid binding, it is postulated that the bundle opens, exposing a continuous hydrophobic surface which becomes available for lipid interaction. To investigate lipid binding-induced helical rearrangements, we exploited the unique fluorescence characteristics of N (1-pyrene)maleimide. Pyrene is a spatially sensitive extrinsic fluorescent probe, which forms excited-state dimers (excimers) upon close encounter with another pyrene molecule. Cysteine residues were introduced into apoLp-III (which otherwise lacks cysteine) at Asn 40 (helix 2) and/or Leu 90 (helix 3), creating two single-cysteine mutants (N40C-apoLp-III and L90C-apoLp-III) and N40C/L90C apoLp-III, a double-cysteine mutant, which were labeled with pyrene maleimide. Pyrene-labeled N40C/L90C-apoLp-III, but not the pyrene-labeled single-cysteine mutants, exhibited strong excimer fluorescence in the lipid-free, monomeric state. Guanidine hydrochloride titration and temperature studies revealed a loss in excimer fluorescence, accompanied by a loss in the molar ellipticity of the protein. When apoLp-III interacts with phospholipid vesicles to form disklike complexes, a significant loss in excimer fluorescence was noted, indicating that the helices bearing the pyrene moieties diverge from each other. Pyrene excimer fluorescence was further employed to examine the relative orientation of lipid bound apoLp-III molecules. Pyrene-labeled N40C- or L90C-apoLp-III displayed no excimer fluorescence in the disk complexes, while complexes prepared with an equal mixture of both single-labeled mutants did emit excimer fluorescence, indicating apoLp-III adopts a preferred nonrandom orientation around the perimeter of the bilayer disk. These studies establish pyrene excimer fluorescence as a useful spectroscopic tool to address intra- and intermolecular interactions of exchangeable apolipoproteins upon binding to lipid. PMID- 10828978 TI - AhpF can be dissected into two functional units: tandem repeats of two thioredoxin-like folds in the N-terminus mediate electron transfer from the thioredoxin reductase-like C-terminus to AhpC. AB - AhpF, the flavin-containing component of the Salmonella typhimurium alkyl hydroperoxide reductase system, catalyzes the NADH-dependent reduction of an active-site disulfide bond in the other component, AhpC, which in turn reduces hydroperoxide substrates. The amino acid sequence of the C-terminus of AhpF is 35% identical to that of thioredoxin reductase (TrR) from Escherichia coli. AhpF contains an additional 200-residue N-terminal domain possessing a second redox active disulfide center also required for AhpC reduction. Our studies indicate that this N-terminus contains a tandem repeat of two thioredoxin (Tr)-like folds, the second of which contains the disulfide redox center. Structural and catalytic properties of independently expressed fragments of AhpF corresponding to the TrR like C-terminus (F[208-521]) and the 2Tr-like N-terminal domain (F[1-202]) have been addressed. Enzymatic assays, reductive titrations, and circular dichroism studies of the fragments indicate that each folds properly and retains many functional properties. Electron transfer between F[208-521] and F[1-202] is, however, relatively slow (4 x 10(4) M(-)(1) s(-)(1) at 25 degrees C) and nonsaturable up to 100 microM F[1-202]. TrR is nearly as efficient at F[1-202] reduction as is F[208-521], although neither the latter fragment, nor intact AhpF, can reduce Tr. An engineered mutant AhpC substrate with a fluorophore attached via a disulfide bond has been used to demonstrate that only F[1-202], and not F[208-521], is capable of electron transfer to AhpC, thereby establishing the direct role this N-terminal domain plays in mediating electron transfer between the TrR-like part of AhpF and AhpC. PMID- 10828979 TI - Peroxidase activity in prostaglandin endoperoxide H synthase-1 occurs with a neutral histidine proximal heme ligand. AB - Prostaglandin endoperoxide H synthases-1 and -2 (PGHS-1 and -2) convert arachidonic acid to prostaglandin H(2) (PGH(2)), the committed step in prostaglandin and thromboxane formation. Interaction of peroxides with the heme sites in PGHSs generates a tyrosyl radical that catalyzes subsequent cyclooxygenase chemistry. To study the peroxidase reaction of ovine oPGHS-1, we combined spectroscopic and directed mutagenesis data with X-ray crystallographic refinement of the heme site. Optical and Raman spectroscopy of oxidized oPGHS-1 indicate that its heme iron (Fe(3+)) exists exclusively as a high-spin, six coordinate species in the holoenzyme and in heme-reconstituted apoenzyme. The sixth ligand is most likely water. The cyanide complex of oxidized oPGHS-1 has a six-coordinate, low-spin ferric iron with a v[Fe-CN] frequency at 445 cm(-)(1); a monotonic sensitivity to cyanide isotopomers that indicates the Fe-CN adduct has a linear geometry. The ferrous iron in reduced oPGHS-1 adopts a high-spin, five coordinate state that is converted to a six-coordinate, low-spin geometry by CO. The low-frequency Raman spectrum of reduced oPGHS-1 reveals two v[Fe-His] frequencies at 206 and 222 cm(-)(1). These vibrations, which disappear upon addition of CO, are consistent with a neutral histidine (His388) as the proximal heme ligand. The refined crystal structure shows that there is a water molecule located between His388 and Tyr504 that can hydrogen bond to both residues. However, substitution of Tyr504 with alanine yields a mutant having 46% of the peroxidase activity of native oPGHS-1, establishing that bonding of Tyr504 to this water is not critical for catalysis. Collectively, our results show that the proximal histidine ligand in oPGHS-1 is electrostatically neutral. Thus, in contrast to most other peroxidases, a strongly basic proximal ligand is not necessary for peroxidase catalysis by oPGHS-1. PMID- 10828980 TI - Crystal structure and iron-binding properties of the R210K mutant of the N-lobe of human lactoferrin: implications for iron release from transferrins. AB - Lactoferrin (Lf) and serum transferrin (Tf) combine high-affinity iron binding with an ability to release this iron at reduced pH. Lf, however, retains iron to significantly lower pH than Tf, giving the two proteins distinct functional roles. In this paper, we compared the iron-release profiles for human Lf, Tf, and their N-lobe half-molecules Lf(N) and Tf(N) and showed that half of the difference in iron retention at low pH ( approximately 1.3 pH units) results from interlobe interactions in Lf. To probe factors intrinsic to the N-lobes, we further examined the specific role of two basic residues that are proposed to form a pH-sensitive dilysine trigger for iron release in the N-lobe of Tf [Dewan, J. C., Mikami, B., Hirose, M., and Sacchettini, J. C. (1993) Biochemistry 32, 11963-11968] by mutating Arg 210 to Lys in the N-lobe half-molecule Lf(N). The R210K mutant was expressed, purified, and crystallized, and its crystal structure was determined and refined at 2.0-A resolution to a final R factor (R(free)) of 19.8% (25.0%). The structure showed that Lys 210 and Lys 301 in R210K do not form a dilysine interaction like that between Lys 206 and Lys 296 in human Tf. The R210K mutant retained iron to lower pH than Tf(N), consistent with the absence of the dilysine interaction but released iron at approximately 0.7 pH units higher than Lf(N). We conclude that (i) the ability of Lf to retain iron to significantly lower pH than Tf is due equally to interlobe interactions and to the absence in Lfs of an interaction analogous to the dilysine pair in Tfs, even when two lysines are present at the corresponding sequence positions, and (ii) an appropriately positioned basic residue (Arg 210 in human Lf) modulates iron release by inhibiting protonation of the N-lobe iron ligands, specifically His 253. PMID- 10828981 TI - Backbone dynamics of escherichia coli adenylate kinase at the extreme stages of the catalytic cycle studied by (15)N NMR relaxation. AB - Adenylate kinase from Escherichia coli (AKeco), consisting of a single 23.6 kDa polypeptide chain folded into domains CORE, AMPbd, and LID, catalyzes the reaction AMP + ATP --> 2ADP. Domains LID and AMPbd execute large-scale movements during catalysis. Backbone dynamics of ligand-free and AP(5)A-inhibitor-bound AKeco were studied comparatively with (15)N NMR relaxation methods. Overall diffusion with correlation times of 15.05 (11.42) ns and anisotropy D(parallel)/D(perp) = 1.25 (1.10), and fast internal motions with correlation times up to 100 ps (50 ps), were determined for AKeco (AKecoAP(5)A). Fast internal motions affect 93% of the AKeco sites, with pronounced preference for domains AMPbd and LID, and 47% of the AKecoAP(5)A sites, with limited variability along the chain. The mean squared generalized order parameters, , of secondary structure elements and loops are affected by ligand binding differentially and in a domain-specific manner. Nanosecond motions predominate within AMPbd. Prominent exchange contributions, associated in particular with residue G10 of the nucleotide-binding P-loop motif, are interpreted to reflect hydrogen-bond dynamics at the inhibitor-binding site. The hypothesis of energetic counter balancing of substrate binding based on crystallographic data is strongly supported by the solution NMR results. Correlations between backbone dynamics and domain displacement are established. PMID- 10828982 TI - Identification of the amino acid residues of the amino terminus of vimentin responsible for DNA binding by enzymatic and chemical sequencing and analysis by MALDI-TOF. AB - The amino acid residues responsible for stable binding of nucleic acids by the intermediate filament (IF) subunit protein vimentin were identified by a combination of enyzmatic and chemical ladder sequencing of photo-cross-linked vimentin-oligodeoxyribonucleotide complexes and analysis by MALDI-TOF mass spectrometry. Three tryptic peptides of vimentin (vim(28)(-)(35), vim(36)(-)(49), and vim(50)(-)(63)) were found to be cross-linked to oligo(dG.BrdU)(12). dG.3' FITC. From a methodological standpoint, it was necessary to remove the bulk of the bound oligonucleotide by digestion with nuclease P1 to get reproducible spectra for most of the peptides studied. Additionally, removal of the phosphate group of the residually bound dUMP or modification of the amino terminus of the peptide-oligonucleotide complexes with dimethylaminoazobenzene isothiocyanate dramatically improved the quality of the MALDI-TOF spectra obtained, particularly for the vim(28)(-)(35) peptide. A single Tyr residue within each of these peptides (Tyr(29), Tyr(37), and Tyr(52)) was unequivocally demonstrated to be the unique site of cross-linking in each peptide. These three Tyr residues are contained within the two beta-ladder DNA-binding wings proposed for the middle of the vimentin non-alpha-helical head domain. The experimental approach described should be generally applicable to the study of protein-nucleic acid interactions and is currently being employed to characterize the DNA-binding sites of several other IF subunit proteins. PMID- 10828983 TI - A thioredoxin from the hyperthermophilic archaeon Methanococcus jannaschii has a glutaredoxin-like fold but thioredoxin-like activities. AB - A thioredoxin homologue (Mj0307) from the hyperthermophilic archaeon Methanococcus jannaschii (MjTRX) was cloned, produced in E. coli, and compared to the thioredoxin from E. coli (ETRX). The secondary structure profile of MjTRX obtained by NMR spectroscopy shows that it has four beta-sheets and three alpha helices arranged in betaalphabetaalphabetabetaalpha, similar to that of glutaredoxin. However, MjTRX supports the growth of T7 bacteriophage in E. coli and is weakly reduced by the thioredoxin reductase from E. coli, indicating that MjTRX is functionally closer to a thioredoxin than a glutaredoxin. MjTRX has higher specific insulin reductase activity than ETRX and retained its full activity over 4 days at 95 degrees C, whereas ETRX lost its activity in 150 min. The standard state redox potential of MjTRX is about -277 mV, which is the lowest value thus far known among redox potentials of the thioredoxin superfamily. This indicates that the lower redox potential is necessary in keeping catalytic disulfide bonds reduced in the cytoplasm and in coping with oxidative stress in an anaerobic hyperthermophile. PMID- 10828984 TI - Ceramide interaction with the respiratory chain of heart mitochondria. AB - A study is presented on the interaction of ceramide with the respiratory chain of rat heart mitochondria, and a comparison is made between the effects elicited by short- and long-chain ceramides. N-Acetylsphingosine (C(2)-ceramide) and N palmitoylsphingosine (C(16)-ceramide) inhibited to the same extent the pyruvate+malate-dependent oxygen consumption. Succinate-supported respiration was also inhibited by ceramides, but this activity was substantially restored upon the addition of cytochrome c, which, on the contrary, was ineffective toward the ceramide-inhibited NADH-linked substrate oxidation. Direct measurements showed that short- and long-chain ceramides caused a large release of cytochrome c from mitochondria. The ceramide-dependent inhibition of pyruvate+malate and succinate oxidation caused reactive oxygen species to be produced at the level of either complex I or complex III. The activity of the cytochrome c oxidase, measured as ascorbate/TMPD oxidase activity, was significantly stimulated and inhibited by C(2)- and C(16)-ceramide, respectively. Similar effects were observed on the activity of the individual respiratory complexes isolated from bovine heart. Short- and long-chain ceramides had definitely different effects on the mitochondrial membrane potential. C(2)-ceramide caused an almost complete collapse of the respiration-dependent membrane potential, whereas C(16)-ceramide had a negligible effect. Similar results were obtained when the potential was generated in liposome-reconstituted complex III respiring at the steady-state. Furthermore, C(2)-ceramide caused a drop of the membrane potential generated by ATP hydrolysis instead of respiration, whereas C(16)-ceramide did not. Finally, only short-chain ceramides inhibited markedly the reactive oxygen species generation associated with membrane potential-dependent reverse electron flow from succinate to complex I. The emerging indication is that the short-chain ceramide-dependent collapse of membrane potential is a consequence of their ability to perturb the membrane structure, leading to an unspecific increase of its permeability. PMID- 10828985 TI - Resonance raman studies of oxo intermediates in the reaction of pulsed cytochrome bo with hydrogen peroxide. AB - Cytochrome bo from Escherichia coli, a member of the heme-copper terminal oxidase superfamily, physiologically catalyzes reduction of O(2) by quinols and simultaneously translocates protons across the cytoplasmic membrane. The reaction of its ferric pulsed form with hydrogen peroxide was investigated with steady state resonance Raman spectroscopy using a homemade microcirculating system. Three oxygen-isotope-sensitive Raman bands were observed at 805/X, 783/753, and (767)/730 cm(-)(1) for intermediates derived from H(2)(16)O(2)/H(2)(18)O(2). The experiments using H(2)(16)O(18)O yielded no new bands, indicating that all the bands arose from the Fe=O stretching (nu(Fe)(=)(O)) mode. Among them, the intensity of the 805/X cm(-)(1) pair increased at higher pH, and the species giving rise to this band seemed to correspond to the P intermediate of bovine cytochrome c oxidase (CcO) on the basis of the reported fact that the P intermediate of cytochrome bo appeared prior to the formation of the F species at higher pH. For this intermediate, a Raman band assignable to the C-O stretching mode of a tyrosyl radical was deduced at 1489 cm(-)(1) from difference spectra. This suggests that the P intermediate of cytochrome bo contains an Fe(IV)=O heme and a tyrosyl radical like compound I of prostaglandin H synthase. The 783/753 cm(-)(1) pair, which was dominant at neutral pH and close to the nu(Fe)(=)(O) frequency of the oxoferryl intermediate of CcO, presumably arises from the F intermediate. On the contrary, the (767)/730 cm(-)(1) species has no counterpart in CcO. Its presence may support the branched reaction scheme proposed previously for O(2) reduction by cytochrome bo. PMID- 10828986 TI - Differential effects of chilling-induced photooxidation on the redox regulation of photosynthetic enzymes. AB - Photosynthesis in plant species that are evolutionarily adapted for growth in warm climates is highly sensitive to illumination under cool conditions. Although it is well documented that illumination of these sensitive species under cool conditions results in the photosynthetic production of reactive oxygen molecules, the underlying mechanism for the inhibition of photosynthesis remains uncertain. Determinations of chloroplast fructose-1,6-bisphosphatase and sedoheptulose-1,7 bisphosphatase activity showed that the light-dependent, reductive activation of these key carbon reduction cycle enzymes was substantially inhibited in tomato (Lycopersicon esculentum) following illumination at 4 degrees C. However, other chloroplast enzymes also dependent on thioredoxin-mediated reductive activation were largely unaffected. We performed equilibrium redox titrations to investigate the thermodynamics of the thiol/disulfide exchange between thioredoxin f and the regulatory sulfhydryl groups of fructose-1,6-bisphosphatase, sedoheptulose-1,7 bisphosphatase, phosphoribulokinase, NADP-glyceraldehyde phosphate dehydrogenase, and the chloroplast ATPsynthase. We determined that the redox midpoint potentials for the regulatory sulfhydryl groups of the various enzymes spanned a broad range ( approximately 50 mV at pH 7. 9). The electron-sharing equilibria among thioredoxin f and its target enzymes largely explained the differential effects of photooxidation induced at low temperature on thioredoxin-mediated activation of chloroplast enzymes in tomato. These results not only provide a plausible mechanism for the low-temperature-induced inhibition of photosynthesis in this important group of plants, but also provide a quantitative basis to evaluate the influence of thioredoxin/target enzyme electron-sharing equilibria on the differential activation and deactivation kinetics of thioredoxin-regulated chloroplast enzymes. PMID- 10828987 TI - Scavenging of peroxynitrite by oxyhemoglobin and identification of modified globin residues. AB - Peroxynitrite is a strong oxidant involved in cell injury. In tissues, most of peroxynitrite reacts preferentially with CO(2) or hemoproteins, and these reactions affect its fate and toxicity. CO(2) promotes tyrosine nitration but reduces the lifetime of peroxynitrite, preventing, at least in part, membrane crossing. The role of hemoproteins is not easily predictable, because the heme intercepts peroxynitrite, but its oxidation to ferryl species and tyrosyl radical(s) may catalyze tyrosine nitration. The modifications induced by peroxynitrite/CO(2) on oxyhemoglobin were determined by mass spectrometry, and we found that alphaTyr42, betaTyr130, and, to a lesser extent, alphaTyr24 were nitrated. The suggested nitration mechanism is tyrosyl radical formation by long range electron transfer to ferrylhemoglobin followed by a reaction with (*)NO(2). Dityrosine (alpha24-alpha42) and disulfides (beta93-beta93 and alpha104-alpha104) were also detected, but these cross-linkings were largely due to modifications occurring under the denaturing conditions employed for mass spectrometry. Moreover, immunoelectrophoretic techniques showed that the 3-nitrotyrosine content of oxyhemoglobin sharply increased only in molar excess of peroxynitrite, thus suggesting that this hemoprotein is not a catalyst of nitration. The noncatalytic role may be due to the formation of the nitrating species (*)NO(2) mainly in molar excess of peroxynitrite. In agreement with this hypothesis, oxyhemoglobin strongly inhibited tyrosine nitration of a target dipeptide (Ala Tyr) and of membrane proteins from ghosts resealed with oxyhemoglobin. Erythrocytes were poor inhibitors of Ala-Tyr nitration on account of the membrane barrier. However, at the physiologic hematocrit, Ala-Tyr nitration was reduced by 65%. This "sink" function was facilitated by the huge amount of band 3 anion exchanger on the cell membrane. We conclude that in blood oxyhemoglobin is a peroxynitrite scavenger of physiologic relevance. PMID- 10828988 TI - Interactions of human fibrinogens with factor XIII: roles of calcium and the gamma' peptide. AB - Plasma factor XIII is the zymogen of the transglutaminase factor XIIIa. This enzyme catalyzes the formation of isopeptide cross-links between fibrin molecules in nascent blood clots that greatly increase the mechanical stability of clots and their resistance to thrombolytic enzymes. We have characterized the solution interactions of factor XIII with two variants of fibrinogen, the soluble precursor of fibrin. Both the predominant fibrinogen gamma(A)/gamma(A) and the major variant gamma(A)/gamma' form complexes with a 2 fibrinogen:1 factor XIII ratio. The absence of detectable concentrations of 1:1 complexes in equilibrium mixtures containing free factor XIII and 2:1 complexes suggests that this interaction is cooperative. Factor XIII binds fibrinogen gamma(A)/gamma' approximately 20-fold more tightly than fibrinogen gamma(A)/gamma(A), and the interaction with fibrinogen gamma(A)/gamma' (but not fibrinogen gamma(A)/gamma(A)) is accompanied by a significant release of Ca(2+). Taken together, these results suggest that the strikingly anionic gamma' C-terminal sequence contains features that are important for factor XIII binding. Consistent with this notion, a synthetic 20-residue polypeptide containing the gamma' sequence was found to associate with factor XIII in a 2:1 molar ratio and act as an efficient competitor for fibrinogen gamma(A)/gamma' binding. PMID- 10828989 TI - A quantitative study of the interactions of Bacillus anthracis edema factor and lethal factor with activated protective antigen. AB - Bacillus anthracis secretes three proteins, which associate in binary combinations to form toxic complexes at the surface of mammalian cells. Receptor bound protective antigen (PA) is proteolytically activated, yielding a 63 kDa fragment (PA(63)). PA(63) oligomerizes into heptamers, which bind edema factor (EF) or lethal factor (LF) to form the toxic complexes. We undertook a quantitative analysis of the interactions of EF with PA(63) by means of surface plasmon resonance (SPR) measurements. Heptameric PA(63) was covalently bound by amine coupling to an SPR chip, or noncovalently bound via a C-terminal hexahistidine tag on the protein to Ni(2+)nitrilotriacetate groups on the chip. Values of k(on) and k(off) for EF at 23 degrees C were approximately 3 x 10(5) M( )(1) s(-)(1) and (3-5) x 10(-)(4) s(-)(1), respectively, giving a calculated K(d) of (1-2) x 10(-)(9) M. A similar value of K(d) (7 x 10(-)(10) M) was obtained when we measured the binding of radiolabeled EF to receptor-bound PA(63) on the surface of L6 cells (at 4 degrees C). Each of these analyses was also performed with LF and LF(N) (the N-terminal 255 residues of LF), and values obtained were comparable to those for EF. The similarity in the dissociation constants determined by SPR and by measurements on the cell surface suggests that the presence of the receptor does not play a large role in the interaction between PA(63) and EF/LF. PMID- 10828991 TI - CP1 domain in Escherichia coli leucyl-tRNA synthetase is crucial for its editing function. AB - The amino acid discrimination by aminoacyl-tRNA synthetase is achieved through two sifting steps; amino acids larger than the cognate substrate are rejected by a "coarse sieve", while the reaction products of amino acids smaller than the cognate substrate will go through a "fine sieve" and be hydrolyzed. This "double sieve" mechanism has been proposed for IleRS, a class I aminoacyl-tRNA synthetase. In this study, we created LeuRS-B, a mutant leucyl-tRNA synthetase from Escherichia coli with a duplication of the peptide fragment from Met328 to Pro368 (within its CP1 domain). This mutant has 50% of the leucylation activity of the wild-type enzyme and has the same ability to discriminate noncognate amino acids in the first step of the reaction. However, LeuRS-B can catalyze mischarging of tRNA(Leu) by methionine or isoleucine, suggesting that it is impaired in the ability to edit incorrect products. Wild-type leucyl-tRNA synthetase can edit the mischarged tRNA(Leu) made by LeuRS-B, while a separated CP1 domain cannot. These data suggest that the CP1 domain of leucyl-tRNA synthetase is crucial to the second editing sieve and that CP1 needs the structural context in leucyl-tRNA synthetase to fulfill its editing function. PMID- 10828990 TI - Secondary binding sites for triplex-forming oligonucleotides containing bulges, loops, and mismatches in the third strand. AB - We have used DNase I footprinting to examine the binding of five different 17-mer oligonucleotides to a 53-base oligopurine tract containing four pyrimidine interruptions. Although all the expected triplexes formed with high affinity (K(d) approximately 10-50 nM), one oligonucleotide produced a footprint at a second site with about 20-fold lower affinity. We have explored the nature of this secondary binding site and suggest that it arises when each end of the third strand forms a 7-mer triplex with adjacent regions on the duplex, generating a contiguous 14-base triplex with a bulge in the center of the third strand oligonucleotide. This unusual binding mode was examined by use of oligonucleotides that were designed with the potential to form different length third-strand loops of various base composition. We find that triplexes containing single-base bulges are generally more stable than those with dinucleotide loops, though triplexes can be formed with loops of up to nine thymines, generating complexes with submicromolar dissociation constants. These structures are much more stable than those formed by adding two separate 7-mer oligonucleotides, which do not generate DNase I footprints, though a stable complex is generated when the two halves are covalently joined by a hexa(ethylene glycol) linker. MPE produces less clear footprints, presumably because this cleavage agent binds to triplex DNA, but confirms that the oligonucleotides can bind in unexpected places. These results suggest that extra care needs to be taken when designing long triplex-forming oligonucleotides so as to avoid triplex formation at shorter secondary sites. PMID- 10828992 TI - Investigation of the DsbA mechanism through the synthesis and analysis of an irreversible enzyme-ligand complex. AB - Approaching the molecular mechanism of some enzymes is hindered by the difficulty of obtaining suitable protein-ligand complexes for structural characterization. DsbA, the major disulfide oxidase in the bacterial periplasm, is such an enzyme. Its structure has been well characterized in both its oxidized and its reduced states, but structural data about DsbA-peptide complexes are still missing. We report herein an original, straightforward, and versatile strategy for making a stable covalent complex with a cysteine-homoalanine thioether bond instead of the labile cystine disulfide bond which normally forms between the enzyme and polypeptides during the catalytic cycle of DsbA. We substituted a bromohomoalanine for the cysteine in a model 14-mer peptide derived from DsbB (PID-Br), the membrane partner of DsbA. When incubated in the presence of the enzyme, a selective nucleophilic substitution of the bromine by the thiolate of the DsbA Cys(30) occurred. The major advantage of this strategy is that it enables the direct use of the wild-type form of the enzyme, which is the most relevant to obtain unbiased information on the enzymatic mechanism. Numerous intermolecular NOEs between DsbA and PID could be observed by NMR, indicating the presence of preferential noncovalent interactions between the two partners. The thermodynamic properties of the DsbA-PID complex were measured by differential scanning calorimetry. In the complex, the values for both denaturation temperature and variation in enthalpy associated with thermal unfolding were between those of oxidized and reduced forms of DsbA. This progressive increase in stability along the DsbA catalytic pathway strongly supports the model of a thermodynamically driven mechanism. PMID- 10828993 TI - Inhibition of the human sodium/bile acid cotransporters by side-specific methanethiosulfonate sulfhydryl reagents: substrate-controlled accessibility of site of inactivation. AB - Mammalian sodium/bile acid cotransporters (SBATs) constitute a subgroup of the sodium cotransporter superfamily and function in the enterohepatic circulation of bile acids. They are glycoproteins with an exoplasmic N-terminus, seven or nine transmembrane segments, and a cytoplasmic C-terminus. They exhibit no significant homology with other members of the sodium cotransporter family and there is limited structure/function information available for the SBATs. Membrane impermeant methanethiosulfonates (MTS) inhibited bile acid transport by alkylation of cysteine 270 (apical SBAT)/266 (basolateral SBAT) that is fully conserved among the sodium/bile acid cotransporters. The accessibility of this residue to MTS reagent is regulated by the natural substrates, sodium and bile acid. In experiments with the apical SBAT, sodium alone increases the reactivity with the thiol reagents as compared to sodium-free medium. In contrast, bile acids protect the SBATs from inactivation, although only in the presence of sodium. The inhibition and protection data suggest that cysteine 270/266 lies in a sodium-sensitive region of the SBATs that is implicated in bile acid transport. PMID- 10828995 TI - Goodness-of-Fit Analysis of Radial Velocity Surveys. AB - Using eigenmode expansion of the Mark III and SFI surveys of cosmological radial velocities, a goodness-of-fit analysis is applied on a mode-by-mode basis. This differential analysis complements the Bayesian maximum likelihood analysis that finds the most probable model given the data. Analyzing the surveys with their corresponding most likely models from the CMB-like family of models, as well as with the currently popular LambdaCDM model, reveals a systematic inconsistency of the data with these "best" models. There is a systematic trend of the cumulative chi(2) to increase with the mode number (where the modes are sorted by decreasing order of the eigenvalues). This corresponds to a decrease of the chi(2) with the variance associated with a mode and hence with its effective scale. It follows that the differential analysis finds that on small (large) scales the global analysis of all the modes "puts" less (more) power than actually required by the data. This observed trend might indicate one of the following: (1) the theoretical model (i.e., power spectrum) or the error model (or both) have an excess of power on large scales, (2) velocity bias, or (3) the velocity data suffers from systematic errors that have not yet been corrected. PMID- 10828997 TI - A Comparison of Estimators for the Two-Point Correlation Function. AB - Nine of the most important estimators known for the two-point correlation function are compared using a predetermined, rigorous criterion. The indicators were extracted from over 500 subsamples of the Virgo Hubble volume simulation cluster catalog. The "real" correlation function was determined from the full survey in a 3000 h(-1) Mpc periodic cube. The estimators were ranked by the cumulative probability of returning a value within a certain tolerance of the real correlation function. This criterion takes into account bias and variance, and it is independent of the possibly non-Gaussian nature of the error statistics. As a result, for astrophysical applications, a clear recommendation has emerged: the Landy & Szalay estimator, in its original or grid version (Szapudi & Szalay), is preferred in comparison with the other indicators examined, with a performance almost indistinguishable from the Hamilton estimator. PMID- 10828994 TI - Cosmological Parameters from the Eigenmode Analysis of the Las Campanas Redshift Survey. AB - We present the first results of the Karhunen-Loeve eigenmodes applied to real data of the Las Campanas Redshift Survey to simultaneously measure the values of the redshift distortion parameter beta=Omega0.60&solm0;b, the linearly extrapolated normalization sigmaL8, and the cold dark matter shape parameter Gamma=Omega0h. The results of our numerical likelihood analysis indicate a low value of beta=0.30+/-0.39, a shape parameter Gamma=0.16+/-0.10, and a linearly extrapolated normalization sigmaL8=0.79+/-0.08, which are consistent with a low density universe (Omega0 less, similar0.5). PMID- 10828996 TI - Weak Lensing by Large-Scale Structure: A Dark Matter Halo Approach. AB - Weak gravitational lensing observations probe the spectrum and evolution of density fluctuations and the cosmological parameters that govern them, but they are currently limited to small fields and subject to selection biases. We show how the expected signal from large-scale structure arises from the contributions from and correlations between individual halos. We determine the convergence power spectrum as a function of the maximum halo mass and so provide the means to interpret results from surveys that lack high-mass halos either through selection criteria or small fields. Since shot noise from rare massive halos is mainly responsible for the sample variance below 10&arcmin;, our method should aid our ability to extract cosmological information from small fields. PMID- 10829000 TI - X-Ray Spectral Diagnostics of Gamma-Ray Burst Environments. AB - Recently, detection of discrete features in the X-ray afterglow spectra of GRB 970508 and GRB 970828 was reported. The most natural interpretation of these features is that they are redshifted Fe K emission complexes. The identification of the line emission mechanism has drastic implications for the inferred mass of radiating material and hence the nature of the burst site. X-ray spectroscopy provides a direct observational constraint on these properties of gamma-ray bursters. We briefly discuss how these constraints arise in the context of an application to the spectrum of GRB 970508. PMID- 10828999 TI - Collisional versus Collisionless Dark Matter. AB - We compare the structure and substructure of dark matter halos in model universes dominated by collisional, strongly self-interacting dark matter (SIDM) and collisionless, weakly interacting dark matter (CDM). While SIDM virialized halos are more nearly spherical than CDM halos, they can be rotationally flattened by as much as 20% in their inner regions. Substructure halos suffer ram-pressure truncation and drag, which are more rapid and severe than their gravitational counterparts tidal stripping and dynamical friction. Lensing constraints on the size of galactic halos in clusters are a factor of 2 smaller than predicted by gravitational stripping, and the recent detection of tidal streams of stars escaping from the satellite galaxy Carina suggests that its tidal radius is close to its optical radius of a few hundred parsecs-an order of magnitude smaller than predicted by CDM models but consistent with SIDM models. The orbits of SIDM satellites suffer significant velocity bias, sigmaSIDM&solm0;sigmaCDM=0.85, and are more circular than CDM satellites, betaSIDM approximately 0.5, in agreement with the inferred orbits of the Galaxy's satellites. In the limit of a short mean free path, SIDM halos have singular isothermal density profiles; thus, in its simplest incarnation SIDM, is inconsistent with galactic rotation curves. PMID- 10829001 TI - Spectral Lags of Gamma-Ray Bursts From Ginga and BATSE. AB - The analysis of spectral lag between energy bands, which combines temporal and spectral analyses, can add strict constraints to gamma-ray burst (GRB) models. In previous studies, the lag analysis focused on the lags between channel 1 (25-57 keV) and channel 3 (115-320 keV) from the Burst and Transient Source Experiment (BATSE). In this Letter, we analyzed the cross-correlation average lags (including approximate uncertainties) between energy bands for two GRB samples: 19 events detected by Ginga and 109 events detected by BATSE. We paid special attention to the BATSE GRBs with known redshifts because there has been a reported connection between lag and luminosity. This extends our knowledge of spectral lags to lower energy ( approximately 2 keV). We found that lags between energy bands are small. The lag between the peak of approximately 50 keV photons and that of approximately 200 keV photons is approximately 0.08 s. The upper limit in the lag between approximately 9 and approximately 90 keV photons is approximately 0.5 s. Thus, there are not large shifts at low energy. We found that about 20% of GRBs have detectable lags between energy bands in the Ginga and BATSE samples. From the internal shock model, we found that there are three sources of time structure in GRB pulses: cooling, hydrodynamics, and angular effects. We argue that cooling is much too fast to account for our observed lags and that angular effects are independent of energy. Thus, only hydrodynamics can produce these lags. Perhaps the radiation process varies as the reverse shock moves through the shell. PMID- 10828998 TI - Discovery of Narrow X-Ray Absorption Lines from NGC 3783 with the Chandra High Energy Transmission Grating Spectrometer. AB - We present the first grating-resolution X-ray spectra of the Seyfert 1 galaxy NGC 3783, obtained with the High Energy Transmission Grating Spectrometer on the Chandra X-Ray Observatory. These spectra reveal many narrow absorption lines from the H-like and He-like ions of O, Ne, Mg, Si, S, and Ar as well as Fe xvii-Fe xxi L-shell lines. We have also identified several weak emission lines, mainly from O and Ne. The absorption lines are blueshifted by a mean velocity of approximately 440+/-200 km s-1 and are not resolved, indicating a velocity dispersion within the absorbing gas of a few hundred kilometers per second or less. We measure the lines' equivalent widths and compare them with the predictions of photoionization models. The best-fitting model has a microturbulence velocity of 150 km s-1 and a hydrogen column density of 1.3x1022 cm-2. The measured blueshifts and inferred velocity dispersions of the X-ray absorption lines are consistent with those of the strongest UV absorption lines observed in this object. However, simple models that propose to strictly unify the X-ray and UV absorbers have difficulty explaining simultaneously the X-ray and UV absorption-line strengths. PMID- 10829003 TI - Smoothed Particle Hydrodynamics Simulations of Apsidal and Nodal Superhumps. AB - In recent years, a handful of systems have been observed to show "negative" (nodal) superhumps, with periods slightly shorter than the orbital period. It has been suggested that these modes are a consequence of the slow retrograde precession of the line of nodes in a disk tilted with respect to the orbital plane. Our simulations confirm and refine this model: they suggest a roughly axisymmetric, retrogradely precessing, tilted disk that is driven at a period slightly less than half the orbital period as the tidal field of the orbiting secondary encounters, in turn, the two halves of the disk above and below the midplane. Each of these passings leads to viscous dissipation on one face of an optically thick disk-observers on opposite sides of the disk would each observe one brightening per orbit, but 180 degrees out of phase with each other. PMID- 10829002 TI - Impulsive and Varying Injection in Gamma-Ray Burst Afterglows. AB - The standard model of gamma-ray burst afterglows is based on synchrotron radiation from a blast wave produced when the relativistic ejecta encounters the surrounding medium. We reanalyze the refreshed shock scenario, in which slower material catches up with the decelerating ejecta and reenergizes it. This energization can be done either continuously or in discrete episodes. We show that such a scenario has two important implications. First, there is an additional component coming from the reverse shock that goes into the energizing ejecta. This persists for as long as the reenergization itself, which could extend for up to days or longer. We find that during this time the overall spectral peak is found at the characteristic frequency of the reverse shock. Second, if the injection is continuous, the dynamics will be different from that in constant energy evolution and will cause a slower decline of the observed fluxes. A simple test of the continuously refreshed scenario is that it predicts a spectral maximum in the far-infrared or millimeter range after a few days. PMID- 10829004 TI - Diffusion in the Atmospheres of Blue Horizontal-Branch Stars. AB - We investigate the effects of diffusion in the atmospheres of hot horizontal branch stars using a model atmosphere code including diffusion self-consistently. Equilibrium stratifications (i.e., for which the diffusion velocity equals zero in each layer) are computed for models of effective temperatures between 10,000 and 25,000 K. The stratified models provide much better agreement with many observational features [jump in the (u, u-y) color-magnitude diagram, gaps, lower spectroscopic gravities] in comparison with classical horizontal-branch models. The observed abundance anomalies are also consistent with the amounts that can be supported in the atmospheres. PMID- 10829005 TI - Disk Accretion in the 10 Myr Old T Tauri Stars TW Hydrae and Hen 3-600A. AB - We have found that two members of the TW Hydrae association, TW Hydrae and Hen 3 600A, are still actively accreting, based on the ballistic infall signature of their broad Halpha emission profiles. We present the first quantitative analysis of accretion in these objects and conclude that the same accretion mechanisms which operate in the well-studied 1 Myr old T Tauri stars can and do occur in older (10 Myr) stars. We derive the first estimates of the disk mass accretion rate in TW Hya and Hen 3-600A, which are 1-2 orders of magnitude lower than the average rates in 1 Myr old objects. The decrease in accretion rates over 10 Myr, as well as the low fraction of TW Hya association objects still accreting, points to significant disk evolution, possibly linked to planet formation. Given the multiplicity of the Hen 3-600 system and the large UV excess of TW Hya, our results show that accretion disks can be surprisingly long lived in spite of the presence of companions and significant UV ionizing flux. PMID- 10829006 TI - High Magnetic Field Pulsars and Magnetars: A Unified Picture. AB - We propose a unified picture of high magnetic field radio pulsars and magnetars by arguing that they are all rotating high-field neutron stars but that their magnetic axes have different orientations with respect to their rotation axes. In strong magnetic fields where photon splitting suppresses pair creation near the surface, the high-field pulsars can have active inner accelerators while the anomalous X-ray pulsars cannot. This can account for the very different observed emission characteristics of the anomalous X-ray pulsar 1E 2259+586 and the high field radio pulsar PSR J1814-1744. A predicted consequence of this picture is that radio pulsars having surface magnetic fields greater than about 2x1014 G should not exist. PMID- 10829007 TI - Timing Noise in SGR 1806-20. AB - We have phase-connected a sequence of Rossi X-Ray Timing Explorer Proportional Counter Array observations of SGR 1806-20 covering 178 days. We find that a simple secular spin-down model does not adequately fit the data. The period derivative varies gradually during the observations between 8.1x10-11 and 11.7x10 11 s s(-1) (at its highest, approximately 40% larger than the long-term trend), while the average burst rate as seen with the Burst and Transient Source Experiment drops throughout the time interval. The phase residuals give no compelling evidence for periodicity, but more closely resemble timing noise as seen in radio pulsars. The magnitude of the timing noise, however, is large relative to the noise level typically found in radio pulsars (Delta8=4.8; frequency derivative average power approximately 7x10-20 cycles(2) s(-3)). Combining these results with the noise levels measured for some anomalous X-ray pulsars, we find that all magnetar candidates have Delta(8) values larger than those expected from a simple extrapolation of the correlation found in radio pulsars. We find that the timing noise in SGR 1806-20 is greater than or equal to the levels found in some accreting systems (e.g., Vela X-1, 4U 1538-52, and 4U 1626-67), but the spin-down of SGR 1806-20 has thus far maintained coherence over 6 yr. Alternatively, an orbital model with a period Porb=733 days provides a statistically acceptable fit to the data. If the phase residuals are created by Doppler shifts from a gravitationally bound companion, then the allowed parameter space for the mass function (small) and orbital separation (large) rule out the possibility of accretion from the companion sufficient to power the persistent emission from the SGR. PMID- 10829008 TI - The Effect of Hydrostatic Weighting on the Vertical Temperature Structure of the Solar Corona. AB - We investigate the effect of hydrostatic scale heights lambda(T) in coronal loops on the determination of the vertical temperature structure T&parl0;h&parr0; of the solar corona. Every method that determines an average temperature at a particular line of sight from optically thin emission (e.g., in EUV or soft X-ray wavelengths) of a mutlitemperature plasma is subject to the emission measure weighted contributions dEM&parl0;T&parr0;&solm0;dT from different temperatures. Because most of the coronal structures (along open or closed field lines) are close to hydrostatic equilibrium, the hydrostatic temperature scale height introduces a height-dependent weighting function that causes a systematic bias in the determination of the temperature structure T&parl0;h&parr0; as function of altitude h. The net effect is that the averaged temperature seems to increase with altitude, dT&parl0;h&parr0;&solm0;dh>0, even if every coronal loop (of a multitemperature ensemble) is isothermal in itself. We simulate this effect with differential emission measure distributions observed by SERTS for an instrument with a broadband temperature filter such as Yohkoh/Soft X-Ray Telescope and find that the apparent temperature increase due to hydrostatic weighting is of order DeltaT approximately T0h&solm0;r middle dot in circle. We suggest that this effect largely explains the systematic temperature increase in the upper corona reported in recent studies (e.g., by Sturrock et al., Wheatland et al., or Priest et al.), rather than being an intrinsic signature of a coronal heating mechanism. PMID- 10829009 TI - Small Scale Structure in the Solar Transition Region. AB - We present analysis of a high spatial resolution ( approximately 1&farcs;5) O vi lambda1032 spectroheliogram taken with the Solar Ultraviolet Measurements of Emitted Radiation (SUMER) spectrometer on the Solar and Heliospheric Observatory. This image shows numerous narrow, looplike structures. The widths of the narrowest resolved features are at the spatial resolution of the SUMER spectrometer. In contrast to earlier Skylab observations, however, comparisons with a Michelson Doppler Imager magnetogram reveals that the majority of these looplike structures do not connect network magnetic fields. Instead, they extend from the supergranulation network into the cell-center regions where no magnetic fields are detected in these data. We conjecture that these fine structures are related to spicules and are highly dynamic. PMID- 10829010 TI - Excitation of Oscillations in the Magnetic Network on the Sun. AB - We examine the excitation of oscillations in the magnetic network of the Sun through the footpoint motion of photospheric magnetic flux tubes located in intergranular lanes. The motion is derived from a time series of high-resolution G-band and continuum filtergrams using an object-tracking technique. We model the response of the flux tube to the footpoint motion in terms of the Klein-Gordon equation, which is solved analytically as an initial value problem for transverse (kink) waves. We compute the wave energy flux in upward-propagating transverse waves. In general we find that the injection of energy into the chromosphere occurs in short-duration pulses, which would lead to a time variability in chromospheric emission that is incompatible with observations. Therefore, we consider the effects of turbulent convective flows on flux tubes in intergranular lanes. The turbulent flows are simulated by adding high-frequency motions (periods 5-50 s) with an amplitude of 1 km s(-1). The latter are simulated by adding random velocity fluctuations to the observationally determined velocities. In this case, we find that the energy flux is much less intermittent and can in principle carry adequate energy for chromospheric heating. PMID- 10829011 TI - Translational incorporation of S-nitrosohomocysteine into protein. AB - The non-protein amino acid homocysteine (Hcy), owing to its structural similarity to the protein amino acids methionine, isoleucine, and leucine, enters first steps of protein synthesis and is activated by methionyl-, isoleucyl-, and leucyl tRNA synthetases in vivo. However, translational incorporation of Hcy into protein is prevented by editing mechanisms of these synthetases, which convert misactivated Hcy into thiolactone. The lack of efficient interactions of the side chain of Hcy with the specificity subsite of the synthetic/editing active site is a prerequisite for editing of Hcy. Thus, if the side chain thiol of Hcy were reversibly modified with a small molecule that would enhance its binding to the specificity subsite and prevent editing, such modified Hcy is predicted to be transferred to tRNA and incorporated translationally into protein. Here I show that S-nitroso-Hcy is in fact transferred to tRNA by methionyl-tRNA synthetase and incorporated into protein by the bacterium Escherichia coli. S-Nitroso-Hcy tRNA also supports translation of mRNAs in a rabbit reticulocyte system. Removal of the nitroso group yields Hcy-tRNA and protein containing Hcy in peptide bonds. S-Nitrosylation-mediated translational incorporation of Hcy into protein may occur under natural conditions in cells and contribute to Hcy-induced pathogenesis in atherosclerosis. PMID- 10829012 TI - Binding of GDNF and neurturin to human GDNF family receptor alpha 1 and 2. Influence of cRET and cooperative interactions. AB - The members of the glial cell line-derived neurotrophic factor (GDNF) family signal via binding to the glycosyl phosphatidylinositol-anchored membrane proteins, the GDNF family receptors alpha (GFRalpha), and activation of cRET. We performed a detailed analysis of the binding of GDNF and neurturin to their receptors and investigated the influence of cRET on the binding affinities. We show that the rate of dissociation of (125)I-GDNF from GFRalpha1 is increased in the presence of 50 nm GDNF, an effect that can be explained by the occurrence of negative cooperativity. Scatchard plots of the ligand concentration binding isotherms reveal a pronounced downward curvature at low (125)I-GDNF concentrations suggesting the presence of positive cooperativity. This effect is observed in the range of GDNF concentrations responsible for biological activity (1-20 pm) and may have an important role in cRET-independent signaling. A high affinity site with a K(D) of 11 pm for (125)I-GDNF is detected only when GFRalpha1 is co-expressed with cRET at a DNA ratio of 1:3. These results suggest an interaction of GFRalpha1 and cRET in the absence of GDNF and demonstrate that the high affinity binding can be measured only when cRET is present. PMID- 10829014 TI - Structural and functional role of the disulfide bridges in the hydrophobin SC3. AB - Hydrophobins function in fungal development by self-assembly at hydrophobic hydrophilic interfaces such as the interface between the fungal cell wall and the air or a hydrophobic solid. These proteins contain eight conserved cysteine residues that form four disulfide bonds. To study the effect of the disulfide bridges on the self-assembly, the disulfides of the SC3 hydrophobin were reduced with 1,4-dithiothreitol. The free thiols were then blocked with either iodoacetic acid (IAA) or iodoacetamide (IAM), introducing eight or zero negative charges, respectively. Circular dichroism and infrared spectroscopy showed that after opening of the disulfide bridges SC3 is initially unfolded. IAA-SC3 did not self assemble at the air-water interface upon shaking an aqueous solution. Remarkably, after drying down IAA-SC3 or after exposing it to Teflon, it refolded into a structure similar to that observed for native SC3 at these interfaces. Iodoacetamide-SC3 on the other hand, which does not contain extra charges, spontaneously refolded in water in the amyloid-like beta-sheet conformation, characteristic for SC3 assembled at the water-air interface. From this we conclude that the disulfide bridges of SC3 are not directly involved in self assembly but keep hydrophobin monomers soluble in the fungal cell or its aqueous environment, preventing premature self-assembly. PMID- 10829013 TI - Activation of apolipoprotein AI gene expression by protein kinase A and kinase C through transcription factor, Sp1. AB - Our previous finding that insulin induces apolipoprotein AI (apoAI) transcription points to the participation of intracellular signaling. This finding prompted us to ask whether two classical G-protein-coupled signaling pathways requiring activated protein kinase A (PKA) or kinase C (PKC) may also regulate apoAI. Therefore, human hepatoma, Hep G2 cells stably transfected with pAI.474-CAT, a reporter construct spanning -474 to -7 of apoAI DNA fused to chloramphenicol acetyltransferase (CAT) were treated with 10 microm forskolin (FSK) or 50 nm phorbol dibutyrate (PDBu) to activate PKA and PKC, respectively. Results showed that the apoAI promoter activity increased 4-5-fold following 24 h of treatment with either FSK or PDBu. Induction by either agent was blocked with actinomycin D but not the protein synthesis inhibitor, cycloheximide. The PKA inhibitor, PKI 14 22 amide, abrogated induction by FSK, 100 microm 8-bromo-cAMP, or 100 ng/ml cholera toxin, but it had no effect on activation via PKC. Similarly, PDBu induction was attenuated by 2 microm of the PKC inhibitor, GF109203X, but it did not affect FSK activity. Next we used deletional constructs to show that the actions of FSK and PDBu required the insulin-responsive core element (IRCE). This motif matched the consensus binding site for the transcription factor, Sp1. The binding of Sp1 to the IRCE was confirmed by gel-retardation and supershift analysis. Site-directed mutagenesis of the IRCE eliminated Sp1 action and induction by FSK or PDBu. Whereas overexpression of Sp1 enhanced basal and FSK or PDBu induced promoter activity, transfection of an antisense oligomer against Sp1 mRNA attenuated both parameters. In summary, activation of PKA or PKC increases apoAI promoter activity. The activity of both signaling pathways is mediated by the IRCE, a motif that binds the transcription factor, Sp1. PMID- 10829015 TI - A novel human tocopherol-associated protein: cloning, in vitro expression, and characterization. AB - Vitamin E (alpha-tocopherol) is an essential dietary nutrient for humans and animals. The mechanisms involved in cellular regulation as well as in the preferential cellular and tissue accumulation of alpha-tocopherol are not yet well established. We previously reported (Stocker, A., Zimmer, S., Spycher, S. E., and Azzi, A. (1999) IUBMB Life 48, 49-55) the identification of a novel 46 kDa tocopherol-associated protein (TAP) in the cytosol of bovine liver. Here, we describe the identification, the molecular cloning into Escherichia coli, and the in vitro expression of the human homologue of bovine TAP, hTAP. This protein appears to belong to a family of hydrophobic ligand binding proteins, which have the CRAL (cis-retinal binding motif) sequence in common. By using a biotinylated alpha-tocopherol derivative and the IASys resonant mirror biosensor, the purified recombinant protein was shown to bind tocopherol at a specific binding site with K(d) 4.6 x 10(-7) m. Northern analyses showed that hTAP mRNA has a size of approximately 2800 base pairs and is ubiquitously expressed. The highest amounts of hTAP message are found in liver, brain, and prostate. In conclusion, hTAP has sequence homology to proteins containing the CRAL_TRIO structural motif. TAP binds to alpha-tocopherol and biotinylated tocopherol, suggesting the existence of a hydrophobic pocket, possibly analogous to that of SEC14. PMID- 10829016 TI - Escherichia coli NifS-like proteins provide selenium in the pathway for the biosynthesis of selenophosphate. AB - Selenophosphate synthetase (SPS), the selD gene product from Escherichia coli, catalyzes the biosynthesis of monoselenophosphate, AMP, and orthophosphate in a 1:1:1 ratio from selenide and ATP. Kinetic characterization revealed the K(m) value for selenide approached levels that are toxic to the cell. Our previous demonstration that a Se(0)-generating system consisting of l-selenocysteine and the Azotobacter vinelandii NifS protein can replace selenide for selenophosphate biosynthesis in vitro suggested a mechanism whereby cells can overcome selenide toxicity. Recently, three E. coli NifS-like proteins, CsdB, CSD, and IscS, have been overexpressed and characterized. All three enzymes act on selenocysteine and cysteine to produce Se(0) and S(0), respectively. In the present study, we demonstrate the ability of each E. coli NifS-like protein to function as a selenium delivery protein for the in vitro biosynthesis of selenophosphate by E. coli wild-type SPS. Significantly, the SPS (C17S) mutant, which is inactive in the standard in vitro assay with selenide as substrate, was found to exhibit detectable activity in the presence of CsdB, CSD, or IscS and l-selenocysteine. Taken together the ability of the NifS-like proteins to generate a selenium substrate for SPS and the activation of the SPS (C17S) mutant suggest a selenium delivery function for the proteins in vivo. PMID- 10829017 TI - Quantitative analysis of TIP47-receptor cytoplasmic domain interactions: implications for endosome-to-trans Golgi network trafficking. AB - TIP47 (tail-interacting protein of 47 kDa) binds to the cytoplasmic domains of the cation-independent and cation-dependent mannose 6-phosphate receptors and is required for their transport from late endosomes to the trans Golgi network in vitro and in vivo. We report here a quantitative analysis of the interaction of recombinant TIP47 with mannose 6-phosphate receptor cytoplasmic domains. Recombinant TIP47 binds more tightly to the cation-independent mannose 6 phosphate receptor (K(D) = 1 microm) than to the cation-dependent mannose 6 phosphate receptor (K(D) = 3 microm). In addition, TIP47 fails to interact with the cytoplasmic domains of the hormone-processing enzymes, furin, phosphorylated furin, and metallocarboxypeptidase D, as well as the cytoplasmic domain of TGN38, proteins that are also transported from endosomes to the trans Golgi network. Although these proteins failed to bind TIP47, furin and TGN38 were readily recognized by the clathrin adaptor, AP-2. These data suggest that TIP47 recognizes a very select set of cargo molecules. Moreover, our data suggest unexpectedly that furin, TGN38, and carboxypeptidase D may use a distinct vesicular carrier and perhaps a distinct route for transport between endosomes and the trans Golgi network. PMID- 10829018 TI - The receptor for advanced glycation end products is induced by the glycation products themselves and tumor necrosis factor-alpha through nuclear factor-kappa B, and by 17beta-estradiol through Sp-1 in human vascular endothelial cells. AB - The binding of advanced glycation end products (AGE) to the receptor for AGE (RAGE) is known to deteriorate various cell functions and is implicated in the pathogenesis of diabetic vascular complications. Here we show that AGE, tumor necrosis factor-alpha (TNF-alpha), and 17beta-estradiol (E(2)) up-regulated RAGE mRNA and protein levels in human microvascular endothelial cells and ECV304 cells, with the mRNA stability being essentially invariant. Transient transfection experiments with human RAGE promoter-luciferase chimeras revealed that the region from nucleotide number -751 to -629 and the region from -239 to 89 in the RAGE 5'-flanking sequence exhibited the AGE/TNF-alpha and E(2) responsiveness, respectively. Site-directed mutation of an nuclear factor-kappaB (NF-kappaB) site at -671 or of Sp-1 sites at -189 and -172 residing in those regions resulted in an abrogation of the AGE/TNF-alpha- or E(2)-mediated transcriptional activation. Electrophoretic mobility shift assays revealed that ECV304 cell nuclear extracts contained factors which retarded the NF-kappaB and Sp-1 elements, and that the DNA-protein complexes were supershifted by anti p65/p50 NF-kappaB and anti-Sp-1/estrogen receptor alpha antibodies, respectively. These results suggest that AGE, TNF-alpha, and E(2) can activate the RAGE gene through NF-kappaB and Sp-1, causing enhanced AGE-RAGE interactions, which would lead to an exacerbation of diabetic microvasculopathy. PMID- 10829019 TI - Cloning, expression, and functional characterization of the substrate binding subunit of rat type II iodothyronine 5'-deiodinase. AB - Type II iodothyronine 5'-deiodinase catalyzes the bioactivation of thyroid hormone in the brain. In astrocytes, this approximately 200-kDa, membrane-bound enzyme is composed of at least one p29 subunit, an approximately 60-kDa, cAMP induced activation protein, and one or more unidentified catalytic subunit(s). Recently, an artificial type II-like selenodeiodinase was engineered by fusing two independent cDNAs together; however, no native type II selenodeiodinase polypeptide is translated in the brain or brown adipose tissue of rats. These data suggest that the native type II 5'-deiodinase in rat brain is unrelated to this artificial selenoprotein. In this report, we describe the cloning of the 29 kDa subunit (p29) of type II 5'-deiodinase from a lambdazapII cDNA library prepared from cAMP-induced astrocytes. The 3.3-kilobase (kb) cDNA encodes an approximately 30-kDa, 277-amino acid long, hydrophobic protein lacking selenocysteine. Northern blot analysis showed that a 3.5-kb p29 mRNA was present in tissues showing type II 5'-deiodinase activity such as brain and cAMP stimulated astrocytes. Domain-specific, anti-p29 antibodies specifically immunoprecipitated enzyme activity. Overexpression of exogenous p29 or a green fluorescence protein (GFP)-tagged p29 fusion protein led to a >100-fold increase in deiodinating activity in cAMP-stimulated astrocytes, and the increased activity was specifically immunoprecipitated by anti-GFP antibodies. Steady-state reaction kinetics of the enzyme in GFP-tagged p29-expressing astrocytes are identical to those of the native enzyme in brain. Direct injection of replication deficient Ad5-p29(GFP) virus particles into the cerebral cortex of neonatal rats leads to a approximately 2-fold increase in brain type II 5'-deiodinating activity. These data show 1) that the 3.3-kb p29 cDNA encodes an essential subunit of rat type II iodothyronine 5'-deiodinase and 2) identify the first non selenocysteine containing subunit of the deiodinase family of enzymes. PMID- 10829020 TI - Dimerization choices control the ability of axin and dishevelled to activate c Jun N-terminal kinase/stress-activated protein kinase. AB - Axin and Dishevelled are two downstream components of the Wnt signaling pathway. Dishevelled is a positive regulator and is placed genetically between Frizzled and glycogen synthase kinase-3beta, whereas Axin is a negative regulator that acts downstream of glycogen synthase kinase-3beta. It is intriguing that they each can activate the c-Jun N-terminal kinase/stress-activated protein kinase (JNK/SAPK) when expressed in the cell. We set out to address if Axin and Dishevelled are functionally cooperative, antagonistic, or entirely independent, in terms of the JNK activation event. We found that in contrast to Axin, Dvl2 activation of JNK does not require MEKK1, and complex formation between Dvl2 and Axin is independent of Axin-MEKK1 binding. Furthermore, Dvl2-DIX and Dvl2 DeltaDEP proteins deficient for JNK activation can attenuate Axin-activated JNK activity by disrupting Axin dimerization. However, Axin-DeltaMID, Axin-DeltaC, and Axin-CT proteins deficient for JNK activation cannot interfere with Dvl2 activated JNK activity. These results indicate that unlike the strict requirement of homodimerization for Axin function, Dvl2 can activate JNK either as a monomer or homodimer/heterodimer. We suggest that there may be a switch mechanism based on dimerization combinations, that commands cells to activate Wnt signaling or JNK activation, and to turn on specific activators of JNK in response to various environmental cues. PMID- 10829021 TI - GTPases and phosphatidylinositol 3-kinase are critical for insulin-like growth factor-I-mediated Schwann cell motility. AB - Previously, we reported insulin-like growth factor-I (IGF-I) promotes motility and focal adhesion kinase (FAK) activation in neuronal cells. In the current study, we examined the role of IGF-I in Schwann cell (SC) motility. IGF-I increases SC process extension and motility. In parallel, IGF-I activates IGF-I receptor, insulin receptor substrate-1 (IRS-1), phosphatidylinositol 3 (PI-3) kinase, and FAK. LY294002, a PI-3 kinase inhibitor, blocks IGF-I-induced motility and FAK phosphorylation. The Rho family of GTPases is important in the regulation of the cytoskeleton. Overexpression of constitutively active Leu-61 Cdc42 and Val 12 Rac1 enhances SC motility which is unaffected by LY294002. In parallel, stable transfection of SC with dominant negative Asn-17 Rac1 blocks IGF-I-mediated SC motility and FAK phosphorylation, implying Rac is an upstream regulator of FAK. Collectively our results suggest that IGF-I regulates SC motility by reorganization of the actin cytoskeleton via the downstream activation of a PI-3 kinase, small GTPase, and FAK pathway. PMID- 10829022 TI - Theoretical calculation of pKa reveals an important role of Arg205 in the activity and stability of Streptomyces sp. N174 chitosanase. AB - Based on the crystal structure of chitosanase from Streptomyces sp. N174, we have calculated theoretical pK(a) values of the ionizable groups of this protein using a combination of the boundary element method and continuum electrostatics. The pK(a) value obtained for Arg(205), which is located in the catalytic cleft, was abnormally high (>20.0), indicating that the guanidyl group may interact strongly with nearby charges. Chitosanases possessing mutations in this position (R205A, R205H, and R205Y), produced by Streptomyces lividans expression system, were found to have less than 0.3% of the activity of the wild type enzyme and to possess thermal stabilities 4-5 kcal/mol lower than that of the wild type protein. In the crystal structure, the Arg(205) side chain is in close proximity to the Asp(145) side chain (theoretical pK(a), -1.6), which is in turn close to the Arg(190) side chain (theoretical pK(a), 17.7). These theoretical pK(a) values are abnormal, suggesting that both of these residues may participate in the Arg(205) interaction network. Activity and stability experiments using Asp(145)- and Arg(190)-mutated chitosanases (D145A and R190A) provide experimental data supporting the hypothesis derived from the theoretical pK(a) data and prompt the conclusion that Arg(205) forms a strong interaction network with Asp(145) and Arg(190) that stabilizes the catalytic cleft. PMID- 10829023 TI - Identification of residues within two regions involved in self-association of viral histone-like protein p6 from phage theta29. AB - Protein p6 of Bacillus subtilis phage theta29 is involved in the initiation of viral DNA replication and transcription by forming a multimeric nucleoprotein complex with the phage DNA. Based on this, together with its abundance and its capacity to bind to the whole viral genome, it has been proposed to be a viral histone-like protein. Protein p6 is in a monomer-dimer-oligomer equilibrium association. We have identified protein p6 mutants deficient in self-association by testing random mutants obtained by degenerated polymerase chain reaction in an in vivo assay for dimer formation. The mutations were mainly clustered in two regions located at the N terminus, and the central part of the protein. Site directed single mutants, corresponding to those found in vivo, have been constructed and purified. Mutant p6A44V, located at the central part of the protein, showed an impaired dimer formation ability, and a reduced capacity to bind DNA and to activate the initiation of O29 DNA replication. Mutant p6I8T has at least 10-fold reduced self-association capacity, does not bind DNA nor activate O29 DNA initiation of replication. C-terminal deletion mutants showed an enhanced dimer formation capacity. The highly acidic tail, removed in these mutants, is proposed to modulate the protein p6 self-association. PMID- 10829024 TI - Identification of a GA-rich sequence as a protein-binding site in the 3' untranslated region of chicken elastin mRNA with a potential role in the developmental regulation of elastin mRNA stability. AB - Synthesis of aortic elastin peaks in the perinatal period and then is strongly down-regulated with postnatal development and growth. Decreased stability of elastin mRNA contributes to this developmental decrease in chick aortic elastin production. We have previously shown that destabilization of elastin mRNA is correlated with decreased binding of cytosolic protein(s) to a large, GC-rich region of secondary structure in the 3'-untranslated region (3'-UTR) of elastin mRNA. In this study, using gel migration shift assays, deletion constructs, and antisense competition assays, we identify a major protein-binding site in the 3' UTR of elastin as a GA-rich sequence (UGGGGGGAGGGAGGGAGGGA), which we have designated the G3A motif. This motif is present in the 3'-UTR of elastin from several species. Binding proteins are present in both nuclear and cytoplasmic extracts, and their abundance is associated with tissues producing elastin and correlated with circumstances in which elastin mRNA is stable. These results suggest that the conserved GA-rich sequence of the elastin 3'-UTR is an important element in the regulation of stability of the elastin mRNA. PMID- 10829025 TI - Fibrillarin genes encode both a conserved nucleolar protein and a novel small nucleolar RNA involved in ribosomal RNA methylation in Arabidopsis thaliana. AB - Fibrillarin is a key nucleolar protein in eukaryotes which associates with box C/D small nucleolar RNAs (snoRNAs) directing 2'-O-ribose methylation of the rRNA. In this study we describe two genes in Arabidopsis thaliana, AtFib1 and AtFib2, encoding nearly identical proteins conserved with eukaryotic fibrillarins. We demonstrate that AtFib1 and AtFib2 proteins are functional homologs of the yeast Nop1p (fibrillarin) and can rescue a yeast NOP1-null mutant strain. Surprisingly, for the first time in plants, we identified two isoforms of a novel box C/D snoRNA, U60.1f and U60.2f, nested in the fifth intron of AtFib1 and AtFib2. Interestingly after gene duplication the host intronic sequences completely diverged, but the snoRNA was conserved, even in other crucifer fibrillarin genes. We show that the U60f snoRNAs accumulate in seedlings and that their targeted residue on the 25 S rRNA is methylated. Our data reveal that the three modes of expression of snoRNAs, single, polycistronic, and intronic, exist in plants and suggest that the mechanisms directing rRNA methylation, dependent on fibrillarin and box C/D snoRNAs, are evolutionarily conserved in plants. PMID- 10829026 TI - Mapping the functional domains of nucleolar protein B23. AB - Protein B23 is a multifunctional nucleolar protein whose cellular location and characteristics strongly suggest that it is a ribosome assembly factor. The protein has nucleic acid binding, ribonuclease, and molecular chaperone activities. To determine the contributions of unique polypeptide segments enriched in certain classes of amino acid residues to the respective activities, several constructs that produced N- and C-terminal deletion mutant proteins were prepared. The C-terminal quarter of the protein was shown to be necessary and sufficient for nucleic acid binding. Basic and aromatic segments at the N- and C terminal ends, respectively, of the nucleic acid binding region were required for activity. The molecular chaperone activity was contained in the N-terminal half of the molecule, with important contributions from both nonpolar and acidic regions. The chaperone activity also correlated with the ability of the protein to form oligomers. The central portion of the molecule was required for ribonuclease activity and possibly contains the catalytic site; this region overlapped with the chaperone-containing segment of the molecule. The C-terminal, nucleic acid-binding region enhanced the ribonuclease activity but was not essential for it. These data suggest that the three activities reside in mainly separate but partially overlapping segments of the polypeptide chain. PMID- 10829027 TI - In vitro translation of the upstream open reading frame in the mammalian mRNA encoding S-adenosylmethionine decarboxylase. AB - The upstream open reading frame (uORF) in the mRNA encoding S-adenosylmethionine decarboxylase is a polyamine-responsive element that suppresses translation of the associated downstream cistron in vivo. In this paper, we provide the first direct evidence of peptide synthesis from the S-adenosylmethionine decarboxylase uORF using an in vitro translation system. We examine both the influence of cation concentration on peptide synthesis and the effect of altering the uORF sequence on peptide synthesis. Synthesis of wild type and altered peptides was similar at all concentrations of magnesium tested. In contrast, synthesis of the wild type peptide was more sensitive than that of altered peptides to elevated concentrations of the naturally occurring polyamines, spermidine and spermine, as well as several polyamine analogs. The sensitivity of in vitro synthesis to spermidine was influenced by both the amino acid sequence and the length of the peptide product of the uORF. Findings from the present study correlate with the effects of the uORF and polyamines on translation of a downstream cistron in vivo and support the hypothesis that polyamines and the structure of the nascent peptide create a rate-limiting step in uORF translation, perhaps through a ribosome stalling mechanism. PMID- 10829028 TI - Activation of serum response factor in the depolarization induction of Egr-1 transcription in pancreatic islet beta-cells. AB - The results of the current studies define the major elements whereby glucose metabolism in islet beta-cells leads to transcriptional activation of an early response gene in insulinoma cell lines and in rat islets. Glucose stimulation (2 20 mm) resulted in a 4-fold increase in Egr-1 mRNA at 30 min, as did the depolarizing agents KCl and tolbutamide. This response was inhibited by diazoxide and EGTA, indicating that beta-cell depolarization and Ca(2+) influx, respectively, are essential. Pharmacological inhibition of the Egr-1 induction by H89 (48%) and calmidazolium (35%), but not by mitogen-activated protein kinase/extracellular signal-regulated kinase kinase 1 and 2 or phosphatidylinositol 3-kinase inhibitors, implied that protein kinase A and Ca(2+)/calmodulin pathways are involved. Deletion mapping of the Egr-1 promoter revealed that the proximal -198 base pairs containing two serum response elements (SREs) and one cAMP-response element retained the depolarization response. Depolarization resulted in phosphorylation of cAMP-response element-binding protein, yet partial inhibition by a dominant negative cAMP-response element binding protein, along with a robust response of a cAMP-response element-mutated Egr-1 promoter suggested the presence of a second Ca(2+)-responsive element. Depolarization activation of 5XSRE-LUC and serum response factor (SRF)-GAL4 constructs, along with activation of SRF-GAL4 by co-transfection with constitutively active calmodulin kinase IV and protein kinase A, and binding of Ser(103)-phosphorylated SRF in nuclear extracts, indicated that the SRE.SRF complexes contribute to the Ca(2+)-mediated transcriptional regulation of Egr-1. The results of the current experiments demonstrate for the first time SRE dependent transcription and the role of SRF, a transcription factor known to be a major component of growth responses, in glucose-mediated transcriptional regulation in insulinoma cells. PMID- 10829029 TI - Differential effects of protein kinase C on the levels of epithelial Na+ channel subunit proteins. AB - Regulation of epithelial Na(+) channel (ENaC) subunit levels by protein kinase C (PKC) was investigated in A6 cells. PKC activation altered ENaC subunit levels, differentially decreasing the levels of both beta and gamma, but not alphaENaC. Temporal regulation of beta and gammaENaC by PKC differed; gammaENaC decreased with a time constant of 3.7 +/- 1.0 h, whereas betaENaC decreased in 13.9 +/- 3. 0 h. Activation of PKC also resulted in a decrease in trans-epithelial Na(+) reabsorption for up to 48 h. PMA activation of PKC resulted in negative feedback inhibition of PKC protein levels beginning within 4 h. Both beta and gammaENaC levels, as well as transport tended toward pretreatment values after 48 h of PMA treatment. PKC inhibitors attenuated the effects of PMA on ENaC subunit levels and Na(+) transport. These results directly show for the first time that PKC differentially regulates ENaC subunit levels by decreasing the levels of beta and gamma but not alphaENaC protein. These results imply a PKC-dependent, long term decrease in Na(+) reabsorption. PMID- 10829030 TI - Isolation of a tarantula toxin specific for a class of proton-gated Na+ channels. AB - Acid sensing is associated with nociception, taste transduction, and perception of extracellular pH fluctuations in the brain. Acid sensing is carried out by the simplest class of ligand-gated channels, the family of H(+)-gated Na(+) channels. These channels have recently been cloned and belong to the acid-sensitive ion channel (ASIC) family. Toxins from animal venoms have been essential for studies of voltage-sensitive and ligand-gated ion channels. This paper describes a novel 40-amino acid toxin from tarantula venom, which potently blocks (IC(50) = 0.9 nm) a particular subclass of ASIC channels that are highly expressed in both central nervous system neurons and sensory neurons from dorsal root ganglia. This channel type has properties identical to those described for the homomultimeric assembly of ASIC1a. Homomultimeric assemblies of other members of the ASIC family and heteromultimeric assemblies of ASIC1a with other ASIC subunits are insensitive to the toxin. The new toxin is the first high affinity and highly selective pharmacological agent for this novel class of ionic channels. It will be important for future studies of their physiological and physio-pathological roles. PMID- 10829032 TI - A conserved negatively charged cluster in the active site of creatine kinase is critical for enzymatic activity. AB - Creatine kinase catalyzes the reversible transphosphorylation of creatine by MgATP. From the sequence homology and the molecular structure of creatine kinase isoenzymes, we have identified several highly conserved residues with a potential function in the active site: a negatively charged cluster (Glu(226), Glu(227), Asp(228)) and a serine (Ser(280)). Mutant proteins E226Q, E226L, E227Q, E227L, D228N, and S280A/S280D of human sarcomeric mitochondrial creatine kinase were generated by in vitro mutagenesis, expressed in Escherichia coli, and purified to homogeneity. Their overall structural integrity was confirmed by CD spectroscopy and gel filtration chromatography. The enzymatic activity of all proteins mutated in the negatively charged cluster was extremely low (0.002-0.4% of wild type) and showed apparent Michaelis constants (K(m)) similar to wild type, suggesting that most of the residual activity may be attributed to wild-type revertants. Mutations of Ser(280) led to higher residual activities and altered K(m) values; S280A showed an increase of K(m) for phosphocreatine (65-fold), creatine (6 fold), and ATP (6-fold); S280D showed a decrease of K(m) for creatine (6-fold). These results, together with the transition state structure of the homologous arginine kinase (Zhou, G., Somasundaram, T., Blanc, E., Parthasarathy G., Ellington, W. R., and Chapman, M. S. (1998) Proc. Natl. Acad. Sci. U. S. A. 95, 8449-8454), strongly suggest a critical role of Glu(226), Glu(227), and Asp(228) in substrate binding and catalysis and point to Glu(227) as a catalytic base. PMID- 10829031 TI - Essential role of insulin receptor substrate-2 in insulin stimulation of Glut4 translocation and glucose uptake in brown adipocytes. AB - Insulin and insulin-like growth factor I signals are mediated via phosphorylation of a family of insulin receptor substrate (IRS) proteins, which may serve both complementary and overlapping functions in the cell. To study the metabolic effects of these proteins in more detail, we established brown adipocyte cell lines from wild type and various IRS knockout (KO) animals and characterized insulin action in these cells in vitro. Preadipocytes derived from both wild type and IRS-2 KO mice could be fully differentiated into mature brown adipocytes. In differentiated IRS-2 KO adipocytes, insulin-induced glucose uptake was decreased by 50% compared with their wild type counterparts. This was the result of a decrease in insulin-stimulated Glut4 translocation to the plasma membrane. This decrease in insulin-induced glucose uptake could be partially reconstituted in these cells by retrovirus-mediated re-expression of IRS-2, but not overexpression of IRS-1. Insulin signaling studies revealed a total loss of IRS-2-associated phosphatidylinositol (PI) 3-kinase activity and a reduction in phosphotyrosine associated PI 3-kinase by 30% (p < 0.05) in the KO cells. The phosphorylation and activity of Akt, a major downstream effector of PI 3-kinase, as well as Akt dependent phosphorylation of glycogen synthase kinase-3 and p70S6 kinase were not affected by the lack of IRS-2; however, there was a decrease in insulin stimulation of Akt associated with the plasma membrane. These results provide evidence for a critical role of IRS-2 as a mediator of insulin-stimulated Glut4 translocation and glucose uptake in adipocytes. This occurs without effects in differentiation, total activation of Akt and its downstream effectors, but may be caused by alterations in compartmentalization of these downstream signals. PMID- 10829033 TI - A maize homologue of the bacterial CMP-3-deoxy-D-manno-2-octulosonate (KDO) synthetases. Similar pathways operate in plants and bacteria for the activation of KDO prior to its incorporation into outer cellular envelopes. AB - The eight-carbon acid sugar 3-deoxy-d-manno-2-octulosonate (KDO) is an essential component of Gram-negative bacterial cell walls and capsular polysaccharides. KDO is incorporated into these polymers as CMP-KDO, which is produced in an unusual activation step catalyzed by the enzyme CMP-KDO synthetase. CMP-KDO synthetase activity has traditionally been considered exclusive to Gram-negative bacteria. CMP-KDO synthetase inhibitors attract great interest owing to their potential as selective bactericides. The sugar KDO is also a component of the rhamnogalacturonan II pectin fraction of the primary cell walls of most higher plants and of the cell wall polysaccharides of some green algae. However, the metabolic pathway leading to its incorporation into the plant cell wall is unknown. This paper describes the isolation and characterization of a maize gene, which codes for a protein very similar in sequence and activity to prokaryotic CMP-KDO synthetases. Remarkably, the maize gene can complement a CMP-KDO synthetase (kdsB) Salmonella typhimurium mutant defective in cell wall synthesis. ZmCKS activity is novel in eukaryotes. The evolutionary origin of ZmCKS is discussed in relation to the high degree of conservation between the plant and bacterial genes and its atypical codon usage in maize. PMID- 10829034 TI - The cAMP-specific phosphodiesterase PDE4A5 is cleaved downstream of its SH3 interaction domain by caspase-3. Consequences for altered intracellular distribution. AB - The unique N-terminal region of the cAMP-specific phosphodiesterase PDE4A5, which confers an ability to bind to certain protein SH3 domains, is cleaved during apoptosis in both Rat-1 fibroblasts and PC12 cells. Cleavage was abolished by the caspase-3-selective inhibitor, z-DEVD-CHO but not the caspase-1 selective inhibitor, z-YVAD-CHO. Caspase-3 treatment of PDE4A5, expressed either transiently in COS cells or generated in vitro by coupled transcription translation, generated a similar cleavage product of 100 kDa compared with the native 110-kDa PDE4A5. This product could be detected immunochemically with an antibody raised to a C-terminal PDE4A5 peptide but not an antibody raised to the N terminus of PDE4A5, indicating that caspase-3 caused N-terminal cleavage of PDE4A5. Deletion of the putative caspase-3 cleavage site, (69)DAVD(72), in PDE4A5, or generation of either the D72A or the D69A mutants, ablated the ability of caspase-3 to cause cleavage. The N-terminal truncate PDE4A5-DeltaP3 was engineered to mimic the caspase-cleaved product of PDE4A5. This showed altered catalytic activity and, unlike PDE4A5, was unable to interact with the SH3 domain of the tyrosyl kinase, LYN. Although both PDE4A5 and PDE4A5-DeltaP3 were localized at cell cortical regions (ruffles), the distinct perinuclear association noted for both PDE4A5 and LYN was not seen for PDE4A5-DeltaP3. Staurosporine-induced apoptosis caused a marked redistribution of PDE4A5 but not PDE4A8 in stably transfected Rat-1 cells. The PDE4-selective inhibitor, rolipram together with the adenylyl cyclase activator forskolin, caused a synergistic increase in the apoptosis of Rat-1 cells. Overexpression of PDE4A5 in Rat-1 cells protected against staurosporine-induced apoptosis in contrast to overexpression of PDE4A8, which potentiated apoptosis. PDE4A5 may be the sole PDE4 family member to provide a substrate for caspase-3 cleavage and this action serves to remove the SH3 binding domain that is unique to this isoform within the PDE4A family and to alter its intracellular targeting. PMID- 10829035 TI - A novel electrogenic amino acid transporter is activated by K+ or Na+, is alkaline pH-dependent, and is Cl--independent. AB - A new eukaryotic nutrient amino acid transporter has been cloned from an epithelium that is exposed to high voltages and alkaline pH. The full-length cDNA encoding this novel CAATCH1 (cation-anion-activated Amino acid transporter/channel) was isolated using a polymerase chain reaction-based strategy, and its expression product in Xenopus oocytes displayed a combination of several unique, unanticipated functional properties. CAATCH1 electrophysiological properties resembled those of Na(+),Cl(-)-coupled neurotransmitter amine transporters, although CAATCH1 was cloned from a gut absorptive epithelium rather than from an excitable tissue. Amino acids such as l proline, l-threonine, and l-methionine elicited complex current-voltage relationships in alkaline pH-dependent CAATCH1 that were reminiscent of the behavior of the dopamine, serotonin, and norepinephrine transporters (DAT, SERT, NET) in the presence of their substrates and pharmacological inhibitors such as cocaine or antidepressants. These I-V relationships indicated a combination of substrate-associated carrier current plus an independent CAATCH1-associated leakage current that could be blocked by certain amino acids. However, unlike all structurally related proteins, CAATCH1 activity is absolutely independent of Cl( ). Unlike related KAAT1, CAATCH1 possesses a methionine-inhibitable constitutive leakage current and is able to switch its narrow substrate selectivity, preferring threonine in the presence of K(+) but preferring proline in the presence of Na(+). PMID- 10829036 TI - Molecular basis for differential substrate specificity in class IV alcohol dehydrogenases: a conserved function in retinoid metabolism but not in ethanol oxidation. AB - Mammalian class IV alcohol dehydrogenase enzymes are characteristic of epithelial tissues, exhibit moderate to high K(m) values for ethanol, and are very active in retinol oxidation. The human enzyme shows a K(m) value for ethanol which is 2 orders of magnitude lower than that of rat class IV. The uniquely significant difference in the substrate-binding pocket between the two enzymes appears to be at position 294, Val in the human enzyme and Ala in the rat enzyme. Moreover, a deletion at position 117 (Gly in class I) has been pointed out as probably responsible for class IV specificity toward retinoids. With the aim of establishing the role of these residues, we have studied the kinetics of the recombinant human and rat wild-type enzymes, the human G117ins and V294A mutants, and the rat A294V mutant toward aliphatic alcohols and retinoids. 9-cis-Retinol was the best retinoid substrate for both human and rat class IV, strongly supporting a role of class IV in the generation of 9-cis-retinoic acid. In contrast, 13-cis retinoids were not substrates. The G117ins mutant showed a decreased catalytic efficiency toward retinoids and toward three-carbon and longer primary aliphatic alcohols, a behavior that resembles that of the human class I enzyme, which has Gly(117). The K(m) values for ethanol dramatically changed in the 294 mutants, where the human V294A mutant showed a 280-fold increase, and the rat A294V mutant a 50-fold decrease, compared with those of the respective wild-type enzymes. This demonstrates that the Val/Ala exchange at position 294 is mostly responsible for the kinetic differences with ethanol between the human and rat class IV. In contrast, the kinetics toward retinoids was only slightly affected by the mutations at position 294, compatible with a more conserved function of mammalian class IV alcohol dehydrogenase in retinoid metabolism. PMID- 10829037 TI - Hereditary nonpolyposis colorectal cancer: a call for attention. PMID- 10829038 TI - Population-based molecular detection of hereditary nonpolyposis colorectal cancer. AB - PURPOSE: Cancer morbidity and mortality can be dramatically reduced by colonoscopic screening of individuals with the hereditary nonpolyposis colorectal cancer (HNPCC) syndrome, creating a need to identify HNPCC. We studied how HNPCC identification should be carried out on a large scale in a sensitive and efficient manner. PATIENTS AND METHODS: Colorectal cancer specimens from consecutive newly diagnosed patients were studied for microsatellite instability (MSI). Germline mutations in the MLH1 and MSH2 genes were searched for in MSI(+) individuals. RESULTS: Among 535 colorectal cancer patients, 66 (12%) were MSI(+). Among these, 18 (3.4% of the total) had disease-causing germline mutations in MLH1 or MSH2. Among these 18 patients, five were less than 50 years old, seven had a previous or synchronous colorectal or endometrial cancer, and 15 had at least one first-degree relative with colorectal or endometrial cancer. Notably, 17 (94%) of 18 patients had at least one of these three features, which were present in 22% of all 535 patients. Combining these data with a previous study of 509 patients, mutation-positive HNPCC accounts for 28 (2.7%) of 1,044 cases of colorectal cancer, predicting a greater than one in 740 incidence of mutation positive individuals in this population. CONCLUSION: Large-scale molecular screening for HNPCC can be done by the described two-stage procedure of MSI determination followed by mutation analysis. Efficiency can be greatly improved by using three high-risk features to select 22% of all patients for MSI analysis, whereby only 6% need to have mutation analysis. Sensitivity is only slightly impaired by this procedure. PMID- 10829039 TI - c-erbB-2 is of independent prognostic relevance in gastric cancer and is associated with the expression of tumor-associated protease systems. AB - PURPOSE: The c-erbB-2 gene (encoding the protein p185) is overexpressed in diverse human cancers and has been implicated to be of prognostic value in gastric cancer. Recent studies suggest a role of p185 in tumor progression by specifically promoting the invasive capacity of tumor cells. Therefore, the present study was conducted with the following three objectives: (1) to support the prognostic value of c-erbB-2 in gastric cancer in a large prospective series using a monoclonal antibody and a highly sensitive immunohistochemical method; (2) to determine the association of c-erbB-2 expression with the expression of invasion-related genes; and (3) to perform the first overall multivariate analysis including c-erbB-2 and the invasion-related tumor-associated protease systems. PATIENTS AND METHODS: In a consecutive prospective series of 203 gastric cancer patients (median follow-up, 42 months), expression of c-erbB-2 and a panel of tumor-associated proteases and inhibitors by tumor cells were evaluated semiquantitatively (score 0 to 3) and analyzed for correlation (chi(2) test, Bonferroni-corrected). Kaplan-Meier survival analysis and multivariate Cox analysis were performed to determine the relative prognostic impact of c-erbB-2 and the invasion-related parameters. RESULTS: Kaplan-Meier analysis (log-rank statistics) revealed a significant association of increasing expression of c-erbB 2 with shorter disease-free (P =. 0023) and overall survival (P =.0160). High amounts of p185 were significantly associated with a high expression of urokinase type plasminogen activator (uPA) (P <.010), uPA-receptor (P =.030), type-1 plasminogen activator inhibitor (PAI) (P <.010), type-2 PAI (P =.021), cathepsin D (P =.036), matrix metalloproteinase-2 (P =. 024), alpha-1-antichymotrypsin (P =.025), and alpha-2-macroglobulin (P =.017). Multivariate analysis considering these proteases/protease inhibitors, in addition to alpha-1-antitrypsin, tissue plasminogen activator, plasminogen, alpha-2-antiplasmin, and antithrombin III, and established prognostic parameters revealed that, in addition to surgical curability, pT stage, pN stage, and PAI-1, c-erbB-2 is an independent prognostic factor for overall survival of curatively resected patients (n = 139; P =.049; relative risk, 1.54; 95% confidence interval, 1.08 to 1.67) and all patients (P =.028; relative risk 1.33; 95% CI, 1.28 to 1.38). CONCLUSION: c-erbB-2 is confirmed as a new independent, functional prognostic parameter for overall survival in gastric cancer, even when a panel of invasion-related factors, including the strong prognostic parameter PAI-1, are considered. The significant correlation of p185 with several tumor-associated proteases supports the hypothesis that c-erbB-2 is a promoter of invasion and metastasis. This strongly suggests that c-erbB-2 may be a promising target for anti-invasive therapy in gastric cancer. PMID- 10829040 TI - Escalated focal liver radiation and concurrent hepatic artery fluorodeoxyuridine for unresectable intrahepatic malignancies. AB - PURPOSE: To evaluate the response, time to progression, survival, and impact of radiation (RT) dose on survival in patients with intrahepatic malignancies treated on a phase I trial of escalated focal liver RT. PATIENTS AND METHODS: From April 1996 to January 1998, 43 patients with unresectable intrahepatic hepatobiliary cancer (HB; 27 patients) and colorectal liver metastases (LM; 16 patients) were treated with high-dose conformal RT. The median tumor size was 10 x 10 x 8 cm. The median RT dose was 58.5 Gy (range, 28.5 to 90 Gy), 1.5 Gy twice daily, with concurrent continuous-infusion hepatic arterial fluorodeoxyuridine (0.2 mg/kg/d) during the first 4 weeks of RT. RESULTS: The response rate in 25 assessable patients was 68% (16 partial and one complete response). With a median potential follow-up period of 26.5 months, the median times to progression for all tumors, LM, and HB were 6, 8, and 3 months, respectively. The median survival times of all patients, patients with LM, and patients with HB were 16, 18, and 11 months, respectively. On multivariate analyses, escalated RT dose was independently associated with improved progression-free and overall survival. The median survival of patients treated with 70 Gy or more has not yet been reached (16.4+ months), compared with 11.6 months in patients treated with lower RT doses (P =.0003). CONCLUSION: The excellent response rate, prolonged intrahepatic control, and improved survival in patients treated with RT doses of 70 Gy or more motivate continuation of dose-escalation studies for patients with intrahepatic malignancies. PMID- 10829041 TI - Radiation therapy for squamous cell carcinoma of the tonsillar region: a preferred alternative to surgery? AB - PURPOSE: There are no definitive randomized studies that compare radiotherapy (RT) with surgery for tonsillar cancer. The purpose of this study was to evaluate the results of RT alone and RT combined with a planned neck dissection for carcinoma of the tonsillar area and to compare these data with the results of treatment with primary surgery. PATIENTS AND METHODS: Four hundred patients were treated between October 1964 and December 1997 and observed for at least 2 years. One hundred forty-one patients underwent planned neck dissection, and 18 patients received induction (17 patients) or concomitant (one patient) chemotherapy. RESULTS: Five-year local control rates, by tumor stage, were as follows: T1, 83%; T2, 81%; T3, 74%; and T4, 60%. Multivariate analysis revealed that local control was significantly influenced by tumor stage (P =.0001), fractionation schedule (P =.0038), and external beam dose (P =.0227). Local control after RT for early stage cancers was higher for tonsillar fossa/posterior pillar cancers than for those arising from the anterior tonsillar pillar. Five-year cause-specific survival rates, by disease stage, were as follows: I, 100%; II, 86%; III, 82%; IVa, 63%; and IVb, 22%. Multivariate analysis revealed that cause-specific survival was significantly influenced by overall stage (P =.0001), planned neck dissection (P =.0074), and histologic differentiation (P =.0307). The incidence of severe late complications after treatment was 5%. CONCLUSION: RT alone or combined with a planned neck dissection provides cure rates that are as good as those after surgery and is associated with a lower rate of severe complications. PMID- 10829042 TI - Subcutaneous administration of amifostine during fractionated radiotherapy: a randomized phase II study. AB - PURPOSE: Amifostine (WR-2721) is an important cytoprotective agent. Although intravenous administration is the standard route, pharmacokinetic studies have shown acceptable plasma levels of the active metabolite of amifostine (WR-1605) after subcutaneous administration. The subcutaneous route, due to its simplicity, presents multiple advantages over the intravenous route when amifostine is used during fractionated radiotherapy. PATIENTS AND METHODS: Sixty patients with thoracic, 40 with head and neck, and 40 with pelvic tumors who were undergoing radical radiotherapy were enrolled onto a randomized phase II trial to assess the feasibility, tolerance, and cytoprotective efficacy of amifostine administered subcutaneously. A flat dose of amifostine 500 mg, diluted in 2.5 mL of normal saline, was injected subcutaneously 20 minutes before each radiotherapy fraction. RESULTS: The subcutaneous amifostine regimen was well tolerated by 85% of patients. In approximately 5% of patients, amifostine therapy was interrupted due to cumulative asthenia, and in 10%, due to a fever/rash reaction. Hypotension was never noted, whereas nausea was frequent. A significant reduction of pharyngeal, esophageal, and rectal mucositis was noted in the amifostine arm (P <.04). The delays in radiotherapy because of grade 3 mucositis were significantly longer in the group of patients treated with radiotherapy alone (P <.04). Amifostine significantly reduced the incidence of acute perineal skin and bladder toxicity (P <.0006). CONCLUSION: Subcutaneous administration of amifostine is well tolerated, effectively reduces radiotherapy's early toxicity, and prevents delays in radiotherapy. The subcutaneous route is much simpler and saves time compared with the intravenous route of administration and can be safely and effectively applied in the daily, busy radiotherapy practice. PMID- 10829043 TI - Activity of exemestane in metastatic breast cancer after failure of nonsteroidal aromatase inhibitors: a phase II trial. AB - PURPOSE: To evaluate the antitumor activity and toxicity of a new steroidal aromatase inactivator, exemestane, in postmenopausal women with metastatic breast cancer who had progressive disease (PD) after treatment with a nonsteroidal aromatase inhibitor. PATIENTS AND METHODS: In this phase II trial, eligible patients were treated with exemestane 25 mg daily (n = 241) followed, at the time PD was determined, by exemestane 100 mg daily (n = 58). RESULTS: On the basis of the intent-to-treat analysis by independent review, exemestane 25 mg produced objective responses in 6.6% of patients (95% confidence interval [CI], 3.8% to 10.6%) and overall success (complete response + partial response + no change for 24 weeks or longer) in 24.3% (95% CI, 19.0% to 30.2%). The median durations of objective response and overall success were 58.4 weeks (95% CI, 49.7 to 71.1 weeks) and 37.0 weeks (95% CI, 35.0 to 39.4 weeks), respectively. Increasing the dose of exemestane to 100 mg upon the development of PD produced one partial response (1.7%; 95% CI, 0.0% to 9.2%). Both dosages were well tolerated and were discontinued because of adverse events in only 1.7% of patients. CONCLUSION: Exemestane 25 mg once daily seems to be an attractive alternative to chemotherapy for the treatment of patients with metastatic breast cancer after multiple hormonal therapies have failed. PMID- 10829044 TI - Gemcitabine plus cisplatin repeating doublet therapy in previously treated, relapsed breast cancer patients. AB - PURPOSE: To determine the safety and efficacy of gemcitabine plus cisplatin for patients with relapsed adenocarcinoma of the breast. PATIENTS AND METHODS: Previously treated patients with adenocarcinoma of the breast received cisplatin (30 mg/m(2)) plus gemcitabine (1,000 mg/m(2)) on days 1, 8, and 15 of each 28-day cycle, which was changed after patient no. 12 to cisplatin (30 mg/m(2)) plus gemcitabine (750 mg/m(2)) days 1 and 8 of each 21-day cycle. RESULTS: Of 30 patients, three (10%) had complete and 12 (40%) had partial responses, for an overall response rate of 50%. Two objective responses were observed among the four patients accrued after relapse that followed high-dose/stem-cell therapies. The median time to progression was 14 weeks. The median time to progression for objective responders was 23.5 weeks, with a range of 8 to 68 weeks. Toxicities included grades III and IV neutropenia in 13%, anemia in 6%, thrombocytopenia in 31%, grade III nausea in 4%, and grade II peripheral neuropathy in 2% of 151 treatment cycles. Moderate alopecia occurred in four patients. There were no treatment-related deaths. CONCLUSION: Cisplatin plus gemcitabine is active and tolerable for patients with relapsed breast cancer. Responses observed in previously treated patients, including high-dose/stem-cell failures, indicate activity in otherwise drug-refractory patients. PMID- 10829045 TI - Intention to undergo prophylactic bilateral mastectomy in women at increased risk of developing hereditary breast cancer. AB - PURPOSE: To assess intention to undergo prophylactic bilateral mastectomy and psychologic determinants in unaffected women at increased risk of developing hereditary breast cancer. PATIENTS AND METHODS: Three hundred thirty-three women who were awaiting their initial appointments for risk assessment, advice about surveillance, and prophylactic options at one of 14 familial cancer clinics participated in a cross-sectional, questionnaire-based survey. RESULTS: Nineteen percent of women would consider and 47% would not consider a prophylactic mastectomy, should genetic testing identify a mutation in a breast cancer predisposing gene, whereas 34% were unsure and 1% had already undergone a prophylactic mastectomy. In a bivariate analysis, women at a moderately increased risk of developing breast cancer had the highest proportion of subjects reporting that they would consider a prophylactic mastectomy (25%), compared with women at high risk (16%) (chi(2) = 7.79; P =.051). In multivariate analyses, consideration of prophylactic mastectomy strongly correlated with high levels of breast cancer anxiety (odds ratio [OR] = 17.4; 95% confidence interval [CI], 4.35 to 69.71; P =. 0001) and overestimation of one's breast cancer risk (OR = 3.01; 95% CI, 1.43 to 6.32; P =.0036), whereas there was no association with objective breast cancer risk (P =.60). CONCLUSION: A significant proportion of women at increased risk of developing hereditary breast cancer would consider prophylactic mastectomy. Although prophylactic mastectomy may be appropriate in women at high risk of developing breast cancer, it is perhaps less so in those who have a moderately increased risk. Such moderate-risk women are likely to benefit from interventions aimed at reducing breast cancer anxiety and correcting exaggerated breast cancer risk perceptions. PMID- 10829046 TI - Declining cancer rates in the 1990s. AB - PURPOSE: To provide evidence of a substantial decline in cancer rates for the period 1991 through 1995 and characterize major risk factors that seem to be driving secular trends in cancer mortality and incidence. DESIGN: Incidence and mortality rates were calculated using national surveillance data collected through the Surveillance, Epidemiology, and End Results (SEER) program and the National Center for Health Statistics. RESULTS: All-sites cancer incidence and mortality fell in the period 1991 through 1995; this decline is largely attributable to decreases in the smoking-related cancers, especially lung cancer. Of the 20 leading incident cancers today, both incidence and mortality are decreasing among 11 sites for men and 12 for women. In men, the decline in mortality has been notable and is especially apparent for the smoking-related cancers, including those of the lung, oral cavity and pharynx, larynx, and, to a lesser extent, bladder. In women, all-sites mortality decreased only approximately 0.4% from 1991 through 1995. Three cancers continued to show substantial increases in mortality through 1995 for both men and women (liver, multiple myeloma, and non-Hodgkin's lymphoma), while incidence rates continued to climb for liver cancer, non-Hodgkin's lymphoma, and melanoma. CONCLUSION: Data from the SEER program on recent trends in cancer incidence and mortality show that cancer rates are generally on the decline, largely because of reductions in smoking-related cancers. A consistent increase in mortality rates due to liver cancer poses a new health care challenge, one that will require the development of an effective treatment for individuals currently infected with hepatitis C or B to prevent mortality rates from continuing to increase. PMID- 10829047 TI - Filgrastim as an alternative to donor leukocyte infusion for relapse after allogeneic stem-cell transplantation. AB - PURPOSE: Donor leukocyte infusion (DLI) effectively treats relapse after allogeneic stem-cell transplantation (alloSCT), but the response may require several months and may be associated with significant toxicity. Filgrastim has also been observed to effectively treat leukemic relapse after alloSCT. A retrospective analysis was performed to determine the effectiveness of filgrastim in treating relapses after alloSCT. PATIENTS AND METHODS: Fourteen patients with hematologic malignancies were treated with filgrastim at relapse after alloSCT. Filgrastim was given at 5 mcg/kg/d subcutaneously for 21 consecutive days. Response was evaluated at 7 days after completion of filgrastim. Immunosuppressants, if present, were rapidly tapered to complete discontinuation at the time of relapse. RESULTS: Three patients were not assessable for response because additional therapy was necessary before completion of filgrastim. Six patients (43%) achieved a complete response on an intent-to-treat basis. When response was evaluated based on relapse type, three of four cytogenetic relapses, two of three morphologic relapses, and one of four hematologic relapses achieved a complete remission. Two responses were observed in patients who were completely off of any immunosuppression at the time of relapse. Six patients developed chronic graft-versus-host disease. The event-free and overall survival rates for all 14 patients are 43% and 73%, respectively. CONCLUSION: The use of filgrastim with rapid discontinuation of immunosuppression results in response rates that are similar to results using DLI. Filgrastim could be considered as an alternative or an adjunct to DLI for relapses after alloSCT. PMID- 10829048 TI - Molecular remission after allogeneic or autologous transplantation of hematopoietic stem cells for multiple myeloma. AB - PURPOSE: To assess the clinical relevance of minimal residual disease (MRD) in patients with multiple myeloma (MM), 50 patients were monitored while they were in complete clinical remission (CCR) after autologous or allogeneic stem-cell transplantation. PATIENTS AND METHODS: Stringent molecular monitoring using clonal markers based on rearranged immunoglobulin heavy-chain genes was performed in 44 of 50 MM patients in CCR. Molecular clinical remission (MCR) was defined as more than one consecutive negative polymerase chain reaction (PCR) test result. RESULTS: Twelve (27%) of 44 molecularly monitored patients achieved MCR; four of the 12 became PCR-positive, and one of these four relapsed. In comparison with patients who did not achieve MCR, patients who achieved MCR had a significantly lower relapse rate (41% v 16%; P <.05) and longer relapse-free survival (35 v 110 months; P <.005). Fourteen of 26 patients in CCR who had received allografts were evaluated on a molecular basis: seven (50%) of the 14 achieved MCR and did not relapse; one of the seven remaining patients relapsed. Thirty of 47 patients in CCR who received autografts were evaluated on a molecular basis: five (16%) of the 30 achieved MCR; two of these five became PCR-negative, and one of these two relapsed. Ten of the 25 remaining patients later relapsed. For these nonrandomized groups, the higher MCR rate after allograft procedures was statistically significant (P <.01; Fisher's exact test). CONCLUSION: MCR can be obtained in a relatively high proportion of MM patients who have achieved CCR after undergoing allograft procedures and in a smaller fraction of patients after undergoing autograft procedures. In approximately one fourth of MM patients who achieve CCR after transplantation, it may be possible to keep the disease burden constantly below the PCR threshold. Because MCR was associated with prolonged relapse-free survival, these patients could have a relatively favorable clinical outcome. PMID- 10829049 TI - Phase I trial of the anti-Lewis Y drug immunoconjugate BR96-doxorubicin in patients with lewis Y-expressing epithelial tumors. AB - PURPOSE: We conducted a phase I clinical trial of BR96-Doxorubicin (BR96-Dox), a chimeric anti-Lewis Y (Le(Y)) monoclonal antibody conjugated to doxorubicin, in patients whose tumors expressed the Le(Y) antigen. The study aimed to determine the toxicity, maximum-tolerated dose, pharmacokinetics, and immunogenicity of BR96-Dox. PATIENTS AND METHODS: This was a phase I dose escalation study. BR96 Dox was initially administered alone as a 2-hour infusion every 3 weeks. The occurrence of gastrointestinal (GI) toxicity necessitated the administration of BR96-Dox as a continuous infusion over 24 hours and use of antiemetics and antigastritis premedication. Patients experiencing severe GI toxicity underwent GI endoscopy. All patients underwent restaging after two cycles. RESULTS: A total of 66 patients predominantly with metastatic colon and breast cancer were enrolled onto the study. The most common side effects were GI toxicity, fever, and elevation of pancreatic lipase. At higher doses, BR96-Dox was associated with nausea, vomiting, and endoscopically documented exudative gastritis of the upper GI tract, which was dose-limiting at a maximum dose of 875 mg/m(2) (doxorubicin equivalent, 25 mg/m(2)) administered every 3 weeks. Toxicity was reversible and generally of short duration. Premedication with the antiemetic Kytril (granisetron hydrochloride; SmithKline Beecham, Philadelphia, PA), the antacid omeprazole, and dexamethasone was most effective in ameliorating GI toxicity. A dose of 700 mg/m(2) BR96-Dox (doxorubicin equivalent, 19 mg/m(2)) every 3 weeks was determined to be the optimal phase II dose when administered with antiemetic and antigastritis prophylaxis. BR96-Dox deposition on tumor tissue was documented immunohistochemically and by confocal microscopy. At the 550-mg/m(2) dose, the half-life (mean +/- SD) of BR96 and doxorubicin was 300 +/- 95 hours and 43 +/- 4 hours, respectively. BR96-Dox elicited a weak immune response in 37% of patients. Objective clinical responses were seen in two patients. CONCLUSION: BR96-Dox provides a unique strategy to deliver doxorubicin to Le(Y)-expressing tumor and was well tolerated at doses of 700 mg/m(2) every 3 weeks. BR96-Dox was not associated with the typical side-effect profile of native doxorubicin and can potentially deliver high doses of doxorubicin to antigen-expressing tumors. A phase II study in doxorubicin-sensitive tumors is warranted. PMID- 10829050 TI - Phase I, dose-finding, and pharmacokinetic study of raltitrexed combined with oxaliplatin in patients with advanced cancer. AB - PURPOSE: To determine the maximum-tolerated dose (MTD) and the dose-limiting toxicities (DLTs) of the raltitrexed plus oxaliplatin combination regimen, to explore its safety and pharmacokinetics, and to assess its antitumor activity in patients with advanced solid tumors. PATIENTS AND METHODS: Forty-eight patients received the combination of raltitrexed plus oxaliplatin. Raltitrexed was administered as a 15-minute infusion followed by oxaliplatin as a 2-hour infusion 1 hour later, repeated every 3 weeks. Seven dose levels were explored, ranging from 2 to 3.75 mg/m(2) and from 85 to 130 mg/m(2) for raltitrexed and oxaliplatin, respectively. The pharmacokinetics of both raltitrexed and oxaliplatin was assessed at the last three dose levels. RESULTS: Forty-six patients were assessable for toxicity. Severe toxicities usually occurred from dose level V (raltitrexed 3 mg/m(2) and oxaliplatin 130 mg/m(2)). This combination was not myelosuppressive, eliciting only sporadic grades 3 and 4 neutropenia and/or thrombocytopenia without complications. There was no alopecia. DLTs were asthenia and nausea/vomiting, despite systematic antiemetic prophylaxis. Dose level VI (raltitrexed 3.5 mg/m(2) and oxaliplatin 130 mg/m(2)) was deemed to be the MTD. Eight confirmed partial responses were observed: six patients with malignant mesothelioma (both pretreated and nonpretreated), one with fluorouracil-refractory pancreatic carcinoma, and one with renal carcinoma. Evaluation of the pharmacokinetics of both drugs did not suggest any drug interaction. CONCLUSION: The combination of raltitrexed and oxaliplatin given as consecutive short infusions every 3 weeks seems to be an acceptable regimen that allows a dose-intensity as high as the sum of the recommended doses of each agent given alone. The dose recommended for further phase II studies is raltitrexed 3 mg/m(2) and oxaliplatin 130 mg/m(2) every 3 weeks. Promising antitumor activity has been observed in patients with malignant mesothelioma. PMID- 10829051 TI - Correlation between docetaxel clearance and estimated cytochrome P450 activity by urinary metabolite of exogenous cortisol. AB - PURPOSE: There is no simple and practical method for the estimation of the interpatient variability of cytochrome P450 (CYP3A4) enzyme activity. Cortisol is metabolized by this enzyme and excreted as 6-beta-hydroxycortisol (6beta-OHF) and free-cortisol (FC) in urine, and docetaxel is also metabolized by hepatic CYP3A4 enzyme. We developed a new method for the estimation of CYP3A4 activity by measuring urinary cortisol metabolite after administration of exogenous cortisol. This study was aimed at assessing the predictability of the individual activity of CYP3A4 estimated by this method. PATIENTS AND METHODS: Thirty patients with advanced non-small-cell lung cancer were entered onto this study. Hydrocortisone 300 mg was administered intravenously, and urinary 6beta-OHF and FC were measured. More than 2 days later, 60 mg/m(2) of docetaxel was administered intravenously with pharmacokinetic sampling. The correlation between docetaxel pharmacokinetics and estimated interpatient variability of CYP3A4 activity with the application of our method was assessed. RESULTS: After cortisol administration, the total amount of 24-hour urinary 6beta-OHF (T6beta-OHF) increased about 60-fold compared with pretreatment levels, averaging 12,273 +/- 4,076 microg/d (mean +/- SD). Docetaxel clearance (CL) and area under the concentration-time curve averaged 24.5 +/- 6.4 L/h/m(2) and 2.66 +/- 0.91 mg/L 8729. h, respectively. An excellent correlation between docetaxel CL and T6beta OHF was observed (r =.867). In multivariate analysis, T6beta-OHF (P <.001), alpha 1-acid glycoprotein (P <.004), AST (P =.007), and age (P =. 022) significantly correlated with docetaxel CL. CONCLUSION: The interpatient variability of CYP3A4 activity and docetaxel CL could be predicted by measuring T6beta-OHF after cortisol administration. PMID- 10829052 TI - Cancer risk and low-penetrance susceptibility genes in gene-environment interactions. AB - PURPOSE: To provide a concise review for human cancer risk related to low penetrance genes and their effects on environmental carcinogen exposure. METHODS: Citation of relevant and recent references for molecular epidemiology, focusing on lung cancer, ethical issues, and some clinical implications of recent molecular epidemiology studies. RESULTS: Low-penetrance genes contribute to cancer risk by augmenting the effects of carcinogen exposures. These exposures can be measured in the body through molecular dosimetry (ie, the amount of DNA damage), which reflects a biologically effective dose. The examination of tumors and the mutations within tumor suppressor genes, such as p53, can provide etiologic clues for both exposure and susceptibility. Although many studies have focused on carcinogen metabolism and cancer risk, more recent studies are considering DNA repair. Also, we are learning that behavior, such as tobacco addiction, also may be genetically controlled. CONCLUSION: Sporadic cancers are caused by gene(n)-environment(n) interactions rather than a dominant effect by a specific gene, environmental exposure, or gene-environment interaction. New paradigms, where we categorize genes as caretaker or gatekeeper genes, will allow for new hypotheses to be tested and will require advanced methods of analysis. The goal of molecular epidemiology is to develop risk assessment models for individuals, but currently the most achievable goal will be population risk assessment and a better understanding of carcinogenesis. PMID- 10829053 TI - Psychiatric side effects of interferon therapy: prevalence, proposed mechanisms, and future directions. AB - The increasing use of interferon (IFN) in treating a variety of disorders including, malignant melanoma and hepatitis C, has resulted in the identification and increasing concern about the psychiatric side effects that can result from treatment. These effects can occur either shortly after beginning IFN therapy or later as a result of continued treatment. Studies have reported the incidence of later side effects, which include symptoms of depression, anxiety, and occasional suicidal ideation, to be from 0% to 70%. Case studies have demonstrated that pharmacologic interventions are beneficial in reducing iatrogenic psychiatric symptoms while allowing patients to maintain IFN therapy. The present article provides an overview of the psychiatric effects of IFN therapy, the proposed mechanisms of these side effects, and case studies that provide mechanistic support. In addition, limitations of the current literature are provided with suggestions for treating physicians and a discussion of possible future research directions. PMID- 10829054 TI - Hospital and physician volume or specialization and outcomes in cancer treatment: importance in quality of cancer care. AB - PURPOSE: To conduct a comprehensive review of the health services literature to search for evidence that hospital or physician volume or specialty affects the outcome of cancer care. METHODS: We reviewed the 1988 to 1999 MEDLINE literature that considered the hypothesis that higher volume or specialization equals better outcome in processes or outcomes of cancer treatments. RESULTS: An extensive, consistent literature that supported a volume-outcome relationship was found for cancers treated with technologically complex surgical procedures, eg, most intra abdominal and lung cancers. These studies predominantly measured in-hospital or 30-day mortality and used the hospital as the unit of analysis. For cancer primarily treated with low-risk surgery, there were fewer studies. An association with hospital and surgeon volume in colon cancer varied with the volume threshold. For breast cancer, British studies found that physician specialty and volume were associated with improved long-term outcomes, and the single American report showed an association between hospital volume of initial surgery and better 5-year survival. Studies of nonsurgical cancers, principally lymphomas and testicular cancer, were few but consistently showed better long-term outcomes associated with larger hospital volume or specialty focus. Studies in recurrent or metastatic cancer were absent. Across studies, the absolute benefit from care at high-volume centers exceeds the benefit from break-through treatments. CONCLUSION: Although these reports are all retrospective, rely on registries with dated data, rarely have predefined hypotheses, and may have publication and self interest biases, most support a positive volume-outcome relationship in initial cancer treatment. Given the public fear of cancer, its well-defined first identification, and the tumor-node-metastasis taxonomy, actual cancer care should and can be prospectively measured, assessed, and benchmarked. The literature suggests that, for all forms of cancer, efforts to concentrate its initial care would be appropriate. PMID- 10829055 TI - Two patients with sarcoma. Case 1. Synovial cell sarcoma of the lung. PMID- 10829056 TI - Two patients with sarcoma. Case 2. Uterine sarcoma. PMID- 10829057 TI - Doc, shouldn't we be getting some tests? PMID- 10829058 TI - Leukemia in Mexican versus Mexican-American children. PMID- 10829059 TI - Trastuzumab in CSF. PMID- 10829060 TI - Paclitaxel and cisplatin in ovarian cancer. PMID- 10829061 TI - Different sensitivity to receptor editing of B cells from mice hemizygous or homozygous for targeted Ig transgenes. AB - Ig knock-in mice have been used to study the relative contribution of receptor selection versus clonal selection in the control of autoreactive B cells. The anti-MHC class I 3-83Ig knock-in (3-83Igi) mice manifest extensive receptor editing in the presence of H-2(b). However, receptor editing is also observed on the H-2(d) background, although reactivity toward this antigen is below detection and its presence does not affect the generation of 3-83Ig(+) mature B cells in classical 3-83Ig transgenic mice. In this study we have analyzed the contribution of genetic background, B cell receptor signaling, and transgene copy number on the initiation and extent of receptor editing in the 3-83Igi;H-2(d) mice. Crossing the 3-83Ig insertion into either CD45-deficient H-2(d) mice or onto the BALB/c background reduces the extent of receptor editing and increases the fraction of 3-83Ig-expressing B cells, indicating that in the original line editing depends on B cell receptor signaling induced by cross-reacting antigen(s). However, receptor editing is still detectable in hemizygous 3-83Igi mice even on the BALB/c background, on which the 3-83 antibody was originally raised, whereas it is abrogated in homozygous 3-83Igi;H-2(d) animals. This latter observation indicates that immature B cells expressing immunoglobulin from single heavy and light chain loci, as they do physiologically, utilize receptor editing for an exquisite quality control of their antigen receptor that may only partly be based on self-reactivity. PMID- 10829062 TI - The kinase-deficient Src acts as a suppressor of the Abl kinase for Cbl phosphorylation. AB - The kinase activity of Abl is known to be regulated by a putative trans-acting inhibitor molecule interacting with the Src homology (SH) 3 domain of Abl. Here we report that the kinase-deficient Src (SrcKD) directly inhibits the tyrosine phosphorylation of Cbl and other cellular proteins by Abl. We found that both the SH2 and SH3 domains of SrcKD are necessary for the suppressor activity toward the Abl kinase phosphorylating Cbl. To suppress the Cbl phosphorylation by Abl, the interaction between the SH3 domain of SrcKD and Cbl is required. This interaction between SrcKD and Cbl is regulated by a closed structure of Cbl. The binding of Abl to the extreme carboxyl-terminal region of Cbl unmasks the binding site of SrcKD to Cbl. This results in a ternary complex that inhibits the Abl-mediated phosphorylation of Cbl by steric hindrance. These results illustrate a mechanism by which the enzymatically inactive Src can exert a biological function in vivo. PMID- 10829063 TI - Ordered intracellular RecA-DNA assemblies: a potential site of in vivo RecA mediated activities. AB - The inducible SOS response increases the ability of bacteria to cope with DNA damage through various DNA repair processes in which the RecA protein plays a central role. Here we present the first study of the morphological aspects that accompany the SOS response in Escherichia coli. We find that induction of the SOS system in wild-type bacteria results in a fast and massive intracellular coaggregation of RecA and DNA into a lateral macroscopic assembly. The coaggregates comprise substantial portions of both the cellular RecA and the DNA complement. The structural features of the coaggregates and their relation to in vitro RecA-DNA networks, as well as morphological studies of strains carrying RecA mutants, are all consistent with the possibility that the intracellular assemblies represent a functional entity in which RecA-mediated DNA repair and protection activities occur. PMID- 10829064 TI - Identification of a PDZ-domain-containing protein that interacts with the scavenger receptor class B type I. AB - The scavenger receptor class B type I (SR-BI) mediates the selective uptake of cholesteryl esters from high-density lipoprotein (HDL) and cholesterol secretion into bile in the liver. In this study, we identified an SR-BI-associated protein from rat liver membrane extracts by using an affinity chromatography technique. This protein of 523 amino acids contains four PDZ domains and associates with the C terminus of SR-BI by using its N-terminal first PDZ domain. Therefore, we denoted this protein as CLAMP (C-terminal linking and modulating protein). CLAMP was located mostly in the sinusoidal membranes, whereas SR-BI was detected in both sinusoidal and canalicular membranes. After the solubilization of the liver membranes with Triton X-100, SR-BI was immunoprecipitated with anti-CLAMP monoclonal antibody, suggesting the association of these proteins in vivo. By coexpressing SR-BI with CLAMP in Chinese hamster ovary cells, we observed (i) the increase in the expression level of SR-BI, (ii) the reduction in the deacylation rate of the cholesteryl esters taken up from HDL, and (iii) the change in the intracellular distribution of fluorescent lipid 1,1'-dioctadecyl-3,3, 3',3' tetramethylindocarbocyanine percholate taken up from HDL. Taken together, these data suggest that CLAMP, a four-PDZ-domain-containing protein, is associated with SR-BI in the liver sinusoidal plasma membranes and may modulate the intracellular transport and metabolism of cholesteryl esters taken up from HDL. PMID- 10829065 TI - Expression profiling reveals distinct sets of genes altered during induction and regression of cardiac hypertrophy. AB - Although cardiac hypertrophy has been the subject of intensive investigation, regression of hypertrophy has been significantly less studied, precluding large scale analysis of the relationship between these processes. In the present study, using pharmacological models of cardiac hypertrophy in mice, expression profiling was performed with fragments of more than 4,000 genes to characterize and contrast expression changes during induction and regression of hypertrophy. Administration of angiotensin II and isoproterenol by osmotic minipump produced increases in heart weight (15 and 45%, respectively) that returned to preinduction size after drug withdrawal. From multiple expression analyses of left ventricular RNA isolated at daily time-points during cardiac hypertrophy and regression, we identified sets of genes whose expression was altered at specific stages of this process. While confirming the participation of 25 genes or pathways previously shown to be altered by hypertrophy, a larger set of 30 genes was identified whose expression had not previously been associated with cardiac hypertrophy or regression. Of the 55 genes that showed reproducible changes during the time course of induction and regression, 32 genes were altered only during induction, and 8 were altered only during regression. This study identified both known and novel genes whose expression is affected at different stages of cardiac hypertrophy and regression and demonstrates that cardiac remodeling during regression utilizes a set of genes that are distinct from those used during induction of hypertrophy. PMID- 10829066 TI - Suppressor of cytokine signaling-3 preferentially binds to the SHP-2-binding site on the shared cytokine receptor subunit gp130. AB - Suppressor of cytokine signaling-3 (SOCS-3) is one member of a family of intracellular inhibitors of signaling pathways initiated by cytokines that use, among others, the common receptor subunit gp130. The SH2 domain of SOCS-3 has been shown to be essential for this inhibitory activity, and we have used a quantitative binding analysis of SOCS-3 to synthetic phosphopeptides to map the potential sites of interaction of SOCS-3 with different components of the gp130 signaling pathway. The only high-affinity ligand found corresponded to the region of gp130 centered around phosphotyrosine-757 (pY757), previously shown to be a docking site for the tyrosine phosphatase SHP-2. By contrast, phosphopeptides corresponding to other regions within gp130, Janus kinase, or signal transducer and activator of transcription proteins bound to SOCS-3 with weak or undetectable affinity. The significance of pY757 in gp130 as a biologically relevant SOCS-3 docking site was investigated by using transfected 293T fibroblasts. Although SOCS-3 inhibited signaling in cells transfected with a chimeric receptor containing the wild-type gp130 intracellular domain, inhibition was considerably impaired for a receptor carrying a Y-->F point mutation at residue 757. Taken together, these data suggest that the mechanism by which SOCS-3 inhibits the gp130 signaling pathway depends on recruitment to the phosphorylated gp130 receptor, and that some of the negative regulatory roles previously attributed to the phosphatase SHP-2 might in fact be caused by the action of SOCS-3. PMID- 10829067 TI - A brain-enriched polypyrimidine tract-binding protein antagonizes the ability of Nova to regulate neuron-specific alternative splicing. AB - The Nova paraneoplastic antigens are neuron-specific RNA binding proteins that participate in the control of alternative splicing. We have used the yeast two hybrid system to isolate Nova interacting proteins and identify an RNA binding protein that is closely related to the polypyrimidine tract-binding protein (PTB). The expression of this protein, brPTB, is enriched in the brain, where it is expressed in glia and neurons. brPTB interacts with Nova proteins in cell lines and colocalizes with Nova within neuronal nuclei. We previously found that Nova binds to a pyrimidine-rich RNA element present upstream of an alternatively spliced exon, E3A, in glycine receptor alpha2 (GlyRalpha2) pre-mRNA, and this binding is implicated in Nova-dependent regulation of splicing. Cotransfection assays with a GlyRalpha2 minigene demonstrate that brPTB antagonizes the action of Nova to increase utilization of GlyRalpha2 E3A. brPTB binds to a 90-nt GlyRalpha2 RNA adjacent to the Nova binding site, but with an affinity that is more than 10-fold lower than Nova. When a putative binding site for brPTB on the GlyRalpha2 RNA is mutated, binding is abolished and the inhibitory effect on Nova dependent exon selection disappears. These results suggest that brPTB is a tissue restricted RNA binding protein that interacts with and inhibits the ability of Nova to activate exon selection in neurons. PMID- 10829068 TI - Inhibition of huntingtin fibrillogenesis by specific antibodies and small molecules: implications for Huntington's disease therapy. AB - The accumulation of insoluble protein aggregates in intra and perinuclear inclusions is a hallmark of Huntington's disease (HD) and related glutamine repeat disorders. A central question is whether protein aggregation plays a direct role in the pathogenesis of these neurodegenerative diseases. Here we show by using a filter retardation assay that the mAb 1C2, which specifically recognizes the elongated polyglutamine (polyQ) stretch in huntingtin, and the chemical compounds Congo red, thioflavine S, chrysamine G, and Direct fast yellow inhibit HD exon 1 protein aggregation in a dose-dependent manner. On the other hand, potential inhibitors of amyloid-beta formation such as thioflavine T, gossypol, melatonin, and rifampicin had little or no inhibitory effect on huntingtin aggregation in vitro. The results obtained by the filtration assay were confirmed by electron microscopy, SDS/PAGE, and MS. Furthermore, cell culture studies revealed that the Congo red dye at micromolar concentrations reduced the extent of HD exon 1 aggregation in transiently transfected COS cells. Together, these findings contribute to a better understanding of the mechanism of huntingtin fibrillogenesis in vitro and provide the basis for the development of new huntingtin aggregation inhibitors that may be effective in treating HD. PMID- 10829069 TI - Heterologous reconstitution in yeast of the polyunsaturated fatty acid biosynthetic pathway. AB - A Caenorhabditis elegans ORF encoding the presumptive condensing enzyme activity of a fatty acid elongase has been characterized functionally by heterologous expression in yeast. This ORF (F56H11. 4) shows low similarity to Saccharomyces cerevisiae genes involved in fatty acid elongation. The substrate specificity of the C. elegans enzyme indicated a preference for Delta(6)-desaturated C18 polyunsaturated fatty acids. Coexpression of this activity with fatty acid desaturases required for the synthesis of C20 polyunsaturated fatty acids resulted in the accumulation of arachidonic acid from linoleic acid and eicosapentaenoic acid from alpha-linolenic acid. These results demonstrate the reconstitution of the n-3 and n-6 polyunsaturated fatty acid biosynthetic pathways. The C. elegans ORF is likely to interact with endogenous components of a yeast elongation system, with the heterologous nematode condensing enzyme F56H11.4 causing a redirection of enzymatic activity toward polyunsaturated C18 fatty acid substrates. PMID- 10829071 TI - An acyltransferase catalyzing the formation of diacylglucose is a serine carboxypeptidase-like protein. AB - 1-O-beta-acyl acetals serve as activated donors in group transfer reactions involved in plant natural product biosynthesis and hormone metabolism. However, the acyltransferases that mediate transacylation from 1-O-beta-acyl acetals have not been identified. We report the identification of a cDNA encoding a 1-O-beta acylglucose-dependent acyltransferase functioning in glucose polyester biosynthesis by Lycopersicon pennellii. The acyltransferase cDNA encodes a serine carboxypeptidase-like protein, with a conserved Ser-His-Asp catalytic triad. Expression of the acyltransferase cDNA in Saccharomyces cerevisiae conferred the ability to disproportionate 1-O-beta-acylglucose to diacylglucose. The disproportionation reaction is regiospecific, catalyzing the conversion of two equivalents of 1-O-beta-acylglucose to 1, 2-di-O-acylglucose and glucose. Diisopropyl fluorophosphate, a transition-state analog inhibitor of serine carboxypeptidases, inhibited acyltransferase activity and covalently labeled the purified acyltransferase, suggesting the involvement of an active serine in the mechanism of the transacylation. The acyltransferase exhibits no carboxypeptidase activity; conversely, the serine carboxypeptidases we have tested show no ability to transacylate using 1-O-acyl-beta-glucoses. This acyltransferase may represent one member of a broader class of enzymes recruited from proteases that have adapted a common catalytic mechanism of catabolism and modified it to accommodate a wide range of group transfer reactions used in biosynthetic reactions of secondary metabolism. The abundance of serine carboxypeptidase-like proteins in plants suggests that this motif has been used widely for metabolic functions. PMID- 10829070 TI - A sensitized genetic system for the analysis of murine B lymphocyte signal transduction pathways dependent on Bruton's tyrosine kinase. AB - Modifier screens have been powerful genetic tools to define signaling pathways in lower organisms. The identification of modifier loci in mice has begun to allow a similar dissection of mammalian signaling pathways. Transgenic mice (Btk(lo)) expressing 25% of endogenous levels of Bruton's tyrosine kinase (Btk) have B cell functional responses between those of wild-type and Btk(-/-) mice. We asked whether reduced dosage or complete deficiency of genes previously implicated as Btk regulators would modify the Btk(lo) phenotype. We used two independent assays of Btk-dependent B cell function. Proliferative response to B cell antigen receptor cross-linking in vitro was chosen as an example of a relatively simple, well-defined signaling system. In vivo response to type II T-independent antigens (TI-II) measures complex interactions among multiple cell types over time and may identify additional Btk pathways. All modifiers identified differentially affected these two assays, indicating that Btk mediates these processes via distinct mechanisms. Loss of Lyn, PTEN (phosphatase and tensin homolog), or SH2 containing inositol phosphatase suppressed the Btk(lo) phenotype in vitro but not in vivo, whereas CD19 and the p85alpha form of phosphoinositide 3-kinase behaved as Btk(lo) enhancers in vivo but not in vitro. Effects of Lyn, PTEN, or p85alpha haploinsufficiency were observed. Haploinsufficiency or complete deficiency of protein kinase C beta, Fyn, CD22, Galphaq, or Galpha11 had no detectable effect on the function of Btk(lo) B cells. A transgenic system creating a reduction in dosage of Btk can therefore be used to identify modifier loci that affect B cell responses and quantitatively rank their contribution to Btk-mediated processes. PMID- 10829072 TI - Induction of apoptosis in human papillomaviruspositive cancer cells by peptide aptamers targeting the viral E6 oncoprotein. AB - Certain types of human papillomaviruses (HPVs) are closely linked to the development of human cancers. Herein, it is shown that intracellular targeting of the HPV16 E6 oncoprotein by E6-binding peptide aptamers resulted in the apoptotic elimination of HPV16-positive cancer cells, whereas HPV-negative cells were not affected. These results provide direct experimental evidence that the HPV E6 oncoprotein has antiapoptotic activity in HPV-positive tumor cells that is required for their survival. The E6-targeting molecules identified herein have implications for the development of therapeutic strategies for the treatment of HPV-associated dysplasias and cancers. PMID- 10829073 TI - Parathyroid hormone-related protein induces spontaneous osteoclast formation via a paracrine cascade. AB - Experiments in vivo have established that tooth eruption fails in the absence of parathyroid hormone (PTH)-related protein (PTHrP) action in the microenvironment of the tooth because of the failure of osteoclastic bone resorption on the coronal tooth surface to form an eruption pathway. To elucidate the effects of PTHrP on osteoclast regulation in this environment, we established primary cultures of epithelial stellate reticulum cells and mesenchymal dental follicle (DF) cells surrounding the teeth. When cocultured, these cells are fully capable of supporting the formation of functional osteoclasts in the absence of added splenic osteoclast precursors, osteoblasts, or vitamin D/PTH/PTHrP. Neutralizing the effects of PTHrP resulted in a decrease in the number of osteoclasts formed, suggesting that stellate reticulum-derived PTHrP drives osteoclast formation. DF cells were found to express functional PTH/PTHrP type I receptors, and conditioned media collected from PTHrP-treated DF cells were able to induce bone resorption in the fetal-rat long-bone assay. PTHrP treatment also induced an increase in osteoclast differentiation factor expression and a concomitant decrease in osteoclastogenesis inhibitory factor expression in DF cells. The addition of osteoclastogenesis inhibitory factor resulted in a decrease in the number of osteoclasts formed in the cocultures, suggesting that osteoclast formation is mediated by osteoclast differentiation factor. Thus, PTHrP seems to regulate osteoclast formation via mediation of the DF, in a manner analogous to the osteoblast-mediated process in the peripheral skeleton. The primary coculture system of dental crypt cells also offers a system for the study of osteoclast formation and regulation. PMID- 10829074 TI - The complete optical spectrum of liquid water measured by inelastic x-ray scattering. AB - Interaction of light with matter is of paramount importance in nature. The most fundamental property of a material in relation to light is its oscillator strength distribution, i.e., how strongly it absorbs light as a function of wavelength. Once the oscillator strength distribution is determined precisely for a wide enough energy range, the optical constants such as absorbance and reflectance as well as a number of other properties of the material, some of which are seemingly unrelated to photoabsorption, can be deduced. Most important of all is the fact that the interaction of matter with fast charged particles can be described by its complete optical spectra [Inokuti, M. (1986) Photochem. Photobiol. 44, 279-285]. Despite their importance, however, the complete optical spectra of volatile liquids including water have never been obtained accurately because of experimental difficulties inherent in vacuum UV spectroscopy. Inelastic x-ray scattering spectroscopy can provide quantitative data equivalent to those from vacuum UV absorption spectra. Herein, we show the complete optical spectrum of liquid water determined by making use of intense monochromatic x-rays supplied by the wiggler line X21 of the National Synchrotron Light Source. PMID- 10829075 TI - Phospholipid:diacylglycerol acyltransferase: an enzyme that catalyzes the acyl CoA-independent formation of triacylglycerol in yeast and plants. AB - Triacylglycerol (TAG) is known to be synthesized in a reaction that uses acyl-CoA as acyl donor and diacylglycerol (DAG) as acceptor, and which is catalyzed by the enzyme acyl-CoA:diacylglycerol acyltransferase. We have found that some plants and yeast also have an acyl-CoA-independent mechanism for TAG synthesis, which uses phospholipids as acyl donors and DAG as acceptor. This reaction is catalyzed by an enzyme that we call phospholipid:diacylglycerol acyltransferase, or PDAT. PDAT was characterized in microsomal preparations from three different oil seeds: sunflower, castor bean, and Crepis palaestina. We found that the specificity of the enzyme for the acyl group in the phospholipid varies between these species. Thus, C. palaestina PDAT preferentially incorporates vernoloyl groups into TAG, whereas PDAT from castor bean incorporates both ricinoleoyl and vernoloyl groups. We further found that PDAT activity also is present in yeast microsomes. The substrate specificity of this PDAT depends on the head group of the acyl donor, the acyl group transferred, and the acyl chains of the acceptor DAG. The gene encoding the enzyme was identified. The encoded PDAT protein is related to lecithin:cholesterol acyltransferase, which catalyzes the acyl-CoA-independent synthesis of cholesterol esters. However, budding yeast PDAT and its relatives in fission yeast and Arabidopsis form a distinct branch within this protein superfamily, indicating that a separate PDAT enzyme arose at an early point in evolution. PMID- 10829076 TI - Prothrombotic phenotype of protein Z deficiency. AB - Protein Z (PZ) is a vitamin K-dependent plasma protein whose function has been uncertain. The structure of PZ is very similar to that of the coagulation-related factors VII, IX, and X and PC, but PZ differs from these other proteins in that it is not the zymogen of a serine protease. We have shown recently that PZ forms a calcium ion-dependent complex with activated factor X at phospholipid surfaces and that this interaction leads to the inhibition of activated factor X activity through, in part, the action of a previously unidentified plasma protein named PZ dependent protease inhibitor. Herein, we report that the presence of PZ dampens the coagulation response in human plasma and that concomitant PZ deficiency dramatically increases the severity of the prothrombotic phenotype of factor V(Leiden) mice. The results indicate that PZ plays a physiologically important role in the regulation of coagulation. PMID- 10829077 TI - The SOS response regulates adaptive mutation. AB - Upon starvation some Escherichia coli cells undergo a transient, genome-wide hypermutation (called adaptive mutation) that is recombination-dependent and appears to be a response to a stressful environment. Adaptive mutation may reflect an inducible mechanism that generates genetic variability in times of stress. Previously, however, the regulatory components and signal transduction pathways controlling adaptive mutation were unknown. Here we show that adaptive mutation is regulated by the SOS response, a complex, graded response to DNA damage that includes induction of gene products blocking cell division and promoting mutation, recombination, and DNA repair. We find that SOS-induced levels of proteins other than RecA are needed for adaptive mutation. We report a requirement of RecF for efficient adaptive mutation and provide evidence that the role of RecF in mutation is to allow SOS induction. We also report the discovery of an SOS-controlled inhibitor of adaptive mutation, PsiB. These results indicate that adaptive mutation is a tightly regulated response, controlled both positively and negatively by the SOS system. PMID- 10829078 TI - The t(1;12)(q21;p13) translocation of human acute myeloblastic leukemia results in a TEL-ARNT fusion. AB - The TEL/ETV6 gene is located at 12p13 and encodes a member of the ETS family of transcription factors. Translocated ETS leukemia (TEL) is frequently involved in chromosomal translocations in human malignancies, usually resulting in the expression of fusion proteins between the amino-terminal part of TEL and either unrelated transcription factors or protein tyrosine kinases. We have characterized a t(1;12)(q21;p13) translocation in an acute myeloblastic leukemia (AML-M2). At the protein level, the untranslocated TEL copy and, as a result of the t(1;12) translocation, a fusion protein between TEL and essentially all of aryl hydrocarbon receptor nuclear translocator (ARNT) are expressed. The involvement of ARNT in human leukemogenesis has not been previously described. The ARNT protein belongs to a subfamily of the "basic region helix-loop-helix" (bHLH) protein that shares an additional region of similarity called the PAS (Per, ARNT, SIM) domain. ARNT is the central partner of several heterodimeric transcription factors, including those containing the aryl hydrocarbon (dioxin) receptor (AhR) and the hypoxia-inducible factor 1alpha (HIF1alpha). Our results show that the TEL-ARNT fusion protein is the crucial product of the translocation and suggest that interference with the activity of AhR or HIF1alpha can contribute to leukemogenesis. PMID- 10829079 TI - One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. AB - We have developed a simple and highly efficient method to disrupt chromosomal genes in Escherichia coli in which PCR primers provide the homology to the targeted gene(s). In this procedure, recombination requires the phage lambda Red recombinase, which is synthesized under the control of an inducible promoter on an easily curable, low copy number plasmid. To demonstrate the utility of this approach, we generated PCR products by using primers with 36- to 50-nt extensions that are homologous to regions adjacent to the gene to be inactivated and template plasmids carrying antibiotic resistance genes that are flanked by FRT (FLP recognition target) sites. By using the respective PCR products, we made 13 different disruptions of chromosomal genes. Mutants of the arcB, cyaA, lacZYA, ompR-envZ, phnR, pstB, pstCA, pstS, pstSCAB-phoU, recA, and torSTRCAD genes or operons were isolated as antibiotic-resistant colonies after the introduction into bacteria carrying a Red expression plasmid of synthetic (PCR-generated) DNA. The resistance genes were then eliminated by using a helper plasmid encoding the FLP recombinase which is also easily curable. This procedure should be widely useful, especially in genome analysis of E. coli and other bacteria because the procedure can be done in wild-type cells. PMID- 10829080 TI - Severe deficiencies in dopamine signaling in presymptomatic Huntington's disease mice. AB - In Huntington's disease (HD), mutation of huntingtin causes selective neurodegeneration of dopaminoceptive striatal medium spiny neurons. Transgenic HD model mice that express a portion of the disease-causing form of human huntingtin develop a behavioral phenotype that suggests dysfunction of dopaminergic neurotransmission. Here we show that presymtomatic mice have severe deficiencies in dopamine signaling in the striatum. These include selective reductions in total levels of dopamine- and cAMP-regulated phosphoprotein, M(r) 32 kDA (DARPP 32) and other dopamine-regulated phosphoprotein markers of medium spiny neurons. HD mice also show defects in dopamine-regulated ion channels and in the D(1) dopamine/DARPP-32 signaling cascade. These presymptomatic defects may contribute to HD pathology. PMID- 10829081 TI - Copy-choice recombination by reverse transcriptases: reshuffling of genetic markers mediated by RNA chaperones. AB - Copy-choice recombination efficiently reshuffles genetic markers in retroviruses. In vivo, the folding of the genomic RNA is controlled by the nucleocapsid protein (NC). We show that binding of NC onto the acceptor RNA molecule is sufficient to enhance recombination, providing evidence for a mechanism where the structure of the acceptor template determines the template switch. NC as well as another RNA chaperone (StpA) converts recombination into a widespread process no longer restricted to rare hot spots, an effect maximized when both the NC and the reverse transcriptase come from HIV-1. These data suggest that RNA chaperones confer a higher genetic flexibility to retroviruses. PMID- 10829082 TI - B cell receptor expression level determines the fate of developing B lymphocytes: receptor editing versus selection. AB - During B lymphocyte development, antibody genes are assembled by DNA recombination. Successful cell surface expression of IgM promotes developmental progression. However, when antigen receptors bind autoantigen, development is blocked and ongoing antibody gene recombination occurs, which often alters antibody specificity in a process called receptor editing. We demonstrate here a significant role of developmental block and receptor editing in B cell receptor quality control. During development a functional, non-self-reactive receptor undergoes receptor editing if its expression is below a certain threshold. Doubling the receptor gene dose promotes development in the absence of autoantigen, but allows editing when autoantigen is present. Thus, both underexpressed and harmful B cell receptors can undergo correction by receptor editing. PMID- 10829083 TI - Analysis of telomerase catalytic subunit mutants in vivo and in vitro in Schizosaccharomycespombe. AB - The chromosome end-replicating enzyme telomerase is composed of a template containing RNA subunit, a reverse transcriptase (TERT), and additional proteins. The importance of conserved amino acid residues in Trt1p, the TERT of Schizosaccharomyces pombe, was tested. Mutation to alanine of the proposed catalytic aspartates in reverse transcriptase motifs A and C and of conserved amino acids in motifs 1 and B' resulted in defective growth, progressive loss of telomeric DNA, and loss of detectable telomerase enzymatic activity in vitro. Mutation of the phenylalanine (F) in the conserved FYxTE of telomerase-specific motif T had no phenotype in vivo or in vitro whereas mutation of a conserved amino acid in RT motif 2 had an intermediate effect. In addition to identifying single amino acids of TERT required for telomere maintenance in the fission yeast, this work provides useful tools for S. pombe telomerase research: a functional epitope-tagged version of Trt1p that allows detection of the protein even in crude cellular extracts, and a convenient and robust in vitro enzymatic activity assay based on immunopurification of telomerase. PMID- 10829084 TI - The simulated evolution of biochemical guilds: reconciling Gaia theory and natural selection. AB - Gaia theory, which states that organisms both affect and regulate their environment, poses an interesting problem to Neo-Darwinian evolutionary biologists and provides an exciting set of phenomena for artificial-life investigation. The key challenge is to explain the emergence of biotic communities that are capable, via their implicit coordination, of regulating large-scale biogeochemical factors such as the temperature and chemical composition of the biosphere, but to assume no evolutionary mechanisms beyond contemporary natural selection. Along with providing an introduction to Gaia theory, this article presents simulations of Gaian emergence based on an artificial-life model involving genetic algorithms and guilds of simple metabolizing agents. In these simulations, resource competition leads to guild diversity; the ensemble of guilds then manifests life-sustaining nutrient recycling and exerts distributed control over environmental nutrient ratios. These results illustrate that standard individual-based natural selection is sufficient to explain Gaian self-organization, and they help clarify the relationships between two key metrics of Gaian activity: recycling and regulation. PMID- 10829085 TI - An emergence of coordinated communication in populations of agents. AB - The purpose of this article is to demonstrate that coordinated communication spontaneously emerges in a population composed of agents that are capable of specific cognitive activities. Internal states of agents are characterized by meaning vectors. Simple neural networks composed of one layer of hidden neurons perform cognitive activities of agents. An elementary communication act consists of the following: (a) two agents are selected, where one of them is declared the speaker and the other the listener; (b) the speaker codes a selected meaning vector onto a sequence of symbols and sends it to the listener as a message; and finally, (c) the listener decodes this message into a meaning vector and adapts his or her neural network such that the differences between speaker and listener meaning vectors are decreased. A Darwinian evolution enlarged by ideas from the Baldwin effect and Dawkins' memes is simulated by a simple version of an evolutionary algorithm without crossover. The agent fitness is determined by success of the mutual pairwise communications. It is demonstrated that agents in the course of evolution gradually do a better job of decoding received messages (they are closer to meaning vectors of speakers) and all agents gradually start to use the same vocabulary for the common communication. Moreover, if agent meaning vectors contain regularities, then these regularities are manifested also in messages created by agent speakers, that is, similar parts of meaning vectors are coded by similar symbol substrings. This observation is considered a manifestation of the emergence of a grammar system in the common coordinated communication. PMID- 10829086 TI - A new structurally dissolvable self-reproducing loop evolving in a simple cellular automata space. AB - We constructed a simple evolutionary system, "evoloop," on a deterministic nine state five-neighbor cellular automata (CA) space by improving the structurally dissolvable self-reproducing loop we had previously contrived [14] after Langton's self-reproducing loop [7]. The principal role of this improvement is to enhance the adaptability (a degree of the variety of situations in which structures in the CA space can operate regularly) of the self-reproductive mechanism of loops. The experiment with evoloop met with the intriguing result that, though no mechanism was explicitly provided to promote evolution, the loops varied through direct interaction of their phenotypes, smaller individuals were naturally selected thanks to their quicker self-reproductive ability, and the whole population gradually evolved toward the smallest ones. This result gives a unique example of evolution of self-replicators where genotypical variation is caused by precedent phenotypical variation. Such interrelation of genotype and phenotype would be one of the important factors driving the evolutionary process of primitive life forms that might have actually occurred in ancient times. PMID- 10829087 TI - An approach to biological computation: unicellular core-memory creatures evolved using genetic algorithms. AB - A novel machine language genetic programming system that uses one-dimensional core memories is proposed and simulated. The core is compared to a biochemical reaction space, and in imitation of biological molecules, four types of data words (Membrane, Pure data, Operator, and Instruction) are prepared in the core. A program is represented by a sequence of Instructions. During execution of the core, Instructions are transcribed into corresponding Operators, and Operators modify, create, or transfer Pure data. The core is hierarchically partitioned into sections by the Membrane data, and the data transfer between sections by special channel Operators constitutes a tree data-flow structure among sections in the core. In the experiment, genetic algorithms are used to modify program information. A simple machine learning problem is prepared for the environment data set of the creatures (programs), and the fitness value of a creature is calculated from the Pure data excreted by the creature. Breeding of programs that can output the predefined answer is successfully carried out. Several future plans to extend this system are also discussed. PMID- 10829088 TI - Role of sympathetic nervous system and neuropeptides in obesity hypertension. AB - Obesity is the most common cause of human essential hypertension in most industrialized countries. Although the precise mechanisms of obesity hypertension are not fully understood, considerable evidence suggests that excess renal sodium reabsorption and a hypertensive shift of pressure natriuresis play a major role. Sympathetic activation appears to mediate at least part of the obesity-induced sodium retention and hypertension since adrenergic blockade or renal denervation markedly attenuates these changes. Recent observations suggest that leptin and its multiple interactions with neuropeptides in the hypothalamus may link excess weight gain with increased sympathetic activity. Leptin is produced mainly in adipocytes and is believed to regulate energy balance by acting on the hypothalamus to reduce food intake and to increase energy expenditure via sympathetic activation. Short-term administration of leptin into the cerebral ventricles increases renal sympathetic activity, and long-term leptin infusion at rates that mimic plasma concentrations found in obesity raises arterial pressure and heart rate via adrenergic activation in non-obese rodents. Transgenic mice overexpressing leptin also develop hypertension. Acute studies suggest that the renal sympathetic effects of leptin may depend on interactions with other neurochemical pathways in the hypothalamus, including the melanocortin-4 receptor (MC4-R). However, the role of this pathway in mediating the long-term effects of leptin on blood pressure is unclear. Also, it is uncertain whether there is resistance to the chronic renal sympathetic and blood pressure effects of leptin in obese subjects. In addition, leptin also has other cardiovascular and renal actions, such as stimulation of nitric oxide formation and improvement of insulin sensitivity, which may tend to reduce blood pressure in some conditions. Although the role of these mechanisms in human obesity has not been elucidated, this remains a fruitful area for further investigation, especially in view of the current "epidemic" of obesity in most industrialized countries. PMID- 10829089 TI - Angiotensin II-mediated vascular smooth muscle cell growth signaling. AB - The mechanism by which Ang II stimulates the growth of vascular smooth muscle cells was investigated by measuring the phosphorylation of mitogen-activated protein kinases ERK 1 and ERK 2. Ca2+ ionophore was found to have effects practically analogous to Ang II. We found that the signaling pathway involves the activation of epidermal growth factor receptor (EGFR) kinase, activation of the adaptor proteins Shc and Grb2, and the small G-protein Ras. Although the mechanism of AT1- (or Ca2+)-induced activation of EGFR is not yet clear, we have found that calcium-dependent protein kinase CAKss/PYK2 and c-Src are involved in this process. These studies indicate a transactivation mechanism that utilizes EGFR as a bridge between a Gq-coupled receptor and activation of phosphotyrosine generation. PMID- 10829090 TI - Oxytocin is a cardiovascular hormone. AB - Oxytocin (OT), a nonapeptide, was the first hormone to have its biological activities established and chemical structure determined. It was believed that OT is released from hypothalamic nerve terminals of the posterior hypophysis into the circulation where it stimulates uterine contractions during parturition, and milk ejection during lactation. However, equivalent concentrations of OT were found in the male hypophysis, and similar stimuli of OT release were determined for both sexes, suggesting other physiological functions. Indeed, recent studies indicate that OT is involved in cognition, tolerance, adaptation and complex sexual and maternal behaviour, as well as in the regulation of cardiovascular functions. It has long been known that OT induces natriuresis and causes a fall in mean arterial pressure, both after acute and chronic treatment, but the mechanism was not clear. The discovery of the natriuretic family shed new light on this matter. Atrial natriuretic peptide (ANP), a potent natriuretic and vasorelaxant hormone, originally isolated from rat atria, has been found at other sites, including the brain. Blood volume expansion causes ANP release that is believed to be important in the induction of natriuresis and diuresis, which in turn act to reduce the increase in blood volume. Neurohypophysectomy totally abolishes the ANP response to volume expansion. This indicates that one of the major hypophyseal peptides is responsible for ANP release. The role of ANP in OT induced natriuresis was evaluated, and we hypothesized that the cardio-renal effects of OT are mediated by the release of ANP from the heart. To support this hypothesis, we have demonstrated the presence and synthesis of OT receptors in all heart compartments and the vasculature. The functionality of these receptors has been established by the ability of OT to induce ANP release from perfused heart or atrial slices. Furthermore, we have shown that the heart and large vessels like the aorta and vena cava are sites of OT synthesis. Therefore, locally produced OT may have important regulatory functions within the heart and vascular beds. Such functions may include slowing down of the heart or the regulation of local vascular tone. PMID- 10829091 TI - Effects of exercise training on autonomic and myocardial dysfunction in streptozotocin-diabetic rats. AB - Several investigators have demonstrated that diabetes is associated with autonomic and myocardial dysfunction. Exercise training is an efficient non pharmacological treatment for cardiac and metabolic diseases. The aim of the present study was to investigate the effects of exercise training on hemodynamic and autonomic diabetic dysfunction. After 1 week of diabetes induction (streptozotocin, 50 mg/kg, iv), male Wistar rats (222 +/- 5 g, N = 18) were submitted to exercise training for 10 weeks on a treadmill. Arterial pressure signals were obtained and processed with a data acquisition system. Autonomic function and intrinsic heart rate were studied by injecting methylatropine and propranolol. Left ventricular function was assessed in hearts perfused in vitro by the Langendorff technique. Diabetes (D) bradycardia and hypotension (D: 279 +/ 9 bpm and 91 +/- 4 mmHg vs 315 +/- 11 bpm and 111 +/- 4 mmHg in controls, C) were attenuated by training (TD: 305 +/- 7 bpm and 100 +/- 4 mmHg). Vagal tonus was decreased in the diabetic groups and sympathetic tonus was similar in all animals. Intrinsic heart rate was lower in D (284 +/- 11 bpm) compared to C and TD (390 +/- 8 and 342 +/- 14 bpm, respectively). Peak systolic pressure developed at different pressures was similar for all groups, but +dP/dt max was decreased and -dP/dt max was increased in D. In conclusion, exercise training reversed hypotension and bradycardia and improved myocardial function in diabetic rats. These changes represent an adaptive response to the demands of training, supporting a positive role of physical activity in the management of diabetes. PMID- 10829092 TI - The physiological role of AT1 receptors in the ventrolateral medulla. AB - Neurons in the rostral and caudal parts of the ventrolateral medulla (VLM) play a pivotal role in the regulation of sympathetic vasomotor activity and blood pressure. Studies in several species, including humans, have shown that these regions contain a high density of AT1 receptors specifically associated with neurons that regulate the sympathetic vasomotor outflow, or the secretion of vasopressin from the hypothalamus. It is well established that specific activation of AT1 receptors by application of exogenous angiotensin II in the rostral and caudal VLM excites sympathoexcitatory and sympathoinhibitory neurons, respectively, but the physiological role of these receptors in the normal synaptic regulation of VLM neurons is not known. In this paper we review studies which have defined the effects of specific activation or blockade of these receptors on cardiovascular function, and discuss what these findings tell us with regard to the physiological role of AT1 receptors in the VLM in the tonic and phasic regulation of sympathetic vasomotor activity and blood pressure. PMID- 10829093 TI - Oxidative stress may explain how hypertension is maintained by normal levels of angiotensin II. AB - It is well known that essential hypertension evolves in most patients with "near normal" levels of plasma renin activity. However, these levels appear to be responsible for the high levels of arterial pressure because they are normalized by the administration of angiotensin II converting inhibitors or angiotensin receptor antagonist. In experimental animals, hypertension can be induced by the continuous intravenous infusion of small doses of angiotensin II that are not sufficient to evoke an immediate pressor response. However, this condition resembles the characteristics of essential hypertension because the high levels of blood pressure exist with normal plasma levels of angiotensin II. It is suggested that small amounts of angiotensin whose plasma levels are inappropriate for the existing size of extracellular volume stimulate oxidative stress which binds nitric oxide forming peroxynitrite. The latter compound oxidizes arachidonic acid producing isoprostaglandin F2alpha (an isoprostane) which is characterized by a strong antinatriuretic vasoconstrictor renal effect. In this chain of reactions the vasoconstrictor effects derived from oxygen quenching of nitric oxide and increased isoprostane synthesis could explain how hypertension is maintained with normal plasma levels of renin. PMID- 10829094 TI - Stimulatory effect of dexamethasone on angiotensin-converting enzyme in neonatal rat cardiac myocytes. AB - Angiotensin-converting enzyme (ACE) plays a central role in cardiac remodeling associated with pathological conditions such as myocardial infarction. The existence of different cell types in the heart expressing components of the renin angiotensin system makes it difficult to evaluate their relative role under physiological and pathological conditions. Since myocytes are the predominant cellular constituent of the heart by mass, in the present study we studied the effects of glucocorticoids on ACE activity using well-defined cultures of neonatal rat cardiac myocytes. Under steady-state conditions, ACE activity was present at very low levels, but after dexamethasone treatment ACE activity increased significantly (100 nmol/l after 24 h) in a time-dependent fashion. These results demonstrate the influence of dexamethasone on ACE activity in rat cardiac myocytes. This is consistent with the idea that ACE activation occurs under stress conditions, such as myocardial infarction, in which glucocorticoid levels may increase approximately 50-fold. PMID- 10829095 TI - Towards understanding the kallikrein-kinin system: insights from measurement of kinin peptides. AB - The kallikrein-kinin system is complex, with several bioactive peptides that are formed in many different compartments. Kinin peptides are implicated in many physiological and pathological processes including the regulation of blood pressure and sodium homeostasis, inflammatory processes, and the cardioprotective effects of preconditioning. We established a methodology for the measurement of individual kinin peptides in order to study the function of the kallikrein-kinin system. The levels of kinin peptides in tissues were higher than in blood, confirming the primary tissue localization of the kallikrein-kinin system. Moreover, the separate measurement of bradykinin and kallidin peptides in man demonstrated the differential regulation of the plasma and tissue kallikrein kinin systems, respectively. Kinin peptide levels were increased in the heart of rats with myocardial infarction, in tissues of diabetic and spontaneously hypertensive rats, and in urine of patients with interstitial cystitis, suggesting a role for kinin peptides in the pathogenesis of these conditions. By contrast, blood levels of kallidin, but not bradykinin, peptides were suppressed in patients with severe cardiac failure, suggesting that the activity of the tissue kallikrein-kinin system may be suppressed in this condition. Both angiotensin converting enzyme (ACE) and neutral endopeptidase (NEP) inhibitors increased bradykinin peptide levels. ACE and NEP inhibitors had different effects on kinin peptide levels in blood, urine, and tissues, which may be accounted for by the differential contributions of ACE and NEP to kinin peptide metabolism in the multiple compartments in which kinin peptide generation occurs. Measurement of the levels of individual kinin peptides has given important information about the operation of the kallikrein-kinin system and its role in physiology and disease states. PMID- 10829096 TI - Review of the Y chromosome and hypertension. AB - The Y chromosome from spontaneously hypertensive rats (SHR) has a locus that raises blood pressure 20-25 mmHg. Associated with the SHR Y chromosome effect is a 4-week earlier pubertal rise of testosterone and dependence upon the androgen receptor for the full blood pressure effect. Several indices of enhanced sympathetic nervous system (SNS) activity are also associated with the SHR Y chromosome. Blockade of SNS outflow reduced the blood pressure effect. Salt sensitivity was increased by the Y chromosome as was salt appetite which was SNS dependent. A strong correlation (r = 0. 57, P<0.001) was demonstrable between plasma testosterone and angiotensin II. Coronary collagen increased with blood pressure and the presence of the SHR Y chromosome. A promising candidate gene for the Y effect is the Sry locus (testis determining factor), a transcription factor which may also have other functions. PMID- 10829097 TI - Ischemia and fibrosis: the risk mechanisms of hypertensive heart disease. AB - Mechanisms underlying risk associated with hypertensive heart disease (HHD) and left ventricular hypertrophy (LVH) are discussed in this report and provide a rationale for understanding this very common and important cause of death from hypertension and its complications. Emphasized are impaired coronary hemodynamics, endothelial dysfunction, and ventricular fibrosis from increased collagen deposition intramurally and perivascularly. Each is exacerbated by aging and, perhaps, also by increased dietary salt intake. These functional and structural changes promote further endothelial dysfunction, altered coronary hemodynamics, and diastolic as well as systolic ventricular contractile function in HHD. The clinical endpoints of HHD include angina pectoris (with or without atherosclerosis of the epicardial coronary arteries), myocardial infarction, cardiac failure, lethal dysrhythmias, and sudden death. The major concept to be derived from these alterations is that not all that is clinically recognized as LVH is true myocytic hypertrophy and structural remodeling. Other major co-morbid changes occur that serve to increase cardiovascular risk including impaired coronary hemodynamics, endothelial dysfunction, and ventricular fibrosis. PMID- 10829098 TI - Regulation of the kinin receptors after induction of myocardial infarction: a mini-review. AB - It is well known that the responses to vasoactive kinin peptides are mediated through the activation of two receptors termed bradykinin receptor B1 (B1R) and B2 (B2R). The physiologically prominent B2R subtype has certainly been the subject of more intensive efforts in structure-function studies and physiological investigations. However, the B1R activated by a class of kinin metabolites has emerged as an important subject of investigation within the study of the kallikrein-kinin system (KKS). Its inducible character under stress and tissue injury is therefore a field of major interest. Although the KKS has been associated with cardiovascular regulation since its discovery at the beginning of the last century, less is known about the B1R and B2R regulation in cardiovascular diseases like hypertension, myocardial infarction (MI) and their complications. This mini-review will summarize our findings on B1R and B2R regulation after induction of MI using a rat model. We will develop the hypothesis that differences in the expression of these receptors may be associated with a dual pathway of the KKS in the complex mechanisms of myocardial remodeling. PMID- 10829100 TI - Somatic gene therapy for hypertension. AB - Gene therapy for hypertension is needed for the next generation of antihypertensive drugs. Current drugs, although effective, have poor compliance, are expensive and short-lasting (hours or one day). Gene therapy offers a way to produce long-lasting antihypertensive effects (weeks, months or years). We are currently using two strategies: a) antisense oligodeoxynucleotides (AS-ODN) and b) antisense DNA delivered in viral vectors to inhibit genes associated with vasoconstrictive properties. It is not necessary to know all the genes involved in hypertension, since many years of experience with drugs show which genes need to be controlled. AS-ODN are short, single-stranded DNA that can be injected in naked form or in liposomes. AS-ODN, targeted to angiotensin type 1 receptors (AT1 R), angiotensinogen (AGT), angiotensin converting enzyme, and ss1-adrenergic receptors effectively reduce hypertension in rat models (SHR, 2K-1C) and cold induced hypertension. A single dose is effective up to one month when delivered with liposomes. No side effects or toxic effects have been detected, and repeated injections can be given. For the vector, adeno-associated virus (AAV) is used with a construct to include a CMV promoter, antisense DNA to AGT or AT1-R and a reporter gene. Results in SHR demonstrate reduction and slowing of development of hypertension, with a single dose administration. Left ventricular hypertrophy is also reduced by AAV-AGT-AS treatment. Double transgenic mice (human renin plus human AGT) with high angiotensin II causing high blood pressure, treated with AAV AT1-R-AS, show a normalization of blood pressure for over six months with a single injection of vector. We conclude that ODNs will probably be developed first because they can be treated like drugs for the treatment of hypertension with long-term effects. Viral vector delivery needs more engineering to be certain of its safety, but one day may be used for a very prolonged control of blood pressure. PMID- 10829099 TI - Angiotensin-(1-7) potentiates the coronary vasodilatatory effect of bradykinin in the isolated rat heart. AB - It has been shown that angiotensin-(1-7) (Ang-(1-7)) infusion potentiates the bradykinin (BK)-induced hypotensive response in conscious rats. The present study was conducted to identify Ang-(1-7)-BK interactions in the isolated rat heart perfused according to the Langendorff technique. Hearts were excised and perfused through the aortic stump under a constant flow with Krebs-Ringer solution and the changes in perfusion pressure and heart contractile force were recorded. Bolus injections of BK (2.5, 5, 10 and 20 ng) produced a dose-dependent hypotensive effect. Ang-(1-7) added to the perfusion solution (2 ng/ml) did not change the perfusion pressure or the contractile force but doubled the hypotensive effect of the lower doses of BK. The BK-potentiating Ang-(1-7) activity was blocked by pretreatment with indomethacin (5 mg/kg, ip) or L-NAME (30 mg/kg, ip). The Ang-(1 7) antagonist A-779 (50 ng/ml in Krebs-Ringer) completely blocked the effect of Ang-(1-7) on BK-induced vasodilation. These data suggest that the potentiation of the BK-induced vasodilation by Ang-(1-7) can be attributed to the release of nitric oxide and vasodilator prostaglandins through an Ang-(1-7) receptor mediated mechanism. PMID- 10829101 TI - The future of audiological rehabilitation. AB - The field of audiological rehabilitation in adults faces an array of opportunities. Some of these are technological, as with the advent of fully digital hearing-aids, and some involve clinical practice, such as opportunities for true multidisciplinary working, and for changes in hearing-aid prescription and provision. The development of well-validated questionnaire instruments should facilitate robust research into the effectiveness of clinical interventions in adult audiological rehabilitation, for such evidence is urgently needed if the field is to thrive. PMID- 10829102 TI - The training of otorhinolaryngologists in Asia. AB - Asian nations differ from one another in population, cultural background, ethnic tradition and economic situation. The training of otorhinolaryngologists in Asia is not in any way uniform. A questionnaire survey of 26 leading institutions in 12 countries/regions was carried out. The emphasis of investigation is towards the training programme for specialists in Otorhinolaryngology and continuous medical education (CME). The response rate of the survey was 100 per cent. The duration of training for otorhinolaryngologists ranges from three to seven years and an exit assessment is held in seven countries/regions. The training programmes are modifications of either the British or the American pattern depending on tradition. Documentation of CME is available in eight countries/regions while a defined cycle has been established in only three. Medical/health authorities or professional societies in each country contribute towards its monitoring. Better communication and sharing of experience will improve the training of otorhinolaryngologists in Asia. PMID- 10829103 TI - Myringoplasty: update on onlay pedicle skin flap and temporalis fascia sandwich graft. AB - The technique of onlay sandwich grafting of tympanic membrane perforations fashioned by the use of pedicle flap raised from the posterior deep meatal skin and autologous temporalis fascia is described. The results of a series of 124 ears operated between 1987 and 1999 are the basis of this report. Of these 106 patients had myringoplasty for the first time. The operation was successful in 102 (96.23 per cent) patients. Eighteen patients had revision myringoplasty, with a successful result in 14 (77.78 per cent). None of the patients developed complications usually associated with onlay grafting. A preliminary report of this work was presented at the XVI World Congress of Otolaryngology Head and Neck Surgery at Sydney in 1997. PMID- 10829104 TI - Electronystagmography: how helpful is it? AB - Two hundred and sixty patients tested with electronystagmography (ENG) for evaluation of complaints of disequilibrium were the representative sample for this study. As all new dizzy patients currently receive an ENG, the goal of this project was to determine how much an ENG adds to the diagnostic acumen, and which patients really needed an ENG in order to maximize efficiency and minimize patient cost. Results indicate the suspected diagnosis prior to ENG is assisted by its use only when the cause is thought to be central or is uncertain. The percentage of unknown causes of disequilibrium decreased from 34.2 to 13.8 per cent and the percentage of central causes increased from 13.1 to 23.1 per cent. Electronystagmography does not significantly aid in the diagnosis of peripheral lesions except as confirmation. Specific findings and recommendations are discussed. PMID- 10829105 TI - Microbiology of the middle meatus: a comparison between normal adults and children. AB - Middle meatal samples were obtained from 52 carefully selected healthy adults. In 75 per cent of the test subjects bacterial organisms were cultured. However, growth was often poor and the predominant species suggest a commensal flora: coagulase-negative staphylococci were retrieved from 35 per cent, Corynebacterium sp. from 23 per cent and Staphyloccus aureus from eight per cent of the adults. These data are very different from those previously obtained among children where -even in the absence of obvious ENT pathology--the most frequently cultured organisms were typical sinusitis pathogens: Haemophilus influenzae present in 40 per cent, Moraxella catarrhalis in 34 per cent and Streptococcus pneumoniae in 50 per cent of children. Furthermore, Streptococcus viridans and Neisseria sp., both organisms that might be able to inhibit colonization by some of the pathogens and found commonly among children, are virtually absent in healthy adults. PMID- 10829106 TI - Rhinoplasty in unilateral cleft lip nasal deformity. AB - An operation is described for correction of unilateral cleft lip nasal deformity which has had considerable uniformity of success and is applicable to both mild and severe degrees of deformity. Our proposed repair technique is performed through an external rhinoplasty approach and depends on repositioning of the displaced and deformed cartilages together with the reinforcement of the structural support of the nose by using multiple cartilage grafts. This surgical technique was used in 18 consecutive adult patients with unilateral cleft lip nasal deformity and yielded consistently good long-term functional and cosmetic results. PMID- 10829107 TI - Optimum management of inverted papilloma. AB - Surgery offers the optimum modality of treatment for inverted papilloma although a considerable range of operative approaches have been described. The results are presented in a cohort of 37 cases treated by both endoscopic and combined endoscopic and external approaches with a recurrence rate of eight and 21 per cent respectively. This series is compared with those in the literature and demonstrates that it is extent of disease which primarily determines the choice of surgical approach, with previous treatment, individual patient factors and surgical expertise as secondary determinants. PMID- 10829109 TI - A national survey of disinfection techniques for flexible nasendoscopes in UK ENT out-patient departments. AB - Flexible fibre-optic nasendoscopes have become a ubiquitous ENT out-patient tool for the inspection of the nasopharynx, larynx and hypopharynx. Disinfection of the instrument between patient use is important to prevent potential spread of infection but the methods used vary considerably. We designed a questionnaire which was piloted and then sent to 115 UK ENT out-patient departments to establish current UK practice. Most units (67 per cent) use a chemical soak system but the type of chemical disinfectant, the method of delivery and the duration of soak varied considerably. A few hospitals use a disposable plastic sheath system and others simply wipe the instrument with an alcohol swab in between patient use. The protocols for disinfection after high risk patients e.g. human immunodeficiency virus (HIV), hepatitis B varied from hospital to hospital. The results demonstrate a lack of standard practice that is wasteful of financial resources and may expose patients to unnecessary risk. There is a need for an authoritative ENT body to publish national guidelines as may be found in other specialties and other countries. PMID- 10829108 TI - Association of p53 expression with second primary tumour development in hypopharyngeal carcinoma. AB - Abnormalities of p53 tumour suppressor gene are detected in a diversity of malignancies and play an important role in their pathogenesis. Hypopharyngeal carcinoma is the most morbid among head and neck squamous cell carcinomas because of the high incidence of treatment failures and because a biological marker predictive of the treatment failures remains elusive. The expression of p53 protein in 46 hypopharyngeal squamous cell carcinomas was examined histochemically and p53 immunoreactivity was found in 19 of 46 cases (41.3 per cent). The rate of second primary tumour development was significantly higher in the p53-positive group than in the p53-negative group (p = 0.039), whereas that of tumour recurrence was not significantly different between the two. Moreover, there was no statistically significant difference in either overall or disease free survival between the p53-positive and -negative groups. These results indicate that although p53 expression significantly correlates with second primary tumour development in patients with hypopharyngeal squamous cell carcinomas, it is not predictive of the clinical outcome. PMID- 10829110 TI - A concha bullosa crusher for use in endoscopic sinus surgery. AB - A concha bullosa may require surgical reduction to facilitate access to the middle meatus during endoscopic sinus surgery. Here we describe an original instrument which enables this procedure to be performed simply with minimal mucosal damage. PMID- 10829111 TI - Middle-ear myoclonus. AB - Tinnitus produced by repetitive contraction of the middle-ear muscles is a rare condition. We present an interesting case of bilateral middle-ear myoclonus causing incapacitating tinnitus in a patient with multiple sclerosis. Otological examination demonstrated rhythmic involuntary movement of the tympanic membrane. These movements correlated with a rhythmic 'rushing wind' noise perceived by the patient. Oropharyngeal examination showed no evidence of palatal myoclonus. Impedance audiometry confirmed rhythmic change in the middle-ear volume. Medical management was unsuccessful. The patient's tinnitus was subsequently cured with bilateral sectioning of the tensor tympani and stapedial tendons. PMID- 10829112 TI - First branchial cleft sinus presenting with cholesteatoma. AB - Disorders of the first branchial cleft are rare and often present difficulties in diagnosis. A case is reported that presented with the clinical and histological features of cholesteatoma. The embryology, diagnosis and treatment of branchial arch defects are also reviewed. The case highlights the fact that a high index of suspicion should be aroused for patients with a discharging ear in the presence of an intact tympanic membrane. PMID- 10829113 TI - Herpes zoster oticus following mandibular block. AB - Although a few cases of facial palsy following mandibular nerve block and dental surgery have been described, it would appear that herpes zoster oticus following dental surgery has not been documented. It is possible that the latent virus may be activated by the mandibular nerve block and dental surgical interventions. Two cases of herpes zoster oticus, both following inferior alveolar nerve block anaesthesia for dental treatment are presented. PMID- 10829114 TI - Osteosarcomatosis involving craniofacial bones presenting with cranial nerve palsies. AB - Osteosarcomatosis (also known as multifocal osteosarcoma or multiple sclerotic osteosarcoma) is a rare condition, characterized by multiple skeletal lesions at initial presentation. We describe a case of osteosarcomatosis involving craniofacial bones presenting with cranial nerve palsies. In addition, a brief review of the literature is given, with a discussion of this highly aggressive form of osteosarcoma. PMID- 10829115 TI - Nasal lipoma. AB - Several lipomas of the nasal cavity and paranasal sinus have previously been reported to date. We present a 21-year-old case with nasal lipoma situated on the left posterior of part of the nasal septum, operated on endoscopically. The present case is, to our knowledge, the first report on nasal lipoma in the adult. PMID- 10829116 TI - Abductor vocal fold palsy as a manifestation of type one Arnold Chiari malformation. AB - We report a rare case of type 1 Chiari malformation which presented to the Otolaryngology department with abductor vocal fold palsy following an aspiration pneumonia secondary to absent laryngeal sensation. Chiari malformations and related cases are discussed. PMID- 10829117 TI - Vallecular cyst: report of four cases--one with co-existing laryngomalacia. AB - Congenital vallecular cysts are rare. In this report, four infants having vallecular cysts encountered over a six-year period from 1992 to 1997 were reviewed. All of them presented with upper aerodigestive tract symptoms. Marsupialization was performed in three of them and CO2 laser excision was performed in the fourth patient. There was no recurrence of the cyst in any patient. One of them also had co-existing laryngomalacia. The degree of airway collapse caused by laryngomalacia improved after cyst removal. The laryngomalacia resolved spontaneously. Cyst fluid culture was performed in one of the patients and yielded Staphylococcus aureus but there was no other definite indicator of infection. Staphylococcus aureus could also be isolated in the respiratory tract from two of the other patients. PMID- 10829118 TI - Retropharyngeal abscess as a complication of oropharyngeal trauma in an 18-month old child. AB - An 18-month-old boy presented to the accident and emergency department following trauma to the oropharynx by a doll's umbrella. Although no significant injury could be identified at first, it later transpired that he had developed retropharyngeal surgical emphysema with abscess. This case report aims to review the nature of retropharyngeal abscesses and to highlight and re-emphasize the fact that apparently non-serious pharyngeal injuries in children should be treated with suspicion. PMID- 10829119 TI - Metastatic endometrial carcinoma of the neck. AB - We report a case of metastatic endometrial carcinoma of the neck. A patient with a past medical history of squamous cell carcinoma of the larynx, breast carcinoma and endometrial carcinoma presented with a neck mass. Fine needle aspiration cytology (FNAC) showed this to be a poorly differentiated carcinoma with squamoid features and thus a potentially curative neck dissection was performed. Histology of the mass showed a clear cell endometrial carcinoma. Metastatic gynacecological malignancies to the head and neck are rare and this is the first reported case of metastatic endometrial carcinoma in the neck. PMID- 10829120 TI - Neural fibrolipoma: an unusual case. AB - A 45-year-old gentleman presented with a diffuse left neck mass. Surgical exploration revealed a large lipomatous lesion. Histological examination identified this to be a neural fibrolipoma. This is the first reported case of this lesion in the neck. PMID- 10829122 TI - The JLO Travelling Fellowship 1999 report. Visit to Professors van den Broek and Cremers, University Hospital Nijmegen, The Netherlands and Professor U. Fisch, UniversitatsSpital Zurich, Switzerland. PMID- 10829121 TI - Optimum imaging for mucoceles. AB - A mucocele is an epithelial lined mucus-containing sac completely filling a paranasal sinus and capable of expansion. They are relatively unusual, occurring most frequently in the fronto-ethmoidal region. The imaging features on plain X ray, computerized tomography and magnetic resonance imaging are relatively characteristic allowing distinction of the lesion from other pathologies in this area although the mucoceles may occur in association with other pathologies such as nasal polyposis and neoplasia. PMID- 10829123 TI - The catechism effect: child testimonies during a 17th-century witch panic as related to educational achievement. AB - Testimonies from 488 children given to the priests of the parish of Rattvik during a preliminary investigation of a Swedish witch panic in 1670-71 are examined in relation to records from parish catechetical meetings held in 1671. The result implies that children who knew and understood at least parts of Luther's catechism were less liable to have falsely alleged that they had been kidnapped by female satanists during the witch panic of the previous year. It is suggested that these effects were caused by differences in cognitive, social, and emotional resources among these children as compared to those who were unable to learn and understand any parts of Luther's catechism. PMID- 10829124 TI - The effects of stimulus set size and word frequency on verbal serial recall. AB - Two experiments are reported which examine immediate serial recall for high- and low-frequency words. The words in each list were either repeatedly drawn from the same small pool of candidates (in the closed set conditions) or each word only ever occurred once during the experiment (in the open set conditions). The results consistently show an effect of word frequency but the effect of set size was only apparent for low-frequency words. It is argued that both frequency and set size effects reflect processes concerning the "clean-up" of degraded short term memory traces. PMID- 10829125 TI - Disturbed memory and amnesia related to intensive care. AB - Patients, when admitted to an intensive care unit (ICU), have one thing in common: their illness is life-threatening. Patients may remain on ICU in a critical condition, needing support with their breathing, circulation, and/or kidneys for varying lengths of time, from days to weeks. During that time the patients will receive sedative and analgesic drugs to ensure compliance with artificial ventilation. Patients recovering from critical illness frequently have little or no recall of their period in ICU, or remember nightmare, hallucinations, or paranoid delusions. The nature, extent and reason for these difficulties, have been under-reported and consequently our purpose was to conduct a review of memory problems experienced by ICU patients. A systematic literature review of computer databases (Medline, PsycLit, and CINAHL) identified 25 relevant papers. In addition, other relevant articles were obtained, citation lists and associated articles retrieved. Due to lack of research on processes underlying memory problems in ICU patients all articles that introduced an insight into possible mechanisms were included in the review. There seem to be two possible processes contributing to memory problems in ICU patients. First the illness and treatment may have a general dampening effect on memory. Delirium and sleep disturbance are both common in ICU patients. Delirium can result in a profound amnesia for the period of confusion. Sleep deprivation exacerbates the confusional state. Slow wave sleep is important for the consolidation of episodic memories. Treatment administered to patients in ICU can have effects on memory. Opiates, benzodiazepines, sedative drugs such as propofol, adrenaline, and corticosteroids can all influence memory. In addition, the withdrawal of drugs, such as benzodiazepines, can cause profound withdrawal reactions, which may contribute to delirium. Second, we hypothesise that there is a process that affects memory negatively for external events but enhances memory for internal events. The physical constraints and social isolation experienced by ICU patients and the life-threatening nature of the illness may increase the experience of hypnagogic hallucinations. Attentional shift during hypnagogic images from external stimuli to internally generated images would explain why ICU patients have such poor recall of external ICU events, but can clearly remember hallucinations and nightmares. Patients describe these memories as being very vivid and this is explored in terms of flashbulb memory formation. The absence of memories for real events on ICU can result in ICU patients remembering paranoid delusions of staff trying to kill them, with little information to reject these vivid memories as unreal. This has implications for patients' future psychological health. PMID- 10829126 TI - Differential recall of central and peripheral details of emotional slides is not a stable phenomenon. AB - A number of studies have reported that central information of an emotional scene is well retained, whereas peripheral details of such a scene are poorly recalled. Experiment 1 tested the hypothesis that attentional narrowing is responsible for this phenomenon. In addition, an attempt was made to increase the ecological validity of the experiment by giving extensive self-relevant instructions. Results showed that, although an emotional slide elicited eye-movements consistent with attentional narrowing, the corresponding recall patterns were absent. Experiments 2 and 3 explored some of the variables that might be responsible for the latter result. Experiment 2, relying on the original design of Christianson and E.F. Loftus (1991), found enhanced recall of central information of an emotional scene. Experiment 3 systematically varied stimulus exposure and interstimulus interval durations. However, the results of this experiment were rather complex and did not fully support the predicted differential recall patterns. Possible explanations for these findings are discussed. It is suggested that other methods (e.g. increasing levels of emotion rather than involvement) may be more suitable for testing the attentional narrowing hypothesis of emotional memory. PMID- 10829127 TI - When did Mrs Thatcher resign? The effects of ageing on the dating of public events. AB - Two experiments investigated whether, in light of the commonly reported phenomenon of subjective time acceleration with age, there would be an effect of age on the dating of public events. In the first experiment covering the past seven years there was the suggestion of a decrease in forward telescoping with age, and in the second covering the period from 1977-89 this trend was continued, with the over-60s group now showing a tendency to date events too distantly. This effect is uncommon in dating studies and may offer evidence for the existence of time acceleration. An additional finding was that adults in the age range 35-50 years showed greater accuracy in dating events than did university students and adults over the age of 60. PMID- 10829128 TI - When false memories do not occur: not thinking of the lure or remembering that it was not heard? AB - The aim of the present study was to evaluate two explanations for the non occurrence of false memories in the Deese-Roediger-McDermott (DRM) paradigm. One explanation was that a critical lure is not recalled because the list failed to evoke it in the participant's mind. Another possible explanation was that the participant would identify the critical lure and would remember, at the time of recall, that the lure was not produced by an external source. In order to explore these two possible explanations for the non-occurrence of false memories, an experimental phase was added to the usual DRM paradigm: participants were asked to recall items they thought of but did not recall because these items were not members of the list presented by the experimenter. Among participants who did not recall the critical lure during the standard recall task, those who recalled the critical lure during the additional phase outnumbered those who did not recall it. This result is more consistent with the second explanation than with the first one. PMID- 10829129 TI - The prevalence of ocular defects and the provision of eye care in adults with learning disabilities living in the community. AB - Most people with learning disabilities now live in the community rather than in institutions, and community services need to be targeted appropriately. Vision screening was carried out in community Day Care Centres for adults with learning disabilities. One hundred and fifty four subjects took part, and screening was successful in 148 subjects, using retinoscopy and standard acuity tests, including preferential looking. The previously reported high prevalence of refractive errors, strabismus and pathological defects amongst people with learning disabilities was confirmed. Over 60% of subjects had below-normal distance acuity, which in many cases was exacerbated by uncorrected refractive errors. In all, 41% of subjects could have benefited from distance spectacles, and 56% from spectacles for near tasks. The lack of adequate spectacles was particularly high amongst adults with more severe learning disabilities. The study highlights the current lack of adequate eye care for people with learning disabilities, and demonstrates that eye examinations are possible for the majority of individuals. There is a need for greater awareness amongst families/carers and amongst optometrists to address this important issue. PMID- 10829130 TI - The practical near acuity chart (PNAC) and prediction of visual ability at near. AB - OBJECTIVE: To improve on present reading chart designs, providing a quick and accurate method to measure the near acuity threshold, of particular importance with low vision patients. DESIGN: The Practical Near Acuity Chart (PNAC) uses a single paragraph with 3 simple related words on each line (12 lower case letters). The line print size decreases in a logMAR progression (N80-N5). METHODS: The time taken to measure near acuity of 53 subjects aged 9-91 years with the PNAC and Bailey-Lovie near chart was recorded and compared to their distance acuity (Bailey-Lovie chart), contrast sensitivity (Melbourne Edge Test) and ability to read extracts of newsprint. RESULTS: There was no difference in near acuity threshold using related or un-related words. There was a high correlation (r = 0.97) between near acuity measured with the PNAC and Bailey Lovie charts. However, the time taken to measure near acuity was significantly faster with the PNAC (32 +/- 2 s vs 76 +/- 4 s, p < 0.001). Near acuity measured with either chart was highly correlated (p < 0.001) to distance acuity (r = 0.74), contrast sensitivity (r = 0.62) and ability to read newsprint (r = 0.87). The PNAC was shown to have high test-reliable (r = 0.99). CONCLUSIONS: The PNAC offers a quick but accurate way to measure near acuity and shows a high degree of correlation with distance acuity, contrast sensitivity and the ability to read newsprint. PMID- 10829131 TI - The relation between accommodative facility and general binocular dysfunction. AB - This study was designed to evaluate the relation between accommodative facility and accommodative and binocular dysfunctions. We determined whether failure to achieve 8 cycles per minute of binocular accommodative facility or 11 cycles per minute of monocular accommodative facility tends to be associated with these dysfunctions. Forty eight subjects, ages 10-30 years were examined and classified into four groups: 13 subjects with accommodative dysfunctions, 11 patients with binocular dysfunctions, 12 subjects with accommodative and binocular dysfunctions and 12 control subjects with refractive errors but no accommodative or binocular anomalies. Monocular and binocular accommodative facility was conducted using +/- 2.00 D flip lenses. In general, statistical analysis indicates that subjects with binocular and accommodative (ocular motor) anomalies performed significantly poorer than subjects of normal group on monocular and binocular facility tests. Monocular accommodative facility results showed more information about the dysfunction of the patient compared with the results of the binocular accommodative facility. In general data supported a relation between reduced accommodative facility and a general binocular dysfunction (accommodative or binocular) which demonstrates the importance of the accommodative facility test in diagnosing an accommodative or binocular anomaly. PMID- 10829132 TI - Macular pigment optical density measurement: a novel compact instrument. AB - A compact device to derive the optical density of human macular pigment (MP) using heterochromatic flicker photometry is described. The validity of the system is assessed by measuring the optical density spectra of MP in 12 healthy subjects and comparing this with well-established previously published values. The mean spectral absorbance characteristics of MP across subjects corresponds well with accepted values. As reported in other studies, our measurements show a wide variation of MP optical densities between individuals. In our technique within subject variability is low; standard deviations are between 0.025 and 0.15 in most cases. The overall optical density of MP ranged from 0.08 to 0.84 with a mean of 0.496 and standard deviation of 0.257 at 460 nm. The stimulus size was 0.95 degrees. The unique feature of the technique is that it allows free viewing (not Maxwellian View) of the stimulus, it can be conducted easily and quickly and does not need frequent re-calibration. PMID- 10829133 TI - Estimation of projection errors in human ocular fundus imaging. AB - Photogrammetric analysis of features in human ocular fundus images is affected by various sources of errors, for example aberrations of the camera and eye optics. Another--usually disregarded--type of distortion arises from projecting the near spherical shape of the fundus onto a planar imaging device. In this paper we quantify such projection errors based on geometrical analysis of the reduced model eye imaged by a pinhole camera. The projection error found near the edge of a 50 degrees fundus image is 24%. In addition, the influence of axial ametropia is investigated for both myopia and hyperopia. The projection errors found in hyperopia are similar to those in emmetropia, but decrease in myopia. Spherical as well as ellipsoidal eye shapes were used in the above calculation and their effect was compared. Our results suggest that the simple spherical eye shape is sufficient for correcting projection distortions within a range of ametropia from -5 to +5 diopters. PMID- 10829134 TI - Corneal shape of Hong Kong-Chinese. AB - Corneal asphericity (the p-value, p) and apical corneal radius (r0) were calculated for 63 Hong Kong-Chinese (39 male and 24 female), using the Topographic Modeling System (TMS-1). The values of p and r0 in, and between, the two principal meridians were compared, and the effects of refractive error and gender were also investigated. The mean +/- SD r0 and p along the flattest meridian of the right eye were 7.82 +/- 0.26 mm and 0.78 +/- 0.12 respectively. The mean +/- SD r0 and p along the steepest meridian of the right eye were 7.64 +/- 0.26 mm and 0.83 +/- 0.15 respectively. Meridional variations were found in r0 and in p. There was no correlation between p and r0 along the two principal meridians. p was not significantly correlated to the refractive error but r0 was. Our results agree with previous findings that while males have longer r0 than females, their p are not significantly different. PMID- 10829135 TI - Duplication of unique optical effects of ancient Egyptian lenses from the IV/V Dynasties: lenses fabricated ca 2620-2400 BC or roughly 4600 years ago. AB - Ancient (in fact, first known) lenses (ca. 4600 years ago) mainly from the IVth and Vth Dynasties of Egypt had truly remarkable and unique optical properties. These were parts of equally fascinating eye structures. These structures were fabricated as separate assemblies for insertion into funerary statues during certain brief windows of time (roughly from 2620-2400 BC, and 1750-1700 BC). These "eyes" appear to follow the observer as he/she rotated in any direction about these statues. In this paper, by simple means, we have recreated the optical properties of these unique lenses in the laboratory in order to help understand their special properties. PMID- 10829136 TI - Evidence that a gender difference in intraocular pressure is present from childhood. AB - Routine assessment of intraocular pressure (IOP) is usually confined to the clinical population aged over 40 years, i.e., to those individuals considered to be most at-risk of developing glaucoma. Published IOP data collected from assumed non-glaucomatous adult subjects have shown a gender difference in this parameter, with human females recording a consistently higher value than age-matched males. A recent study has also demonstrated that a diurnal variation in IOP can be recorded in adults across normal consulting hours. IOP material is presented here, as collected in the early afternoon (14.00-14.59 hours) from n = 140 school children aged > or = 9 < 12 years. It is complemented by previously published data, also recorded between 14.00-14.59, from n = 100 middle-aged adults aged 40 59 years, and n = 102 mature adults aged over 60 years. Analysis of these three data sets indicates that a non-statistically significant gender difference in mean IOP (female > male) is present from childhood; it also suggests that, for either gender, the absolute mean level of IOP remains little altered from childhood into and throughout adulthood. However the physiological basis for these observations remains obscure. PMID- 10829137 TI - Evidence for mild blue-yellow colour vision deficits immediately following fluorescein angiography. AB - AIMS: We have investigated the short term effects of fluorescein angiography on the blue-yellow, red-green, and luminance contrast sensitivity of patients with early age-related macular degeneration (ARMD). METHODS: Nine ARMD patients with no exudative complications and a visual acuity of 20/60 or better in the tested eye were selected. Cardinal colour directions for the isolation of the red-green, blue-yellow and achromatic (luminance) visual mechanisms were determined for each patient. Contrast sensitivity was measured in each cardinal colour direction immediately before and 20 min after standard 20-flash fluorescein angiography. RESULTS: A significant, albeit mild, reduction for blue-yellow contrast sensitivity following angiography was observed (ANOVA, alpha = 0.05). Fluorescein angiographic exposure had no significant effect on red-green or luminance contrast sensitivity. CONCLUSION: Our results show that fluorescein angiography causes at least a short term deficit selective to blue-yellow contrast sensitivity in our patient group. PMID- 10829138 TI - Letter legibility and chart equivalence. AB - INTRODUCTION: We tested the appropriateness of the assumption that charts, which only vary in having different sequences of five letters per line, chosen from the same 10 letter set, can be regarded as equivalent for the purpose of making valid comparisons of visual acuity. METHOD: Visual acuity findings from samples of 400 patients, for each of two nominally equivalent Bailey-Lovie charts, have been used to determine the relative legibility of individual letters and lines, using the percentage incorrect method of analysis. RESULTS: The chance of error for the hardest letter is approximately 13 times greater than for the easiest letter on each chart. Significant between-chart differences in error rates for the hardest letters ('F' and 'H') were found. Some letters that have adjacent ranks on the legibility scale were found to have significantly different legibility. Significant differences in difficulty can occur for the same nominal line on apparently equivalent charts because of chance combinations of easier or harder letters in that line. Uneven line-to-line scale intervals have been confirmed for the charts examined, by showing corresponding differences in the distributions of lines of threshold acuity. CONCLUSIONS: The use of varying examination distances may be inappropriate for Bailey-Lovie (or similar) charts that depart from their nominal interval scaling. When different versions of these charts are assumed to be equivalent, the discrepancy between repeated measurements may be significantly increased. There is the possibility of increasing measurement precision using charts having different sequences of the same 10 letters in each line to achieve equal scaling of line intervals. Equivalent charts can be validly constructed using different sequences of the same 10 letters in each line. PMID- 10829139 TI - Intra-observer repeatability and inter-observer agreement of the Smith method of measuring the anterior chamber depth. AB - The Smith (1979) method provides a means of estimating the anterior chamber depth without additional attachments to the slit lamp [Smith, R. J. H. (1979). A new method of estimating the depth of the anterior chamber. Br. J. Ophthalmol. 63, 215-220]. In this study, the 95% intra-observer limits of repeatability and the 95% inter-observer limits of agreement of this method have been determined. The intra-observer limits of repeatability were determined by plotting the difference vs the mean of the estimated anterior chamber depth obtained in two different sessions by each of two examiners, while the inter-observer limits of agreement are represented by a plot of the difference vs the mean estimated anterior chamber depth between the two examiners. For one examiner, the 95% intra-observer limits of repeatability was -0.36 to 0.58 mm, while for the other examiner the 95% intra-observer limits of repeatability was -0.25 to 0.37 mm. The 95% inter observer limits of agreement were -0.31 to 0.23 mm and -0.41 to 0.25 mm for the first and second sessions respectively. The intra-observer limits of repeatability are comparable with those reported for A-scan ultrasonographic measures of the anterior chamber depth. These results imply that the Smith method can be used with a high degree of repeatability and agreement to clinically monitor longitudinal changes in anterior chamber depth. PMID- 10829140 TI - Precision of autokeratometry expressed as confidence ellipses in Euclidian 2 space. AB - A number of studies have examined the repeatability or precision of keratometers. All studies have found exceptionally high precision in keratometry of the spherically equivalent proportion, while the joint variation in astigmatic direction and magnitude barely has been explored. The purpose of this study was to examine the simultaneous variation in astigmatic direction and magnitude between two single autokeratometries, using a Nidek ARK 2000-S autokeratorefractometer. A single autokeratometry was performed on two occasions on 50 right eyes of patients with cataracts. The precision was defined as the paired difference between these two measurements. Each net astigmatism was transformed to polar values, and uni- and bivariate polar value analysis was performed. Precision of autokeratometry, expressed as a mean difference and a 95% confidence area, was calculated as a confidence ellipse in Euclidian 2-space. The accompanying net astigmatisms with confidence perimeters were derived by point for-point transformation from polar values. Clinical variability may overwhelmingly be attributed to patient-related factors and only to a small degree to autokeratometry itself. Clinicians should know the precision of their keratometers, both for daily use and in the planning and dimensioning of scientific studies. Our results indicate that a minimum number of 25 eyes in each group is necessary in order to detect a difference of 0.25 diopters in surgically induced astigmatism between two surgical techniques. Precision of keratometry can only be determined from polar values or similar entities and never by the use of net astigmatisms. PMID- 10829141 TI - Meredith W. Morgan, 1912-1999, Professor of Physiological Optics and Optometry, Dean of the School of Optometry, University of California at Berkeley. PMID- 10829142 TI - Who are we here for? PMID- 10829143 TI - The level of need for palliative care: a systematic review of the literature. AB - Palliative care services have developed rapidly over the past 30 years, with little evaluation as to how needs have been met by these new services. As part of a systematic review of palliative care, evidence of the needs of patients and carers has been evaluated from the current literature. Of the total of 673 articles related to the 10 areas within the main review, 64 provided evidence on the need for palliative care services over the period from 1978 to 1997. A further nine articles were added in November 1998 after the end of the study of update the review with more recent research. Need can be assessed in one of two ways: either by adopting an epidemiological approach or by examining health service usage. In the former, evidence is provided on disease-specific mortality, and related to the duration of symptoms prior to the patient's death. As an example of this, it is suggested that services may need to provide pain control for 2800 patients per million (p/M) population dying from cancer each year and 3400 p/M with noncancer terminal illness. Using health service usage as an indicator of need, 700-1800 p/M with cancer and 350-1400 p/M with noncancer terminal illness would require a support team or specialist palliative home care nurse, with 400-700 cancer p/M and 200-700 noncancer p/M requiring inpatient terminal care. Studies indicate that at present usage, palliative care is being provided by 40-50 hospice beds/M. Despite this provision, there remains evidence that in certain areas of care such as pain control, there still remains a high degree of unmet need. PMID- 10829144 TI - Telling the diagnosis to cancer patients in Japan: attitude and perception of patients, physicians and nurses. AB - The aim of this study was to clarify patients', physicians', and nurses' perceptions with regard to the communication of diagnosis to cancer patients in Japan. Sixty-three cancer patients, 35 physicians and 21 nurses were enrolled for this study: 54 of the patients wished to be informed of the diagnosis, of whom 34 had actually been told that they had cancer. Physicians did not tell the truth to the remaining 20 patients, of whom seven were not told the diagnosis because family members objected. Twenty-one of the 35 physicians thought that telling the true diagnosis had a positive effect and 27 thought that disclosure of the diagnosis to cancer patients should be promoted. Sixteen of the 21 nurses did not experience any difficulties with patient care after the diagnosis was disclosed. The present study suggests that medical staff and family members should respect the patient's standpoint because patients have the right to know about their own condition. Physicians should first provide the details of the disease to their patients. Thereafter, family members should be informed, but only with the patient's consent. PMID- 10829145 TI - Constipation and the use of laxatives: a comparison between transdermal fentanyl and oral morphine. AB - Constipation and the use of laxatives were investigated in patients with chronic cancer pain treated with oral morphine and transdermal fentanyl in an open sequential trial. Forty-six patients were treated with slow-release morphine 30 1000 mg/day for 6 days and 39 of these patients were switched to transdermal fentanyl 0.6-9.6 mg/day with a conversion ratio of 100:1. Median fentanyl doses increased from 1.2 to 3.0 mg/day throughout the 30-day transdermal treatment period. Twenty-three patients completed the study. Two patients died from the basic disease while treated with transdermal fentanyl, 12 patients were excluded for various reasons, and not enough data for evaluation were available for two patients. Mean pain intensity decreased slightly after conversion although the number of patients with breakthrough pain or requiring immediate-release morphine as a rescue medication was higher with transdermal fentanyl. The number of patients with bowel movements did not change after the opioid switch but the number of patients taking laxatives was reduced significantly from 78-87% of the patients per treatment day (morphine) to 22-48% (fentanyl). Lactulose was used mainly and was reduced most drastically, but other laxatives were also used less frequently. In this study transdermal fentanyl was associated with a significantly lower use of laxatives compared to oral morphine. The difference in the degree of constipation between the two analgesic regimens should be confirmed in a randomized double-blind study that takes into account both constipation and use of laxatives. PMID- 10829146 TI - Rehabilitation for elderly patients with cancer asthenia: making a transition to palliative care. AB - The transition from curative to palliative care for elderly patients with cancer related asthenia presents complex challenges to oncologists. A result of this complexity is a lack of regard for the maintenance of physical and mental function. This study examined the effects of comprehensive, multidisciplinary, inpatient rehabilitation on the physical and mental function of elderly cancer patients with asthenia. We found that both physical and mental function improved following inpatient rehabilitation. However, it is difficult to predict which patients benefit most from rehabilitation. Nevertheless, functional improvement following rehabilitation may allow medical caregivers to better distribute the type and intensity of care, and reduce the caregiver burden experienced by family members in the home. PMID- 10829147 TI - Macmillan Carers Schemes in England: results of a multicentre evaluation. AB - Maintaining sick and elderly people at home, particularly as they approach the end of life, is a long-established challenge for health and social care services. Over the past 30 years palliative care providers have attempted a variety of innovations in this area. We report on a descriptive study of seven pilot Macmillan Carers Schemes in England. The schemes sought to provide practical and emotional help to cancer patients and families living in their own homes. Data are available on 624 referrals to the schemes over a 1-year period. Emphasizing comparisons between schemes, we report on reasons for and sources of referral, services offered, number and duration of visits and tasks undertaken. We consider the views of informal carers who made use of the service, the perceptions of Macmillan carers themselves, and we compare financial costs of schemes. It is concluded that the schemes have the potential for further development but face problems, which reflect on their borderline position between 'health' and 'social care'. Current policy changes may be beneficial to the schemes in this respect. PMID- 10829148 TI - Palliative care, assisted suicide and euthanasia: nationwide questionnaire to Swedish physicians. AB - The objective of this study was to investigate what actually happens between physicians and adult patients in difficult end-of-life situations. We circulated an anonymous questionnaire to a randomized sample of 952 Swedish physicians registered in specialties comprising care of dying adult patients, 122 palliative care physicians, and 130 physicians from the Swedish Association for the Study of Pain. Of special interest were themes in conversations between the physicians and the patients, desires expressed by the patients, and actions performed by the physicians that might affect the patients' expected survival. The overall response rate was 79%. Of these, 63% of the randomized physicians, 95% of the palliative care physicians, and 43% of the Association for the Study of Pain physicians had more than occasionally treated dying adult patients during the past year. About half of them had discussed palliative care with all their dying patients, and more than half of the physicians had heard their patients expressing a wish to die. About one-third of all the physicians had given analgesic or other drugs in such doses that some of their patients' deaths were hastened. The same proportion had also been asked for active euthanasia, while 10% had been asked to assist suicide. No case of euthanasia and only a few cases of assisted suicide were reported. By implication, the study suggests that improving patients' awareness of the possibilities to relieve pain, anxiety and dyspnoea during the final days of life is an important way to reduce requests for active euthanasia. PMID- 10829149 TI - Spirituality and palliative care: a model of needs. AB - This paper provides a theoretical model of spiritual needs in palliative care based on a review of the palliative care literature. Three sources of transcendence, the building blocks of spiritual meaning, are identified: the situational, the moral and biographical, and the religious. After areas of transcendence are described and explained, implications for future theory, research and practice are identified. PMID- 10829150 TI - Leeds eligibility criteria for specialist palliative care services. PMID- 10829152 TI - Do home deaths increase distress in bereavement? PMID- 10829151 TI - Palliative care services: views of terminally ill patients. PMID- 10829153 TI - Autonomy for whom? A perspective from the Orient. PMID- 10829154 TI - An aphorism on afterlife is medically unreasonable. PMID- 10829155 TI - Converting from oral morphine to subcutaneous diamorphine. PMID- 10829156 TI - Gabapentin for treatment of thalamic pain syndrome. PMID- 10829157 TI - Planning and management of the difficult primary hip replacement: preoperative planning and technical considerations. PMID- 10829158 TI - Difficult primary total hip arthroplasty: selected surgical exposures. PMID- 10829159 TI - Primary total replacement of the dysplastic hip. PMID- 10829160 TI - The bearing surface in total hip arthroplasty: evolution or revolution. AB - Highly cross-linked polyethylenes represent a new class of polyethylenes that demonstrate dramatic improvements of wear characteristics in laboratory tests. Cross-linked polyethylenes can be manufactured by a number of methods, all of which lead to some changes in the physical properties of the polyethylene. The very limited clinical information about cross-linked polyethylenes available has been favorable. Cross-linked polyethylenes appear to hold promise as an alternative to conventional polyethylene and to hard-on-hard bearing surfaces, but much more clinical information will be required before they can be recommended for routine clinical use. PMID- 10829161 TI - The difficult socket. PMID- 10829162 TI - The difficult femur. PMID- 10829163 TI - Osteolysis: etiology, prosthetic factors, and pathogenesis. PMID- 10829164 TI - Femoral bone loss in total hip arthroplasty: classification and preoperative planning. AB - A single comprehensive classification system that can adequately describe all types of bone loss associated with hip arthroplasty should become a standard for reporting purposes. There is a need for a critical appraisal of the classification systems currently in use and, through a consensus, for development of a system that will permit comparison between the reported results of different techniques. Although no one classification system is ideal, the one proposed by the AAOS Committee on the Hip is the most comprehensive and the most consistently used. It addresses not only revision total hip arthroplasty but also primary hip replacement. It also addresses other conditions related to problems with the bone stock, such as those resulting from a previous hip arthrodesis on the acetabular side and femoral stenosis and malalignment on the femoral side. The only drawback to this classification system is its complexity; however, the problem of acetabular and femoral bone loss is of sufficient complexity and variety that a simple classification system, although ideal, cannot be comprehensive. Regardless of the absence of a common language and a comprehensive classification system that is applicable to all types of reconstructions, it is clear that femoral bone loss is a problem that will continue to challenge orthopaedic surgeons. It is only by careful and methodical analysis of patients who have femoral bone loss and by meticulous attention being paid to detail in preoperative evaluation and investigation, surgical planning, and the recording of outcomes that we will be able to improve our treatment of this difficult problem. PMID- 10829165 TI - Basic science of onlay allografts: a review. PMID- 10829166 TI - The basic science of impaction allografting in revision hip surgery. PMID- 10829167 TI - Current status of impaction allografting for revision of a femoral component. PMID- 10829168 TI - Hip replacement: treatment of femoral bone loss using distal bypass fixation. PMID- 10829169 TI - Fixation with a modular stem in revision total hip arthroplasty. PMID- 10829170 TI - Proximal femoral replacement with megaprosthesis. PMID- 10829171 TI - Femoral bone loss in patients managed with revision hip replacement: results of circumferential allograft replacement. PMID- 10829172 TI - Total hip arthroplasty in patients with neuromuscular disease. PMID- 10829173 TI - Understanding and treating osteonecrosis of the femoral head. PMID- 10829174 TI - Basic biology of the meniscus and response to injury. AB - This brief description of the basic biology of the meniscus helps to demonstrate that the menisci are an integral part of the complex biomechanics of the knee. Without the menisci, the normal knee joint function is severely diminished. Lack of the menisci typically leads to progressive degenerative joint disease of the knees. Efforts must be continued to find better methods and techniques to manage meniscus injuries. Improved therapy will help minimize the late instability of the joint, decrease pain associated with loss of the meniscus, minimize or prevent degenerative joint changes, and obviate the need for multiple surgical procedures. Specific targets should include improved methods of meniscectomy, new and better techniques of repair for meniscus tears, methods to enhance the cellular response for healing of meniscus tissue, and, finally, new ways to regenerate lost or damaged tissue. PMID- 10829175 TI - Arthroscopic meniscal repair with use of the outside-in technique. AB - The outside-in technique of arthroscopic repair is effective for repair of most meniscal tears. The overall indications for the use of this technique are similar to those for the commonly used inside-out technique. The outside-in technique is especially useful for suturing the anterior horn of the meniscus as well as for suturing meniscal replacement devices such as a collagen meniscal implant or a meniscal allograft. Other specific advantages of this technique include the ability to predictably avoid neurovascular injury without the need for a large posterior incision. A particular disadvantage is the difficulty of achieving perpendicular orientation of sutures when a tear is adjacent to the site of attachment of the posterior horn. Use of the inside-out technique or an all inside implant is suggested for these tears. The use of this suturing technique is facilitated by attention to several technical points. The knee should be maintained in flexion for repair of tears of the lateral meniscus (to avoid injury to the peroneal nerve) and in nearly full extension for repair of the posterior aspect of the medial meniscus (to avoid injury to the saphenous nerve and its branches). Care must be taken to avoid tying the sutures around a branch of the saphenous nerve during repair of the medial meniscus. The sutures should be retrieved through a cannula in the anterior portal to avoid the entrapment of the sutures in soft tissue. A probe can be used to prevent displacement of the inner fragment of a bucket handle tear when the needles are placed across the tear, as the entering needles may push the torn fragment into the knee. A vertical suture orientation is preferred in order to evenly co-apt the meniscus to the capsule. If knot-end sutures (so-called Mulberry knots) are used, 2 sutures can be vertically stacked, with 1 on each surface of the meniscus. If a mattress suture is used, a vertical orientation is easily achieved with the outside-in technique. Use of an exogenous fibrin clot is suggested for isolated tears. The clot can be secured to the site of repair by a suture that has been placed through a spinal needle with the outside-in method. Delayed weightbearing should be considered as postoperative management for patients who have had repair of a tear with a radial component or repair of a complex tear in which a fibrin clot was used. Previous studies have demonstrated that the location of the tear and the condition of the anterior cruciate ligament are important factors in determining the success of meniscal repair. The overall results with use of the outside-in technique are comparable with those reported with use of the inside out method. Patients with concomitant tears of the medial meniscus and the anterior cruciate ligament should have combined meniscal repair and reconstruction of the anterior cruciate ligament. As healing was demonstrated in 8 of 13 patients with an unrepaired tear of the anterior cruciate ligament, consideration should still be given to meniscal repair in patients who refuse reconstruction of the anterior cruciate ligament. In this setting, it may be advisable to use multiple permanent sutures, and the patient must be counseled regarding the higher rate of failure with this approach. Repairs of the lateral meniscus have a higher rate of success, and repair of the lateral meniscus should be considered even in the presence of injury of the anterior cruciate ligament. PMID- 10829176 TI - Meniscal repair rehabilitation. PMID- 10829177 TI - Surgical treatment of arthrofibrosis of the knee. PMID- 10829178 TI - Techniques of wrist arthroscopy. PMID- 10829179 TI - Basics of elbow arthroscopy. AB - With more innovation in arthroscopic equipment and surgical technique, elbow arthroscopy will continue to evolve and new indications will emerge. Strict adherence to the principles outlined above will allow the use of arthroscopy to treat a variety of elbow disorders in a safe and effective manner. PMID- 10829180 TI - Arthroscopic treatment of arthritis of the elbow. PMID- 10829181 TI - Complications of elbow arthroscopy. PMID- 10829182 TI - Arthroscopic ankle arthrodesis. AB - The obvious socioeconomic advantages are quite dramatic, with early weight bearing and AFO immobilization allowing patients an early return to work. Outpatient surgery is a cost-effective benefit. Patient satisfactions as well as comfort are greatly enhanced, requiring only oral pain medication. All patients have tolerated their postoperative regimen and same-day discharge. Arthroscopic subtalar arthrodesis is a technically demanding procedure that requires some rather advanced arthroscopic skills to perform. Joint access is tight, restricted, and requires small instrumentation. Deformities cannot be corrected; therefore, at this stage, a fusion in situ must be considered. The learning curve is certainly far steeper because of the smaller patient population available for enhancing surgical skills. Overall, this procedure has stood the test of time and follow-up. The results appear to be excellent in terms of patient satisfaction, fusion rate time until union, and postoperative morbidity. The recognition and enhancement of this technique as well as the development of more advanced technology will certainly allow this arthroscopic subtalar arthrodesis technique to mature even further over time. PMID- 10829183 TI - Management of the diabetic stiff shoulder. PMID- 10829184 TI - Compressive radial neuropathies. AB - Radial neuropathy can have one of several clinical presentations, depending on the level of compression: high radial nerve palsy, PIN palsy, radial tunnel syndrome, and Wartenberg's syndrome. Elucidating the history and progression of symptoms with a physical examination directed at testing individual muscles will determine the approximate anatomic level of radial injury. Identification of a Tinel's sign will often locate the site of compression. Electrodiagnostic studies can confirm the diagnosis in a radial motor neuropathy, but are unreliable in radial tunnel syndrome and sensory radial nerve compression. A trial of nonsurgical treatment is warranted in all cases except those with progressive motor weakness or palsy. Patients who do not respond or continue to progress despite conservative treatment are candidates for surgical decompression. PMID- 10829185 TI - Compressive neuropathies of the ulnar nerve at the elbow and wrist. AB - Compressive neuropathy of the ulnar nerve in the upper limb is a common problem that frequently results in severe disabilities. At the elbow, Lundborg concluded that the nerve was "asking for trouble" because of its anatomic course through confined spaces and posterior to the axis of elbow flexion. Normally, the ulnar nerve is subjected to stretch and compression forces that are moderated by its ability to glide in its anatomic path around the elbow. When normal excursion is restricted, irritation ensues. This results in a cycle of perineural scarring, further loss of excursion, and progressive nerve damage. Initial treatment for the acute and subacute neuropathy at the elbow is nonsurgical. Rest and avoiding pressure on the nerve may suffice, but if symptoms persist, splint immobilization of the elbow and wrist is warranted. For chronic neuropathy associated with muscle weakness, or neuropathy that does not respond to conservative measures, surgery is usually necessary. A variety of surgical procedures have been described in the medical literature, and deciding on the most effective procedure can be difficult considering the excellent results claimed by proponents for each. Unfortunately, there is a paucity of information based on prospective randomized clinical studies comparing the different surgical methods. Dellon attempted to provide some guidelines by reviewing the data in 50 articles dealing with nonsurgical and surgical treatment of ulnar neuropathies at the elbow. In order to provide uniform data, he re-interpreted the data in these articles using his own system for staging nerve compression. He reported that treatment was most successful for mild neuropathies, a conclusion few would challenge. Excellent results were also achieved in 50% of patients with mild neuropathies that were treated nonsurgically and in more than 90% treated by surgery, regardless of the procedure. For moderate neuropathies, nonsurgical treatment was generally unsuccessful, as were decompressions in situ. Medial epicondylectomies were effective in only 50% of cases and they had the highest recurrence rate. Regarding ulnar nerve transpositions, each method has its proponents, usually based on the training and experience of the surgeon. Subcutaneous transposition is the least complicated. It is an effective procedure, particularly in the elderly and in patients who have a thick layer of adipose tissue in their arms. It is the procedure of choice for repositioning the nerve during surgical reductions of acute fractures, arthroplasties of the elbow, and secondary neurorrhaphies. Intramuscular and submuscular transpositions are more complicated procedures. Although proponents of intramuscular transposition report favorable results, the procedure can result in severe postoperative perineural scarring. Submuscular transposition has a high degree of success and is generally accepted to be the preferred procedure when prior surgery has been unsuccessful. I also prefer it as the primary procedure for most chronic neuropathies that require surgery. Compressive neuropathies of the ulnar nerve in the canal of Guyon are less common, but they can also result in significant disabilities. Compression can occur in 1 of 3 zones. Zone 1 is in the most proximal portion of the canal, where the nerve is a single structure consisting of motor and sensory fascicles, and zones 2 and 3 are distal where the ulnar nerve has divided into motor and sensory branches. The clinical picture correlates with the zone in which compression occurs. PMID- 10829186 TI - Tendon transfers about the shoulder and elbow in obstetrical brachial plexus palsy. PMID- 10829187 TI - Cervical degenerative disorders: etiology, presentation, and imaging studies. PMID- 10829188 TI - Cervical spondylotic myelopathy and radiculopathy. AB - Appropriate management of degenerative cervical spine conditions requires careful elucidation of the presenting clinical syndrome. Because of the pervasiveness of degenerative changes in asymptomatic patients, a clear correlation of symptoms, physical signs, and imaging findings is required before any specific diagnosis can be made. At this time, surgery is not recommended for prophylactic decompression in asymptomatic patients or in those patients with neck pain in the absence of extremity symptoms. In most patients with radiculopathy or mild myelopathy, a trial of nonsurgical management is recommended. Ultimately, patients with neurologic complaints and in whom nonsurgical measures have failed, as well as those with more pronounced myelopathy, should be offered surgical intervention. Selection of the safest, yet sufficient, approach requires a clear understanding of the benefits and expected outcomes. The outlook for patients with both cervical radiculopathy and early myelopathy is good. Radicular symptoms usually improve, but gait and hand changes may not. LF is preferred in younger patients with posterolateral or lateral soft disk herniations, or focal foraminal osteophyte impingement and predominance of upper extremity symptoms. More central 1- or 2-level pathology should be treated with ACDF. Anterior cervical corpectomy should be entertained in patients with nondisk level encroachment and in those with 3 contiguous levels of pathology. This approach is also required in cases of kyphosis and instability. Laminoplasty is indicated in patients with 4 or more levels of stenosis, particularly in those with global conditions such as continuous OPLL or congenital stenosis. In these patients, kyphosis or severe deformity may be addressed with a circumferential approach. PMID- 10829189 TI - Spinal stenosis. PMID- 10829190 TI - Treatment of the severely injured upper extremity. PMID- 10829191 TI - Sternoclavicular joint injuries and disorders. PMID- 10829192 TI - Complications of treatment of complete acromioclavicular joint dislocations. PMID- 10829193 TI - Orthopaedic treatment of fractures of the long bones and pelvis in children who have multiple injuries. PMID- 10829194 TI - Treatment of fractures complicated by burn or head injuries in children. PMID- 10829195 TI - The land mine epidemic. PMID- 10829196 TI - Soft-tissue masses: when and how to biopsy. AB - Patients with soft-tissue tumors present a diagnostic challenge to the clinician. A systematic approach to evaluation and biopsy is necessary to minimize diagnostic errors and biopsy complications. With MRI, the clinician and radiologist are able to precisely identify some soft-tissue masses, such as lipomas, ganglions, hemangiomas, and popliteal cysts. Most soft-tissue tumors have signal characteristics that are low signal on T1-weighted images and high signal on T2-weighted images. Soft-tissue masses that cannot be identified are called indeterminate masses, and a biopsy is necessary to identify these lesions. Needle biopsy and open incisional biopsy are effective methods to histologically identify the nature of the tumor. Open excisional biopsy should be performed only when the surgeon knows that the soft-tissue lesion is benign. PMID- 10829197 TI - New developments in the staging and imaging of soft-tissue sarcomas. PMID- 10829198 TI - Metastatic bone disease: evaluation, clinicopathologic features, biopsy, fracture risk, nonsurgical treatment, and supportive management. PMID- 10829199 TI - Surgical treatment for metastatic disease of the pelvis and the proximal end of the femur. AB - Advances in the treatment of invasive cancers continue to improve the longevity of patients who have these diseases; thus, the care of patients who have bone metastases is an issue of the utmost importance to the orthopaedic surgeon. In terms of maintaining the ability to walk, no site of potential metastatic involvement is more crucial than the proximal end of the femur and the acetabulum. Advances in femoral and acetabular implants, imaging modalities, and operative techniques now allow reconstruction of even the most complex acetabular and proximal femoral defects. However, the orthopaedic surgeon must recognize the need to approach management of these patients from a multidisciplinary perspective. The oncologist, radiotherapist, rehabilitation medicine specialist, radiologist, and pathologist each have a role to play. Only through cooperation among all members of the team will a patient who has metastatic disease or a myeloma be given the best possible care. PMID- 10829200 TI - Metastatic disease to the spine: diagnosis and treatment. PMID- 10829201 TI - Synovial joint degeneration and the syndrome of osteoarthritis. AB - The adverse effects of osteoarthritis on the quality of life for tens of millions of people and on the costs of health care and economic productivity make it a major health problem that will increase in incidence and impact with the aging of the population. Although in most patients the cause of osteoarthritis remains unknown and no cure has been identified, appropriate diagnosis and treatment, including education of the patients, can minimize symptoms and help patients maintain active and productive lives. Accomplishing this aim requires that physicians have an understanding of the pathophysiology of joint degeneration and the relationships between joint degeneration and the clinical syndrome of osteoarthritis. Although much remains to be learned about the development and progression of the joint degeneration responsible for osteoarthritis, it is clear that the loss of articular cartilage results from disruption of the structural integrity of the articular cartilage coupled with or caused by an imbalance in the anabolic and catabolic activity of the tissue. The progression of joint degeneration varies considerably among patients. In some individuals the joint degenerates rapidly; in others the degenerative changes progress slowly over decades and in others the disease may remain stable. In rare patients, the joint degenerative changes spontaneously improve. Although joint degeneration is the underlying cause of the symptoms of osteoarthritis, including joint pain and loss of joint function, not all patients with joint degeneration have symptoms of osteoarthritis. Future therapeutic approaches may include disease-modifying drugs and surgical procedures that correct mechanical abnormalities, debride joints, and replace degenerated articular cartilage with implants that stimulate restoration of a cartilaginous joint surface. Development of methods to detect and monitor subtle changes in cartilage metabolism and identify the joint changes that precede loss of articular cartilage may make it possible to detect the earliest signs of osteoarthritis, when therapeutic interventions have the greatest potential for preventing progression of the disease. PMID- 10829202 TI - New approaches to the treatment of osteoarthritis: oral glucosamine and chondroitin sulfate. PMID- 10829203 TI - Hyaluronic acid therapy. AB - A significant amount of evidence exists suggesting beneficial effects of intra articular hyaluronate preparations in treating the symptoms of OA. The beneficial effects of these compounds tend to persist for several months after cessation of therapy, unlike anti-inflammatory drug effects. In addition, several clinical trials comparing these agents to anti-inflammatory medications indicate efficacy of symptom improvement equivalent to that of the anti-inflammatories. A few clinical studies have failed to demonstrate an effect, and clearly much work remains to be done in this area to fully determine rational treatment strategies with these agents. The symptom improvement is generally moderate, and additive effects of these agents with anti-inflammatories have not been demonstrated. Adverse local reactions have been reported in some clinical series of hyaluronate injections. Although the invasive nature of the procedure is a drawback, overall it appears to be well tolerated and is probably not harmful to articular cartilage, a major advantage over the widely practiced introduction of intra articular steroids. Documentation of true chondroprotective effects or alteration of the natural history of cartilage degeneration by these agents is lacking in the clinical literature and awaits further study. Cost effectiveness of this therapy versus other treatments, including the costs of treatment of side effects, also requires further rigorous evaluation, and may influence acceptability of this form of treatment to various health care providers and organizations. PMID- 10829204 TI - Preoperative planning in deformity correction and limb lengthening surgery. AB - A simple method of preoperative planning in deformity correction and limb lengthening surgery is described and illustrated with two cases of deformity associated with length discrepancy. Frontal and lateral radiographs allow defining a deformity in its actual plane. Tracings are made, axes are marked, and the deformity is analyzed graphically. A problem list is created on a worksheet. The osteotomies are made with scissors on the worksheet and the segments are angulated into position to test the correction. The method of fixation is selected and the patient seen preoperatively to address issues to be encountered during the correction. PMID- 10829205 TI - Orthopaedic management of chronic pain: pain management for the cancer patient. AB - Chronic pain is best managed through a multidisciplinary approach, which includes traditional and nontraditional therapies. Orthopaedists probably deal with chronic pain patients more than any other medical specialists. As Foley points out, it is critical that a trusting relationship between the patient and the physician is established; a relationship in which the patient believes the physician is taking the pain symptoms seriously and intends to address the pain aggressively. Although dealing with chronic pain can be a very frustrating part of orthopaedic practice, it can also be seen as a challenge. PMID- 10829206 TI - Local and regional anesthesia for the management of pain in orthopaedic surgery. PMID- 10829207 TI - Pain management and medical hypnosis. PMID- 10829208 TI - Pain management and electromagnetic medicine. PMID- 10829209 TI - Reflex sympathetic dystrophy: alternative modalities for pain management. AB - For the patient presenting with early symptoms (< 6 months) I usually start treatment with a dose pack of methylprednisolone, analgesics, and daily occupational/physical therapy for 2 weeks (Fig. 2). If they do not respond within the first week, I add stellate ganglion blocks and acupuncture to the treatment regimen. For patients presenting with established chronic pain, I immediately start them on a dose pack of methylprednisolone for 1 to 2 weeks, a nonsteroidal anti-inflammatory such as indomethacin, 50 mg 3 times a day for 10 days and then switch to 75 mg twice daily until there is a response. Amitriptyline is helpful for sleep and depression and also has a beneficial effect on blood flow. Calcium channel blockers (nifedipine) may help improve peripheral circulation by its effect on vascular smooth muscle. In this patient group, I almost always start stellate ganglion blocks on the first visit. I have the patient try at least 2 blocks before deciding whether or not blocks are helpful. Many patients will not respond to the first block, but will start to respond after the second block. If the blocks are helping, I recommend 3 blocks a week, every other day for 3 weeks. Patients get the most benefit from their blocks if they have occupational or physical therapy immediately following the block. Surgical sympathectomy may be helpful but only in patients who have responded to sympathetic blockade. PMID- 10829210 TI - Acupuncture: alternative modalities for pain management. PMID- 10829211 TI - Orthopaedic surgery in the developing world: an introduction. PMID- 10829212 TI - Volunteer opportunities for orthopaedists in the developing world. PMID- 10829213 TI - The transfer of appropriate orthopaedic technology to less developed countries: a philosophy of involvement. PMID- 10829214 TI - Selected conditions common in the developing world. AB - Although many esoteric diseases occur in the developing world, the majority of the orthopaedic problems encountered are similar throughout the world. Most are secondary to trauma, infection, or congenital anomalies. The geographic differences in provision of care are related to timing of treatment and available facilities. The information presented in this chapter offers examples of successful treatment regimens from many locations, each with a unique set of resources. Producing successful outcomes in such varying environments requires an understanding of the fundamental pathophysiology of the disease, innovation in the application of basic treatment principles, and adjustment of outcome expectations to seek the best possible functional result using the resources available. These resources will change as socioeconomic conditions change and will drive changes in treatment protocols. An understanding of the pathophysiology and basic treatment principles should drive innovation in these changing times. PMID- 10829215 TI - Appliances and surgery for poliomyelitis in developing countries. AB - Death before maturity is the usual fate of the untreated crawling crippled child in developing countries. Most children with poliomyelitis, however, when upright and walking with supports or following surgery, are accepted by the community, educated by parents and relatives, and employable when they reach maturity. It is more economical to prevent 100 polio cases than to treat one hopelessly crippled child. It is often quicker to straighten 100 deformed limbs by simple subcutaneous surgeries than to treat a single patient with complicated procedures. It costs less for 100 crawling paralyzed children to walk in simple, locally made braces and clogs, than for 1 patient to be mobile in expensive imported appliances and boots. It is essential to educate or rehabilitate patients in addition to making them mobile. The final aim should be patients returned to their own village or town, accepted and integrated into their own communities, and earning their own living among their own friends. This chapter is discussed in further detail in both English and French in my Website, . This site also includes comprehensive information on both the manufacture of the simple appliances described and various surgical details. This information is also available on CD-ROM. PMID- 10829216 TI - Emerging multiresistant strains: recommended precautions in the emergency room and surgical setting. AB - The current success in treatment of surgical site infections may be jeopardized by the continued emergence of antibiotic resistance in bacteria common to these infections. The effectiveness of vancomycin against methicillin-resistant staphylococci may decrease as more cases of VISA emerge. No currently available antimicrobial is consistently effective against certain strains of VRE and the potential emergence of VRSA. Orthopaedic surgeons soon may be in the undesirable position of having to eradicate organisms resistant to all available antibiotics. Several new antibiotics show promising activity and may be useful against these multidrug-resistant bacteria. However, as the history of bacterial resistance has taught us, it likely only will be a matter of time until these organisms adapt mechanisms of resistance to these new drugs. The key then lies, as it always has, in preventive measures. Surgeons, and all physicians, must adhere to the precautionary guidelines recently set forth by the CDC and HICPAC. Chief among these guidelines is the elimination of inappropriate antibiotic usage, especially inappropriate vancomycin use. PMID- 10829217 TI - Appropriate screening for prevention of infection transmission by musculoskeletal allografts. PMID- 10829218 TI - Current issues and update on human immunodeficiency virus infection in the orthopaedic setting. AB - The risk of HIV transmission in the health care setting is a composite of risks, with exposure to blood being the most important source of infection and percutaneous injuries posing the greatest risk of transmission. The frequency of blood contact varies by occupation, procedure performed, and precautions used. Data suggest that PEP may be effective in reducing the risk of occupational HIV transmission. The risk for transmission of HIV from HCW to patients is very small. PMID- 10829219 TI - Issues and update on HIV infection in orthopaedic surgery: risk of surgery on HIV infected patients. PMID- 10829220 TI - Ethical considerations in orthopaedic surgery. AB - Because our actions as physicians have far-reaching consequences, and because society allows us to do things to others that no one else is free to do, physicians' professional activities fall under the domain of ethical evaluation. We are charged with the obligation to use specialized scientific knowledge, to work in concert with others, and to act for the good of our patients. In fact, acting for the good of our patients is the central tenet of ethical medical behavior. What constitutes the good of the patient, however, is not always clear. In general, we act to limit disease, restore function, alleviate suffering, and prolong life. We understand fully, however, that these goals may conflict with one another. Judgment about what is right for a particular patient leads us to another crucial consideration of ethical behavior, namely, respect for patient autonomy. We recognize that individuals have the right to control their own destiny. Patients have a right, therefore, to make choices about their medical care. As physicians, we must respect those rights. As such, certain ethical behavior is expected of us. We must be honest with our patients. We must provide them with accurate information on which to base their decisions. We must convey to them information about their diagnosis, prognosis, and treatment, even when it is unpleasant to do so. We must be open about our expertise and level of training for a particular procedure. We must respect their privacy and their right to withhold information even from family and friends. In short, we must respect their choices, even if we may disagree with those choices. To truly respect patient autonomy is to understand that, ultimately, the final decision lies with the patient. PMID- 10829221 TI - Teaching clinical ethics to orthopaedic surgery residents. PMID- 10829222 TI - Development of orthopaedic critical pathways. PMID- 10829223 TI - Mass spectrometric analysis of proteins. PMID- 10829224 TI - Protein sequence databases. PMID- 10829225 TI - Amino acid substitution matrices. PMID- 10829226 TI - Amino acid-based phylogeny and alignment. PMID- 10829227 TI - Searching databases to find protein domain organization. PMID- 10829228 TI - Identifying nature's protein Lego set. PMID- 10829229 TI - Evolution of domain families. PMID- 10829230 TI - Protein fold recognition using sequence profiles and its application in structural genomics. PMID- 10829231 TI - Protein sorting signals and prediction of subcellular localization. PMID- 10829232 TI - Gene and context: integrative approaches to genome analysis. PMID- 10829233 TI - Pathway databases and higher order function. PMID- 10829234 TI - Individual variation in protein-coding sequences of human genome. PMID- 10829235 TI - [Morphology and phenomenology of death by kicking (II)]. AB - In the years from 1982 to 1995, 20,504 autopsy cases were examined at the Departments of Forensic Medicine in Hamburg and Greifswald. In 103 cases death was caused by kicking. In the Greifswald area, the occurrence of kicking as the cause of death could be demonstrated in 68 cases (30% of all homicides), which is in contrast to the observations in Hamburg, were kicking amounted to only 2.5% of the homicides. Most of the victims (average age 44 years) were under the influence of alcohol (mean blood alcohol concentration 1.75/1000), and most were on social support. The victims often showed other signs of blunt trauma such as blows with the fist and strangulation. In a few cases, cutting and stabbing wounds were also found. In the majority of cases death was due to severe haemorrhages and aspiration of blood in addition to the direct effects of the head injuries. The crimes were committed by one person in 46 cases, two in 20 and three in 4 cases. The perpetrators (average age 27.6 years) were always younger than the victims. Case reports show that fatal kicking was mostly triggered by relatively trivial arguments. None of the incidents were preplanned by the perpetrators. In contrast to an expected higher incidence of kicking deaths in the urban area of Hamburg, these crimes occurred ten times more frequently in the rural area of Greifswald. PMID- 10829236 TI - [Cadaver dismemberment by chain saw]. AB - It is reported about the dismembering of a 57-years-old man by means of a chain saw, allegedly without participation in the very assault. The victim died after heavy violence to this throat and bleeding from stab wounds. Afterwards his trunk was divided horizontally, his thorax, skull and upper arm were sewn into without cutting through. The closer circumstances of the crime and localization of the saw injuries lead to the assumption of a participation in the assault. Technical details of chain saws and typical findings as to injuries are described. PMID- 10829237 TI - [Site, number and depth of wounds of the scalp in falls and blows--a contribution to the validity of the so-called hat brim rule]. AB - The sites, numbers and lengths of wounds of the skin of the scalp and head caused by blunt injuries (falls from standing position, falls downstairs, blows) in an autopsy series. Analysis of the localization, length and number of wounds located at the head in 305 autopsy cases of falls from a standing position (203 cases), falls downstairs (51 cases) and blows with relatively long materials (51 cases). For the distinction between falls and blows, among several other aspects the so called rule of the hat-brim-line is described: in cases of falls from a standing position down to a flat bottom, contusions can be expected in or near a line which represents the greatest horizontal circumference of the skull, whereas in cases of blows the contusions lay above this area. Although such a tendency was found in our material, a rule could not be confirmed: only approximately 55% of the wound due to blows were above this line, and in cases of falls from a standing position ca. 1/3 of the wounds were above this area. Only in the (dorso frontal and parietal) region "top of the head" contusions due to falls from a standing position were quite uncommon. The mean length and length distribution of wounds in the hat-brim-zone was not different between the 3 causes of injuries; in positions above this, wounds due to blows were longer compared to those by falls. In falls from a standing position, only exceptionally more than 1 wound was found, whereas in cases of homicide several or many wounds were common. Single wounds due to blows were only found in cases, in which the victims were knocked down but not killed by blunt forces, and death was caused by other methods (e.g. strangulation or stabbing). PMID- 10829238 TI - [Differentiation between self-inflicted and homicidal stab wounds to the neck]. AB - 39 autopsy cases with throat incision examined in the Bonn Institute of Legal Medicine in the years 1990 through 1998 were evaluated with respect to the query whether the morphological criteria generally established for the differentiation between homicide and suicide are also suitable for the assessment in this special localization. Integrative consideration of localization, direction of incision, hesitative resp. tentative injuries, defence injuries, concomitant injuries and capability to act allowed for a discrimination on grounds of legal medical findings in 84 per cent of the cases. PMID- 10829239 TI - [A gun silencer of a special kind]. AB - The authors report about a small bore pistol with silencer. In addition, the silencer can be sealed up by a rubber plate. This leads to a false estimation of the shooting distance. Also at close range shots the shot with the silencer and the rubber seal leaves hardly gunshot residues. The bullet wipe persists but microradiography shows his changed morphological composition. The use of the rubber seal leads predominantly to atypical bullet holes. PMID- 10829240 TI - [Forensic medicine experiences in Kosovo]. AB - In summer 1999, a German forensic team of CID officers and forensic pathologists was sent to Kosovo on request of the International Criminal Tribunal for the former Yugoslavia (ICTY) to investigate possible war crimes. For this purpose, witnesses had to be found and interrogated and graves of victims had to be located and the bodies exhumed and examined forensically. Roughly 200 bodies have been found and examined during the campaign that were predominantly of male sex and showed mainly gunshot wounds. A high percentage of all bodies could be identified. The article deals with different forensic-pathologic aspects of such an investigation. PMID- 10829241 TI - Atherogenetically relevant cells support continuous growth of Chlamydia pneumoniae. AB - The obligate intracellular bacterium Chlamydia pneumoniae has been implicated in the pathogenesis of atherosclerosis since viable pathogen has been recovered from plaques. Chlamydiae are epithelial pathogens notorious for causing persistent infection. Atherosclerosis, however, is a chronic inflammatory disease involving mesenchymal cells of the vascular wall. A bacterial contribution to atherosclerosis appears more relevant if the resident mesenchymal cells of the vascular wall that constitute the plaque can support chlamydial infection continuously. Therefore we inoculated immortalized and primary mesenchymal cells with a vascular and a respiratory Chlamydia pneumoniae isolate. Primary human coronary artery endothelial and smooth muscle cells, primary human embryonic fibroblasts as well as the immortalized cell lines were permissive for continuous growth of both strains. Thus, the resident vascular cells that produce the atheromatous plaque can acquire permanent productive. Chlamydia pneumoniae infection. Immortalized monocytic cells and peripheral blood monocytes also supported chlamydial growth, though productive infection ceased after 5 passages. Monocytes/macrophages are not resident cells of the vascular wall but have an active role in plaque formation. Systemic circulation and transendothelial migration makes them a potential vector system for chlamydial distribution. These findings add further plausibility to the hypothesis of a chronic infectious component in the multifactorial condition of atherosclerosis. Further studies must precisely define chlamydial target cells in vivo and differentiate infection in resident cells of the vascular wall from a presence limited to migrating macrophages. Endovascular infection might provide an explanation for unclear phenomena of atherogenesis like mesenchymal cell proliferation and its distinct inflammatory component. PMID- 10829242 TI - Immune responses to Chlamydia antigens in atherosclerosis. AB - Aim of this study was to isolate T lymphocytes from atheromatous plaques and to determine they respond to Chlamydia antigens. Atheromatous plaques from carotid endarterectomy patients, were cultured in vitro with the T cell growth factor, IL 2. This rarely allowed outgrowth of T cell lines. However, when combined with a mitogenic or antigenic stimulus to T cells, T cell lines were obtained from most patients, and from approximately 30% of replicate plaque tissue fragments. Chlamydia organisms were as effective in allowing the establishment of T cell lines as other recall antigens. T cell lines were tested for their ability to recognize antigens presented by autologous macrophages. Some lines responded to Chlamydia organisms, and also to the recombinant Chlamydia proteins hsp60 and OMP2. However, other lines recognized recall antigens. These results indicate that the atheromatous plaque contains memory T lymphocytes, and amongst the antigens they recognize are Chlamydia proteins. Stimulation of T cells was required to allow outgrowth in vitro, suggesting that the T cells were not in an activated state in vivo. However, since Chlamydia pneumoniae is present in the atheromatous plaque, activation of Chlamydia-reactive T cells by local antigen is a potential pro-inflammatory mechanism which could contribute to the pathogenesis of atherosclerosis. PMID- 10829243 TI - Chlamydia pneumoniae infection is not an independent risk factor for arterial disease. AB - Appreciation of atherosclerosis as an infectious disease has fostered interest in the role that Chlamydia pneumoniae may play in atheroma development. Although data from seroepidemiological and experimental studies have established an association between the pathogen and atherosclerosis, little is known about how the organism contributes to lesion development. Atherosclerosis is a complex disease process and the role of any pathogen must be considered in the context of other risk factors. Here we focus on the relationship between Chlamydia pneumoniae and conventional risk factors for atherosclerosis. There is evidence for a strong association between chronic infection with Chlamydia pneumoniae and smoking as well as high serum cholesterol. It is concluded from the present data that chronic infection with the pathogen is not an independent risk factor for atherosclerosis. PMID- 10829244 TI - Pathogenesis of atherosclerosis: infectious versus immune pathogenesis. A new concept. AB - A novel concept for the pathogenesis of atherosclerosis is presented that obviates the need to search for a causative infectious agent. It is shown that LDL stranded in the subendothelium is transformed by enzymatic (non-oxidative) modification to yield an entity with complement- and macrophage-activating properties. This primarily serves to clear the stranded lipoprotein from tissues. However, sustained overactivation of complement and macrophages occurs when LDL deposition is too extensive. A chronic inflammation then ensuing initiates and promotes the pathology of the atherosclerotic lesion. Atherosclerosis is thus a special form of an immunopathological disease which evolves through overactivation of the most archaic effector components of the immune system by altered self. PMID- 10829245 TI - Atherosclerosis--an autoimmune disease due to an immune reaction against heat shock protein 60. AB - This brief overview, our concept of an autoimmune pathogenesis of atherosclerosis is summarized. In principle, we postulate that we may have to "pay" for our protective immunity against microbial heat-shock protein 60 (hsp60) with the danger of a cross-reactivity with our own hsp60 that is expressed by endothelial cells that are stressed by classical risk factors for atherogenesis such as hypertension, high serum cholesterol levels, components of cigarette smoke and other toxins, etc. hsp60 are phylogenetically highly conserved and there is an over 55% homology between bacterial hsp60 and the human homologue forming the basis for this cross-reactivity. As another possibility, the initiation of the disease by a bona fide autoimmune reaction against chemically altered autologous hsp60 is discussed. PMID- 10829246 TI - Restenosis after percutaneous coronary interventions and infection. AB - Infectious agents may directly or indirectly (through the response of the host's immune system) modulate the growth of vascular cells. Local and/or systemic increase of cytokines could influence the extent of (re-)stenosis in the vascular tree. Further studies in this field may identify patients at a high risk for atherogenesis and restenosis. Their results should be helpful in treating restenosis after percutaneous coronary interventions. PMID- 10829247 TI - Transplant vasculopathy: a model for coronary artery disease? AB - Although transplant vasculopathy and native atherosclerosis are clinically and pathologically different entities, the pathogenesis of both diseases exhibits some common mechanisms. Both may be regarded as responses to injury within a broadened concept of the immune system. Alloantigens (e.g. on donor endothelial cells) or autoantigens (e.g. oxydized LDL cholesterol) are presented by antigen presenting cells to the T cells of the body's immune system. With the appropriate costimulatory signal, this signal pattern generates a differentiated T cell, B cell, and inflammatory cell response whereas without the second signal, the immune cells undergo apoptosis. In case of immune cell proliferation and differentiation, a coordinated pattern of cytokine release is initiated. Monocyte derived macrophages are also involved in this process which culminates in rolling, sticking, and diapedesis through the coronary vascular endothelium and phenotype switch of medial smooth muscle cells mediated by generation of growth promoting cytokines. Thus, viewed within a broadened paradigm of the immune system's role both disease entities may represent different vignettes of an integrated pathophysiological response to an endothelial injury. PMID- 10829248 TI - Cell cycle proteins and atherosclerosis. AB - Progression of the eukaryotic cell through the cell cycle to induce cell proliferation is a fundamental event in developmental growth processes. Specific cell cycle proteins are critical for either inducing or suppressing the cell cycle. These proteins, therefore, have been found to be key players in regulating cell proliferation in diseases like cancer and atherosclerosis. The present manuscript reviews the process of cell proliferation in atherosclerosis and the data that have implicated the various cell cycle proteins in restenotic and atherosclerotic vascular disease. PMID- 10829249 TI - Inflammation and acute coronary syndromes. AB - The presence of inflammatory infiltrates in unstable coronary plaques suggests that inflammatory processes may contribute to the pathogenesis of these syndromes. In patients with unstable angina, coronary atherosclerotic plaques are characterized by the presence of macrophages, and to a lesser extent, T lymphocytes, at the immediate site of either plaque rupture or superficial erosion; moreover, the rupture-related inflammatory cells are activated, indicating ongoing inflammation at the site of plaque disruption. These observations are confirmed by clinical studies demonstrating activated circulating neutrophils, lymphocytes and monocytes, and increased concentrations of pro-inflammatory cytokines, such as interleukin (IL) 1 and 6, and of acute phase reactants in patients with unstable angina and myocardial infarction. In particular elevated levels of C-reactive protein are associated with an increased risk of in-hospital and 1 to 2 years new coronary events in patients with unstable angina, but are also associated with an increased long-term risk of death and myocardial infarction in apparently normal subjects. Thus, accumulating evidence suggests that inflammation may cause local endothelial activation and, possibly, plaque fissure, leading to unstable angina and infarction. Although no information is yet available on the causes of inflammation and on its localization, these novel lines of research may open the way to a different approach to the patient with acute coronary syndromes. PMID- 10829250 TI - Strategies to optimize CAD prevention in modern cardiology. The "Marburg CAD Prevention Project". AB - In recent years we identified numerous cardiovascular risk factors and had been able to reduce cardiovascular events by a variety of different interventions. There is no question that we can improve the course of coronary artery disease (CAD) in patients with a high-risk profile by modifying these factors. Despite this knowledge, many patients with known CAD or myocardial infarction are not treated for secondary prevention as recommended by well established guidelines (http:?www.chd-taskforce.com). In order to improve secondary prevention in our patients we started a project, the so called "Marburg CAD Prevention Project". By this we combine our computer data base of the CAD preventional routine laboratory with the computer program CARDDAS. Individual risk factors and the angiographic findings were analyzed. Patients as well as local physicians were informed on the need to treat the important risk factors. This approach ensures that at least all of our patients with documented CAD receive the appropriate preventional recommendations and treatment. PMID- 10829251 TI - HMG-CoA reductase inhibition: anti-inflammatory effects beyond lipid lowering? AB - Atherosclerosis has many features of a chronic inflammatory disease. Atherosclerotic lesions contain inflammatory cells like activated T-lymphocytes and macrophages. Systemic markers of inflammation such as white blood cells, C reactive protein, serum amyloid A, interleukin 6 and soluble adhesion molecules are predictive of future cardiovascular events, even after adjustment for the contribution of established cardiovascular risk factors. Atherogenic lipoprotein particles, in particular modified low-density lipoproteins (LDL), elicit pro inflammatory responses of cellular elements of the vessel wall, including endothelial dysfunction and activation of monocyte-derived macrophages. Treatment, with HMG-CoA reductase inhibitors has proven the most successful strategy to reduce the concentration of LDL in the circulatory system. These compounds lower LDL cholesterol by inhibiting the mevalonate pathway in the liver, which in turn depletes the regulatory pool of cholesterol and enhances the activity of LDL receptors. Five prospective clinical trials have convincingly demonstrated that HMG-CoA reductase inhibitors can effectively lower the incidence of cardiovascular events in primary and secondary prevention. Post hoc analyses of these trials suggest that the clinical benefit brought about by HMG CoA reductase inhibitors may not entirely be due to their effect on the levels of circulating lipoproteins. In-vitro observations of anti-inflammatory actions of HMG-CoA reductase inhibitors on vascular cells have been suggested to explain effects beyond lipid-lowering. It is, however, not clear whether these findings are relevant to the in-vivo situation. Further investigation is now necessary in order to determine the relative significance of cholesterol lowering and of ancillary effects to the overall clinical benefit of statin treatment. PMID- 10829252 TI - The VEGF splice variants: properties, receptors, and usage for the treatment of ischemic diseases. AB - Vascular endothelial growth factor (VEGF) was discovered 10 years ago as a growth factor that can regulate angiogenesis and in addition the permeability of blood vessels. Numerous studies have revealed that it is essential for normal embryonic development and that it plays a major role in physiological and pathological events of angiogenesis in adults. It is unique in that its expression is regulated directly by hypoxia. These properties are now being exploited in attempts aimed at the induction of new blood vessels in pathological situations such as ischemic heart disease. Five VEGF forms of 121 to 206 aminoacids are produced from a single gene by alternative splicing. Cells expressing VEGF usually express several forms simultaneously. VEGF121 does not contain exons 6 and 7 of the gene and consequently lacks a heparin binding ability. However, this form is fully active as an inducer of angiogenesis, and as a blood vessel permeabilizing agent. Exon 6 and 7 contain 2 independent heparin binding domains. The VEGF form containing exon 7 (VEGF165) and the vascular endothelial growth factor form containing exon 6 (VEGF145) display similar biological potencies raising the question of why so many VEGF forms are required. It was found that VEGF121 diffuses better because it does not bind to heparan-sulfate proteoglycans. In contrast, VEGF145 binds to extracellular matrix and is released from it slowly. When the receptor binding properties of VEGF121 and VEGF165 were compared it was found that VEGF165 binds to a class of VEGF receptors that is not recognized by VEGF121. These receptors are encoded by the neuropilin-1 gene, and we have recently found that the related neuropilin-2 gene also encodes a VEGF165 receptor. We have recently found evidence indicating the neuropilins form complexes with another VEGF receptor, VEGFR-1. However, the biological function of this complex remains to be elucidated. PMID- 10829253 TI - Potassium channel openers and blockers in coronary artery disease. Comparison to betablockers and calcium antagonists. AB - Potassium channel openers and blockers, which belong to a novel class of vasodilator drugs and to the class of specific bradycardic substances, are potential new antianginal drugs. Experimental findings in vivo suggest that bimakalim is a new substance characterized as ATP-sensitive K+ channel openers, since it exerts preferential vasodilation of the collateral circulation of the coronary vasculature and both leads to increase blood flow to ischemic areas and to attenuate the ST segment elevation caused by regional ischemia in the canine heart. Opening of KATP increases the conductivity of potassium ions which results in hyperpolarization of smooth muscle membranes, thus producing vasodilation. Tedisamil is a new bradycardic agent proven to exert antiischemic and antiarrhythmic effects by blockade of the cellular cardiac repolarization K+ currents as well as of multiple neuronal and vascular K+ currents (Ito, Ik, and K+ATP). Using right heart catheterization and exercise tolerance tests, we investigated the hemodynamic, antiischemic and neurohumoral effects of bimakalim and tedisamil in patients with angiographically proven coronary artery disease, stable angina pectoris and reproducible ST segment depression during exercise. In 50 patients with coronary artery disease, the hemodynamic and antiischemic effects of a single oral dose bimakalim of 0.1 mg, 0.3 mg and 0.6 mg were compared to placebo. In a dose-finding baseline-controlled study, a comparable collective was examined for the effects of acute i.v. administration of tedisamil 0.1, 0.2, 0.3 and 0.4 mg/kg bw. A subgroup of 8 patients receiving 0.3 mg/kg bw tedisamil i.v. was compared with a similar group of 14 patients who had received esmolol (i.v. bolus of 500 micrograms/kg, maintenance dose 200 micrograms/kg/min) and gallopamil (initial dose 0.025 mg/kg, maintenance dose 0.0005 mg/kg/h) in a second intra-individual comparison. Furthermore, in 48 patients, short-term (6 days) effects of tedisamil, 2 times 100 mg orally, were compared to 2 times 50 mg atenolol treatment. With a single oral dose of bimakalim antianginal and/or antiischemic effects were lacking, increased doses, however, induced changes in hemodynamics typical of vasodilation, i.e., a significant decrease in systolic blood pressure and a secondary chronotropic response. In contrast to bimakalim, tedisamil produced antiischemic effects and was found to have favorable hemodynamic, neurohumoral and antiischemic effects in comparison to the betablocker esmolol and atenolol in patients with coronary artery disease. Tedisamil induced a dose-dependent decrease in both heart rate and the index of myocardial oxygen consumption associated with an improvement in ST segment depression. Tedisamil as well as esmolol and atenolol showed almost equipotent antiischemic effects at the doses administered. Compared with gallopamil, both tedisamil and esmolol were superior in their effects on myocardial oxygen consumption and ST segment depression, whereas plasma lactate concentrations were more reduced by tedisamil and gallopamil. PMID- 10829254 TI - [Second and Third Annual Report of the German Society of Coronary Angiography and Interventions in Private Practice (BNK) concerning quality assurance in invasive cardiology]. AB - Since 1995, the German Society for Cardiac Angiography and Interventions in Private Practice (BNK) has been intensely involved in quality assurance; since 1996, it has registered relevant data for diagnostic left heart catheterization (DIAG) and coronary interventions (PTCA). The following data are based on a total of 72,777 cardiac catheters, of which 54,513 were DIAG and 18,264 were PTCA (DIAG in 1996: 10,316; in 1997: 15,692 and in 1998: 28,505. PTCA in 1996: 2,597; in 1997: 5,600 and in 1998: 10,067). The mean patient age was 61.9 years for males and 65.3 years for females (31%). DIAG was performed on an out-patient basis in 60% of the cases. The proportion of self-referrals is circa one-third and has not increased over time. A s/p PTCA was present in 21% of the DIAG patients; those after coronary bypass surgery showed a steadily increasing trend towards a follow up DIAG (8% of the patients). There was a distinctive decrease in the use of contrast medium from an average of 164 ml (1996) to 138 ml (1998). The symptoms leading to DIAG did not substantially change in the years from 1996 to 1998; the angiographic range of the coronary disease and the degree of an impaired myocardial function have remained virtually unchanged. The treatment recommendations appear to tend towards medical therapy with decreasing indications for PTCA (20%) and open heart surgery (16%). The mean duration of a PTCA procedure decreased from 57 +/- 8 (1996) to 44.6 +/- 25 (1998) minutes. The success rates remained unchanged for coronary stenoses as well as for recanalization of chronic total occlusions. PTCA complications: the incidence of abrupt coronary occlusions was 2% and of emergency bypass surgery 0.4%; mortality was 0.1%. PMID- 10829255 TI - Principles of selection and use of antibacterial agents. In vitro activity and pharmacology. AB - The selection of an antimicrobial treatment regimen is based on many factors, including the nature of the infection, the identity and susceptibility pattern of the infecting organisms, and the pharmacokinetics and pharmacodynamics of the antibacterial drugs. This article discusses principles of susceptibility testing, pharmacology, and monitoring of therapy. PMID- 10829256 TI - Pharmacodynamics of antibacterial drugs. AB - Pharmacodynamics of antibacterial agents relates the time course of drug concentration to its antimicrobial effects at the infection site. Antibacterial agents can be divided into three groups based on pharmacodynamic characteristics: agents that exhibit concentration-dependent bactericidal activity over a range of drug concentrations (e.g., aminoglycosides and fluoroquinolones); agents that exhibit time-dependent bactericidal activity that has little relationship to the magnitude of concentration, provided the concentrations are above a minimally effective level (e.g., beta-lactam antibiotics and vancomycin); and agents that exhibit a predominantly bacteriostatic effect. Knowledge of antimicrobial pharmacodynamics provides a rational basis for determining optimal regimens of dosage amounts and length of dosage intervals. PMID- 10829258 TI - Topical antibacterial agents. AB - Topical antibacterial agents occupy an important niche of antimicrobial therapy for both inpatients and outpatients. These agents, including antiseptic and antibiotic preparations, are used for prophylaxis and treatment of infection. Prophylactic uses include application for traumatic and surgical wounds, burns, intravascular catheters, and eradication of S. aureus nasal carriage. Topical antibacterial agents are also used for treatment of primary and secondary pyodermas. Individual antibacterial agents have been reviewed. Of note, despite the widespread use of topical antibacterial agents, further data on which to guide therapy are needed in many instances. PMID- 10829257 TI - Pathogens resistant to antimicrobial agents. Epidemiology, molecular mechanisms, and clinical management. AB - The emergence of resistance to antimicrobial agents continues to be a major problem in the nosocomial setting and now in nursing homes and the community as well. Bacteria use a variety of strategies to avoid the inhibitory effects of antibiotic agents and have evolved highly efficient means for the dissemination of resistance traits. Control of antibiotic-resistant pathogens provides a major challenge for both the medical community and society in general. To control the emergence of resistant pathogens, CDC and infection control guidelines must be adhered to, and antibiotics must be used more judiciously. PMID- 10829259 TI - Antibacterial agents in pediatrics. AB - From low birth weight infants to adolescents, physiologic and developmental differences underlie the marked differences in pharmacokinetics and pharmacodynamics of antibacterial agents. Certain diseases, such as cystic fibrosis, also can alter these parameters. This article describes the principles of pharmacokinetics and pharmacodynamics that are unique to children and that characterize the clinical application of selected antibacterial agents to infectious diseases in children. PMID- 10829260 TI - Antibiotic agents in the elderly. AB - Changes that occur in the pharmacology of drugs in the elderly must be considered in the use of antimicrobial agents. Although absorption of orally administered drugs is not affected in a significant way, renal function decreases, drug-drug interactions increase, compliance with regimens may be decreased, and drug toxicity is increased. The most frequent infections occurring in the elderly are pneumonia, urinary tract infection, and soft-tissue infection. CDAD is usually a complication of antibiotic therapy. Pneumonia can be categorized as community acquired, LTCF, and hospital-acquired. Therapeutic approaches vary according to which of these sites is involved. Urinary tract infection is divided into upper tract infection, lower tract infection, and asymptomatic bacteriuria. Upper tract infection is treated for a longer period than lower tract infection; with few exceptions, asymptomatic bacteriuria is usually not treated. Soft-tissue infection is usually caused by an infected pressure ulcer or cellulitis (which may be a complication of a diabetic foot ulcer or an ulcer due to peripheral vascular disease). These infections have different microbial causes and require different therapeutic approaches. PMID- 10829261 TI - Use of antibacterial agents in renal failure. AB - This article provides information on the pharmacokinetics of antibacterial agents in patients with normal renal function and those with impaired renal function. Specific discussion includes the use of serum levels, dosage adjustments in dialysis, new strategies for cefazolin dosages in dialysis patients, and antibiotic toxicity in renal failure, and tabular data is presented for determining appropriate dosages for varying degrees of renal failure. PMID- 10829262 TI - Antibacterial agents in infections of the central nervous system. AB - Experimental animal models have provided information applicable to antimicrobial therapy of infections of the central nervous system. The efficacy of an antimicrobial agent in the therapy of bacterial meningitis depends on its ability to penetrate the blood-brain barrier, its activity in purulent cerebrospinal fluid, and a demonstration of rapid bactericidal activity against the offending pathogen. The recent emergence of resistant pathogens is challenging the therapy for bacterial meningitis. Various strategies for treating resistant pathogens have been evaluated in experimental animal models. Encouraging results have led to clinical trials to evaluate the efficacy of newer agents, alone or in combination with standard regimens. PMID- 10829263 TI - Ureidopenicillins and beta-lactam/beta-lactamase inhibitor combinations. AB - Although research and development of new penicillins have declined, penicillins continue to be essential antibiotics for the treatment and prophylaxis of infectious diseases. The most recent additions are the ureidopenicillins and beta lactam/beta-lactamase inhibitor combinations. This article reviews the spectrum of activity, toxicity, pharmacokinetics, and clinical uses of the ureidopenicillins, and the beta-lactam/beta-lactamase inhibitor combination agents. PMID- 10829264 TI - Cephalosporins, carbapenems, and monobactams. AB - Nonpenicillin beta-lactams exhibit a variable spectrum of antimicrobial activity, have a wide range of clinical uses and a favorable safety profile. Cefepime's twice-daily dosage and increased activity against Enterobacteriaceae may offer some advantages over older cephalosporins. The carbapenems offer a broad antimicrobial spectrum, and meropenem has an improved safety profile compared with imipenem. Aztreonam is a useful alternative for patients with aerobic gram negative infections who are allergic to penicillin. The emergence of resistant organisms, however, is an increasing problem with the frequent use of these antibiotics. PMID- 10829265 TI - The newer macrolides: azithromycin and clarithromycin. AB - Azithromycin and clarithromycin are two relatively new macrolide antimicrobial agents. Although azithromycin and clarithromycin are structural analogues of erythromycin, they offer distinct advantages in comparison. This article reviews the pharmacokinetics, antimicrobial activity, clinical use, and adverse affects of these antimicrobial agents. PMID- 10829266 TI - Antibiotics for gram-positive bacterial infections. Vancomycin, teicoplanin, quinupristin/dalfopristin, and linezolid. AB - Vancomycin is a safe, effective antibiotic for a variety of serious gram-positive infections. Because of emerging resistance in enterococci and staphylococci and the emerging threat of spread of vancomycin-resistant genes to other gram positive organisms, judicious use of vancomycin should be promoted. Quinupristin/dalfopristin, a streptogramin antibiotic, and linezolid, an oxazolidinone, show promise against some strains of gram-positive bacteria that are resistant to vancomycin. PMID- 10829267 TI - Once-daily dosing of aminoglycoside antibiotics. AB - Improved understanding of the pharmacodynamics and toxicity of aminoglycoside antibiotics has resulted in the study of once-daily dosing regimens. Although studies have suggested a therapeutic advantage and possibly a decrease in toxicity with once-daily administration, these effects have been modest. The cost savings associated with once-daily aminoglycoside administration, however, makes this approach appealing. Although a syndrome of fever, tachycardia, hypotension, and rigors has been associated with once-daily dosing of gentamicin, this appears to have been the result of impurities in the antibiotic from a single offshore supplier. This syndrome has not been associated with other aminoglycoside antibiotics, and the FDA has now withdrawn its recommendation that once-daily aminoglycoside use be avoided. As with any medical regimen, the decision to use once-daily dosing of aminoglycoside agents must take into account special patient characteristics and the disease state being treated. Although once-daily dosing appears effective in limited studies in children, in individuals with neutropenia, and in individuals with cystic fibrosis, its role in gram-positive coccal endocarditis and in individuals with altered volumes of distribution remains uncertain. Further data are needed to clarify the role of once-daily dosing in these situations. PMID- 10829268 TI - Fluoroquinolones. AB - The fluoroquinolone class of antimicrobial agents has expanded dramatically in the last 5 years and will continue to grow over the next decade. This article discusses the newer fluoroquinolones in detail, including pharmacokinetics, pharmacodynamics, safety, and drug interactions, and the spectrum of in vitro activity. Newer agents are compared and contrasted with the older ones, particularly ciprofloxacin and ofloxacin, and problems with liver toxicity and trovafloxacin are described. Finally, appropriate use of the fluoroquinolones is discussed, including their role in the treatment of urinary tract infections, sexually transmitted diseases, gastrointestinal infections, osteomyelitis, and respiratory tract infections. PMID- 10829269 TI - Advanced 5'-silyl-2'-orthoester approach to RNA oligonucleotide synthesis. PMID- 10829270 TI - Preparation of specifically 2H- and 13C-labeled ribonucleotides. PMID- 10829271 TI - Base-modified phosphoramidite analogs of pyrimidine ribonucleosides for RNA structure-activity studies. PMID- 10829272 TI - Use of T7 RNA polymerase and its mutants for incorporation of nucleoside analogs into RNA. PMID- 10829273 TI - Phosphorothioate modification of RNA for stereochemical and interference analyses. PMID- 10829274 TI - Chemical probing of RNA by nucleotide analog interference mapping. PMID- 10829275 TI - Joining of RNAs by splinted ligation. PMID- 10829276 TI - Heavy atom derivatives of RNA. PMID- 10829277 TI - Site-specific cleavage of transcript RNA. PMID- 10829278 TI - Using DNAzymes to cut, process, and map RNA molecules for structural studies or modification. PMID- 10829279 TI - Purification, crystallization, and X-ray diffraction analysis of small ribozymes. PMID- 10829280 TI - Solving large RNA structures by X-ray crystallography. PMID- 10829281 TI - Conventional and time-resolved ribozyme X-ray crystallography. PMID- 10829282 TI - Nuclear magnetic resonance methods to study RNA-protein complexes. PMID- 10829283 TI - Filamentous bacteriophage for aligning RNA, DNA, and proteins for measurement of nuclear magnetic resonance dipolar coupling interactions. PMID- 10829284 TI - Biochemical and nuclear magnetic resonance studies of aminoglycoside-RNA complexes. PMID- 10829285 TI - Experimental prerequisites for determination of tRNA binding to ribosomes from Escherichia coli. PMID- 10829286 TI - Three-dimensional cryoelectron microscopy of ribosomes. PMID- 10829287 TI - Preparation of functional ribosomal complexes and effect of buffer conditions on tRNA positions observed by cryoelectron microscopy. PMID- 10829288 TI - Probing RNA structure and function, by circular permutation. PMID- 10829289 TI - Kinetic oligonucleotide hybridization for monitoring kinetic folding of large RNAs. PMID- 10829290 TI - Time-resolved synchrotron X-ray footprinting and its application to RNA folding. PMID- 10829291 TI - Analysis of global conformation of branched RNA species using electrophoresis and fluorescence. PMID- 10829292 TI - Application of circular dichroism to study RNA folding transitions. PMID- 10829293 TI - Fluorescence assays to study structure, dynamics, and function of RNA and RNA ligand complexes. PMID- 10829294 TI - Transient electric birefringence for determining global conformations of nonhelix elements and protein-induced bends in RNA. PMID- 10829295 TI - Structure-function relationships of RNA: a modeling approach. PMID- 10829296 TI - Computational modeling of structural experimental data. PMID- 10829297 TI - Modeling RNA tertiary structure from patterns of sequence variation. PMID- 10829298 TI - Hepatic haemodynamic changes following inhibition of endothelium-derived relaxing and hyperpolarising factors in anaesthetised miniature pigs. AB - The influence of endothelium-dependent vasodilatation in regulating the hepatic circulation has been investigated by intraportal infusion of inhibitors of either endothelium-derived relaxing factor (NG-nitro-L-arginine-methyl-ester [L-NAME]) or of endothelium-derived hyperpolarising factor (ATP-dependent K(+)-channel inhibitor, glybenclamide) in barbiturate anaesthetised miniature pigs. Intraportal infusion of acetylcholine (5.5 micrograms kg-1 min-1 over 2 min) produced a selective 3-fold increase in hepatic artery and coeliac trunk blood flow, while mesenteric, portal, systemic, and pulmonary vascular beds remained unchanged. Intraportal L-NAME or glybenclamide did not reduce the hepatic artery and coeliac trunk flows but increased systemic and mesenteric vascular resistances. The acetylcholine-induced hepatic artery vasodilatation was partially blocked by 59%, 76% and 66% by L-NAME, at 30, 100, and 300 mg/kg respectively. Glybenclamide pretreatment up to 3 mg/kg did not modify acetylcholine-induced vasodilatation of the hepatic artery and coeliac trunk. Furthermore, prior cyclooxygenase inhibition did not alter the hepatic vascular response to acetylcholine. These results suggest that, in contrast to what is observed in large vessels, the hepatic vascular tree may not be entirely regulated by nitric oxide under basal conditions, but nitric oxide is released readily upon stimulation with acetylcholine, a response that is largely but incompletely blocked by L-NAME pretreatment. Neither basal vascular tone nor acetylcholine-induced vasorelaxation are mediated by the opening of glybenclamide sensitive K+ channels in the hepatic circulation in pigs. PMID- 10829299 TI - Identification of genes differentially expressed in melanoma sublines derived from a single surgical specimen characterised by different sensitivity to cytotoxic T-lymphocyte activity. AB - In this study we used differential display technology in an attempt to obtain an insight into the mechanisms underlying escape of tumour cells to the specific cytotoxic T cell response. A primary tumour cell line and autologous tumour infiltrating lymphocytes were raised from a metastatic melanoma sample (ME15). Upon co-culture of tumour infiltrating lymphocytes with irradiated tumour cells, CTL specific for neoplastic cells were generated and cloned. Using a CTL clone, a cytotoxicity resistant tumour subline (ME15R) was immunoselected. We applied a PCR-based differential display technique to amplify DNA sequences differentially expressed in ME15 sublines sensitive (S) or resistant (R) to specific CTL killing. 10 different sequences whose expression was exclusively detectable in ME15S cells were identified. Five of them matched with known expressed sequence tags encoding products of unidentified function. 2 showed high homology with a mitochondrial mRNA and with the gene encoding the S24 ribosomal protein. Most interestingly, genes coding for glutamine synthetase, TGF-beta-3 and PAX3, a well characterised transcription factor, were only expressed in ME15S cells. The latter gene was found to be transcribed in all healthy tissues tested, but only in a subgroup of established melanoma cell lines. Taken together, our data underline the relevant potential of differential display technology in the molecular analysis of paired tumour lines endowed with different phenotypic characteristics. Cloning of entire open reading frames and transfection studies are warranted to clarify the role of individual differentially displayed genes in the escape of tumour cells from cellular immune response. PMID- 10829300 TI - [Bilateral adrenal hemorrhage with adrenal insufficiency in the framework of primary antiphospholipid antibody syndrome]. AB - We report a case of acute adrenal insufficiency in a context of probable bilateral adrenal haemorrhage, as revealed by CT-scan in a 52-year-old woman with a history of spontaneous abortion and repeated ischaemic stroke without symptoms or signs of collagen vascular disease. The symptoms began after the patient had successfully been treated for pneumonia. She had persistently high titres of IgG anticardiolipin antibodies, antibodies against beta 2-glycoprotein I and a lupus anticoagulant. The diagnosis of primary antiphospholipid syndrome with adrenal insufficiency was postulated. PMID- 10829301 TI - [What is the evidence with regard to the effectiveness of orlistat?]. AB - BACKGROUND: Obesity is worldwide one of today's most important medical and public health problems. Orlistat (Xenical) is a relatively new drug in the pharmacological treatment of obesity which partially blocks fat absorption. The following article analyses the available evidence of orlistat's effectiveness in the treatment of obese patients. METHODS: Three randomised controlled trials investigating the effect of orlistat in the treatment of obesity were identified by systematic Medline search. The internal and external validity of these studies was assessed using systematic criteria. RESULTS: All three studies consistently demonstrate a treatment benefit of orlistat compared to placebo. Patients treated with orlistat lost an average of 3.4 kg more than patients taking placebo while on a hypocaloric diet. Simultaneously, control of cardiovascular risk factors improved independently of the observed weight loss. Up to 40% of all patients experienced gastrointestinal side effects which were generally well tolerated. The studies prove that treatment with orlistat can result in a moderate weight reduction. However, the results of the studies cannot be easily generalised to obese patients in a primary care setting, due to limitations concerning the studies' internal and external validity. CONCLUSIONS: Based on these studies orlistat is an efficient pharmacological treatment for obesity in patients adhering to a hypocaloric diet. Studies demonstrating orlistat's effectiveness in a primary care setting are so far lacking. From a public health perspective there is a need for a randomised controlled trial showing not only orlistat's effectiveness on surrogate markers in a primary care setting but, ideally, a reduction in obesity-related mortality and morbidity. PMID- 10829302 TI - [Blue sclera and chest pain]. PMID- 10829303 TI - [Breast cancer]. PMID- 10829304 TI - [Epidemiology and prevention of breast cancer]. AB - Breast cancer is the leading cause of cancer in females in Austria (25% of all newly diagnosed cancer cases). So far no comprehensive theory about etiology exists. Risk factors generally considered to be established are: demographic, reproductive and hormonal factors, diet, benign breast disease, familial aggregation and genetics. Most of the risk factors cannot be affected by primary prevention. Nevertheless women should be informed about cancer risk and lifestyle factors. At present secondary prevention through mammographic screening can reduce the impact of breast cancer morbidity and mortality. In Austria the reduction of advanced cancers at diagnosis, followed by a reduction of mortality rates is plausibly related to screening prevalence. PMID- 10829305 TI - [Breast cancer from the viewpoint of the pathologist]. AB - Important criteria for histopathological evaluation in breast carcinoma are described. Of major importance is macroscopical evaluation. It is mainly the evaluation of the resection margins which still remains a clinically relevant problem for pathologists. New trends for intraductal carcinoma and evaluation of axillary lymph nodes are reported. An overview for prognostic factors is given. PMID- 10829306 TI - [Surgical and plastic reconstructive therapy of breast carcinoma]. AB - Breast conserving surgery is the therapy of choice in breast cancer patients and may avoid mastectomy in most of the patients. New treatment options such as sentinel lymph node biopsy may avoid complete axillary dissection in selective patients. Plastic reconstructive procedures like immediate reconstruction or oncoplastic surgery decrease physical sequelae and increase the quality of life and should be offered to all breast cancer patients: Modern breast cancer management requires intense cooperation of all cancer subdisciplines. PMID- 10829308 TI - [Multimodal tumor therapy--evidence for effectiveness?]. PMID- 10829307 TI - [Drug therapy of breast carcinoma]. AB - Progress in the systemic therapy of breast cancer, especially in the adjuvant situation, based on systematically performed clinical trials, represents a paradigm for applied clinical research in general and in particular for oncology. This survey will present detailed indications for the evidence-based use of chemotherapy and hormone therapy in breast cancer, based on the relevant prognostic and predictive factors available. Drugs, both conventional and still under investigation, as well as therapeutic strategies will be reviewed with respect to their use in the adjuvant and the palliative situation. Explicit distinction will be made between standard therapies on the one side and expected new developments on the other side. Therapeutic details, general therapeutic guidelines as well as critical appraisals of recent developments such as high dose chemotherapy or chemoprevention will be considered. PMID- 10829309 TI - [Biological principles for multimodal therapy approaches]. AB - Cancer development and progression has been associated with numerous genetic events in tumor cells. Germline mutations of caretaker and gatekeeper genes are responsible for hereditary cancer syndromes. Exogenous factors in conjunction with functional germline variants of a variety of genes may contribute to tumor initiation in sporadic malignant disease. Furthermore, pathways to neoplasia require somatic events in the developing tumor. Acquired or inherited genetic instability permits stepwise tumor progression. The most fearsome aspect of tumor progression is dissemination of tumor cells to draining lymph nodes of the primary or to distant organs, which limits effectiveness of surgical therapy. Cellular heterogeneity of malignant neoplasms has important implications for chemotherapy and radiotherapy. An increasing understanding of the molecular biology of tumors is the prerequisite for improved prediction, prevention and therapy of malignant disease. PMID- 10829310 TI - [What is the value of neoadjuvant therapy in bronchial carcinoma?]. AB - Neoadjuvant chemotherapy is obligatory for small-cell lung cancer. When combined with radio-therapy, it reduces the incidence of recurrence and increases the survival rate to a level similar to that seen in non-small-cell lung cancer. Preliminary results suggest that complete remissions (3-year-survival rate 56%) can be achieved through the use of chemotherapy, possibly including high-dose chemotherapy for advanced stage III A cancer. The use of pre-operative chemotherapy in advanced stage III non-small-cell lung cancer is firmly established. In one study the 3-year-survival rate reached 25% in the chemotherapy group as compared to 15% in the group treated by surgery only. Early results of pre-operative chemo- and radiotherapy for stages III A and III B are encouraging. In studies comparing neoadjuvant chemotherapy alone to a combination of neoadjuvant chemo-radiotherapy a higher resection rate (32 to 76%) as well as a longer disease-free survival time could be shown for the combination therapy. To date, a number of innovative neoadjuvant chemoradiotherapy protocols using new substances as well as various modifications of radiotherapy are being studied. It is to be expected that new standard therapies for non-small-cell lung cancer will develop from these studies. PMID- 10829311 TI - [Preoperative radiochemotherapy of esophageal carcinoma. Light at the end of the tunnel?]. AB - Studies on neoadjuvant therapy of esophageal cancer showed that not either preoperative chemotherapy or radiotherapy lead to a significant improvement of prognosis. However, two prospective studies showed a significant prognostic improvement after neoadjuvant combined radio/chemotherapy. Most treatment protocols include a radiation with 30-45 Gy and a simultaneous therapy with Cis Platin/5-Fluorouracil. As an increase of perioperative morbidity and mortality has to be expected through this treatment, a careful selection of patients is necessary. Several studies have shown that mostly patients with good response benefit from this neoadjuvant therapy. The clinical response evaluation is difficult and response is best proved by classification of the histomorphologic regression of the tumor. For future research, predictive response analyses based on molecular biologic and immuno-histochemic techniques are of great importance and first differentiations by biomarkers have been detected. PMID- 10829312 TI - [Primary lymphoma of the stomach. An interdisciplinary challenge]. AB - Non-Hodgkin-lymphomas of the stomach are mostly of mucosal origin. The lymphoma derives from the acquired lymphatic tissue due to an infection with helicobacter pylori. Beside the grade of malignancy (low/high grade) the extent of the disease is the basis for therapy. It can be limited to the stomach and the lymph node compartments I and II or disseminated to both sides of the diaphragm. Therefore, pre-therapeutic diagnostic procedures have to cover all these localisations including the bone marrow. In case of a tumor limited to the stomach and of low grade malignancy an eradication therapy is the treatment of choice. More advanced tumors, but only involving stomach and lymph node compartments I and II, should be treated by subtotal or total gastrectomy plus lymphadenectomy according to the gastric cancer standards (D2). After R0-resections no adjuvant radio- or chemotherapy is necessary. R1/2 resections require radio- and/or chemotherapy which are the treatment of choice in a priori advanced stages and/or high grade lymphomas. The total 5 year survival rate is about 60-70%. Gastric lymphomas are an outstanding example of interdisciplinary disease management. PMID- 10829313 TI - [Preoperative downstaging in advanced stomach carcinoma. Wishful thinking or reality]. AB - By the time it is diagnosed, gastric carcinoma is usually already advanced and, as a result, has a poor prognosis. Surgery, with complete (R0) resection of the tumor, is the only chance of cure for this disease. However, in locally advanced gastric carcinoma this is only possible in approximately half of all cases. In order to help improve the prognosis of patients with advanced stage carcinomas, the concept of multimodal therapy is presently being evaluated. The results of studies of postoperative adjuvant therapy have been contradictory, with the result that no indication for such treatment outside of study protocols presently exists. Recently, preoperative application of chemotherapy, the so-called "neoadjuvant" therapy concept, has become increasingly important, since it has been demonstrated that, in individual cases, tumors thought to be primarily unresectable have been able to be completely resected after chemotherapy. Based on the available studies, one can assume that, in a subgroup of patients with not yet identified favorable tumor biologic characteristics, a true down staging of the tumor occurs. To what extent a preoperative "over-staging" may be a factor can only be estimated statistically, since the presently available methods for clinical estimation of tumor stage are never as accurate as the final histopathologic evaluation. Since the recently started, randomized multicenter study under the auspices of the EORTC compares surgery alone with a combination of surgery and preoperative chemotherapy in locally advanced gastric carcinoma, information will soon be available which will help clarify the effectiveness of this therapy concept. PMID- 10829314 TI - [Utilization of multimodal therapy concepts in stomach carcinoma in Germany]. AB - INTRODUCTION: In view of disappointing results after surgery alone multimodal therapeutic regimes are used to improve long-term prognosis in locally advanced gastric carcinomas. In presence of many reports about encouraging results ("down staging", improved R0-resection rates) but simultaneously missing evidence of efficiency of neoadjuvant therapies in respect to long-term survival (large randomized multicenter trials do not exist until today) and the herewith related uncertainties, we started an inquiry among many surgical units with the intention to evaluate the clinical practice of multimodal treatment for gastric cancer patients in Germany today. METHODS: In a questionnaire (3/99) we asked among 97 surgical units (41 university hospitals, 56 big community hospitals) in Germany for the management of gastric cancer patients with special interest to practice and state of adjuvant and neoadjuvant therapeutic strategies. Further we analyzed all resected gastric cancer patients (1986-1995) without neoadjuvant treatment in advanced stage of disease (pT3/4NxMx; stage III/IV (UICC'92) in respect to R0 resection rate and long-term prognosis (Kaplan-Meier). RESULTS: Overall feedback amounted to 78% (76/97) and was higher in university hospitals (90%) than in big community hospitals (70%). Today, neoadjuvant therapies are of more interest than adjuvant therapeutic regimes. But also neoadjuvant therapy is only used in 32% as a rule (in 16% with, in 16% without study conditions). 25% of all surgical units do not employ any neoadjuvant therapy in locally advanced gastric cancer until today. In all other surgical units neoadjuvant treatment is performed more individually and sporadically (43%) only in some patients. Neoadjuvant therapies are practiced by haematooncologists in 50%, gastroenterologists in 32% and surgeons in 27%. The predominant neoadjuvant therapeutic strategy is chemotherapy alone (84%). Many surgical units in Germany are interested to participate in a multicenter trial with more interest in neoadjuvant than adjuvant therapy. 185 of 309 resected gastric cancer patients (60%) were classified as stage IIIa, stage IIIb or stage IV patients. R0-resection rate of these advanced gastric cancer patients amounted to 37%; only 24% of them survived 5 years or more. CONCLUSIONS: Considering the missing evidence that multimodal therapies are able to prolong long-term survival in advanced gastric cancer patients, its use without study conditions is questionable. Conclusions, taken from data of clinical trials regarding carcinomas of the esophagus and esophagealgastric junction, are inconsistent in respect to long-term prognosis and results are not transferable to gastric carcinomas. A prospective randomized multicenter trial in advanced gastric cancer patients is of great importance. Following our data, in Germany a high readiness to participate in the forthcoming EORTC-study is present. PMID- 10829315 TI - [Neoadjuvant and adjuvant therapy of ductal pancreatic carcinoma]. AB - The optimal oncological management of ductal pancreatic cancer remains undefined. More than 60% of these patients have disseminated disease at the time of presentation. Here radical surgery alone cannot guarantee a cure. Even in the best case of a R0-resection with extended lymph node dissection the reported 5 year survival rates of 20-30% are dissatisfying. This would suggest that neoadjuvant or adjuvant therapies may play an even greater role in improving the medium and long-term survival rates than in other tumor entities. Reports in the literature to date are from small randomised trials which do not elucidate the benefit of therapy. However, it does appear that neoadjuvant radiochemotherapy in combination with R0-resection will best improve patient outcome and mean survival rates. Therefore there is a need for large prospective randomized studies regarding (neo-)adjuvant therapy. Inclusion criteria must be precisely defined and the following factors recorded: standardized preoperative staging procedures extent of tumor disease (histology, stage, vascular infiltration, lymph node involvement, etc.) detailed surgical approach in respect to the extent of pancreas resection and lymph node dissection. Pancreas resections tend to have a higher postoperative complication rate when compared with other tumours and substantial postoperative weight loss often is observed. This may result in a delay or even impossibility of starting adjuvant therapy right in time in a relevant part of patients (up to 1/3 according to literature data) which is a major disadvantage of all adjuvant therapy concepts. PMID- 10829316 TI - [Preoperative radiochemotherapy of rectal carcinoma. Current status]. AB - BACKGROUND: In locally advanced rectal cancer RO-resectability is limited and outcome is combined with a relatively high rate of local recurrences. This paper focuses on experiences with preoperative (neo-)adjuvant therapy and the potential improvement of treatment results. PATIENTS AND METHODS: Different therapy regimen were demonstrated under consideration of therapeutic induced side effects, treatment response (downstaging) in relation to resectability, recurrence free and overall survival. RESULTS: The rate of treatment induced toxicity of preoperative radio-(chemo-)therapy in locally advanced rectal cancer is acceptable with regard to the obtained treatment results. Through the pretreatment it was possible to gain a downstaging by nearly 60%. The frequency of local recurrence is significantly reduced by preoperative radiotherapy, and combined radiochemotherapy possibly increases the disease free survival. In what extent the results of treatment could be improved through hyperthermia and/or consecutive postoperative chemotherapy, is proved at present. CONCLUSIONS: Preoperative radio-(chemo-)therapy should be increasingly introduced into the standard treatment regimen of locally advanced rectal cancer and has to be optimised within the scope of further studies. PMID- 10829317 TI - [Are there still surgical indications in therapy of anal carcinoma?]. AB - The development of combined radio-chemotherapy according to Nigro has had a big influence on the therapy concept of the anal canal carcinoma. Due to the successful oncological treatment combined with a possible continence preservation, a rectum extirpation is rarely indicated. Surgical indications in the therapy concept of anal canal carcinoma are large, sphincter destroying tumors, treatment complications, therapy resistant progression and the therapy of lymph node- and organ metastases. In the therapy concept of the cancer of the anal margin, the local excision as a primary therapy is of central interest in the current literature. The introduction of plastic surgery in the treatment of even vast tumors leads to adequate results of the oncological and functional treatment. The abdomino-perineal rectum extirpation is exceptionally indicated for large, sphincter destroying tumors. PMID- 10829318 TI - [Methods for postoperative evaluation of complete excision of the mesorectum]. AB - This study aimed at a more objective evaluation of the specimen after total mesorectal excision [14]. For this reason, a method yielding a simple stained preparation of the totally excised mesorectum was developed. By postoperative injection of 10 ml of an ink solution into the A. rectalis superior of 15 specimens, the arterial mesorectal vascular tree was filled. All specimens had been collected by means of total mesorectal excision. In two specimens, in wich the mesorectal sheath fascia had been injured due to the surgical manipulation, we observed the leakage of ink from the mesorectum even during the injection. In three further specimens, some ink leakage in the form of dots occurred from small opened arterioles after the injection was performed. No ink leakage was observed in the remaining specimens. Prior to the ink injection, thirteen specimens were macroscopically tested and found intact. Three of the fifteen specimens exhibited minor lesions of the mesorectum that would not have been detected macroscopically without ink tagging. The comparison of the findings provided by the surgeon with the histopathological evaluation showed that those specimens in which no ink leakage occurred had an unimpaired mesorectal sheath fascia. These specimens coorespond to the complete excision of the mesorectum and the removal of the tumor in a cancer-sealed package as long as the circumferential rim of the specimen has not been infiltrated by the tumor. PMID- 10829319 TI - [Volumetry of pleural effusion in multi-morbidity, postoperative patients of a surgical intensive care unit. Comparison of ultrasound diagnosis and thoracic bedside image]. AB - Aim of this study was to evaluate the importance of chest ultrasound and chest x ray for the indication of thoracic drainage of pleural effusions in patients of an operative intensive care unit. Between December 1996 and June 1997 21 patients were included in a prospective trial in the operative intensive care unit. 26 thoracic drainages were used to drain pleural effusions. In all patients chest radiography in supine position and chest ultrasound were performed to assess the need of pleural drainage. Pleural fluid measured radiologically was categorized into 3 groups: pleural fluid less than 500 ml, 500 to 1,000 ml or more than 1,000 ml. The amount of the pleural effusion was sonographically determined by a standardized formula. After complete drainage of the pleural space the real volume of the fluid was measured and compared with the estimated value. The real amount of the fluid was correctly determined by chest radiographs in 16 cases (62%) and by chest ultrasound in 18 patients (69%). Pleural effusions less than 600 ml sonographically correlated much better with the real amount of the fluid than pleural effusions above 600 ml. In 8 cases (31%) ultrasound provided an additional information for correct indication of drainage. Considering both x-ray of the chest in supine position and chest ultrasound the correct indication to drain the pleural effusion was achieved in 25 cases (96%). In this prospective trial we compared chest ultrasound and chest radiography and demonstrated that ultrasound is more suitable to determine the amount of pleural effusions than radiography. In case of clinical and radiological suspicion on pleural effusion demanding for drainage a chest ultrasound should be performed to avoid underestimation of pleural fluid. PMID- 10829320 TI - [Effect of preoperative preoperative visualization therapy on postoperative outcome after inguinal hernia surgery and thyroid resection]. AB - This randomized controlled trial aims to evaluate the influence of preoperative relaxation techniques on postoperative outcomes. From January 1997 to June 1998 208 patients were operated on for primary inguinal hernia or goiter. The patients were randomized into two groups: Group A (n = 103) underwent the surgical treatment with a preoperative visualization therapy. Group B (n = 105) underwent the surgical treatment without a preoperative therapy. Patients with age under 18 years, ASA-status IV-V, recurrent inguinal hernia or recurrent goiter and malignant neoplasms were excluded from the study. There were no differences in age, sex, duration of the operation, training of the surgeon, and preoperative blood parameters between the two groups. RESULTS: During the postoperative follow up we observed more hematomas (group A with visualization therapy: 30.3%, group B without visualization therapy: 44.4%) as well as more pain (group A: 4.2, group B: 5.2) and analgesic consumption (group A: 59.7 mg Tramadol HCL, group B: 72.5 mg Tramadol HCL) in group B (p < 0.05). There were no significant differences in infections, nausea, hypocalcemia, tetania, recurrent nerve palsy, fever. CONCLUSIONS: Preoperative visualization therapy reduces significantly the number of postoperative hematomas after inguinal hernia repair. Furthermore a decrease of analgesic requirements after surgical treatment was observed. PMID- 10829321 TI - [Two scientific controversies shaping the development of surgery in the 20th century]. AB - By way of example, two scientific controversies that played a decisive role in shaping and determining the development--in the areas of chest and trauma surgery -of 20th century surgery, were discussed. In the former area, the spectacular method involving the use of a negative-pressure chamber, developed by Sauerbruch in 1904 was described--an innovation representing an enormous step forward in the field of surgery on the chest. However, the method failed to find widespread favour, and the opposite concept involving the use of positive-pressure was developed, and intratracheal ventilation--already under discussion at the beginning of the 20th century--finally won the day, and still remains in use. Medullary nailing of bone fractures as introduced by Kuntscher initially prompted controversial discussion and at first appeared to have been rejected--only to find widespread acceptance nevertheless. It continues to be a justified method of achieving a stable osteosynthesis which, thanks to methodological refinements and improvements over the years, is an important option in the list of indications for surgical treatment of bone fractures. All in all, the following remark would appear applicable: An achievement alone does not suffice--someone is needed to recognize and endorse it. PMID- 10829322 TI - [The upside-down stomach. Laparoscopic treatment is possible]. AB - An upside-down-stomach, usually occurring in eldery patients describes an extreme case of paraesophageal hernia. Due to the possibility of life threatening complications there is an urgent need for surgical intervention. In 1998 three patients suffering from complete upside-down-stomach were treated by laparoscopic surgery at our hospital. Two of them received a hiatoplastic with fundo- and corpophrenicopexy. In one case we did a 360 degrees-floppy Nissen's fundoplication. Perioperatively, one patient developed a left sided pneumothorax Long-term follow-up 6 months postoperatively in our out-patient department revealed unconspicuous clinical and gastroscopical findings and high patient satisfaction with postoperative outcome. Our case reports show the possibility of adequate operating on upside-down-stomach by means of laparoscopic surgery. Minimal invasive methods provide a good overlock on the operating field and are poor in complications when done by an experienced surgeon, so that patients may profit from comfort of laparoscopic surgery. PMID- 10829323 TI - [Symposium "Quality management in surgical oncology". Evaluation by speakers]. PMID- 10829324 TI - [Surgical proctology (2)]. PMID- 10829325 TI - Cannabis use and public health: assessing the burden. PMID- 10829326 TI - A longitudinal study of cannabis use and mental health from adolescence to early adulthood. AB - AIMS: To examine the longitudinal association between cannabis use and mental health. DESIGN: Information concerning cannabis use and mental health from 15 to 21 years was available for a large sample of individuals as part of a longitudinal study from childhood to adulthood. PARTICIPANTS: Participants were enrolled in the Dunedin Multidisciplinary Health and Development Study, a research programme on the health, development and behaviour of a large group of New Zealanders born between 1 April 1972 and 31 March 1973. MEASUREMENTS: Cannabis use and identification of mental disorder was based upon self-report as part of a general assessment of mental health using a standard diagnostic interview. Daily smoking and alcohol use at age 15 were assessed by self-report. Indices of family socio-economic status, family climate and parent-child interaction were formed using information gathered from parent report and behavioural observations over early childhood. Childhood behaviour problems were assessed by parent and teacher report. Attachment to parents was assessed in adolescence. FINDINGS: Cross-sectional associations between cannabis use and mental disorder were significant at all three ages. Both outcome variables shared similar pathways of low socio-economic status and history of behaviour problems in childhood, and low parental attachment in adolescence. Mental disorder at age 15 led to a small but significantly elevated risk of cannabis use at age 18; by contrast, cannabis use at age 18 elevated the risk of mental disorder at age 21. The latter association reflected the extent to which cannabis dependence and other externalizing disorders at age 21 were predicted by earlier level of involvement with cannabis. CONCLUSIONS: The findings suggest that the primary causal direction leads from mental disorder to cannabis use among adolescents and the reverse in early adulthood. Both alcohol use and cigarette smoking had independent associations with later mental health disorder. PMID- 10829327 TI - Does cannabis use encourage other forms of illicit drug use? AB - AIMS: To examine the relationship between cannabis use in adolescence and the onset of other illicit drug use. METHOD: Data were gathered over the course of a 21-year longitudinal study of a birth cohort of 1265 New Zealand children. Measures analysed included: (a) frequency of cannabis use and other illicit drugs from 15-21; (b) family, social, educational and behavioural backgrounds of cohort members prior to 15; and (c) adolescent life-style variables. FINDINGS: (i) By 21, nearly 70% of cohort members and used cannabis and 26% had used other illicit drugs. (ii) In all but three cases, the use of cannabis had preceded the use of illicit drugs. (iii) Those using cannabis on more than 50 occasions a year had hazards of other illicit drug use that were 140 times higher than non-users. (iv) After adjustment for covariate factors, including childhood factors, family factors and adolescent life-style factors, cannabis use remained strongly related to the onset of other forms of illicit drug use. Those using cannabis on more than 50 occasions per year had hazards of other illicit drug use that were 59.2 times higher than non-users. CONCLUSIONS: Findings support the view that cannabis may act as a gateway drug that encourages other forms of illicit drug use. None the less, the possibility remains that the association is non-causal and reflects factors that were not adequately controlled in the analysis. PMID- 10829328 TI - Youth escaping limits on drinking: binging in Mexico. AB - AIMS: In Tijuana, Mexico, a loosely enforced age-18 law and inexpensive drinks have given rise to a nightclub district frequented by thousands of young Southern Californians each weekend night. Surveys were designed to characterize the extent of the cross-border binge-drinking traffic and to support and evaluate the community's response. DESIGN: Over 1 year, two anonymous and voluntary breath test surveys were done. Drivers and pedestrians were pulled randomly from the stream of northbound border crossers and recruited to participate. SETTING: Surveys occurred between 12 a.m. and 4 a.m. on randomly selected Wednesdays, Fridays, and Saturdays. Participants were recruited within the Port of Entry building. PARTICIPANTS: Of the 5849 border crossers recruited, 87.4% participated in the survey. MEASUREMENTS: Information was obtained through a standardized verbal interview. All participants were asked to take an alcohol breath test. FINDINGS: On weekend nights, more than 6500 people cross back into the United States between 12 a.m. and 4 a.m. after drinking or visiting a bar or restaurant. Pedestrians represent the highest concentration of drinkers, with more than 30% having BACs of 0.08 or greater. Most of these pedestrians return to parked vehicles on the US side and drive or ride home. CONCLUSIONS: The flow of young binge drinkers at the Tijuana border is substantial and translates into a significant public health problem for the region as crossers use their vehicles to drive home. There are many such binge-drinking locales. However, the border is unique in that it is amenable to scientific estimations of the problem with relatively high precision. PMID- 10829329 TI - Natural disasters and alcohol consumption in a cultural context: the Great Hanshin Earthquake in Japan. AB - AIMS: To assess changes in alcohol consumption attributable to the Great Hanshin Earthquake to test the assumption that alcohol consumption increases after natural disasters. METHODS: Quarterly alcohol sales figures were compared for three periods: before, immediately after and subsequent to the Great Hanshin Earthquake in three areas of the Hyogo prefecture: the severely affected area, the moderately affected area and the unaffected area. Possible confounding by population movement, damage to retail outlets and normal variation in sales, was assessed. FINDINGS: The quantity of alcoholic beverages consumed in the heavily damaged areas as well as throughout the prefecture decreased from the 1994 pre disaster level, both immediately after the Great Hanshin Earthquake (January March 1995) and 2 years after the disaster. This finding remained once possible confounding factors were taken into consideration. CONCLUSIONS: More attention should be placed on drinking in the cultural context where a disaster occurs. It may sometimes deter, rather than encourage, drinking among the affected population. PMID- 10829330 TI - The use of drinking places by gender, age and ethnic groups: an analysis of routine drinking activities. AB - AIMS: This paper extends a prior analysis of drinking patterns to consider the influence of non-economic variables on the selection of drinking locations. DESIGN: Using data from a general population telephone survey conducted as a part of the Community Trials Project, Tobit models are estimated to determine the influence of background demographic characteristics upon the selection of drinking locations net of other model control variables. PARTICIPANTS AND SETTING: 24,778 current drinkers from four California and two South Carolina communities. FINDINGS: Distinct patterns of premise utilization are found to be associated with age, gender and ethnic subgroups. Additionally these patterns of utilization are differentially linked to drinking and driving, suggesting that patterns of outlet utilization are differentially linked to acute drinking problems (e.g. drunken driving and alcohol-related car crashes). CONCLUSIONS: Observed differences in outlet utilization patterns between age, gender and ethnic subgroups imply that preventive interventions should take into account the manner in which these subpopulations make use of drinking venues. PMID- 10829331 TI - A latent class analysis of antisocial personality disorder symptom data from a multi-centre family study of alcoholism. AB - AIMS: To determine if there are subtypes of Antisocial Personality Disorder (ASPD), as manifested by distinctive symptom profiles or by associations with alcohol, other drug dependence or other psychiatric disorders. METHODS: Data on 38 symptoms of ASPD (including childhood conduct disorder) obtained from probands, their relatives and controls (2834 females and 3488 males) recruited for the Collaborative Study on the Genetics of Alcoholism (COGA) were analyzed using latent class analysis. Associations of the resulting latent classes with alcohol dependence (AD) and other psychiatric disorders were examined. FINDINGS: Among women, a 4-class solution was obtained in which conduct disorder and ASPD were found almost exclusively in the most severely affected class with two additional classes with mild and moderate behavior problems also identified. A strong linear trend (p < 0.001) for AD was observed, with each successive class manifesting a higher prevalence than the previous class. Milestones of drinking careers and dependence on other drugs also showed a strong association with class severity. Among men, a 5-class solution was obtained and, like women, the highest prevalence of ASPD (74.6%) was found in the most severely affected class. Somewhat unexpected was the observation that prevalence of AD was equivalent in the two most severe classes. The data for men indicated a class with a milder spectrum of childhood misbehaviors but with an adult ASPD profile--as well as other psychiatric co-morbidity--that was comparable to the most severe class. CONCLUSIONS: Overall, findings from both men and women did not support the existence of subtypes of ASPD, but rather indicated a disorder distributed on a severity spectrum. PMID- 10829332 TI - Fatal heroin overdoses resulting from non-injecting routes of administration, NSW, Australia, 1992-1996. AB - AIMS: To document cases of fatal heroin overdose in New South Wales by non injecting routes of administration, and to compare the characteristics and toxicology of these cases with injecting fatalities. DESIGN: Examination of coronial files. SETTING: New South Wales, Australia. PARTICIPANTS: All fatal heroin overdose cases in NSW between 1992 and 1996. FINDINGS: There were 10 cases of death resulting from non-injecting routes of heroin administration between 1992 and 1996, representing 1% of cases. In three cases the route of administration was by inhalation, in five cases by nasal administration and in two cases by swallowing. The mean age of cases was 29.6 years, and nine of the cases were male. The median blood morphine concentration of non-injectors was 0.31 mg/l (range 0.06-0.99 mg/l). Drugs other than morphine were also detected in seven cases. CONCLUSIONS: Heroin overdose deaths are not restricted to the injection of heroin. While injection may constitute a greater overdose risk factor, there is no safe, overdose-free way to use heroin. PMID- 10829333 TI - Cognitive, dissociative and psychotogenic effects of ketamine in recreational users on the night of drug use and 3 days later. AB - AIMS: Recreational use of the N-methyl-D-aspartate (NMDA) receptor antagonist, ketamine, is increasing. The present study aimed to examine both the acute and residual effects of this drug on cognitive function, dissociation and schizotypal symptomatology in recreational users. DESIGN AND PARTICIPANTS: A parallel group design was used to compare 20 volunteers who reported having taken ketamine with 19 volunteers who reported no consumption of ketamine on the relevant night (day 0). All 39 participants were tested on day 0 and again 3 days later. On each test occasion a battery of tests was administered which tapped a wide range of memory functions, attention, dissociation, schizotypal symptomatology and mood. FINDINGS: Groups were broadly matched for polydrug use apart from ketamine. Acute effects on day 0 replicated previous laboratory studies showing a broad spectrum of cognitive impairments following ketamine administration as well as marked dissociative effects and schizotypal symptomatology. Three days later, ketamine users had significantly higher scores than controls on both dissociation and schizotypal symptomatology. On some cognitive measures there were no group differences on day 3; however, on tests tapping semantic memory, the ketamine users showed persisting impairments compared with controls. CONCLUSIONS: Ketamine appears to induce acute and severe impairments of working, episodic and semantic memory as well as psychotogenic and dissociative effects. Three days after drug ingestion, recreational users display semantic memory impairment and dissociative and schizotypal symptomatology which could reflect chronic or residual effects of taking the drug or pre-existing differences in ketamine users. PMID- 10829334 TI - Methadone maintenance and cessation of injecting drug use: results from the Amsterdam Cohort Study. AB - AIMS: To assess relationships between characteristics of methadone maintenance treatment and long-term cessation of injecting (> or = 1 year). DESIGN AND PARTICIPANTS: The incidence of cessation of injecting and relapse from non injecting to injecting was estimated among 488 participants of the Amsterdam cohort study among drug users. We used a nested matched case-control design to identify methadone treatment characteristics significantly and independently related to cessation of injecting. To ensure detailed and valid assessment of methadone treatment, data of the Central Methadone Register were linked with cohort data. For 339 of 488 subjects of the initial study group methadone data were available. FINDINGS: The incidence of cessation of injecting increased from 2.2/100 person years in 1985-89 to 5.5/100 per year in 1995-97 (Ptrend = 0.005). Relapse to injecting was high: 17.2/100 person years (no trend). Methadone dosage and frequency of methadone programme attendance in themselves were not significantly related to cessation of injecting. However, an individual increase of 5 mg or more per year (OR 4.20, 95% CI 1.54-11.46) and receiving methadone mainly via the outpatient clinic for drug-abusing prostitutes and foreigners (OR 0.18, 95% CI 0.05-0.59) were independent predictors of cessation of injecting. After cessation of injecting, there were no HIV-seroconversions during the period of non-injecting (129 person years). After relapse to injecting there was one seroconverter; however, follow-up was small (23 person years). The HIV-incidence of those who continued injecting was 3.2/100 per year. CONCLUSIONS: Steadily increasing the methadone dosage in a harm reduction setting may be useful in supporting injecting drug users in the process of cessation of injecting and reducing the spread of HIV-infection. PMID- 10829335 TI - The effectiveness of the Minnesota Model approach in the treatment of adolescent drug abusers. AB - AIMS: The treatment outcome of drug-abusing adolescents treated with a 12-Step approach. DESIGN: The study compares drug use outcome data at 6 and 12 months post-treatment among three groups of adolescents: those who completed treatment, those who did not and those on a waiting list. Also, among treatment completers, residential and outpatient samples were compared on outcome. SETTING: The treatment site is located in the Minneapolis/St Paul area of Minnesota. PARTICIPANTS: Two hundred and forty-five drug clinic-referred adolescents (12-18 years old), all of whom met at least one DSM-III-R substance dependence disorder. One hundred and seventy-nine subjects received either complete or incomplete 12 Step, Minnesota Model treatment and 66 were waiting list subjects. MEASUREMENTS: In addition to demographics and clinical background variables, measures included treatment involvement, treatment setting and drug use frequency at intake and follow-up. FINDINGS: Absolute and relative outcome analyses indicated that completing treatment was associated with far superior outcome compared to those who did not complete treatment or receive any at all. The percentage of treatment completers who reported either abstinence or a minor lapse for the 12 months following treatment was 53%, compared to 15 and 28% for the incompleter and waiting list groups, respectively. CONCLUSIONS: Favorable treatment outcome for drug abuse was about two to three times more likely if treatment was completed. Also, there were no outcome differences between residential and outpatient groups. Alcohol was the most common drug used during the follow-up period, despite cannabis being the preferred drug at intake. PMID- 10829336 TI - Development and validation of the Attitudes Towards Smoking Scale (ATS-18). AB - AIM: To develop and test the validity of a scale measuring attitudes towards smoking in current and former cigarette smokers. DESIGN AND PARTICIPANTS: In a first mail survey, we collected qualitative data from 616 smokers. In a second mail survey, we collected quantitative data from 529 smokers and ex-smokers. We conducted a 16-month follow-up survey among 93 participants in the second survey. SETTING: Geneva, Switzerland, 1995-98. FINDINGS: The study resulted in a three dimensional, 18-item scale: the "Attitudes Towards Smoking Scale" (ATS-18). The scale was validated with reference to criteria of content-, construct- and predictive validity. The three subscales measure perceptions of adverse effects of smoking (10 items), psychoactive benefits (four items) and pleasure of smoking (four items). Internal consistency coefficients (0.85, 0.88 and 0.81) and test retest correlations were high (0.90, 0.75, 0.89, respectively). Differences in attitude scores between smokers in the pre-contemplation and preparation stages of change were -0.83, 0.71 and 1.23 standard deviation units, respectively. A differential score (advantages minus disadvantages of smoking) predicted smoking cessation in baseline smokers and relapse in baseline ex-smokers. CONCLUSION: ATS 18 is a valid and reliable instrument which can be used in both research and clinical settings. PMID- 10829337 TI - A new force--but for what and whom? The appropriation of networks and the importance of decision rules. PMID- 10829338 TI - A microtitre plate assay for the detection of antibiotics in porcine urine. AB - Techniques for screening porcine samples for antimicrobial residues in the EU usually involve analysis of samples taken post slaughter, and are either time consuming or expensive. Some of the positive test results at this screening stage could be avoided by allowing the animal sufficient withdrawal time following drug treatment. A method is described that can detect the presence of five major antibiotics in porcine urine at concentrations below 1 microgram ml-1 for each of the compounds. The test uses Bacillus subtilis, which is already widely employed in antimicrobial inhibition assays, and when combined with a colorimetric substrate, p-nitrophenyl-beta-D-glucopyranoside, can detect inhibitory substances within an assay time of four and a half hours. The method, which uses microtitre plate technology, could be developed into a convenient test kit for use at farm level to determine whether animals were still excreting antimicrobials in their urine prior to their submission for slaughter. PMID- 10829339 TI - Separation and enrichment of palladium and gold in biological and environmental samples, adapted to the determination by total reflection X-ray fluorescence. AB - The reductive co-precipitation of trace and ultra-trace elements together with mercury followed by complete evaporation of the mercury makes it possible to determine palladium and gold by total reflection X-ray fluorescence. Both elements can be detected without interferences at optimal sensitivity in the pg range. Thus, detection limits of, e.g., 2.5 ng L-1 for palladium and 2.0 ng L-1 for gold, in urine, were obtained. The precision was determined to 0.04 at a palladium concentration of about 200 ng L-1 urine and to 0.19 at a gold concentration of only 18 ng L-1. The recovery for a urine sample spiked with known amounts of palladium and gold amounted to > 95%. Results of the combined procedure are given for the determination of palladium and gold in the urine of non-exposed and occupationally exposed persons and in some other environmentally relevant samples. PMID- 10829340 TI - Determination and quantification of sulfadiazine and trimethoprim in swine tissues using liquid chromatography with ultraviolet and mass spectrometric detection. AB - High-performance liquid chromatographic procedures with ultraviolet detection were developed for the quantitative determination of sulfadiazine (SDA) and trimethoprim (TMP) in swine tissues (kidney, liver, muscle, fat and fat + skin). In addition, high-performance liquid chromatography with atmospheric pressure chemical ionization mass spectrometry was used for the confirmation of the identity of the analytes of interest. Chromatographic separation was achieved on a Spherisorb ODS-2 column (250 x 4.6 mm id, dp 5 microns). The mobile phase for SDA analysis consisted of 1% acetic acid in water-acetonitrile (85 + 15, v/v). For TMP analysis a 80 + 15 + 5 (v/v/v) mixture of 0.25% triethylammonium acetate in water, acetonitrile and methanol was used as the eluent. Sulfamerazine and ormethoprim were used as the internal standards for SDA and TMP analysis, respectively. For the isolation of the compounds of interest from biological samples, a liquid-liquid extraction with acetone and ethyl acetate, followed by a clean-up using a solid-phase extraction column (aminopropyl and benzenesulfonic acid for SDA, benzenesulfonic acid for TMP) was performed. Calibration graphs were prepared for all tissues and linearity was achieved over the concentration ranges tested (50-1000 ng g-1 for SDA, r > or = 0.9979; 25-500 ng g-1 for TMP, r > or = 0.9994). The method was validated at the maximum residue level (MRL, 100 ng g-1 for SDA and 50 ng g-1 for TMP), at half the MRL and at double the MRL for both SDA and TMP. The accuracy and precision (expressed as the within-day repeatability) were found to be within the required ranges for each specific concentration. The quantification limits were 50 ng g-1 for SDA and 25 ng g-1 for TMP. The limits of detection were below one half the MRLs. Both methods were selective for the determination of SDA and TMP. Biological samples (kidney, liver, muscle, fat and fat + skin) from pigs that received a commercial SDA-TMP preparation with the feed for five consecutive days (dose rate: 25 mg SDA and 5 mg TMP kg body weight-1 day-1) were analyzed using the described methods. The quantitative results were used to calculate a withdrawal time (12 days) to reach residue levels below the respective MRLs. This calculation was performed according to the recommendations of the European Agency for the Evaluation of Medicinal Products (EMEA/CVMP/036/95). PMID- 10829341 TI - HPLC determination of catechins and caffeine in tea. Differentiation of green, black and instant teas. AB - A simple and fast high performance liquid chromatographic method for five catechins and caffeine using an ODS column and a water-acetonitrile-formic acid mobile phase system was developed. The catechins (epicatechin, catechin, epigallocatechin, epigallocatechin gallate, epicatechin gallate) and caffeine were separated by an acetonitrile gradient within 20 min. The detection limit of the method was approximately 10 ng for all the compounds (by injecting 10 microL). Several green, black and instant teas were analysed using this method. By using the studied compounds as chemical descriptors, linear discriminant analysis was performed and complete differentiation of the green, black and instant teas was achieved. PMID- 10829342 TI - Simultaneous determination of fat-soluble vitamins and provitamins in dairy products by liquid chromatography with a narrow-bore column. AB - A reversed-phase high-performance liquid chromatographic method is described for the simultaneous determination of vitamins A, D2, D3, E and K1, retinyl acetate, retinyl palmitate, tocopherol acetate, ergosterol and 7-dehydrocholesterol in milk and butter. Narrow-bore columns are recommended because this alternative provides a good separation and efficiency, plus greater economy and sensitivity. Detection limits for individual vitamins range from 0.14 to 6.9 ng. All vitamins are separated in less than 33 min. For the simultaneous determination of these vitamins and provitamins we use two sample pre-treatment methods, a liquid-liquid extraction with hexane or a solid-phase extraction with a C18 cartridge. Recovery studies show good results for all solutes (84-108% and 85-108% for milk and butter, respectively) and the intra-day coefficients of variations range from 1.6 to 4.5%. These methods permit the simple determination of fat-soluble vitamins using a small sample volume. PMID- 10829343 TI - Bioavailability studies of oral dosage forms containing levodopa and carbidopa using column-switching chromatography followed by electrochemical detection. AB - A reliable multi-dimensional column chromatographic method employing amperometric detection using a carbon fibre microelectrode procedure was used for monitoring the plasma profiles and to evaluate the pharmacokinetics and bioavailability of levodopa (L-dopa) and carbidopa (C-dopa), after ingestion of oral formulations containing these drugs. The peak currents obtained for the different analytes were directly proportional to the analyte over the concentration range 0.02-4 micrograms ml-1. Using this method, the minimum detectable concentration was estimated to be 5 and 8 ng ml-1 for L-dopa and C-dopa, respectively. Recovery studies ranged from 93.83 to 89.76%, with a relative standard deviation of less than 7%. The study was carried out in two separate weeks on five healthy non patient fasted male/female volunteers in the age range 20-37 years and weighing between 60 kg and 78 kg. The pharmacokinetic profile of two controlled-release products containing both L-dopa and C-dopa (Sinemet CR3 and CR4) was compared on the one hand and Sinemet conventional tablets on the other. The pharmacokinetic parameters, peak concentration (Cmax), the time taken to obtain this level (Tmax), elimination half-time T1/2, elimination rate constant (Kel), plasma level ratio, fluctuation index (FI) and the area under the time-concentration curve (AUC0-8), were investigated for each individual formulation. A comparison of the uptake of L-dopa from the conventional formulation showed that L-dopa entered the plasma and achieved peak levels higher than that of the controlled release formulations. However, it showed a much higher fluctuation index and the plasma concentrations were more stable with the controlled release formulations. The data also indicated a very low accumulation of both levodopa and carbidopa following repeated administration of the drugs, which was consistent with their relatively short half-lives (less than 2 h). In contrast, the half-life for the metabolite 3-orthomethyl dopa (3-OMD) is in the order of 13 h. As a result, there was an extensive accumulation of 3-OMD and its levels were significantly higher than those of levodopa or carbidopa upon repeated administration. Urine recoveries of the three analytes over one 8 h dosing interval showed that the majority of the excreted levodopa and carbidopa was recovered during the first 4 h, and there is proportionally greater excretion of the carbidopa dose than the levodopa dose. PMID- 10829344 TI - Determination of inorganic anions in biological fluids with direct sample injection by electrostatic ion chromatography using zwitterionic micelles in both stationary and mobile phases. AB - A new ion chromatographic (IC) system, which uses zwitterionic (e.g., Zwittergent 3-14) micelles as both stationary and mobile phases, highly useful for the analysis of inorganic anions in biological samples, was developed. The zwitterionic micellar stationary phase (which is obtained by immobilizing the zwitterionic surfactant on surfaces of the reversed-phase ODS) showed high ability to confine the elution bands of the large amount of SO4(2-) and Cl- to narrow zones. As a result, a base-line separation of NO2-, Br- and NO3- from SO4(2-) and Cl- is always achieved. The zwitterionic micellar mobile phase, (which is obtained by dissolving the zwitterionic surfactant with a suppressive electrolytic solution, e.g., aqueous NaHCO3 solution), on the other hand, showed high ability for rapid elution of proteins. The separation column is therefore always being cleaned up even after the protein-containing sample is directly injected. The zwitterionic micelles are also insensitive to conductivity detection, therefore either the suppressed or the non-suppressed conductivity detection method is applicable for detection of the analyte ions. Urine and serum were chosen as the model real samples and were analysed with direct sample injection; the results of successful determination of a number of inorganic anions (SO4(2-), Cl-, NO2-, Br- and NO3-) in both samples have demonstrated the usefulness of this new IC system. PMID- 10829345 TI - Determination of iodide by derivatization to 4-iodo-N,N-dimethylaniline and gas chromatography-mass spectrometry. AB - A real-time determination of iodide is proposed which involves the oxidation of iodide with 2-iodosobenzoate in the presence of N,N-dimethylaniline. The reaction is completed within 1 min to yield 4-iodo-N,N-dimethylaniline, which is extracted in cyclohexane and determined by GC-MS. It was also possible to determine iodine by derivatization in the absence of 2-iodosobenzoate, and iodate by its reduction with ascorbic acid to iodide and subsequent derivatization. A rectilinear calibration graph was obtained for 0.02-50 micrograms l-1 iodide with a correlation coefficient of 0.9998. The limit of detection was 8 ng l-1 iodide. The method was applied to the determination of iodate in iodized table salt and free iodide and total iodine in sea-water, and to spiked samples when the recovery was in the range 96.8-104.3% (RSD 1.9-3.6%). A sample clean-up by solid phase extraction with a LiChrolut EN cartridge is proposed. PMID- 10829346 TI - Determination of proteins in serum by fluorescence quenching of rose bengal using the stopped-flow mixing technique. AB - The stopped-flow mixing technique was used to develop a very fast, sensitive and accurate method for determining total proteins. The method is based on the lower fluorescence of Rose Bengal caused by binding of the dye to the proteins. The decrease in the fluorescence intensity, measured at 572 nm with excitation 555 nm, was linearly related to protein concentration from 1.3 to 24.5 micrograms ml 1. The detection limit was 0.3 microgram ml-1. The method was satisfactorily applied to the determination of total proteins in different serum samples. PMID- 10829347 TI - Aromatic aldehydes as fluorogenic indicators for human aldehyde dehydrogenases and oxidases: substrate and isozyme specificity. AB - Substrate properties of a number of potentially fluorogenic aromatic aldehydes of naphthalenes, phenanthrenes and anthracenes and of some coumarin aldehydes towards various forms of the human and rat aldehyde oxidase and dehydrogenase were examined using absorption and emission spectroscopy. It was demonstrated that recombinant human class 1 aldehyde dehydrogenase (ALDH-1) readily oxidizes naphthalene (except for those ortho-substituted), phenanthrene and coumarin aldehydes, whereas the class 3 enzyme (ALDH-3) from human saliva is active only towards 2-naphthaldehyde derivatives. The observed reaction rates in both cases are comparable to those of the best known substrates, and the Km values are typically in the sub-micromolar range. Aldehyde oxidases (AlOx), which are present in mammalian liver, reveal much broader substrate specificity, oxidizing nearly all the compounds examined, including those of the anthracene series, with maximum activity in the micromolar range of substrate concentration. In rat liver, nearly all AlOx activity was located in the cytosolic fraction. PMID- 10829349 TI - Inhaler devices for asthma. AB - Inhalation therapy is now the mainstay of treatment for asthma but for many patients such treatment is still not optimal. The delivered dose can vary widely between different delivery systems, and between patients, depending on how well they use a particular device. Many new inhaler devices are now available, and their competing promotional claims can confuse both prescribers and patients. In this article we review the devices marketed and discuss how to select the best one for a particular patient. PMID- 10829348 TI - Rapid analysis of ecstasy and related phenethylamines in seized tablets by Raman spectroscopy. AB - Raman spectroscopy with far-red excitation has been used to study seized, tableted samples of MDMA (N-methyl-3,4-methylenedioxyamphetamine) and related compounds (MDA, MDEA, MBDB, 2C-B and amphetamine sulfate), as well as pure standards of these drugs. We have found that by using far-red (785 nm) excitation the level of fluorescence background even in untreated seized samples is sufficiently low that there is little difficulty in obtaining good quality data with moderate 2 min data accumulation times. The spectra can be used to distinguish between even chemically-similar substances, such as the geometrical isomers MDEA and MBDB, and between different polymorphic/hydrated forms of the same drug. Moreover, these differences can be found even in directly recorded spectra of seized samples which have been bulked with other materials, giving a rapid and non-destructive method for drug identification. The spectra can be processed to give unambiguous identification of both drug and excipients (even when more than one compound has been used as the bulking agent) and the relative intensities of drug and excipient bands can be used for quantitative or at least semi-quantitative analysis. Finally, the simple nature of the measurements lends itself to automatic sample handling so that sample throughputs of 20 samples per hour can be achieved with no real difficulty. PMID- 10829350 TI - Rivastigmine for Alzheimer's disease. AB - Rivastigmine (Exelon-Novartis) is the second cholinesterase inhibitor marketed for symptomatic treatment of mild to moderately severe Alzheimer's dementia, and follows [symbol: see text] donepezil (Aricept-Eisai; Pfizer). Previously, we have been "unconvinced of the value of donepezil in routine clinical practice". Rivastigmine has been promoted with the slogan "Beyond cognition: improving functional ability". Does rivastigmine offer useful benefits in Alzheimer's disease? PMID- 10829351 TI - Tackling myocardial infarction. AB - Myocardial infarction is dangerous. Among people in the UK who develop an acute coronary event, in particular myocardial infarction, around 35-40% die within 24 hours of the condition's onset and 40-50% within a month. Here, we discuss management up to and following admission to hospital, concentrating on the use of aspirin, thrombolytic therapy, coronary angioplasty, angiotensin-converting enzyme (ACE) inhibitors and beta-blockers. PMID- 10829352 TI - Corticosteroids for croup. AB - Croup (laryngotracheobronchitis) is a common childhood respiratory condition, characterised by a harsh, barking cough, a hoarse voice, stridor and variable respiratory distress. Corticosteroids are often used to treat the condition. Four years ago, we concluded that nebulised budesonide improved croup symptoms of stridor and cough (but not necessarily dyspnoea) but that more trials were needed to clarify its role. In this article, we reconsider the place of corticosteroids in croup and assess whether different products offer particular advantages. PMID- 10829353 TI - Massive pulmonary embolism: do algorithms aid management? PMID- 10829355 TI - Rapid diagnosis of massive pulmonary embolism in a district general hospital. AB - Effective treatment of massive pulmonary embolism is more likely if diagnostic tests are rapidly available, including during out-of-hours. An agreed protocol was implemented in November 1997, which allowed initiation of thrombolysis by junior doctors within an hour of clinical suspicion of the diagnosis of massive pulmonary embolism in six patients in the subsequent year. A similar approach could be considered by other acute hospitals. PMID- 10829354 TI - Home management of fever in children: rational or ritual? AB - Fever is extremely common in childhood. Parents have been shown to have unrealistic fears, resulting in inappropriate management of fever in their children. This study was conducted to survey parents about their knowledge concerning home management of fever in children in their care. Parents of 560 febrile children were randomly recruited and interviewed in the waiting areas of the outpatient clinics or emergency room in four hospitals in Riyadh city using a standard questionnaire. Most of the interviewees were mothers, aged within 20-39 years. Although more than one-half of fathers and one-third of mothers in the study were well educated, most were misinformed about recognition and definition of fever. Most parents had poor knowledge regarding minimum temperatures for administering correct doses of antipyretic drugs or for sponging/bathing with water of the correct temperature. Most parents demonstrated a poor understanding of the appropriate frequency for checking the child's temperature and administering antipyretics. Only one-third of parents indicated a reasonable educational imprint by health-care providers. Considerable efforts will be required to educate parents about fever and its management. PMID- 10829356 TI - Management of septicaemic infants during long-term parenteral nutrition. AB - Young infants, particularly following gastrointestinal surgery, are at high risk of septicaemia during parenteral nutrition. Febrile illness in the absence of focal infection inevitably raises suspicion of central venous catheter sepsis and poses the following dilemma: remove the catheter (which may then prove uninfected) and lose venous access, or leave the catheter and risk clinical deterioration? We examined retrospectively the isolates from blood culture during febrile episodes in 13 children who received long-term (> 2 months) parenteral nutrition via a central venous catheter, and assessed the effectiveness of through-catheter antibiotic treatment during 76 episodes of blood culture positive sepsis. Coagulase-negative Staphylococci accounted for only 16% of positive isolates, with yeasts accounting for 5%, and Gram-negative organisms accounting for 46%, suggesting that infection was often associated with bacterial translocation from the gastrointestinal tract. Treatment with the central venous catheter left in situ was successful in resolving infection in 53 (70%) of septic episodes. These findings indicate that, in this specific group of patients, through-catheter antibiotic treatment is often effective in treating septicaemia. When long-term venous access is essential, this approach should be tried before recourse to central venous catheter removal. PMID- 10829357 TI - Pernicious anaemia and neurophysiological studies in Arabs. AB - While pernicious anaemia is a disease predominantly affecting people of north European descent, there is increasing awareness of its occurrence in other ethnic populations. We document herein details of six Arabs who had pernicious anaemia and significant neurophysiological abnormalities. All the patients made a complete haematological and symptomatic recovery following parenteral administration of hydroxycobalamin. The diagnosis of pernicious anaemia should be considered in Arab patients of all ages who have macrocytic anaemia, or symptoms of peripheral neuropathy. We caution against the empirical use of folic acid and vitamin B12 in the management of patients with chronic anaemia. PMID- 10829358 TI - The whys and wherefores of getting patients to give up smoking. AB - Smoking-related diseases are responsible for the majority of premature deaths in industrialized societies. It is 50 years since the link between smoking and lung cancer was first established, since when a gradually increasing number of smoking related illnesses have been identified. The treatment of patients suffering smoking-related illnesses consumes a large proportion of health care budgets. The ever-increasing costs of health care, in part due to the advent of new therapeutic techniques, could be offset by a concerted effort to reduce the number of people smoking. Various strategies, particularly if used in combination, have been shown to produce acceptable smoking cessation rates. At present there appears to be a reluctance on the part of governments to fund a concerted, co-ordinated programme to encourage and assist people to stop smoking. In the absence of such support it is important for all health professionals to provide sensible advice and support to help their patients to stop smoking. PMID- 10829359 TI - Current management of multiple sclerosis. AB - Multiple sclerosis is a complex disease which presents a unique challenge in clinical management. This decade has seen the advent of new therapies which are partially effective in modifying the disease course. However, current therapies have no impact on existing neurological deficits and so supportive measures, symptomatic treatment and comprehensive rehabilitation remain at the core of management. Historically clinical practice has often been empirical but management should now be evidence-based and guided by consensus opinion if improvements in care are to be made. PMID- 10829360 TI - The current management of cystic fibrosis. AB - Cystic fibrosis (CF) is one of the commonest lethal inherited conditions among Caucasians. It affects multiple organ systems and exhibits a range of clinical problems of varying severity. Life expectancy has improved in recent years as treatment regimes have become more intensive, but current treatments are expensive, often time consuming and may affect quality of life. This review summarises the treatments currently used in the management of patients with CF, and the evidence for these. PMID- 10829361 TI - Doxazosin in elderly patients with hypertension. AB - Hypertension constitutes a major cardiovascular risk factor of high prevalence in the elderly, and reducing elevated blood pressure has been shown to be of significant benefit in decreasing the incidence of cardiovascular and cerebrovascular disease in this patient population. Elderly patients are more likely to have comorbid disorders, such as dyslipidaemia, diabetes, renal disease, atherosclerosis and, for males, benign prostatic hyperplasia (BPH). Therefore, when choosing an antihypertensive agent for elderly patients, it is particularly important to ensure that treatment does not exacerbate comorbid conditions and does not interact deleteriously with any concurrent medication that the patient is taking. The alpha 1-adrenoceptor antagonist, doxazosin, has been shown to be an effective, well-tolerated antihypertensive therapy in elderly male patients and does not exacerbate--and in some cases improves--some other common disorders. Doxazosin has been shown to be effective in reducing the symptoms of BPH in elderly patients whose blood pressure is well controlled by concomitant antihypertensive medication. In addition, improvements in the symptoms of BPH as well as reductions in blood pressure have been observed in elderly men with mild-to-moderate hypertension. Doxazosin has been shown to have positive effects on lipid profiles and glycaemic control, which make it an attractive choice of therapy for elderly patients with hypertension and diabetes or dyslipidaemia. In addition, doxazosin is administered once daily, either in the morning or the evening, which may aid compliance, an important consideration in the elderly. PMID- 10829362 TI - Afelimomab. AB - Tumour necrosis factor-alpha (TNF-alpha) is well established as a key mediator in the inflammatory response seen in various disease processes including sepsis. TNF alpha is involved in virtually all features of septic shock and multiple organ failure. Anti-TNF-alpha strategies are thus appealing and have been effective at reducing inflammation and morbidity in certain conditions including rheumatoid arthritis and Crohn's disease. Afelimomab is the F(ab')2 fragment of a murine anti-TNF-alpha antibody, and has been evaluated in clinical trials in septic patients. The results suggest that the drug is well tolerated, and may be of benefit in certain groups of patients with sepsis. A large, randomised, clinical trial of afelimomab in patients with severe sepsis has recently been completed and the results are eagerly awaited. More work is necessary to identify a means of selecting which patients are most likely to benefit from this type of therapy in sepsis. PMID- 10829363 TI - Dapsone-induced methhaemoglobinaemia: a source of diagnostic confusion. AB - Arterial desaturation as measured using pulse oximetry may not reflect cardiorespiratory disease; other possible causes, including certain drugs, should be sought. Within the literature, examples exist of dapsone-induced methaemoglobinaemia causing diagnostic confusion, particularly where respiratory disease is a feature. Few cases have been reported that demonstrate the potential of relatively low levels of methaemoglobinaemia to upset pulse oximetry readings. We describe three examples of dapsone-induced methaemoglobinaemia emphasising the potential for low-grade methaemoglobinaemia to cause diagnostic confusion. Widespread use of the pulse oximeter indicates this problem may occur more regularly, hence there is a need for increased awareness. PMID- 10829364 TI - Solitary plasmacytoma of the parotid gland. AB - Extramedullary plasmacytoma involving the parotid gland is a rare occurrence. This report describes a case of solitary extramedullary plasmacytoma arising from the parotid gland of a 60-year-old woman. The importance of thorough follow up for these patients is stressed, as the possibility of multiple myeloma is higher in this group. PMID- 10829365 TI - Osteoid osteoma of the scapular neck: a case report. AB - Osteoid osteoma localised in the scapula are very rare. We report the case of an 11-year-old girl, who presented with an osteoid osteoma at the neck of the glenoid. This was excised en bloc after being located accurately by computed tomography. This case report demonstrates the difficulty in the approach to the excision of this unusually located lesion. PMID- 10829366 TI - Chylous ascites: a rare complication of radical gastrectomy. AB - Chylous ascites is the accumulation of lymphatic fluid within the peritoneal cavity, due to trauma or obstruction to the lymphatic system. Postoperative chylous ascites is a rare complication of abdominal surgery. This is frequently reported after retroperitoneal dissections, and results in high morbidity and mortality. The treatment options are varied and include total parenteral nutrition (TPN), elemental diet with medium chain triglycerides (MCT), repeated paracentesis and surgical ligation. We report a case of post-operative chylous ascites after D2 distal gastrectomy. Treatment by fasting, TPN followed by fat free diet resulted in complete resolution of ascites within 3 weeks. To our knowledge this is the first report of such a complication following radical gastrectomy. We review the literature and briefly discuss the management options. PMID- 10829367 TI - Digital cataract photography. AB - This study compares the cost effectiveness of two methods of grading cataract changes in the Australian population. The conventional film-based method has comparatively low set-up costs, but is more subjective in its assessment and more reliant on manpower, while the digital method has high set-up costs, but the results are more objective and available more quickly using fewer people. By a careful assessment of all the costs involved it is possible to estimate how many participants need to be recruited to a study in order to make the digital method cost effective. PMID- 10829368 TI - An investigation into digital imaging in assessing cosmetic outcome after breast surgery. AB - This study investigates post-operative photographic assessment in determining the cosmetic outcome of 74 breast cancer patients who underwent a breast lumpectomy and radiotherapy. Using 10 of these patients picked at random, comparison was made between a conventional photographic print, a print produced from a digital image, and a digital image viewed on a computer screen in terms of personal preference for clinical assessment. The cosmetic outcome scores obtained on the basis of these images were compared with cosmetic outcome scores obtained by direct observation both by the clinician and by the patient. In the analysis of image preference, conventional prints scored highest, but each of the image types was considered to be acceptable for assessing breast cosmesis. Statistical analysis of the cosmetic outcome scores proved that there was a significant correlation between the scores obtained from the images and the scores obtained by direct observation both by the clinician and by the patient. PMID- 10829369 TI - The Trust experience: eight years on. AB - In 1992 the author described her experience of working in a newly formed acute NHS Trust hospital. One of the key messages of the paper concerned the pace of radical changes experienced. Eight years on, the process of change has accelerated still further, and healthcare professionals working in the NHS face newer and tougher challenges with regard to funding, recognition and survival. This paper describes the author's personal view of developments so far. PMID- 10829370 TI - A new visual identity for the National Health Service. AB - The following article gives a brief overview of the new visual identity being adopted by the National Health Service in England. It looks at the thinking behind the identity, the identity's component parts and provides sources for obtaining further information on the identity's application. It is compiled from a presentation by Stephanie Hood from the corporate identity team of the NHS Executive communications unit given on 22nd October 1999 at the National Designers in Health Network seminar, Time-out '99, Sheffield. Supporting information was obtained from the NHS Communications website http:?nww.doh.nhsweb.uk/commsnet. PMID- 10829371 TI - Colour perception in telemedicine. AB - A doctor's experience of examining patients is for the most part in artificial light, which is known to affect colour perception. However, the phenomenon of colour constancy (the perceived stability of the colour of objects, despite changes in the illuminating light) allows the eye and brain to compensate, to some extent. Colours like red, yellow, green and blue, which are processed by basic colour-opponent mechanisms, show better colour constancy than intermediate colours. These are the very colours that are involved in skin tones. A skin rash can therefore be viewed sometimes acceptably with lower-quality telemedical systems. Colour is undoubtedly important in telemedicine, but while a great deal is known about the fundamentals of colour perception from laboratory experiments, little is known about the effects of colour on the outcome of the telemedical process. The field of colour perception therefore represents a fertile area for telemedical research. PMID- 10829372 TI - Domiciliary midwifery support in high-risk pregnancy incorporating telephonic fetal heart rate monitoring: a health technology randomized assessment. AB - We conducted a health technology assessment of the care of women with high-risk pregnancies in the South Wales valleys. Women in the control arm were intended to receive conventional care with standard midwifery visits. Women in the intervention arm received additional or longer visits and domiciliary fetal heart rate telemonitoring. Eighty-one mothers were randomized. There were significant differences in midwifery intervention resources between domiciliary and control groups, with the former receiving a mean of 3.7 visits lasting 33.5 min, compared with 1.4 visits lasting 12.8 min for the latter. There were slightly more spontaneous labours and fewer Caesarean sections in the domiciliary group. Maternal satisfaction and anxiety were high in both groups. Domiciliary care increased the service costs by 21.02 Pounds per woman in terms of extra midwife travel and visiting time, and by a further 18.38 Pounds per woman in home monitoring equipment costs. This, however, was more than offset by health service savings from fewer clinic visits (35.60 Pounds) and fewer clinic ultrasound scans (9.01 Pounds). Adding the reductions in lost productivity to women and their partners (34.51 Pounds) suggests that domiciliary care was cheaper than conventional care, even if it did not greatly reduce inpatient days (a reduction nonetheless saving 184.24 Pounds). While clinical processes were similar in both groups, there were useful practical advantages and savings for patients and the health service from the domiciliary intervention. PMID- 10829373 TI - Factors that may influence the implementation of nurse-centred telemedicine services. AB - A qualitative study of four UK telemedicine sites was carried out to explore reactions to nurse-centred telemedicine services and their possible implications. Using semistructured schedules, 36 people directly involved in these services were interviewed--25 nurses, four general practitioners, two advising medical consultants, three service managers and two researchers. Factors identified which may influence successful implementation included: early involvement of all groups affected, prior consideration of the practical implications, thorough initial equipment testing, good technical support, imaginative training and clarity of the purpose of the service. The study showed that all those involved must be prepared for the rapid learning that is implicit in the implementation of change. The challenge is to maximize the potential for the primary operator to use new techniques. In respect of nurse-centred services, the nurse must not be viewed as a technician, nor should telemedicine be seen as a substitute for an available doctor. PMID- 10829374 TI - User satisfaction with realtime teleneurology. AB - User satisfaction (i.e. that of patients, medical staff at a remote hospital and medical staff at a neurological centre) with realtime teleneurology consultations was studied prospectively. Twenty-five patients with neurological problems admitted to a hospital without permanent neurological cover were assessed from a neurological centre by specialist neurologists using realtime video-links transmitting at 384 kbit/s. All users reported high levels of satisfaction with the technical aspects of the consultations. Patients, almost universally, reported confidence in teleneurology as a means of dealing with their presenting complaints. Similarly, medical staff at either site felt confident in managing patients using teleneurology and almost always felt that a telephone consultation would not have achieved as good an outcome. No major organizational problems were identified. These findings suggest overall user satisfaction with realtime teleneurology for managing patients with neurological problems admitted to hospitals that do not have resident neurologists. PMID- 10829375 TI - Telemedicine and cardiopulmonary resuscitation: the value of video-link and telephone instruction to a mock bystander. AB - We evaluated cardiopulmonary resuscitation (CPR) performed by persons with no previous experience on a resuscitation dummy. Subjects were randomized into four groups, one of which had no instruction. The other three groups were instructed for 3 min in mock CPR by a supervisor using a telephone, a video-link, or directly in person. They were compared with a group which had had previous CPR training. The main outcome measures were the number of correct ventilations, chest compressions and compressions with correct hand position. Video-link instruction was associated with significantly higher median scores for all three outcome measurements (P < 0.05), whereas telephone instruction and previous CPR training were associated with higher scores on only one, namely ventilations (P < 0.05). Video-link instruction was comparable with direct observer instruction. There was no significant difference between previously trained subjects and the intervention groups. Video-link instruction can produce significant improvements in the quality of CPR in mock resuscitations for persons with no resuscitation training. PMID- 10829376 TI - Use of telemedicine in undergraduate teaching of surgery. AB - Telemedicine as an undergraduate teaching method was evaluated by exposing medical students to outpatient videoconferencing. Forty clinical students on a 10 week surgical rotation participated in the study. Telemedicine sessions involved videoconferencing from the general surgical outpatient department located in a sister hospital 8 km away, using a roll-about system connected by ISDN at 384 kbit/s. A questionnaire was used to assess the quality of time spent and educational information obtained by the students. On a scale of 0-10, the students gave an overall rating of 9.7 for outpatient teaching using telemedicine. All 40 students indicated a willingness to return for similar teaching sessions. Even though the results were encouraging, an objective assessment of the knowledge gained by the students is required before the inclusion of this teaching technique in an existing medical curriculum can be recommended unequivocally. PMID- 10829377 TI - Medical communication from emergency scenes using a notepad computer. AB - We have developed a communication system in which data are transferred from the scene of an emergency in the form of an electronic record on a notepad computer by means of a satellite link (MODACOM). The data are received at the dispatch centre and transferred to the admitting hospital automatically. In a prospective study of 53 emergencies the technical suitability of the system was investigated. First-sight information could be fed into the computer in 15 s and transmitted within 3 min of arrival on the scene. For 16 patients with life-threatening conditions, the admitting hospital was notified on average after 13.6 min (SD 6.0), whereas by conventional VHF radio it took 35.5 min (SD 8.9). In addition, more precise information about the patient's condition was received at the hospital. PMID- 10829378 TI - Scoring written material from the Mini-Mental State Examination: a comparison of face-to-face, fax and video-linked scoring. AB - We assessed the reliability of scoring written items of the Mini-Mental State Examination (MMSE) using different communication modes: face to face, fax and videoconferencing. A total of 99 MMSEs were recovered from the records of a community mental health team for the elderly. The written parts of the examination (sentence and pentagrams) were scored--in person, using a faxed copy or over the video-link--according to published criteria. Relative to in-person scoring, sentences could be scored reliably when faxed (kappa = 0.80), as could pentagrams (kappa = 0.71). Sentences could be reliably scored over the videoconferencing link (kappa = 0.70), but pentagrams could not (kappa = 0.47). Caution is required before accepting the results of scoring such tests over video links. PMID- 10829379 TI - Spontaneous intracerebral haematoma with rapidly deteriorating level of consciousness: treatment by a mobile neurosurgeon. PMID- 10829380 TI - Wireless picture transfer as a tool of primary health care. PMID- 10829382 TI - Are bioengineers trend setters or followers? PMID- 10829381 TI - Improving the interface between medical care, telecommunication and legal regulation. PMID- 10829383 TI - Automatic de-noising of knee-joint vibration signals using adaptive time frequency representations. AB - A novel de-noising method for improving the signal-to-noise ratio of knee-joint vibration signals (also known as vibro-arthrographic (VAG) signals) is proposed. The de-noising methods considered are based on signal decomposition techniques, such as wavelets, wavelet packets and the matching pursuit (MP) method. Performance evaluation with synthetic signals simulated with the characteristics expected of VAG signals indicates good de-noising results with the MP method. Statistical pattern classification of non-stationary signal features extracted from time-frequency distributions of 37 (19 normal and 18 abnormal) MP method-de noised VAG signals shows a sensitivity of 83.3%, a specificity of 84.2% and an overall accuracy of 83.8%. PMID- 10829384 TI - Characterisation of three-dimensional anatomic shapes using principal components: application to the proximal tibia. AB - The objective of the research is to determine if principal component analysis (PCA) provides an efficient method to characterise the normative shape of the proximal tibia. Bone surface data, converted to analytical surface descriptions, are aligned, and an auto-associative memory matrix is generated. A limited subset of the matrix principal components is used to reconstruct the bone surfaces, and the reconstruction error is assessed. Surface reconstructions based on just six (of 1452) principal components have a mean root-mean-square (RMS) reconstruction error of 1.05% of the mean maximum radial distance at the tibial plateau. Surface reconstruction of bones not included in the auto-associative memory matrix have a mean RMS error of 2.90%. The first principal component represents the average shape of the sample population. Addition of subsequent principal components represents the shape variations most prevalent in the sample and can be visualised in a geometrically meaningful manner. PCA offers an efficient method to characterise the normative shape of the proximal tibia with a high degree of dimensionality reduction. PMID- 10829385 TI - A portable instrument for non-invasive monitoring of beat-by-beat cardiovascular haemodynamic parameters based on the volume-compensation and electrical admittance method. AB - A new, portable instrument has been developed for simultaneous monitoring of blood pressure, cardiac output and other cardiovascular haemodynamic variables. The measurements are based on the volume-compensation method and the transthoracic electrical admittance method. The instrument is small and light and can be carried by the subject. The portable unit controls the measurement procedures, performs the blood pressure and cardiac output measurement, processes signals and stores almost 32,000 beats of time-series data in a fully automated manner. A conventional personal computer is used to initialize the measurement system, and to reproduce and evaluate the measurement data. The measurement system provides a means to evaluate in detail, without restriction, the subject's cardiovascular haemodynamic responses to daily physical activities as well as to various psycho-physiological stresses. The paper describes a new, portable, cardiovascular haemodynamic instrument and in-the-field test results. Twelve subjects are monitored for trials, 60-210 min, and fewer than 3% of the data in each trial are artifactual. Artefacts are due mainly to body movements. PMID- 10829386 TI - Classification of breast tissue by electrical impedance spectroscopy. AB - Electrical impedance spectroscopy is a minimally invasive technique that has clear advantages for living tissue characterisation owing to its low cost and ease of use. The present paper describes how this technique can be applied to breast tissue classification and breast cancer detection. Statistical analysis is used to derive a set of rules based on features extracted from the graphical representation of electrical impedance spectra. These rules are used hierarchically to discriminate several classes of breast tissue. Results of statistical classification obtained from a data set of 106 cases representing six classes of excised breast tissue show an overall classification efficiency of approximately 92% with carcinoma discrimination > 86%. PMID- 10829387 TI - Analysis of thermal properties of wheelchair cushions with thermography. AB - Thermal properties of wheelchair cushions have been traditionally studied with thermistor probes, which provide temperature values of limited areas (spot analysis). In this paper, we describe a novel procedure based on thermography for assessing the distribution of temperature over the entire surface of wheelchair cushions. The thermal transient during contact with the body (heating phase) and after use (cooling phase) is considered. The procedure was tested in four different seat cushions (with a gel pad, air-filled cells, gel-filled bubbles and foam-filled bubbles) used by a normal subject. Observed results were compatible with the predicted outcomes based on an analysis of the materials and structures. Specifically: (i) air-filled cushions exhibited the fastest thermal transients, gel cushions the slowest transients, while cushions with a mixed structure exhibited intermediate behaviour; (ii) cushions made from flat surfaces of foam exhibited the highest peak temperatures (30.8 degrees C) as compared to those with air-filled cells (30.35 degrees C) or bubble-shaped surfaces (29.7 degrees C); (iii) the average temperature under the thighs was significantly higher than that under the ischiatic area in all cushions (29.6 degrees C compared with 28.7 degrees C, p < 0.05). It is shown that the present method can be used to differentiate between different cushions. Although the 'macro-analysis' inherent in thermography appears to be suited for improving cushion design, this approach should be further investigated to determine its reliability. PMID- 10829388 TI - Artifact reduction in electrogastrogram based on empirical mode decomposition method. AB - Severe contamination of the gastric signal in electrogastrogram (EGG) analysis by respiratory, motion, cardiac artifacts, and possible myoelectrical activity from other organs, poses a major challenge to EGG interpretation and analysis. A generally applicable method for removing a variety of artifacts from EGG recordings is proposed based on the empirical mode decomposition (EMD) method. This decomposition technique is adaptive, and appears to be uniquely suitable for nonlinear, non-stationary data analysis. The results show that this method, combined with instantaneous frequency analysis, effectively separate, identify and remove contamination from a wide variety of artifactual sources in EGG recordings. PMID- 10829389 TI - Detection and analysis of gastrointestinal sounds in normal and small bowel obstructed rats. AB - This study is aimed at detecting gastrointestinal sounds (GIS) and correlating their characteristics with gastrointestinal (GI) conditions. The central hypotheses are that GIS generation depends on the motility patterns and the mechanical properties of the gut, and that changes in those result in measurable differences in GIS. An animal model which included both healthy rats and those with small bowel obstruction (SBO) was developed. The acoustic bursts of GIS were detected by amplitude thresholding the signal envelope. Three methods of envelope estimation were proposed and evaluated. Envelope estimation using a Hilbert transform was found to produce the best results in the current application. The duration and dominant frequency of each detected GIS event was estimated and clear differences between healthy and diseased rats were discovered. In the control state, GIS events were found to consistently be of relatively short duration (3-65 ms). Although the majority of events in the SBO state had similar short duration, infrequent longer events were also detected and appeared to be pathognomonic. Long duration events (> 100 ms) occurred in each of seven obstructed, but in none of 14 non-obstructed, cases (p < 0.001). It is concluded that GIS analysis may prove useful in the non-invasive, rapid, and accurate diagnosis of SBO. PMID- 10829390 TI - Prototype algorithm for automated determination of gastric slow wave characteristics. AB - An algorithm for determining the frequency and propagation time of the gastric slow wave has been designed for integration into a demand gastric pacing system. The algorithm analyses the serosal activity in both the time and frequency domains, and the results are compared to produce a conclusion only when the values are within 5% of each other. Thus, the probability of inappropriate intervention is reduced, at the expense of unidentified segments. The system is verified by comparing the conclusions produced by the algorithm with conclusions from hand analysis of seven canine and one human serosal recordings. The algorithm correctly identifies the slow-wave frequency in the distal portion of the stomach for 90% of the segments, while producing no incorrect results. Slow wave propagation times in the antrum are correctly identified for 84% of the segments, with no incorrect identifications. PMID- 10829391 TI - Real-time brain-computer interfacing: a preliminary study using Bayesian learning. AB - Preliminary results from real-time 'brain-computer interface' experiments are presented. The analysis is based on autoregressive modelling of a single EEG channel coupled with classification and temporal smoothing under a Bayesian paradigm. It is shown that uncertainty in decisions is taken into account under such a formalism and that this may be used to reject uncertain samples, thus dramatically improving system performance. Using the strictest rejection method, a classification performance of 86.5 +/- 6.9% is achieved over a set of seven subjects in two-way cursor movement experiments. PMID- 10829392 TI - Selecting relevant electrode positions for classification tasks based on the electro-encephalogram. AB - The aim is to describe a general approach to determining important electrode positions when measured electro-encephalogram signals are used for classification. The approach is exemplified in the frame of the brain-computer interface, which crucially depends on the classification of different brain states. To classify two brain states, e.g. planning of movement of right and left index fingers, three different approaches are compared: classification using a physiologically motivated set of four electrodes, a set determined by principal component analysis and electrodes determined by spatial pattern analysis. Spatial pattern analysis enhances the classification rate significantly from 61.3 +/- 1.8% (with four electrodes) to 71.8 +/- 1.4%, whereas the classification rate using principal component analysis is significantly lower (65.2 +/- 1.4%). Most of the 61 electrodes used have no influence on the classification rate, so that, in future experiments, the setup can be simplified drastically to six to eight electrodes without loss of information. PMID- 10829393 TI - Computerised evaluation of cognitive and motor function. AB - In this paper, we present a clinical study of computerised tracking in the evaluation of cognitive and motor function. We investigate its use in the assessment of effectiveness of antiepileptic drugs (AEDs) as well as in the process of following the progress of Alzheimer's disease (AD). To simplify the experiments, we introduce real-time adaptation of the target speed. In the study with epileptic patients, three result groups are compared: blood levels of AEDs, scores on standard neuropsychological tests, and scores on computerised tracking and reaction time tests. It is found that the computerised tests are repeatable, reliable and sensitive and may therefore be useful in the evaluation of epilepsy treatment. For example, while the blood levels associated with AEDs lie in the therapeutic range, variations in the optimal speed (OS) between 0.9 and 1.1 (expressed in relative units) are recorded. To significantly simplify the protocol for AD patients while preserving its main features, we introduce signal processing techniques into the data analysis. Local signal property characteristics for AD are found which indicate that the preview tracking of an AD patient is similar to the non-preview tracking of a healthy control. This result is expected since the working memory, which is involved in movement planning, is impaired in AD. In non-preview tracking, healthy control subjects are mostly in tracking mode 1 and have a mean mode duration of 600 ms. In preview tracking, AD patients are mostly in mode 2 with a mean mode duration of 600 ms. PMID- 10829394 TI - Two-point vibrotactile discrimination related to parameters of pulse burst stimulus. AB - Tactile spatial resolution is an important factor in the design of vibrotactile arrays. The two-point discrimination distance is used as a measure of tactile spatial resolution. An experimental study is presented showing the effect of pulse burst stimulus parameters, pulse repetition period and duty cycle on two point vibrotactile spatial discrimination. An array of piezoceramic vibrators is used to measure two-point spatial discrimination on the index finger. In a group of 14 subjects, the average two-point discrimination distance for a pulse repetition period of 1/25s is 2.1 mm (SD = 1.0), whereas for 1/500 s it is 5.1 mm (SD = 0.9). Differences in discrimination distances are statistically significant according to the ANOVA analysis (p < 0.001). Results show that the two-point discrimination distance is better for longer pulse repetition periods. Therefore the pulse repetition period in an excitatory waveform composed of bursts of pulses is important for tactile resolution. No statistically significant differences in discrimination distances are found between bursts of pulses of 50% duty cycle and those of lower duty cycle. The latter result indicates that, by choosing low-duty cycle waveforms for vibrotactile stimulation, the power can be reduced with no loss in two-point discrimination capacity. PMID- 10829395 TI - Simplified model of laser Doppler signals during reactive hyperaemia. AB - Laser Doppler flowmetry (LDF) is a non-invasive method to measure tissue blood flow. During reactive hyperaemia, the LDF signal increases to a peak and then returns to a resting value. A simplified model is developed to explain these variations. The emphasis is on simulating the effects occurring rather than on trying to mimic the anatomical structure of the microcirculation. A single blood vessel is therefore analysed. The increasing value of blood velocity is studied, and vasodilatation as well as vasoconstriction are taken into account. The model parameters are calculated using wavelets. For a 2-min occlusion on a healthy subject, the radius of the vessel is initially 15 microns, increasing to 24.6 microns at the peak, reached 14 s after the release of the occlusion. The model shows that the high value of the LDF signal during the initial phase of reactive hyperaemia is produced by an increasing number of erythrocytes in a cross section, due to vasodilatation rather than an increase in moving blood cell velocities. Moreover, the rapidity of the vasodilatation and vasoconstriction effects determine the rapidity of the signal variations. The paper aims to give a basic solution to develop a numerical model. PMID- 10829396 TI - Cardiac vulnerability assessment from electrical microvariability of high resolution electrocardiogram. AB - Patients susceptible to malignant arrhythmias often have an increased beat-to beat variation of the T-wave of the electrocardiogram. Variability analysis of the T-wave is increasingly used for non-invasive risk assessment. The aim of this study is to evaluate intra-QRS beat-to-beat signal variation and to compare it to ST-T variation. The beat-to-beat, microvolt variation of the QRS and the ST-T segment from 44 patients with coronary heart disease at high risk of suffering from malignant arrhythmias and from 51 healthy volunteers are compared. Variation analysis is carried out on 250 consecutive sinus beats from high-resolution electrocardiograms. The individual beats are filtered using a waveform independent, cubic spline-filter. A variability index of the QRS and ST-T segments is calculated as the integrated standard deviation of corresponding samples inside the area of interest. Patients at risk of suffering from malignant arrhythmias have a significantly higher variability index of both the QRS (median 44.5 ms against 34.7 ms, p < 0.001) and the ST-T segment (median 20.5 ms against 9.8 ms, p < 0.001) compared to the group of healthy subjects. The discriminative ability of the odds variability indices of the QRS and ST-T segments are not statistically different, the ratios being 7.8 (QRS) and 12.6 (ST-T). We conclude that patients at high risk of suffering from malignant arrhythmias are characterised by an increased beat-to-beat microvolt variation of both the QRS and the ST-T segment. Further studies are necessary to evaluate the prognostic potential of depolarisation variability. PMID- 10829397 TI - Can vortices in the flow across mechanical heart valves contribute to cavitation? AB - Cavitation in mechanical heart valves is traditionally attributed to the hammer effect and to squeeze and clearance flow occurring at the moment of valve closure. In the present study, an additional factor is considered--the contribution of vortex flow. Using a computational fluid dynamics analysis of a 2D model of a tilting disk mitral valve, we demonstrate that vortices may form in the vicinity of the inflow side of the valve. These vortices roll up from shear layers emanating from the valve tips during regurgitation. A significant decrease in the pressure at the centre of the vortices is found. The contribution of the vortex to the total pressure drop at the instant of closure is of the order of 70 mmHg. Adding this figure to the other pressure drop sources that reach 670 mmHg, it might be that this is the deciding factor that causes the drop in blood pressure below vapour pressure. The total pressure drop near the upper tip (750 mmHg) is larger than near the lower tip (670 mmHg), indicating a preferential location for cavitation inception, in agreement with existing experimental findings. PMID- 10829398 TI - Validation of automated oscillometric sphygmomanometer (HDBPM) for arterial pressure measurement during haemodialysis. AB - An HDBPM oscillometric sphygmomanometer used for the automatic measurement of arterial blood pressure is evaluated according to the ANSI/AAMI SP10-1992 standard. The accuracy of the HDBPM is ascertained by comparing it against the standard Riva-Rocci ascultatory method. Following the ascultatory method, two independent observers use the HDBPM devise to simultaneously measure the arterial blood pressure in 92 subjects of varying ages and having different blood pressures and arm sizes. High agreement is found when comparing the observers' pressure determinations (within 10 mmHg for 100% of observations). Correlation between the average of two ascultatory determinations and the HDBPM is high both for the systolic (r = 0.98) and diastolic (r = 0.94) pressures. The mean of the differences between the pressures measured by the observers and the HDBPM device are 0.2 mmHg (systolic) and -0.4 mmHg (diastolic). The percentages of readings within 10 mmHg between those taken by the observers and those taken by the HDBPM are 88% (systolic) and 97% (diastolic). These results largely satisfy current requirements. PMID- 10829399 TI - Effect of generator nonlinearities on the accuracy of respiratory impedance measurements by forced oscillation. AB - Measurements of respiratory impedance by means of the forced oscillation technique (FOT) are usually made using a loudspeaker as the excitation device. Its nonlinear nature can introduce artifacts that coincide with the frequencies applied to excite the respiratory system, limiting the accuracy of the impedance estimation. In this paper, this hypothesis is evaluated in the case of both a traditional estimator and the unbiased estimator proposed by Daroczy and Hantos (1982). A simulated study under apnoea conditions in the pressure range 0.5-3.0 cmH2O peak-to-peak reveals that loudspeaker nonlinearities introduce a characteristic pattern of dispersion in both the resistance and reactance curves that can be significantly decreased (p approximately equal to 0.03, signtest) by reducing the nonlinearities. A simulation of spontaneous breathing shows the same pattern, and is observed in the case of traditional as well as unbiased estimators. The dispersion is quantified by the mean absolute distance between the theoretical and simulated data and decreases with the reduction of nonlinearities when impedance is estimated with a traditional estimator (from 6.63 to 4.72% in real estimates and from 6.78 to 3.47% in imaginary estimates) as well as with an unbiased estimator (real estimates from 4.84 to 1.57% and 5.61 to 2.06% in imaginary estimates). Studies with normal subjects show the same dispersion pattern, which decreases if the generator nonlinearities are reduced. These results supply substantial evidence that reducing generator nonlinearities can contribute to the production of more reliable mechanical impedance FOT measurements. PMID- 10829400 TI - Estimating germinability of Plasmopara viticola oospores by means of neural networks. AB - Neural networks are trained to estimate the germination percentages of Plasmopara viticola oospores, overwintered in natural conditions in two viticultural areas in northern Italy, by using climatic (temperature and rainfall) data, as well as the previous germination measurement, as input variables. The 288 available patterns consist of a set of selected independent variables associated with the corresponding germination percentage. All 12 networks investigated converge to a non-linear relationship between the selected independent variables and oospore germination. The highest correlation coefficient (equal to 0.83) between the real and estimated germination percentages is obtained by considering, as input to the network, the climatic data (both temperature and rainfall) recorded during the 40 days before sampling and the germination percentage assessed in the germination assay carried out immediately before the present sampling. PMID- 10829401 TI - Biological-to-electronic interface with pores of ATP synthase subunit C in silicon nitride barrier. AB - An oscillator pore is identified that generates intermittent, large amplitude, ionic current in the plasma membrane. The pore is thought to be composed of 10-12 units of subunit c of ATP synthase. Pore opening and closing is a co-operative process, dependent on the release, or binding, of as many as six calcium ions. This mechanism suggests a more general method of co-operative threshold detection of chemical agents via protein modification, the output being directly amplified in a circuit. Here the authors describe steps in the development of a sensor of chemical agents. The subunit c pore in a lipid bilayer spans a nanometer-scale hole in a silicon nitride barrier. Either side of the barrier are electrolyte solutions and current through the pore is amplified by circuitry. The techniques of laser ablation, electron beam lithography and ion beam milling are used to make successively smaller holes to carry the lipid patch. Holes of diameter as small as 20 nm are engineered in a silicon nitride barrier and protein activity in lipid membranes spanning holes as small as 30 nm in diameter is measured. The signal-to-noise ratio of the ionic current is improved by various measures that reduce the effective capacitance of the barrier. Some limits to scale reduction are discussed. PMID- 10829402 TI - Finite state control of functional electrical stimulation for the rehabilitation of gait. AB - Finite state control is an established technique for the implementation of intention detection and activity co-ordination levels of hierarchical control in neural prostheses, and has been used for these purposes over the last thirty years. The first finite state controllers (FSC) in the functional electrical stimulation of gait were manually crafted systems, based on observations of the events occurring during the gait cycle. Subsequent systems used machine learning to automatically learn finite state control behaviour directly from human experts. Recently, fuzzy control has been utilised as an extension of finite state control, resulting in improved state detection over standard finite state control systems in some instances. Clinical experience over the last thirty years has been positive, and has shown finite state control to be an effective and intuitive method for the control of functional electrical stimulation (FES) in neural prostheses. However, while finite state controlled neural prostheses are of interest in the research community, they are not widely used outside of this setting. This is largely due to the cumbersome nature of many neural prostheses which utilise externally mounted gait sensors and FES electrodes. FES-based control of movement has been subject to the constraints of artificial sensor and FES actuator technologies. However, continued advances in natural sensors and implanted multi-channel stimulators are broadening the boundaries of artificial control of movement, driving an evolutionary process towards increasingly human like control of FES-based gait rehabilitation systems. PMID- 10829403 TI - Fusion of coregistered cross-modality images using a temporally alternating display method. AB - As an aid to the interpretation of functional images, cross-modality coregistration of functional and anatomical images has grown rapidly. Various ways of easily interpreting and visualising coregistered images have previously been investigated; for their display, an intensity-weighted temporally alternating method is used. For brain images, geometric registration involves the automatic alignment method, using the head scalp boundary extracted from the sinogram of a PET emission scan and a surface-matching algorithm; images of the chest or abdomen are registered semi-automatically using a paired point matching algorithm. For the simultaneous display of geometrically registered images, rapid image switching is applied; both images are written with independent colour scales. The rapidly alternating display of two images, synchronised with monitor scanning, induces the fusion of images in the human visual perception system. The accuracy of registration of PET and MRI images is within 2 mm for two point sets. A resulting image is intensified by weighting the display time and/or controlling the intensity map of each image with the degree of interest. This method may be useful for the interpretation and visualisation of coregistered images. PMID- 10829404 TI - 3D reconstruction method from biplanar radiography using non-stereocorresponding points and elastic deformable meshes. AB - Standard 3D reconstruction of bones using stereoradiography is limited by the number of anatomical landmarks visible in more than one projection. The proposed technique enables the 3D reconstruction of additional landmarks that can be identified in only one of the radiographs. The principle of this method is the deformation of an elastic object that respects stereocorresponding and non stereocorresponding observations available in different projections. This technique is based on the principle that any non-stereocorresponding point belongs to a line joining the X-ray source and the projection of the point in one view. The aim is to determine the 3D position of these points on their line of projection when submitted to geometrical and topological constraints. This technique is used to obtain the 3D geometry of 18 cadaveric upper cervical vertebrae. The reconstructed geometry obtained is compared with direct measurements using a magnetic digitiser. The order of precision determined with the point-to-surface distance between the reconstruction obtained with that technique and reference measurements is about 1 mm, depending on the vertebrae studied. Comparison results indicate that the obtained reconstruction is close to the actual vertebral geometry. This method can therefore be proposed to obtain the 3D geometry of vertebrae. PMID- 10829405 TI - Progressive cross-section display of 3D medical images. AB - The paper presents a hierarchical coding algorithm for 3D medical images based upon hierarchical interpolation with radial basis function networks. By using the properties of the Kronecker product, the computation of the network parameters and the 3D image reconstruction are efficiently done in (L4) computation time and O(L3) storage space, when applied to 3D images of size (L x L x L). A further reduction in processing time is accomplished by using sparse matrix techniques. The salient features of the proposed coding method are that arbitrary cross section images can be progressively displayed without reconstruction of the whole 3D image; the first image reconstruction starts as soon as the first data transmission has been completed; no expanding procedure is required in 3D image reconstruction, and the blocking effects are not apparent even in the lowest resolution image. Experimental results using two 3D MRI images, of size (128 x 18 x 64) and with 8-bit grey levels, show that the coding performance is better than that of the 3D DCT coding by about 0.25 bits pixel-1 at higher bit rates, and that the new cross-section display method synthesises the coarsest (finest) section image about six (three) times faster than the standard method that requires the whole 3D image reconstruction. PMID- 10829406 TI - Quantitative assessment of ultrasound-induced resistance change in saline solution. AB - The purpose of the study was the experimental assessment of the interaction coefficient characterising the influence of pressure on the conductivity of electrolyte solutions. Pressure pulses were applied to samples of 9 gl-1 sodium chloride contained in cuboid measurement cells of identical cross-sectional dimensions but different thickness along the acoustic beam axis. The magnitude of the changes induced in cell resistance was recorded for three values of applied pressure increment (delta P = 0.94, 1.39 and 1.78 MPa) and three values of cell thickness (e = 0.58, 1.13 and 1.62 mm). A thick, focused transducer generated short (0.1 microsecond), unipolar pressure pulses. A model accounting for the characteristics of the pressure pulse and the geometry of the measurement cell was developed to predict the ultrasound-dependent changes in the measured electrical resistance. Despite some discrepancy between theoretical and experimental results, discussed in the paper, the results validated the order of magnitude of the interaction coefficient (10(-9) Pa-1). The predictions varied from about 50% (e = 1.62 mm, delta P = 0.94 MPa) to 77% (e = 0.58 mm, delta P = 1.78 MPa) of the experimental values. PMID- 10829407 TI - Integrated pressure-force-kinematics measuring system for the characterisation of plantar foot loading during locomotion. AB - Plantar pressure, ground reaction force and body-segment kinematics measurements are largely used in gait analysis to characterise normal and abnormal function of the human foot. The combination of all these data together provides a more exhaustive, detailed and accurate view of foot loading during activities than traditional measurement systems alone do. A prototype system is presented that integrates a pressure platform, a force platform and a 3D anatomical tracking system to acquire combined information about foot function and loading. A stereophotogrammetric system and an anatomically based protocol for foot segment kinematics is included in a previously devised piezo-dynamometric system that combines pressure and force measurements. Experimental validation tests are carried out to check for both spatial and time synchronisation. Misalignment of the three systems is found to be within 6.0, 5.0 and 1.5 mm for the stereophotogrammetric system, force platform and pressure platform, respectively. The combination of position and pressure data allows for a more accurate selection of plantar foot subareas on the footprint. Measurements are also taken on five healthy volunteers during level walking to verify the feasibility of the overall experimental protocol. Four main subareas are defined and identified, and the relevant vertical and shear force data are computed. The integrated system is effective when there is a need for loading measurements in specific plantar foot subareas. This is attractive both in clinical assessment and in biomechanics research. PMID- 10829408 TI - Improvement of walking speed prediction by accelerometry and altimetry, validated by satellite positioning. AB - Activity monitors based on accelerometry are used to predict the speed and energy cost of walking at 0% slope, but not at other inclinations. Parallel measurements of body accelerations and altitude variation were studied to determine whether walking speed prediction could be improved. Fourteen subjects walked twice along a 1.3 km circuit with substantial slope variations (-17% to +17%). The parameters recorded were body acceleration using a uni-axial accelerometer, altitude variation using differential barometry, and walking speed using satellite positioning (DGPS). Linear regressions were calculated between acceleration and walking speed, and between acceleration/altitude and walking speed. These predictive models, calculated using the data from the first circuit run, were used to predict speed during the second circuit. Finally the predicted velocity was compared with the measured one. The result was that acceleration alone failed to predict speed (mean r = 0.4). Adding altitude variation improved the prediction (mean r = 0.7). With regard to the altitude/acceleration-speed relationship, substantial inter-individual variation was found. It is concluded that accelerometry, combined with altitude measurement, can assess position variations of humans provided inter-individual variation is taken into account. It is also confirmed that DGPS can be used for outdoor walking speed measurements, opening up new perspectives in the field of biomechanics. PMID- 10829409 TI - Intragastric movement assessment by measuring magnetic field decay of magnetised tracer particles in a solid meal. AB - The aim of the study is to evaluate and further develop a non-invasive method for the measurement of intragastric movements. Ferrimagnetic particles (gamma-Fe2O3) are ingested within a standard test meal (pancakes), and given a permanent magnetisation by an applied homogeneous magnetic field. Magnetic measurements are performed outside the stomach, using fluxgate magnetometers anterior and posterior, in seven periods of 12 minutes each. Any movements caused by grinding and mixing in the ingesta would result in a misalignment of the magnetic moments, and hence in a decay of the external magnetic field. Twelve healthy male volunteers, aged 26.7 +/- 2.7 years (mean +/- SD), are studied. After an initial phase of low and almost linear flux density decay (32 +/- 19%, from minute 10 to minute 22 postprandially), the mean rotation and decay gradually increases (67 +/ 21%, from minute 100 to 112), and the decay becomes bi-exponential. It is hypothesised that the bi-exponential nature of the decay corresponds to the relative fractions, solid and semifluid, of gastric content. The decay curves often show superimposed 3 cycles min-1 variations, which correlate well with the basic electrical rhythm of the stomach measured by electrogastrography (EGG). It is concluded that these variations are induced by gastric contractions. PMID- 10829410 TI - Silicon-based microelectrodes for neurophysiology, micromachined from silicon-on insulator wafers. AB - A process is described for the fabrication of silicon-based microelectrodes for neurophysiology using bonded and etched-back silicon-on-insulator (BESOI) wafers. The probe shapes are defined without high levels of boron doping in the silicon; this is considered as a step towards producing probes with active electronics integrated directly beneath the electrodes. Gold electrodes, of 4 microns by 4 microns to 50 microns by 50 microns are fabricated on shanks (cantilever beams) 6 microns thick and which taper to an area approximately 100 microns wide and 200 microns long, which are inserted into the tissue under investigation. The passive probes fabricated have been successfully employed to make acute recordings from locust peripheral nerve. PMID- 10829411 TI - Information domain analysis of cardiovascular variability signals: evaluation of regularity, synchronisation and co-ordination. AB - A unifying general approach to measure regularity, synchronisation and co ordination is proposed. This approach is based on conditional entropy and is specifically designed to deal with a small amount of data (a few hundred samples). Quantitative and reliable indexes of regularity, synchronisation and co ordination (ranging from 0 to 1) are derived in a domain (i.e. the information domain) different from time and frequency domains. The method is applied to evaluate regularity, synchronisation and co-ordination among cardiovascular beat to-beat variability signals during sympathetic activation induced by head-up tilt (T), during the perturbing action produced by controlled respiration at 10, 15 and 20 breaths/min (CR10, CR15 and CR20), and after peripheral muscarinic blockade provoked by the administration of low and high doses of atropine (LD and HD). It is found that: (1) regularity of the RR interval series is around 0.209; (2) this increases during T, CR10 and HD; (3) the systolic arterial pressure (SAP) series is more regular (0.406) and its regularity is not affected by the specified experimental conditions; (4) the muscle sympathetic (MS) series is a complex signal (0.093) and its regularity is not influenced by HD and LD; (5) the RR interval and SAP series are significantly, though weakly, synchronised (0.093) and their coupling increases during T, CR10 and CR15; (6) the RR interval and respiration are coupled (0.152) and their coupling increases during CR10; (7) SAP and respiration are significantly synchronised (0.108) and synchronisation increases during CR10; (8) MS and respiration are uncoupled and become coupled (0.119) after HD; (9) the RR interval, SAP and respiration are significantly co ordinated (0.118) and co-ordination increases during CR10 and CR15; (10) during HD the co-ordination among SAP, MS and the respiratory signal is larger than that among the RR interval, SAP, MS and the respiratory signal, thus indicating that the RR interval contributes towards reducing co-ordination. PMID- 10829412 TI - Arterial baroreflex influence on heart rate variability: a mathematical model based analysis. AB - The influence of the arterial baroreflex on the heart rate variability is analysed by using a mathematical model of heart rate baroreceptor control. The basic mechanisms of the model, sufficient to elicit heart rate variability include: systemic circulation, a non-pulsatile cardiac pump and nonlinear negative feedback simulating arterial baroreflex closed-loop control of the heart rate (-3 bpm/mmHg as maximum reflex sensitivity). The latter reproduces, through two distinct delayed branches (0.8 and 2.8 s), the short-term autonomic control effected respectively by sympathetic and parasympathetic divisions on the sinus node. By means of this model, two distinct self-sustained oscillatory components with incommensurate frequencies (0.1 and 0.26 Hz) are reproduced. Frequencies of these two oscillatory components closely agree with the main heart rate rhythms in humans (0.09 +/- 0.01 Hz and 0.26 +/- 0.01 Hz). When sympathetic-mediated regulation prevails over parasympathetic activity, simulated heart rate oscillation is characterised by a low frequency (approximately 0.1 Hz). On the other hand, a high-frequency oscillatory component (approximately 0.26 Hz) appears when enhanced vagal activation or partial inhibition of the sympathetic control is simulated. When both autonomic divisions are operative, both low- and high-frequency components are present and the heart rate oscillates quasi periodically. This variability in heart rate at different frequencies is reproduced without including outside perturbations and is due to the nonlinear delayed structure of the closed-loop control. Bifurcation theory of nonlinear system is used to explain the high sensitivity of the heart rate oscillatory pattern to model parameter changes. PMID- 10829413 TI - Computerised acoustical respiratory phase detection without airflow measurement. AB - A simple, non-invasive acoustical method is developed to detect respiratory phases in relationship to swallows without the direct measurement of airflow. In 21 healthy subjects (4-51 years) breath sounds are recorded at the trachea and at five different recording locations at the chest wall, with simultaneous recording of airflow by a pneumotachograph. The chest signal with the greatest inspiratory expiratory power difference ('best location') is either in the mid-clavicular line in the second interspace on the left or third interspace on the right. Using the 'best location' on the chest wall and the tracheal signal, a phase detection algorithm is developed and achieves 100% accuracy in the estimation of respiratory phases without using the measured airflow signal. Thus, acoustically monitoring breaths and swallows holds promise as a non-invasive and reliable assessment tool in the study of swallowing dysfunction. PMID- 10829414 TI - From the Tissue Engineering and Biomaterials Symposium of the XIe Entretiens du Centre Jacques Cartier held in Lyon in December 1998. PMID- 10829415 TI - Comparative analysis of different collagen-based biomaterials as scaffolds for long-term culture of human fibroblasts. AB - Biodegradable scaffolds, along with cells, are important components of most tissue-engineered constructs. In the study, there is a comparison of the behaviour of human fibroblasts cultured for up to six weeks in four different collagen-based three-dimensional matrices, in the form of sponges composed of pure native type I collagen (control), of collagen-GAG-chitosan (CGC) and of collagen cross-linked by two concentrations of diphenylphosphorylazide (DPPA-2 and DPPA-3). Variations in size and weight of the sponges, as well as fibroblast growth and migration, and total protein and collagen synthesis, are determined with time in culture. Owing to their low thermal stability, the partial denaturation and dissolution of the control sponges after incubation at 37 degrees C lead to considerable contraction and low cell proliferation. CGC sponges, stabilised by ionic interactions between the different components, show, after six weeks, limited contraction (20%) and weight increase (10% when seeded) and high cell growth (threefold increase). Similar results are obtained with weakly, cross-linked (DPPA-2) collagen sponges. Highly cross-linked (DPPA-3) sponges do not contract, whereas weight gain and cell proliferation are no different from those found with CGC and DPPA-2 sponges. Similar levels of total protein and collagen synthesis are shown for fibroblasts seeded in different matrices, with a slight general decrease (twofold) after three weeks, a much lower value than that observed with fibroblasts in culture within a contracted collagen gel (sixfold). Furthermore, the fraction of neo-synthesised collagen deposited in the sponges after six weeks represents more than 60% of the total, compared with only 10% obtained with fibroblasts in monolayer culture or 30% within a collagen gel. These results indicate that the matrices, particularly the CGC and DPPA-2 sponges, provide excellent supports for fibroblast growth and the formation of dermal and skin equivalents. PMID- 10829417 TI - Alternative blood conduits: assessment of whether the porosity of synthetic prostheses is the key to long-term biofunctionality. AB - The paper examines the effects of water permeability on solid particle (platelet) adhesion and lipid transport through the wall of a blood conduit. Also tested is the capacity of external supports to reduce lipid infiltration into venous grafts. The results indicate that water permeability not only facilitates particle adhesion, but also affects the spatial distribution of the adhesion. The presence of filtration flow leads to a concentration polarisation of atherogenic lipids at the blood/wall interface, with increased lipid concentration from the bulk value towards the interface, thus enhancing the drive potential for lipid infiltration into the vessel wall. An external support to a venous graft guards against excessive distention and significantly reduces lipid infiltration into the venous wall. These results strongly suggest that too high a water permeability or porosity can lead to the late failure of arterial grafting by affecting blood cell interaction with the graft and lipid infiltration into the wall. Therefore the pore structure of an arterial prosthesis is crucial to its long-term biofunctionality. Ideally, a synthetic prosthesis should display pores of adequate size and a structural network that promotes tissue ingrowth, while maintaining water porosity at a physiological level. PMID- 10829416 TI - Collagen-based biomaterials as 3D scaffold for cell cultures: applications for tissue engineering and gene therapy. AB - Many substances are used in the production of biomaterials: metals (titanium), ceramics (alumina), synthetic polymers (polyurethanes, silicones, polyglycolic acid (PGA), polylactic acid (PLA), copolymers of lactic and glycolic acids (PLGA), polyanhydrides, polyorthoesters) and natural polymers (chitosan, glycosaminoglycans, collagen). With the rapid development in tissue engineering, these different biomaterials have been used as three-dimensional scaffolds and cell transplant devices. The principal biochemical and biological characteristics of the collagen-based biomaterials are presented, including their interactions with cells (fibroblasts), distinct from those of synthetic polymers, and their potential use in gene therapy through the formation of neo-organs or organoids. PMID- 10829418 TI - Characterisation of elastin and collagen in aortic bioprostheses. AB - Porcine aortic valves used as cardiac valve bioprostheses are well adapted to physiological functions in the short term, but they lack long-term durability. Several multi-step extractions have been performed to obtain a perfectly acellular matrix. A new physical methodology is proposed to evaluate the resulting fibrous protein damage after biochemical extraction (TRI-COL and SDS). Thermal analysis techniques are adapted to collagen and elastin characterisation in the solid state. The aortic tissue thermal transitions are determined by differential scanning calorimetry (DSC): elastin glass transition is observed around 200 degrees C, and collagen denaturation is observed around 230 degrees C. These parameters are characteristic of the elastin network arrangement and of collagen triple-helix stability. The technique of thermostimulated currents (TSC) is well suited to specify the chain dynamics of proteins. The low-temperature relaxations observed in both collagen and elastin are associated with localised motions, whereas the high-temperature modes are attributed to more delocalized motions of the chains. Therefore TSC and DSC spectrometries allow physical parameters specific to collagen and elastin to be obtained and their interaction in aortic tissues to be determined. According to the significant evolution of these parameters on SDS samples, the destabilizing effect of this detergent is highlighted. PMID- 10829420 TI - French legal framework relating to human tissues and cells. AB - Thousands of patients receive human tissue grafts every year. Developments in cell and tissue engineering have also increased considerably the number of available products of human origin. France has very strict regulations, in part stimulated by problems of public health and ethics that have emerged in recent years and also in part as a result of a report by the 'Inspection Generale des Affaires Sociales' on the removal and grafting of human tissues in May 1993. These have resulted in two laws on bio-ethics being passed, in July 1994, that are the basis of current legislation and represent the first steps in differentiating between organs and tissues or cells. Henceforth, the French legal framework covering tissues and cells of human origin has been increased to include a large number of legislative texts and regulations. The fundamental ethical principles that are consent, free donation, anonymity, no publicity and respect for public health have become a major ethical imperative that applies to all products originating from the human body including tissue and cells. In addition, specific provisions have been made covering: removal (conditions for removal and system for authorization); conservation, transformation, distribution, packaging, import and export of tissues and cells; and tissue and cell grafts. Finally, penal and administrative sanctions have been foreseen where there is non-compliance with these regulations. PMID- 10829419 TI - Tissue engineering of the vascular system: from capillaries to larger blood vessels. AB - Tissue engineering is a novel approach to the repair of wounded tissues. Application of this technology to the vascular system is important because of the fundamental nutritional role of the vasculature. This perspective is currently being applied to the first tissue-engineered organ: the skin. Knowledge of capillary constitution and factors inducing their formation has led to attempts to induce their formation in reconstructed skin. Such vascular conduits grown in vitro could also benefit the nutrition of tissues and organs in vivo. The paper reviews recent progress in the in-vitro development of vascularised skin and tissue-engineered blood vessels. It points out the necessity of obtaining pure and well-characterised cultures of the different cell populations that are the basic building blocks of the reconstructions. The importance of an adequate cell culture environment (nutrients and bi- or tri-dimensional scaffolds for cells) for success in elaborating a reconstructed living tissue able to replace the original is emphasised. Engineered tissues can serve not only as tissue replacements but also as in-vitro models for research in organ physiology and physiopathology. These tissues are also attractive vehicles for gene therapy, one of the more promising new methods of disease treatment. PMID- 10829421 TI - Advantages of using a bank of allogenic keratinocytes for the rapid coverage of extensive and deep second-degree burns. AB - In 1975, serial subculture of human keratinocytes was first described. Clinical application of this discovery was made possible after the preparation of these cells into epithelial sheets. In 1981, the earliest application of cultured autologous epithelia was made for the treatment of extensive third-degree burns. Although the most important advantage is the large surface area obtained from a relatively small biopsy of healthy skin from the patient, a disadvantage is the delay, which is too long, especially for the treatment of extensive deep burns. This delay leads to denutrition and infection of the burn wounds, which in turn risks the life of the patient and jeopardizes the engraftment of the cultures. More recently, allogenic cultured epidermis, obtained more quickly from donor skin, has been described in the treatment of leg ulcers, repair of skin donor site harvested for split thickness autograft, dermatological diseases and in second-degree burns, although limited to certain areas. In this last case, grafted cells act by stimulation of epithelialisation from the adnexal appendages. To be able rapidly to treat patients suffering extensive and deep second-degree burns, a bank of allogenic keratinocytes has been created, with due attention to safety and security. The paper demonstrates the advantages of using allogenic keratinocytes in the first phase of treatment of a 97% deep second degree burn patient awaiting autologous cultured keratinocytes. The time required for complete healing achieved using such a strategy is compared with the results obtained after treatment using autologous sheets of two patients burnt on 80% and 82% of their total body area. The treatment of these two latter patients is relatively long and complicated by potentially lethal problems. In the 97% burnt patient, however, the clinical course is shorter and without complication. Moreover, autologous and allogenic cultured epithelia give good aesthetic results, without the mesh aspect obtained with a split-thickness autograft, and also without the discomfort for the patient of removing a sample of skin. Deep second-degree burns are an application of choice for the cultured epithelia, as the presence of the dermis avoids retractions responsible for functional complications usually observed in third-degree burns where dermis is absent. Because of the safety of the bank of allogenic keratinocytes, the treatment of extensive and deep second-degree burns has become safer and faster, with better functional and aesthetic results. PMID- 10829422 TI - An example of political arithmetic: the evaluation of spa therapy at the Georgian Bath Infirmary, 1742-1830. PMID- 10829423 TI - Lives in many hands: the medical landscape in Lancashire, 1700-1820. PMID- 10829424 TI - The key to success: the role of local government in the organization of smallpox vaccination in Sweden. PMID- 10829425 TI - Dracunculiasis in the Caribbean and South America: a contribution to the history of dracunculiasis eradication. PMID- 10829426 TI - The archive of the Queen's Nursing Institute in the Contemporary Medical Archives Centre. PMID- 10829427 TI - Building a research capacity for occupational medicine. PMID- 10829428 TI - Low-level hydrocarbon solvent exposure and neurobehavioural effects. AB - Studies of workers exposed to hydrocarbon solvents are reviewed in order to address the question, 'Does long-term human exposure to hydrocarbon solvents at concentrations around occupational exposure limits result in clinically important neurobehavioural effects?' Studies selected evaluated exposure-response (E-R) trends with some control of potential confounders. Tests of neurobehavioural performance were classified into specific functional categories, within those of cognitive, psychomotor and sensory functions to increase specificity and power to detect patterns of effect. The weight-of-evidence was evaluated with respect to criteria for determining causality. The temporality criterion was met as latency was adequate for the occurrence of chronic effects. There were few significant associations, and when present, were consistently weak even in the most-exposed workers. Exposure response showed no consistent or significant pattern for any tests of functional mortality. The weight of evidence suggests that exposure to hydrocarbon solvents at current limits does not appear to cause adverse neurobehavioural effects. PMID- 10829429 TI - Association between bronchial hyper-reactivity and exposure to silicon carbide. AB - A clinical study of 191 workers in the silicon carbide production industry was carried out in order to determine the prevalence of respiratory symptoms (wheezing, dyspnoea and breathlessness after exercise). Such symptoms occurred in 50 (26.1%) subjects, of whom 24 accepted testing for bronchial reactivity. Of these 24, 19 (79.2%) were positive. These findings suggest that exposure to SiC may be associated with bronchial hyper-reactivity. Positive results were more frequent in women than in men, while differences between smokers and non-smokers were insignificant, a phenomenon that has often been noted in persons exposed to massive risk. PMID- 10829430 TI - Prevalence of low back pain among staff in a rural hospital in Nigeria. AB - A cross-sectional study was carried out in a rural hospital in south-western Nigeria to determine the prevalence of low back pain among its staff. The questionnaire administered to staff sought information on social and demographic characteristics, job history, smoking status, frequency and severity of low back pain and factors predisposing to low back pain. Seventy-four out of a total of 80 workers participated in the study. The prevalence of low back pain among staff was 46%. The highest prevalence of back pain (69%) was recorded among nursing staff, followed by secretaries/administrative staff (55%) and cleaners/aides (47%). Heavy physical work (45%), poor posture (20%) and prolonged standing or sitting (20%) were the most frequent activities reported to be associated with low back pain among these workers. The prevalence of low back pain among these workers is comparable to that of workers in high income countries. Health education on posture and correct lifting techniques can be introduced to reduce the burden of low back pain among these workers. PMID- 10829431 TI - Otorhinolaryngological assessment and psychological adjustment in tanning industry workers. AB - The purpose of the study was to verify the presence of any symptoms and otorhinolaryngological pathologies and to determine any discomfort and psychophysical changes among workers in the tanning industry. The study involved 129 subjects working in tanning industries in the Chiampo valley (Veneto region of Italy). Their clinical history was recorded and they underwent objective ENT examination and pure-tone audiometry. For the psychodiagnostic evaluation, four questionnaires were administered (EPI, STAI, ZUNG and GHQ). ENT assessment revealed no objective problems and findings were within the normal range in the majority of cases. Hearing function was also generally normal or within the normal range for an individual's age. Psychological evaluation revealed a valid psychological adjustment in almost all cases, despite the majority of workers reporting some subjective discomfort attached mainly to the particularly unpleasant smell in a micro-climate with a high rate of humidity. The findings of this preliminary investigation enable us to claim that the working environment in the tanning industries is not severely harmful for the upper airways and hearing function. It is worth noting the normality of the workers' psychological adjustment, which indicates a valid psycho-physical balance in these subjects who operate in an environment with unpleasant features. PMID- 10829432 TI - Lifestyle and serum gamma-glutamyltransferase: a study of middle-aged Japanese men. AB - The association of lifestyle factors with serum gamma-glutamyltransferase (GGT) was examined in 1176 Japanese male office workers aged 40-59 years. Those who had serum aminotransferase levels exceeding the normal range and/or who took prescription medication for, or had a past history of, liver disease were excluded. From the logistic regression analysis, lifestyle factors showing a positive association with high serum GGT (> or = 60 U/l) were body mass index, alcohol intake, cigarette smoking, and snacking between meals. On the other hand, coffee drinking showed an inverse association with high serum GGT. As for the relationships between snacking between meals and other lifestyle factors, snacking between meals was significantly related to alcohol intake (negative) and coffee drinking. Influences of snacking between meals on serum GGT are likely to be indirectly mediated through coping mechanisms of less alcohol drinking and more coffee drinking. Our findings support the conclusions that alcohol drinking, obesity, and cigarette smoking are positively associated with raised serum GGT and that coffee consumption is inversely related to raised serum GGT. PMID- 10829433 TI - A review of current literature on physiological tests and soft tissue biomarkers applicable to work-related upper limb disorders. AB - Work-related upper limb disorders (WRULD) are among the most commonly reported occupational illnesses. Epidemiological evidence of work-relatedness has been reported for a number of conditions, including carpal tunnel syndrome, hand/wrist tendinitis, shoulder tendinitis and hand-arm vibration syndrome. A range of electrodiagnostic techniques and psychophysical tests has been used to assess neurological dysfunction associated with WRULD, whereas only very few studies have examined biochemical markers of soft tissue injury. This report reviews key literature on physiological tests and biochemical markers of musculoskeletal stress/injury, which are applicable to studies of WRULD. The paper concludes by proposing new ways that testing might be implemented during occupational health surveillance to enable early warning of impending problems and to provide more insight into the underlying nature of soft tissue disorders. PMID- 10829434 TI - Exposure measurement in occupation and environmental epidemiology used for quantitative risk assessment. PMID- 10829435 TI - Aspirational targets for occupational health. AB - The new Occupational Health Strategy for Great Britain will have national aspirational targets. These will be common goals that all partners can work to, and should help to inspire local and specific targets for action. More attention needs to be paid to looking at outcomes, so that we can reduce the number of people suffering from work-related ill-health, as well as improving the health of the workforce. Both local and national targets will need evaluating, so that progress can be seen. Organisations who share their success, as well as challenges in making progress will help to spread improving practice. PMID- 10829436 TI - What are the causes of a perforated nasal septum? PMID- 10829437 TI - Auditing health promotion. AB - This review of worksite health promotion (or wellness) programmes in the United States begins with a brief description of the essential elements of effective worksite health promotion programmes citing leading authorities. The evidence supporting the value of worksite health promotion is summarized. The importance of relying on major theories related to health education and health promotion is advocated. Current health promotion trends emerging in the United States are also discussed. PMID- 10829438 TI - Occupational epidemiology. AB - The epidemiological literature for assessing risk in many, if not most, modern occupations has now become sufficiently obsolete that it can no longer be depended upon to guide either prevention or adjudication of compensation. This obsolescence must be dealt with by developing new sources of information pertinent to occupational hazards and the risks associated with most occupations. Ideally, a comprehensive surveillance mechanism that would be automatically updated for the changing risk in a changing economy would be ideal and may be attainable with further developments in health information technology. The characteristics of such a system are described. However, there are many obstacles to such a system which appear insurmountable in the short term. A more eclectic plan for cooperation and data-sharing would help in the short term and would establish a pattern of collaboration that could both place adjudication on a more solid foundation and avoid allegations of collusion in business. The general outline for a practical programme of collaboration along these lines is presented. PMID- 10829439 TI - A review of the healthy worker effect in occupational epidemiology. PMID- 10829440 TI - [Aging and urology]. PMID- 10829441 TI - [Conservative parenchymal surgery in kidney neoplasms]. AB - OBJECTIVES: To analyze retrospectively a series of 60 nephron sparing surgeries (NSS) in patients presenting with renal masses. MATERIAL AND METHODS: The clinical histories of 59 patients undergoing NSS between january 1978 and december 1997 were reviewed. 40 were males, the mean age 54.4 years (range 17-77 years). 10 (17%) had bilateral synchronous tumors, 15 (25%) had a solitary kidney and 3 a renal insufficiency. In 25 patients NSS was obligatory, while in 34 it was elective. 30 (50%) of the renal masses had been diagnosed incidentally. Overall, 49 kidneys underwent an enucleation and 11 a partial nephrectomy. RESULTS: The histopathological exam revealed 43 (71.6%) renal cell carcinomas (RCC), 2 (3.4%) urothelial carcinomas, 6 (10.2%) oncocytomas, 7 (11.6%) angiomyolipomas and 2 (3.4%) multilocular cystic nephromas. Among the RCC, 8 had < or = 3 cm, 22 between 3.1-5 cm, 10 between 5.1-8 cm and 3 more than 8 cm. 31 (72.1%) were well encapsulated and only 2 (4.6%) invaded the nearby parenchyma. 7 (16.2%) were pT1, 34 (79%) pT2 and 2 (4.6%) pT3. 39 (90.3%) were G1 or G2, and only 4 (9.3%) were G3. Two patients (3.3%) died perioperatively, both from the obligatory group. 3 (6.8%) of the patients with malignant tumors progressed and died, all with CCR and from the obligatory surgery group. Another 3 patients of this group developed local recurrences, but were rescued with iterative surgery (conservative in 1); 2 of these patients had sporadic CCR, another a von Hippel Lindau disease. None of the 34 patients who underwent elective NSS progressed, nor recurred locally and all are alive and disease free; only 14 (56%) of the 25 patients operated on by necessity survived, although 6 of them died of causes not related with the kidney tumor. The cancer specific survival at 58.3 months of follow-up (range 5-187 months) is 86.8% for the whole series, 100% for the elective NSS group and 68.7% for the obligatory NSS group. 19 (31.6%) patients developed complications, 14 among the obligatory group; two died (hemorrhage and sepsis). 3 patients presented acute renal failure, but none required dialysis; presently 4 (6.7%) patients have poor renal function and 1 requires dialysis. CONCLUSION: NSS affords good control of renal carcinoma; in the cases operated on electively, the 100% cause specific survival yielded by NSS supports the use of conservative surgery in patients with well delimited tumors and normal contralateral kidney. PMID- 10829443 TI - [Percutaneous treatment of retroperitoneal abscesses]. AB - The retroperitoneal abscess is an uncommon disease, that must be treated by drainage. The progressive use of the percutaneous drainage, under ultrasound or computed tomography guidance (CT), has changed the therapeutical management and has demonstrated to be a valid alternative to surgical drainage. From 1986 to 1998, 16 patients with retroperitoneal abscesses were treated by percutaneous drainage (14 with CT and 2 with ultrasound guidance). This method eradicated the abscess in 13 cases, in 2 was necessary a new function to cure the abscess, and 1 patient, with a severe sepsis, died. Percutaneous drainage was the unique treatment used in 12 patients. In the remaining four, the patients' clinical status improved after percutaneous drainage, and they were able to undergo subsequent elective nephrectomy. CONCLUSIONS: Percutaneous drainage of retroperitoneal abscesses has been established as a viable alternative to surgical intervention. This method can resolve the abscess or improve the patient' clinical status to undergo elective surgery. PMID- 10829442 TI - [Prostatic benign hypertrophy: review of effectiveness, tolerance, and impact on quality of life of prolonged treatment with alfuzosin]. AB - OBJECTIVES: To evaluate the effectiveness of Alfuzosin in terms of long term effect in BHP patients. To analyze the effectiveness according to the observer, the treatment compliance and the tolerance of alfuzosin, and specifically, cardiovascular and sexually adverse events. Also, to define the clinical profile of the BPH patients that spontaneously seek medical treatment in Spain. DESIGN OF THE STUDY: Observational, descriptive, multicentric and 6-month retrospective study. DATA SOURCES AND COLLECTION: From the patient reports of BPH patients treated with alfuzosin during more than 6 months, data gathered were, along with usually anamnesic data and physical exploration (age, symptoms, blood pressure, e.g.), those related to treatment response, effectiveness, compliance and tolerance of Alfuzosin in the first and second quarter. Data of the 1635 patients initially recruited were considered to define the clinical profile of this population. Effectiveness and tolerance analysis were undertaken with the 911 patients who followed the treatment during the 6-month study. RESULTS: After 6 months of treatment an improvement of the mean intensity of each of the urinary symptoms were statistically significant (p < 0.005). Also, the mean score of each of the analyzed variables and of the I-PSS global score were reduced significantly (p < 0.005). After 6 months of treatment the effectiveness evaluated by the observer was good or very good for the 88.5% of the patients. Treatment compliance was very good; from the 911 patients finishing the study, 96.4% continued with the initially prescribed treatment. Just 16 patients (3%) developed adverse events; measures undertaken were to modify the dosage or to interrupt the treatment (2 cases). The specific study of cardiovascular adverse events showed a mild reduction in the systolic (142 mmHg versus 137.86 mmHg) and diastolic blood pressure (83.53 mmHg) versus 80.72 mmHg), and the heart rate (76.80 l/m versus 76.04 l/m). The mean score of sexual life showed an improvement in all the items studied, although very slight. CONCLUSIONS: The usefulness, effectiveness and tolerance of a long term treatment with alfuzosin in HPB patients was confirmed. Alfuzosin is considered a first choice treatment in patients with the clinical profile described. PMID- 10829444 TI - [Significance of simple radiography of the abdomen in nephritic colic]. AB - BASIS: The utility of the abdominal plain film in the emergency department to evaluate urolithiasis is questionable. Between another things, there are influence of the different formation of distinct examinators. METHODS: A retrospective study in 65 patients referrals to urology in 1997 and 1998 by suspicion of renal colic was carried out. The radiography of abdomen took in the emergency room the day of the income is reviewed with a protocol, by a resident of Familiar and Community Medicine and by the urologist, separately and without information of the medical records. RESULTS: We have excluded three patients. At last, we diagnose 49 patients as renal colic with certainty, 6 patients with doubtful diagnosis and 7 patients with distinct diagnosis. The urologist achieved the most adequate reading in 11 cases; in 7 was best the resident, to equal in 31, and in 13 the differences were not important. The resident obtained of radiography a sensitivity of the 67% and a specificity of the 42% and the urologist 71% and 57% respectively. CONCLUSIONS: There are few differences of reading, at least when the resident dispose a protocol of interpretation. Despite to the almost null diagnostic utility of the radiography, supply additional ation, and permit to value the evolution with a few aggressive technique. PMID- 10829445 TI - [Follow up of patients with bladder cancer treated with radical cystectomy]. AB - Radical cystectomy provides excellent local control of the disease in cancer patients, although specific 5-year mortality is about 50%. Additionally, nearly half the patients show complications after surgery usually due to urinary by pass. Patients should undergo periodical monitoring for life. For rescue treatment to be as effective as possible, follow-up schemes should allow for early detection of relapsing-recurrent tumours, metastasis and complications. Knowledge of the "natural evolution" in these patients allows to optimise the frequency of visits and complementary examinations based on each patient's risk at specific time-points. Cost-efficacy relationship will also be optimised. We analyze here the current literature and our own experience of what we know as "natural evolution" of these patients and propose a follow-up scheme for patients with cancer of the bladder treated with radical cystectomy and urinary by-pass. PMID- 10829446 TI - [Mucinous carcinoma of the prostate: critical review of Elbadawi's criteria]. AB - OBJECTIVES: We present our experience on prostatic mucinous adenocarcinoma and at once we practice an actualization and a critical review of Elbadawi's criteria. METHODS: After reviewing 206 prostatic carcinomas diagnosed in our hospital, we describe one case that fulfill criteria for being considered a prostatic mucinous adenocarcinoma. We also carry out a wide literature review trying to define anew the including criteria of this tumour by the light of modern knowledge and technology. CONCLUSIONS: We think that for accepting a tumour as a prostatic mucinous adenocarcinoma, this have to fulfill the following criteria: 1. "More than 25% of a significative tumoral sample is mucinous pattern and present, single or clustered, tumour cells floating in immunohistochemically probed, acidic and neutral, mucin lakes". 2. "Tumour is Gleason 3-4 cribiform pattern with direct transition to colloid areas and usually coexist with classic adenocarcinoma but papillary growth patterns should be excluded. Tumour may contain a moderate proportion of signet-ring cells". 3. "Immunohistochemical staining for PSA have to be strongly positive in both, cribiform and mucinous, areas". 4. "Those PSA nonreactive, or only focally positive, mucinous adenocarcinomas, could be labeled as prostatic only when local or distant mucinous carcinomas are ruled out". PMID- 10829447 TI - [Extracorporeal shock-wave lithotripsy: anxiety and pain perception]. AB - Three factors in extracorporeal shock-wave lithotripsy (ESWL) result in pain: shock wave pressure, size of focal area and pressure distribution at entry in the skin. Our Lithotripsy Unit has performed 21,000 outpatient treatments (Dornier HM 4 Lithotripter) with no sedation or anaesthetics. Pain requiring treatment discontinuation resulted in 10% instances. Anxiety-associated pain increases the perceived intensity of pain and influences the emotional response to pain. OBJECTIVE: To determine whether the variability in the response to the pain stimulus caused by ESWL is related to the patient's status/trait of anxiety. METHOD: Sample 20 patients; inclusion criteria: pyelic lithiasis > 2 cm, no previous SWEL and no obesity. One SWEL session (2500 pulses, 22 Kv). Anxiety was first evaluated using the status/trait of anxiety index (STAI) questionnaire; after treatment, pain was evaluated on a Numerical Analogic Scale. RESULTS: Pain scores ranged from 2 to 8, mean score 4.1 (S.D. = 1.67); mean score for trait of anxiety 21.8 (S.D. = 8.52), and status of anxiety 29 (S.D. = 5.89). A significant correlation was found between pain-anxiety trait (R = .51; p = .02), which was higher when compared to pain-anxiety status (R = .67; p = .001). CONCLUSIONS: SWEL is a painful therapy (95% of sample had mild-to-moderate pain); pain perception is increased by status of anxiety (45% of pain variance) more than by the trait of anxiety (26.3%); therefore, usage of sedation-analgesia in patients with high anxiety status would improve the lithotripter efficacy ratio. PMID- 10829448 TI - [Renal cell carcinoma in patient with situs inversus and Kartagener syndrome]. AB - A case of a adenocarcinoma renal in a patient with situs inversus complete to which was associated bronchiectasis and chronic sinusitis (kartagener's syndrome) is reported. It is the third case described in the international literature of a renal cells carcinoma in a patient with situs inversus totalis and the first in patient the one which has the triada classic of the kartagener's syndrome plus sterility. The tumor was discovered in a way incidental upon accomplishing a TAC toracoabdominal and was solved through nefrectomia for lumbotomy approach. PMID- 10829449 TI - [Synchronous prostatic carcinosarcoma: report of 2 cases and review of the literature]. AB - OBJECTIVE: Carcinosarcoma of the prostate is a biphasic tumor containing adenocarcinoma and recognisable sarcomatous components. It is a rare neoplasm with less than 40 reported cases. We describe two additional examples showing carcinosarcoma in the initial pathologic material (synchronous presentation). METHODS AND RESULTS: The patients were 71 and 78 year-old, respectively. The primary prostatic tumour in case one was composed of adenocarcinoma admixed with a neoplastic mesenchymal component that displayed foci of chondrosarcoma, rhabdomyosarcoma and angiosarcoma. The adenocarcinoma in the second case was admixed with spindle cell sarcoma and chondrosarcoma. Both patients died of disease after surgery, 3 and 9 months respectively. CONCLUSION: Synchronous carcinosarcoma of the prostate is a highly malignant neoplasm that may need an aggressive therapy. PMID- 10829450 TI - [Diffuse involvement and rapid progression of high-grade transitional carcinoma in a kidney transplant recipient]. AB - In renal transplant patients, an increased risk of cancer has been seen, higher than in the general population, and the more frequent genitourinary neoplasms are the kidney and bladder ones. Moreover, in those patients the pattern of tumor growth is very rapid, with early tendency to local and systemic dissemination. The case here presented, a patient with kidney transplant 3 years before, with high grade, superficial transitional cell carcinoma involving the whole bladder, the pelvis and the ureter of the right native kidney, shown this fast progression. Four months after radical cystectomy plus bilateral nephroureterectomy, the disease progressed to intraperitoneal and hepatic metastases. PMID- 10829451 TI - [Penile cutaneous horn. A clinical case]. AB - Contribution of a case report of cutaneous horn of penis surgically treated with extensive resection of the implantation base. A well differentiated, microinvasive epidermoid carcinoma was histopathologically demonstrated on a hyperkeratosis squamous papilloma. Although underlying lesions to cutaneous horn are usually benign, malignant changes have been reported in up to one third of cases; surgical treatment should therefore include extensive resection of the implantation base. PMID- 10829452 TI - [Transvesical repair of non-complicated vesicovaginal fistula]. AB - OBJECTIVE: The management of vesicovaginal fistula still remains controversial in regard to the timing of repair and type of approach. We review the relevant literature and describe our experience with the transvesical management of uncomplicated vesicovaginal fistulas after gynecological surgery. MATERIAL AND METHODS: We reviewed retrospectively 6 consecutive women who presented with uncomplicated vesicovaginal fistula, repaired at our institution between 1995 and 1998. Four cases underwent early repair after a trial of conservative management. One case underwent intraoperative repair and another case after 19 months because of delayed diagnosis. Postoperative follow up ranges from 4 to 35 months. RESULTS: All patients were cure. The fistula tract was excised totally in all patients. There were no postoperative complications. CONCLUSIONS: Early repair of uncomplicated vesicovaginal fistulas after gynecological surgery offers reliable success, according with literature records and our own experience. On the other hand, transvesical repair offers an easy operative approach, with a high successful rate. PMID- 10829453 TI - [Cystine calculi. Prevention with captopril. Clinical case]. AB - Contribution of a case report of multiple cystine lithiasis with underlying homozygous cystinuria in a 5-year old male child. Following surgical management of the lithiasic episode the patient was followed-up with super-hydration, urine alkalinization and captopril prophylaxis. After three and a half years evolution, the patient has not developed new lithiasic episodes and maintains normal dibasic amino acids values in 24 h urine. PMID- 10829454 TI - [Abdominal-pelvic actinomycosis with urinary tract involvement, secondary to gynecologic infection caused by intrauterine device]. AB - Abdomino-pelvic actinomycosis is a condition caused by Actinomyces israelii, a Gram-positive opportunistic bacteria that triggers and develops the infection only in previously injured tissues, and then slowly progresses and spreads until it extrinsically affects the urinary tract. Use of an intrauterine device is a known risk factor to suffer from this disease. Relative risk in IUD users is two- to four-fold higher compared to IUD non-users. Risk increased with prolonged IUD use. Treatment is by removal of the causative agent, surgical resection of necrotic tissues and administration of intravenous Penicillin G, 4 million units every 4 hours for 30 days, followed by Amoxicillin 500 mg every 8 hours for 12 months. This paper contributes two cases of abdomino-pelvic actinomycosis with urinary tract involvement in IUD users. Standard treatment was employed with good evolution. PMID- 10829455 TI - [Silica urolithiasis: report of a new case]. AB - We report a new case of silica urolithiasis in a 71-year-old patient with background of medicament ingesta (antacid) which could justify the formation of these calculi with so uncommon composition. Medicament calculi amounts about 1% from total urinary calculi. Among them, silica ones (silicon dioxide or Si O2) have been relate din their etiology to chronical ingest of antacid drugs, specially those with magnesium trisilicate. Silica formation in urine is facilitated and conditioned by an acid pH of it. Diagnosis and therapeutical approach does not change in relation to other lithiasis with similar dimension or location. After its composition report, and ruling out the possibility for patient's malingering, urine alkalinization was applied, with an excellent follow up afterwards. The antacid was substituted by other drug. We analyze the most important aspects about this case. We also make reference to the main related bibliography. PMID- 10829456 TI - [The value of troponins in emergencies]. PMID- 10829457 TI - [Myocardial necrosis is not observed during acute cocaine abuse detectable by the serum levels of troponin T]. AB - BACKGROUND: Recently we can use a new family of serological markers to objective myocardial damage with hig sensibility and specificity. One of these markers is T troponin (c-TNT). Some cocaine abusers have chest pain during drug consumption due to myocardial ischemia produced by the drug. OBJECTIVE: To study if in the chest pain appeared after cocaine consumption a c-TNT increased is objective, showing myocardial cellular damage. PATIENTS AND METHODS: We studied 65 patients; 35 cocaine abusers that were admitted into the emergency room for chest pain after cocaine consumption and 30 patients with no coronary chest pain. To all of them were determined c-TNT and CK. RESULTS: An increase in c-TNT or CK was not observed in both groups. CONCLUSION: We don't observe an increase in c-TNT in the group of cocaine abusers that evidence myocardial cellular damage. PMID- 10829458 TI - [The prevalence of antithyroid antibodies in Lleida]. AB - OBJECTIVE: To evaluate the prevalence of autoimmune thyroid antibody, goiter and levels of ioduria in the region of Lleida. METHODS: Cross-sectional study on a randomized sample of 601 individuals above 6 years old. Examination and palpation of thyroid gland, TSH, T4, FT4, T3, peroxidase antibodies, thyroglobulin antibodies, and ioduria. RESULTS: The prevalence of positive peroxidase antibodies was 3.4% (IC 95%: 0.9-5.9), and that of positive thyroglobulin antibodies 9.7% (IC 95%: 6.5-12.9). Palpable goiter was found in 18.3% of individuals, and visible goiter in 13.3%. The mean ioduria was 120.2 +/- 68 mg/l. The mean TSH value was higher in the individuals with thyroid autoimmunity (3.4 +/- 2.4 mU/l) than in the rest (1.6 +/- 1.2 mU/l) the difference was significant. CONCLUSIONS: Although cases of hypothyroidism are common in populations with iodine deficiency, in regions with adequate iodine intake justifies monitoring for hypothyroidism in these populations as well. PMID- 10829459 TI - [Giant-cell arteritis: a descriptive study in southwestern Spain]. AB - OBJECTIVE: To study the clinical and laboratory features of a series of patients with giant cell arteritis (GCA) or temporal arteritis in south-western Spain (Extremadura). PATIENTS AND METHODS: Retrospective study of 25 patients with GCA diagnosed by temporal artery biopsy between 1990 and 1998. RESULTS: Nine patients were males and 16 (64%) females. Sixteen cases (64%) presented polymyalgia rheumatica (PMR). Other clinical findings were: fever/febricula (64%), constitutional syndrome (64%), new headache (96%), visual symptoms (48%), jaw claudication (17%) and abnormal temporal arteries (17%). All patients had an ESR of more than 50 mm/hour and a raised C-reactive protein. Thirteen patients (52%) had anemia (hemoglobin level < 12 g/dl). Eleven cases (44%) presented a platelet count higher than 400,000/mm3. Four patients (16%) had an elevated AST and/or ALT levels and 8 patients (32%) had an elevated GGT and/or alkaline phosphatase levels. In patients with PMR, there was a higher frequency of constitutional syndrome (81 vs 33%, p = 0.02). In females, there was a higher frequency of anemia (75 vs 11%, p < 0.01), platelet count higher than 400,000/mm3 (75 vs 0%, p < 0.01) and elevated AST and/or ALT (25 vs 0%, p < 0.01) and elevated GGT and/or alkaline phosphatase (50 vs 0%, p < 0.01) levels. CONCLUSIONS: The clinical and laboratory features of GCA in our series of patients in south-western Spain are similar to that described in other spanish populations, with the exception of a slightly higher frequency of PMR and a lower frequency of jaw claudication and abnormal temporal arteries. In our study, the clinical picture of GCA was more severe in patients with PMR and in females. PMID- 10829460 TI - [Hospital readmission for heart failure in a general hospital]. AB - OBJECTIVE: Heart failure (HF) is the most common discharge diagnosis for admission and readmission in adult. Prior studies have reported that elderly patients who survive hospitalization for HF, are particularly vulnerable to readmission. The present study was realized to investigate the characteristics of patients with readmissions for HF and the features of this hospitalizations. METHODS: This observational study, looked for patients with two or more hospital admissions with diagnosis of HF in 1996 in Gregorio Maranon General Hospital (Madrid). From them, it was taken a random sample. Second and following admissions were studied (readmission). RESULTS: There were 1561 hospitalizations for HF in 1996, being 349 second or following admissions (22.4%). After analyzing 153, it was found a 75.8% of readmissions in the first 90 days after discharge. The mean age was 77.4 +/- 10.31. More than a half were women and patients have an important comorbidity (pathologies presented more frequently were Diabetes Mellitus and airways obstructive diseases). From patients with echocardiography performed before readmission, 66.4% present left ventricular systolic dysfunction and 33.6% have normal left ventricular systolic function. The principal aetiologies were coronary artery disease, valvular heart disease and hypertension (in those patients in which aetiology have been investigated). CONCLUSION: Readmission after a hospitalization for HF is common, with a high percentage of patients readmitted in a short time. This rate demands efforts to identify patients with a higher likelihood of readmission and develop programs to clarify the determinants of this problem and thus improve it. PMID- 10829461 TI - [Adverse reactions to antihypertensive drugs]. AB - OBJECTIVE: In chronic therapies, the less number of adverse effects (AE) to drugs is decisive to improve the rate of performance and reach the protective objective. We can make comparisons in the rate of tolerance of antihypertensive drugs (AD) if we study all of them with the same method in patients with real hypertension. METHODS: Analyzed 922 hospitalized hypertensive patients looking for adverse effects (AE) reported systematically and spontaneously form, the causality must to excel in two algorithms the uncertain grade. RESULTS: 13.47% of antihypertensive drugs presented an AE, beta-blockers presented more AE than diuretics and angiotensin-converting enzyme (ACE) inhibitors. CONCLUSIONS: The AD haven't the same AE neither feature nor frequency. If to dependent on a poor fulfillment, we lose the protective effectiveness, the application independently of the economic cost isn't right. PMID- 10829462 TI - [The value of the polymerase chain reaction in cerebrospinal fluid for the diagnosis of herpetic encephalitis: a report of 2 cases and a review of the literature]. AB - Brain biopsy has been considered the gold-standard test for the diagnosis of herpetic encephalitis. However, because of its cruence, other non invasive methods have been developed for its diagnosis. In recent years polymerase chain reaction (PCR) in cerebrospinal fluid has been employed for the diagnosis of herpetic encephalitis (HE). We report two cases of clinically suspected HE in which DNA of herpes simplex virus was amplified by PCR from cerebrospinal fluid, and review the role of this method for the diagnosis of HE. Two patients were admitted to the emergency room presenting clinical pictures and cerebrospinal fluid samples suggestive of lymphocytic meningoencephalitis. Both were admitted in the intensive care unit because of altered level of consciousness, beginning empiric treatment with intravenous acyclovir as HE was suspected. Clinical outcomes were favourable in both cases, being confirmed the presence of herpes simplex DNA in cerebrospinal fluid by PCR in the two patients. We have reviewed in the literature the value of PCR for the diagnosis of HE in comparison with other ancillary tests as brain biopsy and serology. PCR could reach according with some authors a sensibility of 98% and a specificity of 94%. PMID- 10829463 TI - [Parathyroid cysts. Their differential diagnosis from thyroid pathology. A report of 2 cases]. AB - Parathyroid cysts are rare in clinical practice, but, when they appear, they can be mistaken for more common thyroid conditions, that can incidentally be associated with them. We present two cases of parathyroid cysts. One of them was misdiagnosed of thyroid nodule and the other was found in the context of a normo functioning multinodular goitre. In this last case, the presenting symptoms had raised the suspicion of malignancy. Both of them responded favorably to evacuation of the liquid by means of a puncture-aspiration with a fine needle (PAFN). Our clinical cases show that the diagnosis of parathyroid cyst should be considered in patients with a cervical mass, even if an evident thyroid condition is present. They may be accurately diagnosed by means of PAFN and determining PTH in the cystic liquid, whose characteristics help to predict its side of origin. PMID- 10829464 TI - [The clinical response to interferon-gamma in a patient with chronic granulomatous disease and brain abscesses due to Aspergillus fumigatus]. AB - Chronic granulomatous disease (CGD) is an heterogeneous inherited disorder where the microbicidal phagocytic properties of leukocytes are impaired. Patients with CGD are prone to severe ious with many microorganisms such as Aspergillus spp (1,2). Recently, interferon gamma (IFN-8) which significantly reduces the frequency of infections in these patients, has been proved useful (3,4). We present our experience with the clinical use of these cytoscine. A 26-year-old woman with CGD suffered recement multiple brain abscesses by Aspergillus fumigatus treated in many occasions with surgical drainage. Since treatment with IFN-8 was started, two years ago, she is well and asymptomatic until the moment. PMID- 10829465 TI - [Nodular pulmonary sarcoidosis, a radiological form of rare presentation or granulomatous vasculitis? A report of 3 cases]. AB - Three cases of nodular pulmonary sarcoidosis are presented. Chest roentgenograms demonstrated multiple bilateral nodules and the diagnosis was achieved by means of biopsy specimens after other diseases capable of producing a similar histologic or clinical picture were excluded we discuss the clinical, histologic and radiographic behaviors of nodular sarcoidosis. PMID- 10829466 TI - [From the laboratory to the clinic: CFTR and the therapeutic options for cystic fibrosis]. AB - Cystic fibrosis is a common lethal heritable disorder, caused by a defect in a chloride channel protein, namely CFTR. After the identification of the gene and its product by positional cloning (on chromosome 7), CFTR has been characterized as a low conductance (8-10 pSiemens) linear chloride channel, which appears to regulate other apical transport proteins. Two therapeutic options are reviewed: gene transfection and drug therapy. So far, clinical studies have shown that gene transfection cannot effectively restore CFTR function. Simultaneously, several drugs including genistein, phenylimidazothiazoles and gentamicin have been found to activate mutant CFTR, thus, being suitable for single or combined (with gene transfection) treatment. PMID- 10829467 TI - [Levothyroxine treatment of the solitary thyroid nodule]. AB - The aim of this study was to review all published randomized clinical trials evaluating the efficacy of suppressive therapy with levothyroxine for solitary thyroid nodules. No significant differences were observed in reducing the volume of benign solitary cold thyroid nodules between the placebo and levothyroxine groups. PMID- 10829468 TI - [The plasma concentration of factor VIII as the principle risk indicator for venous thromboembolic disease and ischemic stroke]. PMID- 10829469 TI - [Still's disease in a 66-year-old man]. PMID- 10829470 TI - [Menstrual arthritis?]. PMID- 10829471 TI - [Stevens-Johnson syndrome secondary to carbamazepine mediated by photosensitivity]. PMID- 10829472 TI - [Fever with exanthema and meningism: a form of presentation of acute human immunodeficiency virus infection]. PMID- 10829473 TI - [Chylous ascites as a form of presentation of gastric cancer]. PMID- 10829474 TI - [Recurrent pericarditis and colchicine]. PMID- 10829475 TI - [Thoughts on a supposedly paraneoplastic cerebellar syndrome]. PMID- 10829476 TI - [Acute colchicine poisoning]. PMID- 10829477 TI - [Bacterial oligo-polyarthritis]. PMID- 10829478 TI - [Pleural empyema due to Gemella morbillorum with a favorable outcome]. PMID- 10829479 TI - [A new case of intravascular infection by Salmonella enteritidis]. PMID- 10829480 TI - [Vagal chemodectoma with atypical cells. Case report]. AB - We report a clinical case of vagal chemodectoma with cell atypies, which was presented as slowly growing neck mass without any other symptoms. At the beginning and due to the histologic report of the needle biopsy, which morphologic study based on cellular atypia was misinterpreted as a malignant tumor and with the images obtained by computed tomography (CT) suggesting a malignant neoplasia, we proceeded to approach the case as a metastatic tumor of unknown origin. We made a revision of the epidemiologic factors, and diagnostic and therapeutic management of this type of growths. PMID- 10829481 TI - [Microcytic carcinoma of the nose cavity. A case report]. AB - Tumors of the nasal cavity and sinuses are rather infrequent, being squamous carcinoma the predominant type, which accounts for the 80 percent of the totality of them. Maxillary sinus is the most affected, followed by nose cavity. Half of cases are originated on turbinates. We present the clinical case of a woman patient diagnosed of microcytic carcinoma of the right nasal cavity originated at the level of middle turbinate. The mass filled the width of the fossa getting in touch with the nasal septum, without encroaching it. According its extent the tumor was classified as T2, N0, M0 and treated with radiotherapy only. Microcytic carcinoma is an exceptional histological type in nasal cavity and next to its numeric meagerness in this location makes this case a very peculiar one. PMID- 10829482 TI - [Thyroglossal cyst associated with squamous cell carcinoma of the larynx]. AB - Cyst of the thyroglossal duct results from a developmental anomaly of the thyroid gland. There are many congenital neck masses only outnumbered by benign lymphadenopathies. The majority appear in children between 2 und 10 year-old, but nearly a third of cases does not manifest clinically until much later (20 year or so). In association to a carcinoma is an uncommon event, but there are published malignant cases. The AA. report the case of a 74-year-old patient with a neck cystic mass evolving during a 40-year-term associated to a laryngeal squamous carcinoma. The imagery shows the radiological and anatomopathological correlation of changes in the tissues interpreted as a cyst of thyroglossal duct and a squamous carcinoma of the larynx, independent of each other. PMID- 10829483 TI - [Disability and vestibular pathology]. AB - OBJECTIVE: To probe the ability in differentiate the handicap degree among patients with peripheral vestibular illness by means of a questionnaire carefully thought with this purpose. DESIGN: Prospective study covering 2 years (May 1st. 1991 till May the 1st. 1996). FIELD OF STUDY: Patients from the Health Area of Zamora, ambulatory attended at Otolaryngology Department, complaining of peripheral vestibular troubles. INDIVIDUALS INCLUDED: 60 patients diagnosed as paroxitic positional vertigo or Meniere's disease, lasting the discomfort either 6 weeks or less or 2 years or more, the totality split into 4 groups, of 15 individuals each. SCORING: Analysis of the performed scores of the first valuation by means of the Questionnaire for Assessment of the Disability in Vestibular Pathology (Q.A.D.V.P.). Descriptive statistics were performed for each group and each scale. RESULTS: Patients with more than 2 years of evolvement present themselves, usually, with higher scores in both emotional and functional scales if compared with those resulting from patients suffering from paroxistic positional vertigo. CONCLUSIONS: The Q.A.D.V.P. is a measure instrument very useful, by allowing the differentiation of several degrees of disabled people of our studied samples. PMID- 10829485 TI - [Laryngeal leiomyoma]. AB - A case of a 41-year-old man with a simple leiomyoma is reported. The pathological features of these tumors are described and also the therapeutic ones. A review of the literature confirmed that the laryngeal leiomyoma is a very infrequent neoplasm and only 36 cases have been reported previously. PMID- 10829486 TI - [Maxillary sinusitis caused by Aspergillus]. AB - Naso-sinusal aspergillosis is an infrequent ailment which true incidence is not correctly assumed and furthermore the cases are increasing because of the progression of immunodeficiency problems. Suspicious become accentuated when repeated antibiotic treatments fail or in patients with previous dental pathology or radiologically show inside of sinus images of forcing bodies. Despite of which the diagnosis should be histological thanks to the examination of the surgical piece gained following the classic approaches through the canine fossa or endoscopic surgery. The only real treatment is surgery. We report one clinical case of an acute naso-sinusal aspergillosis with ineffectiveness of all used treatments and whose diagnose was achieved only through the anatomopathological study of the piece supplied by the Caldwell-Luc procedure. PMID- 10829484 TI - [Horizontal supraglottic laryngectomy. Study on 41 operated cases between 1989 1996 in La Rioja]. AB - The AA. conduct a retrospective study done with 41 patients operated upon and followed up at the Otolaryngological Department of San Millan y San Pedro Hospital complex, Logrono, between 1989 and 1996. Horizontal supraglottic laryngectomy was the performed surgery, and patients classified in stages I, II, III or IV. The fundamentals of this procedure the oncologic and functional results as well and the survival rate are analysed. Four points are stressed: 1st. The descent of the base of the tongue eases the postoperative swallow. 2nd. The survival shows that recurrences like lymph nodes metastases are the best prognostic factor, with an OR of 2.61. 3rd. More advanced stages have less survival tendency. 4th. The excellence of this technique is evident, having had only one case of local recurrence in our series resolved through totalization of the laryngectomy. PMID- 10829487 TI - [Maxillary mucocele: two case reports and review of literature]. AB - Mucoceles are benign expansive conditions, chronically evolving, from paranasal sinuses. Most frequent localized in the ethmofrontal area and related with sinusal ostial drainage. Maxillary mucoceles are rarer and related with traumatic, tumoral or surgical pathologies, particularly with the Caldwell-Luc incision. These mucoceles demand an aggressive treatment als those etmofrontals in order to achieve a radical exeresis through an outer incision. We report 2 cases of maxillary mucocele. The first was a big one of idiopathic type, the other was associated to an inverted papilloma sitting in the external wall of the affected nasal cavity. Review of the literature in regard of pathogenesis, radiological diagnosis and management of these cases. PMID- 10829488 TI - [Tongue tuberculosis as the first expression of the lung process]. AB - Nowadays tuberculosis incidence seems to increase again after past decades fall. This event has been worsen because of the growth of resistances, basically due to patient's failure to comply with scheduled specific therapeutics. Anyway the decrease of tuberculosis leads initially to lowering of extrapulmonary forms, being the mouth cavity one of the most infrequent seat, so that location can be omitted at first glance diagnosis. We report one tongue tuberculosis case, which was the first expression of a lung tuberculosis, and discuss other manifestations within oral cavity and also the problems of some cases, as well as the best treatment. PMID- 10829489 TI - [Neuroendocrine tumor of the larynx. Small cell carcinoma]. AB - A 69-year-old patient diagnosed, in 1985, of squamous cell carcinoma of the right vocal cord (T1, N0, M0) was treated with cordectomy and radiotherapy. Eleven years later he presented with a larynx tumor of the glotto-subglottic region. Histopathological examination stated a small cells neuroendocrine tumor. We contribute with one new case to those rare laryngeal tumors, which we see often published in the specialty journals, probably due to the wide employed immunohistochemistry dyes. PMID- 10829490 TI - [Fronto-ethmoidal mucocele: endoscopic fronto-orbital approach]. AB - OBJECTIVE: To revise the references of surgical approach of the mucocele into frontal sinus. DESIGN: A retrospective study. STUDY SUBJECTS: Three patients with mucocele frontal and orbitary encroachment were treated with endoscopic and associated external approach. The follow-up period has been between 22 and 36 months. RESULTS: Combined approach allow marsupialization and adequate repermeabilization of the nasofrontal duct. CONCLUSIONS: Endoscopy surgery is the choice method in treatment of mucoceles, but the access has to be completed by an external approach when the lateral wall of the frontal sinus is extended or the mucocele exterioriced. PMID- 10829491 TI - [Pilomatrix carcinoma of the face. Two case reports]. AB - Pilomatrixial carcinoma is the malignant variety of pilomatrixoma. It is a malignant lesion, locally aggressive that can reappear, specially if not completely removed. Scanty are the number of cases listed in the literature (2). Although metastases are uncommon one or twice references can be found in the writings (1). We report 2 cases of pilomatrixial carcinoma located in the middle canthal and malar zones, which were treated surgically: excision of the tumor and reconstruction by means of a local rotation flap. PMID- 10829492 TI - [Pott's puffy tumor]. AB - Between the complications of frontal sinusitis orbital or intracranial are the most frequent encountered (meningitis, abscesses and empyemas). All are secondary to thrombophlebitis of veins communicating the intracranial cavity with the frontal sinus. Frontal osteomyelitis secondary to sinusitis, the so-called Pott's puffy tumor, is a much more rare aftermath in the antibiotic epoch. Pott's puffy tumor must be suspected in patients with frontal headache followed by frontal oedema. Concerning the diagnosis clinical suspicion is essential and must be settled throughout computerized tomography and/or magnetic resonance or even bone scintiscan. The paper report 2 cases, one an orbital periostitis, at the beginning of the disease, which was recovered with medical antibiotic treatment and another one, an osteomyelitis somewhat evolved requiring surgery through frontal osteoplasty. Perusal of etiology, pathogenesis, diagnosis and treatment of this complication. PMID- 10829493 TI - [Comments on the possibility of the use of the ND-YAG laser in chronic hypertrophic rhinopathy]. AB - Impediment of breath through the nose because of turbinal hypertrophy is an often repeated symptom in chronic rhinitis. In the course of years several procedures for clear the nasal congestion, with more or less satisfactory outcomes, has been tested. The introduction of Nd:Yag laser has allowed a new treatment of the hypertrophic turbinates, thanks to the advantage of its conduction through the flexible optical fibers of the traditional optical devices, even connected to the video system. Treatment can be achieved in the outpatients department with local anesthesia. Results are favorable and forebode new horizons in the surgery of chronic hypertrophic rhinopathies. PMID- 10829494 TI - [Goldenhar syndrome: a polymalformation syndrome with conductive hearing loss]. AB - We report the case of a 9-years-old girl showing at examination facial asymmetry, right ear with II degree dysplasy and degree III in the left ear (according Marx classification), cleft palate, renal agenesy, scoliosis and hypoplasy of the left thumb. Otoscopically both external ear canals looked very narrow and very upright positioned. The drumhead image recalling that of chronic mucoid effusion. The measurement of hearing through pure-tone audiometry, PEATC, and tympanometry resulted in a pattern of bilateral transmissive pantonal hearing impairment accounting for 50-60 dB. Scoliosis was due either to bars or hemivertebrae or even to an atlas occipitalization. Chromosoma caryotype was normal. PMID- 10829495 TI - [Esthesioneuroblastoma: report of 2 case]. AB - The esthesioneuroblastoma is a very uncommon malignant growth arising from olfactory epithelium. Almost 400 cases have been published since its was first described. Owing to the meager figures of these series is not possible to drawn out conclusions in order to establish an unique protocol of treatment for this tumoral type. We present 2 cases diagnosed in our Department, each diversely treated and comment the results. PMID- 10829496 TI - [Acute epiglottitis in the adult. Retrospective study of 14 cases]. AB - Acute epiglottitis instead of been a well recognised disease in children, is a rare condition in adult patients. Failure to diagnose epiglottitis early, in adults, undoubtedly contributes to its continuing mortality. We report our experience with acute epiglottitis in grown-up people with a retrospective series of 14 patients seen in our Department from 1993 to 1997. We comment about clinical features and therapeutic management of these patients. PMID- 10829497 TI - [Bilateral aneurysm of the internal carotid artery and thrombosis of the jugular vein. Case report]. AB - Extracranial carotid artery aneurysms is a very odd pathology which first sign usually is a neck mass. At present the arteriosclerosis is considered its main source and surgery is promoted as its regular treatment, but each case must be managed according the distinct peculiarities showed. This paper reports the clinic and radiological features of a woman diagnosed of bilateral carotid aneurysm with the addition of a thrombosis check afterwards of the left jugular vein. Because of the rarity of this case think justified the record. A bibliographical perusal of this singular entity accompanies the article. PMID- 10829498 TI - [Future prospects of laparoscopic ultrasonography]. PMID- 10829499 TI - [Local treatment of rectal cancer]. AB - Transanal local excision is only indicated in small rectal carcinomas with limited infiltration and high differentiation, and is followed by a 5-year survival rate around 90%. Endo-cavity irradiation has similar indications and results, but can only be performed by trained teams. The main risk of these local treatments is tumoral recurrence. A rigorous selection of patients is necessary and a rigorous postoperative survey. PMID- 10829500 TI - [Treatment of adenocarcinoma of the lower esophagus and cardia: resection with or without thoracotomy?]. AB - STUDY AIM: In the treatment of adenocarcinoma of the cardia and lower oesophagus, the choice of the approach (with or without thoracotomy) to perform a proximal oesogastrectomy (POG) is still debated. The aim of this retrospective study was to compare mortality, morbidity and long-term survival in a series of patients operated on with or without thoracotomy. PATIENTS AND METHOD: From January 1991 to June 1997, 59 patients (mean ages: 65 +/- 10 years, range: 30-83) underwent POG through a transthoracic (n = 31) or a transhiatal approach (n = 28). All patients underwent both coeliac and left gastric lymphadenectomy. A mediastinal subaortic lymphadenectomy was only performed in patients who had a transthoracic approach. Both groups were comparable concerning age, weight and height, and tumoral staging according to preoperative imaging and pathologic examination. The transhiatal group included more high-risk patients (respiratory insufficiency, ASA score = 3) (NS). RESULTS: Resection was palliative in four patients in the transthoracic group and two patients in the transhiatal group. Operative mortality was 9% in the transthoracic group and 0% in the transhiatal group (NS). Pulmonary complications were as frequent with and without thoracotomy (35% versus 32% respectively). Global (curative and palliative resections) 3-year actuarial survival was similar in both groups (transthoracic: 39% versus transhiatal: 46%, NS), as well as survival after curative resection (44% versus 49% respectively, NS). The operative approach did not influence survival in patients N+ (22% versus 17% respectively, NS) and in patients N- (86% versus 77% respectively, NS). CONCLUSION: These results suggest that, for adenocarcinoma of the cardia and lower oesophagus, the theoretical carcinologic benefit of mediastinal lymphadenectomy can be balanced with an higher operative risk related to the transthoracic approach. PMID- 10829502 TI - [Treatment of diaphragmatic ruptures by laparoscopic approach in the lateral position]. AB - STUDY AIM: The aim of this retrospective study was to assess the feasibility of the treatment of recent diaphragmatic ruptures through laparoscopic approach in lateral position in patients with stable hemodynamic condition. PATIENTS AND METHOD: From 1994 to 1999, seven patients, 16 to 68 years old, were operated on for diaphragmatic rupture after a 6 hours to 15 days delay. Diaphragmatic rupture was located on the left side in 6 patients, on the right side in 1. Diagnosis was confirmed by CT-scan with reconstructions; there was no associated visceral injury. Patients were operated on by laparoscopic approach in lateral position, herniated viscera reintegrated by moderate pulling and diaphragmatic rupture repaired with non resorbable continuous suture. RESULTS: Stomach was the usual herniated viscera (n = 6); hemothorax was less than 300 cL; diaphragmatic rupture was 8 to 12 cm long. Duration of surgery was no more than 120 minutes. The seven patients after a two months to two years delay had normal chest X-rays. CONCLUSION: The laparoscopic approach in lateral position provides good visibility of the diaphragmatic lesions, easy reduction of herniated organs, complete thorax exploration and cleaning, and easy diaphragmatic repair. This technique is only feasible in patients with stable hemodynamic conditions and does not provide a complete abdominal exploration. PMID- 10829501 TI - [Elective laparoscopic colectomy for sigmoid diverticulitis. Prospective study of 56 cases]. AB - OBJECTIVE: The aim of this prospective study was to assess the feasibility and postoperative advantages of the laparoscopic-assisted elective colectomy for diverticular disease. PATIENTS AND METHODS: From january 1989 to december 1997, among the 114 patients electively operated on for diverticulitis, 56 patients were treated by laparoscopic approach. Evaluated parameters included: gender, age, weight, size, ASA score, operating time, duration of hospital stay, of analgesic treatment, and of postoperative ileus, morbidity and mortality rate. RESULTS: The study group consisted of 35 women and 21 men. Mean age was 59 years (34-81 years); 29 patients were ASA 1 and 27 ASA 2. Overall postoperative mortality rate was 0% and morbidity rate 16% (n = 9). There were no complications directly related to laparoscopic technique. The conversion rate was 14% (n = 8). Mean operating time was 300 min (200-600 min). Mean duration of postoperative ileus was 2.4 days. Mean duration of hospital stay was 9.4 days. CONCLUSION: This study demonstrates the feasibility of elective laparoscopic-assisted colonic resection for diverticular disease in more than 80% of cases with a postoperative morbidity and mortality rate comparable to those of conventional surgery. PMID- 10829503 TI - [Perforated duodenal ulcer: laparoscopic treatment of perforation and ulcerous disease]. AB - STUDY AIM: The aim of this retrospective study was to report a continuous series of 44 perforated duodenal peptic ulcers operated on through laparoscopic approach with curative treatment of the peptic ulcer disease for socioeconomic purpose. PATIENTS AND METHOD: From February 1995 to May 1996, 44 patients were operated on laparoscopically. There were 42 men and two women (mean age: 36 years). All patients had peritonitis with pneumoperitoneum in 68%. Duodenal peptic ulcer was known in 12 patients and antecedent of episodic epigastric pain were present in 27. Four trocads were used. The diagnosis was confirmed by abdominal exploration and peritoneal lavage was performed with physiological serum. RESULTS: The procedures were: suture of perforated ulcer associated with posterior vagotomy and anterior seromyotomy (n = 6), with troncular vagotomy and pyloroplasty (n = 24) and single suture (n = 1). A conversion into laparotomy was necessary in 13 patients (29.5%). There was no mediastinitis, no postoperative death. Peritonitis by leakage occurred in two patients who were reoperated by laparotomy; mean duration of hospital stay was 5.5 days. With a one-year follow-up, all patients were in good condition, free of pain. CONCLUSION: With laparoscopic surgery, diagnosis of peptic ulcer perforation was confirmed, peritoneal lavage was perfectly done, duodenal perforation was sutured and surgical treatment of the peptic ulcer disease was performed, which is important in poor countries. PMID- 10829504 TI - [Supernumerary ectopic hyperfunctioning parathyroid gland: a potential pitfall in surgery for sporadic primary hyperthyroidism]. AB - STUDY AIM: The aim of this retrospective study was to report a series of nine patients with a sporadic primary hyperparathyroidism, operated on for an ectopic supernumerary hyperfunctioning parathyroid gland. PATIENTS AND METHOD: From 1973 to 1998, among a total of 1,307 patients operated on for a primary hyperparathyroidism, 9 (0.69%) had an ectopic supernumerary hyperfunctioning gland. There were six women and three men (mean age: 63 years) with a sporadic hyperparathyroidism. Initial cervicotomy was performed in our institution in 6 cases. The nine patients underwent 19 operations including one through sternotomy. The ectopic parathyroid gland was localized in the eight patients who had preoperative localization studies. RESULTS: The supernumerary gland was located in the anterior mediastinum (n = 6), in the carotid sheath (n = 2) and within the vagus nerve (n = 1). In three patients, it was found during the initial cervicotomy. In the 6 other patients, it was found in the course of a reoperation. With a mean follow-up of five years, all the patients were biochemically cured. One patient had a permanent recurrent nerve palsy and a definitive hypoparathyroidism. CONCLUSIONS: The low incidence of an ectopic supernumerary hyperfunctioning parathyroid gland in sporadic hyperparathyroidism does not justify the routine use of preoperative localization studies and intra operative quick parathormon assay. During an initial conventional cervicotomy the search for a 5th gland is highly recommended when 4 normal glands have been found in the neck. This research should also be performed in case of multi-glandular disease. PMID- 10829505 TI - [Sentinel node and operable breast cancer: utilization of blue dye injection. Pilot study]. AB - STUDY AIM: Sentinel node detection in breast cancer can be realized with colorimetric and isotopic procedures often associated. The aim of this study was to report results obtained with blue dye injection only. PATIENTS AND METHOD: From September 1998 to July 1999, blue dye injection was performed in 73 consecutive patients (mean age: 51 years, range: 36-71 years); 51/70 70% were post-menopausal and half of them were under substitute hormonal treatment; 70% of cancers were discovered through routine mammography. There were 12 bilateral cancers, six of them synchronous, and 84% of cancers were located in the external quadrants. Individualization of sentinel node was performed through blue dye injection into the tumor in case of preoperative diagnosis or in the tumoral site in case of discovery of the cancer through extemporaneous histological examination. RESULTS: 71 out of 73 cancers were classified pT1 and 70% measured 10 mm and over. Individualization of sentinel node failed in two obese patients. Sentinel node invasion concerned one node (n = 7), two nodes (n = 1) and three nodes (n = 1). Conservative treatment was performed in 72 patients out of 73; in case of sentinel node invasion, axillary irradiation was performed without reoperation. CONCLUSION: Blue dye injection for sentinel node individualization is an accurate technique in selected patients in case of small tumors. Reoperation can be avoided and replaced by axillary irradiation in case of N+ tumors. Duration of hospitalization was 48 hours or under in 70/73 patients. Nevertheless isotopic procedure must be recommended as a routine technique in learning centers and for most surgical teams. PMID- 10829506 TI - [Variations in intraperitoneal temperature during laparoscopic cholecystectomy]. AB - STUDY AIM: The aim of this study was to measure thermal variation during laparoscopy in the vicinity of heat sources such as monopolar (MC), bipolar (BC) and ultrasound coagulation (USC) and to evaluate their possible negative consequences for the patient. METHODS: This study included 67 patients who underwent laparoscopic cholecystectomy. The temperature measurements were taken with a sterile thermal probe introduced through a 5 mm trocar, coupled with a recording monitor reading variations between 20 degrees and 80 degrees C. The variation in temperature was measured as a function of the power applied to the electrodes (20 or 30 W) and in relation to the distance (1, 2, 3, 4, and 5 cm) from the electrodes. RESULTS: The temperature varied by 3 degrees for BC, 29 degrees for MC and only 0.2 degree for USC when the distance increased for 1 to 5 cm. Depending on the power delivered, (20 or 30 W or 1 to 5 for USC), the variations were 1 degree for BC, 17 degrees for MC and there was still no variation for USC. CONCLUSION: The use of bipolar coagulation and ultrasonic coagulation associated with minimal temperature variations is the option of choice for operating near structures such as the common bile duct or the gastrointestinal tract. PMID- 10829507 TI - [Morbid obesity: laparoscopic gastroplasty prior to plastic surgery]. AB - OBJECTIVES: Reduction of the weight in obese patients is a pre-condition for the aesthetic improvement of obesity or its after effects by plastic surgery. The aim of this study was to evaluate the effectiveness and advantages of gastric banding, using a ring positioned under laparoscopic guidance. PATIENTS AND METHODS: Thirty-nine patients with morbid obesity were operated on. For the first nine patients, gastric restriction was obtained by Mason's traditional technique. In the remaining thirty, a Kuzmak ring was positioned a few centimeters under the cardia, under laparoscopic guidance. The study concerned the evolution of weight, associated pathologies, and complications. RESULTS: Three months after surgery, all patients had lost weight (mean loss: 15 kg). At six months, a mean weight loss of 26 kg was observed. At one year, the mean weight loss was 35 kg. Two patients experienced postprandial vomiting that justified widening of the ring; one ring was poorly tolerated and was removed 10 days after surgery. CONCLUSION: This new surgical technique is effective for reducing the weight of obese patients. The laparoscopic approach to the stomach reduces parietal complications and enables subsequent abdominal plastic surgery to be performed. PMID- 10829508 TI - [Recurrence of Demons-Meigs' syndrome. A case report]. AB - Meigs' syndrome includes ascites, hydrothorax and benign ovarian tumor. The aim of this study was to report a recurrence of Meigs' syndrome in a 71 year old woman, which occurred 30 years after the initial operation. It seems to be the first reported case of recurrence. The high level of CA125 was suggestive of malignancy. Resection of a peritoneal recurrence of the benign fibrothecoma was followed for the second time by improvement of all the symptoms. PMID- 10829509 TI - [Bilateral post-traumatic adrenal hemorrhage. Report of a case with acute adrenal insufficiency]. AB - Bilateral adrenal haemorrhage of traumatic origin is rarely observed or possibly missed in severely multi-traumatised patients. It can lead to a potentially fatal adrenal shock. Its emergency diagnosis is made by imaging techniques, usually by CT-scan. Early substitution therapy has to be done. This complication emphasizes the importance of an immediate abdominal morphological exploration in multi traumatized patients when this is feasible. PMID- 10829510 TI - [Is ambulatory surgery suitable for cancer patients?]. AB - The aim of this inquiry was to question 50 patients before a traditional hospitalization in a regional center, in order to know their opinion about one day surgery. Among 44 answers from 41 women and 3 men, 29 were against, 11 favourable and 4 without opinion. After their hospitalization, only 4 changed their mind. The main reasons, in case of unfavourable answer, were lack of security and fear of pain. Psychologic aid in traditional hospitalization is frequently cited (34%). This has to be taken into account in ambulatory cases. PMID- 10829511 TI - [C. Walton Lillehei and the epic of open heart surgery]. AB - C. Walton Lillehei (1918-1999) from the Surgery Department of the University of Minnesota in Minneapolis (MN, USA), was the precursor of open heart surgery. He successively introduced heart-lung machine, intracardiac repairs, pacemaker, and mechanical cardiac valves. PMID- 10829512 TI - [Acute primary intestinal intussusception in an adult diagnosed by ultrasonography]. PMID- 10829513 TI - [Voluminous tuberculous adenopathy of the hepatic peduncle]. PMID- 10829514 TI - [Report of a rare complication of evertizing sutures by viscerosynthesis]. PMID- 10829515 TI - [Pellagra-like erythema following vertical banded gastroplasty for morbid obesity]. PMID- 10829516 TI - [Case report of severe primary perineal gangrene]. PMID- 10829517 TI - Chronic pancreatitis: pathogenesis and molecular aspects. AB - Chronic pancreatitis (CP) is characterized by irreversible morphological and functional alterations of the pancreas presenting clinically with upper abdominal pain as well as exocrine and endocrine insufficiencies. CP is morphologically characterized by pancreatic head enlargement, calcifications of the parenchyma, cysts, and pancreatic stones. The most common etiological factor of CP in Western industrialized countries is alcohol abuse; less common factors include hereditary pancreatitis, CP due to metabolic disturbances, CP due to pancreas divisum or duodenal wall cysts, and idiopathic CP. The molecular alterations leading to the chronic inflammatory process are nor completely understood. Research during the last years, however, has elucidated that a number of growth factors and their receptors are overexpressed in CP, which is thought to contribute to the high degree of pancreatic fibrosis and to the proliferative potential of ductular cells in this disorder. In addition, gene mutations have been detected in a subgroup of CP samples underscoring the pre-malignant potential of CP. In this review we will summarize our current knowledge about pathogenic and molecular aspects of CP. PMID- 10829518 TI - Mechanisms of pain in chronic pancreatitis. AB - Pain is a leading symptom in chronic pancreatitis (CP) and often its management necessitates surgical intervention. Nevertheless the presence of different hypotheses, the pathophysiology of pain is not understood, thus the indications for therapy remain controversial. Increased pressure within the ductal system and/or the parenchyma has been suggested to be one of the causes of pain. This controversial theory has been substantiated by the demonstration of a relationship between intrapancreatic pressure and intensity of pain. On the other hand, recent studies have shown the inflammatory involvement of intrapancreatic nerve fibres in a so called "neuroimmune interaction". In fact, infiltration of inflammatory cells around the nerves together with an increase in the number of nerve fibres in the fibrotic pancreatic tissue have been proposed as a possible cause of pain in chronic pancreatitis. Moreover, immunohistological studies have shown that the amount of neurotransmitters, such as substance P and calcitonin gene related peptide, is increased in afferent pancreatic nerves and a close interrelationship between pain and immune cell infiltration of the nerves has been reported in CP. In addition to these hypothesis, extrapancreatic causes such as common bile duct obstruction and duodenal stenosis are discussed. This article review points to the different pathogenic mechanisms of pancreatic pain in CP. PMID- 10829519 TI - Chronic pancreatitis: relation to acute pancreatitis and pancreatic cancer. AB - The relationship between chronic pancreatitis (CP) and other pancreatic diseases, such as acute pancreatitis (AP) and pancreatic cancer (PK), remains a fairly debated question. The progression from alcoholic AP to CP is controversial, and some long-term epidemiological studies suggest that alcoholic CP might be the result of recurrent alcoholic AP (necrosis-fibrosis sequence) and a subgroup of alcoholics may present recurrent AP without progression to CP. Other predisposing factors (genetic, nutritional, environmental) seems to be important in inducing different outcomes of pancreatic damage due to alcohol. However, recurrent episodes of AP are clearly involved in pathophysiology of CP in patients with hereditary pancreatitis. A relationship between CP and subsequent PK development has long been suspected, but we actually don't know whether this association is direct or is the result of confounding factors, such as alcohol intake or cigarette smoking. Many issues should be considered as indicators of a causal association, and several of them are not fulfilled. Nonetheless, epidemiological studies (case-control or cohort studies) showed that the risk of PK is increased in patients with CP; the risk is significantly higher in tropical calcifying CP and hereditary pancreatitis. Studies on growth factors, oncogenes, tumor suppressor genes, and angiogenesis suggest that the sequence PC-KP is plausible from the biological standpoint. PMID- 10829520 TI - Chronic pancreatitis: diagnosis and staging. AB - Chronic pancreatitis is a dynamic disease characterized on one side by a progressive destruction of the pancreatic parenchyma and change in the architecture of the gland and on the other by the impairment of its function. Diagnosis of chronic pancreatitis may be a quite easy or a very difficult attempt according to the severity and evolutive stage of disease. In fact, while most patients presents with a typical history of alcohol abuse, recurrent abdominal pain and steatorrhea, in the late stage of disease it is not rare to see patients with symptoms and signs which may be not typical for pancreatitis. A large number of morphological and functional methods has been developed to allow an easy and early diagnosis of disease. However, while in the advanced stages of disease, where pancreatic insufficiency, calcifications, or pseudocysts are present, diagnosis is easy and most of the procedures show high sensitivity and specificity, in the early disease the degree of pancreatic dysfunction and structural change are too small to be detected by current methods. The present article aims to evaluate the different morphological and functional methods with their advantages and shortcomings, as well as to establish their role in the diagnostic assessment of chronic pancreatitis. PMID- 10829521 TI - Medical treatment of chronic pancreatitis. AB - The medical treatment has an important role in patients with chronic pancreatitis. Pain is the most frequent symptom, at least in the initial phases of the disease. In about 60% of patients it can be successfully treated by medical therapy; in the remaining 40% it requires surgery. Malabsorption of fat and protein and diabetes usually appear in the advanced stages of the disease. The treatment of these complications is based on the administration of pancreatic extracts and insulin. There are several types of pancreatic extracts; the most useful are those with high lipase content and high lipase-protease ratio. Moreover, they should be protected against gastric acid and should have a gastric emptying simultaneously with chyme, with a rapid liberation of enzymes into the duodenum. The treatment of diabetes usually requires low-moderate doses of insulin. Diabetic ketoacidosis is rare, while microvascular changes have the same frequency as in type 1 diabetes. PMID- 10829522 TI - Is there still an indication for pancreatic duct drainage in chronic pancreatitis? AB - Pain in chronic pancreatitis is supposed to be multifactorial in origin. Pancreatic duct/tissue hypertension is today proved in patients with chronic pancreatitis and pain. Duct drainage reportedly normalizes pancreatic duct/tissue pressure and reduces pain in 70% of the patients. Also, duct drainage by endoscopy may relieve pain. Surgical duct decompression is parenchyma-preserving and even suggested to prevent further progress of exocrine insufficiency. Recent experience indicates that such operations are pain-relieving not only in patients with dilated ducts but also in those with small duct disease. PMID- 10829523 TI - Pancreatic pseudocysts in chronic pancreatitis. Surgical or interventional drainage? AB - Pseudocyst formation is a well-known complication of acute and chronic pancreatitis. Many pseudocysts are asymptomatic and may resolve without intervention. For a symptomatic pseudocyst drainage is indicated. Although surgical cystoenterostomy has been the treatment of choice for many years, recently invasive but non-operative treatment methods have challenged surgical drainage as the standard therapy for pancreatic pseudocysts. Both the method as well as the timing of intervention has become a matter of debate. Percutaneous catheter drainage and endoscopic drainage have proven beneficial in the treatment of pseudocysts, although long-term outcome remains to be awaited. Resolution rates after surgical and non-surgical methods are comparable, but clinical and technical aspects may mandate either method. Each patient requires an individual, multidisciplinary approach, thereby obtaining optimal treatment-outcome. PMID- 10829524 TI - Resection in chronic pancreatitis: anastomosis with the jejunum or with the stomach? AB - Even in centers where the first choice in the surgical treatment of chronic pancreatitis is a derivative procedure some selected patients require resection. The most popular solution of gastrointestinal reconstruction still seems to be pancreaticojejunostomy but, the review of the reported experiences, suggests a general trend towards anastomosis with the stomach as a recent policy. A reliable comparison between pancreaticogastrostomy and pancreaticojejunostomy is difficult because the reported series are seldom related to chronic pancreatitis patients only, but are reporting mixed date concerning mainly periampullary cancer. Moreover with only one exception no prospective randomised clinical trails are available; unfortunately the positive trend in favour of pancreaticogastrostomy reported in uncontrolled studies is not confirmed in the randomized setting. Also the comparison between the experiences achieved by the present authors working in centers with different approach to the pancreatic anastomosis does not show statistical significant difference for both morbidity and mortality. In conclusion nowadays the best confidence and experience with any of the two methods represents the basis of choice. PMID- 10829525 TI - Pylorus-preserving Kausch-Whipple resection: the successor of the classical Kausch-Whipple in chronic pancreatitis. AB - Chronic pancreatitis can be a debilitating disease. Resection for complicated disease and in those patients with intractable pain is a major component of therapy. There are potentially important physiological advantages with the pylorus preserving Kausch-Whipple (P-KW) compared to the standard Kausch-Whipple (KW) resection but further prospective studies are required to show this unequivocally. The Beger's operation also represents a significant theoretical advance. Indeed it permits resection in a proportion in whom a PP-KW or KW would not be possible technically because of severe fibrosis around the intra pancreatic portal venous system. Although we have adopted the Beger's operation on our unit, large comparative studies are required against the KW and PP-KW procedures to properly establish its value. The hidden malignancy is an important issue, which undermines the routine use of the Beger's operation. PMID- 10829526 TI - Organ-preserving surgery in chronic pancreatitis: the duodenum-preserving pancreatic head resection. AB - Chronic pancreatitis is a inhomogeneous disease of multifactorial genesis and a variable clinical course. Upper abdominal pain is the leading clinical symptom of the majority of the patients. The primary treatment of these patients is conservative, but if the treatment fails in pain relief or organ complications occur surgical treatment is indicated. The most common organ complications due to chronic pancreatitis are stenosis of the common bile duct and the pancreatic duct, duodenal stenosis, stenosis of the portal vein with portal hypertension, pancreatic pseudocysts and the development of pancreatic fistula. Due to the pathophysiological concept of an elevated duct pressure as a source of pain, duct decompression by drainage procedures is the favored surgical procedure by many surgeons. Nevertheless, even in patients with a dilated pancreatic main duct, only half of the patients will benefit from drainage operations. Long-term severe upper abdominal pain and complications of the neighboring organs due to an inflammatory mass in the head of the pancreas should be indicative for resective procedures which should be organ-preserving as much as possible and take into account the endocrine function of the pancreatic gland. Simultaneous multiple organ resections like pylorus-preserving partial duodenopancreatectomy or total pancreatectomy are not necessary for a benign disease and should be only performed in patients with proven malignancy. The aim of the surgical procedure is to reduce pain and frequency of relapsing pancreatitis without impairing the endocrine function of the pancreatic gland. PMID- 10829527 TI - Modifications of the duodenum-preserving pancreatic head resection. AB - Surgery for chronic pancreatitis has gained wide acceptance because of excellent results regarding pain alleviation and control of complications arising from adjacent organs. After the introduction of the duodenum preserving pancreatic head resection by Beger almost three decades ago, many modifications have been proposed, evaluated and compared. This article reviews the variety of operations, the reported results and potential advantages. Besides the Beger- and Frey procedure, none of the modifications have been properly evaluated in a prospective randomised trial. Both procedures managed to relief the outlined problems while achieving low operative mortality and morbidity. Only the operations according to Beger and Frey can be considered standard procedures in chronic Pancreatitis. PMID- 10829528 TI - What should be the standard operation in chronic pancreatitis: Whipple or duodenum-preserving pancreatic head resection? AB - Surgical options in the treatment of chronic pancreatitis have undergone both development and controversial discussion in the past decades. Operations such as the classical and pylorus-preserving Whipple resections are more and more being replaced by operations such as the duodenum-preserving pancreatic head resection, which preserves extrapancreatic organs like the stomach, the duodenum and the extrapancreatic bile duct. The latter operation preserves a normal food passage and glucose metabolism after surgical intervention. In addition, the duodenum preserving pancreatic head resection provides long-term pain relief and reduction in up to 90% of chronic pancreatitis patients, as well as a general improvement in quality of life. This article will summarize and compare the surgical options in the treatment of chronic pancreatitis and will provide arguments why the duodenum-preserving pancreatic head resection should replace the classical and the pylorus-preserving Whipple resections as the standard surgical procedure used to treat chronic pancreatitis-related complications. PMID- 10829529 TI - [Reflections on mesenteric infarct]. AB - The authors relate their statistic about mesenteric infarction in the last five years and they get out the starting point in order to check the characteristics of a disease, which is today distinguished by an infaust prognosis. After few mentions about etiopathology and pathologic anatomy they keep their attention on clinic manifestations of disease and on diagnostic research. After dealing with medical and surgical therapy they declare how more refined diagnostic technique could help early diagnosis and consequently decrease morbidity and morbidity, which are completely unacceptable. PMID- 10829530 TI - Perforated jejunal diverticulitis: personal experience and diagnostic with therapeutical considerations. AB - A case of perforated jejunal diverticulitis in a 87-year-old man is described and the literature is reviewed. Jejunal diverticulosis (JD) is estimated to occur in 0.02% to 1.3% of the adult population and is found most often in the elderly. The acute diverticulitis with perforation has been reported as high as 2.3% among patients with JD and is associated with high mortality. Clinical presentation mimic other more common acute intraperitoneal inflammatory conditions. Enteroclysis and abdominal CT are the most specific diagnostic tests. The common treatment is surgical resection of the involved segment. Laparoscopic resection and medical and medical/radiological approaches have also been proposed. Diagnostic and therapeutical aspects of this pathology are discussed. PMID- 10829531 TI - Development in diagnosis and treatment of hepatic echinococcosis in a surgical department of a Mediterranean centre over a 20-years period. AB - Liver echinococcosis is an endemic disease in some areas of the world like Middle East and is a serious problem both from social-economic and clinical therapeutical stand-points in other areas like Mediterranean regions. We report our experience on hepatic echinococcosis diagnosis and treatment over a 20-year period. In this retrospective study we have reviewed 89 patients affected by hepatic hydatidosis who underwent surgery in our Unit (between November 1975 and October 1995). Patients were divided into two groups: group A including 65 patients (30 males and 35 females) operated on between 1975 and 1988 and group B including 24 patients (11 males and 13 females) operated on between 1989 and 1995. Main outcome measures reported were recurrence of echinococcosis, association with surgical procedure, complications, diagnostic trial. In group A we performed 41 marsupializations, 20 subtotal pericystectomies and 4 total pericystectomies; while 14 total pericystectomies and 10 liver resections were performed in group B. Postoperative hospital stay was of 30 days for group A patients and of 18 days for group B. A single death occurred in group A. Recurrences occurred in group A only (11 cases equal to 17%) and were treated with total pericystectomies (8 cases) and liver resections (3 cases). Our study shows the development of diagnostic procedures trial during the 20-year period and the different surgical approaches between the two groups. Nevertheless we believe that surgical procedures should be tailored to each patient avoid H high surgical risk due to the benign nature of the disease. PMID- 10829532 TI - [An occult pancreatic insulinoma: radio-guided surgery with 111-In-octreotide. A case report and review of the literature on pre- and intraoperative diagnosis]. AB - INTRODUCTION: Pancreatic insulinoma is a small, rare neoplasm; its radical therapy is surgical enucleation or resection. Although clinical diagnosis is simple, instrumental and radiological localization is still difficult (occult insulinoma: 10-20%). MATERIAL AND METHODS: The authors present the case of a 71 year-old patient suffering from relapsing vertigo syndrome, derangement and transient lypothymia after strong physical activity. ECG, encephalic and abdominal C.T., eco-color-Doppler of epiaortic vessels failed to identify any pathological features. Laboratory tests only revealed that basal glycemia was always lower than normal. Insulinoma was strongly suspected following the outcome of dosages of insulin and C-peptide in the serum and was confirmed after a supervised fast test (72 h). Once the biochemical diagnosis had been reached an abdominal eco, C.T., a selective and superselective angiography and echo endoscopy were carried out to locate the tumor site but results were negative. Only somatostatin-receptor scintigraphy with 111 In-octreotide detected a catchment area (diameter: 3 cm) to the right of the hypochondrium, anterior to the kidney and caudal to the left hepatic lobe. RESULTS: During laparotomy a hand held gamma detecting probe (C-Trak) was used to detect in situ tumour binding of the radiolabelled octreotide and a neoplasm was identified and enucleated from the pancreatic head (diameter: 7 mm), undistinguishable from the surrounding parenchyma. Postoperative signal checks of the surgical field and of the neoplasm revealed, in the first, the absence of captation and, in the second, maximal captation. Histological findings confirmed the nature of the neoplasm: an insulinoma with a ring pattern. Two years after surgery the patient is in good health, with no signs of relapse. CONCLUSIONS: Radio-guided surgery with labelled octreotide makes it possible to pinpoint small insulinomas, locate occult neoplasm, verify the completeness of surgical excision in the field and in the surgical piece, by comparing the different captation of labelled octreotide and check neoplasm multicentricity and any metastases. PMID- 10829533 TI - [Meigs' syndrome: its clinical picture and treatment]. AB - Meigs' syndrome is a rare clinical entity characterised with ovarian benign tumour, ascites and hydrothorax. Between January '94-September '98 we observed three patients with: ovarian neoformation, light (1 patient), moderate (2 patients) monolateral pleural effusion, moderate (2 patients) and considerable (1 patient) ascites. In all patients the preoperative evaluation (sero-haematologic routine, Ca 125 and other oncologic markers, chest X-ray, abdominal and pelvic ultrasonography, total-body Tc, cytological analysis of pleural and abdominal effusion) was suggestive for malignancy but not confirmed it. So an explorative laparotomy with histological extemporary analysis was performed. The results were: 1 fibrothecomas, 1 fibroma, 1 ovarian inflammation with cystic luteinization areas and fibromatosis uterine. All three patients had a good postoperative course. The symptomatology and the effusions disappeared 7-10 days after operation. The first two cases were diagnosed as classic Meigs' syndrome, the third one, instead, as a pseudo-Meigs's syndrome. In conclusion the A.A., according to literature and their experiences, underline: 1) an ovarian mass with pleural and abdominal effusion not always represents an advanced malignancy; 2) even if elevated Ca 125 value is usually associated to a ovarian malignancy, there are some benign lesions in which we observed elevated level of this marker 3) the removal of the ovarian mass is the only resolutive treatment for these patients. PMID- 10829534 TI - [The extempore intraoperative cytology of peritoneal lavage fluid. A study of the reliability of the method]. AB - Data are presented about a 100 cases prospective study, designed to evaluate sensibility/specificity of intra-operative cytology on peritoneal washing in case of cancer of digestive tract. Data analysis showed a very low sensibility of the test (according with most of literature observations) that does not allow to consider it fully reliable. Authors, thus, suggest a critical use of the test and state doubts about his real value in the intra-operative correct managing of therapeutic options. PMID- 10829535 TI - Management of testicular seminoma. Our experience. AB - BACKGROUND: The authors report their experience in the management of seminomas. MATERIALS: They examine 12 cases of seminoma among a series of 19 patients with testicular germ cell tumours observed at the Second Surgical Department of the Second University of Naples. RESULTS: Their results showed a better prognosis for patients in early stage of the disease who underwent surgery and adjuvant prophylactic radiotherapy; good survival rates for patients in advanced stages of the disease were achieved by the combined use of surgery, adjuvant radiotherapy and chemotherapy. DISCUSSION: The authors discuss risk factors, clinical and diagnostic features of seminomas, relating their prognosis with the combined use of both surgery and adjuvant therapies. They consider total orchiectomy, followed by prophylactic radiotherapy, the treatment of choice, especially in stages IA and IIA. They don't perform the routine retroperitoneal lymphadenectomy, differently from American authors, who always achieve it to stage the disease. CONCLUSIONS: The authors stress the improvement in the prognosis of seminoma, which has actually reached the 98% of five-year survival rate, for stages I and II. PMID- 10829536 TI - [Adenocarcinoma of the duodenojejunal flexure. A report of 2 clinical cases and a review of the literature]. AB - The authors report two cases of adenocarcinoma of the duodenojejunal angle and remark the rarity of this pathology, the difficulty of making diagnosis peculiar to neoplasm of the small intestine and the difficulty of treatment peculiar to tumours of the duodenum. According to the literature the diagnosis was determined through X-ray films, after traditional endoscopy was inadequate. Surgical treatment is radical, with extensive exeresis procedures (unless the mesenteric upper vessels are infiltrated) since the prognosis of this tumours is good. PMID- 10829537 TI - [Video laparoscopic cholecystectomy in a female patient with cholelithiasis in Steinert's myotonic dystrophy]. AB - Steinert's disease (SD) is a rare (3-5/100000) myotonic myopathy responsible for chronic restrictive respiratory insufficiency and dilatative myocardiopathy. The authors report the case of a 52-years-old female patient with SD who underwent laparoscopic cholecystectomy for cholelithiasis. Postoperative course was uneventful and the patient was discharged after 4 days. Laparoscopic surgery was effective and safe in the treatment of this pathology. PMID- 10829538 TI - Cystic dilatation of common bile duct. Case report and review. AB - The cystic dilatation of the common bile duct is a rare disease in Europe and in the USA; even rarer in adults. In fact less than 30% of cases are described in the over 20's. There are some reports in the elderly. When observed in adults the diagnosis is usually due to the onset of symptoms of associated pathologies or to complications such as cholangitis or cancer. Ultrasound, CT and ERCP allow, in the majority of cases, an adequate pre-operative characterization of the lesion. Intra-operative cholangiography verifies the completeness of surgery--always resective--and excludes residual biliary pathologies. The treatment in patients who already underwent conservative surgery is more complex, in fact, in symptomatic patients a second observation and resection is mandatory while in asymptomatic patients a careful and complete follow-up is adequate without underestimating the cancer risk. Septic and inflammatory complications, especially if chronic or recurring, can damage the liver; here resection or liver transplantation can be necessary. The case here reported is a 24-year-old woman with recurring cholangitis for at least 2 years in which US, CT and ERCP showed a Todani's type I cystic dilatation of the CBD. The total excision of the cyst with hepatic-jejunal anastomosis was carried out. A two-year follow up demonstrated no symptoms and normal laboratory findings; cholangioscintigraphy showed a good hepatic-biliary and anastomotic function. PMID- 10829539 TI - [Anomalous implantation of the appendiceal base: a clinical case]. AB - The authors report a case of anomalous implantation of the base of the appendix, observed in a 8-year-old boy, arising from the anterior wall of ascending colon at 15 cm from the ileo-cecal valve. After a brief review of the most frequent congenital anomalies of the appendix and the few cases of anomalous implantation described in international literature, the authors emphasize the importance of an accurate exploration and mobilization of the ascending colon to reduce the risk of diagnostic and therapeutic mistakes, also related to the small incision of McBurney, usually made for the routine laparotomic appendectomy. PMID- 10829540 TI - [Measuring stroke volume of the ascending aorta with an extravascular Doppler ultrasound probe in comparison with aortic thermodilution]. AB - Currently, no reliable minimally invasive method of measuring cardiac output continuously in neonates and children undergoing cardiac surgery is available. An extravascular Doppler probe was used to measure cardiac output in 15 New Zealand White rabbits (average weight 3.5 kg, range 2.5-4.5 kg). The results obtained were compared with cardiac outputs determined using the aortic thermodilution principle. The mean cardiac outputs measured with the extravascular Doppler probe was 0.37 +/- 0.01 l/min as compared with 0.39 +/- 0.01 l/min with aortic thermodilution. Regression analysis revealed a close correlation (r = 0.973) between the two techniques. The extravascular Doppler techniques is an option for continuous and reliable cardiac output measurement in small animals used in surgical experiments (open chest models) and in neonates or children during surgical repair of complicated congenital heart conditions. PMID- 10829541 TI - [3-D localization of cardiac structures in real time]. AB - Radiographic 3-D localisation enables measurements to be made that facilitate the placement of the interventional device during cardiac intervention. To enable the reader to implement the method himself, we describe the computation of 3-D coordinates, acquisition of the imaging and projection data on-line, and the accuracy that can be expected with the method. The 3-D coordinates of a cardiac structure are calculated from the image point coordinates, the projection data and the system constants of a biplane isocentric X-ray unit. Technical imaging errors are corrected a priori. The biplane projection data of a run are acquired on-line and stored in a data base. The image pair of interest is identified automatically from the inscribed run number, and assigned to the projection data from the data base. The target image point is marked on the monitor for 3-D localisation. The accuracy of the method was determined by comparing the calculated and actual cross-sectional points of a centimetre grid imaged in biplane X-ray projections. 3-D localisation took an average of 9.8 +/- 1.2 seconds. Angles and distances were assessed with a standard error of 1.1 degrees and 0.8 mm. The run number is identified correctly in 98.5% of the cases. The mean absolute location error for all points and image pairs was 0.61 +/- 0.32 mm. The accuracy and precision was 0.03 +/- 0.40 mm. Radiographic 3-D localisation can be performed readily and accurately on-line. The results obtained with the method enable interventional decisions to be optimized. PMID- 10829542 TI - [Artifact detection and signal reconstruction in the pupillogram]. AB - Just as the EEG reflects different wake and sleep stages, changes in pupil dynamics reflecting different levels of vigilance are also to be found. The literature contains numerous reports on experimental set-ups for the recording of the pupillogram. Interesting methods of signal processing are to be found in [7] and [5]. Currently, such recordings are being used to check the success of sleep therapy. A problem that still needs solving is the optimal handling of artifacts caused by blinking. The present article proposes a procedure for artifact detection by back-propagation networks, and subsequent reconstruction of the signal by an AR model. Estimation of the signal is first demonstrated by a test signal, and then by a corrupted pupillogram. PMID- 10829543 TI - [EMG monitoring in functional electrostimulation]. AB - When using functional electrical stimulation (FES), correct adjustment of stimulation parameters, and monitoring of the stimulated muscle is mandatory if tissue damage is to be avoided. Although several FES systems are already in regular use, a method for direct muscle monitoring is still lacking. This paper investigates the suitability of the electromyogram (EMG) for such a purpose. In six sheep, the right latissimus dorsi muscle (LDM) and the associated thoracodorsal nerve were exposed. Stimulation was effected via electrodes placed on the nerve. Three electrodes were placed in the LDM for EMG recording, and the tendon was connected to a force transducer for isometric force measurement. Stimulation was applied for one second (burst), followed by a three-second pause. The stimulation current was increased in 0.2 mA steps, starting at 0 mA and ending at 4 mA. Throughout the investigation, the EMG signal was monitored with an oscilloscope. In addition, the EMG signal and the force transducer signal were recorded for subsequent analysis. An analysis of the data of all six sheep revealed an almost linear relationship between muscle force and m-wave amplitude (magnitude of r = 0.95, p < 0.001). M-wave monitoring during EMG recording with three intramuscular electrodes is a reliable method of monitoring FES-induced muscle activity, but the absolute force cannot be measured. PMID- 10829544 TI - [Optical radiation and its effect on the skin]. AB - Optical radiation ranges from short wave ultraviolet via visible light to the far infra-red. Both natural and artificial light sources may cause desired and undesired effects, both on the skin and on the organism as a whole. The nature and extent of such effects depend primarily on the energy of the photons, the absorption capacity of the chromophores, the mean depth of penetration, and on the density and dose of the radiation. The present paper discusses the mechanisms of action and desired and undesired effects in the organism, including therapeutic applications, separately for ultraviolet radiation, visible light and infrared radiation. PMID- 10829545 TI - The Morse taper junction in modular revision hip replacement--a biomechanical and retrieval analysis. AB - All biomaterials used for total joint surgery are subjected to wear mechanisms. Morse taper junctions of modular hip revision implants are predilection sites for both fretting and crevice corrosion, dissociation and breakage of the components. The aim of this study is to quantify wear and study metallurgical changes of Morse taper junctions of in-vitro and in-vivo loaded modular revision stems. Three modular revision stems (MRP-Titan, Peter Brehm GmbH, Germany) were loaded by a servohydraulic testing machine. The loads and conditions used exceeded by far the values required by ISO-standard 7206. The tests were performed with maximum axial loads of 3,500 N to 4,000 N over 10-12 x 10(6) cycles at 2 Hz. Additionally, the female part of the taper junctions were coated with blood and bone debris. The free length of the implant was set to 200 mm. One other MRP stem was investigated after retrieval following 5.5 years of in-vivo use. All contact surfaces of the modular elements were assessed by visual inspection, optical microscopy and scanning electron microscopy (SEM). The degree of plastic deformation of the male part of the morse taper junction was determined by contouroscopy. None of the morse taper junctions broke or failed mechanically. Corrosion and wear affected all tapers, especially at the medial side. The retrieved implant showed no cracks and the amount of debris measured only one third of that for the stems tested in-vitro. The present retrieval and laboratory investigations have proven, that the morse taper junctions of the MRP-titanium stem are stable and resistant to relevant wear mechanisms. The longevity of the junctions for clinical use is given. If an optimal taper design is selected, the advantages of modular femoral components in total hip revision arthroplasty will outweigh the possible risks. PMID- 10829546 TI - [Cleaning validation of the prefermentation-fermentation group for production of biomedical products]. AB - The fermentation-group is largely employed to produce pharmaceuticals by means of biotechnology techniques. The operative functions of these plants are surely kept under control by the validation process, while its cleaning is less investigated. The aim of this work is the cleaning validation of a fermentation-group. Furthermore, Standard Operative Procedures (SOPs) have been validated relating to the production necessities and to the allowed safety limits. PMID- 10829548 TI - Effect of dika fat content of a barrier film coating on the kinetics of drug release from swelling polymeric systems. AB - Reservoir drug release systems were produced by coating drug loaded Carbopol 934P disks with polymer films containing Eudragit RL and dika fat at varying proportions. Uncoated disks produced maximal drug release after 5 hrs at pH 1.2 or 13 hrs at pH 6.8. Under these conditions, drug release occurred mainly by gel erosion and/or Fickian diffusion. Coating the disks with films of the polymer blends, predictably, led to more controlled drug release rates. But the ratio of the polymers in the coatings was found to be the modifying factor in the drug release mechanism. Systems containing less than 50% dika fat in the film coatings had a predominantly zero order release mechanism. PMID- 10829547 TI - Synthesis and biological activity of new potential antimalarial: 1H-pyrazolo[3,4 b]pyridine derivatives. AB - The appearance of drug resistant Plasmodium falciparum malaria necessitates the search for novel antimalarial agents. Using the classical ring-bioisosterism concept as a strategy to develop new potential drugs, 1H-pyrazolo[3,4-b]pyridine 4-aminomethanol compounds were designed and synthesized as isosteres of the classical quinoline antimalarial mefloquine. The hydrochloride form of these compounds were tested for in vitro antimalarial activity against chloroquine sensitive (Sierra Leone D-6) and resistant (Indochina W-2) clones of P. falciparum. The results described herein indicated that 1-H-pyrazolo[3,4 b]pyridine system represents a bioisosteric framework to quinoline system in the antimalarial activity. PMID- 10829549 TI - Polylactide microspheres for the modified release of corticosteroids. AB - A steroidal drug, prednisolone 21-acetate, has been incorporated into polylactide (PDLLA) microspheres using a single emulsion/evaporation technique. This paper describes the method used and the characterization of the microspheres obtained: morphology, particle size distribution, drug content, yield of production and in vitro drug release behaviour. PMID- 10829550 TI - Interference of thimerosal in the bicinchoninic acid protein microassay. AB - In this investigation the total amount of protein present in the three influenza vaccine formulations Fluvirin, Fluzone and Flushield have been tested where thimerosal is present as a preservative. Interaction of thimerosal with protein has been confirmed by analysing ovalbumin as a control against ovalbumin thimerosal solution. The results indicate that interference of thimerosal can be overcame for the quantitative assay of proteins by BCA technique with the good reproducibility. The present study identified also the degree of interference of thimerosal in the protein formulations and development of a method to nullify the interference by removing thimerosal. PMID- 10829551 TI - Statistical optimization of transdermal drug delivery system of terbutaline sulfate by factorial analysis. AB - A matrix type of transdermal drug delivery system of terbutaline sulfate was designed and developed by full 3(2) factorial method using polyvinyl alcohol (PVA) of medium molecular weight and polyvinyl pyrrolidone (PVP K-30) as matrix forming polymer and propylene glycol as plasticizer. Patches were designed to deliver 62 micrograms/cm2/hr of the drug into the systemic circulation. These were evaluated for in vitro skin permeation study through excised guinea pig skin. The permeation followed Higuchi kinetics, as its coefficients of correlation (r = 0.965-0.978) predominates over other permeation kinetics. Various physico-chemical parameters of the patches were investigated. Statistical optimization of in vitro permeation rate of the drug from the films were evaluated using two way analysis of variance (ANOVA), regression analysis. PMID- 10829552 TI - Changes in [3H]citalopram binding in cerebral cortex after antidepressant treatment in monoamine-impaired rats. AB - The [3H]citalopram binding after three weeks vehicle, desipramine 10 mg/kg or citalopram 5 mg/kg treatment was studied in the cerebral cortex of normal, DSP-4 , and p-CPA-impaired rats. The DSP-4 50 mg/kg treatment decreased the affinity (Kd), but increased the maximal number of the apparent binding sites (Bmax) of the 5-hydroxytryptamine transporter (5-HTT). This effect was reversed by desipramine 10 mg/kg treatment. The p-CPA 350 mg/kg treatment decreased the Bmax value while the antidepressant treatment did not influence this parameter. In conclusion, our experiments demonstrate that the monoaminergic impairment induced by DSP-4 and p-CPA treatment evokes opposite changes in the 5-HTT binding characteristics and these changes are partially reversed by the chronic antidepressant treatment. PMID- 10829553 TI - Detection of halofantrine, mefloquine and proguanil by charge--transfer complexation on thin layer plates. AB - A complexation phenomenon between certain antimalarials (as donors) and chloranilic acid (as an acceptor) has been studied using thin-layer chromatography (TLC). The antimalarials were subjected to charge--transfer complexation using chloranilic acid as the pi-acceptor. This technique was used for the qualitative detection of the drugs. Drug-spotted TLC plates, when developed in suitable solvent systems and sprayed with the pi-acceptor gave a spontaneous purple colour. The Rr values of the three drugs were found to be constant and specific for each drug sample in a particular solvent system. PMID- 10829554 TI - Visioning together for leadership. AB - The "Vancouver experience" engendered great interest, generating palpable energy and enthusiasm. There were clear messages concerning what we need to know, what we need to learn and how we need to make this awareness a reality. This was a powerful opportunity to share ideas and to build on natural passion and beliefs. Now the task is to ensure that actions are coupled with these insights, resulting in a rich and vibrant future for Canadian occupational therapists. PMID- 10829555 TI - Integration tutorials and seminars: a creative learning approach for occupational therapy curricula. AB - This paper, the first of two companion papers, describes a creative learning approach. Integration Tutorials and Seminars were developed to address concerns of regional fieldwork-education coordinators and preceptors about the ability of third-year student occupational therapists to integrate and apply academic and theoretical knowledge during fieldwork. This ability can be gained through experiential learning in the academic setting and is essential for the effective transfer of academic learning into the students' fieldwork-education experiences (Dale, 1994; Fidler, 1996; McCaugherty, 1991; Neistadt, 1996). The Modified Learning Model of Svinicki and Dixon (1987) was used as a template for an academic course fostering experiential learning. An important secondary goal was to nurture student self-directedness in learning using the philosophy of the Staged Self-Directed Learning Model (Grow, 1991). Case studies were used as a vehicle for engaging and challenging the students. The philosophy, guidelines and process of the Integration Tutorials and Seminars are presented and have the potential to be adapted for occupational therapy curricula around the globe. PMID- 10829556 TI - Integration tutorials and seminars: examining the integration of academic and fieldwork learning by student occupational therapists. AB - This paper, the second of two companion papers, describes a collaborative research study conducted by academic and fieldwork educators of an undergraduate educational programme in occupational therapy. The study examined the perception of student occupational therapists' ability to integrate and apply academic and theoretical knowledge in fieldwork education sites following the introduction of Integration Tutorials and Seminars to the curriculum. The literature review supports the need to develop educational strategies to improve this integration process. The qualitative research design consisted of guided interviews with preceptors and students during two consecutive fieldwork education experiences. Inductive data analysis revealed three themes that contribute to understanding students' ability to integrate academic learning during their fieldwork education: the fieldwork learning environment; preceptor influence on student learning; and the student learning process over time. The discussion section raises implications for academic and fieldwork education approaches. PMID- 10829557 TI - Perceived efficacy and goal setting in young children. AB - One of the central tenets of client-centred occupational therapy is to enable clients to select goals to work on in therapy (Law, 1998). The process of identifying and prioritizing goals is fairly abstract, therefore occupational therapy goals for children are often prescribed by the therapist or by parents and teachers. The purpose of this study was to pilot test a measure and a process that would provide young children with the opportunity to assess their performance on daily tasks and to establish goals for occupational therapy intervention. Parents and children completed the Perceived Efficacy and Goal Setting System (PEGS), a measure of children's perception of their competence performing fine and gross motor tasks. Children 5-9 years of age were able to discriminate among tasks and to rate whether or not they were able to perform each task competently. They were also able to use this information to select and prioritize goals for intervention. While parents often rated the child's competence lower than the child did, there was a high level of agreement regarding which tasks were difficult for the child. Parents and children often did not agree about the specific selection or priority of these tasks for intervention, however, which highlights the need for further research. PMID- 10829558 TI - [Comparison of behavior characteristics of the spontaneously alert state and the liberated state in the term newborn]. AB - Occupational therapists evaluate the capacities of infants at younger ages than ever before. The variability of the infants' state has an inevitable influence on the expression of their behaviours. In order to counter the effect of this instability, Grenier (1985) suggests inducing the "liberated state," which optimizes the infant's level of attention. The aim of this study was to identify the behaviours that discriminate this liberated state from the alert state (state 4) as described by Brazelton (1973). Some of the parameters that were defined in the Synactive Theory of Development Model (Als, 1982) were used in order to document the behaviours of the infants observed in the two different states. The average age of the infants, at the time of observation, was 62 hours. The duration of the observation was 5 minutes. Among the 54 newborns that were evaluated, 21 were observed in the two states under study; 33 were evaluated in one state only. The results confirm a greater stability of the "liberated state". Stress and self-regulation behaviours, and the need for external regulation are less frequent in this state; these differences are statistically significant (p < 0.05). Goal-directed movements are more frequent in the "liberated state" (p < 0.001). The authors suggest the use of the "liberated state" to further document the early capacities of infants. PMID- 10829559 TI - Development of the Handicap Assessment and Resource Tool (HART). AB - An important determinant of whether people can live in community settings is the absence of significant handicap. People with considerable disabilities can live without handicap if they have adequate supports. Handicap, rather than disability, limits peoples' residence options. Disability assessment tools are commonly used to guide where people can live--these assess neither the resources available nor the personal-care handicap present. The Handicap Assessment and Resource Tool (HART) was designed to provide information about the personal-care issues (clothing, hygiene, nutrition, mobility, safety, residence and supports) relevant to choice of residence. The HART was tested by occupational therapists who are frequently expected to provide recommendations regarding disabled clients' residence options. It is a client-centred tool that addresses key occupational performance components of personal care. Pilot testing in hospital and community settings shows the HART is a comprehensive and practical tool that is acceptable to users and clients. PMID- 10829560 TI - A review of a major driver rehabilitation centre: a ten-year client profile. AB - Driver Rehabilitation Services at Bloorview MacMillan Centre in Toronto provide important assessment and rehabilitation services for persons with various physical, neurological, and medical disabilities. Each year, the programme sees approximately 700 to 800 clients. Since 1985, the earliest year for which records are currently available, the program has seen approximately 7300 clients. The client information filing system consists predominantly of paper reports filed alphabetically by hand. A comprehensive computer database program was implemented in 1996 to organize and analyze client-driver records more efficiently. A systematic random sample consisting of 364 records (5% of the total population) was examined to describe the characteristics of the population within five major study categories: demographics, administration, medical information, visual and perceptual abilities and driving record. PMID- 10829561 TI - [Planning of epidemiologic studies using the example of parasitic diseases]. AB - Epidemiology is concerned with the study of diseases and risk factors for diseases in populations. Livestock related studies require that the term disease is extended to cover all conditions that directly or indirectly may affect the economic, social and ecological value of livestock or constitute a public health risk. The objectives of epidemiologic studies range from the quantification of traits, e.g., prevalence of a disease in a population (descriptive approach), to the analysis of correlation between diseases and putative risk factors (analytic approach). The type of epidemiologic study must be selected accordingly. In this paper we present the most important types of epidemiologic studies using example data. Our aim is to give an orientation for the planning of epidemiologic studies. PMID- 10829562 TI - [Ultrasound guided surgical reduction of twin pregnancies in the horse up to the 46th day of pregnancy]. AB - Methods for elimination of one of two embryos in twin pregnancies of mares were studied. Transvaginal ultrasound guided punction of yolk sac or allantoic cavity with fluid aspiration was used. Punction was done in 8 mares, pregnant between 22 and 46 days. In six cases, pregnancy did continue normally with one embryo only while in two cases, both embryos died following the procedure. PMID- 10829563 TI - Efficacy of ELISA for detection of antibodies against several Ornithobacterium rhinotracheale serotypes. AB - The present investigation were carried out to compare self-made ELISA based on SDS-Antigen extraction of serotype B and one commercial ELISA-kit (Biocheck, Gouda, The Netherlands) for their ability to detect antibodies against 12 ORT serotypes (A-L). Using both ELISA systems, antibodies against all serotypes were detected. Examination of serum samples collected from commercial flocks showed similar results on flock bases. However, some minor variations on sample basis could be demonstrated. In conclusion using both ELISA system antibodies against different serotypes could be detected. PMID- 10829564 TI - Dermatophilosis: a threat to livestock production in Ethiopia. AB - A retrospective study of dermatophilosis was made in livestock (cattle, sheep, horses and goats) of Ethiopia under different situations. The disease was reported for the first time in sheep (8.5%), goats (3.14%) and horses (2.84%) from Ethiopia. The occurrence of the disease in cattle was significantly (p < 0.05) higher in wet (6%) than in dry (3.6%) season and in cross (26.9%) than local (3.2%) breeds. The highest prevalence (35%) was noted in cross breeds kept under poor management in the rainy season. The threat of dermatophilosis to animal production in Ethiopia is indicated. Environmental and management risk factors associated with the occurrence of the disease in Ethiopia are identified. The identified risk factors in this study may contribute to the knowledge of devising an optimal intervention in the control of the disease. PMID- 10829565 TI - Pharmacokinetic properties of florfenicol in Egyptian goats. AB - The single-dose disposition kinetics of florfenicol was determined in healthy, non-lactating Egyptian goats, after its intravenous (i.v.) and intramuscular (i.m.) administration at 20 mg kg-1 b.wt. Drug concentrations in serum and urine were determined using microbiological assay method and data was subjected to a kinetic analysis. Florfenicol concentrations in serum decreased in a bi exponential manner after intravenous administration with distribution (t1/2 alpha) and elimination (t1/2 beta) half-lives of 10.256 +/- 0.938 and 56.237 +/- 3.102 minute, respectively. The steady-state volume of distribution (Vdss) and total body clearance (Cltot) were 3.413 +/- 0.304 l kg-1 and 3.306 +/- 0.333 l kg h-1. After intramuscular administration, the peak serum concentration (Cmax) was 0.859 +/- 0.025 micrograms ml-1, achieved at (Tmax) 1.220 + 0.045 h. Florfenicol was detected in urine up to 24 and 96 hour after i.v. and i.m. administration, respectively. The extent of the protein binding and systemic bioavailability of florfenicol were 22.45 +/- 1.727% and 65.718 +/- 3.372%, respectively. PMID- 10829566 TI - Antibacterial efficacy and pharmacokinetic studies of ciprofloxacin on Pasteurella multocida infected rabbits. AB - Twenty four isolates of Pasteurella multocida were taken from 74 diseased rabbits which suffered from respiratory manifestations with an incidence 35.1%. All isolated strains were identified biochemically and serologically as Pasteurella multocida type 3. The in vitro sensitivity test proved that Ciprofloxacin was the most effective antibiotic which was used for treatment of diseased rabbits. There was no isolation of Pasteurella multocida organism from all treated rabbits with Ciprofloxacin which can eliminate the causative organism, and control the infection within the examined rabbit colony. The kinetics of Ciprofloxacin, a fluoroquinolone, was studied after multiple oral and I/V dose of 20 mg/kg b. wt. In healthy rabbits and in those diseased with Pasteurella multocida Ciprofloxacin was given at a dose (20 mg kg-1/day) orally for successive 5 days. Serum levels reached their peak Tmax, at 2.321 and 2.524 hrs. in healthy and infected rabbits after oral administration, with absorption half-life (t0.5 (ab)) of 1.22 and 2.41 hrs. and elimination half-life (t0.5 (B)) of 2.24 and 1.28 hour respectively. Following I/V injection the kinetics of Ciprofloxacin follow two compartment model with (t0.5 (B)) 1.201 and 0.82 hour for normal and infected rabbits, (Vd (area)) of 0.653 and 0.563 L/kg and total body clearance CL(B) of 0.385 and 0.358 L/kg/hr respectively. High tissue concentrations of the drug were recorded in the kidneys, lung, spleen, liver, and muscle of diseased rabbits, while in healthy animal group the highest concentrations were achieved in kidney, liver, lung, spleen, and muscle. The high Ciprofloxacin tissue concentrations indicate that the Ciprofloxacin may be an excellent drug for treating urinary and respiratory tract infections. PMID- 10829567 TI - [Legal report: expert opinion. Can the induction of abortion in heifers mated accidentally and too young be expected from the owner?]. AB - The objective of the presented expert opinion is the reasonability of a termination of unwanted pregnancies for the owner of young heifers mated accidentally by a bull. Because the heifers were too young when conceiving they showed retarded growth and dystocia when delivering after normal course of pregnancy. These problems could have been avoided by pregnancy diagnosis and subsequent induction of abortion in time. PMID- 10829568 TI - Ultrasonographic study on the growth and dimensions of healthy children and adults organs. AB - We measured by ultrasound the spleen, the kidneys, the pancreas in 323 children (age 0-12), the thyroid in 60 children of same age range, and all these organs in 180 adults (in adults we measured additionally the testis, the gallbladder and the choledochus). Children and adults were all healthy, without evidence of pathologies potentially involving these organs. The children have been selected looking at clinical and hematological parameters, while the adults have been selected among a military population, that is the most significant sample of healthy young adults. We measured the length, the transversal diameter and the thickness of the spleen, kidneys, pancreas, testis and thyroid, the diameter of the choledochus and the maximum length of the gallbladder (pre and post stimulus). We found a good correlation between age and dimensions for pancreas, kidneys and spleen in children, representing the progressive growth of these organs. Our data represent an assessment of the normal dimensions of these organs in vivo by means of ultrasound, and therefore they are an useful tool to discriminate pathologically enlarged or reduced organs, both in children and in adults. PMID- 10829569 TI - Neck motion evaluation after whiplash: a radiographic and kinematic protocol. AB - Evaluation of functional damage is the main aspect in the diagnosis of whiplash injury, therefore some type of instrumental testing is strongly needed. We describe an instrumental protocol to test subjects with whiplash, to support clinical evaluation, in order to solve the difficulty of evaluating the functional injury and the lack of fixed parameters capable of identifying this kind of injury. Eighty subjects suffering from after effects of whiplash and seventy five normal subjects undergo two instrumental tests: kinematic examination of neck mobility, and measurements of extension-flexion X-rays of the cervical spine. The results exhibited by the traumatized patients have been compared with the normal range of motion obtained from the data bank of the normal subjects. This is a report in progress of a larger research on whiplash evaluation in a clinical setting. PMID- 10829570 TI - Lack of effect of aerial ammonia on atrophic rhinitis and pneumonia induced by Mycoplasma hyopneumoniae and toxigenic Pasteurella multocida. AB - The objective of this experimental study was to determine the effects of aerial ammonia on disease development and bacterial colonization in weaned pigs inoculated with toxigenic Pasteurella multocida and Mycoplasma hyopneumoniae. Two groups of 10 pigs each were continuously exposed to 50 and 100 p.p.m. ammonia, respectively, and compared to a non-exposed control group of 20 pigs. Following aerosol inoculation with M. hyopneumoniae at day 9, all pigs were aerosol inoculated with toxigenic P. multocida type A at days 28, 42 and 56. At day 63 they were euthanized. Clinical signs including coughing and respiratory distress were present in all groups following inoculation. No significant differences could be established in the extent or frequency of pneumonia between ammonia exposed pigs and controls, or in the extent of conchal atrophy, the frequency of isolation of toxigenic P. multocida from conchae, tonsils, lungs and kidneys, or the average daily weight gain. The recovery of toxigenic P. multocida from nasal swabs following inoculation was significantly greater in pigs exposed to 50 p.p.m. ammonia or more as compared to the control group. In conclusion, high levels of ammonia combined with inoculations with M. hyopneumoniae and toxigenic P. multocida had no significant effect on disease development, but may have enhanced colonization by toxigenic P. multocida on the nasal turbinates. PMID- 10829571 TI - Immunohistochemical detection of antigen in lamb tissues naturally infected with sheeppox virus. AB - The present study describes the detection of sheeppox virus antigen in various lamb tissues, using an immunohistochemical technique, in sheeppox cases which occurred naturally. Sheeppox viral antigen was detected in the cytoplasm of sheeppox cells and degenerated epithelial cells of the skin, lungs and digestive tract involving typical sheeppox lesions. Nuclear staining was also observed in some typically deformed nuclei of sheeppox cells. The immunostaining of sheeppox virus showed a correlation with the presence of sheeppox cells and degenerated epithelial cells resembling them. Additionally, in order to confirm the presence of sheeppox virus in the skin and lung samples, direct electron microscopy was performed and sheeppox virus was only demonstrated in two skin samples. PMID- 10829572 TI - Canine distemper virus causes apoptosis of Vero cells. AB - Apoptosis of Vero cells infected with two canine distemper virus (CDV) vaccine strains was detected using TdT (terminal deoxynucleotidyl transferase)-mediated dUTP nick end-labelling (TUNEL), flow cytometric analysis, agarose gel electrophoresis and electron microscopy (EM). By TUNEL, apoptotic cells were found in CDV-Onderstepoort (CDV-Ond)-infected cells. DNA fragments isolated from infected cells were separated by agarose gel electrophoresis and a 'ladder' pattern appeared. EM observations demonstrated that the cells undergoing cytopathic effect (CPE) possessed morphological characteristics of apoptotic cells. Flow cytometric analysis indicated that CDV could induce apoptosis of Vero cells, but the percentages of the apoptotic cells were correlated with the CPE types. The strain showing the cell-rounding type of CPE produced a much higher percentage of apoptotic cells than CDV-Ond with the syncytium type of CPE (P < 0.01). It was concluded that CDV vaccine strains could induce apoptosis of Vero cells and the apoptosis was virus strain-dependent and cell-dependent. The mechanism remains to be studied. PMID- 10829573 TI - Antibacterial susceptibility patterns of Pseudomonas strains isolated from chronic canine otitis externa. AB - Development of antibiotic resistance in bacteria is a problem of great concern. It is important to establish the convenience of antimicrobial susceptibility tests in animal infections. The aim of this study was to test the susceptibility to antibiotics of Pseudomonas strains isolated from chronic canine otitis externa. We tested 23 strains of Pseudomonas: 19 Ps. aeruginosa, three Ps. fluorescens and one Pseudomonas spp. The most effective antibiotics were tobramycin (100% susceptible), marbofloxacin (91.3%) and ceftazidime (91.3%). Ticarcillin and gentamicin, commonly used for the treatment of otitis externa also showed good results (susceptibility of strains was 86 and 65.2% respectively). Lower susceptibility was found using enrofloxacin (52.1%) probably due to its indiscriminate use. We emphasize the need for a rational policy of antibiotic prescribing in order to prevent the selection of resistant strains. PMID- 10829574 TI - Experimental intravaginal infection of goats with caprine herpesvirus 1. AB - Three adult goats, seronegative to caprine herpesvirus 1 (CpHV.1), were intravaginally inoculated with BA.1 strain of CpHV.1. The animals were kept under observation for 1 month and daily both clinical examinations and white blood cell count were performed. Ocular, nasal, rectal and vaginal swabs and heparinized blood samples were collected every day to attempt virus isolation on cell cultures and detect viral DNA by polymerase chain reaction (PCR) assay. The virus was isolated and detected by PCR only from the vaginal swabs for 5-7 days post infection. The animals showed transient fever and leukopenia and typical necrotic lesions on the vulva and vagina. PMID- 10829575 TI - The detection of proviral DNA by semi-nested polymerase chain reaction and phylogenetic analysis of Czech Maedi-Visna isolates based on gag gene sequences. AB - A semi-nested polymerase chain reaction (snPCR) for detecting proviral DNA of ovine lentivirus (OvLV) in peripheral blood mononuclear cells was developed. Primers for snPCR were situated within the gag gene of the Maedi-Visna virus (MVV) genome. A comparison between the snPCR and serological tests (agar gel immunodiffusion test, immunoblot) were performed using 98 ovine blood samples. Thirty (30.6%) of the 98 sheep examined had antibodies specific for the MVV. PCR showed 21 of them to be positive and nine seropositive animals to be PCR negative. Six of the 68 serologically negative sheep were found to be PCR positive, probably due to delayed seroconversion. The PCR amplification products of these six sheep were sequenced and subjected to phylogenetic analysis. The resulting phylogenetic tree of partial gag gene sequences confirmed that the ovine lentivirus genotype in the Czech Republic is more closely related to the prototype MVV isolates than to the caprine arthritis encephalitis viruses. PMID- 10829576 TI - Prevalence of antibodies to Leptospira serovars in sheep and goats in Alto Adige South Tyrol. AB - Serum samples from 313 sheep and 95 goats were collected during November 1993 in 26 localities in Alto Adige-South Tyrol and tested by microscopic agglutination test for antibodies to 28 serovars of the genus Leptospira. At the time of blood collection all the animals appeared healthy with no clinical sign suggestive of leptospirosis. The observed seroprevalence in sheep was 6.1%, whereas the seropositivity rate for goat serum samples was 2.1%. The highest serological prevalence in sheep was recorded for serovar castellonis, followed by poi, sejroe, hardjo subtype hardjobovis, copenhageni, and cynopteri. Titres to poi were the only ones found in goats. These findings, which are proof of Leptospira infection in Alto Adige-South Tyrol, indicate that foci of several serovars exist in this region. PMID- 10829577 TI - Report of a case of bronchopneumonia associated with Moraxella bovis isolation in a chamois (Rupicapra pyrenaica). AB - A case of fibrinopurulent bronchopneumonia associated with Moraxella bovis infection in a chamois (Rupicapra pyrenaica) is described. The animal, a 4-month old female, was referred by the staff warden of the National Game Reserve of Freser-Setcases (Catalonia, north-eastern Spain). The animal was in good general condition and was found 4 h before death. On necropsy the lungs were congested and oedematous, with haemorrhagic areas in the cranial and middle lobes. The microscopic lesions were those of a fibrinopurulent bronchopneumonia. Microbiological study of the samples obtained showed numerous small beta haemolytic colonies in pure culture, identified as Moraxella (Moraxella) bovis. PMID- 10829578 TI - Evaluation of radiology as a tool to diagnose pulmonic lesions in calves, for example prior to experimental infection studies. AB - The aim of the study was to evaluate radiology as a technique to visualize pulmonary lesions in young calves, e.g. as a selection criterion for research animals in order to eliminate animals with lung lesions prior to experimental studies of pneumonia. Five calves with acute clinical signs of pneumonia were included in a direct comparative study of radiological and post mortem findings. Also, a number of animals with no signs of pneumonia were included as controls. The study revealed good agreement between the radiological and post mortem findings. Thus, in conclusion, radiology should be considered as a useful objective tool to predict the presence of pulmonary lesions in young calves. PMID- 10829579 TI - The Isagoge of Pseudo-Soranus. An analysis of the contents of a medieval introduction to the art of medicine. PMID- 10829580 TI - [History as natural science: Ernst Hallier and Emil du Bois-Reymond as cultural historians]. PMID- 10829581 TI - [Megavolt roentgen site of the Hamburg-Barmbek General Hospital (1938-1945): from therapeutic device to radiation weapon]. PMID- 10829582 TI - Quantitative evaluation by image analysis of platelet morphological modifications after contact with polyvinylacetate. AB - BACKGROUND: In order to investigate platelet activation after contact with artificial materials, which is an important aspect of biocompatibility especially for the blood-contacting devices, platelet morphological modifications and spread area were evaluated by light microscopy and image analysis after contact with glow discharge-treated polybutylene terephthalate coated with a polymer for platelet concentrate filtration. METHODS: A hydrophilic polymer made of partially hydrolyzed polyvinyl-acetate containing polyethylene oxide/poly-propylene oxide copolymer block as lateral chains (PVA) (Biofil S.r.l., Cavezzo, Modena, Italy) was evaluated. After contact with PVA, platelets were allowed to settle on a siliconized slide and then fixed and stained. The specimens were analyzed by image analysis. The percentages of spreading, round and dendritic shapes, as well as the presence of aggregates, were evaluated, and the mean area of the spread platelets was measured. RESULTS: PVA induced significant variations neither in the percentages of shape change distribution, nor of the mean spread area. However it determined a statistically significant reduction in platelets with the area from 60 to 70 mu 2. Such minimal variations agree with the results we obtained in the past, namely a non significant platelet adhesion induced by the same material. CONCLUSIONS: The method confirms the results of platelet adhesion and release reaction (the study of release reaction needs more refined but more expensive methods). However, the study of morphological modifications by image analysis is not suitable for testing materials that induce massive platelet adhesion, because the number of the residual platelets could be too low for the microscopic evaluation. PMID- 10829583 TI - Prospects of prostanoid therapy. Preliminary results. AB - BACKGROUND: To evaluate the efficacy of treatment with prostaglandins (Iloprost and Alprostadil) in patients with peripheral arterial disease in whom a surgical act or a redo operation is not proposable and to set the role of these drugs in the therapy of arterial diseases. METHODS: We analyzed the results of 96 treatments in patients with peripheral arterial diseases of different etiology, treated in the Day Hospital in the Division of Vascular Surgery from the 1/1/1993 to the 30/6/1998. The efficacy of the treatment has been evaluated on the basis of the clinical picture (healing or reduction of ulcers, reduction of rest-pain, improvement of the ability to walk). RESULTS: We had positive results in 60% of the patients who completed the treatment, a good outcome for patients with particularly severe arterial disease. Interruptions induced by the occurrence of uncontrolled side-effects are a few (6.25%). Better results have been found in a special category of disease (arterial diseases not associated with diabetes, not very advanced clinical picture, female gender). Iloprost proved to be superior to Alprostadil. CONCLUSIONS: Prostaglandins today represent an effective therapeutic solution to be considered. Further studies need to be done to find a category of patients who can have the best benefit from this kind of drug. PMID- 10829584 TI - Carotid artery dissection. Correlation of different diagnostic techniques. AB - Dissection of the carotid artery is a rare entity, but a frequent cause of cerebrovascular accidents in young adults and can result in severe neurologic consequences that are often irreversible. Carotid dissection can be secondary to trauma, brisk rotation of hyperextension of the neck, as a complication in certain diseases. It may lead to distal embolization, complete occlusion of the vessel, extensive or limited stenosis, aneurysmal form which has the appearance of a berry aneurysm or resolution of the process with complete patency of the dissected vessel. The authors find color-Doppler sonography to be an invaluable tool in the evaluation of such a process, since it is non invasive, readily available, relatively inexpensive and can yield detailed information about the lumen of the vessel, blood flow, mural and intramural structures. Furthermore, in comparison to MRA, color-Doppler sonography is particularly suitable for the early detection of thrombus. Ultrasonographic investigation has some methodologic limitations: occlusion cannot always be demonstrated directly because of its high cervical location and detection of aneurysms localized in the retrostyloid++ space is usually not possible. The combination of ultrasonographic findings and MRA is the method of choice for follow-up on evaluation of carotid dissection and helps make the decision as to whether one should proceed to angiography in this young age group. PMID- 10829585 TI - Calcium antagonists in the therapy of acute myocardial infarction. AB - The theoretical background and experimental evidence of the use of calcium antagonists in the treatment of acute myocardial infarction are summarised. The main clinical trials that have studied the effects of the three different groups of these drugs in patients with myocardial necrosis are then reviewed. Finally, the conclusion is drawn that there is no proof of the efficacy of calcium blockers in the therapy of patients with acute myocardial infarction but that, in some subgroups of these subjects (Q-wave myocardial infarction with good left ventricular function, non Q-wave myocardial infarction), non-dihydropyridinic compounds should be effective, in particular in patients with contraindications to beta-adrenergic blockade. PMID- 10829586 TI - The damage to a person caused by venous thromboembolism in the civil responsibility. AB - The venous thromboembolism can clinically show itself as deep venous thrombosis or as pulmonary embolism. Both serious and potentially fatal, for this high incidence, they assume importance in social economic sphere. The authors take into account the medicolegal diagnostics methodology of the deep venous thrombosis and of the pulmonary embolism, the traumatic and post traumatic etiology, to determine the connection of causality and the estimating parameters of the damage to a person in the sphere of civil responsibility. To attain to a certain diagnosis of thromboembolism, since its difficult cause of paucisymtomaticity or asymtomaticity of the pathology after an attentive evaluation of symptoms, clinic manifestations and factors of risk, it can't be disregarded to utilize scientific diagnostic criteria, and instrumental ascertainments, serial too, helped by conventional means of standardization, such as the new American system of classification CEAP. The following phases of medicolegal ascertainment consist in identifying the causal connection between disease and event and in estimating of the damage to a person, with rigorous and objective methodology and using tabular orientation guides, that have to indicate the percentage incidence of the undergone disablement on the person's validity for indemnity. It is showed the particular delicacy of the medical examiner's evaluation in thromboembolic disease, in the sphere of civil responsibility, both for the difficulties of the diagnostic identification of the deep venous thrombosis, and of the pulmonary embolism, and for the determination of the connection of causality with traumatic events and with following operation of orthopedics-traumatology and neurosurgery (sector on which the most difficult problems of professional responsibility can connect) and finally for the real evaluation of the consequent damage to a persons, in order to its indemnity. PMID- 10829587 TI - Streptococcus pneumoniae infections: effective prevention soon? PMID- 10829588 TI - Long-term follow-up results of vesico-ureteral reflux treated with subureteral collagen injection (SCIN). AB - BACKGROUND: Endoscopic sub-ureteral injection is at the present a widely used technique for the treatment of vesico-ureteral reflux and it has a good range of efficacy, from 50% up to 90% in relation to different grades. In this paper we report our 1-7 year follow-up results of sub-ureteral collagen injection (SCIN). METHODS: Between 1991 and 1997 we treated with SCIN 129 refluxing ureters in 93 children (60 females and 32 males), mean age 2.1 years (range 3 months-5 years). Fifty-five children had monolateral, but 37 had bilateral reflux. Distribution among different grades was 42 ureters (32.5%) grade 2, 63 (48.8%) grade 3, 24 (18.6%) grade 4. Children were treated with injection of highly purified bovine collagen. In this study all the children underwent radiological follow-up for evaluating the results of treatment with SCIN also in a period of time from 3 to 7 years after endoscopic procedure. RESULTS: After a 1 to 7 years follow-up period we have the following results. In grade 2 we have found absence of reflux in 36 cases (85.7%) after 1 injection and in 38 (90%) after 2 injections. In grade 3, reflux was absent on 46 cases (73%) after 1 injection and in 55 (87.3%) after 2 injections. In grade 4 reflux was cured in 16 (66.6%) after 1 injection and in 18 (75%) after 2 collagen injections, 15% of all units, underwent Cohen's reimplantation. CONCLUSIONS: After 7 years of experience we have concluded that sub-ureteral collagen injection is actually a very good procedure for treatment of mild grade refluxes. We treat these refluxing ureters only when antibiotics prophylaxis is to be prolonged over 6 months with no changes in cystourethrography findings or when recurrent urinary tract infections occur in spite of antibiotics intake. In conclusion, if we evaluate the overall results we see that 73.6% of refluxing ureters have been cured by SCIN, the rate of recurrence is 16.8% and the 15.5% finally underwent Cohen's reimplantation. PMID- 10829589 TI - [The effectiveness and acceptance of a medical device for the treatment of aphthous stomatitis. Clinical observation in pediatric age]. AB - BACKGROUND: To evaluate the efficacy of a new bioadhesive patch, Aloe vera hydrogel, for the treatment of aphthous stomatitis. METHODS: An open, not controlled study was performed in 31 pediatric out-patients, aged 6-14 years, affected by mouth ulcers were enrolled consecutively in the 3 Gps Depts+ of San Marino Republic. For each case, data on case history and clinical profile, patterns of the lesion, presence of spontaneous or provoked pain were collected at baseline, and a bioadhesive patch ("Alovex patch") was administered on the basis of a daily regimen of < or = 3 patches for 4 days. Data on modification of the above-mentioned parameters, with patients and physicians opinion on the therapeutical efficacy, were collected during a control visit (4 days later). Moreover, by means of a daily diary, patients recorded information on the course of the symptoms during the 4 days and were also asked to compare the current treatment with other previous therapies. RESULTS: At the control visit 77% of the patients have shown a marked resolution of spontaneous pain, while in the other patients, pain was significantly decreased to a "mild" or "moderate" level. No one child declared to suffer from severe pain. Also provoked pain resulted to be significantly decreased after treatment Global efficacy was judged positively, being the therapeutical effect in more than 80% of cases "evident or of absolute improvement" both by physicians and patients opinion. A positive improvement of symptomatology started within the 2nd day of treatment in 74% of the patients. The compliance (adhesivity, acceptability and palatability) of the formulation was judged largely favourable in more than 90% of the patients. CONCLUSIONS: The results of this study underline the good efficacy and compliance of the patch for the treatment of the aphtous stomatitis; also the limit of topical available therapies, linked to the "contact time", to develop their therapeutical action, seems not to be evinced on the basis of this study, so the application of this patch seems to be more easy and beneficial. PMID- 10829590 TI - Monitoring the trend of overweight children in Cremona (anni 1990-1998). AB - AIM: To evaluate the trend of infantile obesity in the school-aged population of Cremona over time. METHODS: The weight and height of all children aged 6, 9 and 11 attending schools within the health district of Cremona were measured for 9 consecutive years (January 1990-March 1998), giving a total of 17,730 measurements. Pupils whose bodyweight was 20% above the mean weight for stature and sex given by NCHS canthimetric classes were defined as obese. In addition, three degrees of obesity were defined: slight obesity (20-29% excess bodyweight), moderate obesity (30-50%) and severe obesity (over 50%). RESULTS: This study showed the trend of the increasing prevalence of infantile obesity in the zone in question: it rose from an overall level of 6.1% to 13.6% in 1998. The increase regarded all age groups; levels were three times higher in older children, but obesity had more than doubled in 6-year-olds. With regard to the three classes of obesity, there was a particularly striking increase in moderate obesity, whereas the cases of severe obesity remained constant. CONCLUSIONS: These data underline the need for more incisive educational and preventive work in the field of infantile obesity to be carried out as soon as possible. PMID- 10829591 TI - [Helicobacter pylori infection in pediatrics. Present knowledge and practical problems]. AB - Helicobacter pylori (H. pylori) infection is acquired in childhood, earlier in developing countries, as a consequence the prevalence of infection is higher in developing countries (70%) than in developed countries (5-15%). H. pylori infection spreads from person-to-person, however the precise mode of transmission (oral-oral, fecal-oral or gastro-oral routes) is as yet, not known. Diagnosis of H. pylori infection can be performed with both invasive endoscopic-based tests, or non-invasive tests, mainly by measurement of IgG antibodies against the bacterium in serum samples or by measurement of 13CO2 in expired air (13C-urea breath test). In clinical practice endoscopy and biopsy is recommended before treatment to determine the presence and the degree of gastritis or ulcer. However, endoscopy is a complicated procedure in children and diagnosis of infection can be based on a non-invasive test. The association of H. pylori infection with recurrent abdominal pain seems evident in a subgroup of children with endoscopic features of gastritis, ulcer or hemorrhage. There is an increasing interest in the extraintestinal manifestations of H. pylori infection in children, i.e. iron-deficiency anemia, growth retardation and migraine, but this domain remains controversial. Since infection at a young age is believed to result in chronic atrophic gastritis and gastric cancer in adult life, it is logical to consider a future massive programme of eradication and immunization. Regimens suggested for H. pylori eradication are a combination of inhibitors of gastric acid secretion plus two antibiotics for 7-10 days. PMID- 10829592 TI - Long-term hormone replacement therapy in two patients with Kabuki syndrome and growth hormone deficiency. AB - The Kabuki syndrome is characterized by mental retardation (mild-to-moderate), skeletal anomalies, typical facial appearance and post-natal growth deficiency. The authors describe two patients with Kabuki syndrome and proven growth hormone deficiency. The first patient has been on GH replacement therapy for 4 years; the second for 11 years. On the basis of a sufficiently long follow-up period the Authors discuss the advisability of replacement therapy with growth hormone in patients with Kabuki syndrome. PMID- 10829593 TI - Vein of Galen aneurysmal malformation. Different clinical expressiveness. Three case reports. AB - The vein of Galen aneurysmal malformation (VGAM) is a rare cerebro-vascular disorder in neonates. It is characterized by an abnormal direct communication between one or several cerebral arteries and the vein of Galen. It may appear in the neonatal period or afterwards. Three cases of patients affected by VGAM with different clinical expression are presented. Two of them were treated successfully with endovascular embolization. It was not possible to provide the same treatment to the third patient for an intractable congestive heart failure already existing when VGAM was diagnosed. PMID- 10829594 TI - Asphyxiating thoracic dysplasia in a lethal form: radiological and sonographic findings. AB - The authors report a case of a lethal form of asphyxiating thoracic dysplasia (Jeune syndrome) in a newborn female with a narrow thoracic cage causing severe respiratory failure at birth. The diagnosis was unequivocally confirmed by radiographs of thorax and pelvis. This report emphasizes the importance of the radiological and sonographic features in establishing a correct diagnosis. PMID- 10829595 TI - [Bilateral intravesical ureteroceles associated with hydroureteronephrosis and hyperechogenic spots in kidneys. Report of a neonatal case]. AB - Ureterocele is a cystic dilatation of the terminal ureter. In children it is most commonly ectopic and associated with the duplex collecting system, less frequently it is intravesical and associated with the single collecting system. We present the radiological and ultrasound findings in a male newborn with bilateral intravesical ureterocele associated with hydroureteronephrosis and hyperechogenic spots in both kidneys. In the literature this association has never been reported in neonatal age. PMID- 10829596 TI - Management of autoimmune thyroiditis in childhood. PMID- 10829597 TI - [Heredity, constitution and diathesis]. PMID- 10829598 TI - [The numerus clausus. It is time to inform]. PMID- 10829599 TI - [Tuberculosis associated with the human immunodeficiency virus]. AB - The HIV pandemic is one of the factors that have contributed to the the worldwide increase in tuberculosis cases especially in subsaharian Africa. The copathogenicity between M. tuberculosis and HIV is best illustrated by the high susceptibility of the HIV-infected persons for reactivation of a remote tuberculous infection or early progression of a newly acquired disease and by the negative impact of tuberculosis on natural history of HIV characterised by increased incidence of clinical progression and increased mortality rates. Clinical presentation is rather atypical and severe when immune suppression is advanced: no cavitation on the chest X-rays, visceral ganglionnar involvement with frequent fistulization, positive blood cultures. A standard 6 months antituberculous therapy has the same efficacy as in HIV uninfected patients excepted in severe cases especially if immunosuppression is advanced when 9 months treatment are recommended. Higher frequency of side effects to the antituberculous drugs and drug interactions between the rifamycin and several antiretrovirals due to the induction or inhibition of the CYP3A could interfere with the treatment of the tuberculosis. Paradoxical reaction to the antituberculous therapy after immuno-restoration is induced by the combined antiviral therapy has also been reported. Isoniazid prophylaxis is recommended in patients with a induration of more than 5 mm on PPD skin test and is associated with a 60% reduction of risk of development of tuberculosis. PMID- 10829600 TI - [Post-concussion syndrome: myth or reality?]. AB - The organicity of mild head injury and the postconcussion syndrome are becoming increasingly well documented by neuropsychologic and neuroradiologic studies. The evidence suggests that most plaintiffs are not cure by a verdict. They have cognitive and somatic symptoms, improving with time. Unfortunately for some patients, despite objective recovery, postconcussional complaints persist. Emotional symptoms may have become prominent with time. The real goal in management is to prevent the development of persistent postcommotionnal symptoms by interventions that reduce anxiety, vertigo and headache. PMID- 10829601 TI - [CT-xray demonstration of a vertical vein]. AB - Congenital anomalies of the thoracic veins are infrequent but important developmental disorders. Anomalies of the systemic thoracic veins usually are asymptomatic but may be associated with other more serious cardiovascular abnormalities and complicate their management. We present a case of a vertical vein consisting in an abnormal venous drainage from the left upper pulmonary lobe to the superior vena cava, resulting in a non cyanotic shunt. PMID- 10829602 TI - [Should we give the examination results to the patients?]. AB - Immediately after medical imaging procedures, patients often ask for the results of their examination. Such a request is very uncomfortable for the physician performing the examination, who usually does not know the whole medical file, the psychological status of the patient, how he is informed about his illness, etc. By the means of a questionnaire we asked belgian nuclear medicine physicians about the frequency of such situations and about their practical attitude. 204 questionnaires were sent; we received 119 answers (58%) after a 6 weeks delay. In more than 85% of the cases, this situation occurs regularly, and more than 50% of the participating physicians encounter the problem in more than 20% of the cases. Less than 10% of the participating physicians never give any result to the patient asking for it, while more than 20% always give it. Most of the physicians takes into account the clinical situation of the patient. Unfortunately, the official guidelines do not help to clarify the situation. This work confirms that physicians who perform diagnostic procedures frequently encounter difficulties in patients information. It would be necessary that all people who are concerned by this problem collaborate in order to define a clear and accurate attitude which would be easily applicable in our daily clinical practice. PMID- 10829603 TI - [From university pedagogy to medical pedagogy: some reflections]. AB - Recent studies have shown that an appropriate pedagogic training, characterized by a high specificity, is necessary for teaching in universities. However, the time when it should be initiated and whether it should be a compulsory or a optional formation are still debated. Furthermore, the specificity of medical training is at the origin of a new expending discipline, medical pedagogy, that must take into account the necessity to provide the students with a basic professional competence. Finally, postgraduate continuous medical education has its own methods that also require a preliminary pedagogic training. PMID- 10829605 TI - [Solitary pulmonary nodule]. PMID- 10829604 TI - [Etonorgestrel (Implanon) subcutaneous implant]. AB - Etonorgestrel is provided in an subcutaneous implant to assure a contraception of long duration. This progestogen derived from desogestrel already used in several combined oral contraceptives acts by inhibiting ovulation and modifying the viscosity of the endocervical mucus. The contraceptive effect is rapidly obtained after insertion. After removal of the implant, the serum concentration of etonorgestrel rapidly drops down so that most patients ovulate after 3 weeks. The most important side effect of this contraceptive device is the bleeding irregularity encountered in a minority of patients (frequent or prolonged bleedings). PMID- 10829606 TI - [Bladder hyperactivity]. PMID- 10829607 TI - [Vulvar pathologies]. PMID- 10829608 TI - [Salpingitis and pelvic peritonitis]. PMID- 10829609 TI - [Post-menopausal bacterial cystitis]. PMID- 10829610 TI - [Diagnostic imaging of the head and neck]. PMID- 10829611 TI - [In Process Citation] PMID- 10829612 TI - [Jules Thiriar (1846-1913): the beginnings of abdominal surgery in Brussels]. PMID- 10829613 TI - [Short-term memory performance of German and Chinese probands with verbal and figural items: on the function of the phonological loop and visual-spatial sketchpad]. AB - In a computer-controlled experiment using n = 48 German and n = 48 Chinese subjects we established the memory span for simple geometrical figures and their names in the subjects' respective native language. Half of the subjects replied verbally, the other half nonverbally via a touch screen. In addition, verbal reproduction times were measured. Significant differences in memory span between the two language groups were found only under nonverbal report conditions: here, the Chinese subjects achieved better results than their German contemporaries. There were no differences in oral reproduction times between the two groups. The results are discussed within the framework of Baddeley's working memory model. PMID- 10829614 TI - [Effect of goal setting on information processing in letter comparison tasks]. AB - In two laboratory experiments (N1 = 84, N2 = 20) the impact of goal setting on information processing (perception) in letter-matching tasks was examined. It was expected that setting difficult goals for reaction times improves performance because identification and especially semantic classification of letters should be speeded up under goal-setting instructions. Three findings can be emphasized. (1) Current models of simultaneous letter matching are incomplete because they do not account for differences in reaction time based on alphabetic variations (lower case vs. upper case letters) within a pair of letters that are largely independent of real letter sizes. (2) In both experiments and for all types of letter pairs (AA, Aa, Ab AB), goal-setting effects are found as expected. In addition, the explanation that these effects are due to a speed-accuracy trade off can be excluded. (3) Moreover, in some conditions goal-setting effects are also influenced by special features of letter pairs. Specifically, goal-setting effects are especially strong for letter pairs in which the right letter was enlarged and in which alphabetic differences and semantic differences within pairs call for opposite answers (Aa, AB). Based on the overall pattern of results, it is concluded that difficult goals lead to a general facilitation of processes in visual perception and, in addition, to a reduction of stimulus induced, performance-delaying inhibition processes if linguistic information is processed predominantly in the left hemisphere. PMID- 10829615 TI - [Attitude formation: experienced ambivalence, subjective difficulty and response latency in evaluatively inconsistent, consistent and neutral information]. AB - Ambivalent attitudes simultaneously invoke positive and negative evaluations of a single object (e.g., Thompson, Zanna, & Griffin, 1995). Consequently, ambivalent attitudes towards unfamiliar objects should result from evaluative inconsistency of relevant information about the object (Jonas, Diehl, & Bromer, 1997). The present study shows that the formation of novel attitudes on the basis of evaluatively inconsistent information takes longer than the formation of attitudes based upon positive, negative, or neutral information. This impact of ambivalence should be mediated by the perceived difficulty of attitude formation. Need to Evaluate (Jarvis & Petty, 1996), a chronic tendency to form attitudes spontaneously, is assumed to moderate the level of ambivalence, as well as perceived difficulty and attitude response latencies. The hypothesis was tested in a within-subjects design (N = 30). An unfamiliar object was either described by strongly, moderately, or slightly evaluatively inconsistent information or by positive, negative, or neutral information. Furthermore, links to impression formation are illustrated and differences with regard to attitude formation are discussed. PMID- 10829616 TI - [Frequency and mean amplitude of spontaneous electrodermal fluctuations are not interchangeable indicators of psychological processes]. AB - The study was concerned with the question of whether the frequency and the mean amplitude of spontaneous electrodermal fluctuations convey the same information about mental processes. In order to examine this, the content and the personal relevance of induced imaginations was varied experimentally. The experimental manipulations followed a 2 x 2 factorial design for independent groups (content of cells: 24 subjects each). Changes in electrodermal activity were measured as differences between the imagination and a resting phase. Results revealed that the two variables are not exchangeable indicators of mental processes. Although the two variables correlated significantly, they responded differently to the experimental manipulations. The imagination of death-related contents increased the frequency of spontaneous electrodermal fluctuations more than the imagination of neutral contents. High personal relevance enhanced the mean amplitude of these spontaneous fluctuations regardless of the imagined content. Therefore it was concluded that the two variables have something in common but, on the whole, do not depend on identical factors. Frequency of spontaneous electrodermal fluctuations seems to be a reflection of covert orienting processes, whereas their mean amplitude seems to reflect the activity of an appraisal system incorporating goals and past experiences. However, both variables, seem to be, in the same way, dependent on a kind of basal activation or vigilance. PMID- 10829617 TI - [Phonological information in short-term memory of sentences]. AB - Potter and Lombardi (1990) showed that words that are semantically similar to nouns of auditorily or visually presented sentences intrude into sentence recall if presented in an unrelated word list (given either before or after the sentence). The authors interpreted this finding in terms of the conceptual regeneration hypothesis. This hypothesis states that sentence recall is based on conceptual information whereas the role of phonological information is negligible. However, this general interpretation is questionable. In order to demonstrate the involvement of phonological information in sentence recall, we used Potter and Lombardi's intrusion paradigm. Participants were either presented with auditory or visual material. With respect to visual presentations we used two conditions: a rapid presentation rate (rapid serial visual presentation, RSVP) and a slow presentation rate. Under all conditions the word list was presented before the sentence. The intrusion effect was only found for RSVP but not for the slow conditions. This finding provides evidence for the assumption that phonological information generally plays an important role in short-term sentence recall. PMID- 10829618 TI - Human uses of ultrasound: ancient and modern. AB - For untold millennia certain animals have used ultrasound to probe places where light is unavailable, echo-locating bats being among the most adept. With ultrasonics, bats can quickly and safely 'see' at night in pursuing insects or flying in dark caves. Unable to hear ultrasound, humans have nevertheless made use of it. They did this anciently by taming wolves, with their keen ultrasonic hearing, for aiding in the hunt. Currently, they are doing this by developing technology to detect, generate and process ultrasound for searching in air or other gases, in water or other liquids, and in solids. The story of these technological developments is a large and fascinating mirror of human history involving the advent of such discoveries and inventions as magnetostriction, piezoelectricity, sonar, ultrasonic microscopy, etc.--the list is long. By now we are skilled in probing for underwater objects, the internal structure in materials, organs inside the human body, etc.--again the list is long. A number of different ultrasonic systems can be categorized into one of three key generic approaches: pulse-echo exploration, intensity mapping, and phase-amplitude measurement. In addition, each of these categories can be combined with the others to produce hybrid systems for which an unambiguous categorization is difficult or impossible. Challenging problems remain but solutions are being found. New principles and techniques are being discovered that will improve the use of ultrasound. Employing tomo-holographic techniques to reduce ambiguity in probing three-dimensional objects, near-field techniques to boost resolution and using limited-diffraction beams to provide image construction with ultra high frame rates are cases in point. PMID- 10829619 TI - Research and development of micromechanisms. PMID- 10829620 TI - State-of-the-art surface acoustic wave linear motor and its future applications AB - Two merits of the surface acoustic wave (SAW) device are its high energy density and small size. However, the driving frequency is around 10 MHz or higher. In spite of the difficulties involved with high frequency, the high energy density is attractive for actuator applications. The SAW linear motor's no load speed and maximum output force were 1.1 m/s and 3.5 N using a silicon slider. The silicon slider dimensions were 4 x 4 x 0.3 mm3. We made a lot of 30 microns diameter projections on the silicon surface. The acceleration was 1000 m/s2. The SAW motor is expected to be a high speed, quick response, high resolution microactuator, and much more. High driving voltage was a problem. Our newly designed electrode proved that the driving voltage was reduced to less than 10 V to excite the traveling wave. For actual applications, the SAW device will be placed in a slider. This design is effective in terms of performance and cost. The nanotribology of the SAW motor is also an important and interesting subject. PMID- 10829621 TI - Ultrasonic actuators for nanometre positioning AB - This paper deals with ultrasonic motors based on bimodal plate vibrations and their application to nanometre resolution stepper positioning systems. The concept of a linear ultrasonic motor drive capable of nanometric steps, long range travel and reversible controlled motion is presented. The motor concept developed is based on the superposition of a longitudinal and bending vibrations of a rectangular resonator. The ultrasonic motor model based on system identification via discrete observations and prediction has been developed for control applications. The control algorithm for ultrasonic motors has been developed and theoretical investigations have been made. The open loop positioning system with designed stepper ultrasonic drive produced 10 nm resolution and 5% displacement repeatability. The system with computer controlled position feedback has shown 0.3 micron positioning accuracy over the 100 mm positioning range. PMID- 10829622 TI - Non-contact transportation using near-field acoustic levitation AB - Near-field acoustic levitation, where planar objects 10 kg in weight can levitate stably near the vibrating plate, is successfully applied both to non-contact transportation of objects and to a non-contact ultrasonic motor. Transporting apparatuses and an ultrasonic motor have been fabricated and their characteristics measured. The theory of near-field acoustic levitation both for a piston-like sound source and a flexural vibration source is also briefly described. PMID- 10829623 TI - A cylindrical micro-ultrasonic motor (stator transducer size: 1.4 mm in diameter and 5.0 mm long) AB - Ultrasonic motors are suitable for a micro-actuator because of a direct drive, no brake and their simple construction. In order to fabricate micro-ultrasonic motors, a piezoelectric thin film is essential. In this study, a hydrothermal method was adopted to deposit a PZT thin film. The dimensions of the stator transducer were: outer diameter, 1.4 mm; inner diameter, 1.2 mm; and length, 5.0 mm. The resonance frequency of the stator transducer was 227 kHz, and the vibration amplitude was 58 nmp-p at an input voltage of 4.0 Vp-p. The maximum revolution speed was 680 rpm, and the maximum starting torque was 0.67 microNm. The experimental conditions were 20 Vp-p and 5.3 mN pre-load. The motor property versus scale was investigated using an equivalent circuit. The calculated results indicated that the output torque of the 100 microns diameter motor was 27 nNm. PMID- 10829624 TI - Survey of the present state of the art of piezoelectric linear motors AB - Piezoelectric ultrasonic motors have been investigated for several years and have already found their first practical applications. Their key feature is that they are able to produce a high thrust force related to their volume. Beside rotary drives like the travelling wave motor, linear drives have also been developed, but only a few are presently commercially available. In the present paper, we first describe the state of the art of linear piezoelectric motors. The motors are characterized with respect to their no-load velocity, maximum thrust force, efficiency and other technical properties. In the second part, we present a new motor, which is judged to be capable of surpassing the characteristics of other piezoelectric motors because of its unique design which allows the piezoelectric drive elements to be pre-stressed in the direction of their polarization. The piezoelectric elements convert energy using the longitudinal d33 effect which allows an improved reliability, large vibration amplitudes and excellent piezoelectric coupling. Energy loss by vibration damping is minimized, and the efficiency can be improved significantly. Experimental results show that the motor characteristics can be optimized for a particular task by choosing the appropriate operating parameters such as exciting voltage, exciting frequency and normal force. PMID- 10829625 TI - A matrix model of the axle vibration of a piezoelectric motor AB - In this work, a matrix model of the axle vibration of a piezoelectric motor is proposed. The stator of this motor is composed of a thin piezoelectric membrane and a steel axle fitted at the center of the membrane. The rotor consists of a cylinder-shaped permanent magnet, pressed in contact with the other end of the axle by means of the magnetic forces. A travelling wave is excited in the membrane by using four electrodes and four, properly delayed, driving signals. The rotating flexural displacement of the membrane produces a wide precessional motion of the axle. In this way, a continuous slipping takes place between the axle and the rotor, and therefore, a torque is transmitted to the rotor. In this paper, the precessional motion of the axle is modeled as the composition of two transverse vibrations belonging to two perpendicular planes passing through the axle. The axle, vibrating in its transverse mode, is modeled as a two-port system: the input is the bending moment supplied by the membrane, and the output is the transverse force at the terminal end of the axle. With this model, we have computed the trasmission transfer function as a function of frequency, and the transversal displacement along the axle at its resonance frequency. The computed results are in reasonable agreement with experimental interferometric measurements carried out on a prototype. PMID- 10829626 TI - Temperature self-compensated lithium tantalate piezoelectric gyroscope for higher sensitivity and stability AB - A new H-type gyroscope using a single LiTaO3 crystal that combines the H-type vibrational mode with a tuning fork type vibrational mode is proposed. This greatly suppresses leakage output from the piezoelectric gyroscope and also improves stability. The operation in the drive mode non-resonant condition and detection mode resonance condition is also proposed. It is possible to increase the sensor sensitivity remarkably because the drive voltage can be increased. In addition, an equivalent circuit constant including the leakage output could be accurately deduced. Automatic compensation of temperature characteristic for angular velocity detection sensitivity and drift is examined with the simple equivalent circuit. By adding a new sensor that detects the drive vibrational mode, we can produce a stable reference signal for the phase detection of output. This shows that it is possible to greatly improve the temperature characteristic of sensitivity and drift. PMID- 10829627 TI - Optimum contact conditions for miniaturized surface acoustic wave linear motor AB - This paper reports the successful operation of a 70 MHz driving surface acoustic wave (SAW) linear motor with a miniaturized stator transducer. This paper also deals with an investigation into an optimized slider design for the miniaturized SAW linear motor. The performance of three silicon type sliders, with different projection size, was compared. Output forces of the three sliders were measured with change of pre-load. It was found that the slider with smaller projection tended to produce greater output force. PMID- 10829628 TI - Development of a self-oscillating ultrasonic micro-motor and its application to a watch AB - We have developed an ultrasonic micro-motor for use as a micro-actuator in place of an electromagnetic motor. This ultrasonic micro-motor, which can be driven by a single signal and in which the change of the direction of the rotor movement can be made easily by selecting the electrode to apply the driving signal, can easily construct a self-oscillating circuit and simplify the driving circuit. We have also simplified the motor structure, which is easy to miniaturize and mass produce. We applied a version of this motor with a diameter of 8 mm to a vibration alarm, and one with a diameter of 4.5 mm to a driving source of a calendar mechanism in a watch. This ultrasonic micro-motor is expected to be of use as a new driving source in a broad range of fields. PMID- 10829629 TI - Holding characteristics of planar objects suspended by near-field acoustic levitation AB - The authors have found the acoustic levitation phenomenon where planar objects of 10 kg weight can be levitated near a vibration surface. This phenomenon has been studied for non-contact transportation. A circular planar object can be suspended without contacting a circular vibration plate. We have studied the holding force which acts horizontally on the levitated objects. The horizontal position of the object is stabilized by this force. In this paper, we discuss the effect of the radius of a levitated object, levitation distance, displacement amplitude of the vibration plate and the vibration mode on the suspending force. PMID- 10829630 TI - Non-linear actuation of cantilevers using giant magnetostrictive thin films AB - In the present paper, we report the first results concerning a centimetre cantilever driven through non-linear effects in thin magnetostrictive films. Bimorph centimetre beams (24 mm x 5 mm) were designed using 160 microns thick glass and 0.4 micron thick multilayer TbFe/Fe films deposited via RF sputtering, using several sputtering conditions. Dynamic magneto-elastic excitations were studied experimentally on the few eigenresonance modes of the cantilevers. Sub harmonic excitation at half or one-third of the frequency of an eigenresonance mode has been achieved near the spin reorientation transition and the efficiency is as good as for eigenfrequency excitation. Specific magnetic properties of the samples led to a model of the thermodynamic potentials and behaviour of the excited resonator. PMID- 10829631 TI - Transverse and torsional complex vibration systems for ultrasonic seam welding of metal plates AB - Transverse and torsional complex vibration systems for ultrasonic seam welding of metal plate specimens, using a 27 kHz complex vibration disk welding tip vibrating in transverse and torsional vibration modes, were studied. Using a complex vibration welding system with a welding tip vibrating in elliptical or circular locus, thick plate specimens can be welded with a more uniform and larger area compared to a conventional ultrasonic welding system. The disk welding tip vibrates in an elliptical or circular locus. The complex vibration system can continuously weld multiple parts of metal plate specimens such as heat sinks with a large number of fins. PMID- 10829632 TI - One-dimensional longitudinal-torsional vibration converter with multiple diagonally slitted parts AB - For increasing the available vibration velocity of the one-dimensional longitudinal-torsional vibration converter, a new type of complex vibration converter with multiple slitted parts installed in the positions avoiding longitudinal nodal positions along the converter for decreasing the maximum vibration stress level at the vibration nodal part was studied. The free end of the converter vibrates in an elliptical or circular locus. Complex vibration systems with elliptical to circular or rectangular to square loci can be applied effectively for various high-power applications, including ultrasonic welding of metal or plastics, ultrasonic wire bonding of IC, LSI and electronic devices, and also ultrasonic motors. The converter with multiple slitted parts was improved in the vibration stress level and the quality factor compared with the converter with single slitted part. PMID- 10829633 TI - High-frequency ultrasonic wire bonding systems AB - The vibration characteristics of longitudinal-complex transverse vibration systems with multiple resonance frequencies of 350-980 kHz for ultrasonic wire bonding of IC, LSI or electronic devices were studied. The complex vibration systems can be applied for direct welding of semiconductor tips (face-down bonding, flip-chip bonding) and packaging of electronic devices. A longitudinal complex transverse vibration bonding system consists of a complex transverse vibration rod, two driving longitudinal transducers 7.0 mm in diameter and a transverse vibration welding tip. The vibration distributions along ceramic and stainless-steel welding tips were measured at up to 980 kHz. A high-frequency vibration system with a height of 20.7 mm and a weight of less than 15 g was obtained. PMID- 10829634 TI - Acoustic behaviour of current ultrasound contrast agents. AB - A general law gives the approximate change in signal level obtained in a particular imaging mode when a suitable contrast agent is added. It also shows that reduction of background signals is essential to overcome limitations found mainly in conventional (linear) ultrasound contrast imaging. Contrast agents contain stabilized microbubbles with very helpful non-linear properties. Acoustic methods for non-destructive and destructive testing of microbubbles are briefly discussed. In the main part, the linear and non-linear acoustic behaviour of various types of contrast agent are described. The latter is useful for new applications in diagnostic ultrasound. PMID- 10829635 TI - Detection procedures of ultrasound contrast agents. AB - In the early days, it was believed that ultrasound contrast agents (UCA) could be sufficiently detected and imaged with the conventional imaging methods nowadays referred to as fundamental imaging. Newer imaging techniques proved to be more sensitive and are based on specific properties of the UCA. In general, these new characteristics involve non-linear and transient characteristics of contrast agents that appear at the high end of the diagnostic acoustic intensity. Imaging modalities used today for UCA are, besides fundamental imaging, second harmonic imaging, power Doppler, harmonic power Doppler, pulse inversion and pulse inversion Doppler, multi-pulse imaging and subharmonic imaging. Although the results of conventional second harmonic imaging are still not optimal for perfusion imaging applications, in combination with Doppler techniques (colour Doppler, power Doppler) it is one of the most sensitive techniques currently available in terms of agent-to-tissue ratio. Further improvements in current and future detection methods demand a complete understanding of the ultrasound-UCA interaction. PMID- 10829636 TI - Subharmonic imaging of contrast agents. AB - Ultrasound contrast agents promise to improve the sensitivity and specificity of diagnostic ultrasound imaging. It is of great importance to adapt ultrasound equipment for optimal use with contrast agents e.g., by exploiting the nonlinear properties of the contrast microbubbles. Harmonic imaging is one technique that has been extensively studied and is commercially available. However, harmonic imaging is associated with problems, due to second harmonic generation and accumulation within the tissue itself. Given the lack of subharmonic generation in tissue, one alternative is the creation of subharmonic images by transmitting at the fundamental frequency (fo) and receiving at the subharmonic (fo/2). Subharmonic imaging should have a much better lateral resolution and may be suitable for scanning deep-lying structures owing to the higher transmit frequency and the much smaller attenuation of scattered subharmonic signals. In this paper, we will review different aspects of subharmonic imaging including implementation, in-vitro gray-scale imaging and subharmonic aided pressure estimation. PMID- 10829637 TI - The acoustics of diagnostic microbubbles: dissipative effects and heat deposition. AB - We discuss the effectively detectable scattered intensity of ultrasound from diagnostic microbubble suspensions, taking dissipative mechanisms in the liquid medium into account. In particular, we conclude that neither non-linear wave steepening of the incident (driving) wave nor of the outgoing (scattered) wave has a large effect on the scattered signal from typical bubbles. It is shown that, paradoxically, the far-field solution of the wave field is sufficient to compute the magnitude of expected temperature rises in the medium due to acoustic heat deposition, although appreciable heating is limited to intermediate-field distances from the bubble surface. PMID- 10829639 TI - Comparison of fundamental and wideband harmonic contrast imaging of liver tumors. AB - Wideband harmonic imaging (with phase inversion for improved tissue suppression) was compared to fundamental imaging in vivo. Four woodchucks with naturally occurring liver tumors were injected with Imagent (Alliance Pharmaceutical Corp., San Diego, CA). Randomized combinations of dose (0.05, 0.2 and 0.4 ml/kg) and acoustic output power (AO; 5, 25 and 63% or MI < or = 0.9) were imaged in gray scale using a Sonoline Elegra scanner (Siemens Medical Systems, Issaquah, WA). Tumor vascularity, conspicuity and contrast enhancement were rated by three independent observers. Imagent produced marked tumor enhancement and improved depiction of neovascularity at all dosages and AO settings in both modes. Tumor vascularity and enhancement correlated with mode, dose and AO (P < 0.002). Fundamental imaging produced more enhancement (P < 0.05), but tumor vascularity and conspicuity were best appreciated in harmonic mode (P < 0.05). Under the conditions studied here, the best approach was wideband harmonic imaging with 0.2 ml/kg of Imagent at an AO of 25%. PMID- 10829638 TI - Breast tumor vascularity identified by contrast enhanced ultrasound and pathology: initial results. AB - Quantifiable measures of vascularity obtained from contrast enhanced color flow images were correlated with pathologic vascularity measurements in ten female patients with a solid breast mass. Each patient received Levovist Injection (Berlex Laboratories Inc., Montville, NJ). Color flow images pre- and post contrast were obtained using an HDI 3000 unit (ATL, Bothell, WA) before removing the mass for pathologic vascularity assessments. Image-processing techniques were used to obtain both the ultrasound and pathologic vascularity measurements. Multiple linear regression found significant correlations for ultrasonic vascularity measurements post contrast and pathology (P = 0.02 and 0.06). No correlations were found between pre-contrast ultrasound and pathology. In conclusion, post-contrast ultrasonic flow measures provide a non-invasive measure of breast tumor neovascularity. However, the patient population is small, and until further patients are analyzed, these conclusions are preliminary. PMID- 10829640 TI - Simulation of electromechanical coupling coefficient by modified modal frequency spectrum method including the electrode effect AB - An approximate formula has been developed to determine the electromechanical coupling coefficient of the piezoelectric film in a four-layer composite resonator, which includes two electrodes. In this formula, the coupling coefficient can be calculated from the distribution of the effective coupling coefficients, k2eff, which are given by the modal frequency spectrum of the composite resonator. The feasibility of this method has been demonstrated with a ZnO/SiO2 composite resonator for different electrode thickness. The effect of mechanical propagation loss in both piezoelectric layer and substrate has also been studied. PMID- 10829641 TI - Transducer transmitting sensitivity measurements in restricted environments AB - Transducer transmitting sensitivity is normally measured under free-field conditions. In laboratory tanks, this is often achieved by using time gating to isolate the direct signal from the reflected signals. However for high-Q, low frequency projectors, the free-time availability in laboratory tanks may be too short. A measurement technique has been developed to perform these calibration measurements in the presence of a reverberant field. The measurements have been performed at ultrasonic frequencies so that the free-field sensitivity can be determined in order to assess the accuracy of this new technique. The technique involves extracting the direct field from the combination of the direct and reverberant fields by plotting the variation of pressure squared against the reciprocal of separation squared. The gradient of this graph is proportional to the acoustic power radiated into the tank, and from this, the direct field pressure and projector sensitivity can be calculated. This paper investigates this technique and looks at the influences of tank size and construction, as well as the effect of averaging over different bandwidths. PMID- 10829642 TI - Measuring vibration characteristics at large amplitude region of materials for high power ultrasonic vibration system AB - This study proposes a method of estimating the mechanical quality factor of materials for high-power ultrasonic vibration systems under large vibration amplitude conditions. The quality factors of several metals as well as some polymers are measured by this method. In this method, the quality factor is calculated as the ratio of the reactive energy stored in a specimen to the dissipated energy. The dissipated energy is estimated from the input/output mechanical energy to the specimen by measurement of the vibration intensity, while the reactive energy is measured as the kinetic energy of the vibration. Then, the quality factor for the specified part can be extracted using this method. In this report, quality factors for torsional vibration were measured at about 30 kHz as functions of the vibration strain. PMID- 10829644 TI - Ultrasonic non-destructive testing of pieces of complex geometry with a flexible phased array transducer AB - Ultrasonic non-destructive testing of components of complex geometry in the nuclear industry faces several difficulties: sensitivity variations due to unmatched contact, inaccurate localization of defects due to variations of transducer orientation, and uncovered area of the component. To improve the performances of such testing and defect characterization, we propose a new concept of ultrasonic contact phased array transducer. The phased array transducer has a flexible radiating surface able to fit the actual surface of the piece to optimize the contact and thus the sensitivity of the test. To control the transmitted field, and therefore to improve the defect characterization, a delay law optimizing algorithm is developed. To assess the capability of such a transducer, the Champ-Sons model, developed at the French Atomic Energy Commission for predicting field radiated by arbitrary transducers into pieces, has to be extended to sources directly in contact with pieces of complex geometry. The good behavior of this new type of probe predicted by computations is experimentally validated with a jointed transducer positioned on pieces of various profiles. PMID- 10829643 TI - The application of high permittivity piezoelectric ceramics to 2D array transducers for medical imaging. AB - Two-dimensional (2D) array transducers have become of great interest in the last few years, in view of real-time volumetric ultrasonic imaging. The electrical matching between the high electrical impedance of elements and the standard cables and electronics is one of the key issues in 2D array design. The use of high-permittivity ceramics such as PNNZT either in bulk configuration or in 1-3 piezocomposites decreases the electrical impedance. In this paper, bulk samples of PNNZT and PZT ceramics are characterised, and results are compared. 2D array elements are then manufactured and their electrical impedances measured. Theoretical predictions of homogenisation models for 1-3 piezocomposites allow the simulation of the electroacoustic behaviour of 2D array elements. Results for both piezocomposite and bulk materials can be obtained. Calculations of the input impedance, the sensitivity and the bandwidth of the different configurations are compared and discussed. These results demonstrate the advantages of the PNNZT compositions over standard PZT. PMID- 10829645 TI - Sensitivity of ultrasonic hydrophone probes below 1 MHz. AB - Frequency responses of different PVDF polymer hydrophone probes, including membrane and needle designs, were measured and are presented in terms of end-of cable voltage sensitivity versus frequency over a wide, 4.5 octave bandwidth ranging from 0.25 to 2.5 MHz. The probes are seldom, if at all, characterized in this frequency range due to the difficulties associated with a lack of adequate and readily implementable calibration techniques. To this end, a technique, which uses a combination of swept frequency chirp and reciprocity, so that both the relative and absolute plots of sensitivity versus frequency can be obtained, was developed and tested. Salient features of the technique including the design of a 6 octave auxiliary acoustic source are described. The experimental data indicate that a majority of the PVDF membrane hydrophones exhibit a relatively uniform (to within +/- 2 dB) response. While, in general, this is not the case for commercially available needle hydrophone probes, it is evidenced that a careful attention to the PVDF probe design results in frequency characteristics fairly close to those achievable with a membrane design. The overall uncertainty of the calibration technique was estimated to be better than +/- 1.5 dB in the considered frequency range. The results of this work are important to implement procedures for adequate determination of the Mechanical Index (MI) of ultrasound imaging devices. MI is widely accepted as a predictor of potential bioeffects associated with cavitation phenomena. Current efforts are focused on extending the applicability of the technique to frequencies below 100 kHz. PMID- 10829646 TI - Definition and measurement of the normalized electrical impedance of lossy piezoelectric resonators for ultrasonic transducers AB - Relevant equivalent circuit parameters and values of material constants of a piezoelectric resonator can be determined from measurements of its electrical input impedance as a function of frequency. The complex electrical impedance curves and the associated critical frequencies are the basis of this characterization by the piezoelectric resonance method. In this paper, the previously introduced concept of normalized electrical impedance of the lossy resonator, extended to include piezoelectric losses, is applied to the analysis of the effects of different types of intrinsic losses on peak values, bandwidths and characteristic frequencies. The resulting impedance patterns depend solely on the electromechanical coupling coefficient and the loss tangents, providing a useful tool for the analysis of low-Q resonators. The normalized impedance is experimentally evaluated from the basic data provided by an HP 4194A impedance analyser by means of specifically developed ASP programs. PMID- 10829647 TI - Transducer resolution enhancement by combining different excitation pulses AB - This work presents a low-cost solution to improve the axial resolution of an arbitrarily loaded and backed piezoelectric transducer. The proposed solution is based on the use of different excitation pulses and, in particular, the combination of two excitation pulses. The characteristic parameters of the second pulse, i.e. amplitude and time delay with respect to the main pulse, are obtained by studying the waveform of a reference pulse emitted or received by the transducer. This reference pulse is the transducer's response to a generic excitation pulse which has the same shape and duration as the main pulse. The work also presents a procedure to estimate the characteristic parameters of the second excitation pulse by means of the electrical simulator SPICE. Then, to illustrate the effectiveness of the simulation, the simulated and experimental values are compared through two experimental cases. PMID- 10829648 TI - Nonlinear mechanical behavior of piezocomposites for ultrasonic transducers AB - A comparative study is carried out between the nonlinear behavior of a composite and the piezoceramic used to obtain it. This characterization is necessary for using the composite in power transducer applications. A study of the losses and the resonator stiffness variations has also been done. Both these effects, as well as the possibility of the frequency hysteresis, show different behavior in the composites, since the increases in the ceramics are different from those in the composites. In this study two measurement methods are used: principally the motional impedance increase with the motional current measurements. The results obtained are normalized in order to make them independent of the resonator size, and thus make the comparison between the composite and the ceramic easier. The figure of the mechanical loss tangent tan delta m versus the mean strain shows that the losses can be greater in the ceramic than in the composite for soft ceramics. The dependence behavior of the losses and stiffness variations versus the mean strain is studied for both resonators. PMID- 10829649 TI - A new beamforming technique for ultrasonic imaging systems AB - We propose a simple, versatile and inexpensive beamforming method that performs the aperture windowing of an ultrasonic transducer array in the transmit mode, without modifying the driver voltage, but simply controlling the length of the electric pulse driving the array elements. A conversion formula has been determined that permits us to compute, for a desired emitted pulse amplitude, the corresponding driving pulse length to be applied. Any shading function can be implemented over any type of transducer array, using very low-cost hardware. Computer simulations and experimental measurements, with a 3.8 MHz convex array, confirm the effectiveness of this approach in enhancing the contrast resolution, since the off-axis intensity in the radiated beam pattern is largely reduced. PMID- 10829650 TI - Aperture and element minimization in linear sparse arrays with desired beam patterns AB - In this paper, an optimization method aimed at designing aperiodic linear sparse arrays is proposed that is based on the stochastic optimization methodology called simulated annealing. Such a method optimizes the positions and the weight coefficients of each element of a linear array in order to obtain a beam pattern that meets given requirements. Moreover, while positions and weights are optimized, minimization of both the number of elements and the spatial aperture is carried out. In this way, a beam pattern without grating lobes and close to the desired one is produced using a small number of elements and a short aperture. The results obtained are impressive in terms of improvement of array characteristics and performance over those reported in the literature, while a great flexibility in defining the desired beam pattern is allowed. Finally, it is proved that the array configurations need a number of elements very close to the absolute minimum to achieve the desired beam pattern. PMID- 10829651 TI - Synthetic aperture imaging by scanning acoustic microscopy with vector contrast AB - Based on phase sensitive scanning acoustic microscopy (PSAM), a novel scheme suitable for volume imaging has been developed. The method employs synthetic aperture insonification combined with synthetic aperture imaging. Excitation and detection are performed by planar scanning of two focusing transducer and vector (phase and amplitude) detectors for the ultrasonic wave packages observed in transmission. Examples for applications of the scheme, including technically relevant simplifications based on reduced dimensions of the scan ranges, are presented. Detection schemes already applied for non-destructive testing (NDT) and non-destructive evaluation (NDE) of the mechanical properties of functionally graded materials are an example for the application of the generalised approach presented here with reduced dimensions of the scan. The technique is suitable for NDT and NDE imaging respectively with three-dimensional resolution. PMID- 10829652 TI - Application of orthogonal ultrasonic signals and binaural processing for imaging of the environment AB - Data acquisition rates in ultrasonic imaging systems are limited by the finite value of the speed of ultrasonic waves. In order to improve the imaging speed, it is proposed to perform simultaneous scanning of the environment in different directions. In order to avoid cross-talk between adjacent channels in different directions, different orthogonal signals are transmitted. Application of cross correlation processing and non-linear iterative deconvolution enables the reliable separation of signals transmitted by different sources and reflected by multiple targets. The spatial positions of the targets are found using the data obtained after the non-linear deconvolution as the initial data for binaural or tri-aural processing. This approach has been exploited in ultrasonic sonar used for navigation of mobile robots. PMID- 10829653 TI - Incidence of element distribution on the ultrasonic field of segmented annular arrays AB - Segmented annular arrays are sometimes used for 3D ultrasonic imaging. However, owing to their geometrical complexity, the acoustic field generated by this type of aperture has not been adequately described. In this work, a method based on the array factor approach is used to describe the field radiated by sector annular arrays. This approach allows one to analyse the influence on the field of several aperture parameters, such as the number of elements per annulus, size and spatial distribution of elements, etc. In addition, strategies to reduce grating lobes are presented. PMID- 10829654 TI - A multirate scan conversion method AB - B-mode ultrasonic imaging requires that the acquired polar coordinate ultrasound data be converted to the Cartesian format used by digital monitors. Image quality depends on the interpolation algorithm used to this purpose. In this work a selective sampling technique, based on acquiring data at specific points of the scanned area together with a straightforward linear interpolation step, is proposed. Hardware complexity is avoided, because the interpolation task can be carried out by software in real time, concurrently with data acquisition. The performances of the proposed approach are analysed with regard to those provided by other algorithms and some implementation issues are addressed. PMID- 10829655 TI - Potential of coded excitation in medical ultrasound imaging. AB - Improvement in signal-to-noise ratio (SNR) and/or penetration depth can be achieved in medical ultrasound by using long coded waveforms, in a similar manner as in radars or sonars. However, the time-bandwidth product (TB) improvement, and thereby SNR improvement is considerably lower in medical ultrasound, due to the lower available bandwidth. There is still space for about 20 dB improvement in the SNR, which will yield a penetration depth up to 20 cm at 5 MHz [M. O'Donnell, IEEE Trans. Ultrason. Ferroelectr. Freq. Contr., 39(3) (1992) 341]. The limited TB additionally yields unacceptably high range sidelobes. However, the frequency weighting from the ultrasonic transducer's bandwidth, although suboptimal, can be beneficial in sidelobe reduction. The purpose of this study is an experimental evaluation of the above considerations in a coded excitation ultrasound system. A coded excitation system based on a modified commercial scanner is presented. A predistorted FM signal is proposed in order to keep the resulting range sidelobes at acceptably low levels. The effect of the transducer is taken into account in the design of the compression filter. Intensity levels have been considered and simulations on the expected improvement in SNR are also presented. Images of a wire phantom and clinical images have been taken with the coded system. The images show a significant improvement in penetration depth and they preserve both axial resolution and contrast. PMID- 10829656 TI - Decomposition of acoustic fields in quantised Bessel beams AB - Bessel beams are non-diffracting solutions to the wave equation, which become limited diffraction beams when implemented on finite apertures. In previous work we have applied Fourier-Bessel theory to deduce theoretically that the quantization of the Bessel beam profile on annular arrays results in a field which is a sum of limited diffraction fields. Here we demonstrate this for a five ring, 20-mm diameter, 2.5-MHz transducer. The quantized field comprises a weighted sum of a main component corresponding to the desired field, along with three other major components and 28 lesser components representing undesired field components. The three major components correspond to limited diffraction beams with narrower beamwidths and shorter depths of field than the desired beam, and we show that these account for most of the discrepancies between the desired field and actual quantized field. An estimate and an interpretation of the number of field components as a function of the wavenumber are also given. PMID- 10829657 TI - A frequency domain inversion method applied to propagation models in unconsolidated granular materials AB - The measurement setup consists of a water filled tank within which a smaller Plexiglas tank containing a known sediment is placed. This provides a Plexiglas sediment-Plexiglas configuration, which is then used in conjunction with transducers and a pulser-receiver for a transmission and reflection experiment. The aim of this work is to find some physical parameters of the sediment from these measurements. A general viscoelastic model that considers losses due to absorption and dispersion along the propagation of the wave through the sediments is proposed. A rational transfer function is used to model the viscoelastic modulus of the bulk sediments, and comparisons with constant Q viscoelastic modelling and Biot modelling are performed. The estimation of the model parameters is elaborated using a maximum likelihood estimator in the frequency domain. PMID- 10829658 TI - Ultrasonic characterization of liquids using resonance antireflection AB - This paper presents an ultrasonic method for measuring the density of liquids with a solid layer separating a reference fluid and a test fluid. By adjusting the frequency of the exciting signal according to the thickness of the layer, it is possible to generate destructive interference of the waves reflected at the first and at the second boundary of the layer. Thus, the layer appears to vanish for the incident waves. The resulting echo signal depends only on the acoustic impedances of the reference fluid and the test fluid and the density which is of interest can be extracted. Short and long-term drifts of the electronics and the ultrasonic transducer implied are eliminated by using the well-known pulse-echo technique with additional frontwave detection. PMID- 10829659 TI - Resonant frequency method for the measurement and uncertainty analysis of acoustic and elastic properties AB - A resonant frequency method is proposed to determine the bar speed and the Poisson's ratio of a rod several meters long of the kind often used for manufacturing power sonic transducers and mechanical horns in mass production. This method is shown to be robust in a field test and with high resolution for estimation of the bar speed. The uncertainty (standard deviation/average) for the bar speed and the Poisson's ratio can be reduced by appropriately selecting the mode number pairs and increasing the mode number. In this study, the uncertainties of the bar speed and Poisson's ratio are less than 0.043% and 2.9%, respectively. The resonant frequency method is verified by comparing the dilatational wave speeds calculated by elastic theory with those from experiments, the difference between them being less than 0.17%. PMID- 10829660 TI - Surface and friction characterization by thermoelectric measurements during ultrasonic friction processes AB - Even though friction is one of the oldest problems in physics many aspects of friction processes are not clear today. We present an experimental setup, which permits the study of tribological systems by measuring the dissipated heat at the interface of two surfaces during a friction process with a time resolution of 1 ms. The apparatus is based on a standard ultrasonic wire-bond machine used in semiconductor industries to connect the internal semiconductor die to the external leads, but the standard bond wire is replaced by a thermocouple. To demonstrate the ability of the apparatus it will be shown that bond substrates used in semiconductor industries can be unequivocally characterized. PMID- 10829661 TI - Ultrasonic early detection of the initiation of a ductile tearing at very low temperature AB - The possibility of performing early ultrasonic detection of the initiation of ductile tearing in a three point bending test specimen had been demonstrated for temperatures between room temperature and -140 degrees C. PMID- 10829662 TI - Comparison of several methods to characterise the high frequency behaviour of piezoelectric ceramics for transducer applications AB - Thickness mode resonances in commercial piezoelectric ceramics have been characterised as a function of frequency by two methods. The first is based on a fit on the electrical impedance for the fundamental and the overtones. This method has been applied to a large number of PZT ceramic samples and frequency dependence for all the parameters is investigated, in particular for the piezoelectric coefficient e33. The second is based on the measurement of the mechanical displacement at the centre of the surface of a PZT ceramic disk. With a modified KLM scheme, this displacement is modelled. The dielectric, elastic and piezoelectric parameters are extracted and compared for the fundamental and the third overtone. The results are found to be in good agreement. PMID- 10829663 TI - Ultrasonic characterization of materials hardness AB - In this paper, an experimental technique has been developed to measure velocities and attenuation of ultrasonic waves through a steel with a variable hardness. A correlation between ultrasonic measurements and steel hardness was investigated. PMID- 10829664 TI - Elastic waves in fibre composite laminates AB - This paper considers the propagation of elastic waves in an eight-ply quasi isotropic laminate arising from line sources of dislocation located at each of the seven interfaces in turn. The line source sets up a straight crested wave travelling along the laminate in a direction normal to the load line, and the elastodynamic equations within each layer are solved by taking the Laplace transform with respect to time and the Fourier transform with respect to the spatial coordinate in the direction of propagation. The resulting system of six first-order differential equations in each layer is solved to obtain the transforms of the displacement and stress components throughout the laminate. The time history of any displacement or stress component at any location may then be recovered by numerical inversion of the double transform. Examples are shown of the time history of the normal displacement of the top surface of the laminate at a distance of 20 plate thicknesses from the plane of action of the sources. The numerical inversion involves a summation over different modes of Rayleigh-Lamb waves in the laminate and contributions to the overall response from some of the individual modes are displayed. PMID- 10829665 TI - Angular spectrum approach for the computation of group and phase velocity surfaces of acoustic waves in anisotropic materials AB - The decomposition of an acoustic wave into its angular spectrum representation creates an effective base for the calculation of wave propagation effects in anisotropic media. In this method, the distribution of acoustic fields is calculated in arbitrary planes from the superposition of the planar components with proper phase shifts. These phase shifts depend on the ratio of the distance between the planes to the normal component of the phase slowness vector. In anisotropic media, the phase shifts depend additionally on the changes of the slowness with respect to the direction of the propagation vector and the polarization. Those relations are obtained from the Christoffel equation. The method employing the fast Fourier transformation algorithm is especially suited for volume imaging in anisotropic media, based on holographic detection in transmission of acoustic waves generated by a point source. This technique is compared with measurements on crystals performed by phase-sensitive scanning acoustic microscopy. PMID- 10829666 TI - 3-D voltage model for detection of sound radiated from anisotropic materials AB - The elastic behavior of composite materials has been characterized experimentally by employing a 3-D voltage calculation to model transmission or reflection experiments. With sound propagation along material symmetry directions, integration over the incident-plane angle alone is generally sufficient to model the transducer voltage accurately. In general material directions this integration must be extended to account for asymmetrical variations in the reflection or transmission coefficient out of the incident plane. Theoretical and experimental results illustrate this effect and the relationship between 2-D and 3-D calculations. Experimental measurements are used to in the reconstruction of viscoelastic properties in composite plates. The influence of the phi-dependent integration on the voltage, in the 3-D calculation, is particularly strong when the incident angles are small and the wave paths are large, as typically experienced in air-coupled measurements. PMID- 10829667 TI - Ultrasonic measurement of elastic constants in fiber-reinforced polymer composites under influence of absorbed moisture AB - This paper presents an attempt to quantify hygral aging in fiber-reinforced polymer composites by the elastic constants C11 and C33. Quantitative ultrasonic measurements of the elastic constants for three different unidirectional as well as three different cross-ply specimens were compared. The specimens were manufactured with different moisture resistant surfaces and immersed in water for 24 h. By calculating the elastic constants, it was taken into account that hygral aging was accompanied by absorption of moisture in the polymer matrix. Moisture changed the laminate dimensions significantly, and typically moisture expansion coefficients are reported. Moreover, as the ultrasonic pulse form changed in the anisotropic materials, different broadband methods were used to calculate the elastic constants. PMID- 10829668 TI - Ultrasonics and X-ray diffraction under pressure in the Paris-Edinburgh cell AB - Our objective consists in validating a new set-up which will permit us to carry out simultaneously ultrasonic and X-ray diffraction measurements under pressure. To validate the results obtained by this new set-up, the elastic properties of a single crystal of germanium were studied. Our results are in good agreement with those of Goncharova et al. and McSkimin and Andreatch. The results of the present study are compared with those of Menoni et al. and obtained by X-ray diffraction in a diamond anvil cell. PMID- 10829669 TI - Wave propagation in an anisotropic nickel-based superalloy AB - The effects of elastic anisotropy on ultrasound propagation in a nickel-based single crystal test component are studied using a 25 MHz focused probe in a water immersion system. Anisotropy gives rise to directionally dependent acoustic wavespeeds, beam steering, acoustic energy focusing and mode conversion for normal incidence. Transverse mode echoes are particularly strong in the vicinity of crystallographic directions in which the Gaussian curvature of the slowness surface is zero and divergence of the echo amplitude is predicted on the basis of the stationary phase approximation. There are other directions where the transverse mode echoes vanish for symmetry reasons. The longitudinal mode echo amplitude also shows significant variation with direction. Overall there is good agreement between the echo signal arrival times and amplitudes we measure and calculation. Progress in applying this technique to gas turbine blades is reported. PMID- 10829670 TI - The propagation of ultrasound in an austenitic weld AB - The propagation of ultrasound through an austenitic weld is investigated experimentally as well as in a numerical simulation. The weld is insonified at normal incidence to the fusion line with a longitudinal contact transducer. In order to experimentally trace the ultrasound through the weld, slices of different thicknesses from the original weld have been fabricated. Through transmission A-scans have then been produced for each weld slice and compared with the corresponding numerical simulation. A comparison of the direction of ultrasound propagation through the weld for the two approaches shows quite good agreement. However, attenuation due to scattering at grain boundaries in the weld is poorly modelled in the simulation. In order to improve this, a better model of the weld is needed. PMID- 10829671 TI - A computational method for wave propagation from a point load in an anisotropic material AB - One approach which is employed to solve dynamic point load problems in plates and laminates is to take integral transforms to reduce the governing equations to a system of ordinary differential equations with respect to the depth variable. The solution of this system leads to expressions for the transforms of the displacement and stress components at any level in the plate and the transient response at any location may then be recovered by inversion of the multiple transforms. The formal transform inversion involves a double infinite integral but by making a change of variable this may be replaced by an infinite integral associated with a line source and a finite integral with respect to the orientation of the line. A first attempt at applying this approach to obtain the point load response of quasi-isotropic fibre composite laminate led to a non causal predicted signal. This paper deals with an investigation of this proposed method applied to the simpler model problem of wave propagation in a two dimensional anisotropic medium. Results are obtained for two different time histories of point loads, namely: a delta function; and a single period of a sine function. In the case of the delta function source a comparison is made with the analytic solution and the errors arising from the numerical approach are discussed. Graphs are also presented showing the non-causal contributions to the overall response which arise at individual angles of orientation of the line source. PMID- 10829672 TI - Nonlinear propagation in ultrasonic fields: measurements, modelling and harmonic imaging. AB - In high amplitude ultrasonic fields, such as those used in medical ultrasound, nonlinear propagation can result in waveform distortion and the generation of harmonics of the initial frequency. In the nearfield of a transducer this process is complicated by diffraction effects associated with the source. The results of a programme to study the nonlinear propagation in the fields of circular, focused and rectangular transducers are described, and comparisons made with numerical predictions obtained using a finite difference solution to the Khokhlov Zabolotskaya-Kuznetsov (or KZK) equation. These results are extended to consider nonlinear propagation in tissue-like media and the implications for ultrasonic measurements and ultrasonic heating are discussed. The narrower beamwidths and reduced side-lobe levels of the harmonic beams are illustrated and the use of harmonics to form diagnostic images with improved resolution is described. PMID- 10829674 TI - On some nonlinear effects in ultrasonic fields AB - Nonlinear effects associated with intense sound fields in fluids are considered theoretically. Special attention is directed to the study of higher effects that cannot be described within the standard propagation models of nonlinear acoustics (the KZK and Burgers equations). The analysis is based on the fundamental equations of motion for a thermoviscous fluid, for which thermal equations of state exist. Model equations are derived and used to analyze nonlinear sources for generation of flow and heat, and other changes in the ambient state of the fluid. Fluctuations in the coefficients of viscosity and thermal conductivity caused by the sound field, are accounted for. Also considered are nonlinear effects induced in the fluid by flexural vibrations. The intensity and absorption of finite amplitude sound waves are calculated, and related to the sources for generation of higher order effects. PMID- 10829673 TI - Finite difference modelling of the temperature rise in non-linear medical ultrasound fields. AB - Non-linear propagation of ultrasound can lead to increased heat generation in medical diagnostic imaging due to the preferential absorption of harmonics of the original frequency. A numerical model has been developed and tested that is capable of predicting the temperature rise due to a high amplitude ultrasound field. The acoustic field is modelled using a numerical solution to the Khokhlov Zabolotskaya-Kuznetsov (KZK) equation, known as the Bergen Code, which is implemented in cylindrical symmetric form. A finite difference representation of the thermal equations is used to calculate the resulting temperature rises. The model allows for the inclusion of a number of layers of tissue with different acoustic and thermal properties and accounts for the effects of non-linear propagation, direct heating by the transducer, thermal diffusion and perfusion in different tissues. The effect of temperature-dependent skin perfusion and variation in background temperature between the skin and deeper layers of the body are included. The model has been tested against analytic solutions for simple configurations and then used to estimate temperature rises in realistic obstetric situations. A pulsed 3 MHz transducer operating with an average acoustic power of 200 mW leads to a maximum steady state temperature rise inside the foetus of 1.25 degrees C compared with a 0.6 degree C rise for the same transmitted power under linear propagation conditions. The largest temperature rise occurs at the skin surface, with the temperature rise at the foetus limited to less than 2 degrees C for the range of conditions considered. PMID- 10829675 TI - A novel phase locked cavity resonator for B/A measurements in fluids AB - A new technique for the measurement in fluids of the acoustic non-linearity parameter B/A is presented, together with measured B/A values for several fluids. The non-linearity parameter is measured by phase locking radial modes within a PZT cylinder. The system, which implements the isentropic phase technique, uses continuous wave phase locking to measure the change in sound velocity that is typically associated with a change in ambient pressure under constant entropy. The method provides a means of measuring B/A in vitro both accurately and simply without the typical problems involved in time-of-flight systems. Fluid samples can remain small due to the nature of the cavity resonator, so the system is well suited to small volume, biological samples. PMID- 10829676 TI - Harmonic propagation of finite amplitude sound beams: experimental determination of the nonlinearity parameter B/A AB - We propose a method to determine the nonlinearity parameter B/A of a liquid from the spatial evolution of harmonic components. We describe an analytical model, in the parabolic and quasi-linear approximations, that predicts the continuous finite amplitude sound beam propagation radiated by a plane piston source. This model takes into consideration attenuation, diffraction and nonlinear effects. The fundamental and second harmonic ultrasonic fields are expressed as the superposition of Gaussian beams. Axial propagation curves are then compared with those obtained by direct numerical solution of the transformed beam equation using the finite difference method, and with experimental results. Accurate measurements of pressure levels for the nonlinearly generated harmonics in water are performed along and across the propagation axis for different pressure values delivered at the piston surface. Experimental results, for water and ethanol, are in agreement with those of our model, which allows us to obtain the expected value of the nonlinearity parameter B/A. PMID- 10829677 TI - Fluid dynamics phenomena induced by power ultrasounds AB - Propagation of power ultrasound (from 20 to 800 kHz) through a liquid inside a cylindrical reactor initiates acoustic cavitation and also fluid dynamics phenomena such as free surface deformation, convection, acoustic streaming, etc. Mathematical modelling is performed as a new approach to predict where active bubbles are and how intense cavitation is. A calculation based on fluid dynamics equations is undertaken using computational fluid dynamics code; this is of great interest because such code provides not only the pressure field but also velocity and temperature fields. The link between the acoustic pressure and the cavitation field is clearly established. Moreover, the pressure profile near a free surface allows one to predict the shape of the acoustic fountain. The influence of the acoustic fountain on the wave propagation is shown to be important. The convective flow inside a reactor is numerically obtained and agrees well with particle image velocity measurements. Non-linearities arising from the dissipation of the acoustic wave are computed and lead to the calculation of the acoustic streaming. The superimposed velocity field (convective flow and acoustic streaming) succeeds in simulating the bubble behaviour at 500 kHz, for instance. PMID- 10829678 TI - Measurement of the acoustic nonlinearity parameter in 1-alkanols AB - We present a harmonic generation method based on a pitch-catch experimental setup and the calculation of the corresponding nonlinear parameter in 1-alkanols, from methanol to 1-decanol. By focusing a 14 MHz ultrasonic toneburst through a spherical quartz lens in these alcohols, a second harmonic is produced and measured as a function of distance by a 28 MHz piezoelectric transducer placed near the focus. By extracting the point where the second harmonic power is at a maximum in each liquid, the corresponding nonlinear parameter is then computed with a model developed by Germain et al. The preliminary results from this experiment are in reasonable agreement with the values found in the literature for primary alcohols and support the idea that the nonlinear parameter is dependent on the chain structure. PMID- 10829680 TI - Frequency dependence of H2O2 generation from distilled water AB - The frequency dependence of H2O2 generation from H2O by a sonochemical reaction was detected experimentally. The results are in good agreement with previous experimental results, which indicate that in sonochemical reactions, frequencies higher than 90 kHz are more effective than frequencies of several tens of kilohertz. The phonon concept of acoustic waves makes it clear that energy depends on frequency, i.e. on the condition of equal phonon density; higher frequency means higher energy. The concentration and accumulation of acoustic energy will be performed through a bubble surface. From the analogy of photoelectric effects, the frequency dependence of the sonochemical reaction was discussed using the phonon concept. PMID- 10829679 TI - Time-domain modeling of nonlinear distortion of pulsed finite amplitude sound beams. AB - This work aims to validate a time domain numerical model for the nonlinear propagation of a short pulse of finite amplitude sound beam propagation in a tissue-mimicking liquid. The complete evolution equation is simply derived by a superposition of elementary operators corresponding to the 'one effect equation'. Diffraction LD, absorption and dispersion LAD, and nonlinear distortion LNL effects are treated independently using a first order operator-splitting algorithm. Using the method of fractional steps, the normal particle velocity and the acoustical pressure are calculated plane by plane, at each point of a two dimensional spatial grid, from the surface of the plane circular transducer to a specified distance. The LA operator is a time convolution between the particle velocity and the causal attenuation filter built after the Kramers-Kroning relations. The LNL operator is a time-based transformation obtained by following an implicit Poisson analytic solution. The LD operator is the usual Rayleigh integral. We present a comparison between theoretical and experimental temporal pressure waveform and axial pressure curves for fundamental (2.25 MHz), second, third and fourth harmonics, obtained after spectral analysis. PMID- 10829681 TI - Assessment of the adhesion quality of fusion-welded silicon wafers with nonlinear ultrasound AB - Diffusion bonded silicon wafers are employed in the semiconductor industry. Their bonding quality must be monitored by a nondestructive testing technique. We present an ultrasonic technique allowing us to monitor the quality of the diffusion bond by measuring the anharmonic content of a transmitted ultrasonic wave. The anharmonicity is caused by weak bonds and manifests itself at high dynamic strains exerted by the ultrasonic wave. The source of nonlinearity is located in the rim of delaminations in the interface. PMID- 10829682 TI - Acoustic nonlinearities in adhesive joints AB - Nondestructive testing of adhesive joints is of great interest. The method of second harmonic generation promises to give early information about failure of adhesive layers. In the case of resonance the amplitude of strain in a soft interface layer is strongly increased and, therefore, the layer considerably contributes to A2, the amplitude of the second harmonic. The nonlinear behavior of such a layer and its influence on A2 was studied by means of the finite element method. In the experimental situation all materials along the sound path contribute to A2. The dependence of the calculated and measured effects on the layer thickness, the velocity of sound, and the nonlinearity coefficient beta are reported. PMID- 10829683 TI - Observation of ultrasound velocity gradient in fullerene ceramics by acoustic microscopy AB - A scanning acoustic microscope is used to study the distribution of elastic properties in small samples (O 3 x 2 mm3) of new hard phases of C60. The specimens under investigation were synthesized from pure C60 powder under pressure P = 8 GPa in the temperature range 500-1650 K. The time-of-flight mode was used for bulk sound wave velocity determination in a direction parallel to the cylinder's axis. Longitudinal sound wave velocities greater than 10,000 m/s were found for all specimens treated at temperatures higher than 1000 K. Using the B-scan mode allowed us to observe the velocity gradient in the sample's periphery. The heterogeneous internal structure of the specimen is visualized in the images formed in C- and B-scan modes. PMID- 10829684 TI - A macrosonic system for industrial processing AB - The development of high-power applications of sonic and ultrasonic energy in industrial processing requires a great variety of practical systems with characteristics which are dependent on the effect to be exploited. Nevertheless, the majority of systems are basically constituted of a treatment chamber and one or several transducers coupled to it. Therefore, the feasibility of the application mainly depends on the efficiency of the transducer-chamber system. This paper deals with a macrosonic system which is essentially constituted of a high-power transducer with a double stepped-plate radiator coupled to a chamber of square section. The radiator, which has a rectangular shape, is placed on one face of the chamber in order to drive the inside fluid volume. The stepped profile of the radiator allows a piston-like radiation to be obtained. The radiation from the back face of the radiator is also applied to the chamber by using adequate reflectors. Transducer-chamber systems for sonic and ultrasonic frequencies have been developed with power capacities up to about 5 kW for the treatment of fluid volumes of several cubic meters. The characteristics of these systems are presented in this paper. PMID- 10829685 TI - Numerical modeling of high-power ultrasonic systems: current status and future trends AB - Numerical models of high power ultrasonic systems are usually based on finite element or boundary element methods. The basic physical models are linear and rely upon the theory of elasticity, the constitutive law of piezoelectricity and the theory of linear acoustics. They are only valid at low drive level. Some recent developments include nonlinearities of the transduction mechanism and of the propagation medium. In this paper, standard and advanced numerical models are discussed and illustrated by several application examples of high power ultrasonics. PMID- 10829686 TI - Application of ultrasound in comminution AB - This paper deals with a new technology for fine grinding of hard materials, based on a high-compression roller mill with ultrasonic capabilities. The machine was tested by producing fine powders from hard rocks, with and without ultrasonic activation, permitting the beneficial effects of ultrasound to be evaluated. The experimental set-up allows the following operational parameters to be measured: material flow, applied torque, angular velocity of the rollers, stress on the shafts, ultrasonic energy applied and the vibration amplitude and phase behaviour of the transducer roller. It is found that the application of ultrasonic energy diminishes the torque required, the stress over the shafts and the total energy consumed for the same grinding results. In addition, a reduction in the erosion of the grinding surfaces was found. The optimal value of the applied ultrasonic power was determined by measuring the specific rate of breakage, a parameter that refers to the energy consumed for the generation of 1 ton of material, for each size range. PMID- 10829687 TI - Environmental implications of acoustic aerosol agglomeration AB - An overview is presented of acoustically induced agglomeration of fine particulates, potential industrial applications of the technology, and its environmental implications in terms of fine particle pollutants. Adverse health effects due to exposure to fine aerosols are discussed as well as recent legislation to reduce the output of such emissions. Based on this, the need for new, more efficient particle filtration technologies is demonstrated. It is shown that acoustic aerosol preconditioning meets all the requirements to reduce the fine particle output of conventional filter systems. The results of laboratory scale experiments are presented to illustrate the underlying mechanisms of the acoustic agglomeration process, while data from pilot scale testing are shown to prove the effectiveness of acoustic agglomeration systems in reducing the fine particle content of aerosols. Other filtration technologies are compared with acoustic agglomeration equipment. PMID- 10829688 TI - Algorithms for estimating blood velocities using ultrasound. AB - Ultrasound has been used intensively for the last 15 years for studying the hemodynamics of the human body. Systems for determining both the velocity distribution at one point of interest (spectral systems) and for displaying a map of velocity in real time have been constructed. A number of schemes have been developed for performing the estimation, and the various approaches are described. The current systems only display the velocity along the ultrasound beam direction and a velocity transverse to the beam is not detected. This is a major problem in these systems, since most blood vessels are parallel to the skin surface. Angling the transducer will often disturb the flow, and new techniques for finding transverse velocities are needed. The various approaches for determining transverse velocities will be explained. This includes techniques using two-dimensional correlation (speckle tracking), multiple beams, and the new transverse modulation technique. The different advantages and disadvantages of the approaches are explained. PMID- 10829689 TI - Flow estimation using an intravascular imaging catheter. AB - Coronary flow assessment can be useful for determining the hemodynamic severity of a stenosis and to evaluate the outcome of interventional therapy. We developed a method for measuring the transverse flow through the imaging plane of an intravascular ultrasound (IVUS) catheter. This possibility has raised great clinical interest since it permits simultaneous assessment of vessel geometry and function with the same device. Furthermore, it should give more accurate information than combination devices because lumen diameter and velocity are determined at the same location. Flow velocity is estimated based on decorrelation estimation from sequences of radiofrequency (RF) traces acquired at nearly the same position. Signal gating yields a local estimate of the velocity. Integrating the local velocity over the lumen gives the quantitative flow. This principle has been calibrated and tested through computer modeling, in vitro experiments using a flow phantom and in vivo experiments in a porcine animal model, and validated against a Doppler element containing guide wire (Flowire) in humans. Originally the method was developed and tested for a rotating single element device. Currently the method is being developed for an array system. The great advantage of an array over the single element approach would be that the transducer has no intrinsic motion. This intrinsic motion sets a minimal threshold in the detectable velocity components. Although the principle is the same, the method needs some adaptation through the inherent different beamforming of the transducer. In this paper various aspects of the development of IVUS flow are reviewed. PMID- 10829690 TI - Speckle tracking for multi-dimensional flow estimation. AB - Speckle tracking methods overcome the major limitations of current Doppler methods for flow imaging and quantification: angle dependence and aliasing. In this paper, we review the development of speckle tracking, with particular attention to the advantages and limitations of two-dimensional algorithms that use a single transducer aperture. Ensemble tracking, a recent speckle tracking method based upon parallel receive processing, is described. Experimental results with ensemble tracking indicate the ability to measure laminar flow in a phantom at a beam-vessel angle of 60 degrees, which had not been possible with previous 2D speckle tracking methods. Finally, important areas for future research in speckle tracking are briefly summarized. PMID- 10829691 TI - Statistical evaluation of clutter filters in color flow imaging. AB - The filter used to separate blood signals from the tissue clutter signal is an important part of a color flow system. In this paper, statistical detection theory is used to evaluate the quality of the most commonly used clutter filters. The probability of falsely classifying a sample volume as containing blood is kept below a specified threshold. With this constraint, the probability of correctly detecting blood is calculated for all the filters. Using a measured clutter signal, we found that polynomial regression filters and projection initialized IIR filters are best among the commonly used filters. The probability of correctly detecting blood with velocity 10.1 cm/s was 0.32 for both these filters. The corresponding value for the optimal detector was 0.81, whereas a regression filter that depends on the clutter signal statistics achieved a blood detection probability of 0.72. PMID- 10829692 TI - Real-time velocimetry for evaluation of change in thickness of arterial wall. AB - We previously developed a new method, namely, the phased tracking method, for accurately tracking the movement of the heart wall and arterial wall based on both the phase and magnitude of the demodulated signals to determine the instantaneous position of an object. By this method, the local change in wall thickness during one heartbeat can be determined. We have now developed a real time system for measuring change in thickness of the myocardium and arterial wall. In this system, four high-speed digital signal processing (DSP) chips are employed for obtaining the initially developed method in real time. The tracking results for both sides of the wall are superimposed on the M (motion)-mode image in the workstation, and the thickness changes of the arterial wall are displayed in real time. Using this system, reported herein, velocity signals of the arterial wall with amplitudes less than several micrometers can be successfully detected in real time with sufficient reproducibility. The elasticity of the arterial wall is evaluated by referring to the blood pressure. In in vivo experiments, the rapid response of the change in wall thickness of the carotid artery to the dose of nitroglycerine (NTG) is evaluated for a young healthy subject and a young smoker. This new real-time system offers potential for quantitative diagnosis of early-stage atherosclerosis by the transient evaluation of the rapid response of the cardiovascular system to physiological stress. PMID- 10829693 TI - Vascular tissue characterisation with IVUS elastography. AB - Knowledge about the mechanical properties of the vessel wall and plaque is important for guiding intravascular interventional procedures and detection of plaque vulnerability. Rupture of atherosclerotic plaques is associated with acute myocardial infarction and unstable angina pectoris. In a plaque with a lipid core, the stress due to the arterial pulsation will be concentrated in the cap and a thin cap may be unable to bear this stress. In this study, the potential of intravascular elastography to characterise fibrous, fibro-fatty and fatty tissue based on their mechanical properties was investigated. Using a custom-made set up, intravascular echograms and elastograms of excised human femoral arteries were determined. High frequency r.f. data (30 MHz) were acquired using an intravascular catheter. The tissue was compressed using intravascular pressures of 80 and 100 mmHg. The cross-sections of interest were marked with a needle for matching with histology. Using cross-correlation estimation of gated echosignals, elastograms (images of the local strain) were determined. After the intravascular experiments, the specimens were fixed in formaldehyde and processed for paraffin embedding. Sections were stained with picrosirius red and alpha-actin to counterstain collagen and smooth muscle cells (SMC), respectively. Results of vessel cross-sections with fibrous and fatty plaque regions will be presented. The elastograms of these specimens show that the strain in fatty tissue is higher than the strain in fibrous material. In conclusion, these in vitro experiments on human femoral arteries indicate the potential of intravascular elastography to characterise different plaque components. PMID- 10829694 TI - High-frequency ultrasound characterization of microporous biointegrable polymers in cornea using acoustic parameters. AB - The current work deals with the use of high-frequency quantitative backscatter acoustic microscopy for the evaluation of the biointegration of microporous polymer implants used as support of artificial cornea. A three-dimensional (3-D) 80 MHz ultrasound microscope (25 microns axial resolution at focus) was used for the imaging and characterization of the progressive biointegration of polymers implanted in rabbit cornea. In-vitro and in-vivo studies were performed. Quantitative assessment of the structural changes in the biomaterial was performed using the spectral analysis of the radio frequency signal and the estimation of acoustic parameters in the 10-65 MHz frequency bandwidth. Correlation of in-vitro ultrasound data with histologic findings has shown that attenuation and backscatter coefficients are sensitive to the changes in the polymer pore content with time. Our results obtained in vivo demonstrated that 3 D 80 MHz echography coupled with quantitative characterization provide a unique tool for the non-invasive and objective follow-up of the implant biointegration and should contribute to clinical management of artificial cornea. PMID- 10829695 TI - Visualization of human umbilical vein endothelial cells by acoustic microscopy. AB - The morphology and acoustic properties of the human umbilical vein endothelial cells (HUVECs) were evaluated using a scanning acoustic microscope system. HUVECs were cultured for 4 days and exposed to the endotoxin for 4 h. The frequency of the scanning acoustic microscope was variable between 100 and 210 MHz. By changing the measuring frequency, ultrasonic amplitude and phase were measured and the quantitative value of attenuation was calculated. Before and after endotoxin stimuli, HUVECs were observed by scanning acoustic microscopy and the attenuation was measured. The acoustic images were successfully obtained to identify the outer shape of the HUVEC and the location of the nucleus in the cell. The attenuation of the nucleus is higher than that of the cytoplasm. The attenuation of the cytoplasm was increased and became inhomogeneous after endotoxin exposure. This finding would be related to the change of F-actin filaments, which is the main component of the cytoskeleton. Scanning acoustic microscopy is useful for assessing the cellular viscoelastic properties since it can detect both the morphological and acoustic changes without contacting the cellular surface. PMID- 10829696 TI - Shear strain estimation and lesion mobility assessment in elastography. AB - Elastography typically measures and images the normal strain component along the insonification/compression axis, i.e., in the axial direction. We have recently shown that, by using interpolation and cross-correlation methods of transversely displaced RF echo segments, it is possible to measure and image displacement and strain transversely to the beam with good precision. This enables the estimation and imaging of all three principal normal strain components. Generally, motion in a direction other than that in which strain is estimated may result in decorrelation noise, severely corrupting the estimates. Therefore, a correction method is applied to correct the displacement and strain estimates for decorrelating motion. In this paper, we show how corrected displacement estimates can also be used to estimate and image the shear strain components. This may allow us to identify regions of decorrelation noise in the normal strain measurement that are due to shear strain. Shear strain estimates provide supplementary information, which can characterize different tissue elements based on their mobility. In the case of breast lesions, low mobility is related to malignancy. Following an in vivo case, we show with 2D simulations how assessment of tumor mobility can be achieved with shear strain estimation. PMID- 10829697 TI - Onset of pulsatile waves in the heart walls at end-systole. AB - We have previously developed a novel ultrasonic method, namely, the phased tracking method, for accurately tracking the movement of the heart wall based on both the phase and magnitude of the demodulated signals to determine the instantaneous position of an object. With this method, it is possible to accurately detect small-amplitude velocity signals of less than a few micrometers of the heart wall that are superimposed on the motion of the heart wall due to the heart beat. There are several remarkable pulsatile waves during one cardiac cycle in the resultant velocity signals, some of them being commonly obtained for both healthy subjects and patients. These pulsatile waves cannot be recognized in standard echocardiography M-mode images. In this paper, by focusing on one pulsatile wave that occurs around the end-systole, the physiological meaning of these is considered based on various in-vivo experiments. The pulsatile wave measured by this novel ultrasonic method will offer potential for a quantitative assessment of myocardial viability. PMID- 10829698 TI - Spectral estimators in elastography. AB - Like velocity, strain induces a time delay and a time scaling to the received signal. Elastography typically uses time delay techniques to indirectly (i.e. via the displacement estimate) measure tissue strain induced by an applied compression, and considers time scaling as a source of distortion. More recently, we have shown that the time scaling factor can also be spectrally estimated and used as a direct measure of strain. Strain causes a Doppler-like frequency shift and a change in bandwidth of the bandpass power spectrum of the echo signal. Two frequency shift strain estimators are described that have been proven to be more robust but less precise when compared to time delay estimators, both in simulations and experiments. The increased robustness is due to the insensitivity of the spectral techniques to phase decorrelation noise. In this paper we discuss and compare the theoretical and experimental findings obtained with traditional time delay estimators and with the newly proposed spectral methods. PMID- 10829699 TI - Angle matching in intravascular elastography. AB - Intravascular elastography is a new technique to obtain mechanical properties of the vessel wall and plaque. Mechanical information of vascular tissue is important for characterisation of different plaque components, detection of plaque vulnerability and thus choosing the proper interventional technique. The feasibility of the technique is investigated using phantoms and diseased human arteries. These studies demonstrated that elastography reveals information that is unavailable or inconclusive from the echogram alone. The technique is based on the principle that tissue strain is directly related to its mechanical properties. In intravascular elastography, the tissue is compressed using different intravascular pressures. The strain is determined using cross correlation techniques of the radio frequency (r.f.) signals. Reliable strain estimates are only obtained when signals of corresponding tissue are correlated. Owing to catheter motion, off-centre position and non-uniform rotation of the intravascular transducer, the r.f. traces at low and at high pressure may be misaligned. Four algorithms are tested to track the corresponding ultrasound signals. Three methods (l1norm, l2norm and cross-correlation) are applied on the r.f. signal and one (l1norm) on the envelope (speckle tracking). Simulations are performed to obtain a data set with a priori knowledge of the scattering particles positions in the tissue at high and low pressure. Different positions of the catheter in the lumen, compression levels of the material and signal-to noise ratios (SNRs) are investigated. Finally, these findings are corroborated with a phantom experiment in a water tank. From the simulations, it can be concluded that the speckle tracking algorithm has the best performance, under all circumstances. The performance decreases with larger eccentricity of the catheter and larger compression of the material. The SNR is only of minor influence. The speckle tracking algorithm has also the best performance in the phantom experiment. The performance of the speckle tracking algorithm is better than the three r.f.-based algorithms. For intravascular elastography, implementation of this method may improve the quality of the elastogram. PMID- 10829700 TI - Ultrasound transmission tomography image distortions caused by the refraction effect AB - This paper presents the influence of ultrasonic beam ray refraction effect on the internal object structure imaging using the ultrasound transmission tomography method (UTT), in parallel ray projection geometry, for ultrasonic pulse run-time measurements. Simulation calculations carried out using specially elaborated software were verified experimentally by means of real measurements on a research set-up for UTT. Simple ways to minimize image distortions caused by the refraction effect were also suggested. PMID- 10829701 TI - Quantitative determination of contact stiffness using atomic force acoustic microscopy AB - Atomic force acoustic microscopy is a near-field technique which combines the ability of ultrasonics to image elastic properties with the high lateral resolution of scanning probe microscopes. We present a technique to measure the contact stiffness and the Young's modulus of sample surfaces quantitatively, with a resolution of approximately 20 nm, exploiting the contact resonance frequencies of standard cantilevers used in atomic force microscopy. The Young's modulus of nanocrystalline ferrite films has been measured as a function of oxidation temperature. Furthermore, images showing the domain structure of piezoelectric lead zirconate titanate ceramics have been taken. PMID- 10829702 TI - Texture measurement of shaped material by impulse acoustic microscopy AB - All the microstructural parameters involved in metallurgical processes are difficult to determine directly on a shaped material. The aim of this paper is to use an impulse line-focus acoustic microscope (LFAM) as a non-destructive alternative to X-ray diffraction for measuring texture of slightly anisotropic materials. We apply it to characterize the rolling and annealing texture for tantalum sheets. PMID- 10829703 TI - Space- and time-resolved Brillouin light scattering from nonlinear spin-wave packets AB - We have constructed a new Brillouin light scattering apparatus, based on the Sandercock multipass tandem interferometer design, for space- and time-resolved investigations of nonlinear wave packets in thin films. We have applied the method to studies of nonlinear spin-wave pulse propagation in yttrium iron garnet (YIG) films. Spatial resolution is achieved by scanning the laser spot across the YIG film surface, and temporal resolution is obtained by measuring the elapsed time between the launch of spin-wave pulses by an applied microwave pulse and the arrival of the respective inelastically scattered photons at the detector. We report the observation of nonlinear self-focusing of wave beams and pulses in one and two dimensions, the formation of one-dimensional envelope solitons, and of strongly localized, two-dimensional wave packets, 'spin-wave bullets', analogous to 'light bullets' predicted in nonlinear optics. By generating two counter propagating wave pulses, pulse collision experiments were performed. We show that quasi-one-dimensional envelope solitons formed in narrow film stripes ('waveguides') retain their shapes after collision, while two-dimensional spin wave packets formed in wide YIG films are destroyed in collision. PMID- 10829704 TI - Surface Brillouin scattering of opaque solids and thin supported films AB - Surface Brillouin scattering (SBS) has been used successfully for the study of acoustic excitations in opaque solids and thin supported films, at both ambient and high temperatures. A number of different systems have been investigated recently by SBS including crystalline silicon, amorphous silicon layers produced by ion bombardment and their high temperature recrystallisation, vanadium carbides, and a nickel-based superalloy. The most recent development includes the measurement of a supported gold film at high pressure. The extraction of the elastic constants is successfully accomplished by a combination of the angular dependence of surface wave velocities and the longitudinal wave threshold within the Lamb shoulder. The application of surface Green's function methods successfully reproduces the experimental SBS spectra. The discrepancies often observed between surface wave velocities and by ultrasonics measurements have been investigated and a detailed correction procedure for the SBS measurements has been developed. PMID- 10829705 TI - Elastic measurements of layered nanocomposite materials by Brillouin spectroscopy AB - Surface Brillouin spectroscopy makes it possible to measure surface elastic wave propagation parameters at frequencies up to 20 GHz or more. This enables us to measure the elastic properties of surface layers only a small fraction of a micrometre thick. The wavelength and incident angle of the light determine the wavenumber of surface elastic waves (SAW) that scatter the light inelastically, and their frequency can be found by measuring the change in wavelength of the scattered light. By analysing the elastic wave modes present in the surface, the elastic properties can be deduced. We have used this technique to measure the elastic properties of layered nanocomposite materials, which are widely used in the packaging industry. 12 microns polymer films (PET) were coated with glass oxide layers of thickness as little as 25 nm, to give transparent nanocomposite structures with excellent gas barrier properties. In order to understand and model the behaviour of these films under deformation, it is necessary to determine the elastic properties of the different layers. Evaluation of the elastic properties presents several challenges. First, the oxide layers are much thinner than the wavelengths of the surface phonons in surface Brillouin spectroscopy (and hence the depth probed), which usually lie in the range 250-500 nm. The anisotropic elastic properties of the PET substrate must therefore be measured accurately, and this can be done using bulk Brillouin spectroscopy. Second, a thin layer of metal (usually 10-20 nm) must be deposited on the glass surface so that the surface phonons scatter the light effectively. The elastic properties of the glass layer can then be deduced from surface Brillouin spectroscopy measurements, by simulating the surface wave modes of the metal/glass/polymer composite, and adjusting the parameters to give the best fit. In this way it is possible to observe how the properties of the glass vary as a function of thickness, and in turn to understand how to improve systematically the properties under deformation. PMID- 10829706 TI - Brillouin scattering study of epoxy adhesive layers during cure AB - The isothermal curing process of thin epoxy adhesive layers (mixture of a bisphenol A-based epoxy prepolymer and an aliphatic diamine curing agent) has been investigated. The Brillouin scattering technique with 90 degrees A scattering geometry enables simultaneous measurements of longitudinal and shear wave properties in the GHz range. Observed longitudinal and shear wave velocities showed similar changes during cure. They rapidly increased and gradually became constant, reflecting the elastic changes of the epoxy layer. The final velocities, however, clearly depended on the curing temperature. Taking the glass transition process of epoxy resins into consideration, these curing behaviors in thin layers are discussed. PMID- 10829707 TI - Laser picosecond acoustics in isotropic and anisotropic materials AB - We present experimental results concerning the laser generation of picosecond acoustic pulses and their propagation in isotropic and anisotropic materials. We make use of a conventional reflectance detection technique as well as interferometric detection to probe the real and imaginary changes in reflectance. We also demonstrate the detection of transverse acoustic waves by mode conversion at an interface between an isotropic polycrystalline film and an anisotropic substrate. PMID- 10829708 TI - On the rise time of ultrafast surface displacement of laser-irradiated surface AB - It is demonstrated that acoustic attenuation does not influence the rise time of transient laser-induced displacement of a surface. PMID- 10829709 TI - Quantitative estimation of ultrasonic attenuation in a solid in the immersion case with correction of diffraction effects AB - This paper presents a method of diffraction correction for the log-spectral difference method to estimate quantitatively attenuation of a solid in the immersion case. The correction method is established based on the angular spectrum approach that is used to calculate the echoes from the front and back surfaces of the immersed solid. An example is given of a copper plate submerged in water and inspected by a linear array with a cylindrically curved surface. The correction method is first applied to a theoretical estimation of attenuation which is linearly dependent on frequency. The results have shown that the evaluated attenuation coefficient is in excellent agreement with the exact value. Then the method is applied to a real situation, in which the results have shown that the method yields reasonable evaluated attenuation values. This work has demonstrated that the method is able to correct effectively the diffraction effect so as to achieve a quantitative estimation of attenuation. PMID- 10829710 TI - Ultrasonic monitoring of film condensation for applications in reduced gravity AB - The film condensation of vapor on a cooled, downward-facing surface is a complex dynamic fluid phenomenon. This paper will describe the theoretical basis, instrumentation approach, and experimental results for a method for ultrasonic monitoring of film condensation. The ultrasound measurement system utilizes a 10 MHz broadband contact transducer, operating in pulse-echo mode. By appropriate signal processing, condensation layers over a wide thickness range, from much less than a wavelength to several wavelengths, can readily be measured. PMID- 10829711 TI - Temperature dependence of diffuse field phase AB - Diffuse fields, which have scattered from microstructure or reflected from walls so much as to prohibit conventional analyses, are usually examined by means of the time evolution of their ultrasonic spectral energy density. The phase information is usually discarded as resisting analysis. The phase, while unpredictable is, however, robust; according to theory it remains constant if source and receiver are not disturbed. Nevertheless, in practice we do observe slow drifts of phase over time scales of minutes. Here we examine the hypothesis that the phase drifts are due to temperature fluctuations. Temperature changes on cooling from 40 degrees C to room temperature were monitored and compared with changes in diffuse field phase. It was found that the reverberant ultrasonic field in a 7 cm aluminum block evolves with temperature in a manner that is in accord with published data on the temperature dependence of the ultrasonic velocities. Our 1 MHz transient source gives rise to a complex waveform that is observed to undergo an almost pure dilation. The precision with which this shift can be measured approaches 20 ns. This is remarkable when compared with the 100 ms travel time of the signal. Thus the temperature dependence of elastic wave speed is measured with a precision limited by the precision of one's thermometer. The signal is also found to suffer some distortion which, it is suggested, is related to the different rates of change of longitudinal and shear speeds. The corresponding prediction for the degree of distortion is found to be in accord with measurements. PMID- 10829712 TI - Pencil method in elastodynamics: application to ultrasonic field computation AB - The principles of pencil elastodynamics and, in more detail, some selected applications of pencil techniques to elastodynamics are described. It is shown how a systematic use of a matrix representation for the wave front curvature and for its transformations simplifies the handling of arbitrary pencils and, consequently, the field computations. Pencil matrix representations for the propagation into homogeneous solids made of isotropic or anisotropic media are derived. The use of matrix representations for pencil reflections on, or refractions through, arbitrarily curved interfaces, together with matrix representations for propagation into homogeneous media, allow us to derive an overall matrix formulation for elastodynamic propagation into complex heterogeneous structures. Combined with the classical Rayleigh integral to account for transducer diffraction effects, the proposed theory is applied to the prediction of ultrasonic fields radiated into complex structures by arbitrary transducers. Examples of interest for application to ultrasonic non-destructive testing are given. PMID- 10829713 TI - Signal-to-noise ratio enhancement based on the whitening transformation of colored structural noise AB - In the ultrasonic testing and evaluation of highly scattering materials (i.e. non homogeneous media such as composites, layered and clad materials) structural noise is an important limitation to the visibility of flaw echoes. This noise cannot be reduced by conventional linear filtering or by time-averaging techniques. In order to enhance the defect-to-background noise ratio (SNR), many different algorithms have been developed over the years. This work analyzes three new strategies for SNR enhancement based on the whitening transformation of the colored structural noise. By using this transformation, the small spectral differences between noise and flaw echoes are exploited, thereby allowing an improvement in the visibility of the flaw. PMID- 10829714 TI - Elaboration of some signal processing algorithms in ultrasonic techniques: application to materials NDT AB - In ultrasonic techniques, information on defect characterization possibilities has required more evolved technique development than classical methods. To obtain a high probability of defect detection, these methods use signal-processing algorithms in order to enhance the signal-to-noise ratio. These methods are also used to discriminate between planar and volumetric defects. In this paper, some signal-processing algorithms are developed and implemented on a computer to allow their utilization in real-time processing of ultrasonics NDT results. PMID- 10829715 TI - Laser ultrasonic surface wave inspection of alumina ceramics of varying density AB - In this paper, the surface acoustic wave velocity results acquired from the inspection of specially manufactured and characterised alumina ceramic materials are presented. Ultrasonic velocity data of alumina-based ceramics in the range 60 100% theoretical density was generated utilising non-contacting laser-ultrasonic measurements based on laser generation and detection of surface acoustic waves with the objective of creating a routine technique for industrial advanced alumina inspection. With linear fitting the surface acoustic wave velocity data serves as a calibration graph for using laser ultrasonics for routine monitoring of alumina. A second laser ultrasonic technique based on the laser generation and foil transducer detection of surface acoustic waves was used to validate the surface acoustic wave velocities measured by the laser generation/detection technique. PMID- 10829716 TI - Ultrasonic measurement of the diffusion bond strength AB - This paper presents an ultrasonic method for measurement of diffusion bond strength between two identical materials. The method requires a single normal incidence ultrasonic measurement. When a diffusion bond is not perfect some ultrasonic energy is reflected from the interface separating the two substrates. Amplitude of a reflected signal correlates qualitatively with the bond strength. However, a quantitative correlation is difficult to establish since reflected signal amplitude depends on parameters of the test equipment. We introduce a spectroscopic analysis of a reflected signal to measure the bond strength. We show experimentally that an imperfect diffusion bond can be characterized by a single parameter: interfacial spring stiffness. The stiffness can be determined from the reflected signal spectrum and does not depend on testing equipment. It is also demonstrated that for a particular diffusion bonding procedure as applied to tool steel the interfacial stiffness correlates quantitatively with bond strength. PMID- 10829717 TI - Measurement of the dispersion relation of guided non-axisymmetric waves in filament-wound cylindrical structures AB - To determine the dispersion relation, guided waves are excited in specimens over a broad frequency range. The surface displacements are measured over time and space. The recorded data are analysed using a quasi-three-dimensional spectrum estimation algorithm. In the time domain a fast Fourier transform is used to extract the frequencies. To obtain the wave numbers, in space a two-dimensional matrix-pencil approach is applied to the data set. Using a suitable constitutive model (transversely isotropic or orthotropic) dispersion curves are calculated. Good agreement was found between the experimental and the numerically calculated dispersion relations after adjusting the material parameters. Since the dispersion relation of a structure depends on the mechanical material properties frequency-dependent material parameters can be extracted from the above-mentioned relation between frequency and wave number. PMID- 10829718 TI - Modeling of integrated Lamb waves generation systems using a coupled finite element-normal modes expansion method AB - As part of the research work on Smart Materials and Structures, the development of self-monitoring materials is an emerging issue. In the case of plate-shaped structures, Lamb waves can be used for their relevant properties: long-range propagation, sensitivity to internal flaws and whole-thickness interrogation. This concept requires the use of thin piezoelectric transducers integrated to the structure. Since it is of primary importance to be able to control the generated modes, a suitable modeling technique of this kind of system has been tested on different cases of practical interest. The model uses a coupled finite element normal modes expansion approach, which allows one to consider either the case of bonded or embedded transducers. The results presented deal with examples of multi element transducers integrated to composite materials. The influence of parameters such as the dimensions, positions and relative excitation delays of the transducers is studied. PMID- 10829719 TI - Transient elastodynamic model for beam defect interaction: application to non destructive testing AB - Modeling tools have been developed at the French Atomic Energy Commission (CEA) for the simulation of ultrasonic non-destructive testing inspections. In this paper the model for the prediction of echoes arising from defects within a piece (Mephisto) is presented and some examples are given and compared with experimental results. The model for computing wave defect interaction is based on Kirchhoff's approximation, and uses the principle of reciprocity and a mode-by mode (between the transducer and the defect) calculation of the echoes. It accounts for possible mode conversions. These approximations and other approximations for the radiated field incident on the defect allow us to obtain a formulation of the echo received at the transducer, which is able to be computed rapidly. PMID- 10829720 TI - Porosity estimation of concrete by ultrasonic NDE AB - The increasing number of concrete structures with symptoms of premature deterioration due to environmental action demands procedures to estimate the durability of this type of component. Concrete durability is related to porosity, which determines the intensity of interactions of the material with aggressive agents. The pores and capillaries inside the structure facilitate the destructive processes that generally begin in the surface. In this work, an ultrasonic NDE technique to estimate the porosity of concrete is developed. The method is based on the analysis of the mechanical behaviour of mortar probes built with calibrated sand, in which the concentration of water-cement mixture has been varied. In this sense, data of sound velocity are correlated with data of porosity, which have been previously measured by destructive measurements. PMID- 10829721 TI - Acoustic inspection of bond strength of steel-reinforced mortar after exposure to elevated temperatures AB - In order to evaluate the bond strength between the reinforcement and concrete after fire damage, a combination of acoustic through-transmission and pull-out tests were used. Previous studies have shown a 25% decrease in the ultrasonic pulse velocity at 90% of the maximum load at room temperature. The specimens were kept in the oven at an elevated temperature for 1, 2, or 3 h. They were then removed and cooled to room temperature. Inspection was conducted using a high power ultrasonic pulse velocity system while a pull-out load was applied. The correlation between preheated temperature, acoustic wave velocity, and the applied load was analyzed. Initial results show that bond strength and pulse velocity decreased substantially as the temperature or the heating time increased. PMID- 10829722 TI - Modeling of propagation and echo formation in a multilayered structure AB - In the aim of simulating the ultrasonic inspection of multilayered structure, we propose a hybrid model, based on transfer matrices and ray tracing formalisms. This approach allows one to predict the response of structures containing defects of finite size such as delaminations or adhesion defect. PMID- 10829723 TI - Multiplexing and transmission of RF signals using an optical fiber AB - Ultrasonic non-destructive testing systems designed to control huge structures normally use several transducers in the reception stage. To avoid increasing the cost of electronics, a multiplexer is used to send all received signals to the same processing module. Traditionally, transmission of such signals is carried out using copper cables. For special applications (i.e. continuous monitoring of nuclear plants) metallic cables are not suitable because of their high sensitivity to electromagnetic perturbations. Moreover, the multiplexing is made electronically. When the distance between the transducers and the reception unit is large and/or electromagnetic noise is important, signal degradation takes place. The proposed system implements the transmission and multiplexing of ultrasonic electrical signals obtained by means of broadband transducers (up to 1 MHz), using an optical fiber. Optical fibers are made of dielectric materials (silica or plastic) so they are inherently passive to electromagnetic noise. Wavelength division multiplexing is utilized for adding channels to the system by means of fiber optic couplers and different light sources. The wavelengths of the optical signals utilized are located far apart in the optical spectrum in order to avoid serious crosstalk in transmission. The limit to the number of multiplexed channels depends on the optical fiber selected, the spectrum of the light sources and the wavelength division multiplexers or couplers utilized. PMID- 10829724 TI - Water content and its effect on ultrasound propagation in concrete--the possibility of NDE AB - It is known that water content or moisture affects the strength of concrete. The purpose of this study is to examine the possibility of the NDE of concrete from a knowledge of the relationship between water content and ultrasonic propagation in concrete. The results of measurements made on the ultrasound velocity and the frequency component on ultrasonic propagation as a function of the water content in concrete are reported. Test pieces of concrete made from common materials were made for the fundamental studies. The test piece dimensions were 10 cm in diameter and 20 cm in length. Test pieces were immersed in water for about 50 days to saturate them. To measure the effect of different water contents, test pieces were put in a drying chamber to change the amount of water between measurements. This procedure was repeated until the concrete was completely dried and the weight no longer changed. Water contents were defined as weight percentage to full dried state. Thus water content could be changed from 8% to 0%. Using the pulse transmission method, ultrasonic propagation in the frequency range 20 to 100 kHz was measured as a function of water content. The sound velocity varied gradually from 3000 m/s to 4500 m/s according to the water content. The frequency of maximum transmission also depended on the water content in this frequency range. It is considered that the ultrasonic NDE of concrete strength is feasible. PMID- 10829725 TI - Dependence of pulser driving responses on electrical and motional characteristics of NDE ultrasonic probes AB - Acoustic performance in ultrasonic transmitters can be improved by means of a suitable electrical driving response and matching/tuning networks. It is important to predict this electrical response, but doing so is not easy because it departs notably from the nominal pattern with the loading probes. In practice, the analysis of HV pulser spikes in NDE applications requires fairly complex models in the transient regime and, in addition, non-linear problems could arise, especially in the case of tuned transmitters. In this paper, the most relevant influences of loading characteristics of NDT ultrasonic probes on the pulser electrical driving responses are evaluated in time and frequency domains. Conventional pulse generators and typical NDE pulsers are considered. Driving responses are analysed across commercial ultrasonic probes and, alternatively, across similar purely electrical loads. Distinct influences on pulser responses from electrical and motional sections of the probes are identified. All these aspects are studied on the basis of experimental and computer results. PMID- 10829726 TI - Single-bubble sonoluminescence in microgravity AB - Single-bubble sonoluminescence refers to the emission of light from an acoustically trapped bubble undergoing highly nonlinear, presumably radial oscillations. The intensity of the emitted light depends strongly on the forcing pressure, and is limited by the development of instabilities that ultimately results in the extinction of the bubble. In this article, we discuss a possible contributing factor for the generation of instabilities; specifically, we examine the effect of the gravitational force on a sonoluminescence bubble. PMID- 10829727 TI - Dependence of single-bubble sonoluminescence on ambient pressure AB - Kondic et al.'s theory makes several specific predictions on the dependence of single-bubble sonoluminescence (SBSL) on ambient pressure. We have carried out experiments to verify these predictions for air bubbles in a water-glycerine mixture at about 17.5 kHz. The results show an increase in SBSL with reduced ambient pressure down to a critical value below which SBSL is extinguished. The results are all in good agreement with Kondic et al.'s theory and are also compatible with the dissociation hypothesis of Lohse et al. PMID- 10829728 TI - Leaky wave detection at air-solid interfaces by laser interferometry AB - A recently developed optical heterodyne interferometer is proved to be sensitive to detect ultrasonic waves leaking out from metals (and from non-metals as well) several centimeters away from the metal surface in air. This measurement is based on the detection of the optical index variation in air due to the leaky wave. Experiments were carried out using both ultrasonic leaky Rayleigh and leaky Lamb waves in the low-megahertz region. Optical measurements of these leaky wave velocities in semi-infinite materials and plates, which are related to elastic constants of the materials, showed a good correlation to the predicted values. Optical measurements of the amplitude of the leaky waves were used to obtain attenuation coefficients, which correlate to the structural integrity of the materials. Surface and near-surface defects were also detected. This optical method can be used for metals with low-reflectivity surfaces. Thus, it can be used for non-polished surfaces and can be extended for non-contact, non destructive evaluation applications. PMID- 10829729 TI - On the polarization of light diffracted by ultrasound AB - If light is diffracted by ultrasound in an isotropic medium with acoustically induced birefringence, the state of polarization is modified in each order of diffraction with respect to the initial state of polarization of the incident light wave. In the present paper, some polarization effects are discussed in the case of normal light incidence. In general a rotation of the main polarization plane occurs, together with a change of the ellipticity. However, while the former effect always takes place, the latter only occurs in the case of ultrasonic light diffraction of the intermediate type. Some experimental measurements are included in case of argon laser light being diffracted by an ultrasonic wave propagating in fused silica (SiO2). PMID- 10829730 TI - Development of an optical fiber hydrophone with fiber Bragg grating AB - A new type of optical fiber hydrophone is constructed with a fiber Bragg grating (FBG) based on the intensity modulation of laser light in an FBG under the influence of sound pressure. The FBG hydrophone shows linearity, with dynamic range about 70 dB. It can measure amplitude and phase of an acoustic field in real time, and operates in a wide range of acoustic frequency, at least from 1 kHz to 3 MHz. No signal distortion is observed in the detected signal. Because of the simplicity in its operating principle and geometry, an FBG hydrophone is expected to be an acoustic sensor of high practicality compared to a conventional optical fiber hydrophone. PMID- 10829731 TI - Collinear acousto-optic diffraction using two nearby sound frequencies AB - Collinear acousto-optic spectrometers with sound consisting of one frequency have a spectral transfer function with many good characteristics, such as a narrow bandwidth, ... the side lobes, however, are rather high. To reduce these side lobes, one can try to apodize the sound in the acousto-optic cell by using a sound wave consisting out of two frequencies. At certain times, this produces a more smoothened sound field in the cell than in the one-frequency case. The resulting transfer function will contain lower side lobes. Also, the influence of all the sound field parameters--the amplitude, frequency difference, and time--on the transmission function is studied. Out of these results, better-performing collinear spectrometers can be designed. PMID- 10829732 TI - Enhancement of photoelastic constant of optical thin film waveguide assisted by surface acoustic wave AB - In the present research, we proposed a method of controlling the photoelastic constant using surface acoustic waves, which had not previously been reported, and carried out experimental studies thereof. A Bragg diffraction was carried out to determine the photoelastic constants of Ta2O5. As a result, it is confirmed that the photoelastic constant of a Ta2O5 thin film undergoing a sputtering process, during which surface acoustic waves were excited on the substrate, was about 2.19-2.27 times larger than those of thin films on which surface acoustic waves were not excited. PMID- 10829733 TI - Prolonged acousto-optic interaction with Lamb waves in crystalline plates AB - The propagation and acousto-optic interaction of Lamb modes in an anisotropic plate of tellurium dioxide (TeO2) are studied numerically and analytically. In the case of a Y-cut X-propagating TeO2 plate, the very high elastic anisotropy of the crystal greatly modifies the dispersion curves, giving rise to their multiple oscillations. The existence ranges of backward Lamb modes increase with the mode order contrary to the case of isotropic plates. The quasi-collinear light scattering by Lamb waves is considered. Owing to the structure of Lamb wave field, a simultaneous light diffraction at two different optical frequencies can take place while Lamb waves are excited only at the single frequency. It is demonstrated with the Z-cut (110)-propagating plate that a small change in the acoustic frequency can result in a significant shift in the frequency of the scattered light. PMID- 10829734 TI - Analysis of acoustic emission signals and monitoring of machining processes AB - Monitoring of a machining process on the basis of sensor signals requires a selection of informative inputs in order to reliably characterize and model the process. In this article, a system for selection of informative characteristics from signals of multiple sensors is presented. For signal analysis, methods of spectral analysis and methods of nonlinear time series analysis are used. With the aim of modeling relationships between signal characteristics and the corresponding process state, an adaptive empirical modeler is applied. The application of the system is demonstrated by characterization of different parameters defining the states of a turning machining process, such as: chip form, tool wear, and onset of chatter vibration. The results show that, in spite of the complexity of the turning process, the state of the process can be well characterized by just a few proper characteristics extracted from a representative sensor signal. The process characterization can be further improved by joining characteristics from multiple sensors and by application of chaotic characteristics. PMID- 10829735 TI - Some aspects of AE application in tool condition monitoring AB - Acoustic emission (AE) is rather a well-known form of non-destructive testing. In the last few years the technology of the AE measurement has been expanded to cover the area of tool condition monitoring. The paper presents some experience of Warsaw University of Technology (WUT) in such applications of AE. It provides an interpretation of common AE signal distortions and possible solutions to avoid them. Furthermore, a characteristic study of several different AE and ultrasonic sensors being used in WUT is furnished. Evaluation of the applicability of some basic measures of acoustic emission for tool condition monitoring is also presented in the paper. Finally paper presents a method of the catastrophic tool failure detection in turning, which uses symptoms other than the direct magnitude AERMS signal. The method is based on the statistical analysis of the distributions of the AERMS signal. PMID- 10829736 TI - Information contained in the radiating ultrasound during ultrasonic welding AB - During ultrasonic welding, unnecessary ultrasound together with audible sound is radiated into the air. Audible sound is noisy and uncomfortable, and ultrasound may have bed effects on adjacent equipment. However, it is considered that these sounds potentially contain useful information such as welding state. This article reports the relationship between radiated ultrasound and the change of the welding state during ultrasonic welding. It is known that the welding state can be presumed from the change of the mechanical load impedance, which can be calculated from resonant frequency, motional voltage and the driving current of the vibrating system for the welder when a constant-velocity motional-feedback power oscillator is used. In this study, radiated ultrasound picked up with a microphone and an amplifier is recorded and analyzed with a personal computer. Analyzed results are compared with the change of the mechanical load impedance that is also calculated from the recorded data of the resonant frequency, motional voltage and the driving current of the vibrating system for the welding. Experiments were made on the ultrasonic welding of plastic rods. The relationship between the parameters of radiated ultrasound and the electrical parameters of the vibrating system were compared. The peak amplitude of the radiated ultrasound and the mechanical impedance versus time were very similar. It is shown in this study that radiated ultrasound contains lots of useful information. PMID- 10829737 TI - Acoustic emission monitoring of high speed grinding of silicon nitride AB - Acoustic emission (AE) monitoring of a machining process offers real-time sensory input which could provide tool condition and part quality information that is critical to effective process control. However, the choice of sensor, its placement, and how to process the data and extract useful information are challenging application-specific questions which researchers must consider. Here we report an effort to resolve these questions for the case of high speed grinding of silicon nitride using an electroplated single-layered diamond wheel. A grinding experiment was conducted at a wheel speed of 149 m s-1 and continued until the end of the useful wheel life. AE signal data were then collected for each complete pass at given grinding times throughout the useful wheel life. We found that the amplitude of the AE signal monotonically increases with wheel wear, as do grinding forces and energy. Furthermore, the signal power contained in the AE signal proportionally increases with the associated grinding power, which suggests that the AE signal could provide quantitative information of wheel wear in high-speed grinding, and could also be used to determine when the grinding wheel needs replacement. PMID- 10829738 TI - Determination of plate source, detector separation from one signal AB - We address the problem of locating a transient source, such as an acoustic emission source, in a plate. We apply time-frequency analysis to the signals detected at a receiver. These highly dispersive and complex waveforms are measured for source-receiver separations ranging from 40 to 180 plate thicknesses and at frequencies such that 10 to 20 Rayleigh-Lamb branches are included. Reassigned, smoothed, pseudo-Wigner-Ville distributions are generated that exhibit the expected sharp ridges in the time-frequency plane, lying along the predicted frequency-time-of-arrival relations. The source-receiver separation can be determined from such plots. PMID- 10829739 TI - Ultrasound conditioning of suspensions--studies of streaming influence on particle aggregation on a lab- and pilot-plant scale AB - Suspension conditioning by ultrasound induced particle aggregation in a 3.3 MHz standing wave field was investigated in lab- and pilot scale sedimentation reactors (sonicated volume approx. 375 ml and 10 l). These were equipped with a variable number of acoustically transparent film (ATF) elements, inserted perpendicular to the sound propagation direction. ATFs control the development of undesired large-range Eckart-type acoustic streaming and thermal convection. ATF insertion led to increased separation efficiency especially for particles smaller than 10 microns. Results with a varied geometric set-up indicate the existence of an optimum for the number of ATFs inserted. Optical investigation of particle behaviour in different pilot plant configurations showed reduced streaming within the divided 24 mm pathlength sonic chamber, whereas at 100 mm pathlength, turbulent streaming dominated. PMID- 10829740 TI - Viability of plant cell suspensions exposed to homogeneous ultrasonic fields of different energy density and wave type. AB - Exposure of Petunia hybrida cell suspensions to ultrasound at a frequency of 2.43 MHz in a standing wave field at an energy density of 70 Jm-3 (pressure amplitude of 0.78 MPa) decreased their mean viability to 35% after 20 min of sonication. A comparison of propagating wave and standing wave treatments at equal frequency (2.15 MHz) and energy density (8.5 Jm-3) showed, in the first case, a rapid decline in mean viability of cells (to 30% after 10 min of sonication) and, in the second case, a retaining of the initial viability (95%), respectively. Cells sonicated 4 days after subculture were more sensitive than cells sonicated 2 or 6 days after transfer to new culture medium. It was concluded that cellular viability depends primarily on the acoustic energy density, the exposure time, and the mechanical properties of the cells determined by age. As a consequence of the trapping of cells in the anti-node planes of the standing wave, propagating wave fields reduced cellular viability compared with standing wave fields at equal energy density. PMID- 10829741 TI - Viability of yeast cells in well controlled propagating and standing ultrasonic plane waves. AB - Recent studies have shown that there is no loss of cell viability when the cells are subjected to ultrasonic standing wave fields in acoustic cell retention systems. These systems are characterised by waves that spatially vary in pressure amplitude in the direction of sound propagation. In this work an anechoic 'one dimensional' sonication chamber has been developed that produces propagating waves, which differ from standing waves in that the pressure amplitude remains constant as the wave travels in a medium with negligible attenuation. The viability of yeast cell suspensions as a function of treatment time was investigated during exposure to both standing and propagating wave fields with frequencies slightly above 2 MHz. The influence of 12% (vol/vol) of ethanol in water on the spatial arrangement of the cells in suspension was also studied. Changes in yeast cell morphology caused by the different types of suspension media and the ultrasonic treatment were examined by transmission electron microscopy (TEM). The agglomeration of yeast cells within the pressure nodal planes appears to minimise damaging effects due to ultrasonic fields. PMID- 10829742 TI - Analytical scale ultrasonic standing wave manipulation of cells and microparticles. AB - The ultrasonic standing-wave manipulation of suspended eukaryotic cells, bacteria and submicron latex or silica particles has been examined here. The different systems, involving plane or tubular ultrasonic transducers and a range of acoustic pathlengths, have been designed to treat suspension volumes of analytical scale i.e. 5 ml to 50 microliters for both sample batch and 'on-line' situations. Frequencies range from 1 to 12 MHz. The influence of secondary cell cell interaction forces in determining the cell concentration dependence of harvesting efficiency in batch sedimentation systems is considered. Applications of standing wave radiation forces to (1) clarify cell suspensions, (2) enhance particle agglutination immunoassay detection of cells or cellular products and (3) examine and enhance cell-cell interactions in suspension are described. PMID- 10829743 TI - Application of high-power ultrasound to enhance fluid/solid particle separation processes AB - The separation of fine particles from gases or liquids is a topic of permanent industrial attention. The use of ultrasonic energy to assist conventional separation techniques seems to be very promising. The adequate applications of high-intensity ultrasonic fields may contribute to improve the efficiency and capacity of the separation methods presently used. The specific mechanisms to ultrasonically enhance separation processes basically depend on the medium to be treated. In gas suspensions, where very fine particles have to be removed, ultrasonic action involves agglomeration of particles in order to increase their size and, consequently, to improve collection efficiency of conventional filters. In liquid suspensions, agglomeration is, in general, less efficient than in gases. Nevertheless, the ultrasonic energy is useful to dewater fine-particle high-concentration suspensions such as slurries and sludges. This paper deals with the application of acoustic energy to assist fluid/solid separation processes in gas and liquid suspensions and presents some theoretical and experimental results in specific applications. PMID- 10829744 TI - Novel wedge-shaped doubly inclined chamber for the flow-through separation of suspended particles AB - Combined standing and propagating wave modes have previously been successfully used by the authors for simultaneous agglomeration and transportation of small particles suspended in still water. The present study of this method with flowing water, using a 120 x 350 x 13 mm3 agglomerator, confirmed that the proposed method is applicable with suspended SiO2 particles of varying size. The process was found to be most efficient at flow rates below a certain threshold, which varies with particle size. This threshold was found to be 5 ml/s for a particle size of 7.9 microns at an ultrasonic frequency of 2 MHz corresponding to 0.74 mm wavelength. The existence of a maximum particle transport velocity was demonstrated. PMID- 10829745 TI - The use of ultrasonic standing waves to enhance optical particle sizing equipment AB - Fluid dynamics modelling augmented with routines to simulate acoustic forces on aerosol particles has been used to investigate the potential of combining ultrasonic standing wave fields with optical particle analysis equipment. Simulations of particle dynamics in airstreams incorporating acoustic forces predict that particles in the 1-10 microns diameter range may be effectively focused to the velocity nodes of the standing wave field. Particles move to the velocity nodes within tens of milliseconds for acoustic frequencies of 10-100 kHz and at an acoustic energy density of 100 Jm-3. Larger particles are predicted to move to the velocity antinodes within similar times; however, there is a crossover region at approximately 15-20 microns particle diameter where longer times are predicted due to the competing forces driving particles to the vibration node and antinode. With sufficient transverse flow velocities the models predict that disturbances due to acoustic streaming can be overcome and a useful degree of focusing achieved for the aerosol particles. Results from a model demonstrating sampling and acoustic focusing of 3-9 microns aerosol particles to a 200 microns wide analysis area are presented. PMID- 10829746 TI - Clarification of plasma from whole human blood using ultrasound. AB - There has been interest for a number of years in the possibility of separating blood into cells and plasma by methods other than centrifugation, so that the plasma can be analysed on-line. Cells in whole blood normally occupy about 45% of the suspension volume. It has been shown with a number of different cell types, such as yeast and bacteria, that for concentrations of this order the cells are not as efficiently harvested by ultrasound as those for lower concentrations. In this study, removal of cells from 3-4 ml whole blood volumes has been examined in ultrasonic standing wave fields from tubular transducers driven at a frequency of 1.6 MHz. Samples of whole human blood (n = 11) from two volunteers have been processed by three tubular transducers where high levels of cell removal, 99.7% on average, have been demonstrated with high reproducibility between samples as well as for different transducers. PMID- 10829747 TI - The use of acoustic fields as a filtration and dewatering aid AB - An experimental rig has been developed to study the effects of electric and acoustic field combinations on the filtration rate of titanium dioxide suspensions. Ultrasound energy is applied tangentially to the filter medium. Electric field strengths, suspension characteristics and process parameters can all be varied independently. Results from an experimental programme demonstrate that the use of ultrasound across the cake surface can decrease the specific cake flow resistance and increase the filtration rates of low-concentration rutile suspensions (0.1% v/v). Changes in the conductivity induced by ultrasonic irradiation affect the suspension such that the application of an electrical field is enhanced, giving an equivalent electric field strength higher than that applied. PMID- 10829748 TI - Modelling in the design of a flow-through ultrasonic separator AB - This paper describes the design and testing of a flow-through ultrasonic separation device that allows the concentration of particles within a fluid. The device operates without the use of an acoustically transparent element. Three models are used to examine the behaviour of the cell, dealing with acoustic particle interaction, electro-acoustic characteristics, and fluid flow. The device is able to concentrate up to 84% of the 60 microns sand particles in the 'dirty' stream, 13% in the intermediate stream and 3% in the 'clean' stream. Flow rates of up to 20 lh-1 (equating to an inlet velocity of 10(-2) ms-1) have been used with an electrical power input of up to 50 W (10 kWm-1). PMID- 10829749 TI - Modeling of the acoustic pressure fields and the distribution of the cavitation phenomena in a dual frequency sonic processor AB - An attempt has been made to model the acoustic pressure field and the spatial distribution of the cavitation phenomena in a dual frequency sonic processor. A methodology has been presented with numerical simulations to optimize the conditions of the dual frequency acoustic field. The simulations presented in this work reveal that with manipulation of the parameters (viz., frequency ratio and the pressure amplitude ratio of the two acoustic waves and the phase difference between the two waves) of the dual frequency acoustic field it is possible to control the mode (stable or transient) and spatial distribution of the cavitation events in the sonic processor. It has been shown that two major shortcomings of the sonic reactor, viz., directional sensitivity of the cavitation events and erosion of the sonicator surface can be overcome by application of a dual frequency acoustic field. PMID- 10829750 TI - Dependence of sonochemical luminescence on various sound fields AB - To understand the effect of the sound field on sonochemical luminescence, the exact sound pressure must be determined in each field. In this study it was determined by the Shlieren method, which measures the sound pressure without mixing the sound fields. We compared the efficiency of the sonochemical luminescence in three different ways: changing the diameter of the transducer, combining two transducers to obtain crossed propagating directions and surrounding the sound field by a glass cylinder. In the last case cylinders with various sizes were studied. We found that (i) at the same sound pressure, the larger transducer induces stronger luminescence per unit volume, (ii) driving two transducers produces stronger luminescence than the sum of each transducer and (iii) a glass cylinder surrounding the sound field induces stronger luminescence. PMID- 10829751 TI - Effects of ultrasound on both electrolytic and electroless nickel depositions AB - The effects of ultrasound on both electrolytic and electroless nickel depositions were investigated by polarization and a.c. impedance methods. The ultrasound accelerated the charge transfer process at the metal-electrolyte interface in the electrodeposition and the mass transport process in the electroless deposition. In the electrodeposition with Watts bath, the crystal orientations of deposited film largely changed in the presence of ultrasound. The imposition of ultrasound gave rise to decreasing cathodic overpotential and increasing exchange current density, and these effects depended upon the ultrasonic frequency. The values of exchange current density estimated from a.c. impedance were dependent upon the measured electrode potentials. In the electroless deposition with citrate bath, the deposition rates increased in the presence of ultrasound. There were two kinds of Ni(2+)-citrate complex which were reduced at -0.7 V and -1.1 V. The electroless deposition process was controlled by the Ni(2+)-citrate complex that was reduced at -0.7 V. This reduction rate was diffusion controlled and largely increased in the presence of ultrasound. The effects of ultrasonic frequency on both electrodeposition and electroless deposition increased in order of no irradiation < 100 kHz < 28 kHz < or = 45 kHz. PMID- 10829752 TI - Estimation of the blood Doppler frequency shift by a time-varying parametric approach. AB - Doppler ultrasound is widely used in medical applications to extract the blood Doppler flow velocity in the arteries via spectral analysis. The spectral analysis of non-stationary signals and particularly Doppler signals requires adequate tools that should present both good time and frequency resolutions. It is well-known that the most commonly used time-windowed Fourier transform, which provides a time-frequency representation, is limited by the intrinsic trade-off between time and frequency resolutions. Parametric methods have then been introduced as an alternative to overcome this resolution problem. However, the performance of those methods deteriorates when high non-stationarities are present in the Doppler signal. For the purpose of accurately estimating the Doppler frequency shift, even when the temporal flow velocity is rapid (high non stationarity), we propose to combine the use of the time-varying autoregressive (AR) method and the (dominant) pole frequency. This proposed method performs well in the context where non-stationarities are very high. A comparative evaluation has been made between classical (FFT based) and AR (both block and recursive) algorithms. Among recursive algorithms we test an adaptive recursive method as well as a time-varying recursive method. Finally, the superiority of the time varying parametric approach in terms of frequency tracking and delay in the frequency estimate is illustrated for both simulated and in vivo Doppler signals. PMID- 10829753 TI - Wavelet transforms in estimating scatterer spacing from ultrasound echoes. AB - Ultrasound echoes from organs such as the liver display resolvable periodicity due to regular scattering centers within tissue. The spacing among such scattering centers has been proposed as a signature to characterize diffuse and focal diseases of the liver. Even though it is highly desirable to be able to estimate an inter-scatterer-spacing (ISS) distribution, current methods can estimate only the mean value of scatterer spacing (MSS) over a tissue length. In this paper, we propose a wavelet transform-based technique that is capable of estimating the location of each scattering center, making it possible to obtain the ISS distribution. We represent liver tissue with a point scatterer model, and show, via computer simulations, that the use of multi-scale information in the wavelet scale-space allows us to estimate the locations of regular scattering centers. We show that both the observation noise and random ultrasound returns from unresolvable tissue microstructure can be removed successfully in the wavelet domain via the properties of the modulus maxima sequence of observation across different scales. PMID- 10829754 TI - Detection of bone disease with ultrasound--comparison with bone densitometry. AB - A system for ultrasonic in-vivo examination of a heel bone (calcaneus) was developed. When operating in transmission mode, the system can measure broadband ultrasonic attenuation-BUA, speed of sound--SOS and thickness of bone. BUA and SOS are measured by comparing the pulses transmitted through the heel with the reference pulse transmitted through water. In our approach, we operate in the backscattered mode in addition to transmission. The backscattered ultrasonic technique for bone characterization is very promising because the magnitude of backscattered waves depends on the scattering cross-section of a trabecular structure that, to some extent, describes the microarchitecture of a calcaneus. Additionally, when the backscattered and transmitted signals are compared, some of the signal distortions caused by tissue and bone interfaces are reduced. A set of data representing signals transmitted through the heel and reflected inside a calcaneus for patients with osteoporosis was collected. Several signal-processing techniques were applied in order to smooth the backscattered signal and to calculate a trabecular structure cros-section (TSC) function. Results obtained by these approaches along with a spectral shift method and a standard BUA measurement are presented and compared to X-ray bone mineral density determination results. PMID- 10829755 TI - Improved detection of echo signals from inside bone. AB - The purpose of the study was to obtain useful information for diagnosing internal abnormalities of bones by ultrasound. We fabricated a transmitting and receiving system mainly targeting the bone and an image processing system using a computer, and studied the attenuation of ultrasound in animal bone (in vitro). The average value was about 37 dB. The examination was able to determine successfully the proper dynamic range for detection, and a B-mode image of the inside of the bone could be displayed. Here we describe a new transmitting and receiving system developed to improve the detection of echo signals from inside bones. In this method, two kinds of high sensitivity transducer were made as follows: (1) 5 HMz frequency, 6.5 mm diameter; (2) 10 HMz frequency, 13 mm diameter. By using this method, detectability of the echo signals can be increased more than with the former system. The results showed that the structure of the bones could be clearly seen in the B-mode images. PMID- 10829756 TI - Multiphase nature and structure of biomaterials studied by ultrasounds. AB - The paper discusses the applicability of a two-phase model of saturated porous materials for a description of the results of broadband ultrasonic studies of wave parameters in bovine trabecular bone. The analysis is focused on the role of the internal structure of the materials in the propagation of dilatational waves within the frequency range with a significant attenuation of wave energy due to absorption and scattering. The applicability of ultrasonic studies for the determination of characteristic macro- and micro-structural parameters of biomaterials using a model-based approach is considered. PMID- 10829757 TI - Ultrasonic wave speed measurement using the time-delay profile of rf backscattered signals. AB - Conventional methods determine the ultrasonic wave speed by measuring the medium path length propagated by a pulsed wave and the corresponding time-of-flight. In this study, the wave speed is determined without the need of the path length. A transmitting transducer sends a pulsed wave into the medium (constant wave speed along the beam axis) and the backscattered signal is collected by a hydrophone placed at two distinct positions near the transmitted beam. The time-delay profile, between gated windows of the two rf-signals received by the hydrophone, is determined using a cross-correlation method. Also, a theoretical time-delay profile is determined considering the wave speed as a parameter. The measured wave speed is obtained upon minimization of the RMS error between theoretical and experimental time-delay profiles. A PZT conically focused transmitting transducer with center frequency of 3.3 MHz, focal depth of 20 mm and beam width (-6 dB) of 2 mm at the focus was used together with a PZT hydrophone, 0.8 mm in aperture. The method was applied to three phantoms (wave speed of 1220, 1501 and 1715 m/s) and, in vitro, to fresh bovine liver sample, immersed in a temperature-controlled water bath. The results vary within 3% of those obtained with a conventional method. PMID- 10829758 TI - Improvement of biological activity by low energy ultrasound assisted bioreactors. AB - Bioreactors are broadly applied in biotechnology and wastewater treatments. The so-called advanced bioreactor systems should be optimised in a more compact, more efficient and more effective form. One method of optimisation is the improvement of the solid-liquid interface of the sludge flocs and the mass transfer rate of gas and nutrients in the liquid. This could be obtained by the intake of ultrasound in the bioreactor at a frequency of 25 kHz and a power input of 0.3 Wl 1. An increase in the biological activity of the process investigated with alternating ultrasound as well as a decrease without ultrasound were measured. PMID- 10829760 TI - Extraction and filtering in ultrasonic field: finite element modelling and simulation of the processes AB - In this research, the use of powerful sonification is studied in making the extraction of tannins more efficient. A combined extraction-filtration system using sonification was designed and modelled. Preliminary extraction studies were carried out to show the effect of ultrasound in extraction. In these experiments high-intensity ultrasound was utilised in extraction of tannins of Salix phylicifolia. Different solvents were used in extraction experiments. Pickled sheepskin was tanned with the extracts and with some other vegetable tannins for comparisons. PMID- 10829759 TI - Determination of avocado and mango fruit properties by ultrasonic technique. AB - A nondestructive ultrasonic measurement system was developed for the assessment of some transmission parameters which might have quantitative relations with the maturity, firmness and other quality-related properties of avocado and mango fruits. The system utilizes a set of low-frequency probes arranged to measure the ultrasonic signal transmitted and received over a short distance across the peel. The attenuation of the ultrasonic waves, transmitted through the peel and the attached fruit tissue, changes as a result of the progressive ripening and softening of the fruit during the fruiting season and in the course of storage. The present study quantitatively addressed the linkage between the ultrasonic attenuation and the physiological parameters of the flesh of the fruits. Results were obtained in the time and frequency domain, and the data set was analyzed statistically to identify the relations between the major physiological indices and the ultrasonic parameters. Quantitative relations were developed to describe the linkage between ultrasonic parameters and the maturity, firmness and other quality-related properties in mango and avocado fruits. PMID- 10829761 TI - Use of ultrasound to assess Cheddar cheese characteristics. AB - Blocks of Cheddar cheese were matured in temperature-controlled chambers at 5 and 12 degrees C. The ultrasonic velocity increased during maturation ranging from 1657 to 1677 ms-1 at 12 degrees C and from 1684 to 1693 ms-1 at 5 degrees C. The ultrasonic velocity was related to the square root of the deformability modulus and the slope in puncture. The increase of velocity during maturation shows the feasibility of using an ultrasonic device to non-destructively monitor Cheddar cheese maturity. Ultrasound velocity was measured at different temperatures. The velocity decreased with increasing temperature, and from the slope of the first part of the temperature-velocity curves it was possible to non-destructively assess the moisture content of different types of cheese. PMID- 10829762 TI - Experiences with synthetic aperture focusing technique in the field AB - The detection and evaluation of defects in industrial components relies strongly on ultrasonic inspection techniques. Distance gain size (DG) or reference reflector methods can be improved concerning their localization, signal-to-noise ratio and sizing accuracy by the synthetic aperture focusing technique (SAFT). To obtain a high quality image, parameters like focal probe versus contact technique probe, achieved resolution or features of SAFT images compared with B-scan images are discussed. The implementation of SAFT in a CAD environment allows us to present stacked 2D reconstructions dynamically. On a cladded testblock with half penny shaped cracks the advantage of combining CAD with SAFT is shown. A 3D SAFT example finalizes the overview of two decades of experience in applying this technique. PMID- 10829763 TI - Non-linear algorithms based on SAFT ideas for reconstruction of flaws AB - The main idea and the assumptions of time domain synthetic aperture focusing technique (SAFT) are discussed. It is underlined that the major SAFT assumption is that the flaw can be represented by a number of single non-interacting scalar point scatterers, which act as point sources after illumination with the incident wavefield. As a result some important classes of flaws are incorrectly reconstructed by means of SAFT. Among them are cracks from which reflected signals are similar to those from mirror-shaped flaws. Different non-linear SAFT algorithms proposed earlier by the author are discussed and compared. Among them are SAFT with non-linear normalization (NSAFT), second order SAFT, and the radial tomography technique. The principles of these algorithms are discussed. Then their advantages and disadvantages are analysed. Unfortunately each algorithm cannot be considered universal as a classical SAFT, but for many important cases these algorithms are useful for solving inverse problems in ultrasonics. The ideas of non-linear SAFT algorithms are illustrated by physical experiments. PMID- 10829764 TI - Improvement of the lateral resolution of finite-size hydrophones by deconvolution AB - In various fields of ultrasound applications, frequencies well above 10 MHz are used. As a consequence of this, ultrasound sensors, especially the piezoelectric hydrophones presently available which are used for the characterization of the respective fields, can no longer be considered as point receivers. By means of numerical deconvolution, the adverse averaging effect caused by the finite sensor size can be revoked. The efficiency of the deconvolution process is dealt with for both numerical simulations and experimental investigations. Best results were obtained using a reconstruction filter consisting of a combination of a Wiener filter, a pruning filter and an additional low-pass filter. PMID- 10829765 TI - Experimental investigation of bounded beam reflection from plane interfaces in the vicinity of leaky waves angles AB - The aim of this work is an experimental validation of the reflected and transmitted fields through a viscoelastic material when a bounded ultrasonic beam is incident at different angles. Special measurements are carried out when incidence is performed at some critical angles of the material used. So the propagation of leaky waves along the solid layer is investigated. The inverse procedure for optimizing model parameters from the noisy input-output data is performed using the maximum likelihood estimator. PMID- 10829766 TI - Attenuation and dispersion of Rayleigh waves propagating on a cracked surface: an effective field approach AB - A new effective field approach describing the attenuation and dispersion of a Rayleigh wave propagating on a surface containing a distribution of one dimensional, surface-breaking cracks is presented. Limited by the validity of the independent scattering approximation, the model utilizes the complex transmission coefficient of a single crack to build expressions for the phase velocity and attenuation coefficient of an effective Rayleigh wave. The model is shown to be able to accommodate the effect of compressive residual stresses that tend to close the distributed cracks at their mouth, and therefore substantially reduce the attenuation and the velocity change caused by an equivalent distribution of open cracks. Wherever possible, the predictions of the new model are compared with those of other approaches and critical remarks are offered which discuss the advantages of the effective field approach over the others. Finally, an extension of this approach to distributions of two-dimensional surface-breaking cracks is outlined. PMID- 10829767 TI - Observation of Scholte-like waves on the liquid-loaded surfaces of periodic structures AB - The observation of Scholte-like ultrasonic waves travelling along the water loaded surfaces of solids with periodically varying properties is reported. Results are presented for two 2D superlattices that intersect the surface normally: a laminated solid of alternating 0.5 mm thick layers of aluminium and a polymer, and a hexagonal array of polymer rods of lattice spacing 1 mm in an aluminium matrix. The surface waves are generated and detected by line-focus acoustic lenses aligned parallel to each other, and separated by varying distances. For homogeneous solids, phase matching constraints do not allow the Scholte wave to be coupled into with an experimental configuration of this type, and this is demonstrated with results on a uni-directional carbon-fibre/epoxy composite. These constraints are relaxed for a periodic solid, where coupling takes place through Umklapp processes. In our experiments, the source pulse is fairly broadband, extending up to about 6 MHz, whereas the spectrum of the observed Scholte arrival is peaked at around 4 MHz. We attribute this to a resonance in the surface response of the solid associated with the superlattice structure. On rotating the solid about its surface normal, the Scholte wave displays a characteristic variation in phase arrival time and, to a lesser extent, also group arrival time. This variation is well accounted for with a model that incorporates Umklapp processes in the solid's surface response. PMID- 10829768 TI - Attenuation of ultrasound in porous media with dispersed microbubbles AB - The dispersion relation for a granular bed with a small amount of fine bubbles is formulated and analyzed. It is assumed that the grain size is much larger than the bubble's radius and that their volume concentration is small. The study is motivated by the problem of acoustic diagnostics of fixed bed chemical reactors operating in multiphase flow regime. PMID- 10829769 TI - Calculation and visualization of Lamb wave motion AB - Lamb wave propagation was calculated by using hybrid boundary element method (HBEM) that is the combination of the analytical mode expansion and boundary element method. HBEM makes it possible to simulate Lamb wave propagation in large plates with shorter computational time. The numerical results, e.g. displacements and stresses, were collected at every time step for the inner grid points and visualized as mpeg movies or jpeg pictures. The generation mechanism of Lamb waves is investigated as a numerical example. PMID- 10829770 TI - Micromachined ultrasound transducers with improved coupling factors from a CMOS compatible process AB - For medical high frequency acoustic imaging purposes the reduction in size of a single transducer element for one-dimensional and even more for two-dimensional arrays is more and more limited by fabrication and cabling technology. In the fields of industrial distance measurement and simple object recognition low cost phased arrays are lacking. Both problems can be solved with micromachined ultrasound transducers (MUTs). A single transducer is made of a large number of microscopic elements. Because of the array structure of these transducers, groups of elements can be built up and used as a phased array. By integrating parts of the sensor electronics on chip, the cabling effort for arrays can be reduced markedly. In contrast to standard ultrasonic technology, which is based on massive thickness resonators, vibrating membranes are the radiating elements of the MUTs. New micromachining technologies have emerged, allowing a highly reproducible fabrication of electrostatically driven membranes with gap heights below 500 nm. A microelectronic BiCMOS process was extended for surface micromechanics (T. Scheiter et al., Proceedings 11th European Conference on Solid State Transducers, Warsaw, Vol. 3, 1997, pp. 1595-1598). Additional process steps were included for the realization of the membranes which form sealed cavities with the underlying substrate. Membrane and substrate are the opposite electrodes of a capacitive transducer. The transducers can be integrated monolithically on one chip together with the driving, preamplifying and multiplexing circuitry, thus reducing parasitic capacities and noise level significantly. Owing to their low mass the transducers are very well matched to fluid loads, resulting in a very high bandwidth of 50-100% (C. Eccardt et al., Proceedings Ultrasonics Symposium, San Antonio, Vol. 2, 1996, pp. 959-962; P.C. Eccardt et al., Proceedings of the 1997 Ultrasonics Symposium, Toronto, Vol. 2, 1997, pp. 1609 1618). In the following it is shown how the BiCMOS process has been modified to meet the demands for ultrasound generation and reception. Bias and driving voltages have been reduced down to the 10 V range. The electromechanical coupling is now almost comparable with that for piezoelectric transducers. The measurements exhibit sound pressures and bandwidths that are at least comparable with those of conventional piezoelectric transducer arrays. PMID- 10829771 TI - Air-coupled measurement of plane wave, ultrasonic plate transmission for characterising anisotropic, viscoelastic materials AB - Electrostatic, air-coupled, ultrasonic transducers are used to generate and detect plane waves in viscoelastic, isotropic or anisotropic solid plates. The through-transmitted field is measured and compared to numerical predictions. An inversion scheme is then applied for identifying the values of the complex Cij which are representative of the viscoelasticity properties of the materials. The issue of this work is a contact-free, ultrasonic technique for material characterisation. PMID- 10829773 TI - The measurement and numerical modelling of the diffraction field of a spherical ultrasonic transducer working in the air at the frequency of 1.3 MHz AB - This paper presents the results of measurements of the acoustic diffractive field in the focal area of a spherical ultrasonic transducer working in air at a frequency of 1.3 MHz. The measurements show some differences when compared to the theoretical distribution. Computer modelling of the transducer aperture complex function and the numerical simulation of its radiation are used to explain those differences. It is shown that the wave aberration of threefold symmetry (described by the Zernicke polynomial) and the random distribution of grains in the impedance matching layer are the main causes. PMID- 10829772 TI - Characterization of airborne transducers by optical tomography AB - This paper describes the application of an acousto-optic method to the measurement of airborne ultrasound. The method consists of a heterodyne interferometric probing of the pressure emitted by the transducer combined with a tomographic algorithm. The heterodyne interferometer measures the optical phase shift of the probe laser beam, proportional to the acoustic pressure integrated along the light path. A number of projections of the sound field, e.g. a set of ray integrals obtained along parallel paths, are made in moving the transducer to be tested. The main advantage of the method is its very high sensitivity in air (2 x 10(-4) Pa Hz-1/2), combined with a large bandwidth. Using the same principle as X-ray tomography the ultrasonic pressure in a plane perpendicular to the transducer axis can be reconstructed. Several ultrasonic fields emitted by wide band home made electrostatic transducers, with operating frequencies between 200 and 700 kHz, have been measured. The sensitivities compared favorably with those of commercial airborne transducers. PMID- 10829774 TI - In-line concentration measurement in complex liquids using ultrasonic sensors AB - Recently there has been increased demand for chemical sensors measuring in-line the concentration of selected substances in complex liquids in order to guarantee a high product quality in the process industry. At present there is a great interest in acoustic sensor systems for concentration measurements. This article presents a new ultrasonic sensor system consisting of a miniaturized multi-sensor arrangement for the comprehensive acoustic characterization of liquid mixtures. The sensor system measures sound velocity, impedance coefficient, attenuation coefficient and temperature. PMID- 10829775 TI - Model-based parameterisation of a hydrocyclone air-core AB - An important metric for the accurate control of a hydrocyclone is the diameter of its air-core. Ultrasonic data from a 16-transducer, 1.5 MHz pulse-echo tomographic system are analysed to determine the variation of the air-core diameter with various operating conditions. The back-projection image reconstruction method is not accurate enough for this task. Sub-millimetre accuracy is obtained, however, by applying a combination of signal processing and model-based reconstruction, using the fact that there is a small variation in the air-core boundary position. The findings correspond well to the results obtained from X-ray and electrical resistance modalities. PMID- 10829776 TI - Ultrasonic monitoring of zeolite synthesis in real time AB - Variations of the phase and amplitude of a transmitted ultrasonic wave package were monitored in real time during the synthesis of zeolite A and zeolite X. For both materials, characteristic changes of the measured attenuation and the sound velocity of ultrasonic waves traveling in the reaction fluid were observed, correlating with the processes of gel formation and zeolite crystallization, respectively. Aging effects of the reaction fluids are demonstrated. The observation of the onset of the zeolite crystallization was verified with ex-situ X-ray diffraction (XRD) measurements. PMID- 10829777 TI - Ultrasound process tomography system for hydrocyclones AB - The implementation of a laboratory-based ultrasound tomography system to an industrial process application is not straightforward. In the present work, a tomography system with 16 transducers has been applied to an industrial 50 mm hydrocyclone to visualize its air-core size and position. Hydrocyclones are used to separate fine particles from a slurry. The efficiency of the separation process depends on the size of the air core within the cyclone. If the core is too large due to spigot wear, there will be a detrimental effect on the slurry throughput. Conversely, if the throughput is increased to an extent where the air core becomes unstable or disappears, the particle separation will no longer take place, and the processed batches may become contaminated. Ultrasound tomography presents a very good tool with which to visualize the size, position and movement of the air core and monitor its behaviour under varying input parameters. Ultimately, it could be used within this application both to control the input flow rate depending on the air core size and to detect spigot wear. This paper describes the development of an ultrasonic tomography system applied to an instrumented hydrocyclone. Time-of-flight data are captured by a dedicated acquisition system that pre-processes the information using a DSP and transfers the results to a PC via a fast serial link. The hardware of the tomography system is described, and cursory results are presented in the form of reconstructed images of the air core within the hydrocyclone. PMID- 10829778 TI - Anomalous propagation characteristics of evanescent waves AB - An analysis is done of the crossing of a forbidden region in a thin plate by a backward propagating Lamb wave: the refraction/reflection effects undergone by the coupled modes produced at each boundary of the forbidden region are taken into consideration, as well as the penetration of the backward wave as an evanescent wave. The outcome of the acoustic perturbation is analysed for a few angles of incidence and experiments are performed that confirm the theoretical predictions. PMID- 10829779 TI - Guided elastic waves in a pre-stressed compressible interlayer AB - The propagation of guided elastic waves in a pre-stressed elastic compressible layer embedded in a different compressible material is examined. The waves propagate parallel to the planar layer interfaces as a superposed dynamic stress state on the statically pre-stressed layer and host material. The underlying stress condition in the two materials is characterized by equibiaxial in-plane deformations with common principal axes of strain, one of the axes being perpendicular to the layering. Both materials have arbitrary strain energy functions. The dispersion equation is derived in explicit form. Analysis of the dispersion equation reveals the propagation characteristics and their dependence on frequency, material parameters and stress parameters. Combinations of these parameters are also defined for which guided waves cannot propagate. PMID- 10829780 TI - Location of acoustic emission sources generated by air flow AB - The location of continuous acoustic emission sources is a difficult problem of non-destructive testing. This article describes one-dimensional location of continuous acoustic emission sources by using an intelligent locator. The intelligent locator solves a location problem based on learning from examples. To verify whether continuous acoustic emission caused by leakage air flow can be located accurately by the intelligent locator, an experiment on a thin aluminum band was performed. Results show that it is possible to determine an accurate location by using a combination of a cross-correlation function with an appropriate bandpass filter. By using this combination, discrete and continuous acoustic emission sources can be located by using discrete acoustic emission sources for locator learning. PMID- 10829781 TI - A single plate ultrasonic gyroscope in plane motion AB - We have proposed a vibration gyroscope made of a single piezoelectric plate in plane motion. The gyroscope makes use of the degenerated modes associated with a square plate. In the present paper, the experimental verification using an enlarged piezoelectric ceramic plate is presented, whose results are compared with the numerical prediction based on two-dimensional and three-dimensional finite element modelling. PMID- 10829782 TI - Nonlinear piezoelectricity in PZT ceramics for generating ultrasonic phase conjugate waves AB - We have succeeded in the generation of acoustic phase conjugate waves with nonlinear PZT piezoelectric ceramics and applied them to ultrasonic imaging systems. Our aim is to make a phase conjugator with 100% efficiency. For this purpose, it is important to clarify the mechanism of acoustic phase conjugation through nonlinear piezoelectricity. The process is explained by the parametric interaction via the third-order nonlinear piezoelectricity between the incident acoustic wave at angular frequency omega and the pump electric field at 2 omega. We solved the coupling equations including the third-ordered nonlinear piezoelectricity and theoretically derived the amplitude efficiency of the acoustic phase conjugation. We compared the efficiencies between the theoretical and experimental values for PZT ceramics with eight different compositions. Pb[(Zn1/3Nb2/3)(1 - x)Tix]O3 (X = 0.09, PZNT91/9) piezoelectric single crystals have been investigated for high-performance ultrasonic transducer application, because these have large piezoelectric constants, high electrical-mechanical coupling factors and high dielectric constants. We found that they have third order nonlinear piezoelectric constants much larger than PZT and are hopeful that the material as a phase conjugator has over 100% efficiency. PMID- 10829783 TI - Magneto-acoustic ceramics for parametric sound wave phase conjugators AB - The paper reflects the recent experimental results on the elaboration and study of active materials for magneto-acoustic phase conjugators (MAPCs). The results of complex measurements of MAPC parameters are demonstrated on typical samples of NiFe2O4 magnetostrictive ceramics. The mechanism of strong dispersion of gain increments and output power of MAPCs is studied and explained by dispersion of critical values of the parametric modulation depth of sound velocity. A maximum output power 240 W at frequency 5 MHz is obtained for active element MAPC with critical current Ic = 9 A and electrical Q-factor equal to 80. PMID- 10829784 TI - Acoustic wave scattering from a composed shell reinforced by a single rib: characteristics of peripheral waves AB - The steady-state axisymmetrical problem of a plane acoustic wave scattering from a composed shell is considered. The shell has a cylindrical part and two hemispherical endcaps. The rib is a ring of rectangular cross-section that divides the shell into two equal parts. The motion of the shell and the rib is described by the equations of elasticity theory, and the liquid is described by the Helmholtz equation. The solution is obtained numerically by a coupled finite element/boundary element model. Two peripheral waves are generated in the shell: the membrane S0 wave and the bending type water-borne A wave. The form function, acoustic spectrogram and dispersion curves of the phase velocities are presented, and the effect of the rib on the peripheral waves is discussed. PMID- 10829785 TI - Ultrasonic reflection from randomly distributed cylindrical cavities AB - The propagation of steady-state time-harmonic waves in an elastic solid containing cylindrical cavities that are randomly distributed in a slab region is investigated. When an antiplane wave is normally incident on the cavities, the amplitudes of the coherent reflected and transmitted waves outside the slab are evaluated and plotted as functions of frequency and porosity. PMID- 10829786 TI - Diffraction of light by two ultrasonic beams of frequency ratio 1:2, beyond the Raman-Nath diffraction regime AB - Light diffraction by two ultrasonic waves of frequency ratio 1:2 is examined beyond the Raman-Nath diffraction regime. Measurements were taken of +/- 1 order diffracted light intensities for some Raman-Nath parameter values, as a function of the phase shift between the sound beams. The experimental results obtained are discussed and compared with relevant theoretical predictions. Within the experimental conditions, a quite good quantitative agreement between the experimental data and the appropriate theoretical expectations is observed. PMID- 10829787 TI - Piezoelectric excitation of semiconductor plates AB - The microelectronics revolution is due largely to the electronic properties of semiconducting crystals. Almost all binary semiconductors are piezoelectric, and can be fashioned with high resistivity, permitting incorporation of mechanical motion as an additional variable, leading to the possibility of mixed-effect devices with novel properties. In order to further develop these new structures, this paper provides newly derived expressions for piezoelectric excitation of simple thickness plate modes of binary semiconductors, by both thickness and lateral electric fields. PMID- 10829788 TI - Ultrasonic sensor properties characterized by a PC-controlled scanning measuring system AB - The use of ultrasonic sensors for process control is currently widespread for flow, level or distance measurements. Recently, interest has increased, too in the application of ultrasonic sensors to concentration measurements in complex liquids. In this application there are high demands for a defined and stable quality of the properties of both the sensor transfer function and the sound field characteristic. For a detailed investigation and characterization of ultrasonic sensor propertiess, an efficient PC-controlled measuring system was developed by the Institut fur Automation und Kommunikation (IFAK). In this contribution, this high performance approach is presented to make visible the vibrating ultrasonic sensor surface as well as the sound field in front of acoustic sensors in liquids. PMID- 10829789 TI - Acoustic waves in thin-walled elastic tube with polymeric solution AB - Acoustic waves in a pipe with polymeric liquid are investigated within a quasi one-dimensional approach. Analysis of the dispersion equation has shown that rheological features lead to essential changes in both attenuation and speed of the sound. The results may find application in acoustic rheometry of polymeric liquids and for modeling of fast dynamic processes in polymer production technology. PMID- 10829790 TI - Acoustic scattering from complex elastic shells: visualization of S0, A0 and A waves AB - Acoustic scattering phenomena from complex immersed elastic shells are studied by high-speed Schlieren visualization for spark-generated spherical quasi-Dirac excitations. The situations considered are air-filled cylindrical shells under normal insonification and air-filled cylindrical shells soldered at one end with a hemispherical endcap under axial and non-axial incidences. The results are compared with those provided by the Methode d'Isolement et d'Identification des Resonances (MIIR) method and corresponding analytical theoretical solutions. The combination of these complementary approaches better highlights some of the behaviors of the symmetric S0- and antisymmetric A0-Lamb waves, as well was the Scholte-Stoneley A-wave, on the considered targets. In particular, the influence of the internal solder inhomogeneities on the creation and/or the conversion of these modes is demonstrated. PMID- 10829791 TI - Investigation of magnetic fluids by ultrasonic and magnetic methods AB - The influence of magnetic field on the acoustic and magnetic properties of magnetic liquids is discussed. By fitting the curve of Taketomi's theory to the experimental data of the anisotropy of ultrasonic attenuation, the values of quantities describing the structure of magnetic liquids have been determined. Moreover, the dependence of magnetic susceptibility on frequency has been measured. It shows that two processes of magnetization, based on the mechanisms proposed by Brown and Neel, contribute to the magnetization of the magnetic liquid. PMID- 10829792 TI - The effect of the rate of magnetic field and temperature changes on the ultrasonic wave absorption coefficient in a magnetic fluid AB - The rate of external magnetic field changes as well as temperature influences the structure of a magnetic fluid. This work presents experimental results on changes in the ultrasonic wave absorption coefficient of EMG-605 water-based magnetic fluid as a function of the external magnetic field intensity for different rates of magnetic field changes, at different temperatures. On the basis of these studies, radii of spherical clusters formed in the fluid under the influence of an external magnetic field have been determined. PMID- 10829793 TI - [The 106th Internal Medicine Congress: younger, international, oriented to applicability]. PMID- 10829794 TI - [Dobutamine-induced changes in the myocardial blood flow in patients with coronary heart diseases. A quantitative analysis using [15O] H2O positron emission tomography]. AB - BACKGROUND AND OBJECTIVE: Although dobutamine is currently widely used for stress testing, only little is known about the effects of dobutamine on myocardial blood flow. The purpose of the present study was therefore to analyze quantitatively the regional changes in myocardial blood flow during rest and stress. PATIENTS AND METHODS: In order to assess these effects 17 patients (12 men, five women, mean age 57 +/- 8 years) with symptomatic single vessel coronary artery disease (> 70% stenosis) scheduled for coronary angioplasty underwent dobutamine stress testing with a maximum dose of 40 micrograms/kg/min. Myocardial blood flow was measured using 15O H2O position emission tomography at rest and during maximum stress in ischemic and non-ischemic myocardial regions. RESULTS: Dobutamine stress (median dose 30 micrograms/kg/min) increased the rate pressure product significantly (from rest 8697 [95% confidence interval 7959-9435] to stress 16,512 [15,208-17,815] mmHg/min (p < 0.001). Myocardial blood flow during rest was similar in non-ischaemic and ischaemic regions (0.91 [0.93; 0.83-1.28] vs. 1.10 [1.23; 0.91-1.28] ml/min/g, n.s.). During dobutamine stress myocardial blood flow increased in non-ischaemic regions to 2.17 (2.15; 1.77-2.57) ml/min/g, while myocardial blood flow did not increase in ischaemic regions (1.06 [0.97; 0.83 1.28], p < 0.001). Accordingly, dobutamine coronary reserve was 2.42 (2.55; 2.10 2.74) for non-ischaemic regions and 0.98 (1.05; 0.84-1.13) for ischemic regions (p < 0.001). Rate pressure product and myocardial blood flow were significantly correlated (r = 0.79, p < 0.001). CONCLUSION: A dobutamine-induced increase in rate pressure product was proportional to an increase in myocardial blood flow in non-ischaemic regions. In contrast, myocardial blood flow did not increase in myocardial regions supplied by a severely stenosed coronary artery. PMID- 10829795 TI - [The sonographic picture of an echogenic liver is an indicator of pathologic glucose tolerance]. AB - BACKGROUND: Almost all patients in departments of internal medicine routinely undergo abdominal ultrasound examination in which liver changes indicating hepatic steatosis are often detected. The aim of this study was to assess the occurrence of pathological oral glucose tolerance, manifest diabetes mellitus and other changes indicative of a metabolic syndrome in patients with sonographic signs of hepatic steatosis. PATIENTS AND METHODS: 577 patients were examined during a period of 6 months: 90 patients fulfilled the inclusion criteria. RESULTS: Among the 90 patients with echodense liver who were included in the study (42 female, 48 male) 36 patients presented with previous diagnosis of diabetes mellitus (40%). The oral glucose tolerance test was impaired in 19 patients (21%) and in four patients with manifest diabetes mellitus (4%). Among patients with echodense liver a high percentage presented with obesity and impaired fat metabolism. CONCLUSIONS: The large number of patients with an impaired oral glucose tolerance test converting to manifest diabetes, as well as the large number of patients with manifest disorders of fat metabolism suggest that screening for diabetes should be performed in patients who present with sonographic signs of hepatic steatosis. PMID- 10829796 TI - [Orthotopic liver transplantation in a 33-year-old patient with fulminant hepatitis B and HIV infection]. AB - HISTORY AND CLINICAL FINDINGS: A 33-year-old man with fulminant hepatitis B infection was to have an orthotopic liver transplantation (OLTX) because of the otherwise poor prognosis. During preparations for it he was found also to have an HIV-infection. INVESTIGATIONS: Preoperatively the Quick value was 9%, and he had a grade 2 encephalopathy. His immune status was impaired: 477 CD4-T-helper cells/microliter. Virus load, measured with a quantitative HIV-RNA test, was < 80 copies/ml (i.e. below demonstrable level). DIAGNOSIS, TREATMENT AND COURSE: After the first transplantation had ended in organ failure, a second one two days later was successful. But there were several complications postoperatively: ischaemic toxic tubular renal failure requiring haemodialysis, underperfusion of the right lobe of the liver due to arterial stenosis, pleural effusion, cytomegalovirus infection and cyclosporin-induced hypertension. But all these were successfully managed. At the time of this report, 27 months later, the patient felt well and his immune state was stable. During these 2 years he had a practically normal T helper count and HIV-RNA measured below 80 copies/ml with good liver function. There has been no indication for antiviral treatment and there have been no complications relating to immunosuppression. CONCLUSION: The low life expectancy before effective antiviral drugs are available explains the reluctance to perform OLTX in HIV-infected patients. The favourable course in this case, using highly efficacious combined antiretroviral treatment now being available, indicates that after careful consideration, OLTX can be performed in selected patients with HIV. PMID- 10829797 TI - [Genomics]. PMID- 10829798 TI - [Calcium antagonists more closely examined. A known drug group with novel working principles?]. PMID- 10829799 TI - [Dissection of the anterior interventricular ramus in the setting of a blunt chest injury]. PMID- 10829800 TI - [Hypokalemic thyreotoxic paroxysmal paralysis]. PMID- 10829801 TI - [Death during liver hemorrhage after infarct thrombolysis]. PMID- 10829802 TI - [Therapy of chronic hepatitis C with alpha-interferon in 182 patients. Evaluation of results, response predictive factors and side effects]. AB - BACKGROUND: The aim of this study was to assess the results, predictors of response and side effects of therapy with alpha-interferon (IFN) in chronic C virus (HCV) infection. METHODS: A group of 182 patients (150 chronic hepatitis and 32 cirrhosis) was treated with alpha-interferon--recombinant IFN in 120 cases (66%), and lymphoblastoid IFN in 62 cases (34%)--at a dosage of 3 MU three times weekly for 12 months. All our patients were prospectively followed for at least 6 months, and 133 patients for more than 2 years. RESULTS: A short-term positive response was achieved in 65% of our patients, a sustained response in 34%, and a long-term response in 35%, the responses being similar both for hepatitis and cirrhosis. Liver histology, after 2 years, improved in 73% of long-term responders, whereas it improved only in 34% of relapsers. The predictors of response were: age, duration of disease, baseline levels of gamma glutamyltranspeptidase (GGT) and serum ferritin. Both types of IFN proved to have the same efficacy. Side effects were observed in 52% of our patients, which were correlated with age and female sex. CONCLUSIONS: Interferon therapy yields good results, if administered for 12 months, in young patients with disease of short duration and low baseline levels of GGT and serum ferritin, even in the presence of cirrhosis, if at early-stage. Patients with normal alanine aminotransferase (ALT) levels after 3 years may be considered fully recovered. The dosage employed was well tolerated. PMID- 10829803 TI - [Spectral analysis of EEG signals in Alzheimer's disease]. AB - Alzheimer's disease (AD) represents the most frequent cause of senile dementia in Western countries. The disease has a slow onset and gradual progress. Over the past few years considerable importance has been attributed to the diagnostic techniques of brain imaging able to highlight morphological and functional images. EEG mapping is one of the most widely used methods. METHODS: The authors evaluated the characteristics of EEG mapping in relation to frequency and amplitude in a group of elderly patients with AD and in a group of healthy elderly control subjects. The former group consisted of 10 patients with AD aged between 65 and 83 years old. The control group included 10 healthy subjects who were matched for age and sex. RESULTS: The spectral analysis of EEG signals relating to frequency and amplitude showed a slowing down of underlying activity in AD patients, with a frequency between 1 and 4 c/sec, amplitudes below 5 microW and no arrest reaction. In healthy subjects spectral analysis showed a signal percentage colocated at a frequency of between 8 and 13 c/sec in relation to frequency. CONCLUSIONS: In view of these results, the authors recommend the use of EEG mapping to complete the diagnosis of Alzheimer's disease in the aged. PMID- 10829804 TI - [Sleep disorders in the aged]. AB - OBJECTIVES: The aim of the study was the assessment of prevalence and risk factors of sleep disorders in the elderly. METHODS: The study sample consisted of a group of institutionalized elderly, from two university affiliated community nursing homes in Turin. The following subjects were excluded: a) those aged less than 65 years; b) had length of stay less than thirty days; c) were not able to communicate because of severe aphasia or severe hearing loss and d) had cognitive impairment based on the Short Portable Mental Status Questionnaire (SPMSQ ) > 5. A total of 88 subjects (65-102 ys; 24 male and 64 female) have been studied. Comorbidity was measured using the Acute Physiology and Chronic Health Evaluation scale (APACHE), disability using the index of Activity Daily Living (ADL), depressive symptoms using the Geriatric Depression Scale (GDS); sleep quality was determined using the Pittsburgh Sleep Quality Index (PSQI) and the subjective health by the Self-Rated Health (SRH) was also evaluated. RESULTS: Using the PSQI questionnaire, 70 subjects (79.5%) were identified as "poor sleepers" (PSQI > 5). They complained of difficulties in getting sleep (85.7%) and frequent awakening (75.7%). The most common causes of sleep disturbances were nycturia (51.4%), cough (38.6%) and pain (32.9%). Insomnia correlated with depressive symptoms, poor physical activity, number of medications and bad self-rated health. CONCLUSION: Sleep disorders probably are due to physical, psychological or ambient factors. The identification and correction of treatable causes of insomnia are relevant for improving the quality of sleep and health in the elderly. PMID- 10829805 TI - [Study of mapped slow auditory evoked potentials in Alzheimer's disease]. AB - BACKGROUND: Alzheimer's disease affects around 500,000 persons in Italy and its diagnosis remains essentially clinical. Our aim was to contribute to the diagnosis of Alzheimer's disease by mapping auditory evoked potentials (AEP). Evoked potentials represent the electric response of the CNS to an external stimulus and can be represented as a sequence of waves. Brain mapping allows a chromotopogram to be compiled showing chromatic maps. METHODS: A group of 12 patients were studied aged between 68 and 84 years old who were referred to our attention at the Department of Geriatrics and Gerontology at Catania University and the A.O. Cannizzaro, Geriatrics Unit. All patients completed an assessment form (MM-SE, ADAS) and underwent brain NMR and AEP. RESULTS: These tests enabled us to identify two types of patients (Group A and Group B) based on the responses shown. The overall evaluation of results revealed that patients with Alzheimer type dementia present an anomalous cortical neurogenic response to mapped AEP, and that patients in Group A showed anomalous wave morphology, latency and localisation and asymmetric impregnation. The data gathered in this study did not confirm any specific characteristics of AEP given that similar anomalies are found in metabolic dysfunction syndromes and demyelination pathologies. CONCLUSIONS: However, while it does not constitute a specific technique for screening against Alzheimer's disease, the mapping of AEP may contribute to the diagnosis of this disease. PMID- 10829806 TI - [YKL 40: marker of disease activity in rheumatoid arthritis?]. AB - In patients with rheumatoid arthritis the serum and synovial fluid levels of a glycoprotein called YKL 40 are correlated to the severity of disease. YKL 40 may be related to disease activity in RA; it is induced by vitamine D and is reduced by TGF b. YKL 40 has been isolated from bovine mammary secretions during non lactating period and from Human osteosarcoma cell line MG-63. Some observations on experimental and clinical studies are presented. PMID- 10829807 TI - [Diagnostic procedures in internal medicine. Thoracentesis, paracentesis and arthrocentesis in hospital practice]. AB - The aim of this paper is to give a rapid and concise guide for the diagnostic procedures frequently used in internist's clinical practice. Thoracentesis, paracentesis and arthrocentesis were analyzed as regards to indications, contraindications, procedure of execution and interpretation of findings. Addressed to the young physician, the present paper could also be useful as a rapid guide for the experienced physician which does not frequently use these procedures. PMID- 10829808 TI - Allergic emergencies. A brief introduction. PMID- 10829809 TI - Anaphylaxis and anaphylactoid reactions. Classification and pathophysiology. PMID- 10829810 TI - Allergic emergencies in the physician's office. PMID- 10829811 TI - Exercise-induced anaphylaxis (EIA). AB - EIA is a unique physical allergy with increasing incidence as the exercising population increases. Clinical features are indistinguishable from IgE-mediated anaphylaxis in which the offending allergens are known (food or insect stings). Recognition of the association with exercise is crucial. A wide variety of exercises can induce the symptoms, including brisk walking. Symptoms may not be always reproduced by the same amount and type of exercise in a given patient suggesting that associated factors are also needed. Food is an associated factor recognized with increasing frequency, and in the last 5 yr, wheat has been the most frequently associated. Avoidance of the known associated factors, such as food or nonsteroidals, induces a long-lasting remission of EIA. Treatment does not differ from that of anaphylaxis of any other cause. General recommendations for patients with EIA include avoidance of exercise 4-6 h after eating, avoidance of aspirin and nonsteroidals before exercise, and avoidance of all associated conditions known to trigger attacks in each particular patient. Discontinuation of exercise at the earliest warning symptom is critical. PMID- 10829812 TI - Idiopathic anaphylaxis. The evolution of a disease. PMID- 10829813 TI - Allergic emergencies in the emergency room. PMID- 10829814 TI - Severe acute asthma. PMID- 10829815 TI - Allergic emergencies seen in surgical suites. PMID- 10829819 TI - Healthy lifestyles in the new millennium. PMID- 10829818 TI - Angioedema associated with C1 inhibitor deficiency. PMID- 10829817 TI - Allergic emergencies encountered by the dermatologist. Severe cutaneous adverse drug reactions. PMID- 10829820 TI - The impact of modernisation on the diets of adults aged 20-40 years from Samoan church communities in Auckland. AB - The objective of this study was to describe the food and nutrient intakes of adults in three Samoan church communities located in Auckland, New Zealand. The study had a cross-sectional design and measured usual dietary intake in 437 participants, aged 20 years and over, using a self-completed, 89-item quantitative food frequency questionnaire. Median daily energy and nutrient intakes were calculated and compared by gender and age groups. Men obtained a significantly (p < 0.05) lower proportion of their energy from breads and cereals, fruits and vegetables and fats and oils than women. Compared to those aged 40+ years, those aged < 40 obtained a significantly (p < 0.01) lower proportion of their energy from meat, pulses and eggs, fruit and vegetables, and starchy staples. A significantly (p < 0.01) higher proportion of their energy intake came from takeaways, soft drinks, snacks and dairy products. Nutrient analysis revealed that those aged < 40 years obtained significantly (p < 0.001) more energy from fat and sugar, and less energy from protein (p < 0.001) than the older group. They also had significantly lower intakes, per 1000 kilocalories, of a wide selection of vitamins and minerals. A dietary transition has occurred for New Zealand Samoans, reflecting a shift from traditional to modern dietary patterns and a decline in the nutritional quality of the diet. PMID- 10829816 TI - Reactions to radiocontrast material. Anaphylactoid events in radiology. PMID- 10829822 TI - Diabetes mellitus and associated cardiovascular risk factors in north-east Malaysia. AB - Two thousand five hundred and eight subjects from the state of Kelantan in North East Peninsular Malaysia were included in this study to determine the prevalence of diabetes mellitus and impaired glucose tolerance and their association with cardiovascular risk factors. The overall prevalence of diabetes mellitus was 10.5% and impaired glucose tolerance was 16.5%. There was no difference in the prevalence of diabetes mellitus between males and females but the prevalence of impaired glucose tolerance was higher in females (19.0%) than in males (11.5%). Subjects with diabetes mellitus were more obese (38.4%) than normal subjects (24.1%). They also had a higher prevalence of hypertension (12.9%) and hypercholesterolaemia (71.9%) than normal subjects. Subjects with impaired glucose tolerance also had a higher prevalence of obesity (35.5%), hypertension (9.0%) and hypercholesterolaemia (63.0%) than normal subjects. In conclusion, the prevalence of diabetes mellitus and impaired glucose tolerance was high and they were associated with a high prevalence of obesity, hypertension and hypercholesterolaemia. PMID- 10829821 TI - Gender differences in health-related practices among university students in northeast Thailand. AB - The study aims at appraising gender differences in health-related practices, and health behaviour among university students of the Khon Kaen University, northeast Thailand, using a self-administered questionnaire. Of 539 university students, there were 155 males, 384 females, with mean ages 19.7 (+/- 1.2) and 19.6 (+/- 1.1) years, respectively. Persistent health problems were not significantly different between male and female students (12.9% vs. 15.4%). An average body mass index (BMI) was significantly different between male and female students (20.2 +/- 2.1 vs. 19.8 +/- 2.1). Female students reported positive health habits, in terms of drinking, smoking, sun protection, tooth brushing, fruit consumption, conscious efforts to avoid fat and cholesterol, over men, while regular exercise and safety belt use were more likely practised among male students. Using ANOVA and pairwise multiple comparisons, female students demonstrated better health eating habits than men (p = 0.0001). Coronary heart disease preventive habits between the medical and nonmedical faculty was greater for female students than for male students (p = 0.0006 for gender; p = 0.0001 for faculty). Health behaviour (combination of health eating habits, deliberate nutritional practices and coronary heart diseases preventive behaviour) was found to be better practised among women than men (p = 0.0001). These findings, therefore, should focus attention on university students as a target group for health education. PMID- 10829823 TI - A statewide survey of general practitioners in NSW, Australia, about immunisation and strategies to increase childhood immunisation rates. AB - A statewide survey was conducted to ascertain GPs' views in New South Wales (NSW), Australia, about the potential usefulness of strategies to increase immunisation rates and to facilitate providing childhood immunisation in their practice. The survey also explored the usefulness of information sources about immunisation. From September 1997-January 1998, a cross-sectional study using a four page self-administered questionnaire was undertaken. Four hundred GPs practising in NSW, Australia were randomly selected and 343 were eligible to participate. Of these, 281 returned a completed questionnaire (82% response rate). Ninety-one percent and 88% of GPs, respectively, agreed that television campaigns or registering children with the national Australian Childhood Immunisation Register (ACIR) were likely to increase immunisation rates. Sixty two percent of respondents considered that the media created unwarranted parental concern about immunisation. GPs most commonly rated availability of an ACIR list of children overdue for immunisation, better parent educational material and better access to vaccines as strategies which would make immunisation easier. Sixty percent of respondents felt that increased GP payments would be successful in increasing immunisation rates. Only 51% indicated that they had used the "Australian Immunisation Procedures Handbook 6th edition" (a national clinical practice guideline) in the previous month. This study identified GP support for many initiatives aimed at increasing immunisation rates in Australia although GPs were sceptical about the benefits of some programmes. Studies to monitor the impact of GP incentives on immunisation rates in populations and individual practices are underway. These will be useful in determining whether GPs' opinions found in our study correlate with practice in this regard. PMID- 10829824 TI - Rapid streptococcal testing in Vietnamese children with pharyngitis. AB - Streptococcal pharyngitis has been a significant public health problem in Vietnam for many years. Accurate diagnosis of the infection, however, has been difficult. We carried out a clinical trial of a rapid streptococcal antigen detection test (Quick-Vue (R) Flex Strep A) on a population of 777 children with pharyngitis seen at the Institute for the Protection of Children's Health (Children's Hospital) in Hanoi, Vietnam. Bacterial culture was performed in parallel with the rapid test on simultaneously obtained throat swab specimens. The rapid test was found to be 89% sensitive and 92% specific (96% in children not on prior antibiotics) compared to culture. The test was also found to be convenient and acceptable to patients and clinicians. A significant benefit of the test is that those children found positive are more likely to be treated with penicillin rather than a broad spectrum antimicrobial, which in turn will reduce the likelihood of resistant infections in the future. PMID- 10829825 TI - Pattern and distribution of ocular morbidity in primary school children of rural Delhi. AB - A total of 679 primary school children drawn from developed southern parts and underdeveloped eastern parts of Delhi were examined for ocular morbidity. Their age range was 5-15 years and both sexes had almost equal representation. The prevalence of eye diseases was relatively high. Over 40 percent of all the children studied had one or more ocular problems. Trachoma (18%) was the most common ocular morbidity followed by vitamin A deficiency (10.6%), visual acuity < 6/9 (7.4%) and apparent/latent squint (7.4%). In most of these eye disorders, the majority of the children came from the underdeveloped eastern parts of Delhi. Ocular morbidity seems to be unevenly distributed in rural Delhi with a significantly worse picture in the underdeveloped eastern parts, across the river 'Yamuna'. Though blinding trachoma may cease to be a major problem in India, the country continues to have endemic pockets of its non-blinding form. One puzzling observation was that children with protein-energy malnutrition (PEM) did not show any specific vulnerability to ocular diseases. In fact, those with normal weight height index were having a higher share of eye diseases. Though this issue needs to be explored further in the light of biological plausibility, it appears that children who do not have PEM are also exposed to ocular diseases at least with comparable risk. PMID- 10829826 TI - The Sports Challenge international programme for identified 'at risk' children and adolescents: a Singapore study. AB - The current world wide phenomena of youth suicide which became a major issue for countries in the early nineties, is still growing exponentially. The Sports Challenge program was initiated in 1992 in Western Australia to identify 'at risk' children and adolescents who display: a low sense of basic trust, a sense of shame and doubt, a sense of inferiority and a sense of identity confusion with common characteristics of low self esteem. The subsequent program is based on a strong statistical paradigm encompassing current and historical information with reliable and objective evaluation measures. To this end, since 1992, Sports Challenge has been recognised as a 'World Best Practice' in redressing the issue of 'at risk' children and adolescents. The program now operates in over 150 schools and communities throughout Australia and 24 schools and Detention Centres in Singapore. This paper will allow a window into the development of the program and the successful transfer of the project into Singapore. The Singapore study which began in 1996 has revealed the success of the Sports Challenge program cross culturally with improvement in self esteem and self concept of 'at risk' groups in the range of 18% to 44%. PMID- 10829827 TI - An investigation of the disabled elderly in a geriatric health services facility in an urban area of Japan and attitudes of their family caregiver. AB - We investigated characteristics of 72 clients in a geriatric health services facility (hereafter called GHSF), conditions of their family caregivers, and the factors associated with the caregivers choice of discharge destination. Most of the clients were elderly females with a low degree of independence, and dementia was observed in about 60% of them. The clients had children, but many of them lived alone before admission to the GHSF. The rate of admission from hospitals was high (54%), and that of discharge to hospitals was also high (50%). Sixty seven percent of the clients stayed in for a period of over six months. Most of the family caregivers were daughters or daughters-in-law, and considered themselves to be healthy. Sixty-three percent of them had jobs, and most of the caregivers had no sub-caregiver to assist them. The family caregivers desired the client's home (19.4%), hospital or another GHSF (54.2%), or nursing home (26.4%) as the discharge destination from the GHSF. According to Hayashi's quantification theory type II, the factors related to the home as the discharge destination desired by client's family caregivers are as follows; caregivers used formal home public health nursing visit service before entering the GHSF, the job of the caregiver was a part-time job, the client did not show dementia, the period of care experience was shorter than one year. PMID- 10829828 TI - The prevalence of low back pain in Australian adults. A systematic review of the literature from 1966-1998. AB - The objective of this study was to conduct a systematic review of the literature on the prevalence of low back pain in Australian adults. All Australian low back pain prevalence studies published between 1966 and 1998 were identified. General and methodological criteria using current best practice were applied to each prevalence study. Five studies meeting the inclusion criteria were identified. Out of these, three met the minimum current criteria for methodologic acceptance. These studies were Australian Government Health studies conducted over the past 12 years. However, even these studies were flawed, and thus the true prevalence of low back pain in Australia remains uncertain. A methodologically sound study for Australia is recommended as are best practice guidelines for other studies. PMID- 10829829 TI - Assessment of the 1980-1998 health situation and trends in the Americas, by subregion. PMID- 10829830 TI - Methodological summaries: measuring inequity in health. PMID- 10829831 TI - Norms and standards in epidemiology: case definitions. PMID- 10829832 TI - Creation of the Special Program for Health Analysis (SHA) PMID- 10829833 TI - Floods in Venezuela. PMID- 10829834 TI - Integrated management of childhood illness (IMCI) "Healthy Children: Goal 2002". PMID- 10829835 TI - Health analysis: risks of dying and income inequalities. PMID- 10829836 TI - Case definitions. Diphtheria. PMID- 10829837 TI - Case definitions. Pertussis (whooping cough). PMID- 10829838 TI - Pediatric infectious diseases: future campaigns. PMID- 10829839 TI - Pediatric antibiotic therapy in 1990s. PMID- 10829840 TI - Infants born to HIV-1 infected women: lessons from the past decade. PMID- 10829841 TI - Respiratory syncytial virus infection: clinical presentation and management. AB - RSV infection continues to be a major cause of morbidity and mortality throughout the world. Despite advances in the understanding of its pathogenesis, limited progress has been made in prevention and treatment of RSV infection. Based on the experiences thus far it seems that control of RSV infection will be a difficult and complex task. PMID- 10829842 TI - Listeria Monocytogenes infections. AB - This article reviews current information regarding human infection with Listeria monocytogenes. Significant advances have occurred in regard to our knowledge of the epidemiology, pathogenesis, immunology, and treatment of this disease which was formerly believed to be of importance mainly to veterinarians. It remains a cause of high mortality in the many different groups of compromised hosts it infects unless diagnosis and treatment are rapidly established. PMID- 10829843 TI - Molecular techniques in biomedical sciences: a new era in diagnosis of infectious diseases. PMID- 10829844 TI - Metabolic studies in congenital vitamin D deficiency rickets. AB - Congenital rickets in 3 newborns of mothers with advanced nutritional osteomalacia, healed with maternal breast milk feeding when mothers alone were given calcium supplements and 7.5 mg of intravenous D2 and the mother baby pair protected from sunlight. Maternal plasma biochemistry indicated more severe vitamin D deficiency compared to their newborns (intrauterine foetal priority). The first dose of 7.5 mg of vitamin D3 and calcium supplements to mother healed osteomalacia but did not appear to heal the rickets of their breast fed infants (extrauterine maternal priority for vitamin D). A second dose given at 3 months interval healed the rickets in their infants and the biochemistry of the mother and baby returned towards normal. Congenital rickets developed when maternal bone mineral and vitamin D stores had been completely exhausted. Raised IPTH levels in the newborn suggested that foetal parathyroids were responsive to hypocalcaemic stimulus. PMID- 10829845 TI - Perinatal outcome in relation to mode of delivery in meconium stained amniotic fluid. AB - Meconium staining of the amniotic fluid is a common complication during labour. When facilities like electronic monitoring, foetal blood sampling are not available, it is difficult to decide whether labour should be allowed to continue or caesarean section should be done. Even when caesarean section is done, meconium aspiration syndrome (MAS) can still occur and considerable morbidity and mortality may result in the newborn. Amino infusion is being considered as useful in decreasing MAS and its sequelae. Before resorting to amino infusion, we decided to analyse the perinatal outcome in meconium stained liquor to compare whether early caesarean section offered any advantage. This is a retrospective study of 150 labours complicated by thick meconium stained liquor, during a 12 month period (1992-93). PMID- 10829846 TI - The family attitudes towards febrile convulsions. AB - Giving adequate information about febrile convulsion and its prognosis would be helpful in alleviating parental stress, and would contribute to decrease in the morbidity of febrile convulsion. In this study, the knowledge level of parents on taking body temperature, and decreasing high fever, their attitudes during febrile convulsion and the impact of febrile convulsion on parents are evaluated. One hundred seventy-four parents of 132 children with FC were enrolled in the study. Twenty-seven per cent of parents had no thermometer at home, 32.8% of them did not know how to take a temperature, 72.2% of them did not know the minimum range of increased body temperature, and 69.5% of them did not know how to decrease the increased body temperature. Thirty-six percent of parents recognised the convulsions when their children suffered from them, the others assumed the convulsion were fainting spells (6.9%), near death state (38.5%) and suffocation (18.4%). Thirty-six per cent of parents brought their children to the hospital without doing anything themselves. Most parents (91.4%) had a fear of a recurrence of febrile convulsion in their children. Seventy-four per cent of parents complained of insomnia, 24.3% parents had dyspeptic symptoms even 14 parents had weight loss due to dyspepsia. PMID- 10829847 TI - Psychiatric morbidity in preschool children--a clinic based study. AB - Psychiatric morbidity patterns in preschool children attending a general hospital psychiatric clinic were studied. No psychiatric problem was detected in 48% of the sample, epilepsy being the diagnosis in more than half of these children. Common psychiatric problems were mental retardation (27%), breath-holding spells (11%) and hyperkinetic syndrome (7%). Emotional and behavioral problems, and habit disorders, which are quite common in this age group, were poorly represented in the clinic sample. It appears that mental retardation and epilepsy are the only two main disorders usually perceived by Indian parents as psychiatric problems in preschool age group. PMID- 10829848 TI - Diagnostic and therapeutic evaluation of bronchoscopy. AB - The diagnostic and therapeutic efficacy of bronchoscopy was determined in 85 children. The major indications were foreign body removal in younger patients and evaluation of tracheobronchial pathology in older children. Foreign body was commonly isolated in the toddlers and even in the elder age group (19%) as well as in those with suspected ingestion (44%). Groundnut was the predominant foreign body and right bronchus was the most frequent site. Tracheobronchitis (27%), bronchiectasis (13%) endobronchial tuberculosis (9.4%) and mucus plug (3%) were the other frequent findings. Bronchography was performed in 16 patients and it confirmed the diagnosis in 75% of the cases. Minor complications were encountered in 8% of patients. In the present study bronchoscopy yielded definite results in 83% and in many, including those with normal findings it guided further management. PMID- 10829849 TI - Age at which chest circumference overtakes head circumference in children. AB - Maximum likelihood method with probit transformation was applied to estimate median age at which chest circumference (CC) overtook head circumference (HC) in 1206 children from rural areas of Ballabhgarh in Haryana and in 1505 children from a slum population in Delhi. The results showed that, in case of rural area, this phenomenon occurred at an average age of 31.36 months and in children from urban slums, at an average age of 28 months. In both the areas, this phenomenon occurred at an average age of 20 to 21 months in normally nourished children. In children with grade I malnutrition, this was delayed by about 10 months as compared to those with normal nutrition and in children with grade II or worse malnutrition, this was further delayed by another 9 to 13 months on an average as compared to those with grade I malnutrition. These differences were found to be statistically significant (p < 0.001). These results indicate that the longer the delay in CC overtaking HC, the chance is higher for the children to be severely malnourished. Considering the relative ease of measuring head and chest circumferences, this parameter (age of CC crossing HC) could be useful in assessing the long term trend in the improvement of childhood nutrition in developing countries. PMID- 10829850 TI - Transplacental immunity and waning of maternal antibody in measles. AB - Since transplacental immunity and waning of maternally derived measles specific antibodies play an important role in determining the optimum age for vaccination of infants against measles, a study was carried out in which 150 paired samples and 581 infant serum samples were tested for measles specific antibodies. Out of these paired samples, 132 pairs showed measles antibodies in both mother and cord. HAI antibody was absent in 3 paired samples whereas, 5 mothers could not pass on the antibodies in the cord samples. In the remaining 10 serum samples only cord blood showed the presence of antibodies without the detectable level of antibodies in mother. Statistically no significant difference between the mother and cord blood titers was observed by applying the student 't' test for comparison of the mean (t = 0.01). Analysis of 581 infant serum samples for prevalence of maternal antibodies indicated that 83% of the samples at the age of 3 months or below had measles antibodies but with the increase in age there was tremendous loss with only 19-20% at the age of 6-7 months. After 7 months the percentage of infants which had antibody varied from 11-13%. There was negative correlation between age and seropositivity (r = -0.72) which was highly significant (p < 0.05). PMID- 10829851 TI - Serum alpha feto-protein screening in high risk pregnancies. AB - We assayed alpha fetoprotein (AFP) in serum samples from 2,735 women during 14 to 20 weeks of gestation. Serum AFP levels were elevated in the presence of neural tube defect (NTD) and gut atresia in fetus, twin pregnancies, preterm delivery and neonatal complications. In two of the 23 cases of fetal NTD the diagnosis was suggested by AFP assays, with apparently normal ultrasound findings. Low maternal serum AFP levels were associated with chromosomal abnormalities and hydatidiform molar pregnancy. Calculation of risk for Down syndrome based on maternal serum AFP and maternal age helped to reduce the number of women requiring amniocentesis. Maternal serum AFP assay was helpful in the management of threatened abortion, suspected intrauterine death, and maternal toxoplasma infection. In seven cases where maternal serum AFP was high but ultrasound studies were normal, male babies were delivered. Thus maternal serum AFP assay proved useful in narrowing down the group of women requiring more detailed surveillance and diagnostic studies. PMID- 10829852 TI - Neonatal effects of anesthesia for caesarean section. AB - Seventy eight parturient mothers undergoing elective caesarean section were studied with regard to the immediate neonatal outcome in those receiving general and spinal anesthesia. All mothers were of grade I anesthesia risk, were term and had singleton appropriate for gestational age fetuses. There was no difference in fetal acid base chemistry in the two groups. Babies delivered after general anesthesia appeared relatively depressed requiring more free flow oxygen and bag and mask ventilation, though one minute Apgar scores showed no significant difference in either group. Induction delivery intervals were longer in the spinal group but it was not associated with more morbidity. Uterine incision delivery intervals were very small in both groups and no meaningful conclusion could be drawn as regards effect on the newborn. A plea is made for more frequent use of spinal anesthesia considering its many postnatal advantages. PMID- 10829853 TI - Malakoplakia of colon. PMID- 10829854 TI - Incontinentia pigmenti (Bloch-Sulzberger syndrome): report of a case and review of the Indian literature. PMID- 10829855 TI - Familial growth hormone insensitivity syndrome. PMID- 10829856 TI - The liver: enlarged or pushed down? PMID- 10829857 TI - Breastfeeding in the low socio economic group--a matter for concern. PMID- 10829858 TI - Assessment of clinical competence. PMID- 10829859 TI - Metaphyseal chondrodysplasia: a differential diagnosis of rickets. PMID- 10829860 TI - Which sleep position is best for infants? PMID- 10829861 TI - CEPH contribution to the human genome mapping. PMID- 10829862 TI - Organization of neonatal services in developing countries. PMID- 10829863 TI - Micronutrient deficiencies in children--the problem and extent. PMID- 10829864 TI - Micronutrient deficiencies: public health implications. PMID- 10829865 TI - Assessment of zinc status in man. PMID- 10829866 TI - Zinc supplementation in the treatment of childhood diarrhoea. PMID- 10829867 TI - Vitamin A, immunity and infection. PMID- 10829868 TI - Acute chest syndrome in children with sickle cell disease. AB - Acute chest syndrome (ACS) is an acute pulmonic process in patients with sickle cell disease. We prospectively studied 50 patients with ACS admitted to the Pediatric Medical Ward during one year period (Jan. 1993 through Dec. 1993). Twenty eight of them were males and twenty two were females giving a male: female ratio of 1.2:1. The age ranged between one and 12 years. Twelve (24%) of the patients had chest pain on presentation. Twenty seven (54%) patients had significant temperature (> 38 degrees C). The x-ray findings showed that the right lung was involved in 30 patients, the left in 10 patients and both lungs in 10 patients. Three patients had pleural effusion that required chest tube insertion. Laboratory profiles showed that the erythrocyte sedimentation rate ranged between 15 and 90 mm/h, and their hemoglobin ranged between 4.2 gm and 12 gm/dl. Seven (14%) patients had significantly positive mycoplasma pneumoniae titer. None of the blood cultures was positive. All of our patient received antibiotic, usually either Cefuroxime or Ceftriaxone with Erythromycin in addition to other supportive measures such as blood transfusion, oxygen therapy and hydration therapy. PMID- 10829870 TI - Knowledge and attitude of the Bangladeshi rural mothers regarding breastfeeding and weaning. AB - Two hundred and forty two mothers in 7 villages of Narayanganj district, Bangladesh were interviewed to assess their knowledge and attitude regarding breastfeeding and weaning using pretested questionnaires, though 83.5% mothers knew that colostrum is good for the child, less than 8 percent of them gave it as the first food to their babies. Most mothers did not have the correct knowledge about exclusive breastfeeding and the appropriate time for introduction of weaning foods; and only 3% of them knew how to prepare proper weaning foods. The mean score of knowledge of the mothers was only 4 +/- 1.7 out of 10, indicating the need for nutrition education in this area. PMID- 10829869 TI - Nerve injuries due to obstetric trauma. AB - The incidence of nerve injuries among 32,637 deliveries over a period of ten years was 1.81/1000. Brachial plexus injury (1/1000) and facial nerve injury (0.74/1000) accounted for 98% of nerve injuries. Both the right and left side were involved equally. Bilateral nerve injury was not seen. Lack of antenatal care, macrosomia, abnormal presentations, and operative vaginal deliveries significantly increased the risk of nerve injuries. These babies had significantly higher incidence of meconium stained liquor and intrapartum asphyxia. Parity of the mother, gestational age and sex of the baby did not have significant role in the causation of nerve injuries. Injuries to brachial plexus and facial nerve were seen even in babies born by caesarean section, when it was performed for obstructed labour caused by cephalo-pelvic disproportion and abnormal presentations. Three babies with injuries expired and forty-three could be followed up for varying periods. None of the babies had residual defects. Detection of cephalopelvic disproportion and abnormal lie in the third trimester and their appropriate management would decrease the incidence of obstetric palsies to a significant extent. PMID- 10829871 TI - Anesthesia for caesarean section and immediate neonatal outcome. AB - Seventy-eight parturient mothers undergoing elective caesarean section were studied with regard to the immediate neonatal outcome in those receiving general and spinal anesthesia. All mothers were of grade I anesthesia risk, were term and had singleton appropriate for gestational age babies. There was no difference in fetal acid base chemistry in the two groups. Babies delivered after general anesthesia appeared relatively depressed requiring more free flow oxygen and bag and mask ventilation though one minute Apgar scores showed no significant difference in either group. Induction delivery intervals were longer in the spinal group but it was not associated with more morbidity. Uterine incision delivery intervals were very small in both groups and no meaningful conclusion could be drawn as regards effect on the newborn. A plea is made for more frequent use of spinal anesthesia considering its many postnatal advantages. PMID- 10829872 TI - Law-like relationship between height and weight of school children. AB - Ehrenberg's Law-like relationship between height and weight [log (w) = ah + b] was applied to a group of public school and Govt. school children in New Delhi. It was found that the model log (w) = 0.8 h + 0.4 was comparatively a good fit for public school children and the model log (w) 0.8 h + 0.35 for Govt. school children. On fitting the model to the sample data it was found that the value of 'a' was 0.8, with some minor variation in the second decimal point and the value of 'b' was 0.39 & 0.36 respectively for public school boys and girls and 0.32 & 0.29 respectively for Govt. school boys & girls. These results corroborates with the findings of the earlier studies that the intercept constant 'b' in the Law like relationship equation is influenced by the nutrition of children. This model is recommended because of its simplicity and practical usefulness using only already available information. PMID- 10829873 TI - Congenital nephrotic syndrome with clinical hypothyroidism. AB - A 15 month old boy with typical features of congenital nephrotic syndrome (CNS) is reported, who in addition to the renal pathology had an associated clinical hypothyroidism with low levels of total and free thyroxine and triiodothyronine and an elevated serum TSH. Improvement in the physical parameters and mental status from thyroid hormone replacement therapy is documented. PMID- 10829874 TI - Is vernix caseosa a protective material to the newborn? A biochemical approach. AB - Twenty random samples of vernix caseosa were collected from immediately born neonates, in Jamahiriya Hospital, Benghazi. Biochemical studies of these samples revealed presence of lipids (62.5%), proteins (36%) and carbohydrate (1.5%). Also we could observe inhibition of staph. aureus and klebsiella growth on nutrient agar by this vernix. This observation could be explained either by its higher asparagine content or by its elevated lipid component. In addition tripalmitin was found to be the major lipid constituent, responsible for its hydrophobic property. So we recommend leaving this vernix layer on newborn skin until spontaneous drying. PMID- 10829875 TI - Late presenting gangrene of the extremity in an infant of a diabetic mother. PMID- 10829876 TI - Familial growth hormone insensitivity syndrome. PMID- 10829877 TI - Inherited 11q partial trisomy. PMID- 10829878 TI - Recovery from bilateral renal vein thrombosis on supportive management alone. PMID- 10829879 TI - Vaccines for tomorrow. PMID- 10829880 TI - Bacterial meningitis in children. PMID- 10829881 TI - Bacterial toxins in pediatric infectious diseases. PMID- 10829882 TI - Pneumonias in newborn babies. PMID- 10829883 TI - Epidemiology and prevention of AIDS in children. PMID- 10829884 TI - Study of congenital duodenal obstruction. AB - A study of 23 neonates with congenital duodenal obstruction is reported. Their mean gestational age was 38 weeks and mean weight was 2.2 kg. Main clinical features observed were vomiting (100%), which was bilious in 74%, and epigastric fullness with visible peristalsis (74%). Plain X-ray abdomen confirmed the diagnosis in 78%. Associated congenital malformations were seen in 39% of cases. Eleven babies had an intrinsic defect, 11 had extrinsic defect and one baby had combination of intrinsic and extrinsic defect. Malrotations along with band was seen in 39% of cases. Reported mortality was 39%. PMID- 10829885 TI - Influence of nutritional and haemoglobin status on malaria infection in children. AB - Anaemia and malnutrition have been suggested to confer some degree of protection against malaria infection. Therefore, the influence of nutritional status as assessed anthropometrically and that of haemoglobin level on the incidence of malaria and the degree of parasitaemia was studied in 330 children in the age range of 1-9 years in Bisra block near Rourkela in Orissa state. Moderate to severe malnutrition as assessed from percentage of ideal weight was found in 48.8% (161/330) of children but only 8.8% (29/330) of children had some degree of malnutrition when assessed as weight for height indicating presence of chronic malnutrition. Similarly, 197 children (59.7%) had various grades of anaemia as estimated by haemoglobin level prevalence of malaria on initial examination was 27.6% (91/330), while incidence of clinical malaria in children during the year of study was 250/1000. There was no statistical difference in the Prevalence/incidence of malaria or severity of P. falciparum parasitaemia at different haemoglobin or nutritional levels (p > 0.05). No child died or developed severe complications requiring hospitalization. Therefore, anaemia or malnutrition do not appear to provide any protection against malaria or degree of parasitaemia in the children around Rourkela. PMID- 10829886 TI - Do neonates with meconium aspiration syndrome require antibiotics? AB - A randomized clinical trial was conducted to evaluate the utility of antibiotics in the routine management of Meconium Aspiration Syndrome (MAS). Neonates diagnosed to have MAS were randomly allocated to either the antibiotic group (n = 20) receiving gentamicin for 7 days, or the control group (n = 20), receiving no antibiotic. All infants were given identical supportive care. The two groups were comparable with respect to birth weight, gestation, sex distribution, mode of delivery, Apgar scores, and initial clinical and radiological severity of the illness. Results showed that the mean duration and the severity of respiratory distress at 24 hours and 48 hours were similar in the two groups. No secondary infection was documented in either group. A single death occurred in the antibiotic group. It is concluded that empirical use of antibiotics in the routine management of meconium aspiration syndrome is of no benefit. PMID- 10829887 TI - Maternal determinants of birth weight of north Indian babies. AB - Birth weight charts of uncomplicated singleton pregnancies (n = 3293) were prepared from subjects who delivered between 30 and 42 weeks of gestation. Multiple regression analysis of the data revealed significant effect of maternal age, parity, height, weight and religion upon birth weight. Maternal blood pressure systolic > or = 160 mm of Hg and diastolic > or = 110 mm of Hg (severe PIH) were associated with significant lowering of birth weight. Male babies weighed more than female babies. PMID- 10829888 TI - Enteric viral infections in pre-school children in Karachi, Pakistan. AB - A prospective study was conducted in Karachi, Pakistan on the virology of enteropathogens excreted by children with acute gastroenteritis and the results were compared with a control group of healthy children. Rotavirus and Adenovirus detection was done using ELISA techniques, while enterovirus isolation was done by virus culture. In 1990, 12.3% children with acute watery diarrhoea excreted rotavirus, as compared to 24.4% children in 1991. None of the healthy children excreted adenovirus 40 and 41. Preliminary results of 1992 revealed that rotavirus was seen in 13% of children with acute watery diarrhoea and adenovirus in 10% of children. Enteroviruses were isolated in the same frequency in all three groups i.e. children with acute watery diarrhoea, children with poliomyelitis and healthy children. Non-polio enteroviruses were excreted in 50 52% in all the 3 groups. The rate of enterovirus excretion is much higher than seen in other developed countries and is the same in children with diarrhoea and healthy children. PMID- 10829889 TI - Hyperglycemia in malnourished children with dehydrating gastroenteritis. AB - Fifty patients with grade III and IV malnutrition with diarrhoeal dehydration were studied. Hyperglycemia was present in 5 cases at admission. Serum sodium levels were normal in all of them except one case who had hyponatremia. Blood sugar levels were normal in all the cases after rehydration with ORS. Possible mechanisms in pathogenesis of hyperglycemia are discussed. PMID- 10829890 TI - A study on the health status of twin babies. AB - A study on twin births was conducted from May 1993 to April 1994 at S.A.T. Hospital, Thiruvananthapuram. The twinning rate in this period was found as 17.33 per 1000 births. 79.6% babies in the twin pairs were of like sex and 20.4% were of unlike sex. The incidence of low birth weight twin babies in this study was 68.9%. There was no significant difference in the physical parameters (birth weight, body length and head circumference) of twin babies in relation to sex. The incidence of twin birth was higher among primipara mothers and in the mothers of age group 21 to 25 years. The first born baby (twin A) was found to be heavier than the second born baby (twin B) in 44.78% twin pairs and their mean birth weight difference was 438 g, while the second born baby (twin B) was found to be heavier than the first born (twin A) in 38.31% twin pairs and their mean birth weight difference was 291 g. Statistical analysis showed that the difference between the mean birth weight difference in the case of twin A > B was significantly greater when compared to that of twin A < B. PMID- 10829891 TI - Poncet's disease: tuberculous rheumatism. PMID- 10829892 TI - Familial dysautonomia (Riley-Day syndrome). PMID- 10829893 TI - Fetal malformations caused by amnion rupture. PMID- 10829894 TI - Giant congenital nevus. PMID- 10829895 TI - Strategies for combating anemia in adolescent girls: from the present to the future. PMID- 10829896 TI - Panic after measles vaccination: who is to blame? PMID- 10829897 TI - Review of experiences with iodized oil in national programmes for control of iodine deficiency disorders. PMID- 10829898 TI - Ventilatory support for infants in emergency and in the intensive care unit. AB - Pediatric anesthesia and intensive care management has improved dramatically over the past two decades. Improved understanding of the pathophysiology underlying newborn surgical emergencies, new medications and new modes of ventilatory support have all contributed to better patient outcome. The authors have reviewed the anatomy and physiology of the infant airway, indications for and principles of endotracheal intubation, the management of newborn surgical emergencies, indications for post-operative ventilatory support, different modes of mechanical ventilation available, complications of mechanical ventilation with weaning parameters and extubation criteria. The introduction of nitric oxide and the implications of extracorpreal membrane oxygenation in the management of newborn emergency refractory to conventional ventilation are discussed. PMID- 10829899 TI - Psycho-social aspects of childhood AIDS in developing countries. PMID- 10829900 TI - Vitamin A status of socio-economically backward children. AB - A comprehensive survey was carried out to asses the Vitamin A status of pre school (0-6 yrs). and school age (6-12 yrs.) children of socio-economically backward families from slums of Bombay and its suburbs. The Vitamin A, protein, calories and iron from the rice and dal based diet was found to be below recommended dietary allowances (RDA). Among the 1956 children surveyed 20% of the children showed low (< 20 micrograms/dl) serum vitamin A levels. 4.8% of the children were suffering from one or the other signs of Vitamin A deficiency. Rose Bengal stain test (RBST) and conjuctival impression cytology (CIC) indicted the signs of mild conjuctival xerosis and of early epithelial changes which were correlated with serum vitamin A levels. Serum iron, PCV, Hb and RBC levels were below normal. The anthropometric measurements of these children were below 50th percentile of Indian Council of Medical Research (ICMR) standards. Due to lack of proper nutrition, the overall growth of children is either retarded or not upto the standard levels as was noted in majority of the children. PMID- 10829901 TI - Drug therapy of acute diarrhoea in children--actual practice and recommendations. AB - Forty eight private medical practitioners and fifty six pharmacists were interviewed, using a prepared questionnaire in each group, in an attempt to evaluate the awareness and practice in the treatment of acute diarrhoea in children among private medical practitioners and the role of pharmacists in the dispensing of drugs for diarrhoea in children. 83% of the doctors prescribed an antibacterial agent, 56% prescribed loperamide, 19% of them prescribed diphenoxylate plus atropine while 31% prescribed an absorbent in children for the treatment of acute diarrhoea. Of fifty six pharmacists interviewed, 30 of them sold diphenoxylate on their own and 26 of them honoured a doctor's prescription of the same. 80% of pharmacists interviewed dispensed loperamide, more than 50% of them dispensed an antibacterial agent while 14% dispensed a binding agent such as pectin or kaolin for the treatment of acute diarrhoea without a valid prescription from a doctor. We recommend that the pharmaceuticals prominently display the harmful effects in children of drugs such as the antimotility drugs and immediately withdraw from the market, all pediatric preparations of the same. PMID- 10829902 TI - Awareness of and attitude towards oral rehydration therapy. AB - Forty eight Medical practitioners, 56 pharmacists and 55 mothers of children with diarrhoea were interviewed, using a prepared questionnaire, in an attempt to evaluate in each group, the awareness of and attitude towards oral rehydration therapy. 69% of mothers interviewed were aware of oral rehydration therapy, but only 66% among them practised it. 48% of mothers interviewed, received advice regarding oral rehydration therapy from medical personnel and health care providers. It was found that the ORS preparations not conforming to WHO formulation were the most widely stocked and used and the most widely sold. 9% of pharmacists advocated only glucose as a treatment for diarrhoea on their own. 60% of doctors advised and prescribed ORS preparation not conforming to WHO formulation, 44% of medical practitioners were not sure of the exact method of dilution and preparation of the commercially available preparations. The study has shown that there is more to be done towards realising the total success of ORT. We recommend that ORS preparations not conforming to WHO formulation be withdrawn from the market and further that the responsibility of their manufacture be undertaken by governmental agencies. PMID- 10829903 TI - An outbreak of multidrug resistant typhoid fever in Bangalore. AB - Six hundred and eighty five blood cultures from children clinically diagnosed as enteric fever yielded 176 salmonella strains showing isolation success rate of 25.7%, S. typhi were 164 (93.2%), S. paratyphi A 5 (2.8%), S. choleraesuis 4 (2.3%) and S. typhimurium 3 (1.7%). Antibiogram of 164 isolates of S. typhi showed triple drug resistance (TDR) in 156 strains (95.1%) to chloramphenicol, ampicillin and cotrimoxazole, and sensitivity of 90.2% and 95.1% to norfloxacin and ciprofloxacin respectively. Minimum inhibitory concentrations (MIC) of chloramphenicol were between 360 mcg and 640 mcg per ml. Phage types of 38 strains of TDR S. typhi were predominantly E1 and 0 with prevalences of 47.4% and 36.8% respectively in this region. All children with S. typhi isolates sensitive to quinolones in Vitro responded well to these drugs with almost no relapse and hence, the newer generation of quinolones could be considered as the first choice in the primary treatment of enteric fever. PMID- 10829904 TI - Exclusive breastfeeding: protective efficacy. AB - A longitudinal study of feeding practices of and morbidity in 537 infants was undertaken. Feeding practices were assessed at monthly follow-up visits. All infants were initially exclusively breastfed but their percentage dropped to 59.8% and 35.3% at the end of 3 months and 6 months respectively. Exclusively breastfed babies were three-times less likely to fall sick than artificially fed babies. Exclusive breastfeeding was also associated with significantly lowered rate of serious illnesses as shown by fewer rate of hospitalisation (0.52/100 children months vs 4.5/100 children months). Premature introduction of supplementary feeding diluted the protective effects of breastmilk. PMID- 10829905 TI - Effect of birth asphyxia on serum calcium levels in neonates. AB - Serum calcium and phosphorus levels were measured at birth, 6 hours, 24 hours, and on 5th day of life in 35 neonates with birth asphyxia (one-minute Apgar score of 6 or less), and in 37 neonates without asphyxia (one-minute Apgar score of 7 or more). Infants were divided into three groups: FT-AGA (n = 30, asphyxia = 15), FT-IUGR (n = 20, asphyxia = 10) and PT-AGA (n = 22, asphyxia = 10). Asphyxiated infants--FT-AGA as well as FT-IUGR--had significantly lower serum calcium levels than control infants during each of the time period studied. In PT-AGA infants with asphyxia, the serum calcium was significantly low only on 5th day of life. Lack of calcium intake, and hyperphosphatemia were identified as possible risk factors for low serum calcium in asphyxiated infants. No change in serum calcium levels was found in bicarbonate-treated asphyxiated infants in comparison to those who did not receive sodium bicarbonate. In view of the high incidence of low serum calcium in asphyxiated infants, serial monitoring of serum calcium levels is recommended in these infants. PMID- 10829906 TI - Prevalence of tobacco use among school and college going adolescents of Haryana. AB - The prevalence of tobacco use among the school and college going adolescents of Haryana, a state in northern India was studied. 1130 male and 256 female students were given a self administered questionnaire regarding tobacco use. 160 (14.2%) male and 6 (2.3%) female students reported to have smoked at any time in the past. The prevalence of current smoking (defined as smoked in last one week) was only 7.1%. Of the 166 ever smokers, 44% had started this habit between 10 to 15 years of age. There was no rural-urban difference. Univariate analysis showed, male sex, age > 15 years, smoking by father, mother, sister, and friends to be significantly associated with smoking by the adolescent children. By logistic regression, only smoking by friend (O.R. 6.14), brother (O.R. 1.74) and male sex (O.R. 4.67) were found to be significantly associated with smoking by adolescent children. About 8% of the smokers were spending more than Rs. 10 per week on this habit. Both the smokers and non smokers were well aware of the adverse health effects of smoking indicating that mere provision of information on hazards of smoking may not be enough to reduce the prevalence of smoking. A more concerted effort for inducing behavioural change including legislation, communication is called for the fight against smoking. PMID- 10829907 TI - Coping and adaptation in acute lymphoblastic leukemia. AB - The coping and adaptation of 39 children (6-12 years) to Acute Lymphoblastic Leukemia was studied during the first remission with a projective technique- Children's Apperception Test (CAT-S). Seventy seven percent of the children gave adequate responses. Among these, disease awareness was present in 96%, expectation of a favourable outcome was held by 70%, though 61% evidenced emotional distress. Children's psychiatric morbidity was found to be associated with non-response to CAT-S, to anxiety related themes, and negatively with ability to maintain an expectation of a positive outcome. PMID- 10829908 TI - Hematological manifestations of congenital cytomegalovirus infection. AB - Nine infants suffering from congenital cytomegalovirus infection were studied for clinical features with special reference to hematological manifestations. The mean age of appearance of first clinical symptom of disease was 14 days and mean age at presentation was 58 days. Significant pallor, thrombocytopenia and evidence of hemolysis were present in 8 (89%), 4 (44%) and 4 (44%) patients respectively out of three bone marrow examination performed, paucity of erythroid and megakaryocytic cells were seen in two. The other clinical features included hepatomegaly in a (100%) splenomegaly in 6 (66%) each, petachial rash in 5 (55%), Hepatitis in 4 (44%) optic atrophy and corneal opacities in one patient each. Two patients died. Remaining patients showed symptomatic improvement without specific therapy. PMID- 10829909 TI - GM2 gangliosidoses: a review of cases confirmed by beta-N-acetylhexosaminidase assay. AB - The inborn errors of GM2 ganglioside metabolism cause GM2 ganglioside to accumulate within the lysosomes of the nerve cells. The majority of the patients are infants with the Tay-Sachs form of the disease associated with a severe deficiency of beta-N-Acetylhexosaminidase A (hexosaminidase A). Both Hexosaminidase A and B are deficient in Sandhoff disease. The serum total hexosaminidase and the percentage of hexosaminidase A and B were estimated in 449 patients who presented with progressive mental-motor retardation. Three cases of Tay-Sachs disease and two cases of Sandhoff disease were detected. They presented with exaggerated startle response to acoustic stimuli, seizures, optic atrophy and retinal cherry red spots in addition to psychomotor retardation. One case of Sandhoff disease had hepatosplenomegaly and skeletal deformities. PMID- 10829910 TI - Enzyme studies in GM2 gangliosidiosis, and their application in prenatal diagnosis. AB - Assay of hexosaminidase A and B enzymes in four cases with developmental regression and cherry red spot on fundus examination confirmed that three cases had Tay-Sachs disease, and one case had Sandhoff disease. Prenatal diagnosis was carried out by hexosaminidase enzyme assay in amniotic fluid and cells in one family, and chorionic villus sample in the second family. The fetus was diagnosed to be unaffected in one, and affected in the other family. Assay of hexosaminidase A and B is useful for specific diagnosis of GM2 gangliosidosis, and for prenatal diagnosis to reduce the burden of these disorders. PMID- 10829911 TI - Hyperuricemia and acute renal failure in a newborn. PMID- 10829912 TI - Heat stroke--a probable cause of multiple fetal anomalies. PMID- 10829913 TI - Oculopharyngeal muscular dystrophy. PMID- 10829914 TI - Rett syndrome--familial recurrence. PMID- 10829916 TI - Pediatric hepatology. PMID- 10829915 TI - Pedigrees of patients with malignancy. PMID- 10829918 TI - Metabolic liver diseases in childhood. PMID- 10829917 TI - Hepatitis C in childhood. PMID- 10829919 TI - Autoimmune hepatitis. AB - Autoimmune hepatitis can present as either acute or chronic disease in children. Clinical and laboratory features, including association with extrahepatic autoimmune syndromes and prompt response to immunosuppressive treatment, circulating autoantibodies and hypergammaglobulinemia, suggest an immune etiology. However, the disease mechanism remains uncertain. Different types of autoimmune hepatitis are defined on the basis of which autoantibodies are present: anti-smooth muscle (type 1), anti-liver/kidney microsomal (type 2), or anti-soluble liver antigen (type 3). Diseases which may be clinically similar to autoimmune hepatitis must be excluded before the diagnosis of autoimmune hepatitis is established: Wilson's disease, primary sclerosing cholangitis, chronic hepatitis B or C, and drug-induced liver disease are among the most important entities. Corticosteroids alone or with azathioprine constitute the usual treatment for autoimmune hepatitis. Although some children achieve a complete remission, or even recovery, and can stop immunosuppressive treatment, others required low-dose prednisone treatment indefinitely. PMID- 10829920 TI - Nutrition and growth in patients with chronic liver disease. AB - Protein energy malnutrition leading to growth failure is an inevitable consequence of chronic liver disease in childhood. Although the precise pathophysiology is not understood considerable progress has been made in understanding the mechanisms of fat malabsorption and protein turnover in liver disease. There are many difficulties with the correct assessment of nutritional parameters in children with liver disease related to their abnormal body composition and energy expenditure and care needs to be taken with the interpretation of results. The effects of malnutrition secondary to chronic liver disease are varied and include fat soluble vitamin deficiencies, generalised growth failure, impairment of gastrointestinal function, immunosuppression and hypotonia. It is now recognised that malnutrition is an important risk factor for liver transplantation and increases both mortality and morbidity. Strategies to prevent or reverse malnutrition are now established and include the use of specific infant formulas based on low salt protein and an increased concentration of medium train triglyceride (50-70%). Careful nutritional support in association with generous fat soluble vitamin supplementation may produce dramatic improvement in catch up weight gain but for those children in whom growth failure persists, the only management is liver transplantation. PMID- 10829921 TI - Towards the elimination of iodine deficiency disorders in India. AB - Iodine deficiency disorders (IDD) are an important public health problem in India with an estimated 270 million people at risk of IDD. India has adopted the strategy of salt iodization for control of IDD and has the goal of "Universal iodization of salt by 1995 and elimination of IDD by 2000". There is a high degree of political commitment which need to continue if the goal is to be achieved. Currently the ban on ale of un-iodized salt is only applicable to salt on human consumption. There is a need for extending the ban to include salt for animal consumption as IDD affects livestock as well. India has the installed capacity to produce its requirement of 5 million tonnes iodised salt. Communication strategies have to be strengthened especially to educate people who have concerns about of iodine toxicity. The success to a large extent depends on the quality control and monitoring of iodine content of salt at all stages from production to consumption. NGO's and the community have to be encouraged to participate in this process. To sustain the elimination of IDD, a partnership of various stakeholders IDD elimination is essential. PMID- 10829922 TI - Management of tuberculosis. AB - Tuberculosis is a common disease in children but it is highly mismanaged. It is bound to be triggered off by HIV infection in the near future. The importance of HIV and TB has become apparent from the high incidence of disease caused by mycobacteria in AIDS patients. The injudicious use of needles and syringes should be discouraged in high HIV prevalence countries. Multiple drug resistant tuberculosis is emerging as one of the most important problems in medical history. It is often created by health workers who administer anti-TB drugs improperly. The diagnostic aspects and treatment are discussed. PMID- 10829923 TI - Tubercular lymphadenitis: clinical manifestations. AB - One hundred and thirteen children suffering from tubercular lymphadenitis proven histopathologically, were studied for clinical and laboratory features. Age distribution was equal in all age groups except during infancy when it was rare. Sex ratio indicates a male preponderance with male to female in the ratio of 1.5:1 (67:46). Family history of contact with known tuberculous patient was positive in 19 (17%) children, 44 (40%) had received BCG, and 98 (88%) were either normal nourished or had mild malnutrition. Cervical, axillary and inguinal nodes were found in 90 (80%), 14 (12%), and 8 (7%) children respectively. Epitrochlear node was found in only 1 child. 11 (10%) children had discharging sinus, all being in cervical group. The consistency of nodes was firm in 98 (87%), fluctuation was present in 15 (13%). In 87 (77%) lymph nodes were matted. Hepatomegaly of more than 2 cm was present in 13 (11%) and spleen was enlarged (> 2 cm) in 4 (4%) only: Mantoux test was positive in 96 (85%) children and chest X ray was abnormal in 25 (22%) cases. Findings suggest that tubercular adenitis occurs in all age groups with equal frequency. It can occur in vaccinated children also. It may be a sole manifestation of tubercular infection. The cervical nodes are predominantly involved. There is no typical location of nodes in individual groups but multiplicity and matting of nodes are characteristic features of tubercular adenitis in children. PMID- 10829924 TI - Fulminant hepatitis in Kala-azar. AB - One hundred and fifty cases of Kala-azar were studied for evidence of hepatic involvement. The hepatic function was mildly affected in 25 cases and 3 cases had fulminate hepatitis. Most of the cases were cured after anti-Kala-azar therapy except 2 cases, who died of hepatic failure. This study suggests that fulminant hepatitis may be the outcome of Kala-azar, itself. PMID- 10829926 TI - Effect of adeno-tonsillectomy on hearing threshold and middle ear pressure. AB - Out of 50 children (100 ears) undergoing adeno-tonsillectomy, 34 ears had hearing threshold 20-50 dB (20 dB is normal) and 32 ears showed negative middle ear pressure of 100 to 400 mmH2O (100 mmH2O is normal). Post-operatively only 7 ears had hearing threshold of 20-30 dB and negative middle ear pressure of 100 to 200 mmH2O. Thus adenoidectomy improves eustachian tube functions. PMID- 10829925 TI - Value of antenatal echocardiography in high risk patients to diagnose congenital cardiac defects in fetus. AB - Results of fetal echocardiography in 1062 high risk pregnant patients are described. It was performed before 28 weeks of gestation in 770 cases. These were 38 abnormal scans (3.6%). A fetal arrhythmia was diagnosed in 14 cases and structural abnormality of the heart in 24. Complete atrioventricular block was commonest (n = 12), structural heart disease associated in two of these cases. Other lesions identified were atrioventricular septal defect (n = 5), hypoplastic left heart syndrome (n = 4), ventricular septal defect (n = 4), Ebstein's anomaly (n = 3), coarctation of aorta (n = 2) and others (n = 9). Postnatal confirmatory echocardiography is available in a total of 993 babies including 36 of 38 abnormal cases. There were eleven neonatal deaths amongst babies with abnormal scans. Errors in interpretation were observed in six instances. An anomaly was missed in five cases; in two of these, the main cardiac malformation was picked up but secondary lesions were missed. In one case, a false positive diagnosis of atrial septal defect was made. These errors did not influence the management of the pregnancy. Fetal echocardiography is a very sensitive (91.6%) and specific (99.9%) tool for antenatal diagnosis of congenital heart disease in high risk pregnancies. The information so obtained helps in guiding the optimal obstetric and neonatal management of these cases. PMID- 10829927 TI - Role of antibody dependent cell mediated cytotoxicity in ABO hemolytic disease of the newborn. AB - Study includes fifty O blood group mothers delivering A or B group infants suffering from jaundice and a control group consisting of thirty one O group mothers of non-jaundiced infants. Lytic ability of maternal IgG anti-A/anti-B was determined by 51Cr ADCC assay in which cord blood monocytes were used as effector cells. In control series mean% specific lysis (SL) was 18 +/- 3.1 for IgG anti-A and 17.9 +/- 3.1 for IgG anti-B. In jaundiced series IgG anti-A was more lytic than IgG anti-B. However, the increase in ADCC lysis was statistically insignificant. Even though > 1:32 titre was more often associated with > 35% SL, in general the immune A/B antibody titre showed poor correlation with ADCC lysis. Majority of the severe ABO-HDN cases had > 35% ADCC lysis. PMID- 10829928 TI - Genital standards for south Indian male newborns. AB - One hundred and thirty-five male newborns in JIPMER hospital were studied. The penile length (stretched and unstretched), width as well as testicular length and breadth were measured. The influence of the gestational age and intrauterine growth on these measurements were studied. The mean stretched penile length for term babies was 3.57 cms (+/- 0.46), the unstretched length 3.26 cms (+/- 0.41), and the width was 1.04 cms (+/- 0.15). There was a linear increase in the above measurements with increasing gestational age. The right testicular length was 1.39 cms (+/- 0.28) and breadth was 0.98 cm (+/- 0.15) while the length and breadth for left testis were 1.32 cms (+/- 0.24) and 0.95 cm (+/- 0.14) respectively for term babies. The prepucial type was tubular in 103 (76.3%) babies and of ring type in 32 (23.7%). In 72 (53.3%) babies the prepuce could be retracted enough to visualise external urinary meatus. PMID- 10829929 TI - Empty sella in short children with and without hypothalamic-pituitary abnormalities. AB - A study was conducted on growth hormone (GH) response to oral clonidine (0.15 mg/m2), GH and cortisol responses to i.m. glucagon (0.1 mg/kg), and glucose response to an oral load of glucose (1.75 g/kg). Measurements were made on the circulating concentrations of free thyroxine (FT4), thyroid stimulating hormone (TSH) and different growth parameters and CT sellar images in 25 GH deficient children (Peak GH response to clonidine and glucagon < 7 ug/ml), 15 growth retarded children (Ht < 5th percentile for age and gender) with sickle cell disease (SCD) and GH deficiency, 30 randomly selected children with normal variant short stature (NVSS) (HtSDS 2SD below the mean for age and gender with normal GH response to stimulation (> 10 ug/ml) and 20 age-matched normal children were evaluated. Out of the 25 children with GH deficiency, five had multiple pituitary hormonal deficiency (GH < TSH and/or ACTH. deficiencies), and 20 had isolated GH deficiency. Empty sella, either complete or partial, was detected in 9 out the 20 children with isolated GH deficiency (45%), 4 out of the 5 children with multiple pituitary deficiency (80%), all the children with SCD and GH deficiency (100%), 3 out of the 30 children with NVSS (10%) and in none of the normal children. The insulin-like growth factor-I (IGF-I) concentrations were significantly lower in the two groups of children with GH deficiency compared to those with NVSS. The height standard deviation scores (HTSDS) were significantly lower and the annual growth velocity was slower in children with idiopathic GH deficiency and empty sella compared to those with NVSS and those with empty sella associated with SCD. The bone age delay (yr) did not differ among the 3 groups of children with short stature. All children with isolated GH deficiency associated with empty sella had normal body mass indices (BMI), while all the children with SCD and empty sella had BMI below the 5th percentile for the corresponding age and gender. None of the children had glucose intolerance. In conclusion, children with growth retardation and abnormal hypothalamic pituitary functions have high incidence of empty sella. However, empty sella is detected in considerable number (10%) of short children with normal hypothalamic pituitary function. PMID- 10829930 TI - Effect of gender, birth weight and gestational age on serum 17 hydroxyprogesterone concentration and distribution among neonates in Saudi Arabia. AB - Blood samples from 813 newborns were spotted on to filter paper and the 17 hydroxyprogesterone concentration was measured using the Delfia (R) fluorometric immunoassay. The median, mean, and standard deviation (SD) for the total population were 20, 21 and (11) nmol/L respectively. Males had significantly higher levels than females with median, mean and (SD) of 22, 22 and (12) nmol/L. Similarly, low birth weight babies were found to have significantly higher levels than normal birth weight babies with median, mean and (SD) of 21, 24 (12) nmol/L. Preterm babies were also found to have significantly higher levels than full term babies, with median, mean and (SD) of 25, 29 (16) nmol/L. As experienced elsewhere, those factors should be taken into consideration when considering a cut-off point in any neonatal screening program for congenital adrenal hyperplasia. PMID- 10829931 TI - Mucolipidoses--II: A report of three cases. AB - Mucolipidoses II is a rare lysosomal storage disorder with autosomal recessive inheritance. There cases with typical clinical features in early infancy like coarse facial features, severe psychomotor retardation and joint contractures are being reported. All the cases had no mucopolysacchariduria. These cases had normal values of lysosomal enzymes in leucocytes but markedly increased values in serum thus confirming mucolipidoses II. Despite the fact that there is no specific treatment, genetic counselling and prenatal diagnosis is indicated. PMID- 10829932 TI - Partial trisomy of chromosome 10 inherited from a carrier father. AB - Partial trisomy of chromosome 10q is a very rare condition with only four cases having been reported int he literature. This report describes a neonate with symmetric growth retardation and multiple dysmorphic features, in whom G-banded chromosomal analysis revealed a partial trisomy of chromosome 10q (q2.4-q ter). The father was diagnosed as a carrier of a balanced translocation with a karyotype of 46, XY t(10.3) (q2.4L : pter). In patients with a bad obstetric history, genetic counselling prior to a new conception cna aid in early prenatal diagnosis of fetuses with recurrent chromosomal abnormalities by means of fetal tissue sampling. PMID- 10829933 TI - Fatal pulmonary infection caused by Mycobacterium smegmetis in an infant. AB - Mycobacterium smegmetis is a rapidly growing mycobacteria which is usually regarded as an environmental saorophyte. Recently it has been isolated from human beings mainly from skin and soft tissue lesions. Pulmonary lesions are extremely rare. A case of pulmonary consolidation caused by M. smegmetis in an infant is reported. The outcome was fatal. The organism was resistant to various antitubercular drugs. A brief review is presented. PMID- 10829934 TI - Splenic abscess following blunt abdominal trauma. AB - A rare case of splenic abscess occurring after blunt abdominal trauma in a previously healthy boy is reported. The diagnosis was made by ultrasonography. The patient recovered after splenectomy and drainage of subphrenic and intraperitoneal pus. PMID- 10829935 TI - Pharyngeal palsy in enteric fever. PMID- 10829937 TI - Hoarseness in a child--an unusual cause. PMID- 10829936 TI - The enigma of Indian plague--the U.A.E. experience. PMID- 10829938 TI - When should pediatrician reach the delivery room? PMID- 10829939 TI - Epidemiology of childhood hepatitis B in India: vaccination related issues. AB - It has been estimated that presently hepatitis B kills more people every day than AIDS kills in a year world-wide. Infection with hepatitis B produces a wide range of manifestations ranging from asymptomatic carriers to persistent infections leading to chronic liver diseases and hepatocellular carcinoma. Availability of effective and safe vaccine has made all this preventable. To formulate on appropriate vaccination strategy for India the epidemiology of hepatitis B needs to be defined. This report critically reviews the available data. The burden of long term sequelae of HBV infection is probably under-diagnosed and under reported in India. Prevalence studies of HBV markers indicate that India falls under the area of intermediate endemicity. Limited data on age-specific prevalence of HBV markers suggests that the majority of the infection seems to take place below 15 years of age, and most of it under one year. Perinatal transmission appears to contribute significantly to the carrier pool. Childhood vaccination for HB among the general population is the obvious strategy of choice. But more information is required to decide on the timing of the first dose. PMID- 10829941 TI - HBV control in India. PMID- 10829943 TI - The art and science of pediatric diagnosis. PMID- 10829942 TI - Chronic viral hepatitis. AB - Among hepatitis A to E viruses, hepatitis B, C, and D viruses can cause chronic hepatitis, in both children and adults. Hepatitis B virus (HBV) infection is the most prevalent and important one. Perinatal transmission accounts for about 40 45% of chronic HBV infection in hyperendemic areas. Horizontal transmission through intramuscular injection using non-sterile needles and intrafamilial spread accounts for the other half of carriers. During the natural course of HBV infection, the host gradually clears HBV and hepatitis B e antigen (HBeAg), liver damage and elevation of aminotransferases occur during the process of HBV clearance. The most effective way to eliminate HBV infection is immunoprophylaxis starting since birth. It can prevent both HBV and hepatitis D virus (HDV) infections. Hepatitis C virus (HCV) infection in children occurs mainly in high risk children, such as those who received blood product or injection using non sterile needles, or infants of HCV viremic mothers, etc. Screening of blood product reduced markedly the prevalence of post-transfusion HCV infection, but the prevention of sporadic cases requires HCV vaccination which is still under investigation. PMID- 10829940 TI - Hepatitis B, C & D viral markers in multitransfused thalassemic children: long term complications and present management. PMID- 10829944 TI - Breast is best. PMID- 10829945 TI - Delta-hepatitis. AB - Investigations were conducted for serological evidence of hepatitis B virus (HBV) and hepatitis D virus (HDV) infections in children suffering from acute viral hepatitis. A total of 52 serum samples were analysed by enzyme immunoassay. Of these, 18 (24%) were positive for hepatitis B virus markers and 34 (65.4%) were negative. Delta virus infection was detected in 6/18 (33%) hepatitis B patients. A significant finding was, that of the 34 patients negative for hepatitis B, 4 (12%) were positive only for HDV although the latter can only occur as a coexistent infection with hepatitis B virus. From the present study it may be inferred that delta virus infection is prevalent in children and absence of HBV markers does not rule out hepatitis D. PMID- 10829946 TI - Paradoxical response to antitubercular drugs. AB - Seven patients with paradoxical response to antitubercular drugs are reported. In three cases of intracranial tuberculomas, newer lesions appeared and in two cases preexisting tuberculomas enlarged. In two cases of tubercular meningitis, multiple tuberculomas appeared. All these cases exhibited newer symptoms and CT/MBI revealed the paradoxical response to antitubercular drugs. All responded to continued conservative therapy, with addition of pyrazinamide. PMID- 10829947 TI - Multiple drug resistant non typhoidal Salmonella spp associated with acute diarrheal disease. AB - Rectal swabs/stool specimens from 115 children (0-5 years) suffering with acute diarrhea were screened for non typhoidal salmonella species. 7 (6%) patients were found to be positive for non typhoidal salmonella. 4 (3.47%) were positive for S. paratyphi B and 3 (2.6%) were positive for S. typhimurium. Multidrug resistance was seen in 57 percent of the strains. All strains were sensitive to Ciprofloxacin. All strains were resistant to Ampicillin followed by Ciprofloxacin. All strains were resistant to Ampicillin followed by Gentamycin (43%), Kanamycin (43%), Tetracycline (43%), Streptomycin (28.5%) and Chloramphenicol (28.5%). PMID- 10829948 TI - Infant feeding--knowledge and attitudes in a rural area of Karnataka. AB - A study of the knowledge and attitude towards infant feeding was conducted in South India on a group of 300 mothers whose babies' ages from 3 days to 17 months. Most of the mothers opined that breast-feeding had to be initiated within 24 hours of birth. However, only 32% felt that breast milk should be the first feed, whereas 68% considered prelacteal feeds a necessity. 71% of the mothers considered 3 to 5 months to be the optimum duration of exclusive breast-feeding. 90% felt that cow's milk was an ideal supplement. 78.3% subscribed to the view that breast-feeding should continue beyond one year. A sizeable proportion expressed the need to discontinue breastfeeds during babies' or maternal illness, particularly diarrhoea. Our results will form the babies of the content and the target groups in our future health education programme. PMID- 10829949 TI - Urinalysis in urinary tract infection. AB - In 376 children, between 6 months and 5 years of age, with suspected urinary tract infection, 4 parameters of a routine urine examination were correlated with culture reports. In diagnosing urinary infection, the sensitivity and specificity of proteinuria was 79 and 80% respectively, that of bacteriuria 78 and 96% and that of pyuria > 10 wbc/hpf 80 and 82% respectively. 61% among the culture positive groups had all these three parameter present, as against only 0.5% in the culture negative group (P < 0.001). All these 3 parameters were absent in 70% in the culture negative group, as against 8% in the culture positive group (P < 0.001). Bacteriuria in association with either proteinuria or Pyuria > 10 Wbc/hpf had 98% specialty is diagnosis. In diagnosis UTI, Pyuria > 10 wbc/hpf was significantly more specific (82:66.6) than the conventional > 5 wbc/hpf. Isolated proteinuria, isolated pyuria, isolated bacteriuria and microscopic haematuria were not features of urinary tract infection in children. Urine culture can be more selectively done if the routine urinalysis is well interpreted. In the absence of dependable culture facilities, a routine urine examination can be reliably used in the diagnosis of UTI in children. PMID- 10829950 TI - Reversibility of mitral regurgitation following rheumatic fever: clinical profile and echocardiographic evaluation. AB - The clinical disappearance of the murmur of rheumatic mitral regurgitation after period of time has been documented by many researchers. However no studies have related the disappearance of the murmur with the functional or anatomical state of the mitral valve. This study was done to elucidate the mitral valve status using doppler and color coded echocardiography among those children who have lost their apical pansystolic murmur on auscultation following a documented attack of rheumatic fever. The study sample consisted of 51 patients including 31 patients in whom the murmur has disappeared (group I), and 20 patients with persistent isolated mitral regurgitation (group II). Patients of group I had significantly lower grades of murmur intensity, lower incidence of cardiomegaly, and had no heart failure in the initial attack. They were more compliant with prophylaxis and had less recurrences than patients of group II. The murmur disappeared in patients of group I from 1/2 to 14 years after the initial attack. Echocardiography revealed that such patients had a normal mitral valve apparatus, and a normal heart size and function. Only 5 patients of this group had a significant regurgitant jet demonstrated by colour doppler. We concluded that recovery of the mitral valve and return of cardiac functions to normal is possible in patients who had mitral regurgitation following rheumatic fever. Some of them may still have an inaudible mild regurgitation. Patients who have lost their murmur may be allowed to exercise freely, yet penicillin prophylaxis should not be discontinued. PMID- 10829952 TI - High risk register--an economical tool for early identification of hearing loss. AB - Deafness needs to be dignified nearly as its adverse effects increase with the degree and age of onset. The high risk register helps to alert the professional to suspect the presence of this hidden handicap. Histories of 281 hearing impaired children and 158 normal children was collected and analysed. Risk items that appeared significant were--rashes with fever during pregnancy, family history of childhood deafness, asphyxia at birth or blue baby, prematurity (less than 32-34 gestational weeks), low birth weight (less than 1200 gm), hyperbilirubinemia, birth defects of ear nose, throat and head, consanguineous marriage (cousin), parental concerns about their child's hearing. These high risk factors could help in identifying hearing impairment early and in envisaging quality habilitation programs. PMID- 10829951 TI - Epidemiology of chronic suppurative otitis media and deafness in a rural area and developing an intervention strategy. AB - Of 613 children evaluated in a village in Haryana 94 (15.3%) were observed to have chronic suppurative otitis media (CSOM). Fifty eight (61.7%) children had hearing impairment. CSOM contributed to 71.6% of the hearing impaired (58/81). On analysis of association of CSOM with literacy and socio-economic status of mothers, and age, sex, and upper respiratory tract infections (URI) in children positive correlation was observed only with URIs (P < 0.001). Literacy and socio economic status of the mothers did not correlate significantly with knowledge about treatment seeking, and ear cleaning practices, probably due to the narrow range of incomes and literacy levels. An intervention program consisting of play, demonstrations, health charts and slogans, and aural cleaning and antibiotic drops was introduced. PMID- 10829953 TI - Coping and adaptation in parents of children suffering from acute lymphoblastic leukemia. AB - The coping and adaptation of 30 parents of children suffering from A.L.L. (during first remission), was studied with the Thematic Apperception Test (TAT--Indian modification). Emotional distress was evident in the stories of 83%, only 37% maintained an expectation of a positive outcome. Parents' psychiatric morbidity was found to be associated with the gender of the parent, employment status, and on the TAT--with the expectation of an unfavourable outcome, feelings of lack of self efficacy and emotional distress. Particularly depression. A significant association between the presence of psychiatric disturbance in the children and the parents was also found. PMID- 10829954 TI - Henoch-Schonlein purpura with thoraco-lumbar spine involvement precipitated by trampoline jumping. AB - A nine-year-old boy with Henoch-Schonlein Purpura (HSP) presented with an arthropathy of the thoraco-lumbar spine. The boy played on a trampoline for several days before the onset of the low-back pain. The vascultic rash of HSP is recognized to be pressure-dependent. The authors speculate that the arthropathy of HSP is also pressure-dependent and that the low back pain in this patient was consequent to the preceding trampoline jumping. PMID- 10829955 TI - Pelvic lipomatosis. PMID- 10829956 TI - Acute quadriplegia as a presenting symptom of acute myeloid leukemia. PMID- 10829957 TI - Giant cystic hygroma of pelvis in a neonate. PMID- 10829958 TI - Pulse Polio Program--a national perspective. PMID- 10829959 TI - Oral rehydration fluids for use at home. PMID- 10829961 TI - Pulmonary disorders and their management. PMID- 10829960 TI - Sedation in pediatric patients. AB - Sedation is being used increasingly in children to allay anxiety and discomfort. Sedation can also increase the efficiency of performing both diagnostic and therapeutic procedures in children. There are a wide array of available sedation methods that are used by radiologists, gastroenterologists, hematologists/oncologists and emergency room physicians everyday. Indiscriminate use of sedatives has led to seizures, respiratory arrests and death in a variety of practice settings. With improved monitoring capability, more potent drugs and better understanding of the pharmacokinetics in children, it is possible to provide batter care. PMID- 10829962 TI - Childhood asthma--advances in pathogenesis. AB - Increase in morbidity and mortality of asthmatics in the world is a cause of concern. Many researchers have described various aspects of etiopathogenesis which has thrown light on the better understanding of asthma. Our experience with nearly 3 lakhs of asthmatic children, over a period of twenty-five years and our studies in Asthma clinic of ICH & HC, Madras generated new ideas to propose a hypothesis on etiopathogenesis of asthma. "Asthma is a disease caused by a specific infective agent in a genetically predisposed individual resulting in altered cellular response initially leading to hyperactive bronchial tree which on exposure to various aggravating factors manifest clinically as recurrent cough, dyspnoea and wheeze". Category of wheezers who manifest asthma is also discussed. PMID- 10829963 TI - Childhood asthma--office management. AB - Childhood asthma is a major problem in office practice. For an acute life threatening attack (which is indicated by presence of severe distress, pulsus paradoxus, oxygen saturation less than 93%, cyanosis, peak expiratory flow rate of less than 50% of predicted) child should be directly admitted in intensive care unit. If it is a non life threatening attack the child can be managed in the office. Initially, the child should be given b2 agonists by inhalation route with either metered dose inhaler (MDI), MDI with spacer or nebuliser. It there is severe bronchospasm or inhalation therapy is not possible then epinephrine of b2 agonists may be given subcutaneously. The medications can be repeated 2-3 times. If response is adequate the child may be sent home on b2 agonist by oral or inhalation route at an interval of 406 hours. In case of inadequate response the child is started on oral or parenteral corticosteroids. Even after steroids if inadequate response the child is started on intravenous theophylline. Once the acute exacerbation is controlled the child is assessed for starting maintenance therapy. For this purpose his illness is graded from stage I to V depending on the severity. For stage I and II b2 agonists are prescribed as and when required. For stage III sodium cromoglycate by inhalation should be prescribed. For stage IV inhalation steroids in usual doses and for stage V inhalation steroids in higher doses are prescribed along with a minimum dose of oral steroids is added. For symptomatic control slow release theophylline or long acting b2 agonists may be added along with maintenance therapy as and when required. Apart from medications a proper education of parents and patients is necessary to improve the outcome of asthma by increasing the compliance and better control of environment. PMID- 10829964 TI - Acute bronchiolitis--recent advances in treatment. PMID- 10829965 TI - Management of respiratory failure. PMID- 10829966 TI - Dexamethasone therapy in chronic lung disease. AB - Steroids have been used in the treatment of infants with chronic lung disease (CLD) for over a decade. Some studies have reported beneficial effects from long, tapering 42-day course of dexamethasone. Short term regimens have also shown beneficial effects on ventilator dependent infants with CLD. Although steroid therapy has been successful in infants with established CLD, more recently, dexamethasone therapy is being initiated in infants with RDS considered to be at risk for developing CLD. Some of the initial studies reported higher rates of infection, but more recent prospective data have not shown an increased incidence of sepsis in patients treated with steroids. Presently, early steroid therapy appears to be beneficial to minimize lung injury in infants treated with surfactant. PMID- 10829967 TI - Exponential model for weight using 'RJ index'. AB - We screened a sample of 204 children between the age of 3 months to 5 years and 3 months were during a general health survey conducted by the Department of Community Medicine, Kasturba Medical College, Mangalore from March 1992 to December 1993 at Kumpala village under PHC Ullal about 10 kms from Mangalore City. We developed a new index RJ as RJ = Mid upper Arm Circumference x (Height)2 x 10(-4) and tested the correlation of RJ index and weight for age. We found the correlation coefficient (r = 0.8) to be very significant (P < 0.001). Thus we propose the model as Weight = A x exp (RJ x B). We found that the available data fit in this model. We hope that this study will be a new step in looking at arm circumference as a tool in estimating weight of pre-school children. PMID- 10829969 TI - Microbial contamination of weaning foods. AB - A survey was conducted to determine the prevalence rate of Escherichia coli in different foods being fed to children in the weaning age group. The samples were collected from middle income group (MIG) and high income group (HIG) houses in different localities in Chandigarh. A total of 530 MIG and 525 HIG houses were selected on the basis of availability of children in the weaning age group. A total of 2,016 samples of weaning foods were collected. Isolation of Escherichia coli from MIG and HIG houses was 66.75% and 8.5% respectively. Enterotoxigenic Escherichia coli were detected in 61.47% and 43.5% of the total Escherichia coli isolates from MIG and HIG houses respectively. 'O' serogrouping of the isolates was done and the predominant serotypes found were 06, 0106, 043, 057, 01. The laboratory studies showed that almost all weaning food samples prepared in the laboratory supported the growth of known enterotoxigenic Escherichia coli. PMID- 10829968 TI - C-reactive protein in childhood meningitides. AB - Utility of C-reactive protein (CRP) latex agglutination test in meningitis was evaluated. Serum CRP test was positive in 100% cases of meningitic groups and 53% cases of "no meningitis (NM)" group. Cerebrospinal fluid (CSF) CRP test was positive in 100% cases of pyogenic meningitis, whereas it was negative in 95% cases of tuberculous meningitis and 100% cases of NM group. CSF CRP test showed 100% sensitivity and negative predictive values, 95-100% specificity and 94-100% positive predictive values for various inter-group differentiations. This study concluded that CSF CRP positive cases should be considered as pyogenic meningitis unless proved otherwise. Routine use of this simple, reliable and inexpensive test is recommended for rapid diagnosis and differential diagnosis f meningitis. PMID- 10829970 TI - Manual and pump methods of expression of breast milk. AB - This study was designed to compare two methods of breast milk expression, namely, the manual and the pumping method using a hand-held cylindric pump. The parameters evaluated were (i) the output of breast milk during milk expression sessions of 15 minutes' duration, and, (ii) the subjective preference of the method by the mothers. In the first phase, 22 mothers whose infants were on gavage feeding in the nursery, had 3 sittings each by the two methods on 4th and 5th postnatal days (66 expression). It was seen that the use of breast pump (Medela) was associated with significantly higher volume of breast milk expressed per session (41.57 +/- 16.05 ml vs. 21.7 +/- 10.5 ml, P < 0.001). In the second phase, 14 mothers had such sessions (42 each) not only on the 4th and 5th postnatal days, but lso on days 8 and 9. It was again seen that, the volume of breast milk expressed was greater by the pump method than the manual expression (on day 5 and 6 (46.8 +/- 26.3 ml vs 31.2 +/- 15.5 ml, P < 0.01) as well as on day 8 and 9 (50.40 +/- 11.2 ml vs 38.49 +/- 13.4 ml, P < 0.01). Subjectively, the pump expression was preferred by the mothers on day 4 & 5, while the manual expression was the preference on days 8 & 9. The use of breast pump is more efficient than the manual system of expression of breast milk among mothers whose infants are not directly breast-fed. It is recommended that in case the mothers prefer to use the manual method, let them express as much milk as possible by this method initially, and then follow it up with a short period of pumping to ensure complete evacuation of breasts. PMID- 10829971 TI - Epidemiology of respiratory distress of newborns. AB - The present prospective study was conducted to find out the incidence, etiology and outcome of respiratory distress (RD) in newborns. All newborns (n = 4505), delivered at this hospital over a period of 13 months, were observed for respiratory problems. Relevant antenatal, intranatal and neonatal information was noted. Cases were investigated for the cause of respiratory distress and followed up for the outcome. The overall incidence of RD was 6.7%. Preterm babies had the highest incidence (30.0%) followed by post-term (20.9%) and term babies (4.2%). Transient tachypnea of newborn (TTN) was found to be the commonest (42.7%) cause of RD followed by infection (17.0%), meconium aspiration syndrome (10.7%), hyaline membrane disease (9.3%) and birth asphyxia (3.3%). TTN was found to be common among both term and preterm babies. While Hyaline membrane disease (HMD) was seen mostly among preterms, and Meconium aspiration syndrome (MAS) among term and post-term babies. Overall case fatality ration for RD was found to be 19%, being highest for HMD (57.1%), followed by MAS (21.8%) and infection (15.6%). Our results indicate that RD is a common neonatal problem. TTN accounts for a large proportion of these cases. MAS and infection also contribute significantly and are largely preventable. Without adequate ventilatory support HMD and MAS carry high mortality. PMID- 10829972 TI - Awareness of pulse polio immunisation. AB - Mass polio immunisation campaign was launched in the national capital territory of Delhi with 2 doses of polio vaccine to be administered to children upto 3 years of age on October and December 4, 1994 respectively. Massive information, education & communication (IEC) efforts through mass media and interpersonal communication preceded the dates of the campaign. A study to assess the awareness of general population was carried out by interviewing 225 adult residents of Delhi using a structured questionnaire. These were drawn by two stage stratified random sampling. Zonewise assembly segments in the first stage and census enumeration blocks in the second stage formed the sampling frame. The study, carried out 3 days prior to date of administration of first dose of oral polio, revealed that 60.4% of population was aware of the programme being launched and 31.6% about aim of the programme. None of the respondents were aware of all the specific parameters put together correctly viz., objective, immunisation days, age group & immunisation status of children. The higher level of awareness was directly proportional to the level of education. The overwhelming success of the programme was indicated by immunisation of > 90% children upto 3 years of age all over Delhi in the first phase of the programme. The key to success of the programme despite low awareness is explained on the basis of unflinching efforts put in by vaccine centre level committees, integrated child development scheme (ICDS) and urban basic service (UBS) functionaries in mobilising people to reach various vaccination centres. Other states planning to launch such mass campaigns should pay attention to social mobilisation in addition to IEC efforts for successful completion of the programme. PMID- 10829973 TI - Iodine deficiency disorders in Bangladesh. AB - An extensive iodine deficiency disorders survey was conducted in Bangladesh in 1993 to assess the latest iodine nutriture status of the country. The clinical variables of the survey were goitre and cretinism, and the biochemical variable was urinary iodine. The "EPI-30 cluster" sampling methodology was followed for selecting the survey sites. In each survey site, the study population consisted of boys and girls, aged 5-11 years, and men and women, aged 15-44 years, in about equal populations. The total number of survey sites was 78 and the total number of respondents was 30,072. The total number of urine samples was 4512 (15% sub sample). The current total goitre rate (grade 1 + grade 2) in Bangladesh is 47.1% (hilly, 44.4%; flood-prone, 50.7%; and plains, 45.6%). The prevalence of cretinism in the country is 0.5% (hilly, 0.8%; flood-prone, 0.5%; and plains, 0.3%). Nearly 69% of Bangladeshi population have biochemical iodine deficiency (urinary iodine excretion [UIE] < 10 mg/dl) (hilly, 84.4; flood-prone, 67.1%; and plains 60.4%). Women and children are more affected that men, in terms of both goitre prevalence and UIE. The widespread severe iodine deficiency in all ecological zones indicates that the country as a whole is an iodine-deficient region. Important recommendations of global interest are made from the experience of the survey. PMID- 10829974 TI - Benign pleomorphic adenoma of sub-mandibular gland. PMID- 10829975 TI - Transient hyperammonemia of newborn. PMID- 10829976 TI - Mental retardation, short stature and brittle hair (BIDS syndrome; hair brain syndrome). PMID- 10829978 TI - Metaphyseal chondrodysplasia--a differential diagnosis of rickets. PMID- 10829977 TI - Optimal care for children with Down syndrome in India. AB - Medical problems which arise in Down Syndrome are reviewed. Guidelines for management including early diagnosis of congenital heart defects, gastrointestinal anomalies, eye problems, hearing defects, thyroid dysfunction, at lanto-axial instability, prognosis for mental retardation, and role of early stimulation are provided. Prenatal diagnosis in high risk pregnancies is recommended. PMID- 10829979 TI - Evaluation of mass pulse immunization with oral polio vaccine in Delhi: is pre registration of children necessary? AB - Delhi was the fourth State in India to conduct mass immunization of children (Pulse Polio Immunization) of the < 3 year age group with Oral Polio Vaccine (OPV) as a strategy towards the eradication of poliomyelitis. This study attempted to evaluate the immunization coverage achieved and the channels of communication which were effective in increasing coverage in three high risk areas of Delhi during October 1994. The overall immunization coverage was 89%. Information sources like enumeration visits, posters, television, radio and schools statistically correlated with the Pulse Polio Immunization (PPI) outcome. However, the cost of enumeration was high. Other less expensive channels of communication appeared to be equally effective. Only 11% of the children surveyed were not immunized with PPI OPV. The major reasons why some children did not receive OPV was that parents were "not informed" or they were "too busy". PMID- 10829980 TI - Impact of passive smoking. PMID- 10829982 TI - Pulmonary function tests. AB - Spirometry is indicated in all the children with clinical diagnosis of asthma, chronic/recurrent cough or wheeze, exercise induced cough or breathlessness and with recurrent respiratory manifestations. Mid expiratory How 25-75% (MEF 25-75) is an important diagnostic parameters in children due to its effort independence, high sensitivity in bronchodilator reversibility test and also because it represents smaller airways and is likely to be affected in mildest obstruction. The base line spirometry, bronchodilator reversibility and histamine challenge are diagnostic of hyper reactive airway with 98% sensitivity. Flow volume loops hardly add to the diagnosis and also need more co-operation from the subject. Therefore it is not useful in children. PEFR monitoring constitutes an important part of the followup care of asthma patients. PMID- 10829981 TI - Characterization of the biochemical abnormality in the Smith-Lemli-Opitz syndrome. PMID- 10829983 TI - Advances in diagnosis of tuberculosis. AB - The diagnosis of childhood tuberculosis is acknowledged to be an imprecise process since bacteriological confirmation is available in only 30-40% of cases. Newer developments in diagnosis of tuberculosis include use of fluorescent stains for smears, newer systems for radiometric detection of mycobacteria, rapid sensitivity testing using firefly bioluminescence, liquid chromatographic analysis of mycolic acids, immunodiagnostics for M. tuberculosis specific antigens and the impact of molecular diagnostics with amplification methods. The search for simple, reliable test for early stages of the disease (in particular TB meningitis) still continues. PMID- 10829984 TI - Multi drug resistant tuberculosis. PMID- 10829985 TI - Bronchoalveolar lavage in pediatrics. AB - Bronchoalveolar lavage is a relatively new technique that is used to study the local cellular, biochemical and immunological changes occurring in the lower respiratory tract. The procedure involves instilling a fixed volume of saline into a lung segment after the flexible fibreoptic bronchoscope is wedged into a distal bronchus. The saline is aspirated back and can be used for microbiological and other studies. Recently, attempts have been made to standardise the procedure in children and obtain data on BAL cellular profile in healthy children. The main indications for BAL are diagnostic, particularly to diagnose unusual infections in immunocompromised children. It is also helpful in the diagnosis of a number on non infectious conditions, based on the cellular profile and other constituents. With the availability of new techniques like flow cytometry, analysis of lymphocyte and other cell subsets has become possible leading to a better understanding of the immunopathogenesis of various respiratory diseases. PMID- 10829986 TI - Flexible fiberoptic bronchoscopy. AB - The value of flexible fiberoptic bronchoscopy in children are increasing day by day, but still underutilized even in many advanced institutions. 630 Fiberoptic bronchoscopy procedures under local anaesthesia were performed for various clinical conditions for diagnosis and therapy. Nasal route is preferred than oral route. Therapeutic indications are more in children than adults. Nearly one third of children with collapse, consolidation have shown radiological clearance after repeated bronchoscopy. PMID- 10829987 TI - Practical approach to recurrent respiratory infections. AB - Respiratory diseases are a major cause of morbidity and mortality in developing countries. Recurrent respiratory infections in children pose a great challenge to the pediatrician where he has to exercise his clinical acumen and methodical approach for correct diagnosis and treatment. It is a fact that children should suffer 7 to 8 upper respiratory infections per year until they are 5 years of age when their immune status reaches adult level. In this situation, it is essential to find out whether the frequencies are abnormal. Whenever a child has the following problems, then only it needs to be investigated.--(a) repeated bacterial pneumonias; (b) a child less than 3 months old having repeated respiratory infections; (c) a child of 9 months old without a history of exposure infections; (d) infections complicating into bronchiectasis and; (e) in a child where there is no history of allergy or asthma. Once the problem is established as a true recurrent respiratory infection, the clinician should pose questions- whether it is chronic, acute or recurrent, to find out the site of pathology, seriousness of the problem, response to previous medications, to establish the possible diagnosis which fall into six categories--congenital anamolies, aspiration syndrome, genital disorders, immunological diseases, immune deficiency disorders and allergic diseases. The author discusses quoting some examples for various categories avoiding non pulmonary causes for recurrent respiratory infections in children. PMID- 10829988 TI - Cystic fibrosis--an Indian perspective on recent advances in diagnosis and management. AB - Cystic fibrosis (CF) is a common inherited disorder in caucasians. The estimated incidence of CF in Asians varies from 1:10,000 to 1:12,000. Indian data is restricted to few case reports. The gene for CF is located on the long arm of chromosome 7 at position 7q13. There are more than 300 identified mutations in CF. The basic defect in CF is a mutational change in the gene for chloride conductance channel. Failure of chloride conductance by epithelial cells leads to dehydration of secretions that are too viscid and difficult to clear. The disease is characterized by abnormal secretions in the respiratory, gastrointestinal and reproductive tract and sweat glands. The common clinical manifestations include meconium ileus in neonatal period, recurrent lower respiratory tract infections (pseudomonas pneumonia, bronchiectasis), steatorrhoea, azoospermia, and in late stages hepatobiliary and endocrine pancreatic dysfunctions. The diagnosis of disease is established by clinical criteria and sweat chloride concentration more than 60 mEq/L. Facilities for DNA diagnosis of common CF mutations are now available in India. The treatment of CF includes early diagnosis, daily clearance of respiratory passages, appropriate antibiotic therapy, aerosolised recombinant human DNase and antibiotics, and nutritional supplementation. The latter include changes in diet composition, pancreatic enzyme supplementation and vitamins and trace mineral supplementation. Gene therapy for the pulmonary manifestations is being tried in a number of centres abroad. Other considerations include heart lung transplantation and ameloride inhalation therapy. PMID- 10829989 TI - Serum, zinc and copper levels in children with protein energy malnutrition. AB - Serum zinc and copper were measured by atomic absorption spectrophotometry in 58 children (3 months-5 years); of these, 46 children had protein energy malnutrition (PEM), and 12 children served as controls. The levels of serum zinc and copper were found to be significantly low in children with severe malnutrition (grades III and IV PEM). There was a significant positive correlation between serum zinc and height-for-age (r = 0.8809, p < 0.001). Serum copper was found low only in children exhibiting marked linear growth retardation (height-for-age < 85% of the normal). Hypoalbuminemia (serum albumin < 2.5 g/dl), and anemia (hemoglobin < or = 8.0 g/dl) in malnourished children were associated with significant decline in serum zinc and copper levels, respectively. PMID- 10829990 TI - Infective endocarditis in infants and children. AB - Due to changing characteristics of infective endocarditis in the past two decades, we, retrospectively analysed 28 cases of infective endocarditis in children of age less than 15 years at Sher-i-Kashmir Institute of Medical Sciences, Soura, Srinagar from December, 1983 to November, 1993. The incidence of disease was observed as 1.5 cases/1000 children admitted with a M:F ratio of 2:1. Three patients were of age less than 2 years (group I) as 25 were above 2 years of age (group II). The two groups had significant difference in portal of entry of infection, infective microorganisms, echocardiography and prognosis. Congenital heart disease was the commonest underlying cardiac lesion in 24 (85.71%) patients. Portal of entry of infection was apparent in 35.71% only; dental route being more frequent in group II. Streptococcus viridans (in 9 cases) followed by staphylococcus aureus (in 4 cases) were the two common organisms isolated. Patients were treated, for a period of 4-6 weeks with a over all mortality rate of 25%. Factors associated with poor prognosis were age < 2 years, staphylococcal infection ad negative blood cultures. Heart failure resistant to medical therapy was a leading cause of death. PMID- 10829991 TI - Auditory profile in children recovering from bacterial meningitis. AB - In the present study BERA profile of 30 post-meningitic children was compared with 15 normal children of the same age and it was observed that 36.6% children in the age range of 6 months to 36 months were found to have varying degree of sensorineural deafness. Severe bilateral sensorineural hearing loss (> 80 dB) was observed in 6.6% children and moderate (40-80 dB) hearing loss in 30% of children. Abnormalities were bilateral in both the samples of children with severe hearing loss (> 80 dB) whereas among 9 children who had moderate hearing loss abnormalities were bilateral in one patient and unilateral in the remaining 8 children. A relationship between higher incidence of sensorineural deafness and younger age of children, and occurrence of seizures during meningitis were noted. But no relationship was observed with either sex, hydrocephalus, subdural effusion or with low CSF sugar and high CSF proteins. PMID- 10829992 TI - Brainstem auditory evoked response (BAER) in childhood bacterial meningitis. AB - Brainstem auditory responses were recorded in 50 children of bacterial meningitis and age matched 50 normal children. Abnormal BAER was found in 32 (64%) patients of bacterial meningitis. These abnormalities included prolonged latency (56.2%); unilateral absent response (25%); bilateral absent response (25%) and prolonged interwave interval (25%). Follow-up could be done in 23 patients of 46 survivors. All the patients with prolonged latency either became normal or improved. In majority of the patients having absent response, the abnormality persisted. Abnormal BAER was significantly associated with age < 2 years (p < 0.02), Modified GCS Score < or = 8 (p < 0.001), Seizures (p < 0.02), raised Intracranial Pressure (ICP) (p < 0.02) and CSF sugar < 20 mg% (p < 0.05). PMID- 10829993 TI - Neonatal thrombocytopenia due to pregnancy induced hypertension. PMID- 10829994 TI - Meconium peritonitis--a leading cause of neonatal peritonitis in Kashmir. AB - Meconium peritonitis is a chemical reaction of peritonium to meconium which occurs due to leakage of meconium into peritoneal cavity as a result of perforation of intestines antinatally which gets subsequently sealed. In the present retrospective study, 39 cases of neonatal peritonitis were studied. Meconium peritonitis was diagnosed if a) abdominal X-ray showed diffuse calcifications (b) abdominal paracentesis showed meconium aspirate and c) leprotomy examination. Twenty (51.3%) out of 39 cases of neonatal peritonitis were found to have meconium peritonitis. These included 14 boys and 6 girls, birth weight ranged from 1500 gms to 3200 gms and mean age of presentation included abdominal distension in 100% cases, H/O not passing meconium in 50% cases, Ascites in 45% cases and vomiting in 40% cases, 30% presented with abdominal mass. Pseudocyst formation on X-ray was seen in 23% cases which was much higher than reported in world literature. Overall mortality in meconium peritonitis was 80%. The incidence of meconium peritonitis in this part of country in much higher than reported in Western and Indian literature. The high incidence may be due to underlying cystic fibrosis because Kashmir has a more homogenous population and consanguinity is very common. A prospective study including sweat chloride testing needs to be undertaken to look into the cause for proportionately higher occurrence of meconium peritonitis in Kashmir with particular reference to cystic fibrosis. PMID- 10829995 TI - Newer anti-epileptic drugs. AB - During the past few years, a number of drugs have been added to the anti epileptic arsenal. This review focusses on five of these drugs which have undergone extensive trials: Vigabatrin, Lamotrigine, Gabapentin, Felbamate and Oxcarbazepine. Some of these antiepileptic drugs appear to be helpful for treatment of catastrophic childhood epilepsies. Vigabatrin appears promising in children with infantile spasms who do not respond to ACTH or Prednisolone. Children with Lennox-Gastaut syndrome may respond to treatment with Lamotrigine or Vigabatrin. Gabapentin and vigabatrin have proved to be effective in refractory partial seizures. Oxcarbazepine, a ketoderivative of carbamazepine, is as effective as Carbamazepine but has a better safety profile. Lesser neurotoxicity and fewer drug interactions is another advantage with these drugs. However monitoring is required to determine the long term safety with their usage. These drugs have a definite role in childhood epilepsies refractory to conventional antiepileptic drugs. PMID- 10829997 TI - Congenital chloride diarrhoea. PMID- 10829996 TI - Vitamin E updated. AB - An improved understanding of its physiology has led to better therapeutic use of vitamin E in recent years. It is a physiological membrane bound antioxidant, protecting cell membrane lipids from oxidant damage by free redicals. Cholestatic liver disease, abetalipoproteinemia and ataxia with vitamin E deficiency are the common deficiency states where vitamin E is of definite therapeutic value, while reports of unproven benefits abound in literature. Vitamin E status of the body can be assessed by serum levels and various functional studies. The new water soluble form, tocopherol polyethylene glycol succinate (TPGS), is therapeutically superior to the standard oral forms available. Details of physiology and therapeutic application of the vitamin are discussed. PMID- 10829998 TI - Recurrent trichobezoar: first reported case. PMID- 10829999 TI - Glycogen storage disease type IIa. PMID- 10830000 TI - Abetalipoproteinemia in an Indian family. PMID- 10830001 TI - Large bowel tuberculosis presenting with bleeding per rectum. PMID- 10830002 TI - Juvenile rheumatoid arthritis--clinical manifestations. AB - Chronic inflammatory arthritis in childhood could be due to an obvious cause (e.g. sepsis, rheumatic fever, systemic lupus erythematosus etc.), or it could be idiopathic. After excluding those with obvious cause there still remains a large group of chronic inflammatory arthritis in childhood. This category has been variously called 'juvenile rheumatoid arthritis', 'juvenile arthritis', 'juvenile chronic arthritis', and more recently, 'idiopathic arthritis of childhood', The present article reviews the various classification criteria used for defining this group of disorders with emphasis on the common features as well as the major differences between these criteria. The major classes within this group with their characteristic clinical and laboratory features are also discussed. PMID- 10830004 TI - Clinico-immunological profile in juvenile rheumatoid arthritis--an Indian experience. AB - From a Pediatric Rheumatology Clinic 361 children diagnosed as juvenile rheumatoid arthritis (JRA) according to American Rheumatism Association-JRA criteria were studied retrospectively for their clinico-immunological profile. The mean age of onset in systemic, pauciarticular and polyarticular onset, JRA subtypes were 5.2, 6.8 and 7.2 years respectively. There was male preponderance in systemic and pauciarticular JRA. In seropositive polyarticular JRA, girls outnumbered boys. The frequency of occurrence of systemic, pauciarticular and polyarticular disease was 87 (24%), 108 (30%) and 166 (46%) respectively. The systemic onset disease was dominated by extra-articular manifestations in terms of fever (100%), rash (57%), hepatomegaly (51%) and lymphadenopathy (25%). The pauci- and polyarticular illnesses were commonly dominated by joint involvement, morning stiffness, and in few patients, by extra-articular manifestations also. The joints were involved symmetrically. Most commonly involved joints in order of decreasing frequency were knee, ankle, wrist and elbow in all the subtypes. Anemia and leucocytosis were observed in majority with higher frequency in systemic onset JRA. The rheumatoid factor (RF) was present in 15% of polyarticular JRA. RF was also present in 7 and 9% of patients with pauciarticular and systemic subtypes respectively. The antinuclear antibody was positive in only 3 out of 66 patients in whom the test was carried out. The demographic profile and trends in clinical features were similar to the studies reported on caucasian population with difference in the actual frequency of various clinical features. PMID- 10830003 TI - Juvenile rheumatoid arthritis--assessment. AB - The assessment of overall health status of a child with juvenile rheumatoid arthritis (JRA) is complex and multi-dimensional. The general physical examination is complemented by a rheumatological evaluation that includes determination of articular indices of inflammation and duration of inactivity stiffness. Laboratory assessment plays a critical role in monitoring side effects of pharmacologic management, but is limited in its ability to portray accurately the degree of active inflammation. Newly measureable indicators of inflammatory activity, such as serum cytokine and soluble cytokine receptors will likely become part of routine laboratory assessment in the future. Radiographs remain a useful tool for assessing disease progression, but may be replaced in the future by magnetic resonance imaging. In recent years, rheumatologists have realized that measurement of overall physical and psycho-social functional ability, quality of life, and pain are major descriptors to consider during routine follow up. They are also critical in the assessment of long-term clinical effectiveness. The importance of nutritional assessment has also been realized. This section seeks to describe some of the methodologic approaches currently used to assess the variables mentioned above, and includes a brief discussion of the evolving instrumentation which attempts to measure variables of a more cognitive or subjective aspect. PMID- 10830005 TI - Juvenile rheumatoid arthritis in India--rarity of antinuclear antibody and uveitis. AB - Juvenile chronic arthritis is a heterogeneous disease, having different subtypes. Among our 89 patients with juvenile chronic arthritis, we did not find even one patient with early onset pauciarticular disease with uveitis and antinuclear antibody positivity. Further, the prevalence of anti-nuclear antibodies and anti histone antibodies was very low whereas the prevalence of rheumatoid factor was similar to that reported from Western countries. Thus, the spectrum of juvenile chronic arthritis in India differs from that seen in the west. Larger population based studies of the disease are thus needed. PMID- 10830006 TI - Management of rheumatic diseases in children. AB - Rheumatic diseases are one of the common groups of chronic diseases of childhood. They are multifactorial in origin and tend to involve multiple organ systems. Consequently management of these diseases requires the expertise of many health and allied health professionals. This review article focuses on the medical management of three of the relatively common diseases: juvenile rheumatoid arthritis (JRA), systemic lupus erythematozus (SLE) and dermatomyositis (DM). PMID- 10830007 TI - Vasculitis in children. AB - Vasculitis in children is uncommon and hardly any information is available from India. We, at PGIMER, Chandigarh, have diagnosed and followed many children with vasculitis of different types though not all, which occur in children. In this article, we have given an overview of the vasculitides that we have encountered along-with a review of relevant literature. We have described 8 children with classical PAN and have highlighted a higher frequency of CNS involvement in our patients. Amongst the 10 BCPAN children, as many as 8 had peripheral gangrene which resulted in auto-amputation in 7. Gangrene of such severity has not been previously reported in this condition. We have also included 30 children with HSP. Gastrointestinal involvement was noted in 86.7% of children and in one of these, it was severe enough to result in hypovolemic shock. Such severe bleeding is very rare. Two of our patients with HSP came late to us after having been operated for an 'acute abdomen' elsewhere. Although renal involvement was seen less frequently than reported in the literature, the severity of involvement was greater (nephrotic range proteinuria in 62% and azotemia in 50%). We have only limited experience of Kawasaki Disease but it appears that children with this disorder are probably not being diagnosed in the acute stage in our country. PMID- 10830009 TI - Peripheral blood stem cell transplantation. PMID- 10830008 TI - Presentation and management of Helicobacter pylori infection in childhood. AB - Helicobacter pylori is responsible for one of the most frequently encountered infectious diseases worldwide. Helicobacter pylori infection can lead to the development of gastritis and peptic ulcer disease. The presence of Helicobacter pylori in the human stomach also represents an increased risk of gastric cancer and gastric lymphoma. Epidemiological data obtained in adults suggest that the actual colonization with Helicobacter pylori is in fact determined by childhood factors. Therefore, the pediatric age group represents the ideal target population for studies concerning the pathogenesis and epidemiology of Helicobacter pylori infection. The present work reflects our experience with regard to the diagnosis, epidemiology and pathogenesis of Helicobacter pylori infection in childhood. PMID- 10830010 TI - Fibroblast growth factor receptor mutations and craniosynostosis: three receptors, five syndromes. AB - The post eighteen months have been exciting time for craniosynostosis research. In a rapid flurry of publications, mutations of fibroblast growth factor receptors (FGFRs) have been identified in three of the best known craniosynostosis syndromes, namely Apert, Crouzon and Pfeiffer syndromes, as well as in Jackson-Weiss syndrome and thanatophoric dysplasia. These findings open many new avenues for craniosynostosis research including studies of diagnosis, pathogenesis, and mutagenesis. Here the major findings and their implications have been briefly reviewed. PMID- 10830011 TI - Perinatal mortality trends in a referral hospital. AB - A comparative study of perinatal mortality patterns over a period was conducted at a teaching hospital of South India. Among the 6,048 babies born from January 1984 to December 1985 (Group A), there were 265 (43.8/1000) still births and 127 (22.0/1000) early neonatal deaths. Three hundred and thirty seven (41/1000) babies were still born and 235 (29.8/1000) early neonatal deaths out of 8,215 deliveries during 1992-93 (Group B). The perinatal mortality rate (PMR) in Groups A and B were 57.9/1000 and 57.7/1000 respectively. Unbooked cases accounted for the majority (> 75%) of perinatal deaths during both the periods. The overall mortality rates in unbooked cases were three to four times higher than booked cases. Among the various causes of still births, antepartum haemorrhage and uterine rupture had increased. Septicaemia was the major cause of early neonatal deaths in Group A, but in Group B birth asphyxia and prematurity were the leading causes. Effective interventions like creating awareness among the target population to utilise maternal and child health services and early referral of high risk cases with improved intranatal and perinatal care can decrease the perinatal mortality. PMID- 10830012 TI - Under fives mortality in the urban slums of Lucknow. AB - The main objective of this study was to elicit proportional cause specific mortality in the underfives in the urban slums of Lucknow in North India. The families with under five mortality in the 28 randomly selected slums in 1993 were located from the records of the slum health workers and verbal autopsy was conducted to assign a cause of death. There were 71 deaths among 2796 children. The annual under five mortality was 25.4 and the under five mortality rate was 126.7. After the neonatal period, "high fever" that could not be classified into any other disease incorporated in the verbal autopsy instrument, was the most common symptom associated with death, seen in 21.1% cases (95% C.I.: 15.5-34.4%) followed by these diseases: pneumonia in 19.7%, diarrhea in 18.3% and measles in 11.4%. "High fever" as the leading symptom associated with death is being reported for the first time from the urban slums of India. There is an urgent need to identify the underlying etiologies of death due to "high fever" and the policy implications are that children with fever must receive immediate and continued medical attention till the symptom persists. PMID- 10830013 TI - HLA tissue types in nonresponders to hepatitis B vaccine. AB - Genetic factors are implicated in the response of normal subjects to hepatitis B vaccine. In order to investigate the immunogenetic factors associated with nonresponsiveness to hepatitis B vaccine, 93 health care workers were vaccinated with hepatitis B vaccine. Initial nonresponders (antibody not detected or antibody detected but < 10 mlU/ml) were revaccinated. Only 12 (12.9%) of the 93 health care workers, who had anti-HBs levels of 10 mlU/ml or less after revaccination were defined as absolute nonresponders. HLA typing were performed in these 12 nonresponders, Anti-HBs levels were determined by ELISA method in mlU/ml units. HLA-A,B,C,DR, and DQ typing was performed using the microcytotoxicity test. The HLA-A10 (pc less than 0.01) and CW4 (pc less than 0.006) were decreased whereas DR7 (pc less than 0.09) was increased in nonresponders. Although our initial results suggest the importance of genetic modulation of responsiveness to hepatitis B vaccine, a formal demonstration of the mode of inheritance of unresponsiveness to hepatitis B vaccine and the explanation of the role of genes in this matter will require further studies of families. PMID- 10830014 TI - Juvenile dermatomyositis. AB - The clinical profile of 7 children and their follow-up is described. There was female preponderance with mate to female ratio of 1:6. The median age of onset was 6 years. All the patients had skin rash, muscle weakness and abnormal enzyme profile. Muscle biopsy was performed in 6 and was abnormal in all of them. The electromyogram (EMG) was performed in 6 and was found abnormal in five. All the children responded well to corticosteroids. Two children received intravenous dexamethasone bolus and showed good response. PMID- 10830015 TI - Neonatal jaundice due to ABO incompatibility in Sri Lankan. AB - A prospective study was carried out on 101 neonates with jaundice due to ABO incompatibility. The direct Coomb's test was weakly positive in 4 cases. The indirect Coomb's test using the eluate was positive in 8 cases. In the maternal blood either IgG anti-A or anti-B haemolysin was present in high titre in every case. Phototherapy was given when the indirect serum bilirubin level exceeded 9 mg/dl. Exchange transfusion was done-in 39 cases, 9 babies requiring multiple exchanges. There were 2 deaths. PMID- 10830016 TI - Retrospective prediction of birth weight by growth velocity curves during neonatal period. AB - In this prospective study, birth weight of 304 babies born at Kamla Nehru Hospital Pune during study period was recorded. From these 304 babies, babies with birth weight above 2000 grams were selected (260 babies) to prepare growth velocity curves. Daily weight of these 260 babies was recorded for 30 days. The mean birth weight of study population was 2742.5 grams. Among the daily weight recorded babies, all the babies lost weight ranging from 92 to 218 grams (mean 121 grams) after birth. The weight loss continued upto 5 days. Days required to gain weight equal to birth weight ranged from 5 to 13 days. Total weight gain observed in 30 days was 734.7 grams. Predictive value of these curves was tested in 49 infants. Deviation upto 50 grams of predicted birth weight from actual birth weight was observed in 90% of babies on day-2, 79% on day-4, 65% on day-8 and 39% on day-30. PMID- 10830017 TI - Partial trisomy 18q. PMID- 10830018 TI - Primary pulmonary hypertension, Raynaud's phenomenon and generalised vasculitis in juvenile rheumatoid arthritis. PMID- 10830019 TI - Fanconi anemia. PMID- 10830020 TI - Endocrine aspects of Langerhans cell histiocytosis. PMID- 10830021 TI - Prescribing practices of pediatricians in malaria. PMID- 10830022 TI - Cardiac tamponade in juvenile rheumatoid arthritis. PMID- 10830023 TI - S. typhi infection in congenital nephrotic syndrome. PMID- 10830024 TI - Congenital hypertrophic pyloric stenosis at birth. PMID- 10830025 TI - Role of steroids in croup and beta agonists in bronchiolitis. AB - The administration of corticosteroids by various routes in the treatment of croup remains controversial as studies with varying designs and outcome measures have reported conflicting results. However, meta-analysis supports the use of steroids in croup. A dose equivalent to or greater than 100 mg/kg body weight of hydrocortisone, administered orally or parenterally appears to be effective. Nebulized budesonide in a dose of 1,000 micrograms, when dosages are given 30 minutes apart is also effective. The L-epinephrine is as effective as racemic epinephrine and should be used to relieve symptoms with or without steroids. Summarizing the value of beta agonists, for treatment of bronchiolitis is rather challenging. Beta agonists offer significant relief to only a subgroup of infants, with bronchiolitis, albeit, it remains a challenge to identify these infants prospectively. Availability of RSV immune globulin for prevention of RSV disease in high-risk infants and better defining the role of ribavirin for selected patients may offer additional avenues to the treating physician. PMID- 10830026 TI - Dexamethasone in bacterial meningitis: to use or not to use? AB - Permanent neurologic disabilities are seen in up to a quarter of survivors of bacterial meningitis despite major improvements in therapy. Experimental studies have demonstrated that most of the pathology in meningitis is mediated by inflammatory cytokines such as tumor necrosis factor (TNF) and interleukin-1 (IL 1), which are produced by host cells in response to bacterial invasion of the meninges. Dexamethasone has been used in a number of clinical trials to moderate the host response and to improve neurologic outcome of meningitis. Results of six randomized, placebo controlled trials are summarized in this review. Dexamethasone treatment did not lower mortality. Only a moderate, but not a significant reduction in the neurologic and audiologic sequelae was seen in dexamethasone recipients when Haemophilus influenzae type b (Hib) was the causative agent of meningitis. Following routine use of Hib vaccine, meningitis caused by this agent has virtually disappeared in the USA. Hence, findings from these trials may no longer be applicable in countries with high rates of immunization against Hib. Presently, there is little or no evidence showing a benefit of dexamethasone therapy in meningitis caused by S. pneumoniae or N. meningitidis. Global emergence of penicillin and cephalosporin resistant S. pneumoniae has raised new concerns about the use of dexamethasone in pneumococcal meningitis. Since dexamethasone significantly decreases the penetration and concentration of vancomycin and ceftriaxone in the CSF and delays CSF sterilization, adjunctive dexamethasone therapy may increase the risk of treatment failure in meningitis caused by antibiotic resistant pneumococci. An antibiotic combination should be used in the treatment of meningitis caused by antibiotic resistant pneumococci, particularly if dexamethasone is also being administered concurrently. PMID- 10830027 TI - Growth hormone therapy in children with short stature: is bigger better or achievable? AB - The introduction of recombinant DNA-synthesized human growth hormone in the mid 1980s, and its attendant unlimited supply, have led to wider application of growth hormone therapy in children. Over the past decade, the efficacy of growth hormone treatment in patients with Turner syndrome and chronic renal insufficiency, two conditions in which growth hormone secretion is normal, in improving growth velocity and final height, has also led to the consideration of growth hormone therapy in children with idiopathic short stature. Although thousands of patients with idiopathic short stature are currently being treated with growth hormone, the limited overall results available at this time do not show a significant improvement in final adult height despite an improvement in short-term growth velocity. Potential reasons for this outcome include 1) skeletal age advancing more rapidly than height age, 2) heterogeneity of the patient population comprising idiopathic short stature, 3) inherent inaccuracies of methodological tools, such as measurement of predicted adult height, and 4) a subset of children with idiopathic short stature who may, in fact, have partial growth hormone insensitivity. From a psychological perspective, the consensus of investigations in non-clinic-referred populations of psychosocial function in children with short stature do not indicate a disadvantage compared with children of normal height when socio-economic status is taken into consideration. These results, in conjunction with the minimal gains reported in behavioural measurements in idiopathic short children treated with growth hormone, question the traditional rationale that augmentation of growth velocity results in improvement in psychosocial well-being. PMID- 10830029 TI - Radiofrequency ablation in children. AB - Radiofrequency (RF) catheter ablation has ushered in a new era in the management of patients with symptomatic tachyarrhythmias. By providing the ability to cure the underlying arrhythmic substrate, RF catheter ablation obviates the need for life-long antiarrhythmic drugs. In the reported series, the success has been high and the complications have been infrequent and relatively minor. Not unexpectedly, RF catheter ablation has become the treatment of choice for patients with symptomatic paroxysmal tachyarrhythmias. The role of radiofrequency catheter ablation in infants and small children remains controversial, and awaits a larger experience and longer follow-up data. PMID- 10830028 TI - Antenatal steroids: miracle drug for preemies. AB - Antenatal corticosteroid administration has been unequivocally demonstrated to reduce neonatal morbidity and mortality. In addition to its well-documented role in reducing respiratory distress syndrome, evidence is accumulating indicating the global maturational effect of this therapy in the growing fetus. New data demonstrates, the hitherto relatively not well known, beneficial effects of this mode of therapy on renal, cardiovascular, gastrointestinal, endocrinal and cerebral maturation. Despite its well proven efficacy and safety, this therapy is used only in a small fraction of eligible mothers. In this article, biological basis of antenatal steroids use, clinical benefits including those to extremely premature infants, indications for their administration, treatment regimens, practice variations in their administration, cost analysis, and some directions for future research are reviewed. It is hoped that given the evidence reviewed in this article, the use of antenatal steroids administration in proper clinical settings will increase with a significant impact in reducing the neonatal morbidity and mortality. PMID- 10830031 TI - Therapeutic gastrointestinal endoscopy. AB - The safety, efficacy and utility of various therapeutic gastrointestinal (GI) endoscopic procedures performed on children (January 1992 to July 1995) at a tertiary referral centre in India were studied. A total of 1,030 sessions (upper GI 972 and lower GI 58) of therapeutic GI endoscopy were performed in 162 children (mean age 7.4 +/- 4 years; upper GI 115, lower GI 47). Various upper GI endoscopic procedures done were injection sclerotherapy (EIS), endoscopic variceal ligation (EVL), bougie dilatation of oesophageal strictures, balloon dilatations of oesophageal stricture/pyloric obstruction and retrieval of foreign bodies in 75%, 6%, 9%, 4% and 12% of children respectively. Therapy for bleeding from oesophageal varices constituted the major group (75%). Repeated EIS (sessions total--876, mean 8, range 5-15) performed on 86 children resulted in control of bleeding in all and eradication of oesophageal varices in 85% of cases. Minor complications (oesophageal ulcers and oesophageal strictures) due to EIS were observed in 9% of children. EVL (10 sessions in 7 children) was effective in controlling bleeding and substantial decrease of varices in all without any complication. Oesophageal dilation either by bougie (61 sessions in 10 children) or balloon (6 sessions in 3 children) were performed for benign strictures. Balloon dilatation of pyloric obstruction was successfully done in 2 children. Foreign bodies (retained or sharp) were retrieved from upper GI tract in 14 children. No complications were observed with stricture dilatation/foreign body retrieval. Therapeutic lower GI endoscopies were performed in 47 children (colonoscopic polypectomy in 92%, anal dilatation and piles banding in 4% each). One child with juvenile polyposis coli developed sigmoid colon performation following colonoscopic polypectomy which required surgical correction. We conclude that upper and lower GI endoscopic therapeutic procedures in children are highly effective and safe. The risk of major complication is very small in experienced hands and occasional minor complications, can be managed conservatively. PMID- 10830032 TI - Saudi aminoacidemias: a six-year study. AB - More than 2,500 samples were analyzed and we found 135 abnormal amino acid patients in different categories. The major four diseases are: Maple syrup urine disease (MSUD), Phenylketonuria (PKU) both classical PKU due to L-phenylalanine hydroxylase deficiency and 6-pyruvoyl tetrahydropterin synthase (6PTPS) deficiency, homocystinuria and Tyrosinemia. PMID- 10830030 TI - Nutrition and HIV infection in children. AB - HIV infection has profound effects on a patient's nutritional status because it can modulate appetite, nutrient absorption and basal metabolic rate. In addition, HIV infection can lead to the depletion of a variety of vitamins and micronutrients including vitamins A, D, B2, B6, B12, L-carnitine, iron, zinc and selenium. This review article summarizes existing data regarding nutritional defects in HIV-infected patients and the results of clinical studies addressing the effects of nutritional supplementation in infected patients. PMID- 10830033 TI - Infantile pseudotumor cerebri related to viral illness. AB - Pseudotumor cerebri (PTC) presents with varied spectrum of manifestations, association with diverse pathological conditions and variable outcome. A preliminary observation on children presenting with PTC is being presented because of certain peculiarities, not observed earlier, like occurring in clusters during particular season, all were infants having close relationship with viral illness and all had extremely benign outcome. Children presenting with raised ICP were hospitalised and evaluated. The relevant investigations carried out included lumbar puncture, cranial ultrasonography, CT Scan and culture studies. Fifteen children, all in infancy with male-female ratio of 2:3 presented in clusters of 11 (73.3%) cases during late summer and early fall presented with bulging fontanelle, vomiting 11, fever 12, respiratory infection 5, diarrhea 5, lateral deviation of eye balls 4 and skin rashes 5 cases. Contact with viral illness in the family was found in 10 cases. Lumbar puncture showed high opening high CSF pressure in all. Results of this study suggest peculiar and perhaps different form of PTC seen recently which affected only the infants. These were closely related to viral illness and had benign outcome. PMID- 10830034 TI - Spectrum of congenital anomalies associated with biliary atresia. AB - Several congenital anomalies have been reported in association with biliary atresia. We have analysed the type and frequency of anomalies observed over a 10 year period in consecutive patients operated for extrahepatic biliary atresia at our institution. Of the 107 infants who underwent a laparotomy and surgical correction of biliary atresia, 9 (8.4%) showed significant associated anomalies. Among them, 5 (55.5%) had splenic malformations, 6 (66.6%) had digestive anomalies in the form of malrotation, Meckel's diverticulum and jejunal atresia. One patient had the classical polysplenia syndrome. Our follow-up period was limited (i.e. 5 months) during which time 3 of the 9 (33.3%) patients with associated anomalies became jaundice-free. A higher incidence of malformations found in association with biliary atresia supports the congenital theory and a more thorough search for these anomalies is recommended. PMID- 10830035 TI - Zinc tolerance test patterns in normal children and in moderate and severe zinc deficiency states. AB - Twenty-six normal, 38 moderately and 14 severely zinc-deficient children, aged 2 12 years, were examined by clinical and laboratory approaches. After fasting blood sampling, 120 mg zinc sulphate (25 mg elemental zinc) were administered orally to each group of children, to obtain zinc tolerance curve patterns. Sampling proceeded to the 2nd and 4th hours of the loading-test period. Plasma zinc was assessed on an atomic absorption spectrophotometer. In normal children, at the 2nd hour of loading, a significant (p < .001) elevation (1.764 +/- 0.133 mg/l) in the mean (+/- SEM) plasma zinc level was noted; also at the 4th hour a significant (p < .001) decrease (1.506 +/- 0.123 mg/l) in the mean plasma zinc level was shown. The mean plasma zinc level at the 4th hour was found higher than the mean fasting plasma zinc level (1.054 +/- 0.061 mg/l), but lower than the mean level found at the 2nd hour. In moderately zinc-deficient children, the rise in the 2nd hour and the fall in the 4th hour in the plasma zinc level were highly significant (p < .001 and p < .001, respectively) in relation to fasting blood level. However, in severely zinc-deficient children, the intensity of the increase (0.746 +/- 0.147 mg/l) in plasma zinc level at the 2nd hour was of lesser significance (p < .006) and the fall (0.424 +/- 0.061) mg/l) at the 4th hour was not significant. Therefore, in children with normal plasma zinc levels, an increase of more than 0.50 mg/l was seen at the 2nd hour of loading. This rise was seen to persist at the 4th hour. However, in children with moderate zinc deficiency, although again an increase of 0.50 mg/l was seen at the 2nd hour this increase did not persist at the 4th hour; and the 4th-hour value showed a significant decrease in relation to the 2nd hour value. Whereas, in children with severe zinc deficiency the rise of plasma level at the 2nd hour was less than 0.5 mg/l and the fall at the 4th hour was to such a level which was not significant in relation to fasting zinc level. This could be due to enhanced uptake of zinc off the circulation by the depleted tissues in severe zinc deficiency. PMID- 10830036 TI - Does B.C.G. vaccination prevent or postpone the occurrence of tuberculous meningitis? AB - The reported efficiency of B.C.G. vaccine in prevention of primary childhood or sputum positive adult tuberculosis vary widely but it is believed to offer significant protection against hematogenous forms of tuberculosis. The present hospital-based, case control study was aimed at evaluation of impact of B.C.G. vaccination on one of the hematogenous forms of tuberculosis i.e. Tuberculous meningitis (TBM). Records of cases of TBM admitted in the pediatric wards of Lok Nayak Hospital, Delhi, over one year were analyzed retrospectively for B.C.G. positivity status and age distribution. The results were compared with similar data obtained from a control group comprising of non-TBM cases in pediatric wards of same hospital, recruited on 2 days, separated by an interval of three weeks. No statistically significant difference was found in B.C.G. positivity status of cases and control groups. However, there was an obvious trend in age distribution among cases of TBM with respect to the B.C.G. status, with a significantly higher proportion of B.C.G. vaccinated children in over 5-year age group, as compared to those in under 5 year age group. The results, therefore, indicate probable effectiveness of B.C.G. against TBM only for a limited duration i.e. single B.C.G. immunization only postpones rather than prevent the occurrence of TBM. PMID- 10830038 TI - Interstitial lung disease. AB - Interstitial lung disease in children is a complex group of disorders whose etiology and pathogenesis is not entirely clear. Although the basic pathogenesis has been extrapolated from adult studies, its relevance in the pediatric population can be questioned. Several classifications of this condition have been put forward but Liebow's histologic classification is well accepted. Clinical manifestations are non-specific and can vary from no symptoms and a positive chest X-ray to more characteristic signs and symptoms. The triad of tachypnea, intercostal retractions and dry crackles is characteristic. Chest roentgenograms are a useful diagnostic tool and CT scans help to stage disease severity. Spirometry classically shows a restrictive pattern. Lung biopsy is the gold standard for establishing a diagnosis. Corticosteroids are the cornerstone of treatment in this condition. Thus, in view of the multifactorial nature of this disease, more extensive multi-centre prospective studies are required in order to better understand this rather complex group of disorders. PMID- 10830037 TI - Upper airway obstruction. AB - The airways in infants and children are anatomically different from adults, thus predisposing them to more acute upper airway obstruction. The causes of upper airway obstruction may be infective or non-infective. The presence of dysphonia, dysphagia, abnormal respiratory pattern, cough and abnormal posture suggests upper airway obstruction. The general management consist of supportive care with minimal invasive procedures. The specific treatment depends on the causes and is discussed in text. PMID- 10830039 TI - Defective growth hormone secretion and hypogonadism in the new syndrome of congenital hypoparathyroidism, growth failure and dysmorphic features. AB - A child with extreme growth failure, dysmorphic features, hypoparathyroidism, and abnormal skeletal survey was studied. He was a product of first degree consaguineous marriage who had intrauterine growth retardation and presented at 14 days of age with hypocalcemic tetany with normal cardiovascular system and immune function. Endocrine evaluation after infancy revealed defective growth hormone (GH) secretion in 2 provocation tests and lack of clinical and testosterone response to human chorionic gonadotrophin (HCG) therapy. PMID- 10830040 TI - Antemortem diagnosis of Leigh's disease: role of magnetic resonance studies. AB - A two-and-a-half-year-old male child presented with recurrent attacks of intractable vomiting, psychomotor retardation since 14 months of age. He had also lower cranial nerve palsy and corticospinal involvement. Magnetic resonance imaging had shown multiple well circumscribed areas of hypointusity in T1 weighted image which were brightly hyperintense in heavily T2-weighted image. The lesions were seen in basal ganglia, thalamii and brainstem and spared mamillary bodies. Magnetic resonance spectroscopy demonstrated lactate peak in the affected areas confirming the diagnosis of Leigh's disease. The child responded well to large dose of vitamin 'B' therapy. PMID- 10830041 TI - Gastric perforation secondary to recurrent trichobezoar. AB - A rare case of 10-year-old female child with recurrent trichobezoar stomach is reported, which presented with features of gastric outlet obstruction with perforation. PMID- 10830042 TI - A very rare type of multiple intestinal atresias: "a string of pearls". AB - Intestinal atresia is the most frequently encountered cause of ileus in the neonate. Of all atresias combination of small and large bowel atresias is extremely rare. In 1973, Guttman presented a case with multiple, widespread atresias of small and large bowel, intraluminal calcifications and a hereditary nature. This paper describes the detailed pathological findings of a similar case of multiple intestinal atresias and discusses for the pathogenesis of this rare condition. PMID- 10830043 TI - Primary aortoesophageal fistula: presenting as massive upper gastrointestinal hemorrhage. AB - Primary aortoesophageal fistula is a rare cause of upper gastrointestinal bleeding. A six-year-old boy presented with massive upper gastrointestinal hemorrhage. Endoscopy revealed a submucosal bulge in the esophagus with an ulcer and clot at the top. Lateral skiagram of the chest showed a posterior mediastinal mass. CT scan of the chest revealed a ruptured aortic aneurysm into the oesophagus, confirmed the diagnosis. The patient succumbed to the illness before he could be subjected to definitive treatment. PMID- 10830044 TI - Capillaria hepatica parasitism. AB - Capillaria hepatica is rarely encountered in humans, with fewer than 30 documented cases. The clinico-pathological features of capillaria hepatica infection, diagnosed on liver biopsy of a 6-year-old child are discussed. Pathologically, it is characterised by prominent granulomatous lesions in the liver surrounding the eggs, which on cursory examination may be confused with Schistosoma mansoni. PMID- 10830045 TI - Transient right sided hypertrophic cardiomyopathy in an infant born to a diabetic mother. AB - Hypertrophic cardiomyopathy (HCM) is a rare primary myocardial disease, characterized by hypertrophy of the left and/or right ventricle. Infants of diabetic mothers (IDM) are at risk for development of HCM, respiratory distress and persistent pulmonary hypertension. A case of severe right sided HCM in an infant born to a diabetic mother is presented. The patient's findings were complementary to the previous observations reporting HCM in IDM. The presence of disproportionate septal hypertrophy in the echocardiography of an infant born to a diabetic mother is highly suggestive of HCM in IDM. In our opinion, further cardiac evaluation is not indicated unless other cardiac abnormalities are suspected. PMID- 10830046 TI - Neonatal empyema thoracis. AB - Empyema thoracis, a serious complication of pneumonia, fortunately remains a less common cause of respiratory distress in neonates. Only 14 cases of neonatal empyema thoracis have been described in the world literature. The condition is characterized by its rarity, inability to identify any consistent predisposing factors, uncertain pathogenesis, rapid course, lack of consensus on management and a high mortality. We describe here two cases of empyema aged 6 and 8 days caused by E. Coli and Klebsiella respectively. Out of them one survived. A brief review of literature follows the above account. PMID- 10830047 TI - Huge omental cyst mimicking ascites. AB - An unusual case of omental cyst is described. A 4-year-old child presenting as abdominal distension of short duration, clinically diagnosed as ascites, was subsequently proved to be giant omental cyst. The case is reported because of its unique presentation. PMID- 10830048 TI - Management of Wiskott-Aldrich syndrome. AB - A five-month-old boy presented with lower gastrointestinal bleed, recurrent infections and eczema. Blood picture revealed small platelets, high IgA, and IgM levels. A diagnosis of Wiskott-Aldrich Syndrome was made. The recent concepts in molecular pathology of the disease and treatment are discussed. PMID- 10830049 TI - Improving the care of sick young infants: a challenge for the near future. PMID- 10830050 TI - An update of pediatric surgery. PMID- 10830051 TI - Biliary atresia--the current management. AB - Despite extensive research, controversies still exist regarding the etiology, pathology and management of biliary atresia. It is now thought to be a progressive panductal inflammatory obliterative process and not a developmental anomaly. The histologic changes are indistinguishable from neonatal hepatitis but some changes have prognostic significance. The clinical presentation is that of infantile obstructive cholangiopathy-waxing and waning icterus, clay coloured stools and high coloured urine from early neonatal period. The diagnosis is suggested by the absence of intestinal excretion on HIDA scan and confirmed on operative cholangiogram. Of utmost importance towards the final prognosis is early detection, prompt confirmation and surgical treatment before 2 months of age. Even with early treatment the result of bilioenteric drainage procedures have been discouraging in the long term. Portoenterostomy (PE) done in older children has been largely unsuccessful all over the world. The poor results of PE prompted the search for alternative treatment and liver transplantation (LT) has emerged as a viable treatment option both as a primary procedure and after failed PE. Although the technical know-how and infrastructure are available, LT in children has still not been done in India because of various economic and social constraints. It is hoped that all physicians and surgeons dealing with such patients would also consider this treatment modality. PMID- 10830052 TI - Prevalence of late orchidopexy is consistent with some undescended testes being acquired. AB - To diagnose the incidence of orchidopexy versus age over a 15-year period, a study was conducted of all patients discharged from a single institution for orchidopexy with reference to age during operation. The hypothesis drawn was that some boys have acquired UDT and therefore, will present late despite recommendations for early diagnosis and treatment. The study was conducted on patients from Royal Children's Hospital, Melbourne (1980-94). The results suggested that while the optimal age for management of congenital UDT has been lowered to one to two years of age by under-graduate education, the persistence of a significant number of older children undergoing surgery suggests that some UDT's are acquired. It also showed that the proportion of orchidopexies performed in infancy increased over the 15-year period while the proportion performed in late childhood remained constant. PMID- 10830053 TI - Role of extramucosal seromuscular biopsy during preliminary fecal diversion in the management of Hirschsprung's disease. AB - The routine use of frozen section biopsy to identify distal ganglionic bowel in Hirschsprung's disease is the accepted norm in well developed centers. With the aim of overcoming non-availability of frozen section biopsy and reducing the duration of definitive surgery, an alternative algorithm, based on extramucosal seromuscular biopsy at the time of preliminary fecal diversion and its examination by routine paraffin sections and staining, was developed and used in 100 patients at our centre. This technique accurately marked the ganglionic colon for pull-through operation in 96.2% of cases with rectosigmoid transition zone and in 91.7% of those with transition zone in the descending colon, thus obviating the need for frozen section biopsy and saving nearly one hour during the pull-through surgery. However, in the more proximal varieties of Hirschsprung's disease frozen section biopsy was still considered to be indispensable. PMID- 10830054 TI - A practical approach to the investigation and management of long gap oesophageal atresia. AB - Oesophageal atresia in which there is a long gap between the ends of the oesophagus remains one of the most difficult problems confronting the pediatric surgeon. While it is generally accepted that the best results are obtained if oesophageal continuity is established, this may be difficult to achieve. Various manoeuvres have been advocated to mobilise and lengthen the oesophagus, and if these measures fail, cervical oesophagostomy and oesophageal replacement may become necessary. This paper outlines the situations in which long gap oesophageal atresia might be expected, how infants should be investigated to determine the most appropriate type and time of procedure, and provides a basis for surgical decision-making during the operative procedure itself. These guidelines should enable oesophageal continuity to be obtained in the vast majority of infants with long gap oesophageal atresia. PMID- 10830055 TI - Influence of associated anomalies on the management of oesophageal atresia. AB - Associated congenital abnormalities occur in over 50% of infants with oesophageal atresia. Many of them influence the way in which the oesophageal atresia is managed and therefore should be detected as soon as possible after birth. Early recognition of those with no prospect of long-term survival (e.g. Trisomy 18, bilateral renal agenesis) will avoid unnecessary surgery to the oesophagus. About 12% of oesophageal atresia infants will have no surgery performed. This paper presents a protocol for the initial investigation and management of associated anomalies in oesophageal atresia, based on an experience of 670 infants treated at the Royal Children's Hospital, Melbourne, Pre-operative detection of associated anomalies of oesophageal atresia requires clinical examination, pre operative renal ultrasound (if the child has not passed urine), and an echocardiograph of the heart to exclude duct-dependent lesions. If there is clinical suspicion of a major chromosomal abnormality, surgery should be deferred until chromosomal analysis has been completed. PMID- 10830056 TI - Pediatric liver transplantation: the Indian perspective. AB - Liver transplantation is now the accepted treatment for end stage liver disease, hepatic-based inborn errors of metabolism and localised primary hepatic malignancies. In children the commonest indication is biliary atresia. Innovations in surgical techniques, particularly liver reductions/splits and living related transplants have not only shortened the waiting period but have improved the results of surgery as well. The long term survival for pediatric liver transplant recipients in most large series from the Western world is more than 85%. Treatment protocols for ITU care of donors and recipients, anaesthetic care during surgery and post-operative immunosuppression are available. A country like India has a crying need for the development of facilities for liver transplantation in children but a number of factors have hampered the progress. The cost of surgery and post-operative immunosuppression and attitudes on life and death issues have been the greatest impediment. A few organisational details have also to be addressed to in order to minimise interdepartmental clashes of interest. It is hoped that a viable programme would soon be available for the children in India. PMID- 10830057 TI - Current status of small bowel transplantation in children. AB - Small bowel transplantation is gradually changing from an experimental procedure to a very desirable and viable treatment option in children with irreversible intestinal failure due to either short bowel syndrome or functional impairment. Long term total parenteral nutrition and home parenteral nutrition would be necessary to manage these children in the absence of a small bowel transplant programme. Parenteral nutrition is also associated with complications which can result in chronic liver disease. In India, there is no infrastructure for this treatment option and even if it was there the cost of this method of treatment is likely to be more than the cost of post-operative immunosuppression. Small bowel can be transplanted as an isolated graft, in combination with the liver or as part of a multivisceral transplant. The operative techniques have been standardised. Major post-operative complications result from sepsis and lymphoproliferative diseases. The best results have been obtained with a combined liver and small bowel transplant. PMID- 10830058 TI - Timing of surgery for common pediatric surgical conditions. AB - Depending upon various factors the surgical procedures in childhood are divided into three groups--immediate, intermediate and elective. The timing of surgery is probably the most important factor governing the outcome of surgical correction in pediatric surgery. With continuing research and clinical experience our understanding of the various conditions have improved and with that has come a change in the optimum timing of many surgical procedures. This paper highlights the best timing of surgery for some of the commoner pediatric surgical procedures and the reasons behind these so that the children may be referred to pediatric surgeons in time. PMID- 10830059 TI - Fluconazole. AB - Incidence of systemic candidiasis is increasing with improvement in the survival of high risk neonates who undergo multiple interventions. Therapies available to treat systemic fungal infection are few and have several drawbacks. Fluconazole, a new triazole derivative may be a useful anti-fungal agent in view of its excellent oral absorption, easy administration, low plasma protein affinity, long half-life, high concentrations in urine and CSF, minimal adverse reactions, wide spectrum of anti-fungal activity and high specificity for fungal cytochrome P450 system. Its utility in neonates and children with candidiasis has already been documented by few case reports and studies. PMID- 10830060 TI - Hepatitis A in day care centre. AB - A focal outbreak of hepatitis was detected in a day-care centre for children centrally located in Pune. The source of infection was suspected to be an 11-year old child who probably got the infection from his school. Seven out of 15 children from day-care centre developed clinical hepatitis. Two cases of secondary infection were identified among the family contacts of infected children. Sera from all the nine sick children were positive for anti-hepatitis A virus-IgM antibodies. A stool sample from a case of secondary infection showed presence of HAV-RNA by RT-nested PCR. These findings proved that the outbreak was caused by hepatitis A virus. PMID- 10830061 TI - Early neonatal mortality in an intramural birth cohort at a tertiary care hospital. AB - Early neonatal mortality (ENM) occurring among 12,283 consecutive live births over a period of 3 years were analysed. The early neonatal mortality rate (ENMR) was 26.6/1000 live births. Birth weight less than 2,000 gm, lack of antenatal care, male sex, operative vaginal delivery, prematurity and multiple pregnancy were significantly associated with early neonatal deaths. Birth asphyxia was found to be the most important cause of death, followed by hyaline membrane disease and congenital malformations. Majority of the asphyxia related deaths were due to late intrapartum referral of the mothers. Forty-two per cent of early neonatal deaths occurred in babies weighing less than 1,500 gm. Early identification and referral of high risk mothers and health education would significantly reduce the early neonatal deaths. PMID- 10830062 TI - Age during breast feeding and timely suckling. AB - A cross sectional study of 398 mothers and infants was conducted to assess the age at first breast feeding, and its determinants, in Shimla. We found that the timely first suckling rate was 0.10 in urban group and 0.11 in rural group. The median age at initiation of breast feeding was five hours among urban cases and four hours among rural cases. Most of the infants were breast fed by the age of 24 hours which is a positive sign and is higher than previous studies. Absence of definite trend vs mother's education may be due to other factors. A community based educational programme to promote early breast feeding is required, along with training on lactation management counselling for health workers. This will help in achieving healthy child development through prevention of childhood malnutrition. PMID- 10830063 TI - Influence of parental literacy and socio-economic status on infant mortality. AB - The influence of parental literacy and socio-economic status (SES) on infant mortality rate (IMR) was studied. This report is only a part of much larger community-based prospective study conducted to determine the factors influencing infant mortality in Dakshina Kannada district of Karnataka, by the Department of Community Medicine, Kasturba Medical College, Manipal, with the financial assistance form IDRC, Canada. A total of 12,857 livebirths during the period of October 1, 1991, to September 30, 1992, where registered and followed up for one year. There were 391 infant deaths which gives an IMR of 30.4 per 1,000 livebirths. Univariate analysis and subsequent stepwise multiple logistic regression analysis revealed that, literacy and SES have a very significant role in reducing IMR. PMID- 10830064 TI - Congenital lobar emphysema: a review of 10 cases. AB - We reviewed the clinical features and management of 10 patients with congenital lobar emphysema (CLE) treated in our institution over a period of seven years, from September 1988 to September 1995. The age of onset of symptoms ranged from few days from birth to six months. Correct diagnosis was made in only 4 patients before referral, even though diagnosis could have been arrived at from the chest radiograph in all of them. Lobectomy was done in seven patients, while one patient had vascular ring resected. Of the three patients managed conservatively, in one lobectomy was performed at the end of 5 months conservative course as her clinical condition deteriorated during an intercurrent chest infection. Of the remaining two, one has mild episodic reactive air way disease and the other patient is asymptomatic. PMID- 10830065 TI - Systemic air embolism in respiratory distress syndrome. AB - We report a case of severe respiratory distress syndrome which required intermittent positive pressure ventilation and led to severe pulmonary interstitial emphysema (PIE) and massive air embolism. PMID- 10830066 TI - Fetal Ebstein anomaly--a rare cause of non-immune hydrops. AB - A rare case of fetal Ebstein anomaly presenting as non-immune hydrops fetalis at 28 weeks of gestation is reported along with its review of literature. PMID- 10830067 TI - Mummified insect as foreign body in the respiratory tract. AB - A case report of a child with a cockroach in bronchus is described. PMID- 10830068 TI - Craniosynostosis and Klippel-Feil syndrome: a rare association. PMID- 10830069 TI - Dyskeratosis congenita (Zinsser-Engman-Cole syndrome). PMID- 10830070 TI - Piercing foreign body in neonate: a case report. PMID- 10830071 TI - [Euthanasia in Europe--give us alternatives]. PMID- 10830072 TI - [The complement system: an old story or target of new therapeutic approaches?]. AB - The complement system is a multifactorial protein cascade system which is essentially involved in the early unspecific immune response. Its major function is the activation of cellular defense mechanisms, opsonisation of foreign particles and the destruction of target cells. While the impact of the different complement components for bacterial elimination still remains controversial, overwhelming activation of the complement cascade, however, can induce life threatening tissue damage due to the effective cytotoxic properties. In the last years a variety of studies demonstrated beneficial, organ protective effects of complement modulation in models of severe inflammation. Attempts to control the complement system include the application of endogenous complement inhibitors e.g. C1-inhibitor (C1-INH) or the administration of recombinant complement receptors such as the soluble complement receptor 1 (rsCR1). Moreover antibodies against key proteins (C3, C5), against their activation products (C5a) or against complement receptor 3 (CR3, CD18/11b) mediated adhesion of leukocytes to the vascular endothelium, represent effective options of complement modulation. Besides this, insertion of membrane bound human complement regulators (DAF- CD55, MCP- CD46 or CD59) into xenogenic donor organs has proven effectiveness to prevent xenograft rejection. The described interventions protected from severe organ damage in various animal models of sepsis, myocardial and intestinal ischaemia-reperfusion injury, ARDS, nephritis, and xenograft rejection. With respect to recent clinical data, complement inhibition could represent a useful therapeutic strategy to control overwhelming inflammation. Own experiments demonstrated protective effects of complement modulation with C1 INH and rsCR1 in a model of complement induced pulmonary injury. With respect to sufficient host defense, however, the use of complement inhibitors must be considered carefully. PMID- 10830073 TI - [Relation between oxygen consumption and cardiac output during inhalation anesthesia under the influence of catecholamines. A study in dogs]. AB - OBJECTIVE: The metabolic regulation of tissue blood flow manifests itself in a linear relation between blood flow and oxygen consumption (VO2). It is unknown, however, if this fundamental physiological principle operates also during inhalation anaesthesia and catecholamine therapy, both known to be associated with changes of cardiac output (Q) and VO2 in opposite directions. METHODS: On different days, 17 trained, healthy dogs (26-33 kg) with chronically implanted flow probes around the pulmonary artery were either anaesthetized with halothane, enflurane, isoflurane, sevoflurane, and desflurane at increasing minimum alveolar concentrations (1-3 MAC) or treated with one of the endogenous catecholamines epinephrine and norepinephrine or the synthetic ones dobutamine and dopexamine (a total of 186 experiments). Cardiac Output (ultrasound transit-time flowmeter) and VO2 (indirect calorimetry) were measured continuously and the relations between both variables (Q/VO2 relations) analyzed. MAIN RESULTS: In awake dogs under basal metabolic conditions, VO2 was 4.6 +/- 0.1 ml x kg-1 x min-1 and Q 105 +/- 3 ml x kg-1 x min-1 (mean +/- SEM). During inhalation anaesthesia, VO2 and Q decreased in parallel, yielding a uniform Q/VO2 relation, which was nearly linear. Above 2 MAC, O2 extraction increased by 50%, indicating compromised oxygen delivery (DO2). Imposed increases in Q and thus DO2 during anaesthesia to rates comparable to that in the awake state did not restore VO2 to baseline. Catecholamines increased both VO2 and Q in a dose-dependent manner, albeit to a different extent. The resulting Q/VO2 relations were linear up to the maximum effects, but their slopes increased about threefold in the order norepinephrine (34), epinephrine (54), dobutamine (86), and dopexamine (105). Despite these differences, VO2 and Q correlated linearly over the whole range studied, which covered a doubling of VO2 and an up to fourfold increase in Q. CONCLUSIONS: The metabolic regulation of blood flow apparently also operates during inhalation anaesthesia, regardless of the anaesthetic. Catecholamines (except norepinephrine) preferentially stimulated Q rather than VO2 (excess perfusion), so that agents like dopexamine might be preferred in the treatment of low cardiac output states because of its low metabolic costs. Our observations imply that cardiac output alone provides little information on the function of the circulation during inhalation anaesthesia and catecholamine therapy unless related to metabolic demands, i.e. to VO2. PMID- 10830074 TI - [Euthanasia in Europe--ten countries with special consideration of the Netherlands and Germany]. AB - This article presents an overview of the current situation of euthanasia in Europe. Emphasis is given to the positions discussed in the Netherlands and in Germany. The current situation, the development of the legal positions, and the resulting debate are established by analysing English and German anesthesiological and medical-ethical journals. It has to be noted that many physicians are not satisfied with the terminology of euthanasia. The traditional concepts of euthanasia do not cover the aspect of accompanying terminally ill persons until they have died. The differentiation of active, passive, and indirect euthanasia does not correspond to the practical handling of the problem. Many physicians are in need of an open discussion of euthanasia-related issues. The way euthanasia is practiced in the Netherlands has strongly influenced the further development of the debate in Europe. Even though the Dutch model is rejected by the jurisdications of virtually all other countries, and official statements of medical corporations stick to the disapproval of active euthanasia, studies examining the attitudes towards euthanasia and the treatment of it in daily routine show that active interventions to shorten life are performed to different degrees outside of the Netherlands as well. PMID- 10830075 TI - [Desflurane--general anesthesia for cesarean section compared with isoflurane and epidural anesthesia]. AB - Desflurane 2.5% was compared to Isoflurane 0.5% in a randomized study in terms of maternal and newborn effect on both groups with epidural anesthesia. Fifty patients under general anesthesia were randomly designated to receive either desflurane 2.5% or isoflurane 0.5% maintained in a 50-50% nitrous oxide and oxygen mixture. Twenty-five patients were assigned to receive epidural anesthesia using 15 ml ropivacaine 7.5 mg/ml with fentanyl 100 micrograms. Intraoperative hemodynamic changes, blood loss and maternal awareness were recorded. Apgar scores at 1 and 5 min., neurologic adaptive capacity scores (NACS) at 2 and 24 hours and umbilical vein blood gas analysis were done to assess the neonatal outcome. Intraoperatively, heart rate and blood pressure changes were similar in both desflurane and Isoflurane group at 0.4% MAC (minimal alveolar concentration). Blood loss and arterial blood gas analysis were not problematic and did not differ significantly between the three groups. In the desflurane group, the patients were more easily awake and cooperative compared to the isoflurane group. The patients were interviewed about intraoperative awareness 24 and 48 h after the operation. None of them reported awareness during the operation. Similarly, the level of postoperative comfort was the same in both groups. Comparing the general and epidural anesthetic groups, no differences could be detected in neonatal outcomes. Conclusion is that there is one significant difference between desflurane 2.5% and isoflurane 0.5% anesthesia for cesarean section and it is the rapid recovery characteristic with desflurane which makes it an attractive alternative to TIVA (total intravenous anesthesia) and to other inhalational anesthetics available to obstetric anesthesiologists. PMID- 10830076 TI - [Self-reported substance abuse related emergencies: frequency and nature]. AB - OBJECTIVE: The aim of this study was to estimate the frequency and nature of self reported and drug-related emergencies. METHODS: 47 patients of a ward for opiate detoxification were interviewed about their experiences with drug-related emergencies. Typical categories had to be found like overdoses, seizures, accidents and suicide attempts respectively. RESULTS: 68% had own experience with drug-related emergency. A majority suffered opiate overdose with different extensions as unconsciousness or breath-depression. Alcohol and polydrug use was associated with overdose. Drug-related accidents were only reported by men. Half the number of drug-related emergencies were treated in hospital. Most emergencies occurred alone either in a home environment or outside. CONCLUSION: Harm reduction interventions like observed user rooms should be established. Furthermore other strategies to reduce the number of emergencies as sharing naloxon or resuscitation programs in wards for detoxification could also be an effective method to prevent near fatal or fatal overdoses in dependent subjects. PMID- 10830077 TI - [Standardized and complementary measures in pain therapy]. PMID- 10830078 TI - [The etiologic role of repressed expressivity in headache]. PMID- 10830079 TI - [Music and anesthesia in pain therapy]. PMID- 10830080 TI - [Acute traumatic myocardial infarction with cardiogenic shock in severe polytrauma--a case report]. AB - A 41-year-old man suffered severe polytrauma and developed a traumatic myocardial infarction with cardiogenic shock. Thrombolysis as well as coronary bypass grafting was contraindicated due to accompanying injuries. An attempted early coronary revascularization by percutaneous transluminal coronary angioplasty (PTCA) failed due to dissection of the left interventricular coronary artery. Treatment of cardiac insufficiency was complicated by intraabdominal haemorrhage enforcing emergency laparotomy. Intraaortic balloon counterpulsation proved to be efficient in supporting circulation in these circumstances. The case report documents the practicability and importance of treating both myocardial ischaemia and attending injuries in an equivalent and coordinated manner in traumatic myocardial infarction. PMID- 10830081 TI - [Facts and controversies in the treatment of cardiac insufficiency]. PMID- 10830082 TI - [Results of large-scale clinical trials and medical practice]. AB - The results of large scale clinical trials are part of the scientific data on which medical and, in particular therapeutic decisions, are base; very concordant data may even be used to define appropriate or inappropriate prescribing behaviour in the context of recommendations or references of good medical practice. Large scale clinical trials, however, have limitations inherent to their method; they give a mean result observed in an average population. The true question is to know if the therapeutic benefit proved in a therapeutic trial will be observed in a given individual in the future. If the aim of treatment is to make a symptom disappear or act on an objective intermediate criterion related to the prognosis, the benefits for the individual are easy to assess. When the aim is to prolong life or prevent a recurrence of a serious illness, the probability of avoiding a complication by treatment is only a statistic. This probability depends o: the amplitude of the therapeutic benefit observed in the trials; the resemblance of the patient to the "mean population" included in trials; the comparison of the treatment of the patient with the "mean treatment" used in the trials. An exact understanding of scientifically proven data then encounters the limits of the capacity of human memory. Probability medicine remains, however, the method which enables, for a given patient, the choice of treatment with the greatest chance of being effective, providing the elementary rules of utilisation of the information are respected. PMID- 10830083 TI - [Cardiac insufficiency: what treatment? what dose? For which patients? Converting enzyme inhibitors and diuretics]. AB - Diuretics were the first family of drugs to be used in cardiac failure. They improve symptoms but no randomised control trials show their efficacy in prolonging survival. The results of the recent RALES trial, however, provides evidence in favour of antialdosterone diuretics on survival in association with a loop diuretic and an angiotensin converting enzyme inhibitor. At present, the legal requirements in France state that "the association of spironolactone and converting enzyme inhibitors is possible with low doses of angiotensin converting enzyme inhibitors and diuretic hypokalaemiant; kalaemia and creatinine have to be monitored". "The association of hypokalaemia-inducing diuretics (loop diuretics, thiazides and similar: the association with this type of diuretic, rational and useful in certain patients, does not exclude the risk of hypo- or even hyperkalaemia, especially in renal failure and diabetes; it also imposes the monitoring of serum potassium and eventually of the electrocardiogram and, if necessary, to reconsider the treatment". Many points remain unclear concerning the value and harmlessness of the prescription of diuretics in asymptomatic left ventricular failure. In cases of diuretic resistance, the use of intravenous administration, the fragmentation of doses or the association of diuretics, may induce a diuretic response. Angiotensin converting enzyme inhibitors are the first line treatment of moderate and severe cardiac failure and in post infarction left ventricular dysfunction. On the other hand, the value of this family of drugs in left ventricular failure and normal systolic function has not been demonstrated. Analysis of clinical practice shows an underprescription of angiotensin converting enzyme inhibitors, both in number of patients and in dosage. The results of the recent ATLAS trial suggest that high doses of lisinopril improve morbidity related to cardiac failure and the combined morbi mortality criterion. The results of this study incite the prescription of high rather than low doses of angiotensin converting enzyme inhibitor. PMID- 10830084 TI - [Digitalis and beta-blockers: what treatment, what dose and for which patient?]. AB - The aim of this update is to review the role of digitalis and betablockers in the treatment of chronic cardiac failure. The role of digitalis is unquestioned in the treatment of chronic cardiac failure complicated by atrial fibrillation resistant to attempts at reduction. Digoxin, with a shorter half-life, is preferred to digitalis. When in sinus rhythm, digoxin is indicated in association with diuretics and angiotensin converting enzyme inhibitors, to improve the quality of life without demonstrable effects on survival. Betablockers now have a recognised place in the treatment of chronic cardiac failure in association with diuretics and angiotensin converting enzyme inhibitors. The compilation of the US Carvedilol HF Trial have shown a 67% reduction in mortality at 6 months. Carvedilol has been officially approved for use in this indication. Some betablockers are under evaluation, regarding positive results in term of mortality, some clinical trials have been stopped. This review emphasises the changes in prescribing habits of digitalis, and mostly betablockers, in chronic cardiac failure. PMID- 10830085 TI - [Cardiac insufficiency: prevention of ventricular arrhythmias]. AB - Ventricular arrhythmias are particularly common in cardiac failure and their mechanisms are very complex. The prevention of these ventricular arrhythmias is only worthwhile if it results in benefits in terms of reduction of the risk of sudden death and in improvement in life expectancy. However, the relationship between complex ventricular arrhythmias and sudden death is far from established. The first problem is, therefore, to select the patients at high risk of sudden death. Unfortunately, there are no reliable markers of arrhythmic risk; only patients at low risk can be reasonably well identified on clinical and haemodynamic assessment and the results of ambulatory and signal averaged ECG. When an antiarrhythmic treatment seems to be required, the choice is very limited in practice. There is no role for Class I antiarrhythmics to play in this indication. Amiodarone, with its complex electrophysiological profile enabling an interaction with all potential mechanisms of ventricular arrhythmias, is a first line drug in cardiac failure because of its efficacy and good myocardial tolerance. However, the benefits of amiodarone therapy in terms of reduction of global mortality have not been demonstrated, especially in view of the discordance between the results of the GESICA and CHF STAT trials. On the other hand, the value of betablockers, whether conventional molecules like bisoprolol (CIBIS II study) or metoprolol (MERIT-HF study), or molecules with a special profile such as carvedilol, has been clearly established. In association with conventional diuretics and angiotensin converting enzyme inhibitors, they reduce global mortality by about 35% and sudden death by 40%. However, the future possibly lies with non-pharmacological approaches such as the implantable defibrillator, at least in patients clearly identified as being at high risk of arrhythmic death, resuscitated from cardiorespiratory arrest due to documented ventricular fibrillation or presenting with haemodynamically poorly tolerated ventricular tachycardia. The automatic defibrillator could improve the prognosis of these patients, irrespective of their functional status (NYHA, Classes I, II or III). In practice, "rhythmological" management of cardiac failure cannot be dissociated from the haemodynamic and neuro-hormonal aspects of the affection, and only a multi-factorial approach is being realistic. PMID- 10830086 TI - [Anticoagulant treatment and cardiac insufficiency]. AB - In cardiac failure, should conventional therapy be associated systematically with anticoagulant or antiplatelet therapy? Embolic complications are uncommon (1 to 2.5% per year) and the benefit/risk ratio seems to be marginal. The absence of prospective randomised controlled trials makes it impossible to give a definitive reply to this question. The indications of oral anticoagulants are based on experience, good sense, the recognition of known embolic risk factors: severe cardiac failure, atrial fibrillation, EF < 0.30 and low VO2 max, mitral valve disease or prosthetic valve, detection of intracavitary thrombus or spontaneous contrast on transoesophageal echocardiography. Aspirin does not seem to be mandatory even if it reduces the thromboembolic risk non-significantly. In this elderly population with a high co-morbidity, the risks of haemorrhage cannot be ignored, and, if oral anticoagulants are prescribed, biological surveillance must be intensive. PMID- 10830087 TI - [Rhythmology Group from the SFC. A 30 year historic evocation]. PMID- 10830088 TI - [Prognostic evaluation of silent ischemia during exercise in patient with recent infarction. Italian multicenter study. SMISS Group]. AB - AIM: The aim of the Multicenter Silent Ischemia Study (SMISS), co-ordinated by the Italian Working Group on Cardiac Rehabilitation, was to evaluate prospectively, the prognostic significance of silent myocardial ischemia during exercise testing in patients with proven ischemic cardiac disease. METHODS: Over a period of six months 4389 consecutive patients performing a maximal symptom limited exercise testing, after drug withdrawal, were enrolled in the 73 ergometric laboratories. All patients were followed up after 12 months, at which time electrocardiogram, examination and clinical history were reassessed. Here we report the results of 1111 patients group with the recent myocardial infarction (inferior 3 months). The follow-up was completed in 1031 (93%) patients. RESULTS: The results of exercise testing were normal in 666 (64.6%) patients; angina alone in 33 (3.2%) patients; silent ischemia in 234 (22.7%) patients; symptomatic ischemia in 98 (9.5%) patients. In 270 patients (26.1%) new events occurred: angina (19.7%); myocardial infarction (3.1%; PTCA (4%); CABG (6%); cardiac death (1.4%). The total events were more common in the patients with exercise induced angina (48.5%) and in those who had exercise induced-symptomatic ischemia (48%), in respect of patients with silent ischemia (29.5%) and of those who had normal testing (20.7%) (p = 0.0001). Myocardial infarction rate was higher in patients with symptomatic ischemia (7.1%) that for those of all other groups (silent ischemia: 1.3%, angina: 3%, normal 3.2%) (p = 0.05). Moreover, the patients with symptomatic ischemia had higher incidence of CABG (p = 0.0001). The mortality rate was low among all patients and did not show differences among the groups. Only among the 31 patients (3%) with blood pressure fall was mortality higher that in patients with a normal blood pressure increase. By multivariate logistic analysis the angina induced by exercise maintained its prognostic significance for all the events, but also other variables were significant: poor exercise tolerance and, between clinical variables angina before myocardial infarction. CONCLUSION: The results showed, in patients who underwent to exercise testing after drug withdrawal, a low incidence of cardiac death and of myocardial infarction on 12 month follow-up; the patients with induced-exercise symptomatic schema had a greater risk for all cardiac events, except for death. PMID- 10830089 TI - [External ventricular support in primary cardiogenic shock]. AB - Primary cardiogenic shock is a common condition with a high mortality rate. In this indication, mechanical assist plays an important part and has improved a lot over the last decade. The authors report their experience with the same assist device in patients with primary cardiogenic shock. Nineteen patients (9 dilated cardiomyopathies, 7 myocardial infarctions, 2 myocardities, 1 undetermined) were treated with an external mechanical ventricular assist device (Thoratec, Berkeley, U.S.). Fourteen patients received a biventricular assist and 5 had a uni-left ventricular assist device. Four of the 19 patients were completely weaned off their ventricular assist after 13, 27, 36 and 94 days, respectively. Ten patients underwent transplantation after an average of 43 days (range 8-95 days). Of the 19 patients, 7 had a portable console allowing autonomous ambulation. Five patients died under mechanical assistance (26.9%) and 3 patients died after transplantation. Three patients required temporary haemodialysis; 4 suffered embolic complications; 4 had mediastinal haemorrhages; 4 had bleeding from other sites, and 6 suffered from late tamponnade. Fourteen patients had at least one infectious episode. The authors conclude that, in patients referred for severe primary cardiogenic shock, the implantation of an external biventricular assist is a reliable option, allowing sequential weaning or being a bridge to transplantation in non-dependent patients, providing they are severely selected. PMID- 10830090 TI - [Transesophageal electrophysiology study, a simple method of investigation of unexplained illness in elderly patients]. AB - Cardiac arrhythmias are common causes of syncope and malaise in elderly patients, but they are sometimes difficult to demonstrate without invasive procedures. The aim of this report was to demonstrate the value of transoesophageal electrophysiological investigation in cases of negative classical non-invasive studies. The authors report 18 cases of patients, aged 70 to 88, mainly in poor general condition, who were admitted for the investigation of malaise or syncope. The ECG was normal or subnormal and Holter monitoring non-contributive to the diagnosis. Transoesophageal electrophysiological study enabled initiation of an arrhythmia, bradycardia or tachycardia, which reproduced the symptoms of spontaneous malaise. In 10 cases, paroxysmal junctional tachycardia was demonstrated, by nodal reentry in 8 cases, and in a latent Kent bundle in 2 cases. The malaise was caused by tachyarrhythmia in 3 patients and by a vagal reaction or sinus arrest after the tachycardia in the other patients. In 6 other patients, atrial fibrillation reproduced the malaise either due to the rapid rhythm, or to bradycardia after the arrhythmia in 3 cases. In another 2 patients, conduction defects were demonstrated by atrial stimulation (alternating bundle branch block in one patient, complete atrioventricular block at the end of atrial stimulation in another patient). The authors conclude that transoesophageal electrophysiological study is a simple technique which allows diagnosis of unexplained malaise in elderly patients when non-invasive methods are unable to demonstrate the causal arrhythmia. Supraventricular arrhythmias seem to be a common and probably underestimated cause of malaise or syncope in elderly patients. PMID- 10830091 TI - [Recommendations of the French Cardiac Society concerning the education of physicians performing coronarography and angioplasty, organization and equipment of coronarography and coronary angioplasty centers]. PMID- 10830092 TI - [Report on interventional coronary cardiology in France. Angiography and Interventional Cardiology Group of the French Society of Cardiology]. PMID- 10830093 TI - [Aspirin, angiotensin converting enzyme inhibitors and cardiac insufficiency]. AB - The possible negative therapeutic interaction between aspirin and angiotensin converting enzyme inhibitors arose from the conclusions of several experimental studies and retrospective analysis of large scale mortality trials with converting enzyme inhibitors. Some experimental results show inhibition of the vasodilatation of converting enzyme inhibitors, increase in pulmonary pressures, vascular resistances and blood pressure, and degradation of renal function and exercise capacity. However, other studies did not confirm these results. In large scale therapeutic trials, some retrospective analyses, but not all of them, have shown less benefit on morbi-mortality of converting enzyme inhibitors in patients on aspirin. The differences between the doses of aspirin, the type and dosage of the converting enzyme inhibitors and neuro-hormonal activation of the patients could explain the discordant results. The results of randomised trials are awaited but, in the meantime, it is logical to propose small doses of aspirin (< or = 100 mg/day) for patients with cardiac failure and atherosclerosis and to avoid the association in all the other patients. PMID- 10830094 TI - [Role of membrane lipids in myocardial cytoprotection]. AB - The cardiomyocyte capacity to regulate ATP production to face any change in energy demand is a major determinant of cardiac function. This process is based on a balanced fatty acid (FA) metabolism, because FA is the main fuel of the heart, although the most expensive one in oxygen. The pathway is, however, weakly controlled by the cardiac myocyte which can well regulate FA mitochondrial entry but not cell FA uptake. For this reason, several pathological situations often result from either harmful accumulation of FA and derivatives or excess FA oxidation. Control of the FA/glucose balance by decreased energy production from FA would thus offer an alternative strategy in the treatment of ischaemia, providing the cardiomyocytes weak ability in handling the non-metabolised FA is controlled. The initiation and the regulation of cardiac contraction both result from membrane activity; the other major role of lipids in the heart is their contribution to membrane homeostasis through phospholipid synthesis pathways and phospholipases. The anti-anginal activity of Trimetazidine, reported as a cytoprotective effect without a haemo-dynamic component; is associated with reduced use of FA for energy. However, accumulation of FA and derivatives has never been observed. Trimetazidine is reported to increase significantly the synthesis of phospholipids without influencing the other lipid classes, thus increasing the incorporation of FA in membrane structures. This cytoprotection appears to be based on the redirection of the use of FA to phospholipid synthesis, which would decrease their availability for energy production. This class of compounds, with the same properties as Trimetazidine, offers a metabolic approach to the treatment of ischaemia. PMID- 10830095 TI - [Aortic dissection at 6 months gestation in a women with Marfan's syndrome. Simultaneous Bentall intervention and cesarean section]. AB - A 31 year old woman with Marfan's syndrome had a dilatation of the aortic root (55-60 mm at the beginning of pregnancy). Pregnancy was continued with beta blocker therapy and with regular echocardiographic follow-up. The aortic dilatation increased (62-65 mm) at the last control and, at the 34th week of pregnancy, the patient suffered a dissection of the ascending aorta. A caesarean section was performed with a Bentall procedure during the same operative session. The mother and baby girl are well two years later. The problems of pregnancy in patients with Marfan's syndrome are discussed. PMID- 10830096 TI - [Right atrial intracardiac varices. Review of the literature and case report]. AB - Right atrial varices are rare. They were described for the first time by anatomo pathologists at the end of the 19th century and beginning of the 20th century. They are situated in the lower part of the inter-atrial septum and rarely exceed 2 cm in diameter. Descriptions have been from post-mortem studies which have led to epidemiological analyses and have given rise to nosological controversies. The authors report a case characterised by the exceptional volume of the varices. This could have enabled the diagnosis to be suspected at transoesophageal echocardiography before surgery. Thoracic CT scan and MRI completed the iconography. In the literature, two other cases of cardiac varices diagnosed at echocardiography have been published: they were small tumours on the lower part of the interatrial septum and the diagnosis before surgery was that of a myxoma. These formations seem to correspond to chance findings and do not appear to give rise to symptoms. PMID- 10830097 TI - [Thoracic aortic aneurysm complicating pseudo-coarctation]. AB - INTRODUCTION: The aneurysm of the descending aorta complicating a pseudocoarctation, itself due to a congenital elongation with kinking of the aorta is a rare entity. OBSERVATION: We report a case of aortic aneurysm discovered in a 72 years old woman without notable antecedents, which was referred for recurrent bronchitis. The X-ray showed a calcified opacity of the upper mediastinum, 5 cm of large. A thoracic CT-scan evoked the presence of a circulating sacciform aneurysm with calcified walls, developing on the final part of the aortic arch, which was with abnormally ascending way going up to the cervico-thoracic orifice and carrying out an aspect of aortic kinking. The assessment was complemented by a RMI as well as an aortic opacification. A thoracic scintigraphy showed an hypoperfusion of the left lung. The remainder of the cardiac assessment was normal. The patient was operated under femoro-femoral extracorporeal circulation through a left posterolateral thoracotomy of the 4th intercostal space. The examination showed a 7 cm diameter calcified aneurysm of the descending thoracic aorta complicating a tight stenosis in connection with an elongation and a kinking. The upper section of the aorta was shifted towards the pleural dome. The aortic section above aneurism was of normal size whereas the lower section was dilated. The aneurism was excised and a prosthetic graft was carried out. The surgery follow-up was marked by an hemodynamic stability, without neurological deficit. A ventilatory assistance was necessary during 5 days. Currently with 8 months follow-up, the patient goes well. COMMENTS: A prosthetic replacement in front of this type of aneurism is legitimate given the risk of the occurrence of complications secondary to the pseudocoarctation (arterial hypertension, aortic insufficiency) or to the aneurism itself, dissection or compression of vicinity (pulmonary artery). PMID- 10830098 TI - [Primitive leiomyosarcoma of the right atrium presenting with a pulmonary embolism]. AB - Primary Cardiac leiomyosarcoma is a very rare condition. The authors report the case of a 64 year old woman admitted to hospital for suspected pulmonary embolism, confirmed by thoracic angio-CT scan. Despite thrombolytic therapy, a persistent poor haemodynamic condition associated with a superior vena cava syndrome led to a transoesophageal echocardiographic examination which demonstrated an abnormal intra-right atrial mass obstructing the tricuspid valve. At surgery, a leio-myosarcoma was successfully resected under good conditions. During systematic three-monthly clinical and echocardiographic follow-up, transoesophageal echocardiography detected an asymptomatic local recurrence at 15 months, for which surgery was repeated with adjuvant chemotherapy. PMID- 10830099 TI - Colworth Medal Lecture. Structural studies of reversible protein phosphorylation and protein phosphatases. PMID- 10830100 TI - Colworth Medal Lecture. Enzymes in the quantum world. AB - Studies on those enzymes and electron-transfer proteins involved in the catabolism of 'C1' substrates in methylotrophic bacteria have provided a wealth of information concerning the transfer of electrons and hydrogen by quantum tunnelling mechanisms. With regard to H-transfer, studies with MADH have provided the first example of ground-state tunnelling of hydrogen driven by the natural, thermally activated, low-frequency motions of the enzyme molecule. Subsequent studies with related enzymes (e.g. TMADH and bacterial sarcosine oxidase) and with thermophilic alcohol dehydrogenase suggest that vibrationally assisted tunnelling of hydrogen may be more widespread than originally assumed. Our studies of electron transfer in TMADH and ETF have established a role for large scale protein dynamics in interprotein electron transfer, and have made a contribution to the ongoing debate concerning the mechanism of amine oxidation by enzymes. Moreover, our work has identified a hitherto unknown mechanism for the control of electron density in reduced flavin that influences the rate of electron transfer between redox centres within a protein molecule. Despite this progress, however, many questions still remain to be resolved. With the development of more sophisticated experimental techniques (and also continued financial support from the funding agencies!), the mechanistic uncertainties surrounding the quantum mechanical transfer of electrons and hydrogen in biological molecules should be transmogrified into the certainties one more readily acquaints with the classical world. PMID- 10830101 TI - MHC linked genes associated with apoptosis/programmed cell death. PMID- 10830102 TI - Bcl-2-related proteins and cancer. PMID- 10830103 TI - c-Jun and Bax: regulators of programmed cell death in developing neurons. PMID- 10830104 TI - Caspases and commitment to cell death. PMID- 10830105 TI - Apoptosis and the regulation of neutrophil lifespan. PMID- 10830106 TI - Epstein-Barr virus and apoptosis: viral mimicry of cellular pathways. PMID- 10830107 TI - Death receptors, NF-kappa B activation and apoptosis: the potential for therapeutic intervention. PMID- 10830108 TI - Structural and functional modelling of von Willebrand factor type A domains in complement and coagulation. PMID- 10830109 TI - Functional role of A-domains in type VI collagen. PMID- 10830110 TI - The matrilins: a growing family of A-domain-containing proteins. PMID- 10830111 TI - Integrin I domains and their function. PMID- 10830112 TI - VIT-1: the second member of a new branch of the von Willebrand factor A domain superfamily. PMID- 10830113 TI - Evolution of von Willebrand factor A (VWA) domains. PMID- 10830114 TI - Lentivirus vectors for gene therapy. PMID- 10830115 TI - Herpes virus vectors for gene therapy in the nervous system. PMID- 10830116 TI - Targeted transfection of neuronal cells using a poly(D-lysine)-cholera-toxin b chain conjugate. PMID- 10830117 TI - Cell-type specific expression in the pituitary: physiology and gene therapy. PMID- 10830118 TI - Adenovirus-mediated cytokine gene transfer in heart allograft transplantation. PMID- 10830119 TI - Immunotherapy of auto-immune diseases by gene transfer. PMID- 10830120 TI - The basic science of brain-tumour gene therapy. PMID- 10830121 TI - Membrane-protein biosynthesis at the endoplasmic reticulum. PMID- 10830122 TI - Expression and purification of the mitochondrial uncoupling proteins: a comparative study between Escherichia coli and Saccharomyces cerevisiae. PMID- 10830123 TI - Expression of prokaryotic membrane transport proteins in Escherichia coli. PMID- 10830124 TI - Improved purification of a rat neurotensin receptor expressed in Escherichia coli. PMID- 10830125 TI - Overexpression and refolding of an 80-kDa iron transporter from the outer membrane of Escherichia coli. PMID- 10830126 TI - Refolding of G-protein-coupled receptors from inclusion bodies produced in Escherichia coli. PMID- 10830127 TI - Amplified expression and membrane reconstitution of transport proteins. PMID- 10830128 TI - Heterologous expression of the red-cell anion exchanger (band 3; AE1). PMID- 10830129 TI - Overexpression of eukaryotic membrane proteins in transgenic tobacco: pioneering the 'green expression system' with the purification and crystallization of recombinant light-harvesting complex II. PMID- 10830130 TI - The use of baculovirus vectors for the production of membrane proteins in insect cells. PMID- 10830131 TI - Molecular chaperones improve functional expression of the serotonin (5 hydroxytryptamine) transporter in insect cells. PMID- 10830132 TI - Large-scale functional expression of visual pigments: towards high-resolution structural and mechanistic insight. PMID- 10830133 TI - Heterologous expression of mammalian and insect neuronal nicotinic acetylcholine receptors in cultured cell lines. PMID- 10830134 TI - Expression and purification of rhodopsin and its mutants from stable mammalian cell lines: application to NMR studies. PMID- 10830135 TI - Transient-expression technologies, their application and scale-up: 5-HT3 serotonin receptor case study. PMID- 10830136 TI - In vitro binding and partitioning of irinotecan (CPT-11) and its metabolite, SN 38, in human blood. AB - The binding of CPT-11 and SN-38 to human plasma proteins was studied by ultrafiltration at 37 degrees C and pH 7.4. In plasma, CPT-11 was 66-60% bound in the range 100-4000 ng/ml and SN-38 was 94-96% bound in the range 50-200 ng/ml. At these concentrations the plasma binding of CPT-11 was slightly saturable, but the plasma binding of SN-38 was concentration-independent. Albumin was the main carrier of CPT-11 and SN-38 in plasma. In blood, the binding of CPT-11 was moderate (80%), mainly to plasma proteins (47%) and erythrocytes (33%). The binding of SN-38 was high (99%) and most of SN-38 in blood was located in blood cells (approximately 66%) The simulation of a grade 3 hematotoxicity (according to National Cancer Institute's Common Toxicity Criteria grading) on the SN-38 blood distribution yielded an increase in fu (free fraction of drug in plasma) from 1.05 to 2.08 and a decrease in C(Bl)/C(P) from 1.66 to 1.14 (both resulting from a decreased cell binding). PMID- 10830137 TI - Development of target-based antineoplastic agents. AB - The elucidation of multiple potential targets in cancer cells and the development of multiple target-based antineoplastic agents provide unique challenges in clinical trial design. Many of these agents are predicted to have cytostatic as opposed to cytotoxic effects and thus the traditional surrogate endpoint of radiologic tumor shrinkage may be inadequate. The ethical and safety issues of obtaining multiple tumor biopsies further complicate the assessment of appropriate target inhibition in patients. We discuss specific issues that need to be addressed during preclinical, phase I, II, and III testing of these agents. We propose clinical trial designs, including a randomized discontinuation design during phase II evaluation, that may be particularly useful for cytostatic antineoplastic agents. PMID- 10830138 TI - Advanced non-small cell lung carcinoma: the emerging role of docetaxel. AB - The treatment of advanced non-small cell lung carcinoma (NSCLC) has improved greatly over the past decade. With the advent of new agents, in particular taxanes, gemcitabine, vinorelbine, and topoisomerase I inhibitors, response rates have improved from 15-20% to 25-35%, with commensurate improvement in median and one year survival rates to 8-10 months and 35-45%, respectively. These improvements have proven statistically significant in multiple studies [1-4]. Docetaxel, either alone or in combination with platinols, has shown particular promise; and, in some arenas, it has become a standard component of our therapeutic armamentarium. We will review the preclinical data and single agent activity of docetaxel in treatment-naive and previously treated NSCLC patients, its activity in combination with cisplatin and carboplatin, as well as other new agents, and finally focus on ongoing studies evaluating its role in locally advanced disease. PMID- 10830139 TI - Gemcitabine in non-small cell lung cancer (NSCLC). AB - The role of chemotherapy in the treatment of non-small cell lung cancer (NSCLC) has increased greatly in the past few years. While cytotoxic drugs are currently used both as single agents and in combination for palliation in locally advanced and metastatic disease, they have also been incorporated into multi-modality treatment strategies of Stage I to Stage III NSCLC. One of the main reasons for the increased acceptance of chemotherapy is the development of new substances. Among the most promising of these new drugs is the antimetabolite gemcitabine. Several single-arm gemcitabine Phase II studies involving more than 400 patients show validated response rates in more than 20% of the patients. These positive results have also been confirmed in randomized Phase II studies. Gemcitabine's unique mechanism of action, its lack of overlapping toxicity with other agents, and its favorable toxicity profile also define it as an ideal candidate for combination therapy. The activity seen with single-agent gemcitabine therapy can be compared with that of cisplatin-etoposide combination therapy. Gemcitabine cisplatin combination response rates range from 31% to 54%, with a median survival time between 8.4 and 15.4 months and a 1-year survival rate between 30% and 59%. In addition to the clinical research of gemcitabine-cisplatin combinations, gemcitabine has also been tested in various double and triple combinations with carboplatin, paclitaxel, docetaxel, vinorelbine, and ifosfamide. Investigations combining gemcitabine with radiation therapy are on going. The following review will summarize results from representative Phase I/II and III studies using gemcitabine for NSCLC patients. PMID- 10830140 TI - Current approaches and future strategies for pancreatic carcinoma. AB - Pancreatic cancer is a lethal disease characterized by local invasion and early dissemination. It is resistant to conventional surgical, radiotherapeutic, and chemotherapeutic modalities. These interventions have had minimal impact on overall survival with very few patients enjoying long term survival. Over the past few years, 2'difluoro-2'deoxycytidine (gemcitabine) has demonstrated modest activity in this disease and investigations are proceeding to expand its role in combination with radiotherapy and other chemotherapeutic agents. In addition, the identification of the molecular defects underlying this disease has suggested molecular targets for the design of rational systemic therapy. These targets include matrix metalloproteinases, K-ras, HER2/neu, p53, and the epidermal growth factor receptor. Current and future clinical trials designed to improve the survival of patients with pancreatic cancer will be discussed. PMID- 10830143 TI - The 7 habits of highly effective psychopharmacologists: overview. PMID- 10830142 TI - A phase I trial of cisplatin plus decitabine, a new DNA-hypomethylating agent, in patients with advanced solid tumors and a follow-up early phase II evaluation in patients with inoperable non-small cell lung cancer. AB - The authors describe a phase I trial of cisplatin plus decitabine, a novel DNA hypomethylating agent, in patients with advanced solid tumors, which was followed by an early phase II evaluation of the combination in patients with inoperable non-small cell lung cancer (NSCLC). In the phase I trial, cisplatin was studied at a fixed dose of 33 mg/m2, while decitabine was escalated in four (I-IV) dose escalation levels (45, 67, 90 to 120 mg/m2, respectively) in consecutive groups of at least 3 patients per dose level. Decytabine was administered to the patients as a two-hour intravenous infusion, while cisplatin was given intravenously immediately after the end of decitabine infusion. Both agents were given on days 1-3 every 21 days. Twenty-one patients were included in the phase I trial. Dose level IV (120 mg/m2 decitabine) was considered the maximum tolerated dose (MTD), while the dose-limiting toxicities were neutropenia, thrombocytopenia and mucositis. The recommended doses for phase II trials in good- and poor-risk patients were 90 (level III) and 67 mg/m2 (level II), respectively. One short lasting partial response was observed in a patient with cervical cancer, while two minor regression were documented in a patients with NSCLC and cervical cancer, respectively. Dose level II was selected for the phase II trial in patients with inoperable NSCLC. Fourteen consecutive patients were included in this part of the study. The median age of the patients was 57 years (range, 39 75), male/female ratio of 11/3 and a median WHO performance status 1 (0-2). The stage of disease were IIIB (5) and IV (9). Prior irradiation to the chest was given in one case. A total of 30 treatment courses were evaluable for toxicity and response, with a median of 2 courses per patient (1-4). Grade 3-4 neutropenia and thrombocytopenia were observed in about half of the cases. Mucositis, diarrhea, nausea and vomiting, and skin rash were also observed in some patients. Three minor responses were documented, which lasted for 4, 16 and 36 weeks. Median survival of patients was 15 weeks (4-38). In conclusion, the cisplatin plus decitabine combination did not exhibit significant antitumor activity in patients with NSCLC at the dose and schedule applied in this trial to justify its further evaluation in this patient population. PMID- 10830144 TI - Prevalence of attention-deficit/hyperactivity disorder and conduct disorder among substance abusers. AB - BACKGROUND: This cross-sectional study sought to determine the prevalence of attention-deficit/hyperactivity disorder (ADHD) and conduct disorder among adults admitted to 2 chemical dependency treatment centers. It was hypothesized that ADHD alone or in combination with conduct disorder would be overrepresented in a population of patients with psychoactive substance use disorders. METHOD: Two hundred one participants were selected randomly from 2 chemical dependency treatment centers. Standardized clinical interviews were conducted using the Structured Clinical Interview for DSM-IV, the Addiction Severity Index, and DSM IV criteria for ADHD. Reliabilities for the diagnostic categories were established using the Cohen kappa, and the subgroups of individuals with and without ADHD and conduct disorder were compared. RESULTS: Forty-eight (24%) of the participants were found to meet DSM-IV criteria for ADHD. The prevalence of ADHD was 28% in men (30/106) and 19% in women (18/95; NS). Seventy-nine participants (39%) met criteria for conduct disorder, and 34 of these individuals also had ADHD. Overall, individuals with ADHD (compared with those without ADHD) were more likely to have had more motor vehicle accidents. Women with ADHD (in comparison with women without ADHD) had a higher number of treatments for alcohol abuse. Individuals with conduct disorder (in comparison with those without conduct disorder) were younger, had a greater number of jobs as adults, and were more likely to repeat a grade in school, have a learning disability, be suspended or expelled from school, have an earlier age at onset of alcohol dependence, and have had a greater number of treatments for drug abuse. They were more likely to have a lifetime history of abuse of and/or dependence on cocaine, stimulants, hallucinogens, and/or cannabis. CONCLUSION: A significant overrepresentation of ADHD exists among inpatients with psychoactive substance use disorders. Over two thirds of those with ADHD in this sample also met criteria for conduct disorder. Our sample had a very large overlap between ADHD and conduct disorder, and the major comorbidities identified here were attributable largely to the presence of conduct disorder. Individuals who manifest conduct disorder and/or ADHD represent a significant proportion of those seeking treatment for psychoactive substance use disorders. They appear to have greater comorbidity and may benefit from a treatment approach that addresses these comorbidities specifically through medical and behavioral therapies. PMID- 10830145 TI - Pharmacokinetics, tolerability, and clinical effectiveness of quetiapine fumarate: an open-label trial in adolescents with psychotic disorders. AB - BACKGROUND: This is the first investigation of the pharmacokinetics, tolerability, and efficacy of quetiapine fumarate in adolescents with chronic or intermittent psychotic disorders. METHOD: Ten patients with DSM-IV chronic or intermittent psychotic disorders (ages 12.3 through 15.9 years) participated in an open-label, rising-dose trial and received oral doses of quetiapine twice daily (b.i.d.), starting at 25 mg b.i.d. and reaching 400 mg b.i.d. by day 20. The trial ended on day 23. Key assessments were pharmacokinetic analysis of plasma quetiapine concentrations and neurologic, safety, and efficacy evaluations. RESULTS: No statistically significant differences were observed between 100-mg b.i.d. and 400-mg b.i.d. quetiapine regimens for total body clearance, dose-normalized area under the plasma concentration-time curve, or dose-normalized premorning- or postmorning-dose trough plasma values obtained under steady-state conditions after multiple-dose regimens. No unexpected side effects occurred with quetiapine therapy, and no statistically significant changes from baseline were observed for the UKU Side Effect Rating Scale items that were rated. No serious adverse events or clinically important changes in hematology or clinical chemistry variables were reported. The most common adverse events were postural tachycardia and insomnia. Extrapyramidal side effects improved, as evidenced by significant (p < .05) decreases from baseline to endpoint in the mean Simpson-Angus Scale total scores and Barnes Akathisia Scale scores. Quetiapine improved positive and negative symptoms, as shown by significant (p < .05) decreases from baseline to endpoint in the mean Brief Psychiatric Rating Scale total score, the Clinical Global Impressions-Severity of Illness scale, and the Modified Scale for the Assessment of Negative Symptoms summary score. CONCLUSION: Quetiapine pharmacokinetics were dose proportional in adolescents and were similar to those previously reported for adults. Quetiapine was well tolerated and effective in the small number of adolescents studied. PMID- 10830146 TI - Recurrent cases of corticosteroid-induced mood disorder: clinical characteristics and treatment. AB - BACKGROUND: Corticosteroids often induce steroid psychosis, a collection of heterogeneous syndromes with different pathophysiologic mechanisms. To date, no study has focused specifically on recurrent corticosteroid-induced mood disorders and considered their long-term outcome and treatment strategies. METHOD: Nine patients whose initial clinical presentation met DSM-IV criteria for a substance induced mood disorder were identified by a review of medical records. Their clinical characteristics and treatments were examined. RESULTS: All 9 corticosteroid-treated patients had a clinical course of bipolar disorder. Seven patients initially developed a manic or hypomanic state with subacute onset ranging from 1 to 3 months. Six patients had manic episodes accompanied by psychotic features. The proportion of manic episodes relative to total mood episodes of the 9 patients was 65.6%, suggesting manic predominance. Seven patients showed mood episodes that had no direct relationship to corticosteroid therapy and were preceded by various psychosocial stressors. Four of 5 patients who received steroid pulse therapy rapidly became manic or hypomanic. Antidepressants as well as mood stabilizers were useful for treatment of the present 9 patients. CONCLUSION: Recurrent cases of corticosteroid-induced mood disorder have interesting clinical features, such as subacute onset, manic predominance, frequent accompanying psychotic features, and similar recurrent episodes in association with psychosocial stressors and corticosteroid use. Management, including psychopharmacologic intervention, should be indicated by a consideration of the underlying illnesses and psychosocial stressors. PMID- 10830141 TI - Promising approaches in acute leukemia. AB - In the last few decades, there has been a significant improvement in the prognosis of patients with acute leukemias. Still, the majority of patients succumb to these diseases. In recent years there has been a great surge in the understanding of the molecular mechanisms of disease which have provided us with new targets for anti-leukemia therapy. These range from chemotherapeutic agents with novel mechanisms of action, such as topoisomerase I inhibitors or demethylating agents, to reversal of drug-resistance mechanisms, to monoclonal antibodies directed against specific antigens, and targeted therapy that inhibit the function of molecules such as tyrosine kinases or Ras. The research on many of these agents is still in the early phases, but these new approaches offer the promise of finding a cure for the majority of patients with leukemia in the near future. Here we describe some of the promising approaches that are currently being investigated in the treatment of acute leukemias. PMID- 10830147 TI - Social functioning in depression: a review. AB - OBJECTIVE: This article reviews the available data on social functioning in depression and provides clinical guidelines and opinion on this important and expanding field. DATA SOURCES: A MEDLINE search was conducted to identify all English-language articles (1988-1999) using the search terms depression and social functioning, depression and social adjustment, depression and psychosocial functioning, and social functioning and antidepressant. Further articles were obtained from the bibliographies of relevant articles. DATA SYNTHESIS: Depressive disorders are frequently associated with significant and pervasive impairments in social functioning, often substantially worse than those experienced by patients with other chronic medical conditions. The enormous personal, social, and economic impact of depression, due in no small part to the associated impairments in social functioning, is often underappreciated. Both pharmacologic and psychotherapeutic approaches can improve social impairments, although there is a lack of extended, randomized controlled trials in this area using consistent assessment criteria. CONCLUSION: Despite this lack, it is becoming clear that not all treatments are equally effective in relieving the impaired social functioning associated with depressive disorders. Furthermore, efficacy in relieving the core symptoms of depression does not necessarily guarantee efficacy in relieving impaired social functioning. PMID- 10830149 TI - Nefazodone and the treatment of nonparaphilic compulsive sexual behavior: a retrospective study. AB - BACKGROUND: Recent reports suggest that individuals with nonparaphilic compulsive sexual behavior can be treated pharmacologically with selective serotonin reuptake inhibitors (SSRIs) to control sexual obsessions and compulsions. However, these medications have produced sexual side effects that may limit long term use, particularly as individuals strive to reestablish healthy sexual relationships. Nefazodone is an antidepressant that is not associated with the sexual side effects of other SSRIs. We examined retrospective data from our clinic to investigate whether nefazodone has utility in the treatment of nonparaphilic compulsive sexual behavior. METHOD: Fourteen subjects who met DSM IV criteria for sexual disorder NOS as well as criteria used by our research group for nonparaphilic compulsive sexual behavior and who had been treated with nefazodone were selected from patient charts at our clinic. The treating physician abstracted information from the charts regarding comorbid psychiatric conditions, medication, dosage, treatment response, and side effects. RESULTS: In this study, the mean dosage of nefazodone was 200 mg/day. Of the subjects who remained on long-term nefazodone therapy, 6 (55%) reported good control of sexual obsessions and compulsions, and 5 (45%) reported a remission of sexual obsessions and compulsions. CONCLUSION: Results from this preliminary retrospective study suggest that nefazodone decreases the frequency of sexual obsessions and compulsions but does not produce the undesired sexual side effects caused by SSRI treatment. PMID- 10830148 TI - Antidepressant-induced sexual dysfunction during treatment with moclobemide, paroxetine, sertraline, and venlafaxine. AB - BACKGROUND: Recent reports suggest that adverse effects on sexual function occur in up to 50% of patients who are treated with selective serotonin reuptake inhibitor (SSRI) antidepressants. Previously cited low rates were more likely a function of underreporting than underoccurrence. There is less evidence about rates of dysfunction with serotonin-norepinephrine reuptake inhibitor (SNRI) and reversible inhibitor of monoamine oxidase A (RIMA) antidepressants. The purpose of this report is to evaluate disturbances in sexual drive/desire and arousal/orgasm in 107 patients who met criteria for major depressive disorder and received treatment with either moclobemide, paroxetine, sertraline, or venlafaxine. METHOD: All consenting eligible patients who met DSM-IV criteria for major depressive disorder completed the Sexual Functioning Questionnaire, version 1 (SFQ) and were assessed using the 17-item Hamilton Rating Scale for Depression (HAM-D) prior to and after 8 or 14 weeks of antidepressant therapy. Analyses were carried out to examine the effect of gender, drug type, pretreatment level of sexual dysfunction, and drug response on reported sexual dysfunction. RESULTS: Compared with women, men experienced a significantly greater level of drug related impairment in drive/desire (p < .05), whereas there were no statistically significant differences in levels of arousal/orgasm impairment between men and women. The reported impairment in drive/desire items for men ranged from 38% to 50% and from 26% to 32% for women. No differences were found across the 4 antidepressants in men, whereas in women, rates of dysfunction were generally higher with sertraline and paroxetine, but only significantly so in comparison with moclobemide on some measures (p < .03). Rates of sexual dysfunction with venlafaxine tended to fall between those of SSRIs and the RIMA agent. An unexpected relationship was found between favorable drug response and a decreased level of drug-induced sexual dysfunction. CONCLUSION: Antidepressant-induced sexual dysfunction occurs in approximately 30% to 70% of patients who are treated with sertraline or paroxetine. Lower rates are reported with moclobemide and venlafaxine. Clinicians should evaluate the various aspects of sexual dysfunction before and during antidepressant therapy. PMID- 10830150 TI - Combined treatment with venlafaxine and tricyclic antidepressants in depressed patients who had partial response to clomipramine or imipramine: initial findings. AB - BACKGROUND: We report, after 3 years of work, a case series showing our initial results (efficacy, tolerability, and safety) with the addition of venlafaxine immediate release (IR) to either clomipramine or imipramine in depressed patients who had shown only partial response to maximal doses of one of those tricyclic antidepressants (TCAs) and no further improvement after addition of usual augmentation drugs. METHOD: Eleven patients were treated, 10 of them having a recurrent depressive disorder (DSM-IV) and all of them having current major depression (DSM-IV) that in 9 patients was moderate or severe despite intense TCA treatment as well as usual augmentations. Under open and outpatient conditions, we maintained TCA doses, discontinued previous augmentations, and then added venlafaxine IR to a maximum dosage, if necessary, of 150 mg every 12 hours. There was no control group. Response was assessed using the 17-item Hamilton Rating Scale for Depression (HAM-D), DSM-IV criteria, the Clinical Global Impressions Severity of Illness scale, and persistence of improvements after 6 months. We measured clinical tolerance (using the UKU Side Effect Rating Scale), blood pressure and heart rate, electrocardiogram (ECG), and blood TCA levels after adding venlafaxine IR. RESULTS: A sustained improvement (> 50% decrease in HAM-D score plus decrease in DSM-IV severity level) appeared in 9 patients, and sustained full remission (DSM-IV criteria plus HAM-D score < 5) in 7. Panic agoraphobic symptoms improved in the 2 patients suffering from them. There were no dropouts, and tolerability was good. No significant changes in blood pressure and heart rate, ECG, or blood tricyclic levels were found. CONCLUSION: Addition of venlafaxine to clomipramine or imipramine could be an effective and safe augmentation strategy in depressive patients with partial response to maximum dose monotherapy. A consistent replication of these initial findings is strongly needed. PMID- 10830151 TI - One-year costs of second-line therapies for depression. AB - BACKGROUND: We compared patterns of medical resource utilization and costs among patients receiving a serotonin-norepinephrine reuptake inhibitor (venlafaxine), one of the selective serotonin reuptake inhibitors (SSRIs), one of the tricyclic agents (TCAs), or 1 of 3 other second-line therapies for depression. METHOD: Using claims data from a national managed care organization, we identified patients diagnosed with depression (ICD-9-CM criteria) who received second-line antidepressant therapy between 1993 and 1997. Second-line therapy was defined as a switch from the first class of antidepressant therapy observed in the data set within 1 year of a diagnosis of depression to a different class of antidepressant therapy. Patients with psychiatric comorbidities were excluded. RESULTS: Of 981 patients included in the study, 21% (N = 208) received venlafaxine, 34% (N = 332) received an SSRI, 19% (N = 191) received a TCA, and 25% (N = 250) received other second-line antidepressant therapy. Mean age was 43 years, and 72% of patients were women. Age, prescriber of second-line therapy, and prior 6-month expenditures all differed significantly among the 4 therapy groups. Total, depression-coded, and non-depression-coded 1-year expenditures were, respectively, $6945, $2064, and $4881 for venlafaxine; $7237, $1682, and $5555 for SSRIs; $7925, $1335, and $6590 for TCAs; and $7371, $2222, and $5149 for other antidepressants. In bivariate analyses, compared with TCA-treated patients, venlafaxine- and SSRI-treated patients had significantly higher depression-coded but significantly lower non-depression-coded expenditures. Venlafaxine was associated with significantly higher depression-coded expenditures than SSRIs. However, after adjustment for potential confounding covariables in multivariate analyses, only the difference in depression-coded expenditures between SSRI and TCA therapy remained significant. CONCLUSION: After adjustment for confounding patient characteristics, 1-year medical expenditures were generally similar among patients receiving venlafaxine, SSRIs, TCAs, and other second-line therapies for depression. Observed differences in patient characteristics and unadjusted expenditures raise questions as to how different types of patients are selected to receive alternative second-line therapies for depression. PMID- 10830152 TI - Schizophrenia-associated idiopathic unconjugated hyperbilirubinemia (Gilbert's syndrome): 3 case reports. PMID- 10830153 TI - Effectiveness of risperidone in simple schizophrenia: a single case report. PMID- 10830155 TI - Gastrointestinal adverse effects with divalproex sodium and valproic acid. PMID- 10830154 TI - Citalopram and clozapine: potential drug interaction. PMID- 10830156 TI - Misuse of the term phenomenology. PMID- 10830157 TI - Low-dose lithium augmentation of divalproex in geriatric mania. PMID- 10830158 TI - New insights into genetics and pathophysiology of Alzheimer's disease: what are the clinical and therapeutic implications? PMID- 10830159 TI - Explanatory therapy in hypochondriasis. AB - OBJECTIVE: The aim of the study was to evaluate the effectiveness of individual explanatory therapy in hypochondriasis. METHOD: Twenty patients with DSM-IV hypochondriasis were randomly assigned to 2 groups. One received explanatory therapy and was assessed again after a 6-month follow-up. The other was first assigned to a waiting list and subsequently treated with explanatory therapy. All patients received usual medical care from their physicians. Assessments involved both observer and self-rated instruments. RESULTS: In both groups, explanatory therapy was significantly associated with a reduction of hypochondriacal fears and beliefs, improvement in affective disturbances and anxiety sensitivity, and a decrease in health care utilization. Therapeutic gains were maintained at follow up. Substantial residual symptomatology, however, remained. CONCLUSION: The results suggest that hypochondriasis is a treatable condition and that explanatory therapy is a viable therapeutic tool. Further research should disclose the actual components of the mechanism of change in hypochondriacal patients. PMID- 10830160 TI - A proteasome howdunit: the case of the missing signal. PMID- 10830161 TI - PARsing embryonic polarity. PMID- 10830162 TI - Mimicry grasps reality in translation termination. PMID- 10830163 TI - Eukaryotic fatty acylation drives plasma membrane targeting and enhances function of several type III effector proteins from Pseudomonas syringae. AB - Bacterial pathogens of plants and animals utilize conserved type III delivery systems to traffic effector proteins into host cells. Plant innate immune systems evolved disease resistance (R) genes to recognize some type III effectors, termed avirulence (Avr) proteins. On disease-susceptible (r) plants, Avr proteins can contribute to pathogen virulence. We demonstrate that several type III effectors from Pseudomonas syringae are targeted to the host plasma membrane and that efficient membrane association enhances function. Efficient localization of three Avr proteins requires consensus myristoylation sites, and Avr proteins can be myristoylated inside the host cell. These prokaryotic type III effectors thus utilize a eukaryote-specific posttranslational modification to access the subcellular compartment where they function. PMID- 10830164 TI - PKA phosphorylation dissociates FKBP12.6 from the calcium release channel (ryanodine receptor): defective regulation in failing hearts. AB - The ryanodine receptor (RyR)/calcium release channel on the sarcoplasmic reticulum (SR) is the major source of calcium (Ca2+) required for cardiac muscle excitation-contraction (EC) coupling. The channel is a tetramer comprised of four type 2 RyR polypeptides (RyR2) and four FK506 binding proteins (FKBP12.6). We show that protein kinase A (PKA) phosphorylation of RyR2 dissociates FKBP12.6 and regulates the channel open probability (Po). Using cosedimentation and coimmunoprecipitation we have defined a macromolecular complex comprised of RyR2, FKBP12.6, PKA, the protein phosphatases PP1 and PP2A, and an anchoring protein, mAKAP. In failing human hearts, RyR2 is PKA hyperphosphorylated, resulting in defective channel function due to increased sensitivity to Ca2+-induced activation. PMID- 10830165 TI - The Drosophila homolog of C. elegans PAR-1 organizes the oocyte cytoskeleton and directs oskar mRNA localization to the posterior pole. AB - In C. elegans, the PAR-1 kinase is localized to the posterior of the zygote and is required for anterior-posterior axis formation. Here, we report that a Drosophila PAR-1 homolog localizes to the posterior of the oocyte with oskar mRNA. Furthermore, par-1 mutants show a novel polarity phenotype in which bicoid mRNA accumulates normally at the anterior, but oskar mRNA is redirected to the center of the oocyte, resulting in embryonic patterning defects. These phenotypes arise from a disorganization of the oocyte microtubule cytoskeleton, consistent with reports that mammalian PAR-1 homologs regulate microtubule dynamics. Thus, Drosophila PAR-1 may remodel the oocyte microtubule network to define the posterior as the site for oskar localization. These results identify a molecular parallel between anterior-posterior polarization in Drosophila and C. elegans. PMID- 10830166 TI - Cytochrome c deficiency causes embryonic lethality and attenuates stress-induced apoptosis. AB - Cytochrome c released from mitochondria has been proposed to be an essential component of an apoptotic pathway responsive to DNA damage and other forms of cell stress. Murine embryos devoid of cytochrome c die in utero by midgestation, but cell lines established from early cytochrome c null embryos are viable under conditions that compensate for defective oxidative phosphorylation. As compared to cell lines established from wild-type embryos, cells lacking cytochrome c show reduced caspase-3 activation and are resistant to the proapoptotic effects of UV irradiation, serum withdrawal, or staurosporine. In contrast, cells lacking cytochrome c demonstrate increased sensitivity to cell death signals triggered by TNFalpha. These results define the role of cytochrome c in different apoptotic signaling cascades. PMID- 10830167 TI - Tim23 links the inner and outer mitochondrial membranes. AB - Tim23, a key component of the mitochondrial preprotein translocase, is anchored in the inner membrane by its C-terminal domain and exposes an intermediate domain in the intermembrane space that functions as a presequence receptor. We show that the N-terminal domain of Tim23 is exposed on the surface of the outer membrane. The two-membrane-spanning topology of Tim23 is a novel characteristic in membrane biology. By the simultaneous integration into two membranes, Tim23 forms contacts between the outer and inner mitochondrial membranes. Tethering the inner membrane translocase to the outer membrane facilitates the transfer of precursor proteins from the TOM complex to the TIM23 complex and increases the efficiency of protein import. PMID- 10830168 TI - Crystal structures of two FGF-FGFR complexes reveal the determinants of ligand receptor specificity. AB - To elucidate the structural determinants governing specificity in fibroblast growth factor (FGF) signaling, we have determined the crystal structures of FGF1 and FGF2 complexed with the ligand binding domains (immunoglobulin-like domains 2 [D2] and 3 [D3]) of FGF receptor 1 (FGFR1) and FGFR2, respectively. Highly conserved FGF-D2 and FGF-linker (between D2-D3) interfaces define a general binding site for all FGF-FGFR complexes. Specificity is achieved through interactions between the N-terminal and central regions of FGFs and two loop regions in D3 that are subject to alternative splicing. These structures provide a molecular basis for FGF1 as a universal FGFR ligand and for modulation of FGF FGFR specificity through primary sequence variations and alternative splicing. PMID- 10830169 TI - The crystal structure of the ligand binding module of axonin-1/TAG-1 suggests a zipper mechanism for neural cell adhesion. AB - We have determined the crystal structure of the ligand binding fragment of the neural cell adhesion molecule axonin-1/TAG-1 comprising the first four immunoglobulin (Ig) domains. The overall structure of axonin-1(Ig1-4) is U-shaped due to contacts between domains 1 and 4 and domains 2 and 3. In the crystals, these molecules are aligned in a string with adjacent molecules oriented in an anti-parallel fashion and their C termini perpendicular to the string. This arrangement suggests that cell adhesion by homophilic axonin-1 interaction occurs by the formation of a linear zipper-like array in which the axonin-1 molecules are alternately provided by the two apposed membranes. In accordance with this model, mutations in a loop critical for the formation of the zipper resulted in the loss of the homophilic binding capacity of axonin-1. PMID- 10830170 TI - A homeodomain protein code specifies progenitor cell identity and neuronal fate in the ventral neural tube. AB - Distinct classes of neurons are generated at defined positions in the ventral neural tube in response to a gradient of Sonic Hedgehog (Shh) activity. A set of homeodomain transcription factors expressed by neural progenitors act as intermediaries in Shh-dependent neural patterning. These homeodomain factors fall into two classes: class I proteins are repressed by Shh and class II proteins require Shh signaling for their expression. The profile of class I and class II protein expression defines five progenitor domains, each of which generates a distinct class of postmitotic neurons. Cross-repressive interactions between class I and class II proteins appear to refine and maintain these progenitor domains. The combinatorial expression of three of these proteins--Nkx6.1, Nkx2.2, and Irx3--specifies the identity of three classes of neurons generated in the ventral third of the neural tube. PMID- 10830171 TI - Mixed cryoglobulinemia associated with hepatitis C virus infection. PMID- 10830172 TI - Re-emergence of paragonimiasis. PMID- 10830173 TI - Obstructive nephropathy. AB - Obstructive nephropathy is a relatively common entity that is treatable and often reversible. It occurs at all ages from infancy to elderly subjects. Obstructive uropathy is classified according to the degree, duration and site of the obstruction. It is the result of functional or anatomic lesions located in the urinary tract. The causes of obstructive uropathy are many. Obstruction of the urinary tract may decrease renal blood flow and the glomerular filtration rate. Several abnormalities in tubular function may occur in obstructive nephropathy. These include decreased reabsorption of solutes and water, inability to concentrate the urine and impaired excretion of hydrogen and potassium. Renal interstitial fibrosis is a common finding in patients with long-term obstructive uropathy. Several factors: macrophages, growth factors, hypoxia, cytokines are involved in the pathogenesis of interstitial fibrosis. It has been shown that ACE inhibitors ameliorate the interstitial fibrosis in animals with obstructive uropathy. PMID- 10830174 TI - Possible risk factors of carotid artery atherosclerosis in the Japanese population: a primary prevention study in non-diabetic subjects. AB - OBJECTIVE: Hyperinsulinemia has been associated with the risk of coronary heart disease, stroke, and renal disease in nondiabetic subjects. However, direct evidence that hyperinsulinemia per se is directly associated with atherosclerosis has been conflicting. The present study was designed to investigate the cross sectional association of carotid artery atherosclerosis with insulin, independent of well-known cardiovascular risk factors, in nondiabetic subjects. METHODS AND SUBJECTS: Between 1996 and 1997, 1,335 subjects (620 men and 715 women) were recruited from one Japanese community, interviewed, and examined. Clinical measurements in the study included intimal-medial thickness (IMT) of the carotid artery, fasting plasma insulin, serum total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), fasting plasma glucose (FPG), hemoglobin type HbA1c, systolic blood pressure (SBP), diastolic blood pressure (DBP), and body mass index (BMI). We divided the subjects of both genders into three subgroups according to age (40-49 years of age; 50-59; and 60-69). RESULTS: Using simple regression analysis, we found that IMT was significantly correlated with at least one of TC, LDL-C, HbA1c, SBP, DBP, and BMI in each subgroup. The results of multivariate analysis showed that IMT was independently correlated with TC, HDL-C, LDL-C, SBP and BMI in males and with TC, TG, HDL-C, LDL-C, HbA1c, SBP, DBP, and BMI in females. Insulin levels showed no correlation with IMT in either males or females. CONCLUSION: Fasting hyperinsulinemia does not appear to be correlated with carotid artery atherosclerosis based on the present cross-sectional results. PMID- 10830175 TI - Prognostic significance of electrocardiographic change during anginal attack in patients with unstable angina. AB - OBJECTIVE AND METHODS: We examined the prognostic significance of electrocardiographic change during anginal attack, C-reactive protein and fibrinogen in 169 patients with unstable angina. RESULTS: During the 90-day follow-up period, 26 patients (15%) exhibited new cardiac events (death, myocardial infarction or urgent revascularization). Using multivariate analysis, ST depression (relative risk 7.507 [95 % confidence intervals 1.842-30.592], p<0.01) during anginal attack was found to be an independent risk factor to predict cardiac events as well as diabetes mellitus, an increased total cholesterol level and the use of a thrombolytic agent. C-reactive protein and fibrinogen did not have prognostic significance. CONCLUSION: ST depression during anginal attack is an independent risk predictor for new cardiac events in patients with unstable angina. PMID- 10830176 TI - Age related differences in electroencephalographic and cardiac arousal at the termination of sleep apnea/hypopnea. AB - OBJECTIVE: The purpose of the present study was to determine whether electroencephalographic (EEG) and cardiac arousal, i.e. heart rate elevation at the termination of apnea/hypopnea are related to aging. METHODS: The subjects were 13 middle-aged (40-60 years of age, 47.8+/-5.35 years) and 10 elderly (>60 years of age, 69.8+/-5.31 years) male patients with obstructive sleep apnea syndrome. We evaluated the number of apneas per an hour of sleep (apnea index: AI), the number of hypopneas per an hour of sleep (hypopnea index: HI), the summation of HI and AI (apnea/hypopnea index: AHI), sleep stage, the amount of time during which nocturnal oxygen saturation was decreased below 90% (oxygen desaturation time: ODT), and EEG and cardiac arousal at the termination of apnea/hypopnea using polysomnography with pulse oximetry. RESULTS: There was no significant difference in AHI, duration of apnea/hypopnea, and sleep stage distribution between the two groups. However, the ratio of apnea/hypopnea with EEG and cardiac arousal was significantly higher in middle-aged than in elderly patients. The ratio of HI to AHI was significantly lower in middle-aged than in elderly patients. In middle-aged patients, EEG and cardiac arousal were significantly correlated with AI, AHI, and ODT, whereas, in elderly patients, these parameters were not similarly correlated. CONCLUSION: Our findings suggest that the differences in EEG and cardiac arousal, and the pattern of sleep disordered breathing (SDB) between middle-aged and elderly patients with SDB might be a physiological phenomenon of aging. PMID- 10830177 TI - Effectiveness of the fold plication method in lung volume reduction surgery. AB - OBJECT: The fold plication method is a new operative procedure for lung volume reduction surgery whereby the target area is obliterated by plicating the folded tissue using a knifeless stapler, without the use of bovine pericardium. The effectiveness of this new method was evaluated in patients with advanced pulmonary emphysema. PATIENTS AND METHODS: Two weeks before and 6 months after surgery, pulmonary function, static lung compliance, maximal esophageal pressure, maximal inspiratory and expiratory mouth pressures, 6-min walking distance and the Borg scale were determined in twenty consecutive patients who underwent video assisted thoracoscopic unilateral surgery. RESULTS: There was an increase in forced expiratory volume in one second (31%), forced vital capacity, peak expiratory flow rate and maximal voluntary ventilation, and a decrease in functional residual capacity (-16%) measured by plethysmograph. Static lung compliance decreased, and maximal esophageal pressure, and maximal inspiratory and expiratory mouth pressures increased. The 6-min walking distance increased (20%) and the Borg scale decreased (5.9 to 3.5). CONCLUSION: The results compare favorably with those obtained with other methods. Thus, the fold plication method could be considered an alternative procedure for lung volume reduction surgery. PMID- 10830178 TI - Diffuse angiodysplasia of the upper gastrointestinal tract in a patient with hypertrophic obstructive cardiomyopathy. AB - A 64-year-old woman with a known history of hypertrophic obstructive cardiomyopathy presented with severe anemia of unknown origin. She had also suffered from repeated episodes of upper gastrointestinal bleeding for the previous 3 years. Despite bone marrow examination and panendoscopic and angiographic studies, the origin of anemia remained undefined until a small bleeding site was found during a duodenoscopic examination. The lesion proved to be angiodysplasia. This case report is interesting in that angiodysplasia elicited gastrointestinal bleeding and was the cause of anemia. In the international literature, there are very few reported cases of bleeding from gastrointestinal angiodysplasia in association with subvalvular aortic obstruction. PMID- 10830179 TI - Fourteen-year survival in a case of ventricular septal perforation after myocardial infarction. AB - We treated a case of ventricular septal perforation (VSP) who survived for 14 years after myocardial infarction. Nine years after the onset of myocardial infarction, an apparent cardiac murmur was discovered by chance, and following further examination, the patient was diagnosed as having VSP. The patient is still in the NYHA functional class I, and requiring no surgical treatment. In general, the prognosis of myocardial infarction complicated with VSP is so poor that there have only been 14 reported cases of long survival without surgical treatment. Among these patients, only 2 survived for more than 10 years. The present case is extremely rare, and evidently is the longest living survivor with this condition yet reported. PMID- 10830180 TI - Osteomalacia that became symptomatic 13 years after a total gastrectomy. AB - A 66-year-old man who underwent a total gastrectomy 13 years ago was admitted to our hospital complaining of severe low back pain and muscle weakness. Biochemical examinations revealed hypocalcemia, hypophosphathemia, low serum 25 (OH) vitamin D and hyperparathyroidism. A chest CT scan revealed pseudofractured ribs, whereas plain X-photography did not show any significant findings. We diagnosed the illness as osteomalacia due to malabsorption. The patient has been receiving oral active vitamin D and calcium, and the pain and serum calcium and phosphate values have improved to the point that he can receive out-patient treatment. PMID- 10830181 TI - Membranoproliferative glomerulonephritis and demyelinating neuropathy caused by type II mixed cryoglobulinemia associated with HCV infection. AB - Type II cryoglobulinemia is sometimes observed in patients with HCV infection. It is also well known that some cases with membranoproliferative glomerulonephritis (MPGN) possess cryoglobulinemia, which can induce neuropathy. We treated a 73 year-old woman with hepatitis C virus (HCV) who was admitted to our department because of proteinuria. She was diagnosed as MPGN according to renal biopsy. Her ankle jerk was absent with reduced touch sensation and vibration below the knees; sural nerve biopsy revealed demyelinating neuropathy. Administration of prednisolone resulted in improved proteinuria. Cryoglobulinemia associated with HCV infection seemed to induce MPGN and demyelinating neuropathy. PMID- 10830182 TI - Acute eosinophilic pneumonia associated with smoke from fireworks. AB - We report a case of acute eosinophilic pneumonia (AEP). Although the patient had been a habitual cigarette smoker for over 4 months, he had had not any respiratory distress. After he inhaled smoke from fireworks for 3 consecutive nights, the patient began to complain of cough, fever and dyspnea. He showed leukocytosis of 16,200/microl and hypoxemia of 58.1 torr. Chest radiograph showed bilateral infiltrates with Kerley A and B lines. The bronchoalveolar lavage fluid revealed 38.5% eosinophils. He was diagnosed as AEP. In this patient, inhaling of smoke from fireworks was clinically suspected to be associated with the induction of AEP. PMID- 10830183 TI - An autopsy case of diffuse panbronchiolitis associated with lung cancer. AB - The prognosis of diffuse panbronchiolitis (DPB) has been remarkably improved after the development of low-dose erythromycin therapy, possibly due to anti inflammatory rather than anti-infective mechanisms. Interestingly, DPB associated with lung cancer is quite rare. Here, we report an autopsy case of DPB who developed lung cancer after a long successful therapy with low-dose erythromycin. PMID- 10830184 TI - Clinical features of three fatal cases of non-specific interstitial pneumonia. AB - We describe the clinical courses of the 3 fatal patients (2 females and 1 male) with idiopathic non-specific interstitial pneumonia (NSIP) among 24 patients with NSIP. Lung biopsies were diagnosed to be NSIP group II in all patients. The clinical courses from onset to death of these 3 patients were 41 months, 46 months, and 91 months. A follow-up chest CT demonstrated no apparent honey-comb formation. We found that i) about 20% of patients with NSIP died of respiratory failure, ii) in the chest CT findings, apparent honey-comb formation was rare even just before death, iii) prediction of the prognosis based on the histological findings was difficult. This is the first report to describe the clinical features of deceased patients with idiopathic NSIP; the incidence of fatal cases was considered to range from 10 to 20%. PMID- 10830185 TI - Fusion of the nucleoporin gene, NUP98, and the putative RNA helicase gene, DDX10, by inversion 11 (p15q22) chromosome translocation in a patient with etoposide related myelodysplastic syndrome. AB - We report a 50-year-old man who developed therapy-related myelodysplastic syndrome after treatment with etoposide-including chemotherapy for extratesticular germ cell tumor. Chromosomal analysis showed inversion 11 (p15q22) translocation. Reverse transcriptase-polymerase chain reaction amplification of patient RNA showed a fusion transcript of nucleoporin gene NUP98, and putative DEAD-box RNA helicase gene DDX10. NUP98 is implicated in the transformation through aberrant nucleocytoplasmic transport. DDX10 is suggested to be involved in ribosome assembly. The NUP98-DDX10 fusion transcript may promote the development of secondary hematological malignancies caused by DNA topoisomerase II inhibitors through aberrant nucleocytoplasmic transport and/or alteration in ribosome assembly. PMID- 10830186 TI - Tumor-like splenic extramedullary hematopoiesis in a patient with myelofibrosis. AB - A 61-year-old woman, who was diagnosed in 1982 as having polycythemia vera, was admitted to our hospital in July 1998 because of a splenic tumor in an enlarged spleen due to myelofibrosis. As it was difficult to identify the etiology of the splenic tumor, partial splenectomy was carried out. The resected tumor proved to be an extremely proliferative lesion as the result of extramedullary hematopoiesis. Since it is difficult to diagnose the etiology of splenic tumor, the collection and analysis of reports of relevant cases may well facilitate diagnosis. PMID- 10830187 TI - Portal systemic encephalopathy presenting with dressing and constructional apraxia. AB - We report a case with portal systemic encephalopathy who presented with dressing and constructional apraxia and subtle weakness of the left hand. We initially suspected a cerebrovascular attack in the right cerebral hemisphere, but brain T1 weighted magnetic resonance (MR) imaging revealed high intensity in the basal ganglia and hyperammonemia was detected. We performed abdominal MR angiography, which visualized an intrahepatic portal systemic shunt. Cerebral blood flow, measured by xenon-enhanced computed tomography, was decreased in the bilateral, but more dominantly right-sided, parietal watershed regions. We speculate that these boundary territories might be susceptible to damage by toxic metabolites of hepatic encephalopathy. PMID- 10830188 TI - Hypercoagulopathy with piperacillin administration in osteomyelitis. AB - A 51-year-old man with osteomyelitis developed acute renal failure and superior mesenteric venous (SMV) thrombosis after piperacillin (PIPC) treatment. Coagulation profile disclosed disseminated intravascular coagulation (DIC). The serum levels of IgE and eosinophil cationic protein showed significant increases, while a lymphocyte stimulation test with PIPC also demonstrated an extremely high index. These observations suggest that hypersensitivity to PIPC might play a role in the pathogenesis of acute renal failure and SMV thrombosis due to hypercoagulopathy. Withdrawal of PIPC and anticoagulation therapy resulted in clinical improvement and normalization of the affected laboratory data. This is the first report to describe PIPC-induced hypercoagulopathy. PMID- 10830189 TI - Successful combined therapy of cyclophosphamide and cyclosporine for acute exacerbated interstitial pneumonia associated with dermatomyositis. AB - Interstitial pneumonia of polymyositis and dermatomyositis (PM/DM), especially the rapid progressive type, is a serious complication. We report a case of acute exacerbated interstitial pneumonia associated with DM. The respiratory distress condition was refractory to steroid pulse therapy and cyclosporine, however, there was a good response to additional intravenous cyclophosphamide pulse therapy (IVCY). We propose the possibility that the combination of these immunosuppressants might be useful for interstitial pneumonia with DM which is resistant to conventional steroid therapies. PMID- 10830190 TI - High incidence of acute myositis with type A influenza virus infection in the elderly. AB - We describe six elderly cases of acute myositis with type A influenza virus infection (Sydney) during the 1998-1999 outbreak. All six cases suffered from myalgia or muscle weakness especially in the lower extremities and the serum creatine kinase (CK) values were elevated above 1,000 IU/l without MB isoenzymes or electrocardiogram abnormalities. There have been a few case reports of acute myositis associated with influenza virus infection in the elderly. However, we noticed a high incidence of acute myositis among elderly patients with type A influenza virus infection. This complication may occur more commonly in elderly patients than has previously been thought. PMID- 10830191 TI - Paragonimiasis westermani with multifocal lesions in lungs and skin. AB - We report a case of Paragonimus westermani infection with a reticulonodular lesion in the right lung, left pleural effusion, and a mobile subcutaneous mass. Analyses of pleural effusion and bronchoalveolar lavage fluid (BALF) showed marked eosinophilia and high levels of eosinophil cationic protein and interleukin (IL)-5. Transbronchial lung biopsy revealed the presence of pneumonia with mild eosinophilic infiltration but remarkable lymphocytic infiltration. In this patient, high IL-5 levels in both BALF and pleural effusion could explain the remarkable eosinophilia. PMID- 10830192 TI - Diffusion and perfusion magnetic resonance imaging in childhood stroke. AB - Two magnetic resonance imaging techniques, diffusion and perfusion imaging, are being used increasingly for evaluation of pathophysiology of stroke. This article introduces these techniques and reports some initial studies using these approaches, together with conventional T2-weighted magnetic resonance imaging, for investigation of childhood stroke. It is shown that the combination of T2 weighted and diffusion images can provide information about the timing of stroke events in childhood, and perfusion imaging can detect abnormalities not visible by other magnetic resonance imaging techniques. These magnetic resonance methods therefore should play an important role in investigation of children with stroke and could be of particular value in studies of at-risk populations of children such as those with sickle cell disease. PMID- 10830193 TI - Nontraumatic brain hemorrhage in children: etiology and presentation. AB - The clinical and radiographic findings of 68 children and adolescents with nontraumatic intraparenchymal brain hemorrhage were analyzed retrospectively. There were 43 boys and 25 girls, and the average age was 7.1 years (range, 3 months to 18 years). The most common presenting symptom was a combination of headache or vomiting (40 cases, or 58.8%). Hemiparesis was the major presenting sign in 11 (16.2%) of the children, seizures occurred in 25 (36.8%) patients, and 6 (8.8%) children were irritable. Only 2 (2.9%) children were comatose at presentation. One or more risk factors for hemorrhage were found in 61 (89.7%) of 68 children. A third (23 cases, or 33.8%) had an arteriovenous malformation or fistula; altogether 29 (42.6%) children had some type of congenital vascular anomaly. Hematologic or coagulation disorders were present in 22 (32.4%) patients, and 9 (13.2%) patients had brain tumors. Hemorrhage could not be attributed to systemic hypertension in any child. The likelihood of establishing the cause of bleeding was greater when evaluation included cerebral angiography (97.3% versus 80.4% without angiography). Half (34 cases, or 50.0%) of the patients regained normal neurologic function. Six (8.8%) patients died, either directly or partly as a consequence of the hemorrhage. The remaining patients had various neurologic sequelae, including 17 (25.0%) with hemiparesis, 5 (7.4%) with aphasia, 7 (10.3%) with epileptic seizures, and 3 (4.4%) with hydrocephalus. More detailed follow-up studies are needed to obtain more information about the frequency of cognitive sequelae. PMID- 10830194 TI - Stroke in childhood: outcome and recurrence risk by mechanism in 59 patients. AB - This paper describes 59 patients, 3 months to 16 years of age, who were seen consecutively in the same center for cerebral arterial infarction. It focuses on the mechanism of stroke. The pathophysiologic process could be established for 78% of the children. Arteriopathic stroke (31 patients, or 53%) was the most common. The arteriopathies were either progressive (moyamoya in 4 patients, or 7%) or nonprogressive (27 patients, or 46%). The latter form occurred in two patterns: dissection of cervicocephalic arteries (12 patients, or 20%) and transient cerebral arteriopathy of unknown origin but probably angiitis (15 patients, or 25%). Cardiac or transcardiac embolic stroke occurred in 12% of the series and systemic diseases in 14%. There was a favorable outcome in 70% of patients having stroke due to nonprogressive arterial disease and stroke due to unidentified mechanisms. In contrast, only 26% of patients with embolic stroke, systemic disease, or moyamoya had a favorable outcome. Recurrences were more frequent and severe in this latter group. It is concluded that it is important to determine the mechanism of childhood stroke, because it strongly influences outcome, the recurrence risk, and treatment choice. PMID- 10830195 TI - Children with stroke: polymorphism of the MTHFR gene, mild hyperhomocysteinemia, and vitamin status. AB - The aim of this study was to investigate a possible association among the thermolabile polymorphism, nucleotide 677 cytosine to thymidine point mutation (677 C-->T) of the methylenetetrahydrofolate reductase (MTHFR) gene, hyperhomocysteinemia, serum folate, vitamins B12 and B6, and stroke in children. Allele and genotype frequencies for the 677 C-->T polymorphism in 21 children with stroke and 28 healthy children of the same age were studied. No differences in allelic frequency were detected between the two populations. However, the prevalence of homozygous 677 C-->T was doubled in the stroke population (28.6%) compared to the healthy group (14.3%). Total plasma homocysteine (tHcy) levels were significantly increased in children aged 2 months to 15 years with stroke compared to reference values. No association was observed between the homozygous genotype (T/T) and hyperhomocysteinemia, nor between the T/T genotype and low folate levels (below the 95th percentile) in this group of patients. Vitamin concentrations in patients were not significantly different from reference values. Significant negative correlations were found between tHcy and folate and between tHcy and cobalamin, but not between tHcy and B6 concentrations. In summary, a higher prevalence of hyperhomocysteinemia and the 677 C-->T polymorphism were observed in children with stroke, but were not always associated. The systematic study of both abnormalities in children with stroke is recommended, so that hyperhomocysteinemia of any genetic origin can be corrected with vitamin supplementation. Moreover, the 677 C-->T genotype is a strong factor for predisposition to hyperhomocysteinemia and recurrent risk of stroke that might also be prevented with folate supplementation. PMID- 10830196 TI - Risk factors for arterial ischemic stroke in children. AB - Since early recurrence occurs in at least 10% of patients presenting with their first stroke in childhood in the reported series, the search for modifiable risk factors should be a priority. Risk factors for stroke in adults include hypertension, diabetes, and smoking, as well as cardiac disease and sickle cell anemia; asymptomatic cerebrovascular disease and transient ischemic events may predict stroke in this age group. The investigation of a child with a stroke has traditionally focused on finding a single cause rather than looking for risk factors to which the patient may be exposed life long. Approximately half of children presenting with stroke have a known predisposing condition, but some have unexpected pathologies such as primary cerebrovascular disease associated with congenital heart anomalies, or may have modifiable risk factors such as hypertension associated with sickle cell disease. The literature on children presenting with initially unexplained (cryptogenic) stroke suggests that there is a daunting list of possible causes, but since the series have mainly been small, it has been difficult to evaluate the relative importance of the reported associations. This paper reviews the literature on congenital, genetic, and acquired risk factors for stroke in childhood, and includes data from the large series of patients seen at Great Ormond Street Hospital over the past 10 years. The majority have arteriographic abnormalities and there is little evidence for asymptomatic cardiac disease. Genetic predisposition, trauma, infection, and nutritional deficiencies appear to be important, although case-control studies will be required to prove causation. Appropriate screening for modifiable risk factors may lead to prevention of recurrence in some patients. In the long term, an understanding of the multiple etiologies of childhood cerebrovascular disease and ischemic stroke may lead to primary prevention in this age group, and perhaps in adults. PMID- 10830197 TI - Stroke in children: genetic and metabolic issues. AB - The study of genetic and metabolic etiologies of pediatric stroke, both vascular and metabolic, allows an understanding of the causes of acute focal neurologic deficits in childhood. Here, the mendelian and mitochondrial genetic causes of pediatric stroke syndromes are reviewed. This approach elucidates the etiology of childhood stroke and illustrates many of the genetic risk factors that are found in adult-onset cerebrovascular disease. Therefore, the study of childhood stroke serves as a model to elucidate the potential risk factors for all stroke. Ultimately this will serve to develop a more rational preventive and therapeutic approach for all cerebrovascular disease. PMID- 10830198 TI - Neurologic outcome in survivors of childhood arterial ischemic stroke and sinovenous thrombosis. AB - Ischemic stroke during infancy and childhood has the potential for life-long morbidity. Information on the neurologic outcome of children who survive ischemic stroke is lacking. Children surviving ischemic stroke between January 1, 1995 and July 1, 1999 were prospectively followed. Neurologic deficit severity was based on the Pediatric Stroke Outcome Measure (PSOM) developed in this study and parental response to two recovery questions. Predictor variables for poor outcome were tested. One-hundred twenty-three children with arterial ischemic stroke and 38 with sinovenous thrombosis were followed for a mean of 2.1 years (range, 0.8 to 6.6 years). The primary outcome based on PSOM assessment was: normal, 37%; mild deficit, 20%; moderate deficit, 26%; and severe deficit, 16%. The secondary outcome was full recovery in 45% of patients, based on parental response. The primary and secondary outcome measures were moderately correlated (P < .001; K = 0.5). In bivariate analysis, arterial stroke type, male gender, age of at least 28 days, presence of associated neurologic disorders, and need for rehabilitation therapy after stroke were predictors of poor outcome (P < .05). Multivariate analysis showed that only arterial ischemic stroke, associated neurologic disorders, and presence of rehabilitation therapy were independent predictors of poor outcome (P < .02). Poor outcome in children after ischemic stroke is therefore frequent and more likely in the presence of arterial stroke, rehabilitation therapy, and associated neurologic disorders, which justifies clinical trials of treatment strategies in childhood ischemic stroke. PMID- 10830199 TI - Intelligence after stroke in childhood: review of the literature and suggestions for future research. AB - Review of published clinical and neuropsychologic outcome studies reveals limited information about intellectual functioning after childhood stroke. The extant data are supplemented here by analysis of intelligence quotient (IQ) results obtained from 38 children in an ongoing study of unilateral middle cerebral artery ischemic stroke. Evidence so far indicates that, after stroke, mean IQ falls significantly below the population mean but remains within the average range. There is no significant difference between hemispheric side of injury; the Verbal and Performance IQ lateralization profile widely recognized in adults with unilateral injury is not apparent in younger children, and there is only a trend toward this profile in older children. The effects of a number of other variables, including sex, site of stroke, and longitudinal assessment, are also considered. Although the generally minor effect of stroke on IQ is encouraging, a number of children do require extra help on return to school. Some suggestions for future research are highlighted in order to encourage further consideration of the issues raised here. PMID- 10830200 TI - Multicenter prospective study of children with sickle cell disease: radiographic and psychometric correlation. AB - After obtaining familial informed consent, between January 1996 and July 1997, 173 children (5 to 15 years old) with sickle cell disease were enrolled in a prospective multicenter study using blood screening, transcranial Doppler ultrasonography (n = 143), cerebral magnetic resonance imaging (n = 144), and neuropsychologic performance evaluation (n = 156) (Wechsler Intelligence tests WISC-III, WIPPSI-R), which were also performed in 76 sibling controls (5 to 15 years old). Among the 173 patients with sickle cell disease (155 homozygous for hemoglobin SS, 8 sickle cell beta0 thalassemia, 3 sickle cell beta+ thalassemia, 7 sickle cell hemoglobin C disease SC), 12 (6.9%) had a history of overt stroke, and the incidence of abnormal transcranial Doppler ultrasonography (defined as mean middle cerebral artery velocity > 200 cm/sec or absent) was 8.4% in the overall study population and 9.6% in patients with homozygous sickle cell anemia The silent stroke rate was 15%. Significantly impaired cognitive functioning was observed in sickle cell disease patients with a history of stroke (Performance IQ and Full Scale IQ), but also in patients with silent strokes (Similarities, Vocabulary, and Verbal Comprehension). However, infarcts on magnetic resonance imaging were not the only factors of cognitive deficit: Verbal IQ, Performance IQ, and Full Scale IQ were strongly impaired in patients with severe chronic anemia (hematocrit < or = 20%) and in those with thrombocytosis (platelets > 500 x 10(9)/L). Multivariate logistic regression analysis showed that abnormal magnetic resonance imaging (odds ratio [OR] = 2.76) (P = .047), hematocrit < or =20% (OR = 5.85) (P = .005), and platelets > 500 x 10(9)/L (OR = 3.99) (P = .004) were independent factors of cognitive deficiency (Full Scale IQ < 75) in sickle cell disease patients. The unfavorable effect of low hematocrit has already been suggested, but this is the first report concerning an effect of thrombocytosis and showing that silent stroke alone is not a factor of cognitive deficit when not associated with low hematocrit or thrombocytosis. The effect of hydroxyurea, which is known to increase hematocrit and decrease platelet count, on cognitive functioning of sickle cell patients should be evaluated prospectively. PMID- 10830201 TI - Lessons from the Stroke Prevention Trial in Sickle Cell Anemia (STOP) study. AB - Stroke is generally uncommon in children, but sickle cell disease is associated with a high risk of stroke in the early years of childhood. Large cerebral arteries, especially the middle cerebral and intracranial internal carotid, develop stenosis that predisposes to ischemic stroke. Noninvasive prediction of risk using transcranial Doppler ultrasonography made it possible to test primary stroke prevention in a clinical trial comparing chronic blood transfusion with standard care. A consortium of 14 clinical centers conducted a randomized clinical trial (Stroke Prevention in Sickle Cell Anemia--the "STOP" study) to test a strategy to prevent first stroke in children with sickle cell disease. Over 2000 children were screened with transcranial Doppler ultrasonography and of these, 130 with elevated blood velocity indicating high risk were enrolled in the trial. Regular red cell transfusions sufficient to reduce the percentage of Hb S gene product from over 90 to less than 30 of total hemoglobin was associated with a marked reduction in stroke. The untreated risk of 10% per year was reduced over 90% with treatment, an effect sufficient to cause early termination of the trial. Although treatment was unblinded, the design included blinded adjudication of possible stroke by a panel of neurologists remote from the study sites. The study led to a Clinical Alert, issued by the National Heart, Lung, and Blood Institute, recommending screening and consideration of treatment in children with sickle cell disease and 2 to 16 years of age who are at risk based on transcranial Doppler ultrasonography, and who have not had stroke. PMID- 10830202 TI - Rational approach to treatment of moyamoya disease in childhood. AB - Early diagnosis and treatment of moyamoya disease in children is essential to minimize residual mental and physiologic deficits. Current treatment of childhood moyamoya disease in Japan, preoperative evaluation of perfusion reserve as a surgical indication, and the role of noninvasive follow-up by magnetic resonance angiography are reported. Approximately 20% of children with definite moyamoya disease were observed or treated medically. Among surgical procedures, single indirect bypass surgery was used in approximately 30% of all patients; combinations of direct and indirect bypass surgery, 20%; and multiple-indirect bypass surgery, 18%. Both adequate understanding of the primary condition and determination of optimal treatment, including specific operative procedures, required evaluation of cerebral circulation and metabolism. Surgical indications included reduced perfusion reserve in affected brain by positron emission tomography or single photon emission tomography with administration of acetazolamide or a CO2 load. Postoperative improvements of cerebral perfusion reserve show better correlation with disappearance of ischemic attacks than does angiographically demonstrated collateral formation. Follow-up evaluation with magnetic resonance angiography has advantages over conventional angiography because it is noninvasive and avoids general anesthesia. PMID- 10830203 TI - The role of transforming growth factor beta in chronic renal allograft nephropathy. PMID- 10830204 TI - Statins for all. PMID- 10830205 TI - HHV-8: a newly recognized pathogen in transplantation. PMID- 10830206 TI - Preflush with plasminogen activator in non-heart-beating donors: is it worth it? PMID- 10830207 TI - Renal transplant protocol biopsies: a surrogate biomarker for late graft loss. PMID- 10830208 TI - Changing perspectives in portal vein thrombosis and liver transplantation. PMID- 10830209 TI - The challenge of graft-specific immunosuppression in xenotransplantation. PMID- 10830210 TI - Outcome of kidney transplant recipients with previous human herpesvirus-8 infection. AB - BACKGROUND: The consequences of a prior human herpesvirus-8 (HHV-8) infection in kidney-transplant recipients are still partially unknown. The aim of this monocentric study was to determine the prevalence of HHV-8-seropositive patients at the time of transplantation and to identify the main clinical events of these HHV-8+ recipients. METHODS: From January 1, 1990 to December 31, 1996, antibodies to HHV-8 latent nuclear antigen were detected by indirect immunofluorescent method in serum samples collected just before kidney transplantation from 400 consecutive patients. Conventional double or triple immunosuppressive treatment was prescribed. For the group of HHV-8+ recipients, data including death rate, graft survival, and occurrence of Kaposi's sarcoma (KS) were retrospectively collected until December 31, 1998. Cofactors associated with KS were studied in univariate and multivariate analyses using a Cox model. RESULTS: Thirty-two patients (8%) had antibodies to HHV-8 in their sera at the time of transplantation. Among these 32, 3 years after transplantation, graft survival was 72%, and KS prevalence was 28% (KS incidence: 8.2/yr/100 HHV-8+ recipients). Multivariate analysis identified bacterial and/or Pneumocystis carinii infection (odds ratio: 8.6; P=0.019) and female gender (odds ratio: 5.34; P=0.047) as factors associated with KS. No KS was observed in patients without anti-HHV-8 antibodies at the time of transplantation. CONCLUSIONS: The low graft survival and the high prevalence of KS within the studied population of HHV-8+ transplant recipients are strong arguments for systematic screening of HHV-8 serologic features before transplantation, especially in patients of African origin. HHV-8+ transplant recipients should be closely monitored to severe infections. PMID- 10830211 TI - Warm preflush with streptokinase improves microvascular procurement and tissue integrity in liver graft retrieval from non-heart-beating donors. AB - BACKGROUND: Apart from the warm ischemic insult, integrity of liver grafts from non-heart-beating donors (NHBD) is additionally affected by microvascular alterations including erythrocyte aggregation and thrombus formation, which might hamper appropriate equilibration of the preservation solution to the grafts' microvasculature precluding cold preservation. Thus, the objective of our study was to characterize microvascular perfusion quality of University of Wisconsin (UW) solution during initial flushout of livers from NHBD rats, and to analyze the effects of an additional warm preflush with Ringer's lactated solution (RL) and with RL containing streptokinase (SK). METHODS: Cardiocirculatory arrest was induced by phrenotomy. Subsequent to 30 min of warm ischemia, livers were perfused via an aortic catheter by gravity of 100 cm H2O either with 4 degrees C 100 ml UW solution (UW, n=7), or with 25 degrees C 30 ml RL preflush followed by 4 degrees C 100 ml UW solution (RL+UW, n=7), or with 25 degrees C 30 ml SK- (7500 IU) containing RL preflush and 4 degrees C 100 ml UW solution (SK/RL+UW, n=6). Liver microperfusion was quantified using in situ fluorescence epi-illumination microscopy. Liver microcirculation of sham-operated living animals (n=4) served as controls. Enzyme release after a 24-hr cold preservation period was used as an indicator of graft integrity. RESULTS: After 30 min of warm ischemia, microvascular perfusion of UW solution was found markedly altered when compared with that of sham-operated living controls, as indicated by a significant reduction (P<0.05) of acinar and sinusoidal perfusion. Preflush with RL (RL+UW) only slightly attenuated the acinar and sinusoidal perfusion deficits, whereas the addition of SK to RL (SK/RL+UW) resulted in a significant improvement of microvascular graft perfusion (P<0.05). Accordingly, the increased enzyme release observed in solely UW-flushed livers after 24 hr cold preseravtion was only slightly influenced by preflush with RL, but markedly attenuated (P<0.05) by pre flush with RL containing SK. CONCLUSION: The additive fibrinolytic therapy using SK is effective to improve microvascular procurement of livers after warm ischemia and may thus represent a promising approach to attenuate parenchymal cell injury in liver graft retrieval from NHBD. PMID- 10830212 TI - Lung deflation impairs alveolar epithelial fluid transport in ischemic rabbit and rat lungs. AB - BACKGROUND: Because the fluid transport capacity of the alveolar epithelium after lung ischemia with and without lung deflation has not been well studied, we carried out experimental studies to determine the effect of lung deflation on alveolar fluid clearance. METHODS: After 1 or 2 hr of ischemia, we measured alveolar fluid clearance using 125I-albumin and Evans blue-labeled albumin concentrations in in vivo rabbit lungs in the presence of pulmonary blood flow and in ex vivo rat lungs in the absence of any pulmonary perfusion, respectively. RESULTS: The principal results were: (1) lung deflation decreased alveolar fluid clearance while inflation of the lungs during ischemia preserved alveolar fluid clearance in both in vivo and ex vivo studies; (2) alveolar fluid clearance was normal in the rat lungs inflated with nitrogen (thus, alveolar gas composition did not affect alveolar fluid clearance); (3) amiloride-dependent alveolar fluid clearance was preserved when the lungs were inflated during ischemia; (4) terbutaline-simulated alveolar fluid clearance was preserved in the hypoxic rat lungs inflated with nitrogen; (5) lecithinized superoxide dismutase, a scavenger of superoxide anion, and N(omega)-nitro-L-arginine methyl ester, an inhibitor of nitric oxide, preserved normal alveolar fluid clearance in the deflated rat lungs. CONCLUSION: Lung deflation decreases alveolar fluid clearance by superoxide anion- and nitric oxide-dependent mechanisms. PMID- 10830213 TI - Reperfusion lung injury after cold preservation correlates with decreased levels of intrapulmonary high-energy phosphates. AB - BACKGROUND: To evaluate the role of energy state in primary graft dysfunction, which is crucial in lung transplantation, we investigated the relationship between intrapulmonary high-energy phosphate compounds and reperfusion lung injury after cold preservation. METHODS: Using an isolated rat lung perfusion model with fresh rat blood as perfusate, rat lungs were exposed to various cold preservation periods (0, 6, 9, and 12 hr) and reperfused. RESULTS: We found that extending the preservation period exacerbated the pulmonary hemodynamics after reperfusion. The levels of intrapulmonary high-energy phosphate compounds did not change during cold preservation, but these levels after reperfusion decreased as the preservation period was prolonged. The pulmonary hemodynamics after reperfusion were inversely correlated with the intrapulmonary high-energy phosphate compound levels after reperfusion. Total adenine nucleotide and ATP were sensitive indicators of reperfusion lung injury after cold preservation. Energy charge was not a sensitive indicator. The decreased levels of intrapulmonary high-energy phosphate compounds after reperfusion following cold preservation period were partially caused by their decreased production. CONCLUSIONS: These results demonstrated that reperfusion lung injury after cold preservation was closely correlated with decreased levels of intrapulmonary high energy phosphate compounds after reperfusion, although the levels of the intrapulmonary high-energy phosphate compounds did not change during cold preservation of up to 12 hr. PMID- 10830214 TI - Toxicity of cationic liposome-DNA complex in lung isografts. AB - BACKGROUND: Cationic lipids have been successfully employed as vectors for gene transfer in lung grafts, yet those lipid vectors have potential toxicity. Furthermore, the optimal concentration of cationic lipids for gene transfection to lung grafts has not been determined. We evaluated liposome concentration/toxicity relationships in an in vivo rat lung transplantation model. METHODS: Left lungs were harvested and infused via the pulmonary artery with chloramphenicol acetyl-transferase (CAT)-DNA/lipid 67 (cationic lipid)/dioleoylphosphatidylethanolamine complex (4:1:2 in a final concentration ratio). Donor lungs were allocated into six groups according to lipid 67 concentration: group 1, 0 microM (control); group 2, 10 microM; group 3, 50 microM; group 4, 100 microM; group 5, 250 microM; group 6, 500 microM. Forty eight hours after orthotopic transplantation, the recipient contralateral right main pulmonary artery and bronchus were ligated. The graft was ventilated with 100% oxygen for 5 min. Arterial blood gas analysis (PaO2, PaCO2), peak airway pressure (PAP), and CAT activity of the grafts were measured. RESULTS: Recipient survival, and PaO2, PAP, and CAT levels correlated with the lipid-DNA complex concentration. The grafts in groups 4-6 were more injured as evidenced by decreased PaO2 and increased PAP levels in comparison to the control group. CAT level was significantly lower in group 2 than in groups 3-6. CONCLUSIONS: The pulmonary toxicity of cationic lipid is dose-dependent. The balance between lung graft function and transgene expression is optimal at a lipid 67 concentration of 50 microM. PMID- 10830215 TI - Protective effect of CTLA4Ig secreted by transgenic fetal pancreas allografts. AB - BACKGROUND: Pancreas allotransplantation offers a cure for insulin-dependent diabetes mellitus. Systemic immunosuppression used to prevent immune destruction of the graft has side-effects, including increased susceptibility to infection and neoplasia. These unwanted effects may be limited by engineering the graft to secrete immunomodulatory molecules, to achieve local immunosuppression. Several studies have shown that transient local CTLA4Ig results in partial protection of allogeneic grafts. Our intent has been to determine whether sustained secretion of transgenic CTLA4Ig from pancreatic islets is able to protect against allograft rejection. METHODS AND RESULTS: Mouse CTLA4 (test=CTLA4Ig) or CD5 leader sequence (control=CD5LIg) was fused to the Fc of mouse IgG2c, and expressed transgenically under the control of the rat insulin promoter in C57BL/6 mice carrying the bml mutation of H-2K(b) (B6.C-H-2(bm1)). This resulted in expression in pancreatic islets. We used ELISA quantification of transgene products secreted into the supernatants of cultured fetal pancreata to select high (CTLA4Ig(hi)) and low (CTLA4Ig(lo)) expresser transgenic mice. Cultured fetal pancreata were transplanted under the kidney capsule of wholly allogeneic CBA recipient mice. CTLA4Ig(hi) but not CTLA4Ig(lo) expresser grafts showed enhanced survival compared with control CD5LIg grafts at 6 weeks posttransplant, provided the recipient mice were transiently depleted of CD4 T cells (by a single low-dose injection of GK1.5) before transplantation. CONCLUSIONS: Sustained local secretion of CTLA4Ig from transgenic grafts in combination with transient systemic CD4 T-cell depletion can enhance allograft acceptance. PMID- 10830216 TI - A non-cleavable mutant of Fas ligand does not prevent neutrophilic destruction of islet transplants. AB - BACKGROUND: Fas ligand (FasL) mediates apoptosis of susceptible Fas-expressing lymphocytes, and may contribute to the maintenance of peripheral tolerance. In transplantation models, however, artificial expression of FasL on cellular as well as islet transplants results in accelerated rejection by neutrophils. The mechanism of the neutrophilic response to FasL expression is unknown. FasL, like other members of the tumor necrosis factor family, is cleaved to a soluble form by metalloproteases. We tested the hypothesis that soluble FasL (sFasL) was responsible for neutrophil migration by creating a non-cleavable mutant of FasL. METHODS: Three mutants of FasL with serial deletions in the putative proteolytic cleavage site of human FasL were made using inverse polymerase chain reaction. The relative fractions of sFasL and membrane-bound FasL were assessed by Western blot and immunoprecipitation, as well as by cytotoxicity assay using Fas expressing target cells. The fully non-cleavable mutant was transduced into murine islets as well as myoblasts and tumor cell lines, and tested in a murine transplantation model. RESULTS: Serial deletions in the putative metalloprotease site of FasL resulted in a fully non-cleavable mutant of FasL (ncFasL). Expression of ncFasL in tumor lines induced higher levels of apoptosis in Fas bearing targets than wild-type FasL. Transplantation of ncFasL-expressing islets under the kidney capsule of allogenic mice resulted in accelerated rejection identical to that seen with wild-type Fas ligand-expressing islets. Myoblasts and tumor cell lines expressing ncFasL also induced neutrophil infiltration. CONCLUSIONS: Membrane-bound Fas ligand is fully capable of inducing a neutrophilic response to transplants, suggesting an activation by Fas ligand of neutrophil chemotactic factors. PMID- 10830217 TI - Transplantation of human peripheral blood stem cells into fetal rhesus monkeys (Macaca mulatta). AB - BACKGROUND: Methods for assessing engraftment efficiency have been explored in a primate xenogeneic model of in utero hematopoietic stem cell transplantation. METHODS: Human peripheral blood stem cells (PBSC) were obtained by leukapheresis from a human male donor after 4 days of administration of recombinant human granulocyte-colony stimulating factor (5 microg/kg/ day). PBSC were enriched for the CD34+ population with and without T-cell depletion. The resulting mononuclear cells consisted of two cell populations, one that was stem cell enriched (0.83% CD3+ cells, 95% CD34+; group 1) and one that was stem cell enriched and T-cell depleted (<0.03% CD3+ cells, 98% CD34+; group 2). Four fetal monkeys (two per group) received either two or four i.p. injections (approximately 5x10(6) cells/injection) via ultrasound guidance every other day over a 7-day period (gestational days 50, 52, 54, and 56). One fetus in each group also received i.p. recombinant human stem cell factor (25 microg/kg) and recombinant human granulocyte-colony stimulating factor (10 microg/kg) posttransplant every 10 days from gestational day 60-150. RESULTS: Four healthy newborns were delivered at term, and specimens were analyzed by polymerase chain reaction for the human Y chromosome (birth, monthly to 6 months; blood, marrow, progenitor assays). Polymerase chain reaction results were positive for all four newborns in all specimens assessed, and flow cytometric analysis for human CD45 in marrow showed engraftment ranging from 0.1-1.7%. There was no evidence of graft-versus-host disease in any of the animals. CONCLUSION: These studies show that (1) multilineage engraftment of human PBSC can be achieved in the fetal rhesus recipient, (2) the rhesus fetus appears to tolerate relatively high numbers of human CD3+ cells, and (3) healthy chimeric rhesus infants can be delivered at term after multiple in utero procedures. PMID- 10830218 TI - Reactivation of type 1 diabetes in patients receiving human fetal pancreatic tissue transplants without immunosuppression. AB - BACKGROUND: Type 1 diabetes is a cell-mediated autoimmune disease. Successful transplantation of human fetal pancreatic tissue into type 1 diabetic patients must address both autoimmunity and allograft rejection. We investigated whether humoral and cellular responses to islet antigens could be demonstrated in the peripheral blood of type 1 diabetic subjects receiving human fetal pancreatic tissue transplants. METHODS: We investigated peripheral blood mononuclear cell (PBMC) responses, using cellular immunoblotting, and autoantibody responses to islet proteins, before transplant and at 3-month intervals after transplant, of nine long-term type 1 diabetes patients (mean disease duration of 21 years) receiving human fetal pancreatic tissue subcutaneously into the abdominal wall without immunosuppression. RESULTS: Before transplant, all nine subjects were islet cell autoantibody (ICA)-negative and seven out of nine subjects were glutamic acid decarboxylase autoantibody (GADAb)-positive. After transplant, all subjects became ICA(+) and the two patients who were GADAb(-) before transplant became GADAb(+) after transplant. Maximum PBMC reactivity to separated human fetal pancreatic proteins was observed in four patients 3 months after transplant, in one patient at 6 months, in two patients at 9 months, and in one patient at 12 months after transplant. One subject, who had PBMC reactivity to multiple islet proteins before transplant, continued to respond to multiple islet proteins throughout the study. CONCLUSIONS: We conclude that the development in the peripheral blood of ICA, GADAb, and PBMC reactivity to human fetal pancreatic proteins in the trans plant recipients is most consistent with reactivation of the type 1 diabetes disease process. PMID- 10830219 TI - Simvastatin attenuates vascular hypercoagulability in cardiac transplant recipients. AB - BACKGROUND: 3-Hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors have been shown to reduce cardiac allograft failure and to lower the incidence of transplant coronary artery disease. These effects result from as yet unknown mechanisms not clearly attributable to lipid lowering. We here report that low dose simvastatin treatment inhibits excessive expression of monocyte tissue factor (TF) and reduces the persistent hypercoagulability state seen in cardiac transplant recipients. METHODS: Fifteen consecutive heart transplant recipients receiving standard oral immunosuppression were newly assigned to a 10 mg daily simvastatin therapy. Levels of TF activity in both unstimulated and lipopolysaccharide-stimulated peripheral blood mononuclear cells drawn from transplant recipients before and under simvastatin therapy were evaluated by one stage clotting assay. RESULTS: Monocyte TF activity was found to be significantly increased in cardiac transplant recipients when compared with healthy controls. Excessive monocyte procoagulant activity was reduced in cardiac transplant recipients during simvastatin treatment. This effect occurred independently of the reduction of serum low-density lipoprotein cholesterol. As demonstrated by reverse transcriptase-polymerase chain reaction, monocyte TF reduction by simvastatin, observed in 13 of the 15 transplant recipients investigated, could be ascribed to an inhibition of monocyte TF gene transcription. The reduction of monocyte TF activity during treatment with simvastatin paralleled with the normalization of elevated levels of thrombin-antithrombin complex, prothrombin fragment F1+2, and D-dimer, which are markers of thrombin and fibrin formation indicating coagulation activation after cardiac transplantation. CONCLUSION: Inhibition of monocyte TF expression and attenuation of the persistent hypercoagulable state observed in cardiac transplant recipients during treatment with simvastatin may represent an important mechanism by which HMG-CoA reductase inhibitors protect against the development of transplant coronary artery disease. PMID- 10830220 TI - Porcine endothelium supports transendothelial migration of human leukocyte subpopulations: anti-porcine vascular cell adhesion molecule antibodies as species-specific blockers of transendothelial monocyte and natural killer cell migration. AB - BACKGROUND: In cases where hyperacute rejection has been prevented, pig to primate organ transplantation results in a delayed rejection mediated by graft infiltrating leukocytes. The migration of human leukocytes across porcine endothelium is poorly characterized, but may offer targets for species-specific antirejection therapy. METHODS: Transwell tissue culture inserts with endothelial cells growing on polycarbonate filters were used to characterize the migration of peripheral blood monocuclear cells and purified leukocyte subpopulations across pig and human endothelial cells and cell lines. Endothelial cell morphology was evaluated by scanning and transmission electron microscopy, and the contribution of different adhesion receptor pairs to transendothelial migration was evaluated by antibody blocking experiments. RESULTS: There were no evident quantitative or qualitative differences in the capacity of human and porcine endothelium to support transendothelial migration of human leukocytes [T, B, and natural killer (NK) cells, monocytes, and neutrophils]. Monocytes and large granular CD3+ lymphocytes migrated most efficiently across the endothelium. Antiporcine vascular cell adhesion molecule-1 antibodies blocked transendothelial migration of human monocytes and NK cells across tumor necrosis factor-alpha stimulated pig endothelium by at least 60%. Anti-CD18 antibodies had no effect on the migration of human NK cells across pig endothelium, whereas they partly blocked migration of NK cells across human endothelium and migration of monocytes across porcine endothelium. Interleukin-2 stimulated, but not unstimulated, T and NK cells were cytotoxic to porcine endothelium. CONCLUSIONS: Porcine endothelium supports transendothelial migration of human leukocyte subpopulations as efficiently as human endothelium. Incompatibilities in some adhesion receptor pairs may be compensated for by other adhesion receptor pairs, as exemplified by human NK cells whose migration across human, but not pig, endothelium was blocked by anti CD18 antibodies. Antiporcine vascular cell adhesion molecule-1 antibodies may be used as species-specific blockers of transendothelial NK cell and monocyte migration, and as such may prove to be useful inhibitors of cellular organ xenograft rejection. PMID- 10830221 TI - Protocol renal allograft biopsies and the design of clinical trials aimed to prevent or treat chronic allograft nephropathy. AB - BACKGROUND: The minimum sample size to perform a clinical trial aimed to modify the natural history of chronic allograft nephropathy (CAN) is very large. Since the presence of chronic tubulointerstitial damage in renal protocol biopsy specimens is an independent predictor of late outcome, we evaluated whether protocol biopsies could facilitate the design of trials aimed to prevent or treat CAN. METHODS: Two hundred eighty-two protocol biopsy specimens were obtained 3 months after transplantation in 280 patients with serum creatinine levels <300 micromol/L, proteinuria <1000 mg/day, and stable function. The specimens were evaluated according to the Banff criteria. RESULTS: Graft survival depended on the presence of CAN and renal transplant vasculopathy (RTV). Thus, biopsy specimens were classified as: (a) no CAN (n=174); (b) CAN without RTV (n=87); and (c) CAN with RTV (n=21). Graft survival at 10 years was 95%, 82%, and 41%, respectively (P=0.001). Total serum cholesterol before transplantation was 4.5+/ 1.1, 4.6+/-1.1, and 5.3+/-1.6 mmol/L, respectively (P=0.009) and it was the only predictor of RTV. Power analysis (beta=20%, alpha=5%) was done to evaluate whether protocol biopsies can facilitate the design of clinical trials aimed either to prevent or treat CAN. We showed that the most feasible approach would be to use the presence of CAN as the primary efficacy end point in a prevention trial. To demonstrate a 50% reduction in the incidence of CAN at 3 months, 570 patients would be required. CONCLUSIONS: Protocol biopsies may allow a reduction of sample size and especially the time of follow-up in a trial aimed to prevent CAN. PMID- 10830222 TI - Evidence for oxidative stress in bronchiolitis obliterans syndrome after lung and heart-lung transplantation. The Munich Lung Transplant Group. AB - Bronchiolitis obliterans syndrome (BOS) is the most serious long-term sequel of lung or heart-lung transplantation (H/LTX). Neutrophilia in the lower respiratory tract is a prominent feature of BOS. Because polymorphonuclear leukocytes (PMN) are capable of releasing large quantities of reactive oxygen species, we measured indicators of oxidative stress and glutathione levels representing antioxidant defense in H/LTX patients (HLTX, n=6; double-LTX, n=7; single-LTX, n=9). We analyzed 19 bronchoalveolar lavage (BAL) samples from 13 non-BOS patients (nine female, four male: age 39+/-4 years) and 17 BAL samples from nine BOS patients (five female, four male: age 33+/-2 years). PMN were the predominant BAL cell population in BOS (61.7+/-7.8% vs. 12.3+/-3.4%, P<0.001). Myeloperoxidase activity in the epithelial lining fluid and oxidized methionine residues in BAL derived proteins were elevated in BOS (8.6+/-1.6 U/ml vs. 2.2+/-0.6 U/ml, P<0.01; and 12.6+/-1.1% vs. 7.7+/-0.8%, P<0.001, respectively). In addition, the concentration of reduced glutathione in epithelial lining fluid was decreased in BOS (162.6+/-20.1 microM vs. 345.8+/-57.1 microM, P<0.01), whereas the proportion of oxidized glutathione was increased (13.9+/-2.0O% vs. 6.7+/-1.2%, P<0.001). PMN, myeloperoxidase, and oxidized methionine residues were inversely correlated, whereas reduced glutathione was positively correlated with forced expiratory volume in 1 sec (P<0.05 to P<0.001). We conclude that excessive oxidative stress and a lack of glutathione are associated with BOS after H/LTX and may play relevant roles in the development of this disorder. PMID- 10830223 TI - A prospective randomized trial on azathioprine addition to cyclosporine versus cyclosporine monotherapy at steroid withdrawal, 6 months after renal transplantation. AB - BACKGROUND: Many attempts have been made to withdraw steroid therapy in renal transplant patients in order to avoid its many side effects. Results have been, so far, controversial. In this randomized prospective study, we compare the efficacy of azathioprine adjuncts to cyclosporine at the time of steroid withdrawal, 6 months after transplantation, versus Cyclosporine monotherapy, in preventing acute rejection. METHODS: One hundred and sixteen kidney transplant patients with good and stable renal function (creatininemia <2 mg/dl) received, in the first 6 months, cyclosporine + steroid. They were then randomized into two groups (A and B), and steroid therapy was withdrawn over 2 months. Group A (58 patients) continued on cyclosporine monotherapy, whereas group B (58 patients) added azathioprine (1 mg/kg/day) at the beginning of randomization and continued on cyclosporine + azathioprine. In both groups, patients resumed steroid therapy at the first episode of acute rejection. Follow-up after randomization was 5.3+/ 1.6 years. RESULTS: After 5 years, the incidence of steroid resumption was 57% in group A and 29% in group B (P<0.02); of those, 68% and 88% of them were within 6 months from randomization. Anti-rejection therapy was always successful. Five year patient and graft survival rates were 90% and 88% in group A and 100% and 91% in group B. Creatininemia did not differ, at follow-up. Side effects differed only for mild and reversible leukopenia caused by azathioprine in group B. CONCLUSION: Cyclosporine plus azathioprine is more effective than cyclosporine monotherapy in reducing the incidence of acute rejection after steroid withdrawal. Graft loss as a result of chronic rejection, mild in both groups, did not differ. Steroid withdrawal is feasible and advantageous, and the addition of azathioprine allowed 71% of our selected patients to remain steroid-free. PMID- 10830224 TI - The safety and efficacy of a two-dose daclizumab (zenapax) induction therapy in liver transplant recipients. AB - BACKGROUND: Induction therapy with daclizumab has been shown to be efficacious in the prevention of acute rejection in kidney transplant patients. The routine use of antibody induction therapy in liver transplantation has not gained widespread acceptance, except in the cases of renal insufficiency. The recent approval of daclizumab prompted us to initiate this pilot study using induction therapy in those patients at risk for developing posttransplant renal insufficiency. METHODS: This nonrandomized study examined the use of daclizumab in 39 of the last 97 liver transplants performed at the University of Alabama in Birmingham. The daclizumab group received 2 mg/kg intravenously before organ engraftment, and 38 of the 39 received 1 mg/kg intravenously on postoperative day 5. The control group consisted of the remaining 58 contemporary patients. Additional immunosuppression consisted of steroids, tacrolimus, or microemulsion cyclosporine in all patients and mycophenolate mofetil in selected patients. RESULTS: Pretransplant demographics were not significantly different between the groups. In the induction group there were significantly fewer males, 14 (36%) vs. 34 (59%) (P=0.03). They had greater renal insufficiency at the time of transplant, serum creatine 1.9+/-0.37 mg/dl vs. 0.8+/-0.5; P=0.0009, and more patients were at higher acuity (status 1 and 2A): 12 (31%) vs. 3 (5%) P=0.0006 than in the noninduction group. By postoperative day 7, renal function improved in the induction group such that it was not significantly different from the noninduction group and remained similar throughout the rest of the follow-up. The induction group also experienced significantly less acute rejection, 7 (18%) vs. 23 (40%) (P=0.02) than in the noninduction group in the first 6 months. The 1-, 3 , and 6-month patient survival rates were similar in the induction group, 97.4%, 97.4%, and 97.4%, vs. non-induction 94.8%, 93.0%, and 93% (P=NS). The incidence of cytomegalovirus, in the first 6 months, in the induction group was four (10%) vs. five (9%) (P=NS) in the noninduction group. CONCLUSION: In the pilot study, induction therapy with daclizumab was safe, facilitated improvement in renal function, and appeared to reduce the incidence of acute rejection. Combination therapy with daclizumab may be an important adjunct in immunosuppressive strategies for liver transplant recipients. PMID- 10830225 TI - Portal vein thrombosis in adults undergoing liver transplantation: risk factors, screening, management, and outcome. AB - BACKGROUND: Portal vein thrombosis (PVT) has been seen as an obstacle to liver transplantation (LTx). Recent data suggest that favorable results may be achieved in this group of patients but only limited information from small size series is available. The present study was conducted in an effort to review the surgical options in patients with PVT and to assess the impact of PVT on LTx outcome. Risk factors for PVT and the value of screening tools are also analyzed. METHODS: Adult LTx performed from 1987 through 1996 were reviewed. PVT was retrospectively graded according to the operative findings: grade 1: <50% PVT +/- minimal obstruction of the superior mesenteric vein (SMV); grade 2: grade 1 but >50% PVT; grade 3: complete PV and proximal SMV thrombosis; grade 4: complete PV and entire SMV thrombosis. RESULTS: Of 779 LTx, 63 had operatively confirmed PVT (8.1%): 24 had grade 1, 23 grade 2, 6 grade 3, and 10 grade 4 PVT. Being male, treatment for portal hypertension, Child-Pugh class C, and alcoholic liver disease were associated with PVT. Sensitivity of ultrasound (US) in detecting PVT increased with PVT grade and was 100% in grades 3-4. In patients with US-diagnosed PVT, an angiogram was performed and ruled out a false positive US diagnosis in 13%. In contrast with US, angiograms differentiated grade 1 from grade 2, and grade 3 from grade 4 PVT. Grade 1 and 2 PVT were managed by low dissection and/or a thrombectomy; in grade 3 the distal SMV was directly used as an inflow vessel, usually through an interposition donor iliac vein; in grade 4 a splanchnic tributary was used or a thrombectomy was attempted. Transfusion requirements in PVT patients (10 U) were higher than in non-PVT patients (5 U) (P<0.01). In hospital mortality for PVT patients was 30% versus 12.4% in controls (P<0.01). Patients with PVT had more postoperative complications, renal failure, primary nonfunction, and PV rethrombosis. The overall actuarial 5-year patient survival rate in PVT patients (65.6%) was lower than in controls (76.3%; P=0.04). Patients with grade 1 PVT, however, had a 5-year survival rate (86%) identical to that of controls, whereas patients with grades 2, 3, and 4 PVT had reduced survival rates. The 5-year patient survival rate improved from the 1st to the 2nd era in non-PVT patients (from 72% to 83%; P<0.01), in grade 1 PVT (from 53% to 100%; P<0.01), and in grades 2 to 4 PVT (from 38% to 62%; P=0.11). CONCLUSIONS: The value of US diagnosis in patients with PVT depends on the PVT grade, and false negative diagnoses occur only in incomplete forms of PVT (grades 1-2). The degree of PVT dictates the surgical strategy to be used, thrombectomy/low dissection in grade 1-2, mesoportal jump graft in grade 3, and a splanchnic tributary in grade 4. Taken altogether, PVT patients undergo more difficult surgery, have more postoperative complications, have higher in-hospital mortality rates, and have reduced 5-year survival rates. Analysis by PVT grade, however, reveals that grade 1 PVT patients do as well as controls; only grades 2 to 4 PVT patients have poorer outcomes. With increased experience, results of LTx in PVT patients have improved and, even in severe forms of PVT, a 5-year survival rate >60% can now be achieved. PMID- 10830226 TI - Donor liver uridine diphosphate (UDP)-glucuronosyltransferase-1A1 deficiency causing Gilbert's syndrome in liver transplant recipients. AB - BACKGROUND: Uridine diphosphate-glucuronosyltransferase-1A1 deficiency, causing Gilbert's syndrome, has been attributed to two extra (TA) bases in the TATAA-box of the promoter region of its gene, where the A(TA)6TAA allele corresponds to the normal gene and A(TA)7TAA corresponds to a gene with reduced expression. Our aim was to determine whether isolated hyperbilirubinemia in liver transplant recipients was due to Gilbert's syndrome acquired through the liver allograft. METHODS: From 305 patients followed in our Liver Transplant Clinic, five patients with isolated unconjugated hyperbilirubinemia in the absence of hemolysis, recurrent viral hepatitis, and biliary tract pathology were identified; 10 other post-orthotopic liver transplantion patients with normal liver chemistry tests were randomly selected as a control group. DNA was extracted from paraffin embedded liver allograft tissue and peripheral lymphocytes and was genotyped for the TA repeat at the uridine diphosphate glucononosyltransferase-lA1 promoter region by polymerase chain reaction and acrylamide gel electrophoresis. Homozygosity for the (TA)7 allele was considered diagnostic of Gilbert's syndrome. RESULTS: The mean serum total bilirubin level of the study patients was 2.28 mg/dl (range 1.8-3.0), consisting predominantly of the unconjugated form; that of the control patients was 0.76 mg/dl (range 0.4-1.1). The liver tissue from all five patients in the study group possessed the homozygous A(TA)7TAA genotype that was not observed in their lymphocytes. None of the liver tissue from the control patients demonstrated homozygosity for the A(TA)7TAA allele. CONCLUSION: Uridine diphosphate-glucuronosyltransferase-1A1 deficiency, causing Gilbert's syndrome, may be carried by the donor liver and present with isolated unconjugated hyperbilirubinemia in liver transplant recipients. PMID- 10830227 TI - Improvement of acute and chronic renal dysfunction in liver transplant patients after substitution of calcineurin inhibitors by mycophenolate mofetil. AB - BACKGROUND: Renal dysfunction caused by treatment with the calcineurin inhibitors (CNI) is a major problem in the long-term course after liver transplantation. PATIENTS: In 22 liver graft recipients with renal dysfunction and stable graft function between 3 weeks and 12 years after transplantation, CNI were substituted by MMF at a final dose of 1.5-3 g/day between October 1996 and October 1998. METHODS: In a prospective non-randomized study, the development of renal function, the side effects of MMF medication, and the stability of liver function were analyzed for a mean follow-up of 15 months. Results. (1) MMF was withdrawn in four patients for major side effects between 1 and 7 months after study entry; eight patients had minor side effects. (2) Six months after study entry, renal function had improved in 17 of the 22 study patients; mean serum creatinine +/-SD (micromol/L) was 201+/-77 at entry and 153+/-65 after 3 months (P<0.001). (3) Improvement occurred in 11 of 15 patients with creatinine elevation > or =12 months and in 6 of 6 patients with creatinine elevation < or =6 months. (4) One patient developed transient liver dysfunction and a second required retransplantation for progressive cholestasis but without signs of rejection. CONCLUSIONS: In patients who undergo liver transplantation, substitution of CNI by MMF leads to improvement of acute as well as chronic renal dysfunction in most cases. Side effects of MMF may be limiting in some patients, and the immunological consequences remain to be studied. PMID- 10830228 TI - Improved long-term results with thymoglobuline induction therapy after cardiac transplantation: a comparison of two different rabbit-antithymocyte globulines. AB - BACKGROUND: The aim of this retrospective single center analysis was to compare possible long-term benefits of two different rabbit-antithymocyte globuline (ATG) induction therapies after cardiac transplantation. PATIENTS AND METHODS: A total of 484 primary cardiac transplanted patients received induction therapy with two different rabbit-ATGs (thymoglobuline: n=342, ATG-fresenius: n=142). All patients received immunosuppressive maintenance therapy with cyclosporine, azathioprine, and prednisolone. Cardiac rejection was assessed by serial endomyocardial biopsies. Surveillance of graft arteriosclerosis was performed by angiograms 1, 3, and 5 years after transplantation. RESULTS: Five-year survival was significantly better in the thymoglobuline group (76 vs. 60%). Thymoglobuline patients had a lower rate of death from rejection (2.3 vs. 10%; P<0.01) and graft arteriosclerosis (0.88 vs. 5.6%; P<0.01). After 5 years, freedom from rejection was 72% in the thymoglobuline group compared to 42% in the ATG-fresenius group (P<0.01). Graft arteriosclerosis appeared in 14% of thymoglobuline patients and in 28% of ATG-fresenius patients (P<0.01). Viral infections occurred more often in thymoglobuline patients (53 vs. 39%, P<0.05) although there was no difference in appearance of cytomegalovirus disease (17 vs. 13%). Freedom from posttransplant malignant disease was comparable between the two groups. CONCLUSION: These results suggest that there are differences between rabbit ATG products. The superior prevention of rejection with thymoglobuline may be the reason for the lower rate of graft arteriosclerosis. PMID- 10830229 TI - Effects of antibody reactivity to major histocompatibility complex (MHC) and non MHC alloantigens on graft endothelial cells in heart allograft rejection. AB - BACKGROUND: To gain insight in the pathogenesis of vascular lesions in heart allograft rejection, we investigated effects of allosera reactive with major histocompatibility complex (MHC) or non-MHC alloantigens on graft endothelial cells (EC) in a rat transplantation model. METHODS: Anti-MHC and anti-non-MHC allosera were obtained from Brown Norway (RT.1(n)) recipients of a Lewis (RT.1(1)) or congenic LEW.1N (RT.1(n)) heart allograft respectively. Reactivity with endothelial alloantigens was studied in vitro using a series of three rat heart endothelial cell (RHEC) lines of Lewis origin. Phenotypic studies of MHC and non-MHC alloantigen expression, and adhesion molecule induction on EC were performed by immunostaining and fluorescence-activated cell sorting analysis. Complement-mediated cytotoxicity of allosera was studied using a 51Cr release assay. RESULTS: Both anti-MHC allosera and anti-non-MHC allosera showed reactivity with all three RHEC lines. EC stimulation with tumor necrosis factor alpha and interferon-y resulted in increased reactivity of anti-MHC but not of anti-non-MHC allosera. Anti-MHC allosera showed complement-mediated cytotoxicity for EC, which was strongly increased when cytokine-stimulated EC were used. With anti-non-MHC allosera, only minor cytotoxicity was measured, irrespective of the activation of EC. Anti-MHC and anti-non-MHC allosera from the day of rejection (days 7-8 and days 29-35, respectively) had similar subclass profiles of allospecific IgG, except for allospecific IgM, which was only detected in anti MHC allosera. Complement-mediated cytotoxicity of anti-MHC allosera from the day of rejection was effected mainly by IgM alloantibodies, whereas, in allosera taken 4 days after rejection, a predominance of cytotoxic alloantibodies of the IgG class was observed. No indications were found that either alloantibody reactivity alone or in combination with complement activation led to EC activation processes relevant to intercellular adhesion molecule-1 or vascular cell adhesion molecule-1 induction. CONCLUSIONS: Our data show that, in heart allograft rejection, MHC but also non-MHC alloantigens on EC are target structures in the alloantibody response. Alloantibodies reactive with endothelial MHC, but not those reactive with non-MHC alloantigens, may significantly contribute to vasculopathy by complement-mediated cytotoxicity. Although no evidence was found that alloantibodies reactive with graft EC induce adhesion molecule expression, they may trigger other EC mechanisms relevant to graft vasculopathy. PMID- 10830230 TI - Peripheral blood mononuclear cell responses from type 1 diabetic patients and subjects at-risk for type 1 diabetes to human fetal pancreatic tissue proteins. AB - BACKGROUND: Fetal pancreatic tissue has been suggested to be less immunogenic than adult islets. Thus, transplantation of human fetal pancreatic tissue as treatment for type 1 diabetes has been gaining interest. To investigate this question, we tested the peripheral blood mononuclear cell (PBMC) responses from different subject populations to human adult islet proteins (AIP) versus human fetal pancreatic proteins (FPP). METHODS: PBMC responses to FPP and AIP from normal controls (n=14), newly diagnosed type 1 diabetes patients (n=5), long-term type 1 diabetes patients (n=9), and subjects at-risk for development of type 1 diabetes (n=3) were studied. RESULTS: We observed that normal controls demonstrated PBMC reactivity to 0-3 molecular weight regions (mwr) for both the AIP (mean+/-SD, 0.8+/-1.1) and the FPP (0.6+/-0.7). In contrast, newly diagnosed type 1 diabetic patients (<1 year) demonstrated PBMC responses to 9-16 mwr for the AIP (12.8+/-2.5) and 0-14 mwr for the FPP (6.8+/-5.0). The PBMCs from long term type 1 diabetes patients (> 3 years) were responsive to 2-11 mwr for AIP (6.0+/-2.8) and 0-11 mwr for FPP (4.9+/-4.0). Three nondiabetic ICA positive subjects at-risk for development of type 1 diabetes demonstrated positive PBMC reactivity to 9-18 mwr for the AIP (12.7+/-3.9) and 4-18 mwr for the FPP (10.0+/ 5.9). CONCLUSIONS: We conclude that human fetal pancreatic proteins are not significantly less stimulatory than human adult islet proteins to PBMCs of subjects with or at risk for type 1 diabetes. PMID- 10830231 TI - Prolongation of allograft survival by Nippostrongylus brasiliensis is associated with decreased allospecific cytotoxic T lymphocyte activity and development of T cytotoxic cell type 2 cells. AB - BACKGROUND: We have demonstrated that infection with Nippostrongylus brasiliensis (Nb), which induces strong type 2 responses, prolongs kidney allograft survival in rats. Here, we confirm that this effect is not species-specific and address immune modulation in allospecific T-cell responses mediated by nematode infection. METHODS: C57BL/6 mice were injected with Nb or phosphate-buffered saline. Four days later, mice were transplanted with BALB/c hearts and graft survival was assessed. In other experiments, Nb-infected mice were immunized with BALB/c spleen cells and allospecific T-cell responses were determined in vitro. RESULTS: In this study, we show that Nb prolongs cardiac allograft survival in mice. Further, spleen T cells from Nb-infected, allo-immunized mice exhibit reduced allospecific cytotoxic T-lymphocyte activity. In contrast, allospecific proliferation of T cells in the mixed lymphocyte reaction was not reduced by Nb, ruling out immunosuppression as the mechanism of Nb-induced allograft survival. Nb infection induced IL-4 and IL-6 and inhibited IFN-gamma production by T cells in response to allo-antigen. Furthermore, anti-IL-4 treatment reduced allospecific T-cell proliferation from Nb-infected but not control mice, indicating that type 2 allospecific T cells develop in the presence of Nb. We also double-stained T cells for CD8 and IL-4 and showed that Nb induces an 8-fold increase in Tc2 cell numbers. CONCLUSIONS: These results are consistent with a hypothesis that Nb mediates prolongation of allograft survival through induction of type 2 immunity, including the development of regulatory Tc2 cells, and subsequent inhibition of allospecific cytotoxic T-lymphocyte activity. PMID- 10830232 TI - Quantification of cytotoxic T-cell gene transcripts in human lung transplantation. AB - BACKGROUND: Differentiating between acute rejection and cytomegalovirus (CMV) infection is one of the major challenges of lung transplantation. The aims of this study were to: (1) quantify the transcription of the cytotoxic T lymphocyte (CTL) effector molecules in the bronchoalveolar lavage (BAL) of lung transplant recipients and (2) evaluate the clinical usefulness of this technique. METHODS: Sixty-six single-lung, double-lung, or heart-lung transplant patients were prospectively enrolled in the study. BAL was performed either for routine surveillance or for acute graft dysfunction. RNA was extracted from BAL cell pellets and underwent competitive reverse transcription-assisted polymerase chain reaction (RT-PCR) for perforin, granzyme B, granulysin, and Fas ligand. Gene transcript analysis was compared to clinical diagnosis established by conventional methods [BAL microbiological and transbronchial biopsy (TBB) analyses]. RESULTS: After exclusion of several BAL according to the study criteria, 62 BAL were submitted for data analysis. Significantly higher expression of all the analyzed transcripts was found during CMV infection, compared with each of the other defined diagnostic categories, namely nonsignificant pathology, acute rejection, and nonviral pulmonary infection. CONCLUSION: Quantification by competitive RT-PCR of the CTL effector molecule transcripts (perforin, granzyme B, granulysin, and Fas ligand) could represent a valuable tool for the differential diagnosis of graft dysfunction in lung transplantation. PMID- 10830233 TI - Long-term follow-up of tissue-engineered intestine after anastomosis to native small bowel. AB - BACKGROUND: Our laboratory has investigated the fabrication of a tissue engineered intestine using biodegradable polymer scaffolds. Previously we reported that isolated intestinal epithelial organoid units on biodegradable polymer scaffolds formed cysts and the neointestine was successfully anastomosed to the native small bowel. The purpose of this study was to observe the development of tissue-engineered intestine after anastomosis and to demonstrate the effect of the anastomosis over a 9-month period. METHODS: Microporous biodegradable polymer tubes were created from polyglycolic acid. Intestinal epithelial organoid units were harvested from neonatal Lewis rats and seeded onto the polymers, which were implanted into the abdominal cavity of adult male Lewis rats followed by 75% small bowel resection (n=24). Three weeks after implantation, the unit/polymer constructs were anastomosed to the native jejunum in a side-to-side fashion. The anastomosed tissue-engineered intestine was measured by laparotomy 10, 24, and 36 weeks after the implantation (n= 14). During the laparotomy, all rats with an obstruction in their anastomosis were killed and excluded from the statistical analysis. Another five rats were also killed at 10 and 36 weeks for histological and morphometric studies. RESULTS: All analyzed rats survived this study and significantly increased their body weight by 36 weeks. Obstruction of the anastomosis was observed in one rat at 24 weeks and in two rats at 36 weeks; however, the anastomosis was patent in the other 11 rats by 36 weeks. The tissue-engineered intestine of these 11 rats increased in length and diameter at 10, 24, and 36 weeks after anastomosis; there were statistically significant differences between each time point except between the length of 10 and 24 weeks (P<0.016 by Wilcoxon signed rank test). Histologically the inner surface of the tissue-engineered intestine was lined with well developed neomucosa at 10 and 36 weeks; however, there were small bare areas lacking neomucosa in the tissue-engineered intestine at 36 weeks. Morphometric analysis demonstrated no significant differences in villus number, villus height, and surface length of the neomucosa at 10 and 36 weeks. CONCLUSIONS: Anastomosis between tissue-engineered intestine and native small bowel resulted in no complications after operation and maintained a high patency rate for up to 36 weeks. The tissue-engineered intestine increased in size and was lined with well developed neomucosa for the duration of the study. PMID- 10830234 TI - Analogs of 1,25-dihydroxyvitamin D3 as dose-reducing agents for classical immunosuppressants. AB - BACKGROUND: Most immunosuppressants have a narrow margin between efficacy and side effects. A major goal in the development of immunomodulatory strategies is the discovery of combinations of drugs exerting synergistic immunomodulatory effects. The active form of vitamin D, 1,25(OH)2D3, is an immunomodulator that interacts with T cells but mainly targets antigen-presenting cells. We have demonstrated synergism between 1,25(OH)2D3 and cyclosporine, rapamycin, and FK506. The aim of this study was to investigate whether this synergism could be observed with other immunosuppressants (mycophenolate mofetil, leflunomide, and the methylxanthine A802715) and whether analogs of 1,25(OH)2D3 share this synergistic capacity in vivo. METHODS: In vitro, the median effect analysis was applied to the inhibition of phytohemagglutinin A-induced lymphocyte proliferation. In vivo, synergism between analogs of 1,25(OH)2D3 and cyclosporine or mycophenolate mofetil was evaluated in experimental autoimmune encephalomyelitis. RESULTS: In vitro, all combinations with 1,25(OH)2D3 were synergistic. The strongest synergism was seen with the inhibitors of interleukin 2 secretion, cyclosporine and FK506 (indexes 0.16 and 0.27, respectively). The weakest synergism was observed in combinations using A802715, a second-signal inhibitor (index 0.52), or the nucleotide synthesis inhibitor mycophenolate mofetil (index 0.43). In vivo, analogs of 1,25(OH)2D3 share the in vitro-observed synergism with 1,25(OH)2D3. Moreover, the differences in synergism with different immunomodulators were also present in vivo, where the best synergism was again seen in combination with cyclosporine (up to 100% paralysis protection). CONCLUSIONS: These data confirm that 1,25(OH)2D3 and its analogs are potent dose reducing drugs for other immunomodulators, making them potentially interesting for clinical use in autoimmunity and transplantation. PMID- 10830235 TI - Living unrelated donors in kidney transplants: better long-term results than with non-HLA-identical living related donors? AB - BACKGROUND: Given the severe organ shortage and the documented superior results obtained with living (vs. cadaver) donor kidney transplants, we have adopted a very aggressive policy for the use of living donors. Currently, we make thorough attempts to locate a living related donor (LRD) or a living unrelated donor (LURD) before proceeding with a cadaver transplant. METHODS: We compared the results of our LURD versus LRD transplants to determine any significant difference in outcome. RESULTS: Between 1/1/84 and 6/30/98, we performed 711 adult kidney transplants with non-HLA-identical living donors. Of these, 595 procedures used LRDs and 116 used LURDs. Immunosuppression for both groups was cyclosporine-based, although LURD recipients received 5-7 days of induction therapy (antilymphocyte globulin or antithymocyte globulin), whereas LRD recipients did not. LURD recipients tended to be older, to have inferior HLA matching, and to have older donors than did the LRD recipients (all factors potentially associated with decreased graft survival). Short-term results, including initial graft function and incidence of acute rejection, were similar in the two groups. LURD recipients had a slightly higher incidence of cytomegalovirus disease (P=NS). We found no difference in patient and graft survival rates. However, the incidence of biopsy-proven chronic rejection was significantly lower among LURD recipients (16.7% for LRD recipients and 10.0% for LURD recipients at 5 years posttransplant; P=0.05). LRD recipients also had a greater incidence of late (>6 months posttransplant) acute rejection episodes than did the LURD recipients (8.6% vs. 2.6%, P=0.04). The exact reason for these findings is unknown. CONCLUSION: Although LURD recipients have poorer HLA matching and older donors, their patient and graft survival rates are equivalent to those of non-HLA-identical LRD recipients. The incidence of biopsy-proven chronic rejection is lower in LURD transplants. Given this finding and the superior results of living donor (vs. cadaver) transplants, a thorough search should be made for a living donor-LRD or LURD-before proceeding with a cadaver transplant. PMID- 10830236 TI - Acute effects of rapamycin on glomerular dynamics: a micropuncture study in the rat. AB - The acute effects of cyclosporin (CsA, 20 mg(kg i.v.) and rapamycin (RAPA, 5 mg(kg i.v.) on glomerular dynamics were separately investigated by renal micropuncture in two groups of intact rats (group CsA and RAPA, respectively) and compared with vehicle-treated rats, used as controls (group CON). Left kidney glomerular filtration rate (GFR) was decreased by CSA (-35% vs. CON, P<0.05), but was not affected by RAPA (-14% vs. CON, NS), whereas the single-nephron GFR (SNGFR) was significantly decreased in both groups (-40% in CsA, P<0.01 and -26% in RAPA, P<0.05 vs. CON). In both groups glomerular plasma flow (GPF) was significantly reduced vs. CON (CsA: -48%, and RAPA: -25%) due to the increase in both afferent (Ra) and efferent (Re) glomerular resistances: group CSA showed a prevalent rise in Re (+98% vs. CON, P<0.001) than in Ra (+66%, P<0.001); in group RAPA the increment was modest and similar in Ra and Re (+33 and +32%, respectively, NS versus CON). A further group of rats was studied in which L Arginine (ARG), the precursor of nitric oxide (NO), was administered (2.5 mg/Kg/min iv) with RAPA (group ARG). ARG limited the rise in Ra and Re, thereby preserving GPF; nevertheless, SNGFR remained low (-26% vs. CON, P<0.05) due to the decrease in the effective filtration pressure (-26% vs. CON). These data demonstrate that: (1) CsA is nephrotoxic at immunosuppressive doses; (2) RAPA, even at huge doses, has marginal effects on renal and glomerular dynamics; (3) the ARG-NO pathway is only partially involved in the vasoconstriction of superficial nephrons after RAPA administration. PMID- 10830237 TI - Beneficial effect of additional cardioplegia flush during hypothermic static cardiac preservation. AB - BACKGROUND: The effects of various preservative solutions and methods have been studied to prolong the safety period of cardiac preservation. In this study, we used cardioplegic solution (CS) during cardiac preservation and investigated how flush CS yields good preservation of isolated hearts compared with only cold immersion. METHODS: Male Wistar rat hearts were arrested with 4 degrees C St. Thomas crystalloid CS. All hearts were immersed for 6 hr in a 4 degrees C Euro Collins solution. Hearts were classified into seven groups by period and number of infusions of CS (20 ml/kg) during simple immersion of hearts. Infusion of CS during preservation was not used for group I. Infusion was performed at two hours after starting immersion for group II, at 3 hr for group III, at 4 hr for group IV, at 5 hr for group V, every hour for group VI, and every 2 hr for group VII. After preservation, the hearts were reperfused with blood using a support rat. Myocardial adenosine triphosphate was measured immediately after immersion of hearts. Biochemical examination of coronary effluents was performed at 15 min after reperfusion, and cardiac function was evaluated at 40 min after reperfusion. Myocardial specimens were subsequently taken for measurement of water content. RESULTS: Percentage recovery of left ventricular developed pressure and dp/dt in groups III, VI, and VII were higher than those in group I at each balloon volume, and left ventricular end-diastolic pressure in these groups was also significantly lower than that in group I. Levels of creatine kinase-MB and lactate in groups VI and VII after reperfusion were significantly lower than those in group I. Myocardial adenosine triphosphate was significantly better preserved in groups III, IV, VI, and VII than in group I. However, no significant difference in cardiac function or myocardial adenosine triphosphate was found among groups III, IV, VI, and VII. CONCLUSIONS: The use of CS during cardiac preservation is effective in preserving cardiac function and myocardial enzymes, and infusion may be sufficient if performed once-only at 3 or 4 hr from starting immersion in 6 hr storage of isolated rat hearts. PMID- 10830238 TI - An alternative method of arterial reconstruction after hepatic arterial thrombosis following living-related liver transplantation. AB - BACKGROUND: Hepatic artery thrombosis (HAT) remains an important cause of graft loss after liver transplantation. Emergency rearterialization methods are limited in cases of living-related liver transplantation in which the graft hepatic artery is thin and short. CASE: A 19-year-old woman who underwent living-related liver transplantation for biliary atresia developed HAT on the 4th postoperative day. During the emergency laparotomy the recipient hepatic artery was found to be too short to anastomose, so the recipient's right gastroepiploic artery was anastomosed to the graft hepatic artery. The patient is now alive and well 6 months after reoperation, and she has experienced no further episode of HAT. CONCLUSION: The right gastroepiploic artery can be used easily and safely for hepatic graft revascularization without causing ischemia of the stomach. An additional skin incision is not required, and the artery is long enough to anastomose to the graft artery directly. The method of hepatic graft rearterialization described here is an important option for patients who undergo living-related or split liver transplantation. PMID- 10830239 TI - Successful rescue of severe recurrent hepatitis C with interferon and ribavirin in a liver transplant patient. AB - BACKGROUND: Rapid graft dysfunction caused by hepatitis C virus (HCV) reinfection, although uncommon, is a disastrous complication in liver transplant patients. Finding an effective therapy for this subgroup of patients with severe recurrent HCV is a priority. METHOD: We describe a successful rescue of a 46-year old man with recurrent hepatitis C (HCV genotype 1b) using long-term interferon (IFN) and ribavirin. The patient had a very aggressive type of posttransplantation HCV infection, as judged by biochemical and histologic findings. RESULTS: Despite high pretreatment values of serum alanine aminotransferase (ALT; peak value of 901 IU/L) and HCV-RNA (2.3 x 10(6) copies/ml), the combination therapy with IFN and ribavirin produced a rapid normalization of the serum ALT values, accompanied by the clearance of serum HCV RNA. Although HCV-RNA reappeared in the serum at 3 months, the patient had continued ALT normalization and histological improvement with follow-up of over 26 months to date after the initiation of the combination therapy. CONCLUSION: This observation suggests that IFN in combination with ribavirin may offer an effective therapeutic option for liver transplant patients with severe recurrent hepatitis C. PMID- 10830240 TI - Administration of equine anti-thymocyte globulin via peripheral vein in renal transplant recipients. AB - BACKGROUND: Anti-thymocyte globulin (ATG) is used frequently as induction therapy in renal transplant recipients. It has been standard practice that the various polyclonal anti-lymphocyte preparations be administered via central line, vascular shunt, or arteriovenous fistula to avoid thrombophlebitis. METHODS: We have safely administered equine ATG (Atgam) via peripheral veins since 1978. We now retrospectively review recent experience with 83 patients transplanted between 1993 and 1998 who received Atgam induction via peripheral vein. Heparin 1000 U and Solucortef 20 mg were added to the infusion. Acetaminophen and diphenhydramine were given before infusion. RESULTS: Only two of 83 patients developed mild edema/erythema at or around the i.v. site. These reactions responded promptly to local management with warm soaks. No phlebitis or symptomatic venous thrombosis occurred in any patients. CONCLUSIONS: Equine ATG can be safely administered via peripheral vein infusion. Its use does not require continued hospitalization or placement of a central line. Peripheral vein infusion is safe, convenient, and economical. PMID- 10830241 TI - Tacrolimus and myocardial hypertrophy. AB - Tacrolimus has been used as an immunosuppressive agent in the transplantation of all solid organs. Tacrolimus-induced hypertrophic cardiomyopathy has been reported to be an unusual but serious complication. To elucidate the effects of tacrolimus on myocardial hypertrophy, we studied the relationship between the blood levels of tacrolimus and cardiac wall thickening. Our findings demonstrated that tacrolimus-induced myocardial hypertrophy correlated with tacrolimus blood levels, and that myocardial hypertrophy induced by tacrolimus was reversible. However, no patients developed clinically significant symptoms related to myocardial hypertrophy. PMID- 10830242 TI - Impact of chronic infection with chlamydia pneumoniae on incidence of cardiac allograft vasculopathy. AB - Chronic infection with Chlamydia pneumoniae (CP) is associated with development of coronary disease. However, little information exists concerning CP infection and impact on posttransplant cardiac allograft vasculopathy (CAV). A total of 202 patients were investigated 5.5+/-3.1 years after cardiac transplantation (46.5+/ 11.0 years; 169 male, 33 female). Assessment of CAV was performed by annual coronary angiograms. Chlamydia serology (IgG/IgA) was performed using micro immunofluorescence. Statistics comprised analysis of variance and Kaplan-Meier analysis. A total of 152 patients were CAV positive. Elevated titers were present in 45% (IgG) and 72.8% (IgA) of patients. Generally, serostatus was not associated with development of CAV when evaluated over the total postoperative interval. However, after month 14 there was a significant trend toward lower actuarial freedom from CAV in patients with elevated IgA titers. CP seems not to play a significant role in the development of CAV early after heart transplantation but might be a predicting risk factor after the first postoperative year. PMID- 10830243 TI - Persistence of anomalies in the growth hormone-releasing hormone-stimulated growth hormone response in diabetic-uremic patients after combined kidney pancreas transplantation. AB - Increased circulating growth hormone (GH) levels and aberrant response to different stimuli characterize both type 1 diabetes mellitus and chronic uremia and are associated with severe retinal, kidney and heart complications. Combined kidney and pancreas transplantation is a therapy that restores the endogenous, closed-loop, insulin secretion in diabetes and cure uremia. To evaluate if combined transplantation can restore a normal secretion and response of GH to growth hormone releasing hormone (GH-RH), we studied four groups of subjects: (1) seven type 1 diabetic patients with end-stage renal failure who had received pancreas and kidney transplantation (KPTx); (2) six diabetic uremic subjects, candidates for combined transplantation (IDDUP); (3) nine patients with chronic uveitis on immunosuppressive therapy comparable to pancreas recipients, six of whom treated only with prednisone (UVEST), while three (4) were treated with both prednisone and cyclosporin (UVESTCY). All subjects underwent a GH-RH test (50 microg intravenously, i.v., at 13:00 h). Serum insulin levels were significantly higher in IDDUP compared to UVEST (P=0.05) both at baseline and post GH-RH stimulus, while were similar to KPTx (P=0.2) and UVESTCY (P=0.7). In contrast, plasma free fatty acids were similar in all groups. In IDDUP baseline plasma glycerol was higher than in KPTx (P=0.04) and UVEST (P=0.02) and similar to UVESTCY (P=0.36); glycerol concentration did not change after GH-RH (P=0.08). Before and after GH-RH, serum GH levels tended to be higher in IDDUP (P=0.5) and KPTx (P=0.2) compared to UVEST and UVESTCY. Our results indicate that: 1) kidney pancreas transplantation does not normalize the GH response to GH-RH; 2) GH abnormalities are not due either to the chronic immunosuppressive therapy or to the insulin effect on GH release; 3) GH abnormalities are probably secondary to functional and/or organic complications of the hypothalamus and/or pituitary as a sequela of diabetes mellitus. PMID- 10830244 TI - CMV allograft pancreatitis: diagnosis, treatment, and histological features. AB - BACKGROUND: Cytomegalovirus (CMV) infection is a common problem in solid organ transplant recipients. CMV infection of pancreas allografts is not, however, well described. METHODS: We report the clinical presentation, histologic findings, treatment, and outcome in four patients with CMV allograft pancreatitis. These patients presented 18 weeks to 44 months after transplantation with elevated serum amylase and lipase and were suspected to have acute rejection. Percutaneous pancreas allograft biopsy specimens showed evidence of tissue invasive CMV infection. One patient had simultaneous CMV infection and acute rejection. RESULTS: Prolonged treatment with ganciclovir resulted in clinical and histologic resolution of the CMV disease. Rejection was successfully treated. Primary CMV infection in seronegative recipients seemed to be a risk factor. Three patients maintain normal allograft function; one patient lost function due to chronic rejection. The histology of tissue-invasive CMV pancreas allograft infection and its differentiation from acute rejection is described. CONCLUSION: Prompt diagnosis and prolonged therapy with antiviral agents can result in maintenance of allograft function. PMID- 10830245 TI - Fas ligand-transfected myoblasts and islet cell transplantation. AB - BACKGROUND: Expression of Fas ligand (FasL, CD95L) within the local environment of an allograft may protect from rejection by inducing apoptosis of infiltrating T cells. However, there is mounting evidence that ectopic expression of FasL stimulates an inflammatory response and targets the FasL-expressing tissue for destruction. Given the potential therapeutic applicability of FasL-based immune protection, we sought to determine whether ectopic FasL expression was detrimental and to analyze the inflammatory response induced by ectopic FasL expression in the absence of any confounding allo-immune responses. METHODS AND RESULTS: Two myoblast cell lines expressing different levels of functional FasL were produced. Co-implantation of FasL-expressing myoblasts with syngeneic islets allowed examination of the inflammatory response induced by ectopic FasL expression. In contrast to the suggested benefits of localized FasL expression, islets co-implanted with FasL-expressing myoblasts were destroyed in a vigorous inflammatory response predominated by neutrophils. Interestingly, FasL expression also had a marked anti-tumor effect. CONCLUSIONS: Unless FasL-dependent neutrophil-mediated inflammation can be prevented, it is unlikely that this strategy will be useful for preventing allograft rejection. PMID- 10830246 TI - Mycophenolate mofetil decreases endothelial prostaglandin E2 in response to allogeneic T cells or cytokines. AB - BACKGROUND: Prostaglandin E2 (PGE2) is a powerful endogenous immune suppressant and interferes with various T-cell functions. However, it is not known in detail whether immunosuppressive drugs influence the PGE2-driven immune response in transplant patients. Therefore, we investigated the effect of several immunosuppressive compounds, in particular the novel drug mycophenolate mofetil (MMF), on endothelial PGE2 release. METHODS: Endothelial cells (HUVEC) were activated by either allogeneic CD4+ or CD8+ T cells, or by the cytokines interleukin-1 or gamma-interferon. Using an enzyme-linked immunosorbent assay, we analyzed PGE2 release of the activated HWEC in the presence of MMF, cyclosporine, or tacrolimus. As verapamil and mibefradil also possess immunosuppressive properties, they were included in the study as well. RESULTS: Activation of HUVEC with interleukin-1 or T cells resulted in a drastic accumulation of PGE2 in the supernatant. Cyclosporine or tacrolimus had no effect on PGE2 release. However, Ca2+ channel blockers, when applied at higher dosages, caused a significant increase in PGE2. Interestingly, MMF strongly diminished the PGE2 level in the cell culture supernatant in a concentration-dependent manner. CONCLUSION: The results demonstrate an inhibitory effect of MMF on PGE2 production, which may lower the benefits of the PGE2-triggered immune response after organ transplantation. PMID- 10830247 TI - Intragraft proliferating T lymphocytes are associated with moderate acute pulmonary rejection. AB - Acute allograft rejection is characterized by infiltration of the donor organ by host lymphoid cells, predominantly T lymphocytes. However, the site of proliferation and clonal expansion of alloreactive T lymphocytes is not well defined in man. A group of normal transbronchial biopsies (TBB, n=9) from clinically well lung transplant recipients was compared to TBB showing acute rejection (at least grade A2, n=9), using CD3- and Ki67-specific antibodies to double-label proliferating T lymphocytes. Few double-labeled lymphocytes were present in the normal biopsies (range, 0-3 cells). However, five of the rejection biopsies contained significant numbers of proliferating T lymphocytes (range, 19 47; Fisher's exact test; P=0.029). Furthermore, this positive group contained all three cases of grade A3 rejection in the study, as well as a case with persistent grade A2 rejection on follow-up biopsy. These data demonstrate that T lymphocytes do proliferate in transplanted human lungs; such proliferation is associated with more severe rejection. PMID- 10830248 TI - The origin of alveolar macrophages in the transplanted lung: a longitudinal microsatellite-based study of donor and recipient DNA. AB - Transplanted lungs are initially populated by donor pulmonary alveolar macrophages (PAMs). These will form major antigen presenters for the recipient's suppressed immune system. They may be expected to be replaced by recipient major histocompatibility complex-compatible cells, with time. We have isolated CD14+ PAMs from bronchoalveolar lavage specimens for 6 months after transplantation and identified their origin by using microsatellite analysis. This DNA-based technology permits the reliable identification of the origin of cells from different individuals. We show that replacement of donor PAMs occurs with individual dynamics in each case. Recipient PAMs usually appeared within 2 weeks, whereas donor cells could be retained for as long as 6 months. In this limited series, there was no obvious correlation between the dynamics of this process and the occurrence of rejection episodes or infections. PMID- 10830249 TI - Recognition of islet-directed peripheral blood lymphocytes in vitro before rejection of allogeneic grafted islets. AB - A co-culture of splenic lymphocytes with allogeneic pancreatic islets [i.e., mixed lymphocyte islet co-culture (MLIC)] for 96 hr leads to reduction of beta cells and to an allospecific induction of major histocompatibility complex (MHC) class II antigens on beta-cells. The intent of our investigation was to determine whether peripheral blood lymphocytes (PBL) obtained from allogeneic islet-grafted BB/OK rats (=sensitization in vivo) cause similar alterations to donor-specific islet cells. PBL prepared before transplantation, before (at day 7) and after islet rejection were co-cultured for 24 hr with donor-specific islets. PBL obtained at any time before and after transplantation caused reduction of beta cells and enhancement of intercellular adhesion molecule-1(+)/beta-cells. Induction of MHC class II+ beta-cells was most pronounced with PBL obtained before rejection. Down-regulation of major histocompatibility complex class I+ beta-cells was caused by PBL that had been obtained from grafted animals only; it was most pronounced before islet rejection and has never been observed with lymphocytes from nongrafted normoglycemic rats. The 24-hr MLIC is capable of recognizing functionally active, donor-specific lymphocytes and is able to distinguish between the effects of sensitized and nonsensitized lymphocytes. PMID- 10830250 TI - Antibacterial use in pregnancy. PMID- 10830251 TI - Photosensitivity to ketoprofen: mechanisms and pharmacoepidemiological data. AB - The topical use of nonsteroidal anti-inflammatory drugs (NSAIDs), widely used for moderate acute and chronic painful conditions, is one of several strategies used to improve the tolerability profile of NSAIDs, particularly with regard to gastric and renal adverse effects. However, topical NSAIDs can induce photosensitivity. Among the different NSAIDs used topically, ketoprofen has often been implicated in photosensitivity reactions. Photosensitivity includes both phototoxic and photoallergic reactions. Phototoxicity can be studied in the cell system and on biological targets such as cellular membranes or DNA. In hepatocyte cultures, data suggest that radical intermediates play a role in ketoprofen photosensitised damage by cell membrane lysis. Photosensitised lysis of red blood cells has been employed as an indicator of membrane damage. Ketoprofen irradiation promotes the photolysis of erythrocyte suspensions. The drug is able to induce photoperoxidation of linoleic acid in the photo-induced lipid peroxidation process. The results obtained from the addition of radical scavengers suggest the involvement of free radicals in these processes. Ketoprofen may induce DNA damage in vitro upon irradiation. DNA, in the presence of ketoprofen, undergoes single strand breaks involving hydroxyl radicals as evidenced by the use of scavengers. Simultaneously with single strand breaks, pyrimidine dimers are formed by an energy transfer mechanism. The oxygen dependence of both processes suggest competition between a radical process leading to DNA cleavage and a poorly efficient energy transfer between ketoprofen and pyrimidines at the origin of the dimerisation process. Photoallergy is due to a cell-mediated hypersensitivity response involving immunological reactions. Therefore, it only occurs in previously sensitised individuals and requires a latency period of sensitisation. Among NSAIDs, ketoprofen is the main drug involved in this photoallergic contact dermatitis. Cross-sensitivity reactions with other arylpropionic acid derivatives, such tiaprofenic acid, fenofibrate or oxybenzone-harbouring benzoyl ketone or benzophenone may also occur. Finally, the higher frequency of such adverse reactions with ketoprofen could be accounted for by its chemical structure and the variety of chemical reactions that give rise to phototoxic effects. The widespread and repeated use of these agents may lead to sensitisation, incurring a greater risk of systemic allergic reactions with oral NSAIDs or other drugs recognised to induce cross-reactions. Physicians and pharmacists should advise patients and inform them of the risks of topical NSAIDs which are often dispensed as over the counter drugs. PMID- 10830252 TI - Risk-benefit ratio of angiotensin antagonists versus ACE inhibitors in end-stage renal disease. AB - The effective treatment of hypertension is an extremely important consideration in patients with end-stage renal disease (ESRD). Virtually any drug class--with the possible exception of diuretics--can be used to treat hypertension in the patient with ESRD. Despite there being such a wide range of treatment options, drugs which interrupt the renin-angiotensin axis are generally suggested as agents of choice in this population, even though the evidence in support of their preferential use is quite scanty. ACE inhibitors, and more recently angiotensin antagonists, are the 2 drug classes most commonly employed to alter renin angiotensin axis activity and therefore produce blood pressure control. ACE inhibitor use in patients with ESRD can sometimes prove an exacting proposition. ACE inhibitors are variably dialysed, with compounds such as catopril, enalapril, lisinopril and perindopril undergoing substantial cross-dialyser clearance during a standard dialysis session. This phenomenon makes the selection of a dose and the timing of administration for an ACE inhibitor a complex issue in patients with ESRD. Furthermore, ACE inhibitors are recognised as having a range of nonpressor effects that are pertinent to patients with ESRD. Such effects include their ability to decrease thirst drive and to decrease erythropoiesis. In addition, ACE inhibitors have a unique adverse effect profile. As is the case with their use in patients without renal failure, use of ACE inhibitors in patients with ESRD can be accompanied by cough and less frequently by angioneurotic oedema. In the ESRD population, ACE inhibitor use is also accompanied by so-called anaphylactoid dialyser reactions. Angiotensin antagonists are similar to ACE inhibitors in their mechanism of blood pressure lowering. Angiotensin antagonists are not dialysable and therefore can be distinguished from a number of the ACE inhibitors. In addition, the adverse effect profile for angiotensin antagonists is remarkably bland, with cough and angioneurotic oedema rarely, if ever, occurring. In patients with ESRD, angiotensin antagonists are also not associated with the anaphylactoid dialyser reactions which occur with ACE inhibitors. The nonpressor effects of angiotensin antagonists--such as an influence on thirst drive and erythropoiesis--have not been explored in nearly the depth, as they have been with ACE inhibitors. Although ACE inhibitors have not been compared directly to angiotensin antagonists in patients with ESRD, angiotensin antagonists possess a number of pharmacokinetic and adverse effect characteristics, which would favour their use in this population. PMID- 10830253 TI - Cardiovascular events associated with different combined oral contraceptives: a review of current data. AB - Studies of combined oral contraceptive (COC) use and cardiovascular disease have been conducted against a background of low cardiovascular risk in young women, changing COC composition and changing user selection and monitoring. Studies of myocardial infarction have found inconsistent results, possibly because of differences in the prevalence of risk factors (particularly smoking and raised blood pressure) in the populations studied. In the absence of a history of smoking and other conventional risk factors, current users of modern COCs probably do not have an increased risk of myocardial infarction. Neither are former users at risk. Evidence for important differences in the risk of myocardial infarction between formulations is weak and contradictory. Current users of low estrogen dose COCs have a small increased risk of ischaemic stroke although most of the risk occurs in women with other risk factors (notably smoking, hypertension and probably a history of migraine). Former users of COCs do not have an increased risk of ischaemic stroke. There is insufficient information to determine whether major differences in the risk of ischaemic stroke exist between products. Current users appear to have a modestly elevated risk of haemorrhagic stroke, mainly in women older than 35 years; former users do not. Data examining the risk of haemorrhagic stroke in current COC users with other risk factors are very sparse, as are those relating to the haemorrhagic stroke risk associated with particular COCs. Numerous studies have found, with remarkable consistency, an elevated risk of venous thromboembolism among current users of low estrogen dose COCs. The risk is substantially elevated among women with various inherited clotting factor defects. The effects in COC users with other risk factors for venous thrombosis tend to be less pronounced and more inconsistent. A number of studies have found higher relative risks among current users of low estrogen dose COCs containing desogestrel or gestodene, than among users of similar products containing levonorgestrel. A number of explanations, in terms of bias or confounding, have been proposed for these clinically small differences. At best, empirical evidence for these explanations, is weak. The risk of cardiovascular disease of any description is low in COC users. Women can minimise, and possibly eliminate entirely, their arterial risks by not smoking and by having their blood pressure checked before using a COC (in order to avoid its use if raised blood pressure is discovered). Users may decrease their venous thromboembolic risk by their choice of COC preparation although the effects will be modest. PMID- 10830254 TI - Comparative tolerability of chemotherapy regimens for germ cell cancer. AB - Germ cell tumours, even at an advanced stage, represent a unique model of malignant curable disease since >80% of patients are expected to be cured after appropriate therapy: surgery and radiotherapy in early stages, and chemotherapy and surgery in advanced stages. In advanced stages, serum tumour marker levels as well as extrapulmonary (brain, liver and bone) visceral metastases are the most important prognostic factors that affect treatment modalities. 'Gold standard' regimens for germ cell cancer currently include etoposide plus cisplatin with (BEP) or without (EP) bleomycin. In patients with good risk disease (90% cure rate), the optimal regimen of chemotherapy should combine the best efficacy and the least toxicity. As a result of randomised trials, 3 regimens can be currently recommended: (i) 4 cycles of EP; (ii) 4 cycles of BEP (with etoposide 350 mg/m2 per cycle); or (iii) 3 cycles of BEP (with etoposide 500 mg/m2 per cycle). In patients with poor risk disease, 4 cycles of BEP (with etoposide 500 mg/m2 per cycle) allow a disappointing cure rate of 50%. The long term toxicity of these regimens (gonadal toxicity and secondary malignancies) appears to be negligible and clearly does not challenge current standard strategies. PMID- 10830255 TI - A risk-benefit assessment of drugs used for neonatal chronic lung disease. AB - Improvements in neonatal intensive care have resulted in more extremely low birthweight babies surviving who are at risk of developing chronic lung disease. The preterm lung is vulnerable as it is both structurally immature and deficient in surfactant and antioxidant defences. Mechanical ventilation and high inspired oxygen concentrations are often necessary for preterm babies to survive but they can cause pulmonary inflammation which leads to lung damage. Abnormal healing in the presence of ongoing inflammation leads to airways remodelling which can result in protracted respiratory problems in these babies. A commonly used definition for chronic lung disease is the requirement for supplemental oxygen beyond 36 weeks' postconception. Many drugs that are commonly used for chronic lung disease have not been subjected to proper randomised controlled trials but are widely used on the basis of small studies showing short term benefits. They can be broadly divided into 2 groups. First, there are preventative drugs that are administered early to reduce oxygen toxicity and pulmonary inflammation. Secondly, there are those administered in established chronic lung disease, designed to reduce respiratory morbidity. Pulmonary inflammation in the neonate is reduced by systemic corticosteroids. Corticosteroid therapy within the first 2 weeks of life enables earlier extubation of preterm babies with subsequent reduced chronic lung disease and improved neonatal survival when given between 7 and 14 days. However, there is an increased risk of gastrointestinal haemorrhage, metabolic derangement, ventricular hypertrophy and potential effects on long term growth and brain development. Diuretics and inhaled bronchodilators improve pulmonary compliance and reduce oxygen requirements in established chronic lung disease but probably have little effect in reducing the incidence. In babies with established chronic lung disease, home oxygen therapy enables earlier discharge and prophylaxis against respiratory syncytial virus can reduce morbidity from bronchiolitis. All of the above therapies have adverse effects that need to be considered before initiating treatment. Recently, new drugs have become available which may be beneficial. These include inhaled nitric oxide for reduction of ventilation-perfusion mismatching, recombinant human superoxide dismutase for protection against oxidative stress and alpha-1 proteinase inhibitor which may reduce airways remodelling. At present these therapies are undergoing clinical trials. Exogenous surfactant is beneficial in respiratory distress syndrome and may reduce the risk of chronic lung disease but there have been no randomised controlled trials of its use in established chronic lung disease. Drugs which have been tried unsuccessfully include erythromycin, ambroxol and mast cell stabilisers. PMID- 10830256 TI - Benefits and risks of minocycline in rheumatoid arthritis. AB - Rheumatoid arthritis is a chronic inflammatory disease affecting about 1% of the adult population. The pathophysiology of rheumatoid arthritis remains incompletely understood. An infectious aetiology of the disease has long been postulated, but not proved. Despite insufficient evidence for the infectious nature of this disorder, several antibacterials, such as sulfa compounds, tetracyclines and rifampicin, have been investigated in the treatment of rheumatoid arthritis. In the last few years, minocycline, a semi-synthetic derivative of tetracycline, has been extensively studied as a therapeutic agent for rheumatoid arthritis. The antirheumatic effect of minocycline can be related to its immunomodulatory and anti-inflammatory, rather than to its antibacterial properties. Its efficacy in rheumatoid arthritis has been reported in 2 open trials and in 3 double-blind controlled studies. The first 2 double-blind studies, 1 in The Netherlands and 1 in the US, were performed in patients with advanced disease. Both studies showed a modest, but statistically significant improvement in the clinical parameters of disease activity and in the erythrocyte sedimentation rate in the minocycline-treated patients. The US study also reported that patients in the minocycline group developed fewer erosions than those in the placebo group. This finding supports the role of minocycline as a disease modifying agent. The common adverse effects of minocycline reported in these 2 studies included gastrointestinal adverse effects, dizziness, rash and headaches. Less common adverse effects were intracranial hypertension, pneumonitis, persistent skin and mucosal hyperpigmentation, lupus-like syndrome and acute hepatic injury. The third double-blind study enrolled only seropositive rheumatoid arthritis patients with early disease (less than 1 year duration), and showed very encouraging results of significant improvement in the disease activity parameters in the minocycline treated group of patients. The same authors later reported that about half of these patients were in or near remission after 3 years of follow up. No adverse effects were reported in this study. Summarising the data of these 3 double-blind studies, we may conclude that minocycline may be beneficial in patients with rheumatoid arthritis, especially when given early in the disease course or in patients with a mild disease. PMID- 10830257 TI - Sinorhizobium meliloti nfe (nodulation formation efficiency) genes exhibit temporal and spatial expression patterns similar to those of genes involved in symbiotic nitrogen fixation. AB - The nfe genes (nfeA, nfeB, and nfeD) are involved in the nodulation efficiency and competitiveness of the Sinorhizobium meliloti strain GR4 on alfalfa roots. The nfeA and nfeB genes are preceded by functional nif consensus sequences and NifA binding motifs. Here, we determined the temporal and spatial expression patterns of the nfe genes in symbiosis with alfalfa. Translational fusions of the nfe promoters with the gusA gene and reverse transcription-polymerase chain reaction analyses indicate that they are expressed and translated within mature nitrogen-fixing nodules and not during early steps of nodule development. Within the nodules the three nfe genes exhibit a spatial expression pattern similar to that of genes involved in symbiotic nitrogen fixation. We show that nfeB and nfeD genes are expressed not only from their own promoters but also from the upstream nfe promoter sequences. Furthermore, with the use of specific antibodies the NfeB and NfeD proteins were detected within the root nodule bacteroid fraction. Finally, NfeB was inmunolocalized in the bacteroid cell membrane whereas NfeD was detected in the bacteroid cytoplasm. PMID- 10830258 TI - A cluster of mutations disrupt the avirulence but not the virulence function of AvrPto. AB - avrPto in Pseudomonas syringae pv. tomato encodes an avirulence protein that triggers race-specific resistance in tomato plants carrying Pto. The AvrPto protein is secreted from P. syringae pv. tomato to plant cells through the type III secretion pathway and activates race-specific resistance by a direct interaction with the Pto protein. Here we report that avrPto enhances the virulence of P. syringae pv. tomato in a strain-dependent manner in tomato plants lacking Pto. To determine whether the virulence function can be structurally separated from the avirulence function, we examined the virulence activity of a group of AvrPto mutants that carry single amino acid substitutions and lack the avirulence activity on tomato plants. Three mutants that were clustered in the center of AvrPto exhibited virulence activity in tomato plants with or without Pto. The rest of the mutations abolished the virulence. The identification of these mutants suggested that the avirulence function of AvrPto can be structurally separated from the virulence function. PMID- 10830259 TI - Cell-to-cell movement of the 25K protein of potato virus X is regulated by three other viral proteins. AB - The 25K, 12K, and 8K proteins and coat protein (CP) of Potato virus X (PVX) are required for virus cell-to-cell movement. In this study, experiments were conducted to determine whether the PVX 25K protein moves cell to cell and to explore potential interactions between the PVX 25K, 12K, and 8K proteins and CP. The PVX 25K gene was fused to the green fluorescent protein (GFP) gene and inserted into plasmids adjacent to the cauliflower mosaic virus 35S promoter. These plasmids were introduced by biolistic bombardment to transgenic tobacco expressing the PVX 12K, 8K, and CP genes. The GFP:25K fused proteins moved cell to cell on nontransgenic tobacco and tobacco expressing either the 12K or 8K proteins. However, the GFP:25K proteins did not move on transgenic tobacco expressing the combined 12K/8K genes or the CP gene. Thus, movement of the PVX 25K protein through plasmodesmata may be regulated by interactions with other PVX proteins. PMID- 10830260 TI - Mesorhizobium loti increases root-specific expression of a calcium-binding protein homologue identified by promoter tagging in Lotus japonicus. AB - A promoter tagging program in the legume Lotus japonicus was initiated to identify plant genes involved in the nitrogen-fixing symbiosis between legumes and rhizobia. Seven transformed plant lines expressing the promoterless reporter gene uidA (beta-glucuronidase; GUS) specifically in roots and/or nodules were identified. Four of these expressed GUS in the roots only after inoculation with nodule-forming Mesorhizobium loti. In one line (T90), GUS activity was found in the root epidermis, including root hairs. During seedling growth, GUS expression gradually became focused in developing nodules and disappeared from root tissue. No GUS activity was detected when a non-nodulating mutant of M. loti was used to inoculate the plants. The T-DNA insertion in this plant line was located 1.3 kb upstream of a putative coding sequence with strong homology to calcium-binding proteins. Four motifs were identified, which were very similar to the "EF hands" in calmodulin-related proteins, each binding one Ca2+. We have named the gene LjCbp1 (calcium-binding protein). Northern (RNA) analyses showed that this gene is expressed specifically in roots of L. japonicus. Expression was reduced in roots inoculated with non-nodulating M. loti mutants and in progeny homozygous for the T-DNA insertion, suggesting a link between the T-DNA insertion and this gene. PMID- 10830261 TI - Temporal and spatial order of events during the induction of cortical cell divisions in white clover by Rhizobium leguminosarum bv. trifolii inoculation or localized cytokinin addition. AB - We examined the timing and location of several early root responses to Rhizobium leguminosarum bv. trifolii infection, compared with a localized addition of cytokinin in white clover, to study the role of cytokinin in early signaling during nodule initiation. Induction of ENOD40 expression by either rhizobia or cytokinin was similar in timing and location and occurred in nodule progenitor cells in the inner cortex. Inoculation of rhizobia in the mature root failed to induce ENOD40 expression and cortical cell divisions (ccd). Nitrate addition at levels repressing nodule formation inhibited ENOD40 induction by rhizobia but not by cytokinin. ENOD40 expression was not induced by auxin, an auxin transport inhibitor, or an ethylene precursor. In contrast to rhizobia, cytokinin addition was not sufficient to induce a modulation of the auxin flow, the induction of specific chalcone synthase genes, and the accumulation of fluorescent compounds associated with nodule initiation. However, cytokinin addition was sufficient for the localized induction of auxin-induced GH3 gene expression and the initiation of ccd. Our results suggest that rhizobia induce cytokinin-mediated events in parallel to changes in auxin-related responses during nodule initiation and support a role of ENOD40 in regulating ccd. We propose a model for the interactions of cytokinin with auxin, ENOD40, flavonoids, and nitrate during nodulation. PMID- 10830262 TI - High level activation of vitamin B1 biosynthesis genes in haustoria of the rust fungus Uromyces fabae. AB - In the rust fungus Uromyces fabae, the transition from the early stages of host plant invasion toward parasitic growth is accompanied by the activation of many genes (PIGs = in planta induced genes). Two of them, PIG1 (= THI1) and PIG4 (= THI2), were found to be highly transcribed in haustoria, and are homologous to genes involved in thiamine (vitamin B1) biosynthesis in yeast. Their functional identity was confirmed by complementation of Schizosaccharomyces pombe thiamine auxotrophic thi3 (nmt1) and thi2 (nmt2) mutants, respectively. In contrast to thiamine biosynthesis genes of other fungi that are completely suppressed by thiamine, THI1 and THI2 expression was not affected by the addition of thiamine to rust hyphae grown either in vitro or in planta. Immunoblot analysis revealed decreasing amounts of THI1p in extracts from spores, germlings, and in vitro grown infection structures with increasing time after inoculation. Immunofluorescence microscopy of rust-infected leaves detected high concentrations of THI1p in haustoria, and only low amounts in intercellular hyphae. In the sporulating mycelium, THI1p was found in the basal hyphae of the uredia, but not in the pedicels and only at very low levels in uredospores. These data indicate that the haustorium is an essential structure of the biotrophic rust mycelium not only for nutrient uptake but also for the biosynthesis of metabolites such as thiamine. PMID- 10830263 TI - Plants secrete substances that mimic bacterial N-acyl homoserine lactone signal activities and affect population density-dependent behaviors in associated bacteria. AB - In gram-negative bacteria, many important changes in gene expression and behavior are regulated in a population density-dependent fashion by N-acyl homoserine lactone (AHL) signal molecules. Exudates from pea (Pisum sativum) seedlings were found to contain several separable activities that mimicked AHL signals in well characterized bacterial reporter strains, stimulating AHL-regulated behaviors in some strains while inhibiting such behaviors in others. The chemical nature of the active mimic compounds is currently unknown, but all extracted differently into organic solvents than common bacterial AHLs. Various species of higher plants in addition to pea were found to secrete AHL mimic activities. The AHL signal-mimic compounds could prove to be important in determining the outcome of interactions between higher plants and a diversity of pathogenic, symbiotic, and saprophytic bacteria. PMID- 10830264 TI - Evidence for Agrobacterium-induced apoptosis in maize cells. AB - Agrobacterium spp. can genetically transform most dicotyledonous plant cells whereas many monocot species are recalcitrant to Agrobacterium-mediated transformation. One major obstacle is that co-cultivation of Agrobacterium spp. with plant tissues often results in cell death. Report here is that, in maize tissues, this process resembles apoptosis, with characteristic DNA cleavage into oligonucleosomal fragments and morphological changes. Two anti-apoptotic genes from baculovirus, p35 and iap, had the ability to prevent the onset of apoptosis triggered by Agrobacterium spp. in maize tissues. p35 is reported to act as a direct inhibitor of a certain class of proteases (caspase) whereas i.a.p. may act upstream to prevent their activation. This evidence raises the possibility that caspase-like proteases may also be involved in the apoptotic pathway in plant cells. PMID- 10830265 TI - Determination of the T-DNA transfer and the T-DNA integration frequencies upon cocultivation of Arabidopsis thaliana root explants. AB - Using the Cre/lox recombination system, we analyzed the extent to which T-DNA transfer to the plant cell and T-DNA integration into the plant genome determine the transformation and cotransformation frequencies of Arabidopsis root cells. Without selection for transformation competence, the stable transformation frequency of shoots obtained after cocultivation and regeneration on nonselective medium is below 0.5%. T-DNA transfer and expression occur in 5% of the shoots, indicating that the T-DNA integrates in less than 10% of the transiently expressing plant cells. A limited fraction of root cells, predominantly located at the wounded sites and in the pericycle, are competent for interaction with agrobacteria and the uptake of a T-DNA, as demonstrated by histochemical GUS staining. When selection for transformation competence is applied, the picture is completely different. Then, approximately 50% of the transformants show transient expression of a second, nonselected T-DNA and almost 50% of these cotransferred T DNAs are integrated into the plant genome. Our results indicate that both T-DNA transfer and T-DNA integration limit the transformation and cotransformation frequencies and that plant cell competence for transformation is based on these two factors. PMID- 10830266 TI - Disruption of a gene essential for sulfoquinovosyldiacylglycerol biosynthesis in Sinorhizobium meliloti has no detectable effect on root nodule symbiosis. AB - The sulfolipid sulfoquinovosyldiacylglycerol is commonly found in the thylakoid membranes of photosynthetic bacteria and plants. While there is a good correlation between the occurrence of sulfolipid and photosynthesis, a number of exceptions are known. Most recently, sulfoquinovosyldiacylglycerol was discovered in the non-photosynthetic, root nodule-forming bacterium Sinorhizobium meliloti. This discovery raised the questions of the phylogenetic origin of genes essential for the biosynthesis of this lipid in S. meliloti and of a function of sulfolipid in root nodule symbiosis. To begin to answer these questions, we isolated and inactivated the sqdB gene of S. meliloti. This gene and two other genes located directly 3' of sqdB are highly similar to the sqdB, sqdC, and sqdD genes known to be essential for sulfolipid biosynthesis in the photosynthetic, purple bacterium Rhodobacter sphaeroides. This observation confirms the close phylogenetic kinship between these two species. Furthermore, the reduced similarity of sqdB to the plant ortholog SQD1 of Arabidopsis thaliana does not support a previous sqd gene transfer from the plant as a consequence of close symbiosis. A sulfolipid deficient mutant of S. meliloti disrupted in sqdB is capable of inducing functional nodules and does not show an obvious disadvantage under different laboratory culture conditions. Thus far, no specific function can be assigned to bacterial sulfolipid, in either nodule-associated or free-living cells. S. meliloti contains a rich set of polar membrane lipids some of which, including sulfolipid, may become critical only under growth conditions that still need to be discovered. PMID- 10830267 TI - Genetic mapping of the Tsw locus for resistance to the Tospovirus Tomato spotted wilt virus in Capsicum spp. and its relationship to the Sw-5 gene for resistance to the same pathogen in tomato. AB - The Tsw gene conferring dominant resistance to the Tospovirus Tomato spotted wilt virus (TSWV) in Capsicum spp. has been tagged with a random amplified polymorphic DNA marker and mapped to the distal portion of chromosome 10. No mapped homologues of Sw-5, a phenotypically similar dominant TSWV resistance gene in tomato, map to this region in C. annuum, although a number of Sw-5 homologues are found at corresponding positions in pepper and tomato. The relationship between Tsw and Sw-5 was also examined through genetic studies of TSWV. The capacity of TSWV-A to overcome the Tsw gene in pepper and the Sw-5 gene in tomato maps to different TSWV genome segments. Therefore, despite phenotypic and genetic similarities of resistance in tomato and pepper, we infer that distinct viral gene products control the outcome of infection in plants carrying Sw-5 and Tsw, and that these loci do not appear to share a recent common evolutionary ancestor. PMID- 10830268 TI - The dual function in virulence and host range restriction of a gene isolated from the pPATH (Ehg) plasmid of Erwinia herbicola pv. gypsophilae. AB - The host range of the gall-forming bacterium Erwinia herbicola pv. gypsophilae (Ehg) is restricted to gypsophila whereas Erwinia herbicola pv. betae (Ehb) attacks beet as well as gypsophila. Both pathovars contain an indigenous plasmid (pPATH(Ehg or pPATH(Ehb)) that harbors pathogenicity genes, including the hrp gene cluster. A cosmid library of Ehg824-1 plasmid DNA was mobilized into Ehb4188 and the transconjugants were screened for pathogenicity on beet. One Ehb transconjugant harboring the cosmid pLA173 of pPATHEb induced a hypersensitive like response and abolished pathogenicity on beet. Transposon mutagenesis of an open reading frame (ORF) located on this cosmid eliminated its affect on pathogenicity. Marker exchange of this mutation into Ehg824-1 caused a substantial reduction in gall size on gypsophila and caused Ehg824-1 to extend its host range and incite galls on beet. The ORF (1.5 kb) was designated as pthG (pathogenicity gene on gypsophila). DNA sequence analysis of pthG revealed no significant homology to known genes in the data bank. Only remnants of the pthG sequences were identified on the pPATH of Ehb4188. The deduced protein lacked an N-terminal signal peptide but contained a short trans-membrane helix in its C terminus. The gene product, as determined by expression in Escherichia coli and Western blots (immunoblots), was a 56-kDa protein. PMID- 10830269 TI - The pre-symbiotic growth of arbuscular mycorrhizal fungi is induced by a branching factor partially purified from plant root exudates. AB - Arbuscular mycorrhizal (AM) symbiosis is an association between obligate biotrophic fungi and more than 80% of land plants. During the pre-symbiotic phase, the host plant releases critical metabolites necessary to trigger fungal growth and root colonization. We describe the isolation of a semipurified fraction from exudates of carrot hairy roots, highly active on germinating spores of Gigaspora gigantea, G. rosea, and G. margarita. This fraction, isolated on the basis of its activity on hyphal branching, contains a root factor (one or several molecules) that stimulates, directly or indirectly, G. gigantea nuclear division. We demonstrate the presence of this active factor in root exudates of all mycotrophic plant species tested (eight species) but not in those of nonhost plant species (four species). We negatively tested the hypothesis that it was a flavonoid or a compound synthesized via the flavonoid pathway. We propose that this root factor, yet to be chemically characterized, is a key plant signal for the development of AM fungi. PMID- 10830270 TI - Perspective: The insulin signaling system--a common link in the pathogenesis of type 2 diabetes. PMID- 10830271 TI - Perspective: Hexosamines and nutrient sensing. PMID- 10830272 TI - Perspective: Postnatal pancreatic beta cell growth. PMID- 10830273 TI - Persistent activation of phosphatidylinositol 3-kinase causes insulin resistance due to accelerated insulin-induced insulin receptor substrate-1 degradation in 3T3-L1 adipocytes. AB - Recently, we have reported that the overexpression of a membrane-targeted phosphatidylinositol (PI) 3-kinase (p110CAAX) stimulated p70S6 kinase, Akt, glucose transport, and Ras activation in the absence of insulin but inhibited insulin-stimulated glycogen synthase activation and MAP kinase phosphorylation in 3T3-L1 adipocytes. To investigate the mechanism of p110CAAX-induced cellular insulin resistance, we have now studied the effect of p110CAAX on insulin receptor substrate (IRS)-1 protein. Overexpression of p110CAAX alone decreased IRS-1 protein levels to 63+/-10% of control values. Insulin treatment led to an IRS-1 gel mobility shift (most likely caused by serine/threonine phosphorylation), with subsequent IRS-1 degradation. Moreover, insulin-induced IRS-1 degradation was enhanced by expression of p110CAAX (61+/-16% vs. 13+/-15% at 20 min, and 80+/-8% vs. 41+/-12% at 60 min, after insulin stimulation with or without p110CAAX expression, respectively). In accordance with the decreased IRS 1 protein, the insulin-stimulated association between IRS-1 and the p85 subunit of PI 3-kinase was also decreased in the p110CAAX-expressing cells, and IRS-1 associated PI 3-kinase activity was decreased despite the fact that total PI 3 kinase activity was increased. Five hours of wortmannin pretreatment inhibited both serine/threonine phosphorylation and degradation of IRS-1 protein. These results indicate that insulin treatment leads to serine/threonine phosphorylation of IRS-1, with subsequent IRS-1 degradation, through a PI 3-kinase-sensitive mechanism. Consistent with this, activated PI 3-kinase phosphorylates IRS-1 on serine/threonine residues, leading to IRS- 1 degradation. The similar finding was observed in IRS-2 as well as IRS-1. These results may also explain the cellular insulin-resistant state induced by chronic p110CAAX expression. PMID- 10830274 TI - Exendin-4 decelerates food intake, weight gain, and fat deposition in Zucker rats. AB - Exendin-4 is a 39 amino acid peptide produced in the salivary gland of the Gila monster lizard. It has a 53% amino acid homology to the incretin hormone glucagon like peptide-1 (GLP-1). Exendin-4 induces insulin release through activation of the GLP- 1 receptor but is a much more potent insulinotropic agent than GLP-1. Of critical importance for its potential use as a treatment for diabetes is its much longer biological effect in vivo. Previous studies involving once daily administration of exendin-4 over 13 weeks to db/db mice demonstrated that it lowers hemoglobin A1c (HbA1c), a marker of mean blood glucose levels. Food consumption in the treated animals dropped over the first 4 days and then increased to a level comparable with that of the untreated animals. In this study, we initially examined the effect of once daily injections (over 14 days) on the food consumption of Zucker fatty rats. We observed an immediate reduction in food intake which then leveled off(after 5 days) to match that of the untreated animals. Subsequently we injected the same animals twice daily (treatment period of 56 days in total) and observed a sustained reduction in food intake and weight-gain. This was matched by a reduction in the critical parameters of HbA1c, fasting blood glucose and plasma insulin. MRI imaging of the abdominal regions of the animals showed that initially only the amount of fat deposited in the sc region was reduced after 4 weeks exendin-4 treatment. At the 8-week time point there was a corresponding decrease in the amount of visceral fat deposition. The combination of appetite reduction, decreased fat deposition and an improvement in the parameters associated with glucose intolerance makes a case for the use of exendin-4 as a treatment for diabetes. PMID- 10830275 TI - Agouti-related protein-like immunoreactivity: characterization of release from hypothalamic tissue and presence in serum. AB - A novel RIA was used to examine the release of agouti-related protein-like immunoreactivity (AGRP-LI) from perfused rat hypothalamic tissue slices and to characterize AGRP-LI in rat serum. A continuous low level basal AGRP-LI release was observed from hypothalami of rats fed ad libitum before the rats were killed. Basal AGRP-LI release was 3-fold greater in rats fasted 48 h. In fasted animals leptin dose-dependently suppressed basal AGRP-LI release. In fed animals no change in basal AGRP-LI release was detected in response to 10(-6) M alpha-MSH, orexin B, melanin-concentrating hormone, or serotonin. HPLC analysis of AGRP-LI in rat serum identified a single peak that eluted in close proximity to synthetic AGRP (87-132) and mouse [Leu127Pro]AGRP and that was identical to the peak seen in hypothalamic and adrenal tissue extracts. The serum concentration of AGRP-LI in rats fed ad libitum was 0.865+/-0.323 nmol/liter (mean +/- SE). Food deprivation resulted in a slow, but statistically significant rise in serum immunoreactivity at 48 h [1.174+/-0.118 nmol/liter (mean +/- SE)]. Bilateral adrenalectomy did not change serum levels of AGRP-LI. These studies demonstrate that in the rat there are different levels of basal hypothalamic AGRP-LI release in fed and fasted states and that in the fasted rat this release can be profoundly suppressed by leptin. These studies also suggest that AGRP is present in the systemic circulation of rats. PMID- 10830276 TI - Blood to brain transfer of leptin in normal and interleukin-1beta-treated male rats. AB - To test the hypothesis that leptin was secreted from the brain into the blood of the rat, its concentration was measured in the superior sagittal sinus (SSS; which drains the cerebral cortex) and aortic blood of normal fasting male rats and rats that had been treated with iv or intracerebroventricular (icv) injections of interleukin-1beta (IL-1beta; 100 ng), a cytokine previously shown to induce peripheral leptin secretion. Plasma levels of leptin in SSS were slightly, but significantly, less than those in the aorta in control, saline injected rats (0.99+/-0.07 vs. 1.19+/-0.10 ng/ml; n = 15; P = 0.03) and in rats injected with human IL-1beta iv (1.56+/-0.12 vs. 1.92+/-0.15 ng/ml; n = 23; P = 0.004) or icv (1.38+/-0.11 vs. 1.57+/-0.12 ng/ml; n = 23; P = 0.008). IL-1beta by either the iv or icv route significantly increased leptin levels in the aorta [1.19+/-0.10 vs. 1.92+/-0.15 ng/ml (P = 0.0002) and 1.19+/-0.10 vs. 1.57+/-0.12 ng/ml (P = 0.022), respectively]. SSS levels of leptin were also raised after iv or icv injection (P = 0.0002 and P = 0.0053, respectively). These findings demonstrate a net uptake of leptin by the cerebral cortex from peripheral blood in both normal and IL-1beta-treated animals and show that peripheral blood levels of leptin are increased by IL-1beta whether administered icv or iv. PMID- 10830277 TI - Transcellular transport of leptin by the short leptin receptor isoform ObRa in Madin-Darby Canine Kidney cells. AB - Leptin is an adipocyte-derived hormone that acts in specific regions of the brain to regulate body weight and neuroendocrine function. The mechanism by which leptin enters the brain is unknown. We previously reported that rat brain microvessels, which constitute the blood-brain barrier, contain large amounts of messenger RNA encoding a short form of the leptin receptor (ObRa), suggesting that this site may be important for receptor-mediated transport of leptin into the brain. The purpose of this study was to determine whether ObRa is capable of transcellular transport of intact leptin. A transwell system in which Madin-Darby Canine Kidney (MDCK) cells stably expressing ObRa are grown in a monolayer was used to determine receptor distribution on apical or basolateral cell surfaces and the capacity for directional transport of 125I-leptin. Binding of 125I-leptin was greater on the apical vs. the basolateral cell surface and transport of 125I leptin occurred only in the apical to basolateral direction. 11% of transported radioactivity appearing in the basolateral chamber represented intact leptin as assessed by TCA precipitation analysis and by SDS-PAGE. Parental MDCK cells did not express leptin receptors and did not bind or transport 125I-leptin. Epidermal growth factor (EGF) binding and transport via endogenous EGF receptors in MDCK cells also was assessed. In contrast to leptin, specific binding of 125I-EGF occurred primarily on the basolateral cell surface and transport of 125I-EGF occurred predominantly in the basolateral to apical direction. These data show that ObRa is preferentially targeted to the apical cell membrane in MDCK cells and that leptin transport occurs, albeit at a low rate, in a unidirectional manner in the apical to basolateral direction. These findings may be relevant to the putative role of ObRa in receptor-mediated transport of leptin from the circulation into the brain. PMID- 10830278 TI - Overexpression of glutamine:fructose-6-phosphate amidotransferase in rat-1 fibroblasts enhances glucose-mediated glycogen accumulation via suppression of glycogen phosphorylase activity. AB - The hexosamine biosynthesis pathway (HBP) mediates many of the adverse effects of excess glucose. We have shown previously that glucose down-regulates basal and insulin-stimulated glycogen synthase (GS) activity. Overexpression of the rate limiting enzyme in the HBP, glutamine:fructose-6-phosphate amidotransferase (GFA), mimics these effects of high glucose and renders the cells more sensitive to glucose. Here we examine the role of the HBP in regulating cellular glycogen content. Glycogen content and glycogen phosphorylase (GP) activity were determined in Rat-1 fibroblasts that overexpress GFA. In both GFA and controls there was a dose-dependent increase in glycogen content (approximately 8-fold) in cells cultured in increasing glucose concentrations (1-20 mM). There was a shift to the left in the glucose dose-response curve for glycogen content in GFA cells (ED50 for glycogen content = 5.80+/-1.05 vs. 8.84+/-0.87 mM glucose, GFA vs. control). Inhibition of GFA reduced glycogen content by 28.4% in controls cultured in 20 mM glucose. In a dose-dependent manner, glucose resulted in a more than 35% decrease in GP activity in controls. GP activity in GFA cells was suppressed compared with that in controls, and there was no glucose-induced down regulation of GP activity. Glucosamine and uridine mimicked the effects of glucose on glycogen content and GP activity. However, chronic overexpression of GFA is a unique model of hexosamine excess, as culturing control cells in low dose glucosamine (0.1-0.25 mM) did not suppress GP activity and did not eliminate the glucose-mediated down-regulation of GP activity. We conclude that increased flux through the HBP results in enhanced glycogen accumulation due to suppression of GP activity. These results demonstrate that the HBP is an important regulator of cellular glucose metabolism and supports its role as a cellular glucose/satiety sensor. PMID- 10830279 TI - The in vivo and in vitro effects of exogenous leptin on ovulation in the rat. AB - Leptin, a hormonal product of the Lep gene, is expressed by adipocytes and is thought to play a role in regulating food intake and reproduction. The leptin protein has been localized in many reproductive tissues, including the ovary. Several publications indicate that the ovary is directly affected by leptin and that leptin may be a factor linking obesity and reproductive dysfunction. In this study, the effect of systemic leptin administration on ovulation in the rat ovary, both in vivo and in vitro, was investigated. Ip administration of leptin (30 microg at 3 hourly intervals for 15 h) to immature gonadotropin-primed rats caused a decline in ovulation in vivo, from 15.9+/-2.0 oocytes in the control animals to 5.3+/-1.6 oocytes in the leptin-treated animals (P < 0.001). Plasma progesterone and estradiol levels were analyzed immediately before ovulation, and neither was altered significantly in animals receiving the leptin treatment. Food consumption and body weight decreased following leptin treatment; however, a loss in body weight alone (pair-fed controls) was insufficient to explain the decrease in ovulation observed in the leptin-treated animals. In vitro perfusion of FSH primed whole ovaries showed that treatment with leptin in combination with LH significantly decreased ovulations from 5.7+/-1.6 per ovary perfused with LH alone to 1.3+/-0.6 in those with LH and 1 microg/ml leptin (P < 0.05). Progesterone and estradiol levels in the samples taken during the perfusion period were unaffected by leptin treatment. In summary, leptin administration resulted in fewer ovulations, both in vivo and in vitro, but did not influence steroid levels. Systemic leptin administration at these doses can therefore inhibit ovulation, a process that occurs through a direct effect on the ovary. PMID- 10830280 TI - Stat 5B, activated by insulin in a Jak-independent fashion, plays a role in glucokinase gene transcription. AB - Stat proteins are SH2 domain-containing transcription factors that are activated by various cytokines and growth factors. In a previous work, we have identified Stat 5B as a substrate of the insulin receptor based on yeast two-hybrid and mammalian cell transfection studies. In the present study, we have approached the biological relevance of the interaction between the insulin receptor and the transcription factor Stat 5B. Firstly, we show that both insulin and insulin-like growth factor I lead to tyrosine phosphorylation of Stat 5B, and this promotes binding of the transcription factor to the beta-casein promoter containing a Stat 5 binding site. Further, we demonstrate that insulin stimulates the transcriptional activity of Stat 5B. Activation of Stat 5B by insulin appears to be Jak2-independent, whereas Jak2 is required for GH-induced Stat 5B activation. Hence the pathway by which Stat 5B is activated by insulin is different from that used by GH. In addition, by using Jak1- and Tyk2-deficient cells we exclude the involvement of both Jak1 and Tyk2 in Stat 5B activation by insulin. Taken together, our results strengthen the notion that insulin receptor can directly activate Stat 5B. More importantly, we have identified a Stat 5 binding site in the human hepatic glucokinase promoter, and we show that insulin leads to a Stat 5B-dependent increase in transcription of a reporter gene carrying this promoter. These observations favor the idea that Stat 5B plays a role in mediating the expression of the glucokinase gene induced by insulin. As a whole, our results provide evidence for the occurrence of a newly identified circuit in insulin signaling in which the cell surface receptor is directly linked to nuclear events through a transcription factor. Further, we have revealed an insulin target gene whose expression is, at least in part, dependent on Stat 5B activation and/or binding. PMID- 10830281 TI - Long-chain acyl CoA regulation of protein kinase C and fatty acid potentiation of glucose-stimulated insulin secretion in clonal beta-cells. AB - Pancreatic beta-cells contain protein kinase C (PKC) isoforms that may play a role in insulin secretion. Activity of PKC classes (cPKC, nPKC, aPKC) and their regulation by acyl-CoA derivatives was examined in extracts of clonal pancreatic beta-cells (HIT) by protein phosphorylation. PKC classes were distinguished based on their previously defined cofactor requirements. Down-regulation of PKC by phorbol esters was confirmed by Western blotting and resulted in the complete loss of cPKC activity, partial loss of nPKC activity and preservation of aPKC activity and glucose-stimulated insulin secretion. aPKC activity was potentiated 4- to 8-fold by the CoA esters of myristate, palmitate, and oleate with a half maximal value of 3 microM. Both oleoyl- and myristol-CoA, but not palmitoyl-CoA, caused inhibition of nPKC activity. Oleoyl-CoA inhibited nPKC activity up to 75% with a half-maximal effect at 10 microM. This value was independent of the concentration of diacylglycerol used. The addition of exogenous oleate or palmitate potentiated glucose-stimulated insulin secretion 2-fold and was unaffected by PMA-induced down-regulation. Stimulation by glucose or glucose and oleate also increased the mass of PKC-zeta found in the particulate fraction. These data are consistent with increased cytosolic long-chain acylCoA-activating aPKC isoforms resulting in stimulation and/or potentiation of glucose-induced insulin secretion. PMID- 10830282 TI - Hexosamines stimulate leptin production in transgenic mice. AB - Hexosamine flux has been shown to mediate aspects of nutrient sensing in insulin sensitive tissues and has been hypothesized to represent a satiety signal that results in shunting of fuel toward storage as fat. It has been recently reported that in vitro treatment of fat and muscle cells with hexosamines and acute glucosamine infusion in intact rats stimulate leptin secretion. In order to investigate the effects of chronic, physiologic increases in hexosamine flux on leptin we have examined leptin mRNA and serum leptin in mice overexpressing the rate-limiting enzyme for hexosamine synthesis, GFA, in muscle and fat. Increased levels of UDP-N-acetylglucosamine, the principal end-product of the hexosamine pathway were seen in transgenic fat, consistent with the overexpression of GFA. After overnight fasting, the transgenic mice were hyperleptinemic compared to littermate controls (4.5+/-0.5 ng/ml in transgenic, 2.8+/-0.2 in control, p = 0.005) despite equal body weights. In the random-fed state, the leptin levels of control mice increased to 4.1+/-0.5 ng/ml (p = 0.01) whereas the leptin levels in the transgenics did not increase any further (3.7+/-0.4 ng/ml). Leptin mRNA levels were also increased in transgenic fat (2.7+/-0.6 in transgenic compared to 0.8+/-0.2 in control, arbitrary units normalized to actin, p < 0.007). Despite increased leptin, the transgenic animals did not have lower body fat content. We conclude that hexosamine flux in fat regulates leptin synthesis and secretion. PMID- 10830283 TI - Cytokines induce both necrosis and apoptosis via a common Bcl-2-inhibitable pathway in rat insulin-producing cells. AB - The presence of activated macrophages within pancreatic islets in insulin dependent diabetes mellitus suggests an involvement of beta-cell death by necrosis. The aim of this study was to investigate the frequencies and mechanisms of cytokine-induced beta-cell apoptosis and necrosis and the possible protection mediated by the antiapoptotic gene bcl-2. A combination of interleukin-1beta, interferon-gamma, and tumor necrosis factor-alpha increased both necrosis (17% of cells) and apoptosis (5% of cells) in isolated whole rat islets, as determined by vital staining and fluorescence microscopy. Hyperexpression of Bcl-2, achieved by stable transfection using a multicopy viral vector containing a bcl-2 complementary DNA in rat insulin-producing RINm5F cells, counteracted both apoptosis and necrosis. Cytokine-induced cleavage of the caspase-3 substrate poly(ADP-ribose) polymerase (which, in other cell types, may occur downstream or independently of a Bcl-2-preventable mitochondrial permeability transition) was observed in control- but neither in bcl-2-transfected cells nor in the presence of the iNOS inhibitor N(G)-methyl-L-arginine. Tumor necrosis factor-alpha alone did not clearly induce cell death or poly(ADP-ribose) polymerase-cleavage. These findings suggest that cytokines induce both necrosis and apoptosis in insulin producing cells via a common Bcl-2-preventable nitric oxide-dependent pathway, which may involve mitochondrial permeability transition. The necrosis:apoptosis ratio might be increased by a relative lack of caspase activity. PMID- 10830284 TI - The relationship between pulsatile secretion and calcium dynamics in single, living gonadotropin-releasing hormone neurons. AB - It is well established that pulsatile release of GnRH regulates the reproductive axis, but little is known about the mechanisms underlying this pulsatility. Recent findings that GT1 cells, a line derived from the mouse embryonic hypothalamus, release GnRH in a pulsatile manner indicates that this rhythmic activity is an intrinsic property of GnRH neurons. In several attempts to uncover the intracellular basis for this pulsatile phenomenon, it was revealed that intracellular calcium concentrations change in a rhythmic fashion in GnRH neurons and that cellular depolarization, which triggers a secretory event, is associated with profound calcium changes in the cells. These findings raised the intriguing possibility that periodic alterations in intracellular calcium concentrations may underlie the phenomenon of pulsatile secretion in GnRH neurons. To address this, we first adapted the use of FM1-43 fluorescence to monitor changes of secretion in individual GT1-7 cells and then combined this approach with simultaneous measurement ofintracellular free calcium ([Ca2+]i, fura 2 method). In initial validation experiments, we found that stimulation of exocytosis with K+ (75 mM) or N-methyl-D-aspartate (NMDA, 100 microM) predictably evoked dynamic increases of both FM1-43 and fura 2 fluorescence. Later measurement of calcium dynamics and exocytotic activity in unstimulated cells revealed that [Ca2+]i underwent transitions from quiescence to high oscillatory behavior, and that these shifts were frequently associated with exocytotic events. Moreover, these calcium oscillatory transitions and associated changes in secretory activity occurred synchronously among most adjacent cells and at a frequency similar to that reported for pulsatile release of GnRH by entire cultures of GnRH neurons. Taken together, these results indicate that the intrinsic secretory pulsatility of GnRH neurons appears to be a consequence of coordinated, periodic changes in the pattern of calcium oscillations within individual cells. PMID- 10830285 TI - Identification of an immunodominant region recognized by human autoantibodies in a three-dimensional model of thyroid peroxidase. AB - Autoimmune thyroid diseases (AITD) are characterized by the presence of autoantibodies to thyroid peroxidase (TPO). This response is dominated by autoantibodies to two conformational determinants, termed A and B, that have been defined by monoclonal antibodies but whose structures and location within TPO are unknown. We have modeled the three-dimensional structure of the extracellular region of TPO, raised antisera to prominent surface structures, and identified an epitope that we show to be a critical part of the B determinant. Antibodies to this epitope inhibit the binding to TPO of human autoantibodies in virtually all serum samples from 65 patients with AITD that were tested. This first description of a model of the three-dimensional structure and location of a major autoantigenic determinant within the TPO molecule may provide structural clues for identifying causative agents or developing novel therapeutic strategies. PMID- 10830286 TI - Prolactin induces apoptosis in the penultimate spermatogonial stage of the testes in Japanese red-bellied newt (Cynops pyrrhogaster). AB - Cell death is a common feature during spermatogenesis and, in some seasonal breeding animals, is often observed at the transition stage from spermatogonia to spermatocytes. In the Japanese red-bellied newt, we have previously shown that this cell death is caused by the elevated titer of plasma PRL that occurs after animals are transferred to low temperature, suggesting that cell death causes the cessation of spermatocytogenesis from late autumn to early spring. In the present report, first we show that the injection of PRL into newts causes apoptosis in spermatogonia after the sixth mitotic division, the penultimate one before spermatogonia normally enter meiosis. Second, we demonstrate in organ cultures of testes fragments that PRL acts directly on the testes. Third, we show that the action by PRL is inhibited by FSH dose dependently. PMID- 10830287 TI - Identification of ADAM 31: a protein expressed in Leydig cells and specialized epithelia. AB - A family of proteins containing a disintegrin and metalloproteinase domain (ADAMs) has been identified recently. Here, we report the identification of a novel member of the ADAM protein family from mouse. This protein is designated ADAM 31. The complementary DNA sequence of ADAM 31 predicts a transmembrane protein with metalloproteinase, disintegrin, cysteine-rich, and cytoplasmic domains. Messenger RNA encoding ADAM 31 was most abundant in testes, but was also detected in many other tissues. More significantly, the antibodies raised against ADAM 31 reveal that the protein has a unique and restricted expression pattern. ADAM 31 is expressed in Leydig cells of the testes, but unlike many other ADAMs, it is not found on developing sperm. Furthermore, ADAM 31 is highly expressed on four types of specialized epithelia: the cauda epididymidis, the vas deferens, the convoluted tubules of the kidney, and the parietal cells of the stomach. PMID- 10830288 TI - Evaluation of keratinocyte proliferation and differentiation in vitamin D receptor knockout mice. AB - The biological effects of 1,25-dihydroxyvitamin D3 are mediated by a nuclear receptor, the vitamin D receptor (VDR). Targeted ablation of the VDR in mice results in hypocalcemia, hypophosphatemia, hyperparathyroidism, rickets, osteomalacia, and alopecia. Normalization of mineral ion homeostasis prevents these abnormalities with the exception of the alopecia. Because 1,25(OH)2D3 has been shown to play a role in keratinocyte proliferation and differentiation, we undertook studies in primary keratinocytes and skin isolated from VDR null mice to determine if a keratinocyte abnormality could explain the alopecia observed. The basal proliferation rate of the VDR null and wild-type keratinocytes was identical both under proliferating and differentiating conditions. Assessment of in vivo keratinocyte proliferation at 4 days of age confirmed that VDR ablation did not have a significant effect. There was no difference in the basal expression of markers of keratinocyte differentiation (keratin 1, involucrin, and loricrin) in the keratinocytes isolated from VDR-ablated mice when compared with those isolated from control littermates. Similarly, in vivo expression of these genes was not altered at 4 days of age. When anagen was induced by depilation at 18 days of age, the VDR null mice had a profound impairment in initiation of the hair cycle. These data suggest that the alopecia in the VDR null mice is not attributable to an intrinsic defect in keratinocyte proliferation or differentiation, but rather to an abnormality in initiation of the hair cycle. PMID- 10830289 TI - A 278 bp region just upstream of the human CYP19 (aromatase) gene mediates ovary specific expression in transgenic mice. AB - In humans, the CYP19 gene, which encodes aromatase P450, is expressed in a number of tissues including gonads, adipose, bone, and placenta. The 5'-untranslated regions (UTR) of CYP19 mRNA transcripts in these tissues are encoded by different tissue-specific first exons, which are alternatively spliced onto a common site just upstream of the start site of translation in exon II. In ovary, the 5'-UTR of CYP19 transcripts is encoded by exon IIa, which lies just upstream of exon II. To map genomic sequences required for ovary-specific CYP19 expression, fusion genes containing 2700, 278 and 43 bp of exon IIa 5'-flanking DNA linked to the human growth hormone (hGH) gene, as reporter, were introduced into mice. CYP19(IIa)-2700:hGH and CYP19(IIa)-278:hGH transgenes were expressed at high levels in ovaries of transgenic mice, whereas, ovarian expression of CYP19(IIa) 43:hGH was undetectable. In the majority of mice carrying the CYP19(IIa)-2700:hGH and CYP19(IIa)-278:hGH fusion genes, transgene expression was ovary-specific, indicating that genomic sequences within 278 bp region mediate ovary-specific expression. Expression of CYP19(IIa)-2700:hGH and CYP19(IIa)-278:hGH fusion genes was detected in ovarian granulosa and luteal cells, as well as in luteinized interstitial cells in mice during the estrous cycle. The most intense hGH immunostaining was observed in corpora lutea. This pattern of transgene expression is similar to that of aromatase in women where both granulosa and luteal cells express aromatase during the menstrual cycle. These findings suggest that transgenic mice provide an excellent model for analyzing genomic regions that mediate ovary-specific expression of the human CYP19 gene. PMID- 10830290 TI - Knockout of the murine prostaglandin EP2 receptor impairs osteoclastogenesis in vitro. AB - Prostaglandin E2 (PGE2) stimulates the formation of osteoclast-like tartrate resistant acid phosphatase-positive multinucleated cells (TRAP + MNC) in vitro. This effect likely results from stimulation of adenylyl cyclase, which is mediated by two PGE2 receptors, designated EP2 and EP4. We used cells from mice in which the EP2 receptor had been disrupted to test its role in the formation of TRAP + MNC. EP2 heterozygous (+/-) mice in a C57BL/6 x 129/SvEv background were bred to produce homozygous null (EP2 -/-) and wild-type (EP2 +/+) mice. PGE2, PTH, or 1,25 dihydroxyvitamin D increased TRAP+ MNC in 7-day cultures of bone marrow cells from EP2 +/+ mice. In cultures from EP2 -/- animals, responses to PGE2, PTH, and 1,25 dihydroxyvitamin D were reduced by 86%, 58%, and 50%, respectively. A selective EP4 receptor antagonist (EP4RA) further inhibited TRAP+ MNC formation in both EP2 +/+ and EP2 -/- cultures. In cocultures of spleen and calvarial osteoblastic cells, the response to PGE2 or PTH was reduced by 92% or 85% when both osteoblastic cells and spleen cells were from EP2 -/- mice, by 88% or 68% when only osteoblastic cells were from EP2 -/- mice and by 58% or 35% when only spleen cells were from EP2 -/- mice. PGE2 increased receptor activator of nuclear factor (NF)-kappaB ligand (RANKL) messenger RNA expression in osteoblastic and bone marrow cell cultures from EP2 +/+ mice 2-fold but had little effect on cells from EP2 -/- mice. Spleen cells cultured with RANKL and macrophage colony stimulating factor produced TRAP+ MNC. PGE2 increased the number of TRAP+ MNC in spleen cell cultures from EP2 +/+ mice but not in cultures from EP2 -/- mice. EP4RA had no effect on the PGE2 response in spleen cell cultures. PGE2 decreased the expression of messenger RNA for granulocyte macrophage colony stimulating factor in spleen cell cultures from EP2 +/+ mice but had little effect on cells from EP2 -/- mice. These data demonstrate that the prostaglandin EP2 receptor plays a role in the formation of osteoclast-like cells in vitro. A major defect in EP2 -/- mice appears to be in the capacity of osteoblastic cells to stimulate osteoclast formation. In addition, there appears to be a defect in the response of cells of the osteoclastic lineage to PGE2 in EP2 -/- mice. PMID- 10830291 TI - Vascular endothelial growth factor and transforming growth factor-beta1 regulate the expression of insulin-like growth factor-binding protein-3, -4, and -5 in large vessel endothelial cells. AB - We investigated the effect of diabetes-associated growth factors on the expression of insulin-like growth factor-I (IGF-I) and IGF-binding proteins (IGFBPs) in cultured endothelial cells from bovine aorta. Gene expression was measured by solution hybridization, and proteins were measured by enzyme-linked immunosorbent assay, RIA, or Western blot. The cells expressed messenger RNA (mRNA) for IGFBP-2 through -6 and IGFBP-2 through -5 proteins were detected in conditioned medium. Vascular endothelial growth factor inhibited IGFBP-3 mRNA (P < 0.01) and protein expression and increased IGFBP-5 mRNA (P < 0.001) and protein. Transforming growth factor-beta1 inhibited IGFBP-3 (P < 0.01), IGFBP-4 (P < 0.01), and IGF-I mRNA expression, whereas at the protein level only IGFBP-3 was significantly decreased. IGF-I, insulin, or angiotensin II did not affect IGF I or IGFBP mRNA expression. At the protein level, IGF-I clearly increased IGFBP-5 levels in conditioned medium. In conclusion, vascular endothelial growth factor and transforming growth factor-beta1 regulate IGFBP expression in bovine aortic endothelial cells. These observations provide a new aspect of regulation for the IGF-system in macrovascular endothelium, with possible implications for subendothelial smooth muscle cells and development of diabetic angiopathy. PMID- 10830292 TI - Transforming growth factor-beta receptor types I and II in cultured porcine leydig cells: expression and hormonal regulation. AB - The steroidogenic activity of testicular Leydig cells is controlled both by the pituitary hormone (LH) and by growth factors such as transforming growth factor beta peptides (TGFbeta1, -2, and -3; inhibin/activin; and anti-Mullerian hormone). By using primary cultures of porcine Leydig cells as a model, the aim of the study was to identify and characterize the TGFbeta receptors and to study their regulation by LH/hCG. TGFbeta receptors have been identified and characterized through three different approaches, including cross-linking experiments and Western and Northern blotting analyses. In cross-linking experiments, labeled TGFbeta was shown to bind to three different molecular species of 300, 80, and 53 kDa, which may correspond to the protein betaglycan (also known as TGFbeta type III receptor) and TGFbeta type II and I receptors (TGFbetaRII and TGFbetaRI), respectively. The presence of TGFbetaRI and -RII was further demonstrated by Western blotting analysis using specific polyclonal antibodies. Finally, the expression of betaglycan, TGFbetaRII, and TGFbetaRI messenger RNAs, was confirmed by Northern blotting analysis, as shown by the presence of 6.4-, 4.6-, and 5.8-kb messenger RNAs, respectively. By using a RT PCR approach, the mediators of the TGFbeta signal, Smads 1-7, were also detected in cultured Leydig cells. TGFbetaRI and TGFbetaRII protein levels were enhanced by hCG/LH in a dose-dependent (maximal effect with 0.3 ng/ml hCG) and time dependent (maximal effect observed after 48 h of hCG treatment) manner. Furthermore, to determine whether the stimulatory effect of LH/hCG was mediated by testosterone, use was made of aminogluthetimide, an inhibitor of cytochrome P450scc. The inhibition oftestosterone formation did not affect the stimulatory effect of LH/hCG on TGFbetaRI and -RII levels, suggesting that the gonadotropin action is not mediated by the steroid hormone. Together, the present findings demonstrate that the TGFbeta receptors are expressed and are under hormonal (gonadotropin) control in cultured porcine Leydig cells. PMID- 10830293 TI - Mechanisms of fibroblast growth factor-2 modulation of vascular endothelial growth factor expression by osteoblastic cells. AB - Normal bone growth and repair is dependent on angiogenesis. Fibroblast growth factor-2 (FGF-2), vascular endothelial growth factor (VEGF), and transforming growth factor-beta (TGFbeta) have all been implicated in the related processes of angiogenesis, growth, development, and repair. The purpose of this study was to investigate the relationships between FGF-2 and both VEGF and TGFbeta in nonimmortalized and clonal osteoblastic cells. Northern blot analysis revealed 6 fold peak increases in VEGF mRNA at 6 h in fetal rat calvarial cells and MC3T3-E1 osteoblastic cells after stimulation with FGF-2. Actinomycin D inhibited these increases in VEGF mRNA, whereas cycloheximide did not. The stability ofVEGF mRNA was not increased after FGF-2 treatment. Furthermore, FGF-2 induced dose dependent increases in VEGF protein levels (P < 0.01). Although in MC3T3-E1 cells, TGFbeta1 stimulates a 6-fold peak increase in VEGF mRNA after 3 h of stimulation, we found that both TGFbeta2 and TGFbeta3 yielded 2- to 3-fold peak increases in VEGF mRNA levels noted after 6 h of stimulation. Similarly, both TGFbeta2 and TGFbeta3 dose dependently increased VEGF protein production. To determine whether FGF-2-induced increases in VEGF mRNA may have occurred independently of TGFbeta, we disrupted TGFbeta signal transduction (using adenovirus encoding a truncated form of TGFbeta receptor II), which attenuated TGFbeta1 induction of VEGF mRNA, but did not impede FGF-2 induction ofVEGF mRNA. In summary, FGF-2-induced VEGF expression by osteoblastic cells is a dose dependent event that may be independent of concomitant FGF-2-induced modulation of TGFbeta activity. PMID- 10830294 TI - High plasma growth hormone (GH) levels inhibit expression of GH secretagogue receptor messenger ribonucleic acid levels in the rat pituitary. AB - Synthetic GH secretagogues (GHSs) act via a receptor (GHS-R) distinct from that of GH-releasing hormone. The GHS-R has been cloned from the pituitary and is expressed not only in the pituitary but also in specific areas of the brain, including the hypothalamus. Recent studies suggest that hypothalamic GHS-R expression is regulated by GH. This study was designed to investigate whether pituitary GHS-R expression is modulated by GH. Female Wistar-Furth rats were injected sc with either saline (control) or GC tumor cells (GC) that secrete rat GH. The tumors were allowed to develop for 1-4 weeks. At weeks 1-4, control (n = 4-8) and GC rats (n = 3-8) were killed. Pituitary GHS-R messenger RNA (mRNA) was measured by a quantitative competitive PCR assay. The endogenous GHS-R mRNA levels were measured by determining the amount of competitive template RNA required to produce equimolar amounts of native and competitive template PCR products. The mean log plasma GH levels were significantly greater in the GC rat group than in the control group at weeks 2, 3, and 4. At these times, the mean log pituitary GHS-R mRNA contents were significantly lower in the GC rat group than in the control group. No relationship could be established between log estradiol levels and GHS-R levels. These data indicate that pituitary GHS-R expression is modulated by GH. PMID- 10830295 TI - Thyrotropin modulates interferon-gamma-mediated intercellular adhesion molecule-1 gene expression by inhibiting Janus kinase-1 and signal transducer and activator of transcription-1 activation in thyroid cells. AB - TSH is known as an important hormone that plays the major role not only in the maintenance of normal physiology but also in the regulation of immunomodulatory gene expression in thyrocytes. The adhesion molecule intercellular adhesion molecule-1 (ICAM-1) was identified as one of the proteins that are abnormally expressed in the thyroid gland during autoimmune thyroid diseases. In this study we found that TSH inhibits interferon-gamma (IFNgamma)-mediated expression of the ICAM-1 gene, and we investigated the involved mechanisms in rat FRTL-5 thyroid cells. After exposure to IFNgamma, ICAM-1 expression is positively regulated at the level of transcription. This effect occurs via the IFNgamma-activated site (GAS) element in the ICAM-1 promoter as a consequence of the activation of STAT1 (signal transducer and activator of transcription-1), but not of STAT3. On the other hand, after exposure to TSH plus IFNgamma, ICAM-1 transcription is negatively modulated. We found that this inhibitory effect of TSH also occurs via the GAS element. Electrophoretic mobility shift assays confirmed that the IFNgamma-induced DNA-binding activities of STAT1 were reduced by TSH. Furthermore, our results showed that the inhibitory effect of TSH on IFNgamma signaling is caused by inhibition of tyrosine phosphorylation on STAT1, Janus kinase-1 (Jak1), and IFNgamma receptor a, but not Jak2. In conclusion, we have identified a novel mechanism in which TSH modulates the IFNgamma-mediated Jak/STAT signaling pathway through the inhibition of Jak1 and STAT1. PMID- 10830296 TI - Adrenomedullin enhances cell proliferation and deoxyribonucleic acid synthesis in rat adrenal zona glomerulosa: receptor subtype involved and signaling mechanism. AB - The effect of adrenomedullin (ADM) on the proliferative activity of the rat adrenal cortex has been investigated in vivo, using an in situ perfusion technique of the intact left gland. ADM and other chemicals were dissolved in the perfusion medium, and the perfusion was continued for 180 min. ADM infusion concentration dependently increased the mitotic index and [3H]thymidine incorporation into DNA in the zona glomerulosa (ZG; the maximal effective concentration was 10(-8) M), but not in inner adrenocortical layers, where basal proliferative activity was negligible. The effect of 10(-8) M ADM was equipotently counteracted by both the calcitonin gene-related peptide (CGRP) type 1 receptor antagonist CGRP-(8-37) and ADM-(22-52). The adenylate cyclase inhibitor SQ-22536 (10(-4) M), the cAMP blocker Rp-cAMP-S (10(-3) M), and the protein kinase A inhibitor H-89 (10(-5) M), although counteracting the ZG proliferogenic action of 10(-9) M ACTH, did not affect the 10(-8) M ADM-elicited increase in ZG DNA synthesis. Similar results were obtained using the phospholipase C inhibitor U-73122 (10(-5) M), the inositol-1,4,5-trisphosphate antagonist D,L-myo-inositol-1,4,5-trisphosphothiate (10(-4) M), and the protein kinase C inhibitor calphostin C (10(-5) M), which, however, significantly inhibited the ZG proliferogenic effect of 10(-9) M angiotensin II. The growth promoting action of 10(-8) M ADM was not affected by the phospholipase A2 inhibitor AACOCF3 (10(-5) M), the cyclooxygenase (COX) inhibitor indomethacin (10(-5) M), or the mixed COX/lipoxygenase inhibitor phenidone (10(-5) M). In contrast, the ZG proliferogenic effect of 10(-8) M ADM was abolished by either the tyrosine kinase (TK) inhibitor tyrphostin-23 (10(-5) M) or the mitogen activated protein kinase (MAPK) antagonists PD-98059 and U0216 (10(-4) M). ADM (10(-8) M) stimulated TK and p42/p44 MAPK activity in dispersed ZG, but not ZF, cells, and the effect was reversed by either 10(-6) M CGRP-(8-37) and ADM-(22-52) or preincubation with 10(-5) M tyrphostin-23. Collectively, our findings indicate that 1) ADM stimulates cell proliferation in the rat ZG, through CGRP-(8-37)- and ADM-(22-52)-sensitive receptors, probably of the CGRP1 subtype; and 2) the mitogenic effect of ADM is mediated by activation of the TK-MAPK cascade, without any involvement of the adenylate cyclase/protein kinase A-, phospholipase C/protein kinase C-, and COX- or lipoxygenase-dependent signaling pathways. PMID- 10830297 TI - Immortalized luteinizing hormone-releasing hormone neurons show a different migratory activity in vitro. AB - The development of two cell lines (GT1 and GN) of immortalized LHRH neurons has allowed an accurate study of the mechanisms controlling the synthesis and the secretion of LHRH. These cell lines, obtained in mice by genetic targeted tumorigenesis, retain many of the phenotypic characteristics of LHRH neurons. Of interest, GT1 cells derive from an hypothalamic tumor, whereas GN cells were obtained from a tumor localized in the olfactory bulb. The different origin of these cell lines lead to hypothesize that they might represent hypothalamic postmigratory neurons (GT1 cells), or LHRH neurons blocked at an early stage of their migration (GN cells). Using different experimental procedures, we found that the two cell subclones GT1-7 and GN11 express a different morphology and migratory behavior in vitro. In particular, we found that GN11 cells, but not GT1 7 cells, show the morphological shape of migrating neurons. When analyzing the spontaneous motility we found that only GN11 cells express a high capacity of migrating in a matrix of collagen gel. Moreover, in a chemomigratory assay GN11 cells did show a significant response to the chemotactic stimulus represented by the FBS. On the contrary, GT1-7 cells show very low spontaneous motility and appear insensitive to the FBS stimulus. These results suggest that the simultaneous use of the GT1-7/GN11 cells may represent an experimental tool for screening the factors possibly involved in the control of the migratory processes of LHRH neurons in normal and in pathological conditions, such as those due to their impaired migration, like it happens in Kallmann's syndrome. PMID- 10830298 TI - Growth hormone-releasing hormone stimulates mitogen-activated protein kinase. AB - GH-releasing hormone (GHRH) can induce proliferation of somatotroph cells. The pathway involving adenylyl cyclase/cAMP/protein kinase A pathway in its target cells seems to be important for this action, or at least it is deregulated in some somatotroph pituitary adenomas. We studied in this work whether GHRH can also stimulate mitogen-activated protein (MAP) kinase. GHRH can activate MAP kinase both in pituitary cells and in a cell line overexpressing the GHRH receptor. Although both protein kinase A and protein kinase C could activate MAP kinase in the CHO cell line studied, neither protein kinase A nor protein kinase C appears to be required for GHRH activation of MAP kinase in this system. However, sequestration of the betagamma-subunits of the G protein coupled to the receptor inhibits MAP kinase activation mediated by GHRH. This pathway also involves p21ras and a phosphatidylinositol 3-kinase, probably phosphatidylinositol 3-kinase-gamma. Despite the involvement of p21ras, the protein kinase Raf-1 is not hyperphosphorylated in response to GHRH, contrary to what usually occurs when the Ras-Raf-MAP kinase pathway is activated. In summary, this work describes for the first time the activation of MAP kinase by GHRH and outlines a path for this activation that is different from the cAMP-dependent mechanism that has been traditionally described as mediating the mitogenic actions of GHRH. PMID- 10830299 TI - Antagonists of growth hormone-releasing hormone and vasoactive intestinal peptide inhibit tumor proliferation by different mechanisms: evidence from in vitro studies on human prostatic and pancreatic cancers. AB - Antagonists of GH-releasing hormone (GHRH) and vasoactive intestinal peptide (VIP) inhibit the proliferation of various tumors in vitro and in vivo, but a comparison of their antitumor effects and mechanisms of action has not been reported to date. We recently synthesized and characterized a series of analogs, some of which are primarily GHRH antagonists (JV-1-36, JV-1-38, and JV-1-42), whereas others are more selective for VIP receptors (VPAC-R; JV-1-50, JV-1-51, JV 1-52, and JV-1-53). LNCaP human prostatic cancer cells express VPAC-R, with predominant subtype 1 determined by RT-PCR. Our studies show that GHRH antagonists significantly inhibit the proliferation of both VPAC-R positive LNCaP cells (P < 0.001) and VPAC-R negative MiaPaCa-2 human pancreatic cancer cells cultured in vitro (P < 0.05 to P < 0.001). Growth inhibition of LNCaP cells is accompanied by a proportional reduction in prostate-specific antigen (PSA) secretion (P < 0.001). In a superfusion system, the inhibitory activities of the analogs on the rate of VIP and GHRH-induced PSA secretion correlate well with their VPAC-R binding affinities to LNCaP cell membranes. Antagonists more selective for VPAC-R display a stronger inhibition of inducible PSA release than GHRH antagonists, but have smaller effects or no effects on proliferation and PSA secretion in culture. Collectively, our findings demonstrate that the antiproliferative activity of the analogs on cancer cells is not correlated to their VPAC-R antagonistic potencies. Because GHRH antagonists inhibit the proliferation of LNCaP cells more powerfully than VPAC-R antagonists and also suppress the growth ofVPAC-R-negative MiaPaCa-2 cells, it can be concluded that their antiproliferative effect is exerted through a mechanism independent of VPAC R. PMID- 10830300 TI - Evidence for a functional association between phosphatidylinositol 3-kinase and c src in the spreading response of osteoclasts to colony-stimulating factor-1. AB - Osteoclasts are bone-resorbing cells whose normal function depends in part upon their ability to migrate over the bone surface to initiate new sites of bone resorption. The growth factor/cytokine, colony-stimulating factor-1 (CSF-1), potently stimulates osteoclast motility, in a c-src-dependent fashion. The intracellular signaling molecules that participate with c-src in CSF-1-induced remodeling of the osteoclast cytoskeleton have not been identified. Here we demonstrate, using the inhibitors wortmannin and LY294002, that activation of phosphatidylinositol 3-kinase (PI3-K) is required for CSF-1-induced spreading in osteoclasts. After CSF-1 treatment of osteoclast-like cells, PI3-K activity associated with the CSF-1 receptor c-fms, is increased, and the 85-kDa regulatory subunit of PI3-K and c-src coimmunoprecipitate. CSF-1 induces redistribution of PI3-K to the periphery of the cell. The association between p85 and c-src is due in part to a direct interaction between the proline-rich sequences of p85 and the SH3 domain of c-src. In vitro, the c-src SH3 domain stimulates PI3-K activity. Taken together, the current data suggest that c-src, via its SH3 domain, may participate in CSF-1-induced activation of PI3-K and that PI3-K and c-src are in the signaling pathway that subserves CSF-1-induced cytoskeletal changes in osteoclasts. PMID- 10830301 TI - Thyroid hormone regulation of phospholamban phosphorylation in the rat heart. AB - Thyroid hormone exerts predictable effects on the contractile performance of the heart in part by regulating the transcription of genes encoding specific calcium transporter proteins. In a rat model of hypothyroidism, left ventricular (LV) contractile function as measured by ejection fraction was decreased by 22% (P < 0.05), and this was returned to control values with T3 treatment. In confirmation of prior studies, LV phospholamban (PLB) protein content was significantly decreased by 25% and 40% compared with hypothyroid LV when the animals were treated with T3 at two doses, 2.5 and 7.0 microg/day, respectively. The ratio of sarcoplasmic reticulum calcium adenosine triphosphatase (SERCA2) to PLB protein content was thus increased by 171% and 207%, respectively (P < 0.01). Resolution of the phosphorylated PLB pentamers by SDS-PAGE showed that T3 infusion at 2.5 and 7.0 microg/day decreased (P < 0.001) the amount nonphosphorylated pentamers by 82% and 95%, respectively, in a dose-dependent manner. T3 treatment produced an increase in the proportion of highly phosphorylated PLB pentamers (more than five phosphates) when expressed as a fraction of total pentameric molecules (P < 0.05). Site-specific antibodies showed that the T3-induced increase in phosphorylated PLB pentamers was the result of an increase in both serine 16 and threonine 17 phosphorylation. We conclude that thyroid hormone, in addition to regulating the expression of cardiac PLB, is able to alter the degree of PLB phosphorylation, which correlates with enhancement of LV contractile function. These studies suggest that T3 may augment myocyte calcium cycling via changes in both cAMP- and calcium/calmodulin-dependent protein kinase activities. PMID- 10830302 TI - Nongenomic action of progesterone: activation of Xenopus oocyte phospholipase C through a plasma membrane-associated tyrosine kinase. AB - Using a plasma membrane-cortex preparation (wherein the nucleus and >90% of the total cell protein are removed), progesterone stimulated tyrosine kinase activity that stimulated phospholipase C. Although it has been known for over 20 yr that progesterone acts at the plasma membrane of Xenopus oocytes to induce oocyte maturation, this is the first report that progesterone stimulates this tyrosine kinase activity that is associated with the oocyte plasma membrane and cortex. A tyrosine kinase inhibitor (tyrphostin B46) inhibited steroid stimulation of tyrosine kinase and phospholipase C (PLC) activities, but did not block lipase C stimulation by G protein activators. A fusion protein that contains tandem N- and C-terminal SH2 domains of PLCgamma also blocked progesterone stimulation of PLC (a fusion protein with the SH2 domain from Shc was ineffective). Lowering the Ca2+ concentration in the medium inhibited progesterone, but not guanosine 5'-O (3-thiotriphosphate), stimulation of PLC, and the effects of progesterone and a G protein agonist were additive. However, neither progesterone nor insulin increased phosphotyrosine on PLCgamma. To evaluate another tyrosine kinase path involving phosphatidylinositol 3-kinase, we added wortmannin to our membrane preparation, but wortmannin did not inhibit progesterone's ability to activate PLC. PMID- 10830303 TI - Infusion of N-proopiomelanocortin-(1-77) increases adrenal weight and messenger ribonucleic acid levels of cytochrome P450 17alpha-hydroxylase in the sheep fetus during late gestation. AB - In the sheep there is a rapid increase in fetal adrenal growth and steroidogenesis during the last 10-15 days gestation (term = 147+/-3 days gestation). In the rat, peptides derived from the N-terminal region of POMC play a role in compensatory adrenal growth and in potentiation of ACTH-induced steroidogenesis. We therefore investigated the effects of infusion of bovine N POMC-(1-77) and its biosynthetic derivative, N-POMC-(1-49) on adrenal growth and on the expression of adrenal steroidogenic enzymes in the late gestation sheep fetus. Twenty-seven pregnant ewes were used in this study. Fetal vascular catheters were inserted between 116-125 days gestation, and purified bovine N POMC-(1-77) (2 microg/ml x h), N-POMC-(1-49) (2 microg/ml x h) and saline were each infused for 48 h between 136 and 138 days gestation. Intrafetal infusion of N-POMC-(1-77) resulted in an increased adrenal/fetal body weight ratio (94.6+/ 5.7 mg/kg) compared with that in saline-infused (75.6+/-1.8 mg/kg), but not N POMC-(1-49)-infused (82.7+/-6.1 mg/kg), fetal sheep. The ratio of CYP17 messenger RNA (mRNA) to 18S ribosomal RNA was also significantly higher in fetal adrenals ofthe N-POMC-(1-77)-infused group (49.1+/-4.7) compared with that in either the N POMC-(1-49)-infused (20.4+/-6.4) or saline-infused (15.2+/-4.4) group. There was no difference, however, in the ratios of adrenal CYP11A1 mRNA/3beta hydroxysteroid dehydrogenase/delta5,delta4-isomerase mRNA and CYP21A1 mRNA/18S ribosomal RNA among the N-POMC-(1-77)-, N-POMC-(1-49)-, and saline-infused groups. There was also no significant change in either plasma cortisol or ACTH concentrations in response to the infusion of either N-POMC-(1-77) or N-POMC-(1 49). In summary, intrafetal infusion of N-POMC-(1-77) stimulated fetal adrenal growth and resulted in a specific increase in adrenal CYP17 gene expression in late gestation. N-POMC-(1-77) may therefore play a modulatory role in the increase in fetal adrenal growth and steroidogenesis that occurs before birth. PMID- 10830304 TI - Npt2 gene disruption confers resistance to the inhibitory action of parathyroid hormone on renal sodium-phosphate cotransport. AB - PTH inhibition of renal sodium-phosphate (Na-Pi) cotransport is associated with the endocytic retrieval of the type II Na-Pi cotransporter, Npt2, from the renal brush border membrane into the late endosomal/lysosomal compartment. The aim of the present study was to determine whether mice homozygous for the disrupted Npt2 gene (Npt2-/-) exhibit decreased renal Pi reabsorption in response to PTH. We demonstrate that PTH has no effect on the serum Pi concentration, fractional excretion of Pi, or Na-dependent Pi transport in renal brush border membrane vesicles in Npt2-/- mice. In contrast, PTH elicits a fall in the serum Pi concentration, an increase in urinary Pi excretion, a decrease in brush border membrane Na-Pi cotransport, and a corresponding reduction in the relative abundance of Npt2 protein in wild-type mice (Npt2+/+). Both Npt2-/- and Npt2+/+ mice exhibit a significant rise in the urinary cAMP/creatinine ratio in response to PTH, indicating that generalized resistance to PTH cannot account for the absence of the PTH response in Npt2-/- mice. In addition, we demonstrate that Pi depleted normal mice respond to PTH with a decrease in renal brush border membrane Na-Pi cotransport and Npt2 protein, indicating that Pi deficiency per se does not account for PTH resistance in Npt2-/- mice. Taken together, our data provide compelling evidence that Npt2 gene expression is crucial for PTH effects on renal Pi handling. PMID- 10830305 TI - Down-regulation of thyroid transcription factor-1 gene expression in fetal lung hypoplasia is restored by glucocorticoids. AB - The thyroid transcription factor (TTF)-1 has an essential role in lung morphogenesis and development. It is involved in the transcription of surfactant proteins (SP), which are critical in respiratory function. Neonates with congenital diaphragmatic hernia die of respiratory failure caused by pulmonary hypoplasia with associated biochemical immaturity. To gain new insights into the causes of this disorder and the effect of prenatal hormonal treatment on reducing mortality in these infants, we evaluated the expression of TTF-1 as marker of lung morphogenesis and SP-B as marker of lung maturity. Using a rat model of lung immaturity, we show that TTF-1 and SP-B messenger RNA (mRNA) levels are drastically reduced in congenital lung hypoplasia. Interestingly, prenatal dexamethasone (Dex) treatment increased both TTF-1 and SP-B mRNAs over control levels when administered to rats with lung hypoplasia, but it had no effect on TTF-1 or a moderate effect on SP-B mRNA when administered to control rats. TRH alone also increases TTF-1 and SP-B mRNA levels but to a lesser extent than Dex. When administered together with Dex, TRH counteracts the induction observed with the glucocorticoid. The decrease in TTF-1 mRNA levels in lung hypoplasia is paralleled by a down-regulation of TTF-1 protein levels, as well as by a decrease in the TTF-1/DNA complex when the TTF-1-binding site of the SP-B promoter was used as a probe. Both parameters were reestablished after glucocorticoid treatment. Moreover, the regulation of TTF-1 gene expression described in this report is accompanied by the same regulation in its promoter activity, as demonstrated in transfection experiments performed in H-441 human lung-derived adenocarcinoma cells. In conclusion, our data demonstrate, for the first time, that lung hypoplasia and the associated respiratory dysfunction caused by SP-B deficiency are caused, in part, by down-regulation of TTF-1 gene expression. The observations that prenatal glucocorticoid treatment induces the expression of TTF 1 supports routine in utero glucocorticoid treatment of patients expected to have lung hypoplasia. PMID- 10830306 TI - Differential expression and regulation of estrogen receptors (ERs) in rat pituitary and cell lines: estrogen decreases ERalpha protein and estrogen responsiveness. AB - Estrogen (E) regulates the synthesis and secretion of several pituitary hormones during the reproductive cycle in a cell- and promoter-specific manner. One mechanism underlying cell specificity is the differential expression of estrogen receptor (ER) isoforms. We used in vivo and in vitro rodent pituitary cell models to examine the expression and regulation of ERalpha, ERbeta, and the pituitary specific ERalpha isoform, truncated estrogen receptor product-1 (TERP-1). In cycling female rat pituitaries, ERbeta messenger RNA (mRNA) levels fell 40% on the morning of proestrus and were suppressed by E or dihydrotestosterone in ovariectomized females. In lactotrope and gonadotrope cell lines (GH3, RC4B, LbetaT2), progesterone (P) or P plus E also suppressed ERbeta. TERP-1 mRNA increased 3-fold at proestrus and in response to E treatment in vivo and in cell lines. ERalpha mRNA levels were not regulated significantly by any treatment in vivo or in cell lines. However, E suppressed ERalpha protein levels in vivo and in cell lines, and reduction of ERalpha protein levels by E or the antiestrogen ICI182,780 reduced E-stimulated transcriptional activation of the PRL promoter in GH3 cells. TERP-1 and ERbeta protein levels were low to undetectable in cell lines, but E stimulated TERP-1. Because E treatment decreases ERbeta mRNA and ERalpha protein and increases levels of TERP-1 (which can suppress ERalpha/beta activity), the dynamic steroid-induced changes in ER expression in the rat pituitary during the midcycle gondaotropin/PRL surge may provide a means for ovarian steroids to limit positive feedback. PMID- 10830307 TI - Basic fibroblast growth factor stimulates collagenase-3 promoter activity in osteoblasts through an activator protein-1-binding site. AB - Basic fibroblast growth factor (bFGF) stimulates collagenase-3 synthesis in fetal rat osteoblast-enriched (Ob) cells. In this study we examined the mechanism of collagenase-3 regulation in Ob cells. bFGF at 0.6 nM or more increased the transcriptional rate of collagenase-3 by 3- to 7-fold. bFGF at 0.6 nM increased the activity of collagenase-3 promoter-luciferase reporter deletion constructs from -721 to -53 nucleotides transiently transfected into Ob cells by 3- to 5 fold. The minimal bFGF response was retained within the -53 to +28 sequence. Mutational analysis revealed that the bFGF effect was mediated through an activator protein-1 (AP-1)-binding site located at -48 to -42 nucleotides in the promoter. bFGF stimulated the binding of nuclear factors to the collagenase AP-1 site by 3- to 4-fold, as determined by electrophoretic mobility shift assays. Supershift analysis of nuclear extracts revealed that bFGF stimulates the occupancy of AP-1 site by c-Jun, JunB, JunD, c-Fos, FosB, and Fra2. In conclusion, bFGF increases collagenase-3 gene transcription, an effect mediated through an AP-1 site, due to the induction or activation of Jun and Fos family transcription factors. The stimulation of collagenase-3 synthesis by bFGF may be critical in mediating the actions of this growth factor in bone remodeling. PMID- 10830308 TI - Differential expression and regional distribution of steroid receptor coactivators SRC-1 and SRC-2 in brain and pituitary. AB - Members of the p160 family of steroid receptor coactivator proteins mediate the stimulatory effects on gene transcription brought about by nuclear receptors, which comprise all steroid receptors. Using in situ hybridization we have examined the neuroanatomical distribution of the messenger RNAs (mRNAs) for two functionally distinct splice variants of Steroid Receptor Coactivator 1 (SRC 1/NCoA-1) and of Steroid Receptor Coactivator 2 (SRC-2/NCoA-2/GRIP-1/TIF-2). Transcripts encoding these coactivators show highly differential expression patterns. SRC-2 mRNA is expressed at very low levels in brain, but shows expression in the anterior pituitary. SRC-la and le mRNA are expressed in many brain areas, including hippocampus, amygdala, hypothalamus, basal ganglia, and isocortex. Striking differences between SRC-1a and le expression were observed in several brain nuclei. Relative levels of SRC-1a mRNA were much higher in anterior pituitary, and the arcuate, paraventricular and ventromedial nucleus of the hypothalamus, the locus coeruleus and the trigeminal motor nucleus, all important targets of steroid hormones in the brain. SRC-le mRNA showed modest elevation of relative expression in the caudal nucleus accumbens (shell), basolateral amygdala, and some thalamic nuclei. The differential and uneven neuroanatomical distribution of these coactivators may underlie diversity and cell-specificity of steroid receptor mediated signals in the brain. PMID- 10830309 TI - The activation of phospholipase D by endothelin-1, angiotensin II, and platelet derived growth factor in vascular smooth muscle A10 cells is mediated by small G proteins of the ADP-ribosylation factor family. AB - We show here that A10 cells express the phospholipase D (PLD) isoforms PLD1b and PLD2. The activation of PLD in these cells by angiotensin II (AngII), endothelin 1 (ET-1), and platelet-derived growth factor (PDGF) was found to be sensitive to inhibitors of the activation of ADP-ribosylation factor (ARF) but not to blockers of Rho protein function. PDGF, AngII, and ET-1 induced the binding of ARF proteins to cell membranes in a permeabilized cell assay. Cells permeabilized and depleted of ARF were no longer sensitive to stimulation with AngII, ET-1, or PDGF, but the addition of recombinant myristoylated human ARF1 restored agonist dependent PLD activity. Expression of dominant negative ARF mutants blocked receptor-dependent activation of PLD. PLD activity was also potently stimulated by treatment with phorbol esters, but this activity was only partially inhibited by brefeldin A or by the overexpression of ARF dominant negative mutants. Transient expression of catalytically inactive mutants of PLD2, but not PLD1, inhibited significantly PDGF- and AngII-dependent PLD activity. We conclude: 1) the activation of PLD by cell surface receptors occurs primarily by an ARF dependent mechanism in A10 cells, whereas the activation of PLD by protein kinase C-dependent pathways is only partially dependent on the regulation of ARF proteins; and 2) cell surface receptors, such as AngII and PDGF, signal primarily via PLD2 in A10 cells. PMID- 10830310 TI - Annexin 1 (lipocortin 1) mediates the glucocorticoid inhibition of cyclic adenosine 3',5'-monophosphate-stimulated prolactin secretion. AB - Our previous studies have identified a role for annexin 1 (also called lipocortin 1) in the regulatory actions of glucocorticoids (GCs) on the release of PRL from the rat anterior pituitary gland. In the present study we used antisense and immunoneutralization strategies to extend this work. Exposure of rat anterior pituitary tissue to corticosterone (1 nM) or dexamethasone (100 nM) in vitro induced 1) de novo annexin 1 synthesis and 2) translocation of the protein from intracellular to pericellular sites. Both responses were prevented by the inclusion in the medium of an annexin 1 antisense oligodeoxynucleotide (ODN; 50 nM), but not by the corresponding sense and scrambled ODN sequences. Unlike the GCs, 17beta-estradiol, testosterone, and aldosterone (1 nM) had no effect on either the synthesis or the cellular disposition of annexin 1; moreover, none of the steroids or ODNs tested influenced the expression of annexin 5, a protein closely related to annexin 1. The increases in PRL release induced in vitro by drugs that signal via cAMP/protein kinase A [vasoactive intestinal polypeptide (10 nM), forskolin (100 microM), 8-bromo-cAMP (0.1 microM)] or phospholipase C (TRH, 10 nM) were attenuated by preincubation of the pituitary tissue with either corticosterone (1 nM) or dexamethasone (100 nM). The inhibitory actions of the steroids on the secretory responses to vasoactive intestinal polypeptide, forskolin, and 8-bromo-cAMP were specifically quenched by inclusion in the medium of the annexin 1 antisense ODN (50 nM) or a neutralizing antiannexin 1 monoclonal antibody (antiannexin 1 mAb, diluted 1:15,000). By contrast, the ability of the GCs to suppress the TRH-induced increase in PRL release was unaffected by both the annexin 1 antisense ODN and the antiannexin 1 mAb. In vivo, interleukin-1beta (10 ng, intracerebroventricularly) produced a significant increase in the serum PRL concentration (P < 0.01), which was prevented by pretreatment of the rats with corticosterone (100 microg/100 g BW, sc). The inhibitory actions of the steroid were specifically abrogated by peripheral administration of an antiannexin 1 antiserum (200 microl, sc); by contrast, when the antiserum was given centrally (3 microl, intracerebroventricularly), it was without effect. These results support our premise that annexin contributes to the regulatory actions of GCs on PRL secretion and suggest that it acts at point distal to the formation of cAMP. PMID- 10830311 TI - Luteinizing hormone (LH) drives diverse intracellular calcium second messenger signals in isolated porcine ovarian thecal cells: preferential recruitment of intracellular Ca2+ oscillatory cells by higher concentrations of LH. AB - The present study examines Ca2+ second messenger signaling driven by LH in isolated porcine thecal cells. To this end, we implemented semiquantitative fluorescent (fura-2) videomicroscopic imaging of single thecal cells in vitro. Stimulation of 388 cells with LH (5 microg/ml) elicited an intracellular Ca2+ ([Ca2+]i) signal in 85+/-5.3% of individual thecal cells (n = 11 experiments). Among 337 LH-responsive cells, we identified four predominant temporal modes of [Ca2+]i signaling: 1) [Ca2+]i oscillations with periodicities of 0.5 to 4.5 min( 1) (63+/-4.5%), 2) a [Ca2+]i spike followed by a sustained plateau (17+/-2.6%), 3) a [Ca2+]i spike only (5.8+/-2.6%); and 4) a [Ca2+]i plateau only (3.8+/-1.5%). The prevalence, but not the amplitude or frequency, of LH-induced [Ca2+]i oscillations in thecal cells was dependent on the agonist concentration. Reduced availability of extracellular Ca2+ induced by treatment with EGTA or cobaltous chloride did not block the initiation, but reversibly abolished ongoing [Ca2+]i oscillations (72% of cells) or increased the mean [Ca2+]i interspike periodicity from 1.09+/-0.16 to 0.59+/-0.07 min(-1) (P < 0.05). Putative phospholipase C inhibition with U-73122 (10 microM) also abolished or frequency-damped LH-driven [Ca2+]i oscillations in 95+/-4.7% of cells. [Ca2+]i oscillations in thecal cells were not abrogated by overnight pretreatment with pertussis toxin. We conclude that 1) thecal cells (unlike earlier findings in granulosa cells) manifest a diverse array of [Ca2+]i signaling responses to LH at the single cell level; 2) LH can dose dependently recruit an increasing number of individually [Ca2+]i oscillating thecal cells; 3) extracellular Ca2+ is required for LH to sustain (but not initiate) frequent and high amplitude [Ca2+] oscillations in thecal cells; and 4) these signaling actions of LH are mediated via phospholipase C, but not a pertussis-toxin sensitive mechanism. Accordingly, the present data extend the apparent complexity of LH-induced [Ca2+]i second messenger signaling and identify at the single cell level LH's dose-responsive drive of [Ca2+]i oscillations in gonadal cells. PMID- 10830312 TI - Impairment of cardiac function and bioenergetics in adult transgenic mice overexpressing the bovine growth hormone gene. AB - Cardiovascular abnormalities represent the major cause of death in patients with acromegaly. We evaluated cardiac structure, function, and energy status in adult transgenic mice overexpressing bovine GH (bGH) gene. Female transgenic mice expressing bGH gene (n = 11) 8 months old and aged matched controls (n = 11) were used. They were studied with two-dimensional guided M-mode and Doppler echocardiography. The animals (n = 6) for each group were examined with 31P magnetic resonance spectroscopy to determine the cardiac energy status. Transgenic mice had a significantly higher body weight (BW), 53.2+/-2.4 vs. 34.6+/-3.7 g (P < 0.0001) and hypertrophy of left ventricle (LV) compared with normal controls: LV mass/BW 5.6+/-1.6 vs. 2.7+/-0.2 mg/g, P < 0.01. Several indexes of systolic function were depressed in transgenic animals compared with controls mice such as shortening fraction 25+/-3.0% vs. 39.9+/-3.1%; ejection fraction, 57+/-9 vs. 77+/-5; mean velocity of circumferential shortening, 4.5+/ 0.8 vs. 7.0+/-1.1 circ/sec, p < 0.01. Creatine phosphate-to-ATP ratio was significantly lower in bGH overexpressing mice (1.3+/-0.08 vs. 2.1+/-0.23 in controls, P < 0.05). Ultrastructural examination of the hearts from transgenic mice revealed substantial changes of mitochondria. This study provides new insight into possible mechanisms behind the deteriorating effects of long exposure to high level of GH on heart function. PMID- 10830313 TI - Transforming growth factor-beta stimulates inorganic phosphate transport and expression of the type III phosphate transporter Glvr-1 in chondrogenic ATDC5 cells. AB - Members of the transforming growth factor (TGF)-beta family are important regulators of skeletal development. In this study, we investigated the effect of TGF-beta1 on inorganic phosphate (Pi) transport and on expression of the type III Pi carriers Glvr-1 and Ram-1 in murine ATDC5 chondrocytes. TGF-beta1 induced a selective, dose- and time-dependent increase in sodium-dependent Pi transport in ATDC5 cells. This response was dependent on RNA and protein synthesis and reflected a change in the maximal rate of the transport system, suggesting that TGF-beta1 induces the synthesis of new Pi carriers and their insertion into the plasma membrane. Consistently, Northern blotting analysis showed a dose-dependent increase in Glvr-1 messenger RNA expression in response to TGF-beta1, which preceded the maximal stimulation of Pi transport by several hours. Glvr-1 thus likely mediates at least part of the increase in Pi uptake induced by TGF-beta1. Ram-1 messenger RNA expression was not affected by TGF-beta1. TGF-beta1 activated the Smad signaling pathway and the mitogen-activated protein kinases ERK and p38 in ATDC5 cells. Unlike the regulation of Pi transport by receptor tyrosine kinase agonists in osteoblasts, the effect of TGF-beta1 on Pi uptake in ATDC5 cells did not involve protein kinase C or mitogen-activated protein kinases, suggesting that a specific, possibly Smad-dependent, signal mediates this response. In conclusion, TGF-beta1 stimulates Pi transport and Glvr-1 expression in chondrocytes, suggesting that, like proliferation, differentiation, and matrix synthesis, Pi handling is subject to regulation by TGF-beta3 family members in bone-forming cells. PMID- 10830314 TI - Nitric oxide and carbon monoxide have a stimulatory role in the hypothalamic pituitary-adrenal response to physico-emotional stressors in rats. AB - We tested the hypothesis that nitric oxide and carbon monoxide, which are produced in the brain by nitric oxide synthase (NOS) and heme oxygenase (HO), modulate the hypothalamic-pituitary-adrenal response to physico-emotional stressors by acting at the hypothalamus. Accordingly, we determined 1) whether the intracerebroventricular (icv) injection of NOS or HO inhibitors at doses that were confined to the brain attenuated electroshock-induced ACTH release; and 2) whether the decreases in this ACTH response were concurrent with decreases in NOS or HO activity levels at the hypothalamus. Icv injection of the NOS inhibitor Nomega-nitro-L-arginine-methylester (L-NAME; 50 microg) or the HO inhibitor tin protoporphyrin (SnPP; 20-25 microg) significantly blunted the plasma ACTH response to a 45-min session of intermittent electroshocks. Importantly, in these same animals there were concurrent decreases in hypothalamic NOS or HO activities, respectively. There were little or no effects of these inhibitors on anterior pituitary NOS or HO activities, indicating that there was only minimal leakage of the drug from the brain after icv administration. The specificity of action of these inhibitors was confirmed by the fact that SnPP did not affect NOS activity, and L-NAME did not affect HO activity. Finally, L-NAME produced no effect, whereas SnPP produced only transient increases in blood pressure, suggesting that these inhibitors do not affect activity indirectly through alterations in blood pressure. These data support the hypothesis that in the whole animal, both NO and CO exert a stimulatory influence on the acute ACTH response to physico-emotional stressors, and that the hypothalamus is the critical site of their actions. PMID- 10830315 TI - CCN proteins are distinct from and should not be considered members of the insulin-like growth factor-binding protein superfamily. PMID- 10830316 TI - Overexpression of insulin-like growth factor binding protein-5 helps accelerate progression to androgen-independence in the human prostate LNCaP tumor model through activation of phosphatidylinositol 3'-kinase pathway. AB - Although insulin-like growth factor (IGF) binding protein-5 (IGFBP-5) is highly up-regulated in normal and malignant prostate tissues after androgen withdrawal, its functional role in castration-induced apoptosis and androgen-independent progression remains undefined. To analyze the functional significance of IGFBP-5 overexpression in IGF-I-mediated mitogenesis and progression to androgen independence, IGFBP-5-overexpressing human androgen-dependent LNCaP prostate cancer cells were generated by stable transfection. The growth rates of IGFBP-5 transfected LNCaP cells were significantly faster, compared with either the parental or vector-only transfected LNCaP cells in both the presence and absence ofdihydrotestosterone. IGFBP-5-induced increases in LNCaP cell proliferation occurs through both IGF-I-dependent and -independent pathways, with corresponding increases in the cyclin D1 messenger RNA expression and the fraction of cells in S + G2/M phases of the cell cycle. Changes in Akt/protein kinase B, a downstream component of phosphatidylinositol 3'-kinase (PI3K) pathway, in the LNCaP sublines also paralleled changes in their growth rates. Although treatment with a PI3K inhibitor induced apoptosis in both control and IGFBP-5-overexpressing LNCaP cells, this PI3K inhibitor-induced apoptosis was prevented by exogenous IGF-I treatment only in IGFBP-5 transfectants, suggesting that IGFBP-5 overexpression can potentiate the antiapoptotic effects of IGF-I. Furthermore, tumor growth and serum prostate-specific antigen levels increased several fold faster in mice bearing IGFBP-5-transfected LNCaP tumors after castration, despite having similar tumor incidence and tumor growth rates with controls when grown in intact mice before castration. Collectively, these data suggest that IGFBP-5 overexpression in prostate cancer cells after castration is an adaptive cell survival mechanism that helps potentiate the antiapoptotic and mitogenic effects of IGF-I, thereby accelerating progression to androgen independence through activation of the PI3K Akt/ protein kinase B signaling pathway. PMID- 10830317 TI - Identification of an estrogen-mediated deoxyribonucleic acid-binding independent transactivation pathway on the epidermal growth factor receptor gene promoter. AB - To investigate the estrogenic effects on the transcriptional regulation of the epidermal growth factor (EGF) receptor (EGFR) gene, we assayed its promoter ability to direct transcription of the luciferase reporter gene after transfection into HeLa cells. Our studies demonstrated a dose-dependent activation of the EGFR gene transcription by ligand-bound estrogen receptor alpha (ERalpha). This action was retained by the 36-bp core promoter fragment and did not require the receptor DNA binding domain, as demonstrated by analyzing the role of ERalpha deletion mutants on EGFR gene promoter-derived constructs. The 36 bp promoter fragment does not contain an estrogen response element but an imperfect thyroid hormone response element half-site that overlaps the Sp1 binding site. ERalpha does not bind this imperfect thyroid hormone response element half-site but is able to enhance binding of Sp1 to its site, in gel mobility shift assays, suggesting that the mechanism by which the receptor stimulated the transcription involved protein-protein interactions that replaced DNA binding. To explain this action, we propose a model in which induction of the EGFR gene expression by estrogens in HeLa cells is dependent upon the formation of a transcriptionally active ERalpha-Sp1 complex that binds to the GC-rich (Sp1) region of the minimal promoter. PMID- 10830318 TI - Augmentation of thyroid hormone receptor-mediated transcription by Ca2+/calmodulin-dependent protein kinase type IV. AB - Thyroid hormone receptor (TR), a ligand-mediated transcription factor, binds to a DNA sequence known as a thyroid-hormone response element (TRE) to activate or repress transcription of target genes. Recently, studies have shown that Ca2+/calmodulin-dependent protein kinases (CaMKs) may be involved in regulating gene transcription via phosphorylation of specific transcription factors, including RORalpha, a retinoic acid-related orphan nuclear hormone receptor. In this light, we examined the effect of CaMK type IV (CaMKIV) and RORalpha, which also shown to influence thyroid hormone action, on TR-mediated transcription using a transient transfection assay. Expression vectors containing TR, vitamin D receptor (VDR), and estrogen receptor (ER) were cotransfected in CV-1 cells with RORalpha and/or constitutively active CaMKIV and thymidine kinase promotor luciferase reporter vector containing their cognate response elements. When CaMKIV or RORalpha was co-transfected with TR, the T3-induced transcription was significantly augmented compared to that induced by TR alone. When both were co transfected with TR, T3-induced transcription was augmented additively. In contrast, the augmentation by CaMKIV or ROR on ligand-induced transcription was not detected with VDR and ER. Hence, these results indicate that the augmentation mediated by CaMKIV and RORalpha is specific for TR-mediated transcription on TRE. Our results suggest that CaMKIV, as well as RORalpha, play important roles in TR mediated transcription on TREs. PMID- 10830319 TI - Localizing and lateralizing value of epileptic symptoms in temporal lobe epilepsy. AB - The symptoms and signs associated with all stages of a temporal lobe seizure may be helpful in determining both the localization and lateralization of seizure onset. Auras, when present, may be very suggestive of temporal lobe onset and may further localize to a mesiobasal or lateral temporal lobe site of onset. During the ictus, automatisms and motor phenomena may be highly indicative of temporal lobe seizure activity and may even help lateralize the discharge. In the post ictal period, motor paresis and aphasia are helpful in lateralization. Video E.E.G. data has provided extensive information on the utility of ictal symptomatology in seizure localization. Thus, the seizure semiology provides important adjunctive information in evaluating patients for epilepsy surgery and should be concordant with information obtained from ictal EEG, neuroimaging and neuropsychology. PMID- 10830320 TI - Epidemiology of temporal lobe epilepsy. AB - To fulfill its task of informed clinical decision making and resource allocation, epidemiological studies in epilepsy must adhere to a series of methodological standards. These are reviewed. Because seizure and epilepsy classification systems may be viewed as extensions of the diagnosis, they have direct implications in the acquisition and interpretation of epidemiologic data. The International League Against Epilepsy (ILAE) classification systems are analyzed in this light. Finally, the lack of Canadian epidemiological studies is addressed, and the relevance and potential of Canadian epidemiological data in epilepsy is discussed. PMID- 10830321 TI - Temporal lobe epilepsy surgery: definition of candidacy. AB - Medical intractability is one of the absolute indications for considering temporal lobe epilepsy surgery. This is a relative concept that has to be highly individualized. It is quite easy to determine when a patient's seizures are fully controlled. On the other hand, "continuing seizures are not necessarily a measure of intractability or disability". A positive decision to operate would be based on some of the following factors: assurance of a firm diagnosis, seizures that are frequent and disabling, and seizures occurring in patients who are drug refractory to optimal anti-epileptic medications and dosages. PMID- 10830322 TI - Pediatric candidates for temporal lobe epilepsy surgery. AB - Temporal lobectomy is an under-valued and effective therapy for selected children with temporal lobe seizures. This article details three components of the identification and evaluation of such children. The ictal symptoms and signs remains an essential component. This includes an aura suggesting limbic system involvement, and the nature of automatisms and other motor phenomena which may have lateralising value. Electroencephalography continues to accurately identify a principally epileptogenic focus in children and adults. The value of high quality magnetic resonance imaging is emphasized. Invasive studies are required in only a minority of cases. Several studies have revealed that 73-100% of carefully selected children receive benefit from this procedure whereas significant neurosurgical complications rarely occur. PMID- 10830323 TI - Scalp EEG in temporal lobe epilepsy surgery. AB - Electroencephalography (EEG) with standard scalp and additional noninvasive electrodes plays a major role in the selection of patients for temporal lobe epilepsy surgery. Recent studies have provided data supporting the value of interictal and postictal EEG in assessing the site of ictal onset. Scalp ictal rhythms are morphologically complex but at least one pattern (a five cycles/second rhythm maximum at the sphenoidal or anterior temporal electrode) occurs in >50% of patients and has a high predictive value and interobserver reliability for temporal lobe originating seizures. Thorough interictal and ictal scalp EEG evaluation, in conjunction with modern neuroimaging, is sufficient for proceeding to surgery without invasive recordings in some patients. Further studies are required to define the scalp ictal characteristics of mesial vs. lateral temporal lobe epilepsy. PMID- 10830324 TI - Invasive electrographic recording techniques in temporal lobe epilepsy. AB - The management of uncontrolled partial epilepsy is a process dependent on a multidisciplinary and analytic approach. It is necessary to understand which lesions are epileptogenic, and if they are indeed responsible for the generation of seizures. In addition to localizing seizure onset, the functional and eloquent areas of the brain need to be identified. As in many other centres, we perform resective surgeries on the basis of combined information derived from seizure semiology, EEG abnormalities, neuroimaging and other tests of cerebral function. If surface EEG recording yields inconclusive or ambiguous results, then invasive intracranial techniques using intracerebral depth or subdural electrodes can be used to improve diagnostic or prognostic accuracy. The indications, principles, results and complications of these recording techniques based on extensive experience at two epilepsy surgery centres are reviewed. PMID- 10830325 TI - MRI techniques: bilateral findings and "normal findings". AB - Magnetic resonance imaging (MRI) techniques allow for significantly better imaging of the temporal lobe compared to computed tomography (CT) or other non invasive modalities. For detection of foreign tissue lesions, MRI surpasses CT. For the highest non-invasive yield for detection of mesial temporal sclerosis, optimal sequences that should be employed are a heavily T1-weighted volumetric acquisition (to enable both volumetric calculation of hippocampal volume, and, if needed, intracranial volume), T2-weighted coronal sequences, with or without T2 mapping, fluid-attenuated inversion recovery (FLAIR) and, to exclude subtle susceptibility effects from hematoma or cavernoma, gradient echo scans. Magnetic resonance spectroscopy (MRS) may show a decrease in N-acetyl aspartate (NAA) concentration, or NAA: Choline + creatine ratio. Functional MRI is a new and exciting tool that offers the promise of accurately localizing hemispheric functions; its role in the preoperative evaluation of temporal lobe seizures remains uncertain at present. PMID- 10830326 TI - Neuropsychological assessment for temporal lobe epilepsy surgery. AB - Neuropsychological assessment consists of a comprehensive evaluation of cognitive functioning and most often some evaluation of motor skills and sensory status also. Cognitive functions sampled typically include "intelligence" (IQ tests), attention, language skills, visuospatial abilities, "executive skills" and other abilities associated with frontal-lobe function, and learning and memory. Thus, the assessment samples vary widely among a variety of functions, providing a comprehensive picture of an individual's strengths and weaknesses. The resulting pattern points to the probable site of epileptic focus. Neuropsychological findings also serve to predict the risk for postsurgical cognitive decline and, when performance before and after operation is compared, they provide data on the impact of surgery upon cognitive functioning. Comprehensive evaluation of learning and memory is particularly important in this context, because of the frequency of temporal lobe epilepsy and the prominence of memory dysfunction associated with it. In addition, patients slated for elective surgery may also undergo an intracarotid amobarbital procedure (IAP), which is performed to determine the side of cerebral dominance for language and to test the memory capabilities of each hemisphere alone. All of these specialized neuropsychological tools are discussed in this paper. PMID- 10830327 TI - Psychiatric assessment of candidates for epilepsy surgery. AB - Patients with medically intractable epilepsy often present with comorbid psychiatric diseases. When referred to a program for the surgical treatment of the epilepsies, these patients benefit from a pre-admission psychiatric assessment with a view to lessen the chances of a psychiatric crisis during the pre- and postoperative investigations. This article proposes a practical approach to the psychiatric assessment and monitoring of adult candidates to the surgery of epilepsy. It emphasizes, in agreement with a world literature review, that definitive psychiatric contraindications to this elective surgery are few, and that adverse long-term psychiatric outcomes are less frequent when good seizure outcome is achieved. PMID- 10830328 TI - Communication and mental preparation issues for the epilepsy surgery patient. AB - This paper reviews the importance of adequate communication between the treatment team and the patient and family in epilepsy surgery. Ensuring that patient and family adequately understand the diagnosis, natural history, management alternatives and the risks and outcomes of surgery is a challenge for each epilepsy team. Guidelines as to process and content from the authors' experience and the participants in the workshop are presented. PMID- 10830329 TI - Awake craniotomy: controversies, indications and techniques in the surgical treatment of temporal lobe epilepsy. AB - In 1886, Victor Horsley excised an epileptogenic posttraumatic cortical scar in a 23-year-old man under general anaesthesia and discussed his choice of anaesthesia: "I have not employed ether in operations on man, fearing that it would tend to cause cerebral excitement; chloroform, of course, producing on the contrary, well-marked depression." His concerns regarding anaesthesia are reiterated 100 years later as evidenced by the ongoing controversy over the choice of anaesthetic in surgical procedures for epilepsy. The current controversies regarding the necessity for local anaesthesia in temporal lobe epilepsy operations concern the utility of electrocorticography in surgical decision making, its relationship to seizure outcome and the value of intraoperative language mapping in dominant temporal lobe resections. The increasing sophistication of pre-operative investigation and localization of both areas of epileptogenesis and normal brain function and the introduction of minimally invasive surgical techniques and smaller focal resections are changing the indications for local anaesthesia in temporal lobe epilepsy. Thus, indications which were previously absolute are now perhaps relative. This article reviews the current indications for craniotomy under local anaesthesia in the surgical treatment of temporal lobe epilepsy. PMID- 10830330 TI - Anesthesia for temporal lobe epilepsy surgery. AB - Anesthetic considerations for temporal lobectomy for refractory epilepsy include ensuring a safe and comfortable perioperative experience for the patient, providing suitable operating conditions for the surgeon, avoiding interference with intraoperative electrocorticographic (ECoG) recordings and facilitating intraoperative functional cortical mapping, if performed. Providing the conditions that simultaneously meet these requirements, using general anesthesia or local anesthesia with sedation, remains a significant challenge for the neuroanesthetist. We review issues pertinent to the choice of anesthetic technique for these procedures. PMID- 10830331 TI - Transcortical selective amygdalohippocampectomy in temporal lobe epilepsy. AB - Numerous studies of the electrophysiology and neuropathology of temporal lobe epilepsy have demonstrated the mesial temporal structures to be the site of seizure origin in the majority of cases. This is the rationale for a transcortical selective approach, first introduced by Niemeyer, for removal of the hippocampus and amygdala. Series from a number of centers have demonstrated the efficacy of selective amygdalohippocampectomy compared to a more traditional resection. The technique described here and used at the Montreal Neurological Institute (MNI) utilizes a strictly endopial resection of the hippocampal formation and amygdala in addition to computer image guidance to perform the procedure. Ninety-five percent of patients at the MNI who underwent selective amygdalohippocampectomy realized a cessation of seizures, or greater than 90% reduction, with minimal risk of complications. PMID- 10830332 TI - Surgical pitfalls, their consequences, transient complications. AB - The methodology of this paper is based entirely on the experiential backgrounds of the authors. It outlines those factors which have become recognized as potentially important issues to patients who are considering recommendations of surgical treatment for their intractable epilepsy. Thus, on the one hand, it includes the important generic aspects of Informed Consent, while on the other hand there must be a very comprehensive and, when the operation is to be carried out under local anesthesia, a very detailed explanation of the preparation and the sequential steps in the surgical procedure. This should also entail a brief description of the roles of the various "team" members during the operative procedure. There are well-recognized complications associated with the various surgical procedures for the treatment of epilepsy. Further, there are predictable deficits following some of these procedures, some of which might be permanent and some of which may be transient. These pitfalls are briefly discussed. PMID- 10830333 TI - Stereotactic surgery for temporal lobe epilepsy. AB - In light of the recent resurgence of interest in stereotactic functional procedures, the authors have reviewed the role of stereotactic ablative surgery in the treatment of temporal lobe epilepsy. Literature pertaining to stereotactic amygdalotomy, hippocampotomy and fornicotomy is reviewed and summarized. However, the results presented in those early studies are difficult to interpret given the surgical techniques and outcome assessment used. Modern stereotactic ablative surgery using current image-guided technology offers the opportunity to revisit some of these techniques. In recent studies of stereotactic amygdalohippocampotomy, outcomes are not as favorable as those obtained with standard temporal resections. However, these relatively discrete ablative techniques may shed light on the anatomical substrate underlying temporal lobe epilepsy. PMID- 10830334 TI - Intraoperative electrocorticography in temporal lobe epilepsy surgery. AB - Although in clinical use for many years, the validity of intraoperative electrocorticography (ECoG) in guiding resective temporal lobe epilepsy (TLE) surgery is uncertain. Advances in neuroimaging and extraoperative intracranial recordings have contributed greatly to the identification of epileptogenic lesions and cortex, clarifying the limitations of a brief intraoperative interictal recording. Studies of undifferentiated ECoG findings (which classify all interictal cortical spike discharges as equal) tend to not support this method. This article reviews ECoG and presents data from 86 TLE surgeries at the University of British Columbia suggesting that differentiation of ECoG features may enhance the contribution of this time honored method. Specifically, independent foci may be more important for epileptogenesis than synchronous foci, and postexcision activation appears to be a benign phenomenon, while residual spikes unaltered by the resection correlate with a greater proportion of seizure recurrence. PMID- 10830335 TI - Seizure control as an indicator of therapeutic usefulness. AB - Seizure control, in addition to quality of life, is an important outcome after epilepsy surgery. However, seizure measurement is not straightforward. We explore some important difficulties in obtaining unbiased, reliable estimates of seizure frequency, and discuss aspects of data analysis as it pertains to validity and clinical meaningfulness. As seizure severity is an integral component of seizure outcome assessment, we discuss aspects of measurement of this outcome. Suggestions for improving validity are offered, based on the literature and on methodological common sense. Finally, we look at the Canadian perspective and the potential for adopting methodology that allows for data pooling and for more powerful analyses. PMID- 10830336 TI - Management of antiepileptic drugs following epilepsy surgery: a review. AB - The favourable impact of surgery for intractable epilepsy on seizures is well documented. However, few studies have determined what changes in antiepileptic drug (AED) therapy occur following surgery. Alterations in AED blood levels in the immediate postoperative period can result in breakthrough seizures. In long term follow-up, one-third to one-half of seizure-free patients after temporal lobectomy, the most common type of epilepsy surgery, still require AED treatment which usually has been reduced from polytherapy to monotherapy. In our study of 93 patients followed two years after temporal lobectomy, polytherapy decreased from 78% before surgery to 14% postoperatively, while medication was discontinued in 44%. Early reduction from polytherapy to monotherapy can often be carried out in the immediate postoperative period but the most appropriate timing of cessation of AED treatment has yet to be determined. Patients undergoing extratemporal resections and other forms of epilepsy surgery associated with less favourable postoperative seizure control may achieve comparatively less reduction in antiepileptic medication. We generally do not recommend stopping AEDs prior to one year after surgery. PMID- 10830337 TI - Measuring quality of life in epilepsy surgery patients. AB - Treatment of chronic conditions often entails trade-offs between benefits and risks or side effects. This is of particular importance in epilepsy. Consequently, assessment of effectiveness must incorporate this trade-off as perceived by the recipient of the intervention, ie., the patient. Measurement of health related quality of life (HRQOL) aims at capturing such patient-centred effects of therapy. Conceptual aspects of HRQOL are described and issues relevant to outcomes of epilepsy surgery are addressed. Finally, practical aspects of measurement in the context of epilepsy surgery are discussed. PMID- 10830338 TI - Psychological adjustment to success and to failure following epilepsy surgery. AB - Treatment-refractory epilepsy can directly impair psychological adjustment and quality of life. Epilepsy surgery is viewed by the patient as a stressful life event, arrived at after extensive deliberation. It is accompanied by expectations and anxiety for patients and their families. We suggest that the success of epilepsy surgery (as reflected by patient-perceived impact on quality of life) depends not only on reduced seizure frequency, but on personality factors and psychosocial issues. This paper reviews our research on these factors, along with suggestions for psychological assessment (of quality of life, and of personality) and psychological treatment. Studies of quality of life in patients undergoing temporal lobectomy are also examined. Finally, factors leading to the perception of "failed" epilepsy surgery are considered, along with suggested ways to manage such cases. PMID- 10830339 TI - Endarterectomy for asymptomatic carotid stenosis in the real world. PMID- 10830340 TI - Enhancing recovery after stroke with noradrenergic pharmacotherapy: a new frontier? AB - Despite much progress in stroke prevention and acute intervention, recovery and rehabilitation have traditionally received relatively little scientific attention. There is now increasing interest in the development of stroke recovery drugs and innovative rehabilitation techniques to promote functional recovery after completed stroke. Experimental work over the past two decades indicates that pharmacologic intervention to enhance recovery may be possible in the subacute stage, days to weeks poststroke, after irreversible injury has occurred. This paper discusses the concept of "rehabilitation pharmacology" and reviews the growing literature from animal studies and pilot clinical trials on noradrenergic pharmacotherapy, a new experimental strategy in stroke rehabilitation. Amphetamine, a monoamine agonist that increases brain norepinephrine levels, is the most extensively studied drug shown to promote recovery of function in animal models of focal brain injury. Further research is needed to investigate the mechanisms and clinical efficacy of amphetamine and other novel therapeutic interventions on the recovery process. PMID- 10830341 TI - Surgical treatment of epilepsy in pediatric patients. AB - Surgery has become an accepted treatment modality for carefully selected adults with intractable focal epilepsy. More recently, increasing numbers of pediatric patients with intractable epilepsy are also being referred for surgical consideration. Key elements of surgical candidacy include medically intractable focal epilepsy, a localized epileptogenic zone, and a low risk for new postoperative neurologic deficits. The most common etiologies of the epilepsies in pediatric surgical candidates are malformation of cortical development and low grade tumor but some patients with childhood onset temporal lobe epilepsy due to hippocampal sclerosis also present for early surgery. Based on results from several recent pediatric surgical series, the chance for favorable seizure outcome after surgery is not adversely affected by young age, with seizure-free postoperative outcome reported for 60% to 65% of infants, 59% to 67% of children, and 69% of adolescents, compared to 64% reported in a large, predominantly adult series. Some subgroups of patients have higher percentages of seizure-free outcome, including those with hippocampal sclerosis or low grade tumor. In addition to seizures, developmental issues are also a major concern in children with intractable epilepsy. Few quantitative data are available, but some anecdotal experience suggests that surgical relief of catastrophic epilepsy may result in resumption of developmental progression after surgery, although the rate of development often remains abnormal. In one series, best developmental outcomes were seen in patients with earliest surgery and highest level of preoperative development. For each patient, the timing of surgery must be carefully considered, based on a full assessment of the relative risks and benefits, derived from a detailed presurgical evaluation. PMID- 10830342 TI - Are the triptans for migraine therapy worth the cost? AB - The triptans represent a major advance in migraine therapy but their cost per dose greatly exceeds that of many older treatments. There is evidence that for a significant proportion of migraine patients these new drugs can show a positive cost benefit and also improve quality of life. Cost benefit would be expected to be greatest in patients with more severe migraine attacks. PMID- 10830343 TI - Attitudes of Canadian and U.S. neurologists regarding carotid endarterectomy for asymptomatic stenosis. AB - BACKGROUND: The American Heart Association carotid endarterectomy (CE) guidelines endorse CE for asymptomatic carotid stenosis if the procedure can be performed with low morbidity. However, the Canadian Stroke Consortium has published a consensus against CE for asymptomatic stenosis. The views of practicing neurologists in the two countries on this subject are unclear. METHODS: A survey was undertaken of 270 neurologists from either Florida or Indiana and 180 neurologists from either Ontario or Quebec. RESULTS: The survey was returned by 36% of neurologists. Both Florida (65%) and Indiana neurologists (35%) were significantly more likely than Canadian neurologists (11%) to sometimes/often refer patients for surgery(p<0.001). Neurologists from Florida relied more on noninvasive methods of carotid stenosis assessment (36%) than Canadian neurologists (12%, p=0.003), who preferred angiography. Neurologists from Florida more often cited medicolegal concerns as a reason for referring patients for surgery (27%), compared to Canadian neurologists (3%, p=0.0001). CONCLUSIONS: Practices pertaining to carotid stenosis evaluation and management differ both regionally and by country. Canadian neurologists refer fewer asymptomatic patients for CE and rely more on angiography as a preoperative diagnostic tool. The potential of medicolegal liability is a greater force in clinical decision making for certain U.S. neurologists, compared to their Canadian counterparts. These differences may partly explain the variations in CE utilization in the two countries. PMID- 10830344 TI - Diagnostic strategies in young patients with ischemic stroke in Canada. AB - BACKGROUND: A preliminary national survey of ischemic stroke in the young (15-45 years) undertaken by the Canadian Stroke Consortium indicated that in 44% of 356 patients, no cause was found. OBJECTIVE: To determine the reason for this high incidence of diagnostic uncertainty in young patients with ischemic stroke. METHODS: Neurologists in the ten Canadian stroke centers completed a detailed questionnaire for patients aged 15-45 years admitted to hospital between January 1993 and December 1997. Using a stepwise diagnostic algorithm incorporating clinical, neuroimaging, neurovascular and laboratory data, we divided patients into three groups: (1) those with established cause for the ischemic stroke, (2) those who remained unexplained despite adequate investigation, (3) those who remained unexplained but were, in our opinion, under-investigated. RESULTS: In 197 patients (56%), an identified cause was established including cardioembolic sources (14%), extracranial arterial dissection (13%), lacunar infarcts (8%) atherosclerosis (6%). A miscellaneous group of 15%, included cerebral venous thrombosis, coagulopathies, vasculitis and others. In 159 patients (44%) with no apparent cause for their stroke, we considered only 81 (23%) adequately investigated, and 78 (21%) inadequately investigated. CONCLUSION: About one in five young patients was inadequately investigated by a stroke-oriented group of neurologists. The major problem appears to be restriction of investigations to neuroimaging alone (usually computerized cerebral tomography), without further tests such as cerebral angiography and cardiac imaging. PMID- 10830345 TI - Percutaeous radiofrequency facet rhizotomy--experience with 118 procdedures and reappraisal of its value. AB - BACKGROUND: There have been many reports of percutaneous radiofrequency facet rhizotomy, perhaps better referred to as facet denervation, usually performed under general anaesthesia, with inconsistent success rates. OBJECTIVES: To report the authors' outcome data using both general and local anaesthesia and to reassess the value of this controversial procedure. METHODS: Our experience with 118 consecutive percutaneous radiofrequency facet rhizotomies performed on 90 patients in the Toronto Western Hospital was analyzed. Sixty percent of the procedures were performed under general anaesthesia, 40% under local anaesthesia. All patients had been temporarily virtually relieved of pain after local anaesthetic blockade of the subject facets by an independent radiologist. RESULTS: The patients were monitored from 1-33 (mean 5.6) months after surgery, with complete elimination or a greater than 50% subjective reduction of pain considered the criteria for success. For the first or only procedure this was 41% overall, 37% in cases done under local anaesthesia, 46% in cases done under general anaesthesia (difference not statistically significant p=0.52). There was no statistically significant difference in success rates for procedures performed in the cervical, thoracic or lumbosacral facets, with unilateral versus bilateral denervations, when two to three as compared with more than three facets were denervated, nor for operations done in patients who had had previous spinal surgery compared with those who had not. Results were not better regardless of whether hyperextension of the spine aggravated the patient's preoperative pain or not, and when the procedures were repeated in the same patient outcomes tended to be consistent, arguing against repetition of failed facet denervations. The morbidity was low, the chief problem being sensory loss and transient neuropathic pain in the distribution of cutaneous branches of posterior rami in the cervical and thoracic areas; mortality was zero. CONCLUSIONS: Percutaneous radiofrequency facet denervation is simple and safe, still worth considering in patients with disabling spinal pain that fails to respond to conservative treatment. The use of general anaesthesia shortens the operating time and the patient's discomfort without impairing success rate. PMID- 10830346 TI - Electroclinical analysis of postictal noserubbing. AB - BACKGROUND: Postictal noserubbing (PIN) has been identified as a good, albeit imperfect, lateralizing and localizing sign in human partial epilepsy, possibly related to ictal autonomic activation. METHODS: PIN was studied prospectively in a group of consecutive patients admitted for video-EEG monitoring, with the laterality of noserubbing correlated with electrographic sites of seizure onset, intra- and interhemispheric spread, and sites of seizure termination. RESULTS: PIN was significantly more frequent in temporal than extratemporal epilepsy (p<0.001; 23/41 (56%) patients and 41/197 (21%) seizures in temporal lobe epilepsy compared with 4/34 (12%) patients and 12/167 (7%) seizures in extratemporal epilepsy). The hand used to rub the nose was ipsilateral to the side of seizure onset in 83% of both temporal and extratemporal seizures. Seizures with contralateral PIN correlated with spread to the contralateral temporal lobe on scalp EEG (p<0.04). All extratemporal seizures with PIN showed spread to temporal lobe structures. One patient investigated with intracranial electrodes showed PIN only when ictal activity spread to involve the amygdala: seizures confined to the hippocampus were not associated with PIN. PIN was not observed in 63 nonepileptic events in 17 patients. Unexpectedly, one patient with primary generalized epilepsy showed typical PIN after 1/3 recorded absence seizures. CONCLUSIONS: This study confirms PIN as a good indicator of ipsilateral temporal lobe seizure onset. Instances of false lateralization and localization appear to reflect seizure spread to contralateral or ipsilateral temporal lobe structures, respectively. Involvement of the amygdala appears to be of prime importance for induction of PIN. PMID- 10830347 TI - Head tremor in cervical dystonia. AB - OBJECTIVE: To compare the clinical characteristics, natural history, and therapeutic outcome of patients with cervical dystonia (CD) with head tremor (HT+) and without head tremor (HT-). METHODS: We prospectively evaluated 114 consecutive patients of CD over a 9-month period with a detailed questionnaire. Chi-square and t-tests were employed for statistical analysis. RESULTS: Seventy eight (68.4%) patients had head tremor and 27 of them (34.6%) had tremor as one of the first symptoms. Age at onset of symptoms were similar in HT+ and HT- groups; however there was a higher prevalence in women in the former group (66.7% vs. 41.7%; p=0.01). HT+ patients had more frequent positive family history of essential-like hand/head tremor (21.8% vs. 5.5%; p<0.05), associated neck pain (92.3% vs. 77.8%: p<0.05), and essential-like hand tremor (40% vs. 8.3%; p<0.001). They also appeared to have more frequent history of preceding head/neck trauma (14.1% vs. 8.3%), frequent head rotation (88.5% vs. 69.4%) and antecollis (12.8% vs. 5.5%) but less often head tilt (37.2% vs. 47.2%) and gestes antagonistes (60.2% vs. 75%) than the HT- patients; however these differences were not statistically significant. The frequency of prior psychiatric illnesses, the incidence of dystonias in other parts of the body, frequency of retrocollis and shoulder elevation, and spontaneous remission were similar in the two groups. CONCLUSION: Head tremor is common in CD and is more commonly associated with hand tremor and family history of tremor or other movement disorders. This supports a possible genetic association between CD and essential tremor (ET). Linkage studies are required to evaluate the genetic association between CD and ET. PMID- 10830348 TI - Intracranial pressure monitoring in severe traumatic brain injury--results of a Canadian survey. AB - OBJECTIVE: The purpose of this study was to obtain information from Canadian neurosurgeons regarding their opinions on, and utilization of, intracranial pressure (ICP) monitoring for severe traumatic brain injury (TBI). METHODS: A brief survey was sent to practicing Canadian neurosurgeons questioning them about their utilization of, and confidence in, intracranial pressure monitoring in the management of patients with severe TBI. RESULTS: One hundred and ninety-six surveys were mailed. There were 103 responses for a response rate of 52.6%. The vast majority of responding neurosurgeons (98.1%) utilized ICP monitoring in the management of patients with severe TBI, with most (63.4%) using it in more than 75% of their patients, 14.9% using it in 50-75% of patients, 14.9% in 25-50% of patients, and 6.9% using it in less than 25% of patients. The level of confidence that routine monitoring improves outcome from severe TBI ranged from 23.3% having a low level of confidence, 56.3% having an intermediate level of confidence, to 20.4% having a high level of confidence. Most respondents (78.6%) felt that some form of prospective trial evaluating the role of ICP monitoring in improving outcome from severe TBI was warranted; 17.4% felt such a trial was not warranted and 3.9% were uncertain. CONCLUSIONS: While ICP monitoring has gained almost universal acceptance among responding Canadian neurosurgeons, their level of confidence that routine monitoring improves outcome from severe TBI was quite variable, with only 20.4% of respondents having a high level of confidence. Over 75% of respondents felt that some form of prospective trial evaluating the utility of ICP monitoring is warranted. This information is being used in consideration of a prospective trial addressing this issue. PMID- 10830349 TI - Cognitive function in Nigerians with newly diagnosed epilepsy. AB - OBJECTIVE: The objective of this study was to evaluate the pattern of cognitive disturbances in Nigerian Africans with newly diagnosed epilepsy, prior to onset of drug therapy. METHODS: A total of 60 consecutive patients (mean age 31.6 +/- 17.4, range 14-55 years) presenting with a clinical diagnosis of epilepsy were recruited for the study. Sixty healthy volunteers without a history of epilepsy and who were age-, sex- and level of education matched with the epileptic patients, were recruited as controls. The administration of cognitive tests was done with the FePsy computerized neuropsychological test battery. The tests administered were the visual and auditory reaction times, the continuous performance test and the recognition memory tests to assess mental speed, attention and memory respectively. The means of the cognitive performances of the epileptic patients and controls were statistically compared. RESULTS: Epileptic patients performed worse than the controls across the spectrum of cognitive tasks assessed (P = 0.00001; P < 0.025), with the exception of the beta parameter (response bias) of the vigilance test (P = 0.488; P > 0.025). CONCLUSIONS: The cognitive impairments of short-term memory, psychomotor speed and sustained attention observed in this study are similar to those reported in the literature for patients with epilepsy. The results of this study will be useful in the counseling of patients on their educational, social and vocational needs. PMID- 10830351 TI - Amyotrophic lateral sclerosis. PMID- 10830350 TI - Hydroxyl radical production in the cortex and striatum in a rat model of focal cerebral ischemia. AB - BACKGROUND: Increases in hydroxyl radical production have been used as evidence of oxidative stress in cerebral ischemia/ reperfusion. Ischemia can also induce increased dopamine release from the striatum that may contribute to hydroxyl radical formation. We have compared hydroxyl radical production in the cortex and striatum as an index of oxidative stress in a rat model of focal cerebral ischemia with cortical infarction. METHODS: Using a three vessel occlusion model of focal cerebral ischemia combined with bilateral microdialysis, hydroxylation of 4-hydroxybenzoate (4HB) was continuously monitored in both hemispheres in either the lateral striatum or frontoparietal cortex. The ischemia protocol consisted of one hour equilibration, 30 min of three vessel occlusion, then release of the contralateral common carotid artery (CCA) for 2.5 h. RESULTS: Induction of ischemia resulted in a 30-fold increase in dopamine release in the lateral striatum. Compared to the nonischemic striatum, the ratio of the hydroxylation product 3,4-dihydroxybenzoate (34DHB) to 4HB (trapping agent) in the ipsilateral striatum increased significantly 30 min after ischemia induction. In contrast, during the 30 min of three vessel occlusion there was no increase in the ratio in the cortex. Following the release of the contralateral CCA, the ratio from the ischemic cortex increased significantly compared to sham-operated animals. However, under all circumstances, the 34DHB/4HB ratio was greater in the striatum than in the cortex. CONCLUSION: The increase in the 34DHB/4HB ratio in the lateral striatum coincides with the increased dopamine release suggesting a role for dopamine oxidation in the increased production of hydroxyl radicals. The significant increase in the ratio from the ischemic cortex compared to that from the sham-operated animals is consistent with increased oxidative stress induced by ischemia. However, the lower 34DHB/4HB ratio in the cortex which does not receive dopaminergic innervation compared to the striatum suggests a different mechanism for hydroxyl radical production. Such an alternate mechanism may represent a more toxic oxidative insult that contributes to infarction. PMID- 10830352 TI - Endovascular therapy of a large vertebral artery aneurysm using stent and coils. AB - BACKGROUND: Endovascular therapy is becoming an increasingly popular treatment for cerebral aneurysms. Total angiographic occlusion of small-necked aneurysms (<4 mm) can be obtained in a high percentage of cases. The endovascular treatment of wide-necked or fusiform aneurysms remains a challenge with complete angiographic occlusion reported in <15% of cases. CASE REPORT: We describe the combined use of a flexible coronary stent and platinum coils to treat a wide necked aneurysm of the distal left vertebral artery, in a patient with Grade IV subarachnoid hemorrhage. RESULTS: The procedure was technically successful as the parent artery was protected by the stent while coils were deposited in the aneurysm lumen. Although angiographic aneurysm occlusion was incomplete, the dome was packed with coils. No further hemorrhage has occurred. CONCLUSION: Combined endovascular stent and coil therapy is a promising technique for the treatment of wide-necked cerebral aneurysms. PMID- 10830353 TI - Epilepsy in contemporary fiction: fates of patients. AB - Fictional accounts of epilepsy are of interest because they may convey information on images and public views of epilepsy which are not contained in medical texts. Thus, medical and nonmedical traditions together form the cultural history of epilepsy. Of the numerous possible aspects of epilepsy in fiction, this paper looks especially at the writers' background of knowledge about epilepsy: epilepsy as a handicap and a reason for social rejection, with special reference to epilepsy under the Nazi rule; threats to patients' lives; the motive of the child with epilepsy as a divine child; and epilepsy as a fate, and a reason for distinction. Literary writers may help their readers understand that a person's suffering and fighting a condition like epilepsy very much deserves our attention and sympathy. Without being exclusive, the paper pays special attention to epilepsy in the writings of Canadian authors. PMID- 10830354 TI - Synthetic array measurements of acoustical waves propagating into a water saturated sandy bottom for a smoothed and a roughened interface AB - Measurements of acoustical waves propagating in an unconsolidated water-saturated porous medium that involved the use of a two-dimensional synthetic array technique are presented. From these measurements, wave-front propagation is evaluated using time domain, wave-number frequency and two-dimensional spatial analysis techniques. First, a synthetic array measurement is employed to study sound penetration for a "smoothed" interface condition. This smoothed interface measurement is then compared to an identical synthetic array measurement with a "roughened" interface. A comparison of the smoothed and roughened interface measurements shows enhanced bottom penetration at shallow grazing angles as a result of the roughened interface. At shallow grazing angles, this scattered pressure from the roughened interface has the appearance of a wave front in the water-saturated porous medium that has a virtual wave speed of approximately 1200 m/s. Slower compressional waves in the sandy bottom are not observed. PMID- 10830355 TI - Acoustic scattering from two circular apertures in a thick hard plane AB - A problem of acoustic wave scattering from two circular apertures in a thick hard plane is solved. The Hankel transform and mode matching is used to represent the scattered field in rapidly convergent series. The reflection coefficient, transmission coefficient, and far-zone field are rigorously derived and presented in numerically efficient forms. Numerical computations are performed to illustrate scattering behaviors in terms of aperture geometry. PMID- 10830356 TI - Scale model experiments on the insertion loss of wide and double barriers AB - The insertion loss of wide and double barriers is investigated through scale model experiments. Such configurations appear in outdoor sound propagation problems such as highway noise reduction and community noise control. The Biot Tolstoy-Medwin (BTM) time domain wedge formulation for multiple diffraction [J. Acoust. Soc. Am. 72, 1005-1013 (1982)] is used to predict the acoustic response of an impulsive source. Evaluation of the insertion loss at discrete frequencies is accomplished via the fast Fourier transform (FFT). Good agreement has been found between the BTM model and experimental data for all configurations tested. PMID- 10830357 TI - "Fast" quasilongitudinal sagittally polarized surface waves in layer-substrate structures AB - The propagation of surface sagittally polarized waves on the plated surface of a semi-infinite medium with the phase velocity exceeding that of bulk quasitransverse sagittally polarized waves in the substrate is studied analytically and numerically. Such a surface wave is shown to exist in nonpiezoelectric and piezoelectric composites provided both the sagittal plane and the interface are planes of elastic symmetry of the layer and substrate. In the case of a nonpiezoelectric structure, the "fast" surface wave is one component, involving only the quasi-longitudinal nonuniform mode in the substrate. In piezoelectrics this wave is two-component and incorporates the quasilongitudinal mode and the mode of electrical potential in the substrate. The "fast" wave exists at definite values of the velocity nuF and the layer thickness/wavelength ratio (h/lambda)F. It has been found that one or two surface waves can arise for the same direction of propagation at different nuF and (h/lambda)F, in the case of two surface solutions, both the values of (h/lambda)F being small. The approximate expressions are derived which allow one to elucidate whether the structure under consideration can support the "fast" surface wave and to estimate the corresponding nuF and (h/lambda)F. A comparison is performed of numerical and analytical results. PMID- 10830358 TI - Circular asymmetric Helmholtz resonators AB - A three-dimensional (3D) analytical approach is developed to account for the nonplanar wave propagation in the cavity and neck of "piston-driven" circular asymmetric Helmholtz resonators. The present 3D analytical results are compared with (1) the numerical predictions from the boundary element method (BEM) to evaluate the analytical approach; and (2) the one-dimensional (1D) solution to examine the effect of nonplanar waves at area discontinuity between the neck and the cavity. In order to improve the 1D solution, the end correction is also determined by using the 3D analytical approach. The effect of neck offset on the resonance frequency of circular asymmetric Helmholtz resonators is investigated. Predictions of resonance frequency and transmission loss from the present 3D and corrected 1D analytical approaches are, respectively, identical and close to the BEM results, while the corrected 1D approach provides a better accuracy compared to the 1D solutions with Ingard's correction. Finally, the boundary element method is employed to determine the wave attenuation performance of the "pipe mounted" Helmholtz resonators to examine the effect of multidimensional waves in the vicinity of the main duct and neck junction. PMID- 10830359 TI - Transfer and Green functions based on modal analysis for Lamb waves generation AB - This work presents an easy way to deduce the tensorial transfer and Green functions for Lamb waves generated in isotropic elastic plates. These functions could be applied to obtain the response of each propagating mode in the ensemble of excited modes arising from any sort of pulsed excitation (wedge transducers, lasers, etc.). The transfer function is based on modal analysis development. Not only is it easy to manipulate but also allows the avoidance of laborious calculations for each kind of Lamb waves source. Theoretical predictions are compared with those of Viktorov [I. A. Viktorov, Rayleigh and Lamb Waves (Plenum, New York, 1967)] and with experimental measurements of Lamb waves generated by the wedge-transducer method. PMID- 10830360 TI - Pseudostatic corrections for the forced vibroacoustic response of a structure cavity system AB - The analysis of the coupled vibroacoustic behavior of a structure-cavity system is often performed using the in vacuo structure modes and the rigid cavity modes. Unfortunately, the use of such a modal basis can result in a poor convergence when the high-frequency modes of one of the two subsystems are coupled to the low frequency modes of the other subsystem. This problem is made critical by the lack of a reliable criterion for selecting the number of kept modes for each subsystem. This paper shows that the effect of modal truncation is critical for the convergence of the method and that the convergence can be greatly improved using pseudostatic corrections for both the structure and the cavity. The theory behind the proposed technique is presented together with two generic problems exhibiting strong coupling: (i) an elastic cylindrical cavity filled with air; and (ii) an elastic plate coupled to a rectangular cavity filled with water. PMID- 10830361 TI - Calculation of acoustic radiation using equivalent-sphere methods AB - Among the methods generally used to solve a problem in the domain of acoustic radiation, the equivalent sources method offers an interesting alternative. It consists in replacing the vibrating surface with a distribution of acoustic sources placed inside the structure. The contribution of each source is determined in such a way that the acoustic field radiated by these sources verifies the same boundary conditions on the structure. The number of unknowns in the problem is no longer directly linked to the number of mesh points on the structure, as with boundary elements methods, but to the number of equivalent sources employed in the model. The equivalent source method is therefore of major interest if the acoustic radiation of the structure can be approximated with a sufficiently low number of sources. This paper proposes its application when the equivalent source is a sphere. In this case, the number of unknowns is equal to the number of modes. In contrast to the one-point multipole, the sphere has a surface surrounding a closed volume to express the boundary conditions. Although sphere/multipole equivalence has been demonstrated, the surface of the sphere allows normalization of the functions used, leading to stabilization of the system to be resolved. First, the main acoustic radiation characteristics of a sphere and of the linear system verified by the modal coefficients of an equivalent sphere are presented. The different parameters of the model are then studied: position and radius of the equivalent sphere, truncation of the series, and influence of the spatial sampling (mesh). In the same vein, a second approach is presented. It consists of making each point of the structure correspond to a point of the sphere, and the vibrating field at the surface of the sphere is deduced from that of the structure by simple geometric projection. Results can be obtained very quickly as no matrix inversion is required. The accuracy of the results depends on the distance between the sphere and the structure. Finally, an experimental validation that uses both methods is presented and shows interesting results when the structure is closed, and when its shape is not too far removed from a sphere. PMID- 10830362 TI - Mass, momentum, and energy transfer by the propagation of acoustic solitary waves AB - This paper considers the mass, momentum, and energy transfer accompanied by the propagation of the acoustic solitary wave in a gas-filled tube. As was demonstrated previously [J. Acoust. Soc. Am. 99, 1971-1976 (1996); Phys. Rev. Lett. 83, 4053-4056 (1999)], the propagation of the solitary waves is made possible by connecting a periodic array of Helmholtz resonators axially with the tube. The solitary wave can convey the mass, momentum, and energy steadily with a constant speed that is subsonic but nearly equal to the linear sound speed. It is emphasized that the quantities transferred are of first order in magnitude. Formulating the basic equations in the conservation form, the total amount of the mass, momentum, and energy transferred is obtained by using the solitary-wave solutions. It has the upper bounds determined by the limiting solitary wave, which are proportional to the size of the resonator and the inverse of its natural frequency. In evaluating the energy transfer, a clear distinction should be made between the gas-dynamic energy and the acoustic energy. The former, which contains the first-order quantity whereas the latter begins with the quadratic ones, is to be used to determine the energy transfer in the form of heat (internal energy) associated with the mass transfer. PMID- 10830363 TI - Three-dimensional acoustic scattering from a penetrable layered cylindrical obstacle in a horizontally stratified ocean waveguide AB - In this work, a normal-mode solution is presented for the three-dimensional problem of acoustic scattering from a penetrable horizontally layered cylindrical obstacle in a shallow-water waveguide. The ocean environment around the obstacle is considered horizontally stratified and the bottom is assumed to be rigid. In the general case of depth-dependent sound-speed and density profiles (ssdp) the total acoustic field is calculated numerically, while an analytic solution is obtained in the special case of depth-independent ssdp. Numerical results concerning the transmission loss outside and inside single or double-layered cylindrical structures made of acoustic materials are given for a typical depth dependent ocean environment. Comparisons with the cases of ideally soft and ideally hard cylindrical obstacles [J. Acoust. Soc. Am. 100, 206-218 (1996)] are also made, illustrating the effect of acoustic properties of the obstacle. An important feature, which clearly emerges from the theoretical analysis and the numerical results, is the necessity of including evanescent modes in the calculations, in order to obtain physically meaningful and numerically accurate results. Furthermore, analytical expressions for the scattering cross section of a (penetrable or impenetrable) cylindrical obstacle are derived in terms of the expansion coefficients of the pressure field, and their behavior as frequency increases is numerically investigated. The solution presented in this paper, although addressing a special geometry, provides a means for handling strong discontinuities in both vertical and horizontal directions, and can serve as a benchmark solution to a problem for which no general numerical model exists, i.e., modeling acoustic scattering from a 3D obstacle in a 3D shallow-water waveguide. PMID- 10830364 TI - Time reversal in a waveguide: study of the temporal and spatial focusing AB - Temporal and spatial focusing properties of time-reversal mirrors (TRMs) are studied in a waveguide. The experiments are done using an ultrasonic TRM in an idealized waveguide. The width of the focal spot, and the spatial and temporal sidelobe levels are experimentally and numerically analyzed with respect to the characteristics of the waveguide-TRM system. An algorithm is developed to compute directly in the time domain the time-reversed field. This algorithm is based on the application of the mirror theorem to both the source and the TRM placed in the waveguide. Because time reversal is a stable and robust process, some of the ultrasonic results can be extended to ocean acoustics. Applications to underwater acoustic transmissions as well as ultrasonic medical imaging are discussed. PMID- 10830365 TI - Analysis of resolution for an amplitude-steered array AB - In 1976, Hughes and Thompson introduced the idea of steering the maximum response of a linear array by amplitude weighting the output signals of the elements, thus eliminating the need for time delays or phase-shift networks. Currently that amplitude-steered array concept is being extended to a broadband two-dimensional array that can be used for real-time three-dimensional imaging. In shifting the use of the amplitude-steered array from underwater acoustic communications to imaging, we must consider different issues of the array's performance such as lateral and axial resolution. For the linear amplitude-steered array, we show that both lateral and axial resolution are limited by the length of the array. The dependence of axial resolution on the length of the array is a unique feature of the amplitude-steered array, leading to an interesting tradeoff between lateral and axial resolution. A theoretical basis for the dependence is developed and simulation results are given. PMID- 10830366 TI - A model for longitudinal and shear wave propagation in viscoelastic media AB - Relaxation models fail to predict and explain loss characteristics of many viscoelastic materials which follow a frequency power law. A model based on a time-domain statement of causality is presented that describes observed power-law behavior of many viscoelastic materials. A Hooke's law is derived from power-law loss characteristics; it reduces to the Hooke's law for the Voigt model for the specific case of quadratic frequency loss. Broadband loss and velocity data for both longitudinal and shear elastic types of waves agree well with predictions. These acoustic loss models are compared to theories for loss mechanisms in dielectrics based on isolated polar molecules and cooperative interactions. PMID- 10830367 TI - Weakly localized waves on the corrugated surface of a medium of arbitrary anisotropy AB - The existence of weakly localized surface and pseudosurface waves on the corrugated surface of a medium of arbitrary symmetry is studied. The localized solution originates from an exceptional bulk wave associated with a transonic state of positive or negative curvature. It is shown that the period of corrugation or the frequency of the wave can universally be chosen such that the solution will exist. The range of permissible periods (frequencies) is different for positive and negative curvature transonic states, it being wider near transonic states of positive curvature. The imaginary component of the pseudosurface wave velocity is found to vary as the height of grooves to period of corrugation ratio raised in fourth or sixth power, depending on the transonic state with which the exceptional wave is associated. A simple analytical expression is derived allowing estimations to be made of the penetration depth and the velocity of the localized wave. PMID- 10830368 TI - The quasistatic approximation for a cracked interface between a layer and a substrate AB - Heuristic in nature, the quasistatic approximation (QSA) describes the interaction of ultrasonic waves with imperfect interfaces by modeling the interfacial imperfection as distributions of springs and masses. The QSA does not provide any relationship between the interfacial stiffness constants and the micromechanics of the defects. The aims of this paper are threefold. First, a derivation from first principles of the QSA boundary conditions on a cracked interface is presented. Relationships linking the interfacial constants to the mechanical and geometrical properties of the distributed cracks are also obtained. Second, the stiffness dependence of a cracked interface between a layer and a substrate on the layer thickness is investigated. It is shown that the interfacial stiffnesses cannot be regarded as intrinsic properties of the interface, but they may also depend on the structural properties of the hosting system. Finally, the effect of the thickness dependence of the interfacial stiffnesses on the phase velocity of the lowest mode supported by the layered structure is investigated. PMID- 10830369 TI - Integration of modeling and acoustic microscopy measurements for thin films AB - A model for measuring the V(z) curve by line-focus acoustic microscopy contains the reflectance function of the specimen as a principal component. In this paper the reflectance function has been analyzed for multilayered thin films on a substrate for both fast-on-slow and slow-on-fast systems. The phase velocities of modes of surface acoustic wave propagation and their associated mode reflection coefficients can be obtained from the reflectance function. This information can be used together with estimates of the elastic constants to determine suitable frequency ranges for measuring the V(z) curve. Minimization of the difference between phase velocities obtained from measured and calculated V(z) curves is used to determine the elastic constants. Results are presented for TiN films on M2 high-speed steel substrates. PMID- 10830370 TI - The use of partially measured source data in near-field acoustical holography based on the BEM AB - In applying the conformal near-field acoustical holography (NAH) to actual source identification problems, it is often possible to determine the velocity at certain points of the source surface in advance. This partially known velocity data would reduce the problem size and permit better reconstruction accuracy. In this paper, the effectiveness of using partially measured source data in the conformal NAH is investigated, which uses the boundary element method. A vibro acoustic transfer matrix and measured field pressure data, which is involved with the boundary integral equation, are reorganized in order to deal with the partially measured surface velocities. For a baffled vibrating panel, simulations were performed by varying the number of velocity-known nodes. In addition, the effect of measurement error is investigated for two extreme positioning methods of velocity-known nodes. Without regularization, the reconstructed error can be reduced considerably by employing some of the source data and this error can be further reduced by increasing those surface points. However, the velocity reconstruction error is not reduced substantially when the number of velocity known nodes is less than 30%-40% of the total nodes. The reduction in the reconstruction error is not large if the regularization technique is applied to the restored field. PMID- 10830371 TI - Performance measurements on a thermoacoustic refrigerator driven at high amplitudes AB - Since the power density in a thermoacoustic device is proportional to the square of the acoustic Mach number, there is strong motivation to design thermoacoustic refrigerators to operate at larger pressure amplitudes. Measurements are reported of a modified version of the Space Thermo-Acoustic Refrigerator (STAR), driven at peak-to-mean pressure ratios up to 6%. This pressure ratio corresponds to 30 W of cooling power-five times as large as reported for STAR in 1993. The results of these measurements are compared to a DELTAE computer model of the low-amplitude (linear) performance that matches experimental conditions on a point-by-point basis. It is found that there is a small but measurable deviation in heat pumping power from the power predicted with a linear acoustic computer model at moderate amplitudes. This deviation in heat pumping power at 6% pressure ratio is about 23%. A large disagreement in the acoustic power needed to attain a specific pressure ratio is found between measured data and DELTAE results. An overview of the instrumentation, including a measurement of exhaust heat with an absolute accuracy of 65 mW, is also presented. PMID- 10830372 TI - Thickness shearing vibration of the tangentially polarized piezoelectric ceramic thin circular ring AB - The thickness shearing vibration of the tangentially polarized piezoelectric ceramic thin circular ring is studied. The electrical and mechanical characteristics of the ring are analyzed in detail. The electro-mechanical equivalent circuit, the input electrical impedance, and the frequency equations for the resonance and anti-resonance frequencies are derived. A new concept that is characterized by the cross sectional shape and dimension is presented. An important conclusion is obtained that the thickness shearing vibration of the tangentially polarized piezoelectric ceramic ring is different from the traditional thickness extensional and radial extensional vibrations in that the electro-mechanical conversion coefficient, the electro-mechanical coupling coefficient, and the resonance frequency equation depend not only on the material parameters and the longitudinal dimension, but also on the lateral dimension, such as the cross sectional radius. It is shown that the measured resonance frequencies are in agreement with the theoretically calculated results, and the theoretical relationship between the resonance frequency and the cross sectional radius is also verified. PMID- 10830373 TI - Interaction between wave number pairs for a capped cylinder AB - When acoustic-structure interaction is analyzed according to the wave number based version of the surface variational principle (SVP), considerable computational effort is required to generate each coefficient in the quadratic sum forming the variational quantity. Examination of the wet-surface impedance, which represents the spectrum of pressure amplitudes generated by a specified spectrum of surface velocity amplitudes, reveals that many cross-impedance terms are very small. This suggests that some coefficients need not be computed. The article introduces a priori criteria for selecting the impedance terms to be omitted, based on the supersonic cutoff wave number. Each truncation scheme is assessed by comparing its predictions to the convergent SVP solution. It is shown that, for the nonsymmetric azimuthal harmonics, the field quantities and the radiated power are well predicted if subsonic waves are ignored. In contrast, for the axisymmetric component, substantial errors (6 dB or more) for radiated power arise, unless a broad spectrum of subsonic waves are included in the formulation. The complex power is mostly reactive, being associated with an evanescent field, even for the supersonic spectrum. The study shows that small interactions between subsonic waves in the axisymmetric case can result in the high wave number spectrum being associated with substantial radiation contributions. PMID- 10830374 TI - Acoustic radiation from an elastic baffled rectangular plate covered by a decoupling coating and immersed in a heavy acoustic fluid AB - The vibroacoustic behavior of an elastic, simply supported rectangular plate covered by a locally reacting decoupling layer supporting thickness deformation is presented. The model simulates the vibration and acoustic response of the system immersed in water and subjected to a point force disturbance. A simplified version of the theory is derived in the limiting case of a large decoupling (low mechanical impedance of the layer/high frequency). An appropriate vibratory indicator, representative of the acoustic attenuation provided by the decoupling treatment, and independent of the structure dimensions, is also investigated from the perspective of small-scale laboratory characterization. PMID- 10830375 TI - On reconstruction of acoustic pressure fields using the Helmholtz equation least squares method AB - This paper presents analyses and implementation of the reconstruction of acoustic pressure fields radiated from a general, three-dimensional complex vibrating structure using the Helmholtz equation least-squares (HELS) method. The structure under consideration emulates a full-size four-cylinder engine. To simulate sound radiation from a vibrating structure, harmonic excitations are assumed to act on arbitrarily selected surfaces. The resulting vibration responses are solved by the commercial FEM (finite element method) software I-DEAS. Once the normal component of the surface velocity distribution is determined, the surface acoustic pressures are calculated using standard boundary element method (BEM) codes. The radiated acoustic pressures over several planar surfaces at certain distances from the source are calculated by the Helmholtz integral formulation. These field pressures are taken as the input to the HELS formulation to reconstruct acoustic pressures on the entire source surface, as well as in the field. The reconstructed acoustic pressures thus obtained are then compared with benchmark values. Numerical results demonstrate that good agreements can be obtained with relatively few expansion functions. The HELS method is shown to be very effective in the low-to-mid frequency regime, and can potentially become a powerful noise diagnostic tool. PMID- 10830376 TI - Active control of harmonic sound transmission into an acoustic enclosure using both structural and acoustic actuators AB - This paper describes an analytical and experimental investigation into the active control of harmonic sound transmission in a structural-acoustic coupled system. A rectangular enclosure is considered that has five acoustically rigid walls and a flexible plate on the remaining side through which a harmonic sound wave is transmitted into the enclosure. The control system is designed to globally reduce the sound field inside the enclosure, and the roles of structural and acoustic actuators are of particular interest. Three control configurations, classified by the type of actuators, are compared and discussed. They are: (i) use of a single point-force actuator, (ii) use of a single acoustic piston source, and (iii) simultaneous use of both a point-force actuator and an acoustic piston source. It is shown both analytically and experimentally that the point-force actuator is effective in controlling plate-dominated modes while the acoustic source is effective in controlling cavity-dominated modes. Since the transmitted sound field is governed by both plate- and cavity-dominated modes, the hybrid use of both types of actuators is shown to be a desirable configuration for the active control of sound transmission into a structural-acoustic coupled system. PMID- 10830377 TI - Effects on sleep disturbance of changes in aircraft noise near three airports. AB - Field measurements were conducted of potential sleep disturbance associated with changes in nighttime aircraft noise exposure near three airports. One study was conducted near Stapleton International Airport (DEN) and Denver International Airport (DIA) in anticipation of the closure of the former and opening of the latter. Sleep behavior was monitored in 57 homes located near runway ends at the two airports. A second study was conducted in the vicinity of DeKalb-Peachtree Airport (PDK), a large general aviation airport that expected increased nighttime flight operations due to the Olympic Games in July and August of 1996. Similar methods of measuring nighttime noise levels and sleep disturbance in the two studies were maintained over the course of 2717 and 686 subject-nights of observations, respectively. No major differences in noise-induced sleep disturbance were observed as a function of changes in nighttime aircraft noise exposure. PMID- 10830378 TI - Surface area measurement utilizing an acoustic bridge AB - A new method is proposed for measuring the surface area of an object. The acoustic conductance of a cavity is proportional to the surface area of the cavity inner wall. The surface area of an object thus can be known from the measurement of the acoustic impedance of a chamber in which the object is placed. In order to measure the acoustic impedance accurately; the proposed method employs the acoustic bridge technique. The experimental device is composed of the following elements so arranged that their electric equivalents form a bridge circuit: a measuring chamber in which an object under test is placed and whose volume can be adjusted; a reference chamber whose inner surface area can be varied; a loudspeaker (the signal source) mounted between the two chambers; and a bypass channel at the midpoint of which a microphone (the null detector) is installed. This bridge balances when the volume and the inner surface area of each chamber become equal. The surface area of the object can then be known from the inner surface area of the reference chamber. Several experiments were performed with this device and the success of the proposed method was verified. PMID- 10830379 TI - Inverse problem solution techniques as applied to indirect in situ estimation of fish target strength. AB - In situ indirect methods of fish target strength (TS) estimation are analyzed in terms of the inverse techniques recently applied to the problem in question. The solution of this problem requires finding the unknown probability density function (pdf) of fish target strength from acoustic echoes, which can be estimated by solving the integral equation, relating pdf's of echo variable, target strength, and beam pattern of the echosounder transducer. In the first part of the paper the review of existing indirect in situ TS-estimation methods is presented. The second part introduces the novel TS-estimation methods, viz.: Expectation, Maximization, and Smoothing (EMS), Windowed Singular Value Decomposition (WSVD), Regularization and Wavelet Decomposition, which are compared using simulations as well as actual data from acoustic surveys. The survey data, acquired by the dual-beam digital echosounder, were thoroughly analyzed by numerical algorithms and the target strength and acoustical backscattering length pdf's estimates were calculated from fish echoes received in the narrow beam channel of the echosounder. Simultaneously, the estimates obtained directly from the dual-beam system were used as a reference for comparison of the estimates calculated by the newly introduced inverse techniques. The TS estimates analyzed in the paper are superior to those obtained from deconvolution or other conventional techniques, as the newly introduced methods partly avoid the problem of ill-conditioned equations and matrix inversion. PMID- 10830380 TI - The matched-phase coherent multi-frequency matched-field processor AB - Coherent multi-frequency matched-field processing is investigated using a matched phase coherent matched-field processor. Its main difference from previous coherent processors is that the relative phases of the Fourier components contained within the recorded signal are not assumed to be known a priori. Rather they are considered free parameters that can be determined using a global functional minimization algorithm. Additionally, this processor uses only the cross-frequency terms, making it less susceptible to the detrimental effects of ambient noise; in one example, this processor shows a five decibel improvement over a similar coherent processor. Along with its increased sensitivity with respect to the broadcast source levels, this coherent processor exhibits superior range resolution as compared with multi-frequency incoherent processors, due to the cross-frequency interference of the vertical eigenmodes. Within this work we explore the efficacy of the algorithms used to determine the relative phases along with the performance of the matched-phase coherent processor itself, performed within the context of data collected during an event from the SWellEx 96 experiment. Performance comparisons between this processor, an incoherent processor, and another coherent processor are demonstrated using this data set. PMID- 10830381 TI - Use of higher order statistics in source signature estimation AB - Higher order statistical blind deconvolution methods are implemented for use in removing multipath distortion from passively received underwater acoustic transient signals. Using single channel data and simulations, it is demonstrated that a fourth order method based on cumulant maximization can work well if the associated multipath Green's function is sufficiently "sparse." The iterative method is parameterized by filter length, and while there is a range of values at which the best solutions are obtained with conventional convergence criteria, useful solutions exist across a much broader range of filter lengths if the iterations are not always allowed to proceed to convergence. The fourth order objective functional is generalized to arbitrary order, and the method is shown to also produce good results for the third order objective functional. PMID- 10830382 TI - Frequency responses of two- and three-tone distortion product otoacoustic emissions in Mongolian gerbils. AB - The frequency responses of distortion product otoacoustic emission (DPOAEs) were investigated in adult Mongolian gerbils. The main goal was to investigate in this species the extent to which DPOAE measurements might be useful in estimating cochlear frequency-tuning characteristics. Specifically, this study investigated the parameter space for generation of DPOAEs to determine those regions, if any, where the emission responses gave "simple" frequency responses, i.e., responses similar in form to typical neural responses. At the same time, it was desired to determine in this species the existence, extent, and nature of the more complex three-tone emission frequency responses as observed in some other species [e.g., Martin et al., Hearing Res. 136, 105-123 (1999)]. In the present work, two-tone frequency response curves (f2/f1 ratio functions) were obtained by varying the lower frequency, f1, while holding the f2 frequency and both amplitudes (L1, L2) constant. Only for frequencies, f2, near 8 kHz did the response at the emission frequency, 2 f1-f2, form a simple, relatively broad peak. At all lower frequencies, the two-tone frequency response curve was typically complex and composed of multiple peaks. In comparison, three-tone frequency responses were constructed by fixing the primary stimulus pair (f1, f2) and varying a third tone widely in frequency (f3) and intensity (L3). Points in f3 and L3 which caused a criterion reduction in primary emission amplitude (at 2 f1-f2) were used to construct emission suppression tuning curves (STCs). Only for primary frequencies, f2, at 8 kHz and above were the emission STCs found to be simple, with shapes similar to neural frequency-tuning curves. At lower primary frequencies, particularly for relatively low primary frequency ratios (low f2/f1), three-tone responses were very complex. This complex response usually included a region of anomalous suppression in which very low suppression levels (L3) could result in significant decreases in the primary emission amplitude, often exceeding 12 dB. Regions of such anomalous suppression were typically observed under the following conditions: (1) for all f2 frequencies from 0.5 to 4 kHz; (2) for f3 frequencies between 1.4 and 8 kHz; (3) i.e., for f3 frequencies 1 3 octaves above the primary frequency, f2; (4) at L3 levels often 10 dB lower or more than the usual "best frequency" threshold, i.e., even lower than the relative minimum threshold found near the primary stimulus frequencies; (5) exhibiting sharp amplitude decreases often accompanied by emission phase shifts of about 180 deg; (6) present in both cubic emissions (2 f1-f2 and 2 f2-f1); (7) to be less extreme at larger primary stimulus frequency ratios (larger f2/f1); and (8) less extreme at larger intensity ratios (larger L1/L2). Because of the anomalous behavior at f2 frequencies below 8 kHz, "simple" emission STCs were typically only obtainable, if at all, near the extreme boundaries of the parameter space giving measurable emission amplitudes. PMID- 10830383 TI - Effects of acoustic trauma on acoustic enhancement of electrically evoked otoacoustic emissions. AB - Moderate acoustic trauma results in decreased cochlear sensitivity and frequency selectivity. This decrease is believed to be caused by damage to the cochlear amplifier that is associated with outer hair cells (OHCs) and their nonlinear electromechanical characteristics. A consequence of OHC nonlinearity is the acoustic enhancement effect, in which low-frequency electrically evoked otoacoustic emissions are enhanced by a simultaneous tone. The present study found that acoustic trauma reduced the acoustic enhancement effect and this reduction is correlated with the N1 threshold at the electrode site. This result is consistent with the theory that trauma affects the mechanoelectric transduction process, thus affecting cochlear mechanical nonlinearity. Acoustic trauma also reduced the cochlear microphonic in a way that suggests that the number of functioning tension-gated channels and the stiffness of the gating springs were decreased. In some cases, the electromechanical transduction process was also found to be affected by acoustic trauma. PMID- 10830384 TI - Auditory-nerve-fiber responses to high-level clicks: interference patterns indicate that excitation is due to the combination of multiple drives. AB - There has been no systematic study of auditory-nerve-fiber (ANF) responses to high-level clicks despite the advantages of clicks in revealing the natural resonances of a system. Cat single ANFs were studied using clicks up to 120 dB pSPL. Peri-stimulus-time (PST) histograms of responses were corrected for refractory effects, and compound PST (cPST) histograms were formed from rarefaction- and condensation-click PSTs. At low levels the responses followed the classic picture with each cPST appearing to be from a single resonant system followed by low-pass filtering that reduces high-frequency synchrony. In fibers across all characteristic frequencies, there were significantly different patterns at high click levels including several nonclassic features and "phase reversals," i.e., a peak in the rarefaction-click PST at low levels was replaced at high levels by a peak at the same latency in the condensation-click PST. There were two separate regions of nonclassic features and phase reversals, which indicates that auditory-nerve fibers are excited by the combination at some stage in the cochlea of at least three excitation drives derived from the acoustic stimulus. These data support the interpretation that the cochlear partition vibrates in multiple resonant modes with each mode producing one excitation drive and that the mix of modes varies with sound level. PMID- 10830385 TI - The case of the missing pitch templates: how harmonic templates emerge in the early auditory system. AB - Periodicity pitch is the most salient and important of all pitch percepts. Psychoacoustical models of this percept have long postulated the existence of internalized harmonic templates against which incoming resolved spectra can be compared, and pitch determined according to the best matching templates [J. Goldstein, J. Acoust. Soc. Am. 54, 1496-1516 (1973)]. However, it has been a mystery where and how such harmonic templates can come about. We present here a biologically plausible model for how such templates can form in the early stages of the auditory system. The model demonstrates that any broadband stimulus, including noise and random click trains, suffices for generating the templates, and that there is no need for any delay lines, oscillators, or other neural temporal structures. The model consists of two key stages: cochlear filtering followed by coincidence detection. The cochlear stage provides responses analogous to those recorded in the auditory nerve and cochlear nucleus. Specifically, it performs moderately sharp frequency analysis via a filterbank with tonotopically ordered center frequencies (CFs); the rectified and phase locked filter responses are further enhanced temporally to resemble the synchronized responses of cells in the cochlear nucleus. The second stage is a matrix of coincidence detectors that compute the average pairwise instantaneous correlation (or product) between responses from all CFs across the channels. Model simulations show that for any broadband stimulus, a degree of high coincidence occurs among cochlear channels that are spaced precisely at harmonic intervals. Accumulating coincidences over time results in the formation of harmonic templates for all fundamental frequencies in the phase-locking frequency range. The model accounts for the critical role played by three subtle but important factors in cochlear function: the nonlinear transformations following the filtering stage, the rapid phase shifts of the traveling wave near its resonance, and the spectral resolution of the cochlear filters. Finally, we discuss the physiological correlates and location of such a process and its resulting templates. PMID- 10830386 TI - A model of the perceptual asymmetry between peaks and troughs of frequency modulation. AB - Pitch discrimination at peaks of frequency modulation is better than at troughs [L. Demany and K. I. McAnally, J. Acoust. Soc. Am. 96, 706-715 (1989)]. A similar asymmetry emerges within a time-domain pitch perception model based on autocorrelation. The model requires the following assumptions: (a) The neural discharge patterns must be temporally sharpened to a single narrow pulse per period (possibly by neural convergence within the cochlear nucleus). (b) Autocorrelation must be implemented as a cross correlation between the neural pulse train and a delayed pulse train convolved with a short kernel function. This kernel function must be asymmetric in time. (c) Pitch discrimination must rely on higher-order modes of the autocorrelation function. This particular implementation of the autocorrelation model produces modes that are sharper for peaks than for troughs, and thus accounts for the pitch discrimination asymmetry observed experimentally. As a by-product it can account for "hyperacute" discrimination observed at peaks of triangular modulation. PMID- 10830387 TI - Frequency discrimination in budgerigars (Melopsittacus undulatus): effects of tone duration and tonal context. AB - Studies of frequency resolving power in budgerigars (Melopsittacus undulatus) have shown that this species has excellent discrimination abilities for both simple and complex sounds falling in the region of 2 to 4 kHz--the frequency range of their contact call. In four experiments, frequency discrimination by budgerigars of short tones similar to elements found in the contact call was examined. Frequency difference limens (FDLs) for simple pure tones at 2.86 kHz were constant for tone durations above 20 ms but higher for shorter tones. Budgerigars generally showed larger FDLs for shorter duration 1-, 2-, and 4-kHz pure tones. FDLs in budgerigars for 20-ms tones embedded in a sequence of six other tones were similar to FDLs measured for tones of the same frequency presented in isolation. Moreover, there was no effect of introducing trial-by trial variation in the location of the frequency change in the seven-tone complexes for budgerigars, a condition for which humans showed a large decrement in performance. Taken together, these results suggest budgerigars possess enhanced spectral resolving power for short duration pure tones when they are embedded in contact call-like tonal patterns. PMID- 10830388 TI - Feedback path variability modeling for robust hearing aids. AB - Acoustic feedback is a common problem in hearing aids with vented earmolds. Hearing aids designed to work under normal conditions become unstable when the feedback path varies under changing conditions. A comprehensive study of the variability of the feedback path under various conditions and for different users is presented in this paper. A multiplicative uncertainty bound widely used in H infinity robust control is suggested to model the variations, which is then used to formulate a robust stability condition for the hearing aid. The upper limit of the closed-loop acoustic gain of the hearing aid for maintaining robust stability is also derived. Examples of robust constant amplification hearing aids, which maintain stability in the face of the given variations in the feedback path, are presented. The robust stability condition is also suggested as a tool to design more robust digital, hearing aids. PMID- 10830389 TI - Experience with a yes-no single-interval maximum-likelihood procedure. AB - The report in 1993 by Green [J. Acoust. Soc. Am. 93, 2096-2105 (1993)] describing the application of a new psychophysical method requiring few trials and little time to measure auditory thresholds has generated considerable interest among experimentalists. The procedure uses a single-interval stimulus presentation, requests a yes-no decision by subjects, and implements a maximum-likelihood calculation to determine the next trial stimulus level within an adaptive track, as well as the final threshold estimate. Data are presented here describing separate experiences with this procedure in two laboratories in both detection and discrimination tasks. Issues addressed include comparisons with more traditional psychophysical methods, variability in threshold estimates, experimental time required, and possible minor modifications to improve the basic procedure. Results using this procedure are comparable in terms of variability of estimates to those emerging from more lengthy procedures. However, because it may be difficult for some listeners to maintain a consistent criterion and because attentional lapses may be costly, experimenters must be willing to monitor performance closely and repeat some tracks in cases where excessively high variability is noted. Further, this procedure may not be suitable for tasks for which the form of the psychometric function is not well-established. Modifications allowing a variable slope parameter in the maximum-likelihood evaluations of psychometric functions may be of benefit. PMID- 10830390 TI - Phonological conditioning of peak alignment in rising pitch accents in Dutch. AB - This paper deals with the factors that influence the alignment of F0 movements with phonetic segments. It reports two experiments on the alignment of rising prenuclear pitch accents in Dutch. In experiment 1, it is shown that the final peak of the rise is aligned at the end of the vowel if the accented syllable contains a long vowel, but during the following consonant if the accented syllable contains a short vowel. The beginning of the rise is consistently aligned at the beginning of the accented syllable. Experiment 2 attempts to distinguish between two explanations for this finding: (1) a durational account, in which the F0 rise takes a certain amount of time and overruns into the following consonant if the vowel is short; and (2) a structural account, in which the peak of the rise is seen as a tonal target aligned with the end of the syllable (which is structurally earlier for long vowels than for short vowels). The data partially support both accounts. There is an alignment difference despite a lack of durational difference, which supports the structure-based account. However, the effect is reduced compared to experiment 1, showing that time pressure may work against the ideal alignment. PMID- 10830391 TI - Neurophysiologic correlates of cross-language phonetic perception. AB - This study examined neurophysiologic correlates of the perception of native and nonnative phonetic categories. Behavioral and electrophysiologic responses were obtained from Hindi and English listeners in response to a stimulus continuum of naturally produced, bilabial CV stimuli that differed in VOT from -90 to 0 ms. These speech sounds constitute phonemically relevant categories in Hindi but not in English. As expected, the native Hindi listeners identified the stimuli as belonging to two distinct phonetic categories (/ba/ and /pa/) and were easily able to discriminate a stimulus pair across these categories. On the other hand, English listeners discriminated the same stimulus pair at a chance level. In the electrophysiologic experiment N1 and MMN cortical evoked potentials (considered neurophysiologic indices of stimulus processing) were measured. The changes in N1 latency which reflected the duration of pre-voicing across the stimulus continuum were not significantly different for Hindi and English listeners. On the other hand, in response to the /ba/-/pa/ stimulus contrast, a robust MMN was seen only in Hindi listeners and not in English listeners. These results suggest that neurophysiologic levels of stimulus processing reflected by the MMN and N1 are differentially altered by linguistic experience. PMID- 10830392 TI - Speech recognition with reduced spectral cues as a function of age. AB - Adult listeners are able to recognize speech even under conditions of severe spectral degradation. To assess the developmental time course of this robust pattern recognition, speech recognition was measured in two groups of children (5 7 and 10-12 years of age) as a function of the degree of spectral resolution. Results were compared to recognition performance of adults listening to the same materials and conditions. The spectral detail was systematically manipulated using a noise-band vocoder in which filtered noise bands were modulated by the amplitude envelope from the same spectral bands in speech. Performance scores between adults and older children did not differ statistically, whereas scores by younger children were significantly lower; they required more spectral resolution to perform at the same level as adults and older children. Part of the deficit in younger children was due to their inability to utilize fully the sensory information, and part was due to their incomplete linguistic/cognitive development. The fact that young children cannot recognize spectrally degraded speech as well as adults suggests that a long learning period is required for robust acoustic pattern recognition. These findings have implications for the application of auditory sensory devices for young children with early-onset hearing loss. PMID- 10830393 TI - An investigation of current models of second language speech perception: the case of Japanese adults' perception of English consonants. AB - This study reports the results of two experiments with native speakers of Japanese. In experiment 1, near-monolingual Japanese listeners participated in a cross-language mapping experiment in which they identified English and Japanese consonants in terms of a Japanese category, then rated the identifications for goodness-of-fit to that Japanese category. Experiment 2 used the same set of stimuli in a categorial discrimination test. Three groups of Japanese speakers varying in English-language experience, and one group of native English speakers participated. Contrast pairs composed of two English consonants, two Japanese consonants, and one English and one Japanese consonant were tested. The results indicated that the perceived phonetic distance of second language (L2) consonants from the closest first language (L1) consonant predicted the discrimination of L2 sounds. In addition, this study investigated the role of experience in learning sounds in a second language. Some of the consonant contrasts tested showed evidence of learning (i.e., significantly higher scores for the experienced than the relatively inexperienced Japanese groups). The perceived phonetic distance of L1 and L2 sounds was found to predict learning effects in discrimination of L1 and L2 sounds in some cases. The results are discussed in terms of models of cross-language speech perception and L2 phonetic learning. PMID- 10830394 TI - Construction and interpretation of equal-tempered scales using frequency ratios, maximally even sets, and P-cycles. AB - Using recent developments in music theory, which are generalizations of the well known properties of the familiar 12-tone, equal-tempered musical scale, an approach is described for constructing equal-tempered musical scales (with "diatonic" scales and the associated chord structure) based on good-fitting intervals and a generalization of the modulation properties of the circle of fifths. An analysis of the usual 12-tone equal-tempered system is provided as a vehicle to introduce the mathematical details of these recent music-theoretic developments and to articulate the approach for constructing musical scales. The formalism is extended to describe equal-tempered musical scales with nonoctave closure. Application of the formalism to a system with closure at an octave plus a perfect fifth generates the Bohlen-Pierce scale originally developed for harmonic properties similar to traditional chords but without the perceptual biases of these familiar chords. Subsequently, the formalism is applied to the group-theory-based 20-fold microtonal system of Balzano. It is shown that with an appropriate choice of nonoctave closure (6:1 in this case), determined by the formalism combined with continued fraction analysis, that this group-theoretic generated system may be interpreted in terms of the frequency ratios 21:56:88:126. Although contrary to the spirit of the group-theoretic approach to generating scales, this analysis may be applicable for discovering the ratio basis of unusual tunings common in non-Western music. PMID- 10830395 TI - Optimizaition of a low-frequency ultrasonic technique to monitor the change in physical states in viscoelastic media: gelation process AB - A new low-frequency ultrasonic device (50-100 kHz) in highly sharpened end sensors that behave as point sources were examined. The application of this new ultrasonic technique with two sensors coupled in the near field is to explore the relations between the physical properties measured through the evolution of the wave time of flight and structural changes during gel formation which is related to two factors: the ambient temperature and the mechanical resistance of the medium. The network evolution was interpreted by an approach based on the Flory model. The physical significance of this model was shown through a series of experiments using a low-frequency ultrasonic technique. Response curves demonstrate the different stages during gel formation. PMID- 10830396 TI - One-hydrophone method of estimating distance and depth of phonating dolphins in shallow water. AB - Previous attempts at localizing cetaceans have generally used multiple hydrophone arrays and multichannel recording systems. In this paper, a low-budget localization technique using only one hydrophone is described. The time delays of the signals traveling via the surface and bottom reflection paths to the hydrophone, relative to the direct signal, are used to calculate the distance and the depth of a phonating animal. Only two additional measures, the depth of the bottom and hydrophone, have to be taken. The method requires relatively shallow waters and a flat bottom surface. Echolocating and burst pulsing Hawaiian spinner dolphins (Stenella longirostris) at the Waianae coast of Oahu, Hawaii, were localized over different bottom substrates. A tracking range of up to 100 m was achieved. The accuracy of the method is estimated by the total error differential technique. The relative distance estimation error is below 35% and the absolute depth error below 0.7 m, so that the location method is sufficiently precise for examining source levels in our study area. Because of its simplicity, the method ideally complements sound recordings and visual sightings of marine mammals and could lead to a better understanding of the nature and use of click trains by dolphins. PMID- 10830397 TI - Classification of electronically generated phantom targets by an Atlantic bottlenose dolphin (Tursiops truncatus). AB - Animal behavior experiments require not only stimulus control of the animal's behavior, but also precise control of the stimulus itself. In discrimination experiments with real target presentation, the complex interdependence between the physical dimensions and the backscattering process of an object make it difficult to extract and control relevant echo parameters separately. In other phantom-echo experiments, the echoes were relatively simple and could only simulate certain properties of targets. The echo-simulation method utilized in this paper can be used to transform any animal echolocation sound into phantom echoes of high fidelity and complexity. The developed phantom-echo system is implemented on a digital signal-processing board and gives an experimenter fully programmable control over the echo-generating process and the echo structure itself. In this experiment, the capability of a dolphin to discriminate between acoustically simulated phantom replicas of targets and their real equivalents was tested. Phantom replicas were presented in a probe technique during a materials discrimination experiment. The animal accepted the phantom echoes and classified them in the same manner as it classified real targets. PMID- 10830398 TI - Kirchhoff evaluation of scattered elastic wavefields in anisotropic media PMID- 10830399 TI - The noise emitted from vehicles at roundabouts PMID- 10830400 TI - Effect of membrane motor on the axial stiffness of the cochlear outer hair cell. PMID- 10830401 TI - Time-frequency representation of Lamb waves using the reassigned spectrogram PMID- 10830402 TI - An integrated printed circuit board (PCB) microphone PMID- 10830403 TI - Selective attention to the parameters of a physically informed sonic model PMID- 10830404 TI - Episodic movement disorders as channelopathies. PMID- 10830405 TI - Limited-time exposure to mitochondrial toxins may lead to chronic progressive neurodegenerative diseases. PMID- 10830406 TI - Scientific position paper of the Movement Disorder Society evaluation of surgery for Parkinson's disease. Task Force on Surgery for Parkinson's Disease of the American Academy of Neurology Therapeutic and Technology Assessment Committee. PMID- 10830407 TI - Cost-effectiveness analysis in Parkinson's disease: determining the value of interventions. PMID- 10830408 TI - Research goals in progressive supranuclear palsy. First International Brainstorming Conference on PSP. PMID- 10830409 TI - Relationship among nigrostriatal denervation, parkinsonism, and dyskinesias in the MPTP primate model. AB - Presynaptic denervation is likely to play an important role in the pathophysiology of dyskinesias that develop after levodopa administration to patients with Parkinson's disease. In this study, the thresholds of nigrostriatal damage necessary for the occurrence of parkinsonism and levodopa-induced involuntary movements were compared in squirrel monkeys lesioned with 1-methyl-4 phenyl-1,2,3,6-tetrahydropyridine (MPTP). Animals treated with a regimen of MPTP that caused parkinsonism displayed > or =95% striatal dopamine depletion, 90% reduction of striatal dopamine uptake sites, and 70% nigral neuronal loss. Levodopa administration ameliorated the parkinsonian signs of these monkeys but also induced dyskinesias. A separate group of animals was treated with a milder MPTP regimen that caused 60%-70% striatal dopamine depletion, a 50% decrease in dopamine transporter, and 40% loss of dopaminergic nigral neurons. While these monkeys displayed no behavioral signs of parkinsonism, they all became dyskinetic after levodopa administration. The priming effect of levodopa, that is, the recurrence of dyskinesias in animals previously exposed to the drug, was compared in severely versus mildly lesioned monkeys. When severely injured parkinsonian animals underwent a second cycle of levodopa treatment, they immediately and consistently developed involuntary movements. In contrast, the recurrence of dyskinesias in primed monkeys with a partial nigrostriatal lesion required several levodopa administrations and remained relatively sporadic. The data indicate that moderate nigrostriatal damage which does not induce clinical parkinsonism predisposes to levodopa-induced dyskinesias. Once dyskinesias have been induced, the severity of denervation may enhance the sensitivity to subsequent levodopa exposures. PMID- 10830410 TI - Early detection of probable idiopathic Parkinson's disease: I. Development of a diagnostic test battery. AB - We developed a test battery as an inexpensive and objective aid for the early diagnosis of idiopathic Parkinson's disease (iPD) and its differential diagnoses. The test battery incorporates tests of motor function, olfaction, and mood. In the motor task, a wrist flexion-and-extension task to different targets, movement velocities were recorded. Olfaction was tested with the University of Pennsylvania Smell Identification Test. Mood was assessed with the Beck Depression Inventory. An initial regression model was developed from the results of 19 normal control subjects and 18 patients with early, mild, probable iPD. Prospective application to an independent validation set of 122 normal control subjects and 103 patients resulted in an 88% specificity rate and 69% sensitivity rate, with an area under the Receiver Operator Characteristic curve of 0.87. PMID- 10830411 TI - Early detection of probable idiopathic Parkinson's disease: II. A prospective application of a diagnostic test battery. AB - We developed a battery of tests, incorporating motor function, olfaction, and mood, that distinguishes between early mild, probable idiopathic Parkinson's disease (iPD), and normal control subjects. Here, we determined the PD Battery's ability to prospectively identify which patients with symptoms and signs suggestive of, but not yet diagnostic of, iPD who would subsequently be diagnosed as having iPD. Of 212 enrolled patients, 194 have been followed for at least 1 year. Among these 194, 59 were diagnosed with iPD, 40 as neurologically normal, and the remaining 106 with other neurologic conditions such as essential tremor. The test battery was 92% specific and 68% sensitive for iPD. The area under the Receiver Operating Characteristics curve was 0.88 for predicting which participants would subsequently be diagnosed as having iPD or as being neurologically normal. PMID- 10830412 TI - Differential progression of motor impairment in levodopa-treated Parkinson's disease. AB - OBJECTIVE: To monitor comparative progression of clinical impairment over 4 years in patients with Parkinson's disease (PD) who present on levodopa at two different levels of Hoehn and Yahr (HY) stages, II and III. BACKGROUND: The rate of clinical impairment progression in patients with PD being treated with levodopa has not been studied in detail using current, standardized assessment tools. Sample size estimates for all levodopa adjunctive treatment studies and proper definition of study groups require a solid estimate of longitudinal motor impairment progression. DESIGN/METHODS: From our computer database, we identified two groups of patients with PD being treated with levodopa based on their initial HY stage at presentation to our center (II or III). Fifty randomly selected subjects in each stage were monitored in the ON state with annual Unified Parkinson's Disease Rating Scale (UPDRS) motor scores, dyskinesia ratings, and antiparkinsonian medication doses using a repeated measures analysis of variance. RESULTS: The stage II and stage III subjects had similar disease duration. In stage II subjects, parkinsonian impairment was maintained without progression over 4 years, but in association with significantly higher dyskinesia scores and dopaminergic medication doses. In stage III subjects, UPDRS motor scores deteriorated despite more medication and increased dyskinesias. Of the established six factors comprising the UPDRS motor scale, bradykinesia accounted for the increased impairment. Initial UPDRS motor score and disease duration did not influence progression of motor impairment. CONCLUSION: In subjects with similar disease duration, progression of PD motor impairment differs significantly between stage II and stage III subjects over 4 years. Whereas in stage II subjects, parkinsonian impairment can be stabilized at the expense of increased dyskinesia and dopaminergic drugs, once subjects reach stage III, motor impairment progresses. Power estimates and sample size calculations for these groups of patients should be calculated separately. PMID- 10830413 TI - Time course of loss of clinical benefit following withdrawal of levodopa/carbidopa and bromocriptine in early Parkinson' s disease. AB - Putative neuroprotective agents for Parkinson's disease can be assessed in untreated patients using progression of clinical disability as an index of disease progression. To avoid the confound associated with symptomatic therapy, progression of the underlying disease can be assessed by evaluating the progression of clinical disability from an untreated baseline to a final visit following wash-out of symptomatic medication. In this type of analysis it is critical to use a washout of sufficient duration to ensure elimination of symptomatic effects. To assess the time course of resolution of symptomatic effects, we evaluated 31 patients at days 1, 8, and 15 following discontinuation of levodopa/carbidopa and bromocriptine. Mean total Unified Parkinson's Disease Rating Scale scores (+/- standard error) increased (worsened) by 7.4+/-1.5 from day 1 to day 15 (p <0.0001), 4.5+/-1.2 from day 1 to day 8 (p = 0.0009), and 2.9+/-1.0 from day 8 to day 15 (p = 0.01). We conclude that a wash-out of at least 2 weeks is required to eliminate the symptomatic effects of levodopa/carbidopa and bromocriptine in patients with early Parkinson's disease. PMID- 10830414 TI - Gender differences in the frequency and treatment of behavior problems in Parkinson's disease. SAGE Study Group. Systematic Assessment and Geriatric drug use via Epidemiology. AB - OBJECTIVE: To determine gender differences in the prevalence, survival rates, and management of noncognitive behavioral problems of patients with Parkinson's disease (PD) in nursing homes (NH). METHODS: We performed an observational study on 24,402 residents with PD using the Systematic Assessment and Geriatric drug use via Epidemiology (SAGE) database collected from the Minimum Data Set on a cross-section of over 400,000 NH residents in five US states. Gender differences in behavior were used to predict differences in pharmacologic and nonpharmacologic therapies using logistic regression. Similar analyses were done to evaluate gender differences in 1-year survival rates among patients with PD with and without behavioral problems. RESULTS: 36% of men and 33% of women exhibited behavioral problems. Wandering, verbal and physical abusiveness, and inappropriate behavior tended to be more common in men, especially among PD residents with severe cognitive impairment. Hallucinations and delusions were equally prevalent between genders and depressive symptoms were more common in women. Regardless of behavioral manifestation, men were more likely to receive antipsychotic drugs, whereas women were more likely to receive antidepressants. This gender difference in treatment was also widest among the severely demented group. Although women lived longer, no difference in survival curves were noted between PD residents with and without behavioral problems. CONCLUSION: Gender appears to play an important role in determining the frequency and treatment of behavioral problems of NH residents with PD. PMID- 10830415 TI - Illness impact and adjustment to Parkinson's disease: before and after treatment with tolcapone. AB - Research on the impact of disease and treatment on health status and quality of life in patients with movement disorders is limited. We studied quality of life in 46 patients with Parkinson's disease (PD) to determine whether the impact of illness and psychosocial adjustment to illness were improved by 42 days of adjunctive therapy with tolcapone (50 mg, 200 mg, or 400 mg three times a day). This study was conducted in parallel with a double-blind, placebo-controlled, dose-response study of the safety and efficacy of tolcapone in combination with levodopa/carbidopa therapy. Only a subset of individuals from the larger study participated. In the quality of life study, illness impact and adjustment to illness were measured subjectively by the Sickness Impact Profile (SIP) and the Psychosocial Adjustment to Illness Scale-Self-Report (PAIS-SR). Patient ratings of total illness impact (p = 0.003), physical illness impact (p = 0.05), and psychosocial illness impact (p = 0.007) improved significantly in individuals receiving tolcapone compared with those receiving placebo. There was no statistically significant difference in adjustment to illness when the tolcapone and placebo groups were compared; however, 17 of 21 adjustment to illness indicators showed improvement. PMID- 10830416 TI - Accurate differentiation of parkinsonism and essential tremor using visual assessment of [123I]-FP-CIT SPECT imaging: the [123I]-FP-CIT study group. AB - OBJECTIVE: To evaluate whether visual assessment of [123I]-FP-CIT (DaTSCAN, Nycomed Amersham, plc) single photon emission computerized tomography (SPECT) images can differentiate between parkinsonism and essential tremor (ET). METHODS: [123I]-FP-CIT SPECT imaging was conducted in a six-center study of 158 patients with a clinical diagnosis of parkinsonism compared with 27 ET cases and 35 healthy volunteers. Striatal uptake of the radioligand was graded normal or abnormal, and abnormal images were further graded to three levels of severity. An institutional read whereby each center visually assessed the images blinded to the clinical data and a consensus blinded read by a panel of five was undertaken. RESULTS: The institutional reading scored 154 of 158 cases of parkinsonism abnormal, all 27 cases of ET as normal, and 34 of 35 healthy volunteers as normal compared with the consensus blinded read scoring 150 cases of parkinsonism as abnormal, 25 ET cases as normal, and 33 healthy volunteers as normal. Sensitivity for the clinical diagnosis of parkinsonism was 97% and specificity for ET was 100% for the institutional read, whereas sensitivity was 95% and specificity 93% for the consensus blinded read. Semiquantitative analysis of specific: nonspecific caudate and putamen uptake were consistent with the results of visual inspection. CONCLUSION: Visual assessment of [123I]-FP-CIT SPECT images is an easily applied diagnostic test which is helpful in the differential diagnosis of tremor disorders and in confirming a clinical diagnosis of a hypokinetic-rigid syndrome. PMID- 10830417 TI - Motor initiation and execution in essential tremor and Parkinson's disease. AB - Clinical differentiation of essential tremor (ET) from idiopathic Parkinson's disease (iPD) is based on the lack of akinesia and bradykinesia. Nevertheless, early tremor-predominant iPD often is difficult to distinguish from ET. Motor initiation and execution in ET, iPD, and normal control (NC) subjects were investigated. Individuals with iPD, ET and NC performed a reaction-time wrist flexion and extension task. Motor performances were similar between ET and iPD and both were different than normal control subjects. Both the patients with iPD and ET had longer reaction times and slower movement velocities than NC subjects. This may help to explain some of the difficulties in distinguishing patients with these two diseases. The similarities of motor performance suggest that while ET and iPD may be separate disease entities, they may share similar pathogenic motor mechanisms from the perspective of an integrated motor system that drives the motor cortex. PMID- 10830418 TI - Validity and test-retest reliability of a disability questionnaire for essential tremor. AB - BACKGROUND: One important outcome in clinical trials is patients' own opinions about whether the medication alleviates their symptoms and improves their ability to function. A valid and reliable method with which to assess this subjective information is important. OBJECTIVE: To determine the validity and test-retest reliability of the Columbia University Disability Questionnaire for Essential Tremor (ET). METHODS: Patients with ET underwent a 2.5-hour evaluation, including a 36-item tremor disability questionnaire, to assess the functional impact of tremor, a 26-item videotaped tremor examination rated by a neurologist, a 15-item performance-based test, and quantitative computerized tremor analysis. We determined the validity and test-retest reliability of the tremor disability questionnaire. Correlations between variables were assessed using Pearson's correlation coefficients and test-retest reliability with the weighted kappa statistic. RESULTS: Ninety-five patients with ET participated. The score on tremor disability questionnaire correlated with the neurologist's clinical ratings (r = 0.57, p <0.001) and the total score on the performance-based test (r = 0.69, p < 0.001). Correlations with quantitative computerized tremor analysis results were less robust, but each remained significant, including mean amplitude of dominant arm tremor while arms were extended (r = 0.56, p <0.001), while drawing a spiral (r = 0.42, p = 0.01), and while pouring (r = 0.34, p = 0.04). The questionnaire was readministered to 32 subjects, and the test-retest reliability was substantial (weighted kappa = 0.67). CONCLUSIONS: This Tremor Disability Questionnaire demonstrated substantial reliability, and it correlated with multiple measures of tremor severity, including a neurologist's clinical ratings, a performance-based test of function, and quantitative computerized tremor analysis results. The questionnaire would be useful in clinical trials in which it could be used as a reliable and valid tool to assess disability in ET. PMID- 10830419 TI - Delayed onset of progressive dystonia following subacute 3-nitropropionic acid treatment in Cebus apella monkeys. AB - Delayed abnormal movements can be observed in patients with acute neurologic insult after a prolonged period of apparent neurologic stability. To reproduce such a secondary neurologic manifestation in primates, the present experiment investigated whether systemic administration of subacute 3-nitropropionic acid (3NP), a mitochondrial toxin, could induce abnormal movements that were delayed and progressive over time. Four Cebus apella monkeys received systemic 3NP injections until acute neurologic signs manifested. The monkeys were regularly video-recorded and rated for abnormal movements for up to 15 weeks after the cessation of 3NP treatment. Five to 6 weeks after the 3NP treatment, monkeys displayed a significant increase in dyskinesias compared with pretreatment conditions. Over time the chorea attenuated, whereas the dystonic movements increased in intensity and severity which was characterized by a delayed decrease of peak tangential velocity. The intensity of abnormal movements and extent of affected body regions observed in each monkey were consistent with the size of basal ganglia hypersignal as documented by T2 sequence on magnetic resonance imaging. Thus, more severe motor impairments were associated with large magnetic resonance image abnormalities. This novel primate model may be particularly useful for studying the structural changes underlying delayed and progressive manifestations of abnormal movements with the ultimate goal of facilitating the evaluation of novel therapeutic strategies. PMID- 10830420 TI - Toward a primate model of L-dopa-unresponsive parkinsonism mimicking striatonigral degeneration. AB - We developed a primate model of striatonigral degeneration (SND), the neuropathology underlying levodopa-unresponsive parkinsonism associated with multiple systemic atrophy (MSA-P), by sequential systemic administration of 1 methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and 3-nitropropionic acid (3NP) in a Macaca fascicularis monkey. L-Dopa-responsive parkinsonian features emerged after MPTP injections. Subsequent chronic 3NP administration aggravated the motor symptoms and abolished the L-dopa response. In vivo magnetic resonance imaging revealed bilateral striatal lesions. Histopathologically, there was severe dopaminergic cell loss in the substantia nigra pars compacta compared with the control monkey. Furthermore, we observed circumscribed areas of severe neuronal degeneration in the motor striatum. These changes were absent in the control monkey, and they were associated with diffuse metabolic failure as demonstrated by cytochrome oxidase histochemistry. The striatal pathology predominantly involved output pre-pro-enkephalin A- and substance P-containing cells, whereas somatostatin (NADPH-diaphorase)-containing interneurons were relatively spared. Our model therefore reproduced levodopa-unresponsive parkinsonism and SND-like pathologic changes characteristic of MSA-P. The double-lesion primate model of SND may serve as a preclinical test-bed for the evaluation of novel therapeutic strategies in MSA-P. PMID- 10830421 TI - Parkinsonism, dystonia, and hemiatrophy. AB - Hemiatrophy has been reported in association with a variety of neurologic conditions, including parkinsonism. Patients with the hemiparkinson-hemiatrophy syndrome (HP-HA) have asymmetric parkinsonism with limb atrophy on the more affected side. Several authors have suggested that asymmetric brain damage early in life results in both atrophy and parkinsonism. Dopa-responsive dystonia (DRD) is a disease in which a deficiency of tetrahydrobiopterin, or, less commonly, of tyrosine hydroxylase, results in levodopa-responsive dystonia with parkinson features in children. We have recently identified four patients with DRD who had asymmetric dystonia and limb atrophy on the more affected side. Based on these patients, we suggest that a deficiency of the nigrostriatal dopamine system may, by itself, be sufficient to cause body atrophy and may underlie the limb atrophy in both DRD and HP-HA. PMID- 10830422 TI - Calcium channel agonists and dystonia in the mouse. AB - Systemic administration of the L-type calcium channel agonists +/-Bay K 8644 or FPL 64176 causes a characteristic pattern of motor dysfunction in normal C57BL/6J mice that resembles generalized dystonia. There is no associated change in the electroencephalogram, confirming that the motor disorder does not reflect epileptic seizures. However, the electromyogram reveals an increase in baseline motor unit activity with prolonged phasic discharges consistent with dystonia. The duration and severity of dystonia is dependent on the dose administered and the age of the animal at testing. The effects are transient, with the return of normal motor behavior 1-4 hours after treatment. Similar effects can be provoked by intracerebral administration of small amounts of the drugs, indicating a centrally mediated response. Dystonia can be attenuated by co-administration of dihydropyridine L-type calcium channel antagonists (nifedipine, nimodipine, and nitrendipine) but not by non-dihydropyridine antagonists (diltiazem, verapamil, and flunarizine). These results implicate abnormal function of L-type calcium channels in the expression of dystonia in this model. PMID- 10830423 TI - Rate of caudate atrophy in presymptomatic and symptomatic stages of Huntington's disease. AB - Previous research by our group demonstrated a longitudinal change in caudate volume for symptomatic subjects with Huntington's disease (HD), and suggested that volume of the caudate may be a useful outcome measure for therapeutic studies in symptomatic patients. The current study was designed to determine whether longitudinal change in caudate atrophy could be documented in presymptomatic carriers of the HD gene mutation, and to compare rate of change in these subjects with rate of change in mildly and moderately affected symptomatic patients. We measured caudate volumes on serial magnetic resonance image scans from 30 patients at three stages of HD: 10 presymptomatic; 10 with mild symptoms, as indicated by scores on the Quantified Neurological Exam (QNE) < or =35; and 10 with moderate symptoms (QNE >45). The mean interscan interval was 36 months. When analyzed separately, both symptomatic groups and the presymptomatic group demonstrated a significant change in caudate volume over time. Amount of change over time did not differ significantly among the three groups. We conclude that change in caudate volume may be a useful outcome measure for assessing treatment effectiveness in both presymptomatic and symptomatic subjects. PMID- 10830424 TI - Movement disorders in 30 patients with tuberculous meningitis. AB - BACKGROUND: Movement disorders may appear during tuberculous meningitis (TbM). OBJECTIVE: To investigate the variety of movement disorders seen in TbM and consider possible pathogenic mechanisms. DESIGN: We established two diagnostic categories for TbM: definite and probable. All patients were examined clinically and with laboratory tests, computed tomographic scan, or magnetic resonance imaging. SETTING: A university hospital in Quito, Ecuador. RESULTS: Thirty of 180 consecutively studied patients with TbM developed movement disorders. Twenty-four months after treatment was completed, we determined a prognosis for the patients. Seven patients had chorea, three dystonia, and 20 tremor. One of the patients with tremor also had myoclonus and one with dystonia had tremor. The average age of the patients with chorea was lower than that of the patients with dystonia and tremor. Two patients with chorea, one with dystonia, and three patients with tremor died. The patients with chorea and dystonia had more severe disease. We found little correlation between the type, distribution, or severity of abnormal movements and the computed tomography scan or magnetic resonance image findings. CONCLUSIONS: Tremor is the most common movement disorder seen in the course of TbM. Chorea is more frequently found in young children. Deep vascular lesions are more common among patients with movement disorders. PMID- 10830425 TI - Violent recurrent ballism associated with infections in two children with static encephalopathy. AB - A variety of cerebral insults can result in static encephalopathy with developmental delays and relatively fixed motor and cognitive deficits. We describe two boys with static encephalopathy who experienced recurrent episodes of generalized, violent ballism seemingly provoked by relatively minor infectious illnesses or surgical procedures. These episodes first began at ages 14 and 9 years, respectively. The baseline clinical states included relatively mild choreoathetosis plus cognitive impairment, as well as spasticity and/or ataxia. These episodes of ballism developed over hours, remained for weeks, and ultimately returned to baseline. Neuroleptics, anticonvulsants, and benzodiazepines were only partially beneficial; responses corresponded to the degree of sedation. Potential for self-injury or rhabdomyolysis/myoglobinuria led to the use of general anesthetics or neuromuscular blocking agents during selected episodes. Blood, urine, and cerebrospinal fluid studies, magnetic resonance imaging head scans, and electroencephalography revealed no diagnostic clues as to the precise causative factor precipitating these episodes. PMID- 10830426 TI - Posterior column ataxia and retinitis pigmentosa: a distinct clinical and genetic disorder. AB - Autosomal recessive posterior column ataxia and retinitis pigmentosa (PCARP) is a movement disorder that was genetically mapped to a disease locus (AXPC1) on chromosome 1q32-q31 in an inbred population of Dutch-German ancestry in the continental United States. We performed genetic linkage analysis and haplotype reconstruction on a different family from Spain with an identical phenotype to determine if the neurologic signs of an early-onset ataxia, retinitis pigmentosa, and a sensory neuropathy also mapped to the AXPC1 locus. The disease phenotype was linked in the candidate interval with a maximum lod score of 3.56 at a recombination fraction of 0.0 for locus D1S414. Haplotypes were discordant and suggested that the disease mutation arose independently from at least two populations. These results refine the classification of early-onset ataxia, abrogate a founder effect for this recessive disorder, and provide evidence that PCARP is a distinct, homogeneous, clinical, and genetic disorder. PMID- 10830427 TI - Clozapine replacement by quetiapine for the treatment of drug-induced psychosis in Parkinson's disease. PMID- 10830428 TI - Posttraumatic tremor and Arnold Chiari malformation: no sign of compression, but cure after surgical decompression. PMID- 10830429 TI - Lazarus' sign in brain death. PMID- 10830430 TI - Parkinsonism following electrical injury to the hand. PMID- 10830431 TI - Cloning a novel developmental regulating gene, Xotx5: its potential role in anterior formation in Xenopus laevis. AB - The vertebrate Otx gene family is related to otd, a gene contributing to head development in Drosophila. In Xenopus, Xotx1, Xotx2, and Xotx4 have already been isolated and analyzed. Here the cloning, developmental expression and functions of the additional Otx Xenopus gene, Xotx5 are reported. This latter gene shows a greater degree of homology to Xotx2 than Xotx1 and Xotx4. Xotx5 was initially expressed in Spemann's organizer and later in the anterior region. Ectopic expression of Xotx5 had similar effects to other Xotx genes in impairing trunk and tail development, and especially similar effects to Xotx2 in causing secondary cement glands. Taken together, these findings suggest that Xotx5 stimulates the formation of the anterior regions and represses the formation of posterior structures similar to Xotx2. PMID- 10830432 TI - Spatio-temporal expression of Xenopus vasa homolog, XVLG1, in oocytes and embryos: the presence of XVLG1 RNA in somatic cells as well as germline cells. AB - The expression of Xenopus vasa homolog or XVLG1 was examined in oocytes and embryos by whole-mount in situ hybridization and reverse transcription polymerase chain reaction (RT-PCR). To confirm the results in embryos, both methods were also applied to explants of germ plasm-bearing cells (GPBC) from 32-cell embryos and to those of partial embryos deprived of GPBC. By hybridization, XVLG1 ribonucleic acid (RNA) was shown to be present throughout the cytoplasm in oocytes at stages I-III, except for the mitochondrial cloud. It was barely recognizable in a portion of germline cells of embryos at specific stages, notwithstanding that XVLG1 protein was present in those cells almost throughout their life-span. A weak signal for the RNA was detectable in some of the presumptive primordial germ cells (pPGC, descendants of GPBC from the gastrula stage onward) from the late gastrula (stage 12) to the hatching tadpole stage (stage 33/34), and in some of the PGC at stages 49-50. The results for pPGC were confirmed by the hybridization of explants of GPBC at equivalent stages in control embryos. In contrast, XVLG1 RNA was detected in certain somatic cells of embryos until stage 46. These observations were supported in part by the results of RT-PCR for embryos and explants. The possible role of the product of XVLG1 was reconsidered given its presence in both germline and somatic cells. PMID- 10830433 TI - Real-time observation of transplanted 'green germ cells': proliferation and differentiation of stem cells. AB - To elucidate the mechanism of proliferation and differentiation of testicular germ cells, donor testicular germ cells labeled with enhanced green fluorescent protein (eGFP) were transplanted to recipient seminiferous tubules. The kinetics of colonization as well as of differentiation of the donor cells was followed in the same transplanted tubules (alive) under ultraviolet light. One week after transplantation, clusters of fluorescent cells were randomly spread as dots in the recipient seminiferous tubule, whereas non-homed cells flowed out from the testis to the epididymis. By 4 weeks after transplantation, green germ cells were observed with weak and moderate fluorescence along the recipient seminiferous tubule. By 8 weeks, proliferation and differentiation of the germ cells occurred, resulting in strong fluorescence in the middle part of the seminiferous tubule but in weak and moderate fluorescence at both terminals. The length of the fluorescent positive seminiferous tubule became longer. Detailed histological analyses of the recipient tubules indicated that the portions of the seminiferous tubule in weak, moderate, and strong fluorescence contained the spermatogonia, spermatogonia with spermatocytes, and all types of germ cells including spermatids, respectively. Thus, testicular stem cells colonized first as dots within 1 week, and then proliferated along the basement membrane of the seminiferous tubules followed by differentiation. PMID- 10830434 TI - Distinct expression of midkine and pleiotrophin in the spinal cord and placental tissues during early mouse development. AB - Midkine and pleiotrophin comprise a family of heparin-binding growth factors, and are expressed in overlapping tissues during the mid- to late-gestation periods of mouse development. Their distinct expression during early mouse development, as revealed by in situ hybridization, was reported. Midkine was expressed in the embryonic ectoderm from as early as embryonic day (E5.5). In the neural tube midkine was expressed specifically in the neuroepithelium, that is, in the whole area of the neural tube at E9.5, and in the ventricular zone from E10.5-13.5. At E15.5, when the neuroepithelium disappeared, midkine concomitantly became undetectable. In contrast, pleiotrophin expression started exclusively in the neural plate at E8.5, and in the lateral plate of the neural tube at E9.5. It then became restricted to a dorsal ventricular zone from E11.5-13.5, and finally to the central gray neurons at E15.5. Moreover, pleiotrophin was expressed in the ventral horns. Among placental tissues, midkine was detected in the chorion, the fetal component of the placenta, whereas pleiotrophin was found in the decidua basalis, the maternal component of the placenta. The distinct expression of midkine and pleiotrophin suggests their differential role in early development. PMID- 10830435 TI - Heteroplasmic conjugates formed by the fusion of starfish oocyte pairs with a 12 minute time difference in the maturation phase. AB - Two starfish oocytes with a 12 min time difference in the maturation phase were fused together with electric pulses to make a heteroplasmic conjugate. The starfish used were Asterina pectinifera. The emergence of the first meiotic spindle and the extrusion of the polar bodies in the conjugate were timed. Under polarization microscopy two meiotic spindles emerged with a time difference of 10 11 min, which is close to the time difference in the maturation phase between the original oocytes before fusion. In contrast, subsequent formation of the first two polar bodies occurred successively with a short time lag of 1-3 min between them. Times for the formation of both polar bodies were midway between the anticipated times for polar body formation in respective non-fused control oocytes. Thus, in one nucleus the meiotic division was delayed, while in another nucleus it was accelerated, in a single heteroplasmic conjugate. These two sets of observations indicate the presence of a certain control system that regulates progression of the cell cycle at a point during the period from the entry into metaphase through to late anaphase of meiosis I in starfish oocytes. This type of cell cycle control in starfish oocytes is obviously distinct from the currently accepted view of the cell cycle control by the spindle assembly checkpoint that monitors unattached kinetochores of mitotic chromosomes. PMID- 10830436 TI - Cyclic AMP- and calmodulin-dependent phosphorylation of 21 and 26 kda proteins in axoneme is a prerequisite for SAAF-induced motile activation in ascidian spermatozoa. AB - Sperm activating and -attracting factor (SAAF), derived from the egg of the ascidian Ciona, activates sperm motility through adenosine 3':5'-cyclic monophosphate (cAMP)-synthesis. A demembranated preparation of intact immotile sperm without SAAF was shown to require cAMP for reactivation. However, a demembranated preparation of intact motile sperm treated with SAAF did not require cAMP for reactivation, suggesting that cAMP is a prerequisite factor for SAAF-dependent activation of sperm motility. Furthermore, a cAMP-dependent protein kinase (PKA) inhibitor, H-89, was found to inhibit sperm motility. During in vivo or in vitro activation of sperm motility by SAAF or cAMP, a 26 kDa axonemal protein and 21 kDa dynein light chain were phosphorylated, respectively, suggesting the involvement of PKA-dependent phosphorylation of these proteins in sperm activation. The calmodulin antagonist, W-7, and an inhibitor of calmodulin dependent myosin light chain kinase, ML-7, also inhibited the activation of sperm motility. Inhibition was reversed by the addition of phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine. Demembranated preparations of immotile sperm in the presence of W-7 or ML-7 were reactivated by cAMP, suggesting that calmodulin participated in sperm activation and that cAMP synthesis was followed by activation of a calmodulin-dependent mechanism. PMID- 10830437 TI - Nuage constituents arising from mitochondria: is it possible? AB - An ultrastructural study of nuage-mitochondria complexes in spermatogonia of the sea urchin, Anthocidaris crassispina, was carried out. Release of mitochondrial contents into the cytoplasm was observed. The mitochondrial derivatives persisted as cristae-containing globules of friable material that subsequently contacted and integrated with nuage. The present ultrastructural findings agree with the results of other researchers who proposed that germ plasm substance probably produced by the nucleus is supplemented by the mitochondrial genome. PMID- 10830438 TI - Stages of normal tracheo-bronchial development in rat embryos: resolution of a controversy. AB - The embryonic events surrounding tracheo-esophageal separation remain controversial. The present study was undertaken to clarify early tracheo bronchial development in the rat embryo at a critical period of organogenesis. Twenty-seven timed-mated Sprague-Dawley rats were divided into nine groups of three rats. Their embryos were harvested on gestational days 11-15 at intervals of 8 h, processed and sectioned transversely. The sections were stained with hematoxylin and eosin and examined serially. The foregut is a single tube on gestational day 11. During the following 16 h, there is localized and rapid growth of the respiratory epithelium and a laterocaudal expansion to form the bronchial buds and a protuberance on the ventral wall of the foregut (future tracheal carina). From gestational days 12-12 + 8, cellular debris and apoptotic epithelial cells are specifically located in the tracheo-esophageal groove, resulting in collapse and fusion of the lateral walls of the foregut, effectively separating the trachea and esophagus. Afterwards, the epithelial proliferation dominates the process of tracheo-esophageal separation until it reaches the caudal end of the laryngeal epithelial lamina on gestational day 15. The present study shows that separation of the trachea from the esophagus involves three consecutive stages: (i) epithelial proliferation resulting in the formation of bronchial buds and the tracheal carina; (ii) epithelial apoptosis leading to separation of the trachea and esophagus; and (iii) epithelial proliferation to complete the separation process. PMID- 10830439 TI - Relationship between ATP level and respiratory rate in sea urchin embryos. AB - In sea urchin embryos, the rate of respiration, as a result of electron transport through the mitochondrial respiratory chain, was enhanced after hatching without any change in the intrinsic capacity of electron transport in mitochondria. The increase in respiratory rate after hatching was accompanied by an evident decrease in intracellular adenosine triphosphate (ATP) concentration without any change in intracellular levels of adenosine diphosphate (ADP) and adenosine monophosphate (AMP). Adenosine triphosphate is proposed to fortify acceptor control of respiration at high concentrations and to reduce the respiratory rate even in the presence of ADP, the acceptor. The relationships between the respiratory rate and intracellular ATP concentration in embryos were the same as those in mitochondria isolated from embryos, obtained in the presence of ADP at the same concentration as in the embryos. Probably, the respiratory rate is enhanced after hatching because of the decrease in the level of ATP. In embryos kept in a medium containing adenosine, intracellular ATP concentration increased especially after hatching, without any change in the ADP level, and the respiratory rate after hatching was made as low as the rate expected, based on the relationships obtained on isolated mitochondria. The respiratory rate in embryos probably depends on intracellular ATP concentration, irrespective of the developmental stage in early development. PMID- 10830440 TI - Cloning of a cDNA for Xenopus prolactin receptor and its metamorphic expression profile. AB - A pituitary hormone, prolactin (PRL) shows various effects on cellular metabolism in amphibians, such as stimulation of larval tissue growth and inhibition of metamorphic changes. All these effects are mediated by its cell surface receptor. However, lack of information on PRL receptor (PRL-R) gene expression has made the physiological importance of the PRL/PRL-R system obscure in amphibian metamorphosis. Hence, a Xenopus PRL-R cDNA was cloned, its structure was characterized, and specific binding of PRL to Xenopus PRL-R expressed in COS-7 cells was confirmed. In adult tissues, high level expression was found in the lung, heart, brain, thymus and skin, and low level in the oviduct, kidney and spinal cord. The developmental expression pattern showed that PRL-R messenger ribonucleic acid (mRNA) was expressed in the brain and tail from premetamorphosis and the level increased toward late metamorphosis, suggesting that PRL may inhibit the metamorphic changes in those organs. The level of brain PRL-R mRNA reached a peak just at the start of the metamorphic climax stages and then decreased, whereas in the tail, mRNA expression peaked at late metamorphosis. In the kidney, mRNA expression increased and reached a maximum level at the end of metamorphosis. The results obtained were discussed in relation to metamorphosis. PMID- 10830441 TI - In vitro organogenesis of pancreas in Xenopus laevis dorsal lips treated with retinoic acid. AB - Dorsal lips of Xenopus laevis may differentiate into pancreas after treatment with retinoic acid in vitro. The dorsal lip region is fated to be dorsal mesoderm and anterior endoderm. Dorsal lip cells isolated from stage 10 early gastrula differentiate into tissues such as notochord, muscle and pharynx. However, in the present study, dorsal lips treated with 10(-4) M retinoic acid for 3 h differentiated into pancreas-like structures accompanied by notochord and thick endodermal epithelium. Sections of the explants showed that some cells gathered and formed an acinus-like structure as observed under microscopes. In addition to the morphological changes, expressions of the pancreas-specific molecular markers, XIHbox8 and insulin, were induced in retinoic acid-treated dorsal lip explants. Therefore, it is suggested that retinoic acid may induce the dorsal lip cells to differentiate into a functional pancreas. However, continuous treatment with retinoic acid did not induce pancreas differentiation at any concentration. Dorsal lips treated with retinoic acid within 5 h after isolation differentiated into pancreas-like cells, while those treated after 15 h or more did not. The present study provided a suitable test system for analyzing pancreas differentiation in early vertebrate development. PMID- 10830442 TI - Beneficial effect of nicotinamide on the proportion of insulin cells in developing chick pancreas. AB - Previous studies have suggested that nicotinamide increases the number of insulin cells both in vivo and in vitro. However, the question remains as to whether there is in fact an increase and whether this increase is caused by the proliferation of progenitor cells, or by replication of existing insulin cells. In order to investigate this, the endodermal component of dorsal pancreatic buds of 5-day-old chick embryos was cultured on Matrigel in a serum-free medium (Ham's F12-ITS) to which nicotinamide, at a concentration of 5 and 10 mM, respectively, was added. Control explants were cultured in Ham's F12-ITS medium without nicotinamide. After 7 days in culture the buds were incubated with bromodeoxyuridine (BrdU) and then processed for immunocytochemistry. Localization of insulin, BrdU and glucagon was carried out on adjacent serial sections. The proportion of insulin cells was 6.76, 11.32 and 16.86% in control, 5 and 10 mM nicotinamide-treated explants, respectively. Hence adding nicotinamide to the culture medium induced a 1.7- and 2.5-fold increase in the proportion of insulin cells when compared to the controls. These proportions were significantly different from that of control explants (P < 0.05). However, a very small number of insulin cells were found to be proliferating, suggesting that the increase in the proportion of insulin cells had resulted from stimulation of progenitor cells and not proliferation of existing insulin cells. PMID- 10830443 TI - Children as victims of war and conflict: the challenged rights of the child. PMID- 10830444 TI - Protection of the young child's brain: personal observations and thoughts in postwar stress syndrome and in natural catastrophes. The Nils Rosen von Rosenstein Lecture at Uppsala University, 6 May 1999. AB - This paper calls attention to the plight of children exposed to various traumas associated with wars and other natural disasters around the world. Little systematic attention has been paid to the mental health needs of child survivors of armed conflict. Our observations since 1968 and experiences around the world suggest that there are significant differences in the degree to which children have been prepared in anticipation of exposure to violence and in interventions applied after their exposure. It is the author's impression that prevention and intervention can be quite powerful when employed in these situations. PMID- 10830445 TI - Childhood cancer--mainly curable so where next? AB - More than 70% of childhood cancer is now curable with best modern therapy. The treatment is expensive but in terms of cost per life year saved, USD 1750, compares very favourably with other major health interventions. The rate of improvement in survival is slowing down. New, "designer", treatments are needed and, better still, prevention. The causes of childhood cancer are beginning to emerge. The origin for many is probably in utero and may be initiated by dietary and other environmental exposures perhaps in susceptible individuals. However, one of the great challenges for the future must be to extend the benefits of modern treatment to the 80% of the world's children who currently have little or no access to it in economically disadvantaged and emerging nations. The International Paediatric Oncology Society (SIOP) is leading the way in bringing hope for children with cancer worldwide. In India, with the support of the WHO, there is a "train the trainers" programme. In Africa, pilot studies of cost effective treatments for Burkitt's lymphoma are producing gratifying results in Malawi and there are several examples of twinning programmes between major centres in developed and less well-developed countries. CONCLUSIONS: The future for children with cancer is bright. Most are curable and prevention may be just over the horizon. PMID- 10830446 TI - Multi-system approach to the assessment of successful feeding. PMID- 10830447 TI - Direct and indirect protection from Hib disease: may immunization of a target group lead to eradication of the disease in a population? PMID- 10830448 TI - Salt intake in early life and cardiovascular risk. PMID- 10830449 TI - Fatty acid composition of serum cholesterol esters as a reflector of low saturated-fat, low-cholesterol diet in young children: the STRIP project. The Special Turku coronary Risk factor Intervention Project. AB - STRIP (the Special Turku coronary Risk factor Intervention Project) is an ongoing intervention trial which aims at a permanent reduction in the intake of saturated fat and cholesterol starting in childhood. A total of 75 intervention and 63 control children was studied consecutively at the ages of 7 and 13 mo, and 2, 3 and 5 y to evaluate the influence of such intervention on serum cholesterol ester (CE) fatty acid composition, a widely used biomarker of fatty acid intake. Analysis of 4-d food records showed that total intake of fat and of saturated fat increased with age in both groups of children but was constantly lower in intervention than in control children, e.g. at the age of 5 y the mean intakes of total fat and of saturated fatty acids were 31.1 E% and 33.9 E% and 12.1 E% and 14.6 E% in intervention and control children, respectively (p = 0.009 and 0.0001, respectively). Serum CE fatty acid compositions did not differ between the 2 groups at any age; the mean proportion of CE linoleic acid was 52.4% and 52.0% in 5-y-old intervention and control children, respectively. Correlation analysis showed, however, that the percentage of linoleic acid and of polyunsaturated fatty acids in CE reflected well the respective dietary intakes (r = 0.36; p = 0.0001 for both coefficients). In conclusion, CE fatty acid composition did not differ between the intervention and control groups, whereas CE linoleic and total polyunsaturated fatty acids reflected well the differences in their intakes at the individual level. PMID- 10830450 TI - Sodium intake of 1 to 5-year-old children: the STRIP project. The Special Turku Coronary Risk Factor Intervention Project. AB - The aim of this study is to examine sodium intake and dietary sodium sources of 1 5-y-old children in a prospective, randomized long-term coronary heart disease prevention trial, focused on dietary fat modification. Counselling included no advice about reducing salt in the children's diets. Food consumption of 100 intervention children and 100 control children was recorded for 3 consecutive days at the age of 13 mo and for 4 consecutive days at the ages of 3 and 5 y. Sodium intakes were calculated using the Micro Nutrica program. Children's mean daily sodium (NaCl) consumption (intervention and control children combined) was 1600+/-527 mg (4.0+/-1.3 g), 1900+/-504 mg (4.8+/-1.3 g) and 2200+/-531 mg (5.5+/ 1.3 g) at the ages of 13 mo and 3 and 5 y, respectively. The intervention children consumed as much or slightly more sodium than the control children at all ages studied. Half the sodium consumption was derived from added salt in commercially prepared or homemade foods. Milk, meat products, bread and cereals were other important sodium sources. In conclusion, nutrition counselling in the Special Turku Coronary Risk Factor Intervention Project (STRIP) trial, with its main focus on the quality of fat in child nutrition, has had minimal influence on children's sodium intake. To avoid excessive sodium intake in children, dietary counselling should include information about salt use, and food manufacturers should be encouraged to provide more low-sodium products. PMID- 10830451 TI - Hypo-osmolar oral rehydration salts solution in dehydrating persistent diarrhoea in children: double-blind, randomized, controlled clinical trial. AB - A double-blind, randomized, controlled trial was conducted to compare the clinical efficacy of hypo-osmolar oral rehydration salts (ORS) solution (224 mmol/L) and standard ORS solution (311 mmol/L) in children with persistent diarrhoea who were prone to develop dehydration. Initially, 95 children aged between 3 and 24 mo were included in the study for overnight observation. Of these, 70 children who passed stool more than 2 g/kg/h were finally enrolled in the study and were randomly assigned either standard ORS or hypo-osmolar ORS. After decoding the identity of ORS, it was observed that 37 children were in the standard ORS group and 33 in the hypo-osmolar ORS group. Clinical parameters and microbiological findings of stool samples were comparable in the two groups at the time of enrolment. Total stool output (2.5+/-1.1 vs 3.2+/-1.6 kg; p = 0.04), duration of diarrhoea (114.8+/-38.3 vs 145.4+/-40.0 h; p = 0.002), total intake of ORS (5.4+/-1.6 vs 7.8+/-1.8 l; p = 0.002) and total fluid intake (7.9+/-2.6 vs 10.0+/-4.1 l, p = 0.01) were significantly less in the hypo-osmolar ORS group compared to the standard ORS group. However, the percentage of weight gain on recovery in the hypo-osmolar group was less compared to that of the standard ORS group, though the difference was statistically insignificant. Thirty-five (95%) children in the standard ORS and 33 (100%) children in the hypo-osmolar group recovered within 10 d of initiation of therapy and modified dietary management. CONCLUSION: Our findings suggest that hypo-osmolar ORS has beneficial effects on the clinical course of dehydrating persistent diarrhoea. PMID- 10830452 TI - Evolving experience in the management of empyema thoracis. AB - The optimal management of paediatric empyema thoracis remains controversial. The objective of the study was to analyse evolving experience in clinical presentation, management, outcome and factors contributing to adverse morbidity in thoracic empyema. Forty-seven patients presenting to a paediatric surgical centre were studied in three consecutive 6-y periods during 1980-97 to compare any change in the pattern of disease influencing diagnosis and management. Patients were categorized into two treatment groups: (i) conservative management (antibiotics and/or tube thoracostomy), (ii) thoracotomy. The median duration of illness prior to hospital admission was 10 d (range 1-42 d). Ultrasound was increasingly utilized in the diagnosis and staging of empyema and played an important role in directing definitive management. The presence of loculated pleural fluid determined the need for thoracotomy. Sixteen of 20 patients (80%) who were initially treated with thoracocentesis or tube thoracostomy eventually needed thoracotomy. There was a positive shift in management towards early thoracotomy resulting in prompt symptomatic recovery. Significant complications were noted in seven children who had delayed thoracotomy. These included recurrent empyema with lung abscess (n = 2), scoliosis (n = 2), restrictive lung disease (n = 1), bronchopleural fistula (n = 1) and sympathetic pericardial effusion (n = 1). An unfavourable experience with delayed thoracotomy during the study period has led us to adopt a more aggressive early operative approach to empyema thoracis. The decision to undertake thoracotomy has been influenced by the ultrasound findings of organized loculated pleural fluid. Delayed surgery was associated with adverse outcome. Whilst fibrinolytics and thoracoscopy may provide attractive options for early empyema, thoracotomy can hasten patient recovery regardless of the stage of disease. Prospective randomized trials are required to assess the ideal therapy for childhood empyema. PMID- 10830453 TI - Duration of secretory IgM and IgA antibodies to respiratory syncytial virus in a community study in Guinea-Bissau. AB - Respiratory syncytial virus (RSV) is probably the single major cause of lower respiratory infection (LRI) among infants worldwide. Its relative importance may be underestimated, as the diagnosis is based on antigen detection and antigen may only be detectable in the early phase of infection. We have therefore assessed the duration of secretory IgM and IgA antibody responses and whether assays for these antibodies can be used to improve the diagnosing of RSV-associated infections. During two RSV epidemics in Guinea-Bissau, 32 RSV antigen-positive children with LRI were followed with sequential nasopharyngeal suction on days 7, 14, 30, 60 and 120 in the first epidemic and every fortnight for 6 mo after the second epidemic to measure the duration of secretory IgM and IgA responses. Nearly all of the children had an IgM response during the first month after infection. The response ratio was highest on days 7 and 14, being 84% and 71%, respectively. After 30 d the IgM response decreased rapidly. Among 27 age- and sex-matched controls, only 1 child was positive for IgM. During the second epidemic, when the children were followed more intensively, half of the children were IgM-positive after the acute phase of infection. A secondary response may be more likely in children with low IgM responses in the acute phase (RR = 2.08 (95% confidence interval (CI) 0.92-4.70)). The IgA response was highest on days 28 and 42 after antigen detection, 72% having a detectable IgA response within the first 1.5 mo. Among 27 controls, only 2 were IgA-positive (7%). In the second epidemic with more intensive follow-up, 62% (8/13) of the IgA-positive children had a response that lasted 10 wk. Of the children with no persistent IgA response, half (5/10) had a subsequent IgA-positive response after the first 42 d. All of these children had a simultaneous IgM-positive response. When 29 of the children were tested after an epidemic when they were 1-3-y-old, >80% again had high IgM (24/29, 82%) and IgA (28/29, 94%) levels. Among samples collected over a 1-y period from infants with LRI in a community morbidity surveillance conducted at the local health centre and via paediatric outpatient consultation, 17% (110/659) were antigen-positive, 26% (171/659) IgM-positive and 38% (248/659) either antigen- or IgM-positive. IgM responses are short-lived among infants and may therefore be used as an indication of recent RSV infection among children with LRI. Using both antigen and IgM detection may significantly improve our detection of RSV infections. PMID- 10830455 TI - Vulvar symptoms in paediatric and adolescent patients. AB - Vulvovaginal symptoms in children and young adolescents are not yet very well understood, nor is the actual incidence known. This study evaluates the character and possible infectious aetiology of vulvar symptoms of females aged up to 16 y. The signs, symptoms and bacteriological findings of 68 consecutive cases were studied. The study was conducted in the University Hospital of Tampere at a special gynaecological consultation clinic for children and adolescents. Sixty eight patients were included in the study: 48 girls (71%) were prepubertal, at Tanner stage M1P1, 26 patients were 2-4-y-old and 15 were 5-7-y-old. The duration of symptoms was known for 46 patients: 41% had had symptoms for >1 mo and 20% for >6 mo. Forty-eight patients had abnormal clinical findings on examination and 16 (33%) of them had an infectious aetiology. Streptococcus pyogenes infection was identified in 11 (16%) patients, all of whom had symptoms. Candida was identified in 6 (9%) patients. No infectious aetiology was found among 26 patients who had symptoms and abnormal clinical findings. Vulvovaginal symptoms during childhood are more common among younger children (<7 y). In 67% of patients no infectious aetiology could be found. Samples for microbiological culture should be taken from symptomatic patients and symptomatic areas. Cultures of Candida and bacteria are necessary but usually sufficient. If a microbiological aetiology is established, treatment can be assigned accordingly. Patients with vulvar symptoms and findings but with an unclear aetiology need support and advice on proper hygiene and can intermittently use mild corticosteroids locally. PMID- 10830454 TI - Paediatric infective endocarditis: 19-year experience at a tertiary care hospital in a developing country. AB - A retrospective study was undertaken to study children who presented with infective endocarditis (IE) to a university teaching hospital in Beirut, Lebanon, between January 1977 and May 1995. Of 41 patients with IE (24F, 17M), 28 (68%) were diagnosed between 1977 and 1985. Patients' ages ranged from 3 to 18 y (mean age 11.3+/-2.8 y), and 13 patients were <10 y of age. Clinical presentations included: fever (in 88%), heart failure (in 39%), neurologic findings (in 20%) and embolic phenomena (in 22%). Nineteen patients (46%) had underlying congenital heart disease (CHD) with tetralogy of Fallot and pulmonary stenosis being the most common. Sixteen patients (39%) had underlying rheumatic heart disease (RHD). A total of 5 children (12%) with normal cardiac anatomy had IE. One had underlying acquired viral myocarditis with mitral insufficiency. Echocardiography showed vegetations in 60%. Blood cultures were positive in 31 patients (76%). IE occurred in three patients following cardiac surgery. In one patient it occurred within 2 mo of surgery and in the other two it occurred within 6 mo. Streptococcus viridans and Staphylococcus aureus were the two most commonly isolated bacteria. Overall mortality rate was 29% (not statistically significant between patients presenting between 1977-1985 and 1986-1995; p = 0.17). There was no statistically significant difference in mortality among the groups (five in the group with CHD, six with RHD and one with structurally normal heart). This study demonstrates that RHD is an important underlying cause of IE in children in our community. This finding is similar to those in other developing countries and different from those in developed countries. Distribution of pathogens and CHD in our study is comparable to some reports in the literature, except for the higher proportion of patients with underlying pulmonary stenosis. Bacterial endocarditis prophylaxis should be emphasized in patients with RHD or pulmonary stenosis. PMID- 10830456 TI - Longitudinal assessment of cardiac status by echocardiographic evaluation of left ventricular diastolic function in thalassaemic children. AB - Cardiac disease is the primary cause of death in patients affected by thalassaemia major. In most cases diastolic dysfunction precedes the onset of systolic impairment at a time when appropriate therapy can prevent progression of cardiac damage. We have assessed the pattern of left ventricular filling by Doppler echocardiography in six transfusion-dependent thalassaemic patients. They were re-evaluated 5 y after the first observation, after additional blood transfusions and consequent iron load. We registered a significantly impaired relaxation pattern in all of them. A state of cardiac anoxia has been hypothesized. PMID- 10830457 TI - Simultaneous stimulation of growth hormone, adrenocorticotropin and cortisol with L-dopa/L-carbidopa and propranolol in children of short stature. AB - In 59 otherwise healthy children of short stature, the simultaneous response of growth hormone, cortisol and plasma adrenocorticotropin (ACTH) to L-dopa/L carbidopa and propranolol at 45 and 90 min after administration were investigated. A growth hormone response of 10 microg/l or higher was considered positive. The definition of a positive cortisol response included either a hormone increase of at least 193 nmol/l or a peak hormone level of at least 497 nmol/l. The ACTH increase had to be fourfold above 11 pmol/l to be considered positive. In the 59 investigated children, the median basal growth hormone levels increased from 1.35 microg/l to 18.05 microg/l and 10.15 microg/l at 45 and 90 min after stimulation (p < 0.05). The median cortisol levels rose from 242 nmol/l to 439 nmol/l and 612 pmol/l, and the corresponding median ACTH levels from 2.94 pmol/l to 9.63 pmol/l and 11.13 pmol/l at 45 and 90 min after stimulation. Significant positive hormone response rates were 88.1% for growth hormone, 88.1% for cortisol and 69% for ACTH. These results could be attributed to the enhanced stimulating effect of the additional administration of L-carbidopa and propranolol. We conclude that the administration of L-dopa/L-carbidopa and propranolol is useful for the simultaneous evaluation of growth hormone, cortisol and ACTH secretion in children of short stature. PMID- 10830458 TI - Heart risk associated with weight loss in anorexia nervosa and eating disorders: electrocardiographic changes during the early phase of refeeding. AB - Refeeding syndromes with electrolyte aberrations, heart failure and arrhythmias may complicate the nutritional rehabilitation of emaciated patients with eating disorders. Therefore, electrocardiographic (ECG) changes and changes in serum electrolyte concentrations following refeeding were studied in 37 admissions of 32 teenage girls with eating disorders. On admission they were all on a weight losing course and weighed 37.0+/-8.0 kg (mean +/- SD) following a weight loss of 14.2+/-7.2 kg. On ECG recordings there was a prolongation of the QT interval and an increased QT dispersion. Serum concentrations of sodium, potassium and magnesium were with few exceptions normal. Serum concentrations of creatinine were high in relation to the low body weight, indicating protein catabolism. The first 2 wk of refeeding resulted in a weight gain of 1.7+/-0.2 kg without signs of refeeding syndromes or electrolyte aberrations. QT prolongation and dispersion normalized within the 3 d of refeeding. It is concluded that oral refeeding of patients with eating disorders and weight loss can be performed efficiently and without causing refeeding syndromes. QT pathology, a consequence of acute starvation and a risk factor for cardiac arrhythmias, normalizes within days. In view of the need to balance adequate refeeding and reduction of QT pathology against the risks of refeeding syndromes the start of refeeding of severely emaciated patients is best performed in a hospital setting where monitoring of ECG and serum electrolytes is possible. PMID- 10830459 TI - Oral feeding in premature infants: advantage of a self-paced milk flow. AB - An earlier study demonstrated that oral feeding of premature infants (<30 wk gestation) was enhanced when milk was delivered through a self-paced flow system. The aims of this study were to identify the principle(s) by which this occurred and to develop a practical method to implement the self-paced system in neonatal nurseries. Feeding performance, measured by overall transfer, duration of oral feedings, efficiency, and percentage of successful feedings, was assessed at three time periods, when infants were taking 1-2, 3-5, and 6-8 oral feedings/day. At each time period, infants were fed, sequentially and in a random order, with a self-paced system, a standard bottle, and a test bottle, the shape of which allowed the elimination of the internal hydrostatic pressure. In a second study, infants were similarly fed with the self-paced system and a vacuum-free bottle which eliminated both hydrostatic pressure and vacuum within the bottle. The duration of oral feedings, efficiency, and percentage of successful feedings were improved with the self-paced system as compared to the standard and test bottles. The results were similar in the comparison between the self-paced system and the vacuum-free bottle. Elimination of the vacuum build-up naturally occurring in bottles enhances the feeding performance of infants born <30 wk gestation as they are transitioned from tube to oral feeding. The vacuum-free bottle is a tool which caretakers can readily use in neonatal nurseries. PMID- 10830460 TI - Pilot study of nebulized surfactant therapy for neonatal respiratory distress syndrome. AB - Thirty-four spontaneously breathing newborns with respiratory distress syndrome (RDS) requiring nasal continuous positive airway pressure (CPAP) and an arterial to-alveolar oxygen tension ratio (a/A PO2) of 0.15-0.22 were randomized to treatment with nebulized surfactant (Curosurf) or to serve as controls. All children were first supported by nasal CPAP according to normal clinical routines. Surfactant was administered using a modified Aiolos nebulizer, and a total of 480 mg was aerosolized in each case. The control group received no nebulized material, but had the same CPAP support. Acid-base status and a/A PO2 were determined at regular intervals before, during and after surfactant administration. Both groups included in the study were similar with regard to gestational age, birthweight, steroids given before birth, sex and Apgar scores as well as a/A PO2 when entering the study. There were no significant differences between the groups in a/A PO2 1-12 h after randomization, number of infants needing mechanical ventilation, time on ventilator or CPAP. Two children in the treated group developed bronchopulmonary dysplasia. No side effects of the surfactant therapy were noted. No beneficial effects of aerosolized surfactant were demonstrated in our trial, contrary to data from animal experiments. This finding probably reflects differences in administration techniques. Our findings do not justify large clinical trials with the same protocol. Further work is needed to optimize delivery of aerosolized surfactant to the neonatal lung in clinical practice. PMID- 10830461 TI - Changes in prevalence and severity of asthma among schoolchildren in a Swedish district between 1985 and 1995. AB - The same questionnaire and study design was used in two surveys of asthma among all the children attending the 9-y compulsory school in Sundsvall in 1985 (n = 10527) and 1995 (n = 9165). A detailed questionnaire was distributed by post to the parents of all children who had answered in the affirmative to a simple screening question on asthmatic symptoms at the beginning of the autumn term. The questionnaire contained detailed questions on symptoms and asthma management. Our findings indicated a moderate increase in reported asthma-like symptoms and physician-diagnosed asthma between 1985 and 1995. The severity of symptoms was unchanged, despite a large community-based asthma campaign and a tenfold increase in the number of children receiving inhaled steroids. A validation analysis included an interview by a physician, a skin prick test, determination of specific IgE antibodies and spirometry. The oral interviews suggested that undertreatment was common. Many children had adequate medication at home, but this medication was not used properly. Finally, all 13-14-y-old children also replied to written and video questionnaires from the International Study of Asthma and Allergies in Childhood (ISAAC). It is likely that differences in study design explained the much higher prevalence of wheezing in this part of the study. PMID- 10830462 TI - Invasive disease due to Haemophilus influenzae type b during the first six years of general vaccination of Swedish children. AB - Since 1992-93 vaccination against Haemophilus influenzae type b (Hib) has been included in the general Swedish childhood vaccination programme. The aim of the present study is to describe the epidemiology, identify and describe vaccine failures and calculate vaccine effectiveness during the first 6 y after introduction of vaccination against Hib. Laboratory reports of blood and cerebrospinal isolates to the Swedish Institute for Infectious Disease Control were used as the source for identifying the patients. Additional information was subsequently obtained from physicians and parents of children who had developed the disease during the study period. Vaccine failures were identified and vaccine effectiveness calculated. During the study period, 152 cases of invasive H. influenzae were identified in the age group 0-14 y. During the 6-y period, 6 true vaccine failures, 6 apparent vaccine failures and 1 possible vaccine failure were found in nearly two million vaccinated child-years. The effectiveness of the Hib vaccination in the birth cohort of children 1993 to 1997 in Sweden was calculated to be 96.1% (95% confidence interval 94.2-97.5). The study supports earlier studies from several countries that conjugated Hib vaccination introduced in general childhood vaccination programs is effective and substantially decreases suffering from invasive Hib diseases. PMID- 10830463 TI - Prepulse inhibition of startle, intelligence and familial primary nocturnal enuresis. AB - Previous studies have shown a significant reduction of prepulse inhibition of startle in boys with primary nocturnal enuresis. Those enuretic boys who had higher IQs showed less prepulse inhibition. This study evaluates the association of prepulse inhibition and IQ in primary nocturnal enuresis in respect to family history of primary nocturnal enuresis. Prepulse inhibition of startle was studied in 83 boys with primary nocturnal enuresis and 57 non-enuretic boys using an interval of 120 ms between the onset of a 75 dB 1000 Hz tone and a 104 dB noise burst. Of the boys with primary nocturnal enuresis, 56 had a family history of primary nocturnal enuresis and 27 had no family history (no first-degree relative). Of the 57 non-enuretic boys, 42 also had no family history (no first degree relative) of primary nocturnal enuresis, while 15 did have a positive family history. Associations between prepulse inhibition and IQ scores were compared among these four groups. Strong and significant associations between prepulse inhibition deficit and higher IQ scores in the enuretic group with familial primary nocturnal enuresis were unique in comparison to the other groups. CONCLUSIONS: The strong heritabilities of primary nocturnal enuresis, intelligence and prepulse inhibition suggest genetic mediation of the association of prepulse inhibition with intelligence in familial primary nocturnal enuresis. PMID- 10830464 TI - Cost analysis of the treatment of acute childhood lymphocytic leukaemia according to Nordic protocols. AB - Some attempts have been made to reduce the costs incurred in the therapy of leukaemia, but no studies are available regarding costs of the entire treatment in children with acute lymphocytic leukaemia (ALL). We analysed all the direct costs of treatment of 11 children with ALL diagnosed and treated in Kuopio University Hospital. The follow-up continued from diagnosis until the end of treatment for each patient. Patient treatment on the ward lasted for 84-210 d and in the outpatient clinic for 24-66 d, depending on the risk group. From 11-54 of the inpatient days were required for the treatment of infections. Total mean cost of the entire treatment was US $103250 (US $55196-166039) per patient, 53% of which were basic hospital costs and 47% patient-specific costs. Laboratory tests and radiology accounted for 18% of all direct costs and cytostatic drugs for 13%, but blood products accounted for only 4% of the total. Infections were the most important extra cause of costs, accounting for 18% of the mean total costs per patient. The complete treatment of a child with ALL came to a total of US $103250. However, since 80% of children with ALL are long-term survivors, the cost must be regarded as a good investment. PMID- 10830465 TI - Syndrome of inappropriate secretion of antidiuretic hormone complicating neonatal diazepam withdrawal. PMID- 10830466 TI - Neurological manifestations of deep sinus infection. PMID- 10830467 TI - Very-long-chain acyl-coenzyme A dehydrogenase (VLCAD) deficiency: monitoring of treatment by carnitine/acylcarnitine analysis in blood spots. PMID- 10830468 TI - Myopathy associated with rickets and celiac disease. PMID- 10830469 TI - An unusual case of refractory hypoglycaemia in a neonate receiving total parenteral nutrition. PMID- 10830471 TI - Infective endocarditis in children--report of 20 cases from a national survey and comparison with adult endocarditis. PMID- 10830470 TI - Therapeutic strategy in nocturnal enuresis. PMID- 10830472 TI - Gender and disease in Japan. PMID- 10830473 TI - Altered beta-adrenoceptor-mediated responses in the gastric smooth muscle of hypertensive rats. AB - Effects of isoproterenol on contraction and membrane potential of gastric smooth muscle were studied in stroke prone spontaneously hypertensive rats (SHRSP) and normotensive Wistar Kyoto rats (WKY). Circular muscle preparation from the gastric fundus developed tonic contraction by re administration of Ca2+ to a nominally Ca2+-free solution. The contraction was inhibited by nifedipine or nicardipine. Isoproterenol induced relaxation when it was applied to the Ca2+ induced contraction. The amplitude of isoproterenol induced relaxation was concentration-dependent. Propranolol 10(-6) M abolished the relaxation induced by isoproterenol 10(-7) M. In the preparation from SHRSP, the amplitude of isoproterenol induced relaxation was smaller than that from WKY between 3 x 10( 9) and 10(-7) M. Forskolin, an adenylate cyclase activator, induced concentration dependent relaxation. There was no difference in the relaxation induced by forskolin between preparations from WKY and SHRSP. Dibutilyl cyclic AMP, a membrane permeable analogue of cyclic AMP, also induced similar relaxation in preparations from WKY and SHRSP. Resting membrane potential of smooth muscle cell was not different between preparations from WKY and SHRSP. Isoproterenol hyperpolarized the membrane concentration-dependently. Isoproterenol-induced hyperpolarization in the preparation from SHRSP was smaller than that from WKY between 10(-8) and 10(-6) M. When the membrane was depolarized by Tyrode's solution containing 40 mM K+, isoproterenol-induced hyperpolarization was almost abolished. In this condition, the isoproterenol-induced relaxation was inhibited partly, however, there was no difference in the amplitude of relaxation between preparations from WKY and SHRSP. Therefore, isoproterenol-induced hyperpolarization contributed at least partly to the relaxation. Forskolin hyperpolarized the membrane by the same amplitude in the preparations from WKY and SHRSP. These results indicate that a decrease in hyperpolarization may contribute to the decreased relaxation by isoproterenol in the preparation from SHRSP. PMID- 10830474 TI - The effects of beta-adrenoceptor agonists on KCl-induced rhythmic contraction in the ureter of guinea pig. AB - In the present study, we tried to determine what effects were induced by beta adrenoceptor agonists on 40 mM KCl-induced rhythmic contraction and to clarify which beta-adrenoceptor subtypes are involved in the regulation of ureter motility in the guinea pig by using in vitro functional analysis. 40 mM KCl induced rhythmic contraction was abolished by papaverine (10(-6) M), nicardipine (10(-5) M) and cromakalim (10(-5) M), but was not influenced by atropine (10(-6) M). Isoprenaline decreased the amplitude, and changed the pattern of 40 mM KCl induced rhythmic contraction in concentration-dependent manner. These results suggest the possibility that the stimulation of beta-adrenoceptors may regulate the ureteral peristalsis. Salbutamol (selective beta2-AR agonist) and CGP12177 (beta(1,2)-AR antagonist and beta3-AR partial agonist) were also effective in decreasing the amplitude and changing the pattern of the rhythmic contraction. The pD2 values of agonists were 7.57 (isoprenaline), 5.80 (CGP12177) and 7.63 (salbutamol), respectively. The concentration-response curves of isoprenaline and salbutamol were rightward shifted by the presence of propranolol, and the apparent pA2 values for propranolol against isoprenaline and salbutamol were 7.12 and 6.29, respectively. These results suggest that inhibition for 40 mM KCl induced rhythmic contraction of the ureter by isoprenaline and salbutamol mediated mainly via atypical beta-adrenoceptor subtype. PMID- 10830475 TI - Function and distribution of beta3-adrenoceptors in rat, rabbit and human urinary bladder and external urethral sphincter. AB - 1. Activation of beta-adrenoceptors causes relaxation of the urinary bladder and contraction of the external urethral sphincter, which consists of fast contracting skeletal muscles. A beta2-adrenoceptor agonist, clenbuterol, recently has been developed as a therapeutic drug for the treatment of urinary incontinence, however beta2-adrenoceptor agonists have undesirable effects on cardiac and striated muscle function. 2. In this study, we compared the effects of the beta2-adrenoceptor agonist, clenbuterol and of a novel beta3-adrenoceptor agonist, GS332, on urinary bladder and external urethral sphincter function in rat, rabbit and human. We also determined the distribution of beta3-adrenoceptors in human urinary bladder and external urethral sphincter, using radioligand binding techniques. 3. Clenbuterol induced marked relaxations in rat, rabbit and human urinary bladder smooth muscles and also induced marked contractions in rat periurethral striated muscles (external urethral sphincter), while GS332 induced marked relaxations in rat and human, but not in rabbit, urinary bladder smooth muscles and induced small contractions in rat periurethral striated muscles. 4. The radioligand binding studies showed presence of beta2- and beta3-adrenoceptors in human urinary bladder, external urethral sphincter and abdominal rectus muscles. The affinities of GS332 were the highest in urinary bladder and the lowest in the skeletal (abdominal rectus) muscles, while the affinities of clenbuterol were similar in urinary bladder, external urethral sphincter and the skeletal (abdominal rectus) muscles. 5. These results suggest that GS332 could, similarly clenbuterol, have a role in the treatment of urinary frequency and urinary incontinence. PMID- 10830476 TI - Effect of chronic treatment with perindopril on endothelium-dependent relaxation of aorta and carotid artery in SHRSP. AB - Endothelium-dependent relaxation of aorta and carotid artery from stroke-prone spontaneously hypertensive rats (SHRSP) and the effect of chronic treatment of SHRSP with perindopril, an angiotensin converting enzyme inhibitor, on endothelium-dependent relaxation were studied. Endothelium-dependent relaxation was induced by acetylcholine (ACh) in preparations of SHRSP and normotensive Wistar Kyoto rats (WKY) precontracted with noradrenaline. The ACh-induced relaxation in both preparations was abolished by L-nitroarginine. The ACh-induced relaxation was impaired in preparations from SHRSP and contraction was observed at high concentrations of ACh. In the presence of indomethacin, impairment of endothelium-dependent relaxation in SHRSP was minimized and the contraction was inhibited. The relaxation with sodium nitroprusside did not differ between the preparations from WKY and SHRSP. Treatment of SHRSP with perindopril (2 mg/kg/day) for 6 weeks decreased systolic blood pressure and improved the ACh induced relaxation of aorta and carotid artery. The treatment inhibited the contraction by higher concentrations of ACh in the presence of L-nitroarginine. These results indicate that the impairment of endothelium-dependent relaxation in aorta and carotid artery of SHRSP may be caused by the reduced availability of nitric oxide. The perindopril-treatment may prevent these changes in SHRSP. PMID- 10830477 TI - Unique catalytic and molecular properties of hydrolases from Aspergillus used in Japanese bioindustries. AB - This review covers the unique catalytic and molecular properties of three proteolytic enzymes and a glycosidase from Aspergillus. An aspartic proteinase from A. saitoi, aspergillopepsin I (EC 3.4.23.18), favors hydrophobic amino acids at P1 and P'1 like gastric pepsin. However, aspergillopepsin I accommodates a Lys residue at P1, which leads to activation of trypsinogens like duodenum enteropeptidase. Substitution of Asp76 to Ser or Thr and deletion of Ser78, corresponding to the mammalian aspartic proteinases, cathepsin D and pepsin, caused drastic decreases in the activities towards substrates containing a basic amino acid residue at 1. In addition, the double mutant T77D/G78(S)G79 of porcine pepsin was able to activate bovine trypsinogen to trypsin by the selective cleavage of the K6-I7 bond of trypsinogen. Deuterolysin (EC 3.4.24.39) from A. oryzae, which contains 1g atom of zinc/mol of enzyme, is a single chain of 177 amino acid residues, includes three disulfide bonds, and has a molecular mass of 19,018 Da. It was concluded that His128, His132, and Asp164 provide the Zn2+ ligands of the enzyme according to a 65Zn binding assay. Deuterolysin is a member of a family of metalloendopeptidases with a new zinc-binding motif, aspzincin, defined by the "HEXXH + D" motif and an aspartic acid as the third zinc ligand. Acid carboxypeptidase (EC 3.4.16.1) from A. saitoi is a glycoprotein that contains both N- and O-linked sugar chains. Site-directed mutagenesis of the cpdS, cDNA encoding A. saitoi carboxypeptidase, was cloned and expressed. A. saitoi carboxypeptidase indicated that Ser153, Asp357, and His436 residues were essential for the enzymic catalysis. The N-glycanase released high-mannose type oligosaccharides that were separated on HPLC. Two, which had unique structures of Man10 GlcNAc2 and Man11GlcNAc2, were characterized. An acidic 1,2-alpha mannosidase (EC 3.2.1.113) was isolated from the culture of A. saitoi. A highly efficient overexpression system of 1,2-alpha-mannosidase fusion gene (f-msdS) in A. oryzae was made. A yeast mutant capable of producing Man5GlcNAc2 human compatible sugar chains on glycoproteins was constructed. An expression vector for 1,2-alpha-mannosidase with the "HDEL" endoplasmic reticulum retention/retrieval tag was designed and expressed in Saccharomyces cerevisiae. The first report of production of human-compatible high mannose-type (Man5GlcNAc2) sugar chains in S. cerevisiae was described. PMID- 10830478 TI - Autolysis of calpain large subunit inducing irreversible dissociation of stoichiometric heterodimer of calpain. AB - Calpain, a calcium dependent cysteine protease, consists of a catalytic large subunit and a regulatory small subunit. Two models have been proposed to explain calpain activation: an autolysis model and a dissociation model. In the autolysis model, the autolyzed form is the active species, which is sensitized to Ca2+. In the dissociation model, dissociated large subunit is the active species. We have reported that the Ca2+ concentration regulates reversible dissociation of subunits. We found further that in chicken micro/m-calpain autolysis of the large subunit induces irreversible dissociation from the small subunit as well as activation. So we could propose a new mechanism for activation of the calpain by combining our findings. Our model insists that autolyzed large subunit remains dissociated from the small subunit even after the removal of Ca2+ to keep it sensitized to Ca2+. This model could be expanded to other calpains and give a new perspective on calpain activation. PMID- 10830479 TI - Enzyme-linked immunosorbent assay for detection of chitooligosaccharides. AB - In order to detect chitooligosaccharides (COS), an enzyme-linked immunosorbent assay (ELISA) was developed. A chitooligosaccharide mixture (COSM) conjugated to bovine serum albumin was used to immunize rabbits to produce an anti-COS polyclonal antibody. By use of specific antibody and COSM-horseradish peroxidase conjugate, we established a competitive direct ELISA (cdELISA) the detection limit of which was about 0.1 microg/ml. In the cdELISA, the cross-reactivities of the specific antibody toward glucosamine, chitobiose, chitotriose, chitotetraose, chitopentaose, and chitohexaose were 0.27, 27, 75, 75, 144, and 100%, respectively, and those toward N-acetylchitobiose, N-acetylchitotriose, N acetylchitotetraose, N-acetylchitopentaose, and N-acetylchitohexaose were 1.58, 0.005, 1.08, 0.05, and 0.40%, respectively. PMID- 10830480 TI - Teasterone-3-O-beta-D-glucopyranoside, a new conjugated brassinosteroid metabolite from lily cell suspension cultures and its identification in lily anthers. AB - The new brassinosteroid conjugate, teasterone-3-O-betaD-glucopyranoside, was found as a metabolite of teasterone in lily cell suspension cultures. Its structure was determined by means of FAB-MS and 1H-NMR upon comparison with the authentic compound. Furthermore, its presence in lily anthers was confirmed by FAB-MS and LC-APCI-SIM data. This is the first natural brassinosteroid conjugate glucosylated at a hydroxyl group in ring A. PMID- 10830481 TI - Monitoring the irradiation-induced conformational changes of ovalbumin by using monoclonal antibodies and surface plasmon resonance. AB - Two types of conformationally specific anti-irradiated ovalbumin monoclonal antibodies were prepared in order to study and monitor irradiation-induced structural changes in the ovalbumin molecule. Surface plasmon resonance (SPR) detection was used to investigate the kinetic parameters of the reaction between antibodies and ovalbumin which had been administered with different doses of irradiation (0, 1.5, 2.0, 5.0, 10, 20, 50, and 100 kGy). The results demonstrate that the combination of monoclonal antibodies and the SPR method can be used to characterize the irradiation-induced conformational change with an unlabelled reagent. PMID- 10830482 TI - Molecular cloning and heterologous expression of pea seedling copper amine oxidase. AB - The cDNA coding for copper amine oxidase has been cloned from etiolated pea seedlings (Pisum sativum). The deduced amino acid sequence, consisting of 674 residues including the signal peptide, agreed well with those reported for the enzymes from a different cultivar of P. sativum and other plant sources, except for several evolutionary replacements located mostly on the molecular surface. A heterologous expression system for the cloned pea enzyme was constructed with the yeast Pichia pastoris, using the AOX1 promoter and the yeast alpha-factor secretion signal. Adding copper to the culture medium increased the secretion of an active, quinone-containing enzyme. Furthermore, the inactive enzyme produced in a copper-deficient medium was activated considerably by subsequent incubation with excess cupric ions. These results strongly suggest that the Tyr-derived redox cofactor, 2,4,5-trihydroxyphenylalanylquinone (topa quinone, TPQ), is produced in the plant enzyme by post-translational modification that proceeds through the copper-dependent, self-processing mechanism, as in the enzymes from bacteria and yeast. PMID- 10830483 TI - Comparison of chitinase isozymes from yam tuber--enzymatic factor controlling the lytic activity of chitinases. AB - To evaluate the anti-pathogen activity of chitinases, we developed a new method for measuring the lytic activity, and investigated the correlation of the lytic activity with the enzymatic properties by using four chitinase isozymes, Chitinases E, F, H1 and G, which had been purified from yam tubers by column chromatography. Chitinases E, F and H1 had high lytic activity against the plant pathogen, Fusarium oxysporum, but Chitinase G did not. Chitinase E, which is the family 19 chitinase, was similar to Chitinases F and G in its antigenecity, but not to Chitinase H1 or H2. Chitinases H1 and H2 were recognized by the anti Bombyx mori chitinase antibody, suggesting that Chitinases H1 and H2 are family 18 chitinases like B. mori chitinases. Chitinases E, F and H1 had two optimum pH ranges of 3-4 and 7.5-9 toward glycolchitin, but Chitinase G had only one optimum pH value of 5. Chitinases E, F and H1 had higher affinity to the polymer substrate, glycolchitin, than Chitinase G. These results suggest that the lytic activity of plant chitinases may be related to the chitin affinity and probably to the characteristic optimum pH value, or two values, but not related to its classification. The correlation of the lytic activity of a chitinase isozyme with its elicitor specificity is also discussed. PMID- 10830484 TI - Effects of branched cyclodextrins on the solubility and stability of terpenes. AB - In order to investigate the application potential of branched CDs, the solubilizing ability and the stabilizing ability of G2-betaCD and GUG-betaCD were investigated by using twelve terpenes (d-limonene, myrcene, terpinolene, geraniol, l-menthol, nerol, alpha-terpineol, citral, d-citronellal, l perillaldehyde, (R)-l-carvone, and menthone) as guest compounds. G2-betaCD and GUG-betaCD showed more solubilizing ability for these twelve terpenes than betaCD, and the ability of GUG-betaCD was almost the same as that of G2-betaCD. The stabilizing ability of terpene-GUG-betaCD complexes was different from that of G2-betaCD. GUG-betaCD was superior to G2-betaCD, especially in the solid state. This result may have been caused by the difference in structure of side chain, namely the hydroxymethyl group in G2-betaCD and the carboxyl group in GUG betaCD. PMID- 10830485 TI - Galactosylation of thiol group by beta-galactosidase. AB - beta-Galactosidase catalyzed beta-galactosylation not only of a hydroxyl group but also of a thiol group in the condensation reaction of D-galactose and 2 mercaptoethanol. The thio-galactosylation product was confirmed as 2-hydroxyethyl S-beta-D-galactoside on the bases of fast atom bombardment mass spectrometry, infrared spectroscopy, and nuclear magnetic resonance spectorometry. Aspergillus oryzae beta-galactosidase hydrolyzed p-nitrophenyl S-beta-D-galactoside most rapidly among several beta-galactosidases and produced the thio-galactosylation product most efficiently. The Penicillim multicolor enzyme was as effective as the A. oryzae enzyme. However the enzymes from Escherichia coli, Saccharomyces fragilis, Kluyveromyces lactis, and Bacillus circulans galactosylated hydroxyl groups predominantly to produce O-galactoside. The thio-galactoside was synthesized most effectively at a 2-mercaptoethanol concentration of about 1.25 M. Galactose concentration at 0.8-2.8 M did not affect the synthetic yield of the thiogalactoside so greatly. PMID- 10830486 TI - Syntheses of 4-methylumbelliferyl-beta-D-xylobioside and 5-bromo-3-indolyl-beta-D xylobioside for sensitive detection of xylanase activity on agar plates. AB - 4-Methylumbelliferyl-beta-D-xylobioside (MU-X2) and 5-bromo-3-indolyl-beta-D xylobioside (BI-X2) were synthesized as substrates for the detection of xylanase activity on agar plates. A family F/10 xylanase from Streptomyces olivaceoviridis E-86 (FXYN) was able to be more sensitively detected than RBB-xylan by using MU X2 as a substrate. A mutant xylanase E128H/FXYN having only 1/1000 of the activity of FXYN was also able to be detected on the MU-X2 plate but was not detected on the RBB-xylan plate. A family G/11 xylanase from Streptomyces lividans 66 (Xyn B) was not detected on the MU-X2 plate, but it was able to be detected on the RBB-xylan plate, suggesting that the MU-X2 substrate is specific to family F/10 xylanases. However, none of the xylanases were detected effectively by using BI-X2 as a substrate. PMID- 10830487 TI - Enzymatic production of trans-4-hydroxy-L-proline by regio- and stereospecific hydroxylation of L-proline. AB - A proline 4-hydroxylase gene, which was cloned from Dactylosporangium sp. RH1, was overexpressed in Escherichia coli W1485 on a plasmid under a tryptophan tandem promoter after the codon usage of the 5' end of the gene was optimized. The proline 4-hydroxylase activity was l600-fold higher than that in Dactylosporangium sp. RH1. trans-4-Hydroxy-L-proline(Hyp) was produced and accumulated to 41 g/L (87% yield from L-proline) in 100 h when the recombinant E. coli was cultivated in a medium containing L-proline and glucose. 2-Oxoglutarate, which is necessary for the hydroxylation of L-proline by proline 4-hydroxylase, was apparently supplied from glucose through the cellular metabolic pathway. The putA mutant of W1485, which is not able to degrade L-proline, has allowed the quantitative conversion of L-proline to Hyp. The formation of other isomers of hydroxyproline was not observed. Productivity of Hyp was almost the same in a larger-scale culture. The method of manufacturing Hyp from L-proline was established. PMID- 10830488 TI - Characterization of a phage resistance plasmid, pLKS, of silage-making Lactobacillus plantarum NGRI0101. AB - Phage contamination has resulted in abnormal fermentation in silage. We isolated a phage-resistant strain, Lactobacillus plantarum NGRI0101 from silage. The strain carried two plasmids, pLKL (6.8 kb) and pLKS (2.0 kb). By curing and retransformation of the plasmids, we clarified that pLKS has phage resistant activity, characterized as no adsorption inhibition. pLKS has 2,025 bp and three orfs, orfl23, orf132, and orf918. The predicted amino acid sequence of the orf918 product showed high similarity to those of Rep proteins of Pediococcus halophilus plasmid pUCL287 and Lactobacillus acidophilus plasmid pLA103. The replication origin (ori) was upstream from orf918. There was no gene similar to typical phage resistant genes encoded by known plasmids. The phage resistance of L. plantarum NGRI0101 may possibly be due to a plasmid-encoded abortive infection. PMID- 10830489 TI - Genetic characteristics of cellulose-forming acetic acid bacteria identified phenotypically as Gluconacetobacter xylinus. AB - Gluconacetobacter xylinus (=Acetobacter xylinum) shows variety in acid formation from sugars and sugar-alcohols. Toyosaki et al. proposed new subspecies of G. xylinus (=Acetobacter xylinum) subsp. sucrofermentans in point of acid formation from sucrose and a homology index of 58.2% with the type strain of G. xylinus subsp. xylinus in DNA-DNA hybridization experiments. We tried DNA-DNA hybridization to clarify relationship between acid formation from sugars and classification of G. xylinus. The G + C contents of G. xylinus showed 60.1-62.4 mol% with a range of 2.3 mol%. When type strains of G. xylinus subsp. xylinus, G. xylinus subsp. sucrofermentans, and IFO 3288 forming acid from sucrose, were used as probes, the DNAs from three strains showed 67-100%, 64-89%, and 60-100% similarity to those from sixteen strains including bacteria that form acid from sucrose or not. These results show that homology indexes do not reflect differences of acid formation from sucrose. As a results, the species G. xylinus was proved to be genetically homogeneous. PMID- 10830490 TI - Development of an improved assay for purine nucleoside kinase activity in cell extracts and detection of inosine kinase activity in Brevibacterium acetylicum ATCC 953, related species, and Corynebacterium flaccumfaciens ATCC 6887. AB - An improved assay was developed to detect direct purine nucleoside phosphorylating activity in cell-free extracts. Direct inosine phosphorylating activity was detected in 2 of 70 species tested. Both activities, which depended on magnesium ion and ATP, phosphorylated a hydroxyl group at the 5' position of inosine. The new assay was shown to be useful for screening of direct purine nucleoside phosphorylating activity and have the potential to detect inosine kinase in the presence of a background of nucleoside phosphorylase and purine phosphoribosyltransferase activities. Previously, the latter two activities made it difficult to correctly detect direct phosphorylation of inosine by inosine kinase. PMID- 10830491 TI - Some properties of a macromolecular conjugate of lysozyme prepared by modification with a monomethoxypolyethylene glycol derivative. AB - Hen egg-white lysozyme was modified with a succinyl ester derivative of monomethoxypolyethylene glycol (mPEG-COONSu), and some properties of the resulting conjugate (mPEG-lysozyme) were studied. The conjugate was prepared by modification of lysozyme with mPEG-COONSu and purified with use of columns of CM Toyopearl 650M and Sephadex G-75. Analytical data indicated that in the conjugate, 1.05 moles of mPEG with an average molecular weight of 5,000 were covalently attached to the lysozyme molecule. Tryptic peptide analysis of the conjugate showed that Lys 33 in lysozyme is the residue mainly modified with mPEG COONSu. Covalent attachment of the mPEG-derivative to amino groups greatly increased the thermostability of lysozyme without any conformational change of the protein molecule. mPEG-lysozyme retained full enzyme activity for glycol chitin, but lytic activity for Micrococcus luteus cells in neutral media was 75% of that of native lysozyme and its optimal pH was at pH 5.0. It was also found that the reactivity of lysozyme with anti-lysozyme antibody from BALB/c mice or human lymphocytes was decreased by modification with mPEG-COONSu. From these findings, it was suggested that mPEG-COONSu can be advantageously used for protein tailoring of lysozyme. PMID- 10830492 TI - Isolation of a 41-kDa protein with cell adhesion activity for animal tumor cells from the mushroom Hypsizigus marmoreus by affinity chromatography with type IV collagen immobilized on agarose. AB - A type IV collagen-binding protein of 41 kDa was isolated from the mushroom Hypsizigus marmoreus and the protein was designated as HM41. The Western blotting analysis with anti-HM41 antibodies demonstrated that HM41 was unrelated to HM23, which had been shown to have an affinity for type IV collagen. The microsequence analysis of the membrane-blotted peptides generated by fragmentation with cyanogen bromide showed no homologous proteins reported. HM41 had cell adhesion promoting activity for murine Lewis lung carcinoma LL2 cells and human fibrosarcoma HT1080 cells. These results indicate that HM41 is a hitherto undescribed fungus protein that can interact both with animal extracellular matrix protein type IV collagen and with animal tumor cells. PMID- 10830493 TI - Resolution and synthesis of optically active alcohols with immobilized water soluble proteins from green pea, soybean and buckwheat as new bio-catalysts. AB - Kinetic resolution of racemic alcohols, (+/-)-1-(4-substituted phenyl)ethanol and (+/-)-1-(2-naphthyl)ethanol, was done with immobilized green pea, soybean, or buckwheat proteins. The resolution was done stereoselectively by oxidizing only one enantiomer of a racemic alcohol to leave an optically active alcohol with a high purity. In addition, each protein could be reused consecutively at least three times without any decrease of yield or optical purity. PMID- 10830494 TI - An ELISA-based assay for detergent-solubilized cellular beta 1,4 galactosyltransferase activity. Use of a polyacrylamide derivative with GlcNAc beta side chains as a solid phase acceptor substrate. AB - In our previous paper [Oubihi et al. (1998) Anal. Biochem., 257, 169-175], we have shown that a polyacrylamide-derived synthetic glycopolymer with GlcNAcbeta side chains, termed PAP(GlcNAcbeta), is useful as a solid phase acceptor substrate for the ELISA-based analyses of soluble beta1,4(-)galactosyltransferase (GalT) activity in milk. This method is now used to assay detergent-solubilized cellular GalT. The glycopolymer coated on polystyrene plates was shown to be highly stable against the non-ionic and ionic detergents tested (0 approximately 5% solutions of Triton X-100 and SDS). Such stability made it possible to incubate the ELISA plate with detergent-solubilized GalT and to wash the ELISA plate with SDS solution after the GalT reaction, leading to high accuracy and sensitivity of this assay. The GalT activity was assayed using this method for 1% Triton X-100 extracts of various tissue samples of mice and several cultured cell lines. The results showed that the specific GalT activity of tissue extracts was low in brain and intestine, and high in ovary, muscle, and kidney. As for the cultured cell lines, COS7, COMMA-1D and C2C12 cells showed high specific activity, while CHO and MDCK cells showed low activity. The myoblast C2C12 had a slight increase in GalT activity during starvation-induced cell differentiation. On the other hand, GaIT-I transcript estimated by RT-PCR rather decreased during C2C12 cell differentiation, suggesting a differentiation-dependent switch in GalT isozymes. Taken all together, the ELISA-based assay using PAP(GlcNAcbeta) as a solid phase acceptor substrate was demonstrated to be a useful method for the assay of membrane-bound galactosyltransferases. PMID- 10830495 TI - Structured triacylglycerol containing medium-chain fatty acids in sn-1(3) facilitates the absorption of dietary long-chain fatty acids in rats. AB - A study was carried out to examine if the positional distribution of medium chain fatty acids (MCF) in triacylglycerol influences dietary fat absorption in rats. Two types of structure-specific fats, one predominantly composed of MCF in sn 1(3) and iinoleic acid in sn-2 [sn1(3)MCF-structured] and the others of MCF in sn 2 and linoleic acid in sn-1(3) [sn-2MCF-structured], were initially prepared, and the two structure-specific fats were interesterified and designated as sn-1(3)MCF interesterified and sn-2MCF-interesterified. Synthetic fat was mixed with an equal amount of cocoa butter (103 g/kg of diet) and was supplemented to the AIN93G-based diet. Rats were fed on the diets for 4 wk. Long-chain saturated fatty acids were the predominant fatty acids excreted into the feces, and the positional distribution of MCF resulted in an altered fat absorption rate (%) of 81.8, 82.5, 84.2 and 86.3 for the rats fed on the diets containing sn-2MCF structured, sn-1(3)MCF-interesterified, sn-2MCF-interesterified and sn-1(3)MCF structured fats, respectively. The proportion of MCF in the serum, liver and adipose tissue triacylglycerols was not affected by the MCF distribution of the dietary fats. These results indicate that the distribution of MCF in dietary triacylglycerol is a determinant of intestinal fat absorption. PMID- 10830496 TI - Deletion of the yhhP gene results in filamentous cell morphology in Escherichia coli. AB - The Escherichia coli yhhP gene was predicted to encode a small hypothetical protein of 81 amino acids, the cellular function of which is not known. To gain insight into the function of this uncharacterized YhhP protein, genetic and biochemical studies were done. We first tried to express and purify the YhhP protein to prepare an anti-YhhP antiserum. Western blotting showed that the hypothetical yhhP gene is indeed transcribed and translated as a minor cytoplasmic protein. YhhP-deficient (delta yhhP) cells formed colonies poorly on a rich medium (e.g., Luria-Bertani medium) containing a relatively low concentration of NaCl, while they can grow normally either in LB containing 3% NaCl or in a synthetic medium (e.g., M9-glucose). During exponential growth in rich medium, an early step of cell division was inhibited in delta yhhP cells, forming filaments. For the YhhP-deficient filamentous cells, the FtsZ-ring formation was analyzed with immunofluorescence microscopy. The FtsZ-ring formation did not occur normally in the delta yhhP filaments, although the filamentous cells contained the FtsZ protein at a certain level comparable to that in the wild-type cells. The ftsZ gene was found to function as a multicopy suppressor of the delta yhhP mutant. Another multicopy suppressor gene was identified as the dksA gene. Provided that either the ftsZ or dksA gene was introduced into the mutant cells with its multicopy state, the resulting transformants were capable of growing in rich medium, formed wild-type short rods. These results are discussed with regard to the presumed function of this ubiquitous protein. PMID- 10830497 TI - Synthesis and biological activities of 4-chloroindole-3-acetic acid and its esters. AB - 4-Chloroindole-3-acetic acid (4-Cl-IAA) and its esters were synthesized from 2 chloro-6-nitrotoluene as the starting material. The biological activities of 4-CI IAA and its esters were determined by four bioassays. Except for the tert-butyl ester, 4-Cl-IAA and its esters had stronger elongation activity toward Avena coleoptiles than had indole-3-acetic acid. The biological activities of the methyl, ethyl and allyl esters were as strong as the activity of the free acid. All the esters, except for the tert-butyl, inhibited Chinese cabbage hypocotyl growth more than the free acid did, and all the esters induced severe swelling and formation of numerous lateral roots in black gram seedlings even at a low concentration. Furthermore, adventitious root formation was strongly promoted in Serissa japonica cuttings by all the esters. The root formation-promoting activities of the ethyl and allyl esters were about three times the value for indole-3-butyric acid which is used to promote and accelerate root formation in plant cuttings. PMID- 10830499 TI - Mutational evidence supporting the involvement of tripartite residues His183, Asp185, and His243 in Streptomyces clavuligerus deacetoxycephalosporin C synthase for catalysis. AB - Deacetoxycephalosporin C synthase (DAOCS) is a non-heme iron-binding and alpha ketoglutarate dependent enzyme involved in catalyzing the biosynthesis of cephalosporins and cephamycins, antibiotics more potent than penicillins. In the crystal structure complex of Streptomyces clavuligerus DAOCS (scDAOCS), it was proposed that histidine-183, aspartate-185, and histidine-243 are putative iron binding ligands. However, coordinates proposed for crystal structures of proteins may not definitely comply with catalysis. Hence, site-directed mutagenesis was done to replace each of these amino acid residues with leucine. The constructed expression vectors bearing the mutations were found to express the respective scDAOCS mutant enzymes at high levels in Escherichia coli BL21(DE3). Through enzymatic assays, it was shown that while the wildtype enzyme could convert penicillin to a more active cephalosporin, the substitution of the three proposed iron-binding sites of scDAOCS completely abolished the same activity in the respective mutant enzymes. Thus, these results clearly indicate that histidine 183, aspartate-185, and histidine-243 of scDAOCS are essential for the ring expansion activity. PMID- 10830498 TI - Molecular cloning and characterization of a transcriptional activator gene, amyR, involved in the amylolytic gene expression in Aspergillus oryzae. AB - A gene, designated amyR, coding for a transcriptional activator involved in amylolytic gene expression has been cloned from Aspergillus oryzae by screening for a clone that enabled to reverse the reduced expression of the alpha-amylase gene (amyB) promoter. amyR encodes 604 amino acid residues of a putative DNA binding protein carrying a zinc binuclear cluster motif (Zn(II)2Cys6) belonging to the GAL4 family of transcription factors. The amyR gene disruptants showed a significant restricted growth on starch medium and produced little of the amylolytic enzymes including alpha-amylase and glucoamylase compared with a non disruptant, indicating that amyR is a transcriptional activator gene involved in starch/maltose-induced efficient expression of the amylolytic genes in A. oryzae. In addition, sequencing analysis found that amyR, agdA (encoding alpha glucosidase), and amyA (encoding alpha-amylase), are clustered on a 12-kb DNA fragment of the largest chromosome in A. oryzae, and that amyR is about 1.5 kb upstream of agdA and transcribed in the opposite direction. Furthermore, transcriptional analysis revealed that the amyR gene was expressed in the presence of glucose comparable to the level in the presence of maltose, while the amylolytic genes were transcribed at high levels only in the presence of maltose. PMID- 10830500 TI - Hybridization and breeding of the benomyl resistant mutant, Trichoderma harziantum antagonized to phytopathogenic fungi by protoplast fusion. AB - A diploid strain obtained from heterokaryons of Trichoderma harzianum by protoplast fusion grew on minimal medium containing 100ppm benomyl. This strain inhibited the growth of the phytopathogenic fungus Fusarium oxysporum f. sp. raphani on paired cultures and also protected against radish yellows and a drop in germination induced by F. oxysporum f. sp. raphani. PMID- 10830501 TI - Superoxide-scavenging and prolyl endopeptidase inhibitory activities of Bangladeshi indigenous medicinal plants. AB - The superoxide-scavenging and prolyl endopeptidase (PEP) inhibitory activities of 15 different kinds of Bangladeshi medicinal plants were evaluated. Methanol extraction was performed for the screening tests. Swertia chirata, Emblica officinalis, Zingiber officinale and Myristica malabarica were screened as superoxide-scavenging samples. Similarly, E. officinalis was identified as one of the strongest PEP inhibitory samples. The 50% (O2-)-scavenging and PEP-inhibitory concentrations from E. officinalis methanol extracts were found to be 13.17 and 26.10 microg/ml, respectively. PMID- 10830502 TI - Secreted phytase activities of yeasts. AB - The enzyme phytase dephosphorylates phytin (inositol hexaphosphate), a major phosphate reserve in plants. We found that a large number of yeast species secreted a phytase. Several species were identified as high phytase producers. The yeast enzymes had an optimal activity at pH 4-5 and generally a very high optimal temperature, ranging from 60 degrees C to 80 degrees C. PMID- 10830503 TI - Development of a simple and efficient method for transformation of buckwheat plants (Fagopyrum esculentum) using Agrobacterium tumefaciens. AB - Apical meristems of seedlings of buckwheat (Fagopyrum esculentum var. Shinano No. 1) were pricked with a needle and inoculated with Agrobacterium tumefaciens (LBA4404, pBI121). The inoculated seedlings were grown to maturation and allowed to pollinate randomly to set the seeds (T1 plants). The transformation efficiency of the T1 plants was estimated by germination in the presence of geneticin (20 microg/ml) and by detection of beta-glucuronidase (GUS) gene with PCR, indicating that 36% and 70% of the T1 plants were transformed, respectively. Four plants taking on a mutated morphology were selected from T1 plants which were transformed with the method using A. tumefaciens harboring a modified pBI121 for plasmid rescue. Southern blot analysis of plasmids rescued from the 4 T1 plants demonstrated that each plasmid contained a different flanking DNA of the buckwheat genome, an evidence that T-DNA was integrated in different sites of the genomic DNA among the 4 T1 plants. PMID- 10830504 TI - Dependence of rat spot14 promoter activity on NF-Y binding to the inverted CCAAT element at -100. AB - Electrophoretic mobility shift assay (EMSA) and in vitro transcription/translation show that NF-Y binds to the inverted CCAAT-element in the promoter of the rodent spot14 gene. The NF-Y-binding sequence has been shown to be responsible for basal activity in H4IIE. Given the similar role found for the inverted CCAAT-element in the promoter of the FAS gene, NF-Y may have an important function in the control of lipogenesis. PMID- 10830505 TI - Gene organization for nitric oxide reduction in Alcaligenes faecalis S-6. AB - norB and norC encoding the cytochrome b-containing subunit and the cytochrome c containing subunit, respectively, of the nitric oxide reductase (NOR) in Alcaligenes faecalis S-6 were cloned and sequenced. Both NorB and NorC showed more than 40% sequence identity to the corresponding subunits of cytochrome bc type NORs in other denitrifying bacteria. norCB was in a gene cluster containing seven other genes; these were named dnr, orf2, orf3, norE, norF, norQ, and norD on the basis of their similarity with NOR systems in other bacteria. Potential FNR-binding sites were present in front of norCB, norEF, and/or orf2/orf3, suggesting that most of these genes are regulated simultaneously by an FNR related protein. NorB and NorC proteins produced in the membrane fraction in Escherichia coli showed no enzyme activity, probably due to lack of NorQ and NorD, which appear to perform some essential function for activation of the NorB NorC complex in the recombinant E. coli. PMID- 10830506 TI - Distribution of sarcophytol A in soft coral of the Sarcophyton genus. AB - The distribution of sarcophytol-A in the Sarcophyton genus was investigated in seven samples belonging to S. glaucum (3 samples), S. infundibulifurme (2 samples), S. crassocaule (1 sample) and S. trocheliophorum (1 sample) that were collected on Ishigaki Island in Okinawa Prefecture. Sarcophytol-A was present in one sample each of S. glaucum and S. infundibulifurme. This study indicates that the composition of cembranoids in the Sarcophyton genus is not related with the respective species, but with the individual samples collected. PMID- 10830507 TI - Dehydrodimers of caffeic acid in the cell walls of suspension-cultured Mentha. AB - Dehydrodicaffeic acid derivatives were found in the cell walls of suspension cultured cells of Mentha. Using gas chromatography/mass spectrometry (GC-MS) in a single ion chromatography at m/z 790 and m/z 718, eleven peaks of trimethylsilylated dehydrodimers of caffeic acid were detected in the extracts from the cell walls of suspension-cultured cells of Mentha using sodium hydroxide. The result suggests that dehydrodicaffeates are formed in the cell walls from two molecules of caffeate, probably formed through C-C, and C-O-C coupling processes. PMID- 10830508 TI - Identification of L-inositol and scyllitol and their distribution in various organs in chrysanthemum. AB - Two unidentified soluble carbohydrates were isolated from chrysanthemum (Dendranthema x grandiflorum (Ramat.) Kitamura) leaves using HPLC. The compounds were identified as 1 L-chiro-inositol, called L-inositol (1) and scyllo-inositol, called scyllitol (2) from the results of 1H-NMR, 13C-NMR, and CI-MS spectra. L Inositol and scyllitol were distributed in four cultivars tested. L-Inositol concentration of petals gradually decreased during the flower bud development, but the L-inositol content increased by about 7 times. Scyllitol was detected only at an early stage of flower bud. PMID- 10830509 TI - New taxane diterpenoid from seed of the Chinese yew, Taxus yunnanensis. AB - A novel taxane diterpenoid with a rearranged 5/7/6-membered ring system was isolated from seeds of the Chinese yew, Taxus yunnanensis. Its structure was established as 9alpha,13alpha-diacetoxy-10beta-benzoxy-5alpha- cinnamoyl-11(1 5- >1)-abeotaxa-4(20),11-dien-15-ol on the basis of a spectroscopic analysis. Its relative stereochemistry is proposed from the results of NOESY experiments. PMID- 10830510 TI - A new chlorinated red naphthoquinone from roots of Sesamum indicum. AB - A new chlorinated red naphthoquinone pigment having antifungal activity, named chlorosesamone, was isolated from the roots of Sesamum indicum. Its structure was characterized as 2-chloro-5,8-dihydroxy-3-(3methyl-2-butenyl)- 1,4-na phthoquinone on the basis of spectral evidence. PMID- 10830512 TI - Stereoselective synthesis of the optically active samin type of lignan from L glutamic acid. AB - The optically active samin type of lignan, (1R,2S,5R, 6S)-6-(2-methoxy-4,5 methylenedioxyphenyl)-3,7-dioxabicyclo[3.3.0]octan-2-ol, was stereoselectively synthesized from L-glutamic acid via (2R,3R)-2-[(1S and R)1-[(tert - butyldimethylsilyl)oxyl-1-(2-methoxy-4,5methylenedioxyphenyl)methyl]-3-[(tert- butyldiphenylsilyl)oxylmethyl-1,4-butanediol. PMID- 10830511 TI - Steroid 5alpha-reductase inhibitory activity and hair regrowth effects of an extract from Boehmeria nipononivea. AB - The acetone extract of Boehmeria nipononivea showed both potent 5alpha-reductase inhibitory activity and hair regrowth promotion effects on mice. 5alpha-Reductase inhibitory activity-guided fractionation led to six active fatty acids: alpha linolenic, linoleic, palmitic, elaidic, oleic and stearic acids. The extract of B. nipononivea, and alphalinolenic, elaidic and stearic acids exhibited a hair regrowth effect. PMID- 10830513 TI - (10E,12Z,15Z)-9-hydroxy-10,12,15-octadecatrienoic acid methyl ester as an anti inflammatory compound from Ehretia dicksonii. AB - The methanol extract of Ehretia dicksonii provided (10E, 12Z, 15Z)-9-hydroxy 10,12,15-octadecatrienoic acid methyl ester (1) which was isolated as an anti inflammatory compound. Compound 1 suppressed 12-Otetradecanoyl-phorbol-13-acetate (TPA)-induced inflammation on mouse ears at a dose of 500 microg (the inhibitory effect (IE) was 43%). Linolenic acid methyl ester did not inhibit this inflammation at the same dose. However, the related compounds of 1, (9Z,11E) 13hydroxy-9,11-octadecadienoic acid (5) and (9Z,llE)13-oxo-9,11-octadecadienoic acid (6), showed potent activity (IE500 microg of 63% and 79%, respectively). Compounds 1, 4 ((9Z, 12Z, 14E)-16-hydroxy-9,12,14-octadecatrienoic acid), 5 and 6 also showed inhibitory activity toward soybean lipoxygenase at a concentration of 10 microg/ml. PMID- 10830514 TI - Purification and partial characterization of a basic xylanase produced by thermoalkaliphilic Bacillus sp. strain TAR-1. AB - A basic xylanase was purified from the culture supernatant of thermoalkaliphilic Bacillus sp. strain TAR-1. Its molecular mass and isoelectric point were 23 kDa and > pH 9.3, respectively. The enzyme showed a broad pH profile and was optimally active at 70 degrees C. Analyses of xylan-degradation products and N terminal amino acid sequence revealed that the enzyme would be a family 11/G endoxylanase. PMID- 10830515 TI - Effects of soybean saponin on protease hydrolyses of beta-lactoglobulin and alpha lactalbumin. AB - Effects of Soybean Saponin on Protease Hydrolyses of beta-Lactoglobulin and alpha Lactalbumin The effects of soybean saponin on tryptic and chymotryptic hydrolyses of whey proteins were evaluated. beta-lactoglobulin and alpha-lactalbumin became more sensitive to both trypsin and chymotrypsin by interacting with saponin in contrast to serum albumin. Soybean saponin was shown to have different effects on various proteins according to their nature. PMID- 10830516 TI - Two novel taxane diterpenoids from the needles of Japanese yew, Taxus cuspidata. AB - Two novel taxane diterpenoids were isolated from the needles of Japanese yew, Taxus cuspidata, and their structures were determined to be lbeta-hydroxy-7beta acetoxytaxinine (1) and lbeta,7beta-dihydroxytaxinine (2) on the basis of spectral analyses including 2D-NMR studies. PMID- 10830517 TI - Efficient selection for thermostable protease in Thermus thermophilus. AB - An efficient procedure was established to select for thermostable proteases in an extreme thermophile, Thermus thermophilus. A non-protease-secreting mutant derived from T. thermophilus TH125 was used as host and the expression plasmid for aqualysin I from T. aquaticus YT-1 was constructed as a source of thermostable protease. T. thermophilus cells harboring the recombinant plasmid produced active aqualysin I into the medium and were able to grow on a minimal medium containing milk casein as the sole source of carbon and nitrogen. PMID- 10830518 TI - The tyrosine kinase activity of the chicken insulin receptor is similar to that of the human insulin receptor. AB - The tyrosine kinase activity of a chimeric insulin receptor composed of the extracellular domain of the human insulin receptor (IR) and the intracellular domain of the chicken IR was compared with wild-type human IR. The degrees of autophosphorylation, phosphorylation of IRS-1, and in vitro phosphorylation of an exogenous substrate after stimulation by human insulin were similar to that seen with the human IR. We conclude that the insulin resistance of chickens is not attributable to a lower level of intrinsic tyrosine kinase activity of IR. PMID- 10830519 TI - (+)-4-epi-alpha-bisabolol as a major sesquiterepene constituent in the leaves of two Rosa rugosa hybrids, Martin Frobisher and Vanguard. AB - During an investigation of the sesquiterpene phases and contents in leaves of several Rosa rugosa hybrids (hybrid rugosas), Martin Frobisher and Vanguard were found to accumulate a large amount of (+)-4-epi-alphabisabolol (1) as a single constituent. Although glandular trichomes of Martin Frobisher on the leaves are dense, this R. rugosa hybrid produces none of the carota-1,4dienaldehyde (2) or bisaborosaol A (3) that are both found as representative sesquiterpenes of the carotane and bisabolane classes, respectively, in a glandular trichome exudate of wild-type R. rugosa. Compound 1 was also apparent as a nearly single constituent detectable by GC in the leaf constituents of Vanguard possesses sparse glandular trichomes on the leaf. Martin Frobisher and Vanguard had likely lost their capability to form carotane-type sesquiterpenes and had also lost their activity to oxygenate the C-7 allyl methyl carbon of compound 1 to convert 3. The presence of (+)-4-epi-alphabisabolol-accumulating R. rugosa hybrids is significant when considering the sesquiterpene biogenesis of Rosa rugosa. PMID- 10830520 TI - Galacturonic-acid-induced increase of superoxide production in red tide phytoplankton Chattonella marina and Heterosigma akashiwo. AB - Red tide phytoplankton, Chattonella marina and Heterosigma akashiwo, are known to generate superoxide anion (O2-). We found that galacturonic acid (GaLUA) stimulated C. marina and H. akashiwo to generate increased amounts of O2-. Since such effect was not observed in any other monosaccharides tested, our results suggest that the binding of GalUA to specific sites on the flagellate cell surface may induce the increase of 02- production. PMID- 10830521 TI - Structural analysis of the ntrBC genes of deep-sea piezophilic Shewanella violacea. AB - The ntrBC genes coding for the bacterial signal-transducing protein NtrB and the bacterial enhancer-binding protein NtrC of deep-sea piezophilic Shewanella violacea were cloned and their nucleotide sequences were analyzed. The conserved regions of NtrB and those of NtrC are well conserved in the case of the ntrBC products of S. violacea. PMID- 10830522 TI - Identification and characterization of a potent antibacterial agent, NH125 against drug-resistant bacteria. AB - New imidazole compounds were synthesized to develop a novel and effective antibacterial agent (1-benzyl-3-cetyl-2-methylimidazolium iodide, NH125). In vitro experiments demonstrated that NH125 effectively inhibited a number of different histidine protein kinases. Furthermore, oxacillin-resistant Staphylococcus aureus (ORSA), vancomycin-resistant Enterococcus faecalis (VRE), penicillin-resistant Streptococcus pneumoniae (PRS), and other Gram-positive and Gram-negative bacteria were found to be very sensitive to NH125. PMID- 10830523 TI - Assessment of left ventricular function by gated myocardial perfusion and gated blood-pool SPECT: can we use the same reference database? AB - The purpose of this study was to compare left ventricular (LV) volume and ejection fraction (LVEF) measurements obtained with electrocardiographic gated single-photon emission computed tomographic (SPECT) myocardial perfusion imaging (GS-MPI) with those obtained with gated SPECT cardiac blood-pool imaging (GS pool). Fifteen patients underwent GS-MPI with technetium-99m-tetrofosmin and GS pool with technetium-99m-erythrocyte, within a mean interval of 8 +/- 3 days. Eight patients had suspected dilated cardiomyopathy and seven patients had angiographically significant coronary artery disease. End-diastolic volume (EDV), end-systolic volume (ESV) and LVEF measurements were estimated from GS-MPI images by means of Cedars-Sinai automatic quantitative program and from GS-pool images by the threshold technique. Mean differences between GS-MPI and GS-pool in EDV, ESV and LVEF measurements were -2.8 +/- 10.5 ml [95% confidence interval (CI): 8.6 +/- 3.0 ml], 2.6 +/- 7.3 ml (CI: -1.4 +/- 6.6 ml) and -2.3 +/- 5.1% (CI: -5.1 +/- 0.6%), respectively. No significant difference in the mean differences from 0 was found for EDV, ESV or LVEF measurements. Bland-Altman plots revealed no trend over the measured LV volumes and LVEF. For all parameters, regression lines approximated lines of identity. The excellent agreement between GS-MPI and GS pool measurements suggests that, for estimation of LV volumes and LVEF, these two techniques may be used interchangeably and measurements by one method can serve as a reference for the other. PMID- 10830524 TI - Further characterization of a CNS adenosine A2a receptor ligand [11C]KF18446 with in vitro autoradiography and in vivo tissue uptake. AB - PET assessment of the adenosine A2a receptors localized in the striatum offers us a potential new diagnostic tool for neurological disorders. In the present study, we carried out in vitro receptor autoradiography of a newly developed PET ligand [11C]KF18446 ([7-methyl-11C]-(E)-8-(3,4,5-trimethoxystyryl)-1,3,7 trimethylxanthin e) with rat brain sections. [11C]KF18446 showed a high striatum/cortex binding ratio (5.0) and low nonspecific binding (<10%), suggesting that [11C]KF18446 has characteristics comparable or slightly superior to [3H]CGS 21680 or [3H]SCH 58261, which are currently available representative A2a receptor ligands. Scatchard analysis indicated a Kd of 9.8 nM and a Bmax of 170 fmol/mm3 tissue in the striatum and a Kd of 16.4 nM and a Bmax of 33 fmol/mm3 tissue in the cortex. Seven xanthine-type and four nonxanthine-type adenosine receptor ligands with an affinity for the adenosine A2a receptors significantly reduced the in vitro binding of [11C]KF18446 to the brain section. The blocking effects were much stronger in the striatum than in the cortex, but did not necessarily parallel their affinity. On the other hand, four xanthine-type ligands and one nonxanthine-type ligand (SCH 58261) of the 11 ligands studied reduced the in vivo uptake of [11C]KF18446 in mice, but other ligands, including A1-selective and nonselective ligands and three nonxanthine-type A2a-selective antagonists did not. We conclude that [11C]KF18446 is a promising adenosine A2a receptor ligand for PET study. PMID- 10830525 TI - Thallium-201 SPECT with triple-headed gamma camera for differential diagnosis of small pulmonary nodular lesion 20 mm in diameter or smaller. AB - AIM: Although thallium-201 (201Tl) has been used for the diagnosis of lung cancer, its detectability of small pulmonary nodules is not known. The aim of this study was to evaluate the ability of 201Tl SPECT for the differential diagnosis for the pulmonary nodules 20 mm in diameter or smaller. METHODS: 201Tl SPECT was performed in 31 patients suspected of having primary lung cancer. The final diagnosis was established by histology, and tumor size was 10 to 20 mm in diameter. Twenty of 31 patients had malignant tumors, including squamous cell lung cancer (n = 5), adenocarcinoma (n = 14) and small cell lung cancer (n = 1), but in none of them was there mediastinal lymphnode involvement. RESULTS: Ten of 20 malignant tumors and 1 of 11 benign lesions demonstrated significant 201Tl uptake, so that the positive predictive value, negative predictive value, sensitivity and specificity for the diagnosis of lung cancer were 90.9% (10/11), 50.0% (10/20), 50.0% (10/20) and 90.9% (10/11), respectively. CONCLUSION: These data suggest that sensitivity for detecting lung cancer 20 mm or less in diameter may be insufficient, but even in patients with small pulmonary nodules, a positive 201Tl result is highly predictive of lung cancer. PMID- 10830526 TI - Sex-related differences in the muscarinic acetylcholinergic receptor in the healthy human brain--a positron emission tomography study. AB - We evaluated the sex-related differences in the decline of the cerebral muscarinic acetylcholinergic receptor (mACh-R) due to aging by using 11C-N-methyl 4-piperidyl benzilate (11C-NMPB) and positron emission tomography (PET). The subjects consisted of 37 (20 males and 17 females) healthy volunteers. The 11C NMPB uptake was evaluated by the ratio method (regional 11C-NMPB uptake/Cerebellar 11C-NMPB uptake; rNMPB ratio). The correlation between sex, aging, and the rNMPB ratio in normal aging was evaluated by a multiple regression analysis. The rNMPB ratio was higher in females than in males throughout the entire cerebral region (p < 0.01-p < 0.0001) and the rNMPB ratio might thus possibly decline with age more rapidly in females. Our study therefore revealed the existence of sex-related differences in the cerebral mACh-R. PMID- 10830527 TI - The usefulness of 99mTc-HMPAO-labeled leukocyte scintigraphy in the diagnosis of skeletal metastases of cancers. AB - The usefulness of bone marrow scintigraphy with 99mTc-HMPAO-labeled leukocytes (leukocyte bone marrow scintigraphy) in the diagnosis of skeletal metastases of cancers was investigated in 70 lesions in 27 patients with various types of cancer. The final diagnosis of skeletal metastases was based on one or more criteria consisting of histological confirmation, typical findings of metastases by bone radiograph, CT and MRI, or progressive swellings of the lesions with severe pain due to nerve compression. Of the 70 lesions, 55 were finally diagnosed as metastases, and 15 as benign lesions. Leukocyte bone marrow scintigraphy showed photopenic defects in 52 of the 55 metastatic lesions (sensitivity 95%), and the remaining 3 negative lesions were found positive for metastases by MRI. In contrast, MRI could evaluate only 39 of the 55 lesions because 16 lesions in the ribs, scapula and sternum were not visualized. Of these 39 lesions, MRI showed positive findings for metastases in 33 (sensitivity 85%), and negative findings in 6 with photopenic defects found by leukocyte bone marrow scintigraphy. Of the 15 benign lesions, 3 were false positive for metastases on leukocyte bone marrow scintigraphy (specificity 80%). We conclude that 99mTc HMPAO-labeled leukocyte bone marrow scintigraphy may be useful in the diagnosis of skeletal metastases of cancers, particularly when MRI fails to evaluate the lesions. PMID- 10830528 TI - Brain metastasis from differentiated thyroid cancer in patients treated with radioiodine for bone and lung lesions. AB - Brain metastasis of differentiated thyroid cancer (DTC) often is detected during treatment of other remote lesions. We examined the prevalence, risk factors and treatment outcome of this disease encountered during nuclear medicine practice. Of the 167 patients with metastasis to lung or bone treated 1-14 times with radioactive iodine (RAI), 9 (5.4%) also had lesions in the brain. Five were males and 4 females, aged 49-84, out of the original population of 49 males and 118 females aged 10-84 (mean 54.7) years. Three of them underwent removal of their brain tumors, 5 received conventional external beam irradiation, and 2 had stereotactic radiosurgery with supervoltage X-ray. None of the brain lesions showed significant uptake of RAI despite demonstrable accumulation in most extracerebral lesions. Seven patients died 4-23 (mean 9.4) months after the discovery of cerebral metastasis, brain damage being the primary or at least a contributing cause. The 8th and 9th patients remained relatively well for more than 42 and 3 months, respectively, without any evidence of intracranial recurrence. Our results confirmed that the brain is a major site of secondary metastasis from DTC. No statistically significant demographic risk factor was detected. Any suspicious neurological symptoms in the course of RAI treatment warrant cerebral computed tomography. As for therapy, from our initial experience, radiosurgery seemed promising as an effective and less invasive alternative to surgical removal. PMID- 10830529 TI - A comparative study of simple methods to quantify cerebral blood flow with acetazolamide challenge by using iodine-123-IMP SPECT with one-point arterial sampling. AB - The aim of this study was to compare the accuracy of simplified methods for quantifying rCBF with acetazolamide challenge by using 123I-N-isopropyl-p iodoamphetamine (IMP) and SPECT with one-point arterial sampling. After acetazolamide administration we quantified rCBF in 12 subjects by the following three methods: (a) the modified microsphere method, (b) the IMP-autoradiographic (ARG) method based on a two-compartment one-parameter model, and (c) the simplified method based on a two-compartment two-parameter model (functional IMP method). The accuracy of these methods was validated by comparing rCBF values with those obtained by the standard method: the super-early microsphere method with continuous withdrawal of arterial blood. On analyzing rCBF in each flow range (0-0.25, 0.25-0.5, 0.5-0.75 and more than 0.75 ml/g/min), rCBF values obtained by both methods (a) and (c) showed significant correlations (p < 0.01) with those obtained by the standard method in every range, but rCBF values obtained by method (b) did not significantly correlated in the high flow range (0.5-0.75 and more than 0.75 ml/g/min). Method (c) was found to be the most accurate, even though it needs two serial SPECT scans. When requiring one SPECT scan, method (a) was considered to be superior to method (b) because of its accuracy, especially in high flow regions loaded with acetazolamide. PMID- 10830530 TI - Clinical usefulness of positron emission tomography with fluorine-18 fluorodeoxyglucose in the diagnosis of liver tumors. AB - We studied various liver tumors by positron emission tomography with fluorine-18 fluorodeoxyglucose (FDG-PET) to examine the diagnostic usefulness of this technique. We also examined the relation between findings on FDG-PET and the characteristics of hepatocellular carcinoma. FDG-PET was performed in 78 patients with liver tumors, including 53 with primary liver cancer [48 hepatocellular carcinomas (HCC) and 5 cholangiocellular carcinomas (CCC)], 20 with metastatic liver cancer, 2 with liver hemangioma, and 3 with focal nodular hyperplasia. For quantitative evaluation, a region of interest (ROI) was placed over the entire tumor region, at the level of the maximum diameter of the tumor. A background ROI was then placed over the non-tumor region of the liver. The average activity within each ROI was subsequently corrected for radioactive decay, and the standardized uptake value (SUV) was calculated by dividing the tissue activity by the injected dose of radioactivity per unit body weight. SUV ratio was expressed as the tumor-to-non-tumor ratio of the SUV. The median SUV was significantly lower in HCC than in metastatic live cancer or CCC, and the median SUV ratio was significantly lower in HCC than in metastatic liver cancer or CCC. The median SUV was not higher in multiple HCC than in single HCC, but the median SUV ratio was significantly higher in multiple HCC than in single HCC. The median SUV and the median SUV ratio were significantly higher in the presence of portal vein thrombosis than in the absence of such thrombosis. The Cancer of the Liver Italian Program score and the alpha-fetoprotein value correlated significantly with both the SUV and SUV ratio. These results suggest that FDG-PET is clinically useful not only for the differential diagnosis of liver tumors but also for evaluation of the clinical characteristics of HCC. PMID- 10830531 TI - Biodistribution and breast tumor uptake of 16alpha-[18F]-fluoro-17beta-estradiol in rat. AB - To evaluate the usefulness of 16alpha-[18F]-fluoro-17beta-estradiol (FES) for the assessment of estrogen receptor (ER), we examined the tissue distribution and kinetics of FES in immature female Sprague-Dawley rats and then examined FES uptake in rat breast tumors induced by 7,12-dimethylbenz(a) anthracene (DMBA). The FES uptake by the uterus, an ER-rich tissue, was highly selective and it was 3.34 +/- 0.79%ID/g at 60 minutes and 1.57 +/- 0.57%ID/g at 120 minutes after injection. The FES uptakes in ER-negative tissues were 0.12 +/- 0.05%ID/g or less and 0.05 +/- 0.03%ID/g or less, respectively. Coadministration of unlabeled beta estradiol showed marked depression of uterine FES uptake. The FES uptake by rat breast tumors was 0.14 +/- 0.06%ID/g at 60 min and 0.12 +/- 0.09%ID/g at 120 min. The FES uptake by rat breast tumors correlated with the ER concentration (r = 0.45, p < 0.05). In conclusion, these results suggest that the FES uptake by tissue is mainly ER mediated and FES is thus useful for detecting ER positive breast tumors. PMID- 10830532 TI - Protein-loss into retroperitoneal lymphangioma: demonstration by lymphoscintigraphy and blood-pool scintigraphy with Tc-99m-human serum albumin. AB - A rare, benign congenital lymphangioma has been reported to occur frequently in the neck and axilla, but rarely in the retroperitoneal space. We report a case of a retroperitoneal lymphangioma associated with hypoproteinemia caused by protein loss into the tumor. In this case, lymphoscintigraphy with subcutaneously injected Tc-99m-human serum albumin (HSA) disclosed the communication between the tumor and the lymphatic system, and sequential abdominal scintigraphy with intravenously injected Tc-99m-HSA revealed the protein loss into the tumor. Abdominal scintigraphy with Tc-99m-HSA injected intravenously or subcutaneously is occasionally useful for determining the etiology of hypoproteinemia. PMID- 10830533 TI - Discordant uptake of Tl-201 and Tc-99m MIBI in a patient with follicular adenoma. AB - Two phase Tl-201 and Tc-99m MIBI thyroid scintigraphies were carried out in a 30 year-old woman who had a solitary cold thyroid nodule seen on a pertechnetate scan. Although an early Tl-201 thyroid image showed intense uptake in the nodule, Tc-99m MIBI demonstrated a hypoactive lesion on the early image. Delayed thyroid scans showed faster washout from the nodule compared to normal thyroid tissue for both Tl-201 and Tc-99m MIBI. Later on, the patient was operated on and the nodule was found to be a follicular adenoma by histopathological diagnosis. PMID- 10830534 TI - Increased uptake of 99mTc-HL91 in tumor cells exposed to X-ray radiation. AB - 99mTc-HL91, a hypoxic marker, may be a predictor of tumor response to radiotherapy and an indicator of tumor oxygenation in the course of treatment. In this study, serial changes in 99mTc-HL91 uptake were observed in the normoxic condition in a human bladder cancer cell line exposed to a single dose or a fractionated dose of 10 Gy with an x-ray beam. The uptake per cell increased during cell growth retardation induced by the irradiation. This finding indicates that 99mTc-HL91 uptake is affected by injury to cells due to radiation; it may therefore be difficult to correctly assess the tissue oxygenation status during radiotherapy with 99mTc-HL91. PMID- 10830535 TI - The prognostic significance of angiogenesis in canine mammary tumors. PMID- 10830536 TI - Diagnosis of emerging ehrlichial diseases of dogs, horses, and humans. PMID- 10830538 TI - Chronic idiopathic inflammatory bowel diseases of the horse. AB - A review of reported cases of inflammatory bowel diseases (IBDs) of horses for which no etiology was identified included cases of granulomatous enteritis (GE), multisystemic eosinophilic epitheliotropic disease (MEED), lymphocytic plasmacytic enterocolitis (LPE), and idiopathic eosinophilic enterocolitis (EC). The terms EC and MEED were both used to describe a disease in horses characterized by infiltration of intestine and extraintestinal tissues with eosinophils. We use EC to describe IBD characterized by only intestinal infiltration by eosinophils. Horses with GE, MEED, or LPE are usually examined because of weight loss and depression, but horses with EC are usually examined because of signs of abdominal pain. Typically, horses with IBD have low concentrations of serumal proteins, especially albumin, and fail to adequately absorb glucose or xylose. Antemortem diagnosis of IBD can only be made by histologic examination of affected intestine. In some cases, antemortem diagnosis is made from histologic examination of rectal mucosa obtained by biopsy. Suspected causes of IBD in the horse include abnormal immune response to bacterial, viral, parasitic, or dietary antigens. Most horses with IBD do not survive, but horses with EC are more likely than those with LPE, MEED, or GE to respond to treatment. Successful treatments of horses with IBD include resection of grossly affected intestine and administration of corticosteroids. PMID- 10830537 TI - Ehrlichia equi infection of horses from Minnesota and Wisconsin: detection of seroconversion and acute disease investigation. AB - Equine granulocytic ehrlichiosis (EGE) is caused by infection with Ehrlichia equi. EGE has been reported primarily in northern California, where E equi is transmitted by the tick Ixodes pacificus. Reports of EGE and the emergence of human granulocytic ehrlichia in Minnesota prompted a seroprevalence study of E equi in horses of Minnesota and Wisconsin. Tick (Ixodes scapularis) endemic areas of Minnesota and Wisconsin were compared to nonendemic regions of Minnesota. Indirect fluorescent antibody was used to detect the presence of serum antibodies to E equi. Serum samples from healthy horses, 375 samples from I scapularis endemic counties, and 366 samples from nonendemic counties were screened at a 1:40 dilution. Results demonstrated a seroprevalence of 17.6% in endemic areas versus 3.8% in nonendemic areas. Ehrlichial DNA from 2 samples was successfully amplified by polymerase chain reaction and 919 base pairs were sequenced. The DNA sequence of 1 Minnesota/Wisconsin strain differed from the GenBank strain (M73223) of E equi at positions 84 and 886 and from the MRK strain of E equi at position 84, and was identical to the human granulocytic ehrlichiosis (HGE) agent. The 2nd Minnesota/Wisconsin strain was identical to the 1st with the exception of a substitution of "A" at position 453 that is not present in E phagocytophila, E equi, or HGE agent strain sequences. Based on the results of this study, we concluded that E equi is present and causes infection in horses in Minnesota and Wisconsin. The occurrence of infection is higher in tick endemic regions. PMID- 10830539 TI - Effect of spaying and timing of spaying on survival of dogs with mammary carcinoma. AB - The risk of developing mammary gland tumors in dogs is significantly decreased by ovariohysterectomy at an early age. However, previous studies have not found a benefit to ovariohysterectomy concurrent with tumor removal in dogs with established mammary gland tumors, suggesting that the progression of these tumors is independent of continued estrogen stimulation. The purpose of this study was to evaluate the effect of spaying and of the timing of spaying on survival in dogs with mammary gland carcinoma. Signalment, spay status and spay age, tumor characteristics, treatment. survival, and cause of death of 137 dogs with mammary gland carcinoma were analyzed. The dogs were classified into 3 groups according to spay status and spay time: intact dogs, dogs spayed less than 2 years before tumor surgery (SPAY 1), and dogs spayed more than 2 years before their tumor surgery (SPAY 2). Dogs in the SPAY 1 group lived significantly longer than dogs in SPAY 2 and intact dogs (median survival of 755 days, versus 301 and 286 days, respectively, P = .02 and .03). After adjusting for differences between the spay groups with regard to age, histologic differentiation, and vascular invasion, SPAY 1 dogs survived 45% longer compared to dogs that were either intact or in the SPAY 2 group (RR = .55; 95% CI .32-.93; P = .03). This study reveals ovariohysterectomy to be an effective adjunct to tumor removal in dogs with mammary gland carcinoma and that the timing of ovariohysterectomy is important in influencing survival. PMID- 10830540 TI - Evaluation of ifosfamide for treatment of various canine neoplasms. AB - lfosfamide (3-[2-chloroethyl]-2[(2 chloroethyl)amino]tetrahydro-2H-1,3,2 oxazaphosphorine 2-oxide) is an alkylating agent with a broad spectrum of antitumor activity. The efficacy and toxicity of ifosfamide were evaluated in 72 dogs with spontaneously occurring tumors. Forty dogs (56%) had lymphoma, 31 (43%) had sarcomas, and 1 had a metastatic carcinoma. Five dogs received ifosfamide at dosages <350 mg/m2 IV. Neither toxicity nor response were observed, and the remaining dogs received ifosfamide at 350 mg/m2 (n = 18) and 375 mg/m2 body surface area IV (n = 49). Saline diuresis and the thiol compound mesna were used to prevent urothelial toxicity. Fifty-two dogs had measurable tumors and could be evaluated for response. Complete responses were seen in 1 dog with metastatic leiomyosarcoma of the urinary bladder and in 1 dog with metastatic cutaneous hemangiosarcoma. One dog with lymphoma had a partial response for 112 days. Six dogs with splenic hemangiosarcoma received ifosfamide postsplenectomy and their median survival time was 147 days. The acute dose limiting toxicity was neutropenia 7 days after administration of ifosfamide. The median and mean neutrophil counts 7 days after ifosfamide at 350 mg/m2 were 2,035 cells/microL and 4,773 cells/microL, respectively (n = 12). The median and mean neutrophil counts 7 days after ifosfamide at 375 mg/m2 were 2,500 cells/microL and 3,594 cells/microL, respectively (n = 37). No dog developed clinical or microscopic evidence of hemorrhagic cystitis. Ifosfamide appears safe to use in tumor-bearing dogs, and the evaluation of combination chemotherapy protocols that include ifosfamide should be considered. PMID- 10830541 TI - Thyroid function and serum hepatic enzyme activity in dogs after phenobarbital administration. AB - Phenobarbital is the drug of choice for control of canine epilepsy. Phenobarbital induces hepatic enzyme activity, can be hepatotoxic, and decreases serum thyroxine (T4) concentrations in some dogs. The duration of liver enzyme induction and T4 concentration decreases after discontinuation of phenobarbital is unknown. The purpose of this study was to characterize the changes in serum total T4 (TT4), free T4 (FT4), thyroid-stimulating hormone (TSH), cholesterol and albumin concentrations, and activities in serum of alanine aminotransferase (ALT), alkaline phosphatase (ALP), and gamma-glutamyl transferase (GGT) after discontinuation of long-term phenobarbital administration in normal dogs. Twelve normal dogs were administered phenobarbital at a dosage of approximately 4.4-6.6 mg/kg PO q12h for 27 weeks. Blood was collected for analysis before and after 27 weeks of phenobarbital administration and then weekly for 10 weeks after discontinuation of the drug. The dogs were clinically normal throughout the study period. Serum ALT and ALP activity and TSH and cholesterol concentrations were significantly higher than baseline at week 27. Serum T4 and FT4 were significantly lower. Serum albumin and GGT were not changed from baseline at week 27. Changes in estimate of thyroid function (TT4, FT4, TSH) persisted for 1-4 weeks after discontinuation of phenobarbital, whereas changes in hepatic enzyme activity (ALT, ALP) and cholesterol concentration resolved in 3-5 weeks. To avoid false positive results, it is recommended that thyroid testing be performed at least 4 weeks after discontinuation of phenobarbital administration. Elevated serum activity of hepatic enzymes 6-8 weeks after discontinuation of phenobarbital may indicate hepatic disease. PMID- 10830542 TI - Eosinophilic bronchopneumopathy in dogs. AB - Eosinophilic bronchopneumopathy was diagnosed in 23 young dogs. Clinical signs included cough, gagging, and retching in all dogs, dyspnea in 21 dogs (91%), and nasal discharge in 12 dogs (52%). The most common radiographic findings were a moderate to severe bronchointerstitial pattern (68%, 13 of 19 dogs). Bronchoscopic findings included the presence of abundant yellow-green mucus or mucopurulent material (70%, 16 of 23 dogs) and severe mucosal thickening with an irregular or polypoid appearance (52%, 12 of 23 dogs), with partial airway closure during expiration in 3 dogs (13%). Peripheral blood eosinophilia was noted in 14 of 23 dogs (61%). Inflammatory cells in brush or bronchoalveolar lavage fluid cytologic preparations comprised more than 50% eosinophils in 14 of 23 dogs (61%), and 20-50% eosinophils in 6 dogs (26%). Eosinophilic infiltration of the bronchial mucosa was observed in biopsies from 19 dogs and was graded as mild (37%, 7 dogs), moderate (32%, 6 dogs), or severe (32%, 6 dogs). The mean serum immunoglobulin A concentration was almost double that of a population of 20 healthy dogs of various breeds. Oral glucocorticoids were administered on alternate days with progressive tapering of the dose; the dosage at maintenance varied between 0.1 and 1.0 mg/kg every other day. No relationship was found between the duration of clinical signs and the maintenance dosage or the cytologic and histopathologic grades. PMID- 10830543 TI - Proteins invoked by vitamin K absence and clotting times in clinically ill cats. AB - The clinical utility of the Thrombotest, a method for determining the prothrombin time that is uniquely sensitive to the presence of proteins invoked by vitamin K absence (PIVKA), was prospectively evaluated and compared to routine coagulation tests in cats with clinically suspected bleeding tendencies. Abnormal PIVKA clotting values were determined by comparison to results of a concurrently evaluated pooled feline plasma sample and by use of an absolute cutoff value of 25.2 seconds. To be recognized as abnormal, PIVKA clotting values had to be >20% of the pooled feline plasma PIVKA clotting time (the "20% rule") or > or =25.2 seconds (mean + 2 standard deviations of 150 different pooled feline plasma samples). Among the disorders in the population examined were 74 cats with liver disease and 19 cats with severe inflammatory bowel disease. Overall, a prolonged PIVKA clotting time based on the 25.2-second cutoff was found in 39.3% of cats, and based on the 20% rule in 40.7% of cats. An abnormal prothrombin time (PT) developed in 5.8% of cats, an abnormal APTT in 14% of cats, subnormal fibrinogen in 8.8% of cats, and thrombocytopenia in 3.3% of cats. Bleeding tendencies were confirmed in 22 cats, of which abnormal PIVKA clotting times were recognized in 95.5%, abnormal PT in 21%, abnormal activated partial thromboplastin time in 25%, hypofibrinogenemia in 16.7%, and thrombocytopenia in 4.5%. Response to treatment with vitamin K was demonstrated in 21 of 24 cats with an abnormal PIVKA clotting time. In these cats, an abnormal PIVKA clotting time normalized within 3 to 5 days of parenteral vitamin K administration. Cats responding to vitamin K administration had hepatic lipidosis (n = 7), severe inflammatory bowel disease (n = 4), severe inflammatory bowel disease associated with cholangiohepatitis (n = 5), and miscellaneous disorders (n = 5). Using either endpoint, the PIVKA clotting time is more sensitive for the detection of cats with coagulopathies than routinely used coagulation assessments in our hospital. Our findings confirm that cats with hepatic lipidosis, severe cholangiohepatitis, and severe inflammatory bowel disease develop coagulopathies responsive to vitamin K administration. PMID- 10830544 TI - Intestinal crypt lesions associated with protein-losing enteropathy in the dog. AB - Six dogs were diagnosed with protein losing enteropathy (PLE). There was no evidence of inappropriate inflammatory infiltrates or lymphangiectasia in multiple mucosal biopsies of the small intestine of 4 of the dogs. The 5th and 6th dogs had obvious lymphangiectasia and a moderate infiltrate of inflammatory cells in the intestinal mucosa. All 6 dogs had a large number of dilated intestinal crypts that were filled with mucus, sloughed epithelial cells, and/or inflammatory cells. Whether PLE occurs in these dogs because of protein lost from the dilated crypts into the intestinal lumen or whether the dilated crypts are a mucosal reaction due to another undetermined lesion that is responsible for alimentary tract protein loss is unknown. However, when large numbers of dilated intestinal crypts are present, they appear to be associated with PLE even if there are no other remarkable lesions in the intestinal mucosa. PMID- 10830545 TI - Contact factor deficiency in a German Shorthaired Pointer without clinical evidence of coagulopathy. PMID- 10830546 TI - Granulomatous tracheitis caused by Conidiobolus coronatus in a horse. PMID- 10830547 TI - An unusual neurological disorder in the Labrador retriever. PMID- 10830548 TI - Beyond the hematocrit and Po2: a symposium on teaching humanities in academic medical centers. PMID- 10830549 TI - Challenges in teaching ethics in medical schools. AB - Modern medical ethics has effected dramatic changes in medicine. Yet teaching medical ethics still presents many challenges. The main teaching methods used- inpatient ethics consultations, courses, and case conferences--have notable weaknesses. In addition, the attitudes and knowledge gaps of some learners may hamper these methods further. To encourage open discussion of the challenges, we outline our current approach to teaching medical ethics. We teach with the conviction that ethics instruction gives physicians vital knowledge not available from science. Our teaching addresses ethical issues directly relevant to residents and students, emphasizes a few important concepts, and nurtures learners' critical reasoning skills. Our teaching also tries to use scarce faculty time efficiently. However, we believe successful medical ethics teaching requires medical schools to commit significant material and moral support. We hope the discussion here encourages medical ethics teachers everywhere to describe the challenges they face and to collaborate on finding solutions. PMID- 10830550 TI - From Laennec to lobotomy: teaching medical history at academic medical centers. AB - Although clinicians without a sense of history may not be condemned to repeat the past, the historical record offers many informative lessons. For one thing, history demonstrates the changing nature of scientific knowledge; current understandings of health and disease may prove as ephemeral as earlier discarded theories. In addition, history reminds us that social and cultural factors influence how physicians diagnose and treat various medical conditions. When attempting to teach the history of medicine at academic medical centers, instructors should be innovative as opposed to comprehensive. Students and residents are likely to find recent historical issues to be more relevant, particularly when such material can be integrated into the existing curriculum. Provocative topics include depictions of medicine in old Hollywood films, the contributions made by famous physicians at one's own institution, and historical debates over controversial events, such as the Tuskegee syphilis study and the use of lobotomy in mental institutions in the 1950s. PMID- 10830551 TI - Reading, writing, and doctoring: literature and medicine. AB - Literature and medicine share an inherently enduring relationship. Doctors turn to literature--both its plots and its forms--to understand what occurs in their patients' lives, to increase their own narrative competence, to interpret accurately the texts of medicine, to develop empathy, and to deepen their capacities for reflection and self-knowledge. Together, these skills, attitudes, and bodies of knowledge contribute to the effective practice of medicine. Literature is now taught in almost three quarters of the medical schools in the United States. Different goals, agendas, and methods are appropriate at each developmental stage of a physician's training, from the premedical curriculum to the continuing education of a practicing physician. A vigorous and growing scholarship and body of experience is propelling the field of literature and medicine to understand all the more clearly how acts of reading and acts of writing might illuminate acts of doctoring. PMID- 10830552 TI - Seeing patients and life contexts: the visual arts in medical education. AB - In many ways, the practice of medicine has been a visual science from the time of the early Renaissance anatomists to the high-speed scanners of today. But images of patients and their anatomical parts do not necessarily lead to an understanding of their problems. Meaning must follow the sensory experience and be coupled with reflection. The visual arts, therefore, can be used to help physicians in training increase their observational and interpretive skills. Works by classic and contemporary artists can be used to increase awareness of the complex nature of human beings and their conditions, which lie beneath the appearances. In addition to painting, television, motion pictures, and printed media may also be used in classroom settings to educate. Medical schools that do not have accessible fine arts or humanities programs may form allegiances with local artists to increase communication and understanding between these disciplines. PMID- 10830553 TI - Teaching anthropology in the medical curriculum. AB - Practicing medicine well requires recognizing the breadth of human experience and attending to the psychological and sociocultural dimensions of patients as well as their physical needs. Central to the concerns of anthropology are the shared beliefs and values expressed in social practices and traditions that give meaning to everyday life. The relevance of anthropology for biomedical practice and research is grounded in the discipline's emphasis on contextual meaning and its unique strategies for data gathering. In this article, we briefly review the field of anthropology and the discipline of medical anthropology. We argue for incorporating anthropological concepts and methods in medical training, and summarize anthropology's role in medical education over the past century. Finally, we present ideas for including anthropology in the medical curriculum, proposing curricular goals and content, and teaching settings and techniques. An anthropological orientation can foster trainee self-awareness, help trainees prepare for the diverse perspectives they will encounter in our pluralistic society, and facilitate critical analysis of biomedicine and its systems of care. PMID- 10830554 TI - The future catches up: a medical school curriculum in health economics. AB - Medical school curricula for too long have ignored the obvious and important changes in the economics and delivery of health care in the United States. Medical students, who become the practicing physicians of the future, and their patients have suffered because of this academic malaise. Most new physicians are even more confused than their patients about how to navigate outpatient managed care, how to practice efficiently (after being taught that more is better), and how to uphold their sacred trust with their patients in the context of institutionally based medicine. After summarizing relevant historical events, we discuss how we hope to begin making up for lost time by tackling the issue of necessary curriculum change at the University of Pennsylvania Health System. PMID- 10830555 TI - Interventional revascularization of left main coronary artery stenosis with new devices: two cases of "unprotected" left main stenosis treated with atherectomy and stenting. PMID- 10830556 TI - Focal segmental glomerulosclerosis in African Americans: effects of steroids and angiotensin converting enzyme inhibitors. AB - BACKGROUND: Focal segmental glomerulosclerosis (FSGS) is a common primary glomerulopathy in African Americans. In this report, we present data on 40 African American patients with FSGS from our medical center. METHODS: Patients were identified from a review of all charts seen in our conservative management renal clinic in 1996, a review of renal biopsy rolls (1994-1998), and a review of patients entering the end-stage renal disease (ESRD) program with a primary diagnosis of FSGS (1993- 1997). Charts were reviewed for demographic, biopsy, and treatment data. Patients who were observed for at least 4 months (range, 4-125 months) were included. ESRD was used as the primary endpoint (n = 12). Data were analyzed using univariate and multivariate Cox hazards and Kaplan-Meier survival analysis. Twenty-four patients were treated with angiotensin-converting enzyme (ACE) inhibitors. Similarly, 24 patients were treated with corticosteroids for a mean of 8.75 +/- 2.6 months and a total dose of 9.3 +/- 2.2 g. RESULTS: On univariate analysis, factors found to be significant determinants for reaching ESRD were the initial creatinine (P = 0.0001), interstitial fibrosis (P = 0.032), the percentage of globally sclerosed glomeruli (P = 0.0018), and the mean arterial blood pressure over the course of follow-up (P = 0.05). Neither the ACE inhibitors nor the corticosteroids had a significant impact on reaching ESRD. The patients reaching ESRD (n = 12) were analyzed separately. The mean time from biopsy to ESRD was 24.7 +/- 9.8 months. ACE inhibitors prolonged renal survival (P = 0.023), but steroids did not. Initial creatinine was the only factor found to be a significant determinant for ESRD. CONCLUSIONS: We conclude that FSGS is common in African Americans. Early diagnosis and blood pressure control are important, but the beneficial effects of steroids and ACE inhibitors in this population are still unclear. PMID- 10830558 TI - Dose-response profile of acarbose in older subjects with type 2 diabetes. AB - OBJECTIVE: To examine the dose-response relationship of acarbose, an alpha glucosidase inhibitor, in older subjects with type 2 diabetes. RESEARCH DESIGN: Fourteen subjects with type 2 diabetes who were over 65 years old were studied. Five subjects had been treated with diet alone and 9 were receiving a sulfonylurea. The subjects underwent a meal tolerance test in the presence of varying doses of acarbose (0, 25, 50, and 100 mg) on 4 occasions, each 1 week apart. The test meal was chosen to include food items commonly consumed during breakfast in the United States. The 483-kcal meal consisted of 51% of calories in the form of carbohydrates, 14% protein, and 35% fat. The serum glucose, insulin, and triglyceride levels were measured at 0, 1, and 2 hours after the meal. RESULTS: The postprandial hyperglycemic response to the test meal was significantly reduced with 25 mg of acarbose compared with baseline values. Increasing doses of acarbose to 50 or 100 mg had no significant additional ameliorating effects on postprandial hyperglycemia. Postprandial insulin or triglyceride levels were not significantly altered with single dose acarbose treatment. CONCLUSIONS: It is concluded that the acute efficacy of acarbose is near maximal at 25 mg when the meal size does not exceed 483 kcal and contains only 61 gm of carbohydrates. PMID- 10830557 TI - Restricting medicaid payments for transportation: effects on inner-city patients' health care. AB - BACKGROUND: Prior approval programs have been used to control spiraling costs of Medicaid, but they are rarely formally assessed. We evaluated the effect of a change in Indiana Medicaid's policy (effective October 1, 1993) requiring prior approval to pay transportation costs. METHODS: We performed a historical cohort study comparing the health care utilization of Medicaid patients during the first 6 months of 1993 versus the first 6 months of 1994. Subjects included all Medicaid patients who visited any inpatient or outpatient site affiliated with an inner-city public hospital in the first 6 months of 1993 (N = 23,015) and 1994 (N = 23,707). RESULTS: These Medicaid patients made 82,961 visits in the first 6 months of 1993 and 79,809 visits in the first 6 months of 1994. Visits to hospital-based primary care clinics declined 16% (P < 0.001), which was partially offset by a 7% increase in visits to neighborhood health centers (P < or = 0.001). Emergency and urgent visits fell by 8%; visits for medication refills fell by 18% (P < 0.001 for each). Hospitalizations increased slightly in 1994, with no change in the number of inpatient days. There was no change in inpatient or outpatient nontransportation charges. There were no systematic reductions in selected aspects of preventive care. However, there were fewer emergency and urgent visits among patients with reactive airway disease. CONCLUSIONS: Requiring prior approval for transportation was associated with reductions in visits for primary care visits and refilling prescriptions without measurable short-term effects on charges or selected clinical parameters. Neighborhood health centers partially ameliorated the decline in primary care visits. PMID- 10830559 TI - Convulsions induced by metronidazole treatment for Clostridium difficile associated disease in chronic renal failure. AB - Clostridium difficile-related diarrhea and colitis are common health problems, especially in elderly, frail hospitalized patients. The drug of choice is metronidazole, which can be associated, in long or high doses, with neurotoxic side effects. We report convulsions induced by short-term metronidazole therapy used in conventional doses for Clostridium difficile colitis in an elderly patient with chronic renal failure. PMID- 10830560 TI - Idiopathic adrenal hemorrhage. AB - A case of idiopathic adrenal hemorrhage is reported. A 76-year-old woman exhibited a left adrenal tumor, 3 cm in diameter, on abdominal computed tomography. The patient was receiving aspirin medication for atrial fibrillation. There was no evidence of increased adrenal hormones. The mass enlarged to 6 cm in diameter within 18 months, and malignancy was suspected. The mass was diagnosed as adrenal hematoma by operative findings. PMID- 10830561 TI - The relationship between social support and major depression: cross-sectional, longitudinal, and genetic perspectives. AB - Although social support (SS) is associated with risk for major depression (MD), we are uncertain of the extent to which a) low SS increases risk for MD, b) MD lowers SS, or c) both variables reflect a common genetic liability. Using two waves of interview data on female twin pairs from a population-based registry, we examine the cross-sectional and longitudinal association of eight dimensions of perceived SS and MD. Risk for MD in the last year was inversely associated with supportive spouse and relative relationships, and directly associated with problems in these relationships (e.g., too many demands, criticism, tension, and disagreements). Significant cross-time associations were seen only for spousal variables. The history of MD in one twin significantly predicted low relative and spouse support, and relative and friend problems, in her co-twin (MZ > DZ pairs). The relationship between SS and MD in women is complex and due to at least the three separate mechanisms outlined above that operate to varying degrees in different dimensions of SS. PMID- 10830562 TI - Deployment stressors and a chronic multisymptom illness among Gulf War veterans. AB - Unusual health problems have been reported by Gulf War (GW) veterans, but no single etiology has been linked to these illnesses. This study was conducted to determine the association between self-reported GW deployment stressors and an illness defined by a combination of fatigue, mood-cognition, and musculoskeletal symptoms. A total of 1002 GW veterans from this cross-sectional survey of four Air Force units completed a self-administered questionnaire that asked about symptoms, demographic and military characteristics, and stressors during deployment. Severe and mild-moderate illness was positively associated with self reports of pyridostigmine bromide use, insect repellent use and belief in a threat from biological or chemical weapons. Injuries requiring medical attention were only associated with severe illness. These results suggest a link between self-reported chemical, emotional, and physical exposures, and GW veterans' illness. Further research is needed to determine physiological and psychological mechanisms through which such stressors could have contributed to this symptom complex. PMID- 10830563 TI - Peritraumatic dissociation following motor vehicle accidents: relationship to prior trauma and prior major depression. AB - Individuals who dissociate at the time of a traumatic event (peritraumatic dissociation) are more likely to develop acute and chronic posttraumatic stress disorder (PTSD). However, little is known about who is at risk of peritraumatic dissociation. Motor vehicle accident subjects (N = 122) were systematically recruited and followed over 12 months. We used the Structured Clinical Interview for DSM-III-R (SCID) and the Peritraumatic Dissociative Experiences Questionnaire Rater Version (PDEQ-RV). Younger subjects were more likely to experience peritraumatic dissociation as were white versus nonwhites, and single versus married subjects. Younger subjects reported a greater number of peritraumatic dissociative symptoms as did subjects with an injured passenger. After adjusting for age and passenger injury, prior major depression was significantly related to more peritraumatic dissociative symptoms. An interaction of age and prior major depression indicated that those who were younger and reported a history of major depression had the greatest number of peritraumatic dissociative symptoms. PMID- 10830564 TI - Coping in dieting and eating disorders: a population-based study. AB - The use of different coping strategies, measured by the Ways of Coping Questionnaire was investigated among 1157 women (18 to 30 years), randomly selected from the general population of Sweden as part of a longitudinal study. Subjects were clustered into five groups: subjects with past or current eating disorders (ED), and subjects with no ED but with past, current, or no history of dieting. Subjects with past or current ED reported significantly higher levels of escape avoidance and lower levels of seeking social support and purposeful problem solving compared with subjects with neither ED nor dieting. These group comparisons were then reanalyzed with sum of depressive symptoms as a covariate in covariate analyses. The only significant difference between the groups concerned the use of escape avoidance. The significant differences in the use of escape-avoidance strategies may motivate more extensive training in coping in the context of prevention and treatment of ED and maladaptive dieting. PMID- 10830565 TI - Stress coping strategy in Japanese patients with eating disorders: relationship with bulimic and impulsive behaviors. AB - Although maladaptive coping strategies in eating disorder patients have been reported, the relationship between impulsivity and coping strategy has not previously been studied. Subjects consisted of 43 patients with anorexia nervosa restricting type (AN-R), 42 patients with anorexia nervosa binge eating/purging type (AN-BP), 71 patients with bulimia nervosa purging type (BN), and 97 controls. The Coping Inventory for Stressful Situations was used to evaluate coping strategies. Only AN-BP patients had a significantly lower task oriented coping score than controls, and AN-R and BN patient groups used significantly less social diversion-avoidance coping strategies than controls. Emotion-oriented coping scores of AN-BP and BN patients were significantly higher than those of controls. In addition, impulsive BN patients had significantly higher emotional coping scores than less impulsive BN patients. These results suggest that maladaptive coping strategies may be a perpetuating factor even for impulsive patients and emphasizing a change in maladaptive coping strategies may be a useful treatment strategy even for highly impulsive patients. PMID- 10830566 TI - Prognostic significance of antisocial personality disorder in cocaine-dependent patients entering continuing care. AB - This study examined the relationship of antisocial personality disorder (APD) to response to continuing care treatments in a sample of cocaine-dependent patients. Patients (N = 127) were randomly assigned to 20-week standard group or individualized relapse prevention continuing care interventions after the completion of an initial treatment episode and followed up at 3, 6, and 12 months. APD and non-APD patients did not differ on retention in continuing care, substance use outcomes, social function outcomes, or experiences before or during cocaine relapse episodes. A diagnosis of APD was also not a predictor of differential response to the two continuing care interventions in the study. However, APD patients had worse medical and psychiatric problem severity than non APD patients at entrance to continuing care and during follow-up. These results suggest that cocaine patients with APD who are in the continuing care phase of outpatient rehabilitation might benefit from additional medical and psychiatric treatment services. PMID- 10830567 TI - Comparison of self-ratings and therapist ratings of outpatients' psychosocial status. AB - In psychiatric treatment, differences between therapists' observer ratings and patients' self-ratings are well known. We studied these differences in a sample of chronically mentally ill outpatients. The results show that the patients rated their psychosocial status significantly better than their therapists. By means of multiple regression analysis, we designed a model to explain the specific differences. By placing more emphasis on leisure activities and less emphasis on addictive behavior, compliance, and psychopathology, therapists might predict global ratings given by patients more accurately. This model helps therapists obtain a better understanding of their patients. PMID- 10830568 TI - The Spanish version of the quality of life index: presentation and validation. AB - Latino or Spanish-speaking individuals constitute a substantial and growing population in the United States, in addition to their general presence, with cultural variations, throughout Latin America and the Iberian Peninsula. To respond to the needs of this population, a Spanish version of the Quality of Life Index (QLI-Sp) was developed. The QLI, in its various language versions, is a concise instrument for comprehensive, culture-informed, and self-rated assessment of health-rated quality of life. It is composed of 10 dimensions collated from the international literature, including aspects ranging from physical well-being to spiritual fulfillment, as well as a global perception of quality of life. Each item is to be rated on a 10-point line by Latino subjects according to their culture-informed understanding of that concept. The study samples included 60 Latino psychiatric patients (20 outpatient, 20 inpatient, and 20 partial hospitalization) and 20 Latino actively working hospital professionals. Mean time of completion was 2.4 minutes among health professionals and 3.6 minutes among patients. The vast majority of respondents (72% of patients and 1000% of professionals) judged the instrument as easy to use. The test-retest reliability correlation coefficient of the QLI-Sp mean score was .89. The discriminant validity of the QLI-Sp was documented by the highly significant difference obtained between the mean scores of the two samples selected to represent quite different levels of quality of life. PMID- 10830569 TI - Defense styles and posttraumatic stress symptoms. PMID- 10830570 TI - Patient self-report in the assessment of panic disorder: comparison with interview-derived clinician ratings. PMID- 10830571 TI - Hitler's diagnosis. PMID- 10830572 TI - 'Mediterranean diet' and DNA adducts. PMID- 10830573 TI - Biological mechanisms in breast cancer invasiveness: relevance to preventive interventions. AB - Migration studies suggest that the high incidence of postmenopausal breast cancer in Western women is related mainly to epigenetic factors. Progression from ductal carcinoma in situ (DCIS) to invasive ductal carcinoma (IDC) also appears to involve environmental rather than genetic factors, and a role has been postulated for metabolic-endocrine changes related to the Western lifestyle. Protein kinase C (PKC) is important in cell signal transduction, and laboratory studies show that PKC stimulates the activities of urokinase plasminogen activator, matrix metalloproteinases and cell adhesion molecules, all of which are known to increase invasiveness in human mammary cancer cell lines. In rodents, the activity of PKC in tissue cells is enhanced by insulin, and PKC isoenzymes have been shown to stimulate the development of hyperinsulinaemic insulin resistance in rodents. Clinically, hyperinsulinaemia and the concomitant increase in circulating levels of free oestradiol and bioactive insulin-like growth factor 1 (IGF1) are each confirmed markers of high risk for breast cancer in women. Lesions of DCIS show evidence of regression with mammary involution, but it is postulated that this may be opposed by the concomitants of hyperinsulinaemic insulin resistance. The prevalence of the latter is increasing in Western populations, and a combination of high IGF1 and low IGF-binding protein 3 concentrations has been associated with the presence of DCIS lesions in premenopausal women. Measures that enhance insulin sensitivity in such women may reduce the risk of progression in DCIS lesions, and a clinical trial is proposed to test the hypothesis. PMID- 10830574 TI - p53 gene mutation and protein expression in operable non-small cell lung cancer in Poland. AB - We investigated the association of p53 abnormalities (gene mutations by DNA sequencing and protein over-expression by immunostaining) with clinical data and prognosis in 74 patients with resected non-small cell lung cancer (NSCLC). DNA analysis of exons 5-8 of the p53 gene showed 34 mutations in 74 resected primary NSCLC (45.9%). Immunohistochemical study of the p53 protein revealed that 41 of 74 (55.4%) samples had positive staining. We found strong agreement between the results of the p53 protein expression test (p53-PE) and the p53 gene mutation test (p53-M) (Cohen's kappa = 0.65, 95% CI 0.48-0.82). Joint distribution of the results (analysed using the bivariate Dale model) was mainly influenced by, histological type of tumour. A positive result for the p53-PE test significantly increased (estimated odds ratio 84.5; 95% CI 8.89-803.03) the odds of observing a positive result in the p53-M test. In the univariate analysis (log rank test), positive results in the p53-M test and the p53-PE test were significantly associated with overall survival (P < 0.001 and P = 0.005, respectively). In the multivariate analysis (Cox's proportional hazard model), a positive result for the p53-M test significantly increased relative risk for overall survival (RR 9.56; 95% CI 2.62-34.87; P < 0.001). When the result of the p53-M test was accounted for, a positive result for the p53-PE test did not offer any additional prognostic information due to the strong dependence of results of the tests. However, when the result of the p53-M test was removed from the model, a positive result for the p53-PE test became a significant unfavourable prognostic factor (P = 0.009). We conclude that p53 gene mutation and protein expression analyses are in a strong agreement. Joint distribution of the results depends mainly on histological type of tumour. When considered separately, both tests are unfavourable prognostic factors in NSCLC. When the result of the p53-M test is taken into account, the p53-PE test does not offer any additional prognostic information. PMID- 10830575 TI - Diet and stomach cancer: a case-control study in South India. AB - A prospective case-control study was conducted in Trivandrum, India, to evaluate the dietary risk factors for stomach cancer. One hundred and ninety-four patients with stomach cancer registered at the Regional Cancer Centre (RCC), Trivandrum, Kerala, India, during the period 1988-1991 were considered as cases. A minimum of one control (n = 305), matched for age (+/- 5 years), sex, religion and residential area was selected from the visitors to RCC during the same period. Interviews were carried out using a predetermined structured food frequency questionnaire. The information collected also included socio-demographic/economic background, tobacco chewing, tobacco smoking and alcohol habits. Data were analysed using a multiple logistic regression model. Odds ratios for all dietary variables were estimated. Increased risks were observed with higher consumption of rice (OR 3.9; 95% CI 1.6-10.0). Risk was high for those consuming spicy food (OR 2.3; 95% CI 1.1-5.0), high consumption of chilli (OR 7.4; 95% CI 4.0-13.5) and consumption of high-temperature food (OR 7.0; 95% CI 3.7-12.9). On multivariate analysis, high consumption of rice, high consumption of chilli and consumption of high-temperature food were found to be independent risk factors. PMID- 10830576 TI - Sex differences in colorectal cancer mortality in Europe, 1955-1996. AB - Colorectal cancer is the leading cancer in non-smokers in Western countries, and over the last decades its trends have been generally more favourable for women than for men. Possible explanations of the sex differentials in colorectal cancer relate to different exposure to exogenous hormones and to other risk factors including diet, physical activity and alcohol drinking. The objective of this investigation was to systematically analyse the trends in colorectal cancer mortality sex ratios in major European countries over the last four decades. Trends in death certification rates from colorectal cancer over the period 1955 1996 were analysed for 20 European countries (excluding the former Soviet Union and a few of the smaller countries). In all countries, the mortality sex ratios (M/F) were around or slightly above unity in the 1950s, and systematically increased to approach 1.5 in the 1990s. The extent of the rises varied across countries, ranging between + 0.8% in Germany, + 9.7% in Sweden, and + 12.1% in Denmark (the lowest increases) to + 65.3% in Spain, + 56.2% in Portugal, and + 50.4% in Hungary (the highest ones). Mortality sex ratios in Europe show more favourable trends for females, which may be attributable, in part, to the introduction of exogenous hormones in the late 1950s and 1960s, and, in part, to differential sex exposure to major environmental risk factors. PMID- 10830577 TI - Parity and mammographic breast density in relation to breast cancer risk: indication of interaction. AB - We examined whether the harmful influence of nulliparity on breast cancer risk could be mediated by high mammographic density. Another possibility is that mammographic density and nulliparity act independently or perhaps synergistically on breast cancer risk. Our study population consisted of 129 cases and 517 controls who had been participants in the Nijmegen breast cancer screening programme for 10 years. Breast density was classified with a fully automated technique on digitized mammograms from the screening examination 10 years before diagnosis. Classification was based on the proportion of the breast that was composed of high density: < 5%, 5-25% or > 25%. Data on parity and potential confounders were obtained using a questionnaire, administered at the same examination. We found that nulliparae with low breast density (< 5%) were not at increased risk compared to parous women with low density: OR 1.1 (95% CI 0.2 5.8). Parous women with < 5% density formed the reference category throughout all analyses. The risks for parous women with 5-25% or > 25% density were 2.7 (95% CI 1.3-5.6) and 3.6 (95% CI 1.7-7.7) fold increased, respectively. However, when both factors were present (nulliparity and > or = 5% density), breast cancer risk was 7.1 times higher (95% CI 3.2-15.9). This could indicate that nulliparity and high breast density might work synergistically and that breast density is not just an explanatory factor in the influence of nulliparity on breast cancer risk. It is hypothesized that high breast density (reflecting fibro-glandular tissue with increased epithelial cell proliferation) is more susceptible to carcinogenic effects in the undifferentiated epithelial breast tissue of nulliparae than in the differentiated tissue of parous women. Since there were few data, no firm conclusions can be drawn. If these findings can be confirmed in a larger study population, however, they may have important implications for the prevention and early detection of breast cancer. PMID- 10830578 TI - p53 codon 72 polymorphism and risk of intra-epithelial and invasive cervical neoplasia in Greek women. AB - In 1998, Storey and co-workers suggested that individuals homozygous for arginine (Arg) at codon 72 of the p53 gene are about seven times more susceptible to human papillomavirus (HPV)-related carcinogenesis than heterozygotes. Since then, several studies from Northern Europe, Japan and the USA have failed to demonstrate a similar correlation. By contrast, a study in Brazil as well as one recent study in Italian and Swedish populations showed strong positive associations. We examined the frequency of p53 codon 72 polymorphism in samples from both invasive and intra-epithelial cervical neoplasias (CIN), and compared them with samples from healthy controls. All 88 samples came from women with a Greek ethnic background. Tissue specimens were collected from archival material with histologically diagnosed low-grade CIN (LGCIN), high-grade CIN (HGCIN) or cervical cancer (CxCa). As a control, we used cellular material newly collected by cytobrush from the cervices of 30 healthy women with normal cytological and colposcopical examinations. p53 Arg homozygosity (Arg/Arg) alone was associated with four-, six- or eight-fold increased risks for LGCIN, HGCIN or invasive cancer, respectively. The frequency of the p53Arg/Arg genotype and of the proline (Pro) allele showed significant linear trends according to the degree of severity of the lesion (P = 0.0007 and P = 0.0009, respectively). Exclusion of the ten HPV16/18-negative cases did not substantially alter the Arg/Arg frequency among the groups nor the significant linear trend. Our results confirm the initial findings of Storey and co-workers, as well as the data of the Brazilian and the recent European study, but do not accord with those of the other aforementioned studies. Variations in ethnic background, laboratory performance, verification of the HPV status, definition of controls, and sample size are the most plausible explanations for this controversy. In all our samples, the distribution of the p53 alleles fits the Hardy-Weinberg equilibrium and the 0.48 frequency of the Pro allele in our controls accords well with the percentages previously reported for different ethnic groups as characteristic of the assumed north-south cline. Some authors assert that the discrepancy in the results could not be attributed to differences in the methods; however, the Brazilian study emphasized the effect of inter-laboratory variation in detecting the association between p53 polymorphism and cervical cancer. Regarding the control group, our samples were only from women with a cytologically and colposcopically benign cervical epithelium. We think that simply choosing 'normal volunteers' for collecting control DNA blood samples without knowing the status of their cervical epithelium is indeed a possible source of bias. Finally, it is very unlikely that loss of heterozygosity at the p53 locus could be a factor interfering with the allelotype distribution. Our present small study results, which suggest a biologically relevant association, provide strong evidence that homozygous arginine at codon 72 of p53 may confer a higher susceptibility to HPV-associated intra-epithelial and invasive cervical neoplasia. PMID- 10830579 TI - Fraction of prostate cancer incidence attributed to diet in Athens, Greece. AB - Diet appears to be a major determinant in the incidence of prostate cancer. In a case-control study conducted in Athens, Greece, we found that dairy products, butter and seed oils were positively associated with risk of prostate cancer, whereas cooked and raw tomatoes were inversely associated. We utilized the data from this study to calculate the population attributable fractions under alternative assumptions of feasible dietary changes. For each subject, a dietary score was calculated and categorized into approximately quintiles, representing increasing levels of prostate cancer risk as a function of the intake of the five discriminatory food groups or items. Population attributable fractions in terms of this dietary score were calculated taking into account multivariate adjustment. We observed that, if all individuals were shifted to the baseline category, the incidence of prostate cancer in this study population would be reduced by 41% (95% confidence interval 23-59%). However, if all individuals were shifted to the adjacent lower risk quintile, the expected incidence reduction would be a more modest 19%. The incidence of prostate cancer in Greece could be reduced by about two-fifths if the population increased the consumption of tomatoes and reduced the intake of dairy products, and substituted olive oil for other added lipids. PMID- 10830580 TI - Tea and other beverage consumption and prostate cancer risk: a Canadian retrospective cohort study. AB - Using participants in the 1970-1972 Nutrition Canada Survey (NCS), a retrospective cohort study was conducted to assess the relationship between tea, as well as coffee, cola and alcohol, and the risk of developing prostate cancer. The mortality and cancer experience of male NCS participants aged 50-84 years was determined up to 31 December 1993. Among the 3400 survey participants included in the study, 145 developed prostate cancer. No association was observed between tea (predominantly black tea) intake and prostate cancer. Subjects who drank more than 500 ml of tea per day experienced virtually the same risk as those who reported no tea consumption (rate ratio (RR) 1.02, 95% confidence interval (CI) 0.62-1.65). Compared to those who reported no coffee drinking, men who averaged more than 250-ml per day experienced a 40% increase in risk (95% CI 0.84-2.32). Cola consumption was not associated with an increased risk of prostate cancer. Total alcohol consumption was not related to subsequent development of prostate cancer, although very moderate consumption of wine (< 10 g per day), relative to no consumption, showed an RR of 1.48 (95% CI 1.05-2.09). These data do not support an association between consumption of tea and prostate cancer risk. PMID- 10830581 TI - Relationship between anal occult blood and hepatobiliary pancreatic carcinoma screening. AB - Cotton swab anal smear instead of stool occult blood test can not only be used as a mass screening method for colorectal cancer, but is an auxiliary way to screen hepatobiliary pancreatic carcinoma. Two hundred and twenty-three cases of hepatobiliary pancreatic carcinoma patients received anal occult blood test, of which 121 were positive (54.3%). The screening test was performed in 14,645 healthy people aged 40-60 years; 511 persons (3.4%) were positive for anal blood. Among these, six cases of digestive tract cancer were found (one oesophageal carcinoma, one gastric cardia carcinoma, two gastric carcinomas, two liver carcinomas). This suggests that when anal occult blood is positive, the patient should be scanned by ultrasonic means to rule out hepatobiliary pancreatic tumour. PMID- 10830582 TI - Sunlamps and sunbeds and the risk of cutaneous melanoma. Italian Group for Epidemiological Research in Dermatology. PMID- 10830583 TI - The position of the opposite flat applicator changes the SAR and thermal distributions of the RF capacitive intracavitary hyperthermia. AB - The variations of the specific absorption rate (SAR) and thermal distribution in the JSHO QA phantoms were investigated by radiofrequency (RF) capacitive intracavitary hyperthermia (ICHT) applicator (AP-T01, Omron Electric Co., Kyoto, Japan) with the changing position of the opposite flat applicator (15-cm in diameter). The thermal distribution was observed with the thermographic camera and the normalized SAR distribution was calculated with the thermal data that were measured with the thermocouple thermometers. The SAR and thermal distributions of AP-T01 significantly varied with the position of the opposite flat applicator. The slope of the normalized SAR became gradual towards the side of the flat applicator. During the operating of a high flow rate (1500 ml/min) cooling system, the region between AP-T01 and the flat applicator was widely and rather homogeneously heated, except the hot spot around the end of AP-T01. This hot spot may be due to the imbalance of cooling of AP-T01 and the warming-up of the electrode. These results suggest that the RF capacitive ICHT using AP-T01 may be clinically effective on the deep-seated tumours in the direction of either the end wall, such as cervical cancers, or the upper wall, such as prostatic cancers and the mediastinal metastatic lymph nodes, if an improvement of the cooling system is achieved. PMID- 10830584 TI - The size and distance of the opposite flat applicator change the SAR and thermal distributions of RF capacitive intracavitary hyperthermia. AB - The variations of the specific absorption rate (SAR) and thermal distributions in the JSHO QA phantom were investigated by using the radiofrequency (RF) capacitive intracavitary hyperthermia (ICHT) applicator (AP-T01, Omron Electric Co., Kyoto, Japan) and the opposite flat applicators of different sizes (AP-75E: 7.5 cm in diameter, AP-100E: 10 cm in diameter, and AP-150E: 15 cm in diameter). The influences of the distance between both applicators were also investigated. Heating of the region between both applicators became weaker with the increase in size of the opposite flat applicator, and it became stronger with the decrease of the distance between both applicators. Heating near the flat applicator became weaker with the increase in size of the flat applicators, and it showed no apparent difference with the increase of the distance between both applicators. The normalized SAR values between AP-T01 and the opposite flat applicator became smaller and its slope became steeper with the increase in size of the opposite flat applicator and in the distance between both applicators. These results suggest that the variability of the specific absorption rate (SAR) and thermal distributions of the region between both applicators may show the potentiality of usefulness for heating the tumours of various sizes, shapes and location. PMID- 10830585 TI - Dielectric-loaded coaxial-slot antenna for interstitial microwave hyperthermia: longitudinal control of heating patterns. AB - In this paper, the microwave interstitial antenna with the dielectric load in part near the tip is introduced to realize the tip-heating and to improve the dependence of the heating patterns on the insertion depth. Numerical simulations using the Finite Difference Time Domain (FDTD) method have been conducted at the frequency of 915 MHz for four different configurations of the coaxial-slot antenna inserted into a catheter: the media between the antenna and the catheter are (a) no, (b) a thin air layer, (c) a thin dielectric layer, and (d) a thin air layer and a dielectric load in part near the tip. The diameter of the antenna including the catheter is sufficiently small for minimally invasive therapy. Comparison of the SARs for the four configurations makes it clear that the dielectric-loaded antenna can realize the best tip-heating and suppress the hot spot near the surface of the human body. Dependence of the SAR distributions on the insertion depth of the antenna has also been examined. It is found from the investigation that the dielectric-loaded antenna has little dependence on the insertion depth. PMID- 10830586 TI - Molecular chaperone function of mammalian Hsp70 and Hsp40--a review. AB - Virtually all organisms respond to up-shifts in temperature (heat shock) by synthesizing a set of proteins called heat shock proteins (HSPs). The HSPs are induced not only by heat shock but also by various other environmental stresses. Induction of HSPs is regulated by the trans-acting heat shock factors (HSFs) and cis-acting heat shock element (HSE) present at the promoter region of each heat shock gene. Usually, HSPs are also expressed constitutively at normal growth temperatures and have basic and indispensable functions in the life cycle of proteins as molecular chaperones, as well as playing a role in protecting cells from the deleterious stresses. Molecular chaperones are able to inhibit the aggregation of partially denatured proteins and refold them using the energy of ATP. Recently, there are expectations for the use of molecular chaperones for the protection against and therapeutic treatment of inherited diseases caused by protein misfolding. In this review, the focus will be on the mammalian Hsp40, a homologue of bacterial DnaJ heat shock protein, and the beneficial functions of molecular chaperones. PMID- 10830587 TI - The augmentative effect of repeated heat shock preconditioning on the production of heat shock protein 72 and on ischemic tolerance in rat liver tissue. AB - OBJECTIVE: Heat shock pretreatment induces heat shock protein (HSP)72 strongly in rat livers and provides the tolerance against subsequent ischemia-reperfusion injury. In this study, the effects of repeated heat shock pretreatment on the production of HSP72 in rat livers and on subsequent ischemic tolerance were investigated. METHODS: Rats pretreated with repeated heat shock were compared with those that received a single heat shock pretreatment. The production of HSP72 was analysed using Western-blotting and densitometer. At 48 h after heat shock pretreatment, all rats were subjected to warm liver ischemia for 30 or 45 min and then reperfused. Survival rate of the animals and liver functions during reperfusion were analysed. RESULTS: The production of HSP72 increased in the repeated heat shock group more than in the single heat shock group. Although there were no significant differences in animal survival or in liver functions after a 30-min ischemia between the single heat shock group and the repeated heat shock group, animal survival and liver functions after a 45-min ischemia were significantly better in the repeated heat shock group. CONCLUSION: In rats, repetition of heat shock pretreatment augmented the production of HSP72 in liver tissue and protected the liver from ischemia-reperfusion injury. PMID- 10830588 TI - The combined effect against colon-26 cells of heat treatment and immunization with heat treated colon-26 tumour cell extract. AB - Cancer vaccines represent a promising new strategy for immunotherapy against cancer, but their effects are insufficiently understood. The effect of heat treatment against mouse colon adenocarcinoma cell line (colon-26), and combined effects of heat treatment and immunizing host animals with heat treated colon-26 cell extracts were investigated. Heat treatment of colon-26 cells induced heat shock protein 70 (HSP70), but not other HSP. Immunization of BALB/cJ mice with heat treated colon-26 cell extract, which was enriched in HSP70, elicited antitumour immunity against subcutaneously injected colon-26 cells. Furthermore, combination therapy of heat treatment and immunization with heat treated colon-26 cell extract significantly reduced tumour volumes compared with heat treatment alone. Similar immunization enhanced the cytotoxic activity of mouse splenic lymphocytes against untreated and heat treated colon-26 cells in an in vitro assay, as well as against heat treated allogenic mouse lymphoma cell line (YAC 1). These findings suggest possible usefulness of heat treated cancer cell extract as a cancer vaccine, especially if given in combination with hyperthermia. PMID- 10830589 TI - Granulocyte-colony stimulating factor enhances anti-tumour effect of hyperthermia. AB - The combined effect of granulocyte-colony stimulating factor (GCSF) and hyperthermia in the treatment of experimental tumours was studied to examine the possible involvement of activated granulocytes in the antitumour effect of hyperthermia. Two weeks after transplantation of SCC VII cells (1 x 10(5)) into the instep of the left leg of C3H/HeJ male mice, the mice were given subcutaneous injections of GCSF (0.2 mg/kg) for 4 days. On day 4, hyperthermia was applied locally at 43 degrees C for 40 min. Hyperthermia inhibited the tumour growth, and this effect was enhanced by pre-treating the animals with GCSF. The numbers of circulating neutrophils in control and GCSF-treated mice were 2728 +/- 517/microl and 3124 +/- 194/microl, respectively (p = 0.53). Hyperthermia increased the number of neutrophils to 4409 +/- 700/microl (p < 0.05). Hyperthermia combined with GCSF significantly increased the number of netrophils to 5479 +/- 691/microl (p < 0.01). Chemiluminescence analysis using L-012 revealed that GCSF enhanced the generation of reactive oxygen species by about 10-fold. Glutathione contents in tumours 24 h after hyperthermia decreased by about 50% in both the hyperthermia groups with or without GCSF, as compared to those in the control. The GCSF-enhanced anti-tumour activity of hyperthermia was markedly inhibited by administration of a long-acting superoxide dismutase derivative (SM-SOD). These results suggest that GCSF activates the ability to generate active oxygen species by neutrophils and, thereby, enhances the anti-tumour effect of hyperthermia. PMID- 10830590 TI - WBH as a method of heating inaccessible tumours uniformly. PMID- 10830591 TI - In-hospital mortality after transurethral prostatectomy in Victorian hospitals. PMID- 10830592 TI - A community study of lower urinary tract symptoms in older men in Sydney, Australia. AB - BACKGROUND: The aim of the present paper was to determine the prevalence, bother attributable to and self-reported management of uncomplicated lower urinary tract symptoms (LUTS) in men aged 40-80 years in Sydney, Australia. METHODS: A total of 340 randomly selected men aged 40-80 years (65% response rate) participated in a community-based study (computer-assisted telephone survey). RESULTS: Lower urinary tract symptoms are common: 54% of men needed to wake up at least once at night to urinate; 47% indicated they had terminal dribbling 'sometimes' or 'frequently'; 30% experienced urgency although few (4%) had urge incontinence; 21% experienced hesitancy; and 19% could retain urine in their bladder during the day for no more than 2 hours. Urinary symptoms correlated poorly with self-rated bother and there was no increase in age-specific prevalence of bothersome symptoms with increasing age. Only 37 (26%) men inconvenienced by urinary symptoms had seen a general practitioner (GP) about these symptoms in the last 5 years: of these, two-thirds had been referred to a urologist and half of these received surgical treatment. Independent predictors of attending a GP were increasing age (adjusted odds ratio (AOR) = 12.3; P = 0.0015); place of birth outside Australia (AOR = 3.8; P = 0.0036) and anxiety about prostate cancer (AOR = 2.6; P = 0.0318), but not the degree of worry due to urinary symptoms. CONCLUSION: Lower urinary tract symptoms are common in men, but their experiences of bother correlate weakly with symptoms and do not appear to influence referral and treatment. Public and professional campaigns might increase wider understanding of the benefits of surgery for bothersome symptoms, not symptoms per se. PMID- 10830593 TI - An evaluation of trauma team response in a major trauma hospital in 100 patients with predominantly minor injuries. AB - BACKGROUND: A prospective study of trauma team response and performance at a major trauma service was undertaken between June and September 1998. METHODS: Following activation of the trauma team, the timing of the trauma team's arrival, the subsequent early management of the patient, time to monitoring, X-ray investigation and procedures performed were documented. RESULTS: The study evaluated 100 activations, 76% male, mean age 32 years and 65% were due to road trauma. The team leader, airway doctor and surgical registrar were present on patient arrival in 96%, 90% and 76% of cases, respectively. The airway, procedure and scribe nurses were present on patient arrival in 77%, 97% and 95% of cases, respectively. The radiographer was present in 69% of cases. Comparison between normal and after-hour response revealed little difference in medical and radiographer response, but the after-hour nurse response was significantly worse (P < 0.001). The median time to achieve electrocardiogram monitoring, blood pressure reading, and oxygen saturation tracing was 3 (range: 1-13), 4 (range: 2 20) and 3 (range: 1-21) min, respectively. Intravenous cannulation, phlebotomy and dispatch of bloods occurred at median times of 5 (range: 2-22), 6 (range: 3 23) and 17 (range: 7-40) min. The median times for intubation, chest tube and splintage of fracture were 10 (range: 3-19), 10 (range: 6-14) and 26 (range: 19 55) min, respectively. CONCLUSIONS: The present study identified an excellent multidisciplinary trauma response and provides a template to improve performance in early trauma management. PMID- 10830594 TI - Blunt trauma to the spleen. AB - BACKGROUND: The management of splenic injury resulting from blunt trauma in adults is controversial, with an increasing trend towards non-operative management and conservation of the spleen. A retrospective review was performed on adult patients treated in a single institution for splenic injury resulting from blunt trauma in an attempt to identify factors important in selecting an appropriate management option and predicting the success of that option. METHODS: Associated injuries (standardized using Injury Severity Scores), clinical signs at presentation, computed tomographic grading of splenic injury, and transfusion requirements were documented. Statistical analysis was performed using non parametric Mann-Whitney, Chi-squared, Kolmogorov-Smirnov and multivariate logistic regression tests. RESULTS: Eighty-five patients were identified. Non operative management was used on 39 patients, splenic conservation on 14 patients, and splenectomy on 32 patients. The mean Injury Severity Score was significantly lower in the non-operative group. Computed tomographic grading of the splenic injury was not found to correlate well with intraoperative findings. Transfusion requirements were lower in the non-operative group. Non-operative management failed in four patients; two had continued splenic bleeding, and two required surgery for other intra-abdominal injuries. Overall mortality was 7%. There was one death in the splenic conservation group, unrelated to the splenic injury, and two patients required a second laparotomy and splenectomy for persistent splenic bleeding. There were five deaths in the splenectomy group, only one of which was related to the splenic surgery. CONCLUSION: Management of blunt splenic injury remains controversial. The decision to pursue non-operative management rather than splenic conservation or splenectomy depends on the individual merits of each case. There is an increasing trend towards splenic conservation, particularly in younger, stable patients with single organ injury. PMID- 10830595 TI - Pelvic fracture pattern predicts pelvic arterial haemorrhage. AB - BACKGROUND: The association between pelvic fracture pattern and the need for pelvic arterial embolization remains controversial. To address this issue, a study of the experience at Auckland Hospital was undertaken. METHODS: Review was undertaken of a trauma database, blood bank database, patient records and pelvic radiographs. Of the 364 pelvic fracture patients admitted over a 4-year period, 76 were transfused with 6 or more units of blood in the first 24 h and these constitute the study population. RESULTS: Embolized patients were older (median age 42 vs 29.5 years; P < 0.05) and had a higher abbreviated injury score for the pelvic girdle (median 3 vs 2; P < 0.05) compared to non-embolized patients. Revised trauma score (median 7.69 vs 7.55), injury severity score (median 29 vs 30.5), morbidity (55 vs 39%) and mortality (45 vs 32.1%) rates did not differ significantly between embolized and non-embolized groups, respectively. The median blood transfusion requirement in the first 6 h from injury was 14 units in embolized and 8 units in non-embolized patients (P = 0.005). Embolization was required in 12 of 27 (44.4%) patients with fracture patterns indicative of major pelvic ligament disruption, whereas seven of 38 (18.4%) patients without these fracture patterns required embolization (P < 0.05). CONCLUSIONS: The need for pelvic embolization correlated with fracture patterns that indicated major ligament disruption, although the relationship was not sufficiently strong to warrant change to current indications for pelvic angiography. PMID- 10830596 TI - Improving basic surgical skills for final year medical students: the value of a rural weekend. AB - BACKGROUND: Hospitals employing medical graduates often express concern at the inexperience of new interns in basic surgical skills. In self assessment questionnaires, our senior medical students reported little clinical procedural experience. A practical skills workshop was staged in order to set learning goals for the final study year. This gave the students an opportunity to learn, revise and practice basic surgical techniques. METHOD: The Bruce Rock rural community sponsored a surgical camp at the beginning of the academic year. Ninety-five (80%) of the class registered at the workshop, which rotated them through teaching modules, with private study opportunities and the capacity to cater for varied skill levels. Eight teaching stations with multiple access points were provided, and ten mock trauma scenarios were staged to augment the learning process. RESULTS: The teaching weekend was rated by students on an evaluative entrance and exit questionnaire. Sixty-five (73%) students returned questionnaires. They recorded significant improvement (P < 0.05) in their ability to handle the teaching stations. All students had inserted intravenous lines in practice prior to the camp, so the rating change in intravenous line insertion ability was not statistically significant. CONCLUSIONS: The weekend retreat offers students a chance to focus on surgical skills, free from the pressures of a clinical setting or the classroom. The emphasis was on the value of practice and primary skills learning. Students endorsed the camp as relevant, practical and an enjoyable learning experience for basic surgical skills. PMID- 10830597 TI - Experience with the holmium laser as an endoscopic lithotrite. AB - INTRODUCTION: Holmium laser lithotripsy represents an additional option in the management of urinary tract calculi. We report the results of a cohort of patients with ureteric and bladder calculi treated with this modality. METHODS: Twenty-three patients underwent holmium laser lithotripsy to treat ureteric or bladder calculi. Power settings of 0.5 J to deliver 2.5-4.0 watts were used for the ureteric calculi, and up to 30 watts for the bladder calculi. RESULTS: The mean ureteric stone size was 7.3 mm (range 4-10 mm). One stone was upper ureteric, seven were mid-ureter and nine were lower-ureter. Eighteen of the 19 patients with ureteric calculi were free of stones 28 days postoperatively. The remaining patient was clear by 12 weeks. All patients with bladder calculi were completely cleared of stones. There were no intraoperative complications. DISCUSSION: Ureteric stone position can limit the use of extracorporeal shock wave lithotripsy due to imaging difficulties. Ureteric lithotripsy overcomes this problem. The holmium laser has proven to be safe and effective in clearing urinary stone burdens of a variety of sizes, sites and compositions in this cohort of patients. PMID- 10830598 TI - Clinical pathway management of total knee arthroplasty: a retrospective comparative study. AB - BACKGROUND: Clinical pathways facilitate the management of defined patient groups using interdisciplinary plans of care. The aim of the present study was to evaluate the effectiveness of a clinical pathway in improving a range of selected outcome measures in patients who have undergone total knee arthroplasty (TKA). METHODS: The present study was conducted at Queen Elizabeth Hospital, Adelaide. Using a retrospective comparative study design, 119 TKA patients who were managed on a clinical pathway from July 1997 to January 1998 (group 2) were compared with a retrospective group of 58 patients who underwent the same procedure from July 1996 to January 1997 (group 1) prior to the pathway's implementation. The following outcomes were measured: length of hospital stay; postoperative complications; readmissions and emergency service visits within 6 months of discharge; day of transfer to the convalescent unit; convalescent unit utilization and admission and discharge times. RESULTS: There was a significant reduction in the median length of stay in group 2 patients (9 vs 7 days; P < 0.0001). In addition there was a 66% increase in the proportion of patients in group 2 who were admitted on the day of surgery (P < 0.0001) and a 19.6% increase in the number of patients discharged within 8 postoperative days (P < 0.01). There were no significant differences between the groups with respect to the occurrence of postoperative complications. Although there was a trend toward a reduction in emergency service utilization and readmissions within 6 months of discharge for patients managed on the pathway, this was not significant. CONCLUSIONS: The development and implementation of a TKA clinical pathway resulted in a significant reduction in length of stay and improved streamlining of admission, discharge and transfer processes without adversely affecting patient outcomes. PMID- 10830599 TI - Phoenix date palm injuries: a review of injuries from the Phoenix date palm treated at the Starship Children's Hospital. AB - BACKGROUND: The purpose of this study was to review patients treated for penetrating Phoenix date palm injuries. Documentation of the clinical behaviour of these injuries was sought as they have different clinical outcomes to other foreign body injuries. METHODS: Data was collected via a retrospective review of patient files and imaging studies of all patients with foreign body injury admitted to the Orthopaedic Service of Starship Children's Hospital between 1992 and 1997. RESULTS: Of 250 patients admitted with foreign body injuries, 21 cases were identified as caused by the Phoenix date palm. The age range was 5 to 12 years. There was a median delay of 43 days between the time of injury and presentation to hospital. An average of 1.3 operations were performed on each patient. Ultrasound was used in 12 cases, foreign body was identified in 10 patients--a sensitivity of 83%. Intraoperative ultrasound was also found to be beneficial in three of these cases. CONCLUSIONS: Phoenix date palm injuries often have a protracted course. The initial diagnosis and treatment of such injuries is often delayed. Fracturing and fragmentation of the barbs may occur and multiple surgical procedures may be required to remove all of the foreign material. The Phoenix date palm barbs may cause a severe tissue reaction with sterile granulomatous synovitis and bony pseudotumour formation. A high level of suspicion on history taking and surgical exploration is needed. Total synovectomy for delayed joint or tendon sheath involvement may be required when fragmentation has occurred. Ultrasound has higher sensitivity for detection of thorn foreign bodies than magnetic resonance imaging. PMID- 10830600 TI - Squamous cell carcinoma of the lip: a retrospective review of the Peter MacCallum Cancer Institute experience 1979-88. AB - BACKGROUND: Squamous cell carcinoma (SCC) of the lower lip is a common malignancy in Australia. Surgical excision and/or radiotherapy are used in treatment, and are regarded as equally effective. METHODS: A retrospective review of 323 patients treated at the Peter MacCallum Cancer Institute with either surgical excision and/or radiotherapy, evaluated disease recurrence, cause-specific mortality, and the incidence of metachronous lesions. RESULTS: Recurrence-free survival at 10 years was estimated to be 92.5%, and cause-specific survival at 10 years was estimated to be 98.0%. Equivalent rates of local control were obtained with surgery and radiotherapy. Recurrence was related to tumour stage and differentiation. A high incidence of metachronous lesions was noted, 25 patients had a lesion prior to presentation and 33 patients developed second lip lesions during the study period. CONCLUSIONS: Squamous cell carcinoma of the lower lip is well treated with surgery or radiotherapy. The preferred treatment for most patients with SCC of the lower lip in the Australian population is surgical excision. This study has shown a significant incidence of metachronous lip neoplasia, except in those patients whose whole lip had been resurfaced. PMID- 10830601 TI - Calf augmentation. PMID- 10830603 TI - Laparoscopic Tenckhoff catheter insertion: a prospective study of a new technique. AB - BACKGROUND: This study describes a new technique of laparoscopic Tenckhoff catheter insertion. METHODS: During a 12-month period, 53 consecutive patients requiring insertion of peritoneal dialysis catheters were studied prospectively. A new technique of laparoscopic insertion was developed. RESULTS: Forty-eight patients out of 49 (98%) had successful insertion. Four patients were excluded with active peritonitis. There was an early complication rate of 8% (4/49) and a late complication rate of 14% (7/49). There was 12% overall failure of catheter with a mean follow up of 6 (1-13) months. CONCLUSION: This new laparoscopic technique maximizes the advantages of laparoscopic surgery and has complication rates which compare favourably with other open and laparoscopic techniques. PMID- 10830602 TI - A simplified method of harvesting and dilating the radial artery achieves acceptable clinical outcomes. AB - BACKGROUND: The recent successful revival of the radial artery as a coronary bypass conduit has been attributed to a minimally traumatic harvesting technique without diathermy, combined with long-term oral calcium antagonist therapy. We describe a simplified technique of harvesting the radial artery, which reduces procurement time and maintains conduit relaxation. METHODS: Radial arteries were harvested using diathermy and topical glyceryl trinitrate-verapamil dilator solution. Postoperatively, intravenous glyceryl trinitrate, but no calcium antagonist was used. The clinical results in the first 100 consecutive patients receiving radial artery grafts (RA group), procured using this technique, were compared with a group of 100 patients receiving saphenous vein conduits (SV group) immediately prior to the introduction of the radial artery at our institution. RESULTS: There were no demographic differences between the two groups, other than the SV group being slightly older. There was one intraoperative death in each group. There was no difference in the rate of peri operative myocardial infarction or length of stay in the intensive care unit. At a median follow-up time of 16 months for the RA group, and 25 months for the SV group, the survival rates were 97 and 94%, respectively. All survivors were in the New York Heart Association class I. In the SV group, two postoperative angioplasties were performed. CONCLUSIONS: These early results suggest that this method of procuring the radial artery using diathermy, glyceryl trinitrate and no postoperative calcium antagonists, is rapid, safe and effective. The continued use of this technique is justified, while awaiting the results of long-term angiographic studies. PMID- 10830604 TI - Glossal abscess: an unusual cause of lingual swelling. PMID- 10830605 TI - Airbag injuries: upper limb fractures due to airbag deployment. PMID- 10830606 TI - Traumatic liver necrosis diagnosed by magnetic resonance imaging. PMID- 10830607 TI - Non-operative management of gunshot injury of liver causing bilhaemia. PMID- 10830608 TI - An unusual case of ascites: pitfalls in diagnosis of malignant peritoneal mesothelioma. PMID- 10830609 TI - Giant pseudopolyposis in inflammatory bowel disease. PMID- 10830610 TI - Selected topics related to occupational exposures. AB - The auditory and nonauditory effects of noise can be quite profound, affecting approximately 15 to 20 million Americans. As with most occupational toxins, recognition and careful assessment of noise exposure are the foundation on which preventive measures and treatment are based. Dosimeters can measure noise exposure over specific time periods. Pure tone air conduction audiometric monitoring should be performed on an annual basis in workers at risk for significant noise exposure. Occupational infectious disease involves far more than hepatitis and tuberculosis. Periodic fever, dermatologic manifestations and other symptoms peculiar to a specific disease may be important clues to an occupationally related exposure. Whereas strict attention to hand washing and isolation are cornerstones of prevention, use of protective gear is mandated in certain situations. Zoonotic disease, agriculture exposure, water transmission, and biologic contaminants in buildings can be important but subtle exposures sources. Recognition of these infections often depends on the alertness of the primary care giver. PMID- 10830611 TI - Selected topics related to occupational exposures. Part II. Occupational infectious diseases PMID- 10830612 TI - Selected topics related to occupational exposures. Part III. Occupational dermatology AB - Occupational skin disorders are very common and are a surprisingly frequent cause of lost work time. Failure to suspect an occupational cause can lead to repeated treatment failure and needlessly prolong patient misery and frustration. Primary care providers play a key role in recognizing possible occupational causes and arranging appropriate education, preventive measures, and treatment. Occupational skin conditions often occur in nonindustrial settings in workers such as hairdressers, health care personnel, and food handlers. Irritant contact dermatitis is by far the most prevalent occupational skin condition and is emphasized in this article. Allergic contact dermatitis, infections, skin cancers, and acneform eruptions may also have significant occupational associations. We will present criteria that suggest on occupational exposure, list common offending agents, and review the clinical presentations and relevant pathophysiology. We provide guidance on a directed history and physical examination and suggest when diagnostic testing is most likely to have value. Finally we outline preventive measures such as contact avoidance, barrier creams, and protective gloves and address therapy and indications for referral. PMID- 10830613 TI - Selected topics related to occupational exposures. Part IV. Occupational liver disease AB - Occupational and environmental substances associated with liver injury include industrial chemicals, drugs, certain bacterial and viral infections, and other physical agents. Hepatotoxic chemical agents can be classified as direct hepatotoxins, indirect hepatotoxins, or agents that cause liver injury as a result of host idiosyncrasy. More than 100 industrial chemicals have been shown to be acutely hepatotoxic. Occupations with hepatotoxin exposures are numerous and include farm workers, chemists, dry cleaners, electroplaters, garage workers, health care workers, nurses, painters, printers, rayon makers, and others. Clinical presentation of occupational liver disease may be acute/subacute or chronic but is often insidious. Some hepatotoxins are capable of causing malignancy. The key to diagnosis of occupational liver disease is exposure history. The occupational and environmental history should include a brief description of the patient's current and recent jobs to the extent necessary to assess their potential for workplace hepatotoxic exposures. Confounders such as obesity, alcohol, and viral hepatitis are common, and a careful history and examination are essential. A variety of tests are used to evaluate liver disease and may include serum markers, tests of synthetic liver function, clearance tests, and anatomic tests. A key responsibility of the primary care provider is to prevent further liver injury from preventable occupational exposures; not only the patient but also coworkers may be at risk. Efforts should be made to ensure that patients with potential hepatotoxic exposures are working or living in safe conditions. Collaboration with an occupational specialist may be especially useful in this regard. PMID- 10830614 TI - Selected topics related to occupational exposures. Part V. Occupational cardiovascular disease AB - Cardiovascular disease is common in the United States. Several occupational exposures, such as carbon disulfide and organic nitrates, are believed to cause occupational cardiovascular disease. In addition some other agents, such as lead and cadmium, may indirectly cause cardiovascular disease through their effects on blood pressure. For other agents (ie, carbon monoxide, solvents, and chlorofluorocarbons), acute exposure and high levels may cause cardiovascular disease but may not cause cardiovascular disease through long-term or low levels. A primary care physician who has a patient with a new or unstable cardiovascular disease should obtain an occupational history to assess whether occupational exposures may be playing a role. An occupational history may indicate potential cardiovascular risks. Such risks can include exposure to certain chemicals and metals, physical factors, exertion, or psychological stress. The primary care physician should be able to assess the situation and advise the patient, as well as the employer, about restrictions or accommodations that may need to be made. PMID- 10830615 TI - Expression of MacMARCKS restores cell adhesion to ICAM-1-coated surface. AB - To evaluate the role of MacMARCKS, a major substrate of protein kinase C, in cell adhesion, we selected a macrophage cell line, Wehi 274.1.7. Although surface expression of beta2-integrins can be detected on these cells, they lack the phorbol ester- or chemokine-induced adhesion to ICAM-1-coated surface, an event mediated by beta2-integrins. Concomitantly, these cells lack expression of both MacMARCKS and its homologue, MARCKS. When wild type MacMARCKS was expressed in these cells, the phorbol ester-induced adhesion to ICAM-1-coated surface increased approximately 5-fold compared to vector transfected control cells. To further investigate the potential physiological role of MacMARCKS in this adhesion event, we also tested the effect of monocyte chemotactic protein-1, and a 3-fold increase in the adhesion to ICAM-1-coated surface was observed with MacMARCKS-transfected cells. Therefore, these data suggest that MacMARCKS is an essential component in regulating cell adhesion. PMID- 10830616 TI - Quantitative determination of gap junction intercellular communication by scrape loading and image analysis. AB - Gap junction intercellular communication (GJIC) consists of intercellular exchange of low molecular weight molecules. Chemically induced alterations of this communication have been suggested to result in abnormal cell growth and tumour promotion. Several in vitro assays have been developed to determine the effect of chemicals on gap junction communication in cultured cells. The scrape loading dye transfer technique is based on studying the transfer of the fluorescent dye Lucifer Yellow in cells where the dye is loaded through a cut in the cell monolayer. This technique is rapid and relatively uncomplicated, but has only been used to qualitatively demonstrate communication, due to lack of an appropriate method for quantification of the dye spreading. We show here that analysis of digital fluorescence images of cells scrape loaded with Lucifer Yellow can be used for quantitative determination of GJIC. We have analysed the images both by means of distance of diffusion of the dye in the cell monolayer, as well as by area of dye-coupled cells. The results are consistent with that obtained using microinjection of Lucifer Yellow and the method offers a simple way for quantitative determination of GJIC. PMID- 10830617 TI - Expression of a soluble functional form of the integrin alpha4beta1 in mammalian cells. AB - The integrin alpha4beta1(VLA4) has been expressed as a soluble, active, heterodimeric immunoglobulin fusion protein. cDNAs encoding the extracellular domains of the human alpha4 and beta1 subunits were fused to the genomic DNA encoding the human gamma1 immunoglobulin Fc domain and functional integrin fusion protein was expressed as a secreted, soluble molecule from a range of mammalian cell lines. Specific mutations were introduced into the Fc region of the molecules to promote alpha4beta1 heterodimer formation. The soluble alpha4beta1 Fc fusion protein exhibited divalent cation dependent binding to VCAM-1, which was blocked by the appropriate function blocking antibodies. The apparent Kd for VCAM-1 binding were similar for both the soluble and native forms of alpha4beta1. In addition, the integrin-Fc fusion was shown to stain cells expressing VCAM-1 on their surface by FACs analysis. This approach for expressing soluble alpha4beta1 should be generally applicable to a range of integrins. PMID- 10830618 TI - Tumor-derived mutated E-cadherin influences beta-catenin localization and increases susceptibility to actin cytoskeletal changes induced by pervanadate. AB - E-cadherin participates in homophilic cell-to-cell adhesion and is localized to intercellular junctions of the adherens type. In the present study, we investigated the localization of adherens junction components in cells expressing mutant E-cadherin derivatives which had been previously cloned from diffuse-type gastric carcinoma. The mutations are in frame deletions of exons 8 or 9 and a point mutation in exon 8 and affect the extracellular domain of E-cadherin. Our findings indicate that E-cadherin mutated in exon 8 causes beta-catenin staining at lateral cell-to-cell contact sites and, in addition, abnormally located beta catenin in the perinuclear region. Moreover, the various mutant E-cadherin derivatives increased the steady-state levels of alpha- and beta-catenin and were found in association with these catenins even after induction of tyrosine phosphorylation by pervanadate. Sustained pervanadate treatment led, however, to rounding-up of cells and induction of filopodia, changes which were first detectable in cells expressing E-cadherin mutated in exon 8. The deterioration of the cell contact was not accompanied with disassembly of the E-cadherin-catenin complex. Based on these observations, we propose a model whereby in the presence of mutant E-cadherin tyrosine phoshorylation of components of the cell adhesion complex triggers loss of cell-to-cell contact and actin cytoskeletal changes which are not caused by the disruption of the E-cadherin-catenin complex per se, but instead might be due to phosphorylation of other signaling molecules or activation of proteins involved in the regulation of the actin cytoskeleton. PMID- 10830619 TI - Rac is essential in the transformation of endothelial cells by polyoma middle T. AB - Expression of the Polyoma Middle T (PyMT) antigen in endothelial cells results in single-step transformation to hemangioma producing malignant cells. To study the mechanism of PyMT transformation, we used the PyMT induced mouse brain endothelial cell line, bEND.3, expressing constitutively active and dominant negative mutants of the small GTPase Rac. The bEND.3 cell phenotype of tumorigenesis, loss of normal growth control and formation of cysts rather than capillary tubes in fibrin gels was reversed by expression of dominant negative Rac. The mechanism of N17 Rac action in blocking the endothelial cell transformant, PyMT, did not involve effects of Rac on the actin cytoskeleton since this component of the bEND.3 cell phenotype was not affected. Furthermore, the PyMT induced activation of the plasminogen activator (PA)/plasmin system was not affected by Rac inhibition. Inhibition of the downstream effectors of Rac, phosphatidylinositol 3-kinase (PI3-K) and p70S6k, which are known to be constitutively activated by PyMT transformation, inhibited bEND.3 cell proliferation and cyst formation in fibrin gels even in cells expressing V12 constitutively active Rac, but they did not restore capillary tube formation. These results demonstrate that middle T antigen induced endothelial cell transformation requires signal transduction by Rac. The downstream Rac effectors, P13-K and p70S6k, mediate PyMT/Rac effects on cell proliferation and cyst formation, but other unknown effectors of PyMT are required for the cytoskeletal changes and activation of the PA/plasmin system. PMID- 10830620 TI - Differentiation, proliferation and adhesion of human neuroblastoma cells after treatment with retinoic acid. AB - Because of the known property of spontaneous regression in stage IVS of neuroblastoma all attempts are made to elucidate whether differentiation inducers possibly could be applied for neuroblastoma therapy. Here we examined the influence of retinoic acid (RA) in vitro on differentiation, proliferation and adhesion of 10 permanent and 4 primary cell lines as well as of several SCID mouse tumour transplants. In general, after RA treatment morphologically different cell types which are characteristic for neuroblastoma cells have changed. N (neuronal)-type cells prolonged their neuronal processes, whereas S (epithelial, substrate-adherent, Schwann cell-like)-type cells lost their adherence to substratum and became apoptotic. Additionally, the reactions of all neuroblastoma cell lines with monoclonal antibodies against beta-tubulin (for neuronal cells) and glial fibrillary acidic protein (for epithelial cells) were determined. The anti-proliferative effect of all-trans-RA as well as 13-cis-RA was more profound in S-type cells (up to 40% in primary cell lines). To elucidate the role of adhesion molecules during neuronal cell differentiation, we have analysed the adhesion of neuroblastoma cells on poly-D-lysin-precoated plates under RA influence. While N-type cells displayed an increased adhesion, all S type cell lines as well as all primary cell lines exhibited a reduced adhesion (IMR-5 and IMR-32: p < 0.001; JW, SR and PM: p < 0.05). RA treatment increased predominantly the tested antigens (HCAM, ICAM-1, NCAM, PECAM-1, VCAM-1, cadherin, FGF-R, IGF-R, NGF-R, TGF-beta/1, NF200, NF160, NF68, NSE, HLA-ABC) in all cell lines independently of their phenotypes (TGF-beta/1: p < 0.001; NF68: p < 0.01; PECAM-1 and NGF-R: p < 0.05). In recultured SCID-mouse-passaged tumour cells antigens were down-regulated (FGF-R: p < 0.01), but increased again after RA influence (TGF-beta/1: p < 0.05). In summary, the RA differentiation model demonstrates the possibility to interfere in cell adhesion and to diminish growth potential both in N-type as well as S-type neuroblastoma cells. PMID- 10830621 TI - Popliteal artery entrapment syndrome: diagnosis and management, with report of three cases. AB - Popliteal artery entrapment syndrome is an important albeit infrequent cause of serious disability among young adults and athletes with anomalous anatomic relationships between the popliteal artery and surrounding musculotendinous structures. We report our experience with 3 patients, in whom we used duplex ultrasonography, computed tomography, digital subtraction angiography, and conventional arteriography to diagnose popliteal artery entrapment and to grade the severity of dynamic circulatory insufficiency and arterial damage. We used a posterior surgical approach to give the best view of the anatomic structures compressing the popliteal artery. In 2 patients, in whom compression had not yet damaged the arterial wall, operative decompression of the artery by resection of the aberrant muscle was sufficient. In the 3rd patient, operative reconstruction of an occluded segment with autologous vein graft was necessary, in addition to decompression of the vessel and resection of aberrant muscle. The result in each case was complete recovery, with absence of symptoms and with patency verified by Doppler examination. We conclude that clinicians who encounter young patients with progressive lowerlimb arterial insufficiency should be aware of the possibility of popliteal artery entrapment. Early diagnosis through a combined approach (careful physical examination and history-taking, duplex ultrasonography, computerized tomography, and angiography) is necessary for exact diagnosis. The treatment of choice is the surgical creation of normal anatomy within the popliteal fossa. PMID- 10830622 TI - Gating of myocardial perfusion imaging for the identification of artifacts: is it useful for experienced physicians? AB - Gating of myocardial perfusion imaging helps to differentiate artifacts from perfusion defects. We used this technique to evaluate the impact of routine gating on the interpretation of results by physicians experienced in this field. We studied, prospectively, 270 consecutive patients (161 men and 109 women) who underwent gated myocardial perfusion imaging. Single-photon emission-computed tomography was performed to evaluate myocardial perfusion in patients at rest and after stress, using technetium-99m sestamibi and post-stress gating. Participating physicians interpreted each study and indicated a confidence level for the interpretation. Initially, these opinions were formed on the basis of static slices alone and subsequently, with the addition of gating information. The impact of gating was evaluated by the number of studies in which gating led to a change in interpretation from normal to abnormal or vice versa, or from borderline to definite. The interpretation was changed from abnormal to normal or vice versa in 10 studies (3.7%) and from borderline to definite in 3 (1.1%). In 37 studies (13. 7%), the confidence level was increased from confident to very confident with no change in interpretation. We conclude that routine gating of every myocardial perfusion imaging study for the identification of artifacts is of low value for physicians experienced in interpreting such studies. Although gating frequently increases the confidence level, it seldom leads to a change in interpretation. Specific subgroups of patients who would benefit from gating should be identified. PMID- 10830623 TI - Combined coronary artery and abdominal aortic surgery without cardiopulmonary bypass. AB - To determine the effects of beating heart surgery on patients undergoing simultaneous coronary artery bypass grafting and abdominal aortic surgery, we performed such surgery on 20 patients (mean age, 64.55+/-796 SD years). Abdominal aortic disease was defined as an abdominal aortic aneurysm larger than 5 cm in diameter or as end-stage aortic occlusive disease. Hemodynamic measurements, inotropic requirements, and incidence of perioperative myocardial infarction and arrhythmias were recorded, as were subsystem clinical outcomes, length of intensive care unit and hospital stays, blood loss, and transfusion requirements. There was no incidence of death, perioperative myocardial infarction, stroke, or acute renal failure. The mean number of grafts per patient was 1.95+/-0.69. Only 4 minor postoperative complications were observed: three patients (15%) had evidence of supraventricular tachyarrhythmias, and 1 patient (5%) had chest infection that required a longer-than-average intubation period. Six patients (30%) required minimal-to-moderate inotropic support. The mean blood loss was 673+/-246.8 mL and transfusion requirements were low. The mean intensive care unit and hospital lengths of stay were 2. 12+/-0.33 days and 708+/-1.44 days, respectively. Clinical follow-up (mean, 10 months) showed all patients to be in New York Heart Association functional class I or II with no late cardiac or abdominal events. We conclude that simultaneous coronary artery bypass grafting and abdominal aortic surgery on the beating heart is safe and effective, and has a low perioperative clinical morbidity rate. To our knowledge, ours is the 1st report on this procedure. Larger studies with longer follow-up are needed. PMID- 10830624 TI - Long-term results of apico-aortic valved conduit for severe idiopathic hypertrophic subaortic stenosis. AB - We report our long-term results of apico-aortic conduit implantation in patients with isolated idiopathic hypertrophic subaortic stenosis. Between December 1977 and July 1983, apico-aortic prosthetic-valved conduits were implanted in 4 such patients (age range, 24-65 years) who had severe left ventricular hypertrophy and small left ventricular chambers. In this procedure, the distal end of the conduit was anastomosed to the ascending aorta in 3 patients and to the upper abdominal aorta in 1. Postoperative echocardiography showed relief of the left ventricle aortic gradient and enlargement of the left ventricular chamber in all cases. One patient died of perioperative wound infection. One patient died of unnatural causes 13 years after the initial operation; in his case, the conduit was known to be occluded. Two patients are alive 15 and 19 years, respectively, after the initial operation. Three instances of conduit obstruction due to bioprosthetic calcification were observed. Despite the high incidence of reoperation due to conduit valve failure, apicoaortic conduit implantation has produced good hemodynamic outcome and has improved the quality of life in patients who have idiopathic hypertrophic subaortic stenosis and anatomic features unsuitable for Morrow's operation. Improvements in bioprostheses and in apical implantation techniques may allow a revival of apico-aortic conduit implantation in selected patients with idiopathic hypertrophic subaortic stenosis. PMID- 10830625 TI - Use of selective hypothermia to protect the spinal cord during resection of thoracoabdominal aneurysms. AB - Neurologic complications remain a threat to patients after repair of thoracoabdominal thoracic aneurysms. We have recently used a simple technique of selective hypothermia during such repairs in 7 high-risk patients. None of the patients had neurologic complications. In the 1st patient, we used cardiopulmonary bypass, but we have subsequently found that the procedure can be performed safely without an oxygenator in the circuit by using only a centrifugal pump, heat exchanger, thermistor, and pressure gauge. We believe that this technique deserves additional consideration for avoiding neurologic complications after repair of such lesions. PMID- 10830626 TI - Manubrium-sparing median sternotomy as a uniform approach for cardiac operations. AB - We used a manubrium-sparing sternotomy to perform intracardiac operations on 26 patients between November 1997 and April 1998. We developed this less-invasive surgical technique as a uniform approach in order to reduce skin and skeletal trauma, while maintaining the advantages of the full median sternotomy, such as standard aortic and venous cannulations and use of both antegrade and retrograde cardioplegia. During the same period, 26 other patients with intracardiac lesions underwent operation through a standard full sternotomy. In the manubrium-sparing sternotomy group, there was no intraoperative complication or conversion to full median sternotomy. The average postoperative chest drainage was less in the manubrium-sparing sternotomy group (242.7+/-184.5 mL/24 hours, vs. 499.2+/-416.3 mL/24 hours; P<0.01). Two patients (77%) in the manubrium-sparing sternotomy group had superficial wound disruption, but 4 patients (15.4%) in the full sternotomy group had more severe wound infection, and 1 required myoplasty because of deep wound infection. During the mean follow-up period (12.4+/-1.9 months), no patient in the manubrium-sparing sternotomy group reported significant discomfort or pain due to the sternotomy, but 6 patients (23.1%) in the full sternotomy group complained of significant sternal pain, while 4 (15.4%) experienced shoulder pain, and 1 (3.8%) experienced numbness of the 4th and 5th fingers of both hands. We conclude that the manubrium-sparing sternotomy is a safe and useful approach for most cardiac operations. It is effective in reducing surgical trauma and postoperative wound discomfort. PMID- 10830627 TI - Reconstruction of the superior vena cava with the aid of an extraluminal venovenous jugulo-atrial shunt. AB - A 57-year-old woman had chronic benign superior vena cava syndrome related to the long-term use of multiple central venous catheters for chemotherapy. Treatment included resection of the obstructed segment and repair of the superior vena cava with an autologous pericardial patch. Intraoperatively, return venous flow was maintained with an extraluminal venovenous jugulo-atrial shunt. The shunt relieved upper-body hypertension and congestion, resulting in early extubation and a short, smooth postoperative course. PMID- 10830628 TI - Hemitricuspid dysplasia in association with ventricular septal defect. AB - The tricuspid valve essentially consists of a straight septal leaflet and a curved mural leaflet that includes the anterior and posterior leaflets. A plane bisecting the septal and anterior leaflets divides the chordopapillary support of the tricuspid valve into 2 groups. One group is related to the bulbar and adjacent perimembranous and sinus components of the interventricular septum. The other is related to the trabecular septum and the adjacent free wall. We report our surgical findings in 5 patients who had dysplasia of the leaflets and chordopapillary support related to the bulbar, perimembranous, and sinus portions of the interventricular septum, in conjunction with a perimembranous ventricular septal defect. Phylogenic, ontogenic, and surgical implications of this association are discussed. PMID- 10830629 TI - Coconut atrium: transmural calcification of the entire left atrium. AB - Massive calcification of the left atrium usually spares the interatrial septum, which provides a cleavage plane for surgical access to the mitral valve. Endoatriectomy with mitral valve replacement is the currently accepted corrective procedure because it affords maximum exposure while decreasing the risk of embolization and intraoperative hemorrhage. We describe a case in which the entire left atrium, including the septum, was thickly calcified and resembled a coconut shell. This condition prevented surgical correction of severe mitral stenosis. To our knowledge, this is the most severe case of left atrial calcification yet reported in the literature. Although it is not possible to establish preoperatively that the atrium is completely calcified and impossible to incise, when predisposing factors and evidence of complete transmural calcification are present, the surgeon should be aware of this possibility and should weigh carefully the decision to operate. PMID- 10830630 TI - Cryoablation of incessant ventricular tachycardia: case report and long-term follow-up. AB - We report the case of a 52-year-old man who had incessant ventricular tachycardia despite treatment with antiarrhythmic agents. Placement of an implantable cardioverter/defibrillator, radiofrequency ablation, and antitachycardia pacing were ineffective. He underwent intraoperative arrhythmia mapping and cryoablation at our institution. Eighteen months later, he remained free of arrhythmia. He was in New York Heart Association functional class II with an ejection fraction of 30%. PMID- 10830631 TI - Minimally invasive parasternal approach to tricuspid valve avoids repeat sternotomy. AB - We report the case of a 68-year-old patient with severe tricuspid regurgitation who had previously undergone aortic valve replacement and right coronary artery bypass. We performed tricuspid valvuloplasty via the right parasternal route in order to reduce surgical trauma by avoiding resternotomy, trauma to the venous graft, and bleeding due to dissection of old adhesions. The patient's postoperative course was uneventful, and he was discharged home on the 7th postoperative day. PMID- 10830632 TI - Lymphocutaneous fistula as a long-term complication of multiple central venous catheter placement. AB - We report a case of a lymphocutaneous fistula in a 19-month-old boy who had been a premature neonate, born in the 23rd week of gestation. The fistula, an apparent complication of central venous line placement during the patient's first 5 months of life, was composed of a distinct lymphatic vessel bundle in the right supraclavicular region, with its exit point at the posterior aspect of the right shoulder. The drainage ceased immediately after resection and repair of a 1-cm obstruction in the superior vena cava. PMID- 10830633 TI - Surgical treatment of right atrial myxoma. AB - A 51-year-old man with a large right atrial myxoma underwent emergency surgical resection in our institute. The diagnosis of such tumors can be difficult, and their resection presents difficulties for the placement of the venous cannulae. We used a single cannula in the superior vena cava until fibrillation, and then we inserted a cannula into the inferior vena cava. We present this technique as a method of avoiding embolization. PMID- 10830634 TI - Aorto-right artrial fistula: a rare complication of aortic dissection. AB - We describe the successful surgical repair of an acute aortic dissection that had caused an aorto-right atrial fistula in a 67-year-old man. The patient was admitted to the hospital on an emergency basis because of severe heart failure. The diagnosis of acute aortic dissection with rupture into the right atrium was confirmed by use of intraoperative transesophageal echocardiography, although rupture of a sinus of Valsalva aneurysm into the right atrium had been suggested initially by 2-dimensional and Doppler transthoracic echocardiography. At surgery, we found the patient to have aortic arch dissection with complete separation of the right coronary artery from the sinus of Valsalva and a false lumen that had ruptured into the right atrium. The aortic arch was repaired directly. The ascending aorta was successfully replaced with a composite graft. Aortic dissection with rupture into the right atrium is extremely rare and leads to death rapidly. As shown in this case, such a condition might be mistaken for an aneurysmal rupture of the sinus of Valsalva, with use of transthoracic echocardiography alone. Transesophageal echocardiography is a useful noninvasive method to further define or confirm the diagnosis. Early surgical intervention is necessary in patients with this condition to prevent profound shock and end-organ failure. PMID- 10830635 TI - Mitral and aortic valve endocarditis due to Staphylococcus lugdunensis. AB - Staphylococcus lugdunensis is a recently described coagulase negative staphylococcal species involved in human infections. Endocarditis caused by Staphylococcus lugdunensis has been reported rarely: fewer than 50 cases have been described so far. The infection is frequently complicated by embolic events and carries a high mortality rate. We report a case of endocarditis due to Staphylococcus lugdunensis in which the native mitral and aortic valves were infected. The bacterium was isolated on cultures from the aortic and mitral vegetations. Appropriate medical and surgical treatment led to a good outcome of the infection. At 6-year follow-up, there was no evidence of recurrence, and the patient showed good functional recovery. He was in New York Heart Association functional class I. PMID- 10830636 TI - Proximal internal mammary in situ graft and distal coronary-coronary graft. AB - When a left anterior descending coronary artery passes over the cardiac apex and presents with 2 stenoses, 1 proximal and 1 distal, the available bypass conduit often is too short to enable both the anastomosis below the distal stenosis and the sequential anastomosis on the arterial segment between the 2 stenoses. In this circumstance, we graft the internal mammary artery in situ onto the proximal segment of the left anterior descending coronary artery, then use a short residual segment of the internal mammary to perform a coronary-coronary bypass of the distal stenosis. This technique also spares segments of the internal mammary for other purposes. We present our experience, together with angiographic evidence of long-term patency. PMID- 10830637 TI - Atherosclerotic coronary artery aneurysms: eight-year angiographic follow-up. PMID- 10830638 TI - Rapid lysis of deep vein thrombosis by low molecular weight heparin. PMID- 10830639 TI - Transmyocardial revascularization: could mechanical puncture be more effective than puncture by laser? PMID- 10830640 TI - The significance of a patent foramen ovale. PMID- 10830641 TI - Six-year survival of unoperated ventricular septal rupture following myocardial infarction. PMID- 10830642 TI - Deprivation and oral health: a review. AB - The link between socioeconomic status and health, including oral health, is well established. The conventional measures of socioeconomic status used in these studies, such as social class and household income, have a number of weaknesses so that alternatives, in the form of area-based measures of deprivation, are increasingly being used. This paper reviews epidemiological research linking deprivation and oral health. Four types of study are identified and described: simple descriptive, comparative, analytic and explanatory. These studies confirm that deprivation indices are sensitive to variations in oral health and oral health behaviours and can be used to identify small areas with high levels of need for dental treatment and oral health promotion services. As such, they are likely to provide a useful administrative tool. In terms of research, the studies demonstrate that these measures provide a ready way of controlling for socioeconomic status in studies examining the association between oral health and other variables. However, this research, in largely replicating previous studies using social class, does not address fundamental issues concerning the mechanisms which link social inequality and health. Deprivation measures have a major role to play in research that examines features of people and places, and how they promote and/or damage both oral and general health. PMID- 10830643 TI - Efficiency in provision of public dental services in Norway. AB - The paper examines productivity in the provision of public dental services in Norway. The main responsibility of the public dental services is to provide care for all children up to 18 years of age. Most dental care is provided by public dental officers. Productivity was measured by estimating a stochastic production frontier from input and output data from the public dental service in Norway. The results indicate that there are increasing returns to scale. Our measure of technical efficiency showed that the level of inefficiency is fairly small. However, the inefficiency is greater when estimated from a deterministic production frontier rather than from a stochastic frontier. One limitation of the present methodology is that it does not say anything about the level of efficiency in the Norwegian public dental service in absolute terms. A greater level of inefficiency would have been identified if a few counties had performed clearly better than the rest. PMID- 10830644 TI - Comparison of three salivary flow rate assessment methods in an elderly population. AB - Measuring salivary flow rates among the frail elderly is a challenge. The currently used spit collection method requires levels of time and cooperation that often may exclude the frail elderly who are at high risk for salivary compromise. A measurement method that is not only valid and reliable, but also feasible and acceptable is needed for use in population studies of compromised adults. This study compared two salivary flow rate assessment methods using a suction machine against the currently accepted spit collection method in an elderly population aged 75 and older. Three methods of flow rate (g/min) assessment were compared at three time periods among 16 elders (mean age 86.6 years). Flow rates using the 2-min open suction method compared well with the 10 min spit method (r=0.778) but the 2-min closed suction method did not (r=0.158). Reliability evaluation of the open suction method and the spit method was assessed using a test/retest with a 1-week interval. Both methods demonstrated good comparable reliability (spit method r=0.566, P=0.01); open suction method, (r= 0.861, P<0.01). Based on a short survey questionnaire about the three methods, 11 of the 16 elderly subjects preferred the use of the suction machine to the spit method. These results indicate that the 2-min open suction method technique is a valid and reliable means of measuring salivary flow. The lower level of patient cooperation needed, the shorter time period required, and this preliminary report of its acceptability support the use of this method in future population studies of frail elders. PMID- 10830645 TI - Dentist-related factors influencing the amount of prosthodontic treatment provided. AB - OBJECTIVES: To evaluate, using multivariate methods, the associations between indicators of the amount of prosthodontic treatment and dentist-related factors. METHODS: Questionnaires were sent to a random sample of 2,059 general dentists, response rate was 76%. Two indicators of prosthodontic activity were used as dependent variables: 1) reported weekly working hours used for prosthodontics and 2) reported numbers of produced single crowns, fixed partial dentures, and removable dentures. Independent variables were 'social and demographic attributes', 'job situation' and 'attitudes of dentists'. Multiple regression analysis was used in models with continuous dependent variables and logistic regression analysis for categorical dependent variables. RESULTS: 'Weekly working hours used for dental care of adults' showed a strong association in all models with the dependent variable 'weekly working hours used for prosthodontics'. Male dentists provided more prosthodontic services than female dentists, even if reporting less time used for prosthodontics. Private practitioners produced more fixed prosthodontics than dentists employed in the public dental health service. Dentists in the public dental health service reported a higher production of removable dentures than private practitioners. CONCLUSION: The results indicate that factors, besides those in the rational clinical model for decision-making, e.g. gender and delivery system, play a role in the provision of prosthodontic services. PMID- 10830646 TI - Development of a condition-specific quality of life measure for patients with dentofacial deformity: I. Reliability of the instrument. AB - The assessment of quality of life is becoming increasingly important in clinical research. Its importance in dentistry has been realised only relatively recently. Health-related quality of life is concerned with the aspects of quality of life that relate specifically to an individual's health. This may be measured using two groups of instruments: (i) generic measures, which provide a summary of health-related quality of life and sometimes generate a single index measure of health or (ii) condition-specific measures, which focus on a particular condition, disease, population or problem and are potentially more responsive to small, but clinically important, changes in health. OBJECTIVES: The aim of this study was to develop a condition-specific quality of life measure for those patients with severe dentofacial deformity who were requesting orthognathic treatment and to assess the reliability of this instrument. METHOD: Instrument content was derived through a literature review and interviews with clinicians and patients. The resulting instrument was tested for internal consistency and test-retest reliability. RESULTS AND CONCLUSION: The instrument was found to divide into four clinically meaningful domains. Internal consistency and test retest reliability were good. Patient acceptance of the questionnaire was also encouraging. PMID- 10830647 TI - Impact of analytic conventions on outcome measures in two longitudinal studies of dental caries. AB - Caries increment is the primary outcome measure used in most longitudinal studies of caries. Its computation requires the adoption of specific analytic conventions for handling examiner misclassification ("reversals"), teeth lost due to caries, and multiple events such as caries initiation and progression. However, little is known about the impact of these analytic conventions on cumulative incidence and incidence density--two additional outcome measures used in longitudinal studies. OBJECTIVE: We evaluated the impact of analytic conventions on caries outcome measures in two longitudinal caries studies. METHODS: A secondary analysis was undertaken using data from two studies: a 2-year observational cohort study of 683 older adults and a 3-year clinical trial of 1,754 children. Crude, net and adjusted increments were compared, effects of inclusion and exclusion of teeth lost due to caries were contrasted and measures of caries initiation and progression were assessed among subgroups in each study. RESULTS: Cumulative incidence indicated a high risk of caries (50% or more) in both studies but was not sensitive to subgroup differences. Increment and incidence density measures consistently revealed between-subgroup differences when subgroups had comparable numbers of teeth. For all three outcome measures, crude DFS values were larger than net DFS values. The adjusted DFS was similar to crude DFS in older adults, but similar to net DFS in children. Measurement of caries initiation and progression increased disease estimates for all three outcome measures. CONCLUSIONS: In the populations studied, analytic conventions for handling examiner misclassification, teeth lost due to caries and multiple events had a greater impact on inferences than the choice of outcome measure. PMID- 10830648 TI - A clinical index for evaluating and monitoring dental erosion. AB - This study describes a new fine-scaled system for classifying initial and advanced dental erosions. The system includes the use of study casts of the teeth in an epoxy resin with an accurate surface reproduction. The severity of erosion on each tooth surface is scored according to six grades of severity. In addition, the presence of a Class V restoration and dental erosion on the same surface increases the erosion score, as it is assumed that the need for restorative treatment can be caused by the erosion. A high inter-examiner agreement was found when the present scoring system was used by two examiners on the same sample. With this prerequisite it is proposed that an index value for facial, oral, incisal/occlusal and cervical surfaces is calculated as the mean value of scores for the respective surfaces. The index values represent the severity of tooth substance loss in various locations of the oral cavity and are furthermore suitable for data analysis. The system is thereby well-suited for determining etiologic factors and monitoring the progression of erosion over time. PMID- 10830649 TI - Xylitol candies in caries prevention: results of a field study in Estonian children. AB - All field studies have unequivocally reported significant reductions in dental caries occurrence associated with the use of chewing gum containing xylitol. No other xylitol products besides chewing gum have so far been tested in field trials. A 5-year follow-up study with 2- or 3-year xylitol consumption periods began in Estonia in 1994 with 740 10-year-old children in 12 schools at baseline examinations. For the study, 3 clusters each including 3-5 schools were formed on the basis of baseline caries experience. The products were used under the supervision of the teachers 3 times per day during school days but not during weekends or during the 3-month summer holiday. The daily dose of xylitol was 5 g in all groups. The children were examined every year in September by two experienced clinicians. Dental caries was recorded according to WHO criteria. After 3 years, all xylitol groups showed a highly significant 35%-60% reduction in caries incident, compared with the corresponding control groups. The differences between candies, between candies and chewing gum, and between 2- and 3-year users in the xylitol groups were non-systematic, indicating no trends between the groups. The results suggest that not only xylitol chewing gum but also xylitol candies are effective in caries prevention, and that a school-based delivery system seems to offer a practical way to distribute and control the use of the xylitol products. PMID- 10830650 TI - Service provision patterns by main diagnoses and characteristics of patients. AB - Service provision patterns may be influenced not only by clinical oral health status leading to a diagnosis and treatment plan, but also by other variables such as patient characteristics. The main aim of this study was to investigate whether associations between services provided and patient factors would persist after controlling for the main presenting diagnosis or condition. A random sample of dentists surveyed in 1993-94 provided a response rate of 74%. Private general practitioners recorded service provision data from logs of 1-2 typical days of practice. Caries (26.5%) was the most prevalent diagnosis, followed by recall/maintenance care (19.0%), pulpal/periapical infection (10.9%), and failed restorations (10.4%). Diagnoses were associated with variation in the percentage of patients receiving services in main areas of service, and also with insurance status, sex and age distributions of patients, and type of visit (chi-square; P< 0.05). Logistic regressions of receipt of services indicated statistically significant associations with patient characteristics and diagnosis categories. Controlling for diagnosis, uninsured patients and those visiting for emergencies had less favourable service patterns (e.g., higher odds of extractions, but lower odds of preventive and crown and bridge services) compared to patients who had dental insurance or visited for check-ups or other non-emergency dental problems. The influence of these factors on services provided has implications of public health importance in terms of appropriateness of care and social inequality. PMID- 10830651 TI - Trends in caries and associated variables among young Israeli adults over 5 decades. AB - This study was conducted among a representative sample of 7,139 21-year-old Israeli adults, upon release from compulsory military service. Data were collected between 1994 and 1997. The average DMFT level found was 8.49+/-4.95. Untreated caries (according to the D component of DMFT) was 2.25+/-2.90 and significantly higher among males. Untreated caries was also significantly associated with geographic origin: higher among subjects of African or former USSR descent; and with family size: higher among subjects with four or more siblings; with education: caries was higher among subjects with less than 12 years of schooling; and with smoking: caries was more extensive among those who smoked (P<0.0001 for all the associations). Caries severity (DMFT) was found to be significantly associated with father's country of origin and number of siblings (highest among subjects of Asian and African descent and those who had four or more siblings). This is the largest and most representative Israeli survey of young adults to date. Data were compared with available data from studies conducted in 1956, 1966, 1973, and 1985. DMFT scores had previously increased from the 1950s until the 1980s. Treatment levels had increased since the 1980s. The present data, together with results of studies conducted among Israeli children, indicate a recent national decrease in caries experience. This study indicates a further increase in caries treatment level, but a persisting inequity according to social variables. PMID- 10830652 TI - Medical mendacities. PMID- 10830653 TI - The Weil osteotomy of the lesser metatarsals: a clinical and pedobarographic follow-up study. AB - The clinical results with pedobarographic analysis were assessed in 32 patients (59 metatarsals) who underwent a distal metatarsal shortening (Weil) osteotomy for either intractable plantar keratoses or chronically dislocated lesser metatarsal phalangeal joints. All patients had increased pressure under the involved metatarsal heads. Thirty three of the 59 metatarsophalangeal (MTP) joints were chronically dislocated. At an average follow-up of 30 months, patients rated the result as excellent or good for 32 of the 37 feet (86%). The mean preoperative AOFAS score was 59 (maximum 100), which improved to 81 post operatively. This difference is significant: p = 0.00001 (with t-test). Comparison of the pre and post-operative pedobarographic measurements showed a significant decreased load under the affected metatarsal heads (p = 0.05). A complete disappearance of the callus was noted under 44 operated metatarsals (75%) and partial disappearance under 12 metatarsals (20%). Two symptomatic transfer lesions occurred under an adjacent metatarsal head. Recurrent dislocations occurred in 5 joints (15%). While metatarsophalangeal joint range of motion was significantly diminished, toe strength was maintained. Average metatarsal shortening was 5.9 mm with no nonunions, delayed unions, or malunions. The Weil shortening osteotomy is a simple and reliable procedure which can effectively reduce the load under the lesser metatarsophalangeal joints and is helpful for the reduction of dorsally dislocated MTP joints. PMID- 10830654 TI - The medial crosssover toe: a cadaveric dissection. AB - The development of a medial crossover second toe (second toe crossing over the first toe) is not a rare clinical condition. It often occurs in the setting of hallux valgus, although not exclusively so. The resulting displacement of the second toe can cause pain in shoes, with surgical correction being problematic. The pathologic anatomy of this condition has not been fully described. In an effort to better understand it, dissection of a cadaveric specimen with a full crossover toe is presented. The dissection revealed findings not previously documented. They include medial displacement of the flexor tendons and plantar plate along with deformity of the plate itself. These changes are in addition to contracture of the medial collateral ligaments and the previously described rupture of the lateral collateral ligaments. Both the plantar plate and the collateral ligaments, the two major static soft tissue stabilizers of the lesser MP joint, were found to be significantly involved. Pull on the flexor tendons only accentuated the malalignment of the toe. CLINICAL SIGNIFICANCE: The extensive soft tissue changes explain the difficulty in achieving a successful long term correction of a full medial crossover toe with a soft tissue procedure. With attenuation of the plantar plate and medial displacement of the flexor tendons, there is an imbalance of muscle forces across the MP joint. This muscle imbalance would not be corrected by release of the medial collateral ligament, dorsal capsular release or extensor tendon lengthening. Reconstruction of the collateral ligament is at risk for incomplete correction since it is unlikely to resolve deformity in the plate if already present. PMID- 10830655 TI - The injury of the calcaneocuboid ligaments. AB - The selective rupture of the calcaneocuboid ligament is extremely rare and frequently misdiagnosed. This study tries to clarify the mechanism, classification and treatment of this entity. The necessity of radiographs with varus stress and in certain cases of computer tomography (CT) and magnetic resonance imaging (MRI), beside the routine antero-posterior and lateral views, is emphasized. Thirteen cases out of five-hundred-twenty-one sprain injuries of the ankle are described, classified and the therapy discussed: If on varus stress radiographs, there is a calcaneocuboid angle <10 degrees without a bony flake (type 1) strapping for six weeks is indicated. A calcaneocuboid angle >10 degrees with or without a small bony flake of the ligament insertion (type 2) should primarily be treated with a shoe cast for 6 weeks; if there are persistent symptoms a secondary peroneus brevis tendon graft is recommended. A calcaneocuboid angle >10 degrees with a big flake (type 3) should be treated by open reduction and refixation of the ligament. Complex injuries (type 4) are characterised by cuboid compression fracture and ligament rupture. PMID- 10830656 TI - Medial impingement syndrome of the anterior tibiotalar fascicle of the deltoid ligament on the talus. AB - Eleven patients with medial impingement syndrome of the anterior tibio-talar fascicle (ATTF) of the deltoid ligament on the talus were identified. Six sustained inversion injuries, one talus and four ankle fractures. The mean preoperative Ankle-Hindfoot Scale (AHS) was 56 and the postoperative was 87. Surgical debridement of the ATTF was performed. A thickened ATTF and localized synovitis were seen. Talar osteophytes were removed in 8 patients. Mean follow up 4 years. Nine patients had good to excellent results and all patients returned to work. PMID- 10830657 TI - Subtalar arthrodesis for complications of intra-articular calcaneal fractures. AB - Eighty six subtalar arthrodeses performed between 1985 and 1996 for complications associated with intra-articular calcaneal fractures were retrospectively evaluated. Patients were divided into three Groups: (I) 59 patients with calcaneal malunions (II) 13 patients with failed open reduction and internal fixation, and (III) eight patients undergoing open reductions and primary fusion for highly comminuted fractures. In each scenario, internal fixation was achieved with cancellous lag screws. Bone graft material consisted of either autogenous iliac crest graft, local graft obtained from the lateral wall exostectomy of the malunion, or freeze-dried cancellous allograft. Fusions in Groups II and III were performed in situ. Fusions in Group I were performed either in situ or utilizing a variety of reconstructive procedures depending upon the type of malunion encountered. Eighty three of the 86 fusion attempts were successful following the initial operations for a union rate of 96%. Fusion rates were similar regardless of the graft material used. Complications included four varus malunions, four cases of osteomyelitis, and two cases of reflex sympathetic dystrophy. A statistically significant shorter hospital stay was found for patients not undergoing iliac crest bone graft procedures. Eighty patients with at least two year follow up achieved a mean American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot score of 75.0. Scores were similar for all three groups and for the various types of reconstructive procedures used. No correlation was found between postoperative talar declination angle and the AOFAS ankle-hindfoot score. Worker's compensation patients tended to have a poorer clinical outcome. PMID- 10830658 TI - The sustentaculum tali: anatomic, radiographic, and surgical considerations. AB - Three purposes of this study were to: 1) describe the sustentaculum tali in relation to surrounding structures, 2) identify its radiographic projection onto the lateral calca neus, and 3) determine optimal angles for screw placement. Twelve cadaver specimens were used. The length a width of the sustentaculum was measured. Its midpoint was measured from surrounding anatomic landmarks. A point on the lateral calcaneal wall "point A" was chosen. This marks the intersection of two lines bisecting the midpoint of the posterior facet and the lateral calcaneal height. The angles required to pass a pin from point A into the sustentaculum were determined. The sustentaculum projected as a triangular/elliptical structure on the lateral radiograph. It was superimposed on the anterior portion of the posterior facet. PMID- 10830659 TI - Bizarre parosteal osteochondromatous proliferation of bone (BPOP): an unusual foot mass in a child. AB - This report describes an unusual case of bizarre parosteal osteochondromatous proliferation of bone (BPOP) which developed in the foot of an eight year-old child. Also described as Nora's lesion, BPOP is a rare benign bone tumor found most commonly in the hands and feet with a radiographic appearance occasionally confused with malignancy. Resembling osteochondromas at first glance, these lesions have a distinct radiographic and histologic appearance that permits differentiation from other benign lesions or low-grade malignant tumors. Treatment of BPOP consists of simple excision, although there is high rate of recurrence. Despite this high recurrence rate, there have been no reported metastases and local excision is still recommended. PMID- 10830660 TI - Bizarre parosteal osteochondromatous proliferation (Nora's lesion) of the sesamoid: a case report. AB - We report the a case of Nora's lesion (Bizarre Parosteal Osteochondromatous Proliferation) of the sesamoid. A 32-year-old woman presented with a painless, enlarging mass of two years duration on the plantar aspect of the first metatarsophalangeal joint of the left foot. Radiographs, Computerized Tomographs and Magnetic Resonance images, initially suggested a parosteal osteosarcoma arising from the tibial sesamoid. The mass was excised, and a histological diagnosis of Bizarre Parosteal Osteochondromatous Proliferation of bone (Nora's lesion) was made. The aggressive growth of this lesion may suggest a neoplasm clinically. Histological features, however, are those of a reactive lesion. PMID- 10830661 TI - Reconstruction of the lateral ligaments of the ankle using semi-tendinosis graft. AB - Injury to the lateral ligaments of the ankle is one of the most common sports related injuries. Although these injures generally resolve with non-operative treatment reconstruction of the lateral ligaments of the ankle is occasionally necessary. We report the surgical technique and result of reconstruction of the anterior talo-fibular ligament using a semi-tendinosis free graft in 26 patients with an average follow-up of 24 months. 81% of patients reported complete resolution or substantial improvement in instability, swelling and sharp pain. Episodes of functional instability persisted in five patients post operatively. No significant differences were noted between the operated and contra-lateral non operated ankle in range of motion or uniaxial balance assessment. Kin-Com testing of hamstring strength showed no differences between sides. Short term results suggest that this ligament reconstruction provides good results without sacrifice of peroneal function and has minimal donor site morbidity. PMID- 10830662 TI - The effect of axial load on the in vivo anterior drawer test of the ankle joint complex. AB - The anterior drawer test is commonly used in the diagnosis of ankle joint mechanical instability. However, the effect of axial load on the anterior drawer test has not been examined in vivo. The purpose of the study was to assess the effect of axial load on passive anterior instability, and on the diagnostic measurement of the anterior drawer instability of the ankle joint complex. A total of 21 subjects with various degrees of ankle sprains were tested on a device that could continuously record applied anterior force and the resultant displacement of the rear-foot. Anterior drawer flexibility of the ankle joint complex in a neutral dorsi/plantar flexion position was quantified on both feet for all subjects without and with an axial load (385 N). Flexibility of the ankle joint complex in anterior drawer was defined as the slope of a linear load displacement curve (which fitted test data with high correlation coefficients (r>0.991)). With axial load, anterior drawer flexibility was significantly reduced by 28.8% compared to that without axial load. The difference in anterior drawer flexibility between injured and intact ankles significantly decreased with axial load. An axial load increased the stability of ankle joint complex. However, axial load reduced the sensitivity of anterior drawer test to mechanical instability of the ankle joint complex. PMID- 10830663 TI - Retrograde drilling of osteochondral lesions in the mediotalar dome. PMID- 10830664 TI - The National Quality Forum enters the game. PMID- 10830665 TI - Quality and continuity of care in Dutch nurse clinics for people with rheumatic diseases. AB - OBJECTIVE: Recently a new form of nurse clinic for people with rheumatic diseases has been introduced into Dutch health care. This study gives insight into: (i) patients' perceptions about the quality and continuity of care given at these (transmural) nurse clinics; and (ii) specialized rheumatology nurses' and rheumatologists' perceptions about the quality and continuity of care in the clinics. DESIGN: Validated measurement tools (QUOTE and QCC) were used, before and after patients visited a clinic, to determine patient perceptions about the quality and continuity of care. Semi-structured interviews with professionals were used to gather information about their perceptions. SETTING: The study was carried out at five locations in The Netherlands where a home care organization and a general hospital collaborated closely and had joint responsibility for a transmural rheumatology nurse clinic. STUDY PARTICIPANTS: A total of 128 patients, six specialist rheumatology nurses and four rheumatologists. INTERVENTION: Transmural nurse clinics for people with rheumatic diseases. RESULTS: In general, patients were positive about the quality and continuity of care given at the clinics. Some continuity aspects, like the presence of a locum nurse and providing the locum with sufficient information could be improved. Professionals were positive about the information given at the clinics, which is additional to the information given by a rheumatologist. Professionals were less positive about some of the clinics' preconditions. CONCLUSION: In this study, a control group (e.g. patients who received standard rheumatologist care) was not available. However, in comparison with patients' experiences of standard medical care in other (comparable) research, patients' experiences in this study were very positive. It was concluded that Dutch transmural nurse clinics, to a large extent, meet patients' and professionals' expectations and were a positive development in the care of rheumatic patients. PMID- 10830666 TI - Eye examinations for VA patients with diabetes: standardizing performance measures. AB - OBJECTIVE: To demonstrate the potential of the Health Plan Employer Data and Information Set (HEDIS) for the calculation of a performance measure for eye exams in the diabetic population using Veterans Health Administration (VA) administrative data. DESIGN: We calculated a 1-year HEDIS-defined patient denominator and three alternative denominators that considered coding factors in identifying a VA patient as diabetic. We calculated the HEDIS-defined numerator, along with alternative specifications that captured other types of eye exams. Finally, we supplemented national data with VA pharmacy and Medicare claims data to identify all VA diabetic patients at 14 selected VA facilities and to establish a more accurate picture of non-VA health care utilization. RESULTS: The national average annual HEDIS-defined eye exam rate in the VA was 26% in fiscal 1997 compared with 39% for managed care organizations. Medicare utilization raised this by 15 percentage points at 14 northeastern VA hospitals. Over 2 years, at least two-thirds of diabetic VA patients had some type of eye exam through VA or Medicare. CONCLUSION: A HEDIS measure of eye exams for VA patients with diabetes can be calculated using VA administrative data only. However, the question remains to what extent the denominator and numerator accurately and completely identify all diabetic patients using VA services and all appropriate eye exams. We recommend caution in interpreting the results of performance measurement across different health care sectors based on what we currently know are data system limitations. PMID- 10830667 TI - Imbalance of demand and supply for regionalized injury services: a case study in Greece. AB - OBJECTIVE(S): To study regionalized acute injury services on an island with high seasonal fluctuation of the population at risk. STUDY PARTICIPANTS: Of 9432 individuals with traumatic injuries who contacted the hospital during 1996, 1204 were hospitalized. Information was recorded on several injury-related clinical and sociodemographic variables. Possible residual disabilities, 6 months after the injury, were also assessed. MAIN OUTCOME MEASURES: Injury Severity Score (ISS), clinical outcome and duration of hospitalization, odds of transfer to other institutions. RESULTS: Non-residents, whether Greek or foreign nationals are hospitalized for shorter periods. Motor vehicle accident victims are hospitalized on average for 15% longer. Injury victims admitted on a Friday are hospitalized for a longer period. Finally, ISS is a powerful positive predictor of duration of hospitalization. Male injury victims, those injured during late night or early morning and patients injured in July are more likely to be transferred to another institution. Age of the patient and ISS are powerful and independent predictors of an unfavourable outcome. CONCLUSION: The extra demand created by injured tourists is reflected in the seasonality of admissions for injuries. The district hospital of Kerkyra cannot be considered as deficient in comparison to other district hospitals. Nevertheless, the suboptimal function of the hospital, with respect to injuries, is reflected in the high proportion of injured patients transferred when the injury occurs outside the full working schedule of the hospital. Patients with burns, bone fractures or dislocations and head injuries or concussion are transferred with an overall frequency of about 15%--too high to be compatible with a well functioning secondary care institution. PMID- 10830668 TI - Obstacles to collaborative quality improvement: the case of ambulatory general medical care. AB - OBJECTIVE: To assess the effectiveness of inter-site collaboration and report card style feedback of quality measures on quality improvement in the outpatient setting and to identify major barriers to improvement. DESIGN: A collaborative quality improvement effort consisting of a large cross-sectional data collection effort (chart reviews and patient surveys), feedback of comparative quality of care data to improvement teams, and collaboration between sites. SETTING: Eleven primary care sites in the Boston area. STUDY PARTICIPANTS: Quality improvement teams at each site with physician leaders. INTERVENTION: Education about techniques of rapid-cycle quality improvement, coaching of on-site teams, and report-card style feedback of comparative site-specific quality of care data. RESULTS: Multiple quality improvement projects were undertaken through this collaboration. However, though we were careful to educate teams on methods of continuous quality improvement and to name specific clinical leaders, the degree of collaboration and quality improvement fell short of expectations. Major impediments to improvement included lack of team members' time and resources, lack of incentives, and unempowered team leadership. The primary obstacle to collaboration was the diversity of sites and inability of teams to create interventions that were relevant to other sites. CONCLUSION: Despite ample quality of care data, quality improvement education, and a structured collaborative process, achieving quality improvement in the ambulatory setting is still a difficult challenge. Organizations need to find ways of overcoming the obstacles faced by improvement teams in order to maximize quality improvement. PMID- 10830669 TI - Can improving quality decrease hospital costs? AB - PURPOSE: To determine whether the concept of 'cost of quality' and the techniques used for its study in the industrial sector are also applicable to hospitals. DATA SOURCES: We undertook a systematic review of the literature published since 1992 (five electronic databases and a manual search) using keywords relating to quality of health care and costs. STUDY SELECTION: We selected all articles relating cost and quality, providing indicators for quality failure, determining the cost of failure, and itemizing the cost of quality. Twelve articles met these criteria (USA, nine; UK, one; Australia, one; France, one); six referred to total quality management, three to hidden costs, and three to adverse events. DATA ABSTRACTION: For each article, we recorded the test hypothesis, the focus of the study and the main results on costs and quality. RESULTS OF DATA SYNTHESIS: Preventing failure by applying total quality management to a variety of projects (managed care project, setting-up a standard procedure . . .) led to financial savings; quality was maintained, even enhanced. Better communication and co ordination reduced hidden costs and also increased quality. Adverse events prolonged hospital stays by 1.74-4 days and increased costs. CONCLUSION: Very few detailed articles related cost and quality and, although they all noted a positive impact of an emphasis on quality, they nevertheless had their shortcomings. Study periods were too short, the indirect aspects of costs and savings were not taken into account, economic reference values were omitted. We conclude that more precise and strict methods for quantifying costs are needed. PMID- 10830670 TI - Effects of financial incentives on medical practice: results from a systematic review of the literature and methodological issues. AB - OBJECTIVE: To identify all financial incentives that had been proposed, described, or used regardless of their initial objective and, when possible, to assess the results of these incentives on costs, process or outcomes of care. MATERIAL AND METHODS: Systematic review of the literature. Databases searched were: Medline, Embase, Health Planning and Administration, Pascal, International Pharmaceutical Abstracts and the Cochrane Library. Search terms were: health professionals and tape of practice, type of incentive, methodology, languages English or French, January 1993 to May 1999. RESULTS: Financial incentives concerned the modalities of physician payment and financing of the health care system. Confounding factors included: age of the doctor, training, speciality, place and type of medical practice, previous sanctions for over-prescribing, type and severity of disease, type of insurance. Risks of financial incentives were: limited access to certain types of care, lack of continuity of care, conflict of interests between the physician and the patient. Any form of fund-holding or capitation decreased the total volume of prescriptions by 0-24%, and hospital days by up to 80% compared with fee-for-service. Annual cap on doctors' incomes resulted in referrals to colleagues when target income is reached. DISCUSSION: Financial incentives can be used to reduce the use of health care resources, improve compliance with practice guidelines or achieve a general health target. It may be effective to use incentives in combination depending on the target set for a given health care programme. PMID- 10830671 TI - An evaluation of patient satisfaction with family practice care in Slovenia. AB - OBJECTIVE: To describe the level of patient satisfaction with family practice in Slovenia. DESIGN: An internationally developed instrument for patients' evaluations of general practice care was used in a postal survey. SETTING: A representative sample of 36 family practices in Slovenia. STUDY PARTICIPANTS: Sixty consecutive patients in every practice were approached and offered a self administered questionnaire. A total of 2160 questionnaires were handed out. MAIN OUTCOME MEASURE: Percentages of patients reporting level of satisfaction on a 5 point Likert scale for the items in the questionnaire. RESULTS: On average 58.2% of respondents rated the level of care received as excellent. Waiting in the waiting room was the item rated poorest (26.0%). Participants were also less satisfied with perceived time during the consultation (51.6%) and with connectional aspects of care: the feeling that family practitioners showed interest in their personal situation (46.5%); the feeling that family practitioners made it easy to explain problems (49.1%). On the other hand patients praised many other aspects of family practice care in Slovenia: confidentiality of medical records (77.0%); listening capacity of their family physicians (69.4%); being able to speak to the family practitioner on the 'phone (72%). CONCLUSIONS: Patient satisfaction with family practice care in Slovenia was shown to be relatively high and can be compared to other European countries. The results showed areas in which quality improvement is required: organizational changes to shorten the waiting time in the waiting room and greater emphasis on communication skills. PMID- 10830672 TI - A systematic review of the content and quality of wrist outcome instruments. AB - OBJECTIVE: To assess the content and quality of published wrist outcome instruments using standardized criteria. DESIGN: An analytical study that examined 32 wrist outcome instruments sourced from textbooks, Medline (1951 to present) and Current Contents. MAIN MEASURES: The content of each instrument was classified into four categories: traditional measures (such as range of movement and strength), measures of the ability to perform daily activities, compensatory mechanisms used, and 'other'. Analysis included the frequency of assessment per category and the method of assessment. In addition, each instrument was graded using 13 quality criteria. Three criteria (scientific justification of the content and scoring system used, demographic utility) were considered to be essential. RESULTS: Eighty-two per cent of instruments reviewed for this paper contained traditional measures, of which most were assessed objectively. The ability to perform specific daily activities was assessed in 31% of the instruments whereas compensatory mechanisms were evaluated in only one instrument. These variables were not assessed in a consistent manner. Using the quality scoring system derived for this study, the quality of the instruments was generally poor. Only one instrument fulfilled all of the essential criteria. Only four instruments completely satisfied more than 50% of the criteria. CONCLUSIONS: Most wrist outcome instruments neglected to assess the impact of the disorder on the individual. Outcome was generally not expressed in functional terms or in terms that were relevant to each individual. The majority of the reviewed articles had poor quality. Thus use of these instruments may preclude sensitive evaluation of the efficacy of any intervention. PMID- 10830673 TI - Preventing medication errors. PMID- 10830674 TI - A comparison of 12- and 15-lead ECGS in ED chest pain patients: impact on diagnosis, therapy, and disposition. AB - The objective of this study was to investigate the diagnostic and therapeutic impact of the 15-lead electrocardiogram (15ECG) on the emergency department (ED) management of chest pain (CP) patients. The design was prospective use of 15ECG with real-time physician survey and retrospective comparison to 12-lead ECG (12ECG). The study took place in a University hospital ED. Adult CP patients participated. During the 15ECG period (June 1996 to July 1996), 595 patients (92% of CP patients) had 15ECG analysis. Diagnoses of acute coronary ischemic syndromes (ACIS) were as follows: 13 acute myocardial infarction (AMI, 7 anterior [ANT], 5 inferior [INF], 1 lateral [LAT], 2 posterior [POST], 1 right ventricular [RV]) and 136 unstable angina (USA) with 47% exhibiting ECG abnormality; the 2 POST and 1 RV AMI occurred in the setting of coexisting INF AMI. The following management strategies were used: 6 fibrinolytic therapy (TT), 4 primary angioplasty (PTCA), 67 rule-out myocardial infarction (ROMI), and 144 admission to critical care unit (CCU). During the 12ECG period (June 1995 to July 1995), 599 patients were encountered. The diagnoses of ACIS were as follows: 11 AMI (5 ANT, 4 INF, 2 LAT) and 146 USA with 51% exhibiting ECG abnormality (P = NS for diagnostic comparisons to 15ECG). The following management strategies were used: 5 TT, 5 PTCA, 59 ROMI, and 137 admission to CCU (P = NS for all treatment comparisons to 15ECG). Of 15ECG cases 81% had completed real-time physician survey, showing that the diagnosis and management ACIS were not altered by the 15ECG; physicians felt, however, that the 15ECG provided a more complete anatomic picture of the ACIS. No false-positive cases of additional lead STE were noted in this investigation except in cases involving abnormal intraventricular conduction such as the bundle branch block scenario. The 15ECG provided a more complete description of myocardial injury without altering the ED diagnosis, ED-based therapy, or hospital disposition in adult CP patients. Further study is required to identify patient subset(s) which may benefit from the 15ECG. PMID- 10830675 TI - ED evaluation of the pediatric trauma patient by ultrasonography. AB - The purpose of this study was to determine the accuracy of ultrasound examination of pediatric trauma patients by emergency physicians. Pediatric (age less than 18 years) trauma patients presenting to the emergency department of a level I trauma center were prospectively examined with bedside ultrasound during the secondary survey of their trauma resuscitation. Examinations were performed by emergency medicine residents and attending physicians who had completed an 8-hour course on trauma ultrasonography. Trauma physicians providing care to the patient were blinded to the results of the examination. In 47 children (median age 9 years) computed tomography of the abdomen/pelvis or laparotomy were also performed and served as gold standards to verify the presence or absence of free fluid in the abdomen. Sensitivity, specificity, and accuracy of the ultrasound examination for the detection of free fluid in the abdominal cavity was 75% (95% confidence interval [CI] 36% to 95%), 97% (95% CI 81% to 100%), and 92% (95% Cl 77% to 98%). Positive and negative predictive values were 90% (95% CI 46% to 100%) and 92% (95% CI 74% to 99%), respectively. Ultrasound examinations took an average of 7 minutes and 36 seconds, although this did not take into consideration delays created by interruptions for other diagnostic tests or procedures. An emergency physician and radiologist agreed on blinded interpretations of 83% of the examinations (kappa = 0.56). Bedside ultrasonography is a reliable and rapid method for screening traumatized children for the presence or absence of free fluid in the peritoneum even in the hands of novice sonographers. PMID- 10830676 TI - Analgesic administration to patients with an acute abdomen: a survey of emergency medicine physicians. AB - The objective of this study was to examine current practice patterns of analgesia administration among emergency physicians (EPs) when caring for a patient with an acute abdomen. Cross sectional data were acquired by a survey mailed in October 1997 to 1,000 American College of Emergency Physicians (ACEP) members from a purchased ACEP mailing list which contained 1,000 randomized ACEP members. A repeat survey was sent to nonresponders 2 months later and a random subset of recurrent nonresponders were telephoned. The questionnaire focused on physician demographics, practice patterns, and factors which influenced physician decision of when and whether to administer pain medication, specifically opiates, to patients with an acute abdomen. Forty-four percent of surveys were returned. Seventy-seven percent of respondents were men with an average of 10 years of experience. Fifty-seven percent were residency trained in emergency medicine. Although eighty-five percent felt that the conservative administration of pain medication did not change important physical findings on the physical examination, 76% choose not to give an opiate analgesic until after the examination by a surgeon. Twenty-five percent of patients did not receive any pain medication in the department. In conclusion, although EPs report that the judicious administration of pain medication does not mask important examination findings, the majority wait until after the surgeon has evaluated the patient to deliver analgesics. PMID- 10830677 TI - Underestimation of case severity by emergency department patients: implications for managed care. AB - The objective was to examine differences in symptom severity assessment by emergency department (ED) patients and by emergency physicians (EPs) and to relate these assessments with case management and disposition. The design was prospective convenience sample of ED patients. The setting was a U.S. university hospital ED with an annual ED patient census 28,000. The participants were all ED patients registered when first author was in ED; excluded were patients treated by the major trauma response team and those with a psychiatric chief complaint. All patients were interviewed by the first author and asked to classify their symptoms as emergent, urgent, or nonurgent; the EP attending classed patients' symptoms at presentation and after work-up was complete. Three hundred-one cases were entered in the study from May to August 1996. Although 28% of ED patients self-rated their symptoms as nonurgent, 5% of this group required hospital admission. Of this group 35% were assessed by the EP attending as having required emergent or urgent ED care. Of this group 5% also rated by the EP initially as nonurgent had their case severity upgraded after work-up. Reliance on either patient symptom self-assessment or physician screening assessment by telephone to determine appropriateness of an ED visit is not reliably safe for at least 5% of presenting patients. Even prospective ED visit severity assessment does not reliably identify "unnecessary" ED visits. PMID- 10830678 TI - Lack of t-PA use for acute ischemic stroke in a community hospital: high incidence of exclusion criteria. AB - Thrombolytic therapy with t-PA for acute ischemic stroke may provide benefit in long-term outcome. This retrospective study was undertaken to evaluate appropriateness of the National Institute of Neurological Disorders and Stroke (NINDS) protocol in the emergency department (ED). All patients with appropriate International Classification of Diseases, 9th revision (ICD-9) codes indicating stroke who presented to our 387-bed trauma-I community hospital during 1997 were included in the study. Of the nearly 35,000 patients screened, 201 patients satisfied our inclusion criteria. Mean age was 73.5 +/- 13.3 years. Men were evaluated and transported to computed tomography more rapidly and older patients more slowly. Nonwhites were more likely to arrive via emergency medical services (EMS). Average time from EMS arrival at scene to ED arrival was 22.7 minutes, and from ED arrival to triage was 8.4 minutes. The most common reason for exclusion from t-PA administration was delayed presentation (n = 188); this is the most serious barrier to use of t-PA for acute ischemic stroke. Extensive public education may combat this. PMID- 10830679 TI - The efficacy of cyanoacrylate-derived surgical adhesive for use in the repair of lacerations during competitive athletics. AB - Octylcyanoacrylate (Dermabond) is approved by the Food and Drug Administration for laceration closure. International studies have shown its utility in wound closure and have shown it to be as good or better than suture closure for speed, patient preference, and cosmesis, with no difference in the rate of dehiscence or infection. We sought to determine whether it retains its tensile strength, durability, and skin apposition when an athlete is allowed to reenter competition, where it is subject to recurrent stress, moisture, and trauma. The study was performed at two professional hockey sites. Wounds were anesthetized, irrigated, and debrided. The skin was closed with Dermabond. The athlete was returned immediately to competition. Wounds were examined at the end of competition and again at 7 days. A total of 32 lacerations on 28 players were studied. The mean size of laceration was 2.3 cm (range 0.8 cm to 4.5 cm). The majority (95%) of wounds were on the face. Of the 32 lacerations, 31 (97.6%) had good results at the conclusion of the game. Of these 31, all had good results at 7 days following repair. Dermabond retained its strength, durability, and skin apposition when the athlete was allowed to reenter competition following wound repair. PMID- 10830680 TI - Hyponatremia in the emergency department. AB - Hyponatremia is a clinical manifestation of a wide variety of diseases, some of which have high mortality rates. To assess the prevalence, cause, and prognosis of hyponatremia encountered in the emergency department, we conducted a prospective study at a major hospital in southern Taiwan. We included all adult internal medicine patients treated in the emergency department during a 2-month period. Hyponatremia was defined as a serum sodium level below 134 mEq/L, and cases patients were followed till being discharged. Among the 3,784 patients included, 166 case patients were identified. Most (65%) case patients were hypovolemic, and the overall mortality rate was 17.9%. The mortality rate increased as the sodium level decreased, but was not related to gender, age, cause, or serum potassium level. When 21 hyperglycemic patients whose serum sodium levels went beyond 134 mEq/L after the adjustment for blood sugar levels were excluded, the prevalence of true hyponatremia was 3.83%. The most common underlying diseases were those of the gastrointestinal system. It is concluded that hyponatremia is a common condition encountered in the emergency department. The mortality is correlated with the serum sodium level, and adjustment of the level is required in hyperglycemic patients to make a correct diagnosis. Unlike the cases in some other clinical settings, almost all cases of hyponatremia encountered in the emergency department were not iatrogenic and had recognizable underlying diseases. Therefore, more effort is generally required to identify the cause of hyponatremia cases in the emergency department. PMID- 10830681 TI - Risks of intravenous administration of hypotonic fluids for pediatric patients in ED and prehospital settings: let's remove the handle from the pump. AB - Fluid resuscitation of infants and children is a common management problem in prehospital and emergency department care. We present two cases of children who received 5% dextrose in water as the initial resuscitation fluid. Bolus administration of hypotonic fluid contributed to fatal outcomes in these cases. Recommendations are made for eliminating hypotonic fluids as stock items in both the prehospital and emergency department settings. PMID- 10830682 TI - Corneal abrasions associated with pepper spray exposure. AB - Pepper spray containing oleoresin capsicum is used by law enforcement and the public as a form of nonlethal deterrent. Stimulated by the identification of a case of a corneal abrasion associated with pepper spray exposure, a descriptive retrospective review of a physician-maintained log of patients presenting to a jail ward emergency area over a 3-year period was performed. The objective was to give some quantification to the frequency with which an emergency physician could expect to see corneal abrasions associated with pepper spray exposure. Of 100 cases of pepper spray exposure identified, seven patients had sustained corneal abrasions. We conclude that corneal abrasions are not rare events when patients are exposed to pepper spray and that fluorescein staining and slit lamp or Wood's lamp examination should be performed on all exposed patients in whom corneal abrasions cannot be excluded on clinical grounds. PMID- 10830683 TI - The stability of arterial blood gases during transportation of patients using the RespirTech PRO. AB - The transportation of critically ill patients requiring mechanical ventilation is recognized as a high-risk and expensive procedure. Approaches have included using manual bag-type valve resuscitators and expensive portable transport ventilators. This study evaluated the effectiveness of the inexpensive portable RespirTech PRO (RTP) gas-powered automatic resuscitator during intrahospital transport of critically ill mechanically ventilated patients. Twenty medical intensive care patients on stable mechanical ventilator settings had arterial blood gas and vital sign determination before routine transport out of the intensive care unit (ICU). Repeat measurements were made during transport approximately 30 minutes after being placed on the RTP portable pressure-cycled automatic resuscitator using an FiO2 of 100%. During use of the RTP for transport, there were no statistically significant variations observed in mean arterial blood pressure [82 +/- 11 SD (range 65 to 112) mm Hg before transport versus 85 +/- 14 SD (range 59 to 110) mm Hg during transport], heart rate [94 +/- 16 SD (range 74 to 127) beats/min) before versus 96 +/- 17 SD (range 69 to 132) beats/min during], arterial pH [7.41 +/- 0.07 SD (range 7.31 to 7.58) before versus 7.42 +/- 0.05 SD (range 7.37 to 7.52) during], and PaCO2 [43 +/- 10 SD (range 26 to 65) mm Hg before versus 43 +/- 10 SD (range 27 to 61 mm Hg) during]. Because the FiO2 before transport was 63 +/- 26 SD (range 30% to 100%) versus 100% during transport using the RTP, the mean PaO2 was significantly increased from 124 +/- 86 SD (range 57 to 367) mm Hg before transport to 297 +/- 168 SD (range 65 to 537) mm Hg during (P< .001). No transportation associated clinical adverse events were noted. Several previous investigations have shown that portable ventilators are safe and effective in intrahospital transport of mechanically ventilated patients. This study showed that the portable pressure-cycled RTP also allows safe transportation of mechanically ventilated ICU patients. By analogy, the RTP is potentially useful as an automatic resuscitator for emergency medical care. This RTP is a disposable resuscitator/ventilator device that provides an inexpensive alternative for transporting ventilator-dependent patients. PMID- 10830684 TI - Patient perceptions of the specialty of emergency medicine. AB - The study objective was to determine emergency department (ED) patients' perceptions of the specialty of emergency medicine. We surveyed a convenience sample of adult ED patients regarding their knowledge of the specialty of emergency medicine. Responses included: 22% believing that ED physicians have their own practice outside the ED; 26% of patients with primary care physicians expected to be seen by their primary care physician in the ED; 19% thought ED physicians care for patients after admission; 26% thought that ED physicians perform surgery, 62% perceived emergency medicine to be a specialty; 15% have heard of the American College of Emergency Physicians; 71% thought that ED physicians are board certified and 15% thought paramedics were ED physicians. Patients estimated ED physicians' mean annual mean salary to be $100,000 and 61% believe that ED physicians are hospital employees. In conclusion, the specialty of emergency medicine is not well understood by our patients. PMID- 10830685 TI - HIV postexposure prophylaxis for children and adolescents. AB - HIV postexposure prophylaxis (PEP) is now a well-established part of the management of health care workers after occupational exposures to HIV. Use of PEP for adults exposed to HIV after sexual contact or injection drug use in nonoccupational settings remains controversial with limited data available. There is even less information available concerning HIV PEP for children and adolescents after accidental needlestick injuries or sexual assault. The objective was to describe the current practice of and associated problems with HIV PEP for children and adolescents at an urban academic pediatric emergency department. A retrospective review of all children and adolescents offered HIV PEP between June 1997-June 1998 was conducted. Ten pediatric and adolescent patients were offered HIV PEP, six patients after sexual assault, four patients after needle stick injuries. There were two small children 2 and 3 years of age and eight adolescents. Of these 10 patients, eight were started on HIV PEP. The regimens used for PEP varied; zidovudine, lamivudine, and indinavir were prescribed for in seven patients and zidovudine, lamivudine, and nelfinavir for one other. All 10 patients were HIV negative by serology at baseline testing and all available for follow-up testing (5 of 10) remained HIV negative at 4 to 28 weeks. Only two patients completed the full course of 4 weeks of antiretroviral therapy. Financial concerns, side effects, additional psychiatric and substance abuse issues as well as the degree of parental involvement influenced whether PEP and clinical follow-up was completed. HIV PEP in the nonoccupational setting for children and adolescents presents a medical and management challenge, and requires a coordinated effort at the initial presentation to the health care system and at follow-up. The difficulties encountered in the patients in our series need to be considered before initiating prophylaxis. A provisional management approach to HIV PEP in children and adolescents is proposed. PMID- 10830686 TI - Autoimmune blistering skin diseases. AB - Emergency physicians, at the front line of patient care, are often confronted with a wide variety of dermatologic conditions. Prompt recognition is essential, especially for the autoimmune blistering skin diseases, many of which have considerable morbidity and mortality. Therefore, an accurate diagnosis is imperative for appropriate referral and initiation of therapy. This review article provides a concise yet thorough discussion of the clinical presentation, incidence, differential diagnosis and management of the commonly encountered autoimmune blistering skin diseases, some of which include pemphigus, bullous pemphigoid, and epidermolysis bullosa acquisita. PMID- 10830687 TI - The occupational hazards of emergency physicians. AB - Emergency physicians are exposed to a variety of occupational hazards. Among these are infectious diseases, such the human immunodeficiency virus, hepatitis B and C viruses, and tuberculosis. Hepatitis G virus is transmissible but may not be a cause of illness. The likelihood of being exposed to these agents appears to be higher in the ED than other medical settings but estimates of the prevalence of these diseases in the ED vary, depending on the patient population served. Estimates of risk for contracting these infections are reviewed. Measures to prevent these exposures can reduce risk, but compliance is low, particularly for those involving changes in the behavior of emergency physicians (such as not recapping needles). Latex allergy is a hazard of health care workers. Its prevalence is reported to be quite high, but these findings are difficult to interpret in the absence of a universally accepted definition of the condition. Its prevalence in emergency physicians is not known. Other noninfectious hazards include workplace violence and exposure to nitrous oxide. The health effects of rotating shift work may put emergency physicians at increased risk of coronary artery disease and impaired reproductive health. Emotional stress is another hazard of emergency physicians, and may lead to burnout. PMID- 10830688 TI - Artifactual electrocardiographic change mimicking clinical abnormality on the ECG. AB - Electrocardiographic artifact is a common finding in patients requiring evaluation and monitoring in the prehospital, emergency department, or intensive care unit settings. Artifact results from both internal (physiological) and external (nonphysiological) sources. In most instances, artifact is recognized as an incorrect electrocardiographic signal--its only impact producing interference in electrocardiogram interpretation; artifact may also produce electrocardiographic signals which mimick disease--these signals the physician must recognize as artifact. PMID- 10830689 TI - Luxatio erecta: a rarely seen, but often missed shoulder dislocation. AB - Luxatio erecta is an uncommon disorder and presents in a unique, unusual manner. Luxatio erecta is often misdiagnosed as an anterior dislocation. The presentation is unmistakable and classic: the arm hyperabducted and locked above the head. Neurovascular injuries consist of neuropraxia of brachial plexus, radial and ulnar nerve. Vascular injuries are complicated by intimal tears, transections, and/or thrombosis of the axillary artery or vein. Reduction is done with the traction and countertraction maneuver. Once it is reduced the arm is then placed and maintained with a sling in adduction to the chest. Orthopedic referral is required because of the high incidence of rotator cuff injury. PMID- 10830690 TI - Cases in electrocardiography. PMID- 10830691 TI - Renal infarction: an uncommon mimic presenting with flank pain. AB - A 39-year-old woman arrived to the emergency department complaining of a constant, progressive, left flank pain, with no beneficial effect from spasmolytic and nonsteroidal antiinflammatory drugs. Two years before, she suffered another episode of right flank pain and stranguria, but instrumental examinations (ultrasonography, urography) remained negative. Besides a mild tenderness in the left flank, physical examination was normal. Blood chemistry panel showed leukocytosis (17.2 x 10(3) mL, neutrophils 82.8%) and a slight increase of serum lactate dehydrogenase (LDH) (543 U/L versus 230 to 460 U/L). Urinanalysis showed a slight hemoglobinuria (0.5 mg/dL), and sediment contained some red cells and leukocytes. Diagnostic examinations (ultrasonography, computed tomography) showed a left renal nonhomogeneous space-occupying lesion, orientative for renal malignancy. She was transferred to the urology department and operated. Both intraoperatory and histological diagnosis was ischemic infarction and, after exclusion of all possible underlying causes, final diagnosis was idiopathic renal infarction. Diagnostic procedures and literature reports are discussed. PMID- 10830692 TI - Emergency department presentation of pituitary apoplexy. AB - Pituitary apoplexy is an acute infarction of pituitary gland, and potentially life-threatening condition that may be highly variable in its clinical presentation. We report a 54-year-old man presenting to the emergency department with an isolated oculomotor nerve palsy. Computed tomography (CT) scan revealed an isodense mass within sellar region and subsequently, magnetic resonance imaging (MRI) revealed a pituitary apoplexy causing a compression of right oculomotor nerve. The patient received hydrocortisone immediately, and did well with medical management. An isolated oculomotor nerve palsy is very rarely the presenting sign of pituitary apoplexy. When correctly diagnosed and treated, the third nerve palsy appears to be reversible. A pathophysiology, differential diagnosis, and treatment is described. PMID- 10830694 TI - Severe pulmonary complications after silicone fluid injection. PMID- 10830693 TI - Emergency medical services development in the Seychelles Islands. AB - The Republic of the Seychelles enjoys a sophisticated health care system by developing country standards. Basic food, water, sanitation, and preventative health care needs have been addressed and the Seychelles Ministry of Health has recently focused on the need to improve the country's emergency medical services (EMS). In response to this need, a joint international collaborative effort designed to improve the island's EMS was launched. This report profiles the current health care system in the Seychelles, with special emphasis on EMS. It also describes an international collaborative effort between the Seychelles Ministry of Health and the UCLA Center for International Emergency Medicine designed to improve existing EMS. PMID- 10830695 TI - Subglottic granuloma: a rare cause of difficulty breathing in children. PMID- 10830696 TI - Impending cardiac tamponade, an unseen danger? PMID- 10830697 TI - Survey of staff opinions on ultrasound by emergency physicians. PMID- 10830698 TI - Physical examination in determining the outcomes of ultrasound for DVT. PMID- 10830699 TI - Prognostic factors in acute cardiogenic pulmonary edema. PMID- 10830700 TI - Phenylpropanolamine-associated intracranial hemorrhage in an infant. PMID- 10830701 TI - Pencil stab wound results in pneumonia and pleural effusion. PMID- 10830702 TI - Steroid-induced mediastinal lipomatosis with radiographic features of pericardial effusion. PMID- 10830703 TI - High serum soluble Fas-ligand in cardiopulmonary arrest patients. PMID- 10830704 TI - Situs inversus: emergency evaluations of atypical presentations. PMID- 10830705 TI - Restoration of spontaneous circulation from asystole with aminophylline. PMID- 10830706 TI - Are pediatric ED physicians blowing off peak expiratory flows? PMID- 10830707 TI - Acute myocardial infarction associated wiht Sildenafil (Viagra) ingestion. PMID- 10830708 TI - Unrecognized right endobronchial intubation causing total left lung collapse in a pediatric trauma patient. PMID- 10830709 TI - Musculoskeletal training: emergency medicine needs to take charge. PMID- 10830710 TI - Accidental hypothermia in the subtropics. PMID- 10830711 TI - Revivarant (gamma-butyrolactone) poisoning. PMID- 10830712 TI - Transduction efficacy, antitumoral effect, and toxicity of adenovirus-mediated herpes simplex virus thymidine kinase/ ganciclovir therapy of hepatocellular carcinoma: the woodchuck animal model. AB - Gene therapy for hepatocellular carcinoma (HCC) has shown some promise, but its evaluation requires relevant experimental models. With this aim, we present an evaluation of the interest of using the woodchuck model of HCC to assess in vivo gene transfer efficiency. We tested the transduction efficacy of the adenoviral vectors directing lacZ gene product expression under the control of the cytomegalovirus and alpha-fetoprotein (AFP) regulatory sequences. We have also investigated whether an adenoviral cytomegalovirus-thymidine kinase (Tk) vector might induce an antitumoral effect in this model. Our results demonstrate that with direct intratumoral and intrahepatic arterial injections, transduction of a significant proportion of tumor cells occurred even in large HCC nodules. Furthermore, due to intra-arterial anastomoses, direct intratumoral injection led to transduction of some noninjected HCC nodules. Moreover, direct intratumoral injection of a herpes simplex virus-1 Tk-encoding vector induced, on ganciclovir administration, a significant antitumoral effect in the two animals evaluated. However, in one animal, massive hepatic failure occurred due to Tk expression in nontumor cells. These results emphasize the need to target the expression of the Tk gene to tumor cells using a hepatoma-specific promoter such as AFP promoter. However, we showed that, in vivo, lacZ expression as driven by the AFP promoter was extremely low, thus emphasizing some potential pitfalls when using this approach. Altogether, our data stress the need to test gene therapy-based strategies in such in vivo animal models of HCC and evaluate gene transduction, expression, and biological activity, as well as its potential toxicity. PMID- 10830713 TI - Replication-restricted vaccinia as a cytokine gene therapy vector in cancer: persistent transgene expression despite antibody generation. AB - BACKGROUND: As antitumoral immunity requires the generation of local immunity directed against tissue proteins, we attempted to recreate within tumors the same environment found within tissues affected by autoimmune diseases (i.e., prolonged cytokine expression). Vaccinia virus (VV) has not been widely used as a cytokine gene therapy vector because of presumed high immunogenicity that would likely make repeated injections impossible; therefore, we modified it by inserting the cytokine gene into the thymidine kinase region, rendering it replication restricted. The cytokine chosen was human interleukin-2 (IL-2); a molecule with powerful antitumoral effects. METHODS: Six patients with the treatment-resistant tumor malignant mesothelioma received intratumoral (i.t.) VV-IL-2 therapy for 12 weeks by injection of 10(7) plaque-forming units of VV-IL-2 per dose. Serial tumor biopsies, sputum, urine, and blood samples were tested for VV-IL-2 mRNA expression; VV culture and T-cell infiltrates were evaluated by immunohistochemistry. Patients and contacts of patients were monitored for changes in VV immunoglobulin G (IgG) levels and clinical evidence of VV infection. RESULTS: VV-IL-2 was not excreted and was only cultured in one patient from tumor biopsies. A T-cell infiltrate was detected in 50% of tumor biopsies. VV-IL-2 mRNA expression was highest on days 1-3 postinjection and was detected for up to 3 weeks after each injection even though VV IgG levels rose in all patients. No significant toxicities, infection of patient contacts, or tumor regressions were observed. CONCLUSIONS: I.t. VV-IL-2 administration is safe, is associated with minimal toxicity, and results in i.t. expression of VV-IL-2 for up to 3 weeks postinjection regardless of the level of anti-VV IgG titers generated. This suggests that VV may be a good vector for repeated cytokine gene therapy of solid human cancer. PMID- 10830714 TI - Ribozyme-mediated suppression of platelet type 12 lipoxygenase in human erythroleukemia cells. AB - The platelet type 12 lipoxygenase (12-LOX) adds molecular oxygen to C-12 arachidonic acid to yield 12-hydroperoxy-5,8,10,14-eicosatetraenoic acid. It has been suggested that 12-LOX and its metabolites play an important role in tumor angiogenesis. A hammerhead ribozyme (Rz) targeted to the first GUC site within the 12-LOX mRNA was designed and cloned into an in vitro transcriptional or mammalian expression vector. In vitro, the Rz was able to cleave its substrate efficiently in a time-dependent manner. Under multiple turnover conditions, the Rz performed well at 37 degrees C, with a further improvement at 50 degrees C. When cloned into a mammalian expression vector, pSV2neo, the Rz construct efficiently decreased the level of 12-LOX mRNA in stably transfected human erythroleukemia cells to levels that were undetectable by Northern blot analyses. 12-LOX enzyme activity assays showed that Rz significantly reduced the 12 hydroperoxy-5,8,10,14-eicosatetraenoic acid production in human erythroleukemia cells; this effect was sustained for up to 6 months in cell culture. The Rz developed in this study may represent a powerful tool for potential applications, ranging from an understanding of tumor angiogenesis to cancer gene therapy. PMID- 10830715 TI - Antitumoral vaccination with granulocyte-macrophage colony-stimulating factor or interleukin-12-expressing DHD/K12 colon adenocarcinoma cells. AB - Immunomodulating gene therapy for the treatment of malignant diseases is under extensive investigation. In this study, we induced an antitumoral immune response with murine interleukin-12 (mIL-12) and murine granulocyte-macrophage colony stimulating factor (GM-CSF)-secreting tumor cells in a model of peritoneal carcinomatosis. Intraperitoneal injection of DHD/K12 tumoral cells engineered to produce IL-12 or GM-CSF did not generate any tumors, whereas untransduced DHD/K12 cells gave rise to peritoneal carcinomatosis. IL-12-expressing DHD/K12 cells also protected against tumors derived from coinjected parental cells. To test whether cytokine-producing cells could elicit a memory antitumoral immune response, animals received a challenge with parental DHD/K12 cells 35 days after the injection of proliferating or irradiated DHD/K12 engineered cells. Under our experimental conditions, irradiated tumor cells did not generate any antitumoral immunity. In contrast, tumor development was delayed and survival increased in the animals vaccinated with cytokine-secreting proliferating cells. A specific cytotoxic T-lymphocyte response against DHD/K12 parental cells was observed after vaccination with GM-CSF-expressing cells. Our results demonstrated that intraperitoneal vaccination with IL-12- or GM-CSF-expressing adenocarcinoma cells induced a systemic immune antitumoral response that may be useful as an adjuvant therapy after surgical resection of colorectal cancer. PMID- 10830716 TI - Development of cationic liposome formulations for intratracheal gene therapy of early lung cancer. AB - Regional (intratracheal or aerosol) delivery of cationic liposome-DNA complexes for gene therapy of lung disease offers distinct advantages over systemic (intravenous) administration. However, optimal formulations for early lung cancer treatment have not been established. Therefore, we investigated >50 different liposome and micelle formulations for factors that may affect their transcription efficiency and tested the ideal formulations in an in vivo mouse model. Our data showed that cationic liposomes were generally more effective at transfecting genes than were micelles of the same lipid composition, thus suggesting a role for the bilayer structure in facilitating transfection. In addition, the transfection efficiency of liposome-delivered genes was highly dependent upon the lipid composition, lipid/DNA ratio, particle size of the liposome-DNA complex, and cell lines used. By optimizing these factors in vitro and in vivo, we developed a novel liposome formulation (DP3) suitable for intratracheal administration. Using G67 liposome as control, we found that DP3 was more effective than G67 in vitro and as effective as G67 at both preventing lung tumor growth and prolonging survival in our lung cancer mouse model. We observed a positive correlation between the in vitro p53 function and the in vivo antitumoral activities of liposome-p53 formulations, which had not been reported previously in studies of an intravenous liposome gene delivery system. This correlation may facilitate the development and optimization of a liposome-p53 formulation for aerosol use in lung cancer patients. PMID- 10830717 TI - Retroviral gene transfer of dominant negative raf-1 mutants suppresses ha-ras induced transformation and delays tumor formation. AB - Activating mutants of ras are among the most frequently found genetic alterations in human cancers. Therefore, Ras appears to be an attractive target for therapeutic intervention using gene transfer. The protein kinase Raf-1 acts as a direct downstream effector of Ras and is involved in Ras-induced cellular transformation. Using the NIH3T3 fibroblast-derived tumor cell line PEJ, which expresses oncogenic Ha-rasG12V, we analyzed whether dominant negative mutants of Raf-1 can inhibit Ras-mediated transformation. Retroviral gene transfer was used to stably transduce PEJ cells with three different dominant negative mutants of Raf-1. This resulted in reversion of the transformed phenotype in vitro as evidenced by an increase in contact inhibition and reduced anchorage-independent growth. However, tumor formation in nude mice was significantly delayed only by one of these mutants. Therefore, dominant negative mutants of the oncoprotein Myc, which is known to synergize with Raf-1 in tumor formation, were transduced into PEJ cells expressing a dominant negative Raf mutant. This leads to killing of the cells. These results indicate that although interference with Ras-induced transformation using dominant negative mutants of Raf is feasible and effective in vitro using retroviral vectors, an additional block (e.g., that of Myc) is necessary to kill PEJ cells. These results also indicate that interference with Ras-dependent signaling is not sufficient for inhibition of tumor formation of PEJ cells in vivo. PMID- 10830718 TI - Antitumoral effect of interleukin-12-secreting fibroblasts in a mouse model of ovarian cancer: implications for the use of ovarian cancer biopsy-derived fibroblasts as a vehicle for regional gene therapy. AB - Fibroblasts can easily be cultured from human solid ovarian tumor biopsies and are readily transfected using retroviral vectors. To test the feasibility of using these cells as a vehicle for regional, intraperitoneal (i.p.) ovarian cancer gene therapy, we first examined the behavior of murine fibroblasts transduced with the neoR marker gene and injected i.p. in syngeneic mice. We found that these fibroblasts were not invasive and formed clusters preferentially attached to the pancreatic and hepatic mesentery, where they survived for at least 30 days and continued to express neoR. We subsequently assessed the effect of regional delivery of murine interleukin-12 (mIL-12)-secreting fibroblasts in a syngeneic immunocompetent mouse model of ovarian cancer (ID8). ID8 ovarian tumor cells were injected i.p. into one flank. Our results show a survival of 88% at 120 days postinjection of animals treated with mIL-12-secreting fibroblasts i.p. in the other flank on the same day as injection of tumor cells. At that time, all animals coinoculated with ID8 cells and unmodified fibroblasts had been or needed to be culled because of advanced neoplastic disease (P < 10(-5), log-rank test). Furthermore, animals treated with mIL-12-secreting fibroblasts had a statistically significant decrease in tumor burden, as measured by diaphragm weight, relative to mock-treated groups (P = .0032, H-test). Our histological data show evidence for both immunological and anti-angiogenic mechanisms being implicated in this antitumoral effect. In addition, no signs of IL-12-induced toxicity were observed in any of the tissue sections analyzed. Taken together, our findings suggest that ovarian tumor biopsy-derived fibroblasts are a safe and efficacious vehicle for i.p. delivery of IL-12 as a regional secondary treatment for minimal residual disease of ovarian cancer. PMID- 10830719 TI - Expression of Escherichia coli B nitroreductase in established human tumor xenografts in mice results in potent antitumoral and bystander effects upon systemic administration of the prodrug CB1954. AB - Expression of the Escherichia coli enzyme nitroreductase (NTR) in mammalian cells enables them to activate the prodrug 5-(aziridin-1-yl)-2,4-dinitrobenzamide (CB1954), leading to interstrand DNA cross-linking and apoptosis in both proliferating and quiescent cells. In the work reported here, we used human hepatocellular carcinoma and squamous carcinoma cell lines constitutively expressing NTR to demonstrate that the ntr/CB1954 system results in potent, long lasting antitumoral effects in mice. We also demonstrate that this enzyme/prodrug combination results in antitumoral effects in vivo when only a minority of tumor cells express the enzyme, using either cells constitutively expressing NTR or ntr gene delivery in situ. PMID- 10830720 TI - Adenovirus-mediated transfer of p53 and Fas ligand drastically enhances apoptosis in gliomas. AB - Various therapeutic approaches toward killing glioma cells by inducing apoptosis have been developed, but these approaches are often hampered by anti-apoptotic mechanisms. In this study, we attempted to develop a technique that overrides the resistance toward apoptosis in glioma cells. To date, p53- and Fas-mediated apoptotic pathways have been shown to be different. Therefore, we carried out a gene therapy that combines the pro-apoptotic effect of these two different pathways. The recombinant adenoviruses (Advs) for p53 and Fas ligand (FL) (Adv p53 and Adv-FL, respectively) were constructed. Transfecting the p53 gene into glioma cell lines (A-172 and U251 glioma cells) led to overexpression of Bax, a protein that induces permeability transition; at the same time, this transfection brought about an overexpression of Fas. To intensify Fas-mediated apoptosis, we transferred the FL gene together with the p53 gene by Adv-mediated gene transduction into A-172 and U251 cells. Coinfecting Adv-p53 and Adv-FL into A-172 cells, which are relatively resistant to apoptosis by infection with Adv-p53 or Adv-FL alone (Adv-p53, multiplicity of infection (MOI) of 100: 8.5 +/- 0.7%; Adv FL, MOI of 100: 3.0 +/- 0.1%) resulted in a drastic enhancement of the percentage of apoptotic cells (Adv-p53 and Adv-FL, each at an MOI of 30: 24.2 +/- 0.9%). Coinfection with Adv-p53 and Adv-FL in U251 cells resulted in a similar enhancement of the percentage of apoptotic cells (Adv-p53 and Adv-FL, each at an MOI of 30: 59.0 +/- 2.3%) compared with that seen in U251 cells transfected with Adv-p53 or Adv-FL alone (Adv-p53, MOI of 30: 3.1 +/- 0.3%; Adv-FL, MOI of 30: 18.1 +/- 0.3%). Regardless of whether a cell line is resistant or sensitive to FL and p53-mediated apoptosis, coinfection with Adv-p53 and Adv-FL dramatically enhanced the degree of apoptosis of glioma cells. Our results indicate that the coinfection approach can be used as a modality for the gene therapy of gliomas, overriding the apoptosis-resistant mechanisms in glioma cells. PMID- 10830721 TI - Adenovirus-mediated transfer of bax with caspase-8 controlled by myelin basic protein promoter exerts an enhanced cytotoxic effect in gliomas. AB - The transduction of Bax protein, which is up-regulated in radiation- and chemotherapy-induced apoptosis, augments the cytotoxicity of radiotherapy and chemotherapy for cancers. The cytotoxicity of Bax overexpression is caused primarily by mitochondrial dysfunction, which is also involved in the apoptosis triggered by caspase-8. In this study, we transduced the Bax gene in combination with caspase-8 gene to evaluate whether or not this approach induces effective cytotoxicity in glioma cells. In terms of cancer gene therapy, it is critically important to induce cytotoxic genes in a cancer-specific manner. Therefore, we used the myelin basic protein promoter to drive cytotoxic genes. The expression level controlled by the myelin basic protein promoter was relatively low in gliomas. In U251 and U-373 MG glioma cells, adenovirus-mediated transduction of the Bax gene combined with caspase-8 gene induced enhanced apoptosis and cell death as determined by morphological analysis and assay for dead cells, hypodiploid cells, and DNA fragmentation (terminal deoxynucleotidyltransferase mediated deoxyuridine triphosphate nick end labeling method). This therapeutic modality would be useful to induce a specific and enhanced cytotoxic effect for gliomas. PMID- 10830722 TI - Endogenous p53 gene status predicts the response of human squamous cell carcinomas to wild-type p53. AB - Prior reports suggest that p53 protein status may influence the response to gene transduction with wild-type (wt) p53. Adenoviral vectors containing the p53 gene were administered to normal keratinocytes, to squamous cell carcinoma (SCC) lines with varied p53 protein status (absent, mutant, wt, or degraded by papillomavirus), as well as to tumors formed in severe combined immunodeficient mice. The percentage of cells undergoing apoptosis, G1 growth arrest, WAF1/p21 induction, and in vivo tumor progression were studied after wt p53 gene transduction. Apoptosis developed first in normal keratinocytes, next in SCCs lacking p53 protein, and last in SCCs with mutant or degraded p53 protein. All of the cell lines studied demonstrated an increase in WAF1/p21 protein, but only those lacking p53 protein showed G1 arrest. Tumors lacking p53 protein were more susceptible to p53 overexpression than those containing mutant or degraded p53 protein. The endogenous p53 protein status of SCCs appears to influence the outcome of p53 gene transduction. PMID- 10830723 TI - Coexpression of rat glutathione S-transferase A3 and human cytidine deaminase by a bicistronic retroviral vector confers in vitro resistance to nitrogen mustards and cytosine arabinoside in murine fibroblasts. AB - The transfer of drug resistance genes into hematopoietic cells is an experimental approach to protect patients from drug-induced myelosuppression. Because anti cancer drugs are often administered in combination to increase their clinical efficacy, vectors that express two drug resistance genes are being developed to broaden the spectrum of chemoprotection. We have constructed a bicistronic vector, MFG/GST-IRES-CD (MFG/GIC) coexpressing rat glutathione S-transferase (GST) A3 isoform (rGST Yc1) and human cytidine deaminase (CD). Murine NIH 3T3 fibroblast cells transduced with this vector were evaluated for their resistance to nitrogen mustards and cytosine nucleoside analogs. GIC-transduced polyclonal cell populations (GIC cells) demonstrated marked increases in selenium independent glutathione peroxidase (peroxidase) and CD activities, as well as increased resistance to melphalan (2.3-fold), chlorambucil (3.4-fold), and cytosine arabinoside (Ara-C) (8.1-fold). After selection with Ara-C, the peroxidase and CD activities of GIC cells were augmented 2.6- and 2.9-fold, respectively, in comparison with unselected cells, and the resistance to melphalan, chlorambucil, and Ara-C was further increased to 3.7-, 5.9-, and 53 fold, respectively. Melphalan selection of GIC cells likewise augmented their peroxidase (2.3-fold) and CD (1.9-fold) activities. GIC cells proliferated in the simultaneous presence of melphalan and Ara-C at drug concentrations that completely inhibited the growth of untransduced cells. The growth rate of unselected GIC cells exposed to the drug combination averaged 18% that of drug free cultures. The growth rate of GIC cells exposed to the drug combination increased to 30% of controls after Ara-C selection and to 50% after melphalan selection. Our results suggest that retroviral transfer of MFG/GIC may be useful for chemoprotection against the toxicities of nitrogen mustards and cytosine nucleoside analogs. PMID- 10830724 TI - Preclinical study on gene therapy of cervical carcinoma using adeno-associated virus vectors. AB - Approximately 90% of cervical carcinomas are causally linked to infections with high-risk human papillomaviruses (HPVs), whose oncogenicity has been assigned to the continued expression of two early genes, E6 and E7. Reversal of the transformed phenotype by inhibiting E6/E7 gene expression therefore provides a suitable goal for future tumor therapy. Using recombinant adeno-associated virus type 2 (AAV-2) vectors, two types of therapeutic genes were expressed in cervical carcinoma cells with the aim of suppressing the E6/E7 oncogenes: (a) antisense E6/E7 and ribozyme genes and (b) the monocyte chemoattractant protein-1 (MCP-1) gene encoding MCP-1. Previous studies have shown that the MCP-1 protein is able to indirectly repress E6/E7 gene expression and is consistently absent in tumorigenic HPV-positive cervical carcinoma cell lines. Here, the effect of these therapeutic genes on tumor formation is analyzed in nude mice after ex vivo gene transfer into a HPV16- or HPV18-positive cervical carcinoma cell line (HeLa or SiHa, respectively). Whereas AAV-2 vector-mediated transfer of antisense or even ribozyme genes did not significantly influence tumor formation from implanted SiHa cells, the transfer and expression of human MCP-1 strongly inhibited the development of tumors derived from either HeLa or SiHa cells. Similar results were also obtained after in vivo delivery of these genes into SiHa-derived tumors. This suggests that transfer of therapeutic genes mediating a systemic effect via recombinant AAV-2 vectors offers a promising approach for the development of gene therapies directed against papillomavirus-induced human cancers. PMID- 10830725 TI - Radiosensitization of rat glioma with bromodeoxycytidine and adenovirus expressing herpes simplex virus-thymidine kinase delivered by slow, rate controlled positive pressure infusion. AB - Infection of rat RT2 glioma cells in vitro with an adenovirus (ADV-TK) expressing herpes simplex virus (HSV) thymidine kinase (TK) and subsequent exposure to 5 bromo-2'-deoxycytidine (BrdC), which is specifically incorporated into ADV-TK infected cell DNA as 5-bromo-2'-deoxyuridine (BrdU), results in significant radiosensitization (sensitizer enhancement ratio: 1.4-2.3) compared with Ad beta gal-infected cells. Cell killing correlated well with increased BrdU DNA incorporation and with apoptosis. Whereas radiation (4 Gy) alone was relatively ineffective in inducing apoptosis, treatment with HSV-TK/BrdC resulted in BrdC dose- (10-100 microM) and time-dependent (24-48 hours) increases, and the combination of the two treatments produced a synergistic response (1.5- to 2 fold). To investigate the effects of the ADV-TK/BrdC treatment in vivo, RT2 cells were grown as soft tissue tumors in Fischer 344 rats and conditions for virus infusion were optimized by altering the volume and rate of infusion using a rate controlled positive pressure device. We found that relatively large volumes (100 150 microL) of virus delivered at rates of < or = 1 microL/minute were optimal and gave uniform and reproducible results. Using these optimal infusion conditions, we were able to achieve 40% adenovirus infection in the tumor. Infection of RT2 tumors with ADV-TK and continuous administration of BrdC from an osmotic pump resulted in significant (.001 < P < .009) tumor regression 6 days after radiation (30 Gy delivered as 2 x 5 Gy over 3 days) compared with controls. In situ staining of sectioned tumors with anti-BrdU antibody or by high performance liquid chromatography analysis of extracted and hydrolyzed tumor DNA confirmed that we obtained efficient and specific incorporation of BrdU into tumor cells. These results suggest that adenovirus-mediated delivery of HSV-TK in combination with BrdC and radiation can potentially be an efficient combination modality for the treatment of gliomas. PMID- 10830726 TI - Cancer gene therapy mediated by CTS1, a p53 derivative: advantage over wild-type p53 in growth inhibition of human tumors overexpressing MDM2. AB - Recently, a new p53 derivative has been designed, namely chimeric tumor suppressor 1 (CTS1), in which the p53 domains that are known to mediate p53 inactivation have been replaced. In this study, the antitumoral activity of CTS1 mediated by adenovirus vector has been evaluated in comparison with a p53 adenovirus vector in various human tumor cell lines. In vitro, in terms of cell growth inhibition, the CTS1 vector was significantly (P < .01) more efficient (2- to 7-fold) than the p53 vector in tumor models overexpressing an inhibitor of p53, murine double minute-2. This result was confirmed in vivo in a pre established tumor developed in nude mice. In an osteosarcoma model overexpressing murine double minute-2, we have shown a significantly (P < .05) higher tumor growth delay with the CTS1 vector compared with the p53 vector (25.6 days compared with 12.4 days). Furthermore, both in vitro and in vivo, we have shown that this higher inhibition of tumor growth with the CTS1 vector was correlated with a higher induction of apoptosis. Therefore, CTS1 is a potentially improved tumor suppressor gene for the treatment of human tumors resistant to wild-type p53 gene therapy. PMID- 10830727 TI - Antitumoral effects of defective herpes simplex virus-mediated transfer of tissue inhibitor of metalloproteinases-2 gene in malignant glioma U87 in vitro: consequences for anti-cancer gene therapy. AB - We set up experiments to evaluate the effects of defective herpes simplex virus (HSV)-mediated in vitro gene transfer of tissue inhibitor of metalloproteinases-2 (TIMP-2) in malignant glioma cells. Intrinsic TIMPs are known to be inhibitors of the strong invasive activities of matrix metalloproteinases in malignant gliomas. The defective HSV vectors dvSRaTIMP2 was engineered to express human TIMP-2 (hTIMP-2) with a combination of replication-competent HSV mutant, temperature sensitive HSV-tsK, and amplicon plasmid-containing hTIMP-2. The hTIMP-2 gene was driven by the simian virus 40 promoter. The helper virus (HSV-tsK) was thermosensitive; consequently, this vector could proliferate only at 31.5 degrees C. After infection of U87 human glioblastoma cells with the vector in vitro, expression of TIMP-2 was confirmed by reverse zymography. The U87 cells infected in vitro either with dvSRaTIMP2 or HSV-tsK were efficiently destroyed under replication-permissive conditions (at 31.5 degrees C) and significantly lowered under replication-nonpermissive conditions (at 37 degrees C). The invasive activity of U87 was clearly inhibited by dvSRaTIMP2 infection at both 31.5 degrees C and 37 degrees C. Our studies suggest that TIMP-2 expressing the defective HSV vector is possibly useful for the treatment of malignant brain tumors. PMID- 10830728 TI - Enhanced green fluorescent protein fusion proteins of herpes simplex virus type 1 thymidine kinase and cytochrome P450 4B1: applications for prodrug-activating gene therapy. AB - To monitor therapeutic transgene expression, we developed fusion genes of enhanced green fluorescent protein (EGFP) with two different prodrug-activating enzyme genes: herpes simplex virus type 1 thymidine kinase (HSV-tk) and rabbit cytochrome P450 4B1 (cyp4b1). Expression of the resulting fusion proteins, TK EGFP and 4B1-EGFP, rendered transduced human and rodent glioma cells sensitive to cytotoxic treatment with the corresponding prodrugs ganciclovir and 4-ipomeanol. Ganciclovir and 4-ipomeanol sensitivity was comparable with that achieved with the native HSV-TK and CYP4B1 proteins. As shown by fluorescence microscopy, TK EGFP was expressed predominantly intranuclearly, whereas 4B1-EGFP was detectable in the cytoplasm, thereby displaying the orthotopic subcellular distribution of the corresponding native enzymes. The fluorescence intensity correlated well with the corresponding prodrug sensitivity, as shown by fluorescence-activated cell sorter analysis. EGFP expression was also used for the selection of stably HSV-tk transduced cells by flow cytometric cell sorting. Resulting cell populations showed a homogeneity of fluorescence intensity similar to single-cell clones after antibiotic selection. In conclusion, tk-egfp and 4b1-egfp fusion genes are valuable tools for monitoring prodrug-activating gene therapy in living cells. EGFP fusion genes/proteins provide a simple and reproducible means for the detection, selection, and characterization of cells expressing enzyme genes for prodrug activation. PMID- 10830729 TI - Stem cell factor and chronic myeloid leukemia CD34+ cells. AB - Normal hematopoiesis is a tightly regulated process involving a balance between signals that stimulate and those that inhibit the proliferation and differentiation of hematopoietic progenitors. In chronic myeloid leukemia (CML) there is a perturbation of these controlling elements, resulting in overgrowth of leukemic cells in the bone marrow and spleen. In part, the proliferation of CML CD34+ cells may result from an abnormal response to the cytokine Stem Cell Factor (SCF). SCF induced proliferation and adhesion to the extracellular matrix via fibronectin are not coupled in CML as they are in normal cells and this may contribute to the accumulation of leukemic progenitors. We have previously shown that CD34+ CML cells and the more primitive CD34+ CD38- CML cells do not require the addition of synergistic cytokines to cultures, but are capable of proliferation in SCF alone, and that leukemic CFU-GM are selectively supported in these cultures. In the presence of other cytokines the response of CML cells to SCF is no greater than that of cells from normal donors, suggesting that the leukemic cells are not more sensitive to SCF, but that accessory pathways are already activated in these cells. Cells from patients with myeloproliferative disorders show variable proliferative response to SCF as the sole mitogenic stimulus, suggesting that expression of bcr-abl is essential for proliferation in this cytokine. Further studies to identify the key determinants of the abnormal response to SCF in CML may lead to a better understanding of the proliferative abnormality that underlies CML. PMID- 10830730 TI - Th2 and Tc2 cells in the regulation of GVHD, GVL, and graft rejection: considerations for the allogeneic transplantation therapy of leukemia and lymphoma. AB - Allogeneic stem cell transplantation (SCT) represents a curative treatment option for patients with leukemia and lymphoma. T lymphocytes contained in the allograft mediate a graft-versus-leukemia (GVL) effect and prevent graft rejection; however, T cells also initiate graft-versus-host disease (GVHD). Identification of T cell populations which mediate a GVL effect and prevent rejection with reduced GVHD will likely improve transplantation outcome. T cells exist in four functionally-defined populations, the CD4+, Th1/Th2 and CD8+, Tc1/Tc2 subsets. Th1-type CD4 cells primarily secrete type I cytokines (IL-2 and IFN-gamma), whereas Th2 cells secrete type II cytokines (IL-4, IL-5, and IL-10). Similarly, the CD8+ Tc1 and Tc2 cells differentially secrete the type I and type II cytokines, respectively. In addition to cytokine secretion, Tc1 and Tc2 populations mediate cytolytic effects, with Tc1 cells utilizing both perforin- and fas-based killing pathways, whereas Tc2 cells primarily utilize perforin mediated cytolysis. In murine transplantation models of graft rejection, GVHD, and GVL effects, we have evaluated such functional T cell subsets for their ability to differentially mediate and regulate transplantation responses. These studies demonstrate that donor Th2 cells do not initiate acute GVHD, and can regulate the GVHD mediated by unmanipulated donor T cells without impairing alloengraftment. Additional experiments have shown that allospecific donor Tc2 cells result in reduced GVHD, and mediate a significant GVL effect. Thirdly, we have demonstrated that non-host reactive Tc2 cells with veto-like activity can potently abrogate marrow rejection independent of GVHD. Together, these results demonstrate that functionally-defined donor Th2 and Tc2 populations play an important role in the regulation of GVHD, the prevention of graft rejection, and the mediation of GVL effects, and suggest that utilization of Th2 and Tc2 cells in clinical allogeneic SCT may have potential for improving treatment outcome. PMID- 10830731 TI - Cell cycle and transcriptional control of human myeloid leukemic cells by transforming growth factor beta. AB - TGFbeta1 is a potent growth inhibitor of both primitive and more differentiated human myeloid leukemic cells. The extent of the growth inhibitory response to TGFbeta varies with cell type, and is not linked to stages of differentiation of cell lines. Downregulation of multiple cell cycle-regulatory molecules is a dominant event in TGFbeta1-mediated growth inhibition of human MV4-11 myeloid leukemia cells. Both G1-phase and G2-phase cyclins and cdks participate in the regulation of TGFbeta1-mediated growth inhibition of MV4-11 cells. By both depressing cdk2 synthesis and up-regulating cyclin E-associated p27, TGFbeta1 may magnify its inhibitory efficiency. TGFbeta1 also rapidly inhibits phosphorylation of pRb at several serine and threonine residues. The underphosphorylated pRb associates with E2F-4 in G1 phase, whereas the phosphorylated pRb mainly binds to E2F-1 and E2F-3 in proliferating MV4-11 cells. Since TGFbeta1 upregulates p130/E2F-4 complex formation and downregulates p107/E2F-4 complex formation, with E2F-4 levels remaining constant, our results suggest that E2F-4 is switched from p107 to pRb and p130 when cells exit from the cell cycle and arrest in G1 by TGFbeta1. In summary, TGFbeta1 inhibits growth of human myeloid leukemic cells through multiple pathways, whereas the "cdk inhibitor" p27 is both a positive and negative regulator. PMID- 10830732 TI - From the study of tumor cell immunogenicity to the generation of antitumor cytotoxic cells in non-Hodgkin's lymphomas. AB - The question of the immunogenicity of non-Hodgkin's lymphoma (NHL) B cells has been investigated in an attempt to support the development of new immunotherapeutic treatments for this disorder, which remains resistant to conventional treatments in most cases. In the present review, we report and discuss our new findings in the field of NHL B cell immunogenicity. One aspect of our work is the description of the expression and functions of membrane molecules associated with antigen presentation. The expression levels of adhesion molecules was measured, and the relevance of this expression to the sensitivity of malignant B cells to cell-mediated lysis was studied. Since the T cell response relies on the expression of both HLA class I and II molecules, we also investigated whether or not these molecules were present at the surface of NHL B cells. Subsequently, we asked whether antitumor CTL and LAK cells could be developed and analyzed the mechanisms of cell lysis involved. Since the generation of a T cell response requires the expression of the costimulatory molecules CD80 and CD86, we investigated their in vivo expression and their modulation in vitro during contact with responding T lymphocytes. The understanding of the immunogenicity of NHL B cells has enabled us to develop a new culture protocol to induce antitumor specific autologous CTL. The originality of NHL B cells--unlike most other tumor cells--is to be able to function as antigen presenting cells (APC) and to activate a T cell response in the absence of other professional APC. Over the next few years, these findings should allow the generation of anti-NHL specific T cells for adoptive immunotherapy and for the identification of NHL-associated antigens. PMID- 10830733 TI - The CD9 molecule on stromal cells. AB - Numerous functions have been attributed to CD9 and other members of the transmembrane 4 (TM4) superfamily. CD9 is thought to be involved in cell proliferation, differentiation, motility and survival. It may also function as part of toxin and virus receptor complexes. Although much remains to be learned about molecular mechanisms, the molecule associates with several integrins, small G proteins, MHC class II molecules and other TM4 superfamily proteins on a given cell surface membrane. Here, we briefly discuss the CD9 displayed on stromal cells that support hematopoiesis and the potential importance of this molecule to osteoclast differentiation. PMID- 10830734 TI - Syndecans and the lymphoid system. AB - Syndecans, transmembrane proteoglycans, play an important role in cell-matrix and cell-cell interactions, as well as modulators in receptor activation. These functions are partly non-specific and related to the heparan sulfate chains attached to the ectodomain, and partly specific due to the transmembrane and cytoplasmic domains of the core protein. In hemopoietic cells syndecan-1 is expressed only in B cells at certain differentiation stages (pre-B and plasma cells). In lymphoproliferative conditions this selective expression is retained in myelomas/plasmacytomas and other lymphoplasmacytic NHL subtypes, and primary effusional lymphomas. It is probably gained in B-CLL, and lost in other NHLs of pre- or post-follicular origin. It is concluded from these empiric results that the expression of syndecan is essential for some NHLs, probably ensuring the required connections to the microenvironment. From a diagnostic point of view, syndecan-1 is a very useful phenotypic marker to indentify cells with plasmacytic differentiation. The importance of syndecan expression in CLL and Hodgkin's lymphoma still requires further studies. PMID- 10830735 TI - Arsenic trioxide therapy for relapsed acute promyelocytic leukemia: an useful salvage therapy. AB - Arsenic trioxide (As2O3) was recently identified as a very potent agent against acute promyelocytic leukemia (APL). Intravenous infusion of 10 mg As2O3 daily for one to two months can induce significant complete remission (CR) of APL, and there is no cross drug-resistance between As2O3 and other antileukemic agents, including all-trans retinoic acid (ATRA). The CR rate of relapsed and/or refractory APL patients who received As2O3 treatment ranged from 52.3% to 93.3%. The median duration to CR ranged from 38 to 51 days, with accumulative As2O3 dosage of 340-430 mg. Although most adverse reactions of As2O3 treatment were tolerable, certain infrequent but severe toxicities related to As2O3 were observed, including renal failure, hepatic damage, cardiac arrhythmia and chronic neuromuscular degeneration, which should be monitored carefully. As2O3 can induce partial differentiation and subsequent apoptosis of APL cells through degradation of wild type PML and PML/RAR alpha chimeric proteins and possible anti mitochondrial effects. Like the treatment of ATRA in APL, early relapses from As2O3 treatment within a few months were not infrequently seen, indicating that rapid emerging resistance to As2O3 can occur. Nevertheless, the PML/RAR alpha fusion protein was reported to disappear in some APL patients who received As2O3, and who might earn long-survival. However, the follow-up is still too short to draw the conclusion. Intriguingly, it has been shown that As2O3 can also induce apoptosis of other non-APL tumor cells with clinical achievable concentrations. However, the detailed molecular mechanisms are not yet fully understood. Further studies regarding to the pharmacological characters, clinical efficacies, toxicities, apoptogenic mechanisms, and spectrum of anti-tumor activity of As2O3 are warranted. PMID- 10830736 TI - Detection of residual disease in pediatric B-cell precursor acute lymphoblastic leukemia by comparative phenotype mapping: method and significance. AB - The present review summarizes our efforts in developing a novel immunologic approach ("Comparative Phenotype Mapping") targeted at assessing minimal residual disease (MRD) in B-cell precursor (BCP) acute lymphoblastic leukemia (ALL) patients. The method relies on quantitatively aberrant, leukemia-associated antigen expression patterns which allow to discriminate leukemic from normal BCP using a limited panel of antibody combinations and multidimensional flow cytometry. In an analysis of 63 follow up bone marrow samples of patients with BCP-ALL we show that this approach enables to efficiently detect MRD. Further clinical observation revealed that the patients which were MRD-positive by flow cytometry (although in morphological remission) had a very high probability of early disease recurrence compared to the good chances of a relapse-free survival (RFS) in the MRD-negative cohort (RFS 0.0 vs. 0.76 at 3 years). Comparative Phenotype Mapping thus proves to be a reliable method for MRD detection in BCP ALL. Concluding remarks relate to the optional applications of the method as well as to future perspectives. An ongoing large prospective study which we are now conducting on the basis of Comparative Phenotype Mapping will clarify the clinical significance of MRD detection in ALL patients by this method, and will determine its value compared to related as well as molecular-genetic techniques. PMID- 10830737 TI - Phase II evaluation of a high-dose mitoxantrone based induction regimen in untreated adults with acute myeloid leukemia. AB - To evaluate a regimen including high-dose mitoxantrone in previously untreated adults with AML, 45 patients aged 21-59 (median 41) were given cytarabine, 3 g/m2 days 1-5, mitoxantrone, 80 mg/m2 day 2 and etoposide, 150 mg/m2 days 1,3,5. Post remission therapy consisted of 5 cycles combining the same agents at reduced doses. Complete remission was seen in 36 patients. The observed 3-year survival is 28%. Cytogenetic pattern and CD34 expression correlated with response and survival. Significant toxicity included myelosuppression, mucositis, diarrhea and hyperbilirubinemia. Ventricular ejection fraction was generally reduced, with clinical cardiac dysfunction in only 2 patients. This high-dose mitoxantrone combination can be administered to young adults with AML with tolerable toxicity and results comparable to those of other dose-intensive regimens. PMID- 10830738 TI - Expression of cytotoxic proteins in peripheral T-cell and natural killer-cell (NK) lymphomas: association with extranodal site, NK or Tgammadelta phenotype, anaplastic morphology and CD30 expression. AB - Most peripheral T-cell lymphomas (PTCL) express the alphabeta T-cell receptor (TCR) whereas rare PTCL express the gammadelta TCR. Most if not all gammadelta PTCL are extranodal lymphomas and among them, hepatosplenic gammadelta PTCL constitute a distinct clinicopathological entity. Besides alphabeta and gammadelta PTCL, there is a recently recognized group of extranodal, mainly nasal tumours, which display, in most instances, phenotypic and genotypic features of Natural-Killer cell non-Hodgkin's lymphomas (NK-NHL). Cytotoxic cells, including NK cells and cytotoxic alphabeta and gammadelta T lymphocytes may induce lysis of the target by using granule-associated cytotoxic proteins such as the T-cell intracellular antigen-1 (TIA-1), perforin and granzyme B. Expression of TIA-1 can be detected in all cytotoxic cells whereas granzyme B and perforin expression can be detected in high levels only in activated cytotoxic cells. Recently, several studies showed that the expression of these cytotoxic proteins in tumour cells of PTCL and NK-NHL is associated with a) extranodal site of clinicopathological presentation b) NK or Tgammadelta-cell phenotype c) CD30 expression in cutaneous T-cell lymphoproliferations and d) anaplastic morphology in nodal PTCL. This latter finding contrasts with the data that only rare Hodgkin lymphomas (HL) express cytotoxic proteins in Hodgkin and Reed-Sternberg cells. Altogether the data of the literature indicate that most extranodal T and NK-NHL are activated cytotoxic lymphomas with the notable exception of hepatosplenic gammadelta PTCL which represent tumours of non-activated cytotoxic cells. On this basis, it is suggested that the expression of cytotoxic proteins may be useful for the identification and classification of extranodal T and NK-cell lymphomas and, to some extent, for the differential diagnosis between HL and CD30+ anaplastic large cell lymphomas. Cytotoxic lymphomas are preferentially localized in extranodal sites such as skin, lung, upper respiratory and gastrointestinal tracts, which are continuously exposed to various antigens. Since cytotoxic T and NK cells are regarded as first line of defense in these sites, and some cytotoxic tumours such as nasal lymphomas and enteropathy-type intestinal lymphomas are associated with EBV and gliadin, respectively, it is likely that chronic antigen exposure may play a role in the pathogenesis of cytotoxic lymphomas occurring in mucosa and/or skin. Besides chronic antigenic stimulation, chronic immunosuppression may also have pathogenetic significance in cytotoxic lymphomas in view of their increased incidence in immunocompromised patients. PMID- 10830739 TI - A comparison of elderly patients with aggressive histology lymphoma who were entered or not entered on to a randomized phase II trial. AB - The purpose of this study was to compare the baseline patient characteristics, treatments and outcomes of elderly patients with aggressive histology lymphoma who were entered or not entered onto a randomized phase II trial. We previously conducted a randomized phase II trial in patients > or = 65 years of age who had advanced stage intermediate grade lymphoma. A registry of all patients meeting the inclusion criteria for that trial was maintained. Many patients were not entered on to the randomized trial because of the presence of at least one exclusion criterion, or because of patient or physician choice. We have compared the baseline characteristics, treatment, and survival of the randomized and non randomized patients. Results show that 68 consecutive patients met inclusion criteria for the randomized trial. Thirty-eight patients satisfied all eligibility criteria, consented, and were randomized; 30 patients (44%) were not entered. In comparison with randomized patients, non-randomized patients were older (mean 75.9 vs. 72.4 years; P=0.013), had a poorer performance status (P=0.0006), were less likely to be given treatment with curative intent (60% vs. 100%; P<0.001), and were less likely to complete 6 cycles of such treatment (27% vs. 89%; P<0.001). With a median follow-up of > 7 years, actuarial 5-year survival is superior in randomized patients (44.3% vs. 10%; P<0.00001). In conclusion, a substantial number of patients did not enter our randomized trial phase II trial and had different characteristics, received different therapy and had inferior outcomes in comparison with randomized patients. Randomized trials of therapy for elderly lymphoma patients may include special selection criteria and results may not be generalizable to a substantial proportion of other older patients. PMID- 10830740 TI - Predictive factors for central nervous system involvement in non-Hodgkin's lymphoma: significance of very high serum LDH concentrations. AB - Factors predictive for central nervous system (CNS) involvement at presentation were investigated in 152 patients with non-Hodgkin's lymphoma (NHL) except for lymphoblastic cell lymphoma and small noncleaved cell lymphoma. Twelve patients developed CNS involvement during their disease course. The incidence was 7.9% of all the patients studied and 17.0% of the patients with serum LDH concentration > or = two times the upper limit of normal (2N). By univariate analysis, stage IV disease (P = .023), a serum LDH concentration > or = 2 N (P = .009), and bone marrow involvement (P = .016) were risk factors for CNS involvement. Multivariate logistic regression analysis identified a serum LDH concentration > or = 2 N (P = .032) as an independent predictor for CNS involvement. All 12 patients who developed CNS involvement were among the 126 patients with diffuse lymphoma, whereas none of the 17 patients with follicular lymphoma developed CNS involvement, although the difference was not statistically significant. The median survival of the patients with CNS involvement was only 4.5 months. We conclude that a serum LDH concentration > or = 2N at presentation is a significant predictive factor for CNS involvement for NHL patients without lymphoblastic lymphoma and small noncleaved cell lymphoma. Therefore, we would suggest that CNS prophylaxis should be considered for patients with a serum LDH concentration > or = 2N at presentation and diffuse lymphoma once a complete remission is achieved. PMID- 10830741 TI - A prospective study of CD38/45 flow cytometry and immunofluorescence microscopy to detect blood plasma cells in patients with plasma cell proliferative disorders. AB - Malignant plasma cells can be detected in the blood of patients with multiple myeloma (MM) using flow cytometry (FC), immunofluorescence microscopy (IM), or a variety of molecular techniques. Increased numbers of light chain-restricted blood plasma cells as detected by IM is associated with a diagnosis of overt MM and a decreased overall survival. The IM technique is time consuming; therefore, a prospective study was designed to test whether CD38 CD45 FC could simplify the procedure. Blood samples from 769 patients with plasma cell proliferative disorders were studied prospectively by FC and IM over a one-year period. The FC technique was performed on 1 ml of whole blood after ammonium chloride red blood cell lysis and utilized anti-CD38PE and anti-CD45PerCP. The number of CD38+ 45- events were enumerated and compared to the number of light chain-restricted plasma cells detected by the standard IM technique. In 46% (353/769) of cases > or = 1 CD38+ CD45- events were detected by FC whereas IM was positive for light chain restricted plasma cells in 33%; there was concordance between FC and IM in 73% of cases. In 20% of cases FC was positive and IM was negative; however, in 7% of cases FC was negative yet light chain-restricted plasma cells were detected by IM. FC was positive in 88% (134/153) of cases where the IM technique showed a high number of circulating plasma cells. This study demonstrates that two-color CD38/45 FC identifies most cases with a high IM result and reduces the workload in the clinical laboratory. The prognostic implications of a positive FC screen but a negative IM will require long-term patient follow-up. PMID- 10830742 TI - Morphologic, immunophenotypic and in vitro growth characteristics of blood and bone marrow associated with stem cell mobilisation in patients with lymphoma. AB - The proportion of CD34+ cells in the bone marrow (BM) is predictive of the size of progenitor cell mobilisation into the blood (PB). To investigate which other PB and BM parameters may be related to mobilisation, we analysed at steady state PB and BM of 23 patients with relapsed or resistant lymphoma before administering high-dose cyclophosphamide and G-CSF Cell morphology, number of CD34+ cells, and growth in clonogenic assay and in long-term cultures (LTC) were determined and then correlated with mobilisation extent (CD34+ and GM-CFC) and quality (growth of harvested cells in LTC). We found that the good mobilising patients (CD34 > 50 x 10(3)/ml, n=10) had several baseline BM characteristics (number of CD34+ MNC, GM-CFC, BFU-E, production of CFCs in LTC) similar to a group of 12 healthy controls, while patients with reduced mobilisation (CD34 < 50 x 10(3)/ml, n=13) had clearly reduced BM progenitors and LTC growth (p< 0.05). In a multivariate analysis including baseline clinical, blood and bone marrow characteristics, the most significant PB and BM factors independently associated with a higher number and/or quality of mobilised cells were a higher number of CD34+ and GM-CFC in the BM and a higher baseline haemoglobin, platelet, and CD34+ blood count. The capacity to release progenitor cells into the circulation is therefore not predicted by the distribution of morphologically distinguishable cells, marginally predicted by the BM content of highly undifferentiated cells (growth in long term culture), while it is proportional to the number of BM progenitors (CD34+, GM-CFC and BFU-E). PMID- 10830744 TI - Molecular analysis of latent membrane protein 1 in patients with Epstein-Barr virus-associated hemophagocytic lymphohistiocytosis in Japan. AB - Latent membrane protein 1 (LMP1) of Epstein-Barr virus (EBV) is considered to be an oncoprotein because it is crucial for B-lymphocyte transformation. Since a 30 base pair (bp) deletion in the carboxy-terminal portion of the LMP1 gene was found in a CAO cell line derived from nasopharyngeal carcinoma containing EBV, an association between EB viral genetic alteration and tumorigenicity has been postulated. In this study we have analyzed LMP1 DNA isolated from 10 Japanese patients with EBV-associated hemophagocytic lymphohistiocytosis (EBV-HLH). In all HLH patients, we found the 30 bp deletion and 4-8-tandem repeats of the sequence DNGPQDPDNTD in the LMP1 gene. Furthermore, detailed amino acid (aa) sequence analysis revealed that 7 aa substitutions identical to those found in CAO-LMP1 but not in B95.8 cell line-LMP1 were found in all the HLH cases. NF-kappaB assay revealed that HLH-LMP1 activated NF-kappaB significantly more than that of B95.8 LMP1 (p=0.032). We conclude that EBV from all of our HLH cases shared common genetic characteristics with EBV obtained from the CAO cell line, which is distinct from the wild-type EBV isolated from the B95.8 cell line. These data suggest that the mutational changes of the LMP1 gene may play an important role in the pathogenesis of these fatal EBV-related disorders. PMID- 10830743 TI - The role of chemokines in Hodgkin's disease. AB - Recent studies have analyzed the expression of chemokines in tissues involved by Hodgkin's disease (HD) (1). The data indicate a significant role for chemokine expression in the pathobiology and pathophysiology of HD. In general, HD tissues showed higher levels of chemokine expression than reactive lymphoid hyperplasia (RLH) tissues. There were major differences in chemokine expression among the different HD subtypes. Similar to previous studies in athymic mice that identified a pattern of chemokine response induced by Epstein-Barr virus (EBV) infected cells, the expression of IP-10, Mig, RANTES, and MIP1-alpha was higher in EBV positive compared to EBV negative HD tissues. In addition, there was a direct correlation of eotaxin expression with tissue eosinophilia. By immunohistochemistry, IP-10 and Mig proteins localized in the malignant Reed Steinberg (RS) cells and their variants, and to some surrounding inflammatory cells. Eotaxin localized to fibroblasts and smooth muscle of blood vessels. In this review, we discuss the patterns of expression of IP-10, Mig, RANTES, MIP1 alpha, and eotaxin in HD and its subtypes, and the relationship to EBV positivity, LMP1 expression, tissue eosinophilia and T cell infiltration. In addition, we discuss the potential role of chemokines and cytokines in the pathobiology of HD. PMID- 10830745 TI - Human T-lymphotropic virus type I provirus and T-cell prolymphocytic leukemia. AB - T-cell prolymphocytic leukemia (T-PLL) is a rare type of post-thymic T-cell neoplasm, the etiology of which is unknown. Patients with T-PLL have been found to be seronegative for human T-lymphotropic virus type-I (HTLV-I), and their leukemia cells do not retain monoclonally integrated HTLV-I provirus. Recently, we have demonstrated the presence of defective HTLV-I provirus by polymerase chain reaction in the DNA extracted from peripheral blood cells or affected lymph nodes of T-PLL patients. Although there is a possibility, from our observation, that an alternative mechanism is operating in HTLV-associated leukemogenesis, it is still unknown whether and how HTLV-I can contribute to the leukemogenesis of T PLL. In this review, we describe controversial issues and discuss a role of HTLV I in the leukemogenesis of T-PLL. PMID- 10830746 TI - Increased expression of vascular endothelial growth factor (VEGF) in Castleman's disease: proposed pathomechanism of vascular proliferation in the affected lymph node. AB - Castleman's disease is a lymphoproliferative disorder of unknown etiology characterized by enlarged hyperplastic lymph nodes with marked vascular proliferation. To evaluate the possible involvement of vascular endothelial growth factor (VEGF) in the pathogenesis of Castleman's disease, we studied VEGF expression in sera and lymph nodes from four patients with either the plasma-cell type or mixed type of Castleman's disease. Clinically, one patient had the multicentric type and the others the localized type. The VEGF levels of the sera and the supernatants of the cultured lymph nodes were higher than those of normal controls. VEGF was strongly expressed in plasma cells in the interfollicular region of the lymph nodes, but rarely in normal lymph nodes. The disregulated IL 6 gene expression is considered to be a primary event that could be related to the etiology of this disease. Recently, Kaposi's sarcoma virus/human herpes virus 8 (KSHV/HHV-8) has been reported to be associated with a subset of the multicentric type of Castleman's disease, and a viral homologue of IL-6 (vIL-6) encoded by KSHV/HHV-8 has been shown to induce VEGF expression. Human IL-6 produced in the affected lymph nodes of Castleman's disease may induce paracrine VEGF-production by plasma cells and vascular proliferation in the lymph node. The confirmation of the role of VEGF in the pathogenesis of Castleman's disease may provide a therapeutic strategy. PMID- 10830747 TI - Normal structure of NFKB2, C-REL and BCL-3 gene loci in lymphoproliferative and myeloproliferative disorders. AB - NF-kappaB/rel transcription factors are crucial regulators of development, differentiation and apoptosis of both lymphoid and myeloid lineages. There is increasing evidence for an involvement of NF-kappaB/rel proteins in lymphomagenesis and resistance of lymphoid tumors to the induction of apoptosis. Structural alterations of the NF-kappaB/rel genes NFkappaB2, c-rel and bcl-3 have been shown to result in increased NF-kappaB/rel activity. Because we observed strong constitutive NF-kappaB/rel binding activity in chronic lymphocytic leukemia of the B-cell type (B-CLL) which may contribute to resistance against cytotoxic drugs we studied the genomic organisation of NFkappaB2, c-rel and bcl-3 gene loci in a panel of lymphoproliferative disorders (n=81) with an emphasis on B-CLL (n=47). The method of genomic Southern blotting using cDNAs of the respective genes was used. In spite of the role of NF-kappaB/rel in myeloid maturation there is no data available as to the occurrence of NF-kappaB/rel rearrangements in chronic myeloproliferative syndromes (cMPS). For this reason we included a small panel of cMPS patients (n=16). Southern Blotting revealed a germline configuration of NFkappaB2, c-rel and bcl-3 loci in all NHL and cMPS patients examined. Our results demonstrate that structural alterations of NFkappaB2, c-rel and bcl-3 genes at the Southern Blotting level are rare events that do not contribute to lymphoid or myeloid transformation in the majority of NHL or cMPS patients. PMID- 10830749 TI - Lack of integrated TT virus (TTV) genomes in cellular DNA in infected human hematopoietic cells. AB - TT virus (TTV) isolated from the serum of a patient with posttransfusion hepatitis has been characterized as a member of the Circoviridae, a family of small DNA viruses with single-stranded circular genomes. TTV appeared to infect not only the serum and liver, but also the peripheral blood mononuclear cells (PBMC). We investigated the prevalence of TTV DNA in human hematopoietic cells, based on 84 mononuclear cell samples obtained from the bone marrow or lymph nodes of patients with hematopoietic malignancies including leukemia, malignant lymphoma and aplastic anemia. Forty-nine (58.3%) out of the 84 samples were positive for TTV DNA with polymerase chain reaction analysis, which was almost similar to the frequency found in the patients' serum. Southern blot analyses using a 3.2-kb fragment derived from the TTV DNA, however, showed no evidence supporting the fact that the TTV genomes are integrated into the human hematopoietic cell genomes, thus suggesting their existence as episomal forms. PMID- 10830748 TI - Virological and molecular characterisation of a new B lymphoid cell line, established from an AIDS patient with primary effusion lymphoma, harbouring both KSHV/HHV8 and EBV viruses. AB - We report here a new case of primary effusion lymphoma (PEL), occurring in a French homosexual HIV-1 infected male with a pericardial, pleural and mesenteric tumour dissemination, and the establishment from his pleural effusion of a new cell line, Cra-BCBL, dually infected by EBV and KSHV/HHV8. Cra-BCBL cells are of B-cell origin as judged by their clonal immunoglobulin heavy chain (IgH) gene rearrangement, identical to that of the parental tumour. Both the cell line and the lymphoma cells expressed CD38 and CD45 antigens but no classical B-cell or T cell lineage-restricted antigens. Cra-BCBL harbours a type I EBV virus, expressing a latency type II. Expression of KSHV/HHV8 ORF72 and ORF75 was detected by RT/PCR. In addition, KSHV lytic replication could be induced by treatment by n-butyrate. An equivalent and high copy number of KSHV genomes (20 to 200 copies by cell) was detected both in the primary tumour cells and in the cell line. Southern blot (SB) analysis of EBV terminal repeats (TR) displayed the same unique band in the cell line DNA and in the original tumour cells, consistent with a monoclonal infection of EBV. Furthermore, SB analysis of KSHV/HHV8 TR revealed the same hybridisation pattern between Cra-BCBL and the effusion cells, with a common band at around 30-40 kb corresponding to the fused termini of the viral episomes and a 5 Kb rearranged fragment. The new cell line characterised here could be a useful model to study interactions between two human herpes viruses and their contribution to lymphomagenesis. PMID- 10830750 TI - Non-Hodgkin's lymphoma presenting as anasarca: probably mediated by tumor necrosis factor alpha (TNF-alpha). AB - Two patients presented with anasarca, fevers and sweats. Subsequent evaluation revealed aggressive lymphoproliferative disease. Both patients were treated with CHOP chemotherapy. One patient responded with spontaneous, vigorous diuresis and complete resolution of the edema. She relapsed two months later with recurrent edema that responded a second time to salvage chemotherapy. The second patient died of gram positive sepsis a week after diagnosis. As anasarca is an unusual presenting symptom of non-Hodgkin's lymphoma, we postulated that the malignant cells were secreting a cytokine that resulted in "vascular leakage" of fluid and development of diffuse edema. Several serum cytokine levels were tested. Both patients had elevated TNF-alpha levels, which could have been the cause of the edema; or there might be yet another unidentified mediator that was responsible for the anasarca. We report these two cases to bring to attention the unusual nature of this presentation. PMID- 10830751 TI - Immunoglobulin related amyloidosis presenting as isolated lymph node and pulmonary involvement. AB - Here we present an unusual case of a 53-year old patient presenting AL-kappa amyloidosis with diffuse-type amyloidosis of lungs, lymph nodes and pleura. The underlying pathology was a B-cell immunoglobulin-secreting non-Hodgkin lymphoma, as proven by the presence of a monoclonal B-cell population in the bone marrow. Diffuse parenchymal infiltration of the lungs is extremely rare in non-systemic amyloidosis, with only 4 previous cases having been reported in the English literature. PMID- 10830752 TI - Treatment of deep vein thrombosis using temporary vena caval filters after allogeneic bone marrow transplantation. AB - Bone marrow transplant (BMT) recipients have risk factors for deep vein thrombosis (DVT) including venous stasis caused by immobilization in the sterile unit, vessel wall damage caused by preparative regimen or indwelling catheters, and hypercoagulability caused by decreased natural anticoagulants. We successfully treated a patient who developed massive DVT in the superior vena cava after BMT with anticoagulation and the use of temporary vena caval filters. Considering the delayed complications, permanent filter is not appropriate for BMT recipients, because the risk factors for DVT associated with BMT are transient. We considered that temporary vena caval filter is a safe and useful device to prevent pulmonary embolism after DVT in BMT recipients. PMID- 10830753 TI - The effect of docarpamine, a dopamine pro-drug, on blood pressure and catecholamine levels in spontaneously hypertensive rats. AB - We studied the effects of bolus intravenous injection of the dopamine prodrug, docarpamine (200 microg/kg), on mean arterial pressure (MAP) and heart rate (HR) in Wistar-Kyoto (WKY) and spontaneously hypertensive rats (SHRs). In WKY rats (n=18), MAP and HR increased 5 min after docarpamine and then returned to baseline levels within 15 min. In contrast, in SHRs (n=15), MAP and HR gradually decreased, reaching a nadir 20 min after injection. Five min after docarpamine, plasma dopamine and 3,4-dihydroxy phenyl acetic (DOPAC) levels increased in both WKY rats (n=5) and SHRs (n=5). The docarpamine-induced changes in MAP and HR in both rat strains (n=5/strain) were blocked by the D1-like antagonist, SCH23390. alpha-Adrenergic (n=4) and vasopressin V1 (n=3) receptor blockade also abrogated the effects of docarpamine in WKY rats. We conclude that docarpamine differentially affects MAP and HR in WKY and SHRs. In SHRs, the depressor and bradycardiac effects of docarpamine are mediated by D1-like receptors. In WKY rats, the pressor and tachycardiac responses are caused by an interaction among D1-like, alpha-adrenergic, and V1 receptors. PMID- 10830754 TI - Post-exercise elevations in sympathetic nerve activity and baroreflex function in normotensive rabbits. AB - We tested the hypothesis that a single bout of dynamic exercise reduces post exercise arterial pressure, heart rate, and renal sympathetic nerve activity and attenuates the arterial baroreflex control of heart rate and renal sympathetic nerve activity in normotensive New Zealand White rabbits. Animals were chronically instrumented with right jugular venous and left femoral arterial catheters, and electrodes around the renal sympathetic nerve. Arterial pressure, heart rate, and renal sympathetic nerve activity were recorded for two hours pre exercise and two hours after a single bout of treadmill exercise (post-exercise). Post-exercise heart rate, arterial pressure, and renal sympathetic nerve activity were elevated above pre-exercise values (71+/-3 bpm, 13+/-1 mmHg, and 80+/-21%, respectively). These data demonstrate that normotensive rabbits do not exhibit post-exercise hypotension, due in part to elevations in sympathetic nerve activity. In addition, arterial baroreflex regulation of heart rate and renal sympathetic nerve activity were determined pre- and post-exercise. Exercise shifted the baroreflex function curve for heart rate and renal sympathetic nerve activity upward and to the right without a change in gain. These data suggest that post-exercise elevations in sympathetic nerve activity are due, in part, to an elevation of the operating point of the arterial baroreflex to a higher pressure. These responses in normotensive rabbits contrast sharply with the responses in hypertensive individuals and animals. Understanding the mechanisms contributing to the differences between hypertensive and normotensive subjects may lead to measures designed to lower arterial pressure in hypertensive individuals. PMID- 10830755 TI - Epidemiological risk factors of coronary heart disease are not causal in atherosclerosis. AB - Logic dictates that for scientific progress in atherogenesis "cause" must be the sole prequisite without which the disease cannot occur. Nor can it be assumed that statistical associations (risk factors) with coronary heart disease (CHD) are causal for atherosclerosis and extrapolations from correlations with CHD incidence to atherosclerosis are invalid. Any factor considered to play a role in atherogenesis requires pathological and experimental evidence consistent with the logic of Koch's specificity of cause and effect. Current epidemiological misuse and manipulation of cause and risk factors are contrary to the basic precepts of scientific logic and the fundamental need for precision in word usage. The term "risk factor", because of the current deeply entrenched false concept of causality has retarded medical progress and should be abandoned. Its adherents, guilty of a disservice to the tenets of their discipline, have also sullied the scientific integrity of medicine as a whole. PMID- 10830756 TI - To use Internet in collaborative studies and registers. PMID- 10830757 TI - A prospective randomized study comparing acupuncture with physiotherapy for low back and pelvic pain in pregnancy. AB - BACKGROUND: The aim of this study was to describe the effects of acupuncture in the treatment of low-back and pelvic pain during pregnancy and compare it with physiotherapy. METHODS: Sixty pregnant women were allotted to acupuncture or physiotherapy. The women estimated the severity of their pain using a visual analog scale (VAS) from 0 to 10 and disability in performing twelve common daily activities using a disability-rating index (DRI) from 0 to 10. RESULTS: In the acupuncture group all 30 women completed the study (two exclusions), in the physiotherapy group only 18. Before treatment the two study groups were rather similar with respect to pain and disability. After treatment the mean morning VAS had declined from 3.4 to 0.9 (p<0.01) in the acupuncture group and from 3.7 to 2.3 (NS) in the physiotherapy group. The corresponding evening values had declined from 7.4 to 1.7 (p<0.01) and 6.6 to 4.5 (p<0.01), respectively. The mean VAS values were lower after acupuncture than after physiotherapy both in the morning (p=0.02) and in the evening (p<0.01). After treatment also the mean DRI values had decreased significantly in the acupuncture group for 11 of 12 activities and the values were significantly lower for all activities than in the physiotherapy group where no significant changes had taken place. Overall satisfaction was good in both groups. There were no serious adverse events in any of the patients. CONCLUSIONS: Acupuncture relieved pain and diminished disability in low-back pain during pregnancy better than physiotherapy. PMID- 10830758 TI - Fetal oxygen saturation during epidural and paracervical analgesia. AB - BACKGROUND: We wanted to assess changes in fetal oxygenation during maternal epidural or paracervical analgesia in labor. METHODS: A prospective, open and non randomized study. Twenty healthy parturients were enrolled before they asked for pain relief. Informed consent was obtained. Fetal and maternal oxygen saturations were measured before and up to 1 h after the initiation of analgesia. Fetal oximetry was performed with the Nellcor N-400 oximeter+FS-14B fetal oxygen sensor (Nellcor Puritan Bennett, Pleasanton, California, USA). Maternal oximetry was done with Datex Satlite portable monitor (Datex, Finland). Visual analog scale was used for assessing pain relief. Two-way analysis of variance and students t test were used for statistical analyses. RESULTS: Fetal oxygenation initially improved in both groups. The saturation then returned to baseline in both groups. In the epidural group, the values remained at baseline or slightly below, while in the paracervical group the saturation remained a little higher than baseline (p=0.009). No change was seen in maternal oxygenation or heart rate. No change in fetal heart rate was found either. Epidural block was superior to paracervical block with respect to pain relief (p=0.002). CONCLUSIONS: There was a small but significant difference in fetal oxygenation between epidural and paracervical groups during the observation period. The magnitude of the difference is hardly clinically significant. A larger, randomized study is needed to elucidate the mechanisms behind this finding. PMID- 10830759 TI - The prevalence of anemia in pregnant Nepali women--a study in Kathmandu. AB - BACKGROUND: Anemia in pregnancy is associated with maternal and perinatal morbidity and mortality. The World Health Organization has suggested that where up-to-date information is not available, prevalence studies should be undertaken. METHODS: The distribution of hematocrit in 2280 pregnant women attending Patan Hospital, Kathmandu, for their first antenatal visit in a twelve month period, 1994-95, was studied. Anemia was defined as a hematocrit value < or =34% in the first and third trimesters, or < or =33% in the second trimester. Severe anemia was defined as a hematocrit value < or =24%. The prevalence of anemia and severe anemia, and associations with socio-demographic (age, ethnic group, living area, maternal and paternal education and occupation) and biological (height, body mass index, parity, gestation) variables were investigated. RESULTS: The hematocrit values ranged from 11-49% with a mean value of 32.6% (s.d. 3.9). The prevalences of anemia and severe anemia were 62.2% and 3.6%, respectively. High prevalence of anemia was observed among teenagers, farmers, women of short height, the ethnic groups Lama/Sherpa/Tamang, and women married to industrial workers or illiterate men. Also, the risk of anemia increased with gestation. Work within the service professions, higher education and high body mass index, were associated with a lower risk of anemia. CONCLUSIONS: A high prevalence of anemia among pregnant women in Kathmandu was observed. The hematocrit values were related to socio demographic and biological factors. In order to plan focused and effective intervention, studies on the etiology of anemia among pregnant women in rural and urban areas of Nepal need to be carried out. PMID- 10830760 TI - Relationship between histopathological changes in post partum renal biopsies and renal function tests of African women with early onset pre-eclampsia. AB - BACKGROUND: To improve the diagnostic accuracy of concurrent renal disease in hypertension of pregnancy, biopsy evaluation is essential. In addition, establishing underlying renal disease is important for prognosis on future pregnancies. We therefore designed a study to determine the diagnostic yield of postpartum renal biopsy and the nature and frequency of complications associated with this procedure. Also, to determine relationships, if any, between renal function tests and ultrastructural and histopathological findings. METHODS: Fifty renal biopsies were performed in the immediate postpartum period in black African women with early onset pre-eclampsia. Each biopsy specimen was placed in a separate container and coded so that sampling was unknown to the electron microscopist. Each biopsy specimen was divided into three parts, and processed and stained for light, fluorescent and transmission electron microscopy using conventional techniques. RESULTS: Renal tissue biopsies were adequate for diagnostic purposes in all cases. There were no complications in any of the 50 patients studied. Ultrastructural examination confirmed the light microscopy findings. In addition the ultrastructural findings showed intramembranous deposits, foot process fusion and mesangial deposits. In 16 patients with normal renal function tests; the biopsies evaluation from these patients showed ultrastructural changes. In the remaining 34 patients with abnormal renal function tests of varying severity; biopsy evaluation from these patients showed both ultrastructural and histopathological changes. CONCLUSION: Renal biopsy procedure is safe, and ultrastructural and histological findings obtained from postpartum renal biopsies are more informative than the routine renal function tests. PMID- 10830761 TI - Effect of lactoferrin on lipopolysaccharide (LPS) induced preterm delivery in mice. AB - BACKGROUND: The present study attempted to confirm the preventive effect of lactoferrin on lipopolysaccharide (LPS) induced preterm delivery in mice. METHODS: Female C3H/HeNCrj mice were pair-mated with male Crj: B6D2F1 mice. On day 15 of gestation, a 50 micron/kg intraperitoneal injection of LPS (LPS group) was administered twice with a 3-hour interval between injections (at 2:00 and 5:00 PM). The mice were also given an intraperitoneal injection of lactoferrin (0.2 or 1 mg/200 microl/body) 1 hour prior to each LPS injection) (at 1:00 and 4:00 PM). To determine the effect of LF on plasma IL-6 level, mother mice were sacrificed and blood samples were obtained at 6 hours after the second treatment of LPS. RESULTS: Preterm delivery was induced by LPS in all mice on day 16 of gestation. Lactoferrin administration to LPS-treated mice significantly prolonged (p<0.001) gestation when compared with the LPS group. Plasma levels of IL-6 in the LPS group (1.628+/-115 pg/ml) were significantly higher (p<0.001) than in the untreated group (497+/-39 pg/ml). On the other hand, administration of lactoferrin (1 mg/body) to LPS-treated mice significantly suppressed (p<0.001) IL 6 levels (1.060+/-154 pg/ml). CONCLUSIONS: Lactoferrin exerts a preventive effect on the incidence of preterm delivery in mice via a suppression of plasma IL-6 augmentation by LPS. PMID- 10830762 TI - Mean cell volume and gamma-glutamyl transferase are superior to carbohydrate deficient transferrin and hemoglobin-acetaldehyde adducts in the follow-up of pregnant women with alcohol abuse. AB - BACKGROUND AND OBJECTIVE: To compare the usefulness of carbohydrate-deficient transferrin (CDT), the ratio of CDT to total transferrin, and hemoglobin acetaldehyde adducts with mean cell volume (MCV) and gamma-glutamyl transferase (GGT) in the follow-up of alcohol abuse during pregnancy. METHODS: Forty-four pregnant drug and alcohol abusing female patients attending a special outpatient clinic were followed from the 8th to 24th gestational week onwards. A population of sixty-two healthy pregnant women was recruited to assess the effect of gestation on the markers. RESULTS: Eight of thirteen heavy drinking (> or =8 drinks/week) patients delivered infants with fetal alcohol effects (FAE). MCV and GGT were higher among heavy drinking patients than in moderately drinking (<8 drinks/week) patients (92+/-4 vs 90+/-3 fl and 31+/-34 vs 16+/-10 U/ L, respectively), and in patients delivering infants with FAE compared with patients delivering healthy infants (95+/-3 vs 90+/-3 fl and 34+/-26 vs 15+/-10 U/L, respectively). Hemoglobin-acetaldehyde adducts, CDT, and the ratio of CDT to total transferrin were neither associated with the reported level of alcohol consumption nor with the occurrence of FAE. In the receiver operating characteristics analysis MCV was found to be superior to CDT and the adducts, and GGT superior to the adducts, in identifying heavy drinking and in predicting FAE. In the control population, both CDT and total transferrin were found to rise during pregnancy, whereas the ratio of CDT to total transferrin was found to decline. The upper reference range of 33 U/L for CDT was considerably higher than that of non-pregnant women (26 U/L). CONCLUSION: MCV and GGT appear to be the most efficient laboratory markers for detecting excessive alcohol consumption and the adverse effects of alcohol on the fetus. PMID- 10830763 TI - Diabetic retinopathy in pregnancy during tight metabolic control. AB - BACKGROUND: The relation between retinopathy and the parameters: 24-h blood pressure, glucose control, albuminuria, and outcome of pregnancy was studied before, during, and after pregnancy in women with insulin-dependent diabetes mellitus on tight metabolic control during pregnancy. METHODS: Prospective study of 112 pregnant women with insulin-dependent diabetes mellitus followed with fundus photography at the Department of Ophthalmology, Arhus University Hospital. Changes in retinopathy were related to 24-h blood pressure, blood glucose, albuminuria, and adverse perinatal outcome. RESULTS: There was an association between grade of retinopathy and HbAlc before (Spearman's rho=0.49, p<0.04) and after pregnancy (Spearman's rho=0.42, p<0.02), but no such correlation was found at any examination during pregnancy where glycemia was kept tight. Those women who had progression of retinopathy during or after pregnancy had significantly earlier onset of diabetes mellitus (14+/-8 years, range 1-27) than those women with improvement or no progression of retinopathy (19+/-8 years, range 1-36, p<0.04). No association was found between progression of retinopathy and HbA1c, blood pressure, adverse perinatal outcome or any of the other variables studied. CONCLUSIONS: Tight glycemic control during pregnancy is recommendable to avoid progression of retinopathy. Attention should be given to the period after delivery where the tight regulation may be difficult to achieve. IDDM women should be encouraged to plan pregnancies early in life. PMID- 10830764 TI - Epidemiology of fetal death in Latin America. AB - BACKGROUND: To identify risk factors associated with fetal death, and to measure the rate and the risk of fetal death in a large cohort of Latin American women. METHODS: We analyzed 837,232 singleton births recorded in the Perinatal Information System Database of the Latin American Center for Perinatology and Human Development (CLAP) between 1985 and 1997. The risk factors analyzed included fetal factors and maternal sociodemographic, obstetric, and clinical characteristics. Adjusted relative risks were obtained, after adjustment for potential confounding factors, through multiple logistic regression models based on the method of generalized estimating equations. RESULTS: There were 14,713 fetal deaths (rate=17.6 per 1000 births). The fetal death risk increased exponentially as pregnancy advanced. Thirty-seven percent of all fetal deaths occurred at term, and 64% were antepartum. The main risk factors associated with fetal death were lack of antenatal care (adjusted relative risk [aRR]=4.26; 95% confidence interval, 3.84-4.71) and small for gestational age (aRR=3.26; 95% CI, 3.13-3.40). In addition, the risk of death during the intrapartum period was almost tenfold higher for fetuses in noncephalic presentations. Other risk factors associated with stillbirth were: third trimester bleeding, eclampsia, chronic hypertension, preeclampsia, syphilis, gestational diabetes mellitus, Rh isoimmunization, interpregnancy interval<6 months, parity > or =4, maternal age > or =35 years, illiteracy, premature rupture of membranes, body mass index > or =29.0, maternal anemia, previous abortion, and previous adverse perinatal outcomes. CONCLUSIONS: There are several preventable factors that should be dealt with in order to reduce the gap in fetal mortality between Latin America and developed countries. PMID- 10830765 TI - A follow-up study of birth outcome in users of pivampicillin during pregnancy. AB - BACKGROUND: Pivampicillin is a prodrug which is widely used in Scandinavian countries for oral antibiotic therapy. The pivaloyl moiety has a carnitine depleting effect, which has caused doubts about the safety of administering pivampicillin during pregnancy. The aim of the study was to evaluate the risk of congenital malformations in general, preterm delivery and low birth weight in users of pivampicillin. METHODS: Seven hundred and ninety-one women who had redeemed a prescription of pivampicillin during their first pregnancy from 1 January 1991 to 31 December 1996 were identified in the North Jutland Pharmaco Epidemiological Prescription Database. By linkage to the Danish Medical Birth Registry and Regional Hospital Discharge Registry we compared their birth outcomes (malformations, preterm delivery and low birth weight) with the outcomes in 7472 reference pregnancies on which the mother had not redeemed any prescription at all during pregnancy. RESULTS: The prevalence of malformations was 5.5% (11 cases) in offspring of 199 women who had used pivampicillin during the first trimester, and 5.6% (420 cases) in offspring of controls (OR: 0.95, 95% CI: 0.51-1.76). Furthermore, we did not find any significant risk of preterm delivery (OR: 0.75, 95% CI: 0.54-1.05) or low birth weight (OR: 0.93, 95% CI: 0.55-1.57). CONCLUSION: This study showed no increased risk of congenital malformations, preterm delivery or low birth weight in offspring of women who had redeemed a prescription for pivampicillin during pregnancy. PMID- 10830766 TI - Danish National IVF Registry 1994 and 1995. Treatment, pregnancy outcome and complications during pregnancy. AB - SUBJECT: Data from the compulsory Danish National IVF Registry from 1994 and 1995 regarding treatments, abortions and complications following assisted reproductive technologies. METHODS: Data were generated through registries and compared to pregnancies in Denmark in 1995. Those pregnancies that resulted in a delivery were compared to a matched control group. RESULTS: In 1994 and 1995 5219 women were treated in 9471 initiated cycles. The numbers increased over the period. The overall delivery rate per initiated cycle was 19%, egg donation 24%, IVF 20%, ICSI 16% and frozen egg replacement 10%. The rates increased over the period. The rate of spontaneous abortions was highest for ICSI (25%) and egg donation (27%). For IVF and ICSI the birth rates per transfer of 1 embryo was 13, 1%, 2 embryos 25, 4%, 3 embryos 25, 8% and 4 or more 3, 8%. Transfer of 2 embryos resulted in 75% singleton, 25% twin and 0.2% triplet deliveries. After transfer of 3 embryos the corresponding rates were 68%, 29% and 4%. No quadruplet deliveries occurred. Totally, 1.4% reported complications to the treatment, the most frequent being ovarian hyperstimulation syndrome. In the study group 5.8% of the women who gave birth were diagnosed with imminent abortion vs. 3.0% in the control group (OR 1.98, CI 1.41-2.78). CONCLUSIONS: Transfer of three embryos did not result in higher pregnancy rates as compared to transfer of two embryos. The first data from the Danish IVF Registry support data from other registries regarding treatment, pregnancy outcome and complications during pregnancy. PMID- 10830767 TI - Treatment with 2% clindamycin vaginal cream prior to first trimester surgical abortion to reduce signs of postoperative infection: a prospective, double blinded, placebo-controlled, multicenter study. AB - BACKGROUND: Bacterial vaginosis (BV) and intermediate flora is known risk-factor for postoperative infection after surgical termination of pregnancy. Vaginal application of 2% clindamycin cream is an efficacious treatment for BV, but it is not known whether preoperative administration of clindamycin cream might reduce the signs of post-abortion infection after surgical termination of pregnancy. AIM: To evaluate whether preoperative treatment with clindamycin cream might reduce the signs of post-abortion infection after legal abortion. DESIGN: Prospective, double-blinded, placebo-controlled, multicenter study. MATERIAL AND METHODS: Consecutive women attending for surgical termination prior to 11+4 gestational weeks were approached. We randomized participants to preoperative vaginal treatment with 2% clindamycin cream or placebo cream in a double-blinded fashion. At all visits vaginal smears were air dried on microscopy slides to be stored. The rate of postoperative pelvic infection according to our definition was the main outcome variable, the cure rates of BV and of intermediate flora were secondary outcome variables. RESULTS: Of 1655 enrolled women, 1102 were evaluable for analyses. Fifty-eight women developed signs of post-abortion infection. Preoperative treatment with clindamycin cream significantly (RR: 4.2, 95% C.I. 1.2-15.9) reduced the risk of post-abortion infection among women with abnormal vaginal flora (BV and intermediate flora). Treatment with clindamycin cream in women with normal lactobacilli flora did not demonstrate any difference compared to the non-treatment group. CONCLUSION: Preoperative treatment for at least three days with clindamycin cream significantly reduced the risk for developing signs of post-abortion infection only among women with preoperative abnormal vaginal flora (BV and intermediate flora). PMID- 10830768 TI - Apoptosis, cellular proliferation and expression of p53 in human uterine leiomyomas and myometrium during the menstrual cycle and after menopause. AB - BACKGROUND: Cell proliferation, apoptosis and expression of p53 proteins were studied in human uterine leiomyomas and myometrium during the menstrual cycle and after menopause. METHODS: Expression of ki-67 and p53 was analyzed by immunohistochemistry and by immunoblotting. Apoptosis was detected by in situ 3' end labelling of cells with DNA fragmentation. RESULTS: In both the proliferative and the secretory phases, ki-67 expression was higher in leiomyomas than in myometrium and both tissues showed higher expression in the secretory than in the proliferative phase. No difference in apoptotic index was observed between leiomyomas and myometrium or between the proliferative and secretory phases. After menopause, the expression of ki-67 as well as the apoptotic index was lower than in the proliferative and secretory phases and no significant difference between tissues was seen. Both leiomyomas and myometrium showed negative staining for p53. Immunohistochemical results regarding p53 were confirmed by Western blot. CONCLUSIONS: Our findings indicate that sex steroids influence the growth of leiomyomas by stimulating cell proliferation rather than by affecting apoptosis. The rate of cell proliferation is higher in fertile age than after menopause and appears to be enhanced under the influence of progesterone. PMID- 10830769 TI - Prevalence of menstrual cycle symptom cyclicity and premenstrual dysphoric disorder in a random sample of women using and not using oral contraceptives. AB - BACKGROUND: The prevalence of symptom cyclicity and premenstrual dysphoric disorder in randomly recruited samples of women has not been ascertained. We assessed this prospectively in 83 Icelandic women, aged 20-40 years, using and not using oral contraceptives (OCs). METHODS: A check list of 57 symptoms, divided into 7 symptom groups, was used to assess experiences. All symptoms and summarized symptom scores within symptom groups were tested for significant change between the follicular and the luteal phase of each menstrual cycle. When a symptom or a summarized symptom score was higher during the luteal phase compared to the follicular phase the cyclicity is labelled 'expected' cyclicity and the opposite 'unexpected' cyclicity. RESULTS: The women charted from 1-7 menstrual cycles each, with 66 of them charting 3-4 cycles. Cyclicity was never displayed by 9.6% of participants; 80.7% displayed expected cyclicity and 72.3% unexpected cyclicity at least once. Cyclicity in somatic symptoms was most often demonstrated by participants. Expected and unexpected cyclicity in summary symptom scores in well-being and in psychoemotional symptoms was very similar among participants. There was not much difference in cyclicity between non OC users and OC users. Fifty-one percent of the participants were self-defined with premenstrual syndrome with 2%-6% of them meeting the criteria of premenstrual dysphoric disorder. CONCLUSIONS: The normalcy of cyclicity found in this study might explain the high prevalence of premenstrual syndrome in many recall surveys. The importance of prospective daily ratings for establishing symptom cyclicity in women cannot be over emphasized. PMID- 10830770 TI - Laparoscopic colposuspension: a short term urodynamic follow-up and a three-year questionnaire-study. AB - BACKGROUND: The purpose of the study was to evaluate outcome, urodynamic correlates and adverse effects of laparoscopic colposuspension using polytetrafluoroethylene sutures. METHODS: Eighty-five consecutive women with primary stress urinary incontinence at one university hospital were included in this prospective non-controlled study. During videolaparoscopic surgery, two polytetrafluoroethylene sutures were placed on each side of the urethra and fixed to the Cooper ligaments. Pre- and postoperative clinical and urodynamic evaluations, including pad-test, were performed. A mailed questionnaire was used to evaluate cure-rate and complication-rate three years after surgery. RESULTS: At follow-up examination, we considered 62 of 76 women (82%) as being cured, ten (13%) improved, and four (5%) as being failures. The questionnaires were returned by 80 women; 41 (51%) considering themselves as cured and 31 (39%) improved, and eight women (10%) as unimproved or minimally improved. Clinical outcome was not associated with alterations in urethral functional length or in urethral closing pressure. Short preoperative urethral functional length was associated with failure (p=0.04).The incidence of new onset urge symptoms and of new onset recto/enterocele was 13% and 9% respectively. CONCLUSIONS: Laparoscopic colposuspension resulted in acceptable cure rate in short-, and medium long term evaluation. However, a decline in cure rate was observed. Cured women had significantly longer preoperative urethral functional length than women still leaking after surgery. PMID- 10830771 TI - Effectiveness of ultrasonographic parameters for documenting the severity of anatomic stress incontinence. AB - BACKGROUND: To assess the correlation between the severity of incontinence and some ultrasonographic parameters used to document the mobility of bladder neck. METHODS: The severity of the incontinence and mobility the bladder neck were evaluated by pad test and perineal ultrasonography respectively in 74 patients with pure anatomic stress incontinence. During perineal ultrasonography, distances from the bladder neck to the lower tip (A) and to the posterior midpoint of symphysis pubis (B) were measured at rest and during stress. The first bladder neck mobility was obtained by two caliper method (Mu). After superimposing the symphysis pubis of the frozen images, the bladder neck positions at rest and during stress were plotted on an x-y coordinate system by using computer programs and the corresponding X and Y values were calculated. The second bladder neck mobility was measured on this x-y coordinate system (Mc). Paired t-test and correlation analysis were used for statistical analysis of the data. RESULTS: The mean Mc was significantly higher than the mean Mu (p<0.01). Compared to the values at rest, the mean distance A did not change significantly; however, the mean distance B increased, and X and Y values decreased significantly during stress (p<0.01). The amount of urine leaked per hour had significant positive correlation with distance B-stress and Mc and significant negative correlation with Y-rest, X-stress and Y-stress (p<0.05-0.001). CONCLUSION: These data suggested that the bladder neck mobility was underestimated when it was measured by two caliper method and the absolute positions or mobility of the bladder neck measured by computer on an x-y coordinate system could effectively document the severity of the decreased support of bladder neck. PMID- 10830772 TI - Microlaparoscopy: a new approach to the reassessment of ovarian cancer patients. AB - BACKGROUND: The purpose of the work was to determine the feasibility and accuracy of microlaparoscopy as diagnostic method for the reassessment of ovarian cancer patients. METHODS: Eight patients scheduled for second-look laparoscopy who had undergone primary surgery for ovarian cancer followed by 6 cycles of chemotherapy were included in the study. Microlaparoscopy was performed using a 2.8 mm laparoscope followed by conventional 10-mm laparoscopy. Three additional 5-mm ancillary trocars were inserted to perform intraabdominal biopsies. Pelvic washings were performed in all cases. RESULTS: Microlaparoscopy was feasible in all cases and as accurate as conventional laparoscopy in seven cases. In one case the procedure was terminated before conventional laparoscopy because of positive biopsies at frozen section examination. There were no intra-operative complications related to microlaparoscopy. The median time from skin incision to the removal of the microlaparoscope was 47 minutes (range 30-70). CONCLUSION: Microlaparoscopy seems to be a safe, accurate, minimally invasive method and therefore we suggest its use as primary approach to the reassessment of ovarian cancer patients. PMID- 10830773 TI - Spontaneous rupture of unscarred uterus in early pregnancy--a rare entity. PMID- 10830774 TI - Neutrophils: molecules, functions and pathophysiological aspects. PMID- 10830775 TI - Dissociation of inflammatory and epithelial responses in a murine model of chronic asthma. AB - To study pathogenetic mechanisms in chronic asthma, we employed a novel experimental model that replicates characteristic features of the human disease. Chronic inflammation and epithelial changes, specifically localized to the airways, were induced by repeated exposure of systemically sensitized BALB/c mice to low mass concentrations of aerosolized ovalbumin for 6 weeks. The contribution of Th2 cytokine-driven inflammation to the development of airway lesions and hyperreactivity was assessed in cytokine-deficient mice. In interleukin-5 deficient animals, intraepithelial eosinophils and chronic inflammatory cells in the lamina propria of the airways were markedly decreased; however, these animals developed epithelial hypertrophy and subepithelial fibrosis comparable with that observed in sensitized wild type mice. Airway hyperreactivity to inhaled methacholine did not develop in interleukin-5-deficient mice. In contrast, interleukin-4-deficient mice exhibited no decrease in airway inflammation, but had significantly greater epithelial hypertrophy and subepithelial fibrosis, as well as exaggerated hyperreactivity to methacholine. We conclude that interleukin 5, but not interleukin-4, plays a central role in the development of chronic inflammation of the airways and the induction of airway hyperreactivity. Furthermore, chronic epithelial and fibrotic changes occur independently of interleukin-5 and are not required for the development of airway hyperreactivity. The dissociation between airway wall remodeling and airway hyperreactivity has important implications for therapeutic approaches to chronic asthma. PMID- 10830776 TI - Association of Mycobacterium leprae with human endothelial cells in vitro. AB - Endothelial cell infection by Mycobacterium leprae has long been described histologically in all types of leprosy and in some of the acute reactions occurring in this disease. Recent evidence from experimental lepromatous neuritis indicates that M. leprae colonizes endothelial cells of epineural blood vessels even in sites of minimal infection, suggesting that interaction between these cells and M. leprae may play an important role in the selective localization of this organism to peripheral nerve. To begin to study the mechanisms involved, we have examined the interaction between M. leprae and human umbilical vein endothelial cells (HUVEC) in vitro using light microscopy, scanning and transmission electron microscopy, and confocal laser scanning microscopy. When M. leprae were added to confluent monolayers of HUVEC, uptake increased slowly to a maximum at 24 hours. Maximal percentages of infected cells were similar at ratios of organisms:cell over a range of 25:1 to 100:1. The bacilli appeared to lie within membrane-bound vacuoles at all time points. The kinetics of association of M. leprae with HUVEC are much slower than has previously been observed with macrophages, possibly due to differences in the binding of M. leprae. Compared with other pathogens that infect endothelial cells, M. leprae also appear to be ingested more slowly, and to a more limited degree. The receptors involved in M. leprae binding to endothelial cells and the impact of intracellular infection by M. leprae on these cells remain to be determined. PMID- 10830777 TI - Ceramide induces cell death in the human prostatic carcinoma cell lines PC3 and DU145 but does not seem to be involved in Fas-mediated apoptosis. AB - Treatment of cells with synthetic C2-ceramide has been reported to induce apoptosis in several cell systems, and endogenously formed ceramide has been proposed to act as a second messenger, activating signaling pathways which contribute to the execution of apoptotic cell death after Fas ligation or tumor necrosis factor receptor-1 ligation. In this study, we examined the effect of exogenously administered C2-ceramide on the human prostatic carcinoma cell lines PC3 (Fas-sensitive) and DU145 (Fas-resistant). In both cell lines, C2-ceramide induced cell death in a dose-dependent manner, whereas a structural analog, C2 dihydroceramide, did not. The pan-caspase inhibitor zVAD-fmk did not prevent C2 ceramide-induced cell death but did prevent C2-ceramide-induced DNA fragmentation, indicating that apoptotic and non-apoptotic mechanisms are involved in C2-ceramide-induced death. Interestingly, cycloheximide prevented C2 ceramide-induced DNA fragmentation, indicating that ceramide-induced apoptosis in PC3 and DU145 requires new protein synthesis. In addition, because cycloheximide converts Fas-resistant DU145 to Fas-sensitive as assessed by DNA fragmentation, ceramide does not seem to play a major role in the Fas-mediated pathway in this cell line. We also determined the levels of endogenous sphingomyelin after Fas ligation in PC3. No decrease of sphingomyelin levels could be detected after Fas activation. We conclude that sphingomyelinase-generated ceramide does not play a role in Fas-mediated apoptosis in PC3, and that there are fundamental differences in the mechanisms of cell death induced by C2-ceramide and Fas ligation. PMID- 10830778 TI - Expression and tissue localization of membrane-types 1, 2, and 3 matrix metalloproteinases in rheumatoid synovium. AB - In vitro, membrane-type matrix metalloproteinases (MT-MMP) are known to activate the zymogen of MMP-2 (proMMP-2, progelatinase A), which is one of the key MMP in joint destruction in rheumatoid arthritis. In the present study, we examined the production and activation of proMMP-2, and the expression of MT1-MMP, MT2-MMP, and MT3-MMP, their correlation with proMMP-2 activation, and their localization in rheumatoid synovial tissue. Using sandwich enzyme immunoassay and gelatin zymography techniques, proMMP-2 production levels and activation ratios were found to be significantly higher in rheumatoid synovium compared with normal synovium (p < 0.01). Quantitative RT-PCR analyses demonstrated that MT1-MMP and MT3-MMP were expressed in all rheumatoid synovial tissue (30 of 30 cases), but that the mean expression level of MT1-MMP was approximately 11-fold higher than MT3-MMP. Significant correlation was found between the mRNA expression level of MT1-MMP and the activation ratio of proMMP-2 (p < 0.01). In situ hybridization indicated that the hyperplastic lining cells of rheumatoid synovium expressed MT1 MMP. Immunohistochemistry demonstrated that MT1-MMP was co-localized with MMP-2 and with a tissue inhibitor of metalloproteinase-2, and was mainly located in the rheumatoid synovial lining cells. In situ zymography of rheumatoid synovium showed gelatinolytic activity, predominantly in the lining cell layer. This activity was blocked when incubated with BB94, a specific MMP inhibitor. These results demonstrate that MT1-MMP plays an important role in the activation of proMMP-2 in the rheumatoid synovial lining cell layer, and suggest that its activity may be involved in the cartilage destruction of rheumatoid arthritis. PMID- 10830779 TI - Correlation of p16 hypermethylation with p16 protein loss in sporadic gastric carcinomas. AB - Hypermethylation of p16 has been detected frequently in a variety of cancer cells and is known to repress the level of p16 transcription. In human gastric carcinoma (GC) cells, p16 protein loss has often been detected, but genetic alterations of p16 are infrequent. To investigate the molecular mechanism of p16 gene inactivation in gastric carcinogenesis, we examined the methylation status of p16 in GC using methylation-specific PCR. Thirty-seven of eighty-eight (42%) GC showed p16 hypermethylation. Immunohistochemical analysis of 41 cases of GC showed a complete loss of p16 immunoreactivity in 19 of 22 (86%) methylation positive cases, but in only 2 of 19 (11%) methylation-negative cases. Of 88 GC, 21 cases were previously identified as having microsatellite instability (MSI). Interestingly, 13 of 21 (62%) MSI-positive tumors and 24 of 67 (36%) MSI-negative tumors had hypermethylation on p16. The relatively high frequency of hypermethylation on p16 and the strong correlation between the immunoreactivity and methylation patterns suggest that methylation is an important mechanism for p16 gene inactivation in GC. PMID- 10830780 TI - PDGF-mediated chemoattraction of hepatic stellate cells by bile duct segments in cholestatic liver injury. AB - The accumulation of myofibroblasts and fibrosis around proliferating bile ducts in cholestatic liver disease has been attributed to the proliferation and phenotypic modulation of portal fibroblasts, whereas the contribution of hepatic stellate cells remains uncertain. There is increasing evidence to indicate that bile ducts may stimulate chemoattraction of hepatic stellate cells (HSC). In the present study, we undertook dynamic tests to examine such a possibility and to investigate the role of two potential mediators: platelet-derived growth factor BB (PDGF-BB) and endothelin-1. Cholestasis was induced by bile duct ligation in rats. HSC were isolated from normal rats and culture activated into myofibroblasts expressing PDGF-beta receptors. Migration of myofibroblastic HSC was investigated in a Transwell chemotaxis filter assay. As compared with basal conditions, PDGF-BB (100 microg/l) and endothelin-1 (10(-8) M) induced a 3-fold and 1.7-fold increase in HSC migration, respectively. Bile duct segments isolated from cholestatic rats triggered a 3-fold increase in migration. This stimulation was significantly more potent than that observed in the presence of normal bile ducts. It was inhibited by neutralizing anti-PDGF antibodies and by STI571 PDGF receptor tyrosine kinase inhibitor, by 60% and 85%, respectively, whereas Bosentan, an endothelin receptor antagonist, had no significant inhibiting effect. In bile duct segments from cholestatic rats PDGF-B chain mRNA was detected at higher levels than in controls, whereas PDGF-BB was immunolocalized in bile duct epithelial cells. The results indicate that chemotaxis of HSC towards bile duct structures may contribute to the development of periductular fibrosis in cholestatic disorders, and that PDGF-BB is the major mediator in this process. In addition, anti-liver fibrogenic properties of STI571 are suggested by potent inhibition of myofibroblastic HSC function. PMID- 10830781 TI - Clonality and genetic divergence in multifocal low-grade superficial urothelial carcinoma as determined by chromosome 9 and p53 deletion analysis. AB - Multifocality and recurrence are clinically important features of urothelial carcinomas of the urinary bladder. Recent molecular genetic studies have suggested that multifocal urothelial carcinomas are monoclonally derived from an identical transformed progenitor cell. However, most of these studies investigated advanced and poorly differentiated tumors. The study presented focuses on early papillary tumors, including 52 superficial well-differentiated multifocal and recurrent bladder carcinomas from 10 patients. Microdissection separating urothelium from stromal cells was considered essential to obtain pure tumor cell populations. Genetic analysis was carried out by applying two different methods. Dual color fluorescence in situ hybridization (FISH) with centromeric probes for chromosomes 9 and 17 and gene-specific probes for chromosome loci 9q22, 9p21, and 17p13 was carried out in parallel to loss of heterozygosity (LOH) analyses applying 5 microsatellite markers on these chromosomes. Overall, deletions on chromosome 9p were found in 47 tumors (90%), at chromosome 9q in 36 tumors (69%) and at chromosome 17p in 3 tumors (6%). There was a very high correlation of the results between FISH and LOH analysis. Ten early superficial papillary tumors showed deletion of chromosome 9p without deletion of 9q, suggesting 9p deletions as a very early event in the development of papillary urothelial carcinoma. Although in four patients, all investigated tumors showed identical genetic alterations and one patient showed no genetic alterations at the loci investigated, in five patients, two or more clones with different deletions were found. In four of these patients, the results are compatible with clonal divergence and selection of different cell subpopulations derived from a common progenitor cell. However, in one patient different alleles in two markers at chromosome 9 were deleted, favoring an independent evolution of two recurring tumor cell clones. In summary, we could show that there is considerable genetic heterogeneity in early multifocal and recurring urothelial carcinoma and demonstrated the occurrence of two independent clones in at least one patient as an indicator of possible initial oligoclonality of bladder cancer. PMID- 10830782 TI - Plasma Escherichia coli beta-galactosidase as a marker of tumor burden and response to experimental anti-neoplastic therapy in nude mice xenografted with lacZ transduced human tumor cells. AB - Genetic labeling of tumor cells with the Escherichia coli lacZ reporter gene, encoding the enzyme beta-galactosidase, is widely used for histochemical detection of micrometastases in mice. Recently, we have developed a novel, highly sensitive and specific immunocapture chemiluminescence assay for the quantitation of E. coli beta-galactosidase. This assay achieved a detection limit of 0.01 mU of E. coli beta-galactosidase per milliliter, and 97% signal recovery of purified enzyme added to mouse plasma. LacZ transduced MDA-MB-231 BAG human breast cancer cells grown in vitro released soluble beta-galactosidase into the culture medium, and the concentration found correlated with cell density. Growth of the same cells in nude mice produced readily measurable levels of E. coli beta galactosidase enzyme activity in host plasma and a highly significant correlation could be demonstrated between the size of primary tumor xenografts and the host plasma level of E. coli beta-galactosidase activity. When mice bearing MDA-MB-231 BAG tumor xenografts were treated intravenously with a single injection of doxorubicin (5 mg/kg), the mean tumor volume after 16 days was reduced 4-fold in the group of doxorubicin-treated mice compared with saline-treated control mice, and the mean level of plasma E. coli beta-galactosidase was correspondingly reduced 3.8-fold in the doxorubicin-treated mice compared with control mice. Sensitive and specific measurement of soluble E. coli beta-galactosidase in blood, using an immunocapture chemiluminescence assay, thus provides objective assessment of tumor burden in mice xenografted with lacZ transduced human tumors. This assay may have important applications as a tool for determining the efficacy of new experimental anti-tumor agents. PMID- 10830783 TI - Induction of chromosomal aberrations and growth-transformation of lymphoblastoid cell lines by inhibition of reactive oxygen species-induced apoptosis with interleukin-6. AB - Etiological evidence, indicating the relationships between the onset of malignant lymphoma and pre-existing chronic inflammation, has been accumulated. For the autonomous growth of malignant tumor, genetic lesions, such as chromosomal aberrations, amplification of oncogenes, and mutations of genes involved in the cell cycle regulation, must be essential. However, how the inflammation promotes the accumulation of genetic lesions and induces the autonomous growth of lymphoid cells remains unclear. Reactive oxygen species released by polymorphonuclear leukocytes and macrophages are factors causing DNA damage in the foci of inflammation, and thus could play a role in lymphomagenesis. The xanthine/xanthine oxidase (X/XOD) system produces a mixture of hydrogen peroxide and superoxide anion extracellularly, and thus serves as an in vitro source of reactive oxygen species. Cell death of lymphoblastoid cell lines (LCLs) was induced with X/XOD treatment in a dose-dependent manner. DNA fragmentation, which is the characteristic feature of apoptosis, was observed in LCLs at 4-8 hours after X/XOD treatment. Among cytokines such as interleukin-6 (IL-6), IL-10, and interferon-gamma, only pretreatment with IL-6 gave LCLs the resistance to X/XOD induced cell death in a dose-dependent manner. The proportion of apoptotic cells in X/XOD-treated LCL culture was decreased with IL-6 pretreatment by quantification with flow cytometric analysis. Treatment of LCLs with IL-6 for 48 hours up-regulated bcl-2 mRNA expression. Furthermore, the LCLs repeatedly treated with X/XOD and cultured with or without IL-6 showed many more structural abnormalities of chromosomes than those without X/XOD treatment. Colony forming efficiency of X/XOD-treated LCLs with IL-6 was significantly higher than those without IL-6, and even relatively higher than LCLs without X/XOD treatment. IL-6 could support the survival of non-neoplastic B cells and accelerate the malignant transformation of B lineage cells in inflammatory lesions. PMID- 10830784 TI - Replenishment of glutathione levels improves mucosal function in experimental acute colitis. AB - Because reactive oxygen species (ROS) have been implicated as mediators of inflammatory bowel disease (IBD), the purpose of the present work was to determine the functional role of mucosal GSH in the trinitrobenzenesulfonic acid in 50% ethanol (TNBS+ethanol)-induced colitis in rats. Mucosal samples were taken to evaluate the temporal relationship between the extent of injury, the levels of glutathione (GSH) during acute colitis induced by TNBS+ethanol, and the effect of N-acetylcysteine (NAC) administration. In vitro assays revealed the interaction of TNBS with GSH leading to the almost instantaneous disappearance of GSH, while the reductive metabolism of TNBS by GSSG reductase generated ROS. Mucosal samples from TNBS+ethanol-treated rats indicated a direct correlation between GSH depletion and injury detected as soon as 30 minutes after TNBS+ethanol administration that persisted 24 hours post treatment. Although, short term depletion of mucosal GSH per se by diethylmaleate did not result in mucosal injury, the oral administration of NAC (40 mM) 4 hours after TNBS+ethanol treatment increased GSH stores (2-fold), decreasing the extent of mucosal injury (60-70%) examined at 24 hours post treatment. However, an equimolar dose of dithiothreitol failed to increase GSH levels and protect mucosa from TNBS+ethanol induced injury. Interestingly, GSH levels in TNBS+ethanol-treated rats recovered by 1-2 weeks, an effect that was accounted for by an increase of gamma glutamylcysteine synthetase (gamma-GCS) activity due to an induction of gamma-GCS heavy subunit chain mRNA. Thus, TNBS promotes two independent mechanisms of injury, GSH depletion and ROS generation, both being required for the manifestation of mucosal injury as GSH limitation renders intestine susceptible to the TNBS-induced ROS overgeneration. Accordingly, in vivo administration of NAC attenuates the acute colitis through increased mucosal GSH levels, suggesting that GSH precursors may be of relevance in the acute relapse of IBD. PMID- 10830785 TI - Metallothionein inhibits myocardial apoptosis in copper-deficient mice: role of atrial natriuretic peptide. AB - Dietary copper restriction causes heart hypertrophy in animal models. Several studies have indicated that this cardiomyopathy is mediated by oxidative stress. Metallothionein (MT), a low molecular weight and cysteine-rich protein, functions in protecting the heart from oxidative injury. We therefore used a cardiac specific MT-overexpressing transgenic mouse model to test the hypothesis that MT inhibits copper deficiency-induced heart hypertrophy. Dams of both transgenic pups and non-transgenic littermates were fed a copper-adequate or copper deficient diet, starting on the fourth day post-delivery, and the weanling mice were continued on the dams' diets until they were killed. Heart hypertrophy developed in copper-deficient pups by the fourth week of the combined pre- and post-weaning feeding and aggressively progressed until the end of the experiment (6 weeks). MT overexpression did not prevent the occurrence of heart hypertrophy, but inhibited the progression of this cardiomyopathy, which correlated with its suppression of cardiac lipid peroxidation. Corresponding to the progression of heart hypertrophy, myocardial apoptosis and atrial natriuretic peptide (ANP) production in the left ventricle were detected in non-transgenic copper-deficient mice; these effects were significantly suppressed in transgenic copper-deficient mice. Measurement of apoptosis by TUNEL assay and Annexin V-FITC confocal microscopy in primary cultures of cardiomyocytes revealed that ANP was largely responsible for the myocyte apoptosis and that MT inhibited ANP-induced apoptosis. The data clearly demonstrate that elevation of MT in the heart inhibits oxidative injury and suppresses the progression of heart hypertrophy in copper deficiency, although it does not block its initiation. The results suggest that MT inhibits the transition from heart hypertrophy to failure by suppressing apoptosis through inhibition of both cardiac ANP production and its apoptotic effect. PMID- 10830786 TI - Protective role of interleukin-1 in mycobacterial infection in IL-1 alpha/beta double-knockout mice. AB - To understand the role of the proinflammatory cytokine interleukin-1 (IL-1) in mycobacterial inflammation, IL-1 alpha/beta double-knockout (KO) mice were produced. These mice were infected with either Mycobacterium tuberculosis H37Rv by the airborne route using an airborne infection apparatus, and their capacities to control mycobacterial growth, granuloma formation, cytokine, and nitric oxide (NO) production were examined. The IL-1 alpha/beta mice developed significantly larger (p < 0.01) granulomatous, but not necrotic, lesions in their lungs than wild-type (WT) mice after infection with H37Rv. Inflammatory lesions, but not granulomas, were observed in spleen and liver tissues from both IL-1 alpha/beta KO and wild-type mice. Granulomatous lesion development in IL-1 alpha/beta KO mice was not significantly inhibited by treatment with exogenous recombinant IL-1 alpha/beta. Compared with wild-type mice, splenic IFN-gamma and IL-12 levels were within the normal range. NO production by cultured alveolar macrophages from IL-1 alpha/beta KO mice was lower than in wild-type mice but were increased by the addition of recombinant IL-1 alpha/beta. Our data clearly indicate that IL-1 is important for the generation of early-phase protective immunity against mycobacterial infection. PMID- 10830787 TI - Islet abnormalities associated with an early influx of dendritic cells and macrophages in NOD and NODscid mice. AB - In the nonobese diabetic (NOD) mouse model for type 1 diabetes, the inflammatory infiltration of islets starts with an influx of dendritic cells (DC) and macrophages (Mphi) at approximately 4 weeks of age. Around this time, NOD mice show endocrine abnormalities, indicated by a transient hyperinsulinemia that lasts until 8 weeks of age. Subsequently, they develop abnormally large islets of Langerhans, here designated as "mega-islets." NODscid mice, which lack functional lymphocytes, also exhibit transient hyperinsulinemia, but to a lesser extent. First, to determine the role of lymphocytes in the morphological islet abnormalities, we compared 6-week-old (prediabetic) NOD and NODscid females regarding mega-islet development and accumulation of antigen-presenting cells (APC), particularly CD11c+ DC and ERMP23+ Mphi. In NODscid mice, early APC infiltration and mega-islets were present, but less marked compared with NOD mice, thus suggesting a role of lymphocytes in mega-islet formation. In both NOD and NODscid mice, the APC infiltration was predominantly found around the mega islets, suggesting a relationship between both parameters. Second, to analyze the role of beta-cell hyperactivity in mega-islet formation, we studied the effect of short-term prophylactic insulin treatment on these parameters. Prophylactic insulin treatment decreased the percentages of mega-islets in both NOD and NODscid mice, indicating that beta-cell hyperactivity is also involved in mega islet formation. In conclusion, mega-islet formation in mice with the NOD genetic background takes place under the influence of both beta-cell hyperactivity and leukocytes. PMID- 10830788 TI - Initial intermittent microwave irradiation for fluorescence in situ hybridization analysis in paraffin-embedded tissue sections of gastrointestinal neoplasia. PMID- 10830789 TI - New clinical trials of angiotensin blockade in cardiovascular disease: foreword. PMID- 10830790 TI - Worldwide trends and shortcomings in the treatment of hypertension. AB - Despite advances in the availability and quality of antihypertensive medications and the institution of national programs to educate people about the seriousness of hypertension, much remains to be done. The percentages of people who are aware of their disease, are being treated for it, and are achieving adequate control of their blood pressure (BP) are disconcertingly low worldwide; surveys in numerous countries have consistently shown that fewer than 60% of affected people are being treated for hypertension and fewer than 30% have their BP adequately controlled. Reluctance on the part of many physicians to use antihypertensive therapy aggressively may be an impediment to improving these numbers. Further clinical trials are needed to assess the extent to which BP could be lowered safely and effectively with both single-agent and combination therapies. PMID- 10830791 TI - The role of the AT2 receptor in hypertension. AB - The renin-angiotensin system (RAS) is integrally involved in maintaining the healthy body's hemodynamic status. It is also involved in many pathogenic situations. Angiotensin II (Ang II) is the major effector hormone of this system. Ang II subtype 1 receptor blockers (ARB), like angiotensin-converting enzyme (ACE) inhibitors, modulate the potent vasoconstricting and growth-promoting effects of Ang II. Thus, it is reasonable to assume that ACE inhibitors and ARB provide similar benefits in patients with hypertension and other diseases. There are salient differences, however, in that ARB antagonize Ang II at its AT1 receptor subtype but spare its AT2 receptor subtype, which has unique-and largely oppositional- effects on the blood vessels, kidneys, and adrenals. ACE inhibitors decrease the amount of Ang II available to its AT1 and AT2 receptors alike without totally suppressing its formation. This article reviews recent findings about the role of the AT2 receptor in both health and disease and the actions of ARB mediated by this receptor. PMID- 10830792 TI - Treating high-risk hypertensive patients. AB - Successful treatment of hypertension entails not only normalizing high blood pressure, but also addressing the associated risk factors that increase the likelihood of cardiovascular morbidity and mortality. Hypertension often occurs in a setting of insulin resistance, hyperinsulinemia, dyslipidemia, and a prothrombotic state. A number of epidemiologic studies have shown that the clustering of these abnormalities is associated with increased risk of cardiovascular morbidity and mortality. Therefore, it is rational to direct therapy at moderating these risk factors as well as at lowering blood pressure in hypertensive patients. This is particularly important in patients with comorbidities such as diabetes, cardiovascular disease, or renal insufficiency. Many physicians prescribe only diuretics and beta-blockers, agents that have demonstrated efficacy in long-term randomized controlled trials. However, this approach does not consider the potential benefits of newer agents for which long term outcome data are not yet available. The ongoing Valsartan Antihypertensive Long-term Use Evaluation (VALUE) trial, in which the angiotensin II subtype 1 receptor blocker valsartan is compared with the third-generation calcium channel blocker amlodipine, should provide important evidence on the long-term efficacy of these newer agents. A unique feature of VALUE is that it is specifically enrolling into the only current trial, now under way, hypertensive men and women at a relatively high risk for a cardiovascular event to determine the benefits of complete blockade of angiotensin II beyond those of the control of blood pressure. PMID- 10830793 TI - Heart failure: future treatment approaches. AB - Large-scale clinical trials of vasodilators with nitrates and hydralazine and with angiotensin-converting enzyme (ACE) inhibitors in the 1980s and early 1990s provided the first credible evidence that medical therapy can prolong survival in patients with chronic heart failure (CHF). Moreover, patients treated with ACE inhibitors required fewer hospitalizations for worsening heart failure (HF). Nonetheless, the prognosis in patients with HF remains bleak, and better therapies are urgently needed. Recently, beta-blockers and spironolactone have been shown to reduce mortality when added to ACE inhibitors, diuretics, and digoxin. Digoxin has a neutral effect on overall mortality but does reduce the rate of hospitalization. Angiotensin II receptor blockers (ARB) inhibit the AT1 angiotensin receptor, which mediates the deleterious effects of the renin angiotensin system, and may provide advantages over ACE inhibitors or advantages when used in combination with ACE inhibitors. Newer drugs that interfere with other mechanisms that contribute to progression of heart failure are also under study. As new therapies prove effective in large populations, they lead to a mandate for polypharmacy. The long-term solution to this clinical problem is to develop sensitive and reliable markers that can predict response in individual patients or monitor effectiveness of therapy. PMID- 10830794 TI - Effect of cetrorelix on sperm morphology during migration through the epididymis in the cynomolgus macaque (Macaca fascicularis). AB - The importance of the cynomolgus monkey as a model for human reproductive medicine prompted this examination of epididymal sperm morphology. Computer-aided sperm morphological analysis was used for the first time to provide morphometric data on sperm heads as they traversed the epididymal duct of Macaca fascicularis. The duct was divided into six regions, starting close to the testis (proximal) and ending close to the vas deferens (distal). To determine the androgen dependence of the changes, one group of animals received a GnRH-antagonist (Cetrorelix, Asta Medica, Frankfurt, Germany) to induce testicular regression and lower epididymal androgens, while a control group received only vehicle. Epididymides were removed 16 and 25 days after treatment, and sperm heads were analysed by a computer-assisted morphometric analyser. Cluster analysis revealed swollen sperm head cells in proximal regions 1 and 2 of the epididymis, but fewer such forms distally. Normal head shapes became the majority in region 4 and these underwent a gradual but statistically significant decrease in size (area, perimeter, length, width) and shape as they reached the distal regions. In the animals given Cetrorelix, sperm with swollen heads were found more distally than in the controls, although they were also never present in the distal cauda (region 6). Normal heads still became predominant in region 4 after 16 days treatment, and in region 6 after 25 days. The normal forms in the cauda epididymidis of treated animals were significantly larger than cells from control animals. We conclude that epididymal sperm maturation in the monkey is characterised by both a loss of sensitivity to distortion on air-drying, and by a decrease in sperm head size. The former, but not the latter, is attained by sperm in androgen-deficient epididymides from GnRH-antagonist-treated monkeys. PMID- 10830795 TI - Hormonal correlates of reproductive seasonality in wild female hanuman langurs (Presbytis entellus). AB - To date, it is not known whether the seasonal occurrence of sexual behavior and mating in free-ranging Hanuman langurs at Ramnagar, Nepal, is correlated with seasonal changes in female ovarian function, and, if so, which factor(s) triggers the onset of the reproductive period. Using noninvasive fecal hormone analysis in combination with behavioral observations, this study was carried out to: 1) investigate and characterize seasonal patterns of ovarian cyclicity and timing of conception in wild langur females living in a highly seasonal habitat; and 2) examine the relationship between seasonal patterns of ovarian cyclicity, behavioral estrus, and female physical condition. Behavioral data and fecal samples were collected during a total period of 14 months from nine females living in a multi-male group. Physical condition of the females was assessed monthly by visual inspection, using a seven-fold scale. Ovulatory cycles and timing of conceptions were identified by the measurement of immunoreactive pregnanediol glucuronide (iPdG) in extracted feces. Hormone profiles in individual females revealed a clearly seasonal distribution in the occurrence of ovulatory cycles, which were restricted to the period from July to October. The distribution of female estrus behavior showed a similar seasonal pattern, and in total 88.2% of all estrus periods observed in the focal females were accompanied by ovulation. Onset of ovarian cycles as well as mating activities were strongly correlated with the onset of the rainy season. Females conceived, on average, in their second ovulatory cycle (pregnancy length: 211.6 +/- 3.4 days), with timing of conception being confined to the months when animals showed an improved physical condition. Collectively the present data clearly suggest that in seasonally-breeding langurs at Ramnagar, ecological conditions (rainfall, food availability, and quality) influence the onset of ovulations and timing of conceptions. PMID- 10830796 TI - Phylogenetic analysis of twinning in callitrichinae. AB - The callitrichines are known for twinning and for a communal rearing system in which all or most group members help care for the offspring. The origin of twinning has been the subject of much speculation. In this study predictions from earlier hypotheses are tested on the basis of two alternative phylogenetic trees. From this analysis we infer that helping behavior and male care preceded the origin of twinning, and that these traits did not coevolve with, but might have been important prerequisites for twinning in callitrichines. Small body size does not necessarily result in twinning, although it might still have been a prerequisite for its evolution. Gum feeding was an ecological change which evolved along with twinning. If nutrition was a limiting factor in the number of off-spring produced, then the use of a new feeding resource could have been crucial for the origin of twinning in callitrichines. According to one of the two alternative solutions inferred by the total evidence tree, and in accordance with the morphological tree, semi-annual breeding appears in the marmosets together with specialization in gum feeding. The fact that gums are available for these monkeys all year may have facilitated semiannual breeding. We suggest that the exploitation of gums as a feeding resource could have been the decisive factor in the increase of the reproductive rate by twinning and by semi-annual breeding. PMID- 10830797 TI - Feeding tactics in juvenile rhesus monkeys (Macaca mulatta). AB - In a group of rhesus monkeys, feeding tactics of juveniles were studied in a competitive situation in which food presentation had been modified, and where food was supplied in a feeding trough six times a day. Juvenile offspring of high ranking mothers remained longer in the feeding area gathering food. In contrast, juvenile offspring of low-ranking mothers went less often to the feeding area, primarily to collect food. Low-ranking juveniles also gathered the food faster than did dominants and used longer feeding bouts. High-ranking juveniles interrupted their feeding more spontaneously than did subordinate ones. Low ranking juveniles did so more often as a result of avoiding and being startled. Juvenile males entered the feeding area to satisfy their food requirements more often than females, but were also aggressively expelled more than females. No relation was found between age in months of juveniles and any feeding parameters or causes of interruption. Nor were significant differences found between young and old juveniles. Juvenile rhesus appear to use different tactics in accessing food according to their sex and to their mothers' dominance rank. PMID- 10830798 TI - Vertebrate predation by sympatric tamarins, Saguinus mystax and Saguinus fuscicollis. AB - Vertebrate predation was examined in sympatric moustached tamarins (Saguinus mystax) and saddle-back tamarins (Saguinus fuscicollis) in the Amazon rainforest of northeastern Peru. Both species prey on frogs and lizards, and very rarely on nestling birds. As a result of divergent foraging strategies, S. mystax primarily exploited frogs at higher strata of the forest, while S. fuscicollis predominantly preyed on reptiles in the lower strata and on the ground. This difference may strengthen the niche differentiation between these two tamarin species that exists with regard to other prey. PMID- 10830799 TI - The state of the art in cleft palate surgery. AB - This update focuses on current practices and controversies in surgical repair of the hard and soft palate posterior to the alveolus. Our current understanding of the advantages and disadvantages of presurgical active and passive manipulation of the hard palate shelves including the use of periosteoplasty is reviewed. The evolution of the multiple methods of repair of the hard and soft palate is given in a historical context along with a discussion of the concerns about timing of palate repair. PMID- 10830800 TI - Management of alveolar cleft bone grafting--state of the art. AB - Bone grafting of the alveolus is an essential step in the reconstruction of the orofacial cleft deformity. Secondary grafting with iliac marrow consistently produces trabecular bone to unify the maxilla and provide odontogenic support. It requires preoperative maxillary alignment, well designed mucoperiosteal flaps, and good oral hygiene to be optimally successful. Its high success rate currently makes it the preferred approach at most centers. Primary grafting with rib results in a unified maxilla, eliminates the oronasal fistula, and does not adversely affect midfacial growth. It assists in preventing maxillary segmental collapse, particularly in the bilateral cleft patient. Whether it can produce enough alveolar bone to support long-term odontogenic needs awaits further clinical documentation in the limited numbers of centers that routinely perform this procedure. PMID- 10830801 TI - Repairing the cleft lip nasal deformity. AB - OBJECTIVE: The repair of the cleft lip nose and nasal deformity remains a challenging endeavor for reconstructive surgeons. Psychosocially, this complex, multifaceted deformity significantly stigmatizes the patient. Numerous techniques have been advocated by multiple authors for the treatment and reconstruction of these deformities, usually requiring serial staged reconstructions. METHOD: Described is our technique for early primary repair of the cleft lip nasal deformity. The use of multiple suspension sutures to repair the nasal defect facilitates the repair of even very wide cleft lips. CONCLUSIONS: These maneuvers provide an aesthetic and functional repair of the nasal defect in conjunction with the lip repair. Long-term results have minimized the need for surgical revision. PMID- 10830802 TI - A method of "unilateral operation" for early repair of unilateral complete cleft palate. Preliminary report. AB - OBJECTIVE: This article describes a method of "unilateral operation" and the preliminary results of a group of patients with unilateral complete cleft palate undergoing the operation at early age. DESIGN: The "unilateral operation" consists of four relaxation maneuvers. After all of the four maneuvers have been performed on the deformed side of an unilateral complete cleft palate, the deformed side can be moved posteriorly and medially to contact with the normal side. Then the cleft can be closed without tension. RESULTS: From 1995 to 1998, 19 cases of unilateral complete cleft palate were repaired with this method at 5 12 months of age. Postoperatively, there were no deaths nor dehiscences. Under the care and guidance of an experienced speech pathologist, 15 of 17 of these children have normal vocal quality at 1-2 years of age. CONCLUSIONS: The "unilateral operation" is a rational, adequate, and safe method for early repair of unilateral complete cleft palate. It's design addresses four principles. First, operating only on the deformed side of a unilateral complete cleft palate leaves the normal side unperturbed. Second, complete relaxation of the deformed side is achieved before closing the cleft. Third, in comparison with conventional procedures, which operate on both sides of the palate, this method has the advantage of less surgical trauma, less blood loss, and shorter time of operation. Fourth, all of these advantages are beneficial to early cleft palate repair, which is an important factor in achieving good speech. PMID- 10830803 TI - Nasalance distance and ratio: two new measures. AB - OBJECTIVES: Mean nasalance in speakers with perceptually normal nasal resonance can differ in magnitude considerably. In addition, categorizations of speech based on nasalance scores may not agree with perceptual judgments. To overcome this limitation, we evaluated two new simple measures derived from mean nasalance data: the nasalance distance (range between maximum and minimum nasalance) and the nasalance ratio (minimum nasalance divided by maximum nasalance). SETTING: Department of Oral and Maxillofacial Surgery, University of Technology, Munich, Germany. SUBJECTS: The sample consisted of 133 cleft lip and palate patients with normal nasal resonance or varying degrees of hypernasality. PROCEDURES: Oral and nasal acoustic measurements were made using the NasalView system. Nasalance distance and nasalance ratio were calculated for five non-nasal and three nasal sentences from the modified Heidelberg Rhinophonia Assessment Form. RESULTS: Optimum cutoffs were derived from receiver-operating characteristics. Results for the sentence stimuli ranged from 64.4% to 89.6% sensitivity and from 91.2% to 94.1% specificity. When the analysis was limited to only one nonnasal and one nasal sentence, results ranged from 79.7% to 87.5% sensitivity and from 88.2% to 97.1% specificity. CONCLUSIONS: We conclude that the two new measurements are valuable in routine clinical examinations. Nasalance distance and ratio derived from sentence stimuli are two useful and easily applicable measures that can be used to supplement the nasalance mean value. PMID- 10830804 TI - Are infants with orofacial clefts at risk for insecure mother-child attachments? AB - OBJECTIVE: Several risk factors in the early lives of children with clefts are believed to interfere with their development of secure attachments to parents; however, this possibility has rarely been studied empirically. This study compared 12- and 24-month attachment classifications of infants with cleft palate (CP), infants with cleft lip and palate (CLP), and a comparison group of unimpaired infants (COMP). METHOD: Twenty-two CP infants, 24 CLP infants, and 61 matched COMP infants were assessed at 12 and 24 months of age in an urban children's hospital. At both visits, mothers and infants participated in the Strange Situation, which was videotaped and subsequently coded for patterns of attachment behavior. RESULTS: CP infants displayed a lower rate of 12-month attachment security than infants in the CLP or COMP groups. By 24 months, no diagnostic group differences in attachment classification were found. Stable 12- to 24-month attachment classifications were less likely in the CP group (36.3%) than in the COMP (62.3%) group. CP infants who were insecure at 12 months were more likely to become secure by 24 months than were CLP or COMP group infants. CONCLUSIONS: In contrast to previous theory and clinical speculation, the facial appearance of infants with CLP does not appear to affect the early mother-infant relationship adversely. The infancy period is marked by attachment instability for infants with CP, who demonstrated lower-than-expected rates of security at 12 months. However, these problems resolved in nearly all cases by 24 months of age. Most infants with clefts emerged from the first 2 years of life with secure maternal attachments. PMID- 10830805 TI - A comparison of the effects of Veau-Wardill-Kilner palatoplasty and furlow double opposing Z-plasty operations on eustachian tube function. AB - OBJECTIVE: The purpose of this study was to compare the effects of Veau-Wardill Kilner (VWK) two-flap palatoplasty and Furlow double-opposing Z-plasty operations on eustachian tube (ET) opening in patients with cleft palate (CP). DESIGN: Twenty-six patients underwent CP repair between 1993 and 1997. VWK and Furlow operations were used in 13 patients each. All patients were examined by the otorhinolaryngologist before and after the operations; mean follow-up time was 27.3 months. Secretory otitis media (SOM), as identified by tympanograms, was the comparison criterion used. RESULTS: The prevalence of SOM was 69% (n = 18) in all CP patients preoperatively. Among those with preoperative SOM, we did not find any statistically significant difference in the frequency of postoperative SOM between the two techniques. CONCLUSION: This study suggests that there is no difference in postoperative ET function between patients undergoing either VWK or Furlow operations, in spite of the differences in muscle positioning in these operations. We discuss the implications of this finding in relation to surgical anatomy and histology of the soft palate muscles. PMID- 10830806 TI - Prevalence of intranasal ectopic teeth in children with complete unilateral and bilateral cleft lip and palate. AB - OBJECTIVE: The objective of this study was to determine the prevalence of ectopic eruption of intranasal teeth. DESIGN: This was a retrospective study, where records of children with repaired cleft lip and palate were analyzed. SETTING: The study was conducted at a large craniofacial center, the Hospital for Rehabilitation of Craniofacial Anomalies, USP, Bauru, Sao Paulo, Brazil. SUBJECTS: The sample consisted of 815 records from patients with bilateral cleft lip and palate (BCLP) and 1,495 records from patients with unilateral cleft lip and palate (UCLP). The age of the subjects was 5 to 10 years old and the groups included both males and females. RESULTS: The results showed that 0.61% of the children with BCLP and 0.40% of those with UCLP had an intranasal tooth. The prevalence of an intranasal tooth for the whole group was 0.48%, and it appeared to be more common in females. PMID- 10830807 TI - Descriptive epidemiology of nonsyndromic cleft lip with or without cleft palate in Shanghai, China, from 1980 to 1989. AB - OBJECTIVE: The purpose of this study was to characterize nonsyndromic cleft lip with or without cleft palate (CL+/- CP) in an Asian population. DESIGN: Birth prevalence was assessed in a large birth series in Shanghai, China. A 1:3 sex-age hospital matched case-control design was used to assess the effects of parental ages and pregnancy history on risk of CL+/- CP. PARTICIPANTS: Records of live births from 1980 to 1989 in 22 hospitals in Shanghai, China, were reviewed, comprising 541,504 consecutive births, which is by far the largest such Chinese sample ever investigated. The case-control study included 528 (308 male, 220 female) nonsyndromic CL+/- CP cases and 1,563 (912 male, 651 female) controls. RESULTS: From 1980 to 1989, the overall birth prevalence was 1.2 per 1,000 live births with statistically significant seasonal variation (more CL+/- CP births in January to July). The overall male:female ratio was 1.40:1. For males, statistically significant associations were identified with maternal age for the most severe clefts (bilateral overall, and also bilateral CL+CP subgroup). For females, statistically significant association was shown for pregnancy age with birth order (overall and in most subgroups). CONCLUSIONS: The birth prevalence of CL+/- CP in this Asian population was similar to published Caucasian rates. The observed seasonal variation would be consistent with possible environmental factors. Significant associations with maternal age, pregnancy age, and birth order warrant additional study of pregnancy history in Asian CL+/- CP. PMID- 10830808 TI - Comparative study between children with and without cleft lip and cleft palate, part 1: cephalometric analysis. AB - OBJECTIVE: This study was conducted to compare craniofacial relationships, position, and curvature of the cervical spine between children with cleft lip and cleft palate who had been operated on and children without clefts. METHOD: This study was performed in 28 children with mixed dentition. They were divided into two groups. The study group included 14 children with unilateral operated cleft lip and cleft palate, ranging in age from 6 to 12 years, who clinically presented with a short upper lip, abnormal lip seal, and inhibition of sagittal development of the midface that was radiographically assessed. The control group included 14 children without clefts, ranging in age from 8 to 11 years. All of them had normal lip seal, nasal breathing, and a clinically normal body posture. DESIGN: A lateral craniocervical radiograph in a self-balanced natural head position in an erect posture, and without using a head holder, was taken for each child of both groups, with the mandible in maximum intercuspation and lips in habitual posture. The true vertical was marked on all the films. Specific angular and linear dimensions were used to assess the craniocervical relationships, as were the position of the cervical spine, its curvature, or both. RESULTS AND CONCLUSIONS: The study group presented a significant increase in the extension of the head on the neck, forward position of the cervical spine, and a decrease in the curvature of the cervical spine in comparison with the children without clefts. These results are more relevant considering that the study group also presented higher significant values of lower facial height than children without clefts. PMID- 10830809 TI - Comparative study between children with and without cleft lip and cleft palate, part 2: electromyographic analysis. AB - OBJECTIVE: This study was conducted to compare electromyographic (EMG) activity of superior orbicularis oris muscle between children with repaired cleft lip and cleft palate and children without clefts. METHODS: This study included 28 children with mixed dentition. They were divided into two groups. The study group included 14 children with repaired unilateral cleft lip and cleft palate, ranging in age from 6 to 12 years, who presented clinically with a short upper lip, abnormal lip seal, and inhibition of sagittal development of the midface as assessed radiographically. The control group included 14 children without clefts ranging in age from 8 to 11 years. All had normal lip seal, nasal breathing, and a clinically normal body posture. DESIGN: Bipolar surface electrodes were used for EMG recordings of resting level activity and during swallowing of saliva, speech, and chewing and swallowing of an apple. RESULTS AND CONCLUSIONS: A significantly higher level of activity at rest and during swallowing of saliva was observed in the cleft lip and cleft palate group. Similar activity during speech and chewing and swallowing of an apple was observed in both groups. The higher level of activity at rest and during swallowing of saliva in children with cleft lip and cleft palate seems to suggest that upon higher functional demands their activity increases less than in children without clefts. From a clinical point of view, if increased EMG activity at rest and during swallowing of saliva reflects increased force on the maxilla, then our findings may corroborate Bardach's findings (1990) that surgical treatment of cleft lip has an iatrogenic effect on facial growth, although the lack of significant correlation between the cephalometric data and EMG findings in the present study. PMID- 10830810 TI - Three-dimensional computer morphometry of the maxilla and face in infants with Pierre Robin sequence--a comparative study. AB - OBJECTIVE: To analyze the morphology of the maxillary crest in infants with Pierre Robin sequence using an anthropometric coordinate system and to compare the data with those of healthy infants. SETTING: The study was performed at a craniofacial center servicing a large geographic area. PARTICIPANTS: The study involved eight infants aged 1-28 days (average, 7 days) with an established diagnosis of Pierre Robin sequence and six healthy infants aged 1-43 days (average, 22 days). MAIN OUTCOME MEASURES: Physical models of the maxilla and face obtained by alginate replication were analyzed by computer morphometry yielding the three-dimensional topology of the maxillary crest. RESULTS: The maxillary crest of children with Pierre Robin sequence shows an increased inclination relative to the transverse plane (30 +/- 3.9 degrees) as compared with that of healthy infants (20 +/- 2.9 degrees). The maxillary crest of the patients is shortened in the sagittal direction by comparison with healthy controls. CONCLUSIONS: The increased inclination of the maxilla in infants with Pierre Robin sequence may aggravate the retroposition of the mandible and may thus be a pathogenetic factor contributing to the severe respiratory problems. PMID- 10830811 TI - Craniofacial comparisons in 22-month-old lip-operated children with unilateral complete cleft lip and palate and unilateral incomplete cleft lip. AB - OBJECTIVE: The objective of the study was to analyze the craniofacial morphology in infants with unilateral complete cleft lip and palate (UCCLP) in which the lip and the anterior part of the palate had been surgically closed at 2 months of age and to compare the morphology with that of a control group with unilateral incomplete cleft lip (UICL) in which the lip had also been surgically closed at 2 months of age. DESIGN: The sample consisted of a total of 108 cleft children all fulfilling the entry criteria, besides diagnosis, as follows: The child was of Danish origin; the age of the child was between 650 and 750 days (approximately 22 months) at the time of examination; the child was healthy except for its single cleft malformation; the surgical procedure in each group had been performed at about 2 months of age by the same surgeon. The surgical methods used were a Tennison procedure (UICL group) and a Tennison procedure supplemented by palatovomer plasty (UCCLP group). METHODS: The method of investigation was infant cephalometry in the lateral, frontal, and axial projections. Linear, angular, and area variables describing the craniofacial morphology were calculated and supplemented by mean plots from the cephalometric projections in the two groups. RESULTS AND CONCLUSIONS: Statistical analysis based on Student's t test showed that the facial morphology in the 22-month-old UCCLP group differed significantly from that of the UICL group. The most pronounced differences were found in the maxillary complex and the mandible. The deviations observed in the UCCLP group at 22 months of age were similar to those previously observed at 2 months of age. However, several of the dysmorphic traits had become less pronounced; some had remained the same; and a few had become worse with time. PMID- 10830812 TI - Orthodontic treatment for a patient with Pierre-Robin sequence complicated by juvenile periodontitis. AB - OBJECTIVE: To arrest the advancement of periodontitis. PATIENT: A 17-year-old boy diagnosed with Pierre-Robin sequence at birth exhibited localized juvenile periodontitis. Severe bone loss and mobile teeth were localized in the incisors and molars, which were irregularly positioned, possibly associated with a residual scar from palatoplasty for a soft palate cleft at an early age. CONCLUSION: Combined orthodontic-periodontic-prosthodontic treatment arrested the advancement of the periodontitis and established a stable occlusion. PMID- 10830813 TI - Long term outcome analysis of two treatment methods for cleft palate: combined levator retropositioning and pharyngeal flap versus double-opposing Z-plasty. PMID- 10830814 TI - The morbidity of harvesting bone from the iliac crest. PMID- 10830815 TI - Vasectomy reversal. PMID- 10830816 TI - Diagnosis and treatment of post-herniorrhaphy vas deferens obstruction. AB - Childhood inguinal herniorrhaphy (IH) is one of the most frequent causes of seminal tract obstruction. The incidence of vasal obstruction was found to be as high as 26.7% in subfertile patients with a history of childhood IH. The distal end of the vas deferens was found at the internal inguinal ring or in the pelvic cavity in 56.7% of cases, more than 3 cm of the vas deferens had been resected in 37.9% of cases, and sperm was found in vasal fluid in 45.5% of cases during corrective surgery. Microsurgical two-layer vasovasostomy resulted in the postoperative appearance of sperm in 39% of patients. In patients with postoperative azoospermia, a secondary epididymal obstruction caused by a long term vasal obstruction is a highly probable cause. Ipsilateral epididymovasostomy following successful inguinal vasovasostomy results in the postoperative appearance of sperm in the ejaculate in 100% of the patients and a subsequent natural pregnancy rate of 50%. The overall pregnancy rate among couples, following surgery in 18 patients, was 43%, excluding pregnancies achieved by in vitro fertilization or intracytoplasmic sperm injection. Microsurgical reanastomosis of the seminal tract resulted in high impregnation rates among partners of patients with seminal tract obstruction caused by childhood IH. After receiving sufficient information on each treatment modality, patients can choose their preferred treatment, either reanastomosis of the seminal tract or assisted reproductive technology using epididymal or testicular sperm. PMID- 10830817 TI - Microsurgical epididymovasostomy. AB - Epididymal obstruction is found in about 30% of obstructive azoospermic patients. Half of these obstructions are due to unknown etiologies, followed by obstruction due to epididymitis. The development of a specific, mucosa-to-mucosa anastomosis of the epididymal tubule to the vasal mucosa using a surgical microscopy has greatly improved the outcome of epididymovasostomy. The patency and natural pregnancy rates in this study were 79% and 38%, respectively. Side-to-end anastomosis also decreases the technical difficulty in comparison to the end-to end anastomosis procedure. An anastomosis at the epididymal caput generally results in a poorer outcome than at the corpus or cauda epididymis. PMID- 10830818 TI - Transurethral resection of the ejaculatory duct. AB - Complete bilateral ejaculatory duct obstruction has long been recognized as an uncommon, treatable form of male infertility. Partial ejaculatory duct obstruction reflects a disturbance of ejaculation where sperm quality is impaired during transit through the distal vas deferens and ejaculatory ducts. With the advent and increased use of high-resolution transrectal ultrasonography, abnormalities of the distal ejaculatory ducts related to infertility have been well documented. Although there are no pathognomonic findings associated with ejaculatory duct obstruction, several clinical findings are highly suggestive. In an infertile man with oligospermia or azoospermia with low ejaculate volume, normal secondary sexual characteristics, testes and hormonal profile and dilated seminal vesicles, midline cyst, or calcification on transrectal ultrasonography, ejaculatory duct obstruction is suggested. Of course, other causes of infertility may be concomitantly present and need to be searched for and treated as well. In selected cases, transurethral resection has resulted in marked improvement in semen parameters and pregnancies have been achieved. As is the case with all surgical procedures, proper patient selection and surgical experience are necessary to obtain optimal results. However, it appears that the treatments currently available for relief of ejaculatory obstruction are not optimally effective. Only approximately one half of treated patients will have an improvement in semen parameters and only about one quarter of treated patients will contribute to a pregnancy. What remains to be determined is how to manage the additional nearly 50% of patients who do not benefit from transurethral resection of ejaculatory obstruction. Based on my experience, I suggest that transrectal ultrasonography should be the first diagnostic procedure used when infertile men are suspected of having ejaculatory duct obstruction; however, vasography should still be considered for a more comprehensive diagnosis of ejaculatory duct obstruction. In patients showing atrophic seminal vesicles on transrectal ultrasonography and having a history of pulmonary tuberculosis, further study is not necessary and microscopic epididymal sperm aspiration is recommended for in vitro fertilization. Qualitative measurement of semen fructose may be helpful in the diagnosis of partial ejaculatory duct obstruction. Patients having midline cyst and being treated by transurethral resection are expected to have the best outcome. PMID- 10830819 TI - Microsurgical subinguinal varicocelectomy. PMID- 10830820 TI - Nerve-sparing retroperitoneal lymph node dissection: control mechanism, technique for nerve-sparing and reconstruction. PMID- 10830821 TI - One-day penile prosthesis surgery by penile block. PMID- 10830822 TI - Histology of the fetal prune belly syndrome with reference to the efficacy of prenatal decompression. AB - BACKGROUND: Deficient abdominal musculature, complex abnormalities of urinary tracts and bilateral abdominal cryptorchidism represent the basic characteristics of prune belly syndrome (PBS). Although prenatal diagnosis of PBS is rarely made, because of the wide variety of ultrasonographic images, reported cases have gradually increased. Once a fetus suspected of having PBS is found, it is sometimes difficult for the pediatric urologists to decide how to treat them. The histology of the kidney and urinary tracts in fetuses with PBS was reviewed in order to give suggestions on the management of prenatal cases. METHODS: Autopsy records of nine fetuses (5 males, 2 females and 2 undetermined) with characteristically distended and deficient abdominal wall were reviewed. Gestational age (GA) at detection ranged from 12 to 25 weeks and at delivery from 13 to 32 weeks. RESULTS: Renal histology in two fetuses showed earlier than normal disappearance of cortical nephrogenic zone replaced by cortical cysts and dysplastic structures. The nephrogenic zone was retained in five fetuses which were younger than GA 20 weeks. While the number of glomeruli along the medullary ray was normal for the age in three fetuses younger than GA 20 weeks, it was decreased in all others. Bladder histology was variable showing both increased musculature and defective or dysplastic muscles. There was a tendency for connective tissues in the bladder wall to increase in proportion to GA, The ureter revealed scarcity of muscle bundles among dense connective tissue. The urethra was atretic in eight fetuses. CONCLUSION: The clinical implication from the renal histology is that decompression of the urinary tract should be done before GA 20 weeks. However, the early fetal treatment appears to have no effect on the urodynamics in this disorder with deficient musculature. PMID- 10830823 TI - What is the optimal treatment for lower ureteral stones larger than 1 cm? AB - PURPOSE: To compare the treatment options for lower ureteral stones larger than 1 cm. METHODS: The records of 449 patients with lower ureteral calculi larger than 1 cm were reviewed retrospectively. Of these patients 342 (76.1%) were treated with extracorporeal shock wave lithotripsy (ESWL) (group 1), 66 (14.7%) with pneumatic lithotripsy (PL) (group 2) and 128 (28.5%) with ureterolithotomy (group 3). Eighty-seven (19.5%) patients underwent any of the two treatment modalities because of unsuccessful primary treatment. RESULTS: The overall stone-free rates were 32.4, 90.9 and 95.3% for ESWL, PL and ureterolithotomy, respectively. These values were 84.4% for primary PL and 96.7% for primary ureterolithotomy. The re treatment rate (46.4%) and secondary procedures were much more frequent in the ESWL group. There was no difference in the complication rates of the three groups. CONCLUSIONS: Pneumatic lithotripsy with ureteroscopy seems to be an appropriate treatment for larger ureteral stones. While ESWL can be tried as a first treatment option because of its noninvasive nature, lower success and higher re-treatment rates limit its usefulness. Ureterolithotomy is still a reasonable alternative for these large or unfragmented stones. PMID- 10830824 TI - Comparative study of pulsed dye laser and pneumatic lithotripters for transurethral ureterolithotripsy. AB - BACKGROUND: Clinical effectiveness and safety of the Swiss Lithoclast (Lithoclast) and the Candela MDL-2000 (MDL) in the treatment of lower ureteral stone were examined retrospectively. METHODS: Eighty-six stones from 66 patients and 26 stones from 20 patients were treated by Lithoclast and MDL, respectively. RESULTS: The stone-free rate on 3-month follow-up was 97% and 95% for the Lithoclast and MDL, respectively (no significant difference). The operation time was significantly shorter for the Lithoclast than for the MDL (90.2+/-50.2 vs 120.4+/-55.1 min; P<0.05). Postoperative analgesics were required significantly less frequently in Lithoclast (10/66 vs 11/20; P<0.01). Postoperative hospital stay was significantly shorter for Lithoclast (8.7+/-5.1 vs 12.1+/-4.2 days; P<0.01). CONCLUSIONS: Swiss Lithoclast is an effective and less invasive modality for endoscopic treatment of lower ureteral stones. PMID- 10830825 TI - Urothelial cytoprotective activity of Tamm-Horsfall protein isolated from the urine of healthy subjects and patients with interstitial cystitis. AB - PURPOSE: Tamm-Horsfall protein (THP) is a ubiquitous urinary protein with essentially no known function. We propose that THP is a cytoprotective agent that protects the urothelium from cationic species. To test this hypothesis we isolated THP from normal and interstitial cystitis urine to see if it could protect cultured cells from damage induced by the polyamine, protamine sulfate (PS). METHODS: Tamm-Horsfall protein was extracted from the urine of interstitial cystitis (IC) patients (N=28) and normal volunteers (N=5). Urothelial target cells (T24) were radiolabeled with 51Cr and then exposed to PS (0-1.0 mg/mL) for either 1.5 or 20 h. The resulting cytotoxicity data (dose-response curves) were then compared with the data obtained when PS was preincubated with 0-0.5 mg/mL of THP (IC vs normal), the semisynthetic polysaccharide, pentosan polysulfate (Elmiron), or human serum albumin. RESULTS: Toxicity of PS was significantly reduced by incubation with THP (or Elmiron) prior to evaluation by the chromium release assay, but not reduced by incubating with another protein, albumin. Tamm Horsfall protein from IC patients' urine was less protective than an equal quantity of THP from normal urine. CONCLUSIONS: These experiments suggest that THP has an important role in bladder mucosal defense mechanisms, protecting the bladder surface from injury. Inability of THP to prevent cytotoxic damage by urinary polyamine or other urinary toxins (cationic species) may be relevant in the etiology of interstitial cystitis, as putative urinary toxic components have been described in the urine of some patients. PMID- 10830826 TI - Primary carcinoid tumor in a horseshoe kidney. AB - A case of primary carcinoid tumor arising within a horseshoe kidney in a 51-year old woman is reported. The tumor was found incidentally by computed tomography (CT) during a check-up for a suspected gall bladder polyp. Histologic, immunohistochemical and electron microscopic analyses of this tumor revealed features typical of carcinoid tumor. Primary carcinoid tumor of the kidney is extremely rare and only 32 cases were previously reported, including five cases in horseshoe kidneys. None of these five cases in horseshoe kidneys demonstrated any evidence of local or distant metastases and all were alive at the time of reporting without evidence of disease after up to 3 years of follow up. The present case, even with accompanying lymph nodal metastasis, also has had no evidence of local recurrence or distant metastasis for 3 years post operation. Primary carcinoid tumor arising within horseshoe kidneys appear to be more benign than those within non-horseshoe kidneys. PMID- 10830827 TI - Interferon therapy for renal cell carcinoma in hemodialysis patients: report of two patients. AB - The method of interferon therapy has not been clarified in hemodialysis patients with renal cell carcinoma. Two hemodialysis patients with renal cell carcinoma were treated with natural interferon-alpha. The serum levels of interferon-alpha in both patients were measured after 24, 48, 72 and 96 h of injection. In case 1, the serum concentration of interferon-alpha after 24 h of injection at a dose of 3 x 10(6) reached a maximum and decreased gradually. In case 2, the serum concentration of interferon-alpha reached a maximum 24 h after injection at a dose of 3 x 10(6) and decreasing gradually after this. After an injection of interferon-alpha at a concentration of 6 x 10(6) 7 days later, the serum concentration of interferon-alpha reached a maximum 48 h after the injection and decreased gradually thereafter. The serum interferon-alpha concentrations of these cases were higher than normal renal function patients in other literature. It is necessary to modify the interval between injections for hemodialysis patients with renal cell carcinoma. PMID- 10830828 TI - Cystic renal cell carcinoma diagnosed by cystofiberscopy. AB - A 64-year-old man suffered from hypertensive syncope and was admitted to Branch Hospital, Faculty of Medicine, The University of Tokyo. On admission he had no symptoms; however, two renal cysts of about 5 cm in diameter were found in his left kidney. Cystofiberscopy with a thin optical fiberscope was performed in addition to puncture and cystography. Cytology revealed class I, but the wall surface of the lower cyst was uneven, fluid-filled, colloid-like and white-gray compared with that seen in the upper cyst. It was diagnosed as renal cell carcinoma (RCC) and left nephrectomy was performed (5 cm in diameter, RCC, papillary tubular, common, clear cell subtype, G2>G1, INF beta, pT2, pMx, pV0, ew(-)). PMID- 10830829 TI - Congenital vesicovaginal fistula. AB - Congenital vesicovaginal fistula is a very rare entity, the etiology of which has not been clearly elucidated because pathologic features have not been mentioned in previous reports. The case of a 4-year-old girl having incontinence resulting from a congenital vesicovaginal fistula joining with the left ectopic ureter from the hypoplastic kidney is described. This is thought to be the first presentation of congenital vesicovaginal fistula joining with ectopic ureter. A microscopic examination revealed the fistula consisting of transitional cell epithelium, suggesting an abnormal fusion of the ureteral bud and caudal end of the mullerian duct with the urogenital sinus. PMID- 10830830 TI - Cladistic analysis of myxozoan species with known alternating life-cycles. AB - The phylogenetic relationships of 15 myxozoan taxa with known alternating life cycles were investigated in order to provide insight into the puzzling matches between myxosporeans and actinosporeans of the myxozoan life-cycle data. Phylogenetic analyses were performed using two partitioned data-sets of life cycle stages, myxosporean stage from fish hosts versus actinosporean stage from annelid hosts, and a combined data-set of myxosporean and actinosporean stages. A cnidarian parasite of fish, Polypodium hydriforme Ussov, 1885, was used as the outgroup. The supraspecific level grouping in the conventional classification of actinosporeans was not supported in the analysis of the partitioned data from the actinosporean phase, which yielded two equally parsimonious trees. Analysis of the partitioned data from the myxosporean phase provided 24 equally parsimonious trees and did not support the current classification of myxosporeans. The analyses of the partitioned data of myxozoans by life-cycle stage revealed a lack of taxonomic congruence between the two life-stage partitions. Two equally parsimonious trees were obtained from analysis of the combined data. The suborder Variisporina of the Myxozoa was not supported by the total evidence trees, while the monophyly of the species of Myxobolus Butschli, 1882 and of the Myxidiidae were supported. The cladograms from the combined data revealed that these myxozoan species formed four major monophyletic groups. Among them, two were supported by the partitioned data of the actinosporean phase. The phylogenetic signals and the better resolution reflected by the trees of combined data suggest that the phylogenetic total evidence approach should be employed in future studies of the systematics of myxozoans. PMID- 10830831 TI - A new species of Neoheterocotyle Hargis, 1955 (Monogenea: Monocotylidae) from the gills of Pristis clavata Garman (Pristidae) from Darwin, Australia. AB - Neoheterocotyle darwinensis n. sp. is described from between the secondary gill lamellae of the dwarf sawfish Pristis clavata Garman (Pristidae) collected at the mouth of Buffalo Creek near Darwin, Northern Territory, Australia. This is only the second monocotylid species to be described from northern Australia. N. darwinensis is distinguished from the other seven valid species in the genus by the morphology of the hamuli, the dorsal haptoral accessory sclerites and the male copulatory organ. The similarities between N. darwinensis and Nonacotyle pristis Ogawa, 1991 from the gills of the freshwater sawfish Pristis microdon Latham collected in Papua New Guinea are discussed. PMID- 10830832 TI - Diplobulbus brayi n. sp. (Digenea: Opecoelidae) from Odax acroptilus (Perciformes: Odacidae) off Western Australia. AB - Diplobulbus brayi n. sp. is described from the small intestine of Odax acroptilus (Perciformes: Odacidae) from Point Peron in southern Western Australia. One of the five previously described species of Diplobulbus, D. vitellosus Bilqees, 1972 from Tetraodon lunaris (now Lagocephalus lunaris), is synonymised with Alloheterolebes indicus (Gupta, 1968) Hafeezullah, 1971, which is known from the same host in the Indian Ocean. D. brayi n. sp. can be distinguished from the remaining four species of Diplobulbus by the distinct interruption of the vitelline follicles at the ventral sucker, its more elongate formand its long post-testicular space. Diplobulbus is so far restricted to the Indo-Pacific region and appears to bestenoxenic to labroid fishes. This is the first record of a trematode from an odacid fish. PMID- 10830833 TI - A new macradeninine, Zebrasomatrema pichelinae n. g., n. sp. (Digenea: Lecithasteridae), from tangs (Perciformes: Acanthuridae) off the southern Great Barrier Reef, Queensland, Australia. AB - A new genus, Zebrasomatrema, with the type-species Z. pichelinae n. sp., is described from Zebrasoma veliferum at Heron and Lizard Islands, Queensland, Australia, and Z. scopas, Acanthurus lineatus and A. triostegus at Heron Island. The new genus shares with Acanthurotrema an anterior flange, 14 subglobular vitelline lobes, a divided seminal vesicle and a long aglandular duct joining the seminal vesicle and the short pars prostatica, but differs in its quadripartite seminal vesicle, single vitelline field, lack of a sinus-sac and excretory arms which unite in the forebody. This report constitutes the first record of a macradeninine lecithasterid in Australian waters. A key to the Macradenininae is presented. PMID- 10830834 TI - Amapacanthus amazonicus n. g., n. sp. (Acanthocephala: Diplosentidae: Allorhadinorhynchinae) from Arius passany and Anableps microleps (Pisces) at Maraca Island off northern Brazil. AB - Amapacanthus amazonicus n. g., n. sp. is described from the intestine of Arius passany (Valenciennes) and Anableps microleps Muller. The most important diagnostic features are: a small globular proboscis armed with 6 diagonal rows of 3 stout hooks; middle hooks conspicuously stouter and larger than anterior ones; terminal hooks as long as middle hooks but straighter and more slender; a double walled proboscis receptacle; a trunk bearing spines anteriorly; and two tubular cement glands in the males. Amapacanthus n. g. is differentiated from Allorhadinorhynchus, Golvanorhynchus and Slendrorhynchus, the other genera of the Allorhadinorhynchinae, by the presence of a globular proboscis armed with a small number (18) of hooks. A key to the species of the Allorhadinorhynchinae is presented. PMID- 10830835 TI - Systematic status of Laurotravassoxyuris bravoae Osorio-Sarabia, 1984 (Nematoda: Pharyngodonidae) [= Atractis bravoae (Osorio-Sarabia, 1984) n. comb.: Cosmocercidae]. AB - Re-examination of type-specimens of the nematode Laurotravassoxyuris bravoae Osorio-Sarabia, 1984, originating from the type-host, Cichlasoma istlanum Jordan et Snyder, from Mexico, has shown that these do not belong to Oxyuroidea Cobbold, 1864; in fact, the specimens included in the type-series of L. bravoae belong to two different families of the Cosmocercoidea Railliet, 1916: the Atractidae Railliet, 1917 and the Cosmocercidae Railliet, 1916. Both forms are described. Since the holotype of L. bravoae belongs to the former family, this species is transferred to Atractis Dujardin, 1845 as A. bravoae (Osorio-Sarabia, 1984) n. comb., whereas the female nematodes, belonging to the latter family, cannot be explicitly assigned to a genus or a species (although they might be conspecific with Raillietnema kritscheri Moravec, Salgado-Maldonado & Pineda-Lopez, 1993). A. bravoae is the first species of Atractis reported from fishes; its morphology differs distinctly from that in other congeners. PMID- 10830836 TI - Three little known acanthostomine digeneans from Uranoscopus scaber L. in the western Mediterranean. AB - Three species of acanthostomine digeneans, Anisocoelium capitellatum from the gall-bladder and Anisocladium fallax and A. gracilis from the intestine, are redescribed from the teleost Uranoscopus scaber in the western Mediterranean. The latter two species, which have in the past been confused and synonymised, are distinguished clearly on the basis of morphology, size and site. PMID- 10830837 TI - [The Cymothoidae (isopod parasites of marine fishes) of the coast of Tunisia: ecology and parasitological index]. AB - The study of the cymothoid isopod parasites on marine fishes from Tunisian localities has allowed us to distinguish eight species, seven of which have already been recorded and one, Mothocya epimerica which is new to the region. New hosts for Cerathotoa parallela and Nerocila orbignyi are reported. For each parasite species collected, the host fish, the parasitic specificity and the parasitological index are given. PMID- 10830838 TI - The distribution of monogenean parasites on aquatic vertebrates inhabiting Chinese inland waters. AB - The general distribution of monogenean parasites of aquatic vertebrates inhabiting Chinese inland waters is summarised. Five hundred and seventy-two out of a total of 581 species of monogeneans were discovered on fish, while only nine species were found on Amphibia and Reptilia. Most dactylogyrids and diplozoids parasitise cyprinids, while ancyrocephalids occur on silurids and cyprinids, and gyrodactylids are found mainly on cyprinids and cobitids. Analyses of host specificity and host-diversity suggests that the family Ancyrocephalidae should be divided into several families parallel with the Dactylogyridae. Of 12 subfamilies of the Cyprinidae, only the Gobiobotinae was found to be free of infection with monogeneans, and the genus Dactylogyrus has more species than any other monogenean genus associated with every cyprinid subfamily. PMID- 10830839 TI - Ancient metalloenzymes as possible markers in molecular archaeology. AB - The successful preparation of an active remnant of Cu,Zn-superoxide dismutase from mummified brain tissue stimulated the isolation of both biochemically and immunologically active alkaline Zn2Mg-phosphatase from antique bone samples of different archaeological sites and age. In particular, specimens from pharaonic Egypt being up to 4000 years of age were used. Gel filtration, ion exchange and affinity chromatographies were employed to optimise the preparation of the ancient enzyme. Compared to the specific activity of alkaline phosphatase from modern autopsy some 50% for a Ptolemaic and 10% for the Old Kingdom enzyme was detectable. The possibility of microbial contamination was checked by employing specific monoclonal antibodies directed against the human bone enzyme. Fortunately, ubiquitously present specified microorganisms on the respective ancient bones did not cross-react with these antibodies while the ancient metalloenzyme reacted with high specificity. Alkaline phosphatase mimicks could be excluded as in the presence of the inhibitors 1,10-phenanthroline and L homoarginine the enzyme activity was diminished. The presence of ortho-vanadate as a substrate analogon abolished the catalytic function of the enzyme. Likewise, heating to 100 degrees C and replacement of zinc(II) by cadmium(II) resulted in a dramatic loss of activity. In conclusion, alkaline phosphatase appears to be a useful marker enzyme in molecular archaeology. PMID- 10830840 TI - Metal-thiolate clusters in neuronal growth inhibitory factor (GIF). AB - Human neuronal growth inhibitory factor (GIF) is a metallothionein-like protein specific to the central nervous system, which has been linked to Alzheimer's disease. In this article a short overview of the biological and structural properties of native Cu4,Zn3-GIF are described. Moreover, metal-thiolate clusters formed in the synthetic beta-domain (residues 1-32) and the alpha-domain (residues 32-68) both with native CuI and ZnII, and as a spectroscopic probe also with Cd(II) are discussed. The cluster formation was followed by electronic absorption, circular dichroism (CD), magnetic circular dichroism (MCD) and 113Cd NMR spectroscopy and, in the special case of Cu(I) complexes, by luminescence spectroscopy at 77 K. These structural features are compared with those of recombinant Zn7- and 113Cd7-GIF. The structural studies suggest the existence of distinct MeII4S11 and MeII3S9 clusters located in the mutually interacting alpha- and beta-domains, respectively, of Cd7-GIF. In addition, evidence for a highly dynamic and flexible structure of this protein is presented. PMID- 10830841 TI - Circular dichroism, kinetic and mass spectrometric studies of copper(I) and mercury(II) binding to metallothionein. AB - The metalloprotein metallothionein (MT) is remarkable in its metal binding properties: for the mammalian protein, well-characterized species exist for metal to sulfur ratios of M7S20, M12S20, and M18S20, where M = Cd(II), Zn(II), Hg(II), Ag(I), Au(I), and Cu(I). Optical spectra in general, and circular dichroism (CD) and luminescence spectra in particular, provide rich detail of a complicated metal binding chemistry when metals are added directly to the metal-free or zinc containing protein. CD spectral data unambiguously identify key metal to protein stoichiometric ratios that result in well-defined structures. Electrospray ionization-mass spectrometry data are reported for reactions in which Hg(II) binds to apo-MT 2A as previously described from CD data. Emission spectra in the 450-750 nm region have been reported for metallothioneins containing Ag(I), Au(I), and Cu(I). The luminescence of Cu-MT can also be detected directly from mammalian and yeast cells. We report both steady-state and new dynamic data for titrations of Zn-MT with Cu(I). Analysis of kinetic data for the addition of the first two Cu(I) atoms to Zn-MT indicates a first-order mechanism over a concentration range of 5-50 microM. Three-dimensional modeling was carried out using the results of the CD and EXAFS studies, model calculations for Zn7-MT, Hg7 MT, and Cu12-MT are described. PMID- 10830842 TI - On the mechanism of action of thiamin enzymes in the presence of bivalent metal ions. AB - Results on the interactions between the bivalent metal ions Zn2+, Cd2+, Hg2+, Co2+, Ni2+ and 'active aldehyde' thiamin derivatives are reviewed. The techniques used in these studies include spectroscopic methods, i.e., IR-Raman, UV-Vis, multidimensional and multinuclear NMR in solution and in solid state, and X-ray crystal structure determinations. More recently, potentiometric studies on thiamin pyrophosphate and 2-(alpha-hydroxyethyl)thiamin in combination with NMR and EPR techniques were also undertaken. All these studies lead to useful conclusions on the mechanism of action of thiamin enzymes in the presence of bivalent metal ions. PMID- 10830843 TI - Anion- and pH-linked conformational transition in horseradish peroxidase. AB - In a previous study we have shown that bringing horseradish peroxidase to pH 3.0 induces a spectroscopic transition (G. Smulevich et al., Biochemistry 36 (1997) 640). We have extended the investigation on this pH-linked conformational change to different experimental conditions, such as (i) in phosphate alone, (ii) in HCl alone and (iii) in phosphate + NaCl. The data obtained allow a number of conclusions to be drawn, namely: (a) the exposure to pH 3.0 under all conditions brings about an alteration of the distal portion of the heme pocket, leading to the rapid formation of a hexa-coordinated species; (b) only in the presence of phosphate is the hexa-coordination followed by a slow cleavage (or severe weakening) of the proximal Fe-His bond, and (c) the rate of this second process is speeded up in the presence of Cl- ions. Such observations underline the presence of a communication pathway between the two opposite sides of the heme pocket, such that any alteration of the structural arrangement on one side of the heme cavity is transmitted to the other, inducing a corresponding conformational change. PMID- 10830844 TI - Covalently modified microperoxidases as heme-peptide models for peroxidases. AB - Microperoxidase-8 (MP8) and microperoxidase-9 (MP9) have been covalently modified by attachment of proline-containing residues to the amino terminal peptide chain in order to obtain new peroxidase model systems. The catalytic activities of these derivatives in the oxidation of p-cresol by hydrogen peroxide have been compared to that of MP8. The presence of steric hindrance above the heme reduces the formation rate of the catalytically active species, while the reactivity is increased when the amino group of a proline residue is close to the iron. The modification of the catalyst affects the rate of degradation processes undergone by the heme group during catalysis. A bulky aromatic group on the distal side decreases the stability of the complex because it reduces the mobility of a phenoxy radical species formed during catalysis, while the presence of proline residues increases the number of turnovers of the heme catalysts before degradation. The complex Pro2-MP8 obtained by addition of two proline residues to MP8 exhibits the best catalytic performance in terms of activity and chemical stability. PMID- 10830845 TI - Design and synthesis of new models for diiron biosites. AB - In order to mimic dinuclear active sites of some non-heme diiron proteins, ten new polydentate and potentially dinucleating ligands have been synthesized. Each ligand contains a carboxylate moiety designed to bridge two metal atoms. These central carboxylate moieties are derived from substituted benzoic acids that in turn are linked to terminal nitrogen or oxygen donors by spacers so that framework-type polydentate ligands similar to the polypeptide frames in diiron metallobiosites are formed. Reaction of these ligands with Fe(ClO4)3 x 9H2O leads to ferric mu-oxo-mu-carboxylato iron complexes of the general formulas [Fe2O(L)2(H2O)2](ClO4)2 and [Fe2O(L)(BzO)](ClO4)2 (L = ligand), containing one or two immobilized bridging carboxylates, respectively. While X-ray crystallography shows that some of these complexes are dimers or network polymers in the solid state, electrospray ionization mass spectrometry (ESMS) and spectroscopic data (UV-Vis, NMR, Mossbauer) indicate that they dissociate to monomeric Fe2O units in dilute CH3CN solutions. PMID- 10830846 TI - Metal complexes of salicylhydroxamic acid (H2Sha), anthranilic hydroxamic acid and benzohydroxamic acid. Crystal and molecular structure of [Cu(phen)2(Cl)]Cl x H2Sha, a model for a peroxidase-inhibitor complex. AB - Stability constants of iron(III), copper(II), nickel(II) and zinc(II) complexes of salicylhydroxamic acid (H2Sha), anthranilic hydroxamic acid (HAha) and benzohydroxamic acid (HBha) have been determined at 25.0 degrees C, I=0.2 mol dm( 3) KCl in aqueous solution. The complex stability order, iron(III) >> copper(II) > nickel(II) approximately = zinc(II) was observed whilst complexes of H2Sha were found to be more stable than those of the other two ligands. In the preparation of ternary metal ion complexes of these ligands and 1,10-phenanthroline (phen) the crystalline complex [Cu(phen)2(Cl)]Cl x H2Sha was obtained and its crystal structure determined. This complex is a model for hydroxamate-peroxidase inhibitor interactions. PMID- 10830847 TI - Enzyme-mimetic model compounds: conformational analysis and far-IR study of Cu(TAAB)2+. AB - Many enzymes occurring in nature like superoxide dismutase are systems rather too big to be accessible for vibrational and quantum chemical investigations. Thus, enzyme-mimetic model compounds consisting of a biological active metal centre surrounded by a macrocyclic ligand are used to shed light on binding properties of the active metal centre. Far- and mid-range IR spectroscopic investigations and a conformational analysis with the semi-empirical ZINDO/1 method of superoxide dismutase-mimetic complex Cu[TAAB]2+ are performed (TAAB = [b,f,j,n][1,5,9,13]tetra-aza-cyclohexadecine (tetra-anhydroamino benzaldehyde)). A distorted tetrahedral copper(II) centre with slightly twisted phenyl subunits is determined as the most stable conformation. Calculated mid- and far-IR spectra are in good agreement with the experimental data and confirm the proposed structure. A harmonic normal-coordinate analysis is used to assign the vibrational modes of the observed spectra. PMID- 10830848 TI - New mechanistic insights into the reactivity of the R2 protein of E. coli ribonucleotide reductase (RNR). AB - Further to a linear free-energy correlation of cross-reaction rate constants k12 for the reaction of eight organic radicals (OR), e.g. MV*+, from methyl viologen, with cytochrome c(III), we consider here similar studies for the reduction of the R2 protein of Escherichia coli ribonucleotide reductase, which has FeIII2 and Tyr* redox components. The same two techniques of pulse radiolysis and stopped flow were used. Cross-reaction rate constants (22 degrees C) at pH 7.0, I=0.100 M (NaCl), were determined for the reduction of active-R2 with the eight ORs, reduction potentials E0(1) from -0.446 to +0.194 V. Samples of active-R2 have an FeIII2 met-R2 component, which in the present studies was close to 40%. Concurrent reactions have to be taken into account for the five most reactive ORs, corresponding to reduction of the FeIII2 of met-R2 and then of active-R2. Separate experiments on met-R2 reproduced the first of these rate constants, which on average is approximately 66% larger than the second rate constant. A single Marcus free-energy plot of log k12-0.5 log10f versus -E0(1)/0.059 describes all the data and the slope of 0.54 is in satisfactory agreement with the theoretical value of 0.50. Such behaviour is unexpected since the Tyr* is a much stronger oxidant (E0 approximately 1.0 V versus NHE) as compared to FeIII2 (E0 close to zero). X-ray structures of the met- and red-R2 states have indicated that electroneutrality of the approximately 10 A buried active site is maintained. Proton transfer is therefore proposed as a rapid sequel to electron transfer. Other reactions considered are the much slower conventional time-range reductions of active-R2 with hydrazine and dithionite. For these reactions one and/or two-equivalent changes are possible. With both reductants, met-R2 reacts about four-fold faster than active-R2, and as with the ORs the less strongly oxidising FeIII2 component is reduced before the Tyr*. PMID- 10830849 TI - Synthesis and reactivity studies of model complexes for molybdopterin-dependent enzymes. AB - The molybdenum cofactor (Moco)-containing enzymes are divided into three classes that are named after prototypical members of each family, viz. sulfite oxidase, DMSO reductase and xanthine oxidase. Functional or structural models have been prepared for these three prototypical enzymes: (i) The complex [MoO2(mnt)2]2- (mnt2- = 1,2-dicyanoethylenedithiolate) has been found to be able to oxidize hydrogen sulfite to HSO4- and is thus a functional model of sulfite oxidase. Kinetic and computational studies indicate that the reaction proceeds via attack of the substrate at one of the oxo ligands of the complex, rather than at the metal. (ii) The coordination geometries of the mono-oxo [Mo(VI)(O-Ser)(S2)2] entity (S2 = dithiolene moiety of molybdopterin) found in the crystal structure of R. sphaeroides DMSO reductase and the corresponding des-oxo Mo(IV) unit have been reproduced in the complexes [M(VI)O(OSiR3)(bdt)2] and [M(VI)O(OSiR3)(bdt)2] (M = Mo,W; bdt = benzene dithiolate). (iii) A facile route has been developed for the preparation of complexes containing a cis-Mo(VI)OS molybdenum oxo, sulfido moiety similar to that detected in the oxidized form of xanthine oxidase. PMID- 10830850 TI - Micropattern immobilization of polysaccharide. AB - Two types of polysaccharides, sulfated hyaluronic acid and heparin, were pattern immobilized on a poly(ethylene terephthalate) and polystyrene film, respectively, in a specific pattern by photolithography. Sulfated hyaluronic acid was prepared from hylaronic acid by the treatment of sulfur trioxide/pyridine complex. Heparin was purchased and used without further treatment. The polysaccharides were coupled with azidoaniline. The derivatized polysaccharides were cast on a film from aqueous solution. After drying, the film was photo-irradiated in the presence or absence of a photomask. The micropatterning was confirmed by staining with a cationic dye. Platelet adhesion was reduced on the sulfated hyaluronic acid-immobilized areas. The immobilized sulfated hyaluronic acid significantly reduced thrombus formation. On the other hand, cells were cultured on the heparin immobilized film. In the presence of fibroblast growth factor (FGF), the growth of mouse fibroblast STO cells was enhanced only on the heparin-immobilized regions. This result indicated that micropattern-immobilized heparin activated FGF for cell growth activity. PMID- 10830851 TI - Electron transfer properties of iron-sulfur proteins. AB - The details of most electron transfer reactions involving iron-sulfur proteins have remained undisclosed because of the lack of experimental methods suitable to measure precisely the relevant rates. Nuclear magnetic resonance (NMR) provides a powerful means to overcome these problems, at least with selected proteins. A combination of NMR studies and site-directed mutagenesis experiments has been instrumental in defining both the site of interaction and the main trends of the intracomplex electron transfer in the case of rubredoxin electron self-exchange. Analysis of the NMR data obtained for mixtures of different redox levels of several 2[4Fe-4S] ferredoxins provided both first-order, for intramolecular, and second-order, for intermolecular, rate constants. Their dependence as a function of structural changes gave insight into the mechanism of electron transfer in this type of protein. Contrary to some expectations, the high-spin [4Fe-4Se]+ clusters assembled in isopotential ferredoxins do not change the intramolecular electron transfer rate as compared to low-spin [4Fe-4S]+ homologs. In combination with activity measurements, the kinetic data have been used to model the electron transfer competent complexes between Clostridium pasteurianum ferredoxin and the main enzymes acting as redox partners in vivo. PMID- 10830852 TI - Design and synthesis of new biomimetic materials. AB - In this paper, it is reported that the histidine-silane derivative Boc-His(Boc) CONH-(CH2)3Si(OEt)3 can be polymerized via the sol-gel method or can be grafted on a silica surface. The obtained organosilicas bear histidine molecules covalently bonded on the inorganic matrix. Their Cu(II) complexes have been evaluated as oxidation catalysts for the conversion of 3,5-di-tert-butylcatechol (DTBC) to 3,5-di-tert-butylquinone (DTBQ) in the presence of dioxygen. PMID- 10830853 TI - Polymer-based composite hip prostheses. AB - A composite hip prosthesis (CHP) made from poly(ether-imide) reinforced with carbon and glass fibres was manufactured and characterized. The main objective of the study was to evaluate the effect of fibre organization on the mechanical properties of the composite femoral implant and compare with the bone. A stacking sequence of drop-off plies of carbon/glass fibres reinforcing poly(ether-imide) (PEI) constitutes a symmetrical and balanced CHP. The hip was manufactured according to the finite element modelling (FEM) design and using the compression moulding and water-jet technologies. The measured stress-strain data according to tensile, flexural and torsional tests showed agreement with the numerical calculation. Young's modulus and the strength in tension are uniform along the stem axis (40 GPa and 600 MPa, respectively) while the elastic modulus in bending varies from 10 to 60 GPa in the tip-head direction. The composite stem showed a linear load-displacement relation up to 4500 N without breaking. Mechanical behaviour of the CHP is compared to that of a canine femur. Comparison with metal prostheses has also been undertaken. CHPs control stress-strain distributions, and hence the mechanical signals to bone, through a material-structure design. PMID- 10830854 TI - Direct micellar systems as a tool to improve the efficiency of aromatic substrate conversion for fine chemicals production. AB - Whole-cell bioconversion of naphthalene to (+)-cis-(1R,2S)-dihydroxy-1,2 dihydronaphthalene by Escherichia coli JM109(pPS1778) recombinant strain, carrying naphthalene dioxygenase and regulatory genes cloned from Pseudomonas fluorescens N3, in direct micellar systems is optimized as an example of fine chemicals bioproduction from scarcely water-soluble substrates. The oxygen insertion into the aromatic substrate, which stops at the enantiomerically pure cis dihydroxylated product, is performed in direct microemulsion systems, where a non-ionic surfactant stabilizes naphthalene containing oil droplets in an aqueous medium. These media provide an increased substrate solubility so that a homogeneous reaction can be carried out, while not affecting bacteria viability and performances. The influence of the chemical nature of the oil is investigated. The phase behavior of the direct microemulsion system was monitored for three different oils as a function their volume fraction and characterized through light scattering. The addition of isopropyl palmitate, oleic acid, or glyceryl trioleate, 0.6-1.2% v/v to the micellar systems, led to an increase of the substrate concentration in the solution and particularly its bioavailability, allowing faster catalytic conversions. All these systems resulted in being suitable for catalytic conversions of aromatic compounds. Although the nature of the oil does have a deep effect on the phase behavior of the micellar systems, in the present investigation no differences in the yields and in the rates of product formation of the enzymatic system were observed on changing the oil, thus showing that in this case the substrate concentration or bioavailability is not the rate-limiting step. PMID- 10830855 TI - Nanoscale iron(III) oxyhydroxy aggregates formed in the presence of functional water-soluble polymers: models for iron(III) biomineralisation processes. AB - Superparamagnetic clusters of iron(III) oxyhydroxide in the form of poorly crystalline ferrihydrite (formally, 5Fe2O3 x 9H2O) have been synthesised in the presence of the polymers polyvinyl alcohol (PVA), polyacrylic acid (PAA) and alginic acid. The solutions have been characterised by viscosity studies and the resultant arrays isolated from these solutions have been imaged under an electron microscope and their magnetic properties determined by 57Fe-Mossbauer spectroscopic studies at 293 and 77 K and magnetisation measurements at 293 and 5 K. The magnetic data show that the iron(III) oxyhydroxy particles are superparamagnetic. All preparations show hysteretic behaviour with coercive fields being approximately half or less than half of that of ferrihydrite (3.4 kOe) and values of magnetic moment per iron particle less than that of ferrihydrite. Nanoscale aggregates (2-4 nm) are formed in the presence of PVA and PAA while, with alginic acid, extended branch-like structures are observed, their formation being facilitated by the comparatively rigid polysaccharide chain, a process related to iron biomineralisation in diverse biological systems. PMID- 10830856 TI - Immobilisation of sulphated hyaluronan for improved biocompatibility. AB - Hyaluronan (Hyal) was modified by the insertion of sulphate to hydroxyl groups. A series of heparin-like compounds with controllable properties was obtained. The physicochemical and biological behaviours of all these sulphated hyaluronan acids (HyalSx) and their complexes with heavy metal ions (Cu2+ and Zn2+) were investigated. HyalS, derivatives showed a good anticoagulant activity and low platelet aggregation which increased with increasing degree of sulphation. Moreover HyalSx and their Cu2+ complexes were demonstrated to favour the growth of human endothelial cells. However, the utilisation of HyalSx as a material is hindered by its high solubility in physiological solution. Our approach to improve its stability was directed to the synthesis of new HyalSx-based hydrogels and on the preparation of new biocompatible polymeric surfaces obtained through covalent photoimmobilisation of HyalSx. The reaction of primary ovine chondrocytes and B10D2 endothelial cells was studied on both matrices in terms of cell number, F-actin and CD44 receptor immunostaining. Analysis of cell movement showed that the cells respond to HyalSx showing good adhesion and spreading. These results suggest that HyalSx containing materials could be used as biomaterials to aid cartilage repair and vessel endothelisation. PMID- 10830857 TI - Thermodynamic, kinetic and structural studies on the ternary palladium(II) complexes of thioether ligands. AB - Potentiometric, calorimetric, NMR and stopped-flow kinetic studies were performed on the palladium(II) complexes of thioether and/or nitrogen donor ligands. The ternary systems always contained a tridentate ligand (dien, terpy and dianions of dipeptides, GlyGly, GlyAla and GlyMet) and a monodentate thioether (AcMet). The stability constants of thioether complexes were obtained by indirect potentiometric measurements using uridine as a competitive ligand. The thermodynamic parameters revealed that selectivity of palladium(II) for thioether binding can be significantly influenced by the other donor atoms around the metal ion. [Pd(terpy)]2+ and [Pd(GlyMet)] had the lowest affinity for thioether binding and it was explained by steric and electronic effects. Ternary complexes of nitrogen donors have higher thermodynamic stability constants than the thioether complexes, but rate constants of the substitution reactions revealed that formation of thioether complexes is the faster reaction. As a consequence, the thermodynamic equilibrium state of a multicomponent system is characterized by the coordination of N-donors, which are formed via the existence of thioether bonded intermediates. PMID- 10830858 TI - Synthesis, characterization and in vitro antitumor activity of di- and triorganotin derivatives of polyoxa- and biologically relevant carboxylic acids. AB - An overview of the development of anti-tumor organotin derivatives, sometimes as active in vitro as doxorubicin, is presented and discussed. Solubility in water is an important issue, dominating the in vivo testing of compounds with promising in vitro properties. Several water-soluble organotin compounds gave the best in vitro activities. Novel, useful organotin anti-tumor compounds should be designed toward improved water solubility. PMID- 10830859 TI - Design of copper-based anti-inflammatory drugs. AB - It has been shown that the inflammation associated with rheumatoid arthritis can be reduced using copper complexes. In order to improve the bioavailability of copper and hence efficacy of these complexes we have synthesized three different series of ligands, each having different characteristics. Thermodynamic results for copper(II) complexes for these polyamino, diaminodiamido and triaminodiamido ligands are presented. The polyamino ligands form the most stable complexes in vivo but tissue distribution studies in mice show that [Cu(3,6,9,12 tetraazatetradecanedioate)] is excreted rapidly, unchanged in the urine. The diamino ligand complexes are much less stable than their polyamino analogues and animal studies using [Cu(N,N'-bis[2-(dimethylamino)ethyl]-ethanediamide)H2] indicate that the complex dissociates in vivo and is excreted slowly via the liver. The triaminodiamido copper(II) complexes are approximately 2 log units more stable than their diamino analogues. Computer simulation calculations indicate that these complexes are also likely to dissociate in plasma. Measured partition coefficients, however, suggest the possibility of dermal absorption. PMID- 10830860 TI - Transition metal complexes of diclofenac with potentially interesting anti inflammatory activity. AB - An overview is given of the results of metal ion-diclofenac interactions. Several complexes have been synthesized at the University of Ioannina. Binuclear complexes, [Cu(L)2(H2O)]2 x 2H2O and [CuL2(S)]2 where S is H2O, EtOH, DMSO, (CH3)2CO and DMF, and mononuclear complexes, [MnL2(H2O)], [FeL2(H2O)2], [CoL2(H2O)2] x 0.5H2O, [CoL2(H2O)], [NiL2(H2O)2] x 2H2O, [NiL2] and [PdL2] x 2H2O, have been characterized by spectroscopy, X-ray crystallography and electrochemical studies. The catalytic activity of these complexes was correlated to the reduction potential. Some of the complexes of diclofenac exhibit very promising anti-inflammatory activity and act as antioxidant compounds, a property that is absent from diclofenac. PMID- 10830861 TI - Synthetic analogs for oxovanadium(IV/V)-glutathione interaction: an NMR, EPR, synthetic and structural study of oxovanadium(IV/V) compounds with sulfhydryl containing pseudopeptides and dipeptides. AB - The reaction of [VO(CH3COO)2(phen)] (phen = 1,10-phenanthroline) with the sulfhydryl-containing pseudopeptides (scp), N-(2-mercaptopropionyl)glycine (H3mpg), N-(2-mercaptopropionyl)cysteine (H4m2pc), N-(3 mercaptopropionyl)cysteine (H4m3pc) and the dipeptides glycylglycine (H2glygly) and glycyl-L-alanine (H2glyala), in the presence of triethylamine, results in the formation of the compounds Et3NH[VO(mpg)(phen)] (1), (Et3NH)2[VO(m2pc)] (4), [(Et3NH)2[VO(m3pc) (5), [VO(glygly)(phen)] x 2CH3OH (2 x 2CH3OH) and [VO(glyala)(phen)] x CH3OH (3 x CH3OH). Evidence for the molecular connectivity in 2 x CH3OH was established by X-ray crystallography, showing the vanadium(IV) atom ligated to a tridentate glygly2- ligand at the N(amine), N(peptide) and O(carboxylato) atoms. Combination of the correlation plot of the EPR parameters gz versus Az, together with the additivity relationship supported the prediction of the equatorial donor atom sets of the V(IV)O2+ center at various pH values for the V(IV)O2+-glutathione system considered in this study. Model NMR studies (interaction of vanadium(V) with the scp H3mpg) showed that there is a possibility of vanadium(V) ligation to glutathione. PMID- 10830862 TI - Rates of platination of -AG- and -GA- containing double-stranded oligonucleotides: effect of chloride concentration. AB - The kinetics of the reactions between 15N-labelled cisplatin and 14-base pair duplex oligonucleotides with either 5'-AG-3' or 5'-GA-3' groupings as the principal platination site are examined in the presence of 60-80 mM chloride by [1H,15N]HSQC 2D NMR spectroscopy. The presence of chloride at these concentrations results in a five-fold decrease in the rate of hydrolysis of cisplatin to cis-[PtCl(NH3)2(OH2)]+ and a two- to twenty-fold decrease in the rate of monofunctional adduct formation. The effects on the rate of closure from monofunctional to bifunctional adducts are less well established but some of these rates appear not to be significantly reduced by the presence of added chloride. The results provide a caution that the use of chloride to quench platination reactions may not be fully effective. PMID- 10830863 TI - Antimetastatic properties and DNA interactions of the novel class of dimeric Ru(III) compounds Na2[[trans-RuCl4(Me2SO)]2(mu-L)] (L = ditopic, non-chelating aromatic N-ligand). A preliminary investigation. AB - A novel class of dianionic Ru(III) dimers of formula Na2[[trans-RuCl4(Me2SO)]2(mu L)], with L = pyrazine (pyz, 1), pyrimidine (pym, 2), 4,4'-bipyridine (bipy, 3), and 1,2-bis(4-pyridine) ethane (etbipy, 4), was developed by us with the specific aim of assessing their antitumor properties. The dimers are in fact structurally related to the antimetastatic mononuclear compound (ImH) [trans-RuCl4(Me2SO)(Im)] (NAMI-A, Im = imidazole). Preliminary results concerning the antineoplastic activity of 1-4 against the murine MCa carcinoma model, a tumor which spontaneously metastasizes in the lungs, are reported. Similarly to what is normally observed with NAMI-A, the treatment with the dimeric complexes was scarcely effective against the growth of the primary tumor. However, dimers 1, 2, and 4 reduced very effectively the number and, in particular, the weight of lung metastases (to about 5% with respect to controls); in particular, Na2[[trans RuCl4(Me2SO)]2(mu-etbipy)] (4) was as effective as NAMI-A in reducing the spontaneous metastases at a dosage which, in terms of moles of ruthenium, is about 3.5 times lower compared to that normally used for NAMI-A. Furthermore, in vitro tests showed that dimers 1-4 are capable of forming interstrand cross-links with linearized plasmidic DNA in a time-dependent manner. All the dimeric species are more active in inducing cross-links compared to NAMI-A, and the dimer bridged by the etbipy ligand (4) is the most effective among those tested. PMID- 10830864 TI - Gas-phase metal ion (Li+, Na+, Cu+) affinities of glycine and alanine. AB - The gas-phase metal affinities of glycine and alanine for Li+, Na+ and Cu+ ions have been determined theoretically employing the hybrid B3LYP exchange correlation functional and using extended basis sets. All computations indicate that the metal ion affinity (MIA) decreases on going from Cu+ to Li+ and Na+ for both the considered amino acids. The absolute MIA values are close to the experimental counterparts with the exception of lithium for which a deviation of about 7 kcal/mol at the B3LYP level is obtained. The optimized structures indicate that Li+, Na+ and Cu+ prefer a bidentate coordination, bonding with both nitrogen and oxygen atoms of amino acids. PMID- 10830865 TI - Copper transport and its defect in Wilson disease: characterization of the copper binding domain of Wilson disease ATPase. AB - Copper is an essential trace element which forms an integral component of many enzymes. While trace amounts of copper are needed to sustain life, excess copper is extremely toxic. An attempt is made here to present the current understanding of the normal transport of copper in relation to the absorption, intracellular transport and toxicity. Wilson disease is a genetic disorder of copper transport resulting in the accumulation of copper in organs such as liver and brain which leads to progressive hepatic and neurological damage. The gene responsible for Wilson disease (ATP7B) is predicted to encode a putative copper-transporting P type ATPase. An important feature of this ATPase is the presence of a large N terminal domain that contains six repeats of a copper-binding motif which is thought to be responsible for binding this metal prior to its transport across the membrane. We have cloned, expressed and purified the N-terminal domain (approximately 70 kD) of Wilson disease ATPase. Metal-binding properties of the domain showed the protein to bind several metals besides copper; however, copper has a higher affinity for the domain. The copper is bound to the domain in Cu(I) form with a copper: protein ratio of 6.5:1. X-ray absorption studies strongly suggest Cu(I) atoms are ligated to cysteine residues. Circular dichroism spectral analyses suggest both secondary and tertiary structural changes upon copper binding to the domain. Copper-binding studies suggest some degree of cooperativity in binding of copper. These studies as well as detailed structural information of the copper-binding domain will be crucial in determining the specific role played by the copper-transporting ATPase in the homeostatic control of copper in the body and how the transport of copper is interrupted by mutations in the ATPase gene. PMID- 10830866 TI - Arsenite, arsenate and vanadate affect human erythrocyte membrane. AB - Effects of arsenite, arsenate and vanadate on human erythrocyte membrane have been assessed according to their routes passing through the membrane, their binding modes to the membrane and their influences on membrane proteins and lipids. The uptake of arsenate (1.0 mM) by cells approached a limit with intracellular arsenic of about 0.2 mM in 5 h, and was strongly inhibited (approximately 95%) by 4,4'-diisothiocyano-2,2'-disulfonic stilbene (DIDS), indicating that arsenate, similar to vanadate, passed across the membrane through the anion exchange protein, band 3. Arsenite (1.0 mM) influx reached a maximum of about 0.4 mM in 30 min, and was not inhibited by DIDS. The transformed species of arsenite bound to the membrane from cytosol. In contrast, arsenate bound rapidly from the outside, followed by releasing and re-binding. The binding to the membrane via sulfhydryl was indicated by the decrease of the sulfhydryl level of membrane proteins. Polyacrylamide gel electrophoresis in sodium dodecyl sulfate (SDS-PAGE) analysis revealed that the proteins, bands 1-3, were among the targets of arsenite, arsenate and vanadate. Their binding to the membrane also induced changes in the fluidity of membrane lipids and in the negative charge density in the outer surface of the membrane. PMID- 10830867 TI - Some factors affecting metal ion-monohydroxamate interactions in aqueous solution. AB - The chelating properties exhibited by a series of monohydroxamic acids (propanohydroxamic acid (Pha), hexanohydroxamic acid (Hha), benzohydroxamic acid (Bha), N-methyl-acetohydroxamic acid (MAha), N-phenyl-acetohydroxamic acid (PhAha) and 2-hydroxypyridine-N-oxide (PYRha)) towards copper(II), nickel(II), zinc(II), calcium(II), magnesium(II) and aluminium(III) ions were studied by pH metric, spectrophotometric and, in one case, by 27Al NMR methods. The results were compared with the corresponding data for metal ion-acetohydroxamate (Aha) and metal ion-desferrioxamine B (DFB) complexes. Changes of the substituents either on the carbon or on the nitrogen of the hydroxamate moiety caused a measurable effect on the chelate stability only in the case of aluminium(III) complexes. The aromatic derivative, PYRha, formed significantly more stable complexes than expected on the basis of the ligand basicity. The higher complex forming ability of DFB compared to monohydroxamic acids diminishes in the case of the largest calcium(II) ion. PMID- 10830868 TI - Molecular models in nickel carcinogenesis. AB - Nickel compounds are known human carcinogens, but the exact molecular mechanisms of nickel carcinogenesis are not known. Due to their abundance, histones are likely targets for Ni(II) ions among nuclear macromolecules. This paper reviews our recent studies of peptide and protein models of Ni(II) binding to histones. The results allowed us to propose several mechanisms of Ni(II)-inflicted damage, including nucleobase oxidation and sequence-specific histone hydrolysis. Quantitative estimations of Ni(II) speciation, based on these studies, support the likelihood of Ni(II) binding to histones in vivo, and the protective role of high levels of glutathione. These calculations indicate the importance of histidine in the intracellular Ni(II) speciation. PMID- 10830869 TI - Engineering outer-membrane proteins in Pseudomonas putida for enhanced heavy metal bioadsorption. AB - Metallothioneins (MTs) are small, cysteine-rich proteins with a strong metal binding capacity that are ubiquitous in the animal kingdom. Recombinant expression of MT fused to outer-membrane components of gram-negative bacteria may provide new methods to treat heavy-metal pollution in industrial sewage. In this work, we have engineered Pseudomonas putida, a per se highly robust microorganism able to grow in highly contaminated habitats in order to further increase its metal-chelating ability. We report the expression of a hybrid protein between mouse MT and the beta domain of the IgA protease of Neisseria in the outer membrane of Pseudomonas cells. The metal-binding capacity of such cells was increased three-fold. The autotranslocating capacity of the beta domain of the IgA protease of Neisseria, as well as the correct anchoring of the transported protein into the outer membrane, have been demonstrated for the first time in a member of the Pseudomonas genus. PMID- 10830870 TI - Bacterial metal-resistance proteins and their use in biosensors for the detection of bioavailable heavy metals. AB - We have expressed and purified metal-resistance and metal regulatory proteins from the bacterial determinants of resistance to heavy metals and utilised these in the development of biosensors for heavy metals. Both the metallothionein from the cyanobacterium Synechococcus PCC 7942 and the MerR regulatory protein from transposon Tn501 allow the detection of non-specific metal binding down to 10( 15) M concentrations of Hg(II), Cu(II), Zn(II) and Cd(II) in pure solution. Differential effects of the metals can be detected at both low and high concentrations, and the shape of the capacitance curves may reflect biologically relevant responses of the proteins to metals. Further work is required to establish the relationship between the detected binding of metal and the biological response of the protein, or to provide biosensors of use in the natural environment. PMID- 10830871 TI - Inhibition of antimutagenic enzymes, 8-oxo-dGTPases, by carcinogenic metals. Recent developments. AB - Nickel, cadmium, cobalt, and copper are carcinogenic to humans and/or animals, but the underlying mechanisms are poorly understood. Our studies have been focused on one such mechanism involving mediation by the metals of promutagenic oxidative damage to DNA bases. The damage may be inflicted directly in DNA or in the deoxynucleotide pool, from which the damaged bases are incorporated into DNA. Such incorporation is prevented in cells by 8-oxo-2'-deoxyguanosine 5' triphosphate pyrophosphatases (8-oxo-dGTPases). Thus, inhibition of these enzymes should enhance carcinogenesis. We have studied effects of Cd(II), Cu(II), Co(II), and Ni(II) on the activity of isolated bacterial and human 8-oxo-dGTPases. Cd(II) and Cu(II) were strongly inhibitory, while Ni(II) and Co(II) were much less suppressive. After developing an assay for 8-oxo-dGTPase activity, we confirmed the inhibition by Cd(II) in cultured cells and in the rat testis, the target organ for cadmium carcinogenesis. 8-Oxo-dGTPase inhibition was accompanied by an increase in the 8-oxo-dG level in testicular DNA. PMID- 10830872 TI - A discussion of the chemistry of oxidative and nitrosative stress in cytotoxicity. AB - Nitric oxide (NO) has been shown to be a key bioregulatory agent in a wide variety of biological processes, yet cytotoxic properties have been reported as well. This dichotomy has raised the question of how this potentially toxic species can be involved in so many fundamental physiological processes. We have investigated the effects of NO on a variety of toxic agents and correlated how its chemistry might pertain to the observed biology. The results generate a scheme termed the chemical biology of NO in which the pertinent reactions can be categorized into direct and indirect effects. The former involves the direct reaction of NO with its biological targets generally at low fluxes of NO. Indirect effects are reactions mediated by reactive nitrogen oxide species, such as those generated from the NO/O2 and NO/O2- reactions, which can lead to cellular damage via nitrosation or oxidation of biological components. This report discusses several examples of cytotoxicity involved with these stresses. PMID- 10830873 TI - Cadmium carcinogenesis in review. AB - Cadmium is an inorganic toxicant of great environmental and occupational concern which was classified as a human carcinogen in 1993. Occupational cadmium exposure is associated with lung cancer in humans. Cadmium exposure has also, on occasion, been linked to human prostate cancer. The epidemiological data linking cadmium and pulmonary cancer are much stronger than for prostatic cancer. Other target sites for cadmium carcinogenesis in humans (liver, kidney, stomach) are considered equivocal. In rodents, cadmium causes tumors at several sites and by various routes. Cadmium inhalation in rats results in pulmonary adenocarcinomas, supporting a role in human lung cancer. Prostate tumors and preneoplastic proliferative lesions can be induced in rats after cadmium ingestion or injection. Prostatic carcinogenesis in rats occurs only at cadmium doses below those that induce chronic degeneration and dysfunction of the testes, a well known effect of cadmium, confirming the androgen dependency of prostate tumors. Other targets of cadmium in rodents include the testes, adrenals, injection sites, and hematopoietic system. Various treatments can modify cadmium carcinogenesis including supplemental zinc, which prevents cadmium-induced injection site and testicular tumors while facilitating prostatic tumors. Cadmium is poorly mutagenic and probably acts through indirect mechanisms, although the precise mechanisms remain unknown. PMID- 10830874 TI - Metal ion-carbonyl oxygen recognition in complexes of acetyl phosphate. AB - Studies on acetyl phosphate (AcP2-), one of the so-called 'energy-rich' mixed acid anhydrides, are summarized. Based on stability constants determined by potentiometric pH titrations in aqueous solution, it is shown that the M(AcP) complexes of Ca2+, Mg2+, Mn2+, Cu2+, and Zn2+ are more stable than is expected from the basicity of the phosphate group of AcP2-. This observed stability increase is attributed to an additional interaction of the already phosphate coordinated metal ion (M2+) with the carbonyl oxygen of the anhydride unit. These conclusions are corroborated by the properties of the complexes of the hydrolysis stable acetonylphosphonate (AnP2-). The formation degrees of the various six membered chelates occurring in the M(AcP) and M(AnP) systems are presented and evidence is given that these chelates persist in mixed ligand complexes and that their formation degree is promoted by a low solvent polarity. The biological relevance of these results regarding carbonyl oxygen-metal ion interactions is briefly indicated. PMID- 10830875 TI - Controlling gene expression by zinc(II)-macrocyclic tetraamine complexes. AB - The zinc(II) complexes of 12-membered macrocyclic tetraamines (1,4,7,10 tetraazacyclododecane, cyclen) appended with one or two aryl-methyl group(s) (quinolyl-methyl, naphthyl-methyl, and acridinyl-methyl) selectively bind to thymines in a TATA box of the SV40 early promoter region and thus inhibit the binding of a transcriptional factor, TATA binding protein. These Zn2+-cyclen derivatives also act as inhibitors of DNA-targeted enzymes, type I and type II topoisomerases. They also exhibited strong antimicrobial activities for the gram positive bacterial strain. These biochemical and biological properties were compared with those of conventionally established AT-recognizing drugs, distamycin A and DAPI. The Zn2+-cyclen complexes are a new type of small molecular, genetic transcriptional regulation factor. PMID- 10830876 TI - Heavy metal mutagenicity: insights from bioinorganic model chemistry. AB - The mutagenicity of metal species may be the result of a direct interaction with the target molecule DNA. Possible scenarios leading to nucleobase mispairing are discussed, and selected examples are presented. They include changes in nucleobase selectivity as a consequence of alterations in acid-base properties of nucleobase atoms and groups involved in complementary H bond formation, guanine deprotonation, and stabilization of rare nucleobase tautomers by metal ions. Oxidative nucleobase damage brought about by metal species will not be considered. PMID- 10830877 TI - Effect of low temperature on soybean peroxidase: spectroscopic characterization of the quantum-mechanically admixed spin state. AB - A spectroscopic study of soybean peroxidase (SBP) has been carried out using electronic absorption, resonance Raman (RR) and electron paramagnetic resonance (EPR) spectroscopy in order to determine the effects of temperature on the heme spin state. Upon lowering the temperature a transition from high spin to low spin is induced in SBP resulting from conformational changes in the heme cavity, including a contraction of the heme core, the reorientation of the vinyl group in position 2 of the porphyrin macrocycle, and the binding of the distal His to the Fe atom. Moreover, the combined analysis of the data derived from the different techniques at both room and low temperatures demonstrates that at low temperature the quantum-mechanically admixed spin state (QS) of SBP has RR frequencies different from those observed for the QS species at room temperature. PMID- 10830878 TI - Chemical speciation applied to bio-inorganic chemistry. AB - The processes of converting new chemistry into new healthcare, environment friendly, products are considerably facilitated when chemical speciation data are available. These identify causal links between speciation and biological responses based upon specific chemical species rather than total concentrations. The effective roles of bismuth for ulcer treatment, of Ca, Zn, Sn and F in dentifrices, and of zinc species and common cold symptoms are described. A new readily biodegradable ligand for minimising the environmental impact of washing powders, Al and Alzheimer's disease (and the absence of a causal link), and an audit of Cu and Zn flow from wound dressing through wound fluid to healing tissue, are all discussed in detail. PMID- 10830879 TI - In vivo optical/near-infrared spectroscopy and imaging of metalloproteins. AB - A number of medical applications of near-infrared spectroscopy are growing closer to clinical acceptance, and new techniques involving both spectroscopy and imaging are evolving rapidly. In vivo spectroscopy and, more recently, imaging techniques are largely based upon optical electronic transitions involving the metal centers of hemoglobin (blood), myoglobin (muscle) and cytochrome aa3 (mitochondria). The wide variety of near-IR based applications includes heart and stroke research, monitoring cerebral oxygenation of premature babies, and 'functional activation' (response of brain to mental tasks). All of these applications are founded upon changes in hemoglobin O2 saturation; these changes are monitored by following trends in the near-infrared absorptions of deoxyhemoglobin (760 nm) and oxyhemoglobin (920 nm). The same absorptions provide a basis for imaging regional variations in blood oxygenation. This report presents and discusses examples, both from the literature and from our recent work, of near-infrared spectroscopy and imaging in medical applications. PMID- 10830880 TI - Surface-enhanced resonance Raman spectroscopy of porphyrin and metalloporphyrin species in systems with Ag nanoparticles and their assemblies. AB - The advantages of systems with Ag nanoparticles and their assemblies for surface enhanced resonance Raman scattering (SERRS) spectral investigation, detection and determination of porphyrin species are demonstrated. SERRS spectral detection limits of the testing porphyrin species (including porphyrin aggregates) in these systems are shown to be, on average, 10(2)-10(3) lower than detection limits by resonance Raman scattering (RRS). Systems with Ag nanoparticles modified by anionic organosulfur spacers enable us to obtain SERRS spectra of unperturbed cationic porphyrin species. In the case of thiopheneacetate-modified Ag particles prepared by laser ablation, no negative effect of the spacer on the spectral detection limit of the porphyrin was observed. Systems with isolated Ag nanoparticles allow for obtaining SERRS spectra of porphyrin species upon excitation into the Soret electronic absorption band which leads to at least a 10 fold decrease in the detection limit. PMID- 10830881 TI - Infrared spectroscopic studies of nanoscale iron oxide deposits isolated from human thalassemic tissues. AB - Ferritin and hemosiderin isolated from human thalassemic tissues have been characterized by infrared spectroscopy. Spectral features due to both the organic components and the inorganic iron oxyhydroxide have been identified. In particular, spectral evidence for the presence of the goethite (alpha-FeOOH) form of hemosiderin has been obtained in the < 800 cm(-1) range. Various treatments of the hemosiderin isolates result in only small changes in the infrared spectrum indicating the close association of the organic components with the nanoscale iron particles present. PMID- 10830882 TI - Does the haemosiderin iron core determine its potential for chelation and the development of iron-induced tissue damage? AB - Haemosiderin, the major iron storage protein in tissues of iron-loaded tissues shows heterogeneity with respect to both its iron mineralisation product and associated protein. Such mineralisation products have been characterised by a variety of physical techniques including Mossbauer spectroscopy, electron diffraction and EXAFS, and are closely related to the mineral ferrihydrite. A wide range of iron chelators are being developed for the treatment of abnormal haemoglobinopathies, predominantly beta-thalassaemia, which may show greater chelator efficacy for particular mineralisation products of haemosiderin. Even though the tissue iron loadings achieved in different iron-loading syndromes are similar, e.g. naturally occurring iron loading, genetic haemochromatosis and thalassaemia, it is clear that the iron loading in thalassaemic causes extensive damage. The explanation for this could relate to the distribution of iron within different cell types, predominantly reticuloendothelial, its rate of deposition and the mineralisation product of its haemosiderin iron core, goethite. PMID- 10830883 TI - Redox reactions of neurotransmitters possibly involved in the progression of Parkinson's Disease. AB - In Parkinson's Disease the neuromelanin in the substania nigra is known to contain considerably increased amounts of iron suggesting the presence of free, unprotected iron ions during its formation. Iron(II) is known to interact with peroxide via Fenton's reaction producing OH-radicals or ferryl (Fe(IV)) species. This can readily oxidize the neurotransmitter dopamine to the neurotoxic 6 hydroxydopamine (6-OHDA) which is a strong reducing agent. The produced 6-OHDA is, in turn, able to reduce and possibly release iron, as iron(II), from the iron storage protein ferritin. This cycle of events could well explain the development of Parkinson's Disease due to a continuous production of cell damaging species. The contrasting behaviour of 6-OHDA with some other important catecholamines is discussed. PMID- 10830884 TI - The role of free radical reactions with haemoglobin and thalassaemia. AB - It is well known that all living systems depend on iron to transport (haemoglobin), store (myoglobin) and utilize (cytochromes, cytochrome oxide) oxygen for respiration. Iron is an essential component in the active sites of the enzyme that protects against oxidation, such as the iron superoxide dismutase, in bacteria and plants. In normal human plasma almost all iron loading of transferrin is 20-30% maximum. In this presentation we review and summarize recent developments in our understanding of iron transport and storage in living systems. PMID- 10830885 TI - EPR characterization of mono(thiosemicarbazones) copper(II) complexes. Note II. AB - Copper(II) complexes with thiosemicarbazones have been shown to be more active in cell destruction, in the inhibition of DNA synthesis than the uncomplexed ligand. Several derivatives of thiosemicarbazones and their iron and copper complexes have been studied for their cytotoxicity and inhibiting activity against DNA synthesis. In the present work complexes formed in H2O-DMSO solution between copper(II) and the acetophenone thiosemicarbazone (ATSC) and the o aminobenzaldehyde thiosemicarbazone (o-NH2TSC) have been studied. EPR studies have been performed at different pH values and metal-to-ligand ratios. The spectra have been recorded at both room (298 K) and low temperatures (120 K). A possible relationship between structure and activity is attempted on the basis of the EPR data. PMID- 10830886 TI - Effect of pH on the semiquinone radical Q(A)- in CN-treated photosystem II: study by hyperfine sublevel correlation spectroscopy. AB - The semiquinone radical Q(A)- has been studied by electron spin echo envelope modulation (ESEEM) spectroscopy in Photosystem II membranes treated with CN- at various pH values. Two protein 14N nuclei (N(I) and N(II)) were found to be magnetically coupled with the Q(A)- spin. N(I) is assigned to an amide nitrogen from the protein backbone while N(II) is assigned to the amino nitrogen, N(epsilon), of an imidazole. Above pH 8.5 only the N(I) coupling is present while both N(I) and N(II) couplings are present at lower pH values. These results are interpreted in terms of a model based on the structure of the bacterial reaction center and involving two determining factors. First, the non-heme iron, when present, is ligated to the imidazole that H-bonds to one of the Q(A)- carbonyls. This physical attachment of the imidazole to the iron limits the strength of the H-bond to Q(A)-. Second, a pH-dependent group on the protein controls the strength of the H-bonds to Q(A)-. The pKa of this group is around pH 7.5 in CN(-) treated PSII. PMID- 10830887 TI - Trends in NMR chemical shifts and ligand mobility of TcO(V) and ReO(V) complexes with aminothiols. AB - Detailed 1H and 13C NMR studies have been conducted in a series of oxotechnetium and oxorhenium complexes with aminothiol ligands ([SNS][S], [SNN][S], [SNNS]) designed as potential radiopharmaceuticals. The results of these studies in combination with others in the literature show that the oxometal core creates an anisotropic environment and affects the chemical shifts of the coordinated ligandbackbone in a consistent way. Protons oriented towards the oxygen appear deshielded relative to their geminals oriented away from the oxygen. In addition, the direction of a side chain (towards or away from the oxometal core) on the ligand backbone is shown to have a major effect on chemical shifts. The fluxional mobility of the ligand in complexes of the [SNS][S] type was also studied by NMR and the free energy of activation delta G(C)double dagger for the conformational inversion of the ligand was calculated from the temperature dependence of the carbon chemical shifts. delta G(C)double dagger was found to depend on the orientation of the side chain present on the coordinated nitrogen. The energy barrier for the inversion is larger for the oxorhenium complexes than for the analogous oxotechnetium complexes. PMID- 10830888 TI - NMR studies on metal complexes of DNA oligomers. AB - A short overview of NMR spectroscopic applications for the study of metal ion complexes of DNA oligomers is presented. One typical example is given to illustrate the scope of the methods: the NMR structure of a trans-DDP interstrand cross-linked duplex, d(CTCCTG*TGTCTC) x d(GAGATA*AGGAG). The solution structure of this double-stranded DNA oligonucleotide, containing a trans diammineplatinum(II) interstrand cross-link, was determined using two-dimensional nuclear magnetic resonance (2D NMR) and NOE-restrained molecular dynamics and energy minimization refinement. The duplex is a non-palindromic 12/11-mer with a missing central residue in the lower strand and in addition it contains a GT mismatched base pair. The analysis indicated that an interstrand cross-link is established between G6-N7 of the upper strand and A18-N1 of the lower strand. PMID- 10830889 TI - Structural and in vivo studies of metal chelates of Ga(III) relevant to biomedical imaging. AB - The solution chemistry and structure of the complex of the triazamacrocyclic ligand NOTP (1,4,7-triazacyclononane-1,4,7-tris(methylenephosphonate)) with Ga3+ in D2O have been investigated by 1H, 71Ga and 31P NMR spectroscopy. These NMR results show the presence of a 1:1 Ga(NOTP)3- complex, with a highly symmetrical, pseudo-octahedral geometry, possibly with a C3 axis. The 1H spectrum shows that the triazamacrocyclic chelate ring is very rigid, with all the ring protons non equivalent. The complex is stable in aqueous solution in a wide pH range. Its high thermodynamic stability agrees well with previous results from biodistribution and gamma imaging studies in Wistar rats with 67Ga3+ chelates of triaza macrocyclic ligands, which showed that the neutral chelates 67Ga(NOTA) (where NOTA is 1,4,7-triazacyclononane-1,4,7-triacetate) and 67Ga(NOTPME) (where NOTPME is 1,4,7-triazacyclononane-1,4,7-tris(methylenephosphonate monoethylester)) have similar in vivo behaviour, with high stability and rapid renal excretion, but the high negatively charged 67Ga(NOTP)3- has a considerably slower kidney uptake and elimination. PMID- 10830890 TI - NMR studies of metal ion binding to the Zn-finger-like HNH motif of colicin E9. AB - The 134 amino acid DNase domain of colicin E9 contains a zinc-finger-like HNH motif that binds divalent transition metal ions. We have used 1D 1H and 2D 1H-15N NMR methods to characterise the binding of Co2+, Ni2+ and Zn2+ to this protein. Data for the Co2+-substituted and Ni2+-substituted proteins show that the metal ion is coordinated by three histidine residues; and the NMR characteristics of the Ni2+-substituted protein show that two of the histidines are coordinated through their N(epsilon2) atoms and one via its N(delta1). Furthermore, the NMR spectrum of the Ni2+-substituted protein is perturbed by the presence of phosphate, consistent with an X-ray structure showing that phosphate is coordinated to bound Ni2+, and by a change in pH, consistent with an ionisable group at the metal centre with a pKa of 7.9. Binding of an inhibitor protein to the DNase does not perturb the resonances of the metal site, suggesting there is no substantial conformation change of the DNase HNH motif on inhibitor binding. 1H-15N NMR data for the Zn2+-substituted DNase show that this protein, like the metal-free DNase, exists as two conformers with different 1H-15N correlation NMR spectra, and that the binding of Zn2+ does not significantly perturb the spectra, and hence structures, of these conformers beyond the HNH motif region. PMID- 10830891 TI - Multinuclear (13C, 17O, 57Fe) NMR studies of carbonmonoxy heme proteins and synthetic model compounds. AB - 13C, 17O and 57Fe NMR spectra of several carbonmonoxy hemoprotein models with varying polar and steric effects of the distal organic superstructure, constraints of the proximal side, and porphyrin ruffling are reported. Both heme models and heme proteins obey a similar excellent linear delta(13C) versus nu(C O) relationship which is primarily due to modulation of pi-back-bonding from the Fe d(pi) to CO pi* orbital by the distal pocket polar interactions. The lack of correlation between delta(13C) and delta(17O) suggests that the two probes do not reflect a similar type of electronic and structural perturbation. delta(17O) is not primarily influenced by the local distal field interactions and does not correlate with any single structural property of the Fe-C-O unit; however, atropisomerism and deformation of the porphyrin geometry appear to play a significant role. 57Fe shieldings vary by nearly 900 ppm among various hemes and an excellent correlation was found between delta(57Fe) and the absolute crystallographic average displacement of the meso carbon atoms, /Cm/, relative to the porphyrin core mean plane. The excellent correlation between iron-57 shieldings and the average shieldings of the meso carbons of the porphyrin skeleton of TPP derivatives suggests that the two probes reflect a similar type of electronic and structural perturbation which is primarily porphyrin ruffling. PMID- 10830892 TI - Bacterial cytochrome c nitrite reductase: new structural and functional aspects. AB - Cytochrome c nitrite reductase catalyzes the six-electron reduction of nitrite to ammonia as a key step within the biological nitrogen cycle. Most recently, the crystal structure of the soluble enzyme from Sulfurospirillum deleyianum could be solved to 1.9 A resolution. This set the basis for new experiments on structural and functional aspects of the pentaheme protein which carries a Ca(2+) ion close to the active site heme. In the crystal, the protein was a homodimer with ten hemes in very close packing. The strong interaction between the nitrite reductase monomers also occurred in solution according to the dependence of the activity on the protein concentration. Addition of Ca(2+) to the enzyme as isolated had a stimulating effect on the activity. Ca(2+) could be removed from the enzyme by treatment with chelating agents such as EGTA or EDTA which led to a decrease in activity. In addition to nitrite, the enzyme converted NO, hydroxylamine and O methyl hydroxylamine to ammonia at considerable rates. With N2O the activity was much lower; most likely dinitrogen was the product in this case. Cytochrome c nitrite reductase exhibited a remarkably high sulfite reductase activity, with hydrogen sulfide as the product. A paramagnetic Fe(II)-NO, S = 1/2 adduct was identified by rapid freeze EPR spectroscopy under turnover conditions with nitrite. This potential reaction intermediate of the reduction of nitrite to ammonia was also observed with PAPA NONOate and Spermine NONOate. PMID- 10830893 TI - A search for beta-lactamase in chlamydiae, mycoplasmas, planctomycetes, and cyanelles: bacteria and bacterial descendants at different phylogenetic positions and stages of cell wall development. AB - Bacteria from different phylogenetic positions such as chlamydiae, mycoplasmas, planctomycetes and also endosymbiotic murein-containing cyanelles were investigated for the production of beta-lactamases. No beta-lactamase activity was found in bacteria lacking murein such as Chlamydia pneumoniae, Mycoplasma pneumoniae, Pirellula marina and Planctomyces maris. In the murein-containing cyanelles of Cyanophora paradoxa no beta-lactamase activity could be detected. PMID- 10830894 TI - Endochitinase gene-based phylogenetic analysis of Trichoderma. AB - DNA sequences of the single-copy gene coding for the 42 kDa endochitinase enzyme (EC 3.2.1.14) were used for phylogenetic analysis in Trichoderma. A set of 12 primers was developed and the entire gene was sequenced for 16 strains, and nucleotide and deduced amino acid sequences were compared to data from GenBank for additional Trichoderma strains. Analysis of the sequences revealed parsimony informative variation from 2.4 to 43.6% depending on the part of the gene (exons/introns) and the taxonomic level considered. Results are discussed in comparison to previous data from ITS-1 and ITS-2 rDNA sequencing and suggest the 42 kDa endochitinase gene as a potential molecular marker for reconstructing phylogenetic relationships in the genus Trichoderma at species level. PMID- 10830895 TI - Bacillus thuringiensis soil populations naturally occurring in the Ryukyus, a subtropic region of Japan. AB - Of 809 soil samples collected from the seven islands of the Ryukyus, Japan, 107 samples (13.2%) contained Bacillus thuringiensis. The frequency of B. thuringiensis among the B. cereus group was 1.1% (235/21842) on the average. The B. thuringiensis soil populations of the Ryukyus consisted of more than 22 H serogroups. The predominant H serotype was the H5ac/21 (serovar canadensis/colmeri), followed by the H3ad (serovar sumiyoshiensis) and H16 (serovar indiana). Geographically, most widely distributed H serogroups were the H16 and H10ac (serovar londrina); the former was recovered from five islands and the latter from three islands. Parasporal inclusions of the isolates were morphologically heterogeneous, roughly grouped into four categories: bipyramidal/cuboidal, spherical/ovoid, irregularly-pointed, and irregular-shaped. About 53% of the isolates formed spherical to ovoid parasporal inclusions. None of the isolates exhibited larvicidal activity against the silkworm, Bombyx mori. Only four isolates belonging to four different serotypes killed larvae of the mosquito, Aedes aegypti. These mosquito-specific isolates all produced spherical parasporal inclusions. PMID- 10830896 TI - Larval susceptibility of the diamondback moth, Plutella xylostella (Lepidoptera: Plutellidae), to Bacillus thuringiensis H serovars isolated in Japan. AB - A total of 1700 Japanese strains of Bacillus thuringiensis, belonging to at least 47 H serogroups, were examined for insecticidal activity against larvae of the diamondback moth, Plutella xylostella. The high-level toxicity was associated with 612 isolates (36.0%). Of these, 608 isolates (99.3%) fell into 13 H serogroups belonging to the low-numbered H serotypes, H1-H10. Conversely, most isolates belonging to the high-numbered serotypes (>H10) had little or no larvicidal activity; only one isolate of the serovar japonensis H23 was active. P xylostella larvae were susceptible to 89.8% of the serovar morrisoni H8a:8b strains and 85.7% of galleriae H5a:5b strains. High values of 60-80% were also obtained in six serovars (thuringiensis H1, alesti H3a:3c, kurstaki H3a:3b:3c, kenyae H4a:4c, aizawai H7, and tolworhi H9), while relatively low values of <60% in two other common serovars, sotto H4a:4b and darmstadiensis H10a:10b. Five selected isolates, belonging to H serovars other than kurstaki and aizawai, were 10-60 times less toxic than the reference strain HD-1 (serovar kurstaki). Parasporal inclusion proteins of these strains were immunologically unrelated to those of the strain HD-1 and the aizawai type strain. PMID- 10830897 TI - Effect of magnesium ions on fermentative and respirative functions in Pichia stipitis under oxygen-restricted growth. AB - Mg2+ level affected growth, xylitol and ethanol production by P. stipitis grown under microaerophilic conditions. Low Mg2+ level (1 mM) directed the C flux from ethanol to xylitol, with no effect on xylose consumption rate. The addition of pyrazole, an alcohol dehydrogenase (ADH) inhibitor, had the same effect, even in conditions of Mg2+ excess (4 mM), indicating a negative interaction between ADH and Mg2+ ions (p << 0.01). Cells grown either with pyrazole or Mg limitation increased their intracellular NADH concentration about 3 times, but displayed no significant differences in ADH specific activities (1,000 U/mg protein, +/- 10%). In contrast, no interaction was measured between Mg and antimycin A, excluding the possibility that Mg2+ limitation interferes with respiration. PMID- 10830898 TI - Production of citric acid from starch-hydrolysate by Aspergillus niger. AB - The present investigation explored the possible use of a rarely used agro industrial by-product, maize starch-hydrolysate, for economic production of citric acid. To achieve this, seventeen strains of Aspergillus niger were screened for their capacity to produce citric acid using starch-hydrolysate as a substrate. The most efficient strain, ITCC-605 was selected for further improvement in citric acid content by mutation. Mutants developed by treatment with EMS and UV, singly and in combination, produced citric acid in the range of 0.51-64.7 g kg(-1) of glucose consumed. The mutant UE-1 produced the maximum citric acid which was about 130 times more than that produced by the parent strain, ITCC-605. For further increase in citric acid production from this substrate, the cultural conditions were optimized: concentration of starch hydrolysate, 15% (glucose equivalent); ammonium nitrate, 0.25%; KH2PO4, 0.15%; nicotinic acid, 0.0001% and initial pH of 2.0. Under these conditions, the mutant strain UE-1 yielded 490 g citric acid kg(-1) of glucose consumed in 8 days of incubation at 30 degrees C. The productivity of 341 mgl(-1)h(-1) corresponded to 49% substrate conversion to citric acid. PMID- 10830899 TI - Transient inhibition of glucose utilization by erythrocytes during the acute stage of typhoid fever. AB - The exact reason for hemolysis of glucose-6-phosphate dehydrogenase-deficient (G6PD) erythrocytes in patients with typhoid fever is unknown. Therefore, glucose utilization by normal and G6PD-deficient erythrocytes was measured during incubation with plasma of healthy controls as well as from patients in acute or recovery stages of typhoid fever. Glucose utilization in normal and G6PD deficient erythrocytes significantly decreased compared to the controls when incubated with plasma of patients with acute typhoid fever, which normalized to the baseline after recovery from typhoid fever, suggesting an acquired alteration in G6PD enzyme properties by Salmonella typhi or its endotoxins. PMID- 10830900 TI - Higher prevalence of Alternaria spp. in marine and river waters than in potable samples. AB - Alternaria species have been recognized as important cause of allergic diseases and are considered opportunistic pathogens for immunocompromised individuals. In order to assess the contribution of waters to the spread of Alternaria, we examined 390 water samples of various origin for the presence of these microorganisms in parallel with the standard pollution indicator bacteria, i.e. total coliforms, faecal coliforms and enterococci. Alternaria spp. were isolated from 42 out of 196 (21.4%) marine water samples, from 13 out of 68 (19.1%) river samples, whereas out of 126 potable waters only two (1.6%) were found positive. The incidence of Alternaria was significantly higher (p < 0.001) in marine and river samples than in potable waters. The mean values of the colony-forming units of the pollution indicator bacteria did not significantly differ between Alternaria-positive and Alternaria-negative samples. PMID- 10830901 TI - Molecular and microscopical detection of aster yellows phytoplasma associated with infected parsnip. AB - Typical phytoplasma yellows symptoms were observed in parsnip (Pastinaca sativa L.) plants grown around Edmonton, Alberta, Canada. Examination of ultrathin sections of leaf midribs by electron microscopy revealed numerous phytoplasma bodies localized in the phloem cells. DNA extracted from the infected leaves was amplified with a 16S rDNA universal primer pair P1/P6 giving the expected PCR product of 1.5 kb. The phytoplasma was confirmed as a member of the aster yellows (AY) group by amplification with the specific primer pair R16(1)/F1/R1 that was designed on the basis of AY phytoplasma 16S rDNA sequences. In the nested PCR assays, the expected DNA fragment of 1.1 kb was amplified with this specific primer set. Similar restriction patterns were found for the 1.1 kb PCR products of the phytoplasma isolated from parsnip and an AY phytoplasma control after digestion with restriction endonucleases AluI, HhaI, KpnI and RsaI. This is the first reported observation of aster yellows in parsnip in Canada. PMID- 10830902 TI - Physico-chemical parameters influencing DNase activity of the cyanobacterium Spirulina platensis. AB - The effects of temperature, Mg2+, EDTA concentration and rinsing on extra- and intra-cellular DNase activity of Spirulina platensis strain SSP-14, were investigated. The results indicate that the tested strain contains very high extra- and intracellular DNase activity, which actually hinders the transfer of foreign gene(s) to S. platensis, a cyanobacterium with multiple economic potentials. The extracellular DNase activity could easily be removed by rinsing the cells with Zarrouk medium more than once. The intracellular DNase activity could also be inhibited by (1) removal of Mg2+, (2) maintaining EDTA concentration above 1 mmol l(-1), and (3) manipulating below 0-4 degrees C, during all the incubation procedures. We suggest that, by using one or more of, or combining, all those experimental conditions, the chances of foreign DNA attempted to be introduced into S. platensis without being digested would be increased. PMID- 10830903 TI - The role of ferrioxamine E in pre-enrichment medium for determining Salmonella in environmental samples according to ISO method. AB - The effect of a siderophoric compound, ferrioxamine E, in the pre-enrichment broth on determining of Salmonella infantis in environmental samples was tested with combination of various pre-enrichment times and enrichment temperatures of 37 and 43 degrees C. Ferrioxamine E slightly improved the determination efficiency of this bacterium but the pre-enrichment time could not be reduced below 17 hours. The enrichment temperature of 43 degrees C was better than of 37 degrees C. The mixing ratios of 1:100 or 1:1000 for samples and pre-enrichment broth were more successful than the ratio of 1:10 as recommended by ISO. PMID- 10830904 TI - Characterization of the human complex I NDUFB7 and 17.2-kDa cDNAs and mutational analysis of 19 genes of the HP fraction in complex I-deficient-patients. AB - Deficiency of NADH:ubiquinone oxidoreductase, the first enzyme complex of the mitochondrial respiratory chain, is one of the most frequent causes of human mitochondrial encephalomyopathies. A relatively small percentage of human complex I deficiency is associated with mitochondrial DNA mutations. cDNA characterization and mutational analysis of the structural complex I genes in 19 complex I-deficient patients, in whom common mtDNA mutations have been excluded, has so far revealed five patients with alterations in evolutionary conserved nuclear-encoded proteins. In order to complete our knowledge about the expected 36 structural nuclear complex I genes, we characterized the NDUFB7 and the 17.2 kDa cDNA sequences of the hydrophobic (HP) fraction of the complex. Subsequently, we screened all subunits of this fraction for the presence of mutations in those 14 patients of our initial patient cohort in whom the underlying genetic cause had not been elucidated. Strikingly, no pathogenic mutations were found in the HP subunits that would explain the complex I deficiency in our patients. Other strategies are needed to unravel proteins involved in the pathogenesis of the complicated cellular network of transcription until correct assemblage of complex I. PMID- 10830905 TI - Individuals with abnormal phenotype and normal G-banding karyotype: improvement and limitations in the diagnosis by the use of 24-colour FISH. AB - The simultaneous identification, by fluorescence in situ hybridisation (FISH), of each chromosome in a distinct colour became feasible a few years ago. The key question in the application of this and many other developments in molecular cytogenetics to clinical situations is whether the results add significant further information that is relevant to the diagnosis. So far, limited data exist regarding how much improvement the technique brings to the diagnosis of phenotypically abnormal individuals in whom no abnormalities have been detected by conventional G-banding analysis. Because of the lack of a conclusive diagnosis, genetic counselling, estimation of recurrence risk and prenatal diagnosis of these individuals and their relatives is problematic. We report a study with 24-colour whole-chromosome painting of 10 familial and 11 isolated cases with abnormal phenotypes and normal G-banding karyotypes. Previously undetected unbalanced translocations were revealed in two cases. The value and current cost-effectiveness of multicolour FISH for cytogenetic diagnosis is discussed. PMID- 10830906 TI - High frequency hearing loss correlated with mutations in the GJB2 gene. AB - Genetic hearing impairment affects approximately 1/2000 live births. Mutations in one gene, GJB2, coding for connexin 26 cause 10%-20% of all genetic sensorineural hearing loss. Mutation analysis in the GJB2 gene and audiology were performed on 106 families presenting with at least one child with congenital hearing loss. The families were recruited from a hospital-based multidisciplinary clinic, which functions to investigate the aetiology of sensorineural hearing loss in children and which serves an ethnically diverse population. In 74 families (80 children), the aetiology was consistent with non-syndromic recessive hearing loss. Six different connexin 26 mutations, including one novel mutation, were identified. We show that GJB2 mutations cause a range of phenotypes from mild to profound hearing impairment and that loss of hearing in the high frequency range (4000 8000 Hz) is a characteristic feature in children with molecularly diagnosed connexin 26 hearing impairment. We also demonstrate that this type of audiology and high frequency hearing loss is found in a similar-sized group of deaf children in whom a mutation could only be found in one of the connexin 26 alleles, suggesting connexin 26 involvement in the aetiology of hearing loss in these cases. In our study of the M34T mutation, only compound heterozygotes exhibited hearing loss, suggesting autosomal recessive inheritance. PMID- 10830907 TI - Structure and mutation analysis of the gene encoding DNA fragmentation factor 40 (caspase-activated nuclease), a candidate neuroblastoma tumour suppressor gene. AB - We have characterised the DFFB gene, encoding the active subunit of the apoptotic nuclease DNA fragmentation factor (DFF40). DFFB maps to 1p36, near the imprinted putative tumour suppressor gene TP73. The DFFA gene (encoding the inhibitory DFF45 subunit) also maps to 1p36.2-36.3, and we show by FISH that DFFB lies distal to DFFA. We have also mapped a processed DFFB pseudogene to chromosome 9. DFFB itself has seven coding exons spanning 10 kb. Exhaustive mutation screening of 41 neuroblastomas and other tumours in which a 1p36 tumour suppressor gene is implicated showed no tumour-specific mutations. A coding region polymorphism was used to demonstrate uniformly biallelic expression in human fetal DFFB transcripts. Since the putative neuroblastoma tumour suppressor gene in distal 1p36 is predicted to be maternally expressed, the lack of imprinting and absence of somatic mutations in DFFB indicate that it is probably not the neuroblastoma tumour suppressor gene. PMID- 10830908 TI - CYP21 mutations in Brazilian patients with 21-hydroxylase deficiency. AB - Congenital adrenal hyperplasia caused by 21-hydroxylase deficiency is a common autosomal recessive disorder resulting from mutations in the 21-hydroxylase (CYP21) gene. To develop a strategy to screen for the most commonly occurring CYP21 mutations in Brazil, we performed molecular genotype analysis on 73 children with CAH representing 71 unrelated families. The techniques used for CYP21 molecular genotype analysis were: restriction fragment length polymorphism, single-strand conformational polymorphism, allele-specific oligonucleotide hybridization, allele-specific polymerase chain reaction amplification, and heteroduplex analyses. Mutations were identified on all but eight affected alleles. The intron 2 splicing mutation was the most frequently identified mutation. Screening for the most common mutations detected at least one mutation on 132/142 (93%) alleles. Multiple CYP21 mutations were detected on 16.2% of alleles. The high frequency of multiple mutations on a single allele emphasizes the importance of thorough and accurate molecular genotype analysis of the complex CYP21 locus. PMID- 10830909 TI - Lipoprotein lipase gene variation is associated with adipose tissue lipoprotein lipase activity, and lipoprotein lipid and glucose concentrations in overweight postmenopausal women. AB - Adipose tissue lipoprotein lipase (LPL) activity is under strong genetic control in both mice and humans. This study determines whether common DNA variation in the LPL gene (PvuII and HindIII polymorphisms) is associated with adipose tissue LPL activity and metabolic risk factors in a homogeneous population of 75 overweight postmenopausal women (body mass index >25 kg/m2; age: 51-69 years old). The allele frequencies for the presence of the cut-sites for LPL HindIII and PvuII were 0.71 and 0.49, respectively. There were no associations between the HindIII polymorphism and any of the measured variables. Age, body mass index, percent body fat, waist-hip ratio, visceral and subcutaneous fat area, and gluteal (GLT) and abdominal (ABD) adipocyte size did not differ by LPL PvuII genotype. However, adipose tissue LPL activity at both GLT and ABD sites was higher in women without the LPL PvuII cut-site (-/-) compared with women who were heterozygous (+/-) or homozygous (+/+) for the cut-site (P<0.05). Total and LDL cholesterol were lower in women without the LPL PvuII cut-site (-/-) compared with women who were heterozygous or homozygous for the cut-site (P<0.05), whereas triglyceride and HDL levels were similar between LPL PvuII genotypes. Fasting glucose, but not insulin, was lower in women without the LPL PvuII cut-site (-/ ). These data suggest that the LPL PvuII polymorphism is a possible marker for a functional mutation that is found in the LPL gene and that alters LPL activity in older overweight women. PMID- 10830910 TI - Genotype-phenotype correlations in families with deletions in the von Hippel Lindau (VHL) gene. AB - Von Hippel-Lindau (VHL) disease is a hereditary tumor syndrome characterized by predisposition for bilateral and multi-centric hemangioblastoma in the retina and central nervous system, pheochromocytoma, renal cell carcinoma, and cysts in the kidney, pancreas, and epididymis. We describe five families for which direct sequencing of the coding region of the VHL gene had failed to identify the family specific mutation. Further molecular analysis revealed deletions involving the VHL gene in each of these families. In four families, partial deletions of one or more exons were detected by Southern blot analysis. In the fifth family, FISH analysis demonstrated the deletion of the entire VHL gene. Our results show that (quantitative) Southern blot analysis is a sensitive method for detecting germline deletions of the VHL gene and should be implemented in routine DNA diagnosis for VHL disease. Our data support the previously established observation that families with a germline deletion have a low risk for pheochromocytoma. Further unraveling of genotype-phenotype correlations in VHL disease has revealed that families with a full or partial deletion of the VHL gene exhibit a phenotype with a preponderance of central nervous system hemangioblastoma. PMID- 10830911 TI - Fine mapping of the neurally expressed gene SOX14 to human 3q23, relative to three congenital diseases. AB - Members of the Sox gene family encode transcription factors that have diverse and important functions during development. We have recently described the cloning of chick and mouse Sox14 and the expression of these genes in a population of ventral interneurons in the embryonic spinal cord. We report here the cloning and sequencing of the human orthologue of Sox14. Human SOX14 shows remarkable sequence conservation compared with orthologues from other vertebrate species and probably mirrors the expression of these genes in the developing brain and spinal cord. Using radiation hybrid mapping and fluorescence in situ hybridisation, we have localised SOX14 close to the sequence tagged site D3S1576 on human chromosome 3q23. Three congenital disorders have been localised to this region: blepharophimosis-ptosis-epicanthus inversus syndrome (BPES), Charcot-Marie-Tooth neuropathy type IIB (CMT2B) and Mobius syndrome type 2 (MBS2). We have found that SOX14 is unlikely to be involved in any of these disorders because of the position of SOX14 proximal to a BPES breakpoint and the lack of SOX14 coding region alterations in BPES, CMT2B and MBS2 patients. PMID- 10830912 TI - Association studies of 33 single nucleotide polymorphisms (SNPs) in 29 candidate genes for bronchial asthma: positive association a T924C polymorphism in the thromboxane A2 receptor gene. AB - Although intensive studies have attempted to elucidate the genetic background of bronchial asthma (BA), one of the most common of the chronic inflammatory diseases in human populations, genetic factors associated with its pathogenesis are still not well understood. We surveyed 29 possible candidate genes for this disease for single nucleotide polymorphisms (SNPs), the most frequent type of genetic variation, in genomic DNAs from Japanese BA patients. We identified 33 SNPs, only four of which had been reported previously, among 14 of those genes. We also performed association studies using 585 BA patients and 343 normal controls for these SNPs. Of the 33 SNPs tested, 32 revealed no positive association with BA, but a T924C polymorphism in the thromboxane A2 receptor gene showed significant association (chi2=4.71, P=0.030), especially with respect to adult patients (chi2=6.20, P=0.013). Our results suggest that variants of the TBXA2R gene or some nearby gene(s) may play an important role in the pathogenesis of adult BA. PMID- 10830913 TI - Association of butyrylcholinesterase K variant with cholinesterase-positive neuritic plaques in the temporal cortex in late-onset Alzheimer's disease. AB - In confirmed late-onset (>65 years) Alzheimer's disease, we found a greater load, both of overall neuritic plaques and of cholinesterase-positive neuritic plaques, in the temporal cortex of carriers of the butyrylcholinesterase K variant (BCHE K) aged <80 years than of all other patients. The differences were most striking in the case of cholinesterase-positive neuritic plaques. Among BCHE-K carriers, densities of such plaques were over six times higher in patients <80 years at death than in those >80 years (P=0.01). Furthermore, in subjects <80 years, BCHE K carriers had nearly six-fold greater densities of these plaques than non carriers (P=0.009). We consider three potential explanations for these findings: that the K variant binds more readily to plaque constituents, that it promotes fibril formation or that it induces aberrant neurite growth. PMID- 10830914 TI - Multiple origins of Tibetan Y chromosomes. AB - The genetic origin of Tibetans was investigated using Y chromosome markers. A total of three populations were studied, two from central Tibet speaking central Tibetan and one from Yunnan speaking Kham. Two dominant paternal lineages (>80%) were identified in all three populations with one possibly from central Asia (YAP+) and the other from east Asia (M122C). We conclude that Tibetan Y chromosomes may have been derived from two different gene pools, given the virtual absence of M122C in central Asia and YAP+ in east Asia, with drift an unlikely mechanism accounting for these observations. PMID- 10830915 TI - Age of the intronic GAA triplet repeat expansion mutation in Friedreich ataxia. AB - Friedreich ataxia (FRDA), the most frequently inherited ataxia, is due in the vast majority of cases to a large expansion of an intronic GAA repeat. Using linkage disequilibrium analysis based on haplotype data of seven polymorphic markers close to the frataxin gene, the age of FRDA founding mutational event(s) is estimated to be at least 682+/-203 generations (95% confidence interval: 564 801 g), a dating which is consistent with little or no negative selection and provides further evidence for an ancient spread of a pre-mutation (at-risk alleles) in western Europe. PMID- 10830916 TI - Sarcoplasmic reticulum Ca2+ regulatory protein gene expression in human right atrium under hemodynamic overload. AB - Sarcoplasmic reticulum (SR) Ca2+-adenosine triphosphatase (ATPase) mRNA expression is reduced in the failing human myocardium. However, it is not known whether SR Ca2+-regulatory protein gene expression is altered in human myocardial tissue subjected to pressure overload or volume overload. We sought to determine whether SR Ca2+-regulatory protein gene expression is altered in human atrial tissue subjected to mechanical overload. We obtained right atrial myocardial tissue (about 250mg) at open-heart surgery from three groups of patients: no hemodynamic overload to the right atrium (control group; 12 patients), atrial septal defect (ASD group; 8 patients), and tricuspid regurgitation (TR group; 7 patients). We measured the myocyte size, the area of interstitial fibrosis, SR Ca2+,-ATPase, and ryanodine receptor mRNA abundance. The isolated cardiocyte area and the percent area of interstitial fibrosis were in the order TR > ASD > control (P < 0.05). The SR Ca2+-ATPase mRNA level in TR was significantly decreased (P = 0.004) compared with the control, whereas in the ASD group it did not differ significantly from control. There were no significant differences in ryanodine receptor mRNA levels among the three groups. SR Ca2+-ATPase gene expression was downregulated in human atrial tissue with TR but not in ASD, which might have resulted from the differences in the degree and/or the type of hemodynamic overload to the myocardium. PMID- 10830917 TI - Na+-K+ pump inhibition caused by chronic amiodarone in guinea pig myocardium. AB - Although it is known that amiodarone inhibits myocardial Na+-K+ pump activity, the potency and the time course of this inhibition are unknown. The aim of this study was to investigate these aspects with reference to digoxin, using guinea pigs treated with either intraperitoneal amiodarone (20mg/kg per week, up to 12 weeks, n = 26) or the same amount of vehicle as a control (n = 24). ECG recording and microelectrode experiments were conducted every 2 weeks. QT interval corrected by heart rate and action potential duration were prolonged as a function of the time of exposure to amiodarone. Hyperpolarization observed immediately after the overdrive (1.0Hz) termination or K+-replenishment following K+-depletion in the presence of 0.1mM Ba2+ was compared in the amiodarone-treated and untreated groups, as an index of the Na+-K+ pump activity. The resting membrane potential recovery from overdrive-induced depolarization was slower and the amplitude of K+-induced hyperpolarization was smaller in the amiodarone treated group than in the untreated group. These changes were evident as the chronic amiodarone treatment progressed, although the changes in these parameters were greater in the case of acute application of 50 microM digoxin. In conclusion, this study indicates that treatment with amiodarone for longer than several weeks moderately inhibits the myocardial Na+-K+ pump. PMID- 10830918 TI - Effect of olprinone, a phosphodiesterase III inhibitor, on arterial wall distensibility: differentiation between aorta and common carotid artery. AB - Although the effects of phosphodiesterase III (PDE III) inhibitors as vasorelaxants have been well documented, there are only few data on the wall response of different arteries. We evaluated the artery-specific effect of olprinone (OP), one of the PDE III inhibitors, on the major branches of human arteries and peripheral circulation. In 14 healthy subjects (average age: 57.5 +/ 21.2 years), systolic and diastolic diameters (Ds and Dd, respectively) and the time velocity integral (VI) of flow velocity patterns were measured by M-mode and Doppler echocardiography in the carotid artery (CA), the ascending aorta (asAo), the abdominal aorta (abAo), and the left ventricular outflow tract. Blood pressure (BP) was simultaneously measured using a cuff sphygmomanometer. Measurements were taken before and 20min after a bolus injection of OP (0.2 microg/kg). Distensibility (Ds - Dd), stiffness parameter beta (In(systolic BP/diastolic BP)/(Ds/Dd - 1)), cardiac output (CO: (Flow Area) x VI x HR at left ventricular outflow), selective flow volume (FV: (Flow Area) x VI x HR at CA or abAo), and vascular resistance (VR: mean BP/(CO or FV)) were then calculated. The distensibility increased significantly after OP administration (P = 0.0015), but that of the asAo or abAo did not change. Although there was a significant increase in CO (P = 0.001) and a significant decrease in systemic VR (P = 0.001) following OP administration, the FV and VR of both CA and abAo did not change significantly. The selectiveness of the effect of OP was demonstrated in terms of the CA wall distensibility. This was thought to be attributable to the differences in the structural components or the reactivity of smooth muscle cells to OP. PMID- 10830919 TI - Validity of the right ventricular Doppler index for assessment of severity of congestive heart failure in patients with dilated cardiomyopathy. AB - This study was designed to clarify the validity of Doppler indices for assessment of the severity of congestive heart failure in patients with dilated cardiomyopathy. Left ventricular (LV) and right ventricular (RV) Doppler indices, defined as the sum of isovolumic contraction time and relaxation time divided by the ejection time, were obtained in 30 normal subjects and 35 patients with ischemic or idiopathic dilated cardiomyopathy. We analyzed the correlation between New York Heart Association (NYHA) functional class, right-sided cardiac pressure, and the Doppler indices. Patients with dilated cardiomyopathy revealed significantly higher LV and RV Doppler indices than normal subjects; however, the LV Doppler index did not correlate significantly with NYHA functional class or right-sided cardiac pressure. On the other hand, the RV Doppler index correlated significantly with NYHA functional class (rs = 0.83, P < 0.001), pulmonary capillary wedge pressure (r = 0.80, P < 0.001), pulmonary artery systolic pressure (r = 0.81, P < 0.001) and pulmonary artery diastolic pressure (r = 0.77, P < 0.001). Both RV isovolumic contraction and relaxation time also correlated significantly with right-sided cardiac pressure. In 12 patients who improved from NYHA III-IV to I-II after treatment, the RV Doppler index decreased from 0.84 +/- 0.25 to 0.33 +/- 0.10 (P < 0.001), while the LV Doppler index did not change significantly (0.72 +/- 0.27 to 0.68 +/- 0.23, P = 0.53). These results suggest that the RV Doppler index is useful for assessment of the severity of congestive heart failure in patients with dilated cardiomyopathy. PMID- 10830920 TI - Correlation of anatomic and hemodynamic features with aortic valve leaflet deformity in doubly committed subarterial ventricular septal defect. AB - The records of 153 patients with doubly committed subarterial ventricular septal defect (DCVSD) who underwent intracardiac repair were analyzed to evaluate factors responsible for aortic valve leaflet deformity. The patients were divided into two groups according to their echocardiographic and angiographic features as well as anatomic findings at operation: DCVSD without (17/153, 11.1%) and with arterial valve offsetting (136/153, 88.9%). Aortic regurgitation (AR) was much more prevalent in the patients with (50.0%) than in those without leaflet deformity (2.2%, P < 0.01). Arterial valve offsetting is one of the major contributing factors to the development of leaflet deformity, accounting for 5.9% in the patients without offsetting and 46.3% in those with offsetting (P < 0.01). Among the patients with arterial valve offsetting, the pulmonary-to-systemic pressure ratio was significantly higher (P < 0.01) in the patients without (0.76 +/- 0.14) than in those with leaflet deformity (0.36 +/- 0.12), suggesting that pulmonary hypertension might prevent the aortic valve leaflet from prolapsing in DCVSD. In addition, increased severity of aortic valve leaflet deformity and subsequent AR were observed with increasing age. These results suggest that aging and the presence of arterial valve offsetting as well as the absence of pulmonary hypertension might be factors responsible for aortic valve leaflet deformity and subsequent AR in DCVSD. The anatomic and hemodynamic features in DCVSD have a great impact on the development of aortic valve leaflet deformity and subsequent AR. PMID- 10830921 TI - Characterization of von Willebrand factor in primary pulmonary hypertension. AB - The aim of this study was to determine the value of von Willebrand factor (vWF), a well-characterized endothelial cell protein secretion, as a marker for prognosis in patients with primary pulmonary hypertension (PPH). Venous and arterial blood samples were obtained from 18 clinically diagnosed PPH patients and 12 case controls matched for age and sex. Plasma vWF antigen was determined by enzyme-linked immunosorbent assay (ELISA). The patients' multimeric vWF pattern was analyzed by sodium dodecylsulfate (SDS)-agarose-acrylamide electrophoresis, Western blot, and densitometric analysis. vWF sialic acid content was determined by a lectin-based ELISA. The PPH patients showed a higher content of vWF antigen in venous (P = 0.0026) and arterial (P = 0.0094) blood samples than controls. The mean vWF sialic acid content of the PPH patients corresponded to 37.7% of the mean value for the control group. On the basis of the hemodynamic response to vasodilator trial, the PPH patients were grouped as responders or nonresponders. The latter group showed a significantly higher plasma vWF antigen antecubital vein/radial artery ratio, an increased number of unusually large vWF multimers, and a diminished content of vWF sialic acid in comparison with the first group. We believe that our results establish the nature of vWF alterations that are related to endothelial cell damage in patients with primary pulmonary hypertension and that this could be of value when establishing the prognosis in this group of patients. PMID- 10830924 TI - Europe agrees to boost Internet networks used by researchers PMID- 10830923 TI - Benefits of increased public participation. PMID- 10830922 TI - Vessel size-dependent expression of intermediate-sized filaments, calponin, and h caldesmon in smooth muscle cells of human coronary arteries. AB - The arterial media is composed of a heterogeneous population of smooth muscle cells (SMCs). Recently, the properties of SMCs were observed to be heterogeneous not only among individual cells but also among arteries of the same vascular bed. To test the hypothesis that a site-specific heterogeneity exists in the SMCs of human coronary arteries, we examined the expression of desmin, vimentin, calponin, and high-molecular-weight (h-) caldesmon in arteries of various sizes. Specimens of arteries were obtained at autopsy from 12 patients: 6 adults (67 +/- 4 years old); 3 younger adults (26 +/- 2 years old); and 3 neonates. The size of the arteries was estimated by the number of SMC layers of the media. The expression was compared in SMCs of large arteries (>10 layers in adults, >5 layers in neonates), medium-sized arteries (5-10 layers in adults, 3-5 SMC layers in neonates), and small arteries (<3 layers). In adults, the percentage of arteries positive for desmin was lower in the small (17% +/- 3%) and medium-sized arteries (44% +/- 12%) than in the large arteries (94% +/- 6%) (P < 0.01). The percentage of arteries positive for calponin was also lower in the small (18% +/- 2%) and medium-sized arteries (66% +/- 5%) than in the large arteries (100%) (P < 0.01). The percentage for vimentin and h-caldesmon did not differ among large, medium-sized, and small arteries. These observations in adults were similar to those in younger adults or neonates. The phenotypes of medial SMCs are vessel size-dependent in human coronary arteries. This finding should be important for understanding the site-specific characteristics of vascular function in the regulation of myocardial perfusion or those of vascular responses to environmental changes. PMID- 10830925 TI - Congo war increases threat to bonobo research. PMID- 10830926 TI - NIH takes charge of chimps infected in experiments. PMID- 10830927 TI - Cheaper AIDS drugs due for Third World.. PMID- 10830928 TI - But South Africa voices reservations. PMID- 10830929 TI - Geneticist from Baylor named as new head of UK's Sanger Centre. PMID- 10830930 TI - Researchers take a gamble on the human genome. PMID- 10830931 TI - Los Alamos labs are safe from fire PMID- 10830932 TI - Gene bank to offer family album of mammals. PMID- 10830933 TI - Japan calls for open access to human genome data. PMID- 10830935 TI - India pushes ahead with plant database. PMID- 10830934 TI - Greens persuade Europe to revoke patent on neem tree... PMID- 10830936 TI - Reservoirs dog AIDS therapy. PMID- 10830937 TI - AIDS dissidents aren't victims--but the people their ideas kill will be. PMID- 10830939 TI - More is less. Economists and governments lag decades behind Derek Price's thinking PMID- 10830938 TI - Bureaucracy strangles Latin American research PMID- 10830940 TI - Consequences. Perhaps the biotech sector should have employed more philosophers PMID- 10830941 TI - Recounting a genetic story. PMID- 10830942 TI - The comet no one saw PMID- 10830943 TI - Cancer. Checkpoint for invasion. PMID- 10830944 TI - Palaeoceanography. Nutrients in the glacial balance PMID- 10830945 TI - Bioluminescence illuminated. PMID- 10830946 TI - Materials science. Plastic sandwiches a la carte PMID- 10830947 TI - Guiding the swing in golf putting. PMID- 10830948 TI - Evolution of lifespan in C. elegans. PMID- 10830949 TI - How cicadas interpret acoustic signals. PMID- 10830950 TI - Transfection of cells by immunoporation. PMID- 10830951 TI - Lateral gene transfer and the nature of bacterial innovation. AB - Unlike eukaryotes, which evolve principally through the modification of existing genetic information, bacteria have obtained a significant proportion of their genetic diversity through the acquisition of sequences from distantly related organisms. Horizontal gene transfer produces extremely dynamic genomes in which substantial amounts of DNA are introduced into and deleted from the chromosome. These lateral transfers have effectively changed the ecological and pathogenic character of bacterial species. PMID- 10830952 TI - Oceanic Cd/P ratio and nutrient utilization in the glacial Southern Ocean AB - During glacial periods, low atmospheric carbon dioxide concentration has been associated with increased oceanic carbon uptake, particularly in the southern oceans. The mechanism involved remains unclear. Because ocean productivity is strongly influenced by nutrient levels, palaeo-oceanographic proxies have been applied to investigate nutrient utilization in surface water across glacial transitions. Here we show that present-day cadmium and phosphorus concentrations in the global oceans can be explained by a chemical fractionation during particle formation, whereby uptake of cadmium occurs in preference to uptake of phosphorus. This allows the reconstruction of past surface water phosphate concentrations from the cadmium/calcium ratio of planktonic foraminifera. Results from the Last Glacial Maximum show similar phosphate utilization in the subantarctic to that of today, but much smaller utilization in the polar Southern Ocean, in a model that is consistent with the expansion of glacial sea ice and which can reconcile all proxy records of polar nutrient utilization. By restricting communication between the ocean and atmosphere, sea ice expansion also provides a mechanism for reduced CO2 release by the Southern Ocean and lower glacial atmospheric CO2. PMID- 10830954 TI - Discovery of a comet by its Lyman-alpha emission AB - Several searches for near-Earth objects have recently been initiated, as a result of increased awareness of the hazard of impacts on the Earth. These programs mainly search for asteroids, so amateur astronomers can still contribute to the discovery of comets, especially out of the orbital plane of the Solar System. An ideal way to search for comets would be to use a spaceborne instrument capable of imaging the whole sky on a daily basis in a systematic and repeatable way. Such an instrument already exists on the solar observatory SOHO; it operates at the Lyman-alpha wavelength of neutral hydrogen, which is the main component of the emission cloud of a comet. Here we report the discovery, using archival data from this satellite, of a hitherto unnoticed comet which reached a perihelion of 1.546 a.u. on 26 June 1997. We derive the water production rate of the comet as a function of time and find that it increases after perihelion, like that of comet Halley. PMID- 10830953 TI - The DNA sequence of human chromosome 21. AB - Chromosome 21 is the smallest human autosome. An extra copy of chromosome 21 causes Down syndrome, the most frequent genetic cause of significant mental retardation, which affects up to 1 in 700 live births. Several anonymous loci for monogenic disorders and predispositions for common complex disorders have also been mapped to this chromosome, and loss of heterozygosity has been observed in regions associated with solid tumours. Here we report the sequence and gene catalogue of the long arm of chromosome 21. We have sequenced 33,546,361 base pairs (bp) of DNA with very high accuracy, the largest contig being 25,491,867 bp. Only three small clone gaps and seven sequencing gaps remain, comprising about 100 kilobases. Thus, we achieved 99.7% coverage of 21q. We also sequenced 281,116 bp from the short arm. The structural features identified include duplications that are probably involved in chromosomal abnormalities and repeat structures in the telomeric and pericentromeric regions. Analysis of the chromosome revealed 127 known genes, 98 predicted genes and 59 pseudogenes. PMID- 10830955 TI - Thermally fluctuating superconductors in two dimensions AB - In many two-dimensional superconducting systems, such as Josephson-junction arrays, granular superconducting films, and the high-temperature superconductors, it appears that the electrons bind into Cooper pairs below a pairing temperature (T(P)) that is well above the Kosterlitz-Thouless temperature (T(KT)) the temperature below which there is long-range superconducting order). The electron dynamics at temperatures between T(KT) and T(P) involve a complex interplay of thermal and quantum fluctuations, for which no quantitative theory exists. Here we report numerical results for this region, by exploiting its proximity to a T = 0 superconductor-insulator quantum phase transition. This quantum critical point need not be experimentally accessible for our results to apply. We characterize the static, thermodynamic properties by a single dimensionless parameter, gamma(T). Quantitative and universal results are obtained for the frequency dependence of the conductivity, which are dependent only upon gamma(T) and fundamental constants of nature. PMID- 10830956 TI - Optical microscopy using a single-molecule light source AB - Rapid progress in science on nanoscopic scales has promoted increasing interest in techniques of ultrahigh-resolution optical microscopy. The diffraction limit can be surpassed by illuminating an object in the near field through a sub wavelength aperture at the end of a sharp metallic probe. Proposed modifications of this technique involve replacing the physical aperture by a nanoscopic active light source. Advances in the spatial and spectral detection of individual fluorescent molecules, using near-field and far-field methods, suggest the possibility of using a single molecule as the illumination source. Here we present optical images taken with a single molecule as a point-like source of illumination, by combining fluorescence excitation spectroscopy with shear-force microscopy. Our single-molecule probe has potential for achieving molecular resolution in optical microscopy; it should also facilitate controlled studies of nanometre-scale phenomena (such as resonant energy transfer) with improved lateral and axial spatial resolution. PMID- 10830957 TI - Intercalation of alkylamines into an organic polymer crystal AB - Organic solid-state synthesis allows formation of products that are difficult or impossible to produce by conventional methods. This feature, and the high degree of reaction selectivity that can be achieved, is a direct result of the control over the relative orientation of the reactants afforded by the solid state. But as the successful development of 'topochemical reactions' requires the careful design of suitable reactant crystals, the range of both reactions and products amenable to this approach has been limited. However, recent advances in organic crystal engineering, particularly the rational design of complex solid architectures through supramolecular preorganization, have renewed interest in topochemical reactions. Previously, we have orientated muconate monomers--diene moieties with a carboxylate group on each end--using long-chain n-alkylammonium ions, such that the topochemical photopolymerization of the solid-state reactants produces layered crystals of stereoregular and high-molecular-mass polymers. Here we show that these polymer crystals are capable of repeated, reversible intercalation by conversion to the analogous poly(carboxylic acid), followed by transformation into a number of poly(alkylammonium muconate)s upon addition of the appropriate amine. Introduction of functional groups into these crystals may allow the design of organic solids for applications such as molecular recognition, separation and catalysis, thereby extending the range and practical utility of current intercalation compounds. PMID- 10830958 TI - Constraining the atmospheric N2O budget from intramolecular site preference in N2O isotopomers AB - Nitrous oxide (N2O) is an important trace gas in the atmosphere. It is an active greenhouse gas in the troposphere and it also controls ozone concentration in the stratosphere through nitric oxide production. One way to trace the geochemical cycle of N2O is by measuring the natural abundance of stable isotopes, namely 15N and 18O (refs 2-15). Here we report the intramolecular distribution of 15N within the linear NNO molecule, determined by measuring molecular and fragment ions of N2O on a modified mass spectrometer. This revealed a preference for 15N at the central N position, or alpha-site, within N2O isotopomers (isotope-containing molecules). Moreover, this preference varied significantly throughout the atmosphere. In the troposphere, low alpha-site preference indicates local emission of N2O from soils and fossil-fuel combustion, each with distinct isotopomer signatures, which then mixes with background N2O. In the stratosphere, on the other hand, loss of N2O is observed as enhanced alpha-site preference for 15N, due to fractionation during ultraviolet photolysis of N2O. We have constructed an atmospheric mass balance of N2O, incorporating isotopomer abundance, which shows that the intramolecular distribution of 15N is a parameter that has the potential to increase significantly the resolution with which sources and sinks of N2O can be identified and quantified in the atmosphere. PMID- 10830959 TI - Stimulated influences of Lake Agassiz on the climate of central North America 11,000 years ago AB - Eleven thousand years ago, large lakes existed in central and eastern North America along the margin of the Laurentide Ice Sheet. The large-scale North American climate at this time has been simulated with atmospheric general circulation models, but these relatively coarse global models do not resolve potentially important features of the mesoscale circulation that arise from interactions among the atmosphere, ice sheet, and proglacial lakes. Here we present simulations of the climate of central and eastern North America 11,000 years ago with a high-resolution, regional climate model nested within a general circulation model. The simulated climate is in general agreement with that inferred from palaeoecological evidence. Our experiments indicate that through mesoscale atmospheric feedbacks, the annual delivery of moisture to the Laurentide Ice Sheet was diminished at times of a large, cold Lake Agassiz relative to periods of lower lake stands. The resulting changes in the mass balance of the ice sheet may have contributed to fluctuations of the ice margin, thus affecting the routing of fresh water to the North Atlantic Ocean. A retreating ice margin during periods of high lake level may have opened an outlet for discharge of Lake Agassiz into the North Atlantic. A subsequent advance of the ice margin due to greater moisture delivery associated with a low lake level could have dammed the outlet, thereby reducing discharge to the North Atlantic. These variations may have been decisive in causing the Younger Dryas cold event. PMID- 10830960 TI - Mesozoic plate-motion history below the northeast Pacific Ocean from seismic images of the subducted Farallon slab AB - The high-resolution seismic imaging of subducted oceanic slabs has become a powerful tool for reconstructing palaeogeography. The images can now be interpreted quantitatively by comparison with models of the general circulation of the Earth's mantle. Here we use a three-dimensional spherical computer model of mantle convection to show that seismic images of the subducted Farallon plate provide strong evidence for a Mesozoic period of low-angle subduction under North America. Such a period of low-angle subduction has been invoked independently to explain Rocky Mountain uplift far inland from the plate boundary during the Laramide orogeny. The computer simulations also allow us to locate the largely unknown Kula-Farallon spreading plate boundary, the location of which is important for inferring the trajectories of 'suspect' terrain across the Pacific basin. PMID- 10830961 TI - Functional diversity governs ecosystem response to nutrient enrichment. AB - The relationship between species diversity and ecosystem functioning is a central topic in ecology today. Classical approaches to studying ecosystem responses to nutrient enrichment have considered linear food chains. To what extent ecosystem structure, that is, the network of species interactions, affects such responses is currently unknown. This severely limits our ability to predict which species or functional groups will benefit or suffer from nutrient enrichment and to understand the underlying mechanisms. Here our approach takes ecosystem complexity into account by considering functional diversity at each trophic level. We conducted a mesocosm experiment to test the effects of nutrient enrichment in a lake ecosystem. We developed a model of intermediate complexity, which separates trophic levels into functional groups according to size and diet. This model successfully predicted the experimental results, whereas linear food chain models did not. Our model shows the importance of functional diversity and indirect interactions in the response of ecosystems to perturbations, and indicates that new approaches are needed for the management of freshwater ecosystems subject to eutrophication. PMID- 10830962 TI - Adaptive plasticity in mate preference linked to differences in reproductive effort. AB - There is abundant evidence for the existence of marked mate preferences in natural populations, but the occurrence of within-population variation in mate preferences has received little attention and is often regarded as nonadaptive deviation from the optimal norm. Here we show experimentally that the preference of female collared flycatchers Ficedula albicollis for male forehead patch size, a sexually selected trait, varies with the time of breeding, an environmental factor with strong effects on reproductive success. Contrary to expectations based on time-constrained choice models, only late-breeding females prefer males with a large patch size. The variation in mate preference matches a seasonal change in female reproductive success: long-term data reveal a positive relationship between female reproductive success and male patch size exclusively in late breeders. In addition, female reproductive effort, as assessed by clutch size, appears to be adjusted relative to both timing of breeding and male phenotype. We conclude that not only can mate preferences display adaptive plasticity within populations, but this plasticity can also be linked to differences in reproductive investment. PMID- 10830963 TI - Cross-modal and cross-temporal association in neurons of frontal cortex. AB - The prefrontal cortex is essential for the temporal integration of sensory information in behavioural and linguistic sequences. Such information is commonly encoded in more than one sense modality, notably sight and sound. Connections from sensory cortices to the prefrontal cortex support its integrative function. Here we present the first evidence that prefrontal cortex cells associate visual and auditory stimuli across time. We gave monkeys the task of remembering a tone of a certain pitch for 10 s and then choosing the colour associated with it. In this task, prefrontal cortex cells responded selectively to tones, and most of them also responded to colours according to the task rule. Thus, their reaction to a tone was correlated with their subsequent reaction to the associated colour. This correlation faltered in trials ending in behavioural error. We conclude that prefrontal cortex neurons are part of integrative networks that represent behaviourally meaningful cross-modal associations. The orderly and timely activation of neurons in such networks is crucial for the temporal transfer of information in the structuring of behaviour, reasoning and language. PMID- 10830964 TI - Phenotypic suppression of empty spiracles is prevented by buttonhead. AB - Unlike the trunk segments, the anterior head segments of Drosophila are formed in the absence of pair-rule and HOX-cluster gene expression, by the activities of the gap-like genes orthodenticle (otd), empty spiracles (ems) and buttonhead (btd). The products of these genes are transcription factors, but only EMS has a HOX-like homeodomain. Indeed, ems can confer identity to trunk segments when other HOX-cluster gene activities are absent. In trunk segments of wild-type embryos, however, ems activity is prevented by phenotypic suppression, in which more posterior HOX-cluster genes inactivate the more anterior without affecting transcription or translation. ems is suppressed by all other Hox-cluster genes and so is placed at the bottom of their hierarchy. Here we show that misexpression of EMS in the head transforms segment identity in a btd-dependent manner, that misexpression of BTD in the trunk causes ems-dependent structures to develop, and that EMS and BTD interact in vitro. The data indicate that this interaction may allow ems to escape from the bottom of the HOX-cluster gene hierarchy and cause a dominant switch of homeotic prevalence in the anterior posterior direction. PMID- 10830965 TI - Blockade of RAGE-amphoterin signalling suppresses tumour growth and metastases. AB - The receptor for advanced glycation end products (RAGE), a multi-ligand member of the immunoglobulin superfamily of cell surface molecules, interacts with distinct molecules implicated in homeostasis, development and inflammation, and certain diseases such as diabetes and Alzheimer's disease. Engagement of RAGE by a ligand triggers activation of key cell signalling pathways, such as p21ras, MAP kinases, NF-kappaB and cdc42/rac, thereby reprogramming cellular properties. RAGE is a central cell surface receptor for amphoterin, a polypeptide linked to outgrowth of cultured cortical neurons derived from developing brain. Indeed, the co localization of RAGE and amphoterin at the leading edge of advancing neurites indicated their potential contribution to cellular migration, and in pathologies such as tumour invasion. Here we demonstrate that blockade of RAGE-amphoterin decreased growth and metastases of both implanted tumours and tumours developing spontaneously in susceptible mice. Inhibition of the RAGE-amphoterin interaction suppressed activation of p44/p42, p38 and SAP/JNK MAP kinases; molecular effector mechanisms importantly linked to tumour proliferation, invasion and expression of matrix metalloproteinases. PMID- 10830966 TI - Neurotoxicity induces cleavage of p35 to p25 by calpain. AB - Cyclin-dependent kinase 5 (cdk5) and its neuron-specific activator p35 are required for neurite outgrowth and cortical lamination. Proteolytic cleavage of p35 produces p25, which accumulates in the brains of patients with Alzheimer's disease. Conversion of p35 to p25 causes prolonged activation and mislocalization of cdk5. Consequently, the p25/cdk5 kinase hyperphosphorylates tau, disrupts the cytoskeleton and promotes the death (apoptosis) of primary neurons. Here we describe the mechanism of conversion of p35 to p25. In cultured primary cortical neurons, excitotoxins, hypoxic stress and calcium influx induce the production of p25. In fresh brain lysates, addition of calcium can stimulate cleavage of p35 to p25. Specific inhibitors of calpain, a calcium-dependent cysteine protease, effectively inhibit the calcium-induced cleavage of p35. In vitro, calpain directly cleaves p35 to release a fragment with relative molecular mass 25,000. The sequence of the calpain cleavage product corresponds precisely to that of p25. Application of the amyloid beta-peptide A beta(1-42) induces the conversion of p35 to p25 in primary cortical neurons. Furthermore, inhibition of cdk5 or calpain activity reduces cell death in A beta-treated cortical neurons. These observations indicate that cleavage of p35 to p25 by calpain may be involved in the pathogenesis of Alzheimer's disease. PMID- 10830968 TI - Translocation step size and mechanism of the RecBC DNA helicase. AB - DNA helicases are ubiquitous enzymes that unwind double-stranded DNA. They are a diverse group of proteins that move in a linear fashion along a one-dimensional polymer lattice--DNA--by using a mechanism that couples nucleoside triphosphate hydrolysis to both translocation and double-stranded DNA unwinding to produce separate strands of DNA. The RecBC enzyme is a processive DNA helicase that functions in homologous recombination in Escherichia coli; it unwinds up to 6,250 base pairs per binding event and hydrolyses slightly more than one ATP molecule per base pair unwound. Here we show, by using a series of gapped oligonucleotide substrates, that this enzyme translocates along only one strand of duplex DNA in the 3'-->5' direction. The translocating enzyme will traverse, or 'step' across, single-stranded DNA gaps in defined steps that are 23 (+/-2) nucleotides in length. This step is much larger than the amount of double-stranded DNA that can be unwound using the free energy derived from hydrolysis of one molecule of ATP, implying that translocation and DNA unwinding are separate events. We propose that the RecBC enzyme both translocates and unwinds by a quantized, two-step, inchworm-like mechanism that may have parallels for translocation by other linear motor proteins. PMID- 10830967 TI - LAG-3 is a putative transcriptional activator in the C. elegans Notch pathway. AB - Notch signalling controls growth, differentiation and patterning during normal animal development; in humans, aberrant Notch signalling has been implicated in cancer and stroke. The mechanism of Notch signalling is thought to require cleavage of the receptor in response to ligand binding, movement of the receptor's intracellular domain to the nucleus, and binding of that intracellular domain to a CSL (for CBF1, Suppressor of Hairless, LAG-1) protein. Here we identify LAG-3, a glutamine-rich protein that forms a ternary complex together with the LAG-1 DNA-binding protein and the receptor's intracellular domain. Receptors with mutant ankyrin repeats that abrogate signal transduction are incapable of complex formation both in yeast and in vitro. Using RNA interference, we find that LAG-3 activity is crucial in Caenorhabditis elegans for both GLP-1 and LIN-12 signalling. LAG-3 is a potent transcriptional activator in yeast, and a Myc-tagged LAG-3 is predominantly nuclear in C. elegans. We propose that GLP-1 and LIN-12 promote signalling by recruiting LAG-3 to target promoters, where it functions as a transcriptional activator. PMID- 10830969 TI - The crystal structure of the photoprotein aequorin at 2.3 A resolution. AB - Aequorin is a calcium-sensitive photoprotein originally obtained from the jellyfish Aequorea aequorea. Because it has a high sensitivity to calcium ions and is biologically harmless, aequorin is widely used as a probe to monitor intracellular levels of free calcium. The aequorin molecule contains four helix loop-helix 'EF-hand' domains, of which three can bind calcium. The molecule also contains coelenterazine as its chromophoric ligand. When calcium is added, the protein complex decomposes into apoaequorin, coelenteramide and CO2, accompanied by the emission of light. Apoaequorin can be regenerated into active aequorin in the absence of calcium by incubation with coelenterazine, oxygen and a thiol agent. Cloning and expression of the complementary DNA for aequorin were first reported in 1985 (refs 2, 6), and growth of crystals of the recombinant protein has been described; however, techniques have only recently been developed to prepare recombinant aequorin of the highest purity, permitting a full crystallographic study. Here we report the structure of recombinant aequorin determined by X-ray crystallography. Aequorin is found to be a globular molecule containing a hydrophobic core cavity that accommodates the ligand coelenterazine 2-hydroperoxide. The structure shows protein components stabilizing the peroxide and suggests a mechanism by which calcium activation may occur. PMID- 10830970 TI - Effect of intrathecal morphine and electro-acupuncture on cellular immune function of rats and increment of mu-opioid receptor mRNA expression in PAG following intrathecal morphine. AB - The present study was to investigate the dynamic changes of cellular immune function of rats with intrathecal injection of (ith) morphine and the regulatory effect of electroacupuncture(EA) stimulation on "Zusanli" (St.36) and "Lanwei" (Extra 37) points. The results showed that ConA-induced rat spleen lymphocyte proliferation was significantly decreased on 2h, 4h, 8h, 12h, 24h, 48h after ith morphine(40microg/50microL). The proliferative response was recovered to nearly normal on 72h. EA on corresponding periods could prevent the decrease of lymphocyte proliferative response of rats induced by ith morphine. The same tendency was observed on the induction of IL-2 production. Further study continued to explore the mechanism of the potentiating effect of mu-opioid receptor in periaqueductal gray (PAG) and hippocampus on the immunosuppression induced by ith morphine at molecular level with in situ hybridization histochemistry technique. The results showed that ith morphine could increase the expression of mu-opioid receptor mRNA. PMID- 10830971 TI - Osteoreflectory treatment of alcohol abstinence syndrome and craving for alcohol in patients with alcoholism. AB - Osteoreflexotherapy is used alone as a treatment used for alcohol abstinence syndrome and for alcohol craving by intraosseal stimulation of the processus styloideus ulnae of the patient's left and right hands as well as the processus spinosus of the seventh cervical vertebra and the manubrium sterni osteoreceptors. This is done by intraosseal injection of 0.5 to 1.0 ml of 0.9% NaCl solution during a period of 3 to 5 seconds. Craving for alcohol and depressed mood, strongly manifested Alcohol Abstinence Syndrome (AAS) symptoms before osteoreflexotherapy, were reduced in a most convenient and fast manner under the influence of two sessions of osteoreceptive stimulation. The withdrawal symptoms caused by alcohol abstinence decreased markedly during the first two hours after the first osteoreflexotherapy treatment, continued to decrease in the next 24 hours and by the time the second osteoreflexotherapy session was given, the withdrawal symptoms completely disappeared in 72 hours. The most slowly and least reduced AAS symptoms were asthenia and disturbances of postular equilibrium. Based on clinical observations, it is speculated that osteoreceptive stimulations destroy ethanol dependent functional systems and restore the neurophysiological and neuromediatorial integration of the brain in alcoholismpatients. Primarily because of these two cited factors, the patient can be freed of the craving for alcohol for several years, and he or she also does not suffer from depression. PMID- 10830972 TI - Effect of morphine and electro-acupuncture (EA) on apoptosis of thymocytes. AB - The influence of morphine and EA on the apoptosis of thymocytes were studied to investigate the posibility of its involvement in the mechanism of morphine induced immunosuppression and the regulatory effect of EA on it. 1h after injecting 50 mg/kg morphine subcutaneously into 3-wk old Balb/c mice continually twice a day for 5 days, thymus was collected and the apoptotic cell was detected by a method of terminal deoxynucleotidyl transferase-meditaed dUTP nick end labeling(TUNEL). The results showed that morphine significantly enhanced the percentage of TUNEL positive cells inside thymus with an appearing of apoptotic DNA ladder after 24 h incubation. Treating mice with EA of "Zusanli(St.36)" and "Lanwei(Ext.33)" for 1h after morphine administration decreased the percentage of TUNEL positive cells. EA also showed an regulatory effect on the increased the expression of CPP32 and decreased the expression of Bcl-2 by morphine. The significant enhancement of hypothalamic CRF and plasma ACTH level by morphine and the antagonize effect of EA on it suggested a possible role of Hypothalamus pituitary-adrenal (HPA) axis played in the apoptosis of thymocytes by morphine and the regulatory effect of EA. PMID- 10830973 TI - Effect of high or low frequency electroacupuncture on the cellular activity of catecholaminergic neurons in the brain stem. AB - Although the opioid system plays a pivotal role in the analgesic effect of electroacupuncture (EA), it has been suggested that other peptidergic systems also may be involved in the therapeutic effect of EA. Among several peptides for EA-induced analgesia, catecholamine (CA) is associated with the descending pain inhibitory system. We evaluated whether the different frequencies of EA modified the cellular activity of central CA synthesizing neurons using double labeling immunohistochemistry between Fos-like immunoreactive (FLI) neurons and dopamine beta-hydroxylase (DBH)/tyrosine hydroxylase (TH)-positive neurons. We observed that different frequencies of EA increased the number of FLI neurons in catecholaminergic neurons, such as the dorsal raphe (DR), hypothalamic arcuate nucleus (Arc), locus coeruleus (LC), A5 noradrenaline cells (A5), and A7 noradrenaline cells (A7). In addition, different frequencies of EA significantly increased the ratio of colocalization between FLI neurons and TH positive neurons in DR, LC and Arc. Only low frequency EA increased the neuronal activity in Arc. The ratio of double labeling between FLI and DBH positive neurons was also elevated at both LC and A5. These data demonstrate that different frequencies of EA increase the cellular activity of central CA synthesizing neurons, suggesting that the CA system plays an important role in EA-induced analgesia. PMID- 10830974 TI - Sleep patterns and sleep disruptions in school-age children. AB - This study assessed the sleep patterns, sleep disruptions, and sleepiness of school-age children. Sleep patterns of 140 children (72 boys and 68 girls; 2nd-, 4th-, and 6th-grade students) were evaluated with activity monitors (actigraphs). In addition, the children and their parents completed complementary sleep questionnaires and daily reports. The findings reflected significant age differences, indicating that older children have more delayed sleep onset times and increased reported daytime sleepiness. Girls were found to spend more time in sleep and to have an increased percentage of motionless sleep. Fragmented sleep was found in 18% of the children. No age differences were found in any of the sleep quality measures. Scores on objective sleep measures were associated with subjective reports of sleepiness. Family stress, parental age, and parental education were related to the child's sleep-wake measures. PMID- 10830975 TI - Sociable and prosocial dimensions of social competence in Chinese children: common and unique contributions to social, academic, and psychological adjustment. AB - A sample of 6th-grade children (N = 470), initially aged 12 years, in Shanghai, People's Republic of China, participated in this longitudinal study. Data on children's sociability and prosocial orientation were collected from peer assessments. Information on indexes of adjustment was obtained from multiple sources. The results indicated that sociable and prosocial functions were 2 overlapping, but distinct, dimensions of social competence and had common as well as unique contributions to the prediction of adjustment in specific areas. Whereas prosocial orientation was mainly predictive of social and school adjustment and externalizing problems, sociability uniquely contributed to the prediction of internalizing problems and emotional adjustment. Finally, prosocial orientation and sociability might moderate the development of adjustment difficulties. PMID- 10830976 TI - How Latino American and European American adolescents discuss conflicts, sexuality, and AIDS with their mothers. AB - The authors examined how the structure of mother-adolescent conversations differs by ethnic group, age, and dyadic and individual factors. Mother-adolescent dyads of European or Latino descent participated in conversations and reported on their relationship and AIDS knowledge. Latina American mothers dominated conversations more than European American mothers, independent of socioeconomic status. Mothers dominated conversations about sexuality and AIDS more than conversations about conflicts. Mothers of older adolescents reacted more negatively, and older adolescents reported less satisfaction, less openness, and more sexual discussions with persons other than their mothers. Latino American adolescents whose mothers dominated conversations more reported fewer sexual discussions. Latina American mothers who dominated conversations more reported more openness and satisfaction. When mothers dominated conversations more, adolescents had lower AIDS knowledge. PMID- 10830977 TI - Friendship and social competence in a sample of preschool children attending Head Start. AB - Relations between friendship (operationalized as reciprocated or nonreciprocated sociometric choices) and social competence were studied for children (mostly African American) attending Head Start. Initial analyses showed that children with reciprocated friends had higher social competence scores than children without reciprocated friends. Correlations suggested that the number of reciprocated friendships was associated with the social competence indicators studied here. Beyond the cost of having no reciprocated friends, having nonreciprocated friendships was not a liability. Cross-time analyses suggested differing patterns of relations for boys and girls. Having versus not having a reciprocated friend was unstable across time, because there was a trend toward participating in reciprocated friendships from 3 to 4 years of age (most older children had at least 1 reciprocated friend). For girls there was a positive relation between the number of reciprocated friendships at Time 1 and at Time 2. No benefit (in terms of social competence) was found for children making the transition from 1 classroom to the next with a friend. PMID- 10830978 TI - Shared caregiving: comparisons between home and child-care settings. AB - The experiences of 84 German toddlers (12-24 months old) who were either enrolled or not enrolled in child care were described with observational checklists from the time they woke up until they went to bed. The total amount of care experienced over the course of a weekday by 35 pairs of toddlers (1 member of each pair in child care, 1 member not) did not differ according to whether the toddlers spent time in child care. Although the child-care toddlers received lower levels of care from care providers in the centers, their mothers engaged them in more social interactions during nonworking hours than did the mothers of home-only toddlers, which suggests that families using child care provided different patterns of care than families not using child care. Child-care toddlers experienced high levels of emotional support at home, although they experienced less prompt responses to their distress signals. Mothers' ages were unrelated to the amounts of time toddlers spent with them, but older mothers initiated more proximity. PMID- 10830979 TI - Children's use of mental state information in selecting persuasive arguments. AB - Whether and when children use information about others' mental states to invent or select persuasive strategies were examined. In Study 1, preschoolers, 3rd graders, and 6th-graders (ns = 11, 12, and 16, respectively; 17 girls) were told about story characters' persuading parents to buy pets or toys. Children were either given or not given information about story parents' beliefs and asked to invent or select appropriate arguments. Older children, but not preschoolers, used belief information to select arguments. Results were replicated in Study 2 (16 kindergartners, 16 3rd-graders; 19 girls). In Study 3, kindergartners and 1st graders (N = 16; 6 girls) reasoned well on false-belief tasks but not on persuasion tasks, suggesting that failure to consider mental states in persuasion was not due to lack of a belief concept. Findings suggest that mental state understanding may continue to develop after the preschool years; methodological qualifications are also considered. PMID- 10830980 TI - What parents know, how they know it, and several forms of adolescent adjustment: further support for a reinterpretation of monitoring. AB - Parental monitoring has been conceptualized as tracking and surveillance but operationalized as knowledge of daily activities. This study tested the tracking and surveillance explanation of why parental knowledge is linked to better adolescent adjustment. Participants were 1,186 14-year-olds in central Sweden and their parents. The results supported and extended a reinterpretation of parental monitoring (H. Stattin & M. Kerr, in press). Across sex and informant, high parental knowledge was linked to multiple measures of good adjustment. But children's spontaneous disclosure of information explained more of these relations than parents' tracking and surveillance efforts did. Parents' control efforts were related to good adjustment only after the child's feelings of being controlled, which were linked to poor adjustment, were partialed out. The findings suggest that parents' tracking and surveillance efforts are not as effective as previously thought. PMID- 10830981 TI - Gender, affiliation, assertion, and the interactive context of parent-child play. AB - Ninety-eight young U.S. children (mean age = 48 months) with either European, Latin American, or multiple ethnic backgrounds were videotaped with their mothers and their fathers on separate occasions in their families' homes. Parent-child pairs played for 8 min each with a feminine-stereotyped toy set (foods and plates) and a masculine-stereotyped toy set (track and cars). Levels of affiliation (engaging vs. distancing) and assertion (direct vs. nondirect) were rated on 7-point scales every 5 s from the videotapes for both parent and child. Overall, the play activity accounted for a large proportion of the variance in parents' and children's mean affiliation and assertion ratings. Some hypothesized gender-related differences in behavior were also observed. In addition, exploratory analyses revealed some differences between the different ethnic groups. The results highlight the importance of role modeling and activity settings in the socialization and social construction of gender. PMID- 10830982 TI - Where's the ball? Two- and three-year-olds reason about unseen events. AB - Children 2, 2 1/2, and 3 years of age engaged in a search task in which they opened 1 of 4 doors in an occluder to retrieve a ball that had been rolled behind the occluder. The correct door was determined by a partially visible wall placed behind the occluder that stopped the motion of the unseen ball. Only the oldest group of children was able to reliably choose the correct door. All children were able to retrieve a toy that had been hidden in the same apparatus if the toy was hidden from the front by opening a door. Analysis of the younger children's errors indicated that they did not search randomly but instead used a variety of strategies. The results are consistent with the Piagetian view that the ability to use representations to guide action develops slowly over the first years of life. PMID- 10830983 TI - Maternal illusory control predicts socialization strategies and toddler compliance. AB - This study examined the relation between mothers' perception of their capacity for controlling infant crying and a later measure of parent-toddler social interaction-compliance with parental requests. Fifty-seven mothers participated in (a) a laboratory task when their children were 5 months old to assess mothers' perception of control and (b) a toy-cleanup task when their children were 24 months old to assess toddler compliance. Perception-of-control scores on the laboratory task reflected accuracy of perception and ranged from accurate to greatly overestimating control, with an overestimation referred to as an illusion of control. Mothers were categorized into three illusion-of-control groups: low, moderate, and high. Toddlers of mothers in the low and high illusion-of-control groups were more likely to be categorized as highly defiant than were toddlers of mothers in the moderate-illusion group. Mothers with high illusory control were most likely to use a high power-assertion strategy (negative control), and when negotiating, their toddlers' expression of autonomy was most likely to escalate into defiance. PMID- 10830984 TI - Cancer registration in the United States. PMID- 10830985 TI - Usefulness of serum carboxy-terminal telopeptide of type I collagen (ICTP) as a marker of bone metastasis from lung cancer. AB - BACKGROUND: Serum pyridinoline cross-linked carboxy-terminal telopeptide of type I collagen (ICTP) is a metabolite of type I collagen comprising 90% or more of organic substances in bone. Its usefulness as a marker of bone metastasis from malignant tumors is expected. METHOD: We measured ICTP to evaluate its clinical usefulness for diagnosis of bone metastasis in 140 patients with lung cancer. For comparison, serum carcinoembryonic antigen (CEA), cytokeratin 19 fragment (CYFRA 21-1), gastrin-releasing peptide precursor (ProGRP), alkaline phosphatase and calcium were simultaneously measured. ICTP was measured by double-antibody radioimmunoassay. RESULTS: ICTP was significantly higher in patients with bone metastasis from lung cancer than in the group without bone metastasis, patients with other pulmonary diseases or healthy control subjects and showed excellent sensitivity and specificity, indicating that this marker is highly useful for complementary diagnosis of bone metastasis from lung cancer. Moreover, the survival duration was significantly shorter in the ICTP-positive group than in the ICTP-negative group, suggesting that ICTP can be a prognostic factor in lung cancer. CONCLUSION: It is suggested that measurement of ICTP is worthwhile as a serological diagnostic method of bone metastasis from lung cancer. Moreover, since repeated measurements are possible, this measure was considered very helpful in complementary diagnosis of bone metastasis and also as a standard to determine the timing of examinations such as bone scintigraphy. PMID- 10830986 TI - Reduction of late complications after irregularly shaped four-field whole pelvic radiotherapy using computed tomographic simulation compared with parallel-opposed whole pelvic radiotherapy. AB - BACKGROUND: Tumor control and late complication rates of irregularly shaped four field whole pelvic radiotherapy using CT simulation were compared with those of whole pelvic radiotherapy using parallel-opposed fields in a non-randomized study. METHODS: From 1986 to 1996, 74 patients who underwent surgery for clinical stage I, II or III squamous or adenosquamous cell carcinoma of the uterine cervix were treated with postoperative radiotherapy consisting of 50 Gy in 25 fractions in 6 weeks. Thirty-four patients were treated with an irregularly shaped four field technique following computed tomography (CT) simulation using beam's eye view and three-dimensional treatment planning and lead blocks. Forty patients received the conventional two-field technique, with CT simulation in 13 patients and X-ray simulation in 27 patients. There was no significant difference in patients' characteristics between the two groups. RESULTS: There was no statistical difference in survival, relapse-free survival or pelvic control rate between the two-field and irregularly shaped four-field groups with a mean follow up period of 60 months. The actual 5-year pelvic control rate was 94% for the two field technique and 100% for the irregularly shaped four-field technique. The incidence of grade II-III bowel complications in the irregularly shaped technique group (2.9%, 1/34) was significantly lower than that in the two-field technique group (17.5%, 7/40) (p < 0.05). The actual 5-year complication rates of grade II leg edema were 28.6 and 3.1% for the two-field technique and irregularly shaped four-field technique groups, respectively (p = 0.0123). CONCLUSIONS: Irregularly shaped four-field post-operative pelvic radiotherapy using CT simulation appears to be as effective as parallel-opposed whole pelvic radiotherapy with a lower incidence of bowel complication and chronic leg edema. PMID- 10830987 TI - Occult breast cancer presenting axillary nodal metastasis: a case report. AB - We report a case of a 42-year-old female with occult breast cancer presenting axillary nodal metastasis. She complained of a swelling of the right axillary lymph node, but no breast mass was palpable. Biopsy of the lymph node was performed and histological examination showed a metastatic ductal carcinoma with papillotubular formation. Estrogen receptor of the lymph node was positive. No pathological findings were obtained by mammography and ultrasonography and systemic examinations revealed no extramammary primary lesion. All these data suggested an occult carcinoma of the breast and modified radical mastectomy was performed. Pathological findings of the removed specimen failed to find the primary breast cancer lesion. The patient has been treated with hormonal therapy and she is well without evidence of disease 5 years after surgery. PMID- 10830988 TI - Bilateral male breast cancer and prostate cancer: a case report. AB - Male breast cancer, consisting only 1% of all breast cancers, is occasionally associated with other primary malignancies, especially in patients with familial breast cancer history. Sporadic male breast cancers with another primary tumor are extremely rare. We report a 67-year-old male with asynchronous bilateral breast cancer and prostate cancer without familial breast cancer history. PMID- 10830989 TI - A surgical case of solitary plasmacytoma of rib origin with biclonal gammopathy. AB - Localized solitary plasmacytoma of the bone (SPB) is a rare disease and is characterized by only one or two isolated bone lesions with no evidence of disease dissemination. A previously healthy 44-year-old male was admitted for evaluation of an abnormal radiographic shadow in the left middle lung field with symptoms of left back pain. Radiological evaluation revealed a peripheral opacity in the left chest wall, which was highly suspected to be a chest wall tumor. CT guided transcutaneous needle biopsy of the tumor was performed and the specimens showed a monomorphous population of mature plasma cells. The bone marrow biopsy findings revealed no evidence of myeloma and bone scanning revealed only abnormal accumulation in the left seventh rib. He had mild M-proteins in a urine sample and Bence-Jones protein was detected. Immunoelectrophoresis revealed mild biclonal gammopathy of Bence-Jones protein of both the kappa and lambda light chain types. Under a diagnosis of solitary bone plasmacytoma, preoperative radiation therapy with doses of 40 Gy for the tumor was performed. He underwent complete en bloc resection of the chest wall, including one-third of the left sixth and seventh ribs, the intercostal muscle and the parietal pleura. The protein abnormalities in the urine sample disappeared following surgical resection. Adjuvant chemotherapy using melphalan and prednisolone was performed. He is doing well without evidence of tumor recurrence 2 years following his initial diagnosis. PMID- 10830990 TI - Inflammatory pseudotumor of the spleen: a case report. AB - We report on an inflammatory pseudotumor of the spleen. A 72-year-old woman visited our hospital complaining of nausea. Physical examination and laboratory investigations were unremarkable. Ultrasonography, computed tomography, magnetic resonance imaging and angiography showed a hypovascular splenic mass measuring about 5 cm in diameter with a calcification in the center of the lesion. Splenectomy was performed. The removed spleen, weighing 145 g, contained a tan white, circumscribed mass, measuring 6.2 x 5.5 x 5.3 cm. Histologically, the splenic mass was composed of an admixture of inflammatory cellular elements, predominantly plasma cells and lymphocytes with hyalinization, fibrosis, lymph follicles and multinuclear giant cells, suggestive of a inflammatory pseudotumor. The patient is currently alive and asymptomatic, 24 months after surgery. Inflammatory pseudotumors of the spleen are extremely rare and only 39 cases have been reported in the medical literature. PMID- 10830991 TI - A novel non-pathogenetic polymorphism of the APC gene in a patient with familial adenomatous polyposis coli. AB - DISORDER: Familial adenomatous polyposis coli. ETHNICITY OF PATIENT: Japanese. GENE: APC. GENBANK ACCESSION NUMBER: M 74088. CHROMOSOMAL ASSIGNMENT: 5q21. TYPE OF DNA VARIANT: A germline missense mutation. A germline nonsense mutation. MUTATION: CGG (Arg, wild type) to TGG (Trp) substitution at codon 88 in exon 3 of the APC gene. CGA (Arg, wild type) to TGA (term.) at codon 213 in exon 5 of the APC gene. ALLELIC FREQUENCY: <0.014 (missense mutation, TGG at codon 88). METHOD OF MUTATION DETECTION: PCR-SSCP/direct sequencing. PMID- 10830992 TI - An international meeting in Beijing. PMID- 10830993 TI - What is the ideal HER2 test for Herceptin therapy? PMID- 10830994 TI - Stomach cancer mortality by prefectures in Japan. PMID- 10830995 TI - How relevant are pollen and mold spore counts to clinical practice? PMID- 10830996 TI - The natural history of adult asthma: what do we know? PMID- 10830997 TI - Advances in immuno-ophthalmology. PMID- 10830998 TI - Care paths for acute asthma: an idea whose time has come. PMID- 10830999 TI - The use of newer asthma and allergy medications during pregnancy. The American College of Obstetricians and Gynecologists (ACOG) and The American College of Allergy, Asthma and Immunology (ACAAI). PMID- 10831000 TI - Interpreting atmospheric pollen counts for use in clinical allergy: spatial variability. AB - BACKGROUND: Atmospheric pollen counts are ubiquitous data, however, few guidelines exist for interpreting them. A fundamental problem involves appreciating spatial variability in atmospheric pollen concentrations, for example, differences in pollen conditions between neighborhoods within a metropolitan area. The significance of this problem is perhaps most manifest in clinical allergy where pollen counts from a single rooftop pollen sampler are often applied to an entire city. OBJECTIVES: This review will consider experimental data concerning spatial variability in atmospheric pollen concentrations. Several guidelines for interpreting pollen counts with respect to this spatial variability will be presented. DATA SOURCES: This review will consider three independent bodies of literature concerning spatial variability in atmospheric pollen concentrations: (1) controlled-release experiments, (2) the paleoecology literature, and (3) atmospheric surveys. RESULTS: Four principal results emerged. First, pollen concentrations in the atmosphere appear to be inherently heterogeneous. Differences in pollen recovery between adjacent points increase in a predictable manner as a function of distance. Second, spatial variability decreases as the distance between pollen sources and destination regions increases. Pollen recovery across a city is most similar when pollen is transported from distant source regions. Third, the size of the source area represented by a pollen sampler depends on the distance between the sampler and the nearest vegetation. Samplers located near vegetation reflect small source areas; source areas expand considerably when the distance to the nearest vegetation increases. Last, individual pollen types travel characteristic distances from their source. Heavy pollen types collected at a particular point reflect a smaller source areas than lighter types which represent much larger source areas. CONCLUSIONS: These conclusions allow allergists to interpret their atmospheric pollen counts with greater sophistication than was previously possible. Mathematical equations presented in this review permit rough calculations concerning source areas. PMID- 10831001 TI - Pulmonary vasculitis presenting as atypical asthma. PMID- 10831002 TI - Natural history of asthma: a 23-year followup of college students. AB - OBJECTIVE: A 23-year followup study of 1,601 college students who were initially evaluated for evidence of asthma and allergic rhinitis by direct interviews, physical examination, laboratory tests, and repeated questionnaires was conducted. METHODS AND RESULTS: The individuals were located through addresses from the alumnae office. Of these, 1,021 (64%) returned their completed questionnaires and these included 738 (72%) who had allergy skin tests as freshmen. A similar proportion of our total population of 1,021 were skin tested as freshmen compared with the original study population of 1,836 in the freshmen year (72% versus 68%). This difference was not statistically significant. Among these 738 alumnae, with a mean age of 40 years, there were a total of 84 with a history of asthma. At the time of the 23-year followup, 44 (52%) were considered to have active asthma and 40 (48%) were symptom free. A majority (85%) of those with inactive asthma were symptom free for 5 years or longer. Of those with active asthma, 50% felt they were improved, 39% unchanged, 9% felt worse, and 2% were unknown. Atopy was a non-prognostic indicator of asthma outcome as determined by scratch skin testing as college freshmen. New asthma occurred in 36 (5.2%) of those at risk to develop new asthma in this 23-year period for a rate of 0.23% per year. CONCLUSION: This 23-year followup study demonstrates that the cumulative prevalence of asthma continues to increase as the individuals become older. The asthma symptoms were no longer present or were improved in about three fourths of the asthma subjects in this 40-year-old age group. Of the remainder, most symptoms were unchanged and a small number felt worse. PMID- 10831003 TI - Olopatadine inhibits TNFalpha release from human conjunctival mast cells. AB - BACKGROUND: Tumor necrosis factor-alpha (TNFalpha) release likely plays a crucial role in allergic ocular inflammation via increasing ICAM-1 on epithelial cells and triggering other proinflammatory events. The immediate and prolonged release of TNFalpha from human conjunctival mast cells in response to allergen challenge is potentially an important target for therapeutic intervention, yet the effect of ocular anti-allergic agents on this process has not been examined. Olopatadine (Patanol) is a clinically effective dual-action ophthalmic anti-allergic agent that has been shown to inhibit mast cell histamine, tryptase, and PGD2 release in vitro and promote decreased H1 receptor binding activity in vitro and functional H1 receptor antagonism in vivo. OBJECTIVE: To investigate the effect of olopatadine on TNFalpha release from anti-IgE antibody challenged purified human conjunctival mast cells. METHODS: Human conjunctival mast cells were purified (>95%) from cadaveric tissues using a procedure combining enzymatic digestion and Percoll gradient centrifugation. These cells were incubated with olopatadine for 30 minutes then challenged with anti-IgE antibody for 90 minutes. Supernatants were analyzed for TNFalpha. RESULTS: Purified human conjunctival mast cells responded to anti-IgE antibody challenge with TNFalpha release in a concentration dependent manner (optimum concentration was 10 microg/mL). Olopatadine pre incubation resulted in a dose-dependent decrease in anti-IgE antibody mediated TNFalpha release (IC50 = 13.1 microM). At a concentration of 3 mM olopatadine reduced TNFalpha release to the level of unchallenged controls. CONCLUSION: Olopatadine inhibited anti-IgE antibody-mediated release of TNFalpha from human conjunctival mast cells. This effect could contribute to the long duration of anti-allergic activity reported for the drug. PMID- 10831004 TI - Improved outcomes for hospitalized asthmatic children using a clinical pathway. AB - BACKGROUND: Although asthma clinical pathways are used with increasing frequency, few controlled studies have evaluated the clinical and cost effectiveness of these pathways. OBJECTIVE: To evaluate the effect of an inpatient asthma clinical pathway on cost and quality of care for children with asthma. METHODS: One hundred forty-nine children were treated for status asthmaticus using an asthma clinical pathway in a children's hospital between September and December 1997. Thirty-four of 149 children treated with the clinical pathway were randomly selected. A retrospective cohort control group of non-pathway patients (N = 34) was matched with each pathway patient by age, race, gender, co-morbidities, asthma severity score, ICU admission, and time of year admitted. Differences between the two groups in length of stay, total costs, readmission rate, inpatient management, and discharge medications were compared. RESULTS: Length of stay was significantly lower in the clinical pathway group compared with the control group (36 hours versus 71 hours, P < .001) and total costs decreased significantly ($1685 versus $2829, P < .001) as a result of the pathway. Asthmatic children on the clinical pathway were significantly more likely than the control group to complete asthma teaching while hospitalized (65% versus 18%, P < .001), to be discharged with a prescription for a controller medication (88% versus 53%, P < .01), and to have a peak flow meter (57% versus 23%, P < .05) and a spacer device (100% versus 71%, P < .001) for home use. CONCLUSION: Implementation of this inpatient clinical pathway led to a decrease in length of stay and a reduction in total cost while improving quality of care for hospitalized asthmatic children. PMID- 10831005 TI - Fexofenadine HCl is safe and effective for treatment of chronic idiopathic urticaria. AB - BACKGROUND: Fexofenadine is a nonsedating antihistamine approved for treatment of seasonal allergic rhinitis. OBJECTIVE: This dose-finding study assessed the safety and efficacy of fexofenadine in chronic idiopathic urticaria. METHODS: The 4-week, double-blind, randomized, placebo-controlled study included patients diagnosed with chronic urticaria who had moderate to severe pruritus. Patients received twice daily oral doses of placebo or fexofenadine HCl (20, 60, 120, or 240 mg) at 7 AM and 7 PM. Patients recorded scores for pruritus severity and number of wheals (over the previous 12 hours) in a daily diary. Efficacy variables included mean daily changes from baseline in pruritus severity, number of wheals, and interference with sleep and daily activities due to urticaria. RESULTS: Patients (N = 418) from 37 investigative sites were included. All four fexofenadine HCl doses were statistically superior to placebo (P < or = .0115) for reducing pruritus and number of wheals scores over the 4-week treatment period. There were greater reductions in urticaria symptoms in the 60 mg fexofenadine HCl group than in the 20 mg group, while similar reductions were observed in the 60, 120, and 240 mg dose groups. Additionally, patients receiving fexofenadine experienced significantly less interference with sleep and daily activities than patients receiving placebo (P < or = .0014). Adverse events occurred with similar incidence in all treatment groups, with no dose-related increases in any event. CONCLUSIONS: Fexofenadine HCl significantly reduced pruritus severity, number of wheals, and interference with sleep and normal daily activities in patients with chronic urticaria compared with placebo. Twice-daily doses of 60 mg or greater were most effective. PMID- 10831006 TI - Evaluation of physician and patient compliance with the use of peak flow meters in commercial insurance and Oregon health plan asthmatic populations. AB - BACKGROUND: Home use of peak expiratory flow (PEF) meters is recommended by NIH management guidelines for patients with moderate to severe asthma. OBJECTIVE: To evaluate whether differences in physician PEF meter prescription patterns and patient compliance rates with PEF meter use exist between populations of commercially insured and Oregon Health Plan (OHP) patients. METHODS: A prospective, case-series study was conducted at the Bay Clinic Coos Bay, Oregon. Volunteers responded to a short survey. Patient asthma severity was evaluated by responses to the following questions: (1) frequency of shortness of breath per week, (2) daily and weekly frequency of albuterol rescue inhaler use, and (3) other prescription medications used. Additional questions included: (1) possession of a PEF meter, (2) physician instruction of meter use, (3) patient compliance with instructions, and (4) type of medical insurance. Using NIH asthma stratification guidelines, only respondents with moderate to severe asthma were included in this study. Respondents were age restricted to between 6 and 60 years. Incomplete surveys were not accepted. RESULTS: Ninety-six surveys were received and 54 accepted. Only 24 (44%) of the 54 respondents with moderate to severe asthma had a PEF meter. Nineteen (79%) used it according to physician instructions. Of the 29 patients reporting to have commercial insurance, 18 (62%) had a PEF meter and 15 (83%) used it as prescribed. Of the 21 asthmatic patients with OHP coverage, only 5 (24%) P = .007 had a peak flow meter and 4 (80%) used it as prescribed. CONCLUSIONS: We conclude physician prescription compliance of peak flow meters for asthma monitoring falls short of the NIH guidelines. When a PEF meter is prescribed, patients will likely use it regardless of their medical insurance. Significant differences exist in physician compliance with PEF meter prescription between populations of commercially insured and OHP patients. More study is necessary to further understand this divergence in care delivery. PMID- 10831007 TI - Pharmacokinetics of flunisolide administered via metered dose inhaler with and without a spacer device and following oral administration. AB - BACKGROUND: After inhalation of a glucocorticoid from a meter dose inhaler (MDI), a certain portion of the delivered dose is deposited in the lungs, and the remainder is deposited in the oropharynx. OBJECTIVE: To examine the absolute bioavailability of flunisolide given orally via metered dose inhaler, and metered dose inhaler with a commercially available spacer device as well as to determine the fraction of drug deposited in the lungs following inhalation. METHODS: Twenty four healthy volunteers were enrolled in the study; twenty-two completed the study. The IRB approved the study protocol, and informed consent was obtained. Volunteers received four treatments: treatment A (MDI), 1.0 mg inhaled flunisolide; treatment B (MDI-S), 1.0 mg inhaled flunisolide with a spacer device; treatment C, 1.0 mg of orally administered flunisolide with 240 mL of water; and treatment D, 1.0 mg intravenous flunisolide by IV push in the antecubital vein over 60 seconds. Plasma and urine flunisolide were quantified by HPLC/mass spectrometry/mass spectrometry. RESULTS: Flunisolide is a corticosteroid with low oral bioavailability (6.7%), which was found to be lower than previously reported. Similar AUCs were observed between the MDI and MDI-S groups, but by using mass balance equations, it appears that more flunisolide was delivered to the lungs in the MDI-S group (410 microg versus 280 microg). Oropharyngeal deposition was an important difference between the two inhaler groups. Approximately an 11-fold reduction in the oropharyngeal deposition of flunisolide through use of the spacer device was observed. CONCLUSIONS: Use of a spacer device improved pulmonary delivery of flunisolide by almost 50% and significantly decreased the oropharyngeal exposure to drug. PMID- 10831008 TI - Triamcinolone acetonide aqueous nasal spray versus loratadine in seasonal allergic rhinitis: efficacy and quality of life. AB - BACKGROUND: The new aqueous formulation of triamcinolone acetonide (TAA) was compared with loratadine in patients with seasonal allergic rhinitis. OBJECTIVE: The primary objective of the study was to compare the safety and efficacy of TAA aqueous with loratadine in relieving the symptoms of seasonal allergic rhinitis. METHODS: A total of 351 patients were enrolled into this 4-week, double-blind, double-dummy, randomized, parallel group study. Patients received either TAA aqueous nasal spray (220 microg) or loratadine (10 mg) once daily. Efficacy variables were rhinitis symptom changes from baseline, physician global evaluations, and the patient dropout rate due to insufficient treatment effect. Safety and quality of life also was evaluated. RESULTS: Both TAA aqueous-treated and loratadine-treated patients had improvement in symptoms as early as day 1. Overall, TAA aqueous was significantly (P < .05) more effective than loratadine in reducing total nasal symptoms and individual symptoms of nasal congestion, nasal itch, and sneezing. Ocular symptoms improved from baseline in both groups. There was no statistically significant difference between groups based on physician global evaluation. A similar number of patients in each group discontinued the study due to ineffective treatment. Triamcinolone acetonide aqueous patients maintained a significantly (P < .05) better quality of life in three of the dimensions (activity, nasal symptoms, and practical problems) and for overall quality of life. There were no differences between the two treatment groups in the incidence of adverse events, none of which were clinically significant. CONCLUSIONS: Both TAA aqueous and loratadine were effective and well tolerated in the treatment of patients with seasonal allergic rhinitis. Triamcinolone acetonide aqueous was significantly (P < .05) more effective than loratadine in controlling nasal symptoms of seasonal allergic rhinitis and maintaining a better quality of life for the patients. PMID- 10831009 TI - Circulating ICAM-1 levels in bronchial asthma and the effect of inhaled corticosteroids. AB - BACKGROUND: Even mild asthma has an inflammatory component. Intercellular adhesion molecule-1 (ICAM-1) plays an important role in inflammation. Higher levels of circulating ICAM-1 (cICAM-1) in sera may reflect the upregulation of ICAM-1 expression in allergic inflammation. OBJECTIVE: The aim of this study was to assess cICAM-1 levels in children with atopic bronchial asthma and to determine the effects of inhaled glucocorticoids on cICAM-1 levels. METHODS: The study group consisted of 25 children with moderate atopic bronchial asthma with a mean age of 9.9 +/- 4.2 years, and the control group consisted of 18 healthy children with a mean age of 9.6 +/- 2.6 years. Serum cICAM-1 levels and pulmonary function tests were measured. The patients were treated 2 months with inhaled budesonide 400 to 800 microg (mean 440 microg) per day. After this treatment, cICAM-1 levels and pulmonary function tests were compared with pretreatment levels. The pretreatment cICAM-1 values were compared with healthy control group. RESULTS: The initial cICAM-1 levels of the patient group were significantly higher than the cICAM-1 levels of the control group (P = .001). The post treatment cICAM-1 levels of the patient group were significantly lower than the pretreatment values (P = .007). Pulmonary function test results (FEV1, FEF25-75, and PEFR) rose significantly with the treatment in patients (P < .05). CONCLUSION: This study revealed the presence of inflammation in children with even moderate atopic asthma as reflected with elevated levels of cICAM-1 levels, which decreased following corticosteroid treatment as a result of decreased inflammation. PMID- 10831010 TI - Two-spotted spider mite (Tetranychus urticae): an important allergen in asthmatic non-farmers symtomatic in summer and fall months. AB - BACKGROUND: The Tetranychus urticae (two-spotted spider mite, TSM) is a spider mite commonly found on fruit trees, herbaceous plants, and greenhouse flowers. OBJECTIVE: To evaluate the clinical significance of TSM-derived allergens in non farmers with asthma living around pear orchards, and to assess the allergenic relationship with house dust mite. METHODS: Skin prick tests with TSM were performed in 50 asthmatic non-farmers living around pear orchards. The serum TSM specific IgE was measured in the 16 asthmatics with a positive skin response to TSM. To diagnose TSM-induced asthma, specific bronchial challenges with TSM were performed in the 16 asthmatics. ELISA inhibition tests were performed to assess the allergenic cross-reactivity with house dust mites. RESULTS: Ten of the 16 asthmatics with positive skin responses to TSM showed a significant bronchoconstriction following inhalation of TSM. Eight of the ten TSM-induced asthmatics had high serum TSM-specific IgE and one showed a positive skin response only to TSM. Their asthmatic symptoms were aggravated seasonally, especially in summer and early fall. On ELISA inhibition tests, partial inhibitions with D. pteronyssinus antigens were noted in sera from five asthmatics with positive skin responses to both TSM and D. pteronyssinus, although no inhibition was noted in serum from an asthmatic with a positive skin response only to TSM. CONCLUSION: Tetranychus urticae may be an important allergen in asthmatic non-farmers living around pear orchards and TSM extracts contain species-specific allergens as well as commonly shared allergens with house dust mite. PMID- 10831011 TI - Identification of allergens involved in occupational asthma due to carmine dye. AB - BACKGROUND: Carmine has been implicated as an etiologic agent of occupational asthma, but the allergens involved have not been yet identified. OBJECTIVE: To identify the allergens involved in occupational asthma due to carmine dye. METHODS: An in vitro study based in SDS-PAGE and IgE immunoblotting with carmine and cochineal extracts was performed. Sera from three carmine dye workers diagnosed with occupational asthma induced by carmine dye and from seven nonatopic subjects were used. RESULTS: Three proteins of around 30, 28, and 17 kD in raw cochineal extract and another protein of 50 kD in the boiled one were demonstrated by SDS-PAGE. Two proteins of around 50 and 28 kD were observed in the carmine extract by the same technique. Specific IgE binding bands at 17 kD in cochineal raw extract, at 50 kD in the boiled one, and at 28 kD in carmine extract were demonstrated by IgE immunoblotting. CONCLUSIONS: We have identified three allergens of around 17, 28, and 50 kD implicated in occupational asthma of three carmine workers. PMID- 10831012 TI - Spontaneous expression of mRNA for IL-10, GM-CSF, TGF-beta, TGF-alpha, and IL-6 in peripheral blood mononuclear cells from atopic dermatitis. AB - BACKGROUND: Monocytes and T helper cells play major roles in the immunologic dysfunction of atopic dermatitis (AD). There have been many studies on the cytokine pattern to evaluate abnormalities of immune cells in AD, but the results were conflicting and most of these previous reports were performed with various mitogen-stimulation. OBJECTIVE: The purpose was to investigate de novo cytokine pattern in AD peripheral blood mononuclear cells (PBMC). We focused on the expression of cytokines that have effects on monocytes and T cells. METHODS: We measured mRNA expression of IL-10, GM-CSF, TGF-beta, TNF-alpha, and IL-6 in freshly isolated PBMC with semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR). The intensity of cytokine cDNA were normalized to that of beta-actin product as a standard marker. RESULTS: Interleukin-10 mRNA expression was significantly enhanced in AD compared with control subjects (P < .05). Spontaneous mRNA expression of TGF-beta and TNF-alpha was significantly lower in AD patients (P < .01). The level of GM-CSF mRNA expression was heterogenous and spontaneous mRNA expression was slightly increased in AD although the difference didn't reach the statistical significance. Interleukin-6 mRNA was not detected in most of AD and controls. CONCLUSION: Our data could represent in vivo cytokine expression state associated with monocytes and other immune cells. Increased expression of IL-10 and GM-CSF may be associated with monocyte dysfunction in AD although increase in the expression of GM-CSF mRNA was not statistically significant. Inhibitory effect of increased IL-10 was suggested on decreased expression of TNF-alpha mRNA. The role of TGF-beta in AD remains to be seen. PMID- 10831013 TI - Anaphylaxis to dill. AB - BACKGROUND: Allergic symptoms caused by spices and herbs are infrequent and usually mild, although occasionally, severe allergic reactions do occur. Symptoms of pruritus, rhinitis, cough, and edema have been reported to spices including curry, paprika, pepper, and mustard. To our knowledge, this is the first case of confirmed dill allergy, and the patient had severe allergic symptoms. OBJECTIVE: It is important to alert physicians to the possibility of allergic reactions caused by dill. METHODS AND RESULTS: The patient, who has a history of allergic rhinitis, developed symptoms of oral pruritus, tongue and throat swelling, urticaria, and immediate vomiting and diarrhea following ingestion of foods cooked with dill and subsequently with inhalation of foods prepared with dill. Skin testing with fresh dill preparation was positive. CONCLUSION: These findings confirm that dill can cause IgE-mediated reactions. PMID- 10831014 TI - Drug transporters in psychopharmacology--are they important? PMID- 10831015 TI - Longitudinal comparative study of risperidone and conventional neuroleptics for treating patients with schizophrenia. The Quebec Schizophrenia Study Group. AB - This study compared the long-term (12 months) effectiveness of risperidone (RP) with that of conventional neuroleptics (CNs) in a population with chronic schizophrenia who had shown suboptimal response to CNs. A randomized, open, parallel, multicenter design was used. One hundred eighty-four subjects meeting DSM-IV criteria for schizophrenia were randomly assigned to receive either RP or a CN, and 165 of them completed the follow-up. Outcome measures were taken at 3, 6, and 12 months and included the Positive and Negative Syndrome Scale (PANSS) and the Extrapyramidal Symptom Rating Scale. Within this 12-month follow-up, RP was found to be superior to CNs in terms of both the average change in score from baseline on the PANSS (p = 0.006) and the proportion of good responders (as defined by a 20% decrease in total PANSS scores;p = 0.03). For positive symptoms, the effectiveness of the RP treatment tended to increase over time. At 12 months, the percentage of good responders in the RP group was twice as large as that in the CN group (30% vs. 15%;p = 0.03). The superiority of RP over CNs was constant over the three dose categories. In both the RP and the CN groups, the maximum decrease in psychopathology was achieved with the lowest dose range. A worsening of akathisia was less frequent in subjects receiving RP than in those receiving CNs (p = 0.02). In conclusion, this study showed that, compared with CNs, RP is beneficial in the treatment of patients with chronic schizophrenia and that some of these benefits may appear only after longer-term treatment. PMID- 10831016 TI - Risperidone for the treatment of cocaine dependence: randomized, double-blind trial. AB - A partial blockade of the multiple actions of cocaine is one strategy by which cocaine dependence may be treated. Risperidone, a 5-hydroxytryptamine and dopamine D2 antagonist, is an atypical antipsychotic and was a candidate medication for the treatment of cocaine dependence. One hundred ninety-three cocaine-dependent subjects were enrolled in a 12-week, randomized, double-blind, placebo-controlled trial. Subjects initially received either placebo or 4 or 8 mg of risperidone, with a subsequent change to active doses of 2 mg and 4 mg. Subjects attended the clinic twice each week, provided urine samples, obtained medication, and underwent one behavioral therapy session per week. The study was terminated at the interim analysis. Retention was worse for the 4- and 8-mg active medication groups. Side effects were primarily associated with the 8-mg dose, although neither 2 mg nor 4 mg was well accepted by subjects. There was no reduction in cocaine use associated with risperidone. The results suggest that although antagonists might be a useful treatment approach, such as in the treatment of opiate dependence, risperidone is unlikely to find broad acceptance with the treatment-seeking population. PMID- 10831017 TI - Anticholinergic differences among patients receiving standard clinical doses of olanzapine or clozapine. AB - This study evaluated anticholinergic effects among patients with schizophrenia, schizoaffective disorder, or bipolar I disorder who were receiving either olanzapine (N = 12) or clozapine (N = 12) at standard clinical doses in a naturalistic setting. Serum anticholinergic levels were determined in adult male and female subjects using a radioreceptor binding assay. The Udvalg for Kliniske Undersogelser Scale was used to evaluate anticholinergic side effects clinically, and the Mini-Mental State Examination provided a global cognitive measure. Patients had achieved target doses that were stable at the time at which blood samples were obtained, and no other concomitant medicine with known anticholinergic potential was allowed. Patients receiving olanzapine (average dose, 15 mg/day) had serum anticholinergic levels of 0.96 (+/-0.55) pmol/ atropine equivalents compared with levels of 5.47 (+/-3.33) pmol/atropine equivalents for those receiving clozapine (average dose, 444 mg/day) (p < 0.001). Rates of increased and decreased salivation were significantly more common among the clozapine- and olanzapine-treated patients, respectively, whereas constipation, urinary disturbances, and tachycardia/palpitations were significantly more common among clozapine-treated patients. Neither group showed any global cognitive deficits. Olanzapine-treated patients had serum anticholinergic levels that were less than one fifth those of the clozapine treated patients. Furthermore, clinical evaluations confirmed that clozapine treated patients experienced more frequent and severe anticholinergic side effects (except dry mouth). However, none of the patients in either group expressed any desire to discontinue these medications as a result of the anticholinergic side effects. PMID- 10831018 TI - Studies on the mechanism of a fatal clarithromycin-pimozide interaction in a patient with Tourette syndrome. AB - The authors report in detail the case of a 27-year-old man who experienced sudden cardiac death 2 days after coprescription of the neuroleptic pimozide and the macrolide antibiotic clarithromycin after the documentation of a prolonged QT interval. To determine the prevalence of this interaction, the authors referred to the Spontaneous Reporting System of the Food and Drug Administration and identified one similar case in which clarithromycin was coprescribed with pimozide and sudden cardiac death occurred shortly thereafter. In addition, the search identified 39 cases of cardiac arrhythmia associated with pimozide, 11 with pimozide alone, and 6 with clarithromycin alone, 1 of which had a positive rechallenge. The mechanism of the interaction between clarithromycin and pimozide seems to involve the inhibition of the hepatic metabolism of pimozide by the macrolide. The authors demonstrated that clarithromycin is able to inhibit the metabolism of pimozide in human liver microsomal preparations (K(i) = 7.65 +/- 1.18 microM) and that pimozide, but not clarithromycin or its primary metabolite, is able to prolong the electrocardiac QT interval in a dose-dependent manner in the isolated perfused rabbit heart. The increase was 9.6 +/- 1.1% in male hearts (N = 5) and 13.4 +/- 1.2% in female hearts (N = 4) (p < 0.05). PMID- 10831019 TI - Sudden death in patients receiving clozapine treatment: a preliminary investigation. AB - The risk of sudden death during clozapine treatment is controversial. The authors present a review of sudden deaths that occurred at Sha'ar Menashe Mental Health Center between January 1991 and August 1997. The number of cases of deceased inpatients was retrieved from the hospital's computerized database and divided into three groups: sudden death, suicide, and disease-related death. Copies of mandatory reports of sudden death filed with the Ministry of Health were obtained, and the corresponding patient records were reviewed. The rates of sudden death, suicide, and disease-related deaths were calculated for clozapine treated patients (CTPs) during and after treatment and for patients treated with other psychiatric agents (non-CTPs). Among 561 CTPs, there were 4 sudden deaths during treatment, 2 sudden deaths after treatment, 2 suicides during treatment, and 2 disease-related deaths during treatment. Among 4918 non-CTPs, there were 14 sudden deaths, 5 suicides, and 86 disease-related deaths, all of which occurred during treatment with other psychiatric agents. CTPs who experienced sudden death were 10.37 years younger and healthier than non-CTPs who experienced sudden death. The sudden death rate was 3.8 times higher for CTPs than for non-CTPs, whereas the rate of disease-related death was 5 times higher for non-CTPs than for CTPs. Contrary to expectations, the rate of suicide among patients currently receiving clozapine in this sample was 3.6 times higher than among non-CTPs. Because CTPs who experienced sudden death were also younger and healthier, it seems that treatment with clozapine may present a greater risk for sudden death than treatment with other psychiatric medications. The limited number of sudden death cases and deaths from other causes should be noted so that these findings are considered with caution. PMID- 10831020 TI - Comparison of the effects of zaleplon, zolpidem, and triazolam on memory, learning, and psychomotor performance. AB - Twenty-four healthy male and female subjects, who participated in this randomized, double-blind, crossover study, received single nighttime doses of zaleplon 10 mg (therapeutic dose), zaleplon 20 mg, zolpidem 10 mg (therapeutic dose), zolpidem 20 mg, triazolam 0.25 mg (positive control), and placebo. Subjective behavioral ratings and psychomotor tests were completed before and 1.25 and 8.25 hours after administration of the study drug. The Immediate and Delayed Word Recall tests and the Digit Span Test were used to assess memory. The Digit-Symbol Substitution Test, Paired Associates Learning Test, and Divided Attention Test were used to assess other cognitive skills. Zaleplon 10 mg did not produce any significant changes in memory or learning compared with placebo. All other active treatments, including zolpidem 10 mg, caused psychomotor impairment at the 1.25-hour test battery. Zolpidem 20 mg (twice the therapeutic dose) produced more psychomotor impairment at the 1.25-hour assessment than did any of the other active treatments, including zaleplon 20 mg. At the 8.25-hour time point, test scores for subjects who received zaleplon 10 mg and 20 mg did not differ from the test scores for those who received placebo. However, cognitive impairment persisted up to the 8.25-hour observation for subjects who were administered triazolam 0.25 mg and zolpidem 20 mg. Adverse events associated with the use of zaleplon were transient and mild-to-moderate in severity. Overall, this study shows that zaleplon is a safe hypnotic that does not affect memory, learning, or psychomotor skills associated with vigilance. PMID- 10831021 TI - Differences in pharmacodynamics but not pharmacokinetics between subjects with panic disorder and healthy subjects after treatment with a single dose of alprazolam. AB - The pharmacokinetics and pharmacodynamics of the benzodiazepine alprazolam (1 mg, administered orally) were compared between eight patients with panic disorder and eight age- and sex-matched healthy volunteers. Subjects received orally administered placebo and alprazolam in a randomized, double-blind, single-dose crossover study. The elimination half-life, time of maximum plasma concentration, maximum concentration, volume of distribution, and clearance of alprazolam were similar for both groups. For each cohort, alprazolam treatment (vs. placebo) produced significant changes in typical benzodiazepine agonist effects, such as increased sedation and impaired cognitive performance on the digit-symbol substitution test. For the panic disorder group only, there was a significant increase in the subjective rating of"contented" and a reduction in the rating of "easily irritated." For the healthy volunteer group, alprazolam produced increases in ratings of "fatigued" and "slowed thinking," but also increases in ratings of "relaxed." In each group, alprazolam significantly increased the electroencephalographic (EEG) measure of relative beta amplitude (range, 13-30 Hz) compared with placebo. Concentration-EEG response curves fit a sigmoid E(max) model, and there was greater sensitivity to EEG effects, as measured by a 28% reduction in the EC50 value, in the panic disorder group compared with healthy control subjects. After alprazolam treatment, there was increased sensitivity to EEG and mood effects and fewer aversive effects in the panic disorder group compared with healthy subjects. There were no differences in the pharmacodynamic measures of sedation and cognition or differences in pharmacokinetics between the two groups. PMID- 10831022 TI - Single-dose pharmacokinetics of methylphenidate in CYP2D6 extensive and poor metabolizers. AB - Six adults phenotyped as either extensive (N = 4) or poor (N = 2) metabolizers for cytochrome P450 (CYP) 2D6 were given a 10-mg oral dose of methylphenidate (MPH) on two separate occasions with and without quinidine, a potent CYP2D6 inhibitor. Quinidine had no significant effect on the pharmacokinetics of either MPH or ritalinic acid, its major metabolite, in either group of CYP2D6 metabolizers. These data suggest a lack of involvement of CYP2D6 in the metabolism of MPH. Drugs that are inhibitors of CYP2D6 when taken concurrently with MPH should not affect its plasma concentration. PMID- 10831023 TI - A 12-week, open trial of donepezil hydrochloride in patients with multiple sclerosis and associated cognitive impairments. AB - Cognitive dysfunction occurs in up to 65% of patients with multiple sclerosis (MS), but there is no effective treatment for the symptoms. The authors conducted a 12-week, open-pilot study to assess the efficacy and tolerability of donepezil HCl administered in patients with MS and cognitive impairment. Seventeen patients at a long-term care facility with Mini-Mental State Examination scores of < or = 25 received 5 mg of donepezil HCl for a 4-week period, followed by 8 weeks of 10 mg of donepezil HCl. Cognitive, neurologic, functional, and behavioral assessments were conducted at baseline and at 4 and 12 weeks. Statistically significant improvement was observed in several cognitive domains including attention, memory, and executive functioning, as well as different aspects of behavior. These data suggest that donepezil HCl merits further study as a potentially viable treatment option for patients with cognitive impairment associated with MS. PMID- 10831024 TI - Randomized, placebo-controlled pilot study of divalproex sodium in the treatment of acute exacerbations of chronic schizophrenia. AB - Experimental and clinical data suggest that GABA-ergic drugs such as valproate may have a potential role in the treatment of schizophrenia. The authors designed a 21-day prospective, double-blind, randomized, placebo-controlled pilot study of divalproex sodium as add-on treatment to haloperidol in 12 hospitalized patients with acute exacerbations of chronic schizophrenia. All patients received haloperidol 10 mg/day for 3 days and 15 mg/day for the remaining 18 days. In addition, five patients were randomly assigned to receive divalproex augmentation and seven to receive placebo. The divalproex dose was adjusted to a target serum concentration of 75 microg/mL for 2 weeks; placebo replaced divalproex during the third and last weeks to determine any carryover effect. Psychiatric rating scales were administered at baseline and on days 7, 14, and 21. Although the placebo group improved with haloperidol treatment, the divalproex group demonstrated greater improvement. On day 21, the divalproex group had greater improvement from baseline on the Clinical Global Impression Scale (p < or = 0.04), Brief Psychiatric Rating Scale (p < or = 0.13), and Schedule for Assessment of Negative Symptoms scores (p < or = 0.007). After divalproex withdrawal on day 15, a carryover effect was observed during week 3. The authors concluded that the addition of divalproex sodium to standard antipsychotic drugs may prove effective in relieving the symptoms of acute schizophrenia. Future studies may benefit from the design of this pilot study. However, it is premature to apply this augmentation strategy in the clinical setting just yet because of the small sample size and the likely heterogeneity of the disorder. PMID- 10831025 TI - Placebo-controlled study of fluvoxamine in the treatment of patients with compulsive buying. AB - Compulsive buying is a syndrome characterized by the impulsive and/or compulsive buying of unneeded objects that results in personal distress, impairment in vocational or social functioning, and/or financial problems. Results from a two site, double-blind, placebo-controlled 13-week trial of fluvoxamine are presented. Subjects had problematic buying behavior that they could not control for the previous 6 months or longer and met DSM-IV criteria for impulse control disorder-not otherwise specified (ICD-NOS) and the University of Cincinnati criteria for compulsive buying. Assessments included clinician-rated scales-the Yale-Brown Obsessive Compulsive Scale modified for compulsive buying, the Clinical Global Impression Scale, the Global Assessment of Functioning, and the Hamilton Rating Scale for Depression-and patient self-reports using daily diaries, which measured episodes of compulsive buying. Forty-two subjects gave informed consent, with 37 subjects providing evaluable information and 23 completing the study. Current or past psychiatric comorbidity was present in 74% of subjects. Intent-to-treat and completer analyses failed to show a significant difference between treatments on any measures of outcome. A high placebo-response rate, possibly from the behavioral benefits of maintaining a daily diary, prevents any definitive statement on the efficacy of fluvoxamine in treating compulsive buying. PMID- 10831026 TI - Fluvoxamine reduces physiologic reactivity to trauma scripts in posttraumatic stress disorder. AB - This study assessed the effect of open-label fluvoxamine treatment for posttraumatic stress disorder (PTSD), depressive symptoms, and physiologic arousal to trauma cues. Baseline psychometric ratings and physiologic assessments of heart rate and blood pressure responses to individualized, taped trauma scripts were determined for 16 patients with PTSD and 16 mentally healthy age- and gender-matched control subjects exposed to at least 1 serious trauma. Patients with PTSD had greater autonomic reactivity than control subjects at baseline, with physiologic measures correlating with the severity of the PTSD for the combined groups. Discriminant analyses indicated that systolic blood pressure best classified patients with PTSD (75% sensitivity) and control subjects (100% specificity), with a stepwise discriminant analysis showing that combined physiologic variables correctly classified 75% of patients with PTSD and 100% of control subjects. After 10 weeks of fluvoxamine treatment (100-300 mg/day), patients' PTSD, depression, and physiologic reactivity improved significantly. Medicated patients with PTSD could not be distinguished statistically from untreated control subjects in any physiologic measure. This dampening of autonomic reactivity after drug treatment corroborates subjective measures of improvement, validating the reported efficacy of fluvoxamine in this open trial. PMID- 10831027 TI - Does the addition of pindolol accelerate the response to electroconvulsive therapy in patients with major depression? A double-blind, placebo-controlled pilot study. AB - There is evidence that addition of pindolol, a beta-adrenergic/5 hydroxytryptamine-1A antagonist, can accelerate the onset of action of antidepressant medications. The purpose of this study was to determine whether pindolol administration can induce a rapid improvement in depressive symptoms in patients receiving electroconvulsive therapy (ECT) within six ECT treatments. A total of 20 patients with DSM-IV-diagnosed major depression who were undergoing a course of ECT as the clinically indicated treatment were recruited. They were neuroleptic, lithium, and antidepressant free for at least 1 week before the study. Of the 20 patients, 9 patients had been randomly assigned to receive pindolol 2.5 mg three times daily, and 11 patients received identical placebo three times daily for the duration of the first 6 ECT treatments. One of 9 patients in the pindolol group and 4 of 11 patients in the placebo group dropped out of the study. Using an outcome measure of a score < or =12 on the 29-item Hamilton Rating Scale for Depression (HAM-D), the authors found that four (50%) of eight patients responded to the combination treatment of ECT and pindolol within six ECT treatments. In contrast, none (0%) of seven patients who received placebo responded to ECT treatment. Furthermore, both mean 29-item HAM-D and Clinical Global Impression Scale scores after the sixth ECT treatment were significantly lower in patients treated with pindolol compared with those treated with placebo. However, the number of total ECT treatments within a course or the overall efficacy of ECT treatment was not altered by the addition of pindolol. The results of this study suggest that within six ECT treatments, pindolol administration hastens antidepressant effects of ECT in some depressed patients. PMID- 10831028 TI - Anorgasmia in a patient with bipolar disorder type 1 treated with gabapentin. AB - Gabapentin is a relatively new anticonvulsant indicated for adjunctive therapy in the treatment of partial seizures, with and without secondary generalization, in adults with epilepsy. Overall, it has a minimal side effect profile compared with other anticonvulsant agents. Postmarketing surveillance is needed to further delineate the spectrum of adverse events that may be experienced by patients treated with this medication. This is a case report of a 25-year-old man with a 10-year history of mood swings that progressively worsened and resulted in a suicide attempt 8 months before his first appointment. A diagnosis of bipolar disorder was established, and a clinical interview ruled out other mental disorders. The patient was administered gabapentin 300 mg/day, and the dose was titrated upward to 900 mg/day. A follow-up appointment revealed improved control of his bipolar symptoms. However, the patient reported that he could not have an orgasm during sexual intercourse. The medication was changed to valproic acid 250 mg three times daily. His bipolar symptoms remained under control and the anorgasmia resolved. This was maintained at the next follow-up appointment. The side effect profile and therapeutic monitoring requirements of gabapentin are favorable when compared with those of other anticonvulsant agents. However, because this agent is relatively new, especially for use in the treatment of bipolar disorder, a more thorough development of its side effect profile is needed. Observing, recording, and reporting atypical adverse events and side effects are critical to postmarketing surveillance and enhance the clinician's ability to make rational therapeutic decisions. PMID- 10831029 TI - Adverse effects of high-dose olanzapine in treatment-refractory schizophrenia. PMID- 10831030 TI - Case of pick's central lobar atrophy with apparent stabilization of cognitive decline after treatment with risperidone. PMID- 10831031 TI - Effect of adjunctive treatment with serotonin-1A agonist tandospirone on memory functions in schizophrenia. PMID- 10831032 TI - Nonconvulsive status epilepticus complicating neuroleptic malignant syndrome improved by intravenous diazepam. PMID- 10831033 TI - Failure to consider drug-drug interactions as a likely cause of behavioral deterioration in a patient with bipolar disorder. PMID- 10831034 TI - Potential drug interaction between lithium and valsartan. PMID- 10831035 TI - Olanzapine treatment of methamphetamine psychosis. PMID- 10831036 TI - Tolerability and efficacy of high-dose moclobemide alone and in combination with lithium and trazodone. PMID- 10831037 TI - Treating Charles Bonnet syndrome: understanding inconsistency. PMID- 10831038 TI - Functional retinoid receptors in budding ascidians. AB - A homolog of retinoid X receptors (RXR), named PmRXR, was cloned from the budding ascidian, Polyandrocarpa misakiensis. Gel-shift assays revealed that PmRXR and a previously identified P. misakiensis retinoic acid receptor (PmRAR) formed a complex to bind vertebrate-type retinoic acid response element (RARE). Transfection assays were carried out using a reporter gene containing a RARE upstream of lacZ. Two chimeric effector genes were constructed by placing PmRXR and PmRAR cDNA fragments (containing the DNA-binding, ligand-binding and ligand dependent transactivation domains) downstream of the human RXR alpha and RAR alpha cDNA (covering the N-terminal coding region), respectively. Each chimeric cDNA was ligated to a notochord-specific enhancer. In case the embryos were transfected with all three transgenes and treated with retinoic acid (RA), the reporter gene was activated in the notochord cells. The result suggests that the PmRXR/PmRAR complex functions as an RA-dependent transcriptional activator. The PmRXR mRNA was detected in a mesenchymal cell type, called glomerulocyte, in the developing Polyandrocarpa bud. As this cell type has been shown to express PmRAR mRNA, it seems possible that the PmRXR/PmRAR complex mediates RA signaling in this cell type to induce the expression of genes involved in the morphogenesis of the developing bud. PMID- 10831039 TI - Primary mesenchyme cell-ring pattern formation in 2D-embryos of the sea urchin. AB - Primary mesenchyme cell (PMC) migration during PMC-ring pattern formation was analyzed using computer-assisted time-lapse video microscopy in spread embryos (2D-embryo) of the sea urchin, Mespilia globulus, and a computer simulation. The PMC formed a near normal ring pattern in the 2D-embryos, which were shown to be an excellent model for the examination of cell behavior in vivo by time-lapse computer analysis. The average migration distance of the ventro-lateral PMC aggregate-forming cells (AFC) and that of the dorso-ventral PMC cable-forming cells (CFC) showed no significant difference. All PMC took a rather straightforward migration path to their destinations with little lag time after ingression. This in vivo cell behavior fitted well to a computer simulation with a non-diffusable chemotaxis factor in the cyber-cell migration field. This simulation suggests that PMC recognize their destination from a very early moment of cell migration from the vegetal plate, and implicates that a chemoattractive region is necessary for making the PMC migration pattern. The left- and right lateral AFC and dorso and ventral CFC were each derived from an unequally divided one-quarter segment of the vegetal plate. This suggests that AFC and CFC have a distinctive ancestor in the vegetal plate, and the PMC are a heterogeneous population at least in terms of their destination in the PMC-ring pattern. PMID- 10831040 TI - Appearance of hair follicle-inducible mesenchymal cells in the rat embryo. AB - Rat vibrissa follicle morphogenesis starts around 13 days of gestation. By day 14 mesenchymal cells have already aggregated as 'condensations' beneath the initial hair bud. Some of the mesenchymal cells will form a dermal papilla, having profound effects on hair follicle formation. The appearance of follicle-inducing mesenchymal cells in the process of vibrissa follicle development was examined. Mesenchymal cells were isolated from the developing site of vibrissa follicles at 13 days or at later stages and amplified in mass culture, harvested and transplanted in association with the epithelium. It was demonstrated that 13-day mesenchymal cells did not induce any hair bulbs but those from 14 days or later stages could induce hair-producing new bulbs or new follicles depending on the association with the follicle epithelium or with the glabrous sole epidermis of the adult rats, respectively. Further, clones having hair bulb-inducing ability were obtained from 14- and 15-day mass-cultured mesenchymal cells. Based on these and other results, it was concluded that mesenchymal cells having follicle inducing ability are present at least by 14 days in the future whisker pad region. This suggests that the differentiation of the dermal papilla cells must start before the initial hair bud stage. PMID- 10831041 TI - On the mechanism of ooplasmic segregation in single-cell zebrafish embryos. AB - It has been previously shown that localized elevations of free cytosolic calcium are associated with a morphological contraction in the forming blastodisc and animal hemisphere cortex during ooplasmic segregation in zebrafish zygotes. It was subsequently proposed, in a hypothetical model, that these calcium transients might be linked to the contraction of a cortically located actin microfilament network as a potential driving force for segregation. Here, by labeling single cell embryos during the major phase of segregation with rhodamine-phalloidin, direct evidence is presented to indicate that the surface contraction was generated by an actin-based cortical network. Furthermore, while zygotes incubated with colchicine underwent normal ooplasmic segregation, those incubated with cytochalasin B did not generate a constriction band or segregate to form a blastodisc. During segregation at the single-cell stage, ooplasm simultaneously moved in two directions: toward the blastodisc within the so-called axial streamers, and toward the vegetal pole in the peripheral ooplasm. The velocities of both axial and peripheral streaming movements are reported. By injection of a fluorescein isothiocyanate (FITC)-labeled 2000 kDa dextran into the peripheral ooplasm it was demonstrated that a portion of it feeds into the bases of the extending streamers, which helps to explain the lack of accumulation of ooplasm at the vegetal pole. These new data were incorporated into the original model to link the bipolar ooplasmic movements with the calcium-modulated, actin-mediated contraction of the animal hemisphere cortex as a means of establishing and driving ooplasmic segregation in zebrafish. PMID- 10831042 TI - Establishment of pigment cell lineage in embryos of the sea urchin, Hemicentrotus pulcherrimus. AB - In an attempt to estimate the number of pigment precursor cells in sea urchin embryos, DNA synthesis and cell divisions were blocked with aphidicolin from various stages of development. Interestingly, pigment cells differentiated on a normal time schedule, even if the embryos were treated from late cleavage stages on. In most of the embryos treated from 10 h on, 10-15 pigment cells differentiated. Thereafter, the number of pigment cells in the aphidicolin treated embryos further increased, as the initiation of the treatment was delayed. On the other hand, total cell volumes in the pigment lineage, calculated from the averaged number and diameter of differentiated pigment cells, were almost the same irrespective of the time of the initiation of aphidicolin treatment. This indicated that the increase in the number was caused by divisions of the pre-existing cells in the pigment lineage. Thus, the founder cells that exclusively produce pigment cells could be identified. They are nine times cleaved blastomeres and specified by 10 h post-fertilization. The obtained results also clarified the division schedule in the pigment lineage; the founder cells divide once (10th) until hatching, and divide once more (11th) by the end of gastrulation. PMID- 10831044 TI - A starfish homolog of mouse T-brain-1 is expressed in the archenteron of Asterina pectinifera embryos: possible involvement of two T-box genes in starfish gastrulation. AB - A cDNA clone for a starfish T-box gene (Ap-Tbr) was isolated and characterized. Molecular phylogenetic analysis showed that the Ap-Tbr gene was a member of the T brain subfamily, which includes mouse T-brain-1 and Xenopus Eomesodermin. Ap-Tbr was expressed as early as in late blastulae, and the transcript was evident in a disc-like region at the vegetal end or the vegetal plate. In early gastrulae, the gene was expressed in the cells of the invaginated archenteron, from which the majority of mesodermal cells as well as some endodermal cells are derived. The Ap Tbr expression disappeared by the end of gastrulation, and was not detected in early bipinnaria larvae. This expression pattern of Ap-Tbr suggests its role in an early step of archenteron invagination, associated with mesoderm and endoderm formation. Furthermore, double staining of late blastulae and early gastrulae with a probe specific for Ap-Tbr and one for ApBra (the starfish Brachyury gene) demonstrated that the regions of Ap-Tbr and ApBra expression at the vegetal/posterior end of the embryo did not overlap, the region with Ap-Tbr expression being encircled by a ring-shaped region of ApBra expression. A gap without expression of the two genes was located between them, and the gap was seen around the blastoporal lip in the early gastrula. These observations suggest implication of the two T-box genes, with different roles, in starfish gastrulation. PMID- 10831043 TI - Delocalization of polar plasm components caused by grandchildless mutations, gs(1)N26 and gs(1)N441, in Drosophila melanogaster. AB - Two maternal-effect grandchildless (gs) mutations of Drosophila melanogaster, gs(1)N26 and gs(1)N441, cause delay in nuclear arrival at the polar plasm. In mutant embryos, polar plasm loses its ability to induce pole cells during retarded nuclear migration to the posterior pole of embryos. In the present study, it was shown that in N26 and N441 embryos, mitochondrial large rRNA (mtlrRNA), an essential factor for pole cell formation, is delocalized during the delay in nuclear arrival. This suggests that the loss of mtlrRNA causes failure of the mutants to form pole cells. Furthermore, it was shown that all of the other polar plasm components examined, namely Vasa protein, Germ cell-less protein, nanos mRNA and Polar granule component RNA start to be delocalized during the delay in nuclear arrival. This suggests that polar plasm integrity is not maintained in mutant embryos. It was finally shown that Vas is also delocalized in embryos that are inhibited to form pole cells by reducing the amount of mtlrRNA. This indicates that the segregation of polar plasm into pole cells is required to maintain polar plasm integrity. The mechanism regulating polar plasm integrity in embryos is discussed. PMID- 10831045 TI - Mutations with sensory ray defect unmask cuticular glycoprotein antigens in Caenorhabditis elegans male tail. AB - Caenorhabditis elegans male tail has nine pairs of bilaterally symmetric ray processes extended into a cuticular fan. The formation of these structures involves both cell lineage differentiation and cellular morphogenesis. Nine mutations were examined, all of which presented an amorphous ray phenotype. Glycoconjugates carrying an N-acetylglucosamine (GlcNAc) epitope were detected at a high level in their male bursa. It was shown that these antigens are not responsible for the morphological defects. It was further demonstrated that these ram and mab gene products represent critical components for male tail cuticle organization. Mutations of them abolish the integrity of the male bursal cuticle and unmask the underlying GlcNAc epitope. PMID- 10831046 TI - Lipid dynamics in the embryos of Patiriella species (Asteroidea) with divergent modes of development. AB - Evolution of lecithotrophic development in sea stars involved a modification in maternal provisioning from the production of yolk-dominated to lipid-dominated eggs. The dynamics of lipid reserves in the embryos of four Patiriella species differing in their lipid provisions were examined. Patiriella regularis had small yolk protein-dominated eggs (150 microm in diameter) and an ancestral mode of development through planktotrophic larvae. Patiriella calcar, Patiriella exigua and Patiriella pseudoexigua had large eggs (390-440 microm in diameter) and lecithotrophic planktonic, benthic and intragonadal larvae, respectively. Patiriella exigua deposited negatively buoyant eggs containing substantial yolk protein and lipid reserves onto the substratum. In contrast, the planktonic eggs of P. calcar and the intragonadal eggs of P. pseudoexigua were dominated by lipid and were neutrally and positively buoyant, respectively. By the blastula stage there was little trace of lipid in P. regularis embryos. Blastulae of the lecithotrophic developers, by contrast, had conspicuous lipid droplets distributed through their cells. In parallel with the change from cuboidal to columnar epithelium during the blastula to gastrula transition, lipid reserves became redistributed into the basal cytoplasm. The extent of lipid transport reflected the amount of lipid reserves. In P. pseudoexigua embryos with the greatest lipid load, basal shunting was followed by secretion of lipid into the blastocoele where it was stored for the perimetamorphic period. Evolution of lecithotrophy in Patiriella appears to reflect selection to provide metamorphic stages with nutrients normally accrued by feeding larvae with the consequence that early development is burdened by voluminous, potentially inert nutritive stores. Lipid redistribution coincident with a major developmental stage transition may be required to facilitate unimpeded morphogenesis. This phenomenon may be characteristic of lecithotrophic development in echinoderms and appears pre-adaptive for extrusion of lipid in species like P. pseudoexigua with particularly extensive lipid reserves. PMID- 10831047 TI - Cercarial productivity of redial generations in single-miracidium infections of Lymnaea truncatula with Paramphistomum daubneyi or Fasciola hepatica. AB - Single-miracidium infections of Lymnaea truncatula with Paramphistomum daubneyi or with Fasciola hepatica were carried out under laboratory conditions to count free rediae, their germinal embryos, and to determine the cercarial productivity of each redial generation. In snails infected by P. daubneyi, the cercariae were produced by the first (8.7 cercariae per redia) and second (8.9 per redia) generations. At day 63 post-exposure, they corresponded, respectively, to 53.9% and 46.1% of cercariae produced by all rediae. In snails infected by F. hepatica, the majority of cercariae were produced by the R2a group (18.2 cercariae per redia) and corresponded to 66.0% of cercariae produced all rediae. The cercariae produced by the other redial groups were more limited in number: 17.5 per redia in the R1b group (28.7%) and 2.0 per redia in the R2b/R3a group (5.3%). Cercarial productivity of P. daubneyi until day 63 post-exposure was more limited in number than that of F. hepatica: a total of 145 cercariae per snail versus 427 per snail. PMID- 10831048 TI - Seasonal and site specific variation in the component community structure of intestinal helminths in Apodemus sylvaticus from three contrasting habitats in south-east England. AB - Seasonal fluctuations in the prevalence and abundance of infection with intestinal helminths were studied in Apodemus sylvaticus (wood mouse, n = 399), from three contrasting habitats in southern England, to test the hypothesis that both intrinsic (host sex, age) and extrinsic (season, site) factors influence parasite species richness and abundance. Five species of helminths were recovered but only one of these (Capillaria murissylvatici) was site-specific (Dungeness). Total species richness was therefore 5 at Dungeness and 4 at the other two sites. Mean species richness was 1.4, but in adult mice there was a pronounced difference between the sites, and an independent highly significant effect of season. Syphacia stroma and Corrigia vitta both showed marked differences between sites in respect of prevalence and abundance of infection. Capillaria murissylvatici was encountered at Dungeness mostly in the spring whereas seasonal changes in abundance of S. stroma were consistent across all three sites. Seasonal fluctuations in the abundance of Catenotaenia pusilla were compounded by differences between sites. Host sex was not a significant factor in any species, although a posteriori analysis of S. stroma worm burdens for the Isle of Wight site revealed a moderate local sex effect. Overall the principal determinants of variation in helminth burdens were the extrinsic factors, site and season. PMID- 10831049 TI - Temporal variations in the infection of a population of Cerithidea cingulata by larval trematodes in Kuwait Bay. AB - The prosobranch gastropod Cerithidea cingulata (Gastropoda: Potamididae) in Kuwait Bay was examined for larval trematode infections over a 17-month period. A total of 2537 snails were examined and 1265 (49.9%) found to be infected with one or more species of trematodes. The component community in the snail comprised 12 species representing the families Cyathocotylidae (2), Echinostomatidae (2), Haplosplanchnidae (1), Heterophyidae (2), Microphallidae (1), Philophthalmidae (2), Plagiorchiidae (1) and Schistosomatidae (1). Cyathocotylid II (41.6%) was by far the most prevalent species followed by the microphallid (3.9%), the two species comprised 90% of the total trematode fauna. The prevalence of infection increased with shell size and was significantly higher in male (47%) than female (33%) snails. Multiple infections were observed in only 15 (1.2%) of the infected snails; cyathocotylid I and cyathocotylid II combination occurred 14 times and heterophyid I and the microphallid occurred once. Trematode species were more diverse and prevalent in winter, and cercarial shedding peaked in summer. Behaviour of the definitive host and snail population dynamics were probably the major contributors to the detected temporal pattern in the infections. PMID- 10831050 TI - Opisthorchis viverrini: ultrastructure and cytochemistry of the glycocalyx of the tegument. AB - The ultrastructure and cytochemistry of the glycocalyx of the tegument of Opisthorchis viverrini during maturation from newly excysted juvenile to adult stages were investigated using colloidal iron, ruthenium red and lectin stainings. The results showed that the glycocalyx was intensely stained by the first two dyes, thus indicating the presence of relatively high amounts of negative charges. However, the thickness and intensity of the staining decreased during the fluke's maturation. Binding studies using lectin probes on the surface of adult parasites showed that binding sites for Canavalia ensiformis (Con A), Triticum vulgaris (WGA) and Ricinus communis I (RCA I) were present in relative large amounts on the glycocalyx of the adult tegument, whereas those for Dolichos biflorus (DBA) were relatively fewer in number, and those for Ulex europaeus I (UEA I) were absent. The binding patterns of Con A, WGA, RCA I and DBA were generally similar, and the reaction product was uniformly distributed over the dorsal and ventral surfaces of the parasite's body. These bindings, therefore, indicate the presence of D-mannose/D-glucose, N-acetyl-D-glucosamine/sialic acid, D-galactose and N-acetyl-D-galactosamine residues on the glycocalyx of the adult tegument. PMID- 10831051 TI - Intestinal helminths of spiny mice (Acomys cahirinus dimidiatus) from St Katherine's Protectorate in the Sinai, Egypt. AB - Spiny mice, Acomys cahirinus dimidiatus, inhabiting the wadis close to St Katherine in the mountains of the Sinai peninsula, were trapped and their helminth parasites were studied. Sixty one mice provided faeces for analysis and 27 were killed and autopsied. Six species of helminths were recorded (the spirurid nematodes, Protospirura muricola (74.1%) and Mastophorus muris (11.1%), the oxyuroid nematodes, Dentostomella kuntzi (59.3%), Aspiculuris africana (3.7%), and Syphacia minuta (3.7%) and the hymenolepidid cestode Rodentolepis negevi (18.5%)). The spirurids were the dominant species present, accounting for up to 0.87% of total host body weight. Analysis of worm weights and lengths suggested that transmission had been taking place in the months preceding our study. No sex difference in the prevalence or abundance of spirurids was detected. Significant differences were identified in the abundance of total nematode burdens and the mean helminth species richness between the three wadis which provided multiple captures of mice. There was also a marked effect of host age on both parameters. A highly significant positive correlation between spirurid egg counts and total worm biomass indicated that non-invasive techniques based on egg counts could be used to quantify worm burdens and when this technique was applied to a larger sample size (n = 61), a significant difference between sites but no host sex or age effects were detected for spirurid faecal egg counts. The data suggest that there are differences between helminth component communities infecting spiny mice in different neighbouring wadis, a hypothesis which will be explored further through our continuing studies in the Sinai. PMID- 10831052 TI - Prevalence and distribution of pig helminths in the Dongting Lake Region (Hunan Province) of the People's Republic of China. AB - The prevalence of helminths in pigs was investigated in five rural communities situated on the embankment of Dongting Lake in Zhiyang County, Hunan Province, People's Republic of China, in an area known to be endemic for Schistosoma japonicum. The helminth prevalences identified on the basis of faecal egg count analysis were: Oesophagostomum spp. (86.7%), Ascaris suum (36.7%), Metastrongylus spp. (25.8%), Strongyloides spp. (25.8%), Trichuris suis (15.8%), Globocephalus spp. (6.7%), Gnathostoma spp. (4.2%), Schistosoma japonicum (5.0%) and Fasciola spp. (1.3%). Post mortem examinations of a small number of pigs depositing eggs of different helminth species revealed the presence of Oesophagostomum dentatum, O. quadrispinulatum, A. suum, Metastrongylus apri, M. pudendotectus, T. suis, G. hispidum and Ascarops dentata. Prevalences of all helminths, with the exception of Oesophagostomum spp., were higher in young pigs (< 8 months old) compared with adult pigs. Prevalences of trematodes were very low, especially for S. japonicum which had decreased dramatically compared with previous reports from this area of P.R. China, whereas prevalences of nematodes were generally in agreement with those reported from other Yangtze River Provinces. Results from helminth prevalence studies in pigs, conducted in other provinces of P.R. China between 1987 and 1997, are presented and discussed. It was concluded that a government helminth control programme, implemented in 1995 to control S. japonicum infection in pigs in Hunan Province, may have resulted in a greatly reduced prevalence of S. japonicum in pigs in this region. PMID- 10831053 TI - Pomphorhynchus heronensis and restricted movement of Lutjanus carponotatus on the Great Barrier Reef. AB - Samples of Lutjanus carponotatus (Lutjanidae) from reef flat (shallow) and reef slope (deep) sites around Heron and Wistari reefs on the southern Great Barrier Reef were examined for Pomphorhynchus heronensis (Acanthocephala). Individual fish from the reef slope had 0-9 (2.6) worms as compared with 1-122 (39.6) worms for individuals from the reef flat (P < 0.0001). Other variables (year, season, size of fish) made little contribution to the variation. Reef flat and reef slope sites were separated by as little as 300 m. These results imply both that the fish have very limited local movement and that transmission of the parasite is concentrated locally. PMID- 10831054 TI - Ultrastructure of the tegument of the metacercaria of Timoniella imbutiforme. AB - The spinous body tegument of the metacercaria of Timoniella imbutiforme (Molin, 1859) is described in detail and found to comprise an outer tegumental syncytium connected to subjacently situated subtegumentary 'cells'. There are four types of secretory bodies in the outer syncytial layer as well as serrated overlapping spines and mitochondria. The subtegumentary 'cells' are characterized by the presence of four secretory body types as well as giant bodies which may be involved in the elaboration of the secretory bodies or spine material. The normal configuration of the somatic muscles of T. imbutiforme show that the muscular machinery necessary for activity once the larval stage becomes excysted is already in place. The sensory endings are found to be of the uniciliate type occurring in groups of up to eight in raised domes distributed over the body. Secretory gland cells are numerous and possess long ducts connected to the basal lamina of the outer syncytial layer via septate desmosomes. PMID- 10831055 TI - Regional specializations of the tegument of the metacercaria of Timoniella imbutiforme. AB - The ultrastructure of the spinous body tegument of the metacercaria of Timoniella imbutiforme (Molin, 1859) has recently been described. Other regions of the metacercarial tegument, including those of the oral sucker, pharynx, and nephridiopore, demonstrate considerable specializations. The oral sucker tegument had an aspinous outer syncytial layer that possessed a pimpled apical surface as well as enclosing two types of secretory bodies. The pharyngeal tegument likewise lacked spines, but possessed only one type of secretory body, and a smooth but folded outer surface. The nephridiopore tegument, however, showed the greatest degree of specialization possessing a single type of secretory body specific only to this region of the tegument. Also associated with the syncytium here was a prominent long filamentous glycocalyx, and microtubules which were observed for the first time in this region of the tegument. PMID- 10831056 TI - Ultrastructural study of spermiogenesis and the spermatozoon in Catenotaenia pusilla, an intestinal parasite of Mus musculus. AB - The ultrastructure of spermiogenesis and the mature spermatozoon in Catenotaenia pusilla (Cestoda: Catenotaeniidae) is described. Spermiogenesis is characterized by the presence of a single axoneme which grows on the outside of a cytoplasmic extension at an angle of 45 degrees. Flagellar rotation and proximodistal fusion are produced in this process. The centrioles lack striated roots and an intercentriolar body. In the mature spermatozoon four different regions are described. The anterior extremity is capped by an apical cone and presents two helical crest-like bodies of unequal length. The axoneme, of the 9 + '1' pattern of the Trepaxonemata, presents a periaxonemal sheath. The cortical microtubules form a spiral pattern at an angle of about 40 degrees to the hypothetical spermatozoon axis. The nucleus is kidney- to horseshoe-shaped in cross section. Granules and proteinaceus walls are not observed in the spermatozoon of C. pusilla. PMID- 10831057 TI - Epidemiology of Echinococcus granulosus in Arbil province, northern Iraq, 1990 1998. AB - During the period 1990-1998, 99 cases of human cystic hydatidosis (12.4 cases per year) were surgically treated at the two main hospitals in Arbil province, northern Iraq, and from this the human occurence for the province was estimated to be 2 per 100,000 inhabitants. In the same area, 1270 sheep, 550 goats and 320 cattle were examined at slaughter for hydatid cysts and prevalence rates were found to be 15.0%, 6.2% and 10.9%, respectively. A decreasing tendency in livestock prevalences was found towards the end of the study period. As in humans, most of the hydatid cysts in livestock were located in the liver. Fertility of sheep cysts, i.e. those containing protoscoleces, was found to be significantly higher (64%) than that of goats (35.7%) and cattle (29.8%). The percentage of fertile cysts containing viable protoscoleces varied between 63 and 82% in the livers and between 72 and 79% in the lungs of the different animal species. A total of 97 stray dogs were examined post-mortem in the years 1991, 1992 and 1998, and Echinococcus granulosus worms were found in the intestines of 48 dogs (49.5%). High worm burdens (> 1000) were observed in 37% of the dogs, medium worm burdens (200-1000) in 41%, and low worm burdens (< 200) in 22%. In 1998, the prevalence of canine echinococcosis (24.3%) was found to be significantly lower than in 1991 (70.4%) and 1992 (60.6%). The prevalence of human hydatidosis did not differ significantly over the years, but the study confirmed that hydatidosis is endemic in northern Iraq, and that housewives, labourers and farmers appear to be at the greatest risk of infection. PMID- 10831058 TI - Nematode parasite control practices of sheep and goat farmers in the region of Trikala, Greece. AB - Information concerning worm control practices of sheep and goat farmers in the region of Trikala (central Greece) was collected through a questionnaire survey by visiting farms and interviewing farmers. Questionnaires from 57 farmers residing in 23 rural communities were collected. Anthelmintics were used by 89% of the farmers. On average, lambs, kids and goats were treated once annually, while sheep were treated either once or twice annually. Only 2% of farmers reported treatment of animals with anthelmintics when moving to new pastures. The most common broad-spectrum anthelmintics used were those belonging to the benzimidazoles and probenzimidazoles. Fifty nine percent of the farmers used the same anthelmintic group for 3 or more years and 34% used two or more anthelmintic groups in the same year. Almost all farmers reported estimating live weights for calculating anthelmintic doses through visual perception on the basis of an average weight (96%). Tablets and boluses were the most preferred anthelmintic formulation used by 96% of farmers. The selection of an anthelmintic was based for 58% of farmers on recommendation by a veterinarian and for 39% of farmers on the cost of the drug. The most common occasions for deworming the animals were at turn out (86%) and after parturition (31%). Only 6% of farmers reported deworming new animals before introducing them onto the farm. Farmers preferred to seek information about the use of anthelmintics and worm control strategies from veterinarians (63%) and other farmers (37%). PMID- 10831059 TI - Rheological properties of fetal red cells with special reference to aggregability and disaggregability analyzed by light transmission and laser backscattering techniques. AB - Blood viscosity factors and fetal erythrocyte aggregability were investigated with light transmission (Myrenne device) during a cross-sectional study of blood drawn in utero by cord venepunctures in 119 normal fetuses between 18 and 39 weeks gestation. There was a progressive increased blood viscosity at native hematocrit (p < 0.01) explained by a gradual increase in both hematocrit (from 33% to 40%, p < 0.05) and Dintenfass' 'Tk' RBC rigidity index (p < 0.05), while plasma viscosity remained constant at 1.18 +/- 0.01 mPa x s as well as the h/eta ratio (188.4 +/- 2.7 mPa(-1) x s(-1)). The RBC aggregation index 'M' remained almost equal to zero (mean value: 0.04 +/- 0.01) before 32 wk gestation and then increased (p < 0.05) until delivery. The upper physiological limit for this parameter before 32 wk (mean +/- 2 SD) is 0.18. The RBC aggregation index 'M1' remained constant during pregnancy at 2.98 +/- 0.26, i.e., the upper physiological limit for this parameter during the intrauterine life (mean +/- 2 SD) is 7.85. Both fibrinogen (r = 0.479, p < 0.05) and albumin (r = 0.494, p < 0.01) correlated with time so that the albumin/fibrinogen ratio remained stable. We then studied with the laser retrodiffusion technique the venous blood of 20 women (18-43 yr, 37-40 wk gestation) and the cord blood of their newborns at birth, comparing RBC aggregation of: mothers (M), maternal RBCs resuspended on newborn plasma (MF), newborn RBCs resuspended on maternal plasma (FM), and newborns (F). Aggregability is higher in M (RBC aggregation time M < MF < FM < F; p < 0.01); RBC aggregation index at 10 s M > MF > FM > F; p < 0.01), with in turn the symmetric inverse picture for the partial disaggregation threshold (M > MF = FM > F). Thus RBC disaggregability is higher in newborns, and suspensions on maternal and newborn plasma suggest that half of this difference in aggregability (and disaggregability) between fetal and adult blood results from plasma factors and another half from erythrocytes. PMID- 10831060 TI - Aggregability and disaggregability of erythrocytes in women suffering from ovarian cancer: evidence for an increased disaggregation threshold. AB - In order to further characterize the alterations of erythrocyte aggregation described in ovarian cancer, we measured it with laser backscattering in eleven women suffering from ovarian cancer (mean age: 44.7 +/- 3.6, extreme values: 28 61 yr) compared with thirteen matched control women. Blood rheology exhibited a wide variability in cancer patients, with some unusually high values of plasma viscosity and/or RBC aggregation in individual cases. The only significant differences were found for the RBC disaggregation threshold which was higher in patients than in controls (78.06 +/- 10.14 vs 52.6 +/- 3.15 s(-1), p < 0.05), while hematocrit was lower (34.45 +/- 1.42 vs 38.23 +/- 0.75, p < 0.05). A negative correlation between hematocrit and corrected blood viscosity on the whole sample of subjects (r = 0.454, p < 0.05) indicates that hematocrit is decreased in subjects prone to high viscosity, resulting in similar values of apparent blood viscosity in controls and patients. Thus, a lower disaggregability of RBCs is evidenced in women with ovarian cancer, as well as a tendency to blood hyperviscosity compensated by a reduction of hematocrit which suggests that there may be some degree of 'viscoregulation'. PMID- 10831061 TI - 100 mg acetylsalicylic acid acutely decreases red cell aggregation in women taking oral contraceptives. AB - Since oral contraceptives (OC) are known to impair blood fluidity and to increase the risk of venous and arterial thrombosis, while acetylsalicylic acid (ASA) decreases the thrombotic risk and modifies some rheologic parameters, we compared the hemorheologic effects of ASA on blood rheology between women treated by OC and women who never received this medication. 25 women under OC were compared to 25 matched women who had never used OC. Blood viscosity (MT90 viscometer) and RBC aggregation (Myrenne aggregometer and AFFIBIO erythroaggregometer) were measured before and 1 hr after women received per os 100 mg ASA, after an overnight fast. The only significant difference between women under OC and controls was an increased RBC aggregation ('M' index +28%, p < 0.04; Affibio aggregation time 21%, p < 0.03). On the whole sample of 50 women as well as in the subgroup of women under OC, ASA decreased RBC partial disaggregation threshold (-1.7%, p < 0.01). These results confirm that RBC aggregation is increased under OC and suggest that 100 mg ASA acutely induces a partial reversal of this RBC hyperaggregation. PMID- 10831062 TI - Association between blood rheology, thrombosis and cancer survival in patients with gynecologic malignancy. AB - In cancer patients impaired blood rheology in the presence of coagulation activation may reduce blood flow in the vascular microcirculation that favors thrombosis but may also support tumor progression and metastasis. In 451 patients with gynecological cancer and 177 patients with corresponding benign tumor disease preoperatively, during adjuvant treatment, when venous thrombosis (VT) or cancer progression was diagnosed hematocrit (micro centrifuge), hemoglobin, leukocytes, platelets (Coulter Counter); red blood cell (RBC) aggregation (aggr.) during stasis and low shear conditions (MA 1, Myrenne), plasma viscosity (viscosimeter KSPV 1 Fresenius), and fibrinogen (Multifibren Behring Dade) were investigated. One hundred and twelve healthy women served as controls. Preoperatively, mean plasma viscosity (pv) was significantly higher in cancer patients as compared to patients with the corresponding benign tumor disease (breast cancer: n = 261; pv = 1.32 vs. 1.27 mPa s; p = 0.023; ovarian cancer: n = 68; pv = 1.39 vs. 1.31 mPa s; p < 0.001; endometrial cancer: n = 70; pv = 1.37 vs. 1.25 mPa s; p < 0.001; cervical cancer: n = 52; pv = 1.33 vs. 1.26 mPa s; p = 0.004). RBC aggr. was significantly lower in controls compared to the preoperative values in cancer patients but mean (median) values (RBC aggr. stasis < 21) were within the normal range in all. Preoperatively, plasma viscosity was a significant risk factor for the overall survival in ovarian cancer patients (p = 0.02) and for subsequent thrombosis in ovarian (p = 0.02) and cervical cancer patients (p = 0.007). In the multivariate analysis plasma viscosity was an independent prognostic marker for the overall survival of breast cancer patients (r = 99.45; 95% CI: 7.32-980.2; p < 0.0001). An optimized preoperative cut-off value above 1.40 mPa s (Log-Rank-test) was significantly associated with poor outcome in the Kaplan-Mayer survival estimates, even in node-negative breast cancer. In gynecologic cancer patients the combination of an increase in RBC aggregation and plasma viscosity impairs blood-flow-properties and may induce hypoxia in the microcirculation that favors thrombosis, settlement of tumor-cells and thus metastasis. Improvement of blood fluidity and thus oxygen transfer in the tumor-vascular-microcirculation may increase susceptibility of systemic anti cancer therapy. PMID- 10831063 TI - Decreased deformability of the X-ray-irradiated red blood cells stored in mannitol-adenine-phosphate medium. AB - X-ray irradiation of blood is an effective way to prevent transfusion-associated graft-versus-host disease. Red blood cells (RBCs) from normal donors suspended in mannitol-adenine-phosphate (MAP) medium were irradiated with X-ray of 15 and 35 Gy in minimum dose. The change of deformability of the RBCs during storage at 4 degrees C for 4 weeks was examined under shear stress of 13-130 dyn/cm2 using a rheoscope, in relation to the hematological and biochemical properties. (1) The deformability of RBCs was decreased during the storage, and it was further decreased by the irradiation. In addition, the number of undeformable RBCs against a given shear stress increased after the irradiation. (2) The cell volume gradually decreased, while the intracellular hemoglobin concentration increased. These changes were accelerated by the irradiation. The echinocytic transformation during the storage was not accelerated by the irradiation. (3) The content of aggregated proteins reducible with beta-mercaptoethanol in RBC membrane increased during the storage, but was not increased by the irradiation. Membrane lipid peroxidation was not increased during the storage and by the irradiation. (4) Leakage of potassium ions from RBCs during the storage was accelerated by the irradiation. In conclusion, shear-induced deformation of RBCs stored in MAP medium was impaired by X-ray irradiation, mainly due to dehydration caused by excess leakage of potassium ions from RBCs. PMID- 10831064 TI - The effect of low-molecular weight dextran on erythrocyte aggregation in normal and preeclamptic pregnancy. AB - Erythrocyte aggregation was determined by a novel method enabling the quantification of the aggregation process in whole blood. Blood samples of 47 healthy pregnant women and 39 preeclamptic patients were examined. Subjects within each group were matched for the gestational age. It was found that RBC aggregation increases with the gestational age in healthy pregnancy and further increases in preeclampsia. Addition of low-molecular weight dextran (MW = 9300) to blood samples of both healthy pregnant women and preeclamptic patients reduces RBC aggregation in a concentration-dependent manner. The obtained results indicate alterations in plasma composition as the primary factor for the increased RBC aggregation in both normal and pathological pregnancy. It is suggested that adsorption of low-molecular weight dextran on the RBC membrane reduces the surface concentration of plasma bridging molecules thereby reducing RBC aggregation toward normal. PMID- 10831065 TI - Modifications of whole blood filterability during acute myocardial infarction. AB - Experimental evidences underline that hemorheological alterations observed in acute myocardial infarction (AMI) are strictly involved in the decreased perfusion of the damaged area and in the extension of the necrotic regions. We have analyzed whole blood filterability as an index of erythrocyte deformability in 60 AMI patients compared with 30 patients with non-acute coronary artery disease and 52 healthy subjects. Nucleopore polycarbonate membranes with a pore diameter of 5 microm and a filtering pressure of -20 cm H2O were used. The results are expressed as the volume of whole blood filtered in 1 minute (index of filterability, IF). In normal subjects IF was 1.16 +/- 0.24. Among AMI patients IF was 0.70 +/- 0.30 at admission, 0.68 +/- 017 at day 10 and 0.78 +/- 0.14 at day 20. These values were significantly lower than those obtained in normal subjects and in patients with non-acute coronary artery disease. In addition, AMI patients treated with thrombolytic therapy showed, at admission, a significantly higher IF value than that obtained in patients who did not receive thrombolytic treatment (0.85 +/- 0.34 vs 0.60 +/- 0.22; p < 0.01). These results demonstrate an evident reduction of whole blood filterability in AMI patients that may be considered as an index of erythrocyte deformability. Thrombolytic therapy seems to have a positive effect on blood filterability and may produce beneficial effects through its therapeutical action other than the lysis of the coronary thrombus. PMID- 10831066 TI - Standard aggregating media to test the "aggregability" of rat red blood cells. AB - Rat red blood cells (RBC) exhibit low aggregation tendency in autologous plasma and in standard aggregating media (e.g., 3% Dextran 70; MW: 70 kD). In experimental studies performed on rats, 3% Dextran 70 was found to be an unsuitable suspending medium to test the "aggregability" of RBC in a standard medium. It has been observed that solutions of higher molecular weight polymers (i.e., dextran, MW: 500 kD; polyethylene glycol , MW: 35 kD; polyvinylpyrrolidone, MW: 360 kD), at low concentrations were strong aggregators for rat RBC. Among these polymer solutions 0.5% Dextran 500 and 0.75% polyvinylpyrrolidone 360 were found to be the most suitable suspending media, to distinguish between the aggregability of RBC from septic and control rats. Therefore, these two polymer solutions are recommended as the standard aggregating media for rat RBC, to test the RBC aggregability. PMID- 10831067 TI - Ligustrazine, allicin and shear-induced platelet aggregation. PMID- 10831068 TI - Hemorheology in microcirculation: pathological changes--Internet/E-mail discussion proceeding from October 1998 to June 1999. Report on the 7th Tbilisi Symposium. PMID- 10831069 TI - Role of growth factors in osteoblast alterations induced by skeletal unloading in rats. PMID- 10831070 TI - Stoichiometry, kinetic and binding analysis of the interaction between epidermal growth factor (EGF) and the extracellular domain of the EGF receptor. AB - The kinetics, binding equilibria and stoichiometry of the interaction between epidermal growth factor and the soluble extracellular domain of the epidermal growth factor receptor (sEGFR), produced in CHO cells using a bioreactor, have been studied by three methods: analytical ultracentrifugation, biosensor analysis using surface plasmon resonance detection (BIAcore 2000) and fluorescence anisotropy. These studies were performed with an sEGFR preparation purified in the absence of detergent using a mild two step chromatographic procedure employing anion exchange and size exclusion HPLC. The fluorescence anisotropy and analytical ultracentrifugation data indicated a 1:1 molar binding ratio between EGF and the sEGFR. Analytical ultracentrifugation further indicated that the complex comprised 2EGF:2sEGFR, consistent with the model proposed recently by Lemmon et al. (1997). Global analysis of the BIAcore binding data showed that a simple Langmuirian interaction does not adequately describe the EGF:sEGFR interaction and that more complex interaction mechanisms are operative. Furthermore, analysis of solution binding data using either fluorescence anisotropy or the biosensor, to determine directly the concentration of free sEGFR in solution competition experiments, yielded Scatchard plots which were biphasic and Hill coefficients of less than unity. Taken together our data indicate that in solution there are two sEGFR populations; one which binds EGF with a KD of 2-20 nM and the other with a KD of 400-550 nM. PMID- 10831071 TI - Multiple autocrine factors including an extracellular matrix protein are required for the proliferation and spreading of human colon carcinoma cells in vitro. AB - The human colon carcinoma cell line LIM1215 proliferates and changes morphology (spread) in a cell density-dependent manner in response to epidermal growth factor (EGF). At high density, production of autocrine transforming growth factor alpha enables the cells to proliferate and spread in the absence of exogenous EGF or serum. At low cell density (< 1 x 10(4)/cm2) EGF alone fails to elicit a mitogenic or morphological response and requires the presence of conditioned medium (derived from high cell density serum-free culture of the same cells) to exert its effects. This synergy between EGF and LIM1215 conditioned medium was investigated further. Using a low cell density assay and fractionated LIM1215 conditioned medium, we show that EGF-mediated mitogenic and morphological responses are separable. These responses are dependent on the synergistic action of a low molecular weight autocrine survival factor and an extracellular matrix like spreading factor(s) secreted into the culture medium respectively. We find that under low cell density, serum-free conditions, EGF alone is insufficient to rescue LIM1215 from rapid apoptotic death. Catalase or LIM1215 autocrine survival factor prevent the death of LIM1215 cells and restore their proliferative (but not morphological) response to EGF, suggesting that cell death under these conditions may be the result of oxidative stress. Combination of EGF, partially purified autocrine survival and spreading factors induced proliferation and spreading of low density LIM1215 cells similar to that observed with EGF and unfractionated conditioned medium. GRGDS peptides strongly inhibited the spreading of LIM1215 cells in the presence of EGF and the partially purified autocrine spreading factor, demonstrating that integrin receptors are involved in the spreading process. Comparison of the spreading response of LIM1215 and Colo 526 cells on ASF and various adhesion proteins indicate that ASF is not collagen I, collagen-IV, fibronectin or vitronectin. Taken together, these results support the concept that the autonomous growth of colon carcinoma cells in vitro is dependent on the synergistic interaction between several autocrine systems. PMID- 10831072 TI - Tissue distribution and retinoid-mediated downregulation of an FGF-binding protein (FGF-BP) in the rat. AB - We showed previously that a secreted fibroblast growth factor-binding protein (FGF-BP) can mobilize and bioactivate locally-stored FGFs from the extracellular matrix. This FGF-BP is upregulated in various cancers and plays a rate limiting role as an angiogenic switch molecule during tumor growth. In this paper, we describe the cloning and sequence analysis of the rat homologue of FGF-BP and show its expression pattern and retinoid-mediated downregulation in normal adult rat tissues. The rat FGF-BP amino acid sequence is 91% and 70% homologous to mouse and human, respectively, and contains 10 cysteine residues whose position is conserved across species. In Northern blots, FGF-BP mRNA was detected in the gut, eye, thymus, skin, lung and tongue. Immunohistochemistry confirmed this tissue distribution with cerebellar Purkinje cells, the cerebral chorioid plexus and the eye showing the most distinctive staining patterns. Oral treatment of animals with all-trans-retinoic acid for one and two days induced a significant decrease of FGF-BP protein in tissues from stomach, eye and lung suggesting that regulation of FGF-BP can be one effector mechanism through which retinoids affect normal and pathological processes. PMID- 10831073 TI - T-cell-specific expression of kinase-defective Eph-family receptor protein, EphB6 in normal as well as transformed hematopoietic cells. AB - Although most kinase-defective growth factor receptor proteins are associated with pathogenic conditions, a kinase-defective Eph-family receptor protein, EphB6, is expressed in normal human tissues. We generated monoclonal antibodies specific for human EphB6 to characterize its expression on human hematopoietic cells. A very small population of normal human peripheral white blood cells (0.57 +/- 0.07%, n = 12) expressed EphB6. The EphB6-positive cells were CD2+, CD7+, CD3+ and CD4+ or CD8+ lymphocytes, but they did not express CD19 or CD11b. In human bone marrow, only 1.5 +/- 0.19% of lymphocytes expressed EphB6. Compared with the expression in peripheral lymphocytes, prominent expression of EphB6 protein was demonstrated in CD4+CD8+ double-positive mouse thymocytes. The T-cell lineage-specific expression was strictly conserved in human leukemia/lymphoma cells. Among T-cell-derived leukemia cells, the expression level of EphB6 seemed to decrease with maturation of the cells. These results suggest that EphB6 expression is regulated in T-cell development. PMID- 10831074 TI - Intravenous mycophenolate mofetil: safety, tolerability, and pharmacokinetics. AB - An intravenous (i.v.) formulation of mycophenolate mofetil (MMF; CellCept, Roche Pharmaceuticals, Inc., Palo Alto, CA) that will enable its administration to patients unable to tolerate oral medication is available. Two separate studies, an open-labeled pharmacokinetic (PK) study and a double-blind safety study, were performed. Within 24 h after transplant, 153 (safety study) and 45 (PK study) first or second renal transplant recipients were started on i.v. MMF 1 g Q12h or placebo (used in the safety study only, 2:1 MMF:placebo), given over 2 h via a dedicated peripheral venous catheter. In the safety study, per os (p.o.) MMF (1g Q12h) or placebo was administered, starting within 72 h after transplant, whereas in the PK study, p.o. MMF was started on the evening of day 5. Sequential blood samples obtained on study days 5 (i.v. MMF) and 6 (p.o. MMF) before and up to 12 h after the AM dose were analyzed for mycophenolic acid (MPA) and MPA glucuronide (MPAG) concentrations by high-performance liquid chromatography. The area under the concentration curve (AUC) was calculated using the linear trapezoidal rule. The MPA AUC(0-12) was higher for i.v. MMF than p.o. MMF (40.8 +/- 11.4 microg x h/ mL vs. 32.9 +/- 15, p < 0.001). There were no other significant PK differences for plasma MPA or MPAG. In the safety study (n = 98 i.v. MMF vs. n = 55 placebo), 11 patients (11%, i.v. MMF) and 4 patients (7%, placebo) discontinued their use of the drug because of an adverse event (AE). Overall, AEs were similar between i.v. MMF and placebo. Injection site phlebitis (4%) and thrombosis (4%) were observed only with i.v. MMF. MMF i.v. 1 g twice daily (b.i.d.) should provide efficacy at least equivalent to p.o. MMF without increased toxicity, and it provides an acceptable alternative dose form in the immediate period after transplant. PMID- 10831075 TI - A tale of two kidneys--how long can a kidney transplant wait? AB - This paper compares early graft function (EGF) of the first transplanted kidney (group 1) with the kidney transplanted second (group 2) in kidney pairs from the same cadaver donor. Thirty-one pairs of kidneys were harvested from cadaver donors between January 1997 and October 1998. Each pair was transplanted using a standard technique by the same team of surgeons, one after the other, as a result of limitations in theatre time and staff availability. Incidence of acute rejection (AR), acute tubular necrosis (ATN) and need for post-transplant dialysis was recorded for both groups, and was compared using the relevant statistical methods. Patients in both groups were well matched for age, gender and mode of dialysis pre-transplant. Human leucocyte antigen (HLA) matching and panel reactive antibody (PRA) status were similar in the two groups (p > 0.05). Cold ischaemia time (CIT) in the two groups was 14.1 +/- 5.7 and 19.2 +/- 6.9 h, respectively, the difference being statistically significant (p < 0.05). The incidence of AR was similar in the two groups. However, ATN (on renogram) was significantly more common in group 2 (p < 0.05; 12 patients versus 5 patients in group 1). All patients with ATN required post-transplant dialysis. Hospital stay was significantly prolonged in group 2 patients (p < 05; 20 +/- 10.6 versus 16.3 + 6.2 d for group 1). Even a relatively short increase in CIT can cause the second transplanted kidney of a pair to have a significantly higher incidence of ATN, resulting in need for dialysis and prolongation of hospital stay. Simultaneous transplantation, in areas lacking organ sharing networks, would not only improve EGF, but also improve long term graft survival. In addition, the reduced requirement for post-transplant dialysis and a shorter hospital stay would balance any increased demand on resources. PMID- 10831076 TI - Failure of ganciclovir prophylaxis to completely eradicate CMV disease in renal transplant recipients treated with intense anti-rejection immunotherapy. AB - Ganciclovir prophylactic regimens have been shown to be effective in renal transplant recipients at risk for primary (donor seropositive/recipient seronegative) and secondary (recipient seropositive) cytomegalovirus (CMV) disease. However, in addition to serologic factors, the type and intensity of the administered immunosuppression is a strong risk factor for CMV disease. Since January 1995, we have utilized a potent immunosuppressive protocol selectively in recipients at high risk for immunologic graft loss, defined as retransplant recipients, recipients with delayed graft function, non-Caucasian recipients, and recipients suffering from acute rejection. Between January 1995 and December 1996, 110 consecutive renal transplants were performed in recipients who were either CMV seropositive or received an allograft from a CMV-seropositive donor. All recipients received ganciclovir prophylactic therapy for 3 months post transplant. Group I (N = 43) consisted of recipients at high-immunologic risk for graft loss as defined above. These recipients were treated with an intense anti rejection immunotherapeutic regimen consisting of Cellcept, Neoral, and prednisone, with the frequent addition of antilymphocyte antibody therapies and intravenous methylprednisolone. The remaining 67 recipients (group II) were treated with a less intense immunotherapeutic regimen consisting of azathioprine, Neoral, and prednisone. The incidence and severity of CMV disease and the patient and allograft survival were compared. The incidence of CMV syndrome was greater in group I (28%) compared with group II (7%), and was statistically significant (p < 0.05). The 1-yr patient and graft survival were similar, 95 and 91%, respectively, for group I compared with 97 and 97%, respectively, for group II. These data suggest that 3 months of ganciclovir prophylactic therapy is significantly less effective for the prevention of CMV disease in renal transplant recipients at high risk for acute rejection treated with an intense immunotherapeutic regimen. These data suggest that more effective prevention of CMV disease in these high-risk recipients will require the addition of other anti viral agents, such as immunoglobulin preparation to the prophylactic regimen. PMID- 10831077 TI - Health related quality of life (HRQOL) of kidney transplanted patients: variables that influence it. AB - The incidence and prevalence of patients on renal replacement therapy (RRT) who receive a renal transplant are continuously increasing in Spain. At the moment, they are the main group of end-stage renal disease (ESRD) patients in our region (60% of total RRT patients). The aim of the present study was to assess the health related quality of life (HRQOL) of kidney transplanted patients of our region, and to identify socio-demographic and clinical variables that influence it. The intention was also to compare the HRQOL of these patients with that of chronic haemodialysis (HD) patients and of the general population. METHODS: Two hundred and ten kidney transplanted patients and 170 HD patients were evaluated using the Karnofsky performance scale (KPS), sickness impact profile (SIP), and SF-36 Health Survey (SF-36). Socio-demographic and clinical data, including a comorbidity index (CI), were also collected. To compare our patients with the general population we used SF-36 mean scores from an aleatory sample taken from our region. RESULTS: Transplant patients had lower mean scores on SIP dimensions and higher scores on SF-36 dimensions than chronic HD patients. In transplant patients, we found significant differences on SIP and SF-36 scores in gender, educational level, haematocrite and haemoglobin, CI, time since transplantation, and KPS. CONCLUSIONS: The HRQOL of transplant patients is clearly better than that of chronic HD patients and similar to that of the general population. Differences in the HRQOL within transplant patients did not appear to be as a result of patient's age, but rather it would appear to be a consequence of gender, analytic figures, CI, KPS score, time with transplant, and educational level. PMID- 10831078 TI - Levels of HBV-DNA and HBsAg after acute liver allograft rejection treatment by corticoids and OKT3. AB - The aim of this work was to analyze whether the treatment of acute rejection of orthotopic liver transplants (OLT), either with corticoids or OKT3, has any effect on the levels of hepatitis B virus (HBV)-DNA and HBsAg in individuals which were originally affected by cirrhosis or fulminant hepatic failure as a result of B virus. We have found that HBV-DNA is present in macrophages, B cells and both CD4+ and CD8+ T cells after OLT in all cases studied. Interestingly, the levels of HBV-DNA and HBsAg in the serum analyzed were increased extremely rapidly in the patients treated with OKT3 in an acute rejection episode. However, the serum levels of HBV-DNA and HBsAg found were lower when the patients were treated with steroids, and were not found in non-treated patients. As the serum levels of HBV-DNA increase, the process of liver reinfection could be accelerated; therefore, these results may help to understand how OKT3 and corticoids immunosuppressive therapy may accelerate the reinfection of OLT by HBV. In conclusion, our results suggest that special care must be taken in the use of OKT3 in the treatment of acute liver rejection episodes in chronic or fulminant HBV transplanted patients. PMID- 10831079 TI - Effect of recipient sensitization (peak PRA) on graft outcome in haploidentical living related kidney transplants. AB - OBJECTIVE: To evaluate the influence of pre-transplant recipient sensitization on the outcome of 1-haploidentical live related donor (LRD) kidney transplants. METHOD: We reviewed 141 consecutive cyclosporine-treated adult haploidentical first transplants for which panel reactive antibody (PRA) levels were available. Patients were divided into three groups according to their peak PRA levels: group I, PRA = 0 (n = 97); group II, PRA = 1-50% (n = 24); and group III, PRA = 51-100% (n = 20). RESULTS: Differences in PRA were associated with significant differences in short- and longer-term graft survival, unrelated to patient survival. Graft survival at 1, 3, and 5 yr was only 74, 40, and 27% in group III, compared to 92, 87, and 52% in group II, and 96, 91, and 85% in group I (p < 0.001). Increasing PRA was associated with shorter time-to-graft failure. In group III, 20% lost their transplant from acute rejection in the first 6 months, versus 4% in group II and 3% in group I (p < 0.01). Graft survival in group II diverged from that of group I only after 3 yr, due to an increase in loss from chronic rejection. Hospitalization was longer in group III, in association with a significantly higher incidence of acute rejection during the first 3 months after transplantation (p < 0.02). Serum creatinine was higher in sensitized than nonsensitized patients at all time points. CONCLUSIONS: Sensitization has a significant negative impact on the outcome of haploidentical LRD kidney transplants. Sensitized potential recipients and their potential donors should be aware of this in arriving at informed decision-making for transplantation. These patients may benefit from more sensitive cross-match testing, more intense or more novel immunosuppression, or immunomodulation to modify their immune responsiveness. PMID- 10831080 TI - Peripheral endothelial dysfunction in heart transplant recipients: possible role of proinflammatory cytokines. AB - Endothelium-dependent vasodilation in the peripheral circulation may be impaired in heart transplant recipients (HTx rec). Conflicting results have been obtained and the mechanisms involved have not been examined. In the present study, we examined whether long-time survivors of heart transplantation (Tx) show signs of endothelial dysfunction in the peripheral microcirculation, and further investigated the possible role of endothelium-related markers and proinflammatory cytokines in this process. The vasodilatory responses to acetylcholine (Ach) (endothelium-dependent) and sodium nitroprusside (SNP) (endothelium-independent) were evaluated by skin laser-Doppler perfusion measurements in 63 clinically stable HTx rec 6 yr (range 1-13 yr) after Tx, and compared with 20 healthy controls. Ten HTx rec were also followed prospectively with three repeated measurements during the first year after Tx. Plasma von Willebrand factor, big endothelin (b-ET), and proinflammatory cytokines were measured by enzyme immunoassays. Vascular responses to both Ach and SNP were significantly attenuated in the HTx rec compared with controls. In longitudinal testing, there was a significant reduction in endothelium-dependent vasodilation, but not independent vasodilation from 1 to 12 months after Tx. Plasma levels of vWF and b ET, as well as levels of proinflammatory cytokines, tumor necrosis factor (TNF) alpha, interleukin (IL)-6 and IL-1beta, were all markedly increased in HTx rec. HTx rec responses to Ach were negatively correlated to TNF-alpha levels in plasma (r = -0.39, p < 0.01). Moreover, there was also a significant positive correlation between plasma b-ET and TNF-alpha (r = 0.34, p < 0.01). In the long term follow-up of HTx rec, endothelial dysfunction is demonstrated by both regulation of blood flow in the skin microcirculation and by raised markers of endothelial activation in plasma. This endothelial dysfunction may be related to enhanced levels of proinflammatory cytokines in these patients. PMID- 10831081 TI - Renal transplantation for type II diabetic patients compared with type I diabetic patients and patients over 50 years old: a single-center experience. AB - INTRODUCTION: Transplantation of renal allografts for Type II diabetic patients has become increasingly common. Like Type I diabetics and patients over age 50, Type II diabetics are in a high-risk category for renal transplantation. Some investigators argue that the scarce resources of the valuable donor pool should not be allocated to these high-risk, older individuals with end-stage renal disease (ESRD) who often have significant comorbidity. We studied the outcome of renal transplantation for 90 Type II diabetics and compared it with the outcome for Type I diabetics and patients over age 50 with other primary diseases. METHODS: We conducted a retrospective review of data on all renal transplants performed at the University of Minnesota since September 1984 for Type II diabetics. We analyzed both patient and graft survival rates and compared them with the outcome for Type I diabetics and separately for all nondiabetics over age 50. We grouped recipients by donor source (living vs. cadaver) and age at transplant (< 50 vs. > 50). We studied reasons for graft loss, patient death, pre operative morbidity, and post-operative complications. RESULTS: We found that recipient age and donor source did not affect outcome. Overall 5-yr patient survival for Type II diabetics was 61%; graft survival, 53%. Type II diabetics had significantly worse patient and graft survival than Type I diabetics and than nondiabetics over age 50. However, death-censored graft survival was not different between these groups, suggesting little difference in immunologic graft loss. CONCLUSIONS: Renal transplantation is a relatively safe, viable option for Type II diabetics with ESRD and significant comorbidity. Although their overall graft survival is less than for Type I diabetics and for nondiabetics over age 50, the immunologic graft failure rate is the same in all three groups. PMID- 10831082 TI - Venous complications after orthotopic liver transplantation. AB - Complications involving the portal vein or the vena cava, are rare after orthotopic liver transplantation. We report on the incidence and treatment of venous complications following 1000 orthotopic liver transplantations in 911 patients. Twenty-six of the adult patients (2.7%) suffered from portal complications after transplantation, whereas complications of the vena cava were observed in only 17 patients (1.8%). Technical problems or recurrence of the underlying disease (e.g. Budd-Chiari syndrome) accounted for the majority of complications of the vena cava, whereas alteration of the vessel wall or splenectomy during transplantation could be identified as important risk factors for portal vein complications. In patients undergoing modification of the standard end-to-end veno-venous anastomosis of the portal vein due to pathological changes of the vessel wall, complications occurred in 8.3%, whereas only 2.4% of patients who received a standard anastomosis of the portal vein experienced complications of the portal vein. Furthermore, splenectomy during transplantation was also associated with an increased incidence of portal vein complications (10.5 vs. 2.2% in patients without splenectomy). Treatment was dependent on the signs and symptoms of the patients, and varied considerably between patients with portal vein complications and patients suffering from complications of the vena cava. Complications of the vena cava led to retransplantation in about one-third of the patients, whereas in patients with occlusion of the portal vein, retransplantation was necessary in only 15%, and more than half of the patients suffering from portal vein complications did not require any treatment at all. Usually, treatment of patients with portal vein complications only became necessary when additional complications such as arterial occlusion or bile duct injuries occurred. PMID- 10831083 TI - Towards a specific immunosuppression for pancreas and islet grafts. AB - This brief article is a summary of the presentations at a meeting organized in Lyon, France, to discuss strategies of reducing post-transplant diabetes mellitus and to suggest an ideal immunosuppressive therapy for pancreatic and islet allografts. PMID- 10831084 TI - Scanning electron microscopic analysis of endothelial cell coverage and quality in large vessels from multi-organ donors: effects of preservation on endothelial cell integrity. AB - Endothelial cell integrity (coverage and quality) of large donor vessels is important because these vessels are used for vascular reconstructions in solid organ transplantation. Disruption of the endothelial cell monolayer will initiate blood coagulation and may lead to thrombosis of large vessels, often resulting in the loss of the transplanted organ. Iliac arteries and veins, removed from 10 heart-beating multi-organ donors at the end of the donor procedure, were analyzed using scanning electron microscopy at three different time points of preservation. Endothelial cell coverage and quality were determined immediately after removal from the donor, after 10 h (time of transplantation) and 7 d storage in 'University of Wisconsin' cold preservation solution (UW). Endothelial cell coverage decreased during the preservation of arteries, but was maintained in veins. Storage of the veins for 7 d in plastic bags showed a decreased endothelial cell coverage compared to storage in glass vials. Early removal of the blood vessels and proper storage, free floating and in clean UW, may improve maintenance of the endothelial cell integrity. These findings may be important in order to reduce the risk of thrombosis and, consequently, organ failure after transplantation. Furthermore, vessels with maintained endothelial cell integrity after 7 d may be used for in vitro research. PMID- 10831085 TI - Students' acceptance of clinical xenotransplantation. AB - The aim of this study was to elucidate undergraduate university students' views on clinical xenotransplantation. A total of 1875 students from eight faculties at Uppsala University and the Swedish University of Agricultural Sciences answered a questionnaire. Three out of four respondents would be prepared to receive a transplant from an animal on medical grounds if necessary. Forty percent had signed an organ donation card. There was no difference in attitude between those who had signed an allotransplantion card and those who had not. According to gender, age, length of university program, and faculty, results showed that a higher proportion of those who approved were male, young, and studying on programs longer than three years; also, they were more likely to study programs in the Faculties of Agriculture and Pharmacy. At the Medical Faculty, nursing students seemed to be less approving, compared to future biomedical analysts, biomedical scientists, and physicians. The acceptance of xenotransplantation also tended to be positively associated with morally accepting and understanding the use of animals in biomedical research, the approval of euthanasia, the approval of early abortion, and the use of human fetuses in research, as well as clinical testing of humans. PMID- 10831086 TI - The beneficial effects of calcium channel blockers on long-term kidney transplant survival are independent of blood-pressure reduction. AB - There is a growing body of evidence suggesting that calcium channel blockers (CCB) exert beneficial effects on kidney transplant survival. However, it is not completely understood if these agents act independently of blood-pressure reduction. In the present study, the 5-yr follow-up of 45 kidney transplant recipients receiving CCB during the 60-month follow-up period was compared to that of recipients with lower blood pressure and an antihypertensive treatment without CCB. During the whole follow-up, systolic (127.4 +/- 2.5 vs. 139.4 +/- 2.1 mmHg, p < 0.05) as well as diastolic blood pressure (78.8 +/- 1.1 vs. 84.8 +/ 1.8 mmHg, p < 0.05) was higher in the group receiving CCB. Moreover, in CCB treated recipients, a significant (p < 0.05) higher increase in proteinuria was detected (from 759 +/- 120 to 1690 +/- 359 mg/24 h vs. 180 +/- 45 to 340 +/- 45 mg/24 h). Despite higher blood pressure and higher proteinuria, the increase in serum creatinine in the group of CCB-treated recipients was significantly lower (0.01 mg/dL/month) in comparison to that of the controls (0.02 mg/dL/month, p < 0.05). Moreover, the 5-yr transplant survival was significantly higher in CCB treated recipients (62.3 vs. 31.8%, p < 0.05). The results of the present study further support the beneficial effects of CCB in kidney transplant recipients, which are independent of blood-pressure reduction. PMID- 10831087 TI - Tacrolimus-induced hemolytic uremic syndrome and end-stage renal failure after liver transplantation. AB - BACKGROUND: Hemolytic uremic syndrome (HUS) is a rare complication in solid organ transplantation. It can be associated with severe hypertension. Several risk factors have been identified including immunosuppressive drugs such as cyclosporin A and, more recently, tacrolimus. METHODS: Here we report a case of tacrolimus-induced HUS in a 61-yr-old woman after liver transplantation. Hypertension, microangiopathic anemia and end-stage renal failure occurred 2 yr after liver transplantation. RESULTS: At admission, she had malignant hypertension with a severe hypertensive retinopathy, renal failure (creatininemia: 800 micromol/L) and microangiopathic anemia (Hb: 7.3 g/dL, a low platelet count and elevated lactate dehydrogenase). At renal biopsy, histologic findings were ischemic and sclerotic glomeruli with hyaline thrombi, severe mesangiolysis and interstitial fibrosis. CONCLUSION: Despite steroid treatment, antihypertensive agents and fresh frozen plasma therapy, end-stage renal failure was observed and chronic hemodialysis treatment was required. PMID- 10831088 TI - Non-heart-beating kidney donors. PMID- 10831089 TI - Testing of dentin adhesives. PMID- 10831090 TI - International collaborative research on fluoride. PMID- 10831091 TI - MALDI-TOF MS analysis of a library of polymerizable cyclodextrin derivatives. AB - Polymerizable cyclodextrin derivatives (PCDs) have been proposed as candidates for use in dental therapeutics (Bowen, 1996; Bowen and Reed, 1997). Here, PCD "libraries" were synthesized by quasi-random reactions of 6 moles of methacrylic anhydride plus 6 moles of cyclic glutaric anhydride per mole of beta-cyclodextrin (BCD) in solution. BCD has 21 reactive sites on each of its molecules. These proportions were based on probability calculations, which predicted that the products should have a minimum of 2 polymerizable substituents and acidic ligand groups on practically every one of the diverse product molecules. Matrix-assisted laser desorption/ionization (MALDI) time of flight (TOF) mass spectrometry (MS) gave valuable information regarding the masses of molecular ions representing the molecules that made up the PCD libraries. For the MALDI-TOF MS analyses, small samples were analyzed by the successive application of 3 solutions to the sample holder: the matrix in acetone, the products in water, and sodium trifluoroacetate in water. The resulting spectra had > 40 envelopes of mass peaks above background. The ionic-abundance peak heights had quasi-Gaussian configurations, with central peaks having masses in the neighborhood of 2000 g/mol (Daltons). Regardless of structural permutations within each peak, the range of these peaks was between about 1500 g/mol and 2900 g/mol. This range of masses was in accord with, but perhaps somewhat more narrow than, that predicted by the statistical method, which was based on equal reactivity of all hydroxyl groups. Analysis by MALDI-TOF MS gave valuable data regarding the masses, structures, and characteristics of the products formed and provided unanticipated information to facilitate improvements in future PCD syntheses. PMID- 10831092 TI - Cloning human enamelin cDNA, chromosomal localization, and analysis of expression during tooth development. AB - Enamelin is the largest protein in the enamel matrix of developing teeth. In the pig, enamelin is secreted as 186-kDa phosphorylated glycoprotein, which is rapidly processed by enamel proteinases into smaller cleavage products. During the secretory stage of enamel formation, enamelin is found among the crystallites in the rod and interrod enamel and comprises roughly 5% of total matrix protein. Although the function of enamelin is unknown, it is thought to participate in enamel crystal nucleation and extension, and the regulation of crystal habit. Here we report the results of enamelin in situ hybridization in a day 1 mouse developing incisor that shows that enamelin is expressed by ameloblasts, but not by odontoblasts or other cells in the dental pulp. The restricted pattern of enamelin expression makes the human enamelin gene a prime candidate in the etiology of amelogenesis imperfecta (AI), a genetic disease in which defects of enamel formation occur in the absence of non-dental symptoms. We have cloned and characterized a full-length human enamelin cDNA and determined by radiation hybrid mapping and fluorescent in situ hybridization (FISH) that the gene is located on chromosome 4q near the ameloblastin gene in a region previously linked to local hypoplastic AI in six families. These findings will facilitate the search for specific mutations in the enamelin gene in kindreds suffering from amelogenesis imperfecta. PMID- 10831093 TI - Inhibition of Wnt signaling by exogenous Mfrzb1 protein affects molar tooth size. AB - Wnt extracellular signaling molecules have essential roles as regulators of cell proliferation, migration, differentiation, and in epithelial-mesenchymal interactions involved in tissue morphogenesis. Frizzled integral membrane proteins have been shown to function as receptors for Wnt signaling molecules. Vertebrates also produce secreted proteins related to Frizzled receptors, Frizzled-related proteins (FRPs), which contain the cysteine-rich domain of Frizzleds and appear to function as Wnt antagonists. Tooth development is regulated by a reciprocal series of epithelial-mesenchymal interactions, and many Wnt signaling pathway genes are expressed in the developing tooth at these sites. Here we report the expression of one FRP gene, Mfrzb1, in the rostral mesenchyme of the mandibular primordium. Using explant cultures, we show that expression of Mfrzb1 in the mandibular mesenchyme is under the control of signals derived from the overlying epithelium. Bead implantation experiments in vitro show that FGF8 induces Mfrzb1 expression, whereas BMP4 and SHH proteins have no effect. We studied the effect of ectopic MFrzb1 protein on the developing tooth germs by transplanting explants treated with Mfrzb1 protein into renal capsules, and found it to retard tooth development. This suggests that Wnt signaling is required early in tooth germ formation and that interference with signaling via addition of an antagonist results in retarded development and formation of smaller teeth. PMID- 10831094 TI - Low levels of GRP-Ca expression in transgenic mice. AB - The GRP-Ca gene of the rat encodes a member of the glutamine/glutamic-acid-rich protein (GRP) family. This gene is expressed in a highly tissue-specific fashion, with transcription being found only in the acinar cells of the submandibular gland (SMG). To begin to define the cis-acting elements governing GRP-Ca expression, we constructed transgenic mice containing the rat GRP-Ca gene plus 9.5 kb of 5' genomic sequence and 1 kb of 3 genomic sequence. Expression of GRP Ca was detectable in progeny from only 1 of 3 independent founders. Expression levels of the transgenic GRP-Ca were much lower than the native GRP-Ca found in the rat SMG. Furthermore, GRP-Ca in transgenic mice was not tissue-specifically expressed, being found in both the SMG/SLG complex and the liver. These results indicate that the genomic region of GRP-Ca included in these transgenic mice is not sufficient to confer the high levels of tissue-specific expression seen in the rat. PMID- 10831095 TI - Pellicle precursor protein crosslinking characterization of an adduct between acidic proline-rich protein (PRP-1) and statherin generated by transglutaminase. AB - Recent work with oral transglutaminase indicated that this enzyme, derived from oral epithelial cells, crosslinked pellicle precursor proteins which may be important in the formation of the acquired enamel pellicle. The purpose of this study was to investigate whether purified acidic PRP-1 can form crosslinks with statherin, and whether such a crosslink is derived from a transglutaminase catalyzed reaction between glutaminyl and lysyl side-chains, leading to a covalent bond formation. Enzymatic reaction products were analyzed by SDS-PAGE and reverse-phase HPLC. The SDS electrophoretogram revealed a protein band with an apparent molecular weight of 32 kDa, which is consistent with the combined apparent molecular weight of acidic PRP-1 (24 kDa) and statherin (8 kDa). A reaction product isolated by HPLC was characterized by amino acid analysis, which showed a stoichiometry consistent with being an adduct composed of one molecule of acidic PRP-1 and one molecule of statherin. In negative control experiments, it could be shown that this adduct was not detected when the lysines of both substrates were modified by reductive methylation prior to the enzymatic reaction. In addition, amino acid analysis and mass spectrometry confirmed the presence of a gamma-glutamyl-epsilon-lysine dipeptide after enzymatic hydrolysis and the absence of this dipeptide after acid hydrolysis. Analysis of the data obtained indicates that oral transglutaminase is capable of crosslinking acidic PRP-1 and statherin in vitro. In addition, this finding exemplifies the potential of post-secretory processing of salivary proteins, which may represent an additional mechanism to generate new protein species. PMID- 10831096 TI - Choline acetyltransferase, acetylcholinesterase, and nicotinic acetylcholine receptors of human gingival and esophageal epithelia. AB - A non-neuronal cholinergic system that includes neuronal-like nicotinic acetylcholine receptors (nAChRs) has recently been described in epithelial cells that line the skin and the upper respiratory tract. Since the use of nicotine containing products is associated with morbidity in the upper digestive tract, and since nicotine may alter cellular functions directly via nAChRs, we sought to identify and characterize a non-neuronal cholinergic system in the gingival and esophageal epithelia. mRNA transcripts for alpha3, alpha5, alpha7, and beta2 nAChR subunits, choline acetyltransferase, and the asymmetric and globular forms of acetylcholinesterase were amplified from gingival keratinocytes (KC) by means of polymerase chain-reactions. These proteins were visualized in the gingival and esophageal epithelia by means of specific antibodies. Variations in distribution and intensity of immunostaining were found, indicating that the repertoire of cholinergic enzymes and receptors expressed by the cells changes during epithelial maturation, and that an upward concentration gradient of free acetylcholine exists. Blocking of the nAChRs with mecamylamine resulted in reversible loss of cell-to-cell adhesion, and shrinking and rounding of cultured gingival KC. Activation of the receptors with acetylcholine or carbachol caused stretching and peripheral ruffling of the cytoplasmic aprons, and formation of new intercellular contacts. These results demonstrate that both the keratinizing epithelium of attached gingiva and the non-keratinizing epithelium lining the upper two-thirds of the esophageal mucosa possess a non-neuronal cholinergic system. The nAChRs expressed by these epithelia are coupled to regulation of cell adhesion and motility, and may provide a target for the deleterious effects of nicotine. PMID- 10831097 TI - EMG power spectrum and motor unit characteristics in the masseter muscle of the rabbit. AB - Masticatory muscles contain a large variety of motor units with different physiological and morphological properties. In this study, we tested the hypothesis that a relationship exists between the mechanical and myo-electric properties of single motor units in the masseter muscle of the rabbit. It was expected that faster-contracting motor units, which usually have a relatively large number of fibers with large diameters, should have faster action potentials with larger amplitudes than slower motor units. Single motor units were stimulated. A two-dimensional force transducer registered mechanical parameters of the units. EMG electrodes were used to determine amplitude and frequency parameters of the action potentials of the same units. The results showed that faster-contracting motor units indeed produced action potentials with higher conduction velocities. However, faster motor units had no significant larger amplitude of the action potential. Small but significant positive correlations were found between the tetanic peak force and the amplitude of the action potentials. Little difference was found among the various frequency and amplitude parameters, respectively, making them equally suitable to describe the action potential. Surprisingly, a negative correlation between the amplitude and frequency parameters of the action potential was found, which may result from variability in arrival times of action potentials at the electrode site. Regional differences in the frequency parameters were found between the anterior and posterior parts of the superficial masseter. PMID- 10831098 TI - The three-dimensional cancellous bone architecture of the human mandibular condyle. AB - In the present study, we tested the hypothesis that the cancellous bone of the mandibular condyle is inhomogeneous and anisotropic. For this purpose, 11 mandibular condyles from embalmed human cadavers were scanned in a micro-CT system. Within each condyle, 9 volumes of interest were selected from different mediolateral and supero-inferior regions. Several bone parameters were calculated to describe the morphology. It appeared that the cancellous bone of the condyle could be approximated by parallel plates. These plates were almost vertically oriented at an angle of 17 degrees relative to the sagittal plane, i.e., perpendicular to the condylar axis. In the superior regions of the condyle, the cancellous bone had the largest bone volume fraction (0.19), associated with the thickest trabeculae (0.11 mm), and the highest trabecular number (1.72 mm(-1)). The lowest bone volume fraction (0.15) was found more inferiorly. The degree of anisotropy increased from superior to inferior across the condyle. No mediolateral differences in bone morphology were found, but superiorly central regions contained more bone than peripheral regions. The plate-like trabeculae could indicate that the condyle is optimally adapted to sustain loads from all directions in a plane perpendicular to the condylar axis. The high bone mass and lower anisotropy in the superior regions could enable the condyle to sustain multiple load directions. Toward the collum, the trabeculae are more aligned. This could point to stresses acting predominantly in one direction. PMID- 10831099 TI - The relationship between the density of the alveolar processes and that of post cranial bone. AB - Skeletal mass declines in all populations with age, and the literature suggests that changes in oral bone may be linked to the status of the post-cranial (systemic) skeleton. However, there is a lack of information defining the relationship between alveolar process bone and the post-cranial skeleton in healthy individuals. The purpose of this study was to determine: (1) if the bone densities of the maxillary and mandibular alveolar processes are related to the bone density of the spine, hip, or radius in healthy women; and (2) if the alveolar process densities decline with age. Forty-one dentate Caucasian women aged 20 to 78 years underwent assessment of post-cranial (systemic) and alveolar process bone. D-speed vertical bitewing and periapical radiographs incorporating aluminum stepwedges, controlled exposure and processing conditions, and a density correction algorithm were used to make alveolar process density assessments with regions of interest (ROIs) apical to crestal bone and intrabony defects. Anteroposterior lumbar (L1 to L4) and lateral lumbar (L2 to L4) spine, total hip (and subregions), and radius bone densities were determined by dual-energy x-ray absorptiometry (DEXA). Correlation analysis revealed significant relationships between maxillary alveolar process bone density and the density of the mandibular alveolar process (r = 0.57, p < or = 0.001), anteroposterior lumbar spine (r = 0.53, p < or = 0.001), lateral lumbar spine (r = 0.52, p < or = 0.001), total hip (r = 0.39, p = 0.01), total radius (r = 0.39, p = 0.01), and age (r = -0.38, p = 0.01). A two-tailed t test comparison revealed significantly greater maxillary alveolar process bone density in women younger than 50 years of age than in those 50 and older (p < or = 0.01). We conclude that the density of maxillary alveolar process bone is significantly related to the density of the mandibular alveolar process, lumbar spine, hip, and radius in healthy women and that maxillary alveolar process bone density declines with age. PMID- 10831100 TI - Self-perceived oral health status, psychological well-being, and life satisfaction in an older adult population. AB - Numerous studies have demonstrated that many older adults have problems chewing, pain, difficulties in eating, and problems in social relationships because of oral disorders. However, it is not clear if these functional and psychosocial outcomes affect broader psychological well-being and life satisfaction. Consequently, this paper begins to address the question, 'Does poor oral health compromise the quality of life?'. Initial cross-sectional analyses used data derived from the seven-year follow-up of the Ontario Study of the Oral Health of Older Adults. As at baseline and three-year follow-up, oral health was measured by self-ratings of oral health and five oral health indices. Psychological well being and life satisfaction were assessed according to the Morale Index, the Perceived Life Stress Questionnaire, The Life Satisfaction Scale, and the General Health Questionnaire. All oral health variables were significantly associated with scores from the first three of these measures in the expected direction. These associations remained after we controlled for other potential influences on the quality of life. In addition, prospective analysis indicated that self perceived oral health at three years had a significant independent effect on psychological well-being and life satisfaction at seven years. These results suggest that poor self-perceived oral health and relatively poor quality of life co-exist in the same subgroup of older adults. PMID- 10831101 TI - The association of basic proline-rich peptides from human parotid gland secretions with caries experience. AB - To address whether there are associations between the peptide composition of human parotid saliva and dental decay (caries) experience, we have characterized the peptides from parotid ductal saliva collected from nine adults who have remained free from dental caries (mean age = 59.2; Decayed Missing Filled Surfaces index [DMFS] = 0) and nine individuals who have experienced caries (mean age = 51.2; mean DMFS = 38.4). Ethanol-soluble peptides were size-fractionated on columns of Bio-Gel P-2; the salivary peptides derived from caries-susceptible subjects appeared larger than those found in the saliva of caries-free subjects. Peptides were then resolved into 19 species by cation exchange HPLC. Sequence analysis identified 18 peptides that appear to be proteolytic cleavage products of the basic proline-rich proteins IB-4, IB-5, IB-7, IB-8b, and P-B. The peptides that were more abundant in saliva obtained from the caries-free group differed from those isolated from the caries-susceptible group. The median peptide concentration of one possible precursor protein, IB-7, was found to be higher in saliva collected from caries-free individuals than in that from caries susceptible individuals. Although differences were found in the phenotypes of proline-rich proteins expressed by these groups of caries-free and caries susceptible subjects, no statistically significant associations were observed among proline-rich phenotypes and the level of any peptide. Collectively, our results indicate that proteolytic processing of parotid salivary proteins differs among individuals who have remained caries-free and those who have experienced dental decay. PMID- 10831102 TI - Effect in vitro acidification on plaque fluid composition with and without a NaF or a controlled-release fluoride rinse. AB - Plaque fluid ion concentration changes, especially fluoride, in response to the pH decrease associated with a cariogenic episode are important components of the caries process. A "controlled-release" (CR) fluoride rinse, based on the controlled release of fluoride in the presence of calcium, has been shown to form large fluoride reservoirs in resting plaque. In this study, the in vitro acid induced release of fluoride, and other ions, was examined in 48-hour-fasted plaque fluid from subjects (n = 11) who received no rinse, or who used a 228-ppm CR or NaF fluoride rinse 1 hr before being sampled. After collection, the plaque was centrifuged to yield plaque fluid, acidified (0.1 microL of 0.5 mol/L HCl per milligram plaque), and then re-centrifuged before a second sample was obtained. Although previous studies indicated a higher plaque fluid fluoride after the new rinse relative to NaF, no statistically significant difference was observed here. Average fluoride release after acidification (average pH, 5.2) was statistically greater following the use of the CR rinse (153 micromol/L) compared with the NaF rinse (17 micromol/L). No fluoride release was seen in the no-rinse samples. The pH, free calcium, phosphate, acetate, propionate, and buffer capacity were not affected by the different amounts of fluoride deposited in the plaque. However, following acid addition, an increase in free calcium and phosphate was observed, which was also independent of the rinse. The large release of fluoride following acidification suggests that the new rinse may provide an improved cariostatic effect. PMID- 10831103 TI - Remineralization effects of a two-solution fluoride mouthrinse: an in situ study. AB - Results from previous studies show that a two-solution fluoride (F) rinse is significantly more effective than a NaF rinse of the same F content of 250 microg/g (ppm) in remineralizing enamel and root lesions in an in vitro cyclic de and remineralization model. In the present study, the two-solution rinse and two NaF rinses with F contents of 250 ppm and 1000 ppm were evaluated in an intra oral remineralization model. Caries-like lesions were formed in the enamel of extracted human molars with the use of a pH 4 demineralizing solution. Thin sections of the enamel (approximately 120 microm) containing lesions were prepared, and the mineral contents of the lesions were assessed by quantitative microradiography. With the cut surfaces protected by nail varnish, 3 enamel specimens were mounted with wax in the lingual areas of a removable mandibular appliance. The study used a randomized, crossover design with seven subjects. In each of the 3 legs of the study, subjects wore the appliances continuously except when eating, drinking, and brushing their teeth. Twice daily (after breakfast and before bedtime), the subjects received a one-minute rinse with 20 mL of (1) 250 ppm-F NaF rinse, (2) 1000-ppm-F NaF rinse, or (3) 228-ppm-F two-solution F rinse. At the end of the 14-day experimental period, the sections were retrieved, and the mineral contents of the lesions were again assessed quantitatively. The results show that both the 1000-ppm-F NaF and 228-ppm-F two-solution rinses produced a greater (p < 0.05) remineralization than did the 250-ppm-F NaF rinse. The remineralization produced by the two-solution rinse was not statistically different (p > 0.05) from that produced by the NaF rinse with 4x the F content (1000 ppm F). PMID- 10831104 TI - Latency of the auditory evoked neuromagnetic field components: stimulus dependence and insights toward perception. AB - This review will focus on investigations of the auditory evoked neuromagnetic field component, the M100, detectable in the magnetoencephalogram recorded during presentation of auditory stimuli, approximately 100 milliseconds after stimulus onset. In particular, the dependence of M100 latency on attributes of the stimulus, such as intensity, pitch and timbre will be discussed, along with evidence relating M100 latency observations to perceptual features of the stimuli. Comparison with investigation of the analogous electrical potential component, the N1, will be made. Parametric development of stimuli from pure tones through complex tones to speech elements will be made, allowing the influence of spectral pitch, virtual pitch and perceptual categorization to be delineated and suggesting implications for the role of such latency observations in the study of speech processing. The final section will deal with potential clinical applications offered by M100 latency measurements, as objective indices of normal and abnormal cortical processing. PMID- 10831105 TI - Magnetoencephalographic studies of functional organization and plasticity of the human auditory cortex. AB - Magnetoencephalography has proven to be a powerful noninvasive tool for investigating the functional organization of the human auditory cortex and its plastic changes. The first part of this review summarizes some recent experiments on the tonotopic organization, which can be observed not only in the slow auditory evoked fields, but also in the middle-latency and the steady-state fields. In the second part of this review, recent studies on plasticity of the auditory cortex are outlined. These studies showed that the cortical representation of tones may change within hours after a reversible "functional deafferentation" (short-term plasticity) and that early musical training leads to an expansion in the cortical representation of complex harmonic sounds (long-term plasticity). PMID- 10831106 TI - Insights into brain function and neural plasticity using magnetic source imaging. AB - This review outlines the rationale for the use of magnetoencephalography (MEG) or magnetic source imaging (MSI), a noninvasive functional imaging technique, and the features that any imaging method should display to make a substantial contribution to cognitive neuroscience. After a brief discussion of the basic experimental approach used in the authors' studies, the use of early sensory components of brain magnetic responses is reviewed to address issues of the functional organization of the primary sensory cortices, followed by a comment on the clinical use of these components. Second, normative studies focusing on the late components of magnetic responses for establishing the validity and reliability of MSI maps of the language-specific cortex in normal subjects are reviewed. Third, the authors' investigations of fine spatiotemporal features of brain activation maps, specific to receptive language and to reading, are reviewed. Fourth, experience with presurgical mapping of the language-specific cortex in neurosurgery candidates and in patients undergoing the "Wada" procedure is summarized followed by a comment on the perfect agreement of the MSI maps with those derived by more direct invasive brain mapping procedures. Fifth. MSI derived evidence of often dramatic, functional reorganization of brain areas subserving both simple sensory and linguistic functions is summarized along with comments on the use of MSI as a means for investigating brain plasticity. Finally, in the sixth section of this review, the authors relate their experience with the use of MSI in deriving brain activation profiles during silent reading of real words and pseudowords that are specific to dyslexic children. The review concludes with a discussion on the further use of MSI in assessing, among other issues, the effectiveness of intervention strategies designed to improve reading fluency in dyslexic children. PMID- 10831107 TI - Neurophysiology of fluent and impaired reading: a magnetoencephalographic approach. AB - This article reviews a series of magnetoencephalographic (MEG) experiments aimed at identifying cortical areas and time windows relevant or even critical for fluent reading. The approach was to compare single-word processing in fluent and dyslexic readers. The activations which differed between the two groups were then studied in more detail to determine their functional roles. In fluent reading, overall visual feature processing occurs about 100 milliseconds (ms) after seeing a word, in the posteromedial extrastriate cortex bilaterally. This activation does not differentiate between letters and symbols. The first reading-specific signal is detected about 150 ms after word onset, when the left inferior occipitotemporal cortex responds preferentially to letter strings. After 200 ms, the left superior temporal cortex, in particular, is engaged in semantic processing of single words and their integration with connected text. While visual feature processing seems to be within normal limits in dyslexic subjects, reading is disrupted during the first 200 ms after seeing a word, at the letter string specific stage. The subsequent activations are weak and delayed as compared with those in fluent readers. Also presented is a case of deep dyslexia, where the same tools were used to demonstrate that reading comprehension was still subserved by the left hemisphere despite severe damage. PMID- 10831108 TI - Clinical applications of magnetoencephalography. PMID- 10831109 TI - Neuromagnetic recordings in temporal lobe epilepsy. AB - The introduction of whole-head magnetoencephalography (MEG) systems facilitating simultaneous recording from the entire brain surface has established MEG as a clinically feasible method for the evaluation of patients with temporal lobe epilepsy (TLE). In mesial TLE, two types of MEG spike dipoles could be identified: an anterior vertical and an anterior horizontal dipole. Dipole orientations can be used to attribute spike activity to temporal lobe subcompartments. Whereas the anterior vertical dipole is compatible with epileptic activity in the mediobasal temporal lobe, the anterior horizontal dipole can be explained by epileptic activity of the temporal tip cortex. In nonlesional TLE, medial and lateral vertical dipoles were found which could distinguish between medial and lateral temporal seizure onset zones as evidenced from invasive recordings. In lesional TLE, MEG could clarify the spatial relationship of the structural lesion to the irritative zone. Evaluation of patients with persistent seizures after epilepsy surgery may represent another clinical important application of MEG because magnetic fields are less influenced than electric fields by the prior operation. Simultaneous MEG and invasive EEG recordings indicate that epileptic activity restricted to mesial temporal structures cannot reliably be detected on MEG and that an extended cortical area of at least 6 to 8 cm2 involving also the basal temporal lobe is necessary to produce a reproducible MEG signal. In lateral neocortical TLE MEG seems to be more sensitive than scalp-EEG which further underlines the potential role of MEG for the study of nonlesional TLE. Whole-head MEG therefore can be regarded as a valuable and clinically relevant noninvasive method for the evaluation of patients with TLE. PMID- 10831110 TI - Magnetoencephalography in extratemporal epilepsy. AB - Epilepsy surgery candidates with extratemporal foci represent a particular diagnostic and therapeutic challenge, because of anatomic and functional features of the pertaining areas. In the last decade, novel developments in the field of electrophysiological techniques have offered new approaches to detailed localization of specific epileptic discharges as well as eloquent regions. Magnetoencephalography, in combination with neuroimaging data and simultaneously recorded EEG, yields promising results to clarify centers of epileptic activity and their relationship to structural abnormalites and functionally significant areas. Examples are given to illustrate the range of applications of this method as a contribution to routine presurgical evaluation. PMID- 10831111 TI - Somatosensory, auditory, and visual evoked magnetic fields in patients with brain diseases. AB - The features of somatosensory (SEFs), auditory (AEFs), and visual evoked fields (VEFs) in healthy subjects and patients with brain diseases provide the basis for clinical investigations using magnetoencephalography (MEG). The SEFs provide clinically useful information to identify the central sulcus and somatotopic organization of the primary somatosensory cortex. Localization accuracy of the SEFs can be tested by cortical stimulation during surgery. Functional reorganization suggested by SEF studies must be verified by other modalities. The AEFs can localize the auditory cortex in the bilateral temporal lobes. Separation of bilateral activities is much clearer in AEFs than in auditory evoked potentials. Modulation of the interhemispheric differences of latency, amplitude, and source localization of AEFs can be used to evaluate auditory function in patients with intracranial lesions. Pattern reversal VEFs provide stable localization of the primary visual function. Separation of bihemispherical activities is the advantage of VEFs over visual evoked potentials. Investigation of VEFs provides objective evaluation of visual field deficits such as homonymous or bitemporal hemianopsia in patients with intracranial lesions. Evoked magnetic fields can provide useful diagnostic information. Such clinical findings, in turn, provides the opportunity to test the source estimation accuracy of MEG. PMID- 10831112 TI - Magnetic source imaging in fixation-off sensitivity: relationship with alpha rhythm. AB - A patient in whom a variety of abnormal EEG findings can be elicited by elimination of central vision and fixation demonstrates fixation-off sensitivity. The underlying mechanisms of fixation-off sensitivity and its relationship with alpha rhythm remain unclear. To obtain a better understanding of this issue, we used a whole-head magnetoencephalograph to study an epileptic child with fixation off sensitivity resulting in a 3-Hz, large-amplitude oscillation (300 microV) over the occipital regions on the EEG. Magnetic source localization revealed alpha activity around the calcarine fissure and surrounding parieto-occipital areas. Magnetic sources of abnormalities relating to fixation-off sensitivity, however, usually were located deeper in the brain, suggesting more extensively distributed sources, with involvement of the cingulate gyrus and the basomesial occipitotemporal region. Distributions of the sources of both types of activities show independent clusters but also an appreciable domain of overlap. Our findings indicate that abnormalities related to fixation-off sensitivity can emerge in thalamocortical networks, with larger and more anterior cortical distribution than those that generate alpha rhythm. Transition in the type of oscillation appears not only to depend on a change in cellular dynamics but also to be reflected in a different spatial distribution of the underlying neuronal networks. PMID- 10831114 TI - The last word. PMID- 10831113 TI - Magnetoencephalographic characterization of sleep spindles in humans. AB - Sleep spindles in EEG recordings of adults are most prominent over the central and frontal midline regions. Early magnetoencephalographic recordings agreed with conventional EEG findings. However, more recent small-array magnetoencephalography and quantitative EEG studies suggest that the source areas for spindles are more widespread. We used a whole-head 122-channel biomagnetometer to characterize the sources of sleep spindles in four normal volunteers. Parallel interactive and automated multiple dipole spatiotemporal source modeling was conducted on the data sets of 10 spindles from each subject. Principal component analysis was used to estimate the number of sources in interactive source modeling, and singular value decomposition was used in automated dipole modeling. Spectral analysis of the epochs containing sleep spindles was performed. Principal component analysis and singular value decomposition suggested that all sleep spindles were made up of activity from multiple sources. Similarly, interactive and automated multiple dipole source modeling showed that three or more sources were present in 75% of spindle bursts. The sources for sleep spindles localized to all four cerebral lobes. Parietal and frontal lobes were the areas most frequently involved. Interactive source modeling resulted in more frequent temporal lobe than occipital dipole localizations; automated source modeling showed more frequent occipital than temporal sources. Spindle source localizations varied across subjects and across different spindles within subjects. Our results indicate that individual sleep spindles are generated by multiple cortical sources that are widespread within and across individuals. PMID- 10831115 TI - Apoptosis in rodent peri-implantation embryos: differential susceptibility of inner cell mass and trophectoderm cell lineages--a review. AB - Inner cell mass (ICM) and trophectoderm cell lineages diverge early in cleavage in response to a complex combination of cellular and molecular determinative events. The resulting differences in metabolic requirements, cell positioning and micro-environments are considered as some of the major causes underlying the differential sensitivity of ICM and trophectoderm cell lines to embryotoxic agents. In most instances, ICM cells appear less resistant to disruption than trophectoderm cells, and past observations suggest that over-stimulation of apoptosis is probably one of the mechanisms leading to selective ICM depletion at the blastocyst stage. Disproportionate deficiency in this lineage below a certain threshold level may then prevent the ICM core from providing sufficient prefetal stem cells during gastrulation and from sending regulatory signals to the trophectoderm, leading to compromised post-implantation development. The aim of this review article is to discuss the above observations and to show the value of the impact of hyperglycaemia on blastocyst metabolism and development as an exciting model for further studies. PMID- 10831116 TI - Angiogenesis in embryonic development--a review. AB - Recent studies, particularly of genetically altered mice, have greatly improved our understanding of the molecular basis underlying the development of the vascular system. Endothelial signalling systems, such as the vascular endothelial growth factor (VEGF)/VEGF receptor system and the angiopoietin/Tie2 system, were identified as central regulators of embryonic angiogenesis. The complex interactions between these and other endothelial signalling systems are beginning to emerge. Placenta (2000), 21, Supplement A, Trophoblast Research, 14, S11-S15. PMID- 10831117 TI - Regulation of placental vascular endothelial growth factor (VEGF) and placenta growth factor (PIGF) and soluble Flt-1 by oxygen--a review. AB - Morphological studies show poor placental vascular development and an increase in the mitotic index of cytotrophoblast cells in intrauterine growth restriction (IUGR). We hypothesized that the reported relatively high oxygen level in the intervillous space in contact with IUGR placental villi will limit angiogenesis by changes in vascular endothelial growth factor (VEGF) and placenta growth factor (PIGF) expression and function. Western immunoblot analysis demonstrates a diametric expression of PIGF and VEGF proteins throughout pregnancy, with P1GF levels increasing and VEGF levels decreasing, consistent with placental oxygenation. PIGF mRNA and protein is increased in IUGR as compared to gestationally matched normal placentae. Increasing oxygen tension upregulates P1GF protein in term placental villous explants, whereas hypoxia downregulates P1GF and VEGFR-1 (Flt-1) autophosphorylation in term trophoblast choriocarcinoma cell line (BeWo). Levels of soluble Flt-1 (sFlt-1) protein in supernatant of term villous explants were upregulated by 1 per cent hypoxia, whereas hyperoxia (40 per cent) decreased sFlt-1 levels, indicating that under conditions of increasing oxygen tension, PlGF function may remain unopposed. The addition of PlGF-1 to a spontaneously transformed first trimester cytotrophoblast cell line (ED27) stimulated cell proliferation while PlGF-2 had little effect. In contrast, the addition of PlGF-1 had little effect on endothelial cell proliferation while this was inhibited by PIGF-2. Taken together these changes provide a molecular explanation for the observed poor angiogenesis in the pathogenesis of IUGR. PMID- 10831118 TI - Oxygen and placental development during the first trimester: implications for the pathophysiology of pre-eclampsia. AB - During early pregnancy, placentation occurs in a relatively hypoxic environment which is essential for appropriate embryonic development. Intervillous blood flow increases at around 10-12 weeks of gestation and results in exposure of the trophoblast to increased oxygen tension (PO2). Prior to this time, low oxygen appears to prevent trophoblast differentiation towards an invasive phenotype. In other mammalian systems, oxygen tension effects are mediated by hypoxia inducible factor-1 (HIF-1). We found that the ontogeny of HIF-1alpha subunit expression during the first trimester of gestation parallels that of transforming growth factor-beta3 (TGFbeta3), an inhibitor of early trophoblast differentiation. Expression of both molecules is high in early pregnancy and falls at around 10 weeks of gestation when placental PO2 levels are believed to increase. Antisense induced inhibition of HIF-1alpha inhibited the expression of TGFbeta3, and stimulated extravillous trophoblast (EVT) outgrowth and invasion. Of clinical significance we found that TGFbeta3 expression was increased in pre-eclamptic placentae when compared to age-matched controls. Significantly, inhibition of TGFbeta3 by antisense oligonucleotides or antibodies restored the invasive capability to the trophoblast cells in pre-eclamptic explants. We speculate that if oxygen tension fails to increase, or trophoblasts do not detect this increase, HIF-1alpha and TGFbeta3 expression remain high, resulting in shallow trophoblast invasion and predisposing the pregnancy to pre-eclampsia. Effective fetal maternal interactions during early placentation are critical for a successful pregnancy. Optimal placental perfusion requires the controlled invasion of trophoblast cells deep into the decidua to the spiral arteries. Trophoblast stem cells, also referred to as cytotrophoblast cells, reside in chorionic villi of two types, floating and anchoring villi. Floating villi, which represent the vast majority of chorionic villi, are bathed in maternal blood and primarily perform gas and nutrient exchange for the developing embryo. During early placentation, cytotrophoblast cells in the floating villi proliferate and differentiate by fusing to form the multinucleate syncytiotrophoblast layer. Cytotrophoblast cells in anchoring villi either fuse to form the syncytiotrophoblast layer, or break through the syncytium at selected sites and form multilayered columns of non polarized extravillous trophoblast cells, which physically connect the embryo to the uterine wall (Figure 1). The extravillous trophoblast cells invade into the uterine wall as far as the first third of the myometrium and its associated spiral arteries, where they disrupt the endothelium and the smooth muscle layer and replace the vascular wall. This results in the conversion of the narrow calibre arteries into distended uteroplacental arteries, thereby increasing blood flow to the placenta and allowing an adequate supply of oxygen and nutrients to the growing fetus. The invasive activity of the extravillous trophoblast cells is at a maximum during the first trimester of gestation, peaking at around 10-12 weeks and declining thereafter. Insufficient invasion contributes to the development of pre-eclampsia, which often results in fetal intrauterine growth restriction, maternal hypertension and proteinuria. In contrast, unrestricted invasion is associated with premalignant conditions, such as invasive mole, and with malignant choriocarcinoma. Invading trophoblast cells undergo striking and rapid changes in cellular functions that are temporally and spatially regulated along the invasive pathway (Figure 1) (Cross, Werb and Fisher, 1994. The formation of the anchoring villi is accompanied by changes in synthesis and degradation of extracellular matrix proteins and their receptors, and changes in the spatial distribution of extracellular matrix proteins, as well as changes in the expression of adhesion molecules (Damsky, Fitzgerald and PMID- 10831119 TI - Placental oxygen consumption. Part I: in vivo studies--a review. AB - At term of pregnancy, oxygen consumption by the human or ovine placenta accounts for 40 per cent of total oxygen uptake by the gravid uterus. In the sheep, most oxygen is used for oxidative phosphorylation of glucose; the remainder is probably utilized for non-mitochondrial processes. The ATP yield is expended mainly in protein synthesis and cation transport. The fractional protein synthesis rate of ovine placenta is 60 per cent per day. Applying these data to man, protein synthesis is estimated to account for about 30 per cent of placental oxygen uptake. Probably this reflects the high rates of synthesis of peptide and steroid hormones. The Na+ gradient is the basis for secondary active transport of amino acids and other substances, and the Na(+)-K(+)-pump probably accounts for 20-30 per cent of oxygen uptake, with a smaller contribution from Ca(2+)-ATPase. Placental oxygen uptake remains constant during acute reductions in uterine oxygen supply and is maintained at the expense of the fetus. In the longer term, in experimental models of fetal growth restriction, placental oxygen consumption is reduced to a greater extent than fetal oxygen consumption. Placental oxygen consumption is greatly reduced under in vitro experimental conditions, due largely to an inadequate oxygen supply. This results in reduced protein synthesis and possibly inhibition of Na(+)-K(+)-ATPase. However, if the placenta is subjected to hyperoxia, by raising the PO2 of the medium, there is an increase in anaerobic glycolysis and structural damage may ensue. Premature exposure of trophoblast to high oxygen tensions in vivo may result in reduced villous branching, but this is likely to be a cause, rather than a consequence, of reduced fetal growth and oxygen consumption. PMID- 10831120 TI - Placental oxygen consumption. Part II: in vitro studies--a review. AB - The human placenta in vivo consumes large amounts of oxygen. After delivery the placenta is exposed to anoxia and under in vitro experimental conditions oxygen consumption is only a fraction of in vivo estimates. In spite of a reduced oxygen supply, structural and functional integrity of the tissue is surprisingly well preserved. Special metabolic adjustments in the sense of 'partial metabolic arrest' may be the explanation for a remarkable survival capacity of placental tissue and reduction of protein synthesis seems to be an important component of metabolic slowdown. The potential significance of this special feature of placental metabolism for the in vivo situation is discussed. PMID- 10831121 TI - Post-implantation differentiation and proliferation of cytotrophoblast cells: in vitro models--a review. AB - Cytotrophoblast cells, specialized placental cells, proliferate early in pregnancy and then differentiate into tumour-like cells that invade the uterus and its vasculature. We have established in vitro models of three-dimensional cultures for anchoring villi and cell islands on extracellular matrix in order to study regulation of cytotrophoblast cell differentiation and proliferation. It has been demonstrated that cytotrophoblast cells from cell islands and cell columns share the same characteristics and that their differentiation is triggered by interaction with the extracellular matrix. The fact that during much of the first trimester maternal blood flow to the placenta is at a minimum, suggests that oxygen tension might regulate cytotrophoblast proliferation and differentiation. Hypoxia, comparable to that encountered by early gestation cytotrophoblast cells in the intervillous space, stimulated the cells to enter the cell cycle and inhibited their differentiation along the invasive pathway. Thus, oxygen gradient and cell-matrix interactions at the maternal-fetal interface play an important role in the regulation of cytotrophoblast proliferation and differentiation. PMID- 10831122 TI - The homeobox genes MSX2 and MOX2 are candidates for regulating epithelial mesenchymal cell interactions in the human placenta. AB - Homeobox genes of the Msx and Mox families are coexpressed in the vertebrate embryo in regions of epithelial-mesenchymal interactions. Here we show that a member of each family is expressed in extra-embryonic structures where epithelial and mesenchymal cell layers contact. In situ hybridization studies on first trimester human placental sections reveal that MSX2 and MOX2 are expressed predominantly in the cytotrophoblast cell layer. In term placenta, MSX2 and MOX2 are expressed in the syncytiotrophoblast. This is the first study to describe the expression of MOX2 in human tissues and to show that members of the Msx and Mox families of homeobox genes are expressed where epithelial and mesenchymal cell layers contact in the human placenta. A combinatorial code of homeobox genes that includes members of the Msx, Mox and Dlx families has been predicted to regulate epithelial-mesenchymal cell interactions in the vertebrate embryo. We have shown that MSX2, MOX2, DLX4 and the HB24 homeobox gene are expressed in the epithelial and mesenchymal cell types that form the placenta. We predict that this combination of homeobox genes is involved in regulating epithelial-mesenchymal cell interactions in extraembryonic tissues. PMID- 10831123 TI - Paracrine and autocrine regulators of trophoblast invasion--a review. AB - Cytotrophoblastic cells (CTBs) from first trimester placenta follow one of two existing differentiation pathways: villous CTBs (vCTBs) form a monolayer of polarized epithelial stem cells which proliferate and eventually differentiate by fusion to form a syncytiotrophoblast (STB) covering the entire surface of the villus, or they can break through the STB at selected sites (in anchoring villi) to form multilayered columns of non-polarized but invasive CTBs. In vitro, CTBs invade a reconstituted basement membrane, they thus behave like metastatic cells. This invasive behaviour is due to the ability of CTBs to secrete matrix metalloproteinases (MMPs) since tissue inhibitor of MMP (TIMP) inhibits their invasiveness. MMPs are a family of at least 17 human zinc-dependent endopeptidases collectively capable of degrading essentially all components of the extracellular matrix (ECM). Although CTBs behave like metastatic cells, in vivo they are only transiently invasive (first trimester) and their invasion is normally limited only to the endometrium and to the proximal third of the myometrium. This temporal and spatial regulation of trophoblast invasion is believed to be mediated in an autocrine way by trophoblastic factors and in a paracrine way by uterine factors. Several types of regulators have been investigated: hormones, cytokines, growth factors and ECM glycoproteins. This review is not intended to be an exhaustive catalogue of all the potential regulators but is aimed at emphasizing those factors relevant in trophoblast endometrial interactions. PMID- 10831124 TI - Cell-cell-communication during placental development and possible implications for trophoblast proliferation and differentiation. AB - Since direct cell-cell-communication plays a crucial role in the coordination of proliferation and differentiation processes during development we have focused on the expression patterns of gap junctions and their functional implication in the human placenta. The gap junction protein connexin40 (Cx40) is expressed in the proximal extravillous trophoblast of cell islands and columns. In accordance with these observations, isolated trophoblast cells from first and second trimester placentae and choriocarcinoma cells (Jeg-3) reveal Cx40 expression. This channel is not only characteristic of the trophoblast cells along the invasive pathway but also of endothelial cells. To elucidate the functional role of this channel for proliferation and invasion, the non-coupled Jeg-3 cells have been transfected with Cx26, Cx40 and Cx43, respectively. In contrast to Cx40, the Cx26 channel was more potent in reducing proliferation and inducing differentiation indicated by hCG-beta secretion. Using the nude mouse model to study invasion properties of choriocarcinoma cells, we demonstrated that malignant trophoblast cells were able to invade host vessels and to replace endothelial cells. Upregulation of endogeneous connexin genes in tumours grown in nude mice enforces further experimental strategies to investigate the importance of the different channels to fake the cell biological program of endothelial cells. PMID- 10831125 TI - Angiogenesis and placental vasculature--a workshop report. PMID- 10831126 TI - Novel genes and transcription factors in placental development--a workshop report. PMID- 10831127 TI - Trophoblast apoptosis and placental development--a workshop report. PMID- 10831128 TI - Transplantation into genetically alymphoid mice as an approach to dissect the roles of uterine natural killer cells during pregnancy--a review. AB - Mice genetically deficient in the natural killer (NK) cell lineage lack uterine (uNK cells) and demonstrate morphometrically-quantifiable histopathology within their implantation sites. Two particular mouse strains, tg(epsilon),26 and RAG-2 null x gamma(c) null, have been used successfully as transplant recipients to address questions relating to the biology of uNK cells. uNK cells did not differentiate within decidualized uterine graft segments from normal mice, which were anastomosed orthotopically into immunodeficient hosts. uNK cells did appear in similar grafts placed into immunocompetent hosts, indicating that uNK cells or their progenitors must home to the uterus. This was confirmed by splenocyte transplantation into pregnant uNK cell deficient recipients. Only splenocytes from pregnant donors, not those from non-pregnant donors, homed to the uterus. Homing in this in vivo assay was independent of the CC-chemokine receptors, CCR-2 and CCR-5. Longer-term bone marrow cell reconstitution of neonatal or virgin adult uNK cell-deficient mice has identified a functional role for uNK cells in modification of the decidual arterioles which is mediated by IFN-gamma. By utilizing mutant and gene-ablated mice as donors for tissue or haematopoietic cell transplants to uNK cell deficient mice, it should be possible to fully characterize the in vivo regulation and functions of these pregnancy-specific uterine lymphocytes. PMID- 10831129 TI - Recognition of trophoblast HLA class I molecules by decidual NK cell receptors--a review. AB - During placentation the extravillous trophoblast (EVT) cells migrate through the decidua towards the maternal spiral arteries. The walls of the arteries are then destroyed by trophoblast resulting in an increased blood flow to the fetus. These EVT express HLA-G, HLA-E and HLA-C, an unusual combination of two non-classical and one classical MHC class I molecules. The decidua is infiltrated by distinctive uterine natural killer (NK) cells during the time of trophoblast invasion. These cells express a variety of receptors (CD94/NKG2, KIR and ILT) which are known to recognize HLA class I molecules. There is, therefore, a mechanism for molecular recognition of the placental trophoblast cells. The possible functional consequences of this uterine NK cell-trophoblast interactions are uncertain. One possible result is in an altered NK cell cytokine profile which modulates the invasive proclivity of the EVT. In this way placentation could be controlled. PMID- 10831130 TI - HLA-G polymorphisms and molecule function--questions and more questions--a review. AB - Human leukocyte antigen (HLA)-G is a non-classical HLA-class I antigen which is predominantly expressed on invasive trophoblastic cells and is postulated to be a mediator of maternal-fetal tolerance. HLA-G interacts with NK cells, can present nonamer peptides and binds CD8 in an analogous manner to classical HLA-I. The HLA G protein exists in soluble and membrane-bound isoforms generated through alternative splicing. Although initially considered to be non-polymorphic, variations of the HLA-G DNA sequence have been reported which led to the definition of a limited number of HLA-G alleles including the Null-allele G*0105N. Whereas the HLA-G DNA sequence shows a high degree of conservation in positions which are essential for classical HLA-I molecule functions, polymorphic sites in HLA-G are not congruent with sites of high nucleotide variability in classical HLA. The identification of two females with recurrent spontaneous abortions who are homozygous for the G*0105N Null-allele re-opens the discussion about the role of HLA-G in pregnancy and underlines the need of a systematic analysis of the different hypotheses of HLA-G function in vivo. PMID- 10831131 TI - HLA class I molecules in the placenta: which ones, where and what for? A workshop report. PMID- 10831132 TI - Roles of cytokines and immune cells at the interface--a workshop report. PMID- 10831133 TI - DNA microarrays: a novel approach to investigate genomics in trophoblast invasion -a review. AB - The events that regulate trophoblast invasion need to be characterized at the transcriptional level. Several types of gene products may be involved in various stages oftrophoblast infiltration, including integrins, matrix metalloproteases (MMPs) and extracellular matrix (ECM) proteins. Autocrine or paracrine regulators of cytotrophoblast proliferation or differentiation in vitro (e.g. growth factors and cytokines, as well as oxygen tension) could be characterized mechanistically at the transcriptional level. Large-scale gene expression profiling of trophoblasts of distinct invasive stages could be carried out on fixed tissue obtained by laser-directed microdissection. This information may shed light on physiological implantation and placentation, as well as on the interpretation of pathological processes such as pre-eclampsia. The applications of DNA microarrays are ideal for studies of genomic structure (e.g. mutation and polymorphism analyses) and monitoring of gene expression. The ultimate goal is to understand the critical events underlying growth, development, homeostasis, 'behaviour and the onset of disease at a genomic level. Microarrays detect gene expression levels in parallel by measuring the hybridization of labelled, single-stranded DNA to many thousands of partial or whole gene sequences immobilized on a glass surface (the 'chip'). Microarrays are available both commercially and can be manufactured in house. PMID- 10831134 TI - Phage display: a molecular tool for the generation of antibodies--a review. AB - Phage display comprises an array of methods, which can be used to display proteins on bacteriophages. We present in this article a summary of techniques, which can be used to express antibody libraries on bacteriophages. Since many researchers are more familiar with the conventional hybridoma technique for production of monoclonal antibodies we describe analogies and differences between these two techniques, both of which are used to reach comparable scientific objectives. We focus on the features which are specific to phage display techniques rather than for hybridoma techniques. These comprise the freedom to choose other animals than the mouse for immunization, the enormously large sample (up to 10(9) clones) which can be drawn and immortalized from a single immunized animal and the opportunity to enhance affinity of isolated antibodies by in vitro affinity maturation. The panning techniques, which are used to enrich specifically binding phage antibodies from the huge libraries are briefly summarized. PMID- 10831135 TI - Cell culture models of trophoblast II: trophoblast cell lines--a workshop report. PMID- 10831136 TI - Cell culture models of human trophoblast--primary culture of trophoblast--a workshop report. PMID- 10831137 TI - Leisure activities and adolescent antisocial behavior: the role of structure and social context. AB - The goal of this study was to understand better how the structure and social context of adolescent leisure activities relates to antisocial behavior. A representative sample of 703 14-year-olds and their parents were assessed concerning adolescent involvement in community-based leisure activities, peer and adult social relations, and antisocial behavior. Results showed that participation in highly structured leisure activities was linked to low levels of antisocial behavior, while participation in activities with low structure (i.e. a youth recreation center) was associated with high levels of antisocial behavior. Overall the results were similar for boys and girls; however, the combination of involvement in a low structured activity and the absence of any highly structured participation appeared especially problematic for boys' antisocial behavior. Participants of low structured activities were also characterized by deviant peer relations, poor parent-child relations, and they received low support from their activity leader compared to adolescents engaged in more structured community activities. Findings are discussed in terms of their implication for prevention research. PMID- 10831138 TI - Prevalence and correlates of smokeless tobacco use in a sample of Connecticut students. AB - We examined the prevalence of smokeless tobacco use, and its relationship to other risky behaviors in Connecticut school children in grades four through 12 (n=31,861). Less than one per cent of students in grades 4-8, and four per cent of those in grades 9-12, used smokeless tobacco. Eighty-two percent of all users were white males. Among high school males, the prevalences of risk-taking behaviors and feelings of stress and depression were significantly higher among users than non-users. Among high school males, the highest prevalences of risky behaviors and stress indicators were reported by joint users of cigarettes and smokeless tobacco. PMID- 10831139 TI - Pathological attachment and attachment style in late adolescence. AB - Attachment theory is frequently invoked to account for patterns of adaptation within relationships. West and Sheldon derived a measure of dysfunctional adult attachment from Bowlby's theory. Four patterns are identified: compulsive self reliance; caregiving; care-seeking; and angry withdrawal. The aim of this study was to assess the psychometric properties of this promising measure, and to assess its ability to predict symptomatology relative to measures of attachment style. Participants included 209 late adolescents who were involved in committed relationships. They responded to categorical and continuous measures of attachment style and various measures of symptomatology and college adjustment. The four dimensions of pathological attachment showed strong internal consistency and few gender differences. Compulsive care-seeking, angry withdrawal and compulsive self-reliance were particularly strong predictors of psychiatric symptomatology and insecure attachment style. Attachment pathology and insecure attachment style predicted symptomatology similarly, though variations were evident depending on how style was assessed. This study is the first to provide evidence of convergent, discriminant and predictive validity for this measure. Directions for future research are noted. PMID- 10831140 TI - Denial of HIV/AIDS and preventive behaviour among Israeli adolescents. AB - A national sample of 1082 Israeli adolescents, aged 14-18, was questioned on the following topics: Respondents' sexual habits and HIV/AIDS prevention behaviours, HIV/AIDS-related knowledge and attitudes, and six kinds of HIV/AIDS-related denial. The main results showed that infrequent condom use was associated with a high level of denial, denial of personal relevance and of responsibility being the most salient. These results suggest that denial may constitute a major factor in HIV/AIDS-related risk behaviours, with implications for intervention programmes aimed at adolescents. PMID- 10831141 TI - An experimental investigation of the role of appraisal processes in adolescents and their mothers experiencing family conflict. AB - A social information processing perspective was used to investigate appraisal processes in family conflict situations. Fifty-five young adolescents (32 males and 23 females) and their mothers who were experiencing sufficient conflict to seek mediation assistance, were presented with hypothetical scenarios depicting common sources of conflict. Participants were randomly assigned to two time conditions. After reading each scenario, participants chose one of four interpretations (two hostile and two non-hostile). Time condition did not influence the number of hostile interpretations for mothers, but when instructed to respond quickly, adolescents were more likely to interpret their mother's behaviour as hostile. The findings highlight the need for investigation of specific stressors which influence appraisal processes in family conflict. PMID- 10831142 TI - Personal concerns and their causes: perceptions of Hong Kong Chinese adolescent students. AB - This study investigated the personal concerns and causes of difficulties perceived by Hong Kong adolescent students. A survey of 2103 secondary school students in Year 1 to Year 3 indicated that both students' concerns and causes are multi-dimensional. Academic achievement was perceived as the most pressing concern, while problems at home and maladjusted behaviour were seen as lesser concerns. Students attributed their difficulties more to personal deficiencies, and least to family factors. Results also showed significant gender, age and school banding effects. Implications of the findings for educationalists and psychologists working with adolescents in school contexts are discussed. PMID- 10831143 TI - Parenting styles and adolescents' achievement strategies. AB - The aim of the study was to investigate the extent to which adolescents' achievement strategies are associated with the parenting styles they experience in their families. Three hundred and fifty-four 14-year-old adolescents completed a Strategy and Attribution Questionnaire and a family parenting style inventory. Analogous questionnaires were also completed by the adolescents' parents. Based on adolescents' report of the parenting styles, four types of families were identified: those with Authoritative, Authoritarian, Permissive, and Neglectful parenting styles. The results further showed that adolescents from authoritative families applied most adaptive achievement strategies characterized by low levels of failure expectations, task-irrelevant behaviour and passivity, and the use of self-enhancing attributions. Adolescents from neglectful families, in turn, applied maladaptive strategies characterized by high levels of task-irrelevant behaviour, passivity and a lack of self-enhancing attributions. The results provide a basis for understanding some of the processes by which parenting styles may influence adolescents' academic achievement and performance. PMID- 10831144 TI - Religiosity and sexual risk-taking behavior during the transition to college. AB - The degree to which religious identity acts as a protective buffer against sexual risk-taking in late adolescence was investigated in 230 first-year college students. Allport and Ross' Religious Orientation Scale was used to examine the relationship between religiosity, and sexual activity and condom use. Results indicate that greater intrinsic and extrinsic religiosity were associated with less sexual activity and condom use. Therefore religious identification may protect against initiating sexual activity among late adolescents, but may fail to protect against practicing unsafe sex among students who are already sexually active. PMID- 10831145 TI - Social influences and female adolescent dieting. AB - A large sample of female adolescents whose ages ranged from 12 to 17 years completed a battery of measures assessing their dieting status and their perceived psychosocial influences to diet. Higher levels of perceived influence from parents and peers were reported by girls who were dieting with a greater frequency. While age and social influences significantly predicted dieting status on their own, when examined together, the effect of social influence overrode that for age. PMID- 10831146 TI - Differential diagnosis of chronic airflow obstruction. AB - Chronic obstructive pulmonary disease is a syndrome including illnesses such as asthma, chronic bronchitis, and emphysema. Although these diseases share a common obstructive component, their optimal treatment and prognosis differ. This article examines the salient features of the history, physical exam, pulmonary function tests, and radiological evaluation which may allow the clinician to differentiate the various diseases that make up COPD; thus allowing the clinician to better target the multiple therapeutic modalities available. PMID- 10831147 TI - Compliance during long-term treatment with fluticasone propionate in subjects with early signs of asthma or chronic obstructive pulmonary disease (COPD): results of the Detection, Intervention, and Monitoring Program of COPD and Asthma (DIMCA) Study. AB - In a prospective study, we investigated the long-term compliance to fluticasone propionate (FP) by dry powder inhalation (Rotadisk) in subjects with early signs of asthma and chronic obstructive pulmonary disease (COPD) without an established diagnosis. Subjects were selected from a large screening program on early stages of asthma and COPD (Detection, Intervention, and Monitoring Program of COPD and Asthma [DIMCA] program) in the general practice. Forty-eight adult subjects with "early signs of COPD" (slightly increased forced expiratory volume in 1 sec [FEV1] decline of >0.04L/year) and 29 adult subjects with "early signs of asthma" (signs of bronchial hyperresponsiveness or reversibility) participated in a randomized placebo-controlled trial with FP (Flixotide 500 microg daily) versus placebo with a duration of 2 years or 1 year, respectively. Compliance was measured by counting Rotadisks returned. By means of a questionnaire, participants were asked about perceived effects and/or side effects of the trial drug. The mean overall individual compliance rates of 72% (range 7%-102%) in the early COPD trial and 71% (range 8%-99%) in the early asthma trial were maintained throughout the study. Perceived effectiveness (12% of the participants) or side effects (30% of the participants) of the trial drug were not related to compliance. The willingness of patients to use the trial drug in daily practice if efficacy would be proved was statistically significantly related to compliance during the trial (p = 0.017). It was concluded that the compliance rates found were relatively high in patients with symptoms of mild asthma or COPD without an established diagnosis. Conviction of the importance of treatment influenced compliance more positively than perceived (side) effects. These results again emphasize the importance of patient education in establishing early treatment with inhaled corticosteroids. PMID- 10831148 TI - The management of asthma: a case-scenario-based survey of family physicians and pulmonary specialists. AB - This study assessed family physicians' and pulmonary specialists' approaches to the treatment of adult outpatient asthma using a self-administered questionnaire consisting of six asthma scenarios of varying severity levels. One hundred sixty three randomly selected family physicians and pulmonary specialists completed the questionnaire (response rate of 80%). We observed that, regardless of asthma severity, more than 75% of physicians (regardless of specialty) would not include oral theophylline or nonsteroidal anti-inflammatory preparations in their treatment approach. Pulmonary specialists' and family physicians' approaches to mild asthma were similar (more than 90% recommended an inhaled beta2-agonist). However, considerable differences existed among and between physician groups for the remaining scenarios. For example, with an exacerbation associated with an upper respiratory tract infection, family physicians were more likely to recommend oral antibiotics (p<0.0001) and a same-day outpatient visit (p<0.0001), whereas specialists were more likely to increase the inhaled corticosteroid dosage (p<0.0001). Overall, disagreement was observed almost twice as often among family physicians than among specialists. Our results suggest that physicians vary markedly in their reported use of most interventions available to treat asthma, even when the disease severity is specified. PMID- 10831149 TI - Implementing continuing education strategies for family physicians to enhance asthma patients' quality of life. AB - The goal of the study was to provide asthma-related continuing medical education (CME) to family physicians with the objective of improving patient outcomes. Using a quasi-experimental design in a single community, the intervention included academic detailing, a case-based workshop, newsletters, medical grand rounds, and patient-centered education materials. Outcome measures included physician participation in CME; patient self-reported quality of life, asthma knowledge, asthma self-management, medication use, and health service utilization before and after the intervention; and physician feedback. Our results indicated that 78% of family physicians participated in one or more of the CME activities. The majority of physicians provided positive feedback on the use of the intervention both from their own and their patients' perspectives. Academic detailing increased the involvement of physicians in CME. We concluded that there was a statistically significant improvement in patients' quality of life, whereas changes in patients' knowledge, behavior, and health service use were positive but not statistically significant. Methodological factors are identified that could improve the effectiveness of future studies. PMID- 10831150 TI - Asthma mortality rates among California youths. AB - Asthma mortality rates for California youths (ages 5-17 years) were examined over the period 1981-1995. Data were analyzed by Poisson regression from the California Departments of Vital Statistics, Finance, and the U.S. Census Bureau. Aggregate statewide mortality demonstrated a 2.01%, statistically insignificant, upward trend without geographic clustering. Stratified race/ ethnicity rates differed significantly, with African-American mortality twice that of the Caucasian population. In addition, an increased number of deaths occurred in households of African-Americans, Caucasians (including Hispanics), and Hispanics with incomes below statewide medians. This study on California youths differs from reports on larger populations with its minimal aggregate increase in asthma mortality, yet mirrors national trends of disproportionate impact on African Americans and the economically disadvantaged. PMID- 10831151 TI - Irritant-induced asthma: clinical and functional aspects. AB - We report on three patients who experienced persistent asthma symptoms after repetitive irritant exposure which took place over a period from several days to months. Airway inflammation was assessed by induction of sputum and functional follow-up information was obtained from serial lung function tests. All patients had bronchial hyperresponsiveness to methacholine at the time of diagnosis. However, induced sputum samples did not show increased differential count of eosinophils. Treatment with inhaled corticosteroids was started in all of the patients and two of them were removed from work. In the two patients who left the workplace, methacholine inhalation test became negative when symptoms disappeared, whereas the patient who continued working had persistent asthma symptoms and a deterioration of bronchial hyperresponsiveness. PMID- 10831152 TI - Symptomatic vascular rings in adulthood: an uncommon mimic of asthma. AB - Symptomatic thoracic vascular rings presenting in adulthood are thought to be rare. During a 3-year time period, we diagnosed four cases of symptomatic vascular rings, which had been treated unsuccessfully for suspected asthma. Spirometry was characterized by normal forced vital capacity (FVC), forced expiratory volume in 1 sec (FEV1), and FEV1/FVC, decreased peak expiratory flow (PEF), and truncation of the expiratory flow volume loop. Chest radiographs revealed a right aortic arch in each case with computed tomography (CT) or magnetic resonance imaging (MRI) confirming the diagnosis of a vascular ring. The specific abnormalities consisted of right aortic arch with mirror branching of the main arteries and persistent ligamentum arteriosum; right aortic arch with diverticulum and a fibrous embryonic left arch; right aortic arch with aberrant left subclavian artery arising from a diverticulum of Kommerell; and a right aortic arch with persistent ligamentum arteriosum. Although they are uncommon, vascular rings first presenting in adulthood as a mimic of asthma are not rare. This diagnosis should be considered in adults when abnormal truncation of the flow-volume loop occurs or when radiographic aortic arch abnormalities are found. PMID- 10831153 TI - Inhibitory activity of fenoterol on Dermatophagoides-, Parietaria-, tetanus toxoid-, and Candida albicans-stimulated blood mononuclear cells: differences in beta2-adrenoreceptor stimulation but not in cell apoptosis. AB - beta2-adrenoreceptor agonists have the ability to downregulate in vitro the proliferative response of peripheral blood mononuclear cells (BMCs). This activity could be related to a variety of beta2-adrenoreceptor-mediated functions, including induction of cell apoptosis in activated T-cells. To test this hypothesis, BMCs from atopic subjects, sensitized to house dust mites (Dermatophagoides [Der p]) and/or to Parietaria were incubated with fenoterol (10(-8)-10(-5) M) in the presence of (a) purified allergen extracts (Der p [5 microg/mL] or Parietaria [5 microg/mL]) or (b) antigens (tetanus toxoid [1 microg/mL] or Candida albicans [5 x 10(5) bodies/mL]). The BMC proliferation was assessed by [3H] thymidine incorporation and cell apoptosis was assessed by evaluating DNA fragmentation by a fluorescence technique, using propidium iodide. In cultures stimulated with Der p or with Parietaria, fenoterol induced a dose dependent inhibition of BMC proliferation, significant also at the lowest concentration tested (10(-8) M) (p < 0.05, each comparison). In contrast, the inhibitory activity of the drug on tetanus-toxoid-stimulated BMCs was significant only at the highest dose tested (10(-5)M) (p < 0.05), whereas no effect was seen when BMCs were stimulated with C. albicans extract (p > 0.05). The different inhibitory efficacy of fenoterol appeared to be related to the degree of activation of beta2-adrenoreceptors on the different BMC populations that responded to the different stimuli. Indeed, in the presence of fenoterol (10(-6) and 10(-5)M), a significant increase in cyclic adenosine monophosphate (cAMP) levels was seen in Der p- or Parietaria-stimulated cells (p < 0.05; each comparison), but not in cell cultures stimulated with tetanus toxoid or with C. albicans extracts (p > 0.05; each comparison). Finally, the percentage of cells with fragmented DNA was lower in cultures stimulated with Der p or Parietaria than in those stimulated with tetanus toxoid or C. albicans, and the presence of fenoterol did not modify cell apoptosis (p > 0.05; each comparison). Thus, the different inhibitory activity of fenoterol on BMCs activated by allergens (Der p or Parietaria) or by antigens (tetanus toxoid or C. albicans) seems to be related to differences in beta2-adrenoreceptor expression and/or function in the different antigen-specific T-cell subsets, but it is not influenced by changes in cell apoptosis. PMID- 10831154 TI - Evidence for melatonin synthesis in mouse and human bone marrow cells. AB - Recently, it was demonstrated that inbred strains of mice have a clearcut circadian rhythm of pineal and serum melatonin. Moreover, it is known that melatonin is involved in many immunoregulatory functions. Among them, hematopoiesis is influenced by the action of melatonin via melatonin-induced opioids on kappa-opioid receptors, which are present on stromal bone marrow cells. Therefore, the present study was carried out to investigate the presence of melatonin in the bone marrow in which immunocompetent cells are generated. Specifically, we aimed at answering the following question: are bone marrow cells involved in melatonin synthesis? In the present study, we demonstrate that (1) bone marrow cells contain high concentrations of melatonin; (2) bone marrow cells have a N-acetyltransferase activity and they express the mRNA encoding hydroxy-O methyltransferase and (3) bone marrow cells cultured for a prolonged period exhibited high levels of melatonin. Results presented here suggest that mouse and human bone marrow and bone marrow cells are capable of de novo synthesis of melatonin, which may have intracellular and or paracrine functions. PMID- 10831155 TI - Demonstration of hydroxyindole-O-methyltransferase (HIOMT) mRNA expression in pineal parenchymal tumors: histochemical in situ hybridization. AB - The expression of hydroxyindole-O-methyltransferase (HIOMT), an enzyme catalyzing the final step of melatonin biosynthesis, was examined in three pineoblastomas and five pineocytomas by in situ hybridization analysis. Distinct hybridization signals for HIOMT mRNA, though weaker than in normal pineal gland pinealocytes, were detected in two of the three pineoblastoma and three of the five pineocytoma cases. Of the pineoblastomas, hybridization signals were observed in most tumor cells of one case, while in another, signals were detected in occasional cells clustered or scattered throughout the neoplastic field. Of the pineocytomas, signals were detected in most tumor cells of two cases, while in one case, signals were detected only in occasional cells. Among these specimens, one pineoblastoma and one pineocytoma were also analyzed using northern blot and reverse transcription polymerase chain reaction (RT-PCR) analyses. In the northern blot analysis, an apparently single band corresponding to the size of HIOMT mRNA was detected in both pineoblastoma and pineocytoma RNA blots. In the RT-PCR analysis, three species of HIOMT mRNA generated via alternative splicing were detected in both tumors. These results suggest that the neoplastic cells of pineoblastomas and pineocytomas often retain the ability to express HIOMT mRNA, as in normal pinealocytes, and that HIOMT is a useful tumor marker for the diagnosis of pineal parenchymal tumors. PMID- 10831156 TI - Differential responsiveness of MCF-7 human breast cancer cell line stocks to the pineal hormone, melatonin. AB - The estrogen receptor (ER)-positive MCF-7 human breast cancer cell line has been used extensively for the study of estrogen-responsive human breast cancer. However, various levels of estrogen responsiveness have been described in different stocks of MCF-7 cells. Because we have previously shown that the pineal hormone, melatonin, inhibits proliferation of MCF-7 cells and can modulate ER expression and transactivation, we investigated if various stocks of MCF-7 cells exhibit a differential responsiveness to the anti-proliferative effects of melatonin and the possible mechanisms involved. The MCF-7 stocks (M, O, H) were examined for: (1) mitogenic response to estradiol; (2) steady-state ER mRNA levels; (3) expression of the mt1 melatonin membrane receptor; (4) growth inhibition by melatonin; and (5) melatonin's modulation of expression of the ER and the estrogen-regulated genes, PgR, TGFbeta and pS2. For all of these parameters, there was a stock-specific response which showed: MCF-7M > MCF-7O > MCF-7H. These results demonstrate that there are significant differences in the responsiveness of various stocks of MCF-7 breast cancer cells to the growth inhibitory effects of melatonin which can be correlated with both the level of ER mRNA expression and the degree of estrogen-responsiveness. These findings suggest that not only may these differences have some impact on the cells' estrogen response pathway, but also that the primary growth-inhibitory effects of melatonin are transduced through the membrane-associated G-protein coupled mt1 melatonin receptor. PMID- 10831157 TI - Stimulatory and entraining effect of melatonin on tuberoinfundibular dopaminergic neuron activity and inhibition on prolactin secretion. AB - The aims of the present study were to determine if melatonin exerts an effect on prolactin (PRL) secretion via the tuberoinfundibular dopaminergic (TIDA) neurons and if endogenous or exogenous melatonin has an entraining effect on the rhythmic changes of TIDA neuronal activity and PRL secretion. Melatonin given in the morning (10:00 h), dose- (0.01-1 mg/kg, ip) and time- (at 15 and 60 min, but not at 30 min) dependently stimulated TIDA neuronal activity in ovariectomized (OVX), estrogen-treated rats as determined by 3,4-dihydroxyphenylacetic acid (DOPAC) levels in the median eminence (ME). Serum PRL was concurrently inhibited by the injection. Melatonin administered in the afternoon (15:00 h) was even more effective in stimulating the lowered TIDA neuronal activity and inhibiting the increased PRL level than that given in the morning (10:00 h). S-20098, a melatonin agonist was also effective in stimulating the TIDA neurons. In contrast, S-20928, a putative melatonin antagonist, while it had no effect by itself, blocked the effect of S-20098. Although S-20928 failed to prevent melatonin's effect on ME DOPAC levels, six interspaced injections of S-20928, from 18:00 to 01:30 h, significantly blocked the increase of ME DOPAC levels at 03:00 h, indicating that the endogenous melatonin may play a role. We further used rats that received daily injection of melatonin (1 mg/kg, ip) at 18:00 h for 10 days and found that the injection augmented basal TIDA neuronal activity at 11:00 h and blunted the afternoon PRL surge. In all, melatonin can have an inhibitory effect on PRL secretion by stimulating the TIDA neurons, and it may help to entrain the circadian rhythms of both TIDA neuronal activity and PRL secretion. PMID- 10831158 TI - Differential growth inhibitory effect of melatonin on two endometrial cancer cell lines. AB - The effect of melatonin on endometrial cancer cell growth was investigated using two cell lines, SNG-II and Ishikawa, which are different in their estrogen receptor status. A physiological concentration of melatonin (10(-9) M) showed no growth inhibitory effect on SNG-II cells, which are estrogen receptor-negative at all cell densities and incubation times. In contrast, melatonin significantly inhibited Ishikawa cells, which are estrogen receptor-positive at all cell densities tested after 96 hr incubation. The greatest inhibition of Ishikawa cell growth was observed at 10(-9) M melatonin, compared with other supra (10(-6), 10( 8) M) or subphysiological concentrations (10(-10), 10(-12) M). This growth inhibitory effect of melatonin on Ishikawa cells was completely blocked by 10( 10) to 10(-8) M concentrations of 17-beta estradiol administration. Pretreatment with luzindole, which is a selective melatonin receptor antagonist, prior to the addition of melatonin also blocked the inhibitory effect of melatonin on Ishikawa cells. This is the first study to demonstrate an anti-proliferative effect of physiological melatonin on endometrial cancer cells in vitro. The present study revealed that melatonin also inhibits the growth of estrogen receptor positive endometrial cancer cells and that this effect of the pineal indole may be mediated by both steroid and melatonin receptors. PMID- 10831159 TI - Different effects of melatonin pretreatment on cAMP and LH responses of the neonatal rat pituitary cells. AB - The rhythm of the pineal hormone melatonin transduces the effect of photoperiod on seasonal functions. Duration of the melatonin pulse provides information about season and the long melatonin pulse induces reproductive involution in the long day breeders such as photoperiodic rodents. The length of melatonin pulse thus carries photoperiodic information, which regulates the function of target cells. Therefore, we have studied the effects of melatonin pretreatment of various lengths on responsiveness of the neonatal rat pituitary cells cultured in vitro to GnRH or forskolin. In these cells, melatonin treatment inhibits the GnRH induced LH release as well as the forskolin-induced cAMP accumulation. However, long preincubation with melatonin has a paradoxical stimulatory effect on the cellular responsiveness. When the cells are pretreated with melatonin for 16 hr or more, then rinsed thoroughly and treated with forskolin for 30 min, the increase of cAMP is potentiated. Moreover, in the melatonin-pretreated cells. the subsequent melatonin treatment inhibits the forskolin-induced cAMP accumulation relatively more than in the non-pretreated cells. Although melatonin pretreatment does not potentiate the GnRH-induced LH release, it protects the gonadotrophs against the GnRH-induced desensitization: pretreatment with GnRH for 12 hr or more renders the cells insensitive to subsequent GnRH stimulation, while after pretreatment with GnRH and melatonin, the subsequent GnRH treatment induces significant increase of LH release. These observations indicate that long pretreatment with melatonin improves responsiveness of the pituitary cells to the subsequent stimulation, but its effects on cAMP accumulation and LH release are different. PMID- 10831160 TI - Melatonin-induced increased activity of the respiratory chain complexes I and IV can prevent mitochondrial damage induced by ruthenium red in vivo. AB - Melatonin displays antioxidant and free radical scavenger properties. Due to its ability with which it enters cells, these protective effects are manifested in all subcellular compartments. Recent studies suggest a role for melatonin in mitochondrial metabolism. To study the effects of melatonin on this organelle we used ruthenium red to induce mitochondrial damage and oxidative stress. The results show that melatonin (10 mg/kg i.p.) can increase the activity of the mitochondrial respiratory complexes I and IV after its administration in vivo in a time-dependent manner; these changes correlate well with the half-life of the indole in plasma. Melatonin administration also prevented the decrease in the activity of complexes I and IV due to ruthenium red (60 microg/kg i.p.) administration. At this dose, ruthenium red did not induce lipid peroxidation but it significantly reduced the activity of the antioxidative enzyme glutathione peroxidase, an effect also counteracted by melatonin. These results suggest that melatonin modulates mitochondrial respiratory activity, an effect that may account for some of the protective properties of the indoleamine. The mitochondria-modulating role of melatonin may be of physiological significance since it seems that the indoleamine is concentrated into normal mitochondria. The data also support a pharmacological use of melatonin in drug-induced mitochondrial damage in vivo. PMID- 10831161 TI - Differential effects of smoking on myocardial infarction risk according to the Gln/Arg 192 variants of the human paraoxonase gene. AB - Paraoxonase (PON1) seems to exert a major antioxidant effect by removing lipid peroxidation products. A common polymorphism of the PON1 gene modulates paraoxonase activity and has been related in some studies to coronary heart disease. PON1 genetic polymorphism includes PON1 Q, an isoform with a low activity toward paraoxon hydrolysis that has a glutamine at position 192, and PON1 R, the high-activity isoform with an arginine at position 192. In the present study, we investigated whether smoking, which is related to increased susceptibility to lipoprotein oxidation, has a differential effect by PON1-192 genotype on the risk of myocardial infarction (MI). One hundred fifty-six consecutive MI patients and 310 control subjects were studied. PON1 genotypes in the controls were distributed as follows: 154 (49.7%) QQ, 123 (39.7%) QR, and 33 (10.6%) RR. This distribution did not significantly differ from that of the MI patients: 84 (53.8%) QQ, 60 (38.5%) QR, and 12 (7.7%) RR. Subjects were classified into two groups, those who never smoked (n = 209) and those who were current smokers (n = 135) or ex-smokers (n = 122). In the latter, the variable "cigarette packs smoked per year" was defined as the number of packs smoked daily multiplied by the number of smoking years. As expected, smoking was significantly associated with an increased MI risk in the overall group. Subjects were then stratified by PON1 genotype. The packs smoked per year were significantly associated with an increased MI risk only in QQ homozygotes. This risk was higher among those in the higher tertile for cigarette packs smoked per year (odds ratio [OR] = 5.24, 95% confidence interval = 1.67 to 16.44, Pfor trend <.001). In contrast, the packs smoked per year were not significantly associated with MI risk in R-carrier subjects. We conclude that the risk of MI associated with smoking appears to be increased in subjects who are homozygous for the low activity PON1 QQ genotype compared with R carriers, and this risk seems to be time- and dose-dependent. PMID- 10831162 TI - Suppression by eicosapentaenoic acid of oxidized low-density lipoprotein and lysophosphatidylcholine-induced migration in cultured rat vascular smooth muscle cells. AB - The migration of medial smooth muscle cells into the intima is proposed to be an initial process of intimal thickening in atherosclerotic lesions. The present study was designed to determine whether pretreatment with the antiatherogenic agent eicosapentaenoic acid (EPA) inhibits the migration induced by oxidized low density lipoprotein (LDL) and its major phospholipid component, lysophosphatidylcholine (lyso-PC), in cultured rat vascular smooth muscle cells (VSMCs) using Boyden's chamber method. The effects of EPA pretreatment on angiotensin II (Ang II)- and platelet-derived growth factor BB (PDGF BB)-induced migration were also examined in these cells. Oxidized LDL and lyso-PC induced migration in a concentration-dependent manner. EPA pretreatment clearly suppressed oxidized LDL (200 microg/mL)- and lyso-PC (10(-5) mol/L)-induced migration between 40 and 160 micromol/L. EPA pretreatment also suppressed Ang 11 (10(-7) mol/L)- and PDGF BB (5 ng/mL)-induced migration at a concentration of 80 and 160 micromol/L. However, in a trypan blue exclusion test, dead cells stained with trypan blue were not found 24 hours after treatment with EPA. These results suggest that EPA suppresses VSMC migration induced by oxidized LDL and lyso-PC, as well as Ang II and PDGF BB. These preliminary data concerning the effects of EPA may partly explain the antiatherosclerotic effects of this agent. PMID- 10831163 TI - Effects of growth hormone treatment on very-low density lipoprotein apolipoprotein B100 turnover in adult hypopituitarism. AB - Adult hypopituitarism is associated with hyperlipidemia, mainly due to an increase of very-low-density lipoprotein (VLDL) and low-density lipoprotein (LDL) levels. Recent studies have shown that such patients exhibit increased hepatic secretion of VLDL apolipoprotein B100 (VLDL apo B100). To examine the effects of growth hormone (GH) replacement on VLDL apo B100 turnover, 13 GH-deficient hypopituitary patients (8 women and 5 men; aged 47 +/- 3 years, mean +/- SEM; body mass index [BMI], 30 +/- 2 kg/m2) entered a double-blind placebo-controlled study for 6 months (GH 0.125 IU/kg/wk for 4 weeks, and then 0.25 IU/kg/wk). GH was subsequently used in all patients for a further 6 months. A 6-hour [1-13C] leucine infusion was administered at baseline and at 6 months. The secretion rate of VLDL apo B100 was derived by kinetic analysis following quantitation of isotopic enrichment by gas chromatography/mass spectrometry. The GH-treated group (6 patients) demonstrated a similar fractional secretion rate (FSR) for VLDL apo B100 at 0 and 6 months. The pool size and absolute secretion rate (ASR) also were unaffected significantly by GH therapy. No significant changes were observed in the placebo group (7 patients). Treatment with GH for 6 months caused an increase in the high-density lipoprotein (HDL) cholesterol concentration (13 patients, 1.27 +/- 0.13 v 1.16 +/- 0.10 mmol/L, respectively, P = .05), whereas total cholesterol and triglyceride concentrations did not change. Nonesterified fatty acids (NEFAs) increased during GH therapy (471 +/- 43 micromol/L at 6 months v 349 +/- 49 micromol/L at baseline, P < .0005). The data suggest that GH does not affect VLDL apo B100 turnover in a significant way. PMID- 10831164 TI - Increased levels of advanced glycosylation end products in the kidney and liver from spontaneously diabetic Chinese hamsters determined by immunochemical assay. AB - Increased levels of advanced glycosylation end products (AGEs) have been reported in tissues in association with diabetes mellitus. Thus, we measured tissue AGE levels and detected an accumulation of AGEs in the kidney and liver from spontaneously diabetic Chinese hamsters (CHAD) to determine the relationship between AGEs and diabetes mellitus. Diabetic CHAD aged 12 to 13 months were studied together with age-matched nondiabetic CHAD. We used an AGE-specific noncompetitive enzyme-linked immunosorbent assay (ELISA) with polyclonal anti-AGE bovine serum albumin (BSA) antibody to measure tissue AGE levels. The samples extracted from the kidney and liver obtained from diabetic and nondiabetic CHAD reacted with anti-AGE-BSA antibody. When the absorbance of standard AGE-BSA (0.1 microg/mL) was expressed as 1 U, AGE levels in the kidney and liver from diabetic CHAD were significantly increased as compared with nondiabetic CHAD (kidney, 0.26 +/- 0.05 v 0.10 +/- 0.03 U/microg protein, P< .01; liver, 0.20 +/- 0.03 v 0.09 +/ 0.02 U/microg protein, P< .01). Positive AGE staining was observed in the renal cortex, especially in the tubules of diabetic CHAD, but little AGE staining was observed in the glomerulus by the immunohistochemical study. AGE staining was diffuse in the hepatocytes. These AGE levels were significantly correlated with fasting plasma glucose and glycated hemoglobin (P < .01, respectively). In conclusion, we have confirmed that AGE structures were expressed in the kidney and liver from CHAD, and these AGE levels were increased in diabetic CHAD. AGE staining was observed in the renal tubules and hepatocytes. Tissue AGE levels were positively correlated with glycemic control in CHAD. PMID- 10831165 TI - JTT-501, a new oral hypoglycemic agent, reverses hypertriglyceridemia in Zucker fatty and ventromedial hypothalamus-lesioned obese rats. AB - JTT-501 is a new oral hypoglycemic agent that is reported to be effective in insulin-resistant diabetic animal models by improving insulin resistance. It also improves hypertriglyceridemia. We investigated the mechanism of the reversal of hypertriglyceridemia in two types of obese animals using JTT-501. In Zucker fatty obese rats, an animal model of genetic obesity, fasting plasma triglyceride and glucose significantly decreased after a single daily oral dose of JTT-501 (100 mg/kg) for 7 days. In ventromedial hypothalamus (VMH)-lesioned obese rats, an animal model of nongenetic obesity, fasting plasma triglycerides significantly decreased but fasting plasma glucose levels remained unchanged after treatment with this agent. In Sprague-Dawley (SD) rats, fasting plasma triglyceride and glucose levels remained unchanged. The JTT-501-treated Zucker fatty and VMH lesioned obese rats showed a decrease in insulin, but it was not significant, while the treated SD rats showed a significant decrease in insulin. Postheparin plasma lipoprotein lipase (LPL) increased significantly in treated Zucker fatty obese and SD rats, but did not change in VMH-lesioned obese rats. The hepatic triglyceride secretion rate (TGSR) did not change in any species treated with JTT 501. There was a negative correlation between postheparin plasma LPL and plasma triglyceride levels in Zucker fatty obese rats, while no such correlation was observed in VMH-lesioned obese or SD rats. The fractional catabolic rate (FCR) for plasma triglyceride was increased significantly by JTT-501 in both Zucker fatty and VMH-lesioned obese rats. These results suggest that JTT-501 decreases plasma triglycerides mainly by increasing postheparin plasma LPL in Zucker fatty obese rats, while it ameliorates an impairment in the ability of adipose tissue to remove triglyceride from the circulation in VMH-lesioned obese rats. PMID- 10831166 TI - No difference in the lipolytic response to beta-adrenoceptor stimulation in situ but a delayed increase in adipose tissue blood flow in moderately obese compared with lean men in the postexercise period. AB - This study was undertaken to determine the effect of previous exercise on adipose tissue responsiveness to beta-adrenoceptor stimulation and on adipose tissue blood flow (ATBF). Eight lean and 8 obese men (body mass index [BMI], 23.6 +/- 2.1 [SD] v 29.0 +/- 1.9 kg x m(-2)) were investigated with abdominal subcutaneous microdialysis and 133Xe clearance. A stepwise isoprenaline infusion (10(-8), 10( 7), and 10(-6) mol x L(-1)) was administered in situ in the microdialysis catheter before and 2 hours after a submaximal exercise bout (90 minutes of cycling at 55% of maximal O2 uptake). No differences in the response (increase in interstitial glycerol v preinfusion level) to isoprenaline infusions were found between the 2 groups. In both groups, there was no difference in the response to postexercise versus preexercise infusion. When the vasodilating agent hydralazine (0.125 g x L(-1)) was infused into the microdialysis catheter to control for the vascular effects of isoprenaline, an interaction effect between exercise and isoprenaline dose was found. Analyses showed an attenuated response to the high isoprenaline dose after exercise (lean, 251 +/- 42 [SE] micromol x L(-1); obese, 288 +/- 77 micromol x L(-1)) versus before exercise (lean, 352 +/- 62 micromol x L(-1), P = .045 v after; obese, 380 +/- 94 micromol x L(-1), P = .021 v after), indicating a desensitization of lipolysis to beta-adrenoceptor stimulation. ATBF and arterial plasma glycerol increased after exercise in both groups, but the increase was delayed in obese subjects. Arterial plasma insulin was higher in the obese versus lean subjects at all times, and decreased during recovery in both groups. In conclusion, abdominal subcutaneous adipose tissue responsiveness to beta-stimulation is not enhanced postexercise in lean and obese men, whereas previous exercise increases ATBF. Furthermore, the data suggest slower lipid mobilization postexercise and resistance to the antilipolytic effect of insulin in the obese. PMID- 10831167 TI - Effect of the lipoprotein lipase activator NO-1886 on adriamycin-induced nephrotic syndrome in rats. AB - Hyperlipidemia associated with nephrotic syndrome may play a role in the deterioration of renal function. Tsutsumi et al have previously reported that the novel compound NO-1886 increases lipoprotein lipase (LPL) activity, resulting in a reduction of plasma triglycerides and an elevation of high-density lipoprotein (HDL) cholesterol in normal rats. The aim of this study was to ascertain whether NO-1886 suppresses the renal injury by treatment of the hyperlipidemia in an Adriamycin (Kyowa Hakko Kogyo, Tokyo, Japan) induced nephrosis rat model fed a high-protein diet that induced renal dysfunction and tubulointerstitial injury. Administration of Adriamycin caused hyperlipidemia, proteinuria, and edema with ascites in rats in 4 weeks. Furthermore, a combination of Adriamycin and a high protein diet increased plasma creatinine and blood urea nitrogen (BUN) and decreased plasma albumin. Histologically, in Adriamycin-treated rats, marked interstitial cellular infiltration, tubular lumen dilation, and tubular cast formation in the kidney were observed. NO-1886 decreased plasma triglyceride and increased HDL cholesterol in Adriamycin-induced nephrotic rats. NO-1886 treatment reduced plasma creatinine and BUN levels and increased plasma albumin in Adriamycin-treated rats; it also ameliorated the ascites and proteinuria. Histologically, NO-1886-treated rats showed a quantitatively significant preservation of tubulointerstitial lesions. These data suggest that NO-1886 may have a protective effect against Adriamycin-induced nephrosis with tubulointerstitial nephritis in rats by a modification of the plasma lipid disorder. PMID- 10831168 TI - Comparative study of the lipogenic potential of human and rat adipose tissue. AB - The reported low activity of lipogenic enzymes (especially adenosine triphosphate [ATP]-citrate lyase) in human adipose tissue led to the general conclusion that in humans lipogenesis occurs primarily in the liver. However, recent studies indicate that the liver plays a minor role in de novo lipogenesis and suggest that adipose tissue may be the principal lipogenic human tissue. In an attempt to resolve these contradictions we reinvestigated the lipogenic potential of human adipose tissue and compared with adipose tissue of rats fed a high-fat diet for 2 weeks and fasted overnight before death. These conditions mimic the nutritional state of patients at the moment of tissue sampling. We found that overnight fasting of the rats maintained previously for 12 days on a high-fat diet caused a decrease of ATP-citrate lyase of about 7-fold. Thus, in human adipose tissue, the mean activity of ATP-citrate lyase was approximately 8 times lower than in rats fed a high-fat diet and fasted overnight, and about 50 times lower than in rats maintained on normal laboratory diet. Unlike ATP-citrate lyase, fatty acid synthase (FAS) activity was only slightly lower in human adipose tissue than in rats maintained on a normal laboratory diet. Comparable FAS activity was found when rats were fed a high-fat diet and fasted overnight. The average activities of human adipose tissue acetyl-coenzyme A carboxylase, malic enzyme, and glucose 6-phosphate dehydrogenase were approximately 3-, 4-, and 6-fold lower than in adipose tissue from rats fed a high-fat diet and fasted overnight before tissue sampling, while the activity of 6-phosphogluconate dehydrogenase in humans was higher than in rat adipose tissue. No significant differences in lipogenic enzyme activities were found between male and female and between lean and obese patients. The rate of fatty acid synthesis in intact pieces of human adipose tissue was approximately 5 times lower than in adipose tissue pieces of rats fed a high-fat diet and fasted overnight before tissue samples were taken. The comparison of the lipogenic potential of humans and rats (maintained on the diet to mimic the nutritional state of patients at the time of tissue sampling) suggests that human adipose tissue is an important site of fatty acid synthesis. PMID- 10831169 TI - Impairment of skin vasoconstrictive response to sympathetic activation in obese patients: influence of rheological disorders. AB - Alterations of cardiac vagosympathetic activity have been suggested in obesity. We have previously shown that the skin vasoconstrictive response to sympathetic activation is reduced in non-insulin-dependent diabetic patients. The present study investigates the skin vasoconstrictive response to sympathetic activation in nondiabetic obese patients and the influence of clinical and rheological factors. Fifty-seven obese and 18 healthy women were investigated. The resting cutaneous blood flow (CBF) and CBF response to three tests that activate the sympathetic nervous system (deep breathing, Valsalva maneuver, and sitting to standing) were measured by a laser Doppler device. The red blood cell (RBC) filtration index (FI) and RBC aggregation were measured using a Hanss hemorrheometer and a Myrenne aggregometer (Myrenne, Roetgen, Germany), respectively. Resting CBF was not significantly different in obese and control subjects. The vasoconstrictive response to the deep-breathing and sitting-to standing tests expressed as the decrease in CBF was significantly lower in obese patients versus controls (43.9% +/- 3.1% v 73.7% +/- 17.9%, P = .01, and 67.1% +/ 3.8% v 89.8% +/- 12.0%, P = .02, respectively). The spontaneous basal CBF variations and the downward slope of the CBF reduction during the Valsalva and sitting-to-standing tests correlated negatively with age in obese patients (P = .042, .022, and .008, respectively). During the sitting-to-standing test, the percent change in CBF correlated positively with RBC aggregation at a shear rate of 0 and 3 s(-1) (P = .011 and .017, respectively). In conclusion, (1) CBF assessment by laser Doppler flowmetry is an effective noninvasive method to investigate sympathetic nervous function in obese patients; (2) obesity is associated with a significant reduction in the vasoconstrictive response to two tests for sympathetic activation, the deep-breathing and sitting-to-standing tests; (3) the severity of this reduction increases with age; and (4) RBC aggregation may contribute to the increase in the vasoconstrictive response and may thus increase the risk of widespread cardiovascular disease. PMID- 10831170 TI - Prevalence of paradoxically normal serum cholestrol in morbidly obese women. AB - The paradox that cholesterol may be lower in extremely obese subjects versus those who are less overweight, although originally observed more than 40 years ago, has never been documented in a systematic fashion. We have therefore prospectively determined the body mass index (BMI) and serum cholesterol concentration in 3,312 women. The percentage of women with serum cholesterol in the normal range (<200 mg/dL) decreased with an increasing BMI, from 55% in women with a BMI less than 20 kg/m2 to 28% in those with a BMI of 30 to 35 kg/m2. Serum cholesterol greater than 300 mg/dL was found in only 2% of individuals with a BMI less than 20 kg/m2 but in 6% of the group with a BMI between 30 and 35 kg/m2. However, among morbidly obese women (BMI >40 kg/m2, n = 46), 39% presented with serum cholesterol less than 200 mg/dL and only one woman had serum cholesterol more than 300 mg/dL. With the BMI, the fitted regression model shows an increase in cholesterol for low BMIs, while cholesterol appears to decrease with larger values for the BMI. The age-dependent increase in cholesterol is more evident in younger women versus older women, where it tends to disappear. It is concluded that among morbidly obese women (BMI >40 kg/m2), there is a substantial subgroup with normal serum cholesterol. PMID- 10831171 TI - Metabolic effects of insulin and insulin-like growth factor-I in endotoxemic rats during total parenteral nutrition feeding. AB - The effects of insulin and insulin-like growth factor-I (IGF-I) on protein, energy, and glucose metabolism were examined in endotoxemic rats receiving total parenteral nutrition (TPN) for 3 days. The endotoxemic model was induced by constant infusion of lipopolysaccharide (1 mg/kg x d) for 3 days. The TPN regimen provided 200 kcal/kg x d and 1.5 g protein/kg x d. The dosage of insulin (5 mU/kg x h) and IGF-I (20 microg/kg x h), either alone or in combination, was chosen to maintain normal levels of leucine and glucose in plasma during feeding. One normal control and 4 endotoxemic groups with different treatments (saline, IGF-I, insulin, or IGF-I and insulin) were included. The effects of endotoxin were compared between the group receiving endotoxin alone and normal controls, and the effects of insulin and IGF-I were compared within the endotoxemic groups. The results show that endotoxin significantly increased the mortality and induced a hypermetabolic state, and nutrition alone could not overcome the catabolism induced by endotoxin. However, administration of insulin and IGF-I enhanced protein preservation in muscle tissue in endotoxemic rats during TPN. This effect was greater for insulin either alone or in combination with IGF-I. Insulin also significantly reduced the mortality. There were no additive effects of these two anabolic hormones on any measured parameter in these experimental conditions. PMID- 10831172 TI - Relation of rat fetal body weight to the relative expression of insulin genes I and II. AB - This study investigates the relation between the ratio of the abundance of mRNAs encoded by insulin genes I and II (insulin I/II mRNA ratio) and the weight of rat fetuses on gestational day 20. Total RNA was extracted from the pancreas of the fetuses with the maximum and minimum body weight in each litter on gestational day 20. The amount of insulin mRNAs I and II in each RNA preparation was determined by reverse transcription-polymerase chain reaction (RT-PCR) analysis and restriction enzyme digestion. The maximum and minimum weight of the fetuses were 4.07 +/- 0.23 and 3.23 +/- 0.34 g, respectively (N = 18, P < .01) and the corresponding insulin I/II mRNA ratios were 3.65 +/- 0.55 and 1.42 +/- 0.21 (N = 18, P < .01). Furthermore, the insulin I/II mRNA ratio correlated significantly with fetal weight (r = .526, P < .05). These results suggest that the relative expression of insulin genes I and II may affect the extent of fetal growth. PMID- 10831173 TI - Effect of lysine infusion on urea cycle in lysinuric protein intolerance. AB - Poor intestinal absorption and excessive renal loss of dibasic amino acids result in low plasma concentrations in patients with lysinuric protein intolerance (LPI). Arginine and ornithine deficiency impair the function of the urea cycle and cause hyperammonemia after protein intake, while chronic lysine deficiency may cause growth failure and lead to reduced bone density in such patients. Since high lysine concentrations inhibit several enzymes of the urea cycle in the liver, lysine supplementation may induce hyperammonemia in LPI. We thus studied how LPI patients tolerate high plasma lysine by intravenous (IV) infusion of 3.3 mmol/kg lysine hydrochloride over 90 minutes in 6 adult patients and 4 healthy controls. The plasma lysine concentration (mean +/- SD, range) peaked in the patients (9,114 +/- 1,864, 7,156 to 12,044 micromol/L) and controls (10,185 +/- 2,253, 7,714to 13,122 micromol/L) at 90 minutes. Urinary lysine excretion peaked in the second 2-hour urine collection in the patients (4,582 +/- 1,276, 3,018 to 6,315 micromol/m2 body surface area per hour) and in the first 2-hour collection in the controls (5,373 +/- 1,766, 3,551 to 7,286 micromol/m2/h). Two patients had mild nausea but no hyperammonemia and one patient had moderate hyperammonemia (peak, 112 micromol/L) at the end of the infusion. Orotic acid excretion increased in 2 subjects with a peak excretion rate of 33 and 251 micromol/m2/h in the third 2-hour collection after starting the load. All other subjects remained asymptomatic and showed no change in plasma ammonia or urinary orotic acid excretion. We thus conclude that an acute increase in plasma lysine caused minimal clinical or biochemical untoward effects in patients with LPI. Moderate increases in plasma lysine after low-dose oral supplementation with lysine or well-absorbed lysine derivatives are probably well tolerated in LPI. PMID- 10831174 TI - Insulin-like growth factor binding proteins (IGFBPs) in serum and urine and IGFBP 2 protease activity in patients with insulin-dependent diabetes mellitus. AB - Diabetes mellitus and glucose dysregulation have significant effects on the circulating level of insulin-like growth factor-I (IGF-I) and IGF binding proteins (IGFBPs). In the present study, serum and urine IGFBP (IGFBP-1, -2, and 3) and serum IGF-I and -II levels were measured by radioimmunoassay (RIA) in 27 patients with type 1 diabetes aged 9 to 48 years compared with 9 healthy subjects aged 10 to 28 years. The patients were divided into 3 groups according to the amount of albumin excreted in 24 hours. The macroalbuminuria group (>500 mg/24 h) had elevated serum IGFBP-1 and -2 and decreased IGF-I levels (P < .01 v normal controls). Serum IGFBP-3 and IGF-II were not different among the patient groups and controls (P > .05). The mean urinary IGFBP-1 was decreased in all 3 patient groups compared with the controls (P < .05). Urinary IGFBP-2 and IGFBP-3 were increased in patients with macroalbuminuria. Immunoblot analysis showed increased low-molecular-weight fragments of urinary IGFBP-2 in the poorly controlled diabetics, and direct evidence for increased urinary IGFBP-2 proteolytic activity could be demonstrated in both the microalbuminuric and macroalbuminuric groups. Low-molecular-weight fragments of urinary IGFBP-3 were also increased in both the microalbuminuric and macroalbuminuric groups. In conclusion, alterations of IGFBPs in urine and serum are related to metabolic control in diabetic patients, and there is an increase of urinary IGFBP-2 protease activity in poorly controlled diabetics. The changes in serum IGFBP concentrations (eg, increases in IGFBP-1 and IGFBP-2) may lead to alterations in the availability of IGF-I to peripheral tissues. PMID- 10831175 TI - Exercise-induced microalbuminuria in patients with active acromegaly: acute effects of slow-release lanreotide, a long-acting somatostatin analog. AB - Recent clinical studies have demonstrated an increase of urinary albumin excretion (UAE) at rest in acromegalic patients and, on the other hand, a reduced UAE in patients with growth hormone (GH) deficiency. Physical exercise is known to induce abnormal UAE in patients with diabetes, probably unmasking early glomerular alterations. The effect of exercise on UAE in acromegaly is not known. Moreover, the effect of acute but sustained GH inhibition in acromegaly on UAE at rest and after exercise has never been studied. The aim of our study was to evaluate the acute short-term effects of slow-release lanreotide (SR-L), a long acting somatostatin analog, on UAE and alpha1-microglobulinuria (A-1-M), a marker of renal tubular damage, at rest and after exercise in 7 normotensive patients with active acromegaly and normal renal function (4 males and 3 females; mean age, 53 +/- 3.1 years; body mass index [BMI], 27.3 +/- 1.1 kg/m2) at baseline and 7 and 14 days after SR-L injection (30 mg). Two of the acromegalic patients were microalbuminuric at rest, and in other 3 cases, UAE was in the borderline range (10 to 20 microg/min). At baseline in the acromegalic subjects, we found a significant increase in UAE at rest with respect to 7 normal subjects considered as a control group. GH and insulin-like growth factor-1 (IGF-1) were also reduced compared with baseline 7 and 14 days after SR-L injection (GH, 13.4 +/- 7.3 and 13.61 +/- 7 v 18.5 +/- 9.3 microg/L, P < .05; IGF-1, 230 +/- 53 and 255 +/- 54 v 275 +/- 64 microg/L). Concomitantly, we observed a significant decrease of UAE at rest and after exercise and 7 and 14 days after SR-L injection as compared with baseline values (27.3 +/- 20.5 and 18.2 +/- 13.7 v 35.3 +/- 12.8 microg/min, P < .05; exercise, 48.5 +/- 24.1 and 18.6 +/- 6.8 v68.3 +/- 39.7 microg/min, P < .05). A-1-M always remained in the normal range (< 12 mg/L) both at rest and after exercise. We can thus conclude that in acromegaly, submaximal exercise induces abnormal increases in microalbuminuria. We hypothesize that this phenomenon may be due to the functional glomeruler involvement. SR-L can significantly reduce UAE at rest and after exercise in the short-term in acromegaly, probably via a decrease in circulating GH levels. PMID- 10831176 TI - Effect of physical exercise on lipoprotein(a) and low-density lipoprotein modifications in type 1 and type 2 diabetic patients. AB - To evaluate the effect of physical exercise on blood pressure, the lipid profile, lipoprotein(a) (Lp(a)), and low-density lipoprotein (LDL) modifications in untrained diabetics, 27 diabetic patients (14 type 1 and 13 type 2) under acceptable and stable glycemic control were studied before and after a supervised 3-month physical exercise program. Anthropometric parameters, insulin requirements, blood pressure, the lipid profile, Lp(a), LDL composition, size, and susceptibility to oxidation, and the proportion of electronegative LDL (LDL( )) were measured. After 3 months of physical exercise, physical fitness improved (maximal O2 consumption [VO2max], 29.6 +/- 6.8 v 33.0 +/- 8.4 mL/kg/min, P < .01). The body mass index (BMI) did not change, but the waist circumference (83.2 +/- 11.8 to 81.4 +/- 11.2 cm, P < .05) decreased significantly. An increase in the subscapular to triceps skinfold ratio (0.91 +/- 0.37 v 1.12 +/- 0.47 cm, P < .01) and midarm muscle circumference ([MMC], 23.1 +/- 3.4 v 24.4 +/- 3.7 cm, P < .001) were observed after exercise. Insulin requirements (0.40 +/- 0.18 v 0.31 +/ 0.19 U/kg/d, P < .05) and diastolic blood pressure (80.2 +/- 10 v 73.8 +/- 5 mm Hg, P < .01) decreased in type 2 diabetic patients. High-density lipoprotein cholesterol (HDL-C) increased in type 1 patients (1.48 +/- 0.45 v1.66 +/- 0.6 mmol/L, P < .05), while LDL cholesterol (LDL-C) decreased in type 2 patients (3.6 +/- 1.0 v3.4 +/- 0.9 mmol/L, P < .01). Although Lp(a) levels did not vary in the whole group, a significant decrease was noted in patients with baseline Lp(a) above 300 mg/L (mean decrease, -13%). A relationship between baseline Lp(a) and the change in Lp(a) (r = -.718, P < .0001) was also observed. After the exercise program, 3 of 4 patients with LDL phenotype B changed to LDL phenotype A, and the proportion of LDL(-) tended to decrease (16.5% +/- 7.4% v 14.0% +/- 5.1%, P = .06). No changes were observed for LDL composition or susceptibility to oxidation. In addition to its known beneficial effects on the classic cardiovascular risk factors, regular physical exercise may reduce the risk of cardiovascular disease in diabetic patients by reducing Lp(a) levels in those with elevated Lp(a) and producing favorable qualitative LDL modifications. PMID- 10831177 TI - Modulation of plasma endothelin levels by the menstrual cycle. AB - Blood pressure varies during the menstrual cycle, but the reason for this is unclear. Administration of (synthetic) sex hormones can influence the level of vasoactive substances such as endothelin (ET). However, it is not known whether short-term variations in sex hormone levels in physiological situations affect ET levels. We assessed the effects of the menstrual cycle on plasma ET-1 in 8 healthy premenopausal women not using oral contraceptives (OCs) and 8 premenopausal women using OCs. ET-1 levels were measured in all subjects on days 1 to 3 (menstrual phase), 9 to 12 (follicular phase), and 20 to 23 (luteal phase) of the menstrual cycle. ET-1 levels remained constant in OC users (2.4 +/- 0.4, 2.6 +/- 0.4, and 2.4 +/- 0.4 pg/mL on days 1 to 3, 9 to 12, and 20 to 23 of the pill cycle). In contrast, ET-1 levels in non-OC users decreased in all women during the follicular and luteal phase of the menstrual cycle compared with the menstrual (low-estrogenic) phase (3.6 +/- 0.5, 2.8 +/- 0.5, and 2.9 +/- 0.3 pg/mL for the menstrual, follicular, and luteal phase, respectively, P < .01 for menstrual vfollicular and P < .01 for menstrual v luteal). The differences between OC users and nonusers were significant in the menstrual phase of the cycle (P < .01). We conclude that ET levels fluctuate during the menstrual cycle. Previously reported effects of the menstrual cycle on blood pressure may be partly explained by the effects of sex hormones on the level of vasoactive mediators. This fluctuation is not present in OC users. Studies on hemodynamic parameters in premenopausal women should account for hormonal variations in the various phases of the menstrual cycle. PMID- 10831178 TI - Paternal-maternal effects on phenotypic characteristics in spontaneously diabetic Nagoya-Shibata-Yasuda mice. AB - The Nagoya-Shibata-Yasuda (NSY) mouse is an inbred strain with spontaneous development of type 2 (non-insulin-dependent) diabetes mellitus. The purpose of this study was to determine the mode of inheritance of various phenotypes related to diabetes in this strain. Two reciprocal outcrosses, female C3H/He x male NSY F1 (C3NF1) and female NSY x male C3H/He F1 (NC3F1) mice, were performed. The phenotypic characteristics in both F1 mice were investigated. The cumulative incidence of diabetes was 100% (25 of 25) in male C3NF1 mice and 97% (29 of 30) in male NC3F1 mice at 48 weeks of age, indicating that diabetes in NSY mice was transmitted to male F1 hybrids in an autosomal dominant manner. Fatty liver also showed an autosomal dominant mode of inheritance. In contrast, epididymal fat accumulation and impaired insulin secretion showed an autosomal recessive mode of inheritance. The body mass index (BMI) showed a codominant mode of inheritance. Paternal-maternal effects associated with the severity of diabetes were observed. Insulin resistance was much more severe in male F1 mice than in the parental NSY strain. These data indicate different modes of inheritance among phenotypes related to type 2 diabetes. The presence of more severe insulin resistance in F1 mice versus the parental strains suggests the interaction of both parental genomes in the development of insulin resistance. The F1 mouse is expected to be useful for studies of the pathogenesis and genetic synergism of the insulin resistance syndrome. PMID- 10831179 TI - Treatment with diazoxide causes prolonged improvement of beta-cell function in rat islets transplanted to a diabetic environment. AB - Prolonged hyperglycemia desensitizes beta cells. A role for hyperglycemia-induced excessive stimulation can be tested by diazoxide, which inhibits glucose-induced insulin secretion. Using diazoxide, we have investigated in a rat transplantation model whether excessive stimulation can induce lasting effects on beta cells. One batch with 150 islets and another with 20 islets isolated from Wistar-Furth rats were transplanted under the left-kidney capsule of syngeneic streptozotocin diabetic recipients. In a first series, recipients were treated for 8 weeks with or without 0.2% diazoxide in the food. Graft-bearing kidneys were then perfused and excised. Diazoxide treatment increased by 5.5-fold the insulin response to 10 mmol/L arginine, by 4.1-fold the graft insulin content, and by 2.3-fold the preproinsulin mRNA versus nontreated diabetic controls. The persistence of these effects was assessed in a second series in which 8 weeks of diazoxide treatment was followed by 1 week of no treatment. Again, perfusion experiments showed a higher insulin response to arginine in diazoxide-treated rats (136.0 +/- 25.7 v 62.3 +/- 11.8 fmol/min, P < .05). Also, the response to 27.8 mmol/L glucose was increased (54.0 +/- 17.1 v 13.6 +/- 7.8 fmol/min, P < .05). The insulin content was increased (2.2 +/- 0.6 v 1.0 +/- 0.4 pmol/islet, P < .05), as well as the preproinsulin mRNA (0.60 +/- 0.08 v0.22 +/- 0.02 pg/islet, P < .05). In a third series, we tested the impact of diazoxide treatment when given only during the first 2 weeks following transplantation. When tested 6 weeks later, insulin secretion was unaffected, whereas there was a strong tendency for a higher preproinsulin mRNA and insulin content in grafts of diazoxide-treated rats. In conclusion, this study demonstrates that beta-cell function in transplanted islets is improved by diazoxide long after the end of treatment, an effect that is likely due to removal of hyperglycemia-induced excessive stimulation. PMID- 10831180 TI - Improvement by the insulin-sensitizing agent, troglitazone, of abnormal fibrinolysis in type 2 diabetes mellitus. AB - This study evaluated abnormal fibrinolysis in diabetic patients in terms of the pathophysiological significance and reversibility by oral hypoglycemic agents. Forty-seven patients with type 2 diabetes mellitus were randomly treated for 4 weeks with glibenclamide (n = 23) or troglitazone (n = 24). Before and after treatment, glycemic control, steady-state plasma glucose and insulin (SSPG and SSPI, respectively), and markers of fibrinolysis (tissue plasminogen activator [tPA] and plasminogen activator inhibitor-1 [PAI-1]) were analyzed in each patient. Pretreatment plasma PAI-1 in diabetic patients, but not tPA, was well correlated with the severity of retinopathy assessed by the fluorescence technique. Four weeks of treatment with troglitazone significantly decreased hemoglobin A1c (HbA1c), SSPG, and PAI-1 without an alteration of tPA. The troglitazone-induced decrease in plasma PAI-1 (50.3 v28.8 micromol/L; P < .05) was correlated with HbA1c (8.80% v7.21%, r = .539, P < .01) and SSPG (16.2 v 8.97 mmol/L, r = .562, P < .01) but not with SSPI. In contrast, treatment with glibenclamide for 4 weeks also reduced the HbA1c titer to almost the same extent as troglitazone (1.38% v 1.59%), but did not change the plasma PAI-1 or SSPG titer. These results suggest that an abnormal fibrinolytic state, especially overproduction of PAI-1, may be a pathogenic factor in the development of diabetic complications such as retinopathy, which may be improved by correction of the insulin resistance with troglitazone. PMID- 10831181 TI - Effect of tumor necrosis factor alpha and transforming growth factor beta 1 on plasminogen activator inhibitor-1 secretion from subcutaneous and omental human fat cells in suspension culture. AB - Elevated levels of plasminogen activator inhibitor-1 (PAI-1) are characteristic of the obese state and may contribute to the association between obesity and cardiovascular disease. In this study, we measured the rate of secretion of PAI-1 antigen in isolated subcutaneous and omental abdominal adipocytes from severely obese and non-obese individuals and studied the effect of selected cytokines on PAI-1 release using a suspension culture technique. PAI-1 secretion was approximately 2-fold greater in isolated fat cells from the obese versus non obese subjects. In addition, PAI-1 mRNA levels were higher in adipose tissue samples from obese versus non-obese individuals (P < .05). PAI-1 release was also approximately 2-fold greater in omental versus subcutaneous adipocytes from both obese and non-obese subjects (each P < .05). A 24-hour exposure to 1 nmol/L tumor necrosis factor alpha (TNF-alpha) slightly increased PAI-1 release from both subcutaneous and omental adipocytes (30% +/- 21% and 17% +/- 18%, respectively, nonsignificant [NS]). Transforming growth factor beta 1 (TGF-beta1) induced a significant dose-dependent increase of PAI-1 release into the medium. Exposure to 400 pmol/L TGF-beta1 of subcutaneous and omental fat cells from both obese and non-obese individuals elevated PAI-1 secretion by 2-fold. These data provide evidence that human fat cells release a substantial amount of PAI-1 in a depot specific manner and that TGF-beta1 particularly contributes to the regulation of PAI-1 secretion. PMID- 10831182 TI - Infantile obesity: a situation of atherothrombotic risk? AB - Obesity is a major risk factor for cardiovascular disease frequently associated with hypertension, dyslipidemia, and diabetes. In recent years, alterations in the hemostatic system have been added to these dysfunctions. We analyzed some of these alterations in coagulation and fibrinolysis in obese children (6 to 9 years old) of both sexes. We studied 61 obese children (mean body mass index [BMI], 22.35 kg/m2; 95% confidence interval [CI], 21.82 to 22.87) and 70 non-obese children (mean BMI, 16.58 kg/m2; 95% CI, 16.24 to 16.93) as a control group. The obese subjects presented significantly elevated values for insulin (P < .001), tissue-plasminogen activator ([t-PA] P < .001), plasminogen activator inhibitor-1 ([PAI-1] P < .001), and fibrinogen (P < .001) with respect to the control group. We found no significant differences in the concentration of glucose and fragment 1 + 2 of prothrombin (F1 + 2). In the obese subjects, insulin, PAI-1, and F1 + 2 were positively correlated with the BMI. On the other hand, t-PA was correlated with insulin and PAI-1 but not with the BMI. Therefore, in the obese children, there was an increment of the risk factors for cardiovascular disease. PMID- 10831183 TI - Regulation of splanchnic and renal substrate supply by insulin in humans. AB - To determine the effects of peripheral insulin infusion on total, hepatic, and renal glucose production and on the percent contribution to glucose production of gluconeogenesis versus glycogenolysis, 10 healthy subjects had arterialized hand and hepatic vein catheterization after an overnight fast and the results were compared with data from 12 age- and weight-matched subjects with renal vein catheterization during a 180-minute infusion of either insulin (0.25 mU/kg x min) with dextrose, or saline. Endogenous, hepatic, and renal glucose production was measured with [6,6(-2)H2]glucose, regional lactate, alanine, and glycerol balance by arteriovenous difference; hepatic blood flow by indocyanine green clearance; and renal blood flow by p-aminohippurate clearance, before and every 30 minutes during each infusion period. Insulin increased from about 42 to 98 pmol/L and blood glucose remained constant in all studies (3.8 +/- 0.2 v4.4 +/- 0.1 micromol/ml, hepatic vrenal vein). In response to insulin infusion, endogenous, hepatic, and renal glucose production decreased immediately (30 minutes) and reached a lower plateau value (10.8 +/- 0.8 v6.4 +/- 0.7, 10.4 +/- 1.1 v7.8 +/- 1.0, and 2.8 +/- 0.6 v 1.5 +/- 0.6 micromol/kg x min, respectively) between 120 and 180 minutes (all P < .05). Net renal uptake of lactate (2.4 +/- 0.4 v0.9 +/- 0.6) decreased earlier (30 minutes) and returned to baseline between 120 and 180 minutes (2.4 +/- 0.5 micromol/kg x min), whereas net splanchnic uptake of lactate (5.7 +/- 0.7 v 0.7 +/- 0.6) and alanine (1.8 +/- 0.1 v 1.0 +/- 0.5 micromol/kg x min) decreased later (120 to 180 minutes). Net renal (0.3 +/- 0.1 v 0.1 +/- 0.1) and splanchnic (0.7 +/- 0.3 v 0.4 +/- 0.2 micromol/kg x min) glycerol uptake decreased 90 to 180 minutes after insulin and increased (P < .05) with saline infusion (0.4 +/- 0.1 v0.6 +/- 0.3 and 1.0 +/- 0.5 v1.8 +/- 0.4 micromol/kg x min, respectively). These data indicate that the rapid suppression of endogenous glucose production by insulin reflects primarily a decrease in hepatic glucose release, most likely due to inhibition of net glycogenolysis, combined with suppression of renal gluconeogenesis. Inhibition of hepatic gluconeogenesis presumably occurs later during hyperinsulinemia. We conclude that peripheral insulin, in addition to its inhibition of glycogen degradation, regulates endogenous glucose production, in part, by modifying the splanchnic and renal substrate supply. PMID- 10831184 TI - Profiling of Zucker diabetic fatty rats in their progression to the overt diabetic state. AB - Blood chemistry profiles (glucose, insulin, and triglycerides) and indirect calorimetry were performed on male Zucker diabetic fatty (ZDF) rats in a longitudinal fashion (starting at 7 weeks of age) to assess the nature and timing of specific events in the transition to overt diabetes. Peripheral (skeletal muscle) insulin resistance was clearly present at 7 weeks of age in ZDF rats, yet circulating glucose was only slightly above normal as a result of compensatory hyperinsulinemia. At a crucial stage from 7 to 8 weeks, a reduction in insulin levels instigated several deleterious changes resulting in reduced whole-body carbohydrate utilization and increased glycemia. In subsequent weeks, an inability to sustain peripheral glucose disposal as a consequence of a continuous decline in insulin levels further reduced carbohydrate utilization (increased lipid utilization) and enhanced the overt hyperglycemia. These observations document in a systematic fashion the alterations that define diabetic progression in ZDF rats. PMID- 10831185 TI - Primary hyperparathyroidism in women: a tale of two cities--New York and Beijing. AB - The Western world has been used to describing disease on its terms, as if it is a prototype for the same disease found anywhere else in the world. It is unusual that one can test the hypothesis that a common disease can present in markedly different ways, depending on the country in which it is studied. We have had the opportunity to compare and contrast primary hyperparathyroidism in the United States and in China as seen in New York City and in Beijing. The cohort of subjects in each case was well over 100, and the experience extends to well over a decade of observations. In the United States, primary hyperparathyroidism typically presents as asymptomatic hypercalcemia in women within 10 years of menopause. Most often, it is discovered accidentally in the course of a routine multichannel chemistry screening test. The serum calcium is 10.5 + 0.1 mg/dL, within 1 mg/dL above the upper limit of normal, 10.2; the serum parathyroid hormone level is 118 + 9 pg/mL (within 1.5-2-fold above the normal limit, 65). The average 25-hydroxyvitamin D level is 21 ng/mL, in the lower range of normal. The classical clinical manifestations of primary hyperparathyroidism, stone and bone disease, have become much less common than earlier descriptions of the disease in the United States through the 1950s. Overt radiological bone disease (osteitis fibrosa cystica) is almost never seen, whereas stone disease is reduced in incidence from a high of 60% in the 1940s to current estimates of 15-20% now. Most patients are asymptomatic; skeletal involvement is detected only by measuring skeletal calcium by bone densitometry. Primary hyperparathyroidism in China presents much differently. Patients are younger, with an average age of 37. The serum calcium level is much higher, averaging about 12 mg/dL. PTH is over 20 times the upper limits of normal. The average 25-hydroxyvitamin D concentration is much lower than in the United States population, 8.8 ng/mL. Radiological evidence for osteitis fibrosa cystica is seen in 60% of patients; virtually all patients have osteoporosis. Thirty-five percent of patients suffer pathological fractures, most often of the femur or humerus. Forty-two percent demonstrate kidney stones, with half showing bilateral disease. Constitutional features of weakness and easy fatigability are always present. There are both facile and rather subtle explanations for this dramatically different presentation of the same disease in the United States (New York City) and China (Beijing). PMID- 10831186 TI - Cardiovascular issues with oral contraceptives: evidenced-based medicine. AB - Recent studies of current oral contraceptives indicate that the risk of cardiovascular sequelae is low in young (age 20-24 years) reproductive-aged women. Venous thromboembolism remains the one event that occurs in users independent of the presence of risk factors. However, the attributable risk is small, in the range of 7 to 18 events per 100,000 women annually. This risk is proportional to estrogen dose until the level of 30-35 microg is reached; type of progestin may also influence risk, though recent studies are controversial. Modifiable risk factors for venous thromboembolism include the presence of hemostatic disorders, especially factor V Leiden, and perhaps obesity. Stroke is even more uncommon, with an attributable risk of about 1.5 events per 100,000 women annually. Cigarette smoking and hypertension are modifiable risk factors for both ischemic and hemorrhagic stroke; use of preparations with 50 microg of estrogen or higher and migraine headaches are risk factors for ischemic stroke. Eliminating risk factors among users substantially reduces the risk of ischemic stroke and virtually eliminates the risk of hemorrhagic stroke. Myocardial infarction is rare among young women, occurring at a rate of about 0.2 event per 100,000 women annually. Oral contraceptive users who are non-smoking and normotensive do not have an increased risk of myocardial infarction. However, the presence of these risk factors along with age acts synergistically to increase the risk among oral contraceptive users. PMID- 10831187 TI - Skin breakthroughs in the year 2000. AB - Aging skin is a factor of many things-genetics, sun exposure, environmental insults, stress, and more. The signs of aging skin and cancer will be briefly reviewed followed by a discussion of current advances in dermatology, beginning with the new FDA regulation of sunscreens, the current state of tretinoin (Renova) and its future uses, the current state of antioxidants in the skin, and the latest therapies of microdermabrasion, neodymium:YAG lasers, and botulinum toxin. PMID- 10831188 TI - The menopause in Europe. AB - Most women in developed countries will live a third of their lives after the menopause. Vasomotor symptoms (hot flushes, night sweats, irritability, sleep disturbances, mood swings), and urogenital complications (atrophic vaginal irritation and dryness, dyspareunia) occur frequently during this period of life, but their severity and duration may vary widely between individuals. The menopause also induces accelerated bone loss and is the principal risk factor for osteoporosis. Hormone replacement therapy (HRT; estrogen or estrogen plus progestogen) alleviates these symptoms and can be administered orally, transdermally, topically, intranasally, or as subcutaneous implants. HRT is also effective for prevention and treatment of postmenosausal osteoporosis throughout the time that it is used. It is not surprising that HRT use has increased substantially during the past decade. Nevertheless, there are still considerable variations in use between different countries within the European community. This presentation will analyze: the frequency of menopausal symptoms among women in different European countries and the factors that influence them; the frequency of other postmenopausal women's health issues in Europe; the use of HRT in Europe as well as the type of HRT and its evolution during the last decade; and possible reasons explaining heterogeneity between countries. PMID- 10831189 TI - Expanding contraceptive choices: lunelle monthly contraceptive injection. AB - Contraceptive options for women have expanded with the FDA approval of a monthly injectable contraceptive. Containing a progestin and estrogen (25 mg medroxyprogesterone acetate and 5 mg estradiol cypionate; Lunelle/Lunella; Pharmacia and Upjohn), this novel once-a-month injection is characterized by the convenience and contraceptive efficacy of long-term reversible methods and the cycle control, return to fertility and side-effect profiles common to low-dose oral contraceptives. The association of these desirable attributes may increase the likelihood that a woman's chosen method will be used consistently and correctly to reduce the frequency of unintended pregnancy. PMID- 10831190 TI - Antiretroviral combination therapy in HIV-1 infected women and men: are their responses different? AB - Women are the fastest growing segment of the AIDS cases in the United States. They constitute nearly half of all the AIDS cases worldwide. Recent advances in Highly Active Antiretroviral Therapies (HAART) have reduced AIDS mortality remarkably. But as longer use of these combination regimens makes evident, unexpected side effects are now reported that might reflect gender-based differences in occurrence. Controversy still exists in relation to the level of HIV-1 quantification in men and women and its association with disease progression. Women have been reported to have lower viral loads with equal progression or higher progression with equal viral loads. This finding has not been consistent in all studies. Psychosocial variables, such as poverty, lack of care and young age, adversely affect more women than men. If the viral dynamics are thought to be different, then the response to treatment might be as well. So far, the effectiveness of HAART has been seen equally among men and women. Barriers to adherence, such as caregiving burdens, multiplicity of roles and fear of disclosure, might disproportionately affect women. By far the best news is that the survival of both men and women has improved with the newer therapeutic advances. PMID- 10831191 TI - Arrhythmias during spinal anesthesia. PMID- 10831192 TI - Surgery as team endeavour. PMID- 10831193 TI - Arrhythmias during spinal anesthesia for Cesarean section. AB - PURPOSE: Spinal block has long been considered a safe anesthesia technique for surgery. However, severe bradycardia, cardiac arrest, and other arrhythmias during spinal anesthesia have been reported and the incidence of intraoperative arrhythmias is not well established. In this study the incidence of arrhythmias during spinal anesthesia was determined. METHODS: We studied 254 healthy women undergoing Cesarean section under spinal anesthesia prospectively. Spinal anesthesia with 10 mg bupivacaine mixed with 0.2 mg morphine was performed at the L3-4 interspace. Intraoperative arrhythmias were recorded and verified later by a cardiologist. RESULTS: First degree atrioventricular block developed in nine patients (3.5%), second degree atrioventricular block in nine (3.5%), severe bradycardia (heart rate < 50 beats x min(-1)) in seventeen (6.7%), multiple VPC in three (1.2%). The height and weight of patients with severe bradycardia, multiple VPCs, or atrioventricular block were not different from those of the other patients. However, the age of patients in the potentially dangerous arrhythmias group was greater than that in the other group (P = 0.006). CONCLUSION: The incidence of arrhythmias as well as hypotension during spinal anesthesia for Cesarean section was higher than expected. Although most of these arrhythmias were transient and recovered spontaneously, they might unexpectedly occur and sometimes need immediate and prompt treatment. It is necessary to remain vigilant during spinal anesthesia for Cesarean section and careful monitoring of these patients is warranted, especially in older parturients. PMID- 10831194 TI - Clinimetric scale to measure surgeons' satisfaction with anesthesia services. AB - PURPOSE: Continuous quality improvement is of increasing interest to anesthesiologists. Since surgeons are coworkers and important clients of anesthesiologists, the level of satisfaction of surgeons with anesthesia services should be explored to optimize quality. The purpose of this study was, first, to introduce the concept of surgeons as clients of anesthesiologists and second, to develop and test an instrument to measure surgeons' satisfaction, the Surgeon Satisfaction with Anesthesia Services (SSAS) scale. METHODS: A conceptual model of surgeon satisfaction with anesthesia services was created before the development of the SSAS scale. The scale, composed of socio-demographic, Likert type and open-ended questions was sent to a sample of 250 surgeons selected randomly by the College des Medecins du Quebec. Exploratory factorial analysis were performed on the results. RESULTS: A Cronbach's alpha of 0.84 was obtained for internal consistency. Exploratory factorial analysis yielded two subscale factors: a) clinical expertise and b) attitudes and behaviour Global mean of surgeons'satisfaction was moderately high (3.11/4.0). Satisfaction was not related to sociodemographic variables. Very high scores were obtained for items related to clinical expertise. Items related to attitudes and behaviour obtained lower scores. A significant difference was obtained between both factors (t = 5.732, P = 0.0001). CONCLUSION: The SSAS scale is a new instrument to evaluate surgeon satisfaction. Overall, surgeons seem satisfied with anesthesia services, but many areas of dissatisfaction persist. Further discussions with surgeons should be encouraged, in view of improving the perceptions of the quality of anesthesia services and interprofessional relationships. PMID- 10831195 TI - Improved bowel function after gynecological surgery with epidural bupivacaine fentanyl than bupivacaine-morphine infusion. AB - PURPOSE: To compare postoperative gastrointestinal recovery between continuous epidural bupivacaine-fentanyl and bupivacaine-morphine. METHODS: In a blinded, randomized, prospective trial, 60 women undergoing surgery for gynecologic cancer were studied. Anesthesia was provided by a combined general/epidural (L2-3 catheter) technique without epidural opioids. Postoperative epidural analgesia was by continuous infusion of bupivacaine 0.1% with either morphine 0.05 mg x ml( 1) (BM) or fentanyl 5 microg x ml(-1) (BF). Visual Analogue Scale (VAS) scores for pain at rest and during movement, and the return of bowel function were collected for three days and the duration of hospitalization were noted. RESULTS: On POD-1, 18.5% of patients in the BM group had emesis compared with none in the BF group (P = 0.038) and fewer patients in the BM group tolerated clear oral fluids (11.1% BM vs 40.6% BF, P = 0.025). These differences became insignificant on POD-2 and 3. Median pain scores were comparable at rest and ranged from 10-20 in the BM group vs 0-20 in the BF group over the three days. Similarly, median pain scores with movement respectively ranged from 20-25 and 20-30 in the BF and BM groups. The mean duration of hospitalization was longer in the BM group (5.7 +/- 2.4) vs BF (4.5 +/- 1.2 days), P = 0.017. CONCLUSION: Epidural BM and BF provided equally effective postoperative analgesia at rest and during movement. Compared with BM, epidural BF is associated with less emesis and an increased ability to tolerate oral fluids on POD-1 and an overall shorter hospital stay. PMID- 10831196 TI - Lidocaine added to a tracheostomy tube cuff reduces tube discomfort. AB - PURPOSE: To examine whether lidocaine diffusion across an endotracheal tube cuff affects tracheostomy tube discomfort. METHODS: Two tracheostomy tube cuffs were inflated with 5 ml lidocaine 4% solution and air at 20 cmH2O, and then placed in 20 ml distilled water at 37 degrees C. After vigorous stirring, 100 microl of this water was then sampled immediately then 1, 2, 4, 8, 24 hr later to measure lidocaine concentration by high-performance liquid chromatography. Sixteen patients undergoing tracheostomy following oral cancer resection were randomly assigned to two groups: lidocaine (n=8) and placebo (n=8). A tracheostomy tube cuff was inflated with 5 ml lidocaine 4% or saline 0.9% and air to a cuff pressure of 20 cmH2O, in the lidocaine and placebo groups respectively. Tube discomfort was evaluated using a visual analogue scale at 0, 0.5, 1, 2 and 4 hr after lidocaine or saline administration. Neither analgesics nor sedatives was given during the evaluation period. RESULTS: Lidocaine time-dependently diffused across the tracheostomy tube cuff. Thirty and 60 min after cuff inflation lidocaine concentrations in the water bath reached approximately 8 and 17 microg x ml(-1) representing 160 and 340 microg in 20 ml of water, respectively. The VAS decreased from 53.5 +/- 10.6 to 25.1 +/- 9.8 mm (P < 0.01) 0.5 hr following lidocaine administration which continued until the end of evaluation period. In the placebo group, VAS did not change. CONCLUSION: Lidocaine diffusion across the tracheostomy tube cuff reduces tube discomfort. PMID- 10831197 TI - Head elevation reduces head-rotation associated increased ICP in patients with intracranial tumours. AB - PURPOSE: To quantify the effects of graded head rotation and elevation on intracranial pressure (ICP) in neurosurgical patients, before and after induction of general anesthesia. METHODS: Patients with supratentorial tumours (n=12), scheduled for craniotomy with planned ICP monitoring, underwent baseline ICP measurements awake and supine (0 degrees rotation and elevation). Incremental degrees of head rotation (15 degrees) and of head elevation (10 degrees) were performed independently and in combination. Paired measurements of ICP at all levels of head rotation and elevation were also performed before and after induction of general anesthesia (n=6). RESULTS: The baseline ICP was 12.3 +/- 6.4 mmHg (n=12). Changes of ICP were proportional to the degree of head rotation or elevation. Head rotation of 60 degrees maximally increased ICP to 24.8 +/- 14.3 mmHg (P < 0.05). Head elevation above 20 degrees reduced ICP with a maximal reduction to -0.2 +/- 5.5 mmHg at 40 degrees elevation (P < 0.01). Head elevation to 30 degrees reduced the intracranial hypertension associated with head rotation. No differences were observed between ICP measurements made before or after induction of general anesthesia (n=6). Three patients experienced headache with extreme head rotation (<60 degrees) and intracranial hypertension (ICP > 20 mmHg). CONCLUSION: Head rotation of 60 degrees caused an increase in ICP. Concomitant head elevation to 30 degrees reduced the intracranial hypertension associated with head rotation. Headache with head rotation may provide a useful clinical warning of elevated ICP. PMID- 10831198 TI - Distribution of randomised controlled trials of drugs for post-operative nausea and vomiting. AB - PURPOSE: Randomised controlled trials (RCTs) guide therapeutic decisions. But which RCTs are done; which omitted; and which should be done? This study illustrates a method to explore these questions applied to drugs for post operative nausea and vomiting (PONV). METHODS: Review articles listed 18 drugs for PONV. All RCTs of these drugs for PONV were sought. The first drug mentioned in an RCT was counted and tabulated against others in all the arms of the RCT (against itself in a dose-ranging RCT). Additional drugs mentioned in these RCTs were added to the study, for a total of 40 drugs. RESULTS: Drugs involved in the most RCTs were: ondansetron 131 RCTs; propofol 118; droperidol 74; metoclopramide 67; granisetron 52; scopolamine 22; tropisetron 16. Drugs involved in the fewest RCTs: two drugs with 2 RCTs; twelve drugs with one; three with none. Probability that this distribution occurred by chance: P < 0.00001; that the distribution of dose-ranging RCTs occurred by chance: P < 0.001. Regression of RCT numbers on cost: R = 0.86, P < 0.0001; on year of drug introduction: R = 0.14. Of 1600 possible comparisons of drugs for PONV, (including dose-ranging) 97.8% have never been published. CONCLUSION: Although some antiemetic drugs for PONV have been studied in large numbers of RCTs, many have not been adequately evaluated. Finding relevant RCTs and tabulating their comparison arms is useful for directing future research, and is applicable to any symptom or disorder. PMID- 10831199 TI - Tracheal intubation of outpatients with and without muscle relaxants. AB - PURPOSE: To compare intubating conditions and postoperative myalgias in outpatients after intubation with propofol/alfentanil compared with propofol/alfentanil/succinylcholine with and without precurarisation with d tubocurarine. METHODS: 144 ASA I-II ambulatory patients for dental extraction under anesthesia were studied. Subjects received either 3 mg d-tubocurarine (Group II) or saline (Groups I, III) i.v. prior to induction of anesthesia with 20 microg x kg(-1) alfentanil and 2.5 mg x kg(-1) propofol followed by 1.5 mg x kg(-1) succinylcholine (II and III) or saline 0.9% (I) for muscle relaxation. The ease of airway management and the postoperative incidence, severity and distribution of muscle pains were examined. RESULTS: Intubation was successful in all patients and there were no differences in jaw mobility, ease of bag-mask ventilation, visualization of the vocal cords or cord position. Limb movement was more common during intubation in Group I (37.5%) than in Group III (8.3%) or Group II (2%), P < 0.05. At home, VAS scores for myalgia were higher in Group III than in Group I and II. Neck myalgia was more common in Group II (72%) than in Groups II (44%) and I (41%), P < 0.05. Myalgias were also more common in Group III patients (P < 0.05). CONCLUSION: Acceptable intubating conditions were achieved with propofol and alfentanil alone. Succinylcholine reduced limb movement during intubation but was associated with postoperative myalgias for up to five days. Precurarisation with tubocurarine reduced the severity of succinylcholine myalgia. PMID- 10831200 TI - Cardiovascular responses to anesthetic induction in patients chronically treated with angiotensin-converting enzyme inhibitors. AB - PURPOSE: To investigate the effects of chronic ACE inhibition on cardiac neural function following induction of general anesthesia in patients with underlying coronary artery disease. METHOD: In a prospective case-control study, heart rate variability (HRV) and baroreflex control were compared preoperatively and 30 min after anesthesia induction in patients receiving, or not, ACEI (n=16, control group and n=16, ACEI group). All patients had normal cardiac function and anesthesia consisted of a fixed dose regimen of fentanyl and midazolam. Anesthesia-related hypotension was defined by systolic blood pressure < 90 mmHg. Spectral density of HRV was calculated for low frequency and high frequency bands (LF, from 0.05 to 0.15 Hz and HF, from > 0.15 to 0.6 Hz). Baroreflex sensitivity was estimated after blood pressure changes induced by injections of phenylephrine (PHE) and nitroglycerin (NTG). RESULTS: The HRV parameters and baroreflex sensitivity were not different between groups, during the awake and anesthesia periods. Anesthesia produced similar reduction in total HRV in the Control and ACEI groups (-93 +/- 28% vs -89 +/- 32%), and in baroreflex sensitivity during NTG (-64 +/- 21% vs -54 +/- 17%) or PHE tests (-74 +/- 25% vs -72 +/- 22%). Anesthesia-related hypotension occurred in nine patients in the ACEI group (vs two controls). Although the hypertensive response to phenylephrine was greater after anesthesia in both groups, the sensitivity to phenylephrine was attenuated in those patients experiencing hypotension in the ACEI group. CONCLUSIONS: Chronic preoperative treatment with ACEIs does not influence cardiac autonomic regulation and anesthetic-induced hypotensive episodes are mainly attributed to decreased alpha-adrenergic vasoconstrictive response. PMID- 10831201 TI - Autonomic circulatory and cerebrocortical responses during increasing depth of propofol sedation/hypnosis in humans. AB - PURPOSE: To describe the relative effects of graded central nervous system (CNS) depression, using increasing propofol infusion rates, on neurovegetative brainstem-mediated circulatory control mechanisms and higher cortical activity in healthy humans. METHODS: Propofol was administered using an infusion scheme designed to achieve three target blood concentrations in ten healthy volunteers. Blood propofol concentrations and sedation scores were determined at baseline, during the three propofol infusion levels, and 30 min into the recovery period. Electroencephalographic (EEG) power was measured in three frequency bands to quantify cortical activity, and autonomic heart rate control was quantified using spontaneous baroreflex assessment and power spectral analysis of pulse interval. RESULTS: Sedation scores closely paralleled propofol blood concentrations (0, 0.53 +/- 0.34, 1.24 +/- 0.21, 3.11 +/- 0.80, and 0.96 +/- 0.42 microg x mL(-1) at baseline, three infusion levels and recovery respectively), and all subjects were unconscious at the deepest level. Indices of autonomic heart rate control were decreased only at the deepest levels of CNS depression, while EEG effects were apparent at all propofol infusion rates. These EEG effects were frequency specific, with power in the beta band being affected at light levels of sedation, and alpha and delta power altered at deeper levels. CONCLUSIONS: The results of this study support a relative preservation of neurovegetative circulatory control mechanisms during the early stages of CNS depression using gradually increasing rates of infusion of propofol. Indices of circulatory control did not reliably reflect depth of sedation. PMID- 10831202 TI - [Anesthesia in a child presenting a anhydrotic ectodermic dysplasia associated with a multiminicore myopathy]. AB - PURPOSE: To report the perioperative management of anesthesia and analgesia in a child presenting with the association of multiminicore myopathy (MMM) and anhydrotic ectodermic dysplasia (AED). CLINICAL FEATURES: An eight-year-old girl was admitted for elective orthopedic surgery of the lower limbs. AED is a congenital dermatosis characterized by the absence of sweating and subsequent problems with thermoregulation; in addition, maxillary hypoplasia and abnormal teeth can render intubation difficult. MMM is a rare congenital myopathy characterized by proximal muscle weakness, stable in time or with a slow and progressive evolution. It can involve respiratory muscles and be associated with severe cardiomyopathy. Moreover, MMM shares some characteristics with Central Core Disease which is known to be associated with malignant hyperthermia. Since depolarizing muscle relaxants and halogenated agents could not be used, a combined propofol-based intravenous anesthesia with lumbar epidural analgesia was chosen. This combination provided stable anesthesia, smooth recovery and excellent analgesia during and after the operation, without complications. Temperature was monitored closely during surgery and in the postoperative period. CONCLUSIONS: The association of MMM and AED requires rapid distinction between hyperthermia secondary to anhydrosis and malignant hyperthermia. The management should provide a "trigger-free" anesthetic and optimal postoperative analgesia without sedation. If appropriate for the surgical procedure, a combination of general with regional anesthesia is particularly attractive in achieving these objectives. PMID- 10831203 TI - The use of remifentanil for Cesarean section in a parturient with recurrent aortic coarctation. AB - PURPOSE: To illustrate the clinical utility of a short acting opioid (remifentanil) based general anesthetic for Cesarean section in a parturient with compromised cardiac function. CLINICAL FEATURES: A 23-yr-old primigravida, complicated by a recurrent aortic coarctation with an approximate 50% narrowing of the aortic arch, presented for elective Cesarean section at 37 wk gestational age. Initially asymptomatic, her clinical condition had deteriorated as the pregnancy progressed, with worsening episodes of mild chest pain and shortness of breath. A semi-elective Cesarean section under general anesthesia was planned at 37 wk to minimize the potential for aortic complications associated with the hemodynamic stress of labour. Remifentanil was infused at 0.05 to 0.1 microg x kg(-1) x min(-1) with good sedation and analgesia for the placement of invasive monitors. The infusion was increased to 0.2 microg x kg(-1) x min(-1) for induction, and combined with isoflurane 0.4 to 0.6% for maintenance of anesthesia. The patient maintained stable hemodynamics throughout and her trachea was extubated without difficulty at the end of the procedure. The newborn did not require tracheal intubation, mask ventilation or naloxone and was in excellent condition upon transfer to the well baby nursery. CONCLUSION: Remifentanil, when used as part of an opioid-based general anesthetic for Cesarean section, can provide maternal hemodynamic stability with minimal neonatal respiratory depression and should allow for immediate postoperative tracheal extubation of the mother. PMID- 10831204 TI - Inadvertent placement of pulmonary artery catheter into right carotid artery. AB - PURPOSE: To report a case of misplacement of a pulmonary artery catheter (PAC) into the carotid artery after open heart surgery. CLINICAL FEATURES: A 20-mo-old boy underwent open heart surgery (VSD repair). On the first day postoperatively, he had severe pulmonary hypertension and a PAC was inserted via the left internal jugular approach without complication. Two hours later, chest radiography showed the PAC in the right internal carotid artery which it had reached via the right and left ventricles and aorta. The PAC was withdrawn and a new PAC was inserted and its position was confirmed by chest radiography. Two years later echocardiography failed to demonstrate the second VSD or a residual leak through the patch although a PAC could be passed from the right ventricle to the left ventricle and subsequently into the aorta and right carotid artery. CONCLUSION: Correct placement of a PAC should be confirmed by chest radiography or other techniques to prevent complication. PMID- 10831205 TI - Does intrathecal morphine alter the stress response following coronary artery bypass grafting surgery? AB - PURPOSE: Intrathecal morphine administered prior to coronary artery revascularization (CABG) surgery was studied to determine its effects on the stress response. METHODS: In a single centre, open, randomized clinical trial, first time elective CABG surgery patients, < 75 yr, were studied. Control subjects (n=12) received a standardized anesthetic consisting of fentanyl (maximum cumulative dose of 35 microg x kg(-1)), propofol, and pancuronium. In addition, spinal subjects (n=13) received 1.0 mg (age > 60 yr) or 1.5 mg (age < or = 59 yr) intrathecal morphine prior to induction of anesthesia. Control subjects received continuous i.v. morphine at 2 mg x hr(-1) on arrival in the ICU with i.v. bolus morphine supplementation as required while spinal subjects received bolus i.v. morphine as required. Changes in plasma cortisol and catecholamine concentrations were measured preoperatively, poststernotomy, on admission to ICU, following tracheal extubation, at 0800 hr on the first postoperative day, and 24 and 48 hr after ICU admission. RESULTS: No differences between groups were detected for demographic variables. The percent change in cortisol concentration relative to preoperative values (control vs spinal; (38 (87) vs -41 (46)%: P < 0.05)) was lower in the spinal group on admission to ICU. The percent change in plasma epinephrine levels (control vs spinal) on admission to ICU (285 (337) vs -10 (37)%) and 0800 hr after surgery (314 (341) vs -4 (37)%) was also significantly different. CONCLUSION: Intrathecal morphine only partially attenuated the postsurgical stress response in CABG surgical patients. PMID- 10831206 TI - Effects of intravenous and local anesthetic agents on omega-conotoxin MVII(A) binding to rat cerebrocortex. AB - PURPOSE: The cellular target site(s) for anesthetic action remain controversial. In this study we have examined any interaction of i.v. anesthetics (thiopental, pentobarbital, ketamine, etomidate, propofol, alphaxalone), local anesthetics (lidocaine, prilocaine, procaine and tetracaine), and the non anesthetic barbiturate, barbituric acid with the omega-conotoxin MVII(A) binding site on N type voltage sensitive Ca2+ channels in rat cerebrocortical membranes. METHODS: [125I] omega-conotoxin MVII(A) binding assays were performed in 0.5 ml volumes of Tris.HCl buffer containing BSA 0.1% for 30 min at 20 degrees C using fresh cerebrocortical membranes (5 microg of protein). Non-specific binding was defined in the presence of excess (10(-8) M) omega-conotoxin MVII(A). The interaction of i.v. (alphaxolone, etomidate, propofol, pentobarbitone, ketamine and thiopentone), local (lidocaine, prilocaine, procaine and tetracaine) anesthetics and barbituric acid was determined by displacement of [125I] omega-conotoxin MVII(A) (approximately 1 pM). RESULTS: The binding of [125I] omega-conotoxin was concentration-dependent and saturable with Bmax and Kd of 223 +/- 15 fmol/mg protein and 2.13 +/- 0.14 pM, respectively. Unlabelled omega-conotoxin MVII(A) displaced [125I] omega-conotoxin MVII(A) yielding a pKd of 11.04 +/- 0.04 (9.2 pM). All i.v. and local anesthetics at clinically relevant concentrations did not show any interaction with the omega-conotoxin MVII(A) binding site. CONCLUSION: The present study suggests that omega-conotoxin MVII(A) binding site on N-type voltage sensitive Ca2+ channels may not be a target for i.v. and local anesthetic agents. PMID- 10831207 TI - Detection of subdural placement of epidural catheter using nerve stimulation. AB - PURPOSE: To report the detection of a subdural catheter placement using nerve stimulation through an epidural catheter. CLINICAL FEATURES: An 85-yr-old gentleman was scheduled for radical cystectomy and creation of an ileal conduit. Combined general anesthesia and regional technqiue was selected. An epidural catheter (19 G Arrow Flextip Plus) was inserted prior to induction of general anesthesia. Intra-operatively, the patient received 5 mg morphine and 10 ml bupivacane 0.5% via the epidural catheter. The patient remained hemodynamically stable throughout the operation and did not require intravenous opioids. The patient was discharged to the ward with an order for epidural morphine for pain control. The next day, the patient remained comfortable. As an ongoing quality assessment to survey the success rate of epidural catheters at our institution, all patients are invited to have their catheter assessed using an electrical epidural stimulation test. Electrical stimulation (1-10 mA) with a segmental motor response (truncal or extremities movement) indicates that the catheter is in the epidural space. No motor response indicates that it is not. In this case, subdural catheter placement was suspected because a diffuse motor response including right anterior chest wall, back muscle, and bilateral lower extremities was observed using only 0.3 mA. Subdural catheter placement was subsequently confirmed by a radiograph showing a very thin film of dye spreading cephalad and caudad over many segments. CONCLUSION: This new electrical test helps to detect subdural placement objectively. PMID- 10831209 TI - POGO score. PMID- 10831208 TI - New media. PMID- 10831210 TI - Transesophageal echocardiography in the management of anaphylactic shock. PMID- 10831211 TI - Blind intubation using the Endotrol tube and a light wand. PMID- 10831212 TI - Averaging powers. PMID- 10831213 TI - Bardet-Biedl syndrome. AB - This case report describes the presentation of a patient with Bardet-Biedl syndrome. Bardet-Biedl Syndrome is an autosomal recessive condition that includes retinal dystrophy, dystrophic extremities (commonly polydactyly), obesity, hypogenitalism, and renal disease. Cognitive deficit has also been considered part of the syndrome. The historically associated Laurence-Moon syndrome includes spastic paraparesis but not the obesity and polydactyly. They are now considered separate conditions. The most common feature of Bardet-Biedl syndrome is retinal dystrophy. The appearance of the retina in the condition is quite variable with typical retinitis pigmentosa being present in only a minority of cases. The associated optic atrophy can be primary in nature and might play a role in the decreased central vision. Diagnosis of the condition is important for visual prognosis and low vision management. The renal disease often goes undetected until specific radiological testing is done after diagnosis of Bardet-Biedl syndrome. This is significant in that early death often occurs in this condition because of the renal disease. PMID- 10831214 TI - The repeatability of discrete and continuous anterior segment grading scales. AB - PURPOSE: To investigate the repeatability of three anterior segment clinical grading scales: 1) verbal descriptors scale (VDS), 2) photographic matching scale (PS), and 3) continuous matching scale (CS). METHODS: Five optometrists graded 30 slides each of 3-9-o'clock staining, bulbar redness, and palpebral conjunctival roughness twice, separated by at least a day. VDS and PS were five-point scales (0-4) with half grades permitted. The CS was a 5-second, 240-frame video movie generated using morphing software. PS and CS grading was done with references presented on a computer screen. RESULTS: Averaged across observers, the test retest intraclass correlation, correlation coefficient of concordance, and Pearson's r ranged from 0.95 to 0.99 (all p < 0.001). Coefficients of repeatability using CS to grade all three ocular conditions ranged between 0.31 and 0.49. The corresponding PS and VDS coefficients of repeatability ranged between 0.37 and 0.49; PS generally had better repeatability than VDS. CONCLUSIONS: Each of the clinical grading scales was reliable. The coefficients of repeatability showed that bulbar redness and palpebral conjunctival roughness were graded with higher precision using CS. PMID- 10831215 TI - Overnight orthokeratology. AB - PURPOSE: Orthokeratology is defined as the temporary reduction in myopia by the programmed application of rigid gas-permeable contact lenses. New reverse geometry contact lens designs and materials have led to a renewed interest in this field. The purpose of this study is to assess visual, refractive, topographic, and corneal thickness changes in subjects undergoing overnight orthokeratology. METHODS: Ten myopic subjects (mean age, 25.9+/-3.9 years) were recruited for a 60-day trial of overnight orthokeratology using reverse geometry rigid contact lenses. After commencing lens wear, subjects were examined on days 1, 7, 14, 30, and 60 at several times throughout the day. High- and low-contrast logarithm of the minimum angle of resolution (logMAR) visual acuity, monocular subjective refraction, autorefraction, autokeratometry, corneal topography, corneal thickness, and slit lamp examinations were performed at each session. RESULTS: Eight subjects completed the study. Both high- and low-contrast uncorrected visual acuity improved significantly by day 7. The mean change in uncorrected high contrast visual acuity at day 60 was -0.55+/-0.20 logMAR (mean at day 60, -0.03+/-0.16; Snellen equivalent, 20/19). The mean change in uncorrected low-contrast visual acuity at day 60 was -0.48+/-0.26 logMAR (mean at day 60, +0.22+/-0.23; Snellen equivalent, 20/33). The mean subjective refraction and autorefraction were significantly reduced from baseline at day 60 (mean change in subjective refraction, +1.83+/-1.23 D; mean change in autorefraction, +0.64+/-0.52 D). Corneal topography showed significant central flattening (mean change in apical radius, +0.20+/-0.9 mm; mean change in shape factor, -0.11+/ 0.18 at day 60). The central cornea also showed significant thinning (mean change, -12+/-11 microm at day 60). All visual, refractive, and topographic outcomes were sustained over the course of an 8-h day. CONCLUSIONS: Overnight orthokeratology is an effective means of temporarily reducing myopia. The possible mechanism of corneal remodeling through central corneal thinning is discussed. PMID- 10831216 TI - Distance telescopes: a survey of user success. AB - The distance telescope has a historical reputation for causing difficulties in prescribing and adaptation. Hence, we considered that a retrospective survey of patients at Nottingham Low Vision Clinic might elucidate specific attributes that influence an individual patient's success in using a distance telescope. From 142 patients prescribed distance telescopes since the Clinic's inception, 133 apparently remained users and were mailed a preliminary three-question enquiry about usage of their distance telescopes. The 87 respondents were followed up with questionnaire 2, requesting explicit information about usage, namely frequency, degree of ease or difficulty, and purpose. Older patients required higher magnification (p < 0.025). Seventeen of 74 respondents to questionnaire 2 had various adaptational problems, which are discussed; 57 of 74 patients found their distance telescopes easy to use, and 49 of 57 were frequent users. Thus, ease and frequency are linked (p < 0.05). People tended to use their distance telescopes outdoors and indoors with similar frequency (p > or = 0.29). Adaptation was found to be unrelated to visual acuity, binocularity/monocularity, ocular pathology, or restricted mobility; magnification seemed to be influential, although not significantly. Aging did not significantly impede adaptation. We infer that the universal criterion for selecting treatable patients seems to be personality type. We conclude that adaptation to a device is dependent upon active recognition of its benefits, paralleled with a tolerance of its constraints, which combine to make usage easy and regular on at least one common task. PMID- 10831217 TI - Relative legibility and confusions of letter acuity targets in the peripheral and central retina. AB - BACKGROUND: Patients with macular disease may image optotypes at a peripheral retinal locus during visual acuity testing. In this study, we asked if the relative legibility and confusions between letters are similar in the fovea and at 10 degrees in the peripheral retina. METHODS: Twenty five upper-case alphabet letters (all except "I"), constructed to the same specifications as the Sloan letters, were presented one at a time on a computer monitor at the fovea and at 10 degrees in the temporal, superior and superior-temporal retina. RESULTS: The range of relative legibility at peripheral loci was generally larger than in the fovea, for all letters and for the subset of 10 Sloan letters. Although many confusion pairs were similar in the fovea and periphery, additional confusion pairs, preferentially involving curved letters, occurred uniquely in the periphery. CONCLUSION: The increased range of relative legibility for letter targets in the peripheral retina underscores the importance of using letter-by letter scoring to obtain precise measures of visual acuity in patients without central vision. PMID- 10831218 TI - Synthesis and antimicrobial activity of copper(II) and manganese(II) alpha,omega dicarboxylate complexes. AB - Copper(II) alpha,omega-dicarboxylate complexes of general formulae, [Cu(O2C(CH2)nCO2)].xH2O, [Cu(O2C(CH2)nCO2) (phen)2].xH2O and [Cu(O2C(CH2),CO2)(bipy)y].xH2O (n = 1-8; y = 1, 2; phen = 1,10-phenanthroline; bipy = 2,2'-bipyridine) were synthesised. These copper complexes, some related manganese(II) complexes and the metal-free ligands were screened in vitro for their ability to inhibit the growth of Candida albicans. Metal-free 1,10 phenanthroline and all of the copper(II) and manganese(II) phenanthroline complexes were potent growth inhibitors, with only one bipyridine complex, [Cu(O2C(CH2)CO2)(bipy)2].2H2O, having moderate activity. The remaining substances were effectively inactive. Complexes which were active against C. albicans also proved effective against C. glabratta, C. tropicalis and C. kreusi with the manganese complexes retaining superior activity. For the phenanthroline complexes the active drug species is thought to be the dication [M(phen)2(H2O)n]2+ (M = Cu, Mn). Escherichia coli and Staphylococcus aureus were resistant to all of the metal complexes and also to metal-free 1,10-phenanthroline. Only the copper phenanthroline complexes showed intermediate activity against Pseudomonas aeruginosa. PMID- 10831219 TI - Meeting report--copper research at the top. AB - In this brief paper, the author reports on a meeting on copper research (2nd International Meeting on Copper Homeostasis and its Disorders: Molecular and Cellular Aspects) recently held in Ravello, Italy (17-21 September 1999). Aimed at elucidating the diverse roles played by copper ions in biology and medicine, as they are currently intensely investigated worldwide, the meeting has been organized around a number of major topics from prominent areas of copper research. These included the molecular and cellular basis of copper transport, molecular advances in Menkes and Wilson's diseases, the involvement of copper in neurodegenerative diseases, the structure and function of copper metalloproteins. PMID- 10831220 TI - Metal complexes of lactoferrin and their effect on the intracellular multiplication of Legionella pneumophila. AB - The action of bovine lactoferrin saturated with iron, zinc and manganese on the intracellular multiplication of Legionella pneumophila in HeLa cells has been tested. The results obtained showed that lactoferrin did not influence the invasive efficiency of Legionella. The intracellular multiplication of the bacterium was inhibited by apo-lactoferrin and by lactoferrin saturated with manganese and zinc, whereas lactoferrin saturated with iron enhanced the intracellular growth. Experiments in parallel were performed with iron, manganese and zinc citrate to test the effect due to the metal ions alone. Even in this condition the addition of an iron chelate enhanced the multiplication of Legionella while the manganese chelate produced a certain inhibition. PMID- 10831221 TI - Purification and characterization of a ferredoxin from Haloarcula japonica strain TR-1. AB - A ferredoxin (Fd) was purified from the extremely halophilic archaeon, Haloarcula japonica strain TR-1, to electrophoretic homogeneity. The apparent molecular weight (Mr) of the Fd was estimated to be 24,000 on SDS-polyacrylamide gel electrophoresis. The amino acid composition analysis revealed that the Fd composed of a number of acidic amino acids (uncorrected for amides). The N terminal amino acid sequence (30 residues) was determined to be: PTVEYLNYEVVDDNGWDMYDDDVFAEASDM. The iron content was 3.42+/-0.04 mol/mol-Fd on the basis of the apparent Mr value. The absorption and ESR spectra of the Fd showed similarity to those of Fds from plant and Halobacterium halobium. These results led us to conclude that the H. japonica Fd contained a [2Fe-2S] cluster. PMID- 10831222 TI - Absorption of iron in rats with experimental enteritis. AB - Inflammatory conditions of the gastrointestinal tract and iron-deficiency anemia are very common in humans. Acute intestinal inflammation was pathologically established in rats by intraluminal administration of acetic acid into the duodenum and the proximal jejunum. The study included two control groups of intact (untreated) rats and sham-operated (saline-treated) rats for each intestinal segment. A third group of rats received acetic acid. The acetic acid induced inflammatory process was established histopathologically and biochemically. Two days after treatment, iron absorption was measured using ligated 10-cm loops of proximal jejunum or ligated duodenum in which 59Fe was injected intraluminally (n = 6 in each group). In another four control groups (intact and sham-operated for each intestinal segment) and two acetic acid treated groups, serosal-luminal secretion of 59Fe was measured after intravenous injection (n = 5 in each group). 59Fe transfer from the lumens of the duodenum and jejunum to the portal system was significantly lower in those rats in whom inflammation was induced by acetic acid. There was no apparent serosal-luminal secretion of intravenously injected 59Fe in any of the studied groups. We conclude that acetic acid-induced intestinal inflammation significantly reduces iron absorption by the duodenum and the proximal jejunum. PMID- 10831223 TI - Fusarinines and dimerum acid, mono- and dihydroxamate siderophores from Penicillium chrysogenum, improve iron utilization by strategy I and strategy II plants. AB - Cucumber, as a strategy I plant, and Maize as a strategy II plant, were cultivated in hydroponic culture in the presence of a ferrated siderophore mixture (1 microM) from a culture of Penicillium chrysogenum isolated from soil. The siderophore mixture significantly improved the iron status of these plants as measured by chlorophyll concentration to the same degree as a 100-fold higher FeEDTA supply. Analysis of the siderophore mixture from P. chrysogenum by HPLC and electrospray mass spectrometry revealed that besides the trihydroxamates, coprogen and ferricrocin, large amounts of dimerum acid and fusarinines were present which represent precursor siderophores or breakdown products of coprogen. In order to prove the iron donor properties of dimerum acid and fusarinines for plants, purified coprogen was hydrolyzed with ammonia and the hydrolysis products consisting of dimerum acid and fusarinine were used for iron uptake by cucumber and maize. In short term experiments radioactive iron uptake and translocation rates were determined using ferrioxamine B, coprogen and hydrolysis products of coprogen. While the trihydroxamates revealed negligible or intermediate iron uptake rates by both plant species, the fungal siderophore mixture and the ammoniacal hydrolysis products of coprogen showed high iron uptake, suggesting that dimerum acid and fusarinines are very efficient iron sources for plants. Iron reduction assays using cucumber roots or ascorbic acid also showed that iron bound to hydrolysis products of coprogen was more easily reduced compared to iron bound to trihydroxamates. Ligand exchange studies with epi-hydroxymugineic acid and EDTA showed that iron was easily exchanged between coprogen hydrolysis products and phytosiderophores or EDTA. The results indicate that coprogen hydrolysis products are an excellent source for Fe nutrition of plants. PMID- 10831224 TI - Effects of subchronic alternating cadmium exposure on dopamine turnover and plasma levels of prolactin, GH and ACTH. AB - This study was undertaken to analyze if the effects of subchronic alternating cadmium exposure on pituitary hormone secretion are mediated by changes in dopamine turnover in an age dependent way or are directly correlated to cadmium accumulation at the hypothalamic-pituitary axis. Male rats were treated sc. from day 30 to 60 (prepubertal period) or from day 60 to 90 (adult age) of life, with cadmium chloride (CdCl2) at a dose of 0.5 and 1.0 mg kg(-1) bw, every 4th day in an alternate schedule, starting with the smaller dose. Dopamine (DA) turnover, expressed as the ratio of acid 3.3-dihidroxifenil acetic (DOPAC)/DA in various hypothalamic areas, the plasma levels of prolactin, growth hormone (GH) and adrenocorticotropic hormone (ACTH), and cadmium accumulation in the hypothalamus and pituitary were studied. Prepubertal cadmium exposure decreased DA content in all hypothalamic areas studied, although its turnover was not modified. A decrease in plasma ACTH levels with no changes in plasma prolactin and GH levels were found. Cadmium did not accumulate in pituitary while it increased in the hypothalamus. Metal exposure during adulthood decreased DA content in mediobasal and posterior hypothalamus, and its turnover in posterior hypothalamus and median eminence. It decreased plasma prolactin and ACTH levels but not those of GH. Cadmium concentration increased in both hypothalamus and pituitary. These results suggest that cadmium exposure produces age dependent changes on the secretory mechanisms of the pituitary hormones studied, related to the selective accumulation of the metal at both hypothalamic and hypophyseal level changes. However the effects of the metal are not mediated by dopamine. PMID- 10831225 TI - Characterization of ferritin and ferritin-binding proteins in canine serum. AB - Ferritin and ferritin-binding proteins in canine serum were characterized. A certain percentage of ferritin in canine serum, but no tissue ferritin, was precipitated by centrifugation at 16,000 x g for 30 min. The precipitated ferritin was found to contain two subunits corresponding to the H and L subunits of canine liver ferritin by immunoblotting, the H subunit being predominant. More ferritin was precipitated from canine sera which had been incubated with anti-rat liver ferritin antibody than from untreated sera, and the H chain also predominated. To evaluate the possibility that the autoantibody was responsible for the precipitation of canine serum ferritin, the ferritin-binding activities of canine antibodies were examined using liver ferritin-coated microtiter plates and alkaline phosphatase-labeled antibodies specific for canine IgM, IgA, and IgG heavy chains. The results showed that IgM and IgA, but not IgG, had considerable ferritin-binding activities. Given these results, we suggest that there is H chain-rich isoferritin in canine serum, and that ferritin exists as an immune complex. PMID- 10831226 TI - A gene of the major facilitator superfamily encodes a transporter for enterobactin (Enb1p) in Saccharomyces cerevisiae. AB - While in fungi iron transport via hydroxamate siderophores has been amply proven, iron transport via enterobactin is largely unknown. Enterobactin is a catecholate type siderophore produced by several enterobacterial genera grown in severe iron deprivation. By using the KanMX disruption module in vector pUG6 in a fet3delta background of Saccharomyces cerevisiae we were able to disrupt the gene YOL158c Sce of the major facilitator super family (MFS) which has been previously described as a gene encoding a membrane transporter of unknown function. Contrary to the parental strain, the disruptant was unable to utilize ferric enterobactin in growth promotion tests and in transport assays using 55Fe-enterobactin. All other siderophore transport properties remained unaffected. The results are evidence that in S. cerevisiae the YOL158c Sce gene of the major facilitator super family, now designated ENB1, encodes a transporter protein (Enb1p), which specifically recognizes and transports enterobactin. PMID- 10831227 TI - Kinetics of SO4(-2) reduction under different growth media by sulfate reducing bacteria. AB - Sulfate Reducing Bacteria (SRB) were used to reduce the SO4(-2) concentration in waste water. The growth pattern of SRB was found by varying the concentration of nutrients and the biomass. The specific reaction constant was evaluated in each case. PMID- 10831228 TI - On the ability of four flavonoids, baicilein, luteolin, naringenin, and quercetin, to suppress the Fenton reaction of the iron-ATP complex. AB - Four flavonoids, baicilein, luteolin, naringenin, and quercetin were investigated for their ability to suppress the Fenton reaction characteristic of the iron-ATP complex. Absorption spectroscopy indicates that under the conditions of 18.75% aqueous methanol, 0.0625 mM HEPES pH 7.4 buffer and 1.5:1 quercetin/iron-ATP ratio a mix ligand complex formed. All four flavonoids were found to interfere with the voltammetric catalytic wave associated with the iron-ATP complex in the presence of H2O2. Quercetin and luteolin were able to completely suppress the catalytic wave of the iron-ATP/H2O2 system when a minimum ratio of 1.5:1 of the flavonoid to iron-ATP was reached. At this ratio, the ability of the studied series of flavonoids to suppress the Fenton reaction characteristic of iron-ATP follows as quercetin approximate to luteolin > naringenin approximate to baicilein. Both quercetin and luteolin contain catechol on the B ring, which may enhance the iron chelation of these species over baicilein and naringenin. The common structural feature of all of these flavonoids is the 4-keto, 5-hydroxy region, which may also contribute to the chelation of iron. PMID- 10831229 TI - Microbial pathogens with impaired ability to acquire host iron. AB - Successful microbial pathogens must be adept in obtaining growth-essential iron from healthy hosts. Some potential pathogens, however, are sufficiently impaired in iron acquisition ability so as to be dangerous mainly in hosts with such iron loading conditions as alcoholism, asplenia, hemochromatosis, beta-thalassemia major, or tobacco smoking. The association of six impaired pathogens (Capnocytophaga canimorsis, Yersinia enterocolitica and Y. pseudotuberculosis, Vibrio vulnificus, Tropheryma whippelii, and Legionella pneumophila) with iron loaded humans is described. PMID- 10831230 TI - Iron and citric acid: a fuzzy chemistry of ubiquitous biological relevance. AB - This paper briefly presents a review concerning the species which can arise when iron salts and citric acid are mixed together. The data commented on are required for a correct interpretation of the chemical processes which play a paramount role in biology and in the biological studies involving iron-citrate complexes. PMID- 10831231 TI - Growth hormone secretagogue actions on the pituitary gland: multiple receptors for multiple ligands? AB - 1. Growth hormone (GH) secretion is thought to occur under the reciprocal regulation of two hypothalamic hormones, namely GH-releasing hormone (GHRH) and somatostatin (SRIF), through their engagement with specific cell-surface receptors on the anterior pituitary somatotropes. 2. In addition to GHRH and SRIF, synthetic GH-releasing peptides (GHRP) or GH secretagogue(s) (GHS) regulate GH release through the activation of a novel receptor, the GHS receptor (GHS-R). 3. The cloning of the GHS-R from human, swine and rat identifies a novel G protein-coupled receptor involved in the control of GH secretion and supports the existence of an undiscovered hormone that may activate this receptor. 4. Varieties of intracellular signalling systems are suggested to mediate the action of GHS, which include changes in intracellular free Ca2+ ([Ca2+]i), cAMP, protein kinases A and C, phospholipase C etc. 5. With regard to the use of signalling systems by GHS, especially a new form of GHRP or GHRP-2, a clear species difference has been demonstrated, supporting the possibility of more than one type of GHS-R. PMID- 10831232 TI - Atrial tachyarrhythmias induced by acetylcholine in tilapia (Oreochromis sp.) isolated atria. AB - 1. Effects of the parasympathetic neuromediator acetylcholine (ACh) on atrial tissues vary greatly depending on the species, the type of atrial cells and experimental conditions. The aim of the present study was to investigate, with microelectrode techniques, the arrhythmogenic effects of ACh in tilapia (Oreochromis sp.) isolated atria at room (22-25 degrees C) and high temperature (37 degrees C). 2. Acetylcholine (1-10 micromol/L) shortened action potential duration (APD), depressed action potential plateau and decreased twitch force in tilapia atria, as it did in human atrial fibres. In addition, ACh induced premature responses and re-entrant tachyarrhythmias (TA; frequency range from 7 to 25 Hz) in five of 19 and 14 of 22 tilapia atria tested at room and high temperature, respectively. The higher incidence of ACh-induced TA at 37 degrees C compared with room temperature was statistically significant. 3. The ACh-induced TA consisted of high-frequency and uniform action potentials accompanied by tension oscillation and elevation of diastolic force (flutter). Acetylcholine induced TA could be readily abolished by atropine (1 micromol/L) and prevented by treatment with agents with local anaesthetic properties, such as 0.1 micromol/L tetrodotoxin or 3 micromol/L quinidine. The antagonistic action of quinidine occurred without significant prolongation of APD. 4. The present findings suggest that pharmacological concentrations of the cholinergic muscarinic agonist ACh readily induce TA (mainly atrial flutter) in tilapia atria, presumably via sodium channel-dependent re-entrant excitation. The poikilothermic tilapia appears to be an appropriate animal model for the study of atrial TA. PMID- 10831234 TI - Islet amyloid polypeptide (amylin) modulates chylomicron metabolism in rats. AB - 1. Amylin is a pancreatic peptide that has been shown to be able to induce a state of peripheral insulin resistance. Hyperamylinaemia, which occurs in type 2 diabetes, may be central to a number of metabolic abnormalities present in the diabetic state. Because lipoprotein metabolism is often disturbed in diabetes, we investigated whether amylin was a regulating factor of lipoprotein metabolism in rats; specifically, whether exogenous amylin influences production and clearance of triglyceride (TG)-rich lipoproteins. 2. When amylin was given acutely to rats or by way of infusion, total plasma TG was significantly elevated. Acute doses of amylin decreased fractional clearance rates of TG-rich lipoproteins by 45%. Hydrolysis of lipoproteins by endothelial lipases was not decreased; rather, amylin appeared to reduce hepatic uptake of TG-rich lipoproteins, following conversion to the remnant form. Consistent with the kinetic data in vivo, cell culture studies found that amylin reduced the high-affinity uptake of remnant lipoproteins, probably by inhibiting low-density lipoprotein receptor expression. 3. We have found that amylin can influence the kinetics of TG-rich lipoproteins in vivo and in vitro. Amylin can reduce chylomicron uptake, most probably by regulating lipoprotein receptors either directly, or via modulation of insulin activity. Increased levels of amylin in type 2 diabetes may contribute to the raised concentration of TG-rich remnant lipoproteins present in this disease. PMID- 10831233 TI - Effects of experimental obstructive jaundice on contractile responses of dog isolated blood vessels: role of endothelium and duration of bile duct ligation. AB - 1. We examined the effects of experimental obstructive jaundice caused by bile duct ligation (BDL) on vascular smooth muscle function, as well as the underlying mechanisms involved, by recording responses to noradrenaline (NA), 5 hydroxytryptamine (5-HT) and acetylcholine (ACh) in canine isolated renal arteries and to NA in isolated mesenteric arteries in vitro. All studies were performed 7 days after the onset of BDL in renal arteries and 3, 7 and 15 days after the onset of BDL in mesenteric arteries. 2. The maximum contraction evoked by both NA and 5-HT was significantly attenuated with no change in agonist potency (pD2 value) in renal arteries with endothelium obtained from 7 day BDL dogs when compared with those from sham-operated controls (SO). However, the reduction almost disappeared when the endothelium was removed. In contrast, no change in the responsiveness of renal arteries to KCl could be detected at 7 day BDL. Endothelium-dependent relaxations produced by ACh were significantly increased in renal artery rings from 7 day BDL dogs, but the endothelium independent relaxations produced by papaverine in BDL preparations were not changed when compared with SO controls. 3. At 7 and 15 days after BDL, the Emax values of the mesenteric ring of BDL dogs to NA were significantly lower than that of SO controls, whereas 3 days after surgery there was no significant difference. The pD2 values in arteries obtained from 15 day BDL animals were significantly lower than those obtained from SO control animals. However, no significant changes in pD2 values were seen 3 and 7 days after the onset of BDL. 4. In conclusion, it is suggested that enhanced production and/or release of nitric oxide, mainly of endothelial origin, is associated with reduced vascular responses to contractile agents in experimental obstructive jaundice and that this effect is related to the duration of obstructive jaundice. These results may explain, at least in part, a cause of hypotension that leads to renal failure in patients with obstructive jaundice. PMID- 10831235 TI - Isoproterenol-induced hypertrophy may result in distinct left ventricular changes. AB - 1. The aim of the present study was to analyse the possible lack of uniformity in isoproterenol (ISO)-induced myocardial hypertrophy. 2. Data obtained for isovolumic hearts isolated from 20 rats treated with ISO (0.3 mg/kg over 8 days) were divided into two groups (H1, n = 10; H2, n = 10) according to the volume (mean+/-SD) needed to change left ventricle diastolic pressure from 0 to 40 mmHg (H1, 184+/-30 microL; H2, 108+/-14 microL). Eight control rats (C; 165+/-37 microL) were used for comparison. 3. In addition to ventricular distensibility differences, the groups differed in terms of myocardial mass (mean+/-SEM: H1, 181+/-3 mg > H2, 166+/-3 mg > C, 136+/-3 mg; P < 0.001), of relaxation constant (H2, 43+/-4 msec > H1, 28+/-2 msec; P = 0.0012) and of maximum developed circumferential stress (C, 145+/-9 kdyn/cm2 = H1, 137+/-6 kdyn/cm2 > H2, 110+/-4 kdyn/cm2; P = 0.002). 4. Our results show that ISO-induced myocardial hypertrophy is not homogeneous. Data obtained for H2, taken as a whole and compared with H1 (smaller myocardial mass and impairment of relaxation, elastic stiffness and force generation), suggest that, in some animals, myocardial necrosis and reparative fibrosis may prevail over the stimulus for myocyte growth. The lack of uniformity of ISO-induced myocardial hypertrophy has not been previously reported and may have contributed to the divergence observed in the literature regarding the functional characteristics of the present model. PMID- 10831236 TI - Antihyperglycaemic and anti-oxidant properties of Andrographis paniculata in normal and diabetic rats. AB - 1. Oxidative stress is believed to be a pathogenetic factor in the development of diabetic complications. In the present study, we investigated the ethanolic extract of the aerial parts of Andrographis paniculata for antihyperglycaemic and anti-oxidant effects in normal and streptozotocin-induced type I diabetic rats. 2. Normal and diabetic rats were randomly divided into groups and treated orally by gavage with vehicle (distilled water), metformin (500 mg/kg bodyweight) or the extract (400 mg/kg bodyweight), twice a day for 14 days. 3. At the end of the 14 day period, the extract, like metformin, significantly increased bodyweight (P < 0.01) and reduced fasting serum glucose in diabetic rats (P < 0.001) when compared with vehicle, but had no effect on bodyweight and serum glucose in normal rats. Levels of liver and kidney thiobarbituric acid-reactive substances (TBARS) were significantly increased (P < 0.0001, P < 0.01, respectively), while liver glutathione (GSH) concentrations were significantly decreased (P < 0.005) in vehicle-treated diabetic rats. Liver and kidney TBARS levels were significantly lower (P < 0.0001, P < 0.005, respectively), whereas liver GSH concentrations were significantly higher (P < 0.05) in extract- and metformin treated diabetic rats compared with vehicle-treated diabetic rats. Andrographis paniculata significantly decreased kidney TBARS level (P < 0.005) in normal rats. Hepatic superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) activities were significantly lower in vehicle-treated diabetic rats compared with vehicle-treated normal rats. The extract, as well as metformin, significantly increased the activity of SOD and CAT, but had no significant effect on GSH-Px activity in diabetic rats. The extract and metformin did not produce significant changes in the activity of these anti-oxidant enzymes in normal rats. 4. Our results show that oxidative stress is evident in streptozotocin-diabetic rats and indicate that the ethanolic extract of A. paniculata not only possesses an antihyperglycaemic property, but may also reduce oxidative stress in diabetic rats. PMID- 10831237 TI - Relationship of cigarette smoking with blood pressure, serum lipids and lipoproteins in young Japanese women. AB - 1. The relationship of cigarette smoking with blood pressure and serum lipids and lipoproteins was studied in 1062 young Japanese women aged 20-39 years of age. 2. After adjusting for age, body mass index (BMI), alcohol intake and physical activity scores, the mean systolic and diastolic blood pressures (SBP and DBP, respectively) did not indicate dose-dependent relationships with cigarette smoking. The largest significant mean differences in SBP (3.5%; P < 0.001), DBP (6.2%; P < 0.018), high-density lipoprotein-cholesterol (HDL-C; 13.2%; P < 0.005), ratio of total cholesterol to HDL-C (13.9%; P < 0.022), triglycerides (TG; 24.1%; P < 0.001) and the logarithmic transformation of TG (log TG; 5.6%; P < 0.001) were found between non-smokers and smokers. 3. When age, BMI, alcohol intake and physical activity scores were included in the forward step-wise multiple regression analysis, there were negative relationships found for cigarette smoking and SBP and DBP and positive relationships for cigarette smoking and TG and log TG. 4. Although the results are somewhat variable, the present study shows that cigarette smoking is negatively associated with SBP and DBP and that there is an association between cigarette smoking and serum lipids and lipoproteins and that smoking has an unfavourable effect on these parameters in young Japanese women. PMID- 10831238 TI - Greater blood pressure-lowering effect of the renin inhibitor EMD 58265 than an angiotensin-converting enzyme inhibitor in two-kidney one-clip Goldblatt rabbit. AB - 1. Renin inhibitors may be more advantageous than either angiotensin-converting enzyme (ACE) inhibitors or angiotensin (Ang) antagonists in blocking the renin angiotensin system (RAS) because they do not allow accumulation of either AngI or AngII in plasma. 2. Effects of i.v. administration of two human renin inhibitors (EMD 58265 and U 71038) were compared with the ACE inhibitor enalaprilat on mean blood pressure (BP), renal blood flow (RBF) and plasma AngI and AngII in the anaesthetized two-kidney one-clip Goldblatt rabbit. 3. At doses of 2-2.5 mg/kg, i.v., EMD 58265 and 5-10 mg/kg, i.v., U 71038, both drugs decreased BP approximately 10 mmHg more than enalaprilat (2-4 mg/kg, i.v.) when given either before or after the ACE inhibitor. None of the three agents had any significant effect on RBF in the face of the lowered BP; however, renal vascular resistance was decreased. A higher dose of enalaprilat (10 mg/kg, i.v.) had no further effect on BP than the lower doses but did cause a marked increase in RBF. 4. Both renin inhibitors markedly decreased plasma AngI, but the high basal level of AngII was less consistently and only modestly affected. Enalaprilat, in either the low dose range or at the high dose, was also not effective in significantly decreasing AngII. 5. The results indicate that renin inhibition in the rabbit with a high circulating AngII level is more effective in lowering BP than ACE inhibition. A high dose of the ACE inhibitor may be required to block the intrarenal RAS, which may account for the increase in RBF. PMID- 10831239 TI - Regulation of sodium, calcium and vitamin D metabolism in Dahl rats on a high salt/low-potassium diet: genetic and neural influences. AB - 1. A dietary combination of high salt and low potassium (HSLK) exacerbates hypertension in Dahl salt-sensitive (DS) rats and renders previously normotensive Dahl salt-resistant (DR) rats hypertensive. In both strains, the severity of hypertension correlates with urinary calcium loss. However, the magnitude of excretory calcium losses is significantly greater in DS rats and is potentiated by chemical sympathectomy in both strains. 2. We hypothesized that a defect in vitamin D metabolism may underlie the observed strain-dependent differences in calcium balance. 3. Arterial blood pressure (ABP), water and mineral balance and serum concentrations of 1,25-dihydroxyvitamin D3 (1,25(OH)2 D3) and 25 hydroxyvitamin D3 (25(OH)D3) were measured in intact and chemically sympathectomized (6-hydroxydopamine; 6-OHDA) DS and DR rats after 8 weeks on a HSLK diet. 4. Chronic ingestion of this diet resulted in marked and moderate levels of hypertension in DS and DR rats, respectively. The hypertension was abated and eliminated by 6-OHDA in the DS and DR strains, respectively. Independent of treatment, DS rats had significantly higher urinary excretion of calcium and reduced intestinal absorption of the ion compared with DR rats. The DS rats had significantly higher serum levels of 1,25(OH)2 D3 and markedly lower serum levels of 25(OH)D3 than DR rats. Chemical sympathectomy tended to increase 1,25(OH)2 D3 and to decrease 25(OH)D3 levels in both strains. 5. These data indicate a genetic difference in vitamin D metabolism between DS and DR rats. The abnormally elevated levels of 1,25(OH)2 D3 in DS rats may be an appropriate compensatory response to excessive excretory calcium loss and reduced target organ sensitivity to the hormone and may, maladaptively, directly contribute to hypertension, by stimulating vascular smooth muscle contractility. PMID- 10831240 TI - Stress-induced elevation of the ST segment in the rat electrocardiogram is normalized by an adrenoceptor blocker. AB - 1. Emotional stress is one of the aetiologies of cardiac accidents, but the precise mechanisms remain to be elucidated. In the present study we used the immobilization stress model in rats to investigate electrocardiographic (ECG) changes. 2. A rapid elevation of the ST segment at the II, III, aVF and precordial leads in the ECG was found and this returned to prestress basal levels after removal of stress. 3. This ECG change was normalized by combined blockade of alpha- and beta-adrenoceptors, but not by administration of a potent coronary vasodilator, such as a calcium channel blocker or nitroglycerin. 4. These results suggest that direct activation of cardiac adrenoceptors by stress may result in angina-like ECG changes. PMID- 10831241 TI - Effects of the Na+/H+ exchange inhibitor cariporide (HOE 642) on cardiac function and cardiomyocyte cell death in rat ischaemic-reperfused heart. AB - 1. Na+/H+ exchange has been implicated in the mechanism of reperfusion injury. We examined the effects of the cardiac-specific Na+/H+ exchange inhibitor cariporide (HOE 642) on postischaemic recovery of cardiac function and cardiomyocyte cell death (i.e. necrosis and apoptosis). 2. Rat isolated and buffer-perfused hearts were subjected to 25 min normothermic global ischaemia followed by 120 min reperfusion. Cariporide (10 micromol/L) or its vehicle (0.01% dimethylsulphoxide) was administered for 15 min before ischaemia and for the first 30 min after reperfusion. 3. Cariporide significantly improved the recovery of isovolumic left ventricular function (heart rate, left ventricular developed pressure and left ventricular end-diastolic pressure) and coronary flow throughout reperfusion. Creatine kinase release during reperfusion was significantly less in the cariporide-treated heart. In situ terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL)-positive cardiomyocytes were also significantly less in the cariporide-treated heart after 120 min reperfusion. Electron microscopy showed necrotic changes without typical apoptotic features in cardiomyocytes after reperfusion. Such necrotic changes were mitigated by cariporide. Simultaneous detection of necrotic and apoptotic cardiomyocytes using propidium iodide (PI) and Annexin V revealed that cardiomyocytes in the infarct area were stained with only PI or both PI and Annexin V. Cariporide did not alter the pattern of cardiomyocyte staining with PI and Annexin V, although the number of cardiomyocytes stained with PI or PI plus Annexin V was less than that in vehicle-treated hearts. 4. These results suggest that apoptosis is not a major manifestation of cardiomyocyte cell death in the ischaemic-reperfused myocardium and a cariporide-sensitive mechanism of reperfusion injury promotes both necrotic and apoptotic processes of cell death. PMID- 10831242 TI - Tonic potentiation and attenuation produced by membrane depolarization in guinea pig trachealis. AB - 1. We studied how membrane depolarization directly affected intracellular Ca2+ signalling when voltage-operated Ca2+ channels (VOCC) were not available in guinea-pig tracheal smooth muscle. To block VOCC, we used 3 micromol/L verapamil, which completely abolished high K+ (20-60 mmol/L)-induced contraction, and elevation of fura-2 signal. 2. Muscle tone was generated by adding Ca2+ to the extracellular Ca2+-free solution containing prostaglandin (PG)E2 (100 nmol/L) after abolishing basal tone with indomethacin (1 micromol/L). 3. In the absence of verapamil, high K+ (20-60 mmol/L) solution potentiated 2.4 mmol/l Ca2+-induced sustained contractions. Even in the presence of 3 micromol/L verapamil, replacement with 20 and 40 mmol/L K+ solution induced tonic potentiation, which was changed to attenuation with a higher K+ solution (60 mmol/L), lower extracellular Ca2+ concentration ([Ca2+]o) and pretreatment with cyclopiazonic acid (10 micromol/L), a Ca2+ sequestration inhibitor. 4. These results indicate that the balance between depolarization-dependent Ca2+ release and receptor operated cation channel inhibition may determine whether tonic potentiation or attenuation is manifested, depending on the availability of VOCC, the magnitude of the depolarization, [Ca2+]o and Ca2+ content in the sarcoplasmic reticulum. PMID- 10831243 TI - Functional evidence for anti-oxidant action of fluvastatin on low-density lipoprotein using isolated macrophages and aorta. AB - 1. Fluvastatin has been reported to have not only a hypocholesterolaemic effect, but also a protective effect on low-density lipoprotein (LDL) from oxidation. We functionally evaluated the anti-oxidant effect of fluvastatin on oxidation of LDL by copper ions in vitro using mouse macrophages and rabbit aorta preparations. 2. After native LDL (N-LDL) from rabbit plasma had been pre-incubated in the presence or absence of fluvastatin (10 micromol/L) for 4 h, the N-LDL was mildly oxidized by incubation with 5 micromol/L CuCl2 for 5 h and two oxidized LDL, fluvastatin-pretreated (Flu-OxLDL) and -non-treated (OxLDL), were prepared. The level of thiobarbituric acid-reactive substances (TBARS) in Flu-OxLDL and OxLDL markedly increased compared with N-LDL. The degree of increment was significantly less in Flu-OxLDL than OxLDL. 3. When macrophages were incubated with Flu-OxLDL or OxLDL, the amount of cholesteryl ester that accumulated in the macrophages markedly increased compared with N-LDL. The degree of increment was significantly less in Flu-OxLDL than OxLDL. 4. Acetylcholine-induced endothelium-dependent relaxations in rabbit aortic rings were impaired in the presence of either Flu OxLDL or OxLDL. The degree of impairment was significantly less in Flu-OxLDL. 5. The increased TBARS level, facilitated cholesteryl ester accumulation in macrophages and impaired endothelium-dependent relaxation elicited by OxLDL were not affected by simultaneous treatment with fluvastatin (10 micromol/L). 6. These findings indicate that fluvastatin can protect plasma LDL from oxidative modification and, thereby, prevent cholesterol accumulation in macrophages and endothelial dysfunction in blood vessels. This additional anti-oxidative effect of fluvastatin may be beneficial for preventing the progression of atherosclerosis. PMID- 10831244 TI - Effects of prostaglandins on baroreflex during reperfusion of the ischaemic myocardium. AB - 1. The present study was planned to: (i) determine whether the baroreflex control of heart rate (HR) and renal sympathetic nerve activity (RSNA) was attenuated during reperfusion of short-term ischaemic myocardium; and (ii) study whether blockade of prostaglandin synthesis with indomethacin reversed the inhibitory baroreflex. 2. Arterial pressure was lowered with intravenous sodium nitroprusside before coronary occlusion and 3 min after release of a 5 min occlusion of the left circumflex coronary artery in anaesthetized rabbits. The protocol was repeated 20 min after indomethacin (5 mg/kg, i.v.) or indomethacin vehicle (50 mmol/L tris(hydroxymethyl)aminomethane buffer, pH 8.4) treatment. In addition, this study was performed in a group of vagotomized rabbits. 3. Before indomethacin treatment, the slope of the mean arterial pressure (MAP)-RSNA relationship decreased from -3.3+/-0.77 to -2.01+/-0.69% change in RSNA/mmHg (P < 0.05) during reperfusion of ischaemic myocardium in intact rabbits. The decrease in the slope was reversed by administration of indomethacin. However, the decrease in the slope was not reversed by indomethacin vehicle. Furthermore, the reduction in the slope of the MAP-RSNA relationship during reperfusion of ischaemic myocardium was abolished in vagotomized rabbits. However, there was no inhibition of the slope of the MAP-HR relationship during reperfusion of ischaemic myocardium in either intact or vagotomized rabbits. 4. In conclusion, our data suggest that prostaglandins released by ischaemic myocardium can attenuate the baroreflex-mediated response of RSNA to lowered arterial pressure via vagal afferents during reperfusion of short-term ischaemic myocardium. PMID- 10831245 TI - Beneficial effects of fish oil on glucose metabolism in spontaneously hypertensive rats. AB - 1. Increased interest in fish oil led us to examine their metabolic effects in spontaneously hypertensive rats, which have been reported to have glucose intolerance. 2. Rats were divided into three groups: (i) a control group fed standard rat laboratory chow; (ii) a lard group fed a high-fat diet containing 20% lard; and (iii) a fish oil group fed a high-fat diet containing 20% fish oil for 14 weeks. 3. Systolic blood pressure and fasting blood glucose were markedly increased in the lard group, whereas in the fish oil group they were only transiently increased at the beginning and decreased to levels seen in the control group. 4. Intraperitoneal glucose tolerance test demonstrated that fish oil reversed the impairment of glucose disposal found in the lard group. However, plasma insulin levels were raised transiently at 30 min in the fish oil group compared with the control group. 5. Insulin secretion from pancreatic islets stimulated with glucose in vitro was also enhanced by fish oil. 6. These results lead us to conclude that fish oil improves glucose tolerance by enhancing insulin secretion from pancreatic beta-cells. PMID- 10831246 TI - Antihypertensive effect of cattle bone collagen-derived peptides in ovariectomized stroke-prone spontaneously hypertensive rats. AB - 1. The effect of food collagen, cattle bone collagen-derived (CBC) peptides, on ovariectomy induced increases in blood pressure was examined in stroke-prone spontaneously hypertensive rats (SHRSP). 2. Long-term administration of CBC peptides to ovariectomized SHRSP suppressed the hypertension compared with ovariectomized SHRSP fed standard chow. 3. The CBC peptides showed an inhibitory activity (IC50 = 40 microg/mL) for angiotensin I-converting enzyme (ACE) in vitro. Furthermore, pre-incubation of CBC peptides with gastrointestinal proteases did not change this inhibitory activity of CBC for ACE. 4. These results indicate that CBC peptides may prevent increases in blood pressure in ovariectomized SHRSP by a possible mechanism of an inhibitory action against ACE. PMID- 10831247 TI - Circumventricular organs: definition and role in the regulation of endocrine and autonomic function. AB - 1. The circumventricular organs (CVO) are structures that permit polypeptide hypothalamic hormones to leave the brain without disrupting the blood-brain barrier (BBB) and permit substances that do not cross the BBB to trigger changes in brain function. 2. In mammals, CVO include only the median eminence and adjacent neurohypophysis, organum vasculosum lamina terminalis, subfornical organ and the area postrema. 3. The CVO are characterized by their small size, high permeability and fenestrated capillaries. The subcommissural organ is not highly permeable and does not have fenestrated capillaries, but new evidence indicates that it may be involved in the hypertension produced by aldosterone acting on the brain. 4. Feedback control of corticotropin-releasing hormone (CRH) secretion is exerted by free steroids diffusing into the brain, but substances such as cytokines and angiotensin II act on CVO to produce increases in CRH secretion. Gonadal steroids also diffuse into the brain to regulate gonadotrophin-releasing hormone secretion. Thyrotropin-releasing hormone secretion is regulated by thyroid hormones transported across cerebral capillaries. However, CVO may be involved in the negative feedback control of growth hormone and prolactin secretion. PMID- 10831248 TI - Angiotensin II modulation of the arterial baroreflex: role of the area postrema. AB - 1. Resetting the operating point of the arterial baroreflex appears to be an important factor involved in determining the level of sympathetic outflow. 2. Substantial data indicate that circulating angiotensin (Ang)II can reset the arterial baroreflex to higher operating pressures. 3. This action of AngII to increase the level of sympathetic nervous system outflow relative to blood pressure may contribute to elevated mean arterial pressure (MAP) during AngII hypertension, as well as to the maintenance of MAP during low sodium states. 4. In most instances, the resetting observed during elevated peripheral AngII is dependent on the area postrema (AP). 5. Although the central mechanisms by which the arterial baroreflex resets to different operating pressures remain to be completely defined, the AP may provide a separate mechanism whereby humoral factors can modulate the operating point of the arterial baroreflex. PMID- 10831249 TI - Area postrema and sympathetic nervous system effects of vasopressin and angiotensin II. AB - 1. Precise control over the cardiovascular system requires the integration of both neural and humoral signals related to blood volume and blood pressure. Humoral signals interact with neural systems, modulating their control over the efferent mechanisms that ultimately determine the level of pressure and volume. 2. Peptide hormones such as angiotensin (Ang)II and arginine vasopressin (AVP) act through circumventricular organs (CVO) to influence cardiovascular regulation. 3. The area postrema (AP), a CVO in the brainstem, mediates at least some of the central actions of these peptides. Vasopressin appears to act in the AP to cause sympathoinhibition and a shift in baroreflex control of the sympathetic nervous system (SNS) to lower pressures. These effects of AVP and the AP appear to be mediated by alpha2-adrenoceptor and glutamatergic mechanisms in the nucleus tractus solitarius. 4. In contrast to AVP AngII has effects in the AP to blunt baroreflex control of heart rate and cause sympathoexcitation. The effects of chronic AngII to increase activity of the SNS may be due to AP dependent activation of neurons in the rostral ventrolateral medulla. PMID- 10831250 TI - Brain versus peripheral angiotensin II receptors in hypovolaemia: behavioural and cardiovascular implications. AB - 1. Angiotensin (Ang)II is involved in responses to hypovolaemia, such as sodium appetite and increase in blood pressure. Target areas subserving these responses for AngII include the cardiovascular system in the periphery and the circumventricular organs in the brain. 2. Conflicting data have been reported for the role of systemic versus brain AngII in the mediation of sodium appetite. 3. The role for systemic AngII and systemic AngII receptors in the control of blood pressure in hypovolaemia is well established. In contrast with systemic injections, i.c.v injections of AngII non-peptide AT1 and AT2 receptor antagonists, such as losartan and PD123319, do not reduce arterial pressure in sodium-depleted (furosemide injection plus removal of ambient sodium for 24 h) rats. Thus, brain AngII receptors are likely not important for cardiovascular responses to hypovolaemia induced by sodium depletion. 4. Intracerebroventricular injections of losartan or PD123319 increase arterial pressure when injected at relatively high doses. This hypertensive effect is unlikely to be an agonist effect on brain AngII receptors. Increases in arterial pressure produced by i.c.v. losartan are attenuated by lesions of the tissue surrounding the anterior third ventricle (AV3V). The hypertensive effect of i.c.v. AngII is abolished by lesions of the AV3V. 5. Hypertension induced by AngII receptor antagonists is consistent with hypotension induced by AngII acting in the brain. However, the full physiological significance of this hypotensive effect mediated by brain AngII receptors remains to be determined. PMID- 10831251 TI - Angiotensin and osmoreceptor inputs to the area postrema: role in long-term control of fluid homeostasis and arterial pressure. AB - 1. The role of the area postrema (AP) in the long-term control of body fluid homeostasis and arterial pressure under conditions of increased dietary salt intake is reviewed. A model is proposed in which sympathetic nerve activity is suppressed when dietary salt is increased. It is hypothesized that the AP acts as an essential integrative site in the hind-brain for this response. 2. An essential component of the hypothesis is that basal levels of circulating angiotensin II support arterial pressure in animals consuming a normal salt diet by acting on the AP to drive sympathetic nerve activity. This hypothesis is supported by the observation that the long-term hypotensive response to losartan, the AT1 receptor antagonist, is attenuated in AP-lesioned (APx) rats. 3. The role of hepatoportal sodium receptors in signalling the AP about changes in dietary salt intake is discussed. Intragastric hypertonic saline infusion increases portal venous, but not systemic plasma, osmolality and increases Fos-like immunoreactivity in the AP, nucleus tractus solitarius and the supraoptic, paraventricular and lateral parabrachial nuclei. Other studies have shown that stimulation of these receptors decreases renal sympathetic nerve activity. 4. The hypothesis that the AP is critical in long-term control of arterial pressure and body fluid homeostasis under conditions of altered dietary salt intake was studied. The responses of arterial pressure and sodium and water balance to changes in dietary salt intake were measured in intact and APx rats. Contrary to the hypothesis, APx rats did not exhibit impaired regulation of arterial pressure or water balance. However, APx rats did demonstrate an impaired ability to excrete sodium when salt intake was elevated. 5. Based on these observations, it is concluded that the AP is important in the control of sodium balance, but not arterial pressure, when dietary salt intake is altered. PMID- 10831252 TI - Centrally produced nitric oxide and the regulation of body fluid and blood pressure homeostases. AB - 1. Nitric oxide (NO) tonically inhibits the basal release of vasopressin and oxytocin into plasma. 2. Nitric oxide inhibition on vasopressin secretion is removed, while that on oxytocin is enhanced, during water deprivation, hypovolaemia, moderate osmotic stimulation and angiotensin (Ang)II. This results in a preferential release of vasopressin over oxytocin that promotes conservation of water. 3. Nitric oxide facilitates drinking behaviour stimulated by water deprivation, osmotic stimulation, haemorrhage and AngII. Together with the hormonal response, NO produces a positive water balance during reductions in intracellular and intravascular volumes. 4. Nitric oxide produced within the central nervous system maintains resting arterial blood pressure partially by attenuating the pressor actions of AngII and prostaglandins. 5. Central production of NO is enhanced during osmotic stimulation to counterbalance the salt-induced pressor response. 6. Paradoxically, central production of NO is also enhanced during haemorrhage, presumably to maintain peripheral vasodilation and blood flow to vital organs. PMID- 10831254 TI - Villous tumors of the duodenum: biologic characters and clinical implications. PMID- 10831253 TI - Anteroventral third ventricle periventricular tissue contributes to cardiac baroreflex responses. AB - 1. The studies reviewed in the present paper demonstrate that the anteroventral third ventricle (AV3V) region contains tissue that can modify cardiac baroreflex sensitivity in response to circulating angiotensin (Ang)II and hyperosmolality. 2. The response to hyperosmolality appears to be mediated by noradrenergic receptors. Although the role of noradrenergic receptors in the AV3V region in modification of baroreflex-induced responses to AngII has not been directly tested, this neurotransmitter is a good candidate for control of heart rate because noradrenaline in the AV3V region is critical for mediating other responses to AngII. 3. Results from studies indicate that the AV3V region is part of a central nervous system circuit involved in modulation of cardiac baroreflex sensitivity by circulating substances, possibly acting at the organum vasculosum lamina terminalis. 4. The findings extend the role of the AV3V periventricular tissue as a central site integrating autonomic nervous system function by demonstrating that this brain area contributes to cardiac function, in addition to its well-characterized role in sympathetic nervous system regulation of blood pressure and mechanisms of fluid and electrolyte regulation. PMID- 10831255 TI - Antireflux surgery in Sweden, 1987-1997: a decade of change. AB - BACKGROUND: The aim of this study was to analyse whether new therapeutic options- the introduction of proton-pump inhibitors (PPI) in 1989 and the laparoscopic technique in 1992--altered the surgical treatment of gastro-oesophageal reflux disease (GORD) in Sweden. METHODS: Data obtained from the Centre for Epidemiology (EpC) on patients undergoing surgery for GORD from 1987 to 1997 was analysed, and the information was validated with a questionnaire to all surgical departments. RESULTS: The questionnaire gave a response rate of 94%, and the figures corresponded well with those obtained from the EpC. In 1987, 456 antireflux procedures were performed. Ten years later this figure had increased to 1303. This approximately threefold increase started before the introduction of PPI and was even more pronounced during the following few years. The development of laparoscopic antireflux surgery did not alter this increase. In 1997, 76% of the procedures were performed laparoscopically. The fundoplication rate rose from 5.5 to 12.7 procedures/100,000 inhabitants. The rates varied greatly among different counties; 7 of 23 still had a fundoplication rate of less than 10 in 1997. The median number of procedures per hospital in 1997 was 10. Only two departments accomplished more than 50 antireflux procedures. CONCLUSION: Within 5 years the laparoscopic technique replaced the open procedure as the method of choice. However, the increase in the frequency of antireflux surgery was apparent even before the introduction of laparoscopy. PMID- 10831256 TI - A new method for the detection of incomplete lower esophageal sphincter relaxation in patients with achalasia. AB - BACKGROUND: In patients with early stages of achalasia manometry is of significant diagnostic value. Technically, however, measurement of lower esophageal sphincter (LES) relaxation is not always easy. Accordingly, we looked for a simpler way of measuring incomplete LES relaxation. METHODS: In 186 consecutive patients referred to esophageal motility testing the esophageal body base-line pressure was measured during continuous swilling of 180 ml fluid within 20 sec. RESULTS: Seventeen of the 186 patients had achalasia. Fourteen of these patients were compliant for the swill test, and all had a positive test, characterized by a steady increase in base-line pressure with negative deflections on deglutition. All nonachalasia patients could complete the test, which was negative in all except one patient, who had a severe peptic stricture. CONCLUSIONS: The swill test is diagnostic for incomplete lower esophageal relaxation in achalasia in compliant patients without organic stenosis. PMID- 10831257 TI - Diagnosis of Helicobacter pylori infection in children: is the 15N urine test more reliable than the 13C breath test? AB - BACKGROUND: The [13C]urea breath test is a convenient method to estimate Helicobacter pylori colonization non-invasively. As an alternative method, the [15N]urea urine test has been established. It is hypothesized that the urine test could be advantageous under some circumstances. The diagnostic value of the breath test might, for example, be jeopardized owing to fluctuating 13C isotope dilution by endogenous carbon dioxide. METHODS: To compare the reliability and practicability of the two tests, 13C breath tests and 15N urine tests were performed simultaneously in three groups of patients: A) 36 adults under standardized conditions, B) 67 children who were allowed to move around during the test, and C) 18 children once without and once during standardized physical activity. RESULTS: In the 36 adults there was a close correlation of the test results of the two methods (r = 0.88). In contrast, the correlation in the group of 67 children and infants was poor (r = 0.10). In the children with controlled activity the 13C results fluctuated, whereas the 15N results remained unchanged. CONCLUSIONS: Owing to their identical biochemical basis the breath test and the urine test produce the same diagnostic results in the same individuals if standardized pre-test and test conditions are followed. However, factors such as physical activity during the test may provoke fluctuations with a tendency to decreasing 13C values. This could be due to additional isotope dilution by increases in the endogenous production of 12C-carbon dioxide. We conclude that in very active children the 15N urine test might be advantageous and more reliable than the 13C breath test. PMID- 10831258 TI - Phenotypic and genotypic characterization of Helicobacter pylori from patients with and without peptic ulcer disease. AB - BACKGROUND: Helicobacter pylori plays an important role in peptic ulcer disease, although not all H. pylori-infected persons will develop a peptic ulcer. Currently, H. pylori strains cannot be divided into commensals and pathogens. METHODS: Fifty H. pylori strains were cultured from patients divided into five groups on the basis of upper endoscopic findings: gastric ulcer, duodenal ulcer, gastritis, esophagitis, or normal. The ultrastructural adherence pattern in vivo, autoagglutination, hemagglutination, adhesion to human gastric adenocarcinoma (AGS) cells, and the lipopolysaccharide (LPS) profile of H. pylori strains were recorded; randomly amplified polymorphic DNA (RAPD) and urease gene typing were performed and correlated with diagnostic groups. RESULTS: Electron micrographs showed that H. pylori strains from patients with gastric ulcers adhered more frequently through filamentous strands and were less frequently found free in mucus than any other diagnostic group (P < 0.0001). Neither median hemagglutination titer nor median adhesion capacity to a human gastric adenocarcinoma cell line was related to endoscopic findings. Nevertheless, H. pylori strains from patients with gastric ulcers were more prone to autoagglutinate than were strains from the other diagnostic groups (P = 0.03). H. pylori strains from gastric ulcer patients were found to be more homogeneous, as determined by RAPD and urease gene typing, than strains from the other diagnostic groups (P < 0.01). In addition, a positive correlation was found between a patient's age and the adhesion to AGS cells of the patient's H. pylori strain (P = 0.006). CONCLUSION: A combination of an H. pylori autoagglutination test, RAPD, and urease gene typing may be useful in separating gastric ulcer-related strains from duodenal ulcer-related and non-ulcer dyspepsia-related strains. PMID- 10831259 TI - Effects of caffeine on gastrointestinal myoelectric activity and colonic spike activity in dogs. AB - BACKGROUND: Caffeine is known to produce various effects on the gastrointestinal tract. Few studies have, however, investigated the effects of caffeine on gastrointestinal myoelectric activity under physiologic conditions. The aims of this study were to investigate the effects of caffeine on colonic motility (spike activity) and gastric and jejunal slow waves in conscious dogs. METHODS: The study was performed in eight healthy female hound dogs (15-20 kg) in which one pair of electrodes was implanted on the serosa of the stomach, one pair on the proximal jejunum, and another pair on the ascending colon. The protocol consisted of a 60-min recording as base line and a 90-min recording after intravenous injection of saline, a low dose of caffeine (125 mg), or a high dose of caffeine (250 mg). RESULTS: In the colon there was a significant increase in the number of spike bursts per minute after the injection of both the low (6.69+/-1.27 versus 8.22+/-1.31, P < 0.05) and the high dose (4.38+/-1.23 versus 6.92+/-0.98, P < 0.05) of caffeine. The increase in the total energy of spikes per minute after the injection of the high dose of caffeine was significantly higher than that after the low dose of caffeine (41.2+/-14.6 versus 133.4+/-39.2, P < 0.05). The characteristics of the gastric and jejunal slow waves were, however, not affected by caffeine. CONCLUSION: Caffeine increases spike activity in the colon, and its effect may be dose-dependent. Gastric and jejunal slow waves are not affected by caffeine. PMID- 10831260 TI - Gastric emptying: comparison of scintigraphic, polyethylene glycol dilution, and paracetamol tracer assessment techniques. AB - BACKGROUND: The method of choice for studying gastric emptying is dependent on several factors. The aim of the present study was to assess the concordance between solid gastric emptying, using a scintigraphic technique as gold standard, and gastric emptying as measured with parcetamol tracer and polyethylene glycol (PEG) dilution. METHODS: Two groups of seven male volunteers with similar ages and weights were studied, one for scintigraphic (310-kcal omelette with 12-15 MBq 99mTc-labeled macroaggregated albumin) and paracetamol (1.5 g dissolved in water and administered concomitantly with the omelette) and one for the marker dilution study (PEG 4000 dissolved in a 310-kcal meal). RESULTS: The gastric half-emptying time (T50) was shorter in the PEG study than in the scintigraphic test (47.5 (37.5-62) versus 68.1 (43.6-89.4), median (range) (P < 0.05), respectively), whereas there was no significant difference between the T50 for gastric emptying as assessed with the scintigraphic and paracetamol tracer methods. No difference in gastric emptying rate using PEG dilution or paracetamol tracer was obtained. CONCLUSIONS: In summary, this study shows that scintigraphic, paracetamol tracer, and PEG dilution methods can all be used to assess gastric emptying. The use of the paracetamol tracer technique offers a relatively inexpensive technique that yields a good approximation of gastric emptying as verified by the scintigraphic emptying of a solid meal. PMID- 10831261 TI - Effects of neural blocking agents on motor activity and secretion in the proximal and distal rat colon: evidence of marked segmental differences in nicotinic receptor activity. AB - BACKGROUND: Neuromodulation may be a new therapeutic approach in inflammatory bowel disease, but very little is known about neural control of colonic secretion in vivo. We therefore determined the effects of neural blockade on colonic motor activity and mucosal secretion in anaesthetized rats. METHODS: A proximal and a distal colonic segment were isolated in four groups of chloralose-anaesthetized rats (n = 8 in each group), and we measured luminal pressure and transmural potential difference (PD) as a marker of electrogenic chloride secretion. Recordings were made from proximal and distal segments simultaneously, which made it possible to directly compare response patterns. RESULTS: Under control conditions luminal pressure waves were associated with phasic, lumen-negative increases in PD which had a significantly greater magnitude and longer duration in the distal colon. Atropine blocked both pressure waves and PD waves in the proximal colon, but some PD waves, although of lower magnitude, remained in the distal colon. Hexamethonium abolished pressure waves in both segments and induced a marked reduction in PD in the distal but not in the proximal colon. Lidocaine also reduced PD, more so in the distal colon, and dissociated the pressure-PD linkage. CONCLUSION: In the distal but not in the proximal colon, there is a strong nicotinic, neurogenic 'tone' that maintains a high basal secretory activity. The results encourage the search for neuromodulatory agents in the treatment of colonic secretory disease. PMID- 10831263 TI - Prevalence of Coeliac disease in the general population of northern Spain. Strategies of serologic screening. AB - BACKGROUND: Coeliac disease (CD) is common in Western countries. In Spain, however, no studies exist on its prevalence. The best method for serologic screening is also unknown. METHODS: We determined the IgA and IgG antigliadin antibodies (AGA) and IgA antiendomysium antibodies (AEA) in a sample of 1170 randomly selected subjects from a health area in northern Spain. The prevalence of CD was calculated with a two-step serologic screening method (first IgA and IgG AGA and then AEA in those positive for AGA) or with only one step with AEA determination. Diagnosis was confirmed by small-intestinal biopsy. RESULTS: When using a two-step strategy, we found 174 (15%) subjects with AGA. Only one of these was confirmed by means of AEA (prevalence of CD = 0.9/1000). Two subjects were found to be positive when a one-step strategy was used (prevalence of CD = 1.7/1000). This yielded an economic saving of 19% with regard to the use of a two step strategy. One new case of CD in a seronegative subject was diagnosed during the study (real prevalence of CD = 2.6/1000; 95% confidence interval = 0.7-8.2). CONCLUSIONS: The prevalence of CD in Northern Spain is 2.6/1000 (1:389). One-step serologic screening (AEA) is both more economical and more sensitive than the two step procedure (first AGA and then AEA if AGA is positive). PMID- 10831262 TI - Bacterial translocation in experimental shock is dependent on the strains in the intestinal flora. AB - BACKGROUND: Enteric microorganisms are responsible for a significant proportion of post-surgical infections. Intestinal mucosal injury may permit translocation of bacteria and endotoxin. This study investigates translocation in peritonitis and ischemia/reperfusion by inoculating different bacterial species into the small intestine. METHODS: Twenty-five pigs were monitored hemodynamically and divided into three groups: controls (C), ischemia/reperfusion (I/R), and peritonitis (P). Intramucosal pH (pHi) was calculated tonometrically. A perfusion tube was positioned in the ileum for inoculation of the bacterial strains. In a first study period a non-pathogenic bacterium was used, whereas Escherichia coli strains with known ability to translocate were used in a second. Blood and mesenteric lymph nodes (MLNs) were obtained for bacterial culture and endotoxin analyses. RESULTS: Mesenteric arterial blood flow and pHi decreased in groups I/R and P. Endotoxin levels increased in these groups in period 1, whereas in period 2 an increase over time was only observed in group P. No bacterial translocation to blood or MLNs occurred in period 1. In period 2 bacteria translocated to MLNs in all animals, including controls. Translocation to central and/or mesenteric venous blood was found in all groups, but mainly in I/R and P. The incidence of mucosal injury was similar in the two periods. CONCLUSIONS: Since positive blood and MLN samples were only found in period 2, we conclude that translocation of bacteria seems to be more dependent on the presence of translocating strains in the intestinal bacterial flora than on the mucosal insult. PMID- 10831264 TI - Virus serology in familial Crohn disease. AB - BACKGROUND: Recent findings that early-in-life virus infections represent a risk factor for Crohn disease, that exacerbations of disease sometimes appear associated with common viral infections, and, in particular, suggestions that Crohn disease may be the result of persistent infection with measles virus prompted serologic studies for antibody to 19 common viruses, Chlamydia psittaci, and Mycoplasma pneumoniae. METHODS: Sera from 14 affected members of 2 French families with a high frequency of Crohn disease and from age- and sex-matched controls, taken in 1990 and 1992, and from unaffected family members were tested. Complement fixation, enzyme immunoassay, and indirect immunofluorescent tests were used. RESULTS: There were no significant differences between patients and controls or between affected and unaffected family members of family 2, with the exception that affected siblings carried higher antibody titers for mycoplasma one year and for varicella another year. There were no differences in measles IgM and IgG or mumps IgG antibody levels between patients and controls, or between patients and unaffected family members. CONCLUSIONS: Serology failed to find evidence of participation by 19 common viruses, C. psittaci, and M. pneumoniae in Crohn disease. The data do not support the hypothesis that persistent measles virus infection causes Crohn disease. PMID- 10831265 TI - Intestinal mucosal secretion of basic fibroblast growth factor in patients with ulcerative colitis. AB - BACKGROUND: Basic fibroblast growth factor (bFGF) promotes angiogenesis and several other biologic processes, including proliferation of mesenchymal cells and tumor progression. We investigated whether bFGF could be detected in the intraluminal secretion of the small intestine, sigmoid colon, and rectum in healthy individuals and in patients with ulcerative colitis. METHODS: We used endoscopic perfusion techniques to obtain samples from well-defined intestinal segments. The perfusion fluid concentrations of bFGF, biochemical markers of inflammation, myeloperoxidase (MPO), interleukin-6 (IL-6), and permeability (albumin) were determined with immunochemical methods. RESULTS: In the perfusion fluids the albumin concentration, which reflects passive diffusion, was less than 1% of the plasma concentration, whereas the intestinal concentration of bFGF was similar to that in plasma. Among healthy subjects the concentration of bFGF was eightfold higher in the jejunum and twofold higher in the rectum than in the sigmoid colon. The perfusion fluid from colorectal segments in patients with ulcerative colitis had a significantly higher mean concentration of bFGF than that from healthy individuals; an almost 10-fold difference was found in rectal segments. There were strong correlations between the concentration of bFGF and the concentrations of MPO and IL-6. CONCLUSIONS: The high concentrations of bFGF in the intestinal perfusion fluid reflect either a local synthesis or an active secretion of bFGF within the mucosa. The bFGF concentration differs in intestinal anatomic location and increases significantly in patients with ulcerative colitis in close relationship with biochemical markers of inflammation and permeability. PMID- 10831266 TI - Tissue concentration of platelet-derived endothelial cell growth factor in colorectal cancer. AB - BACKGROUND: Angiogenesis is essential for the continuous growth of tumour cells under unfavourable conditions in patients. Experimentally, platelet-derived endothelial cell growth factor (PD-ECGF) promotes tumour proliferation by stimulating angiogenesis. However, the clinical significance and regulating mechanism of its production in colorectal cancer are not well understood. METHODS: The tissue concentration of PD-ECGF in colorectal neoplasm and normal mucosa was determined. The systemic oxygenation and nutritional status of the patients were also evaluated. RESULTS: The mean concentration of PD-ECGF in the cancer was significantly higher than that in the normal mucosa or adenoma. The tissue concentration of PD-ECGF in the cancer was associated with the clinicopathologic findings, including the tumour size, serosal invasion, lymphatic vessel involvement, and lymph node metastasis. It was also correlated with the patient's age, levels of PO2 and O2 saturation in arterial blood, and the variables reflecting nutritional status. The multivariate regression model showed that the serum concentration of cholinesterase, the arterial level of PO2, lymph node metastasis, and the tumour size were the independent factors that influenced the tissue concentration of PD-ECGF in colorectal cancer. In contrast, these factors were not associated with the PD-ECGF concentration in normal mucosa. CONCLUSIONS: PD-ECGF may play an important role in the progression of colorectal cancer. Systemic deterioration of oxygenation and nutritional condition in wasted patients may also lead to local activation of PD-ECGF specifically in the cancer tissue. PD-ECGF may be indispensable for maintaining relentless growth of colorectal cancer, and the control of its expression may be of therapeutic importance. PMID- 10831267 TI - Efficacy of octreotide and sclerotherapy in the treatment of acute variceal bleeding in cirrhotic patients. A prospective, multicentric, and randomized clinical trial. AB - BACKGROUND: Sclerotherapy is the most widely used method for treatment of acute variceal bleeding. Previous reports have suggested that octreotide infusion is as effective as sclerotherapy. Our aim was to investigate the efficacy and safety of octreotide in comparison with sclerotherapy in controlling variceal bleeding. METHODS: Seventy-six cirrhotic patients were randomized to receive either sclerotherapy (n = 37) or octreotide (n = 39) infusion of 50 microg/h intravenously for 48 h after a bolus of 100 microg, followed by subcutaneous injection of 100 microg/8 h for an additional 72 h. RESULTS: The two groups were similar in base-line data. A similar initial control of bleeding was obtained in 94.6% for sclerotherapy and 84.6% for octreotide (NS). No difference was observed between sclerotherapy and octreotide in rebleeding (23% versus 33%) and treatment failure (22% versus 36%, respectively). Furthermore, the overall success of treatment was 78% for sclerotherapy and 64% for octreotide. No significant difference in mortality was observed between treatments (eight patients for octreotide and three patients for sclerotherapy, NS). CONCLUSIONS: These results show that both treatments present a very high and similar initial and final control of bleeding. However, there is a trend that could be clinically important towards better results in the patients treated with sclerotherapy. PMID- 10831268 TI - Effects of prostacyclin inhibition on splanchnic hyposensitivity to glypressin in a hemorrhage-transfused rat model of portal hypertension. AB - BACKGROUND: Hyposensitivity to vasopressin is a well-documented phenomenon in animals with portal hypertension and patients with cirrhosis and hemorrhage. Similar findings exist with infusion of glypressin (a long-acting vasopressin analogue), and this phenomenon could be ameliorated by inhibition of nitric oxide (NO) synthase. Besides NO, excessive formation of prostacyclin (PGI2) has been shown to play an important role in the development of hyperdynamic circulation and the mediation of hyporeactivity to vasoconstrictors in portal-hypertensive states. This study was designed to investigate whether the blockade of PGI2 activity by indomethacin infusion could enhance the portal-hypotensive effect of glypressin in portal-hypertensive rats with bleeding. METHODS: Portal hypertension was induced by partial portal vein ligation (PVL). Fourteen days after operation systemic and portal hemodynamics were measured in stable or bleeding PVL rats receiving intravenous glypressin (0.07 mg/kg) or indomethacin (5 mg/kg) followed by glypressin infusion. In rats with a hypotensive hemorrhage 4.5 ml of blood was withdrawn, and 50% of the withdrawn blood was reinfused before the administration of glypressin or indomethacin. RESULTS: Splanchnic hyposensitivity to glypressin was shown in hemorrhage-transfused PVL rats. Indomethacin infusion did not cause significant systemic and portal-hemodynamic changes in bleeding PVL rats (P > 0.05). The addition of indomethacin significantly enhanced the portal-hypotensive effects of glypressin and potentiated the increases in mean arterial pressure induced by glypressin infusion in bleeding PVL rats. CONCLUSIONS: The improvement of splanchnic hyposensitivity to glypressin in a hemorrhage-transfused rat model of portal hypertension by the administration of indomethacin suggests that PGI2 has in the development of this hyposensitivity. PMID- 10831269 TI - Effects of 1-year interferon-alpha 2a treatment in patients with chronic hepatitis C and persistently normal transaminase activity. AB - BACKGROUND: Certain chronic hepatitis C carriers have persistently normal transaminase activity. The aims of this study were to determine the virologic and histologic effects of 1 year of interferon-alpha treatment in such patients. METHODS: Thirty-one patients were followed up in our Liver Unit. Eleven accepted interferon-alpha therapy; the 20 others were not treated and served as controls. Interferon-alpha, 3 MU, was given thrice weekly for 1 year. Serum was examined for hepatitis C virus (HCV)-RNA before, at the end of, and 6 months after treatment. Liver biopsy was performed 6 months after the cessation of treatment in 10 of 11 treated patients (one refused biopsy) and after a mean of 30.6+/-22.7 months in the 20 untreated patients. RESULTS: At the end of follow-up two of the treated patients had undetectable serum HCV-RNA and five had increased alanine aminotransferase (ALAT) values. In contrast, only one of the untreated patients had abnormal ALAT activity. All 20 untreated patients were constantly viremic. No significant histologic improvement was observed in the treated patients evaluated by means of post-treatment liver biopsy. The mean annual progression rate of fibrosis was very slow and similar in the treated and untreated patients (0.09 (range, 0-0.62) versus 0.07 (range, 0-0.60) fibrosis units). CONCLUSIONS: One year of interferon-alpha treatment can suppress HCV-RNA in patients with chronic hepatitis C and persistently normal ALAT values followed up over long periods. The rate of fibrosis progression in such patients is very slow, and therapeutic strategies should take this fact into account. Antiviral treatment is debated for patients without fibrosis in initial biopsy specimens. PMID- 10831270 TI - Objective assessment of the contribution of each diagnostic test and of the ordering sequence in jaundice caused by pancreatobiliary carcinoma. AB - BACKGROUND: Computer-assisted diagnostic systems are not substantially more accurate than the clinician in the differential diagnosis of jaundice but may help in optimal selection and sequencing of tests. The present study aimed to assess with an electronic diagnostic tool the pattern of ordering tests and the diagnostic contribution and related financial cost of each test in jaundiced patients with pancreatobiliary carcinoma, in an effort to make the clinician's diagnostic behaviour more efficient and economical. METHODS: Clinical and diagnostic test data were prospectively gathered from 356 jaundiced patients with pancreatobiliary carcinoma and entered in a Bayesian diagnostic programme. The test results were added to the existing diagnostic evidence, and the programme calculated the diagnostic contribution of each test. RESULTS: A total of 1804 diagnostic tests were ordered. Quantitative assessment of the diagnostic contribution of each test showed that percutaneous transhepatic cholangiography and computed tomography were associated with the highest provision of information. The most cost-effective tests were ultrasonography and liver function tests. CONCLUSIONS: It is possible to determine objectively the diagnostic contribution of each test in establishing the diagnosis of pancreatobiliary carcinoma. The observed physician behaviour in ordering the various diagnostic tests might be improved with regard to its efficacy and its cost-effectiveness profile. PMID- 10831271 TI - Severe pancreatitis after parathyroidectomy. AB - Cope showed in 1957 that pancreatitis may be the presenting symptom in hyperparathyroidism. Since then, the literature has reported a coincidence of primary hyperparathyroidism and pancreatitis between 1% and 19%, but the true relationship has not been fully established. When severe pancreatitis follows parathyroidectomy, a condition familiar to parathyroid surgeons, reports are mostly anecdotal and by many authors considered to be coincidental. We present the case history of a 58-year-old man with a longstanding history of untreated primary hyperparathyroidism who developed severe pancreatitis immediately after removal of a 400-mg parathyroid adenoma. He was the first in a series of 108 operated patients to develop this complication. His preoperative levels of parathormone and serum calcium were the highest in our material. We believe that pancreatitis after parathyroidectomy is a real but rare complication that might be predicted by preoperative high values of serum calcium and parathormone. PMID- 10831272 TI - Combined chemoimmunotherapy in metastatic melanoma--is there a need for the double? AB - The therapeutic benefit of adding immunotherapeutic agents such as interferon (IFN)-alpha and interleukin (IL)-2 to established single-agent or combination chemotherapy regimens for the treatment of metastatic melanoma has not been proven. On the contrary, recent studies indicate a significantly higher incidence of severe toxic side effects in patients treated with combined biochemotherapy. This article summarizes recent trials using either chemotherapy alone or chemotherapy plus the administration of IFN-alpha and/or IL-2 to evaluate the benefit of a combined biochemotherapy. PMID- 10831273 TI - Monotherapy with docetaxel in second- or third-line treatment of anthracycline resistant metastatic breast cancer. AB - Nineteen breast cancer patients pretreated with one or two anthracycline containing regimens for visceral metastases received i.v. docetaxel 100 mg/m2 on day 1, q 21d. Docetaxel was administered as second-line therapy in 11 patients, whereas eight patients received docetaxel in a third-line setting. In the second line setting, complete response (CR) was achieved in two (18%), partial response (PR) in four (36%) and stable disease (SD) in three (27%) patients resulting in a response rate (RR) of 54%. In the third-line setting three (38%) patients experienced PR (RR 38%) and two (25%) SD. In the second-line setting, median time to progression was 6.5+/-3.9 months (range 2.1-15.8) versus 4.7+/-5.5 months (range 0.6-15.9) in the third-line setting. Median overall survival was 9.6+/-8.0 months (range 2.7-25.8) versus 11.2-6.1 months (range 4.8-18.7). Overall, no patient experienced treatment-limiting toxicities. We conclude that docetaxel induced responses in 48% of anthracycline-resistant patients enrolled into the present study. The safety profile of docetaxel was manageable and tolerable. Docetaxel represented efficacious treatment in patients with metastatic breast cancer progressing despite previous anthracycline-containing chemotherapy. PMID- 10831274 TI - Preoperative UFT administration for patients with advanced colorectal cancer- increased uptake of 5-fluorouracil by tumor tissue is a prognostic factor. AB - The aim of this study was to clarify whether increased 5-fluorouracil (5-FU) uptake by tumor tissue following preoperative UFT administration is a prognostic factor after surgery in colorectal cancer patients. We examined the concentrations of 5-FU in tumor or normal tissue of 96 colorectal cancer patients who received UFT (400 mg/day) orally for 7 days prior to surgery. Patients were divided into two groups with high or low 5-FU concentrations in tumor tissue (defined as higher or lower than the cut-off value, respectively). The cut-off value of 5-FU was established based on the upper limit of the 95% confidence interval of the median of the concentration found in normal tissue (0.106 microg/g). Of the 96 patients, 62 (64.6%) were in the low-5-FU group and 34 (35.4%) in the high-5-FU group. The latter had a more favorable clinical outcome (p=0.0465). Cox regression analysis revealed that two independent variables, stage and 5-FU status in tumor tissue, were significant for prediction of survival. These findings suggest that increased uptake of 5-FU by tumor tissue following preoperative oral administration of UFT is an independent prognostic factor in colorectal cancer patients. This variable needs to be considered in the design of future therapeutic trials. PMID- 10831275 TI - Induction of cytokines and killer cell activities by cisplatin and 5-fluorouracil in head and neck cancer patients. AB - It has been suggested that certain antitumor agents stimulate antitumor immunity. In the present study, we examined whether cisplatin and 5-fluorouracil (5-FU) accelerate the antitumor host responses in head and neck cancer patients. Two groups of patients were studied, i.e. an untreated (UT) group and a treated, disease-free (TDF) group that received chemo-immunotherapy in combination with radiotherapy and operation. When peripheral blood mononuclear cells (PBMC) derived from head and neck cancer patients were treated with cisplatin or with 5 FU, interferon-gamma, tumor necrosis factor (TNF)-alpha, TNF-beta, interleukin (IL)-1beta, IL-6, IL-12 and IL-18 as well as killer cell activities were significantly induced in both groups. In this case, these activities induced by cisplatin in UT showed lower levels than those in TDF, whereas the activities induced by 5-FU in the UT group demonstrated almost similar levels to those in TDF. These activities were significantly inhibited by anti-asialo-GM1 antibody. Furthermore, cytokine levels in sera and killer activities of PBMC derived from the cancer patients were significantly increased after cisplatin administration. These findings suggest that cisplatin and 5-FU increase anticancer immunity mediated by induction of cytokines and killer cell activities in patients with head and neck cancer. PMID- 10831276 TI - Vinorelbine and interferon-alpha2c as second-line therapy in metastatic renal cell carcinoma. AB - Second-line treatment of patients with metastatic renal cell carcinoma (MRCC) progressing under therapy with biological response modifiers (BRM) is an unresolved issue. Thirty-seven patients with MRCC progressing under treatment with BRM received vinorelbine i.v. at a dose of 30 mg/m2 q 22 days and 4,800,000 IU interferon (IFN)-alpha2c s.c. thrice weekly. Partial remission (PR) occurred in 8% of patients, stable disease (SD) (median duration 8, range 3-35+ months) was observed in 46% of patients. Median overall survival was 15 (range 1-49) months. No major toxicities occurred. Patients with MRCC who failed first-line treatment with BRM had a high chance to enter PR or SD under combined, low-toxic therapy with vinorelbine and IFN-alpha2c. PMID- 10831277 TI - Inhibition of Ras p21 synthesis by antisense undecamers with uniform and specifically arranged phosphorothioate linkages. AB - The design of chimeric oligodeoxynucleotides (ODNs) in which certain phosphodiester linkages are replaced by phosphorothioate (PS) aims to decrease non-sequence-specific effects of uniform PS ODNs and to preserve the PS-provided protection against exo- and endonucleases. This study has, for the fist time, directly compared the differences in nuclease resistance, cellular uptake, antisense potency and sequence specificity of PS and end-capped, pyrimidine protected (PPS) undecamer ODNs, that are complementary to the initiation codon region of human Ha-ras mRNA. At concentrations above 5 microM, both PS and PPS undecamers were moderately and equally stable for over 48 h in complete medium with RS485 cells overexpressing Ha-ras. They were completely stable at 0.4 microM when complexed with Lipofectin reagent that enhanced cellular uptake up to 9 fold. Both the antisense PPS and PS undecamers produced well-defined inhibition of Ras p21 synthesis in both cell-free and cell-based assays. However, non sequence-specific effects of the uniform phosphorothioates were still significant. In contrast, the antisense PPS undecamer, when delivered to RS485 cells with Lipofectin reagent, inhibits human Ras p21 synthesis by more than 90% at a concentration of 3.2 microM, while the effect of controls with inverted, mismatched or scrambled sequence was minimal (5% or less) on p21 synthesis and RS485 cell growth. PMID- 10831278 TI - Low-dose twice-daily fractionated X-irradiation of ovarian tumor cells in vitro generates drug-resistant cells overexpressing two multidrug resistance-associated proteins, P-glycoprotein and MRP1. AB - Failure of chemotherapy is frequently observed in patients previously treated with radiotherapy. To establish a cellular model for examining this resistance phenotype a series of mammalian tumor cell lines were exposed in vitro to fractionated X-irradiation and were then shown to express resistance to multiple antitumor drugs, including vincristine, etoposide and cisplatin. In these experiments the radiation was delivered as 10 fractions of 5 Gy (dose resulting in 1 log cell kill) given intermittently over several months. We now report that a comparable multidrug-resistance profile is expressed by human SK-OV-3 human ovarian tumor cells exposed in vitro to low dose (2 Gy) twice-daily fractions of X-rays given for 5 days on two consecutive weeks, essentially mimicking clinical practice, involving an overexpression of two MDR-associated proteins, P glycoprotein and the multidrug resistance protein 1 (MRP1), with the latter being readily detectable by immunocytochemistry. PMID- 10831279 TI - Toxicity of anticancer agents mediated by electroporation in vitro. AB - Electroporation is a physical event that temporarily reduces cell membrane barrier properties. Diminished membrane barrier properties are achieved by exposing cells to pulsed electric fields. When a cell has been treated with electric fields it is possible for extracellular agents to gain access to the cell interior. This process has been used in vivo to increase the uptake of chemotherapeutic agents by tumor cells which results in dramatically higher response rates than when drug is used alone. This type of treatment is called electrochemotherapy (ECT); bleomycin is most often used as the drug for this type of treatment. It was hypothesized that electroporation could be used to augment the cytotoxicity of other anticancer agents. Therefore, this study was performed in order to screen 44 different combinations of drug and cell type in vitro to identify drugs that may have higher cytotoxicity when combined with electroporation. Results from seven cell types indicate that the IC50 of bleomycin can be reduced by a factor of 100-5000 when electroporation is used to facilitate internalization. The IC50 values of cisplatin and carboplatin could be reduced by factors ranging from 3 to 13 in six different cell lines as a result of electroporation. These IC50 reductions in multiple cell lines suggest that cisplatin and carboplatin may be effective in vivo as part of ECT treatment. PMID- 10831280 TI - Structure-activity studies on gossypol in tumor cell lines. AB - Gossypol [(2,2'-binaphthalene)-8,8'-dicarboxaldehyde-1,1',6,6',7,7'-hexahydroxy 5,5'-diisopropyl-3,3'-dimethyl] 1a is a naturally occurring compound extracted from the cotton plant and has been extensively studied as an oral male contraceptive. Its favorable toxicity profile, and the more recent demonstration of anti-tumor activity in animals and humans, prompted us to investigate the role of the aldehyde groups in a structure-activity study in cultured tumor cells. Four racemic compounds were evaluated: gossypol 1a, gossypolone 2, the bis Schiff's base of L-phenylalanine methyl ester with gossypol (bis Schiff's base) 1c and apogossypol 1b. The former two compounds both retain the aldehyde functional groups at positions 8 and 8' of the molecule whilst in the latter two compounds the aldehydes are blocked or absent, respectively. In addition, the l- and d-isomers of gossypol 1a, the bis Schiff's base 1c and the half Schiff's base 1d (one aldehyde blocked) were tested. The cell lines studied included melanoma (SK-mel-19), cervix (Sihas), small cell lung (H69) and myelogenous leukemia (K562). Cytotoxicity was measured using the MTT and flow cytometric viability assays. Racemic gossypol 1a and gossypolone 2 induced similar dose-dependent decreases in cell viability in all the cell lines with IC50 values of 23-46 and 28-50 microM, respectively. In contrast, the racemic bis Schiff's base derivative of gossypol 1c and apogossypol 1b showed minimal activity in any cell line up to 50 microM. The l-enantiomer of gossypol 1a was significantly more active than the d-enantiomer (IC50 of 20 versus > 50 microM, respectively). When one aldehyde of either enantiomer was blocked 1d cytoxicity was comparable to the l-enantiomer of gossypol. The data suggest that only one aldehyde group is required for the cytotoxicity of gossypol 1a, irrespective of the stereoconfiguration. PMID- 10831282 TI - Clinical issues and treatment advances in thrombocytopenia: introduction. PMID- 10831281 TI - Efficacy of MGI 114 (HMAF) against the MRP+ metastatic MV522 lung carcinoma xenograft. AB - This study is part of an effort to evaluate efficacy of the novel agent MGI 114 (HMAF) against tumors resistant to conventional chemotherapeutic agents. MGI 114 is a novel semisynthetic anticancer agent currently in chemotherapeutic phase II trials to evaluate activity against various solid tumors. Previous studies indicate MGI 114 was active against human MDR1/gp170+ solid tumor xenografts. Recent evidence suggests overexpression of the MRP protein may also be clinically relevant to development of drug resistance in solid tumors. We evaluated the efficacy of MGI 114 against a human MRP+ lung carcinoma xenograft. Parent MV522 lung carcinoma cells were transfected with a MRP cDNA expression vector and resistant cells selected by exposure to vinblastine (30-fold resistance). Analysis of resistant clones indicated 20- to 40-fold increases in expression of both MRP mRNA and MRP protein. Administration of MGI 114 at the maximum tolerated dose (7 mg/kg, 5 x/week for 3 weeks) to MRP tumor-bearing mice demonstrated this novel agent was active against MRP+ tumors and significantly extended their lifespan (p<0.001). In contrast, other cytotoxic agents had minimal activity against this MRP+ xenograft. These results indicate MGI 114 should retain activity in vivo against MRP+ tumor types. The development of this MRP+ xenograft model, in conjunction with the parent MV522 and MDR1/gp170+ xenograft models, will be useful for screening new classes of agents for activity against multidrug resistant tumors. PMID- 10831283 TI - Current issues with platelet transfusion in patients with cancer. AB - For the past 30 years, platelet transfusions have been used in the treatment of thrombocytopenia caused by decreased production, inadequate function, or increased destruction of platelets. The number of platelet transfusions has increased more than transfusions of other blood components, shifting from whole blood use for the platelet source to plateletpheresis. Hematology/oncology patients are among the largest group receiving platelet transfusions, primarily because the more aggressive chemotherapies produce more acute and prolonged thrombocytopenia. While platelet transfusions often rescue patients with very low platelet levels, they are associated with the risk of viral and bacterial infections, as well as alloimmunization. Platelet donor recruitment can also be difficult, and platelet transfusion can be very expensive depending on the source of platelets. As a result, prophylactic transfusions are less likely to be administered at higher platelet counts, reducing platelet use and cost of platelet transfusions. However, cancer patients receiving intensive chemotherapy or myeloablative regimens require multiple platelet transfusions. For these patients, alternate strategies are needed so that platelet transfusions can be significantly reduced or eliminated. PMID- 10831284 TI - Pharmacologic treatment options in patients with thrombocytopenia. AB - Thrombocytopenia that results from chemotherapy has become an increasingly important issue in the treatment of cancer and remains a difficult clinical problem. The identification of a safe and effective platelet growth factor could significantly improve the management of severe chemotherapy-induced thrombocytopenia. Over the past decade, a number of hematopoietic growth factors with thrombopoietic activity have been identified, including stem-cell factor (c kit ligand), interleukin (IL)-1, IL-3, IL-6, and IL-11, as well as thrombopoietin (TPO) and its derivatives. Only a few of these agents have shown acceptable tolerability and sufficient ability to stimulate thrombopoiesis to justify testing in randomized clinical trials. Currently, IL-11 is the only cytokine licensed in the United States for treatment of chemotherapy-induced thrombocytopenia. However, its thrombopoietic activity is modest and its use is often associated with unfavorable side effects. Identification of TPO, the c-Mpl ligand, as the primary physiologic regulator of megakaryocyte and platelet development offers important promise for treatment of thrombocytopenia. Preliminary clinical studies of recombinant human TPO (rhTPO), a full-length glycosylated molecule, indicate that it is safe and biologically active in reducing severe chemotherapy-induced thrombocytopenia. In addition to rhTPO, the future may see the development of novel genetically engineered, high-affinity cytokine receptor agonists and c-Mpl ligand mimetic peptides. PMID- 10831285 TI - Biologic and structural differences of thrombopoietic growth factors. AB - The search for a thrombopoietic agent has resulted in the identification of numerous cytokines and growth factors with thrombopoietic activity. However, with the exception of interleukin (IL)-11 and thrombopoietin (TPO), the megakaryopoietic activity of most of these molecules has not produced clearly identifiable clinical benefits. Despite the relatively modest effect of IL-11 on megakaryocyte and platelet production in vitro and in vivo, it does reduce the need for platelet transfusions in specialized clinical settings. In contrast, the c-Mpl ligand TPO has been shown to be a potent stimulator of megakaryocyte and platelet production both in vitro and in vivo. Clinical studies are being conducted with two different preparations of the c-Mpl ligand: recombinant human thrombopoietin (rhTPO) and pegylated recombinant human megakaryocyte growth and development factor (PEG-rHuMGDF). A recombinant form of the complete human molecule, rhTPO is glycosylated and produced in mammalian cells. PEG-rHuMGDF consists of only the receptor-binding domain linked to a polyethylene glycol (PEG) moiety and is generated in Escherichia coil. Although c-Mpl ligands are still being evaluated, preliminary evidence indicates that these molecules can elevate platelet counts and may be useful in a range of clinical contexts. This report discusses aspects of the biology behind the clinical actions of IL-11 and the c-Mpl ligands. PMID- 10831286 TI - Clinical experience with recombinant human thrombopoietin in chemotherapy-induced thrombocytopenia. AB - Since the identification and cloning of c-Mpl ligand, two forms of recombinant human thrombopoietin have undergone clinical development. Both the full-length molecule, known as rhTPO, and the truncated version of the molecule, known as pegylated recombinant human megakaryocyte growth and development factor (PEG rHuMGDF), have been evaluated in phase I/II clinical trials in cancer patients receiving myelosuppressive chemotherapy. Early clinical trials with PEG-rHuMGDF in cancer patients demonstrated its clinical safety and platelet-stimulating activity. However, the development of neutralizing antibodies and clinically significant thrombocytopenia in some patients and normal donors who received PEG rHuMGDF have led to discontinuation of clinical trials with this molecule in the United States. Clinical experience with rhTPO so far indicates that this full length glycosylated molecule is remarkably well tolerated and has a favorable safety profile. In these studies, rhTPO exhibited dose-dependent increases in circulating platelet counts and bone marrow megakaryocytes before chemotherapy. In addition, there was an increase in the frequency and proliferation of bone marrow progenitor cells and mobilization of progenitors into the peripheral blood. Early results also showed that rhTPO can attenuate chemotherapy-induced severe thrombocytopenia and reduce the need for platelet transfusions. However, in this setting, the optimal schedule of rhTPO administration may depend on the length of the regimen and anticipated timing of the platelet nadir. These initial results indicate that rhTPO is a safe and potentially useful agent in the prevention and management of chemotherapy-induced thrombocytopenia. Results of larger randomized clinical trials will determine the therapeutic potential of this novel growth factor in various clinical settings. PMID- 10831287 TI - Thrombopoietin in the treatment of acute myeloid leukemia and in stem-cell transplantation. AB - Recent studies indicate that thrombopoietin (TPO) may be highly effective in mobilizing autologous peripheral blood stem cells (PBSCs) for transplantation in patients undergoing intensive chemotherapy. The yield of CD34+ progenitor cells can be increased as can the percentage of patients achieving adequate grafts for use in transplantation. However, the effect of TPO in patients with hematologic malignancies undergoing induction or postremission chemotherapy or in the stem cell transplantation setting has not been demonstrated. Further study is warranted for better definition of the role of TPO in the treatment of severe thrombocytopenia in these settings. PMID- 10831288 TI - Future directions with platelet growth factors. AB - Since the purification of thrombopoietin 6 years ago, c-Mpl ligands such as recombinant human thrombopoietin (rhTPO) and pegylated recombinant human megakaryocyte growth and development factor (PEG-rHuMGDF) have undergone extensive clinical investigation. Both recombinant forms have been shown to reduce the thrombocytopenia associated with nonmyeloablative chemotherapy. Several areas of research have been identified for further clinical development of c-Mpl ligands. One future direction is to continue to explore the intravenous route of administration of rhTPO and PEG-rHuMGDF, as well as fusion proteins of interleukin-3-thrombopoietin and thrombopoietin peptide mimetics, which may be as potent as thrombopoietin, but may lack antigenicity. Another focus would be on the use of these molecules in treating non-chemotherapy-induced thrombocytopenia associated with myelodysplastic syndrome (MDS), idiopathic thrombocytopenic purpura (ITP), human immunodeficiency virus (HIV)-related ITP, and liver disease. Additionally, c-Mpl ligands may have a role in improving apheresis yields when administered to normal platelet donors. Considerable data demonstrate the effectiveness of PEG-rHuMGDF in raising the platelet yields in apheresis donors. In the past few years, investigation into the use of thrombopoietin for ex vivo expansion of pluripotent stem cells has been extensive. Last, thrombopoietin may serve as a radioprotectant by preventing radiation-induced apoptosis of pluripotent stem cells. In the coming years, the clinical role of rhTPO, PEG rHuMGDF, and related molecules such as the thrombopoietin peptide mimetics will probably be established for both chemotherapeutic and nonchemotherapeutic indications. PMID- 10831289 TI - A structural model of the orthorhombic Ga7Mn5 derived from its HREM image AB - The base-centered orthorhombic approximant Ga7Mn5 (a = 4.547(6) nm, b = 1.256(9) nm, c = 1.482(1) nm) consisting of flattened hexagon and concave octagon (called "crown" in this paper) subunits shown by high-resolution electron microscopy (HREM) was found to coexist with the Ga-Mn decagonal quasicrystal in the Ga58Mn42 alloy. The atomic sites in the flattened hexagon subunit have been deduced recently from a structural study of the orthorhombic Ga-Mn (1/1,1/1) approximant that consists only of the hexagon subunits. Similarly, the atomic sites in the crown subunit have been deduced from another orthorhombic Ga-Mn (2/1,1/1) approximant consisting only of the crown subunits. A structural model of the orthorhombic Ga7Mn5 can thus be constructed from these structural subunits. There are 332 Ga atoms and 232 Mn atoms in a unit cell, with a composition of Mn41.2Ga58.8, satisfying the symmetry of the space group Bmm2. The simulated electron diffraction patterns, [010] HREM images, and X-ray powder diffraction pattern based on this structural model agrees mainly with the experimental observations. PMID- 10831290 TI - Recent advances in the study of quasicrystals AB - Some recent advances in the study of quasicrystals at Ames Laboratory are reviewed. In particular, growth from the melt of large, high-quality, single grain quasicrystals is described in detail for icosahedral R-Mg-Zn and decagonal Al-Ni-Co. In addition, the magnetic properties of the rare-earth-containing icosahedral R-Mg-Zn quasicrystals are discussed. PMID- 10831291 TI - Defocus convergent beam electron diffraction determination of Burgers vectors of dislocations in quasicrystals AB - Principles, method and some application examples of determining Burgers vectors of dislocations in crystals and quasicrystals by means of defocus convergent-beam electron diffraction (CBED) technique are described and reviewed and compared with contrast experiment techniques. By using defocus CBED technique, dislocation reactions during high-temperature plastic deformation of face-centered icosahedral quasicrystals have been studied. These studies lead to a preliminary understanding to the micromechanism of high-temperature plastic deformation of quasicrystals. PMID- 10831292 TI - High-resolution electron microscopy observation of a new crystalline approximant W' of Mg-Zn-Y icosahedral quasicrystal AB - A new ordered structure W' with a lattice parameter (a = 2.05 nm) about three times as large as that of the fundamental face-centered cubic W phase (a = 0.6848 nm) has been found in the Mg-Zn-Y system by means of transmission electron microscopy. The W' and W phases have the cube-to-cube orientation relationship. Moreover, the strong electron diffraction spots of the W' phase showed pseudoicosahedral symmetry, implying that it is a crystalline approximant of the Mg-Zn-Y icosahedral quasicrystal. In the high-resolution electron microscopic images of the W' phase, Penrose tiles of pentagons and boats with an edge length of a(p) = 0.481 nm can be identified. A binary tile of crown subunit has also been deduced from such a tiling. Translation domains of the W' phase have also been observed and the translation vectors at the domain boundary are: a(p), tau x a(p) and (1 + tau) x a(p), respectively, where (1 + tau) x a(p) equals to the edge length a(r) of the big obtuse rhombus of the W' phase and tau = (1 + square root of 5)/2, is the golden ratio. PMID- 10831293 TI - Twelve-fold quasicrystal and its approximant of Ta62Te38 interpreted as modulated crystals AB - A transmission electron microscopy study reveals that the twelve-fold quasicrystal and its approximant in Ta62Te38 are crystals subjected to the structure modulation. It is composed of two modulated layers rotated by 30 degrees (or 90 degrees) to each other about their normal. Structures of the twelve-fold quasicrystal and its approximant can be related by modulation waves with the same directions but with slightly different wavelengths. The modulation is considered to be due to the rearrangement of atomic vacancies as a response to the occurrence of charge density waves. PMID- 10831294 TI - An electronmicroscopic study of hexagonal phases related to quasicrystals in Zn Mg-RE alloys AB - A number of hexagonal phases with inflated/deflated a lattice parameters but equal c parameters are found to exist with compositions close to that of the icosahedral phase. Surprisingly, no cubic or orthorhombic approximants are found. The closeness of the hexagonal phases to the icosahedral phase is shown through the electron diffraction features. Through stereographic analysis of the hexagonal phases it is shown that they consist of three icosahedral units rotated by 120 degrees about a common twofold axis, forming the sixfold axis. It is suggested that this phase falls into a class of crystals closely related to both the icosahedral and decagonal quasicrystals, but are not approximants to them. These phases can instead be considered to be approximant to the hexagonal quasicrystal. PMID- 10831295 TI - Al-Cu approximants and associated B2 chemical-twinning modes AB - The Al3Cu4 alloy, with an e/a ratio of 1.86 being close to ternary Al-Cu-TM (transition metal) quasicrystals, has been chosen for the search of Al-Cu approximants. Phase structures and compositions were studied using TEM, X-ray diffraction and EPMA techniques. Two new phases were found: face-centered orthorhombic oF-Al43.2Cu56.8 (a = 0.816(6), b = 1.414(9), c = 0.999(5) nm) and body-centered orthorhombic oI-Al41.3Cu58.7 (oI, a = 0.408(3), b = 0.707(4), c = 0.999(5) nm). Their e/a ratios are the same as that of the Al-Cu-Fe icosahedral quasicrystal. Both are B2 superstructures and their unit cell components can be expressed approximately as oF-Al36Cu48vacancies12 and oI-Al8Cu12vacancies4. They both exist in twinning variants of the types 120 degrees/[001] and 180 degrees/[310]. Such twinning modes indicate that these orthorhombic phases are the decomposition products of a high-temperature parent phase epsilon2-Al2Cu3, the atomic structure of which shows pentagonal atomic arrangements. Further analysis on the twinning modes of oF and oI leads to the recognition of the chemical-twinning mode of the basic B2 structure as 180 degrees/(111)B2. This kind of chemical twinning mode is responsible for the pentagonal atomic configuration in the Al-Cu approximants as well as for the pseudo-5-fold B2 twinning. PMID- 10831296 TI - Investigation of atomic structures of diamond-like amorphous carbon by electron energy loss spectroscopy AB - Research into amorphous carbon films has been developed to such an extent that the film property can be fine tuned to mimic that of the crystalline counterparts, be it diamond, graphite, or even fullerene-like. This flexibility makes such films ideal for a wide range of applications from anti-abrasive window coating to lubricating layers on the surface of magnetic hard-disk. Not only are their mechanical properties interesting, electrically the diamond-like amorphous carbon films are also easier to dope than crystalline diamond, making them potentially a better alternative to amorphous silicon for photovoltaic devices. We will show that electron energy loss spectroscopy, in particular the carbon 1s core absorption spectroscopy, has been instrumental in revealing the nature of the bonding between carbon atoms. Such information allows microstructure models to be developed for proper understanding of the observed properties and providing scientific basis for future improvement. PMID- 10831297 TI - Experimental measurement of electron diffuse scattering in magnetite using energy filter and imaging plates AB - We have measured the diffuse scattering in magnetite as a function of temperature using the LEO 912Ohms energy-filtering electron microscope and the imaging plates. This study takes the advantage of the Koehler illumination system, energy filtering and the imaging plates for recording electron diffraction pattern over a large dynamic range. The experiment clearly shows a quantitative change in diffuse scattering distribution, which has the characteristics of one-dimensional ordering. This study clearly demonstrates the possibility for the quantitative study of diffuse scattering using electron diffraction. PMID- 10831298 TI - HRTEM image contrast of short range order in Ni4Mo AB - The high resolution transmission electron microscope (HRTEM) imaging of short range order (SRO) in Ni4Mo was investigated by means of multi-slice image simulations. The HRTEM images of Ni4Mo exhibit locally bright dot patterns corresponding to the [001] projections of the N2M2-type (chalcopyrite-like) structure. The multi-slice simulations revealed that the N2M2 patterns are rationalized as the projection patterns of the SRO structure which consists of subunit cell clusters of D1a, D022 and Pt2Mo structures. The N2M2-type image contrast appears when both the fundamental fcc lattice reflections and the 1 1/2 0 diffuse scattering of SRO contribute enough to imaging. This suggests that a good coincidence in intensity distribution between the Fourier power spectra of HRTEM images and the electron diffraction patterns is one of the conditions for the image contrast of SRO to be interpreted in terms of the projection contrast. PMID- 10831299 TI - Ordering process of Cu3Pt from short-range order state studied by in-situ TEM observation AB - The state of short-range order (SRO) and the ordering process in Cu3Pt have been studied by the in-situ transmission electron microscope observation method. In order to detect weak diffraction-patterns and weak image-contrasts, an imaging plate system and/or digital-process of video-recorded signals were applied. It is confirmed that above the critical temperature for ordering, Tc, the alloy shows diffuse scattering in its electron diffraction pattern. The appearance of the diffuse scattering indicates that the "disordered" Cu3Pt (designated as A1) has an SRO-state in thermoequilibrium. Dark field images taken by using the diffuse scattering shows a fine dot-contrast, which indicates the presence of "microdomains". The dot-contrast is flickering at higher temperatures, but the rate of flickering decreases with falling temperature and finally stops before reaching Tc. On further decreasing temperature, some of the microdomains grow up to large domains of the long period superstructure (L12-s) phase. This indicates that the transformation to L12-s via SRO proceeds without a process of nucleation even though the transformation is of a first-order. PMID- 10831300 TI - Relation between In ion ordering and crystal structure variation in homologous compounds InMO3(ZnO)m (M = Al and In; m = integer) AB - The relation between the ordering of In ions and the structure variation of homologous compounds InInO3(ZnO)13 and InAlO3(ZnO)m (m = 4, 5, and 13) have been studied by high-resolution transmission electron microscopy. It is revealed that InMO3(ZnO)m is a layered structure, consisting of InO2(1-) (In-O) and MZn(m)Om+1(1+) (M/Zn-O) layers stacked alternatively. Structure variations from the basic one, caused by the ordering of In ions in the M/Zn-O layers, are observed both in In2O3(ZnO)m and InAlO3(ZnO)m. In In2O3(ZnO)m, a modulated structure appearing as zig-zag shaped contrast in the high-resolution image was found and is considered to be caused by the ordering of In ions along the zig-zag contrast area. In InAlO3(ZnO)m, no modulated structure was found. Instead, planar defect structures appearing in Al/Zn-O layers were observed. It is shown that this defect structure is caused by the excess introduction of In ions into the Al/Zn-O layers and the ordering of these In ions. By comparing the results of InInO3(ZnO)m and InAlO3(ZnO)m, it is shown that the reasons for the In ion ordering is the discrepancy between the larger In ion size and the smaller oxygen void for M/Zn ions in M/Zn-O layers. PMID- 10831301 TI - Incommensurate valence modulation in high-Tc cuprates AB - An incommensurate modulation has been observed in a Cu-rich La2CuO4.003 crystal. It is shown that the modulation results from a periodically distributed holes lying on the CuO2 planes, and that the hole modulation may be regarded to be a kind of valence modulation. It is shown that appreciable valence modulation contrast may be generated by the mechanisms of hole scattering alone when the period of the modulation is of the order of 2 nm. PMID- 10831302 TI - Domain configurations of the "incommensurate" structures of 2H-TaSe2 and Bi2Sr2(Ca1-xLn(x))Cu2O8+delta studied by HRTEM AB - High-resolution transmission electron microscopy (HRTEM) images of the "incommensurate" structures of 2H-TaSe2 and Bi2Sr2Ca1-xLnx Cu2O8+delta (Ln: rare earth metal) are shown. They were taken from a wide specimen area with homogeneous thickness. For the former, a configuration of two domains was found by a scrutiny of HRTEM images. For the latter, many configurations of two domains were extracted from the photometric density distribution in the one-dimensional contrast modulations in HRTEM images. One domain of the two in both configurations is created by a phase slip occurring in the primary atomic displacement longitudinal wave. PMID- 10831303 TI - EELS analysis of cation valence states and oxygen vacancies in magnetic oxides AB - Transition metal oxides are a class of materials that are vitally important for developing new materials with functionality and smartness. The unique properties of these materials are related to the presence of elements with mixed valences of transition elements. Electron energy-loss spectroscopy (EELS) in the transmission electron microscope is a powerful technique for measuring the valences of some transition metal elements of practical importance. This paper reports our current progress in applying EELS for quantitative determination of Mn and Co valences in magnetic oxides, including valence state transition, quantification of oxygen vacancies, refinement of crystal structures, and identification of the structure of nanoparticles. PMID- 10831304 TI - Microstructure and composition analysis of nanostructured materials using HREM and FEG-TEM AB - The microstructure in nanostructured (NS) materials synthesized by different methods have been characterized by electron microscopy methods. NS-Pd was prepared by inert-gas condensation and in situ compacting method (IGCC), NS alloys by amorphous crystallization method (ACM) and NS-Cu and Cu100-xFe(x) alloy by mechanical alloying (MA) methods. The experimental results have revealed that different preparation techniques lead to different microstructures. The grain boundaries have ordered and disordered structures and high density of defects were frequently detected in NS-materials synthesized by IGCC and MA. For the NS alloys produced by ACM, however, the structures of GBs are similar to those in coarse-grained materials and the grains have nearly perfect crystal structure. For immiscible systems, a supersaturated Fe-Cu solid solution can be obtained by MA, but it is difficult using IGCC. PMID- 10831305 TI - Electron microscopy study of tubular crystals (BiS)1+delta(NbS2)n AB - Big crystals (mm-sized) with anomalous tubular morphology were synthesised in the Bi-Nb-S system. Their structural study by electron microscopy and related techniques revealed that they can be described as misfit layer structures, a type of composite modulated structures. The misfit monolayer approximately BiNbS3 and bilayer approximately BiNb2S5 phases appear as well in the preparation but with lamellar morphology as it is typical for these kind of compounds. They consist of periodical intergrowth of pseudotetragonal layers BiS (Q) with pseudohexagonal layers NbS2 (H). approximately BiNbS3 has a stacking sequence ...Q,H,Q,H,... and approximately BiNb2S5 has a stacking sequence ...Q,H,H,Q,H,H,... Backscattered electron imaging, wavelength dispersive X-ray spectroscopy and transmission electron microscopy of transversal cross-sections of single tubular crystals showed that these crystals present a strong compositional and inter-laminar stacking disorder along the tube radius. This disorder is suggested to be the cause for the original wrapping of the layers that gives rise to the tubes. Besides the disordered areas, some ordered slabs have been found with stacking sequences corresponding to binary 3R-NbS2 (approximately 6 A), approximately BiNb2S5 (approximately 17.4 A) (which dominate in the crystal) and to a new related phase approximately BiNb4S9 with a stacking sequence ...Q,H,H,H,H,Q,H,H,H,H,... and a periodicity of approximately 29.2 A. PMID- 10831306 TI - A TEM and XRD study of the SbNbSe3 misfit layer structures AB - Two new misfit layer structures have been synthesized within the Sb-Nb-Se system. Powder X-ray diffraction and electron microscopy techniques (electron diffraction, HREM, XEDS) have been used to determine the nature of their structure. According to TEM and XEDS data (for more than 15 crystals studied) both phases are monolayer type, i.e. (SbSe)1+delta (NbSe2). Electron microscopy reveals a composite modulated structure that consists of the periodical intergrowth of a pseudotetragonal SbSe layer, denominated as Q, and a pseudohexagonal layer NbSe2, denominated as H. Both layers fit along b, stack along c and do not fit along a (misfit) giving rise to an incommensurate modulation along this direction. The two phases differ in the symmetry of the Q layers being in one case orthorhombic (for delta = 0.17) and monoclinic in the other (for delta = 0.19). After the characterization of the sample by electron microscopy the unit cells of the basic layers could be refined for both phases by powder X-ray diffraction: aQ = 5.824(2) A, bQ = 5.962(5) A, cQ = 23.927(6) A, alpha = 90 degrees, beta = 90 degrees and gamma = 90 degrees and aH = 3.415(5) A, bH = 5.962(6) A,, cH = 11.962(1) A, alpha = 90 degrees, beta = 90 degrees and gamma = 90 degrees for the orthorhombic phase; aQ = 5.844(2) A, bQ = 5.981(1) A, cQ = 23.919(5) A, alpha = 90 degrees, beta = 90 degrees and gamma = 96.00(3)degrees and aH = 3.439(1) A, bH = 5.994(2) A, cH = 11.956(3) A, alpha = 90 degrees, beta = 90 degrees and gamma = 90 degrees for the monoclinic phase. The phase with the monoclinic Q-sublattice often appears as twinned crystals. The more abundant crystals are disordered intergrowths of both monolayer phases. PMID- 10831307 TI - HRTEM investigation of mesoporous molecular sieves AB - High resolution transmission electron microscopic (HRTEM) studies of some typical mesoporous molecular sieves, such as MCM-41, SBA-2 and STAC-1, etc. are presented. Since the materials consist of amorphous silica and their unit cell dimensions are in a range of 1.5 to over 10 nm, the conventional X-ray diffraction method gives us very limited information about the detailed structures. On the other hand, HRTEM has been found to be the most powerful technique to detect the local structures and to image metal clusters inside the channels of these materials. PMID- 10831308 TI - General traits of personality and affectivity as predictors of satisfaction in intimate relationships: evidence from self- and partner-ratings. AB - Self- and partner-ratings on trait affect and the Big Five were obtained from 74 married and 136 dating couples. The relationship satisfaction of each person (the "target") was correlated with four sets of ratings: (a) target's self-rated personality, (b) target's partner-rated personality, (c) partner's self-rated personality, and (d) partner's target-rated personality. Self- and partner ratings of the target's personality yielded very similar results. Negative and positive affectivity were consistent predictors of satisfaction in both samples. Conscientiousness and agreeableness were reliably related to satisfaction in the dating couples, whereas extraversion consistently correlated with satisfaction in the married couples. These traits jointly predicted as much as 34% (self-ratings) and 26% (partner-ratings) of the variance in satisfaction. In contrast, the partner's personality played a lesser role in satisfaction. PMID- 10831309 TI - The Riverside Behavioral Q-sort: a tool for the description of social behavior. AB - The Riverside Behavioral Q-sort (RBQ) is a flexible technique for gathering a wide-ranging description of the behavior of individuals in dyadic social interaction. Ratings of RBQ items can attain adequate reliability to reflect behavioral effects of experimental manipulations and to manifest meaningful correlations with a variety of personality characteristics. The RBQ's flexibility, validity, and relative ease of use may facilitate the more frequent inclusion of behavioral data in personality and social psychology. PMID- 10831310 TI - Subjective overachievement: individual differences in self-doubt and concern with performance. AB - We discuss the construct of doubt about one's competence and suggest that doubt can have myriad consequences (e.g., self-handicapping, defensive pessimism). We focus on the effect of self-doubt when it is combined with a concern with performance and assert that this combination leads to the phenomenon of subjective overachievement. In two studies, we present a new 17-item Subjective Overachievement Scale (SOS), which includes two independent subscales measuring individual differences in self-doubt and concern with performance. The first study, consisting of two large samples (Ns = 2,311 and 1,703), provides evidence that the scale has high internal consistency and a clear two-factor structure. Additionally, the subscales have adequate test-retest reliability (Ns = 67 and 115). A second study reveals that the SOS has good convergent and discriminant validity. Both subscales are unrelated to social desirability but exhibit the predicted patterns of associations with other related constructs. The Concern with Performance Subscale is correlated with achievement motivation, whereas the Self-Doubt Subscale is correlated with scales assessing negative affectivity (e.g., self-esteem, social anxiety) and other self-related strategies associated with concerns about one's competence (e.g., self-handicapping, defensive pessimism, impostor phenomenon). The SOS, which combines the two subscales, appears to tap a unique strategy that individuals may use to deal with doubts about their own competence. PMID- 10831311 TI - Love and anger in romantic relationships: a discrete systems model. AB - In a study of 124 dating couples, we tested a discrete systems model of the functions of two emotion systems in romantic relationships: love and anger/upset. This model posits that the operation of these systems reflects adaptations shaped by natural selection to solve different adaptive problems. Accordingly, we hypothesized that the love and anger/upset emotion systems would be largely independent in the classes of information they track in romantic relationships, in the psychological mechanisms that process that information, and in the resultant behavior generated. Consistent with the discrete systems model, and in contrast to a competing "crossover" model, differences across relationships in feelings of love covaried with differences in strategic facilitation but not in strategic interference by partners. Similarly, differences in feelings of anger/upset during conflict covaried with differences in strategic interference but not strategic facilitation. In turn, feelings of love predicted commitment promoting behavior but not partner-directed aggression, whereas levels of anger/upset predicted aggression but not commitment. As also predicted by our model, the love and anger/upset emotion systems converged to predict relationship satisfaction. PMID- 10831312 TI - The causes and costs of crime and a controversial cure. AB - In spite of recent modest decreases, the epidemic of crime that began in the United States in the early 1960s has left us with a rate of violent crime that is still some 300% higher than it was 40 years ago. The usual suspects--poverty, the easy availability of street drugs and handguns, violent television programs- cannot account for this ominous trend. The fact that African Americans are responsible for nearly half of this violence, although they constitute only about one-eighth of the U.S. population, is the principal reason why the great improvement in race relations made over the past half-century has reached an asymptote. White fears, and Black resentment of these fears, are a grave threat to further progress. It is argued here that the main reason for this epidemic of crime and violence is the rapid recent increase in the proportion of the young people aged 15 to 24 who have grown up unsocialized. It is argued further that most of these feral youngsters are sociopaths, defined as genetically normal children whose failure of socialization was due to their being domiciled with an immature, overburdened, unsocialized, or otherwise incompetent parent or parents. PMID- 10831313 TI - Crime, criminals, and cures: medical model revisited. AB - David Lykken's target article assesses the causes of crime and advocates a controversial "cure"--parental licensure. Although Lykken gets many of the facts about criminals right, ultimately the disease metaphor breaks down. Crime requires three things--motivated offenders ("criminals"), suitable targets or victims, and the absence of capable guardians to prevent the act. Typical of medical model approaches, failure to consider the convergence in time and space of the three necessary elements for crime results in a misdiagnosis. In this invited commentary, I briefly note three reasons why Lykken's cure, along with the medical model in general, is unlikely to bear fruit. PMID- 10831314 TI - Toward voluntary parenthood. AB - David Lykken's proposal to license married parents for child rearing, and to deny the same opportunity to single and inept parents, springs from his deep concern for millions of youngsters cruelly subjected to abusive and neglectful rearing circumstances. Children from such inadequate homes grow up to have high rates of school failure, criminality, and drug addiction. The problem is clear, but Lykken's remedies of mandated marriage and parental licensure are unacceptable in U.S. society, where our reproductive rights are fortunately protected by our Constitution. As a devoted proponent of reproductive rights, I propose a legally and morally acceptable proposal to the same end. Increasing women's effective control of reproduction and moving toward entirely voluntary parenthood will accomplish the same goals without compromising our civil liberties. PMID- 10831315 TI - The outcome of parenting: what do we really know? AB - Lykken's proposal that people should be required to apply for a license before they can become parents is based on his belief that the increase in crime over the past 40 years is due largely to the increase in the number of children reared in fatherless homes. I argue that the absence of a father is a correlate of criminal behavior in the offspring but not its cause. Other correlations that are usually interpreted as the effects of good or bad child-rearing practices are similarly ambiguous and subject to other explanations. I conclude that the increase in crime over the past 40 years cannot be blamed on the absence of fathers or inept child rearing, and offer two alternative hypotheses. PMID- 10831316 TI - Regulation of TRAIL-induced apoptosis by transcription factors. AB - The tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) is a newly identified member of the TNF family. Unlike many other members of the TNF family, TRAIL selectively induces apoptosis of tumor cells, but not normal cells. The mechanisms whereby TRAIL-induced apoptosis is regulated in various cell types are not clear. We report here that the peroxisome proliferator-activated receptor (PPAR)-gamma and nuclear factor (NF)-kappaB play distinct roles in regulating TRAIL-induced apoptosis. Activation of PPAR-gamma by its agonist pioglitazone significantly enhanced TRAIL-induced apoptosis. This was associated with inhibition of proliferation and cell cycle progression. On the other hand, inhibition of NF-kappaB by sulfasalazine also significantly enhanced TRAIL induced apoptosis. These results strongly suggest that while transcription factor PPAR-gamma promotes TRAIL-induced apoptosis, NF-kappaB inhibits it. Thus, PPAR gamma agonists and NF-kappaB inhibitors are potent enhancers of TRAIL-induced apoptosis. PMID- 10831317 TI - The epithelial mucin, MUC1, is expressed on resting T lymphocytes and can function as a negative regulator of T cell activation. AB - MUC1 is a mucinous glycoprotein which is normally expressed on the surface of a variety of epithelia and aberrantly overexpressed on some human tumors. In this report, we demonstrate that the epithelial mucin, MUC1, is expressed on resting human peripheral blood T cells and two leukemia T cell lines, Jurkat and Hut 78. Crosslinking of MUC1 on peripheral blood T cells by plate-bound anti-MUC1 (DF3-P) antibody inhibits cell proliferation, IL-2 and GM-CSF production, and up regulation of the IL-2 receptor upon anti-CD3 stimulation. Induction of IL-2 production by Jurkat and HUT 78 is also suppressed and cannot be reversed by the addition of anti-CD28 mAb. These findings suggest that MUC1 can be a potent negative regulator for T cell activation at the resting stage. PMID- 10831318 TI - Neutrophil chemotaxis and superoxide production are induced by cross-linking FcgammaRII receptors. AB - Neutrophils express two types of receptor for the Fc region of IgG, FcgammaRII and FcgammaRIIIB. Via these receptors, neutrophils bind IgG complexes that contain more than one IgG molecule. This binding activates functional processes, such as the respiratory burst and chemotaxis. Neutrophils were treated with biotinylated anti-Fc receptor monoclonal antibodies and chemotaxis toward streptavidin, a cross-linking agent, was determined. Cross-linking FcgammaRII and not FcgammaRIIIB induced neutrophil chemotaxis. Superoxide production in response to immobilized anti-Fc receptor antibodies was also examined. Anti-FcgammaRII Fab bound to ELISA plates induced superoxide production, while anti-FcgammaRIIIB Fab did not. Pretreatment of neutrophils with anti-FcgammaRII Fab reduced superoxide generated by immobilized anti-FcgammaRII antibody. The data demonstrate that FcgammaRII and not FcgammaRIIIB are responsible for neutrophil chemotaxis and superoxide production upon Fc receptor activation. PMID- 10831319 TI - Antigen-dependent cytokine mRNA expression by individual rhesus macaque T helper cells by flow cytometry. AB - Specific patterns of cytokine secretion by CD4(+) T helper (Th) cells determine the nature of immune effector responses. Using a multiparameter, flow cytometric fluorescent in situ hybridization (FISH) assay that detected cytoplasmic mRNA within intact cells, we assessed antigen-specific cytokine expression in rhesus macaque Th cells. In the peripheral lymphocytes of immunized rhesus macaques, FISH detected antigen-induced cytokine gene expression in single Th cells. Analysis of simultaneous cytokine expression by single cells demonstrated that the recall immune response consisted of Th cells expressing either a Th1 (IL 2(+)/IFN-gamma(+)) or a Th2 (IL-4(+)/IL-6(+)) cytokine pattern. In addition to the classic Th subsets, Th cells expressing only one of two Th1 or Th2 defining cytokines were common following antigen restimulation. The data gathered with the FISH assay suggest that, in primates, the immune response to recall antigens consists of nonclassic Th cells, as well as a mixture of polarized Th1 and Th2 T cells. PMID- 10831320 TI - Differential regulation of CD40-mediated human B cell responses by antibodies directed against different CD40 epitopes. AB - Ligation of CD40 using anti-CD40 or soluble CD40-ligand activates numerous intracellular kinases which transduce signals to the nucleus. The nature whereby these signaling events are coupled to distal functional events in B cells is poorly understood. In this study, using anti-CD40 monoclonal antibodies which recognize different epitopes on CD40, we compare the ability to activate the stress-activated protein kinases (SAPK) such as c-Jun NH(2) terminal kinase and p38 in human B cells with CD40 function. Activation of the SAPK pathway correlated with levels of activation of Rel/NF-kappaB transcription factors, but did not appear to be associated with rescue from anti-IgM induced apoptosis by suppressing caspase (CPP32) activity. Somewhat surprisingly, in the presence of IL-4, those antibodies to CD40 which failed to activate SAPK were most active in IgE production. IgE production was augmented in the presence of wortmannin. These studies suggest that rescue from apoptosis and IgE production mediated via CD40 may be independent of SAPK activation, induction of Rel/NF-kappaB, or suppression of CPP32 and that IgE production is, at least in part, regulated by signaling pathways that are dependent on phosphatidylinositol 3-kinase. PMID- 10831321 TI - Lipoproteins from Borrelia burgdorferi applied in liposomes and presented by dendritic cells induce CD8(+) T-lymphocytes in vitro. AB - Borrelia burgdorferi (Bb) is the tick-borne etiologic agent of Lyme borreliosis, which has many aspects of autoimmune diseases. Bb is unable to recycle synthesized membrane lipids and lipoproteins. Consequently, a large amount of liposome-like vesicle (Bb-blebs) is shed from the outer bacterial membrane. The influence of Bb-blebs on the cellular immune response is not yet known. As a Bb blebs model, we established standardized Bb-liposomes, produced from freshly extracted lipids and lipoproteins of live Bb. Bb-liposomes were incorporated via nonendocytotic mechanisms by different human cell types, namely dendritic cells (DC), lymphocytes, and fibroblasts, as visualized by immunofluorescence and transmission electron microscopy. Bb-liposomes were localized in the cytosol and in the nucleus of the cells. With this in mind, we generated in vitro Bb-specific T-cells from nonadherant peripheral blood mononuclear cells by use of Bb liposomes loaded autologous DC. More than 95% of those T-cells were CD8(+) and they killed autologous Bb-liposome-loaded T-cell blasts. These results suggest that Bb-blebs may be responsible for the autoimmune-like appearance of Lyme disease. PMID- 10831322 TI - B7.1 on human carcinomas: costimulation of T cells and enhanced tumor-induced T cell death. AB - Human squamous cell carcinomas of the head and neck (SCCHN) do not express the costimulatory molecules B7.1 or B7.2 in situ or in culture. Transduction of B7.1( ) SCCHN cells with the retroviral B7. 1 and neo(r) genes resulted in the expression of high levels of the transgene in these tumor cells. When B7.1(+) SCCHN cells were used as stimulators of autologous or allogeneic PBL in mixed lymphocyte-tumor cultures (MLTC), T-cell proliferation and generation of antitumor effector T cells as well as levels of their lytic activity were significantly increased. At the same time, a proportion of activated T cells seen to undergo apoptosis was found to be significantly higher upon coincubation with B7.1(+) SCCHN than with B7.1(-) SCCHN. Both B7.1(+) and B7.1(-) SCCHN cells were found to express FasL on the cell surface and in the cytoplasm, as well as mRNA for FasL and mRNA for TRAIL. However, expression of the B7.1 transgene did not lead to increased expression of FasL protein on tumor cells. Yet, up to 50% of activated CD28(+) allogeneic T cells, which were CD95(+), showed evidence of DNA fragmentation in JAM and TUNEL assays upon incubation with an excess of B7.1(+) SCCHN for 24 h. Tumor-induced T-cell death was equally and only in part blocked by anti-Fas antibodies in both B7.1(+) and B7.1(-) MLTC. While surface expression of B7.1 molecules on SCCHN cells enhanced T-cell costimulation via B7.1-CD28 interactions, it did not rescue activated T cells from tumor-induced apoptosis. The outcome of MLTC under these conditions was dependent on the ratio of tumor to T cells. Thus, in the presence of an excess of B7.1(+) tumor cells, activated T cells showed increased sensitivity to apoptosis which did not appear to be Fas/FasL mediated. These data are important for the development of B7.1 gene therapy and efforts directed at the generation of effector cells in MLTC. PMID- 10831323 TI - A native soluble form of CTLA-4. AB - CTLA-4 is an immunoregulatory receptor expressed on the surface of activated T and B lymphocytes. The counterreceptors for CTLA-4 are the B7 family molecules. We describe alternatively spliced mRNAs expressed in hematolymphoid tissues of humans, mice, and rats that lack the transmembrane domain coded by exon 3 of the CTLA-4 gene. These alternate transcripts were detected by RT-PCR in B cells and resting T cells of both the CD4 and the CD8 phenotype. Activation of human blood mononuclear cells with PHA or anti-CD3 + anti-CD28 monoclonal antibodies appears to effect a decrease in the amount of the alternative transcript relative to the full-length transcript. Recombinant sCTLA-4 is a B7-binding protein and has immunomodulatory effects as measured by inhibition of the mixed leukocyte response. Human serum contains immunoreactive material consistent with a native soluble form of CTLA-4. PMID- 10831324 TI - Southeast Asia, promotability and dioxin's relationship to cancer incidence in operation ranch hand veterans. AB - Significant increases in systemic, melanoma, and presumedly total skin cancers are observed in Operation Ranch Hand veterans unexposed to dioxin. The spiked increase of excess systemic cancers and skin cancer incidence is the pattern expected from a 2- to 3-year exposure to cancer initiators in Southeast Asia during tours of duty coupled with brief or full promotability of initiated cells later in the United States. A discussion of promotability is undertaken. The exposure of these airmen to both Southeast Asia and dioxin is associated with a significant reduction of elevated systemic cancers and total skin cancers. PMID- 10831326 TI - The snail as a target organism for the evaluation of industrial waste dump contamination and the efficiency of its remediation. AB - The diagnosis of contamination and the efficiency of remediation of an industrial waste dump (IWD) were done before and after remediation. For this study, two species of snails were used for passive and active biomonitoring: a nonnative species, Helix pomatia and young garden snails (Helix aspersa aspersa) of standardized rearing (age, 2 months). Bioaccumulation analysis of pollutants (cadmium, nickel, iron, lead, chromium, manganese, copper, and zinc) in viscera of the snails gave data on the evolution of the biotop biodisponibility over time. Those data allowed the conclusion that the snails were useful for passive and active biomonitoring. Comparison of two contamination grids corroborated previous conclusions. Furthermore, the results obtained indicated the efficiency of the remediation technique used for the IWD. PMID- 10831325 TI - Variations in metallothionein, Zn, Cu, and Fe concentrations and ceruloplasmin activity in pregnant rat dams and their fetuses. AB - Growing rat fetuses have great need for Zn, Cu, and Fe. In fetal livers (FL) large accumulations of Zn and Cu connected with increased metallothionein (MT) synthesis take place. In dams, serum changes in metals concentrations with increased ceruloplasmin (Cp) activity are observed. The aim of this study was to determine (1) mutual relationships in the accumulation of MT, Zn, Cu, and Fe in fetal livers; (2) changes in Zn, Cu, and Fe concentrations in dam serum; and (3) the day with the maximum Cp activity. Sections of rat dams were taken on 16th 21st day of gestation, twice a day, and MT, Zn, Cu, and Fe concentrations in liver, spleen, kidneys, and placenta of dams and in liver and brain in fetuses were determined. In fetal livers high correlations between MT and Zn and between Zn and Cu were obtained. The investigated Cp activity was always high, reaching its maximum on the 20th day and minimum on the 21st day. Significant correlation between Cp activity and Cu concentration in dam serum was also revealed. In conclusion it is suggested that Zn accumulation in FL is strictly connected with MT synthesis but Cu content in FL is rather dependent on Cp activity in dam serum. Iron accumulation in fetal livers is connected with the diminution of iron concentrations in dam serum. PMID- 10831327 TI - Derivation of predicted No-effect concentrations for lindane, 3, 4 dichloroaniline, atrazine, and copper. AB - Environmental risk assessment is a key feature of regulations controlling the placing of new, and the maintenance of existing, chemicals products in the market place. For example, European Commission Directive 93/67/EC on Risk Assessment for New Notified Substances and Commission Regulation (EC) No. 1488/94 on Risk Assessment for Existing Substances requires that risk assessments be carried out for new and existing substances in the European Community. The process of environmental risk assessment seeks to determine the balance of probability of species and communities being damaged by chemical releases. The process relies upon a valid estimation of a predicted environmental concentration (PEC) in relevant environmental compartments and a predicted no effect concentration (PNEC) below which the organisms present in that compartment are unlikely to be significantly affected. If the PEC exceeds the PNEC there is a potential for damaging effects to occur. This article focuses on the determination of PNECs for risk assessment. Methods for determining a PNEC described in OECD Monograph 26 (1989, Report of the OECD Workshop on Ecological Effect Assessment, Paris, France, have been applied to data derived for the four chemicals lindane, 3,4 dichloroaniline, atrazine, and copper in a series of collaborative research projects funded by the European Commission. PMID- 10831328 TI - Dissipation of lindane and fenvalerate residues in chickpea (Cicer arietinum L.) under Indian climatic conditions. AB - Dissipation behavior of lindane 20 EC (400 and 800 g a.i./ha) and fenvalerate 0.4% dust (75 and 150 g a.i./ha) applied on chickpea (var. H 208) for controlling pod borer under field conditions during rabi 1994-1995 at pod initiation stage was studied. Grains from lower dose of lindane at harvest contained residues below maximum residue limit value (0.25 microg g(-1)), whereas no measurable amount of fenvalerate residues were found in grains of both doses at harvest. A waiting period of 7 days for lindane and 1 day for fenvalerate is suggested from point of hazards due to residues, when applied at recommended dose. The residues calculated from consumption of chickpea (60 g/person/day) is much below the maximum permissible intake of lindane and fenvalerate; hence the recommended doses can be considered safe from point of view of hazards to humans. PMID- 10831329 TI - DNA adducts in perch (Perca fluviatilis) living in coastal water polluted with bleached pulp mill effluents. AB - DNA adducts were analyzed by (32)P-postlabeling in liver and intestine of perch (Perca fluviatilis). Fish were collected in the receiving water of a bleached kraft pulp mill at Norrsundet, Sweden, on the coast of the Bothnian Sea, and in three different reference areas. Sampling was carried out the last week in September 1993, 1995, and 1997. Since 1984/1985 the pulping process at Norrsundet has been successively modified and an external waste-water treatment has been installed, resulting in lowered and altered effluent discharges. Nevertheless, perch captured in the water area closest to the mill (2 km) had significantly elevated levels of aromatic/hydrophobic DNA adducts in both liver and intestine when compared to perch captured 8 km away and in the reference areas. Autoradiograms indicated a diagonal zone consisting of multiple overlapping adducts, a pattern typical of exposure to a complex mixture of aromatic/hydrophobic genotoxic substances. No significant difference in adduct levels was detected between the years. Results from this study reveal that fish in the area closest to the mill are exposed to substances with a genotoxic potential. PMID- 10831330 TI - Fish uptake of inorganic and mucus complexes of lead. AB - Carassius auratus were exposed to lead in synthetic water. Mucus secreted from the fish was measured and characterized for its lead binding capacity. The increase in mucus concentration in the water depended on secretion and degradation rates and the former was speeded up by lead. The complexation between mucus and lead can be described based on a single-site interaction model. Lead speciation in the system with or without mucus was calculated using chemical equilibrium modeling. The dominant species of lead was PbCO(3)(0). Other major species included Pb(OH)(+), Pb(OH)(2)(0), Pb(CO(3))(2)(2-), Pb-mucus, and Pb(2+), among which free lead and lead hydroxo complexes were available to fish uptake. PMID- 10831331 TI - Fiber optic biosensor using Chlorella vulgaris for determination of toxic compounds. AB - A new biosensor is constructed for the detection of some herbicides based on kinetic measurements of chlorophyll-a fluorescence in Chlorella vulgaris cells. The microalgae are immobilized on removable membranes placed in front of the tip of an optical fiber bundle inside a homemade microcell. C. vulgaris was easily cultivated in laboratory and very sensitive to herbicides that effect the photosynthesis process. The response of the algal biosensor is studied in terms of detection limits, reversibility, and long-term activity. The effects of temperature and pH are also reported. The biosensor can be used to measure the concentration of a toxic chemical in the form of a single drop or dissolved in a continuous flow. The detection of 0.1 microg small middle dotL(-1) of a single herbicide as is required by European Community legislation for drinking water is possible with this algal biosensor especially for atrazine, simazine, and diuron. PMID- 10831332 TI - Fate and the biochemical effects of 2,4,6-trinitrotoluene exposure to tiger salamanders (Ambystoma tigrinum). AB - Biotransformation, metabolic enzyme profiles, and the glutathione antioxidant system in tiger salamanders (Ambystoma tigrinum) from a 14-day exposure to 2,4,6 trinitrotoluene (TNT) in situ were examined. Concentrations of parent compound and metabolites were measured in skin, kidney, and liver tissue. In addition, cytochrome P450 (P450) and cytochrome b5 content and their dependent isozyme activities, ethoxyresorufin O-dealkylation, pentoxyresorufin O-dealkylation, and the glutathione antioxidant system in the skin, liver, lung, kidneys, and blood were evaluated. Considerable differences were found in relative concentrations of TNT and its metabolites in the skin, relative to the liver and kidney. Trace amounts of TNT were detected only in the skin and liver of exposed animals while one of the secondary reduction metabolites, 2,6-diaminonitrotoluene, was found only in liver and kidney. Differences in the metabolite concentrations between systemic organs (liver, kidneys) and the skin suggest that the skin may be important in the primary reduction of TNT. In addition, measurable levels of these basal enzyme indicators were detected; yet of those evaluated only hepatic P450 content was affected by TNT exposure. The qualitative and quantitative differences in TNT and its metabolites in tissues suggest the fate and metabolism of the TNT in salamanders. Furthermore, results indicate that tiger salamanders possess considerable levels of xenobiotic metabolizing and antioxidant enzymes in these tissues but are not sensitive indicators of TNT exposure. PMID- 10831333 TI - Effects of spray-irrigated wastewater effluent on temporary pond-breeding amphibians. AB - Comparison was made of abiotic conditions, amphibian reproductive outputs, and the survival of embryonic and larval amphibians in wastewater effluent-irrigated and natural temporary ponds in an area in Centre County, Pennsylvania that has been spray-irrigated with secondarily treated, chlorinated wastewater effluent from The Pennsylvania State University for approximately 14 years. Three species of temporary pond-breeding amphibians were studied: wood frogs (Rana sylvatica LeConte), Jefferson salamanders (Ambystoma jeffersonianum Green), and spotted salamanders (A. maculatum Gravenhorst). Comparisons of physico-chemical parameters in 10 wastewater-irrigated and 10 natural temporary ponds over 19 weeks in 1997 indicated that wastewater -irrigated ponds had significantly higher median conductance, pH, ?Na, ?K, ?Ca, ?Mg, and ?N-NO(3) and lower ?dissolved oxygen. Many of the wastewater-irrigated ponds supported large mats of duckweed (Lemna spp.) that completely blanketed the pond's surface by mid-May. There were significantly fewer egg masses of all three species in wastewater-irrigated ponds than in natural ponds in both 1997 and 1998. In situ egg hatching success and larval survival (over a 6-day period) of all species was lower in wastewater irrigated ponds than in natural ponds. Cumulatively, these studies suggest that wastewater effluent irrigation may impact amphibian populations by reducing the survival of amphibian eggs and larvae. PMID- 10831334 TI - Inhibition of ovarian growth by cadmium in the fiddler crab, Uca pugilator (Decapoda, ocypodidae). AB - The effect of cadmium chloride (1 mg/L) on oocyte growth of the fiddler crab, Uca pugilator, was studied during the slow vitellogenesis phase of ovarian maturation of this carb. In vivo experiments were done with both intact and eyestalkless crabs. The intact cadmium-exposed crabs exhibited a significantly lower oocyte diameter than the controls at the end of the 2-week exposure period, but no significant differences were detected among the eyestalkless crabs, suggesting that the effect of cadmium could be on the sinus gland in the eyestalks, increasing secretion of the gonad-inhibiting hormone (GIH). To test this hypothesis, in vitro experiments were done, incubating pieces of ovary with and without eyestalk tissue, in the presence of thoracic ganglion, needed for oocyte growing due to the secretion of the gonad-stimulating hormone (GSH). Only when eyestalk tissue was present in the incubation media was oocyte growth inhibited by cadmium compared to the controls. These results strongly suggest that cadmium acts at least in part to increase the secretion of GIH from the sinus gland. GIH could then in turn act by (a) inhibiting secretion of GSH by the thoracic ganglion, (b) directly inhibit the oocytes, or (c) both (a) and (b). PMID- 10831335 TI - Effects of 2,4-dichlorophenol on the net-spinning behavior of hydropsyche slossonae larvae (Trichoptera; Hydropsychidae), an early warning signal of chronic toxicity. AB - In order to assess the potential of hydropsychid capture net anomalies as a bioindicator of chronic toxicity in streams and rivers, the effects of 2,4 dichlorophenol (2,4-DCP) exposure on the net-spinning behavior of Hydropsyche slossonae were examined for anomalies after 0, 5, 10, 15, and 20 exposure days to gradient concentration of 2,4-dichlorophenol. The net-spinning behavior was significantly affected when larvae were exposed to 1.0, 10, 25, and 50 microg small middle dotL(-1), as expressed by the occurrence of two distinct abnormalities. The first one was a distortion of the midline meshes, where the normal diamond-shape structure is disrupted and the meshes are separated by extra strands (called "midline anomaly). The second aberration observed was called "chaotic net, where the nets are highly irregular without any real structure or well-defined areas. A good correlation was found between the chaotic net frequencies and the reduction of ATP concentrations in the larvae, indicating possible uncoupling effects of 2,4-DCP on the oxidative phosphorylation process. Toxicity curves demonstrate that the sensitivity threshold of chaotic net frequencies ranged from 3.5 to 7 microg small middle dotL(-1), which is highly sensitive compared with other sublethal effects of 2,4-DCP on other aquatic species. PMID- 10831336 TI - Iron ore mines leachate potential for oxyradical production. AB - The ecotoxicological effects of mining effluents is coming under much greater scrutiny. It appears necessary to explore possible health effects in association with iron ore mining effluents. The present results clearly demonstrate that iron ore leachate is not an inert media but has the potential to induce lipid peroxidation. Peroxidation was assessed by measuring oxygen consumption in the presence of a reducing agent such as ascorbate or NADPH and a chelator such as EDTA. Labrador iron ore is an insoluble complex crystalline material containing a mixture of metals (Fe, Al, Ti, Mn, Mg,ellipsis, ) in contrast to the iron sources used for normal lipid peroxidation studies. The metal of highest percentage is iron (59. 58%), a metal known to induce oxyradical production. Iron ore powder initiated ascorbic acid-dependent lipid peroxidation (nonenzymatic) in liposomes, lipids extracted from rat and salmon liver microsomes, and intact salmon liver microsomes. It also revealed an inhibitory effect of NADPH-dependent microsomes lipid peroxidation as well as on NADPH cytochrome c reductase activity. However, nonenzymatic peroxidation in rat liver microsomes was not significantly inhibited. Cytochrome P450 IA1- and IIB1-dependent enzymatic activities as well as P450 levels were not affected. The inhibition could be due to one of the other components of iron ore leachate (Mn, Al,ellipsis, ). These effects of iron-ore leachate indicate that a potential toxicity could be associated with its release into lakes. Further studies are necessary to explore in vivo effects on aquatic animals. PMID- 10831337 TI - Acute toxicity to rats and trout with a focus on inhalation and aquatic exposures. AB - Due to limitations in toxicity data, extrapolations are often performed within and between species. In this context, median lethal concentration (LC(50)) and dose (LD(50)) for 231 chemicals, along with physicochemical factors, were log transformed and compared across several exposure routes in rats (i.e., oral, dermal, intraperitoneal, inhalation, intravenous, subcutaneous) and rainbow trout (i.e., aquatic, oral, dermal, intraperitoneal). Molecular weight (MW), octanol water partition coefficient (K(ow)), vapor pressure (VP), and water solubility (S) were intercorrelated (Bonferroni P<0.05). MW (7 of 10 cases) and K(ow) (3 of 10 cases) were inversely correlated to toxicity endpoints, whereas VP (5 of 10 cases) was positively correlated (Bonferroni P< 0.05). Interspecies correlations were observed in toxicity endpoints over multiple exposure routes (10 of 24 cases), in addition to intraspecies correlations within rat (13 of 15 cases) and within trout (3 of 5 cases) (Bonferroni P<0.05). It was hypothesized that rat inhalation LC(50) and trout LC(50) would be associated, since air and water exposures correspond to each organism's respiratory medium. This hypothesis was supported by their correlation (r=0.68, Bonferroni P<0.05), characterized by a median molar ratio near 1, and further analyzed via regression methods. Comparison of acute toxicity endpoints can, therefore, assist in identifying appropriate extrapolations, likely based on similar toxicokinetics. PMID- 10831338 TI - Relationship between toxicokinetics of carbaryl and effect on acetylcholinesterase activity in Pomacea patula snail. AB - The 96-h LC(50) value of carbaryl was 14.6 microg/mL for the snail Pomacea patula. Organisms were exposed for 72 h to a low sublethal concentration (0.1 of LC(50)) using a semistatic contamination system; bioconcentration and elimination experiments were performed evaluating simultaneously acetylcholinesterase (ACHase) activity. The inhibition of the digestive gland ACHase reached 76% when the carbaryl concentration in tissue was 3.2 microg/g. The increased enzyme inhibition was observed concomitantly with the bioconcentration of carbaryl until 7 h. ACHase inhibition was linearly dependent on the uptake and bioconcentration of carbaryl (r(2)=0.87). The transfer of snails to carbaryl-free water after 72 h of exposure was followed by rapid monophasic elimination with a half-life of 1.0 h. However, ACHase activity levels never returned to control values. These results revealed that the bioconcentration might play a critical role in contributing to the toxicity of carbaryl. PMID- 10831339 TI - Papers to appear in environmental research section A PMID- 10831340 TI - Is transvaginal ultrasound effective for screening asymptomatic women for the detection of early-stage epithelial ovarian carcinoma? PMID- 10831341 TI - The efficacy of transvaginal sonographic screening in asymptomatic women at risk for ovarian cancer. AB - OBJECTIVE: The purpose of this study was to determine the efficacy of annual transvaginal sonography (TVS) as a screening method for ovarian cancer. METHODS: Annual TVS screening was performed on 14, 469 asymptomatic women from 1987 to 1999. Eligibility criteria included (1) all women >/= 50 years of age and (2) women >/= 25 years of age with a family history of ovarian cancer. Ovarian volume was calculated using the prolate ellipsoid (length x height x width x 0.523). An abnormal sonogram was defined by (1) an ovarian volume >10 cm(3) in postmenopausal women or >20 cm(3) in premenopausal women or (2) a papillary or complex tissue projection into a cystic ovarian tumor. All women with abnormal TVS had a repeat sonogram in 4-6 weeks. Patients with a persistently abnormal second screen had a serum CA-125 determination, tumor morphology indexing, and Doppler flow sonography, and were advised to have surgical tumor removal. RESULTS: One hundred eighty patients with persisting TVS abnormalities underwent exploratory laparoscopy or laparotomy. Seventeen ovarian cancers were detected: 11 Stage I, 3 Stage II, and 3 Stage III. Only three patients with Stage I cancers had a palpable ovarian mass on clinical examination. All patients with Stage I and II ovarian cancer are alive without recurrence 1.8-9.8 years (median, 4.5 years) after diagnosis. Two of the three Stage III patients have died of disease: one at 4.3 years and one at 7.7 years after detection. Four patients developed ovarian cancer within 12 months of a negative scan (FN): 2 Stage II, 2 Stage III. Three of these patients are alive with no evidence of disease 0.4, 1.9, and 5.5 years after diagnosis, and 1 patient has died of disease 0.7 years after diagnosis. Four patients developed ovarian cancer more than 12 months following a normal screen. All 4 presented clinically with Stage III disease. Two of these patients have died of disease and two patients are alive 1.5 and 2.1 years after diagnosis. TVS screening was associated with the following statistical variables: sensitivity, 81%; specificity, 98.9%; positive predictive value (PPV), 9.4%; and negative predictive value (NPV), 99.97%. After 46, 113 screening years, there have been 3 ovarian cancer deaths in the annually screened population and 2 ovarian cancer deaths in women receiving less than annual screening. The survival of ovarian cancer patients in the annually screened population was 95.0 +/- 4.9% at 2 years and 88.2 +/- 8.0% at 5 years. Excluding all cases of nonepithelial or borderline epithelial malignancies, the survival of patients with ovarian cancer in the annually screened population was 92.9 +/- 6.9% at 2 years and 83.6 +/- 10.8% at 5 years. CONCLUSIONS: (1) TVS screening, when performed annually, is associated with a decrease in stage at detection and a decrease in case-specific ovarian cancer mortality. (2) TVS screening does not appear to be effective in detecting ovarian cancer in which ovarian volume is normal. PMID- 10831342 TI - Information needs and decisional preferences among women with ovarian cancer. AB - OBJECTIVES: Studies show that women with cancer want more information about and participation in all aspects of their healthcare including decision-making. However, most studies have been done on women with breast cancer, which often runs a lengthy course and has strong patient-advocacy groups. Little is known about the preferences of women with ovarian cancer, the fifth leading cause of cancer death in women, which often has a more rapidly fatal course. METHODS: This study of women with ovarian cancer investigates what information is most vital for women with ovarian cancer, their preferred role in decision-making, and the influence of sociodemographic, disease-related, and psychological factors. RESULTS: Questionnaires were completed by 105 women with ovarian cancer in two Canadian university hospital oncology clinics. Their mean age was 55.8 +/- 14. 9 years. Most were married, well-educated, and considered their health status to be excellent or good, even though over 60% had metastatic disease. Over 80% of these women wanted detailed information about ovarian cancer during the diagnosis, treatment, and posttreatment stages of their disease. In particular, they wanted information pertaining to the disease, treatment, and self-care issues. Approximately 60% of women preferred to share decision-making with their doctors at every stage of the illness. Psychological variables and disease severity were found to influence information needs and decisional preferences. In general, the more psychologically distressed the women, the more information they wanted about coping strategies and the more serious the illness, the more shared decision making was desired. CONCLUSION: These results present a challenge to health care providers in more adequately meeting the individual information needs of women with ovarian cancer and involving them to the extent that they wish in the decision-making process. PMID- 10831343 TI - Splenectomy and surgical cytoreduction for ovarian cancer. AB - OBJECTIVE: The aim of this study was to evaluate the role of splenectomy, as a surrogate marker for aggressive tumor cytoreduction in ovarian cancer, and its impact on patient morbidity and survival. METHODS: A retrospective cohort study of 35 patients who underwent splenectomy for ovarian cancer cytoreduction between August 1986 and May 1998 was performed. Data abstracted from the medical record included patient demographics, preoperative imaging, surgical procedures, tumor distribution, postoperative complications, chemotherapy treatment, and follow-up information. RESULTS: Splenectomy was performed in 13 patients at the time of primary cytoreduction and in 22 patients at the time of secondary cytoreduction. Preoperative diagnosis of splenic involvement was frequently made prior to secondary surgery, 77.3% compared to 15.4% of primary cases. In addition, parenchymal splenic involvement was more commonly observed at recurrence, 59.1% vs 23.1% at initial presentation. Disease distribution in secondary cytoreduction cases tended to be more focal, macronodular, and have no ascites. Cytoreduction to less than 1 cm disease was achieved in 100% of primary patients and 86% of secondary patients. Major morbidity (pneumonia, PE, sepsis, pancreatitis, MI) occurred in 23.1% of primary patients and 28.6% of secondary patients. Combining splenectomy with other cytoreductive procedures may make splenectomy itself seem more morbid. With a 17-month median follow-up, median progression-free interval was 24 months in primary patients and 14 months in secondary patients. Among secondary patients, median survival time after splenectomy and cytoreduction was 41 months. CONCLUSIONS: Splenectomy at the time of primary and secondary cytoreduction for ovarian cancer can be performed with acceptable morbidity. Secondary cytoreduction patients may be selected preoperatively by their progression-free interval, prior degree of cytoreduction, and macronodular tumor involvement on imaging studies. Identification of splenic involvement allows for appropriate counseling and preoperative preparation. PMID- 10831344 TI - Differential expression of matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 protein and mRNA in epithelial ovarian tumors. AB - OBJECTIVE: Matrix metalloproteinase-9 (MMP-9) can degrade gelatin and type IV collagen and is known to play an important role in tumor cell invasion across the basement membrane. The tissue inhibitor of metalloproteinase-1 (TIMP-1) is able to prevent activation of pro-MMP-9 and forms a 1:1 complex with the active form of MMP-9. The aim of the present study was to investigate the expression of MMP-9 and TIMP-1 in benign, borderline, and invasive epithelial ovarian tumors. MATERIALS AND METHODS: A total of 90 patients with epithelial ovarian tumor were treated at the Brigham and Women's Hospital and were used as the study population. Immunohistochemistry and in situ hybridization were performed to detect protein and mRNA expression of MMP-9 and TIMP-1. RESULTS: In the 90 epithelial ovarian tumors tested, MMP-9 expression in tumor cells was found to be significantly enhanced in serous and mucinous ovarian carcinomas compared with benign and borderline tumors. We also observed the immunostaining of MMP-9 in stromal cells of benign, borderline, and invasive epithelial ovarian tumors. Moreover, the expression levels of TIMP-1 in tumor cells were significantly higher in borderline and invasive ovarian tumors than in benign tumors. CONCLUSION: Using an in situ hybridization technique, we disclosed a direct correlation between the presence of mRNA and protein expression for both MMP-9 and TIMP-1. The present data suggest that high levels of MMP-9 protein in invasive epithelial ovarian carcinoma are strongly associated with tumor cell invasion. Enhanced expression of TIMP-1 protein in borderline and invasive tumors indicates that endogenous TIMP-1 protein may play a paradoxical role in ovarian tumor progression. PMID- 10831345 TI - Flt-3 ligand inhibits growth of human ovarian tumors engrafted in severe combined immunodeficient mice. AB - OBJECTIVE: The current study evaluated the effects of Flt-3 ligand (FL) on the growth of human malignant ovarian tumors engrafted in severe combined immunodeficient (SCID) mice with particular attention directed at FL's effect on the host natural killer (NK) cell response against ovarian cancer xenografts. METHODS: Equal portions of surgical specimen-derived human ovarian carcinomas were engrafted subcutaneously (SC) into SCID mice. Mice were placed into one of two treatment groups 7 days after the day of implantation. Group 1 received placebo injections SC from Day 1 to Day 20 and group 2 received FL at 10 microg/day SC from Day 1 to Day 20. NK cell depletion was performed on three additional mice from group 2 starting on Day 0 using anti-asialo GM1. Serial tumor volumes were measured. On Day 21, mice from each group were sacrificed, and tumors and spleens were evaluated. Data analysis included chi(2) tests, Student t tests, and analyses of variance when appropriate. RESULTS: FL resulted in tumor growth delay compared with control (P = 0.036). When NK cell activity was depleted prior to FL administration, no tumor growth delay was observed. Spleens from FL-treated mice were larger (P < 0.01) with expanded white pulp compared with controls. Histologic examination of tumor sections from FL-treated mice revealed regions of solid tumor growth with glandular architecture similar to that seen in control tumors; however, there was an obvious increase in regions composed largely of dense fibrosis in the FL-treated tumors. NK cells and other infiltrating cells could be detected in clusters among tumors from mice treated with FL whereas these cells were only occasionally detected in sections of control tumors. CONCLUSION: FL treatment resulted in an antitumor response against human ovarian cancer engrafted in SCID mice and this inhibition appears to be largely host NK cell mediated. The tumor inhibition seen in this model is similar to that previously seen using syngeneic tumors grown in an immunocompetent animal model. Results from this model can potentially be extrapolated to treatment of human ovarian cancer patients. PMID- 10831346 TI - Effect of topotecan on serum CA-125 in patients with advanced epithelial ovarian cancer. AB - The effect of topotecan on CA-125 serum levels was evaluated in 30 patients with advanced epithelial ovarian cancer. All patients had progressive disease and were relapsing during (11 patients) or after (19 patients) chemotherapy containing paclitaxel and platinum. Topotecan (1.0 mg/m(2)/day) was administered intravenously on Days 1-5 every 3 weeks. The patients had received a median of 2 (1-5) prior regimens. Four patients had increased CA-125 only, and 26 had both measurable disease and increased CA-125. Two patients (7%) achieved a clinical partial response with durations of 5 and 10+ months, respectively. Eighteen other patients (60%) exhibited no clinical change with a median duration of 5+ months (range: 2-11+ months). Among these patients 9 (30%) had a biochemical response. The rate of change in CA-125 (s, slope of the exponential regression curve) during treatment with topotecan was compared with s over a period before treatment. A decrease in s was observed in 20 patients (74%). Comparing the mean values of s before and during topotecan, a significant (P = 0.005) decrease was seen in the CA-125 serum levels. The mean doubling times before and during treatment were 59 and 1421 days, respectively. Toxicity was mainly hematologic. Neutropenia grades III and IV were seen in 16 and 10 patients, respectively. No patients died due to side effects. Generally the side effects were mild to moderate. In conclusion, at the given dose intensity topotecan shows activity in advanced paclitaxel- and platinum-resistant ovarian cancer based on CA-125 measurements. PMID- 10831347 TI - Expression of epidermal growth factor receptor-related family products in gestational trophoblastic diseases and normal placenta and its relationship with development of postmolar tumor. AB - OBJECTIVE: The goal of this work was to study the expression of epidermal growth factor receptor (EGFR) and c-erbB-3 and c-erbB-4 oncogenes in gestational trophoblastic diseases and normal first-trimester placenta. STUDY DESIGN: Paraffin sections of 16 cases of partial mole, 25 cases of complete mole, 10 cases of gestational choriocarcinoma, and 11 cases of therapeutic abortion were studied immunohistochemically for EGFR, c-erbB-3, and c-erbB-4 proteins. The presence of EGFR mRNA was studied using in situ hybridization. RESULTS: Staining for EGFR was detected immunohistochemically in all cell types in gestational trophoblastic diseases and normal placenta. In situ hybridization for EGFR mRNA correlated with immunostaining for EGFR in all tissues studied. All 10 cases of choriocarcinoma exhibited strong immunoreactivity for EGFR. The levels of expression of EGFR in choriocarcinoma and syncytiotrophoblasts and cytotrophoblasts in complete mole were significantly greater than those in syncytiotrophoblasts and cytotrophoblasts in both normal placenta and partial mole (P < 0.01, P < 0.01). Expression of c-erbB-3 did not significantly differ among placental and gestational trophoblastic disease tissues and trophoblastic cell types except for significantly increased expression in choriocarcinoma as compared with cytotrophoblasts of partial mole (P = 0.02). The placenta, complete and partial mole, and choriocarcinoma tissues demonstrated similar immunoreactivity for c-erbB-4. Strong immunostaining for EGFR (P = 0.02) and c erbB-3 (P < 0.01) in extravillous trophoblasts of complete mole was found to be significantly correlated with the development of persistent postmolar gestational trophoblastic tumor. CONCLUSION: The EGFR-related family of oncogenes may be important in the pathogenesis of gestational trophoblastic diseases. The increased expression of EGFR and c-erbB-3 in complete mole may also influence the development of persistent gestational trophoblastic tumor. PMID- 10831348 TI - Telomerase activity in Papanicolaou smear-negative exfoliated cervical cells and its association with lesions and oncogenic human papillomaviruses. AB - OBJECTIVE: The goal of this study was to evaluate telomerase activity in exfoliated cervical cells and its association with cytology, pathology, and human papillomavirus (HPV). METHODS: Telomerase activity and HPV DNA sequences were examined in the exfoliated cervical cells from a general population of 245 women aged more than 30 years undergoing routine cervical screening by Papanicolaou smear. The women who were found to have telomerase activity or abnormal cytology in their exfoliated cervical cells were examined for cervical lesions by colposcopy and biopsy. RESULTS: Cytology for our population (mean, 56 years) revealed only one abnormal smear (1/245, 0.4%), in which a cervical intraepithelial neoplasia grade I (CIN I) lesion was found. The exfoliated cervical cells used to prepare the smear were negative for telomerase and contained low-risk HPV DNA. Telomerase activity was found in 16 exfoliated cell samples (16/245, 6.5%); high-risk HPV DNA was found in 9 of these samples (9/16, 56%) and 9 of the biopsy specimens that could be evaluated from patients testing positive for telomerase revealed CIN I lesions (9/11, 82%). CONCLUSIONS: Telomerase activity is often associated with high-risk HPV infection and it is suggested that telomerase assay can help to detect occult cervical lesions. PMID- 10831349 TI - Detection of micrometastasis by cytokeratin-20 (reverse transcription polymerase chain reaction) in lymph nodes of patients with endometrial cancer. AB - BACKGROUND: Cytokeratins are constituents of the intermediate filaments (IFs) of epithelial cells which are expressed in various combinations, depending on the type of epithelium and degree of differentiation. We have reported (R. Zemer, A. Fishman, J. Bernheim, S. Zimlichman, O. Markowitz, M. Altaras, and A. Klein, Gynecol Oncol 70:410-413, 1998) on the determination of cytokeratin-20 (CK-20) by reverse transcription polymerase chain reaction (RT-PCR) in the detection of endometrial cancer cells as a potential biomarker. In that study, we also found that by using immunocytochemistry, most carcinomas were found to be negative for CK-20. The sensitivity and specificity rates obtained by using the RT-PCR method were 94.4 and 91%, respectively. OBJECTIVE: The aim of this study is to investigate the feasibility and potential of the specific mRNA marker, CK-20, to detect endometrial cancer cells-micrometastases (MMs)-by RT-PCR in lymph node (LN) samplings of patients undergoing hysterectomy for endometrial carcinoma. METHOD: We used the RT-PCR method to determine the expression of CK-20 in the LNs of 20 patients [study group (SG)] who were being surgically staged and treated for endometrial carcinoma. The specificity of the mRNA CK-20 marker was examined in LNs obtained from five healthy patients [control group (CG)] who underwent abdominal hysterectomy and bilateral salpingo-oopherectomy for benign gynecologic conditions. The LNs obtained from the SG and CG patients were prepared together before mRNA extraction. RNA of the various cell pellets was extracted and RT-PCR was performed with CK-20 primers. RT-PCR products were analyzed by agarose gel electrophoresis and ethidium bromide staining against PCR size markers. Specificity of the RT-PCR products was examined by Southern blotting. RESULTS: Histopathologic examinations demonstrated the presence of metastases in two (10%) SG patients. These patients were also CK-20 positive. Of the remaining 18 patients with negative histopathologic results, 6 (33%) were CK-20 positive and 12 (67%) were negative. All the CG patients were CK-20 negative (specificity, 100%). CONCLUSIONS: The results obtained in this study suggest that RT-PCR of CK 20 is more sensitive than traditional histopathologic methods in the diagnosis of MMs in LNs of patients with endometrial cancer. Thus, due to the aforementioned characteristics of CK-20, it may be considered a powerful biomarker in the detection of MMs in LNs of patients with endometrial cancer. PMID- 10831350 TI - Clinical significance of combined use of macrophage colony-stimulating factor and squamous cell carcinoma antigen as a selective diagnostic marker for squamous cell carcinoma arising in mature cystic teratoma of the ovary. AB - OBJECTIVES: Mature cystic teratoma of the ovary transforms into malignant tumors, mostly squamous cell carcinomas, at an incidence of approximately 2%. Preoperative diagnosis of squamous cell carcinoma arising in mature cystic teratoma of the ovary is a difficult task. The present study aims to assess whether combined use of two serum tumor markers, macrophage colony-stimulating factor (M-CSF) and squamous cell carcinoma antigen (SCC), is effective in preoperatively diagnosing squamous cell carcinoma arising in mature cystic teratoma of the ovary, distinguishing it from mature cystic teratoma without malignant transformation. METHODS: Serum levels of M-CSF and SCC were assayed using blood samples collected preoperatively from 31 patients with squamous cell carcinoma arising in mature cystic teratoma of the ovary and 133 patients with mature cystic teratoma of the ovary without malignant transformation. RESULTS: In 22 of the 31 (71.0%) patients with squamous cell carcinoma arising in mature cystic teratoma of the ovary, the serum M-CSF levels exceeded the upper limit of the normal level (1056 U/ml). This positive incidence of the elevated serum M-CSF levels was significantly higher compared with that (13.5%, 18/133) observed in patients with benign cystic teratoma of the ovary (P < 0.0001). Regarding the serum levels of SCC, 13 of 31 (41.9%) patients with malignant tumors showed positive values exceeding the cutoff value of 2.0 ng/ml. Again, this incidence of positive cases was significantly higher compared with that (15.0%, 20/133) observed in patients with benign tumors (P < 0.01). There was no correlation between the serum levels of M-CSF and SCC among patients with squamous cell carcinoma arising in mature cystic teratoma of the ovary. Patients with malignant tumors testing positive for elevated M-CSF did not necessarily test positive for SCC. Patients with positive values for excess M-CSF and/or SCC constituted 87.1% of the total (27/31). Even when patients were restricted to those with stage I tumors, a value as high as 83.3% (15/18) was still obtained for those testing positive for elevated M-CSF and/or SCC. CONCLUSION: Serum M-CSF was proven to be useful as a tumor marker for detecting squamous cell carcinoma arising in mature cystic teratoma of the ovary. Combined use of serum M-CSF and SCC as a marker seemed to be useful in the selective diagnosis of mature cystic teratoma of the ovary harboring malignant squamous carcinoma, discriminating it from that without malignant carcinoma. PMID- 10831351 TI - Ovarian volume related to age. AB - OBJECTIVE: The goal of this study was to determine the relationship of ovarian volume to age, height, and weight in women undergoing transvaginal sonography. METHODS: Thirteen thousand nine hundred sixty-three women 25-91 years of age undergoing annual transvaginal sonography as part of the University of Kentucky Ovarian Cancer Screening Program were the subjects for this investigation. Each ovary was measured in three dimensions, and ovarian volume was calculated using the prolate ellipsoid formula (L x H x W x 0.523). Mean ovarian volume according to age was calculated for each decade of life. RESULTS: Data were obtained from 58,673 observations of ovarian volume. Mean ovarian volume was 6.6 +/- 0.19 cm(3) in women less than 30 years of age; 6.1 +/- 0.06 cm(3) in women 30-39; 4.8 +/- 0.03 cm(3) in women 40-49; 2.6 +/- 0.01 cm(3) in women 50-59; 2. 1 +/- 0.01 cm(3) in women 60-69; and 1.8 +/- 0.08 cm(3) in women >/=70. Mean ovarian volume was 4.9 +/- 0.03 cm(3) in premenopausal women and 2.2 +/- 0.01 cm(3) in postmenopausal women (P < 0.001). The use of exogenous estrogens was associated with a significant reduction in ovarian volume in women 40-59 years of age, but not in women >/= 60. Ovarian volume was unrelated to patient weight but was greater in tall women (>68 in.) than in short women (<58 in.). CONCLUSION: There is a statistically significant decrease in ovarian volume with each decade of life from age 30 to age 70. Mean ovarian volume in premenopausal women is significantly greater than that in postmenopausal women. The upper limit of normal for ovarian volume is 20 cm(3) in premenopausal women and 10 cm(3) in postmenopausal women. PMID- 10831352 TI - Prognostic significance of vascular endothelial growth factor and its receptors in endometrial carcinoma. AB - OBJECTIVE: The aim of this study has been to evaluate the clinical significance of expression of VEGF and its receptors, Flt-1, KDR, and Flt-4, in endometrial carcinomas. METHODS: Specimens of endometrial carcinomas from 86 patients were investigated immunohistochemically using respective specific antibodies. Additionally, samples from 14 patients with complex atypical endometrial hyperplasia (AEH) and 15 patients with normal endometria were also examined. Immunohistochemical assessment was classified as negative, weakly positive, and strongly positive according to staining intensity and the percentage of positive cells. RESULTS: In positive cases, VEGF and its receptors were usually expressed homogeneously in the cytoplasms of cells in the endometrial carcimona, similar to the staining intensity of endothelial cells of stromal microvessels adjacent to carcinoma nests. The overall positive rates in the 86 carcinoma specimens were 66% for VEGF, 51% for Flt-1, 38% for KDR, and 57% for Flt-4. Their expressions in endometrial carcinoma tissues were high with significance or with borderline significance, compared to those in samples of complex AEH or normal endometria. Survival curves determined by the Kaplan-Meier method and univariate analysis showed VEGF, Flt-1, and Flt-4 overexpression to be related to poor prognosis of patients with endometrial carcinomas. However, multivariate analysis revealed that Flt-4 overexpression correlated independently with poor survival, similar to a value for myometrial invasion at one-half or more and that for retroperitoneal lymph node metastasis, whereas VEGF and Flt-1 overexpression did not. CONCLUSION: Flt-4 overexpression might be a promising prognostic indicator for survival of a patient with endometrial carcinoma. PMID- 10831353 TI - Differential expression of antiapoptotic genes in human endometrial carcinoma: bcl-XL succeeds bcl-2 function in neoplastic cells. AB - OBJECTIVES: Previous histochemical observations have suggested a possible involvement of the bcl-2 family genes in the acquisition of neoplastic phenotype of the endometrium. Since knowledge of the type and function of genes controlling the transformed cell may result in new diagnostic, prognostic, and therapeutic approaches, we have investigated at the molecular level the biological role of bcl-2 family genes in endometrial neoplastic cells. METHODS: To investigate the relationship between the sensitivity to apoptosis and the expression of the bcl-2 family genes, we set up a model system consisting of four human endometrial carcinoma cell lines. This system constitutes an array of two cell pairs presenting, respectively, endometrioid and adenosquamous phenotypes. G2 and G3 gradings are represented within each pair; in addition, each set contains one cell line that is apoptosis-sensitive and one that is resistant. Transfection of bcl-2 and bcl-XL into apoptosis-sensitive cells was used to monitor the biological function of protective genes. RESULTS: A differential pattern of expression of bcl-2 family genes was observed in apoptosis-sensitive versus resistant cells, independent from the histological subtype. Resistant lines exhibited high amounts of Bcl-XL and low amounts of Bcl-2. Bax expression clearly correlates with cellular susceptibility to apoptosis. Transfection of bcl-XL resulted in a dose-dependent enhancement in resistance toward apoptosis. In contrast, the main effect of bcl-2 constitutive overexpression was to drastically abate the proliferative potential of transfected cells. CONCLUSIONS: These data demonstrate, at the molecular level, that bcl-XL is selected as an apoptosis protective gene in place of bcl-2 while bax retains its dominant proapototic role. PMID- 10831354 TI - Phase II study of mifepristone (RU486) in refractory ovarian cancer. AB - OBJECTIVE: A phase II study of Mifepristone (RU486) was conducted in patients with ovarian cancer whose tumors were resistant to cisplatin and paclitaxel, alone or in combination. PATIENTS AND METHODS: Forty-four patients were accrued into this study. All had ovarian cancer that had become resistant to cisplatin and paclitaxel. Patients received Mifepristone 200 mg orally on a daily basis. Patients were followed by tumor size or CA-125 levels when there was no measurable disease. A dose reduction of Mifepristone was to occur in the event of grade 3/4 hematologic, GI, or liver toxicity, creatinine >2.5%, and grade 4 peripheral neuropathy. RESULTS: Thirty-four patients were evaluable for response. Nine (26.5%) of these patients had a response to Mifepristone. Three(9%) patients had a complete response, and six (17.5%), a partial response. The response of one patient in each group was measured by CA-125 levels while the remainder had measurable disease. The response lasted 1 to 4 months in all but one patient. One patient continues to respond after more than 3 years. The major toxic effect was a rash and this was the major reason patients were removed from the study. CONCLUSION: Mifepristone has activity against ovarian cancer resistant to cisplatin and paclitaxel. The drug is well tolerated. Further studies need to be performed when this drug becomes more widely available in the United States. PMID- 10831355 TI - Phase II trial of combination intraperitoneal cisplatin and 5-fluorouracil in previously treated patients with advanced ovarian cancer: long-term follow-up. AB - OBJECTIVES: This trial was performed to determine the response rate and progression-free and overall survivals of patients with advanced recurrent ovarian cancer who were treated with intraperitoneal cisplatin and 5 fluorouracil. METHODS: Twenty-four patients with ovarian cancer were entered on this trial and treated with intraperitoneal (ip) cisplatin (DDP) and ip 5 fluorouracil, every 3 weeks for eight cycles. Following iv hydration, the cisplatin and 5-fluorouracil were administered through an ip catheter in 2 liters of 0.9% normal saline with a 4-h dwell. RESULTS: All patients were evaluable for progression-free and overall survival and toxicity analysis, and 22 patients for response. The median age was 59 (range, 35-71); initial disease status included 9 patients with residual disease following chemotherapy prior to entry on this study; 5 patients had progressed, and 10 patients had recurrent disease more than 6 months following initial chemotherapy. Of the 9 patients with residual disease, 1 complete response and 3 partial responses were observed; of 10 patients with recurrent disease, 1 complete and 1 partial response were observed for an overall response rate of 27%. No objective responses were seen in the 7 patients who were platinum-refractory on protocol entry. The median progression-free and overall survivals are 7.0 (range, 0.5-137) and 15.5 (range, 3-147) months, respectively. Toxicity included hypomagnesemia, vomiting, abdominal pain, and mild anemia. Only one patient required a dosage adjustment of cisplatin for a serum creatinine elevation >2.0 mg/dl. CONCLUSIONS: We conclude that the combination of ip cisplatin and 5-FU is an effective regimen for patients with residual or relapsed epithelial ovarian cancer with survival durations, response rates, and toxicity profiles that compare favorably with those of other second-line ovarian cancer regimens. Patients who are primarily platinum-refractory are unlikely to benefit from these agents administered into the peritoneal cavity. PMID- 10831356 TI - p27 and cyclin D1 abnormalities in uterine papillary serous carcinoma. AB - OBJECTIVE: The expression status of p27 and cyclin D1 was examined in 21 uterine papillary serous carcinoma (UPSC) specimens to determine the role of these genes in the development of this disease. The status of p53, p16, Rb, and K-ras was also determined in these tissues so that a marker profile for UPSC could be compared with the published marker profile for other forms of endometrial and ovarian cancer. METHODS: Immunohistochemistry was performed on 21 UPSC tissue sections to determine the expression status of p27, cyclin D1, p53, p16, and Rb. K-ras mutations were identified by restriction fragment length polymorphism analysis of DNA isolated from the UPSC sections. RESULTS: All specimens displayed at least one molecular abnormality. A high incidence of p27 alterations were observed, with reduced p27 expression measured in 16 of 21 (76%) tumors, followed by p53 alterations observed in 13 of 21 (62%) tumors. The p27 abnormalities occur at an early stage of the disease, with 63% (5/8) of Stage I cases displaying reduced p27 expression. Cyclin D1 overexpression was observed in 4 of 21 (19%) specimens, whereas p16, Rb, and K-ras abnormalities were each observed in 2 of 21 specimens (10%). Both K-ras mutations were at codon 12. The p16 and Rb abnormalities coexisted in the same specimens. CONCLUSION: UPSC tumors display a high incidence of p27 abnormalities, suggesting that p27 abnormalities play an important role in the development of this disease. Our results also indicate that cyclin D1 overexpression is involved in the development of a small number of UPSC cases. A comparison of our results with reports by other authors suggests that UPSC shares molecular marker alterations with both ovarian cancer and endometrioid adenocarcinoma. PMID- 10831357 TI - Topotecan in squamous cell carcinoma of the cervix: A Phase II study of the Gynecologic Oncology Group. AB - PURPOSE: The activity and toxicity of topotecan were evaluated in a multicenter Phase II study for patients with previously treated squamous cell carcinoma of the uterine cervix. PATIENTS AND METHODS: Histologic confirmation of the primary diagnosis was required, as well as adequate performance status and vital organ function and the presence of measurable disease. Patients were allowed one prior regimen of systemic therapy, usually platinum-based. A two-stage accrual design was utilized with early stopping criteria and monitoring of toxicity. Topotecan was administered at 1.5 mg/m(2) per day for 5 consecutive days on a 21-day cycle with modifications based on hematologic toxicity. RESULTS: Forty-five patients were entered. Two patients were ineligible (incorrect tumor type) and 2 were inevaluable (never received therapy). One additional patient was not evaluable for response (nonmeasurable disease). A median of 2 cycles was administered to each patient (range: 1-17 cycles) with grade 4 neutropenia in 68% and grade 4 thrombocytopenia in 39% of patients, but without treatment-related deaths. Nonhematologic toxicity was generally mild and not dose-limiting. The overall (complete and partial) response rate among evaluable patients with measurable disease was 12.5% with stable disease in an additional 37. 5%. Median progression free survival was 2.1 months. CONCLUSIONS: As a single agent topotecan shows modest antitumor activity, with manageable hematologic and nonhematologic toxicity, in patients with previously treated squamous cell carcinoma of the cervix. Further evaluation in chemotherapy-naive patients or in combination with cisplatin and/or radiation may be indicated. PMID- 10831358 TI - National Cancer Data Base/Surveillance Epidemiology and End Results: potential insensitive-measure bias. AB - OBJECTIVES: Abstraction of data from National Cancer Data Base (NCDB)/Surveillance Epidemiology and End Results (SEER) for reasons other than incidence, mortality, and patterns of care has risen. A potential problem with these data is that insensitive-measure bias can exist because of possible inaccuracies in hospital tumor registry staging. The purpose of this study is to assess the accuracy of tumor registry staging from six community hospitals. METHODS: Staging of 103 consecutive cancers operated on by a gynecologic oncologist (one of the authors) as a surgical consultant to a gynecologist or surgeon was reviewed. Hospital tumor registry staging forms were arbitrarily assigned to be completed by the nongynecologic oncologist versus the gynecologic oncologist by the medical records department. The authors reassessed cancer staging by medical chart review. The tumor registry staging was compared with the actual staging as determined by the authors. Major staging violations were defined as errors that would significantly change stage enough to alter prognosis or change recommended adjuvant treatment. All other violations were defined as minor. RESULTS: Twenty-eight (27%) cancers were staged by the gynecologic oncologist and 75 (73%) by nongynecologic oncologists. Eighty (78%) cancers were endometrial and 14 (13%) ovarian. Eighty-three (81%) tumors were stage I or II. Major staging violations occurred in 0% of cancers staged by the gynecologic oncologist and 22% (16/75) by a nongynecologic oncologist (P = 0.002). Minor staging violations occurred in 14% (4/28) of cancers staged by the gynecologic oncologist and 42% (32/75) by a nongynecologic oncologist (P = 0. 005). Minor violations were due to omission of histologic subtype and/or grade. CONCLUSION: The 22% major staging violation rate represents significant insensitive-measure bias. If additional studies produce similar results, abstraction of data from NCDB/SEER for reasons other than incidence, mortality, and patterns of care cannot be accepted as evidence-based scientific medicine. PMID- 10831359 TI - Classification of asymptomatic adnexal masses by ultrasound, magnetic resonance imaging, and positron emission tomography. AB - OBJECTIVE: The purpose of this study was to determine the diagnostic accuracy of sonography versus magnetic resonance imaging (MRI) and positron emission tomography (PET) in the characterization of adnexal masses. METHODS: One hundred and one patients with asymptomatic adnexal masses, which were scheduled for laparoscopy, underwent preoperative transvaginal ultrasound, MRI, and 2 [(18)F]fluoro-2-deoxy-d-glucose PET. Two different sonomorphological scoring systems were used to distinguish malignant from benign lesions. In addition, transvaginal Doppler flow velocimetry was performed and the resistance index (RI) of ovarian blood vessels was calculated. RI values below 0.45 were considered to indicate malignancy. MRI was evaluated on the basis of signal intensity and morphologic features such as wall thickness, septations, fluid or solid components, and vascularity. PET imaging was used to determine 2-[(18)F]fluoro-2 deoxy-D-glucose uptake. Malignancy was suspected if radiotracer uptake equaled or exceeded that of the liver. Based on histologic findings, sensitivity, specificity, positive and negative predictive values, and accuracy were first calculated independently for each imaging technique. Finally, a second session resulted in a consensus diagnosis being made based on the findings of all three modalities. RESULTS: Sonographic evaluation of adnexal masses resulted in correct classification of 11 of 12 ovarian malignancies (sensitivity 92%) but with a specificity of only 60%. With MRI and PET, specificities improved to 84 and 80% respectively, but sensitivities decreased. When all imaging modalities were combined, sensitivity and specificity were 92 and 85%, respectively, and accuracy was 86%. CONCLUSION: Combination of ultrasound with MRI and PET may improve accuracy in differentiation of benign from malignant ovarian lesions. However, negative MRI or PET results do not rule out early-stage ovarian cancer or borderline malignancies. PMID- 10831360 TI - Human immunodeficiency virus infection and invasive cervical cancer in South Africa. AB - PURPOSE: The goal of this study was to determine whether South African HIV seropositive women with invasive cervical cancer present with disease that is more advanced than that of HIV-seronegative women and whether degree of immunosuppression affects the extent of disease at initial presentation. METHODS: This study is a retrospective review of 60 HIV-seropositive and 776 HIV seronegative new cases of invasive cervical carcinoma seen at the combined gynecologic oncology unit of the University of the Witwatersrand, Johannesburg, South Africa. RESULTS: The HIV seroprevalence was 7.2%. Squamous cell carcinoma was the histologic subtype in more than 90% of both cohorts of patients. Although the HIV-positive patients presented with invasive cervical cancer almost 10 years earlier than the HIV-negative patients, i.e., mean age 44 years +/- 9.8 versus 53 years +/- 12.7, respectively (P R(2)) is [x, y]mapsto[log(k)(x), log(k)(y)]. The solution is generalized to arbitrarily many dimensions by substituting the sin and cos in the generalized Weber law by the standard coordinates of a unit vector. Copyright 2000 Academic Press. PMID- 10831376 TI - TELEGRAPHIC REVIEWS. PMID- 10831377 TI - Taenia solium: a cysteine protease secreted by metacestodes depletes human CD4 lymphocytes in vitro. AB - Excreted/secreted products from Taenia solium metacestodes cultured in vitro were analyzed for peptidase activity using peptide substrates Z-Phe-Arg-AFC, Arg-AFC, and Z-Gly-Gly-Arg-AFC and zymography studies. Specific inhibitor profiles revealed mainly cysteine and metalloprotease activities. Hydrolysis of substrate Z-Phe-Arg-AFC was augmented by the addition of L-cysteine and acid pH, consistent with cysteine protease activity. Cysteine protease activity was more prominent in supernatants from living metacestodes cultured in PBS than in either RPMI or RPMI plus fetal calf serum and was proportional to the number of metacestodes. Flow cytometry analysis showed depletion of human T lymphocytes cultured with living T. solium metacestodes. CD4(+) expression was significantly decreased when metacestode E/S products and L-cysteine were added to lymphocyte cultures (P = 0.027). This peptidase activity was inhibited by E-64 indicating that the depletion of CD4(+) cells was due to cysteine protease activity. Thus, T. solium metacestodes produce excretory/secretory proteases. These enzymes may cleave molecules critical for the host immune response allowing the parasites to survive in the host tissues. PMID- 10831378 TI - Leishmania (Viannia) guyanensis: a new minicircle class exclusive to this specie isolated from a DNA cosmid library useful for taxonomic purposes. AB - A new minicircle class exclusive to this specie isolated from a DNA cosmid library useful for taxonomic purposes. Experimental Parasitology 94, 143-149. In this paper we describe a new minicircle class exclusive to Leishmania (Viannia) guyanensis. The minicircle class was obtained with the aid of a total DNA cosmid library. The library was screened with an EcoRI fragment isolated from L. (V.) guyanensis (M4147). After screening seven clones were selected which showed strong hybridisation. Clones were digested and hybridised with the same probe. After hybridisation only one clone containing the desired fragment was positive. The fragment sized around 1000 bp was subcloned into pBluescript for sequencing. Sequence analysis using the GCG programme showed no substantial homology with any sequences previously reported, apart from the expected homology with the conserved region of Leishmania kDNA sequences. The probe hybridised strongly only to L. (V.) guyanensis kDNA after medium stringency washing. PMID- 10831379 TI - Interleukin 1alpha activity of peritoneal and bone marrow macrophages infected with Leishmania major and Leishmania donovani in vitro. AB - In this study, the pattern of interleukin-1alpha (IL-1alpha) production by both peritoneal (PM) and bone marrow macrophages (BMM) from resistant (C3H/HeJ) and susceptible (BALB/c) mice was investigated, using a bioassay and an IL-1alpha specific ELISA kit. PM from normal uninfected mice showed either an initial high (C3H/HeJ) or a neglected (BALB/c) level of IL-1alpha activity, respectively, probably due to thioglycollate stimulation. Infection with Leishmania major induced only a marginal effect on IL-1 production by both cells. Normal, uninfected and unstimulated BMM from both mice did not produce IL-1alpha over a 7 day period of cultivation in vitro. Upon stimulation with either lipopolysaccharide (LPS) (BALB/c) or concanavalin A (Con A) (C3H/HeJ), both cell types produced IL-1alpha that peaked within the first 12-24 h following stimulation. BMM from C3H/HeJ and BALB/c mice failed to produce IL-1alpha when infected in vitro with L. major or L. donovani promastigotes. However, infection with these two parasites did not interfere with the capability of the host cell to produce IL-1alpha when stimulated with LPS or Con A. The level of IL-1alpha production was independent of the degree of parasitization of the macrophages. Similar results were observed with IL-1beta and IL-6 production by BMM, even though their levels were generally slightly higher than those obtained with IL 1alpha. PMID- 10831380 TI - Onchocerca volvulus: comparative analysis of antibody responses to recombinant antigens in two animal models of onchocerciasis. AB - Experimental infections of chimpanzees with Onchocerca volvulus and cattle with Onchocerca ochengi provide model systems for research in human onchocerciasis. These infections share many similarities from the standpoint of parasite biology, but little is known about the comparability of immune responses in the two systems. To make a direct comparison between the models in terms of immune responsiveness to defined parasite products, three recombinant antigens of O. volvulus (Ov7, Ov103, and B20) were used to analyze the kinetics of antibody production following experimental infection. Each of the antigens was derived from adult cDNA libraries following immunoscreening with sera from chimpanzees (Ov7, Ov103) or cattle (B20). All chimpanzees (n = 12) and cattle (n = 8) displayed responses to Ov7 and Ov103, and all cattle, but only 33% of chimpanzees, showed responses to B20. The dynamics of the response to individual antigens showed further similarities between the chimpanzees and the cattle, with responses to Ov7 and Ov103 peaking after, and B20 before, the onset of patent infections. We conclude that there is good preliminary evidence of concordance in the kinetics of serological responses in the two models. However, individual antigens many be more or less immunogenic in the two systems, making it inadvisable to extrapolate between models concerning the relative immunodominance of specific parasite products. PMID- 10831381 TI - Fasciola hepatica: an antigen fraction derived from newly excysted juveniles, containing an immunoreactive 32-kDa protein, induces strong protective immunity in rats. AB - Crude antigens of adult Fasciola hepatica and of newly excysted juveniles (NEJ) and a low-molecular-weight fraction of antigen from NEJs were tested for inducing protective immunity in rats. Two routes of vaccination were applied. The results showed that intraperitoneal vaccination induced significantly better protection (P <0.05) than intramuscular vaccination. Intraperitoneal vaccination with antigens from NEJs induced more effective protection: after challenge infection, rats that were so vaccinated had 92.6% (+/-2.5% SEM) fewer parasites in their liver and 57.3% (+/-13.3% SEM) fewer parasites penetrating the gut wall than control rats. Rats that were vaccinated with a low-molecular-weight fraction of antigen from NEJs were also highly protected against a challenge. F. hepatica antigens that are immunoreactive were identified on immunoblots, using sera collected from highly protected rats that had been vaccinated with NEJ antigens and also sera from cattle and rats that were experimentally infected with F. hepatica. The low-molecular-weight fraction of antigen from NEJs contained an immunodominant 32-kDa protein that was recognized by serum antibodies of vaccinated rats and immune cattle. This 32-kDa protein was not detected in partially purified antigens from adult flukes. We conclude that antigens of NEJs of F. hepatica, when injected intraperitoneally in rats, are highly protective. In particular, the 32-kDa protein contained in these antigens may be highly valuable for the development of an effective vaccine against F. hepatica. PMID- 10831382 TI - Onchocerca volvulus superoxide dismutase genes: identification of functional promoters for pre-mRNA transcripts which undergo trans-splicing. AB - The genes encoding three forms of superoxide dismutase, the cytosolic and extracellular CuZn superoxide dismutases and the mitochondrial Mn superoxide dismutase, were isolated from an Onchocerca volvulus lambda fix II genomic library. Genomic Southern blot analyses indicate single-copy genes in the O. volvulus genome. The O. volvulus cytosolic and extracellular CuZnSOD genes (Ov sod-1 and Ov-sod-2) are separated by 0.8 kb of sequence and are convergently transcribed. Since the transcripts from all three sod genes are trans-spliced, the transcription start point of each gene was determined in a heterologous system that lacks trans-splicing machinery by in vitro transcription using Drosophila embryo nuclear extracts, followed by primer extension experiments. The ability of the 5' flanking region of the genes encoding the three Ov-SODs to promote transcription was further examined in transient transfections of Chinese hamster ovary cells. In firefly luciferase reporter assays, the Ov-sod-1 and -2 and the MnSOD (Ov-sod-3) gene promoters showed minimal, strong, and moderate levels of activity in these cells, respectively. Both Ov-sod-2 and -3 gene promoter regions showed an initial increase in activity in response to 5' deletions. The results from the in vitro transcription experiments and the luciferase reporter assays were consistent and suggest the presence of Inr-like elements in the promoter regions of the Ov-sod genes. PMID- 10831383 TI - Paragonimus westermani: a cytosolic glutathione S-transferase of a sigma-class in adult stage. AB - We purified cytosolic glutathione S-transferase (GST) of adult Paragonimus westermani monitoring its activity with 1-chloro-2,4-dinitrobenzene (CDNB). The enzyme was purified 18.4-fold to electrophoretic homogeneity with 21% recovery rate through a three-step procedure. The purified enzyme (Pw28GST) has a subunit molecular weight of 28 kDa with an isoelectric point at 4.6. Monoclonal antibody (anti-Pw28GST) against Pw28GST did not cross-react with GSTs from other helminths. cDNA library was constructed in lambdaZAP II bacteriophage and screened with anti-Pw28GST. The corresponding gene containing a single open reading frame of 804 bp encoded 211 amino acids. The predicted amino acid sequence exhibited a higher homology with catalytic domain near N-terminus of class sigma GSTs (58%) than with schistosome 28-kDa GSTs (45-41%) or with class sigma GSTs themselves (33-31%). The sequence contained both Tyr-6 and Tyr-10 that are highly conserved in mammalian and helminth GSTs. The apparent K(m) value of a recombinant enzyme was 0.78 mM. Both native and recombinant enzymes showed the highest activity against CDNB, relatively weak activity against ethacrynic acid and reactive carbonyls, and no activity against epoxy-3-(p-nitrophenoxy)-propane. The activities were inhibited by bromosulfophthalein, cibacron blue, and albendazole, but not by praziquantel. These findings indicate that adult P. westermani has a class sigma GST. PMID- 10831384 TI - Plasmodium yoelii nigeriensis (MDR)-efficacy of oral pyronaridine against multidrug-resistant malaria in Swiss mice. AB - A multidrug-resistant strain of Plasmodium yoelii nigeriensis (MDR) showing a wide spectrum of resistance to chloroquine, amodiaquine, mepacrine, mefloquine, halofantrine, quinine, and quinidine was used in this study for in vivo evaluation of the blood schizontocidal activity of pyronaridine, a topoisomerase II inhibitor, in Swiss mice. The parasite produces 100% lethal infection in mice. The drug was administered orally once a day from day 0 onward. The initial studies showed that low doses of pyronaridine (0.625 to 5.0 mg base/kg x9 days) did not completely control blood-induced P. yoelii nigeriensis infection. Finally a series of doses of pyronaridine ranging from 1.25 to 30.0 mg/kg administered orally for 7 consecutive days were evaluated and in spite of high level of resistance to standard antimalarials, the parasite P. yoelii nigeriensis has shown complete susceptibility to pyronaridine (15 mg/kg dose x7 days). The present paper also compares the merits of a single MDR strain vs a battery of different resistant lines for quick antimalarials screening. PMID- 10831385 TI - Plasmodium falciparum: merozoite surface proteins 1 and 2 are not posttranslationally modified by classical N- or O-glycans. PMID- 10831386 TI - Aminothiol multidentate chelators against Chagas disease. AB - Three compounds of an aminothiol family of iron chelators were examined for activity against trypomastigote (human) and epimastigote (vector) forms of Trypanosoma cruzi: tetraethyl and tetramethyl derivatives of ethane-1,2-bis (N-1 amino-3-ethyl butyl-3-thiol) (BAT-TE and BAT-TM) and N',N',N'-tris-(2-methyl-2 mercaptopriopyl)- 1,4,7-triazacyclonane (TAT). BAT-TE at 270 microM completely arrested the growth of trypomastigote forms in mouse blood stored at 4 degrees C for 24 h (IC(50) 67.7+/-7 microM), while BAT-TM arrested growth at 630 microM (IC(50) 158+/-17 microM) and TAT at concentrations >800 microM (IC(50) 415+/-55 microM). In T. cruzi-infected mice, BAT-TE and BAT-TM had no anti-trypanosomal activity in doses up to 200 mg/kg, whether the route of administration was intraperitoneal or oral, and TAT was not tested due to insufficient quantity. TAT had an IC(50) of 52+/-7 microM against the epimastigote forms while BAT-TM and BAT-TE were inhibitory only at concentrations >250 microM. The trypanocidal activity of BAT derivatives in blood stored at 4 degrees C makes these compounds potential candidates for the purpose of clearing donated blood of trypomastigotes. PMID- 10831387 TI - The new subfamily of cathepsin-Z-like protease genes includes Tc-cpz-1, a cysteine protease gene expressed in Toxocara canis adults and infective stage larvae. PMID- 10831388 TI - Trypanosoma cruzi: roles for perforin-dependent and perforin-independent immune mechanisms in acute resistance. AB - CD8+ T cells have been shown to be required for acute resistance to infection with the protozoan parasite, Trypanosoma cruzi, the causative agent of Chagas' disease. However, to date, the mechanism by which CD8+ T cells mediate protection in vivo has not been determined. While CD8+ T cells can exhibit cytolytic function, they also secrete cytokines such as IFN-gamma, which is known to mediate protection against T. cruzi infections. To determine whether cytolysis is an important effector function in vivo, we have compared outcomes of T. cruzi infection in normal and perforin-deficient mice. Our results indicate that while perforin-dependent cytolytic mechanisms clearly make a major contribution to acute resistance to T. cruzi infection, this contribution may be strain and challenge dose-dependent, since perforin-deficient mice challenged with lower doses of a less virulent strain survived and were subsequently resistant to challenge with virulent organisms. In vivo depletion studies demonstrated that survival of perforin-deficient mice challenged with low doses of T. cruzi requires both CD4+ and CD8+ T cells and is dependent on IFN-gamma secretion. These studies document the participation of both perforin-dependent cytotoxic and perforin-independent, IFN-gamma-dependent immune mechanisms in acute resistance to T. cruzi infection. PMID- 10831389 TI - Schistosoma mansoni: effects of serotonin and serotonin receptor antagonists on motility and length of primary sporocysts in vitro. AB - The effects of serotonin (5-hydroxytryptamine; 5-HT) on in vitro transformed primary sporocysts of Schistosoma mansoni were investigated. Serotonin treatment significantly increased parasite motility (percentage of motile sporocysts) and length at concentrations as low as 1 microM. These effects were mimicked by the 5 HT agonist tryptamine, albeit with 10- to 100-fold less potency. The effects of 10 microM 5-HT on sporocyst motility were observed within 15 min posttreatment and on parasite length by 6 h posttreatment, and both effects were stable for up to 48 h. Receptor antagonists with varying affinities for defined vertebrate neurotransmitter receptor subtypes were examined for their effects on parasite behavior in the absence and presence of 10 microM 5-HT. In the absence of 5-HT, only methiothepin significantly inhibited normal parasite growth after 48 h of incubation. In the presence of 10 microM 5-HT, the serotonin receptor antagonists mianserin, ketanserin (both at 100 microM), and methiothepin (at 10 microM) significantly inhibited 5-HT-induced lengthening of primary sporocysts, while 3 tropanyl-indole-3-carboxylate and chlorpromazine had no significant effect. The effects of these same drugs on parasite motility were also examined. In the absence of 5-HT, 10 microM chlorpromazine increased parasite motility, while the other antagonists had no effect. When sporocysts were treated with 10 microM 5-HT for 2 h in the continued presence of antagonist, 100 microM mianserin, ketanserin, 3-tropanyl-indole-3-carboxylate, and 10 microM methiothepin inhibited 5-HT induced increases in parasite motility, while 10 microM chlorpromazine had no effect. These results show that primary sporocysts of S. mansoni exhibit behavioral responses to serotonin much like adult stages of this parasite. Furthermore, these responses appear to be mediated via receptors with pharmacological similarities to those previously described in adult worms. PMID- 10831390 TI - Fasciola hepatica: influence of gender and liver biotransformations on flukicide treatment efficacy of rats infested and cured with either clorsulon/ivermectin or triclabendazole. AB - Two fasciolicide preparations have been compared in 130 rats experimentally infected with Fasciola hepatica. Parasitological, immunological, and biochemical parameters have been followed to monitor the efficacy of the treatments. While Fascinex (triclabendazole) efficiently cured both male and female rats when administered as soon as 4 weeks postinfection, treatment with Ivomec-D (clorsulon + ivermectin) displayed a low efficacy on either male or female rats at this time point (54 and 0%, respectively). Moreover, when administered 8 weeks postinfection, the Ivomec-D treatment proved highly efficient on male rats while it displayed little effect on the female population (100 and 53%, respectively). This unexpected result has been related to an overexpression of a P4503A isoform that is observed only in females that have been treated with Ivomec-D. The influence of this P4503A cytochrome on drug metabolism and the need for the incorporation of both genders in clinical trials are discussed. PMID- 10831391 TI - Pathogenic leishmania secrete antigenically related chitinases which are encoded by a highly conserved gene locus. AB - Recently, we identified and characterized a single-copy chitinase gene (LdCht1) from Leishmania donovani, a protozoan pathogen of humans. It has been hypothesized that this parasite enzyme plays a critical role in the survival of all Leishmania species within their sandfly vectors and for their transmission to humans. Thus, in the current study, pulse-field gel electrophoresis and Southern hybridization with the LdCht1 gene probe were used to demonstrate that this chitinase gene has been conserved across species lines of various pathogenic Leishmania. Further, immunoprecipitation and enzyme activity assays using an anti LdCht1-peptide serum were used to show that the chitinases produced and released by this group of parasites possessed both highly conserved antigenic epitopes and enzyme activities. Results of these studies demonstrate that the chitinase gene locus and enzyme activity have been conserved across species lines among this group of human pathogens. PMID- 10831392 TI - The intraperitoneal Plasmodium berghei-Pasteur infection of Swiss mice is not a system that is able to detect the antiplasmodial activity in the Pothomorphe plant extracts that are used as antimalarials in Brazilian endemic areas. AB - The antimalarial activity of the hexane and methanol extracts derived from the Brazilian plants Pothomorphe peltata and Pothomorphe umbellata-whose leaves are popularly employed in medicinal folk remedies for the treatment of malaria-was assessed through in vivo tests with the Peters method. The extracts were delivered to Plasmodium berghei-infected mice via the oral or the subcutaneous route. A suppressive effect on the parasitemia seemed to be evident when data regarding the intraperitoneal injection of Pothomorphe umbellata extracts were analyzed. However, a definitive conclusion on an effective antimalarial activity is not possible, as two distinct-"standard" and "slow"-patterns of parasitemia occurring at similar frequencies in both treated and untreated intraperitoneally infected mice were observed. Nevertheless, the existence of two distinct profiles of parasitemia was not clear among the animals that were infected via the intravenous route. These data indicate the need for further studies on the biological features of the host/parasite interaction in the intraperitoneally P. berghei-infected Swiss mice system to standardize the model and to improve its usefulness in the screening of antimalarial compounds. PMID- 10831393 TI - Cloning and characterization of Leishmania donovani telomeres. AB - We describe here the cloning and sequence characterization of the absolute termini of several telomeres from the human parasite Leishmania donovani using a vector-adapter protocol. The 3' protruding strand of L. donovani telomeres terminates with the sequence 5'-GGTTAGGGT-OH 3'. This single-stranded sequence is adjacent to tandemly repeated blocks of double-stranded sequence consisting of variable numbers of the hexameric repeat 5'-TAGGGT-3', variable numbers of an octameric repeat 5'-TGGTCATG-3', and a single 62-bp sequence, in that order. A number of additional, more chromosome-internal, nonrepeated sequences were found adjacent to the telomere sequences. Hybridization analyses indicated that some of these telomere adjacent sequences are found on all L. donovani chromosomes, some are more abundant on certain subsets of chromosomes, and some are unique to individual chromosomes. PMID- 10831394 TI - Plasmodium berghei: the antimalarial activity of albendazole in rats is mediated via effects on the hematopoietic system. PMID- 10831395 TI - Plasmodium falciparum: polymorphism in regions II and III of the knob-associated histidine-rich protein gene from two areas of different endemicity. PMID- 10831396 TI - Plasmodium sp.: optimal protocols for PCR detection of low parasite numbers from mosquito (Anopheles sp.) samples. PMID- 10831397 TI - Genome display and typing of Plasmodium parasites using anchored PolyA and PolyT oligonucleotides. PMID- 10831398 TI - Testing linkage disequilibrium in sibships. AB - We describe the use of multivariate regression for testing allelic association in the presence of linkage, using marker genotype data from sibships. The test is valid, provided that the correct mean structure is modeled but does not require the correlation structure within families to be specified. The test can be implemented using standard statistical software such as the SAS programming language. In a simulation study, we evaluated this new test in comparison with one from a standard, matched-case-control analysis. First, we noted that the genetic effect needed to be quite extreme before residual familial correlation due to linkage led to false inference using the standard, matched-pair analysis. Second, we showed that under examples of extreme residual familial correlation, the new test had the correct test size. Third, we found that the test was more powerful than the sibship disequilibrium test of Horvath and Laird. Finally, we concluded that although the standard analysis may lead to correct inference for practical purposes, the new test is valid, even under extreme residual familial correlation and with no cost in power at the causal locus. PMID- 10831400 TI - A comparison of methods for counting viruses in aquatic systems. AB - In this study, we compared different methods-including transmission electron microscopy-and various nucleic acid labeling methods in which we used the fluorochromes 4',6'-diamidino-2-phenylindole (DAPI), 4-[3-methyl-2,3-dihydro (benzo-1, 3-oxazole)-2-methylmethyledene]-1-(3'-trimethyl ammoniumpropyl) quinilinium diioide (YOPRO-1), and SYBR Green I, which can be detected by epifluorescence microscopy (EM), for counting viruses in samples obtained from freshwater ecosystems whose trophic status varied and from a culture of T7 phages. From a quantitative and qualitative viewpoint, our results showed that the greatest efficiency for all ecosystems was obtained when we used the EM counting protocol in which YOPRO-1 was the label, as this fluorochrome exhibited strong and very stable fluorescence. A modification of the original protocol in which YOPRO-1 was used is recommended, because this modification makes the protocol faster and allows it to be used for routine analysis of fixed samples. Because SYBR Green I fades very quickly, the use of this fluorochrome is not recommended for systems in which the viral content is very high (>10(8) particles/ml), such as treated domestic sewage effluents. Experiments in which we used DNase and RNase revealed that the number of viruses determined by EM was slightly overestimated (by approximately 15%) because of interference caused by the presence of free nucleic acids. PMID- 10831399 TI - N-myc downstream-regulated gene 1 is mutated in hereditary motor and sensory neuropathy-Lom. AB - Hereditary motor and sensory neuropathies, to which Charcot-Marie-Tooth (CMT) disease belongs, are a common cause of disability in adulthood. Growing awareness that axonal loss, rather than demyelination per se, is responsible for the neurological deficit in demyelinating CMT disease has focused research on the mechanisms of early development, cell differentiation, and cell-cell interactions in the peripheral nervous system. Autosomal recessive peripheral neuropathies are relatively rare but are clinically more severe than autosomal dominant forms of CMT, and understanding their molecular basis may provide a new perspective on these mechanisms. Here we report the identification of the gene responsible for hereditary motor and sensory neuropathy-Lom (HMSNL). HMSNL shows features of Schwann-cell dysfunction and a concomitant early axonal involvement, suggesting that impaired axon-glia interactions play a major role in its pathogenesis. The gene was previously mapped to 8q24.3, where conserved disease haplotypes suggested genetic homogeneity and a single founder mutation. We have reduced the HMSNL interval to 200 kb and have characterized it by means of large-scale genomic sequencing. Sequence analysis of two genes located in the critical region identified the founder HMSNL mutation: a premature-termination codon at position 148 of the N-myc downstream-regulated gene 1 (NDRG1). NDRG1 is ubiquitously expressed and has been proposed to play a role in growth arrest and cell differentiation, possibly as a signaling protein shuttling between the cytoplasm and the nucleus. We have studied expression in peripheral nerve and have detected particularly high levels in the Schwann cell. Taken together, these findings point to NDRG1 having a role in the peripheral nervous system, possibly in the Schwann-cell signaling necessary for axonal survival. PMID- 10831401 TI - Distribution of aldoxime dehydratase in microorganisms. AB - The distribution of phenylacetaldoxime-degrading and pyridine-3-aldoxime degrading ability was examined with intact cells of 975 microorganisms, including 45 genera of bacteria, 11 genera of actinomyces, 22 genera of yeasts, and 37 genera of fungi, by monitoring the decrease of the aldoximes by high-pressure liquid chromatography. The abilities were found to be widely distributed in bacteria, actinomyces, fungi, and some yeasts: 98 and 107 strains degraded phenylacetaldoxime and pyridine-3-aldoxime, respectively. All of the active strains exhibited not only the aldoxime-dehydration activity to form nitrile but also nitrile-hydrolyzing activity. On the other hand, all of 19 nitrile-degrading microorganisms (13 species, 7 genera) were found to exhibit aldoxime dehydration activity. It is shown that aldoxime dehydratase and nitrile-hydrolyzing activities are widely distributed among 188 aldoxime and 19 nitrile degraders and that the enzymes were induced by aldoximes or nitriles. PMID- 10831402 TI - Reductive dehalogenation of trichloroacetic acid by Trichlorobacter thiogenes gen. nov., sp. nov. AB - A bacterium able to grow via reductive dechlorination of trichloroacetate was isolated from anaerobic soil enrichments. The isolate, designated strain K1, is a member of the delta proteobacteria and is related to other known sulfur and ferric iron reducers. In anaerobic mineral media supplemented with acetate and trichloroacetate, its doubling time was 6 h. Alternative electron donor and acceptors were acetoin and sulfur or fumarate, respectively. Trichloroacetate dehalogenation activity was constitutively present, and the dechlorination product was dichloroacetate and chloride. Trichloroacetate conversion seemed to be coupled to a novel sulfur-sulfide redox cycle, which shuttled electrons from acetate oxidation to trichloroacetate reduction. In view of its unique physiological characteristics, the name Trichlorobacter thiogenes is suggested for strain K1. PMID- 10831403 TI - Production of exopolysaccharide by Lactobacillus rhamnosus R and analysis of its enzymatic degradation during prolonged fermentation. AB - The potential of Lactobacillus rhamnosus R for producing exopolysaccharide (EPS) when grown on basal minimum medium supplemented with glucose or lactose was investigated. EPS production by L. rhamnosus R is partially growth associated and about 500 mg of EPS per liter was synthesized with both sugars. The product yield coefficient (Y(EPS/S)) was 3.15 (0.0315 g of EPS [g of lactose](-1)) and 2.88 (0.0288 g of EPS [g of glucose](-1)). It was clearly shown that the amount of EPS produced declined upon prolonged fermentation. Degradation of EPS in fermentation processes was also assessed by measuring its molecular weights and viscosities. As these reductions might have a negative effect on the yield and viscosifying properties of EPS, it was essential to examine possible causes related to this breakdown. The decrease in viscosities and molecular weights of EPS withdrawn at different cultivation times permitted us to suspect the presence of a depolymerizing enzyme in the fermentation medium. Our study on enzymatic production profiles showed a large spectrum of glycohydrolases (alpha-D glucosidase, beta-D-glucosidase, alpha-D-galactosidase, beta-D-galactosidase, beta-D-glucuronidase, and some traces of alpha-L-rhamnosidase). These enzymes were localized, two of them (alpha-D-glucosidase and beta-D-glucuronidase) were partially purified and characterized. When incubated with EPS, these enzymes were capable of lowering the viscosity of the polymer as well as liberating some reducing sugars. Upon prolonged incubation (27 h), the loss of viscosity was increased up to 33%. PMID- 10831404 TI - Bioconversion of ferulic acid into vanillic acid by means of a vanillate-negative mutant of Pseudomonas fluorescens strain BF13. AB - From a ferulic-acid-degrading Pseudomonas fluorescens strain (BF13), we have isolated a transposon mutant, which retained the ability to bioconvert ferulic acid into vanillic acid but lost the ability to further degrade the latter acid. The mutant, BF13-97, was very stable, and therefore it was suitable to be used as a biocatalyst for the preparative synthesis of vanillic acid from ferulic acid. By use of resting cells we determined the effect on the bioconversion rate of several parameters, such as the addition of nutritional factors, the concentration of the biomass, and the carbon source on which the biomass was grown. The optimal yield of vanillic acid was obtained with cells pregrown on M9 medium containing p-coumaric acid (0.1% [wt/vol]) as a sole carbon source and yeast extract (0.001% [wt/vol]) as a source of nutritional factors. Under these conditions, 1 mg (wet weight) of biomass produced 0.23 mg of vanillic acid per h. The genomic region of BF13-97 flanking the transposon's site of insertion was cloned and sequenced revealing two open reading frames of 1,062 (vanA) and 954 (vanB) bp, respectively. The van genes are organized in a cluster and encode the subunits of the vanillate-O-demethylase, which catalyzes the first step of the vanillate catabolism. Amino acid sequences deduced from vanA and vanB genes were shown to have high identity with known VanAs and VanBs from Pseudomonas and Acinetobacter spp. Highly conserved regions known to exist in class IA oxygenases were also found in the vanillate-O-demethylase components from P. fluorescens BF13. The terminal oxygenase VanA is characterized by a conserved Rieske-type [2Fe-2S](R) ligand center. The reductase VanB contains a plant-type ferredoxin [2Fe-2S](Fd), flavin mononucleotide, and NAD-ribose binding domains which are located in its C-terminal and N-terminal halves, respectively. Transfer of wild type vanAB genes to BF13-97 complemented this mutant, which recovered its ability to grow on either vanillic or ferulic acid. PMID- 10831405 TI - Molecular cloning, sequencing, and expression of omp-40, the gene coding for the major outer membrane protein from the acidophilic bacterium Thiobacillus ferrooxidans. AB - Thiobacillus ferrooxidans is one of the chemolithoautotrophic bacteria important in industrial biomining operations. Some of the surface components of this microorganism are probably involved in adaptation to their acidic environment and in bacterium-mineral interactions. We have isolated and characterized omp40, the gene coding for the major outer membrane protein from T. ferrooxidans. The deduced amino acid sequence of the Omp40 protein has 382 amino acids and a calculated molecular weight of 40,095.7. Omp40 forms an oligomeric structure of about 120 kDa that dissociates into the monomer (40 kDa) by heating in the presence of sodium dodecyl sulfate. The degree of identity of Omp40 amino acid sequence to porins from enterobacteria was only 22%. Nevertheless, multiple alignments of this sequence with those from several OmpC porins showed several important features conserved in the T. ferrooxidans surface protein, such as the approximate locations of 16 transmembrane beta strands, eight loops, including a large external L3 loop, and eight turns which allowed us to propose a putative 16 stranded beta-barrel porin structure for the protein. These results together with the previously known capacity of Omp40 to form ion channels in planar lipid bilayers strongly support its role as a porin in this chemolithoautotrophic acidophilic microorganism. Some characteristics of the Omp40 protein, such as the presence of a putative L3 loop with an estimated isoelectric point of 7.21 allow us to speculate that this can be the result of an adaptation of the acidophilic T. ferrooxidans to prevent free movement of protons across its outer membrane. PMID- 10831406 TI - Gene cloning, sequencing, and inactivation of the branched-chain aminotransferase of Lactococcus lactis LM0230. AB - A branched-chain aminotransferase gene (ilvE) from Lactococcus lactis LM0230 was identified on a 9-kb chromosomal insert by complementation in Escherichia coli DL39. Sequencing of a 2.0-kbp fragment resulted in the identification of a 1,023 bp open reading frame that could encode a 340-amino-acid protein. Sequence analysis of the deduced amino acid sequence revealed 62% identity to IlvE of Haemophilus influenzae and high similarity to IlvEs from a variety of organisms found in GenBank classified as class IV aminotransferases. Under logarithmic growth in complex medium, ilvE is transcribed monocistronically as a 1.1-kb transcript. Hydrophobicity plot analysis of the deduced amino acid sequence and the lack of a signal peptide sequence suggest IlvE is a cytosolic protein. A derivative of LM0230 lacking IlvE activity was constructed by gene replacement. Comparison of the IlvE-deficient strain's ability to grow in defined media lacking an amino acid but containing its alpha-keto acid biosynthetic precursor to that of the wild-type strain indicated that IlvE is the only enzyme capable of synthesis of Ile and Val from their biosynthetic precursors. Comparison of the aminotransferase activity of the IlvE mutant to LM0230 revealed that the mutant retained <2, 4.5, 43, 40, and 76% of its aminotransferase activity with Ile, Val, Leu, Met, and Phe, respectively. No difference in growth or acidification rate between LM0230 and the IlvE-deficient strain was observed in milk. PMID- 10831407 TI - Dynamic changes of intracellular pH in individual lactic acid bacterium cells in response to a rapid drop in extracellular pH. AB - We describe the dynamics of changes in the intracellular pH (pH(i)) values of a number of lactic acid bacteria in response to a rapid drop in the extracellular pH (pH(ex)). Strains of Lactobacillus delbrueckii subsp. bulgaricus, Streptococcus thermophilus, and Lactococcus lactis were investigated. Listeria innocua, a gram-positive, non-lactic acid bacterium, was included for comparison. The method which we used was based on fluorescence ratio imaging of single cells, and it was therefore possible to describe variations in pH(i) within a population. The bacteria were immobilized on a membrane filter, placed in a closed perfusion chamber, and analyzed during a rapid decrease in the pH(ex) from 7.0 to 5.0. Under these conditions, the pH(i) of L. innocua remained neutral (between 7 and 8). In contrast, the pH(i) values of all of the strains of lactic acid bacteria investigated decreased to approximately 5.5 as the pH(ex) was decreased. No pronounced differences were observed between cells of the same strain harvested from the exponential and stationary phases. Small differences between species were observed with regard to the initial pH(i) at pH(ex) 7.0, while different kinetics of pH(i) regulation were observed in different species and also in different strains of S. thermophilus. PMID- 10831408 TI - Biotransformation of hydroxylaminobenzene and aminophenol by Pseudomonas putida 2NP8 cells grown in the presence of 3-nitrophenol. AB - Biotransformation products of hydroxylaminobenzene and aminophenol produced by 3 nitrophenol-grown cells of Pseudomonas putida 2NP8, a strain grown on 2- and 3 nitrophenol, were characterized. Ammonia, 2-aminophenol, 4-aminophenol, 4 benzoquinone, N-acetyl-4-aminophenol, N-acetyl-2-aminophenol, 2-aminophenoxazine 3-one, 4-hydroquinone, and catechol were produced from hydroxylaminobenzene. Ammonia, N-acetyl-2-aminophenol, and 2-aminophenoxazine-3-one were produced from 2-aminophenol. All of these metabolites were also found in the nitrobenzene transformation medium, and this demonstrated that they were metabolites of nitrobenzene transformation via hydroxylaminobenzene. Production of 2 aminophenoxazine-3-one indicated that oxidation of 2-aminophenol via imine occurred. Rapid release of ammonia from 2-aminophenol transformation indicated that hydrolysis of the imine intermediate was the dominant reaction. The low level of 2-aminophenoxazine-3-one indicated that formation of this compound was probably due to a spontaneous reaction accompanying oxidation of 2-aminophenol via imine. 4-Hydroquinone and catechol were reduction products of 2- and 4 benzoquinones. Based on these transformation products, we propose a new ammonia release pathway via oxidation of aminophenol to benzoquinone monoimine and subsequent hydrolysis for transformation of nitroaromatic compounds by 3 nitrophenol-grown cells of P. putida 2NP8. We propose a parallel mechanism for 3 nitrophenol degradation in P. putida 2NP8, in which all of the possible intermediates are postulated. PMID- 10831409 TI - Isolation and expression of lactate dehydrogenase genes from Rhizopus oryzae. AB - Rhizopus oryzae is used for industrial production of lactic acid, yet little is known about the genetics of this fungus. In this study I cloned two genes, ldhA and ldhB, which code for NAD(+)-dependent L-lactate dehydrogenases (LDH) (EC 1.1.1.27), from a lactic acid-producing strain of R. oryzae. These genes are similar to each other and exhibit more than 90% nucleotide sequence identity and they contain no introns. This is the first description of ldh genes in a fungus, and sequence comparisons revealed that these genes are distinct from previously isolated prokaryotic and eukaryotic ldh genes. Protein sequencing of the LDH isolated from R. oryzae during lactic acid production confirmed that ldhA codes for a 36-kDa protein that converts pyruvate to lactate. Production of LdhA was greatest when glucose was the carbon source, followed by xylose and trehalose; all of these sugars could be fermented to lactic acid. Transcripts from ldhB were not detected when R. oryzae was grown on any of these sugars but were present when R. oryzae was grown on glycerol, ethanol, and lactate. I hypothesize that ldhB encodes a second NAD(+)-dependent LDH that is capable of converting L lactate to pyruvate and is produced by cultures grown on these nonfermentable substrates. Both ldhA and ldhB restored fermentative growth to Escherichia coli (ldhA pfl) mutants so that they grew anaerobically and produced lactic acid. PMID- 10831410 TI - Molecular and physiological responses of two classes of marine chromophytic phytoplankton (Diatoms and prymnesiophytes) during the development of nutrient stimulated blooms. AB - Generic taxon-specific DNA probes that target an internal region of the gene (rbcL) encoding the large subunit of ribulose-1, 5-bisphosphate carboxylase/oxygenase (RubisCO) were developed for two groups of marine phytoplankton (diatoms and prymnesiophytes). The specificity and utility of the probes were evaluated in the laboratory and also during a 1-month mesocosm experiment in which we investigated the temporal variability in RubisCO gene expression and primary production in response to inorganic nutrient enrichment. We found that the onset of successive bloom events dominated by each of the two classes of chromophyte algae was associated with marked taxon-specific increases in rbcL transcription rates. These observations suggest that measurements of RubisCO gene expression can provide an early indicator of the development of phytoplankton blooms and may also be useful in predicting which taxa are likely to dominate a bloom. PMID- 10831411 TI - Osmoprotection by pipecolic acid in Sinorhizobium meliloti: specific effects of D and L isomers. AB - DL-Pipecolic acid (DL-PIP) promotes growth restoration of Sinorhizobium meliloti cells facing inhibitory hyperosmolarity. Surprisingly, D and L isomers of this imino acid supplied separately were not effective. The uptake of L-PIP was significantly favored in the presence of the D isomer and by a hyperosmotic stress. Chromatographic analysis of the intracellular solutes showed that stressed cells did not accumulate radiolabeled L-PIP. Rather, it participates in the synthesis of the main endogenous osmolytes (glutamate and the dipeptide N acetylglutaminylglutamine amide) during the lag phase, thus providing a means for the stressed cells to recover the osmotic balance. (13)C nuclear magnetic resonance analysis was used to determine the fate of D-PIP taken into the cells. In the absence of L-PIP, the imported D isomer was readily degraded. Supplied together with its L isomer, D-PIP was accumulated temporarily and thus might contribute together with the endogenous osmolytes to enhance the internal osmotic strength. Furthermore, it started to disappear from the cytosol when the L isomer was no longer available in the culture medium (during the late exponential phase of growth). Together, these results show an uncommon mechanism of protection of osmotically stressed cells of S. meliloti. It was proved, for the first time, that the presence of the two isomers of the same molecule is necessary for it to manifest an osmoprotective activity. Indeed, D-PIP seems to play a major role in cellular osmoadaptation through both its own accumulation and improvement of the utilization of the L isomer as an immediate precursor of endogenous osmolytes. PMID- 10831412 TI - Prey range characterization, ribotyping, and diversity of soil and rhizosphere Bdellovibrio spp. isolated on phytopathogenic bacteria. AB - Thirty new Bdellovibrio strains were isolated from an agricultural soil and from the rhizosphere of plants grown in that soil. Using a combined molecular and culture-based approach, we found that the soil bdellovibrios included subpopulations of organisms that differed from rhizosphere bdellovibrios. Thirteen soil and seven common bean rhizosphere Bdellovibrio strains were isolated when Pseudomonas corrugata was used as prey; seven and two soil strains were isolated when Erwinia carotovora subsp. carotovora and Agrobacterium tumefaciens, respectively, were used as prey; and one tomato rhizosphere strain was isolated when A. tumefaciens was used as prey. In soil and in the rhizosphere, depending on the prey cells used, the concentrations of bdellovibrios were between 3 x 10(2) to 6 x 10(3) and 2.8 x 10(2) to 2.3 x 10(4) PFU g(-1). A prey range analysis of five soil and rhizosphere Bdellovibrio isolates performed with 22 substrate species, most of which were plant-pathogenic and plant growth-enhancing bacteria, revealed unique utilization patterns and differences between closely related prey cells. An approximately 830-bp fragment of the 16S rRNA genes of all of the Bdellovibrio strains used was obtained by PCR amplification by using a Bdellovibrio-specific primer combination. Soil and common bean rhizosphere strains produced two and one restriction patterns for this PCR product, respectively. The 16S rRNA genes of three soil isolates and three root-associated isolates were sequenced. One soil isolate belonged to the Bdellovibrio stolpii-Bdellovibrio starrii clade, while all of the other isolates clustered with Bdellovibrio bacteriovorus and formed two distantly related, heterogeneous groups. PMID- 10831413 TI - Anaerobic-aerobic process for microbial degradation of tetrabromobisphenol A. AB - Tetrabromobisphenol A (TBBPA) is a flame retardant that is used as an additive during manufacturing of plastic polymers and electronic circuit boards. Little is known about the fate of this compound in the environment. In the current study we investigated biodegradation of TBBPA, as well as 2,4,6-tribromophenol (TBP), in slurry of anaerobic sediment from a wet ephemeral desert stream bed contaminated with chemical industry waste. Anaerobic incubation of the sediment with TBBPA and peptone-tryptone-glucose-yeast extract medium resulted in a 80% decrease in the TBBPA concentration and accumulation of a single metabolite. This metabolite was identified by gas chromatography-mass spectrometry (GC-MS) as nonbrominated bisphenol A (BPA). On the other hand, TBP was reductively dehalogenated to phenol, which was further metabolized under anaerobic conditions. BPA persisted in the anaerobic slurry but was degraded aerobically. A gram-negative bacterium (strain WH1) was isolated from the contaminated soil, and under aerobic conditions this organism could use BPA as a sole carbon and energy source. During degradation of BPA two metabolites were detected in the culture medium, and these metabolites were identified by GC-MS and high-performance liquid chromatography as 4-hydroxybenzoic acid and 4-hydroxyacetophenone. Both of those compounds were utilized by WH1 as carbon and energy sources. Our findings demonstrate that it may be possible to use a sequential anaerobic-aerobic process to completely degrade TBBPA in contaminated soils. PMID- 10831414 TI - The macrocyclic peptide antibiotic micrococcin P(1) is secreted by the food-borne bacterium Staphylococcus equorum WS 2733 and inhibits Listeria monocytogenes on soft cheese. AB - Staphylococcus equorum WS 2733 was found to produce a substance exhibiting a bacteriostatic effect on a variety of gram-positive bacteria. The metabolite was purified to homogeneity by ammonium sulfate precipitation and semipreparative reversed-phase high-performance liquid chromatography. Electrospray mass spectrometry confirmed the high purity of the compound and revealed a molecular mass of 1,143 Da. By two-dimensional nuclear magnetic resonance spectroscopy the substance was identified as micrococcin P(1) which is a macrocyclic peptide antibiotic that has not yet been reported for the genus Staphylococcus. A total of 95 out of 95 Listeria strains and 130 out of 135 other gram-positive bacteria were inhibited by this substance, while none of 37 gram-negative bacteria were affected. The antilisterial potential of this food-grade strain as a protective starter culture was evaluated by its in situ application in cheese-ripening experiments under laboratory conditions. A remarkable growth reduction of Listeria monocytogenes could be achieved compared to control cheese ripened with a nonbacteriocinogenic type strain of Staphylococcus equorum. In order to prove that inhibition was due to micrococcin P(1), a micrococcin-deficient mutant was constructed which did not inhibit L. monocytogenes in cheese-ripening experiments. PMID- 10831415 TI - Phylogenetic relationships of Cryptosporidium parasites based on the 70 kilodalton heat shock protein (HSP70) gene. AB - We have characterized the nucleotide sequences of the 70-kDa heat shock protein (HSP70) genes of Cryptosporidium baileyi, C. felis, C. meleagridis, C. muris, C. serpentis, C. wrairi, and C. parvum from various animals. Results of the phylogenetic analysis revealed the presence of several genetically distinct species in the genus Cryptosporidium and eight distinct genotypes within the species C. parvum. Some of the latter may represent cryptic species. The phylogenetic tree constructed from these sequences is in agreement with our previous results based on the small-subunit rRNA genes of Cryptosporidium parasites. The Cryptosporidium species formed two major clades: isolates of C. muris and C. serpentis formed the first major group, while isolates of C. felis, C. meleagridis, C. wrairi, and eight genotypes of C. parvum formed the second major group. Sequence variations were also observed between C. muris isolates from ruminants and rodents. The HSP70 gene provides another useful locus for phylogenetic analysis of the genus Cryptosporidium. PMID- 10831416 TI - A Mycobacterium strain with extended capacities for degradation of gasoline hydrocarbons. AB - A bacterial strain (strain IFP 2173) was selected from a gasoline-polluted aquifer on the basis of its capacity to use 2,2, 4-trimethylpentane (isooctane) as a sole carbon and energy source. This isolate, the first isolate with this capacity to be characterized, was identified by 16S ribosomal DNA analysis, and 100% sequence identity with a reference strain of Mycobacterium austroafricanum was found. Mycobacterium sp. strain IFP 2173 used an unusually wide spectrum of hydrocarbons as growth substrates, including n-alkanes and multimethyl substituted isoalkanes with chains ranging from 5 to 16 carbon atoms long, as well as substituted monoaromatic hydrocarbons. It also attacked ethers, such as methyl t-butyl ether. During growth on gasoline, it degraded 86% of the substrate. Our results indicated that strain IFP 2173 was capable of degrading 3 methyl groups, possibly by a carboxylation and deacetylation mechanism. Evidence that it attacked the quaternary carbon atom structure by an as-yet-undefined mechanism during growth on 2,2,4-trimethylpentane and 2,2-dimethylpentane was also obtained. PMID- 10831417 TI - Bacterial community structure and physiological state within an industrial phenol bioremediation system. AB - The structure of bacterial populations in specific compartments of an operational industrial phenol remediation system was assessed to examine bacterial community diversity, distribution, and physiological state with respect to the remediation of phenolic polluted wastewater. Rapid community fingerprinting by PCR-based denaturing gradient gel electrophoresis (DGGE) of 16S rDNA indicated highly structured bacterial communities residing in all nine compartments of the treatment plant and not exclusively within the Vitox biological reactor. Whole cell targeting by fluorescent in situ hybridization with specific oligonucleotides (directed to the alpha, beta and gamma subclasses of the class Proteobacteria [alpha-, beta-, and gamma-Proteobacteria, respectively], the Cytophaga-Flavobacterium group, and the Pseudomonas group) tended to mirror gross changes in bacterial community composition when compared with DGGE community fingerprinting. At the whole-cell level, the treatment compartments were numerically dominated by cells assigned to the Cytophaga-Flavobacterium group and to the gamma-Proteobacteria. The alpha subclass Proteobacteria were of low relative abundance throughout the treatment system whilst the beta subclass of the Proteobacteria exhibited local dominance in several of the processing compartments. Quantitative image analyses of cellular fluorescence was used as an indicator of physiological state within the populations probed with rDNA. For cells hybridized with EUB338, the mean fluorescence per cell decreased with increasing phenolic concentration, indicating the strong influence of the primary pollutant upon cellular rRNA content. The gamma subclass of the Proteobacteria had a ribosome content which correlated positively with total phenolics and thiocyanate. While members of the Cytophaga-Flavobacterium group were numerically dominant in the processing system, their abundance and ribosome content data for individual populations did not correlate with any of the measured chemical parameters. The potential importance of the gamma-Proteobacteria and the Cytophaga-Flavobacteria during this bioremediation process was highlighted. PMID- 10831418 TI - Isolation and characterization of Desulfovibrio dechloracetivorans sp. nov., a marine dechlorinating bacterium growing by coupling the oxidation of acetate to the reductive dechlorination of 2-chlorophenol. AB - Strain SF3, a gram-negative, anaerobic, motile, short curved rod that grows by coupling the reductive dechlorination of 2-chlorophenol (2-CP) to the oxidation of acetate, was isolated from San Francisco Bay sediment. Strain SF3 grew at concentrations of NaCl ranging from 0.16 to 2.5%, but concentrations of KCl above 0. 32% inhibited growth. The isolate used acetate, fumarate, lactate, propionate, pyruvate, alanine, and ethanol as electron donors for growth coupled to reductive dechlorination. Among the halogenated aromatic compounds tested, only the ortho position of chlorophenols was reductively dechlorinated, and additional chlorines at other positions blocked ortho dechlorination. Sulfate, sulfite, thiosulfate, and nitrate were also used as electron acceptors for growth. The optimal temperature for growth was 30 degrees C, and no growth or dechlorination activity was observed at 37 degrees C. Growth by reductive dechlorination was revealed by a growth yield of about 1 g of protein per mol of 2-CP dechlorinated, and about 2.7 g of protein per mole of 2,6-dichlorophenol dechlorinated. The physiological features and 16S ribosomal DNA sequence suggest that the organism is a novel species of the genus Desulfovibrio and which we have designated Desulfovibrio dechloracetivorans. The unusual physiological feature of this strain is that it uses acetate as an electron donor and carbon source for growth with 2-CP but not with sulfate. PMID- 10831419 TI - Differential effects of permeating and nonpermeating solutes on the fatty acid composition of Pseudomonas putida. AB - We examined the effect of reduced water availability on the fatty acid composition of Pseudomonas putida strain mt-2 grown in a defined medium in which the water potential was lowered with the permeating solutes NaCl or polyethylene glycol (PEG) with a molecular weight of 200 (PEG 200) or the nonpermeating solute PEG 8000. Transmission electron microscopy showed that -1.0-MPa PEG 8000-treated cells had convoluted outer membranes, whereas -1.0-MPa NaCl-treated or control cells did not. At the range of water potential (-0.25 to -1.5 MPa) that we examined, reduced water availability imposed by PEG 8000, but not by NaCl or PEG 200, significantly altered the amounts of trans and cis isomers of monounsaturated fatty acids that were present in whole-cell fatty acid extracts. Cells grown in basal medium or under the -0.25-MPa water potential imposed by NaCl or PEG 200 had a higher trans:cis ratio than -0.25-MPa PEG 8000-treated cells. As the water potential was lowered further with PEG 8000 amendments, there was an increase in the amount of trans isomers, resulting in a higher trans:cis ratio. Similar results were observed in cells grown physically separated from PEG 8000, indicating that these changes were not due to PEG toxicity. When cells grown in -1.5-MPa PEG 8000 amendments were exposed to a rapid water potential increase of 1.5 MPa or to a thermodynamically equivalent concentration of the permeating solute, NaCl, there was a decrease in the amount of trans fatty acids with a corresponding increase in the cis isomer. The decrease in the trans/cis ratio following hypoosomotic shock did not occur in the presence of the lipid synthesis inhibitor cerulenin or the growth inhibitors chloramphenicol and rifampicin, which indicates a constitutively operating enzyme system. These results indicate that thermodynamically equivalent concentrations of permeating and nonpermeating solutes have unique effects on membrane fatty acid composition. PMID- 10831420 TI - Effect of temperature and salinity stress on growth and lipid composition of Shewanella gelidimarina. AB - The maximum growth temperature, the optimal growth temperature, and the estimated normal physiological range for growth of Shewanella gelidimarina are functions of water activity (a(w)), which can be manipulated by changing the concentration of sodium chloride. The growth temperatures at the boundaries of the normal physiological range for growth were characterized by increased variability in fatty acid composition. Under hyper- and hypoosmotic stress conditions at an a(w) of 0.993 (1.0% [wt/vol] NaCl) and at an a(w) of 0.977 (4.0% [wt/vol] NaCl) the proportion of certain fatty acids (monounsaturated and branched-chain fatty acids) was highly regulated and was inversely related to the growth rate over the entire temperature range. The physical states of lipids extracted from samples grown at stressful a(w) values at the boundaries of the normal physiological range exhibited no abrupt gel-liquid phase transitions when the lipids were analyzed as liposomes. Lipid packing and adaptational fatty acid composition responses are clearly influenced by differences in the temperature-salinity regime, which are reflected in overall cell function characteristics, such as the growth rate and the normal physiological range for growth. PMID- 10831421 TI - Sulfate-reducing bacteria methylate mercury at variable rates in pure culture and in marine sediments. AB - Differences in methylmercury (CH(3)Hg) production normalized to the sulfate reduction rate (SRR) in various species of sulfate-reducing bacteria (SRB) were quantified in pure cultures and in marine sediment slurries in order to determine if SRB strains which differ phylogenetically methylate mercury (Hg) at similar rates. Cultures representing five genera of the SRB (Desulfovibrio desulfuricans, Desulfobulbus propionicus, Desulfococcus multivorans, Desulfobacter sp. strain BG 8, and Desulfobacterium sp. strain BG-33) were grown in a strictly anoxic, minimal medium that received a dose of inorganic Hg 120 h after inoculation. The mercury methylation rates (MMR) normalized per cell were up to 3 orders of magnitude higher in pure cultures of members of SRB groups capable of acetate utilization (e.g., the family Desulfobacteriaceae) than in pure cultures of members of groups that are not able to use acetate (e.g., the family Desulfovibrionaceae). Little or no Hg methylation was observed in cultures of Desulfobacterium or Desulfovibrio strains in the absence of sulfate, indicating that Hg methylation was coupled to respiration in these strains. Mercury methylation, sulfate reduction, and the identities of sulfate-reducing bacteria in marine sediment slurries were also studied. Sulfate-reducing consortia were identified by using group-specific oligonucleotide probes that targeted the 16S rRNA molecule. Acetate-amended slurries, which were dominated by members of the Desulfobacterium and Desulfobacter groups, exhibited a pronounced ability to methylate Hg when the MMR were normalized to the SRR, while lactate-amended and control slurries had normalized MMR that were not statistically different. Collectively, the results of pure-culture and amended-sediment experiments suggest that members of the family Desulfobacteriaceae have a greater potential to methylate Hg than members of the family Desulfovibrionaceae have when the MMR are normalized to the SRR. Hg methylation potential may be related to genetic composition and/or carbon metabolism in the SRB. Furthermore, we found that in marine sediments that are rich in organic matter and dissolved sulfide rapid CH(3)Hg accumulation is coupled to rapid sulfate reduction. The observations described above have broad implications for understanding the control of CH(3)Hg formation and for developing remediation strategies for Hg-contaminated sediments. PMID- 10831422 TI - Characterization of a salt-tolerant family 42 beta-galactosidase from a psychrophilic antarctic Planococcus isolate. AB - We isolated a gram-positive, halotolerant psychrophile from a hypersaline pond located on the McMurdo Ice Shelf in Antarctica. A phylogenetic analysis of the 16S rRNA gene sequence of this organism showed that it is a member of the genus Planococcus. This assignment is consistent with the morphology and physiological characteristics of the organism. A gene encoding a beta-galactosidase in this isolate was cloned in an Escherichia coli host. Sequence analysis of this gene placed it in glycosidase family 42 most closely related to an enzyme from Bacillus circulans. Even though an increasing number of family 42 glycosidase sequences are appearing in databases, little information about the biochemical features of these enzymes is available. Therefore, we purified and characterized this enzyme. The purified enzyme did not appear to have any metal requirement, had an optimum pH of 6.5 and an optimum temperature of activity at 42 degrees C, and was irreversibly inactivated within 10 min when it was incubated at 55 degrees C. The enzyme had an apparent K(m) of 4.9 micromol of o-nitrophenyl-beta D-galactopyranoside, and the V(max) was 467 micromol of o-nitrophenol produced/min/mg of protein at 39 degrees C. Of special interest was the finding that the enzyme remained active at high salt concentrations, which makes it a possible reporter enzyme for halotolerant and halophilic organisms. PMID- 10831423 TI - Higher diversity of Rhizobium leguminosarum biovar viciae populations in arable soils than in grass soils. AB - The bacterial genetic diversity after long-term arable cultivation was compared with that under permanent grassland using replicated paired contrasts. Pea nodulating Rhizobium leguminosarum populations were sampled from pairs of arable and grass sites at four locations in Yorkshire, United Kingdom. Isolates were characterized using both chromosomal (16S-23S ribosomal DNA internal transcribed spacer PCR-restriction fragment length polymorphism) and plasmid (group-specific repC PCR amplification) markers. The diversities of chromosomal types, repC profiles, and combined genotypes were calculated using richness in types (adjusted to equal sample sizes by rarefaction), Shannon-Wiener index, and Simpson's index. The relative differences in diversity within each pair of sites were similar for all three diversity measures. Chromosomal types, repC profiles, and combined genotypes were each more diverse in arable soils than in grass soils at two of the four locations. The other comparisons showed no significant differences. We conclude that rhizobial diversity can be affected by differences between these two management regimens. Multiple regression analyses indicated that lower diversity was associated with high potential nitrogen and phosphate levels or with acidity. PMID- 10831424 TI - Effect of electron donor and solution chemistry on products of dissimilatory reduction of technetium by Shewanella putrefaciens. AB - To help provide a fundamental basis for use of microbial dissimilatory reduction processes in separating or immobilizing (99)Tc in waste or groundwaters, the effects of electron donor and the presence of the bicarbonate ion on the rate and extent of pertechnetate ion [Tc(VII)O(4)(-)] enzymatic reduction by the subsurface metal-reducing bacterium Shewanella putrefaciens CN32 were determined, and the forms of aqueous and solid-phase reduction products were evaluated through a combination of high-resolution transmission electron microscopy, X-ray absorption spectroscopy, and thermodynamic calculations. When H(2) served as the electron donor, dissolved Tc(VII) was rapidly reduced to amorphous Tc(IV) hydrous oxide, which was largely associated with the cell in unbuffered 0. 85% NaCl and with extracellular particulates (0.2 to 0.001 microm) in bicarbonate buffer. Cell associated Tc was present principally in the periplasm and outside the outer membrane. The reduction rate was much lower when lactate was the electron donor, with extracellular Tc(IV) hydrous oxide the dominant solid-phase reduction product, but in bicarbonate systems much less Tc(IV) was associated directly with the cell and solid-phase Tc(IV) carbonate may have been present. In the presence of carbonate, soluble (<0.001 microm) electronegative, Tc(IV) carbonate complexes were also formed that exceeded Tc(VII)O(4)(-) in electrophoretic mobility. Thermodynamic calculations indicate that the dominant reduced Tc species identified in the experiments would be stable over a range of E(h) and pH conditions typical of natural waters. Thus, carbonate complexes may represent an important pathway for Tc transport in anaerobic subsurface environments, where it has generally been assumed that Tc mobility is controlled by low-solubility Tc(IV) hydrous oxide and adsorptive, aqueous Tc(IV) hydrolysis products. PMID- 10831425 TI - Cellulose catabolism by Clostridium cellulolyticum growing in batch culture on defined medium. AB - A reinvestigation of cellulose degradation by Clostridium cellulolyticum in a bioreactor with pH control of the batch culture and using a defined medium was performed. Depending on cellulose concentration, the carbon flow distribution was affected, showing the high flexibility of the metabolism. With less than 6.7 g of cellulose liter(-1), acetate, ethanol, H(2), and CO(2) were the main end products of the fermentation and cellulose degradation reached more than 85% in 5 days. The electron flow from the glycolysis was balanced by the production of H(2) and ethanol, the latter increasing with increasing initial cellulose concentration. From 6.7 to 29.1 g of cellulose liter(-1), the percentage of cellulose degradation declined; most of the cellulase activity remained on the cellulose fibers, the maximum cell density leveled off, and the carbon flow was reoriented from ethanol to acetate. In addition to that of previously indicated end products, lactate production rose, and, surprisingly enough, pyruvate overflow occurred. Concomitantly the molar growth yield and the energetic yield of the biomass decreased. Growth arrest may be linked to sufficiently high carbon flow, leading to the accumulation of an intracellular inhibitory compound(s), as observed on cellobiose (E. Guedon, M. Desvaux, S. Payot, and H. Petitdemange, Microbiology 145:1831-1838, 1999). These results indicated that bacterial metabolism exhibited on cellobiose was distorted compared to that exhibited on a substrate more closely related to the natural ecosystem of C. cellulolyticum. To overcome growth arrest and to improve degradation at high cellulose concentrations (29.1 g liter(-1)), a reinoculation mode was evaluated. This procedure resulted in an increase in the maximum dry weight of cells (2,175 mg liter(-1)), cellulose solubilization (95%), and end product concentrations compared to a classical batch fermentation with a final dry weight of cells of 580 mg liter(-1) and 45% cellulose degradation within 18 days. PMID- 10831426 TI - Influence of cadmium and mercury on activities of ligninolytic enzymes and degradation of polycyclic aromatic hydrocarbons by Pleurotus ostreatus in soil. AB - The white-rot fungus Pleurotus ostreatus was able to degrade the polycyclic aromatic hydrocarbons (PAHs) benzo[a]anthracene, chrysene, benzo[b]fluoranthene, benzo[k]fluoranthene, benzo[a]pyrene, dibenzo[a,h]anthracene, and benzo[ghi]perylene in nonsterile soil both in the presence and in the absence of cadmium and mercury. During 15 weeks of incubation, recovery of individual compounds was 16 to 69% in soil without additional metal. While soil microflora contributed mostly to degradation of pyrene (82%) and benzo[a]anthracene (41%), the fungus enhanced the disappearance of less-soluble polycyclic aromatic compounds containing five or six aromatic rings. Although the heavy metals in the soil affected the activity of ligninolytic enzymes produced by the fungus (laccase and Mn-dependent peroxidase), no decrease in PAH degradation was found in soil containing Cd or Hg at 10 to 100 ppm. In the presence of cadmium at 500 ppm in soil, degradation of PAHs by soil microflora was not affected whereas the contribution of fungus was negligible, probably due to the absence of Mn dependent peroxidase activity. In the presence of Hg at 50 to 100 ppm or Cd at 100 to 500 ppm, the extent of soil colonization by the fungus was limited. PMID- 10831427 TI - Hydroxylated metabolites of 2,4-dichlorophenol imply a fenton-type reaction in Gloeophyllum striatum. AB - While degrading 2,4-dichlorophenol, two strains of Gloeophyllum striatum, a basidiomycetous fungus causing brown rot decay of wood, simultaneously produced 4 chlorocatechol and 3,5-dichlorocatechol. These metabolites were identified by comparing high-performance liquid chromatography retention times and mass spectral data with those of chemically synthesized standards. Under similar conditions, 3-hydroxyphthalic hydrazide was generated from phthalic hydrazide, a reaction assumed to indicate hydroxyl radical formation. Accordingly, during chemical degradation of 2,4-dichlorophenol by Fenton's reagent, identical metabolites were formed. Both activities, the conversion of 2,4-[U (14)C]dichlorophenol into (14)CO(2) and the generation of 3-hydroxyphthalic hydrazide, were strongly inhibited by the hydroxyl radical scavenger mannitol and in the absence of iron. These results provide new evidence in favor of a Fenton type degradation mechanism operative in Gloeophyllum. PMID- 10831428 TI - Characterization of a bifunctional enzyme fusion of trehalose-6-phosphate synthetase and trehalose-6-phosphate phosphatase of Escherichia coli. AB - To test the effect of the physical proximity of two enzymes catalyzing sequential reactions, a bifunctional fusion enzyme, TPSP, was constructed by fusing the Escherichia coli genes for trehalose-6-phosphate (T6P) synthetase (TPS) and trehalose-6-phosphate phosphatase (TPP). TPSP catalyzes the sequential reaction in which T6P is formed and then dephosphorylated, leading to the synthesis of trehalose. The fused chimeric gene was overexpressed in E. coli and purified to near homogeneity; its molecular weight was 88,300, as expected. The K(m) values of the TPSP fusion enzyme for the sequential overall reaction from UDP-glucose and glucose 6-phosphate to trehalose were smaller than those of an equimolar mixture of TPS and TPP (TPS/TPP). However, the k(cat) values of TPSP were similar to those of TPS/TPP, resulting in a 3.5- to 4.0-fold increase in the catalytic efficiency (k(cat)/K(m)). The K(m) and k(cat) values of TPSP and TPP for the phosphatase reaction from T6P to trehalose were quite similar. This suggests that the increased catalytic efficiency results from the proximity of TPS and TPP in the TPSP fusion enzyme. The thermal stability of the TPSP fusion enzyme was quite similar to that of the TPS/TPP mixture, suggesting that the structure of each enzyme moiety in TPSP is unperturbed by intramolecular constraint. These results clearly demonstrate that the bifunctional fusion enzyme TPSP catalyzing sequential reactions has kinetic advantages over a mixture of both enzymes (TPS and TPP). These results are also supported by the in vivo accumulation of up to 0.48 mg of trehalose per g of cells after isopropyl-beta-D-thiogalactopyranoside treatment of cells harboring the construct encoding TPSP. PMID- 10831429 TI - Isolation and characterization of strains CVO and FWKO B, two novel nitrate reducing, sulfide-oxidizing bacteria isolated from oil field brine. AB - Bacterial strains CVO and FWKO B were isolated from produced brine at the Coleville oil field in Saskatchewan, Canada. Both strains are obligate chemolithotrophs, with hydrogen, formate, and sulfide serving as the only known energy sources for FWKO B, whereas sulfide and elemental sulfur are the only known electron donors for CVO. Neither strain uses thiosulfate as an energy source. Both strains are microaerophiles (1% O(2)). In addition, CVO grows by denitrification of nitrate or nitrite whereas FWKO B reduces nitrate only to nitrite. Elemental sulfur is the sole product of sulfide oxidation by FWKO B, while CVO produces either elemental sulfur or sulfate, depending on the initial concentration of sulfide. Both strains are capable of growth under strictly autotrophic conditions, but CVO uses acetate as well as CO(2) as its sole carbon source. Neither strain reduces sulfate; however, FWKO B reduces sulfur and displays chemolithoautotrophic growth in the presence of elemental sulfur, hydrogen, and CO(2). Both strains grow at temperatures between 5 and 40 degrees C. CVO is capable of growth at NaCl concentrations as high as 7%. The present 16s rRNA analysis suggests that both strains are members of the epsilon subdivision of the division Proteobacteria, with CVO most closely related to Thiomicrospira denitrifcans and FWKO B most closely related to members of the genus Arcobacter. The isolation of these two novel chemolithotrophic sulfur bacteria from oil field brine suggests the presence of a subterranean sulfur cycle driven entirely by hydrogen, carbon dioxide, and nitrate. PMID- 10831430 TI - Bile salt hydrolase of Bifidobacterium longum-biochemical and genetic characterization. AB - A bile salt hydrolase (BSH) was isolated from Bifidobacterium longum SBT2928, purified, and characterized. Furthermore, we describe for the first time cloning and analysis of the gene encoding BSH (bsh) in a member of the genus Bifidobacterium. The enzyme has a native molecular weight of 125,000 to 130,000 and a subunit molecular weight of 35,024, as determined from the deduced amino acid sequence, indicating that the enzyme is a tetramer. The pH optimum of B. longum BSH is between 5 and 7, and the temperature optimum is 40 degrees C. The enzyme is strongly inhibited by thiol enzyme inhibitors, indicating that a Cys residue is likely to be involved in the catalytic reaction. The BSH of B. longum can hydrolyze all six major human bile salts and at least two animal bile salts. A slight preference for glycine-conjugated bile acids was detected based on both the specificity and the K(m) values. The nucleotide sequence of bsh was determined and used for homology studies, transcript analysis, and construction and analysis of various mutants. The levels of homology with BSH of other bacteria and with penicillin V acylase (PVA) of Bacillus sphaericus were high. On the basis of the similarity of BSH and PVA, whose crystal structure has been elucidated, BSH can be classified as an N-terminal nucleophile hydrolase with Cys as the N-terminal amino acid. This classification was confirmed by the fact that a Cys1Ala exchange by site-directed mutagenesis resulted in an inactive protein. Reverse transcription-PCR experiments revealed that bsh is part of an operon containing at least two genes, bsh and glnE (GlnE is glutamine synthetase adenylyltransferase). Two UV-induced BSH-negative mutants and one spontaneous BSH negative mutant were isolated from B. longum SBT2928 cultures and characterized. These mutants had point mutations that inactivated bsh by premature termination, frameshift, or amino acid exchange. PMID- 10831431 TI - Restriction-site-specific PCR as a rapid test to detect enterohemorrhagic Escherichia coli O157:H7 strains in environmental samples. AB - Enterohemorrhagic Escherichia coli (EHEC) O157:H7 is an important food-borne pathogen in industrialized countries. We developed a rapid and simple test for detecting E. coli O157:H7 using a method based on restriction site polymorphisms. Restriction-site-specific PCR (RSS-PCR) involves the amplification of DNA fragments using primers based on specific restriction enzyme recognition sequences, without the use of endonucleases, to generate a set of amplicons that yield "fingerprint" patterns when resolved electrophoretically on an agarose gel. The method was evaluated in a blinded study of E. coli isolates obtained from environmental samples collected at beef cattle feedyards. The 54 isolates were all initially identified by a commonly used polyclonal antibody test as belonging to O157:H7 serotype. They were retested by anti-O157 and anti-H7 monoclonal antibody enzyme-linked immunosorbent assay (ELISA). The RSS-PCR method identified all 28 isolates that were shown to be E. coli O157:H7 by the monoclonal antibody ELISA as belonging to the O157:H7 serotype. Of the remaining 26 ELISA-confirmed non-O157:H7 strains, the method classified 25 strains as non-O157:H7. The specificity of the RSS-PCR results correlated better with the monoclonal antibody ELISA than with the polyclonal antibody latex agglutination tests. The RSS-PCR method may be a useful test to distinguish E. coli O157:H7 from a large number of E. coli isolates from environmental samples. PMID- 10831432 TI - Detection of astroviruses, enteroviruses, and adenovirus types 40 and 41 in surface waters collected and evaluated by the information collection rule and an integrated cell culture-nested PCR procedure. AB - We evaluated the use of an integrated cell culture-reverse transcription-PCR (ICC RT-PCR) procedure coupled with nested PCR to detect human astroviruses, enteroviruses, and adenovirus types 40 and 41 in surface water samples that were collected and evaluated by using the Information Collection Rule (ICR) method. The results obtained with the ICC-RT-PCR-nested PCR method were compared to the results obtained with the total culturable virus assay-most-probable-number (TCVA MPN) method, the method recommended by the U.S. Environmental Protection Agency for monitoring viruses in surface and finished waters. Twenty-nine ICR surface water samples were analyzed. Viruses were concentrated by using filter adsorption beef extract elution and organic flocculation techniques, and then the preparations were evaluated for viruses by visualizing cytopathic effects in the Buffalo green monkey kidney (BGMK) cell line. In the ICC-RT-PCR-nested PCR technique we used Caco-2 cells to propagate astroviruses and enteroviruses (ICC step), and we used BGMK cells to propagate adenovirus types 40 and 41, as well as enteroviruses. Fifteen of the 29 samples (51.7%) were positive for astrovirus as determined by the ICC-RT-PCR-nested PCR method, and eight of these samples (27.5%) contained infectious astrovirus. Seventeen of the 29 samples (58.6%) were positive for enteroviruses when the BGMK cell line was used, and six (27.6%) of these samples were determined to be infectious. Fourteen of the 29 samples (48.3%) were positive for adenovirus types 40 and 41, and 11 (37.9%) of these samples were determined to be infectious. Twenty-seven of the 29 samples (93.1%) were positive for a virus, and 19 (68.9%) of the samples were positive for an infectious virus. Only 5 of the 29 samples (17.2%) were positive as determined by the TCVA-MPN method. The ICC-RT-PCR-nested PCR method provided increased sensitivity compared to the TCVA-MPN method. PMID- 10831433 TI - Effects of metal phytoextraction practices on the indigenous community of arbuscular mycorrhizal fungi at a metal-contaminated landfill. AB - Phytoextraction involves use of plants to remove toxic metals from soil. We examined the effects of phytoextraction practices with three plant species (Silene vulgaris, Thlaspi caerulescens, and Zea mays) and a factorial variation of soil amendments (either an ammonium or nitrate source of nitrogen and the presence or absence of an elemental sulfur supplement) on arbuscular mycorrhizal (AM) fungi (Glomales, Zygomycetes) at a moderately metal-contaminated landfill located in St. Paul, Minn. Specifically, we tested whether the applied treatments affected the density of glomalean spores and AM root colonization in maize. Glomalean fungi from the landfill were grouped into two morphotypes characterized by either light-colored spores (LCS) or dark-colored spores (DCS). Dominant species of the LCS morphotype were Glomus mosseae and an unidentified Glomus sp., whereas the DCS morphotype was dominated by Glomus constrictum. The density of spores of the LCS morphotype from the phytoremediated area was lower than the density of these spores in the untreated landfill soil. Within the experimental area, spore density of the LCS morphotype in the rhizosphere of mycorrhizal maize was significantly higher than in rhizospheres of nonmycorrhizal S. vulgaris or T. caerulescens. Sulfur supplement increased vesicular root colonization in maize and exerted a negative effect on spore density in maize rhizosphere. We conclude that phytoextraction practices, e.g., the choice of plant species and soil amendments, may have a great impact on the quantity and species composition of glomalean propagules as well as on mycorrhiza functioning during long-term metal remediation treatments. PMID- 10831434 TI - Molecular cloning, characterization, and differential expression of a glucoamylase gene from the basidiomycetous fungus Lentinula edodes. AB - The complete nucleotide sequence of putative glucoamylase gene gla1 from the basidiomycetous fungus Lentinula edodes strain L54 is reported. The coding region of the genomic glucoamylase sequence, which is preceded by eukaryotic promoter elements CAAT and TATA, spans 2,076 bp. The gla1 gene sequence codes for a putative polypeptide of 571 amino acids and is interrupted by seven introns. The open reading frame sequence of the gla1 gene shows strong homology with those of other fungal glucoamylase genes and encodes a protein with an N-terminal catalytic domain and a C-terminal starch-binding domain. The similarity between the Gla1 protein and other fungal glucoamylases is from 45 to 61%, with the region of highest conservation found in catalytic domains and starch-binding domains. We compared the kinetics of glucoamylase activity and levels of gene expression in L. edodes strain L54 grown on different carbon sources (glucose, starch, cellulose, and potato extract) and in various developmental stages (mycelium growth, primordium appearance, and fruiting body formation). Quantitative reverse transcription PCR utilizing pairs of primers specific for gla1 gene expression shows that expression of gla1 was induced by starch and increased during the process of fruiting body formation, which indicates that glucoamylases may play an important role in the morphogenesis of the basidiomycetous fungus. PMID- 10831435 TI - Role of volatile fatty acids in development of the cecal microflora in broiler chickens during growth. AB - It is known that volatile fatty acids can inhibit growth of species of the family Enterobacteriaceae in vitro. However, whether these volatile fatty acids affect bacterial populations in the ceca of chickens is unknown. Therefore, a study was conducted to investigate if changes in volatile fatty acids in ceca of broiler chickens during growth affect bacterial populations. Results showed that members of the Enterobacteriaceae and enterococci are present in large numbers in 3-day old broilers and start to decrease when broilers grow older. Lactobacilli are present in large numbers as well in 3-day-old broilers, but they remain stable during the growth of broilers. Acetate, butyrate, and propionate increase from undetectable levels in 1-day-old broilers to high concentrations in 15-day-old broilers, after which they stabilize. Significant negative correlations could be calculated between numbers of Enterobacteriaceae and concentrations of undissociated acetate, propionate, and butyrate. Furthermore, pure cultures of Enterobacteriaceae isolated from the ceca were grown in the presence of volatile fatty acids. Growth rates and maximal optical density decreased when these strains grew in the presence of increasing volatile fatty acid concentrations. It is concluded that volatile fatty acids are responsible for the reduction in numbers of Enterobacteriaceae in the ceca of broiler chickens during growth. PMID- 10831437 TI - Surface of lactic acid bacteria: relationships between chemical composition and physicochemical properties. AB - The surface chemical composition and physicochemical properties (hydrophobicity and zeta potential) of two lactic acid bacteria, Lactococcus lactis subsp. lactis bv. diacetilactis and Lactobacillus helveticus, have been investigated using cells harvested in exponential or stationary growth phase. The surface composition determined by X-ray photoelectron spectroscopy (XPS) was converted into a molecular composition in terms of proteins, polysaccharides, and hydrocarbonlike compounds. The concentration of the last was always below 15% (wt/wt), which is related to the hydrophilic character revealed by water contact angles of less than 30 degrees. The surfaces of L. lactis cells had a polysaccharide concentration about twice that of proteins. The S-layer of L. helveticus was either interrupted or crossed by polysaccharide-rich compounds; the concentration of the latter was higher in the stationary growth phase than in the exponential growth phase. Further progress was made in the interpretation of XPS data in terms of chemical functions by showing that the oxygen component at 531.2 eV contains a contribution of phosphate in addition to the main contribution of the peptide link. The isoelectric points were around 2 and 3, and the electrophoretic mobilities above pH 5 (ionic strength, 1 mM) were about -3.0 x 10(-8) and -0.6 x 10(-8) m(2) s(-1) V(-1) for L. lactis and L. helveticus, respectively. The electrokinetic properties of the latter reveal the influence of carboxyl groups, while the difference between the two strains is related to a difference between N/P surface concentration ratios, reflecting the relative exposure of proteins and phosphate groups at the surface. PMID- 10831436 TI - Cloning the soil metagenome: a strategy for accessing the genetic and functional diversity of uncultured microorganisms. AB - Recent progress in molecular microbial ecology has revealed that traditional culturing methods fail to represent the scope of microbial diversity in nature, since only a small proportion of viable microorganisms in a sample are recovered by culturing techniques. To develop methods to investigate the full extent of microbial diversity, we used a bacterial artificial chromosome (BAC) vector to construct libraries of genomic DNA isolated directly from soil (termed metagenomic libraries). To date, we have constructed two such libraries, which contain more than 1 Gbp of DNA. Phylogenetic analysis of 16S rRNA gene sequences recovered from one of the libraries indicates that the BAC libraries contain DNA from a wide diversity of microbial phyla, including sequences from diverse taxa such as the low-G+C, gram-positive Acidobacterium, Cytophagales, and Proteobacteria. Initial screening of the libraries in Escherichia coli identified several clones that express heterologous genes from the inserts, confirming that the BAC vector can be used to maintain, express, and analyze environmental DNA. The phenotypes expressed by these clones include antibacterial, lipase, amylase, nuclease, and hemolytic activities. Metagenomic libraries are a powerful tool for exploring soil microbial diversity, providing access to the genetic information of uncultured soil microorganisms. Such libraries will be the basis of new initiatives to conduct genomic studies that link phylogenetic and functional information about the microbiota of environments dominated by microorganisms that are refractory to cultivation. PMID- 10831438 TI - Use of randomly amplified polymorphic DNA as a means of developing genus- and strain-specific Streptomyces DNA probes. AB - We have analyzed 20 randomly amplified polymorphic DNA (RAPD) primers against 36 Streptomyces strains, including 17 taxonomically undefined strains, 25 nonstreptomycete actinomycetes, and 12 outgroups consisting of gram-positive and negative species. Most of the primers were useful in identifying unique DNA polymorphisms of all strains tested. We have used RAPD techniques to develop a genus-specific probe, one not necessarily targeting the ribosomal gene, for Streptomyces, and a strain-specific probe for the biological control agent Streptomyces lydicus WYEC108. In the course of these investigations, small-scale DNA isolations were also developed for efficiently isolating actinomycete DNA. Various modifications of isolation procedures for soil DNA were compared, and the reliability and specificity of the RAPD methodology were tested by specifically detecting the S. lydicus WYEC108 in DNA isolated from soil. PMID- 10831439 TI - Biosynthesis of the lantibiotic mersacidin: organization of a type B lantibiotic gene cluster. AB - The biosynthetic gene cluster (12.3 kb) of mersacidin, a lanthionine-containing antimicrobial peptide, is located on the chromosome of the producer, Bacillus sp. strain HIL Y-85,54728 in a region that corresponds to 348 degrees on the chromosome of Bacillus subtilis 168. It consists of 10 open reading frames and contains, in addition to the previously described mersacidin structural gene mrsA (G. Bierbaum, H. Brotz, K.-P. Koller, and H.-G. Sahl, FEMS Microbiol. Lett. 127:121-126, 1995), two genes, mrsM and mrsD, coding for enzymes involved in posttranslational modification of the prepeptide; one gene, mrsT, coding for a transporter with an associated protease domain; and three genes, mrsF, mrsG, and mrsE, encoding a group B ABC transporter that could be involved in producer self protection. Additionally, three regulatory genes are part of the gene cluster, i.e., mrsR2 and mrsK2, which encode a two-component regulatory system which seems to be necessary for the transcription of the mrsFGE operon, and mrsR1, which encodes a protein with similarity to response regulators. Transcription of mrsA sets in at early stationary phase (between 8 and 16 h of culture). PMID- 10831440 TI - Use of repetitive DNA sequences and the PCR To differentiate Escherichia coli isolates from human and animal sources. AB - The rep-PCR DNA fingerprint technique, which uses repetitive intergenic DNA sequences, was investigated as a way to differentiate between human and animal sources of fecal pollution. BOX and REP primers were used to generate DNA fingerprints from Escherichia coli strains isolated from human and animal sources (geese, ducks, cows, pigs, chickens, and sheep). Our initial studies revealed that the DNA fingerprints obtained with the BOX primer were more effective for grouping E. coli strains than the DNA fingerprints obtained with REP primers. The BOX primer DNA fingerprints of 154 E. coli isolates were analyzed by using the Jaccard band-matching algorithm. Jackknife analysis of the resulting similarity coefficients revealed that 100% of the chicken and cow isolates and between 78 and 90% of the human, goose, duck, pig, and sheep isolates were assigned to the correct source groups. A dendrogram constructed by using Jaccard similarity coefficients almost completely separated the human isolates from the nonhuman isolates. Multivariate analysis of variance, a form of discriminant analysis, successfully differentiated the isolates and placed them in the appropriate source groups. Taken together, our results indicate that rep-PCR performed with the BOX A1R primer may be a useful and effective tool for rapidly determining sources of fecal pollution. PMID- 10831441 TI - Analysis of the fecal microflora of human subjects consuming a probiotic product containing Lactobacillus rhamnosus DR20. AB - The composition of the fecal microflora of 10 healthy subjects was monitored before (6-month control period), during (6-month test period), and after (3-month posttest period) the administration of a milk product containing Lactobacillus rhamnosus DR20 (daily dose, 1.6 x 10(9) lactobacilli). Monthly fecal samples were examined by a variety of methods, including bacteriological culture analysis, fluorescent in situ hybridization with group-specific DNA probes, denaturing gradient gel electrophoresis of the V2-V3 region of 16S rRNA genes amplified by PCR, gas-liquid chromatography, and bacterial enzyme activity analysis. The composition of the Lactobacillus population of each subject was analyzed by pulsed-field gel electrophoresis of bacterial DNA digests in order to differentiate between DR20 and other strains present in the samples. Representative isolates of lactobacilli were identified to the species level by sequencing the V2-V3 region of their 16S rRNA genes and comparing the sequences obtained (BLAST search) to sequences in the GenBank database. DR20 was detected in the feces of all of the subjects during the test period, but at different frequencies. The presence of DR20 among the numerically predominant strains was related to the presence or absence of a stable indigenous population of lactobacilli during the control period. Strain DR20 did not persist at levels of >10(2) cells per g in the feces of most of the subjects after consumption of the product ceased; the only exception was one subject in which this strain was detected for 2 months during the posttest period. We concluded that consumption of the DR20-containing milk product transiently altered the Lactobacillus and enterococcal contents of the feces of the majority of consumers without markedly affecting biochemical or other bacteriological factors. PMID- 10831442 TI - Functional exoenzymes as indicators of metabolically active bacteria in 124,000 year-Old sapropel layers of the eastern Mediterranean Sea. AB - Hydrolytic exoenzymes as indicators of metabolically active bacteria were investigated in four consecutive sapropel layers collected from bathyal sediments of the eastern Mediterranean Sea. For comparison, the organic carbon-poor layers between the sapropels, sediment from the anoxic Urania basin, and sediments of intertidal mud flats of the German Wadden Sea were also analyzed. The sapropel layers contained up to 1.5. 10(8) bacterial cells cm(-3), whereas cell numbers in the intermediate layers were lower by a factor of 10. In sapropels, the determination of exoenzyme activity with fluorescently labeled substrate analogues was impaired by the strong adsorption of up to 97% of the enzymatically liberated fluorophores (4-methylumbelliferone [MUF] and 7-amino-4-methylcoumarin [MCA]) to the sediment particles. Because all established methods for the extraction of adsorbed fluorophores proved to be inadequate for sapropel sediments, we introduce a correction method which is based on the measurement of equilibrium adsorption isotherms for both compounds. Using this new approach, high activities of aminopeptidase and alkaline phosphatase were detected even in a 124,000-year-old sapropel layer, whereas the activity of beta-glucosidase was low in all layers. So far, it had been assumed that the organic matter which constitutes the sapropels is highly refractory. The high potential activities of bacterial exoenzymes indicate that bacteria in Mediterranean sapropels are metabolically active and utilize part of the subfossil kerogen. Since a high adsorption capacity was determined not only for the low-molecular-weight compounds MUF and MCA but also for DNA, the extraordinarily strong adsorption of structurally different substrates to the sapropel matrix appears to be the major reason for the long-term preservation of biodegradable carbon in this environment. PMID- 10831443 TI - Generation of food-grade recombinant lactic acid bacterium strains by site specific recombination. AB - The construction of a delivery and clearing system for the generation of food grade recombinant lactic acid bacterium strains, based on the use of an integrase (Int) and a resolvo-invertase (beta-recombinase) and their respective target sites (attP-attB and six, respectively) is reported. The delivery system contains a heterologous replication origin and antibiotic resistance markers surrounded by two directly oriented six sites, a multiple cloning site where passenger DNA could be inserted (e.g., the cI gene of bacteriophage A2), the int gene, and the attP site of phage A2. The clearing system provides a plasmid-borne gene encoding beta-recombinase. The nonreplicative vector-borne delivery system was transformed into Lactobacillus casei ATCC 393 and, by site-specific recombination, integrated as a single copy in an orientation- and Int-dependent manner into the attB site present in the genome of the host strain. The transfer of the clearing system into this strain, with the subsequent expression of the beta-recombinase, led to site-specific DNA resolution of the non-food-grade DNA. These methods were validated by the construction of a stable food-grade L. casei ATCC 393-derived strain completely immune to phage A2 infection during milk fermentation. PMID- 10831444 TI - Comparative survival rates of human-derived probiotic Lactobacillus paracasei and L. salivarius strains during heat treatment and spray drying. AB - Spray drying of skim milk was evaluated as a means of preserving Lactobacillus paracasei NFBC 338 and Lactobacillus salivarius UCC 118, which are human-derived strains with probiotic potential. Our initial experiments revealed that NFBC 338 is considerably more heat resistant in 20% (wt/vol) skim milk than UCC 118 is; the comparable decimal reduction times were 11.1 and 1.1 min, respectively, at 59 degrees C. An air outlet temperature of 80 to 85 degrees C was optimal for spray drying; these conditions resulted in powders with moisture contents of 4.1 to 4.2% and viable counts of 3.2 x 10(9) CFU/g for NFBC 338 and 5.2 x 10(7) CFU/g for UCC 118. Thus, L. paracasei NFBC 338 survived better than L. salivarius UCC 118 during spray drying; similar results were obtained when we used confocal scanning laser microscopy and LIVE/DEAD BacLight viability staining. In addition, confocal scanning laser microscopy revealed that the probiotic lactobacilli were located primarily in the powder particles. Although both spray-dried cultures appeared to be stressed, as shown by increased sensitivity to NaCl, bacteriocin production by UCC 118 was not affected by the process, nor was the activity of the bacteriocin peptide. The level of survival of NFBC 338 remained constant at approximately 1 x 10(9) CFU/g during 2 months of powder storage at 4 degrees C, while a decline in the level of survival of approximately 1 log (from 7.2 x 10(7) to 9.5 x 10(6) CFU/g) was observed for UCC 118 stored under the same conditions. However, survival of both Lactobacillus strains during powder storage was inversely related to the storage temperature. Our data demonstrate that spray drying may be a cost-effective way to produce large quantities of some probiotic cultures. PMID- 10831445 TI - Phylogenetic diversity among geographically dispersed Chlamydiales endosymbionts recovered from clinical and environmental isolates of Acanthamoeba spp. AB - The recently proposed reorganization of the order Chlamydiales and description of new taxa are broadening our perception of this once narrowly defined taxon. We have recovered four strains of gram-negative cocci endosymbiotic in Acanthamoeba spp., representing 5% of the Acanthamoeba sp. isolates examined, which displayed developmental life cycles typical of members of the Chlamydiales. One of these endosymbiont strains was found stably infecting an amoebic isolate recovered from a case of amoebic keratitis in North America, with three others found in acanthamoebae recovered from environmental sources in North America (two isolates) and Europe (one isolate). Analyses of nearly full-length 16S rRNA gene sequences of these isolates by neighbor joining, parsimony, and distance matrix methods revealed their clustering with other members of the Chlamydiales but in a lineage separate from those of the genera Chlamydia, Chlamydophila, Simkania, and Waddlia (sequence similarities, <88%) and including the recently described species Parachlamydia acanthamoebae (sequence similarities, 91.2 to 93.1%). With sequence similarities to each other of 91.4 to 99.4%, these four isolates of intra-amoebal endosymbionts may represent three distinct species and, perhaps, new genera within the recently proposed family Parachlamydiaceae. Fluorescently labeled oligonucleotide probes targeted to 16S rRNA signature regions were able to readily differentiate two groups of intra-amoebal endosymbionts which corresponded to two phylogenetic lineages. These results reveal significant phylogenetic diversity occurring among the Chlamydiales in nontraditional host species and supports the existence of a large environmental reservoir of related species. Considering that all described species of Chlamydiales are known to be pathogenic, further investigation of intra-amoebal parachlamydiae as disease producing agents is warranted. PMID- 10831446 TI - Physiologic determinants of radiation resistance in Deinococcus radiodurans. AB - Immense volumes of radioactive wastes, which were generated during nuclear weapons production, were disposed of directly in the ground during the Cold War, a period when national security priorities often surmounted concerns over the environment. The bacterium Deinococcus radiodurans is the most radiation resistant organism known and is currently being engineered for remediation of the toxic metal and organic components of these environmental wastes. Understanding the biotic potential of D. radiodurans and its global physiological integrity in nutritionally restricted radioactive environments is important in development of this organism for in situ bioremediation. We have previously shown that D. radiodurans can grow on rich medium in the presence of continuous radiation (6,000 rads/h) without lethality. In this study we developed a chemically defined minimal medium that can be used to analyze growth of this organism in the presence and in the absence of continuous radiation; whereas cell growth was not affected in the absence of radiation, cells did not grow and were killed in the presence of continuous radiation. Under nutrient-limiting conditions, DNA repair was found to be limited by the metabolic capabilities of D. radiodurans and not by any nutritionally induced defect in genetic repair. The results of our growth studies and analysis of the complete D. radiodurans genomic sequence support the hypothesis that there are several defects in D. radiodurans global metabolic regulation that limit carbon, nitrogen, and DNA metabolism. We identified key nutritional constituents that restore growth of D. radiodurans in nutritionally limiting radioactive environments. PMID- 10831447 TI - Bacillus anthracis, Bacillus cereus, and Bacillus thuringiensis--one species on the basis of genetic evidence. AB - Bacillus anthracis, Bacillus cereus, and Bacillus thuringiensis are members of the Bacillus cereus group of bacteria, demonstrating widely different phenotypes and pathological effects. B. anthracis causes the acute fatal disease anthrax and is a potential biological weapon due to its high toxicity. B. thuringiensis produces intracellular protein crystals toxic to a wide number of insect larvae and is the most commonly used biological pesticide worldwide. B. cereus is a probably ubiquitous soil bacterium and an opportunistic pathogen that is a common cause of food poisoning. In contrast to the differences in phenotypes, we show by multilocus enzyme electrophoresis and by sequence analysis of nine chromosomal genes that B. anthracis should be considered a lineage of B. cereus. This determination is not only a formal matter of taxonomy but may also have consequences with respect to virulence and the potential of horizontal gene transfer within the B. cereus group. PMID- 10831448 TI - Cloning, expression, and sequence analysis of the gene encoding the alkali stable, thermostable endoxylanase from alkalophilic, mesophilic Bacillus sp. Strain NG-27. AB - Alkalophilic Bacillus sp. strain NG-27 produces a 42-kDa endoxylanase active at 70 degrees C and at a pH of 8.4. The gene for this endoxylanase was cloned and sequenced. The gene contained one open reading frame of 1,215 bases. An active site characteristic of the family 10 beta-glycanases was recognized between amino acids 303 and 313, with the active glutamate at position 310. Though highly thermostable, the enzyme contains no cysteine residue. PMID- 10831449 TI - Effects of soil and water content on methyl bromide oxidation by the ammonia oxidizing bacterium Nitrosomonas europaea. AB - Little information exists on the potential of NH(3)-oxidizing bacteria to cooxidize halogenated hydrocarbons in soil. A study was conducted to examine the cooxidation of methyl bromide (MeBr) by an NH(3)-oxidizing bacterium, Nitrosomonas europaea, under soil conditions. Soil and its water content modified the availability of NH(4)(+) and MeBr and influenced the relative rates of substrate (NH(3)) and cosubstrate (MeBr) oxidations. These observations highlight the complexity associated with characterizing soil cooxidative activities when soil and water interact to differentially affect substrate and cosubstrate availabilities. PMID- 10831450 TI - Methods for intense aeration, growth, storage, and replication of bacterial strains in microtiter plates. AB - Miniaturized growth systems for heterogeneous culture collections are not only attractive in reducing demands for incubation space and medium but also in making the parallel handling of large numbers of strains more practicable. We report here on the optimization of oxygen transfer rates in deep-well microtiter plates and the development of a replication system allowing the simultaneous and reproducible sampling of 96 frozen glycerol stock cultures while the remaining culture volume remains frozen. Oxygen transfer rates were derived from growth curves of Pseudomonas putida and from rates of oxygen disappearance due to the cobalt-catalyzed oxidation of sulfite. Maximum oxygen transfer rates (38 mmol liter(-1) h(-1), corresponding to a mass transfer coefficient of 188 h(-1)) were measured during orbital shaking at 300 rpm at a shaking diameter of 5 cm and a culture volume of 0.5 ml. A shaking diameter of 2.5 cm resulted in threefold lower values. These high oxygen transfer rates allowed P. putida to reach a cell density of approximately 9 g (dry weight) liter(-1) during growth on a glucose mineral medium at culture volumes of up to 1 ml. The growth-and-replication system was evaluated for a culture collection consisting of aerobic strains, mainly from the genera Pseudomonas, Rhodococcus, and Alcaligenes, using mineral media and rich media. Cross-contamination and excessive evaporation during vigorous aeration were adequately prevented by the use of a sandwich cover of spongy silicone and cotton wool on top of the microtiter plates. PMID- 10831451 TI - Characterization of AbiR, a novel multicomponent abortive infection mechanism encoded by plasmid pKR223 of Lactococcus lactis subsp. lactis KR2. AB - The native lactococcal plasmid pKR223 encodes two distinct phage resistance mechanisms, a restriction and modification (R/M) system designated LlaKR2I and an abortive infection mechanism (Abi) which affects prolate-headed-phage proliferation. The nucleotide sequence of a 16,174-bp segment of pKR223 encompassing both the R/M and Abi determinants has been determined, and sequence analysis has validated the novelty of the Abi system, which has now been designated AbiR. Analysis of deletion and insertion clones demonstrated that AbiR was encoded by two genetic loci, separated by the LlaKR2I R/M genes. Mechanistic studies on the AbiR phenotype indicated that it was heat sensitive and that it impeded phage DNA replication. These data indicated that AbiR is a novel multicomponent, heat-sensitive, "early"-functioning Abi system and is the first lactococcal Abi system described which is encoded by two separated genetic loci. PMID- 10831452 TI - Characterization of metabolites during biodegradation of hexahydro-1, 3,5 trinitro-1,3,5-triazine (RDX) with municipal anaerobic sludge. AB - The biodegradation of hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) in liquid cultures with municipal anaerobic sludge showed that at least two degradation routes were involved in the disappearance of the cyclic nitramine. In one route, RDX was reduced to give the familiar nitroso derivatives hexahydro-1-nitroso-3,5 dinitro-1,3, 5-triazine (MNX) and hexahydro-1,3-dinitroso-5-nitro-1,3,5-triazine (DNX). In the second route, two novel metabolites, methylenedinitramine [(O(2)NNH)(2)CH(2)] and bis(hydroxymethyl)nitramine [(HOCH(2))(2)NNO(2)], formed and were presumed to be ring cleavage products produced by enzymatic hydrolysis of the inner C---N bonds of RDX. None of the above metabolites accumulated in the system, and they disappeared to produce nitrous oxide (N(2)O) as a nitrogen containing end product and formaldehyde (HCHO), methanol (MeOH), and formic acid (HCOOH) that in turn disappeared to produce CH(4) and CO(2) as carbon-containing end products. PMID- 10831453 TI - Rhizobitoxine production by Bradyrhizobium elkanii enhances nodulation and competitiveness on Macroptilium atropurpureum. AB - Application of 1-aminoocyclopropane-1-carboxylic acid, an ethylene precursor, decreased nodulation of Macroptilium atropurpureum by Bradyrhizobium elkanii. B. elkanii produces rhizobitoxine, an ethylene synthesis inhibitor. Elimination of rhizobitoxine production in B. elkanii increased ethylene evolution and decreased nodulation and competitiveness on M. atropurpureum. These results suggest that rhizobitoxine enhances nodulation and competitiveness of B. elkanii on M. atropurpureum. PMID- 10831454 TI - Microbiological transformation of enrofloxacin by the fungus Mucor ramannianus. AB - Enrofloxacin metabolism by Mucor ramannianus was investigated as a model for the biotransformation of veterinary fluoroquinolones. Cultures grown in sucrose peptone broth were dosed with enrofloxacin. After 21 days, 22% of the enrofloxacin remained. Three metabolites were identified: enrofloxacin N-oxide (62% of the total absorbance), N-acetylciprofloxacin (8.0%), and desethylene enrofloxacin (3.5%). PMID- 10831455 TI - trans-o-Hydroxybenzylidenepyruvate hydratase-aldolase as a biocatalyst. AB - The hydratase-aldolase-catalyzed conversion of trans-o-hydroxybenzylidenepyruvate to salicylaldehyde and pyruvate is an intermediate reaction in the conversion of naphthalene to salicylate by bacteria. Here, a variety of aromatic aldehydes and some nonaromatic aldehydes together with pyruvate have been shown to be substrates for aldol condensations catalyzed by this enzyme in extracts of the recombinant strain Escherichia coli JM109(pRE701). Some of the products of these reactions were also compared as substrates in the opposite (hydration-aldol cleavage) reaction. PMID- 10831456 TI - Detection of Pseudomonas savastanoi pv. savastanoi in olive plants by enrichment and PCR. AB - The sequence of the gene iaaL of Pseudomonas savastanoi EW2009 was used to design primers for PCR amplification. The iaaL-derived primers directed the amplification of a 454-bp fragment from genomic DNA isolated from 70 strains of P. savastanoi, whereas genomic DNA from 93 non-P. savastanoi isolates did not yield this amplified product. A previous bacterial enrichment in the semiselective liquid medium PVF-1 improved the PCR sensitivity level, allowing detection of 10 to 100 CFU/ml of plant extract. P. savastanoi was detected by the developed enrichment-PCR method in knots from different varieties of inoculated and naturally infected olive trees. Moreover, P. savastanoi was detected in symptomless stem tissues from naturally infected olive plants. This enrichment PCR method is more sensitive and less cumbersome than the conventional isolation methods for detection of P. savastanoi. PMID- 10831457 TI - Quantification of siderophore and hemolysin from Stachybotrys chartarum strains, including a strain isolated from the lung of a child with pulmonary hemorrhage and hemosiderosis. AB - A strain of Stachybotrys chartarum was recently isolated from the lung of a pulmonary hemorrhage and hemosiderosis (PH) patient in Texas (designated the Houston strain). This is the first time that S. chartarum has been isolated from the lung of a PH patient. In this study, the Houston strain and 10 strains of S. chartarum isolated from case (n = 5) or control (n = 5) homes in Cleveland were analyzed for hemolytic activity, siderophore production, and relatedness as measured by random amplified polymorphic DNA analysis. PMID- 10831458 TI - Fermentation of fructooligosaccharides by lactic acid bacteria and bifidobacteria. AB - Lactic acid bacteria and bifidobacteria were screened of their ability to ferment fructooligosaccharides (FOS) on MRS agar. Of 28 strains of lactic acid bacteria and bifidobacteria examined, 12 of 16 Lactobacillus strains and 7 of 8 Bifidobacterium strains fermented FOS. Only strains that gave a positive reaction by the agar method reached high cell densities in broth containing FOS. PMID- 10831459 TI - Isolation of a pandemic O3:K6 clone of a Vibrio parahaemolyticus strain from environmental and clinical sources in Thailand. AB - Application of an immunomagnetic enrichment method selective for Vibrio parahaemolyticus serovar K6 allowed isolation of a strain belonging to the pandemic O3:K6 clone of V. parahaemolyticus from fresh shellfish not implicated in a clinical case in southern Thailand. Arbitrarily primed PCR profiles of this strain, clinical O3:K6 strains isolated from sporadic diarrhea cases in the same area, and a standard pandemic O3:K6 strain were indistinguishable. PMID- 10831460 TI - Detection of human and animal rotavirus sequences in drinking water. AB - Reverse transcription-PCR analysis of drinking water in the homes of 56 children suffering from rotaviral gastroenteritis has shown the presence of the rotavirus genome in four samples. These strains were different from human rotaviruses detected in the children's feces, as determined by sequencing of the VP7 amplified fragments-three of them of animal origin (porcine or bovine) and one of human origin. PMID- 10831461 TI - Weighty perspectives: contemporary challenges in obesity and eating disorders. PMID- 10831462 TI - Obesity: what mental health professionals need to know. AB - OBJECTIVE: Obesity is a highly prevalent condition with significant health implications. This report summarizes recent clinically relevant findings concerning the pathogenesis and treatment of obesity and considers their implications for psychiatric diagnosis and management. METHOD: The authors conducted selective reviews of the literature from the last 10 years. Topics included the biological and behavioral factors that contribute to the onset and maintenance of obesity, the relationship between obesity and psychiatric illness and treatment, and the questions of whether and how obesity should be treated. RESULTS: Genetic effects, some mediated by eating behavior, contribute importantly to the potential for obesity, the expression of which is promoted by environmental factors that increase the availability of calorically dense foods and discourage activity. There appear to be behaviorally distinct subsets of obese persons who display particular patterns of disordered eating and elevated rates of psychopathology. Treatment with psychotropic medications may contribute to obesity in ways that are only partly understood. Although successful obesity treatment is associated with clear health benefits and available treatments offer benefit to some, relapse remains the rule. CONCLUSIONS: Although the presence or development of obesity is a daunting problem, it should not be ignored by mental health professionals. Treatment should address not only obesity per se, but also its effects on self-esteem in a hostile cultural climate. Ongoing developments in basic and clinical research are likely to increase the range, efficacy, and acceptability of treatment options in the years ahead. PMID- 10831463 TI - Neuropsychiatric symptoms associated with hepatitis C and interferon alpha: A review. AB - OBJECTIVE: Neuropsychiatric symptoms are commonly associated with chronic hepatitis C virus infection, its sequelae, and its treatment. In particular, interferon, a primary component of treatment for chronic hepatitis C, has been strongly associated with depressive symptoms. This review summarizes current knowledge about the etiology, course, and treatment of neuropsychiatric problems associated with hepatitis C and interferon alpha (IFN-alpha) treatment. METHOD: Studies were identified by computerized searches, and further references were obtained from bibliographies of the reviewed articles. RESULTS: Chronic infection with the hepatitis C virus is a common and growing problem, often affecting persons with psychiatric and substance use problems. Although changes in cognition, mood, and personality have been described in association with hepatitis C and with IFN-alpha treatment, there has been little systematic study of these changes. CONCLUSIONS: Psychiatrists should become familiar with the clinical spectrum associated with hepatitis C virus infection as well as the neuropsychiatric symptoms related to hepatitis C and IFN-alpha treatment. More studies are necessary to define the neuropsychiatric syndromes associated with this population and to find possible effective treatments. Furthermore, research is needed so that patients with psychiatric problems are not excluded from effective treatments for this growing medical problem. PMID- 10831464 TI - Images in neuroscience. Neural networks: neural systems I. PMID- 10831465 TI - First-episode psychosis in a managed care setting: clinical management and research. PMID- 10831467 TI - An empirical study of the classification of eating disorders. AB - OBJECTIVE: The nosology for eating disorders, despite having been extensively revised over time, may not capture the natural clustering of eating-related pathology as it occurs in general population samples. METHOD: Detailed information about anorectic and bulimic behaviors was assessed through personal interviews of 2,163 Caucasian female twins from a population-based registry. Latent class analysis was applied to nine eating disorder symptoms to develop an empirically based typology. Demographic, comorbidity, personality, and co-twin diagnosis data were used to validate the resultant classes. RESULTS: A six-class solution provided the best fit. One class displayed distorted eating attitudes without low body weight. Two classes demonstrated low weight without the psychological features of eating disorders. Three classes broadly resembled the DSM-IV classifications of anorexia nervosa, bulimia nervosa, and binge-eating disorder. For all classes, and especially for the three that reflected current diagnoses of eating disorders, monozygotic twins resembled one another much more in terms of class membership than did dizygotic twins. CONCLUSIONS: The authors found within a community sample, and through an empirical method, classes of eating-related pathology that broadly resembled the current classifications of anorexia nervosa, bulimia nervosa, and binge-eating disorder. Additional classes were marked by either the psychological features of eating disorders or low body weight. Individuals in the three eating-disorder classes had similar personality profiles but displayed differences in symptom expression and co-twin risk for anorexia nervosa, bulimia nervosa, and obesity. PMID- 10831468 TI - Adverse life events and cognitive-personality characteristics in the prediction of major depression and antidepressant response. AB - OBJECTIVE: Stressful life events are known to precipitate major depression. However, it remains unclear why some individuals who experience adverse events develop depression whereas others do not, and how the occurrence of life events affects treatment outcome. Emerging models posit that the effect of adverse life events varies by cognitive-personality style. This study examines the direct and interactive effects of stressful life events and cognitive-personality style in predicting 1) episode onset in patients with DSM-IV unipolar depression versus community comparison subjects and 2) depressive symptom severity at the completion of a 6-week standard antidepressant regimen. METHOD: Multivariate models were used to test the effects of adverse life events, cognitive personality style, and the congruence of event type (interpersonal versus achievement) with cognitive-personality style on depressive onset and treatment outcome in 43 patients with major depression and 43 healthy comparison subjects. Cognitive-personality characteristics were assessed by using Beck's measures of sociotropy (interpersonal dependency) and autonomy (need for independence and control). RESULTS: Adverse life events, sociotropy, and an autonomy factor need for control were each significantly related to depressive onset and predicted group status for 88% of the subjects. Event types affected outcome differently, and specific life event types interacted with cognitive-personality styles in predicting response to treatment. A multivariate model accounted for 65% of the variance in predicting outcome. CONCLUSIONS: Adverse life events are a potent factor in predicting depression. However, cognitive-personality characteristics also confer susceptibility to depression. Better outcome is associated with occurrence of adverse interpersonal events (e.g., death of a loved one) rather than adverse achievement events (e.g., loss of job) and occurs when the event type is congruent with cognitive-personality style. PMID- 10831469 TI - Transmission of response to trauma? Second-generation Holocaust survivors' reaction to cancer. AB - OBJECTIVE: Opinions differ about the effects of the Holocaust on the adult offspring of survivors. The authors studied cancer patients who were second generation Holocaust survivors in an attempt to determine whether they react to their illness with the high distress found to be a characteristic of Holocaust survivors. METHOD: In a study population of women with breast cancer, 106 second generation Holocaust survivors were compared to 102 women whose parents were not in the Holocaust. Background information was obtained by interviews. In addition, all patients completed three self-reports: the Mental Attitude to Cancer Scale, the Brief Symptom Inventory, and the Impact of Event Scale. RESULTS: The two groups had identical mean scores on the Mental Attitude to Cancer Scale. The offspring of Holocaust survivors had scores on the Brief Symptom Inventory and the Impact of Event Scale that were substantially and significantly higher and in the range of psychopathology. Within the group, married women and women whose mothers were still alive were even more distressed. Other independent variables shed little light on why the second-generation Holocaust survivors suffered from extreme distress. CONCLUSIONS: Second-generation Holocaust survivors are particularly vulnerable to psychological distress and, when faced with a trauma such as breast cancer, react with extreme psychological distress. PMID- 10831470 TI - Prospective study of posttraumatic stress, anxiety, and depressive reactions after earthquake and political violence. AB - OBJECTIVE: The authors sought to assess the severity and longitudinal course of posttraumatic stress, anxiety, and depressive reactions among two groups of adults differentially exposed to severe and mild earthquake trauma and a third group exposed to severe violence. They also examined interrelationships among these reactions and predictors of outcome and compared posttraumatic stress disorder (PTSD) symptom category profile and course between those exposed to earthquake and those exposed to violence. METHOD: Seventy-eight non-treatment seeking subjects were assessed with self-report instruments approximately 1.5 and 4.5 years after the 1988 Spitak earthquake in Armenia and the 1988 pogroms against Armenians in Azerbaijan. RESULTS: The two groups that had been exposed to severe trauma (earthquake or violence) had high initial and follow-up PTSD scores that did not remit over the 3-year interval. Overall, depressive symptoms subsided. Posttraumatic stress, anxiety, and depressive reactions were highly intercorrelated within and across both time intervals. No significant differences in PTSD severity, profile, or course were seen between subjects exposed to severe earthquake trauma versus those exposed to severe violence. CONCLUSIONS: After exposure to severe trauma, either an earthquake or violence, adults are at high risk of developing severe and chronic posttraumatic stress reactions that are associated with chronic anxiety and depressive reactions. Clinical evaluation and therapeutic intervention should include specific attention to these reactions. Early mental health intervention is recommended to prevent their chronicity. PMID- 10831471 TI - Minor physical anomalies and family adversity as risk factors for violent delinquency in adolescence. AB - OBJECTIVE: Minor physical anomalies are considered indicators of disruption in fetal development. They have been found to predict behavioral problems and psychiatric disorders. This study examined the extent to which minor physical anomalies, family adversity, and their interaction predict violent and nonviolent delinquency in adolescence. METHOD: Minor physical anomalies were assessed in a group of 170 adolescent boys from low socioeconomic status neighborhoods of Montreal. The boys had been enrolled in a longitudinal study since their kindergarten year, when an assessment of family adversity had been made on the basis of familial status and the parents' occupational prestige, age at the birth of the first child, and educational level. Adolescent delinquency was measured by using self-reported questionnaires and a search of official crime records. RESULTS: Results from logistic regression analyses indicated that both the total count of minor physical anomalies and the total count of minor physical anomalies of the mouth were significantly associated with an increased risk of violent delinquency in adolescence, beyond the effects of childhood physical aggression and family adversity. Similar findings were not found for nonviolent delinquency. CONCLUSIONS: Children with a higher count of minor physical anomalies, and especially a higher count of anomalies of the mouth, could be more difficult to socialize for different and additive reasons: they may have neurological deficits, and they may have feeding problems in the first months after birth. Longitudinal studies of infants with minor physical anomalies of the mouth are needed to understand the process by which they fail to learn to inhibit physical aggression. PMID- 10831472 TI - Effects of gonadal steroids in women with a history of postpartum depression. AB - OBJECTIVE: Endocrine factors are purported to play a role in the etiology of postpartum depression, but direct evidence for this role is lacking. The authors investigated the possible role of changes in gonadal steroid levels in postpartum depression by simulating two hormonal conditions related to pregnancy and parturition in euthymic women with and without a history of postpartum depression. METHOD: The supraphysiologic gonadal steroid levels of pregnancy and withdrawal from these high levels to a hypogonadal state were simulated by inducing hypogonadism in euthymic women-eight with and eight without a history of postpartum depression-with the gonadotropin-releasing hormone agonist leuprolide acetate, adding back supraphysiologic doses of estradiol and progesterone for 8 weeks, and then withdrawing both steroids under double-blind conditions. Outcome measures were daily symptom self-ratings and standardized subjective and objective cross-sectional mood rating scales. RESULTS: Five of the eight women with a history of postpartum depression (62.5%) and none of the eight women in the comparison group developed significant mood symptoms during the withdrawal period. Analysis of variance with repeated measures of daily and cross-sectional ratings of mood showed significant phase-by-group effects. These effects reflected significant increases in depressive symptoms in women with a history of postpartum depression but not in the comparison group after hormone withdrawal (and during the end of the hormone replacement phase), compared with baseline. CONCLUSIONS: The data provide direct evidence in support of the involvement of the reproductive hormones estrogen and progesterone in the development of postpartum depression in a subgroup of women. Further, they suggest that women with a history of postpartum depression are differentially sensitive to mood destabilizing effects of gonadal steroids. PMID- 10831473 TI - Five-year course and outcome of dysthymic disorder: A prospective, naturalistic follow-up study. AB - OBJECTIVE: There have been few naturalistic follow-up studies of dysthymic disorder. This study describes the 5-year course and outcome of dysthymic disorder. METHOD: The authors conducted a prospective, longitudinal follow-up study of 86 outpatients with early-onset dysthymic disorder and 39 outpatients with episodic major depressive disorder. Follow-ups, conducted 30 and 60 months after entry into the study, rated patients on the Longitudinal Interval Follow-Up Evaluation and the Modified Hamilton Rating Scale for Depression. RESULTS: The estimated 5-year recovery rate from dysthymic disorder was 52.9%. Among patients who recovered, the estimated risk of relapse was 45.2% during a mean of 23 months of observation. Patients with dysthymic disorder spent approximately 70% of the follow-up period meeting the full criteria for a mood disorder. During the course of the follow-up the patients with dysthymic disorder exhibited significantly greater levels of symptoms and lower functioning and were significantly more likely to attempt suicide and to be hospitalized than were patients with episodic major depressive disorder. Finally, among patients with dysthymic disorder who had never experienced a major depressive episode before entry into the study, the estimated risk of having a first lifetime major depressive episode was 76.9%. CONCLUSIONS: Dysthymic disorder is a chronic condition with a protracted course and a high risk of relapse. In addition, almost all patients with dysthymic disorder eventually develop superimposed major depressive episodes. Although patients with dysthymic disorder tend to show mild to moderate symptoms, from a longitudinal perspective, the condition is severe. PMID- 10831474 TI - Lost human capital from early-onset chronic depression. AB - OBJECTIVE: Chronic depression starts at an early age for many individuals and could affect their accumulation of "human capital" (i.e., education, higher amounts of which can broaden occupational choice and increase earnings potential). The authors examined the impact, by gender, of early- (before age 22) versus late-onset major depressive disorder on educational attainment. They also determined whether the efficacy and sustainability of antidepressant treatments and psychosocial outcomes vary by age at onset and quantified the impact of early versus late-onset, as well as never-occurring, major depressive disorder on expected lifetime earnings. METHOD: The authors used logistic and multivariate regression methods to analyze data from a three-phase, multicenter, double-blind, randomized trial that compared sertraline and imipramine treatment of 531 patients with chronic depression aged 30 years and older. These data were integrated with U.S. Census Bureau data on 1995 earnings by age, educational attainment, and gender. RESULTS: Early-onset major depressive disorder adversely affected the educational attainment of women but not of men. No significant difference in treatment responsiveness by age at onset was observed after 12 weeks of acute treatment or, for subjects rated as having responded, after 76 weeks of maintenance treatment. A randomly selected 21-year-old woman with early onset major depressive disorder in 1995 could expect future annual earnings that were 12%-18% lower than those of a randomly selected 21-year-old woman whose onset of major depressive disorder occurred after age 21 or not at all. CONCLUSIONS: Early-onset major depressive disorder causes substantial human capital loss, particularly for women. Detection and effective treatment of early onset major depressive disorder may have substantial economic benefits. PMID- 10831475 TI - Association between bipolar disorder and monoamine oxidase A gene polymorphisms: results of a multicenter study. AB - OBJECTIVE: Although genetic factors have been implicated in the etiology of bipolar disorder, no specific gene has been conclusively identified. Given the link between abnormalities in serotonergic neurotransmission and bipolar disorder, a candidate gene association approach was applied to study the involvement of the monoamine oxidase A (MAOA) gene, which codes for a catabolic enzyme of serotonin, in the susceptibility to bipolar disorder. METHOD: In France and Switzerland, 272 patients with bipolar disorder and 122 healthy subjects were typed for three polymorphic markers of the MAOA gene: the MAOA-CA repeat, the MAOA restriction fragment length polymorphism (RFLP), and a repeat directly adjacent to the variable number of tandem repeats (VNTR) locus. RESULTS: A significant difference in the distribution of the alleles for the MAOA-CA repeat was observed between the female bipolar patients and comparison group. CONCLUSIONS: The results obtained in the French and Swiss population confirm findings from two studies conducted in the United Kingdom. PMID- 10831476 TI - Anxiety as a correlate of response to the acute treatment of bipolar I disorder. AB - OBJECTIVE: Given the adverse impact of anxiety on treatment outcome in unipolar depression and the paucity of data on the role of anxiety in bipolar disorder, the authors sought to determine the effect of anxiety on the acute treatment response of patients with bipolar I disorder. METHOD: The authors examined the correlates of response to the acute treatment of 124 consecutively treated patients with bipolar I disorder. Measures of anxiety included history of panic attacks and a composite variable reflecting current or past anxiety symptoms. RESULTS: History of panic attacks proved to be a significant correlate of nonremission. Anxiety, as assessed with the composite variable, was associated with longer time to remission, as was the treatment of depressive versus manic symptoms and mixed versus manic symptoms. Patients with anxiety as assessed with the composite variable and patients with a history of panic attacks reported more severe medication side effects. They also required a greater number of medications, either sequentially or in combination, in order to achieve remission. CONCLUSIONS: The findings suggest that anxiety is a clinically meaningful correlate of poor outcome in the acute treatment of bipolar I disorder. PMID- 10831477 TI - ECT stimulus intensity: are present ECT devices too limited? AB - OBJECTIVE: The maximum output charge for ECT devices is limited to 576 millicoulombs in the United States, although there are no data ensuring that this limit will allow consistently effective treatments. The authors examined whether this limit has a negative impact on therapeutic response and, therefore, whether a higher stimulus charge should be available. METHOD: They retrospectively reviewed the records of 471 patients who received a clinical index course of ECT at Duke University between 1991 and 1998. These patients received conservative stimulus dosing of 2.25 times seizure threshold for unilateral ECT and 1.5 times seizure threshold for bilateral ECT. RESULTS: Seventy-two (15%) of the 471 patients required the maximum stimulus intensity during their index ECT course. Of these, 24 (5% of the total) had either a short EEG seizure (less than 25 seconds) or had no seizure at the maximum level. Strategies to augment therapeutic response with caffeine, ketamine, or hyperventilation were used in 14 of the 24 patients, and data on therapeutic response were available for 22 of the 24. Only seven (32%) of these 22 patients were considered ECT responders, compared with 242 (66%) of the remaining 364 patients for whom data on response to ECT were available. Older age and pre-ECT course EEG slowing were predictors of requiring the maximum stimulus level. CONCLUSIONS: The maximum available stimulus output was therapeutically insufficient for 5% of the patients studied even when available means to augment response were instituted. This percentage would likely be even larger with the use of a less conservative dosing protocol for unilateral ECT. Increases in maximum stimulus output for ECT devices should be considered as a means to ensure adequate treatment response. PMID- 10831478 TI - Efficacy of extended-release venlafaxine in nondepressed outpatients with generalized anxiety disorder. AB - OBJECTIVE: This study evaluated the efficacy and safety of fixed doses of once daily extended-release (XR) venlafaxine in outpatients with generalized anxiety disorder without concomitant major depressive disorder. METHOD: Adult outpatients with generalized anxiety disorder but not major depressive disorder with total scores of 18 or higher on the Hamilton Rating Scale for Anxiety and scores of 2 or higher on its anxious mood and tension factors were eligible. Patients were randomly assigned to receive placebo or venlafaxine XR (75, 150, or 225 mg/day) for 8 weeks. Primary efficacy variables were final total and psychic anxiety factor scores on the Hamilton anxiety scale and final severity and global improvement item scores on the Clinical Global Impression (CGI) scale. RESULTS: Of the 377 patients entering the study, 370 were included in a safety analysis and 349 in an efficacy analysis. Adjusted mean scores at 8 weeks (last observation-carried-forward analysis) were significantly lower for one or more of the venlafaxine XR groups in four of four primary and three of four secondary outcome measures than for the placebo group. These included a change of 1.7 (versus 1.3) from baseline on CGI severity item scores and a final score of 2.2 (versus 2.6) on the CGI global improvement item. All doses of venlafaxine XR were well tolerated. CONCLUSIONS: Venlafaxine XR is an effective and well-tolerated option for the short-term treatment of generalized anxiety disorder in outpatients without major depressive disorder. PMID- 10831479 TI - Clozapine, diabetes mellitus, weight gain, and lipid abnormalities: A five-year naturalistic study. AB - OBJECTIVE: The goal of this 5-year naturalistic study of patients treated with clozapine was to examine the incidence of treatment-emergent diabetes mellitus in relation to other factors, including weight gain, lipid abnormalities, age, clozapine dose, and treatment with valproate. METHOD: Data on age, gender, race, diagnosis, family history of diabetes, and age at clozapine initiation were collected from medical records of 82 outpatients with schizophrenia or schizoaffective disorder. Clozapine dose, data on use of valproate, and laboratory test results were recorded at 6-month intervals. RESULTS: The mean age at the time of clozapine initiation of the 82 patients was 36.4 years; 26.8% of the patients were women, and 91.5% were Caucasian. The mean baseline weight was 175.5 lb, and the mean body mass index was 26.9 kg/m(2). Thirty patients (36.6%) were diagnosed with diabetes during the 5-year follow-up. Weight gain, use of valproate, and total daily dose of clozapine were not significant risk factors for developing diabetes mellitus. Patients experienced significant weight gain that continued until approximately month 46 from initiation of clozapine. There was a nonsignificant increase in total serum cholesterol and a significant increase in serum triglycerides level. CONCLUSIONS: The results support the hypotheses that patients treated with clozapine experience significant weight gain and lipid abnormalities and appear to be at increased risk for developing diabetes. PMID- 10831480 TI - Clozapine in the treatment of refractory psychotic mania. AB - OBJECTIVE: The efficacy of clozapine was examined in a group of patients with treatment-refractory bipolar disorder, manic type with psychotic features. METHOD: Twenty-two subjects with treatment-refractory bipolar disorder with active manic and psychotic symptoms participated in a 12-week open-label trial of clozapine. After a 2-10-day drug washout period, patients began treatment with clozapine at 25 mg/day; the dose was increased 25 mg/day (as tolerated) to a maximum level of 550 mg/day. Patients were evaluated longitudinally over the course of the study with the Brief Psychiatric Rating Scale (BPRS), the Young Mania Rating Scale, and the Clinical Global Impressions (CGI) scale. RESULTS: Fourteen of the 22 subjects in the study continued taking clozapine for at least 10 of the 12 weeks. Among the entire group, mean improvements of 56. 7%, 56.6%, and 39.1% were seen on the BPRS, Young Mania Rating Scale, and CGI, respectively. Seventeen of the 22 subjects (77.3%) experienced at least a 20% improvement in scores on all three scales. CONCLUSIONS: The findings from this open-label study, which are consistent with previous retrospective studies, case reports, and one other open-label prospective study, suggest that clozapine is an effective agent for patients with treatment-refractory psychotic mania. PMID- 10831481 TI - Differential efficacy of olanzapine for deficit and nondeficit negative symptoms in schizophrenia. AB - OBJECTIVE: Atypical antipsychotic medications have generally been found to be more effective than conventional antipsychotics in the treatment of negative symptoms. Whether the benefits derived from the atypical agents are the result of improvements in primary versus secondary negative symptoms is unclear. The authors examined the effects of olanzapine on primary and secondary negative symptoms for patients with severe negative symptoms who did or did not have the deficit syndrome. METHOD: Thirty-nine outpatients with schizophrenia and severe negative symptoms were assessed for the presence of the deficit syndrome and entered into a 12-week, open-label study of olanzapine. Positive and negative symptoms, extrapyramidal side effects, quality of life, and level of functioning of the patients were assessed at baseline and endpoint. RESULTS: All 39 patients completed the 12-week protocol; 13 of the patients had deficit negative symptoms, and 26 had nondeficit negative symptoms. Patients who had nondeficit negative symptoms demonstrated improvements in positive and negative symptoms, level of functioning, and extrapyramidal side effects over baseline. In contrast, patients meeting criteria for the deficit syndrome improved significantly over baseline only in extrapyramidal side effects. CONCLUSIONS: The results of this study suggest that olanzapine is efficacious for secondary negative symptoms in schizophrenia but fail to support the contention that olanzapine has a direct beneficial effect on primary negative symptoms. PMID- 10831482 TI - Relation of familial schizophrenia to negative symptoms but not to the deficit syndrome. AB - OBJECTIVE: Although a family history of schizophrenia has been associated with negative symptoms, family history is inconsistently related to the presence of the deficit syndrome. METHOD: The authors assessed family history and the deficit syndrome in 99 patients with DSM-III-R-diagnosed schizophrenia who were assessed during clinical treatment. Of these 99 patients, 45 were assessed both while antipsychotic free and during antipsychotic treatment to index their treatment response. RESULTS: Patients with (N=39) and without (N=60) a family history of schizophrenia had similar proportions of the deficit syndrome. Yet family history and deficit syndrome categorizations identified a group with greater negative symptoms on the Positive and Negative Syndrome Scale. Those with a family history had greater emotional withdrawal, poor rapport, and lack of spontaneity. Groups with and without the deficit syndrome similarly differed in these symptoms but also in affective blunting, motor retardation, and passive or apathetic social withdrawal. The study involving antipsychotic-free and antipsychotic treatment phases showed main medication effects explaining positive, psychopathology, depression, and activation symptoms but not negative symptoms. Only patients without a family history had improved negative symptoms with antipsychotic treatment. CONCLUSIONS: Patients with a family history of schizophrenia had greater and more treatment-resistant negative symptoms than those without a family history. They were not more likely to have the deficit syndrome. The group with a family history had more pathology only in negative symptoms related to psychosocial function. The stable negative symptoms specifically related to the genetic vulnerability to inherit schizophrenia might be those associated with psychosocial functioning. PMID- 10831483 TI - Placebo-controlled trial of sertraline in the treatment of binge eating disorder. AB - OBJECTIVE: The authors' goal was to assess the efficacy of sertraline in the treatment of binge eating disorder. METHOD: Thirty-four outpatients with DSM-IV binge eating disorder were randomly assigned to receive either sertraline (N=18) or placebo (N=16) in a 6-week, double-blind, flexible-dose (50-200 mg) study. Except for response level, outcome measures were analyzed by random regression methods, with treatment-by-time interaction as the effect measure. RESULTS: Compared with placebo, sertraline was associated with a significantly greater rate of reduction in the frequency of binges, clinical global severity, and body mass index as well as a significantly greater rate of increase in clinical global improvement. Patients receiving sertraline who completed the study demonstrated a higher level of response, although the effect was not significant. CONCLUSIONS: In a 6-week trial, sertraline was effective and well tolerated in the treatment of binge eating disorder. PMID- 10831484 TI - Platelet activation in depression and effects of sertraline treatment: An open label study. AB - OBJECTIVE: The authors' goal was to determine whether sertraline attenuates the increased platelet activation seen among depressed patients. METHOD: They tested 21 otherwise healthy patients with untreated major depressive episode who were 25 52 years old and 21 age- and sex-matched comparison subjects. Patients received 6 weeks of sertraline treatment, and 17 returned for retesting. RESULTS: At baseline, the depressed patients had greater platelet secretion than the comparison subjects in response to collagen. Depressed patients with a family history of coronary disease had nonsignificantly greater wound-induced fibrinogen receptor binding than the other subjects. Platelet secretion in response to collagen was significantly reduced after treatment with sertraline. CONCLUSIONS: Sertraline diminished the increased platelet secretion found among depressed patients, although the findings are limited by a lack of a placebo control group. PMID- 10831485 TI - Effect of cognitive impairment and premorbid intelligence on treatment preferences for life-sustaining medical therapy. AB - OBJECTIVE: This study examines the influence of cognitive impairment, premorbid intelligence, and decision-making capacity to complete advance directives on the treatment preferences for life-sustaining medical therapy in the elderly. METHOD: One hundred elderly individuals were recruited. Fifty were first referrals to specialist services with a DSM-IV diagnosis of dementia, and 50 were volunteers. Each person was asked about treatment preferences in three clinical vignettes. RESULTS: Elderly individuals who had cognitive impairment and were incapable of completing advance directives were significantly more likely to opt for life sustaining interventions. There was no association between premorbid intelligence and treatment preferences. CONCLUSIONS: Cognitive impairment appears to influence treatment preferences for life-sustaining medical therapy. With increasing cognitive impairment, elderly individuals tend to opt for treatment interventions. PMID- 10831486 TI - Dissociative disorders in Dutch psychiatric inpatients. AB - OBJECTIVE: The goal of this study was to determine the frequency of dissociative disorders in Dutch psychiatric inpatients. METHOD: During a period of 12 months, 122 consecutively admitted adult psychiatric patients were screened with the Dissociative Experiences Scale. Patients scoring 25 and higher and a random selection of patients scoring lower than 25 were blindly interviewed with the Structured Clinical Interview for DSM-IV Dissociative Disorders, Revised. Interviews were scored independently by a blind rater. RESULTS: Ten (8%) of the 122 patients were diagnosed as having a dissociative disorder; two (2%) were diagnosed as having a dissociative identity disorder. Two patients (2%) had factitious dissociative identity disorder. CONCLUSIONS: The rate of dissociative disorders in this group of Dutch patients is comparable to the rates reported in other European studies but lower than rates reported in North American studies. PMID- 10831487 TI - Is placebo response the same as drug response in panic disorder? AB - OBJECTIVE: The authors used seven definitions of response in panic disorder to compare patient-rated improvements in quality of life between patients with panic disorder who responded to sertraline and those who responded to placebo. METHOD: They combined and examined data from two multicenter, randomized, double-blind, parallel-group, flexible-dose studies of panic disorder (N=302). RESULTS: Significant differences in quality of life between patients who responded to sertraline and those who responded to placebo were apparent across all the definitions of clinical response. CONCLUSIONS: Patients who respond to placebo in panic disorder treatment studies may show symptom relief but may not experience improvement in quality of life. Determinations of quality of life should be included as components of both standard clinical assessment and clinical treatment studies of patients with panic disorder. PMID- 10831488 TI - Decreased serotonin 2A receptor densities in neuroleptic-naive patients with schizophrenia: A PET study using [(18)F]setoperone. AB - OBJECTIVE: The authors compared serotonin receptor binding in patients with schizophrenia and healthy comparison subjects. METHOD: They used positron emission tomography with [(18)F]setoperone to examine six patients with schizophrenia who had never been given neuroleptics and seven age-matched subjects who did not have schizophrenia. RESULTS: A nondirected voxel-based analysis of the subjects' entire search volume found that serotonin 2A binding potential in the frontal cortex index was significantly smaller (by 16.3%) in patients with schizophrenia than in healthy subjects. CONCLUSIONS: The authors conclude that the decrease in serotonin receptor densities previously reported in postmortem studies of subjects with schizophrenia are present at the onset of the illness, before exposure to neuroleptics. PMID- 10831489 TI - Subjective experience and striatal dopamine D(2) receptor occupancy in patients with schizophrenia stabilized by olanzapine or risperidone. AB - OBJECTIVE: The authors' goal was to study the relationship between subjective experience during treatment with olanzapine or risperidone and dopamine D(2) receptor occupancy in stabilized patients with schizophrenia. METHOD: Subjective experience, psychopathology, and extrapyramidal symptoms were assessed, and D(2) receptor occupancy was determined with [(123)I]iodobenzamide single photon emission computed tomography, in 22 patients whose schizophrenia was stabilized by olanzapine or risperidone. RESULTS: Subjective experience, depression, and negative symptoms were related to dopamine D(2) receptor occupancy, but extrapyramidal symptoms were not. CONCLUSIONS: These results provide preliminary evidence that negative subjective experience is related to high D(2) receptor occupancy. Longitudinal study is required because this relationship may have implications for dosing strategies. PMID- 10831490 TI - Exacerbation of psychosis by phenylpropanolamine. PMID- 10831491 TI - Retreatment with clozapine after erythromycin-induced neutropenia. PMID- 10831492 TI - Acute onset of auditory hallucinations after initiation of celecoxib therapy. PMID- 10831493 TI - Serotonin syndrome from addition of low-dose trazodone to nefazodone. PMID- 10831494 TI - Parenteral valproate for control of acute mania. PMID- 10831495 TI - Schizophrenias in the Wernicke-Kleist-Leonhard school. PMID- 10831496 TI - Weight criteria for diagnosis of anorexia nervosa. PMID- 10831498 TI - Medication treatment versus cognitive behavior therapy. PMID- 10831500 TI - Short screening scale for posttraumatic stress disorder. PMID- 10831514 TI - May 30, 2000 PMID- 10831515 TI - Relation between endothelial cell apoptosis and blood flow direction in human atherosclerotic plaques. AB - BACKGROUND: Blood flow characteristics influence endothelial cell apoptosis. However, little is known about the occurrence of endothelial cell apoptosis in human atherosclerosis and its relation to blood flow. METHODS AND RESULTS: A total of 42 human carotid atherosclerotic plaques were retrieved by endarterectomy; they were examined in the longitudinal axial direction. Plaques were included in this study when upstream and downstream parts were clearly visible, occlusion was absent, and immunostaining for luminal endothelium was present all along the plaque. Using these criteria, 13 plaques were processed for further immunohistochemical studies (using anti-CD31, anti-Ki-67, and anti splicing factor antibodies) and in situ detection of apoptosis (terminal dUTP nick end-labeling and ligase assay). Eight plaques showed > or =1 apoptotic endothelial cell at the luminal surface. Quantitative analysis of endothelial cell apoptosis in these plaques showed a systematic preferential occurrence of apoptosis in the downstream parts of plaques, where low flow and low shear stress prevail, in comparison with the upstream parts (18.8+/-3.3% versus 2.7+/-1.2%, respectively, P<0.001). Endothelial cell apoptosis was barely detectable in plaque microvessels. CONCLUSIONS: Our results suggest that in vivo local shear stress influences luminal endothelial cell apoptosis and may be a major determinant of plaque erosion and thrombosis. PMID- 10831516 TI - Edge restenosis after implantation of high activity (32)P radioactive beta emitting stents. AB - BACKGROUND: A high restenosis rate has been reported at the edges ("edge restenosis") of (32)P radioactive stents with an initial activity level of 3 to 12 microCi. This edge effect might be due to balloon injury and to a low dose of radiation at the stent margins. The aim of this study was to evaluate whether the implantation of (32)P radioactive stents with a higher activity level (12 to 21 microCi) combined with a nonaggressive stent implantation strategy could solve the problem of edge restenosis. METHODS AND RESULTS: We compared the results of lesions treated with single radioactive BX stents with an activity of 12 to 21 microCi (group 2, n = 54 lesions) with the results of lesions treated by single radioactive BX stents with an initial activity level of 3 to 12 microCi (group 1, n = 42 lesions). There were no procedural events. At the 6-month follow-up, no myocardial infarctions, deaths, or stent thromboses had occurred. Intrastent binary restenosis was 0% in group 1 versus 4% in group 2 (n = 2, both at the ostium of the right coronary artery, P = NS). Intrastent neointimal hyperplasia was significantly lower in group 2 than in group 1. The intralesion (intrastent plus peri-stent) restenosis rate was 38% in group 1 versus 30% in group 2 (P = NS). Conclusions-Single (32)P radioactive stents with an initial activity level of 12 to 21 microCi reduced intrastent neointimal hyperplasia compared with stents of 3 to 12 microCi, but they did not solve the problem of edge restenosis, even if a nonaggressive stent implantation strategy was used. PMID- 10831517 TI - Left ventricular apex ablation decreases the upper limit of vulnerability. AB - BACKGROUND: After shocks with an approximately 50% probability of success for the upper limit of vulnerability (ULV(50)) of strength, the first few activations appear focally on the epicardium at almost the same site at the left ventricular (LV) apex in both successful and failed induction of ventricular fibrillation (VF). We tested the hypothesis that subendocardial ablation at this early site would decrease the shock strength required for the ULV(50). METHODS AND RESULTS: Ten S1 stimuli were delivered from the right ventricular apex at a 300-ms coupling interval in 5 pigs. Biphasic shocks were delivered from right ventricular-superior vena cava electrodes after the last S1 stimulus. The ULV(50) was determined using an up/down protocol with T-wave scanning. Radiofrequency ablation was performed endocardially at the apical LV. The ULV(50) was determined again 30 minutes after ablation. To determine the importance of the ablation region, this protocol was repeated in another 5 pigs with ablation at the LV base. Delivered voltage (401+/-60 versus 323+/-50 V) and energy (11+/-3 versus 7+/-2 J) for the ULV(50) were significantly decreased after LV apex ablation by 19% and 34%, respectively. However, no difference existed in ULV(50) before and after LV base ablation. Lesions at both the LV apex and base were subendocardial and ranged from 0.8 to 1.1 cm in diameter. CONCLUSIONS: Subendocardial ablation at the apical LV markedly decreases ULV(50), which suggests that the activation originating from this postshock early site is responsible for VF initiation and that interventions to electrically silence this site can influence the outcome of VF induction by ULV shocks. PMID- 10831518 TI - Linkage of the cholesteryl ester transfer protein (CETP) gene to LDL particle size: use of a novel tetranucleotide repeat within the CETP promoter. AB - BACKGROUND: A preponderance of small, dense LDL particles, elevated levels of plasma triglycerides (TG), and low levels of HDL characterize the atherogenic lipoprotein phenotype, which is associated with increased coronary artery disease (CAD) risk. Genetic and environmental factors influence LDL size, cholesteryl ester transfer protein (CETP) being one of the candidate genes. CETP mediates the transfer of cholesteryl ester from HDL to apolipoprotein (apo) B-containing lipoproteins in exchange for TG, promoting reverse cholesterol transfer and remodeling of lipoprotein particles. METHODS AND RESULTS: We have identified a tetranucleotide repeat (fragment sizes from 324 to 464 bp; heterozygosity index = 0.74) within the CETP promoter and used it in quantitative sib-pair linkage analysis in 119 female dizygotic (DZ) twins. Linkage was found to LDL size (P<0.001), TG (P<0.005), and plasma apoB (P = 0.02). The distribution of the tetranucleotide repeats was bimodal, and there was strong allelic association of the "short" alleles with the B2 allele of CETP TaqIB polymorphic site (P<0.001). CONCLUSIONS: This report of linkage of the CETP gene to LDL particle size adds to the list of candidate genes linked to LDL size, supporting the hypothesis of multigenic determination of LDL size heterogeneity. Whether this promoter variation is itself functional or is a marker for a functional site in the CETP gene remains to be determined. PMID- 10831519 TI - Geographic miss: a cause of treatment failure in radio-oncology applied to intracoronary radiation therapy. AB - BACKGROUND: A recognized limitation of endovascular beta-radiation therapy is the development of new stenosis at the edges of the irradiated area. The combination of injury and low-dose radiation may be the precursor of this phenomenon. We translated the radio-oncological concept of "geographic miss" to define cases in which the radiation source did not fully cover the injured area. The aims of the study were to determine the incidence and causes of geographic miss and evaluate the impact of this inadequate treatment on the outcome of patients treated with intracoronary beta-radiation. METHODS AND RESULTS: We analyzed 50 consecutive patients treated with beta-radiation after percutaneous coronary intervention. The prescribed dose ranged between 12 and 20 Gy at 2 mm from the source axis. By means of quantitative coronary angiography, the irradiated segment (IRS) and both edges were studied before and after intervention and at 6-month follow-up. Edges that were injured during the procedure constituted the geographic miss edges. Twenty-two edges were injured during the intervention, mainly because of procedural complications that extended the treatment beyond the margins of the IRS. Late loss was significantly higher in geographic miss edges than in IRSs and uninjured edges (0.84+/-0.6 versus 0.15+/-0.4 and 0.09+/-0.4 mm, respectively; P<0.0001). Similarly, restenosis rate was significantly higher in the injured edges (10% within IRS, 40.9% in geographic miss edges, and 1.9% in uninjured edges; P<0.001). CONCLUSIONS: These data support the hypothesis that the combination of injury and low-dose beta-radiation induces deleterious outcome. PMID- 10831520 TI - Residual plaque burden, delivered dose, and tissue composition predict 6-month outcome after balloon angioplasty and beta-radiation therapy. AB - BACKGROUND: Inhomogeneity of dose distribution and anatomic aspects of the atherosclerotic plaque may influence the outcome of irradiated lesions after balloon angioplasty (BA). We evaluated the influence of delivered dose and morphological characteristics of coronary stenoses treated with beta-radiation after BA. METHODS AND RESULTS: Eighteen consecutive patients treated according to the Beta Energy Restenosis Trial 1.5 were included in the study. The site of angioplasty was irradiated with the use of a beta-emitting (90)Sr/(90)Y source. With the side branches used as anatomic landmarks, the irradiated area was identified and volumetric assessment was performed by 3D intracoronary ultrasound imaging after treatment and at 6 months. The type of tissue, the presence of dissection, and the vessel volumes were assessed every 2 mm within the irradiated area. The minimal dose absorbed by 90% of the adventitial volume (D(v90)Adv) was calculated in each 2-mm segment. Diffuse calcified subsegments and those containing side branches were excluded. Two hundred six coronary subsegments were studied. Of those, 55 were defined as soft, 129 as hard, and 22 as normal/intimal thickening. Plaque volume showed less increase in hard segments as compared with soft and normal/intimal thickening segments (P<0.0001). D(v90)Adv was associated with plaque volume at follow-up after a polynomial equation with linear and nonlinear components (r = 0.71; P = 0.0001). The multivariate regression analysis identified the independent predictors of the plaque volume at follow-up: plaque volume after treatment, D(v90)Adv, and type of plaque. CONCLUSIONS: Residual plaque burden, delivered dose, and tiss composition play a fundamental role in the volumetric outcome at 6-month follow-up after beta-radiation therapy and BA. PMID- 10831521 TI - Increased risk of restenosis after placement of gold-coated stents: results of a randomized trial comparing gold-coated with uncoated steel stents in patients with coronary artery disease. AB - BACKGROUND: Gold is a highly biocompatible material. Experimental evidence suggests that coating the stent with a gold layer may have a beneficial influence. In this randomized trial, we assessed whether gold-coated stents were associated with a better clinical and angiographic outcome after coronary placement. METHODS AND RESULTS: Patients with symptomatic coronary artery disease were randomly assigned to receive either a gold-coated Inflow stent (n = 367) or an uncoated Inflow stainless steel stent (n = 364) of identical design. Follow-up angiography was routinely performed at 6 months. The primary end point of the study was the occurrence of any adverse clinical event (death, myocardial infarction, or target-vessel revascularization) during the first year after stenting. At 30 days, there was no significant difference in the combined incidence of adverse events, with 7.9% in the gold-stent group versus 5.8% in the steel-stent group (P = 0.25). The incidence of angiographic restenosis (> or =50% diameter stenosis) was 49.7% in the gold-stent group and 38.1% in the steel-stent group (P = 0.003). One-year survival free of myocardial infarction was 88.6% in the gold-stent group and 91.8% in the steel-stent group (P = 0.14). One-year event-free survival was significantly less favorable in the gold-stent group (62.9% versus 73.9% in the steel-stent group; P = 0.001). CONCLUSIONS: Coating steel stents with gold had no significant influence on the thrombotic events observed during the first 30 days after the intervention. However, gold-coated stents were associated with a considerable increase in the risk of restenosis over the first year after stenting. PMID- 10831522 TI - Treatment of in-stent restenosis with excimer laser coronary angioplasty versus rotational atherectomy: comparative mechanisms and results. AB - BACKGROUND: Atheroablation yields improved clinical results for balloon angioplasty (percutaneous transluminal coronary angioplasty, PTCA) in the treatment of diffuse in-stent restenosis (ISR). METHODS AND RESULTS: We compared the mechanisms and clinical results of excimer laser coronary angioplasty (ELCA) versus rotational atherectomy (RA), both followed by adjunct PTCA; 119 patients (158 ISR lesions) were treated with ELCA+PTCA and 130 patients (161 ISR lesions) were treated with RA+PTCA. Quantitative coronary angiographic and planar intravascular ultrasound (IVUS) measurements were performed routinely. In addition, volumetric IVUS analysis to compare the mechanisms of lumen enlargement was performed in 28 patients with 30 lesions (16 ELCA+PTCA, 14 RA+PTCA). There were no significant between-group differences in preintervention or final postintervention quantitative coronary angiographic or planar IVUS measurements of luminal dimensions. Angiographic success and major in-hospital complications with the 2 techniques were also similar. Volumetric IVUS analysis showed significantly greater reduction in intimal hyperplasia volume after RA than after ELCA (43+/-14 versus 19+/-10 mm(3), P<0.001) because of a significantly higher ablation efficiency (90+/-10% versus 76+/-12%, P = 0.004). However, both interventional strategies had similar long-term clinical outcome; 1-year target lesion revascularization rate was 26% with ELCA+PTCA versus 28% with RA+PTCA (P = NS). CONCLUSIONS: Despite certain differences in the mechanisms of lumen enlargement, both ELCA+PTCA and RA+PTCA can be used to treat diffuse ISR with similar clinical results. PMID- 10831523 TI - Clinical profile and outcome of idiopathic restrictive cardiomyopathy. AB - BACKGROUND: Idiopathic restrictive cardiomyopathy is a poorly recognized entity of unknown cause characterized by nondilated, nonhypertrophied ventricles with diastolic dysfunction resulting in dilated atria and variable systolic function. METHODS AND RESULTS: Between 1979 and 1996, 94 patients (61% women) 10 to 90 years old (mean, 64 years) met strict morphological echocardiographic criteria for idiopathic restrictive cardiomyopathy, mainly dilated atria with nonhypertrophied, nondilated ventricles. None had known infiltrative disease, hypertension of >5 years' duration, or cardiac or systemic conditions associated with restrictive filling. Nineteen percent were in NYHA class I, 53% in class II, and 28% in class III or IV. Atrial fibrillation was noted in 74% of patients and systolic dysfunction in 16%. Follow-up (mean, 68 months) was complete for 93 patients (99%). At follow-up, 47 patients (50%) had died, 32 (68%) of cardiovascular causes. Four had heart transplantation. The death rate compared with actuarial statistics was significantly higher than expected (P<0.0001). Kaplan-Meier 5-year survival was 64%, compared with expected survival of 85%. Multivariate analysis using proportional hazards showed that the risk of death approximately doubles with male sex (hazard ratio [HR] = 2.1), left atrial dimension >60 mm (HR = 2.3), age >70 years (HR = 2.0), and each increment of NYHA class (HR = 2.0). CONCLUSIONS: Idiopathic restrictive cardiomyopathy or nondilated, nonhypertrophic ventricles with marked biatrial dilatation, as defined morphologically by echocardiography, affects predominantly elderly patients but can occur in any age group. Patients present with systemic and pulmonary venous congestion and atrial fibrillation and have a poor prognosis, particularly men >70 years old with higher NYHA class and left atrial dimension >60 mm. PMID- 10831524 TI - New insights into the progression of aortic stenosis: implications for secondary prevention. AB - BACKGROUND: The risk factors affecting aortic stenosis (AS) progression are not clearly defined. Insights into this may allow for its secondary prevention. METHODS AND RESULTS: We investigated predictors of AS progression in 170 consecutive patients with AS who had paired echocardiograms > or =3 months (23+/ 11) apart. Various clinical, echocardiographic, and biochemical variables were related to the change in aortic valve area (AVA). The annual rate of reduction in AVA was 0.10+/-0.27 cm(2) or 7+/-18% per year. The reduction in AVA per year was significantly related to initial AVA (r = 0.46, P<0.0001), the mean aortic valve gradient (r = 0.27, P = 0.04), left ventricular (LV) outflow tract velocity (r = 0.26, P = 0.001), and LV end-diastolic diameter (r = 0.20, P = 0.04) and marginally to serum creatinine level (r = 0.15, P = 0.08). Patients with a rate of reduction in AVA faster than the mean had higher serum creatinine (P = 0.04) and calcium (P = 0.08) levels. Those with a serum cholesterol level >200 mg/dL had a rate of AVA reduction roughly twice that of those with a lower cholesterol level (P = 0.04). Stepwise multiple regression analysis identified initial AVA, current smoking, and serum calcium level as the independent predictors of amount of AVA reduction per year. CONCLUSIONS: Absolute and percentage reduction in AVA per year in those with AS is greater in those with milder degrees of stenosis and is accelerated in the presence of smoking, hypercholesterolemia, and elevated serum creatinine and calcium levels. These findings may have important implications in gaining further insights into the mechanism of AS progression and in formulating strategies to retard this process. PMID- 10831525 TI - In vivo magnetic resonance evaluation of atherosclerotic plaques in the human thoracic aorta: a comparison with transesophageal echocardiography. AB - BACKGROUND: The structure and composition of aortic atherosclerotic plaques are associated with the risk of future cardiovascular events. Magnetic resonance (MR) imaging may allow accurate visualization and characterization of aortic plaques. METHODS AND RESULTS: We developed a noninvasive MR method, free of motion and blood flow artifacts, for submillimeter imaging of the thoracic aortic wall. MR imaging was performed on a clinical MR system in 10 patients with aortic plaques identified by transesophageal echocardiography (TEE). Plaque composition, extent, and size were assessed from T1-, proton density-, and T2- weighted images. Comparison of 25 matched MR and TEE cross-sectional aortic plaque images showed a strong correlation for plaque composition (chi(2) = 43.5, P<0.0001; 80% overall agreement; n = 25) and mean maximum plaque thickness (r = 0.88, n = 25; 4.56+/ 0.21 mm by MR and 4.62+/-0.31 mm by TEE). Overall aortic plaque extent as assessed by TEE and MR was also statistically significant (chi(2) = 61.77, P<0.0001; 80% overall agreement; n = 30 regions). CONCLUSIONS: This study demonstrates that noninvasive MR evaluation of the aorta compares well with TEE imaging for the assessment of atherosclerotic plaque thickness, extent, and composition. This MR method may prove useful for the in vivo study of aortic atherosclerosis. PMID- 10831526 TI - Human serum paraoxonases (PON1) Q and R selectively decrease lipid peroxides in human coronary and carotid atherosclerotic lesions: PON1 esterase and peroxidase like activities. AB - BACKGROUND: Human serum paraoxonase (PON1) exists in two polymorphic forms: one that differs in the amino acid at position 192 (glutamine and arginine, Q and R, respectively) and the second one that differs in the amino acid at position 55 (methionine and leucine, M and L, respectively). PON1 protects LDL from oxidation, and during LDL oxidation, PON1 is inactivated. METHODS AND RESULTS: The present study compared PON1 isoforms Q and R for their effect on lipid peroxide content in human coronary and carotid lesions. After 24 hours of incubation with PON1Q or PON1R (10 arylesterase units/mL), lipid peroxides content in both coronary and carotid lesion homogenates (0.1 g/mL) was reduced up to 27% and 16%, respectively. The above incubation was associated with inactivation of PON1Q and PON1R by 15% and 45%, respectively. Lesion cholesteryl linoleate hydroperoxides and cholesteryl linoleate hydroxides were hydrolyzed by PON1 to yield linoleic acid hydroperoxides and linoleic acid hydroxides. Furthermore, lesion and pure linoleic acid hydroperoxides were reduced to yield linoleic acid hydroxides. These results thus indicate that PON1 demonstrates esterase-like and peroxidase-like activities. Recombinant PON1 mutants in which the PON1-free sulfhydryl group at cysteine-284 was replaced with either alanine or serine were no longer able to reduce lipid peroxide content in carotid lesions. CONCLUSIONS: We conclude that PON1 may be antiatherogenic because it hydrolyzes lipid peroxides in human atherosclerotic lesions. PMID- 10831527 TI - Soluble tumor necrosis factor receptor abrogates myocardial inflammation but not hypertrophy in cytokine-induced cardiomyopathy. AB - BACKGROUND: Transgenic mice with cardiac-specific overexpression of tumor necrosis factor (TNF)-alpha develop dilated cardiomyopathy. The present study was designed to evaluate therapeutic effects of adenovirus-mediated neutralization of TNF-alpha on this model. METHODS AND RESULTS: An adenovirus encoding the 55-kDa TNF receptor-IgG fusion protein (AdTNFRI) was injected intravenously into 6-week old transgenic mice, which resulted in high levels of TNFRI in both plasma and myocardium. AdTNFRI did not reverse cardiomegaly but abrogated myocardial inflammation. Furthermore, AdTNFRI blocked the myocardial expression of intercellular adhesion molecule-1 and downstream cytokines, including interleukin 1beta and monocyte chemotactic protein-1. Downregulation of alpha-myosin heavy chain was restored by the treatment, whereas upregulation of beta-myosin heavy chain was not reversed. In contrast, the downregulation of sarcoplasmic reticulum Ca(2+)-ATPase and phospholamban was normalized by AdTNFRI. Echocardiographic measurements showed that left ventricular end-systolic diameter was significantly larger in transgenic mice than in control mice, and this increase was reversed by the AdTNFRI treatment. However, left ventricular wall thickening was not reversed. CONCLUSIONS: These results suggest that anti-TNF therapy may hold promise in the treatment of end-stage heart failure. PMID- 10831528 TI - Coronary nitric oxide production in response to exercise and endothelium dependent agonists. AB - BACKGROUND: Endothelium-derived nitric oxide (NO) contributes to epicardial coronary artery vasodilation during exercise. However, blockade of NO production does not impair the increase in coronary blood flow (CBF) during exercise, suggesting that NO is not obligatory for exercise-induced coronary resistance vessel dilation. In contrast, the increases in CBF produced by endothelium dependent agonists are decreased after NO blockade. Consequently, this study was performed to determine whether the increase in coronary NO production in response to agonists is greater than that which occurs during exercise. METHODS AND RESULTS: We measured the oxidation products of NO (nitrate+nitrite=NO(x)) in aortic and coronary sinus plasma using chemiluminescence to assess NO(x) production across the coronary circulation in chronically instrumented dogs during a 3-stage treadmill exercise protocol and in response to intracoronary administration of the endothelium-dependent agonists acetylcholine (37.5 microg/min) and bradykinin (3.0 microg/min). No coronary NO(x) production could be detected at rest or during the first 2 stages of exercise; only at the highest level of exercise was a small increase in coronary NO(x) production measured. In contrast, coronary production of NO(x) was significantly increased in response to endothelium-dependent agonists. CONCLUSIONS: Coronary NO production in response to endothelium-dependent agonists is greater than in response to the increase in shear stress associated with exercise. These findings support previous studies suggesting that NO is not required for the coronary vasodilation that occurs in the normal heart during exercise. PMID- 10831529 TI - Prevention of rat cerebral aneurysm formation by inhibition of nitric oxide synthase. AB - BACKGROUND: Cerebral saccular aneurysm is a major cause of subarachnoid hemorrhage, one of the cerebrovascular diseases with the highest mortality. The mechanisms underlying the development of aneurysms, however, still remain unclear. We have made a series of reports on an animal model of experimentally induced cerebral aneurysms that resemble human cerebral aneurysms in their location and morphology, suggesting that the arterial wall degeneration associated with aneurysm formation develops near the apex of arterial bifurcation as a result of an increase in wall shear stress. Using the animal model and human specimens, we examined the role of nitric oxide (NO) in the degenerative changes and cerebral aneurysm formation. METHODS AND RESULTS: Inducible NO synthase (iNOS) was immunohistochemically located at the orifice of human and rat aneurysms. Nitrotyrosine distribution was also seen in the human aneurysm. Although no iNOS immunostaining was found in normal arteries, iNOS immunoreactivity was observed in parallel with the development of early aneurysmal changes in rats. In contrast, during the early development of aneurysm, endothelial NOS immunostaining in the endothelium was weakened compared with that in the control arteries. An NOS inhibitor, aminoguanidine, attenuated both early aneurysmal changes and the incidence of induced aneurysms. A defibrinogenic agent, batroxobin, which may diminish shear stress by reduction of blood viscosity, prevented iNOS induction as well as early aneurysmal changes. CONCLUSIONS: The evidence suggests that NO, particularly that derived from iNOS, is a key requirement for the development of cerebral aneurysm. The iNOS induction may be caused by an increase in shear stress near the apex. PMID- 10831530 TI - Effect of distension of the gallbladder on plasma renin activity in anesthetized pigs. AB - BACKGROUND: Gallbladder pathology has been associated with cardiovascular disease. Recently, we showed that gallbladder distension in anesthetized pigs reflexly increased heart rate, arterial pressure, and coronary and renal vascular resistance through efferent sympathetic mechanisms. Renin release is affected by sympathetic output, and angiotensin liberation may result in vasoconstriction. This study was undertaken to determine whether gallbladder distension primarily causes a reflex change in plasma renin activity (PRA) and to assess its influence on observed pressor and coronary responses as well as on regional vascular resistance. METHODS AND RESULTS: In 34 alpha-chloralose-anesthetized pigs, balloons positioned within the gallbladder were distended for 30 minutes with volumes of Ringer's solution equal to those of withdrawn bile. In 19 pigs, gallbladder distension at constant heart rate, arterial pressure, and renal flow increased PRA in the absence of changes in urinary sodium excretion. This increase was abolished by cervical vagotomy, section of renal nerves, or blockade of beta-adrenergic receptors. In another 15 pigs, blockade of angiotensin II receptors significantly attenuated the pressor and coronary, mesenteric, and iliac vasoconstriction responses to gallbladder distension. CONCLUSIONS: The present study showed that innocuous gallbladder distension primarily caused a reflex increase in PRA. This increase, which involved afferent vagal pathways and efferent sympathetic mechanisms related to beta-adrenergic receptors, contributed significantly to the pressor and coronary, mesenteric, and iliac vasoconstriction responses to gallbladder distension. PMID- 10831531 TI - Single low-dose administration of human recombinant hepatocyte growth factor attenuates intimal hyperplasia in a balloon-injured rabbit iliac artery model. AB - BACKGROUND: Previous studies have shown that repeated systemic administration of human recombinant hepatocyte growth factor (hrHGF) in mg/kg levels modulates the wound-healing process in various diseases. Recently, HGF has been characterized as one of the most potent endothelial-cell-specific growth factors. We tested our hypothesis that local delivery of hrHGF, even at low microg/kg levels (> or =2 orders of magnitude lower than systemically administered doses), might attenuate neointimal hyperplasia in response to vascular injury via accelerated reendothelialization. METHODS AND RESULTS: The iliac artery was denuded in 16 New Zealand White rabbits (3 kg), followed by administration, via a drug delivery catheter, of either hrHGF (10 microg; n = 11) or control vehicle (n=5) over 20 minutes. In pilot studies using this device, the drug permeated into the medial tissues, where it persisted for > or =24 hours. Four weeks after the local delivery of hrHGF, computer-assisted morphometric analysis revealed significant reduction in the intimal area (hrHGF, 0.37+/-0.21 versus control, 0.68+/-0.16 mm(2), mean +/- SD; P<0.05) but no change in the medial area (hrHGF, 1.03+/-0.21 versus control, 1.10+/-0.52 mm(2)). Scanning electron microscopy revealed extensive endothelialization with regular and confluent endothelial cell layer regeneration in the hrHGF-treated vessels. CONCLUSIONS: Accelerated endothelialization after local delivery of hrHGF, a novel and potent endothelial cell mitogen, effectively attenuates neointimal proliferation even after single low-dose administration. This observation could have potential therapeutic implications in the prevention of restenosis after angioplasty. PMID- 10831532 TI - Images in cardiovascular medicine. T-wave alternans preceding torsade de pointes ventricular tachycardia. PMID- 10831533 TI - Safety and effectiveness of dofetilide for conversion of atrial fibrillation and nesiritide for acute decompensation of heart failure: a report from the cardiovascular and renal advisory panel of the Food and Drug Administration. PMID- 10831534 TI - Three-dimensional densimetric analysis of coronary artery aneurysm. PMID- 10831535 TI - Endothelium-dependent and -independent perfusion reserve and the effect of L arginine on myocardial perfusion in patients with syndrome X. PMID- 10831536 TI - The G20210A prothrombin mutation and the Physicians' Health Study. PMID- 10831537 TI - Trends in antihypertensive drug treatment. PMID- 10831538 TI - Paradoxical embolism as a principal cause of stroke in atrial septal aneurysm. PMID- 10831539 TI - Treatment of acute myocardial infarction by primary coronary angioplasty or intravenous thrombolysis. PMID- 10831540 TI - Health care for all--in Maryland, at least. PMID- 10831541 TI - First Food and Drug Administration approval under humanitarian device exemption of a septal occluder for fenestrated fontan and muscular ventricular septal defects. PMID- 10831542 TI - Embryo implantation and GnRH antagonists: embryo implantation: the Rubicon for GnRH antagonists. AB - When gonadotrophin-releasing hormone (GnRH) was discovered, the agonist and antagonist of GnRH were developed to control the release of FSH and LH by the gonadotrophs. More than 10 years of research were needed to develop a GnRH antagonist free of histamine release. Recent studies have shown that these GnRH antagonists are effective in preventing a rise in LH during ovarian stimulation in IVF. However, a decrease in ongoing pregnancies seems to suggest that implantation rates per transferred embryo are reduced in GnRH antagonist stimulated cycles. In my opinion, these data highlight an area less well known to clinicians: the role of the GnRH antagonist at the cellular level in extrapituitary tissues. There are sufficient data in the literature suggesting that GnRH antagonist is an inhibitor of the cell cycle by decreasing the synthesis of growth factors. Given that, for folliculogenesis, blastomere formation and endometrium development, mitosis is everything; the interaction between the GnRH antagonist and the GnRH receptor (present in all these cells and tissues) may compromise the mitotic programme of these cells. This is the Rubicon for the GnRH antagonist: to demonstrate irrevocably that, at the minimal doses necessary to suppress LH release, it does not affect processes such as implantation, embryo development and folliculogenesis. PMID- 10831543 TI - Avoiding multiple pregnancies in ART: consideration of new strategies. AB - To maximize pregnancy rates, physicians who perform IVF, often transfer multiple embryos, which increases the multiple birth risk. Multiple birth infants are at significant risk for a number of adverse outcomes including preterm delivery, low birth weight, congenital malformations, fetal and infant deaths and long term morbidity and disability among survivors. Since the ultimate goal for an IVF treatment is the birth of a healthy infant, an important issue for all kinds of assisted reproductive technologies (ART) must be the reduction or elimination of multiple pregnancies. In this article, different strategies to avoid multiple births in ART are discussed. PMID- 10831544 TI - Does ICSI affect early serum beta-HCG in pregnancies achieved after IVF? AB - This study was conducted to compare early serum human chorionic gonadotrophin (HCG) concentrations in singleton pregnancies achieved after intracytoplasmic sperm injection (ICSI), with those achieved after conventional in-vitro fertilization (IVF). Early serum HCG, 14-16 days after embryo transfer, was analysed in 99 IVF pregnancies achieved after ICSI (group A), and compared to 105 conventional IVF pregnancies (group B). All women were treated at the IVF Unit, Lis Maternity Hospital. Records were studied retrospectively. The mean +/- SE serum HCG concentration on day 14 after embryo transfer in group A was 190.5 +/- 17.4 mIU/ml, compared to 195.7 +/- 14.03 mIU/ml in group B. HCG concentration 14 days after embryo transfer in both groups A and B was higher in women with mechanical factor than in couples with male factor infertility or unexplained infertility (246 +/- 31.4, 183.3 +/- 16.4, 177.98 +/- 14.3 mIU/ml respectively). On the 16th day after embryo transfer, the HCG concentration increased, and the difference between the groups was maintained. Only in the subgroup of unexplained infertility did we find a difference in concentrations of HCG between ICSI and conventional IVF: on the 16th day following embryo transfer in this group there was a significant difference in HCG concentrations (395. 8 +/- 21 and 545.6 +/- 45.7 respectively; P = 0.04). HCG concentrations did not differ overall in the conventional IVF pregnancies compared with those achieved by ICSI. However, a statistical difference in early serum HCG concentrations was found in relation to the aetiology of infertility. PMID- 10831545 TI - Effects of gonadotrophin-releasing hormone agonists on human ovarian steroid secretion in vivo and in vitro-results of a prospective, randomized in-vitro fertilization study. AB - The aim of this prospective randomized study was to compare the effects of two gonadotrophin-releasing hormone (GnRH) agonists, buserelin and triptorelin, on human ovarian follicular steroidogenesis, oocyte fertilization and IVF treatment outcome. Ovulatory, healthy women undergoing IVF were treated either with human menopausal gonadotrophin (HMG) alone or with HMG and one of the two GnRH agonists. Serum and follicular fluid hormonal concentrations and cultures of luteinizing granulosa cells obtained during follicular aspiration were analysed. GnRH agonist treatment significantly affected steroidogenesis both in serum and follicular fluid. In follicular fluid, progesterone and oestradiol concentrations were significantly elevated while testosterone concentrations were significantly lower in the triptorelin group. The ratios of testosterone/progesterone, oestradiol/progesterone but not oestradiol/testosterone concentrations were significantly affected by GnRH agonist administration. Similarly, the steroidogenic activity of luteinizing granulosa cells in vitro was significantly decreased in women treated with GnRH agonists. Women treated with GnRH agonists had significantly more fertilized oocytes and cleaving embryos. The results indicate a marked effect of GnRH agonists on the pattern of ovarian follicular steroidogenesis that cannot be explained solely by changes in gonadotrophin concentrations. PMID- 10831546 TI - Recombinant human FSH versus highly purified urinary FSH: a randomized study in intrauterine insemination with husbands' spermatozoa. AB - A randomized trial was carried out comparing recombinant FSH (rFSH) and highly purified urinary FSH (uFSH) in intrauterine insemination (IUI) with husbands' spermatozoa. A total of 45 women received rFSH (139 cycles), while 46 women received uFSH (155 cycles). The starting dose was 150 IU/day s.c., beginning on the second day, and on days 6-7 the dose was adjusted according to ovarian response, assessed by vaginal ultrasound and plasma oestradiol concentration. The pregnancy rate according to the intention to treat was 57.8% in rFSH versus 52.2% in uFSH, the corrected pregnancy rates 56.8% and 52.2%, and the cumulative pregnancy rates 69.6% and 61.0%, but the differences were not statistically significant. The per cycle pregnancy rate was 18.12% in rFSH and 15.48% in u-FSH, also not statistically significant. In the rFSH group, the consumption of FSH ampoules per cycle was significantly lower (19.20 +/- 7.02 versus 23. 80 +/- 10.78; P < 0.0001). The ratio of oestradiol/FSH ampoules was significantly higher in rFSH (56.45 +/- 31.26 versus 46.41 +/- 29. 25; P < 0.001). These data indicate that, in IUI cycles, rFSH has a higher potency than uFSH. PMID- 10831547 TI - Extracorporeal perfusion of the human uterus as an experimental model in gynaecology and reproductive medicine. AB - Experimental perfusion of various organs has primarily been used in transplantation medicine to study the physiology, pathophysiology and metabolism of tissues and cells. The purpose of this study was to establish an experimental model for the extracorporeal perfusion of the human uterus with recirculation of a modified, oxygenated Krebs-Henselait solution, in comparison with a non recirculating perfusion system. With consent of the patients we obtained 25 uteri after standard hysterectomy. We performed an isovolumetric exchange of the perfusion medium at different intervals from 1 to 6 h and examined pH, pO(2), pCO(2), lactate, lactate dehydrogenase and creatine kinase by taking arterial and venous samples every hour for 24 h. We found the perfusions to be adequate when maintaining flow rates at 15-35 ml/min and at pressures ranging from 70 to 130 mmHg. Isovolumetric exchange of the perfusate every 3-4 h was the maximum interval to keep pH, the arterio-venous gradients of pO(2) and pCO(2), and the other biochemical parameters in physiological ranges. Examination by light and electron microscopy showed well-preserved features of myometrial and endometrial tissue. However, a 6 h exchanging interval led to increasing hypoxic and cytolytic parameters during the whole perfusion period. X-ray studies using digital subtraction angiography and perfusion studies with methylene blue demonstrated the homogeneous distribution of the perfusion fluid throughout the entire organ. PMID- 10831548 TI - Growth hormone does not increase the expression of insulin-like growth factors and their receptor genes in the pre-menopausal human ovary. AB - A growing body of information now supports the existence of a complete intraovarian insulin-like growth factor I (IGF-I) system. Although the precise role of IGF-I in the context of ovarian physiology remains to be determined, it is likely that IGF-I may engage in the amplification of gonadotrophin hormonal action. These facts and experiments with animals establishing the ovaries of multiple species as a site of growth hormone (GH) reception and action have led to the use of recombinant GH (rGH) as an adjunctive agent to potentiate ovulation induction by exogenous gonadotrophins. Whether intraovarian IGF-I plays an intermediary role in GH hormonal action at the ovarian level remains uncertain at present. The aim of this study was to evaluate whether rGH administration to pre menopausal women could modify the expression of the IGF-I gene in the ovary. The expression of the IGF-I gene was examined in a time-dependent manner in normal pre-menopausal ovaries obtained from nine women treated with rGH and nine control women treated with placebo, using solution hybridization/RNase protection assays. Ovarian tissue samples were obtained 24 h (six women) and 7 days (12 women) following rGH/placebo injection. Total RNA (20 microg) from whole pre-menopausal ovaries (with or without rGH treatment) as well as from human granulosa cells was hybridized with a human IGF-I antisense RNA. IGF-I peptide, but not oestradiol, serum concentrations increased significantly 24 h after rGH injection. IGF-I gene, however, was not expressed in the luteinized granulosa cells and whole pre menopausal ovaries irrespectively of rGH treatment in ovarian samples analysed both 1 and 7 days following rGH injection. On the contrary, IGF-II mRNA transcribed from the fetal or fetal-neonatal IGF-II promoter and IGF-I receptor mRNA (both used as hybridization control) were both found in whole pre-menopausal ovary and luteinized granulosa cells. Nevertheless, no changes in the hybridization patterns were seen in the absence or presence of rGH. These studies demonstrate that rGH administration to normal premenopausal women does not change the expression of insulin-like growth factors and their receptor genes in the pre menopausal human ovary. Furthermore, these results provide further evidence against locally produced IGF-I as responsible for any ovarian effects seen in systemic rGH administration. PMID- 10831549 TI - Ovulation side and cycle fecundity: a retrospective analysis of frozen/thawed embryo transfer cycles. AB - The aim of the study was to evaluate a possible lateral difference in ovarian activity and its effect on cycle fecundity. A database was analysed retrospectively which covered 477 cycles in which frozen/thawed embryo transfer had been carried out. The cycles were spontaneous, with no hormonal treatment. Women with ovulation problems as a reason for infertility treatment were excluded. Factors investigated were the side of ovulation, endometrial thickness on cycle days 10-12 and on the day of embryo transfer, and pregnancy rate per embryo transfer. Ovulation was right-sided in 273 of the 477 cycles (57.2%) and left-sided in 204 of the cycles (42. 8%) (95% CI 38.3-47.2, P = 0.002). In the age category of 30-37 years, covering 288 cycles, the incidence of left-sided ovulation was 126 (43.7%, 95% CI 38.0-49.5, P = 0.034). In this category, the endometrial thickness (+/-SD) was significantly greater on the day of embryo transfer (i.e. at time of implantation) when there had been left-sided ovulation compared with right-sided [9.6 mm (2.0) versus 9.1 mm (1.8), P = 0.037]. In addition, the pregnancy rate per embryo transfer was higher when ovulation had been on the left side [32/126 (25.2%) versus right side 24/162 (14.8%), P = 0.035, 95% CI 0.0122-0.199]. In conclusion, right-sided ovulation was more frequent than left-sided in the whole group. This is the first study to report that the side of ovulation has a clinical impact. These data support the hypothesis that the side of ovulation is significant in terms of embryo implantation. PMID- 10831550 TI - Elevated expression of tumour necrosis factor alpha in cultured granulosa cells from women with endometriosis. AB - Fertilization and oocyte cleavage rates have previously been demonstrated to be lower for women with endometriosis undergoing IVF compared with controls. This might be related to impaired oocyte function, possibly due to an inflammatory milieu in the pelvis of these women, where an elevated concentration of many cytokines is documented. The aim of this study was to examine whether granulosa cells from women with endometriosis deviated with respect to production of the inflammatory cytokines interleukin-1beta, interleukin-6, interleukin-8 and tumour necrosis factor alpha (TNFalpha) compared with granulosa cells from healthy women, undergoing IVF for male infertility. The effect of human chorionic gonadotrophin on cytokine production was also investigated. Granulosa cells in follicular fluid were obtained at oocyte retrieval for IVF. Incubated cell culture media were analysed by enzyme-linked immunosorbent assay. The basal production of all four cytokines was higher in cells from women with endometriosis when compared to controls, although the increase was only significant for TNFalpha. Chorionic gonadotrophin had no significant effect, although it had a tendency to suppress cytokine release in both patient categories. Whether aberrant cytokine production in granulosa cells from women with endometriosis may disturb fertilizing capacity of oocytes requires study. PMID- 10831551 TI - Tumour necrosis factor alpha concentrations in the peritoneal fluid of infertile women with minimal or mild endometriosis are lower in patients with red lesions only than in patients without red lesions. AB - Tumour necrosis factor alpha (TNFalpha) of peritoneal fluid is believed to have important pro-inflammatory and angiogenic activities in the complex mechanisms of development of peritoneal endometriotic lesions. We have evaluated the concentrations of TNFalpha and macrophages in peritoneal fluid of infertile women with minimal or mild endometriosis and related them to the presence of peritoneal red lesions alone (red lesions only group; n = 11) or their absence (non-red lesions group; n = 36). A group of 39 infertile normo-ovulatory patients with normal pelvic anatomy was used as controls. TNFalpha concentrations did not differ between controls and either group of patients. Patients with red lesions only had significantly lower concentrations of TNFalpha in peritoneal fluid (P < 0.05) and had a higher proportion of samples with undetectable concentrations (P < 0.05) than patients without red lesions. The significant difference in TNFalpha concentrations was present when comparing the groups of patients in the proliferative phase but not in the secretory phase. Macrophage concentrations were not different in the groups. Our findings are compatible with an impairment of macrophage function and therefore lend support to the theory that an inappropriate immunological response of the peritoneal environment to regurgitated endometrium may play a part in the initial phases of endometriotic implants. PMID- 10831552 TI - CA125 production by the peritoneum: in-vitro and in-vivo studies. AB - The source of CA125 synthesis is still debated. Endometrial, peritoneal, ovarian and amniotic cells have been demonstrated to produce and secrete CA125. Different studies show that the peritoneum is a source of CA125. The present study aimed at investigating in vivo and in vitro the peritoneal contribution to circulating CA125. Cultures of uterine peritoneum, abdominal peritoneum and myometrium explants were performed and CA125 measured in the culture medium. To modulate the potential production of CA125, the explants were cultured with or without cycloheximide, bacterial lipopolysaccharide (LPS) or ascitic fluid. In a prospective study, we compared a group of patients after abdominal surgery (n = 19; nine men, 10 women) with a group after extra-abdominal surgery (n = 21; 11 men, 10 women), in order to detect a postoperative increase of serum CA125. De novo synthesis of CA125 could not be demonstrated in the cultures of uterine and abdominal peritoneum and in myometrium, but CA125 concentrations were detectable in the culture medium without being modulated by cycloheximide, LPS or ascitic fluid. After peritoneal surgery, the proportion of patients with increased serum CA125 was significantly higher (P < 0.03) after abdominal surgery as compared with extra-abdominal surgery. This is considered as indirect evidence for in-vivo production of CA125 by the peritoneum. PMID- 10831553 TI - Insulin sensitivity, insulin secretion, and metabolic and hormonal parameters in healthy women and women with polycystic ovarian syndrome. AB - To study the contributions of body mass, body fat distribution and family history of type 2 diabetes mellitus to hyperinsulinaemia, insulin secretion and resistance in polycystic ovarian syndrome (PCOS), 17 lean (LC) and 17 obese (OC) healthy control subjects, and 15 lean (LPCOS) and 28 obese (OPCOS) women with PCOS were investigated. Waist:hip ratio (WHR), serum concentrations of sex steroids, glucose and insulin during a 75 g oral glucose tolerance test (OGTT), and insulin and C-peptide early phase secretion, and insulin sensitivity index using a euglycaemic hyperinsulinaemic clamp were assessed. The PCOS subjects had a higher mean WHR than the controls. A trend towards hyperinsulinaemia and impairment of insulin sensitivity (including the rates of both glucose oxidation and non-oxidation) was observed in LPCOS subjects, but only in OPCOS subjects were these changes significant. Early phase insulin secretion but not the early phase C-peptide secretion was increased in PCOS subjects compared to controls, suggesting that peripheral hyperinsulinaemia in PCOS women was mainly due to the observed lowered hepatic insulin extraction and insulin resistance in skeletal muscle. Moreover, the presence of a family history of type 2 diabetes did not affect early phase insulin or C-peptide secretion in the PCOS group. These results confirm and strengthen earlier contentions, that insulin resistance is a characteristic defect in PCOS and is worsened particularly by abdominal obesity. PMID- 10831554 TI - Ovarian hyperstimulation syndrome and its effect on renal function in a renal transplant patient undergoing IVF treatment: case report. AB - Ovarian hyperstimulation syndrome (OHSS) in a renal transplant patient undergoing assisted conception treatment is reported. A couple with infertility secondary to tubal blockage and pelvic endometriosis received IVF treatment. Ovarian enlargement secondary to OHSS resulted in obstruction in the transplanted kidney and deterioration of renal function. No other systemic manifestations of OHSS were evident. Conservative management was successful and a twin live birth was later achieved by replacement of two frozen-thawed embryos. PMID- 10831556 TI - Experience with progesterone gel for luteal support in a highly successful IVF programme. AB - Efficacy of luteal support from single daily administration of Crinone((R)) 8% (progesterone gel) was tested in 43 women in an IVF programme with historical pregnancy rates >50%. Results were compared with those achieved in 46 women concurrently undergoing IVF and receiving 50 mg i.m. progesterone, and with historical data. Pregnancy rates (PR) were evaluated approximately 2 weeks after undergoing IVF by human chorionic gonadotrophin (HCG) measurement (total PR), by ultrasound 2-4 weeks later (clinical PR), and by counting births. Prior experience with other progesterone formulations was compared with that of Crinone 8%. Demographic and IVF characteristics were comparable for both concurrently treated groups. Total PR, clinical PR and live birth rates were similar for the Crinone and the concurrent i.m. progesterone groups: 31 (72.1%) versus 34 (73.9%); 26 (60.5%) versus 28 (60.9%), and 23 (53.5%) versus 23 (50%) respectively. Clinical PR and live birth rates were also similar to the last data reported to the Society for Assisted Reproduction Therapy. Overall acceptability of Crinone 8% was excellent. Among subjects with prior i.m. injection experience, most patients (69.2%) agreed that the gel was easier to use, less painful (76.9%) and less time-consuming (61.5%) than i.m. injections. In conclusion, Crinone 8% offers an appreciable improvement, as it provides an effective luteal support option that avoids painful i.m. injections. PMID- 10831555 TI - Should ICSI be the treatment of choice for all cases of in-vitro conception? AB - The objective of this study was to examine different clinical scenarios of in vitro conception, viz. fertilization with conventional IVF, IVF with high insemination concentration (HIC) and intracytoplasmic sperm injection (ICSI), and assess on a sibling oocyte comparison the hypothesis that ICSI should be performed in all cases requiring in-vitro conception. ICSI with husband's spermatozoa had a higher incidence of fertilization as compared with IVF or IVF with HIC with donor spermatozoa (if previous failure of fertilization had occurred) for unexplained infertility. Similarly, ICSI with husband's spermatozoa had as high an incidence of fertilization as IVF with donor spermatozoa for patients with severe oligozoospermia, asthenozoospermia and/or teratozoospermia, even when the spermatozoa were not selected for their morphology. Two studies were performed to assess ICSI in potential oocyte-related failure of IVF, viz. when fertilization occurred in >50% of oocytes for one group of patients, and in <50% of oocytes in a second group. In both of these studies a significant proportion of the oocytes that failed to fertilize with conventional IVF eventually fertilized after ICSI. The overall conclusion was that ICSI as a first option offers a higher incidence of fertilization, maximizes the number of embryos and minimizes the risk of complete failure of fertilization for all cases requiring in-vitro conception. However, among other concerns, current knowledge of ICSI as an outcome procedure does not provide the confidence to use this process in all cases of IVF for the time being. PMID- 10831557 TI - Increased frequency of mutations in DNA from infertile men with meiotic arrest. AB - In diverse organisms from yeast to mice, mutations in numerous genes required for DNA repair may lead to defects in meiosis. Although it is likely that meiosis is conserved throughout evolution, little is known about the genetics of meiosis in humans even though meiotic arrest associated with azoospermia is common. In this work, we compared the sequence fidelity of a polymorphic marker amplified from DNA of two groups of patients: those with testis biopsy suggesting meiotic arrest and those with normal spermatogenesis who were obstructed. We demonstrated that mutations are more common in DNA from testicular tissue derived from men with meiotic arrest than in DNA from testicular tissue derived from men with normal spermatogenesis and physical obstruction (P < 0.05). No mutations were observed in blood tissue from either group of men. This suggests the possibility that defects in genes required in DNA repair could contribute to meiotic arrest in men just as has been observed in other organisms. PMID- 10831558 TI - Follicular fluid concentrations of adrenomedullin, vascular endothelial growth factor and nitric oxide in IVF cycles: relationship to ovarian response. AB - Marked granulosa cell proliferation along with important changes in the vascular bed of the ovary characterize IVF cycles associated with multiple follicular growth and maturation. The present report investigated follicular fluid (FF) and circulating concentrations of adrenomedullin, vascular endothelial growth factor (VEGF) and nitric oxide (NO) in 70 IVF patients (14 of whom became pregnant); these three vasoactive substances may be implicated in extensive ovarian tissue remodelling. Serum and FF concentrations of oestradiol and progesterone were also measured in the 70 IVF cycles studied. Follicular fluid concentrations of VEGF and adrenomedullin but not nitrite/nitrate (the two stable oxidation products of NO metabolism) were significantly higher (P < 0.0001) than the corresponding circulating concentrations. Follicular fluid concentrations of oestradiol and progesterone were not correlated with those of adrenomedullin, VEGF or nitrite/nitrate. No relationship existed between circulating concentrations of adrenomedullin, VEGF or nitrite/nitrate on the day of oocyte aspiration and parameters of ovarian response to gonadotrophin stimulation. In contrast, FF adrenomedullin concentration showed a direct relationship with day 3 FSH serum concentration (r = 0.53, P < 0.01) and the number of ampoules of gonadotrophin administered (r = 0.36, P < 0.005), but an inverse correlation with the total number of oocytes retrieved (r = -0.29, P < 0.01) and the number of mature oocytes (r = -0.25, P < 0. 05). A positive correlation was found for FF VEGF concentration and chronological age (r = 0.29, P < 0.05) and ampoules of gonadotrophins administered (r = 0.30, P < 0.05). There was no relationship between nitrite/nitrate FF concentrations and parameters of ovarian response. Neither serum concentrations nor FF concentrations of adrenomedullin, VEGF or nitrite/nitrate were correlated with IVF outcome. This study suggested for the first time that increased FF concentrations of adrenomedullin can be a marker of decreased ovarian response in IVF. Our results also provide further evidence favouring an association between FF VEGF and patient's age, while on the basis of our findings NO measurements are not a useful marker of ovarian response. PMID- 10831559 TI - Restoration of a normal reproductive lifespan after grafting of cryopreserved mouse ovaries. AB - Fresh and frozen ovaries from 10 day old C57BL/6J-Gpi-1(a) mice were grafted orthotopically into ovariectomized B6CBF1 (homozygous Gpi-1(b)) recipients. The recipients were mated with B6CBF1 males. The birth and size of each litter was recorded. The electrophoretic variant of glucose phosphate isomerase was determined for each neonate. Twelve of 13 recipients of fresh ovary and 10 of 12 recipients of frozen ovary were fertile. Of these, 10 (fresh) and eight (frozen) had litters derived from the ovarian grafts only, or from the graft and native ovary. The breeding characteristics of recipients of fresh and frozen grafts were similar. The reproductive lifespan of the recipients of fresh (6.2 litters) and frozen (8.4 litters) grafts was similar to that of unoperated C57BL/6-Gpi-1(a) control females mated with B6CBF1 males (6.3 litters). Litter size was similar in recipients of grafted ovaries (fresh, 5.4 pups/litter; frozen, 6.3 pups/litter) and C57BL/6-Gpi-1(a) females (6.3 pups/litter). The results suggest that cryopreservation per se does not affect the long-term viability of ovarian tissue and provides an option for storing female germ cells. This is the first unequivocal demonstration that a normal reproductive lifespan can be restored by orthotopic grafting of frozen ovary. PMID- 10831560 TI - A prospective, randomized, controlled, double-blind, double-dummy comparison of recombinant and urinary HCG for inducing oocyte maturation and follicular luteinization in ovarian stimulation. AB - A randomized, controlled, double-blind, double-dummy, phase III clinical trial was conducted in 84 women to compare the efficacy of a s.c. injection of 250 microg recombinant human chorionic gonadotrophin (rHCG; Ovidrel) to an i.m. injection of 5000 IU urinary HCG (uHCG; Profasi) in inducing folliculogenesis, resumption of oocyte meiosis and luteinization after ovulation induction with recombinant follicle stimulating hormone (Gonal-F). The study primary endpoint was comparison of the number of oocytes retrieved per patient receiving either compound. Secondary comparisons included the number of oocytes retrieved per follicles aspirated; the number of mature oocytes; normally fertilized oocytes; and cleaved embryos. There were no statistically significant differences between groups for the primary endpoint (mean +/- SD oocytes retrieved 10.8 +/- 4.5 for rHCG versus 10.3 +/- 5.1 for uHCG) or each of the secondary endpoints except for increased concentrations of progesterone 6-7 days after rHCG administration (353.2 +/- 215.1 versus 234.1 +/- 129.4 nmol/l; P < 0. 004) and for HCG during the luteal phase following rHCG (P < 0.02). There were also no significant side effects for either drug. Since the confidence intervals for the difference of the number of oocytes retrieved between the two treatment groups were within the bounds defined by the multi-trial protocol equivalence between rHCG and uHCG could be declared. PMID- 10831561 TI - Fertility following ligation of internal iliac arteries for life-threatening obstetric haemorrhage: case report. AB - Bilateral ligation of internal iliac (hypogastric) arteries (BIL) is a life saving operation in cases of massive obstetric haemorrhage. This operation preserves reproductive function as opposed to the more commonly performed emergency hysterectomy in such situations. We report on effectiveness and future fertility in 12 women who had internal iliac ligation to control severe obstetric haemorrhage: in 10 out of the 12 women, BIL was successful. Of the two women who subsequently needed emergency hysterectomy, one woman died of disseminated intravascular coagulation. Of the eight women we were able to follow-up to assess reproductive performance, two did not desire future fertility. Three had subsequent pregnancies (50%), of whom two proceeded to term. We conclude that BIL is a safe and effective procedure for treating life-threatening obstetric haemorrhage with preservation of future fertility. This technique should be performed more often when indicated. PMID- 10831562 TI - Lack of association between smoking and DNA fragmentation in the spermatozoa of normal men. AB - Male factor infertility patients can have anomalies in their sperm nuclei, displaying high levels of loosely packaged chromatin and damaged DNA. The primary objectives of this study were to compare the extent of DNA fragmentation in the spermatozoa of healthy light and heavy smokers versus non-smokers, and to investigate its correlation with concentrations of the smoking markers cotinine and cadmium. A secondary objective was to compare the concentrations of blood cadmium and serum cotinine with corresponding concentrations in seminal plasma. Ninety-seven healthy male volunteers were divided into three groups: non-smokers, light and heavy smokers. There was no difference between the three groups with respect to age, number of ejaculations per week, serum testosterone concentration, and parameters of semen analysis. The percentages of DNA fragmentation in spermatozoa were not statistically different in the heavy smokers (12.11%), light smokers (11.66%) and non-smokers (20.41%). Serum and seminal plasma concentrations of cotinine were significantly higher in heavy smokers compared with the other groups (P < 0.0001). Median values for blood cadmium concentration were higher in heavy smokers (4.50 microg/l) than in light smokers (0.20 microg/l) and non-smokers (0.20 microg/l) (P < 0.001). Cadmium concentration in seminal plasma was significantly higher in heavy smokers (0.20 microg/l) than in light smokers (0.10 microg/l) and non-smokers (0. 10 microg/l) (P < 0.05). In summary, our results indicate no association between smoking and DNA fragmentation in the spermatozoa of healthy men. PMID- 10831563 TI - Objective evaluation of hyperactivated motility in rat spermatozoa using computer assisted sperm analysis. AB - The aim of this study was to use computer-assisted sperm analysis (CASA) to examine changes in motion parameters of rat spermatozoa incubated under culture conditions that support IVF. Rat cauda epididymal spermatozoa were evaluated in six replicate experiments, at 0 and 4h of incubation. CASA was conducted at 60 Hz on digital 1s tracks ( approximately 100 spermatozoa/rat). Mean values of CASA parameters that describe the vigour of spermatozoa [curvilinear velocity (VCL), amplitude of lateral head displacement (ALH) and beat cross frequency (BCF)] increased, while those indicating progressiveness [straight line velocity (VSL), linearity (LIN) and straightness (STR)] decreased between 0 and 4 h. Visual inspection of sperm tracks after 4 h of incubation revealed classical hyperactivation patterns. Bivariate models were evaluated to objectively define the subpopulation of hyperactivated (HA) spermatozoa. Of all models considered, ALH and LIN, VCL and LIN, BCF and LIN, VCL and BCF, and VCL and ALH showed significant changes in the percentage of HA spermatozoa after the 4 h incubation period. The efficacy of detecting HA spermatozoa was evaluated using sperm tracks that were visually classified as HA or progressive. VCL and LIN provided the most accurate prediction of HA spermatozoa. It was concluded that analysis of CASA data using bivariate models could be used to detect and monitor hyperactivation in rat spermatozoa. PMID- 10831564 TI - Leukocyte detection in human semen using flow cytometry. AB - This study set out to establish a new method, using flow cytometry, to evaluate leukocytes in semen. Ejaculates of 59 males, asymptomatic for genitourinary infections, were examined. Routine semen analyses were carried out as well as peroxidase and polymorphonuclear granulocyte-elastase detection. Leukocytes were detected combining flow cytometry and monoclonal antibodies (anti-CD45, anti CD53). This technique reliably assessed the total number of leukocytes and differentiated subpopulations even at low concentrations. The peroxidase test and elastase determination showed good specificity, but only moderate sensitivity versus flow cytometry combined with monoclonal antibodies. No significant association was observed between semen parameters and leukocytospermia whether evaluated by conventional methods or flow cytometry except for a moderate correlation between spermatozoa and CD53-positive cell concentrations. A first comparison of data from patients grouped on the basis of leukocytospermia (>10(6) white blood cells, WBC/ml) or non-leukocytospermia revealed no significant differences in semen parameters; lowering the threshold value for leukocytospermia to 2x10(5) WBC/ml, sperm concentration was reduced in the group with a low number of WBC identified by monoclonal antibodies. Flow cytometry using monoclonal antibodies was seen to be a simple, reproducible method that enables leukocytes in semen to be accurately detected and to identify WBC subpopulations without preliminary purification procedures. PMID- 10831565 TI - Analysis of DNA fragmentation, plasma membrane translocation of phosphatidylserine and oxidative stress in human spermatozoa. AB - The objectives of this cross-sectional observational study were: (i) to detect DNA damage and plasma membrane translocation of phosphatidylserine in purified sperm populations of high and low motility, and (ii) to analyse their relationship with the endogenous generation of reactive oxygen species. Ejaculates from infertile men were examined following gradient centrifugation. The main outcome measures were: sperm motion parameters (assessed with a computer analyser), generation of reactive oxygen species (measured by chemiluminescence), DNA damage (detected by terminal deoxynucleotidyl transferase-mediated dUDP nick end labelling and monoclonal antibody labelling of single-stranded DNA) and translocation of membrane phosphatidylserine (examined with annexin V staining). DNA fragmentation and membrane translocation of phosphatidyl-serine were observed in the fractions with low and high sperm motility in all patients. The fractions with low sperm motility had significantly higher proportion of cells with DNA damage and production of reactive oxygen species than the fractions with high sperm motility (P < 0.005). DNA fragmentation was significantly and positively correlated with the generation of reactive oxygen species (r = 0.42; P = 0.02). IN CONCLUSION: (i) spermatozoa from infertile men display translocation of membrane phosphatidylserine as diagnosed by annexin V positive staining; (ii) DNA damage (fragmentation and presence of single-stranded DNA) can be detected in ejaculated spermatozoa from infertile men in fractions with low and high sperm motility, and (iii) there is a relationship between DNA damage and oxidative stress. PMID- 10831566 TI - Angiotensin II in human seminal fluid. AB - The renin-angiotensin system (RAS) and angiotensin II are important in sperm function and male fertility. Angiotensin II type I (AT1) receptors have been identified in developing and ejaculated human spermatozoa, and angiotensin can stimulate sperm motility, the acrosome reaction and binding to the zona pellucida. However, there is little information on the availability of the hormone to spermatozoa during the reproductive process. Seminal plasma and blood plasma obtained from normal and subfertile subjects was extracted, and angiotensin content was analysed by radioimmunoassay. Values obtained for blood angiotensin II were within the normal range at 16.0 +/- 3.1 pg/ml (mean +/- SEM). Values for seminal plasma were usually 3-5 fold higher, at 51.6 +/- 9.3 pg/ml (n = 34, P < 0.0001). High performance liquid chromatography analysis showed that approximately 80% of the immunoreactive angiotensin was attributable to angiotensin II itself. However, seminal plasma angiotensin II concentrations were not correlated with blood angiotensin II, sperm concentration or sperm motility. The results show that immunoreactive angiotensin from a source other than the circulation is available to spermatozoa in human ejaculates. The results are consistent with the concept that angiotensin II has an important role in male fertility. PMID- 10831567 TI - In-vitro spermatogenesis resumption in men with maturation arrest: relationship with in-vivo blocking stage and serum FSH. AB - We have shown previously that germ cells recovered from some men with maturation arrest can resume spermatogenesis in vitro and give rise to late elongated spermatids. This study relates the ability of germ cells to differentiate in vitro to the stage at which spermatogenesis is blocked in vivo and to the patient's serum FSH concentration. The presence of germ cells at different stages of spermatogenesis was assessed, before and after culture, by classical cytology, by fluorescence in-situ hybridization and by immunocytochemistry with a germline specific marker. The proportion of cases of maturation arrest at the primary spermatocyte, secondary spermatocyte and spermatid stage in which in-vitro resumption of meiosis was achieved was 24.3% (9/37), 100% (3/3) and 51.1% (23/45) respectively. Serum FSH concentrations were higher than normal in most cases. However, lower values were measured in patients in whom in-vitro spermatogenesis was achieved compared with those in whom no progression was detected. These data show that, under the conditions of this study, germ cells from men with very high serum FSH concentrations (>20 IU/l) are less likely to resume spermatogenesis in vitro than those coming from men with only moderate increase (10-20 IU/l). PMID- 10831568 TI - Increase in scrotal temperature in car drivers. AB - Several recent studies have reported a gradual decline in sperm production in men. Endocrine disrupters as well as lifestyle have been suggested as risk factors. One lifestyle factor that may affect human fertility is driving a vehicle for a prolonged period. Several authors have suggested that driving position may increase the scrotal temperature. In order to validate this hypothesis we conducted continuous monitoring of scrotal temperature in real conditions, i.e. in men driving a car for a prolonged period. Nine volunteer men were asked to walk outside for 40 min and then to drive a car for 160 min. Scrotal temperatures were measured from thermocouples and values recorded every 2 min on a portable data recorder. Scrotal temperature increased significantly (P < 0.0001) in driving posture after 2 h of driving, reaching a value 1.7-2.2 degrees C higher than that recorded while walking. This link between driving position and increased scrotal temperature indicates a potential exposure of male reproductive function to lifestyle factors. PMID- 10831569 TI - Effects of five different barrier materials on postsurgical adhesion formation in the rat. AB - Postsurgical adhesion formation is a significant clinical problem within every surgical specialism. Due to the problems that adhesions cause, a wide variety of adjunctive treatments to prevent the formation and reformation of adhesions have been proposed. One of the modalities that has been studied extensively and that has been showing the most promising results is the so-called barrier method. The purpose of the present study was to compare the efficacy of five of these barrier materials in the prevention of postsurgical adhesion formation in a standardized rat adhesion model. It was concluded that no beneficial effect of Ringer's lactate on adhesion formation was seen. Significant reductions (P < 0.0001) in adhesion percentages compared to control animals were seen with Polyactive((TM)), PRECLUDE Peritoneal Membrane((TM)), Seprafilm((TM)) and Tissucol((TM)), but only PRECLUDE Peritoneal Membrane and Seprafilm significantly reduced adhesions (P < 0.01) when the barrier-treated peritoneal defects were compared with contralateral control-side peritoneal defects. The results of our study suggest that Seprafilm and PRECLUDE Peritoneal Membrane are superior to Tissucol and Polyactive in preventing adhesion formation. When Polyactive was still attached to the site of application during the second laparotomy, similar results to Seprafilm and PRECLUDE Peritoneal Membrane were seen. Future studies on the efficacy of a material to decrease adhesion formation should always include a comparison of several control materials in the same model. Our study indicates that Seprafilm or PRECLUDE Peritoneal Membrane might be used as standards of control. PMID- 10831570 TI - Surgical therapy in infertile men with ejaculatory duct obstruction: technique and outcome of a standardized surgical approach. AB - In severe oligozoospermia or azoospermia, low ejaculate volume, low ejaculate pH and little or no fructose in seminal plasma suggest an obstruction of the seminal pathways at the level of the prostate gland, when vasal aplasia and ejaculatory disorders are excluded. We report on our standardized surgical approach in 16 consecutive patients with this condition after clinical evaluation, semen analysis, endocrine assessment, testicular biopsy and transrectal ultrasonography. Pre-operatively, sperm analysis demonstrated typical low-volume ejaculates with azoospermia in 12 and severe oligozoospermia in four cases. Ultrasonography demonstrated seven central (Mullerian) and five lateral cystic lesions. Four cases with central obstruction revealed no ultrasonographic pathology. After intra-operative vasopuncture and vasography for definite localization of the level of obstruction, transurethral incision and/or resection of ejaculatory ducts (TURED) was performed. Patency was proven in 15 out of 16 cases by 'intra-operative chromotubation'. In nine out of 12 patients, spermatozoa could be harvested intra-operatively from the vas. During the follow up of 12 months, post-operative ejaculates showed persistent patency in six out of seven Mullerian cysts with concomitant improvement of sperm quality. Only three of the other nine cases remained patent with the worst results in lateral cystic lesions. Only two of the patients with Mullerian cysts have fathered a child so far. The data provide evidence for the effectiveness of surgical treatment of ejaculatory duct obstruction, especially in the case of central cystic lesions. The combination of surgery, cryostoring of spermatozoa retrieved intra-operatively and the possible storage of ejaculated spermatozoa post operatively creates the possibility of subsequently using reproductive techniques if pregnancy is not achieved. PMID- 10831571 TI - Microlaparoscopy for an intact ectopic pregnancy and endometriosis with the use of a diode laser: case report. AB - Microlaparoscopy is a development of endoscopic surgery which further reduces invasiveness of surgical procedures. The use of a diode laser in microlaparoscopy for the treatment of a patient with an intact ectopic pregnancy and endometriosis is described for the first time. As the diode laser has easy management and widely recognized precision, its use could be highly advantageous in such situations. The success achieved in this case contributes to the wider use of micro-endoscopic procedures. PMID- 10831572 TI - The position of the metaphase II spindle cannot be predicted by the location of the first polar body in the human oocyte. AB - When performing intracytoplasmic sperm injection (ICSI) on human oocytes, the injection is traditionally made at the 3 o'clock position, with the first polar body (PB) at the 12 or 6 o'clock position. This has been based on the assumption that the second meiotic metaphase II (MII) spindle lies in close proximity to the first PB. The objective of this study was to document the actual spatial relationship between the first PB and the MII spindle both in in-vivo matured (fresh) human MII oocytes and in oocytes matured in vitro. We found that the MII spindle was, on average, not located directly adjacent to the PB. The in-vivo group (n = 54) showed a mean deviation of the MII spindle from the position of the PB of 41. 7 degrees and the in-vitro group 26.6 degrees (n = 43). The difference between the angle of the two groups was statistically significant (P = 0.005), indicating that the lateral displacement of the first PB is only partly due to the denuding procedure during ICSI, because the in-vitro matured oocytes were denuded before extrusion of the first PB. The majority of the MII spindles in both groups were found in the same hemisphere as the first PB, suggesting that care should be taken to avoid damaging the MII spindle by inserting the ICSI needle in the other half of the oocyte. PMID- 10831573 TI - Progression to the blastocyst stage of embryos derived from testicular round spermatids. AB - Progression to the blastocyst stage of embryos derived from testicular round spermatids in men with non-obstructive azoospermia was studied. A total of 56 men were studied in whom partial spermatogenesis failure had occurred where only very few spermatozoa (fewer than the number of oocytes retrieved) were extracted from multiple testicular biopsy specimens. Oocytes remaining after intracytoplasmic injection of testicular spermatozoa (group 1) were injected with round spermatids (ROSI, group 2). Only embryos derived from group 1 were transferred. Remaining embryos were observed under culture for 8 days and their progression to the blastocyst stage was recorded. Of the 546 oocytes injected with testicular spermatozoa, 404 (73.9%) showed evidence of 2-pronuclear (2PN) fertilization. Injection of testicular round spermatids resulted in 2PN fertilization rate of 50% (P < 0.05). Using a four-point grading system, 53% of the good quality embryos (grade 1 or 2) in group 1 reached the blastocyst stage compared with 25% in group 2 (P < 0.05). The rate of progression to the blastocyst stage of grade 3 and grade 4 embryos was 46 and 8.5% in the two groups respectively (P < 0.05). Using a different three-point grading system for the blastocysts, 75.3% of the blastocysts in group 1 were either grade 1 or grade 2 and 24.7% were grade 3. However, in group 2 all blastocysts were grade 3. All embryos observed in group 1 reached the blastocyst stage by day 5 or 6 compared with 25% of the embryos reaching the blastocyst stage by this time in group 2. While 31.2% of the blastocysts in group 1 showed evidence of spontaneous hatching in vitro, none of the blastocysts in group 2 hatched. In conclusion, progression to the blastocyst stage occurred at a much lower and slower rate in embryos derived from testicular round spermatids. Furthermore, all blastocysts resulting from ROSI were of poor quality and none showed spontaneous hatching. These results may explain the dismal outcome associated with ROSI. PMID- 10831574 TI - Fertilization, pregnancy and embryo implantation rates after ICSI in cases of obstructive and non-obstructive azoospermia. AB - The aetiology of azoospermia can be grossly divided into obstructive and non obstructive causes. Although in both cases testicular spermatozoa can be used to treat male fertility, it is not well established whether success rates following intracytoplasmic sperm injection (ICSI) are comparable. Therefore, a retrospective analysis of fertilization, pregnancy and embryo implantation rates was performed following ICSI with testicular spermatozoa in obstructive or non obstructive azoospermia. In total, 193 ICSI cycles were carried out with freshly retrieved testicular spermatozoa; in 139 cases of obstructive and 54 cases of non obstructive azoospermia. The fertilization rate after ICSI with testicular spermatozoa in non-obstructive azoospermia was significantly lower than in obstructive azoospermia (67.8% versus 74.5%; P = 0.0167). Within the non obstructive group, the fertilization rate in the group of maturation arrest (47.0%) was significantly lower than in case of Sertoli cell-only (SCO) syndrome (71.2%) or germ cell hypoplasia (79. 5%). Embryo quality on day 2 after ICSI was similar for all groups. Pregnancy rates per transfer between obstructive (36.8%) and non-obstructive groups (36.7%) were similar. In cases of maturation arrest the pregnancy rate per transfer was lowest (20.0%) although not significantly different from SCO syndrome or hypoplasia groups. Embryo implantation rates were not different between the obstructive (19.6%) and non-obstructive groups (25.8%), and were lowest in cases of germ cell hypoplasia (15.8%). This retrospective analysis shows that although fertilization rate after ICSI with testicular spermatozoa in non-obstructive azoospermia is significantly lower than in obstructive azoospermia, pregnancy and embryo implantation rates are similar. PMID- 10831575 TI - The ability to generate normal Ca(2+) transients in response to spermatozoa develops during the final stages of oocyte growth and maturation. AB - Intracellular Ca(2+) oscillations at fertilization are responsible for triggering egg activation. The aim of this study was to examine the effect of the age of the oocyte donor and in-vitro maturation on the generation of Ca(2+) transients at fertilization. The results show that <10% of in-vivo and in-vitro matured oocytes from 19-day old mice develop to the blastocyst stage in vitro. In contrast, 43% of in-vivo and 25% of in-vitro matured oocytes from 24-day old mice developed to the blastocyst stage. In parallel experiments, intracellular Ca(2+) was monitored at fertilization. Oocytes from 19-day old mice generate significantly fewer transients than oocytes from 24-day old mice. In-vitro maturation significantly decreased the ability of oocytes from 19-day old mice but not 24-day old mice to generate Ca(2+) transients in response to spermatozoa. Furthermore, we investigated the effect of oocyte maturation on Ca(2+) signalling. Immature oocytes generated fewer Ca(2+) oscillations and ceased oscillating earlier than mature oocytes. These studies suggest that the ability to generate Ca(2+) transients in response to spermatozoa increases in the final stages of oocyte development and during oocyte maturation. This may contribute to the acquisition of developmental competence in the final stages of oogenesis. PMID- 10831576 TI - Embryos with high implantation potential after intracytoplasmic sperm injection can be recognized by a simple, non-invasive examination of pronuclear morphology. AB - Embryos are conventionally selected for transfer based on the evaluation of the cleavage speed and extent of blastomere fragmentation. Here we examined whether the predictive value of these criteria, as indicators of the chance of embryo implantation, can be further potentiated by adding previously described criteria reflecting the regularity of pronuclear development. In a group of embryos selected for transfer in 380 fresh embryo transfer cycles according to the conventional criteria, the transfer of only those embryos that developed from zygotes judged normal at the pronuclear stage (pattern 0) gave significantly higher pregnancy (44.8%) and implantation (30.2%) rates compared with the pregnancy (22.1%; P < 0. 05) and implantation rates (11.2%; P < 0.001) for the transfers of only those embryos that developed from zygotes judged abnormal (non pattern 0). The transfer of only one pattern 0 embryo was sufficient for the optimal chance of pregnancy (no differences in pregnancy rates after transfer of one, two or three pattern 0 embryos), whereas the transfer of two pattern 0 embryos mostly resulted in a twin pregnancy. The inclusion of the criteria based on pronuclear morphology can thus lead to the application of a single embryo transfer policy and optimize the selection of embryos for transfer and cryopreservation. PMID- 10831577 TI - Is there a human fetal gallbladder contractility during pregnancy? AB - The purpose of this study was to evaluate whether human fetal gallbladder contractility exists in the second half of pregnancy. Ultrasound examinations were performed on 54 normal pregnant women from 20 to 40 weeks of gestation. Fetal gallbladder volume was monitored every 30 min from 08:00 to 18:00 h in each patient. Maximum gallbladder volume was related linearly with gestational age between 20 and 32-35 weeks of gestation, after which a plateau was observed. Minimum gallbladder volume was unchanged throughout gestation. Functional capacity (maximum volume - minimum volume) of the fetal gallbladder increased linearly with advancing gestation until 32-35 weeks gestation, and thereafter was constant. Contractility rate [(maximum volume - minimum volume/maximum volume)x100] increased curvilinearly with advancing gestation (R(2) = 30.7%, P < 0.0001). The daily change in fetal gallbladder volume showed a typical sinusoidal pattern, and the contractility cycle of gallbladder volume was unchanged during pregnancy (3.1 +/- 0.6 h). These results suggest that there is an apparent gallbladder contractility in human fetuses in utero, and that maternal meals seem not to affect the volume of the fetal gallbladder. Further study is needed to clarify the physiological role of fetal gallbladder contractility during pregnancy. PMID- 10831578 TI - Antenatal dexamethasone and the growth hormone-insulin-like growth factor axis. AB - Dexamethasone administration has marked effects on the growth hormone-insulin like growth factor axis (GH-IGF) in animal and human studies. During pregnancy in the rat, it is associated with fetal growth restriction due to inhibition of IGF bioactivity. In the human only repeated dosages have been associated with fetal growth restriction. The aim of this study is to test the hypothesis that antenatal dexamethasone administration to pregnant women is associated with reduced activity of the GH-IGF axis. To achieve this blood samples were taken from 12 pregnant women pre- and at 24 h and 48 h after dexamethasone administration. In these samples GH, IGF-I, IGF bioactivity and IGF binding protein (IGFBP)-3 protease activity were measured. In view of the interaction between insulin and the GH-IGF axis, glucose and insulin concentrations were also measured. There were no significant differences between the concentrations of GH, IGF-I, IGF bioactivity and IGFBP-3 protease activity before and after dexamethasone. The concentrations of glucose and insulin were significantly higher at 24 h, but not 48 h post-dexamethasone. It is concluded that a single antenatal course of dexamethasone does not alter the GH-IGF-I axis in pregnant women at the time points studied. PMID- 10831579 TI - Intrauterine sonographic measurement of embryonic brain vesicle. AB - Our purpose was to evaluate embryonic brain vesicle measurements using intrauterine sonography in early first-trimester pregnancy. Fifty-one women about to undergo therapeutic abortion from 7-9.9 weeks gestational age were studied with a specially developed flexible catheter-based high-resolution real-time miniature (2.4 mm in outer diameter) ultrasound transducer (20 MHz). Length, width and height of telencephalon, diencephalon, mesencephalon and rhombencephalon were measured. The normal range of embryonic brain vesicle measurements for each day of pregnancy was determined. Curvilinear relationships were found between the menstrual age and telencephalon height (r(2) = 71.2%), diencephalon length (r(2) = 39. 6%), width (r(2) = 39.4%) and height (r(2) = 48.3%) and mesencephalon length (r(2) = 59.0%) respectively. Linear relationships were found between the menstrual age and telencephalon width (r(2) = 41.4%), mesencephalon height (r(2) = 58.7%) and rhombencephalon length (r(2) = 44.9%), width (r(2) = 56.8%) and height (r(2) = 35.5%) respectively. Telencephalon length and mesencephalon width were constant throughout menstrual age. These results suggest that intrauterine sonography provides accurate embryonic brain measurements in utero. Moreover, intrauterine sonography may become an important modality in future embryological research and in detection of embryonic brain developmental disorders in early first-trimester pregnancy. PMID- 10831580 TI - First trimester diagnosis of conjoined twins in a triplet pregnancy after IVF and ICSI: case report. AB - A case of conjoined twins in a triplet pregnancy after in-vitro fertilization and intracytoplasmic sperm injection is described. The diagnosis was made by high resolution vaginal sonography, as early as the eighth week of gestation. The third fetus, of different chorionicity, was normal. Selective termination was successfully done at 12 weeks. The possible association between assisted reproduction and conjoined twinning is discussed. The importance of expert early vaginal sonography of pregnancy resulting from assisted reproduction technology is emphasized. PMID- 10831581 TI - Testicular tissue cryopreservation in boys. Ethical and legal issues: case report. AB - Sperm preservation prior to chemotherapy and radiotherapy is common practice in adult males. Spermatozoa are usually retrieved from an ejaculated sample although there are occasions when testicular tissue is used as the source. These techniques of sperm preservation present minimal ethical objections as the patients give their informed consent. Sperm preservation in children presents practical and ethical dilemmas in that the children cannot always give their informed consent, there are no regulatory guidelines and there is no guarantee that spermatogenesis is occurring. With the rapid advances in reproductive technology and the possible future use of immature germ cells by in-vitro maturation or transplantation, the demand for immature testicular tissue preservation is likely to increase. More information for the parents and oncologists with regard to this subject is needed to allow informed decisions to be made on behalf of the children. These issues are discussed using two cases of children having testicular tissue preservation. PMID- 10831583 TI - The association between the sexes of children within sibships. PMID- 10831582 TI - Long term follow-up of children born after inadvertent administration of a GnRH analogue in early pregnancy. PMID- 10831584 TI - Ovarian reserve: A simple mathematical problem? PMID- 10831585 TI - Thermal balloon ablation versus endometrial resection for treatment of abnormal uterine bleeding. PMID- 10831587 TI - Fluorescence properties and functional roles of tryptophan residues 60d, 96, 148, 207, and 215 of thrombin. AB - Conservative Trp-to-Phe mutations were individually created in human thrombin at positions 60d, 96, 148, 207, and 215. Fluorescence intensities for these residues varied by a factor of 6. Residues 60d, 96, 148, and 215 transferred energy to the thrombin inhibitor 5-dimethylaminonaphthalene-1-sulfonylarginine-N-(3-ethyl-1,5- pentanediyl)amide efficiently, but residue 207 did not. Intensities correlated inversely with exposure to solvent, and measured and theoretical energy transfer efficiencies agreed well. Function was measured with respect to fibrinogen clotting, platelet and factor V activation, inhibition by antithrombin, and the thrombomodulin-dependent activation of protein C and thrombin-activable fibrinolysis inhibitor (TAFI). All activities of W96F and W207F ranged from 74 to 154% of the wild-type activity. This was also true for W148F, except for inhibition by antithrombin, where it showed 60% activity. W60dF was deficient by 30, 57, and 43% with fibrinogen clotting, platelet activation, and factor V cleavage (Arg(1006)), respectively. W215F was deficient by 90, 55, and 56% with fibrinogen clotting, platelet activation, and factor V cleavage (Arg(1536)). With protein C and TAFI, W96F, W148F, and W207F were normal. W60dF, however, was 76 and 23% of normal levels with protein C and TAFI, respectively. In contrast, W215F was 25 and 124% of normal levels in these reactions. Thus, many activities of thrombin are retained upon substitution of Trp with Phe at positions 96, 148, and 207. Trp(60d), however, appears to be very important for TAFI activation, and Trp(215) appears to very important for clotting and protein C activation. PMID- 10831586 TI - Phosphorylation at serine 10, a major phosphorylation site of p27(Kip1), increases its protein stability. AB - The association of the p27(Kip1) protein with cyclin and cyclin-dependent kinase complexes inhibits their kinase activities and contributes to the control of cell proliferation. The p27(Kip1) protein has now been shown to be phosphorylated in vivo, and this phosphorylation reduces the electrophoretic mobility of the protein. Substitution of Ser(10) with Ala (S10A) markedly reduced the extent of p27(Kip1) phosphorylation and prevented the shift in electrophoretic mobility. Phosphopeptide mapping and phosphoamino acid analysis revealed that phosphorylation at Ser(10) accounted for approximately 70% of the total phosphorylation of p27(Kip1), and the extent of phosphorylation at this site was approximately 25- and 75-fold greater than that at Ser(178) and Thr(187), respectively. The phosphorylation of p27(Kip1) was markedly reduced when the positions of Ser(10) and Pro(11) were reversed, suggesting that a proline directed kinase is responsible for the phosphorylation of Ser(10). The extent of Ser(10) phosphorylation was markedly increased in cells in the G(0)-G(1) phase of the cell cycle compared with that apparent for cells in S or M phase. The p27(Kip1) protein phosphorylated at Ser(10) was significantly more stable than the unphosphorylated form. Furthermore, a mutant p27(Kip1) in which Ser(10) was replaced with glutamic acid in order to mimic the effect of Ser(10) phosphorylation exhibited a marked increase in stability both in vivo and in vitro compared with the wild-type or S10A mutant proteins. These results suggest that Ser(10) is the major site of phosphorylation of p27(Kip1) and that phosphorylation at this site, like that at Thr(187), contributes to regulation of p27(Kip1) stability. PMID- 10831588 TI - Functional and structural analysis of ClC-K chloride channels involved in renal disease. AB - ClC-K channels belong to the CLC family of chloride channels and are predominantly expressed in the kidney. Genetic evidence suggests their involvement in transepithelial transport of chloride in distal nephron segments; ClC-K1 gene deletion leads to nephrogenic diabetes insipidus in mice, and mutations of the hClC-Kb gene cause Bartter's syndrome type III in humans. Expression of rClC-K1 in Xenopus oocytes yielded voltage-independent currents that were pH-sensitive, had a Br(-) > NO(3)(-) = Cl(-) > I(-) conductance sequence, and were activated by extracellular calcium. A glutamate for valine exchange at amino acid position 166 induced strong voltage dependence and altered the conductance sequence of ClC-K1. This demonstrates that rClC-K1 indeed functions as an anion channel. By contrast, we did not detect currents upon hClC Kb expression in Xenopus oocytes. Using a chimeric approach, we defined a protein domain that, when replaced by that of rClC-K1, allowed the functional expression of a chimera consisting predominantly of hClC-Kb. Its currents were linear and were inhibited by extracellular acidification. Contrasting with rClC-K1, they displayed a Cl(-) > Br(-)> I(-) > NO(3)(-) conductance sequence and were not augmented by extracellular calcium. Insertion of point mutations associated with Bartter's syndrome type III destroyed channel activity. We conclude that ClC-K proteins form constitutively open chloride channels with distinct physiological characteristics. PMID- 10831589 TI - Factor VIII C2 domain contains the thrombin-binding site responsible for thrombin catalyzed cleavage at Arg1689. AB - Thrombin-catalyzed factor VIII activation is an essential positive feedback mechanism regulating intrinsic blood coagulation. A factor VIII human antibody, A FF, with C2 epitope, exclusively inhibited factor VIII activation and cleavage at Arg(1689) by thrombin. The results suggested that A-FF prevented the interaction of thrombin with factor VIII and that the C2 domain was involved in the interaction with thrombin. We performed direct binding assays using anhydro thrombin, a catalytically inactive derivative of thrombin in which the active site serine is converted to dehydroalanine. Intact factor VIII, 80-kDa light chain, 72-kDa light chain, and heavy chain fragments bound dose-dependently to anhydro-thrombin, and the K(d) values were 48, 150, 106, and 180 nm, respectively. The C2 and A2 domains also dose-dependently bound to anhydro thrombin, and the K(d) values were 440 and 488 nm, respectively. The A1 domain did not bind to anhydro-thrombin. A-FF completely inhibited C2 domain binding to anhydro-thrombin (IC(50), 18 nm), whereas it did not inhibit A2 domain binding. Furthermore, C2-specific affinity purified F(ab)'(2) of A-FF, and the recombinant C2 domain inhibited thrombin cleavage at Arg(1689). Our results indicate that the C2 domain contains the thrombin-binding site responsible for the cleavage at Arg(1689). PMID- 10831590 TI - Sites in the A2 subunit involved in the interfactor VIIIa interaction. AB - Factor VIIIa is a trimer of the A1, A2, and A3-C1-C2 subunits. Regions in the A2 subunit that interact with the A1/A3-C1-C2 dimer were localized using synthetic peptides derived from A2 sequences showing high probability of being surface exposed. Peptides were restricted to residues 373-562 of A2 based on the earlier observation that this region of A2 reacts with A1 using a zero length cross linker. Peptides were assessed for their capacity to inhibit the reconstitution of factor VIIIa from the isolated A1/A3-C1-C2 dimer and A2 subunit. Reconstitution was monitored using both regeneration of factor VIIIa activity and fluorescence quenching of an acrylodan-labeled A2 (Ac-A2) by fluorescein-labeled A1/A3-C1-C2. The activity assay identified four peptides as inhibitors, residues 373-395 (IC(50) = 65 micrometer), 418-428 (IC(50) = 25 micrometer), 482-493 (IC(50) = 325 micrometer), and 518-533 (IC(50) = 585 micrometer). The 373-395 and 518-533 peptides eliminated the fluorescence quenching of Ac-A2, whereas the 418 428 peptide reduced but did not eliminate Ac-A2 quenching. Peptide 482-493 had no effect on the fluorescence quenching of Ac-A2 suggesting that the peptide did not directly affect reassociation of the factor VIIIa subunits. These results identify three regions in the A2 subunit (373-395, 418-428, and 518-533) that interact with the A1/A3-C1-C2 dimer. Furthermore, comparison of results obtained using the two assays distinguish inhibition of the intersubunit interactions from intermolecular interactions. PMID- 10831591 TI - Regulation of protein 4.1R, p55, and glycophorin C ternary complex in human erythrocyte membrane. AB - Three binary protein-protein interactions, glycophorin C (GPC)-4.1R, GPC-p55, and p55-4.1R, constitute the GPC-4.1R-p55 ternary complex in the erythrocyte membrane. Little is known regarding the molecular basis for the interaction of 4.1R with either GPC or p55 and regarding the role of 4.1R in regulating the various protein-protein interactions that constitute the GPC-4.1R-p55 ternary complex. In the present study, we present evidence that sequences in the 30-kDa domain encoded by exon 8 and exon 10 of 4.1R constitute the binding interfaces for GPC and p55, respectively. We further show that 4.1R increases the affinity of p55 binding to GPC by an order of magnitude, implying that 4.1R modulates the interaction between p55 and GPC. Finally, we document that binding of calmodulin to 4.1R decreases the affinity of 4.1R interactions with both p55 and GPC in a Ca(2+)-dependent manner, implying that the GPC-4.1R-p55 ternary protein complex can undergo dynamic regulation in the erythrocyte membrane. Taken together, these findings have enabled us to identify an important role for 4.1R in regulating the GPC-4.1R-p55 ternary complex in the erythrocyte membrane. PMID- 10831592 TI - Localization of disulfide bonds in the cystine knot domain of human von Willebrand factor. AB - von Willebrand factor (VWF) is a multimeric glycoprotein that is required for normal hemostasis. After translocation into the endoplasmic reticulum, proVWF subunits dimerize through disulfide bonds between their C-terminal cystine knot like (CK) domains. CK domains are characterized by six conserved cysteines. Disulfide bonds between cysteines 2 and 5 and between cysteines 3 and 6 define a ring that is penetrated by a disulfide bond between cysteines 1 and 4. Dimerization often is mediated by additional cysteines that differ among CK domain subfamilies. When expressed in a baculovirus system, recombinant VWF CK domains (residues 1957-2050) were secreted as dimers that were converted to monomers by selective reduction and alkylation of three unconserved cysteine residues: Cys(2008), Cys(2010), and Cys(2048). By partial reduction and alkylation, chemical and proteolytic digestion, mass spectrometry, and amino acid sequencing, the remaining intrachain disulfide bonds were characterized: Cys(1961)-Cys(2011) (), Cys(1987)-Cys(2041) (), Cys(1991)-Cys(2043) (), and Cys(1976)-Cys(2025). The mutation C2008A or C2010A prevented dimerization, whereas the mutation C2048A did not. Symmetry considerations and molecular modeling based on the structure of transforming growth factor-beta suggest that one or three of residues Cys(2008), Cys(2010), and Cys(2048) in each subunit mediate the covalent dimerization of proVWF. PMID- 10831593 TI - Cellular localization of membrane-type serine protease 1 and identification of protease-activated receptor-2 and single-chain urokinase-type plasminogen activator as substrates. AB - Membrane-type serine protease 1 (MT-SP1) was recently cloned, and we now report its biochemical characterization. MT-SP1 is predicted to be a type II transmembrane protein with an extracellular protease domain. This localization was experimentally verified using immunofluorescent microscopy and a cell-surface biotinylation technique. The substrate specificity of MT-SP1 was determined using a positional scanning-synthetic combinatorial library and substrate phage techniques. The preferred cleavage sequences were found to be (P4-(Arg/Lys)P3 (X)P2-(Ser)P1-(Arg)P1'-(Ala)) and (P4-(X)P3-(Arg/Lys)P2-(Ser)P1(Arg) P1'(Ala)), where X is a non-basic amino acid. Protease-activated receptor 2 (PAR2) and single-chain urokinase-type plasminogen activator are proteins that are localized to the extracellular surface and contain the preferred MT-SP1 cleavage sequence. The ability of MT-SP1 to activate PARs was assessed by exposing PAR-expressing Xenopus oocytes to the soluble MT-SP1 protease domain. The latter triggered calcium signaling in PAR2-expressing oocytes at 10 nm but failed to trigger calcium signaling in oocytes expressing PAR1, PAR3, or PAR4 at 100 nm. Single chain urokinase-type plasminogen activator was activated using catalytic amounts of MT-SP1 (1 nm), but plasminogen was not cleaved under similar conditions. The membrane localization of MT-SP1 and its affinity for these key extracellular substrates suggests a role of the proteolytic activity in regulatory events. PMID- 10831594 TI - Protein kinase C [micro] is regulated by the multifunctional chaperon protein p32. AB - We identified the multifunctional chaperon protein p32 as a protein kinase C (PKC)-binding protein interacting with PKCalpha, PKCzeta, PKCdelta, and PKC mu. We have analyzed the interaction of PKC mu with p32 in detail, and we show here in vivo association of PKC mu, as revealed from yeast two-hybrid analysis, precipitation assays using glutathione S-transferase fusion proteins, and reciprocal coimmunoprecipitation. In SKW 6.4 cells, PKC mu is constitutively associated with p32 at mitochondrial membranes, evident from colocalization with cytochrome c. p32 interacts with PKC mu in a compartment-specific manner, as it can be coimmunoprecipitated mainly from the particulate and not from the soluble fraction, despite the presence of p32 in both fractions. Although p32 binds to the kinase domain of PKC mu, it does not serve as a substrate. Interestingly, PKC mu-p32 immunocomplexes precipitated from the particulate fraction of two distinct cell lines, SKW 6.4 and 293T, show no detectable substrate phosphorylation. In support of a kinase regulatory function of p32, addition of p32 to in vitro kinase assays blocked, in a dose-dependent manner, aldolase but not autophosphorylation of PKC mu, suggesting a steric hindrance of substrate within the kinase domain. Together, these findings identify p32 as a novel, compartment specific regulator of PKC mu kinase activity. PMID- 10831595 TI - Ligands presumed to label high affinity and low affinity ATP binding sites do not interact in an (alpha beta)2 diprotomer in duck nasal gland Na+,K+-ATPase, nor Do the sites coexist in native enzyme. AB - The interaction of ligands deemed to be ATP analogues with renal Na(+),K(+) ATPase suggests that two ATP binding sites coexist on each functional unit. Previous studies in which fluorescein 5-isothiocyanate (FITC) was used to label the high affinity ATP site and 2'(3')-O-(2,4,6-trinitrophenyl)adenosine 5' diphosphate (TNP-ADP) was used to probe the low affinity site suggested that the two sites coexist on the same alphabeta protomer. Other studies in which FITC labeled the high affinity site and erythrosin-5-isothiocyanate (ErITC) labeled the low affinity site led to the conclusion that the high and low affinity sites exist on separate interacting protomers in a functional diprotomer. We report here that at 100% inhibition of ATPase activity by FITC, each alphabeta protomer of duck nasal gland enzyme has a single bound FITC. Both TNP-ADP and ErITC interact with FITC-bound protomers, which unambiguously demonstrates that putative high and low affinity ATP sites coexist on the same protomer. In unlabeled nasal gland enzyme, TNP-ADP and ErITC inhibit both ATPase activity and p-nitrophenyl phosphatase activity, functions attributed to the putative high and low affinity ATP site, respectively, by interacting with a single site with characteristics of the high affinity ATP binding site. In FITC-labeled enzyme, TNP-ADP and ErITC inhibit p- nitrophenyl phosphatase activity but at much higher concentrations than with the unmodified enzyme. Low affinity sites do not exist on the unmodified enzyme but can be detected only after the high affinity site is modified by FITC. PMID- 10831596 TI - Severe impairment of salivation in Na+/K+/2Cl- cotransporter (NKCC1)-deficient mice. AB - The salivary fluid secretory mechanism is thought to require Na(+)/K(+)/2Cl(-) cotransporter-mediated Cl(-) uptake. To directly test this possibility we studied the in vivo and in vitro functioning of acinar cells from the parotid glands of mice with targeted disruption of Na(+)/K(+)/2Cl(-) cotransporter isoform 1 (Nkcc1), the gene encoding the salivary Na(+)/K(+)/2Cl(-) cotransporter. In wild type mice NKCC1 was localized to the basolateral membranes of parotid acinar cells, whereas expression was not detected in duct cells. The lack of functional NKCC1 resulted in a dramatic reduction (>60%) in the volume of saliva secreted in response to a muscarinic agonist, the primary in situ salivation signal. Consistent with defective Cl(-) uptake, a loss of bumetanide-sensitive Cl(-) influx was observed in parotid acinar cells from mice lacking NKCC1. Cl(-)/ HCO(3)(-) exchanger activity was increased in parotid acinar cells isolated from knockout mice suggesting that the residual saliva secreted by mice lacking NKCC1 is associated with anion exchanger-dependent Cl(-) uptake. Indeed, expression of the Cl(-)/ HCO(3)(-) exchanger AE2 was enhanced suggesting that this transporter compensates for the loss of functional Na(+)/K(+)/2Cl(-) cotransporter. Furthermore, the ability of the parotid gland to conserve NaCl was abolished in NKCC1-deficient mice. This deficit was not associated with changes in the morphology of the ducts, but transcript levels for the alpha-, beta-, and gamma subunits of the epithelial Na(+) channel were reduced. These data directly demonstrate that NKCC1 is the major Cl(-) uptake mechanism across the basolateral membrane of acinar cells and is critical for driving saliva secretion in vivo. PMID- 10831597 TI - Modulation of the oligomerization state of the bovine F1-ATPase inhibitor protein, IF1, by pH. AB - Bovine IF(1), a basic protein of 84 amino acids, is involved in the regulation of the catalytic activity of the F(1) domain of ATP synthase. At pH 6.5, but not at basic pH values, it inhibits the ATP hydrolase activity of the enzyme. The oligomeric state of bovine IF(1) has been investigated at various pH values by sedimentation equilibrium analytical ultracentrifugation and by covalent cross linking. Both techniques confirm that the protein forms a tetramer at pH 8, and below pH 6.5, the protein is predominantly dimeric. By covalent cross-linking, it has been found that at pH 8.0 the fragment of IF(1) consisting of residues 44-84 forms a dimer, whereas the fragment from residues 32-84 is tetrameric. Therefore, some or all of the residues between positions 32 and 43 are necessary for tetramer formation and are involved in the pH-sensitive interconversion between dimer and tetramer. One important residue in the interconversion is histidine 49. Mutation of this residue to lysine abolishes the pH-dependent activation inactivation, and the mutant protein is active and dimeric at all pH values investigated. It is likely from NMR studies that the inhibitor protein dimerizes by forming an antiparallel alpha-helical coiled-coil over its C-terminal region and that at high pH values, where the protein is tetrameric, the inhibitory regions are masked. The mutation of histidine 49 to lysine is predicted to abolish coiled-coil formation over residues 32-43 preventing interaction between two dimers, forcing the equilibrium toward the dimeric state, thereby freeing the N-terminal inhibitory regions and allowing them to interact with F(1). PMID- 10831598 TI - Conserved walker A Ser residues in the catalytic sites of P-glycoprotein are critical for catalysis and involved primarily at the transition state step. AB - P-glycoprotein mutants S430A/T and S1073A/T, affecting conserved Walker A Ser residues, were characterized to elucidate molecular roles of the Ser and functioning of the two P-glycoprotein catalytic sites. Results showed the Ser-OH is critical for MgATPase activity and formation of the normal transition state, although not for initial MgATP binding. Mutation to Ala in either catalytic site abolished MgATPase and transition state formation in both sites, whereas Thr mutants had similar MgATPase to wild-type. Trapping of 1 mol of MgADP/mol of P glycoprotein by vanadate, shown here with pure protein, yielded full inhibition of ATPase. Thus, congruent with previous work, both sites must be intact and must interact for catalysis. Equivalent mutations (Ala or Thr) in the two catalytic sites had identical effects on a wide range of activities, emphasizing that the two catalytic sites function symmetrically. The role of the Ser-OH is to coordinate Mg(2+) in MgATP, but only at the stage of the transition state are its effects tangible. Initial substrate binding is apparently to an "open" catalytic site conformation, where the Ser-OH is dispensable. This changes to a "closed" conformation required to attain the transition state, in which the Ser-OH is a critical ligand. Formation of the latter conformation requires both sites; both sites may provide direct ligands to the transition state. PMID- 10831599 TI - Physiological non-equivalence of the two isoforms of angiotensin-converting enzyme. AB - The structurally related somatic and germinal isoforms of angiotensin-converting enzyme (ACE) contain the same catalytic active center and are encoded by the same gene, whose disruption causes renal atrophy, hypotension, and male sterility. The reason for the evolutionary conservation of both isozymes is an enigma, because, in vitro, they have very similar enzymatic properties. Despite the common enzymatic properties, discrete expression of both isoforms is maintained in alternate cell types. We have previously shown that sperm-specific expression of transgenic germinal ACE in Ace -/- male mice restores fertility without curing their other abnormalities (Ramaraj, P., Kessler, S. P., Colmenares, C. & Sen, G. C. (1998) J. Clin. Invest. 102, 371-378). In this report we tested the biological equivalence of somatic ACE and germinal ACE utilizing an in vivo isozymic substitution approach. Here we report that restoration of male fertility was not achieved by the transgenic expression of enzymatically active, somatic ACE in the sperm of Ace -/- mice. Therefore, the requisite physiological functions of the two tissue-specific isozymes of ACE are not interchangeable. PMID- 10831600 TI - Signaling lymphocytic activation molecule (CDw150) is homophilic but self associates with very low affinity. AB - Signaling lymphocytic activating molecule ((SLAM) CDw150) is a glycoprotein that belongs to the CD2 subset of the immunoglobulin superfamily and is expressed on the surface of activated T- and B-cells. It has been proposed that SLAM is homophilic and required for bidirectional signaling during T- and B-cell activation. Previous work has suggested that the affinity of SLAM self association might be unusually high, undermining the concept that protein interactions mediating transient cell-cell contacts, such as those involving leukocytes, have to be weak in order that such contacts are readily reversible. Using surface plasmon resonance-based methods and analytical ultracentrifugation (AUC), we confirm that SLAM is homophilic. However, we also establish a new theoretical treatment of surface plasmon resonance-derived homophilic binding data, which indicates that SLAM-SLAM interactions (solution K(d) approximately 200 micrometer) are in fact considerably weaker than most other well characterized protein-protein interactions at the cell surface (solution K(d) approximately 0.4-20 micrometer), a conclusion that is supported by the AUC analysis. Whereas further analysis of the AUC data imply that SLAM could form "head to head" dimers spanning adjacent cells, the very low affinity raises important questions regarding the physiological role and/or properties of such interactions. PMID- 10831601 TI - A dipeptide metalloendoprotease substrate completely blocks the response of cells in culture to cholera toxin. AB - Prior exposure (15 min at 37 degrees C) of several cell types (Vero, SH-SY5Y neuroblastoma, human intestinal epithelial T84) to 3 mm N-benzoyloxycarbonyl-Gly Phe-amide (Cbz-Gly-Phe-NH(2)), a competitive substrate for metalloendoproteases, completely suppressed cholera toxin (CT)-induced intracellular cAMP accumulation. The specificity of the inhibitory effect was demonstrated by the complete lack of effect of the dipeptide Cbz-Gly-Gly-NH(2), an inactive analogue of Cbz-Gly-Phe NH(2). The effect was reversible and dose- (IC(50) as low as 0.2 mm depending on the cell type) and time-dependent. Adding Cbz-Gly-Phe-NH(2) during the lag phase caused a diminution of its inhibitory effect similar to that observed with brefeldin A (BFA). Whereas the dipeptide completely suppressed the CT-induced adenylate cyclase (AC) activity, a direct effect on AC is unlikely since the elevation of intracellular cAMP by forskolin was only slightly reduced. The A(1) peptide of CT and NAD(+) activated the AC to the same extent in membranes from control and Cbz-Gly-Phe-NH(2)-treated cells or when Cbz-Gly-Phe-NH(2) was added directly to the assay. The inhibitory effects of suboptimal amounts of Cbz-Gly Phe-NH(2) and BFA were not additive pointing to a similar mode of action of the two substances. However, Madin-Darby canine kidney cells of which the Golgi structure is BFA-resistant were not resistant to the inhibitory action of Cbz-Gly Phe-NH(2) on CT cytotoxicity. Several lines of evidence indicate that a perturbation of intracellular Ca(2+) homeostasis by Cbz-Gly-Phe-NH(2) is not responsible for the inhibitory effect of the dipeptide. The dipeptide had also no effect on the binding of (125)I-CT to cells and even increased its intracellular internalization. In contrast with BFA, Cbz-Gly-Phe-NH(2) did not completely suppress the formation of the catalytically active A(1) fragment from bound CT. The data are compatible with a role of metalloendoprotease activity in the intracellular trafficking and processing of CT, although other mechanisms of action of Cbz-Gly-Phe-NH(2) cannot be excluded. PMID- 10831602 TI - Syndecan-regulated receptor signaling. PMID- 10831603 TI - It's got you covered. NF-kappaB in the epidermis. PMID- 10831604 TI - JIL-1, a chromosomal kinase implicated in regulation of chromatin structure, associates with the male specific lethal (MSL) dosage compensation complex. AB - JIL-1 is a novel chromosomal kinase that is upregulated almost twofold on the male X chromosome in Drosophila. Here we demonstrate that JIL-1 colocalizes and physically interacts with male specific lethal (MSL) dosage compensation complex proteins. Furthermore, ectopic expression of the MSL complex directed by MSL2 in females causes a concomitant upregulation of JIL-1 to the female X that is abolished in msl mutants unable to assemble the complex. Thus, these results strongly indicate JIL-1 associates with the MSL complex and further suggests JIL 1 functions in signal transduction pathways regulating chromatin structure. PMID- 10831605 TI - Inhibition of apoptotic signaling cascades causes loss of trophic factor dependence during neuronal maturation. AB - During development, neurons are acutely dependent on target-derived trophic factors for survival. This dependence on trophic support decreases dramatically with maturation in several neuronal populations, including sympathetic neurons. Analyses of nerve growth factor deprivation in immature and mature sympathetic neurons indicate that maturation aborts the cell death pathway at a point that is mechanistically indistinguishable from Bax deletion. However, neither the mRNA nor protein level of BAX changes with neuronal maturation. Therefore, BAX must be regulated posttranslationally in mature neurons. Nerve growth factor deprivation in immature sympathetic neurons induces two parallel processes: (a) a protein synthesis-dependent, caspase-independent translocation of BAX from the cytosol to mitochondria, followed by mitochondrial membrane integration and loss of cytochrome c; and (b) the development of competence-to-die, which requires neither macromolecular synthesis nor BAX expression. Activation of both signaling pathways is required for caspase activation and apoptosis in immature sympathetic neurons. In contrast, nerve growth factor withdrawal in mature sympathetic neurons did not induce the translocation of either BAX or cytochrome c. Moreover, mature neurons did not develop competence-to-die with cytoplasmic accumulation of cytochrome c. Therefore, inhibition of both BAX-dependent cytochrome c release and the development of competence-to-die contributed to the loss of trophic factor dependence associated with neuronal maturation. PMID- 10831606 TI - Leydig cell loss and spermatogenic arrest in platelet-derived growth factor (PDGF)-A-deficient mice. AB - Platelet-derived growth factor (PDGF)- A-deficient male mice were found to develop progressive reduction of testicular size, Leydig cells loss, and spermatogenic arrest. In normal mice, the PDGF-A and PDGF-Ralpha expression pattern showed positive cells in the seminiferous epithelium and in interstitial mesenchymal cells, respectively. The testicular defects seen in PDGF-A-/- mice, combined with the normal developmental expression of PDGF-A and PDGF-Ralpha, indicate that through an epithelial-mesenchymal signaling, the PDGF-A gene is essential for the development of the Leydig cell lineage. These findings suggest that PDGF-A may play a role in the cascade of genes involved in male gonad differentiation. The Leydig cell loss and the spermatogenic impairment in the mutant mice are reminiscent of cases of testicular failure in man. PMID- 10831607 TI - Yeast nucleoporins involved in passive nuclear envelope permeability. AB - The vertebrate nuclear pore complex (NPC) harbors an approximately 10-nm diameter diffusion channel that is large enough to admit 50-kD polypeptides. We have analyzed the permeability properties of the Saccharomyces cerevisiae nuclear envelope (NE) using import (NLS) and export (NES) signal-containing green fluorescent protein (GFP) reporters. Compared with wild-type, passive export rates of a classical karyopherin/importin (Kap) Kap60p/Kap95p-targeted NLS-GFP reporter (cNLS-GFP) were significantly faster in nup188-Delta and nup170-Delta cells. Similar results were obtained using two other NLS-GFP reporters, containing either the Kap104p-targeted Nab2p NLS (rgNLS) or the Kap121p-targeted Pho4p NLS (pNLS). Elevated levels of Hsp70 stimulated cNLS-GFP import, but had no effect on the import of rgNLS-GFP. Thus, the role of Hsp70 in NLS-directed import may be NLS- or targeting pathway-specific. Equilibrium sieving limits for the diffusion channel were assessed in vivo using NES-GFP reporters of 36-126 kD and were found to be greater than wild-type in nup188-Delta and nup170-Delta cells. We propose that Nup170p and Nup188p are involved in establishing the functional resting diameter of the NPC's central transport channel. PMID- 10831608 TI - PNG1, a yeast gene encoding a highly conserved peptide:N-glycanase. AB - It has been proposed that cytoplasmic peptide:N-glycanase (PNGase) may be involved in the proteasome-dependent quality control machinery used to degrade newly synthesized glycoproteins that do not correctly fold in the ER. However, a lack of information about the structure of the enzyme has limited our ability to obtain insight into its precise biological function. A PNGase-defective mutant (png1-1) was identified by screening a collection of mutagenized strains for the absence of PNGase activity in cell extracts. The PNG1 gene was mapped to the left arm of chromosome XVI by genetic approaches and its open reading frame was identified. PNG1 encodes a soluble protein that, when expressed in Escherichia coli, exhibited PNGase activity. PNG1 may be required for efficient proteasome mediated degradation of a misfolded glycoprotein. Subcellular localization studies indicate that Png1p is present in the nucleus as well as the cytosol. Sequencing of expressed sequence tag clones revealed that Png1p is highly conserved in a wide variety of eukaryotes including mammals, suggesting that the enzyme has an important function. PMID- 10831609 TI - The role of intraorganellar Ca(2+) in late endosome-lysosome heterotypic fusion and in the reformation of lysosomes from hybrid organelles. AB - We have investigated the requirement for Ca(2+) in the fusion and content mixing of rat hepatocyte late endosomes and lysosomes in a cell-free system. Fusion to form hybrid organelles was inhibited by 1,2-bis(2-aminophenoxy) ethane-N,N,N',N' tetraacetic acid (BAPTA), but not by EGTA, and this inhibition was reversed by adding additional Ca(2+). Fusion was also inhibited by methyl ester of EGTA (EGTA AM), a membrane permeable, hydrolyzable ester of EGTA, and pretreatment of organelles with EGTA-AM showed that the chelation of lumenal Ca(2+) reduced the amount of fusion. The requirement for Ca(2+) for fusion was a later event than the requirement for a rab protein since the system became resistant to inhibition by GDP dissociation inhibitor at earlier times than it became resistant to BAPTA. We have developed a cell-free assay to study the reformation of lysosomes from late endosome-lysosome hybrid organelles that were isolated from the rat liver. The recovery of electron dense lysosomes was shown to require ATP and was inhibited by bafilomycin and EGTA-AM. The data support a model in which endocytosed Ca(2+) plays a role in the fusion of late endosomes and lysosomes, the reformation of lysosomes, and the dynamic equilibrium of organelles in the late endocytic pathway. PMID- 10831610 TI - SNAREpins are functionally resistant to disruption by NSF and alphaSNAP. AB - SNARE (SNAP [soluble NSF (N-ethylmaleimide-sensitive fusion protein) attachment protein] receptor) proteins are required for many fusion processes, and recent studies of isolated SNARE proteins reveal that they are inherently capable of fusing lipid bilayers. Cis-SNARE complexes (formed when vesicle SNAREs [v-SNAREs] and target membrane SNAREs [t-SNAREs] combine in the same membrane) are disrupted by the action of the abundant cytoplasmic ATPase NSF, which is necessary to maintain a supply of uncombined v- and t-SNAREs for fusion in cells. Fusion is mediated by these same SNARE proteins, forming trans-SNARE complexes between membranes. This raises an important question: why doesn't NSF disrupt these SNARE complexes as well, preventing fusion from occurring at all? Here, we report several lines of evidence that demonstrate that SNAREpins (trans-SNARE complexes) are in fact functionally resistant to NSF, and they become so at the moment they form and commit to fusion. This elegant design allows fusion to proceed locally in the face of an overall environment that massively favors SNARE disruption. PMID- 10831611 TI - Zyxin, a regulator of actin filament assembly, targets the mitotic apparatus by interacting with h-warts/LATS1 tumor suppressor. AB - The mitotic apparatus plays a pivotal role in dividing cells to ensure each daughter cell receives a full set of chromosomes and complement of cytoplasm during mitosis. A human homologue of the Drosophila warts tumor suppressor, h warts/LATS1, is an evolutionarily conserved serine/threonine kinase and a dynamic component of the mitotic apparatus. We have identified an interaction of h warts/LATS1 with zyxin, a regulator of actin filament assembly. Zyxin is a component of focal adhesion, however, during mitosis a fraction of cytoplasmic dispersed zyxin becomes associated with h-warts/LATS1 on the mitotic apparatus. We found that zyxin is phosphorylated specifically during mitosis, most likely by Cdc2 kinase, and that the phosphorylation regulates association with h warts/LATS1. Furthermore, microinjection of truncated h-warts/LATS1 protein, including the zyxin-binding portion, interfered with localization of zyxin to mitotic apparatus, and the duration of mitosis of these injected cells was significantly longer than that of control cells. These findings suggest that h warts/LATS1 and zyxin play a crucial role in controlling mitosis progression by forming a regulatory complex on mitotic apparatus. PMID- 10831612 TI - ADP ribosylation factor-like protein 2 (Arl2) regulates the interaction of tubulin-folding cofactor D with native tubulin. AB - The ADP ribosylation factor-like proteins (Arls) are a family of small monomeric G proteins of unknown function. Here, we show that Arl2 interacts with the tubulin-specific chaperone protein known as cofactor D. Cofactors C, D, and E assemble the alpha/beta- tubulin heterodimer and also interact with native tubulin, stimulating it to hydrolyze GTP and thus acting together as a beta tubulin GTPase activating protein (GAP). We find that Arl2 downregulates the tubulin GAP activity of C, D, and E, and inhibits the binding of D to native tubulin in vitro. We also find that overexpression of cofactors D or E in cultured cells results in the destruction of the tubulin heterodimer and of microtubules. Arl2 specifically prevents destruction of tubulin and microtubules by cofactor D, but not by cofactor E. We generated mutant forms of Arl2 based on the known properties of classical Ras-family mutations. Experiments using these altered forms of Arl2 in vitro and in vivo demonstrate that it is GDP-bound Arl2 that interacts with cofactor D, thereby averting tubulin and microtubule destruction. These data establish a role for Arl2 in modulating the interaction of tubulin-folding cofactors with native tubulin in vivo. PMID- 10831613 TI - Polyglycylation of tubulin is essential and affects cell motility and division in Tetrahymena thermophila. AB - We analyzed the role of tubulin polyglycylation in Tetrahymena thermophila using in vivo mutagenesis and immunochemical analysis with modification-specific antibodies. Three and five polyglycylation sites were identified at glutamic acids near the COOH termini of alpha- and beta-tubulin, respectively. Mutants lacking all polyglycylation sites on alpha-tubulin have normal phenotype, whereas similar sites on beta-tubulin are essential. A viable mutant with three mutated sites in beta-tubulin showed reduced tubulin glycylation, slow growth and motility, and defects in cytokinesis. Cells in which all five polyglycylation sites on beta-tubulin were mutated were viable if they were cotransformed with an alpha-tubulin gene whose COOH terminus was replaced by the wild-type COOH terminus of beta-tubulin. In this double mutant, beta-tubulin lacked detectable polyglycylation, while the alpha-beta tubulin chimera was hyperglycylated compared with alpha-tubulin in wild-type cells. Thus, the essential function of polyglycylation of the COOH terminus of beta-tubulin can be transferred to alpha tubulin, indicating it is the total amount of polyglycylation on both alpha- and beta-tubulin that is essential for survival. PMID- 10831614 TI - Ges, A human GTPase of the Rad/Gem/Kir family, promotes endothelial cell sprouting and cytoskeleton reorganization. AB - Rad, Gem/Kir, and mRem (RGK) represent a unique GTPase family with largely unknown functions (Reynet, C., and C.R. Kahn. 1993. Science. 262:1441-1444; Cohen, L., R. Mohr, Y. Chen, M. Huang, R. Kato, D. Dorin, F. Tamanoi, A. Goga, D. Afar, N. Rosenberg, and O. Witte. Proc. Natl. Acad. Sci. USA. 1994. 91:12448 12452; Maguire, J., T. Santoro, P. Jensen, U. Siebenlist, J. Yewdell, and K. Kelly. 1994. Science. 265:241-244; Finlin, B.S., and D.A. Andres. 1997. J. Biol. Chem. 272:21982-21988). We report that Ges (GTPase regulating endothelial cell sprouting), a human RGK protein expressed in the endothelium, functions as a potent morphogenic switch in endothelial cells (ECs). Ges function is sufficient to substitute for angiogenic growth factor/extracellular matrix (ECM) signals in promoting EC sprouting, since overexpression of Ges in ECs cultured on glass leads to the development of long cytoplasmic extensions and reorganization of the actin cytoskeleton. Ges function is also necessary for Matrigel-induced EC sprouting, since this event is blocked by its dominant negative mutant, Ges(T94N), predicted to prevent the activation of endogenous Ges through sequestration of its guanine nucleotide exchange factor. Thus, Ges appears to be a key transducer linking extracellular signals to cytoskeleton/morphology changes in ECs. PMID- 10831615 TI - Downregulation of 14-3-3sigma prevents clonal evolution and leads to immortalization of primary human keratinocytes. AB - In human epidermal keratinocytes, replicative senescence, is determined by a progressive decline of clonogenic and dividing cells. Its timing is controlled by clonal evolution, that is, by the continuous transition from stem cells to transient amplifying cells. We now report that downregulation of 14-3-3sigma, which is specifically expressed in human stratified epithelia, prevents keratinocyte clonal evolution, thereby forcing keratinocytes into the stem cell compartment. This allows primary human keratinocytes to readily escape replicative senescence. 14-3-3sigma-dependent bypass of senescence is accompanied by maintenance of telomerase activity and by downregulation of the p16(INK4a) tumor suppressor gene, hallmarks of keratinocyte immortalization. Taken together, these data therefore suggest that inhibition of a single endogenous gene product fosters immortalization of primary human epithelial cells without the need of exogenous oncogenes and/or oncoviruses. PMID- 10831616 TI - Observing FcepsilonRI signaling from the inside of the mast cell membrane. AB - We have determined the membrane topography of the high-affinity IgE receptor, FcstraightepsilonRI, and its associated tyrosine kinases, Lyn and Syk, by immunogold labeling and transmission electron microscopic (TEM) analysis of membrane sheets prepared from RBL-2H3 mast cells. The method of Sanan and Anderson (Sanan, D.A., and R.G.W. Anderson. 1991. J. Histochem. Cytochem. 39:1017 1024) was modified to generate membrane sheets from the dorsal surface of RBL-2H3 cells. Signaling molecules were localized on the cytoplasmic face of these native membranes by immunogold labeling and high-resolution TEM analysis. In unstimulated cells, the majority of gold particles marking both FcepsilonRI and Lyn are distributed as small clusters (2-9 gold particles) that do not associate with clathrin-coated membrane. Approximately 25% of FcepsilonRI clusters contain Lyn. In contrast, there is essentially no FcepsilonRI-Syk colocalization in resting cells. 2 min after FcepsilonRI cross-linking, approximately 10% of Lyn colocalizes with small and medium-sized FcepsilonRI clusters (up to 20 gold particles), whereas approximately 16% of Lyn is found in distinctive strings and clusters at the periphery of large receptor clusters (20-100 gold particles) that form on characteristically osmiophilic membrane patches. While Lyn is excluded, Syk is dramatically recruited into these larger aggregates. The clathrin-coated pits that internalize cross-linked receptors bud from membrane adjacent to the Syk-containing receptor complexes. The sequential association of FcstraightepsilonRI with Lyn, Syk, and coated pits in topographically distinct membrane domains implicates membrane segregation in the regulation of FcstraightepsilonRI signaling. PMID- 10831617 TI - The cysteine-rich domain of human ADAM 12 supports cell adhesion through syndecans and triggers signaling events that lead to beta1 integrin-dependent cell spreading. AB - The ADAMs (a disintegrin and metalloprotease) family of proteins is involved in a variety of cellular interactions, including cell adhesion and ecto- domain shedding. Here we show that ADAM 12 binds to cell surface syndecans. Three forms of recombinant ADAM 12 were used in these experiments: the cys-teine-rich domain made in Escherichia coli (rADAM 12-cys), the disintegrin-like and cysteine-rich domain made in insect cells (rADAM 12-DC), and full-length human ADAM 12-S tagged with green fluorescent protein made in mammalian cells (rADAM 12-GFP). Mesenchymal cells specifically and in a dose-dependent manner attach to ADAM 12 via members of the syndecan family. After binding to syndecans, mesenchymal cells spread and form focal adhesions and actin stress fibers. Integrin beta1 was responsible for cell spreading because function-blocking monoclonal antibodies completely inhibited cell spreading, and chondroblasts lacking beta1 integrin attached but did not spread. These data suggest that mesenchymal cells use syndecans as the initial receptor for the ADAM 12 cysteine-rich domain-mediated cell adhesion, and then the beta1 integrin to induce cell spreading. Interestingly, carcinoma cells attached but did not spread on ADAM 12. However, spreading could be efficiently induced by the addition of either 1 mM Mn(2+) or the beta1 integrin-activating monoclonal antibody 12G10, suggesting that in these carcinoma cells, the ADAM 12-syndecan complex fails to modulate the function of beta1 integrin. PMID- 10831619 TI - Transepithelial chloride secretion and cystogenesis in autosomal dominant polycystic kidney disease. PMID- 10831618 TI - Tissue plasminogen activator-mediated fibrinolysis protects against axonal degeneration and demyelination after sciatic nerve injury. AB - Tissue plasminogen activator (tPA) is a serine protease that converts plasminogen to plasmin and can trigger the degradation of extracellular matrix proteins. In the nervous system, under noninflammatory conditions, tPA contributes to excitotoxic neuronal death, probably through degradation of laminin. To evaluate the contribution of extracellular proteolysis in inflammatory neuronal degeneration, we performed sciatic nerve injury in mice. Proteolytic activity was increased in the nerve after injury, and this activity was primarily because of Schwann cell-produced tPA. To identify whether tPA release after nerve damage played a beneficial or deleterious role, we crushed the sciatic nerve of mice deficient for tPA. Axonal demyelination was exacerbated in the absence of tPA or plasminogen, indicating that tPA has a protective role in nerve injury, and that this protective effect is due to its proteolytic action on plasminogen. Axonal damage was correlated with increased fibrin(ogen) deposition, suggesting that this protein might play a role in neuronal injury. Consistent with this idea, the increased axonal degeneration phenotype in tPA- or plasminogen-deficient mice was ameliorated by genetic or pharmacological depletion of fibrinogen, identifying fibrin as the plasmin substrate in the nervous system under inflammatory axonal damage. This study shows that fibrin deposition exacerbates axonal injury, and that induction of an extracellular proteolytic cascade is a beneficial response of the tissue to remove fibrin. tPA/plasmin-mediated fibrinolysis may be a widespread protective mechanism in neuroinflammatory pathologies. PMID- 10831620 TI - Renal failure and hantavirus infection in Europe. PMID- 10831621 TI - Human polyomavirus BKV and renal disease. PMID- 10831622 TI - Sympathetic overactivity in renal failure controlled by ACE inhibition: clinical significance. PMID- 10831623 TI - The role of the cardiac endothelin system in heart failure. PMID- 10831624 TI - Why and how to monitor bacterial contamination of dialysate? PMID- 10831625 TI - Atrial fibrillation in dialysis patients. PMID- 10831626 TI - Nephrology services in Pakistan. PMID- 10831627 TI - Streptococcal M protein enhances TGF-beta production and increases surface IgA positive B cells in vitro in IgA nephropathy. AB - BACKGROUND: High serum levels and enhanced in vitro production of IgA are observed in more than half of patients with IgA nephropathy (IgAN); and transforming forming growth factor-beta (TGF-beta) is certain IgA class switching factor. On the other hand, macroscopic haematuria appears frequently with upper respiratory infection as tonsillitis in IgAN. METHODS: We compared the lymphocytic response to in-vitro stimulation by group A streptococcal M proteins of apparent virulence factor between IgAN, non-proliferative glomerulonephritis (NPGN), and normal subjects. M proteins were extracted from group A streptococcal strain type 5 and type 12 determined serologically. RESULTS: M protein-induced proliferation of lymphocytes from IgAN was higher than in NPGN but not in healthy control subjects. Flow cytometric analysis indicated that stimulation by M protein extracts derived from type 5 streptococci (M5) increased surface IgA positive B cells in IgAN, but did not activate the production of soluble IgA. We also showed that M5 induced significant increases in TGF-beta, in culture supernatants of lymphocytes from patients with IgAN. CONCLUSION: Our results suggest that Streptococcal infection may play an important role in the pathogenesis of IgAN by stimulating IgA production through TGF-beta synthesis. PMID- 10831628 TI - Characterization of a DNA binding site that mediates the stimulatory effect of cyclosporin-A on type III collagen expression in renal cells. AB - BACKGROUND: Previous work from our laboratory demonstrated upregulation of type III collagen by cyclosporin A (CsA) in a cellular model of renal fibroblasts 'in vitro', suggesting that a mechanism of gene transcriptional activation might be responsible for collagen accumulation in renal fibrosis resulting from chronic CsA treatment. METHODS: We analysed in the same cellular model: (i) COL3A1 mRNA expression by RT-PCR; (ii) COL3A1 promoter activity by transfection of renal fibroblasts with constructs containing promoter fragments of different length fused to a reporter gene; (iii) expression of transcription factors by western blot analysis; (iv) DNA-protein binding by gel retardation assays with nuclear extracts from CsA-treated and untreated cells; and (v) site-directed mutagenesis of COL3A1 promoter to verify the role of a short DNA segment as CsA responsive element. RESULTS: CsA induced a 3-5-fold increase in COL3A1 mRNA that was paralleled by a stimulation of the COL3A1 promoter. Degradation of COL3A1 mRNA was comparable in CsA-treated and -untreated cells. The target region was first limited to a 178 bp fragment from -117 to +61 (pFV1). By gel retardation, utilizing several oligonucleotides that covered the whole length of pFV1, we detected a factor able to bind the promoter DNA (oligo 31) in nuclear extracts after 3 h treatment with CsA. The binding was absent in untreated cells and it was not detected when a 10-base mutation was introduced in oligonucleotide 31. Finally, the same substitution mutation at the site of binding of this factor abolished the stimulatory effect of CsA on COL3A1 promoter. Some transcription factors, whose potential binding sites are included in the above promoter fragment, were induced by CsA treatment either soon (3 h) or late (24-72 h) after treatment and were detected by western blot analysis. CONCLUSIONS: CsA induces the synthesis of type III collagen by stimulating a pathway leading to activation of COL3A1 promoter and upregulation of COL3A1 mRNA. A short promoter fragment, proximal to the transcription start site, is the target of CsA stimulation. PMID- 10831629 TI - Effects of growth hormone on renal renin gene expression in normal rats and rats with myocardial infarction. AB - BACKGROUND: Published data regarding effects of growth hormone (GH) on the renin system are controversial. The aim of this study therefore was to evaluate the effects of GH on the renin system in normal rats and rats with myocardial infarction (MI). METHODS: Normal rats received 2, 5, or 10 IU GH/kg/day or vehicle subcutaneously for 4 weeks. Furthermore rats with MI were randomized to receive 2 IU GH/kg/day or vehicle for 4 weeks. Subdivision into MI groups (mild, moderate, and large) was by histological determination of infarct size. Renal renin gene expression was assessed by RNAase protection assay and plasma renin activity by radioimmunoassay. In addition, isolated mouse juxtaglomerular cells were exposed to GH for 20 h, and renin secretion rates were assessed. RESULTS: GH treatment in normal rats for 4 weeks increased body weight, and kidney weight to body weight ratio, but did not affect renin secretion and renal renin gene expression. In rats with large MI, renal renin gene expression increased about fourfold, but was unchanged in rats with small and moderate MI as compared to normal rats. In rats with MI, body weight decreased and this decrease was partially reversed by GH treatment. GH treatment did not change renal renin gene expression, and renin secretion in rats with MI. Renin secretion of isolated juxtaglomerular cells was unaffected by GH. CONCLUSIONS: Our study demonstrates that GH treatment has no significant effect on renin secretion and on renal renin gene expression in normal rats and in rats with stimulated renin system due to MI in vivo. In isolated juxtaglomerular cells in vitro, renin secretion was also unaffected by GH. PMID- 10831630 TI - L-type calcium current of isolated rat cardiac myocytes in experimental uraemia. AB - BACKGROUND: End-stage renal failure is associated with a low-output cardiomyopathy, left ventricular hypertrophy and increased QTc dispersion. Cardiac dysfunction is prevalent in patients at the beginning of dialysis and is an important predictor of mortality. Ca(2+) influx through voltage-gated L-type Ca(2+) channels plays a key role in the excitation-contraction coupling of cardiac myocytes. The purpose of this study was to examine the effect of subtotal nephrectomy (SNx) in the rat on both cardiac L-type Ca(2+) currents and action potential duration. METHODS: Wistar rats underwent two-stage SNx; control rats (C) underwent bilateral renal decapsulation. Animals were sacrificed after 8 weeks, and ventricular myocytes were isolated. SNx rats showed a 2-fold increase in plasma urea and creatinine compared with C rats. Whole-cell patch clamp techniques were used to examine L-type Ca(2+) channel currents in isolated cardiac myocytes at 37 degrees C. In separate experiments, the epicardial monophasic action potentials of isolated perfused whole hearts from C and SNx rats were recorded. RESULTS: The amplitude and current-voltage relationships of the L-type Ca(2+) current were not significantly different in myocytes from C (n=11) and SNx (n=8) rats. However, the rate of inactivation of the Ca(2+) current was increased by approximately 15-25% (P<0. 05) in myocytes from SNx rats. The action potential duration (APD(33)) at the apex of the left ventricle was approximately 20% shorter (P<0.01) in hearts from SNx rats as compared with controls. CONCLUSIONS: Renal failure is associated with rapid inactivation of cardiac ventricular myocyte L-type Ca(2+) currents, which may reduce Ca(2+) influx and contribute to shortening of the action potential duration. PMID- 10831631 TI - Severe proteinuria, sustained for 6 months, induces tubular epithelial cell injury and cell infiltration in rats but not progressive interstitial fibrosis. AB - BACKGROUND: Sustained proteinuria is reported to be very harmful to the tubulointerstitium, leading to severe interstitial injury. However, it remains unclear whether sustained proteinuria itself is responsible for severe interstitial injury because, in the previously reported models, the development of factors other than proteinuria in tubulointerstitial lesions could not be excluded completely. METHODS: After treatment to induce immune tolerance to mouse immunoglobulin, 20 rats were injected with anti-rat slit diaphragm monoclonal antibody (mAb) 5-1-6 twice a week for 6 months and were then sacrificed. RESULTS: mAb 5-1-6 induced massive proteinuria in 11 rats. In nine rats with mild proteinuria, no histological alteration could be detected with light microscopy and immunofluorescence. In nephrotic rats, light microscopy showed minor glomerular abnormalities, with interstitial oedema, tubular epithelial cell degeneration and interstitial cell infiltration. Immunofluorescence revealed increased expression of vimentin and an increased number of OX1-, OX19- and ED1 positive cells. However, we could not detect any accumulation of type I and IV collagen or laminin in the tubulointerstitium. RT-PCR showed that the expression of mRNA for type I collagen was not increased, compared with that in control rats. CONCLUSIONS: We succeeded in developing a model of persistent nephrosis without severe glomerular abnormalities, nephrectomy or other manoeuvres known to induce disturbed haemodynamics, using an agent without tubulointerstitial toxicity, and considered it to be suitable for investigating the direct toxicity of proteinuria. In this model, isolated massive proteinuria induced interstitial injury. However, the degree of injury was suggested to be much less than that observed in other previously developed models. PMID- 10831632 TI - Aminoguanidine induces haematuria of non-glomerular origin in spontaneously hypertensive rats. AB - BACKGROUND: Administration of N(G)-nitro-L-arginine methyl ester (L-NAME), a non selective inhibitor of nitric oxide synthase (NOS), induces glomerulosclerosis in spontaneously hypertensive rats (SHR). We investigated the effects of administering aminoguanidine (AG), a selective inhibitor of inducible NOS (iNOS), on glomerular histology, serum creatinine concentration, albuminuria and haematuria in SHR. METHODS: SHR and Wistar Kyoto rats (WKR) (age, 7 weeks) were given a daily water supply with or without 0.1% AG. Every 4 weeks, 24 h urine samples were collected and checked for haematuria by a dipstick method, and systolic blood pressure was measured. After 16 weeks, serum creatinine, albuminuria and glomerulosclerosis indices (GSI) were evaluated, and the size of urinary erythrocytes in AG-treated SHR was measured by flow cytometry. Glomeruli were observed by transmission and scanning electron microscopy. Some AG-treated SHR received a furosemide injection and then urinary erythrocyte size was determined. RESULTS: Systolic blood pressure, serum creatinine, albuminuria and GSI were similar between the untreated and AG-treated groups in both strains. However, AG treatment induced significant haematuria in SHR, but not in WKR. Electron microscopy did not provide any evidence for glomerular bleeding sites in AG-treated SHR. In urine with osmolalities exceeding 750 mOsm/kg, haematuria of AG-treated SHR consisted of erythrocytes smaller in size than venous erythrocytes. After furosemide injection leading to near isotonic urine, the size of urinary erythrocytes was similar to that of venous erythrocytes. CONCLUSIONS: The absence of morphological evidence for glomerular bleeding sites and similar intrinsic size between urinary and venous erythrocytes suggest that AG induces a non-glomerular type of haematuria in SHR. PMID- 10831633 TI - Evidence of further genetic heterogeneity in autosomal dominant medullary cystic kidney disease. AB - BACKGROUND: Autosomal dominant medullary cystic kidney disease is a genetically heterogeneous nephropathy with clinical and morphological features similar to recessively inherited juvenile nephronophthisis. Recently, a second gene locus on chromosome 16p12, MCKD2 has been mapped [1] in addition to the known locus on chromosome 1q21 (MCKD1) [2]. In a previous study we have excluded linkage for three caucasian families to the MCKD1 locus [3]. METHODS: Haplotype analysis was performed on 72 individuals (including 24 affected subjects), using a set of seven microsatellite markers spanning the critical region on chromosome 16p12-p13 of about 10.5 cM. RESULTS: We report on haplotype analysis of closely linked markers to the MCKD2 locus in the previously studied families and two additional families. CONCLUSION: In all five families the association of MCKD2 with the disease was excluded by a multipoint LOD score <-2, thus suggesting the involvement of a third MCKD locus. PMID- 10831634 TI - Aminoglycosides and renal magnesium homeostasis in humans. AB - BACKGROUND: The use of aminoglycosides has been linked with hypomagnesaemia in scattered reports. The objective of the study was to measure prospectively the effect of treatment with the aminoglycoside amikacin on renal magnesium homeostasis. METHODS: Twenty-four cystic fibrosis patients (aged 9-19 years) admitted because of exacerbation of pulmonary symptoms caused by Pseudomonas aeruginosa were treated with the aminoglycoside amikacin and the cephalosporin ceftazidime for 14 days. Renal values and plasma and urinary electrolytes were measured before and at the end of the systemic anti-pseudomonal therapy. RESULTS: In the patients with cystic fibrosis, treatment with amikacin and ceftazidime did not modify plasma creatinine or urea and plasma or urinary sodium, potassium and calcium. Treatment with amikacin and ceftazidime significantly decreased both plasma total magnesium (from 0.77 (0. 74-0.81) to 0.73 (0.71-75) mmol/l; median and interquartile range) and ionized magnesium (from 0.53 (0.50-0.55) to 0.50 (0.47-0.52) mmol/l) concentration and increased fractional urinary magnesium excretion (from 0.0568 (0.0494-0.0716) to 0.0721 (0.0630-0.111)) and total urinary magnesium excretion (from 30.7 (26.5-38.0) to 38.5 (31. 5-49.0) micromol/l glomerular filtration rate). CONCLUSIONS: The present study demonstrates that systemic therapy with amikacin plus ceftazidime causes mild hypomagnesaemia secondary to renal magnesium wasting even in the absence of a significant rise in circulating creatinine and urea. PMID- 10831635 TI - Duplex sonographic registration of age and diabetes-related loss of renal vasodilatory response to nitroglycerine. AB - BACKGROUND: Nitroglycerine effects dilatation of the vas afferens in the kidney by its active metabolite nitric oxide (NO). In diabetic nephropathy, NO appears to cause hyperfiltration and proteinuria. The aim of this study was to investigate the potential of duplex Doppler ultrasonography in recording changes in resistive indices following nitroglycerine and whether the extent of changes in diabetic nephropathy is reduced as a result of the arteriolar dilatation mediated by NO. METHODS: Fifty-three subjects made up three groups: group 1, 12 young healthy volunteers (5 male, 7 female, 28+/-6 years); group 2, 21 older healthy volunteers (7 male, 14 female, 62+/-8 years); group 3, 20 patients (13 male, 7 female, 56+/-18 years) with mild diabetic nephropathy. The resistive index (RI) was measured in the interlobar arteries before, and 1, 3 and 5 min after administration of 0.8 mg sublingual nitroglycerine. RESULTS: The initial RI of 0.592 decreased in group 1 by 10.8% (P<0. 01) after nitroglycerine, in group 2 the initial figure of 0.631 decreased by 5.9% (P<0.01), and in group 3 the initial figure of 0. 669 decreased by 3.4% (P<0.01). Initial RI values differed significantly between groups 1 and 3 (P<0.01) and groups 1 and 2 (P<0.01), as did values between all healthy volunteers (groups 1 and 2, n=33) and patients with nephropathy (P<0.005). The extent of DeltaRI differed significantly between groups 1 and 3 (P<0.01), 1 and 2 (P<0.02), and between all volunteers and group 3 (P<0.003). In groups 1 and 2 the initial RI was dependent on age (P<0.03), in group 3 on age and creatinine clearance (P<0.02 and P<0.05 respectively). DeltaRI correlated with age in the healthy subjects (P<0.01) and with duration of diabetes in diabetic nephropathy (P0. 03). CONCLUSIONS: Haemodynamic changes in renal perfusion caused by nitroglycerine can be detected using duplex ultrasonography. In healthy subjects, DeltaRI declines with increasing age. In diabetic nephropathy patients, the response to nitroglycerine is reduced. The increased initial RI in such patients suggests fixed arteriolar damage. PMID- 10831636 TI - Steroid and cyclophosphamide in IgA nephropathy. AB - BACKGROUND: IgA nephropathy is associated with a wide spectrum of possible lesions. Therefore, different responses to anti-inflammatory or immunosuppressive therapies should be expected with acute inflammatory changes, which are predominantly reversible, and with prevalently sclerotic lesions. METHODS: The effects of a combined schedule of prednisone and cyclophosphamide was analysed in the specific subset of IgA nephropathy patients with acute inflammatory histologic changes associated with haematuria and proteinuria. Two groups of patients, with similar histologic lesions and clinical presentation, were considered. The first group (12 patients) was treated within 1 week after renal biopsy; starting with three pulses of methylprednisolone (1 g) followed by oral prednisone (0.8 mg/kg body weight for 2 weeks, 0.6 mg/kg for another 2 weeks, 0.4 mg/kg for an additional 4 weeks, then slowly tapered by 5 mg each month until discontinuation) and 1.5 mg/kg cyclophosphamide for 2 months. A second sample of eight untreated patients served as a control group. Treated and untreated patients had diffuse mesangial proliferation with florid crescents (8-60% in treated and 10-40% in untreated patients) with mild degree of glomerular sclerosis and interstitial changes. Basal creatinine (167 micromol/l, range 79 371 vs 132 micromol/l, range 79-256) and proteinuria (3.0 g/24 h, 1.0-4.9 vs 3.3 g/24 h, 1.0-13.7) were not statistically different between treated and untreated patients respectively. Nine treated and six untreated patients were hypertensive. Blood pressure treatment did not include ACE-inhibitors. RESULTS: Untreated patients' 5-year renal survival, as assessed by the Kaplan-Meier method, was found to be significantly lower than treated patients (37.5 vs 91.6%, log-rank P=0.01 and Breslow test P=0.008; relative risk to reach the endpoint of a 100% increase in serum creatinine=3.58, P=0.03). CONCLUSION: This short course of therapy with prednisone and cyclophosphamide has been effective in a subset of IgA nephropathy patients with florid glomerular changes and major urinary abnormalities, turning off phlogistic activity and preventing subsequent progression toward renal failure. PMID- 10831637 TI - The ACE insertion/deletion polymorphism has no influence on progression of renal function loss in autosomal dominant polycystic kidney disease. AB - BACKGROUND: Autosomal dominant polycystic kidney disease (ADPKD) shows a variable clinical course that is not fully explained by the genetic heterogeneity of this disease. We looked for a possible genetic modifier, the ACE I/D polymorphism, and its influence on progression towards end-stage renal failure (ESRF). METHODS: Forty-nine ADPKD patients who reached ESRF <40 years, and 21 PKD1 patients who reached ESRF > 60 years or were not on dialysis at 60 years of age were recruited. Clinical data were provided by questionnaires. Blood was collected for the determination of the ACE insertion/deletion (I/D) polymorphism genotype. The ACE genotype was also determined in a general, control PKD1 group (n=59). RESULTS: Patients who reached ESRF <40 years had significantly more early onset hypertension than patients reaching ESRF >60 years (80% vs 21%; P<0.001). The ACE genotype distribution showed no differences between the groups of the rapid progressors (DD 20%, ID 56%, II 24%), the slow progressors (DD 29%, ID 52%, II 19%) and the general PKD1 control population (DD 31%, ID 47%, II 22%). CONCLUSION: There is no relationship between progression towards ESRD and the ACE I/D polymorphism in ADPKD patients. PMID- 10831638 TI - Clinical manifestations of AB-amyloidosis: effects of biocompatibility and flux. AB - BACKGROUND: Highly permeable biocompatible dialysis membranes may postpone the development of AB-amyloidosis, but the relative contribution of enhanced flux or reduced inflammation by highly biocompatible membranes and sterile dialysis fluid remains unknown. METHODS: In this retrospective investigation, 89 patients with end-stage renal disease maintained on regular haemodialysis for at least 10 years and treated with one type of dialysis membrane exclusively were selected for analysis. They were divided into three groups: low-flux, bioincompatible cellulose (I), low-flux, intermediately biocompatible polysulphone or PMMA (II), or high-flux, highly biocompatible polysulphone or AN69 (III). In addition, the patients were analysed according to the microbiological quality of the dialysis fluid, which had been tested regularly and was classified either as standard or as intermittently contaminated. The clinical manifestations indicative of AB amyloidosis, namely, carpal tunnel syndrome, arthropathy and bone cysts, were diagnosed after recruitment. RESULTS: Clinical symptoms were most pronounced in group I, intermediate in group II, and lowest in group III. Patients treated with intermittently contaminated dialysis fluid showed a higher prevalence of AB amyloidosis than patients with less contaminated dialysis fluid. Logistic regression analysis demonstrated that the flux characteristics of the dialyser and the microbiological quality of the dialysis fluid as well as the biocompatibility of the dialyser were independent determinants of AB-amyloidosis. CONCLUSION: It would be prudent clinical practice to employ high-flux biocompatible membranes in conjunction with ultrapure dialysis fluid for the treatment of end-stage renal disease patients who need to remain on long-term haemodialysis. PMID- 10831639 TI - Free serum leptin but not bound leptin concentrations are elevated in patients with end-stage renal disease. AB - BACKGROUND: Leptin is a 16-kDa protein that is thought to be a regulator of food intake and body weight. Although total serum leptin levels have been reported to be elevated in obese and normal weight patients with end-stage renal disease (ESRD), it is not known whether serum-free leptin concentrations are also increased in patients with ESRD with no apparent nutritional problems. Furthermore, there are no data on how different dialysis modes (high-flux haemodiafiltration and low-flux dialysis) influence serum leptin subfractions. METHODS: We measured fasting serum free and bound leptin levels in three groups of male subjects: patients on haemodiafiltration with high flux dialysers (n=11), patients on haemodialysis with low-flux dialysers (n=17) and healthy age (61+/-8 years) and BMI (23.8+/-3.1 kg/m(2)) matched control subjects (n=28). Both leptin components were determined before and after a single dialysis session. RESULTS: Body mass indices were correlated with serum free leptin levels in both patients (r=0.69, P<0.001) and controls (r=0.77, P<0.001). Mean (SD) serum free leptin levels were significantly higher in ESRD patients than in control subjects (91+/ 33 vs 41+/- 21 pmol/l; P<0.01). Bound leptin levels did not differ in both groups (0.67+/-0.12 vs 0.56+/-0.11 nmol/l, NS). Elevated serum-free leptin levels in ESRD patients could be reduced by haemodiafiltration with high-flux membranes, but not with low-flux haemodialysis membranes. The former led to a reduction of initial serum free leptin values to 76+/-17% (P<0.01), whereas bound leptin remained unaffected. CONCLUSION: Serum-free leptin levels are elevated in ESRD without any apparent effect on body weight. In contrast, serum bound leptin levels remain stable, thus central feedback regulation via the bound form of the hormone may serve as an alternative explanation in the regulation of food intake and energy expenditure in chronic patients on haemodialysis with no apparent nutritional problems. PMID- 10831640 TI - The significance of serum homocysteine levels in diabetic patients on haemodialysis. AB - BACKGROUND: Atherosclerotic diseases are the major cause of mortality and morbidity in patients on haemodialysis (HD). Furthermore, the prognosis of diabetic patients on HD is especially poor due to atherosclerotic complications. Because homocysteine (Hcy), a sulfur-containing amino acid, is emerging as an important risk factor for atherosclerosis in patients with end-stage renal disease, we examined the significance of serum Hcy levels in diabetic patients on HD. METHODS: We measured total serum Hcy levels (tHcy) in 31 patients with diabetes mellitus on HD (DM group) and 37 non-diabetic patients on HD (N group), adjusting for age and HD duration. Linear regression analysis was used to assess the correlation of multiple variables to tHcy. RESULTS: The proportion of atherosclerotic disease in the DM group was significantly higher than in the N group. However, serum tHcy, serum creatinine and per cent creatinine generation rate in the DM group were significantly lower than in the N group. In the DM group, serum tHcy was positively correlated with creatinine, albumin and per cent creatinine generation rate, respectively. This was not the case in the N group. CONCLUSIONS: The demethylation pathway in methionine metabolism in the liver, which is linked directly to the creatinine generation system, may be disturbed in diabetic patients on HD. This may be the reason why serum tHcy and creatinine in diabetic patients on HD are lower than in non-diabetic patients on HD. Therefore, it is necessary to consider the possibility of an altered relation between serum tHcy and vessel disease when evaluating the atherogenic risk in diabetic patients on HD. PMID- 10831641 TI - Short-term recombinant human growth hormone therapy does not modify growth hormone, thyrotropin and prolactin responses to thyrotropin-releasing hormone in adult dialysis patients. AB - BACKGROUND: We recently have reported the first randomized, controlled study on the effects of short-term recombinant human growth hormone (rhGH|| therapy on the nutritional status of a group of malnourished adult dialysis patients. In order to evaluate whether rhGH administration exerts any influence on GH, thyrotropin (TSH|| and prolactin (PRL|| responses to TSH-releasing hormone (TRH||, we assessed these responses before and after rhGH therapy. METHODS: GH, PRL and TSH responses to TRH before and 1 month after rhGH therapy in a group of adult dialysis patients were evaluated. Seventeen dialysis patients (11 on continuous ambulatory peritoneal dialysis/six on haemodialysis|| were studied (rhGH group, n=8; control group, n=9||. In the rhGH group, 0.2 IU/kg/day rhGH was administered subcutaneously. Each patient was tested with TRH (400 microg bolus i.v.|| on two separate occasions, just before and immediately after the treatment period. RESULTS: rhGH treatment did not modify baseline serum GH concentrations (6.6+/ 2.7 vs 4.1+/-1.1 microg/l||, paradoxical GH responses to TRH (six out of eight patients||, GH peak (11.9+/-4.6 vs 11.2+/-5.3 microg/l, NS|| or area under the secretory curve of GH (GH AUC; 19.1+/-4.5 vs 12.1+/-3.1 microg/h/l||. Both basal PRL (35.5+/-7.1 vs 36.7+/-8.6 microg/l|| and TSH (2.3+/-1.1 vs 2.8+/-1.7 mU/l|| concentrations, as well as their responses to TRH stimulation (PRL peak, 59.9+/ 16.6 vs 59. 5+/-11.8 microg/l; TSH peak, 6.2+/-2.6 vs 7.1+/-3.9 mU/l||, were also unaffected by rhGH therapy. CONCLUSION: These results suggest that short-term rhGH therapy does not significantly influence the magnitude of the somatotropic, lactotropic or thyrotropic response to TRH in adult dialysis patients. However, this finding has to be interpreted with caution due to the two different patient groups included in this study. PMID- 10831642 TI - Effects of icodextrin in automated peritoneal dialysis on blood pressure and bioelectrical impedance analysis. AB - BACKGROUND: Glucose absorption from glucose-based dialysis fluids limits ultrafiltration from the daytime dwell in automated peritoneal dialysis (APD). Icodextrin may allow greater ultrafiltration during the daytime period in APD, enhancing fluid control. METHODS: A 7.5% icodextrin dialysate was compared with a 2. 27% glucose dialysate for the daytime dwell in 14 subjects on APD. Blood pressure, weight and body water compartments estimated by multifrequency bioelectrical impedance (MFBIA) were determined in subjects using 2.27% glucose as the daytime dwell and then repeated 1 month after switching to icodextrin. RESULTS: Icodextrin resulted in symptomatic hypotension requiring reduction of antihypertensive medication in six of the 14 patients. Despite this reduction in treatment, systolic blood pressure fell from 142.4 (23.9) mmHg to 122.9 (17.7) mmHg, P<0.005, and diastolic blood pressure tended to fall from 82.8 (9.8) mmHg to 76.8 (10.1) mmHg, P=0.075. Change in systolic blood pressure significantly correlated with changes in weight (r=0.62, P<0.05) and MFBIA estimates of total body water (TBW) (r=0.56, P<0.05), extracellular water (ECW) (r=0.79, P<0.002), extra/intracellular water ratio (ECW/ICW) (r=0.72, P<0.01) and derived resistances R(ecf) of ECW (r=-0.69, P<0.01) and R(inf) of TBW (r=-0.66, P<0.02). Changes in diastolic blood pressure significantly correlated with changes in ECW (r=0.64, P<0.02) and ECW/ICW ratio (r=0.58, P<0.05), and almost significantly with R(ecf) (r=-0.51, P=0.08) and R(inf) (r=-0.52, P=0.07) estimated by MFBIA, but not with changes in weight or TBW. CONCLUSIONS: Use of icodextrin for the daytime dwell in APD results in improved fluid balance and blood pressure control compared with 2.27% glucose. MFBIA detected clinically important changes in fluid content in these patients. PMID- 10831643 TI - Tacrolimus reversibly reduces insulin secretion in paediatric renal transplant recipients. AB - BACKGROUND: Conflicting reports exist about the mechanism of tacrolimus-induced post-transplant diabetes mellitus. METHODS: We analysed intravenous glucose tolerance tests (IVGTT) of 14 paediatric renal transplant recipients on cyclosporin (CsA) microemulsion and 15 patients on tacrolimus (FK506). The groups were similar in age (13.2+/-4.2 vs 13.0+/-3.7 years), body mass index, serum creatinine concentrations (96+/-60 vs 97+/-44 micromol/l), time after renal transplantation, and cumulative steroid dose over 12 weeks prior to the test (3.4 vs 3.5 mg/m(2)/day, NS, Mann-Whitney). Parameters of glucose tolerance included glucose, insulin, C-peptide concentrations, and HbA1c. The mean concentrations of the primary immunosuppressant were similar to treatments employed in other centres (CsA 165+/-59 ng ml and FK506 7. 5+/-2.2 ng ml). RESULTS: Baseline glucose concentrations were significantly higher on FK506 therapy compared with CsA microemulsion therapy. Baseline insulin concentrations and C-peptide concentrations were identical in both treatment groups. FK506 trough levels correlated negatively with k values (glucose constant decay) in the FK506 group. There was a significant reduction of the insulin first-phase concentrations, both after 1 min and after 3 min in the FK506 group compared with the CsA group (112+/ 17 vs 237+/-57 microU/ml, P=0.034). In patients with repetitive IVGTTs, glucose constant decay and insulin production improved after lowering FK506 whole-blood trough levels. CONCLUSIONS: We conclude that post-transplant glucose intolerance could be due to a dose-dependent, direct effect of FK506 on the pancreatic beta cell function, which can be controlled by dose reduction. PMID- 10831644 TI - Platelet-derived growth factor, basic fibroblast growth factor, and interferon gamma increase type IV collagen production in human fetal mesangial cells via a transforming growth factor-beta-dependent mechanism. AB - BACKGROUND: Glomerulosclerosis is characterized by glomerular accumulation of extracellular matrix following mesangial cell proliferation. The precise pathomechanism of glomerulosclerosis is still undetermined. Platelet-derived growth factor (PDGF) and basic fibroblast growth factor (b-FGF) are known to be mitogenic for mesangial cells, and interferon gamma (IFN-gamma) is known to have an inhibitory effect on mesangial cell proliferation. We attempted to clarify the role of these cytokines on mesangial matrix production using cultured human fetal mesangial cells (HMC). METHODS: HMC were incubated with these cytokines for 24-72 h and the levels of type IV collagen and TGF-beta in the cell supernatants were measured by enzyme immunoassay. RESULTS: PDGF, b-FGF, and IFN-gamma stimulated type IV collagen production by HMC in a dose- and time-dependent manner. The anti TGF-beta neutralizing antibody clearly inhibited their stimulatory effect on type IV collagen production. PDGF and b-FGF also stimulated TGF-beta production by HMC in a dose-dependent manner, although IFN-gamma did not. CONCLUSION: PDGF, b-FGF, and IFN-gamma stimulate type IV collagen production in cultured HMC via a TGF beta-dependent mechanism. PMID- 10831645 TI - Changes in bone turnover after parathyroidectomy in dialysis patients: role of calcitriol administration. AB - BACKGROUND: Available data on changes in serum levels of bone markers after parathyroidectomy (PTx) in dialysis patients are not uniform. Changes are thought to be due to either a reduction in PTH activity per se or to a direct effect of vitamin D therapy on bone cells. We aimed to verify whether treatment with vitamin D modifies serum levels of markers of bone synthesis (alkaline phosphatase (AP), osteocalcin (BGP), procollagen type I C-terminal peptide (PICP)) and resorption (collagen type I C-terminal peptide (ICTP)) within a period of 15 days in haemodialysis patients with severe secondary hyperparathyroidism following PTx. METHODS: We randomized two groups (A, treatment and B, placebo, 10 patients each) with comparable basal PTH values and measured bone markers 3, 7 and 15 days after surgery. All patients were treated with calcium supplements (i.v. and p.o.), and group A also received calcitriol (2.4+/-1.0 microg/day, p.o.). RESULTS: In both groups, PTx induced significant changes in all the markers evaluated, except for BGP in group B. Compared to basal values, ICTP decreased from 481+/-152 ng/ml in group A and 277+/-126 ng/ml in group B to 267+/-94 and 185+/-71 ng/ml (M+/-SD) respectively, and PICP increased from 307+/-139 ng/ml in group A and 309+/-200 ng/ml in group B to 1129+/-725 and 1231+/-1267 ng/ml (M+/-SD) respectively, within 3 days of surgery. AP values increased after 15 days from 1115+/-734 mU/ml in group A and 1419+/ 1225 mU/ml in group B to 1917+/-1225 and 1867+/-1295 mU/ml (M+/-SD) respectively. On the contrary, mean values of BGP were never different from basal levels after PTx in either group. In the two groups, the pattern of changes of all the bone markers after PTx was almost identical. Group A patients predictably required lower doses of oral calcium supplements to correct hypocalcaemia (16. 9+/-5.7 vs 22.1+/-5.0 g/10 days; M+/-SD, P<0.04). CONCLUSIONS: The opposite behaviour of serum PICP and ICTP after PTx, in both the treated and untreated groups suggests that quantitative uncoupling between bone synthesis and resorption is responsible for hypocalcaemia. This phenomenon, as reflected by the evaluated bone markers, is unaffected by calcitriol. Based on our data we conclude that immediately after parathyroid surgery, vitamin D therapy does not influence bone cell activity, but improves hypocalcaemia mainly through its known effect on intestinal calcium absorption. PMID- 10831646 TI - Haemolysis in haemodialysis patients: evidence for impaired defence mechanisms against oxidative stress. AB - BACKGROUND: Uraemic patients have a decreased ability to withstand oxidative stress. It is postulated that their antioxidant capacity is reduced, yet the mechanism remains unclear. Recently 33 haemodialysis (HD) patients were exposed to chloramine contamination in the water supply. This led to haemolysis in 24 patients, while nine were unaffected. In the former group haemoglobin decreased from 11.7+/-1.1 to 8.5+/- 1.4 g/dl (P<0.0001) and returned to 11.4+/-0.9 g/dl (P<0.0001) following recovery. During haemolysis, haptoglobin was 38.4+/-10.6 vs 138.1+/-8.3 ng/dl (P<0.0001) following recovery. METHODS: To explore the factors affecting the severity of haemolysis we studied extracellular and intracellular anti-oxidant defence mechanisms 3 months after recovery. In 29 patients and 20 controls we determined plasma glutathione (GSH), and the erythrocyte enzymes glutathione peroxidase (GSH-Px), glutathione reductase (GSH-Rx), and superoxide dismutase (SOD). Serum malondialdehyde (MDA) was measured as a marker of oxidative stress. RESULTS: Plasma GSH was lower in patients as compared to controls (5.49+/-0.26 vs 7.4+/-0.5 micromol/l, P<0.005). There was an inverse correlation between GSH and the degree of haemolysis (r=-0.42, P<0.02). Patients had higher GSH-Rx (4.64+/-0.15 vs 3.97+/-0.12 U/gHb, P<0.02), lower GSH-Px (29. 7+/-1.85 vs 35.5+/-1.62 U/gHb, P<0.001), and similar SOD (0.63+/-0. 02 vs 0.51+/ 0.02 U/mgHb) as compared to controls. There was no correlation between the enzyme levels and the degree of haemolysis. MDA was higher in patients (2.37+/-0.07 vs 0.97+/-0.1 nmol/ml, P<0. 0001). There was a correlation between MDA and the years patients were on HD (r=0.43, P<0.02). CONCLUSIONS: These data indicate that HD patients have an impaired anti-oxidant response, which may be attributed in part, to plasma GSH deficiency. Patients with the lowest plasma GSH levels are more susceptible to oxidative stress and consequent haemolysis. PMID- 10831647 TI - Immunocytological determination of lymphocytes and monocytes/macrophages in urinary sediments of renal allograft recipients. AB - BACKGROUND: Urinary studies using Papanicolaou staining following kidney transplantation led to the conjecture that acute allograft rejection might be accompanied by an increased lymphocyturia. However, it is difficult to distinguish lymphoid cells from other urinary cells using conventional stains. METHODS: Staining of urinary lymphocytes using FITC-labelled antibodies is complicated by a high unspecific fluorescence that limits the evaluation. Therefore, we developed a method to stain urinary lymphocytes using enzyme-linked antibodies. The cells were cytocentrifuged onto microscope slides and were fixed. RESULTS: By means of a combined evaluation of Papanicolaou and immunocytochemical staining, CD3-positive pan T cells, CD4-positive T-helper cells, CD8-positive cytotoxic/suppressor cells, and CD14-positive monocytes/macrophages of urinary sediments were determined in 41 kidney graft recipients following renal transplantation. During periods of normal graft function, neither positive lymphocytes nor positive monocytes/macrophages were found in the urinary sediments. However, in the course of acute allograft rejection a significant increase in positive lymphocytes and positive monocytes/macrophages could be observed. Interestingly, in cases of acute allograft rejection the distribution of urinary lymphocytes and monocytes was comparable to the distribution of infiltrating immunocompetent cells in renal allograft biopsies. CONCLUSION: The present study demonstrates that immunocytochemical staining via enzyme-conjugated antibodies is a reliable method to visualize T lymphocytes and monocytes/macrophages in the urinary sediment, and that this technique may be of special diagnostic value in the diagnosis of acute allograft rejection. PMID- 10831648 TI - Vasculitic wallenberg syndrome with detection of anti-proteinase 3 antibodies in the cerebrospinal fluid of a patient with severe Wegener's granulomatosis and only mild kidney involvement. PMID- 10831649 TI - Successful treatment of recurrence of immunotactoid glomerulopathy in a kidney allograft recipient. PMID- 10831650 TI - Catheter fracture-an underrecognized and serious condition in haemodialysis. PMID- 10831651 TI - Gross left-sided hydronephrosis in a renal transplant recipient. PMID- 10831652 TI - The patient with Wegener's granulomatosis and an intrasplenic mass of unknown origin. PMID- 10831653 TI - Cutaneous white spots in a child with polycystic kidneys: a clue to TSC2/PKD1 gene mutation. PMID- 10831654 TI - The renal sinus cyst-the great imitator. PMID- 10831656 TI - Berndt tersteegen award 2000 PMID- 10831655 TI - Update in nephrology Beograd, 12 February 2000. PMID- 10831657 TI - A patient with heat insoluble cryoglobulin. PMID- 10831658 TI - Anti-oxidant and mushroom poisoning. PMID- 10831659 TI - A patient with heat insoluble cryoglobulin PMID- 10831660 TI - (CA)(n) dinucleotide repeat polymorphism at the 5'-end of the aldose reductase gene is not associated with microangiopathy in Caucasians with long-term diabetes mellitus 1. PMID- 10831661 TI - Goodpasture's syndrome treated with mycophenolate mofetil. PMID- 10831662 TI - The patient with over 100 relapses of minimal change nephrotic syndrome: prolonged complete remission after chlorambucil treatment. PMID- 10831663 TI - Massive proteinuria in a patient with chronic pyelonephritis. PMID- 10831664 TI - The patient with over 100 relapses of minimal change nephrotic syndrome: prolonged complete remission after chlorambucil treatment PMID- 10831665 TI - Nephrotic syndrome associated with isotretinoin. PMID- 10831666 TI - Atheromatous renal artery stenosis rarely causes renal failure. PMID- 10831667 TI - Biocompatible dialysis membranes do not reduce plasma leptin levels. PMID- 10831668 TI - Timing of sildenafil therapy in dialysis patients-lessons following an episode of hypotension. PMID- 10831669 TI - A new form of acute adverse reaction to icodextrin in a peritoneal dialysis patient. PMID- 10831670 TI - Post-transplantation lymphoproliferative disorder: an unusual presentation in a patient receiving tacrolimus. PMID- 10831671 TI - Hyperlipidaemia and post-heparin lipase activities in renal transplant recipients treated with sirolimus or cyclosporin A. PMID- 10831672 TI - Post-transplantation lymphoproliferative disorder: an unusual presentation in a patient receiving tacrolimus PMID- 10831673 TI - Announcements PMID- 10831674 TI - Radiology 2000: reviewing for radiology PMID- 10831675 TI - Obstetric US imaging: the past 40 years. AB - Although diagnostic ultrasonography (US) was developing in the late 1940s and early 1950s, it was not until the 1960s, with the availability of commercial equipment, that its usefulness in obstetrics began to be realized fully by radiologists and obstetricians around the world. Advances from A-mode to bistable and then to gray-scale static imaging were followed by the introduction of automated compound imaging and real-time US. Also, the development and initial use of Doppler US for the detection of fetal heart motion and the eventual use of pulsed and color Doppler US for the evaluation of such fetal structures as the major vessels and heart chambers contributed to increasing the usefulness of US in obstetrics. The development of specialized transducers--in particular, endovaginal probes--resulted in images of the early fetus. At the present time, the development of multiplanar, three-dimensional imaging shows great promise for more complete imaging of the fetus. The importance of US in the examination of the pregnant patient and, in particular, of the fetus has led to its worldwide dominance as the imaging modality of choice. The contributions of obstetric US to improving maternal well-being and fetal health have been recognized as a key component in all countries around the world. PMID- 10831676 TI - The role of the supertechnologist. PMID- 10831677 TI - Biomedical imaging symposium: visualizing the future of biology and medicine. PMID- 10831678 TI - MR enteroclysis: the future of small-bowel imaging? PMID- 10831679 TI - Double-contrast barium enema examination technique. AB - This review article presents the principles for performing a safe, comfortable, and accurate double-contrast barium enema examination. The procedure is a flexible examination in which the fluoroscopist interacts with the patient, the controls of the fluoroscope, and the image on the television monitor. During a double-contrast examination, images of the colon are created by manipulating the patient, the barium pool, and the amount of air insufflated into the rectum. Fluoroscopy is essential for guiding the radiologist to obtain spot images with adequate technical factors. The fluoroscopist analyzes the luminal contour, the barium-coated mucosal surface en face, and the barium pool to detect abnormalities in the colon. With careful technique, a high-quality examination can be performed in most patients. PMID- 10831680 TI - Hemodialysis catheter placement and management. AB - Hemodialysis catheters are an integral part of the delivery of hemodialysis. While catheters play an important role in the patient undergoing hemodialysis, catheters should be considered a bridge to more permanent forms of dialysis access in most patients. Recent advances in catheter technology, access techniques, and choice of access sites have improved outcomes associated with hemodialysis catheters. The placement and management of hemodialysis catheters by interventional radiologists have played an important role in these advances, and interventional radiologists are taking an increasingly active role in the research and development of catheters and catheter insertion techniques. The present status of hemodialysis catheters is reviewed. PMID- 10831681 TI - Advances in gastrointestinal intervention: the treatment of gastroduodenal and colorectal obstructions with metallic stents. AB - Metallic stents are currently an established component of the endoluminal treatment of stenoses within the blood vessels, bile ducts, esophagus, trachea, and bronchi. With the development of newer stent designs and delivery systems and the general momentum toward minimally invasive therapies, metallic stent placement has expanded into the nonsurgical therapy for gastroduodenal and colorectal obstructions. The use of metallic stents within the stomach, duodenum, or colon is intended not to be curative but to provide nonsurgical palliation for the symptoms of gastric or colonic obstruction. This palliation may be intended for the life of the patient in the case of unresectable disease or as a temporizing procedure prior to a definitive surgical procedure. In the latter clinical scenario, the benefits of a minimally invasive intestinal decompression procedure include (a) quick and noninvasive relief of the intestinal obstruction in an acutely ill patient that obviates a more extensive procedure; (b) allowance of time to improve a patient's overall medical condition and thus to allow a patient to better tolerate the definitive surgical procedure; and (c) reduction of the complexity of the definitive procedure by eliminating the need for staged procedures and allowing the definitive procedure to be performed at one setting. PMID- 10831682 TI - Aorta and iliac arteries: single versus multiple detector-row helical CT angiography. AB - PURPOSE: To compare single- versus four-channel helical computed tomographic (CT) aortography. MATERIALS AND METHODS: Forty-eight patients with aortic aneurysm or dissection underwent four- and one-channel CT angiography. Scan pairs covered the thoracic inlet to the diaphragm (n = 10) and supraceliac abdominal aorta (n = 19) or thoracic inlet (n = 19) to the femoral arterial bifurcations. For four-channel CT, nominal section thickness and pitch were 2.5 mm and 6.0, respectively, and for one-channel CT, 3.0 mm and 2.0 to the infrarenal aorta and 5.0 mm and 2.0 to the femoral arteries. Effective section thickness, scanning duration, scanning coverage, dose of iodinated contrast material, and mean aortoiliac attenuation were compared. Data were summarized as speed (coverage/duration), scanning efficiency (speed/section thickness), and contrast efficiency (mean aortic attenuation/dose of contrast material). RESULTS: At four- versus one-channel CT, CT angiography was 2.6 times faster, scanning efficiency was 4.1 times greater, contrast efficiency was 2.5 times greater, dose of contrast material was reduced (mean, 57%; 97 vs 232 mL) without a significant change in aortic enhancement, and sections were thinner (mean, 40%; 3.2 vs 5.3 mm) despite a 59% shorter scanning duration (22 vs 56 seconds). CONCLUSION: Substantially reduced doses of contrast medium, shorter scanning durations, and narrower effective sections result with four- versus one-channel CT aortography. No advantages of one-channel CT aortography were demonstrated. PMID- 10831683 TI - Postprocedural hypotension after carotid artery stent placement: predictors and short- and long-term clinical outcomes. AB - PURPOSE: To describe the predictors of persistent hypotension after carotid artery stent (CAS) placement and define the clinical outcome of patients with this hemodynamic disturbance. MATERIALS AND METHODS: One hundred forty CAS procedures were performed in 133 consecutive patients. Post-CAS hypotension defined as a greater than 40 mm Hg decrease in arterial pressure without evidence of hypovolemia, with a systolic pressure lower than 90 mm Hg at the end of CAS and lasting at least 1 hour-was observed in 25 patients (group 1); 108 patients did not have hypotension (group 2). RESULTS: Post-CAS hypotension developed in 33.9% of cases after balloon-expandable stent placement versus in 13.6% of cases after self-expanding stent placement (P =.04). In-hospital minor ipsilateral strokes occurred in 16% of cases in group 1 versus in 3% of cases in group 2 (P =.03). There was one (0.9%) major stroke (transient) and three (2.6%) transient ischemic attacks, all of which occurred in group 2 (not significant vs group 1 for both conditions). At 10 months +/- 4 (SD) of follow-up, there was greater total mortality in group 1 than in group 2 (20% vs 4%, P =.02), whereas neurologic events did not differ significantly between the groups. CONCLUSION: Hypotension due to carotid sinus stimulation is frequent after CAS with balloon expandable stents. This phenomenon correlates with increased in-hospital complications and long-term risk of death. PMID- 10831684 TI - Interventional radiologic procedures: patient anxiety, perception of pain, understanding of procedure, and satisfaction with medication--a prospective study. AB - PURPOSE: To prospectively assess patient anxiety, understanding of the procedure being performed, perception of pain level, and satisfaction with medication given for a variety of diagnostic and therapeutic vascular and visceral (nonvascular) interventional procedures. MATERIALS AND METHODS: The authors interviewed 204 patients before and after they underwent an interventional radiologic procedure. Patients responded to a series of questions by using a visual analog scale. Patients were grouped according to (a) their level of experience with the procedure and (b) the type of procedure performed (diagnostic or therapeutic visceral procedure or diagnostic or therapeutic vascular procedure). RESULTS: Patients who had previous experience with a procedure, whether visceral or vascular, were less anxious, had more understanding, and anticipated less pain than did those who did not have experience with a procedure. Patients who had only local anesthesia for visceral biopsy experienced greater pain than did those who had both local and intravenous anesthesia. Satisfaction scores, however, were similar throughout all groups. CONCLUSION: Patients have a moderate amount of anxiety about interventional procedures and anticipate some discomfort. Most patients have a high level of satisfaction despite the amount of pain they experience during the procedure. Patients experienced with a procedure tend to have a greater understanding of the procedure and less anxiety. PMID- 10831685 TI - Transjugular versus percutaneous renal biopsy for the diagnosis of parenchymal disease: comparison of sampling effectiveness and complications. AB - PURPOSE: To compare the effectiveness and safety of transjugular renal biopsy with those of percutaneous renal biopsy for diagnosis of renal parenchymal disease. MATERIALS AND METHODS: Results and complications of 400 consecutive transjugular renal biopsies performed between 1993 and 1998 with a modified Colapinto transjugular hepatic biopsy system were compared retrospectively with those of 400 percutaneous renal biopsies performed during the same period. Transjugular renal biopsy was associated with 14 cardiac and 35 hepatic biopsies. Number of glomeruli per tissue core, adequacy of tissue core for histopathologic diagnosis, and rate and severity of complications were analyzed. RESULTS: Renal tissue was obtained with percutaneous renal biopsy in 382 (95.5%) of 400 patients and with transjugular renal biopsy in 383 (95.8%) of 400 patients. The mean numbers of intact glomeruli per tissue core with optical microscopy were 11.2 +/- 7.7 (SD) and 9.8 +/- 7.6 for percutaneous renal biopsy and transjugular renal biopsy, respectively. With immunofluorescent microscopy, the mean numbers were 6.4 +/- 5.3 and 4.6 +/- 4.6 for percutaneous renal biopsy and transjugular renal biopsy, respectively. Tissue cores were adequate for histopathologic diagnosis in 98.2% with both techniques. Major complications occurred with transjugular renal biopsy in four patients and with percutaneous renal biopsy in three patients. CONCLUSION: Use of transjugular renal biopsy provides diagnostic yield and safety similar to those of percutaneous renal biopsy and allows multiorgan biopsy during the same procedure. It can be recommended in patients with percutaneous renal biopsy contraindication or failure. PMID- 10831686 TI - Accuracy and complication rates of US-guided vacuum-assisted core breast biopsy: initial results. AB - PURPOSE: To determine the accuracy and complications of vacuum-assisted core breast biopsy performed with ultrasonographic (US) guidance. MATERIALS AND METHODS: US-guided, vacuum-assisted breast biopsy with an 11-gauge device was performed in 71 lesions in 67 consecutive women (age range, 23-82 years; mean age, 52.9 years). Vacuum-assisted core biopsy findings were compared with excisional biopsy, mammographic follow-up, and clinical follow-up findings (follow-up, 1-19 months; mean, 9.2 months). Procedural complications and treatment were noted. RESULTS: Of 71 lesions, 18 (25%) were diagnosed as malignant at core biopsy; one (1%), as premalignant; 30 (42%), as specific benign; and 22 (31%), as nonspecific benign. Of 18 malignant diagnoses, one (6%) was benign at excision. The premalignant specimen was benign at excision. Of 52 benign findings, 51 (98%) were proved benign at excision, mammographic follow-up, or clinical follow-up. One benign finding was carcinoma at excision. In this case, the specimen did not include the carcinoma because of a technical problem recognized at the time of the percutaneous procedure. Five (7%) of 71 biopsies resulted in bleeding beyond 10 minutes. One (1%) patient experienced a vasovagal response. CONCLUSION: According to these data, US-guided vacuum-assisted core breast biopsy is accurate. There may be a slightly higher risk of bleeding, which may be related to the lack of breast compression during the procedure, when compared with biopsy performed with stereotactic guidance. PMID- 10831687 TI - Reasons for failure of a mammography unit at clinical image review in the American College of Radiology Mammography Accreditation Program. AB - PURPOSE: To identify the most common deficiencies in the quality of mammograms submitted for clinical image evaluation (evaluation of image from actual patient referred for mammography). MATERIALS AND METHODS: In 1997, the American College of Radiology Mammography Accreditation Program reviewed clinical images for 2,341 mammography units. For each mammography unit, the facility submitted bilateral mediolateral oblique and craniocaudal mammograms obtained in a woman with fatty breasts and a woman with dense breasts. Images were reviewed independently by two experienced radiologists. Reviewers listed the general categories and specific deficiencies that led to a decision to fail the unit that produced the clinical images. RESULTS: Of the 2,341 mammography units, 1,034 (44%) failed the clinical image evaluation process. Of 6,128 categories cited by reviewers as deficient, 1,250 (20%) involved problems in positioning; 944 (15%), exposure; 887 (14%), compression; 806 (13%), sharpness; 785 (13%), contrast; 703 (11%), labeling; 465 (8%), artifacts; and 288 (5%), noise. A significantly higher proportion of failures was attributed to positioning deficiencies for fatty breasts than for dense breasts (P =.028). Higher proportions of failures in dense breasts were related to compression (P <.001) and exposure (P <.001) deficiencies. CONCLUSION: Common problems in clinical image quality have been identified. This information should be useful for educators and facilities striving to improve the quality of mammography. PMID- 10831688 TI - Breast cancer: importance of spiculation in computer-aided detection. AB - PURPOSE: To determine the prevalence of spiculation in a large series of screening-detected breast cancers appearing as masses on mammograms and to assess the sensitivity of a computer-aided detection (CAD) algorithm that uses spiculation measures in the detection of such lesions. MATERIALS AND METHODS: Six hundred seventy-seven consecutive cases of breast cancers detected as masses on mammograms were independently reviewed by three radiologists who determined if the lesions were spiculated. All cancers were then analyzed by the CAD system. RESULTS: All three radiologists interpreted 375 (55%) of the 677 masses as being spiculated on at least one view. The CAD algorithm correctly marked 322 (86%) of the 375 clearly spiculated masses, with a mean of 0.24 additional mass mark per image. With a looser definition of spiculation, 585 (86%) of the 677 masses were called spiculated by at least one radiologist on one view. The algorithm correctly marked 464 (79%) of the 585 lesions that were spiculated or possibly spiculated. CONCLUSION: Spiculation was clearly present in a majority (55%) of consecutive screening-detected breast cancer masses found on mammograms in a large clinical trial. Incorporation of spiculation measures is, therefore, an important strategy in the detection of breast cancer with CAD. A present generation CAD algorithm correctly identified a large proportion (86%) of spiculated breast cancers. PMID- 10831689 TI - Activity-based cost analysis: a method of analyzing the financial and operating performance of academic radiology departments. AB - PURPOSE: To develop a methodology for an activity-based cost (ABC) analysis in an academic radiology department, to test the hypothesis that the business of academic radiology can be separated into three distinct businesses-clinical activity, teaching, and research-and to determine the effect of the current teaching paradigm on clinical productivity. MATERIALS AND METHODS: Forty-seven key departmental activities were defined and distributed among the teaching, research, and clinical businesses. Individual radiologists determined the time spent in each of these activities by completing a detailed log of every activity performed during 2 weeks. All departmental revenue and costs were assigned to each activity in each of the three businesses. RESULTS: The methodology provided a successful understanding of the relative costs of each of the businesses of teaching, research, and clinical activity. It also provided the departmental costs of performing the separate activities typical of each business. Key findings included the following: Faculty spends 72% of time in clinical activities, research is the most expensive service per direct activity hour, and clinical reads (23%) are the single largest departmental cost element. CONCLUSION: ABC analysis can separate academic radiology into three businesses teaching, research, and clinical-and provide a detailed understanding of the cost structure of each. This analysis identifies opportunities for improved quality of service, productivity, and cost within each business. PMID- 10831690 TI - Small-bowel disease: comparison of MR enteroclysis images with conventional enteroclysis and surgical findings. AB - PURPOSE: To investigate if magnetic resonance (MR) enteroclysis can be performed routinely and to compare MR enteroclysis findings with those of conventional enteroclysis or surgery. MATERIALS AND METHODS: MR enteroclysis was prospectively performed in 30 patients with symptoms of inflammatory bowel disease or small bowel obstruction (SBO). A methylcellulose-water solution was used to distend the small bowel. To monitor dynamic changes in the small bowel, a single-shot fast spin-echo T2-weighted sequence was applied. For morphologic assessment, breath hold T2-weighted fast spin-echo and coronal T1-weighted gradient-recalled-echo MR images were obtained without and with gadolinium enhancement. Image quality and degree of small-bowel distention were graded. MR imaging findings and degree of SBO were compared with findings at conventional enteroclysis (n = 25) or surgery (n = 5). RESULTS: MR enteroclysis was well tolerated and provided adequate image quality and sufficient small-bowel distention. SBO grade based on MR enteroclysis images (n = 10) was identical to that based on conventional enteroclysis images (n = 6) or surgical findings (n = 4). There was exact agreement between MR enteroclysis and retrospective findings in all five patients who underwent surgery, and MR findings were identical to those at enteroclysis in 18 patients, superior in six patients, and inferior in one patient. CONCLUSION: MR enteroclysis can be performed routinely with adequate image quality and sufficient small-bowel distention. The functional information provided by MR enteroclysis is identical to that provided at conventional enteroclysis. PMID- 10831691 TI - Focal liver lesions: evaluation of the efficacy of gadobenate dimeglumine in MR imaging--a multicenter phase III clinical study. AB - PURPOSE: To evaluate gadobenate dimeglumine (Gd-BOPTA) for dynamic and delayed magnetic resonance (MR) imaging of focal liver lesions. MATERIALS AND METHODS: In 126 of 214 patients, MR imaging was performed before Gd-BOPTA administration, immediately after bolus administration of a 0.05- mmol/kg dose of Gd-BOPTA, and 60-120 minutes after an additional intravenously infused 0.05-mmol/kg dose. In 88 patients, imaging was performed before and 60-120 minutes after a single, intravenously infused 0.1-mmol/kg dose. T1- and T2-weighted spin-echo and T1 weighted gradient-echo images were acquired. On-site and blinded off-site reviewers prospectively evaluated all images. Intraoperative ultrasonography, computed tomography (CT) during arterial portography, and/or CT with iodized oil served as the reference methods in 110 patients. RESULTS: Significantly more lesions were detected on combined pre- and postcontrast images compared with on precontrast images alone (P <. 01). All reviewers reported a decreased mean size of the smallest detected lesion and improved lesion conspicuity on postcontrast images. All on-site reviewers and two off-site reviewers reported increased overall diagnostic confidence (P <.01). Additional lesion characterization information was provided on up to 109 (59%) of 184 delayed images and for up to 50 (42%) of 118 patients in whom dynamic images were assessed. Gd-BOPTA would have helped change the diagnosis in 99 (47%) of 209 cases and affected patient treatment in 408 (23%) of 209 cases. CONCLUSION: Gd-BOPTA increases liver lesion conspicuity and detectability and aids in the characterization of lesions. PMID- 10831692 TI - Detection of choledocholithiasis with MR cholangiography: comparison of three dimensional fast spin-echo and single- and multisection half-Fourier rapid acquisition with relaxation enhancement sequences. AB - PURPOSE: To compare the performance of three pulse sequences commonly used at magnetic resonance (MR) cholangiography in the diagnosis of choledocholithiasis. MATERIALS AND METHODS: MR cholangiography was performed in 57 patients who were suspected of having choledocholithiasis and referred for endoscopic retrograde cholangiography. Non-breath-hold three-dimensional fast spin-echo, breath-hold single-section half-Fourier rapid acquisition with relaxation enhancement (RARE), and breath-hold multisection half-Fourier RARE sequences were compared. Two radiologists independently interpreted the MR cholangiograms. Evaluated diagnostic performance parameters included sensitivity, specificity, receiver operating characteristic (ROC) curves, and interobserver agreement (kappa statistics). Endoscopic retrograde cholangiography was the standard of reference. RESULTS: Eight patients were excluded because of incomplete MR examinations (n = 4) or failure in the cannulation of the bile duct at retrograde cholangiography (n = 4). In 49 patients, the three MR cholangiographic sequences were completed successfully. In 24 (49%) of these patients, retrograde cholangiography demonstrated stones. Sensitivity and specificity of MR cholangiography exceeded 90%, and the area under the ROC curve was greater than 0.95 for both radiologists and for the three sequences. Interobserver agreement for presence of bile duct stones was at least 0.80 (very good) for the three sequences. CONCLUSION: The three MR cholangiographic sequences had similarly high sensitivities and specificities for the detection of choledocholithiasis. PMID- 10831693 TI - Focal liver lesions: pattern-based classification scheme for enhancement at arterial phase CT. AB - PURPOSE: To present our early experience with a classification scheme for categorizing focal liver lesions on the basis of the enhancement patterns that they exhibit in the arterial phase of computed tomography (CT) and to determine whether particular enhancement patterns suggest particular diagnoses. MATERIALS AND METHODS: The authors reviewed arterial phase CT images in 100 consecutive patients with focal liver lesions, excluding simple cysts. The enhancement pattern of the dominant or representative lesion in each patient was classified into one of five categories-homogeneous, abnormal internal vessels or variegated, peripheral puddles, complete ring, or incomplete ring-by three radiologists blinded to the proved diagnosis. Lesions without enhancement were recorded separately. Agreement was reached by consensus in all cases. Standards of reference included findings at histologic examination, correlative imaging, or clinical and imaging follow-up. RESULTS: Ninety-two percent of the 100 lesions demonstrated arterial phase enhancement. Patterns associated with positive predictive values of 82% or greater and specificity of 80% or greater included abnormal internal vessels or variegated (hepatocellular carcinoma), peripheral puddles (hemangioma), and complete ring (metastasis). CONCLUSION: The appearance of hepatic lesions in the arterial phase of enhancement has potential use in the determination of specific diagnoses. The classification scheme used in this study may be a useful tool for the interpretation of arterial phase CT studies. PMID- 10831694 TI - Case 27 PMID- 10831695 TI - Case 23: replacement lipomatosis of the kidney. PMID- 10831696 TI - Prostate-specific antigen: effect of pelvic irradiation. AB - PURPOSE: To study the effect of pelvic irradiation on the level of serum prostate specific antigen (PSA). MATERIALS AND METHODS: Of 33 patients treated with pelvic irradiation to the prostate and seminal vesicles for anal and rectal cancer, 26 received 50.4 Gy or more (1. 8 Gy per fraction), and seven received 25.0 Gy (5.0 Gy per fraction). PSA levels were measured before (n = 33), during (n = 26), and after radiation therapy (n = 33). In 24 patients, follow-up (mean, 15.7 months) PSA data were obtained. Actual and pretreatment PSA levels were compared (Wilcoxon rank test). RESULTS: During the first 3 weeks in all patients, PSA levels rose steeply, culminating in a 3. 7-fold increase (P =.02). At the end of radiation therapy (7 weeks), the PSA level was no longer significantly different from the pretreatment value. In the long term, the PSA level decreased to 77% of the pretreatment value (P =.04). CONCLUSION: Irradiation of the prostate initially elevates serum PSA levels. Apparently PSA release is determined by the duration of radiation therapy, while the accumulated dose has a minor effect. In the long term, PSA production is impaired after radical radiation therapy. PSA reference concentrations should be adjusted to these reduced levels. PMID- 10831697 TI - Staging of advanced ovarian cancer: comparison of imaging modalities--report from the Radiological Diagnostic Oncology Group. AB - PURPOSE: To compare ultrasonography (US), magnetic resonance (MR) imaging, and computed tomography (CT) for diagnosing and staging advanced ovarian cancer. MATERIALS AND METHODS: US, CT, and MR imaging were performed in 280 patients. Images were read by three radiologists from each of the five hospitals. Image analysis included determination of malignancy within the peritoneum (11 sites), lymph nodes (10 sites), and hepatic parenchyma. The standard of reference was based on surgical and histopathologic findings. Statistical methods used were receiver operating characteristic (ROC) curve analysis, pairwise comparison of areas under the ROC curves (A(z)), analysis of sensitivity and specificity pairs, and assessment of agreement between the degree of suspicion and standard of reference. RESULTS: There were 118 patients with malignant tumors; 73 (62%) had stage III or IV disease. Metastases were found in the peritoneum in 70 (59%), nodes in 20 (17%), and liver in seven (6%) cases. In the peritoneum, MR imaging and CT (A(z) = 0.96 for both) were more accurate than US (A(z) = 0.86), especially in the subdiaphragmatic spaces and hepatic surfaces. MR imaging and CT were more sensitive than US (95%, 92%, and 69%, respectively) for peritoneal metastases. MR imaging was more accurate than CT for detection of lymph node metastases (A(z) = 0.76 vs 0.57, P =.04). In the liver, the A(z) values for the three modalities were 0.77-0.94. CONCLUSION: CT and MR imaging are equally accurate, and either modality can be used to stage advanced ovarian cancer. PMID- 10831698 TI - Testicular US findings after biopsy. AB - PURPOSE: To determine the ultrasonographic (US) abnormalities that may be encountered after testicular biopsy. MATERIALS AND METHODS: Testicular US studies in 33 patients (64 testes) who had undergone unilateral or bilateral testicular biopsy were retrospectively reviewed for evidence of postbiopsy changes. Biopsy had been performed in 55 testes. RESULTS: US abnormalities were found in 49 (89%) of 55 testes. These abnormalities included focal, hypoechoic, round lesions in five (9%) testes; focal hypoechoic lesions with linear margins in 27 (49%); ill defined, localized, hypoechoic areas in four (7%); peritesticular echogenic foci in 40 (73%); linear hypoechoic striations in five (9%); and focal contour defects in three (5%). CONCLUSION: A range of US findings are encountered after testicular biopsy. The US appearance of a round hypoechoic lesion seen after biopsy can overlap with that of testicular malignancy. Other findings, such as hypoechoic lesion with linear margins, peritesticular echogenic foci, linear hypoechoic striations, and testicular contour defect, can be regarded as the benign sequelae of biopsy. In a patient with a focal, nonpalpable, hypoechoic, intratesticular lesion, a history of testicular biopsy should remind the radiologist of the increased likelihood of a benign change after biopsy, in which case follow-up US may be performed. PMID- 10831699 TI - The "genetic sonogram": comparison of the index scoring system with the age adjusted US risk assessment. AB - PURPOSE: To compare two ultrasonographic (US) methods for prenatal detection of fetal Down syndrome. MATERIALS AND METHODS: Genetic amniocentesis was successfully performed in 3,303 consecutive women with high-risk pregnancies (mean gestational age, 17.1 weeks). All patients underwent a complete "genetic US" examination prospectively. Risk was assessed by using (a) various modifications of the index scoring system (ISS) and (b) the age-adjusted US risk assessment (AAURA). RESULTS: The prevalence of Down syndrome in this population was 1.6% (53 of 3,303). By using a threshold of at least 2 points to detect trisomy 21, the best ISS had a sensitivity of 45.3%, false-positive rate of 4.9%, likelihood ratio of 9.3, and positive predictive value in the high-risk population in this study of 13.3%. Lowering the threshold to 1 point increased the sensitivity to 60.4% but increased the false-positive rate to 15.8%. Adding points for age increased the sensitivity to 67.9% but increased the false positive rate to 24.3%. Results of using AAURA to detect trisomy 21 were nearly identical, with a sensitivity of 43.4% and false-positive rate of 4.9% at a 1 in 36 risk threshold and a sensitivity of 69.8% and false-positive rate of 26.1% at a 1 in 200 threshold. Trisomies 18 and 13 were detected with sensitivities of 80.0% and 100.0%, respectively, with either system. CONCLUSION: The modified ISS and AAURA are equivalent in screening for Down syndrome, with detection of approximately half of all trisomy 21 fetuses at a 5% false-positive rate. PMID- 10831700 TI - Adenomyosis: US features with histologic correlation in an in-vitro study. AB - PURPOSE: To evaluate the accuracy of ultrasonographic (US) features of adenomyosis by correlating them with histologic findings and to assess inter- and intraobserver agreement. MATERIALS AND METHODS: US was performed and videotaped in 102 consecutive hysterectomy specimens in a water bath. Videotapes were reviewed initially by two independent radiologists blinded to the clinical and histologic findings and after 1 month by one of the two; US and histologic findings were correlated. Features evaluated included diffuse abnormal echotexture of myometrium, subendometrial myometrial cysts, subendometrial echogenic nodules, subendometrial echogenic linear striations, nodular endometrial-myometrial junction, poor definition of the endometrial-myometrial junction, asymmetric thickness of the anteroposterior wall of the myometrium, and globular configuration. RESULTS: The prevalence of adenomyosis in this cohort was 29.4% (30 of 102 specimens). The mean sensitivity, specificity, negative predictive value, positive predictive value (PPV), and accuracy for the diagnosis of adenomyosis for the three reviews were 81%, 71%, 90%, 54%, and 74%, respectively. All findings evaluated, except for nodular endometrial-myometrial junction, were significantly more common in uteri with adenomyosis (P <.05). Heterogeneous myometrium reached borderline significance (P =.05). The specificities and PPVs of subendometrial striations, subendometrial echogenic nodules, and asymmetric myometrial thickness were significantly higher than those of other features (P <.05). The interobserver agreement was moderate (kappa = 0.48), and the intraobserver agreement was good (kappa = 0. 67) for the three reviews. CONCLUSION: The presence of subendometrial linear striations, subendometrial echogenic nodules, or asymmetric myometrial thickness improves the specificity and PPV of US in diagnosing adenomyosis. PMID- 10831701 TI - Power Doppler imaging: initial evaluation as a screening examination for carotid artery stenosis. AB - PURPOSE: To evaluate power Doppler imaging as a possible screening examination for carotid artery stenosis. MATERIALS AND METHODS: In the principal pilot study, a prospective, blinded comparison of power Doppler imaging with duplex Doppler imaging, the reference-standard method, was conducted in 100 consecutive patients routinely referred for carotid artery imaging at a large, private multispecialty clinic. In the validation pilot study, a prospective, blinded comparison of power Doppler imaging with digital subtraction angiography, the reference-standard method, was conducted in 20 consecutive patients routinely referred at a teaching hospital. Using conservative assumptions, the authors performed cost effectiveness analysis. RESULTS: Power Doppler imaging produced diagnostic quality images in 89% of patients. When the images of the patients with nondiagnostic examinations were regarded as positive, power Doppler imaging had an area under the receiver operating characteristic curve, A(z), of 0.87, sensitivity of 70%, and specificity of 91%. The validation study results were very similar. The cost-effectiveness of screening and, as indicated, duplex Doppler imaging as the definitive diagnostic examination and endarterectomy was $47,000 per quality-adjusted life-year. CONCLUSION: The A(z) value for power Doppler imaging compares well with that for mammography, a generally accepted screening examination, and with most other imaging examinations. Power Doppler imaging is likely to be a reasonably accurate and cost-effective screening examination for carotid artery stenosis in asymptomatic populations. PMID- 10831702 TI - Importance of thyroid abnormalities detected at US screening: a 5-year follow-up. AB - PURPOSE: To clarify the natural history and frequency of thyroid echo abnormalities in a random adult population by performing a 5-year follow-up study of subjects of a previous thyroid ultrasonographic (US) screening study. MATERIALS AND METHODS: In the original survey, 253 randomly selected adults were screened by means of thyroid US. US abnormalities were detected in 69 subjects (27%). In the follow-up study, 57 (83%) of those 69 subjects who had abnormalities were reexamined by means of thyroid US, fine-needle aspiration biopsy (FNAB), blood tests, and clinical examination. RESULTS: Of 34 individual nodules, 12 (35%) had grown. Biopsy was performed in 10 of them. Nine were benign. One was equivocal, was excised, and proved to be an adenomatous nodule. Eight nodules (24%) had diminished or disappeared. Seven new focal lesions were found in seven subjects (12%). Biopsy was performed in five of these lesions, and they were benign. At 5-year follow-up, no thyroid malignancies were detected among subjects with echo abnormalities at the primary US screening. CONCLUSION: Thyroid US abnormalities occurring in a random adult population are predominantly benign and clinically unimportant. PMID- 10831703 TI - Brain abnormalities in Gulf War syndrome: evaluation with 1H MR spectroscopy. AB - PURPOSE: To test for neuronal brain damage in the basal ganglia and brainstem in Gulf War veterans by using magnetic resonance (MR) spectroscopy. MATERIALS AND METHODS: Twenty-two Gulf War veterans with one of three factor analysis-derived syndromes (case patients); 18 well veterans matched for age, sex, and education level (control subjects); and six Gulf War veterans with syndrome 2 from a different population (replication sample) underwent long echo time (272 msec) proton (hydrogen 1) MR spectroscopy on a 4 x 2 x 2-cm voxel in the basal ganglia bilaterally and a 2 x 2 x 2-cm voxel in the pons. Syndromes 1-3 are described as "impaired cognition," "confusion-ataxia," and "central pain," respectively. RESULTS: The N-acetylaspartate-to-creatine (NAA/Cr) ratio, which reflects functional neuronal mass, was significantly lower in the basal ganglia and brainstem of Gulf War veterans with the three syndromes than in those structures of the control subjects (P =.007). The finding was corroborated in the replication sample (P =.002). Veterans with syndrome 2 (the most severe clinically) had evidence of decreased NAA/Cr in both the basal ganglia and the brainstem; those with syndrome 1, in the basal ganglia only; and those with syndrome 3, in the brainstem only. CONCLUSION: Veterans with different Gulf War syndromes have biochemical evidence of neuronal damage in different distributions in the basal ganglia and brainstem. PMID- 10831704 TI - Transfusional hemochromatosis: quantitative relation of MR imaging pituitary signal intensity reduction to hypogonadotropic hypogonadism. AB - PURPOSE: To assess the relationship between magnetic resonance (MR) imaging pituitary signal intensity reduction in patients with transfusional hemochromatosis and the clinical manifestation of hypogonadotropic hypogonadism. MATERIALS AND METHODS: Pituitary MR imaging at 0.5 T was performed in 38 consecutive patients affected by secondary hemochromatosis and in 20 healthy volunteers. Serum ferritin levels were estimated in the affected population. Twenty (53%) of the 38 patients had hypogonadotropic hypogonadism diagnosed. Pituitary-to-fat signal intensity ratios were calculated from coronal gradient echo (GRE) T2*-weighted MR images. The relationship between the quantitative reduction of the pituitary-to-fat signal intensity ratio and the clinical manifestation of pituitary dysfunction was assessed in the affected population. Signal intensity reduction in the anterior lobe of the pituitary gland was also correlated with the serum ferritin level. RESULTS: The degree of reduction of the pituitary-to-fat signal intensity ratio correlated with the presence of hypogonadotropic hypogonadism, with a sensitivity of 90%, a specificity of 89%, and an overall accuracy of 89%. In addition, the reduction of pituitary signal intensity was greater in patients with higher ferritin levels (r = -0.55, r(2) = 0.30, P <.001). CONCLUSION: The degree of signal intensity reduction, measured as the pituitary-to-fat signal intensity ratio for GRE T2*-weighted images, in patients with secondary hemochromatosis correlates with the severity of pituitary dysfunction. PMID- 10831705 TI - Multiple sclerosis: magnetization transfer MR imaging of white matter before lesion appearance on T2-weighted images. AB - PURPOSE: To determine the evolution of magnetization transfer (MT) in white matter regions before and after plaque development in patients with multiple sclerosis (MS). MATERIALS AND METHODS: In a 5-year longitudinal evaluation, 30 patients with MS underwent conventional magnetic resonance (MR) imaging, MT MR imaging, and clinical assessment. Cross-sectional data in 12 healthy subjects were also collected. Semiautomated lesion classification with use of T2-weighted MR images was used to measure the time course of the MT ratio (calculated with MR data acquired without and with MT saturation) in every voxel and to help analyze the relationship with the status of lesions depicted on T2-weighted images. RESULTS: There was a significant (P <.001) temporal decline in lesion MT ratio after lesion appearance on T2-weighted images. A significant (P <. 001) progressive decline in MT ratio was also present in voxels that later became lesions, prior to initial detection on T2-weighted images. Even 1(1/2) years prior to lesion appearance, the MT ratio (33.3%) in regions destined to become such lesions was significantly (P <.001) lower than that in both white matter in healthy subjects (41.3%) and other normal-appearing white matter in patients with MS (38.1%). CONCLUSION: The MT ratio reveals progressive focal abnormalities in MS that antedate by up to 2 years the appearance of lesions on T2-weighted MR images. PMID- 10831706 TI - Frequency and importance of transverse process fractures in the lumbar vertebrae at helical abdominal CT in patients with trauma. AB - PURPOSE: To evaluate the frequency and importance of transverse process fractures of lumbar vertebrae identified at helical computed tomography (CT) in patients with blunt abdominal trauma. MATERIALS AND METHODS: Helical abdominal CT scans in 536 consecutive patients with a history of blunt abdominal trauma were prospectively evaluated for transverse process fractures of the lumbar spine. The number and level of fractures were categorized and correlated to the retrospective and initial interpretations of the radiographs obtained at original trauma examination. Number and type of associated abdominal injuries were recorded. RESULTS: CT scans showed transverse process fractures in 39 (7.3%) patients. Seventy-nine fractures were identified (single fractures in 12 patients, multiple fractures in 27). Fractures were right-sided in 13 patients, left-sided in 24, and bilateral in two. Transverse process fractures of the L3 vertebra were most common (n = 25). Fractures were not reported in 20 (61%) of 33 initial radiographic assessments. Even at retrospective review, only 30 (57%) of 53 fractures were correctly identified. Transverse process fractures were associated with abdominal injuries in 20 (51%) patients; this association was statistically significant (P <.001). CONCLUSION: Initial conventional radiography is relatively insensitive in the detection of transverse process fractures of the lumbar spine. There is a statistically significant association between transverse process fractures and abdominal injury. PMID- 10831707 TI - Bone marrow edema pattern in osteoarthritic knees: correlation between MR imaging and histologic findings. AB - PURPOSE: To correlate magnetic resonance (MR) images of a bone marrow edema pattern with histologic findings in osteoarthritic knees. MATERIALS AND METHODS: Sixteen consecutive patients (age range, 43-79 years; mean, 67 years) referred for total knee replacement were examined with sagittal short inversion time inversion-recovery (STIR) and T1- and T2-weighted turbo spin-echo MR imaging 1-4 days before surgery. Tibial plateau abnormalities on MR images were compared quantitatively with those on histologic maps. RESULTS: The bone marrow edema pattern zone (ill-defined and hyperintense on STIR images and hypointense on T1 weighted MR images) mainly consisted of normal tissue (53% of the area was fatty marrow, 16% was intact trabeculae, and 2% was blood vessels) and a smaller proportion of several abnormalities (bone marrow necrosis [11% of area], abnormal [necrotic or remodeled] trabeculae [8%], bone marrow fibrosis [4%], bone marrow edema [4%], and bone marrow bleeding [2%]). The bone marrow edema pattern zone and the zone with a normal MR imaging appearance differed significantly in the presence of bone marrow necrosis (P =.021), bone marrow fibrosis (P =.014), and abnormal trabeculae (P =.011) but not in the prevalence of bone marrow edema (P =.069). Bone marrow edema also was found in zones with an unremarkable MR appearance (perifocal zone, 5% edema; control zone, 2% edema). CONCLUSION: A bone marrow edema pattern in osteoarthritic knees represents a number of noncharacteristic histologic abnormalities. Edema is not a major constituent of MR imaging signal intensity abnormalities in such knees. PMID- 10831708 TI - Superior labrum anterior-posterior (SLAP) tears: evaluation of three MR signs on T2-weighted images. AB - PURPOSE: To compare the sensitivity and specificity of three magnetic resonance (MR) imaging signs for the diagnosis of superior labrum anterior-posterior (SLAP) tears. MATERIALS AND METHODS: The study involved 23 consecutive patients with a type 2, 3, or 4 SLAP tear at arthroscopy and 31 age-matched control patients with an arthroscopically normal or type 1 SLAP lesion. The superior labrum was evaluated on MR images for high signal intensity extending to the articular surface in the posterior third of the labrum, an irregular or laterally curved area of high signal intensity, or two high-signal-intensity lines. RESULTS: The sensitivity, specificity, and accuracy of posterior high signal intensity for a type 2, 3, or 4 SLAP tear were 48%, 94%, and 74%, respectively, for observer 1 and 61%, 81%, and 72%, respectively, for observer 2. For laterally curved area of high signal intensity, these values were 65%, 84%, and 76%, respectively, and 56%, 84%, and 72%, respectively. For two high-signal-intensity lines, these values were 17%, 94%, and 61%, respectively, and 13%, 94%, and 59%, respectively. For the presence of either posterior or laterally curved high signal intensity, the sensitivity was 65% for both observers, whereas the specificity was 84% for observer 1 and 74% for observer 2. The kappa values for interobserver agreement were 0.60 for posterior high signal intensity and 0.58 for laterally curved high signal intensity. CONCLUSION: Laterally curved and posterior high signal intensities are specific signs for distinguishing a SLAP tear from a normal variant superior sublabral recess. PMID- 10831709 TI - Articular cartilage defects: in vitro evaluation of accuracy and interobserver reliability for detection and grading with US. AB - PURPOSE: To determine the accuracy and reliability of detecting and grading articular cartilage defects in porcine and human knees by using ultrasonography (US). MATERIALS AND METHODS: US was used to evaluate 175 porcine and 16 human knee surfaces with a linear 5-12-MHz transducer. Porcine defects of varying diameter and depth were surgically created. Each porcine surface was independently assessed in blinded fashion by two radiologists for the presence and severity of defects. Accuracy of detection, interobserver reliability, and concordance between US and surgical grades were determined. Human specimens were retrieved from knees of patients who underwent joint arthroplasty. Defects in human knees detected with US were correlated with defects seen at direct surface visualization. RESULTS: Sensitivities for detection of porcine defects were 94% and 93% for readers 1 and 2, respectively; specificities were 90% and 77%, respectively; positive predictive values were 98% and 95%, respectively; and negative predictive values were 78% and 73%, respectively. Interobserver agreement was high (weighted kappa = 0.80), and concordance between US and surgical grades for both readers was high (weighted kappa = 0.90 and 0.78). In human cartilage, the distribution of cartilage denudation determined at US was the same as that determined at direct visualization. CONCLUSION: High-frequency US was accurate and reliable for detection and grading of knee articular cartilage defects. PMID- 10831710 TI - Hepatic colon cancer metastases in mice: dynamic in vivo correlation with hypoechoic rims visible at US. AB - PURPOSE: To use videomicroscopy of tumor-bearing livers of live mice to depict tumors directly to determine the exact nature of rims seen on corresponding ultrasonographic (US) scans. MATERIALS AND METHODS: Seventy-six hepatic colorectal cancer metastases were studied in exteriorized livers of 18 mice by using intravital microscopy, US, and histologic examination of the same tumors. RESULTS: Hypoechoic rims correlated with distended sinusoidal spaces in vivo. These spaces surrounded only locally invasive tumors (mean diameter, 0.85 mm) that had obstructed the supplying terminal portal venules. These spaces, containing adherent leukocytes and tumor cells, gave rise to new tumor vasculature. Results of histologic examination of rims (portal inflammation, congested or compressed sinusoids, cell atrophy) correlated with leukocyte endothelial adherence, occluded sinusoids, and new vessel formation in vivo. CONCLUSION: Unlike results from previous studies, dynamic in vivo observations of peritumoral rims demonstrated distended sinusoidal spaces giving rise to new tumor-penetrating vessels. These sinusoids arose around locally invasive tumors and were associated with more advanced intrahepatic disease. These dynamic observations provide a pathophysiologic explanation for previous histologic correlates of peritumoral rims. PMID- 10831711 TI - Human fetal lung fibroblasts: in vitro study of repetitive magnetic field exposure at 0.2, 1.0, and 1.5 T. AB - PURPOSE: To evaluate whether repetitive exposure to magnetic fields of 0.2, 1.0, and 1.5 T affect the growth of human fetal lung fibroblasts (HFLFs). MATERIALS AND METHODS: Cultured HFLFs were exposed to static magnetic fields of 0.2, 1.0, and 1.5 T for 1 h/d for 5 consecutive days. Control groups were kept under identical environmental conditions, apart from the magnetic field, during the experiments. Cell cycle analysis for synchronously and nonsynchronously growing cells was performed. Population doublings (PDs) were calculated. To rule out midterm effects, proliferation kinetics of the cells were analyzed for 21 days. RESULTS: Cell cycle analysis of synchronized and nonsynchronized cells did not reveal statistically significant differences between the exposed and control cells. The PDs did not indicate any growth modulation during exposure. Proliferation kinetics did not provide any hint of midterm growth modulation effects of repetitive magnetic field exposure. CONCLUSION: Repetitive magnetic field exposure does not exert any growth-modulating effect on overall cell growth and cell cycle distribution of cultured HFLFs. Midterm effects due to magnetic field exposure were not found. PMID- 10831712 TI - Irreversibly damaged myocardium at MR imaging with a necrotic tissue-specific contrast agent in a cat model. AB - PURPOSE: To investigate the capability of a necrosis-avid magnetic resonance (MR) contrast agent, bis-gadolinium mesoporphyrins, for assessment of irreversibly damaged myocardium and to evaluate the time course of signal enhancement in the reperfused myocardium. MATERIALS AND METHODS: Nine cats were subjected to 90 minutes of occlusion of the left anterior descending coronary artery followed by 90 minutes of reperfusion. Contrast material-enhanced T1-weighted spin-echo images were obtained for 12 hours in five cats and 6 hours in four cats. Pathologic examinations of the resected specimens were performed with 2'3'5 triphenyl tetrazolium chloride (TTC) histochemical staining and electron microscopy. The size of enhanced area on MR images was compared with that of irreversibly damaged myocardium with TTC staining. The time course of signal enhancement was evaluated. RESULTS: The size of enhanced area on MR images was well correlated with that of irreversibly damaged myocardium with TTC staining. Maximum enhancement occurred 1-3 hours after administration of the contrast material, with mean enhancement of 171% that of normal myocardium. Electron microscopic examinations showed severe myocardial damage in the irreversibly damaged myocardium but only mild edematous changes in the reversibly damaged myocardium. CONCLUSION: MR images enhanced with bis-gadolinium mesoporphyrins provide accurate sizing of irreversibly damaged myocardium with a strong and persistent signal enhancement in the reperfused myocardium. PMID- 10831713 TI - MR imaging and cardiac pacemakers: in-vitro evaluation and in-vivo studies in 51 patients at 0.5 T. AB - PURPOSE: To evaluate the safety and feasibility of magnetic resonance (MR) imaging at 0.5 T in patients with implanted cardiac pacemakers. MATERIALS AND METHODS: Twenty-one models of pacemakers and 44 pacemaker electrodes were exposed to in vitro MR imaging with continuous registration of pacemaker output and temperature at the lead tip. Prior to MR imaging examination, pacemakers were programmed to an asynchronous mode (A00, V00, or D00). Pacemakers were examined before and after MR imaging. Forty-four patients with implanted pacemakers underwent 51 MR imaging examinations under cardiologic surveillance, continuous electrocardiography, pulse oximetry, and capnographic monitoring. RESULTS: MR imaging was safely performed in all patients. None of the pacemakers displayed a pacing dysfunction at MR imaging. No changes occurred in the programmed parameters in any device tested in vivo or in vitro. Maximum increases in the temperature at the lead tips were 8.90 degrees C at a specific absorption rate (SAR) of 0.6 W/kg and 23.50 degrees C under a worst-case radio-frequency (RF) heating condition with an SAR of 1.3 W/kg. CONCLUSION: MR imaging at 0.5 T can be safely performed in patients with implanted pacemakers in carefully selected clinical circumstances when appropriate strategies (programming to an asynchronous mode, adequate monitoring techniques, limited RF exposure) are used. PMID- 10831714 TI - Q fever pneumonia: CT findings. AB - PURPOSE: To evaluate the computed tomographic (CT) features of Q fever pneumonia. MATERIALS AND METHODS: The authors retrospectively reviewed the chest radiographs and CT scans obtained in 12 patients, who were selected on the basis of chest CT availability from a group of patients with a definite diagnosis of acute Q fever infection during an 8.5-year period. RESULTS: In all cases, CT depicted lesions indicative of airspace involvement, which was expressed as lobar (n = 3), segmental (n = 3), patchy (n = 3), or a combination of these patterns (n = 3). Involvement of more than one lobe was observed in seven (58%) patients. In one patient with multiple patchy areas of consolidation, nodular lesions with a vascular connection and a halo of ground-glass opacity, which were suggestive of an angioinvasive process, were demonstrated. In addition, CT performed in a patient with acute Coxiella burnetii infection who abused alcohol revealed necrotizing pneumonia. Pleural effusions were seen at both CT and radiography in three patients, and mild lymph node enlargement in isolated regions was seen at CT in four patients. Chest radiography was less accurate than CT in the detection of segmental and patchy areas of consolidation. CONCLUSION: The typical CT findings of Q fever pneumonia consisted mainly of multilobar airspace consolidation. A nodular pattern accompanied by a halo of ground-glass opacification and vessel connection, and necrotizing pneumonia in the setting of impaired immunity were less frequent. PMID- 10831715 TI - The panda sign. PMID- 10831716 TI - Non-small cell lung cancer: FDG PET for nodal staging in patients with stage I disease. AB - PURPOSE: To determine the accuracy of 2-[fluorine-18]fluoro-2-deoxy-D-glucose (FDG) positron emission tomography (PET) in the evaluation of regional lymph nodes in patients with stage I non-small cell lung cancer (NSCLC). MATERIALS AND METHODS: Imaging and clinical findings obtained during 5 years in 84 patients (mean age, 66 years) were reviewed. Patients had thoracic computed tomographic findings of stage I NSCLC, an FDG PET study, and histopathologic proof of lung cancer. At the time of diagnosis, disease stage was assigned on the basis of FDG PET results and was compared with the histopathologic stage to determine the accuracy of PET. RESULTS: When PET stage was compared with histopathologic stage, the disease in 72 (86%) patients was accurately staged with PET, understaged in two (2%), and overstaged in 10 (12%). The overall sensitivity, specificity, and positive and negative predictive values for PET of regional lymph nodal metastases were 82%, 86%, 47%, and 97%, respectively. CONCLUSION: FDG PET enables accurate staging of regional lymph node disease in patients with stage I NSCLC. A negative PET scan in these patients suggests that mediastinoscopy is unnecessary and that these patients can proceed directly to thoracotomy. PMID- 10831717 TI - Results of a nationwide survey of chest radiography: comparison with results of a previous study. AB - PURPOSE: To provide public health information by means of measurement of the radiation exposures that patients undergoing chest radiography would receive and to compare the results with those of a similar previous survey. MATERIALS AND METHODS: Surveyed facilities were randomly selected from each state. Patient exposure was evaluated along with film processing, half-value layer, and image quality. Additional information obtained concerned type of equipment, facility work load, radiographic technique, screen-film system, and grid type. RESULTS: Mean entrance air kerma in all facilities was 141 microGy (16.1 mR). Mean kilovoltage in all facilities was 101 kV. In 1994, 140 (90%) of 156 hospitals (vs 71% in 1984) and 92 (58%; nearly double the percentage in 1984) of 159 nonhospital sites were using grids. Scoring with the imaging test tool resulted in a mean spatial resolution of 2.3 cycles per millimeter, and a mean low contrast sensitivity of about 3%. Two hundred fifty-three (80%) of 315 facilities surveyed were processing film at minimum acceptable performance levels. CONCLUSION: Mean entrance air kerma for all facilities did not substantially change. Although increased grid usage would lead to the expectation of higher measured exposures, this was offset by an increase in the use of faster screen film combinations. PMID- 10831718 TI - Reduced frequency of sedation of young children with multisection helical CT. AB - PURPOSE: To determine whether the use of multisection helical computed tomography (CT) can decrease the need for sedation compared with single-section helical CT. MATERIALS AND METHODS: The number of children who required sedation to undergo body CT with a multisection helical scanner was recorded. The authors noted the type of examination and whether contrast material was used. The children were categorized according to age (< or = 17 years, < or = 6 years, < or = 1 year). RESULTS: In 219 CT examinations, only three children required sedation (1.4%). The sedation rate was 3% (three of 90) for children aged 6 years or younger and 8% (three of 37) for those aged 1 year or younger. Examinations were of the chest, abdomen, and pelvis in 68 patients, of the abdomen and pelvis in 112, and of the chest alone in 39. Contrast material was intravenously administered in 186 (85%) examinations. All scans were of diagnostic quality. CONCLUSION: The rate of sedation was reduced threefold with multisection helical CT compared with standard helical CT, and the need for sedation was eliminated in some age groups. PMID- 10831719 TI - Gynecologic brachytherapy: digital fluoroscopy for placement verification and treatment planning. AB - The authors evaluated the feasibility of using digital fluoroscopic images for device placement verification and dosimetric planning for gynecologic brachytherapy. Adequate images were obtained rapidly, and the limited pincushion distortion on digital fluoroscopic images produced negligible variations in brachytherapy dose calculations compared with those calculated with standard radiographs. Intraoperative digital fluoroscopy can facilitate both placement verification and dosimetric planning for gynecologic brachytherapy. PMID- 10831720 TI - Autocorrection in MR imaging: adaptive motion correction without navigator echoes. AB - A technique for automatic retrospective correction of motion artifacts on magnetic resonance (MR) images was developed that uses only the raw (complex) data from the MR imager and requires no knowledge of patient motion during the acquisition. The algorithm was tested on coronal images of the rotator cuff in a series of 144 patients, and the improvements in image quality were similar to those achieved with navigator echoes. The results demonstrate that autocorrection can significantly reduce motion artifacts in a technically demanding MR imaging application. PMID- 10831721 TI - Normal lumbar vertebrae: anatomic, age, and sex variance in subjects at proton MR spectroscopy--initial experience. AB - Fifty-seven subjects underwent proton magnetic resonance (MR) spectroscopy of the second lumbar vertebra to evaluate single-voxel and multivoxel techniques. Measurements included lipid-to-water ratios, lipid fractions, and line width. These data provide information about vertebral fat content. There was an age dependent linear increase in fat content and sex dependence. A higher fat concentration was found in men. The observed spectra provide a basis for future study to determine clinical utility of vertebral proton MR spectroscopy. PMID- 10831722 TI - Intraoperative US versus intraoperative MR imaging for guidance during intracranial neurosurgery. PMID- 10831723 TI - Diagnostic accuracy of thin-section CT in idiopathic interstitial pneumonia. PMID- 10831724 TI - Dynamic spiral MR mammography. PMID- 10831725 TI - Sentinel lymph node biopsy in breast carcinoma. PMID- 10831727 TI - Abstracts of current literature PMID- 10831726 TI - Meningeal enhancement on fast FLAIR images. PMID- 10831728 TI - Asbestos exposure and laryngeal cancer. AB - OBJECTIVE: To carry out a systematic review of the evidence relating asbestos exposure to the risk of laryngeal cancer. METHOD: All identified studies of asbestos workers providing data on laryngeal disease were reviewed, together with studies of laryngeal cancers giving epidemiological or experimental evidence of associated exposures. RESULTS: Confounding due to smoking and alcohol intake, and to a lesser extent diet and socio-economic factors, creates a major difficulty over the identification of any asbestos or other occupational effect. Not only are smoking and alcohol independently associated with large increases in relative risk (RR) of laryngeal cancer, but also have a synergistic effect with each other. Few of the studies provide details of either habit. Among 24 prospective studies for which a standardized mortality ratio (SMR) was available, nine had an SMR at or below unity, and among a further 11 without an SMR for comparison, in only one was there a clear excess risk. In 17 retrospective studies, only two showed a significantly increased RR. Evidence from animal experiments, studies of associations with pleural plaques, and autopsy findings also appear negative or inconclusive. CONCLUSION: The evidence does not indicate that asbestos exposure increases the RR of laryngeal cancer. PMID- 10831729 TI - Estimating factors to convert Chinese 'Total Dust' measurements to ACGIH respirable concentrations in metal mines and pottery industries. AB - Historical data on the dust exposures of Chinese workers in metal mines (iron/copper, tin, tungsten) and pottery industries are being used in an ongoing joint Chinese/United States epidemiological study to investigate the exposure response relationship for the development of silicosis, lung cancer, and other diseases. The historical data include 'total dust' concentrations determined by a Chinese method. Information about particle size distribution and the chemical and mineralogical content of airborne particles is generally not available. In addition, the historical Chinese sampling strategy is different from a typical American eight-hour time-weighted average (TWA) sampling strategy, because the Chinese samples were collected for approximately 15 minutes during production so the sample could be compared to their maximum allowable concentration (MAC) standard. Therefore, in order to assess American respirable dust exposure standards in light of the Chinese experience, factors are needed to convert historical Chinese total dust concentrations to respirable dust concentrations. As a part of the joint study to estimate the conversion factors, airborne dust samples were collected in 20 metal mines and 9 pottery factories in China during 1988 and 1989 using three different samplers: 10mm nylon cyclones, multi-stage 'cassette' impactors, and the traditional Chinese total dust samplers. More than 100 samples were collected and analysed for each of the three samplers. The study yielded two different estimates of the conversion factor from the Chinese total dust concentrations (measured during production processes) to respirable dust concentrations. The multivariate analysis of variance (MANOVA) reveals that, with a fixed sampling/analysis method, conversion factors were not statistically different among the different job titles within each industry. It also indicates that conversion factors among the industries were not statistically different. However, the two estimates consistently showed that conversion factors were the lowest in the pottery industry. Average conversion factors were then calculated for each of the estimates across the industries studied. A pooled mean conversion factor, 0.25+/-0.04, was then derived for all the job titles and industries. Respirable dust levels were estimated from the historical 'total dust' concentrations collected between 1952 and 1992 by adopting the American standard. PMID- 10831730 TI - Inflammatory potential of dust from waste handling facilities measured as IL-8 secretion from lung epithelial cells in vitro. AB - OBJECTIVES: Organic dust contains several different components which may cause pulmonary effects, and many health problems have been associated with the collection and recycling of organic waste. It is often difficult to obtain a precise measurement of the exposure to each component in dust, and organic dust samples obtained from different workplaces may vary profoundly in composition. The aim of this study was to evaluate the inflammatory potential of dust from different waste handling plants. Furthermore, we set out to investigate the role of endotoxin in the inflammatory potential of dust. METHODS: Dust samples were obtained from four incineration plants, three samples from a plant sorting household waste, five paper-sorting plants, two mail centres, four bottle-sorting plants, and two combined paper-sorting and composting plants. The samples were tested in a bioassay with the lung epithelial cell line A549. Cells were stimulated for 24h with dust samples at six concentrations, and subsequently the interleukin 8 (IL-8) secretion into the growth medium was measured. The initial slope of the dose response curves was used to calculate the potency factor (PF) of the dust samples, and correction against positive control samples was used to reduce day-to-day variation. The concentration of endotoxin in the dust samples was measured by the limulus amebocyte lysate (LAL) assay. RESULTS: The inflammatory potential of the dust samples for dust from the paper- and mail sorting plants showed a significantly lower PF as compared with dust from the plants handling mixed household waste. A significantly lower PF for the dust samples from the bottle-sorting plants (excluding one outlier plant) compared with dust from the plants handling mixed household waste was also found. No correlation was observed between the PF and the concentration of endotoxin in the samples. CONCLUSION: The PFs obtained seem to reflect the material handled, with mixed household waste generating organic dust with the highest inflammatory potentials. PMID- 10831731 TI - Trends in levels of inhalable dust exposure, exceedance and overexposure in the European carbon black manufacturing industry. AB - In an attempt to investigate the relationship between exposure to carbon black and respiratory morbidity, a study of the complete carbon black manufacturing industry in Western Europe was commissioned. As part of this study, a large number of personal inhalable (n=8015) dust exposure measurements was taken during three phases of data collection between 1987 and 1995. Repeated measurements on the same worker were taken in the last two phases, which enabled the estimation of the within- and between-worker components of variance. Simultaneously, the fixed effects of phase and factory were estimated using mixed-effects analysis of variance. The results show that the personal inhalable dust exposure has reduced significantly since the first phase of the study. In addition, the interaction term between phase and factory was significant in most job categories, confirming that the reduction of exposure was not equal across all factories. When all factories were considered together, the probability that the mean exposure of a randomly selected worker (overexposure) or the probability that the exposure on a randomly selected day for a randomly selected worker (exceedance) was higher than 3.5mg m(-3), was 10% or less for all job categories in the last phase. However, when the factories were considered separately, it appeared that the probability of overexposure or exceedance was in excess of 10% for the job categories 'Fitter/Welder', 'Warehouseman' and 'Site crew' for a number of factories. Therefore, even though exposure levels of inhalable dust have dropped considerably across the whole carbon black manufacturing industry in Western Europe, further reductions in exposure levels are required in these areas to make sure that the probability of overexposure and exceedance falls below a level of 10%. PMID- 10831732 TI - Effects of personal exposures on pulmonary function and work-related symptoms among sawmill workers. AB - Three green mills and two dry mills were studied for personal exposure to wood dust and biohazards associated with wood dust and their correlation to lung function and work-related symptoms among sawmill workers. The levels of exposure to endotoxin, (1-->3)-beta-D-glucan, bacteria and fungi were high in green mills compared with dry mills. Compared with dry mill workers, green mill workers had significantly high prevalence of regular cough, chronic bronchitis, regular blocked nose, regular sneezing, sinus problems, flu-like symptoms, and eye and throat irritation. Significant positive correlations were found among endotoxin and Gram (-)ve bacteria, (1-->3)-beta-D-glucan and fungi, and endotoxin and (1- >3)-beta-D-glucan exposure levels. Significant dose-response relationships were found for personal exposures and lung function, and lung function and work related respiratory symptoms. The significant correlations found for respirable fractions show that not only inhalable but also respirable fractions are important in determining potential health effects of exposure to wood dust. The management and employees of the sawmilling industry should be educated on the potential health effects of wood dust. Wood dust should be controlled at the source. PMID- 10831733 TI - A simple method for fume cupboard performance assessment. AB - The performance of a fume cupboard is determined by a complex interaction of factors which are time consuming and expensive to determine. This paper describes a simple and practical means of ranking, and assessing fume cupboard installations that can help to discharge managerial responsibility for a 'safe' environment. The method also gives an economically viable and technically defensible system for assessing fume cupboard performance as part of upgrading exercises or performance audits. The assessment strategy uses flow visualisation techniques and measurements of inflow air velocity as well as overall condition evaluation to rank performance and identify poor performing cupboards. The method has been used to carry out a condition and performance survey of 199 fume cupboards, both aerodynamic and box-type designs, in an academic institution. The results of this survey are presented which not only highlight performance characteristics but also provide insights into user attitudes and knowledge of fume cupboard operation and performance. It is suggested that surveys such as this could be helpful in training programmes for laboratory workers to enable them to optimise the use of fume cupboards. PMID- 10831734 TI - Retrospective exposure assessment for benzene in the Australian petroleum industry. AB - An excess of lympho-haematopoietic (LH) cancers has been identified in the Australian petroleum industry through the Health Watch surveillance programme. A nested case-control study is being conducted to investigate this excess. This paper describes the methods used to provide quantitative estimates of benzene exposure for each of the subjects in the case-control study. Job histories were compiled for each subject from interviews and company employment records. Site visits and telephone interviews were used to identify the tasks included in each job title. Details about the tasks such as their frequency, the technology in use and about changes that had taken place over the years were also gathered. Exposure dated back to the late 1940s for a few subjects. Collaborating petroleum companies provided recent benzene exposure monitoring data. These were used to generate Base Estimates of exposure for each task, augmented with data from the literature where necessary. Past exposures were estimated from the Base Estimates by means of an exposure algorithm. The modifying effects of technological changes and changes to the product were used in the algorithm. The algorithm was then computed to give, for each job, for each subject, an estimate of average benzene exposure in ppm in the workplace atmosphere (Workplace Estimate). This value was multiplied by the years for which the job was held and these values summed to give an estimate of Cumulative Estimate of benzene in ppm-years. The occupational hygienists performing the exposure assessment did so without knowledge of the case or control status of subjects. Overall exposures to benzene in the Australian petroleum industry were low, and virtually all activities and jobs were below a time-weighted average of 5 ppm. Exposures in terminals were generally higher than at refineries. Exposures in upstream areas were extremely low. Estimates of Cumulative Estimate to benzene ranged from 0.005 to 50.9 ppm years. PMID- 10831735 TI - Factors affecting mortality and morbidity in patients with abdominal gunshot wounds. AB - Risk factors that may independently predict mortality and morbidity in patients with abdominal gunshot wounds have not been fully elucidated. We prospectively studied the effects of 12 potential risk factors on mortality and morbidity in 82 patients with abdominal gunshot wounds who required laparotomy. Univariate analysis of these factors revealed that shock on admission, presence of penetrating colon injury and number of intra-abdominal organs injured (NOI)>2 were associated with greater than threefold increased incidence of death (p<0.05). Penetrating abdominal trauma index (PATI) score>15 was associated with twentyfold increased incidence of death (P<0.0001). Multivariate analysis showed that only PATI (P=0.001), number of postoperative complications per patient (N(comp)) (P=0.023) and presence of shock on admission (P=0. 028) were independently significant in predicting mortality. PATI was the only risk factor that independently predicted the development of postoperative infectious complications and N(comp) (P<0.0001). The type of gun used was not a significant risk factor (P>0.05). The 15 (18.3%) non-survivors were significantly older than survivors (P=0.02), had longer operations (P=0.004) and their NOI, PATI and N(comp) were significantly higher (P<0.001). The uniformly prolonged injury to surgery time in all patients contributed to the high incidence of infectious complications (62.2%) and mortality. PATI score was the most important factor found to be independently associated with mortality and morbidity in our subset of patients with prolonged injury to surgery time and high rate of colon injury. PMID- 10831736 TI - Behaviour of an artery enclosed within a normal or hypertrophic callus of a long bone fracture. AB - The behaviour of the brachial artery enclosed in a hypertrophic or a normal callus was investigated in experimentally produced fractures of the humerus in 22 dogs. The brachial artery was displaced in a bony groove created at the fractured ends of the bone. The fracture was immobilized with a metal plate and four screws. The progress of the callus formation was studied and the patency of the artery was evaluated. In 15 out of the 22 animals a medium-sized or hypertrophic callus had developed that engulfed the brachial artery without obstructing its lumen and blood flow. In five dogs the fracture site was infected and the resultant osteomyelitis obstructed the artery. In the remaining two dogs the arterial lumen was extremely narrowed, due to breaking of the plate and formation of pseudarthrosis in one and injury of the artery in the other. Unless complicated by infection resulting in vascular occlusion, callus at the fracture site may engulf an artery without interference in its patency and blood flow. The possible involvement of a functioning artery within a callus or a mass of heterotopic ossification (myositis ossificans) should be kept in mind during surgical treatment of old fractures, hypertrophic callus with pseudarthrosis or extensive heterotopic ossification. PMID- 10831737 TI - Complications of tibial shaft soccer fractures. AB - A retrospective study of AO type 42 tibial diaphyseal fractures that presented to a teaching hospital over a 54 month period was made to identify the proportion sustained whilst playing soccer, determine their characteristics and report treatment and outcome. Sport accounts for 73/329 (22.1%) of these fractures and soccer 58/73 (79. 5%) of these. All patients were male with mean age of 24.3 years (range 8-48). Fifty-four fractures were closed and 93.1% (54/58) were situated in the middle third or at the junction of the middle and distal thirds of the diaphysis. Fifty-six (96.6%) had simple or wedge patterns and 45 (77.6%) were right sided. Forty-four (76.2%) were treated non-operatively in plaster, 12 (20.3%) by intramedullary nails and two (3.4%) with external fixators. Two patients were lost to follow-up and the remaining 56 fractures united at a mean of 6.5 months. There were 21 complications in 19/56 (33.9%) patients which included 8/56 (14.3%) delayed/non-unions requiring surgery. There was a significantly higher complication rate for operated fractures (p<0.005) but no significant link to AO fracture type. Thus we cannot assume that treatment of these common fractures is without risk, especially if they are treated operatively. PMID- 10831738 TI - Contralateral hip fractures - can predisposing factors be determined? AB - A case control study was carried out in the Orthopaedic Department of Bradford Royal Infirmary in an attempt to see if certain medical conditions, which can affect balance and stability, are more common in those who sustain a second proximal femoral fracture. Medical conditions included in the study were: late effects of cerebro-vascular accident, blindness, syncope and collapse, alcoholism, Alzheimer's disease, epilepsy, Parkinsonism, ischaemic heart disease and senile dementia. The study group comprised 53 patients admitted to hospital between 1992 and 1998 with two separate proximal femoral fractures each on a different side. The control group comprised 530 patients selected from a general pool of 2080 proximal femoral fracture patients admitted to hospital during the same period. The control group patients were matched to the study group for age, sex, and time of occurrence of the first fracture. Results show significantly higher association of late effects of cerebro-vascular accident, blindness, syncope and collapse, and Alzheimer's disease with subsequent contralateral proximal femoral fractures. This study supports a causal relationship between the above medical conditions and subsequent contralateral proximal femoral fractures. It may therefore be possible to identify patients who are at risk of returning with a second fracture. PMID- 10831739 TI - Eponyms - are they relevant? AB - This study was performed to investigate the value of eponyms of fractures amongst orthopaedic/casualty trainees. Our results show eponyms are not favoured. PMID- 10831740 TI - K-wire position in tension band wiring of the olecranon - a comparison of two techniques. AB - Tension band wiring is a recognised standard treatment for olecranon fractures. We studied the effect of K-wire position on backing out of the wire in a group of 80 patients with closed transverse olecranon fractures with a minimum follow-up time of 9 months. The rate of wires backing out as seen on X-ray was three times greater in patients who had K-wires passed down the long axis of the ulna rather than across the anterior cortex as recommended by the AO group. There was a corresponding higher rate of local complications in these patients. 42% of this group had to have the metal removed compared with 11.4% of the transcortical group. We compared the biomechanical properties of both K-wires positions in a human cadaveric model. The maximum pull-out strength for each configuration was recorded in 20 elbow joints. The average maximum pullout strength for the intramedullary wires was 56.3 N (range 27. 7-95.6 N) and 122.7 N for the transcortical wires (range 56.7-201.2). The results of both the clinical study and biomechanical data support the routine use of transcortical placement of K wires in tension-band wiring of transverse olecranon fractures. PMID- 10831741 TI - A simple method for removal of a broken intramedullary nail. AB - The removal of broken implanted intramedullary nails secondary to re-fracture or non-union is challenging. In 12 cases a simple and safe method has been used to remove broken implants. PMID- 10831742 TI - Analgesia requirements following hip fracture in the cognitively impaired. AB - The analgesia received in the 1st week of treatment by 100 consecutive acute hip fracture patients (88 female) was prospectively studied. A modified mental test was performed on all patients prior to surgery. Patients with cognitive impairment (modified mental score <5) received only 74% of the paracetamol, 43% of the non-steroidal anti-inflammatory drugs (NSAIDs), 52% of the opioid and 64% of the morphine that cognitively intact patients received. These differences were significant for paracetamol (Mann-Whitney test; p=0.001) and opioid (Mann-Whitney test; p=0. 0012) but not for NSAIDs or morphine. Either hip fracture patients with a reduced mental score perceive less pain than their more cognisant peers, or the treating clinicians perceive their pain as less, or both. PMID- 10831743 TI - Biomechanics of femoral interlocking nails. AB - Today there is a variety of different interlocking intramedullary nail designs available for the femur. We compared different nail types in the bone implant complex (BIC) of four unreamed solid nails and a slotted reamed nail with simulated comminuted mid shaft fractures to see if there are major differences in stiffness for axial load, bending and torsion. The fractures were simulated by a 2 cm defect osteotomy in paired human cadaver femora. Each bone was tested intact in a universal testing machine, osteotomy and osteosynthesis were performed, and the BIC was tested. Relative stiffness was calculated. In torque testing the unslotted solid nail showed significantly more stiffness (0.6-1.8 Nm/degrees) compared to the slotted nail (0.2 Nm/degrees). Compared to intact bone (6.9 Nm/degrees), both groups of nails were significantly less stiff (relative stiffness 2-20%). In axial load and bending testing the large diameter unreamed nail showed significantly higher stiffness (32-68%). This study shows that stiffness of bone implant complex in interlocking femoral nails is more dependent on nail profile than on the pressfit of nails in the medullary canal. PMID- 10831744 TI - Initial experience with the Forte plate for dorsally displaced distal radius fractures. AB - Open reduction and plate osteosynthesis is occasionally indicated for dorsally displaced distal radius fractures. We reviewed our medium term results with the Forte plate, one of the recently introduced purpose-made implants.Twenty-five patients operated on during the first year were reviewed 19 (12-24) months after surgery. Median age at operation was 53 (28-80) years. There were seven high energy and eighteen low energy injuries. Fourteen fractures extended into the radiocarpal joint.Three patients had a poor clinical result and were re-operated on before review with an arthrodesis, ulnar shortening, or Sauve-Kapandji operation. The remainder had six excellent, twelve good, and four fair results. Irritation of the extensor tendons was a minor problem. Initial radiological correction of deformity was satisfactory, but increased volar angulation of the distal radius was seen at follow up in twenty patients - by more than 10 degrees in nine. Seven patients had 20-30 degrees volar tilt at final review and tended to have a poorer clinical result than other patients. In our patients use of the Forte plate seems to have given satisfactory clinical results, but the increase in volar tilt after surgery is a cause for concern. PMID- 10831745 TI - Preoperative templating of tibial nails - is it worthwhile? PMID- 10831746 TI - The clinical performance of a small diameter tibial nailing system with a mechanical distal aiming device. AB - We present the clinical and radiographic results of a prospective study with the Orthofix tibial nailing system. The ease and safety of distal locking with the use of an improved targeting system was also evaluated. Sixty fresh tibial fractures in 60 patients with a mean age of 37.3 years (range 17-73 years) were treated. Eighteen of the fractures were grade I open fractures. All operations were performed in a conventional operating theatre on a simple transparent operating table, with reduction of the fracture performed under manual traction and manipulation of the fracture site. Hand reaming was then performed to ensure, where possible, the insertion of a nail of at least 9 mm in diameter. Fracture healing was observed at a mean of 17 weeks (12-28 weeks). No tibial non-unions occurred in our series, and only three fractures, two segmental and one severely comminuted, showed delayed union. No infection, either superficial or deep, was found and no cardio-pulmonary complications were recorded. Following surgery, all patients gained a full range of pain-free movement of the ankle and knee joints and only six patients (10%) complained of mild anterior knee pain. All patients returned to their previous jobs one month after fracture healing had been confirmed clinically and radiographically. Following nailing, no deviation from normal tibial alignment was detected. No mechanical failure of either the nails or the locking screws was recorded. The mean duration of operation (skin to skin) was 30 min (range 20-45 min) and the mean total theatre time was 55 min (range 40 75 min). The mean total intensification time was 5 s. In total, 120 distal locking screws were inserted using the external targeting device. All attempts at distal locking except five (4.2%) were successful with two failures in the same patient being a result of inappropriate use of the system. We conclude that this nailing system is clinically effective and that distal locking can be performed easily, without exposure to radiation. PMID- 10831747 TI - Wound complications following operative fixation of calcaneal fractures. AB - The aim of our study was to find the incidence of wound complications following operative fixation of fractured calcanea and identify the risk factors contributing to them. We retrospectively reviewed the results of operative treatment of 33 calcaneal fractures in 30 patients over a 4-year period. We report an overall wound complication rate of 18.1%. Wound infection, haematoma, dehiscence and heel necrosis were noted in our series with or without underlying osteomyelitis. We identified the following as risk factors to the causation of post-operative wound complications: (a) fall of more than 3.4 m (p<0.005); (b) surgery within 7 days (p<0.05); (c) operating time in excess of 2 h (p<0.05); (d) tourniquet time in excess of 1.5 h (p<0.001). We recommend careful attention to these factors in treating calcaneal fractures. PMID- 10831748 TI - Fracture-dislocation of the first metatarso-phalangeal joint. PMID- 10831749 TI - Bilateral simultaneous atraumatic quadriceps tendon ruptures associated with 'pseudogout'. PMID- 10831750 TI - Irreducible anterior shoulder dislocation: the greater tuberosity Hill-Sachs lesion. PMID- 10831751 TI - Purtscher's retinopathy. PMID- 10831752 TI - Angle grinder injuries. PMID- 10831753 TI - Simulating the response of a standing operator to vibration stress by means of a biomechanical model. AB - Under vibration stress the compressive forces transmitted in the joints of a standing operator are composed of nearly static and oscillating force parts. Because these forces can hardly be measured they were assessed by means of a biomechanical model. In the model, 27 rigid bodies with 103 degrees of freedom represent the segments of the human body. 106 force elements imitate the muscles of the trunk and the legs. At first, the model parameter were varied so that for the simulated sitting posture the model fits the seat-to-head transmissibility given in the literature and in ISO/CD 5982. For the standing posture, the transfer functions between the ground acceleration and the oscillating forces in the ankle, the knee, the hip, and the motion segment L3-L4 were computed. According to the moduli of these functions the forces in the ankles are higher than those in the knees or the hips and they nearly come up to the forces in the lumbar spine. Further the results of the simulation indicate that under equal vibration stress in the standing and the sitting posture the differences between the compressive forces in the lumbar spine are small. PMID- 10831754 TI - Foot displacement but not velocity predicts the outcome of a slip induced in young subjects while walking. AB - The purpose of the present study was to induce slips in healthy subjects as a means to determine if recovery from an induced slip is possible under conditions in which the displacements and velocities of the slipping foot exceed the generally accepted limits of 10cm and 50cm/s, respectively, and to determine if there are gait-related variables that predispose an individual to falling after a slip. Thirty-three young and barefoot adults, protected by an instrumented safety harness, were subjected to a single slipping trial following a series of unperturbed walking trials. The slip was induced when the bare foot contacted a vinyl sheet coated with mineral oil. Lower extremity kinematics were acquired using a video-based motion capture system. Fourteen and 12 subjects could be unambiguously categorized as having fallen or recovered, respectively. Four variables demonstrated significant between-group differences and two were used to compute the probability of the slip outcome using logistic regression. The variables were the displacement of the foot during the slip and the angle of the shank relative to the ground at the instant of ground contact just prior to the slip. Separate univariate logistic regressions using each variable were significant and correctly classified about 70% of the slip outcomes. The results demonstrated that previously published values for the displacement and velocity of the slipping foot, 10cm and 50cm/s, respectively, may not accurately represent the upper limits beyond which recovery is not possible. The results also demonstrated that heel-strike angle, reflective of stride length, exerts a significant influence on the outcome of a slip. PMID- 10831755 TI - Sensitivity of periprosthetic stress-shielding to load and the bone density modulus relationship in subject-specific finite element models. AB - Subject-specific finite element (FE) computer models of the proximal femur in hip replacement could potentially predict stress-shielding and subsequent bone loss in individual patients. Before such predictions can be made, it is important first to determine if between subject differences in stress-shielding are sensitive to poorly defined parameters such as the load and the bone material properties. In this study we investigate if subject-specific FE models provide consistent stress-shielding patterns in the bone, independent of the choice of the loading conditions and the bone density-modulus relationship used in the computer model. FE models of two right canine femurs with and without implants were constructed based on contiguous computed tomography (CT) scans so that subject-specific estimates of stress-shielding could be calculated. Four different loading conditions and two bone density-modulus relationships were tested. Stress-shielding was defined as the decrease of strain energy per gram bone mass in the femur with the implant in place relative to the intact femur. The analyses showed that for the four loading conditions and two bone density modulus relationships the difference in stress-shielding between the two subjects was essentially constant (1% variation) when the same loading condition and density-modulus relationship was used for both subjects. The severity of stress shielding within a subject was sensitive to these input parameters, varying up to 20% in specific regions with a change in loading conditions and up to 10% for a change in the assumed density-modulus relationship. We conclude that although the choice of input parameters can substantially affect stress-shielding in an individual, this choice had virtually no effect on the relative differences in femoral periprosthetic stress-shielding between individuals. Thus, while care should be taken in the interpretation of the absolute value of stress-shielding calculated with these type of models, subject-specific FE models may be useful for explaining the variation in bone adaptation responsiveness between different subjects in experimental or clinical studies. PMID- 10831756 TI - Patellar cartilage deformation in vivo after static versus dynamic loading. AB - The objective of this study was to test the hypothesis that static loading (squatting at a 90 degrees angle) and dynamic loading (30 deep knee bends) cause different extents and patterns of patellar cartilage deformation in vivo. The two activities were selected because they imply different types of joint loading and reflect a realistic and appropriate range of strenuous activity. Twelve healthy volunteers were examined and the volume and thickness of the patellar cartilage determined before and from 90 to 320s after loading, using a water excitation gradient echo MR sequence and a three-dimensional (3D) distance transformation algorithm. Following knee bends, we observed a residual reduction of the patellar cartilage volume (-5.9+/-2.1%; p<0.01) and of the maximal cartilage thickness ( 2.8+/-2.6%), the maximal deformation occurring in the superior lateral and the medial patellar facet. Following squatting, the change of patellar cartilage volume was -4.7+/-1.6% (p<0.01) and that of the maximal cartilage thickness 4.9+/-1.4% (p<0.01), the maximal deformation being recorded in the central aspect of the lateral patellar facet. The volume changes were significantly lower after squatting than after knee bends (p<0.05), but the maximal thickness changes higher (p<0.05). The results obtained in this study can serve to validate computer models of joint load transfer, to guide experiments on the mechanical regulation of chondrocyte biosynthesis, and to estimate the magnitude of deformation to be encountered by tissue-engineered cartilage within its target environment. PMID- 10831757 TI - Closed-loop control of muscle length through motor unit recruitment in load moving conditions. AB - Neuroprostheses aimed at restoring lost movement in the limbs of spinal cord injured individuals are being developed in this laboratory. As part of this program, we have designed a digital proportional-integral-derivative controller integrated with a stimulation system which effects recruitment of motor units according to the size principle. This system is intended to control muscle length while shortening against fixed loads. Feline sciatic nerves were exposed and stimulated with ramp, triangular, sinusoidal, staircase and random signals as test inputs. Changes in muscle length and effective time delay under different conditions were measured and analyzed. Differences of tracking quality between open- and closed-loop conditions were examined through analysis of variance as well as the differences between small (250g) and large (1kg) loads. The results showed that parameters used to compare muscle length output to the input signals were dramatically improved in the closed-loop trials as compared to the open-loop condition. Mean squared correlation coefficients between input and output signals for ramp signals increased by 0.019, and for triangular signals by 0.12. Mean peak cross correlation between input and output signals for sinusoidal waveforms increased by 0.06, with decreases in time to peak cross correlation (effective time delay) from 195 to 38ms. In slow random signals (power up to 0.5Hz), peak cross correlation went from 0.74 to 0.89, and time-to-peak cross correlation decreased from 205 to 55ms. In fast random signals (power up to 1Hz), peak cross correlation went from 0.82 to 0.89, and time-to-peak cross correlation from 200 to 65ms. For staircase signals, both rise times and mean steady-state errors decreased. It was found that, once the length range was set, the load weight had no effect on tracking performance. Analysis of mean square error demonstrated that for all signals tested, the feedback decreased the tracking error significantly, whereas, again, load had no effect. The results suggest that tracking is vastly improved by using a closed-loop system to control muscle length, and that load does not affect the quality of signal tracking as measured by standard control system analysis methods. PMID- 10831758 TI - Determination of axial rotation angles of limb segments - a new method. AB - This paper describes a new method of determining the axial rotation angle of a limb segment during three-dimensional movement. Instead of describing the three dimensional rotation by a three-step rotation (Euler/Cardan angles), a one-step rotation (instantaneous screw axis), or a non-step rotation (floating axis method), the new method uses a two-step rotation to describe the three dimensional rotation of the limb segment: the rotation of the long axis of the limb segment about a specific axis passing through the proximal joint centre and perpendicular to the long axis of the limb segment, and the axial rotation about the long axis. A short review of previous methods followed by a full description of the principle of the new method with detailed derivation of some important equations (Appendices), comparison with Euler/Cardan angles and a simple experimental demonstration are given in this paper. A method of solving the gimbal-lock problem when using this method is also proposed. PMID- 10831759 TI - The mechanics of cell sorting and envelopment. AB - Aggregates of embryonic cells undergo a variety of intriguing processes including sorting by histological type and envelopment of cell masses of one type by another. It has long been held that these processes were driven by differential adhesions, as embodied in the famous differential adhesion hypothesis (DAH). Here, we use analytical mechanics to investigate the forces that are generated by various sub-cellular structures including microfilaments, cell membranes and their associated proteins, and by sources of cell-cell adhesions. We consider how these forces cause the triple junctions between cells to move, and how these motions ultimately give rise to phenomena such as cell sorting and tissue envelopment. The analyses show that, contrary to the widely accepted DAH, differential adhesions alone are unable to drive sorting and envelopment. They show, instead, that these phenomena are driven by the combined effect of several force generators, as embodied in an equivalent surface or interfacial tension. These unconventional findings follow directly from the relevant surface physics and mechanics, and are consistent with well-known cell sorting and envelopment experiments, and with recent computer simulations. PMID- 10831760 TI - Resonant frequencies of arms and legs identify different walking patterns. AB - The present study is aimed at investigating changes in the coordination of arm and leg movements in young healthy subjects. It was hypothesized that with changes in walking velocity there is a change in frequency and phase coupling between the arms and the legs. In addition, it was hypothesized that the preferred frequencies of the different coordination patterns can be predicted on the basis of the resonant frequencies of arms and legs with a simple pendulum model. The kinematics of arms and legs during treadmill walking in seven healthy subjects were recorded with accelerometers in the sagittal plane at a wide range of different velocities (i.e., 0.3-1. 3m/s). Power spectral analyses revealed a statistically significant change in the frequency relation between arms and legs, i.e., within the velocity range 0.3-0.7m/s arm movement frequencies were dominantly synchronized with the step frequency, whereas from 0.8m/s onwards arm frequencies were locked onto stride frequency. Significant effects of walking speed on mean relative phase between leg and arm movements were found. All limb pairs showed a significantly more stable coordination pattern from 0.8 to 1.0m/s onwards. Results from the pendulum modelling demonstrated that for most subjects at low-velocity preferred movement frequencies of the arms are predicted by the resonant frequencies of individual arms (about 0.98Hz), whereas at higher velocities these are predicted on the basis of the resonant frequencies of the individual legs (about 0.85Hz). The results support the above-mentioned hypotheses, and suggest that different patterns of coordination, as shown by changes in frequency coupling and phase relations, can exist within the human walking mode. PMID- 10831761 TI - Impact severity in self-initiated sits and falls associates with center-of gravity excursion during descent. AB - Although the energy available during a fall from standing greatly exceeds that required to produce hip fracture, this occurs in only about 2% of falls in the elderly. This is thought to be due in part to one's ability to reduce the vertical impact velocity (nu(nu)) and kinetic energy (KE(nu)) of the body through energy absorption in the lower extremity muscles during descent. The present study tested the hypothesis that the magnitude and percent attenuation in nu(nu) and KE(nu) associate with the horizontal and vertical excursion of the body's center-of-gravity during descent. Measures were acquired of whole-body kinematics and lower extremity kinetics as young subjects underwent backward descents involving vertical drops of either thigh length (SIT) or lower extremity length (FALL), and horizontal pelvis excursions of either 33 or 66% of lower extremity length. In all trials, subjects attempted to "land as softly as possible." While attenuation in nu(nu) and KE(nu) (which averaged 62 and 92% respectively), did not associate with trial type, raw magnitudes of these parameters did, with nu(nu) averaging 2-fold greater, and KE(nu) averaging 6-fold greater, in 66% FALL than in 33% SIT or 66% SIT trials. This was due to a rapid increase in downward velocity accompanying the final stage of descent in 66% SIT and 66% FALL trials, which coincided with the knee moving posterior to the ankle. Accordingly, severe impacts likely accompany not only large fall heights, but also falls where the feet are thrown rapidly forward, as during a backward slip. PMID- 10831762 TI - The effect of hamstring muscle compensation for anterior laxity in the ACL deficient knee during gait. AB - The hamstring muscles have been recognized as an important element in compensating for the loss of stability in the ACL-deficient knee, but it is still not clear whether the hamstring muscle force can completely compensate for the loss of ACL, and the consequences of increased hamstring muscle force. A two dimensional anatomical knee model in the sagittal plane was developed to examine the effect of various levels of hamstring muscle activation on restraining anterior tibial translation in the ACL-deficient knee during level walking. The model included the tibiofemoral and patellofemoral joints, four major ligaments, the medial capsule, and five muscle units surrounding the knee. Simulations were conducted to determine anterior tibial translation and internal joint loading at a single selected position when the knee was under a peak external flexion moment during early stance phase of gait. Incremental hamstring muscle forces were applied to the modeled normal and the ACL-deficient knees. Results of simulations showed that the ACL injury increased the anterior tibial translation by 11.8mm, while 56% of the maximal hamstring muscle force could reduce the anterior translation of the tibia to a normal level during the stance phase of gait. The consequences of increased hamstring muscle force included increased quadriceps muscle force and joint contact force. PMID- 10831763 TI - Impact of the porous microstructure on the overall elastic properties of the osteonal cortical bone. AB - Mechanical properties of bones are largely determined by their microstructure. The latter comprises a large number of diverse pores. The present paper analyzes a connection between structure of the porous space of the osteonal cortical bone and bone's overall anisotropic elastic moduli. The analysis is based on recent developments in the theory of porous materials that predict the anisotropic effective moduli of porous solids in terms of pores' shapes, orientations and densities. Bone's microstructure is modeled using available micrographs. The calculated anisotropic elastic constants for porous cortical bone are, mostly, in agreement with available experimental data. The influence of each of the pore types on the overall moduli is examined. The results of the analysis can also be used to estimate the extent of mineralization (hydroxyapatite content) if the overall porosity and the effective moduli are known and, vice versa, to estimate porosity from the measured moduli and the extent of mineralization. PMID- 10831764 TI - Factors influencing the output of an implantable force transducer. AB - The objective of this study was to evaluate the performance of the Arthroscopically Implantable Force Probe (AIFP; MicroStrain, Burlington VT) for measuring force in a patellar tendon graft. Transducer drift, reproducibility of output due to the number of loading cycles and device location, and sensitivity to the tendon cross-sectional area were investigated. The AIFP was initialized, and then implanted into five human patellar tendon grafts three times; twice within the same location and once in a different location. The tendons were cyclically loaded in uniaxial tension for 500 cycles in each insertion site. The AIFP was then removed from the tendon and the baseline output was remeasured. It was determined that transducer drift was negligible. The relationship between the tensile load applied to the graft and AIFP output was quadratic and specimen dependent. The cyclic load response of the tendon-AIFP interface demonstrated a 24.9% decrease over the first 20 loading cycles, and subsequent cycling yielded relatively reproducible output. The output of the transducer varied when it was removed from the tendon and then reimplanted in the same location (range 3.7-109. 4% error), as well as in the second location (range 1.5-202.8% error). No correlation was observed between the cross-sectional area of the tendon and transducer output. This study concludes that implantable force probes should be used with caution and calibrated without removing the transducer from the graft. PMID- 10831765 TI - Finite element estimates of interface stress in the trans-tibial prosthesis using gap elements are different from those using automated contact. AB - When compared with automated contact methods of finite element (FE) analyses, gap elements have certain inherent disadvantages in simulating large slip of compliant materials on stiff surfaces. However, automated contact has found limited use in the biomechanical literature. A non-linear, three-dimensional, geometrically accurate, FE analysis of the trans-tibial limb-socket prosthetic system was used to compare an automated contact interface model with a gap element model, and to evaluate the sensitivity of automated contact to interfacial coefficient of friction (COF). Peak normal stresses and resultant shear stresses were higher in the gap element model than in the automated contact model, while the maximum axial slip was less. Under proximally directed load, compared with automated contact, gap elements predicted larger areas of stress concentration that were located more distally. Gap elements did not predict any relative slip at the distal end, and also transmitted a larger proportion of axial load as shear stress. Both models demonstrated non -linear sensitivity to COF, with larger variation at lower magnitudes of COF. By imposing physical connections between interface surfaces, gap elements distort the interface stress distributions under large slip. Automated contact methods offer an attractive alternative in applications such as prosthetic FE modeling, where the initial position of the limb in the socket is not known, where local geometric features have high design significance, and where large slip occurs under load. PMID- 10831766 TI - The use of basis functions in modelling joint articular surfaces: application to the knee joint. AB - This article introduces a new method to represent bone surface geometry for simulations of joint contact. The method uses the inner product of two basis functions to provide a mathematical representation of the joint surfaces. This method guarantees a continuous transition in the direction of the surface normals, an important property for computation of joint contact. Our formulation handles experimental data that are not evenly distributed, a common characteristic of digitized data of musculoskeletal morphologies. The method makes it possible to represent highly curved surfaces, which are encountered in many anatomical structures. The accuracy of this method is demonstrated by modeling the human knee joint. The mean relative percentage error in the representation of the patellar track surface was 0.25% (range 0-1.56%) which corresponded to an absolute error of 0.17mm (range 0-0.16mm). PMID- 10831767 TI - Selective regional loss of exocytotic presynaptic vesicle proteins in Alzheimer's disease brains. AB - We tested whether regional or selective alterations in presynaptic proteins occur in Alzheimer's disease (AD) and correlate with tests of cognitive function. We measured the levels of seven presynaptic proteins (synaptobrevin, synaptotagmin, SNAP-25, syntaxin, SV2, Rab3a, and synapsin I) by immunoblotting in postmortem tissue from four brain regions (hippocampus, entorhinal cortex, caudate nucleus, and occipital cortex). Three subject groups were studied: AD, possible/early AD (p-AD), and age-matched controls. Synaptobrevin and synaptotagmin were significantly reduced (29%, P<0.08; 38%, P<0. 07) in hippocampus in p-AD compared to controls. In definite AD compared to controls, selective regional reductions in vesicle proteins were found: synaptobrevin (46%, P<0.05), synaptotagmin (52%, P<0.01), and Rab3a (30%, P<0.05) in hippocampus; synaptobrevin (31%, P<0.01), synaptotagmin (15%, P<0.05), and Rab3a (44%, P<0.05) in entorhinal cortex. In contrast, the levels of two vesicle proteins (synapsin I and SV2) and two presynaptic membrane proteins (syntaxin and SNAP-25) were similar to controls. Synaptobrevin was the only vesicle protein reduced in AD in all four brain regions (occipital cortex 37%, P<0.05; caudate nucleus 31%, P<0.05). By univariate analysis of all cases, Mini-Mental State Examination, Blessed (BIMC) and Free Recall scores were strongly correlated with reduced levels of synaptic vesicle proteins synaptobrevin, synaptotagmin, and Rab3a in hippocampus and entorhinal cortex. These results suggest that there are selective and early defects in presynaptic vesicle proteins, but not synaptic plasma membrane proteins in AD and that defects correlate with cognitive dysfunction in this disease. PMID- 10831768 TI - Increased lipid peroxidation in the brains of aceruloplasminemia patients. AB - Aceruloplasminemia is characterized by excessive neurovisceral accumulation of iron due to mutations in the ceruloplasmin gene. We report that the levels of malonaldehyde (MDA) and 4-hydroxynonenals (4-HNE) were elevated both in the frontal cortex and putamen in two patients with aceruloplasminemia compared with those in controls. Immunohistochemical analysis with an antibody raised against HNE-modified protein showed a large number of immunoreactive neurons and glias in the brain of a patient. MDA and 4-HNE are good indicators of lipid peroxidation because they are generated in the process of oxidation of polyunsaturated fatty acids. The data strongly suggest that the increase of oxygen free radicals associated with excessive iron accumulation plays an important pathogenic role in the process of neuronal cell death in the brains of patients with aceruloplasminemia. PMID- 10831769 TI - Antibodies to GM1(NeuGc) in Guillain-Barre syndrome after ganglioside therapy. AB - N-Glycolylneuraminic acid-containing GM1 [GM1(Gc)] is a molecule for serum antibodies in patients with Guillain-Barre syndrome (GBS). To clarify the pathogenesis of GBS after treatment with bovine brain ganglioside, we investigated the presence of anti-GM1(Gc) antibody in patients who developed GBS after ganglioside injection. Serum samples were taken from nine Italian patients with GBS after ganglioside therapy as well as from untreated Italian (n=30) and Japanese (n=131) GBS patients. Bovine brain gangliosides fractionated in a column were used as antigens, and binding of serum IgG or IgM was examined. An absorption study of IgG anti-GM1(Gc) antibody was made with GM1, asialo-GM1, GM2, GD1a, and GD1b. Four of the nine patients who developed GBS after being administered gangliosides had IgG anti-GM1(Gc) antibodies. Anti-GM1(Gc) IgG antibody frequencies were higher in patients with GBS after ganglioside therapy than in those who were untreated. Rates of absorption of IgG anti-GM1(Gc) antibodies by GM1 were significantly higher (except for asialo-GM1 and GD1b) than by GM2 and GD1a. The presence of GM1(Gc) was confirmed in bovine brain immunochemically using cholera toxin and Hanganutziu-Deicher antibody. Secondary ion mass spectra showed that the structure of the ganglioside was consistent with that of GM1(Gc). GM1(Gc) was recognized more frequently in sera from patients who developed GBS after ganglioside therapy than in sera from untreated GBS patients. Because N-glycolylneuraminic acid-containing gangliosides seem to be highly immunogenic in humans, GM1(Gc) may act as an immunogen in some patients who develop GBS following ganglioside therapy. PMID- 10831770 TI - Oxidative injury is present in Purkinje cells in patients with olivopontocerebellar atrophy. AB - To verify the presence of lipid peroxidation products in spinocerebellar degeneration (SCD), the cerebella from eight patients with olivopontocerebellar atrophy (OPCA) and six non-OPCA patients were immunohistochemically investigated with 4-hydroxy-2-nonenal (HNE) antibody. On average, 84.6% of Purkinje cells were positively or strongly positively immunostained in OPCA patients while only 15.5% were positive in non-OPCA patients. Other cells in the molecular and granular layers showed no obvious immunoreactivity. These data suggest that a lipid peroxidation product is present in Purkinje cells of OPCA patients and that oxidative stress may play an important role in the degeneration process of SCD. PMID- 10831771 TI - IgG allotypes and subclasses in Norwegian patients with multiple sclerosis. AB - Multiple sclerosis (MS) is a multifactorial disease in which genetic and environmental factors apparently have a major influence on the susceptibility and course of the disease. In the present study we have investigated the genetic basis and subclass levels of IgG in MS. Hundred and thirty-six Norwegian patients with MS and 92 controls were genotyped for IgG allotypes of the GM and KM systems. IgG and IgG subclasses were quantified in sera from 115 MS patients and 20 controls. Neither GM nor KM allo-, haplo- or genotypes were significantly correlated with susceptibility, severity or course of the disease. The G1M (3) (3), G2M (23) (23) and G3M (5) (5) allotypes were significantly correlated with high serum levels of IgG3, whereas high IgG2 levels were correlated with G1M (3) (3) and G2M (23) (23) in both patients and controls. Serum levels of IgG subclasses were not significantly correlated with course or severity of the disease. The results indicate no major role for IgG allotypes or IgG subclass levels in the pathogenesis of MS. PMID- 10831772 TI - The Scandinavian Multi-Infarct Dementia Trial: a double-blind, placebo-controlled trial on nimodipine in multi-infarct dementia. AB - Vascular dementia is a major cause of mental and physical disability in Western countries. Treatment of vascular dementia is currently based on the recognition and control of vascular risk factors, while specific drugs have not been approved yet. The aim of the present multinational, double-blind, placebo-controlled study was to evaluate the safety and efficacy of nimodipine administered for as long as 26 weeks in improving cognition or slowing cognitive deterioration in patients defined as having multi-infarct dementia (DSM-III-R criteria). Two hundred and fifty-nine patients were included (128 nimodipine, 131 placebo), and 251 were available for the intention-to-treat analysis. No significant difference between drug-treated and placebo patients was noted on the Gottfries-Brane-Steen scale score (primary efficacy criterion), the remaining neuropsychological tests (Zahlen-Verbindungs-Test, Fuld-Object-Memory Evaluation, Word Fluency Test, Digit Span, Mini-Mental State Examination), and the functional scales (index of Activity of Daily Living, Instrumental Activity of Daily Living, Rapid Disability Scale, Clinical Dementia Rating), although the majority of changes were in favor of the active drug group. A lower incidence of cerebrovascular and cardiac events was observed in the nimodipine-treated patients in comparison with the placebo group. This study failed to show a significant effect of nimodipine on cognitive, social or global assessments in patients defined as affected by multi-infarct dementia according to the DSM-III-R criteria. A post-hoc analysis (presented in an accompanying paper) suggests that nimodipine may have a favorable effect in the subgroup of patients defined as affected by subcortical (small vessel) vascular dementia. PMID- 10831773 TI - Efficacy and safety of nimodipine in subcortical vascular dementia: a subgroup analysis of the Scandinavian Multi-Infarct Dementia Trial. AB - In Western countries, vascular dementia (VaD) is the most common form of cognitive deterioration after Alzheimer's disease. Therapeutic trials in VaD have so far failed to yield satisfactory results. One explanation of this failure may be the etiological and clinical heterogeneity of the included patients. Patients with subcortical VaD, defined on a clinical and radiological basis, may constitute a more homogeneous group. Thus, we conducted a post-hoc subgroup analysis of the Scandinavian Multi-Infarct Dementia Trial that evaluated the efficacy and safety of oral nimodipine administered for 6 months in 259 patients. The original patients sample was divided on the basis of head CT in those with subcortical VaD (n=92, 45 nimodipine, 47 placebo) and those with multi-infarct dementia (n=167, 83 nimodipine, 84 placebo). While in the total trial population a treatment effect could not be proved, in this subgroup analysis, the subcortical VaD patients treated with nimodipine performed better on the majority of neuropsychological tests and functional scales in comparison with patients on placebo. No trend could be evidenced in the multi-infarct dementia patients. Treatment efficacy was in particular suggested for the Zahlen-Verbindungs-Test, Fuld-Object-Memory Evaluation, Word Fluency, and for the Instrumental Activities of Daily Living scale. The results did not reach statistical significance in this small sample. Our study preliminarily indicates that nimodipine could be effective in patients with small vessel subcortical VaD and supports the rationale for a further controlled and adequately powered trial to test nimodipine in patients with subcortical VaD. PMID- 10831774 TI - Influence of immobilization upon calcium metabolism in the week following hemiplegic stroke. AB - Hip fractures on the paretic side are a serious post-stroke complication and may result from disuse hemiosteopenia, hypovitaminosis D, and an increasing risk of falls. To evaluate short-term immobilization effects, we assessed calcium metabolism in 89 patients 1 week after the hemiplegic stroke and in 36 controls. Patient activity was rated using the Barthel index (BI). Sera from stroke patients and control subjects were assayed for ionized calcium, parathyroid hormone (PTH), 25-hydroxyvitamin D (25-OHD), 1, 25-dihydroxyvitamin D (1,25 (OH)(2)D), bone Gla protein (BGP; a bone formation marker) and pyridinoline cross linked carboxy-terminal telopeptide of type I collagen (ICTP; a bone resorption marker). Patients' serum concentrations of ionized calcium and ICTP were higher than in controls and correlated negatively with BI; their BGP concentrations were low, correlating positively with BI. Concentrations of serum 25-OHD, 1,25 (OH)(2)D, and PTH also were low; serum 25-OHD was at a deficient level (<10 ng/ml) in nine patients (10%), an insufficient level (10-20 ng/ml) in 56 (63%), and a sufficient level (>20 ng/ml) in only 24 (27%). PTH correlated negatively with calcium and 1,25-(OH)(2)D. Hypovitaminosis D is common in acute stroke patients. Immobilization from acute hemiplegia can increase bone resorption and serum calcium, and inhibit PTH secretion and 1,25-(OH)(2)D production to add to the effects of hypovitaminosis D. PMID- 10831775 TI - A severe case of subacute sarcoid myositis. AB - A 46-year-old woman presented progressive proximal weakness and dysphagia. Her serum creatine kinase and myoglobin levels were markedly elevated. Chest X-rays revealed bilateral swelling of the hilar lymph nodes. Needle electromyography demonstrated active denervation and early recruitment. MRI of her skeletal muscle showed focal high intensities on T1-weighted images that were associated with diffusely increased signal intensities on T2-weighted images. Muscle biopsy revealed infiltration of inflammatory cells associated with non-caseating granulomas, and there was widespread segmental fiber necrosis, where necrotic fibers appeared regardless of these granulomas. Immunohistochemical analysis of the surface markers of the infiltrating cells showed CD68- and CD4-positive cells infiltrating into the central area of the granuloma, while CD8-positive cells infiltrating into the endomysium and the periphery of the granulomas. The characteristic histology of the granuloma confirmed the diagnosis of sarcoidosis. The diffuse muscle pathology was consistent with the patient's severe clinical course. PMID- 10831776 TI - Hyper-reflexia without spasticity after unilateral infarct of the medullary pyramid. AB - Whether or not a lesion confined to the pyramidal tract produces spasticity in humans remains an unresolved controversy. We have studied a patient with an ischemic lesion of the right medullary pyramid, using objective measures of hyper reflexia, spasticity, and weakness. Electromyographic activity (EMG) of the biceps muscles was recorded under the following conditions: (1) in response to a tendon tap with an instrumental reflex hammer, (2) in response to imposed quick stretch with motion analysis, and (3) during an isometric holding task. Hyper reflexia of the involved arm in response to tendon tap was shown to be due primarily to an increase in the gain of the reflex arc. No velocity-dependent increase in the response to quick stretch of the involved arm was present. This was consistent with the absence of detectable spasticity on the clinical exam. These findings suggest that a lesion confined to the medullary pyramid can give rise to weakness and hyper-reflexia without causing spasticity. Moreover, these findings suggest that different anatomical substrates may underlie the clinical phenomena of hyper-reflexia and spasticity. PMID- 10831777 TI - Influence of relative layer height and testing method on the failure mode and origin in a bilayered dental ceramic composite. AB - OBJECTIVES: The purpose of this study was to examine the influence of testing method (uniaxial and biaxial) and relative layer heights on the failure origin and failure mode of bilayered ceramic composite beams and disks composed of In Ceram and Vitadur Alpha porcelain. METHODS: Beams and disks were fabricated, with relative layer heights of 1:2, 1:1, and 2:1, respectively, for In-Ceram and Vitadur Alpha porcelain. Ninety specimens each (thirty 1:2, thirty 1:1, and thirty 2:1) were tested in 3-point, 4-point-1/4-point, and biaxial ring-on-ring testing apparatuses. Fractography was used to categorize failure origins as either surface or interfacial, and failure modes as delamination or nondelamination. RESULTS: Surface and interfacial failure origins were observed in 3-point and biaxial disk test specimens, but not 4-point-1/4-point specimens where only surface failures occurred. None of the "clinically similar" specimens (1:2) failed at the interface. All testing methods resulted in delamination of Vitadur Alpha from the In-Ceram, while only 3-point and biaxial disk testing methods resulted in crack propagation through the composite interface without delamination. SIGNIFICANCE: Varying relative layer heights or varying testing method in laminate composite tensile specimens can affect failure mode and failure origin. PMID- 10831778 TI - The effect of post-cure heating on residual, unreacted monomer in a commercial resin composite. AB - OBJECTIVES: This paper examined the influence of post-cure temperature on the amount of unreacted monomer remaining in a commercial light-cured resin composite restoration following initial light-curing and subsequent post-cure heating. METHODS: Discs of composite were light-cured and then subjected to immediate post cure heating (50, 75, 100, or 125 degrees C for 7 min) or were left unheated (control). They were then placed in a solvent for two weeks at 37 degrees C to extract the unreacted monomer. HPLC analysis was used to determine the amounts of TEGDMA, BIS-GMA, and ethoxylated BIS-GMA remaining after the different treatments. The amounts of each monomer leaching were compared using ANOVA with respect to the different curing treatments. RESULTS: Even the lowest post-cure heat treatment (50 degrees C) resulted in 80% reduction in remaining, unreacted TEGDMA, 75% reduction in BIS-GMA, and 77% lower ethoxylated BIS-GMA than the light-cured only control. Post-cure heating at 75 degrees C and above resulted in the lowest amount of each type monomer remaining uncured in the polymer and did not significantly decrease with an increase in post-cure temperature for the most part. SIGNIFICANCE: One of the main benefits of post-cure heating of resin composite restorations could be the enhancement of biocompatibility of these restorations as a result of the significant decrease in potentially leachable, unreacted monomer. PMID- 10831779 TI - Indentation modulus and hardness of whisker-reinforced heat-cured dental resin composites. AB - OBJECTIVES: Recent studies showed that ceramic whisker reinforcement imparted a two-fold increase in the strength of dental composites. The aim of this study was to investigate the indentation response and measure the elastic modulus, hardness, and brittleness of whisker-reinforced heat-cured resin composites as a function of filler level, heat-cure temperature, and heat-cure duration. METHODS: Silica particles were fused onto silicon nitride whiskers to facilitate silanization and to roughen the whiskers for improved retention in matrix. Whisker filler mass fractions of 0, 20, 40, 60, 70, 74 and 79% were tested. Heat cure temperature ranged from 100 to 180 degrees C, and duration from 10 min to 24 h. A nano-indentation system enabled the measurement of elastic modulus. Fracture toughness was measured and composite brittleness index was calculated. An inlay/onlay composite and a prosthetic composite were tested as controls. RESULTS: Whisker filler level and heat-cure duration had significant effects on composite properties, while heat-cure temperature had non-significant effects. The whisker composite with 79% filler level had a modulus in GPa (mean (SD); n = 6) of 26.9 (1.0), significantly higher than 15.1 (0.2) of an inlay/onlay control, and 16.1 (0.3) of a prosthetic control (Tukey's multiple comparison test; family confidence coefficient = 0.95). The fracture toughness in MPa.m1/2 was 2.22 (0.26) for the whisker composite, higher than 0.95 (0.11) for inlay/onlay control, and (1.13 +/- 0.19) for prosthetic control. The brittleness index was (0.49 +/- 0.07) for whisker composite, lower than (1.02 +/- 0.12) for inlay/onlay control and (0.63 +/- 0.13) for prosthetic control. SIGNIFICANCE: Whisker filler level had a profound influence, heat-cure duration had significant effects, while temperature did not have significant effects, on the properties of whisker composite. The whisker composite had significantly higher elastic modulus and fracture toughness, and lower brittleness than the inlay/onlay and prosthetic controls. PMID- 10831780 TI - The influence of the modification of etched bovine dentin on bond strengths. AB - OBJECTIVE: The aim of this study was to modify demineralized bovine dentin surfaces by application of either 12.5% sodium hypochlorite (NaOCl), or 0.1% (w/w) Type I collagenase, after conditioning with phosphoric acid, to observe the demineralized surface and to investigate the effect on tensile bond strength. METHODS: The NaOCl was applied to etched dentin for 30 s, 1 or 2 min and the collagenase for 1, 3 or 6 h. A control group was used without NaOCl or collagenase treatment. Prior to bonding, treated surfaces were examined using an Atomic Force Microscope (AFM). A 2.3 mm diameter area of dentin was conditioned, treated and bonded with either One Coat Bond or Single Bond following each manufacturer's instructions, and a resin composite rod attached. Bonds were stressed in tension at a rate of 1 mm/min until failure. Mean bond strengths were calculated (MPa) and mode of failure was determined by observation at 20x magnification. Results were analyzed using multiple regression analysis and LSD test at the 95% level of confidence (n = 12). RESULTS: AFM results showed progressive changes of the surface collagen as the treatment time of NaOCl or collagenase increased. For both bonding systems, the bond strengths of 1 min NaOCl and 3 h collagenase treatments were significantly higher than the control or other treatment groups (p < 0.05). Bond failure consisted of mostly adhesive failure between dentin and resin combined with small regions exhibiting cohesive failure of resin. SIGNIFICANCE: Bond strengths were not dependent on the thickness of the hybrid layer, but rather quality of the hybrid layer. PMID- 10831781 TI - Electrochemical characteristics of high-Pd alloys in relation to Pd-allergy. AB - OBJECTIVE: The aim of this study was to determine whether the Pd-Cu-based dental ceramic alloys possess any electrochemical characteristics distinguishable from that of other Pd-containing alloys. Of all Pd-containing alloys, this particular alloy group has been linked to frequent incidence of allergy and hypersensitivity reactions. Electrochemical corrosion may instigate these reactions. METHODS: Four groups of alloys, Pd-Cu, Pd-Ga-(with and without Ag), Pd-Ag, and Au-Pd, were evaluated by traditional corrosion measurement techniques in a phosphated buffer saline solution at 20 degrees C. The electrochemical characteristics measured were: (1) 20 h open circuit potential (OCP); (2) 20 h corrosion rate (Icorr); and (3) anodic polarization (E-i) curves. RESULTS: The OCP values (232 +/- 25 mV) of the Ag-free Pd-Ga and Pd-Cu-based alloys were higher than and distinctly different from that (144 +/- 52 mV) of the Ag-containing alloys. The Icorr values of different alloys, despite varied compositions, were indistinguishable from one another. The E-i curves of all alloys were essentially similar, with the Ag containing (> 5 wt%) alloys showing a subtle difference in their anodic slope within 100 mV above their corrosion potentials. SIGNIFICANCE: The OCP values of Pd-Cu alloys and the Ag-free Pd-Ga alloy are comparable to that reported for pure Pd (239 +/- 21 mV), which indicates that during corrosion these alloys undergo dealloying and consequent Pd-enrichment on their surface. Such a condition is conducive to the release of allergenic Pd++ ions and offers a plausible explanation for the frequent incidence of hypersensitivity reactions associated with the Pd-Cu alloys. The OCP values in other alloys are attributed to dealloying followed by surface enrichment with Ag and/or Au and the possible formation of an insoluble AgCl surface film on the respective alloy surfaces. These events have the potential to suppress or prevent Pd++ ion release. Alloys showing these characteristics have seldom been linked to allergic reactions. PMID- 10831782 TI - Effect of etchant variation on wet and dry dentin bonding primed with 4 META/acetone. AB - OBJECTIVE: To collect data that explains the advantage, if any, of wet bonding versus dry bonding to dentin, and to more clearly understand the mechanism of wet bonding. METHODS: Bovine dentin samples were prepared with #600-grit paper and were divided into four groups of six each. The first six specimens were etched with 10% citric acid and 3% ferric chloride for 10 s then rinsed and blot-dried (Gr. 1: 10-3:W). The second six were etched with 10% citric acid and 3% ferric chloride then rinsed and air-dried (Gr. 2: 10-3:D). The third six were etched with 10% citric acid for 10 s, rinsed and blot-dried (Gr. 3: 10-0:W). The fourth group of six samples was etched, rinsed and air-dried (Gr. 4: 10-0:D). All groups were primed with 5% 4-methacryloyloxyethyl trimellitate anhydride (4-META) in acetone for 60 s and an acrylic rod was bonded to the samples using a 4 META/methyl methacrylate (MMA)-tri-n-butyl borane (TBB) resin. The samples were fashioned into dumbbell-shaped specimens and stressed in tension until bond failure, to determine tensile bond strengths. Fractured surfaces were examined by scanning electron microscopy. RESULTS: The mean tensile bond strengths of specimen groups were found to be 18.9 (8.1) MPa (Gr. 1: 10-3:W), 18.1 (1.7) MPa (Gr. 2: 10-3:D), 11.9 (4.4) MPa (Gr. 3: 10-0:W) and 5.4 (2.8) MPa (Gr. 4: 10 0:D). There was no statistically significant difference between Grs. 1 and 2 (p > 0.01), or between Grs. 1 and 3 (p > 0.05). The TBS of Gr. 4 (10-0:D) was significantly lower than the others (p < 0.01). The 5% 4-META in acetone primer was apparently quite effective in improving monomer impregnation into demineralized dentin resulting in increased resin content within the hybridized dentin. SIGNIFICANCE: Effective dentin bonding depended upon the etchants employed. 10-0 etching and air-drying caused the demineralized dentin to collapse in which case wet bonding became necessary to obtain good TBS data. The specimens demineralized with 10-3 did not collapse, even when air-dried; consequently both wet and dry bonding proved effective for obtaining high tensile bond strength data. PMID- 10831783 TI - Effect of furnace type and ceramming heat treatment conditions on the biaxial flexural strength of a canasite glass-ceramic. AB - OBJECTIVE: To assess the effect of different heat treatment conditions when using two different furnace types on the biaxial flexural strength (BFS) of a fluorcanasite castable glass-ceramic. METHODS: Two furnace types, one a programmable furnace (PF), the other a dental laboratory burnout furnace (DLF), were used with various ceramming times to determine their effect on the BFS of a fluorcanasite castable glass-ceramic. The glass-ceramic material was cast to produce discs of 12 mm diameter and 2 mm thickness using the lost wax casting process (n = 80). After casting, both furnace types were used to ceram the discs. Half the discs were not de-vested from the casting ring before ceramming but cerammed in situ (DLF) and half were de-vested before ceramming (PF). All the discs were given a nucleation heat treatment at 520 degrees C for 1 h and then cerammed at 860 degrees C using four heat soak times (0.5, 1, 2 and 3 h). The DLF furnace had a rate of climb of 13 degrees C/min and the PF furnace had a rate of climb of 5 degrees C/min to 520 degrees C and 3 degrees C/min to 860 degrees C. After ceramming the discs were de-vested and the BFS determined using a Lloyd 2000R tester. RESULTS: The maximum BFS values seen for both furnace types were almost identical (280 MPa), but were achieved at different heat soak times (1 h DLF, and 2 h PF). The only significant differences in BFS values for the two furnaces were between the 0.5 and 2 h heat soak times (p < or = 0.05). Individual differences were seen between results obtained from each furnace type/heat soak times evaluated (p < or = 0.05). SIGNIFICANCE: Already available dental laboratory burnout furnaces can be used to ceram fluorcanasite glass-ceramic castings to the same BFS values as more expensive and slower specialist programmable furnaces. PMID- 10831784 TI - Clinical evaluation of three adhesive systems in class V non-carious lesions. AB - OBJECTIVES: The purpose of this study was to evaluate the clinical retention of three new adhesive systems in non-carious cervical lesions during a 3-year period. METHODS: The adhesive systems, a three-step (EBS/Pertac Hybrid), a one bottle resin bonding agent (One-Step/Pertac Hybrid) and a resin-modified glass ionomer cement (Fuji II LC) were placed in 148 non-carious cervical lesions, 87 with sclerotic dentin and 61 non-sclerotic. Of the sclerotic lesions treated with the two resin bonding systems, 37 were slightly roughened with a diamond bur before conditioning. The restorations were evaluated every 6 months during a 3 year period. RESULTS: All except six restorations were evaluated during the 3 years. The cumulative loss rates for EBS, One-Step and Fuji II LC were, at 1 year: 2, 24 and 2% and at 3 years: 10, 49, 7%, respectively. The one-bottle adhesive showed significantly more failures. The five lost EBS restorations were found in non-sclerotic lesions, while the three lost Fuji II LC restorations had been placed in sclerotic lesions. For the One-Step material the loss frequency for non-sclerotic versus sclerotic lesions was 31.8 and 65.2%, respectively. Slight roughening of the sclerotic dentin surfaces with a diamond bur did not increase retention of the restorations. SIGNIFICANCE: The three-step resin adhesive and the RMGIC showed clinically acceptable retention rates, while a high failure rate was registered for the one-bottle adhesive. PMID- 10831785 TI - Light intensity effects on resin-composite degree of conversion and shrinkage strain. AB - OBJECTIVE: The aim of this study was to investigate the relationship between the degree of conversion (DC) and shrinkage strain values of two representative resin composites when irradiated by two light intensity levels, for different time periods. METHODS: The DC of Z100 and Tetric Ceram was measured by FTIR spectroscopy immediately post-cure and shrinkage strain values were obtained continuously for 30 min from photoinitiation with the bonded-disk technique. All samples were photopolymerized at an initial temperature of 23 degrees C with the Elipar Highlight unit under the following modes of variable light intensity (I) 40 s at 750 mW/cm2; (II) a two-step 'soft-start' irradiation mode of 10 s at 200 mW/cm2 plus 30 s at 750 mW/cm2; (III) 40 s at 200 mW/cm2; and (IV) 10 s at 200 mW/cm2. Temperature effects of the light and curing exotherm were not eliminated. RESULTS: Mode II of irradiation did not reduce the DC for each material, relative to the values obtained with mode I (p > 0.05). The corresponding shrinkage strain values after 30 min were also not significantly different (p > 0.05). However, the effects of reduced light intensity levels for 10 s and 40 s (modes III and IV), were in agreement with previous findings implying reduced shrinkage strain levels. For both resin composites, data from all curing times and light intensities gave a linear regression relationship (with r2 > 0.99) between the bonded-risk shrinkage strain versus DC values. SIGNIFICANCE: For the materials and light-intensities studied, the correlation between DC and shrinkage strain values means that some reductions in the problems of shrinkage may be achieved by an acceptable reduction in DC. However, this must also be consistent with generating adequate curing performance in the resin-network. Nevertheless, DC is not the exclusive parameter describing network character and a fuller understanding of the latter is an important research goal. PMID- 10831786 TI - Changes in cell phospholipid metabolism in vitro in the presence of HEMA and its degradation products. AB - OBJECTIVES: Diacylglycerol-kinase (DAG-kinase) is an enzyme that phosphorylates diacylglycerol (DAG) to phosphatidic acid (PA), which serves as a precursor to phosphoglycerides involved in cell signaling or as cell membrane structural components. DAG-kinase can be inhibited by diacylethylene glycols (DAEG). We hypothesize that 2-hydroxyethyl methacrylate (HEMA) may alter phosphorylation of DAG to PA following intracellular formation of DAEG. METHODS: Cultured rabbit kidney (RK13) epithelial cells were treated with HEMA, EG, or known inhibitors of DAG-kinase for 24 h, then exposed to [32P]O4- in the presence of a synthetic diacylglycerol for 2 h. Other cultures were radiolabeled with [3H]-oleic acid for 24 h, then exposed to HEMA for an additional 24 h. The cells were harvested and the lipids extracted. Radioactive lipids were separated by thin layer chromatography, located by autoradiography, and quantitated as cpm/ug protein. Cell cultures treated with HEMA were homogenized and the DAG-kinase activity was assayed and expressed as cpm/ug protein. Data were analyzed by one-way ANOVA and Newman-Keuls Multiple Comparison Test. RESULTS: Cultures exposed to HEMA or known DAG-kinase inhibitors exhibited reduced incorporation of radioactivity in the PA fraction compared to control cultures. Direct assays of DAG-kinase activity from cells exposed to HEMA demonstrated decreased enzyme activity. Evaluation of cell phospholipid synthesis showed altered formation of phosphatidylethanolamine and phosphatidylcholine. SIGNIFICANCE: Results suggest that HEMA impairs formation of PA, possibly by acylation of EG released by hydrolysis of the HEMA and resultant production of the inhibitor DAEG. The decreased availability of PA may alter PA dependent cell structural lipid pathways and lipid-dependent signaling pathways, altering cell growth. PMID- 10831787 TI - Influence of time and adhesive system on the extrusion shear strength between feldspathic porcelain and bovine dentin. AB - OBJECTIVES: The behavior of dual-cure cements over time remains unclear. This study evaluated the extrusion shear strength of the bond between feldsphatic porcelain and bovine dentin at different time intervals, using three adhesive systems based on dual-cure cements and one based on a self-cure cement. METHODS: The adhesive systems evaluated included: C&B/One-Step, Enforce/Prime&Bond NT Dual Cure, RelyX ARC/Single Bond and Variolink II/Syntac SC. Discs of bovine root dentin, 2.5 mm thick, had the root canal prepared with a standardized taper. Porcelain truncated cones etched with 4% hydrofluoric acid and silanized were bonded into the perforations. The extrusion shear test was performed after 15 min, 4 h, 12 h, 24 h, and 7 days. Data were analyzed using the Weibull distribution. The failure mode was accessed by stereomicroscope and SEM. RESULTS: Statistical differences were found between 15 min and 7 days for Variolink II/Syntac SC and RelyX ARC/Single Bond, and between 15 min and 24 h for Enforce/Prime&Bond NT Dual-Cure. For C&B/One-Step, difference was found between 15 min and 4 h. Between 24 h and 7 days, only the system RelyX ARC/Single Bond showed a significant increase in the characteristic strength. SEM analysis revealed that the failure involved the hybrid layer/dentin or the hybrid layer/adhesive interfaces, regardless of the time interval and adhesive system. SIGNIFICANCE: High characteristic strengths were observed after 15 min when dual cure cements were used. In general, the values found at 24 h or 7 days were higher than at 15 min. However, there was always a considerable probability of bonding failure at low stress levels for all the systems tested. PMID- 10831788 TI - Production of reactive oxygen species by hemocytes of Biomphalaria glabrata: carbohydrate-specific stimulation. AB - Recognition of specific carbohydrate structures, which occur commonly on the surfaces of invading pathogens, is thought to elicit internal defense mechanisms in invertebrates. To investigate the nature of carbohydrates that evoke a defensive response in hemocytes of the gastropod Biomphalaria glabrata, we tested eight different carbohydrates, conjugated to bovine serum albumin (BSA), for generation of reactive oxygen species (ROS). Six of the carbohydrate moieties tested are thought to be present on the S. mansoni sporocyst surface (mannose, galactose, fucose, N-acetyl-glucosamine, N-acetyl-galactosamine, and lactose); the other two carbohydrates tested were glucose and melibiose. ROS generation was measured using the fluorescent probe - 2',7'-dichlorofluorescein-diacetate (DCFH DA). Hemocytes were derived from two different strains of B. glabrata: one of the strains used (MO) is susceptible to infection by the trematode Schistosoma mansoni (PR-1 strain), while the other snail strain (13-16-R1) is resistant to infection with PR-1. Three of the BSA-carbohydrate conjugates (BSA-galactose, BSA mannose, and BSA-fucose), stimulated generation of reactive oxygen species in the molluscan hemocytes. The responses of the hemocytes were similar whether they were derived from susceptible or resistant snails. If the carbohydrate structures we found, to stimulate ROS generation are involved in parasite recognition, our results suggest that parasite killing may involve either qualitative differences in production of reactive oxygen species, or additional factors. PMID- 10831789 TI - Short-term immunoisolation of incompatible xenografts in a snail, Biomphalaria glabrata. AB - Hearts of the snails Physa virgata and Biomphalaria glabrata were implanted into the hemocoel of B. glabrata. Implants either were microencapsulated in 2.5% agarose or were unencapsulated. Unencapsulated xenografts from P. virgata underwent complete necrosis within 3 days post-implantation (DPI), whereas allografts were still alive at 7 DPI. In the case of microencapsulated implants, both allografts and xenografts were alive at 3 DPI, showing 4.8 and 14.9% pyknosis among myofiber nuclei, respectively. These results suggest that direct cytotoxicity by hemocytes, rather than toxic plasma factors or donor-recipient physiological incompatibility, is responsible for rapid xenograft death. However, both types of microencapsulated grafts were necrotic at 7 DPI. This necrosis may have resulted from oxygen deprivation brought about both by the agarose and by the heterotopic implantation site, inasmuch as microencapsulated allografts and xenografts cultured in saline for 7 days showed only 1.98 and 30% pyknosis, respectively. PMID- 10831790 TI - Serum amyloid A transcription in Atlantic salmon (Salmo salar L.) hepatocytes is enhanced by stimulation with macrophage factors, recombinant human IL-1 beta, IL 6 and TNF alpha or bacterial lipopolysaccharide. AB - Serum amyloid A (A-SAA) has previously been reported to be an acute-phase protein in salmonids. Hepatocytes represent a major source of A-SAA in salmonids, but nothing is known about hepatocyte SAA synthesis in fish. In the present work, the expression of A-SAA transcripts in primary cultures of Atlantic salmon hepatocytes in response to macrophage derived cytokines, human recombinant cytokines and bacterial lipopolysaccharide (LPS) was studied by Northern blot analysis. The macrophage supernatants were prepared by stimulating Atlantic salmon head kidney macrophages with LPS, yeast glucan or a leukocyte derived macrophage activating factor (MAF). The supernatants from glucan- or MAF stimulated macrophages had no effect on A-SAA expression of the hepatocytes, while supernatants from LPS-stimulated macrophages gave about a 2-fold increase in expression. The combination of either glucan and MAF, or LPS and MAF were more effective and these supernatants gave a 3.4- and 5.2-fold increase in A-SAA expression, respectively. The hepatocytes were also treated with the human recombinant cytokines TNFalpha, IL-1beta and IL-6, alone or in combination. The A SAA response to each of them alone was modest, but TNFalpha and IL-6 or IL-1beta and IL-6 in combination gave a higher response than each cytokine alone. These data suggest that the expression of A-SAA by hepatocytes from Atlantic salmon is induced by cytokine-like molecules. Interestingly, hepatocytes treated directly with LPS gave a more than 10-fold increase in SAA mRNA expression, but it is not known if this is a direct effect of LPS on the hepatocytes or if it is mediated by other contaminating cell types. PMID- 10831791 TI - Constitutive and LPS-induced gene expression in a macrophage-like cell line from the rainbow trout (Oncorhynchus mykiss). AB - The lack of cell lines from mononuclear phagocytes of salmonid fish has impeded the study of immune function at a cellular level in these economically and ecologically important animals. Here, we report on the further characterization of RTS11, a previously described macrophage-like cell line from the rainbow trout spleen, with regard to its expression of a number of immunologically relevant genes. Analysis of gene expression by reverse transcription-polymerase chain reaction, using rainbow-trout-specific primers demonstrated that RTS11 cells express the beta chain of the class II major histocompatibility complex, the cytokines transforming growth factor-beta (TGF-beta) and interleukin-1beta (IL 1beta), and cyclo-oxygenase-2 (COX-2). Inducing the cells with lipopolysaccharide led to increased expression of IL-1beta and COX-2, as determined by Northern blotting. These results together suggest that RTS11 retains many of the characteristics expected of mature macrophages, and should be a valuable tool for further study of the expression and function of these immunomodulatory proteins in fish. PMID- 10831792 TI - Factors influencing the expression of interleukin-1 beta in cultured rainbow trout (Oncorhynchus mykiss) leucocytes. AB - The dose dependency and kinetics of lipopolysaccharide (LPS) induced interleukin 1beta (IL-1beta) mRNA expression in rainbow trout leucocytes has been studied. Northern blot analysis revealed weak hybridisation of a trout IL-1beta probe to RNA from head kidney leucocytes stimulated with 0.1 microg LPS/ml, with a large increase in IL-1beta transcript level seen between 0.1 and 1.5 microg LPS/ml. Using 5 microg LPS/ml expression was first detectable 1-2 h post-stimulation. By 4 h post-stimulation maximal induction was seen but by 24-48 h the level had fallen and by 72 h no transcript was detectable. Culture temperature had a marked effect on IL-1beta expression, with low temperatures inhibiting transcription. Indeed, an eight-fold increase in IL-1beta transcript level was seen between cells cultured at 4 degrees C and 22 degrees C. Preincubation with cortisol was also shown to inhibit LPS-induced IL-1beta expression. PMID- 10831793 TI - Telomerase expression and telomere length in immortal leukocyte lines from channel catfish. AB - Normal channel catfish leukocytes readily undergo spontaneous in vitro immortalization yielding functionally active diploid cell lines. Since telomerase activation appears to be a critical step in the establishment of immortal mammalian cells, studies were undertaken to determine if and when telomerase expression occurs during the in vitro immortalization process of channel catfish leukocytes. To this end, freshly isolated peripheral blood leukocytes (PBL) from normal fish were shown to exhibit low to undetectable levels of telomerase activity and within four days after culture initiation showed dramatic increases in telomerase activity which typically remained high for at least four weeks. This activity then declined, concomitant with decreases in cellular proliferation and increases in cell death. Cells which escaped this culture "crisis" re expressed high levels of telomerase activity indefinitely. Although telomerase activity was expressed early in the immortalization process, clonal cell lines derived from these cultures had relatively short telomeres. These results suggest that telomerase expression in catfish leukocytes is activation-induced, and its expression does not necessarily stabilize telomere length until a critically, albeit ill-defined, short length is reached. PMID- 10831794 TI - A precerebellin-like protein is part of the acute phase response in rainbow trout, Oncorhynchus mykiss. AB - The acute phase response (APR) has a long evolutionary history, but it remains to be characterized fully in lower vertebrates. To study the acute phase proteins of a teleost, rainbow trout (Oncorhynchus mykiss), we induced an APR by injecting Vibrio bacterin emulsified in FIA. In samples taken over the next 3 weeks, the total plasma protein profile changed consistently as seen in one and two dimensional SDS PAGE. One 18.1 kD upregulated protein was isolated from 2D gels and an N-terminal sequence obtained. Using reverse transcriptase-PCR, a 700 bp cDNA sequence was amplified. The sequence is 53% similar at the amino acid level with rat precerebellin (regions aa 42-184 from trout and aa 89-224 precerebellin), and 46% similar with the globular portion of the human B chain of the first complement component C1q. However, it lacks the collagen portion of C1q with its characteristic Gly-X-Y repeats. The isolated protein seems to be involved in the inflammatory response but its physiological function is unknown. PMID- 10831795 TI - MHC class II A genes in the channel catfish (Ictalurus punctatus). AB - In order to characterize the Major histocompatibility complex (MHC) class II A genes of the channel catfish (Ictalurus punctatus) a cDNA library was screened and PCR was performed. Four different full-length cDNA sequences for MHC class II A genes were obtained from a clonal B cell line derived from an outbred fish. Two different genomic sequences and corresponding cDNAs were obtained from a presumably homozygous gynogenetic catfish. The A genes have five exons and four phase one introns. The first exon encodes the 5' untranslated region (UTR) and leader peptide; the second and third exons encode the alpha1 and alpha2 domains, respectively. The connecting peptide, transmembrane and cytoplasmic domains, as well as part of the 3' UTR, are encoded by the fourth exon and the rest of the 3' UTR is encoded by the fifth exon. Southern blot analyses using an exon three probe revealed two to four hybridizing fragments with considerable restriction fragment length polymorphisms evident among randomly selected outbred channel catfish. These findings are consistent with the presence of at least two functional polymorphic MHC class II A gene loci. An unusual aspect of the channel catfish MHC class II alpha chain is its lack of N-linked glycosylation sites. PMID- 10831796 TI - Requirements for nitric oxide production by turbot (Scophthalmus maximus) head kidney macrophages. AB - The effect of different cytokines and bacterial lipopolysaccharide (LPS) on turbot (Scophthalmus maximus) macrophage nitric oxide (NO) production has been studied. We have found two different responses concerning NO production in response to LPS. We have studied 43 turbot and only macrophage cultures derived from 30.2% of these turbot were significantly stimulated by LPS. The macrophage populations that did not respond to LPS, showed a constitutive production that was significantly reversed by NO inhibitors like N(G)-methyl-L-arginine (L-NMMA) and N-omega-nitro-L-arginine (L-NAME), and was dependent on intracellular calcium concentration. We studied the effect of other stimuli combined with LPS on the NO production of these otherwise non-responsive macrophages. LPS combined with turbot macrophage activating factor (MAF) containing supernatants, was capable of significantly stimulating some of these macrophage populations. The same response was observed when LPS was combined with turbot IFN-alphabeta-like substances. When LPS was combined with human recombinant tumor necrosis factor alpha (hrTNF alpha), the NO production was significantly induced in all macrophage populations studied. PMID- 10831797 TI - Specific cell-mediated cytotoxicity against a virus-infected syngeneic cell line in isogeneic ginbuna crucian carp. AB - In the present study, we attempted to determine whether virus-specific cell mediated cytotoxicity occurs in fish. Experiments were conducted with clonal ginbuna crucian carp (Carassius auratus langsdorfii) (S3n strain), and the syngeneic cell line (CFS). Two infectious viruses were used: infectious pancreatic necrosis virus (IPNV) and eel virus from America (EVA). Peripheral blood leukocytes, which consisted almost entirely (95%) of lymphocytes and thrombocytes, from S3n ginbuna immunized with virus-infected CFS cells lysed the virus-infected CFS cells (immunogen) more completely than CFS cells infected with a different virus (non-immunogen). This specific cell-mediated cytotoxicity of ginbuna against virus-infected CFS cells was efficiently induced as a result of in vivo secondary immunization. A significant specific cytotoxic activity peaked during 7-10 days after the secondary immunization. In addition, effector cells immunized with IPNV-infected syngeneic cells did not lyse IPNV-infected xenogeneic cells. These results support the hypothesis that fish exhibit specific cytotoxicity against virus-infected cells, resembling the specific cytotoxicites of higher vertebrates. PMID- 10831798 TI - Phosphatidylserine expression on apoptotic lymphocytes of Xenopus laevis, the South African clawed toad, as a signal for macrophage recognition. AB - Inflammation is avoided in apoptosis by early removal of dying cells by macrophages (MOs). In mammalian cells, an early aspect of apoptosis is the translocation of phosphatidylserine (PS) from the inner leaflet of the cell membrane to the surface. PS recognition can serve as a signal for triggering removal of dying cells. PS expression on splenocytes and thymocytes of Xenopus laevis was quantified using FITC-Annexin and flow cytometry following exposure in vitro to several known apoptogens for this species. All apoptogens used induced PS expression. Dose dependency and the kinetics of PS expression following exposure to the calcium ionophore, A23187, were also examined. Peritoneal exudate cells (PEC's) were cultured with A23187-treated thymocytes to test MO capacity for recognition of PS. MO binding to apoptotic thymocytes was reduced following exposure of PEC's to a water soluble analogue of PS, phospho-L-serine. The presence of a phagocytic PS-dependent recognition system in amphibia is supportive of the evolutionary conservation of this function in mammals that is crucial in limiting inflammation induced by dying cells. PMID- 10831799 TI - Purification of functional T lymphocytes from splenocytes of the beluga whales (Delphinapterus leucas). AB - In an effort to gain knowledge on immune functions in beluga whales, Delphinapterus leucas, we have used two physical methods for the purification of T lymphocytes of spleen cells. Isolation by sheep red blood cells (SRBC) rosetting and by adherence on nylon wool columns were tested. SRBC-rosetting gave unreliable results in obtaining purified T cells. Therefore, the purification of T cells was done using nylon wool columns. Less than 3% of the IgM(+) B cells remained in effluent populations. In the later population, 45% gave positive staining with mouse anti-human CD4 allowing us to verify functionality of the cells. The study of calcium mobilization and tyrosine kinase activation, mediated by CD4 cross-linking permitted verification of the functionality of cells. We also showed that upon activation with mitogens, beluga T cells upregulate the density of MHC class II molecules on their surfaces. CD4 cross-linking with a specific antibody inhibited the proliferation response. Overall, the activation of beluga whales lymphocytes did not differ markedly from what is known in other species. This study can help in the groundwork for functional investigation of the beluga whale's immune system. PMID- 10831800 TI - Interferon-gamma induces differentiation of ectoplacental cone cells to phenotypically distinct trophoblasts. AB - Maturation of the murine ectoplacental cone results in the development of the placental tissue which essentially consists of two trophoblastic zones, the spongiotrophoblast and labyrinthine trophoblast. In this study we attempted to investigate the action of cytokines on ectoplacental cone cell differentiation to mature trophoblast cells. After determining the cellular composition of the ectoplacental cone cell suspensions based on the expression of cytokeratin, vimentin, Mac-1, class I and class II MHC antigens, the cells were exposed to the differentiation-inducing cytokines IL-3, GM-CSF, CSF-1 and IFN-gamma. From the four factors employed, only IFN-gamma increased the levels of cytokeratin positive cells without inducing Mac-1 expression. IL-3 increased the percentages of cytokeratin as well as Mac-1- and vimentin-positive cells whereas GM-CSF and CSF-1 preferentially promoted an increase of the Mac-1 and vimentin markers. For further analysis, ectoplacental cone cells were negatively selected for Mac-1, class I and class II antigens to exclude non-trophoblastic contaminants and thereafter treated with the same cytokines. We show here that only IFN-gamma leads the sorted ectoplacental cone cells to a pure trophoblastic population composed of 100% cytokeratin-positive cells. The specificity of IFN-gamma on this differentiation pathway is strengthened by the fact that murine maternal serum on the day of EC formation contains high levels of this cytokine, suggesting that its natural presence supports - at least phenotypically - the in vivo differentiation of EC cells to trophoblasts. PMID- 10831801 TI - Changes in the development of experimental autoimmune prostatitis (EAP) by castration in aged rats. AB - During Experimental Autoimmune Prostatitis (EAP), 12-month-old rats show a higher cellular autoimmune response and lower humoral autoimmune response against prostatic components than 3-month-old rats subjected to the same antigen stimulus. We analyzed if thymus recovery by orchidectomy could affect the development of EAP in 12-month-old rats. Thirty days after gonadectomy, 12-month old rats showed an increment in the thymic mass and in the thymocytes absolute number, with percentages of the four main cell subpopulations (defined by CD4-CD8 molecules expression) similar to the 3-month-old rats. The DTH response of castrated 12-month-old with EAP were diminished in comparison with sham-castrated 12-month-old rats with EAP, resembling the values observed in 3-month-old rats with EAP. The prostates of castrated 12-month-old rats with EAP did not show inflammatory mononuclear cell infiltration, as did control 3- and 12-month-old rats with EAP. Castration seems to modulate negatively EAP in 12-month-old rats, possibly through the regeneration of thymus after testosterone deprivation. PMID- 10831802 TI - Developmental study of immunocompetent cells in the bronchus-associated lymphoid tissue (BALT) from Wistar rats. AB - The aim of the present report was to study in growing Wistar rats the development of immunocompetent cells in the bronchus-associated lymphoid tissue (BALT). We found at day 4 postpartum, a high number of TCRgamma/delta+ T cells and very few CD8alpha+, CD8beta+, CD5+, TCRalpha/beta+ T cells in BALT. The latter cells and CD4+ T cells increase with age. Even though T cells expressing TCRgamma/delta outnumber those expressing TCRalpha/beta early in development, until 45 days of age, alpha/beta+ predominate over gamma/delta+ T cells only in adult rats (60 days of age). Moreover, a predominance of suppressor/cytotoxic T cells over T helper cells was found in 60 days old rats. Surprisingly, more CD8alpha+ than CD8beta+ T cells in BALT are observed. The number of IgA+ B and CD4+ T cells found in the BALT increases with age. The early appearance - 4 days of age - of all T-cell phenotypes in BALT especially of gamma/delta+ T cells may imply a benefit to respond to inhaled antigen soon after birth. PMID- 10831803 TI - An enhanced role for the recirculating lymphocyte in the neonatal immune system. AB - Lymphocytes continually recirculate between the blood and the tissues via the lymph independent of antigen. A great deal is known regarding both the physiology and the molecular mechanisms responsible for the process in adults. However, relatively little is known regarding the development of the recirculating lymphocyte pool in very young animals or fetuses. We have directly measured the recirculation of lymphocyte subsets in antigen-inexperienced newborn animals, and found extensive recirculation of T cells through both intestinal and subcutaneous lymph nodes. Apparent selective migration of recirculating lymphocytes could be attributed to subset-specific migration of gammadelta-T cells through subcutaneous lymph nodes. This clearly demonstrates that the preference for gammadelta-T cells to recirculate through SCLN is lineage specific, and independent of the presence of antigen. Most surprising was the observation that the recirculating lymphocyte pool was proportionately larger in neonatal animals than in adults, which correlated with the histological appearance of newborn lymph nodes. This data strongly suggests that development of the recirculating lymphocyte pool is inversely correlated with antigen exposure, and decreases in size with age and the acquisition of immunological memory. PMID- 10831805 TI - L.C. Smith, C. baier-anderson, L.A. Clow, D.P. Terwilliger and C.M. Adema. Conference report. Developmental and comparative immunology 24 (2000) 71--77 PMID- 10831804 TI - Characterization of expanded T cell clones in healthy macaques: ontogeny, distribution and stability. AB - Peripheral expanded T cell clones have been discussed mainly in relation to certain diseases or immune function in humans and mice. There is little information on their ontogeny, stability and distribution among T cell subsets as well as major lymphoid organs. We applied reverse transcription-polymerase chain reaction (RT-PCR) with family specific primers for monkey T cell receptor beta chain V regions and single-strand conformation polymorphism (SSCP) analysis to analyze the expanded T cell clones in cynomolgus monkeys (Macaca fascicularis). A number of expanded T cell clones were detected in the peripheral blood of young and adult monkeys, but few expanded T cell clones were detected in the blood of a fetus and a 2-day-old neonate. The clones in adults were maintained over 3 months. These expanded T cell clones were distributed only in peripheral blood and spleen, but few were found in lymph nodes (axillary, inguinal and intestinal). The number of expanded T cell clones was much greater in CD8 single positive (CD8sp) T cells than in CD4sp T cells, showing that most of these clones originated in the CD8sp T cell population. Almost all the expanded CD8sp T cell clones belonged to the CD28(-), CD29(hi) and Fas(+) subset. The usage of V beta genes was not skewed in the 24 V beta. Furthermore, higher mRNA signals for effector molecules perforin and IFN-gamma were detected in CD8sp T cell subsets with phenotypes of CD28(-), CD29(hi) and Fas(+), suggesting that the expanded T cells might have developed in relation to T cell activation in the periphery of cynomolgus monkeys. PMID- 10831806 TI - Lifting an unexpectedly heavy object: the effects on low-back loading and balance loss. AB - OBJECTIVE: This study evaluates the effects of lifting an unexpectedly heavy object on low-back loading and loss of balance. BACKGROUND: It is often suggested that lifting an unexpectedly heavy object may be a major risk factor for low-back pain. This may lead to an increase in muscle activation, stretch of ligaments and posterior disc, and loss of balance.METHODS. Nine healthy male subjects were asked to pick up and lift a box as quickly as possible. The weight of the box was unexpectedly increased by 5 or 10 kg. Kinematics and force data were recorded throughout the experiment. RESULTS: Lifting of an unexpectedly heavy box led to a decrease in maximum torque of the low back compared to lifting the same box mass with correct expectation. The maximum lumbar angle did not increase compared to the light box condition. Only the threat to balance appeared to be somewhat increased.CONCLUSIONS. The lifting of an unexpectedly heavier box appeared not to lead to an increased balance loss or a clearly increased stress of the structures of the low back, although a burst of abdominal muscle activity was found in one condition. These results do not fully clarify the assumed relation between lifting unexpectedly heavy objects and low-back injury. RelevanceA commonly cited cause of low-back pain is the lifting of an unexpectedly heavy object. A study of the responses to such perturbation is important to an understanding of spine mechanics and the etiology of low-back injury. PMID- 10831807 TI - Strategies of load tilts and shoulders positioning in asymmetrical lifting. A concomitant evaluation of the reference systems of axes. AB - OBJECTIVES: To evaluate two handling factors on asymmetry of posture and efforts and to evaluate different reference systems of axes on the characterization of asymmetry.DESIGN. Ten novice workers performed three tasks evaluated for the effects of load tilting (tilts vs no tilts) and shoulders positioning (non parallel vs parallel to the ground). Specific comparisons were made using three referential systems.BACKGROUND. Box handgrips/tilting and body posture are factors differentiating expert and novice workers which present a potential for reducing asymmetries. The literature also suggests that different conclusions may be reached about asymmetry with different systems of axes.METHODS. Net 3D moments at L5/S1 were obtained from two force platforms, four video cameras and inverse dynamic analyses using three reference systems of axes (trunk, pelvis, and joint).RESULTS. Tilting the load presented clear advantages of reducing the duration and trajectory of efforts, better positioning the load and reducing knee flexion, peak trunk extensors and mechanical work on the load. The maintenance of the shoulders parallel reduced asymmetries of posture and efforts but the results were affected by the type of reference system of axes chosen.CONCLUSIONS. Box tilting and shoulders positioning should be considered in the reduction of risk factors. More research is needed to define referential systems and characterize asymmetry. RelevanceBox tilting and shoulders positioning appear to be two fundamental elements to take into account in the formation programs based on prevention of risk factors. The importance of asymmetry among the risk factors emphasizes the need for better characterization of this risk. PMID- 10831808 TI - Finger motion, wrist motion and tendon travel as a function of keyboard angles. AB - OBJECTIVE: This study assessed the impact of keyboard angles (in terms of Pitch, Roll and Yaw) on tendon travel and wrist and finger joint kinematics for the flexor digitorum profundus and flexor digitorum superficialis.DESIGN. A repeated measures, laboratory study was conducted. Independent variables were three Pitch angles, three Roll angles, three Yaw angles, and three keyboard separation distances. Dependent variables were tendon travel, wrist deviation, wrist and finger joint kinematics, and Borg comfort rating. BACKGROUND: The increased usage of computers and the risk of cumulative trauma disorders have led to the development of alternate keyboards. This study is a biomechanical assessment of several keyboard designs.METHODS. Lightweight wrist and finger goniometers were used to measure motion of the wrist in three planes, and for three finger joints. Fifteen experienced typists (eight women, seven men) typed a standard text on 30 keyboard conditions. Regression equations were used to calculate tendon travel from joint positions. RESULTS: Tendon travel is sensitive to changes in Pitch, Roll and Yaw angles with approximately 13% difference between the minimal and maximal tendon travel. A flat keyboard produced more tendon travel than keyboards with greater Pitch and Roll angles.CONCLUSIONS. There is a trade-off between wrist and finger positions; as the wrist extends more, the finger joints flex more to compensate. Keyboards imposed different trade-offs between the wrist and finger positions, affecting the overall tendon travel. Relevance Alternate keyboard designs can significantly affect tendon travel and may address reduced repetitiveness in typing by reducing the amount of tendon travel. PMID- 10831809 TI - An assessment of alternate keyboards using finger motion, wrist motion and tendon travel. AB - OBJECTIVE: To assess the biomechanical impact of commercially available alternate keyboard designs.DESIGN. A repeated measures study was conducted in a laboratory setting, with planned comparisons of Pitch, Roll and Yaw angles of the keyboards. Ten keyboard conditions were tested. Dependent measures included tendon travel, wrist deviations, and wrist and finger kinematics.BACKGROUND. Various alternate keyboard designs have recently been introduced, which vary Pitch, Roll and Yaw angles, separation distance between keyboard halves, and include other novel features such as cup-shaped depressions for the keys. Yet little objective research has been conducted regarding the biomechanical implications of these various design features. This study attempts to quantify the keyboard designs in terms of several recognized risk factors associated with cumulative trauma disorders that arise with repetitive typing. METHODS: Wrist and finger goniometers were used to measure joint motions during a standardized typing task. 15 experienced typists (8 women, 7 men) served as subjects. Regression equations were used to generate estimates of tendon travel.RESULTS. Tendon travel was affected primarily by Pitch but not Roll or Yaw angles while wrist deviations responded to changes in all three angles. Males had significantly greater amount of tendon travel than female subjects; this difference was only partially accounted for by anthropometry. Differences in joint motion may have a greater impact on the amount of tendon travel.CONCLUSIONS. Alternate keyboard designs can affect tendon travel by as much as 11%. RelevanceAs various alternate keyboard designs are marketed, quantifiable biomechanical data such as that provided by this study, will help to assess their impact on the risk factors for cumulative trauma disorders. PMID- 10831810 TI - Three-dimensional gait analysis in women with a total hip arthroplasty. AB - OBJECTIVES: The purposes of this study were: (1) to identify the primary (hip) and secondary (neighboring joints) impairments during gait in subjects with a total hip arthroplasty total hip arthroplasty, (2) to determine which impairments persist when controlling for gait speed and (3) to study the relationships between primary and secondary impairments in order to describe the locomotor strategies used by these patients.DESIGN. This cross-sectional study compared the gait patterns of women with a total hip arthroplasty to those of healthy women.BACKGROUND. Several studies have reported residual hip impairments in the sagittal plane during gait after a total hip arthroplasty. There is, however, a substantial lack of knowledge in regard to the changes at the neighboring joints and in the other planes of movement.METHODS. Subjects have been examined during a gait laboratory testing session including the simultaneous recording of three dimensional kinematics and ground reaction forces on one side, and bilateral activation of six lower limb muscles.RESULTS. A significant decrease of 20% in the hip extensor moment of force during the early stance phase was correlated (R(2)=43%) with a significant decrease of 14% in gait speed. Moreover, a significant decrease of 59% in the range of hip extension at the end of the stance phase was observed together with secondary impairments such as a significant increase in the anterior pelvic rotation, in knee flexion and in ankle dorsiflexion. Lastly, a significant increase in ipsilateral bending of the trunk during the single limb support on the operated limb was concomitant with a significant decrease in the hip abductor moment of force.CONCLUSIONS. The decrease in gait speed and the persistence of abnormal gait patterns one year after the total hip arthroplasty were associated respectively with a decrease in the hip extensor moment of force and with a decrease in the range of hip extension (sagittal plane) or in the hip abductor moment of force (frontal plane). RelevanceThe data provided in this paper may serve as guidelines to establish rehabilitation programs designed to restore optimal locomotor function. PMID- 10831811 TI - Impact load transmission of the knee joint-influence of leg alignment and the role of meniscus and articular cartilage. AB - OBJECTIVE: The objective of this study was to evaluate load transmission in the cancellous bone of the tibia under static and impact load.BACKGROUND. Abnormal transmission of loads may result in osteoarthritis and fractures. However, the role of the cancellous bone in these processes is not well understood.METHODS. The compressive stresses in the subchondral bone, epiphysis and diaphysis of the tibia of porcine knees were measured under static and impact load using mini pressure transducers. The tests were performed using a drop-tower type testing machine in neutral, varus and valgus alignments. Tests were repeated after meniscectomy and again after removing the articular cartilage.RESULTS. In the intact knee in all alignments, the highest stress on the medial side was found in the epiphysis, and in the subchondral bone on the lateral side. After meniscectomy, a significant increase was observed in the stress in the subchondral bone on both sides. After the articular cartilage was also removed, the stress in the subchondral bone increased again, but slightly.CONCLUSION. The importance of the meniscus in load transmission is supported by this study. The contribution of the articular cartilage to load transmission is less than that of the meniscus.Relevance. The patterns of static and dynamic load transmission observed in this study correlate with the sites of insufficiency fractures of the medial tibial plateau and traumatic lateral tibial plateau fractures observed clinically. Examination of the load transmission is key to a more complete understanding of the causes of osteoarthritis and tibial plateau fractures. PMID- 10831812 TI - The relationship between ankle plantar flexor muscle moments and knee compressive forces in subjects with and without pain. AB - OBJECTIVE: The purpose of this study was to compare the compressive knee joint reaction force and ankle plantar flexor muscle moment during the terminal stance phase in symptomatic osteoarthritic and non-symptomatic subjects.DESIGN. The study design was a comparative investigation of walking patterns.BACKGROUND. Patients with knee osteoarthritic symptoms may employ altered gait patterns to reduce the loading of painful joints. Speculation is that decreased ankle plantar flexor moments in the terminal stance phase may be an attempt to minimize compressive knee joint reaction forces.METHODS. Twenty community-dwelling men (52 83 years old), ten with knee osteoarthritis and ten non-symptomatic, were studied while walking across a walkway containing two force plates at a comfortable speed. Retroreflective markers were placed in accordance with the Helen Hayes marker system and recorded with six 120 Hz cameras. Three-dimensional kinematics and kinetics were computed.RESULTS. Group means for gait velocity, peak ankle plantar flexor moment and compressive knee joint reaction force were significantly less for the osteoarthritic group relative to the non-symptomatic group. When accounting for the decreased velocity in the osteoarthritic group, however, no group differences were observed. There were significant correlations between all three gait parameters (r>0.75), but peak ankle plantar flexor moment was the greatest predictor of compressive knee joint reaction force.CONCLUSIONS. While no group differences were found for compressive knee joint reaction forces when accounting for gait velocity, it appears that subjects with osteoarthritis utilize gait velocity as a mechanism to reduce compressive knee joint reaction forces. RelevanceAn understanding of compressive knee joint reaction forces is important to the understanding of the disabling effects of knee osteoarthritis on physical function and to the understanding of appropriate and safe interventions to improve not only global parameters (e.g., pain, gait velocity), but also to "re-program" the locomotor pattern to develop "normal" muscle moments without increasing compressive knee joint reaction forces to painful levels. PMID- 10831813 TI - Knee joint loading in forward versus backward pedaling: implications for rehabilitation strategies. AB - OBJECTIVE: To use forward dynamic simulations of forward and backward pedaling in order to determine whether backward pedaling offers theoretical advantages over forward pedaling to rehabilitate common knee disorders.DESIGN. A comparison of knee joint loads was performed during forward and backward pedaling.BACKGROUND. Pedaling has been shown to be an effective rehabilitation exercise for a variety of knee disorders. Recently, backward gait has been shown to produce greater knee extensor moments and reduced patellofemoral joint loads compared to forward gait. But to date, no study has examined the efficacy of backward pedaling as a safe alternative to forward pedaling in rehabilitation programs.METHODS. A musculoskeletal model and optimization framework was used to generate simulations of forward and backward pedaling. Tibiofemoral and patellofemoral joint reaction forces were quantified.RESULTS. Lower tibiofemoral compressive loads, but higher patellofemoral compressive loads, were observed in backward pedaling. Lower protective anterior-posterior shear force was observed in backward pedaling near peak extension.CONCLUSIONS. Backward pedaling offers reduced tibiofemoral compressive loads for those patients with knee disorders such as menisci damage and osteoarthritis, but higher patellofemoral compressive loads. Therefore, backward pedaling is not recommended for patients experiencing patellofemoral pain. Further, backward pedaling should not be recommended after anterior cruciate ligament injury or reconstruction.RelevanceThe results of this study indicate that the design of rehabilitation programs including pedaling exercises should be injury specific with particular attention paid to the mechanics of the task. PMID- 10831814 TI - The effects of knee and ankle flexion on ground clearance in paraplegic gait. AB - OBJECTIVE: To investigate the effects of knee flexion and ankle dorsiflexion on ground clearance during the swing phase of paraplegic gait.BACKGROUND. Limitations of currently available walking orthoses are high energy consumption and upper limb loading. Knee flexion in the swing phase has been suggested as a means of reducing these.METHODS. Kinematic data of paraplegic gait in two currently available orthoses were used to simulate the effects of knee flexion and ankle dorsiflexion on the swing foot ground clearance. This was used to validate a mathematical model of a simple ideal gait that was also developed.RESULTS. It was shown that the implementation of knee flexion alone leads to a loss or no change in ground clearance. Implementing knee flexion and ankle dorsiflexion can be used to increase ground clearance.CONCLUSION. Flexion at the knee and ankle during the swing phase can potentially be used to allow a reduction in compensatory mechanisms by easing swing foot ground clearance. RelevanceWalking orthoses are valuable in the clinical management of paraplegia, providing both physiological and psychological benefits. Functional improvements can only serve to encourage their greater usage. PMID- 10831815 TI - Amplitude and frequency analysis of force plate data in sitting children with and without MMC. AB - OBJECTIVE: To investigate if force plate measurements can be used to detect postural sway differences in sitting children with and without myelomeningocele (spina bifida).BACKGROUND. The postural sway has not been investigated in children with myelomeningocele previously. Since many of these children are not able to stand independently, force plate measurements during sitting could be one way to detect differences in their postural sway compared to normal children. However, there is very little published regarding assessment of seated postural sway.METHODS. Force plate measurements on 15 six years old children with myelomeningocele and 20 age-matched normal children were analysed. The standard deviation and the median frequency of the horizontal ground reaction force were used to characterise the body sway.RESULTS. The standard deviation of the force was larger only for some of the children with myelomeningocele as compared to the control group. The median frequency was significantly lower in the myelomeningocele group as compared to the control group. Visual input and seat base inclination did not influence the postural sway significantly. CONCLUSIONS: The results show that frequency analysis can be used to detect fundamental differences in postural sway that can not be observed visually. RelevanceIn this paper a new method for analysis of seated postural sway is described. The lack of relatively high spectral frequencies for children with myelomeningocele shows that the output from the postural control system differs as compared to the control group. PMID- 10831816 TI - Stadiometry: on measurement technique to reduce variability in spine shrinkage measurement. AB - OBJECTIVE: To test the effect of two measurement techniques for repeated measures of spine height using stadiometry following five experimental activity conditions.DESIGN. Six subjects were repeatedly measured while they stepped in and out of the stadiometer for each pair of measures and again on another day when they remained in place in the stadiometer for all 10 measures.RESULTS. There was much greater variability in height measures with the "in-out" method while the "in place" method demonstrated a steady shrinkage over the 3-3.5 min required to obtain the repeated measures. RelevanceContrary to popular practice, leaving a subject in the stadiometer during repeated measures includes the shrinkage that occurs over the 3-3.5 min of measurement when standing and reduces random variation due to posture change. PMID- 10831834 TI - Groucho/TLE family proteins and transcriptional repression. AB - The Drosophila Groucho (Gro) protein is the prototype for a large family of corepressors, examples of which are found in most metazoans. This family includes the human transducin-like Enhancer of split (TLE) proteins. As corepressors, Gro/TLE family proteins do not bind to DNA directly, but rather are recruited to the template by DNA-bound repressor proteins. Gro/TLE family proteins are required for many developmental processes, including lateral inhibition, segmentation, sex determination, dorsal/ventral pattern formation, terminal pattern formation, and eye development. These proteins are characterized by a conserved N-terminal glutamine-rich domain and a conserved C-terminal WD-repeat domain. The primary role of the glutamine-rich domain is apparently to mediate tetramerization, while the WD-repeat domain may mediate interactions with DNA bound repressors. The glutamine rich and WD-repeat domains are separated by a less conserved region containing domains that have been implicated in transcriptional repression and nuclear localization. In addition to encoding full length Gro/TLE family proteins, most metazoan genomes encode truncated family members that contain the N-terminal oligomerization domain, but lack the C terminal WD-repeat domain. These truncated proteins may negatively regulate full length Gro/TLE proteins, perhaps by sequestering them in non-productive complexes. Gro/TLE family proteins probably repress transcription by multiple mechanisms. For example, a glycine/proline-rich domain in the central variable region functions to recruit the histone deacetylase Rpd3 to the template. This histone deacetylase then presumably silences transcription by altering local chromatin structure. Other repression domains in Gro may function in a histone deacetylase-independent manner. Many aspects of Gro/TLE protein function remain to be explored, including the possible post-translational regulation of Gro/TLE activity as well as the mechanisms by which Gro/TLE proteins direct repression at a distance. PMID- 10831835 TI - Transforming growth factor beta signaling mediators and modulators. AB - Transforming growth factor beta is a multi-functional growth and differentiation factor responsible for regulating many diverse biological processes in both vertebrate and invertebrate species. Among the most dramatic of TGFbeta's effects are those associated with specification of cell fates during development and inhibition of cell cycle progression. The core TGFbeta signaling pathway has now been described using a synergistic combination of genetic and biochemical approaches. Transmembrane receptors with intrinsic protein serine kinase activity bind ligand in the extracellular milieu and then phosphorylate intracellular proteins known as Smads. Phosphorylated Smads form heterooligomers and translocate into the nucleus where they can modulate transcriptional responses. More recent studies indicate that many other proteins serve as modulators of Smad activity, and utimately define specific cellular responses to TGFbeta. Here we describe both the simplistic core TGFbeta signaling pathway and the growing number of proteins that impinge on this pathway at the level of Smad function to either enhance or inhibit TGFbeta responses. PMID- 10831836 TI - Roles of the NFI/CTF gene family in transcription and development. AB - The Nuclear Factor I (NFI) family of site-specific DNA-binding proteins (also known as CTF or CAAT box transcription factor) functions both in viral DNA replication and in the regulation of gene expression. The classes of genes whose expression is modulated by NFI include those that are ubiquitously expressed, as well as those that are hormonally, nutritionally, and developmentally regulated. The NFI family is composed of four members in vertebrates (NFI-A, NFI-B, NFI-C and NFI-X), and the four NFI genes are expressed in unique, but overlapping, patterns during mouse embryogenesis and in the adult. Transcripts of each NFI gene are differentially spliced, yielding as many as nine distinct proteins from a single gene. Products of the four NFI genes differ in their abilities to either activate or repress transcription, likely through fundamentally different mechanisms. Here, we will review the properties of the NFI genes and proteins and their known functions in gene expression and development. PMID- 10831837 TI - Isolation of binary species-specific PCR-based markers and their value for diagnostic applications. AB - Representational difference analysis (RDA), a technique for the isolation of differences between highly similar complex genomes, was employed for isolation of species-specific markers. These markers can be easily adapted for a high throughput PCR-based assay in which multiple specimens can be simultaneously identified based on the presence/absence of amplification products. One of the important features of RDA performed on genomes of different species (interspecific RDA) is its ability to preferentially isolate families of repetitive sequences that are unique to one of the compared genomes. Such families of repetitive DNA are homoplasy-free characters that can be used for cost-efficient, mass identification of specimens in a variety of situations ranging from mark-recapture studies to screenings of egg or larval stages. PMID- 10831838 TI - GP73, a novel Golgi-localized protein upregulated by viral infection. AB - We report the isolation and characterization of GP73, a novel 73kDa human Golgi protein. The GP73 cDNA was cloned by differential screening of a cDNA library derived from the liver of a patient with adult giant-cell hepatitis (GCH), a rare form of hepatitis with presumed viral etiology. In vitro transcription translation studies indicate that GP73 is an integral membrane protein, and immunolocalization experiments using epitope-tagged GP73 demonstrate that the protein is localized to the Golgi apparatus. Northern blot analysis of RNA from multiple human tissues reveals a single GP73 mRNA transcript with a size of approximately 3.0kb. Immunohistochemical studies using rabbit polyclonal antisera directed against recombinant GP73 demonstrate that the protein is preferentially expressed by epithelial cells in many human tissues. In normal livers, GP73 is consistently present in biliary epithelial cells, whereas hepatocytes show little or no signal. In contrast, livers of patients with GCH display strong GP73 immunoreactivity in multinucleated hepatocytes. GP73 mRNA and protein are expressed in highly differentiated HepG2 hepatoma cells after infection with adenovirus in vitro. We conclude that GP73 represents a novel, epithelial cell specific integral membrane Golgi protein that can be upregulated in response to viral infection. PMID- 10831839 TI - Molecular and cellular characterizations of a cDNA clone encoding a novel isozyme of aldehyde dehydrogenase from rice. AB - Aldehyde dehydrogenases (ALDHs) are a group of enzymes catalyzing the conversion of aldehydes to the corresponding acids. In mammals and yeasts, at least two isozymes of ALDH are known to be involved in ethanol metabolism (cytosolic ALDH1 and mitochondrial ALDH2). Although mitochondrial ALDH isozymes have previously been identified in several plants, such as maize and tobacco, it is unclear whether cytosolic ALDH isozymes also exist in plants. In this study, we identified and characterized a cDNA clone encoding aldehyde dehydrogenase (ALDH1a) from rice (Oryza sativa L. cv. Nipponbare). The open reading frame of this clone did not contain a typical mitochondrial targeting signal. Analysis of the subcellular localization of ALDH1a using green fluorescent protein (GFP) suggested that ALDH1a is a cytosolic enzyme rather than a mitochondrial enzyme. A genomic Southern hybridization indicated that sequences homologous to the ALDH1a gene are present in at least two regions of the rice genome. Amplification by RT PCR showed that ALDH1a is expressed strongly in roots, but not in leaves, of rice seedlings, suggesting that ALDH1a functions in roots. PMID- 10831840 TI - Primary structure and expression of the xiphophorus DNA-(cytosine-5) methyltransferase XDNMT-1. AB - Small aquarium fishes become increasingly important in the study of normal vertebrate development and disease. Differential DNA methylation might play a role in these processes. In the teleost Xiphophorus, a well-established animal model for melanoma formation, tumour-specific hypomethylation of the melanoma inducing gene ONC-Xmrk has been observed. We have isolated a cDNA for the DNA (cytosine-5)-methyltransferase XDNMT-1 from this organism, which encodes the first full-length protein from a fish species. Linkage analysis showed that Xdnmt 1 is different from the Xiphophorus tumour suppressor R, which is involved in the transcriptional repression of the ONC-Xmrk melanoma oncogene in healthy fish. As methylation has been implicated in the regulation of ONC-Xmrk expression, XDNMT-1 might play a role by acting up- or downstream of R. Expression analysis demonstrated that the Xdnmt-1 transcript is present in all adult tissues and cell lines tested. However, developing embryos show a spatially and temporally regulated expression pattern suggesting that the enzyme might play a role during development in fish. PMID- 10831841 TI - Cloning of regulatory sequences mediating guard-cell-specific gene expression. AB - This report describes the use of promoter trap lines from the model plant Arabidopsis thaliana to clone regulatory sequences that mediate guard-cell specific reporter gene expression. Stomatal guard cells represent a highly differentiated cell type within the epidermis of green tissues of higher plants. They control the stomatal aperture in response to different endogenous and environmental signals in order to optimize carbon fixation while minimizing water loss. We screened available promoter trap lines for guard-cell-specific activation of a beta-glucuronidase (uidA) reporter gene in order to obtain marker lines for guard-cell development and to gain access to regulatory pathways leading to gene expression which is restricted to this cell type. From two lines identified we successfully cloned upstream regulatory sequences. For one line, guard-cell-specific promoter activity was confirmed by re-introducing the uidA gene, fused to the newly identified regulatory sequences, into the Arabidopsis nuclear genome. However, DNA sequences downstream of the uidA/T-DNA insertion sites in the original promoter trap lines revealed no obvious coding regions in the corresponding orientation, indicating that we have probably identified 'cryptic' promoters, being active in guard cells. PMID- 10831842 TI - Molecular cloning, chromosome mapping and characterization of UBQLN3 a testis specific gene that contains an ubiquitin-like domain. AB - The sequence of the ubiquitin protein is highly conserved between species and has facilitated the cloning of numerous ubiquitin-like proteins. In the present study, we report the cloning of the cDNA for human ubiquilin 3 (UBQLN3). The deduced amino acid sequence of UBQLN3 contains a UBQ domain (ubiquitin-like) in the amino terminus as well as two highly conserved domains found in several recently cloned ubiquitin-like proteins. One of these domains, termed the NP domain, is a highly conserved 93 amino acid region present in UBQLN3 and several ubiquitin-like proteins. The last conserved domain is the UBA domain (ubiquitin associated) found in a variety of proteins of the ubiquination pathway. The human UBQLN3 gene was mapped to the 11p15 region of chromosome 11. Northern blot analysis of multiple human and mouse tissues demonstrated UBQLN3 mRNA expression specifically in testis. PMID- 10831843 TI - Genomic structure and chromosomal mapping of the mouse hic-5 gene that encodes a focal adhesion protein. AB - The hic-5 gene encodes a focal adhesion protein that has striking similarity to paxillin. Genomic clones of the mouse hic-5 gene were isolated, and included 10 exons that covered the whole mouse mRNA sequence. Comparison of the sequence with those in the expressed sequence tag database suggested that the hic-5 gene contained an extra exon (named exon 1') located about 1kb upstream of exon 1, and mouse cells seemed to express two alternatively spliced forms of mRNA. All the exon-intron boundaries followed the GT/AG rule. Physical mapping and fluorescent in situ hybridization analysis indicated that the hic-5 gene is located on mouse chromosome 7, 60. 0cM from the centromere. PMID- 10831844 TI - A new family of genes and pseudogenes potentially expressing testis- and brain specific leucine zipper proteins in man and mouse. AB - We have characterized a new mouse gene highly transcribed in the testis, and a derived intronless gene expressed in the embryo. The latter gene is present in Mus musculus domesticus and in Mus musculus castaneus but is absent in Mus spretus. The sequencing of different clones from a testis cDNA library reveals a complex transcriptional regulation for the intron-containing gene. The use of several promoters, alternative splicing and trans-splicing, and of two different polyadenylation sites account for the diversity. The different cDNAs encode proteins with features of basic helix-loop-helix leucine zipper (bHLH-ZIP) DNA binding factors with homology to a new brain-specific factor. The presence of multiple CK2 and PKC phosphorylation sites suggests that their activity may be regulated by phosphorylation. In man, a pseudogene, apparently derived from the same transcript as in mouse and showing 90% homology in the coding region, is present within an intron of another gene. Interestingly, although the human pseudogene is highly mutated in human, in the mouse it has only four nucleotide changes compared with the cDNA of origin, and is still capable of encoding a protein. PMID- 10831845 TI - Comparative analysis of intra-individual and inter-species DNA sequence variation in salmonid ribosomal DNA cistrons. AB - This study examines sequence divergence in three spacer regions of the ribosomal DNA (rDNA) cistron, to test the hypothesis of unequal mutation rates. Portions of two transcribed spacers (ITS-1 and 5' ETS) and the non-transcribed spacer (NTS) or intergenic spacer (IGS) formed the basis of comparative analyses. Sequence divergence was measured both within an individual lake trout (Salvelinus namaycush) and among several related salmonid species (lake trout; brook trout, Salvelinus fontinalis; Arctic char, Salvelinus alpinus; Atlantic salmon, Salmo salar; and brown trout, Salmo trutta). Despite major differences in the length of the rDNA cistron within individual lake trout, minimal sequence difference was detected among cistrons. Interspecies comparisons found that molecular variation in the rDNA spacers did not conform to the predicted pattern of evolution (ITS spacers100 DPS), increases (>10 dB) in threshold level were observed for 100 Hz sinusoids in three of 11 cases. These changes were transient in one case and long-term in two cases. The time course of threshold change, both early and late, could not be explained on the basis of changes in spiral ganglion cell survival. The guinea pig seems to be an ideal preparation for studies of this nature, because threshold changes are similar in type, but accelerated in time course, relative to those observed in primates. PMID- 10831877 TI - Cochlear de-efferentation and impulse noise-induced acoustic trauma in the chinchilla. AB - The olivocochlear bundle (OCB) has been shown to protect the ear from acoustic trauma induced by continuous noise or tones. The present study examines the OCB's role in the ear's response to impulse noise (150 dB pSPL, 100 impulses, 50 s total exposure duration). Successful section of the OCB was achieved through a posterior parafloccular fossa approach for the right ears of six out of 15 adult chinchillas. The left ears from the same animals served as efferent-innervated controls. Measurements of inferior colliculus evoked potentials (ICPs) showed that the de-efferented ears incurred similar temporary and permanent threshold shifts as the control ears. Twenty days after noise exposure, depressed ICP amplitudes had virtually recovered to pre-values in the control ears whereas those in the de-efferented ears remained significantly depressed. Greater loss of inner hair cells was seen in the de-efferented ears than in the control ears. Both control and de-efferented ears incurred large loss of outer hair cells, with no statistically significant differences between groups. The current data are intriguing, yielding tentative evidence to suggest that inner hair cells of de efferented ears are more susceptible to impulse noise than those in efferented control ears. In contrast, outer hair cell vulnerability to impulse noise appears to be unaffected by de-efferentation. PMID- 10831878 TI - Hearing loss following exposure during development to polychlorinated biphenyls: a cochlear site of action. AB - Maternal exposure to polyhalogenated hydrocarbons results in early postnatal hypothyroxenemia and a low-frequency hearing loss in adult offspring (Goldey et al., 1995a. Toxicol. Appl. Pharmacol. 135, 67-76; Herr et al., 1996. Fundam. Appl. Toxicol. 33, 120-128). The purpose of the present work was to determine whether the site-of-action of this auditory impairment was within the cochlea. Primiparous Long-Evans rats were given daily oral doses of corn oil (control) or 8 mg/kg of the commercial PCB mixture Aroclor 1254 (A1254) from gestation day (GD) 6 through postnatal day (PND) 21. Auditory thresholds for 1-, 4-, 16-, and 40-kHz tones were assessed using reflex modification audiometry in young adult offspring on postnatal days (PND) 92-110. Approximately 6 weeks after auditory assessments, a subset of animals (n=4 per group) were killed for histological assessment of the cochlea. Surface preparations of the organ of Corti were prepared from one cochlea per animal and modiolar sections were prepared from the opposite cochlea. Consistent with previous findings, auditory thresholds for 1 kHz tones were elevated by approximately 25 dB in the A1254-exposed animals. Thresholds for all higher frequencies were not different compared to controls. Surface preparations of the organ of Corti revealed a mild to moderate loss of outer hair cells in the upper-middle and apical turns. Inner hair cells were not affected. Modiolar sections failed to reveal alterations in any other cochlear structures. There was also no apparent loss of ganglion cells. These data clearly link the loss of low-frequency hearing caused by exposure during development to A1254 to a loss of outer hair cells in the organ of Corti. The mechanism that underlies this developmental ototoxicity remains to be determined. These data provide the first evidence of a structural deficit in the nervous system of adult animals exposed to PCBs during development. PMID- 10831879 TI - Retreating residual refractive errors after excimer surgery of the cornea: PRK versus LASIK. PMID- 10831880 TI - New terminology: ophthalmic viscosurgical devices. PMID- 10831881 TI - Initial and definitive capsulorhexes: an extended application. PMID- 10831882 TI - In the article "Cataract Surgery in the Very Elderly" (J Cataract Refrac Surg 2000; 26:408-414), Table 4, page 411, contains a mistake. The table should be as follows: PMID- 10831883 TI - Consultation. PMID- 10831884 TI - Consultation section. Cataract surgical problem. PMID- 10831885 TI - Consultation. PMID- 10831887 TI - Consultation. PMID- 10831886 TI - Consultation. PMID- 10831888 TI - Consultation. PMID- 10831889 TI - Consultation. PMID- 10831890 TI - Consultation. PMID- 10831891 TI - Two-compartment technique to remove ophthalmic viscosurgical devices. AB - To avoid postoperative intraocular pressure peaks, complete removal of all ophthalmic viscosurgical devices (OVDs) used during cataract surgery is important. Depending on the kind of OVD used, different removal techniques have been applied. We describe a technique that was used in more than 250 eyes for safe removal of OVDs, especially viscoadaptive OVDs such as sodium hyaluronate 2.3% (Healon5). It takes advantage of the viscoadaptive properties of Healon5 and was named the 2-compartment technique. PMID- 10831892 TI - Endoscopic technique for suturing posterior chamber intraocular lenses. AB - A challenge of the sutured posterior chamber intraocular lens (IOL) technique is to perform blind actions behind the iris. To avoid imprecise transscleral sutures and complications, we use an endoscopic procedure with 2 goals: to control the entry site of the needle penetration and of the haptic location. The endoscopic technique allows retroiris control during transscleral suturing and iridociliary IOL implantation. It is a safe, precise method that avoids the risks of blind procedures behind the iris. PMID- 10831893 TI - Retreatment after initial laser in situ keratomileusis. AB - PURPOSE: To report the results of laser in situ keratomileusis (LASIK) retreatment in patients with primary undercorrection and with postoperative regression and to assess the efficacy and safety of LASIK retreatment. SETTING: The Eye Institute of Utah, Salt Lake City, Utah, USA. METHODS: The prospective study included 157 eyes of 108 patients. Using the VISX Star laser, retreatments were performed for undercorrection in 43 eyes (27%) and for regression in 114 eyes (73%). The mean follow-up was 15 months (range 6 to 28 months) after the initial surgery and 10 months (range 3 to 25 months) after the repeat LASIK procedure. RESULTS: Overall, the mean spherical equivalent (SE) was -6.11 +/- 2.35 diopters (D) (range -1.87 to -15.00 D) before LASIK and -1.28 +/- 0.57 D (range -0.50 to -3.25 D) prior to retreatment. At the last visit, it was -0.23 +/ 0.41 D (range -2.55 to 1.13 D). One hundred fifty-three eyes (97.5%) were within +/-1.00 D of emmetropia and 128 (81.5%), within +/-0.50 D. The uncorrected visual acuity was 20/20 or better in 68.8% of eyes and 20/40 or better in 98.1%. In all eyes, the best corrected visual acuity (BCVA) improved or remained within 1 line of the pre-revision level. However, 2 eyes (1.3%) lost 2 or more lines of pre initial LASIK BCVA. Both eyes had a preoperative SE greater than -8.00 D; corneal irregularity was the cause of the decrease in vision. CONCLUSIONS: Several conclusions can be drawn from this study. First, LASIK retreatment is effective and safe for correcting undercorrection and regression after initial LASIK, and a good visual outcome is expected. Second, in most eyes, regression occurs within 6 months after the initial LASIK. However, regression can develop up to 2 years after LASIK. Third, multiple retreatments will be required in some patients. The outcome in these cases is promising. PMID- 10831894 TI - Excimer laser retreatment of residual myopia following photoastigmatic refractive keratectomy for compound myopic astigmatism. AB - PURPOSE: To prospectively evaluate the safety, efficacy, and visual performance of excimer laser enhancement following photoastigmatic refractive keratectomy (PARK) with the Summit Apex Plus. SETTING: Stanford University School of Medicine Eye Laser Clinic, Stanford, California, USA. METHODS: As part of a Food and Drug Administration clinical trial, 93 eyes of 56 patients with a mean spherical equivalent (SE) of -4.98 diopters (D) +/- 1.80 (SD) (range -1.75 to -8.50 D) had PARK for compound myopic astigmatism using the Summit Apex Plus excimer laser and a poly(methyl methacrylate) erodible mask. Seventeen eyes with a mean SE of -2.08 +/- 0.76 D required excimer laser refractive keratectomy for residual spherical myopia or compound myopic astigmatism. Patients were prospectively followed 1, 3, 6, 9, and 12+ months after the enhancement procedure. Primary outcome variables included uncorrected visual acuity (UCVA), refraction, vector analysis, best spectacle-corrected visual acuity (BSCVA) under standard ambient conditions (photopic, scotopic, and glare), corneal clarity, and contrast sensitivity function curve under photopic and scotopic conditions. RESULTS: At the last postoperative visit, the mean sphere had been corrected 82% to a residual of 0.29 +/- 1.23 D and mean SE had been corrected 65% to a residual of -0.74 +/- 1.27 D. Eighty-two percent of eyes were within +/-1.0 D of attempted correction. Eighty-eight percent had a UCVA of 20/40 or better. Vector analysis demonstrated a difference vector of within +/-1.0 D in 75% of eyes that had PARK retreatment. There was no significant loss in the contrast sensitivity curve. Late regression associated with corneal haze and loss of BSCVA occurred in 2 eyes (11.7%). CONCLUSIONS: Retreatment following PARK for compound myopic astigmatism results in effective reduction in residual spherical myopia and compound myopic astigmatism. An improvement in UCVA without loss of contrast sensitivity can be expected in most eyes. However, regression, corneal haze, and loss of BSCVA may occur. Further studies are indicated to predict risk factors for these complications. PMID- 10831895 TI - Intraocular lens power calculations in patients with extreme myopia. AB - PURPOSE: To determine the variables that might contribute to improved intraocular lens (IOL) power calculations preoperatively in cataract patients with extreme myopia. METHODS: This retrospective study included 50 patients with extreme myopia and axial lengths longer than 27.0 mm. All patients had clear corneal phacoemulsification by the same surgeon and implantation of the Domilens SiFlex 1 IOL (power range -6.0 to +5.0 diopters [D]). The performances of the SRK/T, Hoffer Q, Holladay 1, and Holladay 2 formulas in predicting an IOL power that would meet the target refraction of +/-1.00 D were compared. RESULTS: The formulas tended to suggest underpowered IOLs, more severe in eyes with axial lengths greater than 30.00 mm. These eyes accounted for most of the minus-power IOLs implanted. Back calculations of axial lengths in patients with minus-power IOLs showed that, on average, emmetropia could have been predicted by choosing shorter axial lengths (up to 2.72 mm shorter) than those used in the original IOL power calculations. Preoperative B-scan ultrasonography demonstrated the presence of a posterior pole staphyloma temporal to the optic nerve in several patients who required minus-power IOLs, which suggests that axial length measurement problems were a major source of IOL calculation errors in these patients. CONCLUSIONS: In eyes with axial lengths longer than or equal to 27.0 mm, current third- and fourth-generation lens calculation formulas have a tendency to over minus patients between -1.0 and -4.0 D. The formulas appear to perform better for plus-power IOL implantation than for minus-power IOL implantation. The use of B scan ultrasonography to locate posterior pole staphylomas may improve the accuracy of IOL calculations in eyes with extreme myopia. PMID- 10831896 TI - Automated keratoconus detection using the EyeSys videokeratoscope. AB - PURPOSE: To evaluate the effectiveness of indices derived from the EyeSys System 2000 in detecting keratoconic corneas. SETTING: Department of Ophthalmology, Hopital Saint Antoine, Paris VI University, Paris, France. METHODS: Topographies of 208 corneas were evaluated. The corneas were from 8 groups of patients classified by the following diagnoses: normal, regular astigmatism, cataract, radial keratotomy, photorefractive keratectomy, myopic keratomileusis, penetrating keratoplasty (PKP), and keratoconus. Nine statistical indices derived from EyeSys data, 2 Holladay Diagnosis Summary indices (coefficient of uniformity and coefficient of asphericity [Asph]), and our refractive power symmetry index were studied. A training set of 104 corneas was used to determine the most efficient threshold value of each index based on sensitivity and specificity curves. Decision trees combining 2 indices were generated. Sensitivity and specificity were calculated in a validation set composed of the remaining 104 corneas. RESULTS: Based on the results of the training set, the optimum indices were SDSD (standard deviation of the standard deviations of the radii of curvature of each ring) and Asph. In the validation set, the decision tree using these indices featured 88.5% sensitivity and 94.9% specificity; the 4 false positive cases were in corneas in the PKP group of patients. CONCLUSIONS: Clinically apparent keratoconus can be detected among normal corneas and irregular corneal shape patterns using the EyeSys System 2000 data and a decision tree combining 2 indices. PMID- 10831897 TI - Results of pediatric laser in situ keratomileusis. AB - PURPOSE: To evaluate the results of laser in situ keratomileusis (LASIK) for uniocular high myopia in pediatric eyes. SETTING: Dr. Agarwal's Eye Hospital, Chennai (Madras), India. METHODS: Sixteen eyes that were treated by LASIK for uniocular high myopia were retrospectively analyzed. The mean patient age was 8.4 years +/- 1.83 (SD) (range 5 to 11 years). Laser in situ keratomileusis was performed using the Technolas Keracor 217 excimer laser and the Automated Corneal Shaper microkeratome, which created a 160 microm corneal lamellar flap. Postoperatively, patients were reviewed at 1 day, 1 week, and 1, 6, and 12 months. The examination included uncorrected visual acuity (UCVA), best corrected visual acuity (BCVA), refraction, anterior segment evaluation, grading of haze based on a 5-point scale, intraocular pressure, corneal topography, and fundus evaluation. The Student t test was used for statistical analysis. RESULTS: The charts of all patients were analyzed at the 12 month visit. The mean preoperative spherical equivalent (SE) was -14.88 +/- 3.69 diopters (D) (range -9.00 to -23.00 D) and the mean postoperative SE, -1.44 +/- 1.14 D (range 0 to -2.50 D) (P < .05). The safety index was 1.01 (mean postoperative BCVA 0.54 and mean preoperative BCVA 0.53; P = .77). The efficacy index was 0.53 (mean postoperative UCVA 0.28 and mean preoperative BCVA 0.53). None of the eyes had an induced astigmatism of more than 0.5 D. Twelve eyes regained their BCVA, 2 lost 1 line of BCVA, and 2 gained 1 line. Three eyes had grade 2 haze. No retinal complications were observed. CONCLUSION: In this study, LASIK for uniocular high myopia in pediatric eyes provided encouraging results in the management of select cases of anisometropic amblyopia when other measures failed. A larger study with a longer follow-up is necessary to determine the long-term effects. PMID- 10831898 TI - Epithelial healing rates with topical ciprofloxacin, ofloxacin, and ofloxacin with artificial tears after photorefractive keratectomy. AB - PURPOSE: To determine whether corneal epithelial healing differs after the use of topical ciprofloxacin alone, topical ofloxacin alone, or topical ofloxacin with artificial tears in patients having photorefractive keratectomy (PRK). SETTING: Department of Ophthalmology and Visual Science, The University of Texas Health Science Center at Houston, Houston, Texas, USA. METHODS: Eighteen patients (6 women, 12 men) with moderate myopia (-1.50 to -6.00 diopters [D]) had standardized PRK. Patient age ranged from 25 to 62 years. The 28 eyes (16 right, 12 left) were randomized into 3 treatment groups: ofloxacin alone, n = 9 eyes; ciprofloxacin, n = 9 eyes; and ofloxacin with Refresh Plus, n = 10 eyes. The drugs were administered immediately after surgery and then every 6 hours. Video recordings of the corneal wounds stained with fluorescein were performed at 8:00 AM and 4:00 PM using a video slitlamp camera with a cobalt-blue light until the wound completely healed. The videotaped images were recorded and analyzed by a computer planimetry program. Wound areas were recorded and compared among the 3 drugs. The square-root transformation was applied to the wound area to obtain a constant healing rate. Statistical comparisons were analyzed using an analysis of variance test. RESULTS: Mean recovery time was 82.67 hours +/- 14.42 (SD) in the ofloxacin eyes, 120.89 +/- 34.05 hours in the ciprofloxacin eyes, and 76.80 +/- 19.30 hours in the ofloxacin with Refresh Plus eyes. Mean healing rate was 0.66 +/- 0.17 hours, 0.54 +/- 0.16 hours, and 0.67 +/- 0.15 hours, respectively. The healing rate was significantly higher in the ofloxacin with Refresh Plus eyes than in the ciprofloxacin eyes (P < .0001). There was no significant difference between the ofloxacin eyes and the ofloxacin with Refresh Plus eyes (P = .42). CONCLUSION: Ofloxacin with Refresh Plus and ofloxacin alone had a more positive effect on epithelial healing than ciprofloxacin. The ciprofloxacin eyes were significantly more prone to impaired or delayed wound healing and to the development of corneal haze. PMID- 10831899 TI - Nonpenetrating filtration surgery for glaucoma: control by surgery only. AB - PURPOSE: To report on intraocular pressure (IOP) control by nonpenetrating filtration surgery without medical treatment in eyes with primary open-angle glaucoma (POAG). SETTING: Oxford Eye Center and St. John Eye Hospital, Johannesburg, South Africa. METHODS: This retrospective study evaluated IOP control by nonpenetrating filtration surgery in 46 patients (86 eyes) with POAG who had surgery between February 1992 and February 1998 (mean follow-up 46 months); 48 eyes were previously treated, and 38 were newly diagnosed, untreated eyes. Under a trapezoidal scleral flap, a deep sclerectomy and a fenestration in Schlemm's canal were created without penetrating the anterior chamber. Postoperatively, when IOP rose above 20 mm Hg, instead of adding medical treatment, the filtration site was revised to re-establish filtration. RESULTS: The mean drop in IOP was 50%, from a mean of 30.4 mm Hg preoperatively to a mean of 15.35 mm Hg postoperatively. After a mean period of successful filtration of 29.9 months, the filtration site was revised in 48 eyes (56%) to maintain the IOP below 21 mm Hg without medication. The reoperation rate was 4.7 times higher in previously treated patients than in untreated patients. CONCLUSIONS: Nonpenetrating filtration surgery in eyes with POAG was more rewarding in untreated patients than in medically treated patients. To control IOP without medication, revision of the filtration site is feasible and needed in the medium and long term. PMID- 10831900 TI - T-cut in the bottom of the scleral pocket in combined cataract and glaucoma surgery. AB - PURPOSE: To determine whether radial transsection of the inner lamina of the phaco tunnel (T-cut) allows intraocular pressure (IOP) control in cases of co existing cataract and glaucoma and to evaluate the results of this glaucoma triple procedure. SETTING: Department of Ophthalmology, Hanusch Hospital, Vienna, Austria. METHODS: In a prospective study, a T-shaped incision in the tunnel floor was performed in 43 eyes (Group A); 48 eyes (Group B) had phacotrabeculectomy (phacoemulsification and trabeculectomy of a 3.0 x 2.0 mm tissue block). RESULTS: At the end of a minimum follow-up of 24 months in Group A (range 24 to 30 months) and of 28 months in Group B (range 28 to 44 months), there was no significant difference in the extent of IOP reduction between groups. Intraocular pressure was well controlled (< or =20 mm Hg) without antiglaucoma therapy in 27 eyes (62.8%) in Group A and 30 (62.5%) in Group B. Eleven eyes (25.6%) in Group A and 14 (29.2%) in Group B achieved an IOP of 20 mm Hg or less with antiglaucoma therapy. Five eyes (11.6%) in Group A and 4 (8.3%) in Group B required surgical revision. Complications included hypotony (Group A, 5 eyes; Group B, 3 eyes), hyphema (Group A, 8 eyes; Group B, 6 eyes), malignant glaucoma (Group B, 1 eye), in-the-bag hematoma (Group B, 1 eye), and fibrin exudation (Group B, 1 eye). CONCLUSION: Phacoemulsification with a T-cut in the tunnel floor was a safe and effective combined procedure and, in this regard, equivalent to phacotrabeculectomy. PMID- 10831901 TI - Transforming growth factor-beta isoform proteins in cell and matrix deposits on intraocular lenses. AB - PURPOSE: To determine whether the cells that adhere to poly(methyl methacrylate) (PMMA) posterior chamber intraocular lenses (PC IOLs) implanted in human eyes produce transforming growth factor-beta (TGF-beta) isoforms and whether the acellular proteinaceous deposits on these IOLs contain TGF-beta. SETTING: Department of Ophthalmology, Wakayama Medical College, Japan. METHODS: Thirty-two PMMA PC IOLs explanted from Japanese patients were immunostained for TGF-beta1, beta2, or -beta3, and observed under light microscopy. RESULTS: Cell deposits were observed on 12 IOLs and proteinaceous deposits on 16. Components of the cell deposits were mainly of macrophage origin. The cell and matrix deposits tested positive for each isoform of TGF-beta. CONCLUSION: The cells that adhered to implanted PMMA PC IOLs produced TGF-beta, and the extracellular matrix that accumulated on the surface of the IOLs contained TGF-beta. Transforming growth factor-beta from the cells on IOLs may influence the healing process of residual lens capsules after cataract surgery with IOL implantation. PMID- 10831902 TI - Double-masked prospective ocular safety study of a lens epithelial cell antibody to prevent posterior capsule opacification. AB - PURPOSE: To evaluate the intraocular safety of an immunoconjugate (MDX-RA) developed to prevent posterior capsule opacification (PCO) in human eyes. SETTING: St. Thomas's Hospital Eye Department, London, United Kingdom. METHODS: Twenty-six patients had phacoemulsification and implantation of an intraocular lens (IOL). All were randomly allocated at the end of surgery to receive a 0.1 mL placebo or 0.1 mL of the immunotoxin MDX-RA intracamerally. Two doses of the drug were tested: 8 patients with a low dose (50 units), 9 patients with a high dose (100 units), and 9 with placebo. Follow-up at days 1, 14, 30, 60, 90, and 180 consisted of visual acuity measured by the Early Treatment of Diabetic Retinopathy Study test, contrast sensitivity, aqueous flare, specular microscopy of the IOL's anterior surface, and corneal endothelial counts. The percentage area of PCO was measured from retroillumination images of the posterior capsule. RESULTS: There was no decrease in corneal endothelial cell count in toxin-treated patients. Early postoperative flare, anterior chamber cell count, and corneal pachymetry were higher in toxin-treated patients. The median percentage area of PCO at 1 year was 32.0 in the placebo group, 3.8 in the low-dose group, and 7.4 in the high-dose group (P = .06). CONCLUSION: This prospective, randomized, placebo-controlled trial confirmed that MDX-RA is safe for intraocular use and is of potential value for further clinical trials of the prevention of PCO. PMID- 10831903 TI - Posterior capsule opacification after phacoemulsification: foldable acrylic versus poly(methyl methacrylate) intraocular lenses. AB - PURPOSE: To study the effects of foldable acrylic and poly(methyl methacrylate) (PMMA) intraocular lens (IOL) implantation on posterior capsule opacification (PCO). SETTING: Department of Ophthalmology, Dokuz Eylul University School of Medicine, Izmir, Turkey. METHODS: This consecutive series comprised 157 eyes having phacoemulsification with implantation of a foldable acrylic IOL with rectangular optic edges and a 5.5 mm optic (AcrySof) (n = 80 eyes) or a PMMA IOL with rounded optic edges and 5.5 mm optic (n = 77). Evaluated were incidence, location, and degree of PCO. Mean postoperative follow-up was 17.8 months +/- 1.7 (SD) (range 16 to 22 months). RESULTS: Posterior capsule opacification occurred in 8.7% of eyes in the foldable acrylic IOL group and in 24.7% of eyes in the PMMA IOL group. The difference between the 2 groups was statistically significant (P < .01). Centrally located PCO was significantly lower in the acrylic group (P < .01). There were 3 eyes with severe PCO in the PMMA group and none in the acrylic group. Anterior capsule contraction and fibrosis were present in 4 eyes in the PMMA group but none in the acrylic group. Soemmering's ring cataract formation was detected in 3 eyes, all with the AcrySof IOL. CONCLUSION: In addition to its optic material and rectangular optic edges, the AcrySof IOL provides additional advantages in lowering the incidence of PCO compared with rounded-edge PMMA IOLs. PMID- 10831904 TI - Endothelial cell loss after phacoemulsification: relation to preoperative and intraoperative parameters. AB - PURPOSE: To evaluate the effect of the location of the corneoscleral tunnel incision as well as preoperative and intraoperative parameters on total and localized endothelial cell loss. SETTING: Department of Ophthalmology, Humboldt University of Berlin, Berlin, Germany. METHODS: Fifty consecutive patients scheduled for routine cataract surgery were selected prospectively for this clinical trial. Preoperatively, the axial length, anterior chamber depth, lens thickness, and astigmatism were measured. Phacoemulsification time and relative energy as well as total surgical time were recorded. With a specular microscope, endothelial cell counts were determined centrally, superiorly, and temporally preoperatively and 6 weeks and 6 and 12 months postoperatively. RESULTS: After 12 months, the mean overall central endothelial cell loss in all eyes was 8.5%. The mean endothelial cell loss was 11.9% in the lateral quadrant and 11.4% in the superior quadrant. There were no significant differences between superior and temporal surgical approaches in intraoperative parameters of phacoemulsification time, relative intensity of phacoemulsification, and surgical time. There were no significant differences in central endothelial cell loss or in the area localized in the quadrant of the positions of the corneal surgical site. The only risk factors found significant for higher endothelial cell loss were shorter axial length and longer phacoemulsification time. CONCLUSIONS: The location of corneoscleral incisions for phacoemulsification can be chosen according to the preoperative astigmatism without inducing additional adverse effects on the corneal endothelium. Shorter eyes have a significantly higher risk for greater endothelial cell loss. PMID- 10831905 TI - Does preservative-free lignocaine 1% for hydrodissection reduce pain during phacoemulsification? AB - PURPOSE: To compare preservative-free 1% lignocaine with balanced salt solution (BSS) in alleviating pain during hydrodissection in phacoemulsification cataract surgery. SETTING: West Norwich Hospital, Norfolk, United Kingdom. METHODS: This prospective double-masked trial comprised 68 patients having day-case phacoemulsification cataract surgery. Patients were randomly divided into 2 groups, receiving either BSS or lignocaine 1% solution for hydrodissection during routine uneventful phacoemulsification using topical anesthesia. The level of intraoperative pain was scored on a scale of 0 (no pain) to 10 (severe pain), and the scores between the 2 groups were compared. RESULTS: Of the 68 patients, 33 (49%) received BSS and 35 (51%), lignocaine 1% solution. A pain score greater than 2 was considered clinically significant; 28 patients (85%) in the BSS group and 25 (71%) in the lignocaine 1% group scored 2 or less. The chi-square and Mann Whitney tests found no significant difference between the BSS and lignocaine 1% groups (P = .30 and P = .432, respectively). CONCLUSION: There was no significant difference in the pain scores in patients who received BSS or lignocaine 1% solution. Thus, we conclude that hydrodissecting with lignocaine 1% solution does not provide added pain relief during phacoemulsification. PMID- 10831906 TI - Effect of corneal thickness on the accuracy of intraocular pressure measurement in rabbits after excimer laser photoablation. AB - PURPOSE: To determine whether intraocular pressure (IOP) measurements using pneumotonometry and Tono-Pen tonometry are accurate after excimer laser photoablation of the central cornea in rabbits. SETTING: Department of Ophthalmology, Scott & White Hospital, Texas A & M University College of Medicine, Temple, Texas, USA. METHODS: Ten rabbits had excimer laser phototherapeutic keratectomy (PTK) in the right eye to thin the central corneal thickness (CCT) by approximately 20%. The left eye served as the control. The diameter of each PTK was 6.0 mm. Four weeks later, the rabbits were examined under general anesthesia. The eyes were cannulated, and IOP was maintained at 10, 20, and 30 mm Hg and measured using an IOP transducer, Tono-Pen, and pneumotonometer at each pressure level. The readings were compared. RESULTS: One rabbit was excluded from the study because of recurrent erosions and corneal edema. There was no statistically significant difference in measurement error between the operated eye and control eye for the Tono-Pen (P = .12) or pneumotonometer (P = .35). There were significant differences in measurement error as pressure increased for the Tono-Pen (P < .01) and pneumotonometer (P < .01). The increase in measurement error with the increase in pressure was similar in the 2 eyes: Tono-Pen (P = .96) and pneumotonometer (P = .55). A paired t test showed the observed CCT was comparable (P = .79) in the pre-laser right eyes (mean 400.56 microm +/- 11.29 [SD]) and control eyes (mean 401.78 +/- 19.78 microm). CONCLUSION: Photoablation of approximately 20% of the CCT in rabbits by excimer laser PTK did not significantly alter the accuracy of IOP measurements by the Tono-Pen or pneumotonometer. PMID- 10831907 TI - Influence of temperature and time on thermally induced forces in corneal collagen and the effect on laser thermokeratoplasty. AB - PURPOSE: To investigate thermomechanical aspects of corneal collagen denaturation as a function of temperature and time and the effect of the induced forces on refractive changes with laser thermokeratoplasty (LTK). SETTING: Medical Laser Center Lubeck, Lubeck, Germany. METHODS: In a material-test setup, porcine corneal strips were denatured in paraffin oil at various constant temperatures for 10 and 500 seconds, and the temporal course of the contractive forces was studied under isometric conditions. Typical LTK lesions were performed in porcine eyes in vitro with a continuous-wave infrared laser diode at a wavelength of 1.87 microm for 10 and 60 seconds. The laser power was chosen to achieve comparable denatured volumes at both irradiation times. The refractive changes were measured and analyzed by histologic evaluations and temperature calculations. RESULTS: The time course of the induced forces was characterized by a maximal force, which increased almost linearly with temperature, and a residual lower force. After 500 seconds of heating, the highest force was achieved with a temperature of 75 degrees C. With a limited heating period of only 10 seconds, the forces steadily increased with temperature over the entire observation period. Laser thermokeratoplasty produced less refractive change after 10 seconds of irradiation than after 60 seconds, although the laser power was 25% higher in the short heating period. Polarization light microscopy of LTK lesions revealed different stages of thermal damage. CONCLUSION: The course of the contractive forces during and after heating is a complicated function of the spatial time/temperature profile. Laser thermokeratoplasty lesions produced with 2 irradiation times showed different stages of denaturation and induced refractive change. PMID- 10831908 TI - Glaucoma laser treatment parameters and practices of ASCRS members--1999 survey. American Society of Cataract and Refractive Surgery. AB - A survey of glaucoma laser parameters and practices was mailed to all United States members of the American Society of Cataract and Refractive Surgery (ASCRS) in March 1999. Approximately 19% (947) of the 5100 surveys were returned. The survey assessed parameters used for laser trabeculoplasty, laser iridotomy, and gonioplasty. Respondents also answered questions about the application of these laser therapies in a wide variety of clinical situations. The results were cross tabulated by age, number of laser trabeculoplasties performed per month, fellowship training in glaucoma, and geographic location. PMID- 10831909 TI - Complications of foldable intraocular lenses requiring explanation or secondary intervention--1998 survey. AB - A survey of the complications associated with foldable intraocular lenses (IOLs) that required explantation or secondary intervention was sent to domestic members of the American Society of Cataract and Refractive Surgery in 1998. Preoperative data as well as patient signs and symptoms were obtained and postoperative results and outcomes assessed. The types of foldable IOLs explanted were tabulated. The percentage of each type removed and the common reasons for removal were as follows: 38%--3-piece monofocal silicone IOLs for incorrect lens power followed by dislocation/decentration; 31%--3-piece multifocal silicone IOLs for glare/optical aberrations; 16%--3-piece acrylic IOLs for glare/optical aberrations followed by incorrect lens power and dislocation; 15%--1-piece or plate-haptic silicone IOLs for dislocation/decentration followed by incorrect lens power. Visual outcomes were uniformly good in the 4 groups. Meticulous surgical technique, IOL insertion, and IOL power measurements, along with proper patient selection, are important factors in avoiding complications with foldable IOLs. PMID- 10831910 TI - Exchange of a posterior chamber phakic intraocular lens in a highly myopic eye. AB - A 38-year-old woman had posterior chamber phakic intraocular lens (IOL) implantation as a secondary refractive procedure to correct residual refraction (20/50 with -16.50 -1.50 x 80) in May 1998, 3 years after intrastromal corneal ring segment surgery for high myopia (-30.00 diopters). Ultrasound biomicroscopy revealed an oversized lens, leading to malpositioning. Moreover, the patient remained undercorrected (20/40 with -5.25 -0.75 x 120). Ten months later, the phakic IOL was uneventfully exchanged for a shorter one with the correct dioptric power. It was well placed in the posterior chamber. The patient's visual acuity was 20/30 with -2.25 -0.75 x 145, very close to the refraction in the fellow aphakic eye (20/30 with -2.50 -0.75 x 75). Patient satisfaction with the final visual outcome was high. Accurate ciliary sulcus measurement is critical for proper phakic IOL sizing. PMID- 10831911 TI - Postoperative refractive error resulting from incorrectly labeled intraocular lens power. AB - Postoperative refractive errors after intraocular lens (IOL) implantation can be caused by different reasons. The most likely is incorrect IOL calculation resulting from incorrect measurements of the eye. However, other explanations must also be taken into account. The surgeon in the operating theater should make sure that the correct IOL was chosen. The IOL package should also contain the correct IOL cartridge. When unsealed IOL packages are used, an IOL cartridge from 1 package can be mistakenly placed in another package. Finally, incorrect IOL labeling by the manufacturer can occur. In this case, the optical power of an explanted IOL was not identical to the IOL power printed on the company's label. Even with the highest quality control throughout the IOL manufacturing process, the surgeon should keep in mind the possibility of a mislabeled IOL. PMID- 10831912 TI - Spontaneous regression of Elschnig pearl posterior capsule opacification. AB - After extracapsular cataract extraction with in-the-bag intraocular lens implantation, a 72-year-old woman had reduced visual acuity from posterior capsule opacification (PCO) resulting from Elschnig pearl proliferation. No capsulotomy was performed, and the PCO decreased spontaneously over time, improving visual acuity and leaving a clear capsule. PMID- 10831913 TI - Clear lens extraction and intraocular lens implantation in Marfan's syndrome. AB - Intracapsular clear crystalline lens extraction and intraocular lens (IOL) implantation were performed in 4 highly myopic eyes of 2 patients with Marfan's syndrome. One eye of each patient received an anterior chamber IOL and the other, a scleral-fixated posterior chamber IOL. The preoperative spherical equivalent ranged between -14.50 and -28.00 diopters (D) and axial length range, between 25.32 and 36.02 mm. The SRK II formula was used. Mean uncorrected visual acuity improved from counting fingers to 20/80. Postoperative spherical equivalent correction ranged from -0.75 to +2.75 D. One eye had vitreous loss that was managed by anterior vitrectomy. Modern surgery for cataract and management of its complications suggest that clear crystalline lens extraction and IOL implantation can be attempted in selected cases with Marfan's syndrome. PMID- 10831914 TI - Scleral ectasia as a complication of deep sclerectomy. AB - Nonpenetrating deep sclerectomy is a filtration surgical technique to treat glaucoma. A 12-year-old girl presented with chronic arthritis complicated with glaucoma secondary to a chronic uveitis. A sclerectomy without a collagen implant was performed for uncontrollable glaucoma with deterioration in visual function. Three weeks later, the patient had a rise in intraocular pressure and a scleral ectasia on the sclerectomy. The eye showed an area of scleral ectasia in the bleb as well as iris adhesion. Partial resection of the bleb after iris detachment led to poor anatomic and IOP results. The indications for deep sclerectomy must be carefully considered, especially in patients at a young age with this type of glaucoma. PMID- 10831915 TI - Editorial. PMID- 10831916 TI - Pharmacological therapies to minimise platelet transfusion. PMID- 10831917 TI - Preliminary experience with plasma exchange in patients with ulcerative colitis. AB - We decided to test the effect of plasma exchange (PE) in selected patients with inflammatory bowel disease in which conventional medical treatment proved insufficient. Twenty-six patients with ulcerative colitis (UC) were treated with PE, six patients with fulminant colitis and 20 with long lasting severe colitis. After very promising results initially, 17 patients with long lasting severe colitis were selected in a careful prospective study. Twelve patients completed the treatment protocol. Two litres of plasma were exchanged every second or third day, 5-6 times in two weeks. The replacement fluid was fresh frozen plasma. The activity indices and histological evaluation were used as criteriae for treatment response. A reduction of 35% in the activity index is considered a significant improvement. In all but three patients (out of 12), the activity indices were reduced two weeks after end of treatment. Six patients had a reduction of more than 35%, three patients had a reduction between 22-28% and three had a reduction less than 10%. None became worse. In this carefully studied group, the immediate beneficial effects of PE was only demonstrable by the activity indices and not evident by blind evaluation of biopsies from the mucosa. In two of the six patients with fulminant colitis, PE was followed by an immediate dramatic clinical improvement. In the follow up period (2-14 years) 14 of the 26 patients reported marked and long lasting improvement of inflammatory bowel disease. We conclude that PE might be beneficial in subsets of patients with UC. PMID- 10831918 TI - PCR testing for HCV in anti-HCV negative blood donors involved in the so called HCV +ve post-transfusion hepatitis. AB - Although it is infrequent, post-transfusion HCV infection may occur if the donors blood is collected in the window period between exposure and anti-HCV detectability by ELISA testing. STUDY DESIGN: In these last years, despite of routine application of anti-HCV testing, our blood transfusion center has been involved in 53 cases of alleged post-transfusion HCV hepatitis and look-back programs were set up with the goal of finding out the donors possibly involved in viral transmission. Most of these patients were hematological cases with multiple transfusions given because of aplastic anemia (3 cases), leukemia with or without bone marrow transplantation (5/4 cases) but necessitating long-term platelet support, leukemia and solid cancer patients undergoing autologous PBSC transplantation (3/4 cases) and TTP (2 cases). Only 32 patients were of the simple medical or surgical type, 9 transfused because of cardiac or vascular surgery, 8 because of spine surgery, 5 for different diseases and 5 for different types of cancer surgery. Donor's infectivity was determined by ELISA anti-HCV testing, by recombinant immunoblotting assay, and by nucleic acid testing. RESULTS AND CONCLUSION: No donors out of 267 traced of a total of 359 involved was found with anti-HCV seroconversion, or positive on PCR testing. This suggests that the responsibility for HCV transmission can only hypothetically be related to blood or blood components and that other transmission routes should be found out. PMID- 10831919 TI - The role of in process qualification in quality improvement of the haemonetics MCS plus leucodepleted platelet concentrate. AB - With the implementation of universal leucodepletion in UK all leucodepletion processes have gone through a standard process qualification and quality improvement. The Haemonetics MCS system is a well established automated platelet collection system for the production of double dose leucoreduced platelet concentrate (WBC approximately 70x10(6)/dose). Recently an automated post collection filtration harness system has been introduced (MCS plus LDP) in which platelets are filtered, using an in-line PALL polyester filter (LRFH6 PALL) to reduce the WBC level to below 5x10(6) WBC/dose. This system passed our Phase I evaluation process based on 20-40 runs. However, some changes in the final volume of the products were needed to conform to national guidelines. Large scale trials using the new volume adjusted protocol revealed occasional failure in the leucocyte content. Therefore, 100% testing had to be implemented on all products. A national evaluation was carried out to determine whether changing the filter to a more efficacious one, the LRFXL (PALL) or slowing the filtration flow rate can influence the overall outcome. To reduce donor variability, known donor population were used with identical apheresis conditions. A more consistent and systematic drop in leucocyte content was observed by reducing the flow rate whereas a similar failure (i.e. 1-3%) rate was found both in controls and LRFXL when using the standard head pressure, which is recommended by the manufacturer. A similar failure rate was found using three different low leucocyte counting technologies (Nageotte, flow cytometry and Imagn 2000). It is recommended that a process qualification/validation program should be implemented when even a small modification in the collection system is introduced. PMID- 10831920 TI - The supply of blood products in 10 different systems or countries. AB - Countries vary greatly in their ability to produce their own blood products including albumin and IVIgG. Part of this variability depends on the supply of plasma within the country. As has been seen most recently in the UK, the quality of the plasma and its acceptability for plasma fractionation must also be considered. Therefore concerns regarding the quality of the plasma have been added to those regarding the quantity.Only a few countries are nationally self sufficient in plasma. This has a marked effect on blood product availability and therefore the ability to treat patients. Unlike most pharmaceuticals, the plasma fractionation industry must rely, for its raw products, on plasma obtained from blood donors. As such this puts it in a potentially compromised situation since neither the supply nor the quality of the raw material can be assured and both of those will vary with time. This paper reviews the processes through which blood products are made available in 10 different systems including: Canada, England, France, Italy, Norway Scotland, Sweden, Switzerland, South Africa and USA. A series of specific questions were posed and the responses received from the various coauthors and other respondents provide comparative data on blood product availability in different areas of the world. PMID- 10831921 TI - Clinical application of therapeutic erythrocytapheresis (TEA). AB - Therapeutic erythrocytapheresis (TEA) has been used in different diseases such as polycythemia vera (PV), secondary erythrocytosis or hemochromatosis as a process of the less cumbersome but more expensive phlebotomy. TEA is preferred in emergency conditions such as thrombocytosis or in conditions such as porphyria cutanea tarda (PCT) or erythropoietic porphyria when plasma exchange (PEX) is often combined with TEA to reduce extracellular levels of uroporphyrin which contribute to plasma hyperviscosity. TEA is often combined with drug therapy that varies from etoposide in PV to EPO and desferoxamine which are used to mobilize and reduce iron stores in hemochromatosis. Benefits from this combination may be more long lasting than expected. Nonetheless for TEA, there is no standard protocol and, clinical experience with this therapy remains highly anecdotal. Therapeutic red cell-exchange (TREX) has been used with much interest over the years, starting with the management of hemolytic disease of the newborn and later used to correct severe anemia in thalassemia patients thereby preventing iron overload. It has also been used for the management of complications of sickle cell disease such as priapism, chest syndrome, stroke, retinal, bone, splenic and hepatic infarction or in preparation for surgery by reducing HbS to less than 30%. Automated apheresis has also favored the use of TREX in conditions such as paroxysmal nocturnal hemoglobinuria and aniline poisoning, arsenic poisoning, Na chlorate intoxications and CO intoxications, hemoglobinopathies, autoimmune hemolytic anemia, reactions due to ABO incompatibility, in preparation for ABO incompatible bone marrow transplantation or for preventing anti-D immunization after the transfusion of D(+) cells to D(-) recipients. Another field of application has been in the emergency management of intraerythrocytic parasite infections such as malaria and babesiosis. Application of TREX may be wide but its real use remains limited. In our personal experience, in 16 years, only 167 TREX procedures have been carried out in a total of 13,747 therapeutic procedures. This represents only 1.21% of the total. PMID- 10831922 TI - National survey of hemapheresis practice in Turkey (1998). AB - The Turkish Apheresis Group has maintained a national registry for apheresis activities since 1997. The hemapheresis practice of Turkey in 1998 is summarized in brief detail in this article. A total of 30, 136 apheresis procedures were performed at 31 different apheresis centers. At 10 centers, 145 peripheral blood stem cell (PBSC) apheresis were performed on 82 patients in allogeneic setting and at 17 centers, 981 PBSC apheresis were performed on 271 patients in autologous setting. Frequently observed adverse effects during PBSC apheresis were mild tremor and chills, paresthesia and nausea in 15% of the patients and donors. Vascular access complications, particularly observed in autologous setting due to central venous catheters were encountered in 10% of the procedures. Eight hundred and sixty-nine therapeutic plasma exchange procedures were performed at 21 centers on 172 patients, most commonly for neurological disorders and thrombotic thrombocytopenic purpura (TTP)/hemolytic uremic syndrome (HUS). Therapeutic cytapheresis procedures like leukapheresis, plateletapheresis and erythrocyte apheresis were performed especially for cytoreduction in myeloproliferative disorders. A total of 204 cytapheresis procedures (66% leukapheresis, 33% plateletapheresis and 1% erythrocytapheresis) were performed on 134 patients in 15 centers. Donor plateletapheresis was the most used apheresis procedure, reaching a total of 28.016 in 1998. Many university hospitals and a few state hospitals are performing above-mentioned apheresis procedures with great success and acceptable side effects. According to these data we are planning prospective trials and will establish National Standards of Practice. PMID- 10831923 TI - UK strategy for process qualification/validation: from concept to practical implementation. AB - In this communication, after a brief review of current requirements of various stages of leucodepletion qualification/validation processes, we provide some practical examples for the usefulness of long term validation programme for process improvement. It is hoped that the experience gained from this process qualification/validation enhances the awareness to key variables that can influence the overall outcome of filtration programme. PMID- 10831924 TI - Are biochemical markers for bone turnover clinically useful for monitoring therapy in individual osteoporotic patients? PMID- 10831925 TI - Monitoring individual response to hormone replacement therapy with bone markers. AB - Hormone replacement therapy (HRT) induces a rapid decrease in biochemical markers of bone turnover that correlate with a subsequent increase in bone mineral density (BMD). To determine the utility of bone markers in the management of postmenopausal women receiving HRT, we analyzed the relationship between changes in four markers (serum osteocalcin and bone alkaline phosphatase [BAP], serum and urinary C-telopeptide of type I collagen [CTX]) and changes in spine BMD in 569 women treated for 2 years with different doses of a matrix transdermal 17beta estradiol patch in two placebo-controlled trials. Using a logistic regression model, we found that both the percent change from baseline and the actual value of resorption markers at 3 and 6 months of treatment were predictive of BMD response at 2 years. Comparable results were obtained with formation markers at 6 months only. We determined the sensitivity, probably of positive BMD response, and corresponding cutoff value of markers at 3 and 6 months with a specificity set at a level of 0.90, so that <10% of women classified with markers as responders, i.e., as having a subsequent increase in BMD at 2 years >/=2.26%, would be false positive. All markers provided a high probability of positive BMD response ranging from 0.82 to 0.91, with a sensitivity higher for resorption than for formation markers, and sometimes improved in a model combining the percent change and the actual value of marker under HRT. For example, a decrease in serum CTX >/= 33% at 3 months of HRT provided a 68% sensitivity and 87% probability of positive BMD response at 2 years for a 90% specificity. At 6 months, a decrease in urinary CTX >/= 53% provided a 68% sensitivity and 91% probability of a positive BMD response for a 90% specificity. Half of false-negative cases at 3 months will be correctly identified by a subsequent urinary CTX measurement at 6 months. We conclude that the short-term change in bone markers reflects long-term changes of BMD in postmenopausal women treated with HRT. Our data suggest that bone turnover markers can be used to monitor the BMD response to HRT at the individual level. Whether such monitoring could improve long-term compliance to HRT should be tested prospectively. PMID- 10831926 TI - Early response in biochemical markers predicts long-term response in bone mass during hormone replacement therapy in early postmenopausal women. AB - Based on data from 153 early postmenopausal women who completed a double-blind, randomized 3 year study of graded hormone replacement therapy (HRT) doses or placebo, we investigated the value of bone markers to predict prevention of bone loss. Absolute values of serum and urinary CrossLaps (S-CTX and U-CTX) after 2 weeks of treatment were significantly correlated to 3 year bone mass response (r = -0. 28/-0.35; p < 0.001). These associations were fully expressed at 6 months (r = -0.61/-0.64; p < 0.001). Receiver operating characteristic analyses revealed that the predictive capacity of one measurement of a resorption marker after 6 months' treatment performed similarly as assessment of hip bone mass over 3 years in predicting preservation of spinal bone mass over 3 years. Comparable results were obtained using percent change from baseline in resorption markers at both 6 and 12 months, whereas for formation markers percent change was superior to absolute value at 6 months but not at 12 months. Values of accuracy for S-CTX for a cutoff of 1881 pmol/L at 6 months were 85.2% (sensitivity), 74.3% (specificity), 90.5% (positive predictive value), and 63.4% (negative predictive value); U-CTX performed similarly, whereas the values for the formation markers were slightly lower. A cutoff for S-CTX of 1245 pmol/L eliminated false-positive individuals (those who had a decrease below the cutoff but lost bone). In the false-negative group, which was composed of individuals whose S-CTX did not decrease below the cutoff but had preserved bone mass, S-CTX was significantly associated with spinal bone mass response (r = -0. 41; p < 0.01), indicating these women had been treated with a dose that was not at its optimum for their individual bone turnover. For this cutoff, the values were 49.5% (sensitivity), 97.1% (specificity), 98% (positive predictive value), and 40% (negative predictive value). In conclusion, early bone marker measurements predict long term preservation of bone mass during HRT. Resorption markers seem superior to formation markers, which reflects that the primary effect of HRT is on bone resorption. A strategy with two cutoff levels may optimize the use of bone markers to predict bone mass response. Whether resorption markers can be used to guide individualized treatment remains to be investigated. PMID- 10831927 TI - Components of biological variation of biochemical markers of bone turnover in Paget's bone disease. AB - The aims of this study were to evaluate the components of biological variation of the new markers of bone turnover in patients with Paget's bone disease and to compare the results with data obtained in healthy subjects. Fifteen patients with Paget's disease in a stable period of the disease and 12 healthy premenopausal women were included for a 1 year follow-up study. Within- and between-subject biological variation, indices of individuality, and critical differences were evaluated for the following biochemical markers: in serum, total (tAP), and bone (bAP) alkaline phosphatases, procollagen type I N-terminal propeptide (PINP) and beta-carboxyterminal telopeptide of type I collagen (sCTx); in urine, hydroxyproline (Hyp), and amino (NTx) and beta-carboxyterminal (CTx) telopeptides of collagen type I. Serum markers of bone turnover showed lower biological variability than urinary markers. Within-subject biological variation was higher in pagetic patients than in healthy subjects for all serum markers. In both groups, bAP presented the lowest within-subject biological variation. In pagetic patients, all markers presented indices of individuality of <0.6, indicating their usefulness for patient monitoring. Critical differences were lower for serum markers than for urinary markers. Among pagetic patients, serum bAP and PINP showed the lowest critical differences with values close to 30%, whereas urinary CTx presented the highest critical differences (near 70%). Conversely, in healthy subjects, tAP was the marker with the lowest critical differences, being two-fold higher in pagetic patients. This study confirms the lower sensitivity of urinary markers to detect significant changes and indicates that data obtained on biological variations from healthy populations cannot always be extrapolated to pathological conditions. In addition, serum bAP and PINP seem to be the markers that best reflect a significant change in activity of Paget's disease. PMID- 10831928 TI - Familial Paget's disease of bone: patterns of inheritance and frequency of linkage to chromosome 18q. AB - Paget's disease of bone is a common disorder characterized by focal abnormalities of bone turnover which are associated with bone pain bone deformity and an increased risk of pathological fracture. Genetic factors play an important role in the pathogenesis of Paget's disease, and recent genetic linkage studies have shown that in some families the disease is linked to a candidate locus on chromosome 18q21-22, which also harbors the gene for the related inherited condition, familial expansile osteolysis. In this study we characterized the patterns of inheritance in a series of 269 individuals from a further 50 kindreds with familial Paget's disease and sought to determine how frequently the disease was linked to chromosome 18q. Segregation analysis showed that 54% of individuals had developed Paget's disease by the age of 55, with an equal distribution in men in women, consistent with an autosomal dominant mode of inheritance with high penetrance. In families where parental data were available, there was no difference in the frequency of disease transmission between paternal or maternal sources. Linkage studies with nine polymorphic markers spread across the candidate region did not support linkage to 18q under models of homogeneity or heterogeneity. Indeed, the summated multipoint lodscores were consistently below 2.0 across the region, providing strong evidence against linkage. These studies confirm the presence of genetic heterogeneity in familial Paget's disease but show that linkage of the disease to the previously identified candidate locus on chromosome 18q21-22 is relatively uncommon. PMID- 10831929 TI - Static and dynamic bone histomorphometry in children with osteogenesis imperfecta. AB - Osteogenesis imperfecta (OI) is a genetic disorder characterized by increased bone fragility and low bone mass. Four clinical types are commonly distinguished. Schematically, type I is the mildest phenotype, type II is usually lethal, type III is the most severe form compatible with postnatal survival, and type IV is moderately severe. Although mutations affecting collagen type I are responsible for the disease in most patients, the mechanisms by which the genetic defects cause abnormal bone development have not been well characterized. Therefore, we evaluated quantitative static and dynamic histomorphometric parameters in tetracycline-labeled iliac bone biopsies from 70 children, aged 1.5 to 13.5 years, with OI types I (n = 32), III (n = 11), and IV (n = 27). Results were compared with those of 27 age-matched controls without metabolic bone disease. Biopsy core width, cortical width, and cancellous bone volume were clearly decreased in all OI types. Decreased cancellous bone volume was due to a 41%-57% reduction in trabecular number and a 15%-27% lower trabecular thickness. Regression analyses revealed that trabecular number did not vary with age in either controls or OI patients, indicating that no trabecular loss occurred. The annual increase in trabecular thickness was 5.8 microm in controls and 3.6 microm in type I OI, whereas no trabecular thickening was evident in type III and IV OI. Wall thickness, which reflects the amount of bone formed during a remodeling cycle, was decreased by 14% in a subgroup of 17 type I OI patients, but was not determined in the other OI types. The remodeling balance was less positive in type I OI than in controls, and probably close to zero in types III and IV. Surface-based parameters of bone remodeling were increased in all OI types, indicating increased recruitment of remodeling units. No defect in matrix mineralization was found. In conclusion, there was evidence of defects in all three mechanisms, which normally lead to an increase in bone mass during childhood; that is, modeling of external bone size and shape, production of secondary trabeculae by endochondral ossification, and thickening of secondary trabeculae by remodeling. Thus, OI might be regarded as a disease in which a single genetic defect in the osteoblast interferes with multiple mechanisms that normally ensure adaptation of the skeleton to the increasing mechanical needs during growth. PMID- 10831930 TI - Determinants of bone density and prevalence of osteopenia among female runners in their second to seventh decades of age. AB - This is a cross-sectional study of spine and hip bone density (BMD) in 124 female athletes, aged 16-68 years, who trained for at least 3 hs/week. The aim was to document the effects of competitive running on BMD in women over a broad age range. Thirty-three subjects, aged <35 years, were currently oligo- or amenorrheic and, of the 50 who were >40 years, and who were now menstruating normally, 13 had previously been oligo- or amenorrheic. Fifty-two women <50 years of age had never had disturbed menses. Twenty-four older women were postmenopausal. Women who had never had menstrual disturbance had significantly increased bone density at the lumbar spine, femoral neck, and femoral trochanter, as compared with young normal European reference data (range from +0.4 population SD or T-score units to +1. 2 units according to measurement site and age group). In contrast, young amenorrheic or oligomenorrheic runners had reduced bone density, particularly at the spine (mean T score < -1.1), whereas older runners who previously had disturbed menses, but were now menstruating normally, had bone densities that were similar to sedentary young controls. Postmenopausal runners had bone density values that differed little from sedentary postmenopausal controls matched for time since menopause, after adjusting for the runners' lower body weight. Bone density outcomes were related to candidate explanatory variables. After taking into account the other variables, age, per se, influenced only the femoral neck and Ward's area. Years since last exposure to estrogen (at premenopausal levels) was an important determinant of bone loss at both hip and spine. Body weight had a beneficial influence on the femoral neck region, whereas (in contrast) height had a positive influence on the lumbar spine. Months of breastfeeding (totaled for all children) had a modest, positive influence, which was larger in the femoral measurement sites. There was no evidence of an effect of calcium intake or percent body fat on BMD at any site independent of these other effects. It is concluded that, with the consistent presence of normal premenopausal estrogen levels, running at least 3 hs/week substantially improves bone density, particularly at the proximal femur. This beneficial effect is reversed in the absence of the consistent past and current presence of normal menstrual function. There was no clear benefit of running seen on BMD in postmenopausal women, but premenopausal veteran athletes who started running after the age of 30 years were not disadvantaged compared with early starters. PMID- 10831931 TI - Evidence of hydroxyl-ion deficiency in bone apatites: an inelastic neutron scattering study. AB - The novelty of very large neutron-scattering intensity from the nuclear-spin incoherence in hydrogen has permitted the determination of atomic motion of hydrogen in synthetic hydroxyapatite and in deproteinated isolated apatite crystals of bovine and rat bone without the interference of vibrational modes from other structural units. From an inelastic neutron-scattering experiment, we found no sharp excitations characteristic of the vibrational mode and stretch vibrations of OH ions around 80 and 450 meV (645 and 3630 cm(-1)), respectively, in the isolated, deproteinated crystals of bone apatites; such salient features were clearly seen in micron- and nanometer-size crystals of pure hydroxyapatite powders. Thus, the data provide additional definitive evidence for the lack of OH(-) ions in the crystals of bone apatite. Weak features at 160-180 and 376 meV, which are clearly observed in the apatite crystals of rat bone and possibly in adult mature bovine bone, but to a much lesser degree, but not in the synthetic hydroxyapatite, are assigned to the deformation and stretch modes of OH ions belonging to HPO(4)-like species. PMID- 10831932 TI - Heterogeneity of bone lamellar-level elastic moduli. AB - Advances in our ability to assess fracture risk, predict implant success, and evaluate new therapies for bone metabolic and remodeling disorders depend on our understanding of anatomically specific measures of local tissue mechanical properties near and surrounding bone cells. Using nanoindentation, we have quantified elastic modulus and hardness of human lamellar bone tissue as a function of tissue microstructures and anatomic location. Cortical and trabecular bone specimens were obtained from the femoral neck and diaphysis, distal radius, and fifth lumbar vertebra of ten male subjects (aged 40-85 years). Tissue was tested under moist conditions at room temperature to a maximum depth of 500 nm with a loading rate of 10 nm/sec. Diaphyseal tissue was found to have greater elastic modulus and hardness than metaphyseal tissues for all microstructures, whereas interstitial elastic modulus and hardness did not differ significantly between metaphyses. Trabecular bone varied across locations, with the femoral neck having greater lamellar-level elastic modulus and hardness than the distal radius, which had greater properties than the fifth lumbar vertebra. Osteonal, interstitial, and primary lamellar tissues of compact bone had greater elastic moduli and hardnesses than trabecular bone when comparing within an anatomic location. Only femoral neck interstitial tissue had a greater elastic modulus than its osteonal counterpart, which suggests that microstructural distinctions can vary with anatomical location and may reflect differences in the average tissue age of cortical bone or mineral and collagen organization. PMID- 10831933 TI - Temporal and spatial expression of bone morphogenetic protein-2, -4, and -7 during distraction osteogenesis in rabbits. AB - The Ilizarov method of limb lengthening makes use of the fact that osteogenesis is induced at an osteotomy site when distraction is applied. It is unknown at present how the mechanical forces created by distraction are translated into biological signals. Because bone morphogenetic proteins (BMPs) are potent inducers of osteogenesis in many experimental systems, they are obvious candidates for playing a role in this process. In this study, we investigated the temporal and spatial expression of BMP-2, -4, and -7 proteins during distraction osteogenesis using immunohistochemistry. An osteotomy was performed on the right tibiae of white New Zealand rabbits. After a delay of 7 days, distraction was started at a rate of 0.25 mm/12 h for 3 weeks, followed by a 3 week consolidation phase. Each week after osteotomy one rabbit was killed for immunohistochemical studies. Staining for BMP-2, -4, and -7 was evident before distraction was applied and was mainly localized to mesenchymal cells and osteoblastic cells in the periosteal region. After distraction was started, the typical fibrous interzone developed between the osteotomy fragments, where both intramembranous and endochondral ossification were noted. In this area, cells resembling fibroblasts and chondrocytes, but not mature osteoblasts, showed intense staining for all three BMPs. This high level of expression was maintained during the entire distraction phase and then gradually disappeared during the consolidation phase. These results are compatible with the hypothesis that BMPs play an important role in the signaling pathways that link the mechanical forces created by distraction to biological responses. PMID- 10831934 TI - Effects of locally applied transforming growth factor-beta1 on distraction osteogenesis in a rabbit limb-lengthening model. AB - In this study we tested the effect of locally applied transforming growth factor beta1 (TGF-beta1) on distraction osteogenesis in rabbits. A total of 61 rabbits were studied. Seven days after tibial osteotomy, distraction was started at a rate of 0.25 mm/12 h for 3 weeks. Starting with distraction, TGF-beta1 was applied in four different dosages (0, 10, 20, and 40 ng/day) at the site of osteotomy through a catheter connected to a subcutaneously implanted miniosmotic pump. Rabbits were killed at the end of the distraction period or 3 weeks later, and histological, densitometric, and biomechanical parameters were assessed. TGF beta1 treatment had no detectable effect on bone mineral density or histologically determined bone volume in the distraction gap but it increased the amount of fibrous tissue in the callus region. Load to failure in uniaxial tension tended to be lower in TGF-beta1-treated animals. In conclusion, TGF-beta1 treatment during distraction osteogenesis did not have a beneficial effect in this model. PMID- 10831935 TI - Expression of neurotrophins and their receptors (TRK) during fracture healing. AB - To clarify the roles of neurotrophins and their receptors in bone formation, expression of neurotrophins and their receptors (TRK) in a model of mouse fracture healing was investigated. A total of 120 male ICR mice were studied. The right eighth rib of 70 mice was fractured. For sham operation as a control, the right eighth rib of 50 mice was similarly exposed but not fractured. Localization of TRKA, TRKB, and TRKC in a rectangular region of the rib together with surrounding soft tissues was investigated by immunostaining. Localizations of nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and neurotrophin-3 (NT-3) at the fracture callus were also investigated by immunostaining, and their mitochondrial RNA (mRNA) expressions were investigated by reverse transcriptase-polymerase chain reaction (RT-PCR). As a result, we observed two types of neurotrophin receptors in the bone forming area: immunostaining by anti-TRKA was observed in almost all bone forming cells, and staining with anti-TRKC was observed in osteoblast-like cells and hypertrophic chondrocytes, but no staining was observed with anti-TRKB. On the other hand, localization of NGF was observed in almost all bone forming cells, localization of BDNF was observed in osteoblast-like cells, and localization of NT-3 was observed in osteoblast-like cells and hypertrophic chondrocytes at the fracture callus. Expression levels of the mRNA of three neurotrophins in the fractured rib were increased during the process of healing, especially those of NGF and NT-3, which peaked at 2 days after the fracture. The level of BDNF mRNA increased gradually over 8 days. These findings show that neurotrophins and their receptors were expressed in bone forming cells, and suggest that they are involved in the regulation of bone formation as an autocrine and paracrine factor in vivo. PMID- 10831936 TI - Low-megahertz ultrasonic properties of bovine cancellous bone. AB - Ultrasound offers a noninvasive means to detect changes that occur to the density of cancellous bone as a result of degenerative diseases such as osteoporosis. Techniques based on the velocity and frequency dependence of attenuation of ultrasonic pulses propagated through cancellous bone have proven sensitive to bone density. Most previous studies have investigated these two parameters in the frequency range of 0.1-1.0 MHz. The present study had two goals. The first was to measure three ultrasonic parameters: longitudinal mode velocity; broadband ultrasonic attenuation (BUA); and apparent integrated backscatter (AIB), at higher frequencies using a broadband 2.25 MHz measurement system. The second goal was to assess the dependence of these parameters on bone density. Twenty-one specimens of cancellous bone acquired from the proximal end of four bovine tibiae were investigated in this study. The apparent density of the specimens (determined with the bone marrow removed and the specimens thoroughly dry) ranged between 0.3 and 0.9 g/cm(3). Ultrasonic measurements were performed along three mutually perpendicular directions corresponding to the anteroposterior (AP), mediolateral (ML), and superoinferior (SI) axes of the tibia. A linear regression was used to analyze the results of these measurements as a function of apparent density. Velocity demonstrated a highly significant linear increase with density for all three directions (AP: p < 0.001; ML: p < 0.001; SI: p < 0.01). AIB decreased with density in all three directions; however, only the ML and SI directions demonstrated a significant linear correlation (AP: p = n.s.; ML: p < 0.05; SI: p < 0.05). In the frequency range 0.5-1.0 MHz, BUA exhibited a significant linear increase in the AP and ML directions, but not the SI direction (AP: p < 0.05; ML: p < 0.01; SI: p = n.s.). In contrast, in the frequency range 1.0-2.0 MHz, BUA exhibited a highly significant increase with density in the SI direction, but no significant change in the AP and ML directions (AP: p = n.s., ML: p = n.s., SI: p < 0.001). PMID- 10831937 TI - Parathyroid hormone and growth hormone have additive or synergetic effect when used as intervention treatment in ovariectomized rats with established osteopenia. AB - The severely osteoporotic human skeleton is characterized by thin cortices and a very fragile cancellous framework. To increase the biomechanical competence of such a skeleton, powerful anabolic agents are needed. The aim of the present study was to compare the effect of parathyroid hormone (PTH), growth hormone (GH) and combination treatment with PTH and GH in an aged, rat model with established osteopenia. Furthermore, envelope- and site-specific effects of the two agents are described. Twelve-month-old virgin F344 rats were divided into six groups with 11 animals per group: (1) baseline; (2) sham-operated + solvent vehicle (s.v.) (sham); (3) ovariectomized + s.v. (ovx); (4) ovx + GH 2.5 mg/kg body weight per day; (5) ovx + PTH 80 microg/kg body weight per day; and (6) ovx + GH and PTH treatment. Group 1 were killed to establish baseline values. Groups 2 (sham) and 3 (ovx) were killed after 24 weeks. Groups 4, 5, and 6 were allowed to develop osteopenia for 16 weeks before treatment was initiated. Treatment was given for a period of 8 weeks. The effects of GH, PTH, and GH + PTH cotherapy were measured by biomechanical testing at four different skeletal sites: lumbar vertebra; femoral diaphysis; femoral neck; and distal femoral metaphysis. In addition, static histomorphometry was performed at the middiaphyseal region. Ovx induced a loss of bone strength at all sites, but this was significant only at the femoral diaphysis and distal metaphysis. GH could reverse the loss of strength at the diaphysis, but not at the metaphysis. PTH, on the other hand, reversed the loss of strength to values significantly over ovx at all four sites. At the metaphysis, PTH monotherapy increased strength to above sham levels. However, GH + PTH cotherapy showed additive or synergistic effects at the four tested sites, leading to strength values significantly over sham at all these sites. Static histomorphometry showed that GH exerted its main effect on the periosteal envelope and PTH on the endocortical envelope; for this reason, the GH + PTH combination treatment had an additive or synergistic effect. We conclude that GH and PTH have a very pronounced anabolic effect when given in cotherapy. Therefore, this treatment regime seems promising in the clinical situation for management of patients with severe, established osteoporosis. PMID- 10831938 TI - Coordinated cytokine expression by stromal and hematopoietic cells during human osteoclast formation. AB - An in vitro culture system to generate human osteoclasts (OC) was recently described in which OC precursors in the human peripheral blood mononuclear cell (PBMC) population differentiate in the presence of murine ST-2 stromal cells. We used this culture system to define the cytokine environment in which human OC form and to determine the separate contributions of the stromal and hematopoietic elements. We designed a panel of reverse transcriptase-polymerase chain reaction (RT-PCR) primers that specifically amplify the respective murine or human mRNA species that correspond to cytokines and their cognate receptors previously shown to promote or inhibit OC differentiation. ST-2 cells were cocultured with human PBMC for up to 21 days in the presence of 1alpha,25(OH)(2) vitamin D(3), dexamethasone, and recombinant human macrophage-colony stimulating factor (M CSF). OC formation was monitored by the appearance of cells that were positive for tartrate-resistant acid phosphatase (TRAP) and able to form resorption lacunae on slices of dentine. We found that the ST-2 cells in these cultures express messenger RNA (mRNA) encoding a repertoire of many of the reported osteoclastogenic factors (interleukins [IL]-1/IL-1R1, IL-11, IL-6/IL-6R, and IL 17 transforming growth factor [TGF]-beta), as well as the recently described OC differentiation factor (ODF/TRANCE/RANKL). The stromal cells also expressed mRNA encoding two molecules shown to be inhibitory to osteoclastogenesis, osteoprotegerin (OPG) and IL-18. OPG, IL-1, IL-1R1, IL-6, IL-6R, IL-11R, IL-17, IL-18, IL-18R, TGF-beta, and M-CSF were expressed by both the stromal cells and the PBMC. Expression of mRNA encoding RANK, IL-1R2, and c-fms, was specific for the PBMC. In addition, PBMC were found to express sIL-6R, granulocyte macrophage (GM)-CSF, GM-CSFRalpha, and tumor necrosis factor (TNF)-alpha. Whereas this indicated that human OC formation occurs in a complex environment of many positive and negative influences, we identified three apparent features of the cytokine environment that may be a characteristic of normal osteoclast formation. First, the ratio of mouse ODF:OPG mRNA was found to increase during the cocultures, consistent with a key role for ODF in the promotion by stromal cells of OC formation. Second, we found that mRNA encoding IL-1 and IL-17, as well as IL-6 and sIL-6R, were coordinately expressed by the PBMC. Third, analysis of the culture medium showed that the PBMC secreted IL-1, IL-6, and TNF-alpha protein only in coculture with ST-2 cells during the first few days of osteoclast development. We conclude that human OC formation occurs in a complex environment of many positive and negative influences; however, these are likely to be strictly regulated by a coordinated cytokine response of both stromal and hematopoietic cells. PMID- 10831939 TI - Inferior alveolar nerve transection inhibits increase in osteoclast appearance during experimental tooth movement. AB - To evaluate the role of sensory nerve innervation in alveolar bone remodeling during experimental tooth movement, we investigated histomorphometrically the influence of sensory nerve denervation on bone metabolism. Seven days after inferior alveolar nerve (IAN) transection or a sham operation in rats, orthodontic force was applied to the animals by inserting an elastic module interproximally between the lower first molar and second molar. Twenty-four hours after the application of the orthodontic force, osteoclast number, osteoclast surface, and osteoblast surface were measured on the trabecular bone surface in the interradicular septum of the lower second molar. The distribution of sensory nerve fibers immunoreactive to antibody against calcitonin gene-related peptide (CGRP) was also evaluated. In the sham-operated rats, CGRP-immunoreactive nerves were observed to be distributed along the blood vessels in the trabecular alveolar bone. Experimental tooth movement resulted in a fivefold increase in the number of osteoclasts and in increased immunoreactivity of nerves to anti-CGRP in the trabecular bone. However, IAN transection depleted the immunoreactivity to anti-CGRP and reduced the osteoclast number and osteoclast surface significantly. On the other hand, in the rats that were not subjected to experimental tooth movement, there was no significant difference in osteoclast number between sham operated and IAN-transected rats. Significant changes were not observed in osteoblast surfaces associated with experimental tooth movement or nerve transection. These findings suggest that sensory nerves play an important role in regulating bone resorptive activity during experimental tooth movement. PMID- 10831940 TI - Comparison of sexual activity of women and men after a first acute myocardial infarction. AB - Studies of gender differences in the sexual activity of men and women after a first acute myocardial infarction (AMI) have produced conflicting results. The present study was performed to determine whether there are gender differences (1) in the quantity and quality of sexual activity after a first AMI, and (2) in the relations between selected demographic and medical variables and sexual activity after AMI. Four hundred sixty-two men and 51 women with a first AMI were interviewed once before discharge and again 3 to 6 months after AMI. Patients' demographic and medical background and their frequency of and satisfaction with sexual behavior were obtained from the interviews and from medical charts. Analyses of variance showed that women reported significantly less frequency of and satisfaction with sexual activity than men before and after AMI. Both women and men reported significantly less sexual activity and less satisfaction with sexual activity after AMI than before AMI. The decrease in frequency of and satisfaction with sexual activity after AMI was similar for women and men. The relations between selected demographic and medical variables such as age, education, and perceived health before the first AMI and the frequency of and satisfaction with sexual activity of the women and men did not appear to be affected differently by the AMI. A first AMI appears to reduce the frequency of and satisfaction with sexual activity of women and men similarly 3 to 6 months after AMI. PMID- 10831941 TI - Circulating monocytes in patients with diabetes mellitus, arterial disease, and increased CD14 expression. AB - Low serum concentrations of high-density lipoprotein (HDL) cholesterol and elevated levels of acute-phase reactans are frequently found in patients with non insulin-dependent diabetes mellitus (NIDDM) and cardiovascular disease. Changes in the phenotype of circulating monocytes have been reported with both of these circumstances in nondiabetic subjects. In the present study, we explored the possibility that similar changes may occur in circulating monocytes of patients with NIDDM and arterial disease. Two groups of subjects with NIDDM were studied: patients with cardiovascular disease (n = 25) were compared with a group without cardiovascular disease (n = 26); both groups were age- and sex-matched, had the same length of diabetes duration, and degree of glycemic control. Healthy nondiabetic volunteers of comparable age and sex (n = 35) formed the control group. There was no significant difference in the numbers of the CD14+/CD16+ monocyte subpopulations between the 3 groups. However, a significant graded increase of the mCD14 intensity expression values was observed among the groups, with the highest levels in patients with NIDDM patients and the lowest in nondiabetic subjects. The serum C-reactive protein concentrations were significantly higher in the group with arterial disease compared with those without arterial disease or healthy controls. In the group of patients as a whole, relative mCD14 intensity expression was significantly correlated with HDL cholesterol levels (inversely) and with serum concentrations of C-reactive protein. Serum HDL cholesterol levels and the C-reactive protein concentrations were also significantly correlated. We concluded that the increased mCD14 intensity expression on circulating monocytes may be an important contributor to the increased inflammatory response observed in patients with NIDDM and arterial disease, and eventually, to atherogenesis. PMID- 10831942 TI - Role of cardiac surgery in the hospital phase management of patients treated with primary angioplasty for acute myocardial infarction. AB - Although cardiac surgery is performed in approximately 10% of acute myocardial infarction (AMI) patients undergoing a primary percutaneous transluminal coronary angioplasty (PTCA) reperfusion strategy before discharge, the indications for and timing of operative revascularization, and the short- and long-term outcomes after surgery have not been characterized. In the prospective, controlled Primary Angioplasty in Myocardial Infarction-2 trial, cardiac catheterization was performed in 1,100 patients within 12 hours of onset of AMI at 34 centers, followed by primary PTCA when appropriate. Cardiac surgery was performed before hospital discharge in 120 patients (10.9%), electively in 42.6%, and on an urgent or emergent basis in 57.4%. Surgery was performed in 6.1% of 982 patients after primary PTCA (although emergently for failed PTCA in only 4 cases [0.4%]), and in 53 of 118 patients (44.9%) not undergoing primary PTCA. Patients requiring surgery were older, and more frequently had diabetes and 3-vessel disease than those managed nonoperatively. Internal mammary artery grafts were placed in only 31% of patients. In-hospital mortality was 6.4% in patients undergoing urgent/emergent surgery, 2.0% after elective surgery, and 2.6% in patients not undergoing surgery (p = NS). After multivariate correction for baseline risk factors, early and late survival free of reinfarction were similar in patients undergoing versus not undergoing in-hospital cardiac surgery. Thus, the appropriate use of coronary artery bypass graft surgery in the peri-infarction period is an integral component of the primary PTCA approach, and is frequently used to optimize the prognosis of a high-risk AMI cohort with unfavorable baseline features. The implications for the performance of primary PTCA in AMI at centers without on-site surgical facilities are discussed. PMID- 10831943 TI - Phytoestrogens do not influence lipoprotein levels or endothelial function in healthy, postmenopausal women. AB - Plant estrogen or phytoestrogens (PE) are increasingly consumed for the purposes of menopause symptom relief and prevention of cardiovascular and other diseases. The objective of this study was to evaluate the effects of PE on plasma lipids and lipoproteins and on endothelial function. Twenty healthy, postmenopausal women, 50 to 70 years old, and with evidence of endothelial dysfunction, were treated with a soybean PE tablet of 80 mg/day of isoflavones. Endothelial function was assessed noninvasively using brachial ultrasound. A double-blind, placebo-controlled, randomized crossover design was employed. After 3 weeks stabilization on a standard fat-reduced diet, subjects received PE or placebo for 8 weeks in random order, separated by a washout period of 8 weeks. Compared with placebo, there were no significant effects of PE on blood pressure and plasma lipid or lipoprotein concentrations. Flow-mediated endothelium-dependent dilation (FMD) in response to reactive hyperemia was not significantly changed by PE ingestion (3. 3 +/- 0.7% on placebo vs 4.1 +/- 0.7% on PE, p >0.4). Variation in FMD was not correlated with change in plasma isoflavone concentration (r = -0.09, p >0.7). Glyceryl trinitrate endothelium-independent dilation was not significantly changed with PE (15.9 +/- 1.3% vs 13.7 +/- 1.2%, p >0.1). These results fail to show a significant impact of medium-term supplementation with 80 mg/day of isoflavones on lipid and lipoprotein levels or on endothelial function in healthy, postmenopausal women. PMID- 10831944 TI - Randomized comparison of two targets in typical atrial flutter ablation. AB - Typical atrial flutter ablation has become anatomically guided to 2 separate sites within the isthmus at the inferior right atrium: (1) between the inferior vena cava and the tricuspid annulus (anterior side of the isthmus [A]), (2) between the eustachian crest, the coronary sinus ostium and tricuspid annulus (posterior side of the isthmus [P]). We prospectively compared ablation results at these sites in 72 consecutive patients. Patients were randomized in group P or A according to the initial target site. If ablation failed at 1 site after 15 radiofrequency (RF) pulses, the other side of the isthmus was targeted. Before 15 RF pulses, complete bidirectional isthmus block was achieved in 30 of 36 group A patients and in 25 of 36 group P patients, with similar mean RF pulses number, procedure time, and fluoroscopy time. After shifting to the other target, success was finally obtained at P in 2 of 6 group A patients, and at A in 8 of 11 group P patients before a maximum of 30 RF pulses. Among successful patients, number of RF pulses, procedure time, and fluoroscopy time were significantly lower in group A (7.2 +/- 5.4 vs 11.0 +/- 8.1 pulses, p = 0.03; 131 +/- 44 vs 163 +/- 66 minutes, p = 0.03; 31 +/- 19 vs 46 +/- 24 minutes, p = 0.01, respectively). Impairment of atrioventricular (AV) nodal conduction occurred in 5 patients only during ablation at P. AV block was transient in 4 patients and permanent in 1. Although atrial flutter ablation is equally effective at P and A, success seems easier to obtain when A is first targeted. Ablation at P is associated with a significant risk of AV block. PMID- 10831945 TI - Late results of percutaneous mitral commissurotomy for calcific mitral stenosis. AB - The aim of this study was to assess late results of percutaneous mitral commissurotomy (PMC) in calcific mitral stenosis and to identify predictors to improve patient selection. We analyzed 422 patients who underwent PMC for calcific mitral stenosis. The extent of calcium was graded from 1 to 4 by fluoroscopy: 227 patients (53%) were graded 1, 125 (30%) graded 2, 55 graded 3 (13%), and 15 graded 4 (4%). The procedure failed in 15 patients, used a single balloon in 11, a double balloon in 126, and the Inoue balloon in 270. In-hospital mortality was 1.2%. Good immediate results (valve area >/=1.5 cm(2) without mitral regurgitation >2/4), were obtained in 321 patients (76%). Multivariate analysis identified 5 predictors of good immediate results: a younger age (p = 0.0004), a lesser degree of stenosis (p = 0.0005), a smaller extent of calcium (p = 0.04), the use of the Inoue balloon (p = 0.015), and a larger effective balloon dilating area (p = 0.006). Good functional results, defined as survival with no further intervention and in New York Heart Association class I or II, were 36 +/- 4% at 8 years. The predictors of good functional results after good immediate results were a younger age (p = 0.04), a lower pre-PMC New York Heart Association class (p <0.0001), sinus rhythm (p = 0.0006), a smaller extent of calcium (p = 0.02), and a lower gradient after PMC (p <0.0001). Despite a frequent deterioration on follow-up after PMC for calcific mitral stenosis, the predictive analysis suggests that PMC may be useful in deferring surgery in selected patients with mild to moderate calcific deposits, who have otherwise favorable characteristics. PMID- 10831946 TI - Single-strand conformation polymorphism analysis on the delta-sarcoglycan gene in Japanese patients with hypertrophic cardiomyopathy. AB - To elucidate the etiology of hypertrophic cardiomyopathy (HC) in humans, we analyzed the delta-sarcoglycan gene (SG), which is reported to be the causal gene for HC in the Syrian hamster BIO14.6. We performed polymerase chain reaction (PCR) single-strand conformation polymorphism (SSCP) and nucleotide sequence analyses on the delta-SG in 102 patients with HC. SSCP was detected in exon 2 of the gene, but not in the other exons. The direct sequencing analysis of exon 2 revealed a C-->T substitution at nucleotide residue 84 (TAC-->TAT) with no amino acid alteration (Tyr-->Tyr). There were no significant differences in allele frequencies of C/T between the patients with HC and the control group. Patients with HC were classified into 4 subgroups: obstructive HC, nonobstructive HC, apical HC, and familial HC. The allele frequency of C/T polymorphism in each of these groups was compared with that of the control group. The obstructive HC group showed a significantly greater frequency of the allele T than in the control group (31.6% vs 15.1%, RR = 2.6, p = 0.023). No other significant differences were observed. Thus, amino acid alteration in delta-SG may not be a common cause of HC in Japanese patients. PMID- 10831947 TI - Echocardiographic predictors of coronary artery pathology in pulmonary atresia with intact ventricular septum. AB - Coronary artery pathology is a major determinant of treatment strategy and outcome in patients with pulmonary atresia and intact ventricular septum (PA/IVS). For this reason, infants with PA/IVS routinely undergo preoperative cardiac catheterization. The goal of this study was to identify echocardiographic predictors of coronary artery pathology in infants with PA/IVS. The initial preoperative echocardiograms of 30 consecutive infants with PA/IVS (median age at diagnosis 1 day) were reviewed for indexes predicting the degree of coronary pathology. The tricuspid valve (TV) annulus diameter Z- score was determined and evidence of abnormal flow in the coronary arteries by Doppler was evaluated. Coronary pathology was defined by angiography and graded as: 0 = no fistulae; 1 = fistulae/no right ventricular (RV)-dependent coronary arteries; 2 = fistulae with 1 RV-dependent coronary; 3 = fistulae with >/=2- vessel RV-dependent coronary arteries. Outcome was classified as: 2 ventricles, "1.5" ventricles, and 1 ventricle. By angiography, 30% of the patients had grade 0 coronary pathology, 30% had grade 1, 20% had grade 2, and 20% had grade 3. There was 1 death in a patient with grade 3 coronary pathology. Among the survivors (median age at follow-up 28. 6 months), biventricular circulation existed in 12 patients (41%), 7 patients (24%) were 1.5, and 10 (34%) were 1 ventricle. All patients with TV Z score -2.5. The sensitivity, specificity, positive, and negative predictive values of TV Z-score 0.5 in all group 1 infants, and in 6 of 13 group 2 infants (2 sepsis deaths, 4 palliations). Compared with a tricuspid valve z-score >-3, a tricuspid/mitral ratio >0.5 was a better predictor of biventricular repair. Thus, infants who have a successful biventricular repair have significantly greater preoperative weight, tricuspid valve z-scores, and tricuspid/mitral valve ratios. A tricuspid/mitral ratio >0.5 was the best predictor of a biventricular repair. PMID- 10831949 TI - Orthotopic heart transplantation: standard versus bicaval technique. AB - We compared orthotopic heart transplantation (HT) by bicaval technique with the standard technique. Between January 1995 and December 1997, 117 patients underwent 118 HTs; 71 patients (15 women and 56 men) had 72 HTs by standard technique and 46 patients (9 women, 37 men) underwent HT using bicaval procedures. Preoperative parameters were similar in both groups; 5 patients who underwent the standard technique and no patients who underwent bicaval procedures required permanent pacemakers (p = NS). Isoproterenol infusion was significantly longer in the standard technique. Major perioperative arrhythmias (ventricular tachycardia and fibrillation, asystole) appeared in 8.2% and 7.0% of standard and bicaval HTs, respectively; atrial fibrillation appeared in 13.1% and 4.6%, respectively (p = NS). At 1 month, mitral and tricuspid regurgitation rates were higher in the standard group (p = NS); at 1 year only tricuspid regurgitation was still higher (p = NS). Right atrial pressure, Wood units, cardiac output, and cardiac index were examined (p = NS). At multivariate analysis, interaction between preoperative Wood units and transplant type was elicited for Wood units at 1 month and for right atrial pressure at 1, 3, and 6 months. In the high resistance subgroup, the patients who underwent bicaval procedures had higher resistances at 1 month. In the low resistance subgroup, right atrial pressure was higher in patients who underwent standard techniques at 1, 3, and 6 months follow up. Thus, bicaval HT was found to be safe, without surgically related complications, it provoked significantly less blood loss, and required less isoproterenol use. No significant advantages were observed in conduction disturbances and major arrhythmias or regarding the need for temporary or permanent pacemakers. PMID- 10831950 TI - Risk of pulmonary embolism and/or deep venous thrombosis in Asian-Americans. AB - Several reports from Asian countries suggest a low prevalence of pulmonary embolism (PE) and deep venous thrombosis (DVT) in Asians, and sparse US data show that a slightly higher prevalence of PE/DVT in "nonwhites" than in whites is evident in all geographic regions except the Pacific region (California, Oregon, and Washington) where "nonwhites" include a larger proportion of Asians and Hispanics than in other US locations. We prospectively studied PE/DVT hospitalizations in 128,934 persons in relation to traits determined at health examinations in 1978 to 1985. Through 1994, 337 persons were subsequently hospitalized for PE and/or DVT (for PE first, n = 206). Cox proportional-hazards models with 9 covariates were used. In multivariate models, the following RRs (95% confidence intervals) were found for PE/DVT combined: black/white = 1.1 (0.4 to 1.4); Hispanic/white = 0.7 (0.3 to 1.5); and Asian/white = 0.2 (0.1 to 0. 5; p = 0.002). The lower risk of Asians was present in each sex and for persons first hospitalized for either PE or DVT. Covariates with significant positive relations to risk were age, male sex, body mass index, and a composite coronary disease risk/symptom variable; covariates not significantly related were education, marital status, smoking, and alcohol. These data suggest that Asians have very low risk of PE/DVT, which may account for US geographic variations in white/non white risk differences. Possible explanations include the absence of hazardous mutations or unspecified PE/DVT protective traits in Asians. PMID- 10831951 TI - Effect of risk factors on the costs of coronary surgery. PMID- 10831952 TI - Francis Robicsek, MD, PhD: a conversation with the editor. Interview by William Clifford Roberts, MD. PMID- 10831953 TI - Electron beam tomography comparison of culprit and non-culprit coronary arteries in patients with acute myocardial infarction. PMID- 10831954 TI - Left ventricular function during combined isometric and dynamic exercise-induced ST depression in coronary artery disease. PMID- 10831955 TI - Usefulness of exercise tomographic myocardial perfusion imaging for detection of restenosis after coronary stent implantation. PMID- 10831956 TI - Comparison of clinical outcome following coronary stenting or balloon angioplasty in dialysis versus non-dialysis patients. PMID- 10831957 TI - Prevalence of patent foramen ovale in patients with acute myocardial infarction and angiographically normal coronary arteries. PMID- 10831958 TI - Coronary interventional procedures in pediatric heart transplant recipients with cardiac allograft vasculopathy. PMID- 10831959 TI - Ablation of right and left atrial premature beats following cardioversion in patients with chronic atrial fibrillation refractory to antiarrhythmic drugs. PMID- 10831960 TI - Hypersensitivity of cerebral artery response to catecholamine in patients with neurally mediated syncope induced by isoproterenol. PMID- 10831961 TI - Effect of sensing system on the incidence of myopotential oversensing during bradycardia pacing in implantable cardioverter-defibrillators. PMID- 10831962 TI - Prognosis of congestive heart failure after prior myocardial infarction in older men and women with abnormal versus normal left ventricular ejection fraction. PMID- 10831963 TI - Prediction of peak oxygen uptake from cycle exercise test work level in heart failure patients >/=65 years of age. PMID- 10831964 TI - Transesophageal echocardiographic results of catheter closure of atrial septal defect in children and adults using the Amplatzer device. PMID- 10831965 TI - Relation between left ventricular and/or left atrial thrombus and anticardiolipin antibodies in patients with acute myocardial infarction. PMID- 10831966 TI - Plasma brain natriuretic peptide in athletes. PMID- 10831967 TI - Hysterectomy outcomes in patients with similar indications. AB - OBJECTIVE: To investigate the cost advantages and complication rates associated with surgical routes of uncomplicated hysterectomies in which uteri weigh less than 280 g and benign diseases are confined to the uterus. METHODS: Data were collected prospectively from 1988 to 1993 from 4609 consecutive women who had hysterectomies at a single institution. Women who had abdominal hysterectomies, laparoscopically assisted vaginal hysterectomies, or vaginal hysterectomies were selected if they had benign diseases confined to the uterus (adenomyosis, leiomyomas, abnormal uterine bleeding, cervical carcinoma in situ, and prolapse) and uterine weights less than 280 g. We compared length of stay, hospital charges, and associated complications between groups. RESULTS: A total of 1427 women met the study criteria. Length of stay was longer after abdominal hysterectomies than laparoscopically assisted vaginal hysterectomies or vaginal hysterectomies (3.99 +/- 1.16 days, 2.45 +/- 1.58 days, and 2.76 +/- 0.94 days, respectively; P <.001). Hospital charges for vaginal hysterectomies were significantly lower than for either abdominal or laparoscopically assisted vaginal hysterectomies (P <.001). The median charge for vaginal hysterectomies was $4166; the median charges for laparoscopically assisted vaginal hysterectomies and abdominal hysterectomies were 71% and 35% higher than this, respectively. There was a higher risk of one or more complications after abdominal hysterectomies (9.3%) than after laparoscopically assisted vaginal hysterectomies (3.6%; P <.001) or vaginal hysterectomies (5.3%; P <.001). The incidence of postoperative infection or fever was higher after abdominal than after vaginal hysterectomies (4.0% versus 0.8%; P =.029). CONCLUSION: This study supports the vaginal route of hysterectomy when disease is confined to the uterus and uterine weight is less than 280 g. PMID- 10831968 TI - Intraoperative cystoscopy in conjunction with anti-incontinence surgery. AB - OBJECTIVE: To determine the frequency of lower urinary tract injury detected by routine intraoperative cystoscopy after anti-incontinence surgery. METHODS: We reviewed charts from women who had anti-incontinence surgery and routine intraoperative cystoscopy done by a single surgeon from June 1, 1995, to June 1, 1998, and assessed preoperative and intraoperative variables. RESULTS: We reviewed 351 patient records. Four records were incomplete and there were nine injuries in the other 347 cases (2.6%, 95% confidence interval [CI] 1.2, 4.9). Four cystotomies occurred during laparoscopic Burch procedures and were detected before cystoscopy. Five injuries were detected at cystoscopy, a rate of 1.5% (95% CI 0. 5, 3.4). Four injuries occurred during 161 pubovaginal sling procedures (2.5%, 95% CI 0.7, 6.2). One woman had sutures in her bladder from a prior procedure detected at cystoscopy. In 186 Burch procedures (48 laparoscopic, 138 open), there were no previously unrecognized injuries detected by cystoscopy. All injuries were repaired during original surgery. It was not possible to assess preoperative and intraoperative risk factors because of the low rate of injury. CONCLUSION: The rate of injury to the lower urinary tract during anti incontinence surgery in this series was 2.6% (95% CI 1.2, 4.9). Injuries during Burch procedures were all detected before cystoscopy. PMID- 10831969 TI - Quantification of intramuscular nerves within the female striated urogenital sphincter muscle. AB - OBJECTIVE: To analyze the quantity and distribution of intramuscular nerves within the striated urogenital sphincter and test the hypothesis that decreased nerve density is associated with decreased striated sphincter muscle and cadaver age. METHODS: Thirteen cadaveric urethras (mean age 47 years, range 15-78 years) were selected for study. A sagittal histologic section was stained with S100 stain to identify intramuscular nerves. The number of times that a nerve was seen within the striated urogenital sphincter (nerve number) was counted. The number of axons within each nerve fascicle was also counted. Regression analysis of nerve density against muscle cell number and age was performed. RESULTS: Remarkable variation was found in the quantity of intramuscular nerves in the striated urogenital sphincter of the 13 urethras studied. The number of nerves ranged from 72 to 543, a sevenfold variation (mean 247.1 +/- standard deviation 123.2), and the range of number of axons was 431 to 3523 (2201 +/- 1152.6). The larger nerve fascicles were seen predominantly in the distal (13.1 +/- 5.7 axons per nerve) compared with the proximal part of the striated urogenital sphincter (1.2 +/- 2). Reduced nerve density throughout the striated urogenital sphincter correlated with fewer muscle cells (P =.02). Nerve density also decreased with advancing age (P =.004). CONCLUSION: Remarkable variation in the quantity of intramuscular nerves was found. Women with sparse intramuscular nerves had fewer striated muscle cells. Intramuscular nerve density declined with age. PMID- 10831970 TI - Endocervical curettage when colposcopic examination is satisfactory and normal. AB - OBJECTIVE: To estimate the incidence of endocervical dysplasia in women with cervical cytology of atypical squamous cells of undetermined significance (ASCUS) or low-grade squamous intraepithelial lesion (SIL) who have a satisfactory and normal colposcopic examination. METHODS: An electronic colposcopy database was reviewed and women with satisfactory colposcopic examinations and original cervical cytology of ASCUS on two consecutive Papanicolaou smears, ASCUS favor SIL, or low-grade SIL were selected. Exclusion criteria were pregnancy, insufficient endocervical curettage (ECC), or colposcopic examination that showed an abnormality that required cervical biopsy. Subjects also were excluded if they were postmenopausal or had surgical or ablative therapy for cervical dysplasia within the past year. A computerized review of 2517 patient records found 860 that met the search criteria. A manual review of those records using the exclusion criteria isolated a study group of 159 women. RESULTS: Four of 159 subjects (2.5%, 95% confidence interval [CI] 0.69, 6.3) had dysplastic cells in endocervical curettings. In these four, the ECC specimens had benign endocervical cells and separate fragments of squamous cells with mild dysplasia. In three women, loop electrosurgical excision procedures showed mild dysplasia limited to the transformation zone. The fourth subject was believed to have contamination from an unrecognized ectocervical lesion and was treated conservatively. A repeat ECC found benign endocervical cells. Involvement of the endocervix by dyplasia was excluded in all but one of 159 patients (0.63%, 95% CI 0.02, 3.5). CONCLUSION: Incidence of endocervical dysplasia was extremely low in women with cervical cytology of consecutive ASCUS, ASCUS favor SIL, or low-grade SIL who have a satisfactory and normal colposcopic examination. Our findings suggest that endocervical curettage might be safely avoided in those women. PMID- 10831971 TI - Hormone replacement therapy for oxidative stress in postmenopausal women with hot flushes. AB - OBJECTIVE: To assess the association of hot flushes during postmenopause with oxidative stress and to determine whether hormone replacement therapy (HRT) affects the plasma redox status of postmenopausal women. METHODS: We conducted a prospective clinical study of 49 postmenopausal women who have (n = 29) or do not have (n = 20) hot flushes. Twelve of the postmenopausal women with hot flushes and six without were treated with HRT (estradiol patches and medroxyprogesterone acetate) for 4 months. Plasma level of estradiol, total antioxidant status, reduced sulfhydryl groups, lipoperoxides, total cholesterol, and triglycerides were measured at 4-month intervals in both groups, before and after treatment. RESULTS: Postmenopausal women who have hot flushes, had lower total basal antioxidant status in plasma (.9 +/-.01 compared with 1.14 +/-.01 mmol/L), lower concentration of reduced sulfhydryl groups (145 +/- 4 compared with 200 +/- 3 micromol/L), and higher concentration of lipoperoxides (2.88 +/-.04 compared with 2.61 +/-.04 micromol/L) than women without hot flushes. After HRT, total antioxidant status and reduced sulfhydryl groups increased, and lipoperoxides decreased similarly in both groups. Hormone replacement therapy decreased the frequency of hot flushes per day from 11.2 +/- 0.8 to 1.4 +/- 0.3. CONCLUSION: Hot flushes in postmenopausal women were associated with the oxidative process. Hormone replacement therapy decreases oxidative stress and the number of episodes of hot flushes. Because oxidative stress is associated with a high risk for cardiovascular diseases, HRT might protect women with hot flushes. PMID- 10831972 TI - A randomized comparison of danazol and leuprolide acetate suppression of serum soluble CD23 levels in endometriosis. AB - OBJECTIVE: To determine the effects of treatment with danazol and leuprolide acetate depot on serum-soluble CD23 concentrations in women with endometriosis. METHODS: This randomized trial involved 20 women 18-42 years old with regular menses and known pelvic endometriosis who were recruited from a university hospital between 1993 and 1998. Ten women took 200 mg of danazol three times daily for 6 months, and the remaining ten were given 3.75 mg of leuprolide acetate depot every 28 days for 6 months. Blood-soluble CD23 levels were measured before treatment, during the last 15 days of the 6-month treatment course, and 3 months after treatment. Only one blood sample was taken from ten women without endometriosis, between the 5th and 7th days of their menstrual cycles. For statistical analysis, we used independent and paired t tests with the Pearson correlation coefficient. RESULTS: Soluble CD23 levels were significantly higher in women with endometriosis before treatment than in ten normal controls. Levels decreased significantly during treatment with either danazol or leuprolide acetate. Three months after treatment, soluble CD23 values remained lower than before treatment. There was no correlation between soluble CD23 concentrations and severity of endometriosis. CONCLUSION: Our findings suggest that endometriosis increases soluble CD23 levels, which can be suppressed with either danazol or leuprolide acetate injection. PMID- 10831973 TI - Risk factors for diethylstilbestrol-associated clear cell adenocarcinoma. AB - OBJECTIVE: To assess the influence of postnatal factors on the development of clear cell adenocarcinoma in women exposed to diethylstilbesterol (DES), particularly factors related to exogenous or endogenous hormone exposures, and to reassess the relation of season of birth. METHODS: For the analysis of postnatal factors, 244 cases were compared with 244 age-matched DES-exposed women. Information was obtained from telephone interviews, with questions asked in reference to an index age, the age at which clear cell adenocarcinoma was diagnosed. For the analysis of season of birth, 604 cases, living or deceased, were compared with 1749 DES-exposed women. RESULTS: Neither oral contraceptive (OC) use nor pregnancy was associated with risk of clear cell adenocarcinoma: the odds ratios (OR) were 1.1 (95% confidence interval [CI] 0.7, 1.8) for OC use and 1.3 (95% CI 0.7, 2.3) for pregnancy. The OR for fall season of birth relative to all other seasons was 1.2 (95% CI 0.9, 1.4). CONCLUSION: Pregnancy and OCs do not increase risk of DES-associated clear cell adenocarcinoma. In addition, season of birth does not appear to be a material factor; the previous finding of an increased risk for fall season of birth is most likely not valid and probably a chance finding due to small sample size. PMID- 10831974 TI - Human papillomavirus testing in women with mild cytologic atypia. AB - OBJECTIVE: To evaluate the efficiency of human papillomavirus (HPV) testing by Hybrid Capture II (Digene Diagnostics Inc., Silver Spring, MD) with regard to detecting biopsy-confirmed cervical intraepithelial neoplasia (CIN) or high-grade CIN in women with mild atypia, compared with the efficiencies of polymerase chain reaction (PCR), Southern blot hybridization, and cytology. METHODS: We prospectively studied 378 women with atypical squamous cells of undetermined significance (ASCUS) (n = 111) or low-grade squamous intraepithelial lesions (SILs) (n = 267) demonstrated by referral cytology. We did repeat cytology, sampling for detection of HPV DNA by Hybrid Capture II, PCR, and Southern blot hybridization, and colposcopic evaluation with cervical biopsies. RESULTS: All participants underwent the Hybrid Capture II test and 320 underwent the three HPV tests. Sensitivities of Hybrid Capture II for detecting CIN and high-grade CIN (0.81 and 0.86, respectively) were similar to those of cytology (0.83 and 0.82, respectively) and PCR (0.77 and 0.95, respectively), and higher than those of Southern blot hybridization (0.48 and 0.45, respectively). Compared with cytology, combined triage with Hybrid Capture II improved sensitivities for detecting CIN (0.94 versus 0.83, P <.001) and high-grade CIN (0.96 versus 0.85), though the latter difference was not significant (P =.17). In women with ASCUS, sensitivities of combined triage and cytology for detecting CIN were 0.94 and 0.71, respectively (P =.01), and sensitivities of the two methods for detecting high-grade CIN were 0.92 and 0.66, respectively (P =.13). The increase in sensitivity was lower among women with low-grade SILs; for these women, cytology had high sensitivity (0.86 for CIN and 1.00 for high-grade CIN). The specificity of combined triage was significantly lower than that of cytology in both groups. CONCLUSION: Compared with repeat cytology, combined triage with HPV testing markedly improves sensitivity for detecting CIN in women with ASCUS, but at the expense of specificity. PMID- 10831975 TI - Cervical intraepithelial neoplasia outcomes after large loop excision with clear margins. AB - OBJECTIVE: To identify risk factors for residual or recurrent cervical intraepithelial neoplasia (CIN) after large loop excision of the transformation zone with clear margins. METHODS: We did a case-control study of women treated with loop excision for CIN who had adequate follow-up and in whom margins were believed to be clear. Women with clear margins in whom no subsequent lesions were found (controls) were compared with women who presented with subsequent CIN (cases). Epidemiologic and colposcopic risk factors for recurrence were analyzed. Multiple logistic regression analysis was done to identify independent risk factors. RESULTS: In 31 of 635 women studied (4.9%), subsequent lesions were diagnosed. Univariate analysis identified glandular involvement, satellite lesions, and age over 40 years in cases as significant. Multiple logistic regression analysis confirmed that these three characteristics were independent risk factors, with odds ratios of 4.9 (95% confidence intervals 1.9, 12.3), 19 (7.5, 48.2), and 6.7 (2.8, 15.8), respectively. Subsequent lesions were identified by colposcopy or cytologic testing during the first postoperative year in all but one case. CONCLUSION: Age over 40 years, glandular involvement, and satellite lesions were related to the reappearance of CIN after loop excision with clear margins. These findings could be used to define appropriate follow-up protocols. PMID- 10831976 TI - Cervical cancer diagnosed shortly after pregnancy: prognostic variables and delivery routes. AB - OBJECTIVE: To compare the prognoses of women diagnosed with cervical cancer during pregnancy with the prognoses of those diagnosed within 6 months after delivery and to assess the effect of vaginal delivery on recurrence risk and prognosis. METHODS: A matched case-control study of women with cervical cancer diagnosed during pregnancy or within 6 months of delivery was performed. Fifty six women had cervical cancer diagnosed during pregnancy and 27 within 6 months after delivery. Controls (cervical cancer diagnosed at least 5 years since last delivery) were matched one-to-one with cases based on age, histology, stage, treatment, and time of treatment. RESULTS: Among postpartum women, four had stage IA disease, 15 had stage IB1 or IB2, and eight had stage IIA or higher disease. Eleven had radical hysterectomies and 14 had radiation therapy. Two with stage IA1 disease were treated with vaginal hysterectomies. One of seven patients who had cesareans developed a local and distant recurrence. In contrast, ten of 17 (59%) who delivered vaginally developed recurrences (P =.04). In multivariate analysis, vaginal delivery was the most significant predictor of recurrence (odds ratio [OR] 6.91; 95% confidence interval [CI] 1.45, 32.8), followed by high stage (OR 4.66; 95% CI 1.05, 20.8). The survival for patients diagnosed in the postpartum period was significantly worse than for controls. CONCLUSION: Women diagnosed postpartum had worse survival than those diagnosed during pregnancy and were at significant risk of recurrent disease, particularly if they delivered vaginally. Therefore, pregnant women with cervical cancer should be delivered by cesarean. PMID- 10831977 TI - Borderline tumors of the ovary: correlation of frozen and permanent histopathologic diagnosis. AB - OBJECTIVE: To evaluate the correlation between the diagnosis of borderline tumor of the ovary by frozen and permanent pathology. METHODS: All pathology reports with diagnoses of borderline tumor of the ovary between 1980 and 1998 at Massachusetts General Hospital were reviewed. Univariate and multivariable logistic regression models were constructed for patient age, tumor size, histology, presence of bilateral or extraovarian disease, and concurrent diagnosis of endometriosis or endosalpingiosis. RESULTS: We reviewed 140 cases. The average age of patients was 52.3 years. Eighty tumors were serous, 47 mucinous, 11 mixed, and two endometrioid. The mean diameter overall was 13.7 cm (range 1-70 cm), 10.2 cm for serous, and 20.1 cm for mucinous. Diagnoses of borderline tumors by frozen and permanent pathology were consistent in 60% of cases. Frozen section interpreted a benign lesion as malignant (overdiagnosed) in 10.7% of cases, and interpreted a malignant lesion as benign (underdiagnosed) in 29.3%. No variable was a significant predicator of overdiagnosis. In univariate analysis, underdiagnosis was more likely for other types of tumors than serous (P <.001), tumors larger than 20 cm (P =.039), and tumors confined to the ovaries (P =. 009). When all variables were included in a multiple regression model, only histology was a significant predictor of underdiagnosis (P =.039). CONCLUSION: Frozen or permanent pathology reports of diagnoses of borderline tumor were consistent 60% of the time, whereas the positive predictive value of borderline by frozen section was 89.3%. Tumors other than serous are more likely to be misinterpreted. PMID- 10831978 TI - Prognostic nutritional index in relation to hospital stay in women with gynecologic cancer. AB - OBJECTIVE: To estimate the prevalence of malnutrition, correlate it with length of hospital stay, and evaluate laboratory tools to define it in gynecologic oncology. METHODS: Sixty-seven consecutive hospitalized gynecologic oncology patients were evaluated prospectively using the standardized Prognostic Nutritional Index method, based on serum albumin, transferrin, triceps skin fold and skin sensitivity tests, which defines criteria for malnourished and nourished patients. It was correlated with length of hospital stay. The Mann-Whitney test and Pearson's correlation coefficient were used to evaluate statistical relationships. RESULTS: Cancer distribution among study subjects was 39 cervical (58%), 16 uterine (24%), 11 ovarian (16%), and one vulvar (2%). Malnutrition was found in 36 of 67 women (54%; 95% confidence interval [CI] 41%, 66%). The median (interquartile range) hospital stays of nourished women (n = 31) and malnourished women (n = 36) were 6 (range 4-7) days and 8 (range 6-16) days, respectively (two sided P =.004). That difference remained after controlling for age, extent of metastases, and cancer sites. Albumin correlated well with Prognostic Nutritional Index (R = -.78; 95% CI -.86, -.66; P <.001). Albumin also correlated with length of hospital stay R = -.41; 95% CI -.56, -.25; P <.001). CONCLUSION: Malnutrition is common in gynecologic oncology patients and contributes to longer hospital stays. Albumin is a good substitute for the Prognostic Nutritional Index laboratory test for assessing malnutrition. PMID- 10831979 TI - Routine hematocrit after elective gynecologic surgery. AB - OBJECTIVE: To determine the clinical use and associated costs of routine postoperative hematocrit after elective gynecologic surgery. METHODS: We reviewed the charts of all women who had elective gynecologic surgery over 12 months at a community hospital. Demographic data, estimated blood loss at surgery, pre- and postoperative hematocrits, postoperative symptoms suggesting anemia, and incidence of transfusion were tabulated. Laboratory charges for hematocrit at the hospital were used to develop an economic assessment. Statistical analysis was done with Student t test, Mann-Whitney U test, and Fisher exact test. RESULTS: Among 1105 women who had elective surgery, a single postoperative hematocrit was done in 761 (69%) and two or more postoperative hematocrits (mean 2.4, range 2-6) were done in 201 (18%). The overall incidence of blood transfusion related to elective gynecologic surgery was 1.9% (21 of 1105). Five of those women needed preoperative transfusions, eight needed intraoperative transfusions, and the remaining eight had postoperative transfusions. All women who needed postoperative transfusion showed some sign or symptom of anemia. None received transfusions based solely on hematocrit. Risk factors for postoperative transfusion included low preoperative hematocrit and increased intraoperative blood loss. CONCLUSION: Routine postoperative hematocrit in asymptomatic women after uncomplicated elective gynecologic surgery did not improve outcome. PMID- 10831980 TI - Matrix metalloproteinase-9 and tensile strength of fetal membranes in uncomplicated labor. AB - OBJECTIVE: To analyze the relation between tensile strength and levels of matrix metalloproteinase-9, tissue inhibitor of metalloproteinase-1, and tissue inhibitor of metalloproteinase-2 at a number of sites in human fetal membranes. METHODS: Tensile strengths of fetal membranes from five women who delivered vaginally at term were measured by the method of modified force application. A piece of membrane at each measured site was then dissected, and the levels of matrix metalloproteinase-9, tissue inhibitor of metalloproteinase-1, and tissue inhibitor of metalloproteinase-2 were measured by sandwich enzyme immunoassay. The relationship between tensile strength and enzyme levels was evaluated by Scheffe F test at a total of 81 sites on the five membranes. RESULTS: The mean tensile strength of the membranes was 45.3 +/- 19.8 (mean +/- standard deviation) mmHg/0.3 mm(2) (n = 81). When the measured sites were divided according to tensile strength into four groups (<25, 25-49, 50-74, and >/=75 mmHg/0.3 mm(2)), the level of matrix metalloproteinase-9 (0.72 +/- 0.82 nmol/g protein, n = 12) in the less than 25 mmHg/0.3 mm(2) group was significantly higher than the other groups (0.35 +/- 0.22, 0.28 +/- 0.15, and 0.15 +/- 0.08 nmol/g protein; n = 39, 23, and 7, respectively). The significance level was still higher when the molar ratio of matrix metalloproteinase-9 to tissue inhibitor of metalloproteinase-1 was used for comparison. CONCLUSION: An increased molar ratio of matrix metalloproteinase-9 to tissue inhibitor of metalloproteinase-1 might be related to decreased tensile strength of human fetal membranes in uncomplicated labor. PMID- 10831981 TI - Validity of ultrasound estimation of fetal weight. AB - OBJECTIVE: To assess the validity of ultrasound estimation of fetal weight at term. METHODS: We conducted a prospective observational study whereby all ultrasonic biometric measurements were done by a single observer. Fifty pregnant women at term had ultrasonic measurement of various fetal biometric parameters performed within a week of delivery. Fetal weight was estimated by the use of four reported methods (Aoki, Campbell, Shepard, and Hadlock formulas). We compared estimated weight with the birth weight after the estimated fetal weight was adjusted by adding 25 g for each day between the ultrasound measurements and delivery. RESULTS: The adjusted estimated fetal weight obtained from all four formulas tended to be lower than measured birth weight. The smallest mean difference was obtained with the Shepard and Aoki formulas (51.4 g and 60.5 g, respectively), whereas the Campbell and Hadlock formulas produced larger mean differences (141.8 g and 190.7 g, respectively). The Aoki formula generated the smallest range between the limits of agreement (-324.2 to 445.2 g) whereas the Campbell formula produced the largest range (-286.5 to 570.1 g). The range between the limits of agreement generated with the Shepard and Hadlock formulas were intermediate between those produced by the Aoki and Campbell formulas. The intraclass correlation coefficients generated with the Aoki and Shepard formulas were identical (0.90). The intraclass correlation coefficients obtained with the Hadlock (0.84) and Campbell formulas (0.85) were lower. CONCLUSION: The validity of ultrasonic estimation of fetal weight at term with all four formulas was high. PMID- 10831982 TI - Chorioamnionitis and uterine function. AB - OBJECTIVE: To clarify the relationships between intrapartum chorioamnionitis and abnormalities of uterine function, including labor abnormalities, cesarean delivery, and hemorrhage during parturition. METHODS: We did a retrospective cohort study on 16,226 deliveries between 1986 and 1996 that were identified from the University of California, San Francisco Perinatal Database. Variables included maternal age, parity, birth weight, gestational age, epidural usage, oxytocin usage, cesarean delivery, labor abnormalities, length of labor stages, estimated blood loss, and chorioamnionitis status. Analyses included chi(2), Student t test, and logistic regression. RESULTS: In the chorioamnionitis group, there was a statistically significantly higher proportion of women with labor abnormalities (relative risks [RRs] 2.6-4.1), cesarean delivery (RR 3.3; 95% confidence interval [CI] 3.0, 3.5), hemorrhage after cesarean (RR 1.2; 95% CI 1.1, 1.3), and hemorrhage after vaginal delivery (RR 2.1; 95% CI 1.9, 2.4) compared with the nonchorioamnionitis group. Those findings stayed significant after multivariate analysis for various labor abnormalities (adjusted odds ratios [ORs] ranged 1.3-2.1), cesarean delivery (adjusted OR 1.8; 95% CI 1.5, 2.1), hemorrhage after cesarean (adjusted OR 1.5; 95% CI 1.2, 1.8), and hemorrhage after vaginal delivery (adjusted OR 1.8; 95% CI 1.5, 2.1). CONCLUSION: Adverse associations between chorioamnionitis and labor abnormalities, cesarean rate, and hemorrhage during parturition appear to be significant, suggesting a deleterious effect of chorioamnionitis on uterine function. PMID- 10831983 TI - Increased adverse pregnancy outcomes with unreliable last menstruation. AB - OBJECTIVE: To estimate the risk of adverse outcomes in women whose first day of the last menstrual period (LMP) was unreliable. METHODS: Among 20,244 singleton pregnancies with measurements of biparietal diameter between 12 and 22 weeks' gestation, LMP was registered as unreliable in 3775 (18.6%) and reliable in 16,469 (81.4%). Adverse outcomes were defined as spontaneous or missed abortions after 12 weeks' gestation, stillbirth or postnatal death within 1 year, preterm birth, birth weight less than 2500 g, and low birth weight (LBW) for gestation (lower than 22% below sex-specific expected weight). Logistic regression analysis and Kaplan-Meier survival analysis were used to analyze the risk of adverse outcomes. RESULTS: The risk of death was doubled in pregnant women with unreliable LMPs compared with those with reliable LMPs (odds ratio [OR] 2.0; 95% confidence interval [CI] 1.5, 2.6). This risk was highest with respect to stillbirth (OR 2.7; 95% CI 1.7, 4.3). The risks of preterm birth, LBW, and LBW for gestation were also significantly increased (ORs 1.5, 1.4, and 1.2; 95% CIs 1.3, 1.7; 1.2, 1.6; and 1. 0, 1.4, respectively). CONCLUSION: An unreliable LMP is associated with increased risk of adverse outcomes, especially fetal death. PMID- 10831984 TI - Barriers to timely prenatal care among women with insurance: the importance of prepregnancy factors. AB - OBJECTIVE: This study, designed to avoid methodologic limitations of previous research, aimed to identify the important noninsurance barriers to timely prenatal care. METHODS: We identified a subsample of a cross-sectional statewide representative postpartum survey conducted in California during 1994-1995, focusing on 3071 low-income women with Medi-Cal or private coverage throughout pregnancy. RESULTS: Twenty-eight percent of those women had untimely care, although only 6% were unaware of their pregnancies during the first trimester. Controlling for numerous sociodemographic factors; knowledge, attitudes, beliefs, and behaviors; stressful life circumstances; and logistic obstacles that might deter seeking or receiving care, the following risk factors for untimely care were significant and experienced by more than one fifth of women: unwanted or unplanned pregnancy (affecting 43% and 66% of women, respectively), no regular provider before pregnancy (affecting 22% of women), and no schooling beyond high school (affecting 76% of women). Transportation problems, affecting 8% of women, appeared to be the only significant logistic barrier to timely care. CONCLUSION: Improving timeliness of prenatal care among low-income women with third-party coverage is likely to require broad social and health policies that focus on factors affecting women before pregnancy. Assistance with transportation could contribute to more timely care for some low-income women, but programs focusing primarily on other noninsurance barriers during pregnancy might not substantially improve the timeliness of care, at least among low-income women with third-party coverage. PMID- 10831985 TI - Prenatal diagnosis of congenital cytomegalovirus infection: prospective study of 237 pregnancies at risk. AB - OBJECTIVE: To develop recommendations for prenatal diagnosis of congenital cytomegalovirus (CMV) infection and evaluate possible prognostic markers. METHODS: We studied 237 pregnant women who had suspected or confirmed primary CMV infections by amniocenteses with or without funipuncture. Diagnosis of CMV was based on culture and polymerase chain reaction (PCR) done on amniotic fluid (AF) samples; fetal blood tests for CMV immunoglobulin M antibodies, PCR, and nonspecific biologic markers; and repeated ultrasound examinations. In cases of pregnancy termination, viral and pathologic examinations of fetuses were done. At birth, CMV infections were sought in newborns. Pediatric follow-up was scheduled for at least 2 years. RESULTS: Of 210 fetuses and newborns correctly evaluated, 55 had CMV infections. Ten of 38 fetuses infected before 20 weeks' pregnancy had severe congenital disease. The global sensitivity of prenatal diagnosis was 80%. Best sensitivity and 100% specificity were achieved by PCR done on AF sampled after 21 weeks' gestation, respecting a mean interval of 7 weeks between diagnosis of maternal infection and prenatal diagnosis. Fetal thrombocytopenia was associated with severe fetal disease. Ultrasound follow-up missed two fetuses who presented with neurologic impairment due to CMV after birth. CONCLUSION: A reliable prenatal diagnosis of congenital CMV infection based on PCR on amniocentesis samples can be made after 21 weeks' pregnancy, after a 7-week interval between diagnosis of maternal infection and antenatal procedure. Ultrasound and nonspecific biologic parameters are not sufficient to identify all fetuses at risk of severe sequelae. PMID- 10831986 TI - Urine hyperglycosylated hCG plus ultrasound biometry for detection of down syndrome in the second trimester in a high-risk population. AB - OBJECTIVE: To evaluate measurement of levels of urine hyperglycosylated hCG, a form of hCG with abnormally branched oligosaccharide side chains, in conjunction with ultrasound biometry for Down syndrome risk prediction in an at-risk group. METHOD: We prospectively measured urine hyperglycosylated hCG levels, humeral length, and nuchal thickness in women who had second-trimester amniocentesis. Urine hyperglycosylated hCG levels were measured by a two-step enzyme immunometric assay using monoclonal antibody beta152. Humeral length, nuchal thickness, and hyperglycosylated hCG values were expressed as multiples of the median, and the Down syndrome screening efficiency of the three analytes plus age was determined. A receiver operating characteristic (ROC) curve was generated, and the area under the curve was used to assess the Down syndrome screening performance of the algorithm. RESULTS: There were 23 cases of Down syndrome among 1016 singleton pregnancies. Mean gestational age (+/- standard deviation) was 16.1 +/- 1.2 weeks at the time of amniocentesis. Mean maternal age was 37.1 +/- 3.2 years. Biometry and measurement of hyperglycosylated hCG levels had a 91.3% detection rate at a 3.2% false-positive rate and a 100% detection rate at a 10.7% false-positive rate. The area under the ROC curve was 0.986 (P <.001), and that for measurement of hyperglycosylated hCG levels plus age was 0.941 (P <.001). The area under the curve was significantly larger with combined biochemical and biometry markers compared with measurement of hyperglycosylated hCG levels plus age alone (P <.02), proving that the former was superior to the latter. CONCLUSION: A new Down syndrome biochemical marker combined with ultrasound biometry had a high screening efficiency in a high-risk group. All cases of Down syndrome in this study population would have been detected at an amniocentesis rate of less than 10.7%. Our results appear superior to those found with other second-trimester algorithms. The combination is promising as an alternative to "automatic" genetic amniocentesis in women of advanced maternal age and other high-risk groups. PMID- 10831987 TI - Knowledge and clinical practice regarding folic acid among obstetrician gynecologists. AB - OBJECTIVE: To assess obstetrician-gynecologists' knowledge of and clinical practice concerning folate. METHODS: We mailed surveys on nutrition during pregnancy to the 230 ACOG Fellows who are members of the Collaborative Ambulatory Research Network and to a random sample of 800 Fellows who are not members of the Network. Our results focus on questions concerning folate. RESULTS: We analyzed 488 surveys (a 47.4% response rate). Approximately two thirds of respondents screen their pregnant patients for folate intake. Fewer (53%) screen their nonpregnant patients of childbearing age. Those who screened their patients for folate intake were more likely to counsel pregnant patients about diet. They also were more likely to believe that nutritional counseling would improve pregnancy outcomes (70.0% versus 56.5%) and overall patient health (77.5% versus 66.5%). Most Fellows were aware that macrocytic anemia was a manifestation of folate deficiency (90.4%) and that folic acid supplementation during preconception and the early prenatal period helps protect against neural tube defects (96.5%). They were aware that alcoholics (91.4%), smokers (61.3%), and lactating women (53.5%) are at increased risk of folate deficiency. They were less aware of other consequences of low folate intake, such as increased serum homocysteine (20.3%). Respondents who screen their pregnant patients for folate intake correctly answered more of the knowledge questions about folate than physicians who do not screen. CONCLUSION: Obstetrician-gynecologists are generally aware of the link between folate intake and neural tube defects, but are less aware of other aspects of folate metabolism. PMID- 10831988 TI - Maternal morbidity and mortality associated with multiple gestations. AB - OBJECTIVE: To test the hypothesis that women with multiple gestations are at increased risk of adverse maternal outcomes. METHODS: We studied the association between multiple gestation and frequency of adverse maternal outcomes in 885,338 pregnancies recorded in the Perinatal Information System database of the Latin American Center for Perinatology and Human Development, Montevideo, Uruguay, between 1985 and 1997. Relative risks (RRs) were adjusted for 14 potential confounding factors through multiple logistic regression models. RESULTS: There were 15,484 multiple gestations. Among parous women, multiple gestation was associated with a twofold increase in risk of death compared with singleton gestations [adjusted RR 2.1; 95% confidence interval (CI) 1.1, 3.9]. Compared with singleton gestations, women with multiple gestations had adjusted RRs of 3.0 (95% CI, 2.9, 3.3) for eclampsia, 2.2 (95% CI, 1. 9, 2.5) for preeclampsia, and 2.0 (95% CI, 1.9, 2.0) for postpartum hemorrhage. Likewise, there was significant association between multiple gestation and increased incidence of preterm labor, anemia, urinary tract infection, puerperal endometritis, and cesarean delivery. The incidences of premature rupture of membranes, third-trimester bleeding, and gestational diabetes mellitus were not statistically different for singleton and multiple gestations. CONCLUSION: Multiple gestation increases the risk of significant maternal morbidity and mortality. PMID- 10831989 TI - A randomized comparison of oral and intravaginal misoprostol for labor induction. AB - OBJECTIVE: To compare oral misoprostol (100 microgram) to vaginal misoprostol (25 microgram) for cervical ripening and labor induction. METHODS: Two hundred thirty six women with medical or obstetric indications for labor induction and undilated, uneffaced cervices were assigned randomly to receive 100 microgram of oral or 25 microgram of vaginal misoprostol every 4 hours for 24 hours. Intravenous oxytocin was then given using a standardized protocol. We anticipated that 31% of women treated orally and 50% of women treated vaginally would deliver vaginally within 24 hours of starting labor induction. RESULTS: Among 234 subjects, 121 received oral and 113 received vaginal misoprostol. The mean (+/- standard deviation [SD]) interval from start of induction to delivery was 1240 +/ 845 minutes for orally treated women and 1381 +/- 802 minutes for vaginally treated women (P =.06, log-transformed data). More orally treated women delivered vaginally in 24 hours than vaginally treated women (74 versus 54, P =.14, relative risk [RR] 1.14, 95% confidence interval [CI] 0.92, 1.40). One hundred six women (87.6%) who received oral misoprostol delivered vaginally, compared with 88 women (77.9%) who received vaginal misoprostol (P =.07). Oxytocin was given to 60 (49. 6%) orally treated and 59 (52.2%) vaginally treated subjects (P =. 69). More women in the oral group had tachysystole, nine compared with two (P =.06), hypertonus, and hyperstimulation. Frequencies of intrapartum complications and birth outcomes were similar between groups. CONCLUSION: Oral misoprostol 100 microgram and vaginal misoprostol 25 microgram were similarly effective for cervical ripening and labor induction. Oral administration was associated with trends toward higher likelihood of vaginal delivery and more uterine tachysystole. PMID- 10831990 TI - Chorioamnionitis and uterine function AB - Objective: To clarify the relationships between intrapartum chorioamnionitis and abnormalities of uterine function, including labor abnormalities, cesarean delivery, and hemorrhage during parturition.Methods: We did a retrospective cohort study on 16,226 deliveries between 1986 and 1996 that were identified from the University of California, San Francisco Perinatal Database. Variables included maternal age, parity, birth weight, gestational age, epidural usage, oxytocin usage, cesarean delivery, labor abnormalities, length of labor stages, estimated blood loss, and chorioamnionitis status. Analyses included chi(2), Student t test, and logistic regression.Results: In the chorioamnionitis group, there was a statistically significantly higher proportion of women with labor abnormalities (relative risks [RRs] 2.6-4.1), cesarean delivery (RR 3.3; 95% confidence interval [CI] 3.0, 3.5), hemorrhage after cesarean (RR 1.2; 95% CI 1.1, 1.3), and hemorrhage after vaginal delivery (RR 2.1; 95% CI 1.9, 2.4) compared with the nonchorioamnionitis group. Those findings stayed significant after multivariate analysis for various labor abnormalities (adjusted odds ratios [ORs] ranged 1.3-2.1), cesarean delivery (adjusted OR 1.8; 95% CI 1.5, 2.1), hemorrhage after cesarean (adjusted OR 1.5; 95% CI 1.2, 1.8), and hemorrhage after vaginal delivery (adjusted OR 1.8; 95% CI 1.5, 2.1).Conclusion: Adverse associations between chorioamnionitis and labor abnormalities, cesarean rate, and hemorrhage during parturition appear to be significant, suggesting a deleterious effect of chorioamnionitis on uterine function. PMID- 10831991 TI - Labor after previous cesarean: influence of prior indication and parity. AB - OBJECTIVE: To determine whether the risk of cesarean for women who had trials of labor after one prior cesarean differs from that of nulliparas overall and by indications for those cesareans. METHODS: We reviewed medical records of women who had trials of labor after cesareans between July 1984 and June 1996, and of nulliparas who delivered between December 1994 and August 1995. Cesarean rates for women with prior cesareans were compared with the rates for nulliparas overall and by prior cesarean indication (breech, failure to progress, nonreassuring fetal testing, or other). Lengths of labor for women who had repeat cesareans for failure to progress in index pregnancies were compared by prior cesarean indication. RESULTS: The cesarean rate was 28.7% (634 of 2207) for the prior cesarean group and 13.5% (219 of 1617) for nulliparas (P =.001), and varied according to the prior cesarean indication (13.9%, 37.3%, 25. 4%, and 24.8% for breech, failure to progress, nonreassuring fetal testing, and other, respectively). Mean durations of labor in the index pregnancies for women who had cesareans for failure to progress were 13.9, 11.5, 13.4, and 15.1 hours for breech, failure to progress, nonreassuring fetal testing, and other, respectively. CONCLUSION: Overall rates of cesareans were higher for women with one prior cesarean than for nulliparas. Rates of cesareans after trials of labor were related to the prior cesarean indications. Rates were highest for women whose prior cesareans were for failure to progress and lowest for women whose prior cesareans were for breech. The latter group had a rate that was essentially identical to that of nulliparas. Among women with cesareans for failure to progress in index pregnancies, lengths of labor were shorter for those whose prior cesareans were for failure to progress than for those whose prior cesareans were for other indications, suggesting that physicians may intervene earlier in these cases. PMID- 10831992 TI - Elective induction of labor as a risk factor for cesarean delivery among low-risk women at term. AB - OBJECTIVE: To determine the effects of elective induction on the risk of cesarean delivery in a cohort of women with low-risk term pregnancies and to evaluate the costs of elective induction services within our hospital system. METHODS: Records of 1135 eligible women with low-risk, singleton, vertex pregnancies at 38-41 weeks' gestation who were eligible for vaginal delivery were analyzed retrospectively after elective induction (n = 263) or spontaneous labor (n = 872). Outcome measures included cesarean delivery and direct costs. Variables evaluated were parity, maternal age, estimated gestational age, birth weight, prior cesarean delivery, epidural anesthetic use, and provider category. Analysis was by univariable and multivariable regression modeling. RESULTS: Elective induction placed nulliparas at a twofold higher risk for cesarean delivery (odds ratio 2.4, 95% confidence interval 1.2, 4.9) after adjustment for birth weight, maternal age, and gestational age. We found a significantly increased risk of cesarean delivery with increased birth weight for nulliparas (2-66.7%). Increasing maternal age increased the risk of cesarean delivery in all parity groups (P <.05), but particularly among nulliparas (3-26.3%) (P <.001). Electively induced labors that ended in vaginal delivery cost $273 more and required an average of 4 hours more in the hospital before delivery than did noninduced vaginal deliveries (P <.001). CONCLUSION: Elective induction significantly increased the risk of cesarean delivery for nulliparas, and increased in-hospital predelivery time and costs. PMID- 10831993 TI - Wound infection after cesarean: effect of subcutaneous tissue thickness. AB - OBJECTIVE: To estimate the effect of the thickness of subcutaneous tissue at the surgery site on abdominal wound infection after cesarean delivery. METHODS: We measured the maximum vertical depths of subcutaneous incisions of women who had cesarean deliveries. The surgical technique for closure was standardized and drains were not used. Abdominal wound infection was defined by standard criteria and limited to the first 6 postoperative weeks. Additional demographic, intrapartum, and perioperative data previously associated with wound infection also were collected. Data were analyzed by Student t test, chi(2) test, and multiple logistic regression. RESULTS: Wound infection occurred in 11 of 140 women (7.8%) who delivered by cesarean. Risk factors identified as significantly associated with wound infection by univariate analysis were thickness of subcutaneous tissue, maternal weight, and body mass index. Multiple logistic regression analysis confirmed subcutaneous tissue thickness as the only significant risk factor for wound infection, with a relative risk of 2.8 (95% confidence interval 1.3, 5.9). There were no significant differences between women who developed wound infections and those without infections in terms of selected demographics, duration of ruptured membranes, number of vaginal examinations, chorioamnionitis, type of skin incision, or duration of surgery. CONCLUSION: Thickness of subcutaneous tissue appears to be the only significant risk factor associated with abdominal wound infection after cesarean delivery. PMID- 10831994 TI - Pregnancy after adjustable gastric banding. AB - OBJECTIVE: To determine outcomes of pregnancies of obese women who had surgical placement of an adjustable gastric band to treat obesity. METHODS: We conducted two clinical trials to evaluate adjustable gastric banding that involved 359 obese women of reproductive potential (age 18-51 years), of whom 20 conceived resulting in 23 pregnancies. Specific information about pregnancies and fetal outcomes was collected from medical records and direct patient contact. RESULTS: Eighteen pregnancies were full term, one was an ectopic gestation, two ended in elective abortions, and two in spontaneous abortions unrelated to the womens' medical conditions. Of the 18 full-term pregnancies, 14 delivered vaginally and four by cesarean (one for prolonged fetal bradycardia, two for cephalopelvic disproportion, and one repeat cesarean for twins). The mean birth weight was 3676 g (range 2381-3912 g). Five women lost weight (range 1.8-17.6 kg) during pregnancy without obvious fetal and neonatal effects. Three women had fluid removed from their gastric bands (decreasing the mechanical constriction) to treat nausea and vomiting. Two women who had no fluid in their bands eliminated the effectiveness of the obesity treatment, resulting in excessive weight gain. CONCLUSION: Morbidly obese women who became pregnant soon after receiving an adjustable gastric band had uncomplicated pregnancies. Adjustment of the gastric band to decrease the amount of mechanical obstruction decreased nausea and vomiting, but led to excessive weight gain in two women when it was done prophylactically. Obese women at risk of pregnancy should be counseled that it might occur unexpectedly after weight loss from gastric banding unless birth control is promptly instituted. PMID- 10831995 TI - A survey of pessary use by members of the American urogynecologic society. AB - OBJECTIVE: To describe trends in pessary use for pelvic organ prolapse. METHODS: An anonymous survey administered to the membership of the American Urogynecologic Society covered indications, management, and choice of pessary for specific support defects. RESULTS: The response rate was 48% (359 of 748). Two hundred fifty surveys were received at the scientific meeting and 109 were returned by mail. Seventy-seven percent used pessaries as first-line therapy for prolapse, while 12% reserved pessaries for women who were not surgical candidates. With respect to specific support defects, 89% used a pessary for anterior defects, 60% for posterior defects, 74% for apical defects, and 76% for complete procidentia. Twenty-two percent used the same pessary, usually a ring pessary, for all support defects. In the 78% who tailored the pessary to the defect, support pessaries were more common for anterior (ring) and apical defects (ring), while space filling pessaries were more common for posterior defects (donut) and complete procidentia (Gellhorn). Less than half considered a prior hysterectomy or sexual activity contraindications for a pessary, while 64% considered hypoestrogenism a contraindication. Forty-four percent used a different pessary for women with a prior hysterectomy and 59% for women with a weak pelvic diaphragm. Ninety-two percent of physicians believed that pessaries relieve symptoms associated with pelvic organ prolapse, while 48% felt that pessaries also had therapeutic benefit in addition to relieving symptoms. CONCLUSION: While there are identifiable trends in pessary use, there is no clear consensus regarding the indications for support pessaries compared with space-filling pessaries, or the use of a single pessary for all support defects compared with tailoring the pessary to the specific defect. Randomized clinical trials are needed to define optimal pessary use. PMID- 10831996 TI - Coumarin derivatives and breast-feeding. AB - Coumarin derivatives are the anticoagulants most widely used in the United States. These agents are relatively contraindicated during pregnancy, and the use of these drugs in breast-feeding women remains controversial. Much of the confusion regarding the passage of these agents into breast milk might stem from the fact that different agents possess significantly different chemical properties. A review of the chemical structure of different coumarin derivatives, as well as available clinical evidence, suggests that warfarin sodium is not excreted into breast milk, and can be safely given to women requiring therapeutic anticoagulation postpartum. For the rare patient who cannot tolerate warfarin sodium, the use of dicumarol, rather than anisindione, is preferred. PMID- 10831997 TI - Erb's palsy: concepts of causation. AB - In the past, obstetric textbooks have stated (without evidence) that Erb's palsy is caused by the birth attendant. The mechanism cited is stated to be the application of excessive lateral traction placed on the fetal head and neck. Recent research findings refute this unproven theory. Findings include (1) only 50% of cases are associated with shoulder dystocia; (2) the 4.7-fold increase in the condition was associated with a precipitate second stage compared with one of normal length; and (3) the same injury rate with direct manipulation techniques (for resolving shoulder dystocia) was comparable to that of McRobert's position and/or suprapubic pressure. The overwhelming evidence indicts the propulsive nature of the stretching of the involved nerves over which the birth attendant has no control. PMID- 10831998 TI - Detection and management of eating disorders during pregnancy. AB - Eating disorders such as anorexia nervosa and bulimia nervosa are associated with potential negative consequences during pregnancy, including higher rates of miscarriage, low birth weight, obstetric complications, and postpartum depression. Women with eating disorders are reluctant to disclose symptoms to health care providers, so it is important for obstetric clinicians to be aware of warning signs and assessment techniques to identify them. Signs suggestive of eating disorders include lack of weight gain, hyperemesis gravidarum, and a history of eating disorders. Recent studies showed that the Eating Disorder Examination, a newly developed assessment tool, can ascertain whether someone has an eating disorder. Questions from the Eating Disorder Examination about body image, food avoidance, food rules, and dieting behaviors have successfully differentiated women with eating disorders from healthy controls. We offer an approach to the clinical care of individuals identified or suspected of having eating disorders. We recommend a team approach that emphasizes ongoing communication and clear goal setting for the care of pregnant women with eating disorders. PMID- 10831999 TI - Heart rate variability, trait anxiety, and perceived stress among physically fit men and women. AB - BACKGROUND: It is unclear from prior reports whether the relationships between self-ratings of anxiety or emotional stress and parasympathetic nervous system components of heart rate variability are independent of personality and cardiorespiratory fitness. We examined those relationships in a clinical setting prior to a standardized exercise test. METHODS AND RESULTS: Heart rate variability (HRV) was measured during 5 min of supine rest among 92 healthy men (N=52) and women (N=40) who had above-average cardiorespiratory fitness as indicated by peak oxygen uptake measured during grade-incremented treadmill exercise. HRV datasets were decomposed into low-frequency (LF; 0.05-0.15 Hz) and high-frequency (HF; 0.15-0.5 Hz) components using spectral analysis. Self-ratings of trait anxiety and perceived emotional stress during the past week were also assessed. CONCLUSIONS: There was an inverse relationship between perceived emotional stress during the past week and the normalized HF component of HRV (P=0.038). This indicates a lower cardiac vagal component of HRV among men and women who perceived more stress. That relationship was independent of age, gender, trait anxiety, and cardiorespiratory fitness. It was also independent of heart rate; mean arterial blood pressure; and respiration rate, factors which can influence HRV and might be elevated among people reporting anxiety and perceived stress. We conclude that vagal modulation of heart period appears to be sensitive to the recent experience of persistent emotional stress, regardless of a person's level of physical fitness and disposition toward experiencing anxiety. PMID- 10832000 TI - Top-down modulation of early selective attention processes in children. AB - It has been demonstrated in adults that attention can modulate very early stages of perceptual/cognitive processing, but it has not been determined if this capacity for modulation develops with age. We investigated early attentional processes in parallel and serial visual search tasks in 40 children 7-12 years old, using event-related potentials (ERPs). Two single-feature pop-out tasks were used to assess parallel processing; a conjunction of features task was used to study serial processing. There were significant decreases in latencies with age for the peaks measured [posterior P1, N1 and anterior P2 (P2a)]. P1 and N1 latency effects also varied with trial type and were consistent with top-down modulation of processing, which differed between the parallel tasks. P2a amplitude for the hit trials decreased with increasing age for the colour and serial tasks. Increasing R>L hemispheric asymmetries with age in P2a amplitude were seen on non-target trials for the size and serial tasks only, reflecting serial processing. These data demonstrate that developmental changes in visual selective attention for early stages of processing are continuing through childhood, and that there is top-down modulation at these latencies in children. PMID- 10832001 TI - P300 habituation from auditory single-stimulus and oddball paradigms. AB - The habituation of auditory P300 event-related brain potential (ERP) from single stimulus paradigm was evaluated and compared to that from oddball paradigm. Three task conditions were: oddball with a button press (oddball/press) response, single-stimulus with a button-press (single-stimulus/press) response and a silent count (single-stimulus/count) response. The oddball/press condition demonstrated larger P300 amplitude and longer latency overall than either single-stimulus condition, but P300 amplitude decreased and peak latency increased similarly over successive trial blocks for all three tasks. Thus, the oddball and single stimulus ERP tasks produce analogous changes under repeated measurements and indicate that the single-stimulus task can serve as an alternative method for eliciting the P300 in applied and clinical settings. PMID- 10832002 TI - Changes in EEG alpha power during simulated driving: a demonstration. AB - The aim was to assess the suitability of EEG-based techniques to recording activity during a driving simulation task. To achieve this, an inexpensive driving simulator (comprising a steering wheel, pedals and gear shift) were made to function with a personal computer running 'Need for Speed' simulation software. Simulators of this type are both inexpensive and relatively realistic. The EEG was recorded from four sites on the scalp (P3, P4, F3, F4) for two laps during the driving task, and during a replay task. The driving task involved participants driving a vehicle on a simulated undulating, sealed surface circuit, without any other vehicles present. Two men were participants in this experiment. Power spectra were computed and integrated to produce values of relative alpha activity for each channel and recording epoch, a time-series of alpha activity during each recorded segment. Overall values for alpha activity indicated an increase for replay compared to driving, and also driving on lap 5 compared to driving on lap 2. The EEG changes are consistent with the notion of overall reduction of attention during the later laps and the replay task and indicate the potential of such measures for complex motor behaviour. PMID- 10832003 TI - Delayed verbal reactions are specifically impaired in patients with schizophrenia. AB - Recent neuropsychological, psychophysiological and neuroimaging studies have shown that specific changes in attention and information processes occur in schizophrenic disorders. A verbal delayed reaction methodology, determined by our group since 1992, has provided similar quantitative data. Visual verbal stimuli, occurring in random sequence, are presented to the subject who is requested to utter immediately the perceived word in a first trial; in a second one, a delay (foreperiod) between the word presentation and a 'go' signal (eight asterisks) is interposed. Acousticograms and orbicularis oris EMG are recorded as responses. The ratio of the 1.5-s foreperiod delayed reaction time to the immediate reaction time reveals a facilitation of the executive system occurring during sustained purposeful attention, a characteristic function of the prefrontal cortex. A further ratio, with a 0.1-s foreperiod, has been measured to investigate the effect of interference processes. These trials have been performed including 18 patients with schizophrenia, 31 with idiopathic Parkinson's disease, four patients with extrapyramidal degenerative diseases, five affected by obsessive compulsive disorder and in age-matched normal controls. Both ratios were significantly increased in schizophrenic patients and, on the contrary, decreased in patients with obsessive-compulsive disorder. These changes are in agreement with the neuroimaging findings of 'hypofrontality' in the schizophrenic patients. Verbal delayed reaction methodology seems a reliable and easily applicable tool for investigating sustained purposeful attention processes in clinical conditions. PMID- 10832004 TI - Peripheral and central nervous changes in patients with rheumatoid arthritis in response to repetitive painful stimulation. AB - It has been observed that patients with rheumatoid arthritis (RA) respond differently to repetitive painful stimulation. The present study investigated whether this is related to the peripheral or central nervous nociceptive system. EEG-derived potentials and the negative mucosal potential (NMP) from the respiratory epithelium were recorded in response to painful intranasal stimulation with gaseous CO(2). Differences between groups (12 RA patients, 12 controls) were found when stimuli were presented at short intervals. While the NMP did not differ between groups, patients had larger cortical responses to the first stimuli of a series of painful stimuli. This may indicate that in RA central nervous changes of nociceptive processing are present. PMID- 10832005 TI - The relationships of resting baroreflex sensitivity, heart rate variability and measures of impulse control in children and adolescents. AB - The objectives of the present study were to assess: (1) the feasibility of using a non-invasive method to measure baroreflex sensitivity in children and adolescents; (2) the relationships of resting baroreflex sensitivity with resting levels of other cardiovascular variables; and (3) whether baroreflex sensitivity and heart rate variability, two indices of cardiac vagal control at rest, were related to measures of impulse control. Ninety-one Black and White children (ages 8-10) and adolescents (ages 15-17), both female and male, participated in the study. Baroreflex sensitivity, auscultatory blood pressure, EKG-derived heart rate, and the mean successive difference of interbeat intervals were collected during a 10-min rest period. Measures of impulse control came from the Interview for Antisocial Behavior. Baroreflex sensitivity was strongly positively correlated with mean successive difference and negatively correlated with heart rate for all participant groups; baroreflex sensitivity was negatively correlated with diastolic blood pressure, but only for children, males, and Blacks. Increased problems with impulse control was associated with decreased cardiac vagal control, but only in males. The usefulness of this technique as a measure of vagal activation is discussed. PMID- 10832006 TI - Chronic stress in pregnant rats: effects on growth rate, anxiety and memory capabilities of the offspring. AB - Female rats were repeatedly stressed for 10 periods of 15 min by the presence of a cat, at the 10th (S10) or the 19th (S19) gestational day. The litter from stressed females often contained a majority of males or a majority of females, especially in the S19 group. The death of pups was dramatically high in the S19 group and, compared with controls, growth of the surviving animals was slower. When adult, their long-term memory was altered and they exhibited an aversive behavior relative to wide areas. Moreover, cognitive alterations were revealed by the low level of exploration and the inability to rapidly process the relevant environmental cues. These deficits resemble those of psychiatric patients who had been submitted to pre-natal stress. PMID- 10832007 TI - An alternative approach for achieving cardiovascular baseline: viewing an aquatic video. AB - Due to the importance of baseline and recovery levels in the computation of reactivity, two studies were conducted to determine an alternative method to traditional rest for achieving baseline and recovery levels of cardiovascular measurements. Watching a relaxing, aquatic video was compared with a traditional resting baseline to determine the better method for achieving low baseline levels. In addition, watching the video was compared with traditional rest during 5-min post-task recovery periods. Systolic (SBP) and diastolic blood pressure (DBP) decreased more during the baseline period when subjects viewed the video than when subjects rested quietly. Similarly, subjects displayed greater recovery following the mental tasks when they watched a video than when they merely sat quietly. It is recommended that researchers standardize baseline procedures by showing a relaxing video before administering tasks for the assessment of cardiovascular reactivity. PMID- 10832008 TI - Protective effects of huperzine A on beta-amyloid(25-35) induced oxidative injury in rat pheochromocytoma cells. AB - The effects of huperzine A (HupA), a novel acetylcholinesterase inhibitor, on Abeta(25-35)-induced cell lesion, level of lipid peroxidation, antioxidant enzyme activities were investigated in the rat pheochromocytoma line PC12. Following a 48 h exposure of the cells to Abeta(25-35), a significant reduction in cell survival and activities of glutathione peroxidase (GSH-Px) and catalase (CAT), as well as increased production of malondialdehyde (MDA) and superoxide dismutase (SOD) were observed. Preincubation of the cells with HupA prior to Abeta(25-35) exposure elevated the cell survival and GSH-Px and CAT activities, and decreased the level of MDA and SOD activity. The results indicate that HupA has protective effects against Abeta-induced cell toxicity, which might be beneficial for the treatment of Alzheimer's disease. PMID- 10832009 TI - Autonomic dysreflexia increases plasma adrenaline level in the chronic spinal cord-injured rat. AB - Autonomic dysreflexia (AD) occurs in up to 80% of quadriplegics and high paraplegics and is characterized by exaggerated sympathetic reflexes which induced paroxysmal hypertension. The aim of this study was to determine if plasma catecholamine levels increased during autonomic dysreflexia in the chronic spinal cord-injured (SCI) rats with special care to adrenaline. Catecholamine samples were collected before, during and 1 h after AD induced hypertension with colorectal distension. Results showed that plasma noradrenaline and adrenaline levels increased respectively 1.5-fold and 5-fold during AD in the chronic SCI rats. This suggests substantial roles for these two hormones in mediating the cardiovascular changes during AD. Knowledge of catecholamine levels during AD may thus aid in determining pathophysiology and potential pharmacologic treatments of this autonomic dysfunction. PMID- 10832010 TI - Influence of lipid peroxidation on the nicotinic acetylcholine receptors in PC12 cells. AB - The expression of neuronal nicotinic acetylcholine receptors (nAChRs) influenced by lipid peroxidation has been investigated. The results showed that exposure of PC12 cells to a free radical insult (FeSO(4)) induced concentration-dependent increase in level of MDA (malondialdehyde), and a significant decline in MTT [3 (4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide] reduction. In cultured cells exposed to FeSO(4), a significant dose-dependent decrease in [(3)H] epibatidine binding sites was detected and a significant reduction in B(max) value was observed in the high affinity epibatidine binding site. These results demonstrate that lipid peroxidation in cellular membranes can induce a reduction in numbers of neuronal nAChRs. PMID- 10832011 TI - Circadian expression of period 1, period 2, and arylalkylamine N acetyltransferase mRNA in the rat pineal gland under different light conditions. AB - Period 1 (Per1), 2 (Per2) and arylalkylamine N-acetyltransferase (AA-NAT) mRNA levels were determined by semi-quantitative in situ hybridization in rat pineal glands. In agreement with previous reports, AA-NAT mRNA levels were rhythmic in light:dark (LD) cycles and the rhythm persisted in constant dim light (DLL). Per1 and Per2 mRNA also showed significant variations both in LD and DLL. AA-NAT and Per1 mRNA levels showed very similar patterns of variations in LD and DLL to one another, whereas Per2 showed a different pattern of expression from AA-NAT and Per1. Exposure to 30 min of light did not affect the expression of the three genes, while exposure to a longer light pulse (1 or 2 h) decreased AA-NAT and Per1 mRNA levels; Per2 mRNA levels were also decreased but only temporarily. Our results demonstrate that Per1 and Per2 expression in the rat pineal is under circadian control, and suggest Per1 may be regulated by the same mechanism which controls the expression of AA-NAT gene. Per2 seem to be controlled by a different mechanism. PMID- 10832012 TI - Decrease in the nucleo-cytoplasmic volume ratio of rabbit spinal ganglion neurons with age. AB - Nuclear and perikaryal volumes, as well as the mean percentage of perikaryal volume occupied by lipofuscin, were estimated in spinal ganglion neurons of 12, 42 and 79 month-old rabbits. The volume ratio between nucleus and perikaryal cytoplasm (N/C volume ratio) decreased progressively and significantly with increasing age. This was not a consequence of the perikaryal enlargement due to the age-related lipofuscin accumulation since the same result was obtained when the volume occupied by lipofuscin was subtracted from the perikaryal volume. The decrease in the N/C volume ratio may depend on an age-related loss of DNA, a cytoplasmic influence on nuclear size, or other unknown causes. PMID- 10832013 TI - Changes in ventral medullary light reflectance during hypercapnia in awake and sleeping cats. AB - Activity within rostral and intermediate ventral medullary surface areas, measured as 660 nm scattered light changes, was examined in six cats, (five experimental, one control site) following 5% CO(2) challenges during waking, quiet sleep, and rapid eye movement (REM) sleep states. Activity declined to hypercapnia in all states, with a smaller decline in quiet sleep compared to waking, and intermediate values in REM sleep. The decline occurred more rapidly, with a shorter latency, during wakefulness, but with a much slower return to baseline than during quiet sleep. During REM sleep, the latency to nadir and recovery were greater than in other states. Regional patterns of activation emerged which differed in extent of activation between states. PMID- 10832014 TI - Protective effect of etomidate on kainic acid-induced neurotoxicity in rat hippocampus. AB - The goal of this study was to determine whether etomidate has protective effect against kainic acid (KA)-induced neurotoxicity. Administration of etomidate (20 mg/kg i.p.) was performed in sequence, first being just 1 h after KA (10 mg/kg i.p.) injection, then three times at 1-h intervals. Neuronal damages in hippocampus were evaluated by using the acid fuchsin stain to detect cell death and the heat shock protein-70 (HSP-70) induction as an index of cell injury at 24 h after the administration of KA. HSP-70 induction and acid fuchsin positive neurons were increased in CA1 and CA3 regions of hippocampus after KA injection but significantly decreased by etomidate-injection. These results suggest that the etomidate hold potential effect on the protection of neurons against KA induced neurotoxicity. PMID- 10832015 TI - Zinc increases the excitability of dopaminergic neurons in rat substantia nigra. AB - The effect of zinc ions (Zn(2+)) on the neuronal excitability of substantia nigra (SN) where the zinc level is known higher in Parkinsonian brains than that in normal brains has not yet been elucidated. We, therefore, examined the effect of Zn(2+) on the intrinsic electrical properties of dopaminergic SN neurons, using a whole-cell recording method. Zn(2+) hyperpolarized dopaminergic SN neurons at resting state. Also Zn(2+) shortened the duration of evoked spikes, developed a fast afterhyperpolarization, and increased their firing frequency. Voltage-clamp studies showed that Zn(2+) decreased 4-aminopyridine-sensitive outward currents, suggesting that a transient A-like potassium channel be one of the major targets Zn(2+) can modulate in the SN neurons. PMID- 10832016 TI - Development of externally guided grip force modulation in man. AB - We examined the development of externally guided changes of grip force with respect to force rate and direction of force change. Sixty-nine children, 3-6 years of age, and 17 adults produced increasing or decreasing isometric forces on a small cylindrical sensor using a pinch grip with visual feedback. Force changes were instructed with a visual tracking task. Ramp-like paradigms with both force increase and force decrease and two different target force rates were used (0.5 N/s, 1.25 N/s). Precision of force tracking showed clear age effects and was influenced by the required force rates and directions of force change. In adults, tracking errors were much more dependent on target force rate and direction than in children. Up to four years of age, the children tended to overshoot the target force change in a 'jump and wait' manner in all conditions except for fast target force decreases. Older children tended to overshoot only in the condition with slow target force decrease. Adults showed close undershooting in all conditions when following the target. Adults used either a continuous 'following' strategy or a 'see and catch-up' strategy. The distinct effects on tracking errors suggest an age-related change of strategies from a feedforward strategy with intermittent use of sensorimotor feedback towards a fairly parallel and well-integrated feedback and feedforward processing. A critical age appears to be around five to six years. We suggest that these age effects may reflect distinct developmental velocities of neuronal subpopulations of the cortex and of the cerebello-cortical connections. PMID- 10832017 TI - Redox modulation of large conductance calcium-activated potassium channels in CA1 pyramidal neurons from adult rat hippocampus. AB - Redox regulation of BK(Ca) channels was studied in CA1 pyramidal neurons of adult rat hippocampus by using inside-out configuration of patch clamp. Intracellular application of oxidizing agent 5, 5'-dithio-bis(2-nitrobenzoic acid) (DTNB) markedly increased activity of BK(Ca) channels and this stimulating action persisted even after washout. In contrast, the reducing agent dithiothreitol (DTT) had no apparent effects on channel activity but could reverse the pre exposure of DTNB-induced enhancement. The increase in channel activity produced by DTNB was due to shortened closed time as well as prolonged open time. The effects exerted by another redox couple glutathione disulphide and its reducing form were similar as DTNB and DTT. The present results indicate that BK(Ca) channels in CA1 pyramidal neurons can be modulated by intracellular redox potential, and that augmentation of BK(Ca) channels by oxidative stress might contribute to the postischemic electrophysiological alterations of CA1 pyramidal neurons. PMID- 10832018 TI - Immunohistochemical cell types in the terminal nerve ganglion of the cloudy dogfish, Scyliorhinus torazame, with special regard to neuropeptide Y/FMRFamide immunoreactive cells. AB - Previous studies showed immunoreactivities for neuropeptide Y (NPY), molluscan cardioexcitatory tetrapeptide (FMRFamide), and gonadotropin-releasing hormone (GnRH) in the terminal nerve of elasmobranchs. The present immunohistochemical study demonstrated two types of cells, i.e. GnRH- and NPY/FMRFamide-positive cells, in the terminal nerve ganglion of the elasmobranch Scyliorhinus torazame. The second cell type (non-GnRH element) contained a substance with a common structure or epitope recognized by anti-NPY and anti-FMRFamide antibodies. The NPY/FMRFamide-like immunoreactivity was associated with granules 70-130 nm in diameter, found in the cell bodies, axons, and axon endings. PMID- 10832019 TI - Loss of the muscle silent period evoked by transcranial magnetic stimulation of the motor cortex in patients with cervical cord lesions. AB - The silent period following motor evoked potentials in small hand muscles after transcranial magnetic stimulation (TMS) of the human motor cortex is considered to be cortical origin. The authors report three patients with cervical spinal cord lesions who showed loss of the cortical silent period (CSP) after TMS. One patient had traumatic cervical cord injury, and the other two patients had cervical spondylosis. All the patients had cervical cord compression on magnetic resonance imaging. TMS study showed loss of the CSP in both the hand and foot muscles in two patients and only in the foot muscle in one patient. Paired TMS study in one patient with pseudoathetotic hands showed reduced inhibition within the motor cortex. The hand weakness or interrupted sensory afferents might have caused motor cortical reorganization or hyperexcitability, leading to the loss of the CSP. PMID- 10832020 TI - The expanded CAG repeat associated with juvenile Huntington disease shows a common origin of most or all neurons and glia in human cerebrum. AB - We have analyzed the size of the expanded poly(CAG) associated with juvenile Huntington disease in the cerebra and the cerebella of five patients. The expanded poly(CAG) was always longer in the cerebrum than in the cerebellum, but the difference in size varied from patient to patient. Except for one patient who possessed an unusually large expansion, very little heterogeneity of size was detected within the cerebrum or within the cerebellum. The larger size of the expanded poly(CAG) in cerebrum must therefore have resulted from a single expansion event that took place early in cerebral development. In both cerebrum and cerebellum, the size of the expanded allele of gray matter was identical to that of white matter. We conclude that most if not all neurons and glia of cerebrum are descended from a common bipotent precursor, which segregated early in neurogenesis from the lineage leading to cerebellar neurons and glia. PMID- 10832021 TI - Intracerebroventricular infusion of nerve growth factor induces pain-like response in rats. AB - New strategies have recently been developed where infusion of neurotrophic factors into the brain can rescue different neuronal populations. However, negative side effects have been observed in clinical trials infusing nerve growth factor (NGF) into the lateral ventricle in man, namely pain. Little is known about pain behavior in animals after intracerebroventricular (i.c.v.) neurotrophic injections. Thus, we have examined the effects of i.c.v. infusion of NGF for 2 weeks on the behavioral response of rats to mechanical, cold and heat stimulation. Seven micrograms/day of NGF elicited a significant decrease in vocalization threshold to mechanical stimulation and a significantly increased response to cold and heat stimuli as compared with control. The concentration of NGF in cerebrospinal fluid (CSF) was significantly increased as compared with non allodynic rats. The enhanced responses to mechanical and heat, but not to cold, stimulation were significantly reduced by CP-99994, a selective antagonist to tachykinin NK-1 receptors. When NGF was infused into the brain parenchyma (striatum, cortex and septum) no allodynic nor hyperalgesic responses could be detected. These results indicate that in rats i.c.v. but not intraparenchymal infusion of NGF induce mechanical and cold allodynia as well as heat hyperalgesia, which is mediated, at least in part, by activation of NK-1 receptors. PMID- 10832022 TI - No evidence of linkage to 6p markers in spanish families with juvenile myoclonic epilepsy. AB - Juvenile myoclonic epilepsy (JME) is a common subtype of hereditary generalized epilepsy with an uncertain pattern of inheritance. Different studies in multiple families have provided evidence for and against linkage of the disease to chromosome 6p. We performed linkage analysis using microsatellite markers from 6p (D6S109, D6S248, D6S291, D6S426, D6S272, D6S466, D6S294, D6S257) and from centromeric 6q region (D6S402) in seven small families of Spanish origin. The highest LOD scores were obtained under an autosomal dominant inheritance model with a penetrance of 70% but a significant positive LOD score (Z>3) was not reached. LOD scores<-2 were obtained at different markers in three of our families. These results support the concept of genetic heterogeneity in the disease. PMID- 10832023 TI - Cloning of the mouse 25-hydroxyvitamin D(3)-1alpha-hydroxylase (CYP1alpha) gene. AB - A genomic clone for 25-hydroxyvitamin D(3)-1alpha-hydroxylase (1alpha hydroxylase) was isolated from a mouse embryonic stem cell P1 genomic library. It contains nine exons spanning 4.8 kb from the transcriptional start site. All the intron insertion sites are identical to that of the human 1alpha-hydroxylase and human vitamin D(3) 25-hydroxylase genes. A polyadenylation signal AUUAAA that differs from the consensus sequence at the second residue was identified 16 bp downstream of the 3' end of the previously reported mouse cDNA. This element is the only common natural variant described. The 3' end of the gene was determined using a RACE technique. Three poly(A) addition sites were observed 12, 15 and 22 bases from the AUUAAA element. Such distances are in agreement with what is required for maturation of mammalian pre-mRNAs. This molecular cloning makes possible the generation of transgenic mice in order to further investigate the role and importance of the 25-hydroxyvitamin D(3)-1alpha-hydroxylase (CYP1alpha). PMID- 10832024 TI - Chloride-dependent inhibition of vesicular glutamate uptake by alpha-keto acids accumulated in maple syrup urine disease. AB - Maple syrup urine disease is a metabolic disorder caused by mutations of the branched chain keto acid dehydrogenase complex, leading to accumulation of alpha keto acids and their amino acid precursors in the brain. We now report that alpha ketoisovaleric, alpha-keto-beta-methyl-n-valeric and alpha-ketoisocaproic acids accumulated in the disease inhibit glutamate uptake into rat brain synaptic vesicles. The alpha-keto acids did not affect the electrochemical proton gradient across the membrane, suggesting that they interact directly with the vesicular glutamate carrier. Chloride anions have a biphasic effect on glutamate uptake. Low concentrations activate the uptake (0.2 to 8 mM), while higher concentrations are inhibitory. The alpha-keto acids inhibited glutamate uptake by a new mechanism, involving a change in the chloride dependence for the activation of glutamate uptake. The activation of glutamate uptake by low chloride concentrations was shifted toward higher concentrations of the anion in the presence of alpha-keto acids. Inhibition by alpha-keto acids was abolished at high chloride concentrations (20 to 80 mM), indicating that alpha-keto acids specifically change the stimulatory effect of low chloride concentrations. High glutamate concentrations also reduced the inhibition by alpha-keto acids, an effect observed both in the absence and in the presence of low chloride concentrations. The results suggest that in addition to their possible pathophysiological role in maple syrup urine disease, alpha-keto acids are valuable tools to study the mechanism of vesicular transport of glutamate. PMID- 10832025 TI - A silent antifungal protein (AFP)-like gene lacking two introns in the mould Trichoderma viride. AB - In cells of the mould Trichoderma viride, the existence of an antifungal protein (AFP)-like gene consisting of 285 bp was confirmed by Southern analysis that genomic DNA of T. viride could hybridize with the cDNA of mature AFP of Aspergillus giganteus MDH 18894. Except for the absence of two introns, the nucleotide sequence of the AFP-like gene was identical to that of the AFP gene of A. giganteus in positions 336-479, 568-649, and 706-765. The AFP-like gene could not be transcribed into its mRNA in T. viride cells as examined by RT-PCR using total RNAs of T. viride as template. Furthermore, AFP could not be detected either directly from the culture medium of T. viride or by Western analysis. However, the AFP-like gene could be actively expressed like the cDNA of AFP in Escherichia coli cell. Recombinant AFP exhibited similar antifungal activity as native AFP. PMID- 10832026 TI - Liposome-entrapped plasmid DNA: characterisation studies. AB - Plasmid DNA pRc/CMV HBS (5.6 kb) (100 microg) encoding the S (small) region of hepatitis B surface antigen was incorporated by the dehydration-rehydration method into liposomes composed of 16 micromol egg phosphatidylcholine (PC), 8 micromol dioleoylphosphatidylcholine (DOPE) and 1, 2-diodeoyl-3 (trimethylammonium)propane (DOTAP) (cationic liposomes) or phosphatidylglycerol (anionic liposomes) in a variety of molar ratios. The method, entailing mixing of small unilamellar vesicles (SUV) with the DNA, followed by dehydration and rehydration, yielded incorporation values of 95-97 and 48-54% of the DNA used, respectively. Mixing of preformed cationic liposomes with 100 microg plasmid DNA also led to high complexation values of 73-97%. As expected, the association of DNA with preformed anionic liposomes was low (9%). Further work with cationic PC/DOPE/DOTAP liposomes attempted to establish differences in the nature of DNA association with the vesicles after complexation and the constructs generated by the process of dehydration/rehydration. Several lines of evidence obtained from studies on vesicle size and zeta-potential, fluorescent microscopy and gel electrophoresis in the presence of the anion sodium dodecyl sulphate (SDS) indicate that, under the conditions employed, interaction of DNA with preformed cationic SUV as above, or with cationic SUV made of DOPE and DOTAP (1:1 molar ratio; ESCORT Transfection Reagent), leads to the formation of large complexes with externally bound DNA. For instance, such DNA is accessible to and can be dissociated by competing anionic SDS molecules. However, dehydration of the DNA SUV complexes and subsequent rehydration, generates submicron size liposomes incorporating most of the DNA in a fashion that prevents DNA displacement through anion competition. It is suggested that, in this case, DNA is entrapped within the aqueous compartments, in between bilayers, presumably bound to the cationic charges. PMID- 10832027 TI - Characterization of chloroquine-hematin mu-oxo dimer binding by isothermal titration calorimetry. AB - Numerous studies indicate that a key feature of chloroquine's (CQ) antimalarial activity is its interaction with hematin. We now characterize this CQ-hematin interaction in detail using isothermal titration calorimetry (ITC). Between pH 5.6 and 9.0, association constants (K(a) values) for enthalpy-driven CQ-hematin mu-oxo dimer binding fell in the narrow range of 2.3-4.4 x 10(5) M(-1). It is therefore probable that CQ-hematin mu-oxo dimer binding affinity does not diminish at the pH range (4.8-5.4) of the parasite food vacuole. The binding affinity was unaffected by high salt concentrations, suggesting that ionic interactions do not contribute significantly to this complexation. With increasing ionic strength, the entropic penalty of CQ-hematin mu-oxo dimer binding decreased accompanied by increased hematin mu-oxo dimer aggregation. A stoichiometry (n) of 1:4 in the pH range 6.5-9.0 indicates that CQ binds to two hematin mu-oxo dimers. At pH 5.6, a stoichiometry of 1:8 suggests that CQ binds to an aggregate of four hematin mu-oxo dimers. This work adds further evidence supporting the hypothesis that CQ impedes hematin monomer incorporation into hemozoin by producing a forward shift in the hematin monomer-hematin mu-oxo dimer equilibrium, contributing to a destructive accumulation of soluble forms of hematin in the parasite and leading to its death by hematin poisoning. PMID- 10832028 TI - Interaction of antibodies and antigens conjugated with synthetic polyanions: on the way of creating an artificial chaperone. AB - Recently we have initiated the use of synthetic polyelectrolytes to mimic the action of chaperones in living cells [Dainiak et al., Biochim. Biophys. Acta 1381 (1998) 279-285]. The next step in this direction is done by the synthesis of conjugates of poly(methacrylic acid) (PMAA) with antigen, denatured glyceraldehyde-3-phosphate dehydrogenase (dGAPDH), and with monoclonal antibodies specific for dGAPDH (but not for the native protein). The pH-dependent properties of the conjugates have been studied using turbidimetry and light scattering. The antibody-PMAA and dGAPDH-PMAA conjugates were shown to interact with free dGAPDH and antibodies respectively as well as with each other. Insoluble aggregates of dGAPDH with antibody-PMAA and of antibodies with dGAPDH-PMAA are formed in acidic media. The same situation occurs in the mixture of antibody-PMAA and dGAPDH-PMAA: precipitation takes place in acidic media, whereas soluble associates are formed in neutral solutions. The size of the soluble associates and the number of conjugates in the associate could be regulated by pH. The competition of free dGAPDH and dGAPDH-PMAA for binding with antibody-PMAA and the dynamic release of refolded GAPDH, with no affinity to antibody-PMAA, into solution could be used for simulating chaperone action. PMID- 10832029 TI - Vanadate and copper induce overlapping oxidative stress responses in the vanadate tolerant yeast Hansenula polymorpha. AB - The mechanisms by which vanadate exerts a toxic effect on living organisms are not completely understood. This is principally due to the variety of intracellular targets of the metal and to the changes in the chemical form and oxidation states that vanadate can undergo, both in the external environment and intracellularly. In order to further elucidate the reasons for vanadate toxicity, and assuming that common detoxification mechanisms can be evoked by a general heavy metal response, we have compared some aspects of the cellular responses to vanadate and copper in the yeast Hansenula polymorpha. By means of 2D electrophoresis we show the existence of common determinants in the responses to vanadate- and copper-induced stresses. Moreover, we demonstrate that both metals induce significant increases in antioxidant enzyme levels, and that there are significant overlaps in the heavy metal and oxidative stress responses. Interestingly, vanadate induces an increase in catalase activity that is much higher than that seen with copper and, unlike copper, does not cause lipid peroxidation of cellular membranes. This suggests that H. polymorpha cells activate a further specific detoxification pathway against vanadate-induced oxidative insults. PMID- 10832030 TI - Interaction of DNA with [Cr(Schiff base)(H(2)O)(2)]ClO(4). AB - The binding of Schiff base complexes of chromium(III) of the type [Cr(salen)(H(2)O)(2)](+) and [Cr(salprn)(H(2)O)(2)](+), where salen denotes 1,2 bis(salicylideneamino)ethane and salprn denotes 1,3-bis(salicylideneamino)propane to calf thymus DNA has been investigated by absorption, emission, circular dichroism, melting temperature and viscosity measurements. These chromium(III) complexes showed absorption hyperchromicity accompanied by red shift in charge transfer band, fluorescence enhancement, increase in melting temperature, some structural changes in CD spectra and changes in specific viscosity when bound to calf thymus DNA. The binding constant K(b) has been determined from absorption measurements for both the complexes and found to be (2.5+/-0. 4)x10(3) M(-1) for [Cr(salen)(H(2)O)(2)](+) and (1.7+/-0.3)x10(4) M(-1) for [Cr(salprn)(H(2)O)(2)](+). From the binding stoichiometry of DNA [Cr(salprn)(H(2)O)(2)](+), the number of binding site size has been determined and found to be ten base pairs per bound complex molecule. The chromium(III) complexes also bring about single strand cleavage in plasmid DNA. The experimental results show that the chromium(III) complexes bind to DNA by non intercalative mode. Major groove binding is the preferred mode of interaction for these Schiff base complexes of chromium(III). PMID- 10832031 TI - Interaction of Ni(II) and Cu(II) with a metal binding sequence of histone H4: AKRHRK, a model of the H4 tail. AB - Chromatin proteins are believed to represent reactive sites for nickel binding. The unique structure of the N-terminal tail of histone H4 contains sites for post translational modification close to a histidine residue capable of anchoring binding sites for metal ions. We have analyzed as a minimal model for the H4 tail, the blocked peptide CH(3)CO-AKRHRK-CONH(2) for nickel and copper binding. Ultraviolet-visible, circular dichroism, electron paramagnetic resonance and nuclear magnetic resonance spectroscopic analysis showed that histidine acts as an anchoring metal binding site. A 1N complex is formed between pH=5-7 and 4-6 for Ni(II) and Cu(II), respectively, while at a higher pH a series of 4N complexes are formed. Above pH 8, the 2N high-spin octahedral resulted in a 4N low-spin planar Ni(II) complex. The stability constants of the Cu(II) (3N, 4N) and Ni(II) (4N) complexes with the peptide model of the H4 were distinctly higher than those for a similar blocked peptide with a histidine in the fourth position. Significant shifts in the alphaproton region in the 1H NMR spectrum of the 4N Ni complex showed that the conformation of the peptide had been dramatically affected following Ni(II) complexation. PMID- 10832032 TI - Photosensitizing activity of hematoporphyrin on Staphylococcus aureus cells. AB - The photosensitizing action of hematoporphyrin (Hp) on two Staphylococcus aureus strains was investigated to determine if the photoprocess induces in vivo damage in DNA in addition to that occurring at the level of the cytoplasmic membrane. The results obtained demonstrate that the photokilling is dependent on the Hp dose even though the two strains, having a similar Hp-binding capacity, show different levels of photosensitivity. The electrophoretic analysis of cytoplasmic membrane proteins and DNA (chromosomal and plasmidial) suggests that the membrane represents the primary target of the photoprocess, while the DNA, that is damaged both in vivo and in vitro only at relatively long irradiation time, might be a secondary target. Moreover, the photoprocess results in mutagenesis for Salmonella typhimurium tester strains. This information is particularly important in view of the potential use of photodynamic therapy for the treatment of microbial infections. PMID- 10832033 TI - Inhibition of Fas-mediated apoptosis by Trypanosoma cruzi infection. AB - Trypanosoma cruzi-infected and normal control mammalian cells were subjected to analysis of Fas-mediated apoptosis stimulated by an agonistic anti-Fas monoclonal antibody. The infected cells showed markedly hampered apoptotic changes in nuclear morphology, phosphatidylethanolamine translocation from the inside to the outside of the plasma membrane, and DNA fragmentation into multiples of 180 bp, relative to normal control cells. Upstream of these morphological and biochemical consequences, the caspase-3 activity was elevated by the Fas stimulation in a significantly greater proportion of intact control cells, but at a highly reduced rate of infected cells. The rapid elevation of caspase-8 activity in control, apoptotic cells was completely inhibited in infected cells. In an examination of the specificity of other stimulants, X-ray radiation or chemicals such as hydrogen peroxide, colchicine or etoposide did not cause significant differences in apoptotic rates between control and infected cells; tumor necrosis factor alpha, however, induced a high rate of apoptosis in control cells, with an extremely lowered rate in infected cells. This study demonstrates, for the first time, that T. cruzi infection inhibits one of the earliest steps of death receptor-mediated apoptosis, an effect that most probably involves the inhibition of caspase-8. Differential apoptotic responses in cells infected with T. cruzi and other intracellular parasites are discussed. PMID- 10832034 TI - Evidence for the regulation of beta-N-acetylhexosaminidase expression during pregnancy in the rat. AB - It is believed that the lysosomal glycohydrolase beta-N-acetylhexosaminidase plays a part in several important processes of reproduction and it has been postulated that this enzyme is subject to hormonal regulation. During pregnancy, activity levels of the enzyme are strongly increased in both human and rat serum. However, little is known about the expression of this enzyme in the female reproductive apparatus and there is no evidence linking the production of hexosaminidase alpha- and beta-subunits to pregnancy. To clarify these aspects better, we examined the enzyme activity, isoenzyme subunit composition and distribution, as well as steady state levels of alpha- and beta-subunit mRNAs in the female reproductive organs and in other selected tissues of pregnant and non pregnant rats. Among the female rat tissues tested, the ovary and kidney had the highest specific activity. Pregnancy modulated the hexosaminidase activity differently in the tissues examined. In pregnant rats, the activity decreased in the ovary but increased significantly in the uterus, liver and to a lesser extent in other tissues. The decreased hexosaminidase activity in the ovary from pregnant rats appeared to be accompanied by a disproportionately large decrease in beta-subunit mRNA abundance, whereas in the uterus and liver, an increased abundance of this transcript was detectable. The abundance of alpha-subunit mRNA was comparable in pregnant and control rat tissues. Hexosaminidase histochemical staining of tissue sections clearly demonstrates that the greatly increased activity of hexosaminidase in the uterus during pregnancy is largely due to the enzyme in the endometrium, and not to the uterus as a whole. The overall results provide evidence that, during pregnancy, a mechanism(s) of regulation of beta-N acetylhexosaminidase expression is in operation, and that the enzyme is differentially regulated in rat tissues. PMID- 10832048 TI - Research on ageing in Germany. AB - The present review on ageing research in Germany is biased towards experimental biogerontology, because this is the field which will define the future of ageing research as a whole. In absolute numbers of publications between 1995 and 1999, Germany is comparable to other large European countries. However, Germany ranks definitively last among 10 major developed countries if the numbers of scientific papers per year are seen in relation to the economic capability. This is true for the whole of biomedical research, but it is even more exaggerated for ageing research. There are potent groups in German ageing research capable of producing a good fraction of high-impact papers, however. There are many more researchers in areas highly relevant to gerontology which recently became attracted by gerontological problems. However, the importance of modern biogerontology has not made clear to decision-makers in Germany, so that structural and financial limitations will probably prevent any significant rise in the near future, which would be necessary to keep Germany along with other developed countries. PMID- 10832049 TI - Biogerontological research in Canada. AB - There is an ever increasing interest in the study of the aging process. This review is aimed to make an overview of the biological aging research in Canada. I will summarize, to the best of my knowledge, the biological aging research undertaken actually in Canadian institutions dealing with various aspects of this research using many different experimental approaches, models from animals to humans and a huge array of techniques. The biological aging research is developing continuously in Canada, however, it is very important that we assist in a near future to its huge explosion if we would respond to the needs of an ever increasing aging population. Initiatives recently proposed by the Canadian government concerning the creation of Canadian Institutes on Health Research will provide good opportunities to establish a performant, cost-effective, and innovative biological aging research. PMID- 10832050 TI - In vitro aging research in Japan. PMID- 10832051 TI - Caloric restriction and aging: an update. AB - Restricting food intake to 50 to 70% of that eaten by ad lib-fed rats and mice markedly increases longevity, retards age-associated physiological deterioration, and delays and, in some cases, prevents age-associated diseases. These actions are due to the reduced intake of calories, and thus the phenomenon has been called the antiaging action of caloric restriction (CR). This article focuses on the possible biological mechanisms underlying the antiaging action. The following three proposed mechanisms are considered in depth: 1) attenuation of oxidative damage; 2) modulation of glycemia and insulinemia; 3) hormesis. The evolution of the antiaging action of CR is also considered. Based on this consideration, a scenario unifying the above mechanisms is presented. PMID- 10832052 TI - Photoaging of the skin from phenotype to mechanisms. AB - The skin is increasingly exposed to ambient UV-irradiation thus increasing its risk for photooxidative damage with longterm detrimental effects like photoaging, which is characterized by wrinkles, loss of skin tone, and resilience. Photoaged skin displays prominent alterations in the cellular component and the extracellular matrix of the connective tissue with an accumulation of disorganized elastin and its microfibrillar component fibrillin in the deep dermis and a severe loss of interstitial collagens, the major structural proteins of the dermal connective tissue. The unifying pathogenic agents for these changes are UV-generated reactive oxygen species (ROS) that deplete and damage non enzymatic and enzymatic antioxidant defense systems of the skin. As well as causing permanent genetic changes, ROS activate cytoplasmic signal transduction pathways in resident fibroblasts that are related to growth, differentiation, senescence, and connective tissue degradation. This review focuses on the role of UV-induced ROS in the photodamage of the skin resulting in biochemical and clinical characteristics of photoaging. In addition, the relationship of photoaging to intrinsic aging of the skin will be discussed. A decrease in the overall ROS load by efficient sunscreens or other protective agents may represent promising strategies to prevent or at least minimize ROS induced photoaging. PMID- 10832053 TI - Tumor suppressors and oncogenes in cellular senescence. AB - Cyclin-dependent kinase inhibitors p16(INK4a), p21(Cip1), and p27(Kip1) are regarded as key effectors of cellular senescence. In this review, we describe three senescence-inducing pathways involving these inhibitors, namely, the p16(INK4a)/Rb pathway, the p19(ARF)/p53/p21(Cip1) pathway, and the PTEN/p27(Kip1) pathway. We emphasize the participation of tumor suppressors and oncogenes in the regulation of these senescence-inducing pathways. Finally, we discuss the impact of the Ras and Myc oncogenes on the above-mentioned pathways. PMID- 10832054 TI - Chronic caloric restriction alters muscle membrane fatty acid content. AB - Chronic caloric restriction (CR) has been demonstrated to increase longevity in lower species and studies are ongoing to evaluate its effect in higher species. A consistent metabolic feature of CR is improved insulin sensitivity and lowered lifetime glycemia, yet the mechanism responsible is currently unknown. However, the membrane's physiochemical properties, as determined by phospholipid composition, have been related to insulin action in animal and human studies and CR has been reported to alter membrane lipid content. We evaluated muscle membrane fatty acid content in rodents randomized to CR versus control diets for up to 29 months. CR was observed to increase the membrane content of C22:6 (docosahexaenoate) and to decrease C18:2 content. The membrane lipid content was related to insulin levels but not to parameters assessing glycemic control. This study suggests that membrane lipids, in particular 22:6, may contribute to the variation in insulin sensitivity seen with age. PMID- 10832055 TI - Long-term supplementation with melatonin delays reproductive senescence in rats, without an effect on number of primordial follicles. AB - The primary objective of the present experiment was to determine if lifelong supplementation with melatonin delayed reproductive senescence through decreased loss of ovarian primordial follicles. Holtzman rats were divided into three treatments on Day 10 after pupping (Day 0 = day of pupping). Treatment 1 pups had access to water, whereas Treatment 2 and 3 pups had access to water containing 10 microg/ml melatonin only at night (Treatment 2) or continuously (Treatment 3). Estrous cycles and weights of pups were monitored at selected times during the experiment; ovaries were removed for histology at 75 and 380 days of age. Vaginal opening in Treatment 2 was delayed (P <.01) compared with Treatments 1 and 3, but there was no difference (P > 0.05) among treatments in percentage of normal length estrous cycles from vaginal opening to 75 days of age. There were fewer (P < 0.001) abnormal-length estrous cycles from 180 to 380 days of age in Treatment 2 as compared with Treatments 1 or 3. There was no effect of treatment (P > 0.05) on number of primordial follicles. In conclusion, nighttime, but not continuous supplementation with melatonin, delayed puberty and reproductive senescence without any effect on number of primordial follicles. PMID- 10832056 TI - Identical longevity phenotypes are characterized by different patterns of gene expression and oxidative damage. AB - Some years ago we applied simultaneously an identical regime of selection for late-life reproduction to several normal-lived sister lines (Ra and Rb) so as to produce several selected long-lived sister lines (La and Lb). The long-lived La and Lb sister lines had statistically identical longevity phenotypes and paraquat resistance phenotypes; however, we noticed some statistically different responses of the two strains at the biochemical level. Extensive work with the La strain showed that transcriptional alterations in antioxidant gene expression are robustly associated with its extended longevity. We decided to critically test the assumption of phenotypic equivalence by subjecting the Lb strain to the same series of molecular assays as was the La strain. The two sister strains are characterized by significantly different mechanisms and patterns of antioxidant gene expression, antioxidant enzyme activity, and oxidative damage. We find that the Lb strain appears to depend on the transcriptional activation of different genes than does the La strain, and on a post-translational up-regulation of at least one other antioxidant defense gene. The phenotypic equivalence observed at the organism level need not hold at the molecular genetic level. This finding suggests that there is more than one molecular mechanism by which antioxidant defense genes can bring about an increased resistance to oxidative stress. The theoretical and empirical implications of these findings are discussed. PMID- 10832057 TI - Long-term cultured IL-2-dependent T cell lines demonstrate p16(INK4a) overexpression, normal pRb/p53, and upregulation of cyclins E or D2. AB - Acquisition of an immortal phenotype by circumvention of the normal senescence program can be an important step in tumor development and progression. The regulation of life-span checkpoints is complex and abrogation of these processes can occur at different levels. To better understand these mechanisms in long-term cultured lymphocytes we have characterized two human long-term cultured IL-2 dependent T cell lines regarding telomere length, telomerase activity, and the expression of selected cell cycle regulators (pRb, p53, cyclin E, cyclin D1, cyclin D2, cyclin D3, cdk4, p16(INK4a), p21(WAF1), p27(KIP1), c-myc, bcl-2, and NPAT). We compared these cell lines with a primary T lymphoblast population with a limited life span from the same donor. Both T cell lines with extraordinary growth capacity showed telomere length stabilization, high telomerase activity and demonstrated wild-type pattern of pRb and p53 but strong p16(INK4a) protein expression. The growth inhibitory activity of p16(INK4a) seemed to be abrogated by enhanced expression of cyclin D2, cdk4, and c-myc in one T cell line and overexpression of cyclin E in the second T cell line. PMID- 10832058 TI - Fas/APO-1 protein is increased in spaceflown lymphocytes (Jurkat). AB - Human lymphocytes flown on the Space Shuttle respond poorly to mitogen stimulation and populations of the lymphoblastoid T cell line, Jurkat, manifest growth arrest, increase in apoptosis and time- and microgravity-dependent increases in the soluble form of the cell death factor, Fas/APO-1 (sFas). The potential role of apoptosis in population dynamics of space-flown lymphocytes has not been investigated previously. We flew Jurkat cells on Space Transportation System (STS)-80 and STS-95 to determine whether apoptosis and the apparent microgravity-related release of sFas are characteristic of lymphocytes in microgravity. The effects of spaceflight and ground-based tests simulating spaceflight experimental conditions, including high cell density and low serum concentration, were assessed. Immunofluorescence microscopy showed increased cell associated Fas in flown cells. Results of STS-80 and STS-95 confirmed increase in apoptosis during spaceflight and the release of sFas as a repeatable, time dependent and microgravity-related response. Ground-based tests showed that holding cells at 1.5 million/ml in medium containing 2% serum before launch did not increase sFas. Reports of increased Fas in cells of the elderly and the increases in spaceflown cells suggest possible similarities between aging and spaceflight effects on lymphocytes. PMID- 10832059 TI - Transferrin treatment corrects aging-related immunologic and hormonal decay in old mice. AB - Experiments were conducted to study the effect of heterologous plasma transferrins separated and purified from human plasma pools on endocrine and immune functions of old, aging mice. Two similar experiments have shown that parenteral treatment with iron and zinc-free human transferrins produces a significant improvement of immunological and endocrine functions in the aging mice toward more juvenile values. Those changes occur in the thymus and its cell subsets, in peripheral blood lymphocytes, in the restoration of juvenile levels of thyroxine, in the increase of testis weight, and in the normalization of plasma zinc levels. These totally unsuspected effects of transferrin in aging mice suggest a most important role of endogenous transferrins in the maintenance of neuroendocrine and immune functions. The mechanism remains unexplained although the basic immunoenhancing and anti-apoptotic effect of transferrin vehiculated zinc may be relevant. PMID- 10832060 TI - In vitro erythrocidal activity of activated spleen cells from young and old mice. AB - We have recently reported that activated mouse spleen mononuclear cells (MNCs) efficiently lyse autologous erythrocytes in vitro (Saxena and Chandrasekhar, 2000). In the present study, we have investigated erythrocyte-depleting ability (EDA) of spleen MNCs from young and old mice. Time kinetics of survival of erythrocytes in mitogen-activated spleen cell cultures indicated that the erythrocyte depletion was significantly faster in young spleen cell cultures than in the old. Poorer EDA of old MNCs was in spite of the fact that the susceptibility to lysis actually increased in erythrocytes from old mice. Erythrocytes opsonized by a hamster anti mouse Fas monoclonal antibody, were destroyed with a much greater efficiency by young MNCs, whereas the corresponding effect of opsonization was only moderate for old MNCs. Depletion of macrophages from MNC preparations by plastic adherence as well as carbonyl-iron and magnet treatment had a marginal if any effect on EDA of young and old mouse MNCs, indicating that a lower lymphocyte-associated erythrocidal activity as one of the factors responsible for overall lower EDA associated with spleen derived MNCs of old mice. PMID- 10832061 TI - Chemical versus dual energy x-ray absorptiometry for detecting age-associated body compositional changes in male rats. AB - Aging is associated with increases in body mass and fat mass (FM), whereas fat free mass (FFM) either decreases or remains unchanged. The purpose of this study was to determine whether dual-energy X-ray absorptiometry (DXA) accurately detects age-associated changes in male Fischer 344 x Brown-Norway rats ages 8, 18, and 28 months. Eviscerated animal carcasses were first examined via the Lunar DPX-IQ DXA (small animal software version 1.0; HiRes (0.6 x 1.2 mm) medium mode). Eviscerated carcasses were then weighed, autoclaved, homogenized, and fat isolated from aliquots of homogenate via methanol/chloroform extraction. In both chemical (CHEM) and DXA analysis, carcass mass (CM), FM, and % fat were significantly higher (P < 0.0001) in the 18 and 28 versus 8-month-old rats. CHEM showed greater FFM in the 18 versus 8 months-old rats but not the 28 months-old animals. DXA was unable to detect the age-associated changes in FFM. Regression analysis showed a strong correlation between CHEM and DXA methods for CM (r = 0.98, P < 0.0001) and FM (r = 0.97, P < 0.0001), but less strong for FFM (r = 0.59, P = 0.0002). In conclusion, compared to CHEM, DXA consistently overestimated CM and FM across the age groups by 9% and 77%, respectively, and underestimated FFM by 5%. PMID- 10832062 TI - Effects of menadione and hydrogen peroxide on glutathione status in growing Escherichia coli. AB - Menadione (MD) and H2O2 caused distinct effects on glutathione status in growing Escherichia coli. Treatment of E. coli AB1157 with 1-25 mM H2O2 did not result in an appreciable decrease in intracellular total glutathione (reduced glutathione [GSH] + oxidized glutathione [GSSG]). Only when cells were treated with 25 mM H2O2 an increase in GSSG and a decrease in the GSH:GSSG ratio were observed. In cells deficient in catalase HPI, such effect was observed even at 10 mM H2O2. The exposure of E. coli AB1157 to MD caused a dose-dependent decrease in intracellular total glutathione, an increase in GSSG, and a decrease in the ratio of GSH:GSSG. In E. coli deficient in cytosolic superoxide dismutase activity, a decrease in total glutathione after incubation with 0.2 mM MD was not accompanied by an increase in GSSGin, and the ratio of GSHin:GSSGin was three times higher than in the wild-type cells. The changes in the redox status of extracellular glutathione under the action of both oxidants were similar. Although the catalase activity increased several times after exposure to both oxidants, there were little or no changes in the activity of enzymes related to glutathione metabolism. A possible role of changes in redox status of glutathione under oxidative stress is discussed. PMID- 10832063 TI - Manganese-porphyrin reactions with lipids and lipoproteins. AB - Manganese porphyrin complexes serve to catalytically scavenge superoxide, hydrogen peroxide, and peroxynitrite. Herein, reactions of manganese 5,10,15,20 tetrakis(N-ethylpyridinium-2-yl)porphyrin (MnTE-2-PyP(5+)) with lipids and lipid hydroperoxides (LOOH) are examined. In linoleic acid and human low-density lipoprotein (LDL), MnTE-2-PyP(5+) promotes oxidative reactions when biological reductants are not present. By redox cycling between Mn(+3) and Mn(+4) forms, MnTE-2-PyP(5+) initiates lipid peroxidation via decomposition of 13(S)hydroperoxyoctadecadienoic acid [13(S)HPODE], with a second-order rate constant of 8.9 x 10(3) M(-1)s(-1)and k(cat) = 0.32 s(-1). Studies of LDL oxidation demonstrate that: (i) MnTE-2-PyP(5+) can directly oxidize LDL, (ii) MnTE-2-PyP(5+) does not inhibit Cu-induced LDL oxidation, and (iii) MnTE-2 PyP(5+) plus a reductant partially inhibit lipid peroxidation. MnTE-2-PyP(5+) (1 5 microM) also significantly inhibits FeCl(3) plus ascorbate-induced lipid peroxidation of rat brain homogenate. In summary, MnTE-2-PyP(5+) initiates membrane lipid and lipoprotein oxidation in the absence of biological reductants, while MnTE-2-PyP(5+) inhibits lipid oxidation reactions initiated by other oxidants when reductants are present. It is proposed that, as the Mn(+3) resting redox state of MnTE-2-PyP(5+) becomes oxidized to the Mn(+4) redox state, LOOH is decomposed to byproducts that propagate lipid oxidation reactions. When the manganese of MnTE-2-PyP(5+) is reduced to the +2 state by biological reductants, antioxidant reactions of the metalloporphyrin are favored. PMID- 10832064 TI - EPR spin trapping detection of carbon-centered carotenoid and beta-ionone radicals. AB - Free radical intermediates were detected by the electron paramagnetic resonance spin trapping technique upon protonation/deprotonation reactions of carotenoid and beta-ionone radical ions. The hyperfine coupling constants of their spin adducts obtained by spectral simulation indicate that carbon-centered radicals were trapped. The formation of these species was shown to be a result of chemical oxidation of neutral compounds by Fe(3+) or I(2) followed by deprotonation of the corresponding radical cations or addition of nucleophilic agents to them. Bulk electrolysis reduction of beta-ionone and carotenoids also leads to the formation of free radicals via protonation of the radical anions. Two different spin adducts were detected in the reaction of carotenoid polyenes with piperidine in the presence of 2-methyl-2-nitroso-propane (MNP). One is attributable to piperidine radicals (C(5)H(10)N*) trapped by MNP and the other was identified as trapped neutral carotenoid (beta-ionone) radical produced via protonation of the radical anion. Formation of these radical anions was confirmed by ultraviolet visible spectroscopy. It was found that the ability of carotenoid radical anions/cations to produce neutral radicals via protonation/deprotonation is more pronounced for unsymmetrical carotenoids with terminal electron-withdrawing groups. This effect was confirmed by the radical cation deprotonation energy (H(D)) estimated by semiempirical calculations. The results indicate that the ability of carotenoid radical cations to deprotonate decreases in the sequence: beta-ionone > unsymmetrical carotenoids > symmetrical carotenoids. The minimum H(D) values were obtained for proton abstraction from the C(4) atom and the C(5) methyl group of the cyclohexene ring. It was assumed that deprotonation reaction occurs preferentially at these positions. PMID- 10832065 TI - Nitrite-induced deamination and hypochlorite-induced oxidation of DNA in intact human respiratory tract epithelial cells. AB - No modification of purine or pyrimidine bases was observed when isolated DNA was incubated with 1 mM nitrite at pH 7.4. However, exposure of human bronchial epithelial cells in culture medium at pH 7.4 to nitrite at concentrations of 100 microM or greater led to deamination of purine bases in cellular DNA. Deamination was more extensive in cells exposed to lower extracellular pH values and higher nitrite concentrations. Significant increases in the levels of xanthine and hypoxanthine, putative deamination products of guanine and adenine, respectively, were observed in DNA from nitrite-treated cells but no rise in any base oxidation products such as 8-hydroxyguanine. This pattern of damage suggests that exposure of cells to nitrite (even at pH 7.4) leads to intracellular generation of "reactive nitrogen species" capable of deaminating purines in DNA. In addition, significant DNA strand breakage occurred in nitrite-treated cells. The time course of base damage suggested that the repair of deaminated purine lesions in these cells is slow. By contrast, DNA isolated from cells exposed to hypochlorous acid (HOCl) has significant oxidation of pyrimidine bases and chlorination of cytosine but little oxidation of purines. Exposure of cells to both species (NO(2)(-) plus HOCl) potentiated the oxidative DNA base damage observed but decreased the extent of deamination. We hypothesize that this is due to the formation of nitryl chloride (NO(2)Cl) from reaction of HOCl with *NO(2)(-). The relevance of our observations to events in the stomach and respiratory tract, at sites of inflammation, and in ischemic tissues is discussed. PMID- 10832066 TI - Kinetics of phycocyanine bilin groups destruction by peroxyl radicals. AB - Bilin groups in c-phycocyanine are readily bleached by peroxyl radicals produced in the thermolysis of 2, 2'-azobis(2-amidinopropane). From an evaluation of the bilin groups destroyed per radical that interacts with the protein, it is concluded that the bilin moiety is the main target of the radicals. Kinetic expressions are derived that allows an estimation of the substrate reactivity from the analysis of the rate of bilin group modification as a function of the protein concentration. From this analysis it is concluded that micromolar concentrations of c-phycocyanine are able to reduce the steady state concentration of the peroxyl radicals by one half, indicating a high antioxidant activity for this compound. This conclusion is confirmed by measuring the capacity of the protein to protect 1-naphthol from modification by peroxyl radicals. The results obtained show that the bilin groups have, on a molar basis, an antioxidant activity similar to that of potent antioxidants such as catechin. PMID- 10832067 TI - Bioavailability and metabolism of the flavonol quercetin in the pig. AB - During the last years, much data pointing to putative health-promoting effects of dietary plant-derived flavonoids (stemming mainly from epidemiological and in vitro studies) have been published. Our knowledge, however, concerning the systemic availability of these substances after ingestion with food is only sketchy. In the present study, we have investigated the bioavailability of the flavonol quercetin after intravenous and oral application in pigs equipped with a permanent jugular catheter. Each animal received a single intravenous dose of quercetin (0.4 mg/kg body weight) and one week later an oral dose of 50 mg/kg. A single animal additionally received an oral dose of 500 mg/kg one week after the lower oral dose. Blood samples were drawn at defined intervals over a total period of three days following the application of quercetin. Analysis of quercetin and some of its metabolites (isorhamnetin, tamarixetin, kaempferol) in plasma samples were performed by HPLC. The calculated apparent bioavailability of free, unchanged quercetin after intake of 50 mg quercetin/kg body weight was 0.54+/-0.19%. Bioavailability was, however, considerably increased to 8.6+/-3.8% after additionally taking into account conjugated quercetin and further increased to 17.0+/-7.1% by including quercetin's metabolites. Our results further indicate, that the conjugation of orally administered quercetin with glucuronic and sulfuric acid appears to preferentially occur in the intestinal wall. PMID- 10832068 TI - Tetradecylthioacetic acid and tetradecylselenoacetic acid inhibit lipid peroxidation and interact with superoxide radical. AB - Reactive oxygen species are thought to induce cellular damage and to play a pathological role in several human diseases. Tetradecylthioacetic acid (TTA) was previously reported to prevent the oxidative modification of low-density lipoprotein (LDL) particles and to act as an antioxidant. In this study we present a new fatty acid analogue, namely tetradecylselenoacetic acid (TSA), in which the sulfur atom of TTA is replaced by a selenium atom. TSA was more potent than TTA in increasing the lag time before the onset of LDL oxidation and this effect was dose dependent. TTA and TSA were shown to reduce the iron-ascorbate induced microsomal lipid peroxidation, TSA being more efficient than TTA. TTA and TSA, in the presence of iron, interacted with the superoxide radical as assessed by direct and indirect testing methods. TSA like TTA failed to scavenge 1.1 diphenyl-2-picrylhydrazyl radicals. TSA bound copper ions as shown by the wavelength spectra measurement. These results suggest that TTA and TSA exert their antioxidant capacity by interaction with copper or iron ions in radical scavenging, TSA being more potent than TTA. Nevertheless, a chelating effect resulting in chemically inactive metal ions cannot be excluded. PMID- 10832069 TI - Do spin traps also act as classical chain-breaking antioxidants? A quantitative kinetic study of phenyl tert-butylnitrone (PBN) in solution and in liposomes. AB - Free radical spin traps such as phenyl tert-butylnitrone (PBN) are often reported to provide protection of the central nervous system of animal models against free radical damage, and the effects are attributed to its "antioxidant activity." The effects of PBN and p-CH(3)O-PBN were compared with known antioxidants, alpha tocopherol and 2,2,5,7,8-pentamethyl-6-hydroxychroman (PMHC), in quantitative kinetic studies of lipid peroxidation thermally initiated under controlled conditions. Results obtained on the spin traps in organic solvents and in dilinoleoyl phosphatidylcholine (DLPC) bilayers indicated that the spin traps do not act as peroxyl radical trapping antioxidants but rather act only as moderate "retarders" of oxygen uptake at relatively high concentration. At low oxygen partial pressures, e.g., 14 torr, which better reflect oxygen partial pressures in biological systems, PBN provides a more significant reduction in oxygen uptake (up to 50%) by DLPC bilayers but still did not act as a typical antioxidant. However, at low partial pressures, PBN does act cooperatively with PMHC. It is suggested that its role in biological fluids and tissues may be to extend the suppressed oxidation by natural antioxidants expected to be present. The combination of antioxidant/spin trap, alpha-(3, 5-di-tert-butyl-4-hydroxyphenyl) N-tert-butylnitrone did not exhibit any enhanced antioxidant efficiency compared with the related hindered phenol, 2,6-di-tert-butyl-4-methoxyphenol. PMID- 10832070 TI - Zinc deficiency induces oxidative stress and AP-1 activation in 3T3 cells. AB - It has been postulated that one mechanism underlying zinc deficiency-induced tissue alterations is excessive cellular oxidative damage. In the present study we investigated if zinc deficiency can induce oxidative stress in 3T3 cells and trigger select intracellular responses that have been associated to oxidative stress. Cells were exposed to control media or to chelated media containing 0.5, 5, or 50 microM zinc for 24 or 48 h. The oxidative status of the cells was evaluated as an increase in the fluorescence of the probe 5(or 6)-carboxy-2'7' dichlorodihydrofluorescein diacetate (DCDCDHF). After 24 and 48 h of exposure, the fluorescence intensity was significantly higher (4- to 15-fold) in the 0.5 and 5 microM Zn groups compared to the 50 microM Zn and control groups. The activity of the antioxidant enzymes CuZn (CuZnSOD) and Mn (MnSOD) superoxide dismutases was significantly higher in the 0.5 and 5 microM Zn cells compared to the 50 microM Zn and control groups at both the 24 and 48 h time points. These higher activities were associated with higher levels of MnSOD mRNA. After 24 h in culture, the level of activated AP-1 was markedly higher in the 0.5 and 5 microM Zn cells than in the control (72 and 58%, respectively) and 50 microM Zn cells (73 and 60%, respectively). NF-kappaB binding activity was lower in the 0.5 and 5 microM Zn cells than in controls. Thus, oxidative stress is induced by zinc deficiency in 3T3 cells. This oxidative stress results in an upregulation of oxidant defense mechanisms. PMID- 10832071 TI - Coenzyme Q improves LDL resistance to ex vivo oxidation but does not enhance endothelial function in hypercholesterolemic young adults. AB - Oxidative modification of low-density lipoprotein (LDL) may cause arterial endothelial dysfunction in hyperlipidemic subjects. Antioxidants can protect LDL from oxidation and therefore improve endothelial function. Dietary supplementation with coenzyme Q (CoQ(10)) raises its level within LDL, which may subsequently become more resistant to oxidation. Therefore, the aim of this study was to assess whether oral supplementation of CoQ(10) (50 mg three times daily) is effective in reducing ex vivo LDL oxidizability and in improving vascular endothelial function. Twelve nonsmoking healthy adults with hypercholesterolemia (age 34+/-10 years, nine women and three men, total cholesterol 7.4+/-1.1 mmol/l) and endothelial dysfunction (below population mean) at baseline were randomized to receive CoQ(10) or matching placebo in a double-blind crossover study (active/placebo phase 4 weeks, washout 4 weeks). Flow-mediated (FMD, endothelium dependent) and nitrate-mediated (NMD, smooth muscle-dependent) arterial dilatation were measured by high-resolution ultrasound. CoQ(10) treatment increased plasma CoQ(10) levels from 1.1 +/-0.5 to 5.0+/-2.8 micromol/l (p =.009) but had no significant effect on FMD (4.3+/-2.4 to 5.1+/-3.6 %, p =.99), NMD (21.6+/-6.1 to 20.7+/-7.8 %, p = .38) or serum LDL-cholesterol levels (p = .51). Four subjects were selected randomly for detailed analysis of LDL oxidizability using aqueous peroxyl radicals as the oxidant. In this subgroup, CoQ(10) supplementation significantly increased the time for CoQ(10)H(2) depletion upon oxidant exposure of LDL by 41+/-19 min (p = .04) and decreased the extent of lipid hydroperoxide accumulation after 2 hours by 50+/-37 micromol/l (p =.04). We conclude that dietary supplementation with CoQ(10) decreases ex-vivo LDL oxidizability but has no significant effect on arterial endothelial function in patients with moderate hypercholesterolemia. PMID- 10832072 TI - Acute methanol intoxication generates free radicals in rats: an ESR spin trapping investigation. AB - Electron spin resonance (ESR) spectroscopy has been used to investigate free radical generation in rats with acute methanol poisoning. The spin trapping technique was used where a spin trapping agent, alpha-(4-pyridyl 1-oxide)-N-tert butylnitrone (POBN), reacted with the corresponding alcohol-derived or alcohol dependent radical to form radical adducts. One radical adduct was detected in both bile and urine samples 2 h after acute methanol poisoning in male Sprague Dawley rats. The hyperfine coupling constants for the radical adduct from [(13)C] labeled methanol detected in the bile were a(N) = 15.58, a(beta)(H) = 2.81 G, and a(beta)(13C) = 4.53 G, which unambiguously identified this species as POBN/*CH@OH. The same radical adduct was detected in urine. The identification of a methanol-derived radical adduct in samples from bile and urine provided strong direct evidence for the generation of the alcohol-derived radicals during acute intoxication by methanol. Simultaneous administration of the alcohol dehydrogenase inhibitor 4-methylpyrazole and methanol resulted in an increase in the generation of the free radical metabolite detected in the bile. This is the first ESR evidence of methanol-derived free radical generation in an animal model of acute methanol intoxication. PMID- 10832073 TI - Dual role of glutathione in selenite-induced oxidative stress and apoptosis in human hepatoma cells. AB - It is well known that glutathione, the major intracellular antioxidant, is closely involved in the metabolism and bioactivity of selenium. In the present study, glutathione was demonstrated to play a dual role on selenite (Se)-induced oxidative stress and apoptosis in human hepatoma HepG(2) cells. The experiment was carried out in two different modes to modulate intracellular reduced glutathione (GSH) content. In Mode A (pretreatment), cells were pretreated with N acetylcysteine (NAC), buthionine sulfoximine (BSO), or GSH prior to Se exposure. In Mode B (simultaneous treatment), cells were treated with Se and NAC, BSO, or GSH simultaneously. It was found that Se-induced oxidative stress and apoptosis are closely related to the intracellular level of GSH. Both the increase and depletion of GSH content significantly enhanced Se-induced oxidative stress and apoptosis in HepG(2) cells. Results from this study clearly demonstrated that GSH has a dual role in the effects of Se on cancer cells: (i) GSH acts as a pro oxidant, facilitating Se-induced oxidative stress, and (ii) GSH acts as an antioxidant, protecting against Se-induced oxidative stress and apoptosis. Understanding such a unique association between GSH and Se may help to explain the controversy in the literature over the complex relationship between selenium and glutathione, and ultimately the capability of selenium to prevent cancer. PMID- 10832074 TI - Exposure to cigarette smoke increases apoptosis in the rat gastric mucosa through a reactive oxygen species-mediated and p53-independent pathway. AB - Cigarette smoking is a major risk factor for gastric cancer and peptic ulcer. The aim of our study was to investigate the relationship between exposure to cigarette smoke and apoptosis in the rat gastric mucosa and the mechanism involved. Rats were exposed to different concentrations of cigarette smoke (0, 2, and 4%) once daily for a different number of 1 h periods (1, 3, 6, and 9 d). Apoptosis was identified by the terminal deoxy-transferase (TdT)-mediated dUTP biotin nick end labeling (TUNEL) method and caspase-3 activity. The mucosal xanthine oxidase (XO) activity and p53 level were also measured. The results showed that exposure to cigarette smoke produced a time- and concentration dependent increase in apoptosis in the rat gastric mucosa that was accompanied by an increase in XO activity. The increased apoptosis and XO activity could be detected after even a single exposure. In contrast, the level of p53 was elevated only in the later stage of cigarette smoke exposure. The apoptotic effect could be blocked by pretreatment with an XO inhibitor (allopurinol, 20 mg/kg intraperitoneally) or a hydroxyl free radical scavenger (DMSO, 0.2%, 1 ml/kg intravenously). However, neither of these treatments had any effect on the p53 level of the mucosa. In summary, we conclude that exposure to cigarette smoke can increase apoptosis in the rat gastric mucosa through a reactive oxygen species- (ROS) mediated and a p53-independent pathway. PMID- 10832075 TI - Oxygen-insensitive nitroreductases of Escherichia coli do not reduce 3 nitrotyrosine. AB - The oxygen-insensitive nitroreductases nfsA and nfsB are known to reduce para nitrated aromatic compounds. We tested the hypothesis that these nitroreductases are capable of reducing 3-nitrotyrosine in proteins and peptides, as well as in free amino acids using wild-type and nfsA nfsB mutant strains of Escherichia coli. E. coli homogenates were incubated with nitrated proteins and the level of 3-nitrotyrosine immunoreactivity was assayed by Western blotting. Assay conditions that allow the nitroreductases to rapidly reduce nitrofurantoin did not result in the modification of 3-nitrotyrosine in protein, peptide, or free amino acid. Stimulation of nfsA nfsB activity with paraquat had no effect on 3 nitrotyrosine reduction. Nonlethal exposure of E. coli to peroxynitrite/CO(2) resulted in the reproducible nitration of tyrosine residues in endogenous proteins. The degree of 3-nitrotyrosine immunoreactivity over the 2-h postexposure period did not differ between mutant and wild-type strains. These results indicate that the nfsA and nfsB enzymes do not reduce 3-nitrotyrosine. PMID- 10832077 TI - Role of antioxidant enzyme expression in the selective cytotoxic response of glioma cells to gamma-linolenic acid supplementation. AB - We hypothesized that the cytotoxic effect of GLA observed in glioma but not normal glial cells reflects differences in GLA metabolism and/or antioxidant enzyme levels between these cells. The PUFA content of unsupplemented glioma cells was approximately 50% of that seen in unsupplemented astrocytes. Supplementation with 20 microM GLA for 24 h led to a 230 and 22% increase in glioma and astrocyte PUFA content, respectively, such that both supplemented cell types contained similar levels of PUFA. No major differences were seen in terms of GLA metabolites retained in the cells or secreted into the media following incubation with [(3)H]-GLA. No significant differences were observed in activity of MnSOD or CuZn-SOD between the cells. However, CAT and GPx activity in the glioma cells was significantly higher and lower, respectively, than observed in normal astrocytes. GLA supplementation resulted in a significant increase in CAT activity in normal astrocytes; glioma CAT activity was unchanged. No significant change was seen in the other antioxidant enzymes following GLA supplementation. These results suggest that the cytotoxic effect of GLA on glioma cells reflects both increased PUFA content and an inability to upregulate CAT. PMID- 10832076 TI - Decreased serum concentrations of nitric oxide metabolites among Chinese in an endemic area of chronic arsenic poisoning in inner Mongolia. AB - Prolonged exposure to arsenic results in peripheral and cardiovascular manifestations, as does impaired production of endothelial nitric oxide (NO). In vitro studies have indicated that endothelial cells undergo damage by arsenic. However, no information has been available on the relationship between NO synthesis and chronic arsenic poisoning in humans. The present study was designed to reveal this question. The subjects were 33 habitants who continued to drink well water containing high concentrations of inorganic arsenic (mean value = 0.41 microg/ml) for about 18 years in Inner Mongolia, China, and 10 other people who lived in this area but exposed to minimal concentrations of arsenic (mean value = 0.02 microg/ml) were employed as controls. Mean blood concentration of total arsenic was six times higher in exposed subjects than controls; 42.1 vs. 7.3 ng/ml, p <.001. Mean serum concentration of nitrite/nitrate, stable metabolites of endogenous NO, was lower in arsenic-exposed subjects than in controls: 24.7 vs. 51.6 microM, p<.001. In total samples, an inverse correlation with serum nitrite/nitrate levels was strong for blood inorganic arsenic (r = -0.52, p <.001) and less strong for its metabolites, monomethyl arsenic (r = -0.45, p<.005) and dimethyl arsenic (r = -0.37, p<.05). Furthermore, serum nitrite/nitrate concentration was significantly correlated with nonprotein sulfhydryl level in whole blood (r = 0.58, p<.001). In an in vitro study, we demonstrated that inorganic arsenite or arsenate suppresses the activity of endothelial NO synthase in human umbilical vein endothelial cells. These results suggest that long-term exposure to arsenic by drinking well water possibly reduces NO production in endothelial cells, resulting in a decrease in reduced nitrite/nitrate concentrations. Peripheral vascular disorders caused by arsenic may be attributable in part to impairment of NO production in vivo. PMID- 10832078 TI - Synthesis and preliminary evaluation of (18)F-labeled 4-thia palmitate as a PET tracer of myocardial fatty acid oxidation. AB - Interest remains strong for the development of a noninvasive technique for assessment of regional fatty acid oxidation rate in the myocardium. (18)F-labeled 4-thia palmitate (FTP, 16-[(18)F]fluoro-4-thia-hexadecanoic acid) has been synthesized and preliminarily evaluated as a metabolically trapped probe of myocardial fatty acid oxidation for positron emission tomography (PET). The radiotracer is synthesized by Kryptofix 2.2.2/K(2)CO(3) assisted nucleophilic radiofluorination of an iodo-ester precursor, followed by alkaline hydrolysis and by purification by reverse phase high performance liquid chromatography. Biodistribution studies in rats showed high uptake and long retention of FTP in heart, liver, and kidneys consistent with relatively high fatty acid oxidation rates in these tissues. Inhibition of carnitine palmitoyl-transferase-I caused an 80% reduction in myocardial uptake, suggesting the dependence of trapping on the transport of tracer into the mitochondrion. Experiments with perfused rat hearts showed that the estimates of the fractional metabolic trapping rate (FR) of FTP tracked inhibition of oxidation rate of palmitate with hypoxia, whereas the FR of the 6-thia analog 17-[(18)F]fluoro-6-thia-heptadecanoic acid was insensitive to hypoxia. In vivo defluorination of FTP in the rat was evidenced by bone uptake of radioactivity. A PET imaging study with FTP in normal swine showed excellent myocardial images, prolonged myocardial retention, and no bone uptake of radioactivity up to 3 h, the last finding suggesting a species dependence for defluorination of the omega-labeled fatty acid. The results support further investigation of FTP as a potential PET tracer for assessing regional fatty acid oxidation rate in the human myocardium. PMID- 10832079 TI - High specific radioactivity (1R,2S)-4-[(18)F]fluorometaraminol: a PET radiotracer for mapping sympathetic nerves of the heart. AB - The radiolabeled catecholamine analogue (1R, 2S)-6-[(18)F]fluorometaraminol (6 [(18)F]FMR) is a substrate for the neuronal norepinephrine transporter. It has been used as a positron emission tomography (PET) ligand to map sympathetic nerves in dog heart. 6-[(18)F]FMR could be only synthesized with low specific radioactivity, which precluded its use in human subjects. We have recently prepared (1R,2S)-4-[(18)F]fluorometaraminol (4-[(18)F]FMR), a new fluoro-analogue of metaraminol, with high specific radioactivity (56-106 GBq/micromol). In the present study, we demonstrate in rats that 4-[(18)F]FMR possesses similar affinity toward myocardial norepinephrine transport mechanisms as 6-[(18)F]FMR. When compared with control animals, an 80% and 76% reduction in myocardial uptake was observed in animals pretreated with desipramine (an inhibitor of the neuronal norepinephrine transporter) and with reserpine (a blocker of the vesicular storage of monoamines), respectively. The entire radioactivity in rat myocardium represented unmetabolized parent tracer as determined by high performance liquid chromatography analysis of tissue extracts. In dogs, myocardial kinetics of 4 [(18)F]FMR were assessed using PET. A rapid and high uptake was observed, followed by prolonged cardiac retention. A heart-to-lung ratio of 15 was reached 10 min after injection of the radiotracer. Pretreatment with desipramine reduced the heart half-life of 4-[(18)F]FMR by 90% compared with control. Moreover, an infusion of tyramine caused a rapid decline of radioactivity in the heart. This demonstrates that 4-[(18)F]FMR specifically visualizes sympathetic neurons in dog heart. High specific radioactivity 4-[(18)F]FMR is a promising alternative to 6 [(18)F]FMR for myocardial neuronal mapping with PET in humans. PMID- 10832080 TI - Comparison of N-[(11)C]methyl-norchloroepibatidine and N-[(11)C]methyl-2-(2 pyridyl)-7-azabicyclo[2.2.1]heptane with N-[(11)C]methyl-epibatidine in small animal PET studies. AB - Structural variations of the nicotinic acetylcholine receptor radioligand N [(11)C]methyl-epibatidine were made to form (11)C-labeled N-methyl norchloroepibatidine (N-methyl-NorchloroEPB) and N-methyl-2-(2-pyridyl)-7 azabicyclo[2.2.1]heptane (N-methyl-2PABH). Radiosyntheses were performed by methylation with high radiochemical purities (>98%) and with specific activities between 140 and 500 GBq/micromol at the end of synthesis. The radiochemical yield (decay-corrected, related to [(11)C]CH(3)I) was between 5 and 10%. Positively and negatively radiolabeled enantiomers were prepared in high optical purity (>98%ee) by labeling of the appropriate optically active substrates, which were obtained via chiral high performance liquid chromatography. For in vivo studies radioligands were administered intravenously in rats. Brain uptake curves were acquired and combined with blocking experiments. Brain uptake of N-[(11)C]methyl NorchloroEPB was similar to that of N-[(11)C]methyl-EPB whereas N-[(11)C]methyl 2PABH with the modified pyridine ring had a significantly lower uptake. PMID- 10832081 TI - 2-((2-((dimethylamino)methyl)phenyl)thio)-5-iodophenylamine (ADAM): an improved serotonin transporter ligand. AB - Serotonin transporters (SERT) are target-sites for commonly used antidepressants, such as fluoxetine, paroxetine, sertraline, and so on. Imaging of these sites in the living human brain may provide an important tool to evaluate the mechanisms of action as well as to monitor the treatment of depressed patients. Synthesis and characterization of an improved SERT imaging agent, ADAM (2-((2 ((dimethylamino)methyl)phenyl)thio)-5-iodophenylamine)(7) was achieved. The new compound, ADAM(7), displayed an extremely potent binding affinity toward SERT ( K(i)=0.013 nM, in membrane preparations of LLC-PK(1)-cloned cell lines expressing the specific monoamine transporter). ADAM(7) also showed more than 1,000-fold selectivity for SERT over norepinephrine transporter (NET) and dopamine transporter (DAT) ( K(i)=699 and 840 nM, for NET and DAT, respectively). The radiolabeled compound [(125)I]ADAM(7) showed an excellent brain uptake in rats (1.41% dose at 2 min post intravenous [IV] injection), and consistently displayed the highest uptake (between 60-240 min post IV injection) in hypothalamus, a region with the highest density of SERT. The specific uptake of [(125)I]ADAM(7) in the hypothalamus exhibited the highest target-to-nontarget ratio ([hypothalamus - cerebellum]/cerebellum was 3.97 at 120 min post IV injection). The preliminary imaging study of [(123)I]ADAM in the brain of a baboon by single photon emission computed tomography (SPECT) at 180-240 min post IV injection indicated a specific uptake in midbrain region rich in SERT. These data suggest that the new ligand [(123)I]ADAM(7) may be useful for SPECT imaging of SERT binding sites in the human brain. PMID- 10832082 TI - In vivo evaluation of [(11)C]SA4503 as a PET ligand for mapping CNS sigma(1) receptors. AB - The potential of the (11)C-labeled selective sigma(1) receptor ligand 1-(3,4 dimethoxyphenethyl)-4-(3-phenylpropyl)piperazine ([(11)C]SA4503) was evaluated in vivo as a positron emission tomography (PET) ligand for mapping sigma(1) receptors in rats. SA4503 is known to have a high affinity (IC(50) = 17.4 nM) and a higher selectivity (sigma(1)/sigma(2) = 103) for the sigma(1) receptor. A high and increasing brain uptake of [(11)C]SA4503 was found. Pre-, co- and postinjection of cold SA4503 significantly decreased uptake of [(11)C]SA4503 in the brain, spleen, heart, lung, and kidney in which sigma receptors are present as well as in the skeletal muscle. In the blocking study with one of four sigma receptor ligands including haloperidol, (+)-pentazocine, SA4503, and (-) pentazocine (in the order of their affinity for sigma(1) receptor subtype), SA4503 and haloperidol significantly reduced the brain uptake of [(11)C]SA4503 to approximately 30% of the control, but the other two benzomorphans did not. A high specific uptake of [(11)C]SA4503 by the brain was also confirmed by ex vivo autoradiography (ARG) and PET. Ex vivo ARG showed a higher uptake in the vestibular nucleus, temporal cortex, cingulate cortex, inferior colliculus, thalamus, and frontal cortex, and a moderate uptake in the parietal cortex and caudate putamen. Peripherally, the blocking effects of the four ligands depended on their affinity for sigma(1) receptors. No (11)C-labeled metabolite was detected in the brain 30 min postinjection, whereas approximately 20% of the radioactivity was found as (11)C-labeled metabolites in plasma. These results have demonstrated that the (11)C-labeled sigma(1) receptor ligand [(11)C]SA4503 has a potential for mapping sigma(1) receptors in the central nervous system and peripheral organs. PMID- 10832083 TI - Syntheses and biological evaluation of (18)F-labeled 3-(1-benzyl-piperidin-4-yl) 1-(1-methyl-1H-indol-3-yl)propan-1-ones for in vivo mapping of acetylcholinesterase. AB - We synthesized novel (18)F-labeled acetylcholinesterase (AChE) inhibitors, 3-[1 (3- and 4-[(18)F]fluoromethylbenzyl)piperidin-4-yl]-1-(1-methyl-1H-i ndol-3-yl )propan-1-ones ([(18)F]1 and [(18)F]2) and 3-[1-(4-[(18)F]fluorobenzyl)piperidin 4-yl]-1-(1-methyl-1H-i ndol-3-yl )propan-1-one ([(18)F]3) in high yields (decay corrected, 25%-40%) and with high effective specific activities (>37 GBq/micromol). Tissue distribution studies of the [(18)F]1 and the [(18)F]3 in mice showed the nonspecific bindings in brain regions, with metabolic defluorination of the [(18)F]1. The result suggests that these radioligands may not be suitable agents for in vivo mapping of AChE, despite their potent in vitro anti-AChE activities. PMID- 10832084 TI - Radiochemical synthesis and tissue distribution of Tc-99m-labeled 7alpha substituted estradiol complexes. AB - The diagnosis and staging of breast cancer could be improved by the development of radiopharmaceutical imaging agents that provide a noninvasive determination of the estrogen receptor (ER) status of tumor cells. Agents labeled with (99m)Tc would be especially valuable in this regard. In attempting to achieve this goal, we synthesized four (99m)Tc-labeled 7alpha-substituted estradiol complexes. One complex utilizes the "3+1" mixed ligand design to introduce the Tc metal, whereas the other three took advantage of the cyclopentadienyltricarbonylmetal (CpTM) design. The Tc moieties were attached to the 7alpha position of estradiol with a hexyl tether, a monoether tether, or a polyether tether. The corresponding rhenium compounds have binding affinities for the ER of 20-45% compared with estradiol. Radiochemical yields of the (99m)Tc-labeled compounds ranged from approximately 15% for the CpT-Tc complexes to 95% for the 3 + 1 inorganic complex. Tissue distribution studies in immature female rats showed low nonreceptor-mediated uptake in the target organs and high uptake in nontarget organs such as the liver and fat. These complexes represent the first time that estradiol has been labeled at the 7alpha position with (99m)Tc and provide a further refinement of our understanding of ligand structure-binding affinity correlations for the ER. PMID- 10832085 TI - Synthesis and evaluation of a new series of 17alpha-[(123)I]iodovinyl estradiols. AB - A series of 17alpha-substituted estradiols was synthesized in which the stereochemical characteristics of carbons 20 and 21 were modified. It was found that the (Z)-isomer demonstrated more favorable receptor binding affinity than the corresponding (E)-isomer. PMID- 10832086 TI - Extending the lifetime of anticoagulant oligodeoxynucleotide aptamers in blood. AB - We have investigated (123)I and (125)I DNA aptamer analogs of anticoagulant DNA aptamers to thrombin exosite 1 and exosite 2 for thrombus imaging potential. Two severe problems are rapid clearance from circulating blood and blood nuclease. With aptamers (unlike antisense) the nucleotide analogs used in polymerase chain reaction-selection cycles also must be used in the radiotracer. We investigated 3'-biotin-streptavidin (SA) bioconjugates of the aptamers to alleviate these problems. Blood nuclease assays and biodistribution analysis were used in the mouse and rabbit. We found that 3'-biotin protected the aptamers significantly from blood nuclease in vitro, but it did not slow in vivo clearance. In contrast, the 3'-biotin-SA bioconjugates were resistant to blood nuclease in vitro and were also longer-lived (10-20 times) in vivo. Bioconjugate aptamers retained affinity for thrombin. Two solutions emerge: 1) In noncirculating blood (within a thrombus) 3'-biotin extends aptamer lifetime, whereas 2) in circulating blood (the transport medium), where more aggressive clearance is encountered, 3'-SA extends aptamer lifetime. PMID- 10832087 TI - Involvement of the glutathione S-conjugate compounds and the MRP protein in Tc 99m-tetrofosmin and Tc-99m-sestamibi uptake in glioma cell lines. AB - The objective of this study was to compare the accumulation of Tc-99m-tetrofosmin and Tc-99m-sestamibi in four grade IV glioma cell lines and to correlate their accumulation with the multidrug resistance of the cells. Tc-99m-tetrofosmin in all glioma cell lines showed slightly higher uptake and more efficient release beyond 150 min than Tc-99m-sestamibi and the retention of both tracers in the cells was to a certain extend inversely proportional to their degree of multidrug resistance. The results obtained showed that the efflux of both tracers was carried out only in part through the MRP/GS-X pump system. Tc-99m-tetrofosmin showed good potential as a marker of recurrent malignant glioma and in vivo studies are currently underway to confirm these observations. PMID- 10832088 TI - An alternative approach to the preparation of (188)Re radiopharmaceuticals from generator-produced [(188)ReO(4)](-): efficient synthesis of (188)Re(V)-meso-2,3 dimercaptosuccinic acid. AB - A new efficient approach for the preparation of (188)Re radiopharmaceuticals starting from [(188)ReO(4)](-), produced at a carrier-free level through the (188)W/(188)Re generator system, is described. The reaction procedure was based on the combined action of different reagents and has been applied in detail to the preparation of the therapeutic agent (188)Re(V)-DMSA (H(2)DMSA [meso-2,3 dimercaptosuccinic acid]). The most efficient combination required the use of SnCl(2), oxalate ions, and gamma-cyclodextrin. These were reacted with [(188)ReO(4)](-) and H(2)DMSA to afford the final radiopharmaceutical in high radiochemical purity, at room temperature, and in weakly acidic solution. The role played by the various reagents in the reaction was investigated. It was found that SnCl(2) behaved as the actual reducing agent, whereas oxalate and gamma-cyclodextrin greatly enhanced the ease of reduction of [(188)ReO(4)](-) through the action of two hypothetical mechanisms. In the first step of the reaction, oxalate ions gave rise to the formation of Re(VII) complexes with the concomitant expansion of the coordination sphere of the metal. This process strongly favored the electron transfer between Sn(2+) and Re(+7) centers, giving rise to intermediate reduced rhenium complexes. These species were further stabilized by the formation of transient host-guest aggregates with gamma cyclodextrin and finally converted into (188)Re(V)-DMSA through simple replacement of the coordinated ligands by H(2)DMSA. PMID- 10832089 TI - Mutagenic activity of a fluorinated analog of the beta-adrenoceptor ligand carazolol in the Ames test. AB - S-1'[(18)F]-Fluorocarazolol (FCAR) is a fluorinated analog of the nonmutagenic beta-blocker carazolol (CAR). In former studies FCAR proved to be suitable for quantification of beta-adrenoceptors in vivo with positron emission tomography (PET). We report here that FCAR displays no acute toxicity in either rats or mice. However, FCAR induces a strong dose-related increase in the number of revertants in the Ames test. We conclude that FCAR yields mutagenic activity as measured by the Ames test. PMID- 10832090 TI - Physiology and pathophysiology of sphingolipid metabolism and signaling. PMID- 10832091 TI - Acyl-CoA dependent acylation of phospholipids in the chloroplast envelope. AB - Acyl-CoAs are substrates for acyl lipid synthesis in the endoplasmic reticulum. In addition, they may also be substrates for lipid acylation in other membranes. In order to assess whether lipid acylation may have a role in plastid lipid metabolism, we have studied the incorporation of radiolabelled fatty acids from acyl-CoAs into lipids in isolated, intact pea chloroplasts. The labelled lipids were phosphatidylcholine (PC), phosphatidylglycerol (PG), phosphatidylinositol and free fatty acids. With oleoyl-CoA, the fatty acid was incorporated preferably into the sn-2 position of PC and the acylation activity mainly occurred in fractions enriched in inner chloroplast envelope. Added lysoPC stimulated the activity. With palmitoyl-CoA, the fatty acid was incorporated primarily into the sn-1 position of PG and the reaction occurred at the surface of the chloroplasts. As chloroplast-synthesized PG generally contains 16C fatty acids in the sn-2 position, we propose that the acylation of PG studied represents activities present in a domain of the endoplasmic reticulum or an endoplasmic reticulum derived fraction that is associated with chloroplasts and maintains this association during isolation. This domain or fraction contains a discreet population of lipid metabolizing activities, different from that of bulk endoplasmic reticulum, as shown by that with isolated endoplasmic reticulum, acyl CoAs strongly labelled phosphatidic acid and phosphatidylethanolamine, lipids that were never labelled in the isolated chloroplasts. PMID- 10832092 TI - Reverse hydrolysis reaction of a recombinant alkaline ceramidase of Pseudomonas aeruginosa. AB - Recently, we purified an alkaline ceramidase (CDase) of Pseudomonas aeruginosa and found that the enzyme catalyzed a reversible reaction in which the N-acyl linkage of ceramide was hydrolyzed or synthesized [J. Biol. Chem. 273 (1998) 14368-14373]. Here, we report the characterization of the reverse hydrolysis reaction of the CDase using a recombinant enzyme. The reverse hydrolysis reaction of the CDase was clearly distinguishable from the reaction of an acyl-coenzyme A (CoA) dependent N-acyltransferase, because the CDase catalyzed the condensation of a free fatty acid to sphingosine (Sph) without cofactors but did not catalyze the transfer of a fatty acid from acyl-CoA to Sph. The reverse hydrolysis reaction proceeded most efficiently in the presence of 0.05% Triton X-100 at neutral pH, while the hydrolysis reaction tended to be favored with an increase in the concentration of the detergent at alkaline pH. The specificity of the reverse reaction for fatty acids is quite broad; saturated and unsaturated fatty acids were efficiently condensed to Sph. In contrast, the stereo-specificity of the reverse reaction for the sphingoid bases is very strict; the D-erythro form of Sph, not the L-erythro or D/L-threo one, was only acceptable for the reverse reaction. Chemical modification of the enzyme protein affected or did not affect both the hydrolysis and reverse reactions to the same extent, suggesting that the two reactions are catalyzed at the same catalytic domain. PMID- 10832093 TI - Long-chain acyl-CoA dehydrogenase is a key enzyme in the mitochondrial beta oxidation of unsaturated fatty acids. AB - The first reaction of mitochondrial beta-oxidation, which is catalyzed by acyl CoA dehydrogenases, was studied with unsaturated fatty acids that have a double bond either at the 4,5 or 5,6 position. The CoA thioesters of docosahexaenoic acid, arachidonic acid, 4,7,10-cis-hexadecatrienoic acid, 5-cis-tetradecenoic acid, and 4-cis-decenoic acid were effectively dehydrogenated by both rat and human long-chain acyl-CoA dehydrogenases (LCAD), whereas they were poor substrates of very long-chain acyl-CoA dehydrogenases (VLCAD). VLCAD, however, was active with CoA derivatives of long-chain saturated fatty acids or unsaturated fatty acids that have double bonds further removed from the thioester function. Although bovine LCAD effectively dehydrogenated 5-cis-tetradecenoyl-CoA (14:1) and 4,7,10-cis-hexadecatrienoyl-CoA, it was nearly inactive toward the other unsaturated substrates. The catalytic efficiency of rat VLCAD with 14:1 as substrate was only 4% of the efficiency determined with tetradecanoyl-CoA, whereas LCAD acted equally well on both substrates. The conclusion of this study is that LCAD serves an important, if not essential function in the beta-oxidation of unsaturated fatty acids. PMID- 10832094 TI - Lipoprotein-associated alpha-tocopheryl-succinate inhibits cell growth and induces apoptosis in human MCF-7 and HBL-100 breast cancer cells. AB - alpha-Tocopheryl succinate (alpha-TS) is a potent inhibitor of tumor cell proliferation. The goal of the present study was to investigate whether and to what extent alpha-TS associates with plasma lipoproteins and if alpha-TS-enriched lipoproteins inhibit breast cancer cell growth in a manner comparable to the free drug. In vitro enrichment of human plasma revealed that alpha-TS readily associated with the main lipoprotein classes, findings confirmed in vivo in mice. At the highest alpha-TS concentrations, lipoproteins carrying 50000 (VLDL), 5000 (LDL) and 700 (HDL) alpha-TS molecules per lipoprotein particle were generated. alpha-TS enrichment generated lipoprotein particles with slightly decreased density and increased particle radius. To study whether the level of LDL-receptor (LDL-R) expression affects alpha-TS uptake from apoB/E containing lipoprotein particles human breast cancer cells with low (MCF-7) and normal (HBL-100) LDL-R expression were used. The uptake of free, VLDL- and (apoE-free) HDL(3)-associated alpha-TS was nearly identical for both cell lines. In contrast, uptake of LDL associated alpha-TS by HBL-100 cells (normal LDL-R expression) was about twice as high as compared to MCF-7 cells (low LDL-R expression). VLDL and LDL-associated alpha-TS inhibited proliferation most effectively at the highest concentration of alpha-TS used (100% inhibition of MCF-7 growth with 20 microg/ml of lipoprotein associated alpha-TS). However, also alpha-TS-free VLDL and LDL inhibited HBL-100 cell proliferation up to 55%. In both cell lines, alpha-TS-enriched HDL(3) inhibited cell growth by 40-60%. Incubation of both cell lines in the presence of free or lipoprotein-associated alpha-TS resulted in DNA fragmentation indicative of apoptosis. Collectively, the present findings demonstrate that: (1) alpha-TS readily associates with lipoproteins in vitro and in vivo; (2) the lipoprotein enrichment efficacy was dependent on the particle size and/or the triglyceride content of the lipoprotein; (3) uptake of LDL-associated alpha-TS was apparently dependent on the level of LDL-R expression; and (4) lipoproteins were efficient alpha-TS carriers inducing reduced cell proliferation rates and apoptosis in human breast cancer cells as observed for the free drug. PMID- 10832095 TI - Mass spectrometry monitoring of rhamnolipids from a growing culture of Pseudomonas aeruginosa strain 57RP. AB - Two rapid and simple methods for the characterisation and quantification of rhamnolipids produced by a growing culture of the Pseudomonas aeruginosa strain 57RP were developed. Two rhamnolipids were purified and their response factors determined. The various rhamnolipids produced were then measured using liquid chromatography/mass spectrometry. The culture supernatants were injected directly, without prior purification, in a HPLC equipped with a C(18) reverse phase column. The complete profile of rhamnolipid congeners produced during a 2 week cultivation period was monitored. In order to shorten the analysis time, another method was developed which did not require chromatographic separation of the rhamnolipids prior to their detection. Quantification of rhamnolipids using the direct infusion method gave results very similar to those obtained with HPLC separation. These two methods were very well correlated with the standard colorimetric orcinol method. The rhamnolipid profiles obtained show that the various rhamnolipid congeners are secreted simultaneously, and that their relative proportion remained unchanged throughout the cultivation period. PMID- 10832096 TI - Activation of astroglial phospholipase D activity by phorbol ester involves ARF and Rho proteins. AB - Primary cultures of rat cortical astrocytes express phospholipase D (PLD) isoforms 1 and 2 as determined by RT-PCR and Western blot. Basal PLD activity was strongly (10-fold) increased by 4beta-phorbol-12beta,13alpha-dibutyrate (PDB) (EC(50): 56 nM), an effect which was inhibited by Ro 31-8220 (0.1-1 microM), an inhibitor of protein kinase C (PKC), and by brefeldin A (10-100 microg/ml), an inhibitor of ADP-ribosylating factor (ARF) activation. Pretreatment of the cultures with Clostridium difficile toxin B-10463 (0.1-1 ng/ml), which inactivates small G proteins of the Rho family, led to a breakdown of the astroglial cytoskeleton; concomitantly, PLD activation by PDB was reduced by up to 50%. In contrast, inactivation of proteins of the Ras family by Clostridium sordellii lethal toxin 1522 did not affect PLD activation. In parallel experiments, serum-induced PLD activation was sensitive to brefeldin A, but not to Ro 31-8220 and not to clostridial toxins. We conclude that, in astrocytes, the PLD isoform which is activated by phorbol ester requires PKC, ARF and Rho proteins for full activity and probably represents PLD1. PMID- 10832098 TI - Kinetics and plasma concentrations of 26-hydroxycholesterol in baboons. AB - 26-Hydroxycholesterol (26OHC), a major oxysterol in human blood, is believed to play an important role in reverse cholesterol transport, bile acid formation, and regulation of various cellular processes. Using isotope dilution mass spectrometry, we measured plasma 26OHC concentrations in baboons fed either a high cholesterol/saturated fat (HC-SF) or normal chow diet. Plasma 26OHC levels in baboons were comparable to those reported for humans and were positively correlated with plasma cholesterol concentrations. Animals on the HC-SF diet had significantly higher 26OHC levels (0.274+/-0.058 microM, mean+/-S.D.) than those on the chow diet (0.156+/-0.046 microM). In separate experiments, [(3)H]26OHC was injected into four tethered baboons, and multiple blood samples drawn over a 1-h period were analyzed for [(3)H]26OHC and 26OHC. Fitting the specific radioactivity data to a two-pool compartmental model indicated a rapidly turning over plasma compartment (t(1/2) 2.9-6.0 min) and a second compartment with slow turnover (t(1/2) 76-333 min). The calculated 26OHC production rate was 2.5 micromol/kg body weight/day. Assuming all 26OHC is converted to bile acids, the 26OHC production rate corresponds to about 10% of total bile acid production in adult baboons. These results indicate that rapid turnover of plasma 26OHC at submicromolar concentrations could significantly contribute to bile acid synthesis. PMID- 10832097 TI - Phosphatidylinositol 3-kinase in zymosan- and bacteria-induced signalling to mobilisation of arachidonic acid in macrophages. AB - Stimulation of mouse peritoneal macrophages with zymosan or bacteria results in activation of 85-kDa cytosolic phospholipase A(2) (cPLA(2)) and release of arachidonate. We have investigated the role of phosphatidylinositol 3-kinase (PtdIns 3-kinase) in the signalling leading to activation of cPLA(2) and release of arachidonate in response to zymosan and the bacterium Prevotella intermedia. The specific PtdIns 3-kinase inhibitor wortmannin completely inhibited zymosan- and bacteria-induced release of arachidonate with an IC(50) value of 10-20 nM. Wortmannin also completely inhibited the zymosan-induced activation of cPLA(2), while the cPLA(2) activation by bacteria was partially inhibited by about 50%. Further experiments showed that zymosan-induced activation of extracellular signal-regulated kinase was inhibited, and bacteria-induced activation of the kinase strongly reduced, in the presence of wortmannin. Also zymosan-induced activation of p38 mitogen-activated protein kinase was inhibited by wortmannin, while p38 activation induced by bacteria was not. The zymosan- and bacteria induced activation of phospholipase C, as determined by the generation of inositol phosphates, was also inhibited by wortmannin. Moreover, zymosan caused activation of PtdIns 3-kinase, which was totally inhibited by wortmannin. In contrast to zymosan and bacteria, arachidonate release induced by calcium ionophore alone, or further amplified by phorbol ester, was not sensitive to wortmannin. These results suggest that PtdIns 3-kinase constitutes a critical component in the zymosan- and bacteria-induced signalling leading to release of arachidonate and that PtdIns 3-kinase is positioned upstream of phospholipase C in this pathway. PMID- 10832099 TI - Differential effects of acyl-CoA binding protein on enzymatic and non-enzymatic thioacylation of protein and peptide substrates. AB - Both enzymatic and autocatalytic mechanisms have been proposed to account for protein thioacylation (commonly known as palmitoylation). Acyl-CoA binding proteins (ACBP) strongly suppress non-enzymatic thioacylation of cysteinyl containing peptides by long-chain acyl-CoAs. At physiological concentrations of ACBP, acyl-CoAs, and membrane lipids, the rate of spontaneous acylation is expected to be too slow to contribute significantly to thioacylation of signaling proteins in mammalian cells (Leventis et al., Biochemistry 36 (1997) 5546-5553). Here we characterized the effects of ACBP on enzymatic thioacylation. A protein S acyltransferase activity previously characterized using G-protein alpha-subunits as a substrate (Dunphy et al., J. Biol. Chem., 271 (1996) 7154-7159), was capable of thioacylating short lipid-modified cysteinyl-containing peptides. The minimum requirements for substrate recognition were a free cysteine thiol adjacent to a hydrophobic lipid anchor, either myristate or farnesyl isoprenoid. PAT activity displayed specificity for the acyl donor, efficiently utilizing long-chain acyl CoAs, but not free fatty acid or S-palmitoyl-N-acetylcysteamine. ACBP only modestly inhibited enzymatic thioacylation of a myristoylated peptide or G protein alpha-subunits under conditions where non-enzymatic thioacylation was reduced to background. Thus, protein S-acyltransferase remains active in the presence of physiological concentrations of ACBP and acyl-CoA in vitro and is likely to represent the predominant mechanism of thioacylation in vivo. PMID- 10832100 TI - Apolipoprotein-mediated cellular cholesterol/phospholipid efflux and plasma high density lipoprotein level in mice. AB - Helical apolipoprotein(apo)s generate pre-beta-high density lipoprotein (HDL) by removing cellular cholesterol and phospholipid upon the interaction with cells. To investigate its physiological relevance, we studied the effect of an in vitro inhibitor of this reaction, probucol, in mice on the cell-apo interaction and plasma HDL levels. Plasma HDL severely dropped in a few days with probucol containing chow while low density protein decreased more mildly over a few weeks. The peritoneal macrophages were assayed for apoA-I binding, apoA-I-mediated release of cellular cholesterol and phospholipid and the reduction by apoA-I of the ACAT-available intracellular cholesterol pool. All of these parameters were strongly suppressed in the probucol-fed mice. In contrast, the mRNA levels of the potential regulatory proteins of the HDL level such as apoA-I, apoE, LCAT, PLTP, SRB1 and ABC1 did not change with probucol. The fractional clearance rate of plasma HDL-cholesteryl ester was uninfluenced by probucol, but that of the HDL apoprotein was slightly increased. No measurable CETP activity was detected either in the control or probucol-fed mice plasma. The change in these functional parameters is consistent with that observed in the Tangier disease patients. We thus concluded that generation of HDL by apo-cell interaction is a major source of plasma HDL in mice. PMID- 10832101 TI - Modifications of glycosphingolipid profile and synthesis in normal rat fibroblasts and in syngeneic neoplastic cells at different subculture stages. AB - Glycosphingolipids are plasma membrane macromolecules involved in diversified recognition functions on the cell surface resulting in modulation of cell adhesion and differentiation. As the in vitro cellular system of the neoplastic cell line SGS/4A and syngeneic normal fibroblasts (FG) represents a useful tool for studies on molecular mechanisms regulating cell adhesion, neoplastic transformation and cellular ageing, we studied the changes of glycosphingolipid and of the enzymes involved in their metabolism in both cultured cells at different subculture stages. The FG subculture progression induces a drastic decrease of total glycosphingolipid content with consistent alterations in the molecular composition. In particular, a significant decrease of GM(3), a slight increase of GD(1a), the disappearance of 'b'-series gangliosides and the drastic reduction of triosylceramides were observed. On the contrary, the increasing number of SGS/4A subcultures, characterized by a specific and different glycosphingolipid composition as compared with FG cells, does not cause modifications. Although glycosyltransferase activity levels quite well parallel the glycosphingolipid patterns and can account for the noted variations, the mRNA expression analysis of two glycosyltransferases suggests that the in vitro cell ageing of normal rat fibroblasts causes drastic changes in the glycosphingolipid profile through the regulation, at either the transcriptional or post translational level, of some biosynthetic enzymes. PMID- 10832102 TI - Isolevuglandin-protein adducts in humans: products of free radical-induced lipid oxidation through the isoprostane pathway. AB - A family of extremely reactive electrophiles, isolevuglandins (isoLGs), is generated in vivo by free radical-induced lipid oxidation and rearrangement of endoperoxide intermediates of the isoprostane pathway. Protein adducts of two different oxidized lipids, isoLGE(2) and iso[4]LGE(2), and the corresponding autoantibodies are present in human blood. Western blot analysis of a polyacrylamide gel electrophoresis gel detects several immunoreactive plasma proteins. Only a minor fraction of the isoLG-protein modifications is associated with low density lipoprotein since mean levels were decreased only 20-22% by immunoprecipitation of apolipoprotein B (apoB). Mean levels of both isoLGE(2) and iso[4]LGE(2)-protein adducts in plasma from patients with atherosclerosis (AS) (n=16) or end-stage renal disease (RD) (n=8) are about twice those in healthy individuals (n=25). These elevated levels are not related to variations in age, total cholesterol or apoB. A linear correlation (r=0.79) between plasma isoLGE(2) and iso[4]LGE(2)-protein adduct levels in all 49 individuals is consistent with a common free radical-induced mechanism for the production of both oxidized lipids in vivo. The correlation is even stronger (r=0.86) for patients with AS or RD. That isoLG-protein adduct levels are more strongly correlated with disease than are total cholesterol or apoB suggests an independent defect that results in an abnormally high level of oxidative injury associated with AS and RD. PMID- 10832103 TI - Cloning and expression of rat neutral sphingomyelinase: enzymological characterization and identification of essential histidine residues. AB - Using cross-species sequence homology, we cloned a cDNA for rat neutral sphingomyelinase (nSMase) composed of 422 amino acids that shares 87.6 and 79.0% identity with the mouse and human forms respectively. The rat nSMase expressed in Escherichia coli catalyzed sphingomyelin hydrolysis at neutral pH in a Mg(2+) dependent manner, and required Triton X-100, dithiothreitol, and KCl for its full activity. The cloned rat enzyme shares conserved sequences with nSMases from both eukaryotes and prokaryotes. Introduction of single mutations into either of the histidine residues at positions 136 and 272, putative active sites, entirely abolished the activity, supporting a common mechanism for the nSMase family independent of the species. However, mutation in histidine 151, conserved only in eukaryotes, also abolished the activity, suggesting eukaryote-specific control of nSMase linked to this histidine 151. This enzyme also catalyzed the hydrolysis of lyso-platelet activating factor to yield 1-alkylglycerol at a rate that is slightly lower than that with sphingomyelin. PMID- 10832104 TI - Graves' ophthalmopathy: eye muscle involvement in patients with diplopia. AB - BACKGROUND: Diplopia identifies patients with eye muscle involvement in Graves' ophthalmopathy (GO). OBJECTIVE: To identify clinical parameters that could eliminate the need for magnetic resonance imaging (MRI) to assess the activity of inflammation in the eye muscles of GO patients with diplopia. METHODS: In 43 patients with GO with recently developed diplopia, orbital ultrasound and MRI were performed. Muscle diameters and MRI T2 relaxation times were measured, and the amount of orbital connective tissue was calculated from MRI scans and compared with ultrasound readings, diplopia grades, degree of protrusion, ocular pressure, tear production, antibody levels and hormonal parameters of thyroid function. RESULTS: No correlation was found between diameters of 233 extraocular muscles measured by MRI and by ultrasound. For each of the four muscles, there was a diameter above which ultrasound was always unreliable. MRI data were used in further analysis. Of the muscles examined, the inferior rectuses were the most frequently enlarged - at least one, in 93% of cases. Medial, lateral and superior rectuses were enlarged in 59%, 37% and 34% of the orbits respectively. The pattern of muscle involvement of the two orbits tended to be symmetric (r=0.49, P=0.003), particularly for the medial rectuses (r=0.90, P=0.000). Proptosis correlated with the sum of the muscle diameters for a given eye (right eye: r=0.54, P=0.003; left eye: r=0.57, P=0.001), but it failed to correlate with the amount of orbital connective tissue. In 53% of the patients, normal T2 relaxation times were found in all eight muscles. There was only a weak correlation between muscle thickness and T2 relaxation time (r=0.49, P=0.003), indicating that muscle enlargement alone is not a sign of disease activity. The severity of diplopia was independent of T2 relaxation time. The amount of orbital connective tissue showed a negative correlation with the greatest T2 relaxation time for a given eye (r= 0.52, P=0.004); this suggests that disease types exist that have predominant muscle involvement and predominant connective tissue expansion. No correlation between connective tissue expansion and proptosis, diplopia grade, muscle thickness or disease duration was found - that is, connective tissue expansion is not a major factor in diplopia. Both muscle and connective tissue findings were independent of thyroid function. CONCLUSION: Ultrasound and MRI eye muscle diameter readings do not correlate, because of the inherent inaccuracy of orbital ultrasound. Muscle enlargement alone does not mean oedematous swelling and active disease. Neither ultrasound, nor any combination of 11 clinical and laboratory parameters provided the degree of information on muscles and connective tissue that was obtainable by MRI. In unclear cases of recently developed diplopia, before orbital decompression surgery, in the case of treatment failure or if, for any other reason, imaging is needed in GO, MRI is the method of choice. PMID- 10832105 TI - LHRH might act as a negative autocrine regulator of proliferation of human ovarian cancer. AB - OBJECTIVE: More than 80% of human ovarian cancers express LHRH and its receptor. The proliferation of human ovarian cancer cell lines is reduced by both LHRH agonists and antagonists. This study was designed to further clarify the possible biological function of this LHRH system. DESIGN: As LHRH agonists and antagonists uniformly reduce proliferation of human ovarian cancer in a dose-dependent way, the effect of low concentrations of authentic LHRH was studied. In addition, longer periods of treatment (up to 9 days) were analyzed. To assess the physiological role of LHRH produced by ovarian cancer cells it was neutralized by adequate concentrations of a specific LHRH antiserum. METHODS: Human ovarian cancer cells EFO-21 and EFO-27, which express LHRH and its receptor, were incubated for 1-9 days with increasing concentrations (1pmol/l to 10 micromol/l) of authentic LHRH or with concentrations of LHRH antiserum capable of neutralizing at least 1nmol/l LHRH. Proliferation was assessed by counting cells. RESULTS AND CONCLUSIONS: Authentic LHRH reduced time- and dose-dependently proliferation (by maximally mean+/-s.e.m. 32.7 +/- 4.4%, Newman-Keuls, P < 0.001) of both ovarian cancer cell lines. At very low concentrations (1pmol/l) a marginal reduction of proliferation or no effect was observed. A mitogenic effect of authentic LHRH was never detected. Treatment of ovarian cancer cell cultures with antiserum to LHRH significantly increased (up to mean+/-s.e.m. 121.0 +/- 2.8% of controls, Newman-Keuls P <0.001) proliferation of EFO-21 and EFO-27 cells. These findings suggest that LHRH produced by human ovarian cancer cells might act as a negative autocrine regulator of proliferation. PMID- 10832106 TI - Expression of low and high density lipoprotein receptor genes in human adrenals. AB - Corticosteroids are synthesized from cholesterol which may arise from de novo synthesis or from the uptake of low or high density lipoproteins (LDL or HDL). In the present study, we compared the expression and regulation patterns of LDL receptor and CLA-1 (CD36 and LIMPII Analogous-1, an HDL receptor) genes in adult human adrenocortical tissues to shed more light on the relative contribution of LDL and HDL in human adrenal steroidogenesis. By screening 64 normal and pathological adrenal samples by Northern blotting, we found a positive correlation between LDL receptor and CLA-1 mRNA expression in the adrenal tissues (r=0.547; spearman rank correlation test P<0.01). Adrenal tissues adjacent to Cushing's adenomas contained consistently less LDL receptor and CLA-1 mRNA than normal adrenals (Mann-Whitney P<0.05). In primary cultures of normal adrenal cells, accumulation of both LDL receptor and CLA-1 mRNAs was upregulated by ACTH in a dose- and time-dependent manner, with an earlier induction of LDL receptor than CLA-1 mRNA expression. (Bu)(2)cAMP also increased the levels of these two mRNAs. Addition of LDL, but not HDL, into the culture medium increased cortisol production in untreated adrenocortical cells. Both LDL and HDL enhanced ACTH induced cortisol production, with the effect of LDL much stronger than that of HDL. Our data show that LDL receptor and CLA-1's expression is ACTH-dependent and occurs in parallel in human adrenal tissues. LDL rather than HDL may be used as the preferential source of cholesterol for steroidogenesis in human adult adrenocortical cells. PMID- 10832107 TI - Thyroid cancer in the new millennium PMID- 10832108 TI - Evaluation of the thyroid nodule. AB - BACKGROUND: Clinically detectable thyroid nodules occur in up to 4% of the population in the United States. With ultrasound, nodules may be found in up to 50% of those over 50 years of age. METHODS: The author reviews his own experience as well as that of others to define a sound clinical approach to the differential diagnosis and detection of thyroid cancer. RESULTS: Prior neck irradiation is a risk factor for thyroid malignancy. The association of a thyroid nodule with enlarged lymph nodes or fixation of the nodule to strap muscles or the trachea suggests malignancy. A diffusely multinodular gland is usually benign. CONCLUSIONS: Thyroid function tests rarely help a differential diagnosis. Fine needle aspiration is the "gold standard" for diagnosis. Tiny "incidentalomas" are often followed with repeat monitoring for change of size or character. PMID- 10832109 TI - Prognostic indicators in differentiated thyroid carcinoma. AB - BACKGROUND: Thyroid cancer ranges from well-differentiated lesions with an excellent prognosis to anaplastic carcinoma, which is almost uniformly fatal. Thus, methods to assess the behavior of thyroid malignancies are necessary to arrive at appropriate treatment decisions. METHODS: We discuss the factors that affect the prognosis of patients with well-differentiated thyroid malignancies, including papillary, follicular, Hurthle cell, and medullary thyroid carcinomas. We also review the presentation, therapy, and outcome of patients seen at our center over a span of 50 years. These data have identified those prognostic factors that are predictive of survival and recurrence in differentiated thyroid cancer. RESULTS: Several classifications with different variables have been developed to define risk-group categories. Three widely used systems, in addition to the TNM staging system, include AGES, AMES, and MACIS. CONCLUSIONS: A better understanding of independently important prognostic variables will result in improved patient care and treatment. PMID- 10832110 TI - Thyroid cancer: extent of thyroidectomy. AB - BACKGROUND: Surgical resection is the key to management of thyroid cancer, but determining the optimal surgical procedure for individual cases has been controversial. METHODS: The author reviews several large data bases that allow examination of prognostic criteria for long-term outcomes. RESULTS: Patients can be classified into good- or poor-risk groups that assist defining the optimal surgical procedure. Routine use of total thyroidectomy in all patients with thyroid cancer is best avoided; however, patients with medullary cancer generally need total thyroidectomy. CONCLUSIONS: The definition of risk groups has clarified the options regarding choice of primary surgical therapy for differentiated thyroid cancer. PMID- 10832111 TI - Evaluation and treatment of aerodigestive tract invasion by well-differentiated thyroid carcinoma. AB - BACKGROUND: Although rare, invasion of the upper aerodigestive tract by well differentiated thyroid carcinoma can be a source of significant morbidity as well as mortality for the patient. Effective management of patients with invasive thyroid carcinoma requires an understanding of the patterns of invasion and methods of treatment, including surgical resection and adjuvant therapy. METHODS: The author reports on experience with invasive well-differentiated thyroid carcinoma, discussing diagnosis (based on physical examination, imaging studies, and endoscopy) as well as treatment options (based on degree of aerodigestive tract invasion). RESULTS: Direct intraluminal invasion of thyroid carcinoma requires definitive resection of aerodigestive tract lumen to remove all gross disease. However, when the lumen is not involved, "shaving" tumor from airway or esophagus is an acceptable treatment with a similar locoregional control rate and minimal morbidity when compared to definitive aerodigestive tract resection. CONCLUSIONS: Successful treatment of invasive thyroid carcinoma should improve survival and reduce not only the morbidity of the disease, but also the morbidity of the surgical procedure. PMID- 10832113 TI - Evidence-based management of Hodgkin's disease: the role of autologous stem cell transplantation. PMID- 10832112 TI - Medullary carcinoma of the thyroid. AB - Medullary thyroid cancer (MTC) is a distinct C-cell tumor of the thyroid. We review the oncogenesis and management of both sporadic tumors and those tumors arising as part of specific inherited syndromes. The RET proto-oncogene plays a role in the development of inherited forms of MTC and has become important in the clinical management of patients and their families. The recognition of the high rate of regional nodal involvement has led to lymphadenectomy being strongly considered for patients undergoing thyroidectomy for MTC. PMID- 10832114 TI - Tumors of the thyroid gland: histologic and cytologic features--part 1. PMID- 10832115 TI - Nutritional factors and endometrial cancer in Ontario, Canada. PMID- 10832116 TI - [The cardiology community and health reform. Some reflections to open a debate]. AB - A recently introduced health care reform in Italy will modify substantially the scenario in which all physicians and also the cardiac specialists working in and out of the hospitals will operate. Therefore it is important that the cardiological community, who manages a large proportion of human and financial resources of the Italian National Health Service, knows the reform and interacts with the national and local authorities for the best implementation of the basic principles on which the reform has been founded. These principles are the following: the Italian health service will guarantee all citizens the so-called "essential levels of care" identified in accordance with four distinctive features. They must: 1) safeguard real needs of care (i.e. cosmetic surgery is not considered), 2) be evidence based, 3) be appropriate for individual patients, 4) be cost-effective. In a context of scarce resources and rapidly increasing demand of care this basic strategy seems to be the only one suitable for a National Health Service, but the application of this principle in the real world of care seems a very difficult task, and the role of medical associations is obviously crucial for a good outcome. This report illustrates some articles of the law that deal with the medical profession: guidelines and appropriateness of the criteria; accreditation, clinical competence and quality control; continuing medical education. PMID- 10832117 TI - [The chain of survival]. AB - Coronary artery disease remains the leading cause of death in Italy as in most developed countries. Many of the victims die from sudden cardiac arrests, most commonly resulting from out-of-hospital ventricular fibrillation. The epidemiological evidence of the high incidence of sudden deaths in the small subgroup of high-risk patients and of the low incidence of events in the large subgroup of low- and intermediate-risk patients which are contributing to most of the deaths has suggested the development of emergency services and to the chain of survival concept. With the development of the automatic external defibrillator, early recognition and treatment of ventricular fibrillation by non skilled rescuers has significantly improved the outcome of sudden death. Training is essential to provide the numerous skills required for those regularly exposed to resuscitation and also to provide the diffusion of cardiopulmonary resuscitation competence in the general population. This paper reviews the past, present and future development of the epidemiological, organizational, training, and research issues related to the chain of survival. PMID- 10832118 TI - [The treatment and outlook of atherosclerosis today]. AB - Nowadays, the success of atherosclerosis therapy and the efficacy of cardiovascular disease prevention can rely on a comprehensive strategy. New algorithms derived from large population studies have improved the risk evaluation for the single individual and, consequently, the therapeutic approach can be better adapted to the specific needs of the patient. Improvements of noninvasive techniques, such as B-mode ultrasound and magnetic resonance imaging, allow for the reproducible and precise monitoring of lesion evolution that can be easily applied to large population samples. Ongoing studies regarding the relationship between circulating markers of atherosclerosis and the occurrence of cardiovascular events will improve our assessment of impending risk. Finally, the presence of pleiotropic effects of statins in vivo (and, perhaps, of fibrates as well) underscores the added value of this class of drugs in the treatment of lipid disorders and in the prevention of cardiovascular events. Prospectively, the integration of all these approaches will have a desirable impact on the qualitative improvement of human longevity. PMID- 10832119 TI - [Tissue Doppler: the physical principles, representational and analytical modalities and clinical applications]. AB - Tissue Doppler imaging or myocardial velocity imaging is a variation of conventional Doppler. This modality allows the quantification of the Doppler shift within the range of myocardial tissue motion. The velocity of motion at a variety of myocardial sites can be determined and distinguished very rapidly using Doppler techniques. The velocity of moving tissue can be studied with pulsed wave tissue Doppler sampling, which displays the velocity of a selected myocardial region against time, with high temporal resolution. In addition, the velocities can be calculated with time velocity maps and displayed as color coded velocity maps in either M-mode or two-dimensional format. This review will focus on the technical aspects and the different methods of tissue Doppler for regional systolic and diastolic left ventricular function analysis. While pulsed wave tissue Doppler allows us to measure the velocities of a selected myocardial region, color tissue Doppler gives the best overall view of cardiac dynamics because the whole scanned color data are displayed simultaneously. However, there is an increasing need for objective evaluation of tissue Doppler information. Digital images and data post-processing allow for quantitative off-line analysis, and the different approaches and parameters proposed from different centers are discussed. In recent years, tissue Doppler imaging has been applied for accurate evaluation of diastolic function, quantifying regional function particularly during stress, pre-excitation syndrome, and left ventricular hypertrophy. The results of these experiences indicate that tissue Doppler imaging is a promising technique for quantifying the response of the myocardium and endocardium during both normal and abnormal function. Again, there is a significant learning curve concerning its application, but with experience it will be a useful and reproducible technique. PMID- 10832120 TI - [The role of biochemical markers of myocardial damage in clinical practice: the diagnosis of infarct and risk stratification. The Intersociety Interdisciplinary Study Group of the ANMCO-SIBioC-SIMeL, Markers of Muocardial Lesions. L'Associazione Nazionale Medici Cardiologi Ospedalieri-Societa Italiana di Biochimica Clinica-Societa Italiana di Medicina di Laboratorio]. AB - For many years creatine kinase (CK) and CK-MB isoenzymes were used together with the ECG to confirm the presence of myocardial infarction. During the last decade newer cardiac markers have been introduced and immunological test systems developed for their quantification. Among these new markers, a prominent role has emerged for cardiac troponins (T or I). These technological advanced assays have shown greater sensitivity compared to "conventional cardiac enzymes;, thereby identifying patients with small--at times, microscopic--infarcts who would not have met defining criteria for myocardial infarction in an earlier era. Another major advantage shown by both cardiac troponins with respect to "conventional cardiac enzymes" is their ability to predict clinical outcome over a short- or long-term follow-up in patients with acute coronary syndromes, and this appears to be particularly relevant in patients with micronecrosis, who constitute a high risk subgroup of unstable angina patients. Recently, myoglobin has also been widely applied as a marker. Although lacking in myocardial specificity, it is the earliest marker to show an increase after coronary occlusion. Thus, the combined use of myoglobin and a cardiospecific structural protein such as troponin T or I may prove an attractive strategy for biochemical testing in chest pain patients. With the routine use of these novel cardiac markers, fascinating opportunities are now open in the field of diagnostic classification (making the World Health Organization definition of myocardial infarction obsolete) and risk stratification in chest pain patients; opportunities that were unforeseen in the era of cardiac enzymes. However, the use of these markers has also posed some important questions on: a) the best and most cost-effective diagnostic strategy in chest pain patients; b) the remaining role of cardiac enzymes; c) the therapeutic consequences of a positive test result. PMID- 10832121 TI - [Management control of cardiology: the experience of a departmental unit]. AB - In most Italian hospitals, sanitary reform is being applied, while at the same time a new organization of the National Health System is being planned. The director of the medical hospital (head doctor) is becoming more and more involved in management and this aspect has modified his professional attributes. Cardiology is a branch of medicine that, through its scientific preparatory work consisting in debates, management courses, ethics, and production of managerial software, is closer to applying the reform without risking improper administrative aspects. This, obviously, comes about after thoroughly reviewing past work methods and the need to have an administrative organization, which allocates efficient use of manpower and materials, helping to eliminate any sources of inefficiency. The logical procedure foresees an actual analysis in terms of sanitary needs and availability of resources, and so attempting to better balance and harmonize both aspects of the problem. Certainly, the acquisition of theoretical norms and practices, which today are present because of the upsurge in training courses for doctors, is not enough to guarantee the achievement of optimal results. Furthermore, we find that theoretical models need to be validated and adapted to real work situations in the public hospital sector. This paper proposes, therefore, to explain the managerial experiences achieved in actual work situations at the Cardiology Department Unit of the San Giovanni Addolorata Hospital in Rome. In particular, it shows that in order to reach its clinical and economical objectives, it is essential to make available correct informative support for strategic and operational decisions. We can observe that there is a continuing lack of computer support systems being integrated into the present organization of most cardiology units. The use of software distributed to cardiology units from the Associazione Nazionale Medici Cardiologi Ospedalieri (ANMCO) has enabled us to partially remedy the above mentioned inadequacy and, at the same time, has pointed out the limits of automated support with regards managerial needs which are rapidly changing and becoming increasingly more complex. The experience described shows the possibility of uniting an efficient organization, from a clinical point of view, with an adequate managerial automation without including any flaws from the previous organizational systems. Based on achieved results and the difficulties encountered, further development in computer technology support to hospital management is suggested. This paper, in particular, stresses the need to improve automated informative support available to doctors in managerial positions. This improvement would allow us to immediately evaluate "internal" administrative matters, have the data available for budget planning and analysis of budget variations, and be better able to integrate this with clinical data. The scientific community can give further assistance by promoting the genesis of data for a sensible bench-marking within medical organizations which is the only means sufficient to give reliable and clinically appropriate indications on the possibility of increasing its own structural efficiency. PMID- 10832122 TI - [The evaluation of the appropriateness of instrumental exams in cardiology. The case of echocardiography]. AB - BACKGROUND: The aim of this study was to evaluate the appropriateness and utility rates of echocardiograms performed in 309 patients in an outpatient clinical setting. METHODS: Data were collected by means of a questionnaire filled in by the cardiologists who performed the examinations. Appropriateness was evaluated according to international guidelines and scored as class I: appropriate, class II: doubtful appropriateness, class III: inappropriate; the exam was deemed useful if it was able to influence the clinical decision-making; normalcy rate was also checked. The relationship between both the referring physicians and motivation of the exam and its appropriateness, and the relationship between appropriateness and both the normalcy rate and utility of the exam were assessed. RESULTS: An echocardiogram was requested by the cardiologist in 46% of patients; the more common reasons for the exam were arterial hypertension (26%), cardiac murmur (18%), palpitations (15%), and known coronary artery disease (10%). The echocardiogram was appropriate (class I) in 25% of patients, doubtfully appropriate (class II) in 39% of patients and inappropriate (class III) in 36% of patients. The appropriateness rate between the cardiologists was similar to that of other prescribing clinicians (p = NS). The highest class III rate was found in patients with hypertension, while the highest class I rate was found in patients with a cardiac murmur (p < 0.01). Normalcy rate was lower in class I than in class II and III exams (p < 0.001). The utility rate was higher in class I (76%) than in class II (13%) and III (< 1%) exams (p < 0.01). CONCLUSIONS: International guidelines can be used effectively and safely to identify (not to prescribe) the useless echocardiograms. PMID- 10832123 TI - [The safety and efficacy of systemic salvage thrombolysis in acute myocardial infarct]. AB - BACKGROUND: Thrombolysis reduces mortality in patients with acute myocardial infarction hospitalized within 6 hours of the symptom onset. Infarctions involving a small area of the myocardium show a lower mortality in comparison to those involving a large area. The aim of this study was to evaluate the safety and efficacy of rescue thrombolysis in patients with large acute myocardial infarction who had failed standard thrombolysis. METHODS: From January 1995 to December 1997, ninety patients (69 males, 21 females, mean age 56.7 +/- 9 years), hospitalized for suspected acute myocardial infarction within 4 hours of the symptom onset, suitable for thrombolysis (first episode), and who experienced pain and showed persistent ST segment elevation 120 min after starting thrombolysis, were randomized (single blind) into two groups: Group A (n = 45) received an additional thrombolytic treatment (rt-PA 50 mg), 10 mg as a bolus plus 40 mg in 60 min; Group B (n = 45) received conventional therapy. Positive non-invasive markers were defined as follows: resolution of chest pain; > 50% reduction in ST segment elevation; double marker of creatine phosphokinase (CPK) and CK-MB activity 2 hours after the start of thrombolysis; occurrence of reperfusion arrhythmias within the first 120 min of thrombolytic therapy. Blood pressure, heart rate and ECG were continuously monitored. Echocardiogram was carried out at entry and before discharge to control ejection fraction and segmental wall motion. Adverse events such as death, reinfarction, recurrent angina, incidence of major and minor bleeding, and emergency bypass surgery or coronary angioplasty were checked. RESULTS: Thirty-five patients (77.7%) showed reperfusion (10-50 min) after the start of additional rt-PA. In patients who did not receive additional thrombolysis, only 12 (26.6%) showed reperfusion 65-115 min after the end of rt-PA infusion. Group A showed an earlier and lower CK and CK-MB peak than Group B (p = 0.0001, p = 0.009, and p = 0.002, respectively). Mortality (n = 16, 17.7%) was higher in Group B (n = 13) than in Group A (n = 3) (28.8 vs 6.6%, p = 0.041). Seven patients from Group A showed non-fatal reinfarction. Angina was observed in 18 (40%) patients from Group A and 3 (6.6%) from Group B (p = 0.006). Ten of these patients underwent urgent coronary angioplasty (9 from Group A and 1 from Group B) and 3 from Group A urgent bypass surgery. Minor bleeding was higher in Group A than in Group B (44.4 vs 15.5%, p = 0.047). A major bleeding was observed in Group A (non-fatal stroke). At predischarge echocardiogram ejection fraction was higher in Group A than in Group B (46 +/- 8 vs 38 +/- 7%, p = 0.0001). CONCLUSIONS: Our data suggest that an additional dose of a thrombolytic drug in patients with unsuccessful thrombolysis is feasible, and the bleeding increase is an acceptable risk in comparison with the advantages obtained from a reduced infarct extension. Rescue thrombolysis could save time and allow mechanical revascularization to be carried out in patients admitted to a hospital without interventional cardiology laboratory or in those who have to be refereed to other hospitals for urgent bypass surgery. PMID- 10832124 TI - [The initial experience of the use of ReoPro in high-risk coronary angioplasty in Italy: the Italian Registry on the Conditions for the Use of ReoPro during Angioplasty (R.I.CO.R.D.A.)]. AB - BACKGROUND: The R.I.CO.R.D.A. Registry was created with the aim of monitoring early Italian experience on the use of abciximab (ReoPro) in the setting of high risk coronary angioplasty (PTCA). Indication for enrollment included abciximab administration either on an elective basis (i.e. preprocedural or planned) or as bailout for complicated PTCA (i.e. intraprocedural, as a rescue treatment). METHODS: From June 1996 to January 1998, 359 patients undergoing PTCA on 443 lesions were enrolled in 24 Italian catheterization laboratories. In 91/359 patients (25%) abciximab was administered as bailout, and in the remaining cases on an elective basis as a pre-PTCA treatment. RESULTS: The incidence of death, coronary artery bypass grafting, Q wave and large non-Q wave myocardial infarction, small non-Q wave myocardial infarction and recurrent ischemia in the elective group were 1.9, 1.9, 2.6, 6.7 and 3.4%, respectively. In the bailout group corresponding figures were 4.4 (p = NS), 3.3 (p = NS), 16.5 (p < 0.01), 16.5 (p < 0.01) and 12.1% (p < 0.01), respectively. The heparin dose during PTCA was quite variable, particularly in the very early stages. Overall, 46% of patients received a heparin dose of < or = 5000 IU, and the remaining a higher dose, more often 10,000 IU. The latter group showed a significantly higher incidence of bleeding complications (25 vs 10%, p < 0.01). The incidence of bleeding was almost double in patients treated with bailout abciximab as compared with the elective group (29 vs 15%, p < 0.01), according to the higher heparin dose administered (9253 +/- 3341 vs 6649 +/- 3156 IU, p < 0.01). Overall, the incidence of red blood cell transfusions was 5.8%. CONCLUSIONS: The results of this Registry shed light on the actual use of abciximab in the setting of high risk PTCA in Italy. The incidence of cardiac events and bleeding complications in the group of patients treated on an elective basis is comparable to that reported in the main large-scale international trials. The bailout abciximab group showed a higher incidence of both cardiac events and bleeding complications. PMID- 10832125 TI - [Epidural spinal stimulation in the treatment of refractory angina pectoris. Its clinical efficacy, complications and long-term mortality. An Italian multicenter retrospective study]. AB - BACKGROUND: Spinal cord stimulation has been used for many years in the treatment of refractory angina pectoris. Its anti-anginal and anti-ischemic effect has been well documented in several studies, but the long-term efficacy, safety and survival rate are not well known. The aim of this study was to carry out a retrospective analysis of a series of patients from the Italian Multicenter Registry, the data of which were collected in five centers, by means of a questionnaire. METHODS: One hundred and thirty patients (83 males, 47 females, mean age 74.8 +/- 9.8 years) were submitted to spinal cord stimulator implantation for refractory angina pectoris in the period 1988-1995 and controlled during a mean follow-up of 31.4 +/- 25.9 months. A previous myocardial infarction had already occurred in 69.3% of patients, whereas in 67.6% multivessel coronary artery disease was documented. A left ventricular dysfunction (ejection fraction < 0.40) was present in 34% of patients; bypass surgery and coronary angioplasty were performed in 49.6% and in 27% of patients respectively. In 96.3% of cases revascularization procedures were not advisable. RESULTS: A complete follow-up of 116 patients (89.2%) was available. The spinal cord stimulator induced a significant reduction in NYHA functional class from 2.5 +/- 1.2 to 1.5 +/- 0.9 (p < 0.01). During the follow-up 41 patients (35.3%) died, and in 14.2% a new acute myocardial infarction developed. The total percentage of minor spinal cord stimulation-related complications was 6.8%. No major complications occurred. The annual total mortality rate was 6.5%, whereas the cardiac mortality rate was 5%. Compared to the survivors, patients who died showed a higher incidence of left ventricular dysfunction, previous myocardial infarction and bypass surgery at implantation. CONCLUSIONS: In our experience, spinal cord stimulation is an effective therapy in patients affected by refractory angina pectoris and who cannot undergo revascularization procedure. The complication rate is low, with the total and cardiac mortality showing a trend as that reported for patients with similar coronary disease. PMID- 10832126 TI - [The correlation between the type of positivity of the tilt test and a simultaneous electroencephalogram: the preliminary results]. AB - BACKGROUND: Today the first-choice examination to study neurally-mediated syncope is the tilt test. There are still many aspects to be clarified on the pathophysiology of neurally-mediated syncope, and much uncertainty remains on the therapeutic procedure to adopt. Recent research has investigated the role of neurohumoral agents, thus shifting interest to the pathogenetic role of the central nervous system, over and above that of the already widely studied vegetative nervous system. This is why we decided to carry out the tilt test with simultaneous electroencephalogram (EEG) recordings, with the aim of documenting any possible correlation between test positivity, according to Sutton's classification, and the EEG results. METHODS: We studied 15 patients (8 males, 7 females, age range 18-74 years) with a history of repeated syncopal and presyncopal episodes who had formerly undergone numerous clinical and instrumental examinations, including EEG, with negative results. The tilt test was carried out with continuous measurement of blood pressure (Ohmeda Finapres System) and simultaneous EEG recording. RESULTS: Ten patients (66%) were positive, 6 had experienced syncope episodes (4 type 2A and 2 type 1) and 4 presyncope (1 type 2A and 3 type 1). In all the syncope positive patients the EEG showed modifications in comparison with basal EEG, whereas only 50% of the presyncope positive patients showed slight alterations. There was no EEG alteration for tilt negative patients. The EEG result was markedly different in patients with tilt-induced type 2A syncope in comparison with those with type 1. Type 2A showed the following: 1) slowdown and reduced amplitude of electrical activity during the prodromes; 2) during the syncope, first pseudorhythmic then polymorphic delta activity were followed by total disappearance of activity ("flat" EEG); 3) then, in inverse sequence, reappearance of polymorphic then pseudorhythmic delta activity (average duration of syncope 37 s); 4) lastly, slowdown and reduced amplitude of electrical activity similar to that preceding the syncope. Whereas type 1 revealed: 1) no alteration of electrical activity during the prodromes; 2) during the syncope, first theta then polymorphic delta activity (average duration of syncope 16 s); 3) subsequent normal EEG. CONCLUSIONS: These observations indicate a correlation between the type of tilt test positivity and the EEG results, the latter being markedly more serious in type 2A than in type 1. EEG behavior, different in the two types also during the prodromes and the post-syncopal phase, would suggest a cerebral circle vasoconstriction mechanism in type 2A but not in type 1 mixed with a prevalent vasodepressive component. Should these preliminary results be confirmed by further data there will be evident clinical, prognostic and therapeutic implications. In the light of the considerably different involvement of the central nervous system, we believe it will be necessary to redefine the various types of neurally-mediated syncope in terms of seriousness. Simultaneous EEG could be proposed routinely in tilt test execution and become a determining factor in the choice of a therapeutic option. PMID- 10832127 TI - [The role of the echo-dipyridamole test in the diagnosis of coronary disease in patients with associated aortic stenosis]. AB - BACKGROUND: Coronary vasodilator reserve is often significantly impaired in patients with aortic stenosis by several mechanisms: coronary artery disease, left ventricular hypertrophy, increase in cardiac chamber stiffness. The aim of this study was to evaluate the feasibility and the diagnostic accuracy of the dipyridamole echocardiography test in the diagnosis of coronary artery disease in patients with aortic stenosis. METHODS: Forty patients (26 males, 14 females, mean age 69 +/- 8.9 years) with aortic stenosis (mean valve area 0.7 +/- 0.3 cm2 calculated by the continuity equation) were studied by two-dimensional echocardiography during dipyridamole infusion up to 0.84 mg/kg over 10 min. Wall motion was graded for each segment as normal, hypokinetic, akinetic and dyskinetic. Dipyridamole echocardiography was considered positive for ischemia if wall motion in at least one segment worsened by at least one degree point level compared to wall motion at rest. All patients underwent coronary angiography (mean time after dipyridamole echocardiography 7 +/- 3 days). The chi 2 test and Student's t-test for paired data were used; a p value of < 0.05 was considered as statistically significant. RESULTS: Only one dipyridamole echocardiography was interrupted because of supraventricular tachycardia appearance. Nine patients showed new asynergy areas during dipyridamole echocardiography; 19 patients had ST segment downsloping of > or = 1 mm during dipyridamole infusion; 12 patients experienced angina during the test. Angiography showed a significant coronary stenosis in 10 patients. Dipyridamole echocardiography sensitivity was 80%, specificity was 96%; specificity of ST segment downsloping and angina were 63 and 76% respectively. CONCLUSIONS: Dipyridamole echocardiography in patients with aortic stenosis is safe and feasible with good sensitivity and better specificity. Our study suggests also that dipyridamole echocardiography test is able to rule out patients with aortic stenosis and coronary artery disease as opposed to those with angina without organic stenosis of the coronary vessels. PMID- 10832128 TI - [The usefulness of the association of clinical probability, rapid plasma measurement of D-dimer, compression echography of the lower limbs and echocardiography in the diagnosis of acute pulmonary embolism]. AB - BACKGROUND: The aim of this study was to investigate the diagnostic utility of clinical probability, rapid plasma D-dimer assay, compression ultrasonography (CUS) and transthoracic echocardiography (TTE) in patients with suspected pulmonary embolism. METHODS: One hundred forty consecutive outpatients with suspected pulmonary embolism were enrolled in a prospective trial. We evaluated sensitivity, specificity, positive and negative predictive value of a combination of clinical probability, D-dimer, CUS and TTE using perfusion lung scan and pulmonary angiography as a combined gold standard for determining the presence or absence of pulmonary embolism. Clinical probability was assessed in accordance with the PIOPED criteria. The D-dimer (Nycocard) level was considered as abnormal > 0.3 mg/l, the CUS if incompressibility of the leg veins was showed, and the TTE if right ventricular dilation was present, in the absence of chronic pulmonary disease. The combination of these tests was considered consistent with the presence of pulmonary embolism if D-dimer plus CUS and/or TTE showed abnormal results. A pulmonary embolism was excluded if D-dimer and CUS showed normal findings or a low clinical probability was associated with normal findings of CUS and TTE. RESULTS: One hundred eleven patients were evaluated. Pulmonary embolism was present in 45/111 (40%) patients. The combination of tests showed positive findings in 39/39 patients with pulmonary embolism, negative findings in 47/50 without pulmonary embolism and non-diagnostic results in 22/111 (20%) patients (95% confidence interval--CI 12-28%). There were three false positive and no false negative results. Sensitivity and specificity were 100 and 94% respectively (95% CI 92-100% and 87-100%); positive and negative predictive values were 93 and 100% (95% CI 85-100% and 93-100%). None of these tests, separately, showed enough sensitivity and specificity. CONCLUSIONS: The combination of clinical probability, D-dimer, CUS and TTE was highly accurate to confirm or rule out pulmonary embolism. PMID- 10832129 TI - [A cardiac embolic stroke due to malposition of the pacemaker lead in the left ventricle. A case report]. AB - Misplacement of a permanent pacemaker lead has been described in several locations but rarely in the left ventricle. Less commonly, as described in our report, malposition may occur when the lead perforates the interatrial septum and extends across the left atrium and through the mitral valve into the left ventricle. The actual incidence of this pacemaker complication is unknown. Consequences may include perforation and systemic or cerebral thromboembolic events. We report the case of a patient with unintentionally misplaced left heart pacemaker lead admitted for neurological symptoms consistent with embolic stroke. The patient was on aspirin when symptoms occurred. The lead misplacement was not recognized at the time of implantation. Pacing threshold was normal. A 12-lead electrocardiogram showed right bundle branch block configuration paced complexes. By two-dimensional and transesophageal echocardiography, the pacemaker lead was carefully evaluated. The pacemaker lead crossed the interatrial septum, the left atrium, the mitral valve to be implanted in the left ventricular endocardium. To avoid the risk of future embolization, it was felt that the lead should be removed and right ventricular pacing established, once anticoagulation treatment was initiated. Successful percutaneous lead replacement was accomplished without sequelae. Measures to avoid lead misplacement are suggested. PMID- 10832130 TI - [The correction of pulmonary insufficiency with a cryopreserved homograft: an optimal solution to a postoperative problem not rare]. AB - Although the results of surgical repair for congenital pulmonary stenosis are generally good, some patients develop progressive symptoms related to pulmonary regurgitation and right ventricular dilation. Pulmonary homograft implantation may have a beneficial effect on these symptoms, due to a reduction in the volume overload of the right ventricle and hemodynamic improvement. We describe our experience of one patient with severe pulmonary regurgitation following pulmonary valvotomy performed with the Brock technique during childhood because of pulmonary valve stenosis. The patient was admitted to our Institution because of dyspnea on exertion (NYHA functional class II-III) and paroxysmal episodes of supraventricular arrhythmias. Echocardiography showed severe pulmonary regurgitation, an important right ventricular dilation associated with severe tricuspid insufficiency and a patent foramen ovale without any significant shunts. Surgical repair was performed through a median sternotomy with cardiopulmonary bypass and moderate hypothermia. The right ventricular infundibulum was opened and a cryopreserved pulmonary homograft was implanted with continuous sutures. De Vega annuloplasty was performed on the tricuspid valve and the patent foramen ovale was closed with a running suture. Postoperative course was uneventful and the patient was discharged on the seventh postoperative day. Three months after surgery the patient is asymptomatic and echocardiographic evaluation shows no evidence of pulmonary or tricuspid regurgitation, a decrease in right ventricular dilation and a significant improvement in biventricular systolic and diastolic function. In conclusion, pulmonary regurgitation after surgical valvotomy can be treated with the implantation of a cryopreserved pulmonary homograft with satisfactory results. It would appear advisable to perform surgical repair of concomitant right heart anomalies, such as secondary tricuspid insufficiency, to obtain both a decrease in right ventricular overload and a regression of its preoperative dilation. PMID- 10832131 TI - [An oligosymptomatic giant left atrial myxoma]. PMID- 10832132 TI - [Has the time come that the cardiologist may prescribe "a glass of wine daily, better if red" in the prevention of cardiovascular diseases?]. PMID- 10832133 TI - [Heart failure due to diastolic dysfunction: the treatment principles]. AB - The clinical relevance of diastolic dysfunction in heart failure has recently been emphasized. In fact, the presence of signs of heart failure does not imply a depressed left ventricular systolic function; moreover, the severity of heart failure and effort tolerance are more closely related to diastolic than to systolic indexes. However, the principal trials about the treatment of heart failure were mainly addressed to patients with significant left ventricular systolic dysfunction, whereas the optimal therapy for diastolic dysfunction is not well known. The aim of this review was to assess the rationale and the therapeutic options in heart failure due to diastolic dysfunction. A diastolic dysfunction can be exclusive or associated with systolic dysfunction, as in dilated cardiomyopathy. It has to be noted that in this disease an improvement of diastolic function was demonstrated for most of the drugs currently employed in the treatment of heart failure, such as vasodilators, ACE inhibitors, beta blockers, digitalis, and other inotropic drugs. Moreover, the favorable effect of the treatment on diastolic parameters (reduction of left ventricular filling pressure, regression of restrictive filling pattern) is associated with a positive prognostic impact. The main objective of the treatment of heart failure with preserved left ventricular systolic function is to control the symptoms by means of lowering high left ventricular filling pressure without significantly lowering cardiac output. According to the therapeutic guidelines of the American College of Cardiology/American Heart Association Task Force, the drugs indicated to treat symptomatic patients with heart failure and preserved left ventricular systolic function are diuretics and nitrates. Potentially useful, but with insufficiently proven efficacy are beta-blockers, calcium antagonists and ACE inhibitors, whereas direct vasodilators and inotropic drugs were considered inadvisable. It is important to remember that the treatment might possibly be oriented to the cause and also to the possible precipitating factors of the heart failure syndrome (i.e. ischemia, tachycardia, arrhythmias, hypertension). In conclusion, considering the relatively common incidence of heart failure due to prevalent diastolic dysfunction, and the few available data about the therapeutic options in these patients, large multicenter trials devoted to the treatment of this syndrome are needed. PMID- 10832134 TI - [The diagnosis of ischemic cardiopathy in women]. AB - Risk stratification and therefore evaluation of pre-test probability of coronary artery disease represent an important point in order to choose the ideal noninvasive test in women. Risk evaluation may be performed in relation to the presence of so-called major (hypertension, diabetes, age > 65 years) or minor (dyslipidemia, smoking abuse) risk factors. The standard ECG exercise test represents an important step, but only if easily interpretable and feasible: a complete negative test suggest the absence of coronary artery disease; its positivity should indicate the opportunity to perform coronary angiography in the presence of moderate to severe pre-test probability. If ECG exercise test is not feasible, we would suggest a test of imaging; in the case of high pre-test probability we would recommend a test with high specificity, such as echo-stress test, whereas in the case of lower pre-test probability we would prefer a test with high sensitivity, such as myocardial scintigraphy. PMID- 10832135 TI - [The heart patient in the mountains: the behavioral indications]. AB - Even today, a restrictive attitude frequently leads the physician to discourage the patient with cardiovascular abnormalities from sojourning at an altitude over 1000 m. This attitude, however, is not supported by any scientific proof. After a review of the major cardiovascular adaptations during high altitude exposure, we reported the principal studies of the effects of high altitude on patients with coronary heart disease, particularly during exercise. On the basis of personal experience and of other authors, we can state that the patient with ischemic heart disease, asymptomatic and with a recent clinical and functional evaluation, can stay in the mountains, even at altitudes of 2000-3000 m, and can hike and ski (cross-country and downhill). The risks appear to be related to factors independent of altitude, such as excessive cold or intense emotional stress due to dangerous situations, conditions that must consequently be avoided. On the other hand, an individual approach should be followed for the other congenital and acquired heart diseases, in which case we must consider, in addition to the clinical situation, some peculiar aspects of mountain environment (hypoxia, isolation, difficult access to medical facilities). Finally, the hypertensive patient, expected to have higher blood pressure values, especially diastolic, should frequently record his values during the first week at high altitude, eventually adjusting the therapy. PMID- 10832136 TI - [A resources-activities analysis in cardiology: a report on Piemonte experience. The Cardiology Network of the Piemonte Section of the Associazione Nazionale Medici Cardiologi Ospedalieri]. AB - We are living in an era characterized by cost-containment. Thus, expenditure for the health system represents one of the most important items of the national balance. Nowadays, the optimal management of health organization is essential. In addition, patients expect hospitals to provide more and more effective and efficient cures. With respect to this need, in order to achieve high standard of health cure, the identification of work and the time that the medical departments dedicate to a specific therapeutic and/or diagnostic procedure represent a fundamental tool for the optimal management of patients. However, to provide reliable results, these data should be collected in the largest number of structures possible. For this purpose, 17 departments of cardiology have organized a network with the goal of creating the most important data base on this matter available in Italy. The evaluation of the data obtained (more than 8000 medical procedures) allows us to identify the resources normally used for curing patients admitted to cardiological departments. These provide an extensive view of the work load in the Public Health Services fundamental to implementing a health service in accordance with the criteria of Total Quality Management. The present paper illustrates the scope of this study and the methods used to create the data base. In addition, it discusses some cultural aspects related to the introduction of managerial criteria in the medical arena. The detailed results, regarding the items analyzed, will be presented in a special paper. PMID- 10832137 TI - [The echo-stress test with dipyridamole-atropine on the 3rd-5th day of an uncomplicated acute myocardial infarct for risk stratification and early discharge]. AB - BACKGROUND: The aim of this study was to evaluate if dipyridamole-atropine stress echocardiography (DASE) performed between the third-fifth day in uncomplicated acute myocardial infarction allows for an effective risk stratification with an early discharge in some cases. METHODS: Between February 1997 and September 1998, 190 patients (138 males and 52 females, mean age 59 +/- 10.3 years), with acute myocardial infarction, were enrolled in the study. DASE was performed between the third-fifth day with a dipyridamole infusion of 0.84 mg/kg over 10 min followed by 1 mg of atropine from the twelfth to the fifteenth minute. DASE was considered positive in the presence of a new or worsening dyssynergy. Patients with heart failure, angina, major arrhythmias, and poor acoustic window were excluded. In the follow-up spontaneous events were defined as cardiac death, non-fatal myocardial reinfarction, unstable angina or heart failure (with hospitalization). RESULTS: DASE was performed in 92 patients (48.4%), all without complications: 29 patients (31.5%) had a negative DASE result, and 63 patients (68.5%) had a positive DASE. The average hospital stay of patients with a negative test was significantly lower in comparison with that of patients with a positive test (7.55 +/- 1.32 vs 9.29 +/- 1.61 days, p < 0.0001). Events occurred in 19 patients (20.6%), 2/29 patients with a negative DASE (6.9%), 17/63 patients with a positive DASE (27%), 6/43 patients with homozonal positivity after atropine or high-dose dipyridamole (14%), 11/20 patients with heterozonal positivity or homozonal positivity after low-dose dipyridamole (55%). On univariate analysis the variables significantly associated with spontaneous events were: age (chi 2 = 6.41, p = 0.019), left ventricular ejection fraction at rest (chi 2 = 8.89, p = 0.004), number of asynergic segments after stress (chi 2 = 6.87, p = 0.010), increase in the number of asynergic segments after stress (chi 2 = 4.01, p = 0.039), wall motion score index after stress (chi 2 = 9.60, p = 0.003), increase in wall motion score index after stress (chi 2 = 3.60, p = 0.049), DASE positivity (chi 2 = 4.89, p = 0.029), homozonal positivity after low-dose dipyridamole (chi 2 = 8.57, p = 0.013), heterozonal positivity (chi 2 = 13.10, p = 0.001). On Cox's multivariate analysis independent predictors of events were: age (relative risk 3.92, p = 0.0146), DASE positivity (relative risk 1.79, p = 0.0054). CONCLUSIONS: DASE between the third-fifth day in uncomplicated acute myocardial infarction is feasible, tolerable, safe, and effective for early risk stratification. A negative DASE detects a "very low-risk" patient group, and allows for an earlier hospital discharge, without an increased risk of events. The heterozonal positivity or the homozonal positivity after low-dose dipyridamole indicates the need for a coronarography, due to the high risk of events at follow-up. PMID- 10832138 TI - [The cardiologist facing pulmonary embolism. The experience of 160 cases of acute cor pulmonale]. AB - BACKGROUND: The results of recent multicenter studies dealing with pulmonary embolism often reveal remarkable discrepancies in terms of diagnosis, prognosis and treatment, partly due to the heterogeneity of study patients and of evaluation criteria. Our prospective study focused exclusively on patients affected by pulmonary embolism with a hemodynamic pattern of acute cor pulmonale, investigated at a single center. Particular attention was paid to in-hospital mortality, embolic recurrences, major bleeding and underlying pathologies. METHODS: This study includes 160 cases (103 women with a median age of 71 years and 57 men with a median age of 65 years) in whom the clinical and echocardiographic findings suggestive of acute pulmonary embolism were confirmed by lung perfusion scan, pulmonary angiography, techniques for the detection of deep vein thrombosis and/or autopsy. RESULTS: The most common clinical manifestations were: dyspnea (92% of cases), tachycardia (80%), syncope (44%), cardiac arrest (22%), and shock (20%). Thoracic pain was present in only 27% of patients. None of the patients showed a normal ECG; a right bundle branch block was found in 47% of cases. T-wave inversion in the precordial leads (32%) was not related to the severity and outcome of pulmonary embolism. Present or previous deep vein thrombosis was found in 53 and 26% of cases, respectively. Only in 2 patients pulmonary embolism was secondary to a deep vein thrombosis of the upper limbs. Intravenous heparin alone was used in 36% of cases, whereas 56% were treated with thrombolytic agents + heparin. Major bleeding occurred in 9% of patients treated with heparin alone, and in 16% of those who received heparin + thrombolytic drugs. Death occurred in 17% of the former, and in 27% of the latter patients. The in-hospital mortality rate was related not only to the presence of cardiac arrest and--to a lower degree--of shock, but also to the recurrence of pulmonary embolism and to the underlying heart disease. No relationship was found between mortality and age, intracardiac thrombi or malignancy. Prognosis was quite different depending on clinical presentation, with a death rate ranging from 11% in the absence of systemic hypertension, and 77% in the presence of cardiac arrest. CONCLUSIONS: Even the "massive" pulmonary embolism that is observed in a cardiac department represents a true "spectrum" of pathological conditions, a spectrum that should be taken into account not only in order to evaluate prognosis and treatment in a particular case, but also when meta analyses are performed. PMID- 10832139 TI - [Reconstruction of the aortic root: midterm results]. AB - BACKGROUND: The aim of this study was to assess mid-term results of aortic reconstruction. METHODS: According to Dr. David's technique, 20 patients (17 males, 3 females, median age 63 years) were examined, who underwent surgery between September 1996 and August 1999. Indications for operation were aortic insufficiency with ascending aorta aneurysm in 19 patients, and acute Stanford type A dissection in 1 patient. In every patient the native valve was preserved and suspended inside a tubular prosthesis on which the coronaries were reimplanted. RESULTS: No patient died in hospital. Follow-up (mean 13 months) was complete for all patients. One patient died of extracardiac causes. Seventeen of 19 survivors are in NYHA functional class I, the remaining 2 in class II. Nineteen patients underwent echocardiography which showed moderate aortic regurgitation in 1, mild in 6 and absent or trivial in 12. There were no instances of thromboembolism. CONCLUSIONS: These results are encouraging and have brought us to consider aortic reconstruction a valid alternative to traditional aortic root replacement. Dr. David's technique is reproducible and reliable in the long term. PMID- 10832140 TI - [Reconstructive surgery of the mitral and tricuspid valves with a Cosgrove Edwards flexible ring]. AB - BACKGROUND: Mitral and tricuspid valve asymmetric annular dilation represents the most important mechanism which produces insufficiency. Recent computerized in vitro and in vivo three-dimensional models have been developed in order to better understand the competing factors (annular dilation, displacement of papillary muscles, left and right ventricular geometry). The leading cause of mitral and tricuspid competence is a sphincteric action of both annuli, during systole and diastole, the loss of which produces asymmetric dilation and therefore the absence of cusp coaptation. The Cosgrove-Edwards dynamic ring corrects, alone or in combination with other procedures on the valves, this patho-anatomic feature in a physiological way by restoring the normal annular dimensions and the sphincteric movements during the cardiac cycle. METHODS: Between June 1998 and May 1999, 30 adult patients underwent mitral (n = 20, Group I) or tricuspid valve repair (n = 10, Group II). Regurgitation was due to a degenerative disease in 13 Group I patients and to ischemic (n = 3), congenital (n = 2) or dilated cardiomyopathy (n = 2) in the others. In Group II the leading cause of insufficiency was functional regurgitation in 7 patients and organic in 3. Associated procedures were carried out in 4 Group I patients and in all Group II patients. Regurgitation was evaluated by transesophageal echocardiography before, during and 3 months after operation. The maximal regurgitant area (MRA) and the grade of insufficiency were evaluated using the equation: MRA < 2 cm2 = grade 0, MRA > 2 < 4 cm2 = 1+, MRA > 4 < 7 cm2 = grade 2+, MRA > 7 < 10 cm2 = 3+, MRA > 10 cm2 = 4+. RESULTS: The operative mortality was 0%. One Group I patient died 3 months after operation due to bronchopneumonia. No patient was reoperated on for plasty failure in both groups during the follow-up. Mitral insufficiency was absent (grade 0) in 17 Group I patients and mild (grade 1+) in 3 at the end of operation. At 3-month postoperative transesophageal echocardiographic control mitral insufficiency was absent in 14 patients, mild (1+) in 4 and moderate (2+) in 2. MRA was 3 cm2 in the 2 patients operated on for dilated cardiomyopathy and < 3 cm2 in the others. Preoperative tricuspid insufficiency of grade 4+ in all Group II patients became absent in 9 of them either at the end of operation or at 3-month postoperative control. CONCLUSIONS: The Cosgrove-Edwards dynamic ring as isolated device or in combination with other plasty mitral or tricuspid procedures is a safe, simple, and reproducible method to restore the distorted motion of valvular annuli. It preserves the sphincteric mechanism of the valve and allows for the coaptation of cusps. Although in a small number of patients and for a short period of follow-up our experience corroborates what other more consistent series of patients operated on have shown. PMID- 10832141 TI - [Coronary heart surgery in women: the risk factors and short-term results]. AB - BACKGROUND: Coronary artery bypass grafting is reported to have a higher (2-3 times) mortality in women than in men, most likely due to older age, higher incidence of hypertension, diabetes mellitus, hypercholesterolemia, obesity, angina and preoperative myocardial infarction, smaller diameter of coronary arteries, and smaller body surface area. METHODS: From January 1992 to December 1997, 347 female and 2098 male patients were submitted to isolated coronary artery bypass grafting. For both groups were considered: a) clinical presentation (age, height, weight, body surface area, NYHA and CCS functional classes, incidence of preoperative myocardial infarction); b) risk factors for cardiovascular diseases (diabetes mellitus, smoking habit, dyslipidemia, hypertension, familiarity); c) concomitant diseases (obesity, chronic obstructive pulmonary disease, peripheral vascular disease, thyroid dysfunction); d) hemodynamic and anatomical data (extent of coronary artery disease, diameter of coronary arteries, left ventricular function); e) surgical procedure (number and type of grafts used, urgent procedures, incidence of redo procedures). Early (up to 30 days after surgery) results were evaluated in terms of complications and mortality. RESULTS: On admission, women were older than men (p = 0.0001), were shorter (p < 0.0001), weighed less (p < 0.0001), and had a smaller body surface area (p < 0.0001); they had more severe angina (p = 0.002), diabetes mellitus (p = 0.002), hypercholesterolemia (p = 0.003), thyroid dysfunction (p < 0.0001), their coronary arteries were smaller (left anterior descending artery, p = 0.05; obtuse marginal branch, p = 0.008; diagonal branch, p = 0.01), and had less grafts implanted at surgery (p = 0.02). There was no difference between women and men in the use of the internal thoracic artery. Women did not have a higher mortality than men (4.6 vs 3.2%). Uni- and multivariate analysis did not show extraoperative risk factors for women; for men older age (p = 0.005) and poor left ventricular function (p = 0.01) were independent predictive factors of operative mortality. CONCLUSIONS: In spite of what is suggested by the literature, coronary artery bypass grafting does not have a significant higher operative risk for women than men, probably due to surgical technique refinements and extensive use of the internal thoracic artery. PMID- 10832142 TI - [The ethics of appropriateness and the appropriateness of ethics. Comments on a current topic of which we know little and which we discuss even less]. AB - Doctors and patients "alone on a desert island or in health market?". Antinomy is strongly paradoxical and provocative to invite us to think on two extreme lines of thinking modalities about fragile, variable, and changed relationship between doctors and patients in the light of the new health business administration. Probably, the balance point exists and may be found only introducing three main elements into the complex problem: patient in the middle, health care accuracy, ethical behavior, not only of the single doctor but also of the overall health system. PMID- 10832143 TI - [Multiple rhabdomyomas in a newborn infant. A clinical case report]. AB - Rhabdomyoma is a rare primary benign cardiac tumor usually diagnosed in newborn and infancy. The authors report a case of multiple and completely asymptomatic rhabdomyoma, diagnosed by echocardiography. PMID- 10832144 TI - [The guidelines of the ANMCO-SIC-AIAC on the appropriate use of the methods of permanent cardiac electric stimulation. Associazione Nazionale Medici Cardiologi Ospedalieri-Societa Italiana di Cardiologia-Associazione Italiana di Aritmologia e Cardiostimolazione]. PMID- 10832145 TI - [Light to moderate alcohol consumption and the risk of stroke among American male physicians]. PMID- 10832146 TI - [Familial history of ischemic cardiopathy is associated with endothelial dysfunction of the coronary area]. PMID- 10832147 TI - [The prognostic value of an ergometric scoring system in symptomatic patients with nonspecific ST-T anomalies in the resting ECG]. PMID- 10832148 TI - [The treatment of acute coronary syndromes of the "non-ST elevation" type with enoxaparin: the TIMI 11B study. Thrombolysis in Myocardial Infarct]. PMID- 10832149 TI - [The benefits of the implantable cardioverter-defibrillator in comparisons with anti-arrhythmia agents in patients surviving malignant ventricular tachyarrhythmias: a subanalysis of the AVID study by varying degrees of left ventricular dysfunction. Antiarrhythmic versus Implantable Defibrillators]. PMID- 10832150 TI - [Out-of- or in-hospital occurrences of malignant ventricular tachyarrhythmias: the prognostic significance in the population enrolled in the AVID study. Antiarrhythmic versus Implantable Defibrillators]. PMID- 10832151 TI - [False asystole after the use of manual defibrillators. Do not use heart rate monitoring via the gel pads after the first shock]. PMID- 10832152 TI - [The history of the SET-UP study (or how difficult is neutrality in clinical research)]. PMID- 10832153 TI - Psychiatric emergency services. Introduction. PMID- 10832154 TI - The psychiatric emergency service: where we've been and where we're going. AB - The Psychiatric Emergency Service (PES) has evolved into a separate service with its own space and staff specialized for the handling of psychiatric emergencies. A study of trends in our PES reveals increased need for children's services, issues with managed care and an expansion in the use of the PES as a filter for the mental health system in dealing with substance abuse. Education and research have been added to the missions of the PES and there is strong potential for future development in this area. PESs of the future may be very different, with advances in communication, safety, computerized records and databases. New dilemmas in balancing the patient's right to confidentiality and autonomy against the potential of these advances are bound to occur. PMID- 10832155 TI - Profile of a comprehensive psychiatric emergency program in a New York City municipal hospital. AB - This paper profiles the services provided, and the patient population treated, in a busy inner city Comprehensive Psychiatric Emergency Program (CPEP) located in Elmhurst, Queens, New York City. For each CPEP component, including the emergency room, extended observation unit and crisis services two years of data are reviewed. A diagnostic profile of patients seen, description of services, patient referrals and dispositions are presented. The children and adolescents treated in the CPEP are described in more detail, focusing on the high frequency of violence to self or others seen in their presenting problems. The CPEP's role in providing comprehensive community based services is discussed. PMID- 10832156 TI - The New York Presbyterian Pediatric Crisis Service. AB - This paper outlines the structure of the Pediatric Psychiatry Crisis Service at New York Presbyterian Hospital, a service that provides twenty-four hour emergency psychiatric evaluation and intervention to children, adolescents and families in northern Manhattan. Structure and staffing of the service, usage of the service and the presentation of three cases addressing high, moderate and low risk crisis patients are discussed. Finally, future challenges facing the Crisis Service are addressed. PMID- 10832157 TI - Perceived psychopathology in a painter's work. AB - R. A. Blakelock was a highly acclaimed 19th Century visionary American painter who spent 25 years in mental hospitals. Students rated random samples of Blakelock's work completed before, during, and after his breakdown. Paintings completed after the start of his long-term hospitalization were rated lower in skill and higher in psychopathology than those done before his breakdown. The difference in perceived psychopathology persists when ratings for skill level are controlled. There were also differences due to training level of raters and whether or not the paintings were presented in color. PMID- 10832158 TI - The Assaulted Staff Action Program (ASAP) and declines in rates of assault: mixed replicated findings. AB - The Assaulted Staff Action Program (ASAP) is a crisis intervention program that has been associated with providing both needed support for employee victims of patient assault and declines in rates of assaults in traditional state hospital and community mental health settings. This study examined the possible role of ASAP and declines in rates of assault in three community-based services impacted by privatization and managed care approaches: community residences, an acute care rural community mental health center, and an urban intermediate/extended care facility. ASAP was associated with a statistically significant decline in the rate of assault in the intermediate/extended care facility. The community residence program was not a true test because of structural issues associated with managed care. The community mental health center appeared to be a true failed replication. The extended care facility's decline in the rate of assault was interrupted by the aftermath of one serious clinical incident during one month and then continued to decline. The implications are discussed. PMID- 10832160 TI - [The ICD-10 and the Unified Diagnoses Code of the Pension Insurance. Guidelines for interpretation of the diagnostic codes within the legislative framework for pension insurance]. AB - The "International Statistical Classification of Diseases and Related Health Problems", Tenth Revision (ICD-10), has to be used by all social security funds in Germany since Jan. 1, 2000. For purposes of the official statistical classification of causes of death, it has been used since 1998. There are a number of fundamental differences between the ICD-10 and the ninth revision. In particular, a considerably higher degree of specification and differentiation warrant a thorough look at the ICD-10 in numerous respects. The Unified diagnoses code of the statutory pension insurance is an extract from the ICD-10 referring specifically to diseases occurring in medical rehabilitation and early retirement contexts. Several new four- and five-digit codes were added for statutory pension insurance purposes, hence are not contained in the original ICD-10. This extract will only turn out a practical tool if the physician is well-qualified in his particular field and is familiar with the sometimes very complex differentiations within the systematical register in volume I of the ICD-10. The article provides an overview of the structure of the ICD-10; deals with major novelties and alterations as well as practical problems; uses various exemplary diagnoses to illustrate a systematical approach in possibly problematical encoding; and recalls several general rules for diagnoses encoding in social medicine. PMID- 10832159 TI - Recent HIV testing among general hospital inpatients with schizophrenia: findings from four New York City sites. AB - BACKGROUND: While widely acknowledged to be an important clinical and public health issue, HIV assessment, counseling, and testing for the seriously mentally ill has not been well studied. OBJECTIVE: To determine what proportion and which inpatients with schizophrenia have been recently tested for HIV. METHOD: A sample of 300 inpatients with schizophrenia were recruited from four general hospitals in New York City over a one year period. After confirmation of diagnosis with a structured interview, and elicitation of sociodemographic and drug use information, medical record review identified recent HIV testing. Bivariate and multivariate analyses were used to identify subgroups more likely to be tested. FINDINGS: Recent HIV testing had been performed for 17% of the sample and was concentrated among those with higher documented risks. The majority of patients remain untested even in groups with direct risks, such as injection drug use, and indirect risks, such as frequent cocaine use in last year. Some evidence was found that white patients at risk may be less likely to be tested than Hispanic or African American patients. CONCLUSIONS: Aggressive efforts are needed to improve knowledge of HIV status among acutely ill patients with schizophrenia. PMID- 10832161 TI - [Considerations for standardization of assessment of performance capacity in rehabilitation medicine]. AB - Specific difficulties in the assessment of reduced performance capacity in patients with gastroenterologic illness led to the development of a standardized and unified assessment method. The following five steps--exemplified for chronic liver-disease--were designed: (1) determination of disease features (and their graduations), leading to reduced performance capacity; (2) construction of a detailed performance capacity model using more concrete factors (e.g. motivation, cognition, cardiovascular-pulmonary system, etc.); (3) deduction of rules between disease features and the concrete factors of performance capacity, describing the expected (unfavourable) limitations; (4) consideration of individual limitations on the level of the concrete factors, and (5) conversion into a computer-aided expert system. The benefit of the method developed lies in prompt and practicable assistance by the expert system and in transparency of the decision processes. Therefore, performance capacity assessment will certainly gain in reliability and objectivity. PMID- 10832162 TI - [Integrated rehabilitation of patients with hand injuries--a model of cooperation between acute and rehabilitation clinics]. AB - Close cooperation between acute and rehabilitation clinics are an exception even today. Although operative reconstruction may have been successful, hand injuries will have poor functional results if rehabilitation is insufficient. With the establishment of multi-disciplinary intensive rehabilitation of patients with complex hand injuries under inpatient conditions, a close cooperation of the Clinic of Plastic and Hand Surgery, Klinikum Wuppertal, and the Orthopaedic Clinic, Rhein-Sieg-Klinik Numbrecht, was created. A good functional outcome of the injured hands with a low percentage of remaining invalidity and an early return to work of the affected patients are the dominant aims of this rehabilitation model. When the initial and reconstructive treatment is finished in the Clinic of Plastic and Hand Surgery, inpatient rehabilitation by an experienced rehabilitation team starts immediately. Clinical control of all patients with complex hand injuries is performed in a weekly consultation hour by the hand surgeons and the rehabilitation team. Weekly reports guarantee close and timely documentation of the clinical course. After the end of inpatient rehabilitation, patients return to work, take part in vocational rehabilitation or, if necessary, continue with ambulant treatment organized and controlled by the acute clinic. PMID- 10832163 TI - [Initiation of rehabilitation after surgery for herniated lumbar disk: pilot study of efficacy from the viewpoint of the surgical hospital]. AB - The initiation of rehabilitation after lumbar disc surgery in Germany is up to the operating hospital. A decision between outpatient and inpatient rehabilitation is possible. Until now this decision is not based on scientific data but on the individual experience of the hospital. This prospective study compares the effect of outpatient and inpatient therapy: In 65 patients medical and psychosocial data were collected shortly before operation, 1 week and 6 months after surgery. Of these patients, 42 underwent inpatient, 21 outpatient rehabilitation, 2 patients renounced rehabilitation. The outpatient and inpatient rehabilitation groups showed no significant difference in testing before or shortly after the operation concerning subjective state of health and somatic findings. Some 6 months later, long term outcome was identical, i.e. the groups did not differ in psychosocial or medical respects. However, the inpatient rehabilitation group had taken significantly longer to return to work. This pilot study stresses the importance of further research on rehabilitation after lumbar disc surgery. It should be shared by surgeons and rehabilitation experts. PMID- 10832164 TI - [Whole-body cryotherapy in rehabilitation of patients with rheumatoid diseases- pilot study]. AB - Cryotherapy as a whole-body cold therapy (with cold air cooled by addition of nitrogen blown on the patients in an open cabin) for treatment of inflammatory rheumatic diseases already started in Bad Sackingen in 1986. In 1996, a new cold chamber (this time a closed chamber without any addition of nitrogen) based on compressor technology was introduced. The aim of our study was to test whether significant pain relief could be achieved by means of this cold therapy. Furthermore, we were interested in the practicability and acceptance of this new technique. Wellbeing during the treatment application and pain level were assessed using verbal and numerical rating scales. The sample consisted of 120 consecutive patients (75% women, age: 30-67 yrs, M = 52.6 yrs). These patients were suffering from primary fibromyalgia (40.7%), rheumatoid arthritis (17.3%), chronic low back pain (16.4%), ankylosing spondylitis (10.9%), osteoarthritis (9.1%), secondary fibromyalgia (3.6%) and other autoimmune diseases (1.8%) (mean duration of symptoms: 4 yrs). The patients were treated 2.5 minutes on average in the main chamber (mean temperature: -105 degrees C). The patients' statements concerning their pain level were analyzed by means of analyses of variance with repeated measures and paired-sample t-tests. RESULTS: The pain level after application of the cold therapy decreases significantly. The pain reduction lasts about 90 minutes. The initial pain level decreases during the whole time of treatment, no significant improvement, though, can be shown from the middle to the end of the four-weeks treatment. According to the results of our study, there is evidence that the whole-body cold therapy generates important short-term effects and somewhat weaker effects over the treatment period as a whole. Short term pain reduction facilitates intensive application of physiotherapy and Occupational Therapy. The treatment procedure is practicable, and all in all well tolerated. From the patients' point of view, whole-body cold therapy is an essential part of the rehabilitation programme. PMID- 10832165 TI - [Optimization of the transition from phase II to III of cardiac rehabilitation]. AB - A decisive element of reliable maintenance of the rehabilitation result achieved by patients undergoing inpatient cardiac rehabilitation has turned out to be comprehensive and successful management of the transition into the outpatient phase. This requires concrete planning and implementation of the concept, to be based on patient-related approaches and including the family physician, relatives, occupational institutions, and local and regional partners such as adult education centres, health insurances, sports clubs, psychological individual care, etc. Problem-orientated planning adapted to the individual requirements and expectations of the patient is of utmost importance. PMID- 10832166 TI - [Rehabilitation and the Internet]. AB - An overview is presented of the technical conditions, approaches, existing Internet services, famous homepages about rehabilitation in the Internet (WWW) as well as other possibilities of obtaining information via the Internet. PMID- 10832167 TI - [New developments in patient education. Meeting of the interdisciplinary "Working Group for Patient Education," Rehabilitation Research Promotion Program of the Ministry of Education and Research (BMBF) and German Social Security]. PMID- 10832168 TI - Interdisciplinary collaboration and primary health care reform. PMID- 10832169 TI - Transforming a health care system: managing change for community gain. AB - The research undertaken describes a regional health authority's approach to managing a shift away from acute hospital care towards more community-based, health promoting service orientations in line with new legislated responsibilities. It builds on earlier research of province-wide efforts to create new ways of organizing work within new regional health authority structures. The initial study explored "what matters most", to managing the restructuring of Alberta's health care system and identified ten variables as critical to the transition to regionalization in Alberta. Four were seen to be pre-requisites to effective change: sustaining political will; pacing; resourcing; and, committing to change. Six others were described as continuous process variables; leading; communicating; informing; learning; planning; and, adjusting. This case study looks closely at the operation of these variables in a specific change process within the wider context of the Alberta health reforms. PMID- 10832170 TI - Restructuring public health in Ontario: implications for heart health promotion. AB - Community-based heart health promotion is viewed as an effective means of reducing cardiovascular disease risk. Although public health agencies have a central role in the implementation and dissemination of heart health programmes, their effectiveness is being challenged by major structural changes to Provincial public health systems across Canada, although the impacts of the changes have received relatively little attention in the research literature. As part of the Canadian Heart Health Initiative--Ontario Project (CHHIOP), this study used a qualitative approach to address the perceived implications of these changes to Ontario's public health system for heart health promotion. Interviews (n = 38) were conducted in eight public health units with staff most familiar with managing and/or delivering heart health activities. The results are mixed; that is, while many see the future of heart health promotion programming in Ontario as being at risk, others see recent changes as a step forward toward their institutionalization, particularly in light of recent funding decisions made by the Ministry of Health's Health Promotion Branch. PMID- 10832171 TI - The experiences of women with heart disease: what are their needs? AB - PURPOSE: To identify experiences and needs of women with coronary artery disease. METHODS: Qualitative research utilizing focus groups and grounded theory. Women were recruited from the University of Ottawa Heart Institute Alumni, a group of former Heart Institute patients, and from the Heart Institute cardiac rehabilitation program. FINDINGS: All women had difficulties recognizing their symptoms as heart disease and 35% reported they were initially misdiagnosed. In the post-diagnosis period, the experiences of older and younger women differed. For women over 60, maintaining their functional capacity, memory problems and a focus on resiliency were the main issues. For women under 60, emotionally focussed concerns were predominant. Both groups had to contend with over protective family members. CONCLUSIONS: These findings could contribute to the development of more effective public education campaigns. They also highlight the need for programs and research in the area of women's heart health and rehabilitation that address emotional aspects of the illness. PMID- 10832172 TI - The costs of cardiorespiratory disease episodes in a study of emergency department use. AB - OBJECTIVE: The average per person direct cost of illness of cardiorespiratory disease episodes was estimated based on a prospective study of emergency department visits. METHODS: Economic modelling of health care costs using prospectively collected resource utilization data (9/1/94 to 8/31/95) from hospital emergency department visitors assigned a diagnosis of asthma, chronic obstructive pulmonary disease (COPD), respiratory infections or cardiac conditions. RESULTS: The total direct costs (1997 CDN$) [95% C.I.] per patient were $1,043.55 [$922.65, $1,164.47] for asthma, $1,690.11 [$1,276.92, $2,103.30] for COPD, $676.50 [$574.46, $778.54] for respiratory infections, and $3,318.74 [$2,937.72, $3,699.76] for cardiac conditions. CONCLUSIONS: This study showed that on average, patients diagnosed with a cardiac condition had the highest total direct cost. Hospitalization cost was the largest component of costs for all diagnoses except asthma, for which medications were the single largest component of direct costs. PMID- 10832173 TI - Beyond administrative data: characterizing cardiorespiratory disease episodes among patients visiting the emergency department. AB - We attempted to address deficiencies in administrative health service data during a study of cardiorespiratory emergency department visits. From 1994-1996, we obtained data on 9,264 visits and conducted 1,772 follow-up interviews. The median interval between symptoms and visit ranged from 0.8 days (95% CI 0-1.7) for cardiac conditions to 4.0 days for chronic obstructive pulmonary disease (COPD) (95% CI 2.7-5.3) and respiratory infections (95% CI 3.5-4.5). Infection was the most common trigger of respiratory visits. Although most had improved at follow-up, symptoms persisted following the visit for a mean of 4.5 days (95% CI 3.8-5.4) for cardiac conditions to 8.4 days (95% CI 7.2-9.5) for COPD. Among adults aged < 70, the mean number of reduced activity days per episode ranged from 4.7 (95% CI 3.9-5.4) for asthma to 6.6 (95% CI 5.9-7.4) for respiratory infections. Our data assist in interpreting epidemiological studies based on administrative data, and illustrate the broad impacts of cardiorespiratory disease episodes. PMID- 10832174 TI - Waiting for a diagnosis after an abnormal screening mammogram. SMPBC diagnostic process workgroup. Screening Mammography Program of British Columbia. AB - BACKGROUND: Women with abnormal screening mammograms require diagnostic assessment and experience anxiety until a diagnosis is established. This report evaluated the timeliness of diagnosis after an abnormal screening mammogram in the Screening Mammography Program of British Columbia (SMPBC). METHODS: Information on diagnostic interventions following an abnormal screen (N = 10,314) provided through 11 regional SMPBC services between January 1, 1993 and June 30, 1994 were abstracted and analyzed. RESULTS: The median time from abnormal screen to diagnosis was 3.4 weeks with regional variation of 2.0 to 4.7 weeks; 10% waited 8.7 weeks or longer. For the 19% of women proceeding to open biopsy, the median diagnostic interval was 7.1 weeks with regional variation of 4.6 to 9.3 weeks; 10% waited 13.1 weeks or longer. INTERPRETATION: After an abnormal screening mammogram, women waited many weeks for a definitive diagnosis, especially those proceeding to open biopsy. Opportunities for process improvement were identified. PMID- 10832175 TI - Recent trends in provincial life expectancy. PMID- 10832176 TI - An assessment of health technology assessment in Canada. PMID- 10832177 TI - Land and sea connection: the east coast fishery closure, unemployment and health. AB - The closure of the east coast fishery in Canada in 1992 has affected the lives of individuals and communities both directly and indirectly, causing massive unemployment. Previous research indicates that unemployment negatively affects people's health and other aspects of family functioning. This study assesses the perceived effects of unemployment on the individuals' health, and compares the psychological well-being of the employed and that of the unemployed individuals in the same communities affected by the cod moratorium. The sample consisted of 681 individuals, 16 years of age and older from randomly selected households. They were individually interviewed using structured questionnaires. The GHQ-28 with a high test-retest coefficient (0.90) and split-half reliability (0.92) was used to measure the psychological well-being. The unemployed reported more stress, boredom, high level of uncertainty, less satisfaction with themselves, their life, their educational level, income and health. Implications for health promotion, policy formulation and future research are put forward. PMID- 10832178 TI - Modern antiretroviral therapy improves life expectancy of gay and bisexual males in Vancouver's West End. AB - This study was undertaken to evaluate the life expectancy of gay and bisexual men in the West End of Vancouver, British Columbia during two time periods. Mortality data for males were obtained for the periods 1990 to 1992 and 1995 to 1997 and population estimates were obtained from the 1991 and 1996 Census. The proportion of the male population over 20 years of age estimated to be gay and bisexual was derived from a random telephone survey. Mortality patterns were assessed by comparing changes in life expectancy at age 20 years between the periods, and by examining the life expectancy lost attributed to HIV/AIDS. Between the periods there was 3.8 +/- 3.4 years increase in life expectancy among gay and bisexual men. At exact age 20 years, life expectancy increased from 37.0 +/- 3.5 years during the period 1990 to 1992 to 40.8 +/- 2.4 years during the period 1995 to 1997. The loss of life expectancy attributed to HIV/AIDS at this age was 13.8 +/- 3.9 during the first period and 9.8 +/- 3.6 years during the second period. This gain is most likely the result of the improved efficacy of antiretroviral therapies. PMID- 10832179 TI - HIV prevalence remains low among Calgary's needle exchange program participants. AB - OBJECTIVES: To determine the demographic characteristics, risk behaviours and prevalence of HIV-1 among injection drug users (IDU) attending Calgary's needle exchange program (NEP). METHODS: A survey was conducted from June through September 1998 among IDU attending Calgary's NEP. Demographic and behaviour characteristics were determined by personal interview and saliva was tested for HIV antibody. RESULTS: There were 278 participants providing 272 saliva specimens. Nine were positive for HIV-1 (3.3%, 95% C.I. 1.6-6.4%). Sexual and injecting practices, cities where drugs had been used, incarceration, addiction treatment and demographic characteristics were described, and a subanalysis for women, youth and Aboriginals was carried out. CONCLUSIONS: HIV prevalence remains low among NEP attenders in Calgary, although high-risk behaviours are common. Women, youth and Aboriginals have unique risk behaviour profiles. Many IDU want to participate in addiction treatment, and strategies should be made to provide accessible, appropriate treatment services. PMID- 10832180 TI - Cree infant care practices and sudden infant death syndrome. AB - OBJECTIVE: To identify contemporary Cree infant care practices and any risk factors associated with Sudden Infant Death Syndrome. METHODS: A questionnaire conducted and recorded in the homes of 70 Cree women (83% of sample available) with infants under 12 months of age, living full time on a Cree reserve. Participatory observation was also used with mothers who agreed to demonstrate certain features of Cree infant care. FINDINGS: 70% of mothers initiated breast feeding and 43% nursed from 6 to 15 months. Cree infants are tightly swaddled and placed in a supine position for sleeping. Prenatal care is seldom utilized because of cultural beliefs that planning ahead constitutes bad luck during birth or results in deformed infants. CONCLUSIONS: Existing infant care practices include several that are considered protective to an infant in terms of reducing vulnerability to SIDS. Swaddling infants in very warm houses and smoking during pregnancy could contribute to a higher SIDS incidence. PMID- 10832181 TI - A systematic review of the effectiveness of environmental awareness interventions. AB - PURPOSE: To evaluate and summarize evidence on the effectiveness of interventions available to public health staff regarding the protection of the public from environmental risks. METHOD: This systematic review involved a comprehensive literature search, screening for relevance, quality assessment of relevant studies, data extraction and synthesis. RESULTS: Fourteen of 65 relevant studies were of 'moderate' or 'strong' quality. Intervention types in these 14 studies included: mass campaign, counselling, school curriculum, educational sessions, and distribution of printed materials. Short-term improvements in awareness or knowledge were observed in 13 of the 14 studies. Eight of 13 studies that examined behavioural outcomes observed short-term improvements in self-reported behavioural outcomes. CONCLUSIONS: Positive short-term changes in health protective awareness, knowledge and self-reported behaviour appear to be associated with relatively intensive interventions that use multiple methods and settings, and/or are delivered over multiple sessions. PMID- 10832182 TI - Measuring the impact of influenza on the hospital admission rates of the elderly in Ontario: a five-year admission rate analysis, 1988-1993. AB - OBJECTIVE: A retrospective study was conducted to compare age- and sex-specific rates of hospital admission of the elderly in Ontario for five influenza seasons from 1988-1993 for pneumonia, congestive heart failure and chronic respiratory disease. RESULTS: Significant increases in admissions were found for pneumonia in each influenza season for both sexes. Admissions were significantly increased for congestive heart failure for the oldest males in all years; and for four of five years for the youngest two age groups of males. For females significant increases in admissions occurred in the oldest two age groups in each influenza season; and for three of five seasons for the youngest age group. For chronic respiratory disease, significant increases in admissions were found for each influenza season for all years for the two youngest age groups of males and females. CONCLUSION: The impact of influenza is substantial in terms of morbidity in the elderly. PMID- 10832183 TI - Using healthy life years (HeaLYs) to assess programming needs in a public health unit. AB - In December 1997, public health units in Ontario received revised mandatory program guidelines from the Ministry of Health in advance of the downloading of public health to municipalities. Public health units face difficult decisions in allocating municipal resources to meet the Province's mandated programs. To set priorities for resource allocation, it is critical to assess need across program areas and to use a common unit in doing so. This paper applies the Healthy Life Years (HeaLYs) method in assessing health need related to the mandatory programs for the population of Wellington and Dufferin counties in Ontario. The HeaLYs method incorporates duration and severity of ill-health, incidence and mortality in calculating years of healthy life lost (YHLL). For Wellington-Dufferin, the leading causes of YHLL were concentrated in the program areas of chronic disease, injury, and substance abuse and included four areas not addressed in the MPG (suicide, depression, dementia, and osteoarthritis). PMID- 10832184 TI - The perception and communication of risk: a guide for the local health agency. AB - A local public health agency is expected to respond to a wide spectrum of health concerns, the management of which usually requires effective communication of information, and dialogue with concerned communities. Local health departments have not always found this to be a smooth process. This paper begins by reviewing the public's construction and perception of risk, and examines the changing popular view of the health agency with respect to trust and credibility. The health agency's role and responsibilities in the area of risk communication are addressed in the form of a checklist drawn from a wide range of field-tested sources. PMID- 10832185 TI - In clinical trials, should we be blinded or masked? A proposal for becoming aware of the meaning behind words and concepts. PMID- 10832186 TI - How should a patient with an isolated GGT elevation be evaluated? PMID- 10832187 TI - Screening for renal artery stenosis: which patients? Which test? AB - If a patient has clinical clues suggestive of renovascular hypertension such as persistently high blood pressure despite a multiple-drug regimen, it may be reasonable to screen him or her using captopril renography, duplex ultrasonography, or magnetic resonance angiography. PMID- 10832188 TI - Using viral load, CD4+ levels, and clinical response to guide antiviral therapy for HIV. AB - The decision to start or modify antiviral therapy in patients with human immunodeficiency virus (HIV) infection is not based on any single factor. Although HIV RNA levels are the primary guide to therapy, the CD4+ count and clinical response are also important. PMID- 10832189 TI - Recognizing and managing depression in women throughout the stages of life. AB - Depression is twice as common in women as in men, and women often experience different symptoms, a different course, and a different response to treatment. Furthermore, the menses, oral contraceptive use, parturition, menopause, and old age may cause or exacerbate depression. This paper discusses the diagnosis and management of depression in women throughout the stages of life. PMID- 10832190 TI - 'Rescue' angioplasty for failed thrombolysis. AB - Early restoration of patency of the infarct-related artery is a universally accepted goal in the treatment of acute myocardial infarction. However, even with the most successful fibrinolytic regimen only approximately 50% of patients achieve restoration of normal flow at 90 minutes. "Rescue" angioplasty, the mechanical reopening of an infarct-related artery after unsuccessful fibrinolytic therapy, appears to be beneficial. PMID- 10832191 TI - Infective endocarditis: prevention, diagnosis, treatment, referral. AB - Infective endocarditis is a challenge to the primary care physician, who is not likely to see more than several cases a year. With proper diagnosis and treatment, the overall cure rate is over 80% and major complications such as congestive heart failure can be avoided. In some patients, even in some with acute infection, surgical intervention to restore cardiac function significantly improves the outcome. Guidelines for prophylaxis before various surgical procedures are presented. PMID- 10832192 TI - Rapid assessment of rehabilitation options for functional disability. AB - Although functional disability is common in elderly patients, physicians often overlook it or focus on acute illness, perhaps in part because they are unsure how to efficiently address the problem. A simple, stepwise protocol can be used in either an office or hospital setting to rapidly assess functional disabilities and identify potentially useful interventions. PMID- 10832193 TI - Appropriate use of second-generation antihistamines. AB - Antihistamines, one of the most frequently used groups of medications in the United States, are primarily used in treating allergic rhinitis and urticaria and also anaphylactic reactions, pruritus, and symptoms of anxiety. Second-generation antihistamines are safe and effective for treating allergic conditions. Control of nasal congestion may require additional medication. Further studies will determine if second-generation antihistamines can be used for other medical conditions. PMID- 10832194 TI - Cautious optimism for the future of medicine. PMID- 10832195 TI - Attic cholesteatoma with extension into the tympanum. PMID- 10832196 TI - Prolonged ulcerative laryngitis. PMID- 10832197 TI - Endoscopic view of middle turbinate polyps. PMID- 10832198 TI - Electronystagmography: a patient with peripheral vestibular disorder and debarkment symptoms. PMID- 10832199 TI - Tornwaldt's cyst. PMID- 10832200 TI - Indirect laryngeal surgery in the clinical voice laboratory: the renewal of a lost art. AB - Since the advent of precision instruments and safe techniques for direct laryngoscopic surgery under general anesthesia, indirect laryngeal surgery has become very uncommon. A review of the recent literature finds that few authors advocate indirect surgery under topical anesthesia, and many otolaryngologists dismiss this technique as being either of only historical interest or an idiosyncratic method practiced only by a handful of clinicians. The societal mandate for cost-effective healthcare and the availability of relatively low cost, high-quality endoscopes and video equipment warrant a renewed and broader interest in this type of surgery. In this article, we review a series of 27 indirect surgical procedures performed under topical anesthesia in the clinical voice laboratory. We discuss the indications, outcomes, advantages, and disadvantages of this surgery, and we present a brief analysis of its cost effectiveness. We conclude that indirect laryngeal surgery in the clinical voice laboratory is an effective, safe, efficient, and less costly alternative to some procedures routinely performed under general anesthesia. PMID- 10832201 TI - The importance of quantifying skin reactivity in treating allergic rhinitis with immunotherapy. AB - Therapeutic options for the treatment of allergic rhinitis include environmental modifications to decrease exposure to allergens, pharmacotherapy, and immunotherapy for those patients who do not experience satisfactory relief of their symptoms with medical management. Skin testing is the best established and most sensitive indicator of allergic disease. Several techniques are currently in use to identify pertinent antigens in the treatment of inhalant allergies. We describe the various skin testing techniques that are associated with such inhalant allergies. Quantification of skin reactivity to formulate a successful antigen vial for effective immunotherapy is necessary in the management of allergic disease. PMID- 10832202 TI - A father and son with a nonsevere form of Crouzon's syndrome. AB - Crouzon's syndrome is a hereditary autosomal-dominant disorder. In its classic form, patients experience a premature closure of the cranial sutures, which leads to brachycephaly, proptosis, a small maxilla, and anomalies of the external and middle ear. In this report, we describe the case of a father and son who both had a nonsevere form of this disorder. The two men did not have brachycephaly or proptosis, but they did have ptosis and a mixed-type hearing loss. PMID- 10832203 TI - Spontaneous cerebrospinal fluid leakage and middle ear encephalocele in seven patients. AB - Isolated cases of spontaneous cerebrospinal fluid (CSF) leakage with and without middle ear encephalocele have been reported. These leaks are usually accompanied by episodes of recurrent meningitis, hearing loss, or chronic headache. In this article, we report seven new cases of spontaneous CSF leakage. Six of these patients had conductive hearing loss and serous otitis media, and three had recurrent meningitis. Prior to a definitive diagnosis, six patients had received myringotomy tubes, which produced profuse clear otorrhea. Three patients had positive beta-2 transferrin assays. Computed tomography and magnetic resonance imaging confirmed a defect in the temporal bone tegmen. A combined transmastoid and middle fossa surgical approach with a three-layer closure was used to repair the tegmen defect. All patients had a lumbar drain placed prior to surgery. In addition to describing the seven new cases, we review the history of CSF leakage and discuss diagnostic methods, surgical findings, and our recommendations for management. PMID- 10832204 TI - Intralingual dermoid cysts: a report of two new cases. AB - Dermoid cysts of the oral cavity are rare. When they do occur, the most common site is the floor of the mouth. Intralingual dermoid cysts are even more rare, and until now, there were only 15 such reports in the English-language literature. In this article, we describe two additional cases. Magnetic resonance imaging is extremely helpful in establishing a differential diagnosis. Surgical excision is recommended to correct deglutition and speech problems. Its rarity notwithstanding, dermoid cyst should be considered in the differential diagnosis of tongue masses in the younger population. PMID- 10832205 TI - Initial report of a case of carcinosarcoma of the supraglottis. AB - Minor salivary gland carcinosarcoma is a rare malignancy with an ominous prognosis. This report recounts a single case of supraglottic minor salivary gland carcinosarcoma that was treated aggressively. We discuss the peculiarities of the histology, with special mention of the distinction that must be made between mucosal-origin and salivary-gland-origin carcinosarcoma. Carcinosarcomas are aggressive regardless of their origin, but differentiating their origin is important because the prognosis is worse for those that arise in salivary glands than for those that originate in the surface epithelium. This report adds to the literature a new case of minor salivary-gland-origin carcinosarcoma involving the upper airway. It is the first case we could identify in which the supraglottis was involved. PMID- 10832206 TI - Initial report of a cutaneous T-cell lymphoma appearing on the auricular helix. AB - We report a case of cutaneous T-cell lymphoma in which the first sign of disease was involvement of the superior auricular helix. A review of the literature reveals that T-cell lymphoma often presents cutaneously, but it usually does not involve the ear, and an auricular lesion is rarely the first sign. The uncommon nature of this presentation, in addition to the potential need for multiple biopsies for a tissue diagnosis, can lead to a delay in diagnosis. When evaluating skin lesions in the head and neck, a high index of suspicion for cutaneous T-cell lymphoma is a key factor in its early diagnosis. PMID- 10832207 TI - The role of rhinomanometry, acoustic rhinometry, and mucociliary transport time in the assessment of nasal patency. AB - We conducted a study of 60 patients with different nasal pathologies who complained of nasal obstruction. Our goal was to evaluate the reliability of rhinomanometry, acoustic rhinometry, and the measurement of mucociliary transport time in helping make the diagnosis of nasal pathologies. We also sought to discover whether there is a correlation between the findings of these objective tests and the results of patients' own subjective assessments of nasal obstruction. We found that acoustic rhinometry was more specific and more sensitive than rhinomanometry in diagnosing rhinopathies in patients with structural anomalies. Symptom scores as rated by patients on the visual analog scale frequently did not correlate with objective measures, as patients often overestimated the severity of their obstruction. However, for a few patients, there was a correlation between symptom scores and mucociliary transport times. PMID- 10832208 TI - Explaining ciliary function to patients. PMID- 10832209 TI - Ehrlichiosis ... in Hawaii? An unusual presentation. PMID- 10832210 TI - Encoding specificity and problem-based learning (PBL). PMID- 10832211 TI - The Indian child and the new millennium--hope or hype? AB - The challenges for childcare in the new millennium are outlined. The highly successful polio immunization strategy needs to be extended to other activities. The training programs developed for neonatal resuscitation, intensive care and care in oncology should be extended to other specialties. The journal will provide more fellowships and aid in the development of sub specialties in pediatrics. It will be published monthly and will also go online from January 2000. PMID- 10832212 TI - High frequency ventilation: basic concepts and clinical application. AB - High frequency ventilation (HFV) is a mode of therapy that improves gas exchange with pressure fluctuations much lower than that required for conventional ventilation. HFV is efficacious in the "rescue" management of infants with respiratory failure. Although each HFV system has functional characteristics that are design related, it now appears that when used with similar treatment strategies and within functional limitations, similar outcomes can be achieved. Ideally, the clinician or the operator should be familiar with the basic concepts of different high frequency ventilators to achieve maximal benefits from using these devices in infants with respiratory failure. PMID- 10832213 TI - Socio-demographic and maternal determinants of low birth weight: a multivariate approach. AB - A community based study was conducted in the rural areas of Udupi taluk, Karnataka state of South India to identify the socio-demographic, maternal and obstetric determinants of low birth weight. All singleton live births that occurred in the study area during a one year period (October 1991 to September 1992) were included. A total of 2919 singleton child-mother pairs formed the basis of the analysis. Information about social, demographic and economic conditions of the families; maternal factors such as age, parity, quality of antenatal care and previous obstetric history were collected by interviewing the mothers and family members and verifying the available medical records through the field investigators especially recruited and trained for this purpose. Data was analyzed using multiple logistic regression model. Primis, elderly mothers and mothers who had not received good quality antenatal care were found to be more at risk of having low birth weight babies. Other significant determinants were family custom, socio-economic status and environmental sanitation. PMID- 10832214 TI - National immunization day to assess nutritional status of underfives in Chandigarh. AB - National Immunization Days (NIDs) are an additional opportunity to reach a large number of children and can be used to undertake additional activities beside immunization. This study highlights an attempt to assess nutritional status of under five children by using a NID. Seven thousand four hundred and thirteen underfives were selected randomly from urban, rural and slums areas of Chandigarh. Nutritional status was assessed by using weight for age criteria. Prevalence of protein energy malnutrition (PEM) was found to be about 42% while 22.7%, 14.5%, 4.1% and 0.7% children had grade I, II, III and IV PEM respectively. The prevalence of PEM was significantly higher among females (47.6%), in 1-3 years age group (53.80%), in slum area (67%) and children of labour class (60.5%) (p < 0.001). With increase in family size, the prevalence of malnutrition also significantly increased, and decreased with high literacy rate in parents (p < 0.001). The prevalence of PEM in present study (42%) was comparable to a community based survey (51.6%) among pre-school children of an ICDS block of Chandigarh. National immunization days can be successfully used to assess nutritional status of children. This approach can be used by others for screening common childhood problems, preparing normogram for a region or country, administering Vitamin A and educating mothers. PMID- 10832215 TI - Anemia in children: early iron supplementation. AB - Iron deficiency anemia is the most wide spread micronutrient deficiency disorder in the world, being most prevalent in women and young children. The present study was conducted to find out the prevalence of anemia in children and its contributory factors with reference to early iron supplementation. It was a cross sectional study done on 137 children of age 1-2 years in urban slums of Meerut. Prevalence of anemia in the study group was 59.9%. The socio-economic status, mother's educational status, birth weight, sibling order, and type of weaning food did not show any significant relationship with the prevalence of anemia in these children, whereas, weaning time, nutritional status and early iron supplementation had a positive impact on it. Exclusive breast feeding upto 4 months followed by weaning, adequate nutritional status and early iron supplementation have a definite role in prevention of anemia in children. PMID- 10832216 TI - Immunogenicity of plasma-derived hepatitis B vaccine in preterm infants. AB - The objective was to determine whether plasma-derived hepatitis B vaccine is immunogenic in preterm appropriate for gestation babies when administered at birth and to compare the immunogenicity between 5 micrograms and 10 micrograms doses of the vaccine in these babies. Fifty preterm neonates (31-36 weeks gestation) were randomized to receive 5 micrograms or 10 micrograms doses of plasma-derived hepatitis B vaccine at birth, with subsequent doses 1 and 6 months later. Serum specimens were obtained a month after each dose of the vaccine and were tested for antibody to hepatitis B surface antigen (anti-HBs). Thirty six babies (gestation 31-36 weeks), 18 from each group competed the study. While 89.2% of the babies seroconverted, 82.1% achieved seroprotective titres of anti HBS (> 10 mIU/ml). There was no difference between weight, gestational age, age of administration of vaccine and age of estimation of anti-HBs between 5 micrograms and 10 micrograms groups. The difference in the seroprotective rates were not statistically different between the groups (5 micrograms 78.5%; 10 micrograms--85.7%). Although immune response to plasma derived hepatitis B vaccine in preterm babies is suboptimal when the first dose is administered at birth, the full course achieves adequate seroprotective levels. PMID- 10832217 TI - Neonatal gram-negative bacteremia. AB - A 22 months prospective study of neonatal gram-negative bacteremia was undertaken in a 15 bed NICU to find out the incidence and antibiotic resistance patterns. Clinically suspected 1326 cases of neonatal sepsis were studied during this period. More than 25% of the cases were microbiologically positive for sepsis. Among 230 (67.2%) cases of gram-negative bacteremia, the predominant isolates were Pseudomonas aeruginosa (38.3%), Klebsiella pneumoniae (30.4%), Escherichia coli (15.6%) and Acinetobacter sp. (7.8%). Fifty-nine per cent of the neonates were born in hospital while 41% were from community and referral cases. Lower respiratory tract infection, umbilical sepsis, central intravenous line infection and infection following invasive procedures were the most commonly identified sources of septicemia. Prematurity and low birth weight were the main underlying conditions in 60% of the neonates. Total mortality was 32%. Increased mortality was mainly associated with neutropenia, nosocomial infection and inappropriate antibiotic therapy. Resistance was increasingly noted against many antibiotics. The isolates were predominantly resistant to extended spectrum cephalosporins (25%-75%), piperacillin (68%-78%), and gentamicin (23%-69%). The commonest microorganisms causing gram-negative bacteremia were Pseudomonas aeruginosa followed by Klebsiella pneumoniae. The community-acquired bacteremia was mainly due to E. coli. The proportion of preterm and low birth weight babies was significantly high, and the major contributing factor in total mortality. Sensitivity to different antibiotics conclusively proved that a combination of ampicillin + sulbactam with amikacin or ampicillin + sulbactam with ciprofloxacin is most effective. PMID- 10832218 TI - Therapy and drug resistance in malaria. AB - The aim of this study was to highlight the high degree of clinical resistance of P. falciparum and also of P. vivax to chloroquine and also the importance of early diagnosis and prompt treatment. Ninety cases of smear positive malaria aged between 6 months to 14 years were studied with regards to clinical manifestations, management and outcome. Criteria for drug resistance were absence of clearance of parasitemia and/or persistance of fever after 72 hours of therapy. Chloroquin resistance was noted in 15 (62.5%) cases of falciparum malaria and 15 (51.7%) cases of vivax malaria. The resistant and complicated cases were treated with quinine. Four (6.6%) cases were resistant to quinine and responded to artemether. There was no mortality. Early reporting of cases, frequent sampling malarial parasite, prompt diagnosis of falciparum malaria, early institution of appropriate therapy and awareness of choloquine and/or quinine resistance helps in salvaging lives. PMID- 10832219 TI - Role of steroids in treatment of croup. PMID- 10832220 TI - Designing a research study. AB - There are 10 steps to consider in any research study. These steps can be used to perform a critical appraisal of a published paper, or to design your study in the first place. The use of a simple system such as this can ensure that biases in study design are avoided and that an appropriate study is developed. (1) What is the study hypothesis? (2) What is the study type? (3) What are the outcome measures? Is their measurement biased in any way? (4). What is the study factor (the intervention being offered to the children)? Is there bias in this? (5) Is there a possibility of confounding? (6) What are the reference population, source population and study sample? How have the sampling and selection into the study been performed? (7) Study methods that might threaten the internal validity of the study. (8) Statistical considerations. (9) Are the results clinically and socially significant? (10) Will the conclusions of the study be relevant to the kind of patients you see, or communities you work with? PMID- 10832221 TI - Clinical epidemiological principles in bedside teaching. AB - Medical education emphasises the transmission of large amounts of short-lived medical information. Strategies must be developed to generate in the students, attitudes towards the independent search for and critical appraisal of evidences. Two complementary strategies are particularly well suited to promote these types of attitudes: clinical epidemiology and evidence-based clinical practice. Clinical epidemiology (CE) has been defined as "the science of making predictions about individual patients by counting clinical events in similar patients, using strong methods for studies of groups of patients to ensure that the predictions are accurate. The purpose of CE is to develop and apply methods of clinical observation that will lead to valid conclusions by avoiding being misled by systematic error and chance". On the other hand, evidence based practice has been defined as "an approach to the practice of medicine in which the clinician is aware not only of the evidences which support clinical practice, but the strength and soundness of such evidences". The paper discusses the usefulness of clinical epidemiology and evidence-based health practice for rational decision making at the bedside, and also as tools for clinical teaching. An example involving evidence-based strategies for the management of a patient with bronchiolitis in the emergency room is discussed. PMID- 10832222 TI - Interpretation of diagnostic tests. AB - Clinicians can now base clinical decisions on the results of rigorous studies of the performance of diagnostic tests. In selecting the tests, clinicians should take into account their sensitivity (the proportion of patients with the disease who have a positive test result) and specificity (the proportion of patients without disease who have a negative test results). Sensitivity and specificity are affected by the "spectrum" of patients studied i.e. the severity of disease in those with disease and the clinical characteristics of those without disease. Test results are interpreted by predictive values--the proportion of patients with a positive (negative) test result who have (do not have) the disease. Predictive values depend on both test sensitivity and specificity and disease prevalence. The information content of a test is further increased by taking into account where the cut-off point between normal and abnormal is placed, the degree of abnormality of the result, and the results of the preceding tests (done either in parallel or in series). Describing test performance in terms of likelihood ratios facilitates this process. Readers should be able to critique published studies of diagnostic tests which are still far from perfect. PMID- 10832223 TI - Reading a journal article. AB - Journal reading is an important continuing educational activity in terms of physician preference, frequency of use amount of time devoted to it and impact in changing practice. Clinicians trying to obtain information from journals face several tasks, including the need to assess both the quality and the applicability of the information found in published articles. This article describes the general principles for reading, critically, scientific papers that report clinical research results, which include establishing the kind of question that the authors were trying to answer, the type of study done, if the research design was appropriate for the question, and if it was conducted correctly. According to several available guidelines, once the reader has a clear idea of the study question it is necessary to assess the methods described by the authors in order to evaluate whether the research strategy used is likely to have errors resulting from bias and chance (random) variation. If the paper methodology seems to be adequate, the reader should proceed to determine the results of the article and their potential clinical impact. Finally, the reader must establish the extent to which the results are helpful in caring for his or her own patients, which means assessing the external validity and other related issues. The use of these strategies will improve the efficiency of readers for incorporating research results to their practice. PMID- 10832224 TI - How to read a systematic review. AB - A systematic review is a carefully planned and conducted study designed to answer a specific focused clinical question. A systematic review differs from the frequently encountered narrative review article. Systematic review must be completed using a prespecified methodology. The reader must ask eight important questions when reading a systematic review: (i) did the review address a focused clinical question (ii) is it likely that important, relevant studies are missed (iii) were the inclusion criteria used to select appropriate articles (iv) was the validity of the included studies assessed (v) were the assessments of studies reproducible (vi) were the results similar from study to study (vii) what were the overall results and how precise are they and (viii) will the results help in caring for patients. The article also provides sources for systematic reviews. PMID- 10832225 TI - Granulocyte macrophage-colony stimulating factor (GM-CSF) in neonatal neutropenia. AB - Neutropenia in neonates is often associated with sepsis, prematurity and maternal hypertension with increased risk of mortality. We describe two neonates with neutropenia treated with granulocyte macrophage colony stimulating factor. The total and absolute neutrophil counts showed a marked response and led to a favourable outcome. Human granulocyte macrophage colony stimulating factor may be used as an adjuvant therapy for neonatal neutropenia of different aetiologies. PMID- 10832226 TI - Congenital diaphragmatic hernia "occulta". AB - Congenital diaphragmatic hernia usually presents with cyanotic attacks, dyspnoea and dextrocardia, a typical triad which is almost considered pathognomonic. The case records of 10 patients (out of a total of 20) of congenital diaphragmatic hernia presenting beyond the neonatal period were reviewed retrospectively. Age and symptoms at presentation and signs elicited were paid special attention. All the patients underwent operative repair of hernia under general anaesthesia. Given below is an explanation for such a high incidence of late presentation in our series. This nomenclature (Congenital Diaphragmatic Hernia "Occulta") will help clinicians in remembering this entity if and when such patients present to the clinicians. PMID- 10832227 TI - Post traumatic bile ascites. AB - A 4 year old girl with progressive distension of abdomen and previous history of blunt abdominal trauma was found to have bile ascites due to left hepatic duct (LDH) injury. The management with a brief review of literature is discussed below. PMID- 10832228 TI - Post-encephalitic Kluver-Bucy syndrome. AB - Kluver-Bucy syndrome is a rare amalgamation of neurobehavioural signs and symptoms seen infrequently in humans following insult to bilateral temporal lobes. This report presents a case which along with emerging symptoms of Kluver Bucy syndrome, developed Attention-Deficit/Hyperactivity Disorder (ADHD) consequent to viral encephalitis. The case highlights the presentation and management of this syndrome in a six-year-old girl with primary focus on the implications of Kluver-Bucy symptoms in current clinical practice involving neurobehavioural syndromes in children. PMID- 10832229 TI - Seroepidemiology of hepatitis A in Delhi. AB - A prospective study was done to determine the age specific prevalence of antihepatitis A antibodies (anti HAV Abs) among children in Delhi. Four hundred and twenty children aged 0-12 years attending outpatient department for vaccination or any minor illness were studied. Sera was tested by ELISA for anti HAV Abs using a commercial kit (Hepvase A 96 TMB). Thirty samples of cord blood were similarly analyzed. All samples of cord blood were positive for anti HAV Abs. Prevalence of anti HAV Abs was 80% by 5 years of age. The most vulnerable age group was 0.5-1.5 years (anti HAV Ab positivity). Cord blood had 100% positivity. Univariate and multivariate analyses taking anti HAV antibody positivity as dependant variable demonstrated that age and father's education (socioeconomic status) significantly affect prevalence of anti HAV Abs. Sex, water supply, history of jaundice in self or family did not have any significant effect on anti HAV antibody positivity. Prevalence of anti HAV antibodies is 80% by 5 years of age. Further studies in different strata of society and different regions in the country are required to assess the need and age for vaccination. PMID- 10832230 TI - Plumbism--a mimicker of common childhood symptoms. AB - Lead poisoning is a preventable entity that can affect almost every system of the body. Its toxic effects range from subtle common childhood symptoms to even death. In a prospective study forty six children with at least one symptom compatible with plumbism were enrolled after screening patients attending medical services at AIIMS. The work-up included a detailed clinical history and examination including intelligence quotient (IQ) and behaviour. Blood lead levels were estimated in all with due precautions. Their ages ranged from 2.5 to 18 years and M:F ratio was 2:1. The common symptoms included unexplained recurrent colic (39), anorexia (21), sporadic vomiting and constipation (13), pigmented gum lines (11), growth failure (11), history of pica (9) etc. None of the children had acceptable blood lead levels (< 10 micrograms/dl). Ninety three per cent children had levels > 20 micrograms/dl and 47.8% had > 45 micrograms/dl. Statistically significant correlation with blood lead levels was seen with most symptoms. Plumbism can mimic common childhood illnesses and should be investigated in children with a clinically compatible profile. Community awareness with regard to pica reduction may be an important preventive measure. This silent epidemic needs to be aggressively tackled by a multipronged approach. PMID- 10832231 TI - Blood lead and erythrocyte protoporphyrin levels in Kazakhstan. AB - A pilot study was conducted to examine the extent of lead exposure and prevalence of iron deficiency in 3 major cities of Kazakhstan. Blood lead (B-Pb.) and erythrocyte protoporphyrin (ZnPP) levels of 475 children, age range 6 months to 7 yeas were measured. The mean B-Pb. levels in the different cities ranged from 4-7 micrograms/dl (minimum 1 to max 29 micrograms/dl) and similarly the mean ZnPP levels ranged from 26-32 micrograms/dl (minimum 12 and maximum 95 micrograms/dl), thus confirming low level lead poisoning of children at some sites. One to four year olds had greater than 10 micrograms/dl B-Pb in 18-27% cases compared with 3 7% cases in five to seven year olds. Prevalence of iron deficiency in 6 months to 4 year old children was the highest ranging from 28-86% compared with 4 to 15% in 4-7 year olds. However, there was remarkably low prevalence (4%) of iron deficiency in a group of 5-6 years olds. This study suggests that a targeted B-Pb and ZnPP monitoring together with an iron supplementation programme in the 3 cities of Kazakhstan is essential. Environmental education appears to have had a positive impact in lowering B-Pb at one site and should thus be expanded nationwide. PMID- 10832232 TI - Adolescent nutrition in a rural community in Bangladesh. AB - The objective of this study was to assess the nutritional status of adolescent boys and girls in a rural community in Bangladesh. Between December 1996 and January 1997, a cross-sectional survey was carried out in 803 households, each containing at least one adolescent, sampled consecutively from four purposely selected villages in Rupganj Thana, Narayanganj district. Initially, the guardians of 1483 healthy and unmarried 10-17 year old adolescents (51% boys and 49% girls) were interviewed about family structure and socio-economic status. Out of these children, 906 (47% boys and 53% girls) from 597 households were weighed, had their height and MUAC measured and were clinically examined. Blood was then collected from 861 adolescents for haemoglobin estimation. The median monthly income per person in these 597 families was approximately Taka 554 (US $12). Twenty seven per cent of the household heads were labourers, 21% were solvent farmers, 14% ran small scale businesses and 6% were unemployed. Sixty seven per cent of adolescents were thin (defined as BMI < 5th centile of WHO recommended reference) with 75% boys and 59% girls being affected. The percentage of thin adolescents fell from 95% at age 10 years to 12% at age 17 years. The prevalence of stunting (height for age < 3rd centile NCHS/WHO) was 48% for both boys and girls and rose from 34% at age 10 to 65% at age 17. On clinical examination angular stomatitis was present in 46%, 27% had glossitis, 38% had pallor, 11% had dental caries, 3.2% had an conspicuously enlarged thyroid and 2.1% had eye changes of vitamin A deficiency. According to INACG (International Nutritional Anaemia Consultative Group, 1985) cut-off values, 94% of the boys and 98% of the girls were anaemic. We conclude that rural Bangladesh adolescents suffer from high rates of malnutrition and almost universal anaemia. Nutritional interventions to target this population are urgently required. PMID- 10832233 TI - Mothers' skills in preparing oral rehydration salt solution. AB - This was a community based thirty cluster survey, carried out in rural Chittagong district, Bangladesh during 1996 with the objective to assess the skill of mothers in preparing packet ORS solution. A total of 420 mothers whose children had been suffering from acute diarrhoea were investigated. There were 377 (89.8%) mothers who demonstrated the preparation of ORS and 43 (10.2%) mothers never ever prepared the solution and were unable to demonstrate the preparation. One hundred and forty (33.3%) mothers were able to demonstrate the preparation correctly and the rest 237 (56.4%) demonstrated the preparation incorrectly. The incorrect preparation was found to be associated significantly with the refusal of ORS solution by the children (p < 0.01). None of the maternal factors were found to be associated with the correctness of preparation of ORS solution except previous exposure of the mother to the demonstration of ORS solution preparation (p < 0.000). Therefore, demonstration of preparation of ORS solution to the mothers should be in built in the health education package of oral rehydration therapy for diarrhoeal diseases. PMID- 10832234 TI - Factors predicting the non-utilisation of immunisation services using logistic regression technique. AB - A cross sectional study was conducted to determine the variables which have an individual predictive power on nonutilization of immunisation services using logistic regression model. Children between 12 and 23 months of age were assessed and enrolled using a pretested interview schedule. Statistical analysis was done using descriptive statistics, univariate analysis using Chi Square tests and multivariate logistic regression analysis. Of the 200 children studied 56% were fully immunised. Only illiterate mothers (p < 0.05) and poor access (> 3 km) to health facility (p < 0.001) were found to have an independent predictive power on the nonutilization of immunisation services. Sex of the child, parity levels, socioeconomic status did not independently influence the utilisation pattern. It is possible to identify these still existing small pockets of nonusers of immunisation services through the predictive variables and target them through special efforts. PMID- 10832235 TI - Cytogenetics investigation in childhood chronic myeloid leukemia. AB - Cytogenetics investigations, mostly from peripheral blood, were carried out in 30 children with CML. Amongst a sample of 30 patients, 18 had chronic myeloid leukemia of adult variety (ACML), while the remaining 12 children had the juvenile type of chronic myeloid leukemia (JCML). Sixteen (88.9%) out of the 18 patients suffering from ACML tested positive for the classical Philadelphia chromosome translocation t(9; 22). Of the remaining two ACML patients, one tested positive for t(9; 13; 22) while no visible chromosomal changes were observed in the other patient. The activity of Nucleolar Organizer Region (NOR) was significantly reduced in 11 (61.1%) of the 18 patients suffering from ACML, when compared to that of 21 normal healthy controls. Ten out of the 12 patients suffering from JCML had normal karyotypes, while monosomy 8 and 21 q deletion were seen in the remaining two patients respectively. Amongst the 30 CML patients, chromosomal abnormalities were observed in 19 patients. Variant Philadelphia chromosome translocation (9; 13; 22) and monosomy B were observed in ACML and JCML, respectively. In two ACML patients, cytogenetic studies were helpful in diagnosis of the disease. PMID- 10832237 TI - Magnesium sulfate in acute asthma. PMID- 10832236 TI - Anti-leukotrienes in asthma: yet to arrive. AB - Inflammation plays a predominant role in the pathogenesis of asthma. The leukotrienes (LTs) exert their actions by binding to and activating various receptors. Leukotrienes B4, C4, D4, and E4 have been shown experimentally to play a role in inflammatory mechanisms, producing the pathologic changes seen in asthma. Antileukotrienes represent a new class of anti-asthma drugs with anti inflammatory role. In asthma management, LT modifiers from the groups of 5 lipoxygenase inhibitor and Cys LT1 receptor antagonists are found useful. LAs are of main use in mild to moderate chronic asthma. Their usefulness is also observed in allergic rhinitis and even in severe chronic cases of asthma which are resistant to steroids. In chronic asthma they are required to be used for prolonged periods with other agents viz. inhaled steroids and beta 2 agonists. These agents are essentially safe. Except for Montelukast, which can be used in children above six years of age, the paediatric use of other agents is yet to be established. LAs are gradually becoming available in increasing number of countries. In India, we have to presumably wait for sometime before these drugs reach the market. The cost of LAs is reasonably high. Thus, India awaits arrival of LAs, may be for good, as more concrete information from various trials will permit us to practice more evidence based medicine. PMID- 10832238 TI - Respiratory illnesses are among the most common causes of morbidity and mortality during childhood. PMID- 10832239 TI - Measuring pulmonary function in infancy. AB - In recent years there has been a growing interest in the measurement of pulmonary function in infants for both clinical and research purposes. Such measurements remain limited by the complexity of the equipment as well as by the technical and physiological challenges of testing infants and neonates. Despite these problems, assessment of respiratory function in early life provides exciting information about the post-natal growth and development of lungs in health and disease. The aim of this paper is to discuss the physiological, technical and ethical problems surrounding these procedures, as well as reviewing the current methods of testing pulmonary function in the very young. Consideration is given to the developments needed if infant pulmonary function tests are to realise fully, their potential as research and clinical tools. PMID- 10832240 TI - Somatizing disorders affecting the respiratory tract. AB - Respiratory disease in childhood is common but not all children presenting to the paediatrician have an underlying organic cause for their symptoms. This article reviews the spectrum of non-organic somatization disorders that might be encountered and advises about the diagnosis and treatment of habit cough, laryngeal dysfunction and hyperventilation. PMID- 10832241 TI - Bronchiectasis: causes and management. AB - Bronchiectasis is a condition representing abnormal and permanent dilatation and distortion of medium sized bronchi, usually accompanied by destruction of the airway wall. Post inflammatory bronchiectasis remains very common in the developing countries as a sequel to pulmonary tuberculosis, whooping cough, and severe measles (among other causes). Cystic fibrosis is the most common cause of generalized bronchiectasis in developed countries. Symptoms primarily are chronic cough and expectoration of foul smelling sputum. Bronchography was, until recently, the investigation of choice for the diagnosis of bronchiectasis and the gold standard against which the current best imaging technique HRCT (high resolution computed tomography) has been compared. Treatment includes prompt attention to acute exacerbations, management of airway secretions and control of airway hyperreactivity. Treatment is aimed at the non progression of the disease and complete cure if possible. The role of surgical therapy has evolved from early curative resection for all patients to a more palliative approach. Patients with advanced generalized bronchiectasis should be considered for lung transplantation. PMID- 10832242 TI - Investigation of the child with interstitial lung disease. AB - Many disorders can affect the pulmonary interstitium in children. Although individual interstitial lung diseases are rare, the range of conditions encountered is wide. Interstitial disease is also seen increasingly as a consequence of the treatment of children having other primary problems including cancer, immunodeficiency and haemotological diseases, as well as in recipients of solid organ and bone marrow transplants. The management and prognosis of individual conditions is highly variable, thus it is essential to search for a precise diagnosis in every patient. High resolution computerised tomography (HRCT) and other less invasive investigations may be helpful in the management of patients. However, it is unusual to be able to make a firm diagnosis without a lung biopsy. PMID- 10832244 TI - Spontaneous rupture of choledochal cyst. AB - Choledochal cyst in an uncommon congenital anomaly with classic presentation triad of abdominal pain, jaundice and right upper abdominal mass. Presentation due to biliary peritonitis following cyst rupture is extremely rare. One such case which was successfully treated is being reported. PMID- 10832243 TI - Asthma--the changing face of drug therapy. AB - Until the recent introduction of long acting beta 2-agonists and the leukotriene antagonists, the drug treatment of asthma had remained largely unchanged for a quarter century. Recent studies have demonstrated the efficacy of the long acting beta 2-agonists in the management of asthma in children and highlighted their value as an adjunct to inhaled corticosteroids. The leukotriene antagonists are an important new class of drug therapy which target a specific area of asthma pathogenesis. Whilst they have been shown to be effective for asthma, their exact role in the clinical situation remains to be established. Recent guidelines have emphasised the important role of inflammation in persistent asthma and recommended the early institution of anti-inflammatory treatment. Many patients remain uncontrolled despite high doses of anti-inflammatory agents including oral corticosteroids. Recent experience with other immunomodulatory agents such as cyclosporin, methotrexate and intravenous immunoglobulin has highlighed their potential as steroid sparing agents. With improved understanding of asthma pathogenesis the potential for specific targeted therapies has become evident. Monoclonal antibodies to IgE and certain cytokines are being investigated as possible treatments for asthma. Similarly, preliminary studies of selective phosphodiesterase inhibitors in asthmatic individuals have been encouraging. Other potential therapies include platelet-activating factor receptor antagonists, tryptase inhibitors and prostaglandin E analogs. The continued development of such targeted treatments should ensure a greater diversity of therapeutic options for the management of asthma in the new millennium. PMID- 10832245 TI - Achalasia cardia. AB - Achalasia cardia is an uncommon condition in children. It needs special consideration as it is associated with a higher incidence of recurrent respiratory infection and failure of growth and development, thereby requiring an early surgical intervention. Four such cases who were successfully managed surgically are described. PMID- 10832246 TI - Congenital methemoglobinemia. AB - A case of congenital methemoglobinemia presenting at birth is reported. The clinical signs, differential diagnosis, pathogenesis and management of this disorder are discussed. PMID- 10832247 TI - Black water fever. PMID- 10832249 TI - The distant dentist. PMID- 10832248 TI - The noble lie. PMID- 10832250 TI - Myofascial pain. PMID- 10832251 TI - Posture training for TMD patients. PMID- 10832252 TI - Fluoride varnishes. A review of their clinical use, cariostatic mechanism, efficacy and safety. AB - BACKGROUND: This is a review of the clinical use, cariostatic mechanism, efficacy, safety and toxicity of fluoride varnishes. TYPES OF STUDIES REVIEWED: The authors reviewed and summarized in vitro, in vivo and in situ studies; clinical trials; demonstration programs; position papers; and editorials published in English in the biomedical literature since 1966. RESULTS: Extensive laboratory research and clinical trials conducted in Europe and elsewhere show that fluoride varnishes are as efficacious as other caries-preventive agents. Fluoride varnishes are widely used in European caries-preventive programs. The U.S. Food and Drug Administration has cleared these products only as medical devices to be used as cavity liners and for the treatment of hypersensitive teeth. These products have not yet been cleared for marketing in the United States as caries-preventive agents. CLINICAL IMPLICATIONS: Three fluoride varnishes are currently available in the United States. Semiannual applications are the most proven treatment regimen. Varnishes are safe and easy to apply and set in contact with intraoral moisture. PMID- 10832253 TI - Effectiveness of sealant treatment over five years in an insured population. AB - BACKGROUND: The authors analyzed an insurance claim database to evaluate the use and effectiveness of placing sealants on first and second permanent molars of children in private dental practices. METHODS: A retrospective study was conducted using an insured population with sealant coverage at 70 percent of the usual fee in a preferred provider organization than that limited reapplication to once every three years. Children were selected who were eligible for sealant benefits from July 1, 1990, to June 30, 1991. Two groups were selected for analysis, based on age, as determined by permanent molar eruption dates. Children who received sealants were compared with those who did not to determine post sealant restorative outcomes after five years. RESULTS: During the full year of coverage, sealants were used in only 16.3 percent of first permanent molars and 11.6 percent of second permanent molars. The five-year incidence of an occlusal restoration having been placed was 13.7 percent and 20.8 percent, respectively, on nonsealed first and second molars and 6.5 percent and 10.4 percent, respectively, on sealed first and second molars. From years three to five, sealant placement provided only nominal additional preventive effect. CONCLUSIONS: In the population selected, both the incidence of occlusal caries and the use of sealants were lower than expected for both age groups. However, within these groups, molar occlusal surfaces were only half as likely to have been restored in sealed teeth than in nonsealed teeth after five years. CLINICAL IMPLICATIONS: Based on the five-year data from a population with a low incidence of caries, the authors found that 15 sealed first permanent molars or 10 sealed second permanent molars prevented placement of one occlusal restoration. Therefore, sealants are more effective when placed in patients with risk factors for occlusal caries. PMID- 10832254 TI - Clinical performance of bonded amalgam restorations at 42 months. AB - BACKGROUND: Despite a lack of data based on clinical research, many positive characteristics have been attributed to the placement of amalgam restorations with an adhesive resin liner. METHODS: For 42 months, and authors followed two groups of subjects who had amalgam restorations placed in a previous study. In this double-blind study, these subjects had been randomly assigned to have amalgam restorations placed with an adhesive liner or with a copal varnish placed under all restorations and a bulk base of zinc phosphate cement for deeper lesions. The authors evaluated anatomical form, marginal adaptation, retention and the presence of secondary caries at six, 18, 30 and 42 months. RESULTS: At 42 months, the authors found that all restorations in both groups still were retained, were free of secondary caries and were rated clinically acceptable. No difference between the groups was found for any category (P > .05; analysis). CONCLUSIONS: Placement of amalgam restorations with adhesive liners was found to produce results equivalent to that of traditional methods over a 42-month period. CLINICAL IMPLICATIONS: Practitioners wary of using new methods that have not undergone thorough clinical testing can feel comfortable placing adhesive liners under amalgam restorations. PMID- 10832256 TI - Treatment options for impacted teeth. AB - BACKGROUND: A compilation of treatment options for impacted teeth is presented to assist dentists in discussing the sequelae of impacted teeth as well as the complications of treatment with their patients. OVERVIEW: A differential diagnosis for an impacted tooth is not possible without clinical assessment. However, an ankylosed tooth or a tooth with failure of its eruption mechanism may be mistaken for an impacted tooth. This review of national and international dental texts, journals and publications concerning impacted teeth spans more than 50 years. PRACTICE IMPLICATIONS: Treatment options for the management of impacted teeth are separated into four categories: observation, intervention, relocation and extraction. The indications, contraindications and complications of each option are presented. This information will help the clinician identify developmental conditions associated with an increased probability of tooth impaction and assess available treatment options. PMID- 10832255 TI - Early orthodontic treatment: what are the imperatives? AB - BACKGROUND: Then authors provide a critical review of the issues involved in determining the appropriate timing of orthodontic treatment. Both single- and two phase treatments are discussed and guidelines are offered to assist in formulating treatment plans. OVERVIEW: In providing orthodontic care for pediatric patients, clinicians often question whether to begin treatment early- during the primary or early-transitional dentition--or wait until all or most of the permanent teeth are present. The authors review the most current literature (from 1991 to 1999), including several recently completed and ongoing randomized clinical trials, to critically evaluate the effectiveness of each approach. PRACTICAL IMPLICATIONS: The controversy surrounding early vs. late orthodontic treatment is often confusing to the dental community. This article reviews both sides of the issue for orthodontic treatment of Class II and III malocclusions, as well as for the management of Class I crowding and problems in the transverse dimension. Early orthodontic treatment is effective and desirable in specific situations. However, the evidence is equally compelling that such an approach is not indicated in many cases for which later, single-phase treatment is more effective. Therefore, clinicians must decide, on a case-by-case basis, when to provide orthodontic treatment. For many patients, delaying treatment until later in their dental and skeletal development may be advisable. PMID- 10832257 TI - Efficacy of articaine: a new amide local anesthetic. AB - BACKGROUND: The authors compared the safety and efficacy of 4 percent articaine with epinephrine 1:100,000 with 2 percent lidocaine with epinephrine 1:100,000. METHODS: In three identical randomized, double-blind, multicenter trials, subjects 4 to 80 years of age received either 4 percent articaine with epinephrine 1:100,000 or 2 percent lidocaine with epinephrine 1:100,000 for simple or complex dental procedures. In each trial, the authors randomized the subjects in a 2:1 ratio to receive articaine or lidocaine. Efficacy was determined by both subject and investigator using a visual analog scale, or VAS. The authors used the Kruskal-Wallis test to analyze the data. RESULTS: A total of 882 subjects received articaine, and 443 received lidocaine. The authors found no statistical differences between the groups (P = .05). They also compared drug volumes for both articaine and lidocaine groups (2.5 milliliters +/- 0.07 standard error of mean, or SEM, vs. 2.6 mL +/- 0.09 SEM for simple procedures and 4.2 mL +/- 0.15 SEM vs. 4.5 mL +/- 0.21 SEM for complex procedures). The procedures' durations were comparable for both the articaine and lidocaine groups. The authors found no statistical difference between the two treatment groups (P = .05) with respect to subject or investigator pain ratings using the VAS; the mean pain scores determined by both patients and investigators for all groups tested were less than 1.0. CONCLUSIONS: The authors found that 4 percent articaine with epinephrine 1:100,000 was well-tolerated in 882 subjects. It also provided clinically effective pain relief during most dental procedures and had a time to onset and duration of anesthesia appropriate for clinical use and comparable to those observed for other commercially available local anesthetics. CLINICAL IMPLICATIONS: Pain control is a major component of patient comfort and safety. Local anesthetics form the backbone of pain control techniques in dentistry. Four percent articaine with epinephrine is an amide local anesthetic that will meet the clinical requirements for pain control of most dental procedures in most patients. PMID- 10832258 TI - Hiring, staffing policies and productivity. AB - BACKGROUND: Many dentists say that staff-related issues, not clinical dentistry, are the number one cause of stress in their offices. OVERVIEW: To help avoid staffing problems and the resultant stress, the author suggests that dentists take the time to identify their specific staffing needs and create job descriptions to prepare for interviewing potential staff members. She provides guidelines for the planning and interviewing processes. PRACTICE IMPLICATIONS: Taking the time to hire the right staff members will help dentists create a productive environment that will benefit them, their staff members and their patients. PMID- 10832259 TI - Survey of U.S. dentists' knowledge and opinions about oral pharyngeal cancer. AB - BACKGROUND: Oral pharyngeal cancer constitutes the most life-threatening of all dental and craniofacial conditions. The U.S. five-year survival rate of 52 percent for these cancers is one of the lowest and has not changed in decades. METHODS: The authors mailed a pretested survey to 7,000 randomly selected general dentists. They obtained information on 3,200 dentists' levels of knowledge about oral pharyngeal cancer risks and diagnostic procedures for providing an oral cancer examination, as well as about related opinions and interest in continuing education, or CE, courses on the topic. The authors carried out analyses using unweighted data; they used both bivariate and logistic analytical techniques and evaluated at a significance level of P < or = .01. RESULTS: Based on responses to 14 questions, the average knowledge of oral cancer risks score was 8.4. About one half of dentists surveyed knew the two most common sites of intraoral cancer and that most oral cancers are diagnosed at a late stage. CONCLUSIONS: The reported knowledge of these dentists regarding oral cancer suggests that they are not as knowledgeable as they could be about cancer prevention and early detection and that they recognize these deficiencies. Most of the dentists were interested in oral cancer CE. CLINICAL IMPLICATIONS: Dentists need to know where in the mouth to look and what types of lesions to look for to provide a comprehensive oral cancer examination. PMID- 10832260 TI - The reinforced composite post and core. PMID- 10832261 TI - Subacute bacterial endocarditis: considerations for the pediatric patient. PMID- 10832262 TI - Prosthodontics is in your future. PMID- 10832263 TI - Obesity, brain and gonadal functions, and osteoporosis. PMID- 10832264 TI - Chiropractic case includes a cautionary for dentistry. PMID- 10832265 TI - [Vegetative state. Practice guideline for its diagnosis and treatment]. AB - Vegetative state (VS) is a clinical condition in which patients have lost both their consciousness content and alertness. So defined, it seems a simple diagnosis to achieve. However, facts disprove this idea, and indeed a striking evidence exists that doctors are unable to satisfactorily diagnose VS. In this series of 3 articles, a multi-disciplinary Italian working group proposes solutions in order to properly diagnose VS (Part I) and to differentiate it from other apparently similar situations (part II). Consciousness, coma and VS state are clearly defined and the reader is provided with easy-to-use methods to achieve the goal. Differential diagnosis includes the locked-in syndrome, the akinetic mutism, and the minimally conscious state, this latter being a challenging situation, which deserves great reader's attention. In the last part the authors afford the delicate argument on when and how a VS can be defined as permanent. Modern research on the use of magnetic resonance imaging in trauma patients and evoked potentials in anoxic patients are presented, and a flow-chart rationalizes the process of outcome prediction. Rehabilitation programs are also indicated to facilitate the continuum of care from the acute to post-acute phase. Lastly, the authors propose the problem of whether interventions are worth doing in such an extremely disabled population. No definite answers are given, and question remains open to discussion. It could not have been that so: doctors cannot be the only judge when a life worth living or not is to be sentenced. PMID- 10832266 TI - Awareness during general anesthesia. Is this any implication for the outcome? PMID- 10832267 TI - [Anesthesia in single and bilateral sequential lung transplantation. Lung Transplantation Group]. AB - BACKGROUND: Anesthesia for lung transplantation: intraoperative complications and long term results. METHODS: 52 patients were scheduled for 16 single lung transplantations (SLT) (9 fibrosis and 7 emphysema) and 36 bilateral sequential lung transplantations (DLT) (4 bronchiectasis, 6 emphysema, 3 fibrosis, 22 cystic fibrosis and 1 pulmonary hypertension). Anesthesia was induced with propofol or midazolam, and fentanyl or alfentanil. As muscle relaxant vecuronium bromide was used. Anesthesia was maintained with isoflurane, fentanyl in boluses or sufentanil continuous infusion in O2 100%. Prostaglandin E1 (20-300 ng/kg/min), inhaled nitric oxide (10-40 ppm), dobutamine (5-15 mcg/kg/min), norepinephrine (0.05-3 mcg/kg/min) and ephedrine (5-10 mg per bolus) were used for hemodynamic management. In 2 patients inhaled areosolized prostacyclin were administered. RESULTS: Mean pulmonary arterial pressure (mPA) and pulmonary vascular resistance (PVRI) increased after pulmonary artery clamping during first lung (mPA: 3347 nel DLT, 3643 nel SLT; PVRI; 375488 nel DLT, 377420 nel SLT) and second lung implantation (mPA: 3746; PVRI: 263553) and decreased after reperfusion of the first (mPA: 4737 nel DLT, 4329 nel SLT; PVRI: 488263 nel DLT, 420233 nel SLT) and the second lung (mPA: 4629; PVRI: 553260). Only in 9 cases (7 DLT and 2 SLT) C-P bypass was used. CONCLUSIONS: With a strong drug support with pulmonary vasodilators, positive inotropic and systemic vasoconstrictor drugs, in most patients we transplanted C-P bypass can be avoided. Intraoperative deaths were not observed. Two years actuarial survival is 65% for DLT and 60% for SLT. PMID- 10832268 TI - [Brachial plexus block. Effect of low interscalenic approach on phrenic nerve paresis]. AB - BACKGROUND: The aim of the paper was to determine the influence of the brachial plexus block on phrenic nerves paralysis in distal side of interscalenic space (between 1/3 medium and 1/3 inferior), with little volume of local anesthetic drug. Comparative, prospective, randomized study. METHODS: Fourty-three patients, ASA I-II, submitted to surgical operation at superior arm, shoulder excluded. The patients were randomized into two groups: group A, 21 patients, were submitted to brachial plexus block by armpit tract; group B, 22 patients, were submitted to brachial plexus block by low interscalenic tract. Ropivacaine 0.75%, 20 ml, was used as local anesthetic. Plexus was localized by ENS and isolated needles 25 G 35 mm or 22 G-50 mm. EXCLUSION CRITERIA: respiratory disease. Effects of phrenic nerves paralysis were evaluated by mean measures of FEV1, FVC, and PEF, with sitting patients, before and after 30 min of the nerve block. Other parameters we obtained were NIBP, CF, ECG, and SaO2. RESULTS: In the B group FVC, FEV1 and PEF, after brachial plexus anesthesia, diminished of 19.39%, 20.8% and 20.7% respectively in comparison with the same parameters measured before local anesthesia. No patients had dyspnea. In group A significant modifications of respiratory parameters were not recorded. Both groups showed stability of cardiocirculatory findings. CONCLUSIONS: Brachial plexus anesthesia by low interscalenic tract, even using little local anesthetic drug volumes, like other supraclavear block techniques determines ipsilateral diaphragm paralysis. PMID- 10832269 TI - [Locoregional anesthesia in inguinal hernia surgery]. AB - BACKGROUND: It is a current opinion that local anesthesia (LA) is the primary choice in surgical treatment of the inguinal region, particularly herniorrhaphy. The LA technique personally used for herniorrhaphy is described: it consists of iliohypogastric, ilioinguinal and genito-femoral nerve blocks, and incision line anesthetic infiltration. METHODS: From January 1998 to April 1999, 95 patients underwent inguinal herniorrhaphy employing LA: 77 (81%) in elective surgery, 18 (19%) in emergency; 2 cases with bilateral hernia (97 total LA procedures). RESULTS: Partial success was obtained in only 8 cases (8.4%), which required an association with a hypnotic drug ("blended anesthesia": propofol or midazolam): there were no cases of conversion to general anesthesia. Specific complications of local anesthetic drugs infiltration developed in 8 cases on 97 LA procedures (8.2%), but none required reoperation: 6 inguinal hematomas, 1 female external genitalia hematoma, 1 hematocele. CONCLUSIONS: In conclusion, it is stressed that LA is the technique of choice in herniorrhaphy and surgery of other inguinal pathologies, associating high success rates, rare complications and rapid dismissal: this allows for easy management of the patients and a very important reduction of sanitary costs. The association of LA-hypnotic drugs (blended anesthesia) represents another important resource, since it avoids general anesthesia in many cases and allows a rapid psychophysical recovery. PMID- 10832270 TI - [Surgical treacheotomy in intensive care]. AB - BACKGROUND: To test validity, safety, costs and healing-outcome of surgical minitracheotomy after cannula removing. METHODS: DESIGN: Opened, perspective. Site: Polyvalent ICU. PATIENTS: 40 patients underwent surgical minitracheotomy, in a year. Traditional surgical technique was adopted. Minitracheotomy was performed by intravenous general anesthesia with curarization and in mechanical ventilation. Intra-operative monitoring: ECG, non-invasive blood pressure and SaO2 by digital pulse oximetry. If a bleeding appeared, hemorrhage was stopped by vessel binding. RESULTS: Technical difficulty was the same in every tracheotomy carried out. Complications appeared in 7 patients. There were 2 infectious cases for stoma, resolved in 7 and 12 days respectively by local disinfection and antibiotic therapy. There were 5 cases with moderate bleeding; in one patient only a surgical procedure was necessary. CONCLUSIONS: Surgical tracheotomy represents a good and safe method in ICU; complications are few, if safety conditions are observed. Tracheotomy made in ICU reduced the use of medical and paramedical staff and operative rooms; unlike percutaneous procedure, in the surgical technique an always the same set after simple sterilization is used. Healing-outcome of surgical tracheotomy, after cannula removing, is the same with both techniques. PMID- 10832271 TI - [Systemic and pulmonary vascular resistance in brain death]. AB - BACKGROUND: Hemodynamic instability is known to affect brain dead subjects and it can be dangerous for the viability of transplantable organs. Aim of the present study was to assess the hemodynamic performance in brain dead subjects, the changes during the legal observation period and the results of therapeutic management. METHODS: The authors evaluated 28 consecutive adult brain dead subjects, all in intensive treatment, controlled ventilation, infusion therapy and/or dopamine administration and continuous direct monitoring of arterial pressure. Ten hemodynamic parameters have been registered by the thermodilution method and the Swann-Ganz catheter. The Legal Committee performed measurements at the beginning (T0) and the end (T6) of the observation period, which lasts 6 hours according to the current law on death certification (Law N. 578/93). RESULTS: Low systemic and pulmonary vascular resistances have been documented in the majority of subjects (75%), both treated only with fluids and with the additional dopamine administration (dosage lower than 10 ug/Kg/min). The above mentioned reduction was similar at the two different monitored times (T0 and T6). CONCLUSIONS: This situation can be ascribed to the destruction of the cerebral vasoactive centers and the consequent hypotension is due to autonomic nervous system dysfunction. Hemodynamic instability must be treated by fluids and inotropic drugs, but they may cause cardiac and respiratory problems, thus it is suggested to use also low doses of vasoconstrictive drugs, provided that cardiac condition allows this therapeutic strategy. PMID- 10832272 TI - [Approach to the patient in vegetative state. Part I: diagnosis]. AB - Coma is the condition in which both components of consciousness, wakefulness and awareness, are lost. Therefore, the comatose patient neither opens the eyes, nor utters comprehensible words or follows commands. In the vegetative state (VS), awareness (including all cognitive function and emotion) is lost, while the vegetative functions, including breathing and sleep-wake cycles (the wakefulness), persist. Usually VS follows a state of coma caused by traumatic or anoxic brain damage. Pathologic brain examination demonstrates extensive subcortical, white matter, bilateral thalamic, and brainstem lesions in various combinations. Bilateral cerebral cortical lesions are rare, and therefore cortex can be "out of action" despite unaltered structure. More rarely, VS is caused by brain developmental malformations in children, or metabolic or degenerative diseases in children and adults. These are the only cases in which patients are not comatose before VS becomes apparent. After 6-8 weeks of coma, patients regain the eye opening. In such a condition, the term coma is no longer appropriate. If awareness is absent, the patient can be defined as in VS. Finally, the VS is defined as persistent (PVS) when it has continued for at least 1 month. The diagnosis of VS and PVS are clinical and require assessment over an adequate period of time and the skills of a multidisciplinary experienced team. Diagnosis is very complex in infants younger than 3 months, and is not applicable to preterm neonates, with the unique exception of infants with anencephaly. Among ancillary diagnostic tools, dynamic PET studies show promise. PMID- 10832273 TI - [Approach to the patient in vegetative state. Part II: differential diagnosis]. AB - A prerequisite to the diagnosis of vegetative state is the exclusion of apparently similar syndromes, in which the patient retains the consciousness partially or even completely. Some syndromes are not separate nosological entities and should be abandoned: the apallic state, the neocortical death, the decerebrate and decorticate state, the alpha-coma, the vigil or prolonged or irreversible coma are among them. Three conditions deserve special consideration. The term locked-in syndrome describes a patient completely paralysed and mute, but fully conscious, and is usually caused by ischemic lesions of the pons. Several variants do exist, either in the causes and site of lesion. Some patients may become paralysed and mute, but conscious because of polyneuropathies, that is in the absence of any lesions of the central nervous system. The akinetic mutism is a rare condition characterised by loss of speech and nearly absent bodily movements. Painful stimulation may cause appropriate withdrawing, and wakefulness and self-awareness may be preserved, but cognitive impairment is usually present. It must be emphasised that this condition can be due to potentially treatable lesions, such as hydrocephalus and craniopharyngioma. The term "minimally responsive" or "minimally conscious" describes severely disabled patients in whom meaningful responses can be demonstrated, although inconstantly. This condition, the true diagnostic challenge, often represents a transition phase of vegetative patients recovering consciousness. Physicians, physiotherapists and patient's relatives should work all together to reach a correct diagnosis, by using current available methods to monitorize the recovery of consciousness. PMID- 10832274 TI - [Approach to the patient in vegetative state. Part III: prognosis]. AB - To define a vegetative state (VS) as permanent is to declare its irreversibility. In 1994 a North-American multidisciplinary task force, by extensively analysing the literature, concluded that the recovery of consciousness from a post traumatic or non-traumatic VS is unlikely after 12 and 3 months respectively. These conclusions did not obtain unanimous consent. The term permanent was in fact inappropriately used to define either the loss of consciousness or of function. Furthermore, patients with traumatic brain injury have been shown to recover the consciousness in a substantial greater percentage (6-7%) than previously appreciated (1.6%). This is hardly compatible with the peremptoriness of the term permanent, which should be used only in case of certainty. Ancillary tests are important in defining the prognosis. Patients in deep coma after an anoxic brain injury can be predicted as having a poor prognosis (death or permanent VS) with 100% specificity within one week of the insult. Magnetic resonance of the brain can predict patients at high risk of permanent VS within 6 8 weeks of a traumatic brain injury. In conclusion, the available evidence does not permit to define with certainty the patients who have irremediably lost their consciousness after a devasting brain insult. However, it seems possible to reliably define the risk of severe disability. Whether or not this knowledge might or should be used to titrate the intensity of therapeutic approach is to be defined. In this respect, it is central the definition of what an "acceptable outcome" is, certainly not an exclusive medical attribution. PMID- 10832275 TI - [Fulminant liver failure caused by antitubercular drugs. Report of a clinical case]. AB - The incidence of tuberculosis has grown seriously in the last ten years and so have the risks due to drug toxicity of Ethambutol, Isoniazid, Pyrazinamide, Rifampicin. One of the questions is whether a careful monitoring of liver function during anti-tubercular chemotherapy could be useful, given that once severe organ toxicity initiates the survival rate remains under 10% if organ transplant is not available. International literature shows a clear prevalence of this event in Asiatic populations which are now well represented in Italy owing to incoming migrations. A case of fulminant hepatitis in a young Chinese man under treatment for TBC arrived at our ICU with a drug-induced acute hepatitis is reported. PMID- 10832276 TI - [Innovative study protocols]. PMID- 10832277 TI - [Exclusive radiotherapy in stage III esophageal cancer]. AB - BACKGROUND: The purpose of the report is to evaluate the role of radiotherapy in the treatment of stage III esophageal carcinoma and to analyze the influence of site, extension, grade of dysphagia and histology on local control and survival. METHODS: Twenty males and 6 females were submitted to external beam therapy with 4-6 MV X-rays and received 60-70 Gy in fractions of 180 cGY to 200 cGy per day, 5 days a week. Radiation therapy technique was two posterior oblique portals and a single anterior field at 100 cm SAD. After 4500 cGy portals were coned down, holding the spinal cord dose below 4500 cGy. RESULTS: Global response to therapy was 73.1%. Median survival was 11 months. The 2-year survival rate was 12.5% in patients with lesions smaller than 5 cm and 5.5% for those with lesions greater than 5 cm. Patients with grade 2 dysphagia had a median survival of 16 months, those with grade 1, 11 months and 2 patients with grade 0, 4 and 9 months. In all patients mild to moderate esophagitis was observed. Two patients developed esophagotracheal fistula. CONCLUSIONS: Exclusively radiotherapy cannot be considered the treatment of choice in III stage patients. Primary chemoradiotherapy may emerge as the treatment of choice for cancer of the esophagus. PMID- 10832278 TI - [Neuroendocrine tumors of the lung]. AB - BACKGROUND: To review the patients treated at our Institute over the past 22 years and to evaluate the behaviour of 14 atypical carcinoids on the basis of Capella's classification (1994), defining the most suitable surgical approach. METHODS: A total of 118 patients undergoing surgery for lung neuroendocrine tumours were reanalyzed on the basis of their anatomic and pathological characteristics, surgical treatment and survival. RESULTS: The 5-year survival rate was 96.5% for typical forms and 74.5% for atypical forms; the 10-year survival rate was 92.4% for typical forms and 49.8% for atypical forms (p < 0.001). Lymph node metastatic involvement was also an important contributing factor for survival (p < 0.05). A review of the series of 19 patients treated over the past 6 years, diagnosed with so-called atypical carcinoid based on Capella's anatomic and pathological classification, showed that 5 were well differentiated neuroendocrine tumours (WDNT), 8 well differentiated neuroendocrine carcinomas (WDNC) and 1 small-cell lung neuroendocrine carcinoma (SLCL). From a surgical point of view, 10 cases underwent demolitive surgery and 9 conservative surgery. The overall actuarial survival rate in this series was 78% at 5 years, 100% for WDNT and 75% for WDNC. The only case of SCLC died after a few months. CONCLUSIONS: The authors conclude that the 5 and 10-year probability of survival are closely linked to the histological type of the lung neuroendocrine tumour and the presence of lymph node metastasis. Capella's new anatomic and pathological classification helps to establish a more precise prognosis for so-called atypical carcinoids. Lastly, given the malignant potential of these tumours, preference should be given to radical exeresis. PMID- 10832279 TI - [Video-thoracoscopic sympathectomy in the treatment of Raynaud's disease and palmar hyperhidrosis]. AB - BACKGROUND AND AIM: Raynaud's syndrome is a clinical entity characterised by episodic vascular spasm, digital ischemia in response to cold or emotional stimuli and hyperhidrosis. Many patients suffering from Raynaud's syndrome are successfully treated using medical therapy alone. Those patients who do not respond to medical treatment undergo surgery but the indications continue to be a source of controversy. A modern approach to thoracic sympathectomy requires a video-assisted technique. The aim of this study is to attempt to use mini invasive type surgery to treat Raynaud's disease and hyperhidrosis in order to evaluate the real efficacy of thoracic sympathectomy in a large number of patients. The results of this method were compared for the two different pathologies in question. METHODS: The methodology used by this study is based on instrumental and clinical tests performed before and after surgery on treated patients using a comparative criterion and with a minimum 5-year follow-up. The pre- and postoperative diagnostic tests were performed by the vascular surgery laboratory and using a C.W. Doppler and a reflected light photoplethysmograph. Capillaroscopy and laboratory evaluations relating to secondary Raynaud's disease were carried out by internist type structures. The patients enrolled in the study responded to the following criteria: primary Raynaud's disease, palmar hyperhidrosis and associated syndromes. The population came from a mixed sociodemographic background, albeit within a strictly regional zone (Sardinia). A total of 42 patients were studied. The surgical technique used consisted of the ablation of thoracic ganglia from the 2nd to the 4th. RESULTS: The results showed a resolution of symptoms in 95% of patients treated for hyperhidrosis, whereas a 50% recidivation rate was observed in patients with Raynaud's disease alone, although symptoms were less intense. The results for Raynaud's disease were more disappointing, but it is important to remember that surgery is the ultimate choice for cases with advanced lesions which do not respond to medical treatment. Under these circumstances, the possibility of halting the evolution of the pathology represents an auspicious achievement. CONCLUSIONS: The authors affirm that mini-invasive surgical treatment of hyperhidrosis was resolutive during a mean follow-up of 3 years. It therefore represents a valid method which causes minimum esthetic damage to the patient and the greatest functional benefit. The postoperative period is short (about 3 days) and free of major complications. There is virtually no post-surgical pain. PMID- 10832280 TI - [Surgical treatment of secondary hyperparathyroidism]. AB - BACKGROUND: Surgical treatment in the patient effected by secondary hyperparathyroidism consists in subtotal parathyroidectomy or total parathyroidectomy plus autotransplantation of parathyroid tissue. METHODS: The results obtained with surgical treatment of 6 patients observed in the years 1995 1996 are analyzed. Two glands were hyperplastic in four patients, 3 in the others. Every patient was submitted to a subtotal parathyroidectomy. RESULTS: Postoperative course was marked by transient hypoparathyroidism in one case. After 18 months of follow-up, no recurrences were observed. CONCLUSIONS: It is pointed out that in case of secondary hyperparathyroidism subtotal parathyroidectomy represents the surgical treatment of choice, according with literature data. Otherwise total parathyroidectomy plus autotransplantation, characterized by a more complex surgical technique, lead to the same results. PMID- 10832281 TI - Three-year follow-up of 62 cirrhotic patients with hepatocellular carcinoma treated with chemoembolization. AB - BACKGROUND: The aim of this study is to present our experience in 62 patients suffering from hepatocellular carcinoma (HCC) treated with transcatheter arterial chemoembolization (TACE) in two years. METHODS: TACE was performed with injection of doxorubicin mixed with lipiodol before embolization with spongostan. This procedure was repeated for 3 cycles almost. Follow-up was performed by US and CT and by assessment of clinical status and biochemical tests. TACE results were assessed comparing size, local spread and TACE technique with patients' survival. The lesion was single in 51 while multiple in 11. In 6 patients the lesion was greater than 5 cm while in 56 less than 5 cm. RESULTS: Overall survival rates were 95.7% at 6 months, 78.3% at 1 year, 46% at 2 years, 40% at 3 years. The best responses were obtained with single lesions smaller than 5 cm and treated with at least 3 cycles of TACE. CONCLUSIONS: We can conclude that TACE is an efficacious therapeutic choice in the HCC patients who cannot undergo surgery. PMID- 10832282 TI - [Recurrent abdominal pain and "chronic appendicitis"]. AB - BACKGROUND: Chronic appendicitis may be the cause of recurrent abdominal pain. This hypothesis is the subject of controversy. The aim is to clarify the possible existence of a chronic inflammation of the appendix by a clinical and histopathologic study. METHODS: The case history and the preoperative symptoms and serum findings of 269 patients with appendectomy have been studied. All the appendices have been histologically examined. Chronic appendicitis was diagnosed when at least two typical histological factors of chronic inflammation were present. The histological findings of the appendices have been correlated with preoperative clinical and serum findings of the patients. 14-46 months after the appendectomy, the patients have been examined. RESULTS: Histological examination revealed 187 cases (69.5%) with acute appendicitis, 44 cases (16.3%) with non disease of appendix and 38 cases (14.2%) with chronic appendicitis. Recurrent abdominal pain and normal leukocyte count were closely correlated (chi 2 = 18.3, p < 0.001; chi 2 = 21.3, p < 0.001 respectively) with diagnosis of chronic appendicitis. 81.8% of 33 patients with chronic appendicitis who underwent follow up had relief of all the symptoms after appendectomy. CONCLUSION: Therefore, the study seems to confirm the existence of a clinico-pathological condition that can be defined as chronic appendicitis, resolvable with appendectomy. PMID- 10832283 TI - [Gasless laparoscopic cholecystectomy. Selective intervention in a high surgical risk patient]. AB - Disadvantages related to CO2 pneumoperitoneum in high risk patients (anesthesiologic classification in III and IV ASA), have led to the development of the abdominal wall retractor, a device designed to facilitate laparoscopic surgery without conventional pneumoperitoneum. A case of a patient with acute cholecystitis, well-compensated liver cirrhosis, and high respiratory and cardiologic risk (ASA III class), submitted to laparoscopic cholecystectomy with gasless technique is reported. PMID- 10832284 TI - Malignant transformation of an adenomyoma of the cardia and malignant Hodgkin gastric lymphoma. An unusual coexistence. AB - An unusual coexistence of a malignant Hodgkin gastric lymphoma of the gastric antrum with a gastric adenocarcinoma within an adenomyoma of the cardia is reported. This is a previously undescribed occurrence. PMID- 10832285 TI - [Pseudomyxoma peritonei with appendiceal and ovarian mucinous cystadenoma. A case report. Correlations with synchronous or metachronous colorectal carcinomas]. AB - Mucocele is a rare pathology which is difficult to diagnose prior to surgery. The clinical symptoms are aspecific. Diagnosis is generally made at the time of surgery. Explorative laparoscopy is not advised owing to the possibility of encouraging metastatic diffusion. Surgery associated with ex tempore freezer tests must be carried out with the greatest caution to avoid rupture of organs and the spread of mucin. Appendectomy and removal of the adnexa is recommended in benign forms, with or without omentectomy. Omentectomy is necessary together with hystero-adnexectomy and right hemicolectomy in malignant forms, as well as lymph node cleaning and removal of all mucin in pseudomyxoma peritonei. PMID- 10832286 TI - [Ileal invaginations caused by Peutz-Jeghers polyposis]. AB - The Peutz-Jeghers syndrome is considered a familial polyposis syndrome. The polyps are of hamartomatous type. The symptomatology is due to the more voluminous polyps which can necrotize, ulcerate, bleed and cause intussusception and intestinal obstruction. A case of ileum-ileal intussusception due to Peutz Jeghers syndrome is reported. The utility of a genealogic research is underlined and the recent results of the genetic research are evaluated. The surgical therapy of Peutz-Jeghers syndrome can have many aims: 1) to remove all big polyps, 2) to avoid the danger of canceration, 3) to prevent the hemorrhagic or occlusive risks and complications. PMID- 10832287 TI - [Biologically borderline neoplasms: epithelial hemangioendothelioma. A case report]. AB - The case of a patient affected by an epithelial hemangioendothelioma of the arm is described. A definitive diagnosis was possible only by histologic examination. The difficulty to reliably predict the biological behaviour of these tumors is emphasized and therefore the necessity of a follow-up of the patient for many years is stressed, even in the case of a histologically benign tumor. PMID- 10832288 TI - [Spiral computerized tomography in the diagnosis of acute occlusion of the abdominal aorta. A case report]. AB - Aim of this report is to present a case of acute occlusion of the abdominal aorta recognized by spiral-CT. After a brief description of the etiology and pathology of abdominal aorta acute occlusion, stress is laid on diagnostic role of spiral CT, based on personal and literature data. The advantage of spiral respect to non spiral CT is the quicker examination time which is very important, mostly in emergencies. On the contrary, the diagnosis of acute abdominal aorta occlusion is not modified by volumetric CT respect to non-spiral and it is based on the absence of aorta contrast enhancement above the occlusion level. PMID- 10832289 TI - [Observations concerning an unusual presentation in a case of intestinal duplication]. AB - Duplications of the alimentary canal are an infrequent pathology and may be localised in any part of the digestive tract. According to reports by several authors, the jejunoileal localisation is the most frequent. The authors report the case of a 13-year-old patient with a duplicate ileal tract which remained asymptomatic until hospitalisation was required, whose symptoms at onset resembled the clinical manifestations of acute appendicitis. PMID- 10832290 TI - [Carcinoid of the appendix. A case report]. AB - A case of a young male operated on for acute appendicitis due to a carcinoid of the base is reported. Since the tumor was infiltrating the resection margin of the appendix, the patient was later treated with a right hemicolectomy. Carcinoid tumor is unusual, but can be encountered several times during the career of a surgeon (1/200-300 appendicectomy). The tumor is more frequent in women (2-4:1), located at the tip of the appendix (62-78%) and has a diameter less than 1 cm in 70-95% of cases. It is more frequently diagnosed incidentally after an operation for acute appendicitis and occasionally during other procedures (colectomy, cholecystectomy, salpingectomy). Liver metastases are rare (< 2%), related to the dimension of the primitive tumor (21-100% when > 2 cm) and can cause a "carcinoid syndrome": flush, diarrhea bronchoconstriction, cardiac valve disease. Diagnosis is made by the pathologist and staging by conventional radiologic procedures (TAC, US), dosage of neuroendocrine mediators such as 24 hours urinary 5-HIAA. Nowadays 111In-octreotide scintigraphy (SRS) has an 86% sensitivity to detect the carcinoid and is useful for staging and for planning a surgical intervention. Simple appendectomy is adequate treatment for appendiceal carcinoids less than 1 cm in diameter. Adequate treatment for tumors greater than 2 cm is right hemicolectomy. A point of controversy is what to do for tumors in the 1 to 2 cm range. It seems that appendectomy alone is sufficient except in those cases when mesoappendiceal invasion is identified. When surgical margins after appendectomy are not free of tumor, additional surgery seems warranted. Carcinoid tumor of the appendix has a good prognosis with a 5-year-survival rate, of 85.9-100%. When liver metastases are encountered octreotide can relieve symptoms and sometimes the progression of the disease. PMID- 10832291 TI - [Ultrasound in trauma (FAST)]. AB - During last 20 years ultrasonography has been widely used in Europe to investigate abdominal trauma. In United States it has not been considered a reliable test especially because of its lack of sensitivity in diagnosing parenchymal injuries. Recently American surgeons reconsidered ultrasonography in trauma and codified its use into FAST (Focused Assessment for the Sonographic examination of the Trauma patients). The goal is to detect hemoperitoneum and pericardial effusion. Advantages are remarkable: ultrasound is cost-effective, fast, non-invasive, can be performed by surgeons even on unstable patients. FAST is now included in ATLS framework for examination of thoraco-abdominal trauma. PMID- 10832292 TI - [Characterization and prognostic value of colorectal adenomas with molecular genetic markers]. AB - AIM: The study analyses the expression of DNA-ploidy, ki-67, PCNA and p-53 and their role as potential markers of the development of colorectal adenomas. METHODS: 34 adenomas of the large intestine were analysed using endoscopic exeresis in 15 males and 13 females with a mean age of 64 years (range 42-80). Flow cytometric analysis was carried out to calculate the DNA-index and immunohistochemical tests were used to evaluate ki-67, PCNA and p-53. Statistical analysis was based on chi 2 test and Student's "t"-test. RESULTS: The DNA index was not statistically correlated with the histotype and grading. Ki-67 was highly positive in 5 adenomas, 2 aneuploids and 3 diploids, but did not show a clear relationship with other growth parameters. On the contrary, p-53 was statistically correlated with the more advanced degenerative state of adenomas, being pathological in 6 cases (30%), all with marked aneuploidy (p < 0.02) and severe dysplasia (p < 0.004). PCNA, which was also pathological (> 40%) in 4 (23.5%) aneuploid adenomas with severe dysplasia (2 villous and 2 tubulo villous), appeared to be significantly correlated with the DNA index (p < 0.005), showing a proliferative index of exceptional clinical importance. CONCLUSIONS: The results of this study show that p-53 and PCNA are two parameters that contribute to the definition of the degenerative risk and allow patients to be selected for constant monitoring, although additional follow-up data are essential to enable the clinical verification of these results. PMID- 10832293 TI - [Abdominal metastasis of cardias adenocarcinoma]. AB - BACKGROUND: The aim of the present study was to evaluate the incidence of abdominal lymph node involvement of adenocarcinoma of the gastric cardia in relationship with the site and depth of tumor invasion. METHODS: From July 1988 to April 1998, 79 patients with adenocarcinoma of the gastric cardia underwent surgical curative resection and D2 lymphadenectomy at the 1st Department of General Surgery of Verona University. Among these 79 patients, 16 had an adenocarcinoma of the distal esophagus (type I), 26 patients had an adenocarcinoma of the anatomic cardia (type II) and 37 had a subcardial adenocarcinoma (type III). The frequency of lymph node involvement in each of the lymph nodes as classified by the JRSGC were analyzed. RESULTS: In type I carcinoma positive lymph nodes occurred in 20% of pT1, 33% of pT2 and 100% of pT3. Positive nodes along the lower half of the stomach were never found. In type II carcinoma positive lymph nodes occurred in 57% of pT1, 86% of pT2 and 83% of pT3. Metastasis along the greater curvature in 18% of advanced cancers were found. In type III carcinoma positive lymph nodes occurred in 83% of pT2, 94% of pT3 and in 100% of pT4. Nodes along greater curvature were involved in 21% of advanced cases and also infrapyloric lymph nodes involved in 13% of cases. The type II and III advanced tumors had involved paraortic lymph node in 33% of cases. CONCLUSIONS: These results suggest that for tumors of the cardia an extended lymphadenectomy is necessary to ensure the removal of all metastatic nodes. PMID- 10832295 TI - [Complete resection of isolated lung metastasis from breast carcinoma results in a strong increase in survival]. AB - BACKGROUND: Surgical treatment of pulmonary resection for metastatic disease has been proven a valuable therapeutic concept for a variety of neoplasms. Concerning breast cancer, at least 25% of patients develop distant metastases. In spite of increasingly sophisticated palliative therapies, the survival of these patients did not appear significantly prolonged during the last 25 years (19-32 months following diagnosis) and about 95% die from metastatic disease. METHODS: Between 1977 and 1997, three epidemiological comparable groups out of a total of 125 patients of our clinic were treated for isolated pulmonary metastasis following breast cancer. Complete data from 96 patients were retrospectively analyzed following stratification to three groups according their surgical therapy: C, complete resection, 28 patients; I, incomplete resection, 34 patients; N, no resection, 34 patients. RESULTS: Comparison of the three therapy methods concerning stage, histology and receptor levels of the primary tumor, number of metastases and the disease free interval (DFI), as well as adjuvant therapy modalities yielded no significant differences. Mean survival of group C was with 79 months (5-years-survival 80%, 10-years-survival 60%) significantly better compared to groups I and N (p < 0.00002). Mean survival of group I and N was not significantly different (15.5 and 9 months respectively). The DFI had no impact on the survival of group C, but showed a high correlation with the survival of group N (R2 = 0.81). CONCLUSIONS: Routine chest X-ray is necessary in the follow up of breast cancer patients and if coin lesions occur, thoracic surgeon should be contacted. PMID- 10832294 TI - [Bronchial carcinoids. Our experience with 35 cases]. AB - BACKGROUND: The aim of this study was to review personal experience regarding bronchial carcinoids. METHODS: This study investigated retrospectively 35 patients with bronchial carcinoids treated in our institution (Unit of General and Thoracic Surgery, University of Bologna) in 20 years (from 1978 to 1997). RESULTS: The m/f rate was 0.6, the average age 42.5 years and the smokers' percentage 42.8. The patients were symptomatic in 88.6% of cases. The carcinoid was located in the right lung in 17 patients (48.5%) and central in 29 patients (82.8%). Surgical treatment included pneumonectomy (6), lobectomy (17), bilobectomy (3), sleeve lobectomy (2), sleeve bilobectomy (1), sleeve resection of main bronchus (1), bronchotomy and tumorectomy (3) and wedge parenchymal resection (2). Thirty patients (85.7%) presented a typical carcinoid and five (14.3%) atypical carcinoid. Peribronchial and/or hilar lymphonodal metastases were present at surgery in 2 cases (5.7%), both centrally located and atypical. The typical carcinoids showed a real 5 years survival rate of 95.8% (with only one death, not related to the neoplasm), while that of the atypical carcinoids was 80% (one patient died of multiple metastases). CONCLUSIONS: The conclusions is drawn that although the carcinoid tumours are a distinct group of neuroendocrine lung neoplasms with a good prognosis in the majority of the cases, lobectomy and sleeve lobectomy are still the standard resection procedure for the majority of carcinoids. For atypical carcinoids lobectomies are the minimal oncologic surgical treatment. PMID- 10832296 TI - [Significance of ultrasonography in the diagnosis of appendicitis]. AB - AIM: The authors aim to evaluate the sensitivity and specificity of abdominal ultrasonography in 183 in-patients (113 females--70 males), aged between 3 and 78 years old, in the General Surgery department of Pugliese Hospital in Catanzaro, for abdominal pain and possible acute appendicitis, using a retrospective study. All patients underwent ultrasonography at the Division of Radiology in the same hospital. METHODS: The technique used was graded-compression US (useful to eliminate gas artifacts and to reduce the distance from the appendix) using a linear transducer between 3.5 and 7.5 MHz. The method lasted an average of 15 minutes and was performed by specially trained radiologists. The transducer was held between the forefinger and thumb and pushed into the abdomen using both palms, as if palpating the abdomen. When compression is applied slowly and gently, the pain is surprisingly well tolerated by the patient. The radiologist records whether the inflamed appendix is visualised ultrasonographically and with what degree of certainty, and whether perforations or the formation of abscesses and other pathological processes can be seen. In this case, clinical diagnosis was confirmed by radiological imaging and eventually by surgical evidence. RESULTS: Of the 183 patients examined, 135 showed positive US findings, 11 refused surgery and pain was resolved by pharmacological treatment, and 9 presented other pathologies (3 gastric ulcers, 4 acute cholecystitis and 2 extrauterine pregnancies). Therefore, 115 patients were effectively positive. Of the 183 patients, 48 were negative but of these, only 39 were effectively negative because 3 were false negatives and 6 revealed other pathologies when examined using other methods of diagnosis. Even if the diagnosis of appendicitis was confirmed by clinical examination in most cases, US is of value both to confirm the clinical diagnosis and to rule out any complications. In this particular case it was also useful for the surgeon as a means of locating the position of the appendix. Even if this method is partly conditioned by the patient's clinical conditions, the results were excellent. CONCLUSIONS: The authors conclude that US of the appendix is a valuable aid in the diagnosis of appendicitis, especially in the case of acute or subacute forms in which other radiological imaging might worsen the pathology and lead to the onset of further complications. US offers undeniable advantages using a non-invasive, low cost technique with a specificity of around 80% and sensitivity between 85 and 93%. It also provides a means of identifying other sources of low abdominal pain. However, we still regard clinical examination as being essential for diagnosis. PMID- 10832297 TI - [The use of the biofragmentable ring (BAR-Valtrac) in colon surgery]. AB - BACKGROUND: Clinical results of colic anastomosis using biofragmentable anastomosis ring (BAR-Valtrac) are presented. Such a method showed to be a real alternative technique to the usual ones. METHODS: Eighty-six colic anastomosis using BAR are collected, 76 of which performed as elective surgery and 10 in emergency. The patients were 47 males and 39 females, with a mean age of 64 years. In 63 cases the patients were affected by colic neoplastic disease, in 16 by complicated diverticular disease (stenosis or perforation) and 7 patients had neoplastic disease of other organs involving the colon BAR device was used in 48 colic reconstructions after segmentary resection and in 38 colic reconstructions after left hemicolectomy. In each case 31-34 mm BAR were used. RESULTS: No perioperative death occurred in our series. Only one case (2%) of anastomotic leak was observed, while in 3 cases (4%) intestinal canalization disorders occurred. No problems for ring expulsion occurred in any patient. Three late complications were observed, as three cases of asymptomatic substenosis discovered during instrumental follow-up and spontaneously cleared up. CONCLUSIONS: On the basis of clinical results, and according to those reported in literature BAR anastomosis is considered a safe, feasible and easy technique to perform colic anastomosis, even in emergency, limited to the intraperitoneal tract of the colon. PMID- 10832298 TI - [Central venous access systems in the oncologic patient]. AB - BACKGROUND: A safe and dependable venous access is mandatory in order to perform cancer chemotherapy and monitor blood values in the neoplastic patient. Prolonged infusions of medications with sclerosing action may damage the vessel wall, inducing chemical thrombophlebitis. Furthermore, extravasation of necrotizing compounds may be a danger to the patient. The application of totally implantable venous access systems (VAS) started in 1982, in the United States of America, where at present 500,000 devices are implanted annually. In Italy such method has been introduced in a later period, with a constantly growing trend. VAS devices have evolved since their first presentation, and so have application techniques: the original surgical route has been supported by the percutaneous one, considered most appropriate by several Centers. In this study, personal experience concerning application of VAS in cancer patients is presented. METHODS: From July 1994 to February 1998, at the General Thoracic Surgery Dept. of the University of Perugia, 198 VAS have been implanted in 195 patients. During the first period all the systems have been applied by surgical cutdown of the cephalic vein (150 cases). In the last 12 month the percutaneous technique for vein puncture has been adopted in 48 patients. RESULTS: Immediate and late complications have occurred: among the former, pneumothorax, hematoma, malposition; among the latter, infection, subclavian vein thrombosis, catheter rupture. The results are analysed after an extensive review of the international literature; pros and cons of the different implantation techniques and the technical aspects useful for preventing complications are underlined. PMID- 10832299 TI - [The diabetic foot seen by the surgeon. Personal experience]. AB - BACKGROUND: The aim of this study is to evaluate a major amputation risk criterion in diabetic patients with trophic lesions of the foot. The records of a series of 100 consecutive patients (65 males and 35 females) with diabetic foot ulcer treated in our surgical facilities between January 1992 and December 1997, in collaboration with diabetologists and podiatrists, have been reviewed retrospectively. METHODS: In 26 cases the ulcer involved both limbs and, therefore, the feet observed in this study have been 126. Accurate diagnosis of the underlying cause was the first step and in cases with a poor blood supply (69 limbs; 55%) unresponsive to medical therapy (44 limbs) vascular reconstruction (37 limbs), spinal cord stimulator (SCS) implantation (3 limbs) or major amputation (4 limbs) were performed. According to Wagner grading there were 42 grade 2 ulcers (33%), 38 grade 3 (30%), 43 grade 4 (34%) ad 3 grade 5 (3%). RESULTS: One patient died postoperatively after SCS implantation. All but 4 neuropathic ulcers (53 limbs) healed in a mean time (+/- SD) of 5.2 +/- 3.8 months and all but 10 vascular ulcers (59 limbs) healed in a mean time of 6.3 +/- 4.1 months. Of the latter group in 4 cases the patient died before ulcer healing while in 6 cases (8.7%) a major amputation was performed (in 2 cases after vascular reconstruction procedures). Minor amputations of the forefoot have been performed in 23 instances (33%) of vascular ulcer and in 10 cases (17%) of neuropathic ulcer. CONCLUSIONS: Since ischemia is the main risk factor for amputation, it is suggested that a particular effort should be made in improving the vascular diagnostic, both clinical and strumental, capabilities of our diabetologists and podiatrists in order to detect the vascular insufficiency in earlier stages. PMID- 10832300 TI - [Malignant tumors of the gastrointestinal autonomic nervous system (GAN tumors or plexosarcomas). Report of a case]. AB - The plexosarcomas or gastrointestinal autonomic nerve tumors (GAN tumors) are very rare neoplasms originating from the gastrointestinal autonomic nervous system. They have been differentiated from the other gastrointestinal stromal tumors due to ultrastructural features showed by electronic microscopy. Their mesenteric localization rare than the gastric and intestinal localization, and it gives a very poor and specific clinical symptomatology. It's very difficult to make a correct diagnosis. The most important prognostic factors influencing the survival seem to be the tumor size and the mitotic activity. The authors describe a case of multiple Plexosarcomas occurred in a young patient one year after a surgical intervention for ileal resection to remove a benign schwannoma. This patient wasn't a carrier of multiple Neurofibromatosis. The diagnostic difficulties before surgical operation are stressed and the importance of surgery as the only diagnostic and therapeutic choice is underlined, even if local recurrences occurred in more than 50% of the treated cases. PMID- 10832301 TI - [Surgical treatment of thyroid microcarcinoma]. AB - Thyroid microcarcinoma is nowaday defined as a tumor of one centimeter or less in the maximum diameter. It occurs in less than five per cent of all thyroid carcinomas. In personal experience four thyroid microcarcinomas have been found in specimens from 121 thyroidectomies and have been retrospectively reviewed clinical data and the long term follow-up of the patients. Total thyroidectomies nor prophylactic lymphadenectomies were never performed. All patients at present are well. Our purpose was to focus on the current surgical treatment of microcarcinoma. Many hypotheses are done to explain pathogenesis and fairly good prognosis of thyroid microcarcinomas, without univocal conclusions. Almost all authors agree that microcarcinoma of the thyroid gland is a low-aggressive tumor, with good biological behaviour, but the therapeutic strategies are not codified yet. Particularly, doubt still exists about the choice between total thyroidectomy and more conservative procedure, and lymphadenectomy, especially when tumor is found in specimens after surgery for benign disease. In conclusion, in our opinion conservative management is the best choice, provided that the patient is included in a correct long-term follow-up. PMID- 10832302 TI - [Liver rupture in HELLP syndrome. Report of a case]. AB - Liver rupture and hemorrhage, are the most unusual and serious complications of HELLP syndrome (hemolysis--elevated liver enzymes--low platelet count). The management should be aggressive by treating coagulopathy and favouring a prompt delivery. Liver rupture remains a surgical emergency with control of bleeding based on trauma surgery. The aggressive approach to the management of these patients led to a remarkable decrease of mortality rates. PMID- 10832303 TI - War injury to the liver. A case report. AB - We report a case of a war thoracoabdominal gunshot wound associated with severe injury to the liver and hemorrhagic shock. Immediately upon admission to the second Echelon Surgical Unit, the patient was operated on with two separate surgical approaches: right thoracotomy and subcostal laparotomy. The patient was transferred to rear hospital for recurrent postoperative hemorrhages. He was eventually transferred to our Department, where he underwent reoperation by the surgical approach through right thoracophrenolaparotomy. Liver lesion debridement and hepatic artery ligation were performed. The early and late postoperative course was normal, and the patient achieved satisfactory recovery on discharge. The case shows that the management of severe liver injuries requires special knowledge and experience from the surgeon. PMID- 10832305 TI - [Necrotizing fasciitis after laparoscopic adhesion surgery. A clinical case]. AB - A case of a fulminating necrotizing cellulitis after a laparoscopic adhesiolysis performed for ileal obstructive disease, is presented. During operation, a very little intestinal perforation was caused by an "atraumatic grasper", and intestinal fluid flooded into the peritoneum; the lesion was sutured. A copious peritoneal lavage was performed and the operation ended by positioning a peritoneal drain. The patient was going well, passing flatus on the second postoperative day; on the fourth postoperative day erythematous edema close one of the trocar wound appeared. In spite of an aggressive medical and surgical therapy the infection had a very rapid spreading and the patient died six days later. A literature review is reported and simple intraoperative remedies are proposed. PMID- 10832304 TI - [Internal abdominal hernia. Unusual cause of intestinal occlusion]. AB - Internal abdominal hernias are an unusual cause of intestinal occlusion. They are responsible for 2% of all the intestinal obstructions. Various types of hernia have been described. The diagnosis is difficult, but should be suspected in patients suffering from intestinal obstruction who have undergone earlier laparotomy. Diagnosis is even more difficult in cases of congenital internal hernia. The authors report a case of left paraduodenal hernia (congenital), and a case of transmesenteric hernia (acquired). Both patients complained of a short history of abdominal pain and characteristic symptoms of acute abdomen (nausea, vomit, cramps and obstipation). Emergency surgery using laparotomy enabled diagnosis and treatment. The authors underline the difficulty of diagnosing these hernias and emphasise the diagnostic and therapeutic importance of emergency surgery. PMID- 10832306 TI - [Malignant nodular hidradenoma. A clinical case]. AB - Malignant clear cell hidradenoma is an uncommon tumor of eccrine sweat gland origin. In contrast to the benign form, malignant clear cell hidradenoma tends to invade the surrounding tissue and has a high incidence of distant metastases. For these reasons a wide surgical excision associated to a locoregional lymph node dissection should be performed alter the initial diagnosis. The case of a 17 year old female with this rare lesion is reported; the literature is reviewed and surgical treatment discussed. PMID- 10832307 TI - [Ventral hernia surgery]. AB - BACKGROUND: A personal technique for polypropilene mesh implantation in the treatment of massive, boundary and recurrent incisional hernias is described. METHODS. DESIGN: retrospective evaluation of cases observed over the last 3 years. SETTING: division of General, Oncological and Thoracic Surgery, Department of Surgical and Anatomical Sciences, University of Palermo. SUBJECTS: twenty-four patients with massive, boundary and recurrent ventral hernia were selected and treated with this technique. INTERVENTIONS: the patients were submitted to implantation of a polypropilene prosthesis by using a part of the well vascularised hernial sac in order to close completely the peritoneal layer under the prosthesis (so as to avoid the contact between prosthesis and viscera) and to close the layer over the prosthesis (avoiding a contact between prosthesis and subcutis). MAIN OUTCOME MEASURES: morbidity and mortality have been evaluated. RESULTS: Patients were discharged from the 7th and 15th postoperative day. No mortality was recorded. In four out of 24 patients the following complications were observed: 1 case of high postoperative fever; 2 cases of superficial infection of the surgical wound; 1 case of wide cutaneous suppuration without prosthesis involvement. No recurrences were observed during the follow-up (6 months-3 year). CONCLUSIONS: Even if a scientific evaluation of the results is not possible due to the poor number of cases, short-term follow-up and unavailable randomized studies, this technique may be useful since it permits to avoid the contact between prosthesis and viscera, with lower postoperative adherences and the isolation of prosthesis from the subcutaneous tissue, sometimes involved in suppuration. PMID- 10832308 TI - Letting go. PMID- 10832309 TI - From cure to comfort. PMID- 10832311 TI - Soul music. Enhancing end-of-life care with music therapy. PMID- 10832310 TI - Telling tales. PMID- 10832312 TI - Learning to be present to dying patients. PMID- 10832313 TI - County-based Medicaid plan sought to improve care. PMID- 10832314 TI - The Minnesota Partnership to improve end-of-life care. PMID- 10832315 TI - Tailoring the message. AB - Health care practitioners can learn how to recognize and respond to patient differences. Applying the techniques outlined in this article can improve the kind and depth of information a patient shares, patient satisfaction, patient follow-through, and actual health outcomes. PMID- 10832316 TI - In search of a good death. The spiritual dimension. PMID- 10832317 TI - Palliative care in Minnesota. PMID- 10832318 TI - Grief and the family. PMID- 10832319 TI - Prescribing opiates for pain relief. PMID- 10832320 TI - Medical education's challenge. PMID- 10832321 TI - Missouri practitioners show notable improvement in diabetic kidney disease screenings. AB - Missouri practitioners participating in a cooperative Diabetic Kidney Disease Project, organized by the Missouri PRO, showed significant improvement in screening for nephropathy among diabetic patients following a quality improvement intervention. This project provides a solid basis for the National Diabetes Project initiated by HCFA as part of the PRO's Sixth Statement of Work. PMID- 10832322 TI - Social Security disability and the physician. PMID- 10832323 TI - Prevalence of autism in Missouri: changing trends and the effect of a comprehensive state autism project. AB - To assess change in autism prevalence in Missouri from 1988 until 1995, computerized client registries from the Regional Diagnostic Centers were analyzed. In the five to nine year age group, the prevalence rose thirty fold from 0.15 to 4.8 per 10,000. The study period coincides with the establishment of the Missouri Autism Project suggesting that provision of services will increase the apparent prevalence figures and that autistic disorders were previously underdiagnosed in Missouri. PMID- 10832324 TI - [Illnesses of the generation. "Temper tantrums on return to the real world"]. PMID- 10832325 TI - [When 5-year-old children complain of migraine... make a media use anamnesis!. Interview by Dr. Beate Schumacher]. PMID- 10832326 TI - [Fit for emergency care service? Test your emergency knowledge on the Pfizer website!]. PMID- 10832327 TI - [ECG with contradictions: what is the site of the infarct?]. PMID- 10832328 TI - [Many indispensible drugs are not studied in pediatrics. Russian pill roulette with children?. Interview by Dr. Stephanie Czajka]. PMID- 10832329 TI - [Restoring osteoporotic bones. Very simple--and usually also quite cost effective]. PMID- 10832330 TI - [Guidelines for therapy of hypertension. One must know them--and apply them on an individual basis]. AB - A target parameter for a cardiovascular risk in patients with hypertension is the lowering of the blood pressure. In 1999, the International Society of Hypertension (ISH) and WHO published new recommendations and redefined normal blood pressure values. When deciding the question what antihypertensive treatment for what patient, concomitant risk factors, hypertension related organ damage and contraindications must all be taken into account. The results of large prospective studies apply only in part for the majority of ambulatory patients. For the effective application of these recommendations and guidelines in the doctor's office, considerably more importance must be attached to the cooperation of all their "users"--including family doctors--when they are being established by a specialist society. PMID- 10832331 TI - [Hypertension and heart failure. Are there approaches for differential therapy?]. AB - Various pathophysiological mechanisms are involved in the development of heart failure. In addition to interstitial changes, restricted myocardial contractility promotes neuroendocrinial activation which, in turn, can lead to structural changes, thus setting up a vicious circle. In order to prevent manifest cardiac insufficiency from developing, individual treatment determined by symptoms and ventricular function and interfering with this circular chain of events must be administered. Incipient cardiac insufficiency is preferentially treated with ACE inhibitors and AT1-antagonists. PMID- 10832332 TI - [Automatic blood pressure monitors. Even with seal of approval they are often inaccurate!]. AB - Owing to the permissible tolerances, even successfully tested automatic blood pressure devices may deliver quite false values. In the largest hypertension study (HOT study) so far carried out, the use of an oscillometric device cannot be considered to provide the "evidence base" for the tacit abandonment of the nonautomatic Korotkoff method, in particular in view of the contradictions in the published data on the device employed. When automatic devices are used in clinical trials, special regulations in the testing protocols of the devices, and a critical consideration of measuring methods in the discussion of the results, are mandatory. PMID- 10832333 TI - [Proper education for the patient! Risk of thromboembolism with oral contraception]. PMID- 10832334 TI - [Are you familiar with the antiphospholipid antibody syndrome? Etiology of thromboembolism in all vascular systems]. PMID- 10832335 TI - [24-hour blood pressure determination. 4. Indications, financial coverage, prospects]. PMID- 10832336 TI - [Myocardial infarct without typical ECG]. PMID- 10832337 TI - [KBV chief Richter-Reichhelm on the new EBM, ICD-10 and physician strikes. Our main goal: removing the budget!. Interview by Jurgen Stoschek]. PMID- 10832338 TI - [Controversy regarding the patient charta. Vote of mistrust against physicians?]. PMID- 10832339 TI - [Tryptan s.c., oral, nasal or as a suppository. Treat migraine on an individual basis]. PMID- 10832340 TI - [Allergy-induced dermatoses. Often cortisone needs a partner]. PMID- 10832341 TI - [Beta blocker therapy for elderly hypertensive patients]. PMID- 10832342 TI - [Thyroid gland and gynecologic diseases. Hope for retinoic acid, selenium and phytosteroids]. PMID- 10832343 TI - [When the stronger sex becomes weak. Testosterone in middle age?]. PMID- 10832344 TI - [Declining testosterone level. When the head wolf becomes a pack animal]. PMID- 10832345 TI - [Erection and strength are measurably improved. When is testosterone substitution indicated?]. PMID- 10832346 TI - [Foot fungus in every 5th sock. Is the patient escaping your awareness?. Interview by Petra Eiden]. PMID- 10832347 TI - [How to deal with recurrent headache pain? Listening carefully may be (almost) enough]. PMID- 10832348 TI - [Smokers with chronic bronchitis and emphysema. Utilize the entire therapy spectrum!]. AB - In the majority of cases, chronic obstructive bronchitis is due to inhalative cigarette smoking. Not only does it cause high costs, but it also has a poor prognosis. The pathogenesis of the illness is now considered to be chronic, mainly neutrophilic, inflammation of the airways and the peripheral parts of the lungs--chronic hypersecretion is a common feature of the disease. Successful treatment requires the giving up of inhalative cigarette smoking. Pharmacological options include anticholinergics, beta sympathomimetics, theophylline and inhalative or systemic corticosteroids. Long-acting beta sympathomimetics such as formoterol, and salmeterol, and still experimental long-acting parasympatholytics, represent significant advances. Such additional measures as vaccination, long-term administration of oxygen and, more recently, surgical procedures such as lung volume reduction to treat severe pulmonary emphysema are described. PMID- 10832349 TI - [A little cough--it is but a minor problem... That can be a dangerous error!]. AB - Although coughing is a useful defensive mechanism, it is, unfortunately, completely non-specific. Almost every pulmonary disease (and a number of extrapulmonary conditions as well) is associated with a cough. In many cases the problem is inflammation to the respiratory mucosae, but also, lamentably, to an increasing extent, a malignant tumour. The commonly heard diagnosis "psychogenic cough" is in reality a real rarity. A 14-point diagnostic program enables a reliable etiological diagnosis to be made in about 90% of the cases, and thus points to the appropriate causal treatment. In all the other cases, there remains antitussive medication--which, however, is still unsatisfactory in severe cases. Of major importance, however, is to ensure that symptomatic treatment with cough inhibitors does not lead to a delay in establishing the diagnosis and thus in initiating essential causal treatment. PMID- 10832350 TI - [Internet pages on the main topic. Chronic obstructive lung diseases]. PMID- 10832351 TI - [When the computer is assisting the surgeon... Navigation systems will also find small brain metastases]. AB - Intra-operative localization systems are increasingly being used in cranial and spinal surgery as orientation aids for the surgeon. The latest of these systems operates without the need for a mechanical or an electrical link between the surgical wound and the computer workstation in the operating room. Since both emission and detection of infrared light are handled by a special camera system, the use of such aids means maximum flexibility for the surgeon. Thus, in contrast to conventional systems, no additional electric cables or mechanical devices that may interfere with the surgeon's freedom of movement, are present. This overview article provides a short description of the technology, and discusses the experience gained with, and the results obtained in, 142 cranial and spinal neurosurgical procedures performed with the aid of the new neuro-navigation system. PMID- 10832352 TI - [Papules on the skin. Morphologically and descriptively oriented dermatologic diagnosis (8)]. PMID- 10832353 TI - [Psoriasis: differential therapy for a common problem]. PMID- 10832354 TI - [10 years psychosomatic quality management. Learn along with your patients!]. PMID- 10832355 TI - [Immediate help for thrombosis. Thrombosis ABC, 11: Fibrinolysis]. PMID- 10832356 TI - [24-hour blood pressure monitoring. 3: Patient preparation, technique and examination follow-up]. PMID- 10832357 TI - [Traveler to the tropics with exanthema and fever. Dengue fever]. PMID- 10832358 TI - [Caution: specialists on a man-hunt. Not to the primary physician first, but immediately to the urologist!]. PMID- 10832359 TI - [Growth factor for diabetic foot ulcer. How man amputations are preventable]. PMID- 10832360 TI - [Stroke prevention. Is there an added value to RAS inhibition?]. PMID- 10832361 TI - Walter J. Kahn, MD. President, the Medical Society of New Jersey. PMID- 10832362 TI - Comprehensive health care review is legislative priority. PMID- 10832363 TI - Gasless versus conventional laparoscopy. AB - The objective of our study was to evaluate the technical advantages of using the Laparolift device and examine the clinical benefits of gynecologic laparoscopy without pneumoperitoneum. A randomized single-blinded clinical trial was performed employing a sample of 30 consecutive patients undergoing laparoscopic gynecologic surgery with a primary outcome measure of visualization. Visualization was found to be inferior in gasless patients. The conclusion was that conventional laparoscopy employing pneumoperitoneum provides surgical exposure superior to that offered by the Laparolift device. PMID- 10832364 TI - Prevention and early diagnosis of cutaneous melanoma. PMID- 10832365 TI - Accounts receivable systems: the key to financial health. AB - Just as medical technology has made quantum leaps during the past few decades, the business side of running and maintaining a medical practice has changed dramatically as well. Gone are the days when physicians could submit bills to insurance companies and expect to receive timely payment. Even longer gone are the days when patients walked into a doctor's office actually prepared to write a check or leave cash for services. Today, the task of billing and collecting is more complex, more time consuming, and more deserving of the attention of a skilled office worker. PMID- 10832366 TI - Treating mental illness--then and now. PMID- 10832367 TI - Planning the end of life--plain talk makes good sense. PMID- 10832368 TI - Investigation of child abuse reports--the law must be served and our citizens protected. PMID- 10832369 TI - Completing the circle. PMID- 10832370 TI - Bottled vs. tap water--how about fluoride and iodine? PMID- 10832371 TI - The Hippocratic Oath. Searching for an ideal. PMID- 10832372 TI - Ancient wisdom for modern medicine. The tradition of Judaism. PMID- 10832373 TI - Early women physicians of eastern North Carolina. Lula Disosway, MD, Malene Grant Irons, MD, and Isa Grant, MD. PMID- 10832374 TI - Bioterrorism. A new threat with psychological and social sequelae. PMID- 10832375 TI - Annual report of the State Health Director. PMID- 10832376 TI - Bright young stars. Remembering some student medical researchers at UNC. PMID- 10832377 TI - Social anxiety disorder. A guide for primary care physicians. AB - Social anxiety disorder is prevalent, potentially disabling, but quite treatable. A thorough and directed history can distinguish social phobia from depression, panic disorder, and OCD. It can also screen for and identify possible substance abuse. Once the diagnosis is made, a combination of pharmacologic and psychotherapy is indicated. The SSRIs, MAOIs, benzodiazepines, and beta-blockers- as well as CBT--can effectively treat social anxiety symptoms. Primary care physicians may well want to begin by prescribing an SSRI like paroxetine, along with a high potency benzodiazepine to be taken on a regular or an as-needed basis, and a beta-blocker to take as needed in anticipation of stressful social situations. A referral for CBT should be considered. If the patient has marked side effects from drug treatment or a lack of adequate response to medication, psychiatric referral is definitely indicated. PMID- 10832378 TI - [Etiology of eye diseases, their prevention with special consideration of risk factors]. AB - Based on the newest data of literary, sometimes on different handbooks and his own more than 40 years experience, the author summarizes the reasons for developments of the eye diseases and the most often occurring endogen and exogen factors. The summary is justified, because both in diagnosis and in therapy, but especially in prevention the closest interdisciplinary cooperation is necessary. The listed information has special importance in everyday medical practice, because a practitioner experiences eye diseases very often. He must know what kind of diseases, which factors can cause eye alterations. It's known that these could be connected with other sicknesses and number of risk factors. The symptoms can often precede the alterations connected with the basis illness (diabetes, hypertonia, etc.). The not recognized, neglected cases can cause serious sight disorders, sometimes can lead to blindness, which results in serious ethical, economic problems for the patient, the family and society. Based on the newest WHO data the distribution of eye-diseases will be published. PMID- 10832379 TI - [Open-heart surgery in Jehovah's witnesses]. AB - The religious community Jehovah's Witnesses was founded in 1870. They hold that blood transfusion is against God's law. Surgical treatment of a Jehova witness is a great challenge for every surgeon, especially for cardiac surgeons because blood transfusion is frequently needed during such operations. Authors have been trying to reduce the utilization of preserved blood for ten years. This study is about the experience with Jehovah's Witnesses who have undergone open heart surgery in Debrecen. Twenty-four patients underwent open heart surgery from 1989 till May 1999. 7 of them were males and 17 were females. The mean age was 53 years (40-70 yrs). Three patients had congenital heart disease, 11 had acquired valve disease and 7 had coronary stenosis. In 3 cases the patients had combined coronary and valve disease. Authors used a complete procedure for reducing blood loss during the operations. Two patients (8.3%) died during the early postoperative period. Preoperative mean haemoglobin level was 134.2 g/l (112-166) and haematocrit value varied between 36-50% (mean 38%). On the first postoperative day significant decrease was registrated in these values. From the second day a slow but significant increase of haemoglobin and haematocrit levels were detected. The mean follow up time was 37.6 months (2-144), and the NYHA classification of 21 longtime survivors improved from 3.06 to 1.62. At the Department of Cardiac Surgery in Debrecen as well as worldwide more and more operations are done without blood or preserved blood products, so it could be said that nowadays surgical treatment of Jehovah's Witnesses has lower risk than before. PMID- 10832380 TI - [Activity of neutral endopeptidase in serum of patients with primary biliary cirrhosis]. AB - Neutral endopeptidase (neprilysin; EC 3.4.24.11) is present in the brush border membrane, for example in the bile ducts. We investigated serum neprilysin activity and its correlation with cholestatic markers in patients with primary biliary cirrhosis. Sera of 39 patients with primary biliary cirrhosis (37 females, 2 males, mean age 45 years, range 24-71 years) were investigated. Twenty seven healthy volunteer subjects served as control. Serum neprilysin activity was measured by a sensitive microplate-based continuous monitoring kinetic assay. Succinyl-alanyl-alanyl-phenyl-alanyl-4- nitroanilide was used as substrate. For statistical analysis Kruskal-Wallis ANOVA by ranks and Mann-Whitney U test were used. The neprilysin activities were significantly higher in stages III (mean 13.2 +/- SD 10.8 U/l) and IV (21.8 +/- 17.5) than in the control subjects (2.4 +/ 2.9, p < 0.01). There was no significant difference in neprilysin activity between the patients with stages I and II, or between stage I + II (2.88 +/- 3.0) and the control. Positive correlation was found between the activity of neprilysin and serum bilirubin, alkaline-phosphatase and gamma-glutamyl transferase (p < 0.005 for each). In this study we confirmed that serum neprilysin activity is elevated in patients with primary biliary cirrhosis at advanced stages and the elevation correlated with the cholestatic markers. The increased neprilysin activity seems to be an indicator for the severity and progression of the disease. PMID- 10832381 TI - [Eosinophil cystitis]. AB - Authors report on two rare cases of eosinophilic cystitis, giving a review of the etiologic assumptions and pathogenetic, pathologic aspects of the disease, based on the available literary data. The course of the disease can be either acute or subacute, and is most often chronic. Relapse and progression can interchange in irregular manner, attention is therefore called to the importance of follow-ups. Resection deep into the intact--also containing muscle fibre--as well as histologic examination are considered essential as the only method of differentiation, giving precise diagnosis in the present two cases, too. PMID- 10832382 TI - [Dr. Ern"o Pet"o (1886-1959), the forgotten founder of the community hospital in Szombathely]. PMID- 10832383 TI - [Dr. Emil Zuckerkandl (1849-1910)]. PMID- 10832384 TI - [Treatment of asthma. 1900]. PMID- 10832385 TI - [Collective effort of homosexual men in their fight against AIDS]. PMID- 10832386 TI - [The measurement of inequalities in age of death: calculating the Gini Index based on mortality tables]. AB - The Gini index has been used to gauge the concentration of different variables, including income, the mortality distribution and the spread of physicians. The use of the Gini index for gauging the inequalities in the age of death based on actuarial mortality data. For this purpose, the 1990 age and gender-related mortality figures for each Autonomous Community were used. The inequalities in the number of years lived are greater among males than among females throughout all of the Autonomous Communities. The Communities showing the longest life expectancies are those which have also been revealed to involved the least degree of inequality in the number of years lived, which therefore makes in possible to pinpoint health-care priorities in some cases. The Gini index can be applied to age-related mortality for gauging the degree of inequality in the age of death and to pinpoint age groups on which to place priority with regard to health-care measures. PMID- 10832387 TI - [Women and tobacco: gender-related characteristics]. AB - The percentage of women smokers is still on the rise in Spain. If the current smoking patterns among women continue, the number of deaths will continue rising sharply over the next hundred years. There may be more female smokers than male smokers in the adult population in Spain in the near future. This review provides an update of current knowledge as to the impact cigarette smoking has on women's health, the current patterns of use among them, the special factors which have a baring on their smoking and the difficulties they have when they are considering the possibility of or trying to quit smoking. We also suggest ways in which a minimum degree of counseling, behavior therapy and nicotine substitute can be incorporated as treatment strategies suited to the special needs of women smokers. PMID- 10832388 TI - [High-risk sex behavior and HIV prevalence among gay and bisexual men in the community of Madrid]. AB - BACKGROUND: To analyze high-risk sexual behavior as regards HIV, the use of preventive measures and the patient-reported prevalence of HIV infections among males belonging to one of the leading homosexual associations in the Region of Madrid. METHODS: Cross-sectional study conducted in 1997-1998 by way of mailed anonymous questionnaires. An analysis is made of the sociodemographic characteristics, how often condoms are used for different types of sexual intercourse with regular or casual partners, patient-reported prevalence of HIV and other related aspects. RESULTS: 157 questionnaires were returned by gay/bisexual males. These subjects averaged 32 years of age, 85% having a high school or college education, over the past 3 months, 56% had had intercourse with more than one man; 70.6% practiced insertive anal intercourse with a regular partner and 57.4% with casual partners, solely 32.5% and 61.1% of whom always used a condom. 69.7% had receptive anal intercourse with a regular partner and 39.4% with casual partners, 35.5% and 78.4% of whom respectively always used a condom. 86.6% had oral-genital intercourse, less than 10% having always used a condom. 137 were aware of their serological condition, and 15.2% were HIV positive. 10% had had some STD at some point during the previous year. CONCLUSIONS: A major percentage of those surveyed were involved in high-risk practices (several partners and unprotected high-risk sexual intercourse) which, in conjunction with the major prevalence of infection, can be said to be the same as a major seroconversion rate. PMID- 10832389 TI - [Descriptive study of statistical methods in the original article published on the cigarette smoking habit in four Spanish medical journals (1985-1996)]. AB - BACKGROUND: Being the tobacco use a high-priority subject of investigation and having itself increased the utilization of statistical techniques in biomedical publication the used statistical techniques are described and the statistical accessibility is quantified in the original articles on tobacco use published in four Spanish medical journals. METHODS: Retrospective descriptive study of 154 original articles on the cigarette smoking habit published in 1985-1996 in the journals Atencion Primaria, Medicina Clinica (Barcelona), Revista Espanola de Salud Publica and Revista Clinica Espanola. An only observer codified the statistic techniques in 14 categories in agreement with the classification processed by Carre et al (1995) from the classification settled down by Emerson and Colditz (1983). The knowledge of bivariable techniques, to simple lineal regression, was stablished as the reference for the study of statistical accessibility. RESULTS: 81.8% original articles used inferential statistics. The most frequently used categories were "Contingency tables" (37.0%), "Descriptive statistics" (18.2%) and "Life tables and analysis of survival" (9.7%). A reader familiarized with bivariable techniques has statistical access to 96.0% for the originals of Revista Espanola de Salud Publica, 86.2% of Atencion Primaria, 66.7% of Medicina Clinia (Barcelona) and 33.3% of Revista Clinica Espanola. The same reader had statistical access to 100% for the originals published from 1985 to 1987 and 68.1% from 1994 to 1996. CONCLUSIONS: The use of statistical methods depends on the investigation subject and design, the journal and the year of the publication. The decrease of the statistical accessibility recommends to identify the profile of the standard reader in Spain, to adjust his knowledge to the current biomedical literature demand. PMID- 10832390 TI - [Incidence, etiology and epidemiology of brucellosis in a rural area of the province of Lleida]. AB - BACKGROUND: This a prospective study of the incidence, etiology and epidemiological profile of human brucellosis in the regions of Pallars Jussa y Sobira (Lleida) for the 1995-1998 period. METHODS: Fifty-five patients diagnosed as having brucellosis were studied. Information was recorded regarding the gender, age, town where residing, occupational hazard, contact with animals and intake of unsterilized dairy products, blood samples having been taken for blood cultures. RESULTS: A total of ten cases were reported in 1995, fourteen in 1996, fifteen in 1997 and sixteen in 1998, the average cumulative rates being 52 in Pallars Jussa and 129 in Pallars Sobira. Four times more cases were reported among males (81.8%) than among females (18.2%) (RR: 4.4; CI95% 2.2-8.7). The largest number of cases occurred in March-April, and the fewest during the summer months. Seventy-one percent (71%) of these patients were working at an occupation involving this risk, the direct contagion mechanism being clearly prevalent (71%). The animal species most frequently considered to be the source of infection was that of sheep (65%), followed by cows (47%) and goats (25%). In Pallars Jussa, mainly sheep (RO: 0.3 CI95% 0.1-0.9) and in Pallars Sobira, cows (RO: 6.6; CI95% 1.8-26.2). Twenty-seven strains of Brucella sp, all of the melitensis species, were isolated. CONCLUSIONS: The number of cases of brucellosis in the regions studied have risen in the 1995-1998 period. The results of study of this are indicative of the characteristic profile of an occupational disease. The etiological agent was Brucella melitensis, biovariety 1 clearly being the most prevalent. PMID- 10832391 TI - [Severe acute intoxication treated in the intensive care unit in 1986-1997]. AB - BACKGROUND: To ascertain the epidemiology of the severe acute intoxications in an intensive care unit and to assess the PCR prognosis and death rate related to the different toxic substances. METHODS: Retrospective study conducted in the 10-bed polyvalent intensive care unit at a General Hospital for adult care. Study conducted over a 12-year period. Review of the clinical histories of the patients admitted as a result of severe acute intoxications. Data was gathered regarding demographic aspects. PCR at admission, need to VM, complications of the severe acute intoxications and death rate for the series. An overall as well as a year by-year analysis was conducted. The data was processed statistically employing the SPSS package using the Student "t" or the Chi-square, taking values as being significant if p < 0.05. RESULTS: 233 subjects, 130 of whom were males, were included in this study. The average stay totaled 4 days. Sixty-three percent (63%) of the patients were under 40 years of age (p < 0.05). The most frequent type of intoxication was that caused by one single medication (72%). The survival rate following the PCR was 40% (4/10). The overall death rate totaled 5.6% (n = 13), 92% of the subjects who later died having been required VM at some point in time during their stay in the intensive care unit. CONCLUSIONS: In our environment, the most frequent type of intoxication is that which is caused by medication. The death rate has been revealed to depend upon the intentional aspect, however as being independent of the type of toxic substance (medication or non-medication). The PCR related to severe acute intoxication has a better prognosis for our series than that related to other diseases and disorders. The VM for severe acute intoxications has a low death rate (15.7%). PMID- 10832392 TI - [Set of indicators for the assessment of the degree to which drug therapy guide is followed in primary care]. AB - BACKGROUND: The constant gauging of drug prescription quality is part of the health-care process quality improvement programs as regards Primary Care, but generally speaking, there are no tried and true indicators which can be used in this assessment process. Measuring the degree to which certain set drugs are adhered to as a district-wide guide regarding which a consensus of opinion has been reached can be used as an overall indicator of prescribing-related quality. The objective of this study is that of setting out a method for selecting quality indicators of generalizing type, the same serving as an example with regard to selecting an indicator for gauging the degree to which the district guide is followed. METHODS: Descriptive cross-sectional study entailing the general practitioners in the Health Care District of Aviles (n = 78) as the individual district analyzed, and the prescriptions written thereby which were invoiced in 1997 as the data source. Seven (7) indicators were prepared for gauging the degree to which the District Drug Therapy Guide was followed. A selection was made by studying the following aspects: reliability, ready availability, accuracy, sensitivity, specificity and coherence. RESULTS: The scores fell within the 10-23 range (out of a possible 28), the highest score being that of the percentage of prescriptions for active ingredients out of the guide in comparison to the total number of prescriptions written by each physician. CONCLUSIONS: The indicators used in assessing drug-prescribing quality must first be analyzed in detail before being implemented. An innovative method is suggested for testing indicators for monitoring the prescribing of drugs as a whole. The most useful of all of the indicators studied for gauging the degree to which the Primary Care drug therapy guide was being followed is the percentage of prescriptions written for active ingredients out of the total number of prescriptions written. The degree to which the District Drug Therapy Guide is followed is related positively to quality prescribing. PMID- 10832393 TI - [Latest progress in maxillofacial X-ray study and surgery in treatment of congenital and acquired deformities of viscera cranium in adults]. AB - The communication outlines the present-day potentialities of X-ray diagnosis and maxillofacial surgery in recognizing and treating congenital and acquired deformities of the visceral cranium. PMID- 10832394 TI - [Computed tomography and magnetic resonance imaging in the diagnosis of accessory nasal sinuses, upper jaw and nasal cavity tumors]. AB - Radiation studies, including computed tomography (CT) and magnetic resonance imaging (MRI), revealed that these techniques can accurately determine the site of a tumor, the borders of its spread to the adjacent anatomic structures. They also revealed the features of CT in detecting osseous structural destruction and the advantage of MRI in visualizing the soft tissue component of a neoplasm and in distinguishing the degree of contrast of tumor tissue and concurrent secondary inflammation. The accuracy of CT and MRI for small tumors is 45-80 and 29% higher than that of X-ray study and traditional tomography, respectively. The potentialities of all radiation diagnostic techniques for over 3.0-cm tumors are equal. PMID- 10832395 TI - [Periapical destructive processes of jaw bones and dynamics of osseous tissue recovery after current surgical treatment]. AB - The paper discusses whether periradical destructive changes can be identified at X-ray study and how long it takes to recovery osseous tissues after surgery for these lesions. PMID- 10832396 TI - [Gated 99mTC-MIBI SPECT in assessment of left ventricle global contractility]. AB - The complex radiodiagnostic study (synchronized 99mTc-MIBI SPECT and equilibrum ventriculography) has been made in 30 pts. The opportunity of use synchronized tomoscintigraphy for an estimation of a global left ventricle ejection fraction was shown. PMID- 10832397 TI - [Magnetic resonance imaging in the diagnosis of gastric cancer: X-ray versus MRI anatomic findings]. AB - The paper assesses the present-day role of MRI in the diagnosis of gastric cancer. The authors consider the major prerequisites for the main aim of their study to be: 1) a dramatic incidence of diffuse (endophytic) gastric carcinoma, which requires significant correction of today's approaches to its diagnosis and 2) a rather biased and, in the authors' opinion, present-day mainly negative attitude towards MRI of the stomach as a diagnostic method for its tumor lesions. By applying the X-ray-MRI anatomic principle to the comparative study of MRI findings in 50 patients with predominantly gastric intramural carcinoma and in 25 patients without gastric tumors (controls), the authors present their methods for gastric MRI, the MRI semiotics of gastric cancer by concurrently touching upon a variety of problems that characterize the potentialities of MRI of the stomach in the diagnosis of its tumor lesions, including their differential diagnosis. As a result, the authors highly appreciate gastric MRI and consider this method to be included into the diagnostic algorithm of radiation techniques used in the diagnosis of gastric cancer, which should occupy its definite diagnostic place. PMID- 10832398 TI - [X-ray guided endovascular repair of venous bed of contracted kidney from pyelonephritis in the treatment of renal hypertension and chronic renal failure]. AB - For compensation for chronic ischemia of the pyelonephritically contracted kidney, the authors used X-ray endovascular venous revascularization for the first time. The surgical intervention was to stenose the subsegmental veins of the diseased kidney, resulting in recanalization of the arterial system. The outcomes of X-ray treatment were analyzed in 38 patients with chronic degree I-II renal failure and nephrogenic hypertension. In 35 (92.1%) patients of them, there was improvement in the clinical picture of the underlying disease and in the filtrating and reabsorbing function of the kidney operated on, an increase in its sizes, and a decrease in systemic blood pressure. PMID- 10832399 TI - [Direct needle embolization of hepatic arterial pseudoaneurysm]. PMID- 10832400 TI - [Present-day technical support of mammography]. PMID- 10832401 TI - [The century's results in development of medical roentgen technics]. PMID- 10832402 TI - [Electronic history case register system in Russian Radiology Research Center]. PMID- 10832403 TI - [Diagnostic efficiency and safety of magnetic resonance angiography (review of literature)]. PMID- 10832405 TI - [Nitric oxide in biomedical research]. PMID- 10832404 TI - [May 26, 1999 resolution on the joint meeting of the Board of Radiation Diagnosis and Radiation Therapy Section, Academic Council, Ministry of Health of Russia, the X-ray Diagnosis Ad Hoc Commission, Interdepartmental Scientific Council for Medical Radiology and Radiation Medicine, Russian Academy of Medical Sciences and the Executive Committee, Russian Association of Radiologists]. PMID- 10832406 TI - [The role of nitric oxide and other free radicals in ischemic brain pathology]. AB - Nitric oxide (NO), an intercellular messenger and a normal metabolic product, takes an active part in the regulation of physiologically significant functions of the cardiovascular, immune, and nervous systems. At the same time when produced in excess amounts, NO as a free radical and an agent that gives rise to highly toxic oxidants (peroxynitrile, nitric dioxide, nitron ion), becomes a cause of neuronal damage and death in some brain lesions (parkinsonism, Alzheimer's disease, Huntington's chorea). Numerous experimental data show the ambiguous effects of NO on the development of cerebral infarct. NO as an active vasodilatory and antithrombogenic agent may reduce cerebral damage in early ischemia. There is evidence for the involvement of NO in the body's adaptation to oxygen starvation and ischemic tolerance formation. In the postischemic period, NO is a major factor of neuronal necrosis and apoptosis. The currently established ideas on the processes of cerebral NO production and on the pathogenetic mechanisms of this agent's cytotoxicity open up new vistas for selective blockers of various NO synthesis enzymes (neuronal, endothelial, glial cellular, and macrophagal and neutrophilic NO synthases) used in the treatment of acute vascular abnormalities of the central nervous system. PMID- 10832407 TI - [The role of nitric oxide in brain glutaminergic pathology]. AB - Nitric oxide (NO) is highly reactive signal molecule that is widely distributed in the body, and in the mammalian brain in particular. The agent is a gaseous chemical messenger that acts as a universal modulator of a diversity of physiological functions, such as interneuronal communications, synaptic plasticity, memory formation, receptor functions, intracellular signal transmission, mediator release. NO is assumed to be a key pathological agent due to its neurotoxic effect in developing neurological disorders, such as ischemia, cerebral stroke, epileptiform seizures. The agent is shown to be significantly increased in the brain of rats with abnormalities. PMID- 10832408 TI - [Nitric oxide in the cardiovascular system: its role in adaptive protection]. AB - Nitric oxide (NO) is involved virtually in all processes occurring in the body. In the cardiovascular system, it regulates cardiac contractility, blood coagulability, cell proliferation, vascular tone, and blood pressure (BP). NO deficiency leads to the development of severe cardiovascular diseases associated with endothelial dysfunction, abnormally increased vascular tone and BP (such as hypertension, angina pectoris, atherosclerosis, diabetic angiopathy, etc.) At the same time, NO hyperproduction makes a contribution to the development of septic, cardiogenic, thermal, and other shocks. The most effective non-drug stimulation of NO synthesis is a gradual adaptation to environmental factors. The adaptation induced elevation of NO levels, followed by vascular wall NO deposition, on the one hand, limits the damaging action of excess NO in shocks by the feedback, and, on the other, makes up some NO reserve which may be used in NO deficiency. Further studies of NO-dependent adaptive mechanisms will allow the agent to be used in the prevention and treatment of cardiovascular diseases associated with NO metabolic disturbances. PMID- 10832409 TI - [Nitrogen monoxide in the mechanisms of cardiovascular functional resistance to emotional stress]. AB - The paper presents the results of examination of the mechanisms of development of different physiological dysfunctions (diminished cardiac electrical stability, fibrillation threshold fall) under emotional stress. Search was made for their drug correction (nitric oxide precursor L-arginine, the NO synthase blocker Ng methyl-L-arginine and sodium nitrate). Experiments have indicated that L-arginine and sodium nitrate restore cardiac electrical stability. The normalizing effects of NO donors depended on the stage of cardiac disorders. PMID- 10832410 TI - [The activation of soluble guanylate cyclase by new NO donors as the basis of a directed search for new efficacious vasodilators and antiaggregants]. AB - The paper deals with the molecular mechanisms of antihypertensive and antiaggregatory actions of nitric oxide (NO). These effects of NO are shown to be directly associated with the activation of soluble guanylate cyclase and with the accumulation of 3',5'-guanosine monophosphate. The paper discusses the mechanism of activation of guanylate cyclase with new donors: the role and value of a nitrosyl-hemic complex formed in the interaction of guanylate cyclase with NO. New approaches to studying the mechanism of antihypertensive and antiaggregatory actions of the NO donors in question provided fundamental and priority data that lead to the development of the molecular bases of goal-oriented search and the synthesis of new effective vasodilators and antiaggregants. Examining the molecular mechanisms of targeted activation of soluble guanylate cyclase with new NO donors has revealed new enzyme activators that generate NO in the body and participate in the regulation of homeostasis and vascular tone. PMID- 10832411 TI - [Normal NO synthases and in pathology of different origins]. AB - The paper deals with the physicochemical properties of nitric oxide (NO) and with the mechanisms of its action and synthesis in man and animals. The cytotoxic, vasodilatory, neuromediator, and other properties of NO are analyzed. NO is shown to perform many functions in health and in diseases of various genesis. Analyzing the structure and functions of NO in health and in diseases suggests that they all have both structural and functional properties and their own features. The currently available data lead to the conclusion that the NO-synthase mechanism that is responsible for the production of NO is its synthesis in the presence of oxygen. During functional exercises and hypoxia of various origin, another mechanism of NO may become active, which is associated with the reduction of NO2- in NO. PMID- 10832412 TI - [The biomedical aspects of nitric oxide cycles and of the superoxide anion radical]. AB - The paper presents a concept of the cycles of nitric oxide and superoxide radical anion and discusses the biomedical aspects of these cycles. The developed concepts and their related ideas suggest that the mechanism of the cycle(s) shows the naturally latent regularity and assigns a meaning to orderliness in the system of free radical compounds. By their chemical nature, the latter are well known to tend to behave aggressively and commonly unpredictably, which frequently causes abnormalities of various genesis. In the author's opinion, the mechanism of antiradical protection of cells and the body as a whole is built into the very cyclic arrangement of metabolic processes which are attended by the formation of free radicals. Failure of this cyclic mechanism may be a cause of many diseases. PMID- 10832413 TI - [Nitrogen monoxide and nociceptive processes]. AB - Experiments on conscious rats studied the effects of systemic or central application of a nitric oxide (NO) donor or NO-synthase inhibitor on the pain sensitivity threshold. Injection of SNAP in doses of 0.2, 2.0, and 20.0 micrograms into the subarachnoidal space of the ventral medulla through a preimplanted steel cannula was accompanied by a dose-dependent change in tail flick latency. Intraperitoneal injection of 30 mg L-NAME also increased pain sensitivity threshold levels. The findings suggest that the decrease in pain sensitivity after systemic NO-synthase inhibitor administration is due to the modulation of NO-dependent processes at both the central and peripheral levels. PMID- 10832414 TI - [The ultrastructural localization of NO-synthase NADPH diaphorase in a peripheral nerve and its change in diphtheritic polyneuropathy]. AB - Light and electron microscopy was used to study the distribution and changes of NADPH-diaphorase in the cutaneous nerve biopsy specimens in different periods of diphtheritic polyneuropathy (DP). there was a reduction in the reaction rate of the enzyme in Schwann's cells of the destructively changed nerve fibers and an increase in the remyelinated nerve fibers. The enzyme is located on the nuclear and endoplasmic reticulum membranes and ribosomes. It is suggested that there is an association of the synthesis of nitric oxide with the myelin-producing function of Schwann's cells. PMID- 10832416 TI - Health legislation in 2000. PMID- 10832415 TI - [Nitric oxide, hemoglobin and laser irradiation]. AB - The paper deals with the present views of some chemical properties and biological effects of nitric oxide (NO) and chiefly its formation and primary conversions. The interaction of NO with superoxide radical is shown to one of the most important reactions of the former, which gives rise to peroxynitrite whose breakdown yields a hydroxy radical. Emphasis is laid on the substances that are a temporary depot or a possible transport form of NO, such as nitrosothiols and nitrosyl complexes of non-hemic iron. NO is well-known to release when these compounds are degraded. It is suggested that hemoglobin is another NO depot, which forms stable complexes with the latter. These hemoglobin complexes may be degraded on exposure to laser radiation to form free NO that ha a vasodilatory effect. Photolysis of nitrosyl complexes of hemoglobin may be responsible for the therapeutical effect of laser radiation. PMID- 10832417 TI - How scared should we be of pesticides? PMID- 10832418 TI - An unusual pattern of bruising. PMID- 10832419 TI - A rebuttal to criticism of the Oregon Death with Dignity Act. PMID- 10832420 TI - Survivors of torture in a general medical setting: how often have patients been tortured, and how often is it missed? AB - OBJECTIVES: To measure the frequency of people reporting torture among patients in a medical outpatient clinic and to determine primary care physicians' awareness of their patients' exposure to torture. DESIGN: Cross-sectional survey followed by selected in-depth interviews of participants reporting a history of torture. Medical record review and interview of torture survivors' primary care physicians. SETTING: The internal medicine clinic of a large, urban medical center. PARTICIPANTS: A convenience sample of 121 adult patients who were not born in the United States and who were attending the adult ambulatory care clinic. INTERVENTIONS: All participants were interviewed using the Detection of Torture Survivors Survey, a validated instrument that asks about exposure to torture according to the World Medical Association definition of torture. Participants who reported a history of torture were interviewed in depth to confirm that they had been tortured. We reviewed the medical records of participants who reported a history of torture and interviewed their primary care physicians. MAIN OUTCOME MEASURES: Self-reported history of torture. The awareness of primary care physicians of this history. RESULTS: Eight of 121 participants (6.6% [95% confidence interval: 3.1%-13.1%]) reported a history of torture. None of the survivors of torture had been identified as such by their primary care physician. CONCLUSIONS: Physicians of patients who have not been born in the United States and who attend urban general medical clinics frequently are unaware that their patients are survivors of torture. Primary care physicians can be the locus of intervention in the care of torture survivors. The first step is for physicians to recognize the possibility of torture survivors among their patients. PMID- 10832421 TI - Health professionals cannot be silent witnesses. PMID- 10832422 TI - Epidemiology and control of enterobiasis in a developmental center. AB - OBJECTIVE: To determine if enterobiasis could be controlled in a developmental center. DESIGN: Population-based study. Annual screening of all residents by perianal swabs for enterobiasis and on admission or discharge. Treatment of infected residents and their contacts with mebendazole, 100 mg orally, with two doses given 14 days apart. MAIN OUTCOME MEASURES: The number of residents with enterobiasis and the cost of the program. RESULTS: The prevalence of enterobiasis fell rapidly and progressively, from 21% before mass medication to 1% after 3 years. CONCLUSION: Mass medication of residents with enterobiasis and their contacts was beneficial, harmless, and cost effective. PMID- 10832423 TI - The benefits of screening must outweigh the risks and costs. PMID- 10832424 TI - Excellent information is needed for excellent care, but so is good communication. PMID- 10832425 TI - Should we look further for cancer in a patient with venous thromboembolism? PMID- 10832426 TI - Simplifying physicians' response to domestic violence. PMID- 10832427 TI - Is globalization dangerous to our health? PMID- 10832428 TI - An introduction to cultural differences. PMID- 10832430 TI - Medical myth: Ten percent of patients who are allergic to penicillin will have serious reactions if exposed to cephalosporins. PMID- 10832429 TI - An innovative community medicine curriculum: the La Mesa housecleaning cooperative. PMID- 10832431 TI - How to manage head lice. PMID- 10832432 TI - [Clinical pathways in oncology]. AB - The diagnosis-related-group/prospective-payment system (DRG/PPS) was introduced into the health care system of the United States in 1983. This system triggered the development and implementation of clinical pathways aimed at reducing the length and cost of hospitalization. In Japan, trial use of a Japanese version of DRG/PPS was initiated in November 1998 in 10 hospitals under the control of the Ministry of Health and Welfare, and full-scale implementation of the system is expected in the near future. Clinical pathways, therefore, are a current focus of attention, mainly because of their success in enhancing management efficiency in the U.S. However, in actual clinical settings where clinical pathways are used, several Japanese health care providers have come to realize that they are also useful in improving staff coordination, patient satisfaction, and patient care, rather than simply reducing the length of hospital stay and cost of health care. The introduction of clinical pathways requires that treatment of the disease in question be defined and standardized. The implementation of pathways for the treatment of cancer, however, might prove difficult because of the high frequency of variation. In our experience, the main reason for the use of clinical pathways is not to reduce the number of variant cases but to provide high-quality care through the promotion of a team approach to treatment and enhanced patient care. Therefore, even if there were frequent variances following surgery for cancer, those occurring in accordance with the pathophysiological state of the patient would not interfere with management by clinical pathways. Clinical pathways are advantageous because they allow patients to know their treatment schedule; to prepare for hospitalization procedures; to have a better perspective on discharge; to reduce anxiety regarding hospital admission, even if it is the first time; to communicate better with doctors, nurses, and other medical care staff, leading to greater trust; and to improve their ability for self management. These features are all important for the improvement of patient care. Furthermore, clinical pathways may lead to a situation in which the cost of hospitalization can be predicted prior to admission, enabling patients to compare differences between several hospitals. From our experience with gastric cancer, breast cancer, and esophageal cancer management, we consider clinical pathways to be of great benefit in helping to reform the current medical care system in regard to the management of cancer patients as well as patients with other diseases. PMID- 10832433 TI - [Surrogate and true endpoints in cancer clinical trials]. AB - To facilitate the proper and quick evaluation of cancer chemotherapy, the issues of surrogate and true endpoints in clinical trials were reviewed. Surrogate endpoints are defined as response variables that can substitute for a true endpoint due to their close correlation with the true endpoint, and their occurrence earlier than true endpoint. The response rate in cancer chemotherapy has been used as a surrogate or true endpoint for the evaluation of treatments. However, recent studies revealed that response rates do not always correlate with survival time (time to failure), and better surrogate endpoints such as time to progression, median survival time, or various surrogate biomarkers are now under investigation. PMID- 10832434 TI - [Assessment of quality of life as an endpoint of cancer treatment]. AB - In order to evaluate the efficacy of cancer therapy, it is important to assess quality of life (QOL) as an endpoint in addition to tumor size reduction and survival. Thus, assessment of QOL has been included in many clinical trials of cancer therapy. These studies use various multiple QOL scales. We have translated the English version of the FACT-BL (Functional Assessment of Cancer Therapy for Bladder Cancer) questionnaire into Japanese. In the process, we performed a comparative study simultaneously using this questionnaire and EORTC QLQ-C30J which is generally used for various cancer patients. We found that FACT-BL provides a more sensitive assessment of the QOL of patients with advanced bladder cancer than EORTC QLQ-C30J. This means that we should carefully choose a questionnaire which is directly related to disease site, symptoms and treatment and can be conducted clinically. In the future, the assessment of QOL will have an important role as an endpoint of cancer treatment if the methodology and infrastructure, such as coordinators and data centers, are established in Japan. PMID- 10832435 TI - [Significance of prolonged NC as an endpoint of chemotherapy for solid tumors]. AB - The goals of chemotherapy in patients with cancer should be both tumor shrinkage and extension of survival time. Believing that there is a clear correlation between the extent of treatment-induced tumor reduction and survival time, we have made an effort to reduce tumor size. However, many investigators have found no positive correlation between response rate and median survival time in various cancers, including non-small cell lung cancer and gastric cancer. If this is the case, we should reconsider our therapeutic strategy. We previously reported that doctors should induce a prolonged dormant phase rather than strive to shrink tumor mass in "Tumor Dormancy Therapy," because the survival of most patients with solid tumors depends on the length of the induced dormant phase rather than on induced tumor reduction. In this paper, we analyzed two Japanese phase II studies of gastric cancer. There were no significant differences between the survival of NC patients with TTP of more than 90 days and CR + PR patients in either study. These results suggest prolonged NC could contribute to longer survival, and we concluded that NC patients with greater than 90 days of TTP, that is "prolonged NC," exhibited similar survival relative to those with effective treatments and tumor shrinkage, and should be evaluated as a positive response. PMID- 10832436 TI - [Correlation between response and survival]. AB - The true endpoints of cancer chemotherapy have to be prolongation of survival, palliation of symptoms, or improvement of quality of life in cancer patients, while the response rate is considered to be a surrogate endpoint. When conducting a phase III trial, not only the response rate but also data on survivals in the phase II study should be taken into consideration. New response evaluation criteria and various objective markers in analyzing survival have been developed recently. Our data on chemotherapy for gastrointestinal malignancies reveal correlations between CR cases and survival prolongation among esophageal cancer patients treated with chemoradiotherapy, and between responder and survival prolongation in gastric cancer patients treated with chemotherapy. A review of the literature might reveal a correlation between response and survival prolongation in colorectal chemotherapy patients, though our data did not support such a correlation. At present, it is necessary to recognize that "evidence" should be based on the results of randomized phase III trials with large sample sizes and good quality assurance other than the response rates in phase II trials. PMID- 10832437 TI - [Evaluation of therapeutic effect of chemotherapy with reference to clinical benefit response]. AB - In chemotherapy (CHT) for solid tumors, improvement of symptoms may be clinically beneficial, even if objective tumor responses are not recognized. The vast majority of patients with advanced pancreatic cancer (PC) are symptomatic at diagnosis. In particular, severe pain and poor general condition can be major debilitating problems. In patients with such symptoms, optimal palliation of symptoms to maximization the quality of life is of primary importance. In recent years, such a clinical benefit response (CBR) has become a new treatment end point being explored in clinical trials involving PC. The CBR is generally evaluated using two parameters in patients with PC: pain (intensity of pain and consumption of morphine) and Karnofsky performance status (KPS). A remarkable CBR was obtained in about 20-25% of patients treated with gemcitabine or combined CHT using fluorouracil and cisplatin (FP therapy). This new assessment method for CHT using CBR may be useful for evaluating chemotherapeutic response and may provide a strong rationale for continuing CHT in patients with stable disease according to imaging modalities. PMID- 10832438 TI - [Thymidylate synthase activity after preoperative administration of 5-FU in patients with gastric or colorectal cancer]. AB - Continuous intravenous injection of 5-FU was given at 300 mg/m2 to patients with gastric or colorectal cancer for consecutive 3 days preoperatively, and the relationships between the time until collection of samples (from final administration of 5-FU to excision of tissue samples) and total thymidylate synthase (TS total) activity, free thymidylate synthase (TS free) activity, thymidylate synthase inhibition rate (TSIR), thimidine kinase (TK) activity, and tissue 5-FU and FdUMP concentrations investigated. TS total was shown to gradually reduce with time, but the relationship between time and the other assay items could not be identified due to large variability in the data. TS total and TK also proved to be affected also by the sites at which the samples were collected, and exhibited significantly higher enzyme activity in tumor tissue than that in normal tissue. PMID- 10832440 TI - [Histoculture drug response assay on non-small cell lung cancer]. AB - We examined the chemosensitivity of non-small cell lung cancer (NSCLC) tissues to CDDP, 5-FU, ADM, MMC, ETP and SN38 using histoculture drug response assay (HDRA). One-hundred and thirty surgical specimens from NSCLC patients who were not given preoperative chemotherapy were used. The inhibition indexes of CDDP, 5-FU, MMC, ADM, ETP and SN38 were 39.1 +/- 18.2%, 48.0 +/- 19.7%, 63.3 +/- 17.7%, 47.6 +/- 22.0%, 36.9 +/- 21.1%, and 37.9 +/- 25.2%, respectively. Inhibition indexes were above the cutoff level, i.e., 'judged sensitive,' in 40 cases (31.3%) for CDDP, 34 cases (27.4%) for 5-FU, 54 cases (44.3%) for MMC, 36 cases (33.0%) for ADM, 29 cases (29.8%) for ETP, and 34 cases (37.4%) for SN38, respectively. In almost one third of patients, the inhibition indexes of all drugs were under cutoff levels. Correlations between in vitro chemosensitivity data and patient responses to chemotherapy were obtained from 16 evaluable patients, and a 44.4% true positive rate and a 100% true negative rate were observed. Our results with HDRA for NSCLC showed a high incidence of intrinsic multidrug resistance. HDRA may help doctors to avoid non-effective chemotherapy for NSCLC patients. PMID- 10832439 TI - [The effect of combination chemotherapy to adapted to chronotherapy with 5 fluorouracil, leucovorin, mitomycin C and cisplatin in patients with gastric or colorectal cancer]. AB - We performed combination chemotherapy adapted to chronotherapy with 5 fluorouracil, leucovorin, mitomycin C and cisplatin in 11 patients with gastric cancer and 7 with colorectal cancer. Treatment consisted of a 5-day course of continuous arterial or intravenous infusion of 5-FU (500 mg/body/day), arterial or intravenous infusion of leucovorin (20 mg/body/day) at 6:00 p.m. on days 1-5, arterial or intravenous infusion of mitomycin C (2 mg/body) at 9:00 a. m. on day 5, and arterial or intravenous infusion of cisplatin (20-80 mg/body) at 6:00 p.m. on day 5. The effective rate against gastric cancer was 73%; however, the effective rate against colorectal cancer was 29%. During and after this therapy, there was only a little appetite loss, nausea and stomatitis. PMID- 10832441 TI - [A pilot study of irinotecan hydrochloride for metastatic breast cancer--efficacy as a salvage therapy]. AB - From April 1996 through 1998, we treated twelve metastatic breast cancer patients with CPT-11 100 mg/m2 given on day 1, 8, and 15. All patients had received prior chemotherapy including anthracycline, and also another schedule such as MMC-VDS or HDMTX-5-FU. CPT-11 was well tolerated. One CR, two PR, and two long NC were achieved. Grade 3 leukocytopenia occurred in 2 patients. Grade 1 diarrhea occurred in 3 patients. According to the Kaplan-Meier method, the 50% survival was 354 days. In spite of intense prior chemotherapy, the treatment results with CPT-11 were satisfactory for anthracycline resistant metastatic breast cancer. PMID- 10832442 TI - [Effectiveness of carboplatin and etoposide selective intra-arterial chemotherapy for progressive soft-tissue sarcoma]. AB - Four patients with advanced soft tissue sarcomas were treated by selective intra arterial injection of carboplatin (450 mg/m2/2 hr) and etoposide (200 mg/m2/2 hr). After 2-4 courses of treatment, tumor size was remarkably reduced. Three of the 4 patients became candidates for limb-salvage surgery with a safety margin. During the treatment, patients showed no significant complications except for slight nausea and myelosuppression. This protocol might be a recommendable treatment for advanced soft tissue sarcomas. PMID- 10832443 TI - [A case of AFP-producing gastric cancer after curative operation effectively treated with chemotherapies including hepatic arterial infusion therapy]. AB - A 63-year-old woman was admitted complaining of epigastralgia. An endoscopic examination revealed a Borrmann type 2 lesion at the greater curvature of the middle third in the stomach. Abdominal computed tomography detected no liver metastasis. The preoperative serum alpha-phetoprotein (AFP) level was elevated to 242.9 ng/ml. 5-fluorouracil (5-FU) was given 200 mg/day for 26 days orally. Distal gastrectomy with D3+ No. 16b1 lymph node dissection, cholecystectomy and hepatic arterial canulation were performed, and mitomycin C 20 mg was injected intravenously during the operation. Immunohistochemical staining of the specimen for AFP by the SAB method was positive in the cancer lesion. After the operation, FP (cisplatin 100 mg on day 1, 5-FU 750 mg on days 1-3) therapy of one course and intrahepatic arterial infusion therapy using adriamycin 10-20 mg every 2 weeks for 7 months were conducted. Moreover, the patient took UFT 400 mg/day for 26 months orally on an outpatient basis as an adjuvant chemotherapy. The only toxicity was neutropenia (grade 3), but it abated without an interruption in the chemotherapy. The AFP level declined gradually, and returned to the normal range 1 month after surgery. The patient is still alive with no sign of hepatic metastasis or recurrence 7 years and 7 months after the gastrectomy. PMID- 10832444 TI - [A case study of alpha feto protein (AFP)-producing gastric carcinoma with multiple liver metastases, in which chemotherapy was effective enough to once save a life in rapid decline]. AB - A 57 year-old man visited our hospital with general fatigue and a sensation of abdominal fullness. He had lost 10 kg in body weight during the previous 3 months. Between admission and the time the diagnosis of AFP-producing gastric carcinoma with multiple liver metastases was made, his condition deteriorated quickly due to the rapid growth of the liver metastases. Combined chemotherapy consisting of cisplatin and 5-FU was given, and was so effective that the patient recovered well and both serum AFP level and the size of the swollen liver decreased markedly. However, about a month after being discharged, he experienced a relapse and was readmitted. After obtaining informed consent, chemotherapy consisting of methotrexate and 5-FU was started. Though the level of tumor markers and LDH decreased significantly, he died of hepatic failure. We think that this case is worthy of notice because it shows the effectiveness and limitations of chemotherapy in a situation where the condition of a patient is deteriorating quickly due to rapid extension of an AFP-producing gastric carcinoma. PMID- 10832445 TI - [Efficacy of UFT in the treatment of para-aortic lymph node metastasis following gastric cancer surgery: case report]. AB - The patient was a 68-year-old man who underwent pyloric gastrectomy for advanced stomach cancer on December 6, 1996. The histopathological diagnosis was poorly differentiated adenocarcinoma, ss, ly3, v1, n2 (+), and stage IIIa. Postoperative adjuvant chemotherapy consisted of short-term intravenous infusion of 5-FU, 320 mg/m2/day (= 480 mg/body) for 5 days beginning on postoperative day (POD) 1, and oral 5-FU, 200 mg/day, for 1 year beginning on POD 14. The preoperative CEA value was 316.2 ng, but it fluctuated below 10 ng postoperatively. About one year after the operation, the patient began to complain of epigastric pain, loss of appetite, and general malaise. CT of the upper abdomen revealed a 1.5-cm para aortic lymph node, and the CEA value of 319.0 ng was abnormally high. 5-FU was stopped, oral UFT at 300 mg/day was started, and the patient's course was followed. Three months after the start of UFT, the lymph node had shrunk on CT (shrinkage rate: 66.7%), and the CEA value had decreased to 14.3 ng. As though corresponding to these changes there was a gradual decrease in the epigastric pain, general malaise, etc., and the patient's appetite also returned. There were no subsequent elevations in the CEA values or increases in the size of the para aortic lymph nodes, and the patient's general condition was favorably maintained. UFT appeared to be effective against the lymph node metastasis around the aorta in this case. PMID- 10832446 TI - [A case of biloma caused by early complications of hepatic artery infusion chemotherapy]. AB - We report a 70-year-old male who had biloma as an early complication of hepatic artery infusion chemotherapy. The patient had a history of subtotal gastrectomy for a advanced gastric cancer. Two years after the primary operation on the stomach, a solitary metastatic liver tumor was indicated by follow-up abdominal CT, and a segmental hepatectomy was performed. Soon after the hepatectomy, intraarterial catheter placement was performed via the left subclavian artery for preventive chemotherapy. Infusion chemotherapy of 10.5 g 5-FU and 75 mg CDDP was administered for a month, during which time the patient had liver dysfunction, fever, tenderness, and abdominal fullness. Abdominal CT revealed a large low density mass at a lateral segment of the liver which could not be seen on the previous CT image. Also, extravasation of contrast media was identified by angiography via the reservoir catheter. Using an interventional technique, percutaneous transhepatic drainage for biloma and extubation of the reservoir catheter were performed. The present case is thought to be of an early and rare complication of hepatic artery infusion chemotherapy. The etiology is discussed herein. PMID- 10832447 TI - [A case of far advanced breast cancer with distant metastases which had mostly disappeared after chemoendocrine therapy]. AB - We report a case of far advanced breast cancer showing an excellent response to chemo-endocrine therapy. A 40-year-old female with a huge ulcerated tumor on her left anterior chest visited our hospital. Distant metastases were found in the lymph nodes, liver and bone. Therefore, endocrine therapy (toremifene and medroxyprogesterone acetate) and chemotherapy (cyclophosphamide, Therarubicin and 5-fluorouracil) were started as a combination treatment. As a result, the main tumor and metastatic lesion were remarkably reduced, and extended mastectomy with resection of right axillary lymph nodes was performed. Histologically, cancer cells in the primary lesion mostly disappeared, and only one lymph node in the left axillary lesion showed metastasis. No recurrence was found for 16 months after the surgical treatment. The combined therapy in the present case was extremely effective. PMID- 10832448 TI - [Efficacy of combined chemoendocrine therapy with doxifluridine, cyclophosphamide, and fadrozole hydrochloride hydrate in a case of stage T4N3M1 breast cancer]. AB - A 66-year-old female presented with a swollen lump in the left breast. She was diagnosed as having advanced breast cancer of stage T4N3 (supraclavicular lymph node) M1 (bone). The administration of CEF and TAM failed to improve her condition. After the treatment regimen was changed to combined chemoendocrine therapy with CPA, EPI, 5'-DFUR, and MPA, the areas of bone metastases were reduced. However, MPA caused side-effects (acute obstruction of the lower limb), and thus the treatment was discontinued after 4 months. Subsequently, the treatment combination was changed to CPA, EPI, 5'-DFUR, and fadrozole hydrochloride hydrate. After one year of the treatment, a complete response (CR) was obtained with the disappearance of the supraclavicular lymph node and bone metastases. After EPI reached the maximum administration amount, the remaining CPA, 5'-DFUR and fadrozole hydrochloride hydrate oral administrations were continued. As of 3 years and 10 months after the onset of the chemoendocrine therapy, CR has been maintained with suppression of the primary and metastatic lesions, without degrading the patient's quality of life. PMID- 10832449 TI - [Two cases of malignancy-associated hypercalcemia from bone metastases of breast cancer successfully treated with combination therapy using pamidronate and calcitonin]. AB - Two patients with malignancy-associated hypercalcemia from bone metastases of breast cancer, accompanied by consciousness disturbance, were treated by a combination therapy of pamidronate and salmon calcitonin. The cause of the hypercalcemia in both cases was thought to be expanded bone metastases, which induced a local osteolytic hypercalcemia (LOH). In the end, this regimen could not control the growth of the metastatic tumor, but it produced a more rapid and prolonged decrease in serum calcium level than a single agent, and resulted in lessened consciousness disturbance without adverse effects. Hypercalcemia is a life-threatening paraneoplastic syndrome which requires urgent medical treatment, since malignant hypercalcemia progresses very rapidly and induces several severe complications. In conclusion, this combination therapy was extremely effective for consciousness disturbance accompanying hypercalcemia due to widespread bone metastases of cancer. PMID- 10832450 TI - [Pharmacokinetics studies of nafamostat mesilate (FUT), a synthetic protease inhibitor, which has been used for the treatments of DIC and acute pancreatitis, and as an anticoagulant in extracorporeal circulation]. AB - Nafamostat mesilate (FUT) was first reported by Fujii et al, in 1981 as a synthetic protease inhibitor. FUT has been reported as a drug for the treatments of DIC (disseminated intravascular coagulation) and acute pancreatitis and as an anticoagulant in extracorporeal circulation. FUT has a structure of ester conjugate of p-guanidinobenzoic acid and 6-amidino-2-naphthol. In in vivo, this ester site was found as the reaction center as well as the site for the catabolic changes. Plasma half life (t1/2 beta) of FUT was about 23.1 min, compared to about 55 seconds of the related compound, FOY. The inhibitory activity of FUT on the protease was found to be due to the mis-reading of serine protease in vivo. PMID- 10832451 TI - [Phase I clinical trial design of anticancer agents--a Fibonacci and a modified Fibonacci sequence]. AB - A Phase I clinical trial of an anticancer agent is the first evaluation in humans, and it is an important step in drug development. From the ethical point of view, the goal is to escalate to the maximum tolerated dose quickly, yet safely, to minimize the likelihood of treating patients at doses that are too low or high. It is expected that the contradictions between safety and efficacy in the Phase I clinical trials will be solved by developing methods. The modified Fibonacci sequence has been generally adopted for dose escalation, although it includes some problems. It is necessary to recognize that the method used for Phase I clinical trials for anticancer agents remains unsatisfactory, and that it is also necessary to develop more ethical and scientific methods. PMID- 10832452 TI - Health profile of racial and ethnic minorities in the United States. PMID- 10832453 TI - The monitoring of racial/ethnic status in the USA: data quality issues. AB - This paper reviews the assessment of racial and ethnic identification in the major data collection systems of the US Department of Health and Human Services. It evaluates the quality of the available data and outlines recommendations for improving the collection of racial data and enhancing our understanding of the role of race in health. Special attention is also given to the role of socioeceonomic status in understanding racial differences in health and the assessment of racial status in data systems in the UK. PMID- 10832454 TI - Informal care and the empowerment of minority communities: comparisons between the USA and the UK. AB - This paper examines informal care and the empowerment of minority communities with respect to health care and health promotion in the USA and the UK based upon work prepared for the workshop, 'Involving Black and Minority Ethnic Users in Delivery of Services and Empowering Communities' presented during the bi-national 1997 USA, UK. Conference, 'Health Gain for Black and Minority Ethnic Communities' and the information gained from that Conference. 'Informal care' is operationally defined as 'the practice of alleviating distressful physiological and psychological dysfunctions through all others (e.g. traditional healers, family members, self, etc.) using measures that do not require a physician's prescription or intervention (e.g. lifestyle modifications) typically outside of formal, institutionally based care mechanisms (e.g. homes and communities). Informal care is a significant force in health maintenance, health promotion, and disease prevention. In the USA, at least one-third of the population is estimated to engage in unconventional medical practices, and perhaps, one-half of racial/ethnic populations use informal care. An enormous potential exists to better utilize informal care because informal care is culturally more compatible, relatively low cost, and flexible. The policy of the Indian Health Service in accepting the use of traditional medicine was cited. The US Congress recognized the potential of alternative medicine by establishing such an Office within the National Institutes of Health. 'Empowerment of racial/ethnic minority communities is the right for minority populations to determine their own destinies. In the USA, racial/ethnic minority populations are Blacks, Hispanics, Asians or Pacific Islanders, and American Indian/Alaska Native. These classifications are based upon self-report; in the UK, the black and minority classifications are based upon countries of birth rather than self-reported racial/ethnic identities. Empowerment of these communities is important both demographically and historically. In demographic terms, racial/ethnic minority populations are increasing at higher rates than the majority population in the USA and hence, the health status of minorities will become the health status of the nation in the next half century. Historically, racial/ethnic minorities have not been empowered. As a consequence of the 1985 Secretary's Task Force Report on Black and Minority Health Federal measures to address disparities in the health status of minorities were initiated. In March 1994, the UK Government initiated the Ethnic Health Unit within the National Health Service. These measures are not mature enough to evaluate their impact. However, progress in implementing measures to empower minorities in the UK have begun and are illustrated by the work reported by Dr Pui-Ling Li, the UK counterpart to the workshop, 'Involving Black and Minority Ethnic Users in Delivery of Services and Empowering Communities'. Recommendations are made to increase use of informal care and the empowerment for racial/ethnic minority communities and to build upon the works in progress in both the USA and the UK. PMID- 10832455 TI - Quality of health care for ethnic/racial minority populations. AB - This article reviews the composition and characteristics of the health professions, the demographics of the national population, and factors that influence access to health care and satisfaction with care for ethnic/racial minority populations in the United States. In addition, an overview of publicly funded US health insurance programs for the poor is provided along with a discussion of the impact that managed care is having on the American health care system. Finally, the paper summarizes conference discussions regarding the problems, strategies, and approaches that the UK and the US have experienced with respect to providing quality health care for ethnic/racial minority populations. PMID- 10832456 TI - Primary care: choices and opportunities for racial/ethnic minority populations in the USA and UK--a comparative analysis. AB - This paper examines and compares the choices made and the opportunities provided by the United States and the United Kingdom in delivering primary care services to their racial/ethnic minority populations. While both nations agree that the most effective strategy for health service delivery to a diverse population lies in primary care, their approaches to obtaining this goal have been quite different. Sociological theories of functionalism and conflict perspective provide the analytical and organizing framework of the paper. Within this theoretical context, the health systems in place in each country are examined as an outgrowth of the larger socio-political, economic and cultural structures of the US and UK. Analysis of the advance of managed care in the US and the recent NHS reforms are also discussed in terms of lessons learned and the difficulties that lay ahead in order to ensure that these new developments contribute significantly to eliminating the disproportionately worse health status of racial ethnic minorities. Towards that goal the paper identifies opportunities for collaboration and specific recommendations for future action by both countries. PMID- 10832457 TI - Obstacles to equality: issues for purchaser and provider. PMID- 10832458 TI - The progress of Indian states and the new strategy for child and family welfare. PMID- 10832459 TI - Controversies in pediatrics. PMID- 10832460 TI - A to Z: vitamin A and zinc, the miracle duo. AB - Dietary micronutrients such as vitamins and trace minerals are known modulators of host immune responses against common pathogens. In this respect, vitamin A and zinc have recently received increased attention. Several in vivo and in vitro studies suggest that vitamin A may be a critical player in the mucosal immune responses in the respiratory and gastrointestinal tracts, particularly in undernourished children. The effect may be mediated primarily by stabilization of the membrane of mucosal epithelial cells, as well as enhanced leukocyte functions. The beneficial effect of vitamin A therapy in reducing measles associated morbidity and mortality suggests its crucial role in defenses against viral pathogens. Zinc is also known affect leukocyte functions such as phagocytosis and T-lymphocyte-mediated immune responses. However, unlike vitamin A, zinc has been investigated primarily for its effects on bacterial infections. Dietary supplementation or therapeutic treatment with vitamin A and zinc may be a cheap yet effective means of preventing or treating infections in highly susceptible populations. Additional studies, however, are required to better define the types of pathogens and the specific human populations that may benefit from such therapy. PMID- 10832461 TI - Do we need to treat otitis media? AB - Otitis media is a common pediatric problem. It is well established that over half of infants and children with acute otitis media may have spontaneous recovery. Since it is difficult to predict the course (self-limited versus serious disease) all the children with acute suppurative otitis media need to be treated with antibiotics. Amoxicillin is still the initial antibiotic of choice. There are several alternate antibiotics available with activity against beta-lactamase positive bacteria. These agents have no advantage over amoxicillin in infections due to penicillin resistant pneumococci. Recent use of beta-lactam antibiotics and/or attendance in a day care where there is frequent use of antibiotics are predisposing factors for penicillin resistant pneumococcal infection. In such cases after tympanocentesis, higher dose of amoxicillin, clindamycin or intramuscular ceftriaxone should be considered. Secretory otitis media does not need to be treated with antibiotics unless the patient is in high risk group. Prophylactic use of antibiotics should be actively discouraged. Influenza and pneumococcal vaccination (2 years or older) should be encouraged in children with recurrent episodes of acute otitis media. Breast feeding should be encouraged. PMID- 10832462 TI - Streptococcal pharyngitis: is penicillin still the drug of choice? AB - With the advent of cephalosporins, penicillin appears to have lost some ground for treatment of Acute Group A Beta Hemolytic Streptococcal Pharyngitis. It has been argued for some time now whether penicillin should remain the drug of choice for the management of this infection. Evidence has been presented both in favour and against using penicillin for Group A beta hemolytic streptococcal (GABHS) pharyngotonsillitis. In this commentary, we review the available data in the current literature and conclude that penicillin should still remain the drug of first consideration for GABHS pharyngitis. If penicillin treatments were less effective now, the clinical and bacteriologic failure rates should be on the rise compared to the previous years. PMID- 10832463 TI - Gastroesophageal reflux--how to mend it? AB - Gastroesophageal reflux (GER) is a common condition affecting children. The clinical presentation varies widely from innocuous spitting up to life threatening apnea. Various diagnostic tests are available to document the etiology, presence or complications of GER. In most cases, conservative approach is sufficient. In complicated cases, pharmacotherapy is indicated while surgical therapy is reserved for resistant cases with complications. PMID- 10832464 TI - Cytokines in infectious diseases. AB - Cytokines are heterogeneous group of proteins that are produced by a wide variety of cells in the body and act as signals between the cells of the immune system. The recent discovery of two distinct subsets of T-helper cells led to the development of a conceptual framework of complex network of cytokine pathways in the regulation of immune responses against a variety of infectious agents. In this brief review, we have attempted to present a concise summary of the current state of knowledge of this complex interaction among cytokines that result in a cascade of biological events leading to inflammatory response and host defense against pathogens. PMID- 10832465 TI - Brachytherapy for solid tumors in children. AB - The standard treatment after surgery and chemotherapy in pediatric solid tumors is external beam to the tumor with a generous (5 cm) margin for local control. This treatment is given over a five to six-week period, requires use of repeated deep sedation, and leads to unacceptable morbidity (especially organ and bone growth retardation) in infants and younger children. Limited volume irradiation by brachytherapy over a few days may be sufficient therapy for children treated with aggressive chemotherapy. Brachytherapy allows high doses of radiation to be limited accurately to the tumor bed, spares the surrounding normal tissues, and thus minimizes late sequelae. Manually afterloaded removable iridium-192, iodine 125, and cesium-137 have been used with good results. The major disadvantages of LDR brachytherapy are: the necessary sedation and immobilization of younger children to prevent accidental removal of the implants during the entire period, radiation exposure to the medical personnel and the parents, and the psychological effect of separating parents from their child. Using a low energy radionuclide such as iodine-125, or remote afterloading technology with LDR and PDR reduces radiation exposure hazards, but prolonged sedation and immobilization are still required. HDR brachytherapy not only eliminates the radiation exposure hazards but, in addition, eliminates the other disadvantages of brachytherapy thereby extending treatment to the infants and younger children. The long term effects of brachytherapy need further study. PMID- 10832466 TI - Chronic diarrhea: causes, presentation, and management. AB - Important inroads are being made into understanding the pathophysiology of diarrhea. Clear understanding of key mechanisms should suggest new approaches to combat disease. Exciting developments are occurring in terms of super-ORS solutions, particularly with the promise of short chained glucose polymers and glutamine. Perhaps the most important development is the prospect of a good rotavirus vaccine being available before the end of the decade. Chronic diarrhea seems to be on the increase globally, probably because of the success of ORS. The mechanisms that lead to mucosal injury are elusive, and therapy still largely supportive and empiric. Celiac disease continues to be a puzzle, because of the uncomfortable feeling that a majority of cases may be missed because of atypical presentations. The successful use of long term parenteral nutrition has allowed survival and better characterization of cases that otherwise would have perished as 'lethal protracted diarrhea'. Microvillus inclusion disease may be the commonest congenital secretory diarrhea. The role of the recently reported high prevalence of glucoamlase deficiency may be important. Lastly, attention to micronutrients, particularly low vitamin A and probably zinc may prove to be important in prevention and amelioration of diarrhea and growth failure. PMID- 10832467 TI - Reduced-osmolarity oral rehydration salts solution multicentre trial: implications for national policy. AB - World Health Organization (WHO) recommended standard ORS solution has sodium (90 mmol/L) and glucose (111 mmol/L) almost in the ratio of 1:1 and a total osmolarity of 311 mmol/L. There are concerns that the sodium or glucose concentration and the overall osmolarity in the formulation is not appropriate. Therefore, the efficacy of standard and reduced-osmolarity ORS solutions in young children with acute diarrhea was evaluated in a recent WHO supervised multicentre trial conducted in India (New Delhi), Brazil, Mexico and Peru. The implications of trial results are discussed. In non-cholera diarrhea, both the standard and reduced osmolarity ORS solutions were effective in achieving clinical rehydration. The stool output was 39% higher in the standard ORS solution group as compared to the reduced-osmolarity ORS solution group. The duration of diarrhea was 22% higher in the standard ORS solution group. The risk of requiring supplementary intravenous infusion was increased in children treated with standard ORS solution [relative risk 1.4 (0.9-2.4)]; this benefit was not observed in Indian patients due to high breast feeding rate. The mean sodium concentration at 24 hours after admission was lower in the reduced osmolarity ORS solution group [135 (134-136) vs 138 (136-139), p < 0.01). The low osmolarity ORS deserves to be evaluated in adult cholera to determine its efficacy and any excess hyponatreamia. Meanwhile, it is reassuring that the WHO formulation was effective and its use was not associated with hypenatremia even in young children. Efforts must continue to be made to promote WHO-ORS while research to improve it further is welcome. PMID- 10832468 TI - Polio eradication--target 2000. AB - Poliovirus an enterovirus is of 3 types (1, 2 & 3). The 1 poliovirus is most often the cause of paralysis. Poliomyelitis can be eradicated from human soil as man is the only reservoir of this infection and effective vaccines are also available for its control. Inactivated poliovirus vaccine (IPV) confers mainly the humoral immunity in comparison to oral polio vaccine (OPV) which gives the intestinal immunity as well. OPV has been recommended by the WHO as the vaccine of choice for global eradication of polio because of its superior ability to inhibit spread of wild polio virus, low cost and its ease of administration. 70 90% of polio cases occur in children less than 3 year of age. Each paralytic case which is the tip of an iceberg probably represents 100 to 1,000 infected persons in the community. The incidence of poliomyelitis is on the decline with 145 of 213 countries today report 0 case of polio. Central Africa & South Asia are the principal reservoirs of wild poliovirus with nearly two thirds of cases being reported from Indian subcontinent. Components of eradication strategy are: sustained high levels of immunisation, annual mass vaccination campaigns of OPV to all children under 5 years of age, establishment of extremely sensitive surveillance systems and targeted immunisation to areas and populations where poliovirus transmission is likely to persist. The task of global eradication of poliomyelitis is uphill but well within our reach. A strong will and political commitment by the Government of India is leading the nation to the goal of polio free world by the year 2,000. PMID- 10832470 TI - Juvenile rheumatoid arthritis--Madras experience. AB - In a prospective study of 1,053 consecutive children who attended the Rheumatic Care Centre, Government General Hospital, Madras from 1991 to 1995, 331 children fulfilled the criteria proposed by the American Rheumatism Association as modified by Cassidy et al for the diagnosis of Juvenile Rheumatoid Arthritis. These children were thoroughly examined and investigated and classified into 3 onset types which was then sub-classified into early entry and late entry groups based on the duration of illness. Other arthritic conditions were excluded. There were 44 cases belonging to Systemic onset, 171 belonging to polyarticular onset and 116 belonging to oligoarticular onset type. In the systemic onset type 44/44 patients had fever, 40/44 had lymphadenopathy and 19/44 had skin rash; wrists and knees 31/44 were the most commonly involved joints; neck involvement was present in 13/44 of the cases; ANA was positive in 5/44 cases and anaemia was seen in 24/44 cases. In polyarticular onset type wrists 119/171, knees 143/171, hip joints 105/171 and ankles 113/171 were commonly involved; in the RF +ve subtype 3/23 had subcutaneous nodules and 7/23 were positive for ANA; in the Rf -ve subtype 59/148 were positive for ANA. In the oligoarticular subtype-1 ANA was positive in all cases but iridocyclitis was not seen in any case. In oligoarticular subtype-2 HLA B27 was positive in 13/26 cases while Sacroilitis was seen in 16/26 cases. In oligoarticular type-3 HLA B27 was negative. PMID- 10832469 TI - Systemic lupus erythematosus. AB - Systemic Lupus Erythematosus (SLE) of childhood is a complex and challenging disease which can occur at any age. Identification of disease early in it's course and aggressive, appropriate management leads to improved outcome for an individual child. The history of SLE indicates how much progress has been made in the last quarter century. A discussion of the etiopathogenesis of SLE demonstrates the complexity of the syndrome. This is followed by a description of clinical manifestations, including diagnostic criteria, differential diagnosis and suggested methods for eliciting important symptoms to make the diagnosis. Evaluation of specific organs is next reviewed highlighting critical organ manifestations that are significant for future prognosis. Treatment of SLE includes a variety of medications, including non-steroidal anti-inflammatory medications, steroids and immuno-suppressive drugs. Attention to physical activity, stress and nutrition is equally important. Signs and symptoms that indicate disease flare or infection are described. Lastly, related syndromes are reviewed. PMID- 10832471 TI - Perinatal autopsy--a seven-year study. AB - A retrospective analysis of autopsies conducted on perinatal deaths during 7 years period (Oct 1983 to Sept 1990) was done. There were 650 neonatal deaths and 944 still births during the study period. Autopsy rates among neonates and still births were 33% and 4.9% respectively. There ware significant findings in 97.2% of neonatal deaths and in all still births. Pulmonary lesions followed by congenital malformations were the major pathological findings. Infection was observed among smaller number of babies compared to other Indian studies. Autopsy revealed many internal congenital malformations which were not diagnosed clinically. Autopsy changed or added to clinical diagnosis in 59.5% of cases. Perinatal autopsy is highly productive in our set up. PMID- 10832472 TI - Intravenous immunoglobulin for prophylaxis of nosocomial sepsis. AB - A total of 76 premature newborn infants with gestational age of 34 weeks or less were enrolled in a randomized controlled study to determine whether intravenously administrated immunoglobulin (IVIG) is able to prevent nosocomial sepsis. Forty infants were given 0.5 g/kg IVIG on the first day of life and 36 infants with similar gestational age and birth weight were selected as controls and did not receive IVIG. The frequency of proven sepsis, with a positive blood and/or cerebrospinal fluid culture, was significantly lower in infants who received IVIG as compared to controls (42.5 vs 80.0%) (p < 0.01). The mortality rate attributable to infection was not different in IVIG recipients and in controls (41 vs 48%) (p > 0.05). The overall mortality rates in the two groups were not different either (35.0 vs 44.4%) (p > 0.05). The majority of micro-organisms isolated from the blood culture of the patients were gram negative microorganisms (Klebsiella, Enterobacter). IVIG therapy was believed to be effective for prophylaxis of nosocomial infection, but such therapy was not able to reduce overall mortality rate or mortality rate due to systemic infection in prematurely born infants in our intensive care unit where the causative pathogens are usually gram negative microorganisms. PMID- 10832473 TI - The offspring of epileptic mother. AB - The offspring of an epileptic mother is an issue-currently getting attention because of its several implications. A complex interaction between epilepsy during pregnancy and its adverse impact on foetus, labor, neonate, congenital malformation, psychosocial and medico-social concern and treatment challenges of such cases is increasingly being realised. Some of the significant observations has been reviewed extensively in this article. Maternal epilepsy is likely to adversely affect the off-spring at its various stages of development amounting to increased morbidity and mortality. Increased seizure frequency during pregnancy with resultant increased risk is well documented but its mechanism is poorly understood. Low apgar score, increased still birth rates (1.3 to 14%) in offspring of epileptic mother (OEM) is reported. So also, the neonatal and perinatal deaths are twice more common in OEMS than normal control. Small for dates, and prematurity in OEM is reported to be 7 to 10% and 4-11% respectively. Adverse impact on labor and delivery like preclampsia, abruptio placentae, polyhydramnios, assisted delivery, cesarean section and IUGR poses particular challenges to the obstetrician. Pediatrician's alertness is needed to anticipate and deal with the bleeding manifestation due to deficiency of Vit-K dependent clotting factors and various anticonvulsant drug (AED) withdrawal symptoms. Significant risk of developing congenital malformation is the result of epilepsy perse and the AED used during pregnancy. AED exposure leads to other distinct clinical syndromes, the orofacial clefts and cardiac anomalies being the commonest manifestation. Epilepsy in mother but not in father has significant adverse impact. Management strategies in the context of available observation has been discussed. PMID- 10832474 TI - Fragile X syndrome among children with mental retardation. AB - Prospective screening for fragile X syndrome was carried out among 1,111 patients with mental retardation who attended the Genetic clinic. Using defined clinical criteria, 55 patients were selected for cytogenetic studies to detect folate sensitive fragile sites. Twenty patients were diagnosed to have the fragile X syndrome. The prevalence of fragile X (A) syndrome was 18 per 1,000 patients of both sexes with mental retardation, 2.8% among male patients with mental retardation, and 5.8% among subjects with nonspecific mental retardation. PMID- 10832475 TI - Cytomorphology of some uncommon tumors of childhood and adolescence. AB - With the advent of newer adjuvant chemotherapeutic regimes resulting in dramatic improvement in survival, it is mandatory to obtain quick correct diagnosis, which is provided by fine needle aspiration cytology (FNAC). Cytology of childhood tumors was studied to find any distinguishing features, which would help in arriving at a diagnosis. Cytomorphology of 386 childhood and adolescent (0-19 years) tumors diagnosed between 1984 and 1993 were studied. Lipoma was the commonest benign tumor in 0-14 years age group. Fibroadenoma of the breast was the common benign tumor in adolescent (10-19 yrs) girls and pleomorphic adenoma in adolescent (15-19 yrs) boys. Wilms' tumor was the most frequent solid malignant tumor in preschool children and Hodgkin lymphoma, mixed cellularity in the school going age (5-19 yrs). Malignant tumors were more frequent (199). In addition, some rare malignancies like hepatoblastoma, meningioma, ganglioneuroblastoma, adrenocortical carcinoma, clear cell sarcoma kidney and malignant histiocytosis were diagnosed. The cytological features and the differential diagnoses are discussed. It is possible to diagnose these uncommon tumors if particular attention is paid to the distinguishing cytomorphological features in correlation with clinicoradiological findings and cytochemistry. PMID- 10832476 TI - Sino-bronchial syndrome in children with asthma. AB - Thirty children in the age group of 2 to 12 years were brought with a history of recurrent non-seasonal moderate to severe wheezy episodes associated with symptoms of nasal congestion, sneezing and occasional headache. All of them had maxillary or pan sinusitis with 26 having associated right, left or bilateral lower lobe pneumonitis or bronchiectasis. Serum immunoglobulins were normal in 22 and was not done in eight. There was positive (2 to 4+ above negative control) skin test response to dust and dust mite in 15 of the 22 children tested. Throat swabs/sputum or nasal secretions grew B-hemolytic streptococcus or streptococcus pneumoniae in twenty-seven. All the children were put on bactericidal drugs for 6 to 8 weeks and bronchodilators were used when needed. At the end of 6 to 8 weeks follow-up X-ray of sinuses and chest showed significant clearing of the lesions which coincided with marked clinical improvement. Sinus X-ray should be considered in bronchial asthma resistant to medical management since untreated bacterial sinusitis can be an underlying cause of chronic poorly controlled asthma. PMID- 10832477 TI - Sonographic renal length in Indian children. AB - For the purpose of making norms of ultrasonographic renal length in Indian children, 240 children (137 boys and 103 girls) below 5 years of age were subjected to sonographic evaluation in supine position. Left renal length increased from 59.18 mm, in infancy to 81.89 mm between 4-5 years age in female children. Similar increase was observed in male children. There was no statistically significant difference in renal length in boys and girls in various age groups (p 0.05 to 0.1). Right renal length measured 1-3.5 mm than less than the left renal length, and this difference was statistically significant (p 0.05 to 0.0002) in both sexes. Renal length positively correlated to subject's age, length/height, body weight and body surface area in that order. PMID- 10832478 TI - Bilateral basal ganglia lucencies following acute febrile illness. AB - Bilateral striatal necrosis in children without damage elsewhere in the brain can present as an acute neurological disorder or as a progressive disorder. Three children of 6, 7 and 12 years age developed dystonic posture of limbs without any cranial nerve involvement or alteration of sensorium soon after recovery from acute high grade febrile illness of 3-4 days duration. Computerized tomographic scan of head showed bilateral necrosis of basal ganglia. We think that these patients probably constitute a clinically and radiologically distinct subgroup of disorder that produce bilateral striatal necrosis in children. The cause of the syndrome is unknown. PMID- 10832479 TI - Congenital segmental dilatation of intestine. AB - We are reporting two cases of segmental dilatation of small intestine presenting as neonatal intestinal obstruction. Clinco-pathological features, etiology and management are discussed in brief. PMID- 10832480 TI - Cecal duplications: a rare cause for secondary intussusception. AB - Duplications of the alimentary tract are rare congenital anomalies that may occur at any level from mouth to anus. While the oesophagus and the ileum are the most common sites, duplications of the colon are rare. Two cases of ileocolic intussusceptions in 8-month-old girl and 6-month-old boy who were admitted to our hospital with acute abdomen findings are presented. Intraoperatively, cecal cystic duplications leading intussusception were revealed. Intussusception is one of the most important surgical emergence in infancy and typically, it does not involve a lead point in childhood. Although duplication cyst may act as lead point, the review of literature reveals its rarity. PMID- 10832481 TI - Congenital megalourethra. AB - Megalourethra, a rare congenital disorder involving the anterior urethra, is subdivided into two types: Fusiform and scaphoid. Two cases of scaphoid type megalourethra are reported. The first patient, a 5-weeks-old infant diagnosed at birth as having the prune belly syndrome was admitted to the hospital with vomiting and failure to thrive. Intravenous pyelogram revealed marked dilatation of the left ureter and a bladder diverticulum. A penile urethrostomy was performed. The second patient, born to healthy parents after an uneventful pregnancy and delivery, was found to have an enlarged and deformed penis. The baby voided with a poor stream and a concomitant swelling of the penis was noted. Retrograde uretrography showed a sac-like dilatation of the penile urethra. Surgical revision was carried out a 2 stage procedure and was completed (Nesbitt's operation) after 4 months. The patient did well postoperatively and voided with a normal stream without any abnormality of the penile shaft. PMID- 10832482 TI - Septic arthritis due to Actinobacillus actinomycetemcomitans. AB - Actinobacillus actinomycetemcomitans, a gram-negative aerobic bacillus of the family Parvobacteriaceae which is a normal inhabitant of the oral flora, is a rare cause of human infection. We report a case of septic arthritis caused by this organism in an uncompromised child. PMID- 10832483 TI - Congenital deficiency of factor VII. AB - A case of congenital factor VII deficiency in a five-year-old child is reported. The patient, born of a non-consanguineous marriage, presented with repeated bouts of epistaxis since childhood. The prothrombin time (PT) was markedly prolonged with a normal bleeding time (BT), partial thromboplastin time with Kaolin (PTTK) and platelet count. The patient has been on follow up for the last four years and is doing apparently well. PMID- 10832484 TI - Hypomelanosis of Ito. AB - A two-year-old male child presented with bizarre hypopigmented skin lesions, severe mental retardation and generalized tonic-clonic seizures. Examination showed hypopigmented patterned whorls and irregular patches over the trunk and linear streaks over the flexor aspects of upper and lower limbs. He also had generalized hypertonia and brisk tendon reflexes. Other systems were normal. PMID- 10832485 TI - Prevalence of antibodies to West Nile fever, sandfly fever Sicilian, and sandfly fever Naples viruses in healthy adults in Israel. AB - BACKGROUND: The arbovirus infections West Nile fever (WNF) and sandfly fever (SFF) are both endemic in the Middle East. Despite the fact that the mosquito and sandfly vectors of these viruses are still found in Israel, SFF has not been reported since 1948, whereas outbreaks of WNF occur periodically. It is suspected that some of the cases of nonspecific febrile illnesses in Israel may in fact be either WNF or SFF. AIM: To determine the extent of past arbovirus infections in subgroups of Israeli soldiers. METHODS: Sera from random samples of three groups of healthy soldiers aged 18-20 (n=273), 21-30 (n=497), and 40-55 years (n=285) were examined for IgG and IgM antibodies to West Nile virus (WNV), sandfly fever Sicilian virus (SFSV), and sandfly fever Naples virus (SFNV), using an ELISA method. RESULTS: The prevalence of antibodies increased with age for all three viruses examined, and ranged from 7.0%, 0, and 2.8%, for WNV, SFSV, AND SFNV, respectively, in soldiers aged 18-20 years, to 41.9%, 23.7%, and 30.8%, in those aged 40-55 years. The percentage of seropositives for IgG who were also IgM positive were 1.0%, 5.9%, and 3.4%, for WNV, SFSV, AND SFNV, respectively. 17.5% of the seropositives for WNV were also positive for SFSV, as compared with 5.7% among the WNV-seronegatives. Among the seropositives for WNV, 23.99% were positive for SFNV, compared with 9.1% of WNV seronegatives. 48.2% of seropositives for SFSV were positive for SFNV, as compared with 8.4% for SFSV seronegatives. In the age agroup 40-55 years, there were significantly higher prevalence rates in those born in Israel compared with those born abroad. CONCLUSIONS: There is still significant exposure to WNV, SFSV, and SFNV in Israel. Arborviruses are probably responsible for many cases of non-specific febrile illness in this region. PMID- 10832486 TI - Measles control in Israel: a decade of the two-dose policy. AB - BACKGROUND AND METHODS: The introduction of routine measles immunization in Israel in 1967 was followed by a 95% reduction in reported measles incidence. In 1990, a second measles immunization dose was instituted, and up until the end of 1999, 16 birth cohorts were offered the second dose. We present here changes in reported measles incidence in Israel following institution of the two-dose policy. RESULTS: First-dose coverage is 94%, and coverage for the second school based dose exceeds 95%. A further 90% reduction in measles incidence has been observed and, following a modest national outbreak in 1994, measles incidence for 1995-99 stands at less than 3/100,000/year. CONCLUSIONS: Since measles is highly contagious, very high immunization coverage rates will be required to preserve these accomplishments, and measles elimination is still years away, but there is tentative evidence that measles containment is at hand. Even greater measles control can be anticipated as vaccine immunogenicity improves and successive cohorts of children come under the 2-dose regimen. PMID- 10832488 TI - Evidence of antioxidant role in disease prevention lacking, says panel. PMID- 10832487 TI - FDA backs away from elements of plan to regulate ephedrine supplements. PMID- 10832489 TI - Government study details effects of insurance coverage on consumer drug costs, use. PMID- 10832490 TI - Report identifies barriers to providing health care on the Internet. PMID- 10832491 TI - CliniTrend gets more horsepower. PMID- 10832492 TI - Seeking patient information outside your health system. PMID- 10832493 TI - Ginger. PMID- 10832494 TI - Uncertain quality of dietary supplements: history repeated. PMID- 10832495 TI - Impact of Parkinson's disease and its pharmacologic treatment on quality of life and economic outcomes. AB - The impact of Parkinson's disease (PD) and its pharmacologic treatment on health related quality of life (HRQL) and economic outcomes is reviewed. PD is a chronic and progressive neurologic disorder characterized by specific motor deficits resulting from the degeneration of dopaminergic neurons in the substantia nigra. The cardinal symptoms are tremor, rigidity, bradykinesia, and loss of postural reflexes. PD markedly reduces HRQL and places an economic burden on society of up to $25 billion per year. Patients' inability to move freely and to perform everyday tasks restricts their independence and leads to increased reliance on caregivers and assistive devices. Emotional and psychosocial well-being is also negatively affected. As the disease progresses, the response to levodopa typically decreases and various motor complications develop; these are difficult to treat and result in further declines in HRQL. The economic costs of PD include both direct health care costs (for drugs, physician services, and hospitalization) and indirect costs (for lost worker productivity). Economic analyses of PD and its treatments can help guide effective allocation of health care resources. Various antiparkinsonian agents and formulations, such as extended-release levodopa-carbidopa and pramipexole, have been found to be cost effective relative to other agents. The newest antiparkinsonian drugs, cathechol O-methyltransferase inhibitors, also have the potential to improve HRQL and economic outcomes, although more study is needed to confirm this. The total impact of PD and its treatment can be fully appreciated only when HRQL and economic outcomes, in addition to clinical outcomes, are examined. PMID- 10832496 TI - Content versus label claims in ephedra-containing dietary supplements. AB - The content of ephedra alkaloids in herbal dietary supplements containing ephedra (ma huang) was studied. The ephedra alkaloid content of 20 ephedra-containing supplements was determined by high-performance liquid chromatography. Contents of (-)-ephedrine, (+)-pseudoephedrine, (-)-methylephedrine, (-)-norephedrine, and (+)-norpseudoephedrine were measured. Ephedra alkaloid content varied considerably among products. Total alkaloid content ranged from 0.0 to 18.5 mg per dosage unit. Ranges for (-)-ephedrine and (+)-pseudoephedrine were 1.1-15.3 mg and 0.2-9.5 mg, respectively. (+)-Norpseudoephedrine, a Schedule IV controlled substance, was often present. Significant lot-to-lot variations in alkaloid content were observed for four products. For one product, lot-to-lot variations in the content of (-)-ephedrine, (+)-pseudoephedrine, and (-)-methylephedrine exceeded 180%, 250%, and 1000%, respectively. Half of the products exhibited discrepancies between the label claim for ephedra alkaloid content and actual alkaloid content in excess of 20%. One product was devoid of ephedra alkaloids. Assay of 20 ephedra-containing dietary supplements showed that alkaloid content often differed markedly from label claims and was inconsistent between two lots of some products. PMID- 10832497 TI - Changes in pharmacy, nursing, and total personnel staffing in U.S. hospitals, 1989-1998. AB - Pharmacy, nursing, and total hospital personnel inpatient staffing in U.S. medical-surgical hospitals for 1989, 1992, 1995, and 1998 was studied. Nursing and total personnel staffing data were obtained from the American Hospital Association, and pharmacy personnel data were obtained from the National Clinical Pharmacy Services database. Between 1989 and 1998, mean +/- S.D. registered-nurse staffing per 100 occupied beds increased from 124.46 +/- 92.24 to 196.57 +/- 131.92, or 57.94%. Pharmacist staffing per 100 occupied beds increased from 6.47 +/- 3.01 to 7.95 +/- 4.88, or 22.87%. The total number of registered nurses increased by 126,960 (15.78%), and the total number of pharmacists declined by 320 (0.72%). The increase in pharmacist staffing per 100 occupied beds was due almost entirely to decreases in hospital census between 1989 and 1998. Total hospital personnel staffing per 100 occupied beds and the absolute number of hospital employees increased at much higher rates (55.2% and 12.95%, respectively) than pharmacist staffing. Pharmacy technician staffing per 100 occupied beds increased by 42.96%, and pharmacy clerk staffing increased by 25.37%. Between 1989 and 1998, pharmacist staffing in U.S. medical-surgical hospitals increased at less than half the rates for registered nurses and total hospital personnel. PMID- 10832498 TI - Pharmacist job turnover, length of service, and reasons for leaving, 1983-1997. AB - Pharmacist job turnover from 1983 to 1997 was studied. Data were collected from a randomized 1997 mail survey of 1600 licensed pharmacists in four states (Ohio, Massachusetts, Oregon, and Alabama). The survey instrument included questions on pharmacist demographics, work schedules, salary, and work history. A total of 541 pharmacists responded, yielding an adjusted response rate of 34.5%. Information was provided on a total of 1697 jobs with start dates from 1931 to 1997. Pharmacist job turnover was fairly steady across the 1983-1997 period, averaging 11% annually. The average median tenure of pharmacists who left jobs was 32 months. The percentage of pharmacists leaving jobs and ranking stress as the reason for leaving increased, and the percentage of leavers ranking salary as the reason decreased. Women had a significantly higher annual turnover rate (15%) than men (9.7%), and they stayed in jobs for significantly less time (25.2 months) than men (56.5 months). There were no differences in turnover rates across practice settings. A larger percentage of pharmacists leaving jobs in large chain and institutional settings ranked stress as a reason for leaving than pharmacists leaving independent or small chain pharmacies. A larger percentage of pharmacists leaving independent or small chain pharmacies ranked salary as a reason than pharmacists in the other two settings. Pharmacist job turnover averaged 11% per year between 1983 and 1997. Pharmacists who left jobs typically stayed less than three years. The percentage citing stress as a reason for leaving increased, and the percentage citing salary decreased. PMID- 10832499 TI - Factors limiting home infusion therapy in Japan. PMID- 10832500 TI - Stability of meropenem in a portable infusion device in a cold pouch. PMID- 10832501 TI - Blurred vision from ipratropium bromide inhalation. PMID- 10832502 TI - Losartan-induced cough after lisinopril therapy. PMID- 10832503 TI - Improvements in delivering drug information to elderly patients. PMID- 10832504 TI - Perplexed about ASHP council's perspective on assisted-suicide policy. PMID- 10832505 TI - Pharmacists' perceptions of a therapeutic-interchange program. PMID- 10832506 TI - Effect of Sahaja yoga meditation on auditory evoked potentials (AEP) and visual contrast sensitivity (VCS) in epileptics. AB - The effect of Sahaja yoga meditation on 32 patients with primary idiopathic epilepsy on regular and maintained antiepileptic medication was studied. The patients were randomly divided into 3 groups: group I practiced Sahaja Yoga meditation twice daily for 6 months under proper guidance; group II practiced postural exercises mimicking the meditation for the same duration; and group III was the control group. Visual Contrast Sensitivity (VCS), Auditory Evoked Potentials (AEP), Brainstem Auditory Evoked Potentials (BAEP), and Mid Latency Responses (MLR) were recorded initially (0 month) and at 3 and 6 months for each group. There was a significant improvement in VCS following meditation practice in group I participants. Na, the first prominent negative peak of MLR and Pa, the positive peak following Na did not register changes in latency. The Na-Pa amplitude of MLR also showed a significant increase. There were no significant changes in the absolute and interpeak latencies of BAEP. The reduced level of stress following meditation practice may make patients more responsive to specific stimuli. Sahaja Yoga meditation appears to bring about changes in some of the electrophysiological responses studied in epileptic patients. PMID- 10832507 TI - Self-regulation of slow cortical potentials in children with migraine: an exploratory study. AB - Migraine patients are characterized by increased amplitudes of slow cortical potentials (SCPs), representing pronounced excitability of cortical networks. The present study investigated the efficiency of biofeedback training of SCPs in young migraineurs. Ten children suffering from migraine without aura participated in 10 feedback sessions. They were compared with 10 healthy children for regulation abilities of cortical negativity and with 10 migraineurs from the waiting list for clinical efficacy. During the first two sessions, the migraine children were characterised by lacking ability to control cortical negativity, especially during transfer trials, compared with healthy controls. However, there was no difference following 10 sessions of training. Feedback training was accompanied by significant reduction of cortical excitability. This was probably responsible for the clinical efficacy of the training; a significant reduction of days with migraine and other headache parameters was observed. It is suggested that normalization of the threshold regulation of cortical excitability during feedback training may result in clinical improvement. PMID- 10832508 TI - Reliability and stability of contingent negative variation. AB - Reliability parameters of a test indicate the stability (and quality) of the test itself. Reliability coefficients greater than 0.70 suggest an attribute as being sufficiently stable over time to be characterized as a trait. Reliability parameters of contingent negative variation (CNV) amplitudes in 27 healthy individuals were determined using a test-retest design. CNV was recorded at Cz, with an interstimulus interval of 3 s, on 2 separate occasions: initial session and 10 days later. Correlation coefficients between the 2 recording sessions were 0.675 for the total-CNV (tCNV), 0.855 for the early component (iCNV), 0.631 for the late component (lCNV), and 0.420 for the post-imperative negative variation (PINV). Statistical retest parameters for Spearman Brown were 0.806 for tCNV, 0.922 for iCNV, 0.774 for lCNV, and 0.655 for PINV. The iCNV, more than the other parameters, remained stable over the period of 10 days. It is suggested that the described standardized CNV recording procedure ensures reproducible and stable results in healthy subjects. PMID- 10832509 TI - Can taste aversion prevent bruxism? AB - The first part of this note sketches a biofeedback modality for the treatment of bruxism. A mildly aversive, safe liquid is inserted into, and sealed in, small, bilaterally sleeved, polyethylene capsules. Two capsules are attached to a simple dental appliance that comfortably and securely places them between the lower and upper back teeth. The appliance and capsules are worn at night or at other times when bruxism is suspected to occur. Whenever a sleeping or an awake patient attempts to brux, one or both capsules rupture and the liquid is released into the mouth. The liquid then draws the patient's conscious attention to, and forestalls, any attempt of teeth clenching or grinding. Variations of the method and device can be used to diagnose bruxism. The second part of this note describes the long-term application of this taste-based approach to one chronic bruxer. PMID- 10832510 TI - Use of microneurography to evaluate sympathetic activity in hypertension: a brief review. AB - Muscle sympathetic nerve activity (MSNA) is an important variable in the study of autonomic activity in both normotensive and hypertensive subjects. It is measured directly from the peroneal nerve using microneurography. The technique is complex and difficult to learn, but yields accurate and direct information about sympathetic nerve impulses. MSNA provides not only greater reproducibility than other measures of sympathetic activity, but also a clearer and more consistent reflection of changes in sympathetic activity caused by changes in the subject's status or disease. This technique has been used primarily in basic research settings studying stress and hypertension. It has much potential to enhance our understanding of sympathetic nervous system activity and its role in applied psychophysiology and biofeedback. PMID- 10832511 TI - Socio-economic differences in foundation-level literacy. AB - The foundation literacy skills of children from differing socio-economic backgrounds were investigated in a cross-sectional study. The children were aged between 4 and 8 years and attended Nursery or Primary 1, 2 or 3 classes. Low socioeconomic status (SES) was associated with impairments for chronological age in letter knowledge as well as in both logographic and alphabetic foundation components. There was also an effect on metaphonological++ skill. However, once the SES groups were equated for reading age, high and low SES performance was indistinguishable. The results suggest that delayed acquisition of foundation literacy skills is traceable to a delay in acquiring letter-sound knowledge. Implications for intervention are discussed in the context of the foundation literacy framework. PMID- 10832512 TI - Real age of acquisition effects in word naming and lexical decision. AB - Age of acquisition (AoA) has been reported to be a predictor of the speed of reading words aloud (word naming) and lexical decision, with early-acquired words being responded to faster than later-acquired words in both tasks. All previous studies of AoA effects have, however, relied upon adult estimates of word learning age the validity of which it is easy to cast doubt upon. Using objective age of acquisition norms derived from children's naming data, this study shows that AoA effects do not depend upon the use of adult ratings. In addition to effects of real AoA, influences of word frequency and orthographic neighbourhood size were obtained in both word naming and lexical decision. Imageability affected lexical decision but not word naming, while the characteristics of the word's initial phoneme affected word naming but not lexical decision. PMID- 10832513 TI - Working memory deficits in current and previous users of MDMA ('ecstasy'). AB - Current and previous users of the drug MDMA ('ecstasy') were tested on measures of central executive functioning, information processing speed, and on self report measures of arousal and anxiety. The results were compared with those for a control group who did not use MDMA. Relative to the control group, both user groups were found to be impaired in some aspects of central executive functioning. Also, there were significant group differences on the measures of anxiety (users were more anxious) and on arousal (previous users scoring higher on the arousal measure relative to current users). Users processed information as quickly as non-users but less accurately. Some possible mediators of the above group differences are discussed. PMID- 10832514 TI - Memory conformity: exploring misinformation effects when presented by another person. AB - Two experiments demonstrate that post-event information, when delivered by another person, can affect people's memory reports. In the first experiment participants were shown several cars, and later, in pairs, given an 'old'/'new' recognition test on these cars plus several lures. There was a small but reliable effect of memory conformity. When the person was given misinformation this lowered accuracy, while presenting accurate information increased accuracy. In the second experiment participants, in pairs, viewed an identical crime except that half saw an accomplice with the thief and half did not. Initial memories were very accurate, but after discussing the crime with the other person in the pair (who saw a slightly different sequence), most pairs conformed. Confidence ratings strongly predicted which person in the pair persuaded the other. Parallels with eyewitness testimony in the Oklahoma bombing case and implications for police interviewing more generally are discussed. PMID- 10832515 TI - A conversation-based process tracing method for use with naturalistic decisions: an evaluation study. AB - This study is an evaluation of a process tracing method developed for naturalistic decisions, in this case a consumer choice task. The method is based on Huber et al.'s (1997) Active Information Search (AIS) technique, but develops it by providing spoken rather than written answers to respondents' questions, and by including think aloud instructions. The technique is used within a conversation-based situation, rather than the respondent thinking aloud 'into an empty space', as is conventionally the case in think aloud techniques. The method results in a concurrent verbal protocol as respondents make their decisions, and a retrospective report in the form of a post-decision summary. The method was found to be virtually non-reactive in relation to think aloud, although the variable of Preliminary Attribute Elicitation showed some evidence of reactivity. This was a methodological evaluation, and as such the data reported are essentially descriptive. Nevertheless, the data obtained indicate that the method is capable of producing information about decision processes which could have theoretical importance in terms of evaluating models of decision-making. PMID- 10832516 TI - The structure of the Eysenck Personality Profiler. AB - The dominant issue in personality research over the last decade has been concerned with the fundamental structure of personality and the best measures of that structure. Exploratory factor analysis was used to investigate possible three- and five-factor solutions to the Eysenck Personality Profiler (EPP; Eysenck, Barrett, Wilson, & Jackson, 1992) which consists of 21 primary scales categorized under three super-factors. Little evidence was found to support Costa and McCrae's (1995) unequivocal comment that a five-factor solution fitted the data well. Confirmatory factor analysis was also used, by means of structural equation modelling, to estimate the goodness of fit of three- and five-factor models and little evidence was found to favour one solution over the other. A shorter version of the EPP, which consists of just nine scales, seemed to favour a three-factor solution. Various criticisms of the EPP are also made: some scales have relatively low alpha, there seem to be too many neuroticism scales and the three category response scales seem less than ideal. PMID- 10832517 TI - The role of left-right symmetry in the encodement of spatial orientations. AB - Three experiments investigated a hypothesis, suggested by studies of the difficulties of discriminating between shapes forming symmetrical pairs, that spatial orientations of thin flat plates (lamellae) may be encoded in a plane, the encodement consisting of two enantiomorphs. The results indicated that participants encoded the spatial orientation of lamellar stimuli in terms of the difference in cogency between their two enantiomorphic elements (Expt 1). The difference in the cogency of the two enantiomorphs is related to the orientation of the plane containing the lamellar stimulus with respect to the participant's fronto-parallel plane (Expt 2). The two possible orientations of a lamella which yield the same difference of cogency, but which differ in spatial orientation (e.g. lamella 'b' set at 30 degrees or set at 150 degrees) are distinguished by the manner in which the two enantiomorphic elements are arranged with respect to their axis of symmetry (Expt 3). The results suggest that the orientation of a lamella may be encoded as a two-dimensional representation and hence that three dimensions may be encoded by two by means of enantiomorphs. Implications of this finding for the encodements of three-dimensional solids, wherein pronounced contours may fulfil the same role as do the edges of lamella, are discussed briefly. PMID- 10832518 TI - When will the ball rebound? Evidence for the usefulness of mental analogues in appraising the duration of motions. AB - This study explores the way people appraise the duration of motions when these are only seen momentarily. Three possible theoretical accounts of this skill were developed. The first and second held an analogy between the duration of motions and their visuospatial representations. The third account was based on the existence of a pure chronometrical representation. Three copies of a tennis ball trajectory were created to reproduce the characteristics of the trajectory in terms of each account. The participants first memorized the copy and then used it to appraise the duration of the actual ball trajectory seen only momentarily at the beginning of its flight. The performance of a group given the visuospatial copy paralleled those of a referent group who were allowed to see the actual referent trajectory to memorize its duration. Results showed the importance of visuospatial analogous representations. However, a spatial representation seems to allow better memorization of motion duration. PMID- 10832519 TI - Jean Theophile Desaguliers (1683-1744) and eighteenth century vision research. AB - The emergence of psychology as an empirical discipline was influenced to a great extent by experimental investigations of visual phenomena, particularly in the nineteenth century. Less attention has been paid to experimental enquiries conducted in the eighteenth century, especially those of Jean Theophile Desagliers (1683-1744). He was an ardent advocate of Newtonian optics, on which he lectured and gave demonstrations. His research on colour and binocularity is outlined, together with those of other students of vision in that century. Experiments on visual vertigo conducted at the end of the century are also described. In 1716 Desaguliers reported a method of binocular combination that became widely employed in other studies of binocular vision, namely, placing an aperture in such a position that two more distant, adjacent objects were in the optical axes of each eye. Under these circumstances red and green patches of silk did not mix after the manner of combining prismatic lights, but engaged in rivalry. Desaguliers also investigated size perception and showed that apparent size was determined by apparent distance rather than physical distance. Moreover, he did not base his conclusions on his own observation but on those of 'any unprejudic'd Person'. Thus, both stimulus control and the use of the unbiased observer were employed in eighteenth century experimental studies of vision. PMID- 10832520 TI - Dalzell's theorem and the analysis of proportions: a methodological note. AB - The golden section is a well-known proportion that occurs when something (e.g. a line) is divided into two unequal parts such that the smaller (m) is to the larger (M) as the larger is to the sum of the two (i.e. m/M = M/(M + m) = .618). Dalzell's theorem holds that the absolute value of the difference between M/(M + m) and .618 will tend to be smaller than the corresponding difference between m/M and .618. This means that the use of M/(M + m) ratios leads to results that are more supportive of the golden section hypothesis than does the use of m/M ratios. Notice that M/(M + m) corresponds to the proportion of Ms that will occur; while m/M corresponds to the odds that m will occur. While these are mathematically equivalent, in practice they may lead to different interpretations of the same data. Although originally envisaged as applying to the golden section, Dalzell's theorem may have implications for any study that uses either a proportion or the odds as a dependent measure. The use of proportions may produce results that are closer to a predicted value than will the use of the odds as a dependent measure. PMID- 10832521 TI - [At the beginning of the third millennium: what kind of surgery, what kind of surgeon?]. AB - Technological progress and the evolution of biological knowledge lead us to expect major developments in the fields of organ replacement, minimally invasive surgery and tumour therapy in the coming decades. It is widely believed that xenotransplantation may offer a possible solution to the problem of the current chronic shortage of donors, though their use at present encounters formidable immunological obstacles, particularly as regards discordant transplants. Genetic engineering and the realisation of transgenic donors as well as new strategies aimed at modifying the immune status of the donor may well be the key to new developments in this sector. Selective culturing of tissues and the production of totally implantable artificial organs are additional lines of research which may provide useful means of meeting the increasing demand for organs. The enormous advances made in the field of high-tech solutions applied to the biomedical disciplines are destined to open up new frontiers in diagnostics, operating technique and the permanent training of surgeons. In particular, we are about to witness an extension of the use of computer technology and robotics, embracing the simulation and reproduction of virtual reality, telemedicine (teleconsulting and telesurgery) and the creation of experimental models. Minimally invasive surgery will unquestionably benefit from this progress with the possibility of extending its range of indications to procedures in which human limitations are more evident owing to difficulties of access and the size of the target, at the same time guaranteeing high-precision manoeuvres. In the oncological field, high expectations accompany the advances in our knowledge of molecular biology with the genetic mapping of tumours which may pave the way for screening programs and gene therapy. Alongside the scientific and technological development of the surgery of the future, we can expect to see the increasing emergence of: a) economic problems related to the progressive increase in healthcare costs, amongst other things as a result of the powerful impact of high-tech solutions; b) social problems in terms of the inevitable conflict between cost-containment policies and the right to healthcare; c) moral and bioethical problems relating to the possible establishment of genetic registries and to the different patient care provider relationship which is likely to come about as the result of the interference of machines or robots. The surgeon's persona may undergo profound changes, not merely from the technical and professional training standpoint, but also with regard to his or her effective role, which may no longer be that of a central, dominant figure, but may be scaled down in a context of multidisciplinary co-operation. PMID- 10832522 TI - Topographic diagnosis and surgical treatment of insulinoma. AB - AIM: Controversy continues to reign with regard to the need for preoperative localization of insulinomas and to which are the most sensitive and accurate diagnostic imaging modalities. Our aim was to determine the role of diagnostic procedures and suggest which of them are really useful. METHODS: Over a 12-year period 34 patients underwent several preoperative diagnostic procedures to localize the insulinoma: ultrasonography (US) in 20 cases, computed tomography (CT) in 26, magnetic resonance imaging (MRI) in 28, selective angiography in 8, arterial stimulation venous sampling (ASVS) in 23 and Octreoscan in 26. All patients underwent surgical palpation and in 32 cases intraoperative ultrasonography (IOUS) was performed. Twenty-six cases underwent enucleation, six had distal pancreatic resections and two patients had only exploratory laparotomy with liver biopsies. We compared the findings of the diagnostic procedures and analyzed the surgical treatment chosen according to the pancreatic site of the tumor. RESULTS: In 32 (94.1%) of the 34 patients with clinically suspected pancreatic insulinoma the tumor was found at surgery. Preoperative US achieved 5.2% sensitivity, CT 29.1%, selective angiography 28.5% and MRI 76.9%. ASVS achieved 91.3% sensitivity and diagnostic accuracy whereas Octreoscan achieved only 65.3% diagnostic accuracy. Surgical palpation performed before IOUS identified the tumors in 30/34 patients: in the other four cases, one was a false positive result (a cyst in the pancreatic head), two were true negatives and one was a false negative. Surgical palpation therefore yielded 88.2% diagnostic accuracy. IOUS was performed in 32 cases and localized the tumors in 29/32 cases (sensitivity: 96.6%) with one false-negative result (diagnostic accuracy: 96.8%). The operative mortality was 2.9% and the morbidity 24.6% (30.7% in patients treated by tumor enucleation). CONCLUSIONS: No single diagnostic imaging modality is reliable for localizing insulinoma. We therefore suggest combined MRI, ASVS and IOUS. ASVS provides particularly useful information for planning manual palpation and intraoperative ultrasonography. PMID- 10832523 TI - [Laparoscopic versus conventional surgery in the treatment of colorectal diseases]. AB - The aim of the study was to compare the results obtained with laparoscopic (LPS) and laparotomic (LPT) colorectal resection after our initial experience with the laparoscopic technique. Fifty-six patients were submitted to colorectal resection, 26 with the LPS and 30 with the LPT technique. Eighteen patients out of 26 in the LPS group and 22/30 in the LPT group had malignancies. All resections were performed with a curative intent. The mean operating time was 220 min in the LPS group and 208 min in the LPT group. Mean blood loss was 287 ml and 312 ml, respectively (blood transfusions were needed in 1/26 and in 7/30 patients). The rates of major complications were 9.5% and 5.7%, respectively. There was no mortality. The conversion rate for the LPS group was 19.2%. In the cancer patients, no significant difference was observed between the two groups as regards postoperative staging. The mean length of specimens and the mean distance of the tumours from the resection margins were adequate. The mean number of lymph nodes harvested was 11.8 in the LPS group as against 18.5 in the LPT group. No early recurrences were observed. Resumption of gastrointestinal function was faster in the LPS patients who underwent the surgical procedure under general anaesthesia associated with epidural anaesthesia/postoperative analgesia. In conclusion, these preliminary results indicate that laparoscopic colorectal surgery is feasible and that the resections in cancer patients appear to be oncologically adequate. Long-term follow-up is needed for reliable assessment of oncological outcomes. PMID- 10832524 TI - Intraoperative and percutaneous radiofrequency thermal ablation in the treatment of hepatocellular carcinoma. AB - AIMS AND BACKGROUND: The aim of the study was to evaluate feasibility, survival rate, complications and length of hospital stay in 47 patients with hepatocellular carcinoma (HCC) treated by radiofrequency thermal ablation (RFTA). Though the treatment of choice for HCC is surgical resection, the strong association of this disease with cirrhosis often rules out this procedure. Many investigations have been conducted in order to identify alternative therapies. Preliminary studies of radiofrequency thermal ablation have shown that the technique is effective and safe, achieving a predictable area of tumor tissue coagulative necrosis and sparing the surrounding cirrhotic parenchyma, without any significant side effects. In addition, this technique, which can be performed percutaneously, allows very short hospital stays. PATIENTS AND METHODS: We report the results of a series of 47 cirrhotic patients with 52 HCC nodules (mean diameter 2.9 cm, range 1-6 cm) treated in our Institute between May 1997 and June 1999 by RFTA using an expandable needle with four hooks at its tip. All patients had hepatic cirrhosis (32 Child A, 13 Child B and two Child C). We treated patients with both unifocal (35 patients) and multifocal HCC (12 patients); 33 patients underwent percutaneous RFTA (54 passes), while in 14 cases RFTA was performed during laparotomy (22 passes). RESULTS: The mean number of passes to achieve complete necrosis was 1.43 in 28 patients with unifocal HCC treated by percutaneous RFTA, 1.7 in 7 patients with unifocal HCC treated by intraoperative RFTA, 2.8 in 5 patients with multifocal HCC treated by percutaneous RFTA and 1.43 in 7 patients with multifocal HCC treated by intraoperative RFTA. No deaths related to the procedure or major complications occurred. Post-treatment dynamic CT was performed in all patients. All patients but one were followed-up for a mean period of 11.8 months (1-25 months). Six patients died during the follow-up (three Child A, two Child B and one Child C. The actuarial survival, computed by the Kaplan-Meier method, was 83% at 24 months. The mean hospital stay was 3.4 days in patients treated by percutaneous RFTA and 11.2 days in those treated by intraoperative RFTA. CONCLUSIONS: In our opinion RFTA is an effective, safe technique capable of achieving good results in the conservative therapy of small HCC. We believe that curative ablation is possible for HCC nodules measuring up to 3 cm in diameter. Further studies of longer duration are necessary. PMID- 10832525 TI - [Benign thyroid disease: 20-year experience in surgical therapy]. AB - This retrospective study compares recurrence and postoperative complication rates after isthmo-lobectomy and subtotal thyroidectomy (group I) vs near-total and total thyroidectomy (group II) for benign thyroid disease. Seven hundred and forty-three patients were operated on for thyroid diseases over the period from 1977 to 1998. We considered 202 patients operated on for benign thyroid disease from 1988 to 1998. The follow-up ranged from 1 to 10 years (mean: 3.4 yrs). One hundred and thirty-two patients (65.3%) were operated on for bilateral nodular goitre, 35 (17.3%) for unilateral nodular goitre, 14 (6.9%) for toxic goitre and 21 (10.4%) for thyroiditis. Over the period 1988-1992, 19 patients underwent isthmo-lobectomy and 71 subtotal thyroidectomy (group I). From 1993 to 1998, 39 patients underwent near-total thyroidectomy and 61 total thyroidectomy (group II). The relapse rate was 14.4% in group I, while there were no recurrences in group II (p = 0.000064). Temporary hypocalcaemia was significantly higher (p = 0.000001) in group II (29%) than in group I (2.2%). Within group II, the rate was significantly higher (p = 0.0013) after total thyroidectomy (37.7%) than after near-total thyroidectomy (15.4%). In our experience, near-total and total thyroidectomy are an appropriate approach for preventing recurrence in patients with benign thyroid disease despite the fact that the risk of temporary hypocalcaemia is higher than after less radical surgery. Near-total thyroidectomy and the exercise of all due care in the surgical technique may help to reduce its incidence. PMID- 10832526 TI - [Surgical treatment of differentiated carcinoma of the thyroid. Follow-up of 99 cases]. AB - The surgical treatment of differentiated thyroid carcinoma continues to be a matter of considerable debate in terms of defining the appropriate extent of thyroid or lymph node resection to ensure optimal patient survival. Whereas, at organ level, the majority of surgeons are in favor of total thyroidectomy, both the extent and timing of lymphadenectomy remain controversial issues. In the light of this, the Authors have conducted a retrospective study in 99 consecutive patients with differentiated thyroid carcinoma. As regards the distribution of the cancers in terms of TNM staging, 60 were stage I, 27 stage II, 11 stage III and 1 stage 4. Almost all the patients underwent total thyroidectomy. Lymphadenectomy was performed at the same time as thyroidectomy in papillary cancers when the nodes were clinically palpable and at a later date in those cases where the nodes subsequently became palpable at follow-up. The in-hospital mortality was 3% and was unrelated to the operation. The median follow-up was 95.8 months (7.98 years). Sixty-nine patients are still alive (71.8%), 66 of them disease-free (68.7%) and 3 with lymph node metastases (3.1%). Our results and those of other investigators suggest that total thyroidectomy should be the first therapeutic choice in differentiated cancers of the thyroid, but allow us to draw no firm conclusions regarding the controversial issue as to which type of lymphadenectomy can best ensure patient survival. To solve this problem we believe that multicenter randomized trials will be necessary. However, progress in molecular biology and tumor genetics is likely to enable us to identify new prognostic factors which may prove useful when deciding on the most appropriate therapeutic option. PMID- 10832527 TI - [Original reconstruction technique after duodenopancreatectomy]. AB - The authors present an original reconstruction technique after pancreaticoduodenectomy, with anastomosis between the pancreatic stump and the posterior wall of the stomach, using two Roux-en-Y loops to separate the hepaticojejunostomy from the pancreaticogastrostomy and gastrojejunostomy in order to reduce postoperative complications and mortality. Eighteen consecutive patients underwent the procedure. There was no mortality and no pancreaticogastrostomy leaks occurred. Two (11.1%) gastric bleeds occurred in the first two cases. Twelve cases (66.6%) presented alimentary emesis on postoperative day 5 or 6 after food intake. Three patients (16.6%) had postoperative diarrhea. There were no complications calling for reoperation. The mean hospital stay was 14.4 days. No significant late complications were observed. The procedure is easy and safe with no mortality and with one of the lowest complication rates in the literature. PMID- 10832528 TI - [Sensitivity and specificity of transrectal ultrasonography in the preoperative staging and postoperative follow-up in rectal neoplasms. Experience with 100 clinical cases]. AB - Preoperative staging plays an important role in the surgical treatment of rectal cancer. The most sensitive imaging techniques currently available are CT, MRI and transanal ultrasound (TAUS). The aim of the study was to evaluate the sensitivity and specificity of TAUS in the preoperative staging and postoperative follow-up of rectal cancer. METHODS: From January 1992 to May 1999, TAUS was used to study 100 patients with rectal cancer. RESULTS: Patients were staged according to the Astler-Coller classification: 1) A: 8 patients; 2) B1: 16 patients; 3) B2: 22 patients; C1: 30 patients; C2: 24 patients. The sensitivity and specificity of TAUS in the preoperative staging of these tumors were 96% for the T parameter, and 100% for the N parameter. The N but not the T parameter results are in line with the values reported in the international literature. CONCLUSION: Transanal ultrasound, in our personal experience, has proved to be a very accurate imaging technique in the preoperative staging and postoperative follow-up of rectal cancer. PMID- 10832529 TI - [Development of anesthesiological techniques in the surgical treatment of inguinal hernia]. AB - The Authors describe the evolution of their anesthesiological techniques in the surgical repair of inguinal hernia. In this study they compare indications, complications, costs, hospital stay, length of surgery, postoperative pain and return to work after operations performed under local, spinal and general anesthesia. In their experience inguinal hernia treatment with local anesthesia and a tension-free technique is the preferred method of surgical repair. PMID- 10832530 TI - Gastric neurofibroma in a patient with von Reklinghausen's disease: a cause of upper gastrointestinal hemorrhage. AB - Gastric neurofibroma associated with von Reklinghausen's disease is a rare clinical entity. We report a case of gastric neurofibroma with gastrointestinal bleeding and severe anaemia. Surgical resection is the treatment of choice. The recommended follow-up is an annual complete cell blood count and stool testing for occult blood. PMID- 10832531 TI - [Synchronous neoplasms of the colon and kidney: analysis of 2 case reports]. AB - Up until only a few decades ago synchronous renal cell carcinoma associated with primary tumors of other organs was diagnosed almost exclusively in necropsy series. Recently the widespread use of ultrasonography and CT has permitted diagnosis of clinically silent renal cell carcinoma in patients undergoing a work up for other primary tumors. We report two cases of synchronous colon and renal cancer: 1) a 75 year old woman presented bilateral pulmonary nodules at chest X Ray and stratigraphy. Total-body CT scan showed multiple, apparently metastatic, bilateral pulmonary lesions and two tumors in the right kidney and ascending colon. A right nephrectomy and right hemicolectomy were performed. She succumbed after 18 months as a result of metastatic spread to the liver, lungs and bone; 2) a 64 year old man was discovered to have a colon cancer during a barium enema examination and endoscopy. CT scan of the abdomen yielded an incidental diagnosis of a tumor in the left kidney. A left nephrectomy and left hemicolectomy were performed. After 6 months CT revealed no evidence of recurrence or metastases. Histology findings revealed two primary malignancies in both cases: a clear cell renal carcinoma and an adenocarcinoma of the colon. PMID- 10832532 TI - [Appendiceal oxyuriasis: a case report and review of the literature]. AB - The authors describe a case of appendicular oxyuriasis and then go on to review the international literature on this condition. Nothing in the case report provides the surgeon with useful pointers which may help him distinguish between appendicular oxyuriasis and common appendicitis. This is in line with all the existing literature which yields no elements capable of suggesting an accurate preoperative diagnosis. Only anamnestic evidence of previous oxyuriasis can help the clinician in this direction. It should be noted, however, that there are no significant differences in postoperative morbidity between "common" acute appendicitis and appendicitis due to oxyuriasis, when the parasitosis is adequately treated. PMID- 10832533 TI - [Transverse minilaparotomy as an access route in right colon disease: a valid alternative to midline laparotomy]. AB - AIMS OF THE STUDY: 1. To evaluate the feasibility and appropriateness of transverse right minilaparotomy for right colectomy and lymphadenectomy. 2. To identify the clinical advantages, if any, of this access route compared to the more traditional midline incision. PATIENTS: 44 patients with right colon adenocarcinoma undergoing colonic resection via a transverse or midline laparotomy. METHODS: The transverse incision starts 1 cm above the umbilicus, running from the midline to the anterior axillary line. RESULTS: 17 (39%) transverse minilaparotomies and 27 (51%) midline laparotomies were performed, respectively. The mean operative time was 107 min for patients with the transverse incision and 157 min. for those with midline laparotomy. Twelve out of 17 patients (70%) with the transverse incision experienced no or only mild pain as against 11 out of 27 patients (41%) with midline laparotomy. Bowel function was slightly faster in the first group of patients, though the difference was not statistically significant. Forty-seven percent of patients with transverse minilaparotomy were discharged within 8 days postoperatively as against 4% with midline laparotomy (P = 0.001). No major complications occurred in the first group, while 2 patients (7%) operated on via the midline laparotomy experienced complications (haemorrhage, deep venous thrombosis). CONCLUSION: Although this is a preliminary study in only 44 patients, our results support the idea that transverse minilaparotomy is a less traumatic access route to the right colon, in addition to offering an adequate exposure of the operative field, faster postoperative recovery and a shorter hospital stay. We believe it to be a good alternative to midline laparotomy for right colon surgery. PMID- 10832534 TI - [Historical evolution of hepatobiliopancreatic surgery]. PMID- 10832535 TI - The impact of an incomplete preoperative colonoscopy in patients with colorectal cancer. AB - Total large bowel evaluation remains an essential step in the treatment of patients with colorectal cancer (CRC). Colonoscopy is the gold standard in the evaluation of the colon for colorectal tumors, but may be incomplete due to tumor obstruction, which is a frequent event in distal cancers. Double-contrast barium enema has a lower accuracy and is not ideal in the presence of signs of obstruction. In theory, intraoperative colonoscopy is a valid alternative, but its routine use is impeded by various practical limitations. Preoperative survey of the colon in 521 consecutive patients treated for CRC in our department was based on colonoscopy (92.5%). Our series was characterised by a high percentage of distal lesions (76.4%) and therefore by a high percentage of incomplete preoperative colonoscopies (50.4%) due to tumor obstruction. In the presence of an incomplete preoperative colonoscopy, we evaluated the entire colon with a double-contrast barium enema in selected cases (36.7%) and with a postoperative colonoscopy within 3 months of surgery in almost all patients (93.4%). The overall rate of complete endoscopic evaluation, either pre- or postoperatively, was 96.7%. The incidence of adenomas was significantly higher in the preoperative examinations as compared to early postoperative colonoscopy. This means that in distal cancer the vast majority of polyps will be located in the distal colon and therefore included in a standard resection. Four patients (0.8%), required a second operation for treatment of a missed lesion (2 benign and 2 malignant). The need for a repeat surgery apparently did not affect the therapeutic results in these four patients. On the basis of our experience, intraoperative colonoscopy would not appear to be a mandatory procedure in all cases of incomplete preoperative evaluation of the colon. However, in the absence of prospective, randomised trials comparing intraoperative vs early postoperative colonoscopy, the dilemma as to the strategy of choice remains. PMID- 10832536 TI - [New developments in Crohn's disease: solution of doctrinal mysteries and reinstatement as a surgically treatable disease. 1. The process is not a form of enteritis but lymphedema contaminated by intestinal contents]. AB - INTRODUCTION: It is unanimously acknowledged that the aetiopathogenesis of Crohn's disease (CD) is still a mystery despite the fact that almost 70 years have elapsed since it nosological identification. Another mystery and major problem of CD of the bowel is its histopathology, which has yet to be clarified; consequently, we do not know what kind of disease it is, that is to say, which process, among the ones known to us, it actually corresponds to. MATERIAL AND METHODS: The present paper presents the results of a new histopathological interpretation of the disease on the basis of 100 surgical cases of Crohn's ileitis and ileocolitis. RESULTS AND CONCLUSIONS: The disease is not a form of enteritis, but a lymphoedema of the intestinal wall, caused by a congenital lack of patency of the mesenteric lymphatic collectors, giving rise to stasis of lymph in the lymphatic network of the corresponding bowel segment and consequently to hypertension which dilates the lymph vessels in the intestinal wall. The lymph, which cannot be reabsorbed by the veins, because the venous wall denies entry to protein macromolecules, accumulates in the interstitial space with the result that the intestinal wall thickens considerably. Since the process takes place in the intestine perfused by a continuous flow of septic matter, the process is complicated by the development of lymphocytic lymphangitis, which occurs throughout almost the entire lymphatic network, obstructing the dilated lumen, limiting the dispersion of lymph and preventing gross spread of the intestinal contents into the dilated lymphatic vessels. The lymphocytic lymphangitis causes "transparietal phlogosis", which is rightly regarded as a characteristic of the disease, but is wrongly interpreted by those who have studied it. This transparietal phlogosis is neither excessive nor disproportionate to its task of obstructing the intestinal contents; it is transparietal because the lymphatic network where it takes places is itself transparietal. The reactive lymphocytic lymphangitis prevents the gross spread of the intestinal contents into the dilated lymphatic network, as stated above, but cannot prevent the limited uptake of dissolvable toxic bacterial substances (FMLP, PG-PS, LPS) conjugated in immuno complexes. The immuno-complexes developing in the lymphatic network undergo lymph and blood-borne spread, causing; obstructive lymphangitis in branches of the mesenteric lymphatic collectors which are still patent, increasing the degree of lymphatic obstruction and thus the extent of the intestinal lesions; general complications of so-called "metastatic Crohn's disease", which were once interpreted as autoimmune processes, but actually correspond to forms of vasculitis which may be localised in any of the organs or apparatuses of the body, sometimes proving extremely serious. Lymph dispersion in the intestinal lumen, causes not only diarrhoea, but often also lymphocytopenia and hypoproteinaemia, and therefore an immunodepression syndrome, which in the long term may give rise to intestinal and extraintestinal cancer. PMID- 10832537 TI - [Evaluation of risk factors in the development of postoperative complications in patients undergoing liver resection for cancer]. AB - A consecutive series of 134 hepatic resections for primary and metastatic cancer were analyzed to identify the risk factors for post-operative complications in patients with and without impaired liver reserve. Between January 1992 and January 2000 were performed 55 hepatectomies (41%--group 1) in 54 cirrhotic patients for hepatocarcinoma and 79 hepatic resections (59%--group 2) in 66 patients for primary hepatic malignancies or metastatic liver tumours in non cirrhotic liver. Among major postoperative complications bile leakage was recorded in 8 patients (6%) (6% with impaired liver reserve and 6% with normal reserve), hepatic failure in 8 patients (6%) (9% vs 4%; P = NS), ascites in 7 patients (5%) (11% vs 1%; P = 0.01), pneumonia in 4 patients (3%) (5% vs 1%; P = NS), intra-abdominal abscess in 2 patients (1%) (2% vs 1%; P = NS), postoperative haemorrhage in 2 patients (1%) (4% vs 0; P = NS), and gastrointestinal bleeding in 2 patients (1%) (4% vs 0; P = NS). There were 6 perioperative deaths (4%) (7% vs 2%; P = NS). The mean hospital stay was 21 +/- 10 days (range: 5-57) (24 +/- 10 vs 20 +/- 10; P = 0.02). Liver resection is a safe procedure even in cirrhotic patients providing they are well selected and there is minimal intraoperative blood loss. PMID- 10832539 TI - [Intrathoracic goiter: experience with 61 surgically treated cases]. AB - Sixty-one patients with substernal goiter--42 women and 9 men (female:male ratio 2.2:1), mean age 57.5 years--underwent surgical resection in the General and Thoracic Surgery Division of the Surgery Department of the University of Catania from January 1980 to March 1999. Six patients (9.8%) had previously undergone cervicotomy. Forty-nine patients (80.3%) had symptoms (2 with acute respiratory failure); 22.9% had metabolic symptoms. Fifty-one patients (83.6%) had cervicomediastinal goiter, 8 (13.1%) migrated mediastinal goiter and 2 (3.3%) ectopic goiter. Forty-one cases (69.5%) were prevascular and 18 (30.5%) retrovascular; of the latter 5 were pre-visceral and 13 retrovisceral. Fifty-one thyroidectomies (6 subtotal) were performed, as well as 5 mediastinal excisions to complete removal and 4 resections confined to the mediastinal component. The 51 patients with cervicomediastinal goiter underwent cervicotomy in 42 cases (82.4%), cervicosternotomy in 7 cases (13.7%) and cervicosternothoracotomy in 2 cases (3.9%). The 8 migrated mediastinal goiters were removed via sternotomy in 6 cases, cervico thoracotomy in 1 and thoracotomy in 1. This latter approach was also used for the 2 ectopic goiters. The goiter presented signs of neoplastic degeneration in 2 cases (3.2%). Postoperative complications were: recurrent nerve palsy in 5 cases (1 bilateral definitive, 1 monolateral definitive, 3 monolateral transient), respiratory failure in 2, mediastinal hematoma in 1 and hypocalcemia in 8 (transient in 6). There was one postoperative death due to cardiorespiratory failure after cervicosternotomy. Surgical excision of substernal goiters is the best choice of treatment in view of the very low postoperative mortality and morbidity. PMID- 10832538 TI - [Liver metastasis of breast carcinoma. Results of surgical resection. Analysis of 15 operated cases]. AB - AIM OF THE STUDY: The aim of the study was to define the efficacy of liver resection for metastases from operated breast cancer by means of analysis of the results of a single institution compared to the literature data. MATERIALS AND METHODS: Over the period from 1990 to 1998, 15 patients with liver metastases from operated breast cancer were submitted to surgical resection. Thirteen patients were operated on for single metastases and two for multiple liver metastases. In the first group, a simple metastasectomy or wedge resection was sufficient, while in the second a bisegmentectomy and a right hepatectomy, respectively, were performed. Nine patients were subsequently submitted to adjuvant chemotherapy, two to chemo- and hormonotherapy, and two to hormonotherapy alone, while two other patients received no systemic treatment. RESULTS: No major complications or postoperative mortality were observed. Median survival was 44 months. The actuarial 5-year survival rate was 38.3%. Eight patients are still alive, and 7 patients have died as a result of their liver and systemic metastases. CONCLUSIONS: Hepatic metastasis from breast cancer is a good indication for surgical resection. Resection is capable of providing curative treatment and better results in terms of quality of life and survival rates, as compared to chemotherapy and hormonotherapy, especially in patients operated on for primary tumours more than three years earlier. PMID- 10832540 TI - [Thyroid nodules: comparison of preoperative and intraoperative needle aspirate and definite histological study]. AB - The need to discover malignancy is the most challenging dilemma in the management of thyroid nodules, the most common endocrine disorders, affecting 4-5% of the general population. Malignancies account for only 2-3% of cases. The aim of our study was to evaluate the predictive value of preoperative fine-needle aspiration cytology (FNAC) in surgical decision making by evaluating the final pathologic diagnosis and comparing it to the preoperative and intraoperative diagnoses. We conducted a prospective study of 30 thyroid resections. The mean age was 49 years (range: 27 to 68 years). Preoperative physical and laboratory examinations, presenting symptoms, imaging studies and predictive values of preoperative and intraoperative FNAC were analyzed. The consistency of the lesion was strongly predictive of malignancy, when "hard". Single lesions were also predictive of malignancy. The diagnostic accuracy of preoperative FNAC vs intraoperative FNAC vs frozen section histopathology was 90% vs 100% vs 96.7%; sensitivity: 91.6% vs 100% vs 100%; specificity: 90.5% vs 100% vs 94.7%, while the positive predictive value was 84.6% vs 100% vs 91.7%, and the negative predictive value 95% vs 100% vs 100%. Ultrasound-guided preoperative FNAC showed high specificity, sensitivity and accuracy in diagnosing malignancy in thyroid nodules. Intraoperative FNAC was more accurate than intraoperative frozen sections in diagnosing malignancy in thyroid nodules. PMID- 10832541 TI - [Treatment of perianal Crohn's disease]. AB - Perianal Crohn's disease (PACD) is defined as the presence of persistent lesions in the anal canal and perianal region in patients with Crohn's disease. The relative incidence of PACD in Crohn's disease patients ranges from 15 to 80% in the literature, depending on the accuracy of the clinical investigations and the clinical importance attributed to the lesions in the various study populations. The incidence is significantly higher if the intestinal disease is located in the colon-rectum rather than in the small bowel. We reviewed our experience in 105 patients with PACD, 32 of whom presenting rectal localisation of the primary disease. We observed 2 stenoses, 3 perirectal abscesses, 3 rectal ulcerations, 5 skin tags, 10 fissures and 77 fistulas. Two dilatations under narcosis, 2 intrarectal drainages of abscesses, 19 fistulotomies, 7 partial fistulotomies and insertion of loose setons, 47 loose setons and 4 anoperineal diversions were performed. The remaining patients received medical and topical treatments. After a median follow-up of 30 months, 90 patients (86%) showed a good response with improvement in functional scores, while 15 (14%) showed no improvement or a worsening requiring proctectomy in 13 cases. All patients submitted to proctectomy had rectal localization of the disease. PMID- 10832542 TI - [Pulmonary metastasis of basal cell carcinoma of the skin. 3 case reports]. AB - The Authors describe three cases of lung metastasis from a basal cell carcinoma of the skin observed in the Department of Thoracic Surgery of the National Cancer Institute in Milan. The incidence of this rare occurrence reported in the literature is approximately 0.1%, The aim of the study was to evaluate the therapeutic treatment adopted in these unusual situations. In all cases the primary basal cell carcinoma of the skin was located in the head. All three patients had several local and regional recurrences and underwent multiple surgical excisions and subsequent plastic procedures. In each case complementary radiotherapy was implemented. The metastases appeared in the lung 9, 17 and 21 years, respectively, after onset of the primary tumour as bilateral nodules in two cases and as a single nodule in one. Curative surgery was possible only in one subject, consisting in a bilateral metastasectomy via a midline sternotomy, with 5-year survival. Surgery was withheld in the other two subjects owing to advanced age and the presence of bilateral nodules with concomitant bone metastasis, respectively. Both were treated by chemotherapy and the patient with synchronous bone metastasis also received radiotherapy. The elderly subject died after six months, and the other patient after 19 months despite partial remission of disease. In our experience, curative therapy proves difficult due to multiple metastases. In the literature there are few reports regarding this rare type of metastatic lung disease, but surgical intervention would appear to be the therapy offering the best chance of long-term survival while chemotherapy constitutes the second-choice treatment. PMID- 10832543 TI - [The rationale of laparoscopic treatment in acute appendiceal disease]. AB - Notwithstanding its widely perceived advantages, laparoscopic appendectomy has not yet met with universal acceptance. The aim of the present work was to illustrate retrospectively the results of a case-control study of laparoscopic vs open appendectomy carried out at our institution. From Jan. 1993 to Dec. 1999 a total of 457 patients (M:F = 210:247; mean age 25.2 +/- 15 years) underwent emergency and/or urgent appendectomy. Among them, 254 (55.5%) were operated on laparoscopically, while 203 (44.5%) were treated by conventional surgery The choice of technique depended upon the availability or otherwise of a team expert in minimally invasive surgery. The laparoscopic technique conversion rate was 3.9% and was mainly due to the presence of dense intraabdominal adhesions. The major intraoperative complication rates were 0.39% and 0% in the laparoscopic and laparotomy groups, respectively (P = ns). Major postoperative complications occurred in 2 and 1%, respectively (P = ns). The postoperative mortality rates were 0.4% and 0.5% in the laparoscopy and laparotomy groups, respectively (P = ns). The reoperation rate was 1.1% in the laparoscopic group as against 0% in the open surgery group (P = ns). Minor postoperative complications were observed in 0.8% and 7.5% of patients in the laparoscopy and open surgery groups, respectively (P = 0.001) and consisted mainly of wound infections. Resumption of bowel function was significantly more rapid and the hospital stay significantly shorter in the laparoscopically treated patients. The greater diagnostic accuracy of laparoscopy allowed concurrent diseases to be diagnosed in 9% of laparoscopically treated patients with histologically proven appendicitis as against 1.5% of those treated by conventional surgery (P = 0.001). Similarly, among those patients with no evidence of gross and/or microscopic appendicitis, concurrent diseases were detected in 58.4% of the laparoscopic cases as against only 6% of the laparotomy cases (P = 0.0001). Despite the limitations of a retrospective investigation, on the basis of our experience we believe that laparoscopic appendectomy is as safe and effective as conventional surgery, presents a higher degree of diagnostic accuracy and makes for less trauma and a more rapid postoperative recovery. Such features make its use mandatory in female patients of child-bearing age referred for urgent abdominal and/or pelvic surgery. PMID- 10832544 TI - Secondary carcinoma of the liver from parotid gland tumor. AB - Secondary tumors of the liver from primary tumors arising in organs of the head and neck are rarely diagnosed during the patient's lifetime, though they should be suspected. A case of parotid mucoepidermoid carcinoma with liver metastases, treated by liver resection, is described. The clinical features and biological behavior of this secondary tumor are similar to those of the rare primary mucoepidermoid carcinoma of the liver. This case is unique because it is such a very rare occurrence. PMID- 10832545 TI - Spontaneous superior mesenteric vein thrombosis (SMVT) in primary protein S deficiency. A case report and review of the literature. AB - Superior mesenteric vein thrombosis (SMVT) is an uncommon but important clinical entity that can induce ischemia or infarction of the small and large bowel. It is rare and accounts for 5-15% of mesenteric vascular occlusions. Bowel infarction due to SMVT can present as an acute abdominal disease, requiring urgent laparotomy with resection of the intestinal segment affected. However, the clinical diagnosis of this event remains difficult and invariably requires specific imaging investigations in order to be able to treat the condition as soon as possible. SMVT without bowel infarction can present as persistent, non specific abdominal pain and nausea with minimal clinical signs, affecting young individuals without any known predisposing disorder, where laparotomy is not an urgent indication. We report a case of a young adult man with SMVT due to a hypercoagulable state (protein S deficiency), in whom an early diagnosis and appropriate anticoagulant treatment prevented any further extension of the thrombotic process and limited the hemorrhagic infarction of the ileum, which simply required a segmental resection. PMID- 10832546 TI - [Malignant biliary papillomatosis: a case report]. AB - The aim of this study was to describe the clinico-pathological characteristics of malignant biliary papillomatosis and the diagnostic and therapeutic approach adopted in this case, which extended from the left hepatic biliary duct to the bifurcation of the 2nd and 3rd segments. The patient was studied with ultrasonography, endoscopic retrograde cholangiopancreatography (ERCP), CT scan and MRI cholangiography, which showed the morphological characteristics and the extent of the disease in the biliary tract. The malignancy of the disease was confirmed by preoperative cytological analysis of bile collected via a nasobiliary probe. The surgical treatment, preceded by intraoperative ultrasonography, consisted in a left hepatectomy, sparing the 1st segment, and a right intrahepatic jejunostomy. The final pathological analysis revealed a malignant biliary papillomatosis. Preoperative ERCP clearly documented the exact distribution of the biliary tract lesions and allowed both treatment of the cholestasis and diagnosis of the nature of the lesion through cytological analysis of the bile. Surgical treatment must be carried out and guided not only by the preoperative diagnosis, but also by perioperative ultrasonography. Finally, owing to the rarity of such cases and consequently our very limited knowledge of the disease, an intensive follow-up is recommended. PMID- 10832547 TI - [Isolated rupture of aneurysm of the internal iliac artery]. AB - Isolated aneurysms of the internal iliac artery are rare and often asymptomatic. They are frequently diagnosed as a result of complications such as rupture. The authors describe in detail two cases of ruptured aneurysms of the internal iliac artery treated by emergency surgery with no perioperative mortality or morbidity. PMID- 10832548 TI - The development of normal fear: a century of research. AB - This paper reviews over a century's research into the developmental patterns of normal fear. Normal fear has been defined as a normal reaction to a real or imagined threat and is considered to be an integral and adaptive aspect of development with the primary function of promoting survival. Across a wide range of methodologies and assessment instruments researchers have been particularly focussed on investigating whether fear content, prevalence and intensity differ depending upon age, gender, socio-economic status, and culture. The structure and continuity of normal fears have also received much attention. The most consistently documented findings include that fear decreases in prevalence and intensity with age and that specific fears are transitory in nature. There are also predictable changes in the content of normal fear over the course of development. Such changes are characterized by a transition from infant fears which are related to immediate, concrete and prepotent stimuli, and which are largely non-cognitive, to fears of late childhood and adolescence which are related to anticipatory, abstract, and more global stimuli and events. Recent research into normal fear has more closely examined the validity of the more frequently used current assessment technique (i.e., the fear survey schedule). This research has provided some encouraging results as well as directions for future investigation. PMID- 10832549 TI - Issues in the diagnosis and assessment of anxiety disorders in children and adolescents. AB - Accurate diagnosis and assessment of anxiety disorders in children and adolescents is important for both treatment and research. In this paper, we review research related to the discriminant validity of the childhood anxiety disorders, and outline methods of assessment for children, including diagnostic interviews, self-report instruments, and behavioural, cognitive, and psychophysiological measures. Particular attention is given to psychometric and developmental issues, and their influence on assessment. The evidence provides support for the validity of the anxiety disorders as a whole, but only partial support for differentiation between specific anxiety disorders in children. Similarly, assessment methods used with anxious children provide reasonably accurate information on anxious symptoms, however are limited by poor discriminant validity and lack of sensitivity to developmental levels. PMID- 10832550 TI - Treatment of childhood anxiety: developmental aspects. AB - This review focuses on research on the cognitive-behavioral treatment of childhood anxiety disorders. Early forms of therapy for childhood anxiety were borrowed from adult treatment models. More recently, there has been a recognition of the need to design treatment from a child-based perspective. Consequently, several cognitive-behavioral programs designed specifically for children and youth have been both developed and evaluated. The importance of parental involvement has also been recognised in these treatment innovations. However, a number of developmental factors have yet to be given adequate consideration in both the research and practice of childhood anxiety treatment. The article highlights some of these factors including issues of individual, family and cultural variation. PMID- 10832551 TI - Research on the cognitive-behavioral treatment of school refusal: a review and recommendations. AB - Cognitive-behavior therapy is frequently used in the treatment of school refusal, a challenging problem for mental health professionals and school authorities. We review the clinical and research support for the efficacy of cognitive-behavior therapy using recently published guidelines for determining the level of evidentiary support for psychosocial interventions. Although cognitive-behavior therapy appears to be a useful treatment for school refusal, further research is needed before it can be considered as having "well-established" empirical status. Several other important methodological and theoretical issues are emphasized. PMID- 10832552 TI - Prevention of childhood anxiety disorders. AB - Anxiety disorders represent one of the most common and debilitating forms of psychopathology in children. While empirical research, mental health funding, and mental health professionals continue to focus on the treatment rather than prevention of anxiety disorders in children, preliminary research presents an optimistic picture for preventative strategies in the future. Knowledge of the risk factors, protective factors, and treatment strategies associated with childhood anxiety disorders, in conjunction with theories regarding the methods, timing, levels, and targets of prevention, equip us well for effectively preventing childhood anxiety disorders in the future. PMID- 10832553 TI - Quantification of the physiological relevance of a coronary stenosis using myocardial contrast echocardiography. AB - MCE can be used in the catheterization laboratory or in the operating room to provide rapid assessments of the functional significance of a coronary stenosis from direct arterial injections of microbubbles. In the past few years, the development of more stable microbubble contrast agents, and a better understanding of the interactions between ultrasound and microbubbles have led to the development of a truly non-invasive approach to quantify MBF using venous infusions. Furthermore, additional insights into the physiology of coronary stenosis, particularly as it affects MBV, have been obtained using MCE. PMID- 10832554 TI - Advances in microbubble technology. PMID- 10832555 TI - New imaging technology: measurement of myocardial perfusion by contrast echocardiography. AB - Myocardial perfusion imaging has long been a goal for the non-invasive echocardiographic assessment of the heart. However, many factors at play in perfusion imaging have made this goal elusive. Harmonic imaging and triggered imaging with newer contrast agents have made myocardial perfusion imaging potentially practical in the very near future. The application of indicator dilution theory to the coronary circulation and bubble contrast agents is fraught with complexities and sources of error. Therefore, quantification of myocardial perfusion by non-invasive echocardiographic imaging requires further investigation in order to make this technique clinically viable. PMID- 10832556 TI - Measurement of myocardial perfusion by contrast echocardiography: application in the operating room. PMID- 10832557 TI - The use of myocardial contrast echocardiography in clinical evaluation after myocardial infarction. AB - Because disruption of the microvasculature is a hallmark of myocyte necrosis, MCE may be able to distinguish between viable and infarcted tissue. In order to interpret images appropriately following myocardial infarction, however, one should be versed in the pathophysiology of post-ischemic reflow, and understand that reperfusion to infarcted tissue is a heterogeneous combination of hyperemia, low-reflow, no-reflow, and impaired microvascular flow reserve. Furthermore, the relative mix of these perfusion patterns changes both temporally and spatially, which has implications for the timing of MCE following reperfusion. The identification of no- and low-reflow by MCE predicts regions unlikely to demonstrate segmental functional recovery, and is associated with adverse clinical events. To date, studies documenting the utility of MCE in the AMI setting have been performed using intracoronary injections in the cardiac catheterization laboratory. With the advent of intravenous contrast agents and innovations in ultrasound imaging systems, it may be possible to make these determinations without the need for coronary instrumentation, thus expanding the role of MCE in acute infarction and reperfusion to settings such as the emergency room and intensive care unit. PMID- 10832558 TI - Myocardial perfusion by contrast echocardiography: diagnosis of coronary artery disease using contrast-enhanced stress echocardiography and assessment of coronary anatomy and flow reserve. AB - The advent of intravenous contrast agents, and newer ultrasound technology to enhance their detection, promises to improve and augment our conventional stress echocardiographic practice by improving diagnostic accuracy and providing novel information regarding myocardial perfusion and functional assessment of the coronary vasculature. The combination of intravenous contrast and harmonic stress echocardiography is a powerful tool for improved wall motion analysis through enhanced image quality, routinely permitting the evaluation of patients with suboptimal images. In this era of cost containment, we await studies in large populations addressing resource utilization and cost-effectiveness to determine if, indeed, all patients presenting with stress echocardiography should receive contrast. Myocardial perfusion can be observed using the technique, but the complex interactions of microbubbles and ultrasound in patients must be understood more fully before its implementation becomes routine practice. Non invasive imaging of coronary arteries using contrast-enhanced transthoracic harmonic echo/Doppler promises to expand the field of diagnostic and experimental echocardiography, bringing new insight into the pathophysiology of ischemic and non-ischemic heart disease. The continued development of newer contrast agents and refinement of ultrasound imaging equipment ensures that the applications of contrast echocardiography in the assessment of CAD will continue to increase. PMID- 10832559 TI - Long-term oestrogen treatment does not alter systemic arterial compliance and haemodynamics in biological males. AB - BACKGROUND: Studies have shown that chronic oestrogen treatment improves both lipid profile and vascular reactivity in postmenopausal women, in whom it also appears to have a beneficial effect on vascular haemodynamics and compliance. Whether oestrogen has a similar effect in men is unknown. OBJECTIVE: To determine whether long-term oestrogen treatment alters arterial compliance and haemodynamics in biological males. METHODS: We compared the effects of chronic oestrogen treatment on blood pressure, heart rate and arterial compliance in 21 male-to-female transsexuals prescribed long-term oestrogen treatment with those in 20 age-matched healthy males. Systemic arterial compliance was assessed using the 'area method', by the simultaneous measurement of aortic flow and driving pressure. RESULTS: Mean systemic arterial compliance was similar in transsexuals and age-matched males (mean +/- SE 0.66 +/- 0.06 ml/mmHg compared with 0.58 +/- 0.05 ml/mmHg, P = 0.34). These results did not differ after the exclusion of transsexuals with coronary risk factors or vascular disease. Heart rate (67 +/- 2 beats/min compared with 64 +/- 3 beats/min, P = 0.41), systolic blood pressure (119 +/- 3 mmHg compared with 119 +/- 2 mmHg, P = 0.95), pulse pressure (55 +/- 3 mmHg compared with 50 +/- 2 mmHg, P = 0.13), diastolic blood pressure (64 +/- 2 mmHg compared with 69 +/- 2 mmHg, P = 0.06) and mean arterial pressure (84 +/- 2 mmHg compared with 89 +/- 2 mmHg, P = 0.09) were also similar at baseline between the two groups. Serum testosterone (an index of oestrogen treatment) was markedly suppressed in the transsexuals compared with the males (0.8 +/- 0.5 nmol/l compared with 25.3 +/- 12.6 nmol/l, P < 0.0001). Univariate analysis revealed that the best predictors of arterial compliance were the pulse pressure (rs = 0.41, P = 0.02) and the systolic blood pressure (rs = -0.35, P = 0.02). On multivariate analysis, the best combination of predictors of compliance were the pulse pressure, testosterone and low-density lipoprotein cholesterol concentrations (R2 = 0.29, P = 0.01). CONCLUSIONS: Although previous evidence suggests chronic oestrogen treatment can improve endothelium-dependent vasodilatation and favourably alter the lipid profile in biological males, these changes are not reflected in changes in systemic arterial compliance. Changes in arterial compliance may not be central to the beneficial effects of oestrogen on vascular function, at least in males. PMID- 10832560 TI - Cardiac rotation and relaxation after anterolateral myocardial infarction. AB - BACKGROUND: Both systolic and diastolic dysfunction have been observed in patients with anterolateral myocardial infarction. Diastolic dysfunction is related to disturbances in relaxation and diastolic filling. OBJECTIVE: To analyse cardiac rotation, regional shortening and diastolic relaxation in patients with anterolateral infarction. METHODS: Cardiac rotation and relaxation in controls and patients with chronic anterolateral infarction were assessed by myocardial tagging. Myocardial tagging is based on magnetic resonance imaging and allows us to label specific myocardial regions for imaging cardiac motion (rotation, translation and radial displacement). A rectangular grid was placed on the myocardium (basal, equatorial and apical short-axis plane) of each of 18 patients with chronic anterolateral infarction and 13 controls. Cardiac rotation, change in area and shortening of circumference were determined in each case. RESULTS: The left ventricle in controls performs a systolic wringing motion with a clockwise rotation at the base and a counterclockwise rotation at the apex when viewed from the apex. During relaxation a rotational motion in the opposite direction (namely untwisting) can be observed. In patients with anterolateral infarction, there is less systolic rotation at the apex and diastolic untwisting is delayed and prolonged in comparison with controls. In the presence of a left ventricular aneurysm (n = 4) apical rotation is completely lost. There is less shortening of circumference in infarcted and remote regions. CONCLUSIONS: The wringing motion of the myocardium might be an important mechanism involved in maintaining normal cardiac function with minimal expenditure of energy. This mechanism no longer operates in patients with left ventricular aneurysms and operates significantly less than normal in those with anterolateral hypokinaesia. Diastolic untwisting is significantly delayed and prolonged in patients with anterolateral infarction, which could explain the occurrence of diastolic dysfunction in these patients. PMID- 10832561 TI - Predictors of left ventricular thrombus formation in patients with anterior myocardial infarction: role of activated protein C resistance. AB - BACKGROUND: Left ventricular mural thrombus formation is a well-recognised consequence of acute anterior myocardial infarction. The vast majority of left ventricular thromboses occur in patients with anterior myocardial infarction and depressed left ventricular function. OBJECTIVE: To evaluate the factors predicting left ventricular thrombus formation in patients similar for left ventricular function and left ventricular score indexes. METHODS: We evaluated 45 consecutive patients who met the inclusion criteria of anterior myocardial infarction resulting in apical, anterior or septal asynergy (akinesia, dyskinesia), without non-Q-wave myocardial infarction, dilated cardiomyopathy, or renal or hepatic dysfunction. Patients were divided into two groups: group I with, and group II without, left ventricular mural thrombus. The groups were compared for clinical, echocardiographic and hematologic parameters (activated protein C resistance (APC-R), protein S and antithrombin III). RESULTS: Smoking and ACP-R were significantly greater in group I than in group II (P < 0.05 and P < 0.005 respectively). Multivariate regression analysis showed that APC-R was an independent risk factor for left ventricular thrombus formation in the patient group selected. Antithrombin III and protein S concentrations were not statistically different between two groups. All other clinical and echocardiographic characteristics of the patients were similar in both groups. CONCLUSION: APC-R is an independent risk factor for left ventricular thrombosis in patients with anterior myocardial infarction resulting in septal or anterior and apical akinesia or dyskinesia. PMID- 10832562 TI - Histopathologic analysis of in-stent neointimal regression in a porcine coronary model. AB - BACKGROUND: Animal and clinical studies have demonstrated late regression of in stent neointima. This study was performed to identify the temporal changes in the in-stent neointimal constituents responsible for late regression. METHODS: NIR stents were implanted in porcine coronary arteries (size of stent (in mm) to size of artery (in mm) approximately equal to 1.1) and harvested at 2 months and 6 months (n = 6 stents/group). Histopathologic analyses included morphometric analysis, smooth muscle cell density, and extracellular matrix contents. RESULTS: Compared with the findings at 2 months, at 6 months there was a significant reduction in area stenosed (from 21 +/- 3% to 14 +/- 1%, P < 0.05) and neointimal thickness (from 0.2 +/- 0.03 mm to 0.03 +/- 0.02 mm, P < 0.05), despite similar injury scores (0.05 +/- 0.06 at 2 months and 0.36 +/- 0.29 at 6 months). This regression was accompanied mainly by a reduction in proteoglycan (from 24 +/- 19% to 5 +/- 8%, P = 0.05), with no change in smooth muscle cell density (71 +/- 7 compared with 76 +/- 23/high power field) or collagen content (25 +/- 19% compared with 25 +/- 19%). CONCLUSIONS: The study confirmed the regression of in stent neointima, which was mainly attributable to a reduction in proteoglycan content, resembling the natural healing response. PMID- 10832564 TI - Bibliography. Current world literature. PMID- 10832563 TI - Fructose-2,6-bisphosphate, a potent stimulator of phosphofructokinase, is increased by high exogenous glucose perfusion. AB - BACKGROUND: We have previously demonstrated that perfusion of isolated hearts with high concentrations of glucose results in increased glycolysis during ischemia, diminished ischemic injury, and improved functional recovery with reperfusion. OBJECTIVE: To evaluate a possible mechanism by which glucose conferred this protection. We examined the hypothesis that increased exogenous glucose concentrations results in increased concentrations of fructose-2,6 bisphosphate, a potent activator of phosphofructokinase-1, and thus increases glycolysis. METHODS: Perfused rabbit hearts were subjected to 60 min of low-flow ischemia. Control hearts were perfused with buffer containing 0.4 mmol/l palmitate, 5 mmol/l glucose, and 70 mU/l insulin, and treated hearts were perfused with buffer containing 0.4 mmol/l palmitate, 15 mmol/l glucose and 210 mU/l insulin. RESULTS: Ischemic contracture was attenuated by perfusion of high concentrations of glucose (high glucose) (P < 0.05 compared with control). Glucose uptake and lactate production were greater in hearts perfused with high glucose, as was the ATP concentration at the end of ischemia (P < 0.05 compared with controls). Exogenous glucose uptake and lactate production correlated well with fructose-2,6-bisphosphate content (P = 0.007). CONCLUSIONS: Enhancement of glycolysis in hearts perfused with high glucose may be the result of stimulation of phosphofructokinase-1 by fructose-2,6-bisphosphate. Accordingly, this may serve as an important mechanism by which cardioprotection may be achieved. PMID- 10832565 TI - Acculturation and health in Korean Americans. AB - Despite increasing research on the relationships between acculturation and health, it is unclear whether (1) ethnic group variation occurs in acculturation health relationships, (2) acculturation components vary differently in relationship to health, (3) biculturalism has beneficial effects on health and (4) multidimensional health relationships occur with acculturation. This study examined the Korean American ethnic group, considering how acculturation was related with five dimensions of health: smoking, physical activity, fat intake, body weight, and reported health. Pretested questionnaires were mailed to a national sample with Korean American surnames, and 55% of the deliverable sample responded, producing 356 usable questionnaires. Acculturation was measured using a two-culture matrix model and Gordon's theoretical work, and showed three distinct groups (acculturated, bicultural and traditional) and four components (American structural, American cultural, Korean structural and Korean cultural). Bicultural men were least likely to smoke, while acculturated and bicultural women were more likely to smoke than traditional women. Korean structural and cultural components were related to men's smoking. Higher acculturation was related to light physical activity, but not to vigorous physical activity. Fat intake did not differ by acculturation status. Higher acculturation was associated with higher body weight and better self-reported health only in men. Higher American cultural component scores were associated with better self reported health in men. Among Korean Americans, acculturation components varied in their relationships with health, beneficial effects of being bicultural on health were not found, and acculturation-health relationships were multidimensional. Overall, ethnic group variation in health occurred, with Korean Americans similar to some ethnic groups but different than others. Future health research and practice can benefit by acknowledging the complexity of acculturation and its multidimensional effects on health. PMID- 10832566 TI - Pain response in Chinese and non-Chinese Canadian infants: is there a difference? AB - This study was designed to compare the behavioural pain responses of 2-month-old Canadian-born Chinese babies receiving a routine immunization to those of non Chinese infants in similar situations. Two groups of 26 infants were obtained from a pediatric clinic held by a Chinese pediatrician and a suburban pediatric practice of a large Canadian city. Facial expression using the Neonatal Facial Coding System (Grunau, R.V.E., Craig, K.D., 1987. Pain expression in neonates: facial action and cry. Pain, 28, 395-410.) and cry using the Fast Fourier Transform were measured during 30 s following the insertion of the needle. Acculturation in Chinese mothers, circadian rhythm, gender, height and weight in infants were assessed. Multivariate analysis of variance revealed significant differences in pain response between these two groups with the Chinese babies showing greater response. No significant effect of circadian rhythm and gender was identified. These results suggest the presence of differences in acute pain response in relation to culture by at least 2 months of age. PMID- 10832567 TI - Indices and sociodemographic determinants of childhood mortality in rural Upper Egypt. AB - There is a general agreement that childhood mortality in Egypt has substantially declined during the last two decades. The decline has been traditionally attributed to the impact of the National Control of Diarrhoeal Diseases Program (NCDDP). However, a controversy still exists over the magnitude of that decline and the claimed impact of the NCDDP on diarrhoea related mortality. This study was carried out in six sites in rural Upper Egypt to determine indices, leading causes, and sociodemographic determinants of childhood mortality. Verbal autopsy was conducted with mothers or caretakers who had reported the death of a child under the age of five (U5) before the study to determine the leading cause of death. Then, the association between childhood mortality and a wide set of sociodemographic risk factors was examined by comparing these children with 1025 living U5 children using a multivariate logistic regression analysis. Results showed that the average infant and U5 mortality rates are 97.2 and 130.8 per 1000 live births respectively. Verbal autopsy revealed that the leading causes of U5 mortality are: diarrhoeal diseases (39.4%), acute respiratory infection (26.8%), combined episode of both (5.1%), febrile illnesses including meningitis (10.6%), neonatal causes (12.6%), and accidents (2.5%). Diagnosis was not determined in 3.0% of the cases. Child age (< 12 month) and mother age at childbirth are the strongest determinants of childhood mortality. Other determinants include parental illiteracy, parental age difference, house ownership, child order and average household meat consumption. PMID- 10832568 TI - The determinants of infant mortality in Pakistan. AB - This study examines factors associated with infant survival in Pakistan. It uses data from the Pakistan Integrated Household Survey 1991, a nationally representative sample survey of the Government of Pakistan, funded by the World Bank. The infant mortality rate was still very high in Pakistan until the early 1990s, at 100 deaths per 1000 live births. The study shows that there is no evidence of a secular decline in infant mortality during the 1980s. Large differentials in infant survival by socio-economic factors and access to water and sanitation indicate that social and gender inequities are the underlying cause of the stagnation of infant mortality in Pakistan. Economic and social policies of earlier decades have resulted in tremendous disparities in wealth and access to resources in Pakistan. The low social, economic and legal status of women is intimately tied to the well-being of their children. Health interventions in Pakistan should be designed to reach the most under-served: women and children. Systematic evaluations of health interventions will be necessary to make informed decisions about health investments in the future. PMID- 10832569 TI - Talking contracts and taking care: managers and professionals in the British National Health Service internal market. AB - This paper draws on audio-recordings of a Health Authority's contract monitoring meetings with hospital providers to examine the nature of interactions between managers and clinical professionals in the British National Health Service internal market. It describes how managers and professionals arrive at a working division of labour, which acknowledges their respective spheres of expertise, but also leaves a space where definitions are contested. There is an interplay of competing managerial and professional discourses which construct problems and proposed solutions in fundamentally non-commensurable ways. Yet only rarely do managers or professionals mount challenges in ways that bring them on to the territory of the other group; rather each group seeks to frame problems to mesh with its special domain of competence. Managers seek to push back the boundaries of professional control by constructing a language and set of practices which will govern the contracting process. But they remain reliant on professionals to mediate between the requirements of contracting and the realities of clinical work. Clinical arguments continue to have high perceived legitimacy for managers, and will often be taken up by hospital managers in negotiations with their Health Authority counterparts. It is argued that the dependency of managers on professionals to make the contracting system work, taken together with the continued social and cultural authority of senior medical consultants, limits managers' ability to control professionals. PMID- 10832570 TI - Cost analysis as a vitamin A program design and evaluation tool: a case study of the Philippines. AB - Vitamin A deficiency (VAD) is a serious and widespread public health problem in the Philippines. Initiated in 1993, the Philippines National Vitamin A Supplementation Program (NVASP) is one of the oldest, most mature and comprehensive of its kind. This paper presents a cost-effectiveness and efficiency analysis of the NVASP and of a hypothetical program of vitamin A fortification of wheat flour that was conducted to inform policymakers as to how to modify the program. Employing a proxy effectiveness indicator of VAD--the intake of < 70% of the recommended daily allowance of vitamin A--in a series of simulations using individual child consumption data, the analysis finds that fortification is more efficient in reducing inadequate vitamin A intake (IVAI) compared to the NVASP. Due to the nature of food consumption patterns, however, fortification alone, is not enough. At what is regarded as the maximum politically acceptable fortification level, there will still be 2.2 million (29%) Filipino children aged 12-59 months who will have IVAI. An investigation of the cost and efficiency of geographically targeted supplementation programs reveals that maintaining a universal supplementation program in urban areas and, in rural areas, introducing a targeted program to only the poorest municipalities (where the prevalence of VAD is the highest) will provide a more acceptable public health policy response than fortification alone. Such a policy will reduce incremental direct Government expenditures on vitamin A programs by nearly 20% and will reduce the number of children with IVAI to 900,000 (12%) Filipino children. The paper describes the fortification and supplementation programs, and how their costs were estimated. Lessons for program designers and policymakers in other countries in which vitamin A deficiency constitutes a public health problem are also discussed. PMID- 10832571 TI - Precision and accuracy in measuring absence from work as a basis for calculating productivity costs in The Netherlands. AB - The impact of disease on the ability of a person to perform work should be part of an economic evaluation when a societal viewpoint is used for the analysis. This impact is reflected by calculating productivity costs. Measurement of these costs is often performed retrospectively. The purpose of our study was to study precision and accuracy of a retrospective self-administered questionnaire on sick leave. Employees of a company were asked to indicate the number of days absent from work due to illness during the past 2 weeks, 4 weeks, 2 months, 6 months, and the past 12 months. The percentage of respondents with an absolute difference of a maximum of respectively 0, 1, 2, 3, 4, 5, 6, 7, 8, and 9 or more days between reported, and company-registered absence due to illness was determined. Besides this, the proportional difference was calculated. A systematic difference was tested with a signed rank test. Of the reported data, 95% matched the registered data perfectly when the recall period was limited to 2 and 4 weeks. This percentage decreased to 87%, 57%, and 51% for 2 months, 6 months, and 12 months. The weighted mean proportional differences for the recall periods were respectively 32.9, 35.2, 45.3, 34.9, and 113.6%. No systematic positive or negative difference was found between registered and reported sick leave. The results suggest that the recall period for retrospective measurement of sick leave is limited according to the precision level, which seems to be appropriate for the subject and the purpose of the study. We recommend using a recall period of no more than 2 months. PMID- 10832572 TI - Lay constructions of HIV and complementary therapy use. AB - This study examines the meanings that individuals with HIV attach to their use of complementary therapies. A qualitative analysis of 66 interviews completed between 1993 and 1998 showed that complementary therapies represent different things for these individuals--a health maintenance strategy, a healing strategy, an alternative to Western medicine, a way of mitigating the side-effects of drug therapies, a strategy for maximizing quality of life, a coping strategy, and a form of political resistance. We found that the meanings individuals ascribe to complementary therapies and the benefits they expect to derive from them are not idiosyncratic, but linked to social characteristics--sexuality, ethnocultural background, gender--and to beliefs about health and illness, values and experiences. We found as well that these meanings are neither mutually exclusive nor fixed. The therapies often appeal to individuals on different levels and their appeal may change over time. PMID- 10832573 TI - Walking, exercising, and smoking: does neighborhood matter? AB - Neighborhood context could affect health behaviors because of structure or contagion. We expected that residents of US neighborhoods where a high percentage of residents are poor and do not have college degrees would be more likely to smoke and less likely to walk and exercise. We examined the hypotheses using multi-level data in which survey information from a representative sample of Illinois residents is linked to census-tract information about poverty and education in their neighborhood. Contrary to expectations we found that residents of poor neighborhoods were more likely to walk than those in less disadvantaged places, adjusting for individual poverty, household income, education, race, ethnicity, sex, age, and marital status. This was the case despite the fact that residents of poor neighborhoods were more afraid to leave the house and feared being victimized on the streets. Consistent with expectations we found that residents of neighborhoods where a high percentage of residents are college educated are more likely to walk. Thus, the two aspects of neighborhood socioeconomic status had opposite effects on walking. Neighborhood context had no effect on the likelihood of exercising strenuously. Men in poor neighborhoods were more likely to smoke than those in less disadvantaged places, but neighborhood context had no significant effect on women's likelihood of smoking. PMID- 10832574 TI - Gender, poverty and location: how much difference do they make in the geography of health inequalities? AB - It is often said that women live longer than men, but suffer more illnesses throughout their lives. It has also been demonstrated in various studies of women's health that measures of health and health behaviour vary over different geographic scales. Added into this mix is the fact that historically more women than men in relative terms are found on the lower rungs of the socio-economic ladder. What has not been so well-developed is our understanding of the connections among health, gender, poverty and especially location. In 1998, Statistics Canada released the second wave of the National Population Health Survey (NPHS-2). Included with the NPHS-2 public use microdata file are measures of health status, gender, income and location which can be analyzed in the form of logistic regression models. Results are reported which provide a better understanding of the relative roles that gender, poverty and location play in the geography of inequalities. PMID- 10832575 TI - Urban-rural mortality differentials: controlling for material deprivation. AB - This paper investigates the relationship between premature mortality and material deprivation, and the differences in this relationship between urban and rural areas. We examine, given comparable measures of affluence or deprivation, whether residual differences exist between urban and rural areas for all-causes of death and, separately, for cancers, circulatory and respiratory diseases. Using 1990-92 mortality data for the 908 wards of Wales we apply statistical analyses based on tabular data and parametric Poisson regression models. Contrasts are sought between six urban and rural categories defined in terms of settlement sizes and the employment structure of rural areas. Inequalities in all-cause premature mortality are widest in the cities, narrowest in the deeper rural areas, and of intermediate and comparable value in other areas of Wales. This is largely a reflection of the different distributions of material deprivation in these areas. After controlling for differences in socio-economic characteristics, using deprivation measures, the tendency for lower mortality in deeper rural areas is substantially reduced. Residual mortality differences between urban and rural areas are shown to be dependent on the way deprivation is measured and the disease group under study. For cancers there are no residual mortality differences, while for respiratory and circulatory diseases some of the residual variation can be accounted for by employment variables, particularly previous employment in the coal mining industry. PMID- 10832576 TI - Heterogeneity in the determinants of health and illness: the example of socioeconomic status and smoking. AB - Systematic variations in health and illness among social groups have persisted and, in some cases increased, in many countries in spite of improvements in the availability of, and access to, health care services. Health policy makers have responded by showing increasing interest in non-clinical determinants of health as a way of explaining the observed systematic variations in health and illness. Yet health care and non-health care "factors" are often seen as competing for society's scarce resources in the production of health. The purpose of this paper is to augment this traditional approach to understanding the determinants of health in populations by exploring heterogeneity in the smoking-health relationship between social groups. Logistic regression analyses were performed using data from the 1992-93 Sante Quebec survey, a weighted random sample of the non-institutionalized population of the province of Quebec, Canada (N = 23,564). The findings suggest that the likelihood of reporting health as poor or fair was greater for smokers than non-smokers. However, the difference between smokers and non-smokers was significantly greater among groups of the population with low incomes and without employment, but significantly less among sub-groups with lower levels of education. These findings suggest that the identification, evaluation and implementation of health care programmes need to pay greater attention to the interface between social, behavioural and clinical determinants of health. PMID- 10832577 TI - Congenital disorders of glycosylation (CDG): a rapidly expanding group of neurometabolic disorders. PMID- 10832578 TI - Congenital disorder of glycosylation-Ic: case report and genetic defect. AB - The clinical phenotype and the molecular defect of a patient with a new subtype of congenital disorders of glycosylation (CDG-Ic, formerly designated as CDGS type V) characterized by a deficiency of Dol-P-Glc: Man9GlcNAc2-PP-Dol glucosyltransferase is described. The clinical picture presents with several features similar to CDG-Ia (phosphomannomutase 2 deficiency) such as hypotonia and atactic-dystonic movements. In contrast to CDG-Ia, the course of the disease appears milder. The head growth, the functioning of the peripheral nerves and the initial cerebellar development were normal. Sequencing of the patient's Dol-P Glc: Man9GlcNAc2-PP-Dol glucosyltransferase cDNA revealed an in-frame deletion of three nucleotides leading to the loss of isoleucine 299. PMID- 10832579 TI - Reorganization of the hand somatosensory cortex following perinatal unilateral brain injury. AB - Functional magnetic resonance imaging was used to map the hand somatosensory cortices of nine hemiparetic young adult patients with perinatal unilateral brain injury in the sensorimotor area and five normal subjects. Stimulation of the paretic hand by periodic manual squeezing produced activation in the contralateral hemisphere of three patients and in the ipsilateral hemisphere of three other patients. Paretic hand stimulation produced no activation in either hemisphere of the remaining three patients. Therefore, one-third of the patients demonstrated functional "plasticity" of the brain in the form of inter hemispheric relocation of the hand somatosensory function. The volume and pattern of activation for both hands was altered for those patients that showed evidence of cortical reorganization to the opposite hemisphere. This differs from the hand motor system, which exhibited inter-hemispheric reorganization in a higher proportion of a related group of hemiparetic subjects. PMID- 10832580 TI - Unilateral involuntary movement associated with streptococcal infection: neurophysiological investigation. AB - Two boys developed rhythmic involuntary movements in the extremities on one side of the body after febrile illness. They also showed behavioral disturbances. In both patients, serum antistreptolysin-O and antistreptokinase titers were elevated in acute illness and decreased a few months later. One patient showed tremorous movement, and the other choreiform movement. In the former, a surface EMG showed short-duration (30 to 60 ms), highly frequent (6 to 8 Hz) and synchronous discharges of multiple muscles, including the antagonists, suggesting myoclonic jerk. In the latter, a surface EMG showed long-duration (0.5 to 1 s), repetitive (about 0.5 Hz) and synchronous or asynchronous discharges of the antagonists, suggesting choreoathetosis. In both patients, giant somatosensory evoked potentials and high-voltage slow EEG activities were observed predominantly in the hemisphere contralateral to the involuntary movement. In the myoclonic patient, long-latency EMG responses were enhanced and cortical potentials preceding the myoclonus were present by jerk-locked back averaging technique. The present data suggest that unilateral rhythmic involuntary movements occur secondary to streptococcal infection. The pathophysiology of the involuntary movements may be associated with sensorimotor cortex hyperexcitability. PMID- 10832581 TI - Changes in cerebral blood volume and cerebral oxygenation during periodic breathing in term infants. AB - The present study is an analysis of changes in cerebral oxygenation and cerebral blood volume (CBV) during periodic breathing in healthy term infants by means of near infrared spectroscopy (NIRS). Polygraphy included electrocardiogram, electrooculogram, heart rate, oxygen saturation, side stream capnography, two respiratory effort sensors, a movement sensor, and NIRS. During periodic breathing analysis of behaviour of total haemoglobin (cHbtot), deltaCBV, the haemoglobin oxygenation index (cHbD), and cytochrome oxidase (CytOx) was performed. In ten healthy term infants 30 cycles of periodic breathing with a mean of 10 apnoeas per cycle were analysed. Corresponding cyclical variations of cHbD appeared in 98%, cyclical variations of cHbtot appeared in 42% of all apnoeas. During phases of apnoea, a mean decrease of cHbD of -3.45 micromol/l occurred 1.75 seconds after onset of apnoea, and a mean decrease of cHbtot of 0.79 micromol/occurred 0.74 seconds after onset of apnoea. During these apnoeas, the deltaCBV was -44 microl/100 g brain. During phases of ventilation, there was an increase of cHbD and cHbtot to the pre-apnoeic levels. There was a tendency that CytOx values decreased during periodic breathing, the amount of decrease was -0.32 micromol/l. In conclusion, the present study was able to show for the first time that there is cyclical desaturation and reoxygenation of cerebral blood during periodic breathing. Cyclical changes in CBV in association with periodic apnoea occurred only in 42% of apnoea. PMID- 10832582 TI - Inheritance of photosensitivity. AB - PURPOSE: To investigate the mode of inheritance of the photoparoxysmal response (PPR) and to obtain more information about the influence of photosensitivity on the seizure risk in siblings of patients with epilepsy. METHODS: Examination of the records of families with one photosensitive parent (Group I, n = 54) and of families with a photosensitive proband, neither of whose parents was photosensitive (Group II, n = 72). RESULTS: At the age of maximum penetrance, between 5 and 15 years of age, 50% of the siblings in Group I were photosensitive, compared to only 15% in Group II. Siblings in Group I showed a higher seizure rate (19%) than siblings in Group II (4%, p < 0.005). The majority of photosensitive siblings had no seizures, but photosensitive siblings had a higher seizure risk than non-photosensitive siblings. The highest seizure risk was found in photosensitive siblings of Group I (33%) compared to 9% in non photosensitive siblings (p < 0.05). CONCLUSION: Our findings show that a PPR in parents is a major determinant for the risk of a PPR in offspring. The results may indicate an autosomal-dominant transmission with age-dependent penetrance of the PPR. Photosensitivity is an important factor in the pathogenesis of seizure disorders in childhood. PMID- 10832583 TI - Alexander disease--classification revisited and isolation of a neonatal form. AB - Alexander disease is usually classified according to the age of onset, e.g. an infantile form with onset during the first two years of life, a juvenile form with onset in childhood, mainly school age. It has been recognized, however, that the clinical course can be very variable within these groups. Thus, this clinical classification is not a useful predictor of severity and progression of the disease. This is demonstrated here on the basis of the history of seven own patients and a literature review. Only an onset in very early infancy, during the neonatal period, seemed to be associated with a rather uniform pattern of disease course, often leading to early death. This neonatal form showed very stereotyped symptoms, in part different from later onset: Early, often intractable, generalized seizures; hydrocephalus with raised intracranial pressure due to aqueductal stenosis because of pathological astroglia proliferation; lack of developmental progression but without prominent spasticity or ataxia; elevated CSF protein content. This was associated with the well-established neuroradiological findings, e.g. severe white matter affection with fronto temporal predominance, involvement of basal ganglia and periventricular enhancement as an obligatory symptom. The identification of this early onset form is especially important as seizures and signs of raised intracranial pressure may mislead the diagnosis. PMID- 10832584 TI - Novel cluster of tRNALeu(UUR) mutations in a sporadic case of infantile myopathy restricted to muscle tissue. AB - In a previous study we reported on a case with severe infantile, mitochondrial myopathy caused by somatic mutation [12]. In the present study we give evidence for asymmetric tissue distribution of the mutations. Mitochondrial DNA (mtDNA) analysis showed a cluster of nearly homoplasmic point mutations in the tRNA gene for leucine (UUR) (A3259 G, A3261 G, A3266 G, A3268 G). The mutation is abundant in muscle, but is not found in blood cells. This cluster of mutations is sporadic, because the search for mutant molecules in the blood of the healthy mother and maternal grandmother did not show these alterations. PMID- 10832585 TI - Cerebellar involvement as a rare complication of pneumococcal meningitis. AB - A 4-year old girl with meningitis, caused by streptococcus pneumoniae, developed a subcoma with respiratory insufficiency, followed by a severe cerebellar syndrome. Cerebellar involvement after regaining consciousness consisted of a symmetrical ataxia and mutism. This mutism changed into dysarthria and finally into normal speech. Magnetic resonance imaging revealed lesions in both cerebellar hemispheres, suggesting cerebellitis. She recovered with prompt antibiotic treatment. PMID- 10832586 TI - A case of leukoencephalopathy with vanishing white matter. AB - A 6-year old Turkish boy with a recently defined entity: "leukoencephalopathy with vanishing white matter" is described. He was born to consanguinous parents. His psychomotor development was normal till he first presented with fever and generalized tonic-clonic seizures at the age of 2.5, followed by rapid motor and mental deterioration. Decerebrate posture and marked spasticity subsequently developed. The initial MRI examination showed diffuse involvement of white matter, including subcortical U-fibers, with signal intensity parallel to CSF on all sequences. The white matter appeared swollen. The ventricles were slightly enlarged and there was cavum septi pellucidi et vergae. The posterior crus of the internal capsule, external and extreme capsules were affected. Cerebellar hemispheres and vermis showed atrophy. The involvement pattern of brainstem was noteworthy in that pontine tegmentum and cruri cerebri were affected. Follow-up MRI obtained after three years did not show any interval change. Brain biopsy showed thinned cortex with relatively preserved cortical layering and neuronal structure. There was rarefaction of the white matter with cystic degeneration. Fibrillary gliosis and increased number of oligodendroglial cells were observed within the cerebral white matter. PMID- 10832587 TI - Fetal-onset severe skeletal muscle glycogenosis associated with phosphorylase-b kinase deficiency. AB - We report on a premature newborn girl delivered after 32 weeks of gestation by cesarean section after sparse limb movements, fetal tachycardia and late heart rate decelerations had suggested fetal distress. Following 1 day of mechanical ventilation, adequate pulmonary gas exchange was achieved by spontaneous breathing. Main symptoms were virtually complete absence of spontaneous movements, increased flexor tonus of the extremities, and hypotonia of the trunk. Inability to suck or swallow required nasogastric gavage feeding. There were no hypoglycemic episodes. Echocardiography revealed normal myocardial function. Creatine kinase was 237 U/I at 2 days of life, declining to normal values thereafter. Muscle biopsy revealed increased glycogen storage with subsarcolemmal glycogen deposits and low phosphorylase-a activity while total phosphorylase was normal after in vitro activation, suggestive of phosphorylase-b kinase deficiency. No mutation was detected in exon 1 of the myophosphorylase gene. No psychomotor development was observed, and the infant died of central apnea at 3 months of age. PMID- 10832588 TI - Chorea as the presenting clinical feature of primary antiphospholipid syndrome in childhood. PMID- 10832589 TI - Atypical features in Angelman syndrome due to imprinting defect of uniparental disomy of chromosome 15. PMID- 10832590 TI - Effect of social housing condition on heat shock protein (HSP) expression in the Shionogi mouse mammary carcinoma (SC115). AB - Our previous studies have shown that social housing conditions can significantly alter the growth rate of the Shionogi mouse mammary carcinoma (SC115). The present study extended our investigations to the molecular level by examining stressor effects on the expression of a group of stress-responsive proteins, the heat shock proteins (HSPs). We hypothesized that HSP expression in SC115 cells may be altered by (a) different social housing conditions in vivo and (b) steroid hormone and growth factor exposure in vitro. Mice were reared in groups (G) or as individuals (I). Immediately following tumor cell injection, mice were rehoused from group to individual (GI), from individual to group (IG), or they remained in groups (GG). Tumor tissue was resected at 0.8 g or 3.0 g, as evidence suggests that tumor size affects HSP expression, which in turn affects proliferation. The data demonstrate that expression of HSP25, 70, and 90 was increased in tumors from mice in the IG compared to GG and GI mice, at both tumor weights examined. In addition, in IG mice, HSP90 expression was greater in 0.8 g compared to 3.0 g tumors. Under controlled culture conditions, hormones known to stimulate SC115 growth both in vivo and in vitro altered HSP expression. Physiological levels of dihydrotestosterone (DHT) and pharmacological levels of hydrocortisone (HC) upregulated expression of HSP25, whereas physiological levels of beta-estradiol (E2) upregulated expression of HSP90. These data are the first to demonstrate that a psychosocial stressor, a change in social housing condition, can induce differential HSP expression. Further, these data show that hormones that regulate SC115 tumor growth, also alter HSP expression. PMID- 10832591 TI - Quantitative measurement of soluble cytokeratin fragments in tissue cytosol of 599 node negative breast cancer patients: a prognostic marker possibly associated with apoptosis. AB - Apoptosis is associated with caspase-mediated proteolysis of Type I (K18 and K19) cytokeratins. We previously showed a positive association between the levels of tissue polypeptide antigen (TPA), that recognizes cytokeratins K8, K18, and K19 fragments, and induced apoptosis in breast cancer cell lines. The aim of the present study was to evaluate the interrelationships between TPA, steroid receptors, and p53, and their joint prognostic role in node-negative breast cancer patients not treated with adjuvant therapies. Age and pT were also considered since they are known prognostic factors. Five hundred and ninety-nine cases with N- breast cancer were evaluated (median follow-up: 60 months). TPA was measured by an immunoradiometric assay and p53 by an immunochemiluminescent assay in tumor cytosol. Multiple correspondence analysis was used to study the associations among variables. Their prognostic role (univariate analysis) and their joint effect (multivariate analysis) on RFS were investigated with Cox regression models. TPA showed a direct association with ER and PgR. Higher p53 values were weakly associated to low values of ER, PgR, and TPA. Younger age was related to low and intermediate values of ER and PgR and to low p53 values, while older age was related to high values of ER. Multivariate analysis showed a significant prognostic impact for pT, age, ER, and TPA. Among the interactions considered clinically relevant, only that between ER and age was found. RFS estimated values were poorer in cases with lower than in those with higher TPA values, both in patients expected to have a poor (pT2, young age, low ER) and a better prognosis (pT1, older age, high ER). From the findings of the present study we can draw the following conclusions: The relationship of TPA with prognosis gives an additional contribution to pT, age, and steroid receptors in N breast cancer; TPA may be considered the first marker of apoptosis measured with a fully standardized quantitative method in tumor cytosol and could be evaluated in prognostic indexes including markers related to different biological mechanisms. PMID- 10832593 TI - Role of specific apoptotic pathways in the restoration of paclitaxel-induced apoptosis by valspodar in doxorubicin-resistant MCF-7 breast cancer cells. AB - Paclitaxel (Taxol) kills tumor cells by inducing both cellular necrosis and apoptosis. A major impediment to paclitaxel cytotoxicity is the establishment of multidrug resistance whereby exposure to one chemotherapeutic agent results in cross-resistance to a wide variety of other drugs. For example, selection of MCF 7 breast cancer cells for resistance to doxorubicin (MCF-7ADR cells) results in cross-resistance to paclitaxel. This appears to involve the overexpression of the drug transporter P-glycoprotein which can efflux both drugs from tumor cells. However, MCF-7ADR cells possess a deletion mutation in p53 and have considerably reduced levels of the Fas receptor, Fas ligand, caspase-2, caspase-6, and caspase 8, suggesting that paclitaxel resistance may also stem from a bona fide block in paclitaxel-induced apoptosis in these cells. To address this issue, we examined the ability of the P-glycoprotein inhibitor valspodar to restore paclitaxel accumulation, paclitaxel cytotoxicity, and paclitaxel-induced apoptosis. Compared to drug sensitive MCF-7 cells, MCF-7ADR cells accumulated >6-fold less paclitaxel, were approximately 100-fold more resistant to killing by the drug, and were highly resistant to paclitaxel-induced apoptosis. In contrast, MCF-7ADR cells pretreated with valspodar were indistinguishable from drug-sensitive cells in their ability to accumulate paclitaxel, in their chemosensitivity to the drug, and in their ability to undergo paclitaxel-induced apoptosis. Valspodar, by itself, did not affect these parameters. This suggests that the enhancement of paclitaxel toxicity in MCF-7ADR cells involves a restoration of apoptosis and not solely through enhanced drug-induced necrosis. Morever, it appears that changes in the levels/activity of p53, the Fas receptor, Fas ligand, caspase-2, caspase 6, or caspase-8 activity have little effect on paclitaxel-induced cytotoxicity and apoptosis in human breast cancer cells. PMID- 10832592 TI - Weekly schedule of vinorelbine in pretreated breast cancer patients. AB - PURPOSE: In this phase II study, we explored tolerability and activity of vinorelbine administered according to a dose-dense weekly schedule with hematopoietic growth factor support in pretreated, advanced breast cancer patients. PATIENTS AND METHODS: From January 1994 to March 1996, 40 patients with metastatic breast cancer, pretreated with at least one prior anthracycline containing regimen, were entered into the study. PATIENT CHARACTERISTICS: median age 53 years (range 32-70); ECOG performance status 0-1: 34 patients, 2: 6 patients; dominant visceral metastatic disease: 15 patients, dominant non visceral: 25; anthracycline-refractory/resistant: 2 patients, sensitive: 38 patients. Six patients were treated as first-line therapy for metastatic disease and 34 in second- or subsequent lines. All patients received vinorelbine at the dose of 25 mg/m2/week as a short intravenous infusion, together with routine antiemetic medication. Granulocyte-colony stimulating factor (Lenograstim) at the dose of 150 microg/m2 subcutaneously on day 3 was included in the treatment schedule. RESULTS: The median number of treatment weeks was 23 (range: 4-24), with a delivered dose-intensity (DDI) of 23.8 mg/m2/week (range: 18.7-25, 95.2% of projected dose-intensity). Toxicity was mild, with non-complicated neutropenia being the main toxicity observed (grade 3-4 in 25% of the patients but only 2% of treatment weeks). Overall response rate was 52.5%, with complete responses in 12.5% of patients. Median duration of the response and median time to progression were 10 and 9 months, respectively. Median overall survival was 19 months. CONCLUSION: Dose-dense weekly vinorelbine is safe and effective with minimal toxicity in pretreated advanced breast cancer patients. PMID- 10832594 TI - Inhibition of HGF/SF-induced breast cancer cell motility and invasion by the HGF/SF variant, NK4. AB - NK4 is a variant form of HGF/SF, comprising the N-terminal and subsequent four kringle domains of mature HGF/SF. HGF/SF is a multifunctional cytokine that enhances the metastatic behaviour of tumour cells in vitro by stimulation of the c-met receptor tyrosine kinase and has been implicated in the development of tumour metastasis in vivo. The aims of this study were to further investigate the potential antagonistic effects of the recently described variant form of HGF/SF, NK4, on HGF/SF activity in breast cancer cells. All cell lines used expressed both the HGF/SF receptor gene and protein as shown by RT-PCR and Western blotting. NK4 inhibited HGF/SF-induced tumour cell invasion through an artificial basement membrane. Tumour cell motility and scattering induced by HGF/SF were also dramatically reduced by the inclusion of NK4. Immunoprecipitation studies revealed that NK4 inhibited the phosphorylation of the c-met receptor in response to HGF/SF. Treatment of these cells with NK4 alone did not have any significant effects on their metastatic behaviour. From this data we conclude that NK4 demonstrates significant antagonistic properties towards HGF/SF, inhibiting HGF/SF-stimulated breast tumour cell invasion, motility and migration. NK4 may therefore be of potential benefit in the development of anti-metastasis therapies. PMID- 10832595 TI - Administration of a targeted cytotoxic analog of luteinizing hormone-releasing hormone inhibits growth of estrogen-independent MDA-MB-231 human breast cancers in nude mice. AB - Receptor targeted chemotherapy is less toxic and more effective than conventional chemotherapy. Receptors for luteinizing hormone-releasing hormone (LH-RH) are found in about 50% of human breast cancers. Highly potent cytotoxic radical 2 pyrrolinodoxorubicin (AN-201) was linked to the agonistic analog [D-Lys6]LH-RH to form cytotoxic LH-RH analog AN-207. We evaluated whether AN-207 could be targeted to the hormone-independent MDA-MB-231 human breast cancers. Nude mice bearing MDA MB-231 tumors were injected i.v. with 250 nmol/kg doses of cytotoxic radical AN 201, cytotoxic LH-RH analog AN-207, the unconjugated mixture of AN-201 and carrier [D-Lys6]LH-RH, [D-Lys6]LH-RH alone and vehicle (control). The growth of MDA-MB-231 tumors in animals given a single dose of AN-207 was inhibited significantly (p = 0.01) for 3 weeks after injection, whereas tumors in all the other groups grew steadily. All cytotoxic compounds produced leukopenia, but the strongest lymphocyte suppression was caused by cytotoxic radical AN-201. Three weeks after treatment, the presence of mRNA for LH-RH receptors was demonstrated by RT-PCR in all the groups and radioreceptor assays demonstrated high-affinity binding sites for LH-RH on tumor cell membranes of control animals and those treated with AN-201, the carrier peptide alone or in combination with AN-201. At this time point binding assays did not reveal the expression of membrane proteins in tumors treated with AN-207, but 60 days after administration of AN-207, high affinity LH-RH binding sites were found again in MDA-MB-231 tumors. These results indicate that cytotoxic LH-RH analog AN-207 could be utilized for receptor targeted chemotherapy of breast cancers expressing receptors for LH-RH. PMID- 10832596 TI - Differential steroid hormone regulation of human glandular kallikrein (hK2) and prostate-specific antigen (PSA) in breast cancer cell lines. AB - We have investigated the steroid hormone regulation of human glandular kallikrein (hK2) and prostate-specific antigen (PSA) in the breast cancer cell lines BT-474, T-47D, MFM-223, MCF-7, ZR-75-1, MDA-MB-435, and BT-20. Using highly sensitive time-resolved fluorometric immunoassays, we were able to detect significant amounts of both kallikreins in tissue culture supernatants of BT-474, T-47D, and MFM-223 cells after hormonal stimulation. However, BT-474 cells produce much more hK2 than PSA, whereas the situation is reversed in T-47D cells. Furthermore, BT 474 cells produce, on absolute terms, about 500-1,000-fold more hK2 than T-47D cells. From all steroids tested, mibolerone, a synthetic non-metabolizable androgen, was the most potent stimulator for both kallikreins followed by the synthetic progestin norgestrel. Estradiol was able to induce production of small but significant amounts of hK2 and PSA in the BT-474 cell line, supporting the notion that there is a cross-talk between the estrogen and androgen hormone receptor signaling pathways. MFM-223 is an androgen responsive cell line, devoid of other steroid hormone receptors, which is also capable of producing hK2 and PSA but at much lower amounts. MCF-7 and ZR-75-1 cell lines failed to produce any protein, even though they have similar steroid receptor content as the BT-474 and T-47D cell lines. This was also the case for MDA-MB-435, a cell line rich in androgen receptors. Our data suggest that the expression of the hK2 gene in breast cancer cell lines is mainly under the control of androgens and progestins, similarly to PSA. These cell lines may represent good models for studying the differential expression of these two genes and for identifying cellular factors (e.g. co-activators/co-repressors), which may modify the potency of expression after hormonal stimulation. PMID- 10832597 TI - Metastatic breast cancer: clinical course, prognosis and therapy related to the first site of metastasis. AB - Although metastasis is a frequent event in breast cancer patients, insight into the clinical course, prognosis and therapy with respect to the site of the first metastases has been poor and contradictory in former investigations. Follow-up data from 648 patients with metastatic breast cancer were statistically analyzed. Patients with bone metastases at first relapse had better overall survival (median 71 vs. 48 months; p < 0.001) and survival after first metastases (median 24 vs 12 months; p < 0.001) than patients with visceral metastases at first relapse. Bone was the site of first metastasis in 46%, and 71% of patients with metastatic breast cancer developed bone metastases. The localization of the second metastatic site was of prognostic relevance in patients with first visceral metastases, but not in patients with first bone metastases. The presence of osseous metastases correlated significantly with estrogen and progesterone receptor positivity, tumor grading I/II and S-phase fraction <5%. The better prognosis of patients with bone metastases is not determined exclusively by hormone receptor status. The disease is significantly more stable in patients with first bone metastases than in those with first visceral metastases. PMID- 10832598 TI - Association between immunity and prognostic factors in early stage breast cancer patients before adjuvant treatment. AB - OBJECTIVE: The association of known prognostic factors with immune cell counts and beta2-microglobulin and soluble IL-2 receptor (sIL-2r) serum levels as markers of activation of the immune system was investigated in breast cancer. METHODS: Two hundred thirty five operated stage I and II breast cancer patients to receive adjuvant treatment in IBSCG trials were assessed in a cross-sectional study immediately before the first treatment. Leukocytes, lymphocytes and lymphocyte subset counts, beta2-microglobulin and sIL-2r serum levels were assessed as immunological parameters. Prognostic factors were tumor load, receptor status, patient characteristics, and contextual factors of the immune assessment (such as time of the day, time since surgery, type of surgery, concomitant medication, co-morbidity). RESULTS: In an operated early stage breast cancer patient population, tumor load was not associated with immune cell counts, beta2-microglobulin, or sIL-2r before adjuvant treatment. There was a pattern of association of prognostically favorable factors such as estrogen receptor (ER) positive tumor and older age with higher NK cell counts or with beta2 microglobulin or sIL-2r. In addition, immune cell counts and the markers of activation of the immune system were affected by several contextual factors, such as diurnal variability, time since surgery, type of surgery, and the intake of concomitant medication. CONCLUSIONS: The association of NK cell counts and beta2 microglobulin or sIL-2r serum levels with prognostically favorable factors such as ER positive tumor and older age supports the assumption that the immune system plays a role in the course of early breast cancer. The exact nature of this role requires further study. PMID- 10832599 TI - Kinetic effects of FPL 64176 on L-type Ca2+ channels in cardiac myocytes. AB - To characterize the effects of the Ca2+ channel agonist FPL 64176 on L-type Ca2+ current in isolated rat ventricular myocytes, certain of its effects were compared with those of a better known agonist, S (-) Bay K 8644. Both drugs enhance currents elicited by depolarizing pulses and enhance and slow the decay of tail currents elicited by subsequent repolarization. Both drugs shift the voltage dependence of activation and of inactivation approximately 10 mV in the negative direction, but FPL 64176 slows the rate of both activation and the decline of Ca2+ current during a depolarization, whereas Bay K 8644 accelerates the rate of current decay under the same conditions. In single channel studies in on-cell recording mode, FPL 64176 produced a great lengthening of the channel open time, produced very long openings when the channels were repolarized after a depolarizing stimulus, and had only modest effects on mean closed times and on first latency distributions. FPL 64176 and Bay K 8644 also had minimal effects on L-type channel "on" gating currents, while the "off" gating currents were slowed, particularly at positive potentials. However, the effects on gating currents were too small to account for the prolonged tails observed in FPL 64176. Once the channel is open, FPL 64176 slows transitions to closed or inactivated channel states. PMID- 10832600 TI - Involvement of soluble guanylate cyclase and calcium-activated potassium channels in the long-lasting hyporesponsiveness to phenylephrine induced by nitric oxide in rat aorta. AB - Excessive nitric oxide (NO) production by inducible NO synthase has been implicated in the hyporesponsiveness to vasoconstrictors present in septic shock. Here we show that a brief incubation (30 min) of rat aorta rings with NO donors renders the vessels hyporesponsive to phenylephrine for several hours. Contraction of rings without endothelium by phenylephrine (0.1 nM to 100 microM) was decreased by 50-60% after incubation (30 min) with sodium nitroprusside (3 300 microM) or S-nitroso-acetyl-D,L-penicillamine (SNAP; 70-200 microM). This decrease was characterized by reductions in maximal response and rightwards shifts of phenylephrine concentration/response curves, present even 130 min after NO donor removal. Soluble guanylate cyclase inhibitors methylene blue ( 10 microM) and 1H-(1,2,4)-oxadiazol-(4,3-a)quinoxalin-1-one (ODQ, 1 microM) or the potassium channel blockers TEA (tetraethylammonium; 10 mM) and charybdotoxin (100 nM) inhibited the hyporesponsiveness to phenylephrine induced by the NO donors. In contrast, 4-aminopyridine (1 mM) and glibenclamide (10 microM) had no effect. Our results show that incubation with NO donors reproduces the hyporesponsiveness to phenylephrine and that NO alone accounts for most, if not all, the refractoriness to vasoconstrictors present in septic shock. In addition, soluble guanylate cyclase activation and opening of potassium channels, more specifically the calcium-activated subtype, play a predominant role in this NO-induced hyporesponsiveness to phenylephrine in the rat aorta. PMID- 10832601 TI - In vivo electrophysiological examination of 5-HT2 responses in 5-HT2C receptor mutant mice. AB - The present study used 5-HT2C receptor mutant mice and their wild-type littermates to characterize the 5-HT2 receptor using the 5-HT2 agonists (+/-)-2 dimethoxy-4-iodoamphetamine hydrochloride (DOI) and 1-(3-chlorophenyl)piperazine (mCPP) applied locally in the orbitofrontal cortex (OFC) and head of the caudate nucleus. Microiontophoretically-applied 5-HT, DOI and mCPP induced current dependent inhibition of neuronal firing activity in both brain regions. There was no difference between 5-HT2C receptor mutants and wild-type mice in the ability of 5-HT or DOI to inhibit neuronal firing at any current used. In contrast, there was a reduced ability of mCPP to inhibit firing activity in the OFC when ejected at 10 nA. Unexpectedly, there was a small but significant increase in mCPP induced inhibition in the caudate nucleus of mutant mice. In the OFC, the 5-HT2A antagonist MDL 100907 (2 mg/kg, i.p.) significantly antagonized the effect of both DOI and mCPP. In contrast, the non-selective 5-HT antagonist clozapine (10 mg/kg, i.p.) significantly antagonized only mCPP in the wild-type mice. However, neither MDL 100907 nor clozapine antagonized DOI or mCPP in the caudate nucleus. Finally, it required significantly less quisqualate to activate neurons in the 5 HT2C receptor mutants than in the wild-type mice, suggesting that 5-HT2C receptors serve a tonic inhibitory role in membrane excitability. The present results indicate that the inhibitory action of DOI is predominantly mediated by the 5-HT2A receptor in the OFC. mCPP, when applied locally, inhibits OFC firing activity by acting on both 5-HT2A and 5-HT2C receptors. However, DOI and mCPP might be acting in the caudate nucleus through an atypical 5-HT2 receptor yet to be characterized. PMID- 10832602 TI - Inhibitory effect of PACAP(6-38) on relaxations induced by PACAP, VIP and non adrenergic, non-cholinergic nerve stimulation in the guinea-pig taenia caeci. AB - The effect of the pituitary adenylate cyclase activating polypeptide (PACAP) receptor antagonist PACAP(6-38) on the relaxant response to exogenous PACAP, vasoactive intestinal polypeptide (VIP) and nonadrenergic, non-cholinergic (NANC) nerve stimulation was tested in the guinea-pig taenia caeci, in the presence of atropine (10(-6) M) and guanethidine (3x10(-6) M). PACAP(6-38) (3x10(-6) M) strongly inhibited sub-maximal relaxations evoked by exogenous PACAP (1-3x 10(-8) M) or VIP (10(-8) M), but not those due to isoprenaline (4-8x10(-8) M) or ATP (10(-6) M). PACAP(6-38) caused a small but significant (approximately 20%) inhibition of the NANC relaxation due to electrical field stimulation (1 Hz or 10 Hz for 20 s). At these frequencies PACAP(6-38) caused no inhibition of the NANC relaxation in the presence of the P2 purinoceptor antagonist pyridoxal-phosphate 6-azophenyl-2',4'-disulphonic acid (PPADS; 5x10(-5) M), or PPADS plus the NO synthase blocker NG-nitro-L-arginine (L-NOARG; 10(-4) M); in preparations pretreated with L-NOARG (10(-4) M) alone PACAP(6-38) retained its inhibitory effect. The PPADS- and L-NOARG-resistant NANC relaxation with 10 Hz electrical stimulation was blocked by apamin (10(-7) M); it was not significantly modified by the tachykinin receptor antagonist spantide (10(-5) M). Tachyphylaxis to PACAP(1-27) (10(-7) M for 10 min) strongly inhibited the relaxation due to PACAP(1-38) (1-3x10(-8) M) and reduced electrical stimulation-evoked relaxations by half. The putative VIP antagonist VIP(10-28) (10(-5) M) failed to significantly reduce the relaxant action of exogenous VIP (1-3x10(-8) M). Relaxation induced by PACAP(1-38) (1-2x10(-8) M) was not influenced by a mixture of PPADS (5x10(-5) M) and L-NOARG (10(-4) M). It is concluded that: (a) PACAP(6 38) is a VIP/PACAP antagonist in the guinea-pig taenia caeci; (b) a release of a VIP/PACAP-like substance from enteric nerves is involved in the NANC relaxation in this preparation, but its contribution is relatively small and seems to depend on the functional integrity of the PPADS-sensitive inhibitory mechanism; (c) the PPADS- plus L-NOARG-resistant NANC relaxation probably involves apamin-sensitive K+ channels. PMID- 10832603 TI - Characterization of dopaminergic compounds at hD2short, hD4.2 and hD4.7 receptors in agonist-stimulated [35S]GTPgammaS binding assays. AB - Dopamine receptor agonists and antagonists have been extensively characterized in radioligand binding assays; only a limited number of laboratories have characterized them using a functional assay at multiple receptor subtypes. Experiments were designed to assess four agonists and seven antagonists at three cloned human dopamine receptors using agonist-stimulated [35S]GTPgammaS binding assays in membranes to quantify the initial cellular event following ligand/receptor interaction. In this model there is constitutive G protein activity (agonist-independent [35S]GTPgammaS binding) and potentially constitutive dopamine receptor activity. Thus, discrimination between silent antagonists, partial agonists and inverse agonists is theoretically possible. It was anticipated that distinctions could be made regarding efficacy of the seven receptor antagonists to provide insight regarding the therapeutic use of antipsychotic drugs. In membranes prepared from CHO cells transfected to express high densities of human D2short, D4.2 or D4.7 receptors, the dopamine receptor agonists apomorphine, pergolide, quinelorane and quinpirole produced concentration-dependent increases in agonist-stimulated [35S]GTPgammaS binding. At the hD2short receptor, pergolide and apomorphine were essentially equipotent and more potent than quinelorane and quinpirole; all four agonists displayed similar efficacy at this receptor. At the hD4.2 and the hD4.7 receptors apomorphine was the most potent and pergolide the least efficacious of the four drugs. The ability (both potency and efficacy) of clozapine, haloperidol, olanzapine, quetiapine, risperidone, spiperone and ziprasidone to block apomorphine-stimulated [35S]GTPgammaS binding and alter basal [35S]GTPgammaS binding was also assessed. All of the antagonists inhibited apomorphine stimulated [35S]GTPgammaS binding with potencies (Kb values) similar to and in rank order consistent with their affinities reported in the literature using radioligand binding assays. Additionally, none of the antagonists altered basal, agonist-independent [35S]GTPgammaS binding, thus they behaved as pure, silent antagonists at D2short, D4.2 and D4.7 receptors under our conditions. In summary, the data suggest that therapeutic distinctions between typical and atypical antipsychotic drugs cannot be made based on their function at D2short, D4.2 and D4.7 subtypes of dopamine receptors. PMID- 10832604 TI - Characterization of the transport of the organic cation [3H]MPP+ in human intestinal epithelial (Caco-2) cells. AB - The aim of this study was to characterize the transport of organic cations at the intestinal level, by studying the characteristics of the transport of 1-methyl-4 phenylpyridinium (MPP+) in Caco-2 cells. Transepithelial flux as well as cellular accumulation of [3H]MPP+ were quantitatively similar when substrate was applied from the basolateral or apical cell membrane. Verapamil (100 microM) and rhodamine123 (10 microM) significantly reduced [3H]MPP+ transepithelial flux in the apical-to-basolateral direction. When cells were grown on plastic supports, [3H]MPP+ was rapidly accumulated in the cells, both by saturable and nonsaturable mechanisms. The kinetic parameters of the saturable component were: Km: 449 microM and Vmax: 2,249 pmol per mg protein and 5 min. Uptake of [3H]MPP+ was metabolic energy-dependent and Na+-, pH- and potential-independent. It was inhibited by several organic cations (verapamil, rhodamine123, daunomycin, vinblastine, tetrabutylammonium and vecuronium) but not by others (tetraethylammonium and N-methylnicotinamide). Decynium22 and corticosterone inhibited [3H]MPP+ uptake into the cells. The P-glycoprotein antibody UIC2 (20 microg/ml) had no effect. In conclusion, [3H]MPP+ is efficiently transported by Caco-2 cells in both basolateral-to-apical (secretion) and apical-to-basolateral (absorption) directions. Absorption of [3H]MPP+ at the apical membrane seems to occur through a carrier-mediated mechanism belonging to the Amphiphilic Solute Facilitator (ASF) family of transporters, but distinct from the known members of this family. PMID- 10832605 TI - The influence of sex hormones on vascular responses in the aorta of streptozotocin-diabetic male rats. AB - Diabetes mellitus reduces gender-related differences in the prevalence of cardiovascular disease by fading the vascular protective effects afforded by estrogen in females. However, the impact of estrogen treatment on and the contribution of androgens to vascular function in vessels from male diabetics are largely unknown. We investigated the effects of androgen deficiency and in vivo estrogen treatment by assessing the responsiveness to a number of vasoactive agents and the formation of eicosanoid mediators in aortic rings from intact and castrated streptozotocin-diabetic rats which had been implanted with 17beta estradiol (E2) or its vehicle for 5 days. Castration was found to attenuate contractility to noradrenaline, to enhance tone-related release of NO, as shown by curves for N-methyl-L-arginine and superoxide dismutase (SOD), and to increase endothelium-dependent relaxation to carbachol and histamine, compared with intact animals. Smooth muscle sensitivity to exogenous NO and platelet thromboxane A2 production were unchanged but prostacyclin release by aortic tissue dropped by about 40% following castration. Treatment with E2 to intact animals still attenuated contractility to noradrenaline and potentiated relaxation to SOD and histamine but affected no other parameters. In contrast, when E2 was administered to castrated animals, responses to SOD, carbachol and histamine were significantly impaired. Thus, androgen deprivation appears to improve vascular function in male diabetic rats, whereas E2 treatment exerts some beneficial effects in intact, but not in castrated animals. Our findings therefore provide new insights into the role of sex hormones in the development of diabetic vascular complications. PMID- 10832606 TI - Serotonergic mechanisms involved in the attentional and vigilance task performance of rats and the palliative action of aniracetam. AB - Central serotonergic systems play an important role in regulating mood/emotion, cognition, sleep and wakefulness, appetite and locomotion and body temperature via multiple receptor subtypes. Among them, 5-HT1A and 5-HT2A/2C receptors have opposite effects with respect to certain functions. The aim of the present study was to compare the effects of 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT), a selective 5-HT1A receptor agonist, and 1-(2,5-dimethoxy-4-iodophenyl)-2 aminopropane (DOI), a selective 5-HT2A/2C receptor agonist, on the performance of middle-aged rats in a two-lever choice reaction task that assessed attention and vigilance functions. We also examined the effects of aniracetam, a cognition enhancer, and its major metabolites on the induced performance impairments. 8-OH DPAT (0.3 mg/kg s.c.) reduced response speed and choice accuracy and increased response omission with a reduction of task-associated motor activity without inducing motor inability or motivational changes. These findings indicate a specific disturbance of attentional and vigilance processes. DOI caused similar impairments at the highest dose tested (3 mg/kg s.c.); at a lower dose (1 mg/kg s.c.), however, it selectively attenuated the response speed, suggesting a selective attention deficit. (-)-Alprenolol, a non-selective 5-HT1A receptor antagonist, and ritanserin, a preferential 5-HT2A receptor antagonist, blocked the 8-OH-DPAT- and DOI-induced performance impairments respectively. Aniracetam ameliorated all the performance deficits, and the metabolites N-anisoyl-GABA and 2-pyrrolidinone partially mimicked the aniracetam effect in the 8-OHDPAT-induced attentional and vigilance impairments. Nefiracetam, another cognition enhancer, improved only the 8-OH-DPAT-induced impairments. Each compound tested alone had no effect on task performance. These results indicate that both serotonergic regulations, possibly via presynaptic 5-HT1A receptors and more likely via postsynaptic 5-HT2A receptors, lead similarly to attention deficits. PMID- 10832607 TI - An increase of the native interstitial adenosine concentration during histidine application. AB - We examined whether histidine can increase the production of interstitial adenosine via noradrenaline (NA) release-mediated activation of ecto-5' nucleotidase in the ventricular myocardium, with use of microdialysis techniques in in situ rat hearts. The microdialysis probe was implanted in the left ventricular myocardium of anesthetized rat hearts and the tissue was perfused with Tyrode's solution containing adenosine 5'-monophosphate (AMP) through the dialysis probe at a rate of 1.0 microl/min. Adenosine in the dialysate collected during perfusion with Tyrode's solution containing 100 microM AMP (through the probe) originated from the hydrolysis of AMP catalyzed by endogenous ecto-5' nucleotidase, so that the level of adenosine reflected the activity of ecto-5' nucleotidase in this tissue. In the presence of NA (10 microM), histidine, a scavenger of highly active singlet oxygen (1O2), significantly increased concentration of adenosine. Histidine (5-50 mM) increased the level of AMP-primed dialysate adenosine in a concentration-dependent manner. When histidine (25 mM) was infused to rat myocardium, small increase in the levels of adenosine were observed. However, when corresponding experiments were performed with NA (10 microM)-pretreated animals, a marked elevation of the level of adenosine in rat heart dialysate was obtained. To confirm the possible mechanism of interaction between 1O2 and NA, we examined the effect of histidine in ischemic-reperfused rat hearts. In the presence of histidine (25 mM), a marked elevation of NA and adenosine was observed. However, when corresponding experiments were performed with reserpinized rat hearts, the elevation of both NA and adenosine was not observed in ischemia-reperfused rat hearts. These results indicate that histidine increases interstitial adenosine concentration via NA release-mediated activation of ecto-5'-nucleotidase. PMID- 10832608 TI - Differential actions of anandamide, potassium ions and endothelium-derived hyperpolarizing factor in guinea-pig basilar artery. AB - Vasodilator responses to anandamide (arachidonylethanolamide) and potassium ions were compared with those mediated by endothelium-derived hyperpolarizing factor (EDHF) in guinea-pig isolated basilar artery contracted with prostaglandin F2alpha. In this artery, EDHF-mediated responses can be evoked by acetylcholine in the presence of both indomethacin (10 microM) and NG-nitro-L-arginine (0.3 mM). In endothelium-denuded arterial segments, which failed to respond to acetylcholine, anandamide was still able to evoke a complete relaxation. Anandamide (10 microM) did not affect the resting membrane potential, whereas acetylcholine (10 microM) hyperpolarized the smooth muscle cells by 23 mV in the presence of indomethacin and NG-nitro-L-arginine. Pre-treatment with capsaicin (10 microM) or resiniferatoxin (0.1 microM) abolished the anandamide-induced relaxation, but had no effect on the EDHF-mediated relaxation induced by acetylcholine. Treatment with a mixture of the calcium-sensitive potassium channel inhibitors, apamin and charybdotoxin, which abolishes EDHF-mediated relaxation in this artery, did not affect the relaxation evoked by anandamide. The additional presence of glibenclamide or ciclazindol, inhibitors of ATP sensitive and voltage-dependent potassium channels, also had no effect on the anandamide-induced relaxation. Increasing the potassium ion concentration by 2-10 mM induced inconsistent vasodilator responses. However, re-admission of potassium ions to preparations incubated in potassium-free solution elicited almost complete and sustained relaxations. A short incubation period with ouabain (10 microM for 10 min) or cooling (18-22 degrees C) abolished these responses, whereas the acetylcholine-induced relaxation in the presence of indomethacin and NG-nitro-L-arginine was unaffected (ouabain) or partially reduced (cooling). The anandamide-induced relaxation was also abolished by ouabain and cooling. Furthermore, ouabain inhibited the vasodilator response to capsaicin, but not that to calcitonin gene-related peptide (CGRP), and per se evoked a release of CGRP from the artery. The gap junction uncoupler, 18alpha-glycyrrhetinic acid (100 microM), affected neither the EDHF-mediated relaxation induced by acetylcholine nor the vasodilator responses to anandamide and potassium ions. Thus, EDHF-mediated vasorelaxation in the guinea-pig basilar artery does not seem to involve Na+/K+-ATPase, sensory nerves or gap junctions. These results indicate that EDHF is neither anandamide nor potassium ions in this artery. PMID- 10832609 TI - Role of supraspinal tachykinins for micturition in conscious rats with and without bladder outlet obstruction. AB - In order to clarify the role of supraspinal tachykinins in volume-induced micturition and in bladder hyperactivity secondary to bladder outlet obstruction, conscious, normal, female Sprague-Dawley rats were investigated cystometrically before and after intracerebroventricular administration of RP 67,580, a selective antagonist of neurokinin (NK)-1 receptors and/or SR 48,968, a selective antagonist of NK-2 receptors. In normal rats, RP 67,580 or SR 48,968, at a dose of 2 nmol, caused no marked changes in cystometric parameters. Higher doses (up to 20 nmol) caused dose-dependent decreases in micturition pressure and increased bladder capacity, micturition volume and residual urine. A combination of the two drugs, each at a dose of 2 nmol, significantly decreased micturition pressure and increased bladder capacity. In rats with bladder outlet obstruction, the antagonists suppressed micturition dose-dependently, producing urinary retention in two out of eight rats already at a dose of 2 nmol. At a dose of 20 nmol, dribbling incontinence, due to urinary retention, was seen in five out of ten rats. A combination of the two drugs (2 nmol of each drug) caused urinary retention in three out of nine animals and significantly increased bladder capacity, micturition volume and residual volume. The results suggest that outflow obstruction in rats increases the effects of tachykinins in supraspinal structures involved in micturition, and that antagonism of supraspinal NK receptors may depress the micturition reflex. Whether or not this implies that supraspinal NK-receptors can be targets for drugs aimed for inhibiting bladder hyperactivity in humans should be explored. PMID- 10832610 TI - Antagonist properties of the novel antipsychotic, S16924, at cloned, human serotonin 5-HT2C receptors: a parallel phosphatidylinositol and calcium accumulation comparison with clozapine and haloperidol. AB - The novel benzopyranopyrrolidine and potential antipsychotic, S16924 ((+)-2-[[1 [2-(2,3-dihydrobenzo[ 1,4] dioxin-5-yloxy)-ethyl]-pyrrolidin-3yl]]-1-(4-fluoro-ph enyl)-ethanone), displays marked affinity for serotonin (5-HT)1A, 5-HT2A and dopamine D2 receptors. Herein, we show that it also possesses high affinity for the cloned, INI isoform of h5-HT2c receptors (pKi=8.28) stably expressed in CHO cells. Similarly, clozapine (8.04) was a potent ligand, whereas haloperidol (<6.0) showed low affinity. As demonstrated by fura2-detection, S16924 concentration-dependently abolished (pKb=7.93) the 5HT-induced elevation in intracellular levels of Ca2+ ([Ca2+]i) in a CHO cell line stably expressing the INI isoform of 5-HT2c receptors. Further, as determined by depletion of membrane bound levels of pre-labelled [3H]phosphatidylinositols ([3H]PI), S16924 concentration-dependently, surmountably and competitively blocked the activation of phospholipase C by 5-HT. This action was expressed with a pA2 of 7.89 according to Schild analysis. Clozapine likewise inhibited 5-HT-induced alterations in [Ca2+]i and [3H]PI levels with pKbs of 7.43 and 7.84, respectively, whereas haloperidol was inactive (<5.0 in each case). Applied alone, S 16924, clozapine and haloperidol modified levels of neither [Ca2+]i nor [3H]PI. In conclusion, in analogy to clozapine, and in contrast to haloperidol, S16924 behaves as a potent and competitive antagonist at h5-HT2c receptors, the blockade of which may contribute to its distinctive functional profile of activity. PMID- 10832611 TI - Cloning, expression, functional coupling and pharmacological characterization of the rat dopamine D4 receptor. AB - It has been difficult to observe functional coupling of the D4 receptor to second messenger systems and a robust functional assay system for this receptor is still lacking. In the present study, the rat dopamine D4 receptor was cloned from rat retina. Sequence comparison revealed identity with the published sequence of Ashgari and co-workers, including the two amino acid insertions (V-Q) at position 92 which are not present in the published sequence of O'Malley and coworkers. The rat dopamine D4 receptor was stably expressed in Chinese hamster lung fibroblast CCL39 cells. [3H]spiperone saturation binding yielded a Bmax of 2,370+/-546 fmol/mg protein and a pKD of 8.74+/-0.14 (n=4). Forskolin-stimulated cAMP accumulation was inhibited by dopamine (Emax 61+/-1% inhibition of forskolin stimulated levels, pEC50 7.33+/-0.06, n=23). A similar concentration-dependent inhibition was observed with the dopamine D2-like receptor agonists quinpirole and 7-OH-DPAT which elicited nearly the same Emax as dopamine. By contrast, apomorphine and a number of compounds with reported affinity for human dopamine D4 receptors (PD168077, U-101958, SDZ GLC 756, L-745,870 and NGD 94-1) behaved as partial agonists (Emax ranging between 26% and 56% of that of dopamine). The agonist effect of dopamine was completely blocked by preincubation with pertussis toxin, no further accumulation of cAMP above the forskolin-stimulated levels being observed. Antagonist pKB-values obtained against dopamine in this system were: 8.55+/-0.19 (n=3) for the partial agonist L-745,870, 8.38+/-0.23 (n=5) for spiperone, 7.18+/-0.17 (n=4) for haloperidol, 7.04+/-0.13 (n=4) for clozapine and <6 for raclopride. Other functional assays applicable were stimulation of [35S]GTPgammaS binding, extracellular acidification rate and a serum-responsive element using luciferase expression as a reporter gene. However, the receptor did not couple to phosphatidylinositol turnover or to intracellular Ca2+. Thus, expression of the rat dopamine D4 receptor in CCL39 cells provided several functional assay systems, of which inhibition of cAMP appeared to be the most robust one. These functional models can be used to evaluate the activity of compounds at the rat dopamine D4 receptor. PMID- 10832612 TI - In vitro characterization of J-113397, a non-peptide nociceptin/orphanin FQ receptor antagonist. AB - The lack of availability of a selective, highly potent, competitive antagonist for the nociceptin receptor (OP4) devoid of residual agonistic activity has hampered studies in this area. We report here the in vitro pharmacological properties of the novel non-peptide OP4 antagonist, J-113397, which was recently discovered by Banyu Pharmaceutical investigators. The compound was synthesized as a racemic mixture in our laboratories. J-113397 was shown to antagonize (pA2 7.52) the nociceptin-induced inhibition of cAMP formation in cells expressing the recombinant human OP4 receptor (CHOhOP4) and to displace [125I]Tyr14nociceptin from CHOhOP4 membranes with a pKi of 8.56. It also competitively antagonized the contractile actions of nociceptin in the mouse colon (pA2 8.07) and the inhibitory effect of nociceptin in electrically stimulated preparations such as the mouse vas deferens (pA2 7.85), the guinea pig ileum (7.75), and the rat vas deferens (7.77). At high concentrations (10 microM), the compound was devoid of agonist activity in the mouse vas deferens and CHOhOP4, while it contracted the mouse colon and increased the twitch response of the rat vas deferens, and produced a naloxone-sensitive inhibition of the electrically evoked twitches in the guinea pig ileum. pA2 values for the new antagonist against deltorphin I in the mouse vas deferens (OP1 receptors), or against dermorphin in the guinea pig ileum (OP3 receptors), etorphine in the rat vas deferens (OP receptors), U69593 in the rabbit vas deferens (OP2 receptors) and endomorphin 1 in the mouse colon (OP3 receptors) were lower than 6. Taken together, these data indicate that J 113397 is a high-affinity, selective and competitive antagonist of the OP4 receptor; this novel pharmacological tool will be of great value in studies directed at evaluating the physiological roles of the nociceptin/OP4 system. PMID- 10832613 TI - [3H]S33084: a novel, selective and potent radioligand at cloned, human dopamine D3 receptors. AB - The novel, selective dopamine D3 receptor antagonist, S33084 [(3aR,9bS)-N[4-(8 cyano- 1,3a,4,9b-tetrahydro-3H-benzopyrano[3,4-c]pyrrole-2-yl)-butyl] (4 phenyl)benzamide], was tritium-labelled to 59 Ci/mmol specific activity. Determination of association and dissociation rate constants at recombinant, human (h) D3 receptors stably expressed in Chinese hamster ovary (CHO) cells yielded a Kd value (0.16 nM) comparable to that observed in saturation binding experiments (0.17 nM). The competition binding profile of [3H]S33084 with diverse D3 receptor agonists and antagonists correlated highly (0.99) with that of [3H]spiperone. In conclusion, [3H]S33084 is a highly potent and selective radioligand at dopamine D3 receptors, which should be of considerable use for their characterisation. PMID- 10832614 TI - Serum immunoglobulins and lymphocyte subset distributions in children and adults living in communities assessed for lead and cadmium exposure. AB - This study assessed the impact of environmental cadmium and lead exposure on the immune system of more than 2000 children and adults. Serum immunoglobulins [immunoglobulins (Ig) A, G, and M] and peripheral blood lymphocyte phenotypes (T cells, B cells, NK cells, and CD4/CD8 subsets) were measured in a total of 2041 children and adults who lived either in sites with elevated soil levels of cadmium and lead (n = 1561) or in comparison communities (n = 480). The blood lead and urine cadmium levels of participants were somewhat higher than national averages. Mean blood lead levels were 7 microg/dl for participants aged 6-35 mo; 6 microg/dl for participants aged 36-71 mo, 4 microg/dl for participants aged 6 15 yr; and 4.3 microg/dl for participants aged 16-75 yr. Multivariate analysis indicated no marked differences in any of the immune marker distributions attributed to lead for adults or children over 3 yr of age. However, in children under age 3, increased blood lead levels, principally those over 15 microg/dl, were associated with increases in IgA, IgG, IgM, and circulating B lymphocytes. Among adults, urine cadmium levels over 1.5 microg/g were associated with higher levels of IgA and circulating B lymphocytes. No evidence of immunosuppression was noted. The findings of potential immunologic effects at lead levels > 15 microg/dl in young children and at urine cadmium levels > 1.5 microg/g in adults are interesting, but too few participants had these high levels to delineate a threshold. Therefore, we find these results intriguing, but requiring confirmation in populations with higher exposure levels. PMID- 10832615 TI - Calcium and magnesium in drinking water and risk of death from prostate cancer. AB - The possible association between the increased risk of prostate cancer and the levels of calcium and magnesium in drinking water from municipal supplies was investigated in a matched cancer case-control study in Taiwan. All eligible prostate-cancer deaths (682 cases) of Taiwan residents from 1987 through 1993 were compared with deaths from other causes (682 controls). The levels of calcium and magnesium in the drinking water of these residents were also determined. Data on calcium and magnesium levels in drinking water throughout Taiwan were obtained from the Taiwan Water Supply Corporation (TWSC). The control group consisted of people who died from other causes, and the controls were pair-matched to the cases by year of birth and death. The adjusted odds ratios for prostate cancer death for those with higher magnesium levels in their drinking water, as compared to the lowest tertile, were 0.73 (95% CI = 0.51-1.03) and 0.64 (95% CI = 0.43 0.96), respectively. The adjusted odd ratios for the relationship between calcium levels in drinking water and prostate cancer were not statistically significant. The results of the present study show that there may be a significant protective effect of magnesium intake from drinking water and other dietary sources against the risk of prostate cancer development. PMID- 10832616 TI - Silica-induced nuclear factor-kappaB activation: involvement of reactive oxygen species and protein tyrosine kinase activation. AB - Nuclear factor-kappaB (NF-kappaB) is a multiprotein complex that may regulate a variety of inflammatory cytokines involved in the initiation and progression of silicosis. The present study documents the ability of in vitro silica exposure to induce DNA-binding activity of NF-kappaB in a mouse peritoneal macrophage cell line (RAW264.7 cells) and investigates the role of reactive oxygen species (ROS) and/or protein tyrosine kinase in this activation. In vitro exposure of mouse macrophages to silica (100 microg/ml) resulted in a twofold increase in ROS production, measured as the generation of chemiluminescence (CL), and caused activation of NF-kappaB. Silica-induced CL was inhibited 100% by superoxide dismutase (SOD) and 75% by catalase, while NF-kappaB activation was inhibited by a variety of antioxidants (catalase, superoxide dismutase, alpha-tocopherol, pyrrolidine dithiocarbamate, or N-acetylcysteine). Further evidence for the involvement of ROS in NF-kappaB activation is that 1 mM H2O2 enhanced NF kappaB/DNA binding and that this activation was inhibited by catalase. Specific inhibitors of protein tyrosine kinase, such as herbimycin A, genistein, and AG 494, prevented NF-kappaB activation in silica-treated cells. Genistein and AG-494 also reduced NF-kappaB activation in H2O2-treated cells. Results confirm that tyrosine phosphorylation of several cellular proteins (approximate molecular mass of 39, 58-70, and 103 kD) was increased in silica-exposed macrophages and that genistein inhibited this silica-induced phosphorylation. In contrast, inhibitors of protein kinase A or C, such as H89, staurosporin, calphostin C, and H7, had no marked inhibitory effect on silica-induced NF-kappaB activation. The results suggest that ROS may play a role in silica-induced NF-kappaB activation in macrophages and that phosphorylation events mediated by tyrosine kinase may be involved in this activation. PMID- 10832617 TI - Mineral and/or metal content as critical determinants of particle-induced release of IL-6 and IL-8 from A549 cells. AB - Mineral particles in occupational exposure and ambient air particles may cause adverse health effects in humans. In this study the ability of different stone quarry particles to induce release of the proinflammatory cytokines interleukin-6 (IL-6) and interleukin-8 (IL-8) from human epithelial lung cells (A549) was investigated. Size distribution within the PM10 fractions was quite similar for all particle samples, whereas mineral content and metal composition differed. Particles, containing minerals such as quartz, amphibole, chlorite, and epidote, induced a marked increase in IL-6 and IL-8 release. Particles composed mainly of plagioclase were much less effective. The most potent particle samples exhibited a relatively high content of transition metals such as iron. Exposure to identical masses or surface areas resulted in the same order of potency among the different particle samples. Significant cytotoxicity was observed only at higher concentrations of particle exposure. Thus, mineral composition and/or metal contents of particles from different stone quarries were critical determinants for the marked differences in potency to induce cytokine responses in human epithelial lung cells. PMID- 10832619 TI - 3D assessment of myocardial perfusion parameter combined with 3D reconstructed coronary artery tree from digital coronary angiograms. AB - In patients with coronary artery disease coronary angiography plays an important role in the clinical decision-making process. However, it has been recognized that no simple relation exists between the visually or quantitatively evaluated severity of coronary artery stenoses and its effects on regional myocardial perfusion. This paper describes for the first time the development and application of a 3D technique that visualizes and quantifies regional myocardial perfusion parameters from biplane coronary angiograms by using the impulse response analysis technique. The 3D reconstructed coronary tree is automatically superimposed on the 3D perfusion image to generate and visualize an 'integrated' 3D image. The preliminary results in patients with critical coronary artery stenoses indicate that our combined 3D fusion image provides flow information from the major coronary arteries. This 3D fusion image may provide useful information in the management of patients with coronary artery disease. PMID- 10832618 TI - LLC-PK1 cells as a model for renal toxicity caused by arsine exposure. AB - The mechanisms of arsine (AsH3) toxicity are not completely understood. Studies were undertaken to determine AsH3 and arsenite [As(III)] toxicity in a renal tubular epithelial cell line to model kidney dysfunction caused by AsH3 exposure. The hypothesis was that As(III) is the toxic metabolite responsible for the renal toxicity of AsH3. There was a concentration- and time-dependent toxic response after As(III) incubation. As(III) produced significant LDH leakage as early as 1 h and intracellular potassium loss at 5 h. AsH3 produced no changes in these parameters. AsH3 affected neither potassium nor LDH levels over 24 h and up to 1 mM AsH3 concentration. In this system, As(III) induced LDH leakage before K+ loss. Oxidative stress-like toxicity effects were also studied by determining levels of glutathione (GSH), glutathione disulfide (GSSG), and heat-shock protein 32 (Hsp32) levels. GSH levels were not markedly affected by any arsenical over a 6-h period or up to 100 microM concentration of the arsenical. However, 100 microM AsH3 significantly increased GSSG levels as early as 30 min and reached a maximum at 2.5 h. Incubation with 10 microM AsH3 was sufficient to significantly increase GSSG levels. As(III) had no marked effect on GSSG. Both arsenicals (50 microM) produced a slight increase (about threefold) in Hsp32 levels after 4-h incubation. These results showed that unchanged AsH3 produced oxidative stress like toxic effects without producing cell death. However, similar As(III) concentrations induced the stress response and were toxic to the cells. These data indicated that AsH3 is not directly toxic to LLC-PK1 cells. PMID- 10832620 TI - Computerized left ventricular pressure-volume relationships (pV-loops) using disposable angiographic tip transducer pigtail catheters. AB - Left ventricular pressure-volume relationships expressed as pV loops could yield important hemodynamic information in the cardiac catheterization laboratory. Many clinical situations might benefit from a quantitative assessment of left ventricular function. Potential applications of pV loops include the assessment of vasoactive and inotropic drugs, balloon valvuloplasty, coronary angioplasty, and surgical treatment of valvular heart disease. For many years the clinical use of pV loops has been hindered by logistical difficulties. The ability to merge on line concurrent digital imaging data for computation of left ventricular volume and digital left ventricular pressure wave forms obtained from high fidelity tip transducer angiocatheters has allowed us to develop a technique which can generate pV loops during cardiac catheterization procedures. The method offers an automated measurement of left ventricular volume independent of edge detection or an interactive technique for tracing endocardial borders by a trained operator. Illustrative case studies are included to demonstrate the potential of the method during ventricular angiographic procedures. Implementation and computational time requirements of the method are discussed. The concept and the value of pV loop generation to study left ventricular performance has been known for many years. Combining digital imaging and digital physiologic data obtained with disposable tip-transducer angiocatheters with modern networking technology, the technique can more easily be applied to catheterization procedures and could enhance invasive hemodynamic assessment of left ventricular function. PMID- 10832621 TI - Mechanical properties and imaging characteristics of remanufactured intravascular ultrasound catheters. AB - Intravascular ultrasound (IVUS) as a routine device in interventional cardiology is handicapped by its high price. 19 factory-made, 'remanufactured' IVUS catheters which consist of sterilized, used phased-array IVUS transducers inserted into a new catheter shaft were compared with 23 new IVUS catheters. 3 mechanical and 4 imaging characteristics were assessed on a 5 point scale (1 = unacceptable, 5 = excellent). Mechanical as well as imaging properties of 'remanufactured' IVUS catheter were comparable to new catheters with excellent ratings for each of the evaluated characteristics in 38 to 94% of 'remanufactured' catheters and 50 to 96% of new catheters. The initial failure rate for 'remanufactured' IVUS catheters was 31.6% vs. 4.3% for new catheters (P < 0.05). Overall failure rate was 47.3% for "remanufactured" catheters vs. 8.7% for new catheters (P < 0.05). The failure was due to an electronic connecting problem occurring during mechanical stress to the IVUS catheter. In conclusion, 'remanufactured' IVUS catheters offer mechanical and imaging characteristics which are comparable to new catheters. Improvements in the 'remanufacturing' process to resolve the high rate of electronic connecting problems may make this a promising approach to substantially lower the price of IVUS catheters. PMID- 10832622 TI - Echocardiographic evaluation of left ventricular function in pure mitral stenosis. AB - Echocardiographic evaluation of left ventricular function was performed in 22 cases of pure rheumatic mitral stenosis and 22 age matched normal persons. Cases with any evidence of rheumatic activity in the preceding six months, those with gross tricuspid regurgitation and paradoxical movement of the interventricular septum and cases with atrial fibrillation were excluded. None of the patients showed systolic left ventricular dysfunction. Left ventricular end diastolic dimension was also not affected. Echocardiographic parameters did not have any relation for mitral valve area. Our observations show that mitral stenosis per se does not affect left ventricular function. PMID- 10832623 TI - Imaging of an aneurysm of the sinus of Valsalva with transesophageal echocardiography, contrast angiography and MRI. AB - A sinus of Valsalva aneurysm is an uncommon congenital defect, which requires appropriate diagnosis with either echocardiography, magnetic resonance imaging or contrast angiography. Treatment consists of aortic valve repair. We describe a young woman with an aneurysm of the non-coronary sinus of Valsalva, an atrial septal defect and pulmonary insufficiency. The different imaging techniques and possibilities of surgical correction are described. PMID- 10832624 TI - Reduced early diastolic inflow velocities in the antero-posterior transverse direction in the left ventricle of patients with dilated cardiomyopathy. AB - In patients with dilated cardiomyopathy (DCM), the left ventricular (LV) inflow jet is narrow and has a high pressure gradient. A pulsed Doppler restrictive transmitral flow pattern is a characteristic feature of severe left ventricular disease. However, Doppler flow analysis is limited by the angle between the blood flow jet and the ultrasonic beam. In this study we used gated magnetic resonance imaging (MRI) to investigate the inflow velocity in the LV transverse directions during early diastole in patients with DCM. METHODS: We studied 10 patients with DCM (mean age: 47 y). Ten age-matched healthy volunteers were also examined. Gradient echo images of the LV were obtained. Left ventricular short axis phase contrast images were obtained at the level of the mitral valve tip and 1 cm inside the LV. Long axis images were also obtained. Through-plane peak velocities at peak early diastolic filling were measured along the LV long axis, antero posterior short axis, and right-left short axis. Blood velocity was measured in 50 ms blocks. RESULTS: Early diastolic inflow velocity along the long axis, especially at the center of the LV, was well preserved in DCM. However, the inflow velocity in the antero-posterior transverse direction of the LV (i.e., in the direction of mitral valve excursion) was significantly reduced in DCM. CONCLUSIONS: Early diastolic inflow velocity in the antero-posterior transverse direction of the LV is reduced in patients with DCM indicating that the vector component of the forces acting in the antero-posterior transverse direction of the LV may be decreased in patients with DCM during early diastolic filling. PMID- 10832625 TI - Chylous ascites due to constrictive pericarditis. AB - Chylous ascites due to constrictive pericarditis is an extremely rare clinical entity, possibly caused by the augmented lymph production and high impedance to lymph drainage due to central venous hypertension. The authors describe a patient with chylous ascites caused by constrictive pericarditis in the absence of lymphatic obstruction. Cardiac catheterization is essential for the confirmation of accurate diagnosis of constrictive pericarditis. Magnetic resonance imaging of the heart is also very helpful in the diagnosis. The patient was symptom free and his ascites and edema completely resolved after pericardiectomy. PMID- 10832626 TI - An iso-integral mapping technique using magnetocardiogram, and its possible use for diagnosis of ischemic heart disease. AB - We have developed an iso-integral mapping technique that uses magneto-cardiogram (MCG) data to obtain a map as projected total current image on the torso from the heart. We have also investigated the applicability of iso-integral mapping to the diagnosis of ischemic heart disease. We simulated and measured the characteristics of two types of iso-integral maps: one using tangential (Bxy) components, and one using the normal component (Bz). Each vector component was measured by two types of superconducting quantum interference device (SQUID) system to determine the tangential and normal components. The tangential component of the magnetic field appeared to be equivalent to the current image in the myocardium projected on the observing plane, and we were able to obtain a projected total current image by integration of the tangential components during the depolarization and repolarization processes. And we found that the iso integral maps of normal hearts showed similar pattern in both processes; however, those of ischemic hearts showed different patterns. PMID- 10832627 TI - Primitive anaerobic protozoa: a false concept? PMID- 10832628 TI - Primitive anaerobic protozoa: the wrong host for mitochondria and hydrogenosomes? PMID- 10832629 TI - Replicative ageing and senescence in Saccharomyces cerevisiae and the impact on brewing fermentations. PMID- 10832630 TI - The Candida albicans antiporter gene CNH1 has a role in Na+ and H+ transport, salt tolerance, and morphogenesis. AB - The isolation and functional characterization of a Candida albicans Na+/H+ antiporter gene, CNH1, is reported here. The gene encodes a protein of 840 amino acids that exhibits high levels of similarity in sequence, size, and structural and functional domains to a group of known Na+/H+ antiporters of fungi. The CNH1 gene is able to functionally complement the salt-sensitivity of a Saccharomyces cerevisiae ena1 nha1 mutant, and mutations of two conserved aspartate residues to asparagines in the putative Na+-binding site abolish this activity. Deletion of CNH1 results in retardation of growth and a highly elongated morphology in a significant fraction of cells under conditions that normally support yeast growth. These results indicate that CNH1 has a role in Na+ and H+ transport, salt tolerance, and morphogenesis. PMID- 10832631 TI - Two distinct 18S rRNA secondary structures in Dipodascus (Hemiascomycetes). AB - The nucleotide sequences of the 18S rRNA gene from ascomycetous yeast-like fungi in the genera Dipodascus, Galactomyces and Geotrichum were determined and the tested strains were separated into two groups by sequence length. In group 1, the length and secondary structure of 18S rRNA corresponded to those of typical eukaryotes. In group 2, the 18S rRNA gene sequences were about 150 nt shorter than those of most other eukaryotes and the predicted secondary structure lacked helices 10 and E21-5. Many substitutions and some deletions in group 2 18S rRNA gene were not only found in variable regions, but also in regions that are highly conserved among ascomycetes. Despite the considerable differences in 18S rRNA gene sequence and secondary structure between group 2 and other fungi, including group 1, phylogenetic analysis revealed that groups 1 and 2 are closely related. These findings suggest that a number of deletions occurred in the 18S rRNA of the common ancestor of group 2 strains. PMID- 10832632 TI - The exbD2 gene as well as the iron-uptake genes tonB, exbB and exbD1 of Xanthomonas campestris pv. campestris are essential for the induction of a hypersensitive response on pepper (Capsicum annuum). AB - The tonB, exbB and exbD1 genes of Xanthomonas campestris pv. campestris are essential for ferric iron uptake. In contrast, the exbD2 gene located in the same gene cluster is not essential. Mutational analysis revealed that the ferric-iron uptake genes tonB, exbB and exbD1 are necessary for the induction of a hypersensitive response (HR) on the nonhost plant pepper (Capsicum annuum) and the induction of typical black rot symptoms on the host plant cauliflower (Brassica oleracea). Again, the exbD2 gene behaved differently. It was found to play a role only in the induction of the HR in pepper but not in the induction of black rot symptoms in cauliflower. Due to the low iron concentration in the plant tissue, the titre of viable bacteria of the ferric-iron-uptake mutants tonB, exbB and exbD1 decreased after leaf infiltration of pepper. The exbD2 mutant, however, which is not impaired in ferric iron uptake, multiplied in the pepper leaf tissue and grew even better than the wild-type strain, probably due to its failure to induce the HR. Nevertheless, the tonB, exbB and exbD1 mutant strains were able to spread systemically in cauliflower. PMID- 10832633 TI - 2-Oxoacid dehydrogenase multienzyme complexes in the halophilic Archaea? Gene sequences and protein structural predictions. AB - All Archaea catalyse the conversion of pyruvate to acetyl-CoA via a simple pyruvate oxidoreductase. This is in contrast to the Eukarya and most aerobic bacteria, which use the pyruvate dehydrogenase multienzyme complex [PDHC], consisting of multiple copies of three component enzymes: E1 (pyruvate decarboxylase), E2 (lipoate acetyl-transferase) and E3 (dihydrolipoamide dehydrogenase, DHLipDH). Until now no PDHC activity has been found in the Archaea, although DHLipDH has been discovered in the extremely halophilic Archaea and its gene sequence has been determined. In this paper, the discovery and sequencing of an operon containing the DHLipDH gene in the halophilic archaeon Haloferax volcanii are reported. Upstream of the DHLipDH gene are 3 ORFs which show highest sequence identities with the E1alpha, E1beta and E2 genes of the PDHC from gram-positive organisms. Structural predictions of the proposed protein product of the E2 gene show a domain structure characteristic of the E2 component in PDHCs, and catalytically important residues, including the lysine to which the lipoic acid cofactor is covalently bound, are conserved. Northern analyses indicate the transcription of the whole operon, but no PDHC enzymic activity could be detected in cell extracts. The presence in the E2 gene of an insertion (equivalent to approximately 100 aa) not found in bacterial or eukaryal E2 proteins, might be predicted to prevent multienzyme complex assembly. This is the first detailed report of the genes for a putative 2-oxoacid dehydrogenase complex in the Archaea, and the evolutionary and metabolic consequences of these findings are discussed. PMID- 10832634 TI - Era GTPase of Escherichia coli: binding to 16S rRNA and modulation of GTPase activity by RNA and carbohydrates. AB - Era, an essential GTPase, appears to play an important role in the regulation of the cell cycle and protein synthesis of bacteria and mycoplasmas. In this study, native Era, His-tagged Era (His-Era) and glutathione S-transferase (GST)-fusion Era (GST-Era) proteins from Escherichia coli were expressed and purified. It was shown that the GST-Era and His-Era proteins purified by 1-step affinity column chromatographic methods were associated with RNA and exhibited a higher GTPase activity. However, the native Era protein purified by a 3-step column chromatographic method had a much lower GTPase activity and was not associated with RNA which had been removed during purification. Purified GST-Era protein was shown to be present as a high- and a low-molecular-mass forms. The high-molecular mass form of GST-Era was associated with RNA and exhibited a much higher GTPase activity. Removal of the RNA associated with GST-Era resulted in a significant reduction in the GTPase activity. The RNA associated with GST-Era was shown to be primarily 16S rRNA. A purified native Era protein preparation, when mixed with total cellular RNA, was found to bind to some of the RNA. The native Era protein isolated directly from the cells of a wild-type E. coli strain was also present as a high-molecular-mass form complexed with RNA and RNase treatment converted the high-molecular-mass form into a 32 kDa low-molecular-mass form, a monomer of Era. Furthermore, a C-terminally truncated Era protein, when expressed in E. coli, did not bind RNA. Finally, the GTPase activity of the Era protein free of RNA, but not the Era protein associated with the RNA, was stimulated by acetate and 3-phosphoglycerate. These carbohydrates, however, failed to activate the GTPase activity of the C-terminally truncated Era protein. Thus, the results of this study establish that the C-terminus of Era is essential for the RNA-binding activity and that the RNA and carbohydrates modulate the GTPase activity of Era possibly through a similar mechanism. PMID- 10832635 TI - Antibacterials that are used as growth promoters in animal husbandry can affect the release of Shiga-toxin-2-converting bacteriophages and Shiga toxin 2 from Escherichia coli strains. AB - Antibiotics are commonly used as growth promoters in animal husbandry worldwide. This practice has been linked to the emergence of particular antibiotic-resistant bacteria, and is now controversial. In this study, the ability of growth promoting antibiotics to induce Shiga toxin (Stx)-converting bacteriophages from Stx-producing Escherichia coli (STEC) strains was investigated. Subinhibitory concentrations of the antibacterial growth promoters olaquindox, carbadox, tylosin and monensin were used for induction experiments. The amount of mature Stx-converting phage particles released from induced and non-induced cultures was determined, and the production of Stx was simultaneously measured by ELISA. Whereas the quinoxaline-1,4-dioxide-type antibiotics olaquindox and carbadox enhanced the release of Stx-converting phage particles from STEC cells, tylosin and monensin decreased phage induction. The production of Stx increased or decreased simultaneously with the amount of free phages. The results of this study show that particular antibacterial growth promoters can induce Stx phages. In vivo induction of Stx phages from lysogenic STEC may increase the amount of free phages in the intestine and therefore may contribute to the spread of STEC and development of new STEC pathotypes. PMID- 10832636 TI - High morpholine degradation rates and formation of cytochrome P450 during growth on different cyclic amines by newly isolated Mycobacterium sp. strain HE5. AB - Using morpholine as sole source of carbon, nitrogen and energy, strain HE5 (DSM 44238) was isolated from forest soil. The isolated strain was identified as a member of the subgroup of fast-growing Mycobacterium species as revealed by 16S rDNA analysis. An identity of 99.4% was obtained to Mycobacterium gilvum; however, the type strain was unable to utilize morpholine. A maximal growth rate of 0.17 h(-1) was observed at a morpholine concentration of 30 mM, 30 degrees C and pH 7.2. The substrate was tolerated at concentrations up to 100 mM. Besides morpholine, the strain utilized pyrrolidine, piperidine and proposed intermediates in morpholine metabolism such as glycolate, glyoxylate and ethanolamine. Degradation of morpholine, piperidine and pyrrolidine by resting or permeabilized cells was strictly dependent on the presence of oxygen. Addition of the cytochrome-P450-specific inhibitor metyrapone to the growth medium resulted in a significantly decreased growth rate if these cyclic amines were used as a substrate. Carbon monoxide difference spectra of crude extracts from cells grown on these substrates compared to spectra obtained for extracts of succinate-grown cells indicated that cytochrome P450 is specifically expressed during growth on the cyclic amines. These data indicated that a cytochrome-P450-dependent monooxygenase is involved in the degradation of the three cyclic amines. PMID- 10832637 TI - Microbial community changes in biological phosphate-removal systems on altering sludge phosphorus content. AB - Biomarkers (respiratory quinones and cellular fatty acids) and denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S rRNA genes were used to characterize the microbial community structure of lab-scale enhanced biological phosphate-removal (EBPR) systems in response to altering sludge phosphorus (P) content. All the data suggest that the microbial community structures of sludge samples with a P content between 8 and 12.3% (sludge dry weight) (i.e. good EBPR activity) were very similar, but differed from those with 2% P content (i.e. no EBPR activity). For all samples analysed, ubiquinones Q-8 and Q-10, menaquinone MK-8(H4), and fatty acids C16:0, C16:1 omega9c and C18:1, omega11c were the major components. The dominance of Q-8, Q-10 and MK-8(H4) suggested that large numbers of organisms belonging to the beta and alpha subclasses of the Proteobacteria and the Actinobacteria from the high G+C Gram-positive bacteria, respectively, were present. DGGE analysis revealed at least 7-9 predominant DNA bands and numerous other fragments in each sample. Five major DGGE fragments from each of the 2% and 12% P-containing sludge samples, respectively, were successfully isolated and sequenced. Phylogenetic analysis of the sequences indicated that both 2% and 12% P-containing sludge samples shared three common phylotypes that were separately affiliated with a novel bacterial group from the gamma subclass of the Proteobacteria, two MK-8(H4)-containing actinobacteria previously isolated from the 2% P-containing sludge, and a Caulobacter spp. in the alpha subclass of the Proteobacteria. The phylogenetic analysis also revealed phylotypes unique to both sludge samples. Changes in sludge P content therefore had an effect on the composition and abundance of the predominant microbial populations, though specific phylotypes could not be unequivocally associated with EBPR. PMID- 10832638 TI - Cadmium uptake and subcellular compartmentation in the ectomycorrhizal fungus Paxillus involutus. AB - Cadmium uptake and subcellular compartmentation in the ectomycorrhizal fungus Paxillus involutus were investigated using radiotracer flux analyses. Concentration-dependent Cd2+-uptake kinetics were characterized by a smooth, non saturating curve that could be dissected into linear and saturable components. The linear-uptake kinetic component was interpreted as representing binding of Cd to apoplastic components, whereas the remaining saturable component was the result of carrier-mediated transport across the plasma membrane. Cell-wall-bound Cd was almost completely removed during desorption from cell-wall preparations. Cd2+ desorption from intact mycelium was found to be a function of time involving three compartments corresponding in series to cell wall (50%), cytoplasm (30%) and vacuole (20%), when mycelia were exposed to a 0.05 microM Cd concentration. At 4 degrees C, most of the Cd recovered was due to the cell-wall-bound fraction, suggesting that transport across the plasma membrane is a metabolically mediated process. Carbonyl cyanide m-chlorophenylhydrazone (CCCP) inhibited Cd accumulation in P. involutus mycelia by up to 28%, which indicates that transport of Cd2+ was partially dependent on the membrane potential. Cd2+ uptake into symplasm is linked to Ca2+ transport, as revealed by the inhibition of Cd accumulation by the Ca2+ ionophore A23187. The present work demonstrates the ability of the ectomycorrhizal fungus P. involutus to take up and further accumulate Cd in different compartments. Binding of Cd onto cell walls and accumulation of Cd in the vacuolar compartment may be regarded as two essential metal-detoxification mechanisms. These data represent a first step towards the understanding of metal-tolerance mechanisms in mycorrhizal fungi. PMID- 10832639 TI - Ferritin from the obligate anaerobe Porphyromonas gingivalis: purification, gene cloning and mutant studies. AB - Porphyromonas gingivalis is an obligate anaerobe that utilizes haem, transferrin and haemoglobin efficiently as sources of iron for growth, and has the ability to store haem on its cell surface, resulting in black pigmentation of colonies on blood agar plates. However, little is known about intracellular iron storage in this organism. Ferritin is one of the intracellular iron-storage proteins and may also contribute to the protection of organisms against oxidative stresses generated by intracellular free iron. A ferritin-like protein was purified from P. gingivalis and the encoding gene (ftn) was cloned from chromosomal DNA using information on its amino-terminal amino acid sequence. Comparison of the amino acid sequence deduced from the nucleotide sequence of ftn with those of known ferritins and bacterioferritins identified the protein as a ferritin and positioned it between proteins from the Proteobacteria and Thermotogales. The P. gingivalis ferritin was found to contain non-haem iron, thus confirming its identity. Construction and characterization of a P. gingivalis ferritin-deficient mutant revealed that the ferritin was particularly important for the bacterium to survive under iron-depleted conditions (both haemin and transferrin starvation), indicating that intracellular iron is stored in ferritin regardless of the iron source and that the iron stored in ferritin is utilized under iron-restricted conditions. However, the ferritin appeared not to contribute to protection against oxidative stresses caused by peroxides and atmospheric oxygen. PMID- 10832640 TI - Metabolism of carveol and dihydrocarveol in Rhodococcus erythropolis DCL14. AB - Rhodococcus erythropolis DCL14 assimilates all stereoisomers of carveol and dihydrocarveol as sole source of carbon and energy. Induction experiments with carveol- or dihydrocarveol-grown cells showed high oxygen consumption rates with these two compounds and with carvone and dihydrocarvone. (Dihydro)carveol-grown cells of R. erythropolis DCL14 contained the following enzymic activities involved in the carveol and dihydrocarveol degradation pathways of this micro organism: (dihydro)carveol dehydrogenase (both NAD+- and dichlorophenolindophenol dependent activities), an unknown cofactor-dependent carvone reductase, (iso )dihydrocarvone isomerase activity, NADPH-dependent dihydrocarvone monooxygenase (Baeyer-Villiger monooxygenase), epsilon-lactone hydrolase and an NAD+-dependent 6-hydroxy-3-isopropenylheptanoate dehydrogenase. Product accumulation studies identified (4R)-carvone, (1R,4R)-dihydrocarvone, (4R,7R)-4-isopropenyl-7-methyl-2 oxo-oxepanone, (3R)-6-hydroxy-3-isopropenylheptanoate, (3R)-3-isopropenyl-6 oxoheptanoate, (3S,6R)-6-isopropenyl-3-methyl-2-oxooxepanone and (5R)-6-hydroxy-5 isopropenyl-2-methylhexanoate as intermediates in the (4R)-carveol degradation pathway. The opposite stereoisomers of these compounds were identified in the (4S)-carveol degradation pathway. With dihydrocarveol, the same intermediates are involved except that carvone was absent. These results show that R. erythropolis DCL14 metabolizes all four diastereomers of carveol via oxidation to carvone, which is subsequently stereospecifically reduced to (1R)-(iso-) dihydrocarvone. At this point also dihydrocarveol enters the pathway, and this compound is directly oxidized to (iso-)dihydrocarvone. Cell extracts contained both (1R)-(iso )dihydrocarvone 1,2-monooxygenase and (1S)-(iso)-dihydrocarvone 2,3-monooxygenase activity, resulting in a branch point of the degradation pathway; (1R)-(iso )dihydrocarvone was converted to 4-isopropenyl-7-methyl-2-oxo-oxepanone, while (1S)-(iso)-dihydrocarvone, which in vivo is isomerized to (1R)-(iso )dihydrocarvone, was converted to 6-isopropenyl-3-methyl-2-oxo-oxepanone. 4 Isopropenyl-7-methyl-2-oxooxepanone is hydrolysed to 6-hydroxy-3 isopropenylheptanoate, which is subsequently oxidized to 3-isopropenyl-6 oxoheptanoate, thereby linking the (dihydro)carveol degradation pathways to the limonene degradation pathway of this micro-organism. 6-Isopropenyl-3-methyl-2-oxo oxepanone is, in vitro, hydrolysed to 6-hydroxy-5-isopropenyl-2-methylhexanoate, which is thought to be a dead-end metabolite. PMID- 10832641 TI - Rhodococcus opacus strain PD630 as a new source of high-value single-cell oil? Isolation and characterization of triacylglycerols and other storage lipids. AB - The triacylglycerol (TAG)-accumulating, hydrocarbon-degrading bacterium Rhodococcus opacus strain PD630 and chemically induced storage-deficient mutants derived from this strain were investigated for their capability to accumulate storage lipids in the cytoplasm during cultivation under nitrogen-limiting conditions. Acylglycerols were analysed by matrix-associated laser desorption ionization-time of flight mass spectrometry (MALDI-TOF) and by reversed-phase HPLC. Fatty acids comprising 13-19 carbon atoms in various acylglycerols constituted up to 76% of the cellular dry weight in gluconate-grown cells, with a significant proportion of odd-numbered fatty acids. Hydrolysis using pancreatic lipase and deacylation with ethyl magnesium bromide were employed to identify the stereospecific distribution of fatty acids at the glycerol. This analysis showed that the fatty acids were not randomly distributed between the three positions of the glycerol backbone. In comparison with common plant fats, where the longer and higher unsaturated fatty acids are predominantly found at position 2, R. opacus PD630 accumulated only the shorter and saturated fatty acids in this position. More than 100 mutants accumulating TAG at a significantly lower rate were obtained by chemical mutagenesis and identified by staining with Sudan Black B. All the mutants showed similar neutral lipid patterns by TLC analysis, with a small distinct spot exhibiting the same R(F) value as TAG; this was identified as a residual amount of TAG by preparative TLC and MALDI-TOF, indicating that this bacterium is possibly capable of synthesizing TAGs by at least two different pathways. PMID- 10832642 TI - Additive effects of a two-amino-acid insertion and a single-amino-acid substitution in dihydropteroate synthase for the development of sulphonamide resistant Neisseria meningitidis. AB - Sulphonamide resistance in some clinical isolates of Neisseria meningitidis is associated with an insertion in the chromosomal folP gene leading to the addition of two amino acids, serine and glycine, in the drug target enzyme dihydropteroate synthase (DHPS). Removal of the insertion resulted in a markedly higher Km for the substrate p-aminobenzoic acid and a markedly lower Km for 2-amino-4-hydroxy-6 (hydroxymethyl)-7,8-dihydropteridine pyrophosphate. In the same isolates an additional important difference, compared to wild-type enzymes, was found at amino acid position 68, which is a proline in most DHPS enzymes, but is serine in one and leucine in another clinical isolate of sulphonamide-resistant N. meningitidis. The alteration at position 68 was found to affect mainly the level of sulphonamide resistance and had only a minor effect on the Km for the substrates. Introduction of the serine-glycine dipeptide at position 194 and a proline to serine substitution at position 68 in DHPS from normal, susceptible N. meningitidis failed to produce a functional sulphonamide-resistant enzyme. The conclusion of this study is that it is not possible to change a normal chromosomally encoded DHPS of N. meningitidis to a sulphonamide-resistant one simply by an insertion of serine and glycine as seen in clinical isolates. It is likely that the resistance gene found in clinical isolates has evolved in another bacterial species where a combination of other amino acid changes may have contributed to produce a functionally resistant enzyme. This new resistance gene may have then been introduced into N. meningitidis by natural transformation. PMID- 10832643 TI - Isolation of a novel insertion sequence from Mycobacterium fortuitum using a trap vector based on inactivation of a lacZ reporter gene. AB - An insertion sequence of Mycobacterium fortuitum has been isolated using a trap vector following insertion in and inactivation of the lacZ reporter gene. The trap vector is a temperature-sensitive (ts) Escherichia coli-mycobacterium shuttle plasmid, pCD4, which contains ts oriM, the kanamycin-resistance gene as a selection marker and a lacZ expression cassette. The ts mutation present in pCD4 functions in mycobacteria and enables screening for transposable elements from the mycobacterial genome that disrupt the lacZ gene by screening for white colonies on X-Gal plates in both mycobacterial as well as E. coli hosts. The vector was used to isolate a novel 1.653 kb insertion sequence from M. fortuitum named IS219. IS219 duplicated host DNA at the target site, had inverted repeats at its ends and contained two ORFs on one strand. One of the predicted proteins showed homology to a putative transposase from Acetobacter pasteurianus. IS219 was present in two copies in the genome of M. fortuitum. The trap vector appears to be useful in trapping insertion sequences from different mycobacteria by screening for the disrupted LacZ phenotype. PMID- 10832644 TI - Transposition in Lactobacillus sakei: inactivation of a second lactocin S operon by the insertion of IS1520, a new member of the IS3 family of insertion sequences. AB - The analysis of spontaneous bacteriocin-negative mutants has led to the identification and characterization of a new, transpositionally active, insertion sequence of the IS3 family in the lactocin-S-producing Lactobacillus sakei strain L45. The element, which has been designated IS1520, is 1302 bp long with 10 bp perfect inverted repeat ends and generates direct repeats of a trinucleotide of target sequence upon transposition to the lactocin S locus. IS1520 encodes two consecutive, partially overlapping, major ORFs, which are frameshifted in a manner typical of the IS3 family. Despite a low overall DNA sequence similarity, the putative frameshifting region of IS1520 is highly similar to the corresponding region of IS1163, which is a related element previously shown to be active in L. sakei L45. PMID- 10832645 TI - The Salmonella FlgA protein, a putativeve periplasmic chaperone essential for flagellar P ring formation. AB - P ring is a periplasmic substructure of the flagellar basal body and is believed to connect with the peptidoglycan layer in Salmonella. Two flagellar genes, flgA and flgI, are known to be indispensable for P ring formation. The flgI gene encodes the component protein of the P ring. However, the role of the flgA gene product in P ring assembly remained unknown. Here, evidence is presented that FlgA is synthesized as a precursor form and exported via the Sec secretory pathway into the periplasmic space where P ring formation takes place. Overproduction of the FlgI protein led flgA mutants to form flagella with a P ring, suggesting that FlgA plays an auxiliary role in P ring assembly. Far Western blot analysis revealed that FlgA binds in vitro to both FlgI and FlgA itself. Though a direct FlgI-FlgI interaction in the absence of FlgA could not be demonstrated, an indirect or direct interaction between the FlgI proteins was observed in the presence of FlgA. FlgA alone was very unstable in vivo, but co expression with FlgI could stabilize FlgA. This suggests the presence of FlgA FlgI interaction in vivo. On the basis of these results, a hypothesis is proposed that FlgA acts as a periplasmic chaperone, which assists a polymerization reaction of FlgI into the P ring through FlgA-FlgI interaction. PMID- 10832646 TI - Inactivation of the ptsI gene encoding enzyme I of the sugar phosphotransferase system of Streptococcus salivarius: effects on growth and urease expression. AB - The urease genes of Streptococcus salivarius 57.1 are tightly repressed in cells growing at neutral pH. When cells are cultivated at acidic pH values, the urease genes become derepressed and transcription is enhanced when cells are growing under carbohydrate-excess conditions. Previously, the authors proposed that the bacterial sugar:phosphotransferase system (PTS) modulated the DNA-binding activity by phosphorylation of the urease repressor when carbohydrate was limiting. The purpose of this study was to assess whether enzyme I (EI) of the PTS could be involved in modulating urease expression in response to carbohydrate availability. An EI-deficient strain (ptsI18-3) of S. salivarius 57.1 was constructed by insertional inactivation of the ptsI gene. The mutant had no measurable PTS activity and lacked EI, as assessed by Western analysis. The mutant grew as well as the wild-type strain on the non-PTS sugar lactose, and grew better than the parent when another non-PTS sugar, galactose, was the sole carbohydrate. The mutant was able to grow with glucose as the sole carbohydrate, but displayed a 24 h lag time and had a generation time some threefold longer than strain 57.1. The mean OD600 attained after 48 h by ptsI18-3 supplied with fructose was 0.16, with no additional growth observed even after 3 d. Urease expression in the wild-type and mutant strains was assessed in continuous chemostat culture. Repression of urease at neutral pH was seen in both strains under all conditions tested. Growth of wild-type cells on limiting concentrations of lactose resulted in very low levels of urease expression compared with growth on PTS sugars. In contrast, under similar conditions, urease expression in ptsI18 3 was restored to levels seen in the parent growing on PTS sugars. Growth under conditions of lactose excess resulted in further derepression of urease, but ptsI18-3 expressed about threefold higher urease activity than 57.1. The results support a role for EI in urease regulation, but also indicate that additional factors may be important in regulating urease gene expression. PMID- 10832647 TI - Localization and characterization of the ligand-binding domain of the fibrinogen binding protein (FgBP) of Streptococcus equi subsp. equi. AB - The group C streptococcus Streptococcus equi subsp. equi possesses a 498-residue major cell-wall-associated protein (FgBP) which binds horse fibrinogen (Fg), reacts with convalescent horse serum and protects against lethal S. equi challenge in a small animal model. In the present study, analysis of a panel of 17 purified N- and C-terminal FgBP truncates by ligand affinity blotting and SDS PAGE revealed that the region required for maximum binding of Fg extended over the first half of the mature protein. The C-terminal two-thirds of this domain is predicted to be alpha-helical coiled-coil and the N-terminal one-third to possess non-coiled-coil single strands. Residues at the extreme N-terminus and within the coiled-coil region are both required for ligand binding. A high incidence of alpha-helical coiled-coil structure also seems to be responsible in part for the aberrant mobility of FgBP on SDS gels. The efficiency with which FgBP binds Fg from different animal species decreases in the order horse > mouse, pig > rat > sheep, dog, bovine, human. Binding to horse Fg is inversely related to temperature over the range 45-4 degrees C and is independent of Ca2+ ions. MS analysis provided corroborative evidence that FgBP is covalently linked to the cell wall peptidoglycan. PMID- 10832648 TI - emm and sof gene sequence variation in relation to serological typing of opacity factor-positive group A streptococci. AB - Approximately 40-60% of group A streptococcal (GAS) isolates are capable of opacifying sera, due to the expression of the sof (serum opacity factor) gene. The emm (M protein gene) and sof 5' sequences were obtained from a diverse set of GAS reference strains and clinical isolates, and correlated with M serotyping and anti-opacity-factor testing results. Attempts to amplify sof from strains with M serotypes or emm types historically associated with the opacity-factor-negative phenotype were negative, except for emm12 strains, which were found to contain a highly conserved sof sequence. There was a strong correlation of certain M serotypes with specific emm sequences regardless of strain background, and likewise a strong association of specific anti-opacity-factor (AOF) types to sof gene sequence types. In several examples, M type identity, or partial identity shared between strains with differing emm types, was correlated with short, highly conserved 5' emm sequences likely to encode M-type-specific epitopes. Additionally, each of three pairs of historically distinct M type reference strains found to share the same 5' emm sequence, were also found to share M serotype specificity. Based upon sof sequence comparisons between strains of the same and of differing AOF types, an approximately 450 residue domain was determined likely to contain key epitopes required for AOF type specificity. Analysis of two Sof sequences that were not highly homologous, yet shared a common AOF type, further implicated a 107 aa portion of this 450-residue domain in putatively containing AOF-specific epitopes. Taken together, the serological data suggest that AOF-specific epitopes for all Sof proteins may reside within a region corresponding to this 107-residue sequence. The presence of specific, hypervariable emm/sof pairs within multiple isolates appears likely to be a reliable indicator of their overall genetic relatedness, and to be very useful for accurate subtyping of GAS isolates by an approach that has relevance to decades of past M-type-based epidemiological data. PMID- 10832649 TI - Molecular characterization of Bordetella bronchiseptica filamentous haemagglutinin and its secretion machinery. AB - Two closely related pathogens, Bordetella pertussis and Bordetella bronchiseptica, share a number of virulence factors. Filamentous haemagglutinin (FHA) is widely regarded as the dominant adhesin of B. pertussis, and its multiple binding activities have been well characterized. This large protein is produced and secreted at high levels by B. pertussis and significantly lower levels by B. bronchiseptica strains. FHA secretion is mediated by a single outer membrane accessory protein, FhaC. The genes encoding FHA and FhaC in B. bronchiseptica were characterized by sequencing and functional analyses and are highly similar to those of B. pertussis. The most distinctive feature of B. bronchiseptica FHA is additional repeats in the N-terminal portion of the predicted protein. Interestingly, a point mutation in the fhaB promoter region of the B. bronchiseptica GP1 isolate, relative to other isolates, was found to be detrimental to promoter activity and to FHA production. FhaC and the N-terminal secretion domain of FHA of B. bronchiseptica were fully functional for secretion in B. pertussis. Thus, the different levels of FHA secretion by these Bordetella species might reflect differences in physiology, composition and structure of cell envelope, or differential protein degradation. Characterization of FHA expression and function may provide clues as to the basis of host species tropism, tissue localization and receptor recognition. PMID- 10832650 TI - Lymphocytic infiltration in the chicken trachea in response to Mycoplasma gallisepticum infection. AB - A prominent feature of disease induced by Mycoplasma gallisepticum is a lymphoproliferative response in the respiratory tract. Although this is also seen in other mycoplasma infections, including Mycoplasma pneumoniae, the phenotype of the lymphocytes infiltrating the respiratory tract has not been determined. In this study, the numbers and distribution of lymphocytes in the tracheas of chickens infected with a virulent strain of M. gallisepticum were examined. Three groups of chickens were experimentally infected with M. gallisepticum and three unchallenged groups were used as controls. One infected and one control group were culled at 1, 2 and 3 weeks post infection. Tracheas were removed and examined for the presence and number of T cells carrying CD4, CD8, TCRgamma7, TCRalphabeta1 or TCRalphabeta2 markers. There was no significant difference in the number of CD8+ cells in the upper, middle and lower trachea. High numbers of both CD4+ and CD8+ cells were found with variable numbers of TCRalphabeta1+ and TCRalphabeta2+, but no TCRgammadelta+, cells throughout the time course. The distribution of CD4 cells was dispersed, while the CD8+ cells were clustered in follicular-like arrangements. No difference was detected in the distribution of TCRalphabeta1+ and TCRalphabeta2+ cells. The titre of mycoplasma genomes in the trachea decreased significantly from 1 to 2 weeks, while the mucosal thickness of the trachea increased significantly from 1 to 2 weeks then decreased from 2 to 3 weeks, indicating resolution of the lesions following control of infection. This study is the first to examine the phenotypes of T lymphocytes infiltrating the respiratory tract during mycoplasma infections. The findings suggest involvement of specific stimulation of CD8+ cells, particularly in the acute phase of disease. PMID- 10832651 TI - Neochlamydia hartmannellae gen. nov., sp. nov. (Parachlamydiaceae), an endoparasite of the amoeba Hartmannella vermiformis. AB - Free-living amoebae are increasingly being recognized to serve as vehicles of dispersal for various bacterial human pathogens and as hosts for a variety of obligate bacterial endocytobionts. Several Chlamydia-like Acanthamoeba endocytobionts constituting the recently proposed family Parachlamydiaceae are of special interest as potential human pathogens. In this study coccoid bacterial endocytobionts of a Hartmannella vermiformis isolate were analysed. Infection of H. vermiformis with these bacteria resulted in prevention of cyst formation and subsequent host-cell lysis. Transfection experiments demonstrated that the parasites were not capable of propagating within other closely related free living amoebae but were able to infect the distantly related species Dictyostelium discoideum. Electron microscopy of the parasites revealed typical morphological characteristics of the Chlamydiales, including the existence of a Chlamydia-like life-cycle, but indicated that these endocytobionts, in contrast to Chlamydia species, do not reside within a vacuole. Comparative 16S rRNA sequence analysis showed that the endocytobiont of H. vermiformis, classified as Neochlamydia hartmannellae gen. nov., sp. nov., is affiliated to the family Parachlamydiaceae. Confocal laser scanning microscopy in combination with fluorescence in situ hybridization using rRNA-targeted oligonucleotide probes confirmed the intracellular localization of the parasites and demonstrated the absence of other bacterial species within the Hartmannella host. These findings extend our knowledge of the phylogenetic diversity of the Parachlamydiaceae and demonstrate for the first time that these endocytobionts can naturally develop within amoebae of the genus Hartmannella. PMID- 10832653 TI - The optimal method of coronary revascularization in dialysis patients: choosing between a rock and a hard place. PMID- 10832652 TI - Identification of two genetic groups in Bacteroides fragilis by multilocus enzyme electrophoresis: distribution of antibiotic resistance (cfiA, cepA) and enterotoxin (bft) encoding genes. AB - Ninety-three Bacteroides fragilis strains of different origin were analysed by multilocus enzyme electrophoresis (MLEE). Fourteen of the 15 genetic loci analysed were polymorphic, whilst nucleoside phosphorylase was monomorphic. There was a mean of six alleles per locus and a mean genetic diversity of 0.393. Cluster analysis identified 90 electrophoretic types (ETs) separated into two major phylogenetic divisions at a genetic distance of 0.70. Division I consisted of 81 ETs carrying the endogenous class A beta-lactamase gene cepA, whereas division II comprised 9 ETs carrying the class B beta-lactamase gene cfiA, but not cepA. The presence of these two genes was assessed by PCR and the expression of the cfiA gene was investigated by determining the level of resistance to the antibiotic imipenem. MLEE showed a smaller genetic distance among the genotypes of the imipenem-resistant than among the imipenem-susceptible strains. No other particular cluster was observed. The enterotoxin gene (bft) was detected by PCR: DNA sequencing of the products obtained showed that the different bft alleles (bft-1, bft-2 and bft-3) were scattered randomly troughout the phylogenetic tree. No association between distinct clones and clinical manifestations (sepsis, abscesses, diarrhoea), geographical origin or host origin (human or animal) could be found. PMID- 10832654 TI - Closed stiff-wire manipulation of malpositioned Tenckhoff catheters offers a safe and effective way of prolonging peritoneal dialysis. PMID- 10832655 TI - New approach to preoperative autologous blood donation (PABD) PMID- 10832656 TI - Direct infusion of ascites into the blood circuit during hemodiafiltration in uremic patients with cirrhosis. AB - Two chronic dialysis patients with massive ascites caused by cirrhosis were treated by infusion of their ascites directly into the blood circuit. This stabilized their hemodynamics during dialysis, facilitating the control of weight gain and ascites, and thus markedly improving their general condition. Long-term use of this therapy was able to prevent the accumulation of ascitic fluid. Interestingly, fever occurred when this therapy was performed with hemodialysis, but not with hemofiltration or hemodiafiltration, suggesting that a pyrogen in the ascites was removed by filtration. PMID- 10832657 TI - Sleep disorders in peritoneal and haemodialysis patients as assessed by a self administered questionnaire. AB - Sleep disorders have been reported as a frequent problem in dialysis patients. However, only one paper has compared the prevalence and possible causes of this complication in peritoneal (PD) and haemodialysis (HD) patients. We surveyed 84 PD and 87 HD patients about disordered sleep using a self-administered questionnaire. Forty-nine percent of PD and 56% of HD patients reported problems sleeping. These problems were rated as severe by 29 PD and 22 HD patients. Type of disturbances involved delayed sleeping (13 PD and 32 HD, p < 0.005), interrupted sleep (32 PD and 44 HD) and early morning awakening (25 PD and 37 HD). The number of hours of sleep varied widely among patients: it was 5 and 21 minutes in PD patients with sleep disorders and 7 and 37 min in PD pts without such problems. No statistically significant relationship was evidenced between sleep disorders and age, sex, body weight, obesity, duration of dialysis, dialysis dose, self-assessed sadness, anxiety, worry, pain, pruritus, dyspnoea, restless leg syndrome, use of cigarettes, caffeine, or sleeping pills. In conclusion, sleep disorders are a frequent problem in both PD and HD patients. Apparently the relationship with demographics, dialysis dose, lifestyle and personality traits is poor. The possible role of other causes should be investigated. PMID- 10832658 TI - Effects of anion exchange resin as phosphate binder on serum phosphate and iPTH levels in normal rats. AB - In order to investigate the characteristics of anion exchange resins that may safely and effectively bind dietary phosphate in digestive tract, phosphate binding experiments were carried out in vitro and in vivo with normal rats by comparing anion exchange resins, PAA-B (which has the same chemical structure as Sevelamer HCl) and Dowe 1x8, with CaCO3. In in vitro phosphate binding experiments, PAA-B bound 32.3% less phosphate than CaCO3 at pH 7. In the rat dietary phosphorus excretion experiments, PAA-B, Dowex 1x8, and CaCO3 increased fecal phosphorus excretion by 62.7, 32.3, and 84.0%, respectively. Famotidine significantly reduced the phosphate binding of CaCO3. When phosphate solution was orally administered, PAA-B depressed serum phosphorus augmentations immediately after administration and thereafter effectively depressed serum iPTH. This suggests that anion exchange resins with most primary and secondary amino type anion exchange groups, have bright prospects in the treatment of hyperphosphatemia. PMID- 10832659 TI - Heparin coating of extracorporeal circuits inhibits cytokine release from mononuclear cells during cardiac operations. AB - PURPOSE: To evaluate whether the production of interleukin 2 (IL 2), interleukin 6 (IL 6) and interleukin 10 (IL 10) from stimulated peripheral blood mononuclear cells (PBMC) was affected by coating extracorporeal circuits in patients undergoing cardiopulmonary bypass (CPB). In addition, postoperative clinical parameters were compared between patients with heparin-coated and uncoated CPB. DESIGN: Prospective, controlled in vivo/ex vivo study. PROCEDURE: Blood samples were drawn immediately before, at the end and 24 hours after the end of CPB using either a conventional circuit (n=10) or a heparin-coated circuit (n=10) in patients undergoing CPB. Cytokine release on the supernatants of activated PBMC was detected. Cardiopulmonary parameters were measured before CPB, at ICU admission, 3 hours and 24 hours after ICU admission in both groups of patients. Statistical difference intragroups and between groups were investigated with the analysis of variance for repeated measures. RESULTS: IL 6 and IL 10 release was significantly less (p<0.05) in the heparin-coated group. No differences in clinical parameters were observed between the two groups. CONCLUSIONS: These results suggest that with the use of heparin-coated circuits there is a lower production of IL 6 and IL 10 from isolated PBMC than with uncoated circuits. PMID- 10832660 TI - Alumina femoral head fracture: an in vitro study. AB - A fracture of a ceramic femoral head is reported in this study. Fractures of ceramic femoral heads are uncommon and reports on this complication are rare. After 3 million cycles, on a twelve station hip simulator that tested alumina femoral head against polyethylene acetabular cup, fracture of the ceramic ball was observed. The retrieved specimen consisted of three large ceramic fragments from the same ceramic femoral head, a polyethylene acetabular cup and a stainless steel jig. Careful and detailed examination of the removed components was made. The fracture of the ceramic ball resulted in damage to the metal taper of the jig component which was fixed into the simulator. PMID- 10832661 TI - Development of new tracheal prosthesis: autogenous mucosa-lined prosthesis made from polypropylene mesh. AB - Reliable tracheal or tissue graft has not been developed yet for the reconstruction of large, circumferential tracheal defects. Major limitations were anastomotic dehishence and stenosis, which were attributed to the poor epithelisation of the prosthetic graft. We developed a new tracheal prosthesis that has a viable lined and well-vascularized mucosa. The prosthesis consists of Prolene mesh reinforced with polypropylene rings, and is coated with gelatin. In addition, we lined the luminal surface of the prosthesis with transplanted autogenous oral mucosa and wrapped the prosthesis with greater omentum. Animal experiments were performed using 10 adult mongrel dogs. The transplanted mucosa and wrapped greater omentum tightly adhered to the prosthesis to make a single unit within two weeks. The mucosa survived well, was well vascularised by new vessels from greater omentum and showed normal histology. Complete surgical resection and replacement of a thoracic trachea (3 cm in length, 6 tracheal rings) were carried out in 2 dogs, which survived well with normal activity. We concluded that this highly biocompatible tracheal prosthesis could be very useful for step-wise reconstruction of tracheal defects. PMID- 10832662 TI - Development of a special catheterbag to enable artificial organ evaluation in conscious, unrestrained pigs: technical note. AB - Pigs are widely used as models for a variety of human diseases, because many of their physiological functions closely resemble those of humans. However, information on instrumentation techniques is still scarce. In particular, experiments in conscious pigs focused on extracorporeal circuits are connected to a variety of methodical problems with respect to the handling of the animals. Usually, pigs are placed in restraint-slings during the application of an extracorporeal system. However, this method of restraint may lead to excessive mental distress even in trained animals. The latter might influence the results and certainly affects principles of animal welfare. Our own experiences with instrumented, conscious, but unrestrained dogs encouraged us to modify methods used for the fixation of in-dwelling central venous catheters in dogs with special regard to the species specific behaviour and phenotype of pigs. A cord retractable leash (CRL) was used for maintaining a safe distance between the animal and the outer ends of the catheters. To prevent dehiscences of the required fixation sutures a new catheter bag (CB) was designed to counteract tension forces caused by the CRL's spring-mechanism. The combination of both the CRL and CB enabled us to conduct safe experiments with conscious, unrestrained pigs. We alleviated the mental distress these animals were exposed to in comparison to former methods based on restraint of the animals. PMID- 10832663 TI - The ovariectomised sheep as a model for testing biomaterials and prosthetic devices in osteopenic bone: a preliminary study on iliac crest biopsies. AB - A histomorphometric and ultrastructural evaluation on sheep iliac bone was performed. Six sheep were ovariectomised (OVX Group) and 6 were left intact (Sham aged, Control Group). An iliac crest biopsy was performed randomly in 6 animals at the beginning of the study, then, in all the animals, after 12 and 24 months. A significant decrease in trabecular bone volume, trabecular thickness (p<0.0005) and cell volume (p<0.005) was observed in OVX animals. A modest decrease in trabecular number and osteoid thickness together with an increase in trabecular separation were observed in OVX animals at 12 and 24 months. The osteoid volume showed a significant difference (p<0.05) between the groups. In OVX animals, at 12 months, Scanning Electron Microscopy revealed an enlargement of the trabecular space and a progressive replacement of bone matrix with adipose tissue. These signs were accentuated at 24 months. In conclusion, OVX sheep showed a loss of trabecular bone starting at 12 months after ovariectomy. The developed osteopenic state may be considered as a useful tool when doing research on biomaterial osteointegration. PMID- 10832664 TI - Evidence-based nephrology: efficacy of hepatitis B vaccine in hemodialysis patients. PMID- 10832665 TI - Diagnosis and non-surgical treatment of bile duct carcinoma: developments in the past decade. AB - In the past decade, ultrasonography and magnetic resonance cholangiography have become useful modalities for the screening of bile duct cancer. Intraductal ultrasonography, using a thin-caliber (2.0 to 2.4 mm in diameter) and high frequency (15 to 20 MHz) probe, has become a promising modality in assessing locoregional staging. For interventional therapy of unresectable bile duct carcinoma, metallic stents also became an excellent modality. However, tumor ingrowth and overgrowth into the mesh of stents are problems to be solved, and external radiation therapy is widely conducted to control the tumor. To improve the limitations of external radiation, brachytherapy, photodynamic therapy, and microwave coagulation therapy are conducted. However, these modalities are available in only a limited number of institutions even now. To predict the prognosis of the patients, approaches using molecular biology must be established. PMID- 10832666 TI - Endoscopic ultrasonography for diagnosis of submucosal invasion in early gastric cancer. AB - Endoscopic ultrasonography (EUS) is considered to be useful for deciding the treatment course for early gastric cancer. To determine reliable indications suggesting submucosal tumor invasion, we retrospectively analyzed EUS images of the hyperechoic third layer, which corresponds to the submucosa. The subjects enrolled in this study were 75 patients, with 78 gastric cancers (diagnosed as mucosal cancer without ulcerous changes on endoscopy and as histologically differentiated adenocarcinoma on biopsy), who were also examined by EUS. We retrospectively classified EUS features of the third layer (submucosa) into five groups: (1) irregular narrowing, (2) budding sign, (3) multiple echo-free spots, (4) unclear, and (5) no changes. In endoscopically diagnosed gastric mucosal cancer, 16 of the 78 lesions were associated with histologic submucosal invasion. EUS features that were associated with a high incidence of histological submucosal tumor invasion were irregular narrowing (submucosal invasion, 60.0%) and the budding sign (85.7%), and 90.9% of lesions with either of these features had submucosal invasion of tumors when tumorous changes in the third layer exceeded 1 mm in depth. Endosonographic irregular narrowing and a budding sign of more than 1 mm in depth in the third layer are useful for the diagnosis of submucosal invasion in gastric cancers that are diagnosed as mucosal cancers without ulcerous change on endoscopy. PMID- 10832667 TI - Influence of Helicobacter pylori infection on development of stress-induced gastric mucosal injury. AB - Immediately after the Great Hanshin Earthquake in Kobe in 1995, the recurrence rate of peptic ulcer in patients infected with Helicobacter pylori was higher than that in patients in whom H. pylori had been eradicated. We evaluated the influence of H. pylori infection on stress-induced gastric mucosal injury in Mongolian gerbils and C57BL/6 mice. These animals were immersed in water for 30, 120, and 720 min 12 weeks after inoculation with H. pylori, and then killed to assess gastric mucosal damage, and to measure cytokine production (interleukin [IL]-1beta, IL-4, IL-6, and IL-10; interferon [IFN]-gamma; and tumor necrosis factor [TNF]-alpha) in the gastric tissue of the mice. The stress treatment for 30 min resulted in a significantly higher bleeding rate and bleeding index among infected gerbils and mice compared with results in uninfected animals. Conversely, the bleeding and ulcer indexes were significantly higher in uninfected gerbils after 720 min of the stress treatment than in infected gerbils. Prior to the stress treatment, gastric IL-1beta and IFN-gamma production was significantly higher in the infected group than in the uninfected group. After 120 min of the stress treatment, TNF-alpha production was increased in the infected group, and IL-1beta and IL-10 production was increased in the uninfected group. However, the production of these cytokines showed no change at 30 min of the stress treatment. These results suggest that H. pylori infection influences the development of gastric mucosal injury in the early phase of stress exposure; cytokines do not play a major role in this process. PMID- 10832668 TI - Preventive efficacy of butyrate enemas and oral administration of Clostridium butyricum M588 in dextran sodium sulfate-induced colitis in rats. AB - Butyrate enemas have been reported to be effective in ulcerative colitis. However, long-term use is difficult because of the troublesome procedure and the unpleasant smell. We therefore investigated the effects of the oral administration of Clostridium butyricum M588 (CBM588), an enterobacterium producing butyrate, in dextran sodium sulfate (DSS)-induced colitis in rats. First, we confirmed the effects of pre-treatment with a butyrate enema on DSS colitis. We then studied the efficacy of oral administration of CBM588 which was started 1 week prior to the induction of DSS colitis. In the CBM588 group, the ulcer index and myeloperoxidase (MPO) activity in the distal colon were significantly lower than in the control group. Proliferating cell nuclear antigen (PCNA) immuno-positive cells were increased around the ulcer in the CBM588 group. In regard to the contents of the cecum and colon, the proportions of Lactobacillus and Eubacterium were increased in the cecum in the CBM588 group. Further, there were significant increases of n-butyrate, propionate, and acetate concentrations in the cecum in the CBM588 group. These results indicated that the oral administration of CBM588 alleviated DSS-induced colitis, and may be useful instead of butyrate enema. PMID- 10832669 TI - Efficacy of ribavirin in patients with chronic hepatitis B. AB - This study aimed to evaluate the efficacy and safety of a 6-month course of ribavirin (Rb) (1200 mg/day) in the treatment of chronic hepatitis B (CHB). Sixty patients with CHB were randomly assigned in a double-blind placebo (Pl) controlled study; 30 patients received oral Rb (1200 mg/day) and 30 received Pl for 24 weeks. Patients were hepatitis B surface antigen (HBsAg); hepatitis B envelope antigen (HBeAg); and hepatitis B virus (HBV)-DNA-positive, with alanine aminotransferase (ALT) levels 1.5 times higher than normal values. Clinical evaluations and laboratory tests were carried out at regular intervals; tests included total blood cell count, liver function tests, and HBV serum markers. Baseline and control liver biopsies were carried out. HBeAg seroconversion occurred in 50.0% of the patients in the Rb group (vs 6.6% in the Pl group; P = 0.00019); HBV DNA negativization occurred in 33.3% in the Rb group (vs 6.6% in the Pl group; P = 0.009); and improvement in the necroinflammatory index occurred in 53.3% in the Rb group (vs 23.3% in the Pl group; P = 0.02). The drug was well tolerated; the most important side effect in the Rb group was hemoglobin reduction, which was reversible once the treatment was stopped. Ribavirin was an effective treatment, demonstrated by decreased ALT levels, alleviation of histological damage, seroconversion of HBeAg, and HBV-DNA negativization; Rb may be an alternative agent in the treatment of CHB, without significant side effects. PMID- 10832670 TI - Hepatic vascular side effects of styrene maleic acid neocarzinostatin in the treatment of hepatocellular carcinoma. AB - Styrene-maleic acid neocarzinostatin (SMANCS) sometimes causes hepatic vascular side effects, including arterial stricture, obstruction, and arterio-portal shunt. A total of 128 intra-arterial SMANCS injection treatments, performed for 89 patients with hepatocellular carcinoma, were analyzed to determine the relationship between angiographic findings and subsequent hepatic vascular injuries. After SMANCS therapy, hepatic arterial stricture or obstruction occurred in 5 patients (5/128; 3.9%), arterio-portal shunting in 12 (12/128; 9.4%), liver shrinkage in 4 (4/128; 3.1%), and cholangitis or biloma in 2 (2/128; 1.6%). Among 23 patients whose plain abdominal X-ray films just after SMANCS injection showed Lipiodol retention in the hepatic artery, 5 patients developed arterial obstruction, 10 developed arterio-portal shunt, and 2, cholangitis or biloma. Among 26 patients with Lipiodol retention in the portal vein, 4 developed hepatic lobe atrophy with aggravation of liver function. Among 3 patients with Lipiodol retention in both the hepatic artery and the portal vein, 1 developed arterio-portal shunt. In 76 treatments without excessive Lipiodol retention, only 1 of the patients developed arterio-portal shunt. Excessive retention of Lipiodol in hepatic vascular beds just after SMANCS therapy was significantly associated with future vascular side effects (22/52 vs 1/76; P < 0.0001). Lipiodol retention in arteries just after SMANCS injection was closely associated with subsequent arterial obstruction or arterio-portal shunt, and Lipiodol retention in the portal vein was related to subsequent hepatic lobe atrophy. PMID- 10832671 TI - Taurine preserves gap junctional intercellular communication in rat hepatocytes under oxidative stress. AB - Gap junctional intercellular communication (GJIC) between hepatocytes is important for the maintenance of differentiated liver functions. Taurine is known to be cytoprotective, and is used clinically to improve liver functions. We evaluated the effect of taurine on GJIC in hepatocyte doublets under oxidative stress. Hepatocyte doublets were isolated from female Wistar rats, using a collagenase perfusion technique, and cultured in Leibovitz-15 medium containing fetal bovine serum (10%). H2O2 (2 mM) and/or taurine (0.1-1 mM) were added 2 h after inoculation, and the culture was incubated for 3 h. Fluorescent dye (Lucifer Yellow CH) coupling between adjacent cells was evaluated by microinjection. The distribution and quantity of connexin 32 (Cx32) in hepatocytes were detected using indirect immunofluorescence analysis and Western blotting. Steady state mRNA levels of Cx32 were detected by Northern blotting. The percentage of dye coupling 5 h after inoculation was 88 +/- 6.3% in the control. however, this was decreased to almost half the control value by H2O2. Taurine prevented the decrease caused by H2O2 in a dose-dependent manner. Immunofluorescence analysis for Cx32 demonstrated numerous punctate fluorescent spots along the intercellular plasma membrane in controls, which were significantly decreased by H2O2. Taurine prevented the decrease of Cx32. Western blot analysis also showed the decrease of Cx32 protein levels by H2O2 treatment, which decrease was prevented by taurine. Interestingly, H2O2 and/or taurine treatments did not affect Cx32 mRNA levels. Our findings indicated that H2O2 treatment decreased GJIC between hepatocytes, most likely due to augmenting the degradation of Cx32 proteins, whereas taurine prevented this process. This effect of taurine is beneficial for the preservation of differentiated functions in the liver under oxidative stress. PMID- 10832672 TI - Effects of cyclosporin A on water-immersion stress-induced gastric lesion and gastric secretion in rats. AB - Cyclosporin A is an immunosuppressive agent which is well known as a specific inhibitor of calcineurin (protein phosphatase 2B). In this study, we investigated the effects of cyclosporin A on water-immersion stress-induced gastric ulcer formation and gastric acid secretion in rats. We also examined the localization of calcineurin immunohistochemically. Calcineurin was specifically expressed in gastric parietal cells and chief cells of the gastric mucosa. The intraperitoneal administration of cyclosporin A dose-dependently suppressed the development of gastric mucosal lesions induced by water-immersion stress and inhibited gastric acid secretion, as assessed by pylorus ligation. These results indicated that calcineurin may play an important role in gastric acid secretion. PMID- 10832673 TI - Details of hyperplastic polyps of the stomach shrinking after anti-Helicobacter pylori therapy. AB - The precise etiology of hyperplastic polyps of the stomach is unknown, but recent studies suggest that they arise as a consequence of inflammation occurring in intimate association with Helicobacter pylori infection. The process of polyp regression after anti-H. pylori therapy, however, is unclear. Here we report a patient with large hyperplastic polyps of the stomach that regressed markedly after anti-H. pylori therapy. Histological examination of the regressed polyps revealed a decrease in the height of the hyperplastic foveolar epithelium and a decrease in the amount of inflammatory cell infiltration in the stroma. In addition, the percentage of Ki-67-positive hyperplastic epithelial cells markedly decreased after anti-H. pylori therapy, indicating that the epithelial cell proliferation rate had markedly decreased after treatment. At the same time, the degree of cyclooxygenase-2 expression in epithelial cells in the polyps decreased after treatment. Because cyclooxygenase-2 is expressed at sites of inflammation or neoplasm, these findings are consistent with a decrease in inflammatory cell infiltration, and represent resolving inflammation. PMID- 10832674 TI - Heterotopic gastric mucosa in the gallbladder. AB - We report on a case of heterotopic gastric mucosa in the body of the gallbladder. A 39-year-old man, who was asymptomatic, visited our hospital because of a polypoid lesion in the gallbladder, discovered during a routine health screening. Ultrasonography (US) revealed a broad-based polypoid lesion 1.7 cm in diameter in the body of the gallbladder, which was free of gallstones. The gallbladder mass was faintly enhanced by helical computed tomography. Laparoscopic cholecystectomy was performed because of the possibility of malignancy. The specimen revealed a 1.7 x 1.3 cm polypoid lesion with deep delle in the body, with no gallstones in the gallbladder. Intraoperative frozen examination yielded a diagnosis of hyperplastic polyp of the gallbladder. Histologically, the polypoid lesion consisted of gastric fundic glands located in the whole wall of the gallbladder. The surrounding mucosa consisted of almost normal epithelium without any metaplastic changes. Postoperative technetium 99m-pertechnetate scintigraphy demonstrated no evidence of gastric heterotopia elsewhere in the body. We also review 18 other reports of heterotopic gastric mucosa in the gallbladder in the Japanese medical literature. PMID- 10832675 TI - Non-traumatic gas gangrene in the abdomen: report of six autopsy cases. AB - Six autopsy cases of non-traumatic gas gangrene in the abdomen are reported. Five of the six were caused by clostridia, as identified by culture or histology. There were associated underlying diseases, such as alcoholism, liver cirrhosis, diabetes mellitus, and malignant disease. Three of the six patients had gas gangrene in the liver. Bacterial proliferation and gas accumulation were found in the sinusoids of the liver, and congestion and edema with extensive gas embolism were found in the lungs. Pulmonary gas embolism was considered to be the direct cause of death in these three patients. The other three patients had intestinal clostridial gas gangrene, with alcoholism as an underlying condition. None of the six patients was clinically diagnosed as having gas gangrene. We suggest that gas gangrene should be considered in any patient with abdominal infection. A review of 19 autopsy cases of gas gangrene in the abdomen reported in the Japanese literature is also presented. PMID- 10832676 TI - Erythropoietic protoporphyria with severe liver dysfunction and acute pancreatitis. AB - A case of erythropoietic protoporphyria associated with severe hepatic dysfunction and acute pancreatitis is reported. The patient, a 33-year-old man, was admitted to our hospital complaining of upper abdominal pain, nausea, and vomiting of 3 days' duration. Laboratory tests on admission demonstrated liver dysfunction, anemia, and thrombocytopenia. On the third hospital day, the intensity of the upper abdominal pain increased, concomitantly with elevated levels of serum amylase. Ultrasonography and computed tomography scanning revealed a slightly enlarged pancreas. During this episode, he also complained of various neurological symptoms, including reduced mental alertness, weakness of extremities, constipation, profound sweating, and urinary retention. Porphyrin studies demonstrated markedly elevated erythrocyte and fecal protoporphyrin levels. Laparoscopic findings obtained after the attack subsided were compatible with porphyric liver cirrhosis. We therefore concluded that neurologic disorders and acute pancreatitis could develop in patients with erythropoietic protoporphyria with severe liver dysfunction. PMID- 10832677 TI - Lymphoepithelial cyst of the pancreas with sebaceous differentiation. AB - We recently encountered a patient with a lymphoepithelial cyst of the pancreas with sebaceous differentiation. We sought to compare the characteristics of this patient with those previously reported in order to foster a keener understanding of this rare clinical entity. After reviewing the present patient's case in detail, we conducted a comprehensive review of the English-language literature and analyzed the clinical characteristics of reported cases of lymphoepithelial cysts. Our patient was an asymptomatic 60-year-old man who presented with an incidental finding of a cystic lesion in the tail of the pancreas documented by computed tomography. The cyst was enucleated, and was found to contain keratinized material. It was lined by squamous epithelium with small sebaceous glands, and surrounded by lymphoid tissue with germinal centers. Of 33 reported cases, only 6 (18%) contained sebaceous glands. In all patients who underwent operation, the cysts were easily resected, and the outcome was favorable. Lymphoepithelial cyst of the pancreas is rare, and may be difficult to differentiate from cystic neoplasms preoperatively. Therefore resection is indicated. The diagnosis, however, can be confirmed by careful histologic review, and the prognosis is excellent. PMID- 10832678 TI - Endoscopic ultrasonography for determining the depth of cancer invasion in gastric cancer. PMID- 10832679 TI - Is ribavirin treatment really effective for chronic hepatitis B? PMID- 10832680 TI - How can we avert hepatic vascular complications in SMANCS therapy for hepatocellular carcinoma? PMID- 10832681 TI - The resiliency of the corneal endothelium to refractive and intraocular surgery. AB - PURPOSE: To describe stress factors (phenylephrine and contact lenses) from the corneal epithelium that can affect the corneal endothelium, and to describe the effects of refractive and intraocular surgery on the corneal endothelial structure and function. METHODS: Significant clinical and experimental publications are reviewed and recent experiments conducted in the author's laboratory to describe the corneal endothelial stresses. RESULTS: The corneal epithelium serves as a barrier to topical phenylephrine (2.5-10%). In a compromised epithelium, topical phenylephrine will cause drug-induced stromal edema and endothelial vacuolization. Contact lenses are capable of stimulating the epithelial arachidonic acid cascade to release 12(R)hydroxyeicosatetraenoic acid (12(R)HETE) and 8(R)hydroxy-hexadecatrienoic acid (8(R)HHDTrE) to cause endothelial Na+/K+ adenosine triphosphatase (ATPase)-inhibition and polymegethism. Specular microscopy of the corneal endothelial cells after refractive surgery (photorefractive keratectomy [PRK], laser in situ keratomileusis [LASIK], intrastromal rings [INTACs]) has shown that there is minimal effect. However, laser ablation of the stroma within 200 microm of the corneal endothelium will result in endothelial cell structural changes and the formation of the amorphous substance deposited onto Descemet's membrane. Phacoemulsification with a high flow of the irrigation solution can alter the endothelial surface glycoprotein layer. Lidocaine hydrochloride (1%) used as intracameral anesthesia readily diffuses through the corneal endothelium, resulting in stromal uptake and endothelial cell swelling. With phacoemulsification, however, the washout of lidocaine from the cornea (T1/2, 5 minutes) and iris (T1/2, 9 minutes) occurs quickly. Corneal endothelial wound healing after keratoplasty occurs in the following sequence: migration of endothelial cells, development of tight junctions, and the formation of Na+/K+ ATPase pump sites. CONCLUSIONS: Corneal endothelial resiliency is due to the increased peripheral endothelial cell number for migration, the ability of endothelial cells to form tight junctions to maintain the endothelial barrier, the increase in endothelial Na+/K+ ATPase pump sites under stress, and the ability of the corneal endothelial cells to shift their metabolism of glucose to the hexose monophosphate shunt for the production of nicotinamide adenine dinucleotide phosphate (NADPH) and membrane repair. All of these factors are important, along with the aqueous humor sodium concentration, which establishes the osmotic gradient for corneal deturgescence and transparency. PMID- 10832682 TI - Tetracyclines and the treatment of corneal stromal ulceration: a review. AB - PURPOSE: To demonstrate the potential value of tetracyclines in the treatment of corneal ulceration after moderate to severe ocular chemical injuries. METHODS: Review of published materials describing landmarks in the development of tetracyclines as matrix metalloproteinase inhibitors in ophthalmology and related disciplines. RESULTS: Tetracyclines can protect the cornea against proteolytic degradation after moderate to severe ocular chemical injury. They inhibit matrix metalloproteinases by mechanisms independent of their antimicrobial properties, primarily through restriction of the gene expression of neutrophil collagenase and epithelial gelatinase, suppression of alpha1-antitrypsin degradation, and scavenging of reactive oxygen species. CONCLUSION: Oral tetracyclines can be used along with topical tetracycline preparations and other therapeutic agents to inhibit collagenolytic degradation of the cornea after moderate to severe ocular chemical injuries. PMID- 10832683 TI - Surgical management of ocular surface squamous neoplasms: the experience from a cornea center. AB - PURPOSE: To report the surgical outcomes of treatment for ocular surface squamous neoplasms. METHODS: Retrospective review of squamous neoplasms of the ocular surface managed at a cornea center over a 15-year period. Surgical treatment was divided into two methods. The procedure was identical within each group. One group of patients was treated with simple excision of the lesion. The second group of patients was treated with excision followed by focal cryotherapy to the involved limbus and/or conjunctival margin. Follow-up is reported. RESULTS: A total of 28 lesions were reviewed. Included were 20 primary tumors and 8 recurrent tumors referred following initial treatment at other centers. Histopathologic diagnoses included 1 actinic keratosis, 7 dysplasias, 18 carcinomas in situ, and 2 invasive squamous cell carcinomas. The rate of recurrence for primary tumors was 28.5% with simple excision and 7.7% for excision combined with cryotherapy (p = 0.27). The rate of recurrence for recurrent tumors was higher: 16.6% for tumors treated with excision and cryotherapy and 50% (one in two) for lesions treated with simple excision (p = 0.46). CONCLUSION: Simple excision of ocular surface squamous neoplasms appears to result in a higher recurrence rate when compared to excision with adjunctive cryotherapy. Judicious cryotherapy to the involved limbus and conjunctival margins results in recurrence rates comparable with other, more extensive cryotherapy procedures. This technique of cryotherapy has very few adverse effects on the eye. PMID- 10832684 TI - Amniotic membrane transplantation or conjunctival limbal autograft for limbal stem cell deficiency induced by 5-fluorouracil in glaucoma surgeries. AB - PURPOSE: To determine if human amniotic membrane transplantation or limbal stem cell transplantation is effective to restore the corneal surface with partial or total limbal stem cell deficiency, respectively, caused by 5-fluorouracil (5-FU) toxicity after glaucoma surgeries. METHODS: Partial and total limbal stem cell deficiency was confirmed by impression cytology as the cause of reduced vision and corneal surface breakdown in a 69-year-old man and a 67-year-old man, respectively, who both had received a total of 105 mg 5-FU injections. Amniotic membrane transplantation or conjunctival limbal autograft was performed for corneal surface reconstruction, respectively. RESULTS: For a period of 15 months of follow-up, the visual acuity improved, and their corneal surfaces remained avascular, smooth, and without recurrence of limbal stem cell deficiency. CONCLUSION: Limbal stem cell deficiency can occur as a late complication for patients receiving 5-FU after glaucoma filtering surgeries. Partial limbal stem cell deficiency can be treated with amniotic membrane transplantation alone, whereas limbal transplantation must be considered as an alternative for total limbal stem cell deficiency to restore the corneal surface integrity and vision. PMID- 10832685 TI - Use of nonpreserved human amniotic membrane for the reconstruction of the ocular surface. AB - PURPOSE: To describe the use of nonpreserved human amniotic membrane (NP-AMT) as an alternative to preserved human amniotic membrane (AMT) for the reconstruction of the ocular surface in several diseases. METHODS: NP-AMT was used in the treatment of five patients with the following diseases: noninvasive conjunctival squamous cell carcinoma, corneal persistent epithelial defect, severe alkali burn, near total limbal deficiency secondary to multiple surgeries, and ocular cicatricial pemphigoid. In some cases, a limbal autograft or allograft was employed simultaneously, sutured on top of the NP-AMT. All sutures were made with 10-0 Nylon and were removed at two weeks. RESULTS: Ocular surface was satisfactorily reconstructed, eyes were quiet, and patients were comfortable despite prolonged deepithelialization in some cases. There was a case of a limbal autograft ischemia--in the burned patient--that caused partial corneal conjunctivalization. Initially, the NP-AMT looks thickened but thins around the fifth day and looks similar to AMT. CONCLUSION: Results using NP-AMT are similar to those of AMT. It is a good alternative and it is easily obtained in places were AMT is not available or is too expensive to procure. PMID- 10832686 TI - Evaluating pterygium severity: a survey of corneal specialists. AB - PURPOSE: Accurate and reliable evaluation techniques are essential for clinical and epidemiologic studies. This survey of corneal specialists was designed to lay a foundation for the further development of methods for evaluating and staging pterygium. METHODS: In a self-administered, mailed questionnaire, 213 corneal specialists rated the importance of nine symptoms, nine signs, and nine clinical tests for the severity of primary pterygium. Severity was defined as the present need for surgical intervention. RESULTS: The most important factors for determining primary pterygium severity were the extent of encroachment onto the cornea, decreased visual acuity, restricted ocular motility, and increased rate of growth. Many patient symptoms were rated as moderately to highly important. The questionnaire was shown to have good response reliability by test-retest comparisons. Cronbach's alpha was 0.89, which indicates very good internal consistency reliability. CONCLUSION: The survey identifies the priorities of experts in determining the severity of pterygium. More precise and clearly defined evaluation methods will enhance future clinical and epidemiologic studies of pterygium. The ranked list of pterygium signs, symptoms, and tests can serve as a guide for developing pterygium evaluation methods in the future. There is a need for a method that accurately and precisely quantifies the distance of pterygium encroachment onto the cornea and the pterygium progression rate. Furthermore, there is a need for an assessment of patient symptoms. PMID- 10832687 TI - A devastating ocular pathogen: beta-streptococcus Group G. AB - PURPOSE: To report the clinical findings, treatment, and outcomes of four cases of beta-streptococcus Group G (BHS-G) ocular infection. METHODS: The medical and microbiologic records of four cases of BHS-G ocular infection were retrospectively reviewed. RESULTS: Two cases of BHS-G endophthalmitis and two cases of BHS-G keratitis were recorded. Three patients developed fulminant infection within 12 hours of the onset of symptoms. One patient's history was incomplete. One patient developed endophthalmitis from a contaminated donor button; another following cataract surgery. One developed keratitis in a keratoplasty suture tract; and another patient developed a corneal abscess after being struck with a tree branch. The patient with the contaminated donor button developed overwhelming endophthalmitis resulting in no light perception vision, severe pain, and evisceration. The postoperative cataract patient developed a purulent endophthalmitis and is still hypotonus with light perception vision. The second keratitis patient developed a significant suture abscess with marked stromal loss but eventually healed. The traumatic keratitis patient developed a large ulcer with hypopyon and descemetocele but was lost to follow-up. CONCLUSIONS: This is the first report of a series of BHS-G ocular infections. The ocular infections were characterized by rapid onset, extreme inflammation, and- despite in vitro antibiotic sensitivity--a poor or sluggish response to antibiotic therapy. PMID- 10832688 TI - Clinical and microbiological profile of Bacillus keratitis. AB - PURPOSE: To examine the clinical and microbiological profile of Bacillus keratitis. METHODS: A retrospective review was done of all medical and laboratory records of patients with infectious keratitis in an urban tertiary level eye-care center in South India between January 1991 and June 1997. RESULTS: Nineteen eyes of 17 patients having microbiologically proven Bacillus keratitis were reviewed. The mean age of the patients was 32.64 years (range, 3-70). The duration of symptoms ranged from 1 day to 3 months, with 11 eyes seen within a week of onset of symptoms. Trauma (five eyes), lagophthalmos (two eyes), topical corticosteroid therapy (one eye), bullous keratopathy (two eyes), previous corneal scars (two eyes), and diabetes (one eye) were identified as predisposing factors. Severe corneal features, disproportionate to the duration of symptoms, were present in most of the eyes. Gram stain of corneal scrapings showed variably stained bacilli in eight (42.1%) cases. Polymicrobial infection was present in six eyes (two fungal, four bacterial). Of the 16 isolates tested for in vitro antibiotic susceptibility, 100% were sensitive to gentamicin, 15 (93.75%) were sensitive to ciprofloxacin and norfloxacin, 14 (87.5%) were sensitive to chloramphenicol, and 10 (62.5%) were sensitive to cefazolin. Whereas 12 (63.1%) eyes required only medical therapy, adjunctive procedures were required in seven (36.8%) eyes. The ulcers healed (mean time to healing, 37.4+/-28.6 days) in 16 eyes (lost to follow up, three). Visual acuity had improved after treatment in 10 (71.4%) of 14 eyes in whom vision could be recorded. CONCLUSION: Bacillus is an unusual pathogen in the clinical setting of infectious keratitis. The infection is mostly amenable to treatment with commonly used antibiotics, and the final outcome is often satisfactory. PMID- 10832689 TI - Spectrum of fungal keratitis at Wills Eye Hospital, Philadelphia, Pennsylvania. AB - PURPOSE: To report the spectrum of fungal keratitis at Wills Eye Hospital, Philadelphia. METHODS: We reviewed the records of 24 cases of culture-positive fungal keratitis treated from January 1991 to March 1999 at Wills Eye Hospital. Risk factors, fungal identification, antifungal treatment, and outcomes were evaluated. RESULTS: The study included 24 eyes (24 patients). Fourteen patients (58.3%) were female. The mean age was 59 years (range, 19-86 years). Predisposing factors included chronic ocular surface disease (41.7%), contact lens wear (29.2%), atopic disease (16.7%), topical steroid use (16.7%), and ocular trauma (8.3%). Early identification of fungal elements was achieved by staining of corneal scrapings in 18 cases (75%). Half of the cases (12 eyes) had corneal infections caused by yeast, and the other half by filamentous fungi. Candida albicans was the most commonly isolated organism (45.8%), followed by Fusarium sp (25%). Natamycin and amphotericin B were the topical antifungals most frequently used, while systemic treatment commonly used included fluconazole, ketoconazole, or itraconazole. Six patients (25%) had penetrating keratoplasty during the acute stage of infection. After a mean follow-up of nine months, 13 eyes (54.1%) had the best corrected visual acuity 20/100 or better. CONCLUSIONS: In contrast to other studies from the northern United States, we found Fusarium sp the most commonly isolated filamentous fungus. In our series, C. albicans was the most frequent cause of fungal keratitis, and a past history of ocular trauma was uncommon. PMID- 10832690 TI - Nontraumatic corneal perforation. AB - PURPOSE: To study the predisposing conditions, treatments, and visual outcomes of nontraumatic corneal perforations. METHODS: A retrospective chart review was conducted of all nontraumatic corneal perforations seen between January 1992 and December 1998, with > or = 3 months of follow-up, at the Cornea Service Wills Eye Hospital. RESULTS: A total of 40 nontraumatic corneal perforations was analyzed. Sixty-two percent of the cases were female. At presentation, 35 of 40 eyes (87.5%) had best corrected visual acuity of 20/200 or worse. The most common diseases associated with perforations were keratoconjunctivitis sicca (12 eyes, 30%), bacterial keratitis (6 eyes, 15%), exposure keratopathy (5 eyes, 12.5%), and herpes simplex virus (HSV) keratitis (4 eyes, 10%). Visual acuity improved > or = 2 Snellen lines in 3 of 8 eyes (37.5%) treated with penetrating keratoplasty, 5 of 14 eyes (35.7%) treated with tissue adhesive, and 1 of 12 eyes (8.3%) given medical treatment. After allowing for the different levels of presenting vision, treatment modality was not significantly related to final visual outcome. CONCLUSION: Keratoconjunctivitis sicca is the most common underlying disease associated with nontraumatic corneal perforation. Corneal perforations were managed successfully using tissue adhesive, medical therapy, or penetrating keratoplasty. Treatment depended on the characteristics of the perforation and on the visual potential of the eye. PMID- 10832691 TI - Reverse geometry contact lens wear after photorefractive keratectomy, radial keratotomy, or penetrating keratoplasty. AB - PURPOSE: To determine if a super diffusion coefficient of lens/lens thickness (Dk/L) reverse geometry gas permeable (steep peripheral and flatter central curve) contact lenses can be successfully worn after excimer photorefractive keratectomy (PRK), radial keratotomy (RK), or penetrating keratoplasty (PK). METHODS: Patients with residual ametropia after PRK, RK, or PK were fitted with reverse geometry lenses (Plateau lens in SF-P material; Menicon USA, Inc, Clovis, CA, U.S.A.). Contact lens fit characteristics and comfort were assessed. Lens centration, visual quality, and ocular surface status were graded and visual acuity charted. RESULTS: Thirteen eyes of 11 patients were fitted; eight eyes with PRK, one eye with RK, and four eyes with PK previously performed. The mean follow up of the patients was six months. The visual acuity prior to lenses ranged from 6/12 to counting fingers, and the acuity with lenses ranged from 6/6 to 6/30. Eight of the 11 patients wore the lenses the whole day without problems; 3 patients discontinued lens wear due to discomfort or unsatisfactory vision. CONCLUSIONS: The combination of super Dk/L and reverse geometry lenses facilitate lens wear and is associated with good visual acuity. PMID- 10832692 TI - Centration of donor trephination in human corneal transplantation. AB - PURPOSE: To develop and evaluate a new method to quantify centration of the trephined donor cornea relative to the limbus. METHODS: After human donor corneas were trephined for penetrating keratoplasty, the remaining corneoscleral discs were stained and subjected to image analysis. The centration of the excised donor cornea relative to the limbus was calculated by measuring their centroids from the "captured" images. RESULTS: Fifty-two corneoscleral discs were analyzed. The average deviation from the centre was 0.32 mm (SD, 0.18 mm). Neither surgeon nor the type of trephine significantly influenced the mean centroid deviation. CONCLUSION: We have developed and evaluated a method to quantify centration of human donor cornea. In a small series, decentration did not correlate significantly with either the surgeon or the trephine. PMID- 10832693 TI - Penetrating keratoplasty for keratoconus. AB - PURPOSE: We performed a retrospective study of patients with keratoconus who underwent penetrating keratoplasty at the University of California, Davis, during the years 1983-1996 to analyze subsequent visual acuity and the need for optical correction. METHODS: We reviewed 123 eyes of 94 patients and collected data including best corrected visual acuity (BCVA) and uncorrected visual acuity (UCVA), type of correction (contact lens vs. spectacles), incidence of rejection, and other complications. Data were obtained at 12 and 18 months postoperatively. RESULTS: There was a significant improvement of the BCVA between 12 and 18 months (p < 0.05) and no significant improvement in UCVA between at the same times (p = 0.222). At 12 months postoperatively, 84%, and at 18 months, 87% of patients achieved 20/40 or better BCVA. At 18 months, 47% of eyes were fit with contact lenses, and 30%, with spectacles. Mean spherical refraction was -4.13 D +/- 4.41 standard deviation (SD) at 12 months and -4.09 D +/- 3.86 SD at 18 months, whereas mean cylinder was 2.52 D +/- 2.45 SD and 2.67 D +/- 2.04 SD, respectively. Of the eyes, 17.9% had at least one graft rejection, although rejection episodes did not significantly influence the incidence of 20/40 vision (p = 0.084). Combined nonrejection complications did not significantly influence incidence of 20/40 or better vision at 18 months (p > 0.10). CONCLUSION: This study reaffirms that the results for keratoplasty in keratoconus are very positive and emphasizes that ophthalmologists should counsel patients about the likelihood of the need for spectacle or contact lens correction. Our data demonstrate that the majority of patients require optical correction for functional visual acuity after keratoplasty. PMID- 10832694 TI - Corneal triple procedure: indications, complications, and outcomes: a developing country scenario. AB - PURPOSE: We report the indications, complications, and outcomes of 104 corneal triple procedures in our institute. METHODS: Patient records of 104 consecutive cases of corneal triple procedure (penetrating keratoplasty with extracapsular cataract extraction and intraocular lens implantation) performed by experienced corneal surgeons between January 1992 and December 1997 were retrospectively reviewed. Relevant preoperative, operative, and postoperative data were collected. The outcome was assessed by the graft clarity and visual acuity at the last visit. Survival analysis of these grafts was determined by Kaplan-Meier method. RESULTS: Of 104 patients, 70 were men and 34 were women. Mean age of these patients was 48.5+/-17.1 years (range, 1-75 years). Corneal scarring with cataract was the reason for surgery in 69 (66.4%) cases. The intraoperative complications included vitreous upthrust in seven (6.9%) cases and posterior capsular dehiscence in three (2.9%) cases. The most common early postoperative complications were increased intraocular pressure in 19 (18.3%) and increased anterior chamber reaction in 25 (24%) cases. Posterior capsular opacification was seen in 26 (25%), graft reaction in 15 (14.4%), and secondary glaucoma in 15 (14.4%) cases. These formed the important late postoperative complications. The average postoperative follow-up was 23.7+/-17.6 months (range, 1.6-79.4 months); at which time 72% of the grafts remained clear. At last follow-up, 40% of patients had a visual acuity of > or = 20/40. CONCLUSIONS: Corneal scarring with cataract is the most common reason for triple procedure in this part of the world. This is a safe surgical procedure with good graft clarity and reasonable visual recovery. PMID- 10832695 TI - The lids influence on corneal shape. AB - BACKGROUND: The lids are in contact to the corneal surface in normal viewing. Refractive surgery, however, is performed with a speculum in place and thus the lids are not in contact with the cornea. PURPOSE: To investigate central corneal shape with and without the lids touching the corneal surface. METHODS: The technique consists of exporting the point cloud of the topography scanner into computer software that recreates the corneal surface. The technique of surface modeling was used to form central corneal elevation height difference maps of four normal eyes with and without lid contact to the corneal surface. RESULTS: Surface modeling demonstrated the trend that the lids influence the corneal shape. CONCLUSION: The lids distort corneal shape. PMID- 10832696 TI - Phototherapeutic keratectomy for Schnyder's crystalline corneal dystrophy. AB - PURPOSE: To describe the treatment of Schnyder's crystalline dystrophy (SCD) with excimer laser phototherapeutic keratectomy (PTK). METHODS: Chart review of three patients (four eyes) with SCD treated with PTK between March 1992 and December 1998. Pre- and posttreatment visual acuity, subjective glare, photophobia, manifest refraction, ultrasound pachymetry, hyperopic shift, and biomicroscopic findings were studied. Brightness acuity testing (BAT) was evaluated in two eyes. RESULTS: Four eyes were treated during the study period. The average best corrected visual acuity (BCVA) improved from 20/175 to 20/40 under bright conditions. All patients reported subjective improvement in glare and photophobia. No visually significant recurrence was observed during the follow-up period which ranged from seven months to three years. The mean spherical equivalent refractive shift was +3.28 diopter (D). One patient developed irregular astigmatism from an eccentric ablation. No vision threatening complications were observed. CONCLUSION: PTK can be effective at improving visual symptoms in patients with SCD. It can be a useful therapeutic alternative to lamellar or penetrating keratoplasty in these patients. PMID- 10832697 TI - Identification of antiangiogenic and antiinflammatory proteins in human amniotic membrane. AB - PURPOSE: To identify the potential antiangiogenic and antiinflammatory proteins expressed in human amniotic membrane tissue. METHODS: Human amniotic epithelial and mesenchymal cells were isolated from human amniotic membranes by sequential trypsin and collagenase digestion. Total RNAs were harvested from freshly obtained human amniotic epithelial and mesenchymal cells. Antiangiogenic and antiinflammatory proteins were detected by the reverse transcriptase-polymerase chain reaction (RT-PCR) technique and further confirmed by DNA sequencing of PCR amplified transcripts. The distribution of tissue inhibitors of metalloproteinase (TIMPs) were studied further by immunohistochemistry performed on paraffin embedded amniotic membrane tissue. RESULTS: RT-PCR results showed that both human amniotic epithelial and mesenchymal cells express interleukin-1 receptor antagonist, all four TIMPs, collagen XVIII, and interleukin-10. Thrombospondin-1 was expressed in all of the epithelial cell specimens and in one out of five mesenchymal cell specimens. Furthermore, immunohistochemistry studies performed on freshly prepared amniotic membrane confirmed that all members of the TIMP family were present in epithelial and mesenchymal cells as well as in the compact layer of the amniotic stroma. In cryopreserved amniotic membranes, positive staining was seen in residual amniotic cells and stroma. CONCLUSIONS: Human amniotic membrane epithelial and mesenchymal cells express various antiangiogenic and antiinflammatory proteins. Some of those proteins also were found in amniotic membrane stroma. These findings may explain in part the antiangiogenic and antiinflammatory effects of amniotic membrane transplantation. PMID- 10832698 TI - Antibacterial activity of anesthetic solutions and preservatives: an in vitro comparative study. AB - PURPOSE: To investigate the antibacterial activity of topical anesthetic solutions and their preservatives individually in vitro, to determine involvement with bacterial growth inhibition. MATERIAL AND METHODS: Proparacaine and tetracaine (in concentrations of 0.125%, 0.25%, and 0.50%), edetate disodium (EDTA), benzalkonium chloride, EDTA + benzalkonium chloride, and sterile saline solution were used. Five microliters of each solution were applied to standard filter paper disks and placed in Mueller-Hinton agar previously inoculated with known strains of Pseudomonas aeruginosa and Staphylococcus aureus. Zones of growth inhibition were measured 24 hours later and analyzed. RESULTS: There were no zones of inhibition in the agar inoculated with P. aeruginosa to all tested solutions. Benzalkonium chloride alone and associated with EDTA inhibited growth of S. aureus. All other solutions did not inhibit S. aureus. CONCLUSION: Preservative-free anesthetic solutions seemed not to interfere with bacterial development in culture media. Benzalkonium chloride alone and associated with EDTA inhibited development of gram positive bacteria, S. aureus, but did not inhibit P. aeruginosa. PMID- 10832699 TI - Thermal and biomechanical parameters of porcine cornea. AB - PURPOSE: New methods in refractive surgery require a considerable understanding of the material "cornea" and are often studied by theoretical modeling in order to gain insight into the procedure and an optimized approach to the technique. The quality of these models is highly dependent on the preciseness of its input parameters. Porcine cornea often is used as a model in preclinical studies because of its similarity to man and its availability. METHODS: The important physical parameters for biomechanical deformation, heat conduction, and collagen denaturation kinetics have been determined for porcine cornea. Experimental methods include densitometry, calorimetry, turbidimetry, tensile tests, stress relaxation, and hydrothermal isometric tension measurements. RESULTS: The density of porcine cornea was measured as p = 1062+/-5 kg/m3, the heat capacity gave c = 3.74+/-0.05 J/gK. The stress-strain relation for corneal strips is represented by a third order approximation where the secant modulus yields about Esec approximately equal to 0.4 MPa for small strains less than 2%. The normalized stress relaxation is described by an exponential fit over time. The denaturation process of cornea is characterized by specific temperatures which can be related to the change of the mechanical properties. Denaturation kinetics are described according to the model of Arrhenius yielding the activation energy deltaEa = 106 kJ/mol and the phase transition entropy deltaS = 39 J/(mol x K). CONCLUSIONS: The established set of parameters characterizes the porcine cornea in a reliable way that creates a basis for corneal models. It furthermore gives direct hints of how to treat cornea in certain refractive techniques. PMID- 10832700 TI - Inhibitory effects of tranilast on the proliferation and functions of human pterygium-derived fibroblasts. AB - PURPOSE: We studied the possibility that tranilast, an antiallergic and antiproliferative drug, may be beneficial for the treatment of pterygium. METHODS: Pterygium-derived cells were identified by immunohistochemical methods. Growth rate of pterygium-derived cells was determined by using a hemocytometer. Chemotaxis was determined in a microchemotaxis chamber. Pterygium-derived cells were cultured on floating collagen gel, and the contracted diameter was measured. Collagen synthesis by pterygium-derived cells was determined by the collagenase digestive method. Tranilast was added to the culture medium at final concentrations of 0, 12.5, 25, 50, and 100 microg/ml. RESULTS: Pterygium-derived cells were stained with anti-prolylhydroxylase and anti-alpha-smooth muscle actin, and identified as fibroblasts. Tranilast inhibited the proliferation and chemotaxis of pterygium-derived fibroblasts, and the collagen-gel contraction induced by these cells, but it exerted no inhibitory action on collagen synthesis by pterygium-derived fibroblasts. CONCLUSION: Tranilast may be useful for suppressing the recurrence and, possibly, the development of pterygium. PMID- 10832701 TI - Adeno-associated and herpes simplex viruses as vectors for gene transfer to the corneal endothelium. AB - PURPOSE: We examined the efficacy and cytopathogenicity of adeno-associated (AAV) and herpes simplex viruses (HSV) as vectors for gene transfer to corneal endothelial cells (CECs). METHODS: Recombinant AAV and HSV were examined for their ability to deliver a lacZ histochemical marker gene to whole-thickness rabbit and human corneas ex vivo. Transgene expression was detected with histochemistry and quantified by a colorimetric assay. RESULTS: Rabbit and human corneas transduced with AAV showed increasing numbers of cells expressing marker gene over a 3- to 4-week period. Using 2.5 x 10(6) or 1.5 x 10(7) infective units for rabbit and human corneal specimens, respectively, approximately 2% of CECs expressed the reporter gene. HSV (10(6) plaque-forming units/specimen) transduced approximately 5% of rabbit and human CECs but showed cytotoxicity. In contrast to the duration of recombinant AAV-mediated lacZ expression, recombinant HSV expression was maximal at day 1 and declined to low levels at day 7. CONCLUSION: AAV is a promising vector, but its usefulness for corneal transduction is currently limited by the technical difficulties preparing high titres. The HSV vector examined is efficient but needs further genetic modification to prolong transgene expression and reduce its toxicity. PMID- 10832702 TI - ATPase-positive dendritic cells in the limbal and corneal epithelium of guinea pigs after extended wear of hydrogel lenses. AB - PURPOSE: It has been said that Langerhans cells can be induced to migrate into the central cornea by various stimuli. We investigated whether extended wear of hydrogel lenses induced the migration of Langerhans cells into the central cornea. METHODS: Guinea-pig eyes were fitted with hydrogel contact lenses, and the number of adenosine triphosphatase (ATPase)-positive dendritic cells at the limbus, peripheral, and central corneal epithelium was quantified after 1, 2, 4, and 8 nights of extended wear. The cells were identified with histochemical procedures using ATPase stain. RESULTS: At baseline, ATPase-positive dendritic cells were seen at the limbus and peripheral cornea, with the density decreasing from limbus to peripheral cornea. There were no cells in the central cornea. With extended wear, cells increased in number at the peripheral cornea from 2 nights onward, and cells were seen in the central cornea from 4 nights onward. CONCLUSION: Extended wear of hydrogel lenses induced migration of ATPase-positive dendritic cells (Langerhans cells) into the central cornea. It is possible that they may play a role in the pathophysiology of some of the adverse events seen with contact lens wear. PMID- 10832703 TI - Renewal of the rabbit corneal epithelium as investigated by autoradiography after intravitreal injection of 3H-thymidine. AB - PURPOSE: To evaluate the renewal time of the rabbit corneal epithelium and to verify whether there are differences in the rate of renewal related to the topography of the cornea. METHODS: After the intravitreal injection of 20 microBq/eye of 3H-thymidine (3H-TdR), the rabbits were killed at variable survival-time intervals (3 hours to 90 days), and their corneas were processed for autoradiography on paraffin and 0.75-microm-thick Epon sections. The frequency of labeled nuclei, per each linear millimeter of epithelium, was estimated on paraffin sections of the whole length of the cornea of rabbits killed 6 hours after injection. The activity of unbound 3H-TdR was measured in the vitreous and aqueous humor. RESULTS: The incorporation of 3H-TdR into newly synthesized DNA ceased between 1 and 2 days after the intravitreal injection. At the shortest time intervals (3 hours to 1 day), the overwhelming majority of the labeled nuclei were located in the basal stratum of the stratified squamous epithelium. Cells of the wing stratum were labeled from 3 days onward. At 14 days, labeled nuclei were visualized only in the inner portion of the surface stratum, whereas their outermost layers were labeled only at 21 days and later. Lightly labeled nuclei could still be detected in the epithelium < or = 90 days after injection. The counts of labeled nuclei did not show significant differences in their frequencies between the periphery and the center of the cornea. The existence of vascularization at the periphery of the corneal stroma and the lack of a conspicuous Bowman's layer made it difficult to characterize morphologically the rabbit limbus with the light microscope. CONCLUSION: It takes >14 days for a daughter cell to migrate from the basal stratum to the outermost layer of the epithelium. Labeled daughter cells were detected < or = 90 days in several layers of the corneal epithelium. Therefore it takes >2 weeks for the complete renewal of the epithelium. Our data suggest that the proliferative capability of the centrally located basal cells is enough to guarantee the renewal of the corneal epithelium under physiologic conditions. PMID- 10832704 TI - Synthetic complementary peptides inhibit a neutrophil chemoattractant found in the alkali-injured cornea. AB - PURPOSE: We have previously presented evidence that the neutrophil chemoattractant, N-acetyl-proline-glycine-proline (N-acetyl-PGP), triggers the initial polymorphonuclear leukocyte (PMN) invasion into the alkali-injured eye. In this study, sense-antisense methodology was used to develop novel complementary peptides to be potential inhibitors of N-acetyl-PGP. METHODS: The polarization assay was used to measure the potential chemotactic response of PMNs to synthetic N-acetyl-PGP, the ultrafiltered tripeptide chemoattractants obtained from alkali-degraded rabbit corneas, or leukotriene B4 (LTB4). Inhibition was expressed as the peptide concentration producing 50% inhibition (ID50) of polarization. Five complementary peptides were tested as potential inhibitors of N-acetyl-PGP: arginine-threonine-arginine (RTR), RTR-glycine-glycine (RTRGG), RTR dimer, RTR tetramer, and alanine-serine-alanine (ASA) tetramer. In addition, the RTR tetramer and both monomeric peptides (RTR and RTRGG) were separately tested for inhibition of the ultrafiltered tripeptide chemoattractants or LTB4. RESULTS: The complementary RTR tetrameric peptide was a powerful antagonist of N-acetyl PGP-induced PMN polarization (ID50 of 200 nM). The RTR dimer was much less potent (ID50 of 105 microM). Both monomeric peptides, RTR and RTRGG, were only antagonistic at millimolar concentrations. The ASA tetramer showed no capacity to inhibit N-acetyl-PGP. The RTR tetramer also inhibited PMN activation by the ultrafiltered tripeptide chemoattractants (ID50 of 30 microM) but had no effect on LTB4. CONCLUSIONS: A complementary peptide (RTR) was designed which is an effective inhibitor of the neutrophil chemoattractant, N-acetyl-PGP. The potency of the RTR complementary peptide is dramatically enhanced by tetramerization. Inhibition of N-acetyl-PGP by complementary peptides offers great promise for control of the inflammatory response in the alkali-injured eye. PMID- 10832705 TI - Collagen pleomorphism in Descemet's membrane of streptozotocin-induced diabetic rats: an electron microscopy study. AB - BACKGROUND: Diabetes mellitus causes ultrastructural changes in various basement membranes. These changes include increase in collagen biosynthesis rate and production of altered collagen. PURPOSE: To evaluate the ultrastructural changes in the corneas of streptozotocin- induced diabetic rats, focusing on Descemet's membrane. METHODS: Sprague-Dawley rats were sacrificed at different intervals after induction of diabetes mellitus by streptozotocin. Ten animals were sacrificed 1, 2, 3, 4, 6, 8, and 12 months after the injection of streptozotocin. Five untreated rats of the same age were used as normal controls and were sacrificed at the same intervals. Ultrathin sections were obtained from the corneas and were examined by transmission electron microscopy. RESULTS: Unusual 120-nm-spaced collagen fibril bundles were found in Descemet's membrane of the diabetic rats as early as the second month following the induction of diabetes. Their concentration and size increased gradually over the follow-up period of 12 months. CONCLUSIONS: Changes in the morphologic features of the collagen within Descemet's membrane may occur in diabetic-induced rats. The appearance of wide spaced collagen fibrils in Decemet's membrane may represent alteration in collagen biosynthesis by the endothelial cells in diabetes or altered assembly of collagen due to increased glycosylation of normal formed collagen. PMID- 10832706 TI - Tarantula keratouveitis. AB - PURPOSE: To report a case of chronic bilateral keratouveitis, which was initiated after contact with a pet tarantula. METHODS: A 16-year-old male presented with a photophobia and redness of his eyes two days after handling a tarantula. He was found to have a number of linear corneal foreign bodies with subepithelial infiltrates. The infiltrates were found at varying levels of the corneal stroma with deposits on the endothelium. The anterior chamber had a mild cellular reaction. RESULTS: He was treated with topical steroid drops, which made him asymptomatic. However, he continued to have a mild persistent keratitis and iritis four months after the onset. CONCLUSION: Tarantula hairs may be associated with a chronic keratouveitis, which is usually self-limiting and responds well to treatment with topical steroids. Tarantula pet owners should be forewarned of the ocular dangers associated with handling these spiders. PMID- 10832707 TI - Recurrent keratoconus in a patient with Leber congenital amaurosis. AB - PURPOSE: Clinical history of a 17-year-old patient with Leber congenital amaurosis (LCA) with histologically proven recurrent keratoconus (KC) two years after corneal transplantation in one eye and a recurrence-like appearance with a more global contour on the other eye four years after corneal grafting is reported. The possible mechanisms for this recurrence are discussed in light of the fact that this is, to the best of our knowledge, the first penetrating keratoplasty reported in LCA. METHODS: Computerized videokeratography (CVKG) and specular microscopy were performed preoperatively. The patient underwent regrafting, and the excised corneal button was examined by light microscopy and transmission electron microscopy. RESULTS: Analysis of CVKG showed a keratoconus like pattern on the right eye, with the left eye demonstrating the aspects usually seen in keratoglobus. Histologic examination revealed the features usually observed in progressed keratoconus. CONCLUSION: Recurrence of keratoconus in a graft has not yet been described after such a short time until now. A "true" recurrence of the disease is postulated; it could be caused by an "aggressive" genetic factor that also leads to the frequent KC in patients with LCA. This mechanism also could explain the high incidence and rapid progress of KC in this disease. PMID- 10832708 TI - Atypical extensive recurrent pterygium associated with type 1 neurofibromatosis: histology and management by corneoscleral-conjunctival allograft. PMID- 10832709 TI - Marginal infiltrative ulcerative keratitis secondary to Churg-Strauss syndrome: a case report. AB - PURPOSE: To describe a patient with keratitis associated with Churg-Strauss syndrome. METHODS: A 44-year-old lady with known Churg-Strauss syndrome was examined with marginal infiltrative ulcerative keratitis. RESULTS: Autoimmune disorders with systemic vasculitis are a major cause of marginal keratitis, and in this patient, the diagnosis of Churg-Strauss was confirmed clinically and pathologically. CONCLUSIONS: Churg-Strauss syndrome should be considered in the differential diagnosis of marginal keratitis. PMID- 10832710 TI - Medical management of Beauveria bassiana keratitis. AB - PURPOSE: To describe a case of Beauveria bassiana keratitis and to discuss the management of this rare condition. METHODS: An 82-year-old woman underwent surgical repair of a graft wound dehiscence. Seven months later, shortly after the removal of sutures, the patient developed a fungal keratitis. B. bassiana was identified as the infecting organism. The patient was treated with topical natamycin and oral fluconazole. RESULTS: Following antifungal therapy, the corneal ulcer was eradicated, but the patient underwent repeat penetrating keratoplasty for decreased vision due to corneal edema. The graft remains clear and visual acuity is substantially improved. CONCLUSION: The medical management of B. bassiana keratitis has previously been unsuccessful. The use of topical natamycin combined with oral fluconazole in the management of this case is discussed. PMID- 10832711 TI - Noncontact tonometry in perforating corneal injuries. AB - PURPOSE: To report on one of the possible complications induced by puff noncontact tonometry and to discuss various aspects such as safety, other optional methods, rationality, and the need for tonometry in cases of perforating corneal injuries. PATIENTS AND METHODS: We report a case of traumatic, self sealed, midperipheral corneal perforation, where puff tonometry was performed upon admission, 30 minutes after the injury. RESULTS: The integrity of the wound was temporarily distorted by the air-jet of the puff tonometer, the wound was opened, and an air-bubble filled the anterior chamber. CONCLUSION: Puff tonometry in patients with self-sealed midperipheral corneal perforation and a negative Seidel test does not seem sufficiently safe during the immediate posttraumatic period. PMID- 10832712 TI - Self-induced corneal crystals: a case report. PMID- 10832713 TI - Is donor age an important determinant of graft survival? PMID- 10832714 TI - New cancer therapy by immunomanipulation: development of immunotherapy for human melanoma as a model system. AB - PURPOSE AND METHODS: T cells play an important role in in vivo rejection of human melanoma. Human melanoma antigens recognized by autologous T cells were identified. These antigens are classified as tissue (melanocyte)-specific proteins, cancer-testis antigens (proteins expressed in normal testis and various cancers), tumor-specific peptides derived from mutations in tumor cells, and others. RESULTS: A variety of mechanisms generating T cell epitopes on tumor cells were discovered. Various clinical observations, including tumor regression observed in adoptive transfer of gp100-reactive T cells suggest that these identified melanoma peptides may function as tumor rejection antigens. Immunodominant common epitopes that could expand melanoma-reactive cytotoxic T lymphocytes (CTLs) in vitro were found in the MART-1 and gp100 antigens. New immunization protocols--including immunization with peptides, recombinant viruses, plasmid DNAs, and dendritic cells pulsed with peptides as well as adoptive transfer of in vitro-generated CTLs by stimulation with antigenic peptides--were developed (phase I clinical trials have been performed in the Surgery Branch of the National Cancer Institute, Bethesda, MD, U.S.A.). Immunization with the gp100(209(210M)) peptide that was modified to have high HLA A2 binding affinity, along with incomplete Freund's adjuvant and interleukin (IL) 2, resulted in a 42% response rate in patients with melanoma. CONCLUSION: These immunotherapies need further improvement due to the mechanisms of tumor escape from T cell responses. PMID- 10832715 TI - Role of apoptosis in wound healing in the cornea. AB - Anterior keratocytes undergo apoptosis in response to epithelial injury. This likely represents mechanical activation of systems that vigilantly monitor the corneal epithelium for injury produced by pathogenic viruses. Keratocyte apoptosis is the first change noted after procedures in which the epithelium is injured and may be an initiator of the subsequent wound healing cascade. The working hypothesis is that inhibition of keratocyte apoptosis will limit the subsequent wound healing response. The keratocyte apoptosis response varies with the specific type of epithelial injury induced by a particular refractive surgical procedure, such as photorefractive keratectomy or laser in situ keratomileusis. Continuing efforts aim to identify pharmacological agents that effectively inhibit keratocyte apoptosis without promoting keratocyte necrosis. PMID- 10832716 TI - 8(S)-hydroxyeicosatetraenoic acid is the lipoxygenase metabolite of arachidonic acid that regulates epithelial cell migration in the rat cornea. AB - BACKGROUND: We previously found that the inhibition of lipoxygenases resulted in delayed epithelial wound closure in organ-cultured rat corneas. The present study was undertaken to determine the lipoxygenase enzyme and metabolite(s) responsible for regulating reepithelialization and their mechanism of action. METHODS: The effects of esculetin--an established lipoxygenase inhibitor--on endogenous hydroxyeicosatetraenoic acids (HETEs) production, epithelial wound closure, filamentous-actin (F-actin) cytoskeleton, and mitotic rate were investigated using a cell-culture assay and an organ-culture assay of rat corneal epithelium. RESULTS: Lipoxygenase inhibition by esculetin, which resulted in the disruption of F-actin organization and a decrease in the mitotic rate, delayed wound closure in both cell- and organ-culture assays. Normal corneoscleral rims metabolized [3H]arachidonic acid to 12-HETE (major metabolite), 8-HETE, and 9-HETE. HETE synthesis was inhibited by esculetin in a dose-dependent fashion. Chiral-phase analysis revealed that they contained only (S)-enantiomers, which indicated that they were lipoxygenase metabolites. The inhibitory effects of esculetin on F actin organization and epithelial wound closure in an organ-culture assay were totally reversed by exogenously added 8(S)-HETE, whereas 12- and 9-HETE had no effect. However, none of the HETEs reversed the decreased mitotic rate or achieved complete wound closure in the cell-culture assay. CONCLUSIONS: These results suggest that 8(S)-HETE is the key metabolite of arachidonic acid that regulates corneal epithelial cell migration during wound healing. The metabolite responsible for cell proliferation remains to be determined. PMID- 10832717 TI - The spectrum of beta ig-h3 gene mutations in Japanese patients with corneal dystrophy. AB - PURPOSE: This study was undertaken to identify beta ig-h3 gene mutations in Japanese patients with granular corneal dystrophy (GCD), Avellino corneal dystrophy (ACD), lattice corneal dystrophy (LCD), and Reis-Bucklers' corneal dystrophy (RBCD). R124H, R124C, R555W, and R555Q mutations have been reported in Europe to cause ACD, LCD type I, GCD, and RBCD, respectively. METHODS: In total, 91 Japanese patients who had been clinically diagnosed with GCD, LCD, or RBCD were investigated to determine whether they had mutations in the beta ig-h3 gene. Genomic DNA was amplified using the polymerase chain reaction and analyzed using single-strand conformation polymorphism techniques. Mutations were identified using the direct sequencing method. RESULTS: In 68 unrelated patients who had been diagnosed with GCD, 62 patients (91%) were found to have the R124H mutation, which has been reported to cause ACD, whereas only six patients (9%) had the R555W mutation. In LCD patients, 10 patients with type I disease had the R124C mutation, and 10 patients with type IIIA disease had a P501T mutation. One patient with atypical LCD had an L527R mutation. In two patients with RBCD, one had an R555Q mutation and the other patient with geographic opacities was found to have an R124L mutation. CONCLUSIONS: Depending on the specific mutation in the beta ig-h3 gene, the phenotypes of corneal dystrophy may differ. Our results indicate that assay of mutations in the beta ig-h3 gene is required to establish a correct diagnosis. PMID- 10832718 TI - Immunoregulation by aqueous humor. AB - PURPOSE: The eye is a classic example of an immune-privileged site. To investigate the local defense system of the eye, the immunosuppressive effects of the aqueous humor on cytokine production was examined. METHODS: Using T cell clones (TCCs) established from infiltrating cells in the aqueous humor of patients with human T lymphotropic virus type 1 (HTLV-1) uveitis as target cells and the aqueous humor of patients with senile cataract obtained during cataract surgery, we examined the effects of the aqueous humor on the production of cytokines by HTLV-1-infected TCCs. RESULTS: HTLV-1-infected TCCs produced large amounts of various cytokines. The aqueous humor inhibited the production of cytokines in a dose-dependent manner. The inhibitory activity was heat labile. First protein liquid chromatography showed at least four major peaks of different molecular size, indicating that the aqueous humor contains multiple immunosuppressive factors. Transforming growth factor-beta, alpha-melanocyte stimulating hormone, and vasoactive intestinal peptide did not suppress cytokine production. The inhibitory activity was neutralized by monoclonal antibody to Fas ligand. CONCLUSION: These data suggest that soluble Fas ligand is a candidate suppressive factor in the aqueous humor. PMID- 10832719 TI - Establishment of a human lacrimal gland epithelial culture system with in vivo mimicry and its substrate modulation. AB - PURPOSE AND METHODS: Viable single human lacrimal gland (LG) epithelial cells were obtained by serial incubation in chelating and enzymatic solutions. These cells were evaluated for outgrowth in growth factor-enriched medium using the following culture substrates: Matrigel type I collagen gel with or without fibroblasts, and plastic. Each epithelial outgrowth was characterized morphologically and immunohistochemically, and their growth and viability were examined by bromodeoxyuridine (BrdU) labeling and a quantitative cell viability assay. Synthesized proteins were evaluated by enzyme-linked immunosorbent assay (ELISA), sodium dodecyl sulfate polyacrylamide gradient gels, and 14C-amino acid incorporation. RESULTS: LG epithelial cells plated on Matrigel formed clusters with central cavities that contained lactoferrin, mimicking acinar complexes in vivo. Cells plated on Matrigel containing fibroblasts formed tubuloacinar structures and resembled the human LG in vivo. Cells plated on collagen gel or collagen gel containing fibroblasts formed islands or a monolayer, and lactoferrin was detected in incomplete cavities of epithelia on the latter substrate. Epithelial cells plated on plastic formed a monolayer, and cellular expression of lactoferrin was weak and sporadic. Cellular release of lactoferrin measured by ELISA supported the results of immunohistochemistry. Significant differences in proliferative rate were noted between substrates; cells grown on plastic had the highest proliferative rate, whereas cells grown on Matrigel had the lowest rate. CONCLUSION: Culture substrates modulate LG epithelial cell morphology, proliferative rate, and production of the tear protein lactoferrin. Matrigel promotes acinar differentiation to a greater extent than collagen gel and plastic. Incorporation of fibroblasts in substrates promotes ductal differentiation. PMID- 10832720 TI - Reflective meniscometry: a new field of dry eye assessment. AB - PURPOSE: To report a newly developed noninvasive method to measure the radius of tear meniscus curvature (reflective meniscometry) and its application to normal eyes, dry eyes, and other ocular surface conditions. METHODS: Two types of meniscometer--photographic and video--were devised. Both use tear meniscus as a concave mirror, and a specular reflex of an illuminated target at the meniscus was photographed or videotaped to obtain the radius of tear meniscus curvature. The photographic system was applied to 45 normal and 32 dry eyes; in the dry eyes, the radius of tear meniscus curvature was compared to the results of other dry eye examinations. The video system was used to examine meniscus formation on the ocular surface in patients with ocular surface irregularities. RESULTS: The radii of tear meniscus curvature were significantly lower in dry eyes (0.250 +/- 0.086 mm, mean +/- SD) than in normal eyes (0.365 +/- 0.153; p = 0.0003). There were significant correlations between the radius and fluorescein staining score (p = 0.0032) and the grading of interference colors on the precorneal oil film (p = 0.0125). Videomeniscometry demonstrated the tear meniscus to be very stable and proved to be useful in the study of menisci at the edge of rigid contact lenses and other menisci encountered in ocular surface disorders. CONCLUSIONS: Measurement of the radius of tear meniscus curvature may be useful in the diagnosis of dry eyes. Reflective meniscometry is expected to find wide applications in the analysis of tear meniscus in ocular surface disorders. PMID- 10832721 TI - Dry eye and closed eye tears. AB - PURPOSE: To establish accurate measurement of tear production in the diagnosis and monitoring of dry eye. METHODS: Reexamination of the Schirmer test indicated that the rate of tear secretion decreases with advancing age. We measured the noninvasive tear film break-up time (NIBUT) in normal and dry eye patients with our original apparatus. The NIBUT showed a reasonable level of accuracy with a cut-off value of 5 seconds. The lactoferrin concentration in tear fluids subsequently was found to have a cut-off value of 1.1 mg/mL with an optimal level of accuracy. RESULTS: The present findings suggest that a combination of clinical tests is needed for the diagnosis of dry eye syndrome. The closed eye condition induces subclinical inflammation on the ocular surface. Fibronectin, a high molecular weight glycoprotein, plays an important role in corneal wound healing, although its concentration in various types of tear has not been investigated. Fibronectin concentration in open eye tears (19 +/- 24 ng/mL) was significantly different (p = 0.004) from that in closed eye tears (4127 +/- 3222 ng/mL). During reflex tearing with nasal stimulus, the concentration increased significantly (p = 0.028) after 100 mL of reflex tears (220 +/- 126 ng/mL). Albumin concentration in the same samples showed a pattern similar to that for fibronectin. Administration of a topical vasoconstrictor eliminated the increase in fibronectin concentration during reflex tearing. CONCLUSION: These findings suggest that fibronectin in the tear fluid was derived from plasma and that the increase in concentration in closed eye and reflex tear fluid was caused by leakage from dilated conjunctival blood vessels. PMID- 10832722 TI - Helicobacter pylori infection, host genes, and disease outcome. PMID- 10832723 TI - Susceptibility to leukemia and resistance to solid tumors in Down syndrome. PMID- 10832724 TI - Opioid tolerance to sedation and analgesia. PMID- 10832725 TI - A new approach to measure energy expenditure in the neonate. PMID- 10832726 TI - From prevention to prognosis: clinical research update on adolescent eating disorders. PMID- 10832727 TI - Hippocampal volume and everyday memory in children of very low birth weight. AB - Children born preterm and of very low birth weight have an increased incidence of learning difficulties, but little is known about the specific nature of their cognitive deficits and the underlying neuropathology. We hypothesized that their vulnerability to hypoxic, metabolic, and nutritional insults would lead to reduced hippocampal volumes and to deficits in memory because of the role of the hippocampus in this domain of cognition. Neuropsychological and magnetic resonance imaging methods were used to investigate this hypothesis in adolescents born preterm (< or = 30 wk gestation, n = 11) or full-term (n = 8). The preterm group had significantly smaller hippocampal volumes bilaterally, despite equivalent head size, and showed specific deficits in certain aspects of everyday memory, both on objective testing and as indicated by parental questionnaires. The preterm group also had a specific deficit in numeracy. The reduced hippocampal volumes and deficits in everyday memory have previously been unrecognized, but their prevalence in a group of neurologically normal children is striking. PMID- 10832728 TI - The relationship between placental and other perinatal risk factors for neurologic impairment in very low birth weight children. AB - Placental abnormalities reflect antenatal disease processes that may interact with other perinatal risk factors to affect long-term outcome. We performed a nested case control analysis of placental and clinical risk factors associated with neurologic impairment (NI) at 20-mo corrected age (60 cases and 59 controls) using data collected in a prospective study of very low birth weight (less than 1500 g) infants born between 1983 and 1991. In a preliminary analysis we explored the relationship between clinical infection and histologic chorioamnionitis (CA). Only histologic CA with a fetal vascular response correlated with either clinical CA or early onset neonatal sepsis. We then assessed the relative contribution of the nine risk factors (four placental and five clinical) associated with NI at the univariate level by multiple logistic regression. Three risk factors were independent predictors of NI: severe cranial ultrasound abnormalities (odds ratio 13.6, 95% confidence intervals 4.5-66.7), multiple placental lesions (odds ratio 13.2, 95% confidence intervals 1.3-137.0), and oxygen dependence at 36 wk (odds ratio 4.2, 95% confidence intervals 1.2-14.6). Finally, a series of logistic regressions was conducted with the dependent variable changing as we moved back along the causal chain to explore the relationships between risk factors operating at different stages. This analysis suggested that antenatal variables that were not independent predictors of NI by multiple logistic regression exerted their effects through the following intermediate pathways: fetal grade 3 histologic CA via chorionic vessel thrombi, clinical CA via grade 3 villous edema, and grade 3 villous edema via severe cranial ultrasound abnormalities. PMID- 10832729 TI - Sedative tolerance accompanies tolerance to the analgesic effects of fentanyl in infant rats. AB - Iatrogenic tolerance and physical dependence have been documented in human neonates and infants infused with fentanyl or morphine i.v. to maintain continuous analgesia and sedation during extracorporeal membrane oxygenation (ECMO) and mechanical ventilation for the treatment of life-threatening pulmonary diseases. Using postnatal d 17 infant rats, the hypothesis was tested that sedative tolerance accompanies tolerance to fentanyl analgesia in the tail-flick test. Postnatal d 14 infant rats remained naive or received osmotic minipumps infusing saline (1 microL/h) or fentanyl citrate (60 microg x kg(-1) h(-1)). Seventy-two hours later, fentanyl's antinociceptive potency was reduced 3.1-fold in fentanyl-infused rats. Conscious sedation and deep sedation were examined with the cliff-avoidance and the righting-reflex procedures, respectively. Fentanyl infused infants were tolerant to both the conscious and deep sedative effects of fentanyl. Another hypothesis tested was that very high receptor intrinsic activity opioids are less likely to produce tolerance, or to be cross-tolerant to other opioids. Dihydroetorphine is 5,000 to 10,000 times more potent than morphine. However, fentanyl-infused infant rats were cross-tolerant to the analgesic and sedative effects of dihydroetorphine. Interestingly, dihydroetorphine's analgesic efficacy was significantly reduced to a maximum analgesic efficacy (Emax) value of 40% maximum possible effect (MPE). Another concern was whether fentanyl tolerance would generalize to another class of sedatives, the benzodiazepines. This was especially relevant considering the widespread use of benzodiazepines like midazolam in ECMO and mechanical ventilation. Midazolam elicited no analgesia in the tail-flick test. Furthermore, fentanyl-tolerant rats were not cross-tolerant to the conscious or deep sedative effects of midazolam. PMID- 10832730 TI - Intrauterine infection induces programmed cell death in rabbit periventricular white matter. AB - An association between chorioamnionitis and periventricular leukomalacia has been reported in human preterm infants. However, whether this link is causal has not been convincingly established, and the underlying molecular mechanisms remain unclear. The objective of this study was to establish a reproducible model of cerebral white matter disease in preterm rabbits after intrauterine infection. Escherichia coli was inoculated into both uterine horns of laparotomized pregnant rabbits when gestation was 80% complete. The fetuses were delivered by cesarean section and killed 12, 24, or 48 h after the inoculation. Programmed cell death in the white matter was evaluated by hematoxylin-eosin-saffron staining and in situ fragmented DNA labeling (terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling). In a first group of 14 pregnant rabbits not treated with antibiotics, all fetuses delivered 48 h after inoculation were stillborn, whereas fetuses extracted 12 or 24 h after inoculation were alive. No significant cell death was detected in the live fetuses compared with the control noninfected rabbits. In a second group of five pregnant rabbits treated with ceftriaxone initiated 24 h after the inoculation and continued until cesarean section was performed 48 h after inoculation, 13 fetuses were alive, but all showed evidence of extensive programmed cell death in the white matter by hematoxylin-eosin saffron staining and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling. White matter damage became histologically detectable only 48 h after inoculation. Three of the 13 brains displayed periventricular white matter cysts mimicking human cystic periventricular leukomalacia. The high reproducibility of white matter damage in our model should permit further studies aimed at unraveling the molecular mechanisms of periventricular leukomalacia. PMID- 10832731 TI - Enzyme replacement therapy in a feline model of MPS VI: modification of enzyme structure and dose frequency. AB - Enzyme replacement therapy (ERT) in the MPS VI cat is effective at reducing or eliminating pathology in most connective tissues. One exception is that cartilage and chondrocytes remained distended with extensive lysosomal vacuolation after long-term, high-dose ERT. In this study, we demonstrate that recombinant human N acetylgalactosamine-4-sulphatase (4S) is taken up by chondrocytes via a mannose-6 phosphate-dependent mechanism and is effective at removing MPS storage. In vitro, the penetration of 4S into articular cartilage is low (partitioning coefficient = 0.06) and i.v. administered enzyme does not distribute significantly into articular cartilage in vivo. To alter the tissue distribution of 4S, the enzyme was coupled to ethylene diamine or poly-L-lysine, increasing its overall charge and diffusion into cartilage, and the dosing frequency of unmodified 4S was increased. Modification resulted in active 4S that maintained its ability to correct MPS storage and increased the partitioning coefficient of 4S into cartilage by 77% and 50% for ethylene diamine and poly-L-lysine, respectively. However, in vivo ERT studies demonstrated that response to therapy was not significantly improved by either the enzyme modifications or change to the dosing regimen, when compared with ERT with unmodified enzyme. Distribution experiments indicated the majority of enzyme is taken up by the liver irrespective of modification. To optimize therapy and improve the amount of enzyme reaching cartilage and other tissues demonstrating poor uptake, it may be necessary to bypass the liver or prolong plasma half-life so that proportionately more enzyme is delivered to other tissues. PMID- 10832732 TI - Mucopolysaccharidosis type VII in the developing mouse fetus. AB - Mucopolysaccharidosis type VII (MPS VII) is a lysosomal storage disease caused by a deficiency of beta-glucuronidase (1). MPS VII is a fatal, progressive degenerative disorder, and a number of patients die of hydrops fetalis. Thus an approach to treating this disease may be by transplantation or gene therapy in utero. A mouse model of MPS VII has been studied extensively but the disease in affected fetal mice has not been characterized, which is essential for evaluation of therapeutic efficacy. Fetal and newborn mice affected with MPS VII were examined for lysosomal enzyme activities and for the presence of typical storage lesions in comparison to normal and carrier littermates. No beta-glucuronidase enzymatic activity was detected in any of the tissues of affected mice, indicating that transplacental transfer of beta-glucuronidase from the dam did not occur. Lesions were not detected in affected fetuses of 13.5 d gestational age on light or electron microscopy. Vacuolation in cells, typical of lysosomal accumulation of substrate, was first seen in a small number of cells of the reticulo-endothelial system in 15.5 d gestational age livers and in 18.5 d gestational age brains. Storage lesions were not seen consistently in endothelial and Kupffer cells of fetal livers until 18.5 d gestational age and in brains until birth. The results suggest that treatment of affected mice performed at 13.5 d gestational age may be effective in forestalling disease manifestations. PMID- 10832733 TI - Abnormal immune function in vivo in a murine model of lysosomal storage disease. AB - Lysosomal storage diseases are a class of inborn errors of metabolism that lead to widespread disease in multiple tissues. The murine model of mucopolysaccharidosis type VII (MPS VII) closely parallels the human syndrome and has been extensively used to investigate the natural history and therapeutic strategies for lysosomal storage diseases in general. Here we demonstrate a previously undescribed immune defect in the MPS VII mouse. Although the normal populations of cells are present in lymph nodes of these mice, MPS VII mice show a blunted T cell proliferative response and decreased antibody production after immunization with antigens. One mechanism of this defect is ineffective processing of protein antigens, as responses to peptide antigens are normal. This phenotype is presumably caused by the lysosomal disorder, as the defect can be corrected in vivo by direct enzyme replacement therapy. These findings have implications for the use of this animal model, and may have clinical significance for other, more-common lysosomal storage diseases. PMID- 10832734 TI - Outcome of pediatric thromboembolic disease: a report from the Canadian Childhood Thrombophilia Registry. AB - The outcome for children with deep vein thrombosis (DVT) and pulmonary embolism (PE) is unknown. An understanding of morbidity and mortality of DVT/PE is crucial to the development of rational treatment protocols. The Canadian Childhood Thrombophilia Registry has followed 405 children aged 1 mo to 18 y with DVT/PE for a mean of 2.86 y (range, 2 wk to 6 y) to assess outcome. The all-cause mortality was 65 of 405 children (16%). Mortality directly attributable to DVT/PE occurred in nine children (2.2%), all of whom had central venous line-associated thrombosis. Morbidity was substantial, with 33 children (8.1%) having recurrent thrombosis, and 50 children (12.4%) having postphlebitic syndrome. Recurrent thrombosis and postphlebitic syndrome were more common in older children, although deaths occurred equally in all age groups. The incidence of recurrent thrombosis and postphlebitic syndrome are likely underestimated because of difficulties in diagnosis, especially in younger children. The significant mortality and morbidity found in our study supports the need for international multicenter randomized clinical trials to determine optimal prophylactic and therapeutic treatment for children with DVT/PE. PMID- 10832735 TI - Ductus venosus flow velocity in newborn lambs during increased pulmonary artery pressure. AB - The aim of the present study was to assess with ultrasound the ductus venosus flow velocity in newborn lambs with increasing pulmonary artery pressures and to evaluate whether this is a useful method to detect elevated pulmonary artery pressure. The ductus venosus flow velocity was studied with pulsed-wave Doppler echocardiography in nine newborn lambs < or = 30 h old. The lambs were anesthetized, mechanically ventilated, and instrumented to measure mean airway pressure and pulmonary artery and arterial blood pressures. A vascular occluder was placed around the main pulmonary artery. With mean pressures ranging from 20 to 50 mm Hg in the pulmonary artery, the ductus venosus flow velocity was examined. In seven lambs, the mean portal pressure and central venous pressure were also measured. With a stepwise increase in the pulmonary artery pressure, the minimum ductus venosus flow velocity during atrial systole decreased to a reversed flow, and the duration of this reversed flow component increased. The systolic forward peak flow velocity signal also gradually decreased. No changes were detected in the mean central venous or in the portal pressure with increasing pulmonary artery pressure or changes in ductus venosus flow. The flow velocity in the ductus venosus, which is higher than in other precordial veins, shows a reduction and even reversal of the nadir and an increase of the duration of reversed flow during atrial systole as a response to increased pulmonary artery pressure. Thus, Doppler ultrasound of the ductus venosus flow velocity may be a useful noninvasive diagnostic supplement to detect pulmonary hypertension of the newborn. PMID- 10832736 TI - Daily rhythms in renal blood flow and urine production rate in the near-term sheep fetus. AB - Daily rhythmicity of renal blood flow (RBF) and urine flow (UF) was studied in fetal sheep between 121-125 d of gestation. Fetal arterial blood pressure, heart rate, UF, and right RBF were measured continuously for 24-h periods in 10 sheep. Rhythmic variations during a 24-h period were found for all variables studied. The rhythms of arterial blood pressure and heart rate were highly correlated, whereas an inverse correlation was found between arterial blood pressure with RBF and between arterial blood pressure with UF. These findings indicate that fetal RBF is not blood pressure dependent. Furthermore, fetal UF appears not to be mediated by pressure-dependent diuresis. PMID- 10832737 TI - Gender-related heart rate differences in human neonates. AB - The aim of the present study was to examine gender-related differences in heart rate of human neonates controlled for their behavior. Previous studies could not find any difference in male and female fetuses and newborns, although this gender dependent difference clearly exists in children and adults. The heart rate of 99 newborns (47 girls and 52 boys) was measured with simultaneous video recording of their behavior. Results proved that alert newborns showed the same difference as adults: boys had a significantly lower baseline heart rate than girls. This suggests that heart rate is gender-dependent from birth onward. PMID- 10832738 TI - Effect of maternal nutrition on brown adipose tissue and its prolactin receptor status in the fetal lamb. AB - We investigated the influence of maternal nutritional enhancement during the second half of gestation on prolactin receptor (PRLR) abundance in fetal brown adipose tissue (BAT) and liver close to term (i.e. 141-144 d gestation). Ewes were provided with 100% (i.e. control; n = 8) or 150% (i.e. well-fed; n = 7) of their metabolic requirements from 80 to 144 d gestation. Crude plasma membranes were prepared from fetal BAT and hepatic tissue, and individual molecular weight isoforms for the long and short forms of the PRLR were detected by immunoblotting. Mitochondrial preparations were prepared from BAT to measure the amount of the BAT-specific mitochondrial uncoupling protein-1 and its thermogenic activity (i.e. guanosine 5'-diphosphate binding). Fetuses sampled from well-fed ewes were heavier (controls, 3927 +/- 196 g; well-fed, 4783 +/- 219 g; p = 0.01) but possessed less BAT per kilogram body weight (controls, 5.92 +/- 0.43 g/kg; well-fed, 3.85 +/- 0.19 g/kg; p = 0.001), which had a greater uncoupling protein 1 abundance (controls, 56 +/- 5% of reference; well-fed, 78 +/- 9% of reference; p < 0.01) and higher thermogenic activity (controls, 157 +/- 41 pmol guanosine 5' diphosphate per milligram mitochondrial protein; well-fed, 352 +/- 36 pmol guanosine 5'-diphosphate per milligram mitochondrial protein; p < 0.01) than controls. Multiple isoforms of the long and short forms of the P1LR were detected in all tissues. BAT from well-fed fetuses had a higher abundance of the 15-kD isoform of the long form of the PRLR (controls, 1.6 +/- 0.4 densitometric units; well-fed, 16.3 +/- 2.0 densitometric units; p < 0.001). This isoform was not detected in hepatic tissue. Maternal nutrient intake had no effect on any other isoforms of the PRLR in BAT or liver. In conclusion, increasing the quantity of feed provided in late gestation acts to promote fetal weight and BAT maturation, the combination of which will enhance neonatal viability. PMID- 10832739 TI - Validation of a [13C]bicarbonate tracer technique to measure neonatal energy expenditure. AB - The use of a stable isotope-labeled [13C]bicarbonate infusion to measure energy expenditure is advantageous, as a complete collection of expired air is not required. This technique allows for facile measurements of energy expenditure in intubated neonates. The aim of the present study was to determine the accuracy of energy expenditure estimates in postsurgical neonates by using the [13C]bicarbonate method compared with the current standard, indirect calorimetry. Eight neonates who were receiving total parenteral nutrition [98 +/- 21 (SD) kcal x kg(-1) x d(-1); 3.1 +/- 0.7 (SD) protein g x kg(-1) x d(-1)] were studied on postoperative d 15.5 +/- 11.9. A primed continuous 3-h intravenous infusion of NaH13CO3 and indirect calorimetry were performed simultaneously. Energy expenditure was calculated separately from the Weir equation and from the dilution of 13CO2 in the breath in combination with the individual energy equivalents of CO2 from the diet. The rate of CO2 appearance and energy expenditure calculated from the bicarbonate method (0.725 +/- 0.021 mol x kg(-1) x d(-1); 89.5 +/- 2.5 kcal x kg(-1) x d(-1)) highly correlated (r = 0.94 and 0.98, respectively) with the CO2 excretion and energy expenditure determined by indirect calorimetry (0.489 +/- 0.016 mol x kg(-1) x d(-1); 60.2 +/- 2.0 kcal x kg(-1) x d(-1)) when analyzed nonproportionately to weight. Bland-Altman analysis demonstrated the 95% confidence interval to be +/- 8.2 kcal x kg(-1) x d(-1). Linear regression analysis revealed a highly statistically significant equation relating the two energy expenditures: Indircal (kcal/d) = -9.341 + [0.705 x Bicarb (dcal/d)]; p < 0.001, r2 = 96.4%. We conclude that energy expenditure in neonates can be accurately determined using the [13C]bicarbonate method and a regression equation. Therefore, the bicarbonate method may be useful for determining energy expenditure in neonates not readily accessible to indirect calorimetry, such as those being mechanically ventilated or on extracorporeal life support. PMID- 10832740 TI - Uteroplacental insufficiency alters cerebral mitochondrial gene expression and DNA in fetal and juvenile rats. AB - Uteroplacental insufficiency increases the risk of perinatal and long-term neurologic morbidity by depriving the fetus of oxidative substrate and causing intrauterine growth retardation. Skeletal muscle and liver from growth retarded fetal and juvenile rats respond to this deprivation by altering mitochondrial gene expression and function. The objective of this study was to determine whether cerebral mitochondrial mRNA is similarly altered in fetal and juvenile growth retarded rats and to correlate these alterations with mitochondrial DNA and marker protein levels. To fulfill this objective, mRNA levels of four important mitochondrial proteins were quantified using RT-PCR in growth retarded and sham-operated control fetal and juvenile rat brains; these proteins were NADH ubiquinone oxireductase subunit 4, subunit C of the F1F0-ATPase, and the adenine nucleotide transporters 1 and 2. Mitochondrial DNA/nuclear DNA ratios and mitochondrial 60 kD marker protein levels were also quantified in growth retarded and sham-operated control fetal and juvenile rat brains using PCR and Western Blotting, respectively. Cerebral mRNA levels of all four proteins were increased in the IUGR fetuses and decreased in the IUGR juvenile animals. Cerebral mitochondrial/nuclear DNA ratios and mitochondrial marker protein levels were not significantly altered in the IUGR fetuses; however, both were significantly diminished in IUGR juvenile pups. These studies suggest that the metabolic stresses associated with uteroplacental insufficiency in the rat cause altered fetal and postnatal cerebral mitochondrial mRNA and DNA levels. PMID- 10832741 TI - Immunohistochemical localization of protein 3-nitrotyrosine and S-nitrosocysteine in a murine model of inhaled nitric oxide therapy. AB - Inhaled nitric oxide (INO) therapy is currently used clinically to selectively dilate the pulmonary vasculature and to help treat persistent pulmonary hypertension and bronchopulmonary dysplasia in the neonate. However, in the presence of oxygen or superoxide, nitric oxide forms potentially harmful reactive nitrogen species. Using an experimental mice model, we examined the effects of concurrent hyperoxia and INO on protein tyrosine nitration and cysteine S nitrosylation in pulmonary tissue. Data showed enhanced 3-nitrotyrosine staining within the airway epithelium and alveolar interstitium of mice lungs treated with hyperoxia, which did not increase significantly with INO administration. Within the alveolar interstitium, 3-nitrotyrosine staining was localized to macrophages. S-Nitrosocysteine staining in airway epithelium was significantly enhanced with INO administration regardless of oxygen content. These data suggest that the formation of protein S-nitrosocysteine is the major protein modification during administration of INO. PMID- 10832742 TI - Effect of increased lung expansion on lung growth and development near midgestation in fetal sheep. AB - Obstruction of the fetal trachea is a potent stimulus for fetal lung growth and may have therapeutic potential in human fetuses with lung hypoplasia. However, the effects of increased lung expansion on lung development near midgestation, which is the preferred timing for fetal intervention, have not been well studied. Our aim was to determine the effects of increased lung expansion on lung development at 75-90 d of gestation in fetal sheep. In three groups of fetuses (n = 4 for each), the trachea was occluded for either 10 [10-d tracheal occlusion (TO) group] or 15 d (15-d TO group) or left intact (control fetuses). TO for both 10 and 15 d caused fetal hydrops, resulting in significantly increased fetal body weights. Both periods of TO significantly increased total lung DNA contents from 99.8 +/- 10.1 to 246.0 +/- 5.3 and 246.9 +/- 48.7 mg in 10- and 15-d TO fetuses, respectively. TO for 10 and 15 d also increased airspace diameter, although the percentage of lung occupied by airspace was not increased in 10-d TO fetuses due to large increases in interairway distances; this resulted from a large increase in mesenchymal tissue. The interairway distances at 15 d of TO were reduced compared with the 10-d value but were still approximately 30% larger than control values. We conclude that TO at <90 d of gestation in fetal sheep induces a greater increase in lung tissue growth than later in gestation but also causes fetal hydrops and produces changes in lung structure that are not compatible with efficient gas exchange. Thus, increased lung expansion at a similar stage of development in human fetuses is unlikely to induce changes in lung development that would facilitate gas exchange after birth. PMID- 10832743 TI - Variability of the breathing pattern in newborn rats: effects of ambient temperature in normoxia or hypoxia. AB - We hypothesized that the inter-breath variability of the breathing pattern in newborn rats varied with temperature and oxygenation. Breathing pattern was recorded in 4-day-old rats by airflow plethysmography, during normoxia in warm (control) and cold conditions, or during hypoxia (inspired O2 = 10%) in warm or cold conditions, each lasting 15 min. The warm phase (36 degrees C) either preceded or followed the cold (24 degrees C). Time-domain analysis was applied to 500 continuous breaths recorded toward the end of each phase. All parameters describing the breathing pattern (instantaneous ventilation, tidal volume, and inspiratory and expiratory time) had lower variability when the condition differed from control i.e. in cold or hypoxia, with no correlation with the absolute level of ventilation. The difference in variability between warm normoxia and the other conditions was reduced when cold preceded the warm phase. Gaseous metabolism was increased in cold because of thermogenesis. When the cold preceded the warm phase the increased thermogenesis partly persisted into the warm phase, raising the metabolic level. We conclude that the variability of the breathing pattern in newborn rats 1) does not depend on the absolute level of ventilation, and 2) is reduced by the increased chemical stimuli occurring during cold-hypermetabolism or hypoxia. In normoxia in warm condition metabolic and chemical stimuli are low, and the variability is the highest. The results are in agreement with the clinical observations of a higher incidence of apneic episodes in infants during warm conditions. PMID- 10832744 TI - Effect of ICAM-1 blockade on lung inflammation and physiology during acute viral bronchiolitis in rats. AB - Viral respiratory infections cause acute bronchiolitis and physiologic dysfunction in human infants and in animals. It is possible that the pulmonary dysfunction is a consequence of the inflammatory cells that are recruited during viral illness. We hypothesized that blockade of intercellular adhesion molecule-1 (ICAM-1), a major cell adhesion molecule, would impede the ingress of leukocytes during viral infection and attenuate virus-induced pulmonary dysfunction. Adult male rats were inoculated with parainfluenza type 1 (Sendai) virus or sterile vehicle, and treated with blocking or nonblocking MAb specific for rat ICAM-1. Respiratory system resistance, oxygenation (PaO2), methacholine responsiveness, and bronchoalveolar lavage (BAL) leukocyte counts were measured in anesthetized, paralyzed, ventilated rats. Treatment with the blocking ICAM-1 antibody reduced virus-induced increases in BAL neutrophils and lymphocytes by 70% (p < 0.001), but did not affect BAL monocytes/macrophages. Peripheral blood leukocyte counts were elevated in anti-ICAM-1 blocking antibody-treated rats (p = 0.0003). Although virus-induced increases in resistance and decreases in PaO2 were not affected by anti-ICAM-1 treatment, there was a small but significant attenuation of virus-induced methacholine hyperresponsiveness (p = 0.02). We conclude that ICAM-1 has an important role in neutrophil and lymphocyte infiltration during respiratory viral illness, and that virus-induced changes in pulmonary physiology are not related directly to the numbers of neutrophils and lymphocytes that migrate to the air spaces during infection. PMID- 10832745 TI - Immunophenotypic changes of fetal cord blood hematopoietic progenitor cells during gestation. AB - We measured cell surface expression of CD34, HLA-DR, CD38, CD19, CD33, CD71, and CD45 antigens in the hematopoietic progenitor cells of fetal cord blood to investigate immunophenotypic changes at different gestational ages. These antigens were identified by flow cytometry in 11 fetuses (gestational age 19-24 wk, in 12 preterm (25-28 wk) and in ten newborn infants born at term. The frequency and number of CD34+ cells were higher in the blood of the 11 fetuses; in addition, a statistically significant inverse correlation between number of CD34+ cells and advancing gestational age was noted. The numbers of CD34+ CD19+, CD34+ CD33+, and CD34+ CD45+ coexpressing cells were significantly higher in the fetuses, whereas CD34+ CD38+ cells were more represented in the neonates at term. Gestational age was inversely correlated with the number of CD34+ CD19+ and CD34+ CD33+ coexpressing cells. A positive correlation between gestational age and CD34+ CD38+ cells was noted. The number of CD34- CD19+, CD34- CD38+, and CD34- CD45+ cells was higher in term infants; furthermore, a significant correlation between advancing gestational age and CD34- CD38+ or CD34- CD45+ cells was demonstrated. The proliferative capacity was also higher at lower gestational ages. These data suggest that the development and lineage commitment of fetal cord blood hematopoietic progenitor cells are very active during the last two trimesters of pregnancy. The most significant changes of hematopoietic cells maturation seem to occur within 25 wk of gestation. PMID- 10832747 TI - Effect of Mycoplasma bovis and Mycoplasma bovigenitalium in semen on fertilization and association with in vitro produced morula and blastocyst stage embryos. AB - Frozen-thawed bovine semen contaminated with Mycoplasma bovis (M. bovis) or Mycoplasma bovigenitalium (M. bovigenitalium) at either a high (10(6) CFU/mL) or low (10(4) CFU/mL) concentration was used for bovine oocyte insemination. The resulting embryos were washed 10 times as recommended by the International Embryo Transfer Society (IETS) prior to isolation of agent. A total of 1494 oocytes was inseminated with contaminated sperm cells and 855 oocytes with uninfected control semen. There was a significantly higher proportion of embryos that developed to the blastocyst stage in control than in the mycoplasma exposed groups (P<0.05). Isolation of motile spermatozoa by swim-up procedure prior to insemination did not render sperm cells free of Mycoplasma spp. Although M. bovis was isolated from all washed embryos after the high exposure level, it was found in only 60% of the samples after the low exposure level. In contrast, M. bovigenitalium was isolated from 70 and 12% of washed embryos exposed to the high and low levels of microorganism, respectively. Using scanning electron microscopy, both microorganisms were detected in association with the surface of zona pellucida intact embryos and with sperm cells. These results indicate that mycoplasmas present in semen can be transmitted through the IVF system and infect embryos. Furthermore, the experiments showed that supplementation of culture media with standard antibiotics and washing embryos as recommended by IETS were not effective in rendering IVF embryos free from M. bovis and M. bovigenitalium. PMID- 10832746 TI - 2-Methylbutyryl-coenzyme A dehydrogenase deficiency: a new inborn error of L isoleucine metabolism. AB - An 4-mo-old male was found to have an isolated increase in 2-methylbutyrylglycine (2-MBG) and 2-methylbutyrylcamitine (2-MBC) in physiologic fluids. In vitro oxidation studies in cultured fibroblasts using 13C- and 14C-labeled branched chain amino acids indicated an isolated block in 2-methylbutyryl-CoA dehydrogenase (2-MBCDase). Western blotting revealed absence of 2-MBCDase protein in fibroblast extracts; DNA sequencing identified a single 778 C>T substitution in the 2-MBCDase coding region (778 C>T), substituting phenylalanine for leucine at amino acid 222 (L222F) and absence of enzyme activity for the 2-MBCDase protein expressed in Escherichia coli. Prenatal diagnosis in a subsequent pregnancy suggested an affected female fetus, supporting an autosomal recessive mode of inheritance. These data confirm the first documented case of isolated 2 MBCDase deficiency in humans. PMID- 10832748 TI - Effects of pre- and post-thaw thermal insults on viability characteristics of cryopreserved bovine semen. AB - The magnitude of damage to the viability of cryopreserved bovine spermatozoa by pre- and post-thaw thermal insults was compared. Semen collected by artificial vagina from 5 Holstein bulls was diluted in egg yolk-citrate-7% glycerol extender (EYCG) and cryopreserved in 0.5 mL French straws at a sperm concentration of 40 to 60 x 10(6) cells/mL. In Experiment 1, straws were subjected to 22, 5 or -18 degrees C static air temperature for a duration of 1, 2, 3, 4 or 5 min before or after thawing in a 37 degrees C water bath for 1 min. Control straws were thawed in a 37 degrees C water bath for 1 min without further thermal insult. In Experiment 2, straws were thawed for 1 min in a 37 (control), 20 or 5 degrees C water bath, or were loaded into an insemination gun and plunged into a 37 degrees C water bath for 3 min. In both experiments, straws were returned to a 37 degrees C water bath for incubation prior to viability analysis. Viability evaluations, conducted in triplicate, included the percentage of motile spermatozoa at 1 min and at 3 h post thermal insult and the percentage of intact acrosomal membranes at 3 h post thermal insult. In both experiments, acrosomal integrity was more sensitive than motility to thermal insult. In Experiment 1, a significant interaction was observed between timing of thermal insult (pre- or post-thaw), static air temperature and duration of straw exposure. At 22 and 5 degrees C, thermal insults applied before thawing significantly (P<0.05) reduced acrosomal integrity at > or = 2 and > or = 4 min of exposure, respectively. However, post thaw exposure to 22 and 5 degrees C for up to 5 min had no effect on any of the sperm viability parameters evaluated. In contrast, at -18 degrees C static air temperature, post-thaw exposure for > or = 3 min decreased acrosomal integrity (P<0.05), while 5 min of pre-thaw exposure was required for alteration of acrosomal integrity. In Experiment 2, each alternative thawing method resulted in significantly (P<0.05) lower incubated acrosomal integrity relative to the controls. These findings suggest that bovine spermatozoa cryopreserved in EYCG extender are more sensitive to pre-thaw than post-thaw thermal insults and that acrosomal integrity following 3-h incubation at 37 degrees C is superior to motility evaluations for detection of damage to sperm viability due to thermal insult. PMID- 10832749 TI - Superovulatory response following transvaginal follicle ablation in cattle. AB - A study was designed to compare superovulatory responses in cattle when gonadotropin treatment followed 1 of 3 different treatments to synchronize follicular wave emergence. Animals at unknown stages of the estrous cycle were randomly assigned to 3 groups: ablation of the 2 largest follicles per pair of ovaries (n = 21); ablation of all follicles > or = 5 mm (n = 19); or intramuscular administration of 5 mg estradiol-17beta plus 100 mg progesterone (n = 23). All animals were given a CIDR-B intravaginally at the time of the respective treatments. Gonadotropin treatment, initiated 1 d after follicle ablation or 4 d after estradiol plus progesterone treatment, in the respective groups, consisted of 200 mg of pFSH divided in decreasing doses twice daily over 4 d. Cloprostenol (500 microg) was given at 48 and 60 h after the first pFSH treatment; CIDR-B devices were removed at the time of the second cloprostenol treatment. Ovarian ultrasonography was done on the days of CIDR-B insertion, first gonadotropin treatment, and at 36 and 72 h after CIDR-B removal. Cattle were inseminated twice, at 60 and 72 h after the first injection of cloprostenol. Ovarian and ova/embryo data were collected at slaughter 5, 6 or 7 d after insemination. No differences were detected among groups in the number of follicles > or = 8 mm at the time of first insemination (20.4 +/- 1.7 vs 16.6 +/- 2.0 vs 19.9 +/- 2.3; P > 0.05). At slaughter, no differences were detected among groups in the numbers of CL (23.3 +/- 1.9 vs 17.9 +/- 1.9 vs 20.1 +/- 2.6; P < 0.05), unovulated follicles > or = 8 mm (2.2 +/- 0.5 vs 2.1 +/- 0.3 vs 3.7 +/- 0.9; P < 0.05), ova/embryos (11.0 +/- 1.4 vs 12.2 +/- 1.3 vs 8.5 +/- 1.3; P < 0.05), fertilized ova (9.4 +/- 1.3 vs 10.1 +/- 1.2 vs 7.5 +/- 1.1; P < 0.05) or transferable embryos (8.2 +/- 1.2 vs 8.4 +/- 1.3 vs 6.5 +/- 0.9; P < 0.05). Variation in the numbers of CL (P = 0.1) and unovulated follicles > or = 8 mm (P < 0.01) was lower in the ablation groups than in the steroid-treated group. Results suggest that follicle ablation is as effective as estradiol plus progesterone in synchronizing follicular wave emergence for superstimulation in cattle, and that ablation of the 2 largest follicles is as efficacious as ablating all follicles > or = 5 mm. PMID- 10832750 TI - Quantitative and qualitative analysis of the effectiveness of a mechanical method for the isolation of preantral follicles from ovine ovaries. AB - The preantral follicles are the major source of oocytes and its utilization has been investigated as an important tool to store large numbers of female gametes for further utilization in reproductive programs. The aim of the present study was to perform quantitative and qualitative analyses of the efficacy of a mechanical method for isolating of preantral follicles from the ovaries of fetuses and from nonpregnant and pregnant ewes, using as reference the population of preantral follicles in situ. In the isolation method the ovaries were cut into fragments in the tissue chopper. Then, the suspension was filtered through nylon mesh filters. The number of isolated follicles per ovary was 1655, 4735 and 4770, respectively, for the fetus, nonpregnant ewe and pregnant ewe. The number of in situ preantral follicles per ovary was 32961, 16627 and 17794, respectively, for the fetus, nonpregnant ewe and pregnant ewe. The follicle recovery rate (number of isolated preantral follicles/number of in situ preantral follicles x 100) was higher in adult ewes (26 and 28%, respectively, for nonpregnant and pregnant ewes) than in fetuses (5%). Histological analysis showed that very few preantral follicles (less than 0.26% in situ and 0.46% after the isolation procedure) were degenerated. In conclusion, this study showed that a mechanical method could be used effectively to isolate a large number of intact ovine preantral follicles. In the future, with improvements in culture systems, the isolation of a great number of oocytes enclosed in preantral follicles will make a valuable contribution to the rare breeds and endangered species, agricultural efficiency and basic research in folliculogenesis. PMID- 10832751 TI - Effect of PGF2alpha and 13,14-dihydro-15-keto-PGF2alpha (PGFM) on corpora luteal function in nonpregnant mares. AB - The objective of this study was to determine if the primary circulating metabolite of PGF2alpha, 13,14-dihydro-15-keto-PGF2alpha (PGFM), is biologically active and would induce luteolysis in nonpregnant mares. On Day 9 after ovulation, mares (n = 7/group) were randomly assigned to receive: 1) saline control, 2) 10 mg PGF2alpha or 3) 10 mg PGFM in 5 mL 0.9% sterile saline i.m. On Days 0 through 16, blood was collected for progesterone analysis. In addition, blood was collected immediately prior to treatment, hourly for 6 h, and then at 12 and 24 h after treatment for progesterone and PGFM analysis; PGFM was measured to verify that equivalent amounts of hormone were administered to PGF2alpha- and PGFM-treated mares. Mares were considered to have undergone luteolysis if progesterone decreased to < or = 1.0 ng/mL within 24 h following treatment. Luteolysis was induced in 0/7 control, 7/7 PGF2alpha-treated, and 0/7 PGFM treated mares. There was no difference (P>0.1) in the occurrence of luteolysis in control and PGFM-treated mares. More (P<0.001) PGF2alpha-treated mares underwent luteolysis than control or PGFM-treated mares. There was no difference (P>0.1) in progesterone concentrations between control and PGFM-treated mares on Days 10 through 16. Progesterone concentrations were lower (P<0.01) on Days 10 through 14 in PGF2alpha-treated compared with control and PGFM-treated mares. There was no difference (P>0.05) in PGFM concentrations between PGF2alpha- and PGFM-treated mares; PGFM concentrations in both groups were higher (P<0.001) than in control mares. These results do not support the hypothesis that PGFM is biologically active in the mare, since there was no difference in corpora luteal function between PGFM-treated and control mares. PMID- 10832752 TI - Activation of bovine oocytes following intracytoplasmic sperm injection (ICSI). AB - In the human and the mouse, intracytoplasmic sperm injection (ICSI) apparently triggers normal fertilization and may result in offspring. In the bovine, injection of spermatozoa must be accompanied by artificial methods of oocyte activation in order to achieve normal fertilization events (e.g., pronuclear formation). In this study, different methods of oocyte activation were tested following ICSI of in vitro-matured bovine oocytes. Bovine oocytes were centrifuged to facilitate sperm injection, and spermatozoa were pretreated with 5 mM dithiothreitol (DTT) to promote decondensation. Sperm-injected or sham injected oocytes were activated with 5 microM ionomycin (A23187). Three hours after activation, oocytes with second polar bodies were selected and treated with 1.9 mM 6-dimethylaminopurine (DMAP). The cleavage rate of sperm-injected oocytes treated with ionomycin and DMAP was higher than with ionomycin alone (62 vs 27%, P < or = 0.05). Blastocysts (2 of 41 cleaved) were obtained only from the sperm injected, ionomycin + DMAP-treated oocytes. Upon examination 16 h after ICSI, pronuclear formation was observed in 33 of 47 (70%) DMAP-treated oocytes. Two pronuclei were present in 18 of 33 (55%), while 1 and 3 pronuclei were seen in 8 of 33 (24%) and 7 of 33 (21%) oocytes, respectively. In sham-injected oocytes, pronuclear formation was observed in 15 of 38 (39%) with 9 (60%) having 2 pronuclei. Asa single calcium stimulation was insufficient and DMAP treatment could result in triploidy, activation by multiple calcium stimulations was tested. Three calcium stimulations (5 microM ionomycin) were given at 30-min intervals following ICSI. Two pronuclei were found in 12 of 41 (29%) injected oocytes. Increasing the concentration of ionomycin from 5 to 50 microM resulted in a higher rate of activation (41 vs 26%). The rate of metaphase III arrest was lower while the rate of pronuclear formation and cleavage development was higher in sperm-injected than sham-injected oocytes, suggesting that spermatozoa contribute to the activation process. Further improvements in oocyte activation following ICSI in the bovine are necessary. PMID- 10832753 TI - Characteristics and seasonal variations in the semen of Alpine, Saanen and Damascus goat bucks born and raised in Greece. AB - Twenty-three purebred Alpine (n=8), Saanen (n=7) and Damascus (n=8) goat bucks raised at the Institute of Reproduction and Artificial Insemination in Thessaloniki, Greece (40 degrees 37 minutes N, 22 degrees 58 minutes E and altitude 32 m above sea level), were used to study the effect of photoperiod on semen production. Samples were collected with an artificial vagina and examined immediately after collection. In spite of the variation in nearly all semen characteristics studied among the 3 breeds of bucks, there was significant seasonal variation in both semen quantity (volume, concentration and total number of spermatozoa per ejaculate) and quality (percentage of motile spermatozoa, percentage of abnormal spermatozoa and rate of progressive motility). The best semen was produced during the breeding season (late summer and autumn). However, the magnitude of these seasonal effects was not sufficient to prevent bucks from being used for breeding throughout the year. Nevertheless, individual differences in the semen quantity and quality among bucks within a breed make individual evaluation of semen necessary to select the most fertile males for breeding. PMID- 10832754 TI - Vitrification of buffalo (Bubalus bubalis) oocytes. AB - The objective of the present study was to develop a method for the cryopreservation of buffalo oocytes by vitrification. Cumulus-oocyte complexes (COCs) were obtained from slaughterhouse ovaries. Prior to vitrification of COCs in the vitrification solution (VS) consisting of 4.5 M ethylene glycol, 3.4 M dimethyl sulfoxide, 5.56 mM glucose, 0.33 mM sodium pyruvate and 0.4% w/v bovine serum albumin in Dulbecco's phosphate buffered saline (DPBS), the COCs were exposed to the equilibration solution (50% VS v/v in DPBS) for 1 or 3 min at room temperature (25 to 30 degrees C). The COCs were then placed in 15-microL of VS and immediately loaded into 0.25-mL French straws, each containing 150 microL of 0.5 M sucrose in DPBS. The straws were placed in liquid nitrogen (LN2) vapor for 2 min, plunged and stored in LN2 for at least 7 d. The straws were thawed in warm water at 28 degrees C for 20 sec. For dilution, the COCs were equilibrated in 0.5 M sucrose in DPBS for 5 min and then washed 4 to 5 times in the washing medium (TCM-199+10% estrus buffalo serum). The proportion of oocytes recovered in a morphologically normal form was significantly higher (98 and 88%, respectively; P<0.05), and the proportion of oocytes recovered in a damaged form was significantly lower (2 and 12%, respectively; P<0.05) for the 3-min equilibration than for 1 min. For examining the in vitro developmental potential of vitrified warmed oocytes, the oocytes were placed in 50-microL droplets (10 to 15 oocytes per droplet) of maturation medium (TCM-199+15% FBS+5 microg/mL FSH-P), covered with paraffin oil in a 35-mm Petri dish and cultured for 26 h in a CO2 incubator (5% CO2 in air) at 38.5 degrees C. Although the nuclear maturation rate did not differ between the 1- and 3-min equilibration periods (21.5+/-10.7 and 31.5+/ 1.5%, respectively), the between-trial variation was very high for the 1-min period. This method of vitrification is simple and rapid, and can be useful for cryopreservation of buffalo oocytes. PMID- 10832755 TI - Effect of body condition on reproductive efficiency of lactating dairy cows receiving a timed insemination. AB - Body condition may influence pregnancy rates to a timed insemination (Ovsynch/TAI) protocol and affect the economical performance of dairy farms. The objectives were to compare pregnancy rates using the Ovsynch/TAI protocol for the first service of lactating dairy cows with body condition scores < 2.5 (scale: 1 to 5, low BCS group) versus > or = 2.5 (control group) and to estimate the economic impact of the effect of body condition on reproductive performance. At 63 +/- 3 d post partum, cows were assigned to 2 experimental groups (low BCS = 81; control = 126), and were treated with GnRH at d 0 and with PGF2alpha 7 d later. At 48 h after PGF2alpha, cows received an injection of GnRH and were inseminated 16 h later. Pregnancy rates to the Ovsynch/TAI protocol were lower for the low BCS group than for the control group at 27 d (18.1 +/- 6.1% < 33.8 +/ 4.5%; P<0.02) and at 45 d (11.1 +/- 5.4% < 25.6 +/- 4.1%; P<0.02) after insemination. Economic analysis indicated that reducing the percentage of the herd in low body condition increases net revenues per cow per year. Body condition influenced pregnancy rates to the Ovsynch/TAI protocol. PMID- 10832756 TI - Outcomes of breeding soundness evaluation of 2898 yearling bulls subjected to different classification systems. AB - This study is a retrospective analysis comparing data on yearling bull breeding soundness evaluation (BSE) subjected to 3 different classification systems: the Society for Theriogenology (SFT) 1983 and 1993 systems, and the Western Canadian Association of Bovine Practitioners (WC) 1993 system. Data were collected at 5 performance bull-test stations located in South Carolina and Tennessee from 1986 through 1996. Yearling bulls (n=2898) that were 10 to 20 mo of age were used in the analysis. To simplify analysis, bulls were determined to be either satisfactory or unsatisfactory potential breeders (including those classified as questionable, deferred or unsatisfactory). Data were analyzed 1) within location where a location was treated as an individual experiment, 2) with breeds and locations collapsed, and 3) within age-group where each age-group was treated as an individual experiment. An ANOVA was performed using GLM procedures of SAS, and this model was used to generate least square means for the proportion of bulls classified as satisfactory and the 5 possible unsatisfactory outcomes due to inadequacies in scrotal circumference, spermatozoal morphology, spermatozoal motility, a combination of inadequate spermatozoal morphology and motility, or physical abnormalities. Inadequate scrotal circumference or physical abnormality did not affect differences for BSE outcomes among systems. Using the SFT93 system, bulls failed at a higher rate due to inadequate spermatozoal morphology (P < 0.05) than when subjected to the SFT83 system. In the WC93 system, a higher percentage of bulls failed due to inadequate spermatozoal motility and to a combination of inadequate spermatozoal morphology and motility than in the other 2 systems (P < 0.05). The proportion of bulls in this data failing under the WC93 system appears to be the result of overestimation. PMID- 10832757 TI - Insemination of mares with low numbers of either unsexed or sexed spermatozoa. AB - Two experiments were conducted to determine pregnancy rates in mares inseminated 1) with 5, 25 and 500 x 10(6) progressively motile spermatozoa (pms), or 2) with 25 x 10(6) sex-sorted cells. In Experiment 1, mares were assigned to 1 of 3 treatments: Group 1 (n=20) was inseminated into the uterine body with 500 x 10(6) pms. Group 2 (n=21) and Group 3 (n=20) were inseminated into the tip of the uterine horn ipsilateral to the preovulatory follicle with 25 and 5 x 10(6) pms, respectively. Mares in all 3 groups were inseminated either 40 (n=32) or 34 h (n=29) after GnRH administration. More mares became pregnant when inseminated with 500 x 10(6) (18/20 = 90%) than with 25 x 10(6) pms (12/21 = 57%; P<0.05), but pregnancy rates were similar for mares inseminated with 25 x 10(6) vs 5 x 10(6) pms (7/20 = 35%) (P>0.1). In Experiment 2, mares were assigned to 1 of 2 treatments: Group A (n=11) was inseminated with 25 x 10(6) spermatozoa sorted into X and Y chromosome-bearing populations in a skimmilk extender. Group B (n=10) mares were inseminated similarly except that spermatozoa were sorted into the skimmilk extender + 4% egg yolk. Inseminations were performed 34 h after GnRH administration. Freshly collected semen was incubated in 224 microM Hoechst 33342 at 400 x 10(6) sperm/mL in HBGM-3 for 1 hr at 35 degrees C and then diluted to 100 x 10(6) sperm/mL for sorting. Sperm were sorted by sex using flow cytometer/cell sorters. Spermatozoa were collected at approximately 900 cells/sec into either the extender alone (Group A) or extender + 4% egg yolk (Group B), centrifuged and suspended to 25 x 10 sperm/mL and immediately inseminated. Pregnancy rates were similar (P>0.1) between the sperm treatments (extender alone = 13/10, 30% vs 4% EY + extender = 5/10, 50%). Based on ultrasonography, fetal sex at 60 to 70 d correlated perfectly with the sex of the sperm inseminated, demonstrating that foals of predetermined sex can be obtained following nonsurgical insemination with sexed spermatozoa. PMID- 10832758 TI - Effect of sterile service on estrus duration, fertility and prolificacy in artificially inseminated dairy goats. AB - The effect of sterile service on estrus duration, fertility and prolificacy in artificially inseminated dairy goats during breeding season was studied. Nubian does (n=126) were divided into 2 equal groups: service and control. Estrus was synchronized with intravaginal sponges containing either fluorgestone acetate (FGA; 40 mg) or medroxiprogesterone acetate (MAP; 60 mg) for 12 or 14 d, respectively. Two vasectomized teaser bucks were used to detect estrus at 6-h intervals for 5 d after sponge removal (0600, 1200, 1800 and 2400 h). The teasers were fitted with aprons and permitted to mount all does in both groups, but to penetrate only the service does within the first 12 h of estrus. Does in both groups were inseminated twice at 12 and 24 h after estrus was first detected, using 1 straw per insemination containing 200 million of cooled spermatozoa from 1 buck. The semen was placed in mid-cervix. Estrus duration for the service and control does was (mean +/- SD) 29.4 +/- 6.5 and 41.8 +/- 9.6 h, respectively. Fertility for the service does was 73.7% (46/63); for control does it was 58.7% (37/63). Prolificacy was 2.1 (96/46) and 2.0 (74/37) for service and control does, respectively. Estrus duration (P<0.001) and fertility (P<0.05) differed between the service and control group, but prolificacy was similar (P>0.05). It is concluded that sterile service reduces the duration of estrus and increases fertility in artificially inseminated dairy goats. PMID- 10832759 TI - Anti-sperm antibodies and seminal characteristics after testicular biopsy or epididymal aspiration in dogs. AB - A study was performed to determine if performing testicular biopsies or epididymal aspirates in dogs would induce sperm-bound anti-sperm antibodies (ASA), affect long-term sperm production or semen quality. Semen was collected from 8 mature dogs 3 times a week before and after hemicastration and then 3 times a week after testicular biopsy (n=3 and 1 control) or epididymal aspiration (n=3 and 1 control). Detection of anti-sperm IgG (ASA) on sperm cells was performed by flow cytometry analysis using a flow cytometer. Two dogs with testicular biopsies became positive for ASA 16 d after testicular biopsy and remained positive for 7 and 9 d, respectively. One dog that had an epididymal aspirate became positive 13 d after epididymal aspiration and remained positive for 35 d. One dog became positive 21 d after hemicastration and remained positive for 28 d. Sperm output declined significantly in 7 of 8 dogs after hemicastration. A control epididymal aspirate treatment dog had decreased sperm output, and a testicular biopsy treatment dog had increased sperm output. None of the dogs with ASA had significant changes in sperm output after treatment. Sperm motility declined significantly in 3 dogs after hemicastration. An epididymal aspiration treatment dog had a decrease in sperm motility, a control epididymal aspirate treatment dog and a control testicular biopsy treatment dog each had increases in sperm motility. None of the dogs with ASA had significant changes in motility. The percentage of normal spermatozoa significantly decreased in 3 dogs and significantly increased in 1 dog after hemicastration. Two dogs that had testicular biopsies and 1 dog that had an epididymal aspiration had decreases in percent normal sperm. Two of 3 dogs with decreases in percent normal sperm after treatment had ASA, but 2 dogs with ASA had no change in motility. Hemicastration, epididymal aspiration, and testicular biopsy can induce ASA production within 2 wk of the procedure, but ASA are transient and do not have a predictably negative effect on total sperm output or motility. Testicular biopsy and epididymal aspiration are safe diagnostic procedures, but further work investigating post treatment fertility must be done before final conclusions can be made. PMID- 10832760 TI - Identification of embryo paternity using polymorphic DNA markers to assess fertilizing capacity of spermatozoa after heterospermic insemination in boars. AB - Differences in sperm fertilizing capacity of males often remain undetected by routine semen parameters. Heterospermic insemination with equal numbers of spermatozoa from 2 males is an accurate method for assessing differences in fertility. Use of heterospermic insemination depends on a reliable, efficient assay to identify paternity of conceptuses or offspring. In this study, polymorphic DNA markers amplified by PCR were tested to determine paternity of Day 5 to 6 embryos. The fertilizing capacity of 2 boars (A and B) with similar semen parameters was compared after homospermic (n=14 gilts) and heterospermic (n=11 gilts) insemination. Single AI's were performed under suboptimal conditions using 1 x 10(9) spermatozoa at 12 to 24 h before ovulation to prompt differences in fertilization and to stimulate sperm competition. The fertilization rate and the number of accessory spermatozoa were determined in Day 5 to 6 embryos. Using 5 different polymorphic DNA markers, paternity could be determined in 95.8% of the embryos. Boar B sired significantly (P<0.05) more offspring than Boar A after insemination with pooled semen, and this was reflected by a significantly (P<0.05) higher number of accessory spermatozoa following homospermic insemination with semen from Boar B, although fertilization rates did not differ between the 2 boars after homospermic insemination. The results suggest that the viability of spermatozoa in the female reproductive tract contributes to differences in fertility rates of males with similar in vitro sperm quality parameters. The number of accessory spermatozoa is a more sensitive measure of boar fertility than the fertilization rate. Polymorphic DNA markers are suitable for verification of parentage even at a very early stage of embryonic development. PMID- 10832761 TI - Periovulatory hormone profiles and components of litter size in gilts with different estrogen receptor (ESR) genotypes. AB - Estrus, endocrine changes during the periovulatory period, and components of litter size at Day 35/36 of pregnancy were studied in gilts with estrogen receptor genotype AA (AA gilts) or BB (BB gilts), in which the B allele is associated with a larger litter size. Neither estrus length nor estrous cycle length was affected by estrogen receptor genotype. No differences in periovulatory plasma LH, estrogen or progesterone profiles between the AA and BB gilts were detected. Furthermore, temporal aspects of these profiles were not different for both genotypes. Although the B allele is associated with a larger litter size, no differences in number of corpora lutea or number and percentage of vital Day 35/36 embryos were found in this study. This indicates that the difference in litter size is not due to differences in oocyte maturation, fertilization, implantation or embryonic survival, but is likely caused by a difference in fetal survival. Thus, uterine capacity might be different for both genotypes. The available uterine space per embryo seems to be the same for both genotypes, as is endometrial folding of uterine surface area. However, a difference in placental size was found. Embryos of BB gilts had significantly longer placentae than embryos of AA gilts. These results suggest a higher chance for placental insufficiency in AA gilts, leading to the expected higher fetal mortality compared with the BB gilts. The difference in placental size might have been related to a difference in the timing of embryonic mortality. PMID- 10832762 TI - Low plasma leptin concentration and low rates of fat oxidation in weight-stable post-obese subjects. AB - OBJECTIVE: A low resting metabolic rate for a given body size and composition, a low rate of fat oxidation, low levels of physical activity, and low plasma leptin concentrations are all risk factors for body weight gain. The aim of the present investigation was to compare resting metabolic rate (RMR), respiratory quotient (RQ), levels of physical activity, and plasma leptin concentrations in eight post obese adults (2 males and 6 females; 48.9 +/- 12.2 years; body mass index [BMI]: 24.5 +/- 1.0 kg/m2; body fat 33 +/- 5%; mean +/- SD) who lost 27.1 +/- 21.3 kg (16 to 79 kg) and had maintained this weight loss for > or =2 months (2 to 9 months) to eight age- and BMI-matched control never-obese subjects (1 male and 7 females; 49.1 +/- 5.2 years; BMI 24.4 +/- 1.0 kg/m2; body fat 33 +/- 7%). RESEARCH METHODS AND PROCEDURES: Following 3 days of weight maintenance diet (50% carbohydrate and 30% fat), RMR and RQ were measured after a 10-hour fast using indirect calorimetry and plasma leptin concentrations were measured using radioimmunoassay. Levels of physical activity were estimated using an accelerometer over a 48-hour period in free living conditions. RESULTS: After adjustment for fat mass and fat-free mass, post-obese subjects had, compared with controls, similar levels of physical activity (4185 +/- 205 vs. 4295 +/- 204 counts) and similar RMR (1383 +/- 268 vs. 1430 +/- 104 kcal/day) but higher RQ (0.86 +/- 0.04 vs. 0.81 +/- 0.03, p < 0.05). Leptin concentration correlated positively with percent body fat (r = 0.57, p < 0.05) and, after adjusting for fat mass and fat-free mass, was lower in post-obese than in control subjects (4.5 +/- 2.1 vs. 11.6 +/- 7.9 ng/mL, p < 0.05). DISCUSSION: The low fat oxidation and low plasma leptin concentrations observed in post-obese individuals may, in part, explain their propensity to relapse. PMID- 10832763 TI - A glucocorticoid receptor gene marker is associated with abdominal obesity, leptin, and dysregulation of the hypothalamic-pituitary-adrenal axis. AB - OBJECTIVE: Abdominal obesity has a key role in the pathogenesis of prevalent and serious diseases and has been shown to be associated with an altered hypothalamic pituitary-adrenal (HPA) axis function, which is regulated by endocrine feedback mediated via hippocampal glucocorticoid receptors (GR). RESEARCH METHODS AND PROCEDURES: We examined the HPA axis function by repeated salivary samples for the assessment of cortisol, as well as other endocrine, anthropometric, metabolic, and circulatory variables in middle-aged Swedish men (n = 284). With the restriction enzyme BclI, variants of the GR gene (GRL) locus were identified and two alleles with fragment lengths of 4.5 and 2.3 kilobases (kb) were detected. RESULTS: The observed frequencies were 40.1% for the 2.3- and 2.3-kb, 46.2% for the 4.5- and 2.3-kb, and 13.7% for the 4.5- and 4.5-kb genotypes. The larger allele (4.5 and 4.5 kb) was associated with elevated body mass index (BMI; p < 0.001), waist-to-hip circumference ratio (p = 0.015), abdominal sagittal diameter (p = 0.002), leptin (p < 0.001), and systolic blood pressure (borderline, p = 0.058). The 4.5- and 4.5-kb allele was associated with leptin after adjustment for BMI. Moreover, salivary cortisol values, particularly after stimulation by a standardized lunch (p = 0.040 to 0.086), were elevated in the men with the larger allele. DISCUSSION: These results indicate that there is an association between a deficient GR function, defined as a poor feedback regulation of the HPA axis activity, and a polymorphic restriction site at the GR gene locus. An abnormal control of HPA axis function due to genetic alterations may contribute to the pathogenesis of abdominal obesity. PMID- 10832764 TI - Sequence variation within the neuropeptide Y gene and obesity in Mexican Americans. AB - OBJECTIVE: Recently, we reported evidence for linkage between neuropeptide Y (NPY) and both obesity and several obesity-related quantitative measures in a sample of Mexican Americans from Starr County, Texas. The purpose of this study was to investigate putative variation within the coding and promoter regions of NPY. RESEARCH METHODS AND PROCEDURES: Five young, obese individuals (body mass index [BMI] 33 to 45 kg/m2, age 14 to 30 years); five adult, lean individuals (BMI 20 to 26 kg/m2, age 39 to 65 years); and five sibling pairs sharing no alleles that were identical by descent at a marker locus proximal to NPY were selected for fluorescence-based sequencing of approximately 1100 base pairs (bp) immediately 5' from the start site and all four exons of NPY. We identified a total of eight variant sites, including a 2-bp insertion/deletion (I/D) within a putative negative regulatory region (-880I/D) and a 17-bp deletion at the exon 1/intron 1 junction (69I/D). The -880I/D and 69I/D variants were typed in a separate random sample of Mexican Americans (N = 914) from Starr County, Texas. RESULTS: Analyses of variance resulted in a significant association between 880I/D and waist-to-hip ratio (p = 0.041) in the entire sample and between 880I/D and BMI (p = 0.031), abdominal circumference (p = 0.044), and waist-to-hip ratio (p = 0.041) in a non-obese subsample (BMI < 30 kg/m2, n = 594). The 69I/D variant was observed in only one pedigree and does not appear to segregate with obesity within this pedigree. DISCUSSION: This study reports newly identified common human sequence variation within the regulatory and coding sequence of NPY. Several variants were observed, and of those tested, the -880I/D promoter region variant may influence body fat patterning in non-obese individuals but does not appear to play a major role in the etiology of common forms of obesity in this population. PMID- 10832765 TI - The effects of drugs used to treat obesity on the autonomic nervous system. AB - OBJECTIVE: Body fatness is partly under hypothalamic control with effector limbs, which include the endocrine system and the autonomic nervous system (ANS). In previous studies we have shown, in both obese and never-obese subjects, that weight increase leads to increased sympathetic and decreased parasympathetic activity, whereas weight decrease leads to decreased sympathetic and increased parasympathetic activity. We now report on the involvement of such ANS mechanisms in the action of anti-obesity drugs, independent of change in weight. RESEARCH METHODS AND PROCEDURES: Normal weight males (ages 22 to 38 years) were fed a solid food diet, carefully measured to maintain body weight, for at least 2 weeks, as inpatients at the Rockefeller University General Clinical Research Center. In a single-blind, placebo/drug/placebo design, eight subjects received dexfenfluramine, seven phentermine (PHE), and seven sibutramine (SIB). ANS measures of parasympathetic and sympathetic activity included: determination of amount of parasympathetic control (PC) and sympathetic control (SC) of heart period (interbeat interval), using sequential pharmacological blockade by intravenous administration of atropine and esmolol. These autonomic controls of heart period are used to estimate the overall level of parasympathetic and sympathetic activities. Norepinephrine, dopamine, and epinephrine levels in 24 hour urine collections were measured and also resting metabolic rate (RMR). RESULTS: Sufficient food intake maintained constant body weight in all groups. PHE and SIB produced significant increases in SC but no change in PC or in RMR. In contrast, dexfenfluramine produced marked decreases in SC, PC, and RMR. For all three drugs, the effects on urine catecholamines directly paralleled changes in cardiac measures of SC. DISCUSSION: ANS responses to PHE and SIB were anticipated. The large, and unanticipated, response to dexfenfluramine suggests further study to determine whether there could be any relation of these ANS changes to the adverse cardiovascular effects of treatment with dexfenfluramine. PMID- 10832766 TI - The relationship of childhood adiposity to parent body mass index and eating behavior. AB - OBJECTIVE: To better understand risk factors for the development of obesity in early childhood, we examined the association between children's adiposity and their parents' eating behavior and body mass index (BMI). RESEARCH METHODS AND PROCEDURES: Parents of 85 white children 36 months of age (49 boys and 36 girls) completed the Three-Factor Eating Questionnaire measuring three dimensions of parent eating behavior: disinhibited eating, cognitive restraint of eating, and susceptibility to hunger. Parent BMI (kg/m2) was calculated using self-reported height and weight. The children's percentage body fat was assessed by dual energy X-ray absorptiometry analysis. RESULTS: Twenty-six percent of parents were obese (BMI > or = 30 kg/m2). Both maternal and paternal BMI were associated with higher scores for disinhibition (r = 0.69 and r = 0.68, p < 0.001), and maternal BMI was also associated with higher scores for hunger (r = 0.51, p < 0.001). There were no significant relationships between children's percentage body fat and parent eating scores, and the correlation between children's percentage body fat and parent BMI was significant only between mothers and daughters (r = 0.35, p = 0.04). Obese parents were no more likely to have a child who was fatter (upper quintile of percentage body fat for gender). DISCUSSION: Among 36 month-old white children, parent eating behavior was related to parent BMI, but not to children's adiposity. There was only a weak relationship between parent BMI and child adiposity. Despite the aggregation of adiposity within families due to shared genes and environments, children may not express differences in susceptibility to obesity by 3 years of age. PMID- 10832767 TI - Care giver perception of children's obesity-related health risk: a study of African American families. AB - OBJECTIVE: To examine care giver perception of children's weight-related health risk in African American families. RESEARCH METHODS AND PROCEDURES: One-hundred and eleven families (representing 48 boys and 63 girls) screened for participation in a diabetes prevention study participated. Care givers completed a health awareness questionnaire that assessed their perception of the child's weight, eating habits, appearance, exercise habits, and health risk. The care givers also reported each subject's family history of obesity, diabetes, and other chronic diseases. After a physical examination, height and weight were used to compute an age- and sex-adjusted body mass index for each child. RESULTS: Despite the fact that a substantial number of children were obese (57%) and super obese (12%), only 44% of the care givers perceived the child's weight to be a potential health problem. Regression analysis showed that 21% of the variance in parental perception of obesity-related health risk could be predicted by child age, body mass index, perception of frame size, and perception of exercise habits. DISCUSSION: A number of reasons for the apparent minimization of child health risk are discussed, including cultural differences in the acceptance of a large body habitus, lack of knowledge about the connection between childhood obesity and future health risk, and an optimistic bias in the perception of personal health risk. PMID- 10832768 TI - Rapamycin inhibits human adipocyte differentiation in primary culture. AB - OBJECTIVE: The immunosuppressant drug rapamycin, has been reported to inhibit 3T3 L1 adipocyte differentiation by interfering with critical postconfluent mitoses that are required early on for successful differentiation of this cell line (clonal expansion phase). In contrast to the murine 3T3-L1 preadipocyte cell line, human preadipocytes in primary culture do not undergo clonal expansion during differentiation. We investigated whether rapamycin could inhibit human adipocyte differentiation. RESEARCH METHODS AND PROCEDURES: The effect of rapamycin on the induction of differentiation of human preadipocytes in primary culture into adipocytes was measured using Oil Red O staining and glycerol phosphate dehydrogenase activity. RESULTS: We have observed that rapamycin severely curtails human adipocyte differentiation of both omental and abdominal subcutaneous preadipocytes (to 14% and 19% of standard differentiation, respectively). The rapamycin-mediated inhibition of human adipocyte differentiation could be reversed in the presence of excess amounts of FK-506, which displaces rapamycin from its intracellular receptor, FKPB12. Measurement of cytosolic protein and [3H]thymidine incorporation into DNA confirmed the absence of proliferation during differentiation of human preadipocytes in primary culture. DISCUSSION: Our data indicate that rapamycin exerts important negative regulatory effects on adipogenesis in human preadipocytes, through a mechanism that does not depend on interruption of clonal expansion. PMID- 10832769 TI - Free fatty acids are involved in the inverse relationship between hormone sensitive lipase (HSL) activity and expression in adipose tissue after high-fat feeding or beta3-adrenergic stimulation. AB - OBJECTIVE: The hormone-sensitive lipase (HSL) is the rate-limiting enzyme in adipose tissue lipolysis. The aim of this experimental trial was to study the effects of a beta3-adrenergic agonist (Trecadrine) on plasma fatty acids, adipocyte HSL activity, and gene expression in control and cafeteria-induced obese animals. RESEARCH METHODS AND PROCEDURES: Control and cafeteria-fed rats were treated with a placebo or Trecadrine during 35 days. Plasma fatty acids were measured by an enzymatic method, whereas HSL activity was assessed by using labeled triolein as substrate. Finally, HSL gene expression from white adipose tissue (WAT) was determined using a reverse transcription-polymerase chain reaction method. RESULTS: Trecadrine administration reduced plasma fatty acids and HSL mRNA levels in abdominal WAT, whereas HSL activity was significantly higher in the Trecadrine-treated obese rats than in the obese nontreated rats. Also, abdominal WAT HSL activity significantly increased, whereas WAT HSL gene expression fell in control rats treated with beta3-adrenergic agonist as compared with control untreated animals. DISCUSSION: In situations of fat accumulation (high-fat feeding) or lipid mobilization (beta3-adrenergic stimulation), changes in HSL activity and HSL gene expression seem to follow a trend related to plasma fatty acids levels, as indicated by the positive correlation (r = 0.39, p < 0.05) between HSL mRNA levels and plasma fatty acids, and the negative correlation (r = -0.38, p < 0.05) between plasma fatty acids and HSL activity. Furthermore, a highly negative correlation (r = -0.59, p < 0.001) between HSL activity and HSL mRNA expression was found, in which plasma-free fatty acids are apparently involved. PMID- 10832770 TI - Prevalence of sleep apnea and electrocardiographic disturbances in morbidly obese patients. AB - OBJECTIVE: To determine the prevalence of sleep apnea in morbidly obese patients and its relationship with cardiac arrhythmias. RESEARCH METHODS AND PROCEDURES: Fifty-two consecutive morbidly obese (body mass index > or = 40 kg/m2) outpatients from the Obesity Clinic of the National Institute of Nutrition Salvador Zubiran underwent two nights of polysomnography with standard laboratory techniques. Electrocardiographic polysomnography signals (Lead II) were evaluated by two experienced cardiologists, and sleep complaints were measured with a standard sleep questionnaire (Sleep Disorders Questionnaire). In order to make comparisons between groups with different severities of sleep-disordered breathing, we classified the patients in four groups using the apnea-hypopnea index (AHI): Group 1, AHI 5 < 15 (n = 10); Group 2, AHI 15 < 30 (n = 10); Group 3, AHI 30 < 65 (n = 14); Group 4, AHI > or = 65 (n = 17). RESULTS: A wide range of sleep-disordered breathing, ranging from AHI of 2.5 to 128.9 was found. Ninety eight percent of the sample (n = 51) had an AHI > or = 5 (mean = 51 +/- 37), and 33% had severe sleep apnea with AHI > or = 65 with a mean nocturnal desaturation time of <65% over 135 minutes. Electrocardiographic abnormalities were present in 31% of the patients. Cardiac rhythm alterations showed an association with the level of sleep-disordered breathing and oxygen desaturation. DISCUSSION: We conclude that there is a high prevalence of sleep apnea in morbidly obese patients and that the risk for cardiac arrhythmias increases in this population in the presence of a severe sleep apnea (AHI > or = 65) with severe oxygen desaturation (SaO2 < or = 65%). PMID- 10832771 TI - Overweight, obesity, and blood pressure: the effects of modest weight reduction. AB - Several large epidemiological studies have shown an association between body mass index and blood pressure in normal weight and overweight patients. Weight gain in adult life especially seems to be an important risk factor for the development of hypertension. Weight loss has been recommended for the obese hypertensive patient and has been shown to be the most effective nonpharmacological treatment approach. However, long-term results of weight loss programs are disappointing with people often regaining most of the weight initially lost. In recent years, a modest weight loss, defined as a weight loss of 5% to 10% of baseline weight, has received increasing attention as a new treatment strategy for overweight and obese patients. A more gradual and moderate weight loss is more likely to be maintained over a longer period of time. Several studies have confirmed the blood pressure-lowering effect of a modest weight loss in both hypertensive and nonhypertensive patients. A modest weight loss can normalize blood pressure levels even without reaching ideal weight. In patients taking antihypertensive medication, a modest weight loss has been shown to lower or even discontinue the need for antihypertensive medication. In patients with high normal blood pressure, a modest weight loss can prevent the onset of frank hypertension. The blood pressure-lowering effect of weight loss is most likely a result of an improvement in insulin sensitivity and a decrease in sympathetic nervous system activity and occurs independent of salt restriction. In conclusion, a modest weight loss that can be maintained over a longer period of time is a valuable treatment goal in hypertensive patients. PMID- 10832772 TI - Secretin and autism: a two-part clinical investigation. AB - Recent anecdotal reports have touted the gastrointestinal (GI) hormone secretin as a treatment modality for autism, though there is little clinical evidence or literature to support its viability. We undertook a two-part clinical trial to investigate these claims. Fifty-six patients (49 boys, 7 girls, mean age = 6.4 years, SD = 2.7) enrolled in an open-label trial of secretin, during which they received one injection of the hormone (2 IU/kg). All subjects were evaluated by their parents at baseline and follow-up visits (3-6 weeks later, M = 3.7, SD = 1.4 weeks) with Childhood Autism Rating Scales (CARS). Thirty-four patients were labeled with Pervasive Developmental Disorder Not Otherwise Specified, and 22 met diagnostic criteria for Autistic Disorder. Forty-five patients were concurrently on other drug treatments. At follow-up, some reported minimal but potentially significant improvements including changes in GI symptoms, expressive and/or receptive language function, and improved awareness and social interactions. No adverse effects were reported or observed. Subsequently, 17 of the most responsive patients from Study 1 began a double-blind trial that also included 8 newly enrolled patients. Patients in this second study were alternatively entered into one of two groups and received injections of secretin or placebo with crossover at 4 weeks. Patients from Study 1 entered into Study 2 at an average of 6.5 (SD = 0.8) weeks after beginning Study 1. Results of both inquiries indicate that although treatment with secretin was reported to cause transient changes in speech and behavior in some children, overall it produced few clinically meaningful changes when compared to children given placebo injections. PMID- 10832773 TI - Comments on "Secretin and autism: a two-part clinical investigation" by M.G. Chez et al. PMID- 10832775 TI - Developmental regression in autism: maternal perception. AB - Developmental regression among children with autism is a common phenomenon of unknown origin. The purpose of this study was to identify the differences between children with autism who reportedly regressed with those who did not regress. A representative group of 39 mothers were interviewed (40 children--1 pair of twin girls) about familial, pregnancy, perinatal, as well as medical history and developmental milestones. The study focused on mothers' perceptions of developmental regression. Nineteen children (47.5%) regressed in verbal and nonverbal communication and social but not in motor abilities. Mean age of regression was 24 months, with 11 children who regressed before and 8 after this age. No significant differences were reported by mothers of children who did or did not regress. More mothers of children who regressed, than those of children who did not, expressed guilt feelings regarding the development of autism, and almost all of them had an "explanation" for the possible mechanisms that might have influenced their children's developmental course. In conclusion, developmental regression in our population appears to be a typical event in the natural course of autism. There is little difference between those children who regressed and those who did not regress in maternal perceptions and reports of development, family, and medical history. PMID- 10832774 TI - Assessment in multisite randomized clinical trials of patients with autistic disorder: the Autism RUPP Network. Research Units on Pediatric Psychopharmacology. AB - Assessment of autistic disorder (autism) symptoms, primary and secondary, poses more challenging problems than ordinarily found in multisite randomized clinical trial (RCT) assessments. For example, subjects may be uncommunicative and extremely heterogeneous in problem presentation, and current pharmacological treatments are not likely to alter most core features of autism. The Autism Research Units on Pediatric Psychopharmacology (RUPP Autism Network) resolved some of these problems during the design of a risperidone RCT in children/adolescents. The inappropriateness of the usual anchors for a Clinical Global Impression of Severity (CGI-S) was resolved by defining uncomplicated autism without secondary symptoms as a CGI-S of 3, mildly ill. The communication problems, compromising use of the patient as an informant, were addressed by several strategies, including careful questioning of care providers, rating scales, laboratory tests, and physical exams. The broad subject heterogeneity requires outcome measures sensitive to individual change over a wide spectrum of treatment response and side effects. The problems of neuropsychologically testing nonverbal, lower functioning, sometimes noncompliant subjects requires careful instrument selection/adaptation and flexible administration techniques. The problems of assessing low-end IQs, neglected by most standardized test developers, was resolved by an algorithm of test hierarchy. Scarcity of other autism-adapted cognitive and neuropsychological tests and lack of standardization required development of a new, specially adapted battery. Reliability on the Autism Diagnostic Interview (currently the most valid diagnostic instrument) and other clinician instruments required extensive cross-site training (in-person, videotape, and teleconference sessions). Definition of a treatment responder required focus on individually relevant target symptoms, synthesis of possible modest improvements in many domains, and acceptance of attainable though imperfect goals. The assessment strategy developed is implemented in a RCT of risperidone (McDougle et al., 2000) for which the design and other methodological challenges are described elsewhere (Scahill et al., 2000). Some of these problems and solutions are partially shared with RCTs of other treatments and other disorders. PMID- 10832776 TI - High functioning autism and Childhood Disintegrative Disorder in half brothers. AB - Childhood Disintegrative Disorder (CDD) is grouped with autism as a subtype of Pervasive Developmental Disorder (PDD) in ICD-10 and DSM-IV. This is the first report of autism and CDD cosegregating within a sibship. J. P. and M. P. are half brothers with the same mother. J. P. is an 18-year-old with impairments in communication, social reciprocity, and stereotypies and was diagnosed with autism. M. P. is a 7-year-old who developed normally to 2 years 4 months. He then underwent a profound regression, becoming nonverbal and socially withdrawn, and lost adaptive skills. Investigations did not reveal any neurodegenerative process. M. P. was diagnosed with CDD. The rarity of the two conditions suggests a shared transmissible mechanism. The implications for autism/PDD genetic studies are discussed. PMID- 10832777 TI - Parental report of sleep problems in children with autism. AB - This research evaluated parent reports of sleep behaviors of four groups of children: those with Autism or Pervasive Developmental Disorders, those with General Mental Retardation alone, those attending Special Education classes (with no MR diagnosis), and a control group of similar aged children without a developmental diagnosis. Diagnostic classification and demographic information were determined through parent report, report of classroom registration, and the Gilliam Autism Rating Scale (Gilliam, 1995). To evaluate sleeping behavior the study used a 28-item, five-factor scale (Behavioral Evaluation of Disorders of Sleep/BEDS; Schreck, 1997/1998) constructed from the diagnostic criteria for childhood sleep disorders found in the International Classification of Sleep Disorders: Diagnostic and Coding Manual (ICSD, American Sleep Disorders Association, 1990). Findings suggest that reports of parents with children with autistic characteristics exhibit expected quantities of sleep, but parent perception of their sleep difficulties and sleep quality is different for children with autism than for children in all other study groups. PMID- 10832778 TI - Age and IQ at intake as predictors of placement for young children with autism: a four- to six-year follow-up. AB - The predictive power of age and IQ at time of admission to an intensive treatment program using applied behavior analysis were examined in a 4- to 6-year follow-up of educational placement. Twenty-seven children with autistic disorder who were between the ages of 31 and 65 months and had IQs on the Stanford Binet between 35 and 109 at time of admission to the Douglass Developmental Disabilities Center were followed up 4 to 6 years after they left the preschool. The results showed that having a higher IQ at intake (M = 78) and being of younger age (M = 42 months) were both predictive of being in a regular education class after discharge, whereas having a lower IQ (M = 46) and being older at intake (M = 54 months) were closely related to placement in a special education classroom. The results are interpreted as pointing to the need for very early intervention for children with Autistic Disorder. It is also emphasized that older children and those with lower IQs in the present study showed measurable gains in IQ from treatment. The data should not be taken to suggest that children older than 4 years of age do not merit high quality treatment. PMID- 10832779 TI - The development of siblings' understanding of autism spectrum disorders. AB - While professionals commonly advocate sharing information about autism spectrum disorders with siblings, no guidelines currently exist that describe what types of information might be relevant for siblings at different ages. To address this issue, the interviewing method described by Bibace and Walsh (1979, 1980), which measures cognitive sophistication in thinking about illness, was adapted to examine perspectives on autism spectrum disorders. Sixty-three siblings of individuals with autism or related disorders were interviewed using this measure. Parents were given the same interview as their child, and asked to predict their child's responses. Children's reasoning became more mature with age, but developed at a delayed rate compared to norms for illness concepts. Although accurate in estimating their child's understanding of the definition and cause of their sibling's diagnosis, parents tended to overestimate their child's understanding of the disorder's impact. PMID- 10832780 TI - Brief report: Recognition of autism spectrum disorder before one year of age: a retrospective study based on home videotapes. PMID- 10832781 TI - Brief report: Interrater reliability of clinical diagnosis and DSM-IV criteria for autistic disorder: results of the DSM-IV autism field trial. PMID- 10832782 TI - Brief report: Comparison of sensory-motor and cognitive function between autism and Asperger syndrome in preschool children. PMID- 10832783 TI - Is the cognitive style of the persons with the Asperger's syndrome also a "mathematical style"? PMID- 10832784 TI - What is Childhood Disintegrative Disorder, how is it different from autism, and what is believed to be its cause? PMID- 10832785 TI - The effects of neurotoxic hippocampal lesions on two effects of context after fear extinction. AB - Three conditioned suppression experiments with rats examined the role of the hippocampus in 2 effects of context after extinction. Reinstatement is the context-specific recovery of fear to an extinguished conditioned stimulus (CS) that occurs following independent presentations of the unconditioned stimulus (US), after extinction. Renewal is the recovery of fear when the CS is presented in the context in which it was conditioned, after extinction in a different context. Results indicated that neurotoxic lesions of the hippocampus, performed before conditioning, abolished reinstatement, which depends on context-US associations, but not renewal, which does not. This dissociation is not the result of differences in the recentness of context learning that ordinarily governs the 2 effects. The results suggest that the hippocampus is necessary for some, but not all, types of contextual learning. PMID- 10832786 TI - Arousal-related associative response characteristics of dorsal lateral geniculate nucleus neurons during acoustic Pavlovian fear conditioning. AB - This research determined whether fear-conditioned, acoustic stimuli induce thalamic arousal reflected in associative responses in dorsal lateral geniculate nucleus (dLGN) neurons. Rabbits received a Pavlovian discriminative fear conditioning procedure in which one tone conditioned stimulus (CS +) was always paired with an aversive unconditioned stimulus (US) and another tone (CS-) was never paired with the US. Responses of single dLGN neurons to random CS+ and CS- presentations were then recorded. Nine of 15 recorded neurons demonstrated significantly greater firing during the CS+ versus the CS-. Their spontaneous activity demonstrated tonic firing during increased neocortical arousal and burst firing during decreased neocortical arousal. The results demonstrate that dLGN neurons show associative responses to fear-conditioned, acoustic stimuli and present a model for investigating the neural circuits by which such stimuli affect sensory processing at the thalamic level. PMID- 10832787 TI - Learning- and cerebellum-dependent neuronal activity in the lateral pontine nucleus. AB - The effects of inactivation of cerebellar deep nuclei and the lateral pontine nucleus on classical eyeblink conditioning with tone or lateral reticular nucleus (LRN) stimulation as conditioned stimuli (CSs) were examined. Inactivation of cerebellar deep nuclei abolished eyeblink conditioned responses (CRs) when the CS was either a tone or LRN stimulation. Inactivation of the lateral pontine nucleus prevented only the acquisition and retention of tone-evoked eyeblink CRs. Multiple-unit recording demonstrated that when LRN stimulation was used as the CS, inactivation of the interpositus nucleus abolished learning-related neuronal activity in the lateral pontine nucleus, whereas inactivation of pontine nucleus had little effect on similar activity in the interpositus nucleus. Thus, the learning-induced neuronal activity in the lateral pontine nucleus was most likely driven by the cerebellar interpositus nucleus. PMID- 10832788 TI - Peripheral and intraamygdalar administration of the dopamine D1 receptor antagonist SCH 23390 blocks fear-potentiated startle but not shock reactivity or the shock sensitization of acoustic startle. AB - Central dopamine (DA) activity is thought to play a role in fear motivation. The aim of the present study was to assess the involvement of DA D1 receptors in emotional learning. The authors report that peripheral and intraamygdalar administration of the specific D1 receptor antagonist SCH 23390 blocked the acquisition of fear-potentiated startle. Analysis of shock reactivity during footshock administration revealed that the learning impairment could not be explained by a diminution in the aversive properties of the unconditioned stimulus. Additionally, systemic and intraamygdalar injection of SCH 23390 did not alter fear expression as measured with the shock sensitization of acoustic startle. The potential contribution of mesoamygdaloid DA to the acquisition and retrieval of conditioned fear responses is discussed. PMID- 10832789 TI - The place preference task: a new tool for studying the relation between behavior and place cell activity in rats. AB - This study describes a task that combines random searching with goal directed navigation. The testing was conducted on a circular elevated open field (80 cm in diameter), with an unmarked target area (20 cm in diameter) in the center of 1 of the 4 quadrants. Whenever the rat entered the target area, the computerized tracking system released a pellet to a random point on the open field. Rats were able to learn the task under light and in total darkness, and on a stable or a rotating arena. Visual information was important in light, but idiothetic information became crucial in darkness. Learning of a new position was quicker under light than in total darkness on a rotating arena. The place preference task should make it possible to study place cells (PCs) when the rats use an allothetic (room frame) or idiothetic (arena frame) representation of space and to compare the behavioral response with the PCs' activity. PMID- 10832790 TI - Pedunculopontine tegmental nucleus lesions impair stimulus--reward learning in autoshaping and conditioned reinforcement paradigms. AB - The role of the pedunculopontine tegmental nucleus (PPTg) in stimulus-reward learning was assessed by testing the effects of PPTg lesions on performance in visual autoshaping and conditioned reinforcement (CRf) paradigms. Rats with PPTg lesions were unable to learn an association between a conditioned stimulus (CS) and a primary reward in either paradigm. In the autoshaping experiment, PPTg lesioned rats approached the CS+ and CS- with equal frequency, and the latencies to respond to the two stimuli did not differ. PPTg lesions also disrupted discriminated approaches to an appetitive CS in the CRf paradigm and completely abolished the acquisition of responding with CRf. These data are discussed in the context of a possible cognitive function of the PPTg, particularly in terms of lesion-induced disruptions of attentional processes that are mediated by the thalamus. PMID- 10832791 TI - Effects of US devaluation on win-stay and win-shift radial maze performance in rats. AB - Previous studies have shown double dissociations between win-stay and win-shift radial maze learning in terms of their underlying neural substrates. To examine the content of the associations formed in the two tasks, the authors devalued the food unconditioned stimulus (US) by taste aversion to differentiate stimulus stimulus(CS-US) and stimulus-response (CS-CR) learning. US devaluation was performed in rats that were over- or undertrained on the win-stay task. Devaluation substantially reduced food consumption on the maze but failed to disrupt choice accuracy, regardless of the amount of training. Devaluation did not affect latency in overtrained rats but did increase latency in undertrained rats. In the win-shift task, devaluation caused rats to reject the reinforcer, yet they continued to accurately win-shift, but with significantly longer latencies (Experiment 3). The results suggest that an S-R association may mediate performance after extended win-stay training. In contrast, a US representation appears to be recalled during early win-stay and win-shift performance. PMID- 10832792 TI - Glucose attenuation of memory impairments. AB - The performance of pigeons in a short-term memory procedure (delayed matching-to sample) was studied over a range of retention intervals from 0.2 s to 24.0 s. The authors examined the ability of 3 dose levels of glucose (0, 50, and 100 mg/kg) to alleviate memory impairments produced by administration of scopolamine (0.03 mg/kg), by a reduction in the sample-response requirement and by interpolating retroactive interference in the retention interval (houselight illumination). Glucose administration attenuated the deficit produced by scopolamine and by the reduced sample-response requirement, by reversing the decrement in accuracy at 0 delay. Glucose did not, however, reverse the increase in rate of forgetting generated by retroactive interference. The results suggest that the mode of action by which glucose is able to attenuate drug-induced and behavioral impairments in memory may be through an effect on attentional or encoding processes. PMID- 10832793 TI - Anticipation of rewarding electrical brain stimulation evokes ultrasonic vocalization in rats. AB - Adult rats emit increased rates of 50-kHz ultrasonic vocalizations (USVs) before receiving social and pharmacological rewards. This study sought to determine whether anticipation of rewarding electrical stimulation of the brain (ESB) would also elicit these vocalizations. In Experiments 1 and 2, rats showed increased 50 kHz USVs before receiving experimenter-delivered ventral tegmental area (VTA) and lateral hypothalamic (LH) ESB on a fixed time 20-s schedule. In Experiments 3 and 4, rats increased their rate of 50-kHz USVs in response to cues that predicted the opportunity to self-stimulate the VTA or LH. Interestingly, unexpected termination of either type of ESB evoked 20-kHz, rather than 50-kHz, USVs. In Experiment 5, a cue that predicted daily 1-hr feeding sessions increased 50-kHz USVs, whereas a cue that predicted footshock decreased 50-kHz USVs. These effects could not be explained simply by changes in locomotor activity or general arousal. Together, these findings support the hypothesis that short 50-kHz USVs may selectively index a state of reward anticipation in rats. PMID- 10832794 TI - Spontaneous motor activity in fetal and infant rats is organized into discrete multilimb bouts. AB - Spontaneous motor activity (SMA) is a ubiquitous feature of fetal and infant behavior. Although SMA appears random, successive limb movements often occur in bouts. Bout organization was evident at all ages in fetal (embryonic day [E] 17 21) and infant (postnatal day [P] 1-9) rats, with nearly all bouts comprising 1-4 movements of different limbs. A computational model of SMA, including spontaneous activity of spinal motor neurons, intrasegmental and intersegmental interactions, recurrent inhibition, and descending influences, produced bouts with the same structure as that observed in perinatal rats. Consistent with the model, bouts were not eliminated on E20 after cervical spinal transection, suggesting that the brain is not necessary to produce bout organization. These investigations provide a foundation for understanding the contributions of SMA to neuromuscular and motor development. PMID- 10832795 TI - Using c-Fos immunocytochemistry to identify forebrain regions that may inhibit maternal behavior in rats. AB - Evidence indicates there is a neural system that inhibits maternal behavior in virgin rats. It has been suggested that pregnancy hormones promote the onset of maternal behavior by reducing the behavioral influence of this system. The authors used c-Fos immunocytochemistry to identify brain regions more activated by pup exposure in nonmaternal rats than in maternal rats. Previous experiments indicated that some of these regions, such as the posterodorsal medial amygdala and several medial hypothalamic sites, inhibit maternal behavior. For others, such as the ventral lateral septum, dorsal premammillary nucleus, and principal bed nucleus of the stria terminalis, this is the first indication that they could also inhibit maternal responding. These regions have previously been implicated in promoting defensive behaviors, consistent with the finding that nonmaternal rats actively avoid pups. These findings suggest the existence of a neural circuit through which pup exposure could promote defensive responses in virgin rats, and how pregnancy hormones could reduce such activity to stimulate maternal behavior. PMID- 10832796 TI - The suppressive effects of sucrose and cocaine, but not lithium chloride, are greater in Lewis than in Fischer rats: evidence for the reward comparison hypothesis. AB - Rats suppress intake of a saccharin conditioned stimulus (CS) when it is paired with an aversive unconditioned stimulus (US), an appetitive US, or a drug of abuse such as morphine or cocaine. It is unclear, however, whether the reduction in intake induced by these drugs is mediated by their aversive or their rewarding properties. The present set of experiments addressed this question by comparing the suppressive effects of a known aversive US (LiCl), a known reinforcing US (sucrose), and a drug of abuse (cocaine) in two strains of rats (i.e., Lewis and Fischer 344 rats) that differ in their preference for rewarding stimuli. The results show that, although both strains readily acquired a LiCl-induced conditioned taste aversion (CTA), the suppressive effects of sucrose and cocaine were robust in the drug-preferring Lewis rats and absent in the Fischer rats. These data argue against a CTA account and in favor of the reward comparison hypothesis. PMID- 10832797 TI - Strain differences in the isolation-induced effects on prepulse inhibition of the acoustic startle response and on locomotor activity. AB - The authors investigated the effects of isolation rearing on acoustic startle response, prepulse inhibition (PPI), its modification by apomorphine, and locomotor activity in 3 rat strains: Wistar (WS), Sprague-Dawley (SD), and Lister hooded (LH). SD and LH, but not WS, showed isolation-induced PPI deficits. In 2 consecutive PPI tests, only SD isolates showed significant PPI deficits. An isolation rearing effect in LH was significant only in the 1st PPI test. Apomorphine dose-dependently (0.0-0.5 mg/kg) disrupted PPI, but sensitivity to the drug differed, with WS and SD rats being more sensitive to lower doses (0.01 0.05 mg/kg) than LH rats (0.5 mg/kg). Isolates, irrespective of strain, did not differ from grouped rats in their response to the apomorphine challenge. Only WS and LH isolates demonstrated significantly increased locomotor activity. Strain differences in the different parameters measured did not predict isolation induced effects on PPI. PMID- 10832798 TI - Prepulse startle deficit in the Brown Norway rat: a potential genetic model. AB - Prepulse inhibition (PPI), an operational measure of sensorimotor gating, is deficient in schizophrenia patients. PPI was compared among 4 strains of rats: Sprague-Dawley, Spontaneously Hypertensive, Wistar Kyoto (WKY), and Brown Norway (BN). PPI was dramatically lower in BN versus the other strains, especially WKY, for both acoustic and airpuff startle stimuli, whereas startle amplitude was similar between BN and WKY. Female BN also had lower PPI than did female WKY. Response to increasing prepulse intensities showed a right shift in the BN relative to the WKY. Visual prepulses also showed deficiencies in BN versus WKY. The absence of background noise did not negate strain differences. Auditory brainstem response to clicks and tone pips revealed no differences in auditory threshold between the 2 strains. These results are the first to demonstrate that BN have impaired sensorimotor gating compared with WKY, without impaired acoustic acuity. PMID- 10832799 TI - Different inhibition of conditioned avoidance response by clozapine and DA D1 and D2 antagonists in male mice. AB - The effects of clozapine (2.5 and 5 mg/kg), SCH 23390 (0.05 and 0.1 mg/kg), and raclopride (0.1 and 0.5 mg/kg) on the acquisition and performance of a conditioned avoidance response (CAR) were studied in BALB/C mice. The high dose of clozapine decreased avoidances and crossings in acquisition and performance. SCH 23390 had no effect on acquisition, whereas a decrease of avoidances and crossings was produced by the high dose in performance. The high dose of raclopride decreased avoidances and crossings in acquisition but had no effects on performance. The results suggest that the mechanisms by which these drugs affect avoidance are not the same. This difference may reflect an action on different subtypes of DA receptors that produces different effects on motor behavior. It seems that in all cases where CAR is impaired, locomotor activity is also suppressed; therefore, a parsimonious interpretation is that the CAR procedure is sensitive to motor effects. PMID- 10832800 TI - Sensitivity to ethanol-induced motor incoordination in 5-HT(1B) receptor null mutant mice is task-dependent: implications for behavioral assessment of genetically altered mice. AB - Neuromuscular impairment by ethanol likely involves complex effects on balance, gait, muscle strength, and other features of motor coordination. The present experiments showed that relative sensitivity to ethanol-induced motor impairment in serotonin 1B (5-HT(1B)) null mutant and control mice was task dependent. We found that ethanol-treated null mutant mice made fewer missteps on a balance beam than did ethanol-treated wild-type mice, and confirmed a previous finding of their lesser ethanol sensitivity in the grid test. The genotypes did not differ in ethanol sensitivity as measured by the screen test, static dowel, fixed-speed rotarod, accelerating rotarod, grip strength, or loss of righting reflex tests. These experiments suggest that within a behavioral domain, alternative tests of function are not equivalent, so multiple assessment tools should be used to avoid misinterpretation of gene function. PMID- 10832801 TI - Functional recovery of respiratory behavior during axonal regeneration in snails (Lymnaea stagnalis) is experience dependent. AB - This study investigated the role of experience in recovery of pulmonary respiration during axonal regeneration in Lymnaea stagnalis. Pulmonary respiration occurs when snails break the water surface and open the lung orifice, the pneumostome. It was shown that axotomy of all the axons innervating the pneumostome and surrounding area prevents the occurrence of lung respiration in 69% of snails. In the remaining 31%, lung respiration persisted, indicating that peripheral components alone are capable of initiating pneumostome openings and closures. Five weeks postsurgery, all snails with previous nerve crushes showed opening of the pneumostome with normal latency after breaking the water surface. However, prevention of pulmonary respiration during the recovery period dramatically changed the recovered behavior. Thus, experience in pulmonary respiration during axonal regeneration plays a role in the recovery of this behavior. PMID- 10832802 TI - Naloxone-induced opiate withdrawal produces long-lasting and context-independent changes in aggressive and social behaviors of postdependent male mice. AB - The purpose of this study was to determine whether an environment associated with naloxone-induced morphine withdrawal affects aggressive or social behaviors in postdependent mice. Morphine-dependent or saline-treated mice received 3 naloxone injections in 1 of 2 different environments (A or B); 15 days afterward, when the mice were completely drug free, an aggression test was carried out in Environment A. All the mice suffering morphine withdrawal showed a significant increase in aggression, irrespective of the environment in which the withdrawal took place. In these conditions, the impact of morphine dependence and the 3 induced withdrawals was so profound that the environment could not be discriminative. In addition, modifications in the behavioral profile of postdependent mice that suffered only spontaneous withdrawal were long-lasting, with the mice carrying out more attacks during social investigation without presenting threat postures. PMID- 10832803 TI - Behavioral recovery after transplantation into a rat model of Huntington's disease: dependence on anatomical connectivity and extensive postoperative training. AB - Rats were trained to perform a conditioned stimulus-response task known to be sensitive to striatal damage, after which they received unilateral excitotoxic striatal lesions. The subsequent implantation of graft tissue into the lesioned striatum was either immediate (9 days) or substantially delayed (70 days). When retested 14 weeks later, all graft and lesion rats were equally impaired initially and biased their responding toward the ipsilateral side. Graft associated recovery was evident with repeated postoperative testing, but only in rats that had received transplants 9 days postlesion. It is suggested that this training-dependent, graft-associated recovery is mediated specifically by the restored host-graft connections. PMID- 10832804 TI - Behavioral taste responses of developmentally NaCl-restricted rats to various concentrations of NaCl. AB - The behavioral taste responses of developmentally NaCl-restricted rats were examined with a brief-exposure taste test. Neurophysiological and morphological alterations have been reported in rats whose dietary sodium is restricted during pre- and postnatal development, yet there exists little data discerning their behavioral response to tastants. Control and developmentally NaCl-restricted rats were maintained on a low-NaCl diet and trained to lick from individually presented sipper tubes. Each subject received 4 days of testing on various NaCl concentrations. Results indicate that developmentally NaCl-restricted rats have dramatically increased lick rates to NaCl solutions. These responses are likely due to some combination of factors including (a) numbers and type of active chorda tympani fibers, (b) compensatory responses to NaCl-solutions from other nerves of the oral cavity, and (c) increased sensitivity of central taste systems to NaCl. PMID- 10832805 TI - Rats alternate on a dry-land but not swimming-pool (Morris task) place task: implications for spatial processing. AB - Groups of rats were rewarded with food for traveling from a start point to 2 different locations while their alternations in choice between those locations on 2 daily trials were recorded. In one experimental condition, the rats swam and received food once they climbed upon a platform that was hidden just below the surface of the water at the food location. In the other condition, the rats walked to reach the food. It was found that the rats did not alternate their choices between target locations when swimming but that they did alternate target choices when walking. Even experience in alternating when walking did not produce reliable alternation when swimming. It is proposed that rats treat escape (swimming) and search (walking) tasks in different ways, and this difference is discussed with respect to the possibility that different central processes may be used in the task solutions. PMID- 10832806 TI - Gene-specific differences in the circadian variation of ventricular repolarization in the long QT syndrome: a key to sudden death during sleep? AB - BACKGROUND: In the long QT syndrome (LQTS) most life-threatening cardiac events occur in association with physical or emotional stress. However, a minority of patients dies suddenly during sleep; intriguingly, these sleep-related sudden deaths tend to cluster in families. The mechanism(s) underlying this phenomenon and the reason why it occurs in few selected families are unknown. Recently, some of the LQTS genes have been identified leading to three main subgroups (LQT1, LQT2, LQT3) associated respectively with mutations affecting the following ionic currents involved in the control of ventricular repolarization: I(Ks), I(Kr), I(Na). We have recently observed that cardiac events nighttime are rare in LQT1 and frequent in LQT3 patients. METHODS: We studied 26 LQTS patients all genotyped (11 LQT1, 9 LQT2, 6 LQT3) and 26 healthy controls matched by age and gender. Using a specific software, 24-hour ambulatory ECG recordings were performed and the QT interval was measured in order to allow comparison between QTc nighttime and daytime. RESULTS: The main finding is that while LQT1 patients show a trend for modest QTc shortening and LQT2 patients a trend for modest lengthening nighttime versus daytime, LQT3 patients show clear lengthening of the QTc nighttime. These changes are not explained by heart rate changes or by the use of beta-blockers. CONCLUSIONS: The marked tendency for further QT prolongation nighttime, which clearly increases arrhythmic risk, present among LQT3 patients and absent among LQT1 patients, provides an explanation for the gene-specific higher risk for sudden death during sleep for LQT3 compared to LQT1 patients. PMID- 10832807 TI - Sleep as an autonomic stress test for the heart. PMID- 10832808 TI - Influence of hydrophilic and lipophilic beta-blockers on heart rate, ventricular repolarization and their interrelationship in normal subjects. AB - BACKGROUND: It has been hypothesized that hydrophilic and lipophilic beta blockers have different antiarrhythmic properties because only the latter seem to reduce the rate of sudden death in post-myocardial infarction patients as well as animal models which seem to be independent of their effect on autonomic nervous system modulation. The aim of this study was to evaluate the different effects of a hydrophilic (nadolol) and lipophilic (metoprolol) beta-blocker on ventricular repolarization in normal subjects. METHODS: Seventeen normal subjects entered this randomized, single-blind cross-over study designed to compare the effects of nadolol (80 mg/day) and slow-release metoprolol (200 mg/day) on dynamic ventricular repolarization. The RR intervals, the QT evaluated at the apex (QT apex) and at the end (QT end) of the T wave before and after correction for heart rate, the standard deviation of QT apex and QT end, and the slope of the QT/RR linear relationship (QTa-slope and QTe-slope) were studied using the ELATEC system (ELA Medical, Mountrouge, France), and an evaluation was made of their reproducibility and the effects of each beta-blocker. RESULTS: The most reproducible parameters were QT apex, corrected QT apex and the QTe-slope. Nadolol was associated with a greater adrenergic blockade than metoprolol (lengthening of RR interval +25 +/- 7 and +17 +/- 8% respectively, p = 0.0003) and a lower effect on ventricular repolarization (reduction of corrected QT apex 0.6 +/- 3 and -2.5 +/- 2.1% respectively, p < 0.01; reduction of QTe-slope -5 +/- 16 and -15 +/- 15% respectively, p = 0.03). CONCLUSIONS: At the dosages used in the study, metoprolol showed lower adrenergic blockade but greater effect on ventricular repolarization than nadolol. PMID- 10832809 TI - Early administration of verapamil after thrombolysis in acute anterior myocardial infarction. Effect on left ventricular remodeling and clinical outcome. VAMI Study Group. Verapamil Acute Myocardial Infarction. AB - BACKGROUND: The administration of verapamil during the reperfusion phase of acute myocardial infarction can reduce the extent and severity of microvessel damage and limit myocardial dysfunction. We aimed at investigating the effect of early verapamil administration on left ventricular remodeling and the clinical evolution after myocardial infarction. METHODS: Eighty-eight patients with first acute anterior myocardial infarction thrombolysed < 4 hours from symptom onset were enrolled in a multicenter, randomized, double-blind, controlled study of verapamil administration (5 mg i.v. + 2 microg/kg/min over 24 hours). Echocardiographic end-diastolic (EDV) and end-systolic (ESV) left ventricular volumes were assessed by biplane Simpson's rule. RESULTS: At 90 days, EDV in the verapamil and placebo groups was respectively 88.9 +/- 27.8 and 95.8 +/- 30.7 ml (p = 0.11), ESV was 52.6 +/- 22.7 and 57.7 +/- 25.4 ml (p = 0.18). There was no change over time in the verapamil group (day 3 vs day 90: EDV 85.0 +/- 17.7 vs 88.9 +/- 27.8 ml, p = NS; ESV 48.7 +/- 14.1 vs 52.6 +/- 22.7 ml, p = NS) while left ventricular volume increased in the placebo group (day 3 vs day 90: EDV 87.6 +/- 21.1 vs 95.8 +/- 30.7 ml, p = 0.03; ESV 52.0 +/- 16.9 vs 57.7 +/- 25.4 ml, p = 0.08). NYHA functional classes were differently distributed at 30 and 90 days (chi2 = 0.009 and 0.07), with a lower prevalence of classes II and III in the verapamil group (p = 0.03). CONCLUSIONS: The early intravenous administration of verapamil in thrombolysed patients can reduce left ventricular remodeling and NYHA functional class after acute anterior myocardial infarction. PMID- 10832810 TI - Increasing plasma homocysteine during follow-up in heart transplant recipients: effects of folate and renal function. AB - BACKGROUND: Hyperhomocysteinemia is a common finding in heart transplant recipients and may represent a risk factor for graft failure. However, the time course, determinants and effects of medical therapy on total homocysteine plasma levels after heart transplantation remain undetermined. The aim of this study was to prospectively analyze 1) the time-course of total homocysteine in heart transplant recipients; 2) the effects of folate supplements and cyclosporine A on total homocysteine; 3) the relation among renal function, serum vitamin levels, and total homocysteine. METHODS: Fifty-two heart transplant recipients consecutively evaluated for routine follow-up during 1998 were included in the study (mean age 54 +/- 12 years; 28% female). Among the 52 patients, 10 patients were treated with folate for the entire period of the study (Group F), while 26 patients never received folate (Group NF). The remaining 16 patients who did not take folate on a regular basis were excluded from subgroup analysis. Total homocysteine and creatinine plasma levels were assayed at entry into the study (time 0) and at the end of the study, 12 months later (time 12). RESULTS: Homocysteinemia increased significantly from time 0 to time 12 (p < 0.001), regardless of creatinine plasma levels (p = 0.03) and folate intake (p < 0.01). However, total homocysteine levels were lower in Group F compared to Group NF at time 0 and time 12 (p < 0.02). On multivariate analysis, time of follow-up, serum creatinine and lack of folate intake were positive independent predictors of total homocysteine. CONCLUSIONS: Homocysteinemia increased over time in heart transplant recipients, regardless of renal function and folate administration. Lower total homocysteine levels were associated with folate intake, suggesting that folate supplements may play a role in the prevention of vascular allograft disease. PMID- 10832811 TI - Cardioversion of persistent atrial flutter in non-anticoagulated patients at low risk for thromboembolism. AB - BACKGROUND: The true risk of thromboembolic events after cardioversion of atrial flutter was not addressed carefully. Nevertheless, thromboembolic events were thought to be rare and less likely to occur after cardioversion of atrial fibrillation. The aim of this study was to prospectively evaluate if the interruption of persistent typical atrial flutter could be safely performed without anticoagulation in a group of patients at low risk for thromboembolic events. METHODS: We studied 64 subjects selected among 138 consecutive patients with persistent typical atrial flutter (minimal duration 72 hours) in whom a transesophageal atrial pacing was performed in our electrophysiology laboratory from October 1994 to May 1999. Exclusion criteria included: anticoagulation therapy during the previous 4 weeks; previous history of atrial fibrillation; recent (< 1 month) myocardial infarction; history of thromboembolic events; left ventricular ejection fraction < 40%; presence of moderate or severe mitral regurgitation or stenosis; induction of sustained (> 6 hours) atrial fibrillation during transesophageal atrial pacing. Patients in whom atrial flutter persisted in spite of transesophageal atrial pacing underwent external direct current cardioversion or right atrial overdrive pacing within 24 hours. Thromboembolic events were checked for 4 weeks after the restoration of sinus rhythm. RESULTS: Sinus rhythm was restored in 54 patients by transesophageal atrial pacing, in 8 patients by electrical cardioversion, and in 2 by right atrial pacing. The mean duration of atrial flutter was 18 +/- 19 days, the mean left atrial size 41.3 +/- 6.2 mm, and the mean left ventricular ejection fraction 54.8 +/- 7.3%. During the study period no episodes of thromboembolism were recorded. CONCLUSIONS: Cardioversion of persistent typical atrial flutter in non-anticoagulated patients at low risk for thromboembolic events appears safe. PMID- 10832812 TI - Persistence of increased left ventricular mass despite optimal blood pressure control in hypertension. AB - BACKGROUND: Left ventricular hypertrophy is an adverse risk marker in essential hypertension and its regression has a favorable effect on prognosis. It is unclear whether blood pressure normalization induced by long-term therapy is able to normalize left ventricular mass completely. METHODS: In the setting of a prospective cohort study, 107 consecutive hypertensive patients who achieved blood pressure normalization (clinic blood pressure < 140/90 mmHg on > or = 3 consecutive visits) under long-term (1-10 years, average 2.9) drug treatment were individually matched with 107 healthy normotensive controls by gender, age (+/- 5 years), body mass index (+/- 3 kg/m2), and clinic systolic blood pressure (+/- 5 mmHg) in a case-control design. All subjects underwent 24-hour blood pressure monitoring and M-mode echocardiography. RESULTS: Treated hypertensive patients and normotensive controls did not differ by age, body mass index, clinic blood pressure (128/82 vs 128/81 mmHg), and 24-hour blood pressure (120/77 vs 120/76 mmHg). Left ventricular mass and relative wall thickness were greater in the hypertensive than in the normotensive group (97 +/- 24 vs 86 +/- 17 g/m2 and 0.40 +/- 0.08 vs 0.37 +/- 0.08, both p < 0.001). CONCLUSIONS: Left ventricular mass is greater in well-controlled hypertensive patients than in normotensive controls matched by age, obesity, gender, and clinic and 24-hour blood pressure. This finding is consistent with the lower than epidemiologically expected reduction in coronary heart disease risk during antihypertensive therapy and might reflect the persistent effect on left ventricular mass of hemodynamic and/or non-hemodynamic factors other than blood pressure in treated patients with essential hypertension. PMID- 10832813 TI - Vascular network changes in the retina during ageing in normal subjects: a computerized quantitative analysis. AB - BACKGROUND: Direct ophthalmoscopic examination of fundus oculi is widely used for the qualitative evaluation of target organ damage in several pathological conditions. At present, there are no reliable techniques available to quantify retinal vascular damage. The aim of this study was to develop a computerized technique for the quantitative analysis of fundus oculi. METHODS: We studied 68 non-smoking, normotensive normal subjects, with a visual acuity > 8/10. From each subject retinal images were taken using a non-mydriatic ophthalmoscope and digitized onto a personal computer. In each image the area of analysis was set to a circular selection, corresponding to 25% of the whole fundus oculi, concentric with the optic disc. From each selection arterial and venous area and mean arteriolar bifurcation angle were obtained. Measurements were taken in a blinded way by two operators and, by the same operator, twice at different times. Parameters were correlated with age and body surface area. RESULTS: Retinal arterial density (arteries 7.44 +/- 1.25%; bifurcation angle 75 +/- 16 degrees) was significantly correlated with age, but not with body surface area. Intra- and interobserver coefficient of variation resulted 2.5 and 3.2%, respectively. CONCLUSIONS: Computerized analysis of the fundus oculi with the evaluation of vascular density indexes represents a simple and reproducible technique that could be useful in identifying changes in retinal vascular network during ageing. PMID- 10832814 TI - Transient microvascular vasoconstriction: a possible cause of unstable angina. AB - We report the case of a 65-year-old woman who developed unstable angina 2 months after successful coronary angioplasty of the left anterior descending coronary artery. Coronary angiography failed to show angiographic restenosis, but intracoronary ergonovine caused ST segment elevation and her habitual chest pain in the absence of epicardial coronary spasm and important pressure changes in the distal left anterior descending coronary artery assessed by a pressure wire, thus suggesting that distal vessel constriction was responsible for unstable angina. PMID- 10832815 TI - Coronary aneurysms and stenosis detected with magnetic resonance coronary angiography in a patient with Kawasaki disease. AB - Coronary artery abnormalities in Kawasaki syndrome develop in about 15-25% of young patients, mostly as aneurysms. In the long-term evolution of coronary artery disease thrombotic occlusion of aneurysms, premature atherosclerosis, and stenosis, are observed. Magnetic resonance is an emerging modality in the angiographic assessment of coronary arteries. The authors report a case of coronary artery aneurysms and stenosis in a 26-year-old patient with Kawasaki disease detected by magnetic resonance coronary angiography and confirmed by conventional coronary angiography. PMID- 10832816 TI - Isolated noncompaction of the ventricular myocardium. A study in an adult male and literature review. AB - Noncompaction of the ventricular myocardium is a rare congenital disorder characterized by the presence of numerous prominent trabeculations and deep intertrabecular recesses which communicate with the left ventricular cavity. The disease uniformly affects the left ventricle, sometimes also affecting the right ventricle. Noncompaction of the ventricular myocardium is believed to be a disorder of endomyocardial embryogenesis. Familial occurrence has been observed. It may be accompanied by depressed ventricular function, cardiac arrhythmia and systemic embolism. Although noncompaction of the ventricular myocardium is a congenital myocardial disorder, the onset of symptoms is frequently delayed until adulthood. We describe a case of noncompaction of the ventricular myocardium in a 33-year-old male with the typical echocardiographic and cardiac magnetic resonance imaging features of this disease. PMID- 10832817 TI - Violence and the doctor's role. PMID- 10832818 TI - Calcium antagonists and secondary prevention of myocardial infarction; [comment]. PMID- 10832819 TI - Radiotherapy for early breast cancer: was a comprehensive overview of trials needed? PMID- 10832820 TI - Strengthening of the case for organised trauma-care systems. PMID- 10832821 TI - Eosinophil chemokines and their receptors: an attractive target in asthma? PMID- 10832822 TI - Assessment and management of functional or ischaemic mitral regurgitation. PMID- 10832823 TI - Violence. PMID- 10832824 TI - Association of tumour site and sex with survival benefit from adjuvant chemotherapy in colorectal cancer. AB - BACKGROUND: Adjuvant chemotherapy can improve 5-year survival in Dukes' C colorectal carcinoma. Improved selection of patients who will respond to adjuvant treatments is required. We investigated whether site of tumour origin, sex, and presence of microsatellite instability (MSI) phenotype were associated with a survival benefit from adjuvant chemotherapy. METHODS: We analysed data for 656 consecutive patients with Dukes' C colorectal carcinoma, with median follow-up of 54 months (range 7-104) and mean age 66.7 years (SD 12.9). We screened tumour samples by PCR for deletions in the BAT-26 mononucleotide repeat to establish MSI status. Details of chemotherapy and survival were obtained by review of hospital and health-department records. Adjuvant chemotherapy (fluorouracil and levamisole) was given with curative intent to 272 (42%) patients. FINDINGS: Striking survival benefits were seen for patients who had right-sided tumours and who received adjuvant chemotherapy compared with those who did not (48 vs 27% alive at end of study [95% CI 0.25-0.56], p<0.0001), for women (53 vs 33% [0.25 0.56], p<0.0001), and for patients with MSI tumours (90 vs 35% [0.01-0.53], p=0.0007). MSI-positive tumours were slightly more frequent in women than in men (10 vs 7%). Right-sided tumours were more frequently MSI positive than left-sided tumours (20 vs 1%). Men with right-sided tumours benefited from chemotherapy (37 vs 12% [0.24-0.69], p=0.0007) but men with left-sided tumours did not. INTERPRETATION: The survival benefits seen in patients treated with adjuvant chemotherapy suggest that data from previous trials of adjuvant chemotherapy should be reassessed and the predictive value of MSI status confirmed. Validation of our results will allow better selection of patients for chemotherapy. PMID- 10832825 TI - Diltiazem in acute myocardial infarction treated with thrombolytic agents: a randomised placebo-controlled trial. Incomplete Infarction Trial of European Research Collaborators Evaluating Prognosis post-Thrombolysis (INTERCEPT) AB - BACKGROUND: Diltiazem reduces non-fatal reinfarction and refractory ischaemia after non-Q-wave myocardial infarction, an acute coronary syndrome similar to the incomplete infarction that occurs after successful reperfusion. We postulated that this agent would reduce cardiac events in patients after acute myocardial infarction treated initially with thrombolytic agents-a clinical application previously unexplored with heart-rate-lowering calcium antagonists. METHODS: A prospective, randomised, double-blind, sequential trial was done in 874 patients with acute myocardial infarction, but without congestive heart failure, who first received thrombolytic agents. Patients received either 300 mg oral diltiazem once daily, or placebo, initiated within 36-96 h of infarct onset, and given for up to 6 months. The trial primary endpoint was the cumulative first event rate of cardiac death, non-fatal reinfarction, or refractory ischaemia. Additional prespecified endpoints included several composites of non-fatal cardiac events (non-fatal reinfarction combined with refractory ischaemia, all recurrent ischaemia, or the need for myocardial revascularisation). The diagnosis of ischaemia, whether refractory or recurrent, and the need for myocardial revascularisation, was always based on objective electrocardiographical evidence of ischaemia, either at rest or on exertion. RESULTS: For the trial primary endpoint, 131 events occurred in the 444 placebo patients and 97 events in the 430 diltiazem patients (hazard ratio 0.79; 95% CI, 0.61-1.02; p=0.07). For non fatal cardiac events, diltiazem treatment was associated with a relative decrease (0.76; 0.58-1.00) in the combined event rate of non-fatal reinfarction and refractory ischaemia. There was a similar decrease in the composite non-fatal endpoints of non-fatal reinfarction combined with all recurrent ischaemia (0.80; 0.64-1.00) and non-fatal reinfarction combined with the need for myocardial revascularisation (0.67; 0.46-0.96). The need for myocardial revascularisation alone was significantly reduced by 42% (0.61; 0.39-0.96). No major safety issues were encountered. CONCLUSIONS: Diltiazem did not reduce the cumulative occurrence of cardiac death, non-fatal reinfarction, or refractory ischaemia during a 6 month follow-up, but did reduce all composite endpoints of non-fatal cardiac events, especially the need for myocardial revascularisation. PMID- 10832826 TI - Favourable and unfavourable effects on long-term survival of radiotherapy for early breast cancer: an overview of the randomised trials. Early Breast Cancer Trialists' Collaborative Group. AB - BACKGROUND: The long-term effects of radiotherapy on mortality from breast cancer and other causes remain uncertain. METHODS: A meta-analysis was done of 10-year and 20-year results from 40 unconfounded randomised trials of radiotherapy for early breast cancer. It involved central review of individual patients' data on recurrence and cause-specific mortality from 20000 women, half with "node positive" disease. Radiotherapy fields generally included not only chest wall (or breast) but also axillary, supraclavicular, and internal mammary nodes. FINDINGS: A reduction of approximately two-thirds in local recurrence was seen in all trials, largely independent of the type of patient or type of radiotherapy (8.8% vs 27.2% local recurrence by year 10). Hence, to assess effects on breast cancer mortality of substantially better local control, results from all trials were combined. Breast cancer mortality was reduced (2p=0.0001) but other, particularly vascular, mortality was increased (2p=0.0003), and overall 20-year survival was 37.1% with radiotherapy versus 35.9% control (2p=0.06). There was little effect on early deaths, but logrank analyses of later deaths indicate that, on average after year 2, radiotherapy reduced annual mortality rates from breast cancer by 13.2% (SE 2.5) but increased those from other causes by 21.2% (SE 5.4). Nodal status, age, and decade of follow-up strongly affected the ratio of breast cancer mortality to other mortality, and hence affected the ratio of absolute benefit to absolute hazard from these proportional changes in mortality. INTERPRETATION: Radiotherapy regimens able to produce the two-thirds reduction in local recurrence seen in these trials, but without long-term hazard, would be expected to produce an absolute increase in 20-year survival of about 2-4% (except for women at particularly low risk of local recurrence). The average hazard seen in these trials would, however, reduce this 20-year survival benefit in young women and reverse it in older women. PMID- 10832827 TI - Trends in trauma care in England and Wales 1989-97. UK Trauma Audit and Research Network. AB - BACKGROUND: In 1988, the Royal College of Surgeons reported major deficiencies in trauma care in UK hospitals. We investigated whether and how that care has changed in the last decade by use of data collected by the UK Trauma Audit and Research Network. METHODS: We analysed injury-severity, process, and outcome variables from 91602 patients' records on the database at the end of 1997, collected from 97 (49% of trauma-receiving) hospitals in England, Wales, and two in Ireland. We did longitudinal analyses of odds of death, process variables, and individual hospitals' performance. We took account of potential selection bias from missing data and recruitment of new hospitals. FINDINGS: The severity adjusted odds of death after trauma declined gradually from 1989 (odds ratio 1997/1989 0.63 [95% CI [0.49-0.82]). In 1997, the reduction in odds of death was significant even after adjustment for missing data (ratio 1997/1989 0.72 [0.55 0.92]) and recruitment of new hospitals (0.64 [0.44-0.93]). There was significant variability in the proportion of survivors (adjusted for severity of injury and age) between the highest and lowest 10% of UK hospitals. The time between the call to the emergency services and arrival at hospital increased from 32 min in 1989 to 45 min in 1997, irrespective of injury severity. The proportion of severely injured patients seen first by senior doctors increased from 32% to 60%. INTERPRETATION: Hospital care has made a valuable but variable contribution to reductions in case fatality after injury in the UK in the past 10 years, though further improvement is possible. PMID- 10832828 TI - Epidemic nephritis in Nova Serrana, Brazil. AB - BACKGROUND: Outbreaks of nephritis have been rare since the 1970s. From December, 1997, to July, 1998, 253 cases of acute nephritis were identified in Nova Serrana, Brazil. Seven patients required dialysis, and three patients died. We did a case-control study to investigate the cause of the outbreak. METHODS: Using a matched cluster design, we examined seven recent patients, their family members (n=23), and members of neighbourhood-matched control households (n=22). We subsequently interviewed 50 patients and 50 matched controls about exposure to various dairy products. We also cultured dairy foods and took udder-swab and milk samples from cows. FINDINGS: Throat cultures indicated that nephritis was associated with group C Streptococcus equi subspecies zooepidemicus, a cause of bovine mastitis. S. zooepidemicus was detected in four of seven case households (six of 30 people) and no control households (p=0.09). Patients were more likely than matched controls to have consumed a locally produced cheese called queijo fresco (matched odds ratio 2.1, p=0.05). The nephritis attack rate was 4.5 per 1000 in Nova Serrana but 18 per 1000 in the village Quilombo do Gaia (p=0.003). The largest supplier of unpasteurized queijo fresco was a farm in Quilombo do Gaia. S. zooepidemicus was not detected in food samples or in swabs collected from cows in August, 1998, although mastitis was evident among cows on the suspected farm. Throat cultures of the two women who prepared cheese on this farm yielded the outbreak strain of S. zooepidemicus. After the cheese was removed from the distribution system, no further cases were reported. INTERPRETATION: A large outbreak of glomerulonephritis was attributed to S. zooepidemicus in unpasteurised cheese. This outbreak highlights the dangers of consuming unpasteurized dairy products and need for global efforts to promote food safety. PMID- 10832829 TI - Association of nonsense mutation of dystrophin gene with disruption of sarcoglycan complex in X-linked dilated cardiomyopathy. AB - BACKGROUND: In a systematic analysis of inherited forms of cardiomyopathy, we previously identified a family with X-linked dilated cardiomyopathy characterised by a mutation in the rod region of dystrophin. We have now attempted to eludicate the genetic mechanism involved in this disease, as well as the role of dystrophin associated glycoproteins. METHODS: The affected dystrophin epitope, which lacks binding to the dys-1 antibody, was analysed by single-strand conformation polymorphism analysis, reverse-transcription PCR, and DNA sequencing. Effects on dystrophin-associated glycoproteins were studied by immunohistochemistry and western blotting. FINDINGS: A translation-termination mutation (C4148T) in exon 29 of the dystrophin gene was found in all affected family members. Alternative splicing rescued the reading frame and led to the expression of a dystrophin molecule lacking 50 aminoacids both in cardiac and skeletal muscle. Immunohistochemical analysis of the dystrophin-associated proteins revealed a reduction of beta-sarcoglycan and delta-sarcoglycan in the sarcolemma of cardiac muscle but not skeletal muscle tissue. However, western blotting revealed similar amounts of sarcoglycan subunits in both tissues. INTERPRETATION: The molecular mechanism of this subtype of X-linked cardiomyopathy may be explained by a conformational change in exon-29-deleted dystrophin, resulting in disruption of the sarcoglycan assembly in heart muscle but not skeletal muscle. PMID- 10832830 TI - An unusual case of acute colitis. PMID- 10832831 TI - Evidence for novel non-angiogenic pathway in breast-cancer metastasis. Breast Cancer Progression Working Party. AB - We have reported that angiogenic primary breast carcinoma can relapse not only as angiogenic, but also as non-angiogenic, lung metastases. This non-angiogenic pathway seems to be a novel pathway of cancer progression, and such tumours are likely to be resistant to antiangogenic treatment. PMID- 10832832 TI - Comparison of trends in prostate-cancer mortality in England and Wales and the USA. AB - Although trends in prostate-cancer screening and disease incidence differ substantially between the USA and England and Wales, trends in mortality are very similar. PMID- 10832833 TI - Presence of therapeutic drugs in the environment. AB - Therapeutic drugs can contaminate the environment because of metabolic excretion, improper disposal, or industrial waste. To assess the extent of this contamination, we listed drugs thought to be putative priority pollutants according to selected criteria, and measured them in Lombardy, Italy. Most drugs were measurable in drinking or river waters and sediments, suggesting that pharmaceutical products are widespread contaminants, with possible implications for human health and the environment. PMID- 10832834 TI - Restoration of thalamocortical connectivity after recovery from persistent vegetative state. AB - By use of H2(15)O positron emission tomography we have shown that functional connectivity between intralaminar thalamic nuclei and prefrontal and anterior cingulate cortices was altered during vegetative state but not after recovery of consciousness. PMID- 10832835 TI - Written information given to patients and families by palliative care units: a national survey. AB - A national survey of written information given to patients and their families by pallative care units found that 64% of leaflets could be understood by only an estimated 40% of the British population. Many of the leaflets did not meet basic guidelines on legibility and readability. PMID- 10832836 TI - Mobile phones: are they safe? PMID- 10832837 TI - Hi-tech breast-cancer screening: will it deliver? PMID- 10832838 TI - Cults in Japan con public with alternative health remedies. PMID- 10832839 TI - Fury in Spain at appointment of controversial government health advisor. PMID- 10832840 TI - Cautious welcome for antiretroviral cost reduction in South Africa. PMID- 10832841 TI - South Africa reels from blood-donation debacle. PMID- 10832842 TI - Jordan starts campaign to tackle high rates of smoking. PMID- 10832843 TI - Violence in public health and preventive medicine. PMID- 10832844 TI - Violence and provision of community psychiatry. PMID- 10832845 TI - Suffer the little children... PMID- 10832846 TI - Violence in research. PMID- 10832847 TI - The non-existent role of injury prevention in medical practice. PMID- 10832848 TI - The ward. PMID- 10832849 TI - Cultural aspects of violence against women in Japan. PMID- 10832850 TI - War. PMID- 10832851 TI - Use of antibiotics in penetrating craniocerebral injuries. "Infection in Neurosurgery" Working Party of British Society for Antimicrobial Chemotherapy. AB - The Working Party was instituted to investigate the rationale of prophylactic and therapeutic antibiotic use in penetrating craniocerebral injuries (PCCI), and to make recommendations for current practice. A systematic review of papers on civilian and military PCCI over the past 25 and 50 years, respectively, was done via electronic databases and secondary sources, and data were evaluated. Guidelines on the removal of indriven bone or metal fragments only if further neural damage can be avoided were supported. However, no publications were identified where the data on infection or its treatment and prevention were complete or satisfactorily derived, and no controlled trials have been published. All studies were retrospective or anecdotal. Working Party recommendations are based on the data available and the professional experience and knowledge of the members. Broad-spectrum antibiotic prophylaxis is recommended for both military and civilian PCCI, Including those due to sports or recreational injuries. PMID- 10832852 TI - MDMA (Ecstasy) neurotoxicity: assessing and communicating the risks. PMID- 10832853 TI - UK and USA breast cancer deaths down 25% in year 2000 at ages 20-69 years. PMID- 10832854 TI - Controlled withdrawal for psychotropic drugs. PMID- 10832855 TI - Fetal surveillance must go on. PMID- 10832856 TI - Colitis associated with docetaxel-based chemotherapy. PMID- 10832857 TI - The heart of psychotropic drug therapy. PMID- 10832858 TI - The heart of psychotropic drug therapy. PMID- 10832859 TI - PET scanning and neuronal loss in acute vegetative state. PMID- 10832860 TI - PET scanning and neuronal loss in acute vegetative state. PROPAC-Study Group. PMID- 10832861 TI - PET scanning and neuronal loss in acute vegetative state. PMID- 10832862 TI - Community care and schizophrenia. PMID- 10832863 TI - Community care and schizophrenia. PMID- 10832864 TI - Breast-cancer treatment put into question. PMID- 10832865 TI - Pacifiers: protection against sudden infant death syndrome. PMID- 10832866 TI - Pellagra outbreak in Kuito, Angola. PMID- 10832867 TI - On environmental lung damage. PMID- 10832868 TI - Online support groups gaining credibility. PMID- 10832869 TI - The body embraces interactivity. PMID- 10832870 TI - Interactions between carbonic anhydrase and its inhibitors revealed by gel electrophoresis and circular dichroism. AB - Structural properties, and especially the differential stability, of complexes between carbonic anhydrase (CA) and three sulfonamide inhibitors, acetazolamide, dorzolamide and methazolamide, were investigated by spectroscopic and electrophoretic techniques. These included denaturant gradient gel electrophoresis either across a urea or a steady-state transverse sodium dodecyl sulfate (SDS) gradient. Acetazolamide, the smallest and most hydrophilic of the sulfonamides, forms the most stable complex in the presence of urea, whereas dorzolamide, with a bulky and hydrophobic structure, is most stable against the effects of SDS. At pH 7.4, complexes with dorzolamide show minimal changes in mobility across the SDS gradient, as if unaffected by the detergent, both in the presence and in the absence of excess ligand in the gel. When bound to both acetazolamide and methazolamide, on the other hand, CA displays an increase in mobility above 0.05% SDS, lower in the presence than in the absence of excess ligand. The finding of a distinct pattern for the unliganded enzyme, however, suggests the complexes can still retain the ligand, although binding of the surfactant changes their charge density. Under saturating conditions and in the presence of SDS, the surface charge of all complexes is much lower than for unliganded, denatured CA. Circular dichroism (CD) spectra clearly indicate that the increase in secondary structure and the decrease in tertiary structure brought about in CA by the presence of low concentrations of SDS are largely prevented by complexing with the inhibitors. These observations point out peculiar properties of each CA inhibitor, of potential value in the definition of their biological activities and also in the potential development of novel antagonist molecules. PMID- 10832871 TI - Double electrophoretic separation and lectin analyses of the component chains of secretory immunoglobulin A from human saliva. AB - A new method is presented for the separation of secretory immunoglobulin A (SIgA) from salivary samples. Salivary proteins (from parotid or stimulated whole mouth saliva) were precipitated with methanol to concentrate SIgA from salivary samples whilst removing other salivary proteins. SIgA purified from breast milk and salivary proteins was separated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under nonreducing conditions. Following completion of electrophoresis the top strip of gel was removed and the proteins present reduced with dithiothreitol. The gel strip was then applied to the top of a second 10% SDS gel, and the proteins were electrophoresed and then stained by Coomassie Brilliant Blue R-250. Three major protein bands were stained in all samples corresponding in molecular mass to secretory component, alpha-heavy chain and light chains of SIgA. Separated proteins were also electroblotted onto nitrocellulose and stained by fluorescein isothiocyanate (FITC). Lectin analysis was then used to detect the O-glycans present on IgA1. Lectins from Helix aspersa and Arachis hypogaea were used to determine the amount of terminal N-acetyl galactosamine and nonsialylated O-glycans, respectively. Maclura pomifera lectin was used to determine the total amount of IgA1 present on the blots. The results indicate that SlgA in stimulated whole mouth saliva, stimulated parotid saliva and purified from breast milk contain similar O-glycans. PMID- 10832872 TI - Cross reactivity between human erythropoietin antibody and horse erythropoietin. AB - Erythropoietin (EPO) is the primary hormone of erythropoiesis. Administration of recombinant human erythropoietin (rhuEPO) to improve racing performance in the horse represents a new form of blood doping, which has been associated with increased mortality. While immunoassay kits have become plentiful, very few commercial hormone assays are made specifically for equine research. There is a strong degree of sequence homology reported for EPO among species, which has allowed antibodies designed for human EPO research to be used to determine EPO concentration in other species. The objective of the present study was to use Western blot analysis to determine whether the antibody to rhuEPO, provided in a commercial radioimmunoassay (RIA) kit, recognizes horse EPO. Western blot analysis of purified rhuEPO and horse plasma was conducted, using the polyclonal goat-antihuman EPO antibody supplied in the Incstar EPO Trac RIA as the primary antibody. Immunoblot analysis revealed a major band at approximately 52 kDa for both rhuEPO and the horse plasma. Our results demonstrate that a human EPO antibody recognizes equine EPO. These findings show that the Incstar EPO Trac RIA hormone assay system can be used to measure equine EPO. PMID- 10832873 TI - Fish muscle parvalbumins as marker proteins for native and urea isoelectric focusing. AB - An isoelectric point (pI) calibration kit containing fish muscle parvalbumins was prepared and tested for its suitability for isoelectric focusing (IEF) in the presence of 8 M urea. The pattern obtained by urea CleanGel IEF consisted of nine bands covering the pI range 4.96-5.64. This range is relevant for species identification of heated fish by urea IEF. The kit may also be used for native IEF in the low pH range, as demonstrated by running an extract made from the kit together with water-soluble fish muscle proteins on Servalyt Precotes 3-6. PMID- 10832874 TI - Gel electrophoretic mobility of single-stranded DNA: the two reptation field dependent factors. AB - The reptation model is the dominant theory in understanding the electrophoretic separation of single-stranded DNA molecules in gels or entangled polymer solutions. Recently, we showed that the Ogston and reptation regimes are separated by an entropic trapping regime at low field intensities. Here, we report the first comparison of the field-dependent part of the DNA mobility for both small and long reptating molecules. We show that both mobilities increase linearly with field intensity, with the mobility of the longer (comigrating) fragments increasing faster than that of the smaller ones. We compare our results to the predictions of the biased reptation model. PMID- 10832875 TI - Assessing microsatellite instability with semiautomated fluorescent technology: application to the analysis of primary brain tumors. AB - The replication error phenotype, revealed by the observation of widespread microsatellite instability (MIN), has been identified as a new mechanism of cancer susceptibility, and the comparison of the allele sizes of polymorphic microsatellite repeats between normal and tumor DNA is now frequently undertaken in colorectal and other human neoplasias. The lack of precise characterization of the electrophoretic profiles of microsatellites is one of the main sources of discord between the rate of MIN reported for the same type of tumor by different investigators. The recent introduction of fluorescent-based semiautomated microsatellite analysis allows a more accurate size comparison, but one or more artificial peaks, generated during polymerase chain reaction (PCR) and/or electrophoresis, are frequently detected along with the true allele peaks. The aim of this study was to characterize the most frequent artificial extra peaks in the short tandem repeats (STRs) used by us to assess MIN in human cancers. We analyzed eight microsatellite loci in 113 primary brain tumors. HumFibra/FGA exhibited the most frequent extra peak formation. For each microsatellite there is a characteristic pattern of artifact formation which must be recognized to avoid a false-positive diagnosis of MIN. PMID- 10832876 TI - Sequencing and four-state minisatellite variant repeat mapping of the D1S7 locus (MS1) by fluorescence detection. AB - Minisatellite variant repeat mapping by polymerase chain reaction (MVR-PCR) reveals an enormous degree of variation in the human minisatellite regions. The original approach involved the use of 32P-labelled probes to detect the MVR-PCR products generated. To date, the loci mapped include D1S8, D7S21 and D16S309. However, the most polymorphic locus, D1S7 (MS1), which has been used in forensic analysis, has presented technical difficulties, initially due to its short 9 bp repeats that are much shorter than any conventional primer sequences. This was overcome by using the method of "wrapping around" primers employing inosine at the redundancy position. The difficulty of cloning highly repetitive DNA was overcome by utilising specialised competent SURE cells. We report the cloning and sequencing of selected short MS1 alleles to determine the variety of repeat types. This survey revealed nine types, four of which represented greater than 80% of the sequenced repeats. The reported MVR-PCR system maps the MS1 locus for these four common repeat types by fluorescence detection. PMID- 10832877 TI - Identification of hallucinogenic fungi from the genera Psilocybe and Panaeolus by amplified fragment length polymorphism. AB - Unambiguous identification of the hallucinogenic fungi of the genera Psilocybe and Panaeolus is required by national and international drug control legislation. We report on a DNA-based test using the technique of amplified fragment length polymorphism (AFLP). AFLP can differentiate species of the two genera Psilocybe and Panaeolus by using different primer sets. The identification of hallucinogenic fungi using a DNA-based test, which can be used in conjunction with morphological features, will assist in forensic investigations. PMID- 10832878 TI - Voltage-induced release of nucleic acids from palaeontological samples. AB - Most of the protocols for the recovery of ancient DNA from palaeontological specimens are time-consuming and tend to yield inconsistent polymerase chain reaction (PCR) results. "Voltage-induced release" is a novel and rapid approach for the extraction of ancient DNA. Nucleic acids are directly electrophoresed out of powder derived from hard and soft tissues. This technique is much faster than other methods in which pulverized tissue conventionally undergoes time-consuming crude lysis steps. The total preparation time is 5-6 h. The reliability of the voltage-induced release method was validated by (i) measuring the ratio of D-to L enantiomers of the amino acids aspartic acid, alanine, and leucine, and (ii) by specific PCR amplification of four single-copy markers of human chromosome 17 and 18. We compare voltage-induced release to a frequently used silica-based protocol. DNA extracted employing voltage-induced release was more effective in PCR amplifications, which may be attributed to the effective removal of PCR inhibitors. PMID- 10832879 TI - From small charged molecules to oligomers: a semiempirical approach to the modeling of actual mobility in free solution. AB - According to Stokes' treatment, the ionic mobility of particles, which are small with respect to Debye length, is usually considered to be proportional to the nominal charge and inversely proportional to the hydrodynamic radius. Experimentally, it is well known, however, that the ionic mobility of a small multicharged molecule does not depend linearly on its nominal charge in a wide range. This behavior can be accounted for by a condensation of the charge or a modification of the friction coefficient with the charge. This paper presents a semiempirical modeling of the actual mobility based on the assumption of additivity of frictional contributions pertaining to the uncharged molecular backbone and to each charged or uncharged moiety. Condensation of the charge was not considered. The model first appeared to be suitable for multicharged analytes having a characteristic dimension smaller than the Debye length, such as benzene polycarboxylic acids and polysulfated disaccharides. This approach was then adapted to account for the actual mobilities of singly and evenly charged oligomers (N-mers) having a dimension smaller than or similar to the Debye length. Rather good experimental agreement was obtained for polyalanines and polyglycines (N < or = 6), fatty acid homologs, fully sulfonated polystyrene oligomers (N < or = 13), and polycytidines (N < or = 10). Especially the influence of the polymerization degree on the mobility of oligomers having identical charge densities was clarified. It is also shown that the electrophoretic contribution to the overall friction coefficient increases linearly with the nominal charge but hardly depends on the chemical nature of the charged moieties. This model should be of interest to evaluate the role of various physicochemical phenomena (hydrodynamic and electrophoretic frictions, hydrodynamic coupling, charge condensation) involved in the migration of charged oligomers. PMID- 10832880 TI - Diffusion coefficient and capacity factor in capillary electrokinetic chromatography with replaceable charged polymeric pseudophase. AB - Apparent diffusion coefficients, Dapp,i, were determined in solutions with a polycationic additive -- poly(diallyidimethylammonium) -- acting as a pseudostationary phase for electrokinetic chromatography. They were determined for six small neutral analytes at five concentrations of the polymeric additive (between 0 and 4% w/w) by a stopped migration method. The apparent diffusion coefficients decrease with increasing polymer concentration only within 40% maximum, an effect that cannot be associated with the macroscopic viscosity of the polymer solution (which increases by a factor of 10). The change of the apparent diffusion coefficients is related to the interaction of the neutral analyte molecules with the polyelectrolyte chain. Applying the model of analyte partitioning between "free" solution and polymer, capacity factors and partition constants were derived from the slope of the 1/Dapp,i vs. polymer concentration curves. Partition constants determined by this method (ranging between 40 and 170) agree with those obtained by electrokinetic chromatography. PMID- 10832881 TI - Affinity capillary electrophoretic studies of complexation between dextrin oligomers and polyiodides. AB - Capillary electrophoresis (CE) has been applied to the study of complexation between dextrins and polyiodides. A baseline separation of fluorescently labeled dextrin oligomers has provided a unique platform for the observation of a contribution of single oligomers to the complexation process that could previously be measured only in bulk. The complex formation was easily recognized through comparison of peak migration times and peak shapes in the presence and absence of polyiodides. The degree of polymerization (DP) number was found crucial in the binding process, but the I2/I- ratio in a solution also appeared to determine the nature of complexation. The effects of buffer pH and ionic strength upon complexation were also briefly investigated. Diodearray spectra in the visible wavelength range confirmed the differential complexation of unlabeled maltodextrins with different DP values after a CE iodine affinity separation. 13C nuclear magnetic resonance (NMR) spectral data on differently sized dextrin fractions were found to be in good agreement with the results from CE measurements. PMID- 10832882 TI - A new absorbed coating for DNA fragment analysis by capillary electrophoresis. AB - A fully automated coating procedure was devised based on adsorption of a new polymer, poly(dimethylacrylamide-co-allyl glycidyl ether) onto the capillary surface. The whole procedure takes less than 30 min and does not require the use of organic solvents, viscous solutions, or elevated temperature. The coating is stable even under harsh conditions such as alkaline pH, elevated temperature, and denaturant conditions that destroy most other presently available adsorbed coatings. This new adsorbed coating is highly stable and easy to produce in quantity, making it quite unique, and further making it possible to operate with any DNA sieving matrix. Finally, the above-mentioned properties facilitate coating regeneration by a simple wash with a strongly alkaline solution, thus extending the lifetime of the capillary, a highly desirable property for any coating used in biopolymer separations. PMID- 10832883 TI - A new capillary electrophoresis end-column amperometric detection system without the need for capillary/electrode alignment. AB - A self-aligning end-column amperometric detection system for capillary electrophoresis was constructed. In this system, the electrode and capillary were exchanged easily and the capillary/electrode alignment procedure is not required. Gold, gold/mercury amalgam, copper and carbon fiber could be used as the working electrode. The principle is in the use of two disk holders with the capillary and the electrode in the center, so that by inserting the disk holders into a groove in the working electrode port, the capillary and the electrode are automatically aligned and the distance between the capillary and the electrode is assured at 0.24 mm. The relative standard deviation obtained using five different gold/mercury amalgam microdisk electrodes for determination of cysteine was 1.5% for the migration time and 3.3% for the electrophoretic peak current. The simple and convenient system was attractive for the routine analysis by capillary electrophoresis with electrochemical detection. The system was applied to the determination of promethazine hydrochloride in human serum. PMID- 10832884 TI - Monitoring myoglobin by capillary zone electrophoresis with end-column amperometric detection. AB - Capillary zone electrophoresis was employed for the determination of myoglobin in human urine using end-column amperometric detection with a carbon fiber microelectrode at a constant potential of 1.80 V vs. saturated calomel electrode (SCF). The optimum conditions of separation and detection are: 3.73 x 10-4 mol/L sodium diethyl malonyl urea (barbitone sodium), 1.34 x 10-4 mol/L HCl for the buffer solution, 20 kV for separation voltage, 5 kV and 5 s for injection voltage and injection time, respectively. The limit of detection is 4.4 x 10-8 mol/L or 84 amole signal to noise (S/N = 2). The relative standard deviation is 2.9% for the migration time and 2.5% for the electrophoretic peak current. The method can be used for the determination of myoglobin in human urine. The samples can be directly injected and need no pretreatment. The method is also rapid, less than 2 min, and has a recovery rate of 94-106%. PMID- 10832885 TI - Capillary electrophoresis/electrochemical detection system with on-line deoxygenation. AB - A capillary electrophoresis (CE)/electrochemical detection system with on-line deoxygenation was developed, consisting of a deoxygenation injector, a deoxygenation protector, and an electrochemical detection cell. When the system was utilized for 60 min, the steady-state current of oxygen detected could be dropped to 3% of the original value for the gold/mercury amalgam electrode and to 8% of the original value for the gold electrode, and the limit of detection could be decreased two orders of magnitude for the reducible analytes such as TI+ (from 3.1 x 10-5 mol/L to 8.0 x 10-7 mol/L) and metronidazole (from 3.8 x 10-5 mol/L to 4.0 x 10-7 mol/L). PMID- 10832886 TI - Intra- and interinstrument reproducibility of migration parameters in capillary electrophoresis for substance identification in systematic toxicological analysis. AB - The intra- and interinstrument reproducibilities of four capillary electrophoresis instruments were studied for identification purposes in systematic toxicological analysis (STA). A test set of 20 acidic test compounds and 5 reference compounds were analyzed for five days on each instrument using capillary zone electrophoresis (CZE) and micellar electrokinetic chromatography (MEKC). The buffers consisted of 90 mM borate set at pH 8.4 (CZE) and 20 mM phosphate and 50 mM sodium dodecyl sulfate set at pH 7.5 (MEKC). All analyses were carried out using fused silica capillaries at an electric field strength of 52.6 kV/m. The use of a reproducible identification parameter is very important in STA. To deal with the poor reproducibility of the migration time, we recently introduced the corrected effective mobility. In this study, we investigated the intra- and interinstrument reproducibility of the migration time, the effective mobility, and the corrected effective mobility. Large differences in intra instrument reproducibility were found when the migration time was used. The calculation of the effective mobility and the corrected effective mobility diminished these differences and enhanced the interinstrument reproducibility roughly by a factor 3. For (corrected) effective mobilities, intrainstrument reproducibilities were between 0.8-2.6% and interinstrument reproducibilities were between 3.2-3.9%. PMID- 10832887 TI - Capillary electrophoresis in the evaluation of ischemic injury: simultaneous determination of purine compounds and glutathione. AB - An understanding of tissue energy metabolism and antioxidant status is of major interest in the field of organ preservation for transplantation. Nucleotide and glutathione are indicators of cell damage occurring during ischemia and reperfusion. A high performance capillary electrophoresis (HPCE) method with UV detection (185 nm) for the simultaneous analysis of intracellular free ribonucleotides, nucleosides, bases and glutathione (oxidized and reduced form) in myocardial tissues is described. The method does not involve thiol derivatization. The separations were carried out in an uncoated fused-silica capillary, 60 cm long, 52.5 cm to detector, 75 microm ID, with 20 mM Na-borate buffer, pH 10.00, at 20 kV voltage and reading at 185 nm. Injection was hydrostatic for 12 s and total analysis time was 20 min. The technique enables optimum separation of all the compounds examined and has a resolution similar to that of HPLC analysis, with the advantage of fast simultaneous measurement of cell nucleotide metabolism and redox state, not possible with HPLC. PMID- 10832888 TI - Micellar electrokinetic chromatography for the analysis of cefpirome in microdialysis and plasma samples obtained in vivo from human volunteers. AB - Pharmacokinetics of drugs in the human interstitial space fluid can be monitored by means of microdialysis. However, the small-volume microdialysis samples containing low drug concentrations require a sensitive analytical method. In the present study, micellar electrokinetic chromatography (MEKC) is described for the quantification of cefpirome in human microdialysis and plasma samples. Sample preparation of human plasma samples by ultracentrifugation was suitable for comparison of plasma and microdialysate concentrations. Limits of quantification were 2 microg/mL and 0.3 microg/mL for plasma and microdialysate samples, respectively. The limit of detection (LOD) was estimated at 0.2 microg/mL for the plasma and microdialysate samples. In conclusion, MEKC is a reliable and reproducible technique for measuring cefpirome concentrations in microdialysates as well as centrifuged plasma samples. PMID- 10832889 TI - Enantiomeric separation of a group of chiral dihydropyridines by electrokinetic chromatography. AB - Electrokinetic chromatography (EKC) was employed to achieve the enantiomeric separation of a group of chiral 1,4-dihydropyridines (DHPs) with pharmacological activity. Micelles of bile salts alone or mixed with neutral cyclodextrins, micelles of sodium dodecyl sulfate (SDS) mixed with neutral cyclodextrins, and anionic cyclodextrin derivatives, i.e., carboxymethyl-gamma-cyclodextrin (CM gamma-CD), carboxymethyl-beta-cyclodextrin (CM-beta-CD), and succinylated beta cyclodextrin (Succ-beta-CD), were employed as pseudostationary phases. The enantiomeric separation ability of these chiral selectors with respect to DHPs was studied in different experimental conditions. CM-beta-CD was shown to be the best chiral selector to perform the enantiomeric separation of DHPs by EKC. Next, the influence of the CM-beta-CD concentration, the pH and nature of the buffer, the temperature, and the applied voltage on the enantiomeric resolution of DHPs was studied. The use of a 50 mM ammonium acetate buffer, pH 6.7, 25 mM in CM-beta CD together with an applied voltage of 15 or 20 kV, and a temperature of 15 degrees C enabled the individual enantiomeric separation of twelve DHPs, each one into its two enantiomers, and their separation in multicomponent mixtures of up to six DHPs into all their enantiomers. PMID- 10832890 TI - Capillary electrochromatography for pesticide analysis: effects of environmental matrices. AB - An insecticide, pirimicarb, and a fungicide, azoxystrobin, were analyzed by capillary electrochromatography. Nine environmental matrices derived from soil, plant and animal extracts were used. After a series of 311 consecutive injections with no washing between injections, retention times increased by ca. 0.5 min. The use of exaggerated application rates for metabolism studies enabled the detection of xenobiotic pesticide degradates by UV absorbance. A 1.2 mm path length, high sensitivity flow cell gave a gain in sensitivity; however, a further increase in sensitivity of at least two orders of magnitude is required for pesticide residue analysis. Analyte stacking using large volume injections of aqueous samples led to a large increase in retention times. PMID- 10832891 TI - Qualitative determination of urinary morphine by capillary zone electrophoresis and ion trap mass spectrometry. AB - A rapid, sensitive method for the determination of morphine and amphetamine was developed using capillary zone electrophoresis coupled with electrospray interface (ESI), ion-trap tandem mass spectrometry (CE-ESI-MS2). Morphine and amphetamine were separated in 20 mM ammonium acetate buffer (pH 6.6) and detected by ion-trap mass detector in different analytical time segments (0-6.25 min for amphetamine and 6.25-12.0 min for morphine) in which the tune file for each compound was used separately. Molecular ions of morphine (m/z 286) and amphetamine (m/z 136) were detected at 5.77 and 6.83 min, respectively, while product ions of MS2 for each compound (m/z 229, 201 for morphine and m/z 119 for amphetamine) were detected almost exactly at the same time with their parent compounds. The limits of detection (LOD) for MS2 determination were 30 and 50 ng/mL for amphetamine and morphine, respectively, with an S/N ratio of 3. For more sensitive detection of morphine, the sample was injected for a longer time (i.e., 80 s) and hydrodynamically transported into a CE capillary for MS detection. Morphine and its product ion appear at 0.36 and 0.39 min on the ion chromatogram, respectively, with a five-fold increase of detection sensitivity (LOD, 10 ng/mL). The CE-MS system thus established was further applied for forensic urine samples screened as morphine-positive by fluorescence polarization immunoassay (FPIA). These results indicated the feasibility of CE-ESI-MS2 for confirmative testing of morphine in urine sample. PMID- 10832892 TI - Cellulases from the fungi Phanerochaete chrysosporium and Trichoderma reesei as chiral selectors in capillary electrophoresis: applications with displacer plugs and sample preconcentration. AB - The cellulases CBH 58 from the fungus Phanerochaete chrysosporium and CBH I from the fungus Trichoderma reesei were compared as chiral selectors in capillary electrophoresis (CE) applying the partial filling technique. Amines, e.g., norephedrine, two bambuterol analogs, as well as acids, e.g., di-p-toluoyl tartaric acid and dibenzoyl tartaric acid, which could not be enantioseparated in the liquid chromatographic use of the selectors, could be separated in the corresponding CE experiments. Due to the very high enantioselectivities, terbutaline, alprenolol and propranolol could be completely enantioresolved with selector plugs shorter than the sample plugs. The affinity of propranolol to CBH 58 was so high at pH 7.0 that neither of the enantiomers reached the detector; therefore, a plug of the displacing disaccharide cellobiose was injected after the sample to elute the propranolol enantiomers. The enantiomers could also be made to leave the capillary at opposite ends, thereby causing an infinite enantioresolution. A new preconcentration technique was introduced, which takes advantage of the very high affinity of propranolol to CBH 58 and the eluting ability of cellobiose. A 12.5 cm long plug of rac-propranolol could be preconcentrated and enantioseparated in a single procedure. PMID- 10832894 TI - Determination of S-nitrosoglutathione in erythrocytes by capillary zone electrophoresis. AB - A method for separation and quantification of S-nitrosoglutathione in red cell extracts by capillary electrophoresis is reported. The method is based on the direct analysis of the metaphosphoric acid erythrocyte extract containing diethylenetriaminepentaacetic acid. Optimization of the method is briefly discussed. Best results in the shortest time were obtained at 25 degrees C, using a coated capillary, 7 kV applied voltage and phosphate sodium 40 mmol/L (pH 2.2) as running buffer. Reproducibility, detection limits, and recoveries of S nitrosoglutathione analyses were checked. The results evidenced that S nitrosoglutathione is formed in erythrocytes treated with S-nitrosocysteine, a transnitrosating agent. Under our experimental conditions, the contemporaneous detection and quantification of reduced and oxidized glutathione present in cell extract could also be performed. PMID- 10832893 TI - Chiral separation with dipeptide-terminated polymeric surfactants: the effect of an extra heteroatom on the polar head group. AB - Chiral recognition of two binaphthyl derivatives and three benzodiazepines were studied by use of polymeric surfactants in electrokinetic chromatography. Four specific dipeptide terminated (multichiral) micelle polymers were synthesized for this study. These include poly (sodium-N-undecanoyl-L-alanyl-leucinate)-(poly L SUAL), poly (sodium-N-undecanoyl-L-valyl-leucinate) (poly L-SUVL), poly (sodium-N undecanoyl-Lseryl-leucinate) (poly L-SUSL), and poly(sodium-N-undecanoyl-L threonyl-leucinate) (poly L-SUTL). In addition to the chiral separation study, the physicochemical properties (critical micelle concentration and specific rotation) of each polymer were investigated. The molecular weights of the various dipeptide-terminated micelle polymers were determined using analytical ultracentrifugation. These dipeptide-terminated micelle polymers were designed to study the effect of the extra heteroatom at the polar head group of the micelle polymer (i.e., poly L-SUSL compared to poly L-SUAL and poly L-SUTL compared to poly L-SUVL) on the enantiomeric separation of the binaphthyl derivatives and benzodiazepines. The synergistic effect of three chiral centers (poly L-SUTL) provided improved resolution over that of two chiral centered dipeptide terminated micelle polymer in the case of (+/-)-temazepam, (+/-)-oxazepam, (+/-) binaphthol, and (+/-)-binaphthol phosphate. The chiral recognition mechanisms in these cases were additionally controlled by the presence of the extra heteroatom located on the polar head group of the micelle polymers. PMID- 10832895 TI - Simultaneous chiral separation of triadimefon and triadimenol by sulfated beta cyclodextrin-mediated capillary electrophoresis. AB - Enantiomeric separation of two triazole fungicides, triadimefon and triadimenol, was investigated in sulfated beta-cyclodextrin (sulfated beta-CD)-mediated capillary electrophoresis (CE) systems. It was found that, at pH 2-4, sulfated beta-CD exhibited strong chiral recognition towards both triadimefon and triadimenol. The enantiorecognition was believed to result from the multiple interactions between sulfated beta-CD and the analytes, which included inclusion effect, electrostatic interaction, and hydrogen bonding. Under optimal conditions (phosphate buffer with 2% sulfated beta-CD, pH 2.5), simultaneous resolution of all chiral isomers of triadimefon and triadimenol was achieved in less than half an hour. In conjunction with solvent extraction and subsequent enrichment by solid-phase extraction (SPE), this new enantioseparation method was applied successfully in the study of stereoselectivity associated with the biotransformation of triadimefon to triadimenol by soil microorganisms. The present methodology was superior to the commonly adopted chiral gas chromatography (GC) approach in that a very mild procedure was involved from sample extraction to the ultimate chiral separation. Thus, the disturbance of the enantiomeric distribution patterns of the original soil samples by heat stress was an unlikely scenario. Furthermore, it was discovered that, owing to the unique selectivity of the present separation strategy, there was virtually no interference from the soil matrix, which led to improvements in both sensitivity and selectivity in real sample determination. PMID- 10832896 TI - Preparation and characterization of calixarene-coated capillaries for capillary electrophoresis. AB - Preparation and characterization of calixarene-coated capillaries for capillary electrophoresis (CE) were exemplified with p-allylcalix[4]arene (pACX4) which was immobilized to the fused silica surface using gamma-methacryloxypropyl trimethoxysilane (gamma-MAPS) as linking agent. Successful gamma-ACX4 coating was suggested by the greatly decreased electroosmotic flow (EOF), due to the introduction of phenolic hydroxyl groups on the inner surface of the capillaries. A slight slope of EOF versus pH at pH <8 would help make the separation reproducible. The coated columns also featured a low ultraviolet (UV) absorption background and long lifetime (> 6 months at 4 or = 18 years) describing the epidemiology of smokeless tobacco use in the southeastern United States. These studies indicate that the Southeast, with its strong ties to tobacco production, is an area that requires intensive tobacco cessation strategies targeting the use of smokeless tobacco as well as cigarettes. PMID- 10832941 TI - Perpetrators of abusive head trauma: a comparison of two geographic populations. AB - BACKGROUND: Abusive head trauma accounts for significant morbidity and mortality in infants. We compared a Southern population of victims with those in a previous study of a Western population, which found that men, particularly fathers and mothers' boyfriends, are the most common perpetrators. METHODS: All cases of child abuse identified in a teaching hospital were prospectively reviewed for cases of abusive head trauma, and the perpetrators were identified. RESULTS: Of the 76 cases of head trauma identified, 27 met the criteria for the study. The demographics of the perpetrators closely match those of the Western group. Men are the predominant perpetrators, with fathers committing 45% and boyfriends 25% of these injuries. CONCLUSIONS: Despite the differences in study design and population demographics, men are the most common perpetrators of abusive head trauma in both populations. PMID- 10832942 TI - Vascular injuries of the upper extremity. AB - Vascular injuries of the upper extremity represent approximately 30% to 50% of all peripheral vascular injuries. The majority of injuries are to the brachial artery, and 90% of injuries are due to penetrating trauma. Return of function is often related to concomitant injury to peripheral nerves. However, timely restoration of blood flow is essential to optimize outcome. The diagnosis is made by physical examination and limited Doppler ultrasonography. Arteriography may be helpful if there are multiple sites of injury. Anticoagulation with heparin should be given if not otherwise contraindicated. Revascularization should be completed within the critical ischemic time: 4 hours for proximate injuries and 12 hours for distal injuries. Revascularization methods include resection and primary repair or resection with an interposition graft. The sequence of repair of multiple injuries to the extremity begins with arterial revascularization followed by skeletal stabilization and nerve and tendon repair. PMID- 10832943 TI - Use of HFE mutation analysis for hereditary hemochromatosis: the need for physician education in the translation of basic science to clinical practice. AB - BACKGROUND: Hereditary hemochromatosis (HH) is a common hereditary disorder of iron metabolism causing iron overload, organ failure, and malignancy. Preclinical diagnosis using HFE gene analysis followed by prophylactic phlebotomy can completely prevent the disease. METHODS: We conducted a mail survey of all registered primary care physicians, gastroenterologists, and hematologists in Arkansas (n = 860) to determine utilization of HFE mutation analysis in clinical medicine a year after the new molecular test first became available. RESULTS: Of 346 responding physicians (40%), 71 (21%) were aware of the test, 36 (10%) knew that the test was available in Arkansas, and 10 (3%) had used the test. One physician had used the test to screen first-degree relatives of a homozygous HH proband. CONCLUSIONS: Because of poor utilization of the test, the discovery of the role of HFE mutations in HH has had minimal impact on clinical care in Arkansas. PMID- 10832944 TI - Medical students' attitudes toward pain and the use of opioid analgesics: implications for changing medical school curriculum. AB - BACKGROUND: Barriers to pain management include physicians' lack of knowledge and attitudes. Our aim was to investigate future physicians' knowledge and attitudes toward pain and the use of opioid analgesics. METHODS: We tested a medical school class during their freshman and senior years. Stepwise regression analysis was used to identify the personal traits that predicted opiophobia. RESULTS: The professionalization process of medical training may reinforce negative attitudes. Psychologic characteristics were associated with reluctance to prescribe opioids, and fears of patient addiction and drug regulatory agency sanctions. CONCLUSIONS: Consistent attitudes were found in senior medical students with preferences for certain specialty areas and the practitioners of their future specialties, suggesting a "preselection" effect. Higher scores on reliance on high technology, external locus of control, and intolerance of clinical uncertainty were associated with higher scores on one or more of the three dimensions of opiophobia. Implications for medical education are discussed. PMID- 10832945 TI - Physicians' attitudes toward pain and the use of opioid analgesics: results of a survey from the Texas Cancer Pain Initiative. AB - BACKGROUND: Despite extensive progress in the scientific understanding of pain in humans, serious mismanagement and undermedication in treating acute and chronic pain is a continuing problem. This study was designed to examine the barriers to adequate pain management, especially as they might be associated with community size and medical discipline. METHODS: A 59-item survey was used to measure physicians' attitudes, knowledge, and psychologic factors that contribute to pain management practices. RESULTS: Overall, a significant number of physicians in this survey revealed opiophobia (prejudice against the use of opioid analgesics), displayed lack of knowledge about pain and its treatment, and had negative views about patients with chronic pain. There were significant differences among groups of physicians based on size of geographic practice area and medical discipline. CONCLUSIONS: New educational strategies are needed to overcome these barriers and to improve pain treatment in routine medical practice. The effect of practice milieu must be taken into consideration. PMID- 10832946 TI - Quinolones and tendon ruptures. AB - We report two cases of tendon rupture associated with ciprofloxacin. One patient had a complete rupture of an Achilles tendon 6 months after taking the medication. The other case involved a partial rupture of the subscapularis tendon. Both ruptures occurred with minimal mechanical stress on the tendons, suggesting that the fluoroquinolone increased the susceptibility to rupture. We also review the literature describing the association between fluoroquinolones and tendon rupture and discuss the mechanisms explaining the heightened risk of tendon rupture associated with these drugs. PMID- 10832947 TI - Fatal bladder cancer and dermatomyositis. AB - Dermatomyositis is an uncommon inflammatory myopathy accompanied by characteristic cutaneous findings. Adult-onset dermatomyositis is often associated with internal malignancy. We report a case of dermatomyositis associated with an aggressive and fatal case of transitional cell carcinoma of the bladder. PMID- 10832948 TI - Pulmonary sarcoidosis: presentation as bilateral spontaneous hydropneumothorax and pulmonary infiltrates. AB - Pulmonary manifestations in sarcoidosis vary, ranging from asymptomatic chest radiographic abnormalities to progressive destruction of lung parenchyma with respiratory insufficiency. We describe a case of sarcoidosis in a patient with bilateral hydropneumothorax, parenchyma infiltrates, and respiratory insufficiency. Hydropneumothorax is extremely rare, and to our knowledge only two cases have been reported. PMID- 10832949 TI - Tracheal lipoma: a rare intrathoracic neoplasm. AB - Primary tracheal lipomas are extremely rare neoplasms. The typical patient is a middle-aged man with complaints of cough and shortness of breath. Often, the diagnosis is delayed, and patients are treated for asthma or bronchitis. The diagnosis of a tracheal lipoma is best approached by computed tomography (CT) and bronchofibroscopy. Tracheobronchial lipomas may be successfully excised endoscopically or by laser therapy. Open surgical resection is required when the lipoma extends extraluminally. PMID- 10832950 TI - Retrograde jejunogastric intussusception. AB - Retrograde gastrointestinal intussusception is a rare entity, most commonly reported after gastric resection and gastrojejunostomy. Its occurrence in the absence of previous gastric resection is extremely unusual, with only four cases reported. All cases were associated with previously placed gastrostomy tubes and implicated these as the inciting factor. We present a fifth case and review the literature. The mechanism of this phenomenon is described and recommendations to prevent this potentially fatal complication are made. PMID- 10832951 TI - Dual-energy x-ray absorptiometry in osteopetrosis. AB - We have used dual-energy x-ray absorptiometry (DXA) in evaluation and follow-up of a patient with osteopetrosis, before and after cord blood transplantation. Other methods of follow-up in such cases have been described, but the use of DXA has not previously been reported. We have shown that DXA offers a safe means of assessing disease progression, the timing of treatment, and response after therapy for osteopetrosis. PMID- 10832952 TI - Hashimoto's encephalopathy. AB - Hashimoto's encephalopathy is a subacute condition associated with autoimmune thyroiditis. Its presentation varies from focal neurologic deficits to global confusion. Unlike encephalopathy associated with hypothyroidism, Hashimoto's encephalopathy responds to steroid therapy and not thyroxine replacement. PMID- 10832953 TI - Delayed death from pulmonary tuberculosis: unsuspected subtherapeutic drug levels. AB - A patient with fulminant pulmonary tuberculosis died after 41 days of intensive care despite pansensitive organisms and no known underlying immunosuppression. Two factors leading to death in this patient were a delay in seeking medical attention and a subtherapeutic serum level of rifampin, though no obvious evidence of malabsorption existed. Malabsorption of antitubercular drugs is under recognized and of extreme importance in the treatment of critically ill patients with active pulmonary tuberculosis. Factors associated with mortality from tuberculosis and selected aspects of critical care management are discussed. PMID- 10832954 TI - Acute abdomen with colonic necrosis induced by Kayexalate-sorbitol. AB - Colonic necrosis is an unusual complication after treatment of hyperkalemia with sodium polystyrene sulfonate (SPS, Kayexalate) in sorbitol. To increase awareness of this complication, we report a case of necrosis of the transverse colon in a patient given oral and rectal SPS-sorbitol for hyperkalemia. Colonic necrosis was manifested as an acute abdomen within 24 hours of initial administration. Prompt surgical resection of the necrotic transverse colon permitted rapid recovery of bowel function. Although SPS crystals are seen microscopically in the necrotic bowel, experimental evidence implicates the sorbitol component of the SPS sorbitol in the pathogenesis of colonic necrosis. A high index of suspicion for the unusual complication of colonic necrosis after oral or rectal administration of SPS-sorbitol may allow prompt recognition and surgical cure. PMID- 10832955 TI - Trovafloxacin-induced weakness due to a demyelinating polyneuropathy. AB - Fluoroquinolones have been associated with peripheral sensory disorders and weakness, especially in patients with underlying myasthenia gravis or myasthenia like Eaton-Lambert syndrome. Trovafloxacin is a relatively new quinolone for which these side effects have not been described. We report a case of diffuse weakness due to a demyelinating polyneuropathy that began after initiation of trovafloxacin in a patient without an underlying neurologic disorder. PMID- 10832956 TI - Atypical presentation of hepatocellular carcinoma. AB - Hepatocellular carcinoma is relatively rare in the United States, and the patterns of extrahepatic manifestations are diverse. Disease dissemination occurs through hematogenous routes to frequently involve the lungs, bone, adrenal glands, and pancreas. Soft tissue metastasis is extremely rare and mandates systematic pathologic analysis, which may include the use of immunohistochemical staining for tumor-specific markers. Relevant tumor markers that can assist in localizing the site of origin for adenocarcinoma include carcinoembryonic antigen, alpha-fetoprotein, vimentin, and anticytokeratins. We detail the utility of immunohistochemistry in evaluating tumors of unknown origin. PMID- 10832957 TI - Fluoroquinolones. PMID- 10832958 TI - Lessons from recent hypertension trials: how low to lower blood pressure. PMID- 10832959 TI - Systolic blood pressure, diastolic blood pressure, or pulse pressure: which is more important for the practicing clinician? PMID- 10832960 TI - Combination agents as a means of improving adherence and other aspects of the sixth Joint National Committee Report. PMID- 10832961 TI - Evaluation and treatment of renovascular disease in the elderly: clues for the clinician. PMID- 10832962 TI - Treating the African American with hypertension: does race really matter? PMID- 10832963 TI - Dietary Bifidobacterium lactis (HN019) enhances resistance to oral Salmonella typhimurium infection in mice. AB - The ability of a newly identified probiotic lactic acid bacterial strain, Bifidobacterium lactis (HN019), to confer protection against Salmonella typhimurium was investigated in BALB/c mice. Feeding mice with B. lactis conferred a significant degree of protection against single or multiple oral challenge with virulent S. typhimurium, in comparison to control mice that did not receive B. lactis. Protection included a ten-fold increase in survival rate, significantly higher post-challenge food intake and weight gain, and reduced pathogen translocation to visceral tissues (spleen and liver). Furthermore, the degree of pathogen translocation showed a significant inverse correlation with splenic lymphocyte proliferative responses to mitogens, blood and peritoneal cell phagocytic activity and intestinal mucosal anti-S. typhimurium antibody titers in infected mice; all of these immune parameters were enhanced in mice fed B. lactis. Together, these results suggest that dietary B. lactis can provide a significant degree of protection against Salmonella infection by enhancing various parameters of immune function that are relevant to the immunological control of salmonellosis. Thus dietary supplementation with B. lactis provides a unique opportunity for developing immune-enhancing probiotic dairy food products with proven health benefits. PMID- 10832964 TI - Phylogenic and phenotypic characterization of some Eubacterium-like isolates from human feces: description of Solobacterium moorei Gen. Nov., Sp. Nov. AB - Three isolated strains from human feces were characterized by biochemical tests and 16S rDNA analysis. Phylogenetic analysis revealed that these isolated strains were members of the Clostridium subphylum of gram-positive bacteria. The phenotypic characters resembled those of the genus Eubacterium, but these strains were shown to be phylogenetically distant from the type species of the genus, Eubacterium limosum. The strains showed a specific phylogenetic association with Holdemania filiformis and Erysipelothrix rhusiopathiae. Based on a 16S rDNA sequence divergence of greater than 12% with H. filiformis and E. rhusiopathiae, a new genus, Solobacterium, is proposed for three strains, with one species, Solobacterium moorei. The type strain of Solobacterium moorei is JCM 10645T. PMID- 10832965 TI - Chemical and serological properties of lipopolysaccharides from Vibrio parahaemolyticus O-untypeable strains isolated from patients. AB - Chemical and serological studies have been carried out on the O-antigenic lipopolysaccharides (LPS) of six strains, U-6443, W-90144, X-3972, AD-7999, 90A 6611 and KX-V212, of Vibrio parahaemolyticus isolated from patients. The O serotypes of these strains have not been identified because they were not agglutinated by any diagnostic antisera against known O-serotype strains. A compositional sugar analysis of their LPS revealed that out of the six O untypeable (OUT) strains, U-6443, W-90144 and AD-7999 strains belonged to chemotype II (chemotype of O2), 90A-6611 and KX-V212 strains to chemotype III (chemotype of O3, O5, O11 and O13) and X-3972 strain to chemotype IV (chemotype of O4). A structural analysis of LPS isolated from KX-V212 revealed that the inner core region of the LPS consisted of only one mole of 2-keto-3-deoxy-D-manno octonic acid, which carried a phosphate group at position C4 and the outer core at position C5. In passive hemolysis tests performed by using LPS as the antigen to sensitize sheep red blood cells (SRBC), and diagnostic antisera (O1 to O11) or anti-whole-cell rabbit antisera raised against O12, O13 and the six OUT strains, strong cross-reactivity was observed among LPS derived from the strains belonging to chemotype II (U-6443, W-90144, AD-7999 and O2). Strong cross-reactivity was also observed between X-3972 (chemotype IV) and O4 LPS. In contrast, LPS from two of the strains belonging to chemotype III (90A-6611 and KX-V212) did not react with any of the antisera raised against known O-serotypes. Cross-absorption tests showed that the O-antigens of U-6443, W-90144 and AD-7999 were identical to that of O2, and the O-antigen of X-3972 to that of O4. On the other hand, after the absorption of antisera raised against 90A-6611 and KX-V212 with O2 cells, the hemolytic activities against SRBC sensitized with homologous LPS were still retained at a high titer, whereas the hemolytic activities against SRBC sensitized with LPS from other O-serotype strains were completely eliminated. A cross-absorption test revealed that the O-antigens of these two strains were identical to each other. Thus, it was demonstrated that the O-serotype of OUT strains 90A-6611 and KX-V212 was not involved in the known O-serotypes; rather it represented a novel serotype which has not hitherto been reported. PMID- 10832966 TI - Oxalate-degrading Enterococcus faecalis. AB - An oxalate-degrading Enterococcus faecalis was isolated from human stools under anaerobic conditions. The bacteria required a poor nutritional environment and repeated subculturing to maintain their oxalate-degrading ability. The E. faecalis produced 3 proteins (65, 48, and 40 kDa) that were not produced by non oxalate-degrading E. faecalis as examined by SDS-PAGE. Antibodies against oxalyl coenzyme A decarboxylase (65 kDa) and formyl-coenzyme A transferase (48 kDa) obtained from Oxalobacter formigenes (an oxalate-degrading anaerobic bacterium in the human intestine) reacted with 2 of the proteins (65 and 48 kDa) from the E. faecalis as examined by Western blottings. This is the first report on the isolation of oxalate-degrading facultative anaerobic bacteria from humans. PMID- 10832967 TI - Acyclovir resistance in herpes simplex virus isolates from keratitis cases: an analysis from a developing country. AB - Seven herpes simplex type-1 (HSV-1) isolates from herpes simplex keratitis (HSK) cases clinically resistant to acyclovir (ACV) were analyzed for the mechanism of ACV resistance in them. The purpose of the study was to focus the attention of ophthalmologists on the frequency of occurrence of ACV resistance in HSK and to characterize such a phenomenon. We employed in-vitro plaque reduction assay, thymidine kinase assay, polymerase chain reaction, single-strand confirmation polymorphism analysis and sequencing to detect any mutation(s) in thymidine kinase gene in this analytical study. Four of the seven HSV-1 isolates proved ACV resistant by plaque reduction assay and three of them showed reduced thymidine kinase activity. Altered mobility pattern indicative of mutation within 335 base pair PCR product bracketing the suggested homopolymer mutational hotspot (7 Guanosine) was detected in 2 of these 3 isolates. DNA sequencing showed a deletion at nucleotide position 336 from the tk gene transcription start in both the isolates. This mutation has generated the first TGA stop codon 27 nucleotides downstream in the tk open reading frame. Our study also suggests the need of clinical/molecular surveillance of ACV resistance in HSV types in a given geographic location for better management of HSV infections. PMID- 10832968 TI - A simple screening method for detecting bindings between oligopeptides and HLA-DR molecules on filter papers: possible application for mapping of putative helper T cell epitopes on MSP1 of Plasmodium falciparum. AB - Binding capacities of synthetic peptides to HLA-DR molecules were tested on filter papers to identify putative helper T-cell epitopes on a malarial protein. The antigen tested was the merozoite surface glycoprotein 1 (MSP1) of Plasmodium falciparum, a vaccine candidate targeting the asexual erythrocytic stage. Bindings between synthetic oligopeptides and HLA-DR molecules were tested. Such bindings were not non-specific, and a known helper T-cell epitope peptide showed positive binding to the restricting HLA-DR molecule. By using this screening system, we observed the unequal distribution of HLA-DR-binding peptides in 10 out of 17 MSP1 blocks tested. Block #6 of MSP1 seemed to show the highest frequency in the positive binding; on the other hand, blocks #1 and #17, both of which were thought to be vaccine candidate regions, contained fewer HLA-DR binding peptides. This was not inconsistent with the results that block #17 was less stimulatory to peripheral T cells than block #6. The peptides with positive binding to HLA-DR showed actual epitope activities when we tested peptide-driven proliferation of human bulk T-cell lines, and association between the two parameters was statistically significant (P<0.001). For more detailed information for vaccine development, peptides with both IgG- and HLA-DR binding activities were mapped in block #17 of MSP1. Together with these results, we demonstrate that our simple screening system seems to provide essential information for vaccine development through uncovering locations of putative epitopes for human helper T cells. PMID- 10832969 TI - Intranasal sensitization of Japanese cedar pollen by the co-administration of low doses of cholera toxin but not its recombinant B subunit to mice. AB - We evaluated the effects of cholera toxin (CT) and the B subunit of cholera toxin (CTB) on the intranasal sensitization of Japanese cedar pollen (JCP) in mice. JCP suspended in phosphate-buffered saline was administered into the nostrils of mice in combination with varying doses of CT or recombinant CTB(r-CTB) once a week for 5 weeks. Antibody responses specific to sugi basic protein (SBP) were monitored by ELISA for seven weeks. The sensitization of JCP alone did not induce IgG1, IgG2b, IgG2a, IgE or IgA. In contrast, sensitization of JCP in combination with CT (JCP/CT) elicited the prominent production of SBP-specific IgG1 and low levels of IgG2b and IgG2a on Day 49. IgE production was detected only in the serum of mice which were treated with JCP/CT, and not under any other protocol. Using spleen cells from these mice, cytokine production was examined by ELISA in culture supernatants after they had been stimulated in vitro with major cedar pollen allergens, Cry j 1, Cry j 2 or SBP. Notable responses were an increase of IFN-gamma as well as IL-4 in JCP/CT-sensitized cells stimulated with Cry j 2, but not in those stimulated with Cry j 1. No significant differences were detected in IL-5 production among the experimental groups. Histopathological examination, however, showed that eosinophil infiltration was evident in the nasal mucosa of the JCP/CT-sensitized mice following challenge with JCP/CT, but weak with BSA/CT or CT alone. Thus, the immunological and histological analyses indicated that the co-administration of a low dose of CT in combination with JCP allows the induction of pollen-allergic states in mice. PMID- 10832970 TI - In vitro susceptibility of 7.5-kb common plasmid-free Chlamydia trachomatis strains. AB - Using a new plaque cloning technique, we obtained unique Chlamydia trachomatis strains which were confirmed to be free of the 7.5-kb common plasmid and glycogen in inclusions. The in vitro susceptibility of these strains to various chemotherapeutic agents was tested by comparison with their parent strains and clinical isolates possessing the common plasmid. No difference was detected for any of the agents tested, indicating that the 7.5-kb common plasmid is unrelated to the drug resistance of C. trachomatis. PMID- 10832971 TI - Release of membrane vesicles containing endotoxic lipopolysaccharide in Escherichia coli O157:H7 clinical isolates. AB - Membrane vesicles released by E. coli O157:H7 strains were investigated by immuno electron microscopy using anti-O157 antibody. Anti-O157 antibody enhanced the negative-staining of vesicles and we found numerous small vesicles clearly formed around bacterial cells. An immunogold-electron microscopic examination confirmed that lipopolysaccharide (LPS) including the O-side chain is present on the surface of the vesicles. Sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis of the purified vesicles showed that the vesicles contained LPS consisting of a lipid-A and an O polysaccharide. In addition, the endotoxic activity of the vesicle was confirmed by a limulus test. These results suggest that the vesicles may play an important role in the pathogenesis of Escherichia coli O157:H7. PMID- 10832972 TI - Cloning and sequencing of the gene encoding the candidate HtrA of Rickettsia typhi. AB - We screened a phage library of Rickettsia typhi with a polyclonal antiserum to clone genes which encode immunogenic proteins of R. typhi. Among several clones obtained, one clone codes for a 466-amino-acid protein similar to the heat-shock protein, HtrA. The deduced rickettsial HtrA contains a putative signal peptide sequence at the N-terminus, a serine protease-like domain, and two PDZ domains. The recombinant protein of rickettsial HtrA reacted with sera from patients with murine typhus and tsutsugamushi disease. We suggest that this gene and its recombinant protein would be valuable for the immunologic diagnosis of rickettsial diseases. PMID- 10832973 TI - Identification of a two-component signal transduction system involved in fimbriation of Porphyromonas gingivalis. AB - Porphyromonas gingivalis, a periodontopathogen, is an oral anaerobic gram negative bacterium with numerous fimbriae on the cell surface. Fimbriae have been considered to be an important virulence factor in this organism. We analyzed the genomic DNA of transposon-induced, fimbria-deficient mutants derived from ATCC 33277 and found that seven independent mutants had transposon insertions within the same restriction fragment. Cloning and sequencing of the disrupted region from one of the mutants revealed two adjacent open reading frames (ORFs) which seemed to encode a two-component signal transduction system. We also found that six of the mutants had insertions in a gene, fimS, a homologue of the genes encoding sensor kinase, and that the insertion in the remaining one disrupted the gene immediately downstream, fimR, a homologue of the response regulator genes in other bacteria. These findings suggest that this two-component regulatory system is involved in fimbriation of P. gingivalis. PMID- 10832974 TI - Autocrine interferon-beta stimulation augments nitric oxide production by mouse macrophage J774A.1 cells infected with herpes simplex virus type 1. AB - The pathogenic roles of nitric oxide (NO) in mouse models have been reported for herpes simplex virus type 1 (HSV-1)-induced pneumonia as well as endotoxin shock. We compared the mechanism of NO production induced by HSV-1 with that induced by lipopolysaccharide (LPS) using a mouse macrophage cell line, J774A.1. Both HSV-1 and LPS induced NO production as well as antiviral activity, which were attenuated by anti-interferon (IFN)-beta treatment. These results suggest that autocrine IFN-beta plays a role in NO release by J774A.1 cells stimulated with HSV-1 or LPS. PMID- 10832976 TI - Preventive effect of Ninjin-to (Ren-Shen-Tang), a Kampo (Japanese traditional) formulation, on spontaneous autoimmune diabetes in non-obese diabetic (NOD) mice. AB - We previously found that ingestion of an extract of Ninjin-to (NJT; Ren-Shen Tang) suppressed the development of autoimmune diabetes in C57BL/KsJ mice induced by multiple low doses of streptozotocin. To verify this effects on spontaneous autoimmune diabetes, the effects of NJT on NOD mice were investigated in the present study. NJT, provided in drinking water (0.25%, 450 mg/kg/day) from 6 weeks of age, significantly prevented the incidence of spontaneous diabetes in female NOD mice at 30 weeks of age (2/10) compared with that of the controls (7/10), with no effects on body growth or food intake. Even in non-diabetic mice, the blood glucose levels of the NOD controls gradually increased with age, while such increase in NJT-treated mice was significantly suppressed by preventing any deficiency of glucose tolerance. NJT also significantly suppressed the progression of insulitis, which causes insulin deficiency and diabetes. It is well known that NOD mice develop insulitis and diabetes because of their Th1 dominant autoimmune response. IFN-gamma production from splenic T lymphocytes stimulated with anti-CD3 monoclonal antibodies was increased, whereas IL-4 production was decreased in NOD controls compared to age- and sex-matched normal ICR mice. NJT-treatment reduced these deviations of cytokine production in NOD mice. These data all suggest that NJT can prevent spontaneous insulitis and diabetes by the modification of deviated cytokine production in NOD mice. PMID- 10832975 TI - Reduction of thymocyte numbers in transgenic mice expressing viral FLICE inhibitory protein in a Fas-independent manner. AB - A viral FLIP (FLICE/caspase-8-Inhibitory Protein), equine herpesvirus type 2 E8 protein, has been shown to inhibit Death receptor-induced apoptosis by suppressing the activation of FLICE/caspase-8. We generated transgenic mice specifically expressing E8 in thymocytes under the control of lck-proximal promoter. Although E8-expressing thymocytes were resistant to Fas-mediated apoptosis, the total number of thymocytes in 4-8-week-old E8 transgenic mice was more than 3-fold less than that in control littermates. This reduction was also observed in E8 transgenic mice with a Fas-/- background suggesting the reduction to be independent of Fas. The thymocytes of the transgenic mice, however, could similarly respond to CD3-mediated stimulation, indicating that the reduction of thymocyte numbers might be independent of T cell receptor complex-mediated stimulation. Thus, the Death receptor-mediated signaling pathway is too complex to be regarded as only an executor for apoptosis. PMID- 10832977 TI - Burkholderia uboniae sp. nov., L-arabinose-assimilating but different from Burkholderia thailandensis and Burkholderia vietnamiensis. AB - A polar multitrichous gram-negative motile rod, EY 3383, originally identified as Burkholderia thailandensis, revealed a DNA-DNA reassociation rate of 36.7%, under stringent conditions, with the type strain of B. thailandensis, despite the 16S rDNA homology value between two type strains being as high as 97.9%. The strain was clearly differentiated from the type strain of B. thailandensis by physiological, bio-chemical, and nutritional characteristics, without significant difference in cellular fatty acid and lipid composition. Based on the results of 16S rDNA sequence analysis, DNA-DNA hybridization and phenotypic characterization, Burkholderia uboniae sp. nov. is herein proposed. The type strain is NCTC 13147=EY 3383, isolated on 8 December 1989 from surface soil along the roadside in Ubon Ratchathani, Thailand. Major respiratory quinone is ubiquinone-8(Q8). G+C content of DNA is 69.71%. PMID- 10832978 TI - The role of osteopontin in the development of granulomatous lesions in lung. AB - Osteopontin (OPN) has been shown to be expressed by cells in granulomas of various origins, but whether it plays a functional role in granuloma formation is not known. Here we used a cardiomyopathic hamster (TO2) model, to test the hypothesis that OPN contributes functionally to granuloma development. We immunized cardiomyopathic and normal hamsters by subcutaneous injection of bovine serum albumin in complete Freund's adjuvant, and assessed various tissues for both OPN RNA expression and granuloma formation. Cardiomyopathic hamsters expressed OPN, and formed granulomatous lesions, in heart tissue in both immunized and untreated animals. In addition, immunization induced expression of OPN in lung and lymph nodes of cardiomyopathic (but not normal) hamsters, and also induced granuloma formation in these organs. To test whether OPN expression could play a functional role in inducing granulomas, we produced an adenoviral vector containing the murine OPN gene, and introduced this vector intratracheally into the lungs of normal hamsters. The OPN-containing vector, but not the control vector, induced pulmonary granuloma formation. These studies provided direct in vivo evidence that OPN can contribute functionally to the formation of granulomatous lesions, and suggest that OPN expression may be a common factor involved in formation of granulomas of various origin. PMID- 10832979 TI - Long-term results of correction of high myopia with an iris claw phakic intraocular lens. AB - PURPOSE: Anterior chamber phakic intraocular lenses (PIOLs) are one of the modalities used to correct high myopia. We report the long-term results of our prospective study on the Artisan 5-mm optic myopia lens. METHODS: We studied 67 eyes of 38 consecutive patients with preoperative myopia ranging from -5.38 to 28.00 D. All patients were operated by one surgeon. Mean follow-up was 35 months (24 months in 67 eyes and 36 months in 61 eyes). RESULTS: In 45 eyes (67.2%), postoperative residual refraction was within +/- 1.00 D of emmetropia. The mean refraction was stable statistically during the entire follow-up period. Mean best spectacle-corrected visual acuity improved from 20/40 to 20/32. Mean endothelial cell loss at 6 months was 5.5% (range, -52.4% to +9.3%), at 12 months, 7.21% (range, -53.2% to +20.1%), at 24 months, 9.1% (range -43.6% to +13.6%), and at 36 months, 10.9% (range, -43.0O% to +11.4%). The majority of eyes had an increase in best spectacle-corrected visual acuity; 5 eyes lost best spectacle-corrected visual acuity. We encountered no major complications. CONCLUSION: Implanting the Artisan 5-mm optic myopia lens in high myopic eyes resulted in a stable and accurate refractive outcome. The apparent progressive corneal endothelial cell loss remains a matter of concern. PMID- 10832980 TI - Holmium laser thermal keratoplasty for hyperopia and astigmatism after photorefractive keratectomy. AB - PURPOSE: To report results of holmium laser thermal keratoplasty used to treat induced hyperopia and induced, as well as pre-existing astigmatism after photorefractive keratectomy. METHODS: Sixteen eyes of 16 patients were included in this study. Contact holmium laser (Technomed Holmium 25) was used in 7 patients to correct hyperopia (8 spots at 8 or 9 mm) and in 9 patients to correct astigmatism (4 spots at 7, 8, or 9 mm). Follow-up evaluation was done after at least 6 months. The effectiveness, stability, and safety of the procedure were investigated. RESULTS: Spherical correction was ineffective (1.00 D or less) when applied at the 9-mm diameter treatment zone. Spherical correction applied at the 8-mm diameter treatment zone was ineffective in 1 eye. Three eyes achieved 1.00 to 2.00 D change, but 2 of these eyes showed an induced astigmatic change as well. Correction of astigmatism at the 7-mm diameter treatment zone resulted in a 0 to 4.00 D cylinder component change. Treatment at the 8-mm diameter treatment zone showed a 0 to 1.50 D effect and at the 9-mm treatment zone, 0.25 to 1.50 D. All eyes that achieved significant improvement (1.00 D or more change in cylinder component) showed significant overcorrection in the first postoperative phase. There were no sight threatening complications. CONCLUSION: Holmium laser thermal keratoplasty can be useful for the treatment of overcorrection and induced as well as pre-existing astigmatism after photorefractive keratectomy. However, predictability is low and astigmatism can be induced with the attempted spherical correction. PMID- 10832981 TI - Corneal endothelial cell damage after experimental diode laser thermal keratoplasty. AB - PURPOSE: To evaluate the safety of diode laser thermal keratoplasty (LTK) with respect to corneal endothelial cell damage. METHODS: In an in vitro animal model system, porcine eyes were irradiated with a continuously emitting laser diode at wavelengths (lambda) of 1.85 or 1.87 microm, corresponding to an absorption coefficient (micro(a)) of 1.1 or 2.0 mm(-1). Different irradiation and application parameters were tested serially. To determine the temperature threshold for endothelial damage, corneal buttons were analyzed separately in a waterbath experiment. The endothelial damage was assessed after trypan blue and alizarin red supravital staining under light microscopy. RESULTS: The thresholds for the 50% probability of thermal damage (ED50) were determined at corneal temperatures of 65 degrees C for a 10-second water-bath immersion, and 59 degrees C for 60 seconds. Coagulations that reached the deeper stromal layers revealed severe endothelial cellular alterations and areas of exposed Descemet's membrane. The thermally induced changes were dependent on laser power and the absorption coefficient (wavelength). Mean diameter of total endothelial cell damage was 245 +/- 154 microm (range, 0 to 594 microm) for an absorption coefficient of 1.1 mm( 1). The maximal lateral extent of endothelial cell damage induced by the laser exposure was 594 microm in diameter. Increasing the absorption coefficient decreased the penetration depth of the laser irradiation, creating a greater temperature rise within the corneal stroma and significantly less endothelial damage (P < .01), when the same laser power was applied. The calculated total area of damage for the paracentral human corneal endothelium ranged from 1.8% to 13.6%. CONCLUSION: Data obtained in this in vitro study were transferred to an endothelial cell damage nomogram, demonstrating that appropriate parameter improvements can minimize the adverse effects to the corneal endothelium. However, model adjustment to the human cornea indicates the potential for endothelial cell damage after diode laser thermal keratoplasty, and should be considered when performing this elective procedure. PMID- 10832982 TI - A model to explain the difference between changes in refraction and central ocular surface power after laser in situ keratomileusis. AB - PURPOSE: After refractive surgery, changes in central ocular surface power are usually less than actual changes in refraction. The aim of this paper was to examine the theoretical changes in the radius of the epithelial-stromal interface and corneal stromal refractive index which could take place to account for some of the empirical findings. METHODS: The parameters of an aspheric human eye model featuring gradient index optics was modified using data from a clinical study evaluating effects of laser in situ keratomileusis (LASIK) for correction of moderate to high myopia. Computations were performed to determine: 1) theoretical postoperative radius of the epithelial-stromal interface and hence distribution of epithelial thickness, and 2) refractive index of the stroma when the epithelium was of fixed uniform thickness. RESULTS: Within the central 2-mm diameter refractive zone of the cornea after LASIK for myopia, either of two factors could account for the difference between changes in central ocular surface power and the actual change in ocular refraction: 1) a steepening of the epithelial-stromal interface resulting from a 400% centrifugal increase in epithelial thickness, or 2) a reduction in stromal refractive index from an average of 1.376 to 1.364. CONCLUSION: The difference between the observed changes in refraction and central ocular surface power could be explained by reduction in stromal refractive index and this could be secondary to a 6.5% increase in water content of the stroma during the postoperative period. The epithelial-stromal interface after LASIK is unlikely to steepen by the extent predicted by our model. This surface is not the source of the difference between the change in refraction and change in central ocular surface power. PMID- 10832983 TI - Relationship between anterior chamber depth, refractive state, corneal diameter, and axial length. AB - PURPOSE: Phakic intraocular lenses are being used increasingly to correct refractive errors. We studied the relationship between anterior chamber depth, refractive state of the eye, spherical equivalent refraction, axial length of the globe, corneal diameter, and keratometry. METHODS: Two hundred eleven eyes of 211 patients were enrolled. All eyes underwent the same protocol with a complete ocular examination that included slit-lamp microscopy, intraocular pressure, objective and subjective refraction, calculation of the spherical equivalent refraction, corneal pachymetry, anterior chamber depth, axial length of the globe, and keratometry. All results were analyzed statistically using SPSS statistics software. Correlations between different parameters were studied using the Pearson correlation test. RESULTS: The anterior chamber depth was found to correlate significantly with both the average corneal diameter and the axial length of the globe (0.744, 0.531, P < .01) and was also found to correlate through an inverse relation with both age and spherical equivalent refraction ( 0.391, -0.623, P < .01). Corneal thickness and keratometric power did not correlate with the anterior chamber depth. CONCLUSION: Most parameters (axial length, corneal diameter, spherical equivalent refraction, patient age) affected anterior chamber depth and should be considered carefully when planning refractive procedures that employ phakic intraocular lenses. PMID- 10832984 TI - Corneal morphology in vitro after superficial keratectomy with q-switched Er:YSGG and free-running Er:YAG lasers. AB - PURPOSE: Examination of morphology in corneal ablation induced by a q-switched Er:YSGG (2.79 microm) laser and a free-running Er:YAG laser (2.94 microm). METHODS: Defined ablation of 6-mm diameter and 15, 30, 55, 90, and 120-microm depth was performed on freshly enucleated swine eyes. Er:YSGG laser parameters: fluence 1.6 J/cm2, frequency 6 Hz, spot-size 465 microm FWHM, scanning-mode, pulse number 1520 to 6210. Er:YAG laser parameters: fluence 3 J/cm2, frequency 1.5 Hz, spotsize 6 mm, wide area ablation, pulse number 2 to 13. Corneal morphology was analyzed by gross photography, histology, scanning electron microscopy, and scanning nearfield acoustic microscopy. RESULTS: Histology showed thermal damage of 5 to 15 microm in depth caused by the Er:YSGG laser in comparison with 10 to 20 microm by the Er:YAG laser. Average roughness of the ablated surface measured with scanning nearfield acoustic microscopy was 20 to 40 microm for the Er:YSGG laser and 5 to 15 microm for the Er:YAG laser. These data confirm the subjective impression of images created by scanning electron microscopy and gross photography. CONCLUSIONS: Although the Er:YAG laser system appeared to demonstrate a smoother corneal surface than the q-switched Er:YSGG laser, the thermal damage in either case poses a potential limitation for clinical use in lamellar refractive surgery. PMID- 10832985 TI - Intraocular lens power calculation in eyes after corneal refractive surgery. AB - PURPOSE: The purpose of this review article is to discuss the major reasons for postoperative hyperopia after cataract surgery following radial keratotomy (RK) and photorefractive keratectomy (PRK) and to illustrate potential methods for improvement of intraocular lens (IOL) power prediction after keratorefractive surgery based on exemplary model calculations. METHODS: We previously performed model calculations in eyes after PRK for myopia (-1.50 to -8.00 D, mean -5.40 +/- 1.90 D) using keratometry readings as measured by the Zeiss keratometer and the TMS-1 topography unit and as calculated using the "clinical history method" (spherical equivalent refraction change) and change in anterior surface keratometry readings. RESULTS: We found that after PRK, mean measured keratometry readings were significantly greater than respective calculated values considering the preoperative to postoperative change of anterior corneal surface (P < .001), which itself was significantly greater than calculated keratometry readings considering the preoperative to postoperative change of spherical equivalent refraction (P < .001). IOL power underestimation correlated significantly with the difference between preoperative and postoperative spherical equivalent refraction (P = .001). CONCLUSIONS: For correct assessment of keratometric readings to be entered into more than one modern third-generation IOL power calculation formula (but not a regression formula), the clinical history method should be applied whenever refraction and keratometric diopters before the keratorefractive procedure are available to the cataract surgeon. If preoperative keratometric diopters and refraction are not known, average central power on the postoperative videokeratograph may be used after RK, but refined calculation of keratometric diopters from radius of anterior and posterior corneal surface should be used after PRK and/or LASIK. PMID- 10832986 TI - Decreased tear secretion after laser in situ keratomileusis for high myopia. AB - PURPOSE: To investigate the changes in tear secretion and tear film stability after excimer laser in situ keratomileusis (LASIK) for high myopia. METHODS: One eye of 28 patients underwent unilateral LASIK for the correction of myopia with spherical equivalent refractions ranging from -6.37 to -18.25 D (mean, -11.98 +/- 3.45 D). Schirmer tear test values without anesthesia at 5 minutes and tear break up time values were measured 4 weeks after LASIK. Unoperated contralateral eyes were used as a control for each patient. The Schirmer tear test and tear break-up time values were analyzed statistically using the independent t-test. RESULTS: The mean Schirmer tear test value was 16.17 +/- 2.50 mm in the operated eyes and 21.07 +/- 7.03 mm in the unoperated (control) eyes of the same patients (P = .001). The mean tear break-up time value was 21.0 +/- 3.55 seconds in the operated eyes and 21.27 +/- 6.79 seconds in the control eyes (P = .85). CONCLUSION: Tear secretion decreased following LASIK probably due to decrease in corneal sensitivity, but tear film stability appeared unaltered. PMID- 10832987 TI - Intrastromal corneal ring segments in a patient with previous laser in situ keratomileusis. AB - PURPOSE: Intrastromal corneal ring segments (ICRS; Intacs) were inserted in a patient with residual myopia of -3.375 D (spherical equivalent) 10 months after laser in situ keratomileusis (LASIK). METHODS: A standard intrastromal corneal ring segment implantation technique was used with the addition of intraoperative ultrasonic pachymetry in 4 quadrants at the 7-mm zone to insure adequate stromal thickness for segment insertion. RESULTS: Four months after ICRS surgery and 14 months after LASIK, the patient had uncorrected visual acuity of 20/20 and a cycloplegic refraction of plano -1.00 x 23 degrees. CONCLUSION: Implantation of intrastromal corneal ring segments in an eye with previous LASIK resulted in additional corneal flattening with a decrease in residual myopia and improved uncorrected visual acuity. PMID- 10832988 TI - Keratectasia induced by laser in situ keratomileusis in keratoconus. AB - PURPOSE: Corneal thinning disorders weaken the mechanical strength of affected corneas, suggesting that photorefractive procedures may be contraindicated in keratoconus. Few cases have been reported to confirm this hypothesis. METHODS: A 45-year-old man had two laser in situ keratomileusis (LASIK) procedures and one photorefractive keratectomy (PRK) performed on his left eye, and three LASIK procedures on his right eye. After these surgeries, a dramatic corneal ectasia and grade III haze occurred in both eyes, with a clinical diagnosis of keratoconus. The changes in his corneas were followed with videokeratography and slit-lamp microscopy. RESULTS: Preoperative videokeratograph of both eyes suggested keratoconus. After multiple refractive procedures, the best spectacle corrected visual acuity was as low as 20/1200 bilaterally. Both eyes displayed dramatic corneal protrusion with corneal scarring. CONCLUSIONS: This case emphasizes the need for preoperative corneal thickness measurement and detailed analysis of videokeratographs. Thinning corneal disorders such as keratoconus, keratoconus suspects, or pellucid marginal degeneration are a contraindication for excimer laser ablative refractive procedures. PMID- 10832989 TI - Premacular subhyaloid hemorrhage following laser in situ keratomileusis. AB - PURPOSE: To report a case of premacular subhyaloid hemorrhage following laser in situ keratomileusis (LASIK). METHODS: Case report. RESULTS: The subhyaloid hemorrhage did not resolve over 1 month of observation, necessitating Nd:YAG posterior hyaloidotomy. CONCLUSION: The LASIK procedure can be associated with postoperative subhyaloid hemorrhage, presumably from rapid release of the microkeratome vacuum pressure. PMID- 10832990 TI - Partial dislocation of laser in situ keratomileusis flap by air bag injury. AB - PURPOSE: A patient developed significant corneal complications from air bag deployment, 17 months after laser in situ keratomileusis (LASIK). METHODS: Case report, slit-lamp microscopy, and review of the medical literature. RESULTS: A 37 year-old woman underwent bilateral LASIK with resultant 20/20 uncorrected visual acuity. Seventeen months later, she sustained facial and ocular injuries from air bag deployment during a motor vehicle accident. Examination revealed bilateral corneal abrasions, partial dislocation of the right corneal LASIK flap, and a hyphema in the right eye. The LASIK flap was realigned, but recovery was complicated by a slowly healing epithelial defect and flap edema. One month following the injury, epithelial ingrowth beneath the LASIK flap was noted. Surgical elevation of the flap and removal of the epithelial ingrowth was performed. Eight months later, epithelial ingrowth was absent and the visual acuity was 20/40. Residual irregular astigmatism necessitated rigid gas permeable contact lens fitting to achieve 20/20 visual acuity. CONCLUSIONS: Air bags may cause significant ocular trauma. The wound healing response of LASIK allows corneal flap separation from its stromal bed for an indeterminate time after surgery. Discussion of the possible risk of corneal trauma as part of informed consent prior to LASIK may be appropriate. PMID- 10832991 TI - Induced astigmatism following laser in situ keratomileusis for myopia with a free cap. PMID- 10832992 TI - Flap complication rate for two microkeratomes. PMID- 10832993 TI - Evaluation of corneal wound healing after excimer laser keratectomy. PMID- 10832994 TI - Results of Internet poll on outcome of LASIK in keloid formers. PMID- 10832995 TI - Porcine eyes as an experimental model for wound healing. PMID- 10832996 TI - Your patients are on drugs. PMID- 10832997 TI - Regenerate bone formation and remodeling during mandibular osteodistraction. AB - The purpose of this study was to evaluate the newly formed bone during the consolidation period of mandibular osteodistraction using quantitative histology. Seventeen skeletally mature conditioned male beagle dogs underwent 10 mm of bilateral mandibular lengthening. After distraction, the regenerates were allowed to consolidate for 0, 2, 4, 6, or 8 weeks, at which time the animals were sacrificed and tissues harvested for standard histologic and histomorphometric analyses. Mineralization began at the host bone margins by the end of the distraction period, followed by a progressive increase in trabecular bone, with a concomitant decrease in the amount of fibrous tissue. Between 4 to 6 weeks of consolidation, 3 types of relatively mature distraction regenerates were evident. The mineral apposition rate gradually increased from the end of distraction to the fourth week of consolidation, at which time it remained constant until sometime before the eighth week, when it tapered off slightly as remodeling increased. PMID- 10832998 TI - Long-term stability of surgical open-bite correction by Le Fort I osteotomy. AB - Skeletal changes greater than those observed in untreated adults have been noted beyond 1 year post-surgery in adult patients who had surgical correction of a long face deformity. The stability of skeletal landmarks and dental relationships from 1 to >3 years post-surgery was examined in 28 patients who had undergone surgery of the maxilla only, and in 26 patients who had undergone 2-jaw surgery to correct >2 mm anterior open bite. Although the average changes in almost all landmark positions and skeletal dimensions were less than 1 mm, point B moved down >2 mm and face height increased >2 mm in one-third of the maxilla-only group and in 40% of the 2-jaw group (>4 mm in 10% and 22% respectively). Overbite decreased 2-4 mm in only 7% of the maxilla-only and 12% of the 2 groups, with no changes >4 mm, because in three-fourths of the patients with an increase in anterior face height, further eruption of the incisors maintained the overbite relationship. In the maxilla-only group, mandibular length (Co-Pg) showed >2 mm long-term change in 45% of the patients, two-thirds of whom showed an increase rather than a decrease in length. In the 2-jaw group, no patients showed a decrease in Co-Pg length and one-third had an increase. For both groups, changes in overjet were smaller and less frequent than changes in mandibular length. PMID- 10832999 TI - Orthodontic tooth movement in the prednisolone-treated rat. AB - Adverse effects of corticosteroids on bone metabolism raise concerns as to whether steroid treatment may influence orthodontic movement. This study examined the effect of prednisolone on orthodontic movement using an established rat model. The corticosteroid treated group (N = 6) was administered prednisolone (1 mg/kg) daily, for a 12-day induction period; the control group (N = 6) received equivalent volumes of saline. On day 12, an orthodontic appliance was placed which exerted 30 g of mesial force to the maxillary first molar. Animals were sacrificed on day 24 and tooth movement was measured. Sagittal sections of the molars were stained with haematoxylin and eosin, and for tartrate-resistant acid phosphatase (TRAP) activity. While there were no significant differences in the magnitude of tooth movement between the 2 groups, steroid-treated rats displayed significantly less root resorption on the compression side and fewer TRAP positive cells within the PDL space on the same side. This suggests steroid treatment suppressed clastic activity. PMID- 10833000 TI - Tooth-size reduction associated with occurrence of palatal displacement of canines. AB - This study investigates mesiodistal crown size of the maxillary and mandibular incisors of patients with palatally displaced canines (PDC). Pretreatment dental casts of orthodontic patients with PDC of 1 or both maxillary canines (N = 31; M10:F21) were collected. This PDC sample was matched according to age and sex with pretreatment dental casts from unaffected orthodontic patients. For the PDC and matched control samples, maximum mesiodistal crown diameters were recorded for the 4 incisors on the left side only. The results showed that, on average, the mesiodistal crown diameters for the maxillary and mandibular incisors measured smaller in the PDC sample than in the control sample. These findings of statistically significant tooth-size reductions associated with PDC occurrence indicate a generalized pattern of reduced tooth size as a characteristic associated with the PDC anomaly. Further, the presence of generalized tooth-size reduction in cases with palatally displaced canines helps explain why most orthodontic treatment plans for PDC patients are of the nonextraction type. PMID- 10833001 TI - Nonsurgical rapid maxillary expansion in adults: report on 47 cases using the Haas expander. AB - Rapid maxillary expansion (RME) in the adult is thought to be an unreliable procedure with several adverse side effects and, consequently, surgically assisted RME is considered the preferred procedure. The purpose of this paper is to study the efficacy of nonsurgical RME, and to determine the incidence of complications such as relapse of the expansion, pain and tissue swelling, tipping of the molars, opening rotation of the mandible and gingival recession. Rapid maxillary expansion using a Haas expander was examined in 47 adults and 47 children. A control group of 52 adult orthodontic patients who did not require RME was also studied. Students' t-test, and the analysis of variance followed by the Scheffe test were used to determine if there were significant differences among time periods and among the 3 study groups. The mean transarch width increase was similar in adults and children who had RME; 4.6 +/- 2.8 compared to 5.7 +/- 2.4 mm for the molars and 5.5 +/- 2.4 compared to 5.7 +/- 2.5 mm for the second premolars. In the adults, transarch expansion and the correction of the posterior crossbites were stable following discontinuance of retainers (mean 5.9 years). If the expander was properly fabricated, and turned no more than once a day, the procedure was well-tolerated. Rapid maxillary expansion in adults flared the molars buccally only 3 degrees per side. The mandibular plane and lower facial height were unchanged. The adults achieved 18% of their transmolar expansion at the height of the palate and the remainder with buccal displacement of the alveolus. The children achieved 56% of their expansion by an increase at the height of the palate with the remainder due to displacement of the alveolus. There was some buccal attachment loss (0.6 +/- 0.5 mm) seen in the female subjects associated with RME, but the extent was clinically acceptable. This resulted in significantly longer clinical crowns, but rarely caused exposure of buccal root cementum. Complications were infrequently observed or of minimal consequence. The results indicate that nonsurgical RME in adults is a clinically successful and safe method for correcting transverse maxillary arch deficiency. PMID- 10833002 TI - Dentoalveolar compensation in negative overjet cases. AB - The purpose of this study was to investigate dentoalveolar compensation in negative overjet cases. Eighty-eight adult females with either skeletal Class I or skeletal Class III jaw relationships were examined. Of the total, 44 cases showed anterior crossbite and the remaining 44 cases had normal incisor relationships. Four cephalometric parameters were measured: the sagittal jaw relationship, maxillary and mandibular incisor inclination, and the occlusal plane angulation. In the negative overjet cases, correlation analysis was performed between the skeletal and dental measurements. Stepwise discriminate analysis was carried out to separate the negative and normal overjet cases. Compensatory changes for sagittal jaw discrepancies in the negative overjet cases were statistically confirmed for both incisor inclination and occlusal plane angulation. However, the compensatory effects were weaker than in the normal overjet cases. The discriminate analysis successfully separated the normal and negative overjet cases, suggesting that negative overjet results from insufficient dentoalveolar compensation for variations in the sagittal jaw relationships. PMID- 10833003 TI - Comparisons of two approaches for removing excess adhesive during the bonding procedure. AB - The purpose of this study was to compare the effects on shear bond strength of removing excess adhesive from around the bracket base at 2 time periods: (1) immediately after placing the bracket on the tooth, and (2) after subjecting the adhesive to 5 seconds of light curing to initially secure the bracket in its proper position. The debonding forces were evaluated at 2 times; within half an hour after bonding and after storing for 24 hours in water at 37 degrees qC. These comparisons will help determine the most advantageous time for the clinician to remove excess adhesive from around the brackets during the bonding process. The teeth were randomly divided into 4 groups according to: (a) the time of removal of the excess adhesive from around the bracket base namely; immediately after placing the bracket or after 5 seconds of light cure and (b) the time of debonding the brackets, namely within half an hour or after 24 hours. Shear bond strength was measured using a Zwick test machine and calculated in Megapascals. The results of the analysis of variance (F = 35.05) comparing the 4 experimental groups indicated the presence of significant differences between all 4 groups (P = .0001). In general, the shear bond strengths were significantly larger for the 2 groups debonded after 24 hours, whether they were light cured for a total of 40 seconds (X = 8.8 +/- 3.6 MPa) or 45 seconds (X = 6.9 +/- 3.4 MPa). On the other hand, the shear bond strengths was significantly lower in the 2 groups debonded within half an hour from their initial bonding, whether light cured for 40 seconds (X = 0.4 +/- 1.0 MPa) or 45 seconds (X = 3.4 +/- 2.7 MPa). In conclusion, the additional 5 seconds of light cure significantly increased the initial shear bond strength. On the other hand, removing excess adhesive after 5 seconds of light cure significantly decreased the shear bond strength at 24 hours. PMID- 10833004 TI - A formula to determine the amount of retraction of mandibular canines. AB - Moderate to severe protrusion of anterior teeth often necessitates the extraction of 4 premolars to retract anterior teeth into their space. We present a formula that determines how far the mandibular canines should be retracted to provide adequate space for correction of lower incisor protrusion. To test the accuracy of the formula, 23 mandibular setups were made and the results of the formula were compared with those of the setups. Paired t-tests at a confidence level of 95% did not show any statistically significant differences between the results. PMID- 10833005 TI - Validity of identification of gonion and antegonion in frontal cephalograms. AB - This study was designed to develop a method of transferring gonion from lateral to frontal cephalograms, and to use this method as gold standard when evaluating observer performance in identifying gonion in frontal cephalograms. Observer ability to identify antegonion was also evaluated. There was a range of 28 mm in the observers' identification of gonion and a statistically significant deviation from gold standard. The factors "observer" and "cephalogram," regarded as random effects in an ANOVA analysis, and their interaction, each influenced the result, P < .001. The deviation from the mean of all observations for antegonion ranged 8 mm with "cephalogram" having a statistically significant influence. The results suggest that neither gonion nor antegonion can be routinely used as valid landmarks in frontal cephalograms. Gonion can, however, be used if first identified in a lateral cephalogram and transferred to a paired frontal cephalogram aided by radiographic indicators combined with a bilateral scrutiny of projection geometry in different planes through gonion and indicator. PMID- 10833006 TI - Treatment of mandibular asymmetry by distraction osteogenesis and orthodontics: a report of four cases. AB - Distraction osteogenesis devices followed by hybrid functional appliance therapy and fixed orthodontic appliance therapy were used to correct a variety of maxillofacial skeletal and dental deformities in 4 patients. The patients underwent procedures to lengthen the mandibular ramus and body. Transcutaneous pins were used to activate the distraction devices. After achieving the desired skeletal position, the distraction devices were maintained for 2 to 3 months to allow ossification. Following distraction, functional orthodontic appliance therapy was initiated during growth to correct the cant of the occlusal plane by extrusion of teeth on the affected side for improved facial symmetry. Fixed orthodontic therapy was used for final occlusal adjustments. All patients achieved lengthening of their jaws and substantial improvement in facial symmetry and occlusion. The follow-up period for this group of 4 patients ranged from 18 to 36 months. PMID- 10833007 TI - Secondary bone graft and eruption of the permanent canine in patients with alveolar clefts: literature review and case report. AB - This paper emphasizes the important role that secondary bone grafting plays on the treatment of patients with alveolar clefts. The authors present a literature review and, based on panoramic radiographs, retrospectively and longitudinally analyze the behavior of permanent canines after completion of secondary bone grafting in 50 patients at the Hospital for Rehabilitation of Craniofacial Anomalies, University of Sao Paulo, Brazil. Twelve patients with unilateral cleft lip and alveolus and 38 patients with complete unilateral cleft lip and palate (n = 50) had undergone bone grafts to repair their residual alveolar clefts before the eruption of their permanent canines. These patients were observed over an average period of 3 years. In 94% of the sample (47 patients), the permanent canines presented intra-alveolar movement toward the oral cavity. In 72% of those 47 patients (36 patients), the permanent canines spontaneously erupted through the grafted area. In 6% of the 47 patients (3 patients), the permanent canines required orthodontic traction. PMID- 10833008 TI - Phylogenetic analysis of Trypanosomatina (Protozoa: Kinetoplastida) based on minicircle conserved regions. AB - Phylogenetic relationships within the suborder Trypanosomatina were inferred from the kinetoplast DNA minicircle conserved region sequences. Trees built using distance-matrix (Neighbor-Joining) and maximum parsimony methods showed that the minicircle conserved regions (CRs) provide a sensitive and specific molecular marker suitable for phylogenetic analyses of subspecies and strains of trypanosomatid flagellates, as testified by the subdivision of the genus Leishmania into the subgenera Leishmania. Viannia and Sauroleishmania. However, since Phytomonas and monogenetic parasites of insects represent the earliest diverging groups, the CRs do not seem to be useful for inference of relationships among major lineages of the order Kinetoplastida. PMID- 10833009 TI - Preference of female rats for the odours of non-parasitised males: the smell of good genes? AB - Many animals obtain reliable information about potential mates, including whether they are parasitised or not, mostly from olfactory cues in urine. Previous experiments with rodents have shown that females can detect parasites in males that are potentially transmissible during copulation, so that females can directly avoid infection by discriminating against parasitised males. Here, using choice tests, we examine whether female rats can distinguish males infected with the tapeworm Hymenolepis diminuta Rudolphi, 1819, a parasite with a complex life cycle and thus not directly transmissible among rats. Female rats tended to spend more time investigating the urine of non-parasitised males than that of parasitised males. The magnitude of the parasite burden in the infected males had no effect on the females' preference for the non-parasitised males. We also found that parasitised males had lover testosterone levels in their blood than non parasitised males. These results suggest that females use cues in male urine reflecting either the presence of the parasite and/or lower testosterone levels to avoid parasitised males and possibly secure resistance genes for their offspring. PMID- 10833010 TI - Neopolystoma liewi sp. n. (Monogenea: Polystomatidae) from the eye of the Malayan box turtle (Cuora amboinensis). AB - Neopolystoma liewi sp. n. is described from the conjunctival cavity of the Malayan box turtle Cuora amboinensis (Daudin, 1802), in Peninsular Malaysia. This is the first record of Neopolystoma in Malaysia and the fourth polystomatid species described from C. amboinensis. Of the 27 Malayan box turtles examined, 8 were found to be infected. A maximum of 2 parasites per eye and 4 individuals per host was recorded. N. liewi sp. n. differs from all other members of the genus by possessing few and short genital spines and small marginal hooks. The oncomiracidium has 64 ciliated cells arranged symmetrically about the sagittal axis. PMID- 10833011 TI - Comparative infectivity of Fasciola hepatica metacercariae from isolates of the main and secondary reservoir animal host species in the Bolivian Altiplano high human endemic region. AB - Fascioliasis due to Fasciola hepatica (Linnaeus, 1758) is an endemic disease on the Northern Bolivian Altiplano, where human prevalences and intensities are the highest known, sheep and cattle are the main reservoir hosts, and pigs and donkeys the secondary ones. Investigations were carried out to study the viability of metacercariae experimentally obtained from eggs shed by naturally infected Altiplanic sheep, cattle, pigs and donkeys. A total of 157 Wistar rats were infected with doses of 5, 10, 20 and 150 metacercariae. Metacercariae aged for different number of weeks were used to analyse the influence of age on their viability. The number of worms successfully developed in each rat was established by dissection. Results obtained show that metacercarial infectivity is dependent upon storage time, being lower when metacercariae are older. The maximum longevity is 31 weeks using doses of 20 metacercariae per rat and 48 weeks with 150 metacercariae per rat, although in the latter case only a very low percentage of worms is recovered. Age-related infectivity of metacercariae from Altiplanic F. hepatica does not significantly differ from that of the liver fluke in lowlands of other countries. Concerning the influence of the isolate according to host species, results indicate that metacercarial viabilities of pig and donkey isolates are similar to the viabilities of metacercariae of sheep and cattle isolates. Thus, pig and donkey have a high transmission potential capacity concerning this aspect. This fact is of great importance for the control of human and animal fascioliasis in this highly endemic zone. PMID- 10833012 TI - Morphology of Bertiella studeri (Blanchard, 1891) sensu Stunkard (1940) (Cestoda: Anoplocephalidae) of human origin and a proposal of criteria for the specific diagnosis of bertiellosis. AB - Human material of an African specimen of Bertiella studeri (Blanchard, 1891), a typical intestinal cestode of monkeys, is described. Mature, postmature and gravid proglottides, and eggs, previously inadequately figured, are illustrated and photographed. The description of the species agrees with that provided by Stunkard (1940). A comparative study with other descriptions of the species is made in an attempt to clarify previous findings. The morphological differences reported in various earlier descriptions of the species suggest that B. studeri should be regarded as a "B. studeri species complex" Improvements are required in the descriptions of new future findings in order to clarify the specific diagnosis of human bertiellosis. Evidence suggests that a generalised diagnosis exclusively based on egg size and geographical distribution is insufficient to differentiate B. studeiri and Bertiella mucronata (Meyner, 1895), or additional species may be affecting humans. PMID- 10833013 TI - Ultrastructure of the scolex, rhyncheal system and bothridial pits of Otobothrium mugilis (Cestoda: Trypanorhyncha). AB - The ultrastructure of the scolex tegument, bothridial pits (=ciliated pits) and rhyncheal system of Otobothrium mugilis Hiscock, 1954 is described from plerocerci collected from the teleosts Arius graeffei Kner et Steindachner and Mugil cephalus Linnaeus. Scanning electron microscopy revealed that filamentous microtriches with shortened caps are abundant across the entire surface of the tegument. Palmate microtriches are dominant on the bothridia and their margins. The surfaces of bothridial pits were covered with large bifid microtriches. The bothridial pits are strongly muscularised invaginations of the tegument. Nervous tissues were not observed within the pits and it is probable that these structures function as accessory attachment structures. The wall of each tentacle sheath consists of one to three bands of fibrils, lined internally by a thin cytoplasmic layer. The tentacular walls are cellular, containing myofilaments. The fibrils of the tentacular walls are arranged into discrete blocks of parallel fibrils and appear to be intracellular. Tentacular walls are lined externally by a modified membrane with an external glycocalyx. Tentacular hooks are solid, bound externally by a membrane. The body of the hook contains numerous longitudinal canaliculi and an electron-opaque medulla lies at the centre of the hook. PMID- 10833014 TI - The effect of condensed tannins from seven herbages on Trichostrongylus colubriformis larval migration in vitro. AB - The effects of condensed tannins (CT) extracted from seven forages on the motility of the economically important nematode, Trichostrongylus colubriformis (Giles, 1892), were evaluated by using a larval migration inhibition (LMI) assay. The assay involved incubation of third stage (L3) exsheathed T. colubriformis larvae with CT extracted from Lotus pedunculatus, Lotus corniculatus, sulla (Hedysarum coronarium), sainfoin (Onobrychis viciifolia), Dorycnium rectum, Dorycnium pentaphyllum and dock (Rumex obtusifolius) and measurement of larval migration through nylon mesh with a 20 microm pore size. At 100 microg ml(-1), CT from L. pedunculatus, L. corniculatus, sulla, sainfoin, D. rectum, D. pentaphyllum and dock inhibited 20%, 10%, 15%, 25%, 28%, 32% and 27% of the larvae, respectively from passing through the sieves compared to controls (no CT added). At 1000 microg CT ml(-1), CT purified from D. pentaphyllum had the highest inhibitory activity (63%) against 1-month old larvae followed by sainfoin (59%), L. pedunculatus (57%), D. rectum (53%), dock (50%), sulla (40%) and L. corniculatus (37%). Seven-month old larvae were more sensitive to the action of CT than 1-month old larvae (P < 0.001). Addition of 2 microg polyethylene glycol ([PEG] per microg CT; to remove the effect of CT) eliminated 81-93% of the CT activity (P < 0.001) compared to incubations without PEG. The impact of CT on larval migration suggests a possible role for these plants in ruminant diets as a means to reduce dependence upon proprietary anthelmintics. PMID- 10833015 TI - Bacillary band ultrastructure of the fish parasite Capillaria pterophylli (Nematoda: Capillariidae). AB - In Capillaria pterophylli Heinze, 1933, two lateral bacillary bands extend along the whole body in female and male worms. A ventral bacillary band is present in females only. The bacillary bands consist of glandular and non-glandular cells, in the region between the nerve ring and the end of the stichosome, ciliated sense receptors in tight connection with gland are present. PMID- 10833016 TI - A study of proteases throughout the life cycle of Trichinella spiralis. AB - In the present report we study the proteolytic activity of the excretion secretion and crude extracts of different stages of Trichinella spiralis (Owen, 1835) Railliet, 1895, (muscle-stage larvae, adult worms before and after mating, and newborn larvae) using natural substrates (structural and hematic mammalian proteins). The analysis of the results allow us to set up a certain stage specificity, as well as an important relationship between the protease patterns throughout the parasite life cycle and how the parasite may overcome both mechanical and humoral barriers within the host. Muscle-stage larvae present a great activity against structural proteins (collagen), while newborn larvae and adult worms degrade principally hematic proteins (hemoglobin, fibrinogen and immunoglobulin G). PMID- 10833017 TI - Analysis of intestinal intraepithelial lymphocyte populations in experimental Trichinella spiralis infection of mice. AB - The potential role of intestinal intraepithelial lymphocytes (i-IELs) in the generation of host protective immunity after helminth infection was investigated using the Trichinella spiralis (Owen, 1835)/mouse model. In this study we found a significant rise of TCRgamma(delta)+ i-IELs (P < 0.001) concurrent with the jejunal goblet cells (GC) hyperplasia in T. spiralis-infected C57BL mice on day 4 p.i. However, no direct relationship between the kinetics of the increase in TCRgamma(delta)+ i-IELs and T. spiralis expulsion was observed in infected mice. Taken together, these results implicate that gamma(delta) i-IELs probably perform a unique functions related to the regulation of the GC proliferation accompanying T. spiralis gut infection. As is known, these TCRgamma(delta)+ i-IELs may release mediators or growth factors that in turn influence GC differentiation. With the use of dexamethason (DEX), a potent anti-inflammatory agent which also induces apoptotic cell death in i-IELs, we have confirmed that the expulsion of T. spiralis from the mouse gut is accompanied by an inflammatory response. Indeed, the GC are clearly involved in these phenomena, apparently under the regulation by TCRgamma(delta)+ i-IEL-mediated responses, since DEX abrogated GC proliferation in T. spiralis-infected C57BL mice and subsequently augmented adult worm burden. Our data also show that the rejection of adult worms starts concurrently with a significant increase in TCRalpha(beta)+ and CD8+ i-IELs (P < 0.05 and P < or = 0.01, respectively), namely by day 7 p.i. At the same time, CD4+ cells significantly decreased (P < 0.05) in the intestinal epithelium of T. spiralis-infected, vs uninfected mice. These results may indicate that the TCRalpha(beta)+ and CD8+ i-IELs act as effectors of anti-T spiralis defence reactions. The implications of these findings for the potential role of intestinal intraepithelial CD8+ and TCRalpha(beta)+ cells in the pathogenesis of the intestinal lesions during T. spiralis gut infection are discussed. PMID- 10833018 TI - A redescription of the adult male of Caecognathia cryptopais (Barnard, 1925) (Crustacea: Isopoda: Gnathiidae) from southern Africa. AB - A redescription of the adult male of Caecognathia cryptopais (Barnard, 1925) is provided from syntypes and other material deposited in the South African Museum. The generic status of Caecognathia cryptopais is also revised. This redescription is based on light and scanning electron microscopy. PMID- 10833019 TI - Comparison of the protein profiles of salivary gland extracts derived from three species of unfed and partially fed ixodid ticks analysed by SDS-PAGE. AB - Salivary gland extracts (SGE) from unfed and 5 days fed adult female Ixodes ricinus (Linnaeus, 1758); Haemaphysalis inermis (Birula, 1895) and Dermacentor reticulatus (Fabricius, 1794) ticks were prepared. The protein content after feeding increased by 10.6, 8.7 and 6.8 times, respectively. Extracts were equilibrated to the same protein content and submitted to SDS-polyacrylamide gel electrophoresis followed by computer analysis of the scanned gels. Relative differences in protein profiles of extracts obtained from unfed and partially fed ticks were found in all species and some of them were similar in all three species used in the study. Results demonstrate that the increase of the protein content in salivary glands during the feeding does not occur proportionally. Some proteins are synthesised preferentially (67.1 kDa, 13.5 kDa) but other bands (in range of 15-16 kDa) present in the SGE derived from unfed ticks are less discernible in that of fed ticks. PMID- 10833020 TI - Notes on mites of the family Myobiidae (Acari: Prostigmata) parasitising rodents (Mammalia: Rodentia) in Iran. AB - Six mite species of the family Myobiidae, Radfordia (Austromyobia)persica sp. n., Radfordia (Austromyobia) merioni Bochkov, Dubinina et Chirov, 1990, Radfordia (Radfordia) acomys Fain et Lukoschus, 1977, Radfordia (Radfordia) affinis (Poppe, 1896), Radfordia (Graphiurobia) dyromys Fain et Lukoschus, 1973, and Myobia (Myobia) murismusculi (Schrank, 1781) were found in Iran on the rodents Gerbillus cheesmani Thomas, Meriones libycus Lichtenstein, Acomys cahirinus (Desmarest), Alus musculus L., Dryomys nitedula (Pallas), and Mus muscullus, respectively. R. (A.) persica is described as a new species from the female, male and tritonymph. The other five myobiid species are new to Iran. PMID- 10833021 TI - Kabatana gen. n., new name for the microsporidian genus Kabataia Lom, Dykova et Tonguthai, 1999. PMID- 10833022 TI - Detection of spirochetes in, and isolation from, culicine mosquitoes. PMID- 10833023 TI - Rapidly switchable matrix-assisted laser desorption/ionization and electrospray quadrupole-time-of-flight mass spectrometry for protein identification. AB - We describe a new interface for a prototype quadrupole-quadrupole-time-of-flight (TOF) mass spectrometer (Centaur, Sciex) that allows rapid switching between electrospray ionization (ESI) and matrix-assisted laser desorption/ionization (MALDI) modes of operation. Instrument performance in both modes is comparable (i.e., resolution approximately 10,000 FWHM, mass accuracy <10 ppm, sensitivity approximately 1 fmol) because the ion source is decoupled from the TOF mass analyzer by extensive gas collisions in the quadrupole stages of the instrument. The capacity to obtain side-by-side high quality ESI and MALDI mass spectra from a single proteolytic mixture greatly facilitates the identification of proteins and elucidation of their primary structures. Improved strategies for protein identification result from this ability to measure spectra using both ionization modes in the same instrument and to perform MS/MS on singly charged as well as multiply charged ions. Examples are provided to demonstrate the utility and performance of the modified instrument. PMID- 10833024 TI - Characterization of a cross-linked DNA-endonuclease VIII repair complex by electrospray ionization mass spectrometry. AB - Electrospray mass spectrometry techniques were used to characterize components of the active site in Endonuclease VIII by identifying the amino acid sequence and the binding site for a tryptic peptide derived from Endo VIII in a cross-linked DNA-peptide complex. Endo VIII, a DNA repair enzyme with both glycosylase and lyase activities, was covalently bound to a thymidine glycol-containing oligodeoxynucleotide duplex by converting a transient Schiff base formed during the course of the glycosylase activity to a stable covalent bond by chemical reduction with sodium borohydride. After tryptic digestion of the initial product, the identification of the cross-linked peptide was deduced initially from the molecular mass of the tryptic product obtained by negative ion electrospray mass analysis. Nanospray tandem mass spectrometry (MS/MS) analysis of the tryptic product corroborated the molecular mass of the peptide fragment and verified the point of attachment to the oligomer, but failed to produce sufficient fragmentation to sequence the peptide completely. Direct evidence for the amino acid sequence of the peptide was obtained after enzymatic digestion of the DNA portion of the cross-linked DNA-peptide product and analysis by negative ion nanospray MS/MS. Examination of the ions from collision induced fragmentation disclosed that this substance was the N-terminal tryptic fragment of Endo VIII cross-linked to a portion of the oligomer, and that the N-terminal proline from Endo VIII was covalently bound to the residual deoxyribose moiety at the original location of the thymine glycol in the oligomer. PMID- 10833025 TI - Identification of a new metabolite of macrolide immunosuppressant, like rapamycin and SDZ RAD, using high performance liquid chromatography and electrospray tandem mass spectrometry. AB - Previously unknown metabolites from the two macrolide immunosuppressants rapamycin (sirolimus) and SDZ RAD [40-O-(2-hydroxyethyl)rapamycin] obtained after in vitro incubation with human liver microsomes have been purified. Structure elucidation was performed by nanoelectrospray ionization tandem mass spectrometry applying low energy collision activated dissociation. This ionization method is, as shown here, a powerful tool to determine metabolic pathways by analysis of even low abundance products. Product ion spectra of the isolated metabolites indicate a new kind of biotransformation reaction for rapamycin and SDZ RAD. The proposed metabolic pathway starts with an ester hydrolysis which leads to a ring opened structure. A dehydration on C33-C34 and a supplementary hydrogenation at C33-C34 result in a structure similar to the ring-opened isomer with an single bond at C33-C34. PMID- 10833026 TI - Electrospray mass and tandem mass spectrometry identification of ozone oxidation products of amino acids and small peptides. AB - Aqueous ozonation of the 22 most common amino acids and some small peptides were studied by electrospray mass (ESI-MS) and tandem mass spectrometry. After 5 min of ozonation only His, Met, Trp, and Tyr form oxidation products clearly detectable by ESI-MS. For His, the main oxidation product is formed by the addition of three oxygen atoms, His + 30; for Met and Tyr by the addition of one oxygen atom, Met + O and Tyr + O, and for Trp by the addition of two oxygen atoms, Trp + 20. Ozone oxidation occurs rapidly, products are already detected after 30 s of ozonation, and the reactivity order is Met > Trp > Tyr > His. The structures of the oxygen addition products were investigated by electrospray product ion mass spectra, and by comparing these spectra to those of protonated intact amino acids, and when available, to those of model compounds. His + 30 was assigned as 2-amino-4-oxo-4-(3-formylureido)butanoic acid (1) formed by oxidation of the His imidazole ring, Met + O as methionine sulfoxide (2), Trp + 20 as N formylkynurenine (4), and Tyr + O as a mixture of dihydroxyphenylalanines (7 and 8). Ozonation of peptides show that the same number of oxygen atoms are added as expected from the ozonation of the free amino acids. The product ion mass spectra of both the protonated intact peptides, MH+, and the main ozonation products (M + nO)H+ (n = 1-3) revealed b and y type ions as the main fragments, which allow one to assign the type and location of modified amino acid in the model peptides. PMID- 10833027 TI - Endgroup determination of synthetic polymers by electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry AB - Electrospray ionization (ESI) was performed on a Fourier transform ion cyclotron resonance mass spectrometer for the endgroup and monomer mass determination of three poly(oxyalkylene)s in the mass range of 400-8000 Da. A combined use of the multiple charge states observed with ESI, leads to a threefold increase in accuracy of the endgroup and monomer determination. The improvement is attributed to the increased number of datapoints used for the regression procedure, yielding more accurate results. Endgroup masses are determined with a mass error better than 5 and 75 millimass units for the molecular weight range of 400-4200 and 6200 8000 Da, respectively. A mass error of better than 1 millimass unit was observed for all monomer mass determinations. With ESI, endgroup and monomer masses have been determined for poly(ethylene glycol) oligomers with a mass higher than 8000 Da. This is almost two times higher than observed with matrix-assisted laser desorption/ ionization on the same instrument. PMID- 10833028 TI - Gas-phase basicities of the isomeric dihydroxybenzoic acids and gas-phase acidities of their radical cations AB - The thermochemical acid/base properties of the six dihydroxybenzoic acids (x,y DHB) as prototypical matrices used in matrix-assisted laser desorption/ionization (MALDI) have been investigated. The ground-state gas-phase basicities (GB) of the six DHB isomers and the gas-phase acidities (deltaG acid) of the corresponding radical cations ([x,y-DHB]*+) have been determined by Fourier-transform ion cyclotron resonance mass spectrometry employing the thermokinetic method. The gas phase basicities vary from 814 kJ mol-1 for the least basic isomer, 3,5-DHB, to 831 kJ mol-1 for the most basic isomer, 2,4-DHB. The obtained gas-phase acidities of the corresponding radical cations vary from 815 kJ mol-1 for the most acidic species, 3,4-DHB, to 858 kJ mol-1 for the least acidic one, 2,5-DHB. The results indicate that ground-state proton transfer from the matrix radical cations to the analyte may play a role in the ionization process of MALDI, whereas proton transfer from protonated matrix molecules can be excluded. PMID- 10833029 TI - Time-of-flight secondary ion mass spectrometry of matrix-diluted oligo- and polypeptides bombarded with slow and fast projectiles: positive and negative matrix and analyte ion yields, background signals, and sample aging. AB - Human angiotensin II, chain B of bovine insulin, and porcine insulin were determined by time-of-flight secondary ion mass spectrometry under impact of approximately 25 keV Xe+ and SF5+ ion beams and approximately 100 MeV 252Cf fission fragments. Matrix-embedded samples, dissolved in a large surplus of alpha cyano-4-hydroxycinnamic acid, were prepared by nebulizer spray deposition, neat samples by the droplet technique. It is shown that the status of the sample can be assessed by evaluating the matrix-specific features of the mass spectra. The beneficial effect of matrix isolation was small for angiotensin but large for the insulin samples, which did not show parent peaks from neat material. Negative ion yields under SF5+ impact were up to a factor of 50 higher than with Xe+. For positive secondary ions, the enhancement was much smaller. The mass spectra produced by slow ion beams or fast fission fragments were qualitatively similar. Quantitative differences include the following: with fast projectiles the yields were about 10-30 times higher than with slow ions, but similar for negative ion emission under SF5+ bombardment; the analyte-to-matrix yield ratios were higher with slow ions and up to 250 times higher than the molar analyte concentration; for analyte ions the peak-to-background ratios were higher using slow projectiles; the fraction of carbon-rich collisionally formed molecular ions was much higher with fast projectiles. Sample aging in vacuum for up to five weeks strongly reduced the yield of protonated analyte molecules ejected by slow ion impact, but not of deprotonated species. Hence protonation seems to correlate with sample "wetness" or the presence of volatile proton-donating additives. PMID- 10833030 TI - Composition of N-linked carbohydrates from ovalbumin and co-purified glycoproteins. AB - Analysis of commercial samples of chicken ovalbumin by reversed-phase high performance liquid chromatography and matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) showed the presence of several other co-purifying glycoproteins. These were isolated, subjected to tryptic digestion, and two of them were identified as ovomucoid and chicken riboflavin binding-protein following database matching of the peptide masses obtained by MALDI. The N-linked glycans were released from the glycoproteins and their structures were examined by MALDI-MS in combination with exoglycosidase digestion. Ovalbumin was found to be glycosylated mainly with high-mannose and hybrid structures, consistent with profiles obtained on the intact glycoprotein by electrospray. The other glycoproteins contained mainly larger, complex glycans with up to five antennae, many of which had earlier been associated with ovalbumin. PMID- 10833031 TI - Postsource decay fragmentation of N-linked carbohydrates from ovalbumin and related glycoproteins. AB - N-linked glycans were released from chicken ovalbumin by hydrazinolysis and examined by matrix-assisted laser desorption/ionization mass spectrometry. Postsource decay analysis showed that most fragment ions arose as the result of internal glycosidic cleavages involving loss of nonreducing terminal residues from ions that had lost one or both GlcNAc residues from the chitobiose core [GlcNAcbeta(1 --> 4)GlcNAc]. Cross-ring fragments were abundant from the reducing terminal GlcNAc but other cross-ring fragments were weak. The ion found to be most useful for determining the composition of the antennae attached to the 3- or 6-linked core mannose residues was an internal cleavage ion formed by loss of both the chitobiose core and the antenna linked to the 3-position of the core branching mannose. This ion was observed to lose water in the absence of a "bisecting" GlcNAc residue (beta1 --> 4 linked to the core mannose) and to lose a GlcNAc molecule (221 mass units) when a bisecting GlcNAc residue was present. PMID- 10833032 TI - Quantification of 3-nitrotyrosine in biological tissues and fluids: generating valid results by eliminating artifactual formation. AB - Reactive nitrogen species such as peroxynitrite can nitrate specific amino acids, whether free or protein bound, and 3-nitrotyrosine is believed to be one marker of this reaction. To examine the significance of this pathway in biological systems we have developed an accurate, sensitive, and specific assay for 3 nitrotyrosine based on combined liquid chromatography tandem mass spectrometry. Our approach allowed simultaneous analysis of both tyrosine and 3-nitrotyrosine and employs isotopomer standards (i.e., [15N1, 13C9]-tyrosine and [13C6] 3nitrotyrosine). Calibration curves were linear (r2 = 0.999) across the range 0.5 100 pg/microL (i.e., 2.2-442 fmol/microL), and the detection limit for standard samples was 0.5 pg/microL (2.2 fmol/microL, or 10 fmol on column; S/N = 5) or 1 pg/microL (4.4 fmol/microL) for extracted (biological) samples. As a component of this study we have undertaken an extensive investigation of artifactual formation of 3-nitrotyrosine under conditions that exist during sample extraction and derivatization. Our studies show that under appropriate conditions (low pH, elevated temperatures, and in the presence of a vast excess of the two substrates, tyrosine and the nitrate anion), 3-nitrotyrosine can readily be formed as an artifact. PMID- 10833033 TI - Liquid chromatography mass spectrometry with supersonic molecular beams AB - A new approach for liquid chromatography mass spectrometry (LC-MS) is described, based on achieving soft thermal vaporization followed by supersonic expansion and direct sample compound ionization, while in a supersonic molecular beam (SMB). The soft molecular vaporization step utilizes spray formation that is continued by fast thermal vaporization inside a channel supersonic nozzle, followed by ultrafast supercooling in a supersonic expansion. The short time (several microseconds) spent by the vaporized compound in the heated nozzle prior to its expansion cooling may result in incomplete vibrational equilibrium and thus reduced degree of dissociation. In addition, even if vibrational equilibrium at the nozzle temperature is obtained, the sample compounds have significantly reduced time for their dissociation, which is thus further minimized (kinetic consideration). As soon as the molecules expand and form a SMB, they are supercooled and any further dissociation is avoided. While in the SMB, the sample molecules can be ionized either by electron ionization as described in this paper or by hyperthermal surface ionization. The major goal of this method is to obtain high quality library searchable electron ionization mass spectra, for a broad range of thermally labile compounds, with higher sensitivity than that achievable by particle beam LC-MS. The soft thermal vaporization nozzle is described and mass spectral results with corticosterone are demonstrated. The potential advantageous features of this new method are discussed. PMID- 10833035 TI - The American Society for Mass Spectrometry. Year 2000 awards PMID- 10833034 TI - Magnetic field focused ion accumulation for an internal bore liquid chromatography electrospray ionization Fourier transform ion cyclotron resonance mass spectrometer using a central trapping electrode AB - Presented is the application and evaluation of a magnetic field focusing central trapping electrode ion accumulation cell for a capillary liquid chromatography electrospray Fourier transform ion cyclotron (LC-ESI/FTICR) mass spectrometer. The ESI source and accumulation cell are located within the magnetic field to confine the radial motion of the ions, eliminating the need for elaborate focusing optics to transport the ions to the low-pressure analyzer cell for analysis. The central trapping electrode accumulation cell increases sensitivity by providing the necessary potential well in a confined volume to capture ions currently lost during the detection event of LC/FTICR experiments. With this electrode geometry the time needed to gate the ions into the analyzer cell is reduced and pump down delays are minimized. The decreased scan time improves LC resolution and increases the number of mass spectral scans per eluted component while maintaining appropriate base pressures for high performance ESI/FTICR. Results achieved with the central trapping electrode accumulation cell include an effective duty cycle increase from 10% to 40%, a S/N increase by a factor of 30, and a mass resolution increase of 80%. PMID- 10833036 TI - Effects of HIV/AIDS education and training on patient care and provider practices: a cross-cutting evaluation. AB - Initial and continuing training in HIV/AIDS service provision is a critical way to enable the nation's health providers to use state-of-the-art developments and perspectives. Typically, the efficacy of HIV/AIDS training programs is evaluated using assessments administered to trainees immediately following the training. This study reports cross-cutting findings from telephone interviews conducted with 218 trainees an average of 8 months after training. Long-term training effects are examined in three domains: (a) general perspectives on HIV/AIDS; (b) health care provider service provision; and (c) changes in procedures and operations at the health care system level. The findings show the different ways that the training experience had long-term positive and observable effects in these three domains. In some cases, background characteristics and job positions predicted the specific type of reported training effects. The pattern of results suggests ways in which training methods can be targeted to specific audiences. PMID- 10833037 TI - The long-term effectiveness of an HIV prevention intervention for low-income African American women. AB - The effectiveness of an HIV prevention intervention for low-income African American women between the ages of 20 and 44 years was assessed. The hypothesis was that at 3, 6, and 9 months, treatment intervention women would have significantly greater increase from baseline on self-efficacy, condom use knowledge, protective sexual behavior, prevention community behavior, and perceived HIV vulnerability compared with the control intervention women. Using an interrupted time series design, one of two geographically distinct but demographically similar communities was randomly assigned to the treatment intervention. A total of 280 women participated. Hierarchical linear modeling revealed no differences at pretest for the treatment and control women. Compared with control women, treatment women significantly increased self-efficacy, protective sexual behavior, condom knowledge, and prevention community behavior. A slight decline in protective sexual behavior at 9 months may indicate that a linear pattern of change may not be appropriate to describe sexual behavioral change. PMID- 10833038 TI - HIV risk behaviors among Asian drug users in San Francisco. AB - This study identified patterns of drug use and HIV risk behaviors in relation to cultural factors among Asian drug users in San Francisco. A sample of 92 Asian drug users (35 Chinese, 31 Filipino, 26 Vietnamese) who were not currently enrolled in drug treatment programs were recruited through targeted sampling methods and interviewed using a questionnaire with open-ended questions. The study evaluated responses of the participants based on content analyses. Compared with Chinese and Vietnamese, Filipino drug users had engaged in riskier behaviors in terms of injection drug use, having sex while on drugs, and having sex with injection drug users (IDUs). Cultural factors such as stigma of injection drug use and fear of needles were cited as reasons for not injecting drugs. Among IDUs, half cited trust as a reason for having shared needles. AIDS prevention programs targeting Asian drug users should consider specific cultural factors among high-risk groups (i.e., Filipinos, immigrants, women, and IDUs). PMID- 10833039 TI - Psychosocial factors as predictors of unsafe sexual practices among young adults. AB - This study examined the influence of both psychosocial constructs and demographic features on the sexual risk-taking behavior of young adults. Three hundred seventy-four students between the ages of 18 and 29 were drawn from a medium sized university. All participants were assessed using measures that included the AIDS Coping and Change Survey, the AIDS Psychosocial Scale, and the AIDS Knowledge and Attitudes Survey. Descriptive statistics were calculated for levels of HIV risk behavior (failure to use condoms combined with casual or anonymous sexual partners), HIV/AIDS knowledge, alcohol and drug use, sexual history, and other demographic features. Factor analysis was then used to examine the latent factors associated with HIV risk behavior. All confirmed factors and demographic items that were shown to be significantly correlated with HIV risk behavior were then examined for predictive ability using a regression analysis. Ten factors or variables were found to be predictive of HIV risk behavior, including a Developmental Factor which was found to be moderately predictive of HIV risk behavior. Other predictors included Resistance to Change in response to AIDS; Perceived Risk of exposure, the influence of peer groups, age, alcohol use, marijuana use; a history of infidelity; a younger age of sexual initiation; and a lower self-efficacy. PMID- 10833040 TI - Fear and violence: raising the HIV stakes. AB - Through focus group methodology, the study examines three contexts that delineate the co-occurrence of intimate partner violence and sexual risk behaviors among 68 women on methadone. First, it explores the ways in which the presence of physical abuse in an intimate relationship prevents women from asking their partners to use a condom. Second, it describes the ways in which the couple's drug involvement increases the risk of physical and sexual violence, and concomitant sexual HIV risks. Third, it discusses the context in which sexual assault and rape occur in these established intimate relationships and how these abusive events increase women's risks of becoming infected with HIV. The research is guided by feminist theory, which affords powerful insight into the contexts in which women are put at risk for HIV and partner violence. The study provides a discussion on the implications of the findings to HIV prevention for women who are risk for both HIV and partner violence. PMID- 10833041 TI - Acceptability of a stage-matched expert system intervention to increase condom use among women at high risk of HIV infection in New York City. AB - There is an urgent need to develop and implement effective methods for sexual behavior change to curb the spread of HIV. Condoms remain one of the most effective strategies for achieving this, yet consistent condom use is generally low, especially among those at highest risk. This article describes the acceptability of an interactive computer-based expert system designed to increase condom use in women at high risk of HIV infection. The expert system is based on the transtheoretical stages of change model. Using a computer, participants respond to questions about their attitudes and behavior toward using condoms and receive immediate feedback which is matched to their readiness to use condoms. The women were found to be at all stages of change for condom use, although a large proportion of the women (42%) were at early stages of behavior change because they were considering but not using condoms every time during sex with men. The expert system was found to be acceptable to this high-risk group of women. They almost unanimously agreed that they found the feedback useful, would return to use the system again, and would recommend it to a friend. These findings indicate that traditional intervention strategies which assume individuals are ready to use condoms consistently would be appropriate for only about one third of these women, underscoring the importance and potential utility of stage-matched interventions. PMID- 10833042 TI - The Wilhelmine E. Key 1999 Invitational lecture. Predicting the evolution of human influenza A. AB - We studied the evolution of the HA1 domain of the H3 hemagglutinin gene from human influenza virus type A. The phylogeny of these genes showed a single dominant lineage persisting over time. We tested the hypothesis that the progenitors of this single evolutionarily successful lineage were viruses carrying mutations at codons at which prior mutations had helped the virus to avoid human immune surveillance. We found evidence that eighteen hemagglutinin codons appeared to have been under positive selection to change the amino acid they encoded in the past. Retrospective tests show that viral lineages undergoing the greatest number of mutations in the positively selected codons were the progenitors of future H3 lineages in nine of eleven recent influenza seasons. Codons under positive selection were associated with antibody combining sites A or B or the sialic acid receptor binding site. However, not all codons in these sites had predictive value. Monitoring new H3 isolates for additional changes in positively selected codons might help identify the most fit extant viral strains that arise during antigenic drift. PMID- 10833043 TI - Genomic ancestry of the American puma (Puma concolor). AB - Puma concolor, a large American cat species, occupies the most extensive range of any New World terrestrial mammal, spanning 110 degrees of latitude from the Canadian Yukon to the Straits of Magellan. Until the recent Holocene, pumas coexisted with a diverse array of carnivores including the American lion (Panthera atrox), the North American cheetah (Miracynonyx trumani), and the saber toothed tiger (Smilodon fatalis). Genomic DNA specimens from 315 pumas of specified geographic origin (261 contemporary and 54 museum specimens) were collected for molecular genetic and phylogenetic analyses of three mitochondrial gene sequences (16S rRNA, ATPase-8, and NADH-5) plus composite microsatellite genotypes (10 feline loci). Six phylogeographic groupings or subspecies were resolved, and the entire North American population (186 individuals from 15 previously named subspecies) was genetically homogeneous in overall variation relative to central and South American populations. The marked uniformity of mtDNA and a reduction in microsatellite allele size expansion indicates that North American pumas derive from a recent (late Pleistocene circa 10,000 years ago) replacement and recolonization by a small number of founders who themselves originated from a centrum of puma genetic diversity in eastern South America 200,000-300,000 years ago. The recolonization of North American pumas was coincident with a massive late Pleistocene extinction event that eliminated 80% of large vertebrates in North America and may have extirpated pumas from that continent as well. PMID- 10833044 TI - Origins and divergence times of mammalian class II MHC gene clusters. AB - The class I and II major histocompatibility complex (MHC) genes are apparently subject to evolution by a birth-and-death process. The rate of gene turnover is much slower in the latter genes than in the former. In placental mammals, the class II region can be subdivided into different orthologous subregions or gene clusters (DR, DQ, DO, and DN), but the origins and evolutionary relationships of these gene clusters are not well established. Here we report the results of our study of the times of origin and evolutionary relationships of these gene clusters in mammals. Our analysis suggests that both class II alpha-chain and beta-chain gene clusters are shared by placental mammals and marsupials, but the gene clusters from nonmammalian species are paralogous to mammalian gene clusters. We estimated the times of divergence between gene clusters in placental mammals using the linearized tree and distance regression methods. Our results indicate that most gene clusters originated 170-200 million years (MY) ago, but that DO beta-chain genes diverged from the other beta-chain gene clusters approximately 210-260 MY ago. The phylogenetic trees for the alpha- and beta chain genes were not congruent, suggesting that the evolutionary history of the class II gene clusters is more complex than previously thought. PMID- 10833045 TI - Evolution of RH genes in hominoids: characterization of a gorilla RHCE-like gene. AB - The human RH locus is responsible for the expression of the Rh blood group antigens. It consists of two closely linked genes, RHD and RHCE, that exhibit 92% similarity between coding regions. These observations suggest that they are derived from a relatively recent duplication event. Previously a study of nonhuman primate RH-like genes demonstrated that ancestral RH gene duplication occurred in the common ancestor of man, chimpanzees and gorillas. By amplification of intron 3 and intron 4 of gorilla RH-like genes, we have now shown that, like man, gorillas possess two types of RH intron 3 (RHCE intron 3 being 289 bp longer than the RHD intron 3) and two types of intron 4 (RHCE intron 4 being 654 bp longer than the RHD intron 4). Here we report the characterization of a cDNA encoded by a gorilla RH-like gene which possesses introns 3 and 4 of the RHCE type. A comparison of this gorilla RHCE-like coding sequence with previously characterized human and ape cDNA sequences suggests that RH genes experienced complex recombination events after duplication in the common ancestor of humans, chimpanzees and gorillas. PMID- 10833046 TI - Gene diversity of chimpanzee ABO blood group genes elucidated from intron 6 sequences. AB - The human and nonhuman primate ABO blood group gene shows relatively large numbers of nucleotide differences around the exon 7 region. In this study we determined intron 6 sequences for 9 alleles of common chimpanzee and for 3 alleles of bonobo to estimate nucleotide diversities among them. Sequence length polymorphisms are observed in this region as a repeat appears one to five times. From a phylogenetic network of intron 6 sequences of ABO blood group genes for humans, common chimpanzee, and bonobo, parallel substitutions and/or some kinds of convergent events are predicted in the chimpanzee lineage. We also estimated nucleotide diversities for common chimpanzee and bonobo ABO blood group genes; these values were 0.219% and 0.208%, respectively. PMID- 10833047 TI - Color vision of the coelacanth (Latimeria chalumnae) and adaptive evolution of rhodopsin (RH1) and rhodopsin-like (RH2) pigments. AB - The coelacanth, a "living fossil," lives at a depth of about 200 m near the coast of the Comoros archipelago in the Indian Ocean and receives only a narrow range of light at about 480 nm. To see the entire range of "color" the Comoran coelacanth appears to use only rod-specific RH1 and cone-specific RH2 visual pigments, with the optimum light sensitivities (lambda max) at 478 nm and 485 nm, respectively. These blue-shifted lambda max values of RH1 and RH2 pigments are fully explained by independent double amino acid replacements E122Q/A292S and E122Q/M207L, respectively. More generally, currently available mutagenesis experiments identify only 10 amino acid changes that shift the lambda max values of visual pigments more than 5 nm. Among these, D83N, E1220, M207L, and A292S are associated strongly with the adaptive blue shifts in the lambda max values of RH1 and RH2 pigments in vertebrates. PMID- 10833048 TI - Pseudogene evolution and natural selection for a compact genome. AB - Pseudogenes are nonfunctional copies of protein-coding genes that are presumed to evolve without selective constraints on their coding function. They are of considerable utility in evolutionary genetics because, in the absence of selection, different types of mutations in pseudogenes should have equal probabilities of fixation. This theoretical inference justifies the estimation of patterns of spontaneous mutation from the analysis of patterns of substitutions in pseudogenes. Although it is possible to test whether pseudogene sequences evolve without constraints for their protein-coding function, it is much more difficult to ascertain whether pseudogenes may affect fitness in ways unrelated to their nucleotide sequence. Consider the possibility that a pseudogene affects fitness merely by increasing genome size. If a larger genome is deleterious--for example, because of increased energetic costs associated with genome replication and maintenance--then deletions, which decrease the length of a pseudogene, should be selectively advantageous relative to insertions or nucleotide substitutions. In this article we examine the implications of selection for genome size relative to small (1-400 bp) deletions, in light of empirical evidence pertaining to the size distribution of deletions observed in Drosophila and mammalian pseudogenes. There is a large difference in the deletion spectra between these organisms. We argue that this difference cannot easily be attributed to selection for overall genome size, since the magnitude of selection is unlikely to be strong enough to significantly affect the probability of fixation of small deletions in Drosophila. PMID- 10833049 TI - Inheritance of UV-B tolerance in seven ecotypes of Arabidopsis thaliana L. Heynh. and their F1 hybrids. AB - We used a partial diallel mating design to examine morphologic response to supplementary ultraviolet-B (UV-B) radiation of seven ecotypes of Arabidopsis thaliana L. Heynh. from several geographic locations in Europe. We were particularly interested in the inheritance of UV-B tolerance by the F1 generation. Morphologic traits included plant height, rosette diameter, number of shoots (lateral branches from the rosette) and branches (lateral branches above the rosette), and reproductive and vegetative dry mass. To effect a large difference in UV treatments, plants under treatment received 11 kJ/m2/day of biologically effective UV-B radiation while control plants received no UV-B radiation. Genotype effects were observed for all traits (P < .0001), but a significant treatment effect and genotype x treatment interactions were detected only for plant height (P = .0001), rosette diameter (P = .0229), and vegetative (P = .0260) and reproductive dry mass (P = .0900). General combining ability was significant for plant height (P < .0001) and vegetative mass (P = .0563), whereas specific combining ability was significant for rosette diameter (P = .0220) and vegetative mass (P = .0506). These results suggest that both pure lines and hybrids of Arabidopsis can be developed for greater tolerance of UV-B radiation. Similar findings for crop species might lead to the development of UV tolerant varieties. PMID- 10833050 TI - Penetrance and expressivity of the gene for double podding in chickpea. AB - The double-pod per peduncle trait is known to contribute to increased seed yield in chickpea (Cicer arietinum L.). A cross was made between the single-podded variety ICCV 2 and the double-podded variety JG 62 in 1993. Penetrance and expressivity of the gene for double podding was studied in an F2 population and F10 recombinant inbred lines (RILs) of this cross. Homozygous recessive allele of this gene (ss) governs the production of double flowers and pods per peduncle. Results indicated that the s allele has unstable penetrance and variable expressivity. The penetrance of this allele was 53% for the F2 and 84.5% for the RILs. The ranges for the expression of this trait among the penetrant F2 individuals and the penetrant RILs were 1.1-14.8% and 0.1-33.0%. These were 8.3 30.8% for early sown and 17.1-68.7% for the late sown double-podded parent JG 62. Thus it appears that the allele shows greater penetrance and enhanced expressivity under soil moisture stress. In the F2 the seed yield advantage of the double-podded over the single-podded plants was 18%, whereas among the RILs it was 7%. The increased number of pods and seeds contributed to the higher yield. However, there was a slight decrease in seed size of the double-podded genotypes. An increase in the size of seed may have a role in the decreased penetrance and expressivity of this allele among the double-podded segregants of the ICCV 2 x JG 62 chickpea cross. PMID- 10833051 TI - White yellow virescent pattern in winter rye: inheritance, plant growth, and ultrastructure of plastids. AB - Inbred lines from different varieties of cultivated plants characterized by a white yellow irregular pattern on the leaves obtained after selection in the inbred generation (S3) of winter rye (Secale cereale L.) were the object of the present studies. The feature of a white yellow irregular pattern in all lines was monomeric and recessive. This trait in L158b, wch, and zp was determined by the same recessive gene marked with the symbol wyv1, "white yellow virescent." The gene responsible for the appearance of the above feature in line L24 was nonallelic to the gene wyv1, therefore it was designated as the sequent gene of the same series--wyv2. The studied forms of plants were characterized by a diminution in the number of plastids and in chlorophyll (a plus b) content in mesophyll cells of leaves. Contrary to typical ultrastructure of chloroplasts in dark green plants (control), plastids in lines with the white yellow virescent pattern on the leaves showed variations in ultrastructure from numerous granal and intergranal thylakoids to a reduced number. PMID- 10833052 TI - Novel flowering time variation in the resynthesized polyploid Brassica napus. AB - Recent molecular data using resynthesized polyploids of Brassica napus established that genome changes can occur rapidly after polyploid formation. In this study we present data that de novo phenotypic variation for flowering time also occurs rapidly after polyploidization. Two initial polyploid plants were developed by reciprocal crosses of B. rapa and B. oleracea followed by chromosome doubling to establish two lineages, each of which was expected to be homozygous and homogeneous. Several sublineages of each lineage were advanced by self pollination. The range in days to flower of the sixth generation plants was 39-75 and 43-64 for the two lineages. Analysis of seventh generation progeny indicated that the variation was heritable. Lines were selected and self-pollinated to the eighth generation and also testcrossed to a natural B. napus cultivar; the testcross plants were then self-pollinated. Differences in flowering time were also inherited in these advanced generations. Days to flower was significantly correlated with leaf number in each generation. The rapid evolution of new phenotypic variation, like that observed in this model system, may have contributed to the success and diversification of natural polyploid organisms. PMID- 10833053 TI - Molecular phylogeny of Philippine freshwater sardines based on mitochondrial DNA analysis. AB - The commercially important Sardinella species (family Clupeidae or herrings) usually thrive in marine environments. An exception is Sardinella tawilis of Taal Lake, Batangas, Philippines, the only known freshwater sardine. This species is believed to have immigrated from Balayan Bay to the lake when it was formed in the course of volcanic eruptions some 240 years ago. To determine the relationship of S. tawilis to the marine species S. albella, S. fimbriata, and S. longiceps from the Balayan Bay we sequenced 358 bp of the cytochrome b gene and the mitochondrial control region. The cytochrome b gene was highly conserved and contained little phylogenetic information. The control region sequences, however, demonstrated two highly diversified main haplotypes grouping S. tawilis with S. albella, as shown by maximum parsimony and neighbor-joining analysis. The haplotypes are characterized by the presence of an 81 bp indel and up to eight 35 bp tandem repeat elements. The repeat copy number varied within individuals of S. tawilis and S. albella, thus showing heteroplasmy in these two species only. The analysis of two subpopulations of S. tawilis revealed restricted substitutions that may indicate the beginning of genetic differentiation of the two subpopulations. PMID- 10833054 TI - Salmon: a new autosomal mutation demonstrating incomplete dominance in the boine snake Boa constrictor. AB - An unusual and attractive pigmentation pattern mutation termed "salmon" has been identified in the United States in several captive colonies of the common neotropical boine snake boa constrictor [Boa constrictor (Boidae)]. Boa constrictors expressing the Sa pigmentation pattern appear to be restricted to regions of Panama. Animals with the Sa phenotype exhibit a sharp decrease in melanophore pigments (e.g., melanin) and an increase of xanthophore pigments (e.g., pteridines and carotenoids) throughout the body, including ventral and caudal regions. Moreover, the dorsal saddles (blotches) and lateral diamond patterns are greatly reduced and/or absent. Our study was initiated using a female B. constrictor born in captivity and expressed the Sa pigmentation pattern. Results from breeding experiments indicated an inheritance mode of autosomal incomplete dominance for the Sa and Wt alleles. PMID- 10833055 TI - Physical mapping of rRNA genes in Medicago sativa and M. glomerata by fluorescent in situ hybridization. AB - Fluorescent in situ hybridization (FISH) was applied to diploid and tetraploid subspecies of alfalfa (Medicago sativa L.) to investigate the distribution of rRNA genes and to utilize the sites of 18S-5.8S-25S rDNA and 5S rDNA sequences as markers for studying the genome evolution within the species. Medicago glomerata Balb., the species considered to be the ancestor of alfalfa, was included in this study in order to obtain more information on the phylogenetics of alfalfa. Simultaneous in situ hybridization was performed with the probes pTa71 and pXVI labeled with digoxigenin and biotin, respectively. In the diploid taxa, M. glomerata, M. sativa ssp. coerulea Schmalh and ssp. falcata Arcangeli, the 18S 5.8S-25S rDNA sequences were mapped to two sites corresponding to the secondary constrictions of the nucleolar chromosome pair, while 5S rDNA appeared to be distributed in two pairs of sites. Chromosomes carrying 5S loci could be distinguished on the basis of their morphological characteristics. The number of rDNA sites detected in the tetraploid M. sativa ssp. falcata and ssp. sativa (L.) L. & L. were twice the number found in the respective diploid ssp. falcata and ssp. coerulea. The results of this study show that the distribution of ribosomal genes was maintained during the evolutionary steps from the primitive diploid to the cultivated alfalfa. Modifications of the number of rRNA loci were not observed. The importance of in situ hybridization for improving karyotype analysis in M. sativa L. is discussed. PMID- 10833056 TI - Karyotype analysis, banding, and fluorescent in situ hybridization in the scarab beetle Gymnopleurus sturmi McLeay (Coleoptera Scarabaeoidea : Scarabaeidae). AB - Conventional staining, differential banding, and in situ hybridization with both ribosomal and telomeric probes to mitotic chromosomes of Gymnopleurus sturmi (Scarabaeoidea : Scarabaeidae) are described. The karyotype is distinguished by a pericentric inversion polymorphism in chromosome 3, which is either acrocentric or subtelocentric. Silver staining (Ag-NOR) and chromomycin A3 (CMA3), failed to study the detection of nucleolar organizer regions (NORs), due to the extensive silver and CMA3 stainability of all GC-rich heterochromatin. Fluorescent in situ hybridization (FISH) using a Paracentrotus lividus (Echinodermata) rDNA probe mapped the ribosomal RNA genes (rDNA). FISH with the all-human telomeric sequences (TTAGGG)n revealed a lack of homology between the telomeric probe and the telomeres of G. sturmi. This suggests that the telomeric hesanucleotide (TTAGGG)n is not so conserved within eukaryotes as it has been hypothesized. PMID- 10833057 TI - Gender differences in acute CNS trauma and stroke: neuroprotective effects of estrogen and progesterone. AB - Increasing evidence has demonstrated striking sex differences in the pathophysiology of and outcome after acute neurological injury. Lesser susceptibility to postischemic and posttraumatic brain injury in females has been observed in experimental models. Additional evidence suggests this sex difference extends to humans as well. The greater neuroprotection afforded to females is likely due to the effects of circulating estrogens and progestins. In fact, exogenous administration of both hormones has been shown to improve outcome after cerebral ischemia and traumatic brain injury in experimental models. The neuroprotection provided by periinjury administration of these hormones extends to males as well. The mechanisms by which estrogen and progesterone provide such neuroprotection are likely multifactorial, and probably depend on the type and severity of injury as well as the type and concentration of hormone present. Both genomic and nongenomic mechanisms may be involved. Estrogen's putative effects include preservation of autoregulatory function, an antioxidant effect, reduction of A beta production and neurotoxicity, reduced excitotoxicity, increased expression of the antiapoptotic factor bcl-2, and activation of mitogen activated protein kinase pathways. It is hypothesized that several of these neuroprotective mechanisms can be linked back to estrogen's ability to act as a potent chemical (i.e., electron-donating) antioxidant. Progesterone, on the other hand, has a membrane stabilizing effect that also serves to reduce the damage caused by lipid peroxidation. In addition, it may also provide neuroprotection by suppressing neuronal hyperexcitability. The following review will discuss experimental and clinical evidence for sex differences in outcome after acute brain trauma and stroke, review the evidence implicating estrogens and progestins as mediators of this neuroprotection following acute neurological injury, and finally, address the specific mechanisms by which these hormones may protect the brain following acute neurological injury. PMID- 10833058 TI - Dissociation of cerebral glucose metabolism and level of consciousness during the period of metabolic depression following human traumatic brain injury. AB - Utilizing [18F]fluorodeoxyglucose positron emission tomography (FDG-PET), we studied the correlation between CMRglc and the level of consciousness within the first month following human traumatic brain injury. Forty-three FDG-PET scans obtained on 42 mild to severely head-injured patients were quantitatively analyzed for the determination of regional cerebral metabolic rate of glucose (CMRglc). Reduction of cerebral glucose utilization, defined as a CMRglc of < or =4.9 mg/100 g/min, was present regionally in 88% of the studies. The prevalence of global cortical CMRglc reduction was higher in severely head-injured patients (86% versus 67% mild-moderate), although the absolute magnitude was similar across the injury severity spectrum (mean CMRglc 3.9 +/- 0.6 mg/100 g/min). The level of consciousness, as measured by the Glasgow Coma Scale, correlated poorly with the global cortical CMRglc value (r = 0.08; p = 0.63). With regards to severity of head injury, this correlation was worst for the severely injured (r = -0.11; p = 0.58) and better for the mildly injured patients (r = 0.50; p = 0.07). In most cases, intraparenchymal hemorrhagic lesions were associated with either focal CMRglc reduction or elevation. It is concluded that the etiologies of CMRglc reduction are likely multifactorial given the complex nature of traumatic brain injury and that the reduction of CMRglc represents a fundamental pathobiologic state following head injury that is not tightly coupled to level of consciousness. PMID- 10833059 TI - Early predictors of mortality and morbidity after severe closed head injury. AB - Mortality and morbidity of 158 patients with severe head injury were studied in relation to age, and early (24-h) clinical and computed tomography data. For comparison of outcome data in survivors, a group of 32 patients with traumatic injuries to parts of the body other than the head was used as controls. Within the head-injured group, the mortality rate was 51%. Logistic regression analyses combined 13 out of 16 predictors into a model with an accuracy of 93%, a sensitivity of 90%, and a specificity of 95%. These include age, Glasgow Coma Scale (GCS) score, pupillary reactivity, blood pressure, intracranial pressure, blood glucose, platelet count, body temperature, cerebral lactate, and subdural, intracranial, subarachnoid, and ventricular hemorrhage. At 6 months postinjury, head-injury survivors and trauma controls were evaluated with the Glasgow Outcome Scale (GOS), a neuropsychological test battery and the Sickness Impact Profile (SIP). Head-injury survivors had a higher proportion of disabilities and neuropsychological dysfunctions than trauma controls. They also report more quality of life-related functional limitations on the SIP scales for mobility, intellectual behavior, communication, home management, eating, and work. Linear regression analysis resulted in age being the only important predictor of outcome on the GOS, the GCS score being the best predictor of neuropsychological functioning, and pupillary reactivity being the most predictive for self-reported quality of life as measured by SIP. Those factors important for predicting mortality (clinical variables such as ICP or blood glucose level, and CT observations) failed to show any significant relationship with morbidity. PMID- 10833060 TI - Astrocytes generate isoprostanes in response to trauma or oxygen radicals. AB - Previous studies have shown that oxygen radical scavengers prevent the reduced cerebral blood flow that occurs following experimental traumatic brain injury. The exact chemical species responsible for the posttraumatic reduction in flow is unknown. We tested whether isoprostanes, which are formed by non-cyclooxygenase dependent free radical attack of arachidonic acid and are vasoconstrictors of the cerebral circulation, are increased in astrocytes following stretch-induced trauma or injury with a free radical generating system. Isoprostane (8-epi prostaglandin F2alpha) was analyzed in cells and in media by immunoassay. Confluent rat cortical astrocytes in culture were injured by a hydroxyl radical generating system consisting of hydrogen peroxide and ferrous sulfate or by rapid stretch of astrocytes grown on a deformable silastic membrane. Some cells were treated with the iron chelator deferoxamine for 1 h before injury. The hydroxyl generating system caused free and cell-bound isoprostanes to increase to more than 400% of control. After trauma, free and membrane bound isoprostanes increased to 321 +/- 34% and 229 +/- 23% of control, respectively, and posttraumatic increases were prevented by deferoxamine. Since astrocytes are in close proximity to cerebral vessels, posttraumatic free radical formation may increase the formation of isoprostanes, which in turn produce vasoconstriction and decrease cerebral blood flow. PMID- 10833061 TI - Three months of chronic ethanol administration and the behavioral outcome of rats after lateral fluid percussion brain injury. AB - This study examined the effects of 3 months of chronic ethanol administration (CEAn) on the behavioral outcome in rats after lateral fluid percussion (FP) brain injury. Rats were given either an ethanol liquid diet (ethanol diet groups) or a pair-fed isocaloric sucrose control diet (control diet groups) for 3 months. Then, rats from both diet groups were subjected to either lateral FP brain injury of moderate severity (1.8 atm) or to sham operation. Postinjury behavioral measurements revealed that brain injury caused significant spatial learning disability in both diet groups. There were no significant differences in spatial learning ability in the sham or brain-injured animals between the control and ethanol diets. However, a trend towards cognitive impairment in the sham animals and a trend towards reduced deficits in the brain-injured animals were observed in the ethanol diet group. Histologic analysis of injured animals from both diet groups revealed similar extents of ipsilateral cortical and hippocampal CA3 damage. These results, in general, suggest that 3 months of CEAn does not significantly alter the behavioral and morphologic outcome of experimental brain injury. PMID- 10833062 TI - Cyclosporin A affects axons and macrophages during Wallerian degeneration. AB - A traumatic injury of a peripheral nerve leads to Wallerian degeneration. It includes the recruitment of macrophages and the phagocytosis of myelin and the remnants of axons. We have previously studied the recruitment of macrophages and now wished to determine if the immunosuppressant cyclosporin A (CsA) affects the number of macrophages at the site of nerve injury. The primary target of CsA is T cells, but it may also have an effect on mononuclear phagocytes which exert a key role during Wallerian degeneration. Rats were divided into two groups: CsA treated animals and control animals. Following transection of the sciatic nerve in the treatment group, the animals received 5 mg/kg CsA subcutaneously. The groups were further subdivided into a freely regenerating nerve group and a sutured nerve group. The number of macrophages and MHC class II positive cells were counted 3 days, 7 days, 2 weeks, 4 weeks, and 8 weeks posttransection; also CD4, CD8, IL-2 receptor positive cells, B cells, and the axonal sprouting were studied. In the CsA-treated group, there were more macrophages in the distal areas under 8 weeks than in the controls (p < 0.05); thus, the clearance of macrophages is delayed in the CsA-treated rats compared to the control rats. In the proximal area, the difference in macrophage number did not gain statistical significance. Additionally, CsA retarded axonal degeneration. CsA affects number of macrophages during Wallerian degeneration, while retarding axonal degeneration and subsequent reinnervation. Its mechanism of action appears to involve either direct or indirect via T-cells-mediated responses. PMID- 10833063 TI - Microdialysis-based long-term measurements of energy-related metabolites in the rat brain following a fluid percussion trauma. AB - The aim of the study was to evaluate an experimental approach based on a fluid percussion rat trauma model in combination with the microdialysis technique for the analysis of cerebral interstitial biochemical alterations following head trauma, and to test the hypothesis that the previously observed acute accumulation of lactate and increase in the lactate pyruvate ratio may persist for several days following trauma. We analyzed how lactate, pyruvate, and glucose were altered in the cortex adjacent to the contusion and in the contralateral side of the brain following a traumatic brain injury. The results were compared with those from sham-operated animals. The lactate concentration in the cortex adjacent to the contusion was 0.73 +/- 0.13 mmol/L and 0.71 +/- 0.08 mmol/L 24 and 48 h posttrauma, respectively, and 0.42 +/- 0.07 mmol/L in the sham group (p < 0.05). The lactate/pyruvate ratio of 18.3 +/- 2.3 in the cortex adjacent to the contusion 24 h posttrauma was higher than corresponding value of 10.3 +/- 1.5 in the sham group (p < 0.05). The lactate/pyruvate ratio 48 h posttrauma did not differ from that in the sham group. Interstitial glucose in the cortex adjacent to the contusion and the sham group were similar. Microdialysis measurements from the contralateral side did not differ from those in the sham group. We conclude that the previously observed acute alterations in brain metabolism persist for at least 48 h posttrauma. Further, the measured parameters from the contralateral side can be used as controls since they did not differ from the sham group. Combining microdialysis with a fluid percussion trauma model may be a tool to explore secondary brain injury mechanisms and evaluate new therapies for the treatment of traumatic brain injury. PMID- 10833064 TI - HLA class II alleles associated with susceptibility and resistance to Crohn's disease in the Jewish population. AB - Previous studies have suggested that susceptibility to Crohn's disease (CD) is associated with the histocompatibility complex (HLA) class II alleles DR1, DQ5, and DR13 in the Caucasian population, DR7 in the French and German populations, and DR4 and DQ4 in the Japanese population. However, little is known about the relationship between HLA class II alleles and CD in the Jewish population since these previous studies included few Jewish individuals. In order to determine whether the HLA associations observed with predominantly non-Jewish populations were also present in the Jewish CD population and whether there were any HLA class II alleles uniquely associated with CD in the Jewish population, 132 CD patients, of which 82 were Ashkenazi Jewish, were HLA-typed using serologic and DNA methods. Ethnically matched controls were similarly typed. No association with DR1 or DR13 was observed in the Jewish CD population although an association with DR13 (OR [odds ratio] = 5.3, p = 0.02) was observed in the non-Jewish CD population. However, an association with DR15 (OR = 2.7, p = 0.03), which is normally associated with ulcerative colitis, was observed in the Jewish, but not non-Jewish, CD group. In addition, a strong negative association was observed with DR3, which was especially striking in the Jewish population (OR = 0.35, p = 0.025); similar negative associations with DR3 have been observed by others using non-Jewish populations. Furthermore, a significant negative association with DR7 (OR = 0.45, p = 0.04) was observed in the Jewish, but not non-Jewish, population. Consistent with this was the negative association with DQ2 (OR = 0.38, p = 0.005), which is in strong linkage disequilibrium with both DR3 and DR7, in the Jewish, but not non-Jewish, population. These studies support previous suggestions that susceptibility to CD in Jewish and non-Jewish populations is determined by distinct genes and provide further support to the hypothesis that a gene on the DR3 haplotype may protect against CD. Furthermore, protection is conferred by the same or another gene found on Jewish, but not non-Jewish, DR7 haplotypes. PMID- 10833065 TI - Nutritional supplementation with N-3 fatty acids and antioxidants in patients with Crohn's disease in remission: effects on antioxidant status and fatty acid profile. AB - In patients with Crohn's disease (CD), malnutrition is frequently observed and is generally accepted to be an important issue. The aim of this study was to investigate the effects of 3 months of supplementation with a liquid formula containing either antioxidants (AO) or n-3 fatty acids plus AO on the antioxidant status and fatty acid profile of plasma phospholipids and adipose tissue, respectively, in patients with long-standing CD currently in remission. In a randomized, double-blind placebo-controlled study, CD patients received either placebo, AO, or n-3 fatty acids plus AO for 3 months in addition to their regular diet. In all, 25/37 CD patients completed the study. AO status was assessed by blood biochemical parameters. A statistical per-protocol analysis was performed. Serum concentrations of selenium, vitamin C, and vitamin E, the activity of superoxide dismutase and total antioxidant status were significantly (p < 0.05) increased after AO supplementation. Furthermore, compared with controls, serum concentrations of beta-carotene, selenium, and vitamin C and the activity of glutathione peroxidase (GPx) were significantly (p < 0.05) lower before supplementation; however, after AO supplementation these levels were not significantly different from controls (except for GPx). N-3 fatty acids plus AO supplementation significantly (p < 0.05) decreased the proportion of arachidonic acid, and increased the proportion of eicosapentanoic acid and docosahexanoic acid in both plasma phospholipids and adipose tissue. Supplementation with antioxidants improved antioxidant status in patients with CD in remission. In addition, supplementation with n-3 fatty acids plus antioxidants significantly changed the eicosanoid precursor profile, which may lead to the production of eicosanoids with attenuated proinflammatory activity. This study indicates that an immunomodulating formula containing n-3 fatty acids and/or AO may have the potential to play a role in the treatment of CD. PMID- 10833066 TI - Short- and long-term follow-up of treatment for lymphocytic and collagenous colitis. AB - Collagenous colitis and lymphocytic colitis cause chronic watery diarrhea. Multiple therapies have been found to improve symptoms but there have been few long-term follow-up studies. Our goal was to obtain long-term clinical follow-up on a cohort of patients with independently confirmed typical histopathologic changes. Pathology slides from 32 cases of collagenous or lymphocytic colitis patients from 1988-1992 were independently reviewed. Twenty-five cases were confirmed by both groups of pathologist as collagenous or lymphocytic colitis. For these 25 patients, charts were reviewed and telephone follow-up interviews were performed in 1992 and 1995. Seven of 32 (22%) of the original cases were not confirmed on independent pathologic interpretation. A 15.8% discordance rate was found between the different groups of pathologists. Patient demographics were similar to previously published reports except one-half of our patients had diarrhea of only 6 months or less. Eighty-one percent of patients receiving 5-ASA agents reported improvement as well as 100% of those receiving prednisone. At 23 month follow-up 86% of patients reported improvement in diarrhea and only 32% required routine medications. At 47 month follow-up all patients reported improved diarrhea and only 29% required routine medications. Collagenous and lymphocytic colitis can sometimes be identified in patients with relatively brief duration diarrhea. Clinical parameters and response to therapy are similar for collagenous or lymphocytic colitis. Most patients with lymphocytic and collagenous colitis improve with therapy such as 5-ASA preparations or steroids. Over a follow-up period of several years, most patients have improvement in diarrhea and generally do not require maintenance medications. Independent pathologic confirmation of the diagnosis should be obtained in patients not responding to therapy. PMID- 10833067 TI - Analysis of a positional candidate gene for inflammatory bowel disease: NRAMP2. AB - Genome scans have identified a region spanning 40 cM on the long arm of chromosome 12 as a susceptibility locus for inflammatory bowel disease (IBD). This locus contains several candidate genes for IBD, one of which is the gene for the natural resistance associated macrophage protein 2 (NRAMP2). This protein is a divalent cation transporter and is expressed in many cells and tissues. The putative role of this protein in innate immunity prompted us to investigate a possible relationship between NRAMP2 and IBD. We assessed the frequency of four restriction fragment length polymorphisms (IVS2+11A/G, IVS4+44C/A, 1254T/C, and IVS15Ex16-16C/G) in a group of 155 Crohn's disease (CD) patients, 114 ulcerative colitis (UC) patients, and 189 healthy controls. Linkage analysis was performed in a group of 70 families with multiple members suffering from IBD. We searched for additional intragenic markers and mutations by sequence analysis of the natural resistance-associated macrophage 2 gene of 33 CD patients, with a positive family history for IBD. We identified one novel restriction fragment length polymorphism in intron 15 of the gene. The frequency of the rare allele is: 0.08 in our control population. An increased frequency of this allele was found in CD patients but this difference did not reach statistical significance. A weak association between CD and homozygosity for the G allele of the IVS2+11A/G was found (OR [odds ratio] = 2.2, CI [confidence interval] = 1.3-3.9, chi2 = 8.4, p = 1.013). Nonparametric linkage analysis and transmissions disequilibrium tests did not provide evidence for linkage of NRAMP2 to IBD, UC, or CD. Sequence analysis of the exons and the iron-responsive element in a panel of 33 CD patients did not reveal any mutations in NRAMP2. Our association, linkage, and sequence analysis in IBD shows that the putative genetic risk factor on chromosome 12 likely is not NRAMP2. The weak association between the G/G genotype of IVS2+11A/G and CD may be due to linkage disequilibrium with a nearby disease causing gene. PMID- 10833068 TI - Evaluation of IBD candidate genes and the case of NRAMP2. PMID- 10833069 TI - Absence of GNAI2 codon 179 oncogene mutations in inflammatory bowel disease. AB - The human GNAI2 gene coding for G protein, Galphai2, is located on chromosome 3p21 in proximity to the region where an inflammatory bowel disease (IBD) locus has been suggested. Galphai2-deficient mice develop a lethal diffuse colitis that resembles human ulcerative colitis (UC) and frequently progresses to colon adenocarcinoma. Furthermore, the human GNAI2 gene is subject to point mutations at certain positions, including three at codon 179, all of which have been reported in human endocrine tumors. In order to evaluate the possible involvement of this gene in IBD pathogenesis, we have examined GNAI2 codon 179 sequences in 28 familial IBD patients, including 13 UC, 15 Crohn's disease (CD), and 7 patients with colon cancer/dysplasia, from 12 multiplex IBD families. The wildtype codon 179, CGC for arginine, plus the first G of the codon 180 engender a sequence recognizable by the enzyme BstUI. Mutations, therefore, can result in the abrogation of BstUI digestion of polymerase chain reaction (PCR) products containing the codon 179. Using the PCR-restriction fragment length polymorphism technique, all 28 IBD patients, including those with colon cancer, and 14 non-IBD family members show a BstUI-cleavable PCR-banding pattern indicating the presence of wildtype codon 179. We conclude that, in the familial IBD and colon cancer/dysplasia patients studied, there is no detectable mutation in the codon 179 of the GNAI2 gene. PMID- 10833070 TI - Probiotics and inflammatory bowel disease: is there a scientific rationale? AB - Most conventional forms of drug therapy suppress or modify the host immunoinflammatory response and neglect the other contributor to disease pathogenesis-the environmental microflora. Probiotics are live microbial food ingredients that alter the enteric microflora and have a beneficial effect on health. The rationale for using probiotics in IBD is mainly based on evidence from human studies and experimental animal models implicating intestinal bacteria in the pathogenesis of these disorders. The relationship between bacteria and intestinal inflammation is complex and does not appear to reflect a simple cause and effect. Similarly, the field of probiotics is complex and in need of rigorous research. Until the indigenous flora are better characterized and mechanisms of probiotic action defined, the promise of probiotics in IBD is unlikely to be fulfilled. Because of strain-specific variability and clinical and therapeutic heterogeneity within Crohn's disease and ulcerative colitis, it cannot be assumed that a given probiotic is equally suitable for all individuals. Although preliminary results of probiotic therapy in animal models and humans with ulcerative colitis and pouchitis have been encouraging, their efficacy in treatment or maintenance of remission of Crohn's disease remains to be clarified. However, the circumstantial evidence for some form of biotherapeutic modification of the enteric flora in Crohn's disease seems compelling. In the future, probiotics may offer a simple adjunct to conventional therapy with the emphasis on diet shifting from one of nutritional replenishment alone to a more functional role. PMID- 10833071 TI - Bacterial regulation of intestinal immune responses. AB - If we understand pathological processess within the alimentary tract, it is apparent that the fundamental aspects of microbe-host interactions need to be examined in greater detail. Pathogenic bacteria have evolved strategies to alter and subvert the function of T cels and phagocytes in the gut wall, and exploiting these molecules may lead to new treatments for chronic inflammatory bowel diseases. The adaptation of microbes to their host must involve microbe-mediated interference of the host innate immune response. The recent demonstration that nonpathogenic E. coli have a beneficial effect in ulcerative colitis further supports the notion that normal flora may alter the expression of the innate immune receptors or recognize alternative receptors compared with pathogenic variants. Such differences may conceivably lead to beneficial and protective alterations to the host through cytokine and antimicrobial peptide expression. Perhaps the contact point between microbes and host cells lies with the pattern recognition receptors such as TLRs. However, although much light has been shed on the downstream consequences of TLR activation, many more questions remain unsolved. For example, little is known about the expression profiles of the different TLRs throughout the gastrointestinal tract. Additionally, ambiguities remain over the natural ligands for TLRs. The discovery that the Drosophila Toll receptor acts downstream of the pathogen recognition event suggests that there are many more twists and turns to be revealed in the story of host-microbe interactions in the gastrointestinal tract. PMID- 10833072 TI - Prediction of treatment refractoriness in ulcerative colitis and Crohn's disease- do we have reliable markers? AB - Treatment refractoriness is a severe problem in the management of patients with ulcerative colitis and Crohn's disease. Despite some promising new therapeutic approaches, corticosteroids are still the preferential primary treatment for moderate to severe Crohn's disease and of severe ulcerative colitis. However, clinical response to corticosteroids varies, and many patients are resistant to such treatment. Since corticosteroids have frequent and even severe side effects, and toxicity increases with chronic steroid intake, factors predictive of response to such treatment would be very helpful for decisions on further management of these patients. At least in severe attacks of ulcerative colitis, the consensus seems to be that a high frequency of bowel movements as well as a high C-reactive protein and low serum albumin recorded after a few days of intensive medical treatment are important signs for early prediction of treatment failure in the majority of the patients. In Crohn's disease thus far, data on predictive factors are conflicting. No reliable marker with sufficient predictive value for treatment refractoriness could be identified. This might be due to the tremendous heterogeneity of Crohn's disease with many clinical phenotypes, which requires subgroup analysis with sufficient numbers of patients. Corticosteroids as well as other immunomodulating and immunosuppressive medications interfere with the immune system, which plays a central role in the mediation of intestinal inflammation. Treatment refractoriness might have its origin in specific immunological peculiarities eventually reflected in abnormal immunological, biochemical, and clinical parameters. Further exploration of those parameters to predict treatment refractoriness in patients with ulcerative colitis or Crohn's disease is of great clinical importance for safe and efficient management of patients. PMID- 10833073 TI - Infliximab is the drug we have been waiting for in Crohn's disease. PMID- 10833074 TI - Anti-TNF therapy for Crohn's disease: a perspective (infliximab is not the drug we have been waiting for) PMID- 10833075 TI - Anemia in IBD: the overlooked villain. AB - During the past decade relevant progress has been made in the understanding and treatment of IBD-associated anemia. Effective replacement of iron deficits has become safe by using novel intravenous iron preparations such as iron sucrose. The ability of erythropoietin to interfere with key mechanisms of myelosuppression in anemia of chronic diseases also benefits patients with IBD associated anemia. Concerns about cost effectiveness have been raised and weighed against the potential improvement in quality of life. Gastroenterologists who are caring for IBD patients should be concerned with low hemoglobin levels, since the quality of life in these patients can be as low as in anemic patients with advanced cancer. Also provided is a structured approach to cost-effective therapy. PMID- 10833076 TI - Thalidomide for Crohn's disease: high dose, low dose, or "no doze" at all? PMID- 10833077 TI - Reduction of stress--a new therapeutic goal in the treatment of inflammatory bowel disease? PMID- 10833078 TI - Budesonide: not for the long haul. PMID- 10833079 TI - Crohn's mucosa harbors adherent-invasive E. coli. PMID- 10833080 TI - Repetitive intradermal capsaicin: differential effect on pain and areas of allodynia and punctate hyperalgesia. AB - This study examined the effect of repeated intradermal capsaicin injections on capsaicin pain intensity and areas of allodynia and punctate hyperalgesia. Seventeen healthy volunteers participated in four sessions separated by at least 5 days. Each session included four intradermal injections of 10 microg of capsaicin. In one session injections were given with 0.5 cm and 6 min intervals, in a second with 0.5 cm and 15 min intervals, in a third with 0.5 cm and 24 min intervals, and in a fourth session with 4 cm and 15 min intervals. Following each injection capsaicin pain intensity was measured in the first 5 min, the area of allodynia at 5 min and area of punctate hyperalgesia at 10 min. With 6 min and 0.5 cm between repeated injections, capsaicin pain intensity decreased significantly whereas areas of allodynia and punctate hyperalgesia increased. In contrast, both capsaicin pain intensity and areas of allodynia and punctate hyperalgesia increased when the interval between injections was 24 min and 0.5 cm or 15 min and 4 cm. With 15 min and 0.5 cm between injections, capsaicin pain intensity did not change, whereas areas of allodynia and punctate hyperalgesia increased. There were no significant relations between capsaicin pain intensity and areas of allodynia and punctate hyperalgesia after first injections. The findings indicate that the response to intradermal injection of capsaicin is dependent on the time and distance between injections. The lack of significant relation between capsaicin pain intensity and area of allodynia and punctate hyperalgesia suggests that the two phenomena are mediated by different central mechanisms. PMID- 10833081 TI - Stability of rapidly adapting afferent entrainment vs responsivity. AB - Spike discharge activity was recorded from low-threshold, rapidly adapting, skin mechanoreceptive afferents (RA afferents) dissected from the median (forelimb) or tibial (hindlimb) nerves in anesthetized monkeys and cats. The spike activity was evoked by delivery of controlled sinusoidal vertical skin displacement ("flutter") stimuli to the receptive field (RF). The stimuli (15-30 Hz; 30-400 microm peak-to-peak amplitude; duration 0.8-15 s) were superimposed on a static skin indentation (0.5-1.0 mm) which was either maintained continuously throughout the run or applied trial-by-trial. The neural activity and the analog signal of the position of the stimulator probe were digitized at 10 kHz resolution and stored for off-line analysis. The main goal was to determine whether changes in the RA afferent response to skin flutter stimulation may be responsible for the enhanced capacity to discriminate stimulus frequency that accompanies a relatively brief (approximately 1 min) pre-exposure to such stimulation in humans. To this end, the spike train data were evaluated using methods that enabled independent measurement of entrainment and responsivity. Responsivity (response intensity) was measured as the average number of spikes/stimulus cycle, while entrainment (the degree to which evoked spike train activity is phase locked to the stimulus) was quantitatively assessed using statistical techniques developed for the analysis of "circular" (directional) data, supplemented by methods based on the calculation of power spectra from point process data. The methods are demonstrated to enable quantification of RA afferent entrainment over a range of stimulus durations and amplitudes substantially greater than reported in previous studies. While RA afferent responsivity was found to decline to a minor extent (10-20%) both across and within stimulus trials, entrainment remained consistently high and stable, and exhibited no temporal trends or dependence on any other measured factor. The average phase angle of the entrained RA afferent response also remained stable both within and across trials, showing only a tendency to increase slightly during the initial 100-500 ms after stimulus onset. The results imply that the improved capacity to discriminate stimulus frequency that develops in response to an exposure to cutaneous flutter stimulation is not attributable to a change in RA afferent entrainment per se. PMID- 10833082 TI - Contribution of central sensitization to the pain-related abnormal activity in neuropathic rats. AB - The possibility of different contributions from peripheral and central sensitization to distinct neuropathic pain syndromes has been studied in rats with chronic constriction of the sciatic nerve (CCI), showing positive behavioral signs of neuropathic pain. In anesthetized, paralyzed rats extracellular recordings were performed in the spinal sciatic afferent territory (L5-L6), ipsilateral to the injured nerve, from wide dynamic range (WDR) neurons. The spontaneous activity and the responses to noxious stimuli applied to the proper area, i.e., the skin innervated by the constricted sciatic nerve, and to "inappropriate" areas, like the tail and the area of skin supplied by the contralateral sciatic and saphenous nerves, were analyzed before and after input from the constricted nerve was reversibly blocked at the ganglionic level by local anesthetic. The neurons discharged spontaneously with high frequencies, and responded to the stimulation of proper and "inappropriate" areas with high frequency discharge and prolonged afterdischarges During the ganglionic block, confirmed by the lack of responses to proper area stimulation, theWDR neuron background activity was significantly reduced; the responses to all "inappropriate" afferences were present, the frequency discharges being comparable to the preblock ones while the afterdischarges were significantly shorter. Since the efficacy of "inappropriate" inputs is related to neuronal sensitization, the persistence of these responses indicates that central neurons remain sensitized during peripheral block. In view of the relationship between the examined spontaneous and stimulated activities and neuropathic pain symptoms, the data suggest that central sensitization contributes with different drive strength to such symptoms, playing a crucial role in extraterritorial pain. PMID- 10833083 TI - The organization of somatosensory cortex in the short-tailed opossum (Monodelphis domestica). AB - The organization of neocortex in the short-tailed opossum (Monodelphis domestica) was explored with multiunit microelectrode recordings from middle layers of cortex. Microelectrode maps were subsequently related to the chemoarchitecture of flattened cortical preparations, sectioned parallel to the cortical surface and processed for either cytochrome oxidase (CO) or NADPH-diaphorase (NADPHd) histochemistry. The recordings revealed the presence of at least two systematic representations of the contralateral body surface located in a continuous strip of cortex running from the rhinal sulcus to the medial wall. The primary somatosensory area (S1) was located medially while secondary somatosensory cortex (S2) formed a laterally located mirror image of S1. Auditory cortex was located in lateral cortex at the caudal border of S2, and some electrode penetrations in this area responded to both auditory and somatosensory stimulation. Auditory cortex was outlined by a dark oval visible in flattened brain sections. A large primary visual cortex (V1) was located at the caudal pole of cortex, and also consistently corresponded to a large chemoarchitecturally visible oval. Cortex just rostral and lateral to V1 responded to visual stimulation, while bimodal auditory/visual responses were obtained in an area between V1 and somatosensory cortex. The results are compared with brain organization in other marsupials and with placentals and the evolution of cortical areas in mammals is discussed. PMID- 10833084 TI - Time course of expression and function of the serotonin transporter in the neonatal rat's primary somatosensory cortex. AB - Immunocytochemical and autoradiographic techniques were employed to determine the time course of expression of the serotonin (5-HT) transporter (SERT) on thalamocortical afferents in the rat's primary somatosensory cortex (S-I), and to correlate this expression to the transient vibrissae-related patterning of 5-HT immunostaining previously described. In additional in vivo and in vitro experiments, 5-HT and 3H-5-HT were applied directly to the cortices of untreated and 5,7-dihydroxytryptamine-treated (5,7-DHT) rats in order to determine the period during which SERT functions on thalamocortical axons to take up 5-HT. In postnatal rats, SERT immunohistochemistry revealed a somatotopic patterning in S I that persisted until P-15, which is 6 days after the disappearance of the vibrissae-related 5-HT immunostaining. 3H-citalopram autoradiography revealed a vibrissae-related pattern in layer IV of S-I until at least P-30. Following destruction of raphe-cortical afferents with 5,7-DHT on the day of birth, this binding pattern remained visible until at least P-25, indicating that SERT located on thalamocortical axons is responsible for the 3H-citalopram patterning observed in S-I. Tissue from 5,7-DHT-treated rats that had 5-HT applied directly to their cortices revealed a normal vibrissae-related pattern of 5-HT immunostaining in S-I at P-7 and P-11 but only a faint pattern at P-13 and none at P-14. In addition, 3H-5-HT injected directly into S-I labeled layer IV barrels at P-6 and P-12 but not at P-18. The results of these experiments demonstrate that SERT is expressed by thalamocortical afferents and remains functional long after the vibrissae-related 5-HT immunostaining in cortex disappears. PMID- 10833085 TI - Assessment of spatial acuity at the fingertip with grating (JVP) domes: validity for use in an elderly population. AB - JVP domes are of a set of small grating surfaces recently introduced for cutaneous spatial resolution measurement. The gratings are placed on the skin and subjects are required to identify the orientation of grooves and bars. The finest grating whose orientations are discriminated reliably (75% correct) provides an estimate of the spatial resolution limit in the tested area. In the present study, we sought to determine the capacity of elderly subjects to resolve such grating stimuli in order to obtain normative data for this population. Thirty-two elderly individuals in good health (range: 60-88 years) were assessed for their ability to perceive grating orientation at the tip of the dominant index finger. Testing proceeded from the widest grating dome (3 mm) to the next (e.g., 2 mm), until the performance level dropped below 75% correct discrimination. The grating orientation task proved to be very difficult for most subjects and only a minority (14/32) was able to provide reliable reports of grating orientation even with presentation of the widest dome available (3 mm). Accordingly, individual grating resolution thresholds were often considerably higher (> 2.5 mm, n = 26) than values previously reported in young adults for the fingertip region (approximately 1 mm). These results suggest that the current set of grating domes may not be adequate for spatial acuity measurement at the fingertip of older adults. New larger grating dimensions should be added to the set presently available to improve their sensitivity for an older population. PMID- 10833086 TI - Characteristics of GABAergic neurons and their synaptic relationships with intrinsic axons in the cat motor cortex. AB - The intrinsic circuitry of the motor cortex comprises a complex network of connections whose synaptic relationships are poorly understood. This study was designed to determine the characteristics of subsets of GABAergic neurons containing the calcium-binding proteins parvalbumin (PV) and calbindin (CB), and their relationships with intrinsic axons in motor cortex. Immunohistochemically identified PV-containing neuronal profiles were more evenly distributed across cortical laminae (38% in II-III, 32% inV, 30% in VI) and more numerous (2.1/1) than CB-containing neuronal profiles (71% in II-III, 17% in V, 12% in VI). Relationships between neurons and axons intrinsic to motor cortex were visualized with fluorescent markers using the laser scanning confocal microscope. Similar percentages of PV (43%) and CB-immunoreactive (IR) (40%) neurons formed sparsely distributed appositions (1-5/neuron) with anterogradely labeled axons. The mean distances of such appositions from the somata were significantly different for the two groups (PV, mean = 22 microm, range = 1.6-93 microm; CB, mean = 32 microm, range = 6.2-132 microm). PV-IR neurons had a lower ratio of axosomatic/axodendritic appositions (1/99) compared with CB-IR neurons (14/86). Ultrastructural studies confirmed these findings. Fifty-seven percent of CB-IR neurons and 38% of PV-IR neurons formed synapses with intrinsic axons. Both populations received sparse input (1-6 synapses/neuron). Nearly all appositions between labeled terminals and postsynaptic profiles formed one synapse. Postsynaptic dendrites of PV-IR neurons (mean = 1.4 microm diameter) were larger than those of CB-IR neurons (mean = 1.1 microm), indicating more proximal synapses. Distinct input patterns of intrinsic axons to the two populations of neurons suggest unique roles in cortical processing. PMID- 10833087 TI - Barrels XII--proceedings. 22-23 October 1999, Miami Lakes, Florida, USA. PMID- 10833088 TI - How come I've got it? (A review of Helicobacter pylori transmission). AB - Numerous epidemiological studies have shown the two main risk factors for Helicobacter pylori acquisition: childhood and low socio-economic level both in developing and developed countries. Nevertheless, in the absence of ubiquitous extra-human reservoir(s), the route(s) for person-to-person transmission remains undetermined. Very recent data favour the oro-oral route: besides gastric mucosa, mouth might be a sanctuary site and the oro-oral transmission hypothesis seems applicable worldwide. Nevertheless, the gastro-oral route (vomiting, gastro oesophageal reflux) is still possible and deserves further research. In developing countries, feco-oral transmission (perhaps through the water supply) might be a significant mechanism of human contamination both for primary infection in children and perhaps, reinfection in adults. PMID- 10833089 TI - Prevalence of Helicobacter pylori infection in Kenyan schoolchildren aged 3-15 years and risk factors for infection. AB - OBJECTIVES: To determine the prevalence of Helicobacter pylori (Hp) infection in Kenyan schoolchildren aged 3-15 years and the risk factors for infection. DESIGN: A cross-sectional survey and study of risk factors. SUBJECTS: Primary-school children in Trans Nzoia district, Kenya. METHODS: Hp status was determined using the 13C-urea breath test. Risk factors were assessed using a structured interview schedule. RESULTS: Of 205 children interviewed, valid breath test results were obtained for 192 (93.7%), of whom 155 (80.7% (95% CI 75.2-86.3)) were infected. Univariate analysis showed weak associations between Hp infection and rural areas (location of home, Relative Risk (RR)= 1.2 (1.0-1.4); location of school, RR = 1.2 (1.0-1.3)) and lower socio-economic status (RR = 1.2 (1.0-1.4)). Access to only a pit-latrine in the home (RR = 1.2 (1.0-1.4)) and plate-sharing with other family members in the home also increased the likelihood of Hp infection (RR = 1.3 (1.1 -1.6)). However, after logistic regression modelling only plate-sharing in the home remained as an independent risk factor (odds ratio (OR) = 2.8 (1.3 6.3)). CONCLUSIONS: The prevalence of Hp infection in Kenyan schoolchildren aged 3-15 years is already high, suggesting that most acquisition occurs before age 3 years. Children who normally shared a plate with other family members during meals in the home had a higher prevalence of infection, suggesting that oro-oral transmission may be important in this population. Future studies in developing countries should focus on younger children (before age 3 years), and consider the possibility of adult-child, oro-oral transmission in order to establish likely modes of transmission. PMID- 10833090 TI - One-week therapy with pantoprazole versus ranitidine bismuth citrate plus two antibiotics for Helicobacter pylori eradication. AB - AIM: A combination of omeprazole plus amoxycillin (Amo) and clarithromycin (CIa) for 7 days has been studied extensively. However, the role of other proton pump inhibitors, such as pantoprazole (Pan), in this therapy is not well known. On the other hand, ranitidine bismuth citrate (RBC) also seems to be effective when combined with Amo and CIa. Our aim was to evaluate and to compare these two novel short-term triple therapies (Pan+Amo+Cla and RBC+Amo+Cla) for treatment of Helicobacter pylori. METHODS: In a randomized clinical trial 150 consecutive patients (38 with duodenal ulcer, 112 with non-ulcer dyspepsia) infected by H. pylori were studied prospectively. Exclusion criteria were: previous H. pylori eradication therapy, gastroerosive drug use, gastric surgery, and associated diseases. One of two regimens was given for 7 days: Pan (40 mg b.i.d.), Amo (1 g b.i.d.), Cla (500 mg b.i.d.) (group Pan+Amo+Cla, n = 75); or RBC (400 mg b.i.d.), Amo (1 g b.i.d.), Cla (500 mg b.i.d.) (group RBC+Amo+Cla, n = 75). All drugs were administered together after meals. Compliance was evaluated by return tablet count. Data were analysed by univariate (chi2) and multivariate (multiple logistic regression) analysis. Eradication was defined as a negative 13C-urea breath test 1 month after completing therapy. RESULTS: The distribution of studied variables (age, gender, smoking, duodenal ulcer/non-ulcer dyspepsia) was similar in both therapy groups. Per-protocol eradication was achieved in 48/71 (68%) in group Pan+Amo+Cla, and in 61/70 (87%) in group RBC+Amo+Cla (P= 0.01). Intention-to-treat (ITT) eradication was achieved in, respectively, 48/ 75 (64%) and in 61/75 (81%) (P= 0.03). The RBC+ Amo+Cla regimen was more effective than Pan+Amo+Cla in non-ulcer dyspepsia patients (ITT, 84% vs 58%; P = 0.005), but statistically significant differences were not demonstrated in duodenal ulcer patients (72% vs 80%). In the multivariate analysis the odds ratio for the effect of the type of therapy on H. pylori eradication in patients with non-ulcer dyspepsia was 3.8 (95% Cl, 1.6-9.3; P = 0.003). No relevant adverse effects were reported with any regimen. CONCLUSION: A RBC+Amo+Cla regimen for only 1 week is a promising therapy for H. pylori infection, due to its high efficacy, simple posology, and excellent tolerability. Combination of Pan with Amo and Cla, although effective in duodenal ulcer patients, but in non-ulcer dyspepsia has not achieved the favourable results previously reported with other proton pump inhibitors. PMID- 10833091 TI - Gastric alpha-tocopherol and beta-carotene concentrations in association with Helicobacter pylori infection. AB - OBJECTIVE: The effects of Helicobacter pylori infection and its associated gastric histology on alpha-tocopherol and beta-carotene concentrations in serum, gastric juice and antral mucosa were investigated in patients undergoing routine gastroscopy for investigation of dyspepsia. METHOD: Eighty-six patients were studied. High-performance liquid chromatography was used to measure alpha tocopherol and beta-carotene concentrations. H. pylori infection was assessed by histology, bacterial culture, rapid urease test and serology. RESULTS: No obvious association was found between age, sex, smoking or endoscopic diagnosis and alpha tocopherol or beta-carotene concentrations in serum, gastric juice and antral mucosa. However, alcohol drinkers had significantly lower antral mucosal and gastric juice beta-carotene concentrations compared to non-drinkers. Gastric juice beta-carotene concentration was markedly lower in patients infected with H. pylori than uninfected controls (2.9 nmol/l (interquartile range 0.3-4.3) versus 4.6 nmol/l (interquartile range 3.5-7.6), P = 0.01), but there was no significant difference in serum or gastric mucosal beta-carotene concentrations between the two patient groups. The presence of gastric atrophy and intestinal metaplasia was significantly associated with reduced mucosal alpha-tocopherol and beta-carotene concentrations. Furthermore, antral mucosal alpha-tocopherol concentrations decreased progressively as antral mucosal histology changed from normal to chronic gastritis alone and finally to atrophy and intestinal metaplasia. CONCLUSION: Gastric alpha-tocopherol and beta-carotene concentrations are affected by H. pylori-associated gastric histological changes, and these findings suggest that H. pylori infection may not only impair the protective role of vitamin C, but also of alpha-tocopherol and beta-carotene in the stomach. PMID- 10833092 TI - Depression during interferon therapy for chronic viral hepatitis: early identification of patients at risk by means of a computerized test. AB - OBJECTIVES: At the doses used for the treatment of chronic viral hepatitis, interferon (IFN)-related side-effects are usually modest, even though in some cases they require the interruption of therapy. Neuropsychiatric disturbances that range from modest depression and irritability to forms of manic-depressive psychosis and attempted or successful suicides are among the most important side effects. The aim of our study was to determine whether the Minnesota Multiphasic Personality Inventory (MMPI) is a sensitive and reliable test for the early identification of patients at risk of depression before IFN therapy is commenced, and whether it could be useful for the monitoring of these patients during treatment. METHODS: We prospectively studied 67 patients with chronic active liver diseases, consecutively enrolled in open studies and treated with r IFNalpha2. Before starting therapy and after 3 months of treatment, all patients underwent a clinical neurological evaluation and MMPI. RESULTS: At baseline, the correlation between the clinical evaluation and the score of the depression scale of the MMPI was statistically significant (P< 0.0001). Nine of 14 (64.3%) patients with a baseline score > or = 60/100 showed a depressive mood after 3 months of therapy. Five of 44 patients (11.3%) with a baseline score < 60/100 showed a depression of medium level after 3 months of treatment. This difference was highly significant (P< 0.0001). CONCLUSIONS: According to our results, the MMPI is a reliable and sensitive test for the early identification of patients at risk of depression before and during IFN therapy for chronic viral liver diseases. PMID- 10833093 TI - Efficacy of ursodeoxycholic acid in association with alpha-interferon for chronic hepatitis C in alpha-interferon non-responder patients. AB - BACKGROUND: The aim of the present study was to evaluate the effect of combined treatment with alpha-interferon (alpha-IFN) and ursodeoxycholic acid (UDCA) on liver function tests and serum HCV-RNA in patients with chronic hepatitis C who had not responded to alpha-IFN alone. METHOD: One hundred and three patients (60 men, 43 women, mean age 49 +/- 1.3 years) who had not responded (both HCV-RNA positive and increased serum ALT levels) to 4 consecutive months of treatment with alpha-IFN (3 MU three times weekly) were randomly assigned to receive UDCA (IFN-UDCA, 53 patients, 600 mg/day) in addition to the same alpha-IFN dose, or to continue alpha-IFN alone (IFN-controls, 50 patients). After stopping alpha-IFN, patients who had received UDCA continued to receive UDCA for an additional 6 month period. The two groups were comparable for sex, basal ALT, basal yGT, genotype distribution and liver histology, while mean age was lower in controls (53 +/- 1.8 vs 46 +/- 1.8 years; P< 0.01). RESULTS: Twenty (38%) out of 53 IFN UDCA patients had normal ALT, compared with only six (12%) out of 50 IFN-control patients (P < 0.01). HCV-RNA became undetectable in four IFN-UDCA patients. Three months after withdrawal of alpha IFN, 15 IFN-UDCA responders, but none of the IFN controls, had normal ALT values (P< 0.01); 6 months after withdrawal, nine IFN UDCA responders still had normal ALT (P= NS) and HCV-RNA was still undetectable in four of them. Portal and periportal inflammation showed a statistically significant improvement (Fisher's exact test P< 0.01) in IFN-UDCA patients as compared with IFN-controls, while no effect was observed on portal fibrosis. CONCLUSIONS: These data demonstrate that UDCA improves the response rate to alpha IFN. Furthermore, in 8% of IFN-UDCA patients the response rate was sustained and associated with HCV-RNA clearance. PMID- 10833094 TI - Short-term prognosis in critically ill patients with liver cirrhosis: an evaluation of a new scoring system. AB - OBJECTIVE: The mortality of patients with liver cirrhosis admitted to an intensive care unit (ICU) has been found to be high. This study was performed to assess the physiological and laboratory parameters which are able to identify on ICU admission the cirrhotic patients who are most likely to die. DESIGN: Prospective clinical trial. METHODS: Two groups of patients were analysed. Group A consisted of 196 consecutive cirrhotic patients admitted to our medical ICU for various reasons. For the detection of independent outcome predictors, we used a multiple logistic regression model. Based on these variables, the 'intensive care cirrhosis outcome (ICCO) score' was calculated. The ability to discriminate between survivors and non-survivors was determined by receiver operating characteristic curves, and the area under the curve was calculated. Group B consisted of 70 consecutive cirrhotic patients for prospective validation of the ICCO score. RESULTS: Applying multiple logistic regression analysis, bilirubin, cholesterol, creatinine clearance and lactate were found to be independently associated with the hospital mortality. The ICCO score was 0.3707 + (0.0773 x bilirubin (mg/dl)) - (0.00849 x cholesterol (mg/dl)) -(0.0155 x creatinine clearance (ml/min)) + (0.1351 x lactate (mmol/l)), giving an area under a receiver operating characteristic curve of 0.9. Increasing score values were associated with an increase in mortality. All patients with an ICCO score > +2.6 died. CONCLUSIONS: Application of the ICCO score is rapid and available at the patient's bedside, and its application is simple and reproducible. In cirrhotic patients, the ICCO score has a high ability to discriminate between survivors and non-survivors. The ICCO score may facilitate estimation on ICU admission of the prognosis of critically ill cirrhotic patients. PMID- 10833095 TI - A prospective series of out-patient endoscopic retrograde cholangiopancreatography. AB - OBJECTIVE: Out-patient endoscopic retrograde cholangiopancreatography (ERCP) is widely practised but the safety of this approach has only been addressed in a limited number of prospective series mainly from specialist North American centres. Our objective was to determine prospectively the safety and admission rates of out-patient ERCPs. STUDY DESIGN AND PARTICIPANTS: Patients were selected for out-patient ERCP if in relatively good health, without major risk factors for complications following ERCP and with adequate social support. Our series consisted of 136 consecutive cases of which 82 were therapeutic. SETTING AND OUTCOME MEASURES: A district general hospital in the UK, which also performs ERCPs for neighbouring health districts. Out-patient ERCP patients were followed up at 30 days using standard criteria for defining complications. RESULTS: Procedures were 60 biliary sphincterotomy, 10 stone removal, nine stenting procedures, two dilatations and one pancreatic intervention. Complications were pancreatitis in seven patients (six moderate severity, one mild), cholangitis in three patients, haemorrhage in one patient. Nine patients required admission for complications, two from the endoscopy unit and seven from home; their average in patient stay was 6 days. Seventeen patients were admitted for observation or for further management. There was one death unrelated to ERCP. Overall, 110 of 136 patients did not require inpatient care following out-patient ERCP. CONCLUSIONS: Our complication rates were similar to those of other series. Out-patient ERCP for selected cases, with adequate post-discharge arrangements for advice and readmission, appears safe and would reduce healthcare costs. PMID- 10833096 TI - Gastric permeability to sucrose is increased in portal hypertensive gastropathy. AB - BACKGROUND: Portal hypertensive gastropathy (PHG) is frequently found among patients with hepatic cirrhosis and at present the only way to detect and follow PHG is via endoscopy. OBJECTIVE: To assess gastric and intestinal permeability and investigate its relationship to endoscopic findings and indices of portal hypertension and hepatic function. DESIGN AND METHODS: Thirty-one non-diabetic patients with hepatic cirrhosis and PHG (PHG+) were studied and compared with 17 cirrhotic patients without PHG (PHG-). All patients underwent endoscopy for the assessment of PHG and Helicobacter pylori status, ultrasound determination of the diameters of spleen and portal vein, and, subsequently, an oral load of sucrose, lactulose, and mannitol. Sugar concentrations were determined in 6-h urine specimens and expressed as a percentage of the orally administered dose or as lactulose/mannitol ratio. RESULTS: The urinary sucrose excretion was significantly elevated in patients with PHG compared to those without (PHG+, 0.20% +/- 0.03; PHG-, 0.07% +/- 0.01; P< 0.001). No difference was found for the small intestinal probes lactulose and mannitol. Gastric sucrose permeability correlated positively with the endoscopic lesion score (P < 0.001), but not with other parameters of portal hypertension or hepatic function. H. pylori status did not influence gastric permeability. The sensitivity of this test reached 100% for PHG scores > 2. CONCLUSIONS: Gastric permeability to sucrose is increased in patients with PHG, independently of the presence of H. pylori. Sucrose permeability may be useful for the follow-up of patients with PHG. PMID- 10833097 TI - A randomized controlled trial of rubber band ligation versus infra-red coagulation in the treatment of internal haemorrhoids. AB - OBJECTIVE: Despite the presence of numerous non-surgical therapies for the treatment of haemorrhoids, none of these therapies has clearly been proven to be superior. The effectiveness and patient tolerance of rubber band ligation (RBL) and infra-red coagulation (IRC) in the treatment of haemorrhoids was assessed. DESIGN: Prospective randomized trial. SETTING: Academic hospital (tertiary care). PARTICIPANTS: A total of 133 consecutive patients (73 males, 60 females, mean age 48 years (range 19-82)) with internal haemorrhoids, and without concomitant anorectal disease, were randomized to rubber band ligation (RBL, n = 65) or infra red coagulation (IRC, n = 68). INTERVENTIONS: Rubber band ligation or infra-red coagulation was performed in one or more sessions with four-week intervals until symptoms had resolved. Treatment outcome and side-effects were assessed after each treatment session and one month after the last treatment by proctological examination and a questionnaire, including a pain score (visual analogue scale from 0 to 10). Recurrence of complaints was assessed by telephone questionnaire [mean follow-up of 19.2 months (SD 7.8)]. RESULTS: Treatment outcome was assessed in 124 patients (60 RBL, 64 IRC). The mean number of treatment sessions was 1.6 (SD 0.9) for both therapies. For RBL, 58 patients (97%), and for IRC, 59 patients (92%) were symptom-free or had satisfactorily improved. Only third-degree haemorrhoids seemed to respond better to RBL (five of five patients symptom-free) than to IRC (two of four patients symptom-free). Pain following treatment was more common and more severe after RBL (VAS 5.5 +/- 3.7) than after IRC (VAS 3.3 +/- 3.3, P= 0.018). The telephone questionnaire was answered by 105 patients. Nine of 50 patients (18%) treated with RBL and 11 of 55 patients (20%, P= 0.81) treated with IRC had experienced symptomatic relapse to pre-treatment levels. CONCLUSIONS: Infra-red coagulation and rubber band ligation are equally effective in the treatment of haemorrhoids. The rate and severity of pain is higher after rubber band ligation. Infra-red coagulation should be the first-line treatment for haemorrhoids. PMID- 10833098 TI - Impact of bile acid malabsorption on steatorrhoea and symptoms in patients with chronic diarrhoea. AB - OBJECTIVE: Bile acids are important for fat absorption. The relationship between bile acid malabsorption and steatorrhoea and gastrointestinal symptoms in patients with chronic diarrhoea has only been studied on a limited scale. DESIGN: Ninety-four patients referred for chronic diarrhoea were prospectively investigated with the 75SeHCAT test, a faecal fat excretion test and registration of symptoms in addition to the standard clinical work-up. METHODS: The correlation between the 75SeHCAT value and the faecal fat excretion was calculated for different groups of patients. Symptoms were registered in a questionnaire over a period of seven consecutive days. RESULTS: Forty-two patients had a 75SeHCAT value < 10%. Mild steatorrhoea was common in patients with non-organic bile acid malabsorption (50%) and in patients with functional diarrhoea (38%). There was no correlation between low 75SeHCAT values and steatorrhoea, although some patients with severe organic disease had a concomitant malabsorption of fat and of bile acids. In coeliac disease, severe steatorrhoea was common even in patients with high 75SeHCAT values. Patients with bile acid malabsorption had more frequent (P < 0.008) and looser (P= 0.0021) stools compared with patients with functional diarrhoea. There was no difference in abdominal pain, distension or flatulence. CONCLUSION: Mild steatorrhoea is common in both non-organic bile acid malabsorption and functional diarrhoea. The 75SeHCAT value cannot predict the risk of steatorrhoea. The high prevalence of bile acid malabsorption in patients with chronic diarrhoea and the absence of specific symptoms, except frequent and more liquid stools, indicates that the 75SeHCAT test should be performed early in the investigation of these patients. PMID- 10833099 TI - The detection of cytokeratins in lymph nodes of Duke's B colorectal cancer subjects predicts a poor outcome. AB - OBJECTIVES: The objectives of this study were to examine the frequency of lymph node micrometastases detected by keratin immunohistochemistry and their relationship with survival behaviour. METHODS: A total of 133 consecutive patients staged as Duke's B, who had curative resection for colorectal cancer (CRC), comprised the study population. Patients who had died of a non-CRC-related cause or who became lost to follow-up were excluded, resulting in an amended population of 100. Study end-points were defined as disease-free survival of 5 years or CRC-related death. Paraffin-embedded lymph node sections were stained with a commercial cytokeratin antibody using a standard avidin-biotin technique. RESULTS: One quarter of subjects had micrometastases. Fifty-six per cent of subjects with positive lymph nodes had an adverse outcome, compared with 11% of subjects with negative nodes. A highly significant association was found between lymph node cytokeratin expression and mortality in both the univariate (log rank P = 0.0001) and multivariate (Cox proportional hazards P = 0.0123) analysis. CONCLUSIONS: Lymph node micrometastases detected by this inexpensive and simple technique are significantly associated with mortality in Duke's B CRC. This technique may be used to select patients for adjuvant chemotherapy. PMID- 10833100 TI - Luminal neutrophil migration in ileoanal pouches studied by whole gut lavage. AB - OBJECTIVE: White cell scintigraphy has shown that neutrophils migrate into the gut wall and lumen in ileoanal pouches. We aimed to establish whether whole gut lavage fluid can be used to investigate intestinal neutrophil migration in ileoanal pouches. DESIGN: A prospective single centre study recruiting consecutive patients. METHODS: Whole gut lavage with polyethylene glycol electrolyte solution was performed in 56 (32 men, 24 women) ileoanal pouch patients who had undergone colectomy for ulcerative colitis; the first clear effluent was collected, processed and stored at -70 degrees C. The fluid was assayed for neutrophil granulocyte elastase using a specific colorimetric assay, IgG, albumin, alpha1-antitrypsin, haemoglobin and cytokines IL-1beta and IL-8 using previously described techniques. Patients' disease activity was characterized following pouchoscopy and biopsy. RESULTS: Patients with pouchitis had significantly higher levels of granulocyte elastase in whole gut lavage fluid compared with those without pouchitis. Patients with detectable granulocyte elastase had higher pouchoscopy score, more severe mucosal neutrophil infiltration and protein loss and bleeding. These patients had significantly higher levels of cytokines IL-1beta and IL-8 in the whole gut lavage fluid, compared with patients with undetectable granulocyte elastase. CONCLUSION: Whole gut lavage fluid samples may provide a useful investigative tool to study mucosal inflammation and luminal neutrophil migration in ileoanal pouches. PMID- 10833101 TI - Overlap syndrome between autoimmune hepatitis and primary sclerosing cholangitis in two cases. AB - The overlap syndrome between autoimmune hepatitis and primary sclerosing cholangitis is a rare condition and only few cases have been published, partly associated with ulcerative colitis, but not with Crohn's disease. We report an autoimmune hepatitis/primary sclerosing cholangitis overlap syndrome in a female patient with Crohn's disease. In addition, a second case of overlap syndrome is reported in a man without inflammatory bowel disease. A 24-year-old woman was referred with a 10-month history of diarrhoea and biochemical changes including elevated serum levels of alkaline phosphatase, aspartate aminotransferase, alanine aminotransferase and immunoglobulin G. Enzyme linked immunosorbent assay showed that antinuclear autoantibodies were elevated. Immunofluorescence for perinuclear-staining antineutrophil cytoplasmatic antibodies was positive. Diagnostic criteria of definite autoimmune hepatitis according to the International Autoimmune Hepatitis Group were fulfilled. Liver biopsy simultaneously showed criteria of autoimmune hepatitis and primary sclerosing cholangitis. Endoscopic retrograde cholangiography demonstrated features of primary sclerosing cholangitis. Colonoscopy and colonoscopic biopsies indicated an active Crohn's disease affecting the terminal ileum and the ascending and transverse colon. Furthermore, we report the case of a 28-year-old man with known primary sclerosing cholangitis for the previous 6 years, and who developed jaundice and a marked increase of aspartate aminotransferase, alanine aminotransferase and immunoglobulin G, leading to the diagnosis of definite autoimmune hepatitis. A review of the literature revealed only 16 cases of an autoimmune hepatitis/primary sclerosing cholangitis syndrome in patients without inflammatory bowel disease or in association with ulcerative colitis. We report two additional cases, one case showing an association with Crohn's disease. PMID- 10833102 TI - Simultaneous occurrence of focal nodular hyperplasia and primary sclerosing cholangitis in a young female. AB - We report the case of a 22-year-old female with primary sclerosing cholangitis who was found, during hepatic imaging, to have a large liver mass. Imaging techniques and histological examination confirmed the mass to be focal nodular hyperplasia. A review of the literature indicates that the simultaneous occurrence of these two hepatic pathologies is unique. The differential diagnosis of hepatic masses in primary sclerosing cholangitis is discussed. Focal nodular hyperplasia needs to be included in the differential diagnosis of hepatic lesions in primary sclerosing cholangitis. PMID- 10833103 TI - Portal hypertension in the presence of minimal liver damage in Crohn's disease on long-term azathioprine: possible endothelial cell injury. AB - Azathioprine is a useful agent for the treatment of Crohn's disease but side effects occur in 10% of patients. Hepatic toxicity is well recognized and is usually associated with abnormalities of liver function tests. We describe a female patient who was on azathioprine for the treatment of Crohn's disease for a total of 216 months. She developed portal hypertension complicated by variceal haemorrhage. This required the insertion of a trans-jugular intra-hepatic porto systemic shunt to control the bleeding. Subsequent histology has shown mild nodular regenerative hyperplasia, and other causes of liver disease have been excluded. The only liver function abnormalities were mild elevation of bilirubin and a low albumin in the later stages. PMID- 10833104 TI - Fulminant hepatic failure as the initial manifestation of primary hepatocellular carcinoma. AB - A 20-year-old male, with no history of chronic liver disease, presented with fulminant hepatic failure. The cause was not established until post mortem when it was found that he had a primary hepatocellular carcinoma in a non-cirrhotic liver. To our knowledge, this is the third report of hepatocellular carcinoma presenting in this manner. Although rare, primary malignancies of the liver should be considered in the differential diagnosis of fulminant hepatic failure, especially when other more common causes are excluded. PMID- 10833105 TI - Late recurrence of a hepatic angiomyolipoma. AB - Angiomyolipomas are benign mesenchymal tumours, mostly of renal origin. Hepatic angiomyolipomas are rare, and radiological and pathological diagnoses may be difficult We report on the first case of hepatic angiomyolipoma recurrence known to us, 6 years after surgical treatment of the initial tumour. Moreover, this hepatic recurrence was associated with renal angiomyolipoma without any stigmata of tuberous sclerosis. PMID- 10833106 TI - Toxic megacolon: a rare presentation of primary lymphoma of the colon. AB - Primary colonic lymphoma is rare and accounts for less than 1% of colon malignancies. Moreover, diffuse neoplastic invasion of the colon is exceptional. This case describes a patient with primary non-Hodgkin's lymphoma of the colon presenting as toxic megacolon. This unique presentation is the first case reported in the literature of a diffuse colonic lymphoma diagnosed in the setting of a toxic megacolon. Histological confirmation was performed on the piece of colectomy. PMID- 10833107 TI - Mycophenolate mofetil for primary biliary cirrhosis--convergent thinking. PMID- 10833108 TI - Defining the role of helical computerised tomography (CT) in the diagnosis of pulmonary embolism (PE) PMID- 10833109 TI - Lung volume reduction surgery (LVRS) for emphysema--early Australian experience. PMID- 10833110 TI - Suspected acute pulmonary emboli: cost-effectiveness of chest helical computed tomography versus a standard diagnostic algorithm incorporating ventilation perfusion scintigraphy. AB - BACKGROUND: There is a controversy regarding the investigation of patients with suspected acute pulmonary embolism (PE). AIMS: To compare the cost-effectiveness of alternative methods of diagnosing acute PE. Chest helical computed tomography (CT) alone and in combination with venous ultrasound (US) of legs and pulmonary angiography (PA) were compared to a conventional algorithm using ventilation perfusion (V/Q) scintigraphy supplemented in selected cases by US and PA. METHODS: A decision-analytical model was constructed to model the costs and effects of the three diagnostic strategies in a hypothetical cohort of 1000 patients each. Transition probabilities were based on published data. Life years gained by each strategy were estimated from published mortality rates. Schedule fees were used to estimate costs. RESULTS: The V/Q protocol is both more expensive and more effective than CT alone resulting in 20.1 additional lives saved at a (discounted) cost of $940 per life year gained. An additional 2.5 lives can be saved if CT replaces V/Q scintigraphy in the diagnostic algorithm but at a cost of $23,905 per life year saved. CONCLUSIONS: The more effective diagnostic strategies are also more expensive. In patients with suspected PE, the incremental cost-effectiveness of the V/Q based strategy over CT alone is reasonable in comparison with other health interventions. The cost-effectiveness of the supplemented CT strategy is more questionable. PMID- 10833111 TI - Videoscopic lung volume reduction surgery in an Australian public teaching hospital. AB - BACKGROUND: Lung volume reduction surgery (LVRS) has been a frequent literature topic in emphysema management recently. Opinions differ in regard to usefulness, efficacy, and selection criteria. AIMS: To present the results of our first 55 bilateral videoscopically resected group, with follow-up of up to three years, and to present some of the local methodology problems faced. METHODS: Thirty-nine men and 16 women, age range 40-77, had either upper lobe (42), mixed (two), or lower lobe (11) resections without buttressing (except for unilateral buttressing in several of the latter patients as part of an intrapatient comparison trial) according to their pattern of emphysema determined by CT and perfusion scanning. RESULTS: Thirty day mortality was 5.5%. Follow-up pulmonary function is available for 44 patients, and demonstrates a mean 51% improvement in FEV1, and significant improvement in FVC, PaO2, dyspnoea indices and walking distance, with a reduction in mean RV, TLC, PaCO2. FEV1 improvement is maintained above baseline at three years. Lower lobe surgery outcomes are at least as good as their upper lobe counterparts. CONCLUSIONS: Outcomes confirm improvements reported elsewhere, and suggest that videoscopic resection may provide worthwhile benefit to lower lobe patterns of emphysema. Other managment issues are discussed. PMID- 10833112 TI - Vitamin D deficiency and secondary hyperparathyroidism: clinical and biochemical associations in older non-institutionalised Southern Tasmanians. AB - AIM: To determine the prevalence and associations of vitamin D (25-OHD) deficiency in a sample of older Tasmanian subjects. METHODS: A cross-sectional survey of: 109 patients with a mean age of 79 years (range 60-101 years) consecutively admitted to a short stay geriatric rehabilitation ward; 52 community dwelling subjects with a mean age of 75 years (range 64-88 years). Subjects answered a questionnaire, had anthropometric measurements and underwent venepuncture. RESULTS: The main outcome measure was 25 hydroxy vitamin D (25-OHD) level with deficiency defined as <28 nmol/L. Vitamin D deficiency was found in 67% and secondary hyperparathyroidism in 49% of the hospitalised group. Vitamin D deficiency was also found in 17% of the community group, in particular one in three residents of Independent Living Units was deficient. Subjects who were deficient were older (80 years vs 76 years [p<0.001]), had lower body mass index (23.7 kg/m2 vs 25.9 kg/m2 [p<0.001]) and had a lower serum albumin (35 gm/L vs 39 gm/L [p<0.001]). Deficient subjects had poorer physical functional status (p=0.02) and lower activity levels (p<0.001) and reported less habitual sun exposure (p<0.001). Biochemical measures such as parathyroid hormone, alkaline phosphatase and calcium were weakly predictive of vitamin D levels. By stepwise multiple regression analysis, the only significant predictors of vitamin D levels were the Frenchay Activity Index, albumin and calcium. CONCLUSION: Vitamin D deficiency and secondary hyperparathyroidism is common in community living older people who are hospitalised in Southern Tasmania and is associated with increasing age, poor physical function and activity and low reported sun exposure. PMID- 10833113 TI - Male systemic sclerosis and occupational silica exposure-a population-based study. AB - BACKGROUND: The continuing uncertainty about the silica-systemic sclerosis relationship led to the investigation of its role as a disease determinant in a large population-based study of systemic sclerosis. AIMS: To compare the frequency, socioeconomic and educational status, age-specific prevalence and duration of occupational silica exposure in males with and without systemic sclerosis. To assess the temporal relationship between exposure and disease onset. To estimate disease latency. To compare disease characteristics between silica-exposed and non-silica-exposed male cases. METHODS: The study was case control in design. The exposure variable was occupational silica exposure as assessed by an occupational health officer blinded to case/control status and the outcome variable was systemic sclerosis. The employed instrument comprised either a standardised telephone questionnaire (interviewed cases and controls) or medical records (deceased or living-status-unknown cases). RESULTS: Sixty of 160 cases (37.5%) and 11 of 83 (13.3%) controls had occupational silica exposure (OR=3.93; 1.84-8.54). Comparison of data between 64 interviewed cases and all controls demonstrated initial occupational silica exposure occurring before age 40, comparable educational status but significantly different cumulative socioeconomic status with cases being over-represented in semi-skilled and unskilled occupations. Cross-sectional 'current' occupational data underestimated cumulative silica exposure by more than 50%. Silica exposure uniformly preceded onset of second disease symptoms and disease diagnosis. In most, it also preceded onset of first disease symptoms. Disease latency approximated two decades. No disease features distinguished silica-associated systemic sclerosis from idiopathic systemic sclerosis. The duration of silica exposure in the interviewed silica-exposed cases did not significantly exceed that of silica-exposed controls. CONCLUSIONS: Male systemic sclerosis displays socioeconomic dependence. Silica is a disease determinant in male systemic sclerosis, with disease features including a long latency and clinical characteristics indistinguishable from idiopathic disease. Cross-sectional 'current' occupational data underestimate cumulative occupational silica exposure. PMID- 10833114 TI - Proximal shift of colorectal cancer in the Australian Capital Territory over 20 years. AB - BACKGROUND: Several studies in other countries have demonstrated a change in subsite distribution of colorectal cancer, with increasing proximal cancers. Confirmation of such a change in Australia would have implications for screening and diagnosis of colorectal cancer. AIMS: To determine whether there has been an increase in the proportion of proximal colorectal cancers in Australia, and whether there have been changes in other clinical and pathological aspects of colorectal cancer. METHODS: A study of the hospital files of patients with colorectal cancer diagnosed and treated at all hospitals in the Australian Capital Territory (ACT) between 1989 and 1995 was compared with data from a published study of patients diagnosed between 1969 and 1976. RESULTS: There was a proximal shift of cancers with a significant increase in the proportion of tumours in the hepatic flexure, ascending colon and caecum, more marked for females than males. There was a corresponding reduction in distal colorectal cancers. Time from onset of symptoms to diagnosis decreased, risk factors for colorectal cancer were noted more frequently, and endoscopy replaced barium enema X-ray as the main diagnostic modality. The resectability of cancers increased, stay in hospital and 30 day mortality declined. Despite apparent earlier presentation and improved surgical resectability, the proportion of patients with localised disease (Dukes' stage A and B) had not changed significantly. CONCLUSIONS: We have detected a number of changes in clinical and pathological aspects of colorectal cancer over a 20 year period in the ACT, including a proximal shift in the subsite distribution of colorectal cancer. These changes suggest that proximal and distal colorectal cancers may have a different aetio pathogenesis, and have implications for the investigation of patients with suspected colorectal cancer and in screening high-risk groups. PMID- 10833115 TI - Improved coronary artery flow after coronary angioplasty in patients with unstable angina. AB - BACKGROUND: Coronary artery flow is impaired after myocardial infarction but there is limited information regarding coronary flow in unstable angina. AIM: To assess baseline coronary artery flow and the effects of coronary angioplasty on coronary flow in patients with unstable angina. METHODS: Twenty-one patients with unstable angina with a culprit lesion suitable for coronary angioplasty were enrolled in the study. Coronary flow was assessed with the Thrombolysis In Myocardial Infarction (TIMI) grade and the Corrected TIMI Frame Count (CTFC) pre and post angioplasty. RESULTS: Baseline flow was impaired in the culprit artery compared to the non culprit artery (42.0+/-28.1 vs 25.3+/-7.0 frames, p<0.02). Pre angioplasty coronary flow was TIMI grade 2 in 52% and TIMI grade 3 in 48% of patients. Post angioplasty flow improved with TIMI grade 2 flow in 5% and TIMI grade 3 in 95%. After angioplasty coronary flow improved from 42.0+/-28.1 frames to 21.6+/-16.3 (p=0.0001). The culprit coronary stenosis decreased from 74+/-9% pre angioplasty to 28+/-12% after intervention (p=0.0001). CONCLUSIONS: Angioplasty and stenting of the culprit vessel restores normal coronary flow in most patients with unstable angina. This suggests that impaired flow in unstable angina is predominantly related to the culprit lesion residual stenosis. PMID- 10833116 TI - Helicobacter pylori eradication with lansoprazole, amoxycillin and clarithromycin: testing an ideal regimen in a multicultural south east Asian population and examining factors potentially influencing eradication. AB - BACKGROUND: From European and North American data, it is recommended in the Asia Pacific consensus statement, that one week therapy with a proton pump inhibitor, amoxycillin and clarithromycin be used for Helicobacter pylori eradication, in areas of high metronidazole resistance. The efficacy of this regimen is unknown in Singapore. AIM: To assess the efficacy, safety and compliance of an H. pylori eradication regimen and examine clinical factors that potentially determine eradication. METHODS: Consecutive outpatients from a multicultural, south east Asian, population with H. pylori infection, with or without peptic ulcer, were treated with lansoprazole 30 mg, amoxycillin 1 gm, clarithromycin 500 mg, twice a day for seven days. Eradication was assessed by either rapid urease, histology or urea breath test. Compliance and side effects were recorded. The eradication rate and effect of ethnicity, age, sex, usage of alcohol, smoking and non-steroidal anti-inflammatory drugs, history of ulcer and endoscopic diagnosis on eradication were examined by univariate and multivariate analysis. RESULTS: Of 113 patients, the eradication rate by intention to treat was 98/113 (87%) (95% confidence interval [CI] 80-93%) and per protocol was 98/106 (92%) (95% CI 87-97%). Using Fisher's exact test, eradication was more successful in Chinese (intention to treat and per protocol respectively p=0.02 and p<0.001) compared to non-Chinese. By logistic regression analysis ethnicity was an independent factor associated with eradication success (p=0.0025). Side effects occurred in five (4.4%), resulting in cessation of treatment. CONCLUSIONS: This one week eradication regimen is safe and effective in south east Asians. Chinese ethnicity may be associated with a higher likelihood of eradication success. PMID- 10833117 TI - Cardiovascular disease towards 2000: activities of the West Australian Heart Research Institute. PMID- 10833118 TI - N-3 polyunsaturated fatty acids and cardiac mortality. PMID- 10833119 TI - The rise in acute medical admissions. PMID- 10833120 TI - Investigating pathology utilisation by junior medical staff in a teaching hospital: a qualitative study. PMID- 10833121 TI - Xenotransplantation into humans. PMID- 10833122 TI - Modern medicine and global communicable diseases: new partnerships for progress. PMID- 10833123 TI - Medical services in China. PMID- 10833124 TI - Inclusion body myositis associated with systemic lupus erythematosus (SLE) PMID- 10833125 TI - Obstetric medicine-a new subspecialty. PMID- 10833126 TI - Autoimmune hypothyroidism associated with interferon beta-1b treatment in two patients with multiple sclerosis. PMID- 10833127 TI - Parathyroid hormone related protein (PTHrP) mediated hypercalcaemia complicating enteropancreatic malignancy in multiple endocrine neoplasia type 1 (MEN 1) PMID- 10833128 TI - Possible Epstein-Barr virus encephalitis in an elderly patient. PMID- 10833129 TI - Systemic lupus erythematosus with vasculitis confined to the coronary arteries. PMID- 10833130 TI - Survey of blood cultures from five community hospitals in south-western Sydney, Australia, 1993-1994. PMID- 10833131 TI - Cepacia syndrome occurring following prolonged colonisation with Burkholderia cepacia. PMID- 10833132 TI - The concealed and the revealed: bioethical issues in Europe at the end of the second millenium. AB - Bioethical debate in Europe is partly a reaction to political endeavors and events. In line with the political goal of a united Europe, a European regulation is being sought for medical research and medical ethics ('Convention on Human Rights and Biomedicine'). A certain degree of ambivalence has come to the fore: whilst it does seem possible to reach a consensus (albeit a merely 'diplomatic' consensus) about complicated bioethical issues at an international level when certain controversial matters are excluded or a certain vagueness maintained, new principles are also required at a national level, for example when the medical profession of one state feels obliged to assume a 'local' stance, such as in the sensitive issue of termination of treatment. The individual contributions to this publication, together with various other current fields of bioethical conflict in Europe--especially Germany--are introduced below against a common background, namely that the original dividing line between the concealed and the revealed has shifted. PMID- 10833133 TI - A philosophical and critical analysis of the European convention of bioethics. AB - The Convention for the Protection of Human Rights and Dignity of the Human Being with Regard to the Application of Biology and Medicine is now one of the most important bioethics texts from the point of view of international policy and law. It is the result of five years of discussions and negotiations between the different instances of the Council of Europe. In this article I analyze several problems. First, there are problems of articulation between the Convention and the joint Explanatory Report. The oriented exegesis of the Explanatory Report raises suspicion about the Convention, which appears as a smooth facade for an instrument actually serving ideological positions many people do not share. Second, there are problems of formulation within the Convention. These are mainly problems with articles that state prohibitions without any distinction, relativization, contextualization or sense of evolution. Finally, there are problems of substance, leading to the conclusion that the Convention is not a good illustration of the human rights philosophical tradition in the name of which it has been proclaimed. This tradition is the one of Enlightenment. And when Kant summarizes the motto of Enlightenment, the injunction is "Sapere Aude!": "Dare to know!" It is difficult to hear such a message through the Convention, and the Explanatory Report includes too many passages and sentences that mean the opposite. PMID- 10833134 TI - Respecting, protecting, persons, humans, and conceptual muddles in the bioethics convention. AB - The Convention on Human Rights and Biomedicine confuses respect for a person's right to self-determination with concern about protecting human beings generally. In a legal document, this mixture of deontological with utilitarian considerations undermines what it should preserve: respect for human dignity as the foundation of modern rights-based democracies. Falling prey to the ambiguity of freedom, the Convention blurs the dividing line between morality and the law. The document should be remedied through distinguishing fundamental rights from social 'rights', persons as entitled to the right to self-determination from born humans as entitled to the right to life and from members of the human species as entitled to the morally responsible care of voting majorities. For the cultivating of the required responsibility, the conditions for an adequate public debate should be secured. PMID- 10833135 TI - Human is what is born of a human: personhood, rationality, and an European convention. AB - In the course of its preparation, the 1997 convention on human rights and biomedicine adopted by the Council of Europe instigated a widespread debate. This article examines one of the core issues: the notion of the human being as depicted in the convention. It is argued that according to the convention, this being may exist in three different legal categories, namely 'human life', 'embryo', and 'personhood', each furnished with an inherent set of somewhat different rights, yet none of them clearly defined, thus leaving it to domestic law to regulate at what point a human being belongs to which category. While this approach is understandable from a political point of view, it creates a vicious circle, since law thereby has to define its own foundation and, in the case of the convention, to protect a being that it cannot define. It appears that this form of life is seen rather as a given entity, taking precedence over the interests of society and science, and its dignity and identity forming criteria for the subsequent systems of culture, simply because this life is human and nothing else. Thus, the convention approaches a natural law position. PMID- 10833136 TI - Palliative care versus euthanasia. The German position: the German General Medical Council's principles for medical care of the terminally ill. AB - In September 1998 the Bundesarztekammer, i.e., the German Medical Association, published new principles concerning terminal medical care. Even before publication, a draft of these principles was very controversial, and prompted intense public debate in the mass media. Despite some of the critics' suspicions that the principles prepared the way for liberalization of active euthanasia, euthanasia is unequivocally rejected in the principles. Physician-assisted suicide is considered to violate professional medical rules. In leaving aside some of the notions customarily used in the euthanasia debate, e.g., passive euthanasia, the principles emphasize the obligation of physicians to offer and the right of patients to receive palliative care. The principles explicitly list modalities of basic treatment that are indispensable in all cases, such as the obligation to satisfy hunger and thirst. This statement is meant to resolve the dispute on nutrition and hydration at the end of life, as it shifts the focus of care from maintaining physiological parameters to satisfying subjective needs. For patients in a persistent vegetative state, artificial feeding is held to be obligatory. Yet, the principles make reference to recent German jurisdiction which permit the stopping of artificial feeding if it is in accordance with the patient's presumed will. Additionally, the wording concerning this issue is found to remain unclear. Patients' autonomy is strengthened by explicitly welcoming advance directives as a means to ascertain patients' wills. The principles mark some changes compared to earlier documents. They deserve careful analysis and should be considered in the international debate on issues concerning the end of life. PMID- 10833137 TI - A right to life for the unborn? The current debate on abortion in Germany and Norbert Hoerster's legal-philosophical justification for the right to life. AB - Rights to life for unborn humans and to abortion with impunity are incompatible. This observation by the German legal philosopher Norbert Hoerster contains a fundamental criticism of the state regulation on abortion in Germany. The regulation regards abortion as unlawful, but declines to prosecute if the abortion is conducted within the first three months of pregnancy and the pregnant woman received counseling at least three days prior to terminating the pregnancy. In contrast to the German legislature, Hoerster is in favor of setting the beginning of a right to life at birth. With this suggestion and the consequent demand for a general legalization of abortion, Hoerster himself has become the target of harsh criticism. The following article analyzes Hoerster's position and that of his opponents against the background of the current abortion debate in Germany. The consequences for dealing with the handicaps of Hoerster's suggested regulations will also be addressed. PMID- 10833138 TI - Patients or customers: ethical limits of market economy in health care. AB - There is a move away from a market economy in health care in the United States and a move towards such a market in Germany. This article tries to make explicit what underlies the moral intuition that there is a tension between a market economy and health care. First, health care is analyzed in terms of the economic theory of the market and incompatibilities are described. The moral problem is identified as the danger of liquefying the distinction between persons and things. The basic moral intuition seems to be the classical social contract: as a functioning market is governed by the principle of commutative justice, free riders have to be kept away, which is achieved by coercion that is not provided by the market; coercion can be justified by a social contract. The special moral problems of a social contract for health care are discussed. It is argued that public coercion in order to collect contributions for essential health care is justified. PMID- 10833139 TI - Principles of the German Medical Association concerning terminal medical care. PMID- 10833140 TI - Convention for the Protection of Human Rights and Dignity of the Human Being with Regard to the Application of Biology and Medicine: Convention on Human Rights and Biomedicine. PMID- 10833141 TI - Distraction osteogenesis for mandibular advancement. AB - The purpose of this study was to investigate the possibilities of distraction osteogenesis to correct mandibular hypoplasia. Fourteen young patients (mean age 14.1 years) with a proven resistance to initial, functional orthodontic therapy, were treated by means of bilateral intraoral distractors. The corticotomy was performed in the region of the third molar. The latency time was six days and the stabilization period six weeks. In all cases the planned lengthening of the mandible and class 1 occlusion were achieved. Seven patients required additional elastic band traction to close a mild open-bite directly after active distraction. In the first seven patients, insufficient mobilization at the site of the corticotomy had resulted in a broken distraction rod in two patients and incomplete distraction of the lingual cortex in one patient. Adequate mobilization at the site of the corticotomy prevented these problems in later cases. No permanent sensory disturbances were seen. Twelve patients finished their orthodontic treatment within six months after distraction. PMID- 10833142 TI - The reproducibility of the head position for a laser scan using a novel morphometric analysis for orthognathic surgery. AB - The aim of this study was to evaluate the reproducibility of the head position for a three-dimensional soft tissue laser scan (lasergraph) using thin-plate splines, for orthognathic surgery planning and follow-up. 60 laser scans of five subjects (12 scans per subject) were obtained at specified intervals. The head was positioned in the lateral view using a spirit level, an engineering device for setting horizontal surfaces, to adjust the Frankfort horizontal plane parallel to the ground. The projection of a narrow beam of longitudinal laser light was used to adjust the axial plane for the frontal view. These scanned images (lasergraphs) were digitised and the co-ordinates of the landmarks recorded. The digitised laser scans were compared using thin-plate splines analysis. The mean difference between the scans due to variations in head position was 0.0135 +/- 0.0109 g x cm2/ sec2 in the lateral view and 0.0090 +/- 0.0054 g x cm2/sec2 in the frontal view. This represents an overall distortion error of less than 2% when following up the surgical change of a typical bimaxillary osteotomy case with 6 mm maxillary advancement and 3 mm mandibular set-back. It is concluded that facial laser scans (lasergraphs) with the Frankfort horizontal plane set using a head rest and spirit level, and the axial plane set using projection of a vertical laser light on the facial midline, are highly reproducible. PMID- 10833143 TI - Pulsatile expansion therapy for orbital enlargement. AB - Experimental and clinical investigations have documented the modulatory role of the globe in the development of the orbit. In cases of absence or early loss of the globe, severe hypoplasia of the orbit and midface has been reported by several authors. Statical conformers and orbital osteotomies have been used to correct the resulting facial asymmetry. When replacing such conformers by increasingly larger ones, orbital structures are negatively influenced by the repeated trauma of surgical interventions. Simulating the modulatory role of the globe on orbital growth was the objective when developing a pulsatile device for orbital enlargement in cases of anophthalmia and microrbitism. The design, application and preliminary experience with a dynamic, pulsatile expanding system are presented. PMID- 10833144 TI - Reconstruction of the contracted ocular socket with free full-thickness mucosa graft. AB - Patients who have undergone enucleation of the eye can be treated with glass eye prostheses, provided retention is adequate. Inadequate retention due to contraction of the conjunctival mucosa is a common problem which frequently affects the lower fornix first. This can be corrected using a free full-thickness buccal mucosa graft. Indications, operative procedures and postoperative results are presented. PMID- 10833145 TI - Peri-implant soft tissue maintenance in patients with craniofacial implant retained prostheses. AB - The aim of this pilot study was to describe the effectiveness of a hygiene protocol prescribed for patients receiving craniofacial implant retained prostheses. Eleven subjects receiving either orbital or auricular prostheses were instructed by a hygienist in debris removal procedures. Patients were re-examined on at least four occasions over the following 18 months, and tissue health around the implant abutments was evaluated using standard criteria. In most cases, adequate debris removal was demonstrated, particularly when hygiene procedures were reinforced at the second follow-up visit. Barriers to maintenance of tissue health included inadequate space between fixtures and thickness of skin around abutments. Occasionally, prostheses had to be replaced due to inappropriate cleaning methods. The intensive hygiene regimen helped maintain tissue health around implant abutments, although it was demanding in terms of professional time. PMID- 10833146 TI - The lateral upper arm free flap for intraoral reconstruction. AB - Twenty-three consecutive patients who were reconstructed with a lateral upper arm free flap (LUFF) were examined especially concerning functional and morphological results at the recipient and donor sites. There were 22 intraoral and one upper oesophageal reconstruction after radical laryngectomy. The LUFF rendered good functional and esthetic results except for one case of complete and one case of incomplete flap necrosis due to vascular insufficiency of the supplying vessel of the neck. There was some sensory deficit of the donor site (n=10), but no radial nerve injury or conspicuous scarring. Recipient site dehiscence occurred in two cases and a temporary orocervical fistula was seen in one case. Oral function was maintained due to the thin and pliable flap. Excellent flap adaptation to the adjacent tissue was obtained in eight cases of complete loss of lingual attached gingiva in the molar region and in four cases of loss of buccal attached gingiva. The success and functional results of LUFF were comparable to the results of 14 cases in which radial forearm free flaps (RFFF) were used. Although the length of the pedicle and the diameter of the vessels in LUFF are smaller than in RFFF, neither pedicle length nor vessel diameter proved to be a problem. Extent of scarring and risk of vascular compromise proved to be less as compared to RFFF. LUFF is, therefore, the flap of choice for intraoral soft tissue reconstruction and it is advised to reserve RFFF for cases in which LUFF fails. PMID- 10833147 TI - Histomorphometric analysis of irradiated recipient vessels and transplant vessels of free flaps in patients undergoing reconstruction after ablative surgery. AB - The aim of the study was to investigate, histomorphometrically, quantitative and qualitative changes in irradiated neck recipient vessels and transplant vessels used for microsurgical anastomoses in free flaps in patients undergoing preoperative radiotherapy and chemotherapy. In 55 patients receiving 42 radial forearm flaps, 6 latissimus dorsi flaps, 6 osteomyocutaneous fibula grafts and 1 lateral arm flap, a total of 220 vessels were obtained from neck recipient vessels and transplant vessels during anastomosis. Three groups were formed: Group 1 (16 patients) treated with no radiotherapy or chemotherapy; Group 2 (20 patients) treated with preoperative irradiation (40-50 Gy) and chemotherapy (800 mg/m2 5-FU and 20 mg/m2 cisplatin) 1.5 months prior to surgery; Group 3 (19 patients) treated with radiotherapy (60-70 Gy) (median interval 78.7 months; IQR 31.3 months) prior to surgery. From each of the 220 vessel specimens, 3 sections each were histomorphometrically investigated, both qualitatively and quantitatively. To evaluate these changes as a function of age, radiation dose and chemotherapy, a statistical analysis was performed using analysis of covariance and chi-square tests. In Group 3, qualitative changes (intima dehiscence, hyalinosis) were found in recipient arteries significantly more frequently (25%, P=0.009) than in Groups 1 and 2. For Group 3 recipient arteries, histomorphometry revealed a significant decrease in the ratio of media area/total vessel area (median 0.53, IQR 0.10) in comparison with Group 1 (P= 0.02) (median 0.60, IQR 0.29) and Group 2 (P=0.046) (median 0.59, IQR 0.10). No significant differences were found between the vessels of Groups 1 and 2 (P= 0.48). Age and chemotherapy did not appear to have a significant influence on vessel changes in this study. PMID- 10833148 TI - Rapid tumor necrosis induced by electrochemotherapy with intratumoral injection of bleomycin in a hamster tongue cancer model. AB - This study evaluated the effects of electrochemotherapy (ECT) with intratumoral injection of bleomycin (BLM) on the chemically induced tongue cancer model in the hamster. Intratumoral injection of BLM followed by high-voltage electrical treatment induced rapid necrosis of the tumor within 48 hours and subsequent rapid tumor volume reduction. Three weeks after the ECT with BLM, 3 of the 6 animals showed no palpable tumor, while no antitumor effects were observed in the control groups. Because of the remarkable antitumor effect with no major observed side effects, we concluded that ECT combined with intratumoral injection of BLM has the potential to enhance treatment results for tongue cancer. PMID- 10833149 TI - Carcinomatous transformation of a sublingual dermoid cyst. A case report. AB - Sublingual dermoid cysts are rare lesions. Typically they present as slow-growing masses that may cause elevation of the tongue, interference with speech or the appearance of a double chin. We report the first case of malignant transformation to squamous cell carcinoma of a long-standing sublingual dermoid cyst. PMID- 10833150 TI - Transoral approach for large pituitary adenoma using Le Fort I osteotomy with mandibulotomy. A case report. AB - A patient is presented with a large pituitary adenoma that was successfully treated with a Le Fort I osteotomy in combination with mandibulotomy. PMID- 10833151 TI - The incidence of cysts and tumors around impacted third molars. AB - The objective of this retrospective analysis was to determine the incidence of the development of cysts and tumors around third molars and to discuss some relevant issues in relation to the removal of asymptomatic, impacted third molars. 9994 impacted third molars, removed in 7582 patients, formed the basis of this study. The analysis revealed 231 cysts (2.31%) and 79 tumors (0.79%), including 7 benign tumors (0.77%) and two malignant tumors (0.02%). The incidence of cysts and tumors around impacted third molars was 3.10%. PMID- 10833152 TI - Rhabdomyoma of the base of the tongue. A case report. AB - A case of rhabdomyoma in the base of the tongue that caused dysphagia and sleep apnea is presented. Diagnosis was accomplished by means of fine needle aspiration biopsy and MRI. The tumor was completely removed by a tongue midline split. PMID- 10833153 TI - Congenital lipomatoid mass of the tongue. AB - A congenital lipomatoid mass of the tongue in a 5-year-old boy is reported. PMID- 10833154 TI - A comparative study on meniscectomy and autogenous graft replacement of the Rhesus monkey temporomandibular joint articular disc--Part I. AB - The aims of this study were to compare the effects of meniscectomy and autogenous graft replacement on the functions and histology of the Rhesus monkey temporomandibular joint. Eight matured Rhesus monkeys (Macaca mulatta) were used for the first part of the study to compare firstly the results of meniscectomy without replacement and secondly, replacement of the disc with a fascia lata graft. The results showed that unilateral meniscectomy in Rhesus monkeys did not result in significant morbidity. Degenerative changes (osteoarthritis) were observed in all the experimental joints with and without replacement of the articular discs. Fascia lata did not survive after grafting and did not reliably induce re-formation of an articular disc-like structure. PMID- 10833155 TI - A comparative study on meniscectomy and autogenous graft replacement of the rhesus monkey temporomandibular joint articular disc--Part II. AB - This study investigated the effects of free auricular cartilage and pedicled temporalis myofascial flap replacement of the temporomandibular joint articular disc in eight rhesus monkeys (Macaca mulatta). The eight animals were divided into two groups (four animals in each group) for replacement of the disc with either auricular cartilage graft or pedicled temporalis myofascial graft. Degenerative changes (osteoarthritis) were observed in all the experimental joints. Despite degeneration of grafted cartilage tissue, histologic and clinical responses after auricular cartilage grafting were favourable. Pedicled temporalis myofascial flap resulted in re-formation of a disc-like interpositional structure in most of the studied joints with the most favourable histologic response. PMID- 10833156 TI - A patient who developed necrosis of the temporal lobe after irradiation of the parotid gland for pleomorphic adenoma. PMID- 10833157 TI - Information economics and the Internet. AB - Information economics offers insights into the dynamics of information across networked systems like the Internet. An information marketplace is different from other marketplaces because an information good is not actually consumed and can be reproduced and distributed at almost no cost. For information producers to remain profitable, they will need to minimize their exposure to competition. For example, information can be sold by charging site access rather than information access fees, or it can be bundled with other information or "versioned." For information consumers, a variation of Malthus' law predicts that the exponential growth in information will mean that specific information will become increasingly expensive to find, because search costs will grow but human attention will remain limited. Furthermore, the low cost of creating poor-quality information on the Web means that the low-quality information may eventually swamp high-quality resources. The use of reputable information portals on the Web, or smart search technologies, may help in the short run, but it is unclear whether an "information famine" is avoidable in the longer term. PMID- 10833158 TI - Biomedicine's electronic publishing paradigm shift: copyright policy and PubMed Central. AB - Biomedical publishing stands at a crossroads. The traditional print, peer reviewed, subscription journal has served science well but is now being called into question. Because of spiraling print journal costs and the worldwide acceptance of the Internet as a valid publication medium, there is a compelling opportunity to re-examine our current paradigm and future options. This report illustrates the conflicts and restrictions inherent in the current publishing model and examines how the single act of permitting authors to retain copyright of their scholarly manuscripts may preserve the quality-control function of the current journal system while allowing PubMed Central, the Internet archiving system recently proposed by the director of the National Institutes of Health, to simplify and liberate access to the world's biomedical literature. PMID- 10833159 TI - Biomedical publishing and the internet: evolution or revolution? AB - The Internet is challenging traditional publishing patterns. In the biomedical domain, medical journals are providing more and more content online, both free and for a fee. Beyond this, however, a number of commentators believe that traditional notions of copyright and intellectual property ownership are no longer suited to the information age and that ownership of copyright to research reports should be and will be wrested from publishers and returned to authors. In this paper, it is argued that, although the Internet will indeed profoundly affect the distribution of biomedical research results, the biomedical publishing industry is too intertwined with the research establishment and too powerful to fall prey to such a copyright revolution. PMID- 10833160 TI - From paper to electron: how an STM journal can survive the disruptive technology of the Internet. AB - The Internet represents a different type of technology for publishers of scientific, technical, and medical journals. It is not a technology that sustains current markets and creates new efficiencies but is, rather, a disruptive technology that could radically alter market forces, profit expectations, and business models. This paper is a translation and amplification of the research done in this area, applied to a large-circulation new science journal, Pediatrics. The findings suggest that the journal of the future will be electronic, have a less volatile cost structure, be supported more by services than by content, be less able to rely on subscription revenues, and abandon certain elements of current value networks. It also provides a possible framework for other publishers to use to evaluate their own journals relative to this disruptive technology. PMID- 10833161 TI - Solutions to challenges facing a university digital library and press. AB - During the creation of a university digital library and press intended to serve as a medical reference and education tool for health care providers and their patients, six distinct and complex digital publishing challenges were encountered. Over nine years, through a multidisciplinary approach, solutions were devised to the challenges of digital content ownership, management, mirroring, translation, interactions with users, and archiving. The result is a unique, author-owned, internationally mirrored, university digital library and press that serves as an authoritative medical reference and education tool for users around the world. The purpose of this paper is to share the valuable digital publishing lessons learned and outline the challenges facing university digital libraries and presses. PMID- 10833162 TI - Bayesian communication: a clinically significant paradigm for electronic publication. AB - OBJECTIVE: To develop a model for Bayesian communication to enable readers to make reported data more relevant by including their prior knowledge and values. BACKGROUND: To change their practice, clinicians need good evidence, yet they also need to make new technology applicable to their local knowledge and circumstances. Availability of the Web has the potential for greatly affecting the scientific communication process between research and clinician. Going beyond format changes and hyperlinking, Bayesian communication enables readers to make reported data more relevant by including their prior knowledge and values. This paper addresses the needs and implications for Bayesian communication. FORMULATION: Literature review and development of specifications from readers', authors', publishers', and computers' perspectives consistent with formal requirements for Bayesian reasoning. RESULTS: Seventeen specifications were developed, which included eight for readers (express prior knowledge, view effect size and variability, express threshold, make inferences, view explanation, evaluate study and statistical quality, synthesize multiple studies, and view prior beliefs of the community), three for authors (protect the author's investment, publish enough information, make authoring easy), three for publishers (limit liability, scale up, and establish a business model), and two for computers (incorporate into reading process, use familiar interface metaphors). A sample client-only prototype is available at http://omie.med.jhmi.edu/bayes. CONCLUSION: Bayesian communication has formal justification consistent with the needs of readers and can best be implemented in an online environment. Much research must be done to establish whether the formalism and the reality of readers' needs can meet. PMID- 10833163 TI - GeneClinics: a hybrid text/data electronic publishing model using XML applied to clinical genetic testing. AB - GeneClinics is an online genetic information resource consisting of descriptions of specific inherited disorders ("disease profiles") as well as information on the role of genetic testing in the diagnosis, management, and genetic counseling of patients with these inherited conditions. GeneClinics is intended to promote the use of genetic services in medical care and personal decision making by providing health care practitioners and patients with information on genetic testing for specific inherited disorders. GeneClinics is implemented as an object oriented database containing a combination of data and semistructured text that is rendered as HTML for publishing a given "disease profile" on the Web. Content is acquired from authors via templates, converted to an XML document reflecting the underlying database schema (with tagging of embedded data), and then loaded into the database and subjected to peer review. The initial implementation of a production system and the first phase of population of the GeneClinics database content are complete. Further expansion of the content to cover more disease, significant scaling up of rate of content creation, and evaluation redesign are under way. The ultimate goal is to have an entry in GeneClinics for each entry in the GeneTests directory of medical genetics laboratories-that is, for each disease for which clinical genetic testing is available. PMID- 10833164 TI - When conversation is better than computation. AB - While largely ignored in informatics thinking, the clinical communication space accounts for the major part of the information flow in health care. Growing evidence indicates that errors in communication give rise to substantial clinical morbidity and mortality. This paper explores the implications of acknowledging the primacy of the communication space in informatics and explores some solutions to communication difficulties. It also examines whether understanding the dynamics of communication between human beings can also improve the way we design information systems in health care. Using the concept of common ground in conversation, proposals are suggested for modeling the common ground between a system and human users. Such models provide insights into when communication or computational systems are better suited to solving information problems. PMID- 10833165 TI - Representing knowledge: introduction to the Cornerstone I session at the 1999 AMIA Annual Symposium PMID- 10833166 TI - From data to knowledge through concept-oriented terminologies: experience with the Medical Entities Dictionary. AB - Knowledge representation involves enumeration of conceptual symbols and arrangement of these symbols into some meaningful structure. Medical knowledge representation has traditionally focused more on the structure than the symbols. Several significant efforts are under way, at local, national, and international levels, to address the representation of the symbols though the creation of high quality terminologies that are themselves knowledge based. This paper reviews these efforts, including the Medical Entities Dictionary (MED) in use at Columbia University and the New York Presbyterian Hospital. A decade's experience with the MED is summarized to serve as a proof-of-concept that knowledge-based terminologies can support the use of coded patient data for a variety of knowledge-based activities, including the improved understanding of patient data, the access of information sources relevant to specific patient care problems, the application of expert systems directly to the care of patients, and the discovery of new medical knowledge. The terminological knowledge in the MED has also been used successfully to support clinical application development and maintenance, including that of the MED itself. On the basis of this experience, current efforts to create standard knowledge-based terminologies appear to be justified. PMID- 10833167 TI - Clinical classification and terminology: some history and current observations. AB - The evolution of health terminology has undergone glacial transition over time, although this pace has quickened recently. After a long history of near neglect, unimaginative structure, and factitious development, health terminologies are in an era of unprecedented importance, sophistication, and collaboration. The major highlights of this history are reviewed, together with important intellectual advances in health terminology development. The inescapable conclusion is that we are amidst a major revolution in the role and capabilities of health terminologies, entering an age of large-scale systems for health concept representation with international implications. PMID- 10833168 TI - Are medical informatics and nursing informatics distinct disciplines? The 1999 ACMI debate. AB - The 1999 debate of the American College of Medical Informatics focused on the proposition that medical informatics and nursing informatics are distinctive disciplines that require their own core curricula, training programs, and professional identities. Proponents of this position emphasized that informatics training, technology applications, and professional identities are closely tied to the activities of the health professionals they serve and that, as nursing and medicine differ, so do the corresponding efforts in information science and technology. Opponents of the proposition asserted that informatics is built on a re-usable and widely applicable set of methods that are common to all health science disciplines, and that "medical informatics" continues to be a useful name for a composite core discipline that should be studied by all students, regardless of their health profession orientation. PMID- 10833169 TI - Design and implementation of a national clinical trials registry. AB - The authors have developed a Web-based system that provides summary information about clinical trials being conducted throughout the United States. The first version of the system, publicly available in February 2000, contains more than 4,000 records representing primarily trials sponsored by the National Institutes of Health. The impetus for this system has come from the Food and Drug Administration (FDA) Modernization Act of 1997, which mandated a registry of both federally and privately funded clinical trials "of experimental treatments for serious or life-threatening diseases or conditions." The system design and implementation have been guided by several principles. First, all stages of system development were guided by the needs of the primary intended audience, patients and other members of the public. Second, broad agreement on a common set of data elements was obtained. Third, the system was designed in a modular and extensible way, and search methods that take extensive advantage of the National Library of Medicine's Unified Medical Language System (UMLS) were developed. Finally, since this will be a long-term effort involving many individuals and organizations, the project is being implemented in several phases. PMID- 10833170 TI - Electronic publishing of scholarly communication in the biomedical sciences. PMID- 10833171 TI - Presentation of the Morris F. Collen Award to Joshua Lederberg, PhD. PMID- 10833172 TI - What is Sure Start? PMID- 10833173 TI - Recent advances in management of acute leukaemia. AB - There have been significant improvements in the outlook for children with acute leukaemia but these advances are only available to a minority of the world's children. There is still room for improvements in conventional chemotherapy and these need evaluation in randomised trials. The role of bone marrow transplants in first remission is evolving as chemotherapy becomes more effective. New treatments are needed for relapsed patients. Molecular diagnosis has refined the assessment of prognosis but the extra value afforded by measurement of minimal residual disease is not clear. International collaboration is needed to evaluate treatment for rare subtypes of leukaemia. PMID- 10833174 TI - Does home visiting improve parenting and the quality of the home environment? A systematic review and meta analysis. AB - AIMS: To evaluate the effectiveness of home visiting programmes on parenting and quality of the home environment. DESIGN: Systematic review of the literature of randomised controlled trials and quasi-experimental studies evaluating home visiting programmes involving at least one postnatal visit. SUBJECTS: Thirty four studies reported relevant outcomes; 26 used participants considered to be at risk of adverse maternal or child health outcomes; two used preterm or low birth weight infants; and two used infants with failure to thrive. Only eight used participants not considered to be at risk of adverse child health outcomes. RESULTS: Seventeen studies reported Home Observation for Measurement of the Environment (HOME) scores, 27 reported other measures of parenting, and 10 reported both types of outcome. Twelve studies were entered into the meta analysis. This showed a significant effect of home visiting on HOME score. Similar results were found after restricting the analyses to randomised controlled trials and to higher quality studies. Twenty one of the 27 studies reporting other measures of parenting found significant treatment effects favouring the home visited group on a range of measures. CONCLUSIONS: Home visiting programmes were associated with an improvement in the quality of the home environment. Few studies used UK health visitors, so caution must be exercised in extrapolating the results to current UK health visiting practice. Further work is needed to evaluate whether UK health visitors can achieve similar results. Comparisons with similar programmes delivered by paraprofessionals or community mothers are also needed. PMID- 10833175 TI - Incidence and cause of fractures in European districts. AB - AIMS: To compare fracture rates in European districts. SETTING: Geographically defined areas of Wales (Swansea and Neath Port Talbot), Norway (Harstad, Trondheim, Stavanger, and Drammen), Sweden (Jamtlands), and Finland (Porvoo). METHODS: Surveillance of fractures at emergency departments and hospitals and linkage with population data. Comparison of age adjusted and crude rates. Calculation of confidence intervals for ratios. RESULTS: A total of 4113 fractures occurred in 167 560 children during 1996. Fracture rates in south Wales (36 per 1000) were substantially higher than in Scandinavian districts (which were similar). Limiting analysis to the most severe injuries to correct for the possibility of ascertainment bias reduced some of the excess rate in Wales: the Welsh:Scandinavian fracture ratio was 1.82 (95% confidence interval: 1.64 to 2.03). CONCLUSIONS: Fracture rates in Welsh children are substantially higher than in Scandinavian children. PMID- 10833176 TI - Sudden unexpected non-violent death between 1 and 19 years in north Spain. AB - AIMS: To study the epidemiological, clinical, and pathological characteristics of sudden unexpected non-violent deaths between 1 and 19 years. METHODS: Population based observational study of all sudden unexpected non-violent deaths between 1 and 19 years, from 1990 to 1997, in a north Spain county (Bizkaia). In each case, clinical information and circumstances of death were obtained and a complete forensic autopsy was performed. RESULTS: There were 34 sudden unexpected non violent deaths. The mortality rate was 1.7 per 100 000 persons per year (representing 9% of the mortality rate of all non-violent deaths). In 10 cases the cause of death was cardiac, in 13 cases extracardiac, and 11 cases were unexplained. In 17 cases there were no pathological antecedents and in nine cases death occurred in patients with known disorders. Syncopes prior to death were present in five cases. Exercise related deaths occurred in seven cases (mainly associated with cardiac causes). Eight cases were "death in bed". PMID- 10833177 TI - Weight gain and sudden infant death syndrome: changes in weight z scores may identify infants at increased risk. AB - AIMS: To investigate patterns of infant growth that may influence the risk of sudden infant death syndrome (SIDS). DESIGN: Three year population based case control study with parental interviews for each death and four age matched controls. Growth was measured from prospective weight observations using the British 1990 Growth Reference. SETTING: Five regions in England (population greater than 17 million, more than 470 000 live births over three years). SUBJECTS: 247 SIDS cases and 1110 controls. RESULTS: The growth rate from birth to the final weight observation was significantly poorer among the SIDS infants despite controlling for potential confounders (SIDS mean change in weight z score (deltazw) = -0.38 (SD 1.40) v controls = +0.22 (SD 1.10), multivariate: p < 0.0001). Weight gain was poorer among SIDS infants with a normal birth weight (above the 16th centile: odds ratio (OR) = 1.75, 95% confidence interval (CI) 1. 48-2.07, p < 0.0001) than for those with lower birth weight (OR = 1. 09, 95% CI 0.61-1.95, p = 0.76). There was no evidence of increased growth retardation before death. CONCLUSIONS: Poor postnatal weight gain was independently associated with an increased risk of SIDS and could be identified at the routine six week assessment. PMID- 10833178 TI - Current approaches to the management of primary hyperoxaluria. PMID- 10833179 TI - Europe calling PMID- 10833180 TI - Psychogenic non-epileptic seizures: management and prognosis. AB - AIM: To determine the outcome and identify predictive factors in children with psychogenic non-epileptic seizures (PNES). METHOD: The biographies of 35 children with PNES, attending a tertiary paediatric neurology centre, were reviewed. RESULTS: Thirty five children attending the department between 1987 and 1997 were evaluated at a mean follow up of 4.6 years. The age range was 6-18 years. Twenty four were girls and 11 were boys. Eleven patients had a diagnosis of epilepsy with PNES, the remainder having PNES alone. Cause fell into four categories: a history of violence, abuse, or neglect; a high level of anxiety; dysfunctional family relationships; and attention seeking or avoidance behaviour. Management in all but five cases involved assessment and follow up by a child psychologist or child psychiatrist. The outcome was encouraging, with 66% of patients becoming PNES free. A further 23% have > 50% reduction in the frequency of PNES. Only two have had no reduction. Outcome was best in the group without epilepsy. CONCLUSION: These results suggest that the prognosis of PNES is better in children than in adults, perhaps because causes are more likely to be external to the child, more easily identified, and more amenable to prompt intervention. The importance of good assessment, good communication, and a treatment plan that includes both symptom management and addressing the precipitating and perpetuating factors is emphasised. PMID- 10833181 TI - Hepatocellular carcinoma in glycogen storage disease type IV. AB - A 13 year old patient with juvenile type IV glycogen storage disease died of the complications of hepatocellular carcinoma. To our knowledge this is the first reported case of hepatocellular carcinoma in association with type IV glycogen storage disease. PMID- 10833182 TI - Protracted neonatal hypertrypsinogenaemia, normal sweat chloride, and cystic fibrosis. AB - The cystic fibrosis (CF) clinical spectrum has greatly expanded in the past few years, including atypical forms with low sweat chloride concentrations. Two cases are presented which suggest that children detected by neonatal CF screening whose trypsinogen concentrations are still raised by the second month of age could, despite a negative sweat test, be affected by an atypical CF with fully expressed pulmonary involvement. PMID- 10833183 TI - Long term follow up of topical mustine treatment for cutaneous langerhans cell histiocytosis. AB - BACKGROUND AND OBJECTIVES: Skin lesions in Langerhans cell histiocytosis (LCH) are often painful and difficult to treat. Topical application of nitrogen mustard (0.02% mechlorethamine hydrochloride, mustine), an alkylating cytostatic agent, has been shown to be effective. There is, however, concern about potentially harmful long term side effects. STUDY DESIGN: In a retrospective study 20 children with LCH (average extent of initial skin involvement: 16.4% body surface) were followed up for an average of 8.3 years after completion of topical mustine therapy. They had received a total of 34 courses (mean duration 14.2 weeks) of topical mustine. Disease status on follow up was assessed according to the Histiocyte Society classification. RESULTS: After mustine was introduced, 16 patients were able to discontinue systemic steroids and/or chemotherapy. Topical mustine was well tolerated in 18 patients, but two developed irritant dermatitis. On follow up, the disease was inactive in 10 patients. Among the children with active disease, six had mild skin disease and four had progressive disease, two of them with skin lesions unresponsive to mustine treatment. Scars confined to areas of formerly active skin disease were found in six patients. There was no evidence of premalignant or malignant skin disease in the treated areas. CONCLUSION: Topical mustine is an effective and safe treatment for skin disease in most children with LCH. Residual scarring was probably a result of the disease itself rather than to mustine. Although no evidence of skin cancer was found in this study, continued long term follow up is advisable. PMID- 10833184 TI - Food entrapped in papilla of Vater: uncommon cause of vomiting. AB - CASE REPORT: A 20 month old girl was admitted for intractable vomiting over several days, with no other symptoms. Family and personal history were not contributive. Clinical and neurological examination, and routine blood tests and investigations (plain abdominal x ray, upper gastrointestinal tract contrast study, abdominal ultrasonography) were normal. The upper gastrointestinal endoscopy showed a mild antral gastritis and the second portion of duodenum was occupied by a tough, fibrous mass partially embedded into the papilla of Vater. The foreign body was removed and proved to be vegetable fibre (pineapple). Symptoms subsided immediately and the child was discharged with gastroprotective therapy. After two months, on endoscopic examination, the signs of gastropathy had cleared; the papilla of Vater was undamaged, but unchomped food debris was again found in the duodenum. DISCUSSION: There are sporadic reports of foreign bodies retained into the papilla of Vater, all of them in adults. This child, though her papilla was tiny, had no jaundice or pancreatitis, unlike most of the reported cases. This is the first report of this finding in a child. The cause of the vomiting was not shown on abdominal ultrasonography or contrast study. It should be added to the list of unusual causes of vomiting. PMID- 10833185 TI - Unicef PMID- 10833186 TI - Diagnosis of Smith-Lemli-Opitz syndrome from stored filter paper blood specimens. AB - BACKGROUND: Smith-Lemli-Opitz (SLO) syndrome is a recessively inheritable metabolic disease with deficiency of cholesterol and accumulation of dehydrocholesterols, caused by a defect in the last step of cholesterol biosynthesis. Biochemical methods for identification of affected individuals, even prenatally, have been developed. Reliable genetic counselling is now possible. AIM: To find a method of proving or disproving whether a child in whom SLO syndrome had been suspected but not confirmed during lifetime had in fact died of the SLO syndrome. METHODS: Lipid extracts of stored filter paper blood specimens collected at the national neonatal metabolic screening were used. The ratio of dehydrocholesterols to cholesterol was measured by combined gas chromatography-mass spectrometry. RESULTS: The ratio of 8-dehydrocholesterol to cholesterol in stored filter paper specimens clearly distinguished affected infants from normal infants. SLO syndrome was thus proven in two children who had died more than seven years earlier. CONCLUSION: It is possible to diagnose SLO syndrome from dried paper specimens, even when the samples were collected more than a decade ago. Genetic counselling is available for families of affected children who died before the discovery of the defect in cholesterol synthesis. PMID- 10833187 TI - Breast milk docosahexaenoic acid (DHA) correlates with DHA status of malnourished infants. AB - AIM: To investigate whether low docosahexaenoic acid (22:6omega3; DHA) status of malnourished, mostly breast fed infants is a result of low omega3 fatty acid intake via breast milk. METHODS: Fatty acid composition of breast milk of eight Pakistani mothers, and of the erythrocytes of their malnourished children was analysed. RESULTS: The milk of the Pakistani mothers contained low percentages of all omega3 and most omega6 fatty acids, compared with milk of Dutch mothers. Breast milk DHA was positively correlated with infant erythrocyte DHA and arachidonic acid (20:4omega6). CONCLUSION: DHA status of these malnourished children is strongly dependent on the omega3 fatty acid intake from breast milk. Augmentation of the infants' omega3 long chain polyunsaturated fatty acid status, or the omega3 and omega6 fatty acid status in general, by supplementation is indicated in deprived circumstances where access to fresh fish is difficult. However, in terms of prevention, maternal supplementation of these long chain polyunsaturated fatty acids, preferably from early pregnancy onwards, may be a better option. PMID- 10833188 TI - Lung deposition of aerosol--a comparison of different spacers. AB - AIMS: To investigate (1) aerosol lung deposition obtained from two small volume conventional spacers (Babyhaler and Aerochamber) and a home made spacer (modified 500 ml plastic cold drink bottle); (2) the effect of using a face mask or mouthpiece; and (3) the relation between age and pulmonary deposition. METHODS: Lung deposition of aerosolised technetium-99m DTPA inhaled via spacer was measured in 40 children aged 3-7 years with stable asthma. Each patient performed sequential randomly assigned inhalations using two spacers. Three studies were performed: Babyhaler compared to Aerochamber (with facemasks); Babyhaler with facemask compared to Babyhaler with mouthpiece; and Babyhaler with mouthpiece compared to a 500 ml bottle. RESULTS: Median lung aerosol deposition from a Babyhaler and Aerochamber with masks were similar (25% v 21%, p = 0.9). Aerosol lung deposition from a Babyhaler with mask compared to a Babyhaler with mouthpiece was equivalent (26% v 26%, p = 0.5). Lung deposition was higher from a 500 ml bottle compared to a Babyhaler in both young (25% v 12.5%, p = 0.005) and older children (42% v 22.5%, p = 0.003). A notable reduction in pulmonary deposition occurred at 50 months of age. CONCLUSION: A Babyhaler or Aerochamber produce equivalent lung deposition of aerosol. There is no difference in lung deposition when a mask or mouthpiece is used. A modified 500 ml plastic bottle produces greater pulmonary aerosol deposition than a conventional small volume spacer. PMID- 10833189 TI - Does adjustment of GFR to extracellular fluid volume improve the clinical utility of cystatin C? AB - BACKGROUND: Cystatin C measurement has been proposed as a replacement for creatinine as a serum measure of glomerular filtration rate (GFR). It has also been suggested that GFR itself should be adjusted to the extracellular fluid volume (ECV) of a child rather than the body surface area (BSA). AIMS: To assess the potential of cystatin C compared to serum creatinine in assessing GFR and to establish whether adjustment of GFR to ECV rather than BSA affects the potential usefulness of cystatin C. METHODS: Cystatin C and plasma creatinine were measured in 64 paediatric patients undergoing 77 (51)Cr-EDTA GFR measurements over a six month period. RESULTS: 1/cystatin C concentrations were more closely related to GFR (median 98 ml/min/1.73 m(2), range 8-172) after adjustment for patient BSA (r = 0.81 versus r = 0.44). 1/Creatinine concentrations appeared to be an inferior estimate of BSA adjusted GFR (r = 0.41), even following the use of the Schwartz formula (r = 0.37). Bland Altman statistics showed cystatin C could still only predict 95% of GFR values to within +/-41 ml/min/1.73 m(2) of the (51)Cr-EDTA method. The relation between GFR and 1/cystatin C was not improved by adjusting (51)Cr-EDTA GFR to ECV rather than BSA (r = 0.76 versus r = 0.81). CONCLUSIONS: Cystatin C appears superior to serum creatinine in paediatric subjects although its performance is unlikely to supplant (51)Cr-EDTA GFR measurement. This performance is not being underestimated because of adjusting GFR to BSA rather than ECV. PMID- 10833190 TI - Parsimony, likelihood, and the role of models in molecular phylogenetics. AB - Methods such as maximum parsimony (MP) are frequently criticized as being statistically unsound and not being based on any "model." On the other hand, advocates of MP claim that maximum likelihood (ML) has some fundamental problems. Here, we explore the connection between the different versions of MP and ML methods, particularly in light of recent theoretical results. We describe links between the two methods--for example, we describe how MP can be regarded as an ML method when there is no common mechanism between sites (such as might occur with morphological data and certain forms of molecular data). In the process, we clarify certain historical points of disagreement between proponents of the two methodologies, including a discussion of several forms of the ML optimality criterion. We also describe some additional results that shed light on how much needs to be assumed about underlying models of sequence evolution in order to successfully reconstruct evolutionary trees. PMID- 10833191 TI - Microsatellite diversity correlated with ecological-edaphic and genetic factors in three microsites of wild emmer wheat in North Israel. AB - This study was conducted to test the effects of internal (genetic) and external factors on allelic diversity at 27 dinucleotide microsatellite (simple sequence repeat [SSR]) loci in three Israeli natural populations of Triticum dicoccoides from Ammiad, Tabigha, and Yehudiyya, north of the Sea of Galilee. The results demonstrated that SSR diversity is correlated with the interaction of ecological and genetic factors. Genetic factors, including genome (A vs. B), chromosome, motif, and locus, affected average repeat number (ARN), variance in repeat number (sigma), and number of alleles (NA) of SSRs, but the significance of some factors varied among populations. Genome effect on SSR variation may result from different motif types, particularly compound (or imperfect) versus perfect motifs, which may be related to different evolutionary histories of genomes A and B. Ecological factors significantly affected SSR variation. Soil-unique and soil specific alleles were found in two edaphic groups dwelling on terra rossa and basalt soils across macro- and microgeographical scales. The largest contributions of genetic and ecological effects were found for diversity of ARN and NA, respectively. Multiple regression indicated that replication slippage and unequal crossing over could be important mutational mechanisms, but their significance varied among motifs. Edaphic stresses may affect the probability of replication errors and recombination intermediates and thus control diversity level and divergence of SSRs. The results may indicate that SSR diversity is adaptive, channeled by natural selection and influenced by both internal and external factors and their interactions. PMID- 10833192 TI - The complete sequence of the mitochondrial genome of the crustacean Penaeus monodon: are malacostracan crustaceans more closely related to insects than to branchiopods? AB - The complete sequence of the mitochondrial genome of the giant tiger prawn, Penaeus monodon (Arthropoda, Crustacea, Malacostraca), is presented. The gene content and gene order are identical to those observed in Drosophila yakuba. The overall AT composition is lower than that observed in the known insect mitochondrial genomes, but higher than that observed in the other two crustaceans for which complete mitochondrial sequence is available. Analysis of the effect of nucleotide bias on codon composition across the Arthropoda reveals a trend with the crustaceans represented showing the lowest proportion of AT-rich codons in mitochondrial protein genes. Phylogenetic analysis among arthropods using concatenated protein-coding sequences provides further support for the possibility that Crustacea are paraphyletic. Furthermore, in contrast to data from the nuclear gene EF1alpha, the first complete sequence of a malacostracan mitochondrial genome supports the possibility that Malacostraca are more closely related to Insecta than to Branchiopoda. PMID- 10833193 TI - Likelihood analysis of phylogenetic networks using directed graphical models. AB - A method for computing the likelihood of a set of sequences assuming a phylogenetic network as an evolutionary hypothesis is presented. The approach applies directed graphical models to sequence evolution on networks and is a natural generalization of earlier work by Felsenstein on evolutionary trees, including it as a special case. The likelihood computation involves several steps. First, the phylogenetic network is rooted to form a directed acyclic graph (DAG). Then, applying standard models for nucleotide/amino acid substitution, the DAG is converted into a Bayesian network from which the joint probability distribution involving all nodes of the network can be directly read. The joint probability is explicitly dependent on branch lengths and on recombination parameters (prior probability of a parent sequence). The likelihood of the data assuming no knowledge of hidden nodes is obtained by marginalization, i.e., by summing over all combinations of unknown states. As the number of terms increases exponentially with the number of hidden nodes, a Markov chain Monte Carlo procedure (Gibbs sampling) is used to accurately approximate the likelihood by summing over the most important states only. Investigating a human T-cell lymphotropic virus (HTLV) data set and optimizing both branch lengths and recombination parameters, we find that the likelihood of a corresponding phylogenetic network outperforms a set of competing evolutionary trees. In general, except for the case of a tree, the likelihood of a network will be dependent on the choice of the root, even if a reversible model of substitution is applied. Thus, the method also provides a way in which to root a phylogenetic network by choosing a node that produces a most likely network. PMID- 10833194 TI - Evolution of the eukaryotic translation termination system: origins of release factors. AB - Accurate translation termination is essential for cell viability. In eukaryotes, this process is strictly maintained by two proteins, eukaryotic release factor 1 (eRF1), which recognizes all stop codons and hydrolyzes peptidyl-tRNA, and eukaryotic release factor 3 (eRF3), which is an elongation factor 1alpha (EF 1alpha) homolog stimulating eRF1 activity. To retrace the evolution of this core system, we cloned and sequenced the eRF3 genes from Trichomonas vaginalis (Parabasalia) and Giardia lamblia (Diplomonada), which are generally thought to be "early-diverging eukaryotes," as well as those from two ciliates (Oxytricha trifallax and Euplotes aediculatus). We also determined the sequence of the eRF1 gene for G. lamblia. Surprisingly, the G. lamblia eRF3 appears to have only one domain, corresponding to EF-1alpha, while other eRF3s (including the T. vaginalis protein) have an additional N-terminal domain, of 66-411 amino acids. Considering this novel eRF3 structure and our extensive phylogenetic analyses, we suggest that (1) the current translation termination system in eukaryotes evolved from the archaea-like version, (2) eRF3 was introduced into the system prior to the divergence of extant eukaryotes, including G. lamblia, and (3) G. lamblia might be the first eukaryotic branch among the organisms considered. PMID- 10833195 TI - A fast algorithm for joint reconstruction of ancestral amino acid sequences. AB - A dynamic programming algorithm is developed for maximum-likelihood reconstruction of the set of all ancestral amino acid sequences in a phylogenetic tree. To date, exhaustive algorithms that find the most likely set of ancestral states (joint reconstruction) have running times that scale exponentially with the number of sequences and are thus limited to very few taxa. The time requirement of our new algorithm scales linearly with the number of sequences and is therefore applicable to practically any number of taxa. A detailed description of the new algorithm and an example of its application to cytochrome b sequences are provided. PMID- 10833196 TI - Phylogenetic analysis under reticulate evolution. AB - The usual assumption that species have evolved from a common ancestor by a simple branching process--where each branch is genetically isolated--has been challenged by the observation of frequent hybridization between species in natural populations. In fact, most plant species are thought to have hybrid origins. This reticulate pattern of species evolution has posed problems in the definition of speciation and in phylogenetic reconstruction, especially when molecular data are used. As a result, hybridization has been largely treated as an evolutionary accident or statistical error in phylogenetic analysis. In this paper, I explicitly incorporate hybridization as an evolutionary occurrence and then conduct phylogenetic reconstruction. I first examine the reticulate evolution under a pure drift model, and then extend the theory to fit a mutation model. A least-squares method is developed for reconstructing a reticulate phylogeny using gene frequency data. The efficacy of the method under the pure drift model is verified via Monte Carlo simulations. PMID- 10833197 TI - Evolution of the Gypsy endogenous retrovirus in the Drosophila melanogaster subgroup. AB - We conducted a phylogenetic survey of the endogenous retrovirus Gypsy in the eight species of the Drosophila melanogaster subgroup. A 362-bp fragment from the integrase gene (int) was amplified, cloned, and sequenced. Phylogenetic relationships of the elements isolated from independent clones were compared with the host phylogeny. Our results indicate that two main lineages of Gypsy exist in the melanogaster subgroup and that vertical and horizontal transmission have played a crucial role in the evolution of this insect endogenous retrovirus. PMID- 10833198 TI - L1 (LINE-1) retrotransposon evolution and amplification in recent human history. AB - L1 (LINE-1) elements constitute a large family of mammalian retrotransposons that have been replicating and evolving in mammals for more than 100 Myr and now compose 20% or more of the DNA of some mammals. Here, we investigated the evolutionary dynamics of the active human Ta L1 family and found that it arose approximately 4 MYA and subsequently differentiated into two major subfamilies, Ta-0 and Ta-1, each of which contain additional subsets. Ta-1, which has not heretofore been described, is younger than Ta-0 and now accounts for at least 50% of the Ta family. Although Ta-0 contains some active elements, the Ta-1 subfamily has replaced it as the replicatively dominant subfamily in humans; 69% of the loci that contain Ta-1 inserts are polymorphic for the presence or absence of the insert in human populations, as compared with 29% of the loci that contain Ta-0 inserts. This value is 90% for loci that contain Ta-1d inserts, which are the youngest subset of Ta-1 and now account for about two thirds of the Ta-1 subfamily. The successive emergence and amplification of distinct Ta L1 subfamilies shows that L1 evolution has been as active in recent human history as it has been found to be for rodent L1 families. In addition, Ta-1 elements have been accumulating in humans at about the same rate per generation as recently evolved active rodent L1 subfamilies. PMID- 10833199 TI - How important is DNA replication for mutagenesis? AB - Rates of mutation and substitution in mammals are generally greater in the germ lines of males. This is usually explained as resulting from the larger number of germ cell divisions during spermatogenesis compared with oogenesis, with the assumption made that mutations occur primarily during DNA replication. However, the rate of cell division is not the only difference between male and female germ lines, and mechanisms are known that can give rise to mutations independently of DNA replication. We investigate the possibility that there are other causes of male-biased mutation. First, we show that patterns of variation at approximately 5,200 short tandem repeat (STR) loci indicate a higher mutation rate in males. We estimate a ratio of male-to-female mutation rates of approximately 1.9. This is significantly greater than 1 and supports a greater rate of mutation in males, affecting the evolution of these loci. Second, we show that there are chromosome specific patterns of nucleotide and dinucleotide composition in mammals that have been shaped by mutation at CpG dinucleotides. Comparable patterns occur in birds. In mammals, male germ lines are more methylated than female germ lines, and these patterns indicate that differential methylation has played a role in male-biased vertebrate evolution. However, estimates of male mutation bias obtained from both classes of mutation are substantially lower than estimates of cell division bias from anatomical data. This discrepancy, along with published data indicating slipped-strand mispairing arising at STR loci in nonreplicating DNA, suggests that a substantial percentage of mutation may occur in nonreplicating DNA. PMID- 10833200 TI - A re-evaluation of 12S ribosomal RNA variability in Drosophila pseudoobscura. AB - Two recent studies have presented conflicting views on variation present within the 294 base third domain of the 12S rRNA gene in the genus Drosophila, and in D. pseudoobscura in particular. One study suggested that this gene is highly invariant across the genus, while another recovered 22 distinct haplotypes from 22 strains of D. pseudoobscura. We have sequenced this gene in numerous lines of D. pseudoobscura and its relatives, noting only two haplotypes in the third domain, and we failed to confirm any of the published sequences. Second, we note that the published sequence divergence between strains of D. pseudoobscura was as great as that documented between distantly related Drosophila species. Third, we show that the published polymorphisms of this region within D. pseudoobscura would disrupt the secondary structure of the resulting molecule. We conclude that the published 12S rRNA sequences of D. pseudoobscura do not accurately reflect variability of the functional gene, and that this gene is relatively invariant in D. pseudoobscura and D. persimilis. PMID- 10833201 TI - Big-benefit mutations in a bacteriophage inhibited with heat. AB - High temperature inhibits the growth of the wild-type bacteriophage phiX174. Three different point mutations were identified that each recovered growth at high temperature. Two affected the major capsid protein (residues F188 and F242), and one affected the internal scaffolding protein (B114). One of the major capsid mutations (F242) is located in a beta strand that contacts B114 in the procapsid during viral maturation, whereas the other capsid mutation (F188) is involved in subunit interactions at the threefold axis of symmetry. Selective coefficients of these mutations ranged from 13.9 to 3.8 in the inhibitory, hot environment, but all mutations reduced fitness at normal temperature. The selective effect of one of the mutations (F242) was evaluated at high temperature in four different genetic backgrounds and exhibited epistasis of diminishing returns: as log fitness of the background genotype increased from -0.1 to 4.1, the fitness boost provided by the F242 mutation decreased from 3.9 to 0. 8. These results support a model in which viral fitness is bounded by an upper limit and the benefit of a mutation is scaled according to the remaining opportunity for fitness improvement in the genome. PMID- 10833202 TI - Why mitochondrial genes are most often found in nuclei. AB - A very small fraction of the proteins required for the propagation and function of mitochondria are coded by their genomes, while nuclear genes code the vast majority. We studied the migration of genes between the two genomes when transfer mechanisms mediate this exchange. We could calculate the influence of differential mutation rates, as well as that of biased transfer rates, on the partitioning of genes between the two genomes. We observe no significant difference in partitioning for haploid and diploid cell populations, but the effective size of cell populations is important. For infinitely large effective populations, higher mutation rates in mitochondria than in nuclear genomes are required to drive mitochondrial genes to the nuclear genome. In the more realistic case of finite populations, gene transfer favoring the nucleus and/or higher mutation rates in the mitochondrion will drive mitochondrial genes to the nucleus. We summarize experimental data that identify a gene transfer process mediated by vacuoles that favors the accumulation of mitochondrial genes in the nuclei of modern cells. Finally, we compare the behavior of mitochondrial genes for which transfer to the nucleus is neutral or influenced by purifying selection. PMID- 10833203 TI - Population structure and population history of Anopheles dirus mosquitoes in Southeast Asia. AB - Separating the confounding effects of long-term population history from gene flow can be difficult. Here, we address the question of what inferences about gene flow can be made from mitochondrial sequence data in three closely related species of mosquitoes, Anopheles dirus species A, C, and D, from southeast Asia. A total of 84 sequences of 923 bp of the mitochondrial cytochrome oxidase I gene were obtained from 14 populations in Thailand, Myanmar, and Bangladesh. The genealogy of sequences obtained from two populations of AN: dirus C indicates no contemporary gene flow between them. The F(ST) value of 0.421 therefore probably represents a recent common history, perhaps involving colonization events. Anopheles dirus A and D are parapatric, yet no differentiation was seen either within or between species. The starlike genealogy of their haplotypes, smooth unimodal mismatch distributions, and excess of low frequency mutations indicate population expansion in An. dirus A and D. This, rather than widespread gene flow, explains their low within-species F(ST) values (0.018 and 0.022). The greater genetic diversity of An. dirus D suggests that expansion occurred first in species D and subsequently in species A. The current geographical separation and low hybrid fitness of these species also argue against ongoing interspecific gene flow. They suggest instead either historical introgression of mtDNA from An. dirus D into species A followed by independent range expansions, or a selective sweep of mtDNA that originated in An. dirus D. While not excluding contemporary gene flow, historical population processes are sufficient to explain the data in An. dirus A and D. The genealogical relationships between haplotypes could not be used to make inferences of gene flow because of extensive homoplasy due to hypervariable sites and possibly also recombination. However, it is concluded that this approach, rather than the use of fixation indices, is required in the future to understand contemporary gene flow in these mosquitoes. The implications of these results for understanding gene flow in another important and comparable group of malaria vector mosquitoes in Africa, the An. gambiae complex, are also discussed. PMID- 10833204 TI - Statistical tests of gamma-distributed rate heterogeneity in models of sequence evolution in phylogenetics. PMID- 10833205 TI - Where do rodents fit? Evidence from the complete mitochondrial genome of Sciurus vulgaris. PMID- 10833206 TI - Master copy is not responsible for the high rate of copia transposition in Drosophila. PMID- 10833207 TI - Hemodynamic effects of sildenafil in men with severe coronary artery disease. AB - BACKGROUND: The cardiovascular effects of sildenafil are important because of the frequent presence of underlying cardiac disease in men with erectile dysfunction and reports indicating serious cardiac events temporally associated with the use of this drug. METHODS: We assessed the systemic, pulmonary, and coronary hemodynamic effects of oral sildenafil (100 mg) in 14 men (mean [+/-SD] age, 61+/ 11 years) with severe stenosis of at least one coronary artery (stenosis of >70 percent of the vessel diameter) who were scheduled to undergo percutaneous coronary revascularization. Blood-flow velocity and flow reserve were assessed with a Doppler guidewire in 25 coronary arteries, including 13 severely diseased arteries (mean stenosis, 78+/-7 percent) and 12 arteries without stenosis, used as a reference; maximal hyperemia was induced (to assess flow reserve) with the intracoronary administration of adenosine both before and after sildenafil. RESULTS: Oral sildenafil produced only small decreases (<10 percent) in systemic arterial and pulmonary arterial pressures, and it had no effect on pulmonary capillary wedge pressure, right atrial pressure, heart rate, or cardiac output. There were no significant changes in average peak coronary flow velocity, coronary-artery diameter, volumetric coronary blood flow, or coronary vascular resistance. Coronary flow reserve at base line was lower in the stenosed arteries (1.26+/-0.26) than in the reference arteries (2.19+/-0.44) and increased about 13 percent in both groups of arteries combined after the administration of sildenafil (from 1.70+/-0.59 to 1.92+/-0.72, P=0.003). The ratio of coronary flow reserve in coronary arteries with stenosis to that in the reference arteries (0.57+/-0.14) was not affected by sildenafil. CONCLUSIONS: No adverse cardiovascular effects of oral sildenafil were detected in men with severe coronary artery disease. PMID- 10833208 TI - A comparison of methotrexate with placebo for the maintenance of remission in Crohn's disease. North American Crohn's Study Group Investigators. AB - BACKGROUND: Patients with Crohn's disease often have relapses. Better treatments are needed for the maintenance of remission. Although methotrexate is an effective short-term treatment for Crohn's disease, its role in maintaining remissions is not known. METHODS: We conducted a double-blind, placebo controlled, multicenter study of patients with chronically active Crohn's disease who had entered remission after 16 to 24 weeks of treatment with 25 mg of methotrexate given intramuscularly once weekly. Patients were randomly assigned to receive either methotrexate at a dose of 15 mg intramuscularly once weekly or placebo for 40 weeks. No other treatments for Crohn's disease were permitted. We compared the efficacy of treatment by analyzing the proportion of patients who remained in remission at week 40. Remission was defined as a score of 150 or less on the Crohn's Disease Activity Index. RESULTS: Forty patients received methotrexate, and 36 received placebo. At week 40, 26 patients (65 percent) were in remission in the methotrexate group, as compared with 14 (39 percent) in the placebo group (P=0.04; absolute reduction in the risk of relapse, 26.1 percent; 95 percent confidence interval, 4.4 percent to 47.8 percent). Fewer patients in the methotrexate group than in the placebo group required prednisone for relapse (11 of 40 [28 percent] vs. 21 of 36 [58 percent], P=0.01). None of the patients who received methotrexate had a severe adverse event; one patient in this group withdrew because of nausea. CONCLUSIONS: In patients with Crohn's disease who enter remission after treatment with methotrexate, a low dose of methotrexate maintains remission. PMID- 10833209 TI - A preliminary study of growth hormone therapy for Crohn's disease. AB - BACKGROUND: Crohn's disease is a chronic inflammatory disorder of the bowel. In a preliminary study, we evaluated whether the administration of growth hormone (somatropin) as well as a high-protein diet would ameliorate the symptoms of the disease. METHODS: We randomly assigned 37 adults with moderate-to-severe active Crohn's disease to four months of self-administered injections of growth hormone (loading dose, 5 mg per day subcutaneously for one week, followed by a maintenance dose of 1.5 mg per day) or placebo. We instructed all patients to increase their protein intake to at least 2 g per kilogram of body weight per day. Patients continued to be treated by their usual physicians and to receive other medications for Crohn's disease. The primary end point was the change in scores on the Crohn's Disease Activity Index from base line to month 4. Scores can range from 0 to 600, with higher scores indicating more disease activity. RESULTS: At base line, the mean (+/-SD) score on the Crohn's Disease Activity Index was somewhat higher among the 19 patients in the growth hormone group than among the 18 patients in the placebo group (287+/-134 vs. 213+/-120, P=0.09). Three patients in the placebo group withdrew before their first follow-up visit and were not included in the data analysis. At four months, the Crohn's Disease Activity Index score had decreased by a mean of 143+/-144 points in the growth hormone group, as compared with a decrease of 19+/-63 points in the placebo group (P=0.004). Side effects in the growth hormone group included edema (in 10 patients) and headache (in 5) and usually resolved within the first month of treatment. CONCLUSIONS: Our preliminary study suggests that growth hormone may be a beneficial treatment for patients with Crohn's disease. PMID- 10833210 TI - Images in clinical medicine. Oral manifestations of Crohn's disease. PMID- 10833212 TI - Pediatric surgery. First of two parts. PMID- 10833211 TI - Coverage by the news media of the benefits and risks of medications. AB - BACKGROUND: The news media are an important source of information about new medical treatments, but there is concern that some coverage may be inaccurate and overly enthusiastic. METHODS: We studied coverage by U.S. news media of the benefits and risks of three medications that are used to prevent major diseases. The medications were pravastatin, a cholesterol-lowering drug for the prevention of cardiovascular disease; alendronate, a bisphosphonate for the treatment and prevention of osteoporosis; and aspirin, which is used for the prevention of cardiovascular disease. We analyzed a systematic probability sample of 180 newspaper articles (60 for each drug) and 27 television reports that appeared between 1994 and 1998. RESULTS: Of the 207 stories, 83 (40 percent) did not report benefits quantitatively. Of the 124 that did, 103 (83 percent) reported relative benefits only, 3 (2 percent) absolute benefits only, and 18 (15 percent) both absolute and relative benefits. Of the 207 stories, 98 (47 percent) mentioned potential harm to patients, and only 63 (30 percent) mentioned costs. Of the 170 stories citing an expert or a scientific study, 85 (50 percent) cited at least one expert or study with a financial tie to a manufacturer of the drug that had been disclosed in the scientific literature. These ties were disclosed in only 33 (39 percent) of the 85 stories. CONCLUSIONS: News-media stories about medications may include inadequate or incomplete information about the benefits, risks, and costs of the drugs as well as the financial ties between study groups or experts and pharmaceutical manufacturers. PMID- 10833213 TI - Clinical problem-solving. Letting the patient off the hook. PMID- 10833214 TI - Patients' rights bills and other futile gestures. PMID- 10833215 TI - New therapeutic approaches to Crohn's disease. PMID- 10833216 TI - The prospects for red-cell substitutes. PMID- 10833217 TI - Medical journals and medical reporting. PMID- 10833223 TI - Bacterioplankton Production in Humic Lake Ortrasket in Relation to Input of Bacterial Cells and Input of Allochthonous Organic Carbon. AB - In order to compare riverine bacteria input with lake water bacterial production and grazing loss with output loss, a bacterial cell budget was constructed for humic Lake Ortrasket in northern Sweden. The riverine input of bacterial cells in 1997 represented 29% of the number of bacterial cells produced within the layer of the lake affected by inlet water. A large share of the in situ lake bacterial production was consumed by grazers, mainly flagellates, which stresses the importance of bacteria as energy mobilizers for the pelagic food web in the lake. The bacterial production in Lake Ortrasket, which is almost entirely dependent on humic material as an energy source, was clearly stimulated by high flow episodes which brought high amounts of little degraded material into the lake. During base flow condition the bacterial production in the inlet rivers was high, which led to an input of more degraded material to the lake. This material did not stimulate the lake bacterial production. Internal factors that determined the utilization of the allochthonous DOC in the lake were the retention time and the exposure to light and high temperatures. Thus, the potential for in situ production of bacteria in Lake Ortrasket was to a large extent a function of how precipitation and runoff conditions affected terrestrial losses and river transport of humic material. PMID- 10833224 TI - Interrelationships between Rates of Microbial Production, Exopolymer Production, Microbial Biomass, and Sediment Stability in Biofilms of Intertidal Sediments. AB - The upper few millimeters of intertidal sediment supports a varied biomass of microbial consortia and microphytobenthos. Many of these organisms release extracellular polymers into the surrounding sediment matrix that can result in sediment cohesion and the increased stability of the sediment. The relationship between the heterotrophic and autotrophic components of these biofilms is not well understood. A combination of mesocosm and field investigations were used to investigate the relationship between microbial production rate (algae and bacteria), the extracellular carbohydrates, biomass, and stability in conjunction with a variety of environmental factors. An inverse relationship was found between rates of algal production and sediment stability both in the field and in laboratory mesocosms, though the relationship was significant only in the field (P < 0.001). Stability of sediments increased with increasing bacterial production rate (P < 0.001). Positive correlations were found between sediment stability and a range of other variables, including algal biomass (P < 0.001), colloidal-S EPS (P < 0.001), colloidal-S carbohydrate (P < 0.01), colloidal-S EDTA (P < 0.01), and sediment water content (P < 0.001). Using the data acquired, a preliminary model was developed to predict changes in sediment stability. Chlorophyll a, water content, and colloidal-S EPS were found to be the most important predictors of stability in intact cores incubated under laboratory conditions. Differences observed in patterns of the surface (0-2 mm) distribution of colloidal-S carbohydrate and chlorophyll a when expressed on a dry weight or areal basis were attributed to effects of dewatering and concomitant changes in wet bulk density. The polymeric carbohydrate (colloidal-S EPS) component of the biofilms was not found to be a constant fraction of the colloidal-S carbohydrate extract, varying from 16 to 58%, and the percentage of polymer decreased logarithmically as chlorophyll a concentrations increased and the biofilms matured (P < 0.001). Changes in the relationships between these variables over the period of biofilm development and maturation highlight the difficulties in their use to predict sediment stability. Exopolymer concentrations were more closely correlated with algal biomass than with bacterial numbers. Rates of algal carbon fixation were considerably greater than those for bacteria, suggesting that the algae have a much greater potential for exopolymer production. It is suggested that the microphytobenthos secretions make a more important contribution to sediment stability. PMID- 10833225 TI - Microdiversity of Culturable Diazotrophs from the Rhizoplanes of the Salt Marsh Grasses Spartina alterniflora and Juncus roemerianus. AB - Salt marshes dominated by Spartina alterniflora (smooth cordgrass) are among the most productive ecosystems known, despite nitrogen limitation. Rhizoplane/rhizosphere diazotrophy (nitrogen fixation) serves as a significant source of combined nitrogen in these systems. Several recent studies have demonstrated remarkable physiological and phylogenetic macro- and microdiversity within this important functional group of organisms. However, the ecological significance of this diversity is presently unknown. The physiological characteristics of the culturable, oxygen-utilizing fraction of the rhizoplane diazotroph assemblages from Spartina alterniflora and from another salt marsh grass, the black needle rush Juncus roemerianus, were examined in combination with an assessment of the phylogenetic relatedness by whole genome DNA-DNA hybridization. Analysis of substrate utilization data permitted quantitative evaluation of fully cross-hybridizing strain groups and physiological clusters. Phylogenetically related strains, defined by DNA homology >/=90% relative to the positive control, displayed extensive physiological diversity. Seven bootstrap supported physiological clusters, composed largely of phylogenetically dissimilar strains, showed similar utilization patterns for at least one class of ecologically relevant substrates (carbohydrates, carboxylic acids, or amino acids). These diazotrophs appear to be physiologically adapted for utilization of specific substrates or classes of substrates, lending support to diazotrophic functional redundancy. Microenvironmental heterogeneity is credited for promoting this diversity by selecting for physiologically specialized diazotroph populations to occupy defined niches in situ. One outcome of this physiological diversity is maintenance of a crucial environmental function (nitrogen fixation) over a broad range of environmental conditions. PMID- 10833226 TI - A Burkholderia Strain Living Inside the Arbuscular Mycorrhizal Fungus Gigaspora margarita Possesses the vacB Gene, Which Is Involved in Host Cell Colonization by Bacteria. AB - The arbuscular mycorrhizal (AM) fungus Gigaspora margarita harbors a resident population of endosymbiontic Burkholderia in its cytoplasm. Nothing is known about the acquisition of such bacteria and about the molecular bases which allow colonization of the fungus. We wondered whether the intracellular Burkholderia strain possesses genetic determinants involved in colonization of a eukaryotic cell. Using degenerated oligonucleotide primers for vacB, a gene involved in host cell colonization by pathogenic bacteria, an 842 bp DNA fragment was cloned, sequenced, and identified as a part of the vacB gene in Burkholderia sp. The insert was used as a probe to screen a fungal library that, because of the presence of intracellular Burkholderia cells, was also representative of the bacterial genome. The complete nucleotide sequence of vacB and flanking genes was determined. The bacterial origin of this genomic region was established by PCR, using specific vacB primers on DNA from Gigasporaceae that did or did not contain cytoplasmic Burkholderia, as well as on DNA from other bacteria, including free living Burkholderia. We hypothesize that the vacB gene is part of a new genetic region acquired by a rhizospheric Burkholderia strain, which became able to establish a symbiotic interaction with the AM fungus G. margarita. PMID- 10833227 TI - Aerobic Denitrifiers Isolated from Diverse Natural and Managed Ecosystems. AB - Twenty-eight bacterial strains were isolated from an ecosystem adapted to fluctuating oxic-anoxic conditions. This ecosystem comprised a mixture of different natural and wastewater treatment environments. Among the 28 strains isolated, 10 exhibited aerobic denitrifying activity, i.e., co-respiration of oxygen and nitrate and simultaneous production of nitrite by 4 of them and of nitrogen gas by the remaining 6. Comparisons between the 16S rDNA sequences of the 10 strains showed that 3 of them were identical to M. aerodenitrificans, whereas RAPD profiles showed that the 3 strains were identical to each other but that they were different from M. aerodenitrificans. This implies that alternating aerobic-anoxic conditions allowed the isolation of a new strain of this aerobic denitrifier. Moreover, other denitrifying bacteria belonging to the genera Paracoccus, Thiobacillus, Enterobacter, Comamonas, and Sphingomonas were isolated in this way. These data imply that a wide variety of bacteria are able to carry out this type of metabolism. M. aerodenitrificans was also detected in methanogenic, denitrifying, nitrifying, phosphate removal, and activated sludge ecosystems by two-step PCR amplification. After 4 months of acclimation to oxic anoxic phases, the strain was also detected in a canal and in a pond. This suggests that there is no specific natural ecological niche for aerobic denitrifiers but, as soon as selective pressure such as alternating aeration conditions is applied, this metabolism is amplified. PMID- 10833228 TI - Enrichment of Geobacter Species in Response to Stimulation of Fe(III) Reduction in Sandy Aquifer Sediments. AB - Engineered stimulation of Fe(III) has been proposed as a strategy to enhance the immobilization of radioactive and toxic metals in metal-contaminated subsurface environments. Therefore, laboratory and field studies were conducted to determine which microbial populations would respond to stimulation of Fe(III) reduction in the sediments of sandy aquifers. In laboratory studies, the addition of either various organic electron donors or electron shuttle compounds stimulated Fe(III) reduction and resulted in Geobacter sequences becoming important constituents of the Bacterial 16S rDNA sequences that could be detected with PCR amplification and denaturing gradient gel electrophoresis (DGGE). Quantification of Geobacteraceae sequences with a PCR most-probable-number technique indicated that the extent to which numbers of Geobacter increased was related to the degree of stimulation of Fe(III) reduction. Geothrix species were also enriched in some instances, but were orders of magnitude less numerous than Geobacter species. Shewanella species were not detected, even when organic compounds known to be electron donors for Shewanella species were used to stimulate Fe(III) reduction in the sediments. Geobacter species were also enriched in two field experiments in which Fe(III) reduction was stimulated with the addition of benzoate or aromatic hydrocarbons. The apparent growth of Geobacter species concurrent with increased Fe(III) reduction suggests that Geobacter species were responsible for much of the Fe(III) reduction in all of the stimulation approaches evaluated in three geographically distinct aquifers. Therefore, strategies for subsurface remediation that involve enhancing the activity of indigenous Fe(III)-reducing populations in aquifers should consider the physiological properties of Geobacter species in their treatment design. PMID- 10833229 TI - Reduced Growth Yield of Activated Sludge in Organic Protonophore-Containing Batch Culture. AB - The effects of organic protonophores 2,4-dinitrophenol (dNP) and para-nitrophenol (pNP) on the observed growth yield (Y(obs)) was studied using batch cultures of activated sludge microorganisms. A growth yield model was proposed in relation to the ratio of initial protonophore concentration (C(u)) to initial biomass concentration (X(0)) and was verified with experimental data. It was found that Y(obs) decreased with the increase of the C(u)/X(0) ratio, while the specific rate of glucose consumption was increased. Results showed that the C(u)/X(0) ratio could more reasonably reflect the real strength of organic protonophore exerted to activated sludge than using C(u) only. Based on the concept of growth yield, a model describing the uncoupling degree between energy generated via electron transport system and energy used for growth was further developed for protonophore-containing batch culture. It was shown that more than 60% of glucose was consumed through a futile cycle of energy rather than for growth at higher C(u)/X(0) ratios. This research usefully shows that the dissipation of energy via uncoupling biochemical processes can reduce excessive production of activated sludge markedly. PMID- 10833230 TI - Improving the aesthetics of the cheek bone projection during facial lifting. AB - This paper shows how one of the flaps of the SMAS, which is normally eliminated during a facelift as excess tissue, can be folded up and used to improve the projection of cheekbone. The improvement of the projection of this facial area enhances the aesthetic result, when the desired aesthetic result is discreet, in rhytidectomy without having to resort to biomedical material or complex surgical techniques. Our technique, which is safe and easy to handle, simplifies this procedure, providing a long-lasting satisfactory result for both the patient and the surgeon. Our report was done following rhytidectomy on 126 female patients and after an approximately 3-year follow-up. PMID- 10833231 TI - Our experience with endoscopic brow lifts. AB - This is a retrospective review of our experience with the endoscopic brow lift. We reviewed 128 procedures performed by two senior faculty members over the last 5 years. We evaluated the age, gender, operating time, complications, and outcome and conclude that endoscopic brow lift is a safe, efficient procedure with a low complication rate. The operating time is short, and there is a very high patient acceptance. The procedure has taken its place as an integral part of facial rejuvenation in our practice. PMID- 10833232 TI - Observations on a new concept for correction of the nasolabial fold in rhytidectomy. AB - Correction of the nasolabial fold has been the challenge constantly faced by the plastic surgeon since the start of attempts at facial rejuvenation. In this paper we intend to show observations which were made in a quite different method of approach to the correction of this challenging anatomic detail. It consists of taking the subcutaneous tissue which forms the nasolabial fold, underneath the same fold, following the law of gravity, without touching or dissecting the superficial aponeurotic muscular system (SMAS) and without using any other artifice which could make it difficult to observe this different procedure. The technique was used on 15 patients who requested facial rhytidectomy. PMID- 10833233 TI - Biomechanics and classification of the cartilaginous structures to project the nasal tip. AB - The present paper is the long-term conclusion of our preliminary presentation at the 1992 ISAPS Congress (Guadalajara, Mexico). This is the result of 29 cadaver dissections of different ages and both sexes. We have observed that length, thickness, and resistance correlate with the possibility to project the nasal tip with the mere structure of the alar cartilage and its medial crurae. We also demonstrated the existence and antagonistic action of Pitanguy's ligament as well as the depressing ligament to project the nasal tip. In this study, besides focusing on classifying the medial crus according to its thickness, length, and resistance, which is already different from any previous classification, we also focused on its surgical utility, and its interaction with other anatomic elements, to achieve the desired projection. Clinically, we present a 12-year experience with 1653 cases operated under this premise. PMID- 10833234 TI - Reed B. Bontecou, M.D.--his role in Civil War surgery and medical photography. AB - This third article on the subject of Civil War injuries and their surgical repair is devoted to Reed B. Bontecou, M.D., a New York surgeon who contributed greatly to the use of photography to document the casualties of the battlefield as seen in the Northern as well as the Southern states. Photographs of the wounded soldiers helped to verify the severity of their injuries, and helped to determine the degree and amount of the post-war pension payments. These photographs were one of the largest sources which were used in the creation of the Otis Archives of the present-day National Museum of Health and Medicine in Washington, D.C. Some of the wounded soldiers' photographs in the Otis Archives demonstrate the use of plastic surgery techniques to repair the wounded face, head and neck, torso, and the extremities by only a handful of Northern and Confederate surgeons including Gurdon Buck, J.S. Gouley and Henry B. Sands of New York City; H. Culbertson, U.S.V. of Madison, Wisconsin; J. Cooper McKee of Washington, D.C., W.W. Keen, Jr. of Philadelphia; and C.B. Gibson, P.A.C.S. of Richmond; S.H. Stout of Tennessee; and J.B. Bean, a dentist of Chattanooga. PMID- 10833235 TI - Aesthetic results of treatment of large alopecia with total scalp expansion. AB - In the last 7 years, 18 patients with large alopecia (approximately one-third of hair-bearing scalp) were treated with 31 tissue expansion procedures. In the previous 44 patients, with various degrees of alopecia treated with conventional tissue expansion technique, the major complication rates were as high as reported in the relevant literature. The clinical experience gained with these cases led us to make some modifications in the surgical technique. We applied some simple surgical maneuvers and Z-plasties to the last 18 cases with large alopecia. The major complication rate of 3.2% observed in this study is comparable to the lower complication rates of 6-12% reported in the literature for different degrees of alopecia. We believe that these simple modifications and meticulous approach improve the results of conventional treatment of alopecia with tissue expansion and flap. In this paper, details of the insertion technique and reconstructive procedures and their relevance to the success rate of the aesthetic treatment of the large alopecia are discussed. PMID- 10833236 TI - Reconstruction of scalp defects using simple designed bilobed flap. AB - We have devised a novel rational method to design a bilobed flap. In our method, two triangle flaps are designed; the angle of the first flap is three-fourths the angle of a rhombus at the defect site, and the angle of the second flap is also three-fourths that of the first flap. We have successfully performed reconstruction of scalp defects as large as 50 x 45 mm using simple designed bilobed flaps. The location of the defect was the parietal region in 10 cases, the frontal region in 6 cases, the temporal region in 3 cases, and the occipital region in one case. The advantages of this method for use in reconstruction of scalp defects are as follows: (1) the dispersion tension on the flap provides stable circulation and prevents expansion of scar formation; (2) since the suture line is zigzag, alopecia at the suture line can be hidden under the hair; (3) since this flap is a random pattern flap using galea aponeurotica with blood flow to the skin, flap design is possible for any part of the scalp; and (4) design and elevation of this flap are easy and do not take much time. PMID- 10833237 TI - Calfplasty. AB - The author describes a method of calf augmentation by a silicone prosthesis implant for correction of asymmetrical legs or aesthetic problems. Preference is given to the placement of soft silicone implants (Glitzentein implants); they are more natural looking than hard implants. The implants are placed over both heads of the gastrocnemius muscle or beneath the fascia cruris superficialis. Operations were performed from November 1985 to August 1999. One hundred patients were studied: 95 female and 5 male. Two hundred calf implants were performed: 188 soft implants (Glitzenstein), 6 hard implants (Aiache), and 6 together on the same leg (Glitzenstein and Aiache implants). No infection problems, prosthesis rupture, or hematomas occurred. There were four seromas, but they disappeared in 15 days. Four prostheses were removed in two patients who did not like the final result. One prosthesis changed place, to the upper part of the calf. The operation is performed under sedative and local anesthesia, with an incision of approximately 4 to 5 cm in the popliteal pleat, at the same height as the fascia cruris. To date, no functional problem or muscular dysfunction has been provoked by silicone prosthesis implants. The patient walks 8 h after the surgery. We selected 500 patients who received implants in the legs with calf prostheses, fat implants, and prosthesis and fat implant together, during 14 years of follow-up, but the focus of this paper is a calf implant with prosthesis in 100 patients with 200 calf prosthesis implants. PMID- 10833238 TI - Simultaneous breast augmentation and lift. AB - Often, both augmentation and mastopexy are necessary to solve the problems of breast ptosis with hypoplasia. These two procedures can be done simultaneously with no increased risks. Patients who have any degree of ptosis may benefit from some lifting of the nipple areola complex if the nipple is not in the central portion of the general contour of the breast mound when seen in the upright position. A simple crescent or eccentric excision in the upper quadrant may be sufficient to lift the nipple-areola complex 1-2 cm. If the nipple needs to be moved more than a couple of centimeters, or if the distance between the nipple and the inframammary crease is already excessive, an inframammary skin excision and redraping will be necessary. We have been using these combined techniques for 20 years with universal patient satisfaction. PMID- 10833239 TI - The thigh's lateral skin (saddle bags): histomorphometric study of interest to liposuction. AB - Study of the thigh's lateral skin ("saddle bags") in cadavers made it possible to evaluate its regional characteristics. Sex or race were not reported but the age group most appropriate to liposuction was considered. In this way, authors believed that as per standards established at the time, results as to the skin retraction in liposuction, will be able to be analyzed in a comparative way in the future. PMID- 10833241 TI - Surgeon scientist. AB - The origins and development of the renal transplant program at the Peter Bent Brigham Hospital (now the Brigham and Women's Hospital) from the late 1940s to the present are reviewed. The program was initiated as a effort to understand hypertension as a cause of renal failure. The initial transplants were unmodified allogeneic grafts placed in the thigh, followed by extensive laboratory experiments on dogs. This research culminated with the first successful human transplant of a kidney between identical twins in 1954. In 1959 the first successful fraternal allogeneic graft was accomplished as part of a protocol utilizing total body irradiation and bone marrow replacement. Finally, with the development of immunosuppressive drugs, we were able to transplant a cadaveric kidney successfully in 1962. This was a major impetus in the study of organ transplantation worldwide, which currently involves kidneys, liver, heart, pancreas, heart/lung, and bone marrow. PMID- 10833243 TI - Immunosuppression-back to the future. AB - During the 1950s work on bone marrow transplantation for patients with aplastic anemia led to the search for means other than total body irradiation to prevent rejection of the bone marrow allograft. At the same time, it had become clear that lymphocyte proliferation was a prominent feature of the immune response against all kinds of antigens, including skin allografts. These two factors led us to test various chemical agents known to block the proliferation of leukemic lymphocytes for their ability to inhibit the immune response against soluble antigens and skin allografts. One compound, the antileukemic drug 6 mercaptopurine, was effective in both test systems. This drug and its sister compound azathioprine (which is mercaptopurine with an imidizaole ring instead of a sulfhydryl group on carbon 6) were soon applied to human allografting and the treatment of autoimmune diseases. PMID- 10833244 TI - Tolerance and its clinical significance. AB - Several aspects of fetally or neonatally induced tolerance are briefly considered, and a variety of strategies for the induction of specific tolerance in adult animals or patients are discussed. They include the following: (1) augmentation of organ transplantation with donor-specific bone marrow; (2) the "prope" tolerance postulated by R.Y. Calne following treatment of kidney transplant recipients with a powerful monoclonal antibody; (3) linked epitope suppression, a form of specific tolerance for fully allogeneic haplotypes that can be induced in adult animals with a single major histocompatibility complex (MHC) epitope provided it is presented on the same antigen-presenting cells as the other MHC antigens; (4) allogeneic mixed chimerism, induced in experimental animals by the inoculation of allogeneic bone marrow into adult animals that had been subjected to a nonmyelosuppressive regimen, avoiding whole body irradiation; (5) several other approaches, viz., total lymphoid irradiation with bone marrow infusion, intrathymically induced tolerance, and the still rather mysterious case of the maternally noninherited HLA antigens of siblings. All or some of these strategies may yet come into their own in clinical transplantation, as the induction of specific tolerance must still be regarded as the most satisfactory solution to the problems of allogeneic, and perhaps xenogeneic, transplantation. PMID- 10833245 TI - Prope tolerance: a step in the search for tolerance in the clinic. AB - Following the demonstration by Knechtle and colleagues that profound T cell depletion in rhesus monkeys treated with a CD3 diphtheria immunotoxin resulted in tolerance to renal allografting, we have used a similarly depleting protocol that, in addition, depletes B cells in recipients of a cadaveric renal transplant. The humanized antiCD52 monoclonal antibody had a rapid depleting effect when given intravenously, 20 mg on days 0 and 1 after-renal transplantation, to 31 patients. At 48 hours after the second dose of low dose monotherapy cyclosporine (Neoral) was given to maintain blood levels averaging 100 ng/ml. Initially no other immunosuppression was given. With an average follow up of 24 months all but one of the patients are alive, 29 with intact functioning grafts. There have been five rejection episodes, which responded to pulsed steroid treatment. One patient had a recurrence of her original disease. Two patients developed opportunistic infections, which responded to therapy; and one patient with severe heart failure at the time of surgery died from this condition after 11 months. Currently 29 patients are still on the original low dose cyclosporine monotherapy. The outcome in this cohort of patients has been encouraging, with efficacy that compares favorably to our conventional triple therapy but in most cases allows the patients to be steroid-free on low dose immunosuppressive monotherapy. The maintenance treatment is inexpensive and should be beneficial in the context of tight budgetary constraints worldwide. The patients who avoided steroids are pleased with this aspect of the protocol. A randomized trial comparing this treatment with standard therapy is planned. PMID- 10833246 TI - Progress toward production of immunologic tolerance with no or minimal toxic immunosuppression for prevention of immunodeficiency and autoimmune diseases. AB - With donor and recipient matched at the major histocompatibility complex (MHC) locus, peripheral lymphoid tissue transplantation can be carried out without producing a graft-versus-host reaction or graft-versus-host disease (GVHD), thus correcting profound T cell immunodeficiencies of neonatally thymectomized mice. This analysis set the stage for clinical application of bone marrow transplantation (BMT) to provide for the first time cure of a human disease. With successful BMT, we cured immunologic deficiencies of a patient with XL severe combined immunodeficiency; thereafter we were the first to employ BMT to cure aplastic anemia. BMT regularly corrects immune and hematologic deficiencies caused by fatal irradiation without producing GVHD if the bone marrow (BM) used for the transplants has been purged of postthymic T cells. Over two decades in conjunction with Ikehara et al., we have shown that lethal total body irradiation (TBI) plus allogeneic BMT prevents or cures many organ-specific and systemic experimental autoimmune diseases. Animal models successfully treated by BMT include type I diabetes in nonobese diabetes (NOD) mice, type II diabetes in insulin-insensitive, glucose intolerant, diabetes mellitus (KK/Ay) mice, and autoimmune lupus erythematosus (LE) and glomerulonephritis in New Zealand Black x New Zealand White first generation hybrid (NZB x NZW)F1 females. El-Badri extended Ildstad's original research showing a high frequency of survival with a normal functioning immune system after stable mixed chimerism is produced by mixed BMT in C57BL/6 (normal long-lived black strain) mice transplanted with T cell-depleted marrow (TCDM) from BALB/c ("normal" long-lived strain) allogeneic donors and C57BL/6 syngeneic donors. We showed that osteoblasts act as facilitator cells for allogeneic BMT and promote engraftment of allogeneic hematopoietic stem cells. Wang et al. then showed that the autoimmunities and fulminating renal disease of BXSB (C57BL x SB cross and selective lupus-like systemic autoimmunity) male mice was prevented and could be cured by transplantation using TCDM from both BALB/c (resistant) and BXSB (susceptible) strains given to BXSB recipients after lethal TBI. This treatment produced mixed BMT and a stable mixed chimerism, increased longevity, corrected all manifestations of autoimmunity, and cured fulminant glomerulonephritis in these recipients. These studies generated a new perspective on the potential usefulness of BM and stem cell transplants to cure major diseases that can possibly be treated by BMT. Mixed BMT from TCD BALB/c and BXSB mice cured autoimmunities and fulminant glomerulonephritis in BXSB mice. LE disease plus coronary vascular disease that occurs in (NZW x BXSB)F1 mice, along with idiopathic thrombocytopenic purpura, is also cured in lethally irradiated (NZW x BXSB)F1 mice by BMT from C57BL/6 donors. Furthermore, hemolytic anemia, autoimmune phenomena, and hyalinizing glomerular renal disease of New Zealand Black (NZB) mice were prevented or cured by stem cell transplants using purified stem cells from MHC-matched DBA/2 donors or NZB donors. Consequently, we reasoned that autoimmunities reside in stem cells. More recently, we found that transplants of both BM cells and bones can completely and permanently prevent otherwise highly resistant autoimmune diseases of MRL/lpr lpr mice and an autoimmune polyarthritis of NZB/Kn mice. Ildstad concluded that lethal preparative measures would not be acceptable for preparations to treat autoimmune diseases, so we now employ a gentle method for producing stable mixed chimerism described by Sharabi and Sachs to achieve mixed marrow transplantation and mixed hematopoietic chimerism. Other diseases we are approaching using this gentle manipulation include two forms of diabetes: insulin-dependent diabetes mellitus (IDDM) type I in NOD mice and non insulin-dependent diabetes mellitus (NIDDM) type II in KK/Ay mice, atherosclerosis of apolipoprotein-E + kno PMID- 10833247 TI - Thoracic transplantation. AB - Experimental orthotopic transplantation of the heart was accomplished in 1959. Long-term survival was achieved in 1965 with a chemical immunosuppression protocol substantially different from that used for renal and hepatic transplants. Performance characteristics of the transplanted denervated heart were found to differ only slightly from normal. It appeared by the time of the Clinical Congress of the American College of Surgeons in October 1967 that clinical heart transplantation might be justified if the concept of brain death could be legally recognized. The Stanford program in clinical heart transplantation was inaugurated on January 6, 1968 and has been in continuous operation. To date, more than 1000 patients have undergone transplantation of the heart with the 5-year survival at 75%. The first long-term success in lung transplantation occurred at Stanford in 1981, with transplantation of the heart and both lungs. In 1990 the concept of living pulmonary lobar donors was introduced and is slowly finding its clinical role. The steroid-sparing capability of cyclosporine made possible both successful lung and pediatric heart transplantation. Only the donor shortage remains as a substantial barrier to widespread thoracic transplantation. Xenotransplantation is under intense scrutiny, with some encouraging experimental results. Development of the artificial heart continues to offer some relief for patients with end-stage heart disease. PMID- 10833248 TI - Landmarks in the development of hematopoietic cell transplantation. AB - The field of bone marrow transplantation has evolved over a period of 50 years. Reports of beneficial treatment of murine leukemia by irradiation and injection of marrow cells from another mouse stimulated interest in attempting to use these techniques to treat patients with leukemia. The first few bold attempts at human application were generally met with a total lack of success except for a few transplants involving identical twins. Understanding the HLA system led to the ability to select compatible sibling donors. The first successful long-term survivors were reported at the end of the 1960s. During the 1970's patients were given transplants for leukemia after failure of all other treatment. Survivals were poor, but some patients were cured. Transplantation early in the course of the disease resulted in greatly improved survival. During the 1980s improved control of infections, the use of peripheral blood as the source of stem cells, and the ability to select donors other than family members resulted in wide application of hematopoietic cell grafting. PMID- 10833249 TI - Human leukocyte antigen-G: immunotolerant major histocompatibility complex molecule in transplantation. AB - HLA-G is a nonclassical major histocompatibility complex class I molecule selectively expressed on cytotrophoblasts at the fetal-maternal interface, where it plays a role in maternofetal tolerance. In this review, attempts were made to summarize the current state of knowledge of the effects of HLA-G on both natural killer cell and T cell functions and their implications in transplantation. PMID- 10833250 TI - Double role of HLA in organ transplantation. AB - Although HLA matching can improve organ graft survival, well matched grafts fail and poorly matched ones survive. Recently evidence has been found that perinatal exposure to noninherited antigens (NIMAs) has a lifelong impact on the immune repertoire of the individual. The state of the art of the influence of NIMAs on graft survival is summarized. These findings could lead to a better understanding of the allograft reaction. PMID- 10833251 TI - Longmire lecture: my 50 years at the University of California, Los Angeles. AB - The main concept driving my work has been the humoral theory of immunity to allografts. It led to the development of the microlymphocytotoxicity test, which is used to look for the relevant transplant antigens using alloantisera. Using alloantibodies produced by pregnancies, the HLA system was defined through a series of international histocompatibility workshops. It was then shown that the HLA system was important for matching donors and recipients for bone marrow transplants and organ transplants. More than 6000 HLA-matched kidney transplants from cadaver donors have now been shared in the United States. HLA antigens were found to be of importance in anthropologic and disease susceptibility studies. Currently HLA antibodies are being studied intensively to determine their role in chronic rejection. If it is proven that these antibodies trigger intimal proliferation, occluding arterioles, HLA antibodies will become essential to the monitoring of chronic rejection. PMID- 10833242 TI - History of clinical transplantation. AB - The emergence of transplantation has seen the development of increasingly potent immunosuppressive agents, progressively better methods of tissue and organ preservation, refinements in histocompatibility matching, and numerous innovations in surgical techniques. Such efforts in combination ultimately made it possible to successfully engraft all of the organs and bone marrow cells in humans. At a more fundamental level, however, the transplantation enterprise hinged on two seminal turning points. The first was the recognition by Billingham, Brent, and Medawar in 1953 that it was possible to induce chimerism associated neonatal tolerance deliberately. This discovery escalated over the next 15 years to the first successful bone marrow transplantations in humans in 1968. The second turning point was the demonstration during the early 1960s that canine and human organ allografts could self-induce tolerance with the aid of immunosuppression. By the end of 1962, however, it had been incorrectly concluded that turning points one and two involved different immune mechanisms. The error was not corrected until well into the 1990s. In this historical account, the vast literature that sprang up during the intervening 30 years has been summarized. Although admirably documenting empiric progress in clinical transplantation, its failure to explain organ allograft acceptance predestined organ recipients to lifetime immunosuppression and precluded fundamental changes in the treatment policies. After it was discovered in 1992 that long-surviving organ transplant recipients had persistent microchimerism, it was possible to see the mechanistic commonality of organ and bone marrow transplantation. A clarifying central principle of immunology could then be synthesized with which to guide efforts to induce tolerance systematically to human tissues and perhaps ultimately to xenografts. PMID- 10833253 TI - Efficacy and safety of preoperative lobar or segmental ablation via transarterial administration of ethiodol and ethanol mixture for treatment of hepatocellular carcinoma: clinical study. AB - Transarterial embolization (TAE) using various thrombotic substances for unresectable hepatocellular carcinoma (HCC) performed on many patients has resulted in a better survival rate. We evaluated the efficacy and clinical safety of using an Ethiodol-ethanol mixture as the embolizer for treatment of HCC and the possibility of a surgical approach for inoperable tumors after TAE. Twenty patients with HCC who underwent TAE and tumor resection were included in the study. Initially, eight had increased retention rate of indocyanine green dye via intravenous injection (0.5 mg/kg) at 15 minutes (ICGR15), and six had an insufficient residual volume that precluded them from undergoing tumor resection. TAE was performed by slowly infusing the mixture of Ethiodol and ethanol into the artery supplying the tumor until dual hepatic artery and portal vein embolization was achieved. Serum levels of alanine aminotransferase increased after embolization, but all biochemistry studies reverted to normal within 2 weeks. A decreased tumor size (n = 15), improved ICG (n = 8), and increased volume of the nonembolized lobe (n = 10) were noted. The operations performed were right lobectomy (n = 11), extended right lobectomy (n = 3), left lobectomy (n = 2), extended left lobectomy (n = 2), and wedge resection (n = 2), which included patients who did not want to undergo major hepatectomy. Complete tumor necrosis was found in seven cases. All patients survived with no associated complications. The 1-year survival rate was 95%. Transarterial Ethiodol and ethanol administration creating dual hepatic artery and portal vein embolization was a safe and efficacious method for treating HCC. It effectively decreases tumor size, causes compensatory hepatic hypertrophy, and improves the ICGR15, which allows a wider range of patients to undergo liver surgery and achieve better survival. PMID- 10833252 TI - Historic landmarks in clinical transplantation: conclusions from the consensus conference at the University of California, Los Angeles. AB - The transplantation of organs, cells, and tissues has burgeoned during the last quarter century, with the development of multiple new specialty fields. However, the basic principles that made this possible were established over a three-decade period, beginning during World War II and ending in 1974. At the historical consensus conference held at UCLA in March 1999, 11 early workers in the basic science or clinical practice of transplantation (or both) reached agreement on the most significant contributions of this era that ultimately made transplantation the robust clinical discipline it is today. These discoveries and achievements are summarized here in six tables and annotated with references. PMID- 10833254 TI - Risk factors and true incidence of pouchitis in patients after ileal pouch-anal anastomoses. AB - Total colectomy, mucosal proctectomy, and ileal J pouch-anal anastomosis (IPAA) has become the procedure of choice for patients with ulcerative colitis and familial adenomatous polyposis. The purpose of this study was to determine the short- and long-term outcomes of patients undergoing IPAA by a single surgeon, correlating intraoperative technical aspects with outcomes, and to characterize better the clinical syndrome of pouchitis. A retrospective review was performed of 114 consecutive patients who underwent IPAA by a single surgeon between December 1987 and August 1996. Clinical follow-up and operative notes were reviewed, and patient questionnaires were obtained for all patients. The mean follow-up was 3 years (range 0.5-8.0 years). The average age of the patients was 39 years (range 16-72 years). There were 64 males and 50 females. Indications for operation were ulcerative colitis (n = 101) and familial polyposis coli (n = 13). Long-term morbidity occurred in 41% of patients (small bowel obstruction 10%, anastomotic stricture 9%). Pouch excision was required in only three patients. Stool frequency (mean +/- SE) was 6.1 +/- 0.2 and did not change with duration of follow-up. Only 7% of patients reported fecal soilage. The incidence of pouchitis was 59% (n = 67), with 4.2 +/- 0.3 episodes of pouchitis per patient. Using multivariate analysis, the factors significantly associated with the incidence of pouchitis were gender (p = 0.008) and duration of follow-up (p = 0. 02). A total of 37 of 50 women (74%) but only 30 of 64 men (47%) developed pouchitis. The incidence of pouchitis increased with the duration of follow-up. The incidences of pouchitis in patients followed for 6 months, 1 year, and 3 years were 25%, 37%, and 50%, respectively. Of patients followed more than 6 years, the incidence of pouchitis was 94% (15/16). There was not a significant correlation between anastomotic tension or the extent of arterial dissection of the ileal mesentery required to achieve IPAA and the incidence of pouchitis. The best antibiotics for pouchitis were metronidazole (54% of patients) and ciprofloxacin (37%). Eleven patients have required nearly continuous antibiotics. Patient satisfaction with the outcome is high, with a mean satisfaction of 8. 4 (0, dissatisfied; 10, extremely satisfied). This review demonstrates a high incidence of pouchitis in patients after IPAA, which is due to the more liberal definition of the syndrome and the complete follow-up achieved in this report compared to previous series. This study also is unique in identifying the significantly higher incidence of pouchitis in women, although the overall satisfaction with the clinical outcome in patients undergoing IPAA remains high. PMID- 10833255 TI - Primary gastric B cell lymphoma: audit of 82 cases treated with surgery and classified according to the concept of mucosa-associated lymphoid tissue lymphoma. AB - There has been a trend toward stomach conservation during treatment for primary gastric lymphoma, but surgery ensures accurate pathologic diagnosis through the examination of resected specimens. A histopathologic classification for primary gastric lymphoma had been established with the knowledge that the low grade and high grade components occasionally coexist in a single lesion. Efficacy of this classification as an indicator of biologic behavior and prognosis was evaluated. The classification was performed by reexamining surgically resected specimens from 82 patients with stage IE and IIE gastric lymphoma. Clinical characteristics of the low grade and high grade lymphomas were then evaluated and the survival data analyzed. Despite the larger tumor diameter, the depth of invasion for the low grade type was mostly submucosal, and none had serosal invasion. In contrast, 50% of the high grade type had serosal involvement and were also found to have a higher incidence of node metastasis. Prognosis of the low grade type, a 10-year survival rate exceeding 90%, was significantly better than that of the high grade type. Multivariate analysis revealed that curability of the operation followed by the histopathologic grade were significant independent prognostic factors. Some characteristics of the low grade lymphoma were not consistent with what has been reported in the studies involving stomach-conserving strategies. Investigators are advised to note that the histopathologic grade determined with biopsy specimens alone could be misleading. PMID- 10833256 TI - Effects of proximal duodenal transection and anastomosis on interdigestive sphincter of Oddi cyclic motility in conscious dogs. AB - Gallstones formed after gastrectomy are bilirubinate stones probably associated with biliary stasis and infection. Effects of proximal duodenal transection performed during gastrectomy on interdigestive sphincter of Oddi cyclic motility possibly relevant to this phenomenon were investigated in four conscious dogs. Although the cyclic change in sphincter motility was still in concert with the duodenal migrating motor complex after duodenal transection, the mean period was shortened (p < 0.02), and the frequency (p < 0.005) and amplitude (p < 0.001) of sphincter phasic waves during phase III were decreased. The cyclic variation of basal pressure disappeared, and the mean basal pressure throughout the cycle was significantly reduced (p < 0.003). Transient inhibition of sphincter and duodenal contractions normally seen during phase III disappeared. Duodenal transection reversed the response of the sphincter to cholecystokinin-octapeptide from inhibition to stimulation and from reduction of the basal pressure to elevation. These data suggest that duodenal transection produces significant changes in interdigestive sphincter of Oddi motility, possibly contributing to augmented duodenobiliary reflux and then lithogenesis. Myoneural continuity between the stomach and sphincter of Oddi at the proximal duodenum may play an important role in maintaining normal biliary dynamics. PMID- 10833257 TI - Manometric comprobation of esophagogastric junction competence after Nissen fundoplication and its relation to the length of fundic wrap. AB - We present 98 patients operated on by two techniques of 360-degree fundoplication due to gastroesopagheal reflux disease and divided in two groups: Group I comprised those who were given a conventional valve 5 cm in length, and group II underwent a short fundoplication that did not surpass 1.5 cm. All were subjected to preoperative esophageal manometry, with determination of the pressure and length of the lower esophageal sphincter (LES) and the motor behavior of the esophageal body. The postoperative manometric results are analyzed, checking that after fundoplication there was a significant increase in tone and length of the LES (p < 0.00005); there were no differences between the two groups (p = 0.9920 and p = 0.2160, respectively). The motor function of the esophageal body did not change after surgery. We conclude that even the smallest length of the fundic wrap does not cause a reduction in the pressure and length of the manometric LES; that is, the antireflux effectiveness, measured by manometry, did not depend on the anterior length of the valvuloplasty. PMID- 10833258 TI - Gastrointestinal anastomosis with the nickel-titanium double ring. AB - We used a dog model to test the safety and effectiveness of sutureless gastrointestinal anastomoses (n = 23) (end-to-end, end-to-side, or side-to-side) with the nickel-titanium double ring with temperature-dependent shape memory. Results were compared to conventional hand-sutured anastomoses (n = 5). Examination performed after 2 weeks' follow-up showed no leaks and no difference in return of bowel function between the experimental groups and the control group. Microscopic studies showed a complete epithelial lining and perfectly functioning anastomoses. We conclude that the nickel-titanium double-ring device may have an important place in the armamentarium of the gastrointestinal surgeon. PMID- 10833259 TI - The leech and the physician: biology, etymology, and medical practice with Hirudinea medicinalis. AB - The history of the word "leech" and the practice of leeching reveal interconnected social histories. We give the linguistic and medical histories of the word, and explore its biology and clinical history. Our historical account extends from the earliest known record of leeching to current research. Despite historical variation in its reputation as a therapeutic technique, leeching remains useful today in a number of applications. Further investigation may well disclose even more uses for the leech, particularly for its enzymes with anesthetic, anticoagulant, and antimetastatic properties. PMID- 10833260 TI - New crystal forms and low resolution structure analysis of 20S proteasomes from bovine liver. AB - 20S proteasomes from higher eukaryotes have immunological functions rather than those from archibacteria or yeast. To clarify the mechanism of the sorting and production of antigen-presenting peptides, it is important and worthwhile to determine the structure of mammalian proteasomes using a third generation synchrotron radiation source. Here we report new crystal forms of 20S proteasomes from bovine liver and preliminary structure analysis of them. The crystals belong to the same space group but have different cell dimensions. One crystal (form I) belongs to space group P2(1)2(1)2(1) with unit cell dimensions of a = 124.8, b =197.4, c =323.8 A, and diffracts to 3.0 A resolution. The other crystal (form II) belongs to the same space group with a =115.1, b =205.6, c =316. 0 A, and diffracts to 4.0 A resolution. The diffraction data for the form I crystal provided an interpretable electron density map for presenting the structural differences from yeast proteasomes. PMID- 10833261 TI - Backbone dynamics of the c-Myb DNA-binding domain complexed with a specific DNA. AB - The DNA-binding domain of c-Myb consists of three imperfect tandem repeats, R1, R2, and R3. Each repeat contains three helices. The minimal DNA-binding domain is an R2R3 fragment. Here, we have examined the backbone dynamics of R2R3 in its DNA bound form by NMR. Upon binding to DNA, the N- and C-termini, and the linker between R2 and R3 become less flexible. In the free form the third helix of R2 exhibits slow conformational exchange fluctuations owing to a cavity in the hydrophobic core of R2. Upon binding to DNA, the conformational exchange contributions in R2 are reduced but remain significant in NMR relaxation measurements. Upon binding to DNA, the third helix of R3 comes to exhibit significant chemical exchange contributions. These findings suggest that the orientations of the third helices of both R2 and R3 as to DNA are being chemically exchanged. In the DNA-bound form both R2 and R3 exhibit similar dynamical characters, except for amino acids Trp 95, Thr 96, and Val 103 of R2, which are located around the cavity of the unbound form. Upon binding to DNA, since Trp 95 moves into the cavity to fill it up, the local conformational exchange contributions seem to be still observable around the filled cavity. PMID- 10833263 TI - Studies on the phosphorylation of protein kinase B by Ca(2+)/calmodulin-dependent protein kinases. AB - Protein kinase B (PKB) was recently reported to be activated on the phosphorylation of Thr(308) by Ca(2+)/calmodulin-dependent protein kinase kinase alpha (CaM-kinase kinase alpha), suggesting that PKB was regulated through not only the phosphoinositide 3-kinase pathway but also the Ca(2+)/calmodulin protein kinase pathway. The activation of PKB by CaM-kinase kinase alpha was as high as 300-fold after incubation for 30 min under the phosphorylation conditions, and still increased thereafter, suggesting that the maximal activation of PKB on phosphorylation of the Thr(308) residue is several hundred fold. On the other hand, the V(max) value of CaM-kinase kinase alpha for the phosphorylation of PKB was more than two orders of magnitude lower than that for CaM-kinase IV, although the K(m) values for PKB and CaM-kinase IV were not significantly different, raising the question of whether or not PKB is a physiological substrate of CaM kinase kinase alpha. Besides CaM-kinase kinase alpha, CaM-kinase II also remarkably activated PKB. However, the specific activities of CaM-kinase kinase alpha and CaM-kinase II as to the activation of PKB were more than three orders of magnitude lower than that of 3-phosphoinositide-dependent protein kinase 1 (PDK1). PMID- 10833262 TI - Expression, purification, and characterization of blasticidin S deaminase (BSD) from Aspergillus terreus: the role of catalytic zinc in enzyme structure. AB - We established an efficient overproduction-purification system for blasticidin S deaminase (BSD) using the cDNA cloned from Aspergillus terreus. The estimated molecular mass of the purified enzyme indicated BSD was a tetramer. This tetrameric form was very resistant to denaturation by SDS and showed heat modifiable behavior on SDS-PAGE; i.e., BSD migrated much slower (as a single band of 36 kDa) in its active conformation than its completely denatured polypeptide (13 kDa) if heat treatment in 2% SDS was not performed before electrophoresis. As predicted from the presence of the catalytic zinc-coordinating sequence motif conserved in the cytosine nucleoside/nucleotide deaminase family, BSD also contained one zinc per deaminase subunit. However, the predicted catalytic function appeared not to be the only role of this zinc in the enzyme. First, titration of the zinc-chelating -SH groups with p-hydroxymercuriphenylsulfonate led to dissociation of the BSD tetramer into unstable monomers or dimers. Second, depletion of zinc on reconstitution of chemically denatured BSD (with either guanidine-HCl or acidic pH) resulted in improper folding of the polypeptide. These results suggest that zinc also plays a structural role in maintenance of the protein structure. When we introduced mutations at Glu-56 (the proposed active site) and Cys-91 (a proposed catalytic zinc-binding Cys) in BSD, none of the resulting mutants (E56D, E56Q, C91A, C91S, and C91H) showed any detectable activity, as judged with the spectrophotometric assay. Replacements of Cys-91 resulted in gross perturbation of the enzyme structure although the catalytically essential Glu-56 was not necessarily required for proper folding of the enzyme. These results further support our proposal that the catalytic zinc coordinated by the conserved sequence motif is also structural in BSD. PMID- 10833264 TI - Role of myeloperoxidase in the neutrophil-induced oxidation of low density lipoprotein as studied by myeloperoxidase-knockout mouse. AB - Low density lipoprotein was oxidized by neutrophils derived from either C57BL/6 mice or myeloperoxidase (MPO)-knockout mice. The generation of superoxide from neutrophils of MPO-knockout mice was about 70% of that from wild-type mice. The extent of the oxidation of human low density lipoprotein (LDL) by phorbol myristate acetate (PMA)-activated neutrophils of wild-type and MPO-knockout mice was assessed by measuring consumption of a-tocopherol and formation of phosphatidylcholine hydroperoxide (PCOOH) and cholesteryl ester hydroperoxide (CEOOH). Little consumption of a-tocopherol was observed in both oxidations. It was found, however, that lipid hydroperoxides were accumulated with time in both oxidations and that the rates of formation of PCOOH and CEOOH in the oxidation by MPO-knockout neutrophils were about 66 and 44% of those by wild-type neutrophils, respectively. The lipid peroxidation was completely inhibited by adding superoxide dismutase (SOD) in both cases. The addition of L-tyrosine and SOD enhanced lipid peroxidation of LDL induced by wild-type neutrophils but not by MPO-knockout ones. These results suggest that, regardless of their MPO activity, neutrophils induce lipid peroxidation of LDL by a superoxide-dependent pathway, and that MPO-catalyzed lipid peroxidation is enhanced by the presence of an appropriate amount of free tyrosine and further enhanced by SOD. PMID- 10833265 TI - Production of functional human selenocysteine-containing KDRF/thioredoxin reductase in E. coli. AB - In a previous study, we reported the isolation of a cDNA encoding KDRF (KM-102 derived reductase like factor) from the human bone marrow-derived stromal cell line KM-102. Analysis of the sequence of this cDNA revealed it to be the previously reported human thioredoxin reductase cDNA. Human thioredoxin reductase, which was recently isolated from human lung adenocarcinoma NCI-H441 cells as a selenocysteine-containing selenoprotein, and its substrate thioredoxin are thought to be essential for protecting cells from the damage caused by reactive oxygen species. To obtain the selenocysteine-containing recombinant KDRF/thioredoxin reductase, we introduced a secondary structure, which is identical to the selenocysteine insertion signal of Escherichia coli formate dehydrogenase H mRNA, downstream of the TGA in the KDRF/thioredoxin reductase cDNA and expressed it in E. coli. As a result, a significant amount of selenocysteine was incorporated into the C-terminus of the KDRF/thioredoxin reductase protein. The selenocysteine-containing KDRF/thioredoxin reductase showed reducing activities toward human and E. coli thioredoxin, whereas non selenocysteine-containing KDRF/thioredoxin reductase showed no enzyme activity. Our results suggest that this strategy will be applicable to the production of other mammalian selenocysteine-containing selenoproteins in E. coli. PMID- 10833266 TI - Purification of a low-molecular-weight phospholipase A(2) associated with soluble high-molecular-weight acidic proteins from rabbit nucleus pulposus and its comparison with a rabbit splenic group IIa phospholipase A(2). AB - An intervertebral disc is a large peice of avascular cartilage rich in proteoglycans and water consisting of gelatinous nucleus pulposus and fibrous annulus fibrosus. The soluble fraction of rabbit nucleus pulposus exhibited unusually high Ca(2+)-dependent phospholipase A(2) (PLA(2)) activity (about 70% of the total PLA(2) activity). The soluble PLA(2) activity was 6-7-fold higher than those of rabbit annulus fibrosus and spleen. The PLA(2) was bound to an anion-exchange column at pH 7.4, and eluted near the void volume as a broad peak on gel-filtration on a TSKgel SuperSW3000 column developed with a buffer containing 0.1-0.2 M salt. When the gel-filtration column was developed in the presence of 1 M salt, almost all the PLA(2) activity was eluted near the total available volume. The soluble PLA(2) was purified to near homogeneity. A Ca(2+) dependent PLA(2) was also purified from the fractions extracted with 1 M KBr from nucleus pulposus. For comparison, we purified a Ca(2+)-dependent PLA(2) from the KBr fraction of spleen. The splenic PLA(2) was identical to a group IIa PLA(2), as judged from its N-terminal amino acid sequences and mass spectra. On SDS polyacrylamide gel electrophoresis the enzymes purified from the soluble and KBr fractions of nucleus pulposus both gave a major 15. 7-kDa band at the same position as splenic group IIa PLA(2). These results suggest that group IIa PLA(2) is associated with soluble high-molecular-weight proteins, most likely proteoglycans, in the extracellular matrix of rabbit nucleus pulposus. PMID- 10833267 TI - Mutational analysis of the putative K(+)-binding site on the fourth transmembrane segment of the gastric H(+),K(+)-ATPase. AB - By means of a functional expression system and site-directed mutagenesis, we analyzed the role of the putative K(+)-binding site, Glu-345, located in the fourth transmembrane segment of the gastric H(+),K(+)-ATPase alpha-subunit. In the present study, we used several mutants, with alanine, isoleucine, leucine, glutamine, valine, lysine, and aspartic acid instead of Glu-345, and analyzed the H(+),K(+)-ATPase partial reactions of the mutants to determine the precise role of this residue. All the mutants except E345Q exhibited no H(+),K(+)-ATPase activity. The E345Q mutant showed 3-times higher affinity for ATP. This mutation shifted the optimum pH toward a more alkaline one. The E345A, E345I, E345L, E345V as well as E345Q mutants were phosphorylated with ATP as in the case of the wild type H(+),K(+)-ATPase, whereas the E345K mutant was not phosphorylated. The E345Q mutant was dephosphorylated in the presence of K(+), but its affinity for K(+) was significantly lower than that of the wild type. The E345A, E345I, E345L, and E345V mutants did not exhibit sensitivity to K(+) in the dephosphorylation step below 3 mM K(+). Therefore, Glu-345 is important for the conformational change induced by K(+), especially in the dephosphorylation step in which K(+) reacts with the enzyme from the luminal side with high affinity and accelerates the release of inorganic phosphate. The glutamic acid in the fourth transmembrane segment is conserved, and was found to be involved in the cation-induced conformational change in H(+),K(+)-ATPase as well as Na(+),K(+)-ATPase and Ca(2+) ATPase, however, the precise roles of the side chain in the function were different. PMID- 10833268 TI - Comparative study of the asparagine-linked sugar chains of human lipocalin-type prostaglandin D synthase purified from urine and amniotic fluid, and recombinantly expressed in Chinese hamster ovary cells. AB - Lipocalin-type prostaglandin D synthase (L-PGDS) is a highly glycosylated member of the lipocalin gene family and is secreted into various human body fluids. We comparatively analyzed the structures of asparagine-linked sugar chains of human L-PGDS produced by recombinant Chinese hamster ovary cells and naturally occurring human urine and amniotic fluid. After the sugar chains were liberated by hydrazinolysis followed by N-acetylation, they were derivatized with 2 aminobenzamide. All of the sugar chains of three L-PGDSs occur as biantennary complex-type sugar chains. Most of the sugar chains of three samples were fucosylated on the inner most N-acetylglucosamine residue. Although the sugar chains of the recombinant L-PGDS do not contain any bisecting N-acetylglucosamine residues, 58% and 34% of the fucosylated-sugar chains of amniotic fluid and urine L-PGDSs, respectively, contain bisecting N-acetylglucosamine residues. The sialic acid residues occur solely as Siaalpha2-->3Gal groups of the recombinant L-PGDS; the sialic acid residues of other L-PGDS occur as both Siaalpha2-->3Gal and Siaalpha2-->6Gal groups. Variations in L-PGDS glycosylation may prove useful as markers to further elucidate the role of L-PGDS glycoforms in different tissues. PMID- 10833269 TI - Free N-glycans already occur at an early stage of seed development. AB - As a part of our studies to elucidate the physiological significance of free N glycans in differentiating or growing plant cells, we first demonstrate that two kinds of free N-glycans already occur at an early stage of seed development. In this report, we used the developing Ginkgo biloba seeds as a model plant, since we have already revealed a functional feature of the Ginkgo endo-beta-N acetylglucosaminidase and structural features of N-glycans linked to storage glycoproteins in the developing seeds [Kimura, Y. et al. (1998) Biosci. Biotechnol. Biochem. 62, 253-261; Kimura, Y. and Matsuo, S. (2000) Biosci. Biotechnol. Biochem. 64, 562-568]. The structures of free N-glycans, which were determined by a combination of ESI-MS, sequential a-mannosidase digestions, partial acetolysis, and two dimensional sugar chain map, fell into two categories. One dominant species is a high-mannose type structure having one GlcNAc residue at the reducing end (Man(9-5)GlcNAc(1)). The concentration of this type of free glycan (as the pyridylaminated derivatives) is about 2.2 nmol in 1 g fresh weight. The detailed structural analysis revealed that the high-mannose type structures have a common core unit; Manalpha1-6(Man1-3)Manalpha1-6(Manalpha1 3)Ma nbeta1-4GlcNAc. The other minor species of free N-glycans is the plant complex type structure having an N-acetylchitobiose unit at the reducing end (Man(3)Xyl(1)Fuc(1)GlcNAc(2)). The concentration of this type of free glycan (as the pyridylaminated derivative) was about 75 pmol in 1 g fresh weight. PMID- 10833270 TI - Studies on the specificity and sensitivity of the influenza C virus binding assay for 9-O-acetylated sialic acids and its application to human melanomas. AB - The sensitivity and specificity of two influenza C virus assays, solid-phase and overlay assays, were investigated using naturally occurring 9-O-acetylated GD(3), rat serum glycoproteins containing 60% of N-acetyl-9-O-acetylneuraminic acid, and synthetically O-acetylated sialylated compounds. The sensitivity of the solid phase assay was higher for glycoproteins containing N-acetyl-9-O-acetylneuraminic acid than for gangliosides, and also differed for various 9-O-acetylated gangliosides. The overlay assay was less sensitive for all glycoconjugates tested. For virus recognition the presentation of the sialic acid within the molecule and the structure of the sialic acid are essential. Investigation of gangliosides from human melanomas and normal skin with the influenza C virus assay showed an increase of O-acetylation of sialic acids in most tumour samples and the occurrence of several O-acetylated gangliosides. PMID- 10833271 TI - Purification, characterization, and primary structure of a novel cell wall hydrolytic amidase, CwhA, from Achromobacter lyticus. AB - A novel bacteriolytic enzyme CwhA (cell wall hydrolytic amidase) was purified by ion exchange and gel-filtration chromatographies from a commercial bacteriolytic preparation from Achromobacter lyticus. CwhA exhibited optimal pH at 8.5 and lysed CHCl(3)-treated Escherichia coli more efficiently than Micrococcus luteus, Staphylococcus aureus, Enterococcus faecalis, and Pediococcus acidilactici. The enzyme was inhibited by 1,10-phenanthroline strongly and by EDTA to a lesser extent, suggesting that it is probably a metalloenzyme. Amino acid composition and mass spectrometric analyses for the CwhA-derived M. luteus muropeptides revealed that CwhA is N-acetylmuramoyl-L-alanine amidase [EC 3.5.1. 28]. The complete amino acid sequence of CwhA was established by a combination of Edman degradation and mass spectrometry for peptides obtained by Achromobacter protease I (API) digestion and cyanogen bromide (CNBr) cleavage. The enzyme consists of a single polypeptide chain of 177 amino acid residues with one disulfide bond, Cys114-Cys121. CwhA was found to be homologous to N-acetylmuramoyl-L-alanine amidase from bacteriophage T7 (BPT7). Its sequence identity with BPT7 is 35%, but the amino acid residues functioning as zinc ligands in BPT7 are absent in CwhA. These results suggest that CwhA is a new type of N-acetylmuramoyl-L-alanine amidase. PMID- 10833272 TI - Inhibitory effects of 1,4-naphthoquinone derivatives on rat cytochrome P4501A1 dependent monooxygenase activity in recombinant yeast microsomes. AB - We reported previously that various naphthoquinone derivatives inhibited cytochrome P450-dependent monooxygenase of liver and placenta microsomes [Muto, N. et al. (1987) Biochem. Biophys. Res. Commun. 146, 487-494]. To understand the complex inhibitory behaviors that were observed, it is desirable to study the relationship between structure and inhibitory activity of naphthoquinones in a simplified system containing a single P450 species. In the present study, the inhibitory effects of six derivatives of 1,4-naphthoquinone (hereafter referred to as NQ) on rat cytochrome P4501A1-dependent 7-ethoxycoumarin O-deethylation were examined using yeast microsomes containing overexpressed rat P4501A1. Of these, 2-methyl-5-hydroxy-NQ, 2-methyl-NQ, 2-hydroxy-NQ, and NQ showed competitive inhibition, whereas 5,8-dihydroxy-NQ and 5-hydroxy-NQ showed noncompetitive inhibition. Judging from the inhibitor constant (K(i)), the binding affinity of the four competitive inhibitors for the substrate-binding pocket of P4501A1 is in the order: 2-CH(3)-5-OH-NQ > 2-CH(3)-NQ > NQ >> 2-OH-NQ. On binding with P4501A1, 2-CH(3)-5-OH-NQ, 2-CH(3)-NQ, and NQ induced distinct Type II, Type I, and reverse Type I spectra, respectively. These results indicate that methyl and hydroxyl groups introduced into NQ have unique effects on their binding mode and binding affinity. PMID- 10833273 TI - Characterization of human liver leukotriene B(4) omega-hydroxylase P450 (CYP4F2). AB - We previously reported the cloning of a human liver leukotriene B(4) (LTB(4)) omega-hydroxylase P450 designated CYP 4F2 [Kikuta et al. (1994) FEBS Lett. 348, 70-74]. However, the properties of CYP 4F2 remain poorly defined. The preparation solubilized using n-octyl-beta-D-glucopyranoside from microsomes of CYP 4F2 expressing yeast cells catalyzes v- hydroxylation of LTB(4), 6-trans-LTB(4), lipoxin A(4), 8-hydroxyeicosatetraenoate, 12-hydroxyeicosatetraenoate, and 12 hydroxystearate in the presence of rabbit liver NADPH-P450 reductase. In addition, the enzyme shows ethoxycoumarin O-deethylase and p-nitroanisole O demethylase activities. The enzyme was purified to apparent electrophoretic homogeneity from yeast cells by sequential chromatography of solubilized microsomes through amino-n-hexyl-Sepharose 4B, DEAE-HPLC, and hydroxylapatite HPLC columns. The final preparation showed a specific content of 11.1 nmol of P450/mg of protein, with an apparent molecular mass of 56.3 kDa. CYP 4F2 was distinguished from the closely homologous CYP 4F3 (human neutrophil LTB(4) omega hydroxylase) by its much higher K(m) for LTB(4), inability to omega-hydroxylate lipoxin B(4), and extreme instability. PMID- 10833274 TI - 19F NMR investigation of F(1)-ATPase of Escherichia coli using fluorotryptophan labeling. AB - Growth of Escherichia coli in the presence of glyphosate, an inhibitor of aromatic amino acid biosynthesis, has permitted the production of proton dislocating ATPase that is specifically labeled with 5-fluorotryptophan. Five sets of (19)F resonances could be assigned to each tryptophan residue by lauryldimethylamine oxide and carboxypeptidase treatment. On labeling with 4 chloro-7-nitro-benzofurazan, the label attached to b155Lys, which is known to be in the catalytic site, which caused one of the residues, b108Trp, to become nonequivalent. (19)F NMR spectroscopic investigation of internally fluorotryptophan-labeled F(1)-ATPase will provide valuable information about the asymmetric nature of F(1)-ATPase and the conformational changes induced by ligand binding. PMID- 10833275 TI - Affinity selection of DNA-binding proteins displayed on bacteriophage lambda. AB - Two transcription factors, human ATF1, its DNA-binding domain (ATF1BD), and the DNA-binding domain (GAL4BD) of the yeast GAL4 protein, were displayed on the surface of bacteriophage lambda vectors and efficiently selected by DNA fragments immobilized in microtiter wells. The DNA-binding proteins are fused to the carboxy terminus of the tail protein gpV and head protein gpD of the vectors, lambdafoo and lambdafooDc, respectively. After a single round of affinity selection, the fusion phages were successfully enriched 60- to 4,000-fold over the vector phages. Further, the GAL4BD fusion phages were enriched 5- and 15-fold by affinity selection using specific DNA as probes over nonspecific DNA when expressed on lambdafooDc and lambdafoo, respectively. The ATF1BD fusion phages were also sequence-specifically enriched greater than 4-fold when displayed on lambdafoo. These results suggest that the lambdafoo display system is useful for in vitro studying of protein-DNA interactions and may be applied to screening of DNA-binding protein from complex cDNA libraries through DNA-binding affinity. PMID- 10833276 TI - Cloning and characterization of a myosin from characean alga, the fastest motor protein in the world. AB - In characean algae, very rapid cytoplasmic streaming is generated by sliding movement of an unconventional myosin on fixed actin cables. The speed of this sliding movement is the fastest among many molecular motors known so far. We have cloned a set of overlapping cDNAs encoding the heavy chain of this myosin by immunoscreening with antibody raised against characean myosin. The molecular mass of this heavy chain is 248 kDa, and the protein has a conserved motor domain, six IQ motifs, an extensive alpha-helical coiled-coil domain, and a C-terminal globular domain. Phylogenetic analysis suggested that this myosin belongs to class XI. PMID- 10833278 TI - Enzymatic removal of oxidized protein aggregates from erythrocyte membranes. AB - Erythrocytes oxidized or aged in the circulation undergo membrane protein aggregation and anti-band 3 autoantibody binding to the cell surface. When human erythrocytes were mildly oxidized in vitro with 0.1 mM Fe(III) at 37 degrees C for 3 h, the aggregation of nonionic detergent C(12)E(8)-insoluble membrane protein and the binding of anti-band 3 IgG to the cell surface were increased. Incubation of membranes isolated from the oxidized cells increased the amount of protein aggregates by 5-fold after 6 h, while incubation for a further 12 h sharply decreased the amount of aggregates. In the presence of diisopropyl fluorophosphate (DFP), however, the increased amount of aggregates was maintained in the subsequent incubation. Western blot analysis of the aggregates using rabbit anti-band 3 showed that band 3 protein aggregates increased in the initial stage of incubation and decreased upon subsequent incubation, whereas the increased band 3 protein aggregates did not subsequently decrease when membranes were incubated in the presence of DFP. Incubation of the oxidized cells at 37 degrees C for 18 h caused reduction of the membrane protein aggregates and the (125)I-anti-band 3 IgG binding to the cell surface, while incubation in the presence of DFP did not cause these reductions. The results suggest that the oxidation-induced cell membrane protein aggregates were probably removed by 80 kDa serine protease, namely, oxidized protein hydrolase (OPH), in the oxidized cell membranes [Fujino et al. (1998) Biochim. Biophys. Acta 1374, 47-54; (1998) J. Biochem. 124, 1077-1085; (2000) Biochim. Biophys. Acta 1478, 102-112], and as a result the increased anti-band 3 binding to the cell surface was reduced. PMID- 10833277 TI - Characterization of a mutant form of SecA that alleviates a SecY defect at low temperature and shows a synthetic defect with SecY alteration at high temperature. AB - The secY205 mutant is cold-sensitive for protein export, with an in vitro defect in supporting ATP- and preprotein-dependent insertion of SecA into the membrane. We characterized SecA81 with a Gly516 to Asp substitution near the minor ATP binding region, which suppresses the secY205 defect at low temperature and exhibits an allele-specific synthetic defect with the same SecY alteration at 42 degrees C. The overproduced SecA81 aggregated in vivo at temperatures above 37 degrees C. Purified SecA81 exhibited markedly enhanced intrinsic and membrane ATPase activities at 30 degrees C, while it was totally inactive at 42 degrees C. The trypsin digestion patterns indicated that SecA81 has some disorder in the central region of SecA, which encompasses residues 421-575. This conformational abnormality may result in unregulated ATPase at low temperature as well as the thermosensitivity of the mutant protein. In the presence of both proOmpA and the wild-type membrane vesicles, however, the thermosensitivity was alleviated, and SecA81 was able to catalyze significant levels of proOmpA-stimulated ATP hydrolysis as well as proOmpA translocation at 42 degrees C. While SecA81 was able to overcome the SecY205 defect at low temperature, the SecY205 membrane vesicles could not significantly support the translocation ATPase or the proOmpA translocation activity of SecA81 at 42 degrees C. The inactivated SecA81 molecules seemed to jam the translocase since it interfered with translocase functions at 42 degrees C. Based on these results, we propose that under preprotein-translocating conditions, the SecYEG channel can stabilize and activate SecA, and that this aspect is defective for the SecA81-SecY205 combination. The data also suggest that the conformation of the central region of SecA is important for the regulation of ATP hydrolysis and for the productive interaction of SecA with SecY. PMID- 10833279 TI - Identification of the cleavage sites of oxidized protein that are susceptible to oxidized protein hydrolase (OPH) in the primary and tertiary structures of the protein. AB - Amino acid sequences in H(2)O(2)-oxidized bovine serum albumin (BSA) that are susceptible to proteolytic cleavage by oxidized protein hydrolase (OPH) were investigated. When oxidized BSA was treated with OPH, low-molecular-weight fragments (54, 46, 24, 22, 20, and 8 kDa) were produced as analyzed by SDS-PAGE. N-Terminal amino acid sequence analysis of these fragments indicated that oxidized BSA was cleaved by OPH at three major sites, Leu218-Ser219, Tyr410 Thr411, and Phe506-Thr507, at an early stage of the proteolytic degradation. In the three-dimensional structure of BSA deduced by computer modeling, these cleavage sites were found to be located slightly inside the BSA molecule, in positions not easily accessible by OPH. The influence of oxidation on the tertiary structure of BSA was then investigated by hypothetically replacing all the four methionine and two tryptophan residues with their oxidized forms, methionine sulfoxide and N'-formyl-kynurenine, respectively. The three dimensional structure of the hypothetically oxidized BSA indicated that all the three cleavage sites in the protein could become more exposed to the solvent than in unoxidized BSA. These results suggest that, upon oxidation of BSA, the amino acid sequences that are potentially cleavable by OPH but present inside the molecule become exposed on the surface and susceptible to proteolysis by OPH. This is the first report demonstrating the cleavage sites of oxidized protein by oxidized protein-selective protease, suggesting the possible mechanism of oxidized protein-selective degradation by the enzyme. PMID- 10833280 TI - The effect of single residue substitutions of serine-283 on the strength of head to-tail interaction and actin binding properties of rabbit skeletal muscle alpha tropomyosin. AB - Vertebrate skeletal muscle alpha-tropomyosin polymerizes in a head-to-tail manner and binds cooperatively to actin. It has been postulated that the cooperative actin binding is governed by the strength of the head-to-tail interaction. In order to know the relationship between the head-to-tail affinity and actin binding, we studied the properties of tropomyosin variants with single residue substitutions at serine-283, the penultimate residue at the carboxyl terminus that is involved in the head-to-tail interaction. It has been shown that the phosphorylation of serine-283 strengthens the head-to-tail interaction. Viscometry was employed to compare the head-to-tail affinity of tropomyosin variants. Variant S283E showed higher viscosity whereas variant S283K showed lower viscosity compared with the wild type non-phosphorylated alpha-tropomyosin. The results confirm the idea that the interaction is sensitive to the ionic properties of residue 283. The strength of the head-to-tail interaction was assessed directly by sedimentation equilibrium using two pairs of tropomyosin variants designed so that only dimeric interactions were allowed within each pair. From one pair of variants with serine-283, the association constant was determined to be 2.6 x 10(4) M(-1) (SD =1.0 x 10(4)), whereas for the second pair with glutamate-283, the affinity was 3.9 x 10(4) M(-1) (SD =1.6 x 10(4)), slightly stronger than the former, consistent with the results of viscometry. The results indicate that the head-to-tail association is weak as previously implicated from light scattering measurements. Cosedimentation was employed to measure the cooperative actin binding of tropomyosin variants. Although previous results indicated the phosphorylation has no significant influence on the actin affinity, variant S283E shows a lower affinity compared with the control. Variants S283K and S283A show even lower affinities to actin, although these species bind to actin more cooperatively than does variant S283E. The results indicate that the affinity of the head-to-tail interaction between adjacent tropomyosin molecules is weak, and is substantially influenced by an extra charge at residue 283. On the other hand, the interaction with actin, the affinity and the cooperativity in actin binding, is dependent on amino acid residues at 283 and is not simply correlated with the strength of the head-to-tail interaction between Tm molecules in solution. PMID- 10833281 TI - The ratio of splicing variants of MGC-24/CD164, a sialomucin, correlates with the metastatic potential of colorectal carcinomas. AB - MGC-24/CD164 is a sialomucin expressed in many normal and cancerous tissues. In humans, soluble and transmembrane forms of MGC-24 are produced by alternative splicing. The total MGC-24 RNA level was found to be lower in human colorectal carcinomas as compared with the adjacent normal mucosal tissues. Lower MGC-24 mRNA levels in colon carcinomas and in the adjacent normal mucosa epithelium correlate with lymphatic vessel invasion by the carcinoma. The ratio of the soluble form to the transmembrane form of the mRNA in colorectal carcinomas was determined by ribonuclease protection assay. Higher ratios were correlated with less venous invasion and less remote metastasis, which became evident during postoperative observation. PMID- 10833282 TI - Characterization of recombinant human DNA topoisomerase IIIalpha activity expressed in yeast. AB - Recombinant human DNA topoisomerase IIIalpha was expressed in mutant yeast cells devoid of both topoisomerases I and III, and the gene product was partially purified. The activity of the protein in supercoil removal was found to be limited and also biphasic: in the first phase it processively changed the linking number of hypernegatively supercoiled DNA, but only to the superhelicity of a regular negative supercoil; in the second phase the enzyme relaxed the DNA further, but only slightly and slowly. The optimal solution conditions for the enzyme activity were found to be physiological. The assay results with a truncation mutant showed that the C-terminal 334 amino acids are unnecessary for the activity, suggesting that this region, and perhaps the entire protein, is involved in a function other than supercoil removal. PMID- 10833283 TI - Conformational changes of dynein: mapping and sequence analysis of ATP/Vanadate dependent trypsin-sensitive sites on the outer arm dynein b heavy chain from sea urchin sperm flagella. AB - Conformational changes of dynein during ATP hydrolysis are demonstrated by the difference in the tryptic fragments of the dynein heavy chain between in the absence and presence of ATP and vanadate. Here tryptic sites in the presence of ATP and vanadate (Tav sites) have been mapped on the betaheavy chain of outer arm dynein from sea urchin sperm flagella. Tav sites are located not only near the central catalytic domain which includes four P-loops, but also near the carboxyl terminal coiled-coil region. The Tav2 site is located in the most carboxyl terminal region, which is nearly 850 amino acid residues apart from the the fourth P-loop (P4 site). The region from the most amino-terminal Tav site (Tav1 site) to the Tav2 site covers approximately 2,100 amino acid residues, which is almost half the whole betaheavy chain. Comparison of the sequences around the tryptic sites of the sea urchin b chain and those of the dynein heavy chains from other organisms reveals that the sequence around the Tav1 site is highly conserved in both cytoplasmic and axonemal dyneins but that around Tav2 sites is only conserved in axonemal dyneins, suggesting functional differences in the Tav2 region between the two subfamilies of dynein. PMID- 10833284 TI - Processing pathway deduced from the structures of N-glycans in Carica papaya. AB - A processing The processing pathway of N-glycans in Carica papaya was deduced from the structures of N-glycans. The N-glycans were liberated by hydrazinolysis followed by N-acetylation. Their reducing-end sugar residues were tagged with 2 aminopyridine and the pyridylamino (PA-) sugar chains thus obtained were purified by HPLC. Eleven PA-sugar chains were found, and their structures were analyzed by two-dimensional sugar mapping combined with partial acid hydrolysis and exoglycosidase digestion. The structures of the N-glycans were of the highmannose types with xylose and fucose; however, among them two new N-glycans, Manalpha1 6(Manalpha1-3)Manalpha1-6(Xylbeta1-2)+ ++Manbeta1-4GlcNAcbeta1- 4(Fucalpha1 3)GlcNAc and Manalpha1-3Manalpha1-6(Xylbeta1-2)Manbeta1-4G lcNAcbeta1-4(Fucalpha1 3 )GlcNAc, were found. Judging from these structures together with Manalpha1 6(Manalpha1-3)Manalpha1-6(Manalpha1-3) (Xylbeta1-2)Manbeta1- 4GlcNAcbeta1 4(Fucalpha1-3)GlcNAc reported previously [Shimazaki, A., Makino, Y., Omichi, K., Odani, S., and Hase, S. (1999) J. Biochem. 125, 560- 565], a processing pathway for N-glycans in C. papaya is inferred in which the activity of Golgi alpha mannosidase II is incomplete. PMID- 10833285 TI - Molecular cloning and characterization of SRAM, a novel insect rel/ankyrin-family protein present in nuclei. AB - Previously, we purified a 59-kDa protein that binds to the kappaB motif of the Sarcophaga lectin gene. Here we report its cDNA cloning and some of its characteristics as a novel member of the Rel/Ankyrin-family. This protein, named SRAM, contained a Rel homology domain, a nuclear localization signal and 4 ankyrin repeats, but lacked the Ser-rich domain and PEST sequence that Relish contained. We found that SRAM was localized in the nuclei of NIH-Sape-4 cells, which are an embryonic cell line of Sarcophaga. The Sarcophaga lectin gene promoter containing tandem repeats of the kappaB motifs was activated in NIH-Sape 4 cells. In Drosophila mbn-2 cells, Dif alone activated this reporter gene and a cooperative effect was detected when SRAM and Dif were co-transfected, although SRAM alone did not activate it. This is the first report of a Rel/Ankyrin molecule that exists in the nuclei. PMID- 10833286 TI - The response of human buccal maxillary furcation defects to combined regenerative techniques--two controlled clinical studies. AB - Treatment of maxillary furcation lesions presents important clinical challenges. The aim of this study was to evaluate the response of buccal class II furcation defects in systemically healthy, non-smoking, Adult Periodontitis patients undergoing supportive periodontal therapy and surgical periodontal regenerative techniques. The treatment protocol emphasised infection control, careful soft tissue management and wound stability. Two experiments were performed. In the first experiment, a coronally positioned flap (CPF) technique was compared with surgical debridement. In the second experiment, an experimental technique (ET) was also compared with surgical debridement. The ET was a combination of a composite graft consisting of microparticulate (40-70 microns) hydroxyapatite and tetracycline (3:1), a guided tissue regeneration barrier (GTR) and CPF. Sixty patients were divided into the four (n = 15) treatment groups. The clinical variables evaluated were plaque, bleeding-on-probing scores, gingival recession, probing pocket depth (PD), vertical (VAL) and horizontal attachment levels (HAL). Reevaluation was performed 12 months after the surgical procedures and revealed that the three techniques resulted in improvements in all the clinical variables evaluated. Post-operative measurements from controls, CPF and ET showed (respectively) 1.53 +/- 1.12 mm, 1.40 +/- 0.75 mm, 2.43 +/- 1.36 mm reduction in PD; 0.63 +/- 0.1 mm, 1.17 +/- 1.12 mm and 1.57 +/- 1.32 mm VAL gains and 1.03 +/- 0.30 mm, 1.40 +/- 0.96 mm and 2.13 +/- 1.52 mm HAL gains. Five out of fifteen furcations in the ET group were closed after treatment. It was concluded that satisfactory clinical results--measured by PD reduction, VAL and HAL gains--can be obtained in advanced defects formerly considered as being non-responsive to regenerative periodontal therapy. The proposed ET showed significantly better results, with more PD reduction, HAL and VAL gains and a higher frequency of furcation closure compared with the other techniques tested, and shows promise as a new regenerative treatment technique. PMID- 10833287 TI - Helicobacter pylori in dental plaque of Pakistanis. AB - Helicobacter pylori is now generally accepted to play a key role in acid related and neoplastic pathology of gastroduodenal diseases. Recent reports have concluded that dental plaque is not an important reservoir for Helicobacter pylori, however, these studies did not consider the ethnic background of their subjects nor the amounts of dental plaque present. The aim of this study was to explore the association of Helicobacter pylori dental plaque colonisation in 125 males and 53 females (group I) attending a dental clinic in Pakistan. A simultaneous sample of 30 healthy volunteers with good orodental hygiene consisting of 17 males and 13 females was included as a control group (group II). Six dental plaque specimens were obtained from each subject with a sickle scaler; two were inoculated into CLO test gel and the remaining four were used to prepare cytology slides stained with Giemsa's stain. CLO test was positive in all specimens from group I, while cytology for Helicobacter pylori was positive in 173 cases in this group. One hundred and forty two cases had heavy plaque deposits and all of them were positive on cytology. In group II CLO test was positive in 20 and dental plaque cytology was positive in 7 cases. In conclusion, it is important that future studies into the prevalence of Helicobacter pylori in the oral cavity should take into account the levels of oral cleanliness and the ethnic background of the subjects. PMID- 10833288 TI - Periodontal cosmetic surgery. AB - Periodontal plastic procedures are performed to prevent or correct anatomical, developmental, traumatic, or plaque induced defects of the gingiva, alveolar mucosa, or bone. The majority of these procedures are performed in combination with restorative and/or orthodontic therapy with the primary goal of enhancing aesthetics. In this review some of the more prominent techniques currently available to address mucogingival deficiencies including pedicle grafts, free soft tissue grafts, and combination grafts are illustrated. In addition, potential complications associated with periodontal plastic procedures are discussed. PMID- 10833289 TI - Cyclosporin A effects on extra oral connective tissue in heart transplanted patients. AB - Gingival overgrowth (GO) is an adverse side-effect of Cyclosporin A (CsA) therapy. There is wide evidence that fibroblasts are the primary cellular influence on GO and it has been hypothesised that people developing GO have fibroblasts with an abnormal susceptibility to CsA. The purpose of the study was to analyse, on a macroscopic scale, the Cyclosporin effects on soft connective tissues in order to show whether subjects with an abnormal gingival connective reaction to Cyclosporin may have macroscopic signs of a similar reaction in connective tissues other than oral cavity. Twenty-two heart transplanted patients were enrolled in the study. All patients had been on a CsA-based immunosuppression regimen for at least six months (33.2 +/- 20.5). Hyperplastic Index (HI) was considered as the periodontal parameter relating to GO. Macroscopic evidence of the residual cicatrix following thoracotomy (P1) and connective tissue hyperproduction in the subclavian region conditioning the difficulty in probing the vein during myocardial biopsies (P2) were considered as the parameters relating to extraoral cicatricial activity. Both non parametric statistical analysis (Mann Whitney U-test, linear correlation by Spearman) and parametric analysis (Student t-test) were used to investigate the relationship between HI (dependent variable) and P1, P2 (independent variables). Results failed to demonstrate any significant correlation between gingival overgrowth and cicatricial tissue hyperproduction in sites other than the oral cavity. We conclude that the hyperproduction of collagen leading to GO in a certain percentage of subjects treated with CsA is a reaction limited to the oral cavity and not a consequence of a systemic fibroblastic hyperactivity. Our data suggest that the collagen disregulation at gingival level may be a consequence of a local perturbation whose aetiological factors are however still not known. PMID- 10833290 TI - [The effect of two different methods of heel lancing on pain reaction in preterm neonates]. AB - Heel lancing is the most used method to obtain a blood sample in newborn and pre term infants. The effectiveness on pain reduction of a new lancet, Tenderfoot was compared against the traditional Lancetta, randomizing 40 preterm newborns to have the heel lanced with one of the two instruments and observing overall 76 blood sampling episodes. Newborns were observed at baseline, when disinfected, during and at the end of the blood sampling procedure. Statistically different behavioural responses to the pain of the puncture were observed during the blood sampling, in the 'recovery time' after the procedure and in the number of pricks performed. When the Tenderfoot was used, less children cried during the first 5 seconds after the prick. The Tenderfoot is effective in reducing the heel prick pain, allows an easier collection of the blood, a reduction of the time necessary and of the need of squeezing the heel, therefore reducing the bruises and pain related to the squeezing itself. According to our data, its use is recommended for the obtainment of large quantities of blood, and in the severely ill newborns, that need frequent blood monitoring and several pricks. These characteristics render its use debatable because of the costs and of the bleeding for blood exams that require limited amount of blood (blood glucose, blood gas analysis) until its effects on the scarring time and outcome will be documented. PMID- 10833291 TI - [First aid for multiple trauma patients: investigative survey in the Firenze Bologna area]. AB - Overall mortality ascribable to multiple traumas, that in Italy is responsible for about 8,000 death/year, is strictly dependent on the function of the so called Trauma Care System. This study reports on an epidemiological survey conducted in the urban area of Florence along a 23-month period (from Jan 97 to Nov 99), with the aim to identify the typology of traumas and the first aid care delivered to the person until hospital admission. These data were compared to those collected in the urban area of Bologna because the composition of the first aid team is different, being nurses, in Bologna, an integral component of the first aid system. On a total of 118 multiple traumas, 17% was represented by isolated head trauma, while in 72% involvement of other organs was present in addition to the head; 11% of cases were abdominal or thoracic traumas, 1% of lower extremities. In 46% the cause of trauma was a car accident. The complexity of care delivered to the person with trauma was less in the Florence survey, as indicated by the immobilization of patients, performed in only 11% of cases as compared to 47% in Bologna, by the application of the cervical collar, applied in 12% versus 62% of traumas. Although the two samples are not strictly comparable, these data suggest that the presence of nurses in the Trauma Care System can be one of the elements of improvement of the quality of delivered care. PMID- 10833292 TI - [Criteria for the evaluation of environmental hygiene]. AB - The process of rationalizing resources brought the health care institutions and hospitals to let out some services, including the cleaning and sanitation services. In order to write the tender the hospital had to specify both the expected standard of service (i.e. what should be cleaned, how, how many times etc) and the evaluation criteria. The experiences of two community hospitals (Vigevano and Garbagnate) are presented, focusing on the method and criteria they adopted. In both experiences the head nurses were the main responsible for the definition of the criteria, the forms and the process of evaluation. Garbagnate hospital gave priority to outcome criteria (i.e. lack of dust on surfaces, lack of spots on heaters etc.). The 84 criteria identified were divided as follows: 7 criteria for the structure and resources, 6 for the process and 71 for the outcome, with a yes-no-not applicable evaluation. The focus of the other hospital was on process criteria (to be assessed on a 5 points Likert scale, from absent to 100% compliance) and on the knowledge-educational level of the personnel responsible for the cleaning. PMID- 10833293 TI - [Evaluation of the quality and results of cleaning and sanitation services in a Milanese general hospital]. AB - The evaluation of the cleaning and sanitation service involved the head nurses of 7 different wards and 7 health services. The completion of 368 forms allowed to have a clear picture of the performance of the cleaning service. Better results were obtained for the structure, resource and performance criteria than for the outcome criteria. In fact the structure and process criteria were fulfilled in the 75% of the observations whilst the outcome criteria for the ward room were fulfilled in 56% of the observations. The overall agreement between observed performance and adopted standard is satisfactory. The evaluation method identified and the forms used allowed an objective assessment assess the performance of the cleaning services and to the identification of areas for improvement. PMID- 10833295 TI - [The management of emergencies]. PMID- 10833294 TI - [Educational packages: adult education about basic and advanced methods of life support measures. Analysis of some educational systems]. AB - Educational courses are increasingly being offered in Italy to health care workers who want to acquire or improve their basic (BLS) or advanced (ACLS) resuscitation skills. Since 1994 the Italian Resuscitation Council planned and implemented educational BLS and ACLS packages. Their content, methodology and implementation strategies was analysed on the basis of the educational and informational materials offered to the learners and instructors. The duration of the courses varies (never exceeding the three days): the performance of the participants is evaluated and certified. The courses are open to critical care doctors, nurses, technicians and in some cases to the lay people. They are based on the adult learning principles and the learning strategies are both formal and interactive. The course instructors are doctors and nurses specifically trained after a successful result (with optimal performance) of a BLS and ACLS course. At least 50,000 people attended to a BLS course and a lower number (1270) to an ACLS course. To date, the number of recognised BLS instructors is 229 and 79 for the ACLS courses. The recommended costs are around 120 US $ for a BLS course and 400 US $ for an ACLS course. Although the debate on the effectiveness of such a training on the survival of people treated for a cardiac arrest is still ongoing, the learning packages show the feasibility of a multi-professional approach to a systematic training in a critical health area. PMID- 10833296 TI - [Burnout in health professionals: theoretical aspects and prospectives of intervention]. PMID- 10833297 TI - [A silent genocide]. PMID- 10833298 TI - [Beyond type I spinal atrophy]. PMID- 10833299 TI - Temporal appearance of the presynaptic cytomatrix protein bassoon during synaptogenesis. AB - Bassoon is a 420-kDa presynaptic cytomatrix protein potentially involved in the structural organization of neurotransmitter release sites. In this study, we have investigated a possible role for Bassoon in synaptogenesis and in defining synaptic vesicle recycling sites. We find that it is expressed at early stages of neuronal differentiation in which it is selectively sorted into axons. As synaptogenesis begins, Bassoon clusters appear along dendritic profiles simultaneously with synaptotagmin I, sites of synaptic vesicle recycling, and the acquisition of functional excitatory and inhibitory synapses. A role for Bassoon in the assembly of excitatory and inhibitory synapses is supported by the colocalization of Bassoon clusters with clusters of GKAP and AMPA receptors as well as GABA(A) receptors. These data indicate that the recruitment of Bassoon is an early step in the formation of synaptic junctions. PMID- 10833300 TI - Normal phototransduction in Drosophila photoreceptors lacking an InsP(3) receptor gene. AB - The Drosophila light-sensitive channels TRP and TRPL are prototypical members of an ion channel family responsible for a variety of receptor-mediated Ca(2+) influx phenomena, including store-operated calcium influx. While phospholipase Cbeta is essential, downstream events leading to TRP and TRPL activation remain unclear. We investigated the role of the InsP(3) receptor (InsP(3)R) by generating mosaic eyes homozygous for a deficiency of the only known InsP(3)R gene in Drosophila. Absence of gene product was confirmed by RT-PCR, Western analysis, and immunocytochemistry. Mutant photoreceptors underwent late onset retinal degeneration; however, whole-cell recordings from young flies demonstrated that phototransduction was unaffected, quantum bumps, macroscopic responses in the presence and absence of external Ca(2+), light adaptation, and Ca(2+) release from internal stores all being normal. Using the specific TRP channel blocker La(3+) we demonstrated that both TRP and TRPL channel functions were unaffected. These results indicate that InsP(3)R-mediated store depletion does not underlie TRP and TRPL activation in Drosophila photoreceptors. PMID- 10833301 TI - Sequential activation of the 5-HT1(A) serotonin receptor and TrkB induces the serotonergic neuronal phenotype. AB - Serotonin (5-HT) is an important factor controlling survival, differentiation, and plasticity of neurons in serotonergic target regions of the brain and has been implicated in major psychiatric and autonomic disorders. Relatively little is known, however, of factors controlling differentiation and plasticity of developing and adult 5-HT neurons. We show now that 5-HT, the 5-HT1(A) receptor, brain-derived neurotrophic factor (BDNF), and its receptor, trkB, form an auto/paracrine loop for the regulation of the serotonergic phenotype. Serotonin applied to cultures from E14 rat raphe increased numbers of neurons expressing serotonergic markers in a dose-dependent manner. Agonists of the 5-HT1(A) receptor, BP-554 and 8-OH-DPAT, but not agonists of the 5-HT1(B) and 5-HT1(D) receptors, mimicked this effect, while the specific 5-HT1(A) antagonist, WAY 100635, inhibited it. Serotonin also increased BDNF mRNA and protein in embryonic raphe cultures. Induction of serotonergic markers by serotonin was suppressed by a trkB-IgG fusion protein but not by trkC-IgG. Taken together, our data indicate that serotonin acts on 5-HT1(A) autoreceptors, causing up-regulation of BDNF, which activates trkB to promote serotonergic phenotype-specific markers. PMID- 10833302 TI - INP, a novel N-cadherin antagonist targeted to the amino acids that flank the HAV motif. AB - The classical cadherins are homophilic binding molecules that play fundamental roles in several biological processes, including axonal growth and synaptic plasticity. The structures of the amino-terminal homophilic binding domains of N cadherin and E-cadherin have been resolved. However, the mechanisms that govern cadherin binding and specificity remain contentious. In the present study we have used a peptide competition approach to probe for small linear determinants of cadherin binding. We demonstrate that a linear peptide mimetic of a short sequence in ECD1 of N-cadherin (INPISGQ) functions as a highly specific and potent antagonist of N-cadherin function with an IC(50) value of approximately 15 microM. Peptide mimetics of the corresponding motif in chick R-cadherin also inhibited N-cadherin function, albeit with lower efficacy. In contrast, peptide mimetics of the corresponding motif in E- or P-cadherin failed to inhibit N cadherin function. A short cyclic peptide that contained only the INP motif from N-cadherin was also a potent N-cadherin antagonist (IC(50) approximately 15 microM). Analysis of existing crystal structures suggests that the peptides are likely to antagonize N-cadherin function by binding to the region that flanks the HAV motif at the adhesion dimer interface. PMID- 10833303 TI - Neuronal release of vasoactive intestinal peptide is important to astrocytic protection of neurons from glutamate toxicity. AB - Astrocytes regulate clearance of glutamate from the vicinity of neurons. This helps to protect neurons directly from glutamate toxicity. Recent findings have indicated that a complex molecular interaction between neurons and astrocytes that is necessary for this protection occurs. In the present investigation the role of vasoactive intestinal peptide (VIP) in signaling between neurons and astrocytes was investigated. VIP was found to be necessary for the protective effects of astrocytes in a coculture system. VIP in combination with neuronal conditioned medium enhanced glutamate uptake by astrocytes. Also, VIP enhanced the expression of the high-affinity VIP receptor, increased astrocytic release of interleukin-6, and indirectly reduced the toxicity of glutamate in neuronal conditioned astrocyte medium. These results indicate that VIP is essential to the molecular interaction of neurons and astrocytes and is involved in the regulation of the protective effects of astrocytes for neurons. PMID- 10833304 TI - Differential distribution of three Ca(2+)-activated K(+) channel subunits, SK1, SK2, and SK3, in the adult rat central nervous system. AB - Ca(2+)-activated, voltage-independent K(+) channels are present in most neurons and mediate the afterhyperpolarizations (AHPs) following action potentials. They present distinct physiological and pharmacological properties and play an important role in controlling neuronal firing frequency and spike frequency adaptation. We used in situ hybridization to characterize the distribution patterns of the three cloned SK channel subunits (SK1-3), the prime candidates likely to underlie Ca(2+)-dependent AHPs in the central nervous system. We found high levels of expression in regions presenting prominent AHP currents, such as, for example, neocortex and CA1-3 layers of the hippocampus (SK1 and SK2), reticularis thalami (SK1 and SK2), supraoptic nucleus (SK3), and inferior olivary nucleus (SK2 and SK3). Our results reveal the functional role of SK channels with defined subunit compositions in some neurons and open the way to the identification of the molecular determinants of AHP currents in many brain regions. PMID- 10833305 TI - Vulnerability to calcium-induced neurotoxicity in cultured neurons expressing calretinin. AB - Calretinin (CR) is a calcium-binding protein purported to have neuroprotective properties. This study was designed to characterize the types of neurons containing CR in two different primary cultures and to determine which, if any, CR-immunoreactive (CR-ir) neurons are resistant to excitotoxic insults. Calretinin-containing neurons in cortical primary cultures derived from E14 rat embryos were not resistant to either kainic acid or a brief calcium overload induced by the calcium ionophore A23187. Equal proportions of CR-ir and GABAergic cortical neurons were lost after a 24-h exposure to 100 or 500 microM kainic acid. A 3 microM, 3-h exposure to A23187 induced equivalent amounts of cell loss in both the total cell and CR-ir cortical neuron culture populations. Cortical cultures grown for 6-7 days were more vulnerable than 12- to 13-day-old cultures to short-term, low-concentration treatments of A23187. Older cultures, however, were more severely affected when examined 24 h after a 3-h exposure to A23187. Calretinin-immunoreactive neurons derived from the diencephalon were relatively more resistant than cortical neurons to kainic acid at 6-7 days in vitro. In cortical or diencephalic cultures, CR was rarely coexpressed with GABA or calbindin D-28k. No vasoactive intestinal peptide, substance P, or parvalbumin was detected in CR-ir neurons in either culture system. We suggest that the presence of CR alone is not sufficient to spare neurons from a toxic calcium overload. Calretinin may still buffer calcium at low concentrations or be a component in a calcium-based signal transduction system. PMID- 10833306 TI - Cytotoxicity by matrix metalloprotease-1 in organotypic spinal cord and dissociated neuronal cultures. AB - Extracellular matrix (ECM) proteins, including collagens and laminins, are critical to the structure of the neuronal synapse and may also be involved in cell survival. In the present study, we therefore examined the possibility that select ECM degrading proteins might be toxic to organotypic spinal cord and dissociated neuronal cultures. Of those proteins tested, including MMP-1, -7, and -9, we observed that MMP-1 was toxic to spinal cord cultures as determined by release of lactic acid dehydrogenase as well as uptake of propidium iodide. Pretreatment of cell cultures with 50 microM alpha-tocopherol partially reversed these effects. We also observed that MMP-1 was toxic to human neurons grown in dissociated cultures and that increased amounts of MMP-1 were released by astrocytes following their stimulation with IL-1beta. These results suggest that further studies may be warranted to determine whether MMP-1 contributes to neurodegenerative conditions in which activated astrocytes may play a role. PMID- 10833307 TI - Adrenal medullary explants as an efficient tool for pain control: adhesive biomolecular components are involved in graft function ex vivo. AB - Adrenal medullary (AM) tissue transplantation into the central nervous system has been reported as a potential source of opioid peptides and catecholamines, which have analgesic effects useful in the control of chronic pain. Clinical trials, involving allogeneic graft of whole tissue explants into the subarachnoid space of the lumbar spinal cord, have already been reported. The aim of the present study was to determine whether adhesion and function of AM explants were related in some extent and how this relationship could account for improvement of AM tissue in terms of analgesic activity before grafting. Our experiments demonstrated a significant correlation between the adherent state of AM organoids during culture and a sustained secretion of Met-enkephalin and catecholamines by chromaffin cells (CC). These findings suggest that optimal culture condition for AM organoid adhesion can be defined for maintenance of tissue, prior to transplantation. Using immunocytochemistry, flow cytometry, and ELISA assays we showed that different cell adhesion molecules (CAMs) and extracellular matrix ECM proteins were expressed and released by AM cells during culture. Adherent AM organoids expressed increased levels of specific neural CAMs (NCAM and HNK-1 epitope) and integrin chains (beta1, alpha1, alpha2, alpha4, alpha5) and deposited markedly higher levels of fibronectin, but also laminin and collagen IV. Those molecules and probably adhesion processes they control might be involved in the maintenance of the CC-secreting neuroendocrine phenotype through cellular signaling pathways. PMID- 10833308 TI - A quantitative study of the pathological lesions in the neocortex and hippocampus of twelve patients with corticobasal degeneration. AB - The density of ballooned neurons (BN), tau-positive neurons with inclusion bodies (tau+ neurons), and tau-positive plaques (tau+ plaques) was determined in sections of the frontal, parietal, and temporal lobe in 12 patients with corticobasal degeneration (CBD). No significant differences in the mean density of BN and tau+ neurons were observed between neocortical regions. In the hippocampus, the densities of BN were significantly lower than in the neocortex, and densities of tau+ neurons were greater in sectors CA1 and CA2, compared with CA3 and CA4. Tau+ plaques were present in one or more brain regions in six patients. Significantly more BN were recorded in the lower (laminae V/VI) compared with the upper cortex (laminae I/II/III) but tau+ neurons were equally frequent in the upper and lower cortex. No significant correlations were observed between the densities of BN and tau+ neurons, but the densities of BN in the superior temporal gyrus and tau+ plaques in the frontal cortex were positively correlated with age. A principal components analysis (PCA) suggested that differences in the density of tau+ neurons in the frontal and motor cortex were the most important sources of variation between patients. In addition, one patient with a particularly high density of tau+ neurons in the hippocampus appeared to be atypical of the patient group studied. The data support the hypothesis that, although clinically heterogeneous, CBD is a pathologically distinct disorder. PMID- 10833309 TI - Fibroblast growth factor treatment produces differential effects on survival and neurite outgrowth from identified bulbospinal neurons in vitro. AB - The in vivo application of appropriate trophic factors may enhance regeneration of bulbospinal projections after spinal cord injury. Currently, little is known about the sensitivities of specific bulbospinal neuron populations to the many identified trophic factors. We devised novel in vitro assays to study trophic effects on the survival and neurite outgrowth of identified bulbospinal neurons. Carbocyanine dye crystals implanted into the cervical spinal cord of embryonic day (E)5 chick embryos retrogradely labeled developing bulbospinal neurons. On E8, dissociated cultures containing labeled bulbospinal neurons were prepared. Fibroblast growth factor (FGF)-2 (but not FGF-1) promoted the survival of bulbospinal neurons. FGF receptor expression was widespread in the E8 brainstem, but not detected in young bulbospinal neurons, suggesting that nonneuronal cells mediated the FGF-stimulated survival response. Astrocytes synthesize a variety of trophic factors, and astrocyte-conditioned medium (ACM) also promoted the survival of bulbospinal neurons. As might be expected, FGF-2 function blocking antibodies did not suppress ACM-promoted survival, nor did an ELISA detect FGF-2 in ACM. This suggests that nonneuronal cells synthesize other factors in response to exogenous FGF-2 which promote the survival of bulbospinal neurons. Focusing on vestibulospinal neurons, dissociated (survival assay) or explant (neurite outgrowth assay) cultures were prepared. FGF-2 promoted both survival and neurite outgrowth of identified vestibulospinal neurons. Interestingly, FGF-1 promoted neurite outgrowth but not survival; the converse was true of FGF-9. Thus, differential effects of specific growth factors on survival or neurite outgrowth of bulbospinal neurons were distinguished. PMID- 10833310 TI - Reactions of oligodendrocytes to spinal cord injury: cell survival and myelin repair. AB - The aim of this study was to elucidate whether oligodendrocytes die in fiber tracts that are spared by a spinal cord injury but are in close vicinity of inflammatory cells. Adult rat spinal cords were studied histologically 1 day to 2 weeks after a contusion lesion that left the ventral white matter largely intact. Massive oligodendrocyte death occurred in the lesion center, along with the death of neurons, microglia, and astrocytes. Oligodendrocytes, specifically positive for proteolipid protein (PLP) mRNA, were counted in the ventral white matter where axons at the rostral and caudal edges of the lesion were histologically intact. Although these regions contained many macrophages and neutrophils hypothesized to contribute to secondary tissue loss, there was no significant loss of oligodendrocytes. In the ventral funiculus, 3 and 6 mm rostral and caudal to the lesion, oligodendrocyte numbers were also unchanged, in spite of the presence of many activated microglial cells. From day 7 on, oligodendrocytes in close vicinity to the lesion increased their expression of PLP mRNA. We conclude that, at least within the first 2 weeks after a spinal cord contusion lesion, there is no major devastating influence of inflammatory cells or their mediators on oligodendrocytes. When death occurs, it may be due to mechanical trauma, ischemia, or excitotoxicity within the lesion or it may occur as a result of axonal degeneration. PMID- 10833311 TI - Abnormal glutamate transport function in mutant amyloid precursor protein transgenic mice. AB - Recent studies have shown that amyloid precursor protein (APP), which plays a central role in Alzheimer's disease (AD), protects against excitotoxic neuronal injuries by regulating the function of the glial glutamate transporters. The mechanisms underlying these effects and their relationship to the neurodegenerative process in AD are under intense scrutiny. In this context, the main objective of the present study was to determine if overexpression of mutant human APP in transgenic mouse brains results in altered functioning of the excitatory amino acid transporters (EAATs). Transgenic mice expressing the 695 amino acid form of the human APP from the Thy-1 promoter showed a significant decrease in B(max) and K(D) for aspartate uptake when compared to nontransgenic controls. This decrease in glutamate transporter activity was associated with decreased protein expression of glial specific glutamate transporters, EAAT1 and 2, but did not affect mRNA levels. These results suggest that expression of mutant forms of APP disturbs astroglial transport of excitatory amino acids at the posttranscriptional level leading, in turn, to increased susceptibility to glutamate toxicity. PMID- 10833312 TI - Interleukin-1 promotes expression and phosphorylation of neurofilament and tau proteins in vivo. AB - Slow-release, IL-1-impregnated pellets implanted in rat cerebral cortex to simulate chronic overexpression of IL-1 significantly increased relative tissue levels of tau mRNA and of tau immunoreactivity in neuronal cell bodies and in swollen dystrophic neurites that also overexpressed phosphorylated and nonphosphorylated neurofilament epitopes. In addition, rats with IL-1-impregnated pellets, but not control rats or those with sham pellets, showed focal immunoreactivity for hyperphosphorylated tau epitopes present in paired helical filaments. Our results are consistent with an important driving role for IL-1 in the pathogenesis of Alzheimer-type neuronal and neuritic changes. PMID- 10833313 TI - Mice overexpressing extracellular superoxide dismutase have increased resistance to global cerebral ischemia. AB - Transgenic mice, which exhibit a fivefold increase in brain parenchymal extracellular superoxide dismutase (EC-SOD) activity, were used to investigate the role of EC-SOD in global ischemic brain injury. Halothane-anesthetized normothermic wild-type (n = 22) and transgenic (n = 20) mice underwent 10 min of near-complete forebrain ischemia induced by bilateral carotid artery occlusion and systemic hypotension (mean arterial pressure = 30 mm Hg). After 3 days of recovery, the brains were histologically examined. Other mice underwent autoradiographic determination of regional CBF 10 min prior to, during, and 30 min after forebrain ischemia. Histologic injury in the cortex and caudoputamen was minimal in both groups. The percentage of dead hippocampal CA1 neurons was reduced in the EC-SOD transgenic group (wild type = 44 +/- 28%; EC-SOD transgenic = 23 +/- 21%, mean +/- SD, P = 0.015). CBF was similar between groups prior to ischemia. The intraischemic blood flow was severely reduced in forebrain structures and was similar between groups. Blood flow at 30 min postischemia had recovered to 50-60% of baseline values in both groups. These results indicate that EC-SOD can play an important role in defining the magnitude of selective neuronal necrosis resulting from near-complete forebrain ischemia. This implicates involvement of extracellular superoxide anions in the pathologic response to global cerebral ischemia. PMID- 10833314 TI - Postischemic spontaneous hyperthermia and its effects in middle cerebral artery occlusion in the rat. AB - This study examined the time course and effects of postischemic spontaneous hyperthermia after transient and permanent focal ischemia. Rats underwent a 90 min, 120-min, or permanent middle cerebral artery occlusion (MCAO). Body temperatures started rising 15-20 min after MCAO and reached 39-40.5 degrees C during the first hour. Sustained hyperthermia was observed during the rest of the first 24 h. In another experiment, rats were subjected to the same interventions, but a normothermic body temperature was maintained. Spontaneous hyperthermia significantly increased the infarct volumes measured 48 h after MCAO in all groups. Reperfusion 2 h after the onset of ischemia was not beneficial in the hyperthermic animals in contrast to the normothermic group. We also examined the effect of spontaneous hyperthermia on the temporal progression of infarcted and penumbral areas 4, 12, or 48 h after MCAO. During spontaneous hyperthermia, penumbral areas became infarcted areas more rapidly, which was most expressed at 4 h. These findings demonstrate that severe spontaneous hyperthermia can occur in rats after MCAO and that it not only increases the infarct volumes in both transient and permanent ischemia, but also accelerates the incorporation of penumbral areas into necrotic areas, which significantly decreases the window of opportunity for therapeutic interventions. PMID- 10833315 TI - Enlargement of the receptive field size to low intensity mechanical stimulation in the rat spinal nerve ligation model of neuropathy. AB - One characteristic of plasticity after peripheral tissue or nerve damage is receptive field reorganization, and enlargement of receptive field size has been suggested to occur in certain models of neuropathic pain. The aim of the present study was to explore whether enlargement of neuronal receptive fields could contribute to the mechanical allodynia found on the ipsilateral paw in the spinal nerve ligation model of neuropathy. After ligation of L(5)-L(6) spinal nerves, all rats developed behavioral signs of mechanical allodynia, while the sham operated control group displayed no such changes. The characteristics of the evoked responses of the neurones recorded in the dorsal horn of the rats were similar between the spinal nerve ligation, the sham operated control group, and the nonoperated control group, except for spontaneous activity, which was significantly increased in the spinal nerve ligation group. The mean size of the receptive field on the ipsilateral hindpaw, mapped using low-intensity stimulation with 9-g von Frey hair, was significantly increased in the spinal nerve ligation group, as compared to the sham-operated group. No significant difference was seen with 15- or 75-g von Frey hairs. The distribution of the receptive fields over the plantar surface of the paw was similar between the study groups. The enlargement of receptive field for non-noxious touch could be an indication of central sensitization in this model. PMID- 10833316 TI - Altered ionic conductances in axons of transgenic mouse expressing the human neurofilament heavy gene: A mouse model of amyotrophic lateral sclerosis. AB - Neurofilaments (NFs; made by copolymerization of three intermediate filament proteins NF-L, NF-M, and NF-H, for light, medium, and heavy) constitute the most abundant cytoskeletal structure in large myelinated axons. The presence of aberrant NF accumulation has been associated with neurodegenerative diseases (such as ALS). The possible causal role of NF in neurodegeneration has been supported by studies on recently available transgenic mice in which expression of human NF-H (hNF-H +/+) leads to overt neuropathy. We have examined electrophysiological properties of myelinated axons in hNF-H +/+ mice using intraaxonal microelectrode recording from isolated sciatic and tibial nerves. Transgenic mice showed several deficits in physiological properties of low threshold myelinated fibers: conduction velocity and resting membrane potential were significantly decreased (20 +/- 1.6 vs 40 +/- 2 m/s; -71.3 +/- 0.9 vs -75.5 +/- 0.5 m/s; mean +/- SE; n = 25; 22 degrees C). While the amplitude of action potentials was of comparable size (82 +/- 5 vs 86 +/- 3 mV) duration of action potential (at half-amplitude, AP/2) in hNF-H +/+ was significantly prolonged (0.82 +/- 0.02 vs 0.65 +/- 0.02 ms). Voltage-current properties of axonal membrane indicate a significant decrease in inward and outward rectification. Occasionally, impaled axons of hNF-H +/+ showed membrane oscillations and repetitive activity (reminiscent of fasciculations) never observed in normal animals. These results are compatible with an imbalance between ion conductances in axons from transgenic animals (an increase in Na(+) and a decrease in K(+) conductances), in agreement with recent suggestion based on clinical studies on ALS patients (H. Bostock et al., 1995, Brain 118, 217-225). One may hypothesize that these changes could contribute to neurodegenerative processes (i.e., via an increase in [Na(+)](i)), as well as clinical symptoms (fasciculations) observed in patients with degenerative motor neuron diseases. PMID- 10833317 TI - Spinal cord microstimulation generates functional limb movements in chronically implanted cats. AB - Spinal cord injuries disrupt the communication between the brain and peripheral nerves, but leave motoneurons and networks of interneurons below the level of the lesion intact. It is therefore possible to restore some function following injury by providing an artificial stimulus to the surviving neurons below the level of the lesion. We report here on a novel approach for generating functional movements by electrically stimulating the spinal cord through chronically implanted ultrafine, hair-like electrodes. Six to 12 microwires were implanted in the lumbar enlargement of intact cats for 6 months. Twice a week, trains of stimuli were delivered through each microwire and the evoked electromyographic and torque responses were recorded. Strong coordinated hindlimb movements were obtained by stimulating through individual electrodes. The joint torques elicited were capable of supporting the animals' hindquarters. The responses were stable over time and the contractions caused no apparent discomfort to the animals. No obvious motor deficits were seen throughout the 6-month duration of implantation. The results demonstrate that microwires implanted in the spinal cord remain stably in place and stimulation through these electrodes produces strong, controllable movements. This provides a promising basis for the use of spinal cord neuroprostheses in restoring mobility following spinal cord injury. PMID- 10833318 TI - Quinolinic acid released from polymeric brain implants causes behavioral and neuroanatomical alterations in a rodent model of Huntington's disease. AB - Quinolinic acid (QA) is an N-methyl-d-aspartate agonist that has been shown to produce neurotoxic effects that mimic certain neurodegenerative diseases when administered to laboratory animals. Intrastriatal injections of QA in rats have been used extensively to produce some of the neuropathological and behavioral deficits that are analogous to Huntington's disease (HD). However, acute intrastriatal injections of QA produce symptoms that are not analogous to the progressive nature of HD. Thus far, models using chronic administration of QA that produce HD-like behavioral and neuroanatomical changes have necessitated the use of a relatively bulky and fragile microdialytic pump apparatus. The present study tested an alternative way of chronically administering QA. Specifically, this study tested whether gradual release of QA from ethylene vinylacetate (EVA) polymers could produce symptoms analogous to HD. Rats received either no implants or bilateral intrastriatal implants of polymers with or without QA. Subsequent tests for spontaneous motor activity (SMA), grip strength, balance, and learning ability in a radial-arm-water-maze task revealed QA-induced impairments in balance and learning ability, but did not affect grip strength or SMA. Histological analysis revealed QA-induced enlargement of lateral ventricles, striatal atrophy, and striatal neuronal loss, with relative sparing of NADPH diaphorase-positive neurons. These results suggest that QA released from polymers can produce behavioral and neuropathological profiles analogous to early stages of HD and that EVA polymers offer a useful means of chronically delivering QA in rodent models of neurodegeneration. PMID- 10833319 TI - Decreased hippocampal metabolic activity in Alzheimer patients is not reflected in the immunoreactivity of cytochrome oxidase subunits. AB - In the present study we have compared histochemically determined cytochrome oxidase activity with the levels of immunocytochemically stained cytochrome oxidase subunits (CO II and CO IV) and ATP synthase in the human hippocampus in relation with Alzheimer's disease. Cytochrome oxidase activity was significantly reduced in all hippocampal areas of Alzheimer patients. The protein levels of subunits II and IV were not different between control subjects and Alzheimer patients. Additionally, it was observed that the active cytochrome oxidase is evenly distributed over both cell bodies and neuropil, while a relatively large pool of inactive enzyme or precursors is limited to the neuronal somata. Further, in Alzheimer patients the CO IV immunoreactivity decreased with age, whereas in control subjects it increased with age. Our results suggest that the assembly of cytochrome oxidase or the processing of its subunits may be impaired. PMID- 10833320 TI - Apolipoprotein E epsilon4 allele has no effect on age at onset or duration of disease in cases of frontotemporal dementia with pick- or microvacuolar-type histology. AB - Frontotemporal dementia (FTD) is the second most common cause of presenile dementia. Here we have investigated the frequency of the epsilon4 allele of the Apolipoprotein (APOE) gene in FTD and in other non-Alzheimer forms of dementia related to FTD such as Motor Neurone disease dementia, semantic dementia, progressive aphasia, progressive supranuclear palsy, and corticobasal degeneration. In none of these diagnostic groups did we find a significant increase in the APOE epsilon4 allelic frequency, compared to population values. Neither did we observe any affects of the epsilon4 allele upon age at onset or duration of disease. We conclude therefore that polymorphic variations in the APOE gene do not modulate either the occurrence or progression of these non Alzheimer forms of dementia. PMID- 10833322 TI - Role of the forebrain in bladder overactivity following cerebral infarction in the rat. AB - This study was undertaken to investigate the contribution of the forebrain to bladder overactivity induced by cerebral infarction (CI). CI was induced by left middle cerebral artery (MCA) occlusion in female SD rat. Two and a half hours after CI or a sham operation (SO) decerebration was performed in some animals to eliminate forebrain influences on voiding function. Then bladder activity was monitored during continuous infusion cystometrograms in awake rats for 2.5 h. The effects of cumulative intravenous doses of MK-801 (0.1-1.4 mg/kg), an NMDA (N methyl-D-aspartate) glutamatergic receptor antagonist, or sulpiride (0.1-41.1 mg/kg), D(2) selective dopaminergic receptor antagonists were studied over a 1.5 h period beginning 5 h after MCA occlusion. Bladder capacity was reduced by 57.5% after CI. In CI rats decerebration increased bladder capacity by 62.5% of predecerebration capacity. In SO rats bladder capacity was reduced by 25% after decerebration. MK-801 (0.4 and 1.4 mg/kg) increased bladder capacity in CI and CI decerebrate rats, but did not change bladder capacity in SO-decerebrate rats. MK 801 decreased (60.7%) bladder capacity in SO-nondecerebrate rats. Sulpiride (11.1 and 41.1 mg/kg) significantly increased bladder capacity in CI, CI-decerebrate, and SO-decerebrate rats, but had no effect in SO-nondecerebrate rats. These results indicate that CI-induced decrease in bladder capacity is mediated by two mechanisms: (1) upregulation of an excitatory pathway from the forebrain, an effect blocked by decerebration and (2) downregulation of a tonic inhibitory pathway from the forebrain. The latter effect which can be induced by decerebration as well as CI unmasks a D(2) dopaminergic excitatory mechanism. An NMDA excitatory mechanism also contributes to the bladder overactivity after CI, but not after decerebration. PMID- 10833321 TI - Calreticulin binding and other biological activities of survival peptide Y-P30 including effects of systemic treatment of rats. AB - Neuron survival-promoting peptide Y-P30, purified from oxidatively stressed neural cell lines, inhibits the appearance of microglia and rescues neurons 1 week after direct application to lesions of the rat cerebral cortex (7). Y-P30 affinity matrices treated with solubilized membranes from a variety of cell lines including human neuroblastoma SY5Y, mouse hippocampal cells HN 33.1, and human promonocytes HL-60, as well as with cerebral cortex tissue from both humans and rats, showed highly specific binding to calreticulin, a ubiquitous calcium binding protein that may be critical for integrin function. Treatment of cultures with 0.1 nM Y-P30 stabilized all these cell types whether differentiated or not, while 1 microM peptide also inhibited the morphological differentiation of the HL 60 cells into macrophages. Western analysis of the medium of SY5Y cell cultures suggested Y-P30-stimulated release of calreticulin, a result consistent with its other biological activities. Likewise, single dose systemic application of Y-P30 in unoperated rats and in rats with cerebral cortex lesions produced significant reductions in cerebral cortex membrane-associated calreticulin. Both direct and intravenous treatment with peptide also reduced cortical neuron atrophy 4 days after these lesions but only direct application consistently inhibited the appearance of ED-1(+) monocyte derivatives. We suggest that in vitro and in vivo mechanisms of Y-P30 effects are similar and involve the targeting of calreticulin. The results also suggest that some of these activities are apparent in the cerebral cortex after systemic application of this peptide. PMID- 10833323 TI - Synergistic combinatorial stroke therapy: A quantal bioassay of a GABA agonist and a glutamate antagonist. AB - We sought to prolong the window for stroke treatment using synergistic combinatorial therapy. We used the intraluminal filament occlusion model in rats to cause focal cerebral ischemia and a quantal bioassay to measure efficacy. The GABA agonist muscimol and the glutamate antagonist MK-801 were used alone and in combination at various times after ischemia onset. At progressively longer treatment delay intervals (30, 60, 75, 120, 240, and 360 min), higher doses of the single drugs were required to achieve neuroprotection. In contrast, the combination 1.0 mg/kg muscimol plus 0.5 mg/kg MK-801 was effective at all delay intervals studied except the longest (P < 0.05 at each time). After 240 min from ischemia onset, the combination was more effective than either single agent (P < 0.05 for each drug dose), suggesting synergism. The neuroprotective effect could not be demonstrated using morphometry. The treatment effects were probably not due to hypothermia because brain temperatures recorded in awake, unregulated subjects remained normo- or slightly hyperthermic following all treatments. Awake subjects kept on a heating pad exhibited mild brain hyperthermia. The combination caused a drop and MK-801 caused a significant increase in mean arterial blood pressure (main effects F(5,172) = 29, P < 0.0001). The combination of a GABA agonist and glutamate antagonist appears to possess synergistic neuroprotective effects when treatment is delayed up to 240 min following the onset of cerebral ischemia. Temperature regulation causes hyperthermia in awake subjects. The quantal bioassay is one method suitable for studies of synergistic stroke therapy. PMID- 10833324 TI - A behavioral test paradigm to measure the aversive quality of inflammatory and neuropathic pain in rats. AB - The present experiment assessed the aversive quality of neuropathic and inflammatory pain in rats. Compared to sham-treated animals, L5 ligated (neuropathic) and complete Freund's adjuvant (inflammatory)-treated animals displayed an initial period of escape followed by avoidance of a preferred location of the test chamber that was associated with mechanical stimulation of the hyperalgesic paw. The onset of the avoidance behavior occurred during the first 10-15 min of behavioral testing and was maximal at 30 min. It is concluded that animals find mechanical stimulation of the hyperalgesic paw aversive and that this behavioral test paradigm is an additional method that may be used to assess nociception in rat neuropathic and inflammatory models. PMID- 10833325 TI - On neurodegenerative diseases, models, and treatment strategies: lessons learned and lessons forgotten a generation following the cholinergic hypothesis. AB - Life's Journey If life is indeed a journey, then poetry must be the map that reveals all its topographic possibilitiesellipsis while science is the compass that keeps us from getting lost. -R. T. Bartus, Simple Words for Complex Lives, (c) 1998 In the nearly 20 years since the cholinergic hypothesis was initially formulated, significant progress has been achieved. Initial palliative treatments for Alzheimer's disease (AD) have proven beneficial and have gained FDA approval, the use of animal models for studying AD and other neurodegenerative diseases has achieved wider acceptance, and important insight into the potential causes and pathogenic variables associated with various neurodegenerative diseases continues to increase. This paper reviews the current status of the cholinergic hypothesis in the context of continuing efforts to improve upon existing treatments for AD and explores the role that animal models might continue to play. Using the benefit of hindsight, particular emphasis is placed on an analysis of the approaches, strategies, and assumptions regarding animal models that proved useful in developing the initial treatments and those that did not. Additionally, contemporary issues of AD are discussed within the context of the cholinergic hypothesis, with particular attention given to how they may impact the further refinement of animal models, and the development of even more effective treatments. Finally, arguments are presented that, despite the deserved enthusiasm and optimism for identifying means of halting the pathogenesis of AD, a clear need for more effective palliative treatments will continue, long after successful pathogenic treatments are available. This review, therefore, focuses on issues and experiences intended to: (a) facilitate further development and use of animal models for AD and other neurodegenerative diseases, and (b) accelerate the identification of newer, even more effective treatments. PMID- 10833326 TI - Krabbe disease: genetic aspects and progress toward therapy. AB - Krabbe disease or globoid cell leukodystrophy is a disorder involving the white matter of the peripheral and central nervous systems. Mutations in the gene for the lysosomal enzyme galactocerebrosidase (GALC) result in low enzymatic activity and decreased ability to degrade galactolipids found almost exclusively in myelin. The pathological changes observed, including the presence of globoid cells and decreased myelin, appear to result from the toxic nature of psychosine and accumulation of galactosylceramide that cannot be degraded due to the GALC deficiency. Over 60 mutations have been identified in this gene. The great majority are disease-causing; however, a few are considered polymorphisms. While most patients present with symptoms within the first 6 months of life, others present later in life including adulthood. Even patients with the same genotype can have very different clinical presentations and course. The reason for this is not known. Treatment at this time is limited to hematopoietic stem cell transplantation that appears to slow the progression of the disease and improve the magnetic resonance images. Studies using stem cells and viral vectors to transduce transplantable cells are under way in model systems. In culture, oligodendrocytes from the twitcher mouse model can assume a normal appearance after differentiation if GALC activity is provided via viral transduction or uptake from donor cells. Therefore continued myelination and/or remyelination in patients will require supplying GALC activity by transplanted cells or viral vectors to still functional endogenous oligodendrocytes or transplantation of normal oligodendrocytes or stem cells that can differentiate into oligodendrocytes. Using the animal models these options can be explored. PMID- 10833327 TI - Positional cloning utilizing genomic DNA microarrays: the Niemann-Pick type C gene as a model system. AB - A major obstacle in positional cloning is identifying the specific mutated gene from within a large physical contig. Here we describe the application of DNA microarray technology to a defined genomic region (physical map) to identify: (i) exons without a priori sequence data and (ii) the disease gene based on differential gene expression in a recessive disorder. The feasibility was tested using resources from the positional cloning of the Neimann-Pick Type C (NP-C) disease gene, NPC1. To identify NPC1 exons and optimize the technology, an array was generated from genomic fragments of the 110-kb bacterial artificial chromosome, 108N2, which encodes NPC1. First, as a test case for blindly identifying exons, fluorescently labeled NPC1 cDNA identified 108N2 fragments that contained NPC1 exons, many of which also contained intronic sequences and could be used to determine part of the NPC1 genomic structure. Second, to demonstrate that the NPC1 disease gene could be identified based upon differential gene expression, subarrays of 108N2 fragments were hybridized with fluorescently labeled cDNA probes generated from total RNA from hamster cell lines differentially expressing NPC1. A probe derived from the NP-C cell line CT60 did not detect NPC1 exons or other genomic fragments from 108N2. In contrast, several NPC1 exons were detected by a probe generated from the non-NP-C cell line 911D5A13, which was derived from CT60, and expressed NPC1 as a consequence of stable transduction with a YAC that contains NPC1 and encompasses 108N2. Thus, the array technology identified NPC1 as a candidate gene based on a physical contig and differential NPC1 expression between NP-C and non-NP-C cells. This technique should facilitate gene identification when a physical contig exists for a region of interest and mutations result in changes in the mRNA level of the disease gene or portions thereof. PMID- 10833328 TI - Insulin increases leptin mRNA expression in abdominal subcutaneous adipose tissue in humans. AB - Insulin regulates expression and production of leptin in rodents but whether this is also true in humans remains unclear. To test the effects of acute hyperinsulinemia on expression of leptin mRNA in humans, percutaneous needle biopsies of abdominal subcutaneous adipose tissue were performed at baseline and immediately following a 200-min two-step hyperinsulinemic-euglycemic glucose clamp in 16 Pima Indians (8M/8F). Leptin mRNA was quantified by reverse transcription, PCR amplification and expressed relative to actin mRNA. Leptin mRNA levels were higher in women than men (25.6 +/- 1.7 v 16.9 +/- 2.1 relative units, P = 0.003) at baseline. Baseline levels were directly related to percentage body fat (r = 0.54, P = 0. 03) and fasting plasma glucose concentrations (r = 0.57, P = 0.02) and were negatively correlated to glucose disposal at physiologic insulin concentrations (750 +/- 40 pmol/L) during the clamp (r = -0. 51, P = 0.04). Acute hyperinsulinemia (final insulin concentration 11560 +/- 950 pmol/L) increased leptin mRNA levels in 13 of 16 individuals an average of 13% (21.3 +/- 1.7 to 24.2 +/- 1.2 relative units, P = 0.01). Changes in leptin mRNA were directly related to glucose disposal rates during physiologic hyperinsulinemia (r = 0.54, P < 0.04). These results suggest that the expression of leptin mRNA is regulated by insulin in humans, as it is in rodents. PMID- 10833329 TI - Altered folate metabolism and disposition in mothers affected by a spina bifida pregnancy: influence of 677c --> t methylenetetrahydrofolate reductase and 2756a -> g methionine synthase genotypes. AB - Periconceptional folate prevents spina bifida although the mechanisms involved are unclear. We present the genotype frequency for the 677 ct methylenetetrahydrofolate reductase (MTHFR) and 2756ag methionine synthase (MetSyn) polymorphisms. Calculated odds ratios (OR) show that neither the homozygous recessive genotype, carriage of the mutant allele, nor frequency of the mutant allele represent significantly increased risk for neural tube defect (NTD). This is true for both polymorphisms. Simultaneous carriage of t and g alleles is also not a significantly increased risk for NTD. OR and 95% CI for carriage of (i) t allele, (ii) g allele, and (iii) simultaneous carriage of t and g alleles in NTD are 0.89 (0.28-2.82), 0.97 (0.28-3.30), and 0.61 (0.11-3.52), respectively. OR and 95% CI for frequency of t and g alleles are 0.94 (0.42-2.13) and 0.88 (0. 29-2.67), respectively. Unlike some previous studies, we could not detect a significantly increased risk for NTD conferred by the 677ct MTHFR tt genotype; OR 0.98 (0.19-6.49). Differences were found to exist in the circulating whole blood folate profile: total formyl-H(4)PteGlu was significantly higher than total 5-methyl-H(4)PteGlu in control (P = 0.036) but not NTD blood. When broken down into the various 677 ct MTHFR and 2756ag MetSyn genotypes, carriage of the 677ct MTHFR allele appears to affect formyl-H(4)PteGlu metabolism in non-NTD mothers. In addition, NTD mothers exhibited noticeably lower formyl-H(4)PteGlu levels compared to controls; these effects, however, were not significant. 2756ag MetSyn is similarly associated with an altered formyl-H(4)PteGlu disposition. The ag genotype had significantly more formyl-H(4)PteGlu relative to 5-methyl H(4)PteGlu than wildtype 2756ag MetSyn (P = 0.024). This heterozygous increase in the relative formyl-H(4)PteGlu level holds true for controls only; no such relationship occurred in NTD samples. Folyl hexaglutamates are the active cellular coenzyme forms. We showed that where 5-methyl-H(4)PteGlu(6) predominates, Hcy levels are highest. As the relative abundance of formyl H(4)PteGlu(6) increased, so Hcy decreased, presumably due to increased Hcy remethylation, a process in which 5-methyl-H(4)PteGlu(6) is demethylated and downstream folates like formyl-H(4)PteGlu(6) are produced. The negative linear association between the hexaglutamate ratio (formyl-H(4)PteGlu(6)/5-methyl H(4)PteGlu(6)) and Hcy is significant for control (r = -0.64, P = 0.003) but not NTD samples. This effect, centering on Hcy remethylation, is supported by a statistically elevated formyl-H(4)PteGlu(6) to 5-methyl-H(4)PteGlu(6) level in controls relative to NTDs (P = 0.047). The overall (polymorphism independent) effect of exogenous 5,10-methenyl-H(4)PteGlu(1) substrate on the cellular folate profile was to preferentially increase formyl-H(4)PteGlu, while exogenous 5 methyl-H(4)PteGlu(1) substrate dramatically increased metabolic production of 5, 10-methylene-H(4)PteGlu. The following differences were observed between NTD and control samples: (i) a reduced expansion of the formyl-H(4)PteGlu(6) pool in NTD with exogenous 5, 10-methenyl-H(4)PteGlu(1) (P = 0.0005 for control expansion, NS for NTD increase); (ii) a reduced initial expansion of the 5, 10-methylene H(4)PteGlu pool in NTD following treatment with exogenous 5-methyl-H(4)PteGlu(1) substrate (difference between subject groups; P = 0.031). In addition, taking polymorphisms into account, lysate from NTD-MTHFR wildtypes utilized less exogenous 5-methyl-H(4)PteGlu(1) substrate than control-MTHFR wildtypes in the short (P = 0.011) and long term (P = 0.036). Commensurate with this latter effect, the initial production of 5,10-methylene-H(4)PteGlu due to exogenous 5 methyl-H(4)PteGlu(1) substrate was significantly reduced in the NTD-MTHFR wildtype (P = 0.037). These two MTHFR wildtype effects imply that the 677 ct polymorphism is not the only mutation affecting folate metabolism in NTD mothers. (ABSTRACT TRUNCATED) PMID- 10833330 TI - Investigation of folate pathway gene polymorphisms and the incidence of neural tube defects in a Texas hispanic population. AB - Neural tube defects (NTDs) are multifactorial in their etiology, having both genetic and environmental factors contributing to their development. Recent evidence demonstrates that periconceptional supplementation of the maternal diet with a multivitamin containing folic acid significantly reduces the occurrence and recurrence risk for having a pregnancy complicated by NTDs. Unfortunately, the mechanism underlying the beneficial effects of folic acid remains unknown. NTD surveillance data from the Texas-Mexico border show that the high NTD rate (28/10,000 live births) noted during the 1990-1991 Cameron county NTD cluster was superimposed on a background Cameron county NTD rate (16/10,000 live births) which is considerably higher than that generally noted in the United States (8 10/10,000 live births). These data suggest that genetic factors as well as transient environmental factors may contribute to the etiology of the NTDs. Furthermore, clinical and experimental evidence imply that allelic forms of genes involved with folate metabolism and/or transport may explain some of the observed variation in the NTD rates found across different populations. Two folate pathway genes were selected for evaluation in this study. The loci investigated included two known alleles of the 5, 10-methylenetetrahydrofolate reductase (MTHFR) gene, as well as the promoter region of the folate receptor-alpha (FR-alpha) gene. Odds ratios (ORs) for the C677T polymorphism in the MTHFR gene were 1.8 (CI 0.47-6.8) for heterozygosity and 1.8 (CI 0.35-9.4) for homozygosity for the mutant 677T allele, relative to wildtype homozygotes. The odds ratio for the heterozygosity for the A1298C polymorphism in the same gene was 1.1 (CI 0.09-14). No individuals homozygous for the 1298C allele were observed. The OR for heterozygosity of FR alpha gene polymorphisms detected at nucleotide 762 and at nucleotides 610/631 was 1.4 and 0.7, respectively. Neither of the FR-alpha polymorphisms was observed in the homozygous condition. No statistically significant associations were observed for any of the polymorphisms examined, as the 95% confidence intervals for all of the ORs included one. However, the frequency of the MTHFR 677T allele in the largely Hispanic control group from Texas was significantly different from other populations (P < 0.005), and among the highest reported for any control populations examined. PMID- 10833331 TI - Polymorphisms in the CBS gene associated with decreased risk of coronary artery disease and increased responsiveness to total homocysteine lowering by folic acid. AB - Elevated total plasma homocysteine (tHcy) is an established risk factor for the development of vascular disease and neural tube defects. Total homocysteine levels can be lowered by folic acid supplements but individual response is highly variable. In this case-control study, involving 142 coronary artery disease (CAD) patients and 102 controls, we have typed six genetic polymorphisms in three homocysteine metabolizing genes and examined their relationship to the incidence of CAD, tHcy levels, and lowering of tHcy levels in response to folic acid supplementation. We found that two single nucleotide polymorphisms in the cystathionine beta synthase (CBS) gene, 699C --> T and 1080T --> C, are associated with decreased risk of CAD and increased responsiveness to the tHcy lowering effects of folic acid. Individuals homozygous for 699T were significantly underrepresented in CAD patients as compared to controls (4.9% vs 17.3%, P = 0.0015), as were individuals homozygous for the 1080C (29.6% vs 44.2%, P = 0.018). Additionally, 699T and 1080C homozygous individuals were the most responsive to folate supplementation. 699T homozygotes lowered tHcy levels 13.6% on average, compared to 4.8% lowering in 699C homozygotes (P = 0.009), while 1080C homozygotes lowered 12.9% compared to just 2.7% for 1080T homozygotes (P = 0.005). The two polymorphisms in CBS are third codon changes and would not be predicted to affect the underlying protein. However, there is strong linkage disequilibrium between these two positions, suggesting that they may also be linked to other as yet unidentified polymorphisms within the CBS gene. These observations suggest that specific CBS alleles are a risk factor for the development of vascular disease and that genetic information could be predictive of individual response to folic acid supplementation. PMID- 10833332 TI - Inositol phosphoglycans and the regulation of the secretion of leptin: in vitro effects on leptin release from adipocytes and the relationship to obesity. AB - The ratio of two families of inositol phosphoglycans (IPG-A:IPG-P), insulin second messengers, is raised in non-insulin-dependent diabetes mellitus (NIDDM) and obesity. It is shown here that IPG A type inhibits leptin release from adipocytes, contrasting with the action of insulin (stimulation) and IPG P type (no effect). The significance of inhibitory effects of IPG A type on leptin release is important in relation to obesity and NIDDM in view of the action of leptin in promoting Lep expression and fat oxidation in muscle, in addition to its effects on satiety. Energy conservation and oxidation via interorgan regulation by leptin could be compromised by a rise in the IPG-A:IPG-P ratio. PMID- 10833333 TI - The tissue-specific, alternatively spliced single ATG exon of the type 3 voltage dependent anion channel gene does not create a truncated protein isoform in vivo. AB - Voltage-dependent anion channels (VDACs) are small, integral membrane proteins that traverse the outer mitochondrial membrane and conduct ATP and other small metabolites. They are known to bind several kinases of intermediary metabolism in a tissue-specific fashion, have been found in close association with the adenine nucleotide translocator of the inner mitochondrial membrane, and are hypothesized to form part of the mitochondrial permeability transition pore, which results in the release of cytochrome c at the onset of apoptotic cell death. VDACs are found throughout all strata of eukaryotic evolution and exhibit biophysical characteristics that are well conserved from yeast to mammals. The mammalian VDAC gene family consists of three isoforms, each of which shares approximately 70% sequence identity with the other two family members. Recently, we reported that a single codon (ATG) exon is alternatively spliced into the transcript of the type 3 voltage-dependent anion channel (VDAC3) in a tissue-specific fashion. This unusual splicing event was shown to be conserved between mouse and human, and we theorized that the spliced exon could lead to the creation of an alternative translational initiation site. Here we report that a highly specific polyclonal VDAC3 antibody was unable to detect the truncated protein isoform indicative of this putative downstream start site. From these in vivo studies, we conclude that the alternatively spliced exon results in the insertion of a single methionine residue at amino acid position 39 of the mature VDAC3 protein. Additionally, we have used a cross-species genomic sequence comparison to identify conserved regions that may be involved in the regulation of small exon splicing. PMID- 10833334 TI - Molecular cloning and expression profile of rat myocilin. AB - Myocilin is known to be associated with the pathogenesis of juvenile-onset primary open angle glaucoma. The tissue distribution of myocilin transcripts has been analyzed in both humans and mice, and a high level of expression in the retina and skeletal muscle has been reported. The functions of myocilin in these tissues are unknown. We isolated rat myocilin cDNA and examined the expression pattern of myocilin, including its expression in endocrine organs, using Northern blot analysis. The rat myocilin cDNA sequence has two in-frame initiation codons, the upstream and downstream ATGs corresponding to the initiation codon of human and murine myocilin, respectively. It is most likely that the first ATG is a translational initiation codon, since 8 of 13 amino acid residues deduced from the rat cDNA sequence between the first and the second ATGs are the same as those in human myocilin. The open reading frame encodes 502 amino acids. Rat myocilin also has both a myosin-like domain and an olfactmedin-like domain, which have been identified in human and murine myocilin. Northern analysis of rat myocilin mRNA revealed substantial expression in the thyroid gland, as well as in the retina and muscle. No transcripts were detected in other endocrine glands, including the adrenal gland, pituitary, and testis. Myocilin may play an important role in thyroid function. Further study of the expression and role of myocilin in the thyroid is required. PMID- 10833335 TI - Is the Duchenne muscular dystrophy gene also an X-linked retinitis pigmentosa locus? AB - Deletion mutations and linkage mapping have localized an X-linked retinitis pigmentosa locus to Xp21, and a disease gene (RPGR) has been characterized. However, mutations have not been identified in most families expected to segregate the disease at this locus. Here, a retina-specific mRNA transcript from the Duchenne muscular dystrophy gene is identified. Based on these data, it is hypothesized that the Duchenne muscular dystrophy gene may represent a second Xp21 site at which retinitis pigmentosa mutations occur. PMID- 10833336 TI - Management of hospital outbreaks of gastro-enteritis due to small roundstructured viruses. AB - Small round structured viruses (SRSVs, Norwalk-like viruses, NLVs) are the most common cause of outbreaks of gastro-enteritis in hospitals and also cause outbreaks in other settings such as schools, hotels, nursing homes and cruise ships. Hospital outbreaks often lead to ward closure and major disruption in hospital activity. Outbreaks usually affect both patients and staff, sometimes with attack rates in excess of 50%. For this reason, staff shortages can be severe, particularly if several wards are involved at the same time. SRSVs may be spread by several routes: faecal-oral; vomiting/aerosols; food and water. Viruses may be introduced into the ward environment by any of these routes and then propagated by person-to-person spread. In an outbreak setting, the diagnosis can usually be made rapidly and confidently on clinical and epidemiological grounds, particularly if vomiting is a prominent symptom. By the time an SRSV outbreak has been recognized at ward level, most susceptible individuals will have been exposed to the virus and infection control efforts must prioritize the prevention of spread of infection to other clinical areas bycontainment of infected/exposed individuals (especially the prevention of patient and staff movements to other areas), hand-hygiene and effective environmental decontamination. This report of the Public Health Laboratory Service Viral Gastro-enteritis Working Group reviews the epidemiology of outbreaks of infection due to SRSVs and makes recommendations for their management in the hospital setting. The basic principles which underpin these recommendations will also be applicable to the management of some community based institutional outbreaks. PMID- 10833337 TI - Letters to the editor AB - The placement of MRSA carriers in private nursing homes: does information influence acceptance? Copyright 2000 The Hospital Infection Society. PMID- 10833338 TI - Letters to the editor AB - Workload may be related to the spread of MRSA and other infections Copyright 2000 The Hospital Infection Society. PMID- 10833339 TI - Experience of vancomycin-resistant enterococci in a children's hospital. AB - Vancomycin resistant enterococci (VRE) are increasingly important nosocomial pathogens. This paper describes our experience of the epidemiology and clinical impact of VRE in the two years since the occurrence of our first case of VRE infection. Following introduction of surveillance, gastrointestinal colonization with VRE was detected in 38.3% of Haematology/Oncology and 11.1% of Hepatology/Gastroenterology patients, but in only 2.3% of children in the Paediatric Intensive Care and 1.5% of children in the Renal Unit. Only five patients with gastrointestinal colonization subsequently developed clinical infection with VRE, giving an annual incidence of 7.5%. A further six children were colonized at extra-intestinal sites. Twelve children had clinical infections with VRE, of whom three (25%) died. Contamination of bedspaces was found in association with 2/3 (66.7%) children with extraintestinal colonization and 5/7 (71.4%) children with clinical infections, compared with 6/28 (21.4%) cases of gastrointestinal colonization. In the latter group, bedspace contamination was usually associated with widespread contamination of the ward with VRE and may have been the cause rather than the result of patients acquiring VRE. Originally we employed control measures based closely on the North American HICPAC guidelines, but our control strategy has since evolved in response to epidemiological and clinical observations. PMID- 10833340 TI - An evaluation of hospital cleaning regimes and standards. AB - A four-part study assessing cleanliness in up to 113 environmental surfaces in an operating theatre and a hospital ward is reported. Surfaces were assessed visually, using microbiological methods and ATP bioluminescence. Results from a preliminary random survey indicated variability in cleanliness. These results were then used to select sites for monitoring before and after routine cleaning, over a 14-day period. Using published microbiological and ATP specifications 70 and 76% of these sites were unacceptable after cleaning. Visual assessment was a poor indicator of cleaning efficacy with only 18% considered unacceptable. Sites most likely to fail in the ward were in the toilet and kitchen, areas which are frequently implicated in the spread of infectious intestinal disease. Operating theatre sites had lower ATP results but 61% of sites would be considered unacceptable. There was no significant difference in general microbiological or ATP results overall before and after routine cleaning. Although some important hand contact sites showed no significant difference, overall there was a significant decrease in staphylococcal and enterobacteria counts in the ward but not in the operating theatre after cleaning. The routine cleaning programmes used did not include a biocide and cleaning using a hypochlorite based sanitizer gave much lower values. The results are discussed in relation to infection control, cleaning audits and cleaning schedules: an integrated cleaning monitoring programme using ATP bioluminescence in conjunction with visual and microbiological assessments is recommended. PMID- 10833341 TI - Molecular investigation of Stenotrophomonas maltophilia isolates exhibiting rapid emergence of ticarcillin-clavulanate resistance. AB - The prevalence of Stenotrophomonas maltophilia resistance to ticarcillin clavulanate has dramatically increased in our hospital over the past few years. Sixty-six clinical isolates, collected from 61 patients, were tested for antibiotic susceptibility and genotyped by two different enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR) systems. Although two small outbreaks were identified in the medical intensive care unit, the remaining 58 strains generated 48 different PCR patterns. The rapid emergence of resistance in S. maltophilia occured in absence of an epidemic and was probably influenced by increasing-b-lactam use in our hospital. PMID- 10833342 TI - Epidemiology and susceptibility of serratia marcescens in a large general hospital over an 8-year period. AB - Over the 8 year period 1988-1995, 1367 isolates of Serratia marcescens were isolated from 582 patients on 12 different wards of a large Dublin hospital and were particularly associated with the surgical intensive care unit. The annual incidence was over 200 isolates from 1990 to 1992 but fell to below 100 following the opening in April 1992 of a replacement surgical hospital incorporating a new intensive care unit on the same site. The most common source of S. marcescens was sputum from patients. Strain identities were determined by serotyping and phage typing at least one isolate from each of 311 of the 582 patients. The results showed that a single epidemic strain of serotype O14:K14 was present in 69% of these patients, and persisted throughout the hospital for the whole of the eight year period. This strain was recovered from a variety of clinical specimens, including blood cultures. A minor outbreak involving a serotype O16:K28 strain also occurred and this strain also persisted from at least 1989 to 1994. Extensive surveillance failed to reveal an environmental source or faecal carriage. The likely mode of transmission appears to have been via staff hands from both symptomatic and asymptomatic patients acting as reservoirs of the organism, as has commonly been reported for this species. PMID- 10833343 TI - Repeated prevalence investigations on nosocomial infections for continuous surveillance. AB - In order to obtain an overview for the planning of further infection control activities, nine repeated prevalence studies were performed at monthly intervals. These occurred in the surgical units of eight medium-sized German hospitals. A total of 4984 surgical patients were investigated, the number of patients observed in each hospital varied from 365 to 913 patients, an average of 69.2 patients per prevalence study per hospital. A total of 212 nosocomial infections were found, the majority being surgical site (43.9%) and urinary tract infection (33.0%). The overall prevalence rate was 4.0%. More than four repeated investigations had only a minor influence on the 95% confidence intervals, and a follow-up of late microbiological reports increased the prevalence rate by only 7.5%. However, it was very useful to record the presence of urinary catheters on the prevalence day and also the preceding days; for instance, a device-associated prevalence of 7.8 urinary tract infections per 100 patients with urinary catheters was found on the day of investigation. In order to evaluate the situation in one's own surgical department by prevalence studies and for reasons of cost-effectiveness, the workload can be limited to four repeated studies in most hospitals. A further follow-up of later microbiological reports is not recommended, and it seems useful to concentrate on patients with indwelling devices. PMID- 10833344 TI - Occasional infection of hepatitis C virus occurring in haemodialysis units identified by serial monitoring of the virus infection. AB - To examine whether hepatitis C virus (HCV) infection still occurs in haemodialysis units even after a decrease in the number of blood transfusions and in those contaminated with HCV, we tested anti-HCV antibodies and HCV RNA in 142 patients from Japanese haemodialysis units, and examined the serial prevalence of anti-HCV antibodies in 86 patients from 1992 to 1997. A high prevalence of HCV infection was observed: 34 (23.9%) and 38 (26.8%) of the 142 patients were positive for serum anti-HCV antibodies and HCV RNA, respectively. These positive rates were related to the duration of haemodialysis. We found that five patients treated in the same unit seroconverted from 1993 to 1995. Four of the five patients had been treated at the same shift and were affected at the same time. Phylogenetic analysis of the hypervariable region 1 (HVR1) sequence of HCV from serum of these patients showed that three of the four patients' sequences were phylogenetically clustered and that two of the three were closely related. Thus, an occasional transmission of HCV occurred in the haemodialysis unit. The universal precautions by staff such as carefully changing gloves may be important in inhibiting spread of HCV because no instances of infection have been seen since the instigation of educational measures for unit staff. PMID- 10833345 TI - 'Now please wash your hands': the handwashing behaviour of final MBBS candidates. AB - The Handwashing Liaison Group has pointed out that "The failure of healthcare workers to decontaminate their hands reflects fundamentals of attitudes, beliefs and behaviours". Doctors are known to be poor at handwashing. This poor compliance may have its roots in a failure to learn this behaviour at medical college, where the influence of consultants and other role models may be critical. The handwashing behaviour of modern day medical students has not been previously studied. The Final MBBS Objective Structured Clinical Examination (OSCE) reflects learnt behaviours and attitudes of final year medical students 'absorbed' from role models within their training. We observed the handwashing behaviour of 187 candidates during the 1998 Final MBBS OSCE, at one clinical station, neurological examination of the lower limbs. Only 8.5% of candidates washed their hands after patient contact, although this figure rose to 18.3% with the aid of handwashing signs. These findings suggest that handwashing should become an educational priority. As student learning is highly focused by assessment (in-course or examination), we sug-gest that compliance with handwashing be built into undergraduate and Teaching Quality assessments with, for example, 'Hygiene marks' incorporated into OSCE or observed long case checklists. This study re-emphasizes the need for good clinical practice whenever teaching medical students. PMID- 10833346 TI - Bacterial flora on the white coats of medical students. AB - This study has demonstrated that the white coats of medical students are more likely to be bacteriologically contaminated at points of frequent contact, such as the sleeve and pocket. The organisms identified were principally skin commensals including Staphylococcus aureus. The cleanliness of the coat as perceived by the student was correlated with bacteriological contamination, yet despite this, a significant proportion of students only laundered their coats occassionally. This study supports the view that the students' white coat is a potential source of cross infection on the ward and its design should be modified in order to facilitate hand washing. Hospitals training medical students should consider taking on the burden of providing freshly laundered white coats for the students. PMID- 10833347 TI - Quantification of facial contamination with blood during orthopaedic procedures. AB - Operative surgery exposes the surgeon to possible blood-borne infections. Risks include pen-etrating injuries and conjunctival contact with infected blood. Visor masks worn during orthopaedic trauma procedures were assessed for blood contamination using computer analysis. This was found to be present on 86% of masks, of which only 15% was recognized by the surgeon intraoperatively. Of the blood splashes 80% were less than 0.6mm in diameter. We conclude that power instrumentation produces a blood particulate mist causing considerable microscopic, facial contamination which is a significant risk to the surgeon. PMID- 10833348 TI - Thermometers as a vehicle for transmission of extended-spectrum-beta- lactamase producing Klebisiella pneumoniae. PMID- 10833349 TI - Editorial policy PMID- 10833350 TI - Population viscosity and the evolution of altruism. AB - The term population viscosity refers to limited dispersal, which increases the genetic relatedness of neighbors. This effect both supports the evolution of altruism by focusing the altruists' gifts on relatives of the altruist, and also limits the extent to which altruism may emerge by exposing clusters of altruists to stiffer local competition. Previous analyses have emphasized the way in which these two effects can cancel, limiting the viability of altruism. These papers were based on models in which total population density was held fixed. We present here a class of models in which population density is permitted to fluctuate, so that patches of altruists are supported at a higher density than patches of non altruists. Under these conditions, population viscosity can support the selection of both weak and strong altruism. PMID- 10833351 TI - Bridging the gap: human diploid cell strains and the origin of AIDS. AB - Recent descriptions of the first human and chimpanzee cases of human immunodeficiency virus type 1 (HIV-1)-related retroviral infections dating from 1959 have stirred interest in the origin of AIDS. Although the theory of a chimpanzee origin of HIV-1 with cross-species transfer to man has now gained popularity, a more likely scenario is that chimps and humans were infected by an HIV-1 precursor virus derived from a contaminated poliovaccine. The reason for the rapidity and ease of cross-species transfer of this precursor virus has not been elucidated. We hypothesize that the poliovaccine was passaged in a human diploid cell strain. This simple manipulation allowed the retrovirus to adapt to human tissues and may have spawned the AIDS pandemic. PMID- 10833352 TI - Modeling complex neutrophil dynamics in the grey collie. AB - We have developed a mathematical model for the peripheral regulation of neutrophil production mediated by granulocyte colony-stimulating factor. We have used that model to show that the pattern of neutrophil oscillations in nine grey collies is consistent with the hypothesis that cyclical neutropenia is due to an oscillatory stem cell input to the neutrophil regulatory system, and not due to autonomous oscillations in the peripheral neutrophil regulatory system. In the process of interfacing our model with the laboratory data, we have estimated parameters for the peripheral neutrophil control system consistent with higher than normal apoptotic cell loss within the recognizable neutrophil precursors. This is in agreement with other experimental data. Our estimated model parameters also predict that the peripheral neutrophil production system is globally stable in the grey collies we studied. This further supports our hypothesis that the origin of the oscillatory behavior in cyclical neutropenia is in the stem cell population, consistent with other clinical and experimental evidence. PMID- 10833353 TI - A substrate switch: a new mode of regulation in the methionine metabolic pathway. AB - We propose a simple mathematical model of liver S -adenosylmethionine (AdoMet) metabolism. Analysis of the model has shown that AdoMet metabolism can operate under two different modes. The first, with low metabolic rate and low AdoMet concentration, serves predominantly to supply the cell with AdoMet, the substrate for various cellular methylation reactions. The second, with high metabolic rate and high AdoMet concentration, provides an avenue for cleavage of excess methionine and can serve as a source of cysteine when its increased synthesis is necessary. The switch that triggers interconversion between the "low" and "high" modes is methionine concentration. Under a certain set of parameters both modes may coexist. This behavior results from the kinetic properties of (i) the two isoenzymes of AdoMet synthetase, MATI and MATIII, that catalyse AdoMet production; one is inhibited by AdoMet, whereas the other is activated by it, and (ii) glycine- N -methyltransferase that displays highly cooperative kinetics that is different from that of other AdoMet-dependent methyltransferases. Thus, the model provides an explanation for how different cellular needs are met by regulation of this pathway. The model also correctly identifies a critical role for glycine N -methyltransferase in depleting excess methionine in the high mode, thus avoiding the toxicity associated with elevated levels of this essential amino acid. PMID- 10833354 TI - Length distribution of the peptidoglycan chains in the sacculus of Escherichia coli. AB - The stress-bearing fabric of bacteria is made of peptidoglycan. This crosslinked fabric is formed from disaccharide pentapeptide units that are transported through the cytoplasmic membrane and then polymerized in two directions: (i) to form oligoglycan chains; and (ii) to cross link these chains by tail-to-tail bonds from the muropeptides to the protruding peptides of other chains. The distribution of the glycan chain lengths is reminiscent of the "most probable distribution of polymer chemistry. Of course, the process is more complex than solely the random addition of units to growing chains. The complexity precludes mathematical analysis, but computer modeling of the Monte Carlo type is capable of including a range of possibilities. At each time point a specified number of disaccharides are singly added to the muramic acid residue ends of existing chains chosen at random. The transfer is in exchange for the cleavage of pyrophosphate bactoprenol that transported the disaccharide pentapeptide through the membrane. The progam then selects, again at random, which chain to cleave and between which two disaccharides of the chain the cleavage event is to occur. The cleavage generates an N -acetyl 1,6 anhydro-muramic acid end and a non-reducing N -acetyl glucosamine end. The simulation can be modified so that the program does not cleave off a disaccharide next to either end of the chain. Comparisons are shown with the experimental results of Obermann & H]oltje (1994. Microbiology140, 79-87.) They obtained their data by taking the results with normal growing cells and subtracting the similar data from minicells to estimate the chain length distribution in the cylinder part of the cell. In its most basic form the computer simulation has only one fitted parameter, K, which is the number of disaccharides added to the murein for every internal cleavage event. In this form the fitting to the experimental results is poor. One possible reason for this is that the tension on the chains, and therefore the probability of being cleaved by autolysins varies with orientation of the chain on the cylinder surface. It is well known that the tension in the cylindrical wall is twice as large in the circumferential direction as in the axial one, so one class would consist of those chains aligned longitudinally, subject to lower stress, and would have a higher energy of activation for autolysis than chains aligned circumferentially. A good fit is obtained on the assumption that there are only two classes of chains; one more likely to be cleaved than the other. The key point is that only two processes: adding of disaccharide pentapeptides at random to glycan chains and cleavage between the disaccharides at random, together with the assumption that the wall is less easily hydrolysed in the axial direction is sufficient to account for the experimental distribution. PMID- 10833355 TI - Hawks, doves, and mixed-symmetry games. AB - The hawk-dove game has proved to be an important tool for understanding the role of aggression in social interactions. Here, the game is presented in a more general form (GHD) to facilitate analyses of interactions between individuals that may differ in "size", where size is interpreted as a surrogate for resource holding power. Three different situations are considered, based on the availability and use of information that interacting individuals have about their sizes: the classical symmetric case, in which no information about sizes is used, the asymmetric case, in which the individuals know their relative sizes and thus their chances of prevailing in combat, and a mixed-symmetry case, in which each individual only knows its own size (or only knows its opponent's size). I describe and use some recently developed methods for multitype games-evolutionary games involving two or more categories of players. With these methods and others, the evolutionarily stable strategies (ESSs) that emerge for the three different cases are identified and compared. A proof of the form and uniqueness of the ESS for the mixed-symmetry case is presented. In this situation, one size category at most can play a mixed strategy; larger individuals are aggressive and smaller individuals are not. As the number of size categories approaches infinity and the size distribution becomes continuous, there is a threshold size, above which all individuals are aggressive, and below which they are not. PMID- 10833356 TI - Delay model of the circadian pacemaker. AB - We present a simple and realistic model of the circadian pacemaker that can be interpreted in molecular terms. The model, which consists of a single time-delay differential equation, simulates the expression of a generic clock protein that inhibits its own expression through a feedback mechanism. Despite its simplicity, this model fulfils most of the necessary characteristics of a realistic representation of natural circadian clocks: robust and stable oscillations with circadian free-running periods, typical phase response curves and entrainment to environmental zeitgebers. The present model reduces the molecular mechanism necessary to sustain stable oscillations to its bare bones, suggesting that the essential factor is the time-delayed negative feedback of the oscillating protein on its own expression. PMID- 10833357 TI - The effect of inspired oxygen concentration on the ventilation-perfusion distribution in inhomogeneous lungs. AB - The coupled conservation of mass equations for oxygen, carbon dioxide and nitrogen are written down for a lung model consisting of two homogeneous alveolar compartments (with different ventilation-perfusion ratios) and a shunt compartment. As inspired oxygen concentration and oxygen consumption are varied, the flux of oxygen, carbon dioxide and nitrogen across the alveolar membrane in each compartment varies. The result of this is that the expired ventilation perfusion ratio for each compartment becomes a function of inspired oxygen concentration and oxygen consumption as well as parameters such as inspired ventilation and alveolar perfusion. Another result is that the "inspired ventilation-perfusion ratio and the "expired ventilation-perfusion ratio differ significantly, under some conditions, for poorly ventilated lung compartments. As a consequence, we need to distinguish between the "inspired ventilation-perfusion distribution, which is independent of inspired oxygen concentration and oxygen consumption, and the "expired ventilation-perfusion distribution, which we now show to be strongly dependent on inspired oxygen concentration and less dependent oxygen consumption. Since the multiple inert gas elimination technique (MIGET) estimates the "expired ventilation-perfusion distribution, it follows that the distribution recovered by MIGET may be strongly dependent on inspired oxygen concentration. PMID- 10833358 TI - Cell kinetics in a tumour cord. AB - In some tumours, the viable cells grow around blood vessels forming cylindrical structures called tumour cords, which are surrounded by regions of necrosis. In the present paper, we propose a mathematical model for the cell kinetics in a tumour cord at the stationary state. Both proliferating cells and quiescent cells are considered, and the proliferating cell population is structured by age. Cell migration towards cord periphery is accounted for from a continuum viewpoint. The age distribution of proliferating cells, the fraction of cells in S phase, the growth fraction and the velocity along the cord radius are computed. The predictions of the model are compared with literature data obtained from two experimental rat hepatomas. The model was used to compute the profile of the oxygen tension within the cord. Possible modifications and extensions are also presented. PMID- 10833359 TI - Mathematical elements of attack risk analysis for mountain pine beetles. AB - Three different mathematical approaches are combined to develop a spatial framework in which risk of mountain pine beetle (MPB) attack on individual hosts may be assessed. A density-based partial differential equation model describes the dispersal and focusing behavior of MPB. A local projection onto a system of ordinary differential equations predicts the consequences of the density equations at individual hosts. The bifurcation diagram of these equations provides a natural division into categories of risk for each host. A stem competition model links host vigor to stand age and demographics. Coupled together, these models illuminate spatial risk structures which may also shed light on the role of climatic variables in population outbreaks. Preliminary results suggest that stand microclimate has much greater influence on risk of attack than host vigor and stand age. PMID- 10833360 TI - Is the United States country zero for the First-World AIDS epidemic? AB - The incidence for AIDS per hundred thousand is several times higher in the United States than in the rest of the First World. Earlier work by Thompson (1984, 1989a, b, 1990, 1998) indicated that a relatively small proportion of gay males frequenting the bathhouses in the United States, drove AIDS over the epidemiological threshold in the U.S. It is shown that the rate of growth of AIDS is essentially the same for the United States and other First-World countries. An argument is advanced, based on WHO AIDS data, to the effect that it is contact with the pool of infectives in the United States that drives the epidemic in other First-World Countries. PMID- 10833361 TI - Ancestral processes in population genetics-the coalescent. AB - A special stochastic process, called the coalescent, is of fundamental interest in population genetics. For a large class of population models this process is the appropriate tool to analyse the ancestral structure of a sample of n individuals or genes, if the total number of individuals in the population is sufficiently large. A corresponding convergence theorem was first proved by Kingman in 1982 for the Wright-Fisher model and the Moran model. Generalizations to a large class of exchangeable population models and to models with overlying mutation processes followed shortly later. One speaks of the "robustness of the coalescent, as this process appears in many models as the total population size tends to infinity. This publication can be considered as an introduction to the theory of the coalescent as well as a review of the most important "convergence to-the-coalescent-theorems. Convergence theorems are not only presented for the classical exchangeable haploid case but also for larger classes of population models, for example for diploid, two-sex or non-exchangeable models. A review like summary of further examples and applications of convergence to the coalescent is given including the most important biological forces like mutation, recombination and selection. The general coalescent process allows for simultaneous multiple mergers of ancestral lines. PMID- 10833362 TI - Motional instabilities in prey-predator systems. AB - Differential fluxes can destabilize the locally stable stationary density distributions in interaction systems with diffusion, advection, and/or locomotion of chemical or biological species. By this method they can cause the formation of stationary or travelling spatial structures. Different scenarios of this general mechanism of spatio-temporal pattern formation in reaction-diffusion-advection systems will be demonstrated, using a simple two-species predator-prey system as an example model. PMID- 10833363 TI - Spatial and temporal patterns of expression of messenger RNA for insulin-like growth factors and their binding proteins in the placenta of man and laboratory animals. AB - To better understand the role of the insulin-like growth factors (IGF-I and -II) and their binding proteins (IGFBPs 1-6) in placental development and function, it is important to review similarities and differences between species in expression of the respective mRNAs. In human placenta, IGF-II mRNA is expressed in chorionic mesoderm and first trimester villous cytotrophoblast, but not in syncytiotrophoblast. In contrast, in rhesus monkey placenta, IGF-II mRNA is expressed in syncytiotrophoblast but not in chorionic mesoderm. IGFBP-3 mRNA is present in the chorionic mesoderm of placental villi from both these species and may modulate IGF-II action through a paracrine mechanism. In rodent placentae, IGF-II mRNA is expressed both in fetal mesoderm and in the trophoblast of the placental labyrinth. In guinea pig, where IGFBP-5 mRNA is expressed in the marginal and interlobular syncytium and IGF-II mRNA in the labyrinth, interaction between IGF-II and IGFBP-5 mRNA may be involved in vascularization of the placenta by fetal vessels. In sheep placenta, IGF-II mRNA is expressed, not in the trophoblast layer, but in the fetal mesoderm immediately adjacent to it. In the basal plate of human, rhesus monkey and baboon placentae, extravillous trophoblasts express IGF-II mRNA and uterine decidual cells IGFBP 1-6 mRNAs. The inference is that there is interaction between IGF-II and IGFBPs at the maternal fetal interface of the primate placenta during trophoblast invasion and decidualization. IGFBP-1 expressed by the decidua may also interact with alpha(5)beta(1)integrin expressed by the extravillous trophoblast. The placentae of rodents are also of the invasive type. Glycogen cells of the mouse placenta are analogous with human extravillous trophoblast and express IGF-II mRNA. However, expression of IGFBP mRNAs in the mouse, as in the guinea pig, is confined to non-decidualized endometrium and myometrium. IGF-II mRNA is strongly expressed by trophoblasts invading uterine vessels in human and guinea pig placentae. Interactions probably occur between IGF-II expressed by these trophoblasts and IGFBPs expressed in the vessel walls. However, it is possible that IGFBPs expressed by maternal vessels are associated with processes that are independent of trophoblast invasion. Thus, IGFBP-3 mRNA is highly expressed in the maternal blood vessels of the non-deciduate sheep placenta. Findings to date highlight the diversity in the expression of the IGF system among placentae of man and different laboratory animals, and even between closely related species. Comparative studies will continue to be required to understand the functional role of IGFs and IGFBPs in each species. PMID- 10833364 TI - Pregnancy-induced de-differentiation of media smooth muscle cells in uteroplacental arteries of the guinea pig is reversible after delivery. AB - The majority media of smooth muscle cells of uteroplacental arteries of the guinea pig is not destroyed during trophoblast invasion. Rather, most of these cells de-differentiate during pregnancy-induced arterial dilatation, forming a population of mesenchyme-like myoblasts ready to reconstitute the media after birth. We have studied the re-differentiation of these cells after delivery by means of transmission electron microscopy and immunohistochemistry using antibodies against a panel of cytoskeletal proteins. The data reveal that post partum re-differentiation of the media myoblasts starts immediately after birth where some of the invasive trophoblast cells are still present. The process of re differentiation is completed at day 8 after parturition. Post partum re differentiation can be subdivided into two steps: until day 5 after parturition, the central parts of the media are reconstituted out of the reservoir of vimentin positive myoblasts by stepwise acquisition of desmin, alpha-smooth muscle actin, gamma-smooth muscle actin and smooth muscle myosin. Only thereafter the same re differentiation takes place in the peripheral parts of the media. On the ultrastructural level immunohistochemical re-expression of cytoskeletal proteins is accompanied by reconstitution of the intracellular contractile apparatus. The data support our earlier notion that the majority of media smooth muscle cells in the guinea pig uterus does not degenerate during trophoblast-invasion but rather de-differentiate temporarily. PMID- 10833365 TI - Chronic hypoxia diminishes pregnancy-associated DNA synthesis in guinea pig uteroplacental arteries. AB - Enlargement of the uterine artery (UA) during pregnancy is diminished in women residing at a high altitude. We asked whether chronic hypoxia alters the rise in DNA synthesis in uteroplacental vessels and, if so, whether the reduction is related to the intrauterine growth retardation (IUGR) observed under conditions of chronic hypoxia. We used bromodeoxyuridine (BrdU) labelling to measure DNA synthesis in all vascular layers of the UA, mesometrial arteries (MA), thoracic aorta and mesenteric artery of guinea pigs, residing throughout pregnancy at a low (1600 m) or high (3962 m) altitude. Pregnancy increased DNA synthesis throughout the UA at both altitudes, yet the maximal value was less at high than low altitude (P<0.05). Likewise, pregnancy increased DNA synthesis throughout the MA, yet at high altitude pregnancy elevated levels returned to non-pregnant values after 42 days of gestation, whereas at low altitude DNA synthesis continued to be elevated until near term. Fetal weights were lower (P=0.01) and placental/fetal weight ratios tended to be greater in high than low altitude, near term pups (P = 0.09). We conclude that a diminished growt response by the uteroplacental vasculature to pregnancy may contribute to the previously reported reduced uterine artery blood flow and resulting IUGR at high altitude. PMID- 10833366 TI - Expression and regulation of vascular endothelial growth factor in a first trimester trophoblast cell line. AB - Embryo implantation and development are critically dependent upon the spatial and temporal regulation of angiogenesis and localized vascular permeability. A key mediator of these effects is the endothelial cell mitogen vascular endothelial growth factor (VEGF). VEGF has been shown to promote endometrial vascular permeability, fetal vasculogenesis and placental, fetal and maternal angiogenesis. However, the mechanism through which this regulation occurs in the placenta is poorly understood. This study was conducted to determine if the pro angiogenic cytokines, TNF-alpha and TGF-beta1, affect VEGF expression in human first trimester trophoblasts. Culture of a first trimester trophoblast cell line (HTR-8/SVneo), in the presence of either TNF-alpha or TGF-beta1, resulted in the expression of significant levels of VEGF in culture. The trophoblast cell line also showed a time-dependent and a dose-dependent increase in VEGF mRNA levels when cultured in the presence of either TNF-alpha or TGF-beta1. These results suggest that both TNF-alpha and TGF-beta1 may regulate the production of VEGF in early gestational trophoblasts and may therefore serve to modulate placental vascular permeability and angiogenesis that are necessary for embryo implantation and placentation. PMID- 10833367 TI - Establishment of a pure vascular endothelial cell line from human placenta. AB - Endothelial cells (EC) from various sectors of the circulatory system have distinct characteristics, some of which have only been identified in cultures upon their isolation from specific organs or tissues. Cultured vascular EC, derived from the human placenta (HPEC), may be helpful for studying their specific function in the fetoplacental unit, such as in the control of maternofetal traffic. In this paper we report an improved method for isolation, purification and culture of HPEC, that implies an enzymatic perfusion of the term placenta, followed by separation of resulting cells on a Percoll density gradient. The inoculated starting suspension was purified by a two-step selection procedure, based on differential trypsinization, leading to a pure population of about 8x10(7)cells/placenta, with 2.7-3.4 population doublings. The average population doubling time during eight passages was 60-65 h and the life span of HPEC was approximately 45-50 population doublings. The cell morphology at optical and electron microscopical level revealed a good differentiation of HPEC, which were endowed with numerous plasmalemmal vesicles (caveolae) and Weibel-Palade bodies. The transendothelial electrical resistance of the HPEC monolayer varied between 22 and 52 Ohm/cm(2). The cultures were mycoplasma free, as revealed by fluorescence microscopy using DNA dyes and the polymerase chain reaction (PCR). The negative immunofluorescent reaction for keratin confirmed that the HPEC were not contaminated with either type of placenta cells, as syncytiotrophoblast. Cultured HPEC demonstrated a strong reaction for von Willebrand factor antigen (by fluorescence microscopy), took up AcLDL-DiI and expressed active angiotensin converting enzyme. These characteristics substantiate the endothelial nature of cultured cells. The interactions with different lectins (BS-I, SBA, RCA, UEA and WGA) assessed by fluorescence microscopy and blotting reveal a strong reaction of HPEC with UEA and a negligible reaction with BS-I lectin. WGA lectin displayed a marked fluorescence staining in subconfluent HPEC, and at the level of intracellular clefts in post-confluent cultures. IN CONCLUSION: (i) we have obtained a pure line of cultured EC originating from the human placental venous side of the circulatory tree; (ii) the cells have the general characteristics and markers ascribed to EC; (iii) as opposed to large human placental vessels, HPEC do not react to BS-I lectin and, unlike human umbilical vein EC, have a much higher proliferation rate and a long lifespan; (iv) HPEC expressed a characteristic glycosylated coat particularly rich in alpha- L -fucose and beta GlcNAc containing glycocompounds. PMID- 10833368 TI - Haem oxygenase activity in human umbilical cord and rat vascular tissues. AB - Carbon monoxide (CO) has been shown to affect vascular tone in smooth muscle cells and thus, may regulate regional or systemic blood pressure as well as fetoplacental vascular tone and fetal blood delivery. To assess the potential of vascular tissue to produce CO, we determined haem oxygenase (HO) activity through in vitro quantitation of CO production with gas chromatography and its inhibition by 33-66 microm of chromium mesoporphyrin (CrMP) in homogenate preparations of rat aorta and vena cava and human umbilical cord tissues. We compared these results to HO activity in rat heart and liver. We also discuss normalization of HO activity on a per mg protein as well as per g fresh weight (FW) tissue basis. We found that both rat vascular tissue HO activities (per g FW) were equal, but greater than that of heart (x3) and less than that of liver (x0.2). For human cord tissues, HO activities of artery and vein were equal, but greater than that of Wharton's jelly. Also, HO activity in rat vascular tissues was 3x greater than that of the human cord tissues. HO activity was completely inhibited by CrMP in rat heart (90 per cent) and liver (96 per cent), but incompletely (50-66 per cent) in both rat and human vascular tissues. We established that it is unlikely that other non-haem CO-generating processes account for this unique insensitivity of HO to CrMP inhibition. In fact, high concentrations of other potent metalloporphyrin inhibitors affected vascular tissue HO even less. We found that the degree of in vitro HO inhibition appeared to be related to the concentration of haem in the reaction medium. We conclude that the presence of HO activity in cord tissues supports the possibility that CO plays a role in fetoplacental blood flow regulation. PMID- 10833369 TI - Pregnancy-dependent expression of leukaemia inhibitory factor (LIF), LIF receptor beta and interleukin-6 (IL-6) messenger ribonucleic acids in the porcine female reproductive tract. AB - Leukaemia inhibitory factor (LIF) and interleukin-6 (IL-6) are candidate embryo maternal signalling molecules which are present within the uterine luminal micro environment. We examined the relative expression of the mRNAs encoding LIF and IL 6, as well as the LIF-binding subunit (LIFR-beta) of the LIF receptor and, as a potential downstream cytokine-responsive gene, beta(2)-microglobulin (beta(2)m), in porcine peri-implantation conceptuses, and in placenta and endometrium during early and mid-pregnancy. Peri-implantation spherical and filamentous conceptuses expressed LIFR-beta and beta(2)m mRNAs with no LIF mRNA present. Rapid development in days 11/12 spherical conceptuses to the filamentous stage was accompanied by transiently increased IL-6 gene expression. The corresponding endometrium, in contrast, expressed LIF in addition to these other mRNAs. LIFR beta, IL-6 and beta(2)m, but not LIF mRNAs, were expressed in the Jag-1 cell line, an in vitro model for porcine day 14 trophoblast. The greatest steady-state amounts of LIF, LIFR-beta and IL-6 mRNAs in both the endometrium and placenta were evident at the post-implantation stages (days 30 and 60>day 18 of pregnancy). Treatment of porcine endometrial explants with human recombinant (hr)LIF or hrIL-6 resulted in no change in, or diminished, the presence of endometrial beta(2)m mRNA, respectively. Addition of LIF to peri-implantation conceptus explant cultures, in contrast, induced beta(2)m mRNA synthesis. These results highlight the potential importance of both the endometrium and placenta as sources, as well as targets, of these cytokines throughout pregnancy. Cytokine modulation of beta(2)m, a known in vitro mitogen, may constitute one mechanism for local control of trophoblast and endometrial proliferation. PMID- 10833370 TI - Production of interleukin (IL)-6 and IL-8 by a choriocarcinoma cell line, BeWo. AB - To clarify the biological and pathological features of choriocarcinoma, we evaluated the in vitro production of cytokines by a choriocarcinoma cell line, BeWo. We measured the concentration of interleukin (IL)-6 and IL-8 in the culture media of BeWo cells after stimulation with various modulatory agents of cytokine expression by enzyme-linked immunosorbent assays. Northern blot analysis was used to examine the expression of IL-6 and IL-8 mRNA in these cells. A weak expression of IL-6 and IL-8 was detected in unstimulated BeWo cells by both methods. IL-6 transcription and secretion were dose-dependently enhanced by stimulation with IL 1beta, tumour necrosis factor (TNF)-alpha, transforming growth factor (TGF)-beta1 and 12-O-tetradecanoyl phorbol-13-acetate (TPA). Forskolin, lipopolysaccharide and interferon-gamma had no effect on these cytokines production. The TNF-alpha induced secretion of IL-6 was inhibited by dexamethasone. The TPA-induced production of IL-6 was inhibited by 1-(5-isoquinolinylsulfonyl)-2 methylpiperazine and sphyngosine, suggesting the involvement of a protein kinase C-dependent pathway. Levels of IL-8 mRNA and protein were also dose-dependently increased by stimulation with IL-1beta, TNF-alpha and TPA. In contrast to IL-6, the expression of IL-8 was not affected by TGF-beta1. It is suggested that, in addition to the production of steroidal and non-steroidal hormones, these cytokines may serve as part of a cytokine network that modulates the proliferation and angiogenesis of choriocarcinomas. PMID- 10833371 TI - Expression of BCL-2, BAX and BAK in the trophoblast layer of the term human placenta: a unique model of apoptosis within a syncytium. AB - The regulation of apoptosis in the syncytiotrophoblast is of particular interest because this is the only true syncytial epithelium in human cell biology. Nuclei characteristic of apoptotic cells have been localized to this syncytium especially in association with fibrin-containing fibrinoid deposits. The factors responsible for regulating cell death-like features in the trophoblast syncytium are unknown. We tested the hypothesis that fibrin was required for trophoblast apoptosis. TUNEL (terminal deoxynucleotidyltransferase-mediated dUTP end labelling) staining to detect DNA fragmentation typical of apoptosis was performed in term human placentae revealing labelled nuclei associated with fibrin-type fibrinoid, as well as labelled nuclei in discrete areas of syncytiotrophoblast without fibrin. We also hypothesized that members of the BCL 2 family of apoptosis-associated proteins contribute to the regulation of syncytiotrophoblast apoptosis. To identify members of this protein family that might regulate trophoblast apoptosis, we assessed expression of three important members of the bcl-2 gene family. We used immunohistochemistry with monoclonal antisera against human BCL-2 and polyclonal antisera against human BAX and BAK to study paraffin-embedded sections of human term placentae (n=5) from uncomplicated pregnancies. The anti-apoptotic BCL-2 protein was expressed throughout the syncytium of normal villi with much less staining in cytotrophoblast. Staining was also seen adjacent to fibrin deposits and in syncytium overlying fibrin deposits. Expression of the pro-apoptotic BAX protein was undetectable in the syncytiotrophoblast, was expressed in rare cytotrophoblast and was prominent in connective tissue and perivascular cells within the villous core. Localization of a second pro-apoptotic protein, BAK, revealed immunoreactivity in isolated areas of intact syncytium of normal villi. Additionally, fibrin deposits were associated with intense BAK staining in both syncytiotrophoblast and cytotrophoblast. From these data, we speculate that modulation of BAK expression is one factor regulating apoptosis in human trophoblast. PMID- 10833372 TI - Fatty acid transport regulatory proteins in the developing rat placenta and in trophoblast cell culture models. AB - The placenta forms a selective barrier that is able to transport nutrients that are of critical use to the fetus. Delivery of essential fatty acids to the fetus is dependent upon transplacental transport and provides the backbone for the biosynthesis of biological membranes, myelin and various signalling molecules. The primary objective of this research was to elucidate the expression patterns of genes that regulate fatty acid transport across the placenta. Several fatty acid transport regulatory genes have been identified in the rat including; cytoplasmic heart fatty acid binding protein (hFABP), plasma membrane fatty acid binding protein (FABPpm), fatty acid translocase (FAT) and fatty acid transport protein (FATP). In this study, we have elucidated temporal and spatial expression patterns for these genes in the rat placenta and in cell culture models of the rat placenta by Northern blot, RT-PCR, Western blot and/or by in situ hybridization analyses. Expression of hFABP was specific to the labyrinth zone, the main barrier and site of transplacental transport in the rat placenta. In addition, the levels of hFABP expression increased with gestational age, suggesting a growing requirement for fatty acid transport with advancing stages of pregnancy. FABPpm, FAT and FATP are expressed in both the junctional and labyrinth zones of the rat placenta. FAT was predominantly localized to the labyrinth zone by in situ hybridization analysis. The placental cell expression patterns of the genes involved in fatty acid transport were supported by our observations of HRP-1 (labyrinth zone) and Rcho-1 (junctional zone) trophoblast cell culture models. Given their cell surface location, we predict that FABPpm, FAT and FATP potentially participate in placental fatty acid uptake. The predominant expression of hFABP and FAT in the labyrinth zone of the chorioallantoic placenta implicates hFABP and FAT in the transplacental movement of fatty acids from maternal to fetal compartments. PMID- 10833373 TI - Surface expression of HLA-C antigen by human extravillous trophoblast. AB - In this paper definitive evidence that the classical class I product, HLA-C, is expressed on the surface of normal trophoblast cells is provided. HLA-C transcripts were sequenced from cDNA isolated from first trimester trophoblast cells obtained by flow cytometric sorting. Both paternal and maternal alleles were transcribed. HLA-C proteins were demonstrated by biochemical analysis and found on the cell surface in association with beta(2)-microglobulin. Upregulation of cell surface HLA-C but not HLA-G expression after interferon (IFN)-gamma treatment was demonstrated by flow cytometric analysis. Immunohistology has confirmed HLA-C is expressed by all extravillous subpopulations in vivo. The question of whether trophoblast HLA-C molecules interact with decidual NK cells expressing killer Ig-like receptors (KIR) has also been addressed. Our results demonstrate that extravillous trophoblast expresses at least two HLA class I molecules, HLA-G and HLA-C on the cell surface. PMID- 10833374 TI - Maternal blood coagulation factor XIII is associated with the development of cytotrophoblastic shell. AB - We analysed the early implantation tissues of normal women and of a patient with congenital factor XIII deficiency in order to study the role of maternal subunit A of factor XIII (XIIIA) in the development of extravillous cytotrophoblast. The patient had received adequate administration of factor XIIIA concentrate only up to 7 weeks of gestation (wG). Her pregnancy was maintained until the latter half of 8 wG, but was terminated by intrauterine fetal death at 9 wG. Immunohistochemical staining of cytokeratin, XIIIA and subunit S of factor XIII was performed in the early implantation tissues of normal women and of this patient. Numerous well-formed cytotrophoblastic shells and Nitabuch's layers were detected in implantation tissues at 7-8 wG in normal women, and XIIIA was present in the intercellular space in well-formed cytotrophoblastic shells, while the cytotrophoblastic shells and Nitabuch's layers in this patient's implantation tissue were poorly-formed. Furthermore, XIIIA was not detected around them. It is suggested that when the maternal plasma activity of factor XIII is low, the concentration of XIIIA at the placental bed is also low, leading to the insufficient formation of cytotrophoblastic shell and therefore an increased probability of miscarriage in patients with congenital factor XIII deficiency. PMID- 10833375 TI - The characterization of human amnion epithelial and mesenchymal cells: the cellular expression, activity and glucocorticoid regulation of prostaglandin output. AB - The amnion, a single layer of epithelial cells (EC) overlying layers of mesenchymal cells (MC) has been identified as a source of intrauterine prostaglandins (PG). The objectives of the present study were: (1) to establish a technique for the isolation and culture of pure amnion EC and MC preparations, (2) to characterize the cellular expression of PGHS-II and PGHS activity within these separated amnion cells and (3) to characterize the pattern of glucocorticoid stimulation of these separated amnion cells. Term gestation human amnion was collected after elective caesarean section or vaginal delivery. A trypsin digestion was used to isolate EC and a mechanical digestion and collagenase dispersion was used to isolate MC. Following 48 or 96 h in culture, cells were incubated for 24 h in the presence or absence of 1 microm arachidonic acid and treated with cortisol (F: 10-1000 nm) or 1 microm dexamethasone (DEX). Cell types were identified by immunohistochemistry (IHC). Immunoreactive PGHS-II (ir-PGHS-II) and glucocorticoid receptor (ir-GR) were localized by IHC. PGHS activity was measured as PGE(2)output determined by radioimmunoassay. Mean PGE(2)production by MC at 72 h was 22-fold greater (P<0.05) and at 120 h was 32 fold greater (P<0.03) than PGE(2)output by EC. Administration of arachidonic acid stimulated a 5.0-fold increase in PGE(2)output (P<0.0002) by EC after 72 h and a 3.6-fold increase (P<0.05) after 120 h but did not alter MC PGE(2)output. Despite exogenous substrate, EC PGE(2)output remained significantly less than PGE(2)output by MC. There was no difference in PG production by EC and MC with the onset of labour. Ir-GR expression was found in both EC and MC. F and/or DEX with and without arachidonic acid (AA) stimulated PGE(2)output by EC. Only DEX and not F increased PGE(2)output by MC. These data suggest that relatively pure EC and MC preparations can be established from amnion. PG output and its regulation appears to differ within these two amnion cell types, dependent upon (1) substrate availability and (2) the regulation of PGHS activity. PMID- 10833376 TI - Risk factors for neonatal intraventricular haemorrhage in spontaneous prematurity at 32 weeks gestation or less. AB - In this study we aimed to establish which clinical and histopathological factors are associated with early-onset neonatal intraventricular haemorrhage (IVH) in non-iatrogenic preterm delivery before 32 weeks of gestation. We retrospectively reviewed all singleton pregnancies delivered before 32 weeks of gestation after spontaneous onset of preterm labour or preterm membrane rupture during the period January 1993 to June 1997. Clinical and histopathological data in cases with IVH diagnosed at neonatal cranial ultrasound within 72 h of birth (n = 17) were compared with those of neonates not experiencing this complication (non-IVH) (n = 54). Histological lesions analysed were those of acute inflammation and those on a uteroplacental vascular basis. Statistical methods included the Wilcoxon rank sum test, Fisher's exact test, and logistic regression analysis. A P<0.05 was considered significant.IVH and non-IVH groups were not significantly different in birthweight, gestational age at delivery, cord pH at birth, rates of 5-min Apgar score below 7, caesarean delivery, diagnosis of clinical chorioamnionitis or antenatal administration of steroids. Respiratory distress syndrome was more frequently diagnosed in the IVH than non-IVH group (64 per cent versus 33 per cent, P=0.02). Placental acute inflammatory or uteroplacental vascular lesions were present in 100 per cent of IVH neonates versus 22 per cent of non-IVH cases (P<0.001). Logistic regression analysis demonstrated that only respiratory distress syndrome (P = 0.04) and histological evidence of acute placental inflammation (P = 0.02) were significantly and independently associated with IVH. Histopathological evidence of acute inflammatory placental lesions is the best predictor of occurrence of neonatal IVH. PMID- 10833377 TI - Diminished expression of placental isoferritin p43 component in first trimester abnormal pregnancies. AB - Human placental isoferritin (PLF) is a sub-type of human ferritin mainly composed of a 43 kD protein, which has an immunosuppressive activity and may be involved in the downregulation of the maternal immune system during pregnancy. The aim of this study was to evaluate the distribution of p43 in the placental tissue of abnormal first trimester pregnancies. Samples of villous and decidual tissues were collected between 7 and 12 weeks' gestation from 28 missed abortions and eight complete moles. Samples of placental tissue from 20 normal pregnancies of similar gestational age were used as controls. The localization of p43 was determined by immunohistochemical techniques using CM-H9 monoclonal antibody. Compared to controls, specific p43 immunoreactivity was low in the villous syncytiotrophoblast of missed abortions and absent from all villous cellular types in complete moles. These findings correlate well with the low level of maternal serum PLF found previously in early pregnancy failures and molar gestation. This suggests that PLF may be involved in the pathogenesis of early pregnancy disorders related to an abnormal placentation. PMID- 10833378 TI - Identification of Gaucher cells in the chorionic villi associated with recurrent hydrops fetalis. AB - Recurrent non-immune hydrops fetalis has rarely been reported. In order to detect the risk of recurrence in a subsequent pregnancy, one should carefully consider the possibility of an inborn error of metabolism. In such cases, placental examination may be useful in detecting such metabolic storage disorders in the fetus, which usually present as vacuolization of placental cells. We describe a rare case of recurrent hydrops that was detected by placental examination. Through light microscopy, electron microscope (EM) studies and beta glucocerebrosidase activity the disease was identified as Gaucher's disease. PMID- 10833379 TI - Placental haemozoin and malaria in pregnancy. AB - Malaria infections in pregnant women cause poor birth outcomes. Malaria pigment (haemozoin) accumulates in the placenta within macrophages and extracellularly, but its pathological significance is not understood. In order to study the potential role of haemozoin in malaria pathogenesis, we enrolled primigravid women at a Malawian government antenatal clinic and followed them through delivery. One hundred and thirteen women (71 per cent) out of 159 women followed through delivery were parasitaemic at least once. Mean placental haemozoin concentrations were significantly higher in women with delivery parasitaemias (223 ng/mg protein) than in women who never had a detectable parasitaemia (43 ng/mg protein; P<0.05), but were not significantly higher in women who were parasitaemic only during the antenatal period (67 ng/mg protein). Haemozoin was not associated with preterm delivery (PTD) or intrauterine growth retardation (IUGR) (P -values, 0.307-0.787). Thus, placental haemozoin is associated with malaria infection at the time of delivery and does not seem to be associated with poor birth outcome. PMID- 10833380 TI - Protection of murine gestational tissues from picornavirus infection in the preimplantation period. AB - Mice were inoculated with Theiler's murine encephalomyelitis virus (TMEV) on gestational days 1-3 (pre-implantation) or days 4-5 (peri- or post-implantation) or with control cell lysate (days 1-5). Dams were subsequently sacrificed between days 11-14 of gestation, and placentae and fetuses were harvested. Few placentae from dams inoculated with virus on days 1-3 were positive by virus culture (2 per cent) or in situ hybridization (6 per cent), and no fetuses were positive by either technique. In contrast, most placentae from dams inoculated with virus on days 4-5 were virus-positive by culture (96 per cent) or in situ hybridization (100 per cent), and a moderate number of fetuses were also positive (30 per cent by culture, 19 per cent by in situ hybridization). Necrosis was present more frequently in placentae from mice inoculated with virus on days 4-5 (55 per cent) than in placentae from dams inoculated with virus on days 1-3 (19 per cent) or with control cell lysate (18 per cent). Viral infection, mononuclear inflammation and cell necrosis were identified in the heart and great vessels of TMEV-infected fetuses. These results indicate that gestational tissues are largely protected from viral infection before implantation. After implantation, gestational tissues are more readily infected and damaged by maternal picornavirus infection. PMID- 10833381 TI - Adrenal tissue in the placenta: a heterotopia caused by migration and embolism? AB - Heterotopic adrenal tissue is not uncommon, especially in the urogenital system. Adrenocortical tissue in the placenta, however, is presumably very rare. To our knowledge, four cases have been published. There are several different theories to explain such a heterotopia. According to our findings, an embolic spread of adrenal precursor cells via fetal vascular shortcuts is the most likely mechanism. Apart from that hypothesis, the possibility of a monodermal teratoma as well as of an aberrant differentiation of cells of the extraembryonic mesoderm are considered in the literature. PMID- 10833382 TI - Umbilical cord ulcer: a serious in utero complication of intestinal atresia. AB - We have experienced six cases of umbilical cord ulcer that resulted in life threatening antenatal fetal haemorrhage. Fetal distress and bloody amniotic fluid that led to intrauterine and neonatal death in four patients complicated the pregnancy courses. The remaining two infants are still alive. All the infants had upper intestinal atresia below the papilla of Vater. Pathologically, umbilical arteries and surrounding Wharton's jelly were necrotic in all cases. Activated macrophages containing pigment granules were present within the ulcer bed, necrotic areas of Wharton's jelly and fetal membranes. During the same period, we encountered 38 other cases of upper intestinal atresia and the placentae were examined in 23 cases. Microscopic degeneration of Wharton's jelly was seen in three out of 23 cases. Total bile acids content in amniotic fluid was elevated in one case of umbilical cord ulcer. We speculate that the umbilical cord ulcer is closely related to in utero regurgitation of the bile. We also think that the incidence of this serious association is higher than has been reported previously. PMID- 10833383 TI - The 15-deoxy-delta(12,14)-prostaglandin J(2)receptor, peroxisome proliferator activated receptor-gamma (PPARgamma) is expressed in human gestational tissues and is functionally active in JEG3 choriocarcinoma cells. AB - RNA was extracted from human gestational membranes and villous placental tissue following spontaneous delivery (n = 15) or elective caesarean section (n = 15) at term. The samples were subjected to Northern analysis, using a 2 kb cDNA probe for peroxisome proliferator activated receptor (PPAR)-gamma. The mRNA was detectable in all choriodecidual and villous placental samples, irrespective of mode of delivery, but was only rarely detectable in the amnion. The JEG3 choriocarcinoma cell line also expressed PPARgamma. In order to evaluate PPAR mediated transcriptional activation in JEG3 cells, the cells were transfected with pTK-PPREx3-luc, a PPAR response element (PPRE) containing luciferase reporter construct. Subsequent treatment with 10 microm 15-deoxy delta(12,14)prostaglandin J(2)(15dPGJ(2)) resulted in an eight-fold stimulation of luciferase production relative to controls transfected with the same construct lacking the PPRE. This stimulation was concentration-dependent. These results suggest roles for PPARgamma and its ligand in lipid, steroid and inflammatory mediator homeostasis and in remodelling of gestational tissues. PMID- 10833384 TI - Overexpression and purification of human XPA using a baculovirus expression system. AB - The xeroderma pigmentosum group A protein (XPA) is an essential component of the eukaryotic nucleotide excision repair (NER) process. Recombinant human XPA was expressed in baculovirus-infected insect cells as a [His](6)-tagged fusion protein. A two-column purification procedure resulted in greater than 90% purity for the recombinant protein with a final yield of 0.53 mg from 200 ml of infected cells. The recombinant protein migrated as a doublet of 44 and 42 kDa upon SDS PAGE consistent with that observed for the native protein. XPA can interact with a number of proteins including replication protein A (RPA) which has been implicated in the initial recognition of damaged DNA. Using a modified ELISA, we demonstrate that the recombinant XPA fusion protein also forms a complex with RPA independent of DNA. The ability of XPA to bind damaged DNA was assessed in an electrophoretic mobility shift assay using globally cisplatin-damaged DNA. The results revealed a slight preference for DNA damaged with cisplatin consistent with its proposed role in the recognition of damaged DNA. The recombinant XPA fusion protein was able to complement cell-free extracts immunodepleted of XPA restoring NER-catalyzed incision of cisplatin-damaged DNA in an in vitro excision repair assay. PMID- 10833385 TI - Generation of a baculovirus recombinant prostate-specific membrane antigen and its use in the development of a novel protein biochip quantitative immunoassay. AB - Prostate-specific membrane antigen (PSMA) is a 100-kDa transmembrane glycoprotein identified by the monoclonal antibody 7E11-C5.3 from the human prostate tumor cell line LNCaP. Because of its significant upregulation in androgen refractory and metastatic prostate cancers, PSMA may be a useful prognostic biomarker and a target for developing novel therapeutic strategies. However, the lack of abundant pure PSMA protein and the low efficacy in immunoaffinity isolation from LNCaP cells have hampered the development of clinical assays. In order to obtain a renewable and reliable source of pure antigen, we used the baculovirus/insect cell system to express and purify a recombinant PSMA. A recombinant baculovirus containing a 6x histidine-tagged PSMA gene was generated, from which rPSMA was expressed and purified using cobalt-chelating affinity chromatography. The purity and correct molecular size of rPSMA were demonstrated by gel electrophoresis and mass spectrometry. Glycosidase digestions showed that the oligosaccharides on rPSMA are primarily N-linked high-mannose type. Although the glycosylation is different from the native PSMA, it did not affect the immunoreactivity of rPSMA to antibodies specific for either the intra- or the extracellular domains of PSMA. Finally, the purified rPSMA was successfully used to develop a quantitative PSMA immunoassay using the novel ProteinChip surface-enhanced laser desorption/ionization mass spectrometry technology. PMID- 10833386 TI - Expression and purification of recombinant human indoleamine 2, 3-dioxygenase. AB - Indoleamine 2,3-dioxygenase, the first and rate-limiting enzyme in human tryptophan metabolism, has been implicated in the pathogenesis of many diseases. The human enzyme was expressed in Escherichia coli EC538 (pREP4) as a fusion protein to a hexahistidyl tag and purified to homogeneity in terms of electrophoretic and mass spectroscopic analysis, by a combination of phosphocellulose and nickel-agarose affinity chromatography. The yield of the fusion protein was 1.4 mg per liter of bacterial culture with an overall recovery of 56% from the crude extract. When the culture medium was supplemented with 7 microM hemin, the purified protein contained 0.8 mol of heme per mole of enzyme and exhibited an absorption spectrum consistent with the ferric form of hemoprotein. The pI value of the recombinant enzyme was 7.09 compared with 6.9 for the native enzyme. This was as expected from the addition of the hexahistidyl tag. Similar to the native enzyme, the recombinant enzyme required methylene blue and ascorbic acid for enzyme activity and oxidized not only l-tryptophan but also d-tryptophan and 5-hydroxy-l-tryptophan. The molecular activities for these substrates and their K(m) values were similar to those of the native enzyme, indicating that the addition of the hexahistidyl tag did not significantly affect catalytic activity. The recombinant protein can therefore be used to investigate properties of the native enzyme. This will aid the development of specific inhibitors of indoleamine 2,3-dioxygenase, which may be effective in halting disease progression. PMID- 10833387 TI - Preparation of recombinant bovine, porcine, and porcine W4R/R5K leptins and comparison of their activity and immunoreactivity with ovine, chicken, and human leptins. AB - Recombinant ovine Ala-leptin (GenBank Accession No. U84247, of ovine leptin), previously prepared in our laboratory in prokaryotic expression plasmid pMON3401, was mutated using a mutagenesis kit to prepare plasmids encoding for bovine (GenBank Accession No. U50365) and porcine (GenBank Accession No. U59894) leptins and for porcine leptin analogue W4R/R5K. Escherichia coli cells transformed with these plasmids overexpressed large amounts of these proteins upon induction with nalidixic acid. The expressed proteins, found in inclusion bodies, were refolded and purified to homogeneity using subsequently anion- and cation-exchange chromatography. All three purified proteins showed a single band of the expected molecular mass of 16 kDa in SDS-PAGE in the presence of reducing agent and were composed of 90-100% monomers. Proper refolding was evidenced by comparing their CD spectra to those of previously prepared chicken and ovine leptins and to commercially available human leptin. The amino acid content of the purified proteins closely resembled the predicted composition. The biological activity of bovine leptin, porcine leptin, and porcine leptin analogue W4R/R5K was evidenced by their ability to stimulate proliferation of leptin-sensitive BAF/3 cells transfected with a long form of human leptin receptor. All three proteins, as well as ovine and chicken leptins, but not human leptin, exhibited a very high degree of cross-immunoreactivity against antiserum raised against ovine leptin in rabbits. In contrast, none or very low cross-immunoreactivity was observed against antiserum raised against ovine leptin in goats. PMID- 10833388 TI - Expression, refolding, and activity of a recombinant nonhemorrhagic snake venom metalloprotease. AB - Snake venoms are rich sources of proteases that strongly affect the vascular system, by promoting blood coagulation, hemorrhage, and fibrinolysis. Hemorrhagic activity is mostly due to the enzymatic action of metalloproteases on capillary basement membrane components, such as collagen IV, laminin, and fibronectin. A few low-molecular-weight snake venom metalloproteases (svMP) have been described as being devoid of hemorrhagic activity, but they have strong direct-acting fibrinolytic activity that could be very helpful in thrombosis therapy. We have developed an expression system for production of a recombinant svMP from a cDNA (ACLPREF) coding for a small metalloprotease (ACLF) with three disulfide bonds from an Agkistrodon contortrix laticinctus (broad-banded copperhead) venom gland cDNA library. The mature protein-coding region was amplified by PCR and subcloned into the pET28a vector, and the resulting plasmid was used to transform BL21(DE3) Escherichia coli cells. Culture of the transformants at either 37 or 20 degrees C led to the overexpression of an insoluble and inactive 30-kDa protein after 1.0 mM IPTG induction. The expressed protein (rACLF) was recovered from inclusion bodies with 6 M buffered urea solution and purified on a nickel-Sepharose column followed by gel filtration chromatography, both under denaturing conditions. After treatment with dithiothreitol, protein refolding was performed by gradual removal of the denaturing agent by dialysis. The refolded recombinant protein was active in fibrin-agarose plates. The purified protein achieved a conformation similar to that of the native enzyme as judged by circular dichroism analysis. PMID- 10833389 TI - Overexpression and purification of fructose-1-phosphate kinase from Escherichia coli: application to the assay of fructose 1-phosphate. AB - Fructose 1-phosphate is a metabolite that plays a regulatory role in liver and is best measured using an assay based on its conversion to fructose 1,6-bisphosphate by a bacterial fructose-1-phosphate kinase (Fru1PK). The open reading frame encoding Escherichia coli Fru1PK has been introduced in an expression plasmid (pET3a) based on the T7 promoter-driven system, which was used to overexpress the enzyme. The conditions for the production of soluble Fru1PK were optimized. The purification procedure used involved ammonium sulfate precipitation and chromatography on DEAE-Sepharose and was aimed mostly at stabilizing the enzyme and at freeing Fru1PK from bacterial contaminants that could interfere in the fructose 1-phosphate assay. From a 1-liter culture, more than 50 mg protein is obtained. This preparation can be used in an enzymatic assay that measures specifically fructose 1-phosphate in tissue extracts. PMID- 10833390 TI - The preparation of apo-Cu,Zn superoxide dismutase by ion-exchange chromatography on iminodiacetic acid-sepharose. AB - The superoxide dismutases (EC 1.15.1.1) are a family of enzymes that catalyze the dismutation of superoxide radical anion to dioxygen and hydrogen peroxide. The active site contains a critical metal ion such as manganese, iron, or copper. The copper-containing protein also has one zinc ion bound per subunit. The standard method used to remove the metal ions from Cu,Zn superoxide dismutase has been to exhaustively dialyze the protein against chelating agents at low pH. We have developed a new method where the protein is bound to ion-exchange medium based on iminodiacetic acid immobilized on Sepharose. The bound protein is treated with a buffer containing edta at pH 3.5 to remove metal ions; the buffer is then exchanged for acetate buffer to remove edta, after which the protein is eluted by a salt gradient. An advantage of this method is that a single chromatography step is sufficient to produce apo protein. Results are shown for both human and bovine dimeric Cu,Zn superoxide dismutase and the monomeric Escherichia coli Cu,Zn superoxide dismutase. In every case, the metals were removed efficiently. PMID- 10833391 TI - Purification of histidine-tagged mitochondrial ADP/ATP carrier: influence of the conformational states of the C-terminal region. AB - A functional recombinant mitochondrial ADP/ATP carrier from the yeast Saccharomyces cerevisiae that bears a six-histidine tag at the C-terminus, Anc2(His(6))p, has been engineered to allow its purification by immobilized metal ion affinity chromatography (IMAC). The tagged carrier was expressed at a level similar to that of unmodified Anc2p as determined by immunodetection and titration of the specific atractyloside binding sites. Anc2(His(6))p, enriched by chromatography on hydroxyapatite of detergent extracts of mitochondria, was still contaminated by mitochondrial proteins and a large amount of ergosterol. It was highly purified after adsorption on Ni-NTA resin and elution by imidazole buffer, with a 90-95% overall yield. Anc2(His(6))p interacted differently with immobilized ions depending on whether it was unliganded or bound to carboxyatractyloside (CATR) or bongkrekic acid (BA), two specific inhibitors of the ADP/ATP transport, thus indicating that accessibility of the C-terminus is markedly influenced by the conformational state of the carrier. Fluorometric assays demonstrated that purified unliganded Anc2(His(6))p was in a functional state since it underwent CATR- and BA-sensitive and ADP (or ATP)-induced conformational changes. Large-scale purification of Anc2(His(6))p-CATR and Anc2(His(6))p-BA complexes by IMAC will be of major interest for structural analysis of the ADP/ATP carrier. PMID- 10833392 TI - Expression and purification of soluble and inactive mutant forms of membrane type 1 matrix metalloproteinase. AB - Membrane type 1 matrix metalloproteinase (MT1-MMP) is a membrane-bound proteinase and a cell-surface receptor and activator of gelatinase A in normal and neoplastic cells. We have expressed and purified a soluble deletion mutant of MT1 MMP lacking the transmembrane and cytoplasmic domains and an inactive mutant of the soluble MT1-MMP, where the active-site glutamic acid(240) was substituted by alanine (E240A). A baculovirus transfer vector coding for amino acids 21-539 of MT1-MMP (DeltaTM) and a similar vector coding for the mutation (E240ADeltaTM) were constructed for expression in insect cells. Both DeltaTM and E240ADeltaTM were secreted to the culture medium of infected High Five insect cells. They were then purified by cation-exchange followed by gel-filtration chromatography. DeltaTM was able to cleave denatured type I collagen and fibronectin and activate MMP-2/gelatinase-A, while E240ADeltaTM had only low proteolytic activity against denatured collagen I. The current expression and purification protocol should prove useful for the production of large amounts of enzymatically active soluble MT1-MMP. PMID- 10833393 TI - Heterologous expression in Pseudomonas aeruginosa and purification of the 9.2-kDa c-type cytochrome subunit of p-cresol methylhydroxylase. AB - The 9.2-kDa c-type cytochrome subunit (PchC) of the flavocytochrome p-cresol methylhydroxylase from Pseudomonas putida NCIMB 9869 has been overexpressed in recombinant form in Pseudomonas aeruginosa PAO1-LAC, using the recently developed pUCP-Nde vector. Efforts to produce the cytochrome in Escherichia coli using a pET vector, with or without its signal peptide, were generally unsuccessful, yielding relatively low levels of the protein. In contrast, the mature form of PchC accumulated in the periplasmic space of P. aeruginosa PAO1-LAC to about 1 mg/g wet cell paste. A periplasmic fraction enriched to about 12% (w/w) of total protein with recombinant PchC was isolated from the remainder of the cells by a washing procedure using ethylenediaminetetraacetate in the presence of sucrose. The cytochrome was purified to homogeneity from the periplasmic extract by anion exchange chromatography on DEAE-Sepharose CL-6B followed by chromatofocusing on PolyBuffer Exchanger 94. Purified PchC was obtained in a yield of about 50% and was shown to be identical to that resolved from the native flavocytochrome isolated from P. putida. This system may prove to be of general use for the production of recombinant c-type cytochromes. PMID- 10833394 TI - Overexpression in Escherichia coli, purification, and characterization of Sphingomonas sp. A1 alginate lyases. AB - A bacterium Sphingomonas sp. A1 produces three kinds of alginate lyases [A1-I (66 kDa), A1-II (25 kDa), and A1-III (40 kDa)] from a single precursor, through posttranslational processing. Overexpression systems for these alginate lyases were constructed in Escherichia coli cells by controlling of the lyase genes under T7 promoter and terminator. Expression levels of A1-I, A1-II, and A1-III in E. coli cells were 3.50, 3.04, and 2.13 kU/liter of culture, respectively, and were over 10-fold higher than those in Sphingomonas sp. A1 cells. Purified A1-I, A1-II, and A1-III from E. coli cells were monomeric enzymes with molecular masses of 63, 25, and 40 kDa, respectively. The depolymerization pattern of alginate with A1-I and A1-II indicated that both enzymes cleaved the glycosidic bond of the polymer endolytically and by beta-elimination reaction. A1-II preferred polyguluronate rather than polymannuronate and released tri- and tetrasaccharides, which have unsaturated uronyl residues at the nonreducing terminal, from alginate as the major final products. A1-I acted equally on both homopolymers and produced di- and trisaccharides as the final products. PMID- 10833395 TI - Functional human insulin-degrading enzyme can be expressed in bacteria. AB - Insulin-degrading enzyme (IDE) has been shown to degrade a number of biologically important peptides, including insulin and the amyloid-beta protein implicated in Alzheimer's disease. However, lack of a facile method to generate purified enzyme and related mutants has made it difficult to study the precise role of IDE in the clearance of these peptides. Therefore, we determined whether recombinant wild type and mutant human IDEs can be overexpressed as functional enzymes in bacteria. Three vectors carrying cDNAs encoding N-terminally polyhistidine-tagged recombinant IDEs were constructed, and the proteins expressed in Escherichia coli were purified by metal affinity chromatography (final yield approximately 8 mg per liter of culture). The recombinant IDEs, like the endogenous mammalian enzyme, migrate with 110-kDa apparent molecular masses in SDS-polyacrylamide gels and as a approximately 200-kDa species in gel filtration. Further analysis by native PAGE indicates that IDE can form multimers of different complexities. The wild-type recombinant endopeptidase degrades insulin with an efficiency similar to that of the enzyme purified from mammalian tissues. Purified IDEs are stable at 4 degrees C for at least 1 month. Purified recombinant protein was used to raise specific polyclonal antibodies that can immunoprecipitate native mammalian IDE. Thus, the procedure described allows the rapid production of large amounts of purified IDE and demonstrates that IDE can be produced in an active form in the absence of other potential interacting mammalian proteins. PMID- 10833396 TI - Active recombinant human tyrosine kinase c-Yes: expression in baculovirus system, purification, comparison to c-Src, and inhibition by a c-Src inhibitor. AB - C-Yes is a non-receptor-type tyrosine kinase of the Src family that is most closely related to c-Src. C-Yes has been implicated in development of some human cancers. Here we report on the expression, purification, and characterization of the active human recombinant c-Yes. A full-length human c-Yes clone has been generated and the protein was expressed in insect Sf9 cells. Active c-Yes was purified by liquid chromatography to yield a preparation with a high specific activity (160 nmol/min/mg using an optimal Src substrate peptide). In a comparison between human c-Yes and c-Src enzymes, relative phosphorylation efficiencies on nine protein and four peptide substrates were different. However, the recently described Src inhibitor CGP77675 inhibited human c-Yes with a potency similar to that of c-Src (IC(50) value of 6.5 nM). The purified preparation of active c-Yes provides a good basis for further enzymatic characterization and for the development of c-Yes-specific inhibitors. PMID- 10833397 TI - Cloning, expression, and purification of the His(6)-tagged thermostable beta galactosidase from Pyrococcus woesei in Escherichia coli and some properties of the isolated enzyme. AB - In the previous study we cloned Pyrococcus woesei gene coding thermostable beta galactosidase into pET30-LIC expression plasmid. The nucleotide sequence revealed that beta-galactosidase of P. woesei consists of 510 amino acids and has a molecular weight of 59, 056 kDa (GenBank Accession No. AF043283). It shows 99.9% nucleotide identity to the nucleotide sequence of beta-galactosidase from Pyrococcus furiosus. We also demonstrated that thermostable beta-galactosidase can be produced with high yield by Escherichia coli strain and can be easy separated by thermal precipitation of other bacterial proteins at 85 degrees C (S. D $$;abrowski, J. Maciunska, and J. Synowiecki, 1998, Mol. Biotechnol. 10, 217-222). In this study we presented a new expression system for producing P. woesei beta-galactosidase in Escherichia coli and one-step chromatography purification procedure for obtaining pure enzyme (His(6)-tagged beta galactosidase). The recombinant beta-galactosidase contained a polyhistidine tag at the N-terminus (20 additional amino acids) that allowed single-step isolation by Ni affinity chromatography. The enzyme was purified by heat treatment (to denature E. coli proteins), followed by metal-affinity chromatography on Ni(2+) TED-Sepharose columns. The enzyme was characterized and displayed high activity and thermostability. This bacterial expression system appears to be a good method for production of the thermostable beta-galactosidase. PMID- 10833398 TI - Comparative characterization of two forms of recombinant human SPC1 secreted from Schneider 2 cells. AB - SPC1 (furin/PACE), an enzyme belonging to the S8 group of serine endoproteases, is a type I integral membrane protein that catalyzes the processing of a multitude of precursor proteins. We report here the use of transfected Drosophila melanogaster Schneider 2 cells to produce milligram amounts of two forms of recombinant human SPC1. In order to investigate the role of the cysteine-rich region (CRR) of SPC1, we compared the biochemical and enzymatic properties of hSPC1/714 that has the C-terminal tail and transmembrane region of the native enzyme removed with that of hSPC1/585 which had, in addition, the CRR deleted. Two stable cell lines were established. The S2-hSPC1/714 line secreted a major form of apparent molecular weight of 83 kDa and a minor form of 80 kDa whereas the S2-hSPC1/585 line secreted a single 59-kDa protein. PNGase F treatment of the different forms demonstrated that the enzymes were glycosylated. Automated NH(2) terminal sequencing revealed that all purified forms resulted from processing at the expected zymogen activation site. Removal of the CRR resulted in a broadening of the enzyme's pH range, a shift of K(0.5) for Ca(2+), and a shorter enzymatic half-life when compared to the longer form, which suggest that the CRR of hSPC1 may help in stabilizing the enzyme's proteolytic activity. The use of this high level expression system will meet the demand for material necessary to perform biochemical and structural studies that are needed to further our understanding of this and other SPCs at the molecular level. PMID- 10833399 TI - Cloning, expression, and one-step purification of the minimal essential domain of the light chain of botulinum neurotoxin type A. AB - A truncated but functional form of the botulinum neurotoxin A light chain (Tyr 9 Leu 415) has been cloned into the three bacterial expression vectors, pET 28, pET 30, and PGEX-2T, and produced as fusion proteins. This 406-amino-acid light chain was expressed with 1 six-histidine tag (LC-pET28), 2 six histidine tags and a S tag (LC-pET30), or a six-histidine tag and a glutathione S-transferase tag (LC pGEX-2T). The three fusion proteins have been overexpressed in Escherichia coli, purified in a soluble form, and tested for protease activity. All three recombinant proteins were found to have similar enzymatic activity, comparable to the light chain purified from the whole toxin. The LC-pET30 protein was the most soluble and stable of the three fusion proteins, and it could be purified using a one-step affinity chromatography protocol. The purified protein was determined to be 98% pure as assessed by SDS-polyacrylamide gel. This protein has been crystallized and initial X-ray data show that the crystals diffract to 1.8 A. PMID- 10833400 TI - Production and characterization of biologically active human GM-CSF secreted by genetically modified plant cells. AB - Human granulocyte-macrophage colony-stimulating factor (GM-CSF), a hemopoietic growth factor, was produced and secreted from tobacco cell suspensions. The GM CSF cDNA was carried by a binary vector under the control of the CaMV 35S promoter and the T7 terminator. In addition, a 5'-nontranslated region from the tobacco etch virus (TEV leader sequence) was fused to the N-terminal end of the GM-CSF transgene. For ease of purification, a 6-His tag was added to the 3' end of the GM-CSF cDNA. Addition of the TEV leader sequence increased protein production more than twofold compared to non-TEV controls. Initial batch cultivation studies indicated a maximum of 250 microg/L extracellular and 150 microg/L intracellular GM-CSF. Western blot analysis detected multiple peptides with masses from 14 to 30 kDa in the extracellular medium. The plant-produced GM CSF was biologically active and could be bound to a nickel affinity matrix, indicating that both the receptor-binding region and the 6-His tag were functional. The batch production of GM-CSF was compared with the production of other recombinant proteins secreted by transformed tobacco cells. The recovery of secreted GM-CSF was increased by the addition of stabilizing proteins and by increasing salt in the growth medium to physiological levels. PMID- 10833401 TI - Yeast cytochrome c peroxidase expression in Escherichia coli and rapid isolation of various highly pure holoenzymes. AB - A more efficient 2-day isolation and purification method for recombinant yeast cytochrome c peroxidase produced in Escherichia coli is presented. Two types of recombinant "wild-type" CcP have been produced and characterized, the recombinant nuclear gene sequence and the 294-amino-acid original protein sequence. These two sequences constitute the majority of the recombinant "native" or wild-type CcP currently in production and from which all recombinant variants now derive. The enzymes have been subjected to extensive physical characterizations, including sequencing, UV-visible spectroscopy, HPLC, gel electrophoresis, kinetic measurements, NMR spectroscopy, and mass spectrometry. Less extensive characterization data are also presented for recombinant, perdeuterated CcP, an enzyme produced in >95% deuterated medium. All of these results indicate that the purified recombinant wild-type enzymes are functionally and spectroscopically identical to the native, yeast-isolated wild-type enzyme. This improved method uses standard chromatography to produce highly purified holoenzyme in a more efficient manner than previously achieved. Two methods for assembling the holoenzyme are described. In one, exogenous heme is added at lysis, while in the other heme biosynthesis is stimulated in E. coli. A primary reason for developing this method has been the need to minimize loss of precious, isotope-labeled enzyme and, so, this method has also been used to produce both the perdeuterated and the (15)N-labeled enzyme, as well as several variants. PMID- 10833402 TI - Folding and structural characterization of highly disulfide-bonded beetle antifreeze protein produced in bacteria. AB - The hyperactive antifreeze protein from the beetle, Tenebrio molitor, is an 8.5 kDa, threonine-rich protein containing 16 Cys residues, all of which are involved in disulfide bonds. When produced by Escherichia coli, the protein accumulated in the supernatant in an inactive, unfolded state. Its correct folding required days or weeks of oxidation at 22 or 4 degrees C, respectively, and its purification included the removal of imperfectly folded forms by reversed-phase HPLC. NMR spectroscopy was used to assess the degree of folding of each preparation. One dimensional (1)H and two-dimensional (1)H total correlation spectroscopy spectra were particularly helpful in establishing the characteristics of the fully folded antifreeze in comparison to less well-folded forms. The recombinant antifreeze had no free -SH groups and was rapidly and completely inactivated by 10 mM DTT. It had a thermal hysteresis activity of 2.5 degrees C at a concentration of 1 mg/ml, whereas fish antifreeze proteins typically show a thermal hysteresis of approximately 1.0 degrees C at 10-20 mg/ml. The circular dichroism spectra of the beetle antifreeze had a superficial resemblance to those of alpha-helical proteins, but deconvolution of the spectra indicated the absence of alpha-helix and the presence of beta-structure and coil. NMR analysis and secondary structure predictions agree with the CD data and are consistent with a beta-helix model proposed for the antifreeze on the basis of its 12-amino-acid repeating structure and presumptive disulfide bond arrangement. PMID- 10833403 TI - Expression of active monomeric and dimeric nuclease A from the gram-positive Streptococcus gordonii surface protein expression system. AB - We used the surface protein expression (SPEX) system to express an anchored and a secreted form of staphylococcal nuclease A (NucA) from gram-positive bacteria. NucA is a small ( approximately 18 kDa), extracellular, monomeric enzyme from Staphylococcus aureus. A deletion of amino acids 114-119 causes monomeric NucA to form homodimers. The DNA sequence encoding either wild-type or deletion mutant NucA was cloned via homologous recombination into Streptococcus gordonii. S. gordonii strains expressing either anchored or secreted, monomeric or dimeric NucA were isolated and tested for enzymatic activity using a novel fluorescence enzyme assay. We show that active monomeric and dimeric NucA enzyme can be expressed either anchored on the cell surface or secreted into the culture medium. The activity of the dimer NucA was approximately 100-fold less than the monomer. Secreted and anchored, monomeric NucA migrated on SDS-polyacrylamide gels at approximately 18 or approximately 30 kDa, respectively. In addition, similar to S. aureus NucA, the S. gordonii recombinant NucA enzyme was dependent on CaCl(2) and was heat stable. In contrast, however, the recombinant NucA activity was maximal at pH 7.0-7.5 whereas S. aureus NucA was maximal at pH 9.0. These results show, for the first time, expression of active enzyme and polymeric protein in secreted and anchored forms using SPEX. This further demonstrates the utility of this gram-positive surface protein expression system as a potential commensal bacterial delivery system for active, therapeutic enzymes, biopharmaceuticals, or vaccines. PMID- 10833404 TI - High-level expression in Escherichia coli and purification of the membrane-bound form of cytochrome b(5). AB - Expression of the membrane-bound form of rabbit cytochrome b(5) in Escherichia coli has been significantly improved through the use of the T7 expression vector pLW01 (A. Bridges, L. Gruenke, Y.-T. Chang, I. Vakser, G. Loew, and L. Waskell, 1998, J. Biol. Chem. 273, 17036-17049) in conjunction with strain C41(DE3) (B. Miroux and J. Walker, 1996, J. Mol. Biol. 260, 289-298). Cell cultures expressing the cytochrome b(5) contained an average of 820 mg/liter of culture and reached peak levels as high as 1100 mg/liter when higher antibiotic concentrations were used. Maximal levels were obtained from cultures when expression was induced with 10 microM IPTG. Approximately 90% of the cytochrome b(5) was expressed as apoprotein which was reconstituted by addition of exogenous heme. The cytochrome b(5) was purified from detergent-solubilized bacterial membranes using anion exchange chromatography on DEAE-Sepharose followed by size-exclusion chromatography on Superdex-75. Purification of cytochrome b(5) from a 500-ml culture yielded 121 mg of protein which had a specific content of 50 nmol of heme per milligram of protein with an overall recovery of 35%. The final cytochrome b(5) was free of any detectable contaminants when analyzed by SDS-PAGE. PMID- 10833405 TI - High-level production of recombinant fungal endo-beta-1,4-xylanase in the methylotrophic yeast Pichia pastoris. AB - Efficient production of recombinant Aspergillus niger family 11 1, 4-beta xylanase was achieved in Pichia pastoris. The cDNA-encoding XylA fused to the Saccharomyces cerevisiae invertase signal peptide was placed under the control of the P. pastoris AOX1 promoter. Secretion yields up to 60 mg/liter were obtained in synthetic medium. The recombinant XylA was purified to homogeneity using a one step purification protocol and found to be identical to the enzyme overexpressed in A. niger with respect to size, pI, and immunoreactivity. N-terminal sequence analysis of the recombinant protein indicated that the S. cerevisiae signal peptide was correctly processed in P. pastoris. The purified protein has a molecular weight of 19,893 Da, in excellent agreement with the calculated mass, and appears as one single band on isoelectric focusing with pI value around 3.5. Electrospray ionization mass spectrometry confirmed the presence of one major isoform produced by P. pastoris and the absence of glycosylation. The recombinant enzyme was further characterized in terms of specific activity, pH profile, kinetic parameters, and thermostability toward birchwood xylan as substrate and compared with the xylanase purified from A. niger. Both enzymes exhibit a pH optimum at 3.5 and maximal activity at 50 degrees C. The enzyme activity follows normal Michaelis-Menten kinetics with K(m) and V(max) values similar for both enzymes. P. pastoris produced recombinant xylanase in high yields that can be obtained readily as a single form. A. niger xylanase is the first microbial xylanase efficiently secreted and correctly processed by P. pastoris. PMID- 10833406 TI - Overexpression, purification, and crystallization of the membrane-bound fumarate reductase from Escherichia coli. AB - Quinol-fumarate reductase (QFR) from Escherichia coli is a membrane-bound four subunit respiratory protein that shares many physical and catalytic properties with succinate-quinone oxidoreductase (EC 1.3.99.1) commonly referred to as Complex II. The E. coli QFR has been overexpressed using plasmid vectors so that more than 50% of the cytoplasmic membrane fraction is composed of the four subunit enzyme complex. The growth characteristics required for optimal levels of expression with minimal degradation by host cell proteases and oxidation factors were determined for the strains harboring the recombinant plasmid. The enzyme is extracted from the enriched membrane fraction using the nonionic detergent Thesit (polyoxyethylene(9)dodecyl ether) in a monodisperse form and then purified by a combination of anion-exchange, perfusion, and gel filtration chromatography. The purified enzyme is highly active and contains all types of redox cofactors expected to be associated with the enzyme. Crystallization screening of the purified QFR by vapor diffusion resulted in the formation of crystals within 24 h using a sodium citrate buffer and polyethylene glycol precipitant. The crystals contain the complete four-subunit QFR complex, diffract to 3.3 A resolution, and were found to be in space group P2(1)2(1)2(1) with unit cell dimensions a = 96.6 A, b = 138.1 A, and c = 275.3 A. The purification and crystallization procedures are highly reproducible and the general procedure may prove useful for Complex IIs from other sources. PMID- 10833407 TI - Endogenous glutathione-binding proteins of insect cell lines: characterization and removal from glutathione S-transferase (GST) fusion proteins. AB - After affinity purification on immobilized glutathione, insect-cell-derived glutathione S-transferase (GST) fusion proteins contain variable amounts of protein contaminants of about 23-24 kDa. We have isolated these glutathione binding proteins from the widely used Sf9 and Hi5 insect cell lines and characterized them by LC-MS and N-terminal sequencing. Based on the observation that these proteins have higher affinity for glutathione than GST fusions, we have found that by using differential elution conditions the amount of such contaminants in GST fusion preparations can be strongly reduced directly during the affinity purification step. The main interest of these results is that they are not restricted to a specific construct, but rather they seem to apply to various insect-cell-derived GST fusions. PMID- 10833408 TI - Purification and characterization of recombinant human alpha-N acetylglucosaminidase secreted by Chinese hamster ovary cells. AB - alpha-N-Acetylglucosaminidase (EC 3.2.1.50) is a lysosomal enzyme that is deficient in the genetic disorder Sanfilippo syndrome type B. To study the human enzyme, we expressed its cDNA in Lec1 mutant Chinese hamster ovary (CHO) cells, which do not synthesize complex oligosaccharides. The enzyme was purified to apparent homogeneity from culture medium by chromatography on concanavalin A Sepharose, Poros 20-heparin, and aminooctyl-agarose. The purified enzyme migrated as a single band of 83 kDa on SDS-PAGE and as two peaks corresponding to monomeric and dimeric forms on Sephacryl-300. It had an apparent K(m) of 0.22 mM toward 4-methylumbelliferyl-alpha-N-acetylglucosaminide and was competitively inhibited by two potential transition analogs, 2-acetamido-1,2-dideoxynojirimycin (K(i) = 0.45 microM) and 6-acetamido-6-deoxycastanospermine (K(i) = 0.087 microM). Activity was also inhibited by mercurials but not by N-ethylmaleimide or iodoacetamide, suggesting the presence of essential sulfhydryl residues that are buried. The purified enzyme preparation corrected the abnormal [(35)S]glycosaminoglycan catabolism of Sanfilippo B fibroblasts in a mannose 6 phosphate-inhibitable manner, but its effectiveness was surprisingly low. Metabolic labeling experiments showed that the recombinant alpha-N acetylglucosaminidase secreted by CHO cells had only a trace of mannose 6 phosphate, probably derived from contaminating endogenous CHO enzyme. This contrasts with the presence of mannose 6-phosphate on naturally occurring alpha-N acetylglucosaminidase secreted by diploid human fibroblasts and on recombinant human alpha-l-iduronidase secreted by the same CHO cells. Thus contrary to current belief, overexpressing CHO cells do not necessarily secrete recombinant lysosomal enzyme with the mannose 6-phosphate-targeting signal; this finding has implications for the preparation of such enzymes for therapeutic purposes. PMID- 10833409 TI - Functional and immunological analysis of recombinant mouse H- and L-ferritins from Escherichia coli. AB - The production and characterization of recombinant mouse H- and L-ferritin chains from Escherichia coli are described. The proteins were efficiently expressed and purified with yields of 7-40 mg per liter of cell culture. They had the expected molecular mass and showed a physical stability analogous to that of the corresponding human ferritins. Mouse H- and L-ferritins had a very similar mobility on denaturing SDS-PAGE, but could be readily separated on nondenaturing PAGE because of the distinct slow mobility of mouse L-ferritin. Direct comparative experiments showed that mouse and human H-ferritins had the same iron incorporation activity, whereas mouse L-ferritin incorporated iron less efficiently than human L-ferritin. The difference was attributed to the substitution of a residue exposed on the cavity surface (Glu140 --> Lys) in mouse L-ferritin, a hypothesis confirmed by the finding that the mouse L-ferritin mutant Lys140-Glu incorporated iron as efficiently as human L-ferritin. Rabbit antisera elicited by the recombinant mouse ferritins were specific for the H- and L-chains and did not cross-react with the human ferritins. The antibodies and the derived specific ELISA assays allow the determination of H- and L-ferritins in mouse tissues. PMID- 10833410 TI - The importance of adenosine deaminase for lymphocyte development and function. AB - Deficiency in the enzyme adenosine deaminase (ADA) in humans manifests primarily as severe lymphopenia and immunodeficiency, resulting in death by 6 months of age, if untreated. In this review, we discuss phenotypical, biochemical, and metabolic hallmarks of the disease, and describe a mouse model in which levels of ADA can be biochemically and genetically manipulated. This model provides exciting possibilities for uncovering the mechanisms by which this purine catabolic enzyme affects lymphopoiesis. PMID- 10833411 TI - Direct photoaffinity labeling of Kir6.2 by [gamma-(32)P]ATP-[gamma]4 azidoanilide. AB - ATP-sensitive potassium (K(ATP)) channels are under complex regulation by intracellular ATP and ADP. The potentiatory effect of MgADP is conferred by the sulfonylurea receptor subunit of the channel, SUR, whereas the inhibitory effect of ATP appears to be mediated via the pore-forming subunit, Kir6.2. We have previously reported that Kir6.2 can be directly labeled by 8-azido-[gamma (32)P]ATP. However, the binding affinity of 8-azido-ATP to Kir6.2 was low probably due to modification at 8' position of adenine. Here we demonstrate that Kir6.2 can be directly photoaffinity labeled with higher affinity by [gamma (32)P]ATP-[gamma]4-azidoanilide ([gamma-(32)P]ATP-AA), containing an unmodified adenine ring. Photoaffinity labeling of Kir6.2 by [gamma-(32)P]ATP-AA is not affected by the presence of Mg(2+), consistent with Mg(2+)-independent ATP inhibition of K(ATP) channels. Interestingly, SUR1, which can be strongly and specifically photoaffinity labeled by 8-azido-ATP, was not photoaffinity labeled by ATP-AA. These results identify key differences in the structure of the nucleotide binding sites on SUR1 and Kir6.2. PMID- 10833412 TI - A novel intracellular membrane-bound calcium-independent phospholipase A(2). AB - We have cloned human cDNA encoding a novel protein of 782 amino acids that contains the lipase consensus sequence Gly-Xaa-Ser-Xaa-Gly and several stretches surrounding the motif, which are homologous to those of the catalytic domain of cytosolic calcium-independent phospholipase A(2) (iPLA(2)). When expressed in COS 7 cells, the protein predominantly exists in the membrane fraction and exhibits a phospholipase A(2) activity in a calcium-independent manner. The transcript of the membrane-bound iPLA(2) gene is ubiquitously observed as a single band of approximately 3.3 kb on Northern blot, with the most abundant expression in the skeletal muscle and heart. By a search of the database, we have also identified its putative C. elegans homologue, which shows 47% identity with that of human in the iPLA(2) catalytic region. Thus the novel type of iPLA(2) is evolutionarily well conserved, suggestive of its biological significance. PMID- 10833413 TI - Adenosine-derived non-phosphate antagonists for P2Y(1) purinoceptors. AB - Novel type antagonists for P2Y(1) adenine nucleotide receptors were synthesized by coupling of adenosine 5'-OH group with oligo-aspartate chain via a carbonyl linker. All these conjugates (AdoOC(O)Asp(n), n = 1-4) inhibited the 2MeSADP stimulated synthesis of inositol phosphates in 1321N1 human astrocytoma cells stably expressing human P2Y(1) receptors. This inhibitory effect followed the rank order AdoOC(O)Asp(2)> AdoOC(O)Asp(3)> AdoOC(O)Asp(1)> AdoOC(O)Asp(4) with antagonistic constant pA(2) = 5.4 for AdoOC(O)Asp(2). Potency of this non phosphate inhibitor was comparable with the previously known adenosine 3',5'- and 2', 5'-bisphosphates. Chemical and biological stabilities of these novel adenosine derived antagonists of the nucleotide receptor provide perspectives of their pharmacological implication. PMID- 10833414 TI - Deencryption of cellular tissue factor is independent of its cytoplasmic domain. AB - Tissue factor (TF) is a transmembrane molecule that, when exposed to plasma, is the key initiator of coagulation. Cellular TF activity is normally "encrypted", but treating cells with calcium ionophore (i.e. , ionomycin or A23187) increases ("deencrypts") TF activity without increasing TF mRNA or antigen expression. Deencryption results from both plasma membrane phosphatidylserine (PS)-dependent and -independent mechanisms; however, the nature of the PS-independent component is unclear. Since deencryption has been suggested to result from release of TF dimers on the cell surface, and since TF's cytoplasmic domain binds to actin binding protein 280 and interacts with the cytoskeleton, we hypothesized that interactions with the cytoskeleton, through the cytoplasmic domain, play a role in mediating encryption/deencryption. We examined TF deencryption and the role of the cytoplasmic domain in the PS-independent component using baby hamster kidney (BHK) cells expressing full length TF (BHK-TF) or TF lacking its cytoplasmic domain (BHK-descyt) (Sorensen et al. (1999) J. Biol. Chem. 274, 21349). Both BHK TF and BHK-descyt cells exhibited a dose-dependent, 1.5- to 10-fold increase in TF activity upon treatment with calcium ionophore, and this increase in activity was only partially blocked by annexin V. These results indicate that deencryption is not restricted to cells which naturally express TF and that the PS-independent component of deencryption is intact on cells transfected with either full length or truncated TF. Our results clearly indicate that deencryption is not dependent on an intact cytoplasmic domain in transfected BHK cells. PMID- 10833415 TI - Molecular evidence of complex tissue- and sex-specific mRNA expression of the rat alpha(2u)-globulin multigene family. AB - alpha(2u)-Globulin is well known to be a rat protein encoded by a highly homologous multigene family with more than twenty members. We report here the cloning and identification of major alpha(2u)-globulin mRNA species expressed in various tissues. Initially, eight individual clones (PGCL1-8) were obtained from a male preputial gland cDNA library. Data base analysis with BLAST demonstrated six mRNAs to be novel, all clones being characterized by highly conserved sequence motifs as lipocalins. All cDNAs contained an open reading frame of 543 nucleotides and encode 181 amino acid proteins showing 92.5-98.7% and 87.3-98.3% nucleic and amino acid identity, respectively. Denaturing gradient gel electrophoresis (DGGE) with sequence analysis showed that PGCL4 is a major member in the female mammary gland, and in the submaxillary and lachrymal glands of both sexes, while the counterpart in male liver and the coagulate glands was found to be PGCL1. Numbers of cDNA species including PGCL1 and PGCL4 were found in preputial glands, no sex-related difference being observed. These results directly demonstrate complex tissue- and sex-specific expression of alpha(2u) globulins in terms of mRNA species, providing useful information for understanding regulation of the alpha(2u)-globulin multigene family. PMID- 10833416 TI - Activation of different lipoxygenase isozymes induces apoptosis in human erythroleukemia and neuroblastoma cells. AB - We investigated the ability of different hydroperoxides generated by lipoxygenase isozymes to induce programmed cell death (PCD) in human cells. Erythroleukemia K562 and neuroblastoma CHP100 cells were used, because they showed high basal activity of lipoxygenase. The hydroperoxides generated by 5-, 12-, or 15 lipoxygenases from linoleate, linolenate, or arachidonate, and the corresponding hydroxides, were able to induce PCD in both cell types, in a concentration- and time-dependent manner. After 24 h, K562 and CHP100 cells showed 2.5- to 3.5-fold more apoptotic bodies than the untreated controls. PCD elicited by lipoxygenase products was independent of intracellular glutathione concentration, and did not require mRNA transcription or protein synthesis. On the other hand, lipoxygenase products evoked an immediate and sustained rise in cytoplasmic calcium (within seconds), followed by mitochondrial uncoupling (within hours). Unlike the hydro(pero)xides, the terminal products of the arachidonate cascade (i.e., leukotrienes, prostaglandins and thromboxane) were not cytotoxic. PMID- 10833417 TI - Cyclic zinc-dithiocarbamate-S,S'-dioxide blocks CXCR4-mediated HIV-1 infection. AB - To test the anti-human immunodeficiency virus type-1 (HIV-1) activity of 3,6,9,12 tetraazatetradecane-1,14-diylbis(zinc dithiocarbamate)-S,S'-dioxide (cyclic zinc dithiocarbamate-S, S'-dioxide), MAGI and MAGIC-5 cells were used; the former express CXCR4 and the latter express both CXCR4 and CCR5, which are HIV-1 coreceptors. The compound markedly inhibited HIV-1 X4 (CXCR4-using) viral replication in both MAGI and MAGIC-5 cells. On the other hand, the replication of HIV-1 R5X4 (both CXCR4-and CCR5-using) in MAGI cells but not MAGIC-5 cells was inhibited by the compound. The compound was found to specifically inhibit HIV-1 (X4) envelope-mediated cell-to-cell fusion, binding of anti-CXCR4 monoclonal antibody (12G5) to CXCR4 expressed on the surface of cells, and calcium flux induced by stromal-derived factor-1alpha (SDF-1alpha) bound to CXCR4. The results suggest that the compound inhibited CXCR4-mediated HIV-1 infection by influencing to the HIV-1 coreceptor activity of CXCR4. PMID- 10833418 TI - Transforming growth factor-beta(1) increases the expression of lectin-like oxidized low-density lipoprotein receptor-1. AB - Lectin-like oxidized low-density lipoprotein (Ox-LDL) receptor-1 (LOX-1) is a novel cell-surface receptor for Ox-LDL, which can be expressed by vascular endothelial cells, smooth muscle cells, and macrophages. On the other hand, transforming growth factor (TGF)-beta(1), which plays crucial roles in vascular remodeling and the pathogenesis of atherosclerosis, has been shown to inhibit expression of class A scavenger receptors and CD36 in macrophages. Here we provide the evidence that TGF-beta(1) (0.1-10 ng/mL) induces LOX-1 protein and mRNA expression in both bovine aortic endothelial cells and smooth muscle cells in a dose- and time-dependent fashion, probably at the transcriptional level. TGF beta(1) also upregulates LOX-1 mRNA expression in murine peritoneal macrophages. Thus TGF-beta(1) can highly induce LOX-1 expression in vascular endothelial cells, smooth muscle cells, and macrophages, suggesting that TGF-beta(1) appears one of the key regulators that modulates expression of scavenger receptors. PMID- 10833419 TI - Transplantation of normal and DMD myoblasts expressing the telomerase gene in SCID mice. AB - The limited proliferative capacity of dystrophic human myoblasts severely limits their ability to be genetically modified and used for myoblast transplantation. The forced expression of the catalytic subunit of telomerase can prevent telomere erosion and can immortalize different cell types. We thus tested the ability of telomerase to immortalize myoblasts and analyzed the effect of telomerase expression on the success of myoblast transplantation. Telomerase expression did not significantly extend the human myoblast life span. The telomerase expressing myoblasts were nonetheless competent to participate in myofiber formation after infection with the retroviral vector. Although the new fibers obtained are less numerous than after the transplantation of normal myoblasts, these results demonstrate that the forced expression of telomerase does not block the ability of normal or dystrophic myoblasts to differentiate in vivo. It will be now necessary to determine the factors that prevent telomerase from extending the life span of human myoblasts before the potential of this intervention can be fully examined. PMID- 10833420 TI - A novel cell adhesion inhibitor, K-7174, reduces the endothelial VCAM-1 induction by inflammatory cytokines, acting through the regulation of GATA. AB - A novel inhibitor for the adhesion of monocytes to cytokine-stimulated endothelial cells, K-7174, was selected by an assay system using the cultured human monocytic cells and human endothelial cells. K-7174 inhibited the expression of vascular cell adhesion molecule-1 (VCAM-1) induced by either tumor necrosis factor alpha or interleukin-1beta, without affecting the induction of intercellular adhesion molecule-1 or E-selectin. K-7174 had no effect on the stability of VCAM-1 mRNA. Electrophoretic mobility shift assay revealed that its inhibitory effect on VCAM-1 induction was mediated by an effect on the binding to the GATA motifs in the VCAM-1 gene promoter region. K-7174 did not influence the binding to any of the following binding motifs: octamer binding protein, AP-1, SP 1, ets, NFkappaB, or interferon regulatory factor. These results suggest that the regulation of GATA binding may become a new target for anti-inflammatory drug development, acting through a mechanism independent from NFkappaB activity. PMID- 10833421 TI - p300/CBP-dependent and -independent transcriptional interference between NF kappaB RelA and p53. AB - p53 and NF-kappaB RelA are activated by various genotoxic agents and mutually suppress each other's ability to activate transcription, most likely through competition for transcriptional coactivators such as CBP or p300. However, we found that the inhibition by RelA of p53 transcriptional activity is not completely restored by CBP/p300 overexpression and that a p53 mutant can not suppress RelA activity despite of its ability to bind CBP/p300. In the present study, we further present evidence that these two transcriptional factors directly interact both in vivo and in vitro. These results therefore indicate that the cross transcriptional interference between p53 and RelA is partly caused by the direct interaction between these two transcription factors which is mediated by their dimerization/tetramerization domains and results in inhibition of each other's transcriptional activity. Finally, cells derived from RelA knockout mice showed enhanced p53 transcriptional activity, suggesting that this cross transcriptional interference is physiologically important in cellular response to genotoxic stress. PMID- 10833422 TI - PPAR-gamma is selectively upregulated in Caco-2 cells by butyrate. AB - The peroxisome proliferator-activated receptor (PPAR)-gamma is a nuclear lipid activable receptor controlling the expression of genes involved in lipid metabolism and adipocyte differentiation. In order to investigate the possible role of PPAR-gamma in the differentiation of intestinal epithelial cells, we examined its expression in the human colon carcinoma cell line Caco-2, which undergoes rapid cell differentiation in the presence of butyrate. PPARs were quantified on mRNA level by RT competitive multiplex PCR, the corresponding proteins were determined by Western blot. In contrast to PPAR-alpha and PPAR beta, PPAR-gamma mRNA and protein increased significantly in butyrate-treated Caco-2 cells in a dose- and time-dependent manner. This effect was butyrate specific, since no change in PPAR-gamma expression could be observed after incubation with propionate or valerate. Activation of PPAR-gamma by ciglitazone further increased butyrate-induced cell differentiation dose-dependently. These data demonstrate a role for PPAR-gamma in the regulation of cell differentiation in Caco-2 cells. PMID- 10833423 TI - Arp2/3 complex-independent actin regulatory function of WAVE. AB - We report that WAVE1/Scar1, a WASP-family protein that functions downstream of Rac in membrane ruffling, can induce part of the reorganization of the actin cytoskeleton without Arp2/3 complex. WAVE1 has been reported to associate and activate Arp2/3 complex at its C-terminal region that is rich in acidic residues. The deletion of the acidic residues abolished the interaction with and the activation ability of Arp2/3 complex. The expression of the mutant WAVE1 lacking the acidic residues (DeltaA), however, induced actin-clustering in cells as the wild-type WAVE1 did. In addition, this actin-clustering could not be suppressed by the coexpression of the Arp2/3 complex-sequestering fragment (CA-region) derived from N-WASP, which clearly inhibits Rac-induced membrane ruffling. This study therefore demonstrates that WAVE1 reorganizes the actin cytoskeleton not only through Arp2/3 complex but also through another unidentified mechanism that may be important but has been neglected thus far. PMID- 10833424 TI - A neuronal NO synthase (NOS1) gene polymorphism is associated with asthma. AB - Recent family-based studies have revealed evidence for linkage of chromosomal region 12q to both asthma and high total serum immunoglobulin E (IgE) levels. Among the candidate genes in this region for asthma is neuronal nitric oxide synthase (NOS1). We sought a genetic association between a polymorphism in the NOS1 gene and the diagnosis of asthma, using a case-control design. Frequencies for allele 17 and 18 of a CA repeat in exon 29 of the NOS1 gene were significantly different between 490 asthmatic and 350 control subjects. Allele 17 was more common in the asthmatics (0.83 vs 0.76, or 1.49 [95% CI 1.17-1.90], P = 0.013) while allele 18 was less common in the asthmatics (0.06 vs 0.12, or 0.49 [95% CI 0.34-0. 69], P = 0.0004). To confirm these results we genotyped an additional 1131 control subjects and found the frequencies of alleles 17 and 18 to be virtually identical to those ascertained in our original control subjects. Total serum IgE was not associated with any allele of the polymorphism. These findings provide support, from case-control association analysis, for NOS1 as a candidate gene for asthma. PMID- 10833425 TI - CD95/CD95 ligand-mediated counterattack does not block T cell cytotoxicity. AB - Expression of CD95 ligand on parenchymal, epithelial, or tumor cells has been suggested to downregulate the immune response and to control lymphocyte activation. Suppression might be mediated by induction of apoptosis or by inhibition of Ca(2+) channels upon CD95 triggering. We, therefore, aimed to employ this model to modify the immune response to an antigen presented to cytotoxic T cells by antigen-presenting MC57 cells. This model would be very useful to specifically downregulate the immune response to autoantigens in autoimmune situations. However, cytotoxic T cell lines tested in the present study were resistant to CD95 ligand expression on antigen-presenting MC57 cells. In addition, coincubation of the lymphocytes with antigen presenting cells failed to block cytotoxicity mediated by the T lymphocytes. We, therefore, conclude that single expression of CD95 ligand on antigen-presenting cells is insufficient to specifically downregulate an immune response by CD8(+-)triggered immune response. PMID- 10833426 TI - Identification of six CAG repeat domains into the human chromosomic region 12q24.1. AB - Six different domains of CAG repeats from a human chromosome 12-specific cosmid library were identified, cloned, and sequenced. These CAG repeat domains were localized into the human chromosomic region 12q24.1. Five of them constitute repeat candidates for expansions in autosomal dominant neurological disorders with genetic anticipation, and they can also contribute to the chromosome walking in the human genome project. PMID- 10833427 TI - Functional expression of the pore forming subunit of the ATP-sensitive potassium channel in Saccharomyces cerevisiae. AB - We have expressed the pore-forming subunits (Kir 6.1 and Kir 6.2) of the mammalian ATP-sensitive potassium channel in a potassium-transport deficient yeast strain (trk1 trk2). Functional expression of Kir 6.2 and Kir 6.1 can complement growth deficiency weakly and strongly respectively of the yeast strain on low-potassium medium. Mutations of Kir 6.2 that abolish ATP sensitivity (K185Q, I182Q) and enhance trafficking to the plasma membrane surface (Kir 6.2DeltaC36) lead to significantly better growth rescue. Growth rescue of Kir 6.1, Kir 6.2 and the above mutants can be inhibited by pharmacological agents (cesium ions, phentolamine and quinine) known to decrease channel activity by direct interaction with the pore forming subunit. Thus we have developed a system in yeast that can report both loss and gain of function mutations in these subunits and pharmacological interventions. PMID- 10833428 TI - Highly regulated expression of subtilisin-like proprotein convertase PACE4 (SPC4) during dentinogenesis. AB - Expressions of mRNAs for four subtilisin-like proprotein convertases (SPCs: furin, PACE4, PC6, and PC8) and bone morphogenetic protein 4 (BMP4) in the rat molar tooth during development were analyzed by Northern blotting, reverse transcriptase-polymerase chain reaction (RT-PCR), and in situ hybridization to explore the possible involvement of SPCs in the processing of proBMPs. We found a temporospacial expression of PACE4, but not one of the other SPCs, in this tissue; i.e., RT-PCR analysis revealed that the level of PACE4 mRNA, but not that of the other SPC mRNAs became high around the second postnatal day. This increase was in good accordance with the increase in BMP4 mRNA, indicating an apparent association of these molecules with the differentiation and establishment of functional ameloblasts and odontoblasts. During dentinogenesis, PACE4 mRNA was localized in the ameloblasts and odontoblasts. These observations suggest that PACE4 plays a crucial role in dentinogenesis, especially via the activation of BMPs. PMID- 10833429 TI - Mitochondrial GPx1 decreases induced but not basal oxidative damage to mtDNA in T47D cells. AB - The production of oxyradicals by mitochondria (mt) is a source of oxidative damage to mtDNA such as 8-oxo-dG lesions that may lead to mutations and mitochondrial dysfunction. The potential protection of mtDNA by glutathione peroxidase-1 (GPx1) was investigated in GPx1-proficient (GPx-2) and GPx1 deficient (Hygro-3) human breast T47D cell transfectants. GPx activity and GPx1 like antigen concentration in mitochondria were respectively at least 100-fold and 20- to 25-fold higher in GPx2 than Hygro-3 cells. In spite of this large difference in peroxide-scavenging capacity, the basal 8-oxo-dG frequency in mtDNA, assessed by carefully controlled postlabeling assay, was strikingly similar in both cell lines. In contrast, in response to menadione-mediated oxidative stress, induction of 8-oxo-dG and DNA strand breaks was much lower in the GPx1-proficient mitochondria (e.g., +14% 8-oxo-dG versus +54% in Hygro-3 after 1-h exposure to 25 microM menadione, P < 0.05). Our data indicate that the mitochondrial glutathione/GPx1 system protected mtDNA against damage induced by oxidative stress, but did not prevent basal oxidative damage to mtDNA, which, surprisingly, appeared independent of GPx1 status in the T47D model. PMID- 10833430 TI - Angiotensin II modulates ion transport in rat proximal tubules through CYP metabolites. AB - To assess the effect of angiotensin II on ion transport in rat isolated proximal tubules and establish the arachidonic acid cytochrome P450 metabolites' role mediating angiotensin II effect and to analyze whether corticosteroids play a role modulating this effect, we studied the effect of low (10 and 100 pM) and high (0.1-1 microM) angiotensin II concentrations on proximal tubule ion transport, measured as (86)Rb uptake. Low angiotensin II produced a stimulation on the (86)Rb uptake (195.79 +/- 35, 377.9 +/- 81, and 300 +/- 49 pg (86)Rb/microg protein/2 min, for control and 10 and 100 pM angiotensin II, respectively). High angiotensin II concentration inhibited ion transport (0.1 microM, 57.9 +/- 5 and 1 microM, 47.3 +/- 4 pg (86)Rb/microg protein/2 min), this effect was prevented by 17-ODYA and by losartan, while indomethacin had no effect. Dexamethasone treatment increased angiotensin II-induced (86)Rb uptake inhibition and arachidonic acid metabolism (19-, 20-HETE and 12-HETE), while adrenalectomy partly prevented angiotensin II-induced inhibition and decreased cytochrome P450-dependent arachidonic acid metabolism. In conclusion, high doses of angiotensin II produce inhibition of ion transport in rat isolated proximal tubules; this effect is mediated by AT(1) receptors, involves cytochrome P450 dependent arachidonic acid metabolites, and is upregulated by corticosteroids. PMID- 10833431 TI - Induction of the mitochondrial permeability transition in vitro by short-chain carboxylic acids. AB - We recently reported that acrylic acid (AA) induces the MPT in vitro, which we suggested might be a critical event in the acute inflammatory and hyperplastic response of the olfactory epithelium. The purpose of the present investigation was to determine if induction of the MPT is a general response to short-chain carboxylic acids or if there are critical physical chemical parameters for this response. Freshly isolated rat liver mitochondria were incubated in the presence of varying concentrations of selected carboxylic acids. All of the acids that we tested caused a concentration-dependent induction of the MPT, which was blocked by cyclosporine A. Although the C4 carboxylic acids were slightly more potent than the C5 acids, there was no correlation with the degree of saturation, the octanol/water coefficient (log P), or the dissociation constant (pK(a)) of the acids that we tested. We conclude that induction of the MPT in vitro is a general response to short-chain carboxylic acids having a pK(a) of 4 to 5. PMID- 10833432 TI - Identification of novel sequence-specific nuclear factors interacting with mouse senescence marker protein-30 gene promoter. AB - Senescence marker protein-30 (SMP30) gene expression declines during aging in mouse liver. SMP30 also plays a role as Ca(2+)-binding protein localized in cytosol of hepatocytes. To elucidate the molecular mechanism of regulation of SMP30 gene expression we have cloned its gene promoter and carried out DNA protein interaction analyses by DNase I footprinting and electrophoretic mobility shift assay. We have identified a total of eight nuclear factor binding sites within 0.8 kb upstream of transcription start site. Three of these sites are novel DNA sequences with no homology to the existing transcription factor binding site database. Interaction of nuclear factors to these novel cognate sites are DNA sequence-specific. The other five sites correspond to binding sites of known transcription factors, Sp1, AP2, CCAAT box, Lyf-1, and GATA-1. Coordinated orchestration of these factors may contribute to regulation of SMP30 gene expression. PMID- 10833433 TI - 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) induces binding of a 50 kDa protein on the 3' untranslated region of urokinase-type plasminogen activator mRNA. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD), a highly toxic compound that has recently attracted much attention as an environmental contaminant, elicits a variety of toxic responses. Most, if not all, of the toxic effects of TCDD are thought to result from alteration of gene expression. TCDD acts through both transcriptional and posttranscriptional mechanisms to alter gene expression of many genes. Transforming growth factor (TGF)-alpha and urokinase-type plasminogen activator (uPA) are examples of the genes up-regulated posttranscriptionally by TCDD by mRNA stabilization. While effects of TCDD on transcription have been extensively studied, the molecular mechanisms underlying the TCDD-induced changes in mRNA stability are poorly understood. In this study, we investigated the trans acting factors involved in TCDD-dependent mRNA stabilization. UV-crosslinking study showed that a liver cytoplasmic protein of 50 kDa (p50) selectively recognized the 3' UTR of the uPA mRNA in a TCDD-dependent manner. We also showed that the activation of p50 by TCDD is mediated through a protein phosphorylation cascade but not via de novo protein synthesis. This is the first study to show the presence of the TCDD-dependent RNA binding activity which may be involved in TCDD-dependent stabilization of mRNA. PMID- 10833434 TI - Neurocan is a heparin binding proteoglycan. AB - Neurocan and brevican are related chondroitin sulfate proteoglycans which are mainly expressed in the central nervous system. Neurocan and the secreted brevican variant are composed of globular N-terminal hyaluronan binding domains, central O-linked oligosaccharide attachment regions, and globular C-terminal domains. Interaction studies of mouse brain proteoglycans revealed that neurocan, but not brevican, was retained on a heparin affinity matrix. Also a recombinantly produced C-terminal fragment of neurocan, expressed by HEK 293 cells, was retained by the heparin affinity matrix. The substitution of this fragment with a chondroitin sulfate chain did not inhibit binding to the heparin affinity matrix at physiological NaCl concentrations, but decreased the NaCl concentration necessary for elution. Two potential consequences of the heparin binding ability of neurocan are an enforcement of the interaction with other heparin binding molecules and a directed secretion by polarized cells. PMID- 10833435 TI - Mouse and human GTPBP2, newly identified members of the GP-1 family of GTPase. AB - We earlier identified the GTPBP1 gene which encodes a putative GTPase structurally related to peptidyl elongation factors. This finding was the result of a search for genes, the expression of which is induced by interferon-gamma in a macrophage cell line, THP-1. In the current study, we probed the expressed sequence tag database with the deduced amino acid sequence of GTPBP1 to search for partial cDNA clones homologous to GTPBP1. We used one of the partial cDNA clones to screen a mouse brain cDNA library and identified a novel gene, mouse GTPBP2, encoding a protein consisting of 582 amino acids and carrying GTP-binding motifs. The deduced amino acid sequence of mouse GTPBP2 revealed 44.2% similarity to mouse GTPBP1. We also cloned a human homologue of this gene from a cDNA library of the human T cell line, Jurkat. GTPBP2 protein was found highly conserved between human and mouse (over 99% identical), thereby suggesting a fundamental role of this molecule across species. On Northern blot analysis of various mouse tissues, GTPBP2 mRNA was detected in brain, thymus, kidney and skeletal muscle, but was scarce in liver. Level of expression of GTPBP2 mRNA was enhanced by interferon-gamma in THP-1 cells, HeLa cells, and thioglycollate elicited mouse peritoneal macrophages. In addition, we determined the chromosomal localization of GTPBP1 and GTPBP2 genes in human and mouse. The GTPBP1 gene was mapped to mouse chromosome 15, region E3, and human chromosome 22q12-13.1, while the GTPBP2 gene is located in mouse chromosome 17, region C-D, and human chromosome 6p21-12. PMID- 10833436 TI - Interaction of Vesl-1L/Homer 1c with syntaxin 13. AB - Vesl-1S/Homer 1a, reported originally as Vesl/Homer, was isolated as a synaptic plasticity-regulated gene. The expression of Vesl-1S/Homer 1a is regulated during long-term potentiation in the hippocampus. Vesl-1L/Homer 1c, which appears to be formed by a splicing event, shares the N-terminus with Vesl-1S/Homer 1a and also contains additional amino acids at the C-terminus. The short form and the long form of the family members both interact with group 1 metabotropic glutamate receptors (mGluRs). We herein report the identification of syntaxin 13 as a molecule that interacts with Vesl-1L using yeast two-hybrid screening. Syntaxin 13 is a member of the syntaxin family and is regarded as soluble N-ethylmaleimide sensitive attachment proteins receptors (SNAREs) in the endosomal membranes. The interaction of Vesl-1L and syntaxin 13 was biochemically confirmed by in vitro binding assays. The coexpression of the two proteins in the transfected cells resulted in a colocalization in the intracellular vesicle-like structures. We thus propose that the association of Vesl-1L with syntaxin 13 plays a role in the translocation of Vesl-1L to the intracellular organelles. PMID- 10833437 TI - Properties and intracellular localization of calpain activator protein. AB - In this paper, we have further analyzed the properties of calpain activator (CA) in order to better define its physiological function. The activator shows a pH optimum approximately 7.8-8.0, independently of the nature of the buffer used. Although the maximal activity is observed with human acid-denatured globin, the effect of CA is detectable with other protein substrates, such as casein and insulin. A comparable activating effect is observed also with the synthetic substrate Succ-Leu-Tyr-AMC. The activatory effect has been evaluated in a reconstructed system, using plasma membrane Ca(2+)-ATPase as substrate. CA is localized in erythrocyte precursor cells on the inner surface of the plasma membrane in very high amount and its level profoundly decreases up to 10% of the original value when cells reach the terminal differentiated state. PMID- 10833438 TI - From a short amino acidic sequence to the complete gene. AB - A useful strategy directed to the isolation of a required gene with a high GC content is reported. Using a degenerate oligonucleotide probe, deduced from the amino terminus of a protein, it is possible to obtain a fragment of DNA containing its encoding gene by PCR amplification. Furthermore, the cloning of a desired gene can be accomplished in two steps by using an oligonucleotide deduced (i) from an internal sequence, (ii) from a consensus sequence, or (iii) from a DNA sequence adjacent to a disrupting element (transposon, insertion sequence, cassette). This method, which could be applied to a bacteriophage, plasmid, or cosmid genomic library, has been successfully used for cloning several genes from different biological systems. PMID- 10833439 TI - Influence of the histidine tail on the structure and activity of recombinant chlorocatechol 1,2-dioxygenase. AB - We present two efficient expression systems for the chlorocatechol 1, 2 dioxygenase (CCD) from Pseudomonas putida. In the first, CCD (encoded by the clcA gene) was expressed in the pETCLCA vector with the addition of an N-terminal histidine tail. After purification, the enzyme (CCD 6xHis) was proteolytically cleaved with thrombin to remove the His tail. The CD spectra of the cleaved and uncleaved enzymes present only minor differences, indicative of correct protein folding. However, the activity of CCD 6xHis, over a wide range of pH, was typically five times lower. This may be the result of steric hindrance caused by the histidine tail. These data are consistent with results obtained using an alternative construct employing a vector which produces a protein product devoid of the His tail. These results suggest that the His tail may induce subtle effects close to the active site which compromise the recovery of full biological activity. PMID- 10833440 TI - Purification of a novel serine proteinase inhibitor from the skeletal muscle of white croaker (Argyrosomus argentatus). AB - A novel serine proteinase inhibitor has been purified to homogeneity from the skeletal muscle of white croaker (Argyrosomus argentatus). The purification was carried out by ammonium sulfate fractionation, DEAE-Sephacel, heating treatment followed by column chromatographies on SP-Sepharose, Sephadex G-150 and gel filtration high performance liquid chromatography. The molecular mass of the inhibitor was 55 kDa as estimated by SDS-PAGE and gel filtration. It specifically inhibited a myofibril-bound serine proteinase (MBSP) isolated from the skeletal muscle of lizard fish (Saurida wanieso). No inhibition, however, was detected toward other serine proteinases such as bovine trypsin, bovine chymotrypsin and a myofibril-bound serine proteinase from carp (Cyprinus carpio) muscle. Interestingly, the sequences of tryptic digested peptide fragments of MBSPI revealed high identity to that of porcine phosphoglucose isomerase (PGI) (76%) and other PGIs. Furthermore, purified MBSPI exhibits PGI activity, suggesting the inhibitor is a protein closely related to PGI. When rabbit muscle PGI was investigated, it also specifically suppressed the activity of MBSP. It thus strongly suggests that MBSPI is actually PGI and conversely, PGI is a specific inhibitor toward myofibril-bound serine proteinase(s). PMID- 10833441 TI - Distinct regulation of vascular endothelial growth factor in intact human conduit vessels exposed to laminar fluid shear stress and pressure. AB - VEGF is a potent angiogenic factor. We tested the hypothesis that biomechanical forces may regulate VEGF expression. By using a computerized perfusion system, human umbilical veins were exposed to high/low shear stress or intraluminal pressure (25/4 dyn/cm(2) or 40/20 mmHg) for 1.5, 3, or 6 h. Quantification of VEGF gene expression was performed with real-time RT-PCR. VEGF protein was characterized by quantitative immunohistochemistry. All perfusion experiments were performed under identical pH, PO(2), and temperature. Shear stress induced significant biphasic regulation pattern of VEGF (P = 0.0044) with significant downregulation by 45 and 40% after 1.5 and 6 h perfusion, respectively (P = 0.006 and P = 0.0002). The temporal changes of the gene expression were accompanied by synchronal changes at the protein level. High pressure induced transient 25% downregulation of VEGF gene expression after 1.5 h perfusion (P = 0.031). These data provide the first evidence on modulating effects of biomechanical forces on the vascular angiogenic property. PMID- 10833442 TI - Targeting a recombinant adenovirus vector to HCC cells using a bifunctional Fab antibody conjugate. AB - We developed a specific adenoviral gene delivery system with monoclonal antibody (mAb) AF-20 that binds to a 180 kDa antigen highly expressed on human hepatocellular carcinoma (HCC) cells. A bifunctional Fab-antibody conjugate (2Hx 2-AF-20) was generated through AF-20 mAb crosslinkage to an anti-hexon antibody Fab fragment. Uptake of adenoviral particles and gene expression was examined in FOCUS HCC and NIH 3T3 cells by immunofluorescence; beta-galactosidase expression levels were determined following competitive inhibition of adenoviral CAR receptor by excess fibre knob protein. The chimeric complex was rapidly internalized at 37 degrees C, and enhanced levels of reporter gene expression was observed in AF-20 antigen positive HCC cells, but not in AF-20 antigen negative NIH 3T3 control cells. Targeting of recombinant adenoviral vectors to a tumor associated antigen by a bifunctional Fab-antibody conjugate is a promising approach to enhance specificity and efficiency of gene delivery to HCC. PMID- 10833443 TI - ENH, containing PDZ and LIM domains, heart/skeletal muscle-specific protein, associates with cytoskeletal proteins through the PDZ domain. AB - The Enigma homologue protein (ENH), containing an N-terminal PDZ domain and three C-terminal LIM domains, is a heart and skeletal muscle-specific protein that has been shown to preferentially interact with protein kinase C beta (PKCbeta) through the LIM domains (Kuroda et al., J. Biol. Chem. 271, 31029-31032, 1996). We here demonstrate that ENH is colocalized with a cytoskeletal protein alpha actinin in the Z-disk region of rat neonatal cardiomyocytes. Pull-down assays using the glutathione-S-transferase-fusion system also showed the interaction of the PDZ domain of ENH with actin and alpha-actinin. Furthermore, by combined use of the in silico and conventional cDNA cloning methods, we have isolated three ENH-related clones from a mouse heart-derived cDNA library: mENH1 (591 amino acid residues) corresponding to rat ENH, mENH2 (337 residues), and mENH3 (239 residues); the latter two containing only a single PDZ domain. Deciphering their cDNA sequences, these mENH1-3 mRNAs appear to be generated from a single mENH gene by alternative splicing. Northern blot analyses using human cancer cells and mouse embryos have shown expression of each mENH mRNA to vary considerably among the cell types and during the developmental stage. Together with a recent finding that PKCbeta is markedly activated in the cardiac hypertrophic signaling, these results suggest that ENH1 plays an important role in the heart development by scaffolding PKCbeta to the Z-disk region and that ENH2 and ENH3 negatively modulate the scaffolding activity of ENH1. PMID- 10833444 TI - Deregulated expression of homeobox-containing genes, HOXB6, B8, C8, C9, and Cdx 1, in human colon cancer cell lines. AB - Previously we have demonstrated a reciprocal deregulation of various homeobox genes (HOXB6, B8, C8 and C9 vs Cdx-1) in human colorectal cancer (CRC). In the present study, using RT-PCR, we have investigated the expression pattern of these homeobox genes in various human colon cell lines, representing various stages of colon cancer progression and differentiation. Thus, we have tested polyposis coli Pc/AA adenoma cells, Caco-2, HT-29 and LS174T adenocarcinoma cell lines. All cell lines, except LS174T, demonstrated a pattern of deregulated homeobox gene expression which resembled that of CRC. In contrast, the pattern of expression of these genes in the highly oncogenic LS174T cells, as well as in Caco-2 cells transfected with activated Ha-ras or Polyoma middle T oncogene, resembled that of the normal mucosa. The reciprocal deregulation of HOX and Cdx-1 genes in CRC and in CRC-derived cell lines suggests a possible role in human CRC development. PMID- 10833445 TI - Sulfolipid is a potential candidate for annexin binding to the outer surface of chloroplast. AB - Using a subcellular-specific proteomic approach, we have identified by protein microsequencing, a putative 35-kDa annexin from among the chloroplast envelope polypeptides. To confirm this identification, we demonstrate that (a) a 35-kDa protein, identified as annexin by antibody cross-reactivity, co-purifies with Percoll-purified chloroplasts and their envelope membranes when extracted in the presence of Ca(2+) and (b) the native spinach annexin protein binds to chloroplast-specific lipids in a Ca(2+)-dependent manner. The binding of the spinach annexin to these glycerolipids occurs at similar Ca(2+) concentrations as those, which promote the interaction of annexins to phospholipids in other membranes. Among chloroplast glycerolipids known to be accessible on the cytosolic face (outer leaflet) of the outer envelope membrane, sulfolipid, and probably phosphatidylinositol, would be the sole candidates for a putative Ca(2+) dependent interaction of annexin with the chloroplast surface. PMID- 10833446 TI - Selective transcriptional regulations in the human liver cell by hepatitis B viral X protein. AB - The hepatitis B viral X protein (HBx) is known as a transcription factor and potential oncogene. To gain a better view of the effect of HBx on the transcriptional regulation in the human liver cell, we constructed a HepG2 cell line stably expressing HBx (HepG2-HBx), and performed cDNA microarray analysis on 588 cellular cDNAs comparing with untransformed control cells. Two genes (IGFR-2, RhoA) of oncogenes, one gene (p55CDC) of cell cycle regulators, three genes (thrombin receptor, MLK-3, MacMARCKS) of intracellular transducers, one gene (HSP27) of stress response proteins, two genes (FAST kinase, Bak) of apoptosis response proteins, one gene (p21(WAF)) of transcription factors were highly up regulated; one gene (transcription elongation factor SII) of transcription factors and two genes (monocyte chemotactic protein 1, T-lymphocyte-secreted protein I-309) of growth factors were highly down-regulated. These results showed selective transcriptional regulation by HBx in the human liver cell. PMID- 10833447 TI - Determination of genes involved in the process of implantation: application of GeneChip to scan 6500 genes. AB - Using the high-density arrays of oligonucleotides (GeneChip) technology, the expression of uterine genes was examined before and after conceptus implantation in mice. Of the 6500 genes analyzed, levels of 399 gene expressions changed; 192 genes increased levels of expression while the remaining 207 genes declined. The findings suggest that both gene activation and deactivation (suppression) are required for successful implantation. PMID- 10833448 TI - Islet phospholipase A(2) activation is potentiated in insulin resistant mice. AB - Insulin resistance is followed by an islet adaptation resulting in a compensating increase in insulin secretion and hyperinsulinemia. The mechanism underlying this increased insulin secretion is not established. We studied whether islet phospholipase A(2) (PLA(2)) contributes by using C57BL/6J mice fed a high-fat diet, since we previously showed that the insulin responses to the two PLA(2) activating insulin secretagogues carbachol and cholecystokinin (CCK) are enhanced in this model. CCK (100 nM) and carbachol (100 microM) stimulated [(3)H]AA efflux, reflecting PLA(2) activation, both in islets from mice after 12 weeks on high-fat diet and in controls. The efflux increase was more pronounced in islets from high-fat diet-fed mice during both CCK (by 93 +/- 46%; P = 0. 034) and carbachol (by 64 +/- 22%; P = 0.009) stimulation. Also a direct PLA(2) activation by mellitin (2 microg/ml) elicited a potentiated efflux in islets from the insulin-resistant mice (by 361 +/- 107%; P = 0.002). The results suggest that exaggerated non-glucose-induced PLA(2) activation contributes to the islet compensation in insulin resistance. PMID- 10833450 TI - A mechanism for oxygen exchange between ligated oxometalloporphinates and bulk water. AB - Oxygen exchange between high-valent metal-oxo complexes and bulk water has been monitored for nonligated model porphyrins (hemin, FeTDCPPS, MnTMPyP) and the axially ligated microperoxidase-8 (MP-8). Exchange extents up to 90% were measured for MP-8 in spite of the presence of an axial histidine ligand and accompanied by the formation of nonlabelled H(2)O(2) from H(2)(18)O(2). These results point to the existence of a mechanism for oxygen exchange between the high-valent iron-oxo complex and the solvent different from the so-called "oxo hydroxo tautomerism." Regeneration of the primary oxidant, H(2)O(2), and oxygen exchange by axially ligated porphyrins can be explained by a mechanism involving the reversibility of compound I formation. PMID- 10833449 TI - Angiotensin-II subtype 2 receptor agonist (CGP-42112) inhibits catecholamine biosynthesis in cultured porcine adrenal medullary chromaffin cells. AB - Angiotensin II subtype 2 receptor (AT(2)-R) is abundantly expressed in adrenal medullary chromaffin cells. However, the physiological roles of AT(2)-R in chromaffin cells remain to be clarified. Therefore, we investigated the effects of CGP42112 (AT(2)-R agonist) on catecholamine biosynthesis in cultured porcine adrenal medullary cells. We initially confirmed AT(2)-R was predominantly expressed in porcine adrenal medullary cells by [(125)I]-Ang II binding studies. CGP42112 (>==1 nM) significantly inhibited cGMP production from the basal value. Tyrosine hydroxylase (TH) is a rate-limiting enzyme in the biosynthesis of catecholamine, and its activity is regulated by both TH-enzyme activity and TH synthesis. CGP42112 (>==1 nM) significantly inhibited TH-enzyme activity from the basal value. These inhibitory effects of CGP42112 on TH-enzyme activity and-cGMP production were abolished by PD123319 (AT(2)-R antagonist) while CV-11974 (AT(1) R antagonist) was ineffective. We also tested whether decrease of cGMP is involved in the inhibitory effect of CGP42112 on TH-enzyme activity. Pretreatment of 8-Br-cGMP (membrane-permeable cGMP analogue) prevented the inhibitory effect of CGP 42112 on TH-enzyme activity. Similar to that of TH-enzyme activity, CGP42112 (>==1 nM) significantly reduced TH-mRNA and TH-protein level from the basal value, and these inhibitory effects were abolished by PD123319 but not CV 11974. These findings demonstrate that CGP 42112 reduces both TH-enzyme activity and TH-synthesis and that these inhibitory effects could be mediated by decrease of cGMP production. PMID- 10833451 TI - Diverse effect of tributyltin on apoptosis in PC12 cells. AB - It has not been fully elucidated how endocrine-disrupting chemicals disrupt hormone functions or how strong their effects are compared with natural hormones. There is little information concerning the effects of tributyltin (TBT), one of the endocrine disrupters on living organisms. Although TBT at high concentration induced apoptosis in PC12 cells, TBT at low concentration inhibited the DNA fragmentation in the cells cultured in serum-free medium or in medium containing 6-hydroxydopamine. The cell viability grown in both medium conditions increased after treatment with TBT. These findings suggest that TBT exerted a apoptosis inducing and -inhibiting effect. These diverse effect of TBT on apoptosis would cause serious damages on cell differentiation. PMID- 10833452 TI - Full-length cDNA cloning and genomic organization of the mouse liver-specific organic anion transporter-1 (lst-1). AB - We have cloned a cDNA that codes for mouse liver-specific transporter-1, mouse lst-1. The cDNA is comprised of 3296 base pairs and it contains a coding sequence for a protein of 689 amino acids with 12 putative transmembrane domains. The deduced amino acid sequence of the mouse lst-1 shares 64 and 77% identities with the reported human and rat lsts, respectively. Northern blot analysis demonstrates that mouse lst-1 mRNA is expressed exclusively in liver. We also report here the structural organization of the mouse lst-1 gene as the first evidence for the structure of a gene encoding an lst. The mouse lst-1 gene spans approximately 60 kbp in length and consists of 16 exons, including two noncoding exons. All the introns are flanked by GT-AG consensus splice sequences. 5'-Rapid Amplification of cDNA Ends (RACE) analyses demonstrate three splice variant mRNAs involving the noncoding exon 2 and exon 3. The 5'-flanking region of the gene contains consensus CAAT and TATA boxes and several potential binding sites for transcription factors for CAAT enhancer binding protein (C/EBP) and hepatocyte nuclear factors (HNF-3beta, HFH-1, and HFH-2), transcription factors important for liver-specific gene expression. PMID- 10833453 TI - Escherichia coli genome is composed of two distinct types of nucleotide sequences. AB - We calculated correlations of the nucleotide distributions along the E. coli genome. Subsequent cluster analysis of the correlation distributions showed that the genome was composed of two qualitatively different types of nucleotide sequences. The first type exhibited strong correlations of the genomic distributions of A with T and G with C, and high anticorrelations of A with C and G with T. In contrast, the second type was characterized by weak or negligible correlations typical of randomized sequences. Both types of sequences were almost equally abundant in the E. coli genome and their length varied from several hundred nucleotides to about 70 kilobases. They were not disjunct with respect to their (G + C) content but the high correlations and anticorrelations were rather characteristic for (A + T)-rich genomic segments. We offer possible explanations of the mosaic structure of the E. coli genome. PMID- 10833454 TI - An evolutionarily conserved G-protein coupled receptor family, SREB, expressed in the central nervous system. AB - We report here a novel family of G-protein coupled receptor (GPCR) which is extraordinarily conserved among vertebrate species. This family, designated SREB (Super Conserved Receptor Expressed in Brain), consists of at least three members, termed SREB1, SREB2, and SREB3. SREB members share 52-63% amino acid identity with each other and show relatively high similarity to previously known amine amine GPCRs (approximately 25% identity). Amino acid sequence identity between human and rat orthologues is 97% for SREB1 and 99% for SREB3, while the SREB2 sequence is surprisingly completely identical between the species. Furthermore, amino acid sequence of zebrafish SREB2 and SREB3 are 94 and 78% identical to mammal orthologues. Northern blot analysis revealed that SREB members are predominantly expressed in the brain regions and genital organs. Radiation hybrid analysis localized SREB1, SREB2, and SREB3 genes to different human chromosomes, namely 3p21-p14, 7q31 and Xp11, respectively. The high sequence conservation and abundant expression in the central nervous system suggest the existence of undiscovered fundamental neuronal systems consisting of SREB family members and their endogenous ligand(s). PMID- 10833455 TI - First report of the investigation into the importance of pK(a) in the inhibition of estrone sulfatase by sulfamate containing compounds. AB - In an effort to rationalise the inhibitory activity of a range of aminosulfamate compounds and to further investigate the recently reported definitive pharmacophore estrone sulfatase (ES), we undertook extensive synthesis, biochemical evaluation and physicochemical property determination of a range of similar compounds. Here, we report the initial results of our study into a series of simple (aminosulfonate based) substituted phenol derivatives. Using these compounds, we investigated the role of pK(a) in the inhibition of ES. The results of the study suggest that there is a strong correlation between the inhibitory activity and the stability of the resulting O(-) anion (i.e., the pK(a) of the starting phenol). PMID- 10833456 TI - Direct observation of processive movement by individual myosin V molecules. AB - Myosin V is an unconventional myosin thought to move processively along actin filaments. To have hard evidence for the high processivity, we sought to observe directly the movement by individual native chick brain myosin V (BMV) molecules with fluorescent calmodulin. Single BMV molecules did exhibit highly processive movement along actin filaments fixed to a coverslip. BMV continued to move up to the barbed end of its actin track, and did not readily detach from action. The barbed end, therefore, got brighter with time, because of a constant stream of BMV traffic. The maximum speed of the processive movement was 1 microm/s, and the maximum actin-activated ATPase rate was 2.4 s(-1). These values apparently imply that BMV travels a great distance, 400 nm, per an ATPase cycle. PMID- 10833457 TI - Studies on hepatic gene expression in different liver regenerative models. AB - We have investigated the expression of several growth-related genes in the liver after partial hepatectomy in three experimental models: normal, Dexamethasone pretreated, and hypophysectomized rats. Dexamethasone and hypophysectomy resulted in a delay in the peak of cell replication in 6 and 18 h, respectively, when compared to the normal animals. TGFalpha mRNA expression was shifted together with the DNA synthesis, but the expression of c-myc, c-fos, c-jun, HGF, TGFbeta1, IL1beta did not delay. This result suggests that liver-derived TGFalpha but not the other factors are important in the timing of the proliferative response after partial hepatectomy. PMID- 10833458 TI - Unusual substrate specificity of a chimeric hypoxanthine-guanine phosphoribosyltransferase containing segments from the Plasmodium falciparum and human enzymes. AB - Hypoxanthine-guanine phosphoribosyltransferase (HGPRT) catalyzes the phosphoribosylation of hypoxanthine and guanine by transferring the phosphoribosyl moiety from phosphoribosylpyrophosphate (PRPP) on to N9 in the purine base, resulting in the formation of inosine monophosphate (IMP) and guanosine monophosphate (GMP). Xanthine is an additional substrate for the Plasmodium falciparum HGXPRT. Our aim has been to elucidate structural features in HGPRT that govern substrate specificity. We have addressed this problem by engineering chimeric HGPRTs, which contain segments from both the parasite and human enzymes. Four chimeric enzymes were engineered (DS1-DS4), of which the chimeric enzyme, DS1, in which the first 49 residues of human HGPRT were replaced with the corresponding residues from the P. falciparum enzyme, exhibited additional specificity for xanthine. None of the switched residues forms a part of the purine or PRPP binding region in the available crystal structures of HG(X)PRTs. Our data on the chimeric enzyme DS1 provide the first evidence that the N-terminal approximately 50 amino acids, although not proximal to the active site in the crystal structure, can in fact modulate substrate specificity. DS1 exhibits a reduced k(cat) for hypoxanthine and guanine, while its K(m) for these oxopurine bases remains largely unchanged. Its specific activity for xanthine is comparable with hypoxanthine but five times more than that for guanine. PMID- 10833459 TI - Sphingosylphosphorylcholine induces endothelial cell migration and morphogenesis. AB - Sphingosylphosphorylcholine (SPC) is one of the biologically active phospholipids that may act as extracellular messengers. Particularly important is the role of these lipids in the angiogenic response, a complex process involving endothelial cell migration, proliferation, and morphologic differentiation. Here we demonstrate that SPC and its hydrolytic product, sphingosine, induce chemotactic migration of human and bovine endothelial cells. The response is approximately equal to that elicited by vascular endothelial cell growth factor. The effect of SPC and sphingosine was associated with a rapid down-regulation of Edg1, a sphingosine 1-phosphate (SPP)-specific receptor involved in endothelial cell chemotaxis. Both SPC and sphingosine induced differentiation of endothelial cells into capillary-like structures in vitro. Thus, SPC and sphingosine join SPP among the biologically active lipids with angiogenic potential. Since neuronal abnormalities accompany pathological accumulation of SPC in brain tissue, it is possible that SPC is a modulator of angiogenesis in neural tissue upon its release from brain cells following trauma or neoplastic growth. PMID- 10833460 TI - Cloning, characterization, and mapping of the gene encoding the human G protein gamma 2 subunit. AB - G proteins play vital roles in cellular responses to external signals. The specificity of G protein-receptor interaction is mediated mostly by the gamma subunit and the individual members of the gamma-subunit multigene family would hence be expected to each have a particular expression profile. In an experiment designed to isolate genes expressed predominantly in human testis we identified a cDNA fragment corresponding to the gamma2 gene. Although the protein sequence of the gamma2 subunit has previously been published, the cDNA sequence, expression pattern, genomic structure, and localisation of the human GNG2 gene have not been described. We report the complete sequence of the GNG2 cDNA which is 1066 bp long and contains an open reading frame encoding a protein of 71 amino acids. This protein is 100% homologous to the bovine, mouse, and rat G protein gamma2 subunit. The gene structure is very similar to that of other Ggamma-subunit genes in that there are two introns, one located in the 5' UTR and the other within the ORF. We show that this gene is expressed in a range of foetal tissues as well as adult testis, adrenal gland, brain, white blood cells and lung but not in adult liver, muscle, sperm, prostate gland nor in the testes of two different infertile patients. There is evidence that GNG2 is expressed in malignant tissues. Using two independent methods, we have mapped the human GNG2 gene to chromosome 14q21. PMID- 10833461 TI - Isolation and characterization of the human UGT2B7 gene. AB - Glucuronidation is a major pathway involved in the metabolism of drugs and numerous endogenous compounds, such as bile acids and steroid hormones. The enzymes responsible for this conjugation reaction are UDP glucuronosyltransferases (UGT). Among the UGT2B subfamily, UGT2B7, a UGT enzyme present in the liver and several steroid target tissues, is an important member since it conjugates a large variety of compounds including estrogens, androgens, morphine, AZT, and retinoic acid. Although this enzyme is well characterized, the gene encoding the UGT2B7 protein and its promoter region remain unknown. In this article, we report the genomic organization and the promoter region of the human UGT2B7 gene. To isolate this gene, a P-1 artificial chromosome (PAC) library was screened with a full length UGT2B7 probe and a clone of approximately 100 kb in length was isolated. In addition to the UGT2B7 gene, this PAC contains two other UGT2B genes previously characterized, namely UGT2B26P and UGT2B27P. The UGT2B7 gene is composed of six exons spanning approximately 16 kb, with introns ranging from 0.7 to 4.2 kb. The 5'-flanking region of the human UGT2B7 gene contains several potential cis-acting elements such as Oct-1, Pbx-1, and C/EBP. Only one TATA-box at nucleotide -106 was found within the first 500 nucleotides relative to the adenine base of the initiator ATG codon. Characterization of the UGT2B7 gene provides insight into the organization and regulation of this important metabolic gene. PMID- 10833462 TI - Functional expression, purification, and characterization of 3alpha hydroxysteroid dehydrogenase/carbonyl reductase from Comamonas testosteroni. AB - 3alpha-Hydroxysteroid dehydrogenase (3alpha-HSD) catalyzes the oxidoreduction at carbon 3 of steroid hormones and is postulated to initiate the complete mineralization of the steroid nucleus to CO(2) and H(2)O in Comamonas testosteroni. By this activity, 3alpha-HSD provides the basis for C. testosteroni to grow on steroids as sole carbon and energy source. 3alpha-HSD was cloned and overexpressed in E. coli and purified to homogeneity by an affinity chromatography system as His-tagged protein. The recombinant enzyme was found to be functional as oxidoreductase toward a variety of steroid substrates, including androstanedione, 5alpha-dihydrotestosterone, androsterone, cholic acid, and the steroid antibiotic fusidic acid. The enzyme also catalyzes the carbonyl reduction of nonsteroidal aldehydes and ketones such as metyrapone, p-nitrobenzaldehyde and a novel insecticide (NKI 42255), and, based on this pluripotent substrate specificity, was named 3alpha-hydroxysteroid dehydrogenase/carbonyl reductase (3alpha-HSD/CR). It is suggested that 3alpha-HSD/CR contributes to important defense strategies of C. testosteroni against natural and synthetic toxicants. Antibodies were generated in rabbits against the entire 3alpha-HSD/CR protein, and may now be used for evaluating the pattern of steroid induction in C. testosteroni on the protein level. Upon gel permeation chromatography the purified enzyme elutes as a 49.4 kDa protein revealing for the first time the dimeric nature of 3alpha-HSD/CR of C. testosteroni. PMID- 10833463 TI - The renal medullary microcirculation. AB - Blood flow to the renal medulla is supplied through descending vasa recta (DVR), which are derived from the efferent arterioles of juxtamedullary glomeruli. In addition to their role as conduits for blood flow, it is accepted that the vasa recta are countercurrent exchangers. That process, however, involves events which are more complicated than paracellular diffusive exchange of NaCl and urea. Urea transport in DVR is accommodated through the combined expression of endothelial and erythrocyte facilitated carriers while transport of water involves solute driven efflux across water channels. Unlike DVR, which have a continuous endothelium, ascending vasa recta (AVR) are fenestrated with a very high hydraulic conductivity. Transport of water in AVR is probably governed by transmural hydraulic and oncotic pressure gradients. The parallel arrangement of DVR in outer medullary vascular bundles coupled with their capacity for vasomotion implies a role for regulation of the regional distribution of blood flow within the medulla The importance of the latter process in the urinary concentrating mechanism and the exchange of nutrients and O2 is poorly defined. The large number of hormones and autacoids that influence DVR vasomotion, however, suggests that DVR have evolved to optimize the functions of the renal medulla. PMID- 10833464 TI - Caloric restriction and immunosenescence: a current perspective. AB - The age-related decrease in immunologic function is believed to be the major predisposing factor contributing to increased morbidity and mortality with age. Hence, the restoration of immunologic function is expected to have a beneficial effect in reducing pathology and maintaining a healthy condition in advanced age. Among various intervention strategies, caloric restriction (CR) has been shown to be the most powerful modulator of aging process. It is the most efficacious means of increasing longevity and reducing pathology. Several mechanisms have been proposed to explain its beneficial and robust action on various physiological systems, including the immune system. Experimental evidence suggests that CR increases longevity and reduces pathology through its action on the immune system. The observation that CR attenuates immunosenescence has provided a rationale for studying whether CR exerts its action through modulation of gene expression. The available data indicate that the effect of CR on signal transduction and gene expression can vary considerably from gene to gene and from one signaling molecule to another. This review summarizes the studies on the influence of CR on aging immune system and discusses the current state of knowledge on the molecular mechanisms responsible for the immunomodulatory action of caloric restriction. PMID- 10833465 TI - Electrohepatogram in pathologic liver conditions. AB - We have recently studied the electric activity of the liver in both a normal canine model and in humans, and could characterize an electrohepatogram (EHG). Regular and reproducible slow waves or pacesetter potentials (PPs) were recorded. Hepatoarrhythmic electric activity was registered in liver insult of a canine model and in liver cirrhosis. The current communication studied the hepatic electric activity in liver diseases aiming at identifying an EHG for the different pathologic conditions, to be used as an investigative tool in their diagnosis. 55 subjects were studied: 10 with hepatocellular cancer (7 men, 3 women; age 53.7 years), 16 with acute hepatitis (10 men, 6 women; 42.2 years), 14 with metastatic liver deposits (9 men, 5 women; 53.1 years), and 15 healthy volunteers as controls (10 men, 5 women, 43.6 years). Three electrodes were placed 1.5 - 2 cm below and parallel to the costal margin and 4 to 5 cm apart. They were connected to a Beckman R611 recorder. At least two 20-minute recording sessions were performed for each subject. The PPs of the healthy volunteers exhibited a regular rhythm with identical and reproducible frequency and amplitude from the3 electrodes in the same subject. The EHG of the hepatocellular cancer patients showed a 'hepatoarrhythmic' pattern; the waves had irregular frequency and amplitude. In acute hepatitis 2 patterns were encountered: 'silent', which occurred in the preicteric and icteric stages, and 'hepatoarrhythmic' which occurred in the convalescent stage. The liver which had a few metastatic deposits exhibited a hepatoarrhythmic pattern and the liver with numerous deposits presented a 'silent' pattern. The aforementioned patterns were reproducible. In conclusion, different patterns were recorded in the various liver diseases. The patterns seem to reflect the liver cell function. A normal EHG was recorded from the healthy liver, while a silent EHG was registered from the presumably non-functioning liver cells. Hepatoarrhythmia occurred probably when the liver function was impaired. Electrohepatograhy is a simple, easy, non invasive, and non-radiologic procedure. It is suggested to serve as an investigative tool in the diagnosis of liver diseases. PMID- 10833466 TI - Cell cycle implications in the pathogenesis of rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by hyperplasia of the synovial lining cells, angiogenesis, and infiltration of mononuclear cells resulting in pannus formation, cartilage erosion and ultimately joint destruction. Synovial tissue (ST) fibroblast hyperplasia is reminiscent of tumor-like proliferation and is a major cause of cartilage destruction in the RA joint. The RA joint is replete with cytokines and growth factors which exert a synergistic mitogenic effect on ST fibroblasts. As a result, RA ST fibroblasts exhibit elevated gene expression of proto- oncogenes, such as c-Myc, c-Ras, and c Jun and apoptosis inhibitors such as Bcl-2. At the same time, RA ST fibroblasts contain mutations in tumor suppressor genes such as p53. The altered rates of proliferation and apoptosis of RA synovial cells result in the hyperplasia of synovial tissue and in concert with the chronic inflammatory environment ultimately lead to the destruction of the RA joint. PMID- 10833467 TI - Clinical studies of antisense therapy in cancer. AB - The ability to target and inhibit individual gene expression with antisense oligonucleotides has shown promising activity in preclinical cancer models. Recent clinical studies have tested antisense compounds directed against seven cancer related genes including p53, bcl-2, c-raf, H-ras, protein kinase C-alpha, and protein kinase A. Class specific effects of the phosphorothioate backbone common to the first generation of antisense compounds have dominated the side effects of these oligonucleotides. Inhibition of target gene expression has been modest at most, and clinical activity has been primarily anecdotal. Combinations of the antisense compounds with chemotherapy and second-generation oligonucleotides offer promise that these agents might become a standard part of future cancer therapy. PMID- 10833468 TI - Inhaled corticosteroids: a tale of success in childhood asthma. PMID- 10833469 TI - The burden of childhood asthma. AB - Paediatric asthma is a major clinical concern worldwide and represents a huge burden on family and society. It accounts for a large number of lost school days and may deprive the child of both academic achievement and social interaction. Childhood asthma also places strain on healthcare resources as a result of doctor and hospital visits and the cost of treatment. The prevalence of asthma varies worldwide, possibly because of different exposure to respiratory infection, indoor and outdoor pollution, and diet. Certain risk factors appear to predispose children to developing asthma and atopic disease, including incidence and severity of wheezing, atopy, maternal smoking, and number of fever episodes. This paper discusses the burden, prevalence, and risk factors associated with paediatric asthma. PMID- 10833470 TI - The role of inflammation in childhood asthma. AB - The role of inflammation in adult asthma is well known, involving a cascade of immunological stimulation in which mast cells and eosinophils play pivotal roles. However, the assessment of airway inflammation in children is more difficult as the invasive methods used in adults cannot ethically be used for this purpose alone. Nevertheless, limited data from studies using invasive methodology, and studies using novel non-invasive techniques such as sputum induction and nitrous oxide exhalation, are improving knowledge. The immunopathology in childhood asthma appears to mirror that in adult sufferers. The inflammatory processes are evident at an early age in wheezing infants who later develop asthma, and there are different "wheezing phenotypes" in children with atopic asthma or viral associated wheeze. The mechanisms underlying childhood asthma are dependent not only on increased numbers of inflammatory cells in the airways, but also increased activation of these cells. In vitro data have shown that corticosteroids can inhibit the secretion of proinflammatory compounds from alveolar macrophages, suggesting a potential important role for these agents in halting the development of asthma. Techniques for measuring inflammation in infants need to be refined, in order to provide increased knowledge and accurate monitoring of the disease. It is hoped that this will enable the development of early interventions to minimise the impact of asthma in infants who are identified as being susceptible. PMID- 10833471 TI - The role of inhaled corticosteroids in children with asthma. AB - Inhaled corticosteroids offer a wide range of anti-inflammatory activity and have consistently proved to be the most effective medication for the control of childhood asthma. The high efficacy of inhaled corticosteroids has led to their use in milder disease and younger children in the hope that permanent changes in lung function and airway remodelling may be prevented. However, evidence has emerged over the past six years that the first of the inhaled corticosteroids to become available, beclomethasone dipropionate, may cause growth deceleration at a dose of 400 microg per day. This is especially apparent in children with mild symptoms. The newest of the inhaled corticosteroids to be developed, fluticasone propionate, is equipotent to older compounds at half the dose and in low doses is superior in efficacy to sodium cromoglycate. Two recent studies have shown that fluticasone propionate 100-200 microg per day does not cause growth suppression in children with mild asthma. The long term outcome for children who wheeze in early life is difficult to predict. For this reason the use of inhaled corticosteroids in very young children is best reserved for those with severe symptoms or a strong family history of asthma, and evidence, from measurement of inflammatory markers, of airway inflammation. PMID- 10833472 TI - The GI core curriculum: five years later. PMID- 10833473 TI - Claudins regulate the intestinal barrier in response to immune mediators. AB - BACKGROUND & AIMS: To determine the functional role of immune mediators in the formation of the intestinal barrier, we have examined the regulation of claudin expression by interleukin (IL)-17 in human intestinal epithelial cells. METHODS: Expression of claudins, extracellular signal-related (ERK) mitogen-activated protein kinases (MAPKs), and activated ERK MAPKs was determined by immunoblotting. Claudin membrane association was assessed by immunohistochemistry and claudin messenger RNA expression by Northern blot analysis. Intestinal epithelial barrier function was characterized through transepithelial electrical resistance and mannitol tracer flux. RESULTS: IL-17 induced the development of a paracellular barrier of T84 cell monolayers. Inhibition of ERK activation with the MEK inhibitor PD98059 blocked IL-17 as well as basal development of tight junctions in T84 cells. IL-17 induced formation of tight junctions correlated with up-regulation of claudin-1 and claudin-2 gene transcription. Inhibition of MEK reduced the activated and basal expression of claudin-2 messenger RNA and protein expression. Functional MEK was required for the expression and membrane association of claudin-2 but not claudin-1 in T84 cells. CONCLUSIONS: MEK activity is required for claudin-mediated formation of tight junctions. IL-17 is able to regulate the intestinal barrier through the ERK MAPK pathway. PMID- 10833474 TI - Par-4, a proapoptotic gene, is regulated by NSAIDs in human colon carcinoma cells. AB - BACKGROUND & AIMS: Many reports indicate that nonsteroidal anti-inflammatory drugs (NSAIDs) have antineoplastic effects, but the precise molecular mechanism(s) responsible are unclear. We evaluated the effect of cyclooxygenase (COX) inhibitors (NSAIDs) on human colon carcinoma cells (HCA-7) and identified several genes that are regulated after treatment with NS-398, a selective COX-2 inhibitor. METHODS: Differential display polymerase chain reaction cloning techniques were used to identify genes regulated by treatment with NSAIDs and selective COX-2 inhibitors. RESULTS: A prostate apoptosis response 4 (Par-4) gene was up-regulated after NSAID treatment. Par-4 was first isolated from prostate carcinoma cells undergoing apoptosis, and expression of Par-4 sensitized cancer cells to apoptotic stimuli. Par-4 levels were increased in cells treated with COX inhibitors such as NS-398, nimesulide, SC-58125, and sulindac sulfide. Treatment of HCA-7 cells with these agents also induced apoptotic cell death. CONCLUSIONS: The results suggest that regulation of Par-4 contributes to the proapoptotic effects of high-dose COX inhibitors (NSAIDs) by serving as a downstream mediator leading to initiation of programmed cell death. PMID- 10833475 TI - Azathioprine for maintenance of remission in Crohn's disease: benefits outweigh the risk of lymphoma. AB - BACKGROUND & AIMS: Azathioprine is a commonly used and effective treatment for maintenance of remission for patients with steroid-dependent Crohn's disease (CD). However, azathioprine therapy is associated with an increased risk of non Hodgkin's lymphoma. The objective of this analysis was to determine the impact of azathioprine therapy on survival and quality-adjusted life expectancy after accounting for both the benefits of therapy and potential increased risk of lymphoma. METHODS: A decision analysis using a Markov model depicting the natural history of alternative management strategies for maintenance of remission in patients with CD was performed. RESULTS: In the base-case analysis, treatment of CD patients with a steroid-induced remission with azathioprine resulted in an average increase in life expectancy of 0.04 years and 0.05 quality-adjusted years. The incremental gain in life expectancy decreased with increasing patient age and increasing risk of lymphoma. CONCLUSIONS: Therapy with azathioprine to preserve remission in patients with CD results in increased quality-adjusted life expectancy. This increase was greatest in young patients who have the lowest baseline risk of non-Hodgkin's lymphoma and who have the greatest life expectancy in the absence of a CD-related death. PMID- 10833476 TI - Genotypic analysis of thiopurine S-methyltransferase in patients with Crohn's disease and severe myelosuppression during azathioprine therapy. AB - BACKGROUND & AIMS: Myelosuppression in patients with Crohn's disease (CD) treated with azathioprine has been attributed to low activity of thiopurine S methyltransferase (TPMT). Allelic variants of the TPMT gene responsible for changes in the enzyme activity have been characterized. We investigated the distribution of mutant alleles associated with TPMT deficiency in patients with CD and myelosuppression during azathioprine/6-mercaptopurine therapy. METHODS: Forty-one patients with CD were included. They developed leukopenia or thrombocytopenia during azathioprine or 6-mercaptopurine treatment. Polymerase chain reaction-based methods were used to search for mutations associated with TPMT deficiency. RESULTS: Four patients (10%) had 2 mutant alleles associated with TPMT deficiency, 7 (17%) had 1 mutant allele, and 30 (73%) had no known TPMT mutation. The delay between administration of the drug and occurrence of bone marrow toxicity was less than 1.5 months in the 4 patients with 2 mutant alleles, and ranged from 1 to 18 months in patients with 1 mutant allele and from 0.5 to 87 months in patients with normal genotype. CONCLUSIONS: Twenty-seven percent of patients with CD and myelosuppression during azathioprine therapy had mutant alleles of the TPMT gene associated with enzyme deficiency. Myelosuppression is more often caused by other factors. Continued monitoring of blood cell counts remains mandatory in patients treated with azathioprine. PMID- 10833477 TI - Epstein-Barr virus-associated gastric carcinomas: relation to H. pylori infection and genetic alterations. AB - BACKGROUND & AIMS: The association of Epstein-Barr virus (EBV) and gastric carcinomas (GCs) has been shown to vary among different populations and certain histological subtypes. Few studies have addressed the status of Helicobacter pylori infection and genetic alterations in these EBV-positive or -negative GCs. METHODS: Eleven gastric lymphoepithelioma-like carcinomas (LELCs) and 139 cases of common non-LELCs were evaluated for the presence of EBV DNA using polymerase chain reaction (PCR) and RNA in situ hybridization. H. pylori infection was determined by anti-H. pylori immunoglobulin G in preoperative sera. Immunostaining for p53, c-erbB-2, and E-cadherin was performed. Microsatellite instability was analyzed by PCR using 10 primers. RESULTS: EBV was detected in 11 (100%) LELCs and in 19 (13.7%) of 139 common GCs. Compared with EBV-negative GCs, gastric LELCs tended to have a relatively higher frequency of proximal location, diffuse histological subtype, p53 overexpression, and reduced E-cadherin expression but a lower frequency of lymph node metastasis, previous H. pylori infection, and c-erbB-2 overexpression. In contrast, no significant difference of clinicopathologic and genetic profiles was observed between EBV-positive non-LELC GCs and EBV-negative GCs. No correlation of microsatellite instability was found among these 3 subsets of GCs. CONCLUSIONS: Dissecting clinicopathologic characteristics and infection status of EBV and H. pylori provide additional evidence of etiological and genetic heterogeneity for GC. Distinct clinicopathologic and genetic pathways exist in gastric LELCs, in which EBV may play a more important role than H. pylori infection. PMID- 10833478 TI - Alterations in exon 4 of the p53 gene in gastric carcinoma. AB - BACKGROUND & AIMS: Our long-term goal was to evaluate the role of p53 in the prognosis of gastric cancer. We previously showed a discrepancy between p53 expression and the presence of mutations when only exons 5-9 were examined. We then evaluated exon 4. METHODS: DNA was sequenced from 217 gastric cancers to detect exon 4 alterations. Codon 72 was examined by restriction enzyme digestion. RESULTS: Mutations were present in 3.2% of tumors. In addition, 2 polymorphic sites were found at codons 36 and 72. Polymorphisms at codon 36 were only found in 2 patients. In contrast, the codon 72 polymorphism was very frequent. The genotype frequency was arg/arg (54%), arg/pro (33%), and pro/pro (14%). The genotype of the polymorphic site varied with race (P = 0.001): 64% of whites had the arg/arg genotype, compared with 24% of blacks. The difference in genotype by site, sex, or histological tumor type was not statistically significant (P = 0.067). CONCLUSIONS: There are several exon 4 alterations in gastric cancers. These include the rare mutations and the very rare codon 36 polymorphism. The most common change is the codon 72 polymorphism, the genotype of which differs significantly with race. The more common arg/arg genotype in whites may explain why whites are more prone to develop cardiac cancer, whereas the more common proline allele in blacks may explain why they are more prone to develop antral cancers. Further studies are required to determine whether the codon 72 polymorphism affects patient predisposition to gastric cancer. PMID- 10833479 TI - Activation of c-K-ras mutations in human gastrointestinal tumors. AB - BACKGROUND & AIMS: Ras genes are the most frequently detected oncogenes in human malignancies. Data regarding the frequency of c-K-ras mutations in esophageal, gastric, and small bowel tumors are limited and controversial. METHODS: DNA was extracted from 262 formalin-fixed, paraffin-embedded sections of gastrointestinal samples and tumors, including Barrett's esophagus, esophageal squamous cell carcinomas and adenocarcinomas, and small and large bowel adenomas and adenocarcinomas. The presence of c-K-ras codon 12 mutations was determined using a nonradioactive polymerase chain reaction-based restriction fragment length polymorphism assay. RESULTS: c-K-ras mutations were detected in 1 of 39 (2%) patients with Barrett's esophagus, 1 of 21 (5%) adenocarcinomas, 0 of 27 squamous cell carcinomas of the esophagus, and 1 of 32 (3%) gastric adenocarcinomas. It was also present in 8 of 20 (40%) and 10 of 28 (36%) small bowel adenomas and adenocarcinomas, respectively. Similar numbers were observed in 10 of 25 (40%) large bowel adenomas and 11 of 30 adenocarcinomas (37%). Mutations were not associated with age, gender, histology, grade, stage, location, or mortality. CONCLUSIONS: The frequency of codon 12 c-K-ras mutations in small and large bowel tumors is approximately 10-fold higher than that of tumors in the upper gastrointestinal tract. PMID- 10833480 TI - Identification of transport abnormalities in duodenal mucosa and duodenal enterocytes from patients with cystic fibrosis. AB - BACKGROUND & AIMS: The duodenum is a cystic fibrosis transmembrane conductance regulator (CFTR)-expressing epithelium with high bicarbonate secretory capacity. We aimed to define the role of CFTR in human duodenal epithelial bicarbonate secretion in normal (NL) subjects and patients with cystic fibrosis (CF). METHODS: Endoscopic biopsy specimens of the duodenal bulb were obtained from 9 CF patients and 16 volunteers. Tissues were mounted in modified Ussing chambers. Bicarbonate secretion and short-circuit current (Isc) were quantitated under basal conditions and in response to dibutyryl adenosine 3',5'-cyclic monophosphate (db-cAMP), carbachol, and the heat-stable toxin of Escherichia coli (STa). Duodenocytes were also isolated and loaded with the pH-sensitive fluoroprobe BCECF/AM, and intracellular pH (pH(i)) was measured at rest and after intracellular acidification and alkalinization. RESULTS: Basal HCO(3)(-) secretion and Isc were significantly lower in the CF vs. NL duodenal mucosa. In contrast to NL, db-cAMP failed to alter either HCO(3)(-) or Isc in CF tissues. However, in CF, carbachol resulted in an electroneutral HCO(3)(-) secretion, whereas STa induced electrogenic HCO(3)(-) secretion that was similar to NL. In CF and NL duodenocytes, basal pH(i) and recovery from an acid load were comparable, but pH(i) recovery after an alkaline load in CF duodenocytes was Cl( ) dependent, whereas in NL duodenocytes it was Cl(-) independent. CONCLUSIONS: These findings implicate CFTR in NL duodenal alkaline transport and its absence in CF. Although duodenal bicarbonate secretion is impaired in CF tissues, alternate pathway(s) likely exist that can be activated by carbachol and STa. PMID- 10833481 TI - Constitutive and regulated secretion of secretory leukocyte proteinase inhibitor by human intestinal epithelial cells. AB - BACKGROUND & AIMS: Epithelial cells participate in immune regulation and mucosal integrity by generating a range of biologically active mediators. In the intestine, little is known about the potential endogenous anti-inflammatory molecules. Secretory leukocyte proteinase inhibitor (SLPI) is a major serine proteinase inhibitor, a potent antibiotic, and thus a potential anti-inflammatory molecule, although it is not known if it is secreted by intestinal epithelial cells. METHODS: We show, by reverse-transcription polymerase chain reaction, the presence of SLPI messenger RNA in human model intestinal epithelial cell lines (Caco2-BBE, T84, and HT29-Cl.19A) and human jejunum and colon biopsy specimens. The polymerase chain reaction product was cloned and sequenced and is identical to that of SLPI isolated previously from the human parotid gland. RESULTS: As analyzed by enzyme-linked immunosorbent assay, the constitutive secretion of SLPI occurs in a markedly polarized manner toward the apical surface and is enhanced by inflammatory mediators including tumor necrosis factor alpha and interleukin 1beta (approximately 3.5-fold increase over control value). SLPI release is also stimulated by activation of protein kinase C isoenzymes, but not by activation of adenosine 3',5'-cyclic monophosphate- or Ca(2+)-regulated signaling molecules. SLPI protein is detectable in intestinal lavage fluids collected from normal adult humans. Recombinant SLPI attenuates digestive enzyme (trypsin)- or leukocyte proteinase (elastase)-induced permeability alteration of a model epithelia in a dose-dependent manner. Moreover, SLPI exhibits an antibacterial activity against at least one major intestinal pathogen, Salmonella typhimurium. In contrast, SLPI does not influence epithelial barrier integrity as assessed by transepithelial conductance measurements or electrogenic ion transport. CONCLUSIONS: These results establish that human intestinal epithelium expresses and apically secretes SLPI, a molecule that may significantly contribute to the protection against attack from inflammatory cells and digestive enzymes, as well as against microbial infection. PMID- 10833482 TI - Inhibition of gastric mucin synthesis by Helicobacter pylori. AB - BACKGROUND & AIMS: Mucins are high-molecular-weight glycoproteins that protect the gastric epithelium. Previous data suggested that gastric surface-type mucin is decreased in Helicobacter pylori-infected patients and restored after eradication of the infection. Our aim was to determine the effect of H. pylori on mucin synthesis in cultured gastric epithelial cells. METHODS: Mucin synthesis was measured by labeling with [(3)H]glucosamine and size-exclusion chromatography. Expression of MUC5AC and MUC1 mucin protein antigens was quantitated by Western blot analysis. RESULTS: Mucin synthesis was inhibited more than 80% when KATO III cells were incubated with H. pylori, with no effect on mucin secretion or degradation. Inhibition was rapid (4 hours), partially reversible, dependent on concentration of bacteria, and associated with the insoluble membrane fraction. H. pylori decreased levels of MUC5AC and MUC1 mucins. MUC1 inhibition was half-maximal by 4 hours and partially reversed by 24 hours, but the decrease in MUC5AC was less rapid and not reversible within 24 hours. CONCLUSIONS: H. pylori inhibits total mucin synthesis in vitro and decreases the expression of MUC5AC and MUC1. A decrease in gastric mucin synthesis in vivo may disrupt the protective surface mucin layer. PMID- 10833483 TI - Reversible drug-induced oxyntic atrophy in rats. AB - BACKGROUND & AIMS: Oxyntic atrophy is the hallmark of chronic gastritis. Many studies have sought to develop animal models for oxyntic atrophy, but none of them are reversible. We now report that rats administered high doses of DMP 777 demonstrate reversible oxyntic atrophy. METHODS: DMP 777 was administered to CD-1 rats by oral gavage (200 mg. kg(-1). day(-1)). Serum gastrin level, in vivo acid secretion, and gastric histological changes were evaluated in DMP 777-dosed animals. Direct effects of DMP 777 on parietal cells were evaluated by assessment of aminopyrine accumulation into isolated rabbit parietal cells, as well as by assessment of DMP 777 effects on acridine orange fluorescence and H(+),K(+) adenosine triphosphatase (ATPase) activity in isolated tubulovesicles. RESULTS: Oral dosing with DMP 777 caused a rapid increase in serum gastrin levels and severe hypochlorhydria. DMP 777 inhibited aminopyrine accumulation into rabbit parietal cells stimulated with either histamine or forskolin. DMP 777 reversed a stimulated proton gradient in isolated parietal cell tubulovesicles. Oral dosing with DMP 777 led to rapid loss of parietal cells from the gastric mucosa. In response to the acute loss of parietal cells, there was an increase in the activity of the progenitor zone along with rapid expansion of the foveolar cell compartment. DMP 777 treatment also led to the emergence of bromodeoxyuridine labeled cells and cells positive for periodic acid-Schiff in the basal region of fundic glands. With extended dosing over 3-6 months, foveolar hyperplasia and oxyntic atrophy were sustained while chief cell, enterochromaffin-like cell, and somatostatin cell populations were decreased. No histological evidence of neoplastic transformation was observed with dosing up to 6 months. Withdrawal of the drug after 3 or 6 months of dosing led to complete restitution of the normal mucosal lineages within 3 months. CONCLUSIONS: DMP 777 acts as a protonophore with specificity for parietal cell acid-secretory membranes. DMP 777 in high doses leads to the specific loss of parietal cells. Foveolar hyperplasia, loss of normal gland lineages, and the emergence of basal mucous cells appear as sequelae of the absence of parietal cells. The results suggest that parietal cells are critical for the maintenance of the normal mucosal lineage repertoire. PMID- 10833484 TI - Antibiotic therapy attenuates colitis in interleukin 10 gene-deficient mice. AB - BACKGROUND & AIMS: Interleukin (IL)-10 gene-deficient mice, raised under germfree conditions, do not develop colitis, implying a role for bacteria. This study mapped the appearance of luminal colonic bacteria and, using antibiotic treatment, determined their association with colitis in IL-10 gene-deficient mice. METHODS: Mice were treated with ciprofloxacin or with neomycin and metronidazole. The intestine was harvested for histological scoring and bacterial assessment. RESULTS: At 2 weeks of age, before the development of colitis, IL-10 gene-deficient mice demonstrated an earlier appearance of Streptococcus and Clostridium sp., and had a greater proportion (P < 0.01) of bacteria adherent to the colonic mucosa. This pattern of increased adherent bacteria persisted for the 12 weeks of study. Treatment of mice before the onset of colonic inflammation, with either antibiotic regime, reduced mucosal adherent bacteria and prevented colitis (P < 0.01). In contrast, treatment of established colitis with neomycin and metronidazole did not reduce adherent bacterial levels, yet was more efficacious (P < 0.05) in treating established colitis than ciprofloxacin, which did reduce adherent colonic bacteria. CONCLUSIONS: In the IL-10 gene-deficient mouse model, the appearance and number of mucosal adherent colonic bacteria are altered before the onset of colitis. Antibiotics both prevent and treat the colitis through correction of this primary bacterial alteration. PMID- 10833485 TI - Lipopolysaccharide is radioprotective in the mouse intestine through a prostaglandin-mediated mechanism. AB - BACKGROUND & AIMS: The bone marrow and the intestine are the major sites of radiation-induced injury. The cellular response to radiation injury in the intestine or bone marrow can be modulated by agents given before irradiation. Lipopolysaccharide is known to be radioprotective in the bone marrow, but its effect on the intestine is not known. We sought to determine if lipopolysaccharide is radioprotective in the intestine and, if so, to determine the mechanism of its radioprotective effects. METHODS: Mice were treated with parenteral lipopolysaccharide or vehicle and then irradiated (14 Gy total body irradiation in a cesium irradiator). The number of surviving intestinal crypts was assessed 3.5 days after irradiation using a clonogenic assay. RESULTS: Parenteral administration of lipopolysaccharide 2-24 hours before irradiation resulted in a 2-fold increase in the number of surviving crypts 3.5 days after irradiation. The radioprotective effects of lipopolysaccharide could be eliminated by coadministration of a selective inhibitor of cyclooxygenase 2. Lipopolysaccharide was radioprotective in wild-type mice but not in mice with a disrupted cyclooxygenase 2. Parenteral administration of lipopolysaccharide resulted in increased production of prostaglandins in the intestine and in the induction of cyclooxygenase 2 expression in subepithelial fibroblasts and in villous, but not crypt, epithelial cells. CONCLUSIONS: Lipopolysaccharide is radioprotective in the mouse intestine through a prostaglandin-dependent pathway. PMID- 10833486 TI - Liver fibrosis in overweight patients. AB - BACKGROUND & AIMS: A common clinical issue is whether overweight patients with abnormal liver function test results should undergo liver biopsy. Although serious liver injury can occur, its prevalence and risk factors are not well known. METHODS: Ninety-three consecutive patients with abnormal liver function tests (but without overt liver disease), body mass index (BMI) > 25 kg/m(2), and no alcoholic, viral, autoimmune, drug-induced, or genetic liver disease were retrospectively studied. Clinical, biological, and histological variables were tested for association with septal fibrosis or cirrhosis. RESULTS: Septal fibrosis was present in 28 patients (30%) including cirrhosis in 10 (11%). Age >/= 50 years (odds ratio [OR], 14.1), BMI >/= 28 kg/m(2) (OR, 5.7), triglycerides >/= 1.7 mmol/L (OR, 5), and alanine aminotransferase (ALT) >/= 2N (OR, 4.6) were independently associated with septal fibrosis. Among histological features, septal fibrosis was strongly associated with necroinflammatory activity (OR, 44). A score combining age, BMI, triglycerides, and ALT had 100% negative predictive value for septal fibrosis when scoring 0 or 1 (100% sensitivity for a specificity of 47%). CONCLUSIONS: Septal fibrosis occurs frequently in overweight patients with abnormal liver function tests. A clinicobiological score combining BMI, age, ALT, and triglycerides could improve selection of patients for liver biopsy. PMID- 10833487 TI - The MHC is a major determinant of viral status, but not fibrotic stage, in individuals infected with hepatitis C. AB - BACKGROUND & AIMS: In hepatitis C infection, several studies have examined the role of the major histocompatibility complex (MHC) in determining outcome, with variable results. To clarify the importance of MHC, we examined class II DR and DQ antigens in a homogenous cohort of women exposed to hepatitis C genotype 1b from a single inoculum. METHODS: Of 243 participants, 95 had spontaneous viral clearance and 148 are chronically infected. The frequencies of HLA class II DR and DQ antigens were compared between the 2 groups and between liver biopsy findings of 145 chronically infected subjects. RESULTS: DRB1*0101 and DQB1*0501 alleles were more frequent in subjects who sustained viral clearance than in chronically infected subjects (32.3% and 36.8% vs. 8.8% and 14.2%, respectively; P = 0.002). DRB1*03011 and DQB1*0201 occurred more frequently in chronically infected subjects than in those who cleared the virus (41.5% and 42.6% vs. 16.7% and 15.8%, respectively; P = 0.001). Both DRB1*03011 and DQB1*0201 were significantly less frequent in those with higher inflammatory scores on liver biopsy. CONCLUSIONS: We show that in a homogenous cohort of women infected with the same hepatitis C virus, several HLA antigens are associated with either viral clearance or persistence. This suggests a strong role for host immunogenetic factors in determining outcome in hepatitis C infection. PMID- 10833488 TI - Splanchnic and leg exchange of amino acids and ammonia in acute liver failure. AB - BACKGROUND & AIMS: In patients with acute liver failure, hyperammonemia is associated with cerebral herniation. We examined the splanchnic and leg exchange of amino acids, urea, and ammonia in such patients. METHODS: Bedside liver vein catheterization was used in 22 patients after development of hepatic encephalopathy grades III-IV. Femoral venous blood was sampled in 7 of these patients. RESULTS: Arterial amino acid concentration (8.1 +/- 4.1 mmol/L) was increased 4-fold above normal. Glutamine (2.4 +/- 1.8 mmol/L) and alanine (0.57 +/- 0.35 mmol/L) were by far the predominant amino acids exchanged in the splanchnic and leg circulation. In the splanchnic circulation, there was a net uptake of glutamine (241 +/- 353 micromol/min) and ammonia and alanine were released in an almost 1:1 stoichiometry (r(2) = 0.47; P < 0.001). In the leg, ammonia and alanine were removed and glutamine released. The leg ammonia concentration difference was correlated to that of glutamine (r(2) = 0.80; P = 0.008) and alanine (r(2) = 0.67; P = 0.03). CONCLUSIONS: Splanchnic metabolism of glutamine in combination with decreased hepatic function was responsible for the splanchnic release of ammonia and alanine. These processes were reversed in skeletal muscle. Stimulation of skeletal muscle metabolism of ammonia could be a important target for future treatment of patients with acute liver failure. PMID- 10833489 TI - Enhanced urinary excretion of cysteinyl leukotrienes in patients with acute alcohol intoxication. AB - BACKGROUND & AIMS: Leukotrienes are proinflammatory mediators. Ethanol inhibits the catabolism of both cysteinyl leukotrienes (leukotriene E(4) [LTE(4)] and N acetyl-LTE(4)) and leukotriene B(4) (LTB(4)) in hepatocytes. We examined the metabolic derangement of leukotriene inactivation by ethanol in humans in vivo. METHODS: LTE(4), N-acetyl-LTE(4), LTB(4), and 20-hydroxy-LTB(4) were quantified in urine samples from 16 patients with acute alcohol intoxication (mean blood ethanol, 75 mmol/L). In 9 healthy volunteers, urinary LTE(4) was determined before and after ethanol consumption (mean blood ethanol, 14 mmol/L). RESULTS: The excretion of LTE(4) during alcohol intoxication was 286 compared with 36 nmol/mol creatinine in healthy subjects (P < 0.01); the corresponding values for N-acetyl-LTE(4) were 101 and 11 nmol/mol creatinine, respectively (P < 0.001). This excretion of cysteinyl leukotrienes decreased when the blood ethanol concentration returned to normal. LTB(4) and 20-hydroxy-LTB(4) were detectable only in patients with excessive blood ethanol concentrations (mean, 95 mmol/L). In healthy volunteers, LTE(4) excretion increased 3-5 hours after ethanol consumption (mean peak concentration of 1.5 nmol/L compared with 0.5 nmol/L for basal values; P < 0.005). CONCLUSIONS: Ethanol at high concentration induces increased leukotriene excretion into urine. These changes are consistent with inhibition of leukotriene catabolism and inactivation induced by ethanol, as well as with a higher leukotriene formation caused by ethanol-induced endotoxemia. PMID- 10833490 TI - Angiotensin II induces contraction and proliferation of human hepatic stellate cells. AB - BACKGROUND & AIMS: Circulating levels of angiotensin II (ANGII), a powerful vasoconstrictor factor, are frequently increased in chronic liver diseases. In these conditions, hepatic stellate cells (HSCs) proliferate and acquire contractile properties. This study investigated the presence of receptors for ANGII and the effects of ANGII in human HSCs activated in culture. METHODS: The presence of ANGII receptors was assessed by binding studies. The effects of ANGII on intracellular calcium concentration ([Ca(2+)](i)), cell contraction, and cell proliferation were also assessed. RESULTS: Binding studies showed the presence of ANGII receptors of the AT1 subtype. ANGII elicited a marked dose-dependent increase in [Ca(2+)](i) and cell contraction. Moreover, ANGII stimulated DNA synthesis and increased cell number. All these effects were totally blocked by losartan and reduced by nitric oxide donors or prostaglandin E(2). The effects of ANGII were barely detectable in quiescent cells (2 days in culture), suggesting that phenotypic transformation of HSCs is associated with a marked increase in the effects of ANGII. CONCLUSIONS: ANGII induces contraction and is mitogenic for human-activated HSCs by acting through AT1 receptors. These results suggest that activated HSCs are targets of the vasoconstrictor action of ANGII in the intrahepatic circulation. PMID- 10833491 TI - Bile duct ligation in rats induces biliary expression of cytokine-induced neutrophil chemoattractant. AB - BACKGROUND & AIMS: Bile duct obstruction causes neutrophilic inflammation of the liver and leads to hepatic fibrosis. In obstructive liver disease, the localization of neutrophils in portal tracts suggests that cells within this region produce neutrophil chemoattractants. In this study, we investigated whether bile duct obstruction in rats induces portal expression of cytokine induced neutrophil chemoattractant (CINC). METHODS: Rats underwent bile duct ligation for 3 hours to 8 days. CINC regulation was examined in vivo at various intervals by immunohistochemistry, ribonuclease protection, and in situ hybridization. CINC production was also investigated in cell culture, in response to putative stimuli from obstructed liver. RESULTS: Bile duct ligation caused neutrophilic infiltration of the liver within 3 hours. CINC was also rapidly induced, with specific expression identified in biliary cells. Rat intrahepatic biliary cells produced CINC constitutively in culture; when exposed to cholestatic bile, they showed a 12-fold increase in CINC secretion. The effect of bile was not attributable to toxicity or to dissolved cytokines or endotoxin. Mechanical strain, designed to mimic the stretching of biliary cells during obstruction, did not induce CINC. CONCLUSIONS: Biliary cells contribute to hepatic inflammation during cholestasis by producing neutrophil chemoattractants. A major stimulus to biliary chemoattractant production in vivo may be bile itself. PMID- 10833492 TI - An oral endothelin-A receptor antagonist blocks collagen synthesis and deposition in advanced rat liver fibrosis. AB - BACKGROUND & AIMS: Endothelin 1 induces contraction, proliferation, and collagen synthesis of hepatic stellate cells in vitro, which may be mediated via the endothelin A receptor. It is unknown if specific blockade of the endothelin A receptor inhibits hepatic fibrosis in vivo. METHODS: Groups of 10-20 rats with bile duct occlusion were treated with the nonpeptide endothelin-A receptor antagonist LU 135252 at 80 mg. kg(-1). day(-1) from week 1-6 or from week 4-6, or with LU at 10 mg. kg(-1). day(-1) from week 1-6. Animals with bile duct occlusion alone and sham-operated rats without or with LU at 80 mg. kg(-1). day(-1) over 6 weeks served as controls. After 6 weeks, parameters of fibrogenesis were determined. RESULTS: LU treatment led to improved histology, paralleled by a dose dependence up to 60% reduction of liver collagen, even when administered at an advanced fibrosis stage. This was accompanied by a decreased messenger RNA of hepatic procollagen alpha1(I) and tissue inhibitor of metalloproteinase 1, 2 major effectors of fibrosis, and of serum procollagen type III, a surrogate marker of liver fibrogenesis. CONCLUSIONS: Selective endothelin-A receptor blockade can dramatically reduce collagen accumulation in rat secondary biliary fibrosis, a model refractory to most potential antifibrotic agents. Endothelin-A receptor antagonists are promising antifibrotic agents in chronic liver disease. PMID- 10833493 TI - Role of biliary phosphatidylcholine in bile acid protection and NSAID injury of the ileal mucosa in rats. AB - BACKGROUND & AIMS: We explored the role of biliary phosphatidylcholine (PC) in protection of the intestinal mucosa against bile salt (BS)-induced intestinal injury and how this property may be blocked by indomethacin (Indo), a nonsteroidal anti-inflammatory drug (NSAID) that is secreted into the bile. METHODS: We performed in vivo studies in which bile was collected over a 2-hour period after rats were intragastrically administered Indo (25 mg/kg) or an equivalent volume of saline (controls). The bile samples (some of which were supplemented with PC) were then instilled into a loop of distal ileum of anesthetized rats. After a 30-minute exposure period, we measured the hemoglobin concentration of the ileal loop fluid, as an index of bleeding, and mucosal contact angles, as an index of surface hydrophobicity. A similar in vivo experiment was performed in which model bile containing 5 mmol/L each of the BS, sodium deoxycholate, PC, or Indo, alone and in combination, was instilled into ileal loops. In our in vitro test system, human erythrocytes were exposed to the above biliary constituents, and hemolysis was measured spectrophotometrically. RESULTS: Bile from Indo-pretreated rats decreased the surface hydrophobicity and induced bleeding of ileal loops in comparison with control bile, and both NSAID induced changes were reversed if PC was added to the bile. Similarly, synthetic BS caused gastrointestinal bleeding, decreased ileal contact angles, and induced erythrocyte hemolysis, all of which were reversed by addition of equimolar PC. This protective role of PC in both the in vivo and in vitro systems was partially blocked by Indo, although the NSAID had no effect on these properties on its own. CONCLUSIONS: These findings support the hypothesis that PC protects the intestinal mucosa against injurious actions of BS, possibly by forming less toxic mixed micelles. Indo and perhaps other NSAIDs that enter the bile may damage the mucosa, not by a direct action, but by competing for the available protective PC molecules. PMID- 10833494 TI - Cystic fibrosis transmembrane conductance regulator currents in guinea pig pancreatic duct cells: inhibition by bicarbonate ions. AB - BACKGROUND & AIMS: Cystic fibrosis transmembrane conductance regulator (CFTR) Cl( ) channels play an important role in HCO(3)(-) secretion by pancreatic duct cells (PDCs). Our aims were to characterize the CFTR conductance of guinea pig PDCs and to establish whether CFTR is regulated by HCO(3)(-). METHODS: PDCs were isolated from small intralobular and interlobular ducts, and their Cl(- )conductance was studied using the whole-cell patch clamp technique. RESULTS: Activation of a typical CFTR conductance by adenosine 3',5'-cyclic monophosphate (cAMP) was observed in 114 of 204 cells (56%). A larger (10-fold), time- and voltage dependent Cl(-) conductance was activated in 39 of 204 cells (19%). Secretin had a similar effect. Coexpression of both conductances in the same cell was observed, and both conductances had similar anion selectivity and pharmacology. Extracellular HCO(3)(-) caused a dose-dependent inhibition of both currents (K(i), approximately 7 mmol/L), which was independent of intracellular and extracellular pH, and the PCO(2) and CO(3)(2-) content of the bathing solutions. CONCLUSIONS: Two kinetically distinct Cl(-) conductances are activated by cAMP in guinea pig PDCs. Because these conductances are coexpressed and exhibit similar characteristics (anion selectivity, pharmacology, and HCO(3)(-) inhibition), we conclude that CFTR underlies them both. The inhibition of CFTR by HCO(3)(-) has implications for the current model of pancreatic ductal HCO(3)(-) secretion. PMID- 10833496 TI - NF-kappaB/rel/IkappaB: implications in gastrointestinal diseases. PMID- 10833495 TI - Serotonin released from intestinal enterochromaffin cells mediates luminal non cholecystokinin-stimulated pancreatic secretion in rats. AB - BACKGROUND & AIMS: Similar to cholecystokinin (CCK), non-CCK-dependent duodenal factors stimulate vagal mucosal afferent fibers to mediate pancreatic enzyme secretion via a common cholinergic pathway. We tested the hypothesis that 5 hydroxytryptamine (5-HT) released from enterochromaffin (EC) cells plays an important role in the transduction of luminal information to the central nervous system via vagal afferent fibers to mediate pancreatic secretion. METHODS: Pancreatic secretions were examined in conscious rats after intragastric administration of chopped rodent chow in the presence and absence of CCK or 5 HT(3) and 5-HT(2) antagonists. Pancreatic responses to intraduodenal administration of maltose, hyperosmolar NaCl, and light mucosal stroking were examined in rats pretreated with various pharmacological antagonists or after surgical or chemical ablation of vagal and 5-HT neural pathways. RESULTS: Administration of L364, 718 inhibited 54% of pancreatic protein secretion evoked by intragastric administration of rodent chow. L364,714 and ICS 205-930, a 5 HT(3) antagonist, combined produced a 94% inhibition. Vagal afferent rootlet section eliminated pancreatic secretions evoked by intraduodenal stimuli. p Chlorophenylalanine, a 5-HT synthesis inhibitor, but not 5,7-hydroxytryptamine, a 5-HT neurotoxin, also eliminated the pancreatic response to these luminal stimuli. The 5-HT(3) antagonist markedly inhibited pancreatic secretion induced by maltose and hyperosmolar NaCl. 5-HT(2) and 5-HT(3) antagonists combined inhibited the pancreatic response to light stroking of the mucosa. CONCLUSIONS: Luminal factors such as osmolality, disaccharides, and mechanical stimulation stimulated pancreatic secretion via intestinal vagal mucosal afferent fibers. It is likely that 5-HT originating from intestinal EC cells activated 5-HT(3) and 5 HT(2) receptors on vagal afferent fibers to mediate luminal factor-stimulated pancreatic secretion. PMID- 10833497 TI - Presentation of the Julius M. Friedenwald medal to Howard M. Spiro, M.D. PMID- 10833498 TI - American Gastroenterological Association medical position statement: impact of dietary fiber on colon cancer occurrence. American College of Gastroenterology. PMID- 10833499 TI - AGA technical review: impact of dietary fiber on colon cancer occurrence. PMID- 10833500 TI - Bicarbonate secretion and CFTR: continuing the paradigm shift. PMID- 10833501 TI - Vasoactive agents in intrahepatic portal hypertension and fibrogenesis: implications for therapy. PMID- 10833502 TI - Significance of biliary phospholipids for maintenance of the gastrointestinal mucosal barrier and hepatocellular integrity. PMID- 10833503 TI - Decisions, decisions, decisions. PMID- 10833504 TI - Stopping the unstoppable? PMID- 10833506 TI - Therapy of digestive disorders: A companion to sleisenger and Fordtran's gastrointestinal and liver disease PMID- 10833505 TI - Goblet cells make more than just mucus. PMID- 10833507 TI - Regulation of X11L-dependent amyloid precursor protein metabolism by XB51, a novel X11L-binding protein. AB - We isolated a cDNA encoding a novel protein, XB51, that interacts with the amino terminal domain of the neuron-specific X11-like protein (X11L). The protein XB51 inhibited the association of X11L with amyloid precursor protein through a non competitive mechanism and abolished the suppression of beta-amyloid production by X11L. The majority of XB51 is localized around the nucleus and recovered in 3-[(3 cholamidopropyl)dimethylammonio]-1-propanesulfonic acid (CHAPS) buffer-insoluble fraction when XB51 is expressed in cells. Association of XB51 with X11L changed the intracellular distribution of XB51 and resulted in redistribution of XB51 into the CHAPS buffer-soluble fraction. These observations suggest that XB51, together with X11L, plays an important role in the regulatory system of amyloid precursor protein metabolism and beta-amyloid generation. PMID- 10833508 TI - TrkB activation by brain-derived neurotrophic factor inhibits the G protein-gated inward rectifier Kir3 by tyrosine phosphorylation of the channel. AB - G protein-activated inwardly rectifying potassium channels (Kir3) are widely expressed throughout the brain, and regulation of their activity modifies neuronal excitability and synaptic transmission. In this study, we show that the neurotrophin brain-derived neurotrophic factor (BDNF), through activation of TrkB receptors, strongly inhibited the basal activity of Kir3. This inhibition was subunit dependent as functional homomeric channels of either Kir3.1 or Kir3.4 were significantly inhibited, whereas homomeric channels composed of Kir3.2 were insensitive. The general tyrosine kinase inhibitors genistein, Go 6976, and K252a but not the serine/threonine kinase inhibitor staurosporine blocked the BDNF induced inhibition of the channel. BDNF was also found to directly stimulate channel phosphorylation because Kir3.1 immunoprecipitated from BDNF-stimulated cells showed enhanced labeling by anti-phosphotyrosine-specific antibodies. The BDNF effect required specific tyrosine residues in the amino terminus of Kir3.1 and Kir3.4 channels. Mutations of either Tyr-12, Tyr-67, or both in Kir3.1 or mutation of either Tyr-32, Tyr-53, or both of Kir3. 4 channels to phenylalanine significantly blocked the BDNF-induced inhibition. The insensitive Kir3.2 was made sensitive to BDNF by adding a tyrosine (D41Y) and a lysine (P32K) upstream to generate a phosphorylation site motif analogous to that present in Kir3.4. These results suggest that neurotrophin activation of TrkB receptors may physiologically control neuronal excitability by direct tyrosine phosphorylation of the Kir3.1 and Kir3.4 subunits of G protein-gated inwardly rectifying potassium channels. PMID- 10833509 TI - The osteoblast-specific transcription factor Cbfa1 contributes to the expression of osteoprotegerin, a potent inhibitor of osteoclast differentiation and function. AB - Bone formation and resorption are tightly coupled under normal conditions, and the interaction of osteoclast precursors with cells of the osteoblast lineage is a prerequisite for osteoclast formation. Cbfa1 is an osteoblast-specific transcription factor that is essential for osteoblast differentiation and bone formation. At present, it is not known whether Cbfa1 regulates any of the osteoblast-derived factors involved in the bone resorption pathway. Osteoprotegerin (OPG) is an osteoblast-secreted glycoprotein that functions as a potent inhibitor of osteoclast differentiation and bone resorption. Cloning and computer analysis of a 5.9-kilobase human OPG promoter sequence revealed the presence of 12 putative Cbfa1 binding elements (osteoblast-specific element 2 (OSE(2))), suggesting a possible regulation of OPG by Cbfa1. We cloned the promoter upstream of the beta-galactosidase reporter gene (pOPG5. 9betagal) and evaluated whether Cbfa1 could regulate its expression in transient transfection assays. The 5.9-kilobase promoter directed increased levels of reporter gene expression, reminiscent of OPG protein levels in osteoblastic cell lines (BALC and U2OS) as compared with the nonosteoblastic cell line COS1. Cotransfection of a Cbfa1 expression construct along with pOPG5.9betagal reporter construct led to 39-, 7-, and 16-fold increases in beta-galactosidase activity in COS1, BALC, and U2OS cells, respectively. Removal of all the putative OSE(2) elements led to an almost complete loss of transactivation. Mutational analysis demonstrated that the proximal OSE(2) element contributes to a majority of the effects of Cbfa1, and Cbfa1 bound to the proximal element in a sequence-specific manner. Further, overexpression of Cbfa1 led to a 54% increase in OPG protein levels in U2OS cells. These results indicate that Cbfa1 regulates the expression of OPG, thereby further contributing to a molecular link between bone formation and resorption. PMID- 10833510 TI - Pro-sterol carrier protein-2: role of the N-terminal presequence in structure, function, and peroxisomal targeting. AB - Although the 20-amino acid presequence present in 15-kDa pro-sterol carrier protein-2 (pro-SCP-2, the precursor of the mature 13-kDa SCP-2) alters the function of SCP-2 in lipid metabolism, the molecular basis for this effect is unresolved. The presequence dramatically altered SCP-2 structure as determined by circular dichroism, mass spectroscopy, and antibody accessibility such that pro SCP-2 had 3-fold less alpha-helix, 7-fold more beta-structure, 6-fold more reactive C terminus to carboxypeptidase A, 2-fold less binding of anti-SCP-2, and did not enhance sterol transfer from plasma membranes. These differences were not due to protein stability since (i) the same concentration of guanidine hydrochloride was required for 50% unfolding, and (ii) the ligand binding sites displayed the same high affinity (nanomolar K(d) values) in the order: cholesterol straight chain fatty acid > kinked chain fatty acid. Laser scanning confocal microscopy and double immunofluorescence demonstrated that pro-SCP-2 was more efficiently targeted to peroxisomes. Transfection of l-cells or McAR7777 hepatoma cells with cDNA encoding pro-SCP-2 resulted in 45% and 59% of SCP-2, respectively, colocalizing with the peroxisomal marker PMP70. In contrast, l cells transfected with cDNA encoding SCP-2 exhibited 3-fold lower colocalization of SCP-2 with PMP70. In summary, the data suggest for the first time that the 20 amino acid presequence of pro-SCP-2 alters SCP-2 structure to facilitate peroxisomal targeting mediated by the C-terminal SKL peroxisomal targeting sequence. PMID- 10833511 TI - Superoxide regulation of endothelin-converting enzyme. AB - Reactive oxygen species (ROS) act as signaling molecules in the cardiovascular system, regulating cellular proliferation and migration. However, an excess of ROS can damage cells and alter endothelial cell function. We hypothesized that endogenous mechanisms protect the vasculature from excess levels of ROS. We now show that superoxide can inhibit endothelin-converting enzyme activity (ECE) and decrease endothelin-1 synthesis. Superoxide inhibits ECE but hydrogen peroxide and nitric oxide do not. Superoxide inhibits ECE by ejecting zinc from the enzyme, and the addition of exogenous zinc restores enzymatic activity. Superoxide may inhibit other zinc metalloproteinases by a similar mechanism and may thus play an important role in regulating the biology of blood vessels. PMID- 10833512 TI - A 3'-5' exonuclease in human leukemia cells: implications for resistance to 1 beta -D-arabinofuranosylcytosine and 9-beta -D-arabinofuranosyl-2-fluoroadenine 5'-monophosphate. AB - A 3'-5' exonuclease that excises the nucleotide analogs 1-beta-d arabinofuranosylcytosine monophosphate and 9-beta-d-arabinofuranosyl-2 fluoroadenine 5'-monophosphate incorporated at 3' ends of DNA was purified from the nuclei of: 1) primary human chronic lymphocytic leukemia cells, 2) primary and established human acute myeloblastic leukemia cells, and 3) lymphocytes obtained from healthy individuals. The activity of this nuclear exonuclease (exoN) is elevated approximately 6-fold in 1-beta-d-arabinofuranosylcytosine resistant leukemia cells as compared with drug-sensitive cells, and it differs between two healthy individuals and among three leukemia patients. exoN is a 46 kDa monomer, requires 50 mm KCl and 1 mm magnesium for optimal activity, and shows a preference for single-stranded over duplex DNA. Its physical and enzymatic properties indicate that exoN is a previously uncharacterized enzyme whose activity may confer resistance to clinical nucleoside analogs in leukemia cells. PMID- 10833513 TI - Identification of a region of Escherichia coli DnaB required for functional interaction with DnaG at the replication fork. AB - The fundamental activities of the replicative primosomes of Escherichia coli are provided by DnaB, the replication fork DNA helicase, and DnaG, the Okazaki fragment primase. As we have demonstrated previously, DnaG is recruited to the replication fork via a transient protein-protein interaction with DnaB. Here, using site-directed amino acid mutagenesis, we have defined the region on DnaB required for this protein-protein interaction. Mutations in this region of DnaB affect the DnaB-DnaG interaction during both general priming-directed and phiX174 complementary strand DNA synthesis, as well as at replication forks reconstituted in rolling circle DNA replication reactions. The behavior of the purified mutant DnaB proteins in the various replication systems suggests that access to the DnaG binding pocket on DnaB may be restricted at the replication fork. PMID- 10833515 TI - The BCR/ABL tyrosine kinase induces production of reactive oxygen species in hematopoietic cells. AB - The BCR/ABL oncogene causes chronic myelogenous leukemia, a myeloproliferative disorder characterized by clonal expansion of hematopoietic progenitor cells and myeloid cells. It is shown here that transformation of the hematopoietic cell lines Ba/F3, 32Dcl3, and MO7e with BCR/ABL results in an increase in reactive oxygen species (ROS) compared with quiescent, untransformed cells. The increase in ROS was directly due to BCR/ABL because it was blocked by the ABL-specific tyrosine kinase inhibitor STI571. Oxidative stress through ROS is believed to have many biochemical effects, including the potential ability to inhibit protein tyrosine phosphatases (PTPases). To understand the significance of increased production of ROS, a model system was established in which hydrogen peroxide (H(2)O(2)) was added to untransformed cells to mimic the increase in ROS induced constitutively by BCR/ABL. H(2)O(2) substantially reduced total cellular PTPase activity to a degree approximately equivalent to that of pervanadate, a well known PTPase inhibitor. Further, stimulation of untransformed cells with H(2)O(2) or pervanadate increased tyrosine phosphorylation of each of the most prominent known substrates of BCR/ABL, including c-ABL, c-CBL, SHC, and SHP-2. Treatment of the BCR/ABL-expressing cell line MO7/p210 with the reducing agents pyrrolidine dithiocarbamate or N-acetylcysteine reduced the accumulation of ROS and also decreased tyrosine phosphorylation of cellular proteins. Further, treatment of MO7e cells with H(2)O(2) or pervanadate increased the tyrosine kinase activity of c-ABL. Drugs that alter ROS metabolism or reactivate PTPases may antagonize BCR/ABL transformation. PMID- 10833514 TI - Reactive oxygen species generated at mitochondrial complex III stabilize hypoxia inducible factor-1alpha during hypoxia: a mechanism of O2 sensing. AB - During hypoxia, hypoxia-inducible factor-1alpha (HIF-1alpha) is required for induction of a variety of genes including erythropoietin and vascular endothelial growth factor. Hypoxia increases mitochondrial reactive oxygen species (ROS) generation at Complex III, which causes accumulation of HIF-1alpha protein responsible for initiating expression of a luciferase reporter construct under the control of a hypoxic response element. This response is lost in cells depleted of mitochondrial DNA (rho(0) cells). Overexpression of catalase abolishes hypoxic response element-luciferase expression during hypoxia. Exogenous H(2)O(2) stabilizes HIF-1alpha protein during normoxia and activates luciferase expression in wild-type and rho(0) cells. Isolated mitochondria increase ROS generation during hypoxia, as does the bacterium Paracoccus denitrificans. These findings reveal that mitochondria-derived ROS are both required and sufficient to initiate HIF-1alpha stabilization during hypoxia. PMID- 10833516 TI - A novel human gene similar to the protein kinase (PK) coding domain of the large subunit of herpes simplex virus type 2 ribonucleotide reductase (ICP10) codes for a serine-threonine PK and is expressed in melanoma cells. AB - The large subunit of herpes simplex virus type 2 ribonucleotide reductase (ICP10) is a multifunctional protein that contains a serine-threonine protein kinase (PK) activity (Nelson, J. W., Zhu, J. , Smith, C. C., Kulka, M., and Aurelian, L. (1996) J. Biol. Chem. 271, 17021-17027). Phylogenetic analyses indicated that ICP10 PK belongs to a distinct subfamily of growth factor receptor serine threonine PKs that are characterized by their ability to function with a limited number of conserved catalytic motifs (Hunter, J. C. R., Smith, C. C., and Aurelian, L. (1995) Int. J. Onc. 7, 515-522). Here, we report the isolation and characterization of a novel gene, designated H11, that contains an open reading frame of 588 nucleotides, which encodes a protein similar to ICP10 PK. The H11 protein has Mn(2+)-dependent serine-threonine-specific PK activity as determined with a GST-H11 fusion protein and by immununocomplex PK/immunoblotting assays of 293 cells transfected with a H11 eukaryotic expression vector. PK activity is ablated by mutation of Lys(113) within the presumtive catalytic motif II (invariant Lys). 293 cells stably transfected with H11 acquire anchorage independent growth. Endogenous H11 RNA and the H11 phosphoprotein are expressed in melanoma cell lines and primary melanoma tissues at levels higher than in normal melanocytes and in benign nevi. Melanoma cell proliferation is inhibited by treatment with antisense oligonucleotides that inhibit H11 translation, suggesting that H11 expression is associated with cell growth. PMID- 10833517 TI - Developmental stage-specific biosynthesis of glycosylphosphatidylinositol anchors in intraerythrocytic Plasmodium falciparum and its inhibition in a novel manner by mannosamine. AB - Glycosylphosphatidylinositols (GPIs) are the major glycoconjugates in intraerythrocytic stage Plasmodium falciparum. Several functional proteins including merozoite surface protein 1 are anchored to the cell surface by GPI modification, and GPIs are vital to the parasite. Here, we studied the developmental stage-specific biosynthesis of GPIs by intraerythrocytic P. falciparum. The parasite synthesizes GPIs exclusively during the maturation of early trophozoites to late trophozoites but not during the development of rings to early trophozoites or late trophozoites to schizonts and merozoites. Mannosamine, an inhibitor of GPI biosynthesis, inhibits the growth of the parasite specifically at the trophozoite stage, preventing further development to schizonts and causing death. Mannosamine has no effect on the development of either rings to early trophozoites or late trophozoites to schizonts and merozoites. The analysis of GPIs and proteins synthesized by the parasite in the presence of mannosamine demonstrates that the effect is because of the inhibition of GPI biosynthesis. The data also show that mannosamine inhibits GPI biosynthesis by interfering with the addition of mannose to an inositol-acylated GlcN-phosphatidylinositol (PI) intermediate, which is distinctively different from the pattern seen in other organisms. In other systems, mannosamine inhibits GPI biosynthesis by interfering with either the transfer of a mannose residue to the Manalpha1-6Manalpha1-4GlcN-PI intermediate or the formation of ManN-Man-GlcN PI, an aberrant GPI intermediate, which cannot be a substrate for further addition of mannose. Thus, the parasite GPI biosynthetic pathway could be a specific target for antimalarial drug development. PMID- 10833518 TI - Interaction between nucleoside triphosphate phosphohydrolase I and the H4L subunit of the viral RNA polymerase is required for vaccinia virus early gene transcript release. AB - Signal-dependent termination is restricted to early poxvirus genes whose transcription is catalyzed by the virion form of RNA polymerase. Two termination factors have been identified. Vaccinia termination factor/capping enzyme is a multifunctional heterodimer that also catalyzes the first three steps of mRNA cap formation and is an essential intermediate gene transcription initiation factor. Nucleoside triphosphate phosphohydrolase I (NPH I) is a single-stranded DNA dependent ATPase. COOH-terminal deletion mutations of NPH I retain both ATPase and DNA binding activities but are unable either to terminate transcription or to act as dominant negative mutants in vitro. One appealing model posits that the COOH-terminal region of NPH I binds to one or more components in the termination complex. In an attempt to identify NPH I-related protein/protein interactions involved in transcription termination, a series of pull-down experiments were done. Among several vaccinia virus proteins tested, the H4L subunit, unique to the virion form of RNA polymerase, was shown to bind glutathione S-transferase (GST)-NPH I. To further confirm this interaction in virus-infected cells, we constructed recombinant vaccinia virus, vNPHINGST, that expresses GST-tagged NPH I. The H4L subunit of virion RNA polymerase specifically co-purified with GST-NPH I, consistent with a physical interaction. Through the analysis of a series of NH(2)- and COOH-terminal truncation mutations of H4L, the NPH I interaction site was localized to the NH(2)-terminal 195 amino acids of the H4L protein. The H4L binding site on NPH I was mapped to the COOH-terminal region between 457 and 631. Furthermore, COOH-terminal deletion mutations of NPH I failed to bind the NH(2) terminal region of H4L, explaining their inability to support transcription termination. The COOH-terminal end of NPH I was also shown to be required for transcript release activity and for dominant negative inhibition of release. The requirement for an essential interaction between NPH I and H4L provides an explanation for the observed restriction of transcription termination to early viral genes. PMID- 10833519 TI - Persistent yeast single-stranded RNA viruses exist in vivo as genomic RNA.RNA polymerase complexes in 1:1 stoichiometry. AB - Yeast narnavirus 20 S and 23 S RNAs encode RNA-dependent RNA polymerases p91 and p104, respectively, but do not encode coat proteins. Both RNAs form ribonucleoprotein complexes with their cognate polymerases. Here we show that these complexes are not localized in mitochondria, unlike the closely related mitoviruses, which reside in these organelles. Cytoplasmic localization of these polymerases was demonstrated by immunofluorescence and by fluorescence emitted from green fluorescent protein-fused polymerases. These fusion proteins were able to form ribonucleoprotein complexes as did the wild-type polymerases. Fluorescent observations and cell fractionation experiments suggested that the polymerases were stabilized by complex formation with their viral RNA genomes. Immunoprecipitation experiments with anti-green fluorescent protein antibodies demonstrated that a single polymerase molecule binds to a single viral RNA genome in the complex. Moreover, the majority (if not all) of 20 S and 23 S RNA molecules were found to form complexes with their cognate RNA polymerases. Since these viral RNAs were not encapsidated, ribonucleoprotein complex formation with their cognate RNA polymerases appears to be their strategy to survive in the host as persistent viruses. PMID- 10833520 TI - Evidence of an unusually long operator for the fur repressor in the aerobactin promoter of Escherichia coli. AB - Production of the siderophore aerobactin in Escherichia coli is transcriptionally metalloregulated through the iron-dependent binding of the Fur (ferric uptake regulator) to a large region (>100 base pairs) within the cognate promoter in the pColV-K30 plasmid. We show in this article that such an unusually long operator results from the specific addition of degenerate repeats 5'-NAT(A/T)AT-3' and not from a fortuitous occupation of the DNA adjacent to the primary binding sites by an excess of the repressor. Furthermore, the protection pattern revealed by DNase I and hydroxyl radical footprinting reflected a side-by-side oligomerization of the protein along an extended DNA stretch. This type of DNA-protein interactions is more like those observed in some eukaryotic factors and nucleoid-associated proteins than typical of specific prokaryotic regulators. PMID- 10833521 TI - Differential influence of nucleoside analog-resistance mutations K65R and L74V on the overall mutation rate and error specificity of human immunodeficiency virus type 1 reverse transcriptase. AB - Human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) variants with the K65R or L74V substitution display resistance to several nucleoside analogs. An in vitro dNTP exclusion assay revealed an increased fidelity for K65R RT compared with wild-type RT, but little change for L74V RT. When the forward mutation rates were measured via a gap-filling assay, the K65R variant displayed an 8-fold decrease in the overall mutation rate (1.0 x 10(-3) versus 8.6 x 10(-3) for wild-type HIV-1 RT), whereas the rate for the L74V variant was closer to that for wild-type RT (5.0 x 10(-3)). The increase in overall fidelity observed for K65R RT is the largest reported for any drug-resistant HIV-1 RT variant. Nucleotide sequence analysis of lacZalpha mutants generated by variant RTs indicated that K65R RT displays uniform reduction in most types of errors, whereas L74V RT does not. Modeling the substitutions into the x-ray structure of the ternary complex revealed that the major influence of Leu(74) in stabilizing the templating base is unaffected by Val substitution, whereas the K65R substitution appears to increase the stringency of dNTP binding. It is speculated that the increased fidelity of K65R RT is due to an altered interaction with the dNTP substrate. PMID- 10833522 TI - Induction of persistent sodium current by exogenous and endogenous nitric oxide. AB - Most voltage-gated Na(+) channels inactivate almost completely at depolarized membrane potentials, but in some cells a residual Na(+) current is seen that is resistant to inactivation. This persistent Na(+) current can have a profound impact on the electrical behavior of excitable cells, and the regulation of this property could have important biological consequences. However, the biological signaling mechanisms that regulate the persistence of Na(+) channels are not well understood. This study showed that in nerve terminals and ventricular myocytes nitric oxide (NO) reduced the inactivation of Na(+) current. This effect was independent of cGMP, was blocked by N-ethylmaleimide, and could be elicited in cell-free outside-out patches. Thus, a reactive nitrogen species acts directly on the channel or closely associated protein. Persistent Na(+) current could also be induced by endogenous NO generated enzymatically by NO synthase (NOS). Application of ionomycin to raise the intracellular Ca(2+) concentration in myocytes activated NOS. The NO produced in response to ionomycin was detected with an NO-sensitive fluorescent dye. Persistent Na(+) current was enhanced by the same treatment, and NOS inhibitors abolished both the elevation of NO and the induction of persistent Na(+) current. These experiments show that NO is a potential endogenous regulator of persistent Na(+) current under physiological and pathophysiological conditions. PMID- 10833523 TI - Palmitoylation of caveolin-1 is required for cholesterol binding, chaperone complex formation, and rapid transport of cholesterol to caveolae. AB - We previously demonstrated that a caveolin-chaperone complex transports newly synthesized cholesterol from the endoplasmic reticulum through the cytoplasm to caveolae. Caveolin-1 has a 33-amino acid hydrophobic domain and three sites of palmitoylation in proximity to the hydrophobic domain. In the present study, we hypothesized that palmitoylation of caveolin-1 is necessary for binding of cholesterol, formation of a caveolin-chaperone transport complex, and rapid, direct transport of cholesterol to caveolae. To test this hypothesis, four caveolin-1 constructs were generated that substituted an alanine for a cysteine at position 133, 143, or 156 or all three sites (triple mutant). These mutated caveolins and wild type caveolin-1 were stably expressed in the lymphoid cell line, L1210-JF, which does not express caveolin-1, does not form a caveolin chaperone complex, and does not transport newly synthesized cholesterol to caveolae. All of the caveolins were expressed and the proteins localized to plasma membrane caveolae. Wild type caveolin-1 and mutant 133 assembled into complete transport complexes and rapidly (10-20 min) transported cholesterol to caveolae. Caveolin mutants 143 and 156 did not assemble into complete transport complexes, weakly associated with cholesterol, and transported small amounts of cholesterol to caveolae. The triple mutant did not assemble into complete transport complexes and did not associate with cholesterol. We conclude that palmitoylation of caveolin-1 at positions 143 and 156 is required for cholesterol binding and transport complex formation. PMID- 10833524 TI - Overexpression of wild type and mutated human ferritin H-chain in HeLa cells: in vivo role of ferritin ferroxidase activity. AB - Transfectant HeLa cells were generated that expressed human ferritin H-chain wild type and an H-chain mutant with inactivated ferroxidase activity under the control of the tetracycline-responsive promoter (Tet-off). The clones accumulated exogenous ferritins up to levels 14-16-fold over background, half of which were as H-chain homopolymers. This had no evident effect in the mutant ferritin clone, whereas it induced an iron-deficient phenotype in the H-ferritin wild type clone, manifested by approximately 5-fold increase of IRPs activity, approximately 2.5 fold increase of transferrin receptor, approximately 1.8-fold increase in iron transferrin iron uptake, and approximately 50% reduction of labile iron pool. Overexpression of the H-ferritin, but not of the mutant ferritin, strongly reduced cell growth and increased resistance to H(2)O(2) toxicity, effects that were reverted by prolonged incubation in iron-supplemented medium. The results show that in HeLa cells H-ferritin regulates the metabolic iron pool with a mechanism dependent on the functionality of the ferroxidase centers, and this affects, in opposite directions, cellular growth and resistance to oxidative damage. This, and the finding that also in vivo H-chain homopolymers are much less efficient than the H/L heteropolymers in taking up iron, indicate that functional activity of H-ferritin in HeLa cells is that predicted from the in vitro data. PMID- 10833525 TI - Analysis of the inhibition of MyoD activity by ITF-2B and full-length E12/E47. AB - MyoD heterodimerizes with E type factors (E12/E47 and ITF-2A/ITF-2B) and binds E box sequences within promoters of muscle-specific genes. In transient transfection assays, MyoD activates transcription in the presence of ITF-2A but not ITF-2B, which contains a 182-amino acid N-terminal extension. The first 83 amino acids of the inhibitory N terminus of ITF-2B show high sequence homology to the N terminus of full-length E12/E47. Previous studies that showed activation of MyoD by E12 used an artificially N-terminally truncated form. Here we show that the full-length form of E12 inhibits MyoD function. A conserved alpha-helix motif, capable of interacting with the transcriptional machinery, was not essential for inhibition. Furthermore, the fusion of N-terminal ITF-2B sequences or non-inhibiting ITF-2A sequences to truncated E12 was sufficient in converting the activator into an inhibitor. Overexpression of ITF-2B did not inhibit C2C12 myogenesis or affect levels of endogenous muscle gene expression, consistent with the finding that inhibitory E type proteins are present in muscle. Furthermore, we found that MyoD co-transfected with either ITF-2B or ITF-2A converted fibroblasts into myoblasts with the same frequency. Our findings suggest that the ability of E type proteins to inhibit MyoD activity is dependent on the context of the E box. PMID- 10833526 TI - Induction of cytidine to uridine editing on cytoplasmic apolipoprotein B mRNA by overexpressing APOBEC-1. AB - Post-transcriptional editing of apolipoprotein B (apoB) mRNA is regulated in hepatic cells to achieve a steady state proportion of edited and unedited RNA molecules. This activity is catalyzed by APOBEC-1 (apoB mRNA editing catalytic subunit 1) in what has been widely accepted as nuclear event occurring during or after mRNA splicing. Introns impair the efficiency of editing within an adjacent exon in a distance-dependent manner in reporter RNAs. We show here that this inhibition can be overcome by overexpressing APOBEC-1 and that the enhanced editing efficiency on these reporter RNAs occurred after splicing on cytoplasmic transcripts. Given the absolute requirement of auxiliary proteins in apoB mRNA editing, the data suggested that auxiliary proteins were distributed with APOBEC 1 in both the nucleus and cytoplasm of McArdle cells. In fact, immunolocalization of one such auxiliary protein, APOBEC-1 complementation factor (ACF) demonstrated a nuclear and cytoplasmic distribution. We also demonstrate that in the absence of alterations in APOBEC-1 expression, changes in edited apoB RNA induced by ethanol arise through the stimulation of nuclear editing activity. The finding that apoB mRNA editing can occur in the cytoplasm but normally does not suggests that under biological conditions, restricting editing activity to the nucleus must be an important step in regulating the proportion of the edited apoB mRNAs. PMID- 10833527 TI - Families of proteins forming transmembrane channels. AB - Channel-forming proteins/peptides fall into over 100 currently recognized families, most of which are restricted to prokaryotes or eukaryotes, but a few of which are ubiquitous. These proteins fall into three major currently recognized classes: (i) alpha-helix-type channels present in bacterial, archaeal and eukaryotic cytoplasmic and organellar membranes, (ii) beta-barrel-type porins present in the outer membranes of Gram-negative bacterial cells, mitochondria and chloroplasts, and (iii) protein/peptide toxins targeted to the cytoplasmic membranes of cells other than those that synthesize the toxins. High-resolution 3 dimensional structural data are available for representative proteins/peptides of all three of these channel-forming types. Each type exhibits distinctive features that distinguish them from the other channel protein types and from carriers. Structural, functional, and evolutionary aspects of transmembrane channel-formers are discussed. PMID- 10833528 TI - Treatment of cells with detergent activates caspases and induces apoptotic cell death. AB - Due to their amphiphilic properties, detergents readily disrupt cellular membranes and cause rapid cytolysis. In this study we demonstrate that treatment of cells with sublytic concentrations of detergents such as Triton X-100, Nonidet P-40, n-octylglucoside and the bile salt sodium deoxycholate induce typical signs of apoptosis including DNA fragmentation and cleavage of poly(ADP-ribose) polymerase molecules. The detergent concentration required for apoptosis was below the critical micellar concentration. Induction of apoptosis was not restricted to human cells but similarly occurred in a variety of other vertebrate cell lines. Unstimulated peripheral blood mononuclear cells were susceptible to apoptosis induction by detergent suggesting that apoptosis in this circumstance is not mediated by CD95. Cell death was not due to influx of calcium from the medium. Apoptosis was blocked and cytolysis prevented by treatment with peptide inhibitors of caspases. These findings suggest a process of apoptosis that is initiated upon nonspecific alterations at the cell membrane level. Physiologic correlates of this process still have to be defined. PMID- 10833529 TI - K+ secretion in strial marginal cells is stimulated via beta 1-adrenergic receptors but not via beta 2-adrenergic or vasopressin receptors. AB - Pharmacologic tools were used to identify receptors in functional studies by measuring either transepithelial current (I(sc)) in strial marginal cells (SMC) or cAMP production in stria vascularis (SV). Further, receptors were identified in SV as transcripts by cloning and sequencing of reverse-transcriptase polymerase chain reaction (RT-PCR) products. Experiments were performed using tissues isolated from gerbils unless specified otherwise. I(sc) under control conditions was 1090 +/- 21 microA/cm(2) (n = 213) in gerbil SMC and 2001 +/- 95 microA/cm(2) (n = 6) in murine SMC. Direct stimulation of adenylate cyclase with 10(-5) m forskolin but not with 10(-5) m 1,9-dideoxy-forskolin resulted in an increase in the I(sc) by a factor of 1.14 +/- 0.01 (n = 6). The vasopressin receptor agonist 10(-8) m Arg(8)-vasopressin had no significant effect on I(sc) in gerbil and murine SMC. The beta-adrenergic agonists isoproterenol, norepinephrine and epinephrine stimulated I(sc) with an EC(50) of (6 +/- 2) x 10( 7) m (n = 28), (3 +/- 1) x 10(-6) m (n = 40) and (7 +/- 2) x 10(-6) m (n = 38), respectively. Isoproterenol stimulated cAMP production in SV with an EC(50) of (5 +/- 2) x 10(-7) m (n = 8). The beta-antagonist 10(-4) m propanolol completely inhibited 2 x 10(-5) m isoproterenol-induced stimulation of I(sc). The beta antagonists atenolol, ICI118551 and CGP20712A inhibited isoproterenol-induced stimulation of I(sc) with a K(DB) of 1 x 10(-7) m (pK(DB) = 6.96 +/- 0.15, n = 14), 1 x 10(-7) m (pK(DB) = 7. 01 +/- 0.14, n = 15), 2 x 10(-9) m (pK(DB) = 8.73 +/- 0.13, n = 19), respectively. CGP20712A inhibited isoproterenol-induced cAMP production with a K(DB) of 1 x 10(-10) m (pK(DB) = 9.94 +/- 0.55, n = 9). RT-PCR of total RNA isolated from SV using primers specific for the beta(1)-, beta(2)- and beta(3)-adrenergic receptors revealed products of the predicted sizes for the beta(1)- and beta(2)- but not the beta(3)-adrenergic receptor. Sequence analysis confirmed that amplified cDNA fragments encoded gene-specific nucleotide sequences. These results demonstrate that K(+) secretion in SMC is under the control of beta(1)-adrenergic receptors but not beta(2)-adrenergic or vasopressin receptors and that the beta(1)-subtype is the primary beta-adrenergic receptor in SV although SV contains transcripts for both beta(1)- and beta(2)-adrenergic receptors. PMID- 10833530 TI - Evidence for a calcium-sensing receptor in the vascular smooth muscle cells of the spiral modiolar artery. AB - The vascular diameter of the gerbilline spiral modiolar artery has been shown to depend on the presence of extracellular Ca(2+) but it remained unknown whether the smooth muscle cells of this arteriole contain a Ca(2+) sensing receptor (CaSR). The cytosolic Ca(2+) concentration ([Ca(2+)](i)) was monitored as fluo 3 fluorescence and the vascular diameter was measured by video-microscopy in isolated in vitro superfused spiral modiolar arteries. RT-PCR was used to probe for the presence of CaSR transcripts. Increasing the extracellular Ca(2+) concentration ([Ca(2+)](o)) from 1 to 10 mm caused a biphasic increase in [Ca(2+)](i) that was paralleled by a vasoconstriction. The initial rate of this vasoconstriction, 2.01 +/- 0.07 microm/sec (n = 131), was inhibited when cytosolic Ca(2+) stores were presumably depleted with thapsigargin (IC(50) = 3 x 10(-9) m, n = 26) or ryanodine (IC(50) = 4 x 10(-8) m, n = 25) or when PLC was inhibited by 10(-6) m U73122 (n = 8). The initial rate of this constriction was not affected by the L-type Ca(2+) channel blocker 10(-6) m nifedipine (n = 5), by 10(-6) m U73343 (n = 6), which is the inactive analogue of U73122, by the T-type Ca(2+) channel blocker 10(-6) Gd(3+) (n = 6) or the Na(+)/Ca(2+) exchanger blocker 10(-4) m Ni(2+) (n = 5). The agonist rank potency order was Gd(3+) > Ni(2+) > Ca(2+) >> neomycin = Mg(2+). Analysis of RNA isolated from the SMA revealed a RT-PCR product of the appropriate size for the CaSR (448 bp). Sequence analysis of the amplified cDNA fragment revealed a 94-96% amino acid identity compared to other CaSRs. These results demonstrate that the spiral modiolar artery contains a CaSR, which is most likely located in the vascular smooth muscle cells. PMID- 10833531 TI - Bidirectional transepithelial water transport: chloride-dependent mechanisms. AB - We hypothesized that inhibition and activation of basolateral to luminal chloride transport mechanisms were associated with respective decreases and increases in basolateral to luminal water fluxes. The luminal to basolateral (J(W)(L-->B)) and basolateral to luminal (J(W)(B-->L)) water fluxes across ovine tracheal epithelia were measured simultaneously. The mean J(W)(L-->B) (6.5 microl/min/cm(2)) was larger than J(W)(B-->L) (6.1 microl/min/cm(2)). Furosemide reduced J(W)(B-->L) from 6.0 to 5.6 microl/min/cm(2). Diphenylamine-2-carboxylate (DPC) reduced J(W)(B-->L) from 7.9 to 7. 3 microl/min/cm(2) and reduced the membrane potential difference by 38%. Furosemide together with DPC decreased J(W)(L-->B) by 30% and J(W)(B-->L) by 15%. Norepinephrine increased J(W)(B-->L) from 4.9 to 6.0 microl/min/cm(2). Neuropeptide Y in the presence of norepinephrine decreased J(W)(L-->B) (6.4 to 5.2 microl/min/cm(2)) and returned J(W)(B-->L) to its baseline value. Vasopressin increased J(W)(B-->L) from 4.1 to 5.1 microl/min/cm(2). Endothelin-1 induced a simultaneous increase in J(W)(B-->L) (7.0 to 7.7 microl/min/cm(2)) and decrease in J(W)(L-->B) (7.4 to 6.4 microl/min/cm(2)); and decreased the membrane resistance. These data indicate that in tracheal epithelia under homeostatic conditions J(W)(B-->L) has a approximately 15% actively coupled component. Consistent with our hypothesis, inhibition and receptor-induced stimulation of chloride effluxes were associated with decreases and increases in J(W)(B-->L), respectively. However, as inhibition of transcellular chloride transport always decreased J(W)(L-->B) more than J(W)(B ->L), reducing transepithelial chloride transport did not result in less water being transported into the airway lumen. PMID- 10833532 TI - Skeletal muscle ryanodine receptor channels are activated by the fungal metabolite, gliotoxin. AB - Interactions between the reactive disulfide fungal metabolite, gliotoxin (GTX), and rabbit skeletal ryanodine receptor (RyR) calcium release channels have been examined. RyRs in terminal cisternae vesicles formed a covalent complex with 100 microm (35)S-GTX, which was reversed by 1 mm dithiothreitol (DTT) or 1 mm glutathione. GTX (80-240 microm), added to either cytoplasmic (cis) or luminal (trans) solutions, increased the rate of Ca(2+) release from SR vesicles and the frequency of opening of single RyR channels in lipid bilayers. Channel activation was reversed upon addition of 2 mm DTT to the cis solution, showing that the activation was due to an oxidation reaction (2 mm DTT added to the cis solution in the absence of GTX did not affect RyR activity). Furthermore, RyRs were not activated by trans GTX if the cis chamber contained DTT, suggesting that GTX oxidized a site in or near the membrane. In contrast to cis DTT, 2 mm DTT in the trans solution increased RyR activity when added either alone or with 200 microm trans GTX. The results suggest that (i) GTX increases RyR channel activity by oxidizing cysteine residues that are close to the membrane and located on RyR, or associated proteins, and (ii) a disulfide bridge or nitrosothiol, accessible only from the luminal solution, normally suppresses RyR channel activity. Some of the actions of GTX in altering Ca(2+) homeostatsis might depend on its modification of RyR calcium channels. PMID- 10833533 TI - Furosemide stimulates K transport in HCD57 erythroid cells. AB - We examined the influence of serum and furosemide on K movement and cell volume in HCD57 cells, a murine erythroleukemia cell line, which require erythropoietin (EPO) for survival. We found that maintenance of cell volume depends on the concentration of serum in the culture medium. In isotonic medium containing 20% serum, HCD57 cells maintain their steady-state volume. In contrast, the cells shrink progressively as medium serum content is reduced. In serum-free medium, raising external K to 75 mm prevents cell shrinkage and a further increase in K to 145 mm results in swelling, revealing a role for K permeability in the regulation of cell volume. Of particular interest has been a serendipitous finding with furosemide. Below an external K concentration of 2.1 +/- 0.3 mm in medium containing 2% serum, furosemide inhibits K uptake, probably stemming from its well known inhibitory action on KCl cotransport. However, above that K concentration, furosemide stimulates K uptake in a dose-dependent manner. Moreover, furosemide potentiates cell shrinkage induced by serum withdrawal. These findings suggest that the transport machinery mediating cellular shrinkage, once primed by serum depletion, becomes receptive to a second stimulus. PMID- 10833534 TI - Commitment to research PMID- 10833535 TI - Hypertension management in clinical practice. PMID- 10833536 TI - The thoughtful baker. PMID- 10833537 TI - The importance of knowing about the incidence of acute myocardial infarction. PMID- 10833538 TI - Trends in the incidence of acute myocardial infarction between 1984 and 1993 - The Halifax County MONICA Project. AB - BACKGROUND: According to vital statistics data for Halifax County, between 1984 and 1993 the annual mortality rate decreased for ischemic heart disease and myocardial infarction (MI). OBJECTIVES: To estimate the change in MI mortality, applying standardized diagnostic criteria; to determine whether decreased case fatality or decreased MI event rate, or both, caused decreased mortality; and to determine the contribution of MI incidence rate to altered event rate. PATIENTS AND METHODS: All persons in the study area aged 25 to 74 years and admitted to hospital or dying outside hospital with suspected acute coronary syndromes were registered prospectively. Demographic, health history and clinical data were extracted from medical records or collected from medical examiner reports, next of-kin interviews or family physicians. Definite or possible MI was diagnosed according to World Health Organization MONItoring of trends and determinants in CArdiovascular disease (MONICA) criteria. Trends in age- and sex-standardized rates were estimated by using log-linear regression analysis. RESULTS: Of 4283 patients admitted to hospital for MI, 23.9% died within 28 days; 1401 patients who had suffered an MI died before admission to hospital. MI mortality decreased annually by 3.9% (95% CI 1.9 to 5.8); two-thirds of the decline was due to MI event rates (2.6%; CI 1.3 to 3.8) and one-third to a decrease in 28-day case fatality (1.3%; CI 0.2 to 2. 3). A decrease in MI incidence rate (3.2%; CI 1.7 to 4.8), rather than a decline in MI recurrence rate (1.4%; CI 0.7 to -3.5), was the major reason for the declining event rate. CONCLUSIONS: A decrease in the incidence of MI, possibly due to primary prevention, had a major impact on the declining MI mortality. Decreased in-hospital MI fatality, possibly due to improved treatment, was responsible for the decline in case fatality. PMID- 10833539 TI - Trends in the in-hospital treatment of acute myocardial infarction between 1984 and 1993 - The Halifax County MONICA Project. AB - BACKGROUND: The Halifax County MONItoring of trends and determinants in CArdiovascular disease (MONICA) Project found that between 1984 and 1988, the proportion of myocardial infarctions (MIs) that were fatal within 28 days remained constant, but declined between 1989 and 1993. The objective was to investigate association among case fatality, treatment and case severity of MI in hospitalized patients. PATIENTS AND METHODS: The MONICA MI register contains data on demographics, health history, in-hospital investigations, interventions and treatment, and vital status at 28 days after onset of symptoms for all MIs occurring in residents of Halifax County, aged 25 to 74 years. Logistic regression analysis was used to estimate trends in the use of cardioactive drugs and revascularization procedures. A case severity score was developed from patient characteristics at time of admission. Case fatality was calculated as the proportion of MIs that were fatal within 28 days. RESULTS: Between 1984 and 1988, a large increase (OR 1.3) occurred in the use of angiotensin-converting enzyme (ACE) inhibitors, acetylsalicylic acid (ASA), thrombolysis and percutaneous transluminal coronary angioplasty (PTCA); a minor increase occurred in use of calcium channel blockers (OR=1.29, 99% CI 1.19 to 1.40); beta-blocker use decreased; case fatality remained constant and case severity score increased. From 1989 to 1993, ACE inhibitor use increased (OR=1.4, 99% CI 1.27 to 1.55); minor increases occurred in use of ASA and beta-blockers, and in PTCA and coronary artery bypass grafting; case severity did not change and case fatality decreased. CONCLUSIONS: While use of beneficial treatment increased between 1984 and 1988, MI case fatality did not decrease, probably because case severity increased. Between 1989 and 1993, case severity remained constant, and the further increase in the use of beneficial therapy was associated with a decline in case fatality. PMID- 10833540 TI - Pericardial pressure in experimental chronic heart failure. AB - BACKGROUND: In the normal heart, pericardial pressure is greater than previously believed. OBJECTIVES: To explore the contribution of pericardial constraint to the elevated left ventricular (LV) end-diastolic pressure in chronic heart failure (CHF). ANIMALS AND METHODS: Pericardial pressure was measured directly in 11 dogs with CHF. Seven dogs were instrumented with LV and right ventricular micromanometers and epicardial pacing leads, and paced at 240 to 260 beats/min for four to seven weeks. After the development of CHF, a left thoracotomy was performed and a flat pericardial balloon was positioned over the LV free wall through a slit in the pericardium. RESULTS: LV end-diastolic pressure was 31+/-9 mmHg, and pericardial pressure only 7+/-2 mmHg. Nitroglycerin in six dogs decreased LV end-diastolic pressure from 33+/-8 to 28+/-7 and pericardial pressure from 7+/-2 to 6+/-3 mmHg (both P<0.05). Calculated transmural LV end diastolic pressure also decreased (26+/-8 to 22+/-7 mmHg, P<0.05). Volume loading in five dogs increased LV end-diastolic pressure from 29+/-8 to 42+/-10 mmHg (P<0.05), pericardial pressure from 6+/-3 to 12+/-6 mmHg (not significant) and transmural LV end-diastolic pressure from 23+/-7 to 30+/-7 mmHg (not significant). When the pericardium was opened in three dogs, the LV end-diastolic pressure decreased by 5 mmHg. Four previously uninstrumented dogs were studied to exclude the effects of epicardial scarring; LV end-diastolic pressure was 42+/-6 mmHg and pericardial pressure was 10+/-6 mmHg. CONCLUSION: Pericardial constraint, a prerequisite for pericardially mediated ventricular interaction, was not present to the same extent in this model of CHF as in acute models, probably reflecting the importance of pericardial remodelling. PMID- 10833541 TI - Opposite effects of amlodipine and enalapril on infarct collagen and remodelling during healing after reperfused myocardial infarction. AB - OBJECTIVES: To compare the effects of the calcium channel blocker amlodipine versus the angiotensin-converting enzyme inhibitor enalapril with or without reperfusion on infarct collagen and remodelling during healing after anterior myocardial infarction (MI). ANIMALS AND METHODS: In vivo left ventricular (LV) remodelling and function (by quantitative echocardiography) and hemodynamics were measured over six weeks in dogs that were randomized 24 h after reperfusion (2 h after anterior MI) or no reperfusion to oral amlodipine (5 mg bid, n=6), enalapril (5 mg bid, n=6), placebo (bid, n=6) or sham surgery (n=6) for six weeks. Ex vivo infarct size, infarct collagen (hydroxyproline), collagen volume fraction and LV topography were measured at six weeks. RESULTS: Compared with placebo controls without reperfusion over six weeks in vivo, enalapril or amlodipine with or without reperfusion produced LV unloading and preserved volumes, shape and function, but enalapril limited LV hypertrophy more than amlodipine. However, compared with no reperfusion, amlodipine preserved infarct wall thickness and shape while enalapril decreased infarct wall thickness and increased the shape index. Ex vivo at six weeks, scar size as a percentage of risk was similar in the MI groups. Importantly, enalapril decreased infarct collagen already lowered by reperfusion, while amlodipine preserved infarct collagen after reperfusion and increased collagen volume fraction in spared myocardium. CONCLUSIONS: Preservation of infarct collagen limits infarct remodelling during healing after reperfused MI and preserves LV shape. Amlodipine and enalapril exert opposite effects on infarct collagen and remodelling after reperfused MI. PMID- 10833542 TI - Implantable ventricular assist devices: is it time to introduce them in Canada? AB - BACKGROUND: Implantable left ventricular assist devices (LVAD) are increasingly used in Europe and the United States. Any decision to use them in Canada requires estimates of their clinical value and costs. MATERIALS AND METHODS: No randomized controlled trials are available. Clinical value and costs, concerning principally the HeartMate and Novacor devices, were estimated based on reports of uncontrolled case series obtained through MEDLINE (1993 to 1999), review articles, three technology assessments and data supplied by the manufacturers. RESULTS AND DISCUSSION: Reasonably trouble-free device function can be expected for three to four years. The principal application is as a bridge to transplantation. Rarely, the heart recovers without transplantation. Use as 'permanent' support of the failing heart is still contentious. Approximately 70% of patients with an implanted LVAD survive until recovery or transplantation. Complications are hemorrhage, principally postoperative, 20% to 44%; thromboembolism, ranging from 5% to 15% for the HeartMate to 12% to 37% for Novacor; and significant infection, 50%. Quality of life is slightly inferior to that of patients with transplanted hearts. The direct cost to the health care system of installation is approximately $138,000. As a bridge to transplantation, the estimated cost effectiveness of elective interventions is $91,000 to $126,000 per life-year saved ($117,000 to $186,000, discounted at 5%), and as a permanent alternative to transplantation, the cost per life-year is $52,000 to $60,000 ($50, 000 to $58,000, discounted at 5%), according to circumstances. As a bridge to 50 transplantations per year, the approximate annual cost would be $7 to $13 million (exclusive of transplantation costs). As 'permanent' support for 7000 patients per year, the approximate cost would be $2,661 million per year. CONCLUSIONS: Limited application in a limited number of centres with collection of all data is justifiable at this stage. PMID- 10833543 TI - The enigma of recent onset dilated cardiomyopathy: what cause, what consequence, what control? AB - Acute onset dilated cardiomyopathy is a very common presentation encountered by cardiologists in clinical practice, but little is known about the etiology, pathophysiology, definitive diagnosis and management of this syndrome. The vast majority of cases are considered as idiopathic cardiomyopathy, or primary or secondary myocarditis, although rarely definitive diagnoses are obtained after extensive diagnostic procedures. Two scenarios are presented as a basis for a discussion of this challenging syndrome. PMID- 10833544 TI - The impact of depression on the course and outcome of coronary artery disease: review for cardiologists. AB - Epidemiological research over the past decade indicates that major depression and depressive symptomatology are common among patients with coronary artery disease (CAD) and postmyocardial infarction. Major depression is a serious, debilitating comorbid disorder that can significantly increase risk for and complicate recovery from cardiac events (eg, myocardial infarction). Unfortunately, major depression is rarely detected or treated in the cardiology setting. However, it is a disorder that can be successfully and safely treated in the majority of cases. Although the mechanisms that may be mediating the depression-CAD link are less well understood, there are several plausible mechanisms by which depression may influence the course and outcome of CAD. This review examines the literature linking major depression and depressive symptomatology to CAD course and outcome, and makes recommendations for improving assessment and treatment of depression in the cardiology setting. PMID- 10833545 TI - Mesalamine-induced chest pain: a case report. AB - Cardiac side effects of mesalamine are uncommon. A young man with ulcerative colitis who developed recurrent chest pain and electrocardiographic changes while on mesalamine is presented. Various causes of mesalamine-induced chest pain are discussed. PMID- 10833546 TI - A complicated case of pericardial hydatid cyst manifesting as constrictive pericarditis. AB - Although cardiac involvement with hydatid cyst is quite rare as a major complication, constrictive pericarditis is even less common. A 55-year-old man is presented in whom a hydatid cyst located in the right cardiophrenic angle anterior to the right ventricle ruptured into the pericardial sac, resulting in constrictive pericarditis. PMID- 10833548 TI - Editorial PMID- 10833547 TI - A look back at 1999 and ahead to 2001 PMID- 10833549 TI - Beware somatization. PMID- 10833550 TI - Wind-up and the NMDA receptor complex from a clinical perspective. AB - More rational treatment of chronic pain depends on increased understanding of the pathophysiological mechanisms underlying the various symptoms and characteristics of chronic pain. Central sensitization of pain represents an important pathophysiological mechanism that has received great attention in recent years. The experimental models used to explore mechanisms of central sensitization include the study of wind-up in animals and temporal summation of pain in humans. Wind-up was described more than 30 years ago as progressively increasing activity in dorsal horn cells following repetitive activation of primary afferent C fibres. In humans, temporal summation of repeated painful stimuli has been regarded as a psychophysical correlate of wind-up. This review focuses on the relationship between wind-up, temporal summation and central sensitization. In particular, the role of NMDA receptor mechanisms in the modulation of wind up/temporal summation is discussed. The data presented here indicate that the study of wind-up and temporal summation has given information about some of the complex mechanisms underlying central sensitization. Both wind-up and temporal summation appear to be dependent on NMDA receptor activation. The results of clinical trials in patients with chronic pain suggest that the NMDA receptor may represent a new target for modulation of abnormal temporal summation of pain, as well as other characteristics of chronic pain. PMID- 10833551 TI - Commentary: the peripheral mechanisms of abnormal temporal summation. PMID- 10833552 TI - Neuronal inhibitory effects of methadone are predominantly opioid receptor mediated in the rat spinal cord in vivo. AB - This study aims to assess whether the antinociceptive actions of methadone are mediated solely through opioid mechanisms, or whether its reported affinity for NMDA receptors has physiological relevance in vivo. Methadone is a mu-opioid receptor agonist reported to relieve pain unresponsive to other opioids. It is a racemic mixture comprising d- and l-optical isomers; the d-isomer has a lower affinity for opioid receptors, and both also exhibit NMDA receptor binding, likely to indicate antagonist activity. d -Methadone is antinociceptive in behavioural studies via non-opioid mechanisms, which could include functional NMDA receptor-blocking activity. Here we investigate the ability of d - and dl methadone to inhibit noxious and innocuous electrically-evoked responses of dorsal horn neurones in the anaesthetized rat. Racemic methadone (5, 25, 50, 250 microg) applied spinally, dose-relatedly inhibited the C-fibre evoked response, input and wind-up of the neurones, with a profile resembling that of morphine. d Methadone (5, 25, 50, 250, 500 microg) was also inhibitory, although less potent by a factor of between 13 and 48 depending on the neuronal measure; its profile of inhibition resembled that of the racemic mixture rather than an NMDA receptor antagonist. Both compounds had minimal effects on Abeta-fibre-evoked activity. The inhibitory effects of both d - and dl -methadone on noxious-evoked activity were naloxone reversible. The naloxone reversibility of d -methadone inhibitions is best interpreted as indicative of a purely opioid mechanism of action. However, the ability of naloxone to reverse the effects of d -methadone may also reflect a degree of synergy between weak NMDA antagonist and opioid agonist activity. PMID- 10833553 TI - Citalopram in patients with fibromyalgia--a randomized, double-blind, placebo controlled study. AB - The effect of the selective serotonin reuptake inhibitor citalopram was studied in a randomized, double-blind, placebo-controlled, 4-month trial in patients with the fibromyalgia syndrome (FMS) who all fulfilled the American College of Rheumatology criteria. The citalopram doses varied between 20-40 mg daily. Forty female patients, 21 patients in the citalopram and 19 in the placebo group, participated. Assessment of pain, depressive symptoms and physical functioning were made using Visual Analogue Scales (VAS), the Montgomery Asberg Depression Rating Scale (MADRS) and the Fibrositis Impact Questionnaire (FIQ). In the global judgement of improvement, no significant changes were found between the citalopram and placebo groups as concerns pain or well-being, either in the Intention to Treat (ITT) analysis or in the completer analysis. However, among the completers, it was a tendency that more patients in the citalopram group (52.9%) were improved as compared to the placebo group (22.2%) concerning well being. Furthermore, the results indicated that treatment with citalopram had a significant effect on pain on the VAS after 2 months of treatment compared to baseline. After 4 months, however, the effect had diminished. Measured with the FIQ, significant differences in the pain ratings were seen at the end of the trial. Significant effects on the depressive symptomatology measured by means of the MADRS were seen already after 1 month of treatment and were increasing further at the end of the trial, when a significant difference between the groups was also found. PMID- 10833554 TI - The emotional stroop task and chronic pain: what is threatening for chronic pain sufferers? AB - Using a computer version of the emotional stroop task, it was investigated whether chronic pain patients display an involuntary attentional shift towards pain-related information (sensory, affective pain words and injury related words). Multiple regression analyses were used to investigate which pain and psychosocial variables (pain severity, pain-related fear, pain catastrophizing and negative affect) were predictive of attentional bias. Results indicated: (1) that there was an attentional bias towards the sensory pain words; and (2) that current pain intensity was predictive of the effect. No other attentional effects were found. The results are discussed in terms of possible reasons for the difficulty of demonstrating attentional bias in chronic pain patients. PMID- 10833555 TI - The DSM-IV nosology of chronic pain: a comparison of pain disorder and multiple somatization syndrome. AB - This study evaluates the classification of pain from the perspective of the DSM IV system. Of 60 in-patients with long-standing and disabling pain syndromes, 29 with pain disorder (PD) and 31 with pain as part of a multiple somatization syndrome (MSS) were compared before and after a structured cognitive-behavioral treatment. It was hypothesized that MSS patients show more psychological distress, are more severely disabled, and respond less to the treatment. Both groups were similar with respect to sociodemographic status, history of pain symptomatology and comorbidity with DSM-IV mental disorders. The results show that MSS patients had higher levels of affective and sensoric pain sensations as well as more pain-related disabilities. They were also less successful during treatment to reduce their pain-related depression and anxiety. Psychosocial functioning was improved only by PD patients, but remained almost unchanged in the MSS group. However, there were no group differences concerning general depression and hypochondriasis, dysfunctional attitudes towards body and health, and use of pain coping strategies. It is concluded that the DSM-IV distinction between 'pure' pain disorder and syndromes involving pain plus multiple somatoform symptoms cannot generally be confirmed, but further studies of validation are needed. PMID- 10833556 TI - The influence of pain intensity on somatosensory perception in patients suffering from subacute/chronic lateral epicondylalgia. AB - A confounding factor in the analysis of chronic pain patients is the finding of signs of somatosensory disturbances not only in neuropathic pain patients but also in a subgroup of patients with musculoskeletal pain. The purpose was to investigate if patients suffering from subacute/chronic lateral epicondylalgia demonstrated altered sensibility, and if this was affected by pain intensity. At the start of the experiment, quantitative sensory testing (QST) (thermal, pressure pain, touch) was performed in the local pain area and in the area of pain referral. QST was repeated following pain provocation (weight lifting). A local anaesthetic was then injected into the lateral epicondyle and QST was repeated in the area of pain referral. The contralateral arm was assessed, treated and injected in the same way. At the baseline assessment there was no difference in sensibility between sides, with the exception of a significantly lowered threshold to noxious heat (p<0.04) in the area of pain referral, present during the whole experiment. In the affected arm only, weight lifting resulted in significantly increased pain intensity in the local (p<0.01) and referred (p<0.01) pain areas, respectively. Repeated muscle contractions resulted in altered somatosensory functions in both the affected arm and the unaffected arm, consequently not dependent on ongoing pain in the assessed area. Tactile perception thresholds increased significantly following pain provocation in the area of pain referral (p<0.04) only and normalized following injection of local anaesthetic (p<0.02), indicating that the sensitivity to light touch was altered by the nociceptive input from the affected arm. PMID- 10833557 TI - Injection of hypertonic saline into musculus infraspinatus resulted in referred pain and sensory disturbances in the ipsilateral upper arm. AB - A confounding factor in the analysis of chronic pain patients is the finding of somatosensory disturbances not only in neuropathic pain patients, but also in a subgroup of patients with musculoskeletal pain. The purpose of the study was to examine if referred pain, induced by intramuscular injections of hypertonic saline (5% NaCl) into the left musculus infraspinatus, resulted in somatosensory alterations. Thermal sensitivity, pressure pain sensitivity, as well as low threshold mechanoreceptive function, were assessed in the referred pain area and the homologous contralateral site before, during and following the injections. In 10 out of 12 subjects the procedure induced only referred pain localized in the dorsolateral part of the ipsilateral proximal upper arm. In this referred pain area there was a significantly decreased sensitivity to light touch, as tested with von Frey filaments, during the pain period and the post-injection period compared to the contralateral side (p<0.004 and p<0.009, respectively). A trend for thermal hypoaesthesia, which was only demonstrable in the sum of warm and cold thresholds, was found in the referred pain area, but not contralaterally, during the pain period compared to the pre-injection period. Significantly increased sensitivity to threshold and suprathreshold heat pain was found bilaterally during post-injection assessments (p<0.02 and p<0.006, respectively). There were no statistically significant changes in sensitivity to innocuous thermal stimuli when assessing the two percepts separately, or to pressure pain or brush-evoked touch. In conclusion, intramuscular injections of hypertonic saline resulted in referred pain and tactile hypoaesthesia in the referred pain area. PMID- 10833558 TI - Descending facilitatory modulation of a behavioral nociceptive response by stimulation in the adult rat anterior cingulate cortex. AB - It is well documented that the descending endogenous analgesia system, including the periaqueductal gray (PAG) and the rostral ventral medulla (RVM), play an important role in modulation of nociceptive transmission and morphine- and cannabinoid-produced analgesia. Neurons in the PAG receive inputs from different nuclei of higher structures, including the anterior cingulate cortex (ACC). However, it is unclear if stimulation of neurons in the ACC modulates spinal nociceptive transmission. The present study has examined the effects of electrical stimulation and chemical activation of metabotropic glutamate receptors (mGluRs) in the ACC on a spinal nociceptive tail-flick (TF) reflex induced by noxious heating. Activation of the ACC at high intensities (up to 500 microA) of electrical stimulation did not produce any antinociceptive effect. Instead, at most sites within the ACC (n = 36 of 41 sites), electrical stimulation produced significant facilitation of the TF reflex (i.e. decreases in TF latency). Chemical activation of mGluRs within the ACC also produced a facilitatory effect. Descending facilitation from the ACC apparently relays at the RVM. Electrical stimulation in the RVM produces a biphasic modulatory effect, showing facilitation at low intensities and inhibition at higher intensities. The present study provides evidence that activation of mGluRs within the ACC can facilitate spinal nociception. PMID- 10833559 TI - The anticonvulsant remacemide and its metabolite AR-R12495AA attenuate spinal synaptic transmission and carrageenan-induced inflammation in the young rat. AB - The effects of the anticonvulsants remacemide [(+/-)-2-amino-N-(1-methyl-1,2 diphenylethyl)-acetamide hydrochloride] and its des -glycinated metabolite AR R12495AA [(+/-)-1-methyl-1,2-diphenylethylamine- monohydrochloride] on primary afferent-induced synaptic transmission and frequency-dependent summation of synaptic potentials were assessed in the young rat spinal cord in vitro. Behavioural studies in the rat determined the effects of these anticonvulsant compounds in the carrageenan model of inflammation. Recordings of the extracellular dorsal root-evoked ventral root potential (DR-VRP) revealed a significant reduction of the duration and t(1)-(2)decay of the long latency, slow DR-VRP by remacemide (50 and 100 microM) and AR-R12495AA (25, 50 and 100 mM). The short-latency, fast monosynaptic DR-VRP peak was reduced by only the highest concentration of AR-R12495AA (100 microM). In intracellular dorsal root-evoked excitatory postsynaptic potentials (DR-EPSPs) of single ventral horn neurons, AR R12495AA (100 microM) attenuated the time course of the long-latency (slow) EPSP. Frequency-dependent (0.5-2.0 Hz) summation of dorsal root-evoked synaptic events (recorded extracellularly as the cumulative ventral root depolarization (CVRD), and intracellularly as wind-up) was attenuated by remacemide (100 microM) and AR R12495AA (50 and 100 microM). Pre-treatment with intra-peritoneal injection of 75 mg/kg of remacemide or AR-R12495AA caused a significant reduction of carrageenan induced mechanical hyperalgesia and oedema. These electrophysiological and behavioural data provide evidence that remacemide and AR-R12495AA may also possess analgesic and anti-inflammatory activity. PMID- 10833560 TI - The effects of exogenous analgesia in a patient with borderline personality disorder (BPD) and severe self-injurious behaviour. AB - In order to analyse the effects of potent exogenous analgesia with opioids in borderline personality disorder (BPD), we present a case report in which the application of morphine abolished the perception of pain during self-injury and intensified self-injurious activities. On the basis of our observations, we concluded that the use of potent analgesics might aggravate psychopathology in BPD. PMID- 10833561 TI - Lumbar catheterization of the subarachnoid space with a 32-gauge polyurethane catheter in the rat. AB - Chronic catheterization of the subarachnoid space of rats is an important tool for intrathecal drug delivery in pharmacologic investigations of pain. We describe a technique using direct lumbar insertion of a small 32-gauge polyurethane (PU) catheter without extensive surgery. Location of the catheter was confirmed with 2% lidocaine injection 1 day later, and methylene blue injection after 7-14 days. This method improved recovery of the rat after catheter implantation and reduced neurologic complications. PMID- 10833562 TI - Dystonia: recent advances. AB - Dystonia may be primary or symptomatic. Most, if not all, primary torsion dystonias are genetic diseases and manifest as 'pure dystonia', without consistent biochemical or neuropathological changes. Symptomatic dystonias may be (a) secondary to drugs or other environmental factors, (b) part of a 'dystonia plus' syndrome or (c) part of several heredodegenerative diseases. In the last few years, there have been rapid advances in the genetic classification of primary torsion dystonia. The gene for one form (DYT1dystonia), particularly common in Ashkenazi Jews, has been isolated. In this review, I present a basic clinical overview of dystonia and focus on the recent advances in molecular genetics of primary torsion dystonia (PTD). Treatment of dystonia is a large subject, worthy of a review in itself, and is not covered here. Several of the paroxysmal movement disorders may manifest with dystonia, but these are usually considered separately, as I have done in this review. Copyright 1999 Harcourt Publishers Ltd. PMID- 10833563 TI - SPECT abnormalities in Landau-Kleffner syndrome. AB - Five right-handed children with acquired aphasia elipepsy syndrome (Landau Kleffner, LKS), were investigated with 99(m)TcHMPAO single photon emission computed tomography (SPECT) and the results were correlated with their EEGs and clinical history. The childrens' ages ranged from 2 to 5 years and the aphasia had been present for 6 to over 12 months. No clinical seizure had ever been onserved in the younger two children and their waking EEGs showed infrequent central spikes. Both children had areas of low intensity on SPECT, involving the left temporal lobe in one and the right temporal lobe in the other, which has also been reported in children with congenital dysphasia who have normal EEGs. The three older children presented with frequent generalized seizures, with the aphasia occurring 3-6 months later. The SPECT scans in these children were performed either in the ictal state, or when electrographic seizure activity was very frequent on EEG. All three children had hyperintense foci on SPECT involving the left posterior temporal region corresponding to Wernickes area. We conclude that LKS may be initially a unilateral seizure disorder of Wernickes area, with EEG discharges in the contralateral hemisphere representing propagation from the unilateral focus. Copyright 1999 Harcourt Publishers Ltd. PMID- 10833564 TI - Central motor conduction time in Parkinson's disease. AB - Central motor conduction time (CMCT) was analysed in 11 patients with Parkinson's disease and 10 age matched healthy controls. CMCT was measured during muscle relaxation, muscle contraction and in patients with Parkinson's disease before and 1 h after intravenous infusion of L-dopa (0.25%, 20 ml). Although the CMCT during muscle contraction did not differ significantly between Parkinson's disease and healthy controls, the CMCT during muscle relaxation was significantly shorter in Parkinson's disease than in healthy controls. The CMCT during muscle relaxation in Parkinson's disease was significantly longer after intravenous infusion of L-dopa than before, but the CMCT during muscle contraction did not differ significantly before and after intravenous infusion of L-dopa. The CMCT during muscle contraction was significantly shorter than the CMCT during muscle relaxation in healthy controls. However, in Parkinson's disease, the CMCT did not differ significantly between muscle relaxation and contraction. We consider that the central motor conduction pathway during muscle relaxation in Parkinson's disease is modified to be similar to that during muscle contraction in healthy subjects. Copyright 1999 Harcourt Publishers Ltd. PMID- 10833565 TI - Thrombolysis in Acute Stroke Pooling Project: a meta-analysis on individual patient data. AB - To assess the efficacy and safety of thrombolytic therapy in acute ischaemic stroke, randomised clinical trials have been undertaken. Their results suggest that further research should be attempted to identify patients for whom the benefit/risk ratio of thrombolysis is beneficial. The Thrombolysis in Acute Stroke Pooling Project (TAS-PP) group will pool individual patient data from recent studies and meta-analyse these. A Steering Committee drafted the protocol and defined access rules to the common file. The objectives are to assess the efficacy of thrombolysis to reduce death or severe disability, to identify predictors of death and haemorrhagic transformation, and to identify subgroups with a better response to treatment, using logistic regression, survival curve comparison (log rank test), multivariate modelling (with treatment, baseline characteristics, delay from symptom to treatment as covariates). This project will help defining subpopulations that are more likely to benefit from this treatment, which cannot be achieved using tabulated data, and designing future trials. Copyright 1999 Harcourt Publishers Ltd. PMID- 10833566 TI - Value of homonymous and heteronymous monosynaptic reflexes in the diagnosis and follow-up of cervical spinal injuries. AB - Needle electromyographic examination constitutes the only neurophysiological test used routinely to assess the integrity of proximal nerve pathways. A heteronymous reflex, with a latency consistent with a monosynaptic pathway, can be evoked in the contracting biceps brachii muscle by stimulation of the median nerve at the elbow. This reflex response provides a complementary technique to the biceps and supinator jerks for accessing the C(5)/C(6)motoneuron pool because its afferent limb involves the median nerve and the C(5)/C(6)posterior roots. Three cases are described to illustrate the clinical value of this heteronymous monosynaptic reflex of biceps brachii and of conventional H reflexes in the diagnosis and follow-up of lesions involving the C(5)/C(6)segments. Reflex function can be assessed for most spinal segments innervating the upper and lower limbs, thus allowing both objective confirmation of the clinical findings and documentation of changes over time. Copyright 1999 Harcourt Publishers Ltd. PMID- 10833567 TI - Resection of large petroclival meningiomas by the simple retrosigmoid route. AB - The operative treatment of petroclival meningiomas has shown considerable improvement in recent years due at least in part to new developments in skull base techniques, including the introduction of the modern transpetrosal approaches. Increased tumour resection and improved postoperative outcomes have been reported with the use of the transpetrosal approaches. Extensive experience in managing these tumours has shown, however, that in a subgroup of patients the transpetrosal approaches may not be indicated: either because of the increased surgical complications related to this approach, or because of the unnecessarily large exposure in the individual cases. The authors report a recent experience dealing with 70 petroclival meningiomas, 24 of which have been resected by the simple retrosigmoid approach. The indication for the retrosigmoid route in the surgical management of the petroclival meningiomas, and its advantages and disadvantages compared to the transpetrosal method are discussed. Copyright 1999 Harcourt Publishers Ltd. PMID- 10833568 TI - Biomodelling of skull base tumours. AB - Stereolithographic (SL) biomodelling is a new technology that allows three dimensional computed tomography magnetic resuance data to be used to generate solid plastic replicas of anatomical structures (biomodels). A prospective trial of 11 patients to assess the utility of SL biomodelling in skull base tumour surgery has been performed. Biomodels were manufactured and used for patient education, diagnosis and operative planning. The results suggest that biomodelling has significant utility in skull base tumour surgery and is a cost effective technology. Biomodels were found to be particularly effective for surgical simulation and the production of customised cranioplastic implants for reconstruction after tumour resection. Copyright 1999 Harcourt Publishers Ltd. PMID- 10833569 TI - Surgical complications of functional neurosurgery treating movement disorders: results with anatomical localisation. AB - Thalamic and pallidal lesions can alleviate movement disorders, but to achieve this safely and efficaciously requires accurate target localization. We report the surgical complications encountered using an anatomical localization technique to create 121 thalamic and pallidal lesions in 79 consecutive patients over a 3 year period. There was no perioperative mortality, although there was one late death indirectly related to surgery. The risk of haemorrhage was 3.3% per lesion made. Anatomical localization offers a relatively safe way of identifying targets for functional neurosurgery, with complication rates which compare favourably with the published literature. Copyright 1999 Harcourt Publishers Ltd. PMID- 10833570 TI - Laboratory studies. PMID- 10833572 TI - Case reports. PMID- 10833571 TI - Balo's concentric sclerosis in a woman from Papua New Guinea. AB - We report a case of Balo's concentric sclerosis (a variant of multiple sclerosis) from Papua New Guinea. A 42-year-old woman with a past episode of optic neuritis presented with a left hemiparesis. Magnetic resonance imaging revealed a solitary large tumour-like right cerebral lesion with a pattern of concentric bands of different signal intensities. The diagnosis was established by biopsy of the lesion. To our knowledge, this is the first reported case of Balo's concentric sclerosis in the indigenous population of Papua New Guinea. Copyright 1999 Harcourt Publishers Ltd. PMID- 10833573 TI - A novel hybrid biological embolic material: autologous fibroblast incorporated collagen (FC) beads. AB - If viable fibroblasts are used as an active ingredient of an embolic material, it is expected that proliferation and extracellular matrix production of fibroblasts incorporated at a vascular lesion will help restore tissues due to endovascular scar formation, resulting in progressive and permanent occlusion. Based on this working principle, we have devised a novel biological embolic material, hybrid fibroblast incorporated collagen (hybrid FC) beads composed of collagen microbeads and autologous fibroblasts harvested from the subcutaneous tissue of the host to be treated. Hybrid FC beads were prepared by culture of fibroblasts harvested from canine subcutaneous tissue on collagen microbeads (diameters ranging from around 100 um to around 400 um). Canine kidneys were embolized with either hybrid FC or cell-free collagen beads via a transarterial route. Histological examination up to 6 months after embolization revealed that, although both embolic materials effectively occluded the target vessels at the time of embolization, intravascular scar formation activity at the embolized sites was much more profound in the case of the hybrid FC beads than in that of the cell-free beads. Proliferation of autologous fibroblasts was verified by the expression of alkaline phosphatase activity of gene-transfected fibroblasts at the site of lodgement of the beads. It is expected that, using the novel hybrid biological embolic material, hybrid FC beads used for vascular lesions such as arteriovenous malformations can be treated more effectively to restore tissues, resulting in minimal recanalization which often occurs when synthetic embolic materials are used. Copyright 1999 Harcourt Publishers Ltd. PMID- 10833574 TI - The management of acute neurotrauma in rural and remote locations: A set of guidelines for the care of head and spinal injuries. AB - The Neurosurgical Society of Australasia through its Trauma Committee has a long involvement in the problem of neurotrauma. The management of acute neurotrauma in rural and remote locations is of particular interest and is part of a general policy which includes education, prevention, organization of an integrated neurotrauma system and support for the Early Management of Severe Trauma (EMST) programme instituted by the Royal Australasian College of Surgeons. The management of acute neurotrauma requires a consultative approach especially in the multiple injured patient and where transfer or retrieval is necessary. As acute neurotrauma may present to general practitioners, rural surgeons or emergency departments in country hospitals, a set of guidelines has been developed to assist in the early management of acute neurotrauma. These guidelines have been previously published by the Neurosurgical Society of Australasia and the Royal Australasian College of Surgeons. They have been extensively used since 1992 by rural health and distance education groups, Royal Flying Doctor Service Emergency Management of Severe Trauma Course of the Royal Australasian College of Surgeons and by overseas education programs for neurotrauma care. This revision updates the guidelines and makes them accessible through the Journal. Reprints are available for practitioners in rural and remote settings, trauma course participants and others. Copyright 1999 Harcourt Publishers Ltd. PMID- 10833575 TI - The origin of the concept of partial epilepsy. AB - The International League Against Epilepsy has devised classifications which subdivide both epileptic seizures and the epilepsies and epileptic syndromes into two main types: generalized and partial. Epileptogenesis in the partial variety is believed to originate in a localized part of the cerebral cortex and results in clinical manifestations which appear to commence in only a restricted part of the sufferer's body. Use of the term 'partial' in relation to these entities has often been said to date back to James Cowles Prichard (1786-1849) who was the author of the second major work on epilepsy to be written in the UK. While Prichard certainly described 'partial epilepsy', he stated that he intended the words to refer to the fact that the disorder he described under that designation was only partly, and not fully, epileptic in nature. He did not refer to the fact that it affected only part of the body as his basis for using the term. In the absence of knowledge of localization of function in the cerebral cortex at Prichard's time of writing, he had no basis for deducing that the underlying epileptic process arose in only part of the brain. However, there is an earlier mention of the use of the word 'partial' in relation to epilepsy. This is to be found in the writings of the great Scottish physician William Cullen (1710-1790), and there is reason to believe that Prichard should have been aware of this. Cullen used 'partial' with an intention similar to the modern one, employing the word to refer to seizures which affected only part of the body. Credit for the origin of the idea of a 'partial' epilepsy should belong to Cullen; not only did he have priority over Prichard but his concept was closer to the modern one than was Prichard's. Copyright 1999 Harcourt Publishers Ltd. PMID- 10833576 TI - Contemporary psychosurgery. AB - Psychosurgery is a safe and relatively effective treatment which should be offered to patients with intractable obsessive compulsive disorder (OCD), major affective disorders, and chronic anxiety states after a minimal period of 2 to 5 years and after all other reasonable treatments have been tried. A greater understanding of the biological basis of psychosurgery is developing from advances in functional brain imaging. The optimal site and size of the lesions remains to be established. The standard technique uses stereotactic radiofrequency thermolesions but stereotactic radiosurgery is an alternative. A good outcome following psychosurgery can be expected in 50-60% of cases, and some of the failures may respond to reoperation. This leaves about 40% who have not benefited or in a few cases may be worse. Unfortunately, there are as yet no reliable clinical or investigational predictive indicators for the good outcome group. Restricted accurate lesion placement minimizes personality change, epilepsy and cognitive decline. Careful safeguards, including approval by an independent multidisciplinary legally constituted review board are mandatory in selecting and following the patients, but unfortunately, there are individuals who are never referred or reside in a society which proscribes the operation, whose lives could be made more tolerable. Psychosurgery is probably underutilized due to negative perceptions based on historical factors, and strict regulations and legislation limiting its application. It is possible that with advances in psychiatric pharmacotherapy, the psychosurgery procedures will be made redundant, but we believe that until this eventuates there is still a small place for these operations. Psychosurgery should only be carried out in a national centre by a multidisciplinary team with experience in these disorders, and an intensive ongoing postoperative rehabilitation program is essential to achieve the best results. Copyright 1999 Harcourt Publishers Ltd. PMID- 10833577 TI - Asymmetries of cerebral perfusion in a stroke-age population. AB - Cerebral perfusion measurements with(99m)Tc-hexamethylpropyleneamine oxime single photon emission computed tomography (HMPAO SPECT) are potentially clinically useful in the investigation of patients with acute ischaemic stroke. The normal side-to-side asymmetry on(99m)Tc-HMPAO SPECT images may be greater in the stroke age population than in younger age groups. To assess the extent of variation we studied 66 volunteers with(99m)Tc-HMPAO SPECT who were closely age matched to a stroke population and who had normal CT and extracranial and intracranial vascular ultrasonography. By measuring the side-to-side ratios in 19 regions of interest, the normal side-to-side perfusion asymmetry was determined and, as a secondary aim, variations in regional asymmetry were correlated with age. After repeated measurement analysis of variance (ANOVA), the normal range of side-to side ratios in this population was 1.003+/-0.145 (mean+/-1.96 SD, ANOVA). The two regions with the greatest side-to-side asymmetry values were (1) the superior lateral temporal cortex (higher on the right relative to the left) and (2) the superior parietal cortex (higher on the left relative to the right). The orbitofrontal and posterior temporal cortices demonstrated significantly increased bidirectional variation with age (P< 0.05). In a stroke-age population hemispheric perfusion is relatively symmetrical and a side-to-side variation of greater than +/-14.5% may be defined as pathological. This value is higher than that reported in younger volunteers suggesting that there is a mild increase in the side-to-side perfusion asymmetry with age. Increased variation in the orbitofrontal and posterior temporal cortices occurs with age and should be borne in mind when interpreting HMPAO SPECT studies. Copyright 1999 Harcourt Publishers Ltd. PMID- 10833578 TI - Semiautomated analysis of the extent and severity of perfusion defects on brain SPECT images: validation studies. AB - Single photon emission computed tomography (SPECT) is a widely available and practical functional imaging technique with established clinical and research applications in neurological disorders such as epilepsy and stroke. SPECT images are usually analysed visually, or semiquantitatively by measuring side-to-side asymmetries. In order to evaluate perfusion change after thrombolytic therapy in patients with acute ischaemic stroke we developed a semiautomated, weighted volumetric analysis (the ischaemic index) that measured the extent and severity of tracer uptake abnormalities on brain SPECT images semiquantitatively. The unaffected cerebral hemisphere was used as the reference region of interest. The analysis was validated in a phantom brain model incorporating 'strokes' varying in size and degree. The phantom 'stroke' sizes measured with the ischaemic index analysis correlated closely with the true values (r=0.994, P< 0.01) and this correlation was maintained under low count conditions acquired to simulate the clinical setting. The overall operator dependent error of the analysis was +/ 3.4%. In 30 patients treated with thrombolytic therapy who were studied serially with(99 m)Tc-hexamethylpropyleneamine oxime (HMPAO) SPECT, the analysis was used to measure hypoperfusion volume and provide indices of perfusion change. This analysis has the advantages of semiautomation, ease of use and validation and has a potentially wide range of applications for both SPECT and PET. Copyright 1999 Harcourt Publishers Ltd. PMID- 10833579 TI - Diagnosis of spinal cord ependymoma and astrocytic tumours with magnetic resonance imaging. AB - Ependymoma and astrocytic tumours, both originating from the glial tissue, constitute the majority of intramedullary spinal cord tumours. The purpose of this study is to evaluate the diagnostic ability of magnetic resonance imaging (MRI) in these two types of intramedullary spinal cord tumours. MRI findings of 17 ependymomas and 11 astrocytic tumours of the spinal cord were reviewed. Pre contrast T(1)-weighted images revealed abnormal enlargement of the spinal cord in all cases. Peritumoural cyst formation was noted in 14 ependymomas (82%) and four astrocytic tumours (36%). Evidence of haemorrhage was identified in five (29%) ependymomas and one (9%) astrocytic tumour. Gadolinium-MRI (Gd-MRI) revealed marked enhancement in 11 ependymomas (65%), while six ependymomas showed irregular, partial or no enhancement. Five of these six ependymomas had evidence of prior haemorrhage. Eight astrocytic tumours (73%) were enhanced on Gd-MRI and the pattern of enhancement was more irregular or partial than that of ependymomas. Two pilocytic astrocytomas showed marked ring-like enhancement. In conclusion, detection of haemorrhage and Gd-MRI will help to differentiate these two types of tumours. Further study will be needed to elucidate the relationship between the histological subtypes of the tumours and MRI findings. Copyright 1999 Harcourt Publishers Ltd. PMID- 10833580 TI - Paediatric intracranial aneurysms: results of a surgical series and literature review of Guglielmi detachable coil embolization. AB - It is known that the profile of aneursyms in the paediatric age group demonstrates a different biology and epidemiology than those in the adult population in terms of size, sex, location, multiplicity and complications such as vasospasm and rebleeding. As a consequence of their different behaviour, timing of surgery does not appear to be such a critical issue in their management decision. Surgical clipping is still the treatment of choice, however, with our growing experience of endovascular techniques, these might prove to be a suitable alternative in the future. We report the results of our surgical series of 17 paediatric and adolescent patients harbouring 18 arterial intracranial aneurysms (age range 7-18 years). Systemic diseases traditionally known to be associated with this entity were excluded. In order to assess endovascular techniques as a potential treatment alternative for this group of patients, we reviewed the current literature, dealing with Guglielmi detachable coil (GDC) embolization of arterial cerebral aneurysms in this age group. No current series deals purely with paediatric aneurysms; we have reviewed paediatric cases reported as part of large adult series. Copyright 1999 Harcourt Publishers Ltd. PMID- 10833581 TI - Carotid endarterectomy in a neurosurgical unit: a retrospective review. AB - A retrospective survey of 262 carotid endarterectomies performed in the Neurosurgical Unit at Royal Prince Alfred Hospital over a 15 year period was conducted. A total of 141 patients were operated on for transient ischaemic attacks referable to stenosis of the ipsilateral internal carotid artery. Eighty eight percent of patients had angiographic evidence of greater than 80% stenosis of the affected internal carotid artery. A postoperative new permanent neurological deficit of 1.9% and death rate of 0.6% was achieved. No deaths and two postoperative strokes were recorded after 1987. These results are comparable with published series. The use of barbiturate therapy with electroencephalography changes on cross clamping of the internal carotid artery is described. Copyright 1999 Harcourt Publishers Ltd. PMID- 10833582 TI - Perivascular CSF flow in the rat cerebellum. AB - Cerebrospinal fluid (CSF) flow from the subarachnoid space into the cerebellum was studied in rats using Horseradish peroxidase (HRP) injected into the cisterna magna. Animals were sacrificed after 0, 10 or 30 min Transpial diffusion of HRP did not extend beyond the Purkinje cell layer. At all time intervals, perivascular spaces labelled with HRP were observed in cerebellar grey and white matter. In the cerebellar granular layer and white matter, HRP was usually present only in perivascular spaces and the surrounding extracellular space. There was no evidence of a preferential flow towards the fourth ventricle. These results suggest that there is a perivascular pathway for rapid flow of CSF from the subarachnoid space into the perivascular spaces and then into the cerebellar extracellular space. The Purkinje cell layer appears to act as a barrier to diffusion from the pial surface. Copyright 1999 Harcourt Publishers Ltd. PMID- 10833583 TI - Ventriculocisternal drainage via endoscopic third ventriculostomy after endovascular embolization for aneurysmal subarachnoid haemorrhage. AB - Coil embolization is a favoured method of treating acutely ruptured aneurysms. However, subarachnoid clots cannot be removed with endovascular treatment, which is disadvantageous from the view point of preventing vasospasm. The authors report intrathecal fibrinolytic therapy via a ventricular drainage tube instituted after endoscopic third ventriculostomy for effective prevention of vasospasm after coil embolization. Two cases of poor grade aneurysmal subarachnoid haemorrhage were treated with interlocking detachable coils. Following the acute stage embolization, endoscopic third ventriculostomy was performed and a drainage tube was inserted into a lateral ventricle or basal cistern. Intrathecal fibrinolytic therapy with Urokinase was performed via the drain tube to a spinal drain. In both cases, the subarachnoid clot disappeared rapidly without any clinical signs of vasospasm. Copyright 1999 Harcourt Publishers Ltd. PMID- 10833584 TI - Neuropathology of three clinical cases prospectively diagnosed as dementia with Lewy bodies. AB - Concensus guidelines for the clinical and pathological diagnosis of dementia with Lewy bodies have recently been proposed based on retrospective studies (McKeith IG, Galasko D, Kosaka K et al. Consensus guidelines for the clinical and pathologic diagnosis of dementia with Lewy bodies (DLB): report of the consortium on DLB international workshop. Neurology 1996; 47: 113-1124.) The present study tests these criteria prospectively in three cases which came to autopsy: a 57 year-old female followed for 5 years, an 86-year-old male followed for 3 years and a 66-year-old male followed for 11 years. All were considered to have dementia with Lewy bodies clinically, and at autopsy all had pathologically confirmed Lewy body disease. However, the Lewy bodies found in the 57-year-old were scanty and she also had marked Alzheimer-type pathology, making the contribution of Lewy bodies to the dementia uncertain. The 66-year-old had unusual cortical Lewy body pathology, previously only described in one other case (Masliah E, Galasho D, Wiley CA, Hansen LA. Lobar atrophy with dense-core (brain stem type) Lewy bodies in a patient with dementia. Acta Neuropathol 1990; 80: 453 458.) While this study prospectively validates the current criteria for dementia with Lewy bodies, there was considerable pathological variability in the cases. Copyright 1999 Harcourt Publishers Ltd. PMID- 10833585 TI - Case reports. PMID- 10833613 TI - Academic neurology in Australia. PMID- 10833614 TI - Anti-GM1 ganglioside antibodies: their role in the diagnosis and pathogenesis of immune-mediated motor neuropathies. PMID- 10833615 TI - Recent advances in the pre-mortem diagnosis of Creutzfeldt-Jakob disease. AB - Included in the spectrum of human transmissible spongiform encephalopathies are Creutzfeldt-Jakob disease (CJD) and the new variant form (vCJD), Gerstmann Straussler-Scheinker syndrome, fatal familial insomnia, kuru and various less distinct neuropsychiatric disorders. Progress in our understanding of this group of disorders continues at a prodigious rate, although important vexing practical issues persist. The definitive confirmation of symptomatic prion disease still requires pathological examination, most reliably performed post-mortem. However, paralleling the recent advances in the molecular biological understanding of normal prion protein (PrP(c)) function and the pathophysiology of prion diseases, there have been worthwhile developments in the pre-mortem diagnosis of CJD. Efforts to develop less invasive but very reliable ante-mortem diagnostic tests have received an additional impetus because of the potential epidemic of vCJD. Historically, the ancillary investigation of most merit has been the EEG, whereas the recent advances have encompassed a broader range of technologies, including both magnetic resonance and radioisotopic neuroimaging, and immunoassays for a range of non-specific marker proteins in both CSF, and less commonly, blood. However, given the recent refinement of sophisticated immunoassays, it is envisaged that the pathognomonic, protease-resistant, disease-associated isoforms of the prion protein (PrPres) may soon be directly detectable in the blood and tissues of patients manifesting or incubating a spongiform encephalopathy. PMID- 10833616 TI - Creutzfeldt-Jakob disease: diagnostic utility of 14-3-3 protein immunodetection in cerebrospinal fluid. AB - With the aim of improving the pre-mortem diagnostic accuracy of sporadic Creutzfeldt-Jakob disease (CJD), there has been considerable recent interest in the merit of immunodetecting 14-3-3 proteins in the cerebrospinal fluid (CSF) using Western blotting, with cumulative support for the utility of this technique. As a corollary, during a 20 month period, CSF samples from an unselected prospective series of 124 patients in whom sporadic CJD was a differential diagnostic possibility were examined by the Australian Creutzfeldt Jakob disease Registry (ACJDR) for the presence of 14-3-3 proteins. Follow up to achieve a final diagnosis or clinical outcome was successful in 119. For definite and probable sporadic CJD combined, a positive result was 91.4% sensitive, while the sensitivity for the pathologically verified group alone was 96.0%. A negative outcome was 92.5% specific with false positive results seen in five patients with diagnoses which included inflammatory CNS disorders, cerebral ischaemia and dementia with Lewy bodies (DLB). Immunodetectable 14-3-3 proteins were present in three of four symptomatic patients with prion protein gene (PRNP) mutations. CSF samples containing significant amounts of blood were confirmed as suboptimal, with weak or qualitatively unusual positive results found in greater than 50% of such specimens, with only one of 14 such cases ultimately classified as definite or probable CJD. PMID- 10833617 TI - Results of testing for anti-GM1 antibodies. AB - We used an ELISA technique to measure IgG and IgM antibodies to the ganglioside GM1, with the results expressed in arbitrary units. We tested 1007 sera from patients with peripheral neuropathy or muscle weakness. For IgG and IgM antibodies, the distribution of results differed significantly from a normal distribution. In the patient group, 81 of 1007 sera had elevated levels of IgG antibodies (> 10 units). Of these, 11 patients had very high levels (> 50 units). These 11 patients had diagnoses of GBS (4), motor neurone disease (3) or non specific idiopathic neuropathy (4). For IgM antibodies, 115 of 1007 sera were positive (> 20 units). Of these, 18 patients had very high levels (> 50 units). These 18 patients had diagnoses of Guillain-Barre syndrome or Miller Fisher syndrome (4), multifocal motor neuropathy (4), motor neurone disease (2), non specific neuropathy (2). We conclude that anti-GM1 antibodies in high titre are uncommon. Patients with multifocal motor neuropathy have high levels of antibody. However, patients with other disorders may also have high levels, so that anti GM1 antibody levels alone are not a specific test for multifocal motor neuropathy. We found that antibodies to GM1 were present in the sera of patients with chronic idiopathic neuropathy, leading us to suggest that these antibodies may sometimes arise as a secondary response to disease. PMID- 10833618 TI - Significant anticonvulsant side-effects in children and adolescents. AB - The anticonvulsant (AED) history for 216 children and adolescents with epilepsy was reviewed to determine the incidence and types of significant side effects (SSE) which warranted ceasing the drug (not due to a lack of response or a high dose). All parents of patients with epilepsy seen by the author over a 2 year period (March 1996 - March 1998) were questioned about SSE to previous AEDs, and the child's current therapy was also monitored prospectively to determine SSE. There were 107 girls and 109 boys ranging in age from 3 months - 18 years. Eighty three patients had been exposed to a single AED while 133 had multiple AED exposures: mean 3.6 drugs; range 2-10 drugs. They were exposed to a total of 568 AEDs with SSE occurring in 15% of drug contacts: 7% due to behavioural changes such as irritability, aggression or hyperactivity; 8% were due to other factors such as a rash, headache, gastrointestinal disturbance or drowsiness. Fifty-seven children (26%) had experienced at least one SSE with 19 (9%) having SSE to more than one AED (range 2-4). Global developmental delay or an intellectual disability (ID) were present in 67 patients, and 27 (40%) of these experienced SSE compared with 30 (20%) of the group with normal cognition. This difference was principally due to the higher incidence of behavioural SSE in the ID group 28% versus 6% for the normal cognition group. Allowing for the higher number of AEDs used in the ID group (implying that their epilepsy was more difficult to control), behavioural SSE were still significantly more likely to occur in this group, i.e. 1: 9.6 drug exposures compared with 1: 31. 8 exposures for the normal cognition group (P<0.001). Monotherapy trials underestimate the true incidence of SSE in clinical practice as 26% of children had experienced at least one SSE and 9% had SSE to more than one AED. Those with ID were three times more likely to have behavioural SSE than children with normal cognition. PMID- 10833619 TI - Extensive cervical laminoplasty for patients with long segment OPLL in the cervical spine: an alternative to the anterior approach. AB - We investigated treatment of long segment cervical OPLL by posterior decompression using a laminoplasty technique. Our aim was to both decompress the spinal cord and also to preserve neck motion. There were 38 patients treated by this posterior approach. Twenty-eight patients underwent C1-C7 expanding laminoplasty, 4 patients underwent C1-T1 expanding laminoplasty, and 6 patients C2-C7 expanding laminoplasty. The transverse width of the open-door laminoplasty was sufficient to achieve decompression of not only the spinal cord but also the nerve root outlets at each laminoplasty level. There were no complications related to this surgical technique, nor late deterioration in the mean follow up period of 4. 5 years. We propose expanding laminoplasty as an important option for the treatment of long segment cervical OPLL. PMID- 10833620 TI - The changing pattern of head injury in Thailand. AB - OBJECTIVE: To determine whether patterns of head injury are changing with time. MATERIALS AND METHODS: A total of 3194 and 4217 consecutive trauma patients who attended the emergency room in 1985-86 and 1996 respectively were studied with respect to age, sex, cause of injury, injury severity, pathology, and outcome. RESULTS: The number of patients with head injury in 1996 nearly doubled (1224/4,217:29.03%) when compared to the 1985-86 study (504/3, 194; 15.78%). This was due to an increase in the outpatient subgroup (1009/1224). The admitted patients with head injury showed a pattern of less severe injury. Severe head injury decreased from 12.4 to 7. 9%. However, acute subdural haematoma and diffuse brain injury increased from 12.2% and 9% to 32% and 16.8% respectively. The mortality rate of admitted patients increased statistically significantly from 14.4% to 21.8% between the 1985-86 and 1996 studies. CONCLUSIONS: This comparative study showed attend toward less severe injury. This may be due to multiple factors. The predominant factor may be the compulsory use of motorcycle helmets. The limitation of this study was that it utilised tertiary hospital based data only. Tertiary hospital receive more and serious head injured patients from surrounding provincial hospitals this may be the major cause of the increased the mortality rate. PMID- 10833621 TI - Cerebral oxygenation and systemic trauma related factors determining neurological outcome after brain injury. AB - We examined the relationship between clinical and radiological findings, cerebral oxygenation patterns during intensive care management, presence of systemic trauma related injuries and severity of illness in 50 patients (age: 32.3 +/- 12 years, GCS: 8 +/- 4) who were rescued from the accident scene within a 30 min period after trauma. Presence of systemic injuries was quantified using the Injury Severity Score (ISS) and severity of illness was scored using the Acute Physiology and Chronic Health Evaluation (APACHE II). Cerebral oxygenation parameters included continuous monitoring of jugular bulb oxygen saturation (SjvO(2)) for 12 840 h, and 2323 periodical blood sampling for measurement of arteriovenous differences in oxygen content (AVDO(2)), arteriovenous difference of lactate (AVDL) and lactate oxygen index (LOI). Fifteen patients (30%) presented with anisocoria or non-reacting pupils. Diffuse lesions on computed tomography (CT) were found in 34% of the patients and in 66% a mass lesion was removed. The mean ISS was 28 +/- 15.3 and 34 patients (68%) had an APACHE II score between 20 and 29 (mean 24 +/- 15). No statistically significant association between age (P = 0.45), gender (P = 0.83), initial Glasgow Coma Score (GCS) (P = 0.43), episodes of cerebral perfusion pressure (CPP) < 70 mm Hg (P = 0.8), ISS (P = 0.28), pupillary abnormalities (P = 0.57), initial CT findings (P = 0.74), APACHE II scores (P = 0. 36) and outcome could be demonstrated. The number of SjvO(2)desaturations (< 60%) was the only statistically significant factor associated with outcome (P = 0.05). The percentage of patients with poor neurological outcomes (GOS 1-3) was 38% in patients with no or one desaturation episode, and 57.6% in those with multiple desaturations. In conclusion, in patients who are resuscitated early and quickly transferred to the hospital, the number of SjvO(2)desaturations during intensive care management might be associated with outcome more strongly than other clinical and radiological features. PMID- 10833622 TI - Intraoperative monitoring to preserve central visual fields during occipital corticectomy for epilepsy. AB - Photic driving using a flashing strobe light was recorded via intracranial electrodes in two patients with occipital epilepsy being evaluated for surgery. The same technique was used to monitor the visual cortex intraoperatively. Visual evoked potentials (VEPs) were also obtained using the intracranial electrodes in one patient. Preoperative photic driving occurred in a separate location from the cortical areas producing ictal epileptiform activity. VEPs were located in the same site as photic driving. Photic driving was monitored throughout the resection and remained unaffected at the end of each procedure. Postoperative visual field testing in both patients showed preservation of central vision although some reduction in peripheral fields was seen. Intraoperative monitoring of the visual cortex using photic stimulation proved to be a reliable technique for preserving central vision during occipital lobe surgery. PMID- 10833623 TI - The early protective effects of L-arginine and Ng-nitro-L-arginine methyl ester after experimental acute spinal cord injury. A light and electron microscopic study. AB - The purpose of this study was to investigate the early protective effects of L arginine and Ng-nitro-L-arginine methyl ester (L-NAME) after acute spinal cord injury. Acute spinal cord injury was performed by epidural application of an aneurysm clip at thoracic (T) 7 - 11 level. L-arginine at a dose of 750 microg/kg/min was administered 10 min before acute spinal cord injury and continued for 30 min to 10 animals (Group II). L-NAME at a dose of 250 microg/kg/min was administered 10 min before acute spinal cord injury and continued for 30 min to 10 animals (Group III). No drug was administered to 10 animals after acute spinal cord injury (Group I). Light and electron microscopic analysis were performed in all of the groups. Oedema of perineural, axoplasm or white matter in the L-arginine-treated group was less than in Group I and Group III. Thickening in the walls of the arterioles and venules in the L-arginine treated group was much milder than in Group I and Group III. Degeneration of myelinated axons in the L-arginine-treated group was milder than in the control group. But there was no different between Group II and Group III. PMID- 10833624 TI - Endovascular treatment of intracranial aneurysms with Guglielmi Detachable Coils: emphasis on new techniques. AB - Endovascular therapy for intracranial aneurysms has evolved since Serbinenko pioneered embolisation with latex balloons in the 1970s. The focus of modern endovascular therapy has shifted to the use of Guglielmi Detachable Coils (GDC; Boston Scientific Corporation, Natick, MA, USA) which are retrievable until the operator is satisfied with placement and they are detached. GDC therapy has been shown to be most efficacious in smaller aneurysms with relatively large dome:neck ratios which allow maximal coil packing within the aneurysm lumen. Wide neck aneurysms with dome:neck ratios of less than 2.0 and large aneurysms have a significantly lower incidence of complete treatment, with higher rates of repeat rupture following GDC therapy. The geometry of wide neck aneurysms is less favourable for retention of coils within the aneurysm lumen, resulting in greater risk of parent vessel compromise from coil herniation and difficulty obtaining maximal coil packing. This chapter will summarise GDC therapy for intracranial aneurysms including newer techniques designed to address the problem of wide neck aneurysms. PMID- 10833625 TI - Cerebral arteriovenous malformations associated with aneurysms--a report of 10 cases and literature review. AB - We analysed 10 cases of arteriovenous malformations (AVMs) associated with aneurysms and reviewed the literature in order to confirm the clinical characteristics of AVMs associated with aneurysms. The mean incidence of aneurysms in our AVM population was 9.1% and the average age was 38 years. The ratio of male to female was 1.3 to 1. The aneurysms (40-96%) were often located on a feeding artery and seldom (23%) at an arterial bifurcation. The mortality rate was 0-26% in surgically managed patients and 38-60% in conservatively treated patients. The combination of intracranial vascular lesions is not rare. Most patients presented with symptoms referable to their AVMs. Treating both lesions in a single operation is the best option. The prognoses for most patients was good. PMID- 10833626 TI - Parkinsonism after correction of hyponatremia with radiological central pontine myelinolysis and changes in the basal ganglia. AB - Parkinsonism has been rarely described following central pontine and extrapontine myelinolysis. We report a case of parkinsonism developing following rapid correction of hyponatremia with radiological evidence of central pontine myelinolysis and changes in the basal ganglia. A 56-year-old man developed drooling and bilateral hand tremors 3 weeks after correction of hyponatremia from 103 to 125 mmol/L over 14 h. He had a prominent 6 Hz resting tremor which worsened with action and mild cogwheel rigidity. Magnetic resonance imaging (MRI) showed changes consistent with central pontine myelinolysis and increased signal on T1-weighted images in the putamen bilaterally. His tremor responded well to L dopa therapy. There have been several other cases of parkinsonism developing after central pontine/extrapontine myelinolysis. Increased signal in the basal ganglia on T1-weighted images has been described in another case of central pontine myelinolysis imaged about the same time after sodium correction as our case. PMID- 10833627 TI - Emergency surgical recanalisation of A1 segment occluded by a Guglielmi detachable coil. AB - We describe a case of A1 segment occlusion caused by a tangled Guglielmi detachable coil (GDC) during an endovascular treatment for a ruptured anterior communicating aneurysm. Immediate surgical recanalisation of the occluded artery and clipping of the aneurysm rescued the patient. This case demonstrates that an emergency surgical team, as well as an operating room, should be available during any complicated intra-aneurysmal procedure, so that timely surgical intervention could be carried out without delay. PMID- 10833628 TI - Head trauma and brain tumours revisited. AB - The authors report a case of an anaplastic astrocytoma which on magnetic resonance imaging and direct visualisation was continuous with an area of gliosis in the left frontal lobe. This gliosis was secondary to a head injury received 19 years earlier that required evacuation of an intracerebral haematoma. This case largely meets the accepted criteria for brain tumour associated with head trauma. PMID- 10833629 TI - Late intramedullary spinal cord metastasis in a patient with lymphoblastic lymphoma: case report. AB - Late intramedullary spinal cord metastasis of lymphoblastic lymphoma (LBL) is unreported in the literature. The central nervous system (CNS) sites of metastasis previously documented include the spinal leptomeninges causing epidural spinal cord compression or brain praenchymal sites within a year of primary diagnosis. This report represents the first case of intramedullary spinal cord LBL as a late recurrence. The method consists of a case study of one patient with retrospective analysis of tumour tissues from biopsies at ages 5, 9 and 26. The results show that late spinal cord recurrence was microneurosurgically subtotally debulked and later treated with radiotherapy with improvement in neurological deficit. Investigations revealed no evidence of systemic disease and this recurrence appears to be an isolated sanctuary site tumour. A MEDLINE search (1966 - present) of the literature failed to reveal similar reported cases and the authors believe this to be the first late intramedullary spinal cord metastasis in a patient with LBL. PMID- 10833631 TI - Diary of events PMID- 10833630 TI - Fascicular arrangement within the oculomotor nerve MRI analysis of a midbrain infarct. AB - The fascicular arrangement of the oculomotor nerve within the midbrain is not adequately elucidated in humans. We treated a patient with a partial oculomotor palsy who had impaired adduction and supraduction on the left side, which were attributed to an ipsilateral lacunar infarct. CT and MRI revealed a discrete lesion in the centre of the midbrain tegmentum in the rostrocaudal plane. This case suggests that the oculomotor fibres for extraocular movement are located in the middle of the the midbrain, and supports the fascicular proximity of the superior and medial rectus muscles. The fascicular arrangement of the midbrain oculomotor nerve is speculated to be pupillary component, extraocular movement and eyelid elevation in that rostrocaudal order, based on the previous reports of neuro-ophthalmological impairment and MRI findings, which are analogous to the nuclear arrangement proposed by Warwick. PMID- 10833632 TI - News and notices PMID- 10833633 TI - The effect of carnosine on rats during experimental brain ischemia. PMID- 10833634 TI - Membrane and soluble forms of intestinal disaccharidases in young rats whose mothers received low-protein food during pregnancy and lactation. PMID- 10833635 TI - Interval training by normobaric hypoxia accelerates the reinnervation of musculus extensor digitorum longus in mice. PMID- 10833636 TI - Interdependence between the parameters of human immunity and higher nervous activity. PMID- 10833637 TI - Notch-noise testing of auditory neuron receptive fields. PMID- 10833638 TI - Organization of the retinal ganglion layer and retinal resolution in the sea otter (Enhydra lutris). PMID- 10833639 TI - Connection between the changes of derivatives of albumin spectra and the accuracy of determination of the albumin-globulin coefficient in the case of radiation damage. PMID- 10833640 TI - The informational role of tremor in color vision. PMID- 10833641 TI - Sensory neurons of the cat stellate ganglion. PMID- 10833642 TI - Epiphyseal peptides influence antibody formation in Peyer's glands of immunized rats. PMID- 10833643 TI - The intensity of emotional experience, the strength of the need, and the probability of its satisfaction: ECG and EEG parameters. PMID- 10833644 TI - A comparative ecological study of helminthofauna in sea waterfowl from Novaya Zemlya and eastern Murman. PMID- 10833645 TI - Prognosis of the spaciotemporal dynamics of disturbed ecosystems using three dimensional universal kriging based on sequential aerial and space surveys. PMID- 10833646 TI - How differences in basal metabolism affect energy expenditure on self-maintenance and energy efficiency in passeriformes and non-passeriformes. PMID- 10833647 TI - Temperature dependence of basic metabolism in plants with different heat requirements. PMID- 10833648 TI - Clonal populations of the hydroid Obelia longissima in the Arctic. PMID- 10833649 TI - On the targets of damaging environmental effects in a green leaf: the aftereffect of gamma radiation on chloroplast energetics. PMID- 10833650 TI - Recombinational properties of the chimeric recA genes (Pseudomonas aeruginosa/Escherichia coli): gene regions responsible for the recombinational activity of the protein encoded. PMID- 10833651 TI - What determines the mass-exponent factor of 3/4 in the allometric equations of basal metabolism in homoiothermal animals? PMID- 10833652 TI - Acoustic activity displayed in the agonistic behavior of great and light gerbils. PMID- 10833653 TI - The microalgal food spectrum of Daphnia longispina during the algal bloom of an eutrophic water body. PMID- 10833654 TI - Variations in the abiotic environment as a prerequisite for optimal Rana temporaria L. larval development. PMID- 10833655 TI - Seasonal changes in production of tumor necrosis factor by macrophages of hibernating ground squirrels. PMID- 10833656 TI - Mechanisms of adaptation to technogenic stress in plant cenopopulations (Taraxacum officinale s. l.). PMID- 10833657 TI - Development of the vestibular apparatus in mammals with different ecological characteristics. PMID- 10833658 TI - Genomic heterozygosity and human longevity. PMID- 10833659 TI - Larval adaptations in limbs of the Onychodactylus fisheri (Hynobiidae, Caudata). PMID- 10833660 TI - Criteria of ecological hazards due to anthropogenic effects on the biota: searching for a system. PMID- 10833661 TI - A mathematical model of origin multi-aberrant cell during spontaneous mutagenesis. PMID- 10833662 TI - Possible contribution of the ligand-receptor mechanism to the function of ciliated mechanosensory cells of mollusks. PMID- 10833663 TI - Reproduction by "budding" of the trophoblast cells in the mink implanting blastocysts. PMID- 10833664 TI - Effect of sphingosine on Ca(2+)-induced reactions of the HL-60 cells. PMID- 10833665 TI - Radiation-adaptive response in Escherichia coli: the polA, recA, and lexA dependence of induction and repair of single-strand DNA breaks after gamma irradiation. PMID- 10833666 TI - Protein p70 is an effector molecule for cytokine Tag 7. PMID- 10833668 TI - Estrogens and male reproduction. AB - Aromatase is the terminal enzyme responsible for estrogen biosynthesis; it is present in the endoplasmic reticulum membrane of steroidogenic cells in vertebrates. The aromatase gene is unique and its expression is regulated in a tissue- and more precisely, in a cell-specific manner via the alternative use of various promoters located in the first exons. The aromatase gene expression, and its transduction in a fully active protein not only in somatic cells but also in germ cells of rodent testes on one hand, and the widespread distribution of estrogen receptors (ER alpha and ER beta) in the genital tract of the male on the other hand, are clearly in favour of a physiological role for estrogens in the regulation of mammalian testicular functions. Moreover, the aromatase deficiency is associated for instance with severe bone maturation problems and sterility in mouse and man; but conversely, it is well known that estrogens in excess are responsible for the impairment of spermatogenesis. Therefore these female hormones (or the androgens/estrogens ratio) play a physiological role in the development and maintenance of male gonadal functions and seem to control especially the spermatid production (both qualitative and quantitative aspects) and epididymal sperm maturation. PMID- 10833667 TI - Participation of myosin in root pumping activity. PMID- 10833669 TI - Expression, regulation and function of AC133, a putative cell surface marker of primitive human haematopoietic cells. AB - To explore the physiological significance of AC133 expression on human haematopoietic cells, we phenotyped normal and malignant human haematopoietic cells for AC133 expression, evaluated the utility of AC133 for isolating human stem/progenitor cells in comparison to other known early haematopoietic cell markers, investigated the role of AC133 in regulating hematopoiesis, and evaluated the possibility that MYB might regulate AC133. We found that while human CD34+ progenitor cells expressed AC133, expression was rapidly downregulated during differentiation. In apparent contrast, AC133 mRNA was detectable in cells isolated from CFU-Mix, BFU-E, CFU-GM and CFU-Meg colonies. Human cord blood CD34+ cells expressed AC133 at higher levels than their normal bone marrow counterparts. In apparent contrast to normal primitive haematopoietic cells, the AC133 protein was undetectable on cells from 24 different human haematopoietic cells lines, even though the majority of these cells expressed AC133 mRNA. Since CD34, AC133 and the c-kit (KIT) receptor are all co-expressed on human stem/progenitor cells, we compared the ability of monoclonal antibodies directed against each of these proteins to isolate early progenitor cells. Using these antibodies and magnetized particles in a standard immunoaffinity isolation protocol, we found that anti-CD34 and anti-KIT MoAbs could isolate > 80-90% of the clonogeneic cell population present in a given marrow sample. Anti-AC133 MoAbs recovered approximately 75-80% of CFU-GM and CFU-Meg, but only about 30% of CFU-Mix and BFU-E. Perturbation of AC133 expression with antisense oligodeoxynucleotides (AS ODN) resulted in transient downregulation of AC133 protein on human CD34+ cells but no apparent effect on cell survival or cloning efficiency ex vivo. Finally, downregulation of MYB expression with AS ODN had no effect on the AC133 expression at either the mRNA or protein level. Based on these results, we conclude that AC133 offers no distinct advantage over CD34 or c kit as a target for immunoaffinity based isolation of primitive hematopoietic cells, that AC133 expression is not required for normal hematopoietic progenitor cell development in vitro, and finally that AC133 expression may not be MYB dependent. PMID- 10833670 TI - Coexpression of PTHrP and PTH/PTHrP receptor in a myoepithelial cell line derived from normal human breast. AB - Parathyroid hormone-related peptide (PTHrP) is the cause of humoral hipercalcaemia of malignancy syndrome (HHM). It is known that the peptide as well as its receptors are widely distributed in many normal organs and tissues, where it influences an array of diverse functions which are realized through paracrine or autocrine pathway. PTHrP is present in large amounts in lactating mammary gland but its function is not fully elucidated. In this study, production of parathyroid hormone-related peptide (PTHrP) by the Hs578Bst cell line corresponding to mammary myoepithelial cells was examined by immunocytochemistry. Using RNA extracted from these cells we analyzed expression of mRNA for PTHrP and for the PTH/PTHrP receptor by RT-PCR. The obtained results demonstrated that Hs578Bst cells produced PTHrP and synthesized mRNA for PTHrP and PTH/PTHrP type I receptor. It provides evidence that myoepithelial cells are target cells for PTHrP. The data support that PTHrP may be an important autocrine/paracrine factor, involved in the regulation of myoepithelial cell function as well as in growth and differentiation of the mammary gland. PMID- 10833671 TI - Correlation between NADPH-diaphorase and iNOS in bank vole Leydig cells in vitro and in testicular sections with the use of histochemistry and immunocytochemistry. AB - Histochemistry for NADPH-diaphorase detects an enzymatic activity associated with nitric oxide synthase while immunohistochemistry detects the nitric oxide synthase molecule. NADPH-diaphorase and inducible isoform of nitric oxide synthase in Leydig cells in vitro and in testis sections of the bank vole were demonstrated histochemically and immunocytochemically. Histochemical studies revealed localization of NADPH-diaphorase reaction product in the cytoplasm of cultured Leydig cells as well as in the interstitial area, mainly in Leydig cells and in vascular endothelium. Distribution pattern of NADPH-diaphorase was different in Leydig cell cytoplasm of individual cells. Using immunocytochemistry, the immunoreactivity for nitric oxide synthase was observed both in cultured Leydig cells and testis sections. Moreover, a co-localization of positively immunostained cells with those histochemically detected was noticed. Addition of hCG to the cultured medium or injections in vivo resulted in a small decrease in reaction intensity in Leydig cells. Treatment with N omega-nitro-L arginine methyl ester resulted in distinctly weaker reactivity of the enzymes studied which was correlated with a higher testosterone and estradiol levels in Leydig cells measured radioimmunologically. The results have indicated that nitric oxide synthase is able to act directly within the male gonad regulating androgen secretion by Leydig cells. PMID- 10833673 TI - Immunohistochemical localization of substance P in the pineal gland of the domestic pig. AB - An immunohistochemical study of the pig pineal gland was carried out using polyclonal rabbit antiserum raised against substance P (SP). The pineal glands were taken from the newborn, 21-day- and 7-month-old female pigs. Immunoreactive nerve fibers were observed in the pineal gland as well as in the posterior commissure and habenular areas. The bundles of SP-immunoreactive fibers were also seen in the subependymal layer of the pineal tissue. The single SP-immunoreactive nerve fibers and few small bundles of nerve fibers were located with equal density throughout the pineal gland, in the connective tissue septa and in the parenchyma. SP-immunoreactive cell bodies were observed in the medial habenular nucleus. The obtained results point to this nucleus as one of the central sources of SP innervation in the pig pineal gland. The study did not show any differences in the distribution and the density of SP-immunoreactive nerve fibers between newborn, 21-day- and 7 month-old pigs. PMID- 10833672 TI - Electron probe X-ray microanalysis of cultured myogenic C2C12 cells with scanning and scanning transmission electron microscopy. AB - Heterogeneity of the elemental content of myogenic C2C12 cultured cells was studied by electron probe X-ray microanalysis (EPXMA) with scanning (SEM EPXMA) and scanning transmission electron microscopy (STEM EPXMA). The best plastic substrate for growing cells was Thermanox. For STEM EPXMA, a Formvar film coated with carbon was found to be suitable substrate. The cells examined by scanning transmission electron microscopy showed great heterogeneity in their elemental content in comparison with the cells examined in the scanning electron microscope despite of an almost identical preparation procedure for EPXMA. Nevertheless the K/Na ratios obtained from both methods of EPXMA were very close (4.1 and 4.3). We conclude that the observed discrepancy in the elemental content obtained by the two methods may be due to differences in instrumentation and this must be taken into account when planning a comparative study. PMID- 10833674 TI - Proliferative activity of gastric epithelial cells in Helicobacter pylori infected children. AB - Helicobacter pylori infection has been associated with gastric carcinogenesis. Gastric epithelial cells proliferative rate is accelerated in H. pylori infected adult patients. Our study was performed to evaluate proliferative cell activity in gastric epithelium in the course of H. pylori infection in the early stage of its natural history. Gastric antral biopsy specimens were obtained from thirteen H. pylori positive and seven negative children. To assess replication rates we used nucleolar organiser regions staining with colloidal silver nitrate technique (AgNOR). The number of AgNORs per nucleus, area of single AgNOR, and the quotient of these two parameters (AgNOR content) were analysed. The mean area of AgNOR was lower in H. pylori positive than in negative children. Conversely, both the mean number of AgNOR per nucleus and AgNOR content were higher in infected than non infected subjects. These results show accelerated proliferation of gastric antral epithelial cells in the course of H. pylori infection in children. Such alteration of cell replication occurring in an initial phase of natural history of long lasting infection provides an explanation for the association between acquisition of H. pylori infection in the first years of life and the development of gastric cancer. PMID- 10833675 TI - Reductions in salivary cortisol are associated with mood improvement during relaxation training among HIV-seropositive men. AB - This study examined salivary cortisol and mood during relaxation training in 30 symptomatic, HIV+ gay men participating in a 10-week, group-based cognitive behavioral stress management intervention. Cortisol levels and mood were assessed within these sessions just before and after 45-min relaxation exercises given as part of each session. Participants also recorded their stress level and compliance with daily home relaxation practice. Presession cortisol levels decreased across the 10-week period and were related to decreases in global measures of total mood disturbance and anxious mood. Reductions in presession cortisol levels were also associated with decreases in self-reported stress level during home practice. Greater reductions in cortisol during the first three sessions were associated with more frequent relaxation practice at home. These findings suggest that salivary cortisol represents an objective neuroendocrine marker for changes in anxiety and distress observed during relaxation training in symptomatic, HIV-seropositive men. PMID- 10833676 TI - Personal, cognitive, behavioral, and demographic predictors of HIV testing and STDs in homeless women. AB - Using a multiracial sample of 621 homeless women, we tested a latent variable causal model of personal, cognitive, behavioral, and demographic predictors of two coping mediators and the outcome variables of HIV testing and return for test results and a recent STD infection. HIV testing and return were predicted by more social support, greater AIDS knowledge, greater perceived risk for AIDS, and more problem-focused coping strategies. Recent STDs were predicted by more AIDS knowledge, emotion-focused coping strategies, and risky sexual behavior and one measured variable, crack cocaine use. Emotion-focused coping strategies were predicted by drug use, less self-esteem, more social support, and greater perceived risk for AIDS. Hispanics reported less emotion-focused coping strategies than African-Americans. Predictors of problem-focused coping strategies included less drug use, more self-esteem, more social support, more AIDS knowledge, and less risky sexual behavior. African-Americans reported less problem-focused coping strategies than Latinas. Indirect effects on the outcomes mediated through coping styles are also reported. Theoretical and practical implications of results for community outreach are discussed. PMID- 10833677 TI - Behavioral treatment of insomnia: a clinical case series study. AB - There is substantial experimental evidence that behavioral treatment of insomnia produces significant clinical improvement and that treatment gains tend to be maintained over time. Less clear is whether behavioral treatment is effective as it is plied in clinical settings. In this clinical case series study, we evaluated 47 patients with primary insomnia. It was found that patients were, on average, 43% improved. This average corresponded to a 65% reduction in sleep latency, a 46% decrease in number of awakenings per night, a 48% reduction in wake time after sleep onset, and a 13% increase in total sleep time. These results suggest that behavioral treatment for insomnia is as effective in clinical settings as it is as under clinical trial conditions. PMID- 10833678 TI - Worldview and health promoting behavior: a causal model. AB - The present study investigated the manner in which Pepper's (1942) worldview theory relates to health promoting behavior. A sample of 259 subjects completed a battery of inventories measuring worldview, health promoting behavior (HPB), social class, and sex. The data were analyzed by means of structural equation modeling using the statistical program for the social sciences (SPSS) and the analysis of moment and structure (AMOS) computer programs. The results support the idea that a modest relationship exists between worldview and HPB, with organismic thinkers more likely than mechanistic thinkers to engage in HPB. There was also a slight indirect effect of sex on worldview and HPB, with women more likely to endorse an organismic worldview and therefore more likely to engage in HPB than men. No relationship was found between socioeconomic status and HPB. PMID- 10833679 TI - Hispanic adults' beliefs, attitudes, and intentions regarding the female condom. AB - The present study used the theory of planned behavior (TPB) (Ajzen, 1985) augmented by AIDS knowledge to investigate factors influencing intentions of Hispanic adults to use the female condom. A total of 146 persons (75 women and 71 men; mean age, 27 years) recruited from community-based organizations completed an anonymous survey regarding intentions to use the female condom with their main sex partner. The TPB model had greater predictive utility for women's, than for men's, female condom use intentions. For men, attitudes and norms did not predict female condom use intentions, but greater AIDS knowledge was related to lower intentions to use the female condom, above and beyond the TPB constructs. Perceived behavioral control, operationalized as self-efficacy, significantly increased the predictive utility of the TPB model for women's female condom use intentions but not for men's. Behavior change strategies to increase female condom use are discussed in light of these findings. PMID- 10833681 TI - Researchers should have talked to mothers and children. PMID- 10833680 TI - Effects of progressive relaxation and classical music on measurements of attention, relaxation, and stress responses. AB - The present experiment examined relaxation using different experimental conditions to test whether the effects of individual elements of relaxation could be measured, whether specific effects were revealed, or whether relaxation resulted from a generalized "relaxation response." Sixty-seven normal, male volunteers were exposed to a stress manipulation and then to one of two relaxation (Progressive Relaxation, Music) or control (Attention Control, Silence) conditions. Measurements of attention, relaxation, and stress responses were obtained during each phase of the experiment. All four groups exhibited similar performance on behavioral measures of attention that suggested a reduction in physiological arousal following their relaxation or control condition, as well as a decreased heart rate. Progressive Relaxation, however, resulted in the greatest effects on behavioral and self-report measures of relaxation, suggesting that cognitive cues provided by stress management techniques contribute to relaxation. PMID- 10833682 TI - More education will lead to improved end-of-life care. PMID- 10833683 TI - Psychosocial issues related to the care of adolescents with cancer. PMID- 10833684 TI - The use of amifostine to prevent xerostomia in patients receiving radiation therapy. PMID- 10833685 TI - Symptom management of the patient with a brain tumor at the end of life. PMID- 10833686 TI - Team approach for optimal patient care and satisfaction in a managed-care environment. PMID- 10833687 TI - The experience of oncology nursing: a novice staff nurse reports. PMID- 10833688 TI - Why oncology nursing? An experienced nurse replies. PMID- 10833689 TI - Tips for timely management of febrile neutropenia. PMID- 10833690 TI - Using the nursing process to implement clinical trials. PMID- 10833691 TI - Use of complementary medicine by adult patients participating in cancer clinical trials. AB - PURPOSE/OBJECTIVES: To document the prevalence, demographic correlates, patterns of use, and beliefs about complementary and alternative medicine (CAM) therapies of adult patients enrolled in National Cancer Institute (NCI) clinical trials. DESIGN: Prospective, cross-sectional, descriptive survey. SETTING: W.G. Magnuson Clinical Center of the National Institutes of Health in Bethesda, MD. SAMPLE: Convenience sample of 100 English-speaking, adult patients with cancer admitted to intramural clinical trials. METHODS: A standardized, 99-item questionnaire assessing use of CAM therapies pre- and postcancer diagnosis was administered by face-to-face interview. MAIN RESEARCH VARIABLES: Use of CAM therapies, beliefs, communication with physician. FINDINGS: 63% used at least one CAM therapy, with an average use of two therapies per patient. Men were significantly less likely to use a therapy than women; women were more likely to use numerous therapies. Cancer diagnosis seems to have had no influence overall on the frequency of use of CAM therapies. The major reasons stated for CAM use were for treatment-related medical conditions as well as depression, anxiety, and insomnia. The most frequently reported therapies were spiritual, relaxation, imagery, exercise, lifestyle diet (e.g., macrobiotic, vegetarian), and nutritional supplementation. Patients unanimously believed that these complementary therapies helped to improve their quality of life through more effective coping with stress, decreasing the discomforts of treatment and illness, and giving them a sense of control. CONCLUSIONS: Patients with cancer use various complementary therapies to cope with their disease and the rigors of clinical trials. Women and those with higher educational backgrounds were more frequent users. IMPLICATIONS FOR NURSING PRACTICE: Nurses who provide care to subjects of biomedical research have an opportunity and responsibility regarding their patients' use of CAM therapies. Nurses may use in-house resources to help evaluate subjects' use of a CAM modality or to provide quality-of-life therapies such as relaxation, imagery, or healing touch. Discussing these health practices in a nonjudgmental manner adds to the assessment of patients' coping skills and ability to make decisions about their health care. PMID- 10833692 TI - Demands of illness in people treated for colorectal cancer. AB - PURPOSE/OBJECTIVES: To describe the most common and most intense demands of illness in people with colorectal cancer. DESIGN: Descriptive, comparative. SETTING: People with colorectal cancer were accrued through weekly online computer postings. SAMPLE: 121 people (68 men and 53 women) from 35 states (n = 106) and six other countries (n = 15) who were treated for colon, rectal, or anal cancer. METHODS: Mailed questionnaires included a 125-item, self-report instrument with seven subscales (Demands of Illness Inventory (DOII) and a 14 item demographic form. MAIN RESEARCH VARIABLES: Demands of illness, time since treatment, perception of illness state, activity level, and age. FINDINGS: Demands of illness were greatest in the personal meaning domain, with more than 93% of subjects reporting that they thought about the value of life and how long they might live. More than 85% reported uncertainty in six similar areas. The 10 most intense demands were predominately psychosocial and existential concerns. Respondents in the youngest age group (< 45 years) reported significantly greater demands in six of seven domains of the DOII than the middle and older groups. Three variables (time since treatment, perception of illness, activity level) accounted for 45% of the variance in DOII scores. CONCLUSIONS: Colorectal cancer imposed significant psychosocial and existential concerns on respondents, especially the youngest age group. IMPLICATIONS FOR NURSING PRACTICE: Nurses should address these concerns with patients while continuing to identify and scientifically test creative strategies to minimize psychosocial morbidity. PMID- 10833693 TI - Recent developments in understanding the immune response to human papilloma virus infection and cervical neoplasia. AB - PURPOSE/OBJECTIVES: To provide an overview of the immune system and describe the systemic and local immune response to human papilloma virus (HPV) infection in the cervix and to the development of cervical dysplasia. DATA SOURCES: Historical and current medical and nursing literature, current gynecologic oncology texts. DATA SYNTHESIS: The immune system provides protection against a wide variety of pathogens and plays a role in the response of the body to neoplastic cells. The immune system orchestrates the functions of various immune cells and proteins to fight against invading pathogens. Mucosal immunity is one of the key functions of the immune system and has unique features. Humoral and cytotoxic responses in the cervix and in the peripheral blood are seen during clearance of an HPV infection. CONCLUSIONS: The functional status of the immune system is associated closely with the development of cervical dysplasia and cancer in women with HPV infection. Clinicians must assist women in maintaining proper functioning of immune responses. IMPLICATIONS FOR NURSING PRACTICE: A better understanding of local and systemic immune responses to HPV infection may help nurses who provide care to women with cervical disease and women at high risk for cervical cancer to deliver better care and clearer information to patients. PMID- 10833694 TI - Work-site cancer screening: a Latino case study. AB - PURPOSE/OBJECTIVES: To develop a work-site cancer education and detection program targeted for Latino women and evaluate the effectiveness of the program in a firm with a majority of employees from Latino backgrounds. DESIGN: Program implementation and evaluation. SETTING: A Latino-owned industrial firm in Detroit, MI. SAMPLE: 560 of 857 female employees. Among the 202 women who were eligible for mammography, 142 participated in the screening. METHODS: Education and mammography were offered to women employees of Mexican Industries in Michigan. Education programs and mammograms were conducted at the work site. The programs were offered in English and Spanish. MAIN OUTCOME MEASURES: Participation in education and mammography and evaluation of program components. FINDINGS: Pre- and post-tests of knowledge indicated increases in the understanding of breast cancer. Evaluation of the educational program indicated high ratings of the presentations and materials. CONCLUSIONS: Work-site cancer education and detection programs are cost effective in firms where the majority of employees are from one ethnic/racial group and where the programs take into consideration the cultural background of the employees. IMPLICATIONS FOR NURSING PRACTICE: Cancer education and detection programs should consider the possibility of work-site interventions as well as the ethnic/racial background of employees during program development. PMID- 10833695 TI - Cancer-related depression: Part I--Neurologic alterations and cognitive behavioral therapy. AB - PURPOSE/OBJECTIVES: To describe neurologic and cognitive alterations underlying symptoms of depression and to explore cognitive-behavioral approaches to promoting recovery from cancer-related depression. DATA SOURCES: Published literature, unpublished raw data, and clinical observations. DATA SYNTHESIS: Depression is a progressive condition that is most responsive to treatment in its earliest stages because of the progressive nature of alterations in neurologic circuits and neurotransmitters. Aggressive screening and management using cognitive-behavioral therapy (CBT) techniques can promote recovery from cancer related depression and improve patients' quality of life. Application of CBT techniques to patient environments also holds promise of relieving and preventing depression. IMPLICATIONS FOR NURSING PRACTICE: By placing more emphasis on screening for cancer-related depression among newly diagnosed and treated patients, oncology nurses can expedite treatment of cancer-related depression. Working within psychiatric liaison teams or guidelines for routine psychiatric care, oncology nurses can promote recovery and create therapeutic environments that are conducive to promoting patients' mental health along the cancer trajectory. PMID- 10833696 TI - Telephone therapy for patients with breast cancer. AB - PURPOSE/OBJECTIVES: To test the value of telephone-administered cognitive behavioral therapy in a study of patients with breast cancer. DESIGN: Women were assigned randomly to a therapy group or an assessment-only control group. SETTING: A tertiary cancer treatment center serving rural areas of North Dakota and Minnesota. SAMPLE: Women were recruited within three to four months of stage I (n = 27) or stage II (n = 26) breast cancer diagnosis. Age ranged from 30-82 (mean = 51.5 years). Most participants (n = 35) underwent a modified radical mastectomy; 17 underwent a lumpectomy. METHODS: Therapy involved 10 30-minute (or less) telephone sessions. Data that were collected from mailed questionnaires included psychological distress (Profile of Mood States), perceived stress, coping (Coping Response Indices-Revised), quality of life (Medical Outcome Scale), and satisfaction with therapy. Measures were completed at baseline and at 4- and 10-month follow-up intervals. MAIN RESEARCH VARIABLES: Telephone therapy, stress, coping, and quality of life. FINDINGS: With time, women in the therapy and control groups reported reduced stress and improved quality of life. However, significant reductions in some kinds of distress (anxiety, anger, depression, and confusion) were not observed. Most therapy participants liked the telephone treatment sessions but showed only modest improvement (less anxiety and confusion) compared with women in the control group. CONCLUSIONS: Most patients reported being comfortable with the telephone therapy and said that they felt better as a result of it. However, the outcome data showed that telephone therapy -as carried out in this study--produced only modest benefits. Researchers need to consider who is best for delivering such therapy. IMPLICATIONS FOR NURSING PRACTICE: Providing telephone therapy to patients with breast cancer has potential benefits, and nurses may be the appropriate professionals to administer the therapy. PMID- 10833697 TI - The experience of donating bone marrow to a relative. AB - PURPOSE/OBJECTIVES: To describe and understand the experience of donating bone marrow to a relative. DESIGN: Exploratory, descriptive, and qualitative. SETTING: An urban comprehensive cancer center. SAMPLE: Twelve donors were interviewed- eight women and four men. The average age was 47, and the average length of time since donation was nine months. At the time of the interview, seven recipients were living and five had died. METHODS: Open-ended, face-to-face, or telephone interviews were conducted within one year of bone marrow donation using an interview guide. MAIN RESEARCH VARIABLES: Psychosocial consequences of donating bone marrow to a relative. FINDINGS: The main theme was Doing What It Takes When a Family Member Has Cancer. This goal guided the decision-making process, psychological responses, and family relationships once transplantation was necessary. Six subthemes emerged: Dealing With the Donation Procedure, Informational Needs, Psychological Impact of the Donor Role, Managing Family Relationships, Monitoring One's Own Health Before Donation, and Adjusting to the Transplant Recipient's Medical Condition. CONCLUSIONS: Subjects expressed little or no reluctance to donate bone marrow, and all would repeat the experience. Subjects felt deep personal satisfaction and gratitude for an opportunity to donate. Stressful aspects of the experience related to unanticipated pain after the procedure, negative transplant outcomes, and relationships with the bone marrow recipients' family. IMPLICATIONS FOR NURSING PRACTICE: Predonation assessment should identify donors' need for information as well as types and sources of information, how donors and families have coped with previous crises, and discussions of potential negative transplantation outcomes and impact on donors. Donors' coping and adjustment should be monitored throughout the transplantation process, particularly when coping problems or potential problems are identified or with negative transplantation outcomes. Formal donor support programs should be considered. PMID- 10833698 TI - Device follow-up in the age of automaticity. PMID- 10833699 TI - Regional differences in arrhythmogenic aftereffects of high intensity DC stimulation in the ventricles. AB - Regional differences of the aftereffects of high intensity DC stimulation were investigated in isolated rabbit hearts stained with a voltage-sensitive dye (di-4 ANEPPS). Optical action potential signals were recorded from the epicardial surface of the right and left ventricular free wall (RVep, LVep) and from the right endocardial surface of the interventricular septum (IVS). Ten-millisecond monophasic DC stimulation (S2, 20-120 V) was applied to the signal recording spots during the early plateau phase of the action potential induced by basic stimuli (S1, 2.5 Hz). There was a linear relationship between S2 voltage and the S2 field intensity (FI). S2 caused postshock additional depolarization, giving rise to a prolongation of the shocked action potential. With S2 > or = 40 V (FI > or = approximately 20 V/cm), terminal repolarization of action potential was inhibited, and subsequent postshock S1 action potentials for 1-5 minutes were characterized by a decrease in the maximum diastolic potential and a decrease in the amplitude and a slowing of their upstroke phase. The higher the S2 voltage, the larger the aftereffects. The changes in postshock action potential configuration in RVep were significantly greater than those observed in LVep and IVS when compared at the same levels of S2 intensity. In RVep, 12 of 20 shocks of 120 V resulted in a prolonged refractoriness to S1 (> 1 s), and the arrest was often followed by oscillation of membrane potential. Ventricular tachycardia or fibrillation ensued from the oscillation in five cases. No such long arrest or serious arrhythmias were elicited in LVep and IVS. These results suggest that RVep is more susceptible than LVep and IVS for arrhythmogenic aftereffects of high intensity DC stimulation. PMID- 10833700 TI - Voltage dependence of ICD lead polarization and the effect of iridium oxide coating. AB - Nonthoracotomy leads (NTLs) with an iridium oxide (IROX) coating exhibit lower defibrillation thresholds (DFTs) than uncoated NTLs. We tested whether adding an IROX coating to an active pectoral can would influence defibrillation efficacy. However, the primary purpose of this study was to examine the impedance changes that occur at different voltages for uncoated titanium NTLs and identical NTLs with an IROX coating. We studied anesthetized pigs with an NTL placed in the right ventricle and coupled this to an active pectoral can. Biphasic waveform DFTs were obtained for the four NTLs and can combinations: uncoated NTL and uncoated can, uncoated NTL and IROX can, IROX NTL and uncoated can, and IROX NTL and IROX can. The respective energy DFTs were: 23.6 +/- 6.9, 24.1 +/- 6.7, 21.3 +/- 6.0, and 21.4 +/- 7.0 J. The IROX NTL DFTs were significantly lower (P < 0.05) than the uncoated NTL DFTs (either can), confirming our previous study. We then used a low tilt monophasic waveform to assess impedance changes. The impedance rise for each NTL/can combination was measured at 50, 100, 300, and 700 V. Comparisons of impedance changes between voltage levels showed that the impedance rise was inversely related to voltage and was greatest with uncoated NTLs. The IROX coating of the NTL reduced the impedance rise at all shock voltages, but was particularly beneficial at the lower voltages. No advantage was seen when the pectoral can was coated with IROX regardless of which NTL was used. Our results suggest that low voltage applications, such as atrial defibrillation, would benefit most from the IROX-coated NTL, and further studies are warranted in this area. PMID- 10833701 TI - Is automatic mode switching effective for atrial arrhythmias occurring at different rates? A study of the efficacy of automatic mode and rate switching to simulated atrial arrhythmias by chest wall stimulation. AB - Automatic mode switching (AMS) is a useful means to avoid rapid ventricular response during atrial fibrillation (AF), but AMS cannot occur if the detected atrial rate during AF is below the mode switching criteria. This may be the result of antiarrhythmic medications, or when the atrial events fall within the atrial blanking period, or if the atrial amplitudes during AF are too small to be sensed. We hypothesize that the addition of an automatic rate switching (ARS) algorithm may complement AMS response during AF with different detected atrial rates. We studied the Marathon DDDR pacemaker (Model 294-09, Intermedics Inc.) with the AMS and ARS algorithms that are independently programmable but can also operate in combination. AF sensed above the AMS rate (160 beats/min) will lead to VDIR pacing, whereas AF below AMS rate will be tracked at an interim rate as dictate by the ARS, at a ventricular response that is 20 beats/min above the sensor indicated rate. Atrial tachyarrhythmias were simulated by chest wall stimulation (CWS). CWS was applied to 33 patients (16 men, 17 women, mean age 69 +/- 11 years) with a Marathon DDDR pacemaker using an external pacer to simulate AF occurring at two rate levels: above the AMS rate (programmed at 160 beats/min) at 180 beats/min and below the AMS rate at 120 beats/min. The maximum, minimum, and mean ventricular rates during CWS in DDDR mode with AMS alone, ARS alone, and their combination were compared. During CWS at 120 beats/min, the AMS plus ARS setting showed a mean ventricular rate of 79 +/- 3 beats/min and 124 +/- 14 beats/min in the AMS setting alone (P < 0.01). With CWS at 180 beats/min, the mean ventricular rate in the AMS plus ARS setting compared to the AMS setting alone was not significantly different. However, the variation in ventricular pacing rate was 7 +/- 14 beats/min in the AMS plus ARS setting and 40 +/- 42 beats/min in the AMS setting (P < 0.05). In conclusion, AMS is effective for simulated atrial tachyarrhythmias sensed above the AMS rate. Combined AMS with ARS is useful to handle simulated atrial tachyarrhythmia at a slower rate and to avoid rate fluctuation during AMS. There is also a possibility that this can be applied to the naturally occurring atrial tachyarrhythmias. PMID- 10833702 TI - Atrial electrophysiological effects of ibutilide infusion in humans. AB - Patients with persistent atrial fibrillation (AF) who respond to ibutilide infusion usually convert to sinus rhythm within 1 hour. However, little is known about the magnitude and time course of the drug's atrial electrophysiological effects. In the current study, the frequency content of the fibrillatory baseline on the ECG was used to quantify the effects of ibutilide infusion on AF. Nineteen patients (age 56 +/- 16 years) in persistent AF were studied. Nine of these were chronically treated with antiarrhythmic drugs. All subjects received ibutilide (1 mg i.v.) over 10 minutes and a second dose 10 minutes later as needed. An ECG was recorded and digitized throughout each of the 20 sessions (one patient had two separate cardioversions). A signal processing technique was then used to quantify the average rate of the fibrillatory baseline. After attenuating the QRS and T waves, the ECG was subjected to Fourier transformation. The average rate of fibrillatory activity was defined as the frequency corresponding to the peak power of this spectrum. The fibrillation rate declined by 20 +/- 12% during the initial dose of ibutilide. This effect was rapid with t1/2 = 4.2 minutes. Ibutilide induced slowing of AF was more intense and more rapid in patients who converted to sinus rhythm than in those who did not (25 +/- 5% vs 18 +/- 14% drop in rate, t1/2 3.4 minutes vs 6.3 minutes, P = 0.002). Ibutilide induced slowing of fibrillatory activity occurs rapidly and approaches steady state before the end of a 10-minute infusion. Although interaction with other antiarrhythmic drugs may have confounded the results, the speed and intensity of slowing correlated with conversion. These observations suggest that additional studies are warranted to determine if ibutilide dosing regiments can be optimized. PMID- 10833703 TI - An association between anxiety and neurocardiogenic syncope during head-up tilt table testing. AB - To study the association between anxiety and neurocardiogenic syncope as determined by head-up tilt table testing (HUT) in men and women with presyncope or syncope, patients with unexplained syncope or presyncope undergoing HUT were asked to complete the Burns Anxiety Inventory (BAI), a validated inventory of 33 questions with responses graded from 0 to 3. HUT consisted of a 30-minute tilt to 60 degrees, which if negative, was repeated with an isoproterenol infusion. A positive HUT was defined as symptomatic hypotension and/or bradycardia. Of the 66 patients who completed the BAI and underwent HUT, 33 were men and 33 were women. The mean age was 57 +/- 18 years (17-91 years). Patients with a positive HUT had a higher BAI score than those with a negative HUT (22 +/- 12 vs 14 +/- 13, P = 0.017). This association was stronger in women with a BAI score of 24 +/- 11 in those with a positive HUT versus 13 +/- 8 in those with a negative HUT (P = 0.005). In contrast, the mean BAI score for men with a positive HUT was 19 +/- 13, as compared to 15 +/- 16 for a negative HUT (P = 0.5). In conclusion, the present study demonstrates a statistical association between anxiety (as determined by BAI) and HUT result. Gender-based analysis revealed a more statistically significant relationship between anxiety and HUT outcome for women as compared to men. PMID- 10833704 TI - Factors affecting the frequency of subcutaneous lead usage in implantable defibrillators. AB - Subcutaneous leads (SQ) add complexity to the defibrillation system and the implant procedure. New low output devices might increase the requirement for SQ arrays, although this might be offset by the effects of active can and biphasic technology. This study sought to assess the impact of these technologies on SQ lead usage, and to determine if clinical variables could predict the need for an SQ lead. Patients receiving nonthoracotomy systems (n = 554) at our institution underwent step-down-to-failure DFT testing with implant criteria of a 10-J safety margin. SQ leads were used only after several endovascular configurations failed. Use of biphasic waveforms significantly lowered the frequency of use of SQ leads from 48% to 3.7% (P < 0.000001). SQ leads were required in 4.4% of patients with cold can devices and 2.6% of patients with active can devices (P = NS). There was no increase in SQ lead usage with low energy (< 30-J delivered energy) devices. Clinical variables (including EF, heart disease, arrhythmia, and prior bypass) did not predict the need for an SQ lead. The implant DFT using SQ arrays (14.5 +/ 6.5 J) was not significantly lower than that for SQ patches (16.6 + 6.0 J). We conclude that biphasic waveforms significantly reduce the need for SQ leads. Despite this reduction, 3.7% of implants still use an SQ lead to achieve adequate safety margins. The introduction of lower output devices has not increased the need for SQ leads, and when an SQ lead is required, there is not a significant difference in the implant DFT of patches versus arrays. Clinical variables cannot predict which patients require SQ leads. PMID- 10833705 TI - Age effect on phase relations between respiratory oscillations of the RR interval and systolic pressure. AB - Spectral analysis may allow the evaluation of (baroreflex) gain and phase between the RR interval and systolic pressure oscillations synchronous with respiration but, unlike baroreflex gain, the determinants of phase are not completely understood. We evaluated the correlates of spectral phase in 92 healthy subjects (44 men) aged 10-80 years. To do so, the cardiorespiratory signals during paced breathing at 16 breaths/min were continuously recorded and analyzed. In addition, respiratory sinus arrhythmia and baroreflex gain (two indices of cardiac vagal activity) and phase were calculated by using an autoregressive spectral technique. At univariate analysis, the phase correlated with age (r = 0.48, P < 0.001), the RR interval (r = 0.32, P < 0.01), respiratory sinus arrhythmia (r = 0.3, P < 0.01), baroreflex gain (r = -0.29, P < 0.01), and body mass index (r = 0.25, P < 0.05). At multivariate analysis, age was the most important physiological correlate of phase, accounting for 23% of interindividual phase variation. Cardiac vagal activity measures (which were higher in women than men) and the RR interval were also significant independent correlates of phase. We conclude that in addition to the RR interval and cardiac vagal activity, age has a significant impact on the phase relationship between respiratory related oscillations of the RR interval and systolic blood pressure. This spectral measure may contain additional information concerning the mechanisms that influence cardiovascular rhythms. PMID- 10833706 TI - Infarct related artery patency: relation to serial electropharmacological studies and outcome in patients with previous myocardial infarction and ventricular tachyarrhythmias. AB - Evidence suggests that infarct related artery (IRA) patency may improve survival after acute myocardial infarction, which is thought to be partially due to a lower incidence of malignant ventricular tachyarrhythmias. However, little is known about the effect of IRA patency on antiarrhythmic drug response and long term outcome in patients with previous infarction who already experienced sustained ventricular tachyarrhythmias. A total of 152 patients with remote myocardial infarction and documented ventricular tachycardia (VT) or ventricular fibrillation (VF) underwent coronary angiography and programmed ventricular stimulation before and after oral administration of d,l-sotalol (240-640 mg/day). D,l-sotalol suppressed inducibility of VT/VF in 37 (25.2%) patients. The IRA was patent in 38.1% of all patients. There was no significant difference in the frequency of drug response between patients with patent or occluded IRAs (26.8% vs 24.2%, P = 0.87). In patients with a patent IRA, d,l-sotalol tended to be more effective in the absence of a left ventricular aneurysm, although this difference did not reach statistical significance (P = 0.38). Ejection fraction and collateral blood flow had no effect on drug response in the presence or absence of IRA patency. During follow-up (13.0 +/- 19.9 months) of 29 patients discharged on oral d,l-sotalol, 3 patients experienced symptomatic VT and 4 sudden death. Arrhythmia recurrence and death of all cause (n = 6) and cardiac death (n = 4) were independent of IRA patency status. IRA patency had no effect on short-term drug response to d,l-sotalol in patients with remote myocardial infarction and documented VT/VF. Long-term outcome of patients with sustained ventricular tachyarrhythmias is independent of IRA patency status. In contrast to a previous report, outcome of electropharmacological testing was not predicted by the patency of the IRA. PMID- 10833707 TI - Age dependency of sensing performance and AV synchrony in single lead VDD pacing. AB - Implantation of single lead VDD pacemakers is an established therapeutic option in patients with AV block and normal sinus node function. However, related to occurrence of sinus node disease and atrial undersensing, there is concern whether VDD devices are appropriate in physically active young patients. Two hundred thirty-two consecutive patients with isolated AV block and VDD pacemakers were investigated. This population was subdivided into quartiles of 58 patients according to age at time of inclusion: 26.2-59.4 years (group A), 59.5-70.1 years (group B), 70.2-81.0 years (group C), and 81.1-92.5 years (group D). Follow-up visits included pacemaker telemetry, Holter monitoring, and exercise testing. Patients were visited at 2 and 12 weeks after implantation thereafter followed by 6-month intervals. Mean follow-up period was 35 +/- 14 months. Three months after implantation, atrial sensing threshold was significantly higher in young patients: 1.18 +/- 0.58 mV (group A) versus 0.79 +/- 0.35 mV (group B), 0.68 +/- 0.33 mV (group C), and 0.60 +/- 0.25 mV (group D), P < 0.001 for comparison of group A to all other groups. Atrial undersensing was observed less frequently in young patients: 6.9% (group A) versus 17.2% (group B), 24.1% (group C), and 27.6% (group D), P = 0.025 for intergroup comparisons. Sinus node dysfunction did not occur in group A. Atrial arrhythmias and loss of AV synchronized pacing mode occurred rarely in young patients: 0.6% (0.4%) per year in group A versus 1.3% (1.3%) in group B, 3.9% (3.4%) in group C, and 5.7% (7.4%) per year in group D, P < 0.01 for intergroup comparisons. Our data show good atrial sensing performance, low incidence of sinus node dysfunction, and few atrial arrhythmias in young patients with VDD pacing for AV block. Thus, single lead VDD pacing can be recommended particularly for young patients with AV block. PMID- 10833708 TI - Successful treatment of idiopathic left ventricular outflow tract tachycardia by catheter ablation or minimally invasive surgical cryoablation. AB - Idiopathic right ventricular outflow tract tachycardia is readily amenable to radiofrequency catheter ablation. However, treatment modalities for left ventricular outflow tract tachycardia are not well defined. Out of 37 patients with idiopathic outflow tract tachycardia referred for catheter ablation, in 3 patients tachycardia originated from the left ventricular outflow tract. On the surface ECG, all left ventricular tachycardias exhibited an inferior axis with a predominant negative QRS complex in lead I. Heart rate during tachycardia ranged from 115 to 170 beats/min. During electrophysiological testing, 1 patient had inducible tachycardia on orciprenaline challenge, 1 patient had inducible tachycardia at baseline, and 1 patient had incessant tachycardia. In two patients, earliest ventricular activation was recorded from the endocardial left ventricular outflow tract at an anterolateral and an anterior site, respectively. A distinct high frequency spike preceded the QRS onset by 66/78 ms. Application of radiofrequency energy successfully eliminated tachycardia at these sites. In one patient, tachycardia originated from the epicardial left ventricular outflow tract. Mapping of the anterior interventricular vein revealed a fractionated low amplitude signal occurring 46 ms before QRS onset. After failure of catheter ablation from the corresponding endocardial site, successful minimally invasive surgical focal cryoablation of the epicardial target region was performed. During a follow-up period ranging from 7 to 12 months, all patients remained free of tachycardia. In conclusion, ventricular tachycardia arising from the left ventricular outflow tract may require endo- and epicardial mapping. Successful treatment is achieved by radiofrequency catheter ablation or minimally invasive surgical cryoablation. PMID- 10833709 TI - Diagnosis and management of inadvertently placed pacing and ICD leads in the left ventricle: a multicenter experience and review of the literature. AB - Three patients from different centers with pacemaker or ICD leads endocardially implanted in the left ventricle are described. All leads, two ventricular pacing leads and one ICD lead, were inserted through a patent foramen ovale or an atrial septum defect. The diagnosis was made 9 months, 14 months, and 16 years, respectively, after implantation. All patients had right bundle branch block configuration during ventricular pacing. Chest X ray was suggestive of a left sided positioned lead except in the ICD patient. Diagnosis was confirmed with echocardiography in all patients. One patient with a ventricular pacing lead presented with a transient ischemic attack at 1-month postimplantation. During surgical repair of the atrial septum defect 14 months later, the lead was extracted and thrombus was attached to the lead despite therapy with aspirin. The other patients were asymptomatic without anticoagulation (9 months and 16 years after implant). No thrombus was present on the ICD lead at the time of the cardiac transplantation in one patient. We reviewed 27 patients with permanent leads described in the literature. Ten patients experienced thromboembolic complications, including three of ten patients on antiplatelet therapy. The lead was removed in six patients, anticoagulation with warfarin was effective for secondary prevention in the four remaining patients. In the asymptomatic patients, the lead was removed in five patients. In the remaining patients, 1 patient was on warfarin, 2 were on antiplatelet therapy, and in 3 patients the medication was unknown. After malposition was diagnosed, three additional patients were treated with warfarin. In conclusion, if timely removal of a malpositioned lead in the left ventricle is not preformed, lifelong anticoagulation with warfarin can be recommended as the first choice therapy and lead extraction reserved in case of failure or during concomitant surgery. PMID- 10833710 TI - Use of a single coil transvenous electrode with an abdominally placed implantable cardioverter defibrillator in children. AB - While transvenous defibrillator electrode placement avoiding a thoracotomy is preferable, electrode size, a large intercoil spacing, and the need for subclavicular device placement preclude this approach in most children. We investigated a single RV coil to an abdominally placed active can ICD device. Five children ages 8-16 years (weight 21-50 kg, mean 35 kg) underwent ICD placement. Placement of a single coil Medtronic model 6932 or 6943 electrode was performed via the left subclavian vein approach and the electrode positioned in the RV apex with the coil lying along the RV diaphragmatic surface. The ICD (Medtronic Micro Jewel II model 7223 Cx) was implanted in a left abdominal pocket with the lead tunneled from the infraclavicular region to the pocket. Implant DFTs were < or = 15 J using a biphasic waveform. DFTs rechecked within 3-month postimplant were unchanged. Lead impedance at implant ranged from 38 to 56 omega, mean 51 omega. Follow-up was 3-21 months (total 82 months) with no electrode dislodgment, lead fractures, or inappropriate discharges. Two of the five patients have had successful appropriate ICD discharges. Transvenous ICD electrode placement can be performed in children as small as 20 kg with the device implanted in a cosmetically acceptable abdominal pocket that is well tolerated. Excellent DFTs can be achieved. This approach avoids a thoracotomy in all but the smallest child, does not require subclavicular placement of the device, and avoids use of a second intravascular coil. PMID- 10833711 TI - Prevalence of late potentials in a sample of 487 healthy, middle-aged men from southwestern France. AB - Late potentials (LPS) have been shown to be predictive of ventricular tachycardia in coronary artery disease subjects, but the prevalence of LP in the general population is not as well-known. The study included 487 men without a history of cardiovascular disease (aged 50-59) living in Southwestern France. Standard averaged high resolution electrocardiogram was performed using Butterworth filtering at 40-250 Hz. LPS were defined as two or more of the following criteria: QRS > 114 ms, duration of the low amplitude signals in the terminal portion of QRS > 38 ms, root mean square (RMS) voltage of the last 40 ms < 20 microV. The mean QRS duration was 97 +/- 12 ms (mean +/- SD), duration of the low amplitude signals in the terminal portion of QRS was 32 +/- 10 ms, and RMS voltage in the last 40 ms was 39 +/- 27 microV. Eight percent of subjects (95% confidence interval [CI]: 6%-11%) had a QRS duration > 114 ms; 22% (95% CI: 18% 26%) had a duration of low amplitude signals > 38 ms, and 25% (95% CI: 22%-29%) had RMS voltage in the last 40 ms < 20 microV. Finally, the prevalence of LP was 21% (95% CI: 18%-25%). In conclusion, according to commonly used criteria, the prevalence of LP in this healthy population of middle-aged men is 21%, close to the values found in the literature for myocardial infarction patients. Those findings indicate the need for reconsidering the definition of LPS. PMID- 10833712 TI - Therapy of atrial fibrillation: rhythm control versus rate control. PMID- 10833713 TI - Two types of tachycardia, one lesion. PMID- 10833714 TI - Transthoracic pacing in a very low birth weight infant with congenital complete atrioventricular block. AB - We report our experience of pacemaker treatment in a premature infant of 830 g with congenital complete atrioventricular block due to maternal Sjogren's Syndrome. The infant was delivered by cesarean section at an estimated gestational age of 26 weeks because of fetal bradycardia, decreasing fetal movements, and hydrops. Immediate postnatal transesophageal ventricular pacing was not successful, whereas transthoracic pacing with self-adhesive patch electrodes adapted to body size resulted in an effective increase of the infant's heart rate until operative application of temporary epimyocardial pacing wires allowed external stimulation of the heart. PMID- 10833715 TI - ICD hardware failure associated with multiple internal shocks. AB - The delivery of 37 shocks by an ICD within 20 minutes, in response to T wave oversensing during atrial flutter, resulted in several manifestations of undesirable device behavior. The generator reverted to backup mode, and disabled automatic capacitor reformation, therapy delivery, and automatic gain control. Postexplant analysis of the device revealed damage to the high voltage output section of the circuitry consistent with excessive electrical stress. In rare circumstances, multiple internal discharges can result in serious clinical anomalies in ICD behavior, and possibly in an increase in susceptibility to circuitry damage. PMID- 10833716 TI - Radiofrequency ablation of an accessory pathway in a surgically created atrioventricular Fontan anastomosis: case report and review of previous published cases. AB - Supraventricular tachyarrhythmias following the Fontan procedure can be life threatening. Though most are commonly due to intraatrial reentry, orthodromic reentrant tachycardia may also be present. Atrioventricular accessory pathways may develop across suture lines following right atrial to right ventricular anastomosis in patients with tricuspid atresia. We report a case of a patient who underwent this type of Fontan who developed orthodromic reentrant tachycardia and heart failure. An electrophysiological study revealed the presence of an atrioventricular accessory pathway traversing the Fontan anastomosis suture line. Successful radiofrequency ablation of the accessory pathway led to control of the tachyarrhythmia and improvement of heart failure. PMID- 10833717 TI - Block of the lower interatrial connections: insight into the sources of electrocardiographic diversities in common type atrial flutter. AB - Whether the conduction disturbances of the interatrial connections play a role in the genesis of ECG variants of atrial flutter is almost completely unknown. We present a patient with typical counterclockwise atrial flutter in whom the ablation of the coronary sinus (CS) area during ongoing atrial flutter produced significant ECG changes without alterations in the activation sequence within the right atrium (RA). This case highlights the possible role of alterations of the interatrial connections in the genesis of atypical ECG manifestations of common type atrial flutter. PMID- 10833718 TI - Pathological findings of the isthmus between the inferior vena cava and tricuspid annulus ablated by radiofrequency application. AB - Anatomically guided radiofrequency ablation for the treatment of atrial flutter was performed in a 41-year-old man with interstitial pneumonia. He died of respiratory failure 2 months after ablation, and an autopsy was performed. The whole layer of the ablation site showed a transluminal fibrosis. PMID- 10833719 TI - Prediction of optimal atrioventricular delay in patients with implanted DDD pacemakers. PMID- 10833720 TI - Optimal cardiac pacing after heart transplantation. PMID- 10833721 TI - Minimally invasive open heart surgical techniques. PMID- 10833722 TI - Proliferation of dual-chamber antibradycardia pacemakers capable of changing pacing modes automatically. PMID- 10833723 TI - Auditory backward-masking performance by children who stutter and its relation to dysfluency rate. AB - The fluency of people who stutter is affected markedly when auditory feedback is altered, suggesting that stuttering may be associated with hearing. Peripheral hearing problems, however, are no more common in people who stutter than in those who do not. Performance was investigated in a task that involves central auditory processing (backward masking). Children who stuttered had deficits in backward masking (indicated by higher thresholds) compared with a group of fluent control children. The backward-masking thresholds were positively correlated with frequency of stuttering. PMID- 10833724 TI - Suggesting strategies improves creative visual synthesis. AB - An experiment assessed whether a figural or an interpretative strategy can enhance creative visual synthesis. 45 undergraduates were presented a set of simple figures and asked to imagine combining them to obtain a whole pattern corresponding to a creative product. In the figurative condition participants were instructed to combine figures in unusual ways; in the interpretative condition they were induced to look for unusual meanings embedded in the combinations; in the control condition no strategy was suggested. Results showed that certain strategies induced a more flexible visual synthesis. PMID- 10833725 TI - Imagix: multimedia software for evaluating the vividness of movement-imagery. AB - This article presents a new software, Imagix, designed for the evaluation of the vividness of movement-imagery. This multimedia test is comprised of 18 movements in six different psychomotor categories whose vividness of imagery is to be appraised. Imagix has the advantage of offering an evaluation of the vividness of imagery in a normative way. The participant chooses among four films displaying a vividness gradient, which includes contrast and luminosity. The participant is first asked to visualize a movement verbally suggested. The participant is then asked to evaluate the vividness of this mental image by choosing on a computer screen the movie that best corresponds to the vividness of the movement-imagery. The first versions of this freeware (in English and French) are currently being validated. PMID- 10833726 TI - Left-handedness and inheritance of bronchial asthma. AB - A statistically significant difference in mean laterality quotients of 15 asthmatic and 106 nonasthmatic children showed a coexistence of left-handedness and asthma early in life. It is suggested that this phenomenon is not only a fetal event as the Geschwind-Galaburda hypothesis claims but it is mainly based on maternal line inheritance. PMID- 10833727 TI - The Bender-Gestalt test in an Italian sample: an analysis of Koppitz's Developmental Bender Scoring System deviations. AB - This study extended the research of the psychometric characteristics of Koppitz's 1963/1975 Developmental Scoring System of the Bender-Gestalt test. Attention was paid to relations among the 7 deviations. The test was administered by licensed psychologists to 1,065 white children, aged from 3 yr., 6 mo. to 11 yr., 5 mo., enrolled in the regular education track of kindergarten and elementary school in Italy. PMID- 10833728 TI - Anhedonia, asthenia, and depression. AB - In a sample of 264 university students, anhedonia scores were not associated with scores for severity of fatigue. PMID- 10833729 TI - Structure of the Mini-Mental State Examination: a comparison between normal and clinical subjects. AB - The Mini-Mental State Examination was administered to a sample of 133 Italians ranging from 60 to 80 years of age. From a Principal Components Analysis three factors were retained, being results of both varimax and oblimin rotations. These were labeled (1) Memory, (2) Verbal Comprehension, and (3) Attention. These factors are similar to those obtained with normal elderly Americans, but they are different from the factors based on clinical samples. No age differences in the factor scores were found among the normal elderly subjects when education was considered. In contrast, significant differences were found between a group of 25 demented patients and a matched group randomly extracted from the normal sample. PMID- 10833730 TI - Coaches' expectations and beliefs regarding benefits of youth sport participation. AB - Although many have investigated why children participate in sport, little is known about what adults expect children to gain from participation. The present purpose was to examine coaches' expectations of what children should gain from participation in sport and the extent to which coaches believe that these expectations are actually fulfilled. Participants included 109 youth sport coaches who completed a survey packet consisting of a demographic information questionnaire and the Ohio Sport Satisfaction Index. Analysis indicated coaches ranked the variables of having fun, learning life skills, being part of a team, developing confidence, and the excitement of competition as the most important outcomes for the youth they coach. Generally, coaches believe that their expectations are being fulfilled. Coaches' sex and years of coaching were not significantly related to any of the criterion variables in the present study. PMID- 10833731 TI - Decrement in the horizontal-vertical illusion: are subjects aware of their increased accuracy? AB - 47 subjects adjusted the extended vertical lines of 20 inverted-T figures to make them appear equal to a horizontal line and rated the confidence in their accuracy after each trial. One group viewed figures of varying sizes, a second group viewed figures of standard size, and a third group viewed figures of standard size but received feedback on their accuracy immediately after completing Trial 5. Except for a significant increase in accuracy on Trial 6 for the Feedback Group, there were no differences in performance among groups. Subjects made the vertical line significantly shorter than the horizontal line on initial trials but their accuracy improved over trials. In contrast, there was no consistent increase in confidence, and several analyses indicated that confidence ratings were unrelated to accuracy. These results suggest that the subjects were unaware of the decrement in illusion that occurred over trials. PMID- 10833732 TI - Competitiveness: relations with GPA, locus of control, sex, and athletic status. AB - A sample of college students and college athletes completed measures of competitiveness and locus of control. In addition, self-reports of GPA and sex were provided. Negative correlations among scores on competitiveness, GPA, and scores on internal locus of control were significant. Also, there were sex and athletic status differences in competitiveness. PMID- 10833733 TI - Direction of human motor responses by men and women to aversive stimulation. AB - The frequency of extensions and flexions of the arms of 12 men and 12 women (ages 20-30 years) responding to a neutral tone or to an electric shock was recorded. Subjects had to choose between pushing or pulling a lever upon receipt of an acoustic signal which was paired or unpaired with an electric shock. They were instructed to perform either long duration movements, allowing for on-line control of the execution, or short duration movements with prior specification of amplitude. Regardless of duration of movements, the aversive signal increased the frequency of extensions and intraindividual variability of choices of the men but decreased the frequency of extensions and intraindividual variability of choices of the women. These findings show that stimuli such as pain or fear automatically elicit patterns of terminal motor states corresponding to fight or flight, initiating processes of preparation of spatially oriented movements which are automatic and sex-typed and impair the use of the terminal cues for simultaneous preprogrammed voluntary movements. PMID- 10833734 TI - Preliminary identification of nonlinear correlations for spatial tests and reaction-time tasks. AB - Scores on three spatial tests (mental rotation, embedded figures, and gestalt completion) were significantly correlated with four same-different reaction-time tasks in a sample of 48 female university students. Scores on the Vandenberg-Kuse mental rotation test correlated best with a one-different-all-different task, embedded figures with a one-different-all-same task, and the gestalt completion with a one-same-all-same task, suggesting that the strategies subjects employ for each spatial test are similar to the strategies they use in each of the same different tasks with which their scores correlate best. Present results support the position that stronger correlations than previously noted between scores on spatial tests and reaction-time tasks are observed when matching the tests and tasks in terms of hypothesized underlying processes. PMID- 10833735 TI - Psychosocial stressors and the short life spans of legendary jazz musicians. AB - Mean age at death of 168 legendary jazz musicians and 100 renowned classical musicians were compared to examine whether psychosocial stressors such as severe substance abuse, haphazard working conditions, lack of acceptance of jazz as an art form in the United States, marital and family discord, and a vagabond life style may have contributed to shortened life spans for the jazz musicians. Analysis indicated that the jazz musicians died at an earlier age (57.2 yr.) than the classical musicians (73.3 yr.). PMID- 10833736 TI - Driving-induced stress in urban college students. AB - Urban college student commuters (N = 407) were surveyed about their experiences with stress induced by driving. Of the participants 23.6% reported becoming angry at another driver more than once per day. They rated stress from other drivers as equal to the stress experienced during a college examination but gave slightly lower ratings to traffic congestion, road construction, and finding a parking place as sources of stress. Slow drivers, a child not restrained, and a vehicle following too closely were the highest rated annoying situations. Of participants, 21.6% had reported another driver to the police; nearly 22% said they carried a weapon for protection from other drivers (5.4% said a gun). Men were more than twice as likely as women to carry a weapon and three times as likely to carry a gun. Of the total sample, 19.1% feared being shot by another driver. Most participants (75.8%) said drivers were more aggressive and dangerous than they were five years ago. PMID- 10833737 TI - Behavioral lateralization during spontaneous smelling tasks. AB - The smelling behavior of 52 right-handed subjects was videotaped during tasks involving identification and recognition of different odors. Analysis showed that men more often used the right nostril than the left whatever the odor. There was no significant difference for the women. These results support a more marked cerebral asymmetry in men than in women and a main involvement of the right cerebral hemisphere in the olfactory processes at least by right-handed men. PMID- 10833738 TI - Weather and homicide in Hong Kong. AB - No seasonality was discovered in homicide in Hong Kong. Weather variables were not associated with homicide rates. PMID- 10833739 TI - Attitudes about antidepressants: influence of information about weight-related side effects. AB - Antidepressant drugs are frequently prescribed for women and have various side effects, including potential effects on body weight. This experiment examined the effects of information about the weight-related side effects of antidepressants on women's attitudes toward the drugs. 60 college women were randomly assigned to read about one of two drugs, fluoxetine (Prozac) or imipramine (Tofranil). Participants were either told or not told about veridical weight-related side effects, namely, weight loss for Prozac and weight gain for Tofranil. As hypothesized, weight-gain information lowered the personal acceptability of Tofranil, and weight-loss information enhanced the acceptability of Prozac. Although research with clinical populations is required, undergraduate women's decisions about the use of antidepressant medications may be influenced by societal body-image ideals. PMID- 10833740 TI - Differentiated ratings of perceived exertion and physiological responses during aerobic dance steps by impact/type of arm movement. AB - Overall ratings of perceived exertion, i.e., undifferentiated RPE, are often used as indicators of exercise intensity during walking, jogging, and cycling; however, conflicting results concerning RPE during aerobic dance exercise have been reported, and the use of differentiated RPE, i.e., local RPE and central RPE, has not been investigated. The purposes of this study were to assess local, central, and over-all RPE, and physiological responses [heart rate (HR); % HRmax; absolute and relative VO2;% VO2 max, ventilation (VE), ventilatory equivalent (VE.VO2(-1); and oxygen pulse] during aerobic dance exercise varied by Arm Movement (Static Arm vs Dynamic Arm) and Impact (High vs Low). Trained women (N = 25; max VO2 = 50.4 +/- 7.5 ml.kg-1.min.-1) completed four aerobic dance steps. No RPE were significantly correlated with heart rate or VO2; however, for all steps all RPE were significantly (r = .40-.62) correlated with VE.VO2(-1) or VE. No interactions were present for RPE or physiological variables, and main effects were noted for Impact and Arm Movement. All RPE were greater for High Impact and for Static Arm Movement. Because VE and VE.VO2(-1) were correlated with Overall RPE for all steps, this may suggest that participants "attended to" perceived changes in respiratory phenomena during aerobic dance exercise. It appears that during combined arm-and-leg aerobic dance exercise the use of Overall RPE is sufficient to assess perceptual sensations associated with the intensity of the exercise. Changes in Overall RPE were proportionate to objective measures of exercise intensity, i.e., HR and VO2; however, it is recommended that both HR and Overall RPE be used to assess fully a participant's objective and subjective responses during aerobic dance exercise. PMID- 10833741 TI - Word imageability effects on naming: a pilot investigation of beginning readers of Turkish. AB - Word imageability, a semantic variable, in naming by beginning readers of English is well documented particularly with poor readers naming high imageable words more accurately than low imageable words. The present study examined the role of imageability on word naming by 20 good and 20 poor beginning readers as a function of orthographic transparency by utilizing the peculiarities of the transparent Turkish writing system. Neither good nor poor beginning readers show any evidence of imageability for Turkish suggesting that the contribution of imageability to word naming may indeed be determined by orthographic transparency. Implications of these findings are discussed. PMID- 10833742 TI - Culture and self-presentation as predictors of shyness among Japanese and American female college students. AB - Self-presentation theories of shyness have been supported in North American samples but have not been evaluated cross-culturally. This study examined the relative influence of cultural and psychological variables on self-reported shyness among Japanese and American college students. 35 female Japanese-born and 47 United States-born Euro-American female students completed the Shyness Scale, Rosenberg Self-esteem Scale, Interpersonal Competence Questionnaire, Sensitivity to Rejection Scale, and Individualism-Collectivism Scale, and a demographic data sheet. After statistically controlling for Individualism-Collectivism, psychological measures, especially perceived interpersonal competence and sensitivity to rejection, combined for Adjusted R2 = .32 in shyness. Findings suggest that similar factors are central to experiences of shyness for both samples. Researchers should assess the stability of such findings in larger, heterogeneous samples and evaluate whether treatment strategies that reduce expectations of rejection and increase perceived interpersonal competence have comparable efficacy in reducing shyness across cultures. PMID- 10833743 TI - Lack of sex differences on the Stroop Color-Word Test across three age groups. AB - The present study investigated recent reports of sex differences on the Stroop Color-Word Test by age. Present results indicate no sex differences at 7-8 years, 9-10 years, and 18-24 years. The two school-age samples reported similar amounts of interference and significantly more than the college-age sample. Inconsistencies in the literature may be a function of response modality rather than interference. PMID- 10833744 TI - Perceptions of violent crimes in relation to sex of perpetrator. AB - The purpose of the present study was to examine the relationship between criminal behavior and perceived sex of the perpetrator. Participants (26 men and 44 women) reviewed scenarios depicting an armed robbery and a child's murder. 98% of the respondents attributed the armed robbery to a man, whereas an equal number attributed the child's murder to both a man and a woman. The respondents also ascribed the man's behavior to internal characteristics, whereas the woman's criminal behavior was attributed to external situations or to mental illness. These findings are in accord with current sex stereotypes. PMID- 10833745 TI - Review of the rank transform in designed experiments. AB - Despite a plethora of asymptotic and small samples Monte Carlo studies on the failure of the rank transform in designed experiments, Choi in 1998 published a paper praising the technique. The purpose of this reaction is to (a) examine Choi's literature review on the rank transform, (b) provide some "new" results on the rank transform, and (c) summarize the failures of the rank transform. PMID- 10833746 TI - Dynamic visual acuity and coincidence-anticipation timing by experienced and inexperienced women players of fast pitch softball. AB - This study examined the relationship between dynamic visual acuity and coincidence-anticipation timing in 16 inexperienced and 16 experienced women's fast pitch softball players. Pearson-product correlations indicated a low relationship between dynamic visual acuity and coincidence-anticipation timing. The correlations for dynamic visual acuity and coincidence anticipation between experienced and inexperienced dynamic visual acuity were not significant. A significant difference was found between the mean dynamic visual acuity of the two groups, i.e., experienced players had better dynamic visual acuity than inexperienced players. Analysis of variance of constant errors, variable errors, and absolute errors of coincidence anticipation indicated no significant differences between groups or across the three accuracy scores. The interaction between experience and accuracy was not significant. PMID- 10833747 TI - Development of a Greek version of the Sport Motivation Scale. AB - The aim of the present study was to examine preliminarily the validity of a Greek version of the 1995 Sport Motivation Scale of Pelletier, Fortier, Vallerand, Tuson, Briere, and Blais. For 134 athletes the seven subscales had moderate to good internal consistency (Cronbach coefficients alpha from .64 to .78). Confirmatory factor analysis with nested factor models supported the structural validity of the inventory. Moreover, correlations of scores on the subscales with a measure of task and ego orientation as well as with athletes' self-reported effort in training were in the expected direction. PMID- 10833748 TI - Intensity and direction of competitive state anxiety as interpreted by rugby players and rifle shooters. AB - This study reports the findings of part of an ongoing research program examining sports performers' interpretations of competitive anxiety prior to competition. The notion of 'directional perceptions' has questioned the limited utility of examining only the intensity of competitive anxiety responses as has Jones. The purpose of this study was to examine intensity and direction, i.e., interpretation of intensity as facilitative or debilitative, of anxiety symptoms as a function of two types of sport. The types of sport were explosive (rugby league) versus fine motor skills (target rifle shooting). The sample comprised 50 male rugby league participants and 50 target rifle shooters who completed a modified version of the Competitive State Anxiety Inventory-2 prior to competition. Contingency analysis yielded a significant difference in the number of rugby players who reported somatic anxiety as facilitative and the number of rifle shooters who reported somatic states as debilitative. No such differences were evident for cognitive anxiety. Analysis of variance indicated no differences between the two groups on the intensity of cognitive and somatic anxiety, but the performers competing in rugby league interpreted both states as being more facilitative to performance; the rugby league players also had higher scores on self-confidence than the shooters. These findings provide continuing support for the measurement of directional perceptions of competitive anxiety and highlight the importance of examining individual sports. PMID- 10833749 TI - Block design as a measure of everyday spatial ability: a study of ecological validity. AB - This study investigated the effectiveness of the WAIS-R Block Design subtest to predict everyday spatial ability for 65 university undergraduates (15 men, 50 women) who were administered Block Design, the Standardized Road Map Test of Direction Sense, and the Everyday Spatial Activities Test. In addition, the verbally loaded National Adult Reading Test was administered to assess whether the more visuospatial Block Design subtest was a better predictor of spatial ability. Moderate support was found. When age and sex were accounted for, Block Design accounted for 36% of the variance in performance (r = -.62) on the Road Map Test and 19% of the variance on the performance of the Everyday Spatial Activities Test (r = .42). In contrast, the scores on the National Adult Reading Test did not predict performance on the Road Map Test or Everyday Spatial Abilities Test. This suggests that, with appropriate caution, Block Design could be used as a measure of everyday spatial abilities. PMID- 10833750 TI - An assessment of grade inflation in higher education. AB - Considerable research has been conducted on grade inflation and its pervasiveness. Given the significance of grade inflation on the quality of the educational experience and the reputations of colleges and universities, efforts to assess its presence and underlying causes should be supported and solutions developed. Because periodic changes in average grades in the short term may be anomalous, mean grade point averages (GPAs) for 262 undergraduate courses at a Liberal Arts college were examined for trends across a 10-yr. period. Analysis showed higher grades appeared within two of the colleges' four academic units, although the reasons remained unclear. Tentative explanations are explored. PMID- 10833751 TI - An investigation of finger and manual dexterity. AB - In ergonomics, handgrip has been perceived as one of the most important hand functions; however, other types of functions of the hand are also important to the ergonomist, such as finger and manual dexterity, on which few ergonomic studies have been done. This experiment involved 60 male and female college students whose performance (number of pegs placed into a board in 30 sec.) of finger dexterity were statistically significantly different between the right and left hands and between women and men. The interaction of hand and sex was significant. PMID- 10833752 TI - Expertise in women's gymnastic judging: an observational approach. AB - Observation was used to study expertise in gymnastic judging. 10 expert and 10 novice gymnastic judges were videotaped while judging at actual competitions. Analyses showed that novice judges, as compared to expert judges, spent less time looking at the gymnast perform, spent more time looking at the scoring paper, and were less able to engage in the dual-task demands required in gymnastic judging. PMID- 10833753 TI - State anxiety, performance accuracy, and work pace in a simulated pharmacy dispensing task. AB - This study examined the effects of scores on state anxiety on overall accuracy of performance and speed of working in a simulated pharmacy dispensing task. 75 undergraduates worked in a simulated pharmacy environment, designed by Schell and Grasha in 1998, to fill 42 mock orders for simulated pharmacy items. Participants' accuracy and work pace in the simulation, presimulation stress, and postsimulation perceived workload and state anxiety were measured. Analysis indicated that state anxiety and overall accuracy were strongly related. State anxiety appears to be one of the best predictors of errors in the simulated pharmacy dispensing task found so far, while the lack of relationship between work pace and accuracy was confirmed. Work pace predicted accuracy, indirectly, but only after statistically removing the effects of anxiety, task frustration, significant-other stress, and grade point average. PMID- 10833754 TI - Towers of Hanoi and London: is the nonshared variance due to differences in task administration? AB - Although it has been assumed that the Tower of Hanoi and Tower of London are more or less interchangeable tasks dependent on executive function, a series of studies in our laboratory have indicated substantial nonshared variance between the performances on the two tasks. The purpose of the present study was to examine how much methods of administration, such as number of trials per problem, contribute to this nonshared variance. A new one-trial version of the Tower of Hanoi was developed to be identical to the Tower of London in four procedural characteristics. The one-trial version of the Tower of Hanoi was administered to 39 normal adults along with the traditional Tower of Hanoi and the Tower of London-Revised in two test sessions 5-7 weeks apart. The correlations between the two tasks were in the same range as found previously with the traditional task, indicating that administration differences do not account for the nonshared variance between the tasks. A reliability analysis of the one-trial tasks showed poor internal consistency. Also, the internal consistency of the 6-trial tower was artificially inflated by aspects of the administration and scoring procedures. Moreover, this task exhibited a ceiling effect on repeated testing. These results suggest that it would be of value to redesign the one-trial Tower of Hanoi systematically to increase its reliability and, potentially, its validity as a measure of executive functions. PMID- 10833755 TI - Social class difference in response to Christmas cards. AB - In a total of 590 Christmas cards sent perception of status was important for both the sender and the receiver. High status of the sender increased the response rate significantly, especially among the "blue-collar" receivers. PMID- 10833756 TI - Another look at children's symbol reversals. AB - In a previously reported longitudinal study of reversal errors for static and kinetic written symbols we found no compelling support for their academic importance in kindergarten (n = 201), Grade 1 (n = 156), or Grade 2 (n = 129); however, for Grade 3 (n = 105), kinetic reversals became a significant predictor of tested reading achievement. If reliable, this finding might have implications for the identification of children with long-term reading impairment. PMID- 10833757 TI - The biological week and broader time structures (chronomes): in memory of Gunther Hildebrandt. AB - Data illustrating the biological week and chronomes are presented in recognition of extensive work of G. Hildebrandt. PMID- 10833758 TI - Environmental influences on perceptual and motor skills of children from immigrant Ethiopian families. AB - The purpose of this study was to examine environmental influences on perceptual and motor skills of children from immigrant Ethiopian families in Israel in comparison with those of Israeli-born children. The subjects were divided into groups based on age (6- to 8-yr.-olds and 10- to 12-yr.-olds) and length of time in Israel. The results of this study show that the perceptual and motor performance of 6- to 8-yr.-old, less recent Ethiopian immigrants is similar to that of the recent Ethiopian immigrants of the same age. A difference exists between those two groups and the Israeli-born children of the same age. In the 10 to 12-yr.-old age group, there is a marked difference between the performance of the two groups of Ethiopian immigrants. The difference between the less recent immigrants and the Israeli-born children is smaller for most tasks; however, the less recent immigrants' performance is still poorer than that of the Israeli children. These results are compatible with those of other studies describing the difficulties encountered by children who had not studied in formal educational settings and those who have had that experience. PMID- 10833759 TI - Petter's effect in patterns formed by outlined surfaces. AB - In patterns formed by two equally colored or by two transparent overlapping surfaces of different size that alternately appear in front of one another, the larger surface has a greater probability of appearing in front of the smaller surface. This effect is known as Petter's effect. The present study found that Petter's effect also occurred in patterns formed by colorless outlined surfaces. In these patterns Petter's effect was smaller than in chromatically homogeneous patterns. The results agree with the possibility that Petter's effect occurred in patterns formed by outlined surfaces because relative size was a cue to visually perceived distance. PMID- 10833760 TI - The Groningen Enjoyment Questionnaire: a measure of enjoyment in leisure-time physical activity. AB - The purpose of this study was to develop a short and easily used questionnaire to measure enjoyment in leisure-time physical activity. The first part of the study involved the questionnaire's composition. A set of 30 positively formulated potential items was generated and subsequently completed by 59 subjects. We used these results to trim the questionnaire to 10 items. In the second part of the study, the questionnaire was administered to 82 subjects and research was done into reliability and validity. The results indicate that the Groningen Enjoyment Questionnaire can be considered reliable and valid. PMID- 10833761 TI - Comparison of selected perceptual variables for backpacks with internal and external frames. AB - Differences in perceptions of comfort, exertion, balance, and heart rate were investigated with two types of backpack. Subjects were 20 male volunteers (M age = 24.3 yr., SD: 3.6, M height = 180.02 cm, SD: 8.0, and M weight = 86.46 kg, SD: 14.84). Following oral briefing and practice trials, each subject was fitted at random with either an internal- or external-frame backpack containing 18.2 kg. By random, cross-over design, subjects completed a 30-m simulated hiking trial consisting of 16 separate obstacles designed to resemble actual off-trail hiking. Trials were conducted in a controlled environment with the walking intensity regulated by a metronome. Upon completion of each trial, the subjects rated the experience on perceived comfort, balance and stability, and rating of perceived exertion. Heart rate was recorded immediately prior to and following each trial. Analysis yielded a significant difference only on rating of perceived exertion in favor of the internal-frame backpack. Under limited circumstances, these data suggest that both types of backpacks provide similar comfort and balance and stability; however, internal frames in backpacks may reduce perceived exertion during short, demanding hikes. PMID- 10833762 TI - Aggression: destructive and constructive aspects. AB - The concept of aggression was long relegated to the shadow of the libido in psychoanalytic theory, placed among the negative affects by some theoreticians, denied the role of a primary drive among leading writers in clinical psychology, brought to the fore as an adaptive force in ethology, and proved to be a decisive factor in the development of psychosomatic ailments. The second part of the paper reviews experiments using projective techniques, showing that highly creative subjects as opposed to ones low in creativity seem inclined to accept their aggressive impulses. A total denial of these impulses has been typical of women with breast cancer. PMID- 10833763 TI - Effects of smoking abstinence on movement regulation. AB - The effect of smoking abstinence on performance of a reciprocal tapping task was investigated. 6 habitual smokers performed a single-plate and two versions of a two-plate tapping task. Fitts' Law was used to compute an index of difficulty (ID) in bits for the tasks which was 0 bits for the single-plate and 3.32 and 4.17 bits for the two-plate versions of the task. While smoking abstinence had no effect on performance of the single-plate tapping task, it increased movement time on performance of both two-plate task versions. These findings may provide a coherent explanation for the prior findings of nicotine deprivation on psychomotor performance in the literature. This explanation suggests that the effects of nicotine deprivation as incurred through smoking abstinence may be on the central mechanisms regulating information-processing rate for successful movement regulation. Thus nicotine deprivation may not affect performance of simple psychomotor tasks which require minimal information processing but will affect the performance of more complex tasks requiring significantly more information processing for successful movement regulation. PMID- 10833764 TI - Auditory processing and cluttering in young children. AB - This study examined the differences among scores on four tests of auditory processing of 6 children who clutter and 6 control subjects matched for age, sex, and grade. Scores on a consonant-vowel dichotic listening task indicated that directing the attention of the attended ear improved the percentage of correct responses for both groups of children. Those who clutter, however, showed a greater percentage of change during the directed right and left ear conditions. Cluttering children performed poorer on right and left competing conditions of the Staggered Spondaic Word Test. No differences were found between groups for the auditory attention task and the time-compressed speech task. Implications for processing of dichotic stimuli and diagnosis of children who clutter are discussed. PMID- 10833765 TI - Is defense or offense more important for professional football teams? A replication study using data from the 1998-1999 regular football season. AB - The purpose of the present investigation was to replicate Onwuegbuzie's 1999 study by identifying which variables best predict a team's winning percentage in the 1998-1999 regular professional football season. Findings gave a model containing five variables: (1) turnover differential (which explained 54.4% of the variance), (2) total number of rushing yards conceded by the defense (which explained 21.3% of the variance), (3) total number of passing first downs attained by the offense (which explained 9.4% of the variance), (4) percentage of third-down plays that produce a first down (which explained 4.1% of the variance), and (5) total number of penalties conceded by the opponents' defense resulting in a first down (which explained 4.1% of the variance). These variables combined to explain 93.3% of the total variance in winning percentage for the 1998-1999 season. That the first two variables, which explained more than 75% of the variance, were characteristics of the defense supports Onwuegbuzie's conclusion that defensive gains are better predictors of success than are offensive gains. PMID- 10833766 TI - Professional cricketers' perceptions of the importance of antecedents influencing repeatable good performance. AB - This study examined whether there is overall agreement amongst professional cricketers on the factors which are most important for the occurrence of repeatable good performances in the sport. Also, the consistency of the rank order of such variables was investigated across individuals and roles within the same sport. A total of 198 cricketers (100 batters and 98 bowlers) who play or have played first class cricket in the English County Championship completed a rank-order task, which enabled them to use their experiences to judge the importance of 15 selected performance-influencing variables. Cultural consensus analysis indicated that no one factor for either batters or bowlers was significantly more important, despite many batters and bowlers ranking the need for 'total self-confidence' highest. Whilst no over-all agreement was found for batters and bowlers in their ranking of variables, a group of similar 'core elements', e.g., total self-confidence, following a set prematch routine, set performance plans, optimal arousal, and using mental rehearsal, were highly ranked by both groups, which may be an important consideration for practitioners endeavoring to produce strategies that encourage more consistent and higher performance for cricketers. PMID- 10833767 TI - Experimental simulation of a haunt experience and elicitation of paroxysmal electroencephalographic activity by transcerebral complex magnetic fields: induction of a synthetic "ghost"? AB - To test the hypothesis that experiences of apparitional phenomena with accompanying fear can be simulated within the laboratory, a 45-yr.-old journalist and professional musician who had experienced a classic haunt four years previously was exposed to 1 microTesla, complex, transcerebral magnetic fields. Within 10 min. after exposure to a frequency-modulated pattern applied over the right hemisphere, the man reported "rushes of fear" that culminated in the experience of an apparition. Concurrent electroencephalographic measurements showed conspicuous 1-sec.-to-2-sec. paroxysmal complex spikes (15 Hz) that accompanied the reports of fear. A second magnetic field pattern, applied bilaterally through the brain, was associated with pleasant experiences. The subject concluded that the synthetic experience of the apparition was very similar to the one experienced in the natural setting. The results of this experiment suggest that controlled simulation of these pervasive phenomena within the laboratory is possible and that this experimental protocol may help discern the physical stimuli that evoke their occurrence in nature. PMID- 10833768 TI - Effects of learning and movement frequency on polyrhythmic tapping performance. AB - This study examined the effect of learning a complex bimanual coordination task at different movement frequencies. 30 subjects performed 5:3 polyrhythmic tapping at either high, medium, or low movement frequency on a rhythmic synchronization task and then reproduced the polyrhythmic pattern repeatedly in the spontaneous task. Analysis showed that practice on the synchronization task qualitatively changed correct responses into anticipatory ones. The synchronization learning of the polyrhythm caused the anticipatory responses and so, may involve memorization of serial positions within the polyrhythm. Also, more anticipatory responses were indicated in performance at the medium and low frequencies than at the high frequency on the synchronization task. In addition, deviations of taps from expected tapping positions were observed in performance of the spontaneous task at the high frequency. These results suggest that the movement frequency qualitatively influenced the learning of this bimanual coordination. Especially at the high frequency, frequent shifts to other coordination patterns occurred on the spontaneous task. This means that the performance at higher frequency is more strongly affected by entrainment between the two hands. PMID- 10833769 TI - Relationship between emotional state and performance during international field hockey matches. AB - The present study examined the relationship between the emotions experienced by 15 international hockey players, both immediately before and during competition, and their performance levels. Data were collected on the players' emotional states using a revised version of the Feelings Scale of Butler, which was completed retrospectively after the match was played. Players reported more annoyance and less tension during the match than before. A logistic regression correctly classified 70.2% of players from the emotional ratings immediately before the match and 85.1% of the players from the ratings during the match as either a good or poor performer. Those individuals who performed well retrospectively reported feeling Nervous and 'Quick/Alert/Active' before the game and Confident and Relaxed during the game. The results indicate that emotions fluctuate over the competition period, and in long duration sports assessment of emotion during competition predicts variation in performance better than assessment prior to competition. PMID- 10833770 TI - The Zeigarnik effect in a sample of mentally retarded persons. AB - A conceptual replication of the well known Zeigarnik effect was attempted with persons who had organic brain dysfunction. Data from a sample of 56 mentally retarded persons showed a significant difference in the recall of uncompleted versus completed tasks, thus establishing the Zeigarnik effect in that population. PMID- 10833771 TI - C-reactive protein: a story half told. PMID- 10833772 TI - Quantitating urea removal in patients with acute renal failure: lost art or forgotten science? AB - A 67-year-old woman is admitted to the surgical service with a high fever, a painful and distended abdomen, jaundice, and almost complete anuria. A urinalysis revealed dark red-brown urine notable for albuminuria, erythrocytes, leukocytes, and casts. The patient was treated with antibiotics, but continued to have oligoanuria. On the eighth day of hospitalization, the following laboratory tests were obtained: serum potassium, 13.7 mEq/L; BUN, 396 mg/dl. At this time the patient was noted to be encephalopathic with deteriorating clinical condition. Renal replacement therapy was initiated. The characteristics of the initial dialysis treatment are described in Table 1. After the initial dialysis treatment, the patient went on to become nonoliguric, followed by gradual recovery of urea clearance. She survived her acute illness, left the hospital, and at 7 months posthospitalization was doing quite well. PMID- 10833773 TI - The overlooked role of salt restriction in dialysis patients. PMID- 10833774 TI - The role of the bone biopsy in the diagnosis of renal osteodystrophy. PMID- 10833775 TI - Avoiding the use of hypertonic dextrose dialysate in peritoneal dialysis. PMID- 10833776 TI - The problem of compliance with PD exchanges. PMID- 10833778 TI - Management of uremic pruritus. PMID- 10833777 TI - What are the causes and consequences of the chronic inflammatory state in chronic dialysis patients? PMID- 10833779 TI - Anticoagulation for intermittent hemodialysis. PMID- 10833780 TI - Pathogenic effects of a high peritoneal transport rate. AB - Recent studies have shown that patients with high peritoneal transport characteristics have substantially increased morbidity and mortality. This finding is counter-intuitive, since HTs will a priori achieve higher clearances. There are many possible causes: increased protein losses with consequent hypoalbuminemia; poor ultrafiltration capacity causing fluid retention, ventricular hypertrophy and hypertension; increased glucose absorption leading to anorexia, hyperinsulinism, and local AGE formation; and the development of an atherogenic lipid profile. While common pathogenic causes of high peritoneal transport and atherosclerosis have been hypothesized, it is more likely that CAPD as currently practiced is unsuitable for HTs, who should be switched to HD or NIPD. Renal and peritoneal clearances have different clinical effects and should be assessed separately. Current measures of dialysis adequacy, such as total Kt/V, do not therefore describe the patient's clinical situation accurately and are insufficient. PMID- 10833781 TI - Recurrent and de novo diseases after renal transplantation. PMID- 10833782 TI - Donating a second kidney: a tale of family and ethics. PMID- 10833783 TI - BP control, dialysis adequacy, and vitamin C. PMID- 10833784 TI - Failure of central vein catheters after prolonged use. PMID- 10833785 TI - Reducing infections associated with central vein catheters. PMID- 10833786 TI - Use of intraperitoneal insulin with CCPD. PMID- 10833787 TI - Pneumococcal vaccine with unknown vaccination status. PMID- 10833788 TI - [Arterial hypertension or essential hypertension: arguments for nosological diagnosis]. PMID- 10833789 TI - [Application of DNA analysis for differential diagnosis of familial hypercholesterolemia and familial defect of apolipoprotein b-100]. AB - AIM: To determine occurrence of apolipoprotein B-100 mutation in codon 3500 (apoB3500) in patients with primary hypercholesterolemia in Russia. MATERIALS AND METHODS: The study included 71 patients with clinical diagnosis of familial hypercholesterolemia (FH) and 24 relatives. All the subjects were tested for the presence of apoB3500 mutation using polymerase chain reaction and cleavage with restriction enzyme HhaII. Samples demonstrating anomalous pattern were further analysed by automatic DNA sequencing. RESULTS: Apob3500 mutation was detected in two (2.8%) female patients. In both cases cholesterol levels were severely increased although clinical features were different. CONCLUSION: Some cases of primary hypercholesterolemia in Russia may be due to familial defective apoB-100. Further screening of FH patients is required for a precise estimation of the incidence rate of familial defective apoB-100 in this country. PMID- 10833790 TI - [Determination of coronary-myocardial reserve by stress echocardiography in patients with programmed pacemaker]. AB - AIM: To study coronary-myocardial reserve in patients with a programmed pacemaker. MATERIALS AND METHODS: Stress-echocardiography was performed in 64 patients with coronary heart disease (CHD). The stress was induced by a gradual increase in the frequency of electrostimulation (ES) by 10 imp/min from initial 90 imp/min to threshold value which was defined as the frequency threshold of myocardial ischemia induction (FT). RESULTS: Registration of defects in local left ventricular contractility and cardiodynamics in frequent heart ES identified patients with predominant coronary and myocardial failure. Low FT (100-110 imp/min) indicates poor coronary reserve while a considerable rise of the end diastolic pressure in the left ventricle indicates limited myocardial reserve. Positive results of the test in isolated ventricular ES were obtained in 90.9%, in atrial ES in 72.7% of patients. CONCLUSION: As a highly informative and reproducible method, stress echocardiography can be employed for optimization of antianginal therapy in CHD patients with a pacemaker. PMID- 10833791 TI - [Soluble P-selectin - a marker of platelet activation and vessel wall injury: increase of soluble P-selectin in plasma of patients with myocardial infarction, massive atherosclerosis and primary pulmonary hypertension]. AB - AIM: A comparative analysis of the content of the soluble form of cell adhesion protein P-selectin in the blood plasma of patients with acute myocardial infarction (AMI), massive atherosclerosis (MA) and primary pulmonary hypertension (PPH), investigation of the relationship between plasma content of P-selectin and known markers of platelets and endothelial cells activation, preliminary assessment of the prognostic value of P-selectin determination. MATERIALS AND METHODS: This study included 16 patients with AMI, 20 patients with MA, 21 patients with PPH and 18 healthy donors. The follow-up was 1-5 years. End-points in the group of patients with AMI were recurrent acute coronary syndrome and coronary artery by-pass operation, in the group with MA--thrombotic complications (acute coronary syndrome, ischemic stroke) and in the group with PPH--death. P selectin was measured by ELISA and platelet factor 4 (PF4), thromboxane B2 (TXB2), endothelin-I and stable prostacyclin metabolite 6-keto-prostaglandin F1 alpha (6-keto-PGF1 alpha) by means of commercial ELISA kits. RESULTS: Mean level of P-selectin in blood plasma of patients with AMI (1 day) (361 +/- 18 ng/ml), MA (410 +/- 31 ng/ml) and PPH (627 +/- 83 ng/ml) was increased in comparison with the group of healthy donors (269 +/- 12 ng/ml) (everywhere p < 0.001). In AMI, P selectin was increased on day 1 only, on days 2, 3 and 10-14 of the disease the level of P-selectin was significantly lower than on day 1 and did not differ from the control level in the group of donors. In patients with MA a significant correlation was detected between plasma content of P-selectin and platelet activation marker PF4 (r = 0.606, P = 0.007) and in patients with PPH between the content of P-selectin and another platelet activation marker TXB2 (r = 0.622, p = 0.013). However, no correlation was found in PPH patients between the content of P-selectin and markers of endothelial activation and/or damage (endothelin-1 and 6-keto-PGF1 alpha). Difference in the concentration of P-selectin in patients with or without end-points during the follow-up period was detected in patients with AMI (353 +/- 14 ng/ml and 451 +/- 24 ng/ml, p = 0.009) and PPH (477 +/- 58 ng/ml and 927 +/- 184 ng/ml, p = 0.017) but not with MA (426 +/- 37 ng/ml and 361 +/- 24 ng/ml, p = 0.295). CONCLUSION: The level of P-selectin in plasma was increased in patients with acute thrombosis (AMI, 1 day) as well as in patients without clinical signs of thrombosis but with a massive injury of the vasculature (MA and PPH). The increase of P-selectin was, presumably, caused by its secretion from activated platelets since its concentration in plasma correlated with platelet concentration but not endothelial activation markers. Preliminary data indicate that blood plasma soluble P-selectin may be considered as a potential prognostic marker in AMI and PPH. PMID- 10833792 TI - [Purine metabolism enzymes in diagnosis and differential diagnosis of osteoarthritis and gout arthritis]. AB - AIM: To characterize purine metabolism in osteoarthritis (OA) and gout arthritis (GA) diagnosis and differential diagnosis of these diseases. MATERIALS AND METHODS: We estimated xanthine oxidase (XA), xanthine dehydrogenase (XDG), 5' nucleotidase (5'-NT) activity, esoenzymes of XDG and content of uric acid (UA) in the sera of 44 patients with osteoarthritis and 34 patients with gout arthritis. RESULTS: Hyperuricemia was revealed in 25 percent of patients with osteoarthritis, in 64.7 percent of patients with gout arthritis. XO, XDG, 5'NT activity, XO/XDG activities ratio, XDG-2 esoenzymes and content of UA were increased in OA patients compared to healthy controls. XO, XDG activity, XDG-2 esoenzymes and UA content in GA patients were higher than those in OA patients. CONCLUSION: The enzyme difference found may promote differential diagnosis of OA and GA. The enzyme indices essentially depend on clinical specificity of the disease and can be helpful in the assessment of the treatment efficiency. PMID- 10833793 TI - [Diagnosis of Behcet disease: clinical examples]. PMID- 10833794 TI - [Polymorphism of vascular angiotensin II receptor gene and cardiovascular disorders]. AB - AIM: To study polymorphism of the gene of vascular angiotensin II receptor. MATERIALS AND METHODS: Polymorphism that consists in variability of adenine (A) and cytosine (C) residues at position 1166 of the gene for vascular angiotensin II receptor (AT1R) was analyzed in a Moscow population (n = 98) and three groups of affected patients with myocardial infarction (n = 32, MI), left ventricular hypertrophy (LVH, n = 38) and essential hypertension (EH, n = 178). Polymorphic region of the AT1R gene was amplified using the polymerase chain reaction (PCR) and genomic DNAs from human whole blood as template. PCR products were electrophoresied in a gel after digestion with BstDEI restriction nuclease. Significance of differences in distribution of both allele and genotype frequencies at the population sample and in affected patients were estimated via exact Fisher's test. RESULTS: A significant decrease in the frequency of the A genotype was detected in all the three affected groups compared to healthy controls. Besides, the frequency of the A allele was significantly decreased in EH group with a corresponding increase in the frequency of both the AC genotype and the C allele. CONCLUSION: The A1166C polymorphism of the AT1R gene is associated with EH, MI and LVH in a Moscow population. The association is stronger with EH. The A allele and the AA genotype protect against development of disorders at early onset while the other genotypes and the C-allele are risk factors. A protective role of the AA genotype is more significant than predisposition action of the CC homozygote. PMID- 10833795 TI - [Vegetovascular dysfunction as an integral parameter of high risk for cardiovascular diseases development in young subjects]. AB - AIM: To study manifestations of vegetative dystonia in children of probands from families with hereditary IHD load regarding the main risk factors. MATERIALS AND METHODS: 44 individuals aged 21 +/- 0.7 years were selected out of 111 children from 103 examined families the fathers of which had transmural or macrofocal myocardial infarction at the age under 50. 22 of 44 examinees had syndrome of vegetovascular dystonia (VVD). RESULTS: Children of probands with family history of cardiovascular disease and having VVD had levels of cholesterol, triglycerides and apoB higher, but HDLP cholesterol lower than those free of VVD. In daughters of the probands insulin and hydrocortisone levels were high whereas sons with VVD had only insulin levels higher. CONCLUSION: Children of probands of both sexes had high coefficients of hormonal and metabolic adaptation dependent on the severity of VVD syndrome. PMID- 10833796 TI - [Results of 24-hours arterial blood pressure monitoring in patients with mitral prolapse and anomalous chorda]. AB - AIM: To examine 24-h arterial hypertension (AH) profile in patients with connective tissue dyplasia syndrome (CTDS). MATERIALS AND METHODS: 92 patients (35 males and 57 females) with CTDS aged 18-37 years. 68 had mitral prolapse stage I-II, of them 23 had anomalous chorda. 25 patients with vegetovascular dystonia (VVD) free of cardiac lesions. 24-h monitoring of arterial pressure was done in all the patients. RESULTS: Hypertensive load was higher in CTDS and VVD being the highest in patients with the prolapse. Incidence of arterial hypertension was similar in all the groups. CONCLUSION: CTDS patients have indications for 24-h monitoring of arterial pressure to reveal latent arterial hypertension and profile of arterial pressure. This will allow to predict complications of hypertension and prescribe relevant treatment. PMID- 10833797 TI - [Population study of ischemic heart disease socio-psychological risk factors in male population of Novosibirsk]. AB - AIM: To study a 5-year prevalence of and changes in anxiety, behavior of the coronary type, social support, health self-estimation, attitude to smoking, sleep disorders in male population. MATERIALS AND METHODS: In the context of the program "MONICA-psychosocial", a random representative sample of males aged 25-64 living in Novosibirsk has been examined (screening in 1988-1989--739 males, and in 1994-1995--657 males). RESULTS: Personal anxiety in the study population has increased for 5 years from 49.5 to 51%. The number of males with coronary behavior has reduced from 25.9 to 16.9% (p < 0.001). Health self-estimation has improved and frequency of sleep disorders diminished. Negative tendencies also manifested: less number of males quit smoking, social connections and contacts decreased. CONCLUSION: A close correlation between psychosocial factors and ischemic heart disease necessitate intensification of the preventive programs aimed at weakening of influence of psychosocial factors on the population. Otherwise no reduction in ischemic heart disease incidence in population can be expected. PMID- 10833798 TI - [Data of one-stage screening trial of an open Tumen population]. AB - AIM: To evaluate epidemiological situation by cardiovascular diseases and their risk factors in Tyumen. MATERIALS AND METHODS: An open population of the Central Tyumen district was studied (1608 patients) for prevalence of obesity, arterial hypertension, systolic and diastolic arterial pressure. RESULTS: It was found: high incidence of arterial hypertension both in males and females (25.6 and 24.6%, respectively) with predominance of isolated systolic arterial hypertension in younger patients, high incidence of borderline hypertension in young men, higher predisposition to obesity in males vs females. Body mass index was higher in 25-34-year-old males but increased with age in females. CONCLUSION: It is necessary to perform a cohort study of epidemiological parameters to detect persons with cardiovascular disease risk. PMID- 10833799 TI - [Morning rise of systolic blood pressure (by 24-hour ambulatory monitoring) and platelet aggregability in essential hypertension patients]. AB - AIM: To investigate the relationship between platelet aggregability (PA) and parameters of blood pressure (BP) in patients with essential hypertension (EH). MATERIALS AND METHODS: We analyzed 24-h BP recordings (SL-90207, 15-min day and 30 min night time intervals) of 47 hospitalized males with mild to moderate EH (mean age 48 +/- 1 years) to assess the following parameters: mean 24-h, awake (Aw) and nighttime (N) systolic (S) and diastolic (D) BP. We assessed the morning rise (MR) of BP using the new index: a relative morning rise of systolic BP RMRSBP--(max value of SBP from 6 am to 12 am/mean asleep SBP) x 100%. The kinetics of mean aggregate size (MAS) changes was studied with aggregation analyzer model (230LA Biola Ltd., Russia). The following parameters were used for estimation of platelet aggregability: a relative increase in MAS 2 min after beginning of sample stirring--for spontaneous aggregation (SPA) and the maximum increase in the light transmission for 0.5 microM ADP-induced aggregation (ADPI PA). The patients were divided into two groups according to the median value of RMRSBP: group 1 (n = 25, RMRSBP < 121%) and group 2 (n = 22, RMRSBP > 121%). The differences in estimated parameters were tested by Student two tailed t-tests and presented by M +/- SE. P < 0.05 was considered statistically significant. RESULTS: No significant differences have been found between the groups by mean age, body mass index, duration of arterial hypertension, mean 24-h, awake DBP and SBP. Statistically significant differences have been found between groups by SPA, ADPI-PA, night SBP, night DBP, RMRSBP, RMRDBP. In group 2 there was a correlation between RMRSBP and SPA, but not in group 1. CONCLUSIONS: The morning rise of systolic BP is associated with an increase of ADP-induced and spontaneous platelet aggregability in the patients with mild to moderate essential hypertension and apparently that association is more pronounced at high values of morning BP (more than 20% from mean nocturnal values of SBP). PMID- 10833800 TI - [Activity of intracellular antioxidant enzymes in hypertensive patients]. AB - AIM: To assess activity of oxidation and function of cell antioxidant system in hypertensive patients. MATERIALS AND METHODS: 24 hypertensive patients (AP 140 180/95-114 mm Hg) were examined before and 4 weeks after standard therapy for plasmic levels of thiobarbituric acid (TBA) reacting products (TBARP), lipid peroxides (LP), total oxidant activity (TOA), total antioxidant activity (TAA) in red cells, activity of superoxide dismutase and catalase. A control group consisted of 16 healthy volunteers matched for age and sex. RESULTS: The patients had very high TAA and TBARP while the antioxidant status was subnormal. TOA/TAA was higher than in the controls. Clinically effective therapy with basic antihypertensive drugs lowered the oxidant activity and raised the antioxidant status while TOA/TAA index remained elevated. CONCLUSION: Hypertensive patients are exposed to apparent oxidant stress caused by imbalance between adaptive potential of intracellular enzyme antioxidant defense and activity of free radical processes. It is suggested that in pathogenesis of essential hypertension insufficiency of antioxidant enzymes results in development of chronic oxidant stress. PMID- 10833801 TI - [Clinical manifestation of hypertension in citizens of siege Leningrad (1941 1944). Retrospective analysis of archive materials]. AB - AIM: To characterize peculiarities of arterial hypertension course in citizens of sieged Leningrad. MATERIALS AND METHODS: 2000 case records of hypertensive patients treated in 6 hospitals of sieged Leningrad have been analysed. RESULTS: Arterial hypertension (AH) was verified as the basic disease in 69 cases. Of them, only 47 patients were eligible for analysis. Mean age of the patients was 45 years. AH duration before hospitalization was less than 1 year in 35% of the cases. Hypertensive crises, hypertensive angiopathy, cerebral atherosclerosis, cardiac hypertrophy were documented in 25, 15, 15 and 64% of the cases, respectively. In hospital, no specific antihypertensive therapy was given. At the discharge, systolic blood pressure decreased significantly, diastolic blood pressure decreased insignificantly. CONCLUSION: Clinical data evidence for rapid affection of target organs in hypertensive subjects exposed to unfavourable conditions of the war time. PMID- 10833802 TI - [Course of pulmonary hypertension in patients operated for rheumatic mitral heart disease]. AB - AIM: To study changes in pulmonary hypertension in patients with isolated mitral valvular defects of rheumatic etiology after surgical correction and in the early postoperative period. MATERIALS AND METHODS: Central hemodynamics in heart chamber probing before operation and intraoperatively were measured in 98 patients with rheumatic mitral defects (41 males, 57 females, mean age 39.5 +/- 8.74). Manometry before and after the defect correction, intraoperative catheterization (cath. Swan-Ganz) for hemodynamics 24-h follow-up were made. RESULTS: After valvular defect correction 90(91%) patients had residual pulmonary hypertension, stage II-III in 41%. CONCLUSION: Patients operated for rheumatic heart disease complicated by pulmonary hypertension often suffer from residual pulmonary hypertension. This requires pharmacological correction. PMID- 10833803 TI - [Acetylsalicylic acid effects on pain sensitivity to myocardial ischemia and skin sensitivity in patients with angina pectoris]. AB - AIM: To clarify if pain-relieving action of acetylsalicylic acid (ASA) is associated with lowered sensitivity of anginal patients to pain due to myocardial ischemia. MATERIALS AND METHODS: A double blind randomized placebo-controlled trial enrolled 10 males aged 42-69 years with stable effort angina (EA) of functional class II-III. When exposed to exercise tolerance test (treadmill, stress-system Sicard 460S, computed ECG), the patients developed EA attack with at least 1 mm decline of ST segment on ECG. The exercise test was made before, 2 and 4 hours after administration of ASA and placebo. Sensitivity to ischemia was estimated by the total depth of the ST segment decline in 11 ECG leads (sigma ST) registered at the attack onset. Tactile and pain thresholds (TT and PT) were studied with a highly reproducible technique. TT and PT were measured before, 2 and 4 hours after ASA and placebo administration. RESULTS: 2 and 4 hours after intake of 100 mg of ASA, sigma ST and TT significantly rose compared to the baseline level and placebo. PT significantly rose vs the baseline level. CONCLUSION: ASA deteriorates sensitivity of anginal patients to myocardial ischemia, skin tactile and pain sensitivity and thus can deprive the EA patient of the pain attack signal. This leads to the risk of overexercising and emergence of painless myocardial ischemia. PMID- 10833804 TI - [New data on etiology of heart conduction disturbances]. AB - AIM: To make a clinicogenealogical analysis of different types of heart conduction disturbances in families of Krasnoyarsk city. MATERIALS AND METHODS: 104 probands with ECG-verified idiopathic forms of atrioventricular and intraventricular blocks of different severity and 331 their close relatives (kinship degree I and II) were examined using standard clinical investigations, ECG, echocardioscopy, electrophysical investigations in some cases and coronarography. RESULTS: All the observed 104 families were divided into 3 groups according to the type of heart conduction in proband. Group 1 consisted of 24 probands with atrioventricular block and 80 their relatives. The sick relatives had for the most part atrioventricular blocks (31.2%). 26 probands of group 2 had complete right bundle branch blocks. Of their 81 relatives, the sick ones had primarily conduction disturbances in right bundle branch (incomplete block- 44.7%, a complete block--5.2%). Group 3 consisted of 54 probands with left bundle branch block and 170 relatives. The left bundle branch block was present in 27.4% of the relatives. CONCLUSION: A definite family aggregation of heart conduction disturbances was found. This proves genetic determination of this pathology. PMID- 10833805 TI - [ACE inhibitors in cardiac failure: from research to routine practice. (lecture)]. PMID- 10833806 TI - [Arterial hypertension and quality of life: role of life quality assessment in clinical studies and practical medical activities]. PMID- 10833807 TI - [Genetic aspects of chronic cardiac failure]. PMID- 10833808 TI - [Immuno-inflammatory activation as a conceptual model of development and progression of cardiac failure]. PMID- 10833809 TI - Immigration, HIV infection, and sexually transmitted diseases in Europe. PMID- 10833810 TI - HIV-associated psoriasis. AB - Psoriasis occurs with at least undiminished frequency in HIV-infected individuals. The behavior of psoriasis in HIV disease is of interest, in terms of pathogenesis and therapy because of the background of profound immunodysregulation. It is paradoxical that, while drugs that target T lymphocytes are effective in psoriasis, the condition should be exacerbated by HIV infection. The etiopathogenesis of psoriasis is unknown, but genetic and environmental factors are thought to be involved. There are controversial issues regarding the immunological basis of psoriasis and the role of CD4+ versus CD8+ T lymphocytes. Current opinion favors an autoimmune basis for psoriasis although the precipitating activating signal(s) within psoriatic plaques remains unknown. Candidate skin autoantigens that have cross-reactive determinants with bacterial antigens include keratins. The immunodysregulation resulting from HIV infection may trigger psoriasis in those genetically predisposed by the Cw*0602 allele. Because CD8 T cells recognize antigen in the context of class I molecules, the identification of a human leucocyte antigen (HLA) class I association in HIV associated psoriasis strengthens the argument for an important role for CD8+ T lymphocytes in the immunopathogenesis of psoriasis. HLA-Cw*0602 could act as a cross-reactive target for cytotoxic T lymphocytes (CTLs) responding to processed peptides from microorganisms. PMID- 10833811 TI - Pneumocystis carinii pneumonia as the first presentation of HIV infection in patients older than fifty. AB - A significant increase in the number of elderly patients first diagnosed with HIV infection at the time of presentation with an AIDS-related opportunistic infection has recently been reported. This suggests a significant delay in the diagnosis of HIV infection. Few data are available describing such cases and their outcome. We restrospectively reviewed records of all elderly patients (> 50 years of age) admitted to a New York City hospital over a 3-year period with confirmed Pneumocystis carinii pneumonia (PCP). The mean age was 57.9 +/- 6.6 years. In 80% (8 of 10 cases), the diagnosis of HIV infection was made at presentation with PCP. The mean CD4 count was 34.2 +/- 39.2/mm3 (1-117/mm3), indicating advanced AIDS. The clinical presentation of PCP was similar to that in younger patients. With prompt and appropriate therapy, a 70% survival rate for this hospitalization was achieved, similar to that reported in younger age groups. The diagnosis of HIV infection was not considered until presentation with PCP at an advanced stage of AIDS in 80% of these elderly patients, thus delaying institution of HIV treatment and counseling. Early consideration of HIV infection in the elderly is of importance because of the rising number of AIDS cases in this age group. PMID- 10833812 TI - Impact of new developments in antiretroviral treatment on AIDS prevention and care in resource-poor countries. AB - Combination antiretroviral treatment (ARV) including protease inhibitors, decreased the morbidity and mortality due to AIDS in the industrialized world. Many obstacles remain before ARVs can be introduced in resource-poor countries: high treatment costs, lack of laboratories to monitor the treatment, weak healthcare systems, and many other competing healthcare needs. The introduction of ARVs in resource-poor countries should be closely monitored. The first priority for the use of ARVs in resource-poor countries is the prevention of mother-to-child transmission of HIV. News about the success of ARV treatment may lead to an increase in unsafe behaviors including a decreased use of condoms. Therefore, prevention efforts should be strengthened; especially the development of an HIV vaccine needs to become a top priority. Funds for ARV treatment cannot come from the already strained healthcare budgets of resource-poor countries. The pressure on politicians and international donor agencies to provide ARVs to resource-poor countries should be used to increase overall healthcare budgets and to improve healthcare services in general. PMID- 10833813 TI - HIV healthcare provider-patient interaction: observations on the process of providing antiretroviral treatment. AB - The meeting during which healthcare providers and newly infected HIV+ patients discuss treatment options is a critical stage in patients' care. We describe 10 interactions between healthcare providers and HIV+ patients at two clinics. Two observers coded each of the 10 interactions for the content of the topics discussed and for communication-relevant behaviors. The interactions followed one of two general patterns, according to whether combination antiretroviral treatment was recommended or deferred. Features of the interactions are described. These features include: (1) the presentation of a single-treatment option, (2) a discussion of whether patients have the will to begin therapy and the skills to adhere to treatment regimens, and (3) organizational pressures to include patients in clinical trials. PMID- 10833814 TI - Hematological complications and quality of life in hospitalized AIDS patients. AB - The relationships among the biological and physiological indicators of cytopenias in AIDS and measures of quality of life are not well characterized. The purpose of this secondary analysis was to determine the relationships among anemia, neutropenia and thrombocytopenia and characteristics of the individual, physiological markers, symptoms, functional status, general health perceptions, and well-being in people with AIDS. The five dimensions of the Wilson and Cleary model of health-related quality of life provided the conceptual model for this study. In addition to descriptive statistics, logistic regression was used to analyze clusters of variables. The sample of 146 hospitalized patients with AIDS had an 85% prevalence of anemia, a 53% prevalence neutropenia and a 33% prevalence of thrombocytopenia. The mean age was 38 years old, 19% were female, 35% were white, 27% had a history of injection drug use and the mean T-helper cell count was 74 mm3. The five dimensions of the Wilson and Cleary model offered significant predictability for anemia only. Patients with higher symptom scores were more likely to have treatable anemia, defined as a hematocrit of < 30%. Treatable anemia was also associated with lower self-care scores and lower T helper cells. Fifty-four percent of the cohort were candidates for colony stimulating factors, while only 17% of those eligible received them. These data suggest suboptimal treatment of anemia and neutropenia in this cohort of AIDS patients. PMID- 10833815 TI - St. John's Wort and HAART. PMID- 10833816 TI - Pill burden key to doing well. PMID- 10833817 TI - ABT-378/r helpful in failing patients. PMID- 10833818 TI - Naltrexone for lipodystrophy. PMID- 10833819 TI - New web site for clinical trials. PMID- 10833820 TI - HCV compound in trials. PMID- 10833821 TI - Survey of teen sex attitudes. PMID- 10833822 TI - Tests may replace Pap smears. PMID- 10833823 TI - Tree bark yields antifungal. PMID- 10833824 TI - Genetics in the John A. Burns School of Medicine Curriculum: a nationally recognized need and an opportunity for curricular integration. PMID- 10833825 TI - [Risk factors in the surgical treatment of patients with lung cancer]. PMID- 10833826 TI - [The optimization of preoperative preparation and the prevention of hemodynamic disorders during anesthesia in middle-aged and elderly patients with concomitant hypertension]. PMID- 10833827 TI - [A trial of using preparations for epidural anesthesia]. PMID- 10833828 TI - [beta-Endorphin in the blood plasma and cerebrospinal fluid--a marker of analgesic efficacy in acute postoperative and chronic pain in cancer patients]. PMID- 10833829 TI - [The effect of anesthesia on lipid peroxidation, the antioxidant system and lipid metabolism during cesarean section in parturients with severe forms of late gestosis]. AB - Forty-three parturients with severe late gestosis were subjected to cesarean section under endotracheal general anesthesia (EGA) (23 pts) and combined epidural anesthesia (CEA) (20 pts). Lipid peroxidation (LPO), antioxidant system (AOS), and lipid metabolism parameters were evaluated at various stages of anesthesia and operation. EN involved a marked increase in LPO, suppressed the activities of vitamin and enzymatic components of AOS, and disturbed lipid metabolism, while CEA was associated with a less pronounced intensification of LPO, slight suppression of AOS, and slight changes in lipid metabolism, which makes this types of analgesia preferable. PMID- 10833830 TI - [Total intravenous anesthesia based on propofol infusion with target concentration: a new procedural solution]. AB - Total intravenous propofol anesthesia with target control (TCI) infusion was used in 61 surgical patients (43 women, 18 men, ASAI-II). Propofol concentration in the central compartment at awakening was in high correlation with the effective concentration (Ce) at the moment of falling asleep (r = 0.64). A model investigation demonstrated the advantages of longer induction for minimization of the hemodynamic disorders. Propofol TCI in a dose of 2 micrograms/ml led to an increase in the EEG spectral potency in the 1-12 Hz range, appearance of biomodality, and a decrease in the right-side frequency. The studies helped determine the policy of intravenous propofol TCI anesthesia using the Ce values at the moment when the patient lost consciousness and the right-side frequency of EEG spectrum. PMID- 10833831 TI - [Structural changes in metabolism in the early postoperative period and the methods for their correction]. AB - Surgery and general anesthesia involve changes in adaptive compensatory mechanisms manifesting by homeostasis disturbances, specifically metabolic disorders. This report describes the dynamic characteristics of structural metabolic disorders and the external respiratory function during the immediate and early postoperative period after surgery of different duration under different anesthesias and metabolic infusions. The results indicate that general anesthesia and any surgery involve structural disorders in energy expenditures during the early postoperative period. The duration of anesthesia and surgery and methods of anesthesia determine the metabolic disturbances and external respiratory changes during the postoperative period. Addition of opioids and hypertensive glucose infusions to general anesthesia and postoperative period decreases the severity of structural metabolic disorders and external respiratory function during the early postoperative period. PMID- 10833832 TI - [Cytokines in the treatment of a generalized surgical infection]. AB - A novel approach to the treatment of surgical sepsis has been tried: cytokine based immunotherapy. Human recombinant interleukin-2 (IL-2) and a mixture of native cytokines obtained by arteriovenous perfusion of xenospleen were used as a source of proinflammatory cytokines. Extracorporeal immunotherapy of 62 patients with surgical sepsis with mononuclear cells treated by autologous IL-2 resulted in a significant decrease of endotoxicosis and effective immunocorrection. Cytokine-based immunotherapy notably decreased the mortality of patients with generalized surgical infection--to 14.6%, which was lower than the expected mortality (35%) and the mortality in the control group (34.5%). PMID- 10833833 TI - [Systemic hemodynamics, oxygenating lung function and oxygen transport in patients with acute intestinal obstruction and an endotoxicosis syndrome]. AB - Systemic hemodynamics, oxygenating function of the lungs, and oxygen budgeting were studied in 32 patients with acute bowel obstruction (ABO) with severe endotoxicosis. Profound disturbances of systemic hemodynamics and blood oxygenation in the lungs underlie the hemostasis changes in this category of patients before and early after operation. These hemostasis changes lead to inadequate oxygen delivery and metabolic requirement of oxygen and are associated with development of oxygen deficiency in tissues. As a rule, patients with ABO with severe endotoxicosis develop acute respiratory failure presenting as adult respiratory distress syndrome on days 2-3 postoperation. Significant differences in the pattern of hemostasis changes and compensatory shifts in survivors and patients dying after ABO are detected at this period, which determine the prognosis and outcome of disease. PMID- 10833834 TI - [An evaluation of the circulatory indices and of the blood oxygen-transport function in relation to the level of endogenous intoxication in patients with abdominal sepsis]. AB - Septic shock in the presence of abdominal sepsis is one of the main cases of death of surgical patients. Hemodynamics was studied by impedancometry in 46 patients with abdominal sepsis. Hemodynamic changes and oxygen transporting function of the blood correlated with the severity of endogenous intoxication. Erythrocyte adsorption capacity was regarded as endogenous intoxication marker. PMID- 10833835 TI - [Multiple-organ pathology in septic shock in patients with hemoblastoses]. AB - A prospective study of 20 patients with hemoblastosis and septic shock (SS) was carried out by invasive monitoring of the central hemodynamics and oxygen transport, evaluation of biochemical and coagulological parameters, and assessment of the severity of clinical condition by the APACHE II and SOFA scores. Septic shock was effectively treated in 12 patients, 5 of them were discharged from the department (group 1) and 7 died in intensive care wards from various complications (group 2). Eight patients died during the first 2 days from SS resistant to therapy (group 3). Group 2 patients were in need of a longer inotropic support than group 1 patients (5.8 +/- 1.6 vs. 2.7 +/- 0.8 days, p < 0.01). The deficit of bases was more expressed in groups 2 and 3 in comparison with group 1 (-11.3 +/- 3 and -2.7 +/- 9.1 mmol/liter vs. 1.4 +/- 4.4 mmol/liter) and left ventricular stroke index (LVSI) and oxygen delivery were lower. LVSI and base deficit were in linear correlation (rho = 0.4, p < 0.05). XIIa-dependent fibrinolysis was suppressed in all patients, which was more pronounced in group 3 in comparison with groups 1 and 2 (135 +/- 47.4 vs. 103 +/- 27 and 88.3 +/- 42.3). According to SOFA score, the severity of cardiovascular disorders during day 1 of SS was the same in all groups, while starting from day 2 it decreased in patients who survived. Acute respiratory failure was lower in group 1 only on day 1 according to SOFA. More pronounced (according to SOFA) hepatorenal failure was observed in group 2 in comparison with other patients. Organ involvement in hemoblastosis was detected at autopsy in 8 out of 13 cases. Hence, the need in prolonged cardiovascular support of SS patients is associated with development of polyorgan involvement. Fibrinolysis suppression is a frequent early manifestation of hemostasis disorders. Specific neoplastic organ involvement was observed in 61.5% patients with hemoblastosis who died from SS and its complications. PMID- 10833836 TI - [Disorders of cardiac contractility caused by hemorrhagic hypotension]. AB - Two-hour hemorrhagic hypotension was induced in outbred male rats. Left ventricular myocardial contractility was studied at all stages of hypotension on a model of isolated isovolumetrically contracting heart. The contractile function decreased after 60 min of hemorrhagic hypotension, this decrease progressing with hypotension prolongation. The myocardial relaxation mechanisms were more involved, and the pathogenetic factors of cardiac depression were hypoergosis, cardiomyocyte membrane injury, and inhibition of Ca(2+)-Mg(2+)-dependent ATPase of sarcoplasmatic reticulum and sarcolemma. PMID- 10833837 TI - [The pathophysiological bases of hyperventilation and its assessment during patient transition from prolonged AV to spontaneous breathing]. PMID- 10833838 TI - [Postural therapy during mechanical pulmonary ventilation with PEEP in patients with unilateral lung damage]. AB - Mechanical ventilation of the lungs (MVL) with positive end expiratory pressure (PEEP) is difficult in patients with unilateral lung damage because of uneven distribution of volumes and pressures in the involved and intact lungs. Harmful effects are easier manifested under such conditions. Selective MVL with selective PEEP is widely used abroad for optimizing MVL, but this method is rather expensive and is not devoid of shortcomings. Our study carried out in 32 patients with unilateral lung involvement showed that traditional MVL with general PEEP can effectively (in 75% cases) regulate gaseous exchange and decrease its untoward effects if MVL is performed with the patient lying on the healthy side and not supine. MVL in patients with unilateral lung injury lying on the healthy side can be a simpler and cheaper alternative to selective MVL with selective PEEP. PMID- 10833839 TI - [The effect of Clexane (low-molecular-weight heparin) on the hemostatic system and on the incidence of postoperative thrombotic complications in gynecological cancer patients]. AB - Clexane in a dose of 40 mg before and after surgery notably decreased the intensity of chronic intravascular blood coagulation during surgical treatment of oncogynecological patients. Only 3% patients developed thrombosis of deep veins of the lower limbs after Clexane, while in the group administered no Clexane, 20% developed this complication. Preventive administration of Clexane did not cause an increase of the operative blood loss. The incidence of wound hematomas in patients administered Clexane and nonfractionated heparin was negligible and virtually the same. PMID- 10833840 TI - [The computer analysis of the variability of the cardiac rhythm: methods, interpretation and clinical use]. PMID- 10833841 TI - [The need and outlook for alkalizing therapy in cardiopulmonary resuscitation]. PMID- 10833842 TI - [Oxygen transport and consumption in patients in critical states]. PMID- 10833844 TI - Analysis of synaptic quantal depolarizations in smooth muscle using the wavelet transform. AB - The time-frequency characteristics of synaptic potentials contain valuable information about the process of neurotransmission between nerves and their target organs. For example, at the synapse between autonomic nerves and smooth muscle, two central issues of neurophysiology, i.e., 1) the probability of neurotransmitter release and 2) the quantal behavior of transmission can be deduced from analysis of the rising phases of evoked excitatory junction potentials (eEJP's) recorded from smooth muscle. eEJP rising phases are marked by prominent inflexions, which reflect these features of neuronal activity. Since these inflexions contain time-varying frequency information, we have applied recent techniques of time-frequency analysis based upon wavelet transforms to eEJP's recorded from the guinea-pig vas deferens in vitro. We find that these techniques allow accurate and convenient characterization of neuronal release sites, and that their probability of release falls between 0.001-0.004. We have also analyzed eEJP's recorded in the presence of the chemical 1-heptanol, which reveals quantal depolarizations. These results have helped clarify the nature of the quantal depolarizations that underly eEJP's. The present method offers significant advantages over those previously employed for these tasks, and holds promise as a novel approach to the analysis of synaptic potentials. PMID- 10833843 TI - [The oxidative modification of blood plasma proteins in patients in critical states]. AB - Current concepts on the formation and biological activity of activated oxygen forms (AOF) in humans are discussed. The main AOF types are produced as a result of consecutive single-electron recovery of molecular oxygen (O2) and are more reactive than O2. AOF are initially normal components of cellular metabolism with certain biological functions. Their reactive aggressiveness is regulated by a potent antioxidant system which is present in any live organism. In disease this balance is distorted towards uncontrolled AOF generation and formation of oxidative stress, when AOF impair all biological structures, including proteins. Unregulated modification of proteins by AOF results in loss of protein biological activities (enzymatic, receptor, transporting function, etc.). Moreover, oxidative modification of proteins generates new antigens and provokes immune response. The authors present experimental data which confirm significant modification of plasma proteins in critical patients. The role of the detrimental effect of AOF on proteins in the formation of critical states deserves special studies. PMID- 10833845 TI - Adaptive whitening of the electromyogram to improve amplitude estimation. AB - Previous research showed that whitening the surface electromyogram (EMG) can improve EMG amplitude estimation (where EMG amplitude is defined as the time varying standard deviation of the EMG). However, conventional whitening via a linear filter seems to fail at low EMG amplitude levels, perhaps due to additive background noise in the measured EMG. This paper describes an adaptive whitening technique that overcomes this problem by cascading a nonadaptive whitening filter, an adaptive Wiener filter, and an adaptive gain correction. These stages can be calibrated from two, five second duration, constant-angle, constant-force contractions, one at a reference level [e.g., 50% maximum voluntary contraction (MVC)] and one at 0% MVC. In experimental studies, subjects used real-time EMG amplitude estimates to track a uniform-density, band-limited random target. With a 0.25-Hz bandwidth target, either adaptive whitening or multiple-channel processing reduced the tracking error roughly half-way to the error achieved using the dynamometer signal as the feedback. At the 1.00-Hz bandwidth, all of the EMG processors had errors equivalent to that of the dynamometer signal, reflecting that errors in this task were dominated by subjects' inability to track targets at this bandwidth. Increases in the additive noise level, smoothing window length, and tracking bandwidth diminish the advantages of whitening. PMID- 10833846 TI - Subspace averaging of steady-state visual evoked potentials. AB - A new algorithm for doing signal averaging of steady-state visual evoked potentials (VEP's) is described. The subspace average is obtained by finding the orthogonal projection of the VEP measurement vector onto the signal subspace, which is based on a sinusoidal VEP signal model. The subspace average is seen to out-perform the conventional average using a new signal-to-noise-ratio-based performance measure on simulated and actual VEP data. PMID- 10833847 TI - Nonstationary time-series analysis applied to investigation of brainstem system dynamics. AB - Previous investigations of the dynamic organization of the lower brainstem and its relation to peripheral and other central nervous systems were predominantly performed by linear methods. These are based on time-averaging algorithms, which merely can be applied to stationary signal intervals. Thus, the current concept of the common brainstem system (CBS) in the reticular formation (RF) of the lower brainstem and basic types of its functional organization have been developed. Here, we present experiments where neuronal activities of the RF and the nucleus tractus solitarii (NTS, first relay station of baroreceptor afferents) were recorded together with related parameters of electroencephalogram (EEG), respiration, and cardiovascular system. The RF neurons are part of the CBS, which participates in regulation and coordination of cardiovascular, respiratory, and motor systems, and vigilance. The physiological time series, thus acquired, yield information about the internal dynamic coordination of the participating regulation processes. The major problem in evaluating these data is the nonlinearity and nonstationarity of the signals. We used a set of especially designed time resolving methods to evaluate nonlinear dynamic couplings in the interaction between CBS neurons and cardiovascular signals, respiration and the EEG, and between NTS neurons (influenced by baroreceptor afferents) and CBS neurons. PMID- 10833848 TI - Decomposition of posterior alpha rhythm. AB - The classical posterior alpha rhythm has been decomposed into regular and irregular components using orthogonal transformation. A periodicity generator is considered, which has three characteristic control parameters: the periodicity, the amplitudes or scaling factors and the pattern associated with successive periodic segments. The regular component is shown to be equivalent to an oscillator or periodicity generator whose parameters are dynamically varying and, thus, producing both amplitude- and frequency-modulation. The irregular component is devoid of such modulating behavior. Electroencephalogram signals from normal, maniac and epileptic subjects are studied. Through analytic signal-based analysis, it is shown that for the regular component, healthy brain possesses universal scaling behavior, whereas heterogeneous scaling or absence of universality is observed for the diseased brain. PMID- 10833849 TI - Uterine EMG analysis: a dynamic approach for change detection and classification. AB - Toward the goal of detecting preterm birth by characterizing events in the uterine electromyogram (EMG), we propose a method of detection and classification of events in this signal. Uterine EMG is considered as a nonstationary signal and our approach consists of assuming piecewise stationarity and using a dynamic change detector with no a priori knowledge of the parameters of the hypotheses on the process state to be detected. The detection approach is based on the dynamic cumulative sum (DCS) of the local generalized likelihood ratios associated with a multiscale decomposition using wavelet transform. This combination of DCS and multiscale decomposition was shown to be very efficient for detection of both frequency and energy changes. An unsupervised classification based on the comparison between variance-covariance matrices computed from selected scales of the decomposition was implemented after detection. Finally a class labeling based on neural networks was developed. This algorithm of detection-classification labeling gives satisfactory results on uterine EMG: in most cases more than 80% of the events are correctly detected and classified whatever the term of gestation. PMID- 10833850 TI - A new and fast nonlinear method for association analysis of biosignals. AB - In this paper, we present some original theoretical aspects of a fast nonlinear association measure based on the work of Cramer. The features of this new measure -the V measure--when applied to biosignals are also shown using simulated time series. A comparative study with other well-known association measures available in the literature of biosignals is presented. V was found to be twice as fast and more robust to nonlinearities than the classical cross-correlation ratio (r2) and more than 100 times faster than the nonlinear regression coefficient (h2), presenting similar behavior in the presence of nonlinear simulated situations. This new measure is very fast and versatile. It is appropriate to deal with nonlinear relations presenting usually a sharp peak in the association function enabling a high degree of selectivity for maxima detection. It seems to constitute an improvement over linear methods of association which is faster and more robust to the existing nonlinearities. It can be used as an alternative to more complex nonlinear association measures when computational speed is an important feature. PMID- 10833851 TI - A new approach for TU complex characterization. AB - In this paper, we present a new TU complex detection and characterization algorithm that consists of two stages; the first is a mathematical modeling of the electrocardiographic segment after QRS complex; the second uses classic threshold comparison techniques, over the signal and its first and second derivatives, to determine the significant points of each wave. Later, both T and U waves are morphologically classified. Amongst the principal innovations of this algorithm is the inclusion of U-wave characterization and a mathematical modeling stage, that avoids many of the problems of classic techniques when there is a low signal-to-noise ratio or when wave morphology is atypical. The results of the algorithm validation with the recently appeared QT database are also shown. For T waves these results are better when compared to other existing algorithms. U-wave results cannot be contrasted with other algorithms as, to our knowledge, none are available. Examples showing the causes of principal discrepancies between our algorithm and the QT database annotations are also given, and some ways of attempting to improve and benefit from the proposed algorithm are suggested. PMID- 10833852 TI - Adaptive time-frequency analysis of knee joint vibroarthrographic signals for noninvasive screening of articular cartilage pathology. AB - Vibroarthrographic (VAG) signals emitted by human knee joints are nonstationary and multicomponent in nature; time-frequency distributions (TFD's) provide powerful means to analyze such signals. The objective of this paper is to construct adaptive TFD's of VAG signals suitable for feature extraction. An adaptive TFD was constructed by minimum cross-entropy optimization of the TFD obtained by the matching pursuit decomposition algorithm. Parameters of VAG signals such as energy, energy spread, frequency, and frequency spread were extracted from their adaptive TFD's. The parameters carry information about the combined TF dynamics of the signals. The mean and standard deviation of the parameters were computed, and each VAG signal was represented by a set of just six features. Statistical pattern classification experiments based on logistic regression analysis of the parameters showed an overall normal/abnormal screening accuracy of 68.9% with 90 VAG signals (51 normals and 39 abnormals), and a higher accuracy of 77.5% with a database of 71 signals with 51 normals and 20 abnormals of a specific type of patellofemoral disorder. The proposed method of VAG signal analysis is independent of joint angle and clinical information, and shows good potential for noninvasive diagnosis and monitoring of patellofemoral disorders such as chondromalacia patella. PMID- 10833853 TI - Modeling the cardiac action potential using B-spline surfaces. AB - This paper presents a new method for constructing empirical, two-state-variable models of cardiac cell membrane kinetics. The formulation is based on nonuniform rational B-spline surfaces that can be manipulated interactively to produce desired action potential (AP) properties. Using this new methodology, a model of the guinea pig ventricular action potential was constructed that reproduces experimentally measured relationships between pacing cycle length and action potential duration and conduction velocity. The model is computationally efficient, requiring about sixfold less CPU time than the Beeler-Reuter ionic model and only about twice as much time as a FitzHugh-Nagumo type empirical model. Thus, for modeling propagation phenomena, this method can produce models that improve on the quantitative accuracy of both simple empirical models and elaborate ionic models, with computational cost comparable to the simplest of empirical models. PMID- 10833854 TI - The boundary element method in the forward and inverse problem of electrical impedance tomography. AB - In this paper, a new formulation of the reconstruction problem of electrical impedance tomography (EIT) is proposed. Instead of reconstructing a complete two dimensional picture, a parameter representation of the gross anatomy is formulated, of which the optimal parameters are determined by minimizing a cost function. The two great advantages of this method are that the number of unknown parameters of the inverse problem is drastically reduced and that quantitative information of interest (e.g., lung volume) is estimated directly from the data, without image segmentation steps. The forward problem of EIT is to compute the potentials at the voltage measuring electrodes, for a given set of current injection electrodes and a given conductivity geometry. In this paper, it is proposed to use an improved boundary element method (BEM) technique to solve the forward problem, in which flat boundary elements are replaced by polygonal ones. From a comparison with the analytical solution of the concentric circle model, it appears that the use of polygonal elements greatly improves the accuracy of the BEM, without increasing the computation time. In this formulation, the inverse problem is a nonlinear parameter estimation problem with a limited number of parameters. Variants of Powell's and the simplex method are used to minimize the cost function. The applicability of this solution of the EIT problem was tested in a series of simulation studies. In these studies, EIT data were simulated using a standard conductor geometry and it was attempted to find back this geometry from random starting values. In the inverse algorithm, different current injection and voltage measurement schemes and different cost functions were compared. In a simulation study, it was demonstrated that a systematic error in the assumed lung conductivity results in a proportional error in the lung cross sectional area. It appears that our parametric formulation of the inverse problem leads to a stable minimization problem, with a high reliability, provided that the signal-to-noise ratio is about ten or higher. PMID- 10833855 TI - Maximum-likelihood versus maximum a posteriori parameter estimation of physiological system models: the C-peptide impulse response case study. AB - Maximum-likelihood (ML), also given its connection to least-squares (LS), is widely adopted in parameter estimation of physiological system models, i.e., assigning numerical values to the unknown model parameters from the experimental data. A more sophisticated but less used approach is maximum a posteriori (MAP) estimation. Conceptually, while ML adopts a Fisherian approach, i.e., only experimental measurements are supplied to the estimator, MAP estimation is a Bayesian approach, i.e., a priori available statistical information on the unknown parameters is also exploited for their estimation. In this paper, after a brief review of the theory behind ML and MAP estimators, we compare their performance in the solution of a case study concerning the determination of the parameters of a sum of exponential model which describes the impulse response of C-peptide (CP), a key substance for reconstructing insulin secretion. The results show that MAP estimation always leads to parameter estimates with a precision (sometimes significantly) higher than that obtained through ML, at the cost of only a slightly worse fit. Thus, a three exponential model can be adopted to describe the CP impulse response model in place of the two exponential model usually identified in the literature by the ML/LS approach. Simulated case studies are also reported to evidence the importance of taking into account a priori information in a data poor situation, e.g., when a few or too noisy measurements are available. In conclusion, our results show that, when a priori information on the unknown model parameters is available, Bayes estimation can be of relevant interest, since it can significantly improve the precision of parameter estimates with respect to Fisher estimation. This may also allow the adoption of more complex models than those determinable by a Fisherian approach. PMID- 10833856 TI - Micromachined pipette arrays. AB - In this paper, the design and characterization of batch fabricated metallic micromachined pipette arrays is described. The process used to fabricate the micromachined pipette arrays (MPA) includes p+ etch-stop membrane technology, anisotropic etching of silicon in potassium hydroxide, sacrificial thick photoresist micromolding technology, and electrodeposition. Arrays of one to ten pipettes have been fabricated using nickel as the structural material and palladium as the biocompatible coating of inside walls. The inner dimensions of the individual pipettes fabricated to date range from 30 microns to 1.5 mm in width, 0.5 mm to several cm in length, and 5-50 microns in thickness. The center to-center spacing of these pipettes varies from 100 microns to several centimeters. The MPA have a number of advantages when compared to the current micropipette technology, including the ability to transfer precise volumes of samples in the submicroliter range; the ability to manipulate samples, reagents, or buffers in a highly-parallel fashion by operating hundreds of individual pipettes simultaneously; and the compatibility with the submilimeter center-to center dimensions of the microscale biochemical analysis systems. The application of the MPA to high lane density slab gel electrophoresis is explored. Sample wells are formed in agarose gels by using micromachined combs (solid MPA) at center-to-center spacing ranging from 250 microns to 1.9 mm. The samples are loaded using the MPA. The results of the micro-gel separations compare favorably with the standard mini-gel separations and show a twofold increase in the number of theoretical plates as well as a sixfold increase in lane density. PMID- 10833857 TI - An S1 gradient of refractoriness is not essential for reentry induction by an S2 stimulus. AB - This communication reports a numerical simulation of reentry induction by successive stimulation (S1-S2) that does not require an S1 gradient of refractoriness. The S1 action potential is uniform in space, so before the S2 stimulus there is no refractory gradient. Nevertheless, a unipolar S2 stimulus initiates quatrefoil reentry. The result supports the growing realization that virtual electrodes, hyperpolarization, de-excitation, and break excitation may be important during reentry induction. PMID- 10833858 TI - Overcoming selective ensemble averaging: unsupervised identification of event related brain potentials. AB - We present a novel approach to the problem of event-related potential (ERP) identification, based on a competitive artificial neural network (ANN) structure. Our method uses ensembled electroencephalogram (EEG) data just as used in conventional averaging, however without the need for a priori data subgrouping into distinct categories (e.g., stimulus- or event-related), and thus avoids conventional assumptions on response invariability. The competitive ANN, often described as a winner takes all neural structure, is based on dynamic competition among the net neurons where learning takes place only with the winning neuron. Using a simple single-layered structure, the proposed scheme results in convergence of the actual neural weights to the embedded ERP patterns. The method is applied to real event-related potential data recorded during a common odd-ball type paradigm. For the first time, within-session variable signal patterns are automatically identified, dismissing the strong and limiting requirement of a priori stimulus-related selective grouping of the recorded data. The results present new possibilities in ERP research. PMID- 10833859 TI - Cosmetic dentistry: is it really health care? PMID- 10833860 TI - Accepting the need for "overkill" in infection control. PMID- 10833862 TI - Life insurance that's a custom fit. PMID- 10833861 TI - Dentistry isn't about selling! PMID- 10833863 TI - Internet security. PMID- 10833864 TI - Managed care plans on campuses. PMID- 10833865 TI - The building blocks of evidence-based dentistry. AB - The practice of dentistry is becoming more complex and challenging. Changing socio-demographic patterns, patients as knowledgeable health care consumers, rapid technical advances and the information "explosion" all place greater demands on clinical decision making. The need for reliable information and the electronic revolution have come together to allow the "paradigm shift" towards evidence-based health care to progress swiftly. Evidence-based dentistry closes the gap between clinical research and real world dental practice and provides dentists with powerful tools to interpret and apply research findings. Central to evidence-based practice is the systematic literature review, which synthesizes the best evidence and provides the basis for clinical practice guidelines. PMID- 10833866 TI - "Free" dental care for children. PMID- 10833867 TI - The new business ethics. PMID- 10833868 TI - Tobacco-associated lesions of the oral cavity: Part I. Nonmalignant lesions. AB - The excessive use of tobacco products has been associated with various lesions in the oral cavity. Tobacco-associated lesions include tooth stains, abrasions, smoker's melanosis, acute necrotizing ulcerative gingivitis and other periodontal conditions, burns and keratotic patches, black hairy tongue, nicotinic stomatitis, palatal erosions, leukoplakia, epithelial dysplasia and squamous-cell carcinoma. A routine intraoral examination by a dental health professional can reveal most of these lesions at an early stage, and early intervention may prevent serious sequelae. Thus, detection of tobacco-associated lesions in conjunction with tobacco-use counselling by dental professionals has become the standard of care. The significance and treatment of nonmalignant tobacco associated lesions are addressed in the first part of this two-part series. Malignant lesions, basic biopsy principles and prevention and intervention strategies are discussed in the subsequent part. PMID- 10833869 TI - Examination, classification, and treatment of halitosis; clinical perspectives. AB - Patients with halitosis may seek treatment from dental clinicians for their perceived oral malodour. In this article, an examination protocol, classification system and treatment needs for such patients are outlined. Physiologic halitosis, oral pathologic halitosis and pseudo-halitosis would be in the treatment realm of dental practitioners. Management may include periodontal or restorative treatment or both, as well as simple treatment measures such as instruction in oral hygiene, tongue cleaning and mouth rinsing. Psychosomatic halitosis is more difficult to diagnose and manage, and patients with this condition are often mismanaged in that they receive only treatments for genuine halitosis, even though they do not have oral malodour. A classification system can be used to identify patients with halitophobia. Additionally, a questionnaire can be used to assess the psychological condition of patients claiming to have halitosis, which enables the clinician to identify patients with psychosomatic halitosis. In understanding the different types of halitosis and the corresponding treatment needs, the dental clinician can better manage patients with this condition. PMID- 10833870 TI - Computed tomographic imaging of the foot and ankle. Developmental and congenital anomalies. AB - Computed tomography is a primary imaging technique for evaluating congenital and developmental anomalies of the foot and ankle. Other imaging modalities have special capabilities, but computed tomography is a fast, safe, and effective method of assessing the anatomy. To demonstrate this point, the authors present and discuss imaging findings of common anomalies of the foot and ankle. PMID- 10833871 TI - The Boyd amputation as a treatment for osteomyelitis of the foot. AB - The Boyd amputation is a surgical technique used to treat osteomyelitis of the foot. This amputation is a technically more difficult procedure to perform than the Syme amputation, but it offers certain advantages. The Boyd amputation provides a more solid stump because it preserves the function of the plantar heel pad. Also, because a portion of the calcaneus is left and fused to the tibia, the weightbearing surface is more solid than in the case of a Syme amputation. The authors recommend a Boyd amputation as an alternative to a Syme or a below-the knee amputation to treat patients with osteomyelitis of the forefoot and midfoot. PMID- 10833872 TI - Use of small cannulated screws for fixation in foot surgery. AB - Use of cannulated bone screws, as compared with use of traditional bone screws, has been reported to decrease surgical time, allow for more precise screw placement, and reduce sources of error. Cannulation of the smaller-size screws that are routinely used in foot surgery has not been available until the last few years. This article reports on the use of the small cannulated screws manufactured by Alphatec Manufacturing, Inc (Palm Desert, California). The screw sizes available in the Mini Lag Screw System are 2.7, 3.5, and 4.0 mm. A long term clinical and radiographic prospective evaluation of 70 procedures performed on 49 patients was conducted. The follow-up time for all patients was 2 years. None of the 70 implants fractured, and seven procedures (in seven patients) resulted in some type of implant-fixation failure. All of the fixation failures, however, appeared to be related to an untoward event or patient noncompliance. These smaller cannulated screws proved to be a reliable and effective means of fixation in foot surgery. PMID- 10833873 TI - Comparison of three types of postoperative management for endoscopic plantar fasciotomy. A retrospective study. AB - This study compared three types of postoperative management of patients undergoing endoscopic plantar fasciotomy. Participating in the study were 38 patients (45 feet) who had undergone endoscopic plantar fasciotomy from 1995 to 1998. One group of patients wore a below-the-knee walking cast with a molded medial longitudinal arch for 2 weeks; another group was allowed immediate plantigrade weightbearing; the third group was nonweightbearing with crutch assisted ambulation for 2 weeks. The results showed that the patients who wore the below-the-knee walking cast for 2 weeks required less time to obtain 80% pain relief, needed less time to return to full activities, and had fewer complications than those patients who were allowed immediate plantigrade weightbearing. Moreover, patients who wore the below-the-knee walking cast were more satisfied with their postoperative results than patients who were nonweightbearing for 2 weeks. PMID- 10833874 TI - Multiple neuromas coexisting with rheumatoid synovitis and a rheumatoid nodule. AB - The authors present a rare case of multiple intermetatarsal neuromas coexisting with rheumatoid synovitis and a rheumatoid nodule. A brief review of rheumatoid nodules as a source of forefoot pain and a review of the relevant literature are provided. A rheumatoid nodule is just one of the many diagnoses that must be considered when one encounters pedal symptoms similar to those associated with Morton's neuroma. PMID- 10833875 TI - Origin of foot ganglion. PMID- 10833876 TI - A conceptual model for a geriatric syllabus for podiatric medicine. AB - Foot disorders and the complications of chronic disease in the older population have a significant effect on society, the cost of health care, and individuals' quality of life. Given podiatric medicine's role in the management of problems of the aging, it is critical that the profession's educational system produce practitioners who understand the process of aging and the needs of the older patient and who are prepared to serve on the health-care-delivery team as the primary providers of foot-care services. The geriatric syllabus presented here is one approach to attaining that goal. PMID- 10833877 TI - Minocycline-induced cutaneous hyperpigmentation. PMID- 10833878 TI - Giant cell tumor of tendon sheath in the toe. PMID- 10833879 TI - Disseminated gonococcal infection: a case of beta-lactamase-producing Neisseria gonorrhoeae. PMID- 10833880 TI - [Gastropathy induced by nonsteroidal anti-inflammatory drugs (Part II)]. PMID- 10833881 TI - [New approaches to diet therapy for rheumatic diseases]. PMID- 10833882 TI - [Autonomous nervous system in patients with early postmyocardial infarction angina pectoris]. AB - Ninety-three patients with acute myocardial infarction were examined. They were divided into 2 groups: 1) those with early postmyocardial infarction angina pectoris and 2) those without it. The autonomic nervous system was studied by exploring its segmental and suprasegmental regions. The autonomic tone was evaluated by variance intervalometry. The test of human lymphocytic rosette formation and bovine red blood cells sensitized with glutaric aldehyde and loaded epinephrine was applied to estimate the adrenoception. The sympathetic tone and tissue adrenoceptor density were found to be higher in patients with early postinfarction angina than in those without it. The finding may be useful in predicting early postinfarction angina in patients with acute myocardial infarction. PMID- 10833883 TI - [On possible prognostic values of anti-endotoxin immunological parameters in acute myocardial infarction]. AB - Endotoxin is a biologically active substance that has a lipopolysaccharide structure. It is found in the cell walls of microorganisms, principally gram negative bacteria. By contacting with the cell, endotoxin enhances immunity. The findings suggest that acute myocardial infarction (AMI) occurs due to low immunity that remains the same at week 3 of the disease, thus the levels of antiendotoxin agents may be used in predicting AMI. PMID- 10833884 TI - [Antiaggregation activity of the vascular wall and pancreatic ultrasound pattern in the development of chronic alcohol-induced pancreatitis]. AB - There are ultrasonographic signs of pancreatic edematous changes in early chronic alcohol-induced pancreatitis and signs of fibrous changes in the advanced (over 10-year) disease. Progressive chronic alcohol-induced pancreatitis is followed by decreased vascular wall antiaggregatory activity more commonly detected in pancreatic tissue fibrous changes, which should be borne in mind in choosing the optimum treatment policy for the patient. PMID- 10833885 TI - [Diagnosis of the hibernating myocardium in early stages of heart failure in patients with coronary heart disease]. AB - The paper presents current views on ischemia-reperfusion and mechanisms of its development. Their role in the development of the hibernating myocardium is shown. Possible clinical manifestations of these conditions and currently available diagnostic techniques are assessed. The purpose of the study was to detect the hibernating myocardium in patients with coronary heart disease (CHD) and arterial hypertension (AH). The study enrolled 13 patients with CHD and AH. Myocardial biventricular scintigraphy was conducted in patients before treatment and during acute coronarolytic test. Prior to treatment, all the patients were found to have reversible asynergy, left and right ventricular myocardial hypo- and dyskinesia caused by the hibernating myocardium. The drug test revealed better segmental contractility than the baseline one due to the fact that the function of asynergic segments recovered and fraction ejection increased. The findings suggest that the hibernating myocardium is diagnosed in patients with CHD and AH without myocardial infarction, which is of importance in determining treatment policy and prognosis in these patients. PMID- 10833886 TI - [High-density lipoproteins in cholesterosis of the gall bladder]. AB - The composition of serum high-density lipoproteins (HDL) was studied in 64 patients with polypous cholesterosis (PC). The spectrum of serum lipids in patients with PC was characterized by the lower concentrations of HDL cholesterol (42.0 +/- 2.5 mg/dl; p < 0.05) and higher concentrations of low-density lipoproteins (LDL) cholesterol (169.9 +/- 6.9 mg/dl; p < 0.01) than those in the controls. The decreased HDL cholesterol, or hypoalphacholesterolemia was associated with quantitative changes in HDL phospholipids (PL) (66.48 +/- 3.4; p < 0.01) and with changes in the composition of individual PL by lowering the proportion of lecithin (47.13 +/- 2.19 mg/dl; p < 0.01). It may be suggested that the lower amount of HDL cholesterol is caused by the decreased HDL acception of free cholesterol from the peripheral cell membranes due to the impaired complexation of PL with free cholesterol and associated the altered PL composition of the superficial monolayer of a lipoprotein particle. At the same time the physicochemical changes in Hdl superficial layer are a cause of abnormal free cholesterol esterification and the impaired plunge of esterified cholesterol into the nucleus of a HDL particle, which facilitates the conversion of HDL to LDL and may explain elevated LDL levels in cholesterosis. The findings suggest that serum lipids are involved in the development of cholesterosis. PMID- 10833887 TI - [Serum crystalloscopic study in the diagnosis of chronic pancreatitis concurrent with biliary diseases]. AB - The sera from 20 patients with chronic pancreatitis concurrent with diseases of biliary tract and gallbladder were first crystalloscopically studied. Hypertrypsinemia was followed by the appearance of a specific structure appeared as crystals made by a continuum dendritic large-porous lattice. The test proposed is easy-to-use and may be useful in diagnosing chronic pancreatitis. PMID- 10833888 TI - [Possible mechanisms of development of biliary diseases under the conditions of the Transbaikal region]. AB - Eighty one patients with various biliary diseases and 36 apparently healthy individuals were examined. There were relatively elevated lipid peroxidation (LPO) products (E 232/220 and E 278/220) in biliary diskynesia and higher concentrations of LPO intermediate products in cholecystitis. In cholelithiasis, the levels of LPO products were lower than those in cholecystitis. E 232/220 and 278/220 may be used for early diagnosis of biliary dysfunctions. Selenium deficiency observed in the Transbaikal region decreases antiradical protection and increases the incidence of biliary diseases. PMID- 10833889 TI - [Clinical characteristics and risk factors of hepatic damage in lyme borrheliosis]. AB - The study is based on the study of data on 33 patients with Lyme Borrelia infection in the presence of typical erythema migrans in whom elevated levels of serum bilirubin or transaminases were detected simultaneously with erythema or just shortly. The obligatory criterion was no history evidence of hepatitis and abnormal hepatic functional tests. Higher levels of serum aminotransferases were a major manifestation of Lyme hepatitis in the Sverdlovsk region. In 32 patients, ALT was increased, on the average, up to 176 U/l, and AST activity was up to 113 U/l within the first 2 weeks of the disease in the absence of clinical manifestations of hepatic and biliary diseases. There were changes in the levels of serum transaminases and bilirubin following 3- and 8-month antibiotic therapy. The presence of viruses A and C in moderate chronic hepatitis induced long-term increases in the activity of transaminases in 3 cases, as evidenced by histological studies of hepatic biopsy specimens. PMID- 10833890 TI - [Effect of moxonidine monotherapy and in combination with melatonin on hemodynamic parameters in patients with arterial hypertension]. AB - Fifty patients (mean age 53.8 +/- 3.45) with arterial hypertension were examined. They were divided into 2 randomized groups, 25 patients each. One group received moxonidine, 40 mg, alone in the morning, the other had moxonidine in the same dose in combination with melatonin, 3 mg, overnight. The treatment lasted 14 days. Monitoring of blood pressure (BP), heart rate (HR), EchoCG was made before and after treatment. The studies indicated that moxonidine had a pronounced antihypertensive effect by slightly lowering BP overnight and that the chronostructure of BP and HR rhythms was impaired. Addition of melatonin to the moxonidine therapy with greater antihypertensive effects made the chronostructure of circadian BP and HR rhythms normal. PMID- 10833891 TI - [Cardiological use of mononitrates]. AB - The study has provided strong evidence for the marked antihypertensive and antiischemic effects of isosorbide-5-mononitrate (I-5-M)--the drug Monochinque retard, 50 mg (A Monarini Group, Berlin Chemie). It contributed to the reduction of the number of anginal attacks and to the alleviation of myocardial ischemia, as evidenced by Holter monitoring. The findings also indicate that there are positive changes in central hemodynamic parameters, such as left ventricular systolic and diastolic functions when Monochinque was given. The data presented suggest that Monochinque, 50 mg, an I-5-M has a positive effect on the clinical manifestations of pain and painless forms of coronary heart disease and that it alleviated functional classes II-IV heart failure. PMID- 10833892 TI - [Osteoporosis as a problem in internal medicine]. PMID- 10833893 TI - [Bacterial endocarditis in elderly and senile patients]. PMID- 10833894 TI - [Specific features in the course, treatment and prevention of hypertensive crises in the elderly]. PMID- 10833895 TI - [The problems of health and normalcy: a point of view of a physiologist]. PMID- 10833896 TI - [Significant dates and anniversaries in the history of medicine in 2000]. PMID- 10833897 TI - Fundic gland polyps: a still elusive entity on the eve of the year 2000. AB - Fundic gland polyps (FGPs) are tiny multiple sessile polyps of the acid-secreting gastric mucosa. They have been described both in a sporadic form, mainly in middle-aged females, and in a syndromic form, associated with familial adenomatous polyposis (FAP)-Gardner's syndrome and attenuated variants (AFAP). They share the same histology, characterised by superficial and deep cystic dilatations, shortened gastric pits, with an inconspicuous lamina propria. They have been for a long time described as innocuous lesions, but some recent reports have shown that FGPs may harbour dysplastic foci and ultimately (particularly syndromic polyps) gastric cancer. Factors influencing their genesis are unknown. A circulating factor in FAP patients has been postulated and a role of female hormones has been suggested for sporadic FGPs. Whereas patients with sporadic FGPs have normal basal acid output, normal fast serum levels of gastrin and pepsinogen I, the role of gastrin seems crucial for the development of cystic changes in flat body-fundus mucosa, and for the appearance of FGPs in patients with Zollinger-Ellison syndrome. A role of H. pylori induced gastritis has been excluded. Actually, patients with both sporadic and syndromic FGPs appear consistently free from H. pylori colonisation, again for an unknown factor(s). Some recent reports have claimed a role for omeprazole in the genesis of FGPs, a highly controversial issue. Ultimately, the nature of FGPs is still debated: some have interpreted them as hamartomatous lesions, others as a peculiar form of hyperplastic polyp. PMID- 10833898 TI - Biochemical and morphological study on liver Golgi complex in streptozotocin diabetic and control rats treated with bis(kojato)oxovanadium(IV) [VO(ka)2]x2H2O. Part I. One week treatment with vanadium compound. AB - In comparison with untreated control, reduced body and liver weights were found in two groups of rats (such as control and STZ-diabetic) treated orally with bis(kojato)oxovanadium(IV) solution. Free blood sugar in STZ-diabetic rats was lower by about 38%, but did not achieve euglycemic values. Yields of Golgi-rich fraction were lower than those in untreated controls, similar to the activity of galactosyl transferase (GalT) in both vanadium treated groups (control and diabetic). Under electron microscope in the control kojate-treated group, subcellular changes were observed. The morphology of Golgi apparatus was typical, resembled that in untreated animals. In diabetic animals treated with kojate subcellular changes were less severe. Golgi apparatus was usually semicircular or arched in shape similar to that observed previously in diabetic untreated rats. PMID- 10833899 TI - Biochemical and morphological study on liver Golgi complex in streptozotocin diabetic and control rats treated with bis(kojato)oxovanadium(IV) [VO(ka)2]x2H2O. Part II. Prolonged treatmentwith vanadium compound. AB - In this study oral treatment with bis(kojato)oxovanadium(IV) solution was administered twice. The first, short vanadium treatment (so called pretreatment) was used to accustom animals to the flavour of this liquid. After 2-2.5 weeks the second treatment, in high concentration and for a longer time served as a "drug". The physiological parameters such as weights of animals and their livers, reduction of liquid and food intake were lower than in control. Activity of Golgi marker enzyme GalT was significantly lower in both vanadium treated groups (p < 0.01), but in diabetic vanadium treated group it was higher, albeit not significantly, than in control vanadium treated rats. The morphology of Golgi complexes in diabetic rats on prolonged vanadium treatment was similar to that in the one week treated rats (see Part I). Rounded stacks of cisternae characteristic of untreated streptozotocin (STZ)-diabetes were also seen in vanadium treated diabetic rats, but in comparison with untreated diabetic livers, the secretory activities of Golgi complexes were preserved or even stimulated. PMID- 10833900 TI - Endoscopical and histological aspects of inflammatory changes in J-pouch mucosa. AB - The aim of the study was endoscopic and histological evaluation of mucosal inflammatory changes in the ileal reservoir after restorative proctocolectomy with ileoanal anastomosis (IPAA) because of FAP (27 patients) and ulcerative colitis (34 patients). Endoscopic symptoms of pouchitis (edema, granularity, contact bleeding, loss of vascular pattern, mucus, erosions) were found in 13 patients (4 were operated on because of FAP, 9 because of UC). A statistically significant difference (p < 0.0001) was found between endoscopic inflammatory index in UC (4.0 +/- 0.5) and in FAP (1.00 +/- 0.00). The endoscopic changes were correlated with the Moskowitz histological classification, in 12 acute score and in 6 chronic score were above 4 points. In 18 patients histological diagnosis of pouchitis was made (5 in FAP, 13 in UC). Acute pouchitis occurred in 12 patients and chronic in 6. In one patient histological changes resembled Crohn's disease. Regular endoscopic control with histological examination and verification of inflammatory lesions in the ileal reservoir is necessary. PMID- 10833901 TI - Ki-67 antigen and P53 protein expression in benign and malignant prostatic lesions. Immunohistochemical quantitative study. AB - Ki-67 and P53 expression were studied using immunohistochemistry on tissue samples obtained during transurethral electroresection or needle biopsy in 62 patients with prostatic lesions: group 1 (n = 15)--benign prostatic hyperplasia (BPH), group 2 (n = 10)--high-grade prostatic intraepithelial neoplasia (PIN 3), group 3 (n = 10)--low-grade prostatic carcinoma (PC, Gleason score 2-4), group 4 (n = 12) intermediate-grade prostatic carcinoma (PC, Gleason score 5-7) and group 5 (n = 15) high-grade prostatic carcinoma (PC, Gleason score 8-10). Moreover, in the groups examined the associations between expression of Ki-67 and P53 were analysed. Paraffin-embedded tissue samples were immunostained with monoclonal antibody anti-P53 and polyclonal antibody anti-Ki-67 using avidinbiotin peroxidase method. Our study revealed lack of Ki-67 and P53 immunoreactivity in BPH. Only 3 out of 10 high-grade PIN exhibited Ki-67 positivity, but there was no immunopositivity of P53 protein in this group. Although immunopositivity of Ki-67 increased with the histological grade of prostatic cancer, the differences in Ki 67 expression between intermediate and high-grade cancer did not reach statistical significance. A similar level of Ki-67 reactivity in intermediately differentiated and poorly-differentiated prostate cancer suggests a similar biology of these cancers. P53 protein positivity was noted in 62.2% cases of prostate cancer. Moreover, the highest level of P53 accumulation in intermediate grade carcinomas may predict the aggressive progression and risk of metastases in these cases. No significant differences in P53 immunopositivity between low-grade and high-grade PC were noted. Interestingly, only in low-grade PC there was a significant positive correlation between expression of Ki-67 and P53 protein. PMID- 10833902 TI - A quantitative study of the interstitial expression of alpha-smooth muscle actin (alpha-SMA) in idiopathic membranous glomerulonephritis and minimal change disease in adults. AB - Twelve adult patients with early stages of idiopathic membranous glomerulonephritis (MGN) and 10 with minimal change disease (MCD) for whom both light and electron microscopy as well as immunofluorescence microscopy and full clinical data were available were examined quantitatively. Morphometric investigations were performed by means of a computer image analysis system to evaluate whether morphometric analysis could reveal differences between interstitial expression of alpha-smooth muscle actin (alpha-SMA) in early cases of MGN and MCD in adults, and, in particular, to examine the relationship between alpha-SMA expression and interstitial infiltrates in these glomerulopathies. The morphometric study revealed that mean values of the interstitial expression of alpha-SMA, interstitial volume, CD45RB+, CD43+, CD20+ and CD68+ cells were in MGN patients increased in comparison with normal controls, and MCD group, most of them significantly. The correlations between interstitial expression of alpha-SMA and interstitial volume, CD45RB+, CD43+, CD20+ as well as CD68+, cells were in MCD patients weak and not significant. In MGN group significant positive correlations were found between interstitial expression of alpha-SMA and interstitial volume as well as CD68+ cells. In conclusion, our study demonstrated that in adults, interstitial expression of alpha-SMA in early cases of membranous glomerulonephritis, was significantly stronger than that in minimal change disease. It is noteworthy that in MCD interstitial alpha-SMA staining was similar to that in normal kidney. The significant positive correlation between interstitial expression of alpha-SMA and interstitial CD68+ cells needs further investigations to elucidate whether monocytes/macrophages play a role in the process of inducing the myofibroblast phenotype in resting fibroblasts. PMID- 10833903 TI - Significance of DNA ploidy measurements in Spitz nevi. AB - In 28 Spitz nevi DNA content was determined by video-imaging cytometry. The nevi were selected for this study because of difficulties in differentiation from melanoma and heterogeneity of this type of nevus. Morphological features of Spitz nevi and differences helpful for differentiation between Spitz nevi and malignant melanoma were identified. DNA ploidy was measured in paraffin embedded and fresh tissue material from each patient and the results were comparable. The sample preparation process and video-imaging method are presented in this study. Twenty two (78.6%) lesions contained diploid cell populations, 5 (17.9%) aneuploid and 1 (3.6%) tetraploid cell population. A significant correlation was observed between DNA ploidy measured in fresh tissue and retrospective material. The results indicate the presence of abnormal DNA content in some of the lesions. This observation does not indicate that DNA ploidy cytometry is useful for the differentiation of Spitz nevi from malignant melanoma. PMID- 10833904 TI - Hypertrophic gastritis, primary diffuse G-cell hyperplasia and pancreatic metaplasia of the gastric mucosa (pseudo-Zollinger-Ellison syndrome)--case report. AB - We report a case of pseudo-Zollinger-Ellison syndrome in a 17-year-old man presenting with gastrin levels exceeding 2000 pmol/l and BAO 24 mEq/hr. Histologically, apart from hypertrophic gastritis with the thickening of mucosal folds and diffuse G-cell hyperplasia, gastric mucosa was found to contain foci of pancreatic metaplasia. PMID- 10833905 TI - Ductal sebaceoma (sebomatricoma). AB - We present a case of an unusual tumour arising in the forehead of a 52-year-old female. The tumour, diagnosed as sebaceoma (or sebomatricoma), showed predominantly differentiation towards ductal protion of the sebaceous gland. It was marked in tissue sections as areas of "poroid" cells and the tumour required differentiation from poroma. Focal areas of necrosis en masse in the present tumour contributed further difficulties in to the differential diagnosis. As single multivacuolar sebocytes were found and no cuticular cells could be identified, the tumour was diagnosed as ductal sebaceoma (sebomatricoma). The aspects of morphological distinction between sebomatricomas and poromas are presented with its clinical implications. PMID- 10833906 TI - [Sleep problems]. PMID- 10833907 TI - [Fluid therapy in hypovolemia]. PMID- 10833908 TI - [Children with attention disorders and/or hyperactivity]. PMID- 10833909 TI - [Management of chronic obstructive lung disease]. PMID- 10833910 TI - [An evaluation of a rehabilitation program for patients with chronic obstructive lung disease]. AB - Rehabilitation programmes for patients suffering from chronic obstructive pulmonary diseases are established in many countries. In this paper a new rehabilitation programme at Ulleval University Hospital is described and evaluated. We compared patients' (N = 80) use of hospital inpatient services the year before and the year after attending a rehabilitation course. We also compared patients' self-assessed health status before and six and 12 months after rehabilitation using the St. George's Respiratory Questionnaire and the Hospital Anxiety and Depression scale. The percentage of patients needing hospitalization per year was reduced from 45% before the programme to 22% afterwards. There was no change in the total number of hospitalizations or in the number of days spent in hospital. Self-assessed health status had improved. The results are in accordance with reports from similar rehabilitation programmes in other countries. PMID- 10833911 TI - [Oxygen therapy of patients with chronic obstructive pulmonary disease reduces the development of pulmonary hypertension]. AB - The development of secondary pulmonary hypertension in patients with chronic obstructive pulmonary disease worsens their prognosis; oxygen therapy seems to improve prognosis in these patients. We measured the effect of oxygen therapy on mean pulmonary artery pressure in four women and four men aged 64 to 82 years (average 73 years) with chronic obstructive pulmonary disease. A right cardiac catheterisation was performed before and on average 11 months (8-16 months) after start of oxygen therapy. The mean pulmonary artery pressure decreased on average 5 mm Hg, from 24 to 19 mm Hg. As prognosis seems to be associated with pulmonary artery pressure, more patients with chronic obstructive pulmonary disease should be evaluated for oxygen therapy. PMID- 10833912 TI - [Endoscopic retrograde cholangiopancreatography--a 4-year retrospective study]. AB - Endoscopic retrograde cholangiopancreatography (ERCP) may lead to serious complications. Recently, magnetic resonance cholangiopancreatography (MRCP) has been introduced as a diagnostic alternative to ERCP. This study was initiated to document the diagnostic and therapeutic capabilities of ERCP, enabling us to compare the two techniques. Results of 567 ERCP procedures in 371 patients were reviewed. Bile duct stones were the most frequent indication for the procedure (66%). Normal duct systems (37%) and common bile duct stones (35%) were the most frequent findings. Stone extraction was performed in 97 patients. In 18 patients minor stones were left behind and in six patients open choledocholithotomy was performed. Procedure related mortality was 0.3% and 0.8% in the diagnostic and therapeutic group respectively. Five patients developed serious pancreatitis, and duodenal perforation complicated two procedures. 56% of the procedures were diagnostic and could probably have been replaced by MRCP if this technique had been available during the study period. PMID- 10833913 TI - [Thrombosis prophylaxis in orthopedic surgery]. AB - At Trondheim University Hospital, prophylaxis against thromboembolism after orthopaedic surgery was changed from dextran 70 to low molecular weight heparin in 1992. We wanted to assess whether the frequency of thromboembolic complications has fallen after this change of procedure and to search for any indication that prophylaxis should be prolonged from two to six weeks. Our register showed 97 cases (0.76%) with thromboembolism during the first 12 weeks after 12,711 operations during the dextran 70 period; 99 cases (0.54%) after 18,368 operations during the low molecular-weight heparin period (p < 0.05). A more thorough search identified 121 (0.66%) thromboembolic complications during the low molecular-weight heparin period; 2.2% among patients with total hip arthroplasty and 1.5% among patients with hip fractures. In these two groups, 19 (0.71%) thromboembolic complications were diagnosed during the third to sixth postoperative week. If all these complications had been avoided by prolonging prophylaxis to six weeks, each would have cost around NOK 200,000. Among patients with a diagnosed thromboembolic complication one hip fracture patient and no hip arthroplasty patients died during the third to sixth postoperative weeks. We conclude that there has been a significant fall in thromboembolism following the change from dextran 70 to low molecular-weight heparin. The frequency of thromboembolism and fatal lung embolus is so low during the third to sixth postoperative weeks that prophylaxis beyond two weeks is unwarranted. PMID- 10833914 TI - [Cystic fibrosis in the Western health region]. AB - 78 patients with cystic fibrosis have been diagnosed in western Norway (Health Region 3) since 1960. 31 were born before 1971 (40%). 44 patients were born in the county of Hordaland (62%), including the city of Bergen where the western Cystic Fibrosis Centre is located. 24 (31%) patients were born in Rogaland, and five (7%) in Sogn & Fjordane. 12 patients have died, 9 of them born before 1971 (survival in this cohort of patients is 71%). During two decades from 1971 to 1990, 3 out of 33 patients have died (survival 91%). All patients born after 1990 are alive. The median age at death was 18.5 years. Among patients alive in 1999, 32 (41.5%) are more than 20 years and 19 (24.7%) are more than 30 years old. Median age among all patients is 19.5 years (range 3-51). Chronic lung infection with Pseudomonas aeruginosa has been diagnosed in 20.8% of all patients, which is lower than the average of Norwegian cystic fibrosis patients (35%). Survival among cystic fibrosis patients in Western Norway is good compared to data from other countries. It seems realistic to expect that our patients should achieve a median survival to more than 30 years of age. PMID- 10833915 TI - [Giant urinary bladder stone]. AB - We report a 660 gram stone in the urinary bladder of an otherwise healthy 69 year old male. The stone gave rise to only minor subjective symptoms, but urograms revealed dilatation of both the ureters. The stone was removed by open cystolithotomy. Giant bladder stones are rare in modern urologic practice. In earlier centuries, however, bladder stone formation was a common urinary tract disease with need for surgery. A brief survey of historical lithotomy methods is given. PMID- 10833916 TI - [Diet in Parkinson disease]. AB - The interest in a protein redistribution diet, also called daytime protein restriction diet, has increased among patients with Parkinson's disease. Since certain amino acids compete with L-dopa in the intestine and at the blood-brain barrier, daytime protein restriction may improve fluctuations in motor ability. However, this diet can contribute to weight loss, nutrient deficiencies and cause cognitive disabilities if the diet is not correctly observed. Further studies are needed to clarify how medication with L-dopa in combination with different diets (relative contributions of protein, fat and carbohydrate) may affect motor fluctuations, nutritional status and cognitive ability. PMID- 10833917 TI - [Chronic sleep problems--possible to treat?]. AB - Epidemiological studies show that approximately 10% of the population suffer from chronic insomnia. Insomnia can be classified as primary (without any obvious cause) or secondary to other disorders. Non-pharmacological treatment (i.e. sleep restriction, stimulus control treatment) is recommended in chronic primary insomnia, but this kind of treatment is unknown to most Norwegian health-care providers. This paper presents data from such treatment at a sleep disorders centre in Bergen, Norway. Twenty-two patients (10 men, 12 women; mean age 40 years) with chronic primary insomnia were included in the study. They had on average had insomnia for nine years. They went through a structured behavioural treatment regime with sleep restriction and stimulus control as the main components. Median number of consultations was five. The patients completed sleep diaries every week, from two weeks before the treatment started. Weekly averages obtained from the sleep diaries before and after behavioural treatment were statistically evaluated. Total wake time (summation of sleep-onset latency, wake after sleep onset and early morning awakening) was reduced by two hours. Sleep efficiency (total sleep time divided by time in bed) increased significantly from 65% to 86%. Total sleep time and subjective sleep quality also improved. The use of hypnotic medication was significantly reduced. The results indicate that this non-pharmacological treatment is effective in chronic primary insomnia, also in a Norwegian population. There was no control group in this study, and it should be interpreted with caution. However, randomized controlled trials from other countries document the efficacy of sleep restriction and stimulus control treatment in chronic insomnia. PMID- 10833918 TI - [Hyperactive children meeting the health service system]. AB - Parents of children with hyperkinetic disorders (MBD, AD/HD) often criticize the health care system for being reluctant in recognizing the children's problematic behaviour and reluctant in examining and treating the problems at an early stage. They are also frustrated by insufficient information, inadequate treatment and too little practical help. In collaboration with the Norwegian MBD Association, an anonymous questionnaire was sent to members of the association with children born in 1983 or later. 192 of 350 questionnaires were returned and are included in the analysis. Parents had started being concerned about their child's development very early. When the child was one year old, more than 1/3 of the parents were concerned, at the age of two almost 2/3 of the parents were uneasy or alarmed. However, about 50% of the children were older than three years before the health care system recognized the problems, the average age for receiving a diagnosis was between five and six. Most parents felt there was a lack of sufficient information to the family from professionals, especially from medical doctors and psychologists. The support and information given by the MBD Association were highly appreciated. The study documents that parents need help and support concerning their difficult and demanding baby or toddler long before a diagnosis can be given. The importance of early intervention is strongly emphasised. This represents a challenge for the primary health care system and for the specialist services for children. PMID- 10833919 TI - [DNA analyses in diagnostic pathology]. AB - Since the publication of the polymerase chain reaction (PCR) technique in 1985, there has been an explosion of reports on its use in medicine and science. In this overview the use and limitations of PCR in diagnostic pathology are discussed. As biopsies get smaller, molecular technology will play an ever greater role in improving diagnostic accuracy. PCR is useful in tumour characterisation by assessment of clonality and detection of specific chromosomal rearrangements. This way better targeting of therapeutic intervention is possible. The value of PCR in the diagnosis of cancer, HLA typing and the detection of microorganisms is illustrated by some case reports. PMID- 10833920 TI - [Diagnostic and therapeutic localized breast biopsy]. AB - Preoperative localized mammary biopsy has been used as a diagnostic tool since before 1983 in the Breast Clinic, Ulleval Hospital. The great majority of procedures have been performed in the outpatient clinic with use of local anesthesia. Since 1988 all activity has been registered prospectively in a database. We discuss the use of the procedure and the results obtained during the period 1983-95. In the later part of the observed period, the use of the procedure as a diagnostic tool has been less frequent, but it has been increasingly used as a tool to identify the pathologic process when performing breast-conserving surgery. Surveillance of the malign-benign ratio is an important parameter in the quality assurance of the procedure. The malign-benign ratio has been greater than 0.3 during the period of observation. Continuous registration of procedure-related parameters is necessary for documentation of the results. PMID- 10833922 TI - [Can the international relief work be better?]. PMID- 10833921 TI - [Microdissection--an important tool in molecular biology]. AB - Our knowledge of cell biology is increasing rapidly through the introduction of new molecular biology techniques. Pathological processes always involve a number of cell types; obviously, there is a critical need to know exactly what is being examined. This can be achieved by applying in situ techniques (immunohistochemistry, in situ hybridization, and in situ PCR) on the light microscopical and electron microscopical levels. Microdissection of cells/tissues is an alternative approach. This can be done manually or with the help of a laser, which allows the dissection of single cells. The methods can be used on frozen tissues as well as on paraffin embedded material. Although the ensuing DNA material is not always optimal for further analysis, it is possible to apply special techniques to approve the situation, such as whole genome amplification PCR. Microdissection of cell clones allows us to analyse interactions between tumour cells and stromal cells. Microdissection is based on a simple principle and has proved to be very important in molecular biology analysis of histological material. PMID- 10833923 TI - [Gastroesophageal reflux in children]. PMID- 10833924 TI - [Peroral anticoagulant treatment--are the INR values reliable?]. PMID- 10833925 TI - [Continuing education in anesthesiology]. PMID- 10833926 TI - [Distribution of research resources is problematic]. PMID- 10833927 TI - [Research challenge of general practice]. PMID- 10833928 TI - [Geriatric psychiatry--an example to be followed]. PMID- 10833929 TI - [Treatment of depression in general practice]. PMID- 10833930 TI - [Six years of experiences from a department of geriatric psychiatry]. AB - After a project period from 1994 to 1997, all hospital-based psychogeriatric services in the Norwegian county of Telemark have been located to one department in the central hospital. The department has three differentiated bed units and one out-patient unit. This article describes the psychogeriatric department before and after the project, and includes an evaluation of the quality of service. The changes following the reorganization have been evaluated with an internal analysis, questionnaires to primary health care personnel and an examination of medical and psychiatric records. In 1995 and 1997, questionnaires were sent to general practitioners and district nurses in Telemark County. An independent consultant examined 40 medical and psychiatric records in 1995 and 1997, evaluating them according to selected quality standards. The response rates among doctors were 59% in 1995 and 48% in 1997. The responses show that primary health care doctors and nurses make use of the department's services and that they are increasingly satisfied with the quality of these services. The examination of medical and psychiatric records shows insufficient documentation of psychiatric symptoms and activities of daily life in 1995, but significant improvements by 1997. There are drawbacks to the evaluation method employed. However, we find it warranted to conclude that the reorganization has been functional and that the quality of services has improved. PMID- 10833931 TI - [Endoscopic treatment of vesicoureteral reflux in children]. AB - In the five-year period 1990-95, 102 patients, 78 girls and 24 boys, aged ten weeks to 14 years were treated for grade III-V vesicoureteral reflux by the same surgeon using subureteric teflon injection--the STING procedure. Both primary, secondary and complex reflux were included. The reflux had disappeared in 90 of 133 refluxing ureters, the grade of reflux had diminished in 17 and were unchanged in 26 at the control three months after treatment. After re-STING in 22 reflux ureters, reflux disappeared in 13, was reduced in six and unchanged in three. Reappearance of low grade reflux was seen in eight of 88 ureters between the three months and one year control, but only one needed re-STING. Open surgery was performed in 13 patients (18 reflux ureters) because of unchanged reflux after STING, most of them due to complex reflux: duplex ureters, ectopic ureters, ureteroceles and paraureteral diverticula. It is concluded that most patients with vesicoureteral reflux can be successfully treated by using subureteric teflon injection. PMID- 10833932 TI - [Neurological diseases associated with celiac disease]. AB - During the period from May 1997 to October 1998, eight patients with coeliac disease or dermatitis herpetiformis and neurological disorders were admitted to the Department of Neurology, University Hospital of Bergen. The most frequent conditions were polyneuropathy (seven patients) and spinocerebellar ataxia (three patients). Other conditions were lower motor neuron disease, myelopathy, epilepsy and encephalopathy. The patients used various degrees of gluten-free diet at the time of admission. It remains unclear whether there is a shared common pathogenetic mechanism or the neurological disorder is a complication to the coeliac disease. Both vitamin depletion and immunological mechanisms may cause neurological disorder. Neurological manifestations may occur before the gastrointestinal symptoms. With reference to our patients and available literature we discuss prevalence, clinical picture, pathogenesis, treatment and prognosis. Neurologists, gastroenterologists and general practitioners should be aware that coeliac disease can cause neurological diseases, especially polyneuropathy, cerebellar ataxia and encephalopathy. PMID- 10833933 TI - [Radical treatment of bronchial carcinoids with Nd YAG laser]. AB - Bronchial carcinoids belong to the malignant bronchial tumours. Most of them grow only by local expansion, but some may behave in a more aggressive way. During the authors' practice with therapeutic bronchoscopies, we have done Nd YAG laser necrotizing of bronchial carcinoids in order to establish the exact localisation of the tumour basis, or to reduce the tumour for palliative reasons. Four of our six patients have shown no sign of residual tumour growth after five years of observation, and their tumour is considered eradicated. In selected patients it is possible to eradicate bronchial carcinoids with laser necrotization, and thus save them from major surgery. PMID- 10833934 TI - [Tuberculosis of the hand]. AB - Since 1985 there has been an increase in the incidence of skeletal tuberculosis in Denmark. This increase is attributed to a high incidence of tuberculosis among HIV-positive patients, elderly people and the increasing number of immigrants from areas with endemic tuberculosis. We present a case of hand tuberculosis in a young Somali male adult, where the tubercle bacilli find their way to the third metacarpal bone of the left hand, without a history of trauma or previous pulmonary tuberculosis. There was a delay in the treatment due to misdiagnosis of just three weeks. The abscess totally vanished two months after start of treatment. PMID- 10833935 TI - [Treatment of children with asthma--ideals and reality]. AB - Children with bronchial asthma are the largest group of children with chronic disease in general practice. The aim of this study was to obtain information on morbidity, examination, and treatment in children with bronchial asthma. 102 children with current asthma, aged 7-15 years, participated in this study, which used a questionnaire, flow volume spirometry with beta2-agonist reversibility test, and two weeks of registrations of peak flow and symptom score. Most children had mild to moderate asthma; 60% had acceptable asthma control defined by a morbidity index. 64 children used either inhaled steroids or dinatriumcromoglycate prophylactic. 76 children had been examined by a paediatrician. Lung function measurements had been performed in 57 of 69 children with follow-up in general practice during the last 12 months. Allergy tests had been taken of 87 children. With respect to allergen exposure at home, only 30 children reported having an optimal home environment. Despite the extensive follow-up of children with asthma, our study indicates that the asthma control is not up to the standard set by current guidelines. By use of written treatment plans, and increasing knowledge of the disease, triggers and of the treatment of children and their parents, we probably would exploit the potential better. The recommendations and goals of the guidelines should, however, also be evaluated. PMID- 10833936 TI - [Parvovirus B19 infection--an incidious chameleon]. AB - Parvovirus B19 is a common source of infection with a seroprevalence of 60-70 per cent in the adult population. The most common manifestation is erythema infectiosum ("fifth disease"), with exanthem, fever and upper airway symptoms in children. The infection can give rise to a multifaceted clinical picture and is probably underdiagnosed, particularly in risk groups (individuals with haemolytic anaemia or immunosuppression, and fetuses). Serological diagnosis can now be complemented with the demonstration of viral DNA using the PCR (polymerase chain reaction) test in various body fluids, or tissue biopsy. Recent years have witnessed manifest increase in clinical knowledge of parvovirus B19-associated complications, and their diagnosis and treatment. PMID- 10833937 TI - [Classification of idiopathic juvenile arthritis]. AB - We present a review of the criteria for the classification of juvenile arthritides. Historical aspects, the present situation and proposals for new criteria are outlined. The most commonly used classification criteria today are the European juvenile chronic arthritis criteria (JCA), the European League Against Rheumatism (EULAR) criteria, and the American juvenile rheumatoid arthritis (JRA), the American Rheumatoid Association (ARA) criteria. They differ in nomenclature and have different inclusion and exclusion demands. This has made it difficult to compare studies using different criteria. Neither of them can define homogeneous subgroups of disease. The most recent proposal for new classification criteria of juvenile arthritides is that of the International League Against Rheumatism, ILAR. They are primarily designed to define homogeneous subgroups of disease. The goal is also to obtain an international consensus. Advantages and disadvantages are discussed in this article. The criteria have not yet been validated, and should not be used by clinicians until they have been approved by the international scientific society. We also present guidelines recommended for the classification of juvenile arthritis in Norway today. We recommend using the term juvenile arthritis. Disease duration of arthritis must be at least six weeks. A diagnosis of arthritis should not be made on painful and restricted joint movement alone, as is the case in both the EULAR and ARA criteria today, but at least be based on definite swelling of joints verified by either clinical examination and/or by imaging techniques such as ultrasound, X-ray or EMR. PMID- 10833938 TI - [Children's rights in Norwegian hospitals--are children and parents satisfied?]. AB - The rights of children in hospitals in Norway are protected by regulations based on Norwegian legislation. These regulations cover matters such as teaching of hospitalized children, housing of parents, economic rights of parents, and information. We did a questionnaire study among hospitalized children and their parents about their views on how hospitals complied with these regulations. 90 of 131 questionnaires (69%) were returned. There was a considerable gap between what the law demands of continuance in health care during treatment, schooling and information, and what children and parents actually experience. The study shows that the present regulations concerning children's rights in hospitals should be changed. The health authorities should demand documentation from the hospitals as to how the rights of children are ensured in hospitals. PMID- 10833939 TI - [Hand hygiene--is hand disinfection the best solution?]. AB - Nosocomial infections have been recognized as a critical problem in modern medicine and are associated with the quality of health care provided in hospitals. On average, 7-10% of hospitalized patients acquire an infection after hospital admission. A substantial proportion of these result from cross contamination. Transmission of microorganisms by the hands of health care workers is the main cause for spreading. Prevention of bacterial contamination and infection to the patients requires good hand washing or disinfection, and a series of guidelines have been proposed. However, health care workers show poor compliance with such rules. At a time when costs for patient care are increasing and hospitals are threatened by bacterial resistance, prevention of nosocomial infections is an important issue. Thus, many international publications at present discuss the general problems of hand hygiene, still is the most cost effective method in prevention of nosocomial infections. Factors that contribute to poor compliance in hand hygiene have been targeted, and practical solutions in order to improve hand cleansing practices have been suggested. Increased use of the quicker methods of hand disinfection instead of hand washing is one of them. PMID- 10833940 TI - [Early stage ovarian carcinoma--diagnosis and treatment]. AB - The prognosis of patients with early stage ovarian borderline tumour and carcinoma is good. In spite of this most patient management has been unnecessarily heavy. DNA ploidy is an independent and the strongest prognostic predictor of survival with an excellent prognosis in patients with diploid tumours, it should be included in the routine evaluation of patients in order to achieve optimal and individual management. Image cytometry is superior to flow cytometry in predicting prognosis. PMID- 10833941 TI - [Digital image analysis in pathology--exemplified in prostatic cancer]. AB - Digital image analysis is increasingly used in medicine. This paper reviews image analysis methods currently used in pathology. To exemplify established and future use of these methods, we present our own research and available literature on prostate cancer. DNA ploidy examinations mean measuring nuclear DNA content. A review of available literature shows that DNA ploidy is a very good prognostic marker in early and localized prostate cancer. Nevertheless, request for such examinations is sparse, in contrast to what is the case for gynecological cancers. In addition to assessing nuclear DNA content, we have today the means of quantitatively assessing the chromatin organisation. Such nucleotyping and texture analysis of chromatin has been shown to give significant diagnostic and prognostic information both in localized and advanced prostate cancer. DNA ploidy and nucleotyping are methods that analyze features of single cells. New methods for describing tissue architecture in an objective and reproducible way have been developed in our department. Preliminary results demonstrate the ability of these methods to discriminate between groups of prostate cancer patients with good or poor diagnosis. Digital techniques in combination with new methods from molecular biology have a potential for reducing workload, with parallel assessment of a large number of markers on large populations. The combination of traditional, molecular and digital pathology already offers the possibility of improved diagnostic and prognostic activity. However, there is the challenge of actually employing these methods. DNA ploidy for localized prostate cancer is only one example of a reliable prognostic marker that is not used to its full potential. PMID- 10833942 TI - [Flow cytometry--use in cell biology and benefits in clinical medicine]. AB - Flow cytometry is an automated method for measuring features of single cells in suspension. Many parameters may be measured simultaneously at high speed, objectively and with good statistics. The method has been increasingly used in the context of medical diagnosis and prognosis over the last ten years. Clinical chemical departments are major users of flow cytometry for differential cell counts of leukocyte populations in blood by light scatter measurements. For immunophenotyping of acute leukaemias, flow cytometry is used routinely in all university hospitals in Norway. The method is also used to classify chronic lymphoproliferative disease, and for quantification of CD34-positive stem cells by high-dose chemotherapy with autologous stem cell support. Flow cytometry is routinely used for classifying non-malignant haematological disease like congenital and acquired immunodeficiencies. In diagnostic pathology, DNA flow cytometry is used for analysis of DNA ploidy and S-phase fractions in malignant tumours. DNA aneuploidy is a strong prognostic factor in some childhood tumours and in gynaecological malignant tumours, and flow cytometry is used in tumour biology for measurements of growth and apoptotic cell death. Furthermore, the method is an important research tool in many areas of biomedical research for detection of structural and functional cellular features. PMID- 10833943 TI - [Computer-based morphometric image analysis of endometrial hyperplasia]. AB - Former studies have shown that only 20% of endometrial hyperplasias progress to carcinoma. Objective criteria for predicting the outcome of hyperplasias have been lacking. Because hysterectomy has been the therapy of choice to avoid malignant development, overtreatment of these patients is still a clinical problem. A computer-based morphometric image analysis system has proved to be a considerable improvement for the pathologist in assessing which patients will develop cancer. The system is based on ten nuclear and twelve glandular architectural features. The three parameters with the best statistical power of discriminating between patients developing malignancy and those remaining healthy were combined into a morphometric classification rule (D-score or discriminating score). We give a brief description of this method and its background. PMID- 10833944 TI - [Alternatives of alternative medicine]. PMID- 10833945 TI - [Abortion--one more comment]. PMID- 10833946 TI - [Thrombocytosis and hyperkalemia]. PMID- 10833947 TI - [Admissions to emergency psychiatric departments]. PMID- 10833948 TI - [General practice at the turn of century]. PMID- 10833949 TI - [Methadone therapy in opiate dependency]. PMID- 10833950 TI - [After chromosome 22]. PMID- 10833951 TI - [Malignant melanomas and young men]. PMID- 10833952 TI - [Treatment of congenital heart defects with catheter]. PMID- 10833953 TI - [Induced abortion because of fetal abnormality]. PMID- 10833954 TI - [Infection and pregnancy outcome, use and linkage of data]. PMID- 10833955 TI - [Induced abortion because of fetal abnormality in Norway, 1996-7]. AB - BACKGROUND: The objective of this study was to estimate the national and regional rates of induced abortions after the 12th gestational week attributed to fetal maldevelopment in Norway during 1996 and 1997. MATERIALS AND METHODS: In 1998, the National Board of Health requested from all hospitals copies of relevant documents in the medical records of women who had applied for induced abortion after 12th gestational week. RESULTS: There were 303 women in 1996 and 1997 who applied for induced abortion because of diagnosed fetal maldevelopment. For all of these women the application was accepted. One woman did not have an abortion. The national rate of induced abortion attributed to fetal maldevelopment was 2.5 per 1000 births. There were regional differences in the rate of abortion. In 53% of all applications for induced abortion the fetal diagnosis was not reported. Among the reported diagnoses, 28% were chromosomal abnormalities. INTERPRETATION: There is a need for better knowledge of factors influencing the number of induced abortions attributed to fetal malformations. We also need a better understanding of the impact of such abortions on the observed perinatal mortality and occurrence of birth defects. PMID- 10833956 TI - [Hepatitis C virus among pregnant women in Norway--occurrence of antibodies and pregnancy outcome]. AB - BACKGROUND: Transmission of hepatitis C virus from mother to child is well documented. The prevalence of antibodies against hepatitis C virus among pregnant women in Norway is however, not known. The aim of this study was to estimate the maternal prevalence of antibodies against hepatitis C virus and to study the association between presence of antibodies and fetal death. MATERIAL AND METHODS: From a study of 35,940 pregnant women, a random sample of 970 women and all women with fetal death after 16 weeks of gestation (n = 283), were tested for antibodies against hepatitis C virus. RESULTS: 7 out of 970 women in the random sample (0.7%; 0.2-1.3%, 95% confidence interval) had antibodies against hepatitis C virus. The same prevalence (0.7%, 2 out of 283) was found among women with fetal death. INTERPRETATION: The prevalence of antibodies against hepatitis C virus among Norwegian women was unexpectedly high. Further research is necessary to understand the causes and implications of this observation. PMID- 10833957 TI - [Catheter closure of open ductus arteriosus--the first 100 patients]. AB - BACKGROUND: Patent ductus arteriosus is increasingly treated with catheter based techniques. We present our results as by the end of 1998 of the first 100 patients given such treatment. MATERIAL AND METHODS: The 100 patients between 0.6 and 31.4 years were initially treated with one of these devices: The Rashkind umbrella (60 patients), Cook PDA coils (31 patients) or Amplatzer ductal occluders (ten patients). Seven umbrella patients were treated twice, one with an additional umbrella, six with coils; two coil patients twice, one with another coil and one--after embolisation--with a peg. RESULTS: The overall complete closure rate for all groups was 90% after nine patients had been treated a second time. The primary complete closure rate in pegs was 100%, after reintervention 85% in umbrella and 97% in coil patients. Neither umbrellas nor pegs have embolised. One coil embolised in the course of implantation of 40 coils (2.5%). No other complication has occurred. INTERPRETATION: The closure of the arterial duct with catheter techniques compares favorably with surgery and is now established as the method of choice. Following the investigational introduction of pegs there is no longer an upper limit in the size of ducts suitable for such treatment. PMID- 10833958 TI - [Active uremia therapy in autosomal dominant polycystic kidney disease]. AB - BACKGROUND: Autosomal dominant polycystic kidney disease is the most frequent inheritable kidney disease. Epidemiological studies have not been performed in Norway. MATERIAL AND METHODS: Based on the Norwegian Renal Registry, we retrospectively studied 375 patients diagnosed with this disease taken into renal replacement therapy between 1980 and 1997. They were compared with patients with glomerulonephritis. RESULTS: The average age for autosomal dominant polycystic kidney disease patients at start of renal replacement therapy was 55.2 years with a significant difference (p < 0.003) between men (53.7 years) and women (57.1 years). 295 patients (78.7%) were transplanted while 80 (21.3%) were not transplanted. From start of renal replacement therapy autosomal dominant polycystic kidney disease patients had a significantly higher (p = 0.0002) five year survival rate (70%) compared with glomerulonephritis patients (60%). One and five year patient survival rates were significantly higher (p = 0.0006) for living donor recipients (95% and 90%) compared with necro-kidney recipients (90% and 75%). One year graft survival rates for kidneys from living donors was 90% and 80% for necro-kidneys, after five years the graft survival rate was 80% and 65% (p = 0.0026). After an average of five years, 227 (60.5%) patients are alive, 197 with functioning kidney grafts (87%) while 30 (13%) are on dialysis. INTERPRETATION: Male autosomal dominant polycystic kidney disease patients start renal replacement therapy earlier than female patients. Autosomal dominant polycystic kidney disease patients do have better patient survival in renal replacement therapy than patients with glomerulonephritis. PMID- 10833959 TI - [Hyperbaric oxygen treatment for radiation reactions]. AB - BACKGROUND: A national hyperbaric centre was established in 1994 at Haukeland Hospital with responsibility for all hyperbaric oxygen (HBO) treatment in Norway. In hypoxic tissues with symptomatic radiation reactions, hyperbaric oxygen induces the formation of collagen and angiogenesis resulting in permanently improved local microcirculation. MATERIAL AND METHODS: 234 patients received elective HBO treatment at Haukeland Hospital in 1997 with a total of 4,048 treatments. All 47 patients treated for radiation reactions in the pelvic area in 1997 received a questionnaire 3-15 months after HBO therapy; 81% responded. RESULTS: Rectal bleeding and haematuria were reported as much improved in 61% and 55% respectively, while bladder incontinence was much improved in 46%. INTERPRETATION: This treatment modality may be an alternative in symptomatic radiation reactions of the urinary bladder and the bowel when conventional treatment has given unsatisfactory results. PMID- 10833960 TI - [Diagnostic delay in malignant melanoma]. AB - BACKGROUND: Malignant melanoma accounts for 1 to 3% of all cancers and has been the most rapidly increasing type of cancer during the last decades. Early diagnosis and treatment favours a good prognosis. We wanted to investigate delays in the diagnostic process and patients' knowledge concerning malignant melanoma. MATERIAL AND METHODS: 457 patients with primary cutaneous malignant melanoma received a questionnaire through their physician; 352 (77%) returned the questionnaire. RESULTS: Median patient delay, defined as time between the patient's first observation of changes in a naevus and the first medical consultation, was eight weeks. Younger men had the longest patient delays. Median professional delay, defined as time from the first medical consultation to the time of diagnosis, was one week. 60% of the patients observing changes in a naevus did not initially seek medical advice, as they did not believe the changes were significant. In 65% of the cases, the patients themselves initiated the consultation. Television and other media were principal sources of information. INTERPRETATION: Public campaigns should be designed to reach younger men in particular and focus on self-examination of naevi and immediate contact with a physician when a suspicious lesion is discovered. PMID- 10833961 TI - [Outbreak of multiresistant Acinetobacter baumannii infection]. AB - BACKGROUND: Nosocomial infections caused by multiresistant gram-negative bacteria represent an increasing problem, especially among intensive care patients. A serious outbreak of infection caused by multi-resistant Acinetobacter baumannii occurred in four burn patients. Acinetobacter is a gram-negative coccibacilli which is widespread in nature, and has been reported as an increasing problem in critically ill patients. MATERIALS AND METHODS: The outbreak strain was introduced from Alicante, Spain, by a transferred patient. This strain was resistant to all commonly available systemic antibiotics (including the karbapenems and all aminoglycosides), and sensitive only to polymyxin B. Two patients were critically ill, one of them died in septic shock. RESULTS: The ward was closed for admission of new patients and hygiene precautions were strengthened. Extensive testing of staff and equipment revealed multi-resistant A baumannii on a shower trolley shared by several patients. The outbreak strain was also identified by restriction endonuclease analysis. The patients were kept strictly isolated until their burn wounds were sufficiently healed to allow them to be discharged to their homes. INTERPRETATION: Following discharge of the last patient and extensive cleaning and disinfection of the entire ward, the particularly resistant strain has not reoccurred. Still, this experience may warrant screening for multiresistant gram-negative rods in patients transferred from regions where broad resistance to antibiotics is a common problem. PMID- 10833962 TI - [Tetany--a first symptom of celiac disease]. AB - BACKGROUND: Seizures in children are usually caused by fever or epilepsy, though they may also be caused by electrolyte, vitamin or mineral disturbances. We describe a case of hypocalcaemic tetany with hypovitaminosis D. MATERIAL AND METHODS: The patient was a previously healthy eight months old girl of Indian origin. RESULTS: Investigations revealed that she suffered from coeliac disease. INTERPRETATION: Coeliac disease is a well-known disorder, characterised by enteropathy and malabsorption causing symptoms such as vomiting, diarrhoea and failure to thrive. Enteropathy and specific malnutrition may occur in the total absence of gastrointestinal symptoms. PMID- 10833963 TI - [Felty's syndrome]. AB - BACKGROUND: Felty's syndrome is a complication of rheumatoid arthritis whereby patients develop neutropenia of varying severity and splenomegaly. The major sources of morbidity and mortality are recurrent local and systemic infections, although some patients remain asymptomatic. MATERIAL AND METHODS: In this paper two patients with Felty's syndrome are presented. RESULTS: One patient had recurrent infections. Clinical manifestations, laboratory features and different modalities of treatment are reviewed. INTERPRETATION: Splenectomy has long been standard therapy, but disease modifying antirheumatic drugs (such as gold salts and methotrexate) and colony stimulating factors should also be considered in Felty's neutropenia complicated with infections. PMID- 10833964 TI - [Acetazolamide--are there reasons for its revival in antiepileptic treatment?]. AB - BACKGROUND: Since 1950 the carbonic anhydrase inhibitor acetazolamide has been used as an antiepileptic drug. Because of its tolerance developing properties, acetazolamide has probably been bypassed by the new antiepileptic drugs on the market. However, when taken for 14 day periods with one week's stop in between, acetazolamide is still of value in the treatment of epilepsy. MATERIAL AND METHODS: The paper discusses acetazolamide and its present use in antiepileptic treatment in the light of existing internal control and quality assurance requirements in the medical services. RESULTS: Irrespective og type of seizure, about 90% of patients have an initial reliable effect from acetazolamide, though it is uncertain how long the effect will last after repeated periods of use. As an additional drug, acetazolamide may be particularly well suited for women with menses-related seizures. INTERPRETATION: Acetazolamide may be the drug of choice when drug interaction is a problem, when rapid onset of effect is wanted, or when an additional drug is needed for a short period of time only. PMID- 10833965 TI - [Hepatitis C virus infection among pregnant women and their children in Norway]. AB - Estimates indicate the births of 6 to 78 children vertically infected with hepatitis C virus infection each year in Norway. There is insufficient knowledge of the magnitude of this health problem and the National Institute of Public Health commissioned the authors to approach issues relating to vertical transmission of hepatitis C virus (HCV) infection in Norway. The risk of vertical transmission of HCV appears to be associated with the titre of the maternal viral load. Vertical transmission from nonviraemic mothers has not been demonstrated. No postexposure prophylaxis exists. There is a lack of association between vertical HCV transmission and delivery mode and no association with breast feeding. Universal screening for HCV infection among pregnant women is not recommended. Children born to women known to be HCV-positive should be followed up with antibody and polymerase chain reaction investigations in order to clarify their HCV status. More studies of HCV infection among pregnant women and their children in Norway are needed. PMID- 10833966 TI - [Magnetic resonance tomography in oncology diagnostics]. AB - MR is well suited for imaging in patients with malignant disease. It is the most sensitive and most specific method in the detection of skeletal metastases. It clearly demonstrates the extent of primary bone tumours and also reveals skip metastases and invasion into neighbouring joints, hence it is an important procedure when surgery of a bone tumour is planned. In case of a soft tissue tumour, MR is the preferable imaging modality because it demonstrates the anatomy and reveals the different tissue constituents. MR is the best method to show tumour manifestations in the central nervous system, and it supplements cerebrospinal fluid examination in the detection of meningeal metastasis. MR is the method of choice if there is a cord compression. MR imaging is important in gynecologic oncology. Cost savings have been reported due to reduced use of other diagnostic tests and expensive surgical procedures. New and faster techniques allow examination of the total body in less than 45 minutes. We expect that MR imaging will be increasingly used in the future in patients with malignant disease. PMID- 10833967 TI - [Magnetic resonance tomography in skeletal and soft tissue trauma]. AB - MRI has revolutionized the diagnostic yield in musculo-skeletal trauma. Studies have documented that MRI can be an accurate, cost-effective means of assessing injuries in the knee, the foot and the ankle, and it may also be cost-effective in other anatomic locations. MRI may have a significant impact on decision-making in relation to these patients and on the follow-up. The patient does not need to be moved for evaluation in all the anatomical planes. Each study can also be post processed if necessary. MRI may be used in patients with fractures for evaluation of complications. The fracture lines as well as accompanying soft tissue damage are well documented. PMID- 10833968 TI - [Should pregnant women be tested for antibodies against hepatitis C virus?]. PMID- 10833969 TI - [What is wrong with selective abortion?]. PMID- 10833970 TI - [Ultrasound assessment of gestational age]. PMID- 10833971 TI - [Abortion]. PMID- 10833972 TI - [Codeine is a prodrug--the active agent is morphine]. PMID- 10833974 TI - Identifying systematic reviews: key resources. PMID- 10833973 TI - [Sick leave]. PMID- 10833975 TI - Drug dependence in a journal club. PMID- 10833976 TI - Drug dependence in a journal club. PMID- 10833977 TI - Drug dependence in a journal club. PMID- 10833978 TI - Drug dependence in a journal club. PMID- 10833979 TI - Fine needle aspiration cytology in extramedullary plasmacytoma. AB - OBJECTIVE: Extramedullary plasmacytoma is a rare plasma cell neoplasm. It can occur as the sole manifestation of plasma cell neoplasm, as a metastasis from another extramedullary plasmacytoma, as a solitary plasmacytoma of the bone or as a consequence of multiple myeloma. These plasma cell tumors can occur anywhere and have to be differentiated from other neoplasms, infectious processes and chloroma. STUDY DESIGN: We report the findings of fine needle aspiration cytology (FNAC) in 18 patients with extramedullary plasmacytoma. In six patients extramedullary plasmacytoma was the initial presentation of plasma cell neoplasm. In the remaining 12 patients the tumors occurred under or after treatment of plasma cell disease. RESULTS: Eleven lesions were located in the skin, seven in the lymph nodes, one in the liver and another in the spleen. Two patients with known diagnoses of plasma cell disease were thought, before FNAC, to have an infection, and two had a histologic diagnosis of non-Hodgkin's lymphoma. In 13 of 18 patients, cytologic smears showed anaplastic plasma cells. CONCLUSION: FNAC is a front-line investigative procedure in diagnosing extramedullary plasmacytoma. PMID- 10833980 TI - Tuberculosis of the breast. A cytomorphologic study. AB - OBJECTIVE: Extrapulmonary tuberculosis occurring in the breast is rare despite the fact that 1-2 billion people worldwide suffer from tuberculosis. The aim of this study was to examine the cytomorphology of breast tuberculosis (breast TB) and to review the literature. STUDY DESIGN: Old records from the Cytopathology Laboratory, All India Institute of Medical Sciences, were reviewed from January 1980 to December 1998. Cases of breast TB where a cytologic diagnosis was rendered or a histologic diagnosis with prior fine needle aspiration cytology (FNAC) was available were selected. These slides were reviewed for determining the cytologic findings. RESULTS: One hundred sixty cases of breast TB were included in the study. Six males and 154 females with a clinical suspicion of carcinoma had undergone FNA that was reported as TB. The majority of the patients (111) were in the reproductive age group, 21-40 years. Of the 160 cases, 118 (73.75%) had cytomorphology diagnostic of tuberculosis--epithelioid cell granulomas with caseous necrosis. Eleven of the remaining 42 cases were positive for acid-fast bacilli (AFB) on Ziehl-Neelsen (ZN) staining, while 31 cases were confirmed to be tubercular on histology. ZN staining was done in 44 cases, and AFB were demonstrated in only 38.6% of cases. CONCLUSION: Up to 73% of breast TB can be confidently diagnosed when both epithelioid cell granulomas and necrosis are present. Also, the possibility that a woman in the reproductive age group who presents with a palpable lump in the breast may have tuberculosis must be kept in mind, especially as the incidence of breast TB may increase in the future with the global spread of AIDS. PMID- 10833981 TI - Acid-fast bacilli in fine needle aspiration smears from tuberculous lymph nodes. Where to look for them. AB - OBJECTIVE: To analyze where acid-fast bacilli (AFB) are most often seen in smears prepared from tuberculous lymph nodes. STUDY DESIGN: Patients referred for fine needle aspiration cytology for evaluation of lymphadenopathy between March 1994 and June 1998 were analyzed. Only those cases clinically and therapeutically proven to be tuberculous were included in the study. RESULTS: Of 783 cases analyzed, 213 (27.2%) were tuberculous. Aspirates obtained were of three types: blood-mixed particles, caseous material and pus. Five cytologic pictures were seen: epithelioid cell granulomas alone or with coexistent necrosis, AFB or both, and necrosis with AFB. AFB were most often seen in purulent aspirates, followed by caseous and least often in blood-mixed particles. Granulomas were most often seen when the aspirate was blood-mixed particles, followed by caseous and, least often, pus. CONCLUSION: AFB detection should be carried out on all suspected tuberculous patients. The relationship between the presence of granuloma and of AFB is inverse. The chance of finding AFB is highest in patients presenting with a cold abscess and yielding pus on aspiration followed by patients who yield caseous material on aspiration. PMID- 10833982 TI - Cyclin E expression and early cervical neoplasia in ThinPrep specimens. A feasibility study. AB - OBJECTIVE: To investigate cyclin E expression as a possible marker for early cervical neoplasia using ThinPrep gynecologic specimens from premenopausal women. STUDY DESIGN: Archived ThinPrep liquid-based cervical/endocervical specimens (Cytyc Corporation, Boxborough, Massachusetts, U.S.A.) diagnosed as human papillomavirus infection (HPV) (20), atypical squamous cells of undetermined significance (ASCUS) (48) and within normal limits (WNL)/benign cellular changes (BCC) (21) were resampled in duplicate, fixed in 95% ethanol, subjected to immunocytochemical staining with the cyclin E antibody (clone 13A3, Novocastra Laboratories Ltd., Newcastle upon Tyne, U.K.) and HPV antibody (clone K1H8, Dako Corporation, Carpinteria, California, U.S.A.) and the expression scored by two pathologists and correlated with the cytologic diagnosis. A case was scored as positive if it contained > 10 abnormal squamous cells with nuclear immunocytochemical staining. RESULTS: The cylin E antibody assay was positive in 20 (100%) cases cytologically diagnosed as HPV. These cases were also anti-HPV antibody positive. Four cases (19%) cytologically diagnosed as WNL/BCC were cyclin E positive. Of these, two were anti-HPV antibody positive. Thirty-four (73%) cases cytologically diagnosed as ASCUS were positive for the cyclin E assay and for anti-HPV antibody staining. CONCLUSION: Cyclin E expression correlates strongly with morphologic features of HPV in ThinPrep specimens and may serve as a surrogate marker for HPV infection and early cervical preneoplastic lesions. PMID- 10833983 TI - Genital human papillomavirus infection and associated penile intraepithelial neoplasia in males infected with the human immunodeficiency virus. AB - OBJECTIVE: To investigate the association between human papillomavirus (HPV) infections and penile intraepithelial neoplasia (PIN) in genital lesions from human immunodeficiency virus (HIV)+ men. For comparison, we also investigated the same association using specimens from HIV- men. STUDY DESIGN: Imprint smears from penile lesions were obtained from 70 men (mean age, 30 years) who visited a dermatologist. Thirty of them were known to be HIV seropositive. Two study groups were formed: one of 40 HIV- and another of 30 HIV+ males. The smears were examined cytologically for HPV identification or PIN, immunocytochemically for HPV detection and by in situ hybridization for HPV typing. RESULTS: The rates of detecting HPV infection cytologically were higher among men with HIV infection (50%) than among their HIV-seronegative counterparts (30%). There was immunocytochemical evidence of HPV in HIV-infected men in a greater proportion (50%) than in HIV noninfected men (37.5%). By in situ hybridization it was found that there was a higher prevalence of potentially oncogenic HPV (16/18, 31/33/35): 75% in moderate or severe dysplasia (PIN 2 and 3) and 66.6% in HIV+ men as compared with HIV- men (10-16.6%). CONCLUSION: HIV-seropositive males showed an unbalanced distribution of HPV, with a predominance of "high-risk" HPV types. This suggests that immunodepression encourages infection by this oncogenic virus, thereby contributing to the frequency of precancerous lesions in HIV+ men. PMID- 10833984 TI - Combined Pap smear, cervicography and HPV DNA testing in the detection of cervical intraepithelial neoplasia and cancer. AB - OBJECTIVE: The sensitivity of the Pap smear (PAP) continues to be the subject of debate. During the past several years, cervicography (CER) and HPV DNA testing have been suggested as optional tools in the screening of cervical cancer precursors. STUDY DESIGN: The performance characteristics of PAP, CER and HPV DNA testing (hybrid capture test [HCT]) in all potential combinations were evaluated in a series of 1,030 women (aged 16-70, median, 33), subjected to colposcopy (COLPO) as the reference tool. RESULTS: Of the 992 evaluable cases, 402/992 (41%) had positive COLPO (i.e., an abnormal transformation zone). Of them, 298 women underwent directed punch biopsy, while of the COLPO negative patients, 18/93 positive by at least one of the three tests had endocervical curettage. Of the 402 COLPO positive women, 146 (36%) remained negative on all tests, whereas 256 (64%) had at least one positive test. There were 84 cervical intraepithelial neoplasia (CIN) 2 and 3 lesions and 6 invasive carcinomas. Of the former, 10 were detected by PAP alone, 4 by CER alone and 3 by HCT alone. Three of the 6 carcinomas were HCT negative. The predictive value (PPV) of a positive test was 45% for PAP, 51% for CER and 48% for HCT. The combinations of PAP with CER (for PAP negative cases) and PAP with HCT were more sensitive for CIN 2 and 3 (95% and 94%, respectively) as compared with PAP alone but were associated with a significant decrease in specificity (44% and 46% vs. 57%, respectively). However, both combinations retained a PPV (43%) similar to that of PAP alone (45%). CONCLUSION: The potential combinations of PAP with CER and with HCT were more sensitive in detecting CIN 2 and 3 as compared with PAP alone and retained a PPV similar to that of PAP. PMID- 10833985 TI - Endocervical atypical glandular cells of undetermined significance. I. Morphometric and cytologic characterization of cases that "cannot rule out adenocarcinoma in situ". AB - OBJECTIVE: To determine whether evaluating morphologic features through morphometry and cytology can lead to a more-satisfactory characterization of endocervical atypical glandular cells of undetermined significance (AGUS) cases that "cannot rule out adenocarcinoma in situ" (AIS). STUDY DESIGN: Fifty-seven endocervical AGUS cases showing incomplete criteria of AIS were morphometrically compared to five smears with normal endocervical columnar cells (ECC) and to five histologically confirmed endocervical adenocarcinoma cases. For each atypical nucleus, the area and shape were measured. Twenty-five cytologic criteria were used to review the AGUS and neoplastic smears. RESULTS: AGUS nuclei showed an intermediate value in terms of area and shape as compared to the values of normal and neoplastic nuclei. In particular, AGUS nuclear enlargement (136.626 micron 2) was about twice the area of normal nuclei and half the value of the area of neoplastic nuclei (P < .0000). AGUS nuclei also had the greatest variability in size and shape, indicating that anisonucleosis may be a morphologic discriminator of endocervical AGUS. The cytologic features useful in discriminating AGUS from neoplastic smears were: presence of normal ECCs, singly or in sheets (P < .001); absence of necrosis (P < .001); bare atypical cells (P < .001); papillary groups (P < .01); anisonucleosis (P < .05); irregular chromatin distribution (P < .05); and hyperchromasia (P < .01). CONCLUSION: Morphometry and cytology led to a better characterization of endocervical AGUS cases that "cannot rule out AIS." PMID- 10833986 TI - Endocervical atypical glandular cells of undetermined significance. II. Morphometric and cytologic analysis of nuclear features useful in characterizing differently correlated subgroups. AB - OBJECTIVE: To evaluate whether some nuclear features analyzed by morphometry and cytology may be useful in characterizing differently correlated endocervical atypical glandular cells of undetermined significance (AGUS) subgroups. STUDY DESIGN: Fifty-four endocervical AGUS smears were subclassified into four subgroups on the basis of their different correlation: not otherwise specified subgroup (NOSs), tamoxifen (Ts), human papillomavirus infection (HPVs) and laser therapy (LTs). Area and shape of the atypical nuclei detected were morphometrically measured. The smears were then cytologically reviewed, and the shape and grade of expression of hyperchomasia in AGUS nuclei were analyzed. RESULTS: AGUS cases due to T therapy showed the largest nuclear area (148.845 micron 2; P < .0000) and the greatest anisonucleosis objectively measured by morphometry. HPVs had the shape that most differed from perfectly circular (15.341 versus 14.1) and showed the highest grade of expression of nuclear density. LTs and NOSs were less well characterized than the other subgroups. CONCLUSION: Analysis of nuclear features by morphometry and cytology was useful in characterizing the AGUS subgroups Ts and HPVs. PMID- 10833987 TI - Fine needle aspiration cytology of the Ewing's sarcoma family of tumors. AB - OBJECTIVE: To study the cytomorphologic features of the Ewing's sarcoma (ES) family of tumors. STUDY DESIGN: During a period of eight years (1990-1997), 123 soft tissue tumors and 65 bone tumors were evaluated by fine needle aspiration cytology (FNAC); 14 cases were diagnosed as in the ES family of tumors. The ages of the patients ranged from 8 to 30 years. All the cases were histologically confirmed. RESULTS: Of 14 cases of the ES family of tumors, 7 were ES, 3 extraosseous ES (EOE), 2 peripheral primitive neuroectodermal tumor (PPNET) and 2 Askin tumor. Cytologically, smears from all the cases showed round tumor cells with a high nuclear/cytoplasmic ratio. On detailed examination, subtle differentiating features were observed. The cells in ES had finer nuclear chromatin in comparison to those of PPNET and Askin tumor, and punched-out clear cytoplasmic vacuoles were present. PPNET showed nuclear molding, unipolar cytoplasmic tags and Homer-Wright rosettes. Histologically, all cases of ES and EOE and one case of Askin tumor showed periodic acid-Schiff-positive inclusions. CONCLUSION: FNAC features coupled with clinical findings enable a rapid diagnosis of the ES family of tumors, from which treatment modalities can be determined. PMID- 10833988 TI - Fine needle aspiration cytology of soft tissue tumors. AB - OBJECTIVE: To review of the value of fine needle aspiration (FNA) cytology in the diagnosis of soft tissue tumors (STT). STUDY DESIGN: A review of the literature was coupled with the authors' experience with indications, diagnostic specificity and pitfalls; clinical information; and the final cytology report. RESULTS: Over the last few years, FNA has come to be considered a valuable tool in the management of STT in that it affords a specificity of > 90%. FNA is of particular value in any subcutaneous lesion > 5 cm, in all pediatric tumors and whenever direct incision biopsy is particularly contraindicated. Material from aspirates can be used to obtain cytologic smears for conventional staining, special pigment identification, histochemical techniques, cell blocks for paraffin embedding and ancillary techniques (immunocytochemistry, electron microscopy, and densitometric and cytogenetic analyses). The cytologic diagnosis, like its histologic counterpart, should be based on a correct evaluation of clinical data (age, localization, size, effect on bone, nerve and vessel involvement), radiologic information, cytologic findings (architectural pattern, cell and stroma characteristics) and results of special staining techniques. The final cytology report should place the tumor in one of three basic categories: benign, malignant, and inconclusive or undetermined. Wherever possible, a histopathologic diagnosis should also be provided, either based on purely cytologic criteria or aided by ancillary techniques. CONCLUSION: FNA does not present major complications and permits a swift, preliminary diagnosis in a large number of cases. The method is most effective when the aspiration is performed by an experienced pathologist. PMID- 10833989 TI - Computed tomography-guided fine needle aspiration of peripheral lung opacities. An initial diagnostic procedure? AB - OBJECTIVE: To evaluate the results of computed tomography (CT)-guided fine needle aspiration (FNA) cytology following negative fiberoptic bronchoscopy and sputum cytology. STUDY DESIGN: Retrospective study of 64 patients who underwent CT guided needle aspiration of lung opacities over one year. Following a review of the CT studies, patients were selected according to image characteristics of a primary neoplasm and pleural effusion in cases with pleural lesions. The lesions were classified into three categories--intrapulmonary and peripheral pulmonary, pleuropulmonary and pleural--and were localized and aspirated under CT using a fine needle (22-23 gauge) for obtaining cellular material. Lesions diagnosed as benign on FNA cytology were followed by serial CT scans for a period of two years at six-month intervals. RESULTS: Thirty-nine of 64 (61%) lesions were diagnosed as malignant on FNA cytology and 25 of 64 (39%) as benign. There was one false negative case. There were no serious complications from the procedure. CONCLUSION: FNA under CT guidance may be applied as the initial procedure in the diagnosis of peripheral malignant pulmonary lesions, rendering a high diagnostic yield. PMID- 10833990 TI - Testicular fine needle aspiration cytology for the diagnosis of azoospermia and oligospermia. AB - OBJECTIVE: To evaluate qualitative and quantitative cytologic features on testicular fine needle aspiration biopsy in the diagnosis of azoospermia and oligospermia and to correlate cytologic and histologic diagnoses. STUDY DESIGN: In this prospective study, 50 infertile males selected from the infertility clinic of Guru Tegh Bahadur Hospital were studied. Fine needle aspiration cytology (FNAC) smears from both testes of 27 azoospermic and 23 oligospermic patients (sperm count < 10 million per milliliter) were stained with May-Grunwald Giemsa and Papanicolaou stain. Differential counting of 500 spermatogenic cells was done, and the number of Sertoli cells per 500 germ cells was determined for calculating the spermatic index and Sertoli cell index, respectively. FNAC and testicular biopsy were performed under local anesthesia as a minor surgical procedure. RESULTS: Six groups were identified on FNAC smears from azoospermic patients: I. normal spermatogenesis (8), II. hypospermatogenesis (2), III. maturation arrest (2), IV. Sertoli cells only (6), V. atrophic pattern (7), and VI. Leydig cell predominance (2). In oligospermic patients two groups were identified: I. those with normal spermatogenesis (4), and II. those with subnormal spermatogenesis (19). Correlation with histopathologic examination was seen in 81.5% azoospermic and 65.2% oligospermic patients. CONCLUSION: Qualitative and quantitative evaluation of testicular FNAC provides useful information on both azoospermic and oligospermic patients. FNAC performed under local anesthesia is an acceptable outpatient procedure that consistently yields sufficient diagnostic material in all patients. PMID- 10833991 TI - Diagnostic value of hair shafts and squamous cells in peritoneal washing cytology. AB - OBJECTIVE: Little attention has been given to hair shafts and squamous cells in peritoneal fluid. To investigate their diagnostic value in peritoneal washing specimens, we reviewed peritoneal washing cytology preparations from 83 cases of ovarian tumors. STUDY DESIGN: We reviewed peritoneal washing specimens and histologic sections of 86 cases of ovarian tumors and tumorous conditions, including 22 teratomas, 16 serous adenocarcinomas, 10 clear cell adenocarcinomas, 9 endometrioid adenocarcinomas, 5 cases of endometriosis, 4 mucinous adenomas, 3 serous cystadenocarcinomas and 17 other tumors. RESULTS: We observed both squamous cells and hair shafts surrounded by inflammatory cells in 5 of the 22 cases of ovarian teratoma. Rupture of an ovarian teratoma was clinically and histologically found in one of the five cases. Hair shafts were not observed in the other tumors or in nonneoplastic conditions. The diameter of hair shafts in peritoneal washing specimens ranged from 10 to 28.8 microns (average, 16.6), and such hair shafts were present within an ovarian teratoma examined histologically. The diameter of hair shafts from six normal adults who were examined as controls ranged from 61.5 to 118.6 microns (average, 89.4). CONCLUSION: Hair shafts and squamous cells surrounded by inflammatory cells in peritoneal washing specimens are a diagnostic clue to ovarian teratoma and can be observed even when rupture of the tumor is not detected clinically or microscopically. PMID- 10833992 TI - Fine needle aspiration cytology of the breast. Experience at an outpatient breast clinic. AB - OBJECTIVE: To evaluate the accuracy of fine needle aspiration cytology (FNAC) of the breast at our institution and to perform quality assurance. STUDY DESIGN: Two hundred forty-six cases with pathologic confirmation were selected and reviewed. A pathologist performed most of the aspirations at an outpatient breast clinic. We correlated cytologic and histologic findings and evaluated the influence of the size, location, grade, and pathologic subtypes and fibrosis in breast lesions on diagnostic results. RESULTS: The likelihood ratios for malignant, suspicious, atypical, benign and unsatisfactory cytologic diagnoses were 98.71, 5.48, 1.09, 0.07 and 0.55, respectively. The absolute and complete sensitivities for malignant lesions were 64.5% and 90.3%, respectively. The specificity was 71.9%. False negative and positive rates were 4.3% and 0.7%, respectively. The predictive value for a malignant cytologic diagnosis was 98.4%. The rate of unsatisfactory samples was 9.3%. The rate of concordance between cytologic and histologic diagnosis was lower for large and diffusely growing lesions (benign and malignant), for malignancies with abundant fibrosis and of unusual types and for carcinomas of low grade. All axillary and recurrent chest wall lesions were diagnosed cytologically. Cell block sections were useful in a small number of cases. CONCLUSION: Understanding the performance and limitations of FNAC can enhance its value as a diagnostic technique in the management of breast disease. PMID- 10833993 TI - Detection of trisomy 12 by fluorescence in situ hybridization on archival cytopathologic material in chronic lymphocytic leukemia/small lymphocytic lymphoma. AB - OBJECTIVE: To evaluate fluorescence in situ hybridization (FISH) for the detection of trisomy 12 in archival cytologic specimens of chronic lymphocytic leukemia/small lymphocytic lymphoma. STUDY DESIGN: The cytopathology database was searched for all cases of chronic lymphocytic leukemia/small lymphocytic lymphoma. Six cases of chronic lymphocytic leukemia/small lymphocytic lymphoma obtained by fine needle aspiration and one case of small lymphocytic lymphoma with plasmacytoid features were analyzed for trisomy 12 by FISH. These cases had been archived between 1 week to 16 months prior to analysis. RESULTS: We detected trisomy 12 in four of the six cases of small lymphocytic lymphoma/chronic lymphocytic leukemia. The case of small lymphocytic lymphoma with plasmacytoid features was negative for trisomy 12. CONCLUSION: Detection of trisomy 12 by FISH can be effectively performed on routinely prepared, stained and coverslipped archival cytologic material. PMID- 10833994 TI - Malignant teratoma of the thyroid with predominantly neuroepithelial differentiation. Fine needle aspiration cytologic, histologic and immunocytochemical features of a case. AB - BACKGROUND: Teratoma of the thyroid in adults is extremely rare, and most are malignant. Only nine cases have been adequately documented in the English language literature, and there are no reports detailing the fine needle aspiration (FNA) cytologic characteristics. CASE: A 32-year-old female presented with a left-sided nodular thyroid mass with left cervical lymphadenopathy. FNA cytology of the thyroid and lymph nodes was done. The cytologic and immunocytochemical features were that of a small round cell tumor with neuroepithelial (NE) differentiation, metastasizing to the cervical nodes. Microscopic study of the thyroidectomy specimen showed a tumor showing an NE pattern with occasional islands of squamous and cuboidal epithelium, leading to a diagnosis of malignant teratoma. CONCLUSION: Knowledge of FNA cytologic features of rare but highly malignant lesions like thyroid teratomas allow early recognition so that suitable and possibly aggressive treatment protocols can be adopted in the hope of prolonging survival. PMID- 10833995 TI - Renal collecting duct carcinoma. Report of a case with cytologic findings on fine needle aspiration. AB - BACKGROUND: Collecting duct carcinoma (CDC) of the kidney is a rare type of renal cell carcinoma (RCC) of collecting duct origin. Cytologic differentiation of CDC from conventional RCC is important because CDC has a poorer prognosis than the latter. CASE: A 60-year-old male incidentally demonstrated a left renal mass that was hypovascular by angiography. Fine needle aspiration (FNA) revealed numerous clusters of cells arranged in a tubular structure. The cells consisted of highly atypical cells having large nuclei with coarse or vesicular chromatin, prominent nucleoli and lacy or granular cytoplasm. Based on these findings, which were indicative of high grade RCC, he underwent left radical nephrectomy and lymphadenectomy. Histologic and immunohistochemical findings, including anti-high molecular-weight cytokeratin (HMCK) antibody, confirmed the diagnosis of CDC. CONCLUSION: CDC should be added to the differential diagnosis when the result of cytologic examination of a renal mass is suggestive of high grade RCC. These features of FNA smears, together with HMCK immunohistochemistry, can be useful for the cytologic differential diagnosis of renal tumors. PMID- 10833996 TI - Neonatal juvenile xanthogranuloma. Report of a case with fine needle aspiration cytologic findings in a soft tissue mass. AB - BACKGROUND: Juvenile xanthogranuloma is an infrequent, benign histiocytic lesion, the recognition and diagnosis of which by fine needle aspiration biopsy are important for ascertaining whether a case will have a benign course or spontaneous regression. CASE: A case of juvenile xanthogranuloma was located in the upper lip of a newborn male. CONCLUSION: Juvenile xanthogranuloma has characteristic cytologic features that may allow recognition in fine needle aspiration cytology smears. PMID- 10833997 TI - Adenoma malignum. Report of a case with cytologic and colposcopic findings and immunohistochemical staining with antimucin monoclonal antibody HIK-1083. AB - BACKGROUND: Adenoma malignum of the uterine cervix was first described by Gusserow. We report here a case with cytologic, histologic and colposcopic findings and immunohistochemistry for HIK-1083. CASE: A 42-year-old female was noted to have a probable adenoma malignum due to the detection of atypical cells classified as V. On colposcopy, comma-shaped, atypical vessels spread over the entire cervical area. Histologic findings were characteristic of tumor invasion beyond the layer of cervical glandular ducts. Immunohistochemical detection of CEA was negative, but HIK-1083, which recognizes gastric glandular mucous cells, was positive. CONCLUSION: For a definitive diagnosis of adenoma malignum of the cervix, immunohistochemical examination for an appropriate marker, such as HIK 1083, should be added to the routine gynecologic examination, cytologic and histopathologic examination, and colposcopy. PMID- 10833998 TI - Use of immunohistochemistry in fine needle aspiration of thyroid nodules in patients with a history of malignancy. A report of two cases. AB - BACKGROUND: A history of a nonthyroid malignancy may present a diagnostic dilemma in the assessment of fine needle aspiration (FNA) of thyroid nodules. One reported series, on patients with prior malignancies and a thyroid nodule, indicated that in 17% of patients, the thyroid nodule represented metastatic malignancy, 6% were classified as primary thyroid cancers, and the remainder were benign or inconclusive lesions. The resolution of this problem is essential to patient management. CASES: We report two cases in which patients with a history of renal cell carcinoma presented with a thyroid nodule. The first patient was an 80-year-old female whose Papanicolaou-stained FNA demonstrated clusters of round to polygonal cells with round to ovoid, hyperchromatic nuclei and abundant, wispy cytoplasm. The second patient was a 55-year-old female with clusters and single cells with round to oval, eccentric nuclei and copious, granular, gray cytoplasm noted on Papanicolaou-stained material. In each case, the diagnosis was inconclusive on initial review of Papanicolaou-stained slides, and immunohistochemical staining was ordered to better characterize the lesions. Tumor cells from case 1 were positive for cytokeratin cocktail and vimentin and negative for thyroglobulin, epithelial membrane antigen and calcitonin, suggestive of metastatic renal cell carcinoma. In contrast, the tumor cells from case 2 expressed cytokeratin, thyroglobulin and vimentin, consistent with a primary thyroid neoplasm. In each case, the cytologic diagnoses were confirmed in the resected specimens. CONCLUSION: Immunohistochemistry is a helpful adjunct in the evaluation of thyroid nodules in patients with a past history of malignancy. PMID- 10833999 TI - Seminal vesicle epithelium as a potential pitfall in the cytodiagnosis of presacral masses. A report of two cases. AB - BACKGROUND: Whenever abdominoperineal resection is performed because of a rectal adenocarcinoma, the prostate and seminal vesicles may be displaced backward to the presacral space, giving rise to a false radiologic image of a presacral tumor. Due to cytologic atypia associated with the epithelium of seminal vesicles, there is a real possibility, in fine needle aspiration biopsy (FNAB), of erroneously giving a malignant diagnosis. CASES: Two men, aged 53 and 57 years, presented with presacral masses three months and six years, respectively, after abdominoperineal resection for rectal adenocarcinoma. In both cases, FNAB smears showed some groups and single cells with large and irregular nuclei. These cells suggested a recurrence of carcinoma. The presence of cytoplasmic coarse pigment and a background with spermatozoa and blobs of inspissated secretory product were sufficient to determine that these presacral masses represented the seminal vesicles. CONCLUSION: Awareness that seminal vesicles may give rise to a radiologic impression of presacral tumor after abdominoperineal resection of the rectum will avoid unnecessary FNAB and a cytologic false positive diagnosis of colorectal adenocarcinoma. PMID- 10834001 TI - Exfoliative cytology of lymphoepitheliomalike carcinoma of the uterine cervix. A report of two cases. AB - BACKGROUND: Lymphoepitheliomalike carcinomas (LECs) are morphologically similar to undifferentiated nasopharyngeal carcinoma but occur at sites other than the nasopharynx. They rarely occur in the uterine cervix. Sixty-five cases of LEC of the cervix have been published to date, and the pitfalls of histopathologic interpretation have been discussed. This undifferentiated carcinoma with a prominent lymphocytic infiltrate represents a challenge for the pathologist examining a scant cervical biopsy or Pap smear. Distinguishing LEC as a separate entity is important. Despite the fact that the epithelial component is poorly differentiated, this neoplasm is associated with a lower frequency of lymph node metastases, is potentially radiosensitive and has a better prognosis. Although mentioned in passing in several papers, the exfoliative cytology of this cervical neoplasm has not been adequately discussed. We report the cytologic features of LEC in cervical smears obtained from two patients. CASES: The first patient presented with menometrorrhagia and postcoital bleeding. The cervical smear taken at the time of presentation was reported as unsatisfactory for evaluation. ASCUS was diagnosed on a vaginal smear obtained one year earlier. The second patient presented with a complaint of postcoital bleeding. A cervical smear and the cervical biopsy taken at the time of presentation were reported as ASCUS and high grade dysplasia versus carcinoma, respectively. A retrospective review of the cervical smears revealed rare malignant cells occurring singly or in small groups. The tumor cells had a high nuclear/cytoplasmic ratio, irregular nuclear membrane and hyperchromatic nuclei with coarse chromatin and were obscured by heavy inflammation and blood. The background resembled that of a menstrual smear. CONCLUSION: The diagnosis of LEC of the cervix is often made on a loop electrical excision procedure or on a hysterectomy specimen. The presence of heavy inflammation and blood, which can obscure the malignant nature of the cells, presents the cytopathologist with a challenging diagnosis of LEC in cervical smears. In view of the prognostic implications, it is desirable for the pathologist to classify LEC as a distinct entity. PMID- 10834000 TI - Small cell carcinoma of the bladder. Two cases diagnosed by urinary cytology. AB - BACKGROUND: Primary small cell carcinoma (SCC) of the bladder is a rare but important entity. We report two cases of SCC of the bladder diagnosed by urinary cytology. CASES: A 71-year-old male (case 1) and a 79-year-old female (case 2) presented with asymptomatic gross hematuria. Urinary cytology in case 1 showed the presence of a few undifferentiated malignant small cells and many transitional cell carcinoma (TCC) cells with a bloody and necrotic background. The former cells were small and round, with naked, hyperchromatic nuclei and finely granular chromatin. Pathologic diagnosis after total cystectomy was TCC > SCC > adenocarcinoma, T2M0N0. Urinary cytology of case 2 showed the presence of many undifferentiated malignant small cells and many TCC cells with or without squamous metaplasia. Cytologic features of the former cells were almost the same as those in case 1. Moreover, these cells were neuroendocrine marker positive by immunocytochemistry. Pathologic diagnosis after tumor resection was SCC and TCC > squamous cell carcinoma, T1b. CONCLUSION: The prognosis of primary SCC of the bladder is usually poor. Because our cases were found by urinary cytology at a relatively early stage, both have been well, without any evidence of recurrence, 30 and 25 months after surgery even without adjuvant therapy. PMID- 10834002 TI - Myxoid leiomyosarcoma of the uterus. Report of a case with cytologic findings. AB - BACKGROUND: Myxoid leiomyosarcoma is a rare variant of uterine sarcoma, exhibiting malignant biologic behavior despite the absence of cytologic atypia and of significant mitotic activity. CASE: A 20-year-old female was referred with a cystic pelvic mass. At laparotomy, the tumor, weighed 2,200 g and originating in the left lateral uterine wall, was removed. Microscopic examination revealed well-differentiated smooth muscle cells without atypia and with a few mitotic figures in the copious myxoid matrix, suggesting myxoid leiomyosarcoma. Three years following laparotomy, an irregular mass around the uterus was noted on sonographic examination, suggesting local recurrence. Two years and six months later, the second operation was performed, and a locally recurrent, multicystic tumor weighing 3,500 g was excised. The histopathology was similar to that of the primary tumor. Cytologic findings on imprint material from the tumor revealed a few isolated or sheet like small cells consisting of spindle and polygonal cells with round and oval nuclei. Cytologic atypia was also minimal. CONCLUSION: Myxoid leiomyosarcoma should be included in the differential diagnosis of smooth muscle neoplasia. PMID- 10834003 TI - Human papillomavirus infection in atrophic smears. A case report. AB - BACKGROUND: Human papillomavirus (HPV) infection in atrophic smears can be misleading and may produce the diagnosis of cervical intraepithelial neoplasia. CASE: A routine cervical smear in a 62-year-old female revealed an atrophic smear with nuclear changes suggestive of a high grade squamous intraepithelial lesion (HSIL). An estrogen cream for topical vaginal use was prescribed. A new smear was collected seven days later and revealed koilocytosis but no evidence of HSIL. CONCLUSION: Koilocytosis is a cellular finding of mature epithelial cells. The use of estrogen produces maturation of HPV-infected basal cells, allowing a correct diagnosis of this disease in patients with atrophic smears. PMID- 10834004 TI - Fine needle aspiration and touch imprint cytology of a malignant fibrous histiocytoma of the spermatic cord. Case report. AB - BACKGROUND: No cytologic reports on spermatic cord sarcomas have been published. CASE: A 64-year-old man presented with a slowly growing, painless, left spermatic cord enlargement. Fine needle aspiration (FNA) obtained < 1 mL of bloody fluid consisting of solitary, mark-edly anaplastic and pleomorphic tumor giant cells occasionally arranged in small fragments. Rare atypical spindle cells could be observed. Some reactive lymphocytes were observed intermingled with tumor cells. Immunohistochemistry displayed vimentin reactivity and negativity for keratins and leukocytic common antigen. The specimen removed showed a well-circumscribed, 30-mm, yellowish solid tumor. Touch imprints displayed pleomorphic tumor cells showing intense anisonucleosis; a moderate amount of clear, sometimes microvacuolated cytoplasm; and tissue fragments with a storiform pattern. Histologic examination revealed microscopic and immunohistochemical features of malignant fibrous histiocytoma (MFH) arising in soft tissues of the spermatic cord. CONCLUSION: FNA of a spermatic cord lesion may reveal a pleomorphic sarcoma. A pleomorphic appearance together with some spindle elements and compatible immunocytochemistry could help diagnose spermatic cord MFH. This is one of the few reports dealing with FNA cytology of paratesticular tumors and the first report, to the best of our knowledge, showing the cytologic characteristics of a case of spermatic cord MFH. PMID- 10834005 TI - Liesegang rings in cytologic samples accompanied by calcium oxalate-like crystals. A report of three cases. AB - BACKGROUND: The presence of Liesegang rings (LRs) in cytologic specimens is a morphologic curiosity. The exact mechanism of formation and composition of these peculiar rings is a mystery. Their morphologic resemblance to parasites is well recognized and illustrated. Their association with calcium oxalate-like crystals and their presence in a tubercular lymph node are described for the first time below. CASES: Giemsa-stained aspiration smears from an enlarged lymph node and two breast lumps showed purple rings of variable morphology resembling ova, larvae or adult parasites. Misdiagnosis of parasites was avoided by careful evaluation. In all three cases the LRs were associated with calcium oxalate-like crystals and were nonbirefringent. CONCLUSION: LRs can be mistaken for ova, larvae or adult parasites. Awareness of their varied morphology is helpful in avoiding misinterpretation and overdiagnosis of parasitic lesions. The coexistence of LRs and crystals was an unusual finding in this study. PMID- 10834006 TI - Fine needle aspiration of tophi for crystal identification in problematic cases of gout. A report of two cases. AB - BACKGROUND: The definitive diagnosis of gout is best established by demonstration of monosodium urate (MSU) crystals in the synovial fluid or biopsy. Fine needle aspiration cytology (FNAC) of tophi can play a crucial role in diagnosis. CASES: A 36-year-old chronic alcoholic male developed subcutaneous nodules on both malleoli without a history of arthropathy and with normal serum uric acid levels. FNAC of the nodules demonstrated stacks and sheaves of needle-shaped crystals of MSU. A 50-year-old diabetic male developed multiple nodules on the feet. He gave a past history of painful athropathy. A roentgenogram of the feet was suspicious for gout; however, joint aspiration failed, and the serum uric acid levels were normal. At this juncture FNAC of the feet tophi clinched the diagnosis of gout. In both cases, polarization of needle washings (wet mount) and the fixed, Papanicolaoustained smears showed negatively birefringent, needle-shaped crystals of MSU, thus confirming the diagnosis of gout. CONCLUSION: FNAC of gouty tophi is an easy alternative to synovial biopsy and joint fluid analysis. It is simpler, easier and less painful. As crystals are preserved in stained smears, they can be employed for polarization and confirmation of gout. PMID- 10834007 TI - Intraoperative cytodiagnosis of metastatic brain tumors confused clinically with brain abscess. A report of three cases. AB - BACKGROUND: Cystic lesions of the brain may have diverse etiologies, ranging from true cysts to malignant tumors with cystic degeneration. Preoperative determination of the exact nature of them as well as intraoperative diagnosis may be sometimes difficult or even impossible. Sensitivity and specificity of diagnosis will be improved by introducing new methods or combining traditional procedures. CASES: Three metastatic brain carcinomas with primary sites of breast, pancreas and prostate presented as cystic lesions and were confused clinically with abscess. Intraoperative frozen section was not revealing. Cytologic study of sediments of aspirated fluid uncovered malignant cells. CONCLUSION: When combined with frozen section, intraoperative cytologic studies in the form of crush preparation, fine needle aspiration or evaluation of aspirated fluid in cystic lesions (as in our cases) can improve diagnostic accuracy by detecting important diagnostic features that otherwise may be missed. PMID- 10834008 TI - Catheterized urine cytology of mucinous carcinoma arising in the renal pelvis. A case report. AB - BACKGROUND: Primary mucinous carcinoma of the renal pelvis is a rare tumor; therefore, criteria for cytologic diagnosis of this tumor have not been established. CASE: An 81-year-old woman suffered from macrohematuria for six months and was found to have a tumor in the right kidney by radiographic examination. Catheterized urine obtained from the right renal ureter was viscous and contained spherical clusters of cells with occasionally vacuolated, lacy and basophilic cytoplasm. In the small to medium-sized nuclei, chromatin was coarse and granular, and the nuclear membrane was thin and nearly smooth. Large nucleoli were evident in some of the nuclei. These findings were consistent with adenocarcinoma possibly of mucinous type. CONCLUSION: Preoperative diagnosis of mucinous carcinoma is possible by cytologic findings of catheterized urine together with clinical data. PMID- 10834009 TI - Fine needle aspiration cytology of primary extraskeletal myxoid chondrosarcoma. A case report. AB - BACKGROUND: Extraskeletal myxoid chondrosarcoma is a rare soft tissue tumor of the extremities. Since it usually lacks obvious chondroid differentiation on light microscopy, it needs to be distinguished from other myxoid soft tissue sarcomas. CASE REPORT: The diagnosis of extraskeletal myxoid chondrosarcoma was made on fine needle aspiration in a patient with a swelling in the right calf. Cellular myxoid fragments having round to oval cells with grooved nuclei arranged in a cordlike pattern suggested chondroid differentiation. The diagnosis was confirmed by histopathology. CONCLUSION: Fine needle aspiration cytology can be diagnostic of extraskeletal myxoid chondrosarcoma even in the absence of obvious chondroid differentiation. PMID- 10834010 TI - Fine needle aspiration cytology of mesenchymal hamartoma of the liver. A case report. AB - BACKGROUND: Mesenchymal hamartoma (MH) of the liver constitutes the third or fourth most common tumor of the liver in childhood and occurs most commonly in the first two years of life. MHs of the liver are seldom aspirated, and reports on the role of fine needle aspiration (FNA) in the diagnosis of MH are scarce. Clinically, cytologically and even histologically, MH can be mistaken for a number of reactive and neoplastic hepatic lesions that may occur in children under 2 years of age. CASE: A 10-month-old Pakistani female presented with a history of a right-sided, nonpainful abdominal swelling. Abdominal computed tomography showed a large, partly solid and partly cystic, heterogeneous hepatic mass. FNA cytology showed clusters of both epithelial and mesenchymal/spindle shaped cells with pieces of loose connective tissue. A cytologic differential diagnosis of mesenchymal hepatic hamartoma and hepatoblastoma of the possible mixed mesenchymal/epithelial subtype was rendered. The histopathologic diagnosis of the resected tumor mass was benign mesenchymal hamartoma of the liver. CONCLUSION: In children under 2 years of age who present with partly solid and partly cystic hepatic masses, the possibility of MH of the liver should be considered. FNA has a role in the diagnosis of MH. The cytopathologist should be aware of the patient's age, radiologic features and cytologic appearances of this rare, benign neoplasm. Histologic examination of tru-cut biopsies and immunohistochemical stains can help to exclude other pediatric neoplasms that may show cytologic features similar to or mimicking those of MH. PMID- 10834011 TI - Solitary luteinized follicle cyst of pregnancy. Report of a case with cytologic findings. AB - BACKGROUND: Giant luteinized follicle cyst is a recently recognized cause of ovarian enlargement during pregnancy and the puerperium. Only rare cases of this clinical condition have been reported in the literature, and the cytologic features have not been previously described. CASE: A 34-year-old, pregnant woman presented at 10 weeks' gestation with a large, right ovarian cyst. Clinical management was initially conservative, but four weeks later the patient presented with acute abdominal pain. At laparotomy, the ovarian cyst was aspirated and a cystectomy performed. On cytologic examination, the presence of a luteinized follicle cyst of pregnancy was suggested. Subsequent histologic examination confirmed the diagnosis. CONCLUSION: This case illustrates the clinicopathologic features of a luteinized follicle cyst of pregnancy with special emphasis on the cytologic characteristics. The cytologic features are sufficient to establish the diagnosis in conjunction with the ultrasonographic appearance. PMID- 10834012 TI - Fine needle aspiration cytology of well-differentiated liposarcoma. A report of two cases. AB - BACKGROUND: Well-differentiated liposarcoma is difficult to diagnose on fine needle aspiration cytology (FNAC) smears and may create considerable diagnostic problems. CASES: Males aged 60 and 45 years presented with a swelling in the groin and retroperitoneal region, respectively. FNAC showed large cells with multilobulated nuclei and mature-looking fat tissue. A soft tissue tumor with bizarre cells was diagnosed cytologically in case 1 and liposarcoma in case 2. Histologically, both cases were diagnosed as well-differentiated sclerosing liposarcoma. CONCLUSION: The cytologic diagnosis of well-differentiated liposarcoma should be done with caution, and the sites should be taken into consideration. Deep-seated tumors with large, bizarre, giant cells should have wide excision as they recur more frequently. PMID- 10834013 TI - Localized tenosynovial giant cell tumor of tendon sheath. A case report. AB - BACKGROUND: Localized tenosynovial giant cell tumor of tendon sheath (TGCT-L) is a benign, slowly growing lesion with a peak incidence in the third to fifth decade of life. It is thought to arise from the synovium of tendon sheaths, frequently affecting interphalangeal joints of the hands, feet, ankles and knees. Although the histopathologic appearances are well established, only a few reports describe the cytomorphology of this lesion. CASE: A 37-year-old female presented with a slowly growing, nontender mass located near the left ankle joint. The cytologic features of localized tenosynovial giant cell tumor of tendon sheath (TGCT-L) include abundant mononuclear histiocytic cells occurring singly and in three-dimensional tissue fragments, hemosiderin within histiocytes and a few multinucleated giant cells. Subsequently, the histopathologic examination of the surgical specimen was proven to be TGCT-L. CONCLUSION: Fine needle aspiration cytology can be used as a diagnostic tool for early and accurate detection of TGCT-L since the cytologic features combined with clinical details are sufficiently distinctive. PMID- 10834014 TI - Fine needle aspiration cytodiagnosis of primary squamous cell carcinoma of the gallbladder. Report of two cases. AB - BACKGROUND: Cytologic descriptions of the diagnosis of the rare primary carcinoma of the gallbladder are sparse. The obstacles to the diagnosis are ascribed to vague symptoms and inaccessibility of the gallbladder to biopsy. We describe two cases of primary squamous cell carcinoma of the gallbladder that were diagnosed using fine needle aspiration (FNA) cytology. CASES: Both patients were elderly females with vague symptoms of abdominal pain, jaundice, loss of weight, anorexia and mild fever. Imaging studies showed enlargement of the gallbladder with a soft tissue mass in the fundus and abnormally thickened wall, indicative of a tumor. Also, in case 1, gallstones and enlarged, matted paraaortic and porta hepatis lymph nodes were present. FNA of the gallbladder was done after selection of a suitable puncture site and needle positioning, which was confirmed by computed tomography. The aspirated material was collected as needle and syringe washings, and from half the material filter preparations were made and stained for cytologic study, while the remainder was used for making cell blocks for histologic study. Both cases showed cytohistologic features of a moderately well differentiated, necrotizing squamous cell carcinoma characterized by keratinizing malignant cells with orangeophilic cytoplasm. In both cases immunostaining for high-molecular-weight keratins (AE1/AE3) and cytokeratin (CAM 5.2) was positive, while staining for carcinoembryonic antigen, B72.3 and other markers was negative. CONCLUSION: This study further confirmed that with the widespread use of more-sophisticated imaging techniques, the gallbladder is becoming more readily accessible to visualization. In view of this, FNA cytodiagnosis holds promise as a noninvasive technique in the diagnosis of gallbladder neoplasms. PMID- 10834015 TI - Inflammatory pseudotumor of the kidney: report of a case with fine needle aspiration cytology. PMID- 10834016 TI - Microglandular hyperplasia of the uterine cervix: cytologic diagnosis in cervical smears. PMID- 10834017 TI - Pinus pollen in a vaginal smear. PMID- 10834018 TI - CA-15.3 immunostaining in mesothelial disorders. PMID- 10834019 TI - Trichomonas vaginalis in urine cytology. PMID- 10834020 TI - Differential diagnosis of mesothelial and ovarian cancer cells in ascites by immunocytochemistry using Ber-EP4 and calretinin. PMID- 10834021 TI - Fine needle aspiration cytology of adenomyoepithelioma of the breast: a case indistinguishable from phyllodes tumor in cytologic findings and clinical behavior. PMID- 10834022 TI - Clinical significance of cell "cannibalism". PMID- 10834023 TI - Malignant oncocytoma of a submandibular salivary gland. PMID- 10834024 TI - Effects of calcium and vitamin supplementation on colon cell proliferation in colorectal cancer. AB - Calcium and antioxidant vitamins, such as A, C, and E, have been shown to reduce colorectal epithelial proliferation and thereby to act as possible chemoprotective agents in colorectal cancer. We investigated the effects of an intervention with calcium and vitamins on cell proliferation in the colonic mucosa of patients operated on for colorectal cancer. Patients with resected colorectal cancer Dukes' stage B-C were randomized to receive daily 30,000 IU of axerophthol palmitate (vitamin A) plus 1 g ascorbic acid (vitamin C) plus 70 mg of dl-alpha-tocopherol acetate (vitamin E) and 2 g natural calcium daily or indistinguishable placebo for 6 months. At the time of surgery and after 6 and 12 months of treatment, cell kinetics of normal colonic mucosa were assessed by using proliferating cell nuclear antigen (PCNA). Ninety patients were enrolled and 77 were assessable: 34 in the treatment group and 43 in the placebo group. A significant reduction of mean total PCNA labeling index (PCNALI) was evident in both groups after 6 months (vitamins/calcium, from 16.11 +/- 2.43 to 10.71 +/- 2.81; placebo, from 17.30 +/- 2.63 to 12.53 +/- 3.40). The difference in the percentage of reduction of mean PCNALI between baseline and after 6 months was not statistically significant in the treatment and placebo groups: 34% and 28%, respectively. A second control, 6 months after discontinuation of vitamin and calcium supplementation, showed a further decrease of mean total PCNALI in both groups, but this was not statistically significant. Our randomized trial showed that calcium and vitamin supplementation does not reduce cell kinetics of colon epithelium. Furthermore, this study suggests the need for extreme caution in the interpretation and publication of studies on chemoprotectants in colon cancer without a control group. PMID- 10834025 TI - Interferon alpha-2a and 13-cis-retinoic acid in patients with metastatic renal cell cancer. AB - Treatment of patients with metastatic renal cell cancer (RCC) with interferon alpha-2a (IFN) and 13-cis-retinoic acid (CRA) was first reported to be tolerable on an outpatient basis and to yield a 30% objective response rate. We sought to confirm these preliminary results by conducting a phase II trial of therapy with IFN/CRA in patients with bidimensionally measurable RCC. Twenty-five patients were enrolled. The median age was 58 (range, 47-75 years) and the median Karnofsky performance status was 90 (range 60-100). Seventeen patients (60%) had undergone prior nephrectomy and none had received prior systemic therapy. Treatment consisted of oral CRA at 1 mg/kg/day and IFN self-administered by subcutaneous injection at 3 MU/day with weekly escalation to 6 and 9 MU/day. Treatment was well tolerated, with cheilitis, influenza-like symptoms, and fatigue the most common toxicities. Severe toxicity was reversible and consisted of grade 4 cheilitis in one patient and grade 3 malaise/fatigue in two patients. One complete and four partial responses were observed, for an objective response rate of 20% (95% confidence interval, 4-36%). We conclude that treatment with CRA/IFN for RCC is tolerable on an outpatient basis and induces objective responses in some patients. The contribution, if any, of CRA to the responses observed will be determined in ongoing randomized phase III trials. PMID- 10834026 TI - Phase I study of weekly vinorelbine in combination with weekly paclitaxel in adult patients with advanced refractory cancer. AB - Vinorelbine and paclitaxel are highly active antineoplastic agents. Preclinical data indicate a potential for antitumor synergy for a number of common tumor types when they are combined. We investigated a novel weekly schedule of both agents. Eighteen patients with advanced cancer were entered onto this phase I trial. Vinorelbine and paclitaxel were given weekly in combination for 6 consecutive weeks, followed by a 2-week break. Sequential cohorts of patients were treated at two dose levels: vinorelbine 22.5 mg/m2 followed by paclitaxel 40 mg/m2 and vinorelbine 22.5 mg/m2 followed by paclitaxel 60 mg/m2. Ten patients completed at least one 8-week course of therapy. Neutropenic myelosuppression was dose limiting at level II. Neurotoxicity was not dose limiting. Objective responses were seen in patients with esophageal, lung, and breast cancer and suggest that this is an active regimen worthy of further investigation in selected diseases. Phase II trials of this regimen are in progress. PMID- 10834027 TI - Comparative study of sequential combinations of paclitaxel and methotrexate on a human bladder cancer cell line. AB - We investigated the cytotoxic effect of different sequential combinations of paclitaxel (Taxol, TX), an antimicrotuble agent, and methotrexate (MTX), a potent inhibitor of dihydrofolate reductase, on a human bladder cancer cell line (T24). The results obtained with a colony-forming assay 5 days after 24 hr of exposure to TX alone showed a strong decline in colony growth up to a concentration of 6 nM. There was a plateau phase at TX concentrations ranging from 6 to 10 nM. The IC50 values found with the different dosing sequences and 24-hr exposure to the drugs were 0.24 microM for MTX alone, 3.49 nM for TX alone, 3.84 nM for TX followed by MTX, 3.75 nM for TX and MTX simultaneously, and 2.10 nM for MTX followed by TX. The cytotoxic effect due to drug interactions in the different sequential combinations was evaluated by an algebraic method. All the combination sequences were found to be synergistic (combination index [CI] < 1) at low TX concentrations. An antagonistic effect (CI > 1) was obtained at higher TX concentrations. Our data demonstrate that the T24 cell line is sensitive to TX and that the cytotoxic effect is enhanced to different degrees depending on the treatment sequence when TX is combined with MTX. These findings could have implications in the design of appropriate administration schedules of these two drugs for the treatment of bladder cancer. PMID- 10834028 TI - Cancer clinical trials and subject use of natural herbal products. PMID- 10834029 TI - High-dose chemotherapy and autologous stem cell transplantation for breast cancer. PMID- 10834030 TI - Historical overview of biologic response modifiers. PMID- 10834031 TI - Use of hair dyes, hematopoietic neoplasms, and lymphomas: a literature review. II. Lymphomas and multiple myeloma. AB - We review studies on hair dyes and lymphomas and multiple myeloma (MM). A computerized literature search for the years 1966 through 1996 was conducted. Data were extracted using a standardized form that recorded study design, study population, type of cases, comparison group, sources of data on personal exposure to hair dyes, method of data collection, type of exposure data collected, covariates, and results. This review identified 10 epidemiologic studies published in the English literature that examined personal use of hair dyes and lymphomas or MM. These studies include three evaluations of Hodgkin's disease, five of non-Hodgkin's lymphoma (NHL), two of lymphomas with type not specified, and six of MM. For Hodgkin's disease, one case-control study reported some positive associations with use of permanent hair dyes, whereas two cohort studies found no associations with ever use of hair dyes. For NHL and MM, several evaluations suggest associations with use of permanent dyes, particularly with duration, frequency, age at first use, and dark colors. However, these associations are not consistent within and between studies. For lymphomas with type not specified, one study was superseded by a more recent report with NHL specific data and a second study was limited by small numbers of exposed subjects. At this time, it is not possible to determine if the inconsistent associations between permanent hair dyes and NHL and MM reflect sampling variability or differences in methods between studies. Because an appreciable fraction of the population has potential exposure to permanent hair dyes, elucidation of such issues may be warranted with studies that include adequate numbers of exposed subjects and that elicit information on personal use of hair dyes over time. PMID- 10834032 TI - Natural and induced human antibody response to cancer. PMID- 10834033 TI - Renal cancer immunotherapy: a ray of hope or regression to the mean? PMID- 10834034 TI - High-dose chemotherapy with autologous stem cell rescue for malignant brain tumors. PMID- 10834035 TI - High-dose chemotherapy for breast cancer: we have not heard the final word. PMID- 10834036 TI - Taxane-based therapy for breast cancer: combination or sequential therapy? PMID- 10834037 TI - Scientific reporting of ethnicity, age, sex and race. PMID- 10834038 TI - Interpreting the results of small trials. PMID- 10834039 TI - Does the urea breath test tell us what we need to know? PMID- 10834040 TI - Distorted spending priorities in Canada. PMID- 10834041 TI - TB among aboriginal Canadians. PMID- 10834042 TI - Treating TB in the 1950s. PMID- 10834043 TI - Illness outbreak associated with Escherichia coli O157:H7 in Genoa salami. E. coli O157:H7 Working Group. AB - BACKGROUND: An outbreak of Escherichia coli O157:H7 infection was identified in the spring of 1998, with a 7-fold increase in the number of laboratory-confirmed E. coli O157:H7 cases in southern Ontario. This prompted an intensive investigation by local, provincial and federal public health officials. METHODS: Case interviews of 25 people from southern Ontario were conducted using a broad food history and environmental exposure survey. Laboratory investigations involved both case and food sampling. Specimens of foods sold locally and reportedly consumed by those affected were tested. Common suppliers of suspected foods were identified by cross-referencing suppliers' lists with stores frequented by those who fell ill. A case-control study involving 25 cases and 49 age-matched controls was conducted. This was followed by a comprehensive environmental investigation of the meat processing plant identified as the source of the E. coli. RESULTS: Thirty-nine outbreak-related cases occurred between April 3 and June 2, 1998. Of the 36 case specimens tested all were positive for E. coli O157:H7. The case-control study identified Genoa salami as the most probable (odds ratio 8 [confidence interval 2-35]) source of the outbreak. Samples of Genoa salami produced by the most commonly identified supplier later tested positive for E. coli O157:H7, and the pathogen matched the same pulsed field gel electrophoresis pattern and phage type of the case specimens. INTERPRETATION: Our investigation, which led to a national recall of the brand of dry fermented Genoa salami identified as the source of the outbreak, supports an adherence to stringent manufacturing requirements for fermented meat products. A review of the Canadian standards for fermented meat processing and the effectiveness of their implementation is warranted. PMID- 10834044 TI - Treatment and outcomes of community-acquired pneumonia at Canadian hospitals. AB - BACKGROUND: Community-acquired pneumonia is a common disease with a large economic burden. We assessed clinical practices and outcomes among patients with community-acquired pneumonia admitted to Canadian hospitals. METHODS: A total of 20 hospitals (11 teaching and 9 community) participated. Data from the charts of adults admitted during November 1996, January 1997 and March 1997 were reviewed to determine length of stay (LOS), admission to an intensive care unit and 30-day in-hospital mortality. Multivariate analyses examined sources of variability in LOS. The type and duration of antibiotic therapy and the proportion of patients who were treated according to clinical practice guidelines were determined. RESULTS: A total of 858 eligible patients were identified; their mean age was 69.4 (standard deviation 17.7) years. The overall median LOS was 7.0 days (interquartile range [IQR] 4.0-11.0 days); the median LOS ranged from 5.0 to 9.0 days across hospitals (IQR 6.0-7.8 days). Only 22% of the variability in LOS could be explained by known factors (disease severity 12%; presence of chronic obstructive lung disease or bacterial cause for the pneumonia 2%; hospital site 7%). The overall 30-day mortality was 14.1% (95% confidence interval [CI] 11.8% 16.6%); 13.6% of the patients were admitted to an intensive care unit (95% CI 11.4%-16.1%). The median duration of intravenous antibiotic therapy was 5 days (range 3.0-6.5 days across hospitals). Although 79.8% of patients received treatment according to clinical practice guidelines, the rate of compliance with the guidelines ranged from 47.9% to 100% across hospitals. INTERPRETATION: Considerable heterogeneity exists in the management of community-acquired pneumonia at Canadian hospitals, the causes of which are poorly understood. PMID- 10834045 TI - Postmenopausal estrogen replacement therapy and increased rates of cholecystectomy and appendectomy. AB - BACKGROUND: Several studies have indicated that estrogen may prime inflammatory and nociceptive pathways, leading to symptoms that mimic cholecystitis. We set out to confirm the relation between recent estrogen use and cholecystectomy in postmenopausal women and to test the novel hypothesis that a similar relation exists for appendectomy. METHODS: We developed a retrospective cohort using prescribing and surgical procedure information from health administrative databases for approximately 800,000 female residents of Ontario who were over 65 years of age between July 1, 1993, and Mar. 31, 1998. We compared the incidence of cholecystectomy and appendectomy among women recently prescribed estrogen replacement therapy, levothyroxine and dihydropyridine calcium-channel antagonists (DCCA) using age-adjusted Cox proportional hazards models. Patients were followed for a mean of 540 (standard deviation [SD] 449) days. RESULTS: Compared with women taking DCCA, those who had recently begun taking estrogen were significantly more likely to undergo cholecystectomy (age-adjusted risk ratio [aRR] 1.9, 95% confidence interval [CI] 1.6-2.2) and appendectomy (aRR 1.8, 95% CI 1.1-3.0). No significant difference in either outcome measure was found between the levothyroxine users and the DCCA users. INTERPRETATION: This study identifies an increased risk of cholecystectomy and appendectomy among postmenopausal women who have recently begun estrogen replacement therapy. PMID- 10834046 TI - The frustrations of fighting foodborne disease. PMID- 10834047 TI - Screening for lung cancer: can it be cost-effective? AB - Last year, the Lancet reported on a study concerning a particular, avant-garde regimen of CT-based screening for lung cancer, showing its great superiority relative to the corresponding regimen based on traditional radiography (Lancet 1999;354:99-105). That report was met with great interest in the media, worldwide. It thereby also led to substantial public demand for the state-of-the art screening, in the United States especially. I here argue that, despite the prevailing official recommendations against any and all screening for lung cancer in the United States and Canada, it actually already is knowable that modern screening, with suitable specifications of both the screening itself and its recipient, not only is effective but can be effective enough to amply justify its cost. It thus is time for authorities to formulate, ever more inclusively, those cost-justifying specifications--and to promote research providing for further expansions of and innovations in them. American authorities, however, have not reacted this way to the new situation and, I dare say, they have been tenaciously irrational (and thereby irresponsible) in their underlying ideas about the nature of the proper knowledge-base for screening practice and of the research serving to advance this. It remains to be seen how timely, and how compellingly rational, the Canadian official reactions will be; and this too matters greatly, as countless lives hang in the balance, within Canada and, especially, in the world at large. Ex Canada lux? PMID- 10834048 TI - Recommendations for the management and treatment of dyslipidemia. Report of the Working Group on Hypercholesterolemia and Other Dyslipidemias. PMID- 10834049 TI - Carotid angioplasty and stenting: current status. AB - Carotid angioplasty and stenting has recently emerged as a popular alternative to endarterectomy for the treatment of carotid atherosclerosis. Carotid endarterectomy has been scientifically validated, but many believe carotid angioplasty and stenting to be a less invasive, less expensive and equally safe and effective method of treatment. The evidence for and against the use of each procedure will be discussed. PMID- 10834050 TI - Blood collecting resumes in Newfoundland community. PMID- 10834051 TI - Nova Scotia launches assault on assault. PMID- 10834052 TI - Physicians fight for access to tobacco info, hope to show criminal negligence. PMID- 10834053 TI - Indoor moulds and human health. PMID- 10834054 TI - OMA fights rising tuition fees with bursaries, other provinces to follow suit. PMID- 10834055 TI - Canada hopes to climb the DNA ladder to success. PMID- 10834056 TI - Holistic approach can get to all of the root causes. PMID- 10834057 TI - Who's minding the hospital? Absenteeism soars. PMID- 10834058 TI - Radiology on the rebound. PMID- 10834059 TI - Is an e-commerce gap emerging among the nation's hospitals? PMID- 10834060 TI - To the rescue. PMID- 10834061 TI - Building excellence. 1999 Vista Awards. PMID- 10834062 TI - Nova Award winners 2000. PMID- 10834063 TI - Inhibition of metabolism and growth of Mycobacterium leprae by gamma irradiation. AB - Mycobacterium leprae is uncultivable on artificial medium, but viability can be maintained without multiplication for a limited time in vitro. In this study, we evaluated gamma-irradiation (gamma-irr) as a means to kill this slowly growing organism. Freshly harvested, viable, athymic, nu/nu mouse-derived M. leprae were exposed to varying doses of gamma-irr from a 60Co source. Two indicators of bacterial viability were determined: metabolism, measured by oxidation of 14C palmitic acid to 14CO2 in the BACTEC 460 system, and multiplication, measured by titration in the mouse foot pad. gamma-Irr of both M. leprae and M. lufu, a cultivable control mycobacterium, resulted in a dose-dependent inhibition of viability. gamma-Irr of up to 10(3) rad had little effect on the metabolic activity of either organism. For M. leprae, 10(4)-10(5) rad caused an intermediate inhibitory effect; whereas 10(6) rad yielded almost total inhibition. In the mouse foot pad assay, up to 10(4) rad had little effect on M. leprae growth; however, 10(5) rad resulted in at least a 2-log reduction in the number of bacilli recovered and no M. leprae growth was measurable after exposure to 10(6) rad. With M. lufu, 10(5) rad inhibited metabolic activity by 99% and caused > or = 2-log reduction in the number of colony forming units (CFU). No CFU of M. lufu were recovered after exposure to 10(6) rad. Scanning electron microscopy revealed the presence of some aberrant protrusions on the cell surface of lethally irradiated M. leprae; whereas boiling and autoclaving caused obvious morphological denaturation. These data suggest that gamma-irr is an effective way to kill M. leprae without causing extensive damage to the cell architecture. Killing M. leprae by gamma-irr may be preferable when comparing cellular responses to live versus dead bacilli in vitro and in vivo. PMID- 10834064 TI - The effect of ultraviolet light radiation on Mycobacterium leprae. AB - Ultraviolet (UV) light is recognized as a potent sterilizing aid, but its relative effectiveness against Mycobacterium leprae has not been shown. We examined the influence of UV on the growth and metabolic activity of M. leprae harvested fresh from foot pads of nude mice. Temporary static suspensions were exposed to timed intervals of UV radiation generated from a fixed source to constitute dosages ranging from 0-12.64 x 10(4) erg/cm2. The metabolic activity of the bacilli was indexed by the oxidation of 14C-palmitate in BACTEC 12-B vials. The long-term effects of irradiation on cell division and growth were assessed by inoculation of BALB/c mouse foot pads. The metabolic activity in BACTEC showed an immediate dose-response-related decline to a maximum of 50% of the control activity after exposure to 6.3 x 10(4) erg/cm2. Mouse foot pad studies showed a similar dose-response pattern. Effective-dose determinations based on metabolic or foot pad data were similar. UV doses of 3.52 x 10(4) erg/cm2 resulted in an average 50% killing, and 7.73 x 10(4) erg/cm2 killed 84% of the M. leprae exposed. This UV sensitivity is similar to that reported for M. tuberculosis. UV sterilization and disinfection practices suitable for M. tuberculosis are likely to be equally effective for M. leprae. PMID- 10834066 TI - Pathology of a lepromatous eye. AB - Histopathological examination of an enucleated eye from a lepromatous leprosy patient showed the cornea, ciliary body, and part of the choroid to be infiltrated by macrophages filled with Mycobacterium leprae. The walls of blood vessels in the sclera, ciliary body and the anterior choroid demonstrated the presence of M. leprae, giving credence to the blood-borne entry of M. leprae into the eye. Unlike the eyes of experimental animals infected with M. leprae, histopathological study of this eye from a lepromatous leprosy patient demonstrated that M. leprae, although demonstrable in the anterior choroid, could not be found in the posterior parts of the eye, substantiating the claim that leprosy does not affect the posterior parts of the eye directly. PMID- 10834065 TI - Comparative study of Mitsuda reaction to nude mouse and armadillo lepromin preparations using nine-banded armadillos. AB - In 14 nine-banded armadillos the Mitsuda response to nude mouse-derived lepromin (lepromin-nu/nu) was compared to that of armadillo-derived lepromin (lepromin-A) by injecting the reagents intradermally into either side of the abdomen of the animal and examining the biopsies from the sites after 12 days. The histopathologic responses to both antigens were found to be similar, whether the animal was Mitsuda-negative (lepromatous) or Mitsuda-positive (tuberculoid). It is pointed out that armadillos are good experimental models for leprosy, and their use can replace humans in experimental studies. PMID- 10834067 TI - Antileprosy protective vaccination of rhesus monkeys with BCG or BCG plus heat killed Mycobacterium leprae: immunologic observations. AB - Groups of rhesus monkeys were vaccinated and boosted with Mycobacterium bovis bacillus Calmette Guerin (BCG) or BCG plus low-dose (LD) or high-dose (HD) heat killed M. leprae (HKML), or were unvaccinated. Prior to and following vaccination boosting and subsequent M. leprae (ML) challenge, these and unvaccinated, unchallenged control monkeys were observed longitudinally for approximately 3 years. Vaccination with BCG plus HKML initially stimulated significant in vitro blood mononuclear cell blastogenic responses to lepromin, which returned to baseline post-boosting and post-live-ML-challenge, minimally reappearing significantly 2 years post-ML-challenge. Vaccination with BCG failed to stimulated positive blastogenic responses to lepromin before ML-challenge but small, marginally positive, intermittent responses were seen post-ML-challenge. Compared to the unvaccinated ML-challenged group, significant increases in the numbers of blood CD4+ and CD8+ T-cell subsets and an increased CD4+:CD8+ ratio were observed in both BCG plus HKML-vaccinated, ML-challenged groups, but not in the BCG-only-vaccinated, ML-challenged group. CD4+CD29+ and CD4+CD45RA+ subset numbers increased significantly over time in only the BCG plus LD HKML vaccinated, ML-challenged group. Compared to unvaccinated, ML-challenged groups, vaccination with BCG or BCG plus HKML followed by ML-challenge produced lower IgM:IgG antiphenolic glycolipid-I (PGL-I) serum antibody ratios and protected rhesus monkeys from clinical leprosy, consistent with prior observations that low IgM:IgG anti-PGL-I responses correlated with resistance to and protection from leprosy. PMID- 10834068 TI - Interferon-gamma responses to candidate leprosy skin-test reagents detect exposure to leprosy in an endemic population. AB - New tools for the detection of leprosy exposure in a community will be necessary for the eradication of leprosy. Candidate leprosy skin-test antigens derived from the fractionation of the leprosy bacillus into cytoplasmic and cell-wall proteins free of immuno-inhibitory mycobacterial lipoglycans and carbohydrates were used in an overnight blood test to determine whether exposure to leprosy can be detected by the production of the cytokine interferon gamma (IFN-gamma). Strong IFN-gamma responses were detected in leprosy contacts to both skin-test antigens compared with control subjects from the same endemic communities. There was little response in patients with tuberculosis. Responses were greatest in contacts with recent leprosy exposure. The implications of these findings for the application of these reagents in a field trial as skin tests to detect exposure to leprosy are discussed in light of the strong association between overnight IFN gamma to PPD and the tuberculin skin-test responses previously reported. PMID- 10834069 TI - Human leukocyte antigens in forms of leprosy among Japanese patients. AB - Human leukocyte antigens (HLA) class II alleles were analyzed among Japanese leprosy patients to ascertain whether immunogenetic differences exist among the leprosy classification forms of Ridley and Jopling. Ninety-three unrelated Japanese leprosy patients (21 lepromatous, 24 borderline lepromatous, 17 mid borderline, 26 borderline tuberculoid, 5 tuberculoid) and 114 healthy control subjects were investigated. The frequencies of HLA-DRB1*1501, -DRB5*0101, DQA1*0102 and DQB1*0602 were significantly increased in all of the Japanese leprosy patients. The frequencies of HLA-DRB1*0405, -DQA1*03 and -DQB1*0401 were significantly decreased in the Japanese patients after correction of the p value. Conversely, there were no significantly different distributions of the HLA-DRB1, DRB5, -DQA1, DQB1 alleles in the five subgroups of these patients. We conclude that HLA class II alleles were not associated with the form of leprosy. Other HLA, a non-HLA gene, and/or environmental factors may play a critical role in the different manifestations of leprosy. PMID- 10834070 TI - Epidemiology of leprosy in Taiwan; its pattern in children. AB - OBJECTIVE: To study the epidemiology of leprosy in children in Taiwan. SETTING: Taiwan, with a population increase from 3.3 to 21.7 million, several tides of immigration and national leprosy control programs, from 1910 to 1997. DESIGN: To collect and analyze the documents of Taiwan leprosy surveys and charts of the National Leprosy Control Center. PATIENTS AND MEASUREMENTS: Cumulative and new number of all-age and pediatric-age patients, prevalence rates, new case detection rates, and results of skin bacterial smears. RESULTS AND CONCLUSIONS: The prevalence rates of all-age leprosy ranged between 1.54 and 3.22 per 10,000 population. The proportion of children among all-age patients reached the highest of 4.93% in 1966, dropping to 0% in 1984 and thereafter, until 1988 and 1991 when two and one pediatric-age patients appeared, respectively, following the influx of immigrants from leprosy-endemic countries. The rise and fall of new patients younger than 15 years and 15 years or older were slightly correlated (r = 0.935, p < 0.001). Detection and confirmation of leprosy in children are usually belated. Physicians should still be acquainted with the clinical diagnosis of leprosy since sporadic cases of leprosy can reappear, particularly among children coming from endemic countries. PMID- 10834071 TI - Clarification to Rada-Schlaefli, et al. PMID- 10834072 TI - Assessment of skin temperature by palpation in leprosy patients: interobserver reliability and correlation with infrared thermometry. PMID- 10834073 TI - HIV seroprevalence in leprosy patients. PMID- 10834074 TI - The effect of oil of hydnocarpus on excision wounds. PMID- 10834075 TI - Phagocytic myeloperoxidase in leprosy pathogenesis. PMID- 10834076 TI - Effects of lead on birds (Laridae): a review of laboratory and field studies. AB - Lead is one of the most common metals in contaminated ecosystems. Although lead poisoning and mortality have long been known, little is known of the behavioral effects produced by low levels of lead in wild animals. Herein a 15-yr research program on the behavioral effects of lead using herring gulls (Larus argentatus) and common terns (Sterna hirundo), referred to as larids, as models is reviewed. The doses used in laboratory studies were sufficient to produce lead concentrations in feathers that were equivalent to those found in some birds living in the wild. The exposure consisted of a single or multiple intraperitoneal (i.p.) injection of lead acetate. Both dose and day of exposure influenced behavioral development in young larids, with most effects increasing with dose, and decreasing with age. Low-level lead affected growth, locomotion, balance, food begging, feeding, thermoregulation, depth perception, and individual recognition in laboratory and in wild birds. The accuracy of individual recognition was most affected by lead exposure from 2 to 6 d of age; exposure at 12 d did not affect accuracy, but it delayed response time significantly. Behavioral deficits observed in lead-injected young in the wild were similar to those observed in the laboratory, except that recovery was more complete by fledging than it was in laboratory-raised chicks. Further, parents in the wild were able to provision lead-exposed chicks sufficiently so that they fledged at similar weights as control chicks, a feat that is difficult to achieve in the laboratory. The lead-induced behavioral deficits observed in the laboratory and in the wild are sufficient to affect growth and survival in wild herring gulls. Lead treatment altered the expression of cell adhesion molecules (CAMs), which play a crucial role in the formation and deployment of neurons in the developing brain. The timing and sequencing of CAM expression is critical to normal development, and the different consequences of lead exposure at different ages may be related to interference at different points in the sequence. PMID- 10834077 TI - Use of response biomarkers in milk for assessing exposure to environmental contaminants: the case for dioxin-like compounds. AB - Screening for environmental contaminants in milk is generally conducted by chemical analysis, yet such an approach may be time-consuming and expensive, and is not indicative of the physiological consequences of such exposure. The focus of this review is to summarize those constituents of milk that may be altered by maternal exposure to one of the most biologically active environmental pollutants, 2,3,7,8-tetrachlorodibenzo-p-dioxin, and to develop the case for their use as biomarkers of response. Several chemical and/or cellular components of milk are potentially useful as biomarkers, and may be developed as convenient, biologically relevant indicators of maternal exposure to dioxin-like compounds. PMID- 10834079 TI - Employer-sponsored health insurance: are employers good agents for their employees? AB - Employers in the United States provide many welfare-type benefits, such as life insurance, disability insurance, health insurance, and pensions, to their employees. Employers can be viewed as performing an agency role in purchasing pension, health, and other welfare benefits for their employees. An exploration of their competence in this role as agents for their employees indicates that large employers are very helpful to their employees in this arena. They seem to contribute to individual employees' welfare by providing them with valued services in purchasing health insurance. PMID- 10834078 TI - Immunomodulation by fungal toxins. AB - The availability of immunotoxicity data for fungal toxins varies considerably for different toxins. The following is a comprehensive review of the most recent literature on the immunotoxicity of aflatoxins, fumonisins, gliotoxin, ochratoxins, patulin, and trichothecenes. Aflatoxin is an immunomodulating agent that acts primarily on cell-mediated immunity and phagocytic cell function. In addition to further characterization of aflatoxin-induced immunotoxicity in various species, some recent studies have focused on ameliorating the effects of aflatoxin by supplementing or amending the diet. The immunomodulatory effects of ochratoxins have also been considered for many years. Notably, recent studies have examined immune function in the offspring of rats and mice exposed to ochratoxin pre- and perinatally. Fumonisin toxicity has been characterized relatively recently in comparison to aflatoxin and ochratoxin, and fumonisin induced immunotoxicity is an area of active research. As these studies progress, they may also clarify the role of sphingolipid metabolism in immune function. The most recent study of patulin immunotoxicity in mice indicates that exposure to levels found in foods and feeds would not likely result in immunotoxicity. Exposure to gliotoxin would most likely be by infection with gliotoxin-producing fungi. Although the toxin is immunosuppressive in vitro, the link between immunosuppression and the presence of gliotoxin in infected tissues in vivo has yet to be made. The trichothecenes can both suppress and stimulate immune function. By comparison, more information is available on the molecular events associated with trichothecene-induced immunomodulation than for any other fungal toxins. The molecular basis of immune function modulation by fungal toxins remains a frontier for future research. PMID- 10834080 TI - The role of independent agents in the success of health insurance market reforms. AB - The impact of reforms on the health insurance markets cannot be understood without more information about the role played by insurance agents and a closer analysis of their contribution. An in-depth, qualitative study of insurance market reforms in seven illustrative states forms the basis for this report on how agents help to shape the efficiency and fairness of insurance markets. Different types of agents relate to insurers in their own ways and are compensated differently. This study shows agents to be almost uniformly enthusiastic about guaranteed-issue requirements and other components of market reforms. Although insurers devise strategies for manipulating agents in order to avoid undesirable business, these opportunities are limited and do not appear to be seriously undermining the effectiveness of market reforms. Despite the layer of cost that agents add to the system, they play an important role in making market reforms work, and they fill essential information and service functions for which many purchasers have no ready substitute. PMID- 10834081 TI - Do consumers use information to choose a health-care provider system? AB - This study examines the use of information by employees in the Buyers Health Care Action Group, a purchasing coalition of large employers in Minneapolis. BHCAG employers contract directly with multiple health-care provider systems and attempt to inform employees about those choices. Shortly after the close of the 1998 open-enrollment period, a survey of 927 BHCAG employees with single-coverage health insurance was conducted. Seventy-six percent of the employees relied on information from their employer when selecting their current care system. Use of information from the employer was positively related to education and years of residence in the Twin Cities. Previous experience with doctors and hospitals in the care system also was a common information source. Older and low-income workers were more likely to use information from advertisements. The survey results suggest that employers can predict which information sources their employees will use. PMID- 10834082 TI - Disability management, employee health and fringe benefits, and long-term disability claims for mental disorders: an empirical exploration. AB - Mental disorders account for a large share of claims and benefit costs in both private and public long-term-disability (LTD) insurance programs. This is the first empirical study to explore factors that may explain variations in private sector LTD claims incidence and cost across groups of employees. Employee fringe benefit arrangements, including patterns of coverage for mental health treatment, are found to be important predictors of incidence rates. Award rates for public disability insurance coverage (SSDI) are also strongly related to claims incidence, suggesting that private LTD is an important pathway to SSDI benefits. Some employee disability-management strategies, such as front-line manager involvement and provision of alternative jobs for employees returning from disability leave, are predictive of lower claims rates and/or costs. PMID- 10834083 TI - The prevalence of hospital health promotion and disease prevention services: good news, bad news, and policy implications. AB - In recent years, American health care has shifted toward an emphasis on population health in communities. National data from the American Hospital Association Annual Survey of Hospitals are used to describe the prevalence of 26 services provided by general hospitals that could contribute to health promotion and disease prevention (HPDP). Cross-sectional descriptive analyses, based on national data sources, linked HPDP services to hospital characteristics, and factor analysis identified significant categories of HPDP activities. The results showed that many specific HPDP services are offered by thousands of hospitals, but prevalence, distribution, and availability of the services are uneven across the size and ownership of hospitals and their communities. Policy initiatives could increase the prevalence of hospitals' health promotion and disease prevention services, thereby improving the health status of their communities. PMID- 10834084 TI - A technique for extraction and thin layer chromatography visualization of fecal bile acids applied to neotropical felid scats. AB - Fecal bile acid patterns have been used successfully to identify scats. Neotropical felid scats are capable of this biochemical identification because they present low concentrations of plant pigments that would interfere in fecal bile acids detection. However, neotropical felid scats have poor quantities of bile acids, so we developed in this work a proper technique for their extraction, visualization and determination. Twenty eighth feces of seven different felid species, collected from Zoological and Wildlife Parks, were dried and pulverized. The procedure for analyzing feces is: Take one g of pulverized feces and shake for 3 hr at room temperature in 20 ml benzene:methanol; filter and evaporate to 5 ml. Spot on TLC plate and develop in toluene:acetic acid:water. Dry and visualize with anisaldehyde. Field collected scats could be identified by the bile acids pattern revealed by this specific technique and, then, used as a source of information for distribution, density and food habits studies. PMID- 10834085 TI - [The physiological characteristics and pharmacology of individual time perception]. AB - The ability of humans and animals to perceiving time intervals is analyzed. The fundamental and applied aspects of the problem are considered, in particular, from the standpoint of solving chronopharmacological tasks. PMID- 10834086 TI - [The nootropic and anxiolytic properties of different doses of piracetam]. AB - The effect of piracetam at various doses on the behavioral and electrophysiological characteristics was studied, including the development of passive and active avoidance conditional reflexes in rats, their behavior in conflict situations, and the transcallosal evoked response (TER) in rabbit brain. In the dose range from 50 to 300 mg/kg, piracetam improved the avoidance performance of both types and produced a dose-dependent increase in the TER amplitude, but did not affect the behavior of rats in conflict situations. As the drug dose was increased to 400-1000 mg/kg, the positive learning influence disappeared (sometimes the effect was even negative) and the TER increase changed to decrease. In contrast, the conflict situation tests revealed pronounced anxiolytic activity of piracetam at elevated doses. Thus, the nootropic and anxiolytic effects of piracetam (and, probably, of the other tranquilizers as well) do not coexist and are significantly shifted relative to one another on the dose scale, being probably realized via different mechanisms. PMID- 10834087 TI - [Lithium oxybutyrate and the rhythmic structure of the actively exploratory behavior and body temperature in rats under constant illumination conditions]. AB - Under constant illumination (LL 24) conditions, lithium hydroxybutyrate (LHB, daily dose 10 mg/kg, 10 days) facilitated the formation of synchronized 24-h rhythms in the active-search (open-field) behavior and body temperature variation in rats, independently of the objective drug administration time. In the dark (DD 24), LHB increased the free-running rhythms and facilitated synchronization of the internal harmonics, especially upon the drug administration in the objective p.m. phase. PMID- 10834088 TI - [A comparative study of the central H-cholinergic-blocking and NMDA-blocking actions of MK-801, memantin, amantadine, pyrilen and IEM-1754 in experiments on intact rats]. AB - Pyrilene acts as a central H-cholinoblocker upon intramuscular injection at a dose of 0.02-0.08 mg/kg, and as an NMDA-blocker, when the dose is increased to 0.2-0.8 mg/kg. Similarly, amantadine exhibits the properties of H-cholinoblocker and NMDA-blocker in the dose intervals 10-15 mg/kg and 120-180 mg/kg, respectively. The activity of MK-801 markedly exceeds that of memantine, although close doses of both NMDA-blockers inhibit the NMDA and corazole effects, as well as the central effects (analgesia and seizure) of nicotine, thus showing no significant selectivity with respect to NMDA. IEM-1754 tested on intact animals exhibited a selective action upon the NMDA receptors, preventing the NMDA-induced analgesia and lethality and the corazole-induced convulsions at doses 10-100 times lower as compared to those preventing nicotine-induced seizure and analgesia. PMID- 10834091 TI - [The characteristics of the inotropic effects of Levosimendan on the isolated myocardium of patients with ischemic heart disease]. AB - Effect of the new cardiotonic agent levosimendan on the inotropic response parameters of the isolated myocardium of patients suffering from ischemic heart disease (IHD) was studied in the 0.01-1.00 mumole/liter concentration range. The inotropic response was measured in an isometric mode (electric stimulation frequency, 0.5 Hz; temperature, 37 degrees C; perfusion rate, 10 ml/min). In the range of 0.01-0.1 mumole/liter, levosimendan preferentially increases the rate of stress growth (+dP/dt) and the relaxation time. At concentrations above 0.1 mumole/liter, levosimendan produces a pronounced increase (56%) in the amplitude of single contraction. The mechanogram exhibits the signs of Ca-overload, including the post-contraction wave. It is concluded that the inotropic action of levosimendan on the isolated myocardium of IHD patients is related predominantly to increasing calcium uptake from outside. This circumstance is apparently caused by the initially high calcium content in sarcoplasmic reticulum, which is an important feature of the entire intracellular homeostasis in cardiomyocytes during IHD. PMID- 10834089 TI - [The evaluation of the specific activity and side effects of the m-cholinergic blocker pentifin compared to other antiparkinson agents]. AB - Experiments on rodents showed that pentifin, a muscarine antagonist belonging to the group of acetylene amines, possesses a pronounced antiparkinsonian activity. Pentifin is superior in the breadth of therapeutic action and tolerance characteristics to the conventional agents used for Parkinson's disease treatment. PMID- 10834092 TI - [The anti-arrhythmia and pro-arrhythmia properties of sigma-receptor ligands]. AB - It has been found that the sigma 1- and sigma 3-receptor antagonists, DuP 734 intraperitoneally and XJ 448 intravenously, had antiarrhythmic effects against epinephrine-induced arrhythmias in rats. The ED50 for antiarrhythmic effect of DuP 734 was 0.157 mg/kg after intraperitoneal administration. Other sigma receptor antagonists (rimcazole, BMY 14802, haloperidol) did not affect the incidence of epinephrine-induced arrhythmias. sigma 1-Receptor agonists (DTG, N allyl-normetazocine, (+)-3-PPP, (-)-3-PPP) had proarrhythmic effect after systemic administration. The sigma-agonist N-allyl-normetazocine and sigma antagonist XJ 488 had no effect on the incidence of epinephrine-induced arrhythmias after intracerebroventricular administration. Therefore, it appears that the central nervous system does not play a significant role in the antiarrhythmic or proarrhythmic effects of sigma ligands. We hypothesize that these effects of sigma ligands might be dependent on their action on cardiac sigma receptors. PMID- 10834090 TI - [A disorder of myocardial contractile function in acute experimental coronary failure: the submolecular mechanisms and the action of cardiac glycosides]. AB - Skinned and hybrid myocardial fibers were studied by methods of tensometry, determination of the ATP hydrolysis intensity, and resonance fluorescent energy transfer between highly selective labels bound to various amino acid residues. It was established that development of the early stage of heart failure in the case of acute myocardial ischemia caused by 15-min coronary artery occlusion (CAO) is related to a reversible damage or adaptive (functional) depression of the contractile protein system. As a result, the system features isolated submolecular post-translational variation in the properties of major proteins in a thin actin filament (myosin is not significantly damaged). This leads to a decrease in the force developed by the hybrid fibers (reconstructed using ghost myocardial fibers taken from ischemic area and normal myosin) and in the ATPase activity of actomyosin (ATP hydrolysis intensity) without any significant change in the Ca-sensitivity, cooperativity of the Ca-response of the actomyosin ensemble, and efficiency of the contractile process. In actin of the ischemic area, CAO results in a serious damage of the Lys61 and Cys374 regions and in a less pronounced damage of the Tyr69 and Cys10 regions. These results suggest that the Lys61 and, probably, Cys374-Lys61 regions are included in the actin monomer as a protomer, without adequate prepolymerization structural-conformational changes necessary to provide for the normal functioning of the filament. In the CAO-induced early stage of heart failure, cardiac glycosides (beta-acetyldigoxin, beta-methyldigoxin, and strophanthin K) produce a direct effect upon the intramolecular structure of myocardial actin, restore the generated force level, and increase the intensity of ATP hydrolysis by actomyosin ensemble. This is achieved by improving or normalizing the structural-conformational state and conformational mobility of the Lys61 and Cys374 region of actin. PMID- 10834093 TI - [A dried aspen bark extract in the experimental therapy of gastric peptic ulcer]. AB - Dry aspen bark extract exhibits pronounced antiulcerogenic activity as demonstrated by the results of experiments using the models of histamine and atophan induced ulcers and the ulceration model according to H. Shay. The gastroprotector activity is manifested by decreasing number of animals with ulceration damage in the mucous membrane of the stomach and markedly decreasing ulcer density. The antiulcerogenic effect of the dry aspen bar extract is comparable to that of the reference drugs alanton and gastrozepin. PMID- 10834094 TI - [The effect of reaferon on the level of catecholamines and mitochondrial enzymes in the ischemic kidney]. AB - The effect of reaferon (introduced at a dose of 1 x 10(6) IU/kg over six days prior to ischemia induction) on the levels of catecholamines and the activity of succinate dehydrogenase and NADH-dehydrogenase in various structures of kidney was studied in experiments on white rats. The ischemia was modeled by 90-min ligation of renal vessels. Reaferon retained the luminescence of catecholamines in all renal structures 24 h after circulation was restored on the level of intact kidney, except for the nerve trunks where the luminescence intensity decreased by 40%. Preliminary introduction of reaferon stimulated restoration of the enzymatic activity in the Krebs cycle and mitochondrial respiratory chain after 24- and 48-h revascularization, respectively. PMID- 10834095 TI - [An analysis of 3H-tamoxifen binding with the cell plasma membranes of the rat uterus]. AB - The effect of phencarol, ranitidine, propranolol, atropine, and diethylstilbestrol (known as modulators of the tamoxifen-binding sites in plasma membranes) on the 3H-tamoxifen binding to the plasma membrane fraction of white rat uterine cells was studied by determining the amounts of estradiol and H1- and H2-receptors. The interaction of tamoxifen with various receptors of uterine cells may be significant for evaluation of the tumor activity of this compound. PMID- 10834096 TI - [The hemorheological effects of Lychnis chalcedonica L. extracts]. AB - The hemorrheological effects of extracts from Maltese cross campion (Lychnis chalcedonica L.) prepared using 40% (I) and 70% (II) aqueous ethanol solutions were studied in vitro in comparison with tanakan on the model of high blood viscosity syndrome. Under the hyperthermia conditions, extracts I and II at a concentration of 1 x 10(-5) g/ml limited the growth of blood viscosity and thrombocyte aggregation and the drop in thrombocyte deformability. The effects of both extracts were comparable to those of tanakan. Prolonged (5 days) administration of extract I (daily dose, 150 mg/kg p.o) in rats with arterial hypertension (SHR) decreased the viscosity of whole blood and plasma, reduced the concentration of fibrinogen, and increased the deformability of erythrocytes. No reliable effects of the extracts in vivo upon the erythrocyte aggregation and hematocrit were observed. PMID- 10834097 TI - [The antimutagenic activity of afobazol studied in vivo]. AB - The chromosome aberration assay in the bone marrow cells of C57B1/6 mice showed that the new 2-mercaptobenzimidazole derivative afobazole(1-100 mg/kg) prevented manifestations of the clastogenic effects of dioxidine (100 mg/kg, i.p.) over a period of 24 h and reduced by 44-75% the cytogenetic effect of dioxidine (300 mg/kg, i.p.). The same doses of afobazole produced a statistically significant decrease in the cyclophosphamide (20 mg/kg, i.p.) damage over a period of 24 h. Afobazole showed no inherent mutagen activity and did not potentiate the effects of mutagens studied. PMID- 10834098 TI - [The use of the fluorimetric method for studying DNA integrity exemplified by an assessment of dioxidine genotoxicity]. AB - The genotoxic effect of dioxidine (300, 100, 30, and 10 mg/kg, i.p.) on the liver cells of outbred male rats was studied by fluorometric analysis of DNA unwinding (FADU) using the cell DNA samples taken from animals sacrificed 1, 6, 18, and 24 h after drug introduction. At a dose of 10 mg/kg, dioxidine produced no statistically significant increase in the amount of DNA damage relative to control. Increase in the dioxidine dose to 30 mg/kg resulted in a significant DNA damage 1 h after drug administration, while a dose of 100 or 300 mg/kg reliably caused damage at all times of exposure. PMID- 10834099 TI - [The dynamics of antigen-antibody complex formation in the presence of iodine containing diagnostic agents]. AB - A mathematical model is proposed to describe the antigen-antibody complex formation under the immunoassay conditions, which allows the antibody-antigen affinity to be evaluated. In particular, it is established that the dissociation constant of the antigen-antibody complex (for antibodies to a tuberculous Mycobacterium) confidently increases (p > 0.95) in the following sequence of diagnostic kits: omnipak > Ultravist > melitrast > Triombrast. The method suggested for analysis of the influence of xenobiotics upon the antigen-antibody reaction can be used to reveal their possible side effects on the antigen antibody complex formation in any particular case. PMID- 10834100 TI - [The effect of perftoran on the biochemical detoxication parameters in rats with an acute toxic lesion of the liver]. AB - Experiments on rats with heliotrine-induced acute liver damage showed the main parameters of biochemical detoxication to be inhibited. Treatment of the test animals with perftoran increased the initially low level of microsomal hemoproteins, the activity of enzymes participating at the second stage of biotransformation, the content of reduced glutathione, and the activity of enzymes involved in the glutathione metabolism. Perftoran decreased the level of lipid peroxidation in the microsomal fraction of damaged hepatocytes and activated enzymes involved in the antioxidant protector system. PMID- 10834101 TI - [A short essay on the history of the Department of Pharmacology of the Military Medical Academy over 200 years (1798-1998)]. PMID- 10834102 TI - [Reye's syndrome--a severe complication of salicylate therapy]. AB - Clinical and experimental data are reviewed on the Reye's syndrome--a heavy complication accompanying the therapy of viral infections in children by salicylates. Disorders in the bioenergetics of fatty acid oxidation and ammonia utilization are considered in the context of clinical manifestations of the Reye's syndrome. PMID- 10834103 TI - Therapeutic liver repopulation for the treatment of metabolic liver diseases. AB - Orthotopic liver transplantation is the treatment of choice for many inherited and acquired liver diseases. Unfortunately, the supply of donor organs is limiting and therefore many patients cannot benefit from this therapy. In contrast, hepatocyte suspensions can be isolated from a single donor liver can be transplanted into several hosts, and this procedure may help overcome the shortage in donor livers. In classic hepatocyte transplantation, however, only 1% of the liver mass or less can be replaced by donor cells. Recently though, we have used a mouse model of hereditary tyrosinemia to show that > 90% of host hepatocytes can be replaced by a small number of transplanted donor cells in a process we term "therapeutic liver repopulation". This phenomenon is analogous to repopulation of the hematopoietic system after bone marrow transplantation. Liver repopulation occurs when transplanted cells have a growth advantage in the setting of damage to recipient liver cells. Here we will review the current knowledge of this process and discuss the hopeful implications for treatment of liver diseases. PMID- 10834104 TI - Roles of intestinal trefoil factor (ITF) in human colorectal cancer: ITF suppresses the growth of colorectal carcinoma cells. AB - Intestinal trefoil factor (ITF) is a member of trefoil peptide family and is expressed almost exclusively in the goblet cells of small intestine and colon. Its expression is up-regulated by inflammatory and ulcerative conditions in the intestinal mucosa, and ITF has a role to maintain the mucosal integrity and repair the damaged mucosa. On the other hand, human colorectal carcinoma cells also express ITF peptide. In this review, we discussed the current views on the biological functions of ITF in the intestinal mucosa, and its suppressive effect on the growth of colorectal carcinoma cells. PMID- 10834105 TI - [Dietary fiber inhibits the incidence of hepatic metastasis with the anti-oxidant activity and portal scavenging functions]. AB - Dietary fiber is described as the proportion of plant foods not digested in the human small intestine. Among the various kinds of pectin, apple pectin exerts a bacteriostatic action and therefore may change the composition of the intestinal flora. The diet supplemented with 20% apple pectin significantly decreased the number and the incidence of AOM-induced colon tumors in rats. The prostaglandin E2 (PGE2) level in the distal colonic mucosa and blood of portal vein was lower in rats fed 20% pectin than those fed the basal diet. The ability of apple pectin to decrease PGE2 was dose-dependent and those results suggest an anti inflammatory effect in the bowel. Rats fed apple pectin showed a significantly lower incidence of hepatic metastasis than those fed the basal diet. To determine whether the anti-inflammatory effect of Lactobacillus on hepatic metastasis model same as apple pectin, Lactobacillus casei was selected. Metastatic nodules were significantly reduced, especially in the group receiving pretreatment. Apple pectic oligosaccharides with small molecular weights showed highly inhibitory effects on hypoxanthin-xanthin oxidase and ferrous sulfate-hydrogen peroxide. The scavenging activity of apple pectic oligosaccharides was suitable and accelerated at high temperatures (120 degrees C, 30 min.). Apple pectin and Lactobacillus have a scavenger effect in the intestinal digestion and portal circulation system as an anti-inflammatory food and have very important function for the prevention of hepatic metastasis. PMID- 10834106 TI - Model for mediastinal lymph node metastasis produced by orthotopic intrapulmonary implantation of lung cancer cells in mice. AB - This study is designed to establish a pulmonary tumor model to investigate the biology and therapy of lung cancer in mice. Current methods for forming a solitary intrapulmonary nodule and subsequent metastasis to mediastinal lymph nodes are not well defined. Lewis lung carcinoma cell (LLC) suspensions were orthotopically introduced into the lung parenchyma of C57/BL6 mice via a limited skin incision without thoracotomy followed by direct puncture through the intercostal space. The implantation process was performed within approximately 50 sec per mouse, and the operative mortality was less than 5%. Single pulmonary nodules developed at the implanted site in 93% of animals and subsequent mediastinal lymph nodes metastasis were observed in all mice that were succeeded to form a lung nodule after intrapulmonary implantation. The size of tumor nodule and the weight of mediastinal lymph node increased in a time-dependent manner. The mean survival time of mice implanted successfully with LLC cells was 21 +/- 2 days (range; 19-24 days). Histopathological analysis revealed that no metastatic tumor was detectable in the mediastinal lymph nodes on day 11, but metastatic foci at mediastinal lymph nodes were clearly observed on days 17 and 21 after implantation. Other metastases in distant organs or lymph nodes were not observed at 21 days after the implantation. Comparative studies with intrapleural and intravenous injections of LLC cells suggest that the mediastinal lymph node metastasis by intrapulmonary implantation is due to the release of tumor cells from the primary nodule, and not due to extrapulmonary leakage of cells. An intravenous administration of CDDP on day 1 after tumor implantation tended to suppress the primary tumor nodule and significantly inhibited the lymph node metastasis. Thus, a solitary pulmonary tumor nodule model with lymph node metastasis approximates clinical lung cancer, and may provide a useful basis for lung cancer research. PMID- 10834107 TI - [Effect of bufalin on growth and differentiation of human skin carcinoma cells in vitro]. AB - Bufalin, a cardiotonic steroid isolated from the Chinese toad, was previously shown to have growth inhibitory and differentiation inducing activities on leukemia cells and malignant melanoma cells. We examined the effect of bufalin on growth and differentiation of human skin squamous cell carcinoma cells (SSCC-1) in vitro. The concentration needed for growth inhibition of SSCC-1 cells was 10( 8) M, which was lower than those of gamabufotalin and ouabain. When SSCC-1 cells were treated with 10(-8) M bufalin for 16 h, the DNA synthesis of SSCC-1 cells decreased, but there was no change in their survival ratio. The results suggest that growth inhibitory effect of buffalin is not only a cytotoxic effect. Bufalin increased the production of cornified envelopes and the expression of Keratin K10/11 and involurcin. These findings indicate that bufalin has both growth inhibitory and differentiation inducing effects on SSCC-1 cells. PMID- 10834108 TI - Establishment and characterization of human uterine poorly differentiated adenocarcinoma cell line (HHUABM). AB - The cell line designated HHUABM was established from the metastatic region (left Bartholin gland) of human endometrial adenocarcinoma. The cell line grew well, multilayering rapidly without contact inhibition, and 72 serial passages were successively done within 25 months. The cultured cells of HHUABM line were round and spindle in shape, and showed a pavement-like arrangement. The distribution of chromosome number varied narrowly at the diploid range, and the modal chromosome number was 46. The 90% of metaphase cells showed normal karyotype. The HHUABM cells were transplanted easily into the subcutis of BALB/c nude mice and produced poorly differentiated adenocarcinoma resembling the original tumor. The conditioned medium promoted the proliferation of CPAE (endothelial cells). The estradiol-17 beta and progesterone receptors were not detected. PMID- 10834109 TI - Establishment and characterization of human choriocarcinoma cell line derived from a metastatic focus of a testicular mixed germ cell tumor. AB - A human testicular choriocarcinoma cell line HKRT-II was established by the single-cell cloning method from a mixed cell culture system derived from a retroperitoneal metastatic germ cell tumor composed of a yolk-sac tumor, a choriocarcinoma, and an immature teratoma. Its primary tumor rose from the testis and was comprised of a seminoma, a yolk-sac tumor, a choriocarcinoma and an immature teratoma. The HKRT-II cells were spindle or polygonal in shape and contained multi-nucleated giant cells showing neoplasticity and pleomorphism. The cells proliferated in a stable manner, and the population doubling time was 42 hours. The chromosome numbers showed a wide distribution of aneuploidy, while the mode was in the hypertetraploid range. Double minute chromosomes and homogeneously staining regions were recognized in about 5% to 10% of the metaphase plates, respectively. Heterotransplantation was not difficult. Subcutaneous transplantation of 1 x 10(7) cells into nude mice formed a tumor composed of only a choriocarcinoma. The most noteworthy characteristics of the cell line were that it produced human chorionic gonadotropin (hCG) in an in vitro culture system and in in vivo grafted cells, and that the N-myc gene was amplified about 10 times. PMID- 10834110 TI - Detection of mycoplasma contaminations in cell cultures by PCR analysis. AB - Mycoplasma contamination is still one of the main problems in using cell cultures in biological and medical research and in the production of bioactive substances, because mycoplasma can alter nearly all parameters and products of the cell. They can persist undetected in the culture if no special detection methods are applied. In recent years, the PCR technology has become a commonly used method to analyze genomic DNA and the expression of genes, with both high specificity and sensitivity. This technique can be effectively employed for the detection and even the identification of mycoplasma contaminations in cell cultures applying primers complementary to the 16S rDNA region. Although this technique, once established, is characterized by simplicity and speed, PCR is still a complex process and its sensitivity and specificity can be influenced by a number of different parameters, e.g. inhibiting compounds originating from the preparation process of the DNA, RNA or cDNA, contamination of the solutions with PCR products, and the selection of a primer pair which does not cover all the mycoplasma species occurring in cell cultures. Thus, adequate controls have to be included to obtain reliable results. The present review examines the use of different primers of the 16S rDNA region including their specificity, the sensitivity applying various DNA or RNA preparation procedures, and the methods to detect finally the amplicons. In conclusion, basic nucleic acid preparation and PCR product detection methods offer a simple, fast and reliable technique for the examination of mycoplasma contaminations in cell cultures, provided that the indispensable control assays are implemented. PMID- 10834111 TI - [Brain tumors and chemokines]. PMID- 10834113 TI - The growing applications of medical infrared imaging. PMID- 10834112 TI - Information from motion. PMID- 10834114 TI - Functional infrared imaging of the breast. PMID- 10834115 TI - Applying dynamic thermography in the diagnosis of breast cancer. PMID- 10834116 TI - The important role of infrared imaging in breast cancer. PMID- 10834117 TI - The potential of dynamic area telethermometry in assessing breast cancer. PMID- 10834118 TI - Increasing the effective resolution of thermal infrared images. PMID- 10834119 TI - Converting far infrared image information to other physiological data. PMID- 10834120 TI - Cardiothermographic assessment of arterial and venous revascularization. PMID- 10834121 TI - Thermographic evaluation of myocardial protection. PMID- 10834123 TI - An automated approach to beat-by-beat QT-interval analysis. PMID- 10834122 TI - Independent component analysis for EEG source localization. PMID- 10834124 TI - Studying the significance of cardiac contractility variability. PMID- 10834125 TI - The importance of pulsatile microcirculation in relation to hypertension. PMID- 10834126 TI - The frontier of substitution medicine: integrating biomaterials and tissue engineering. PMID- 10834127 TI - [Guidelines: some considerations on one of the cultural instruments on which the switch in the National Health System can be based]. AB - In the last few years there has been an explosion of interest in guidelines, reflected in a vivid debate in the international literature. Guidelines for medical practice are not innovative tools, since their history began at least 20 years ago. They are produced with increasing frequency in North America, and similar developments are taking place in Europe. The concept of improving the quality of the services in the National Health System through the production and implementation of valid clinical guidelines has been recognized in Italy by the Ministry of Health, who introduced such a concept in the last reform of the law on the Sanitary Health System. Many international authors deem that clinical guidelines are a way to support effective clinical practice. If this still holds, and we believe it does, then we must ensure that these guidelines are effective. Also for the enthusiast, the entire process from the development to the dissemination, implementation, and evaluation is not an easy task. Guidelines should be written correctly by a multidisciplinary panel of experts, should be peer reviewed and updated. Guidelines should become the common field in which the four major parties of the sanitary "market" (a market far from ideal today) meet: the patient, the doctor, the manager, and the industry involved in medical technology. Guidelines should be disseminated and implemented: simply having evidence available does not necessarily mean that it will be used. Thus, the key question is not so much related to the acceptance of best practice, but the extent to which a guideline is cost-effective. In this review we have tried to discuss such points, taking into consideration all the steps, the discussions, the controversial points, and the unsolved questions which are smoldering under both the potential benefit of practice guidelines (also in the light of appropriateness of medical practice), and all the possible effects on clinical autonomy, health care costs, clinical practice satisfaction, legal implications, and conflicts of interest. It is crucial for medical associations, and indeed for all physicians, to take up this problem and constructively try to play an active part in the process, avoiding the possibility that controversial guidelines be imposed by the regulatory authorities. PMID- 10834128 TI - [The electrocardiology of atrial fibrillation]. AB - Atrial fibrillation (AF) is the most common sustained cardiac arrhythmia and it is associated mainly with a significant mortality and morbidity. The purpose of this report is to focus on the major electro-cardiological aspects of AF and to provide some elements useful in the understanding of its pathophysiology. Experimental and clinical studies have shown that AF is maintained by multiple reentrant wavelets within the atrial muscle. It has been estimated that a critical number of wavelets (from 3 to 6) is necessary for perpetuation of AF. The short duration of the atrial effective refractory period (ERP) usually favors the onset of AF. Two different kinds of reentry have been observed during AF: random reentry and leading circle reentry. The presence of an adequate substrate is critical for the beginning of AF. Although this event is possible even in a normal atrium, atrial dilation and/or structural heterogeneity enhance the atrial propensity to develop AF. Even uniform or non-uniform anisotropy is now believed to play an important role in the genesis of AF. Finally, ionic channel disorders, some of which are genetically transmitted, may represent a possible substrate of AF. An important modulating role of the autonomic nervous system in the genesis of AF is universally accepted. In particular, many observations support the hypothesis that patients without heart disease tend to have vagally-mediated AF, whereas patients with structural heart disease tend to have adrenergic-mediated AF. In the presence of an opportunely modulated substrate, a third prerequisite for the triggering of a multiple atrial reentry is the presence of an adequate "trigger" factor. This is represented, in most cases, by ectopic atrial beats, commonly originating in the pulmonary veins. AF may cause atrial changes, either in an electrophysiological behavior and anatomy or both, that may favor its irreversibility and/or its frequent recurrence. Some of these changes are: atrial ERP disease, paradoxical shortening of ERP at a lower rate (inversed rate adaptation), accumulation of glycogen within atrial cells, apoptosis, and cellular dedifferentiation. The ventricular rate during AF has a pivotal role in its pathophysiology. The ERP of the atrioventricular node, the concealed conduction through the atrioventricular node, the autonomic neural balance and the drug's action are the most important factors that regulate ventricular rate during AF. In the presence of an atrioventricular accessory pathway, with fast anterograde conduction, AF is a very dangerous arrhythmia. During AF, aberrant conduction may often be seen, especially after long cycles (Ashman phenomenon). In patients with dilated cardiomyopathy and AF, arrhythmia is not always a secondary phenomenon. In fact, there is current evidence that AF may be the cause of cardiomyopathy. PMID- 10834129 TI - [The assessment of breathing during sleep: a curiosity or clinical necessity?]. AB - The study of sleep, which initially focused on the neurophysiological mechanisms and cardiorespiratory function during the night, has shown the presence of sleep related breathing disorders that epidemiological, pathophysiological and clinical data have indicated to be associated with increased cardiovascular morbidity and mortality: the obstructive sleep apnea syndrome (OSAS) and the central sleep apnea syndrome (CSAS). OSAS is a condition characterized by repetitive respiratory pauses due to the pharynx wall collapse, with a subsequent obstruction to the airflow. The hemodynamic consequences due to the markedly increased negative intrathoracic pressure (induced by the respiratory muscle effort towards the closed upper airways), the progressive hypercapnic hypoxemia and the arousal terminating the apneas, are the pathophysiological keys of the cardiovascular effects of OSAS and may explain the association between OSAS and the documented increase of cardiovascular morbidity and mortality. CSAS is a breathing disorder characterized by recurrent episodes of central hypopneas or apneas and hyperventilation which, is the classical form described by Cheyne and Stokes, show a crescendo-decrescendo pattern of respiration. Pathophysiological and epidemiological data clearly indicate the link between CSAS and heart failure, also showing a correlation between respiratory disorders and the severity of hemodynamic impairment. However, other mechanisms are involved in the genesis of CSAS in explaining the variable presence of CSAS independent of cardiac function and, more importantly, the impact of CSAS on poor prognosis in heart failure. In conclusion, the data available indicate the need to include screening for sleep-related breathing disorders in the evaluation of cardiac patients who are at risk for OSAS and, particularly, in patients with heart failure, who could really benefit from treatment of the respiratory disorder. PMID- 10834130 TI - [The appropriateness of hospital admissions: reflections for a global approach]. AB - In accordance with a recent definition, appropriateness of medical care is a diagnostic-therapeutic intervention with a correct use of resources and competency, and a procedure that satisfies the request of patients. This definition considers not only the risk/benefit relationship but also the interaction between costs and benefits. In this light, the concept of appropriateness needs a global approach. The identification of the Mission and Vision of the structure, the use of medical guidelines, the distribution of documents which inform patients on the procedures carried out in the hospital and the modalities of access to the structure, the implementation of procedures which define hospital admission and finally the random check of medical records could represent important steps to implement a quality system aimed at guaranteeing the appropriateness of hospital admissions in accordance with the international standards for total quality management ISO 9000. PMID- 10834131 TI - [Bereavement. The management of mourning: the cardiovascular physiopathology of communication]. AB - Within the communication competence requested to an experienced physician, bereavement has a relevant role. A case of a patient with postinfarction cardiogenic shock admitted to a cardiosurgery department for rescue myocardial revascularization is described; the patient's wife, after communication of the death of her husband during the surgical procedure, developed an acute anterior myocardial infarction with angiographic normal coronary arteries. Pathophysiological issues concerning acute ischemic event triggers are discussed, with particular attention to emotional stress subtypes, their neuronal loops and their interactions with different cardiovascular diseases (arrhythmic, congenital, atherosclerotic). The essential variables of an adequate bereavement are discussed: set, mood, language, cultural background. The sympatho-vagal unbalance induced by emotional stress as an acute ischemic event trigger is emphasized as well as the need for more awareness by health care operators of the emotional impact and adequate management of bereavement. PMID- 10834132 TI - [The assessment of the coronary risk in a work setting. The results of the SEMM study and the outlook for prevention in Italy. Surveillance of Employees Municipality of Milan]. AB - BACKGROUND: The goals of the present report are to assess the differences in distribution of traditional coronary risk factors in a work setting, aimed at identifying specific groups at risk and to compare mean values of such risk factors and of an overall risk score of the entire working sample with the results observed in the third WHO-MONICA population survey carried out in northern Italy (Area Brianza). METHODS: In the SEMM study 7872 employees (2601 men and 5271 women) were enrolled between 1992 and 1996. The third MONICA survey in Brianza was carried out on an age- and gender-stratified random sample of 831 men and 884 women in 1993-1994, selected from the 25-64-year-old residents of five municipalities, representative of the study population. In both studies coronary risk factors were measured according to the MONICA protocol, adopting standardized methods. RESULTS: In comparison with the MONICA population sample, the entire working group showed lower mean levels of blood pressure and total cholesterol, higher prevalence of current cigarette smokers and lower mean levels of HDL cholesterol, in both gender groups. Prevalence of overweight subjects was higher among men in the working group, but the opposite pattern was detected in women. The overall risk score, calculated using the coefficients of a proportional hazard survival equation estimated in a large collaborative Italian follow-up study, resulted lower in the working sample, in both gender groups. This result may be attributed to a selection bias known in occupational epidemiology as "healthy worker effect". In contrast to this finding, the prevalence of smokers, in particular among women, was higher in the employed sample, indicating that working stress conditions may play some role. CONCLUSIONS: In order to extend the assessment of cardiovascular risk factors as well as prevention activities in work settings, some advantages are highlighted: the high participation rates, the feasibility to adopt standardized protocols, and easier and cheap procedures for censoring in follow-up studies. Moreover, due to the recently adopted legislation in Italy which increases the number of working categories to be included in periodic clinical examinations, prevention activities in work settings to contrast the epidemic of widespread chronic diseases, like cardiovascular diseases, are encouraged. This will also allow for the investigation of individual variations over time of coronary risk factors. PMID- 10834133 TI - [A reduction in the invasiveness during surgical revascularization: the harvesting of the great saphenous vein by a video endoscopic technic]. AB - BACKGROUND: The authors report their preliminary experience of endoscopic saphenous vein harvesting as part of a program devoted to reducing the invasivity of surgical myocardial revascularization. This method allows us to minimize the cutaneous incisions in the inferior limbs necessary to harvest the saphenous vein, thus reducing the incidence of complications. METHODS: The study includes 41 patients who underwent endoscopic saphenous vein harvesting from October 1998 to September 1999 and, as a control group, 20 patients with similar characteristics operated on with the traditional technique during the same period. The variables considered were: the time necessary to harvest the saphenous vein, the incidence of complications, and the postoperative mobilization. RESULTS: All the endoscopically harvested grafts were adequate for the scheduled procedure. The only complication occurred in a patient operated on with the traditional technique. The time of harvesting and the day of mobilization were similar in the two groups. The reduction of surgical trauma allowed a fast deambulation recovery and better esthetic results. CONCLUSIONS: When complete arterial revascularization is not feasible, the endoscopic harvesting of the required saphenous vein segment allows for a significant reduction in the invasivity of the procedure. PMID- 10834134 TI - [Vasoactive intestinal peptide in heart failure]. AB - BACKGROUND: The aim of our study was to investigate the pathophysiological role of the vasoactive intestinal peptide (VIP), a vasodilating neuropeptide with positive inotropic and chronotropic properties, in heart failure. METHODS: The study was carried out in 35 patients with heart failure due to dilated cardiomyopathy, who underwent a peripheral venous blood sample for radioimmunoassay of VIP within the first in-hospital day. RESULTS: The plasma concentration of VIP: 1) is not higher than normal in the whole group of patients with heart failure; 2) is higher in younger than in elderly healthy subjects but does not significantly change in relation to age in heart disease patients; 3) is higher in elderly (> 60 years) but not in younger (< 60 years) patients compared to healthy subjects of the same age; 4) is higher in NYHA functional class 2 than in NYHA functional class > 2 groups and in normal subjects; 5) is not correlated with echocardiographic parameters; 6) does not significantly change with respect to the etiology of dilated cardiomyopathy. CONCLUSIONS: The plasma concentration of VIP in heart failure is conditioned by some epidemiological and clinical variables. Unlike the healthy group, differences are not detectable with respect to the age of patients; thus, in elderly heart disease subjects the neuropeptide productive potentiality is preserved. Taking into account the physiological properties of VIP, its plasma increase in the initial phase of heart failure can be reasonably regarded as a further mechanism to restore the compromised hemodynamic balance. Its decrease, related to worse clinical conditions, could be due to a progressive depletion from the pre-synaptic nerve endings and to a deficiency in the neurogenic productive capacity of the molecule. PMID- 10834135 TI - [Acute myocardial damage from a pheochromocytoma]. AB - Cardiac involvement in pheochromocytoma is rare but may be associated with serious clinical deterioration. A 70-year-old woman arrived at our Emergency Department because of chest discomfort, blood pressure lability, mild dyspnea and electrocardiographic signs suggesting an acute myocardial infarction. However two dimensional echocardiogram did not show any segmental wall motion abnormalities but diffuse and severe left ventricular hypokinesia. The patient was treated with ACE-inhibitors and diuretics and did not receive thrombolytics or beta blocking agents. Creatine kinase-MB and troponin I were normal. Electrocardiogram and echocardiogram completely returned to normal within 1 week and a coronary angiography demonstrated normal coronary arteries. An increase in the catecholamine concentration in a 24-hour urinary sample suggested a pheochromocytoma that was confirmed by abdominal computerized tomography. During surgery, marked hypertension developed treated with sodium nitroprusside and labetalol, and after removal of the tumor severe hypotension required infusion of norepinephrine for several days. PMID- 10834136 TI - [A synthesis of the guidelines of the ESC/EAS/ESH on the prevention of coronary disease and of the guidelines of the Task Force for the Prevention of Coronary Heart Disease. European Society of Cardiology]. PMID- 10834137 TI - [The prevention of coronary disease in clinical practice. The recommendations of the Second Joint Task Force of European Societies on the Prevention of Coronary Pathology. Societa Europea di Cardiologia, Societa Europea dell'Aterosclerosi, Societa Europea dell'Ipertensione]. PMID- 10834138 TI - [In Process Citation] AB - Traduzione integrata del documento redatto dall'International Task Force for Prevention of Coronary Heart Disease in collaborazione con l'International Atherosclerosis Society dal titolo "Coronary heart disease: reducing the risk. The scientific background for primary and secondary prevention of coronary heart disease. A worldwide view" pubblicato in extenso in Nutrition Metabolism Cardiovascular Diseases (1998; 8: 205-71, Assmann G, Carmena R, Cullen P, Fruchart JC, Lewis B, Mancini M, Olsson A, Paoletti R, Pometta D, Tikkanen M) ed in forma ridotta su Circulation (1999; 100: 1930-8, Assmann G, Carmena R, Cullen P, Fruchart JC, Jossa F, Lewis B, Mancini M, Paoletti R, for the International Task Force for Prevention of Coronary Heart Disease. Coronary heart disease: reducing the risk. A worldwide view) e su Arteriosclerosis, Thrombosis, and Vascular Biology (1999; 19: 1819-24, Assmann G, Cullen P, Jossa F, Lewis B, Mancini M, for the International Task Force for the Prevention of Coronary Heart Disease. Coronary heart disease: reducing the risk. The scientific background to primary and secondary prevention of coronary heart disease). PMID- 10834139 TI - [The usefulness of CPK-MB and troponin I in the management of chest pain in first aid]. PMID- 10834140 TI - [Warfarin anticoagulation in patients with nonvalvular atrial fibrillation]. PMID- 10834141 TI - [Angioplasty with or without a stent in acute infarct]. PMID- 10834142 TI - [The reduction of cardiovascular risk with ramipril]. PMID- 10834143 TI - [The transcatheter ablation of ventricular tachycardia: a cost-effectiveness analysis]. PMID- 10834144 TI - [Integration between hospital and region: the case of heart failure]. PMID- 10834145 TI - [Reflections on health reform]. PMID- 10834146 TI - [Proposed mechanism of action of metalloendopeptidase-F in the treatment of patients with chronic hepatitis B or C infection]. AB - Chronic hepatitis B and C virus infections have been characterized by the pathophysiological features with a high incidence of progression to cirrhosis and development of hepatocellular carcinoma. The viral persistence produced by escape mutations from virus-specific cytotoxic T lymphocytes (CTL) response may lead to upregulation of delayed-type hypersensitivity immune response, which causes hepatic tissue damage through non specific macrophage activation and CTL response and promotes pathogenesis of hepatic fibrosis. In a preliminary clinical study, a novel metalloendopeptidase-F (MEP-F) has been shown to be effective in the treatment of patients with either chronic hepatitis B or C infection. Oral administration of MEP-F resulted in a significant reduction of the serum levels of HBs antigen and HCV RNA and improvement in the liver function abnormalities. However, the mechanism of action of MEP-F is not yet well understood. There are accumulating evidences showing an important role of alpha 2-macroglobulin proteinase complexes in regulatory mechanisms of immune response and repairing within impaired and inflammatory tissues. In this article, reviewing the pharmacological and biological properties of alpha 2-macroglobulin-proteinase complexes, the mechanism of anti-viral effect of MEP-F is examined based on the clinical findings. It is indicated that alpha 2-macroglobulin-MEP-F complexes may induce macrophage/Kuppfer cell activation and proliferation through binding their receptors on the cells and activating signaling cascades, which enhance both anti viral specific and nonspecific immune responses. alpha 2-Macroglobulin-MEP-F complexes may also augment cellular immunity and hepatic regeneration by neutralizing the immunosuppressive and fibrogenic activities of transforming growth factor-beta. PMID- 10834148 TI - [Evaluation of the activity and effects of combinations of various antibacterial agents against methicillin-resistant Staphylococcus aureus in vitro]. AB - MICs of various antibacterial agents against methicillin-resistant Staphylococcus aureus (MRSA) were measured. Furthermore, we evaluated the effects of combinations of antibacterial agents against MRSA in vitro. In 24 cases out of 37, in which MRSA was isolated from inpatients, other microorganisms, such as Candida spp., Entrococcus spp., and Pseudomonas aeruginosa, were simultaneously isolated. From the results of minimum inhibitory concentrations (MICs), obtained from micro broth-dilution method, of various antibacterial agents against MRSA, range of MICs of arbekacin (ABK), vancomycin (VCM) and teicoplanin (TEIC) were < or = 0.25-4.0, 0.5-1.0 and 0.25-4.0 micrograms/ml respectively, and no strains of MRSA showed resistance to ABK, VCM and TEIC, so that we concluded that these three antibacterial agents were effective for MRSA infection. On the in vitro study of combination-effect of antibacterial agents, significant synergistic effects were achieved in the combination of VCM and flomoxef (FMOX) (Synergism rate was 97.3%) or VCM and imipenem (IPM) (Synergism rate was 97.2%). From the results that the fractional inhibitory concentration index in the combination of VCM with IPM was smaller than that with FMOX and that P. aeruginosa or Enterococcus spp. were simultaneously isolated in high frequency in the MRSA isolated cases, we thought that the combination of VCM with IPM is more useful, because IPM is effective against P. aeruginosa but FMOX is not. PMID- 10834147 TI - [Changes in the antibacterial activity of chemotherapeutic agents (especially carbapenems) for 10 species of clinical isolates between 1994 and 1996. Surveillance group of the sensitivities of clinical isolates to antibacterial agents]. AB - During October and December of each year of from 1994 to 1996, 3,849 strains of 10 species of bacteria were isolated from clinical materials in 21 institutions nationwide. The minimum inhibitory concentrations (MICs) for these bacteria of four carbapenems (imipenem [IPM], panipenem [PAPM], meropenem [MEPM], and biapenem [BIPM]) and other representative antibacterial agents were measured to investigate annual changes in antibacterial activity. Carbapenems showed potent activity against methicillin-sensitive S. aureus (MSSA), S. pneumoniae, E. faecalis, H. influenzae, E. coli, K. pneumoniae, E. cloacae, S. marcescens, and the B. fragilis group, with the activity being stable. However, these drugs showed weak activity against methicillin-resistant S. aureus (MRSA) and P. aeruginosa. The antibacterial activity (MIC90) against the tested organisms generally remained stable. Particularly, there was annual improvement of the MIC90 values of IPM and BIPM for S. pneumoniae, as well as the values of IPM and PAPM for H. influenzae, and those of IPM, PAPM, and BIPM for S. marcescens. On the other hand, the activity of carbapenems (including IPM) against MRSA was not necessarily strong, but there was annual improvement of MIC90 values. PMID- 10834149 TI - [In vitro activity of faropenem against beta-lactamase producing clinical isolates]. AB - Each 20 strains of beta-lactamase producing methicillin susceptible Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Haemophilus influenzae, Moraxella (Branhamella) catarrhalis, and Bacteroides fragilis group were used as the test strains. Drug susceptibility of these strains to faropenem (FRPM), cefdinir, cefditoren, cefcapene, cefteram, cefaclor, and ampicillin was determined by an agar dilution method according to the NCCLS guideline M100-S9. beta-Lactamase activity of the test strains was determined by a spectrophotometric method. In the present study, FRPM was highly active against beta-lactamase-producing strains, and no close correlation was found between the MICs of FRPM for the test strains and their beta-lactamase activities. These results suggest that FRPM has potential in successful application for the treatment of infectious diseases with various types of bacterial pathogens including beta-lactamase producing strains. PMID- 10834150 TI - Three-dimensional computational model of left heart diastolic function with fluid structure interaction. AB - Aided by advancements in computer speed and modeling techniques, computational modeling of cardiac function has continued to develop over the past twenty years. The goal of the current study was to develop a computational model that provides blood-tissue interaction under physiologic flow conditions, and apply it to a thin-walled model of the left heart. To accomplish this goal, the Immersed Boundary Method was used to study the interaction of the tissue and blood in response to fluid forces and changes in tissue pathophysiology. The fluid mass and momentum conservation equations were solved using Patankar's Semi-Implicit Method for Pressure Linked Equations (SIMPLE). A left heart model was developed to examine diastolic function, and consisted of the left ventricle, left atrium, and pulmonary flow. The input functions for the model included the pulmonary driving pressure and time-dependent relationship for changes in chamber tissue properties during the simulation. The results obtained from the left heart model were compared to clinically observed diastolic flow conditions for validation. The inflow velocities through the mitral valve corresponded with clinical values (E-wave = 74.4 cm/s, A-wave = 43 cm/s, and E/A = 1.73). The pressure traces for the atrium and ventricle, and the appearance of the ventricular flow fields throughout filling, agreed with those observed in the heart. In addition, the atrial flow fields could be observed in this model and showed the conduit and pump functions that current theory suggests. The ability to examine atrial function in the present model is something not described previously in computational simulations of cardiac function. PMID- 10834151 TI - Turbulence characteristics downstream of bileaflet aortic valve prostheses. AB - This study was focused on a series of in vitro tests on the turbulent flow characteristics of three bileaflet aortic valves: St. Jude Medical (SJM), CarboMedics (CM), and Edwards Tekna (modified Duromedics, DM). The flow fields of the valves were measured in a pulsatile flow model with a laser-Doppler anemometer (LDA) at the aortic sinus area downstream of the valves. The heart rate was set at 70 beats per minute, the cardiac output was maintained at 5 liters per minute, and the aortic pressure wave forms were kept within the physiological range. Cycle-resolved analysis was applied to obtain turbulence data, including mean velocity, Reynolds stresses, autocorrelation coefficients, energy spectral density functions, and turbulence scales. The Reynolds shear stresses of all three valves induced only minor damage to red blood cells, but directly damaged the platelets, increasing the possibility of thrombosis. The smallest turbulence length scale, which offers a more reliable estimate of the effects of turbulence on blood cell damage, was three times the size of red blood cells and five times the size of platelets. This suggests that there is more direct interaction with the blood cells, thus causing more damage. PMID- 10834152 TI - Vortex shedding as a mechanism for free emboli formation in mechanical heart valves. AB - The high incidence of thromboembolic complications of mechanical heart valves (MHV) limits their success as permanent implants. The thrombogenicity of all MHV is primarily due to platelet activation by contact with foreign surfaces and by nonphysiological flow patterns. The latter include elevated flow stresses and regions of recirculation of blood that are induced by valve design characteristics. A numerical simulation of unsteady turbulent flow through a bileaflet MHV was conducted, using the Wilcox k-omega turbulence model for internal low-Reynolds-number flows, and compared to quantitative flow visualization performed in a pulse duplicator system using Digital Particle Image Velocimetry (DPIV). The wake of the valve leaflet during the deceleration phase revealed an intricate pattern of interacting shed vortices. Particle paths showed that platelets that were exposed to the highest flow stresses around the leaflets were entrapped within the shed vortices. Potentially activated, such platelets may tend to aggregate and form free emboli. Once formed, such free emboli would be convected downstream by the shed vortices, increasing the risk of systemic emboli. PMID- 10834153 TI - Helical flow around arterial bends for varying body mass. AB - The three dimensionally curved aortic arch is modeled as a portion of a helical pipe. Pulsatile blood flow therein is calculated assuming helical symmetry and an experimentally measured pressure pulse. Appropriate values for the Womersley and Reynolds numbers are taken from allometric scaling relations for a variety of body masses. The flow structure is discussed with particular reference to the wall shear, which is believed to be important in the inhibition of atheroma. It is found that nonplanar curvature limits the severity of flow separation at the inner bend, and reduces spatial variation of wall shear. PMID- 10834154 TI - Viscoelastic behavior of a lung alveolar duct model. AB - A study is conducted into the oscillatory behavior of a finite element model of an alveolar duct. Its load-bearing components consist of a network of elastin and collagen fibers and surface tension acting over the air-liquid interfaces. The tissue is simulated using a visco-elastic model involving nonlinear quasi-static stress-strain behavior combined with a reduced relaxation function. The surface tension force is simulated with a time- and area-dependent model of surfactant behavior. The model was used to simulate lung parenchyma under three surface tension cases: air-filled, liquid-filled, and lavaged with 3-dimenthyl siloxane, which has a constant surface tension of 16 dyn/cm. The dynamic elastance (Edyn) and tissue resistance (Rti) were computed for sinusoidal tidal volume oscillations over a range of frequencies from 0.16-2.0 Hz. A comparison of the variation of Edyn and Rti with frequency between the model and published experimental data showed good qualitative agreement. Little difference was found in the model between Rti for the air-filled and lavaged models; in contrast, published data revealed a significantly higher value of Rti in the lavaged lung. The absence of a significant increase in Rti for the lavaged model can be attributed to only minor changes in the individual fiber bundle resistances with changes in their configuration. The surface tension was found to make an important contribution to both Edyn and Rti in the air-filled duct model. It was also found to amplify any existing tissue dissipative properties, despite exhibiting none itself over the small tidal volume cycles examined. PMID- 10834155 TI - Aerosol transport and deposition in sequentially bifurcating airways. AB - Deposition patterns and efficiencies of a dilute suspension of inhaled particles in three-dimensional double bifurcating airway models for both in-plane and 90 deg out-of-plane configurations have been numerically simulated assuming steady, laminar, constant-property air flow with symmetry about the first bifurcation. Particle diameters of 3, 5, and 7 microns were used in the simulation, while the inlet Stokes and Reynolds numbers varied from 0.037 to 0.23 and 500 to 2000, respectively. Comparisons between these results and experimental data based on the same geometric configuration showed good agreement. The overall trend of the particle deposition efficiency, i.e., an exponential increase with Stokes number, was somewhat similar for all bifurcations. However, the deposition efficiency of the first bifurcation was always larger than that of the second bifurcation, while in general the particle efficiency of the out-of-plane configuration was larger than that of the in-plane configuration. The local deposition patterns consistently showed that the majority of the deposition occurred in the carinal region. The distribution pattern in the first bifurcation for both configurations were symmetric about the carina, which was a direct result of the uniaxial flow at the inlet. The deposition patterns about the second carina showed increased asymmetry due to highly nonuniform flow generated by the first bifurcation and were extremely sensitive to bifurcation orientation. Based on the deposition variations between bifurcation levels and orientations, the use of single bifurcation models was determined to be inadequate to resolve the complex fluid particle interactions that occur in multigenerational airways. PMID- 10834156 TI - Surfactant transport over airway liquid lining of nonuniform depth. AB - Numerous effects (e.g., airway wall buckling, gravity, airway curvature, capillary instabilities) give rise to nonuniformities in the depth of the liquid lining of peripheral lung airways. The effects of such thickness variations on the unsteady spreading of a surfactant monolayer along an airway are explored theoretically here. Flow-induced film deformations are shown to have only a modest influence on spreading rates, motivating the use of a simplified model in which the liquid-lining depth is prescribed and the monolayer concentration satisfies a spatially inhomogeneous nonlinear diffusion equation. Two generic situations are considered: spreading along a continuous annular liquid lining of nonuniform depth, and spreading along a rivulet that wets the airway wall with zero contact angle. In both cases, transverse averaging at large times yields a one-dimensional approximation of axial spreading that is valid for the majority of the monolayer. However, a localized monolayer remains persistently two dimensional in a region at its leading edge having axial length scales comparable to the length scale of transverse depth variation. It is also shown how the transverse spreading of a monolayer may be arrested as it approaches a static contact line at the edge of a rivulet. Implications for Surfactant Replacement Therapy are discussed. PMID- 10834157 TI - Viscoelastic dissipation in compact bone: implications for stress-induced fluid flow in bone. AB - Viscoelastic properties of wet and dry human compact bone were studied in torsion and in bending for both the longitudinal and transverse directions at frequencies from 5 mHz to 5 kHz in bending to more than 50 kHz in torsion. Two series of tests were done for different longitudinal and transverse specimens from a human tibia. Wet bone exhibited a larger viscoelastic damping tan delta (phase between stress and strain sinusoids) than dry bone over a broad range of frequency. All the results had in common a relative minimum in tan delta over a frequency range, 1 to 100 Hz, which is predominantly contained in normal activities. This behavior is inconsistent with an optimal "design" for bone as a shock absorber. There was no definitive damping peak in the range of frequencies explored, which could be attributed to fluid flow in the porosity of bone. PMID- 10834158 TI - A linear material model for fiber-induced anisotropy of the anulus fibrosus. AB - The anulus fibrosus (AF) is a lamellar, fibrocartilaginous component of the intervertebral disc, which exhibits highly anisotropic behaviors in tension. These behaviors arise from the material's unique collagen structure. We have investigated the use of a linear, fiber-induced anisotropic model for the AF using a quadratic strain energy density formulation with an explicit representation of the collagen fiber populations. We have proposed a representative set of intrinsic material properties using independent datasets of the AF from the literature and appropriate thermodynamic constraints. The model was validated by comparing predictions with previous experimental data for AF behavior and its dependence on fiber angle. The model predicts that compressible effects may exist for the AF, and suggests that physical effects of the equivalent "matrix," "fiber," "fiber-matrix," and "fiber-fiber," interactions may be important contributors to the mechanical behavior of the AF. PMID- 10834159 TI - A special theory of biphasic mixtures and experimental results for human annulus fibrosus tested in confined compression. AB - A finite deformation mixture theory is used to quantify the mechanical properties of the annulus fibrosus using experimental data obtained from a confined compression protocol. Certain constitutive assumptions are introduced to derive a special mixture of an elastic solid and an inviscid fluid, and the constraint of intrinsic incompressibility is introduced in a manner that is consistent with results obtained for the special theory. Thirty-two annulus fibrosus specimens oriented in axial (n = 16) and radial (n = 16) directions were obtained from the middle-lateral portion of intact intervertebral discs from human lumbar spines and tested in a stress-relaxation protocol. Material constants are determined by fitting the theory to experimental data representing the equilibrium stress versus stretch and the surface stress time history curves. No significant differences in material constants due to orientation existed, but significant differences existed due to the choice of theory used to fit the data. In comparison with earlier studies with healthy annular tissue, we report a lower aggregate modulus and a higher initial permeability constant. These differences are explained by the choice of reference configuration for the experimental studies. PMID- 10834160 TI - Unconfined compression of articular cartilage: nonlinear behavior and comparison with a fibril-reinforced biphasic model. AB - Mechanical behavior of articular cartilage was characterized in unconfined compression to delineate regimes of linear and nonlinear behavior, to investigate the ability of a fibril-reinforced biphasic model to describe measurements, and to test the prediction of biphasic and poroelastic models that tissue dimensions alter tissue stiffness through a specific scaling law for time and frequency. Disks of full-thickness adult articular cartilage from bovine humeral heads were subjected to successive applications of small-amplitude ramp compressions cumulating to a 10 percent compression offset where a series of sinusoidal and ramp compression and ramp release displacements were superposed. We found all equilibrium behavior (up to 10 percent axial compression offset) to be linear, while most nonequilibrium behavior was nonlinear, with the exception of small amplitude ramp compressions applied from the same compression offset. Observed nonlinear behavior included compression-offset-dependent stiffening of the transient response to ramp compression, nonlinear maintenance of compressive stress during release from a prescribed offset, and a nonlinear reduction in dynamic stiffness with increasing amplitudes of sinusoidal compression. The fibril-reinforced biphasic model was able to describe stress relaxation response to ramp compression, including the high ratio of peak to equilibrium load. However, compression offset-dependent stiffening appeared to suggest strain dependent parameters involving strain-dependent fibril network stiffness and strain-dependent hydraulic permeability. Finally, testing of disks of different diameters and rescaling of the frequency according to the rule prescribed by current biphasic and poroelastic models (rescaling with respect to the sample's radius squared) reasonably confirmed the validity of that scaling rule. The overall results of this study support several aspects of current theoretical models of articular cartilage mechanical behavior, motivate further experimental characterization, and suggest the inclusion of specific nonlinear behaviors to models. PMID- 10834161 TI - Pulsed laser-induced thermal damage in whole blood. AB - An investigation of the effects of laser irradiation with a wavelength of 532 nm and pulse duration of 10 ms on whole blood was performed in vitro. Threshold radiant exposures for coagulation were quantified and transient radiometric temperatures were measured. The progression of effects with increasing radiant exposure--from evaporation to coagulation-induced light scattering to aggregated coagulum formation to ablation--is described. Results indicate that coagulation and ablation occur at temperatures significantly in excess of those assumed in previous theoretical studies. An Arrhenius rate process analysis based on hemoglobin data indicates good agreement with experimental results. PMID- 10834162 TI - Design and validation of a machine for reproducible precision insertion of femoral hip prostheses for preclinical testing. AB - Preclinical testing of orthopaedic implants is becoming increasingly important to eliminate inferior designs before animal experiments or clinical trials are begun. Preclinical tests can include both laboratory bench tests and computational modeling. One problem with bench tests is that variability in prosthesis insertion can significantly influence the failure rate; this makes comparison of prostheses more difficult. To solve this problem an insertion method is required that is both accurate and reproducible. In this work, a general approach to the insertion of hip prostheses into femoral bones is proposed based on physically replicating an insertion path determined using computer animation. As a first step, the seated prosthesis position is determined from templates and femur radiographs. Three-dimensional images of the prosthesis and bone are then imported into computer animation software and an insertion path in the coronal plane is determined. The insertion path is used to determine the profile of a cam. By attaching the prosthesis to a carriage, which is pneumatically moved along this cam, the required insertion motion of the prosthesis in the coronal plane can be achieved. This paper describes the design and validation of the insertion machine. For the validation study, a nonsymmetric hip prosthesis design (Lubinus SPII, Waldemar Link, Germany) is used. It is shown that the insertion machine has sufficient accuracy and reproducibility for preclinical mechanical testing. PMID- 10834163 TI - [Usefulness and pitfalls in clinical electrophysiological studies]. AB - Clinical electrophysiological studies(EPS) have become useful tools for specific cardiac arrhythmias. However, these studies are expensive and occasionally cause major as well as minor complications such as serious arrhythmias, cardiac tamponade or thromboembolism. It is important to consider guidelines for clinical application of invasive studies. The indications for clinical electrophysiological studies have been categorized into three classes. Class 1: All experts agree that EPS is useful and important for patients treatment and patients with these condition benefit from EPS. Class 2: There is less certainty about usefulness of the information from EPS. Class 3: EPS does not provide useful information. EPS should be performed by trained cardiologist in adequately equipped laboratory. Such studies are performed either by intracardiac pacing and recording or esophageal pacing and recording, and evaluate electrophysiological properties such as automaticity, conduction and refractoriness; initiate and terminate tachycardias; mechanism of tachycardia by activation mapping and pace mapping; evaluate for antiarrhythmic drug, catheter ablation, and intracardiac defibrillator. EPS is useful tool for analysis of bradyarrhythmias and tachyarrhythmias and bring us useful information for the choice of treatment but has its limitation and pitfalls. In general, EPS is not so important to determine the indication of pacemaker for bradyarrhythmias such as sick sinus syndrome or atrioventricular(AV) block. But, it is very useful to analysis tachyarrhythmias and indicate antiarrhythmic drugs or catheter ablation for patients with Wolf Parkinson-White(WPW) syndrome, AV nodal reentrant tachycardia, atrial tachycardia, atrial flutter, or ventricular tachycardia to cure the tachyarrhythmias. PMID- 10834164 TI - [Utility and limitations of ambulatory blood pressure monitoring and tilting test: evaluation of baroreceptor reflex sensitivity]. AB - Ambulatory blood pressure monitoring(ABPM) permits 24-hour measurement of blood pressure(BP) without restricting the subject's activities. The tilting test is used to evaluate BP variability and autonomic nervous responses, especially baroreceptor reflex sensitivity(BRS) during changes of body position. The change of BP and BRS are common to both ABPM and the tilting test. In this instance, BRS is expressed as the ratio between heart rate(HR) variation and systolic BP variation(delta RR/delta SBP). The results were the same as those produced by all other tests including sequential method, squatting test, the neck chamber method, and tilting test. Therefore, we investigated the correlation between BRS and BP changes monitored mainly by means of ABPM. Twenty-four-hour BP variation(SD) manifest both short-term variability(SDh), and long-term variability extending over 24 hours(SD24) (SD2 = SDh2 + SD24(2). In the former case(SDh), atherosclerosis and lowered BRS participate, and in the latter case(SD24), activation of the sympathetic nervous system(SNA) participates strongly. Each BP value during 24 hours(Pi: i = 1, 2, ... 10(5)) is expressed by the product(Pi = phi i x P0) of the ratio of variability(phi i) and sleep-time base BP(P0) values. The phi i is expressed by HR variation and two elements: the cardiovascular response element and the BRS element. In mild cases of essential hypertension, the correlation between Pi and HR variation is high. In severe cases of essential hypertension and in cases of Shy-Drager syndrome, short-term phi i caused by BRS decrease is large, and the correlation between phi i and HR variation during 24 hours drops. Although measuring short-time variability indirectly with ABPM is impossible, BRS can be evaluated on the basis of the product(BI) of diastolic BP(Pd) and pulse interval(RR). The multi-biomedical recorder(TM2425, A&D, Co.) permits 24-hour monitoring of body position and daily activities(acceleration), simultaneous with evaluation of BI, which is related to autonomic nervous activity and BRS. Its employment is, therefore, considered clinically useful. PMID- 10834165 TI - [Head-up tilt test]. AB - Head-upright tilt table testing has been used as a research tool over the past 50 years by physiologists and/or physicians to study the hemodynamic and endocrine adaptation to changes in position. Tilt test has become a widely accepted tool in the clinical evaluation of patients with syncope, since its first application to patients with syncope by Kenny et al. in 1986. There is substantial agreement that tilt table testing is an effective technique for providing direct diagnostic evidence indicating susceptibility to vasovagal syncope. Tilt test duration of 30 to 45 min at 60 to 80 degrees have become widely accepted in laboratories for evaluating adult patients. It exhibits a high level of diagnostic specificity of 80 to 100%. Its sensitivity, in contrast, remains to be 40 to 70%. Pharmacologic provocation during head-up tilt testing elicites susceptibility to vasovagal reactions. Isoproterenol is the most common agent applied for its use. Head upright tilt testing is most warranted in following conditions; recurrent syncope or single episode of syncope in a high risk patient, evaluation of patients in whom demonstration of susceptibility to neurally mediated syncope would affect treatment plans, and exercise induced syncope. PMID- 10834166 TI - [Intravenous contrast echocardiography]. AB - Intravenous contrast echocardiography has become possible in Japan because of the release of the commercially available contrast agent, Levovist. Intravenous administration of Levovist satisfactorily stains the left ventricular cavity, which makes it possible to clearly delineate the endocardial border. Clear delineation of the endocardial border provides easy and accurate measurement of left ventricular dimension and wall thickness, and wall motion abnormalities can be easily and accurately judged, too. Another benefit of intravenous contrast echocardiography is the assessment of myocardial perfusion. Our preliminary experimental and clinical experiences showed the possibility of myocardial staining with intravenous contrast echocardiography. Impressive myocardial staining is obtainable with the combined use of intermittent and contrast harmonic power Doppler imaging. In order to obtain reproducible and clear myocardial contrast images, we have to pay attention to how to inject contrast and settings of ultrasound equipment, i.e., mechanical index, gain setting, depth of focus point, and pulse repetition frequency, artifacts. In the near future, a lot of issues should be standardized to make it possible to compare myocardial contrast echo studies. PMID- 10834167 TI - [Newly developed technology for intravenous contrast echocardiography]. AB - Until now we have not been able to employ a contrast enhancer for ultrasonic echocardiography at the everyday clinical level because the agent itself, composed of microbubbles, was too easily dispersed or even destroyed by several factors. However, contrast echocardiography has made a great leap forward with major developments on two fronts; the application of some new intravenous contrast enhancers, and newly developed machine technology permitting second harmonic imaging, intermittent or triggered imaging, pulse inversion harmonic imaging, and so on. New contrast enhancing agents are proving durability enough to permit greatly enhanced imaging for more than several minutes after injection. Recent new echocontrast specific imaging allows real-time visualizing of myocardial perfusion and assessment of myocardial function. PMID- 10834168 TI - [Usefulness and pitfalls in myocardial perfusion SPECT for detecting and assessing coronary artery disease]. AB - Thallium-201(Tl) is the dominant agent employed for myocardial perfusion imaging for detection of coronary artery disease, assessment of myocardial viability and prognostication. Technetium-99m(Tc) labeled radionuclides have been used as excellent alternatives to Tl. This paper will review the usefulness and pitfall in myocardial perfusion single photon emission computed tomography(SPECT) in patients with coronary artery disease. From a practical standpoint, we should know what are clinical questions, clinical status of patients(history and exercise ability of patients, obesity) and diagnostic accuracy of each diagnostic protocol and the performance in the nuclear laboratory. Myocardial perfusion defects during stress SPECT are produced by a heterogeneity in coronary blood flow, which depends on severity of coronary stenosis and consequent abnormalities in flow reserve. Certain factors can affect sensitivity and specificity of Tl SPECT for detection of coronary artery disease. Accurate determination of myocardial viability is vitally important for clinical decision making for patients with left ventricular(LV) dysfunction who will most benefit from revascularization. Hibernated myocardium may result in profound regional LV dysfunction in absence of necrosis. The various approach such as stress redistribution-reinjection imaging, rest-redistribution imaging and rest redistribution 24 hours delayed imaging has been utilized to assess myocardial viability with Tl. Alternatively, quantitative assessment of 99mTc-methoxy isobutyl isonitrile(MIBI) and tetrofosmin uptake reflect the degree of viability. At the present time one of the most important clinical applications of exercise myocardial perfusion SPECT is the assessment of prognosis for patients with suspected and documented coronary artery disease. Patients with normal stress perfusion SPECT have a low event rate and excellent prognosis. Stress perfusion imagings have been widely used to stratify patients into different risk groups in the United State. PMID- 10834169 TI - [Evaluation of cardiac function by radioisotope angiography]. AB - Cardiac function can be assessed with noninvasive radionuclide technique. Radioisotope angiography can be performed either using equilibrium acquisition or first pass technique. The most commonly generated measurements are as follows; 1) chamber-to-chamber transit time, 2) cardiac output, 3) left-to-right shunt ratio, 4) left and right ventricular ejection fractions, 5) left ventricular end diastolic and end-systolic volumes, 6) regional wall motion, 7) peak ejection rate, and 8) peak filling rate. The ejection fraction, the systolic function and the diastolic function are calculated from ventricular volume curves. The left ventricular volume can be determined by two approaches, the geometric method using the left ventricular image to calculate the area and length of the chamber, and the count-based method. Generally, left ventricular wall motion is observed in right anterior oblique(RAO) view for the first pass method and the left anterior oblique(LAO) view for the equilibrium method. On the other hand, three dimensional multigated imaging was superior in allowing evaluation of left ventricular wall motion in all directions and in assessment of the left ventricular function. Three dimensional multigated imaging is very useful for observation of both left ventricular wall motion and left ventricular function in patients with ischemic heart disease, because it is a noninvasive examination. The quantitative gated myocardial SPECT(QGS) was considered to be useful for assessment of left ventricular function and myocardial perfusion. PMID- 10834170 TI - [Role of intravascular ultrasound imaging for assessing pathophysiology of coronary artery disease]. AB - It has been nine years since intravascular ultrasound imaging(IVUS) was for the first time performed in Japan at our hospital. During this period, the progress of catheter technology brought about many improvements in catheter design and image quality. Also clinical utility of IVUS has been widely recognized with accumulation of clinical experiences. The most important feature of this method is the capability of both quantitative and qualitative analyses of the atheroma. IVUS has mainly been used to help guide procedures during catheter interventions and has provided information about the mechanisms of dilatation and restenosis. Recently, the ability of IVUS in diagnosing morphologic changes such as compensatory enlargement, vessel shrinkage and plaque rupture has much attention. IVUS should enhance our understanding of the pathophysiology of coronary artery disease. PMID- 10834171 TI - [Coronary angioscopy in ischemic heart disease]. AB - The validity of coronary angioscope was discussed in several clinical settings in this article. First of all, although smooth coronary surface was observed by coronary angiography after the intervention of chronic total occlusion, protrusion or thrombus observed by angioscope occupied the arterial lumen and closed the vessel lumen. After implantation of the stent, lumen was widely opened. Serial angioscopic observation revealed that the neointima became thick and non-transparent until 6 months and then became thin and transparent by 3 years after stent implantation. These findings suggested the validity of angioscope in coronary intervention. Second, coronary angioscope revealed the morphologic and pathologic characteristics of culprit lesions in patients with acute coronary syndrome. The thrombus was observed on the yellow plaque in the culprit lesion. The thrombus formed directly over the plaque was mainly white and red thrombus might be formed after the blood flow was disturbed by the white thrombus formed by the platelet aggregation. These findings imply the importance of anti-platelet therapy after acute coronary syndromes. Finally, the angioscope can detect the minimal changes of the plaque formation, progression and rupture, that is, the entire process of the atherosclerosis. The early detection of yellow plaque might be able to intervent mechanically or pharmacologically to prevent its rupture. PMID- 10834172 TI - [Therapeutic strategy and long-term prognosis of acute coronary syndrome]. AB - The relation between initial therapeutic strategy and long-term prognosis was reviewed in patients with acute coronary syndrome. In the Q wave infarction, the effectiveness of reperfusion therapy has been established. The earlier ischemic myocardium is reperfused, the better is its prognosis. The long-term prognosis becomes better when TIMI 3 flow is obtained after the reperfusion therapy. Direct PTCA, which can obtain TIMI 3 flow in more than 90% of the patients, has showed lower mortality, lower rate of reocclusion and lower rate of cerebrovascular accident than thrombolytic therapy. As for the long-term efficacy of stent implantation, several more years are needed although early outcomes of primary stenting are impressive. The lesions in patients with non-Q wave myocardial infarction or unstable angina frequently show severe stenosis with platelet-rich thrombi. This type of lesion is thought to be unstable and the chance of cardiac events are high when percutaneous intervention or bypass surgery is performed during this period. Thus, the conservative strategy with waiting for 1 to 2 weeks to stabilize the plaque has been recommended. However, in the long span, the prognosis of unstable angina seems not bad after the complicated early course. The long-term prognosis of unstable angina could be affected by the early outcome. PMID- 10834173 TI - [Treatment of choice and long-term prognosis for ventricular arrhythmias]. AB - Treatment of choice and long-term prognosis of the patients with ventricular arrhythmias are described in terms of prevention of sudden cardiac death and/or recurrence of life-threatening arrhythmias(ventricular tachycardia and ventricular fibrillation). 1) As to the long-term prognosis of ventricular tachyarrhythmias, presence of organic heart disease and degree of cardiac dysfunction are major determining factors. 2) The prognosis of patients with ventricular arrhythmias depends on how sudden cardiac death and life-threatening arrhythmias can be prevented. Among various methods, the electrophysiological test and its guided-therapy for antiarrhythmic drugs are now believed to be the most effective method for the prediction and prevention of the life-threatening events. We propose that the signal averaged electrocardiography is the best screening method as non-invasive approach for the selection of patients undergoing the electrophysiological test. 3) There are still certain limitations as to the prediction of sudden cardiac death and/or prevention of recurrent life threatening arrhythmias by antiarrhythmic drug treatments in the certain numbers of patients depending on their basal cardiac disease and functional impairment. At present, catheter ablation procedure and implantable cardioverter defibrillator are the choice of the treatment in these cases. 4) In addition to conventional antiarrhythmic drugs, the treatment for the basal cardiac condition is mandatory for the long-term prognosis in the patients with ventricular arrhythmias. PMID- 10834174 TI - [Long-term outcome of pharmacological and nonpharmacological treatment for ventricular arrhythmias]. AB - Recent advances of nonpharmacological therapy such as catheter ablation and implantable cardioverter defibrillator and lessons from the Cardiac Arrhythmia Suppression Trial(CAST) have changed the strategy for ventricular arrhythmias. The safety and efficacy of radiofrequency catheter ablation of symptomatic sustained monomorphic ventricular tachycardia without structural heart disease has made ablation the firstline curative therapy. In idiopathic ventricular fibrillation such as Brugada syndrome, an implantable cardioverter defibrillator is the most effective treatment to prevent sudden cardiac death. In patients with asymptomatic ventricular tachyarrhythmias in heart failure, class I antiarrhythmic drugs should be avoided due to proarrhythmic and negative inotropic effects that may be responsible for increased mortality in some trials. In such patients, amiodarone and beta-blocker may reduce sudden cardiac death. For patients with sustained ventricular tachycardia or ventricular fibrillation in heart failure, amiodarone or implantable cardioverter defibrillator should be considered. In comparison with amiodarone, implantable cardioverter defibrillator markedly reduced sudden death in ventricular tachycardia and ventricular fibrillation survivors in Antiarrhythmics Versus Implantable Defibriltors(AVID). Although better patient selection and clarification of mapping criteria improved the successful ablation rate in patients with structural heart disease, candidates of ablation are few. In patients with extensive structural heart disease, multiple ventricular tachycardias are often present. Catheter ablation of a single ventricular tachycardia may be only palliative. Therefore, implantable cardioverter defibrillator is the most effective treatment to prevent sudden cardiac death, with amiodarone and ablation as the adjunctive therapy to prevent frequent ventricular tachycardia. Furthermore, an implantable cardioverter defibrillator improved survival in selected patients with depressed ventricular function after myocardial infarction, who also have nonsustained and inducible sustained ventricular tachycardia in Multicenter Automatic Defibrillator Implantation Trial(MADIT) and Multicenter Unsustained Tachycardia Trial(MUSTT). PMID- 10834175 TI - [Changes of serum hepatocyte growth factor in coronary artery disease]. AB - Hepatocyte growth factor (HGF) is an endothelial cell specific growth factor involved in the repair of endothelial cells and collateral formation, however, the role for coronary artery disease is still unknown. We measured serum HGF level in various coronary artery diseases to examine the clinical significance. Serum HGF level was measured using the enzyme-linked immunosorbent assay method in patients with stable effort angina pectoris (n = 26), old myocardial infarction (n = 18), unstable angina pectoris (UAP; n = 10) and acute myocardial infarction (AMI; n = 21). As a control group, we selected 11 patients with neurocirculatory asthenia. Blood samples from peripheral veins were collected at cardiac catheterization before heparin administration. In the AMI group, blood samples were also collected at 48, 72 hr, 1, 2, 3 and 4 weeks from the peripheral veins and 48 and 72 hr after reperfusion from the coronary sinus. Serum HGF level was significantly higher in the UAP (0.41 +/- 0.12 ng/ml, p < 0.001) and AMI groups (0.38 +/- 0.26 ng/ml, p < 0.05) compared to the control group (0.19 +/- 0.09 ng/ml). Serum HGF level peaked 48 hr after reperfusion in both the peripheral veins (0.42 +/- 0.16 ng/ml) and coronary sinus (0.58 +/- 0.23 ng/ml) in the AMI group, with a significantly higher level in the coronary sinus than the peripheral veins (p < 0.05). No significant correlation between peak HGF level in the peripheral veins and peak creatine kinase (CK), CK-MB, ejection fraction and cardiac index was observed. Serum HGF was elevated in acute coronary syndrome, indicating advanced endothelial cell damage. HGF is produced, at least partially, in the heart in patients with AMI. Serum HGF level may be useful to detect endothelial cell damage rather than myocardial cell damage. PMID- 10834176 TI - [Clinicopathological study of left ventricular remodeling after first acute myocardial infarction]. AB - The morphological characteristics of post-infarction ventricular remodeling were determined by comparison of infarct location and histological changes of noninfarcted myocardium at autopsy. A total of 94 cases of first acute myocardial infarction with clinical courses of 0 to 37 days were studied. Hearts were sliced on the short axis at the level of 1/3 of the distance from the atrioventricular ring to the apex. Wall thicknesses of the infarcted and noninfarcted areas, and the endocardial and epicardial perimeter lengths of the left ventricle were measured. Myocyte diameter and number of myocytes in the noninfarcted area were measured. Infarcts were classified into 3 groups based on infarct location (51 anterior, 22 posterior, and 21 nontransmural circumferential) and each group was further divided according to the clinical course of less than 72 hours or longer. Fifty two patients died within 72 hours. Cardiac rupture was the most common cause of death in the anterior group. Patients in the posterior group chiefly died due to cardiogenic shock and in the circumferential group chiefly died to pump failure. According to the number of stenosed coronary arteries, cardiac rupture was the most common cause of death in single-vessel disease in both anterior and posterior groups (62.1% and 55.6%, respectively). In double-vessel disease, the most common cause of death in the anterior group was still cardiac rupture (50.0%). On the other hand, 50.0% of the posterior group died of cardiogenic shock in double-vessel disease. Patients with triple-vessel disease mainly died due to heart failure in all groups. Wall thickness of the infarcted myocardium was decreased in the anterior group after 72 hours (11.8 +/- 3.5 vs 7.8 +/- 2.5 mm). Endocardial perimeter length was increased in the anterior and circumferential groups (83.6 +/- 25.6 vs 116.3 +/- 29.5 mm, 75.2 +/- 12.0 vs 117.6 +/- 3.1 mm, respectively). Endocardial/epicardial perimeter length ratio increased with longer clinical course in the anterior group. No specific change in wall thickness or perimeter length was found in the posterior group. Noninfarcted wall thickness was preserved in both the anterior and posterior groups. Myocyte diameter and number of myocytes in the noninfarcted area showed no significant difference after 72 hours. The nature of ventricular remodeling differs with infarct location. Ventricular dilation occurred during the clinical course in the anterior group. The transmural and adjacent areas are more important than the remote noninfarcted area in post-infarction remodeling within this period. PMID- 10834177 TI - Efficacy of primary percutaneous transluminal coronary angioplasty for acute myocardial infarction in patients aged > or = 80 years. AB - The therapeutic result of primary percutaneous transluminal coronary angioplasty (PTCA) in the elderly was assessed in 20 of 44 patients > or = 80 years old (range 81-92 years, mean age 84 +/- 3 years) with acute myocardial infarction who underwent primary PTCA and the results compared with 194 younger patients (< 80 years) who underwent primary PTCA during the same period. In-hospital prognosis, and long-term outcomes were investigated. Emergency coronary angiography was performed in 47.7% of the patients with acute myocardial infarction in the elderly group with 45.5% receiving primary PTCA. Primary PTCA was successful in 95.0% of the elderly group, showing no significant difference from the younger group. In-hospital mortality showed no significant difference between the elderly group and the younger group (5.0% vs 4.1%). However, the incidences of pump failure of the heart and bleeding complication requiring blood transfusion were higher in the elderly group during hospitalization. The 2-year survival rate for the elderly group was 82.5 +/- 9.3%, which was better than previous studies. These results suggest that primary PTCA has a good success rate in the elderly patients with acute myocardial infarction, and decreases the in-hospital mortality and improves the long-term outcome. However, this study was conducted retrospectively in a small group of patients. The efficacy of primary PTCA for the elderly remains to be clarified in a larger prospective trial. PMID- 10834178 TI - Myocardial scintigraphy using iodine-123 15-(p-iodophenyl)-3-R, S methylpentadecanoic acid predicts the response to beta-blocker therapy in patients with dilated cardiomyopathy but does not reflect therapeutic effect. AB - Myocardial fatty acid metabolism is disturbed in patients with idiopathic dilated cardiomyopathy. Myocardial scintigraphy using iodine-123 15-(p-iodophenyl)-3-R, S methylpentadecanoic acid(BMIPP) was used to assess the response to beta-blocker therapy in 19 patients with dilated cardiomyopathy. BMIPP myocardial scintigraphy was performed before and 6 months after initiating beta-blocker therapy with metoprolol. Cardiac BMIPP uptake was assessed as the total defect score (TDS) and heart-to-mediastinum activity (H/M) ratio. Patients were classified retrospectively as responders with an improvement of at least one functional class (New York Heart Association) or an increase in ejection fraction of > or = 0.10 at 6 months, or as nonresponders meeting neither criterion. Responders had a significantly better pretreatment TDS (p < 0.005) and H/M ratio (p < 0.0001) than nonresponders. TDS exhibited no significant changes over 6 months in either group (responders: 13.2 +/- 3.7 vs 12.5 +/- 3.3; nonresponders: 20.8 +/- 6.5 vs 20.5 +/ 3.0). Responders showed no significant changes in H/M ratio (2.47 +/- 0.28 vs 2.43 +/- 0.42); paradoxically, nonresponders showed a significant increase from 1.82 +/- 0.11 to 2.10 +/- 0.19 (p < 0.05), suggesting that beta-blocker therapy protected the myocardial fatty acid metabolism even in the absence of clinical improvement. BMIPP myocardial scintigraphy provides a prediction of response to beta-blocker treatment, but does not reflect the therapeutic effect in responders at 6 months. PMID- 10834179 TI - [Evaluation of cardiac autonomic nerves by iodine-123 metaiodobenzylguanidine scintigraphy and ambulatory electrocardiography in patients after arterial switch operations]. AB - The autonomic cardiac nerves reach the heart after passing through the vicinity of the aortic root and the pulmonary trunk. The arterial switch operation (ASO) completely transects the ascending aorta and the pulmonary trunk. Therefore, this surgical procedure virtually denerves the heart. Cardiac sympathetic denervation and reinnervation were evaluated in patients after ASO using iodine-123 metaiodobenzylguanidine (MIBG) myocardial scintigraphy and parasympathetic denervation and reinnervation using ambulatory electrocardiography [Holter electrocardiogram (ECG)]. MIBG scintigraphy was performed in 14 patients who underwent ASO (ASO group) and 3 patients who underwent other open heart surgery (control group). All patients in the ASO group underwent the operation in the neonatal or infantile period. Planar and single photon emission computed tomography (SPECT) images of the myocardium were obtained. Defect score was determined by the SPECT images as a semi-quantitative index. The mean interval between ASO and MIBG scintigraphy was 25.6 +/- 14.6 months. Holter ECG was also performed in 14 patients in the ASO group and 19 age-matched normal children. The Holter ECGs were plotted on a Lorenz plot. The H index, which is related to vagal tone for the cardiovascular system, was calculated from the R-R intervals. The mean interval between the ASO and Holter ECG was 8.3 +/- 9.7 months. MIBG scintigraphy in the control group demonstrated an almost normal homogeneous tracer uptake, but showed extremely reduced tracer uptake and significantly higher defect score in the ASO group. The extent and degree of the reduction of MIBG uptake improved with time after the ASO. The heart-to-mediastinum MIBG count ratio tended to increase with time. The H index of the ASO group was lower than that of normal children (< 12 months: Control group 0.0280 +/- 0.0068 vs ASO group 0.0219 +/- 0.0083), and gradually increased with time (1-3 years: 0.0470 +/ 0.0157 vs 0.0314 +/- 0.0124). These results indicate that MIBG scintigraphy reflects the presence of sympathetic denervation and the possibility of reinnervation after ASO, and that H index reflects the presence of parasympathetic denervation and the possibility of reinnervation after ASO. These are simple and useful methods for assessing the extent and degree of autonomic denervation and reinnervation. PMID- 10834180 TI - [Methodology of 80 degrees head-up tilt testing with and without low dose isoproterenol provocation in Japanese patients with neurally mediated syncope]. AB - The usefulness of 80 degrees head-up tilt testing with and without low dose isoproterenol provocation was evaluated for the diagnosis of neurally mediated syncope (NMS) in Japanese. Head-up tilt testing was performed in 114 consecutive patients with clinical diagnoses of NMS (68 men, 46 women, mean age 46 +/- 21 years), and 57 times in 36 healthy volunteers (26 men, 10 women, mean age 31 +/- 8 years) who had no history of syncope or presyncope. Head-up tilt testing used an 80 degrees angle for 30 minutes (passive tilt), and if the passive tilt resulted in negative response, isoproterenol was infused at 0.01-0.02 microgram/kg/min and the tilt repeated for 10 minutes (isoproterenol tilt). A positive response was defined as the development of syncope or a presyncopal state associated with hypotension, bradycardia or cardiac arrest. The sensitivities of passive tilt testing for a positive response after 5-, 10-, 15-, 20-, 25- and 30-minute tilting were 1%, 9%, 14%, 19%, 24%, and 28%, respectively, and specificities after 5-, 10-, 15-, 20-, 25- and 30-minute tilting were 100%, 95%, 91%, 88%, 86%, and 84%, respectively. The sensitivities of isoproterenol tilt testing with 0.01 and 0.02 microgram/kg/min were increased to 37% and 48%, respectively. This improvement was statistically significant between the passive tilt and isoproterenol tilt testing with a dose of 0.02 microgram/kg/min (p < 0.01). However, specificities were comparable with those of the passive tilt testing (84% and 82%, respectively). In conclusion, 80 degrees passive tilt testing for 30 minutes showed a low sensitivity (28%) but acceptable specificity (84%). Low-dose isoproterenol provocation was useful for improving sensitivity (48%) while maintaining a comparable specificity (82%). PMID- 10834181 TI - [Risk factors for thromboembolism in patients with atrial fibrillation during treatment with aspirin: a multicenter, cooperative retrospective study. Research Group for Antiarrhythmic Drug Therapy]. AB - Warfarin is effective in preventing thromboembolism in patients with atrial fibrillation, but aspirin is frequently used as an alternative treatment. A multicenter, retrospective study was undertaken to identify patients at risk for thromboembolism during treatment with aspirin. The study group consisted of 470 patients (318 males, 152 females, mean age 59.9 +/- 11.8 years at initial examination) with atrial fibrillation who were treated with aspirin. Thirty-seven percent of patients had paroxysmal atrial fibrillation and 65% of patients received aspirin at a daily dose of 81 mg. Thromboembolism occurred in 31 patients (6.6%) during the follow-up period, resulting in cerebral infarction in 19 patients, transient ischemic attack in 7, and embolism of peripheral arteries in 5. Patients with thromboembolism had lower prevalence of New York Heart Association (NYHA) functional class I (52% vs 72%, p < 0.02) and paroxysmal atrial fibrillation (23% vs 38%, p = 0.085) compared with patients without thromboembolism. Multivariate analysis with the Cox proportional hazard model determined age (> or = 65 years, relative risk 2.29, p = 0.032) as an independent risk factor. NYHA functional class (> or = class II) tended to indicate an increased risk of thromboembolic events (relative risk 1.90, p = 0.076). These results suggest that aspirin has limited efficacy for prevention of thromboembolism in patients with atrial fibrillation who are old (> or = 65 years) or have symptomatic heart failure. PMID- 10834182 TI - [A large right ventricular tumor in a patient with heart failure]. PMID- 10834183 TI - [Left ventricular mass in a patient with leukemia]. PMID- 10834184 TI - Determination of resonant modes of RF heating systems using eigenvalue analysis. AB - In this paper the resonant modes of a radio frequency industrial heating applicator system are determined numerically. This is carried out using a finite element eigenvalue calculation of the electric field of the system. Both the complex linear and nonlinear forms of the generalized large sparse eigenproblem are solved, the latter being obligatory when material properties are frequency dependent. The basis is the Implicity Restarted Arnoldi/Lanczos Methods which can be used to calculate the multiple resonant modes occurring in the operating frequency range of interest. Once the resonant modes of a specific system have been obtained, an electric equivalent circuit model can be derived. The numerical results presented are compared to experimental measurements carried out on a prototype experimental system showing good agreement. PMID- 10834185 TI - A tunable microwave applicator for extraction of date-juice and similar products. AB - A tunable microwave applicator for extraction of date juice and similar products has been developed, designed and constructed. It consists of two main units: a microwave unit and a press/filter unit. The microwave unit consists of a tunable cylindrical cavity resonator, a section of aluminum waveguide with aperture coupling and tuning mechanism and a locally assembled microwave generator working at 2.45 GHz. The press/filter unit is made from a low-loss, heat-resistant material that works inside the cavity. Performance tests at both low and high power levels showed good performance characteristics with power conversion efficiency of more than 70%. Comparison with a commercial microwave oven and an electric heater showed the superiority of the present applicator, in terms of heating rate and power coupling efficiency. PMID- 10834186 TI - Dielectric properties at microwave frequencies studied in partially filled cylindrical TE011 cavities. AB - To measure mean complex dielectric constants of heterogeneous materials with high dielectric losses (like emulsions or granular materials), partially filled TE011 cavities with a time consuming calibration procedure have been used [Persch, 1997], [Risman, 1971], since no suitable theory could be found in literature. In this work, an analytical solution is derived and tested by experiments and numerical results, showing a quantitative agreement, thus making calibration procedures for the measurement of dielectric properties in TE011 resonators unnecessary. PMID- 10834187 TI - Simulation of microwave, conventional and hybrid ovens using a new thermal modeling technique. AB - This paper presents an efficient simulation tool and results for conventional, microwave and combined heating. A new thermal modeling technique for the simulation of conductive and radiant heat transfer is presented. The conductive heat transfer is modeled by a finite difference algorithm. A finite difference scheme is not applicable for the radiant heat transfer, as radiation from a material surface is not bounded to the immediate vicinity as is the conductive transfer. Therefore, ray optical methods are used. Rays connecting mutually visible surfaces are obtained by a new fast method. Some simplifications which are necessary to achieve fast computing are also included. The algorithms are combined with an electromagnetic FDTD program. Simulations are presented for an oven heated conventionally, with microwaves, or by a combination of both. PMID- 10834188 TI - Development of process equipment to separate nonthermal and thermal effects of RF energy on microorganisms. AB - We developed a modified radio frequency (RF) dielectric heater, as a component of a continuous process, for isolating thermal and nonthermal effects of RF energy on microorganisms in liquid foods. The concept combines instantaneous input of RF energy to the food system with rapid removal of thermal energy. We used a double tube heat exchanger as an integral part of the RF heater. The outer tube was Teflon. The inner tube was stainless steel which was grounded in the RF circuit. Product flowed through the annular region between the two concentric tubes. Cooling water flowed through the grounded stainless steel tube. The RF energy was absorbed by the process fluid in the annular region. The cooling water flowing in the inner tube removed the thermal energy from the process fluid controlling the temperature. PMID- 10834189 TI - Synthesis of radial stubs using a numerical model. AB - An optimization approach to the synthesis of radial stubs using numerical model is presented. Microstrip and coplanar waveguide radial stubs are considered. The results are compared in terms of the lengths of the radial stubs. PMID- 10834190 TI - A slotted waveguide field applicator to sustain large diameter uniform plasma cylinders. AB - We describe a slotted waveguide field applicator and present the results of experimental investigation. The experimental results show that it can efficiently transfer power to plasma in the wide pressure range. Specially, it is able to produce a large diameter uniform plasma cylinder with diameter over 160 mm. The electron temperature and density are 2-4 eV and 10(10)-10(11) cm-3, respectively, under conditions of the pressure below 135 Pa and the microwave power 500-900 w. In addition, the primary study of the surface-wave modes indicate that the mode of m = 6 can be excited and propagate. PMID- 10834191 TI - Contribution of whole body F-18-FDG-PET and lymphoscintigraphy to the assessment of regional and distant metastases in cutaneous malignant melanoma. A pilot study. AB - AIM: This pilot study describes use of whole body PET (WB PET) for staging of melanoma. WB PET in conjunction with lymphoscintigraphy (LS) for evaluating status of the sentinel lymph node (SLN) in primary melanoma was investigated with comparison to histopathological results. WB PET was also used both for primary and metastatic melanoma for screening for distant metastases, restaging and follow-up. METHODS: Group I: 17 patients with primary cutaneous melanoma underwent LS, WB PET and SLN dissection. WB PET findings were compared with biopsy results at the SLN site and were used for screening for distant metastases. Group II: 17 patients with a history of melanoma underwent WB PET for follow-up and/or restaging. Results were confirmed or refuted by other radiological modalities or by biopsy or clinical follow-up. RESULTS: Group I: out of 20 SLNs identified by LS in the 17 patients, 18 were negative on WB PET and 2 were positive. 19/20 WB PET findings were confirmed either by histopathology or by clinical follow-up (20 mo). Accuracy was 94% for the assessment of the status of the SLN. Group II: WB PET findings altered staging and treatment in 12/17 patients and confirmed the validity of treatment in 3/17 patients. Overall, in 15/17 patients (88%), WB PET had an impact on treatment strategy. CONCLUSION: Further studies are required to demonstrate if WB PET can become a reliable non invasive alternative to surgery in the characterization of the SLN. WB PET is important as a baseline in primary localized melanoma and decisively impacts patient management in metastatic melanoma. PMID- 10834192 TI - [Diagnostic evaluation of the breast using PET: optimization of data acquisition and postprocessing]. AB - PURPOSE: Development and evaluation of an optimized protocol for PET examinations of the female breast with 2-F-18-fluoro-2-deoxyglucose (F-18-FDG). METHODS: All PET measurements were performed with a whole-body PET system (ECAT EXACT HR+). In order to examine the women with the breasts freely pendant, a special extension for the patient table made of carbon layer composite was designed. After data acquisition in the 3D modus, emission data were sorted into 2D sinograms using the Fourier rebinning algorithm and reconstructed by means of an ultra-fast iterative 2D algorithm (HOSP). The reconstructed emission scans were superimposed onto the corresponding transmission images. The protocol presented was evaluated in examinations on 6 women with breast lesions after the administration of 150 220 MBq F-18-FDG. From two adjacent bed positions, emission and transmission data were acquired over periods of 20 min and 10 min, respectively. For comparison, dynamic magnetic resonance (MR) image series were acquired with a whole-body MR system (MAGNETOM SP 4000) using a double-breast coil. RESULTS AND CONCLUSION: Using the designed extension of the patient table, it was possible to examine corpulent women despite the limited patient port of the PET system in the prone position with the breasts freely pendant. Alongside a reduction in motion artifacts, this positioning also offers the possibility of making a direct comparison between PET and MR images. Despite the fact that the amount of F-18 FDG applied to the patient was markedly reduced, the combination of 3D data acquisition and iterative image reconstruction resulted in excellent quality of the emission scans. By super-positioning of iteratively reconstructed emission and transmission scans, anatomical localization of breast lesions visualized on the emission scans could be improved. The postprocessing of the PET data described was completed in 60 min, this meaning that the presented concept can readily be employed in clinical practice. PMID- 10834193 TI - [Evaluation of a scatter correction technique for single photon transmission measurements in PET by means of Monte Carlo simulations]. AB - PURPOSE: Single photon transmission (SPT) measurements offer a new approach for the determination of attenuation correction factors (ACF) in PET. A major drawback of this method is the high fraction of scattered photons in the transmission sinogram resulting in a marked underestimation of the ACFs. It was, therefore, the aim of the present work, to evaluate a scatter correction algorithm proposed by C. Watson by means of Monte Carlo simulations. METHODS: SPT measurements with a Cs-137 point source were simulated for a whole-body PET scanner (ECAT EXACT HR+) in both the 2D and 3D mode. To examine the scatter fraction (SF) in the transmission data, the detected photons were classified as unscattered or scattered. The simulated data were used to determine (i) the spatial distribution of the SFs, (ii) an ACF sinogram from all detected events (ACFtot) and (iii) from the unscattered events only (ACFunscattered), and (iv) an ACFcor = (ACFtot)1 + kappa sinogram corrected according to the Watson algorithm. In addition, density images were reconstructed in order to quantitatively evaluate linear attenuation coefficients. RESULTS: A high correlation was found between the SF and the ACFtot sinograms. For the cylinder and the EEC phantom, similar correction factors kappa were estimated. The determined values resulted in an accurate scatter correction in both the 2D and 3D mode. CONCLUSIONS: The algorithm proposed by Watson allows an accurate correction of scattered radiation in SPT measurements. The correction factor kappa can by determined experimentally using simple phantoms and then applied to more complex objects. SPT measurements should be performed in the 3D mode, in order to increase the total numb of counts and/or to reduce the measurement time. PMID- 10834194 TI - [Costs of F-18-FDG PET with a satellite concept: update]. AB - AIM: In an earlier study (study I) we had calculated the costs for FDG-PET investigations with our satellite concept (19). 433 patients were included in the study. Compared to the general indications for PET examinations the proportions of investigations were somewhat untypical: Both parameters have changed actually. Therefore we carried out a renewed data analysis (study II). METHODS: All economic parameters were kept, in order to ensure the comparability of both studies. Only an adjustment of the costs to actual conditions was performed. RESULTS: Proportions of investigations: Oncology 63%, Neurology 28%, Cardiology 9%; 3.5 investigations per working day. According to the increased number of investigations the structure of the total costs (DM 1601831) shows a small rise of the percentage costs for the FDG (41% to 42%) and a percentage decrease of the fixed overheads (48% to 43%). The percentage of personnel costs has increased (11% to 15%), the average cost per investigation has diminished: They amounted to DM 2783 in the earlier study (ranging from DM 2458 to 3463). The new data analysis results in the average cost of DM 2366 (-15%) per investigation (ranging from DM 2114 to 2840). Exemplary calculations show that a minimum of the costs will be achieved, if four investigations per day are performed. Retaining all parameters, which do not refer to the costs of the FDG, the average amount for one investigation would vary from DM 2000 (no delivery time) to DM 3300 (4 hours of delivery time). CONCLUSIONS: The results are in good agreement to the general laws of economic science, according to which an increase of the "number of units" with the same fixed overheads will result in a relative reduction of the costs of the "products". A further increase in the frequency of investigations will lead to a rise of the average costs per investigation, because of the physical decay of the FDG delivered once a day. PMID- 10834195 TI - [Investigations of radiation exposure of the medical personnel during F-18-FDG PET studies]. AB - AIM: The aim of the investigation was the identification of those working steps with the highest radiation exposure for the medical personnel during F-18-FDG-PET studies and to evaluate the effectiveness of radiation protection devices and instructions developed in our PET-center. METHODS: The personal dose and hand dose were measured for each working procedure during F-18-FDG-PET studies using electronic personal dosimeters and thermoluminescent dosimeters respectively. Additionally, measurements of the radiation level near the patient were taken. RESULTS: The mean personal dose resulting from syringe preparation was 1 microSv/syringe, from injection 3 microSv/patient, from blood sampling during quantitative studies 6 microSv/study, and from positioning and handling of the patient 6 microSv/study. The mean hand dose per syringe preparation was 710 microSv for each hand. The mean hand dose during injection was 13 microSv for the right hand and 27 microSv for the left hand. All above mentioned values were measured applying the routine radiation shielding in use in our PET center. CONCLUSION: With the developed radiation shielding and means to reduce radiation exposure applied the allowed annual dose for medical personnel are not exceeded. One exception is the hand dose resulting from syringe preparation. An automatic or remote filling device should be used at this working step. PMID- 10834196 TI - [Lymph scintigraphy and pre- and intraoperative gamma probe measurements for localization of sentinel lymph nodes (SLN) in breast cancer]. AB - Aim of this study was to prove the clinical value of nuclear medicine procedures to detect the sentinel lymph node (SLN) for SLN biopsy. METHODS: In 132 patients with breast cancer we performed lymph scintigraphy of the breast as well as both pre- and intraoperative gamma probe measurements correlating the results with the findings of histopathology. RESULTS: SLN were detectable in 62 of 110 patients according to a sensitivity of 56% when scanning was performed only at 1-2 h p.i. while the sensitivity increased to 86% (19 of 22 pts.) if sequential images were acquired up to 2 h p.i. One or more SLN were identified by a hand-held gamma probe transcutaneously prior to surgery in 96% (113 of 118 pts.) of the patients who showed up with no clinically suspected lymph node metastases. Intraoperatively, in additionally 2 patients the SLN could be found resulting in a sensitivity of 97% (115 of 118 pts.). In only 3 patients with clinically no tumor spread to axillary lymph nodes no SLN could be identified by the probe. Skip lesions, i.e. lymph node metastases in patients with tumor-free SLN, occurred in 2 cases: due to SLN biopsy in these patients lymph node staging was false negative compared to conventional staging by means of axillary lymph node dissection. CONCLUSION: The results demonstrate a high preoperative detection rate of SLN in patients with breast cancer using lymph scintigraphy and gamma probe measurements. Thus, nuclear medicine is capable of providing the basic requirements for SLN biopsy in the daily routine. PMID- 10834197 TI - Classification of the lymphatic drainage status of a primary tumour: a proposal. AB - AIM: Creation of a classification of the lymphatic drainage status of a primary tumour. It shall enable comparison of different approaches, standardization and quality control. METHODS: Identification and topographic localization of the sentinel node(s) using lymphatic radionuclide gamma camera imaging and/or gamma probe detection and/or vital dye mapping. RESULTS: A classification comprising four classes (D-Class I-IV) and distinct subclasses (A-E) proved to be simply to be learned and applicable as well as reliably reproducible. It is based on the number of sentinel lymph nodes and their locations and can be combined with the pathological and molecular biological lymph node status. D-classes/subclasses obtained in 420 patients with malignant melanoma of the skin are presented. CONCLUSIONS: The classification is applicable to different approaches. Its diagnostic, therapeutic and prognostic value should be studied prospectively in those primary tumours which preferably metastasis via their draining lymphatic vessels. PMID- 10834198 TI - [Silicosis callosities or bronchial carcinoma: on the problem of functional differentiation using FDG-PET]. PMID- 10834199 TI - [Metabolic imaging of a malignant myxofibrosarcoma with F-18-FDG and C-11 methionine positron emission tomography]. PMID- 10834200 TI - Strength of irradiated bone. AB - A surface of the bone is a site, where the highest mechanical tensions are developed. Therefore, this place has a special importance to provide the strength. The latter depends on a bone tissue atomic/molecular structure. The present report reviews recent results demonstrating how damaged atomic/molecular bonds situated in the surface layer (interface) of the bone have an influence on its strength. The bonds were excited owing to ultraviolet radiation (UR) supplied by a Mercury tube. As the result the strength was decreased. To explore the reason of this phenomenon the atomic force microscopy operated in a tapping mode (AFM) and photoelectron (PE)/exoelectron (EE) analysis was applied. AFM demonstrated a surface topography varied from a homogenize to an island-type under UR. Such a behavior was repeated periodically in monotonically increased radiation exposure. Selective etching of the bone evidenced that reconstruction of the interface layer coupling mineral and organic phases stipulated such a synergetic. EE demonstrated a concentration of destroyed bonds also had a similar regularity depended on exposure. PE measurements discovered responsible for this centres placed at 5.35 and 5.65 eV below a vacuum level. PMID- 10834201 TI - Experiments showing that electromagnetic fields can be used to treat inflammatory diseases. AB - While it is well known that electromagnetic fields (EMFs) can induce repair of non-healing bone fractures, EMF therapy remains confined to orthopedic clinics mainly because the biological and physical mechanisms underlying the therapy are unknown. However, it is generally believed that non-invasive, EMF therapy might have a broad, albeit currently unrecognized clinical potential. In support of this view, we report that 0.1 mT, 60 Hz EMFs induce a 20% mean-increase in anti CD3 binding to T cell receptors (TcRs) of Jurkat cells, a T lymphocyte cell line. Additionally, we show that 60 Hz sinusoidal EMFs and a commercial bone healing EMF modulate signal transduction pathways that regulate lymphocyte proliferation and that are normally triggered by activating the Jurkat TcR. Similar EMF effects are shown in human peripheral blood lymphocytes (hPBLs), exposed to EMFs in culture and in rat PBLs, when donor animals are exposed to a bone healing field (21 days, 4 hr/day). Although we do not yet satisfactorily understand the differences we obtain in cell and animal based experiments, our findings clearly demonstrate that EMFs can regulate lymphocyte proliferation in vitro and in vivo. Since T cells are key modulators of inflammation, the development of EMF based therapeutic devices to regulate their activity can be expected to provide important tools to treat numerous human inflammatory diseases such as psoriasis and arthritis. PMID- 10834202 TI - Morphological and biochemical evaluation of RAW 264.7 cells exposed to polychlorinated biphenyl. AB - The effects of PCBs exposure, at low doses ranging between 10-100 mg PCB (Arclor 1254), on the viability of RAW 264.7 macrophage cells after 24, 48, and 72 hours of incubation was investigated. Furthermore, this study was designed to determine the interrelationship between PCBs and estrogen (E, 10 mg) and its role in the viability of RAW cells. Macrophages were cultured and plated in 24 well-plates according to standard protocols. The wells were divided into eight groups (n = 4 wells/group, 1 x 10(4) cells/well). The cells in groups 1-3 were treated with vehicle (serum-control for estrogen, DMSO-control for PCB, and media alone, respectively). Cells in groups 4 and 5 were treated with low (10 mg) and high (100 mg) doses of PCB. Cells in group 6 were treated with E, and cells in groups 7 and 8 were treated with low dose PCB + E and high dose PCB + E, respectively. Cell viability and damage (Malianodialdehyde, MDA level) were determined after 24, 48, and 72 hours, as well as, cell morphology. The results of this study showed that low and high doses of PCB depressed cell number by 52%. Estrogen treatment caused no effect on cell number in comparison to cells treated with serum alone. Cell number in response to E and low and high doses of PCB decreased cell number by 50%. Similar results were also observed at 48 hour time phase. In contrast, at the 72 hour phase, no significant changes were observed for cell count. Morphological evaluation of the cells revealed healthy spindle shaped multinucleated cells in the control group but, groups exposed to PCB induced morphological changes that included: cells became small, round, and increased evidence of cellular injury and debris. Estrogen treatment did not show changes from the control group. However E and PCB treatments caused the cells to become round, tightly compact nuclei with evidence of cell fragmentation. The results of this investigation showed that exposure to either 10 or 100 mg of PCB had detrimental effects on the RAW 264.7 macrophage cells as early as 24 hours. Combination treatment with E didn't provide any protective measures to the viability of RAW cells exposed to PCBs. PMID- 10834203 TI - The effects of inositol hexaphosphate on the inflammatory response in transformed RAW 264.7 macrophages. AB - Inositol hexaphosphate (IP6) has received much attention for its role in interfering with tumor progression and slowing the metastasis of neoplastic cells. However, there is little information regarding the antioxidant properties of IP6 or its ability to enhance the natural disease resistance of the body. The specific objectives of this experiment were to investigate the effects that IP6 might have on the proliferation and viability of RAW 264.7 transformed macrophages and to morphologically and biochemically investigate the role of IP6 as a free radical scavenger. Transformed RAW macrophages were obtained from the American Type Culture Collection (Rockville, MD) and maintained in sterile media (RPMI) supplemented with 10% fetal bovine serum and 1% antibiotics and antimycotics. The cells were plated on to 24 well plates at a density of 1 x 10(5) cells/well. The cells were divided into five groups of four wells per group per phase (24, 48, and 72 hours). Cells in Group I were treated with media alone and served as controls. Cells in Group II were treated with lipopolysaccharide (LPS) only. Cells in groups III, IV, and V were treated with 1000 microliters of IP6 + LPS, 500 microliters of IP6 + LPS, and 100 microliters of IP6 + LPS, respectively. Cell numbers, as well as, morphology, MDA, and protein were determined at the end of 24, 48, and 72 hours. Data obtained from this investigation revealed that the rate of cell proliferation was totally dependent on the dose of IP6. At 24 and 48 hours and upon the exposure of high dose of IP6 the mitotic ability of the cells was higher (p < 0.05) than the rate at the 72 hour phase. Morphological evaluation of cells at all three phases revealed that there were significant changes in the architecture of cells upon the exposure of IP6 compared to the control group. The results of this study suggest that IP6 may have had an excitatory effect on the inflammatory cell secretions and this phenomenon was found to be dose dependent. PMID- 10834204 TI - Optimal design of connectivity in neural network training. AB - Many authors consider neural network (NN) supervised training as an optimization process during which the weights are iteratively adjusted in order to minimise an error (cost) function, which represents the difference between the obtained and the aimed output. The error function surface is usually nonconvex, can be highly convoluted, with many plateaux and long narrow troughs, and can encounter many saddle and local minima (LM) points. Because backpropagation (BP), which is widely used method for supervised learning, uses local methods for optimization, it can get stuck in LM. This can make learning very difficult and sometimes its convergence to optimal solution--not possible. In this paper we propose a stochastic method for global optimization (GO), which make use of a uniformly distributed LP tau sequence of points. The developed technique is tested with common benchmark problems and used for neural network supervised learning. The conducted tests show that the proposed method can be successfully used for an optimal supervised training of small size NN. PMID- 10834205 TI - Relating induced changes in EEG signals to orientation of visual stimuli using the ESI-256 machine. AB - The focus of this study is to investigate the relations that exist between changes in the orientation of simple visual stimuli displayed to a subject and the induced changes in brain activity recorded as EEG signals. These signals are recorded using the Electric Source Imaging with 256 electrodes (ESI-256). The 256 channel EEG signals of four subjects were measured monopolarly. Each subject was stimulated visually for approximately 7.5 minutes. The stimuli consisted of a series of 300 images depicting four basic orientations of a simple graphical element: a white bar on a black background, with each one of the four orientations (horizontal, vertical, +45 degrees and -45 degrees) shown a total of 75 times in a random order. The notion of missing information under certain orientations is not addressed at this juncture. The EEG signals produced by each subject were recorded in a continuous mode using a sampling rate of 1 kHz. Pre processing of the raw EEG data obtained consisted of epoching, exclusion of faulty electrodes, and reduction of electro-oculogram (EOG) noise due to eye blinks. Topographical maps displaying brain activities and their individual electrode recordings are used as two different means for assessing these changes. It is important to note that the simplicity of the visual stimuli was considered in view of the massive data collected for interpretation. Our goal is to observe and determine new measures that would allow for the quantification and interpretation of such EEG brain activities. Such findings might prove useful for the later use of more complex stimuli and the potential development of size and orientation independent algorithms in image processing. PMID- 10834206 TI - Deriving irradiation control parameters for laser photocoagulation on the retina. AB - Laser photocoagulation is used extensively by ophthalmologists to treat retinal disorders such as diabetic retinopathy and retinal breaks. Currently, the procedure is performed manually and suffers from several drawbacks that a computer-assisted system could help alleviate. Such a system is under development that will rapidly and safely place multiple therapeutic lesions at desired locations on the retina in a matter of seconds. This system provides real-time, motion-stabilized lesion placement for typical clinical irradiation times (100 ms). When performing laser photocoagulation on the retina, the dynamic reflectance of the lesion formed by the laser may be used as a control signal. Using digital filtering and estimation techniques on the sampled reflectance signal, the latency time and growth rate are determined. These parameters are used to derive a control signal for the irradiation time of the laser. The goal is to optimize the therapeutic effect, without overexposing the tissue. The result is consistent lesion formation over any part of the retina. PMID- 10834207 TI - Time-frequency representation of epicardial electrograms during ventricular fibrillation. AB - In the present study we quantified changes in dominant frequency, which is reciprocal of activation interval or cycle period, during ventricular fibrillation (VF). We used a Smoothed Pseudo Wigner Distribution (SPWD) to estimate time-frequency representations of epicardial electrograms recorded in swines. We used a sock with 64 electrodes spaced equally to record electrograms during 30 seconds of electrically induced VF. Results from 29 trials in three animals showed a mean dominant frequency of 6.64 Hz. We observed considerable variation in dominant frequencies during VF. Temporally, the frequencies varied by as much as +/- 1.24 Hz (2 standard deviations). Spatial variation in frequencies was +/- 1.20 Hz. Cycle periods were computed as the reciprocal of dominant frequencies obtained from the SPWD. These cycle periods were verified to be numerically similar to the cycle periods estimated using activation times detected from differentiated electrograms. Results of recent studies by others have shown that cycle periods during VF are correlated with refractory periods. Our results show that a non-stationary analysis technique such as the SPWD can be used to track spatio-temporal variation in cycle periods. These changes can be used to investigate spatio-temporal variation in cellular properties such as the effective refractory periods during VF. The substantial temporal variation in dominant frequencies that we observed suggest the possibility that the excitable gap at any epicardial location also varies considerably from one instance to another during a VF episode. PMID- 10834208 TI - A tree-structured interactive system for representing medical knowledge. AB - In this paper we describe an integration and extension of the software tools in current use by the medical community to provide a sophisticated system for representing and acquiring medical knowledge. For the overall structure of the system that we describe, we propose that a web-based hyperlink system be implemented for representing almost all medical knowledge in a multi-rooted tree structure form. The extension of the database involved to include patient records is discussed. The paper describes the role of contributors to the system as well as users, commentators, and organizers. PMID- 10834209 TI - Use of timelines in senior design--an efficient project management tool for faculty. AB - An important part of any multi-tasked project or undertaking is a work plan. Large contractors with multi-year design projects use a work plan to schedule resources and identify critical design decisions; a smaller company may map out the course and order of work as well as schedule resources. Either way, the work plan is an indispensable tool that assists the project manager in the planning and timely execution of the project. Introduced is the concept of the timeline (or work plan) and its utilization in an Undergraduate Senior Design Course by both Faculty and Student as a Project Management Tool and device from which to chart student progress towards completion of course requirements. Discussed are examples of student project planning and comments with regards to the effectiveness of project planning for the student as well as a discussion of how this device has assisted faculty in charting student progress towards project completion, meeting course requirements, and grading. Project and Time Management issues described in this paper are through the Senior Design Program at the University of Connecticut (UCONN) with National Science Foundation Senior Design Projects to Aid Persons with Disabilities and industry-sponsored projects [1]. PMID- 10834210 TI - Using a multidisciplinary team approach in biomedical engineering senior design. AB - Under Criterion 4, the Professional Component of the Accreditation Board for Engineering and Technology (ABET) Engineering Criteria 2000, a curriculum culminating in a major design experience is required. In addition, under Criterion 3, program graduates must demonstrate an ability to function on multidisciplinary teams. This paper describes some team related issues in senior design at University of Connecticut (UConn) with National Science Foundation (NSF) Senior Design Projects to Aid Persons with Disabilities and industry sponsored projects. While these Criteria can both be satisfied in a senior design course sequence, it is not the most desirable method to achieve satisfactory results. Effective teamwork involves skills that may be learned in classes and training sessions. It is best achieved through team experiences in a series of courses, ideally beginning the freshman year. PMID- 10834211 TI - Developing a controlled vocabulary for use in a hospital information system. AB - In bringing a controlled vocabulary to a health information system, it is important to include those terms commonly used by those who must routinely input data to the system. We have developed a methodology whereby we can obtain "free text" descriptions of diagnoses entered by system users. We then sort those terms/concepts by system and find the appropriate "atomic" term(s). The terms are also being submitted to domain experts for appropriateness and fidelity. These concepts are then coded in an international coding system (SNOMED International) to eventually be entered into the controlled "pick list" of terms available for users to enter. PMID- 10834212 TI - A lumped parameter model of left ventricular filling-pressure waveforms. AB - Assessment of left-ventricular function and quantification of valve stenosis are important in clinical practice as well as in physiological research. This paper describes a mathematical model of diastolic ventricular filling and flow across the mitral valve. The model primarily consists of a system of three first-order, non-linear ordinary differential equations. The equations are solved in MATLAB using the ode45 command. The model generates pressure versus time waveforms for the atrium and ventricle, and flow versus time waveforms for mitral valve flow. Comparisons between model outputs and in-vivo data collected in two porcine experiments show excellent agreement. PMID- 10834213 TI - A nonlinear analytical model of the autonomic basis of heart rate variability (HRV). AB - The sinoatrial (SA) node is responsible for the initiation of cardiac contractions, and integrates charge across its cell membranes on a beat-to-beat basis mainly through the regulation of slow calcium-sodium channels. The time constant (slope) of this integrator and the firing threshold (voltage) of the SA nodal cells are influenced by inputs from both sympathetic and parasympathetic autonomic neural pathways converging on the heart. A model has been developed in Matlab that accounts for cumulative autonomic effects on the SA node's integrator and non-cumulative effects on it's firing threshold, and includes an assessment of the contribution of noise to both sub-systems. Using the model, an assessment of the effect of an arbitrary number of autonomic inputs is made on the beat-to beat variability of heart rate. Additionally, the contribution of noise and nonlinear fast channel effects are considered. Assignment of particular autonomic control frequencies results in physiologically realistic heart rate versus time outputs. Physiological heart rate variability modes such as autonomic control spectra in heart rate, mode-locking, and other complex behaviors can be demonstrated. The results of this model may then be compared with physiological data in which autonomic inputs are controlled by paced breathing or other physiological stimulus. PMID- 10834214 TI - A new device for evaluating autonomic nervous activity of cardiovascular system. AB - The purposes of present study are to evaluate a new measurement device with a piezo sensor to obtain fluctuation of finger blood pressure signal in comparison with the conventional tonometry system. We simultaneously measured continuous blood pressure by tonomery system and finger blood pressure using our device in 12 elderly subjects. Two time series of pulse interval variability (PIV) corresponded to RR interval were estimated respectively as the time between two successive upstrokes of these two devices and systolic blood pressure variability (SPV) was also estimated as the upstroke. In time domain the relative relation of PIV estimated by two systems was high, however, that of SPV was low. On the contrary, in frequency domain, we could estimate autonomic nervous activity of vasomotor activity from our new device. The developed device in our study may be a substitutable device for conventional method as limited to estimate the autonomic nervous activity of cardiovascular system. PMID- 10834215 TI - Evaluating the in situ loading and accelerated durability of barbs located on bifurcated aorto-iliac stent-grafts. AB - A special three-part protocol has been generated for the isolated durability testing of stent barbs. This triple protocol includes an initial evaluation on a cardiovascular duplicator to determine the loading per barb that occurs during normal flow through the stent-graft. The next stage of this protocol determined the frequency response characteristics of the bending point where the barb attaches to the stent to allow for the determination of the appropriate frequency to carry out the accelerated testing. The final part of the test includes high speed bend testing at frequencies determined in the second part of the methods to determine the long term durability of the isolated barb stent unit. The results of this testing indicated that under normal cardiovascular conditions each barb is experiencing a loading of 20 grams peak during maximum forward flow. Loading the isolated barb/stent segment at 600 beats per minute (bpm) for 400 million cycles indicated no tendency for these barbs to experience a change in physical properties. During this testing there were no barbs that broke. PMID- 10834216 TI - Clinical trials of Chitra band material. AB - Stainless steel band material was developed and tested for physico-chemical mechanical toxicological and in-vitro evaluation in dogs, at the Sree Chitra Thirunal Medical Institute, Trivandrum. Weld strength and tensile strength were measured with the Instron testing machine. A clinical pilot study was conducted at College of Dental Surgery, Manipal on 150 patients earlier. In the present study, Chitra bands were cemented on upper and lower molars on one side and Dentaurum bands on the other side, as controls in the same patient. The band was allocated to right and left molars at random. The study was a double blind trial. Thus fifty cases had Chitra bands on the left side and fifty on the tight side. Hundred cases were treated with fixed appliance treatment. The age, sex and technique of treatment were recorded. The spot welder and the cement used were also noted. The patients were recalled every month for nine months and the findings entered in the initial and follow-up proforma by another Orthodontist, making it a blind study. Loose bands, broken bands, broken tubes, broken welds, plaque accumulation, gingival bleeding and gingivitis were noted. The findings will be presented and the clinical applicability of Chitra Band Material discussed. PMID- 10834217 TI - Preparation and characterization of surface-modified calcium hydrogenphosphate by hexamethylene diisocyanates. AB - Isocyanate group has been reported as a coupling agent of hydroxyapatite and polymers. The studies showed that the isocyanate would react with hydroxyl groups of hydroxyapatite and form a covalent bond between isocyanate and hydroxyapatite. In the study, hexamethlene diisocyanate (HMDI) was used as coupling agent. Calcium hydrogen-phosphate (CaHPO4, CHP) powders was the candidate ceramic due to higher content of hydroxyl group, which would react with HMDI at the temperature of 30, 40, 50, 60, and 70 degrees C for 4 hours. Dibutyltin dilaurate and hydroquinone were used as catalyst and inhibitor, respectively. The product was analyzed by DTA, TGA, FTIR, XRD, 13C solid state NMR and 31P, 13C liquid state NMR. From the results, we could prove the surface of calcium hydrogen-phosphate has been successfully modified. The largest amount (5.6 wt%) of HMDI could be grafted on the surface of CHP when reacted at 50 degrees C for 4 hours. Some chain extension could be observed and their structure would also be described in the research. PMID- 10834218 TI - The effects of growth hormone and insulin-like growth factor on the proliferation rate and morphology of RAW 264.7 macrophages. AB - It is well documented that growth hormone (GH) functions to regulate both cell growth and cell number and is considered the master hormone because it affects almost every cell of the body. Growth hormone stimulates the liver to secrete insulin-like growth factor (IGF-1), which is also capable of binding insulin as well as insulin-like binding receptors on the cell surface. It is possible that GH cellular effects are mediated by IGF-1 rather than GH itself. In this study, RAW 264.7 cells were challenged with a high dose of GH (48 ng/microliter), a low of dose GH (4.8 ng/microliter), a high dose of IGF-1 (26 ng/microliter) or low doses of IGF-1 (6.3 ng/microliter) for 24, 48, 72, and 96 hours. Cell number, cell protein concentration, cell damage, and cellular morphology were measured at each time point and compared to untreated RAW 264.7 cells. The results show significant increases in cell number for cells treated with low doses of GH and IGF-1 at 24 hour phase. Cell proliferative effects were also observed at 48 hours in IGF-1 treated cells. Cellular damage (MDA levels) was not statistically significant for any treated group for the entire duration of the experiment. Most notable differences were observed in cellular morphology for both IGF-1 and GH treated cells. IGF-1 resulted in condensation of the nuclear material as early as 24 hours after treatment. IN CONCLUSION: (1) RAW 264.7 responded to both IGF-1 and GH equally (viability and proliferation), and (2) morphological changes were observed in all cells treated with both hormones compared to control group. This study indicates that GH hormone could induce its effect directly or indirectly through IGF-1. PMID- 10834220 TI - Differential effects of ovariectomy on the mechanical properties of cortical and cancellous bone in rat femora and vertebrae. AB - The purpose of this study was to evaluate the effects of ovariectomy on the mechanical properties of bone in the aged Sprague-Dawley rat model of osteoporosis. Eight female rats were sacrificed at the start of the study, at the age of four months. Twenty-four remaining rats were then bilaterally ovariectomized (OVX), and another twenty-four served as controls. Eight rats from each group were sacrificed at five, ten or fifteen weeks. The mean density of L1 vertebral bodies from OVX rats was lower than in their control counterparts, as was the compressive modulus, the ultimate compressive stress and the toughness. The second lumbar vertebra from each rat was cored to remove the bulk of the cancellous bone and tested in compression to failure. Ovariectomy did not affect the mechanical properties of the cored vertebrae, suggesting that the mechanically relevant changes take place in the cancellous bone. The gravimetric density and bone mineral density (measured by dual-energy x-ray absorptiometry) of whole femora were lower in OVX rats than control rats. However, the gravimetric density of the cortical bone was unchanged, and when femora were tested to fracture in three-point bending, the mechanical properties of the midshaft were found to be unaffected. This study suggests some limitations to the mature ovariectomized rat model of osteoporosis: mechanically significant loss of cancellous bone occurs in vertebrae much more rapidly than suggested by other studies. Further, there is little evidence of mechanically important bone loss in cortical bone within four months of ovariectomy. PMID- 10834219 TI - Development of sustained delivery system as a novel technique for tissue culture. AB - One of the greatest advances for biomedical research has been the use of tissue culture to study the effects and the mode of action of various organic compounds. However, there are myriad problems associated with the classical approach to pharmaceutical studies in tissue culture, such as contamination, the logistical problems inherent in intermittent dosing, and near-lethal drug doses required to achieve effective physiological doses. This has given impetus to a search for better means of effective delivery routes in a tissue culture setting. The specific objectives of this investigation were: (1) to develop a ceramic delivery system for the novel purpose of releasing sustained levels of the anti-cancer drug combination inositol hexaphosphate (IP6) + inositol (I) in a tissue culture setting; (2) to assess the proliferation rate and viability of HTB 122 Intraductile Breast Cancer (IBC) cells exposed to sustained levels of IP6 + I compared to conventional means of drug delivery (dissolved in media); (3) to evaluate the morphological changes associated with this treatment process. Sterile aseptic techniques were employed during the entire study (72 hours of incubation). Data obtained from this investigation suggests the following: (1) TCPL delivery system can be utilized in the tissue culture setting to deliver IP6 + I in a sustained manner for 72 hours; (2) the use of IP6 + I at the physiological dose did not induce any significant change in the biochemical marker (MDA); (3) morphological evaluation of cells treated with sustained delivery resulted in cellular atrophy, as well as fragmentation, compared to the treatment by conventional means and to the sham group (empty capsule); and (4) regardless of route of administration, treatment with IP6 + I resulted in a decrease in total protein content compared to the sham capsules and control groups. In conclusion, this approach is a novel use of ceramic delivery systems in tissue culture that gives breakthrough information for basic research on limiting and eliminating contamination and the logistical problems associated with intermittent dosing in tissue culture. PMID- 10834221 TI - Analysis of the shoulder implant. AB - Total shoulder arthroplasty (TSA) is implemented for the relief of pain and better joint function. It is desirable that the TSA can achieve and maintain a long-term and secure fixation of the implanted components. The previous biomechanical studies of shoulder implants were simplified and limited to the two dimensional (2-D) computational finite element analysis (FEA). In this research, reliable 3-D FEA models were established and used to examine the contact stress distribution, contact region and deformation of implant. Based on FEA results, the design of current shoulder implants can be modified for optimum performance. The substantiated 3-D FEA models can provide important engineering data and clinically useful information toward the development of better shoulder implants for a long-term fixation. PMID- 10834222 TI - New UWA robot--possible application to robotic surgery. AB - This research is motivated by the need to design a Nuclear Magnetic Resonance Image guided surgical robot. The achievement of this objective requires the solution of two problems: design and construction of a magnetic resonance compatible mechanical manipulator and development of the appropriate robot control system. It is beneficial to keep robot actuators outside the magnet. Therefore, the parallel architecture should be used for the mechanical manipulator. Newly developed University of Western Australia Robot satisfies this requirement. Moreover, it has substantially larger workspace and torsional stiffness when compared to existing parallel configurations such as the Delta. The plausible method of dealing with the delays in the robot control system caused by the image analysis is the prediction of the deformation based on the mathematical model of the organ mechanical and geometric properties. The hyper viscoelastic constitutive models offer a good way of representing non-linear stress-strain and stress-strain rate relations of soft tissues such as the brain. The numerical values for material constants for brain tissue are given. Additional advantage of the proposed model is that it can be easily implemented in commercially available finite element codes and immediately applied to large scale computer simulations. PMID- 10834223 TI - Morphology of young and old cervical spine intervertebral disc tissues. AB - The intervertebral disc of the cervical spine undergoes degenerative changes during the aging process. Although many studies have reported the qualitative changes in the disc, methodology to quantify these changes is lacking. The present study was designed to quantify the geometrical variations of the nucleus pulposus, annulus fibrosus and uncovertebral joints. The age groups of specimens were classified as juvenile, adult and aged. Fresh intervertebral discs with adjacent vertebral bodies of the lower cervical spine of primates were isolated. The specimens were sectioned sequentially in a coronal plane. Sections were stained with Hematoxylin and Eosin, Verhoeff's, Safranin O, and Trichrome methods to distinguish the nucleus, annulus and uncovertebral joints. Histological images were examined using light microscopy and processed using a computer imaging program to trace the boundaries of the disc components. Dorsal-to-ventral depth and medial-to-lateral width of the nucleus pulposus, and its relative location to the annulus pulposus were also obtained. In the juvenile and adult discs, the nucleus appeared as a light opaque region with scattered notochordal cells with a clear distinction from the annulus region. In contrast, in the aged discs, the nucleus appeared as a dense region of amorphous, irregular collagen materials with less distinction from the annulus region. With the progression of aging, the dorsal-to-ventral depth of the nucleus decreased considerably compared to medial to-lateral width. The uncovertebral joints were clear in the adult discs. The joints were less distinct in the aged discs and their size decreased. Quantification of three-dimensional geometrical variations will assist in better defining the disc tissue in the mathematical models. PMID- 10834224 TI - A novel robust blood pressure control system. AB - A adaptive robust control of postoperative hypertension is presented to reduce the mean arterial blood pressure using vasoactive drugs. It is shown that the control algorithm provides stability-guaranteed regulation subject to infusion rate saturation. Simulation results under clinical conditions show that the control system exhibits strong robustness and is capable of reducing blood pressure. PMID- 10834225 TI - Development of a dye-ligand affinity membrane for the separation of bovine serum albumin. AB - This study reports the development and testing of a dye-ligand membrane. A synthetic dye called Cibacron Blue F3G-A was utilized as the capturing ligand. Bovine serum albumin acted as the target protein. To immobilize the dye molecules, a microfiltration membrane (Versapor) was used as the supporting matrix. The pore size rating of this membrane was reported to be 0.45 micron. Two different methods of dye immobilization were tested. In the first method, the ligand was directly attached to the membrane, whilst in the second method, a spacer molecule was first coupled to the matrix and then dye coupling was carried out. The chemical stability of the adsorbed dye was studied by subjecting the dye membranes to appropriate desorption protocols. As a model affinity system, the adsorption of bovine serum albumin onto the dye-membranes was thoroughly investigated. The equilibrium behavior and the kinetic parameters of the BSA-dye interaction were determined experimentally. PMID- 10834226 TI - The role of the route of administration of poly-chlorinated biphenyls (PCBs) on the reproductive and vital organs of adult female rats. AB - Studies have shown that high doses of Polychlorinated biphenyls (PCBs) given by conventional methods (orally or injections) have adverse effects on the reproductive and vital organs of adult female rats. However, there has not been documentation as to the effects of PCBs on adult female rats by means of a sustained delivery system. The specific objectives of this study were: (1) to investigate the effects of sustained delivery (TCPL ceramics) of PCB versus conventional mode of administration (injection) on the reproductive and vital organs of the adult female rat, (2) to evaluate the role that PCB that might have on the estrus events of adult female rats upon the exposure by sustained delivery (TCPL ceramics) and conventional mode (injection), and (3) to histopathologically evaluate the effect that PCB might have on the ovarian and accessory organs upon the sustained delivery for 21 days. A total of 10 adult female rats (BW 270-300 gm) were randomly divided into three groups. Group 1 (n = 3) served as the control, group 2 (n = 4) was injected intramuscularly every other day with Aroclor 1254 (0.1 cc), and each rat in-group 3 (n = 3) was implanted with TCPL capsules (5 mg of 2,3,3',4,5-Pentachlorobiphenyl). Aseptic surgical techniques were performed throughout the experiment. Blood (1 cc) was collected biweekly for biochemical analysis, and body weights were recorded as well. Pap smears were taken daily at approximately the same time for 25 days, and two slides were made for each pap taken (1 pap stain, 1 Diff Quik). At the end of 21 days post implantation, all control and experimental animals were sacrificed following standard lab procedures (overdose of Halothane). The reproductive and vital organs were collected, weighed, fixed, embedded, sectioned, and stained (H&E) for histological evaluations. Data obtained from this investigation suggest the following: (1) TCPL devices were able to deliver PCB at sustained levels for 21 days, (2) regardless of the route of PCB administration no significant change was observed in total body weight compared to the controls, (3) conventional administration of PCB resulted in a remarkable changes in the fallopian tubes compared to control and sustained delivery implanted animals, (4) there were no obvious change was observed in the phases of estrus cycles upon the exposure of PCB, and (4) histopathological evaluation of spleen, kidneys, heart, adrenals, ovaries, uterus, and cervix tissues exposed to PCB did not reveal any significant changes compared to the intact group. PMID- 10834227 TI - The role of density on the release profiles of hydrophobic and hydrophilic compounds from TCPL delivery systems. AB - Several studies have shown that tri-calcium phosphate lysine drug delivery systems (TCPL) are capable of releasing various organic compounds at a sustained manner for long duration. The fabrication of TCPL devices is complex and the manipulation of various factors can alter drug release profiles. The compression of the TCPL material into the capsular form is an instrumental factor when studying the desired release profile. The objectives of this investigation were: 1) to study the interrelationship between the release profiles of devices that had been cold pressed at two different densities (1.53 +/- 0.15 (Low density, TCPL-LD) and 1.98 +/- 0.7 gm/cm3 (High density, TCPL-HD)), and 2) to evaluate the role of the change in density that might have on the release of hydrophobic and hydrophilic compounds in an in vitro environment. A total of 16 serum bottles were used and later subdivided into four groups (n = 4). Groups I and II contained TCPL-HD and TCPL-LD capsules each loaded with 20 mg of progesterone (P). Groups III & IV contained TCPL-HD and TCPL-LD capsules that were loaded with 20 mg of Bovine Serum Albumin (BSA). Each serum bottle in groups III and IV was filled with 100 mL of phosphate buffer saline (pH 7.4) and 10 microL of sodium azide (antibacterial agent). Serum bottles in groups I and II were filled with 50% alcohol (wt/vol). Samples were withdrawn at various time intervals and the release profiles were analyzed by using standard spectrophotometer techniques. Data analysis was conducted by using Jandel Sigma Stat Statistical software. Results of this investigation suggest that: 1) TCPL-HD and TCPL-LD devices were capable of releasing P and BSA at sustained levels, 2) regardless of the loaded drugs, the release profiles from TCPL-LD devices was found to be significantly higher (p < 0.05) than the release profiles of P and BSA from TCPL-HD, and 3) physiochemical characteristics of the drug to be delivered are instrumental in regulating the rate of release and duration or availability of an effective and safe dose (BSA > P). PMID- 10834229 TI - Optimal autoregressive model based medical image compression using genetic algorithm. AB - Image compression is the process of removal of redundant data in order to achieve savings in storage and communication costs. A new method for predictive coding of images is proposed using two-dimensional multiplicative autoregressive models. Genetic algorithm is implemented to compute autoregressive parameters. Comparison with other multiplicative autoregressive model based image compression method has also been reported here. PMID- 10834228 TI - TCPL drug delivery system: the effects of synthetic DHEA and Diosgenin using an ovariectomized rat model. AB - Dehydroepiandrosterone (DHEA) has been shown in numerous studies to exhibit a host of benefits at the vital and reproductive organ levels. However, the use of naturally occurring DHEA is hindered by its inability to survive the first-pass metabolic process of the liver. One possible alternative means that deserves consideration is the administration of DHEA's precursor, namely, Diosgenin (DG). The specific objectives of this investigation were: 1) to deliver DHEA and DG at sustained levels by Tri-Calcium Phosphate Lysine (TCPL) drug delivery systems using ovariectomized (OVX) adult rats as a model, and 2) to evaluate the biochemical and histopathological changes associated with the sustained delivery of DHEA and DG. A total of 30 adult female rats were used in this investigation. The animals were further divided into 8 groups. Groups 1 and 2 animals served as intact control groups while each rat in groups 3-8 was ovariectomized (sham (33), n = 3 [group 3], sham (47), n = 4 [group 4]). Groups 5 and 6 were implanted with DHEA (group 5) and DG (group 6) loaded TCPL capsules immediately following the OVX procedure. Groups 7 and 8 were implanted with DHEA (group 7) and DG (group 8) loaded capsules 14 days following OVX. Surgical aseptic technique was employed according to standard laboratory protocols. Maliondialdehyde (MDA) and hormonal levels were measured from serum, collected semi-weekly, during the entire investigation (for 47 days) and X-rays were performed weekly. Pap smears were collected daily for 47 days to assess endometrial changes associated with DHEA and DG treatment. Following sacrifice (at day 33 for groups 1, 3, 5, and 6 and at day 47 for groups 2, 4, 7, and 8), routine H and E staining was conducted for histopathological evaluation of the reproductive and vital organs. Results of this investigation demonstrated: 1) OVX resulted in an increase in total body weight, and the use of DHEA and DG returned the body weight to near normal levels as compared to the intact control groups, 2) TCPL capsules delivered DHEA and DG at a sustained level during the 47 day study, 3) serum levels of MDA are as follows: DG > DHEA = OVX > control for the 33 day phase and OVX > DG > DHEA > control for the 47 day phase, 4) no significant changes were observed in total wet weights, as well as the morphology of the spleen, kidney, adrenal, heart, liver, and lung tissues, 5) OVX resulted in an atrophy and non-keratinization trend in the reproductive tissues, and sustained delivery of DHEA and DG showed no remarkable change in these tissues, 6) the use of sustained delivery of DHEA and DG resulted in higher weights of uteri compared to the OVX group. In conclusion, this study provided more information regarding the interrelationship between DHEA and DG, and the physiological responses encountered when they are administered continuously using the adult OVX rat as a model. PMID- 10834230 TI - An eye movement technique for correlating fixational target eye movements with location on the retinal image. AB - Recent investigation demonstrate that ocular motility in eyes with retinal pathology may show a lower propensity to visit such areas of the retina as compared to non-pathological retinal sites. While current ophthalmic instruments with the ability to image both the retina and visual function test target placement on the retina have provided this observation, the ability to quantify in real time these images as an eye movement measurement is presently lacking and the objective of this paper. A Rodenstock confocal scanning ophthalmoscope (CSLO) was used to image the retina during the performance of a visual fixation task. Direct observation of acuity target placement at the retina under continuous viewing conditions was possible with this apparatus. Target fixation eye movement images of the retina were rapidly digitized from video tape records and registered using a specialized rapid retinal image tracking algorithm. Fixational eye movement pattern densities at the retina were derived from these data. Preliminary data obtained demonstrate the utility of this technique in quantifying fixation eye movement patterns observed in three human patients with vocation-related laser retinal injury. In all patients, areas of severe retinal damage was generally avoided. The density of ocular eye movement tends to reflect regions of retinal normality and avoidance of retinal regions with severe pathology. Variation in eye movement density may exist where pathology is less severe. PMID- 10834231 TI - The development of algorithms in electrical impedance computerized tomography. AB - Electrical Impedance Computerized Tomography (EICT) is an imaging method to reconstruct the impedance distribution inside of domain through the boundary injected current and display the impedance contrast ratio as an image. This paper concentrates on developing two algorithms to enhance the quality of the conductivity image. The two algorithms are "Fine-Mesh Conversion Method" and "Sub Domain EICT Method". "Fine-Mesh Conversion Method" is a numerical calibration process to find a coarse mesh impedance network that behaves like a fine mesh network in terms of giving similar voltages under the same current excitations. "Sub-Domain EICT" solves a higher resolution EICT with the cost of a lower resolution EICT by combining "Fine-Mesh Conversion Method", and a Fuzzy Logic Inference Systems (FLIS) classifier. PMID- 10834232 TI - Potential pitfalls of strain rate imaging: angle dependency. AB - Strain Rate Imaging (SRI) is a new echocardiographic technique that allows for the real-time determination of myocardial SR, which may be used for the early and accurate detection of coronary artery disease. We sought to study whether SR is affected by scan line alignment in a computer simulation and an in vivo experiment. Through the computer simulation and the in vivo experiment we generated and validated safe scanning sectors within the ultrasound scan sector and showed that while SRI will be an extremely valuable tool in detecting coronary artery disease there are potential pitfalls for the unwary clinician. Only after accounting for these affects due to angle dependency, can clinicians utilize SRI's potential as a valuable tool in detecting coronary artery disease. PMID- 10834233 TI - Effect of scanline orientation on ventricular flow propagation: assessment using high frame-rate color Doppler echocardiography. AB - Color M-mode echocardiography has recently been utilized to describe diastolic flow propagation velocity (Vp) in the left ventricle. While increasing temporal resolution from 15 to 200 Hz, this M-mode technique requires the user to select a single scanline, potentially limiting quantification of Vp due to the complex three-dimensional inflow pattern. We previously performed computational fluid dynamics simulations to demonstrate the insignificance of the scanline orientation, however geometric complexity was limited. The purpose of this study was to utilize high frame-rate 2D color Doppler images to investigate the importance of scanline selection in patients for the quantification of Vp. 2D color Doppler images were digitally acquired at 50 frames/s in 6 subjects from the apical 4-chamber window (System 5, GE/Vingmed, Milwaukee, WI). Vp was determined for a set of scanlines positioned through 5 locations across the mitral annulus (from the anterior to posterior mitral annulus). An analysis of variance was performed to examine the differences in Vp as a function of scanline position. Vp was not effected by scanline position in sampled locations from the center of the mitral valve towards the posterior annulus. Although not statistically significant, there was a trend to slower propagation velocities on the anterior side of the valve (60.8 +/- 16.7 vs. 54.4 +/- 13.6 cm/s). This study clinically validates our previous numerical experiment showing that Vp is insensitive to small perturbations of the scanline through the mitral valve. However, further investigation is necessary to examine the impact of ventricular geometry in pathologies including dilated cardiomyopathy. PMID- 10834234 TI - Physiological responses associated with sustained delivery of T, DHT, and AED in male rats. AB - The factors responsible for the marked gender difference in the risk of coronary heart disease and remain controversial. Several clinicians and basic scientists support the hypothesis that a protective effect of endogenous estrogen is the key, however epidemiological data failed to prove this trend. The role of endogenous androgens or sex steroid hormones on the development of atherosclerosis has received little attention over the years. The specific objectives of this study are: 1) to deliver testosterone (T), dihydrotestosterone (DHT), or androstenedione (AED) at sustained levels by tricalcium phosphate lysine (TCPL) ceramic delivery devices, and 2) to quantify major biochemical markers levels such as lactate dehydrogenase activity (LDH), total cholesterol, thiobarbituric reactive species associated with sustained delivery of androgens and 3) to evaluate the histopathological changes in vital and reproductive organs associated with excess androgens delivery in adult intact male rats. A total of sixteen male rats were randomly divided into four equal groups. Rats in group I served as intact controls, and animals in groups II-IV were implanted with a single TCPL delivery device contained T (40 mg), DHT (40 mg), or AED (40 mg), respectively. Animals in all groups were weighed and blood was drawn biweekly for cholesterol, LDH and TBAR analysis. At the end of ninety days post implantation, the animals were sacrificed and the organs removed, processed embedded, sectioned and screened for morphological changes. The results of this study suggest that: 1) sustained deliver of androgens caused a statistically significant increase (p < 0.05) in the prostate and the seminal vesicle weights, while the epididymis weights remained similar to control intact animals, 2) decreases in adrenal gland weight was detected in animals treated with androgens compared to control animals, 3) kidney weights increased in all androgen groups compared to control, 4) no differences were observed in heart, spleen, or body weights between the groups, 5) TBAR analysis, as well as LDH activity were not different between androgen treated animals and control animals, and 6) total cholesterol was significantly reduced in T and DHT treated groups compared with control and AED treated animals. The decrease in total cholesterol and reduction in adrenal gland weight may suggest impairment of cholesterol synthesis. In conclusion, this study provides information on the effects of sustained release of DHT, T and AED on the reproductive organs, vital organs, as well as physiological parameters such as TBARS (cellular damage) and total cholesterol. PMID- 10834235 TI - Levels of hydrogen peroxide in tissues adjacent to failing implantable devices may play an active role in cytokine production. AB - Late aseptic loosening of total joint implants continues to be a common cause of implant failure. However, the pathophysiology of implant loosening remains controversial as to which factors at the implant tissue interface plays a crucial role in implant failure. The most prominent features of the foreign body membrane obtained from patients undergoing revision hip surgery were the presence of lymphocytes, histiocytes, giant cells, and immature collagen formation. Biochemical and immunochemical analysis of the tissues revealed increased levels of pro-inflammatory cytokines as well as increased levels of hydrogen peroxide and decreased activity of catalase Increasing concentrations of hydrogen peroxide also caused increases in macrophage release of pro-inflammatory cytokine (IL-1). Macrophage activation by cytokine (TNF alpha) or lipopolysaccharide (LPS) is mediated via translocation of NF kappa B from the cytosol to the nucleus and appears to be dependent upon phospholipase D (PLD). In tissues of patients with aseptic loosening of implants, over production of hydrogen peroxide in response to wear debris stimuli, may activate NF kappa B and initiate cytokine production. PMID- 10834236 TI - Assessment of endometrial function during sustained delivery of estradiol and estradiol plus progesterone in ovariectomized rats. AB - Previous studies conducted in our laboratories have documented that tricalcium phosphate lysine (TCPL) delivery system can be utilized to deliver estrogen (E) and combination of estrogen with progesterone (E + P) at sustained levels for long duration in intact rats. The specific aim of this investigation was to attempt to reverse the endometrial changes resulted from post-ovariectomy by exogenously delivering sustained levels of E and E + P. A total of 27 adult female rats were used in this study. The animals were randomly divided into seven different groups: groups 1 animals served as intact control group, groups 2 and 6 animals were ovariectomized, 3 and 7 were ovariectomized (OVX) and implanted with TCPL loaded with E (20 mg loaded TCPL), and group 4 was OVX and implanted with E (TCPL, 20 mg) + P (TCPL, 40 mg.) treatment. Group 5 animals served, as intact control group. Blood samples were collected biweekly for 47 days. Vaginal smears were taken and screened daily during the entire investigation. Histopathological evaluations were conducted on reproductive as well as vital organs (H & E). Data obtained from this investigation suggest the following: (I) OVX resulted in an increase in total body weight, however E and E + P maintained the body weights at prior ovariectomy level, (II) sustained delivery of E resulted in maturation of vaginal epithelium and the smears exhibited the estrus at the end of 72 hours post implantation and continued this trend for the duration of the study, (III) E + P treatment induced no estrus and the epithelial changes resembled the OVX group, and (IV) E and E + P treatment resulted in a significant different (P < 0.05) in MDA levels compared to OVX and intact control groups. Results of this investigation conclude that sustained delivery of E and E + P by TCPL can be utilized to maintain the normal function of the reproductive organs and could serve as an efficient and safe therapy in estrogen deficient patients. PMID- 10834237 TI - Modeling and optimisation of D-fructose isomerisation using a fluorosensor. AB - D-Fructose was isomerized in a 1.5 litre Tokyo Rikakikai fermentor using Sweetzyme T at 40 degree Celsius at five different feed concentrations ranging from 0.01 to 1.0 M. The stirrer speed was maintained at 100 rpm. The progress of the reaction was monitored using a Dr. Ingold Fluorosensor. The fluorescence voltage was recorded as a function of time and the maximum noted. It was observed that the maximum fluorescence voltage increased as concentration increased from 0.01 to 0.04 and thereafter decreased indicating an optimum of 0.04 M. The fluorescence voltage time data fitted a first order model with an error of less than 0.8 percent. PMID- 10834238 TI - Multi-strategy and multi-algorithm cochlear prostheses. AB - Over the past few years, two generations of cochlear implants have been designed and tested by our group. Since the beginning, our goal was to design a system that is completely transparent to sound processing algorithms and stimulation strategies. The first generation micro-stimulator has eight completely independent channels and the second generation one has sixteen. Each of them is mounted on a thick film hybrid circuit together with necessary interfacing components. The electronic circuits are housed in a hermetically sealed titanium case. They are able to deliver monopolar, bipolar, simultaneous or non simultaneous stimuli on all of their channels. Their electrodes are addressable with all possible combinations. The eight-channel stimulator has already been successfully implanted on volunteer profoundly deaf patients, validating our concepts and our approaches. The second one is endowed with more flexibility and transparency to stimulation algorithms. It offers extended possibilities and more electrodes combinations. In a second version, this one has also been mounted on a thin film hybrid circuit making its size considerably reduced and allowing implanting it in other sites rather than on the temporal bone. The sound processor supporting these systems communicates with internal parts through an inductive link. It is built around a powerful DSP and can be programmed with more than one algorithm at a time. It has been already programmed with all the known strategies and some original ones such as the technique of vector quantization. The latter uses a dictionary of pre-stored stimulation patterns mapped to sound segments. As for programming the systems, this is accomplished by mean of specific user-friendly application software using a window driven completely graphical interface. PMID- 10834239 TI - A curve-fitting technique for evaluating head movement to measure forelimb lameness in horses. AB - Evaluation of the asymmetry of vertical head movement is used during kinematic evaluation of forelimb lameness in horses. For mild lameness, absolute translational measurements are not sensitive enough to detect small differences in the asymmetry of vertical head movement. Also, conscious movement of the head by the horse, not associated with lameness, interferes with accurate measurement. We describe an improved method of evaluating vertical head movement as a measure of lameness, using an model of induced lameness in 9 horses and a time-domain processing technique of curve-fitting. The technique assumes that vertical head movement can be broken down into 3 components; the natural inertially-driven vertical head movement, the alteration of vertical head movement caused by forelimb lameness, and extraneous head movement. The technique uses data from several contiguous strides, eliminates the natural inertially-driven and extraneous head movements, and quantifies lameness as a single value. The technique is shown to more sensitive to change in lameness than absolute measurement of head height translation and to be more accurate than a previously reported frequency-domain technique. PMID- 10834240 TI - Decreased 3D-sound spatialization accuracy caused by speech bandwidth limitation over commodity audio components. AB - This paper studies the accuracy achievable in spatialized speech signals due to their limited bandwidth, in systems based on generic Head-Related Transfer Functions (HRTFs). The accuracy in the perception of sounds spatialized through HRTFs to emulate nine sound locations around the listener, at 0 elevation, are investigated. The tests contrast the localization accuracy for a speech segment and that for broadband noise, as perceived by two independent groups of 10 normal hearing volunteers each. The sound localization perceived and reported by each subject is compared to the HRTF-emulated location, to define a localization error, in each case. The results are analyzed through a repeated measures, mixed factorial design Analysis of Variance (ANOVA). The emulated source locations and the type of sounds are the independent variables, and the average perceived localization error for each case, the dependent variable. Since the spectrum of speech signals drive only sections of the complete HRTF frequency response for each location, the initial expectation is that they benefit less from the spectral-shaping process implemented by the HRTFs, thus resulting in a lower spatialization efficiency than the one for a full-spectrum noise signal. We study the measurement of these effects when 3D-sounds are delivered over commodity audio components, because of their widespread use in practical systems. PMID- 10834241 TI - Autonomic changes during orthostasis: a time-frequency analysis. AB - The transition between sitting and standing represents a period of dynamic changes in cardiac autonomic control. Previous research using heart rate (HR) has suggested a biphasic HR response from sitting to standing with an initial increase in HR followed by an HR decrease before stabilizing at a level above the sitting level. These HR changes have been interpreted as mediated by sympathetic nervous system (SNS) activity. However, more direct measures of cardiac autonomic control have not been investigated in an active orthostasis paradigm. This is due in part to the requirement of stationarity for traditional spectral analytic techniques. In the present study, time-frequency analysis was used to assess the relative contributions of sympathetic and parasympathetic nervous system activity to the cardiac autonomic control during the transition of orthostasis. We examined the heart rate time series during the transition between sitting and standing, and the 60 s prior and the 60 s following the transition. The high frequency component of the HR spectra (an index of parasympathetic activity) decreased sharply at the point of transition and remained depressed relative to the low frequency component (an index of sympathetic activity). These results suggest that time-frequency analysis may be a useful technique to examine non stationary time series. Furthermore, the transition of orthostasis may be mediated more via parasympathetic activity than by sympathetic activity. PMID- 10834242 TI - Numerical simulation of a child restraint system in an aircraft crash-test. AB - Studies conducted at the FAA Civil Aeromedical Institute have shown that when used in aircraft, automotive child restraint devices do not always provide the level of safety desired. Various factors that contribute to poor performance, such as seat belt anchor location, cushion stiffness, and child restraint device design features, were evaluated by a dynamic impact test program. To efficiently continue the research, a computer model was developed using MADYMO. Results of two of the impact tests were used to validate the model. Both test configurations utilized a typical commercial transport airplane passenger seat and a popular automotive child restraint device. These tests were considered representative of the extremes of child restraint device and occupant kinematics due to variance in seat belt anchor location. Details are presented of the test parameters and geometry, as well as cushion and restraint system properties. Test and modelling results for these two impact conditions are summarized and compared. Parametric studies were then conducted that used the model to investigate the effect of cushion stiffness, belt anchor spacing, and initial belt tension. PMID- 10834243 TI - An improved Morris water maze tracking algorithm for psychophysical studies. AB - We have developed an algorithm to track a rat swimming in a Morris Water Maze. It provides an update on rodent position 15 times per second. The tracking algorithm uses a rotation insensitive tracking template coupled with a limited exhaustive search technique to maintain lock on the moving rat. At the completion of the run, the relative time the rat spent in each quadrant is reported. The tracking algorithm was successfully used to evaluate the time course of memory loss and correlate that to tissue loss in the rat cortex following blunt trauma to the sensory cortex. Results of that study are reported in another paper. Our project budget did not allow for commercially available software to perform the tracking task which typically provide position updates 10 times per second. This paper focuses on the tracking algorithm and support equipment used to measure the rats' progress in the maze. PMID- 10834244 TI - Impact simulation on pre-formed prosthesis for large cranial defect. AB - Cranioplasty greater than 10 cm2 in size is challenging task for neurosurgeon. There are many successful trials to fill the small defect with various materials. Proper cranioplasty can provide subsequently restoration of cerebral protection, cosmetic aspect and neuronal function. However, larger defect may not provide adequate protection from the auto, industrial or sport accident even with cranioplasty. In this study, patch design using two popular materials was evaluated and compared with bone material. Then impact-releasing holes were implemented to the patch. The movement of the large patch and its effect on the underlying brain tissue upon simulated impact was evaluated by using finite element analysis. PMID- 10834245 TI - Effects of hyperbaric oxygen on focal brain contusions. AB - The effect of hyperbaric oxygen (HBO) on focal brain contusion was examined for possible prevention of neural degeneration in the central nervous system following blunt trauma. All animals were tested for memory and balance/motor skills, prior to and after injury, using the Morris water maze and beam walk test. Experimental and control groups, each containing 12 juvenile male hooded Wistar rats (Rattus norviegicus), were contused using a 20 g weight dropped from 20 cm onto the dura mater over the somatosensory cortex. The experimental group underwent 45 minute HBO dives at 2.5 atmospheres absolute (ATA) four times a day for 10 days. Memory and motor skills were tested on both groups at 3, 6 and 9 days after injury. Animals were sacrificed and their brains were sectioned and analyzed histologically for neural damage. Examination of the damage to the surface of the brain showed hole sizes in the experimental group were smaller than those in the control group (p = 0.0475). Initial examination of behavioral testing showed mixed results. Beam walk scores 3 days post contusion were significantly better for experimental vs control groups (p = 0.0442) suggesting initial benefits of HBO treatment. However, preliminary analysis of cells in the CA3 region of the hippocampus showed no significant difference between pyramidal cell counts 10 days post-contusion in the ipsilateral sides of either group. PMID- 10834246 TI - A design of an associative memory array processor for ultrasonograph image acquisition and processing. AB - This paper describes a design of an associative memory array processor that can be used in the acquisition and processing of ultrasonograph images. The major concept is to design a parallel architecture that reduces task's execution time by analyzing multiple parts of the image concurrently. The architecture constitutes a distinctive type of single-instruction stream, multiple-data stream machine that is built around content-addressable associative memory slabs, that allow parallel access of multiple memory words. The basic building block of this architecture is a one-pixel processing element, which can perform the standard load (data acquisition) function and also contains some special comparison logic to enable its content to be compared with an external data. Several image processing operations are implemented in parallel, among them: component labeling, size filtering, pattern centralization, and pattern recognition. The proposed novel architecture can label specific regions into the image and isolate them intelligently. It is also capable of storing templates that may be considered as references for similar cases. The system is able to perform learning process and extract features from several input patterns and store the reference pattern in a slice. Moreover, the system is capable of comparing an input image with a pre-stored template during recognition process. The proposed architecture is of interest because it speeds up the recognition process and helps radiology specialists to write their reports confidently. PMID- 10834247 TI - Human nervous system function emulator. AB - This paper describes a modular, extensible, open-systems design for a multiprocessor network which emulates the major functions of the human nervous system. Interchangeable hardware/software components, a socketed software bus with plug-and-play capability and self diagnostics are included. The computer hardware is based on IEEE P996.1 bus cards. Its operating system utilizes IEEE 1275 standard software. Object oriented design techniques and programming are featured. A machine-independent high level script-based command language was created for this project. Neural anatomical structures which were emulated include the cortex, brainstem, cerebellum, spinal cord, autonomic and peripheral nervous systems. Motor, sensory, autoregulatory, and higher cognitive artificial intelligence, behavioral and emotional functions are provided. The author discusses how he has interfaced this emulator to machine vision, speech recognition/speech synthesis, an artificial neural network and a dexterous hand to form an android robotic platform. PMID- 10834248 TI - A system for the ambulatory recording and analysis of nonlinear heart rate dynamics. AB - The study of cardiac dynamics is a topic of great clinic and experimental concern. However, the ability to apply methods from the emerging field of nonlinear dynamics to cardiac time series has been limited by the demand for large amounts of relatively artifact-free data. Both the ability to collect such data and the capacity to analysis such large data sets represents limits. In the present paper we describe a system that allows for the collection of large amounts of high quality data and the analysis of large data sets. The recording system consists of a miniature, single-module electrocardiogram-recording device. This module consists of an integrated three-electrode device that is attached to the chest of the subject. A low power 8-bit micro-controller detects the R-spike and stores the time between R-spikes in milliseconds on a 512 KB EEPROM. This system can record continuously for over four days. The output of this system is down-loaded to a PC and the RR intervals fed to a suite of digital signal processing programs. Among other things, estimates of Approximate Entropy and Poincaire plots are generated. This system will expand the capability of researchers and clinicians to investigate nonlinear cardiac dynamics in ambulatory subjects. PMID- 10834249 TI - Autonomic control of the heart during circular breathing. AB - The analysis of heart rate variability (HRV) has become an increasingly popular method for probing the autonomic nervous system (ANS). A number of specific periodicities have been identified in the spectra of cardiac time series. One major mechanism involved in a number of these periodicities is respiratory modulation of neural traffic to the heart. One respiratory maneuver that has never been investigated is circular breathing. This technique is employed by wind musicians to produce an uninterrupted stream of sound without pauses for breathing. The current study examined the effects of circular breathing on ANS control of the heart in a highly proficient professional musician. Results indicated that circular breathing both with and without producing a sound was associated with altered autonomic control of the heart. Specifically, circular breathing was associated with increases in heart rate and decreases in high frequency HRV. This high frequency HRV has been most closely linked with respiratory sinus arrhythmia and respiratory modulation of vagal neural traffic. We conclude that the examination of novel breathing maneuvers may help to illuminate the role of respiratory modulation of HRV. PMID- 10834250 TI - Algorithms for the computer modelling of mechanical interactions with soft tissue. AB - The computer modelling of soft tissue has applications in the simulation of surgical procedures, product design, and in CAD generally as well as in motion pictures. We begin by reviewing computer modelling techniques in a range of applications. Algorithms are described to predict where surfaces interact mechanically, aiming as far as possible to use geometric modelling methods available within high level CAD systems. After an initial 2-Dimensional evaluation, 3-dimensional surfaces were used throughout. A model was developed to predict the region of impact of two surfaces in an environment of many surface patches. We report the results of this modelling and the application of refinement algorithms to reduce the computation requirements. We then go on to discuss the applicability of this work in the modelling of the deformation behaviour of soft tissues. PMID- 10834251 TI - Material property determination of sub-surface objects in a viscoelastic environment. AB - Modeling human organs and soft tissue or anatomic regions for the purpose of medical training and simulation is a relatively new area. The data presented here is the groundwork for our ongoing development of a real-time haptic virtual environment for abdominal soft tissue palpation. The purpose of modeling the human abdomen is twofold. First, to provide a mathematical description of soft tissue and organs and second, to simulate the behavior of realistic interactions in real-time within a virtual environment. We have developed a, non-invasive, system that will allow us to determine mathematical functions that model the deformation of individual layers of soft tissue within the human abdomen. This system has been tested, experimentally, with a viscoelastic polyester foam model. We have been able to determine the stiffness and force/displacement function of an object beneath two layers of foam having different material characteristics. These experimental results correlate well with known polyester foam material characteristics. In general, the calculated stiffness constants were within 5% of the actual value. The data presented in this study shows that this system may be a viable tool for accurate measurement of human soft tissue properties and behavioral response to palpation. PMID- 10834252 TI - The application of computer aided product development techniques in medical modelling topic: rehabilitation and prostheses. AB - When considering reconstructive surgery it is often difficult to ascertain the exact nature of affected internal anatomy. Although advances in Computed Tomography (CT) and Magnetic Resonance Imaging (MRI) have enabled the generation of 3D reconstructions of internal anatomy, they are often only available as fixed 2D images. These may obscure important details or prove ambiguous depending upon the angle of view. Another problem is maintaining accuracy of reconstruction that may involve osteotomy, tissue grafts or implants. In such circumstances the ability to plan and rehearse complex procedures can prove invaluable, greatly improving surgical results whilst helping to eliminate potential errors and reducing theatre time. In this context 3D scan data can be treated as the equivalent of Computer Aided Design (CAD) data and can be applied to the production of accurate physical models. The approach undertaken by the DERC is based on close collaboration with surgical departments and enables the rapid and economic production of medical models based on CT/MRI data. This paper illustrates the transfer of product design technologies into a clinical context. Specifically, the use of 3D surface scanning and rapid prototyping technologies as an aid to post operative reconstruction is described. An evaluation of the application of product development tools within a clinical context is presented. The paper concludes with an assessment of likely future application in this area. PMID- 10834253 TI - Modeling a two-mile run age and body weight handicap for men and women. AB - PURPOSE: To develop a weight and age handicap model for Two-Mile Run Test (2MRT) performance for adult men and women using dimensional analysis and recent empirical data. METHODS: Models of the body composition- and physical activity adjusted effect of age on VO2max and the allometric model of body size and 5 K run time for men and women (adjusted for the 2MRT distance) were employed. RESULTS: Two tables were developed, one for each gender. Each provides a simple index (sec) of run time based on body weight and age, that are subtracted from actual 2MRT time to yield adjusted run time. CONCLUSION: Such a handicap model, which grants a handicap based only on the independent effects of age and weight (not fatness or lack of physical conditioning, for example), is the first to be based on physiologically sound theory and empirical data. It may prove useful for military fitness testing particularly in situations where higher achievement is rewarded and/or control for age and weight differences in warranted. PMID- 10834254 TI - Head-neck finite element model for motor vehicle inertial impact: material sensitivity analysis. AB - The aim of this study was to conduct a material sensitivity analysis using a head neck finite element model (FEM). The model included the skull, C1-T1 vertebrae, intervertebral discs, facet joints, and biomechanically relevant ligaments. Poisson's ratio and elastic modulus of the head-neck components were varied. The loading condition included the impact load applied to the first thoracic vertebra. Commercially available software (LS-DYNA) was used for the analysis. Head angle versus time, head center of gravity trajectory, and head center of gravity angular acceleration responses were computed. In general, the variation of elastic modulus had a higher effect on the response compared to variation of Poisson's ratio. As the elastic modulus was increased, the head angle and angular acceleration increased. The present findings form a first step in the study of computational biomechanics of vehicular-related trauma. PMID- 10834255 TI - Finite element model of human cervical spinal column. AB - The purpose of this study was to develop a detailed anatomically accurate finite element model (FEM) of the whole cervical column (C2-T1) using sagittal and coronal computed tomography (CT) scan images and cryomicrotome anatomical sections. The bony vertebrae were defined using CT scan images. The geometrical details of intervertebral discs, uncovertebral joints and ligaments were obtained from cryomicrotome sections. The following steps were used to develop the FEM: wire mesh, surface model and solid model. The wire mesh of each vertebra and disc was generated from the CT and cryomicrotome sections, respectively. The surface model was developed by connecting the wire mesh lines. The solid model was obtained by filling the volume enclosed by the surface model. Finally, the FEM was constructed by discretizing the solid model using the mapped-mesh option. Appropriate finite element types were assigned to each component. For example, the isoparametric eight-noded solid elements were used to define the cancellous bone of a vertebra. The material properties reported in literature were adopted in the model. Commercially available software (IDEAS and ABAQUS) was used to develop the FEM. PMID- 10834256 TI - Long-term sustained delivery of 3'-azido-2',3'-dideoxythymidine in vivo by means of HA and TCP delivery devices. AB - This study will attempt to determine the amount of circulating drug, when provided at sustained levels over an extended period of time, which provides efficacy and reduces the toxic side effects seen with the use of AZT in vivo. Two ceramic delivery systems were selected for this investigation namely tricalcium phosphate (TCP) and hydroxyapatite (HA) ceramic implants. Three different dosages were selected (60, 90, 180 mg AZT). A total of 105 rats were used in this study and they were further divided into seven equal groups (n = 15 per group, 250-300 gm BW). Rats in groups I, II and III were implanted with TCP ceramic capsules containing 30, 60, and 90 mg AZT powder, respectively. Each rat in groups IV, V, and VI were implanted with HA capsules containing 30, 60, and 90 mg AZT powder, respectively. Rats in group VII were unimplanted and served as controls. The standard aseptic surgical techniques were performed in this investigation. Rats were anesthetized with a mixture of Zylazine/Ketamine, and their hind limbs shaved and scrubbed with providone iodine. The sterilized ceramics (gas/24 hours) were inserted under the skin using standard surgical techniques. After implantation, the site was sealed with wound clips, and the animals were injected with 0.1 ml of 200,000 units of Penicillin. At the end of 2, 4, and 6 weeks after implantation, five rats from each group were euthanized and the ceramic capsules, reproductive, and vital organs were removed and examined by following standard laboratory protocols. Results obtained from this study revealed that the rates of AZT released from TCP ceramic implants (30 mg = 2.38 +/- 0.23 ng/ml, 60 = 4.64 +/ 1.03 ng/ml, and 90 mg = 11.92 +/- 2.36 ng/ml serum AZT) were significantly higher than the rates of AZT released from HA ceramic implants (30 mg = 0.84 +/- 0.05 ng/ml, 60 = 2.40 +/- 0.83 ng/ml, and 90 mg = 6.41 +/- 1.24 ng/ml serum AZT). Data obtained from this investigation suggest that: (I) TCP and HA ceramic implants can be used effectively to deliver AZT in amounts capable of eliciting physiological responses in vivo, and (II) large fluctuations of AZT concentrations in the blood stream and tissues due to the conventional routes of administration could be eliminated by using ceramic sustained delivery system. PMID- 10834257 TI - Cytomorphological assessment of benign and malignant dense hyperchromatic groups in cervicovaginal smears. AB - Dense hyperchromatic cell groups are considered common diagnostic problems in cytopathological evaluations. Clusters of cells with scant cytoplasm and dark nuclei represent the morphological features of dense hyperchromatic cells. Cytological evaluations of the dense hyperchromatic groups in cervicovaginal smear results in high rates of false-positive or false negative diagnosis. The key element is to differentiate among the dense hyperchromatic groups and to appropriately classify, based on strict morphologic criteria. The objective of this study was to evaluate the cytomorphology of benign and malignant dense hyperchromatic groups in cervicovaginal smears reported at the University of Mississippi Medical Center. Six distinct types of dense hyperchromatic groups were selected (forty-eight cervicovaginal smears) to represent all of the entities. The cases were divided into; group 1; atrophic pattern (n = 9), group 2; endocervical cells (n = 9); group 3, endometrial cells (n = 10), group 4; high grade squamous lesions (HSIL)(n = 10), group 5; squamous cell carcinoma (n = 5), and group 6; endometrial adenocarcinoma (n = 5). Light microscope techniques were used to evaluate several parameters--such as, background, arrangement, and chromatin pattern. ImagePro digital analysis computer software (at x40 magnification) was used to quantify and evaluate the nuclear area and nuclear to cytoplasm ratio. Data obtained from this investigation suggest that there were significant differences observed in the total nuclear areas among all groups. In conclusion, cytomorphometric analysis can be utilized as an ideal diagnostic tool in differentiating between the ambiguous or suspicious groups of dense hyperchromatic cells. Ultimately, this diagnostic tool can minimize the rate of false-positive or false-negative diagnosis resulting in better cytologic evaluations and patient management. PMID- 10834258 TI - Morphological and biochemical evaluation of RAW 264.7 macrophages after acute and chronic administration of DHEA and AED. AB - The effect of adrenal hormones namely dehydroepiandrosterone (DHEA, 10-50 ug/day) and androstenedione (AED, 10-50 ug/day), on RAW 264.7 macrophage survival at 24, 48 and 72 hours after lipopolysaccharide (LPS, 2 micrograms/ml) exposure was investigated in an in vitro environment. RAW cells were obtained from American Type Culture Collection and standard laboratory protocols were followed in cells plating (10(6) cells/well), phase terminating, morphological evaluation, and biochemical marker analysis. From physiologic to supraphysiologic doses of DHEA and AED at 24 hours caused increased levels of cellular proteins and cell number without causing any significant (p < 0.05) change in cellular membrane integrity (Maliondialdehyde, MDA) or viability (morphology). At 48 and 72 hours, cells treated with either AED or DHEA did not sustain the increased cellular proliferation as observed at 24 hours and did not significantly differ (p < 0.05) in cellular protein content. RAW 264.7 cells treated with LPS for 30 minutes prior to AED or DHEA exposure caused slight decrease in cell number and cell protein content. The decrease in both cell number and cell protein content were not attributed to increased cell damage or decreases in cell viability due to the fact that the cellular MDA levels were not statistically higher than the control values (p < 0.05). Morphological Evaluations of cells using Image Pro Software, revealed no significant or adverse changes when compared with cells treated with media alone. Dot blot analysis of pro-inflammatory cytokine (TNF alpha) production after LPS treatment was suppressed by DHEA while AED had a minor influence on the responses. These data imply that LPS mediated activation of RAW 264.7 cells can be inhibited by the addition of pharmacological doses of adrenal hormones such as DHEA. PMID- 10834259 TI - Morphological investigation of the rat kidney after continuous administration of testosterone, dihydrotestosterone, or dehydroepiandrosterone using TCPL delivery devices. AB - The response of the kidney tissues to steroids, mainly dehydroepiandrosterone (DHEA), testosterone (T) and dihydrotestosterone (DHT) has been used extensively to investigate the mechanisms of steroid action on hypertension. The specific objective of this study was to investigate anatomical characteristics of kidney tissue after sustained delivery of DHEA, DHT or T at physiological or supraphysiological doses using adult male rats as a model. A total of eighteen adult male Sprague Dawley rats were divided randomly into six equal groups. Animals in groups I-V were implanted intraperitoneally with a single tricalcium phosphate lysine delivery system (TCPL) loaded with T (physiological dose), T (supraphysiological dose), DHT (physiological dose), DHT (supraphysiological dose), and DHEA (physiological dose), respectively. Animals in Group VI were implanted with empty TCPL capsules and served as sham controls. At the end of 21 days post implantation, the animals in each group were sacrificed and the kidneys were harvested, processed, embedded, sectioned and screened for cellular alterations. Data obtained from this study have shown physiological doses DHEA or supraphysiological doses of DHT and T had a significant effect on the areas of the glomeruli, without disruption or changes in area of proximal tubules. Histological evaluation of the organs (5 microns, H&E) revealed nephrotic hypotrophy in all kidneys obtained from animals given supraphysiological doses of DHT or T in comparison to sham animals. PMID- 10834260 TI - Cytohistologic correlation in patients with clinical symptoms of postmenopausal bleeding. AB - Today, the life expectancy for women is longer; therefore, many will likely experience the postmenopausal period (termination of fertility and menstrual bleeding). Uterine bleeding after this period is a sign of pathologic condition. The specific objective of this project was to evaluate the cytohistologic findings in women with postmenopausal bleeding (PMB) and to determine the presence of any significant pathologic lesions. Cytohistologic correlations from 66 patients attained in 1993 from the University of Mississippi Medical Center were evaluated. The population evaluated were divided into three groups: (control group 1) dysfunctional uterine bleeding (DUB), (control group 2) postmenopausal (PMP), and (test group 3) the group of women with postmenopausal bleeding. The DUB and PMP age-matched controls (n = 12, mean age 51 +/- 5 and 57 +/- 5 years) were randomly selected, and correlated with the actual group being tested (54 PMB, mean age 57 years). The distribution among the 54 PMB women evaluated were 69% (37/54) black, and 31% (17/54) white. The DUB and PMP control groups consisted of 50% (6/12) black and 50% (6/12) white, respectively. Histopathological confirmation (62/66--94%) revealed 47/66 as negative, 5/66 as endometrial hyperplasia and 10/66 as squamous cell carcinoma or adenocarcinoma. A significant lesion with endometrial pathology was found in 23% of the patients. These findings suggest that the majority of women in this study with clinical symptoms of postmenopausal bleeding were negative for malignancies. While these results lean more towards a normal cytologic evaluation, postmenopausal bleeding should not be taken lightly. Postmenopausal bleeding could represent signs of more serious lesion such as squamous cell carcinoma or endometrial adenocarcinoma if not detected and managed early. PMID- 10834261 TI - A microcomputer-based data acquisition system for ECG, body and ambient temperatures measurement during bathing. AB - A data acquisition system employing a low power 8 bit microcomputer has been developed for heart rate variability monitoring before, during and after bathing. The system consists of three integral chest electrodes, two temperature sensors, an instrumentation amplifier, a low power 8-bit single chip microcomputer (SMC) and a 4 MB compact flash memory (CFM). The ECG from the electrodes is converted to an 8-bit digital format at a 1 ms rate by an A/D converter in the SMC. Both signals from the body and ambient temperature sensors are converted to an 8-bit digital format every 1 second. These data are stored by the CFM. The system is powered by a rechargeable 3.6 V lithium battery. The 4 x 11 x 1 cm system is encapsulated in epoxy and silicone, yielding a total volume of 44 cc. The weight is 100 g. PMID- 10834262 TI - Time averaged spatial distribution of epicardial dominant frequencies during ventricular fibrillation. AB - Recent evidence suggests that the dominant frequencies during ventricular fibrillation (VF) may be used as indicators of dispersion in repolarization and in activation patterns. In the present study, we quantified dominant frequencies from multiple epicardial electrodes to investigate if there are differences in the averaged frequencies within the electrograms recorded from the left and the right ventricles. Further, we quantified whether the difference in average frequency between the two ventricles changed during 30 seconds of VF. Results from eighteen trials in two pigs showed that during the entire duration of VF the average dominant frequencies of all electrodes over the left ventricle were higher than those over the right ventricle (p < 0.005). The dominant frequencies are reciprocal of cycle periods or activation intervals during VF. Our results show that on average, activations in the left ventricle occurred at a faster rate than those in the right ventricle. Activation intervals at any site are determined by the refractory period at that site and the arrival time of next activation. Although differences in cellular properties may have contributed to the observed differences in activation intervals between the ventricles, it is possible that activation arrival times may be different as well. It is possible that the increased tissue mass of the left ventricle may increase the probability that any site will get excited at a faster rate after it is recovered from previous activation. PMID- 10834263 TI - Effects of high fat or high sucrose diets on rat femora mechanical and compositional properties. AB - Diets high in fat and/or sucrose decrease whole bone mechanical properties and mineralization. This study examines the impact on rat bone mechanical properties and composition of age and diets (a) low in fat, (b) high in sucrose, and (c) high in fat. Sprague-Dawley rats aged 3 weeks (weanling [W]; n = 42), 8 weeks (young [Y]; n = 42), 16 weeks (teenage [T]; n = 39) and 56 weeks (old [O]; n = 40) were randomly assigned to groups: low fat, high sucrose and high fat with n = 12-16 per group. All animals were fed a purified low-fat, high starch diet for two weeks, and fed a low fat (STD), high sucrose (HSD), or high-fat (HFD) for five additional weeks. After sacrifice, the femurs were harvested and non-osseous tissue was removed. The bones were dried at 25 degrees C for 48 hours. Length and the periosteal minimum and maximum diameter (D-min and D-max) at the mid diaphysis of the femurs were measured with Vernier calipers. The femurs were rehydrated and tested via three-point flexure. Bones were weighed after drying at 105 degrees C (48 hours; Dry-M) and 800 degrees C (24 hours; Ash-M). Percent mineralization (%Min) was calculated as Ash-M/Dry-M X 100%. Length, D-min and D max, Dry-M and Ash-M all significantly (p < 0.05) increased with age (W < Y < T < O) within each group. %Min and stiffness were significantly greater in [O] than in the younger femurs. No significant results were seen in any age group due to varying diet. These results indicate that five weeks of high fat or high sucrose diet feeding does not affect whole bone size, composition or mechanical properties. Whether a longer dietary period or different diet composition would elicit changes requires further study. PMID- 10834264 TI - A study of purified montmorillonite intercalated with 5-fluorouracil as drug carrier. AB - In the recent decade, the most useful drug for the therapy of colon cancer is 5 Fluorouracil(5-FU). It has been reported to have considerable toxicity administered by intravenous injections or via alimentary treat. Though many materials have developed for drug carrier of 5-FU, there was no clinically acceptable carrier for 5-FU till now. Montmorillonite, one of clay minerals, consists of hydrated aluminum silicates that are fined grained and usually have a large space between the layers. Isomorphous substitution of cations is common. We try to intercalate 5-FU into interlayer of montmorillonite through ion-exchange. Montmorillonite with 5-FU intercalation is expected to achieve in-situ release for colon cancer therapy. In the study, 5-FU was dissolved in 100 ml distilled water as 5-FU solution. Purified montmorillonite powder will soak in 5-FU solution for a period of time ad different pH value and temperature. The intercalated amount of 5-FU in montmorillonite is measured by scanning differential thermal (SDT) analysis and UV analysis. The results showed that 4 g purified montmorillonite soaked in 0.6% 5-FU solution for 2 hours had an optimum condition for intercalation. The total amount of 5-FU in montmorillonite is about 9.13 wt%. PMID- 10834265 TI - Inertially compensated force plate: a means for quantifying subject's ground reaction forces in non-inertial conditions. AB - A system for the measurement of forces in noninertial reference systems, and a possible solution to compensate force plate readings by means of accelerometers were investigated. If the force plate can be considered a rigid body, six linear accelerometers can measure all linear and angular accelerations of the instrument in 3-D space. However, by using nine accelerometers in a proper layout, the force and moment generated by the movement of the plate can be quantified in a way that eliminates the errors due to the time integration of the accelerations. By doing so, no temporal limits are imposed to the compensation. A one degree of freedom numerical model was implemented to evaluate the importance of main error sources in the estimation of inertial forces using accelerometers with different technical characteristics. The inertial compensation of the force plate along one axis was tested, and the performances of a piezoelectric accelerometer and a capacitive accelerometer were compared. With the proposed inertially compensated force plate, accelerations up to 5 g's in amplitude and frequencies from 0 to 100 Hz can be compensated by means of capacitive accelerometers. Such a device can be used for force measurements in moving vehicles, or any situation where the surface on which the instrument is mounted is moving or vibrating. PMID- 10834266 TI - Accuracy of EEG dipole source localisation in presence of brain lesions. AB - Inverse solution techniques based on electroencephalographic (EEG) measurements are a powerful mean of gaining knowledge about brain functioning, being used to estimate location, orientation and strength of neural electrical sources of brain activity. A model of the head, a model of the source and an electric-field computational method are necessary to describe the EEG problem mathematically. Volume conductor models commonly used to describe the EEG neglect the presence of brain lesions. We evaluated the need of considering brain lesions in head models for precise mapping of neural activity nearby the lesion, as it is requested for neurosurgical preoperative planning. A systematic evaluation of the effects of neglecting brain lesions in EEG dipole source localisation accuracy has been performed by computer simulations for different pathologic conditions, source types and source positions. Simulations of EEG measurements were carried out using a modified eccentric-spheres model of the head in which an eccentric bubble approximates effects of actual brain lesions. A three concentric spheres model has been used in the inverse dipole fitting procedure to quantify source localisation errors caused by ignoring the presence of lesions in the head model. 64 different situations have been analysed. Source reconstruction errors resulted negligible only for some relative positions of source, lesion and electrodes. The largest errors, up to 2.5 cm, have been found for a lesion placed between source and electrodes and for a source internal to the lesion (not circumscribed tumours). We conclude that source localisation process is largely affected by a nearby lesion and thus electrical mapping of brain activity must be performed embedding brain lesions in the head model. PMID- 10834267 TI - Models of the uniformity of electro-magnetic fields generated for biological experiments by Merritt coils. AB - Electromagnetic field (EMF) producing wire coils were described by Merritt et al, Rev. Sci. Instrum. 54 (7), 1983. Merritt coils produce large volume EMFs in which statistical numbers of biological experiments are performed. We build and use Merritt coils for cell/animal studies and are developing therapeutic EMF systems. Here we present models illustrating the EMFs produced by our coils and discuss the criteria that should be applied to the use of Merritt and other coils to achieve valid experimental results. In a companion paper at this meeting Nindl et al, describe biological experiments, using these Merritt coils, showing that EMFs may be useful in treating many inflammatory disease states. Although the large volume EMFs produced by Merritt coils are convenient for biological experiments the EMFs are not perfectly uniform and the deviations can be a significant source of experimental error. The orientation and size of experimental objects are key contributors to these deviations. To evaluate our Merritt coils we solved the Biot-Savart law explicitly for ideal 3-coil and 4-coil Merritt systems and compared these theoretical EMFs with those of our systems. We present a detailed examination of deviations in magnetic field amplitude, as well as magnetic field direction, as a function of location within the coils. We find that spherically shaped experimental sets minimize these deviations. We developed simple formulae for accurately predicting deviations associated with Merritt coils. PMID- 10834268 TI - Virus-directed enzyme prodrug therapy using CB1954. AB - The virus-directed enzyme prodrug therapy (VDEPT) anti-cancer 'gene therapy' strategy relies on the use of viral vectors for the efficient delivery to tumour cells of a 'suicide gene' encoding an enzyme which converts a non-toxic prodrug to a cytotoxic agent. The prodrug 5-(aziridin-1-yl)-2,4 dinitrobenzamide, CB1954, has been proposed for use in enzyme-prodrug gene therapy systems with the Escherichia coli enzyme nitroreductase (Ntr). Ntr converts CB1954 to 2- and 4 hydroxylamino derivatives, whereupon the non-enzymatic reaction of the 4 hydroxylamino derivative with cellular thio- esters generates a potent cytotoxic bifunctional alkylating agent capable of cross-linking DNA. Ntr delivery has been achieved in vitro using retroviral and adenoviral vectors and confirmed by immunocytochemical demonstration of Ntr expression. The Ntr-expressing cells have been shown to be sensitized to CB1954 by up to 2000-fold. The Ntr-CB1954 system shows effective bystander killing in mixed populations of Ntr-expressing and non expressing cells treated with CB1954. The efficacy of this enzyme-prodrug approach in model systems compared with other VDEPT approaches demonstrates the feasibility and future promise of this gene therapy strategy. PMID- 10834269 TI - Antitumour prodrug development using cytochrome P450 (CYP) mediated activation. AB - An ideal cancer chemotherapeutic prodrug is completely inactive until metabolized by a tumour-specific enzyme, or by an enzyme that is only metabolically competent towards the prodrug under physiological conditions unique to the tumour. Human cancers, including colon, breast, lung, liver, kidney and prostate, are known to express cytochrome P450 (CYP) isoforms including 3A and 1A subfamily members. This raises the possibility that tumour CYP isoforms could be a focus for tumour specific prodrug activation. Several approaches are reviewed, including identification of prodrugs activated by tumour-specific polymorphic CYPs, use of CYP-gene directed enzyme prodrug therapy and CYPs acting as reductases in hypoxic tumour regions. The last approach is best exemplified by AQ4N, a chemotherapeutic prodrug that is bioreductively activated by CYP3A. This study shows that freshly isolated murine T50/80 mammary carcinoma and RIF-1 fibrosarcoma 4-electron reduces AQ4N to its cytotoxic metabolite, AQ4 (T50/80 Km = 26.7 microM, Vmax = 0.43 microM/mg protein/min; RIF-1 Km = 33.5 microM, Vmax = 0.42 microM/mg protein/min) via AQM, a mono-N-oxide intermediate (T50/80 Km = 37.5 microM; Vmax = 1.4 microM/mg protein/min; RIF-1 Km = 37.5 microM; Vmax = 1.2 microM/mg protein/ min). The prodrug conversion was dependent on NADPH and inhibited by air or carbon monoxide. Cyp3A mRNA and protein were both present in T50/80 carcinoma grown in vivo (RIF-1 not measured). Exposure of isolated tumour cells to anoxia (2 h) immediately after tumour excision increased cyp3A protein 2-3-fold over a 12 h period, after which time the cyp protein levels returned to the level found under aerobic conditions. Conversely, cyp3A mRNA expression showed an initial 3 fold decrease under both oxic and anoxic conditions; this returned to near basal levels after 8-24 h. These results suggest that cyp3A protein is stabilized in the absence of air, despite a decrease in cyp3A mRNA. Such a 'stabilization factor' may decrease cyp3A protein turnover without affecting the translation efficiency of cyp3A mRNA. Confirmation of the CYP activation of AQ4N bioreduction was shown with human lymphoblastoid cell microsomes transfected with CYP3A4, but not those transfected with CYP2B6 or cytochrome P450 reductase. AQ4N is also reduced to AQ4 in NADPH-fortified human renal cell carcinoma (Km = 4 microM, Vmax = 3.5 pmol/mg protein/min) and normal kidney (Km = 4 microM, Vmax = 4.0 pmol/mg protein/min), both previously shown to express CYP3A. Germane to the clinical potential of AQ4N is that although both normal and tumour cells are capable of reducing AQ4N to its cytotoxic species, the process requires low oxygen conditions. Hence, AQ4N metabolism should be restricted to hypoxic tumour cells. The isoform selectivity of AQ4N reduction, in addition to its air sensitivity, indicates that AQ4N haem coordination and subsequent oxygen atom transfer from the active-site-bound AQ4N is the likely mechanism of N-oxide reduction. The apparent increase in CYP3A expression under hypoxia makes this a particularly interesting application of CYPs for tumour-specific prodrug activation. PMID- 10834270 TI - Nitracrine N-oxides: effects of variations in the nature of the side chain N oxide on hypoxia-selective cytotoxicity. AB - The tertiary amine N-oxide (nitracrine-N-oxide, 1b) of the 1-nitroacridine nitracrine is a bis-bioreductive agent showing very high hypoxic selectivity (approximately 1000-fold) against tumour cells in culture, but only modest activity against the hypoxic subfraction of tumours in vivo. Because the hypoxic selectivity of 1b was considered to depend significantly on the rate of enzyme mediated reduction of the N-oxide group, this paper reports the preparation and evaluation of a series of analogues in which the environment of this group was modified. Three analogues contained more weakly basic N-oxides, while two others had varying degrees of steric bulk around the N-oxide. In all but one case (an aromatic N-oxide), the N-oxides were much less cytotoxic (10- to 300-fold) than the corresponding tertiary amines towards AA8 Chinese hamster cells under aerobic conditions. Both the N-oxides and the corresponding amines were more cytotoxic to an ERCC-1 mutant defective in nucleotide excision repair, indicating that DNA alkylation was the cytotoxic event. However, there was no apparent correlation of these parameters with structure. All of the aliphatic N-oxides, with the exception of the aromatic N-oxide example, showed substantial (70- to 800-fold) hypoxic selectivity against AA8 cells in a clonogenic assay. While the weakly basic derivatives were the least selective, there was no apparent relationship between hypoxic selectivity and the steric environment of the N-oxide. Selectivity for hypoxic cells in culture is shown to depend on the hypoxic selectivity of the corresponding tertiary amine (reflecting O2-inhibitable reduction of the 1-nitro group) and the differential in aerobic toxicity between amine and N-oxide (a measure of the potential toxicity increase achievable by reducing the N-oxide). Four analogues whose structures fairly represented the range of steric and electronic modifications of the N-oxide site were evaluated against the hypoxic subfraction of cells in KHT tumours in vivo, but were inactive. These results suggest that either such modifications do not exert significant effects on N-oxide reduction, or that the rate of such reduction is not a factor limiting the in vivo activity of the parent analogue 1b. PMID- 10834271 TI - Camptothecin delivery systems: the utility of amino acid spacers for the conjugation of camptothecin with polyethylene glycol to create prodrugs. AB - The primary purpose of this study was to screen individual amino acid spacers in polyethylene glycol (PEG) conjugated camptothecin for their impact on the conjugates' antitumor activity. Secondly, an active member of this series was used to assess the PEG-camptothecin conjugate's efficacy against a battery of solid tumor types. PEG-camptothecin is a novel water soluble transport form (macromolecular prodrug) of the naturally derived antitumor drug, 20-(S) camptothecin (CPT). Rates of hydrolysis were studied in phosphate buffered saline (PBS) and the plasma of both rats and humans. In vivo efficacy screens were performed against P388/0 murine leukemia and LS174T human colon solid tumor xenograft models. The results showed that while all the derivatives had considerable stability in PBS, their rates of hydrolysis varied in both rat and human plasma according to the amino acid spacer employed. Not surprisingly, changing the amino acid also affected in vivo toxicity and efficacy in the treatment of ascites and solid tumors. A representative of this amino acid series, PEG-alanine-CPT, which showed moderate activity in the solid tumor screen, was chosen for evaluation of efficacy across a wide range of solid tumor types and demonstrated significant antitumor activity (% T/C < 30%) in all tested xenograft models (colon, ovarian, mammary, lung, pancreatic and prostate). Therefore, this study showed that the use of specific amino acid spacers affected both the PEG-camptothecin conjugates' breakdown and biological activity. We anticipate that using these insights, this soluble macromolecular transport technology could be successfully employed with a number of antitumor drugs. PMID- 10834272 TI - Intensely cytotoxic anthracycline prodrugs: galactosides. AB - We have reported the synthesis of a series of anthracycline analog prodrugs that give rise to intensely cytotoxic metabolites in the presence of carboxylate esterases and beta-glucuronidases. We now report structurally related prodrugs that are converted to similar potent metabolites in the presence of beta galactosidases. The prototypical compound, N-[(4"RS)-4"-ethoxy-4"(1'"-O-beta-D galactopyranosyl)butyl]daunorubicin, 8a, was prepared by reductive condensation of daunomycin with 1-O-[(1'RS)-1'-ethoxy-4'-oxobutyl]-2, 3, 4, 6-tetra-O-acetyl beta-D-galactopyranoside in the presence of sodium cyanoborohydride, followed by deacetylation of the galactoside moiety with sodium methoxide. A related prodrug (8b) with enhanced lipophilicity (the 4'-hexoxy analog of 8a) and 8c (the propyldaunomycin analog of 8a) were prepared for comparative studies. 8a and 8b were isolated after chromatography on silica as a mixture of 4'R and 4'S diastereomers; 8c, on the other hand, was resolved into its component 3' diastereomers, 8c(R) and 8c(S). 8a, 8c(R) and 8c(S) showed no evidence of decomposition when incubated at 37 degrees C in 0.05 M phosphate buffer, pH 7.4, for 2 weeks; 8b, under the same conditions, was degraded with a half-life of 49 h. In the presence of two units of Escherichia coli beta-galactosidase per pmol of substrate, the half-lives of 8a, 8b, 8c(R) and 8c(S) were 1.98, 1.06, 3.5 and 2.4 h, respectively. HPLC analysis of the incubation mixtures showed that 8a and 8b gave rise to a single, chromatographically identical metabolite. 8c(R) and 8c(S) also gave rise to a single, identical metabolite. 8a and 8b were nearly one million-fold more toxic to human A375 melanoma cells in culture in the presence of E. coli beta-galactosidase than in the absence of the enzyme. The activation products of 8c(R) and 8c(S) were approximately 1000-fold less potent. These beta galactoside prodrugs have chemotherapeutic potential for use in conjunction with tissue-targeting strategies such as antibody-directed enzyme prodrug therapy (ADEPT) and gene-directed enzyme prodrug therapy (GDEPT). PMID- 10834273 TI - Prodrugs for antibody- and gene-directed enzyme prodrug therapies (ADEPT and GDEPT). AB - Antibody- and gene-directed enzyme prodrug therapy are two-step targeting strategies designed to improve the selectivity of antitumour agents. The approaches are based on the activation of specially designed prodrugs by antibody enzyme conjugates targeted to tumour-associated antigens (ADEPT) or by enzymes expressed by exogenous genes in tumour cells (GDEPT). Herein the design, synthesis, physico-chemical and biological properties, kinetics and clinical trials of the prodrugs and the enzymes carboxypeptidase G2 and nitroreductase are reviewed for ADEPT and GDEPT. PMID- 10834274 TI - Synthesis and antitumor activities of amino acid prodrugs of amino combretastatins. AB - The synthesis and antitumor activity of water-soluble amino acid prodrugs of amino-combretastatins were reported. Among the synthesized compounds, 7e (CS-39-L Ser HCI, AC-7700) showed enhanced antitumor activity and decreased toxicity in a Colon 26 murine adenocarcinoma model. Compound 7e showed improved solubility and was easily formulated for in vivo administration. Compound 7e was cleaved to generate the parent compound, CS-39, in the whole blood of mice as well as man, possibly by the action of amino peptidase on the erythrocyte membrane. PMID- 10834275 TI - Future directions in health for the new Government: our health and the public health. AB - The 'no model' model awaited the invisible hand. However, 'more government' demands more hands-on direction of the health system. To whom will the government turn for advice? The health professionals were excluded from the market experiment because of the concern that they would 'capture the process'. With the change in the political will, some of the most simplistic, fallacious perceptions of New Zealand's society should have less influence on health policy. The new government has the opportunity to re-engage New Zealanders in advancing the health of the nation. Public health specialists have a wealth of information to guide decision-making which deserves wider dissemination and more executive consideration. In personal health care, health professionals have the will to continue innovation but need a system in which their values are respected. One caveat: there is a tendency, whether the approach is 'more market' or 'more government', to try to measure everything and tick every box. When dealing with complex systems which need highly skilled professionals, however, there simply needs to be a high level of trust As when getting on a plane, central planners, like the rest of us, need to understand that sometimes the pilots must 'capture the process' in order to get back on course. PMID- 10834276 TI - Managerialism in public hospitals and universities in New Zealand. PMID- 10834277 TI - Liver function and hepatitis markers in carriers of hepatitis B virus in New Zealand. AB - AIMS: The Hepatitis Foundation has identified many chronic carriers of hepatitis B virus (HBV) in community surveys of schools and family contacts. This study reports the characteristics of carriers and the relationship between hepatitis markers and liver function. METHODS: Demographic data from confirmed chronic carriers of HBV in the North Island were correlated with liver function and hepatitis markers. Longitudinal data were obtained by following a cohort for two years with regular blood tests. RESULTS: Of 2778 confirmed carriers of HBV most were children or young adults and 56% were male. Sixty percent were Maori and 26% Pacific Island people. Loss of HBsAg occurred at less than 1% per year compared to 9% for HBeAg. Mean ages for 50% loss of HBeAg were 14 years for children of HBV negative mothers and 19 years for those of HBV carrier mothers. Fewer adult males than females were HBeAg positive. Alanine aminotransferase levels above 50 IU/L were found in 16% of HBeAg positive and 6% HBeAg negative cases. Other factors significantly associated with raised alanine aminotransferase were male gender (OR 1.8) and age more than 15 years (OR 2.0). Thirty five percent of HBeAg positive carriers with raised alanine aminotransferase levels spontaneously seroconverted to HBeAg negative in two years. However, raised alanine aminotransferase in HBeAg negative carriers was persistent in most cases and 38% had HBV-DNA detectable in serum. CONCLUSIONS: HBV carriage is less benign in adults than children, even after loss of HBeAg. It is recommended that all HBV carriers have regular checks of liver function. Those with persistent abnormality should be strongly advised to restrict alcohol and be assessed for possible antiviral treatment. PMID- 10834278 TI - Establishment of a smoking cessation programme in primary and secondary care in Canterbury. AB - AIMS: Smokescreen for the 1990's is a smoking cessation programme devised for use in primary care in Australia. It is based on the 'readiness to change' model where smokers are categorised as being 'ready', 'unsure', or 'not ready' to quit smoking. Those in the 'ready' group are encouraged to set a quit date and offered nicotine replacement therapy. Those in the unsure group receive brief motivational intervention, and those 'not ready' are given simple health advice. The aims of the study were to evaluate the process and outcome of establishing this program in primary and secondary care in Christchurch. METHODS: Process evaluation involved all staff participating in the program. Patient outcomes including quit rates were assessed by interview six months after enrollment. RESULTS: Implementation was successful with 59 general practitioners, 49 practice nurses and 294 hospital staff receiving education in the use of the programme. Nine hundred and seven patients (smokers) were enrolled in the study, 347 from primary care and 560 from Christchurch Hospital. The point prevalence abstinence rate at 6 months was 10.4% for the primary care sample and 17% for the secondary care group, with an overall rate of 14.4%. CONCLUSIONS: The programme was successfully implemented across primary and secondary care with an acceptable quit rate at 6 months. PMID- 10834279 TI - Stroke rehabilitation for the older person: current evidence, potential problems and future challenges. PMID- 10834280 TI - Lost in the smoke: tobacco control in New Zealand during the 1990s. PMID- 10834281 TI - Geographic perspectives on hospital restructuring and its impacts in New Zealand. PMID- 10834282 TI - Addressing concerns. PMID- 10834283 TI - Diabetes awareness. PMID- 10834284 TI - "Fantasy"-drug of abuse. PMID- 10834285 TI - Occupationally acquired tuberculosis in New Zealand. PMID- 10834286 TI - Doctor guilty of disgraceful conduct. PMID- 10834287 TI - Biological adaptation and social behaviour. AB - In 1930, both Fisher and Wright identified Darwin's initial concept of adaptive evolution in the light of the genetical theory with intergenerational variation in allelic frequencies brought about by the action of natural selection through differential reproduction. They emphasized that selection only works at the level of the individual and that its only consequence is to increase fitness. One genetical evolution not easy to explain on these bases was that of social behaviour because any altruistic gene disadvantageous for its carriers in an antisocial environment would have been opposed by selection. In the 1950s, ethologists focusing on what appeared to be evolved collective behaviours, hypothesized that selection could operate at group level. Though the controversy between group selectionists and evolutionary geneticists ended by the rejection of the evolutionary role of group selection, it has remained a subject of investigation until now. Kin selection, proposed by Hamilton, offered a solution to the problem of the evolution of altruism and gave the impetus to the trend of adaptive explanations of basic behaviours, which was to become the core of human sociobiology. The intrusion of behaviour into the process of adaptive evolution was an invitation to investigate culture as an evolutive process. The first sociobiological interpretations of culture as a derivative of genetic processes were followed by other ideas in which culture, though channelled by evolved predispositions, was essentially free from biological determinism. It is concluded that as we have come to better understand human adaptation, its complexities have been further revealed, a development already implicit in Darwin's notion. PMID- 10834288 TI - Adiposity indices and their relationship with some risk factors of coronary heart disease in middle-aged Cambridge men and women. AB - The relationship between five anthropometric measures (body mass index, waist to hip ratio, conicity index, waist circumference and waist to height ratio) and seven cardiovascular risk factors (total cholesterol, high and low density lipoprotein cholesterol, ratio of total cholesterol to high density lipoprotein and ratio of high density lipoprotein to low density lipoprotein cholesterol, systolic and diastolic blood pressure), were studied in 165 men and 202 women aged between 40 and 69 years living in an urban area of Cambridge, UK. Inter correlations between the anthropometric measures varied between 0.34 and 0.92 and were generally higher in males than females. Tests of homogeneity of the five correlation coefficients for each risk factor revealed no significant heterogeneity for males, but significant differences for high density lipoprotein and ratio of total cholesterol to high density lipoprotein were found in females. Waist to hip ratio showed the highest correlations with the risk factors in women (3 out of 7), but no consistent pattern was found for males. Males had significantly lower cholesterol, low density lipoprotein cholesterol levels than females for any given waist to hip ratio and conicity index, waist circumference and waist to height ratio. In addition, men also had lower total cholesterol to high density lipoprotein cholesterol and high density lipoprotein cholesterol to low density lipoprotein cholesterol levels for waist to hip ratio or conicity index. PMID- 10834289 TI - Nutritional status and age at menarche in a rural area of Bangladesh. AB - The age at menarche and its association with nutritional status in a rural area of Bangladesh was determined. A cross-sectional study was conducted in four villages of Rupganj Thana of Narayanganj district. Data was collected through October to December 1996 using a pre-tested structured questionnaire interview schedule, and nutritional status was measured by weight, height, body mass index (BMI) and physical examination. Data were obtained on 436 adolescent girls aged 10-17 years. Among them, 165 (37.8%) girls had commenced menarche. The mean age at menarche as determined by retrospective recall was 13 years SD 0.89 (n = 165). The median age at menarche determined by the status quo method was 13.0. Among the adolescents 60.1% were thin (BMI < 5th centile WHO recommended reference) and 48.2% were stunted (< 3rd centile NCHS/WHO). The mean weight and BMI were significantly higher among the menstruating girls of 13, 14 and 15 years (p < 0.01) than non-menstruating girls. The mean height was found to be significantly higher at 11-14 years among the menstruating girls (p < 0.05). A lower prevalence of angular stomatitis was found among the menstruating adolescent girls compared with the non-menstruating girls, 36.4% versus 46.5%, although this was statistically non-significant (odds ratio = 0.66, 95% CI 0.43-1.00). For glossitis, no significant difference was found. Among the menstruating girls 12.1% were suffering from menorrhagia and 31.5% from dysmenorrhoea. We conclude that the age of menarche among this rural Bangladeshi community is not as delayed as expected. Not surprisingly, menarche is associated with better nutritional status. The surveyed population had extremely high rates of undernutrition which suggests that adolescents in this and similar situations require specific intervention programmes to improve their nutritional status. PMID- 10834290 TI - Anthropometric survey of the elderly in south-western Nigeria. AB - One hundred and ninety-four subjects aged 65-78 years from rural and urban areas of the south-western region of Nigeria have been surveyed for height, weight, upper arm, hip and waist circumferences. The 24-hour dietary recall technique was also employed to assess their dietary energy intake. In both rural and urban cohorts, male subjects were significantly taller and weighed more (p < 0.05) than female subjects. There were no significant differences in the height of rural groups and their respective urban groups, although urban males and females weighed significantly (p < 0.05) more than their respective rural counterparts. Mean body mass index (BMI) ranged from 18.4 to 21.1 kg m(-2), and 73% of all subjects had a BMI below 20% and 10% were below 18.5. Waist, hip and upper arm circumferences of urban cohorts consistently exceeded those of rural subjects, although only for females were these differences statistically (p < 0.05) significant. Significant differences observed in the energy intake (per kg body weight) are offered as one explanation for the superior anthropometric indices of urban as compared with rural elderly in Nigeria. PMID- 10834291 TI - Secular trends in height, weight and body mass index of 6-year-old children in Bremerhaven. AB - Secular trends in growth processes of children can be important indicators of changes in public health. Common to studies on secular trends in children is that evaluation is based on comparison of data collected at two (or more) distinct points on a time scale. The quantitative characteristic of the secular trend is estimated by linear interpolation between the two end points of the underlying time interval, which in studies of children are usually at least 10 years apart. The purpose of the present paper is to analyse secular trends in height, weight and body mass index (BMI) of 6-year-old children from Bremerhaven over the period 1968-1987 (the year refers to the birth cohort). The results are based on data drawn from health records of the City Health Centre, where all 6-year-old children are routinely measured in a school entrance examination. Thus the data represent complete birth cohorts of children entering school in Bremerhaven and not selected samples. The data reported here refer only to children of German origin. The sample sizes vary from n = 313 (girls born in 1982) to n = 737 (boys born in 1968), and total sample size is n = 7601. Regression of the arithmetic means of height on year of birth showed that the trend in stature for children born between 1968 and 1987 was 0.67 cm/decade for boys and 0.49 cm/decade in girls. Both trends are statistically significant (p < 0.05). Although there was an increasing tendency for weight as well, which was more marked for the 95th percentile than for the median, neither of the trends in both sexes was statistically significant. While the BMI in both sexes showed no trend at all for the median and the 5th percentile, there was a significant linear increase of the 95th percentile. Furthermore, the results for height show that an evaluation of secular trends under qualitative and quantitative perspective critically depends on the selection of points on the time scale. PMID- 10834292 TI - Plasma leptin and insulin levels in Aymara natives from Chile. AB - The purpose of this study was to examine the relationship of plasma leptin levels with respect to obesity, gender, age and insulin levels in the native Aymara population. The Aymara natives live at high altitudes in isolated regions in the north of Chile, and they maintain distinctive genetic and cultural characteristics. Plasma leptin and insulin levels were correlated with body mass index (BMI), sex and age in a sample of 147 adult Aymara subjects who participated in a cross-sectional study. Multivariate analysis showed significant differences in leptin levels (dependent variable: natural log of leptin) by gender (p < 0.0001), and by BMI (p < 0.001), without significant statistical interaction between gender and BMI. The effect of age achieved statistical significance in the multivariate analysis (p = 0.02). Gender, BMI and insulin are independently associated with plasma leptin levels. On the other hand, the multivariate analysis of the plasma insulin concentration (dependent variable: natural log of insulin levels) shows that insulin is strongly associated with BMI (p < 0.0001), although non-statistically significant differences of insulin levels by sex (p = 0.07) or age (p = 0.9) were detected at alpha 0.05 level. Thus, in the special ecosystem where the Aymara population live, a strong and independent association between sex, obesity and insulin levels with plasma leptin levels has been detected. PMID- 10834293 TI - Contribution of growth phases to adult size. AB - Based on the data of the First Zurich Longitudinal Growth Study we investigate how interindividual differences in adult size arise in the variables leg height, sitting height and standing height, arm length, bi-iliac width and bihumeral width. Specifically, we are also interested in the question of whether across sexes and variables the same growth phases and the same parameters are predictive for achieving a certain adult size. A rather complex pattern emerges, demonstrating that regulation of growth is not the same for boys and girls and moreover is not the same for the six anthropometric variables studied. Prepubertal growth is characterized by its intensity (average velocity) and by its duration. Whereas duration has by itself no appreciable influence on adult size, prepubertal intensity determines adult size to a high degree across all variables and both sexes. The intensity of prepubertal growth determines adult size to a larger degree for boys than for girls. For a given size at the end of the prepubertal period, a small duration enhances the chance of obtaining a large adult size. Compared with prepubertal growth, the amount of variance of adult size explained is small for pubertal parameters, and--with respect to linear measures--significant for girls only. A small duration of prepubertal growth is in the following mainly compensated by a stronger pubertal spurt (PS), to a varying degree across variables. The overall picture which emerges indicates that sitting height--and to a lesser extent bihumeral width--develop in a more irregular fashion than the variables bi-iliac width and leg height. PMID- 10834294 TI - Genetic analysis of growth curve parameters of body weight, height and head circumference. AB - A sample of 681 Israeli boys and girls, including 355 regular siblings (SB), 112 pairs of dizygotic (DZ) and 51 pairs of monozygotic (MZ) twins, was measured for body weight (WT), length (HT) and head circumference (HC) at birth and during the first year of life. The Count model with three parameters was chosen as the best fitting and most parsimonious function to approximate growth of the studied traits. The curves' fitting parameters were estimated for WT, HT and HC for each individual. To test the assumption that there is a genetic source influencing the pattern of growth for each trait, familial correlations between parameter estimates were computed for MZ, DZ twins and SB. In all instances MZ twins showed the highest within-pair correlation in parameters of growth (from 0.58 to 0.86), while SB showed the lowest ones (from 0.10 to 0.70). Variance decomposition analysis was used to simultaneously assess the contribution of gender, gestational age, additive genetic factor, common sibs and common intrauterine environmental effects on total variance of each studied trait separately. All these sources of variation were statistically significant, though the effect of intrauterine environment played a substantial role in early stages of child physical development, explaining from 18.1% to 70.6% of the total variance of the growth curve parameters. Further analyses are needed to clarify how this environment affects child growth and for how long. PMID- 10834295 TI - Trend in menarcheal age in Spain: rural and urban comparison during a recent period. AB - Menarcheal age of a sample of rural (n = 342) and urban (n = 469) girls with ages ranging from 9 to 16 years from Madrid province was analysed. Results, obtained through the status quo method and probit analysis (age 12.79 SD 1.04 in Madrid city and 12.90 SD 1.03 in rural sample), were compared with data from previous studies carried out in the same area, as well as those from other regions of the Iberian Peninsula. The aim of these comparisons was to establish the variability of menarche in the recent past, considering physical environment, degree of urbanization and socio-economic development. Decline in menarcheal age was detected and, in general, a tendency towards similar means among Spanish populations in the period studied. PMID- 10834296 TI - A study of consanguinity in the Sultanate of Oman. AB - Omani society has a long tradition of consanguinity, which goes back to pre Islamic times. The aim of the present study was to determine the prevalence of consanguineous marriage and the mean coefficient of inbreeding in Oman. Consanguinity rates among 60,635 couples (20.8% of the national population of childbearing age) were determined on the basis of a questionnaire in major delivery units. In this large survey, which included all sections of the community, 24.1% of marriages were reported between first cousins and 11.8% of marriages were between second cousins. In addition, a further 20.4% of marriages were contracted within specific tribal groupings. Because of the strictly endogamous nature of the tribal groups in Oman, all marriages would be expected to be consanguineous to some degree, albeit at a level beyond that of second cousins (F < or = 0.0156). To obtain a more detailed picture of the patterns of consanguinity, and to assess the validity of the questionnaire-based study, 500 pedigrees were investigated in detail. The mean coefficient of inbreeding (alpha) in these pedigrees was 0.0204, compared with 0.0198 in the larger survey, suggesting that the data were broadly comparable and confirming the high degree of consanguinity in the population of Oman. PMID- 10834298 TI - Zinc may be a mediator of leptin production in humans. AB - Obese individuals have hyperleptinemia and hypozincemia. Moreover, leptin and zinc have circadian changes in circulating concentrations. We investigated their possible interaction and examined whether a difference existed between obese men and their lean controls. The results indicated the pattern of circadian change in plasma zinc and leptin did not markedly differ between the obese subjects and the lean controls. However, the obese had higher leptin and lower zinc plasma values at each sampling time than did the lean controls. Because an inverse correlation was found in plasma values between zinc and leptin (r=-0.51, p=0.012), we further determined the role zinc might play in leptin production by human subcutaneous adipose tissue from female donors. The in vitro study showed that zinc treatment (0.2 mmol/L) significantly increased leptin production (142%), however, this increment did not surpass that by insulin (10 nmol/L). The data of this study suggest an interactive connection between zinc and leptin. PMID- 10834297 TI - Chlorinated dibenzo-p-dioxins and dibenzofurans and the human immune system: 3. Plasma immunoglobulins and cytokines of workers with quantified moderately increased body burdens. AB - The concentrations of immunoglobulins (IgA, IgD, IgG, IgM) and of several cytokines were measured in the plasma of volunteers with clearly, but moderately, increased body burdens of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/PCDF), using monoclonal antibodies and an enzyme-linked immuno-sorbant assay. Two groups of workers with different body burdens of PCDD/PCDF were studied: (trial I) persons with mainly 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), and (trial II) persons with mainly penta- and hexachlorinated dibenzofurans (P5CDF/H6CDF) in their blood fat. Including the reference group, 158 volunteers were investigated. A slight but statistically significant decrease was observed in the plasma concentration of IgG1 in persons exposed to TCDD, but not in persons exposed to P5CDF/H6CDF. When the data of both groups were pooled and a multi-regression analysis against international TCDD toxicity equivalencies (I-TEq, NATO/CCMS) was performed, taking several confounding factors into account, no influence of the dioxin exposure could be revealed. There were no changes in the plasma concentrations of the other immunoglobulins studied. In the same volunteers, no deviation from the reference range was found for the concentrations of the cytokines: IL-1alpha, IL-1beta, IL-6 and TNFalpha in blood plasma. PMID- 10834299 TI - Inhibitory effects of sesquiterpenes from bay leaf on nitric oxide production in lipopolysaccharide-activated macrophages: structure requirement and role of heat shock protein induction. AB - The methanolic extract from the leaves of Laurus nobilis (bay leaf, laurel) was found to inhibit nitric oxide (NO) production in lipopolysaccharide (LPS) activated mouse peritoneal macrophages. Through bioassay-guided separation, fourteen known sesquiterpenes were isolated from the active fraction and were examined for ability to inhibit the NO production. Seven sesquiterpene lactones (costunolide, dehydrocostus lactone, eremanthine, zaluzanin C, magnolialide, santamarine and spirafolide) potently inhibited LPS-induced NO production (IC50 = 1.2 approximately 3.8 microM). Other sesquiterpene constituents also showed the inhibitory activity (IC50 > or = 21 microM), but their inhibitory activities were less than those of sesquiterpene lactones. Alpha-methylene-gamma-butyrolactone also showed inhibitory activity (IC50 = 9.6 microM), while mokko lactone and watsonol A etc., reductants of the alpha-methylene-gamma-butyrolactone moiety by NaBH4 or DIBAL, and a 2-mercaptoethanol adduct of dehydrocostus lactone showed little activity (IC50 > or = 18 microM). These results indicated that the alpha methylene-gamma-butyrolactone moiety is important for the activity. Furthermore, costunolide and dehydrocostus lactone inhibited inducible nitric oxide synthase (iNOS) induction in accordance with induction of heat shock protein 72 (HSP 72). These results suggested that, as one of their mechanisms of action, sesquiterpene lactones induce HSP 72 thereby preventing nuclear factor-kappaB activation followed by iNOS induction. PMID- 10834300 TI - Effects of dopamine metabolites on locomotor activities and on the binding of dopamine: relevance to the side effects of L-dopa. AB - L-dopa is the major treatment for Parkinson's disease (PD), but its efficacy is limited by the presence of dyskinesia. The dyskinesia develops over a period of exposure to L-dopa and is related to the dosage, therefore, the cause may involve inductive changes that produce toxic levels of metabolites, interfering with dopamine (DA) neurotransmission. Chronic L-dopa induces catechol-O methyltransferase (COMT) and methionine adenosyl transferase (MAT), enzymes involved in the methylation of catecholamines (CA). In addition, high levels of 3 O-methyl-dopa have been reported in the plasma of dyskinetic PD patients, treated with L-dopa, as compared to non-dyskinetic patients, therefore, the methyl metabolites of CA may be increased during L-dopa therapy and may be involved in the dyskinesia. Since large amounts of DA are produced from L-dopa, and DA is extensively methylated, the methyl metabolites of DA, 3-methoxytyramine (3-MT) and 3,4-dimethoxyphenylethylamine (DIMPEA), may be also involved. The first step in knowing this, is to assess the behavioral and DA-receptor activities of 3-MT and DIMPEA. In the rat, the intraventricular injection of 0.5 micromol of DIMPEA increased the total distance traveled (TD) by over 100%, the number of movement (NM) made by 40% and the time spent moving (MT) by about 36%. Identical doses of 3-MT decreased the TD by 42%, NM by 22% and MT by 39%. DIMPEA (1 mM) increased the binding of DA with brain membranes by 44.7%, whereas 3-MT decreased it by 15.8%. The results show that 3-MT and DIMPEA are behaviorally active, and in parallel, they interact with the binding sites for DA, consequently, they may contribute to the side effects of L-dopa. L-dopa produces high levels of DA and induces MAT and COMT. It is proposed, therefore, that DA will be methylated to 3 MT and 3-MT to DIMPEA. At threshold level each product will inhibit, allosterically, its enzyme of methylation, causing sequential and rhythmic up and down regulation of its concentration. At peak levels these hydrophobic metabolites will modulate the actions of DA on synaptic membranes, causing abnormal movements, at times, resembling the "on-off effects". PMID- 10834301 TI - Inhibitory effects of PGD2, PGJ2 and 15-deoxy-delta12,14-PGJ2 on iNOS induction in rat mesenteric artery. AB - PGD2 and its metabolites PGJ2 and 15-deoxy-delta12,14-PGJ2 have been reported to inhibit iNOS induction in cultured vascular smooth muscle cells. The present study was undertaken to determine whether these prostanoids inhibit iNOS induction in the isolated rat mesenteric artery. The artery without endothelium was incubated with and without lipopolysaccharide (LPS) at 37 degrees C for 6 hrs, then washed and mounted in an organ bath to measure isometric changes in tension. L-arginine but not D-arginine (10(-6) - 10(-3) M) induced concentration dependent relaxations only in the artery preincubated with LPS, the relaxations of which were attenuated by L-N(G)-nitroarginine methyl ester (LNAME, 10(-4) M), a non-selective iNOS inhibitor, and 1400W (10(-5) and 10(-4) M), a selective iNOS inhibitor. Co-treatment of cycloheximide (10(-5) M), a protein synthesis inhibitor, or actinomycin D (10(-7) M), an RNA synthesis inhibitor with LPS inhibited the development of relaxing ability in response to L-arginine, indicating iNOS induction by LPS. PGD2, PGJ2 and 15-deoxy-delta12,14-PGJ2 but not PGE2, PGI2 or PGF2alpha also inhibited the development of relaxing ability in response to L-arginine when added during incubation with LPS. Incubation of the artery with LPS at 37 degrees C for 6 hrs markedly increased production of nitric oxide (NO), which was abolished by 15-deoxy-delta12,14-PGJ2 (10(-5) M). An imunohistochemical study using antibody against murine iNOS showed that 15-deoxy delta12,14-PGJ2 (10(-5) M) inhibited the expression of iNOS protein in isolated rat mesenteric arteries. These results demonstrated that PGD2 and its metabolites inhibit iNOS induction by LPS in isolated rat mesenteric arteries, resulting in reduced relaxing ability in response to L-arginine. PMID- 10834302 TI - Molecular cloning of Rhombex-40 a transmembrane protein from the ventral medullary surface of the rat brain by differential display. AB - Respiration-related neurons, which detect various chemicals in cerebrospinal fluid, are localized to the ventral medullary surface (VMS). We hypothesized that expression of genes involved in respiratory function is upregulated in the VMS. By differential display, we looked for genes differentially expressed in VMS neurons and cerebral cortical neurons. Seventeen clones of interest were isolated, and sequence analysis revealed that one of these clones encoded a putative transmembrane protein, rhombencephalic expression protein-40 kDa (Rhombex-40). The rat Rhombex-40 was composed of 374 amino acid residues, and the predicted secondary structure displays a signal peptide in the N-terminus and single-pass transmembrane domain in the center of the sequence. An analysis of consensus sequences identified several phosphorylation sites in the intracellular domain. Expression of rat Rhombex-40 mRNA is high in the brain, and low in lung, liver and kidney. No homologous protein sequence was found in database searches. Whereas the biological function of this protein is presently unknown, its structural features and high expression in the brain suggest that Rhombex-40 may function as a novel transmembrane molecule in neural cells of the brain. PMID- 10834303 TI - Role of cytochrome P450 3A in the metabolism of mefloquine in human and animal hepatocytes. AB - We studied mefloquine metabolism in cells and microsomes isolated from human and animal (monkey, dog, rat) livers. In both hepatocytes and microsomes, mefloquine underwent conversion to two major metabolites, carboxymefloquine and hydroxymefloquine. In human cells and microsomes these metabolites only were formed, as already demonstrated in vivo, while in other species several unidentified metabolites were also detected. After a 48 hr incubation with human and rat hepatocytes, metabolites accounted for 55-65% of the initial drug concentration, whereas in monkey and dog hepatocytes, mefloquine was entirely metabolized after 15 and 39 hrs, respectively. The consumption of mefloquine was less extensive in microsomes, and unchanged drug represented 60% (monkey) to 85 100% (human, dog, rat) of the total radioactivity after 5 hr incubations. The involvement of the cytochrome P450 3A subfamily in mefloquine biotransformation was suggested by several lines of evidence. Firstly, mefloquine metabolism was strongly increased in hepatic microsomes from dexamethasone-pretreated rats, and also in human and rat hepatocytes after prior treatment with a cytochrome P450 3A inducer. Secondly, mefloquine biotransformation in rifampycin-induced human hepatocytes was inhibited in a concentration-dependent manner by the cytochrome P450 3A inhibitor ketoconazole and thirdly, a strong correlation was found between erythromycin-N-demethylase activity (mediated by cytochrome P450 3A) and mefloquine metabolism in human microsomes (r=0.81, P < 0.05, N=13). Collectively, these findings concerning the role of cytochrome P450 3A in mefloquine metabolism may have important in vivo consequences especially with regard to the choice of agents used in multidrug antimalarial regimens. PMID- 10834304 TI - Long-term treatment with SR141716A, the CB1 receptor antagonist, influences morphine withdrawal syndrome. AB - The role of the cannabinoid system in morphine withdrawal was examined through long-term CB1 receptor antagonist administration in morphine pellet implanted rats. SR141716A chronic treatment (5mg/kg i.p. twice a day for four days) did not influence the development of tolerance to the morphine analgesic effect but significantly reduced the intensity of naloxone-induced opiate withdrawal in tolerant rats: Specifically there was a significant reduction in the number of digging, teeth chattering and penile licking and the incidence of diarrhoea while other signs such as writhing, head dog shakes and rearing were unaffected. These results suggest that the pharmacological treatment with SR141716A could be of some interest in ameliorating opiate withdrawal syndrome. PMID- 10834305 TI - Thioperamide, a selective histamine H3 receptor antagonist, protects against PTZ induced seizures in mice. AB - The effect of selective histamine H3-receptor antagonist thioperamide was studied on PTZ-induced seizures in mice. Thioperamide significantly protected clonic seizures induced by PTZ in a dose-dependent manner. The effect of thioperamide was completely countered by pretreatment with R (alpha)-methylhistamine (RAMH), a selective H3-receptor agonist suggesting that the observed effect of thioperamide was elicited by histamine H3-receptors. RAMH alone did not significantly modify PTZ seizures. The findings are consistent with a role for the histaminergic neuronal system in seizures and suggest that H3-receptors may play an important role in modulating clonic seizures induced by PTZ in mice. PMID- 10834306 TI - Effects of natriuretic peptides (ANP, BNP, CNP) on catecholamine synthesis and TH mRNA levels in PC12 cells. AB - Atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP) and C-type natriuretic peptide (CNP) are present in adrenal chromaffin cells, and are co secreted with catecholamines suggesting that these natriuretic peptides (NPs) may modulate functions of chromaffin cells in an autocrine and/or paracrine manner. Therefore, we investigated the effects of NPs on tyrosine hydroxylase (TH: a rate limiting enzyme in biosynthesis of catecholamine) mRNA in rat pheochromocytoma PC12 cells. It was also determined whether the cyclic GMP/cGMP-dependent protein kinase (cGMP/PKG) pathway was involved in theses effects. Finally, we examined the effects of NPs on intracellular catecholamine content to confirm increase of catecholamine synthesis following TH mRNA induction. NPs (0.1 microM) induced significant increases of the TH mRNA (ANP= BNP> CNP). Also, the effects of NPs on TH mRNA were mimicked by 8-bromo cyclic GMP (1mM), and were blocked by KT5823 (1 microM) (inhibitor PKG) or LY83583 (1 microM) (guanylate cyclase inhibitor). Moreover, NPs were shown to induce significant increases of intracellular catecholamine contents (ANP= BNP> CNP). These findings suggest that NPs induced increases of TH mRNA through cGMP/PKG dependent mechanisms, which, in turn, resulted in stimulation of catecholamine synthesis in PC12 cells. PMID- 10834307 TI - Differential expression of amiloride-sensitive Na+ channel subunits messenger RNA in the rat uterus. AB - The expression of the amiloride-sensitive Na+ channel was investigated by reverse transcription-polymerase chain reaction (RT-PCR) in uteri from pregnant and non pregnant rats. Three subunits (alpha,beta,gamma) of this epithelial Na+ channel have been characterized in the rat. All three subunit mRNAs were present in day 16 pregnant uteri while only alpha and beta subunit mRNAs were detected in uteri from non-pregnant or day 1 postpartum rats. The level of expression of the alpha subunit was similar in day 16 pregnant and non-pregnant animals while the level of expression of the beta subunit was higher in pregnant than in non-pregnant rats. These findings show that amiloride-sensitive Na+ channels are expressed in the rat uterus and that mRNA expression levels of the alpha, beta and gamma subunits are selectively and differentially regulated during pregnancy in the rat. PMID- 10834308 TI - Prostaglandin E2 is variably induced by bacterial superantigens in bovine mononuclear cells and has a regulatory role for the T cell proliferative response. AB - Signal transduction in antigen presenting cells via MHC class II molecules induces production of prostaglandin E2 (PGE2) known to possess immunoregulatory potential. Since Staphylococcus aureus superantigens (SAgs) utilize MHC class II molecules as primary ligands, we wanted to know whether PGE2 is induced after in vitro SAg stimulation of bovine blood mononuclear cells (boMNC), and whether this arachidonic acid metabolite modulates the preferential SAg-induced proliferative response of bovine CD8+ T cells. SEB as well as SEA induced maximal amounts of PGE2 on day 2 of culture (1-2.5 x 10(-8) mol/l per 2 x 10(5) boMNC). PGE2 production could be inhibited completely by indomethacin (10(-5) mol/l) causing enhanced proliferation of boCD4+ T cells (174%) as well as of boCD8+ T cells (122%) between day 4 and 6 of the in vitro culture, however, only in a subset of the tested animals. Notably, the striking preference of proliferation of boCD8+ over boCD4+ T cells following SAg stimulation remained largely unchanged after inhibition of endogenous PGE2 synthesis or after addition of exogenous PGE2. Higher concentrations of exogenously added PGE2 (> or = 10(-8) mol/l) inhibited the proliferation reaction, mainly due to an increased death rate of both CD4+ and CD8+ blasts. In contrast, lower PGE2 concentrations between 10(-8)-10(-9) mol/l even slightly enhanced the proliferation of both T cell subsets, depending on the individual cell donor. Summing up: These data show that SAgs, indeed, can induce PGE2 production in boMNC which can enhance or reduce the proliferative response of bovine CD4+ and CD8+ T cells. PMID- 10834309 TI - Detection of CD45iota mRNA in murine Th1 but not Th2 clones. AB - CD45, a prototype of the receptor-like protein tyrosine phosphatase (PTPase) family, is one of the essential molecules in signal transduction through T cell receptors. Because at least 8 types of CD45 isoforms can potentially be produced by alternative mRNA splicing of exons 4, 5, and 6, the analyses at the transcription and protein levels of CD45 during the development and differentiation of T cells have been performed using RT-PCR and isoform-specific monoclonal antibodies, respectively. We report here that the ninth and smallest isoform of CD45, designated as CD45iota (CD45t), which is alternatively spiced from exons 4, 5, and 6 as well as exon 7, is present in the fetal thymus and splenic T cells of mice, and in murine Th1 clones, but not in Th2 clones. The expression of full-length CD45t mRNA as the functional CD45 PTPase was confirmed by RT-PCR analysis. Furthermore, the expression vector of CD45t was constructed, and its expression was detected in combination with anti-pan CD45 mAb and our newly established anti-LAR/CD45 PTPase domain mAb. These results suggested that CD45t might be an important isoform of CD45 for differentiation signaling of Th cells, and might be used as a marker to distinguish between Th1 and Th2 cells. PMID- 10834310 TI - Differentiation from thymic B cell progenitors to mature B cells in vitro. AB - The role of the thymic microenvironment in the development of murine thymic B cells has yet to be fully clarified. We therefore investigate the microenvironment that supports the development of mature thymic B cells (sIg+/B220+/CD43-B cells) from thymic B cell progenitors with immunophenotypes of sIg-/B220med/CD43+ cells. As we have previously reported, thymic B cells generated from these progenitors in the thymus are CD5+ B cells. We next study the in vitro condition that supports the differentiation of thymic B cell progenitors. Stromal cells (from the bone marrow or thymus), thymus-derived cell lines with the character of thymic nurse cells (TNCs) or thymic epithelial cells (TECs), or the bone marrow-derived cell line (MS-5) are tested for their ability to support B-lymphopoiesis from thymic B cell progenitors. Interestingly, thymic stromal cells (but neither stromal cells from the bone marrow nor stromal cell lines) support the differentiation of thymic B cell progenitors into thymic B cells in the presence of IL-7. Cortical epithelia (but not medullary epithelia, thymic macrophages or dendritic cells) are found to contribute to thymic B cell differentiation. Surface phenotype and Ig rearrangement analyses reveal that mature B cells generated in this condition are primarily CD5+ B cells, indicating that the thymic microenvironment (particularly cortical epithelia) determines the differentiation of thymic B cells. PMID- 10834311 TI - Effective priming of cytotoxic T lymphocyte precursors by subcutaneous administration of peptide antigens in liposomes accompanied by anti-CD40 and anti CTLA-4 antibodies. AB - Recently it has been shown that modulation of CD40 molecules on antigen (Ag) carrying dendritic cells (DC) can bypass T cell help, resulting in priming cytotoxic T lymphocytes (CTL) specific for the Ag. In the present study we attempted to prime peptide Ag-specific CTL by a new method in which a peptide Ag in liposome (liposomal peptide), consisting of phosphatidylserine and phosphatidylcholine (3:7), was administrated subcutaneously with anti-CD40 and/or CTLA-4 monoclonal antibodies (mAb) to mice. We found that the subcutaneous administration of the liposomal peptide with both anti-CD40 and anti-CTLA-4 mAb enhanced CTL responses comparing with those induced by the liposomal peptide alone or the liposomal peptide plus each mAb. It was shown that liposomes were critical for induction of the CTL activity. Flow cytometry analysis of a peptide bearing DC in lymph nodes (LN) and measurement of serum IL-12 indicated that anti CD40 mAb promoted migration of DC to the LN, where DC might differentiate and acquire ability of priming CTL. These findings provide a possibility that our procedure is applicable to cancer patients. PMID- 10834312 TI - Increase in CD45RO+ cells and activated eosinophils in chronic allergic conjunctivitis. AB - We assessed the infiltration of CD45RO+ cells in conjunctival biopsies of fifteen subjects affected by seasonal allergic conjunctivitis by means of immunohistochemistry. Correlations between infiltration of CD45RO+ cells and serum and mucosal indices of eosinophilic activation were investigated. The study was performed in autumn and all selected patients showed <> also in absence of sensitising pollens. Fifteen healthy subjects were used as controls. The semi-quantitative count of CD45RO+ cells in biopsy specimens demonstrated that positive cells were higher in allergic patients than in controls (p < 0.001) and EG2+ eosinophils were present only in biopsies of allergic patients. Furthermore, a statistically significant positive correlation (r = 0.73; p < 0.001) between CD45RO+ lymphocytes and EG2 positive eosinophils, was observed in the biopsies of allergic patients. Total serum IgE significantly correlated with CD45RO+ cells (r = 0.61; p < 0.02) and EG2+ eosinophils (r = 0.67; p < 0.01) in the conjunctiva. On the other hand serum ECP did not correlate with any histological and immunohistochemical parameters in the conjunctival biopsies. The present study shows that mild symptoms in SCA patients out of pollen season are associated with inflammation of the conjunctiva as shown by an increased number of CD45RO and EG2 positive cells. PMID- 10834313 TI - Inability of cyclophosphamide-induced tolerance to permit engraftment of pluripotent stem cells contained in a moderate number of syngeneic bone marrow cells. AB - We have previously described cyclophosphamide (CP)-induced tolerance that comprises an intravenous (i.v.) injection of 1 x 10(8) allogeneic spleen cells (SC) followed, 2 days later, by an intraperitoneal (i.p.) administration of 200 mg/kg of CP. By using this method, we were able to readily induce both long lasting mixed chimerism and skin allograft tolerance in most of H-2 matched combinations, but not in H-2 mismatched combinations. In the present study, we have investigated whether the treatment with SC followed by 200 mg/kg CP can permit engraftment of syngeneic pluripotent hematopoietic stem cells (HSC), and whether CP itself can permit the HSC engraftment. Recipient B6 (H-2b; Ly-5.2) mice received 1 x 10(7), 5 x 10(7) or 1 x 10(8) SC from syngeneic B6.Ly-5.1 (H 2b; Ly-5.1) mice and administered 2 day later 200 mg/kg CP. Using anti-Ly-5.1 and Ly-5.2 mAbs and a flow cytometry, the origins of lymphoid and myeloid cells injecting the recipients were observed over time. Chimerism was at least initially detectable in all groups treated with SC alone (without 200 mg/kg CP), but became undetectable by 32 weeks after the treatment with SC alone. In recipient B6 mice treated with B6.Ly-5.1 SC and 200 mg/kg CP, on the other hand, lymphoid-chimerism was detectable at 32 weeks in a transferred cell dose dependent manner, but granulocyte-chimerism indicating pluripotent HSC engraftment disappeared earlier than 32 weeks after the treatment. In order to further evaluate the effect of CP itself on HSC engraftment, recipient B6 mice were given various doses of CP (50-400 mg/kg) and were injected with T cell depleted 1 x 10(7) BMC from B6.Ly-5.1 mice. Multilineage mixed chimerism over 32 weeks was induced in only one of 11 B6 mice treated with 400 mg/kg CP followed by B6.Ly-5.1 BMC, although lymphoid chimerism was induced temporarily in recipient B6 mice treated with 200 mg/kg CP followed by B6.Ly-5.1 BMC and persistently in most of recipient B6 mice treated with 400 mg/kg CP followed by B6.Ly-5.1 BMC. PMID- 10834314 TI - Dendritic cells lose ability to present protein antigen after stimulating antigen specific T cell responses, despite upregulation of MHC class II expression. AB - Immature dendritic cells (DC) take up, process and present protein antigens; mature DC are specialized for stimulating primary T cell responses with increased expression of MHC class II and co-stimulatory molecules, but are incapable of processing and presenting soluble protein. The current study examined whether maturation of DC is triggered by T cell recognition of antigens presented by immature DC. Human DC derived from CD34+ progenitor cells by culture with granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-6 (IL 6) in serum-free medium could prime naive CD4+ T cells to keyhole limpet hemocyanin (KLH) and ovalbumin (OVA). The cultured DC retained the ability to prime T cells to native protein for at least 15 days. To test for changes in DC function after participation in an immune response, DC were co-cultured with either allogeneic or autologous CD4+ T cells. DC co-cultured with autologous T cells retained the ability to prime T cells to intact protein antigens. By contrast, DC which had previously stimulated an allogeneic T cell response lost ability to prime T cells to soluble proteins. However, such <> induced a MLR and stimulated peptide-specific primary CD4+ T cell responses. This indicated that <> did not die or lose the ability to prime, but lost the ability to process and present subsequent antigens. Following participation in T cell activation, DC increased surface expression of MHC class II, co-stimulatory molecules CD40 and B7.2, and the intercellular adhesion molecule-1 (ICAM-1). In addition, our data suggest that interferon gamma (IFN gamma) and tumor necrosis factor alpha (TNF-alpha) are involved in this T cell mediated DC maturation. PMID- 10834315 TI - Porcine MCP gene promoter directs high level expression of human DAF (CD55) in transgenic mice. AB - Porcine membrane cofactor protein (pMCP), a complement regulatory protein, is widely expressed in various tissues. Particularly, it is highly expressed on vascular endothelium. The objective of this study was to investigate whether the pMCP gene promoter can induce efficient expression of a human complement regulatory protein, decay-accelerating factor (DAF; CD55) in transgenic mice. Two fragments of the 5'-flanking region of pMCP gene (0.9 kb and 5.4 kb) connected with human DAF minigene (0.9/hDAF and 5.4/hDAF) were used to produce transgenic mice. The expression of hDAF in heart, liver, kidney, lung, pancreas, brain and testis of the transgenic mice was examined by immunohistochemical analysis. The vascular endothelia and the nerves in all organs examined were intensely stained. The staining pattern in these tissues was similar in all transgenic mice examined regardless of the length of the promoters. The surface expression levels of hDAF on peripheral red blood cells and splenocytes from a mouse carrying 5.4/hDAF hemizygously was twice the level of expression on corresponding human cells. The red blood cells and splenocytes from the transgenic mice exhibited resistance to lysis by human serum in a manner dependent upon expressed hDAF level. The hearts from the transgenic mice functioned for a significantly longer time than those from normal mice under perfusion with human serum in the Langendorff perfusion system. These results demonstrated that the pMCP gene promoter is a good candidate of the regulatory element in the transgene to produce transgenic animals for xenotransplantation. PMID- 10834316 TI - Expression of nonspecific esterase (NSE) by thyroid follicular epithelium as a marker for the target organ susceptibility to immune system attack. AB - Spontaneous autoimmune thyroiditis in obese strain (OS) chickens provides an excellent animal model for the study of Hashimoto's autoimmune thyroiditis in humans. The data presented in this paper indicate that nonspecific esterases (NSE) may play a role in or serve as a marker for the target organ susceptibility. Experiments have shown that follicular epithelial cells and interfollicular macrophages in connective tissue stain positively for NSE as early as the first day after hatching, a time at which infiltrating lymphocytes are not yet observed. We also have observed NSE positivity of follicular cells in the vicinity of mononuclear cell infiltration in all OS chickens, as well as weaker positivity in 6-month-old, avian leukosis virus free, Brown Leghorn outbred chickens, which appears in each case to correlate with infiltration of lymphocytes. In F2 hybrids between OS and healthy CB inbred chickens, the intensity of NSE staining was more variable than in OS chickens. Using specific inhibitors eserine, Na-taurocholat and p-hydroxymercuribenzoic acid, we were able to inhibit in vitro the NSE positivity of thyroid gland follicular epithelium, indicating that this staining was not an artifact. Experiments are currently in progress to clarify the relationship between the presence of NSE in follicular epithelium and the predisposition to spontaneous autoimmune thyroiditis. PMID- 10834317 TI - Increase in immune activation, vascular endothelial growth factor and erythropoietin after an ultramarathon run at moderate altitude. AB - The present study was performed to investigate the effects of exhaustive long lasting exercise at moderate altitude on the time course of serum immunomodulatory peptides, vascular endothelial growth factor (VEGF) and serum erythropoietin (EPO). Thirteen well trained runners participated at the Swiss Alpine Marathon of Davos (distance 67 km, altitude difference 2300 m). Interleukin-6 was significantly elevated in the first 2h after the run. In contrast, tumor necrosis factor-alpha and both soluble tumor necrosis factor-a receptors I and II were increased after exercise termination and showed sustained serum concentrations the following days. Neopterin, a serum marker for the activation of the cellular immune system, was increased until day two after the run. Immediately after the run VEGF was significantly elevated and further increased 2.4-fold until day five post exercise (p = 0.005). EPO was also increased after exercise but reached its maximum 2 h after the run (2-fold increase; p = 0.004) and decreased thereafter. The main findings of our study are that prolonged strenuous exercise at moderate altitude induced a significant long lasting increase in serum VEGF and EPO which was accompanied by an activation of the immune system. PMID- 10834318 TI - Enhanced tryptophan degradation in systemic lupus erythematosus. AB - In vitro and in vivo, tryptophan degradation was found to be associated with T cell functional loss and tolerance induction. In systemic lupus erythematosus (SLE) besides the Th2-type cytokine interleukin-10, Th1-type cytokines including interferon-gamma (IFN-gamma) are expressed especially during exacerbation of the disease. IFN-gamma stimulates the enzyme indoleamine (2,3)-dioxygenase (IDO) converting tryptophan to the metabolite kynurenine which in macrophages is subsequently degraded to other, partly neurotoxic compounds like quinolinic acid, and finally to nicrotinamides. We measured kynurenine and tryptophan concentrations in the sera of 55 SLE patients. In these patients, the concentrations of tryptophan (median, interquartile range: 53.9, 45.7-64.1 microM) were lower (p < 0.0001), and the kynurenine concentrations (2.45, 1.75 3.40 microM) were increased (p < 0.0005) compared to healthy blood donors (70.0, 63.8-80.6; 1.80, 1.45-2.27 microM, respectively). Also the kynurenine per tryptophan quotients (K/T), which allow to estimate IDO activity, were significantly higher in patients than in normals (0.043, 0.033-0.062 vs. 0.027, 0.021-0.030; p < 0.0001), indicating enhanced IDO-induced tryptophan degradation in SLE. There was no significant relationship between tryptophan, kynurenine and the SLEDAI, and also the correlation of K/T with SLEDAI was rather weak (rs = 0.243, p < 0.05). Higher K/T was found in patients presenting with serositis (p = 0.01), decrease of complement (c3, c4; p < 0.01) and blood count change (anemia, leucopenia, lymphopenia; p = 0.032) than in patients without such disease manifestations. The significant correlation found between K/T and neopterin (rs = 0.808, p < 0.001), a marker of immune activation, points to a role of immune activation to be responsible for tryptophan degradation in SLE patients. PMID- 10834319 TI - Clonal evolution in a primary cutaneous follicle center B cell lymphoma revealed by single cell analysis in sequential biopsies. AB - B cell neoplasias descending from germinal center cells harbor the hallmark of intraclonal diversity resulting from ongoing mutation in the variable parts of their immunoglobulin-encoding genes. To characterize a primary cutaneous follicle center B cell lymphoma in more detail, we analyzed the respective VH and VL genes in single cells mobilized from four sequential biopsies, three taken from the skin and one obtained after internal dissemination from a retrobulbar infiltrate. The lymphoma cells were found to contain V5-51/D6-12/JH5b (heavy chain) and A27/Jkappa2 (light chain) gene rearrangements detected on both the genomic and the transcriptional level. To provide an accurate mutation analysis, the specific VH gene counterpart (V5-51UK) was cloned from the patient's germline. Analyzing 226 single cells, we found: (i) complete nucleotide identity when VH and VL genes of lymphoma cells from one particular biopsy were compared among each other; (ii) intraclonal diversity due to ongoing mutation comparing the sequences obtained from sequential biopsies; (iii) both VH and VL genes to be highly mutated. Deducing from the sequence data, we propose a scenario of the clonal evolution of the B cell tumor in this patient. From the molecular-biological point of view, this primary cutaneous follicle center B cell lymphoma shows the features of a germinal center cell lymphoma. To draw this conclusion from single cell PCR data, however, a sample of sequential biopsies had to be analyzed. PMID- 10834320 TI - Maternal levels of vitamin E in normal and preeclamptic pregnancy. AB - Vitamin E, a potent antioxidant, may play a role in preventing preeclampsia. Maternal blood samples were collected between 28 and 40 weeks' gestation from women with mild preeclampsia (n=17), women with severe preeclampsia (n=16) and the control group (n=15). This control group was consisted of 15 pregnant women without hypertension episode during their pregnancy. Vitamin E levels were significantly higher in normotensive pregnant women (1.00+/-0.20 mg/dL) than in those with mild (0.56+/-0.15 mg/dL) or severe (0.37+/-0.75 mg/dL) preeclampsia (P<0.001). In preeclamptic women, when systolic blood pressure increases, maternal levels of vitamin E significantly decrease (P<0.05), also when diastolic blood pressure increases, maternal levels of vitamin E significantly decrease (P<0.05). Measurement of vitamin E concentration in plasma may be useful as a prognostic marker of the likely development of preeclampsia. PMID- 10834321 TI - Concentrative relationship between polymorphonuclear elastase and urinary trypsin inhibitor in amniotic fluid. AB - In the amniotic fluid, urinary trypsin inhibitor (UTI) seems to inhibit polymorphonuclear elastase (PMNE) activity. The PMNE and UTI concentrations in normal amniotic fluid at 16-20 and 38-40 gestational weeks were measured. The PMNE concentration increased significantly at 38-40 weeks, whereas UTI concentration decreased significantly. According to concentrative relationships between both substances, PMNE may be activated more at the full term pregnancy. Since PMNE-induced tissue injury potentially causes degradation of amniotic collagen, the present result suggests that the quotient of PMNE and UTI concentrations is a reliable index to estimate the occurrence of rupture of the membranes. PMID- 10834323 TI - Effects of pantothenic acid on postoperative adhesion formation in a rat uterine horn model. AB - This randomized blind study evaluated the effect of pantothenic acid on postoperative adhesion formation in euestrogenic and hypoestrogenic environment. We used the rat uterine horn model in a university-based laboratory setting. Thirty-six Sprague-Dawley rats were randomized into two estrogenic environments: euestrogenic and hypoestrogenic. The hypoestrogenic condition was achieved in 21 rats by either the administration of gonadotropin releasing hormone agonist or ovariectomy. The remaining 15 rats were untreated and remained in the regular estrogenic state. The left uterine horn was subjected to a lesion by serosal denudation at laparotomy. Following the uterine horn surgery, the rats within each environment were randomized into three treatment groups: saline (control), intraperitoneal pantothenic acid and intramuscular pantothenic acid. The degree of adhesions ten days following surgery was scored by an evaluated blinded to the rat's estrogenic condition and treatment. In the hypoestrogenic environment, there were no differences in the mean adhesions scores by treatment. In euestrogenic rats, the intraperitoneal pantothenic acid group had a higher mean adhesion score than intramuscular pantothenic acid, but neither treatment mean differed from that of the saline group. There was no difference in the mean adhesion scores of the saline groups by estrogenic environment. We concluded that pantothenic acid was not found to decrease adhesions formation when administered intraperitoneally or intramuscularly at these dosages. Contrary to previous reports, the hypoestrogenic condition alone was not found to be associated with decreased adhesion formation in our study. PMID- 10834322 TI - An examination of different Percoll density gradients and magnetic activated cell sorting (MACS) for the enrichment of fetal erythroblasts from maternal blood. AB - OBJECTIVE: To increase the yield of fetal erythroblasts from the blood of pregnant women by optimising and simplifying the enrichment procedure. METHODS: By using an experimental system whereby cord blood was mixed with normal adult blood, we evaluated the recovery of fetal erythroblasts using different single or double percoll density gradients with or without subsequent enrichment by Magnetic Activated Cell Sorting (MACS). RESULTS: The recovery of erythroblasts on single density gradients increased from 3%-68% of the input erythroblasts with an increase in the density of percoll from 1075 g/l-1098 g/l. Although the use of a double density gradient did eliminate most lymphocytes, it was accompanied by a significant loss of erythroblasts. The combination with MACS enrichment, however, lead to a almost 200 fold increase in the purity of erythroblasts. CONCLUSIONS: Erythroblasts are preferentially retained by higher density gradients. To enhance their purity a subsequent more specific enrichment step, such as MACS is advisable. PMID- 10834324 TI - Serial serum human chorionic gonadotropin (hCG) levels in ectopic pregnancy and first trimester miscarriage. AB - Serial serum hCG levels were measured in 50 patients with a tubal ectopic pregnancy and 50 patients with spontaneous miscarriage of an intrauterine pregnancy. Serum samples were obtained at intervals of 3-5 d and more frequently if clinically indicated. The final diagnosis was confirmed by laparoscopy and/or dilatation and curettage. Data were analyzed using a linear regression model. Initial hCG concentrations ranged from 91 to 3,050 mIU/mL. Eighty percent of ectopic pregnancies and 35% of miscarriages were associated with rising hCG concentrations and no significant differences were noted in daily increments of hCG in the two groups (210 +/- 30 mIU/mL/day for ectopic pregnancies versus 311 +/- 55 mIU/mL/d for miscarriages). Twenty percent of ectopic pregnancies and 65% of miscarriages had falling hCG concentrations and significant differences were noted in the daily decrements of hCG for EP and AB (270 +/- 52 mIU/mL/day for ectopic pregnancies versus 578 +/- 28 mIU/mL/d for miscarriages (P < or = 0.05). PMID- 10834325 TI - The endometrium in breast cancer patients on tamoxifen. AB - We restudied histologically and immunohistochemically 17 endometrial carcinomas, 2 malignant mixed tumors and 180 endometria with benign changes during or after tamoxifen therapy. The carcinomas were subtyped according to the 1994 WHO classification. Endometrial biopsies were taken only if the endometrial thickness was > 8 mm sonographically, when a polyp was seen, or for postmenopausal bleeding. About half of the endometrial specimens showed simple or cystic atrophy, 55-76% had cystic-atrophic polyps or regressive hyperplasia. Depending upon the dose of tamoxifen, 7-19% (30 mg) to 27-36% (20 mg) showed moderate glandular proliferation. 20-33% had foci of mucinous, clear cell or serous papillary metaplasia. 68-70% revealed diffuse extensive fibrosis of the endometrial stroma. None of 11 patients biopsied before starting tamoxifen therapy had advanced endometrial glandular proliferation in the second endometrial biopsy after tamoxifen treatment. None of the 19 endometrial neoplasms after tamoxifen therapy was of the endometrioid type: 11 were mucinous adenocarcinomas, 4 clear cell carcinomas, 2 serous-papillary carcinomas, one carcinosarcoma and one malignant Mullerian mixed tumor. The reasons for discrepancies between suspicious sonograms and endometrial atrophy are discussed. PMID- 10834326 TI - Oral phenoxymethylpenicillin treatment during pregnancy. Results of a population based Hungarian case-control study. AB - The objective of the study was to examine the human teratogenic potential of oral penicillin V: phenoxymethylpenicillin treatment during pregnancy in the large population-based dataset of the Hungarian Case-Control Surveillance of Congenital Abnormalities, 1980-1996. The dataset included 22,865 pregnant women who had fetuses or newborns with congenital abnormalities and 38,151 matched pregnant women who had newborn infants without any congenital abnormality (population control group). Of 22,865 case pregnant women, 173 (0.8%) had phenoxymethylpenicillin treatments, while of 38,151 population controls, 218 (0.6) were treated by this penicillin V (crude OR 1.3 with 95% CI: 1.1-1.6). This difference was explained mainly by recall bias and confounders because adjusted OR for medically documented phenoxymethylpenicillin treatments did not show difference during the second-third months of gestation, i.e. in the critical period for most major congenital abnormalities in case-matched control pairs. Thus, treatment with oral phenoxymethylpenicillin during pregnancy presents very little if any teratogenic risk to the fetus. PMID- 10834327 TI - Prenatal findings in Brachmann-de Lange syndrome. AB - We present a case of Brachmann-de Lange syndrome, in which prenatal ultrasonographic evaluation demonstrated increased nuchal translucency, early onset of intrauterine growth retardation, and limb abnormalities in the first, second, and third trimester, respectively. PMID- 10834328 TI - Delayed interval delivery in multiple gestations. AB - Two multiple pregnancies with delayed delivery after expulsion of dead fetus are presented. Case 1: A woman with a twin pregnancy and one intrauterine fetal death at 20 weeks' gestation delivered a dead fetus at 27 weeks' gestation. She delivered a healthy male infant weighing 2430 g at 33 weeks' gestation (42 d after the delivery of the first twin). Case 2: A woman with quadruplets pregnancy (2 live fetuses, one empty sac, and one fetocide at 7 weeks' gestation) got a intrauterine fetal death at 21 weeks' gestation at one fetus among 2 live fetuses and delivered a dead fetus at 24 weeks' gestation. She delivered a healthy female infant weighing 2110 g at 33 weeks' gestation (58 d after the delivery of a dead fetus). On the basis of our experience and the review of literature, delayed delivery with careful observation of fetal and maternal condition is recommended for improved survival and decreased morbidity among latter-born siblings. PMID- 10834329 TI - Benign teratoma in an 85-year-old woman. AB - BACKGROUND: In women over the age of 50, a mature cystic teratoma is reportedly likely to change into a malignant form. A mature cystic teratoma that remained in a benign form in a very old woman is a rare event. We report a case of a benign cystic teratoma, which is the oldest in Japan. CASE: An 85-year-old Japanese woman with a pelvic mass measuring 8.90x5.81 cm by ultrasonography was referred to our Department for detailed. Transvaginal ultrasonography, magnetic resonance imaging, and serum concentrations of tumor markers led to a diagnosis of a mature cystic teratoma. Histological findings postoperatively confirmed the presence of a mature cystic teratoma. CONCLUSIONS: This case report describes a mature cystic teratoma which is the second oldest case reported in the world. We reconfirmed that a mature cystic teratoma in elderly women is not frequent, but does occur. PMID- 10834330 TI - Fibroadenoma of the vulva. AB - A well-circumscribed mass was excised from the vulva of a 37-year-old woman. Histological and immunohistochemical findings showed the characteristic features of fibroadenoma of breast. The concepts about the histogenesis of this lesion were discussed. PMID- 10834331 TI - Sexual assault under benzodiazepine submission in a Paris suburb. AB - Sexual assaults under benzodiazepine submission have been described, since use of benzodiazepine enables non consensual sexual activity but rarely fully reported. An accurate evaluation of the phenomenon has seemed interesting. Files of 23 adult males and females examined at the Emergency Forensic Unit of an University Teaching Hospital near Paris were reviewed. All the victims had complained from sexual assault under drug submission, in the years 1996 and 1997. A complete examination for sexual assault was realised linked to clinical examination of drug intoxication. Every victim of rape under drug submission was sampled for urine screening (mean delay of 17.5 h after sexual assault) and blood alcohol level quantification. Urine was screened for benzodiazepines, cocaine, opiates and cannabinoids with qualitative immunochromatographic test. Traumatic lesions of sexual penetration were retrieved in 10 victims and sperm in 5. Clinical signs of benzodiazepine intoxication were retrieved in 12 out of 23 victims. Urine benzodiazepine screening was positive, over the cut-off values (300 ng/mL)when sampled less than 20 h after the facts. In 6 out of 23 victims, drugs of abuse and alcohol were associated to benzodiazepines. A reinforced attention can be brought to the rape under drug submission including the need of a proper examination and samplings shortly after the alleged facts to ascertain the diagnosis and to help the victim facing the Justice inquiry. PMID- 10834332 TI - Adenomyomatous polyp mimicking hydatidiform mole on ultrasonography. AB - The authors present a case report of a 62-year-old woman who experienced irregular genital bleeding. Although the serum hCG level was extremely low, transvaginal ultrasonography revealed vesicle pattern in the uterine cavity, suggesting trophoblastic disease. Hysterectomy was performed and histological diagnosis was adenomyomatous polyp. To our knowledge, this is the first report that the adnomyomatous polyp can demonstrate the vesicle pattern on ultrasonography. PMID- 10834333 TI - Androgen receptor gene mutation identified by PCR-SSCP and sequencing in 4 patients with complete androgen insensitivity syndrome. AB - To study the genetic defect of the human androgen receptor (hAR) gene in the complete androgen insensitivity syndrome (CAIS), we amplified each of the eight exons by PCR in genomic DNA extracted from the paraffin blocks of the resected gonads. We analyzed using SSCP, and directly sequenced the abnormally shifted bands. Mutations were found in 4 cases of CAIS. Patient 1 carried a point mutation; a G to A transition in exon 7 resulted in a change from arginine to glutamine at codon 831. Patient 2 carried a point mutation; a C to T transition in exon 7 resulted in a change from arginine to stop at codon 831. Patient 3 carried a point mutation and deletion in exon 7. A point mutation was an A to G transition that caused a glutamine to be substituted for the asparagine present at codon 819. A deletion of a G at codon 820 resulted in a frameshift and consequently in the introduction of a premature stop at codon 821. Patient 4 carried a mutation in 5' splice donor site of intron 7; a G to T transition might have caused an abnormal splicing of the exon 7. All of the mutations were found in exon 7. These mutations of hAR gene might be related to the pathogenesis of CAIS. PMID- 10834335 TI - Length and intensity of rehabilitative involvement. PMID- 10834334 TI - Secondary acute nonlymphocytic leukemia following successful chemotherapy combining cisplatin, doxorubicin, and cyclophosphamide for stage IV epithelial ovarian cancer. AB - Although chemotherapy is indispensable for the treatment of ovarian cancer, secondary acute leukemia has become increasingly important as one of the most unfavorable late effects according to widespread long-term chemotherapy. We report a patient suffering from acute nonlymphocytic leukemia (ANLL) 3 years after treatment for stage IV ovarian cancer began. PMID- 10834336 TI - Conceptual and methodological challenges in discourse assessment with TBI speakers: towards an understanding. AB - The primary objective of this paper is to review theoretical and methodological literature pertaining to the clinical evaluation of discourse abilities in speakers who have sustained traumatic brain injury (TBI). A brief history of the study of discourse impairment in this population is followed by consideration of the following issues: (1) sampling (genres selected, the physical setting in which sampling takes place, the relationship between speakers, elicitation techniques, presence of recording devices, the number of samples required, and transcription); (2) measurement; (3) the relationship between sampling and measurement; (4) other approaches to discourse assessment (self and close other report); (5) consideration of the criterion of 'normal' which clinicians should employ; (6) the relationship between discourse impairment and measures of executive function; and (7) the relationship between discourse impairment and seventy of injury. Recommendations arising from a critical review of these domains are made for both clinical practice and research. PMID- 10834337 TI - Use and need for post-acute services following paediatric head injury. AB - This paper aims to document the types of inpatient and outpatient post-acute services children receive after discharge from an acute care hospital for head injury and to better understand the extent to which children fail to receive services and the reasons for not receiving needed services. A follow-up was conducted on 95 children (aged 5-15) 1 year after they were hospitalized for head injury. Parents were interviewed by phone concerning their child's use of and need for medical, rehabilitation, and social services since the injury. Questions were also asked regarding the child's current health status and behaviour. Inpatient records were reviewed to obtain information on the characteristics of the injury. Overall use of outpatient rehabilitation and social services was low during the year following injury, ranging from 0-18% of the study sample. Although need for and use of services was positively correlated with head injury severity, it appears that unmet need was highest for children with the least severe head injuries. Finally, need for physical or occupational therapy and mental health services was unrecognized for one third of children with physical limitations and 40% of children with at least 14 identified behaviour problems. These findings underscore the need for physicians and other health care professionals to thoroughly evaluate children during follow-up visits as well as during the initial hospitalization for head injury-related deficits. Identification of functional deficits or behavioural problems should be followed up by evaluation and treatment by qualified rehabilitation professionals. PMID- 10834338 TI - Organic personality disorder after traumatic brain injury: cognitive, anatomic and psychosocial factors. A 6 month follow-up. AB - OBJECTIVE: The purpose of the study is to describe psychosocial adjustment in patients who present Organic Personality Disorder (OPD) after TBI in relation to patients with TBI without OPD. METHOD: The group included patients who were admitted as inpatients in the Neurology Service. Exclusion criteria were: previous personality disorders; previous alcohol and drugs addiction, history of head injury and other neurological diseases. For this purpose, a semi-structured interview based on the ICD-10 was applied to the patient or significant other during the 1st or 2nd week after the accident. Selected patients were evaluated with psychological and psychosocial tests and questionnaires 6 months after head injury, among them: WAIS, Benton Test, Rey Osterrieth Test, Wisconsin Cards, Psychosocial Scale and Neurobehavioural Rating Scale (NRS-27). RESULTS: No significant differences were observed in relation to demographic characteristics, type of head injury, GCS, or psychometric results. Significant differences were found in the answers to neurobehavioural and psychosocial questionnaires, showing more impairment in patients with OPD. CONCLUSIONS: The results show that, in this group, patients with OPD after TBI present more psychosocial adjustment and emotional problems than patients with TBI without OPD diagnosis. The difference found is independent of cognitive impairments. PMID- 10834339 TI - Communication abilities and work re-entry following traumatic brain injury. AB - This exploratory study was completed to determine if communication measures could discriminate employed from unemployed individuals with traumatic brain injury (TBI). Twenty adults with TBI participated, 10 employed and 10 unemployed; subjects in both groups were 1-4 years post-injury, with comparable severity of injury and type of work. Subjects were administered communication tests measuring auditory processing, effects of speaking under time pressure, production of oral language, and functional verbal reasoning ability. An aphasia test and a functional outcome measure were also administered. Results revealed that a combination of three tests, one test of functional verbal reasoning and two tests of auditory processing, correctly classified 85% of subjects as employed or unemployed. Tasks that were impairment- and disability-based appear to be more related to outcome than impairment-level tasks alone. Impairment and disability level communication tasks may provide functional and practical information, which could assist in work re-entry. PMID- 10834340 TI - Evaluation of an electronic memory aid in the neuropsychological rehabilitation of prospective memory deficits. AB - Five subjects with acquired memory impairment were trained to use a new electronic memory aid, the Voice Organizer, to manage prospective memory errors. Performance was assessed on two measures analogous to everyday memory tasks; a Message-Passing task requiring prospective recall after a delay of 9 hours, and a Domestic Task measure which involved the recall of household chores following a delay of 1-6 days. All subjects benefited from the introduction of the aid on the Message-Passing task and all but one improved their prospective recall on the Domestic Task measure. These results suggest that the Voice Organizer may be a useful aid in the neuropsychological rehabilitation of prospective memory impairment. PMID- 10834341 TI - Rehabilitation of a person with severe traumatic brain injury. AB - A case study report of a long and intensive rehabilitation programme for a young woman after she sustained a severe diffuse axonal injury in a motor vehicle accident is described in detail. The purpose of this paper is to encourage specialist brain injury rehabilitation services to offer extended rehabilitation programmes to patients, even with very severe injuries. Significant functional improvements and enhanced quality of life frequently reward the high cost and hard work involved. PMID- 10834342 TI - The man who called himself 'hockey stick': a case report including misidentification delusions. PMID- 10834343 TI - Traumatic brain injury, depression and cannabis use--assessing their effects on a cognitive performance. AB - Patient RB is presented as a demonstration of the need for a more thorough understanding of the interactions of patient variables. RB had a history of traumatic brain injury along with current mood disorder and cannabis use. It was unclear initially whether or not his cognitive impairment was a permanent result of the brain injury, or a temporary effect of his mood and/or drug use. The literature offers few guidelines or precedents for understanding such complex cases or suggesting at which level it may be most appropriate to intervene. The impact of cannabis use in this individual appeared to have a detrimental effect on his mood. Treating RB's mood disorder resulted in larger cognitive gains than would have been anticipated in the literature. Specific neuropsychological tests are identified as being particularly sensitive to the cognitive changes in mood disorders. PMID- 10834344 TI - Physical activity and oxidative stress during aging. AB - Physical activity and exercise have several beneficial effects for physical and psychological health in young and aged subjects. Exercise may reduce age-related lean body mass loss and risk for several chronic diseases including coronary artery disease, hypertension, non-insulin-dependent diabetes mellitus, anxiety, depression, functional decline, and frailty. Exercise, however, especially when performed strenuously, is associated with increased production of reactive oxygen species, able to consume endogenous antioxidants and eventually to damage biological molecules and key cellular components. Therefore, the balance between beneficial and potentially harmful effects of exercise might be of particular importance in the elderly, in which nutritional deficiencies, sedentary lifestyle, and comorbidity commonly concur to a depletion of the antioxidant reservoir of the organism and increased susceptibility to oxidative stress. The aim of this review is to summarize current experimental, clinical, and epidemiological knowledge regarding known associations and potential links between oxidative stress and physical activity/exercise during aging. Before a final recommendation can be made with respect to the possible preventive and therapeutical role of antioxidant supplementation in aged exercising people, there is a substantial need for further studies to be performed on this topic. PMID- 10834345 TI - Spatial and temporal gait variable differences between basketball, swimming and soccer players. AB - The gait variables of 10 swimmers, 10 basketball players, and 16 soccer players were compared. They were all male and right-handed. There was no statistical difference between the three groups in age, weight and height. Spatial and temporal gait variables were measured with the Bessou gait analyzer. In the swimmers group, the gait variables of the right side were not statistically different from those of the left side. The right propulsion double support duration, right cycle duration, and right late swing phase duration were respectively longer than those on the left side for the basketball players. The right propulsion double support duration, right step length, and right late swing phase duration were higher than those on the left side for the soccer players. Moreover, a discriminant analysis performed with the gait variables permitted significant differentiation between the three groups. In conclusion, both basketball and soccer players presented asymmetrical gait variables, that have never been previously reported in normal subjects, or in swimmers. These results suggest that the anticipatory postural adjustments programmed to be used just before a jump or a shoot influence the motor program of the spontaneous locomotion. These gait asymmetries could also be due to asymmetric muscle development. PMID- 10834346 TI - Swim training improves myocardial resistance to ischemia in rats. AB - As the relationship between training and ischemic heart disease is not yet unraveled, we test the hypothesis that, in a model free from environmental, behavioural, and neuro-hormonal factors, endurance training improves myocardial resistance to ischemia. As carbohydrate metabolism is relevant for myocardial resistance to ischemia, we also test whether hyperglycemia blunts the protective effect of training. Eight-week old rats were randomly assigned to four groups (n = 6-8): sedentary or trained (3-week swim program, up to 2 h/day), and normal or high-carbohydrate diet (50 g/l sucrose in drinking water). Excised hearts were perfused isovolumically (flow = 15 ml/min) with Krebs-Henseleit (2 mM free Ca++, 11 mM glucose, pH 7.38 +/- 0.02, PO2 = 670 +/- 6 mmHg, PCO2 = 43 +/- 1 mmHg, mean +/- SE), exposed to 60 min low-flow (1.5 ml/min) ischemia, and then reperfused for 30 min (15 ml/min). In normally fed rats training increased the stroke volume index (97.5 +/- 13.0 vs. 72.6 +/- 6.2 microl, P = 0.05), depressed diastolic contracture (+2.3 +/- 2.0 vs. +24.2 +/- 6.7 mmHg, P = 0.02), improved the recovery of developed pressure x heart rate (33.8 +/- 2.3 vs. 24.1 +/- 3.3 mmHg/min/1000, P = 0.05), and decreased arrhythmias (P = 0.05). In high carbohydrate-fed rats training induced myocardial hypertrophy (1.95 +/- 0.08 vs. 1.67 +/- 0.03 g, P = 0.02) and decreased arrhythmias but did not affect stroke volume, developed pressure x heart rate, and diastolic contracture. Thus endurance training improves myocardial resistance to ischemia but a high carbohydrate diet partially blunts this protection. The occurrence of an inducible alteration able to modulate myocardial tolerance to ischemia may give clues to extend our knowledge of ischemic preconditioning. PMID- 10834347 TI - Comparison of muscle oxygen consumption measured by near infrared continuous wave spectroscopy during supramaximal and intermittent pedalling exercise. AB - The two purposes of the present study were 1) to determine the oxygen consumption in working skeletal muscle from the oxygenation measured by near-infrared continuous-wave spectroscopy (NIRcws) with the arterial occlusion method during the resting condition, INT(VT), and INT(MAX) and 2) to examine whether the decline rate of oxygenation is related to maximal oxygen uptake. Eight healthy males (aged 19.8 +/- 0.4 yr, height 166.9 +/- 17.4 cm, weight 62.1 +/- 2.5 kg, and maximal oxygen uptake [VO2max] 55.9 +/- 1.9 ml/kg x min(-1)) took part in this study. The oxygenation was measured by NIRcws during the Wingate anaerobic test (WAnT) and two intermittent pedalling exercises of VT (INT(VT)) and maximal (INT(MAX)) work intensity. The decline rates of oxygenation obtained during the resting condition, INT(VT), and INT(MAX) with arterial occlusion were 0.43 +/- 0.05%/sec, 4.94 +/- 0.31%/sec, and 8.16 +/- 0.38%/sec, respectively, and that during the WAnT without arterial occlusion was 8.73 +/- 0.49%/sec. The decline rate of oxygenation during the WAnTwas significantly (p < 0.0001) related to maximal oxygen uptake (VO2max). These findings indicate that O2 is utilized from the early phase, even during a supramaximal pedalling exercise, and that the oxidative metabolic capacity may be a factor contributing to supramaximal exercises. Therefore the arterial occlusion method with NIRcws is suitable for the evaluation of the muscle O2 consumption during exercise noninvasively. PMID- 10834348 TI - The effects of physical exercise on the concentrations of ferritin and transferrin receptor in plasma of female judoists. AB - The aim of the study was to assess the effect of physical exercise on the changes in concentrations of ferritin and soluble transferrin receptor (sTfR) in plasma in course of 10 consecutive days of a training camp. Ten female elite judoists, aged 17-23 years, participated in the study. Mean concentration of ferritin for the 10 day period was 62.8 x 1.633(+/-1) microg/l, the within-subject, day-to-day variability being very high (13-75%). Ferritin concentrations were significantly correlated with the training load on the preceding day (r = 0.397; p < 0.001). Mean level of sTfR was 2.56 x 1.291(+/-1) mg/l and its within-subject variability was much lower (4-16%). Although the training loads had an effect on the intravascular hemolysis as indicated by a significant, negative correlation between load scores and haptoglobin concentration (r = -0.282; p < 0.01), the latter was not correlated significantly with sTfR levels. It was concluded that the soluble transferrin receptor is a more stable indicator of iron status under high training loads since, unlike ferritin, it does not respond to the workload on the preceding day. Moreover, the intravascular hemolysis observed in athletes does not affect the sTfR levels in plasma. PMID- 10834349 TI - Relationship among oxygenation, myoelectric activity, and lactic acid accumulation in vastus lateralis muscle during exercise with constant work rate. AB - The purpose of this study was to determine whether oxygenation in localized working muscle depended on the muscle activity and on the lactic acidosis level. Seven healthy male subjects underwent the five 6-min cycling exercises with work rates of 50 watts (25.0 +/- 5.0% VO2max), 100 watts (36.6 +/- 6.2% VO2max), 150 watts (50.6 +/- 7.7% VO2max), 200 watts (67.8 + 6.9% VO2max), and 250 watts (82.9 +/- 7.5% VO2max) while gas exchange parameters and blood lactate concentration (BL) were measured. We also measured oxygenated hemoglobin and myoglobin concentration (oxy-Hb/Mb) with continuous-wave near infrared spectroscopy (NIR) and surface myoelectric activity with surface electrodes (EMG). The NIR probe and electrodes were positioned on the vastus lateralis muscle of the right leg. The relative change in oxy-Hb/Mb was estimated by regarding oxy-Hb/Mb in the resting condition as 100% and that obtained during thigh occlusion as 0%. The mean values of oxy-Hb/Mb and integrated EMG (iEMG) were determined from 5'30" to 6'00" at each work rate. The percentage of oxy-Hb/Mb was sustained at the first two work rates corresponding to 25.0 +/- 5.0 and 36.6 +/- 6.2% VO2max and decreased slightly at 150 watts corresponding to 50.6 +/- 7.7% VO2max, which was followed by a linear decrease at 200 and 250 watts corresponding to 67.8 +/- 6.9 and 82.9 +/- 7.5% VO2max. The iEMG, however, was increased slowly at 25.0 +/- 5.0 to 50.6 +/- 7.7% VO2max, and a rapid increment of the iEMG occurred at 67.8 +/- 6.9 and 82.9 +/- 7.5% VO2max. BL was sustained at 25.0 +/- 5.0 to 50.6 +/- 7.7% VO2max and increased linearly at 67.8 +/- 6.9 and 82.9 +/- 7.5% VO2max. There was a significant negative correlation for each subject between the percentage of oxy Hb/Mb and iEMG (r = -0.947 to -0.993), between the percentage of oxy-Hb/Mb and BL (r = -0.890 to -0.982), and between the percentage of oxy-Hb/Mb and VO2 (r = 0.929 to -0.994) These results indicated that oxygenated hemoglobin/myoglobin concentration measured with NIR reflected the muscle activity and the lactic acidosis. PMID- 10834350 TI - Metabolic response during sport rock climbing and the effects of active versus passive recovery. AB - The objectives of this study were to 1) continuously assess oxygen uptake during and after difficult sport rock climbing and 2) to evaluate the effects of active versus passive recovery on post-climbing blood lactate and hand grip strength. Fifteen expert rock climbers attempted to climb (i.e., red point lead) a 20 m difficult route (5.12 b, YDS scale) set on an indoor climbing wall. Subjects were assigned to either active recovery (AR; n = 8), consisting of recumbent cycling at 25 Watts, or passive recovery (PR; n = 7). Expired air was analyzed during climbing and through a 10-minute recovery period by a lightweight battery-powered open circuit system. Oxygen uptake (VO2) and heart rate (HR) were measured continuously and averaged over 20-second intervals. These data were expressed as averages over the entire climb (VO2avg and HRavg) and as peak values. An estimated resting VO2 of 250 ml x min(-1) was subtracted from the interval VO2 values to provide net VO2 data which were subsequently converted to absolute VO2 values in liters for climbing (C - VO2net) and recovery (R - VO2net). Total net VO2 was calculated as the sum of C - VO2net plus R - VO2net. Blood samples were obtained via fingerprick at pre-climb and at 1-, 10-, 20-, and 30-minutes post climb and analyzed for whole blood lactate. Handgrip strength was measured via dynamometry at pre-climb and at 1-, 10-, 20-, and 30-minutes post-climb. Mean climbing time was 2.57 +/- 0.41 min. During climbing, VO2avg and HRavg means were 1660 +/- 340 ml x min(-1) and 148 +/- 16 b x min(-1) respectively with mean peaks of 2147 +/- 413 ml x min(-1) and 162 +/- 17 b x min(-1). Relative VO2avg was 24.7 +/- 4.3 ml x kg(-1) x min(-1) with a mean peak value of 31.9 +/- 5.3 ml x kg(-1) x min(-1). Mean values for C - VO2net and R - VO2net were 4.009 +/- 0.929 L and 2.809 +/- 0.518 L respectively for the PR group with mean total net VO2 at 6.818 +/- 1.291 L. For the AR group mean values for C - VO2net and R - VO2net were 4.216 +/- 1.174 L and 7.691 +/- 3.154 L respectively with a mean total net VO2 of 11.906 +/- 4.172 L. There was no difference between the groups for C - VO2net, however R - VO2net and total net VO2 were significantly different (p < 0.05) between PR and AR. Blood lactate increased significantly with climbing in both AR and PR groups. Lactate remained elevated in the PR group until 30 minutes post climb, but had returned to pre-climb level by 20 minutes in the AR group. Handgrip strength was significantly decreased at 1-minute post-climb for the AR group, but was not significantly changed for the PR group. Although climbers may be able to attain a plateau in VO2, the observed accumulation of lactate in the blood combined with the elevated recovery VO2 indicate a higher overall energy demand than indicated via the recorded VO2 during climbing. Low intensity active recovery appears to significantly reduce accumulated blood lactate within 20 minutes following difficult climbing, however further research is required to establish whether this strategy is advantageous for subsequent climbing performance. PMID- 10834351 TI - Activation of blood platelets in response to maximal isometric exercise of the dominant arm. AB - Isometric exercise is a popular form of physical activity for many people. Only few studies exist on the effects of this type of exercise on the hemostatic system. Eleven male healthy subjects (21-42 years) of varying fitness levels were investigated before, immediately after and 10 min after strenuous isometric exercise of the dominant arm. Blood samples were drawn by repetitive puncture from both the exercising and the contralateral arm. The following variables were studied: Prothrombin time and partial thromboplastin time as group tests for the plasmatic coagulation system; platelet count as well as p-selectin expression for the platelet system; tissue plasminogen activator (t-PA) activity and antigen for the fibrinolytic system. The partial thromboplastin time was shortened immediately after maximal isometric exercise of the dominant arm, the prothrombin time remained unchanged. No change was found in the platelet count, but a marked p-selectin expression was observed immediately after maximal isometric exercise of the dominant arm (p < 0.05) and even in the resting contralateral arm. Values returned to baseline after 10 min. There was a slight increase of t-PA antigen concentration and white blood cell count at maximal isometric contraction which did not occur in the resting arm, although changes over the 3 time points were significant in both arms. Maximal isometric exercise leads to platelet activation in both arms, a slight aPTT decrease and t-PA antigen increase in local blood stream. As compensatory fibrinolytic changes do not occur, it is an open question whether isometric exercise increases the potential risk of thromboembolism. PMID- 10834352 TI - The validity of power output recorded during exercise performance tests using a Kingcycle air-braked cycle ergometer when compared with an SRM powermeter. AB - This study assessed the validity of power output recorded using an air-braked cycle ergometer (Kingcycle) when compared with a power measuring crankset (SRM). For part one of the study thirteen physically active subjects completed a continuous incremental exercise test (OBLA), for part two of the study twelve trained cyclists completed two tests; a maximal aerobic power test (MAP) and a 16.1 km time-trial (16.1 km TT). The following were compared; the peak power output (PPO) recorded for 1 min during MAP, the average power output for the duration of the time-trial and power output recorded during each stage of OBLA. For all tests, power output recorded using Kingcycle was significantly higher than SRM (P < 0.001). Ratio limits of agreement between SRM and Kingcycle for OBLA showed a bias (P < 0.00) of 0.90 (95%CI = 0.90-0.91) with a random error of X or / 1.07, and for PPO and 16.1 km TT ratio limits of agreement were 0.90 (95%CI = 0.88-0.92) X or / 1.07 and 0.92 (95% CI = 0.90-0.94) X or / 1.07, respectively. These data revealed that the Kingcycle ergometry system did not provide a valid measure of power output when compared with SRM. PMID- 10834353 TI - Comparison of physiological responses to open water kayaking and kayak ergometry. AB - This study compared the physiological responses of simulated kayaking on a K1 ERGO kayak ergometer with open water paddling. Nine well-trained male kayakers (VO2peak 4.27 +/- 0.58 L x min(-1), age 24 +/- 4 yr, mass 77.3 +/- 6.4 kg, height 179.5 +/- 5.3 cm; [mean +/- SD]) performed two 4 min exercise bouts on open water (OW) and on an air braked kayak ergometer (Erg). During exercise, expired air and heart rate (HR) were continuously measured. The distance covered during OW (992 +/- 47.1 m) was highly correlated (r2 = 0.86) with the total work performed in Erg (47.64 +/- 7.67 kJ). There were no differences between trials for oxygen uptake, carbon dioxide production or estimated carbohydrate oxidation. However, during OW, minute ventilation was significantly higher at 60 and 90 s (104.2 +/- 16.4 vs. 92.6 +/- 20.4 L x min(-1) and 120.5 +/- 15.8 vs. 111.7 +/- 17.6 L x min( 1) for 60 and 90 s, respectively, p < 0.05), and HR was higher in OW during the first minute (120 +/- 20 vs. 104 +/- 19 beats x min(-1), 164 +/- 8 vs. 147 +/- 18 beats x min(-1) and 178 +/- 6 vs. 170 +/- 7 beats x min(-1) for 0, 30, and 60 s, respectively, p < 0.05). There were no differences in peak VO2 between OW and Erg (4.10 +/- 0.49 vs. 4.09 +/- 0.53 L x min(-1), respectively) nor in post-exercise blood (lactate) (6.43 +/- 1.47 vs. 6.59 +/- 0.99 mmol x L(-1), respectively). We conclude that the K1 ERGO accurately simulates the physiological demands of short term, high-intensity kayaking. PMID- 10834354 TI - Energy cost and energy sources of ball routine in rhythmic gymnasts. AB - The energy cost and the different energy sources of competitive rhythmic ball routines were assessed in nine young elite rhythmic gymnasts (13-16 years of age). The overall energy requirement of ball exercise (VO2eq) was obtained by adding the amount of VO2 during exercise above resting (VO2ex) to the VO2 up to the fast component of recovery (VO2al) and to the energy equivalent of peak blood lactate accumulation (VO2la-) of recovery. The lactate (La) curve up to 25 minutes of recovery always showed La peaks at 3 min after exercise (4.0 +/- 0.4 mmol/l) and values still above rest at 25 min (2.3 +/- 0.5 mmol/l). Although ball routines were short in duration (90 s), the metabolic power requirement was 1.1 times higher than the subject's VO2max. The energy cost (VO2eq) amounted to 81 +/ 5 ml x kg(-1). Higher fractions of VO2eq were VO2ex 49% and VO2al 42%, while the remaining 9% was due to VO2la-. The aerobic source resulted in the most utilized system due to the subjects' high VO2max (52.7 ml x kg(-1) x min(-1)) and anaerobic threshold (LT = 84.4%). The HR and the peak lactate values of ball routine were similar to the values recorded at LT step during treadmill test. Although the HR and VO2 values were significantly correlated (during pre exercise, exercise, and fast recovery), the exercise intensity derived from HR Kanvonen method during ball routine did not correspond to the % of VO2max. PMID- 10834355 TI - Peripubertal perturbations in elite gymnasts caused by sport specific training regimes and inadequate nutritional intake. AB - Low body fat masses of elite female gymnasts are favoured for the current aesthetic appeal required for complex movements performed by the gymnasts. Optimal nutritional intake relative to physical training regimes is essential for pubertal development. Here we evaluate how high intensity training in combination with nutritional intake affects pubertal development. Twenty-two female (13.6 +/- 1.0 years) and 18 male (12.4 +/- 1.6 years) elite gymnasts from national cadres were enlisted in this study. Skeletal maturation and hormonal levels of the hypophyseal, gonadal, and adrenal axes were estimated. Prepubertal and pubertal stages were determined, and body composition was measured using two indirect methods. Whereas female gymnasts showed bone retardation (1.7 years), reduced height potential, minimal fat mass (4.3 +/- 1.3 kg), no significant increase in pubertal oestradiol levels (17.6 +/- 4.2 pg/ml vs. 23.9 +/- 13.4 pg/ml), and delayed menarche (2.3 years), male gymnasts displayed virtually unaltered pubertal development due to different training regimes. Nutritional intake was insufficient in all gymnasts although to a lesser extent for male gymnasts. Intensive physical training of elite female gymnasts combined with inadequate nutritional intake can alter the normal pattern of pubertal development. In female gymnasts the onset of menarche can be influenced by keeping the amount of fat mass low. There is a peripubertal change favouring fat mass over muscle mass in females while there is a net gain of muscle mass during pubertal development in males. PMID- 10834356 TI - Cancer incidence among Finnish world class male athletes. AB - A cohort of 2269 male world class athletes from the years 1920-65 was followed up through the Finnish Cancer Registry for cancer incidence during 1967-95. There were 331 cases of cancer while the expected number based on the national cancer incidence rates was 404; the standardized incidence ratio (SIR) was 0.82 and its 95% confidence interval (CI) 0.73-0.91. There was a decreased risk of cancers of the lung (SIR 0.35, 95% CI 0.25-0.48), especially among endurance athletes (0.12, 0.01-0.41). This fits well with the low prevalence of smoking among the athletes. The SIR for kidney cancer was also significantly low (0.43, 0.16-0.93), possibly partly due to the better diet (more vegetables, less fat). Incidence of cancers not related to smoking was not reduced. None of the cancer sites showed a significant excess in the whole cohort but hurdlers had a significantly high relative risk of sarcomas of the bone and soft tissue which according to suggested causal mechanisms might be related e.g. to injuries during their active sport period. This as well as other observations without sound a priori hypotheses needs to be confirmed in more detailed analyses and in other data sets before final conclusions. PMID- 10834357 TI - A longitudinal study of injuries to athletes with disabilities. AB - This study investigated the number and type of injuries that occurred to athletes with disabilities from 1990-1996. The subjects were U.S. athletes who participated in the 1990 World Games and Championships, 1991 U.S. Paralympic Trials, 1992 Paralympic Games, the 1994 World Athletics Championship, and the 1996 Paralympic Games. A reportable injury was defined as an injury/illness that was evaluated by the U.S. Medical Staff during these competitions. Illnesses (29.8%) were the most commonly reported problem followed by muscular strains (22.1%). The most commonly injured body locations were the thorax/spine at 13.3%, the shoulder complex at 12.8%, the lower leg/ankle and toes at 12.0%. The majority of these injuries were musculoskeletal to include strains at 22.1%, sprains at 5.8%, contusion at 5.6%, and abrasions at 5.1%. An area of concern was the high number of strains to the thorax and spine when compared to other body locations and injury types. Attention should be paid to the musculature of the thorax/spine, shoulder and hip/thigh to help reduce the number of the injuries in this region. PMID- 10834358 TI - Increased premature mortality of competitive powerlifters suspected to have used anabolic agents. AB - Misuse of supraphysiological doses of anabolic steroids is claimed to have serious side effects. The aim of the study was to determine the mortality, and the cause of premature deaths among a group of subjects who are strongly suspected to have used anabolic steroids for a non-medical purpose over several years. The mortality of 62 male powerlifters placed 1st-5th in weight series 82.5 125 kg in Finnish championships during 1977-1982 was compared with the mortality of population controls. The mortality during the 12-year follow-up was 12.9% for the powerlifters compared to 3.1% in the control population. By 1993 eight of 62 powerlifters and 34 of 1094 population controls had died, thus the risk of death among the powerlifters was 4.6 times higher (95% CI 2.04-10.45; p = 0.0002). The causes of premature death among the powerlifters were suicide (3), acute myocardial infarction (3), hepatic coma (1) and non-Hodgkin's lymphoma (1). These findings add to the growing amount of evidence of an association between anabolic steroid abuse and premature death, and support the view that measures to decrease AAS misuse among both competitive and amateur athletes are justified. PMID- 10834359 TI - Hemoglobin and packed-cell volume in endurance athletes prior to rhEPO. PMID- 10834360 TI - Comment on "A chemical aqueous phase radical mechanism for tropospheric chemistry" by H. Herrmann et al. PMID- 10834361 TI - Arsenic uptake in orchard trees: implications for dendroanalysis. AB - The distribution of arsenic in the stems of fruit trees grown in soils exposed to arsenical pesticides has been studied using neutron activation analysis. The results show arsenic to be confined mainly to heartwood near the pith and active xylem tissue in the most recent annual growth rings. These results suggest that dendroanalysis for arsenic in fruit trees will not yield a reliable chronology of arsenic exposure. PMID- 10834362 TI - The study of fine and coarse particles, and metallic elements for the daytime and night-time in a suburban area of central Taiwan, Taichung. AB - Daily average concentrations of fine and coarse particulates, and TSP samples have been measured simultaneously at daytime and night-time periods by using Universal and PS-1 sampler in a suburban area of central Taiwan from June to August 1998. The samples were analyzed by atomic absorption spectrometry to determine the fine and coarse particulate concentrations of metallic elements (Ca, Fe, Mn, Pb, Cu, Zn and Cr). The concentration of PM2.5 and TSP showed a decreased trend for the daytime period. The fine particle concentrations were about two times as that of coarse particulate concentrations. The averaged fine particulate concentrations at daytime are higher than at night-time. Ca and Fe were mostly in the coarse particulate mode. The correlation coefficients were 0.63 and 0.69 for elements Ca and Fe in the coarse particle mode for day and night periods. Pb showed a similar distribution ratio with Mn for the fine to coarse particle ratios at both day and night period. Pb and Mn are highly correlated for the day (R = 0.78) and night period (R = 0.61) at particle size <2.5 microm. Cu and Zn were mainly in fine particles at both day and night period. Fe and Ca consist of the major parts of all the elements. Elemental Mn is the lowest among the rest of the heavy metals. PMID- 10834363 TI - An automated test for measuring polymer biodegradation. AB - The biodegradability of polymer materials as evaluated by the modified Sturm test is labor-intensive, cumbersome and costly and also tends to cumulate errors. An automated system for the measurement of carbon dioxide would overcome many of these disadvantages. We describe here a method in which CO2 was determined by IR spectroscopy. We compared the results with those from trapping CO2 in a solution of barium hydroxide (Ba(OH)2) followed by manual titration. The automated system was more reproducible, less costly and more compact. The automated system could also be employed to measure the biodegradability of other substances such as oils and detergents. PMID- 10834364 TI - Soil organic matter mobilization by root exudates. AB - In order to study the different soil organic matter mobilisation by agrarian (Zea mais: cultivars Paolo and Sandek) and forest (Picea abies Karst. and Pinus sylvestris L.) root exudates, three different soils (Dystric Spodic Cambisol--S1, Haplic Luvisol--S2 and Calcaric Cambisol--S3) have been considered. Treating the soils with water (control) or plant root exudates, soil organic matter extracts were obtained. The extracts were characterised by hormone-like activities and gas chromatographic/mass spectrometric (GC/MS) measurements. Water extract and plant root exudates exhibited no hormone-like activity, while the other soil-extracts were endowed with a different hormone-like behaviour. GC/MS data indicated that in the acid soils (S1) Sandek and Picea abies exudates showed a greater ability in extracting organic acid isomers (Cl4COOH, Cl5COOH and Cl7COOH), while in neutral soils (S3) all the exudates were active in separating organic acids. In intermediate conditions (S2), Picea abies and Pinus sylvestris exudates liberated C15COOH isomers, Paolo C11COOH isomers, while Sandek was not effective. The different role of plant root exudates in mobilising bio-molecules from the bulk of the soil is proposed. PMID- 10834365 TI - Predictions of binary sorption isotherms for the sorption of heavy metals by pine bark using single isotherm data. AB - The adsorption of three heavy metal ions by pine bark was studied. The study was divided into two parts; single component adsorption of the metals Cu2+, Cd2+ and Ni2+ and bisolute adsorption of the three binary systems Cu2+-Cd2+, Cu2+-Ni2+ and Cd2+-Ni2+. Extended Langmuir model, extended Freundlich model. Sips model and ideal adsorption solution theory (IAST) models were used to predict the equilibrium uptake for Cu2+, Cd2+ and Ni2+ in the binary diluted solutions using the single adsorption constants. The experimental data of single isotherm adsorption process were found to follow Langmuir isotherm model with less accuracy than Freundlich and Sips models. Whereas, the predictions of bisolute adsorption isotherms of the mentioned three systems, Cu2+-Cd2+, Cu2+-Ni2+ and Cd2+-Ni2+, showed good agreement with experimental data when using Extended Langmuir, Extended-Freundlich and IAST. However, the only good fit of the Sips model was with the Cu2+-Cd2+ system. PMID- 10834366 TI - Kinetics of heterogeneous ozone reactions. AB - Earlier results on ozone destruction on solid surfaces gave apparent first order kinetics. Estimating the reaction kinetics from our data on ozone destruction on various powders (silica-gel, alumina, wood ash, coal ash, Saharan sand, calcite), we found that only calcite and wood ash exhibited such a behaviour. Removal of ozone by other powders used showed two straight lines in ln c-t plot with two different half-lives, t'(1/2) < t''(1/2). Comparing the kinetic constants for ozone removal on silica-gel and that of ozone reactions with polynuclear aromatic hydrocarbons (PAHs) adsorbed in submonolayer coverage on the same powder, the first reaction seems to be more likely in the case of pyrene and particularly fluoranthene. Enhanced ozone destruction on airborne aerosols could be an additional reason for fluoranthene stability in the real atmosphere. PMID- 10834367 TI - Aquatic fate assessment of the polycyclic musk fragrance HHCB. Scenario and variability analysis in accordance with the EU risk assessment guidelines. AB - By means of the environmental fate and distribution models laid down in the Technical Guidance Documents (TGD) and implemented in the European Union System for the Evaluation of Substances (EUSES) environmental concentrations of the polycyclic musk fragrance HHCB (1,3,4,6,7,8-hexahydro-4,6,6,7,8,8-hexamethyl cyclopenta-[g]-2- benzopyrane; trade name: e.g. Galaxolide) were calculated for the aquatic environment under consideration of various scenarios. The results were then compared to monitoring data from the region of North Rhine-Westphalia (River Ruhr). An uncertainty analysis was carried out to determine sensitive parameters, to integrate environmental variability and to confirm the model's calculations. The standard scenario of EUSES overestimates the measured concentrations, which confirms the conservative nature of the calculations. The regional-specific scenarios lead to lower deviations from the measured values than the standard scenario. Deviations range from one to two orders of magnitude in the effluent of sewage treatment plants; they amount to one order of magnitude for surface water concentrations on a local scale and conform to monitoring data on a regional scale. The use of measured bioconcentration factors for fish instead of estimated ones reduces deviations remarkably. The investigation reveals that unrealistic worst-case calculations of HHCB can at best be ameliorated by the application of more realistic emission rates and measured bioconcentration factors. The use of regional-specific parameters also diminishes the deviations of the calculations from the measured concentrations. PMID- 10834368 TI - Mass balance model of source apportionment, transport and fate of PAHs in Lac Saint Louis, Quebec. AB - A mass balance model has been developed and calibrated to describe the sources, transport and fate of seven polycyclic aromatic hydrocarbons (PAHs; anthracene, benzo(a)pyrene, benzo(b)fluoranthene, chrysene, fluoranthene, phenanthrene, and pyrene) in the water and sediments of, and atmosphere over Lac Saint Louis, Quebec. The model uses specified input rates from background advective flows and emissions from the Alcan aluminum smelting facility at Beauharnois to deduce atmospheric concentrations and rates of wet and dry deposition to the three segment lake. Concentrations in water and sediment as well as relevant mass fluxes and residence times are computed and compared satisfactorily with monitoring data for five of the seven PAHs. Underestimation of concentrations for anthracene and phenanthrene is attributed to unquantified additional sources. The sources of the PAH burden in the lake are apportioned, and the implications of these results are discussed including likely response times to changes in loadings. It is suggested that this mass balance approach is more widely applicable to situations in which water bodies are impacted by a variety of contaminant sources. PMID- 10834369 TI - Application of a purge and trap TDS-GC/MS procedure for the determination of emissions from flame retarded polymers. AB - Emissions of volatile organic compounds (VOCs) from different thermoplastic polymers used in electrotechnical applications were investigated using a purge and trap procedure that involved adsorption on Tenax GR. Results were compared to those for an operating TV set monitored in a test chamber. The analyses were in both cases carried out using thermodesorption gas chromatography with mass spectrometric detection (TDS-GC/MS). Substances identified were monomers, volatile additives, or related compounds. Special attention was given to the detection of halogenated compounds. Their origin was studied using reference samples and synthetic standards. PMID- 10834370 TI - Structural study on inclusion complexes of cyclodextrins with organophosphorus pesticides by use of rotational strength analysis method. AB - For inclusion complexes of cyclodextrins with parathinons and paraoxon, inclusion depths were estimated with the rotational strength analysis method. The inclusion depths of pesticides were sensitive to the difference in the hydrophobic properties of the phosphate and monothiophosphate groups and were in close correlations with the substrate-specificity for the cyclodextrin-catalyzed hydrolysis of pesticides. The inclusion depths were also characteristically affected by surfactant material having high solubilization functions to aquatic environmental chemicals. PMID- 10834371 TI - Anaerobic degradation and toxicity of commercial cationic surfactants in anaerobic screening tests. AB - Anaerobic biodegradability and toxicity on anaerobic bacteria of di(hydrogenated tallow) dimethyl ammonium chloride (DHTDMAC) and two esterquats have been investigated. A batch test system containing municipal digester solids as a source of anaerobic bacteria, based on the method proposed by the ECETOC, has been applied. To evaluate the potential toxicity of such surfactants on anaerobic sludge, a co-substrate, an easily biodegradable compound in anaerobic conditions, has been added to the samples to test and the effects on biogas production have been determined. For the esterquats studied high biodegradation levels were obtained and no toxic effects on anaerobic bacteria were observed even at the highest concentrations tested, 100 and 200 mg C/l, respectively. On the contrary, DHTDMAC was not degradated at the same test conditions. However, no inhibitory effects on the biogas production were detected for this surfactant at concentrations <100 mg C/l. PMID- 10834372 TI - Sorption of non-ionic surfactants to soil: the role of soil mineral composition. AB - In this study, the importance of individual soil mineral constituents on non ionic surfactant sorption is evaluated by employing scanning electron microscopy (SEM) equipped with energy-dispersive X-ray analysis (EDAX) to observe microchemical surface composition of model soil colloids and relate surface microchemical composition to the sorption behavior of a non-ionic surfactant, Polyethylene Glyco Mono-p-nonylphenyl Ether (A9PE10), on selected model soil colloids. This information is critical to understanding the fate and transport of this class of contaminants in subsurface. Results obtained from this study indicate that a correlation between the atomic ratio of Si:(Al + Fe) on soil mineral surface and A9PE10 sorption capacity exist, where those soil with larger value of Si:(Al + Fe) ratio exhibit higher sorption capacity. This allows us to predict that the non-ionic surfactant sorption capacity for soil decrease with progressively increasing states of soil maturity. PMID- 10834373 TI - Soil-to-plant and plant-to-cow's milk transfer of radiocaesium in alpine pastures: significance of seasonal variability. AB - Because our present knowledge on the environmental behaviour of fallout radiocaesium in semi-natural environments is rather limited, the transfer of this radionuclide and of natural 40K, from soil-to-plant as well as from plant-to cow's milk was investigated for a typical alpine pasture (site P). For comparison, a nearby alpine pasture (site K) not used for cattle grazing was also studied. Small seasonal effects were found for 137Cs in the plants, but they were different for the two pastures. Due to the presence of a large variety of different plant species on the pastures and soil adhesion on the vegetation from trampling cattle, the scattering of the data was very large, and the seasonal effects were observable only because of the large number of samples (N approximately 100) collected. The aggregated soil-to-plant transfer factor of 137Cs was for site P, on average, 0.002 +/- 0.001 m2 kg(-1). The plant-to-milk transfer coefficient was, on average, 0.02 day l(-1). The 137Cs concentration in the milk of the cows varied within the grazing period only between 1.4 and 2.9 Bq l(-1), with a significant maximum in the beginning of August. As a result of soil adhesion due to cattle trampling, significantly higher ash- and 137Cs contents of the plants were observed at site P as compared to site K. Possible consequences of the above observations with respect to a representative sampling design of vegetation and milk are discussed. PMID- 10834374 TI - Enantiomer fractions instead of enantiomer ratios. AB - The use of enantiomer ratios (ERs) to indicate the relative amounts of a pair of enantiomers in a sample has some disadvantages. Enantiomer fractions (EFs) are proposed as an alternative expression to eliminate the difficulties. PMID- 10834375 TI - Metal biomonitoring with mosses in the surroundings of an oil-fired power plant in Italy. AB - Levels of 12 trace elements were measured in samples of the bryophyte Hypnum cupressiforme Hedw. and in soil collected in the surroundings of an oil-fired power plant in Northern Italy. Metal bioaccumulation in moss was estimated after soil correction in order to obtain deposition patterns and individuate potentially toxic metals emitted from the plant. V and Ni, occurring together in fuel oil, showed highest bioaccumulation values near the stacks. Mean contamination of the study area for these elements is 5.5 (V) and 3.3 (Ni) times the background levels of the reference site. Other elements showed only limited alterations of bioaccumulation values, in relation to agricultural and industrial activity in the study area. PMID- 10834376 TI - Methoxyphenols from burning of Scandinavian forest plant materials. AB - Semivolatile compounds in smoke from gram-scale incomplete burning of plant materials were assessed by gas chromatography and mass spectrometry. Gas syringe sampling was shown to be adequate by comparison with adsorbent sampling. Methoxyphenols as well as 1,6-anhydroglucose were released in amounts as large as 10 mg kg(-1) of dry biomass at 90% combustion efficiency. Wood, twigs, bark and needles from the conifers Norway spruce and Scots pine emitted 12 reported 2 methoxyphenols in similar proportions. Grass, heather and birchwood released the same 2-methoxyphenols but also the corresponding 2,6-dimethoxyphenols which are characteristic of angiosperms. The methoxyphenols are formed from lignin and differ in structure by the group in para position relative to the phenolic OH group. Prominent phenols were those with trans-1-propenyl and ethenyl groups in that position. Vanillin, 4-hydroxy-3-methoxybenzaldehyde, was a prominent carbonyl compound from the conifer materials. PMID- 10834377 TI - Degradation of silicone polymer in a field soil under natural conditions. AB - Silicone polymers (PDMS = polydimethylsiloxane) are used in numerous consumer and industrial products. Our previous work showed that they will degrade in soil under laboratory conditions. This paper investigates PDMS degradation in the field. Four soil plots (each 2.44 m x 2.44 m) in Michigan were sprayed in May, 1997, with aqueous emulsion to achieve nominal soil PDMS concentrations of 0 (control), 215 (low), 430 (medium), and 860 (high) microg/g. Over the following summer, soil cores (0-5 and 5-10 cm) were collected every two weeks and analyzed for decrease in-total soil PDMS, and decrease in molecular weight of remaining PDMS. PDMS concentrations decreased 50% in 4.5, 5.3, and 9.6 weeks for the low, medium, and high treatments, respectively. Degradation rates were 0.26 (low), 0.44 (medium), and 0.44 (high) g PDMS/m2 day, indicating that degradation capacity of the soil was exceeded by the High treatment. Dimethylsilanediol (DMSD), the main degradation product, was detected in most samples at <5% of original PDMS. This is consistent with laboratory data showing biodegradation and volatilization of DMSD. Deeper sampling (to 20 cm) found only trace amounts of DMSD, and minor downward movement of the polymer. Respraying and subsequent analysis of one plot with a medium treatment in late August showed slow PDMS degradation during the cool, wet fall, followed by a 40% decrease over winter and extensive degradation during the summer of 1998. The study thus shows that PDMS will degrade under field conditions as predicted from laboratory experiments. PMID- 10834378 TI - Bisphenol A: emissions from point sources. AB - Bisphenol A is widely used for the production of epoxy resins and polycarbonate plastics. Special in vitro test systems and animal experiments showed a weak estrogenic activity for Bisphenol A. Based on in vitro receptor interaction studies, the activity was estimated to be 2 x 10(-3) lower than that of estradiol. Especially aquatic wildlife could be endangered by waste water discharges. To manage possible risks arising from Bisphenol A contamination, the major fluxes need to be obtained and the contributors to the contamination of municipal treatment plants need to be determined. In this study, industrial emitters and communal waste waters were monitored simultaneously. Mixed samples were taken over periods of one week at nine sample sites. The results showed that the concentrations and fluxes were variable. The paper industry was the major Bisphenol A contributor to the influent of the waste water treatment plant. All the other fluxes measured, including two household areas, were considerably low. About 90% of the total load was removed during the waste water treatment. PMID- 10834379 TI - 4-Nonylphenols and 4-tert-octylphenol in water and fish from rivers flowing into Lake Biwa. AB - Surveys of 4-nonylphenols (NOs) and 4-tert-octylphenol (OC) were performed for water and fish samples obtained from eight rivers flowing into Lake Biwa once every two months from April 1998 to March 1999. For water samples, NOs were detected all the year round (0.11-3.08 ng ml(-1)) at high frequency (48/48) in the eight rivers. OC was detected at lower concentrations (ND approximately 0.09 ng ml(-1)) and at lower frequency (23/48). The concentrations of NOs in the river water always showed minimum values at 5-8 degrees C in winter. It was presumed that the formation of NOs by the biotransformation of nonylphenol polyethoxylates decreased much in the sludge treatment of nonionic surfactants at the low temperature (5-8 degrees C) in winter. Average BCF values of NOs and OC in the six kinds of fish were calculated from the field data. The field BCF values of NOs 15-31 in the six kinds of fish were lower than the laboratory BCF values of 167 in Killifish and 282 in Salmon. For OC, the field BCF values 129-297 for the three kinds of fish were nearly equal to the laboratory BCF value, 261, in Killifish. PMID- 10834380 TI - Modelling and prediction of soil sorption coefficients of non-ionic organic pesticides by molecular descriptors. AB - Soil sorption coefficients (K(OC)) of 185 non-ionic organic heterogeneous pesticides have been studied searching for quantitative structure-property relationships (QSPRs). The chemical description of pesticide structure has been made in terms of some molecular descriptors: count descriptors, topological indices, information indices, fragment-based descriptors and weighted holistic invariant molecular (WHIM) descriptors; these last are statistical indices describing size, shape, symmetry and atom distribution of molecules in the three dimensional space. Three new topological indices derived from the electrotopological state indices of Kier and Hall were proposed. Multiple linear regression analysis was performed after previous selection of the descriptors mostly correlated to the response by Genetic Algorithms. The obtained results confirm the capability of the proposed approach to give predictive models for one of the most important partition properties, such as soil sorption coefficient (K(OC)). PMID- 10834381 TI - Chloroperoxidase-mediated chlorination of aromatic groups in fulvic acid. AB - The aim of the present study is to investigate whether exo-enzymatically mediated chlorination of fulvic acid (FA) results in the formation of chlorinated groups within the macromolecules which correspond to those which were previously detected in soil and surface water samples. The chlorination was carried out by exposing FA to a commercial chloroperoxidase (CPO) in the presence of chloride and hydrogen peroxide. The exposed FA was then chemically degraded using an oxidative technique and finally analysed for four different aromatic groups and their chlorinated analogues. The particular aromatic groups included were the methyl esters of 4-ethoxybenzoic acid, 3-methoxy-4-ethoxybenzoic acid, 3,4 diethoxybenzoic acid, and 3,5-dimethoxy-4-ethoxybenzoic acid, along with their mono- and dichlorinated analogues. Prior to the chemical degradation procedure, the FA was analysed for AOX (adsorbable organic halogens) and chlorinated acetic acids. The original FA contained 1.4 mg Cl(org) g(-1) and detectable amounts of two chlorinated aromatic groups. After exposure to the enzyme, the concentration of AOX increased to 44.3 mg Cl(org) g(-1) and detectable amounts of four chlorinated aromatic groups as well as di- and trichloroacetic acid were found. PMID- 10834382 TI - Partitioning of naphthalene to gemini surfactant-treated alumina. AB - Partitioning of naphthalene to anionic surfactants adsorbed on alumina in the aqueous phase was studied for immobilization of the contaminant in the subsurface. Three anionic surfactants with different molecular structures were used: a conventional (sodium dodecylbenzene sulfonate, SDDBS), a gemini (dialkylated disulfonated diphenyl oxide with alkyl chain length of 12, DADS C12), and a dianionic (monoalkylated disulfonated diphenyl oxide with alkyl chain length of 12, MADS-C12). Sorption of the surfactants onto alumina was studied in a series of batch experiments and the effectiveness of the adsorbed surfactants onto aluminum oxide as a sorptive phase for naphthalene was compared. PMID- 10834383 TI - Correlation between neovascularisation and neuroendocrine differentiation in prostatic carcinoma. AB - Neuroendocrine (NE) differentiated tumor cells are found in almost all prostatic carcinomas. Prostatic carcinomas with a high NE differentiation have a poor prognosis and increased metastatic potential. A relationship between the neovascularisation density in the tumor and the metastatic potential in prostatic carcinoma is well known. NE cells and microvessels were demonstrated immunohistochemically on 102 radical prostatectomy specimens using antibodies against Chromogranin A and CD34. Standard areas (7.9 mm2) of maximal Chromogranin A expression and highest vascularisation were determined and topographically related by light microscopy. Area density of microvessels was evaluated morphometrically. NE tumor cells were present in all prostatic carcinomas studied. High grade prostatic carcinomas expressed significantly more NE tumor cells and exhibited a higher neovascularisation than low grade carcinomas. There was significantly higher neovascularisation in high grade tumors with many, as compared to high grade tumors with few, NE tumor cells. Poorer pathological staging correlated with increased neovascularisation and stronger NE differentiation. A topographical relationship between the area of maximal NE tumor cells and the area of highest neovascularisation was found in 80.4% of all cases. An analysis of variance revealed a large number of NE tumor cells as the only predictor of an increased neovascularisation (p = 0.0006). These observations support the concept that increased neovascularisation is influenced not only by poor pathological grading but also by a high NE differentiation. PMID- 10834384 TI - Competence on demand in DNA image cytometry. AB - Quantitation methods in clinical pathology have to be normalized and standardized both from the instrumental and the methodological point of view to guarantee a defined level of precision and accuracy independent of the site where they are applied. The comparability of results obtained in different laboratories is the basis for the application of standardized diagnostic classification systems and therapeutic schemes. Remote quantitation based on standardized evaluation tools could be a way to reach the goals mentioned above. Diagnostic DNA image cytometry, increasingly used as a routine method in clinical pathology, will serve as an example for demonstrating the feasibility and usefulness of a concept of remote quantitation. We report a system for a remote DNA ploidy analysis, based on client server technology, and accessible via Internet or ISDN connections (Quantitation Server EUROQUANT). This system (i) allows the cytometric measurement of the DNA content of cells for diagnostic purposes, (ii) provides the user with comprehensive quality control of such measurements, (iii) helps in trouble-shooting, and (iv) gives assistance in diagnostic interpretation. The system uses the principles of telepathology and Internet technology. To date, more than 40 laboratories from Europe, USA, and Asia have successfully performed analyses on about 3,000 ploidy data sets. PMID- 10834385 TI - Multinucleated giant cells in submucosal layer of human urinary bladder: an immunohistochemical and electron microscopic study. AB - Multinucleated giant cells (MGC) detected in the submucosal layer of human urinary bladder mainly associated with transitional cell carcinoma were examined immunohistochemically and ultrastructurally. The cases examined totaled 29, namely 14 cases with transitional cell carcinoma and another 15 cases mostly with malignancy in other organs. Histologically, MGC were smooth, irregular or dendritic in shape, and tended to increase in number in the vicinity of cancer or marked inflammation. They were consistently positive for not only vimentin, but also MB-2, and CD34, and were mostly positive for proliferating cell nuclear antigen (PCNA), but not MIB-1 (Ki-67) and HLA-DRalpha antigens. On occasion, antibodies to alpha-smooth muscle actin (alpha-SMA), muscle actin (M-actin), CD68 (KP-1) and alpha subunit of S-100 protein also yielded positive reactions. Interestingly, aggregated short bulbous processes were ultrastructurally observed on their surface in parts. These findings suggested that MGC in the submucosal layer of human urinary bladder were MB-2 and CD34-positive multipotential mesenchymal cells with no mitotic activity expressing fibroblastic (vimentin), myofibroblastic (alpha-SMA), or histiocytic (CD68) markers mostly in the vicinity of malignancy, and that these MGC were formed by fusion of mononuclear cells expressing identical markers with those of MGC. Further investigations are needed to clarify the exact function of MGC in human urinary bladder. PMID- 10834386 TI - APC and beta-catenin in alveolar soft part sarcoma (ASPS)--immunohistochemical and molecular genetic analysis. AB - Apart from its role in cell-adhesion, beta-catenin is regarded as an oncoprotein, the cytoplasmic level of which is regulated by APC as a tumor suppressor protein. Changes of chromosome 5q, the region that includes the APC-gene, are known to be important in the pathogenesis of fibromatosis; however, little is known about the significance of APC and beta-catenin in other mesenchymal tumors. Therefore, we used immunohistochemistry and DNA-analysis to investigate four cases of alveolar soft-part sarcoma (ASPS) as a mesenchymal tumor with a distinct histologic appearance. In three cases of ASPS the APC-gene product was found to have strong nuclear expression and only faint cytoplasmic staining. Beta-catenin showed a partly membranous, partly strong intracytoplasmic expression. No gene mutations for APC and beta-catenin were detected in any of the four cases. These investigations suggest that, apart from their function in carcinogenesis and fibromatoses, APC and beta-catenin play a role in the pathogenesis of soft tissue tumors such as ASPS. The significance of a striking nuclear accumulation of non mutated, virtually functionally active APC-tumor suppressor protein has not yet been investigated. A nuclear function of APC in ASPS in down-regulating nuclear transcription processes linked to overexpression of beta-catenin, as is known in colorectal carcinogenesis, may be hypothesized. PMID- 10834387 TI - Effect of antihypertensive drugs on the myocardial microvessels in rats with nitric oxide blockade. AB - In the present study, myocardial microvessels were investigated by stereology in rats with nitric oxide blockade and concomitant antihypertensive treatment for 40 days. The following five groups (10 rats each) were studied: control; L-NAME; L NAME + spironolactone; L-NAME + enalapril; L-NAME + verapamil. The blood pressure (BP) increased every week in the L-NAME group; after an initial increase BP decreased in the treated groups and was not different from the control group. Compared to control animals, the myocardium had hypertrophied myocytes and capillary rarefaction; the tunica media and the tunica intima of small arteries were thickened, and an increase in collagen fibrils in L-NAME treated animals was noted. The enalapril, verapamil and spironolactone groups showed uniform myocardium, quite similar to the control group. The volume density of vessels, in comparison with the L-NAME group, was greater in the spironolactone group (57%), in the enalapril group (76%) and in the verapamil group (81%). The length density of vessels was, respectively, 56%, 50%, and 76% greater in the spironolactone, enalapril and verapamil groups than in the L-NAME group. The surface density of the vessels of the L-NAME group was, respectively, 88%, 96%, and 113% lower than in the spironolactone, enalapril and verapamil groups. These results are compatible with the occurrence of angiogenesis in the verapamil rats. PMID- 10834388 TI - Liver metallothionein expression in thioacetamide-intoxicated rats. AB - Metallothioneins (MT), a group of ubiquitous low molecular weight proteins, implicated primarily in metal ion detoxification, are known to be expressed during hepatocellular proliferation after partial hepatectomy in rats. In the present study, we investigated the expression of MT in a rat model of liver injury and regeneration, induced by intraperitoneal administration of thioacetamide (TAA). The animals were killed at 0, 12, 24, 36, 48, 60, 72, 84, 96, 108 and 120 hours after TAA administration. The rate of tritiated thymidine incorporation into hepatic DNA, the enzymatic activity of thymidine kinase, and the assessment of the mitotic index in hepatocytes were used as indices of liver regeneration. Liver MTs were detected immunohistochemically. TAA administration caused severe hepatic injury, followed by regeneration. MT expression became prominent in hepatocytes as early as 12 hours post-TAA administration. At 24 and 36 hours post-TAA administration intense nuclear and cytoplasmic staining of hepatocytes was found in the vicinity of necrotic areas. The maximal nuclear and cytoplasmic MT expression coincides with the peak of hepatocyte proliferative capacity, occurring at 48 and 60 hours post-TAA administration. MT expression correlated positively with the Zn content of liver tissue, but negatively with serum one, at the time of maximum hepatocyte proliferative capacity. This study suggests that MT participates in hepatocyte replication after toxin-induced liver injury. PMID- 10834389 TI - Detection of the 20-kDa virulence-associated antigen of Rhodococcus equi in malakoplakia-like lesion in pleural tissue obtained from an AIDS patient. AB - A malakoplakia-like lesion was detected in a pleural biopsy from an AIDS patient presenting clinical and radiologic features of pneumonia. Cultures of bronchoalveolar lavage and pleural fluid evidenced Rhodococcus equi as the causative agent of pleuro-pulmonary infection. Immunochemical characterization of the R. equi isolate showed the presence of a strain similar to the ATCC 33704 reference strain presenting the capsular antigen of serotype 4, and the intermediate virulence-associated antigen of 20-kDa. Histopathology of the patient's pleural biopsy showed plaques of macrophages interspersed with lymphocytes, and intracytoplasmic cocci and bacilli in macrophages, which were variably acid-fast positive. Immunohistochemistry of cocci, bacilli and their degradation products resulted strongly positive when stained with a mouse monoclonal antibody (MAb) produced against the 20-kDa antigen. This finding could have important implications for the pathogenicity of R. equi for human beings, since we do not know yet all the factors involved in the formation of malakoplakia. Indeed, the results obtained in the present study, taken together with the results obtained for pigs inoculated with R. equi strains of intermediate virulence (Madarame et al. 1998), raise the possibility that most strains presenting the 20-kDa antigen may be capable of inducing malakoplakia. If this hypothesis is confirmed by immunohistochemical analysis of human pulmonary malakoplakia cases due to R. equi, the detection of this antigen may be extremely helpful in the diagnosis and treatment of such patients. This is the first report of R. equi infection in human beings that suggests a relationship between pleural malakoplakia and the virulence-associated antigen of 20-kDa. PMID- 10834390 TI - Old ectopic pregnancy remnants with morphological features of placental site nodule occurring in fallopian tube and broad ligament. AB - Placental site nodule (PSN) is an asymptomatic benign proliferation of intermediate trophoblast from a previous gestation that failed to involute. It is most commonly found in the endometrium or endocervix; however, placental site nodule has recently been reported to occur at sites of ectopic gestation. This is the first case of PSN in the broad ligament in direct contact with the fallopian tube. The patient underwent surgery for an adenocarcinoma of the opposite tube. Microscopically and immunohistochemically, the lesion showed the characteristics of a proliferation of intermediate trophoblast. PMID- 10834391 TI - Metastatic carcinoma in follicular adenoma of the thyroid gland. AB - We report here two autopsy cases of carcinoma that metastasized to a thyroid follicular adenoma. The first was an 82-year-old man with sigmoid colon adenocarcinoma. Three years after a colectomy he died of multiple metastases. Autopsy revealed that the metastatic colonic carcinoma was located in a thyroid follicular adenoma. The second case was a 51-year-old man with primary lung adenocarcinoma. At the time of admission, he was inoperable because of multiple organ metastasis. He died 14 months after admission due to the carcinoma. Autopsy revealed a metastatic lung adenocarcinoma also located in a follicular adenoma, as in the first case. The phenomenon of tumor-to-tumor metastasis is rare. The mechanism is still unknown, and to our knowledge, only five cases of carcinoma metastatic to thyroid follicular adenoma have been documented in the literature. PMID- 10834392 TI - Sclerosing papillary carcinoma of the thyroid with tubular features. AB - A case of papillary thyroid carcinoma (PTC) in a twenty-seven year old female is reported. The patient presented with complaints of a goiter and associated dysphagia. Histologically, the tumor was characterized by diffuse growth of branching tubules lined by a single layer of the tumor cells and surrounded by circularly arranged fibrous tissue of variable thickness. Focally, micropapillary projections of the epithelium budding into the lumen of tubules were visible. The cells showed features of PTC (overlapping ground glass nuclei, multiple grooves and pseudoinclusions). Nevertheless, no areas of "classical" PTC were identified. A striking feature was the presence of a severe lymphoplasmocytic infiltrate and the presence of plentiful S-100 protein positive dendritic/Langerhans cells. The tumor shares several features with diffuse sclerosing PTC, namely the age and sex of the patient, the infiltrative growth pattern and the presence of severe chronic thyroiditis that is associated with a large number of dendritic/Langerhans cells in the background. There are, however, several distinct differences - particularly the absence of massive squamous metaplasia, psammoma bodies, areas of classical PTC, and also lack of invasion of lymphatic vessels. We believe that this tumor represents an unusual example of diffuse sclerosing PTC with special tubular features. PMID- 10834393 TI - Markers of endothelial dysfunction. AB - The endothelium is a functional barrier between vessel wall and blood stream. Assuming the total human vascular and capillary system occupies a surface area of more than 1,000 m2 which is covered by 1,013 endothelial cells, the complex role of the endothelium for hemostasis and immunological and metabolic processes becomes obvious. Dysfunction of the endothelium is a critical factor in the pathogenesis of vascular diseases and thrombus formation. This paper provides a brief review of physiological endothelial functions and summarizes measurable changes in products released from endothelial cells under pathological conditions which were associated with endothelial dysfunction. PMID- 10834394 TI - Standardization and quality control of PCR analyses. AB - In the very beginning of polymerase chain reaction (PCR) tests entering the field of diagnosis of infectious agents, the introduction of this technology into routine diagnosis was hampered by its frequent tendency to create false-positive results because of contamination. This problem is now widely solved by the introduction of the uracil-N-glycosylase (UNG) anticontamination technology. However, care must still be taken to avoid other sources of producing false positive results. They might additionally derive from human error and/or insufficient PCR amplification and detection protocols. A special case lies in the fact that PCR also amplifies DNA from dead organisms rendering a result diagnostically correct as positive, but clinically as false-positive. In PCR, as in any other diagnostic test, the risk of creating a false-negative result also exists. In such a case, the most probable source besides human error, low target or poor amplification and detection protocols is an inhibition caused by interfering substances in a patient's sample. Strategies to recognize and overcome this issue are discussed in this article. Finally a few results from quality control studies on amplification technologies in the diagnosis of infectious agents are reviewed. PMID- 10834395 TI - Histochemical and immunocytochemical approaches to the study of oxidative stress. AB - We review an array of newly developed in situ detection methods that can be used for the qualitative and semi-quantitative measurement of various indices related to oxidative stress. The importance of in situ methods over bulk analysis cannot be overstated when considering the structural and cellular complexity of tissue and the effects of diseases thereof. Indeed, in situ detection allows detection of specific cell types affected or specific localization such that a process affecting only a small fraction of the tissue or cells can be readily visualized. Consequently, a positive signal in situ indicates real levels that cannot be masked by unrelated or compensatory responses in adjacent cells, and corrections can be easily made for the modifications to long-lived proteins during physiological aging. In fact, the damage to extracellular matrix proteins of major vessels, provides a cumulative record of long-term oxidative insult. Yet the same properties that make vessels ideal markers for aging limits their sensitivity to detect disease-specific changes unless in situ techniques are used. PMID- 10834396 TI - Sniffing out the truth: clinical diagnosis using the electronic nose. AB - Recently the use of smell in clinical diagnosis has been rediscovered due to major advances in odour sensing technology and artificial intelligence (AI). It was well known in the past that a number of infectious or metabolic diseases could liberate specific odours characteristic of the disease stage. Later chromatographic techniques identified an enormous number of volatiles in human clinical specimens that might serve as potential disease markers. "Artificial nose" technology has been employed in several areas of medical diagnosis, including rapid detection of tuberculosis (TB), Helicobacter pylori (HP) and urinary tract infections (UTI). Preliminary results have demonstrated the possibility of identifying and characterising microbial pathogens in clinical specimens. A hybrid intelligent model of four interdependent "tools", odour generation "kits", rapid volatile delivery and recovery systems, consistent low drift sensor performance and a hybrid intelligent system of parallel neural networks (NN) and expert systems, have been applied in gastric, pulmonary and urine diagnosis. Initial clinical tests have shown that it may be possible in the near future to use electronic nose technology not only for the rapid detection of diseases such as peptic ulceration, UTI, and TB but also for the continuous dynamic monitoring of disease stages. Major advances in information and gas sensor technology could enhance the diagnostic power of future bio-electronic noses and facilitate global surveillance models of disease control and management. PMID- 10834398 TI - Paraneoplastic syndromes of the nervous system. AB - Paraneoplastic syndromes affecting the nervous system are unique among immune mediated disorders in that the trigger of the immune response is known: tumor expression of proteins normally restricted to neurons (or other immunoprivileged sites, such as testis) but ectopically expressed in some cancers results in an immunological response characterized by high titers of antibodies targeting the "onconeuronal" antigen. A T-cell response is also elicited in some paraneoplastic syndromes and may be the cause of neuronal destruction. In some instances genes that code for the antigens recognized by the autoantibodies have been identified, cloned and sequenced. Some of the proteins so identified are RNA binding proteins but their specific function has not been identified. In some individuals with cancer but no paraneoplastic syndrome, low titers of antibody can be identified in the serum. Low titers of antibody are associated with a better prognosis of the cancer. Experimental animals immunized against a paraneoplastic antigen are partially protected against tumors that express that antigen. PMID- 10834399 TI - Genetic modifying factors in beta-thalassemia. AB - The beta-thalassemia is probably the most extensively studied genetic disease. Essentially any molecular defect that has been first described in association with the globin genes has been later implicated as a molecular determinant of newly discovered genes. Accordingly, the thalassemias have always represented a model genetic disease, especially in relation to the development of programs for population screening, genetic counseling and prenatal diagnosis. Here we will review the present knowledge on the genetics of thalassemia and of the relevant modifying factors. Major categories of the carrier state, the genotypes, the clinical phenotypes and the correlation between genotype and phenotype will be discussed. PMID- 10834397 TI - Oncogenes and thyroid cancer. AB - Human thyroid tumors can be derived either from epithelial follicular cells or from parafollicular C-cells. Follicular cell-derived tumors represent a wide spectrum of lesions, ranging from benign adenomas through differentiated (follicular and papillary) and undifferentiated (anaplastic) carcinomas, thus providing a good model for finding a correlation between specific genetic lesions and histologic phenotype. Follicular adenomas and carcinomas show frequently the presence of mutations in one of the three ras genes. Papillary carcinomas show frequently a specific gene rearrangement which gives rise to the formation of several types of so-called RET/PTC chimeric genes. This lesions occur in almost 50% of papillary cancers and consist in the juxtaposition of the 3' or tyrosine kinase domain of the RET gene (which codes for a receptor protein not normally expressed in follicular thyroid cells) with the 5' domain of ubiquitously expressed genes, which provide the promoter and dimerization functions, necessary for the constitutive activation of RET/PTC proteins. Anaplastic carcinomas are frequently associated with mutations of the p53 tumor suppressor. Finally, point mutations of the RET gene are found in familial endocrine syndromes (FMTC; MEN2A and MEN2B), a common feature of which is the medullary thyroid carcinoma, a malignant tumor derived from parafollicular C-cells. PMID- 10834400 TI - The genetics of multiple sclerosis. AB - Classical genetic studies involving the analysis of pedigrees and recurrence risk within families have defined the extent to which genetic factors contribute to the aetiology of multiple sclerosis. Limited progress has been made in identifying the number and topography of genetic loci contributing to susceptibility through molecular investigation either of candidate genes or the whole genome using microsatellite and single nucleotide polymorphisms. PMID- 10834401 TI - Near-infrared reflectance spectroscopy for noninvasive monitoring of metabolites. AB - An important class of substances in clinical chemistry are metabolites in body fluids, which are accessible by near-infrared spectroscopy without sample treatment using reagentless, fast and readily automated in vitro assays. Furthermore, noninvasive sensing systems are under development for the determination of blood glucose, especially for diabetic patients or for monitoring in intensive care and surgery. Near-infrared diffuse reflectance spectrometry of skin was employed allowing a certain tissue volume to be integrally probed. For calibration, the partial least-squares (PLS) algorithm was used either based on wide spectral intervals or using special spectral variable selection. Capillary blood glucose reference concentrations were obtained by finger pricking and an automated laboratory method (hexokinase/G6P-DH). Clear evidence is provided for the physical effect, as manifested by the spectral glucose absorptivities, underlying the individual single-person calibration models, which still require improvements in the methodology in the normo- and hypoglycemic concentration range. In extending the potential of noninvasive blood assays by infrared spectroscopy, a novel technique is presented for probing the intravascular fluid space by using fast spectral near-infrared measurements of skin tissue. The pulsatile blood spectrum can be derived from reflectance spectra of oral mucosa by Fourier analysis (near-infrared plethysmography). Future applications and prospects for noninvasive blood assays are discussed. PMID- 10834402 TI - Ribozyme gene therapy for autosomal dominant retinal disease. AB - Gene delivery to cells of the retina, particularly to photoreceptor cells, has broad potential both for answering basic questions of retinal biology and for more applied therapeutic purposes. The use of ribozymes as therapy for autosomal dominant retinal diseases is a promising technique, and the theoretical and practical basis for their use is discussed. The process involves designing and testing ribozymes first in vitro and then in animal models of retinal disease. Viral vectors based on the nonpathogenic human adeno-associated virus, when coupled with the strong, rod photoreceptor specific opsin promoter, offer an efficient and nontoxic way to deliver and express ribozymes in photoreceptor cells for long time periods of time. Effective ribozyme-mediated therapy also demands careful in vitro analysis of a ribozyme's ability to efficiently and specifically distinguish between mutant and wild type RNAs. Finally, effective demonstration of therapy in an animal model requires careful analysis of any rescue effect in the retina using multiple criteria, including biochemical, structural and physiological assays. For this purpose, ribozyme therapy in a transgenic rat model of retinitis pigmentosa containing a dominant rod opsin mutation (proline-to-histidine change at position 23) is discussed in detail. PMID- 10834404 TI - Experiences with external quality assessment (EQA) in molecular diagnostics in clinical laboratories in Germany. Working Group of the German Societies for Clinical Chemistry (DGKC) and Laboratory Medicine (DGLM). AB - The German Societies for Clinical Chemistry (DGKC) and Laboratory Medicine (DGLM) have established an official working group on molecular diagnostics in the field of laboratory medicine. The group's objectives are to support the establishment of molecular biology methods for the use in diagnostics in German clinical laboratories. Towards this end, we have defined specific aims, which are 1) the implementation and extension of external quality assessment (EQA) schemes and methodological exercises offered to clinical diagnostic laboratories, 2) the establishment of a proficiency network and data base within the societies, 3) the implementation of an educational program in molecular diagnostic procedures for clinical chemists and laboratory physicians through the organisation of symposia and workshops and 4) the cooperation with other DGKC/DGLM working groups on shared aspects of laboratory analysis, e.g. preanalytics. The focus of this presentation is to introduce some of these goals in more detail with particular emphasis on the first two program aspects and to discuss experiences with these activities. PMID- 10834403 TI - Oxidized lipoproteins and endothelium. AB - Endothelium is an early target of pro-atherosclerotic events, which may result in functional and morphological perturbations. Oxidized low density lipoproteins, an atherogenic factor with strong cytotoxicity, may potentially contribute to altered endothelial function through the activation of a stress response, which would rescue cells to full vitality, or, conversely, by leading to cell death. Evidence is presented here for the ability of chemically oxidized low density lipoproteins to induce the synthesis of the inducible form of heat shock protein 70 in cultured human endothelial cells, and for the association of epitopes of these modified lipoproteins with apoptotic endothelial cells in aortic sections from hypercholesterolemic rabbits. PMID- 10834405 TI - Unravelling genetic data by arrayed primer extension. AB - We have developed a method for arrayed primer extension (APEX) on an oligonucleotide microchip together with the 4-color fluoresence imaging equipment and supporting software, that allows analysis of the DNA sequence and changes in it. Mutation analysis of BRCA1 gene and single nucleotide polymorphism (SNP) chip for genotyping were used as a model system. Chip surface chemistry, template preparation and APEX reaction conditions were optimised and the assay is ready to be implemented in variety of DNA analysis from SNP testing to DNA resequencing. PMID- 10834406 TI - Evaluation of RNA isolation methods and reference genes for RT-PCR analyses of rare target RNA. AB - Reverse transcription polymerase chain reaction (RT-PCR) analysis is increasingly becoming part of the diagnostic and prognostic evaluation for hematologic and oncologic disorders. Currently, different RNA isolation methods are used in the diagnostic laboratories. No data are available on their suitability for sensitive detection of breakpoint cluster region-abelson (BCR-ABL) gene transcripts. We have extracted RNA from mononuclear cell (MNC) fractions and from lysed blood samples of 4 patients (1 with leukocytosis, 1 with chronic myelogeneous leukemia (CML) under interferon treatment, and 2 CML patients after bone marrow transplantation) with 3 RNA isolation reagents (TRIzol, RNAzol, FastTube reagent). RNA yield was slightly higher with RNAzol than with TRIzol as indicated by agarose gel electrophoresis and spectrophotometric measurement at 260 nm. The FastTube reagent was unsuitable for RNA isolation from MNC, and was not evaluated for lysed blood. Quantitative competitive RT-PCR amplification of the ABL gene showed comparable results for RNA isolated with RNAzol and TRIzol. In RNA samples extracted from lysed whole blood, the presence of amplifiable RNA/cDNA was confirmed by amplification of 4 selected reference genes (porphobilinogen deaminase (PBGD), ABL, the gene spanning the BCR on chromosome 22 and retinoic acid receptor alpha (RARA)) in a multiplex PCR. High quality, DNA-free RNA was obtained with RNAzol, and 1 BCR-ABL-positive (specific for translocation t [9; 221) cell among 2x10(4) normal cells was successfully detectable by single step RT-PCR. In RNA isolated with TRIzol, major contaminations with genomic DNA were observed which significantly impaired the interpretation of the results of RT-PCR analysis. PMID- 10834407 TI - IFCC WorldLab daily news. International Federation of Clinical Chemistry and Laboratory Medicine. PMID- 10834408 TI - Impaired glucose tolerance: the irrepressible alpha-cell? PMID- 10834409 TI - Nighttime continuous subcutaneous insulin infusion revisited: a strategy for improving insulin delivery. PMID- 10834410 TI - Therapeutic importance of subsets of type 2 diabetes? PMID- 10834411 TI - Insulin glargine (HOE901): first responsibilities: understanding the data and ensuring safety. PMID- 10834412 TI - Use of insulin pump therapy at nighttime only for children 7-10 years of age with type 1 diabetes. AB - OBJECTIVE: Because of age-related developmental and cognitive issues, children <10 years of age may not be able to wear an insulin pump safely when they are not under direct parental supervision. The purpose of this study was to determine if insulin pump therapy at nighttime only, when children are at home, could improve fasting and nighttime blood glucose levels without adverse effects. RESEARCH DESIGN AND METHODS: The study cohort consisted of 10 children aged 7-10 years. A randomized crossover design was used to compare nighttime-only pump usage from dinner and throughout the night, combined with a prebreakfast injection of intermediate-acting NPH and rapid-acting lispro insulin, with 3 insulin injections per day. Comparisons were made among mean blood glucose values and percentage of blood glucose levels within the target range (70-150 mg/dl) before meals, at bedtime, and at 3:00 A.M.; serum fructosamine levels; and scores on measures of adherence and fear of hypoglycemia. RESULTS: Compared with baseline levels, the use of the pump resulted in a significant decrease in the mean average (P < 0.001), breakfast (P < 0.0001), and 3:00 A.M. (P < 0.003) blood glucose levels. There was a decrease in the percentage of blood glucose values less than the target range (P < 0.01) and in fructosamine (P < 0.01) values and an increase in the percentage of blood glucose levels within the target range (P < 0.03). CONCLUSIONS: Nighttime-only insulin pump therapy may be a viable alternative that young children can use to improve glycemia when they are not capable of independently managing an insulin pump. PMID- 10834413 TI - Use of insulin aspart, a fast-acting insulin analog, as the mealtime insulin in the management of patients with type 1 diabetes. AB - OBJECTIVE: To compare long-term glycemic control and safety of using insulin aspart (IAsp) with that of regular human insulin (HI). RESEARCH DESIGN AND METHODS: This was a multicenter randomized open-label 6-month study (882 subjects) with a 6-month extension period (714 subjects) that enrolled subjects with type 1 diabetes. Subjects administered IAsp immediately before meals or regular HI 30 min before meals; basal NPH insulin was taken as a single bedtime dose in the majority of subjects. Glycemic control was assessed with HbA1c values and 8-point blood glucose profiles at 3-month intervals. RESULTS: Mean postprandial blood glucose levels (mg/dl +/- SEM) were significantly lower for subjects in the IAsp group compared with subjects in the HI group after breakfast (156 +/- 3.4 vs. 185 +/- 4.7), lunch (137 +/- 3.1 vs. 162 +/- 4.1), and dinner (153 +/- 3.1 vs. 168 +/- 4.1), when assessed after 6 months of treatment. Mean HbA1c values (% +/- SEM) were slightly, but significantly, lower for the IAsp group (7.78% +/- 0.03) than for the regular HI group (7.93% +/- 0.05, P = 0.005) at 6 months. Similar postprandial blood glucose and HbA1c values were observed at 12 months. Adverse events and overall hypoglycemic episodes were similar for both treatment groups. CONCLUSIONS: Postprandial glycemic control was significantly better with IAsp compared with HI after 6 and 12 months of treatment. The improvement was not obtained at an increased risk of hypoglycemia. HbA1c was slightly, but significantly, lower for IAsp compared with HI at 6 and 12 months. PMID- 10834414 TI - Consequences of a family history of type 1 and type 2 diabetes on the phenotype of patients with type 2 diabetes. AB - OBJECTIVE: To investigate the impact of a family history of type 1 and type 2 diabetes on the phenotype of patients with type 2 diabetes. RESEARCH DESIGN AND METHODS: In a population-based study, we compared the phenotype in 3 groups of patients with type 2 diabetes. The first group had no family history of diabetes (FH-, n = 148); the second group had a family history of type 2 diabetes only (FH+(TYPE2), n = 1,211); and the third group had a family history of both type 1 and type 2 diabetes (FH+(MIXED), n = 240). Furthermore, we studied the frequency of GAD antibodies (GADabs), HLA-DQB1 risk genotypes, and the presence of coronary heart disease (CHD) according to family history in unrelated patients with type 2 diabetes from 787 families (148 FH-, 546 FH+(TYPE2) and 93 FH+(MIXED)). RESULTS: Patients with no family history of diabetes were older at the onset of the disease, had a better beta-cell function (P = 0.004), and had higher HDL cholesterol concentrations (P = 0.006) than patients with a family history of diabetes. Patients with a family history of only type 2 diabetes had higher BMI and fasting C-peptide concentrations (P = 0.031) but lower frequency of GADab (11 vs. 23%, P = 0.001) and DQB1 risk genotypes (37 vs. 54%, P = 0.003) compared with patients with a family history of both type 1 and type 2 diabetes. In addition, hypertension (P = 0.05) and CHD (P = 0.031) were more common in FH+(TYPE2) than in FH+(MIXED) patients. In patients <60 years old, a family history of type 1 diabetes was associated with a reduced risk of CHD independent of age, hypertension, and HDL cholesterol concentrations. The results were similar when the GADab+ patients were excluded from the analysis. CONCLUSIONS: A family history of both type 1 and type 2 diabetes had a profound influence on the phenotype of patients with type 2 diabetes, which suggests a genetic interaction between type 1 and type 2 diabetes. PMID- 10834415 TI - Diabetes in the Old Order Amish: characterization and heritability analysis of the Amish Family Diabetes Study. AB - OBJECTIVE: The Old Order Amish (OOA) are a genetically well-defined closed Caucasian founder population. The Amish Family Diabetes Study was initiated to identify susceptibility genes for type 2 diabetes. This article describes the genetic epidemiology of type 2 diabetes and related traits in this unique population. RESEARCH DESIGN AND METHODS: The study cohort comprised Amish probands with diabetes who were diagnosed between 35 and 65 years of age and their extended adult family members. We recruited 953 adults who represented 45 multigenerational families. Phenotypic characterization included anthropometry, blood pressure, diabetes status, lipid profile, and leptin levels. RESULTS: The mean age of study participants was 46 years, and the mean BMI was 26.9 kg/m2. Subjects with type 2 diabetes were older, more obese, and had higher insulin levels. The prevalence of diabetes in the OOA was approximately half that of the Caucasian individuals who participated in the Third National Health and Nutrition Examination Survey (95% CI 0.23-0.84). The prevalence of diabetes in the siblings of the diabetic probands was 26.5% compared with a prevalence of 7.0% in spouses (lambdaS = 3.28, 95% CI 1.58-6.80). The heritability of diabetes-related quantitative traits was substantial (13-70% for obesity-related traits, 10-42% for glucose levels, and 11-24% for insulin levels during the oral glucose tolerance test; P = 0.01 to <0.0001). CONCLUSIONS: Type 2 diabetes in the Amish has similar phenotypic features to that of the overall Caucasian population, although the prevalence in the Amish community is lower than that of the Caucasian population. There is significant familial clustering of type 2 diabetes and related traits. This unique family collection will be an excellent resource for investigating the genetic underpinnings of type 2 diabetes. PMID- 10834416 TI - Sex differentials in predictors of mortality for patients with adult-onset diabetes: a population-based follow-up study in Beer-Sheva, Israel. AB - OBJECTIVE: To test the hypothesis that factors predicting mortality differ between diabetic men and women. RESEARCH DESIGN AND METHODS: A total of 498 known patients with diabetes residing in a well-defined geographical area and receiving primary health care in 3 primary care community clinics were interviewed and examined between 1988 and 1990. RESULTS: By 31 July 1998, after a mean follow-up period of 7.8 years, 148 patients (68 men and 80 women) had died (29.7%). No statistical differences in survival rate or in the specific causes of death were found between men and women. In the univariate analysis of factors examined at baseline, GHb levels were significantly higher among women who died compared with women who survived, but this was not the case for men. Conversely, a trend of higher triglyceride and uric acid levels was found for men who died compared with men who survived, but this was not the case for women. Multivariate Poisson regression analysis showed significantly higher risk ratios for mortality in men > or = 63 years of age, men with microalbumin excretion > or = 30 mg/l, and men with higher triglyceride levels. In contrast, the analysis in women showed that higher GHb and creatinine levels and a reported history of heart disease were the only factors at the baseline examination significantly and independently associated with an increased risk ratio of mortality. CONCLUSIONS: The results suggest the existence of sex-specific interactions with various metabolic factors associated with diabetes that may have a different effect on mortality for each sex. PMID- 10834417 TI - Screening techniques to identify people at high risk for diabetic foot ulceration: a prospective multicenter trial. AB - OBJECTIVE: Diabetic foot ulceration is a preventable long-term complication of diabetes. A multicenter prospective follow-up study was conducted to determine which risk factors in foot screening have a high association with the development of foot ulceration. RESEARCH DESIGN AND METHODS: A total of 248 patients from 3 large diabetic foot centers were enrolled in a prospective study. Neuropathy symptom score, neuropathy disability score (NDS), vibration perception threshold (VPT), Semmes-Weinstein monofilaments (SWFs), joint mobility, peak plantar foot pressures, and vascular status were evaluated in all patients at the beginning of the study. Patients were followed-up every 6 months for a mean period of 30 months (range 6-40), and all new foot ulcers were recorded. The sensitivity, specificity, and positive predictive value of each risk factor were evaluated. RESULTS: Foot ulcers developed in 95 feet (19%) or 73 patients (29%) during the study. Patients who developed foot ulcers were more frequently men, had diabetes for a longer duration, had nonpalpable pedal pulses, had reduced joint mobility, had a high NDS, had a high VPT, and had an inability to feel a 5.07 SWE NDS alone had the best sensitivity, whereas the combination of the NDS and the inability to feel a 5.07 SWF reached a sensitivity of 99%. On the other hand, the best specificity for a single factor was offered by foot pressures, and the best combination was that of NDS and foot pressures. Univariate logistical regression analysis yielded a statistically significant odds ratio (OR) for sex, race, duration of diabetes, palpable pulses, history of foot ulceration, high NDSs, high VPTs, high SWFs, and high foot pressures. In addition, 94 (99%) of the 95 ulcerated feet had a high NDS and/or SWF which resulted in the highest OR of 26.2 (95% CI 3.6-190). Furthermore, in multivariate logistical regression analysis, the only significant factors were high NDSs, VPTs, SWFs, and foot pressures. CONCLUSIONS: Clinical examination and a 5.07 SWF test are the two most sensitive tests in identifying patients at risk for foot ulceration, especially when the tests are used in conjunction with each other. VPT measurements are also helpful and can be used as an alternative. Finally, foot pressure measurements offer a substantially higher specificity and can be used as a postscreening test in conjunction with providing appropriate footwear. PMID- 10834418 TI - Impact of diabetes on crash risks of truck-permit holders and commercial drivers. AB - OBJECTIVE: The U.S. and some Canadian government agencies have waived commercial license restrictions for some insulin-using diabetic drivers. However, the U.S. Federal Highway Administration is no longer giving waivers. Scientific evidence to support such regulations has been sparse. This article presents detailed analyses of crash risks for users and nonusers of insulin among diabetic truck permit holders in Quebec, Canada. RESEARCH DESIGN AND METHODS: Diabetic truck permit holders were group-matched by age to a random sample of healthy permit holders. Data on permits, medical conditions, and crashes involving 13,453 permit holder-years in 1987-1990 were extracted from the files of the public insurer for automobile injuries in Quebec. Additional health status data were obtained from the provincial public health insurer. A telephone survey was conducted to collect data on driving patterns and exposure. Risk ratios were estimated using negative binomial regression models. RESULTS: Risk ratios for crashes vary by category of diabetes. Permit holders for single-unit trucks (STs) who are diabetic without complications and not using insulin have an increased crash risk of 1.68 when compared with healthy permit holders of the same permit class. When controlling for risk exposure, commercial drivers with an ST permit and the same diabetic condition have an increased risk of 1.76. Insulin use is not associated with higher crash risk. CONCLUSIONS: The increased crash risk for the group with uncomplicated diabetes not using insulin is a new finding. The lack of consistent increases in crash risks among diabetic commercial drivers with complications or who use insulin may be a "healthy worker effect" masking the real risk, because these licensees have a lower participation rate as professional drivers. PMID- 10834419 TI - Fluoxetine for depression in diabetes: a randomized double-blind placebo controlled trial. AB - OBJECTIVE: Depression is prevalent in patients with diabetes. It is associated with poor glycemic control and is linked to an increased risk for diabetic complications. In this study, we assessed the efficacy of fluoxetine for depression in patients with diabetes. RESEARCH DESIGN AND METHODS: Sixty patients with diabetes (type 1, n = 26; type 2, n = 34) and major depressive disorder entered an 8-week randomized placebo-controlled double-blind trial. Patients were given daily doses of fluoxetine (up to 40 mg/day). The Beck Depression Inventory (BDI) and Hamilton Rating Scale for Depression (HAMD) were used to measure the severity of depression and to determine the percentage of patients who achieved substantial improvement or complete remission. GHb levels were obtained to monitor glycemic control. RESULTS: Reduction in depression symptoms was significantly greater in patients treated with fluoxetine compared with those receiving placebo (BDI, -14.0 vs. -8.8, P = 0.03; HAMD, -10.7 vs. -5.2, P = 0.01). The percentage of patients achieving a significant improvement in depression per the BDI was also higher in the fluoxetine group (66.7 vs. 37.0%, P = 0.03). Additionally, trends toward a greater rate of depression remission (48.1 vs. 25.9%, P = 0.09 per the HAMD) and greater reduction in GHb (-0.40 vs. -0.07%, P = 0.13) were observed in the fluoxetine group. CONCLUSIONS: Fluoxetine effectively reduces the severity of depression in diabetic patients. Our study demonstrated that after only 8 weeks, this treatment also produced a trend toward better glycemic control. PMID- 10834420 TI - Heart rate in relation to insulin sensitivity and insulin secretion in nondiabetic subjects. AB - OBJECTIVE: Elevated heart rate has been predictive of cardiovascular disease and has been proposed as a global index of the autonomic nervous system influence on the heart. Hyperinsulinism has been shown to trigger sympathetic activity experimentally; however, the clinical and epidemiological data on the association of heart rate with hyperinsulinism and insulin resistance are conflicting. RESEARCH DESIGN AND METHODS: Insulin sensitivity (S(I)) and the acute insulin response (AIR) to glucose were assessed by a frequently sampled intravenous glucose tolerance test and related to resting heart rate in the tri-ethnic nondiabetic population (n = 1,000) of the Insulin Resistance Atherosclerosis Study. RESULTS: Heart rate was related to fasting insulin (r = 0.20), intact proinsulin (r = 0.15), split proinsulin (r = 0.17), and AIR (r = 0.18), and an inverse relation was found between heart rate and S(I) (r = -0.19) (all P values <0.0001, adjusted for age, sex, ethnicity, glucose tolerance status, and smoking). In a multiple linear regression analysis (adjusting for age, sex, ethnicity, clinical center, glucose tolerance status, and smoking), heart rate was significantly and independently associated with AIR, proinsulin, and S(I). CONCLUSIONS: Proinsulin, acute insulin secretion, and S(I) are associated with heart rate in nondiabetic subjects. PMID- 10834421 TI - Optimized basal-bolus therapy using a fixed mixture of 75% lispro and 25% NPL insulin in type 1 diabetes patients: no favorable effects on glycemic control, physiological responses to hypoglycemia, well-being, or treatment satisfaction. AB - OBJECTIVE: To investigate the effects of a multiple injection regimen with a mixture of 75% lispro and 25% intermediate-acting insulin (lispro high mixture [HM]) before meals on glycemic control, physiological responses to hypoglycemia, well-being, and treatment satisfaction. RESEARCH DESIGN AND METHODS: We studied 35 type 1 diabetes patients. After an 8- to 10-week lead-in period, patients were randomized to HM or human regular insulin therapy for 12-14 weeks. During the lead-in and treatment periods, HbA1c levels and hypoglycemic frequencies were measured, and patients completed the Well-Being Questionnaire and the Diabetes Treatment Satisfaction Questionnaire. In 19 patients, responses to hypoglycemia were tested during stepped euglycemic-hypoglycemic clamps. RESULTS: HM treatment improved postprandial glycemia but had no effect on HbA1c, frequency of hypoglycemia, well-being, or treatment satisfaction. During experimental hypoglycemia, HM therapy was associated with a slightly lower total adrenaline response and a higher autonomic symptom threshold (i.e., the autonomic symptom response occurred at a lower blood glucose level) than human regular insulin therapy. We speculate that this effect resulted from an accumulation of insulin during the night. CONCLUSIONS: Multiple injection therapy with HM rather than human regular insulin before meals does not offer advantages regarding glycemic control, frequency of hypoglycemia, well-being, or treatment satisfaction. In addition, this regimen causes an attenuation of the adrenaline and autonomic symptom responses to hypoglycemia. PMID- 10834422 TI - Efficacy of injected liquid silicone in the diabetic foot to reduce risk factors for ulceration: a randomized double-blind placebo-controlled trial. AB - OBJECTIVE: To investigate the effectiveness of injecting liquid silicone in the diabetic foot to reduce risk factors for ulceration in a randomized double-blind placebo-controlled trial. RESEARCH DESIGN AND METHODS: A total of 28 diabetic neuropathic patients without peripheral vascular disease were randomized to active treatment with 6 injections of 0.2 ml liquid silicone in the plantar surface of the foot or to treatment with an equal volume of saline (placebo). No significant differences were evident regarding age or neuropathy status between the 2 groups. All injections were under the metatarsal heads at sites of calluses or high pressures. Barefoot plantar pressures (pedobarography) and plantar tissue thickness under the metatarsal heads (Planscan ultrasound device) were measured at baseline and at 3, 6, and 12 months after the first injection. Injection sites were photographed at all stages, and callus formation was scored as a change from baseline. Throughout the study, patients were treated by the same podiatrist for all podiatry treatment. RESULTS: Patients who received silicone treatment had significantly increased plantar tissue thickness at injection sites compared with the placebo group (1.8 vs. 0.1 mm) (P < 0.0001) and correspondingly significantly decreased plantar pressures (-232 vs. -25 kPa) (P < 0.05) at 3 months, with similar results at 6 and 12 months. A trend was noted toward a reduction of callus formation in the silicone-treated group compared with no change in the placebo group. CONCLUSIONS: The results confirm the efficacy of plantar silicone injections in reducing recognized risk factors associated with diabetic foot ulceration. PMID- 10834423 TI - Less hypoglycemia with insulin glargine in intensive insulin therapy for type 1 diabetes. U.S. Study Group of Insulin Glargine in Type 1 Diabetes. AB - OBJECTIVE: Insulin glargine (21A-Gly-30Ba-L-Arg-30Bb-L-Arg-human insulin) is a biosynthetic insulin analog with a prolonged duration of action compared with NPH human insulin. This study compared insulin glargine with NPH human insulin in subjects with type 1 diabetes who had been previously treated with multiple daily injections of NPH insulin and regular insulin. RESEARCH DESIGN AND METHODS: This study was a multicenter randomized parallel-group study in which subjects were randomized to receive premeal regular insulin and either insulin glargine (at bedtime) or NPH insulin (at bedtime for patients on once-daily therapy and at bedtime and in the morning for patients on twice-daily therapy) for up to 28 weeks. Dose titration of both basal insulins was based on capillary fasting whole blood glucose (FBG) levels; the goal was a premeal blood glucose concentration of 4.4-6.7 mmol/l. RESULTS: A total of 534 well-controlled type 1 diabetic subjects (mean GHb 7.7%, mean fasting plasma glucose [FPG] 11.8 mmo/l) were treated. A small decrease in GHb levels was noted with both insulin glargine (-0.16%) and NPH insulin (-0.21%; P > 0.05). Significant reductions in median FPG levels from baseline (-1.67 vs. -0.33 mmol/l with NPH insulin, P = 0.0145) and a trend for a reduction in capillary FBG levels were achieved with insulin glargine. After the 1-month titration phase, significantly fewer subjects receiving insulin glargine experienced symptomatic hypoglycemia (39.9 vs. 49.2%, P = 0.0219) or nocturnal hypoglycemia (18.2 vs. 27.1%, P = 0.0116) with a blood glucose level <2.0 mmol/l compared with subjects receiving NPH insulin. CONCLUSIONS: Lower FPG levels with fewer episodes of hypoglycemia were achieved with insulin glargine compared with once- or twice-daily NPH insulin as part of a basal-bolus regimen in patients with type 1 diabetes. PMID- 10834424 TI - Time-action profile of the long-acting insulin analog insulin glargine (HOE901) in comparison with those of NPH insulin and placebo. AB - OBJECTIVE: To study the pharmacodynamic properties of the subcutaneously injected long-acting insulin analog HOE901 (30 microg/ml zinc) in comparison with those of NPH insulin and placebo. RESEARCH DESIGN AND METHODS: In this single-center double-blind euglycemic glucose clamp study, 15 healthy male volunteers (aged 27 +/- 4 years, BMI 22.2 +/- 1.8 kg/m2) received single subcutaneous injections of 0.4 U/kg body wt of HOE901, NPH insulin, or placebo on 3 study days in a randomized order. The necessary glucose infusion rates (GIRs) to keep blood glucose concentrations constant at 5.0 mmol/l were determined over a 30-h period after administration. RESULTS: The injection of HOE901 did not induce the pronounced peak in metabolic activity observed with NPH insulin (GIRmax 5.3 +/- 1.1 vs. 7.7 +/- 1.3 mg x kg(-1) x min(-1)) (P < 0.05); after an initial rise, metabolic activity was rather constant over the study period. This lack of peak was confirmed by a lower glucose consumption in the first 4 h after injection (area under the curve from 0 to 4 h [AUC(0-4 h)] 1.02 +/- 0.34 vs. 1.48 +/- 0.34 g/kg) (P < 0.001) with HOE901, as compared with NPH insulin. In this single-dose study, the metabolic effect measured over a period of 30 h was lower with HOE901 than with NPH insulin (AUC(0-30 h) 7.93 +/- 1.82 vs. 9.24 +/- 1.29 g/kg) (P < 0.05). CONCLUSIONS: This study shows that the soluble long-acting insulin analog HOE901 induces a smoother metabolic effect than NPH insulin, from which a better substitution of basal insulin requirements may follow. PMID- 10834425 TI - Islet dysfunction in insulin resistance involves impaired insulin secretion and increased glucagon secretion in postmenopausal women with impaired glucose tolerance. AB - OBJECTIVE: To characterize in detail the association between insulin sensitivity and islet function in relation to glucose tolerance in nondiabetic subjects. RESEARCH DESIGN AND METHODS: The study included 108 postmenopausal women, aged 57 59 years, with normal glucose tolerance (NGT) or impaired glucose tolerance (IGT) and measured glucose tolerance (World Health Organization, 75 g glucose), insulin sensitivity (euglycemic-hyperinsulinemic clamp), and islet function (the 2-5 min insulin responses [AIR] and glucagon [AGR] responses to 5 g intravenous arginine at fasting, 14 and >25 mmol/l glucose levels). The product of insulin sensitivity and secretion was calculated (disposition index [DI]) and used to study the relationship between the two parameters. RESULTS: Insulin sensitivity and insulin secretion were highly inversely correlated in a hyperbolic manner (r > 0.64, P < 0.001) in women with NGT (n = 71). Women with IGT (n = 37) had reduced insulin sensitivity compared with women with NGT (P = 0.011). The AIRs were not appropriately increased in relation to the reduced insulin sensitivity in the IGT women, demonstrated as reduced DI in IGT compared with NGT (P < 0.001). Further, women with IGT had an increased AGR (P < 0.001) and a reduced glucose inhibition of glucagon secretion (slopeAGR, P = 0.014) compared with women with NGT. In a multivariate regression model including all of the 108 women, 2-h glucose was independently determined by the DI, the AGR, and the slopeAGR (r = 0.63, P < 0.001). CONCLUSIONS: We have shown that both the individual ability to adapt insulin secretion to the ambient insulin sensitivity and the level of glucagon secretion are important parameters for maintenance of NGT. Therefore, islet dysfunction in IGT involves low insulin and high glucagon secretion, which present potential targets for correcting impaired glycemia. PMID- 10834426 TI - Hemodynamic effects of acute hyperglycemia in type 2 diabetic patients. AB - OBJECTIVE: The aim of the present study was to evaluate the hemodynamic effects of acute hyperglycemia in type 2 diabetic patients and to see whether these effects are related to changes in nitric oxide (NO) availability. RESEARCH DESIGN AND METHODS: Twenty newly diagnosed complication-free diet-treated type 2 diabetic patients participated in the study. All patients underwent 3 hyperglycemic glucose clamps in random order: 1) the control study was performed with plasma glucose clamped at 18 mmol/l for 2 h; 2) the octreotide study with plasma insulin blocked at basal levels during the clamp; and 3) the L-arginine study with L-arginine (1 g/min) infused during the last 30 min of the clamp. A group of 8 patients also underwent a glutathione infusion (600 mg as an intravenous bolus followed by 5 mg/min infusion) during the clamp. RESULTS: During hyperglycemia, there were significant increments of systolic (sBP) (from 115.5 +/- 9.1 to 120.3 +/- 8.2 mmHg, P < 0.01) and diastolic (dBP) (from 70.3 +/- 7.8 to 79.7 +/- 5.3 mmHg, P < 0.01) blood pressure, as well as heart rate (from 75.2 +/- 7.8 to 80.8 +/- 5.4 beats/min, P < 0.01) and plasma catecholamines (P < 0.05). Squatting ratios, a measure of the baroreflex activity, significantly deteriorated after hyperglycemia (P < 0.01). The infusion of octreotide, used to avoid the possible confounding influence of insulin, did not change the hemodynamic effects of hyperglycemia. Glutathione, a free radical scavenger, completely prevented the vascular effects of hyperglycemia. L-Arginine produced a fall in sBP and dBP to baseline values and normalized squatting ratios. CONCLUSIONS: Acute hyperglycemia in newly diagnosed type 2 diabetic patients causes significant hemodynamic changes that are independent of endogenous insulin and are prevented by glutatione and reversed by L-arginine, suggesting an interference with endogenous NO availability. These observations could help explain the adverse cardiovascular effects of hyperglycemic spikes. PMID- 10834427 TI - Urinary transforming growth factor-beta1 and alpha1-microglobulin in children and adolescents with type 1 diabetes. AB - OBJECTIVE: Transforming growth factor (TGF)-beta1 is an important mediator in the pathogenesis of diabetic nephropathy. Urinary TGF-beta1 reflects TGF-beta1 production in the kidney, and alpha1-microglobulin tubular dysfunction. These 2 markers were studied in the early phases of type 1 diabetes. RESEARCH DESIGN AND METHODS: There were 113 type 1 diabetic children and adolescents (mean +/- SD: age 14.1 +/- 2.9 years, and diabetes duration 7.4 +/- 2.9 years, HbA1c 9.3 +/- 1.5%) and 39 healthy subjects (age 13.8 +/- 2.8 years) who participated in the study. Of the diabetic patients, 105 were normoalbuminuric (2-3 consecutive overnight urinary albumin excretion rates [AERs] <20 microg/min) and 8 had microalbuminuria (at least 2 AERs 20-200 microg/min). Overnight urinary TGF-beta1 and alpha1-microglobulin levels were measured and the results expressed as the ratio to urinary creatinine concentration. RESULTS: Data are medians (range). Diabetic patients had higher urinary TGF-beta1 levels than those of control subjects: 0.9 ng/mg (0.05-122.3) vs. 0.3 ng/mg (0.05-2.2) creatinine, respectively (P = 0.003). Urinary TGF-beta1 levels correlated with urinary glucose (r = 0.2, P = 0.03) and alpha1-microglobulin (r = 0.2, P = 0.02) levels, but not with HbA1c, AER, age, or duration of diabetes. In 43 patients with urinary TGF-beta1 above the control levels, urinary TGF-beta1 levels correlated with urinary glucose (r = 0.6, P < 0.001) and alpha1-microglobulin (r = 0.6, P < 0.001) levels. Diabetic patients had higher urinary alpha1-microglobulin levels than those of control subjects: 4.8 microg/mg (0.6-48.8) vs. 2.7 microg/mg (0.8 11.6) creatinine, respectively (P < 0.001). Alpha1-microglobulin levels correlated with AER (r = 0.2, P = 0.02), HbA1c (r = 0.3, P = 0.001), urinary glucose (r = 0.5, P < 0.001), and urinary TGF-beta1 levels. CONCLUSIONS: An early rise in urinary TGF-beta1 levels was observed in young type 1 diabetic patients. Urinary TGF-beta1 is associated with 2 interrelated tubular markers, alpha1 microglobulin and urinary glucose. PMID- 10834428 TI - Apolipoprotein E isoform polymorphisms are not associated with insulin resistance: the Framingham Offspring Study. AB - OBJECTIVE: Insulin resistance and the apolipoprotein (apo) allele e4 have both been associated with coronary heart disease (CHD). We examined the relationship between insulin resistance and apo(e) polymorphisms among participants in the Framingham Offspring Study. RESEARCH DESIGN AND METHODS: During 1991-1995, subjects underwent a clinical examination and an oral glucose tolerance test with measurement of fasting and 2-h glucose, insulin levels, and fasting lipid levels. We measured insulin resistance using the homeostasis model, in which insulin resistance (HOMA-IR) = fasting insulin x glucose/22.5. Apo(e) isoforms and phenotypes were determined in 1983-1987 using isoelectric focusing of plasma VLDL. Of the 2,120 subjects with complete HOMA-IR and apo(e) data, 204 with type 2 diabetes were excluded. The remainder were classified with features of the insulin resistance syndrome including impaired glucose tolerance (1997 American Diabetes Association criteria), hypertension (criteria from the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure [6th report]), obesity (BMI >85th percentile), high waist-to-hip ratio (>85th percentile), and high triglyceride and low HDL levels (NCEP-2 criteria). We analyzed data with contingency tables and age- and sex-adjusted logistic regression models. RESULTS: Among the 1,916 subjects, the mean age was 55 (range 28-83 years) and 51% were women. Median HOMA-IR was 6.4 (interquartile range 5.2 8.2), and allele frequencies were 7.8, 79.9, and 12.4% for apo(e) alleles e2, e3, and e4, respectively There were no differences in proportions of apo(e) isoforms or alleles across increasing quintiles of HOMA-IR. A less dramatic increase in proportions occurred with elevated triglycerides associated with increasing HOMA IR among those with apo(e) isoforms 2/2 and 2/3 compared with the others (P < or = 0.01 for interaction). Otherwise, apo(e) did not substantially modify associations between insulin resistance and features of the insulin resistance syndrome. CONCLUSIONS: There is no association between apo(e) polymorphisms and insulin resistance. These appear to represent 2 completely independent risk factors for CHD. PMID- 10834429 TI - Repaglinide acutely amplifies pulsatile insulin secretion by augmentation of burst mass with no effect on burst frequency. AB - OBJECTIVE: Repaglinide is a new oral hypoglycemic agent that acts as a prandial glucose regulator proposed for the treatment of type 2 diabetes by stimulating insulin secretion. The aim of this study was to explore actions of repaglinide on the rapid pulsatile insulin release by high-frequency insulin sampling and analysis of insulin-concentration time series. RESEARCH DESIGN AND METHODS: We examined 8 healthy lean male subjects in a single-dose double-blind placebo controlled crossover design. After the subjects underwent an overnight fast, blood sampling was initiated and continued every minute for 120 min. After 40 min, a single dose (0.5 mg) of repaglinide or placebo was given. Serum insulin concentration time series were assessed by deconvolution analyses and the regularity statistic by approximate entropy (ApEn). RESULTS: Average insulin concentration was increased after repaglinide administration (basal vs. stimulated period, P values are placebo vs. repaglinide) (25.1 +/- 3.6 vs. 33.5 +/- 4.1 pmol/l, P < 0.001). Insulin secretory burst mass (15.8 +/- 2.2 vs. 19.6 +/- 2.8 pmol x l(-1) x pulse(-1), P = 0.02) and amplitude (6.1 +/- 0.9 vs. 7.7 +/ 1.2 pmol x l(-1) x min(-1), P = 0.008) were augmented after repaglinide administration. A concomitant trend toward an increase in basal insulin secretion was observed (2.5 +/- 0.3 vs. 3.2 +/- 0.4 pmol x l(-1) x min(-1), p = 0.06), while the interpulse interval was unaltered (6.8 +/- 1.0 vs. 5.4 +/- 0.4 min/pulse, P = 0.38). ApEn increased significantly after repaglinide administration (0.623 +/- 0.045 vs. 0.670 +/- 0.034, P = 0.04), suggesting less orderly oscillatory patterns of insulin release. CONCLUSIONS: In conclusion, a single dose of repaglinide amplifies insulin secretory burst mass (and basal secretion) with no change in burst frequency. The possible importance of these mechanisms in the treatment of type 2 diabetes characterized by disrupted pulsatile insulin secretion remains to be clarified. PMID- 10834430 TI - National standards for diabetes self-management education. Task Force to Review and Revise the National Standards for Diabetes Self-Management Education Programs. PMID- 10834431 TI - American Diabetes Association annual meeting, 1999: dyslipidemia, endothelial dysfunction, and glycosylation. PMID- 10834432 TI - Venlafaxine HCl in the treatment of painful peripheral diabetic neuropathy. PMID- 10834433 TI - Bilateral diabetic infarction of the anterior tibial muscle. PMID- 10834434 TI - Function of pancreatic islets isolated from a type 1 diabetic patient. PMID- 10834435 TI - Circulating levels of coagulation factor XIII in subjects with type 2 diabetes and in their first-degree relatives. PMID- 10834436 TI - Effect of aldose reductase inhibitor on cutaneous nerve fiber length in diabetic patients. PMID- 10834437 TI - Prevalence of diabetes in adult patients with Down's syndrome living in a residential home. PMID- 10834438 TI - Effect of data management on a central server on HbA1c levels in insulin requiring patients. PMID- 10834439 TI - Prevalence of diabetes, impaired glucose tolerance, and impaired fasting glucose in a rural population of Korea, according to 1997 American Diabetes Association and 1985 World Health Organization criteria. PMID- 10834440 TI - Rapid remission of nephrotic-range proteinuria in a case of histologically proven diabetic nephropathy treated with an ACE inhibitor. PMID- 10834441 TI - Interesting insulin response to oral glucose load in young Japanese subjects with impaired glucose tolerance. PMID- 10834442 TI - An equation for insulin sensitivity index. PMID- 10834443 TI - Effect of gluten-free diet on the metabolic control of type 1 diabetes in patients with diabetes and celiac disease. PMID- 10834444 TI - Prediction of albumin excretion rate from albumin-to-creatinine ratio. PMID- 10834445 TI - Is there any use for the oral glucose tolerance test? PMID- 10834446 TI - Successful launch of cardiac transplantation in Japan. Osaka University Cardiac Transplant Program. AB - Cardiac transplantation has been established as a therapeutic strategy for patients with end-stage heart failure. In Japan, however, cardiac transplantation has not been performed since the first case in 1968, and even now, after legislation for the approval of brain death was passed in 1997, it is still not performed regularly. Following long and steady efforts to enlighten Japanese society about the concept of brain death and the importance of organ transplantation, the first cardiac transplantation under the new legislation was successfully performed at Osaka University Hospital on February 1999. The patient was 47-year-old male in the dilated phase of hypertrophic cardiomyopathy who had been supported with an implantable left ventricular assist device. This article briefly reviews the situation prior to the first case of cardiac transplantation under the new legislation and discusses the current status of the therapy in Japan. PMID- 10834448 TI - Prognostic value of left atrial appendage function in patients with dilated cardiomyopathy. AB - The purpose of the present study was to determine whether parameters of left atrial appendage (LAA) function, assessed by transesophageal echocardiography (TEE), could predict the clinical outcome in patients with dilated cardiomyopathy (DCM). Fifty-five patients (20 had ischemic cardiomyopathy; mean age, 56+/-14 years) who underwent TEE to evaluate LAA function from 1992 to 1996 were studied. After a mean follow-up period of 34+/-13 months, 16 patients died; the cause was cardiac in 14 and noncardiac in 2. Patients who died of cardiac cause had a lower LAA emptying velocity than survivors (38+/-18 vs 54+/-18 cm/s, p=0.01). There were, however, no significant differences between survivors and nonsurvivors with regard to the maximal LAA area (4.3+/-1.3 vs 4.5+/-0.9 cm2, p=0.55), minimal LAA area (2.4+/-1.1 vs 2.9+1.1 cm2, p=0.13), and LAA ejection fraction (46+/-16 vs 36+/-18%, p=0.05). On the Cox proportional hazards model analysis, LAA emptying velocity <50 cm/s (chi-square 5.9, p=0.02), LAA ejection fraction <43% (chi square 5.6, p=0.02), female gender (chi-square 5.2, p=0.02), pulmonary artery wedge pressure > or =14 mmHg (chi-square 4.8, p=0.03), E/A ratio > or =1.3 (chi square 4.6, p=0.03), deceleration time <148 ms (chi-square 4.6, p=0.03), and cardiothoracic ratio > or =54% (chi-square 4.3, p=0.04) were significantly related to cardiac death. The stepwise multivariate analysis revealed that LAA emptying velocity (chi-square 6.1, p=0.01) and gender (chi-square 5.4, p=0.02) were the independent predictors for outcome. In conclusion, the parameters of LAA function may be useful predictors of the clinical outcome in patients with DCM. PMID- 10834447 TI - Cardiac transplantation under new legislation for organ transplantation in Japan: report of two cases. AB - During the past 2 years since new legislation for organ transplantation from brain-dead donors came into effect in Japan, 3 cardiac transplants have been carried out, 2 of which were performed at the National Cardiovascular Center (NCVC). The recipient cases were 46- and 25-year-old male patients who suffered from end-stage dilated cardiomyopathy and had been listed for cardiac transplantation in the Japan Organ Transplantation Network as status I candidates. The first patient was supported by the use of a paracorporeal air driven left ventricular assist device of the NCVC type, and had a moderate degree of renal and hepatic dysfunction at the time of transplantation. Donor hearts were transported from distant hospitals (Tokyo and Miyagi prefecture) and the transportation time was 1 h 33 min and 2h 4 min, respectively. The operation was performed by the standard technique (Lower-Shumway) in the first patient and by the bicaval anastomosis technique in the second patient. Reperfusion of the transplanted heart was performed retrogradely through the coronary sinus utilizing leukocyte-depleted blood with a gradual increase in temperature. Total ischemic time was 3 h 34 min and 3 h 35 min, respectively. Weaning from the cardiopulmonary bypass was easy and uneventful in each patient. Immunosuppressive therapy was conducted with OKT-3 induction in the first patient because of the coexisting renal dysfunction and with a triple immunosuppressive regimen for both patients. Routine endomyocardial biopsy showed acute rejection of less than grade Ib, and the patients were discharged on the 65th and 46th postoperative day, respectively. At present, both patients are in the NYHA class I state and are ready to return to work. The uneventful recovery seen in these patients shows the advances made in transplant medicine, including the progress and improvement of immunosuppressive therapy, surgical techniques, myocardial protection, and detection and treatment of infection. Further efforts are required to fully establish the cardiac transplantation program in Japan. PMID- 10834449 TI - Reverse redistribution of thallium-201 myocardial single photon emission tomography and contractile reserve. AB - The present study investigated the contractile reserve of myocardium exhibiting reverse redistribution (RRD) of thallium-201 (201Tl) after acute myocardial infarction. Forty patients experiencing their first acute myocardial infarction underwent resting 201Tl single-photon emission computed tomography (SPECT) and low-dose (5-10 microgxkg(-1)xmin(-1)) dobutamine stress echocardiography (DSE) within 4 weeks after the onset of infarction. The left ventricle was divided into 13 segments for analysis. The severity of defects in 201Tl SPECT and the extent of wall motion abnormality in DSE were visually assessed and scored. The sum of each defect score and wall motion score of infarct-related segments were defined as total defect score (TDS) and total wall motion score (TWM), respectively. Quantitative analysis of 201Tl uptake was also performed. Resting 201Tl SPECT revealed RRD in 16 patients (group RRD), fixed defect (FIX) in 23 patients (group FIX), and redistribution in one. There was a significant difference in improvement of TWM between rest and stress in TWM in both the RRD and FIX groups (p<0.0001, each case). The improvement of TWM with dobutamine was significantly greater in RRD than in FIX (1.6+/-1.0 vs 0.6+/-0.7, p=0.001). There was a positive correlation between the magnitude of RRD and improvement of TWM with dobutamine (r=0.48, p=0.002). Myocardium exhibiting RRD on 201Tl SPECT in patients with acute myocardial infarction has greater contractile reserve than that exhibiting a fixed defect. PMID- 10834450 TI - Association of matrix metalloproteinase expression and left ventricular function in idiopathic dilated cardiomyopathy. AB - Myocardial remodeling is an important predictor for the development of dilated cardiomyopathy (DCM). Matrix metalloproteinases (MMPs) are the family of proteins responsible for extracellular remodeling, and tissue inhibitors of metalloproteinases (TIMPs) tightly control their activity. In the present study, the expression of MMP-2, MMP-9, TIMP-1 and TIMP-2 was determined by immunohistochemistry in right ventricular endomyocardial biopsy samples from 16 patients with idiopathic DCM, and its clinical significance was evaluated by comparison with parameters of cardiac function. To obtain a semi-quantitative assessment of MMP and TIMP expression, the average number of positive cells per high power field was counted. The left ventricular ejection fraction (LVEF) significantly correlated with the expression of both MMP-2 (r=-0.68) and TIMP-2 (r=-0.58). Patients were classified into 2 groups according to the degree of MMP 2 expression: strongly positive and weakly positive. LVEF, left ventricular (LV) end-diastolic pressure, right ventricular end-diastolic pressure, pulmonary capillary wedge pressure and the plasma norepinephrine level were significantly greater in the strongly positive group (p<0.05). In conclusion, the expression of MMPs and TIMPs in the cardiac matrix of patients with idiopathic DCM is closely associated with myocardial remodeling and subsequent deterioration of LV performance. These findings suggest new therapeutic targets for patients with idiopathic DCM. PMID- 10834451 TI - Retrospective analysis showing less cardiac events in post-myocardial infarction patients treated with metoprolol. Secondary Prevention Group. AB - This analysis was carried out to clarify the capacity of metoprolol to prevent cardiac events in Japanese post-myocardial infarction patients during a follow-up period of 16.3 months. Cardiac events occurred in 44 of 650 patients treated without beta-blockers (6.8%) and in 13 of 432 patients treated with metoprolol (3.0%), which represents a significant decline in the incidence of cardiac events among patients receiving metoprolol (p<0.01, odds ratio 0.43, 95% confidence interval 0.23-0.80). Because this was a retrospective analysis, there were unavoidable differences in the backgrounds of the patients in the 2 groups. Subgroup analyses, each focusing on a specific patient characteristic, were therefore performed. These showed that metoprolol effectively reduced cardiac events in many subgroups. Furthermore, multivariate analysis carried out to exclude any modification based on the differences in patient background confirmed metoprolol to be effective in reducing subsequent cardiac events in post myocardial infarction patients. A large, randomized, placebo-controlled clinical trial needs to be performed in the Japanese population to confirm the present result. PMID- 10834452 TI - Effect of beta-blocker on left ventricular function and natriuretic peptides in patients with chronic heart failure treated with angiotensin-converting enzyme inhibitor. AB - To evaluate whether or not beta-blockers can improve the condition of patients with heart failure treated with a combination of diuretics, digitalis and angiotensin-converting enzyme inhibitor (ACEI), 52 patients with chronic heart failure who have been treated with ACEI for more than 6 months were enrolled. They were divided into 2 groups: 26 patients continued the same therapy another 6 months or more (group A), and 26 patients were given oral metoprolol for 6 months or more, in addition to the ACEI (group B). Echocardiographic parameters and atrial and brain natriuretic peptides (ANP, BNP) were measured. The left ventricular dimensions at end-diastole and end-systole were significantly decreased and fractional shortening was significantly increased in group B after 6 months' treatment with the beta-blocker, but these parameters remained unchanged in group A. Plasma levels of both ANP and BNP were significantly decreased in group B, but remained unchanged in group A. These results indicate that concomitant beta-blocker therapy can improve left ventricular function and attenuate plasma ANP and BNP levels in patients with chronic heart failure treated with ACEI. PMID- 10834453 TI - Hemodynamic and thermal responses to a 30-minute constant-workload aerobic exercise in middle- or old-aged patients with cardiovascular diseases. AB - The aim of the present investigation was to compare the hemodynamic and thermal responses to a 30-min aerobic exercise between middle- or old-aged patients with normal left ventricular function and those with left ventricular dysfunction. Constant-load sitting ergometer exercise of approximately 90% of the subject's oxygen uptake (VO2) at the anaerobic threshold for 30 min was conducted in 21 patients with left ventricular dysfunction (61+/-10 years, left ventricular ejection fraction (LVEF) 35+/-7%) and 24 patients with normal left ventricular function (59+/-9 years, LVEF 71+/-7%). Heart rate (HR), blood pressure, deep temperatures in the forehead and thigh, and forearm skin blood flow (SkBF) were measured every minute, and cardiac output (CO) and stroke volume (SV) were determined every 10 min with the dye-dilution technique during the exercise. Patients of both groups exhibited a progressive elevation in each temperature and an increase in SkBF during the exercise. Although the VO2 and CO remained stable, almost the same magnitude of decrease in SV as increase in HR was seen after the 10th min of exercise in both groups. The magnitude of the decrease in SV was greater in old-aged than middle-aged patients with left ventricular dysfunction. Thus, the downward drift in SV during a 30-min constant-load aerobic exercise might not be influenced by left ventricular function, but intensified by aging in patients with left ventricular dysfunction. PMID- 10834454 TI - Magnesium dynamics and relation to left ventricular function in acute myocardial infarction. AB - The present study investigated the serial changes in serum magnesium (Mg) and erythrocyte concentration of Mg in patients with acute myocardial infarction (AMI) and the relationship between these changes and left ventricular ejection fraction (LVEF) at 1 month after the onset of infarction. The study group comprised 26 patients with AMI (mean age, 57.9+/-8.9 years). Serum Mg and erythrocyte Mg were measured on hospital days 1, 2, 4, 7 and 21. The change in erythrocyte Mg during the acute phase was calculated as a ratio: [(erythrocyte Mg at day 2)-(erythrocyte Mg at day 1)]/(erythrocyte Mg at day 1). The change in serum Mg was calculated similarly. The following results were obtained. (1) Serum Mg tended to increase from the onset of myocardial infarction (day 1: 1.86+/ 0.19, day 2: 1.93+/-0.22, day 4: 2.17+/-0.23; day 7: 2.25+/-0.20; day 21: 2.12+/ 0.15 mg/dl). (2) Erythrocyte Mg on day 2 and day 4 showed a significant decrease compared with day 1 (day 1: 2.45+/-0.40, day 2: 2.09+/-0.41, day 4: 2.07+/-0.37, day 7: 2.22+/-0.33, day 7: 2.34+/-0.28 mg/dl per 400x10(4)/mm3 cells). (3) A significant positive correlation was observed between the change in serum Mg and LVEF (r=0.55, p<0.05), and a significant negative correlation was observed between the change in erythrocyte Mg and LVEF (r=-0.57, p<0.05). Thus, it was concluded that an extracellular shift in intracellular Mg occurred during the first 2 days after the onset of myocardial infarction. This responsive increase in the extracellular Mg level may be an important factor for maintaining left ventricular function in patients 1 month after the onset of AMI. PMID- 10834455 TI - Congestive heart failure after peripheral blood stem cell transplantation: role of cytokines. AB - A 32-year-old woman was admitted with the diagnosis of congestive heart failure (CHF) without organic heart disease after peripheral blood stem cell transplantation (PBSCT) for malignant lymphoma. Various cytokines have been reported to be released from stem cells after PBSCT and some have a suppressive effect on myocardial contractility; elevated levels of cytokines have been reported in dilated cardiomyopathy (DCM) and/or CHF patients. In the present case, elevated levels of interleukin (IL)-6 and tumor necrosis factor-alpha (TNF alpha) were observed, and there was a parallel relationship between the recovery of cardiac function and the decrease of these cytokines, strongly suggesting that the release of IL-6 and TNF-alpha after PBSCT might have been important in the pathogenesis of the CHF. PMID- 10834456 TI - Primary pericardial mesothelioma presenting as constrictive pericarditis: a case report. AB - Primary malignant pericardial mesothelioma is a rare tumor and the case reported here presented as constrictive pericarditis. The patient's symptoms progressed day by day despite treatment with digitalis, diuretics and catecholamines. Although a computed tomographic scan of the chest, echocardiography and pericardiocentesis were performed, a preoperative definitive diagnosis could not be obtained. Emergency pericardiectomy and partial resection of the tumor were carried out with the aid of a percutaneous cardiopulmonary supporting system, but the patient died of cardiac failure on postoperative day 3. The tumor appeared to be the biphasic type of diffuse malignant mesothelioma. The prognosis for pericardial mesothelioma is extremely poor due to its late presentation and difficulty in completely removing it surgically and, unfortunately, there still is not a radical therapy for this tumor. PMID- 10834457 TI - Subacute tuberculous pericarditis with fibroelastic constriction diagnosed upon pericardiectomy. AB - A patient with subacute pericarditis showed no evidence suggesting tuberculosis until pericardiectomy was performed because of hemodynamic deterioration. The excised pericardium had a rubbery fibroelastic consistency; histologically, there were granulomatous changes characteristic of tuberculosis. Although tuberculous pericarditis is a difficult diagnosis, this case illustrates the diagnostic and therapeutic importance of early pericardiectomy before myocardial inflammatory infiltration occurs together with end-stage pericardial fibrosis and calcification. PMID- 10834458 TI - Radiofrequency catheter ablation of a concealed atrioventricular accessory pathway associated with a coronary sinus diverticulum. AB - A 31-year-old woman underwent radiofrequency catheter ablation of a concealed left posteroseptal accessory pathway associated with a coronary sinus diverticulum. The patient had previously undergone unsuccessful catheter ablation of the posteroseptal region of the mitral annulus. Coronary sinus venography revealed the presence of the diverticulum near the ostium. An electrogram in the neck of the diverticulum showed the shortest ventriculoatrial conduction time and a large accessory pathway potential during atrioventricular reciprocating tachycardia. The pathway was successfully ablated within the neck of the diverticulum. The findings in this case underscore the importance of coronary sinus venography before ablation. PMID- 10834459 TI - Refractory coronary spasm relieved by intracoronary administration of nicorandil. AB - Two patients in whom coronary spasm was refractory to intracoronary injection of nitroglycerin were relieved by intracoronary administration of nicorandil (a nitrate and potassium channel opener) during catheterization. These findings suggest that nicorandil may prove useful as an additional therapeutic agent. PMID- 10834460 TI - Transient complete atrioventricular block during catheter ablation of left free wall bypass tract. AB - Radiofrequency catheter ablation of accessory bypass tracts associated with the Wolff-Parkinson-White (WPW) syndrome has become an accepted and widespread therapy. When bypass tracts are located in the free wall of the left ventricle, complete atrioventricular (AV) block is an unusual complication. Two cases of symptomatic WPW syndrome with transient complete atrioventricular block during catheter ablation are described. The first case was a 14-year-old female with an accessory pathway located in the left posterior wall, and the second was a 72 year-old female with an accessory pathway located in the left lateral wall. Radiofrequency energy application resulted in transient complete AV block with escape rhythm. In the first case, AV conduction with left bundle branch block resumed the next day, whereas in the second case, AV conduction soon resumed with prolongation of atrio-His (AH) interval and no evidence of pre-excitation. This phenomenon could have been due to either trauma to the AV node during catheter entry into the left ventricle or compression of the AV node with a catheter shaft during ablation because both patients' hearts were comparatively small. PMID- 10834461 TI - Coronary artery bypass grafting following substernal gastric interposition. AB - A 61-year-old man who had undergone substernal esophagogastric anastomosis for reconstruction after esophageal cancer, developed unstable angina 9 years later. Complete revascularization for triple-vessel disease was performed via a left thoracotomy approach under cardiopulmonary bypass. The successful results show that complete revascularization can be performed via this approach. PMID- 10834462 TI - Successful treatment of malignant neurocardiogenic syncope with repeated tilt training program. AB - Recent reports have shown that repeated tilt-table testing or tilt training is a very effective therapy for the treatment of neurocardiogenic syncope induced by head-up tilt testing. The present patient experienced repeated syncopal or presyncopal attacks and had shown prolonged asystole on an electrocardiogram during syncope. The presyncope could be reproducibly induced by head-up tilt testing. Oral propranolol and/or disopyramide therapies failed to prevent his symptoms. Tilt training (2 sessions/day) was repeated every day for 4 weeks at home, and then head-up tilt testing was performed again. The syncope or presyncope was not induced by head-up tilt testing. The patient has continued with this training and has had no symptoms during the follow-up period of 1 year. PMID- 10834463 TI - Physiological self-regulation and information processing in infancy: cardiac vagal tone and habituation. AB - This study investigates the role of physiological self-regulation (cardiac vagal tone) in information processing (habituation) in 81 infants. Nucleus ambiguus vagal tone (Vna, a measure of respiratory sinus arrhythmia) was used to index cardiac vagal tone. Physiological self-regulation was operationalized as the change in Vna from a baseline period of measurement to habituation. Decreases in Vna consistently related to habituation efficiency, operationalized as accumulated looking time (ALT), in all infants twice at 2 months and twice at 5 months; however, this relation was accounted for by infants who met an habituation criterion on each task. Among habituators, shorter lookers also had greater Vna suppression during habituation. Within-age and between-age suppression of vagal tone predicted ALT, but ALT did not predict suppression of vagal tone. Physiological self-regulation provided by the vagal system appears to play a role in information processing in infancy as indexed by habituation. PMID- 10834464 TI - Compulsive-like behavior in individuals with Down syndrome: its relation to mental age level, adaptive and maladaptive behavior. AB - This study examined the nature of repetitive, ritualistic, and compulsive-like behaviors in 50 typically developing children and 50 individuals with Down syndrome (DS), matched on mental age (MA; M = 59.72 months). Parents reported on their children's compulsive-like behaviors-including ritualistic habits-and perfectionistic behaviors, as well as their children's adaptive and maladaptive behaviors. Results indicated that children with DS show similar MA-related changes in compulsive-like behaviors compared to the MA-matched comparison group. Younger children (both typical and DS) exhibited significantly more compulsive like behaviors than older children. In general, children with and without DS did not differ from each other in terms of the number of compulsive-like behaviors they engaged in, although participants with DS engaged in more frequent, more intense repetitive behaviors. Compulsive-like behaviors were differentially related to adaptive and maladaptive behaviors across the MA and mental retardation groups. The results extend the "similar sequence" model of development to the construct of compulsive-like behaviors, and also suggest that some repetitive behaviors may be among the behavioral phenotype of individuals with DS. PMID- 10834465 TI - Crying and infant abuse in rhesus monkeys. AB - This study investigated the relation between crying and infant abuse in group living rhesus monkeys (Macaca mulatta). The subjects were 10 abusive mothers with their infants and 10 control mother-infant pairs. Abused infants cried more frequently than controls in the first 12 weeks of life, even when cries immediately following abuse were excluded from the analysis. The coos of 5 abused infants differed from those of 5 controls in several acoustic parameters, whereas their screams and geckers were acoustically similar, when recorded in the same context. Abusive mothers were less likely than control mothers to respond positively to the cries of their infants. Although infant cries may increase the probability of abuse being repeated, infant crying per se does not appear to be a major determinant of abuse. PMID- 10834466 TI - Measurement properties of the MacArthur communicative development inventories at ages one and two years. AB - In a prospective study of child development in relation to early-life otitis media, we administered the MacArthur Communicative Development Inventories (CDI) to a large (N = 2,156), sociodemographically diverse sample of 1- and 2-year-old children. As a prerequisite for interpreting the CDI scores, we studied selected measurement properties of the inventories. Scores on the CDI/Words and Gestures (CDI-WG), designed for children 8 to 16 months old, and on the CDI/Words and Sentences (CDI-WS), designed for children 16 to 30 months old, increased significantly with months of age. On several scales of both CDI-WG and CDI-WS, standard deviations approximated or exceeded mean values, reflecting wide variability in results. Statistically significant differences in mean scores were found according to race, maternal education, and health insurance status as an indirect measure of income, but the directionality of differences was not consistent across inventories or across scales of the CDI-WS. Correlations between CDI-WG and CDI-WS ranged from .18 to .39. Our findings suggest that the CDI reflects the progress of language development within the age range 10 to 27 months. However, researchers and clinicians should exercise caution in using results of the CDI to identify individual children at risk for language deficits, to compare groups of children with different sociodemographic profiles, or to evaluate the effects of interventions. PMID- 10834467 TI - Measuring variability in early child language: don't shoot the messenger. AB - Feldman et al. criticize the MacArthur Communicative Development Inventories (CDIs) as having too much variability, too little stability, and insufficient ability to predict early language delay. We present data showing that these characteristics of the CDI are authentic reflections of individual differences in early language development rather than measurement deficiencies. We also respond to their critical assertions concerning sociodemographic influences on the CDI scores. PMID- 10834468 TI - Dual representation and young children's use of scale models. AB - To use a symbolic object such as a model, map, or picture, one must achieve dual representation; that is, one must mentally represent both the symbol itself and its relation to its referent. The studies reported here confirm predictions derived from this concept. As hypothesized, dual representation was as difficult for 2 1/2-year-olds to achieve with a set of individual objects as it was with an integrated model. Decreasing the physical salience of a scale model (by placing it behind a window) made it easier for 2 1/2-year-old children to treat it as a representation of something other than itself. Conversely, increasing the model's salience as an object (by allowing 3-year-old children to manipulate it) made it more difficult to appreciate its symbolic import. The results provide strong support for dual representation. PMID- 10834469 TI - Relating quality of center-based child care to early cognitive and language development longitudinally. AB - How quality of center-based child care relates to early cognitive and language development was examined longitudinally from 6 to 36 months of age in a sample of 89 African American children. Both structural and process measures of quality of child care were collected through observation of the infant classroom. Results indicated that higher quality child care was related to higher measures of cognitive development (Bayley Scales of Infant Development), language development (Sequenced Inventory of Communication Development), and communication skills (Communication and Symbolic Behavior Scales) across time, even after adjusting for selected child and family characteristics. In addition, classrooms that met professional recommendations regarding child:adult ratios tended to have children with better language skills. Classrooms that met recommendations regarding teacher education tended to have girls with better cognitive and receptive language skills. These findings, in conjunction with the growing child-care literature, provide further evidence that researchers and policymakers should strive to improve the quality of child care to enhance early development of such vulnerable children. PMID- 10834470 TI - Early maternal and child influences on children's later independent cognitive and social functioning. AB - The present study examined whether parenting and child characteristics of 2- and 3 1/2-year-old children had common paths of influence on their 4 1/2-year independent cognitive and social functioning. Structural equation modeling was guided by hypotheses that assumed children's later independence is facilitated by specialized parental support in early social interactions. To address the importance of variability in early development for understanding children's later independence, we included 104 term and 185 preterm children, as they are known to differ in early skills. As predicted, mothers' maintaining of children's interests indirectly supported 4 1/2-year cognitive and social independence through a direct, positive influence on 2- and 3 1/2-year skills. Directiveness positively supported children's early cognitive and responsiveness skills but by 3 1/2 years, high levels of this behavior had a direct, negative influence on their cognitive and social independence at 4 1/2 years. Whereas high levels of maintaining interests across these ages support later independence, directiveness needs to decrease in relation to children's increasing competencies. Results support a theoretical framework that emphasizes the importance of the social context for understanding the origins of children's later independent functioning. PMID- 10834471 TI - The effects of global severe privation on cognitive competence: extension and longitudinal follow-up. English and Romanian Adoptees Study Team. AB - The current study extends previous research on a sample of children adopted into the United Kingdom following severe early deprivation and a comparison sample of nondeprived, within-country, early adoptees. We assessed 165 children adopted from Romania and 52 U.K. adoptees at age 6 years. Longitudinal data (at age 4 and 6 years) were available on 111 Romanian adoptees placed into U.K. homes before 24 months of age and on all U.K. adoptees. Results indicated that there was considerable catch-up among late-placed Romanian children from entry into the United Kingdom to age 6, but as a group they exhibited lower cognitive scores and general developmental impairment compared with earlier adopted Romanian children. In addition, the resilience suggested at the assessment at age 4 years was maintained longitudinally, but there was no further evidence of catch-up or recovery. PMID- 10834472 TI - Development of face processing: the effect of face inversion. AB - The present experiment examined the degree to which analytic and holistic modes of processing play a role in the way children and adults categorize upright and inverted faces. Seven-year-old children (n = 38), 10-year-old children (n = 40), and adults (n = 55) were instructed to classify upright and inverted faces into two categories. The construction of the categories allowed participants to categorize the faces either analytically (by focusing on a single attribute) or holistically (in terms of overall similarity). The results show both a developmental trend from analytic to holistic processing and an effect of face inversion with increasing age. Thus, it appears that 7-year-old children process upright and inverted faces in a way comparable to their processing of nonfacial visual stimuli, namely analytically, whereas a growing proportion of 10-year-olds and adults process only upright faces holistically by adopting a specific mode of face processing. PMID- 10834473 TI - A tale of two representations: the misinformation effect and children's developing theory of mind. AB - The present research investigates representational ability as a cognitive factor underlying the suggestibility of children's eyewitness memory. The misinformation effect is used as an index of children's suggestibility, and performance on the false belief task is used as an assessment of children's representational abilities (N = 117). Analyses that considered the effect of representational ability and general memory ability on children's susceptibility to misleading information showed that differences in representational ability and general memory ability predicted participants' susceptibility to misleading information. These results demonstrate that the eyewitness memory of children who lack either multirepresentational abilities, sufficient general memory abilities, or both (i.e., most 3- and 4-year-olds) is less accurate than the eyewitness memory of children with both multirepresentational abilities and sufficient memory abilities (i.e., most 6-year-olds and adults). Thus, it appears that the earliest age at which children's eyewitness memory can be considered to be similar to that of adults is 6 years of age, when children's mental representational abilities are similar to those of adults. These results suggest that one factor underlying children's vulnerability to misleading information is the number of representations of an event that they can simultaneously hold and compare. PMID- 10834474 TI - Mother-child mutually responsive orientation and conscience development: from toddler to early school age. AB - We examined whether positive implications of mother-child mutually responsive orientation, demonstrated earlier at toddler and preschool age, extend longitudinally into early school age. The focus of the present study was on the long-term consequences of mutually responsive orientation for the development of conscience. Mutually responsive orientation encompassed shared cooperation and shared positive affect between mother and child. It was measured as a composite of those qualities observed in dyadic naturalistic interactions and reported by mothers, at toddler and preschool age. Children's conscience was assessed at early school age (N = 83) using multiple measures, including observations of moral behavior, alone and in the peer context, and moral cognition. Mother-child mutually responsive orientation at toddler and preschool ages predicted children's future conscience, even after controlling for the developmental continuity of conscience. Model-fitting analyses revealed that mutually responsive orientation at toddler age had a direct effect on future conscience, not mediated by such orientation at preschool age. The findings extend those of earlier work that revealed the importance of mother-child mutually responsive orientation for socialization, and they confirm the value of the relationship approach to social development, including long-term outcomes. PMID- 10834475 TI - Attention-deficit/hyperactivity disordered and control boys' responses to social success and failure. AB - The behavioral, self-evaluative, and attributional responses of 120 boys with Attention-Deficit/Hyperactivity Disorder (ADHD) and 65 control boys to social success and failure were examined using a dyadic, laboratory get-acquainted task employing child confederates. Objective coders rated boys with ADHD as less socially effective than controls in their interactions, but also as less frustrated and helpless. In terms of self-evaluations, ADHD boys overwhelmingly rated their own performance more favorably than did controls and in some instances, these differences were more apparent following failure. The attributional pattern of ADHD and control boys differed in that ADHD boys were more likely than controls to attribute success to external, uncontrollable factors such as task ease and being lucky; controls, on the other hand, were more likely than ADHD boys to attribute initial failure to not having tried hard enough. Results are discussed in the context of existing literature documenting a positive illusory bias in ADHD boys' self-perceptions. PMID- 10834476 TI - Loneliness and friendship in high-functioning children with autism. AB - Loneliness and friendship were examined in 22 high-functioning children with autism and 19 typically developing children equated with the autistic children for IQ, CA, gender, mother's education, and ethnicity. Children between the ages of 8 and 14 were asked to report on both their understanding and feelings of loneliness and the quality of their friendship. Compared to typically developing children, children with autism were both lonelier and had less complete understandings of loneliness. Although all children with autism reported having at least one friend, the quality of their friendships was poorer in terms of companionship, security, and help. Fewer associations were found between loneliness and friendship for the autistic than for the non-autistic children, suggesting less understanding of the relation between loneliness and friendship. Implications of these results are discussed for conceptualizing the social deficits in autism. PMID- 10834477 TI - Toddlers' use of force against familiar peers: a precursor of serious aggression? AB - Possible precursors of serious aggression were identified in toddlers' use of force against peers. Instances of grabbing objects and hitting peers were recorded in a sample of 66 British 18- to 30-month-olds, observed at home with familiar peers and seen again 6 months later. Mothers rated aggressiveness in the context of other personality traits. Girls and boys did not differ in average levels of aggression, nor were they rated differently by the mothers. However, the observed rate of hitting peers and mothers' ratings of aggressiveness were stable over 6 months for girls, but not for boys. Toddlers who were especially sensitive to peers' possible intentions hit their peers more often. They were also more likely to use force proactively, 6 months later. PMID- 10834478 TI - Parenting and child behavioral adjustment in early childhood: a quantitative genetic approach to studying family processes. AB - The aim of this study was to examine environmental and gene-environment processes linking parenting (i.e., affect, control, responsiveness) and preschool children's behavioral adjustment difficulties (e.g., noncompliance, conduct problems) by using bivariate genetic analyses of parents' and observers' ratings. The sample included 120 identical and same-sex fraternal twin pairs (M age = 43 months). Data sources included in-home observations, interviews, and parents' reports. Observers' ratings of children's difficult behaviors included shared and nonshared environmental variance. In contrast, parents' ratings of children's conduct problems showed genetic and nonshared environmental variance. Observer rated maternal behavior included shared and nonshared environmental variance, although maternal responsiveness also included child genetic variance. Parent self-reported negative and positive affect included shared and nonshared environment as well as child genetic variance. There was no evidence for gene environment interaction or dominance. Higher levels of difficult behavior and conduct problems covaried with higher levels of maternal negative affect and control and lower levels of maternal positive affect and control. Shared environmental mediation of these correlations was found for observations, whereas genetic and nonshared environmental mediation was found for parents' ratings. In general, estimates of shared environmental variance and mediation were greatest for observational data, and estimates of child genetic variance and mediation were greatest for parent-rated data. The implications of this pattern of findings for genetic research on family processes are discussed. PMID- 10834479 TI - Psychosocial antecedents of variation in girls' pubertal timing: maternal depression, stepfather presence, and marital and family stress. AB - Drawing on Belsky, Steinberg, and Draper's evolutionary theory of the development of reproductive strategies, we tested a model of individual differences in girls' pubertal timing. This model posits that a history of psychopathology in mothers results in earlier pubertal maturation in daughters, and that this effect is mediated by discordant family relationships and father absence/ stepfather presence. The model was supported in a short-term longitudinal study of 87 adolescent girls. In the primary test of the model, it was found that a history of mood disorders in mothers predicted earlier pubertal timing in daughters, and this relation was fully mediated by dyadic stress and biological father absence. In families in which the mother's romantic partner was not the biological father, dyadic stress accounted for almost half of the variation in daughters' pubertal timing. Stepfather presence, rather than biological father absence, best accounted for earlier pubertal maturation in girls living apart from their biological fathers. We propose that stepfather presence and stressful family relationships constitute separate paths to early pubertal maturation in girls. PMID- 10834480 TI - School extracurricular activity participation as a moderator in the development of antisocial patterns. AB - This research involves a longitudinal study of antecedents and moderators in the development of antisocial patterns. Participants included 695 boys and girls who were interviewed annually from childhood to the end of high school and again at ages 20 and 24. Cluster analyses identified four configurations of boys and girls that were reasonably homogeneous with respect to behavior and academic performance at the beginning of the investigation. When tracked over time, the configurations differed significantly in patterns of early school dropout and criminal arrests. Boys and girls in the "multiple risk configuration" were more likely than those in other configurations to show long-term antisocial patterns. Participation in school extracurricular activities was associated with reduced rates of early dropout and criminal arrest among high-risk boys and girls. The decline in antisocial patterns was dependent on whether the individuals' social network also participated in school extracurricular activities. PMID- 10834481 TI - Gender effects in children's beliefs about school performance: a cross-cultural study. AB - Do young boys and girls understand what leads to academic success (e.g., talent, effort, good teaching, luck) in the same way? Do young girls and boys have equivalent perceptions of their academic competence? Are these beliefs engendered in the same way across sociocultural contexts? In a cross-cultural study of over 3,000 children in grades 2 to 6, ages 7.2 to 13.6, we discovered that boys and girls around the world have very similar ideas about what generally leads to academic success. Moreover, in the few contexts where boys' and girls' academic performances were equal, their beliefs were also equal. However, when girls outperformed boys, their beliefs in their own talent were no greater than boys' beliefs, even though they did have stronger beliefs than boys in other facets of their achievement potential (e.g., putting forth effort, being lucky, getting their teacher's help). Our findings support the generally close correspondence between children's achievement and their competence-related beliefs, with the exception that young girls appear to specifically discount their talent. The effects held regardless of the children's achievement, intelligence, or age (approximately 8 to 13 years). Girls were more biased in some contexts than in others, however, suggesting that competence-related biases are rooted in culture specific aspects of school settings. PMID- 10834482 TI - Cultural values and intergenerational value discrepancies in immigrant and non immigrant families. AB - The goal of this research was to explore the generality of developmental processes related to intergenerational value discrepancies across 701 families from immigrant and non-immigrant groups. In a study involving 471 immigrant families (197 Armenian, 103 Vietnamese, and 171 Mexican) and 230 non-immigrant families (95 African American and 135 European American), adolescents and parents reported their endorsement of values pertaining to family obligations. We examined similarities and differences at three levels of analysis, from the general to the group-specific. Results provide evidence for general developmental processes (family obligations were endorsed more by parents than by adolescents in all groups), processes associated with immigration (the intergenerational value discrepancy generally increased with time in the United States), and processes that are unique to each ethnic group. PMID- 10834483 TI - Embolization of the internal iliac artery: still more to learn. PMID- 10834484 TI - Endovascular treatment of cerebral vasospasm. PMID- 10834485 TI - Internal iliac artery embolization in the stent-graft treatment of aortoiliac aneurysms: analysis of outcomes and complications. AB - PURPOSE: To analyze the complications of internal iliac artery (IIA) embolization in conjunction with stent-graft treatment of aortoiliac aneurysms. MATERIALS AND METHODS: Seventy-one patients with aortoiliac (n = 47) or iliac (n = 24) aneurysms were treated with endoluminal placement of stent-grafts. Thirty-two patients (31 men, one woman; mean age, 73 years; range, 56-88 years) had embolization or occlusion of one (n = 27) or both (n = 5) IIAs. Status of the IIAs and the collateral circulation was assessed by retrospective review of angiographic images. Follow-up consisted of a standardized patient questionnaire and review of radiologic and medical records. RESULTS: The mean follow-up time was 35 months (range, 5-64 months). Eleven of the 47 patients with abdominal aortic aneurysms (AAA) (23%) and 19 of the 24 patients with iliac aneurysms (79%) required IIA embolization. One patient with AAA and another with iliac aneurysm had unintentional occlusion of an IIA by extension of the stent-graft over their origins. A total of seven patients had bilateral occlusion of the IIAs after the procedure. Additionally, the inferior mesenteric arteries (IMAs) of two other patients with AAA were also embolized. In six patients, all three vessels were occluded after placement of the stent-grafts. Symptoms were reported in nine of the 20 (45%) patients with iliac aneurysms and in three of the 12 (25%) patients with AAA. Symptoms consisted of buttock claudication (nine of 32, 28%), new sexual dysfunction (two of 16, 12%), and transient urinary retention (3%). Seven of the claudicants had resolution of symptoms after a mean interval of 14 months (range, 1-36 months). There were no instances of bowel ischemia, neurologic sequelae, or buttock necrosis related to these procedures. CONCLUSION: Embolization of the IIA is associated with symptoms in a significant number of patients. While symptoms are transient in most patients, they can be problematic. Efforts should be made to preserve the pelvic circulation if possible. PMID- 10834486 TI - Clinical outcome of internal iliac artery occlusions during endovascular treatment of aortoiliac aneurysmal diseases. AB - PURPOSE: To determine the clinical outcome of hypogastric artery occlusion in patients who underwent endovascular treatment of aortoiliac aneurysmal disease. MATERIAL AND METHODS: From January 1994 to March 1998, 94 patients underwent endovascular treatment of aneurysmal diseases involving the infra-abdominal aorta or iliac arteries. Preoperative and intraoperative radiologic data were reviewed. Discharge summaries, clinic visits, and phone calls formed the basis for clinical follow-up, with a mean follow-up period of 7.3 months (range, 1-24 months). RESULTS: Because of the anatomy of the aneurysms, 28 patients required occlusion of one or more hypogastric arteries. One of the 28 patients died of unrelated causes before follow-up. Seven (26%) of the remaining 27 patients developed symptoms attributable to the hypogastric artery occlusions. Five patients developed new buttock or thigh claudication; of these five patients, three with initially mild symptoms noted complete or near complete resolution of symptoms upon follow-up. One patient with originally significant claudication at 2-year follow-up noted near resolution of symptoms. The other patient with severe pain did not improve significantly on final 1-year follow-up before his death (of unrelated causes). Other clinical complications were worsening sexual function in one patient and a nonhealing sacral decubitus ulcer that developed in a debilitated patient in the postoperative setting, which required surgery. No bowel ischemia was observed. CONCLUSION: When treating aortoiliac aneurysmal disease through an endovascular approach, the occlusion of internal iliac artery is often necessary but carries with it a small but finite chance of morbidity. PMID- 10834487 TI - Hypogastric artery coil embolization prior to endoluminal repair of aneurysms and fistulas: buttock claudication, a recognized but possibly preventable complication. AB - PURPOSE: Hypogastric artery embolization is considered to be necessary to prevent retrograde flow and potential endoleaks when a stent-graft crosses the origin of the hypogastric artery. The authors assess the incidence of buttock claudication, which is the primary complication encountered. The effect of coil location and the presence of antegrade flow at the completion of embolization are evaluated. MATERIALS AND METHODS: Hypogastric artery embolization and endoluminal repair of aneurysms and fistulas was performed in 34 patients (30 men; four women) aged 27 91 years (mean, 76 years). Ten patients were being treated for solitary abdominal aortic aneurysms, 13 were being treated for aortoiliac aneurysms, and six patients were being treated for isolated common iliac aneurysms, three for hypogastric artery aneurysms and two for iliac arteriovenous fistulas. Eleven patients had coils placed completely above the bifurcation of the hypogastric artery and 23 patients had coils placed at the bifurcation, or within the branches of the hypogastric artery. Preservation of antegrade flow after embolization was noted in 14 of 34 patients. RESULTS: Thirty-four patients underwent stent-graft repair after hypogastric artery embolization. There were two perioperative deaths, three proximal leaks, and one collateral leak. Of the 32 patients who survived the procedure, there was one retrograde leak, even though 13 of 32 (41%) patients had continued antegrade flow at completion of the hypogastric artery embolization. When coils were placed at or in the bifurcation of the hypogastric artery, 12 of 22 (55%) experienced claudication. When coils were placed in the proximal hypogastric artery, one of 10 (10%) claudicated. CONCLUSION: It is probably not necessary to completely occlude antegrade flow in the hypogastric artery to prevent a distal endoleak. Buttock claudication is rare when coils are placed in the proximal hypogastric artery rather than at its bifurcation or in its branches. PMID- 10834488 TI - Stent-graft therapy for subclavian artery aneurysms and fistulas: single-center mid-term results. AB - PURPOSE: To evaluate the potential of covered stents to replace surgery in the treatment of subclavian artery aneurysms and traumatic injuries. MATERIALS AND METHODS: Nine patients (five men, four women; age range, 20-83 years; mean, 54 years) with subclavian artery aneurysms (n = 5) or fistulas (n = 4) were treated with stent-grafts. All devices used were custom-made, consisting of polytetrafluoroethylene (PTFE)-covered Palmaz (n = 5), Wallstent (n = 2), Z stents (n = 8), or a polyester-covered Z stent (n = 1). One patient was lost to follow-up after 2 months. All others were followed up with clinical evaluation, computed tomography (CT), and/or ultrasound. RESULTS: All devices were deployed successfully with exclusion of the aneurysms and fistulas. There were two procedure-related complications (22%), consisting of groin pseudoaneurysms requiring surgical repair 3 and 9 days after the procedure. One of those patients required additional oral antibiotic therapy for a postsurgical groin wound infection. One patient developed a stenosis at 12 months, which required angioplasty. The stent-graft thrombosed in one patient because of a kink 2 months after placement, which was successfully treated by thrombolysis and placement of a Wallstent. The primary and secondary patencies are 89% and 100%, respectively, after a mean follow-up of 29 months (2-66 mo). CONCLUSION: Mid-term results of stent-graft therapy of subclavian artery aneurysms and fistulas are encouraging, with low morbidity and excellent clinical outcome. PMID- 10834489 TI - Blue toe syndrome: treatment with intra-arterial stents and review of therapies. AB - PURPOSE: To determine if intra-arterial stent placement can adequately treat lesions producing microemboli to the lower extremities. MATERIALS AND METHODS: During a 6.5-year period, 15 patients presenting with blue toe syndrome had 16 presumed embolic lesions treated with intra-arterial stents. These patients were evaluated during routine clinical follow-up during a 6-month period. This evaluation included physical and noninvasive arterial examinations. When patients could not return for follow-up, hospital, clinical, vascular laboratory, and radiology records were reviewed to assemble the appropriate information. Outcomes included symptoms of recurrent emboli, amputation, and death. RESULTS: Treated embolic lesions included two aortic stenoses, three bilateral iliac artery stenoses, nine unilateral iliac artery stenoses (one patient received separate treatment of unilateral iliac lesions), and two superficial femoral artery stenoses. Patients were followed-up for a mean of 18 months. Eight of 15 patients (53%) were improved or stable without complications. There were eight negative outcomes experienced in seven patients. Three patients (20%) were deceased at follow-up. Four patients (27%) had undergone amputation; one transmetatarsal amputation and three below-the-knee amputations. Only one of these was related to progressive disease in the treated extremity (7%). One patient (7%) experienced recurrent embolic symptoms. Stents were patent in all patients. CONCLUSION: Patients with blue toe syndrome are at high risk of limb loss and mortality despite treatment. Intra-arterial stent placement provides an alternative to standard surgical treatment. Further studies are needed to define the optimum therapy. PMID- 10834490 TI - Endoluminal repair of peripheral arterial aneurysms: 4-year experience with the cragg endopro system I. AB - PURPOSE: To assess the efficiency and long-term patency of the Cragg EndoPro System I in patients with peripheral arterial aneurysms. MATERIALS AND METHODS: In 10 patients, 13 stent-grafts were used to treat 15 arterial aneurysms. Aneurysms were located in the common iliac (n = 4), superficial femoral (n = 4), popliteal (n = 3), and subclavian arteries (n = 2), and in a femoropopliteal bypass-graft (n = 2). Follow-up ranged between 2 and 46 months (mean, 36 months). Examination included clinical status, color-coded duplex sonography, computed tomography angiography, and intra-arterial digital subtraction angiography (DSA). RESULTS: Technical success was achieved in all patients. Primary patency was four of four in iliac vessels and three of nine in non-iliac vessels; secondary patency in noniliac vessels was four of nine. Repairs included one local lysis, four percutaneous transluminal angioplasties, one surgical thrombectomy, and one bypass surgery. Stent wire disintegration was detected in one of four iliac stent grafts and in seven of nine noniliac stent-grafts. In noniliac grafts, significant stenoses occurred in three of nine; occlusion occurred in five of nine. One complication at the iliac level was a vessel wall penetration at the proximal stent edge, with development of a new aneurysmal formation. No late endoleaks were found. CONCLUSION: Exclusion of peripheral arterial aneurysms with stent-grafts is feasible. Long-term results are excellent in iliac vessels. Mechanical weakness of the stent assembly and frequent re-stenoses or occlusions are significant drawbacks in noniliac vessels with low patency rates. PMID- 10834491 TI - Transcatheter embolization for acute lower gastrointestinal hemorrhage. AB - PURPOSE: The authors review their experience using transcatheter embolization in the treatment of acute lower gastrointestinal hemorrhage. MATERIALS AND METHODS: A retrospective review was conducted on 17 patients who underwent superselective transcatheter embolization for an acute lower gastrointestinal hemorrhage. All 17 patients were followed clinically 4 days to 60 months (mean, 18.5 months) after embolization for the presence of ischemia or for recurrent bleeding. In addition, 12 of 17 patients were examined 1 day to 12 months (mean, 38.8 months) after embolization by means of colonoscopy or by pathologic review. RESULTS: Transcatheter embolization achieved immediate hemostasis in 15 of 17 patients (88%) and was the definitive treatment in 76%. The other two patients underwent successful surgical resections after incomplete hemostasis of cecal lesions. Two patients of the 15, with initially successful embolizations, had recurrent hemorrhage within 30 days; both underwent further embolization with one failure. No intestinal infarction or stricture developed in the 15 patients who underwent successful embolization. CONCLUSIONS: The authors' experience supports the role of transcatheter embolization as a primary means of therapy for patients with an acute lower gastrointestinal hemorrhage. Their data further supports growing evidence that superselective embolization may be most efficacious in reducing complication rates. PMID- 10834492 TI - Successful embolization of collaterals from the ovarian artery during uterine artery embolization for fibroids: a case report. PMID- 10834494 TI - Temporary balloon occlusion of the internal iliac arteries for control of hemorrhage during cesarean hysterectomy in a patient with placenta previa and placenta increta. PMID- 10834495 TI - Angiographic diagnosis and transarterial embolization of iatrogenic ovarian artery injury. PMID- 10834493 TI - Adjunctive 3D US for achieving portal vein access during transjugular intrahepatic portosystemic shunt procedures. AB - PURPOSE: To evaluate the usefulness of information provided by three-dimensional ultrasound (3D US) and to determine whether 3D US decreased the number of passes required to obtain portal vein (PV) access during creation of transjugular intrahepatic portosystemic shunts (TIPS). MATERIALS AND METHODS: Intermittent 3D US volume acquisitions were obtained during creation of TIPS in 20 patients. Useful information provided by 3D US was tabulated. The number of passes required to achieve PV access was recorded and results were compared retrospectively to 25 patients who underwent TIPS without 3D US. RESULTS: 3D US documented that the operator's opinion of which hepatic vein had been selected was incorrect in nine patients (45%), detected unfavorable PV anatomy that required modification of equipment or technique in seven patients (35%), permitted estimation of the trajectory required to access the targeted PV in all patients (100%), assisted in selecting the optimal point along the hepatic vein for origination of the needle pass in 11 patients (55%), allowed avoidance of a large hepatocellular carcinoma in one patient (5%), and confirmed that access into the main PV was intrahepatic in four patients (20%). The mean number of needle passes decreased from 10.4 in the historic control group to 4.6 in the 3D US group (P = .0001). CONCLUSION: 3D US provided imaging information that detected technical errors and altered anatomy, and provided positional and directional information to significantly improve needle pass efficiency. PMID- 10834496 TI - Percutaneous cholecystoduodenostomy: a case report. PMID- 10834497 TI - Acute changes in aortic wall mechanical properties after stent placement in rabbits. AB - PURPOSE: To evaluate mechanical property changes after endovascular stent placement in small-diameter arteries. MATERIALS AND METHODS: Self-expanding stents (Wallstent) were placed in the infrarenal aorta of five New Zealand White rabbits via a surgical right femoral approach. Blood pressure changes (deltaP) were monitored in the aorta. Blood flow velocity was measured with a 20-MHz, pulsed Doppler probe (n = 4) to calculate the pulsatility index. Aortic diameter (dA) and diameter changes (delta(d)) were measured with a 20-MHz probe in echo tracking mode. Diameter compliance (Cd) and distensibility coefficient (DC) were calculated as Cd = 2(delta)d/(delta)P and DC = 2delta(d)/delta(P)/dA. RESULTS: Aortic diameter increased from 3.360 +/- 0.4033 mm to 4.020 +/- 0.3033 mm after stent placement at the stent level only. Compliance decreased from 77.644 +/- 24.306 mm kPa(-1) to 31.150 +/-8.245 x 10(-3) mm kPa(-1) at the stent level, and was then significantly lower than upstream (98.500 +/- 53.196 mm kPa(-1)) and down-stream (59.047 +/- 13.833 mm kPa(-1)). There was no significant change in pulsatility index. CONCLUSIONS: Endovascular stent placement produces a significant decrease in arterial wall compliance of the rabbit abdominal aorta. PMID- 10834498 TI - Suramin inhibits proliferation of human arterial smooth muscle cells in vitro: potential drug for prevention of restenosis by local drug delivery. AB - PURPOSE: Suramin is known to inhibit proliferation of various tumor cells. This study was performed to investigate the effect of suramin on proliferating human arterial smooth muscle cells (HASMC) and thus to examine its suitability for the prevention of restenosis. METHOD: Proliferation of HASMC was stimulated with human whole blood serum (HWBS), as well as with platelet-derived growth factor (PDGF) and basic fibroblast growth factor (bFGF). Proliferation was assessed by measuring DNA synthesis and cell viability was assessed by measuring mitochondrial activity. RESULTS: Suramin exerted a strong attenuating effect on the proliferation stimulators HWBS, PDGF, and bFGF. A suramin concentration of 0.5 mmol/L, which is approximately twice the dosage used for systemic application, was sufficient for complete neutralization of proliferation stimulation by 10% HWBS added to the cell culture medium. CONCLUSION: The authors' data demonstrate a strong proliferation inhibiting effect of suramin on HASMC in vitro. Because it is known to interact not only with one but with a multitude of relevant growth factors, these results make suramin a most interesting substance for local application for prevention of hyperplastic neointima formation. PMID- 10834499 TI - Physical properties of endovascular stents: an experimental comparison. AB - PURPOSE: Different endovascular stent types (AVE Bridge, AVE Bridge X, Memotherm, Palmaz Large, Palmaz Medium, Palmaz-Schatz Long-Medium, Perflex, S.MA.R.T., Symphony, and Wall-stent) of 4 cm length and 8 mm diameter were subjected to standardized physical tests. MATERIALS AND METHODS: The metal mass of each stent was assessed by weighing. The balloon-expandable stents were pneumatically tested for hoop strength. In self-expanding stents, radial resistive force and chronic outward force were determined with use of a loop test. Stent delivery system pushability was assessed in a crossover model. Stent radiopacity was analyzed quantitatively. RESULTS: The hoop strength of the balloon-expandable stents ranged from 15.8 N/cm (Perflex) to 28.9 N/cm (AVE Bridge X). The stent weight increased with greater hoop strength (Perflex, 0.046 g/cm vs. AVE Bridge X, 0.061 g(cm). The self-expanding stents had a radial resistive force between 0.39 N/cm (Wallstent) and 1.7 N/cm (Smart). The flexible balloon-expandable stents showed pushability values between 0.13/N (AVE Bridge) and 0.20/N (Perflex). The self expanding stents had flexibilities between 0.13/N (Memotherm) and 0.24/N (Symphony). Radiopacity assessed with use of a phantom simulating the iliac region ranged from 92 (Palmaz Large) to 115 (AVE Bridge) on a 256-point gray scale (0 = black, 256 = white). CONCLUSIONS: There is no stent with ideal physical properties. However, depending on the characteristics of the arterial lesion to be treated, the most appropriate stent can be chosen. PMID- 10834500 TI - SCVIR Annual Meeting Film Panel Session: diagnosis and discussion of case 5. Society of Cardiovascular and Interventional Radiology. PMID- 10834501 TI - SCVIR Annual Meeting Film Panel Session: diagnosis and discussion of case 8. Society of Cardiovascular and Interventional Radiology. PMID- 10834502 TI - An obscure formula in case of renal dysfunction. PMID- 10834503 TI - Recent advances in antigen-targeted therapy in non-Hodgkin's lymphoma. AB - Substantial advances in antigen-targeted lymphoma therapy have been achieved in recent years that make the use of monoclonal antibodies a highly attractive concept and promise further improvements in the clinical management of malignant lymphoma. The development of the chimeric anti-CD20 antibody IDEC-C2B8 (Rituximab) proved the concept of an effective therapy with a single unconjugated monoclonal antibody in lymphoma patients. Radioimmunoconjugates with myeloablative activity induced response rates of 80-100% in heavily pretreated patients. Progress in the genetic engineering of immunotoxins has improved the efficacy of these constructs. Ongoing prospective clinical trials will define the optimal use of these innovative therapeutic agents in patients with malignant lymphoma, and may establish therapeutic strategies with a high anti-lymphoma specificity and a low unspecific toxicity. PMID- 10834504 TI - Therapy of B-cell lymphomas with monoclonal antibodies and radioimmunoconjugates: the Seattle experience. AB - Monoclonal antibodies (mAbs) and radioimmunoconjugates targeting B-cell differentiation antigens have emerged as promising new treatments for patients with relapsed non-Hodgkin's lymphoma. This review focuses on our experience in Seattle over the past decade treating relapsed B-cell non-Hodgkin's lymphomas. In initial pilot studies, we administered escalating doses of the unmodified murine anti-CD20 mAb 1F5 to four patients with multiply relapsed lymphoma and-documented the dose-dependent penetration of antibody into the bone marrow and lymph nodes. The two patients who received the higher doses (1032 mg and 2380 mg) had remissions of brief duration, including one minor response lasting 2 weeks and one partial response lasting 6 weeks. Sequential phase-I and -II trials with myeloablative doses of iodine-131-mAbs have documented complete responses in 30 of 36 (83%) patients treated and, most importantly, many of these responses have been durable. A recent long-term follow-up study of the 29 patients treated with myeloablative doses of the I-131-labeled murine anti-B1 (anti-CD20) antibody has documented estimated overall and progression-free survival rates of 68% and 42%, respectively, with a median follow-up time of 42 months. To optimize the durability of responses, we are currently conducting a phase-I/II trial studying the toxicity and efficacy of I-131-anti-B1 antibody given in combination with high-dose etoposide and cyclophosphamide and autologous hematopoietic stem cell rescue. PMID- 10834505 TI - Intraoperative endogenous erythropoietin levels and changes in intravascular blood volume in healthy humans. AB - There is accumulating evidence of a relationship between changes in intravascular blood volume and endogenous erythropoietin (EPO) levels. In this study, eight healthy adult American Society of Anesthesiologists class-I patients due for prolonged elective surgery were randomised either to preoperative hypervolaemic haemodilution using hydroxyethyl starch, followed by intraoperative crystalloid infusion, or to standard intraoperative normovolaemic fluid balance management using crystalloids (control group). Electrolytes, creatinine, urea, osmolality, urine output and blood gases were monitored pre- and intraoperatively for 6 h, Comparable cardiopulmonary and renal homeostasis were maintained in both groups. We found that central venous pressure increased and EPO levels decreased, both significantly, in the hypervolaemic haemodilution group relative to controls. There were no significant intergroup changes in any other parameters. By controlling for other known determinants of EPO levels, our data indicate a relationship between EPO levels and changes in intravascular blood volume in humans, supporting the notion of EPO as a volume-regulated, and possibly volume regulating, hormone. PMID- 10834506 TI - Oxidative burst measurement in patients treated with cytostatics: influence of G CSF and role as a prognostic factor. AB - The ability to generate reactive oxygen species, the so-called oxidative burst, is essential for neutrophils to kill infectious micro-organisms. Flow cytometry was used to study oxidative burst prior to, during, and after cytostatic therapy. Seven patients were treated according to the DexaBEAM regimen with 12 cycles monitored. Four patients were treated according to the B-NHL regimen in which nine cycles were monitored. Ten healthy volunteers were chosen as a control group without any treatment. Neutrophils were collected from heparinized peripheral blood and were stimulated by phorbol-12-myristate-13-acetate (PMA), N-formyl methionyl-leucyl-phenylalanine (FMLP), and Escherichia coli. The oxidative burst was estimated by the amount of nonfluorescent dihydrorhodamine 123 converted to green fluorescent rhodamine 123. Measurements were done daily. The FMLP-induced burst was enhanced in patients before therapy as compared with the control group, whereas PMA-induced burst was decreased slightly. E. coli-, FMLP-, and PMA induced oxidative burst decreased in both groups during cytostatic therapy. E. coli-induced burst increased again within 2 days of G-CSF treatment in vivo. FMLP induced burst increased in the B-NHL group but decreased in the DexaBEAM group. In patients who have recovered from leukopenia the oxidative burst is still partly suppressed. PMA-induced oxidative burst measured at the start of therapy correlates with infectious complications. Thus, PMA-induced burst may be used as a simple method for evaluating the individual risk of infections during therapy. The results demonstrate the modulating effect of cytostatic drugs on the oxidative burst and may explain why some patients suffer from severe bacterial infections although the total number of granulocytes is normal. PMID- 10834507 TI - Aberrant expression of the major sialoglycoprotein (CD43) on the monocytes of patients with myelodysplastic syndromes. AB - CD43, a sialylated glycoprotein expressed on the surface of most hematopoietic cells, has been implicated in cell adhesion and signaling. The reduced expression of this antigen in patients with Wiscott-Aldrich syndrome, in which progressive immunodeficiency is a major problem, raised the question whether abnormal expression of this molecule could affect the susceptibility to infections in patients with myelodysplastic syndromes (MDS). We studied the expression of this antigen on the monocytes of ten patients with chronic myelomonocytic leukemia (CMML) and compared the results with 67 patients suffering from other MDS syndromes and with 18 healthy individuals. We chose this series as it plays an important role in MDS patients where in most cases the neutrophils are defective. We also examined the following antigens as indicative of activation and adhesion of the monocytes in these patients: CD11b, CD18, CD35, CD38, CD44, CD69. We found decreased expression of CD43 on the monocytes of the RA, RAS, RAEB, and RAEB-t patients compared with the CMML and controls. The other activation molecules studied were found to be upregulated, suggesting the existence of activated monocytes in these patients. The increased levels of soluble vascular cell adhesion molecule in these patients suggest vascular endothelial activation in the absence of infection. Further experiments are needed to investigate the significance of CD43 downregulation in these patients, its role in cell adherence and tissue migration, and the correlation of the phenomenon to the increased susceptibility to infections observed in these patients. PMID- 10834508 TI - Excellent disease eradication by myeloablative therapy and stem-cell transplantation in patients with acute myelogenous leukemia. AB - Between February 1982 and 1999, 118 consecutive patients (65 male, 53 female) with acute myelogenous leukemia (AML), with a median age of 35 years (range 17-56 years), received stem-cell grafts from a human leukocyte antigen-identical sibling (n = 71), one-antigen-mismatched family member (n=2), matched unrelated donor (n=15), one-antigen-mismatched unrelated donor (n = 4) or an autologous (n = 26) graft. At the time of transplant, 56 patients were in the first complete remission (CR), 27 in the second CR, 6 in untreated relapse, 17 in primary refractory, and 12 in refractory relapse. The French-American-British classification (FAB) subtypes were as follows: M1 (n=25), M2 (n=28), M3 (n=11), M4 (n =32), M5 (n=16), M6 (n = 6). For conditioning, most patients underwent total body irradiation-containing regimens. As of 28 February, 1999, probability of leukemia-free survival (LFS) is 58% for patients after related and 45% after unrelated stem-cell transplantation (SCT). The probability of LFS is 70% for patients given allogeneic transplants in the first CR compared with 33% for those beyond the first CR at SCT. In autologous stem-cell graft recipients, the probability of LFS is 37%. Transplant-related mortality was 28% after related, 20% after unrelated, and 4% after autologous SCT. Probability of relapse for patients given related-donor stem-cell grafts in the first CR and beyond the first CR is 30% and 67%, 55% after unrelated and 63% after autologous stem-cell grafting. Thus, myeloablative therapy followed by allogeneic stem-cell infusion has a high curative potential for patients with AML in remission and offers substantial benefits to patients in advanced disease. PMID- 10834509 TI - Regression of Epstein-Barr virus-associated lymphoproliferative disorders in patients with acquired immunodeficiency syndrome during therapy with foscarnet. AB - Epstein-Barr virus (EBV)-associated lymphoproliferative disorders frequently develop in patients with acquired immunodeficiency syndrome (AIDS). The lymphoproliferative disorders are usually treated with a combination of cytotoxic drugs, and virustatic medication is thought to be ineffective. However, we report about two AIDS patients with EBV-associated lymphoproliferative disorders not responding to a standard chemotherapy, who achieved complete tumour regression after virustatic therapy with foscarnet. PMID- 10834510 TI - An unusual case of leukemic non-Hodgkin's lymphoma with blastic transformation. AB - We report on a patient who was diagnosed as having B-cell chronic lymphocytic leukemia (CLL) with atypical morphology. Flow cytometry disclosed CD5, CD19, and CD23 positivity, an immunophenotype seen mostly in B-CLL. Histology of the spleen and bone marrow suggested a diagnosis of small lymphocytic lymphoma. Upon blastic transformation, only 3 years after the diagnosis had been made, unusual clinical and laboratory features emerged. Lymphoid blasts appeared in the peripheral blood, and the patient developed nodular infiltrates consisting of these blasts at recent venous puncture sites. The patient did not respond to chemotherapy and died. The lymphoid blasts in the peripheral blood were CD5-, CD19+, and CD23+ and harbored t(11;14) (q13;q32) and t(11;21)(p11;q21) translocations. To account for the possibility of two independent lymphoid malignancies, molecular genetic analyses were performed on samples from the spleen, bone marrow and a lymph node with the large-cell lymphoma, which showed identical clones in these tissues. This unusual case supports the idea that in leukemic non-Hodgkin's lymphoma, in addition to morphology, an accurate diagnostic workup requires immunophenotypic, cytogenetic, and molecular studies. PMID- 10834511 TI - T-cell lineage acute lymphoblastic leukemia with chromosome 5 abnormality in a patient with Crohn's disease and lipoid nephrosis. AB - We describe a 17-year-old patient with a documented history of Crohn's disease (CD) and of minimal-change nephrotic syndrome (MCNS) in whom a diagnosis of T cell acute lymphoblastic leukemia (ALL) was made. The diagnosis of ALL was established by morphological examination of bone-marrow aspirates and confirmed by means of immunophenotypic analysis showing the involvement of T-cell lineage leukemic cells. The lymphoid clone showed a karyotypic abnormality involving the long arm of chromosome 5 in a translocation (5;6). Few cases of CD complicated by ALL have been previously reported. The present one is the first case combining CD and ALL in a patient with a past history of MCNS. This raises the possibility of a relationship among those diseases. The possible mechanisms for such a relationship are discussed here. PMID- 10834512 TI - Identification of novel mutations in Arabs with cystic fibrosis and their impact on the cystic fibrosis transmembrane regulator mutation detection rate in Arab populations. AB - The cystic fibrosis transmembrane regulator (CFTR) gene in Arab patients with cystic fibrosis (CF) (sweat chloride > 60 mmol/l) from 61 unrelated families was screened for mutations in exons 3, 4, 5, 7, 10, 11, 16 and 19 and for mutations W1282X, N1303K and 3,849 + 10kbC --> T. Eight novel mutations were identified. These are: in exon 4: a) 425del42 (an in-frame 42 bp deletion that removes 14 amino acids and causes Gln98 --> His at the point of deletion), b) 475G --> T (Glu115 --> Stop) and c) 548A --> T (His139 --> Leu); in intron 5,711 + 1G --> A (splice site mutation); in exon 10, 1548delG (deletion of a "G" nucleotide causing a frameshift mutation that alters the amino acid sequence at residue 473 and results in translation termination at residue 526); in exon 11, a) 1729T --> C (Ph533E --> Leu) and b) 1,811 + 2 (splice site mutation) and finally in exon 19,3361A --> T (Lys1177 --> Stop). All mutations were detected by heteroduplex analysis and identified by sequencing. Of more than 850 known CFTR mutations, only 9 were encountered. The comparative frequencies of the most common mutations are: 1548delG> 1123V = deltaF508 = 3,120 + 1G --> A > H139L. Screening for these five mutations identifies 60% of the CF alleles in Arab populations. The novel mutation 1548delG is the most frequent (17%) among Arabs. CONCLUSION: Novel Arab specific mutations were identified in the CFTR gene underlying cystic fibrosis. As a result of this study, the CFTR mutation detection rate among Arabs with cystic fibrosis is now comparable to that of other populations. PMID- 10834513 TI - Auto-antibodies to the asialoglycoprotein receptor in sera of children with auto immune hepatitis. AB - Most studies on the prevalence of asialoglycoprotein antibodies have involved adults. In this study the prevalence of antibodies to the human asialoglycoprotein receptor (ASGP-R) was determined in 63 children with auto immune hepatitis. It was shown that 75% of those with auto-immune hepatitis type 1 and 40% of those with auto-immune hepatitis type 2 were positive and the presence of anti-ASGP-R auto-antibodies, at a mean titre of 1:1,600 and 1:1,000 respectively. No statistical significance was found between anti-ASGP-R positive and negative patients with respect to the median age at onset, proportion of females and ALT levels. However, the titre of anti-ASGP-R antibodies correlated significantly with hypergammaglobulinaemia. CONCLUSION: Anti-asialoglycoprotein receptor antibodies are frequently present in the sera of children with auto immune hepatitis, particularly type 1, The presence of these antibodies is an indirect marker of inflammatory activity. PMID- 10834514 TI - How many forms of glycogen storage disease type I? AB - Glucose-6-phosphatase is a multicomponent enzymatic system of the endoplasmic reticulum, which catalyses the terminal steps of gluconeogenesis and glycogenolysis by converting glucose-6-phosphate to glucose and inorganic phosphate. Glycogen storage diseases type I (GSD I) are a group of metabolic disorders arising from a defect in a component of this enzymatic system, i.e. the glucose-6-phosphate hydrolase (GSD Ia), the glucose-6-phosphate translocase (GSD Ib) and possibly also the translocases for inorganic phosphate (GSD Ic) or glucose (GSD Id). The genes encoding the glucose-6-phosphate hydrolase and the glucose-6-phosphate translocase have both been cloned and assigned to human chromosomes 17q21 and 11q23, respectively. Investigation of patients with GSD I shows that those with GSD Ia are mutated in the glucose-6-phosphate hydrolase gene, whereas those diagnosed as GSD Ib, GSD Ic or GSD Id are mutated in the glucose-6-phosphate translocase gene, and are therefore GSD Ib patients, in agreement with the fact that they all have neutropenia or neutrophil dysfunction. This suggests that the biochemical assays used to differentiate GSD Ic and GSD Id from GSD Ib are not reliable. CONCLUSION: In practice therefore appears to be only two types of GSD I (Ia and Ib), which can be differentiated by (1) measurement of glucose-6-phosphatase activity in fresh and detergent-treated homogenates and (2) by mutation search in the genes encoding the glucose-6 phosphate hydrolase and the glucose-6-phosphate translocase. PMID- 10834515 TI - Influenza A virus encephalopathy with symmetrical thalamic lesions. AB - During an epidemic of influenza A infection in Japan, a 7-year-old boy was admitted to our hospital because of high fever, convulsions, coma, and liver dysfunction on the 2nd day of a cold-like illness. His serum CPK was markedly elevated, but there was no hyperammonaemia or hypoglycaemia. His CSF showed an increased protein level, but the cell count and glucose level were normal. CT and MRI of the brain showed symmetrical thalamic lesions, and he was diagnosed with acute necrotizing encephalopathy in childhood. He had a significant increased in antibodies to influenza A H1N1 in serum and CSF, but the CSF was negative for influenza virus using virus isolation and a polymerase chain reaction assay. CONCLUSION: Antibody production without detectable levels of influenza virus in cerebrospinal fluid suggests that virus infection occurred, but the virus did not replicate in sufficient numbers in his central nervous system. The thalamic lesion, the hallmark of acute necrotizing encephalopathy in childhood, may be initiated by a local virus infection and develop with subsequent local changes such as breakdown of the blood-brain barrier and the extravasation of blood. PMID- 10834516 TI - Glycogen storage disease type Ia: recent experience with mutation analysis, a summary of mutations reported in the literature and a newly developed diagnostic flow chart. AB - We studied the glucose-6-phosphatase (G6Pase) gene of 30 unrelated glycogen storage disease type Ia (GSD Ia) patients using single strand conformational polymorphism (SSCP) prior to automated sequencing of exons revealing an aberrant SSCP pattern. In all patients we could identify mutations on both alleles of the G6Pase gene, indicating that this method is a reliable procedure. A total of 14 different mutations were identified. R83C (16/60), 158delC (12/60), Q347X (7/60), R170X (6/60) and deltaF327 (4/60) were found most frequently. Nine other mutations accounted for the other 15 mutant alleles. Two DNA-based prenatal diagnoses were performed successfully. At present, 56 mutations in the G6Pase gene have been reported in 300 unrelated GSD Ia patients and an overview of these mutations is presented. Evidence for a clear genotype-phenotype correlation could be established neither from our data nor from those in the literature. With increased knowledge about the genetic basis of GSD Ia and GSD Ib and the high detection rate of mutations, it is our opinion that the diagnoses GSD Ia and GSD Ib can usually be based on clinical and biochemical abnormalities combined with mutation analysis instead of enzyme assays in liver tissue obtained by biopsy. A newly developed flowchart for the diagnosis of GSD I is presented. CONCLUSION: Increased knowledge of the genetic basis of glycogen storage disease type I provides a DNA-based diagnosis, prenatal DNA-based diagnosis in chorionic villus samples and carrier detection. PMID- 10834517 TI - Safety and efficacy of interventional occlusion of patent ductus arteriosus with detachable coils: a multicentre experience. AB - In order to define the safety and efficacy of Cook detachable coils for interventional closure of patent ductus arteriosus, we performed a retrospective analysis of all patients who underwent cardiac catheterization in seven centres for intended interventional occlusion of patent arterial duct. From January 1995 until March 1998, cardiac catheterization for intended interventional occlusion of patent arterial duct was performed in 317 consecutive children. Successful placement of at least one coil was achieved in 282 children (89%). The mean diameter of the ductus in children treated with Cook detachable coils was 1.65 mm, mean fluoroscopy time was 10.7 min. Occlusion rates were 62% 10 min after the procedure, 82% at the time of discharge, 91% at 4 months and 95% at late follow up (2 years). In children with a ductus diameter of < or = 2.5 mm the rate of successful coil deployment was 94% with a 98% occlusion rate at late follow-up. Complications occurred in 11 procedures (3.5%) including haemolysis (3 patients), embolization of a coil to the pulmonary artery (7 patients, 2.3%) and inability to release a coil (1 patient). CONCLUSION: In our opinion, Cook detachable coils are safe and effective especially in the treatment of persistent ductus arteriosus with a diameter < or = 2.5 mm. Due to the low costs these coils appear to be superior to other devices in this subgroup of patients. PMID- 10834518 TI - Predisposing factors in childhood masturbation in Turkey. AB - The aim of this prospective, referral-based study was to assess demographical and developmental features associated with childhood masturbation in Turkey. A total of 61 children with childhood masturbation who were referred for the first time to the Department of Child Psychiatry were examined from demographical, psychosocial and medical aspects and compared with two control groups consisting of 61 age and gender matched children who were brought to the paediatric outpatient clinics and 43 children and adolescents who were the biological siblings of the study group. In children with masturbation, sleep difficulties were more frequent (P < 0.001) and breast-feeding was shorter than in controls (P < 0.05). The onset of masturbation was often associated with a genito-urinary disorder or a stressful life event like weaning, the birth of a sibling, or separation from the parents. CONCLUSION: This is the first controlled study investigating the clinical and the developmental features of childhood masturbation. These findings may help identify children who could be at risk for this condition. PMID- 10834519 TI - Differences in management of bronchiolitis between hospitals in The Netherlands. Dutch Paediatric Respiratory Society. AB - To study the variation in management of acute bronchiolitis in the Netherlands, a questionnaire on the use of diagnostic and therapeutic procedures and prescription of drugs after discharge was mailed to all 110 hospital-based paediatric practices in the Netherlands. A 100% response rate was achieved. There was a great deal of variation in management of bronchiolitis between respondents. The most commonly applied diagnostic procedures were immunofluorescence staining of nasopharyngeal secretions for respiratory syncytial virus (100% of respondents), blood gas analysis (93%), leucocyte count/differentiation and serum C-reactive protein levels (92%), and chest X-rays (83%). Most respondents used supplemental oxygen (100%) and tube feeding (96%) when needed, and gave nebulized bronchodilators, either as a trial (59%) or in a fixed schedule (33%). Antibiotics for suspected bacterial co-infection were used by 69% of respondents. Corticosteroids were used for severe cases by 35% of respondents; ribavirin was only used in 11% of hospitals for treatment of children from high-risk groups. When children had responded favourably to bronchodilators during admission, these drugs were continued after discharge by 69% of respondents; more than half of these also prescribed inhaled corticosteroids to these children. CONCLUSION: Considerable variation in management of bronchiolitis exists between hospitals in the Netherlands. Several diagnostic and therapeutic approaches are used which are not evidence based, probably reflecting the lack of therapeutic options with proven clinical efficacy for this condition. PMID- 10834520 TI - Growth, puberty and hypothalamic-pituitary function in children with suprasellar arachnoid cyst. AB - A suprasellar arachnoid cyst may cause disorders of growth, puberty and hypothalamic-pituitary function, due to the proximity of the cyst to the hypothalamic-pituitary area. A total of 30 patients (17 boys) with cyst diagnosed at 4.3 +/- 1 years were routinely evaluated at 5.4 +/- 1 years; 24 of them had one or multiple cyst derivations. Some 23 cases had an abnormal height, weight or puberty: short (< -2SD, 5 cases) or tall ( > 2SD, 10 cases) stature, overweight (body mass index, BMI, > 2SD, 6 cases), central precocious puberty (10 cases) and/or no progression of pubertal development (3 cases). The growth hormone (GH) peaks after pharmacological stimulation test were low (< 10 MICROg/L) in 16 patients, confirmed by a second evaluation in 8/11 of them. The plasma free thyroxine was low in five patients, prolactin was high in two and the cortisol and concomitant plasma and urinary osmolalities were normal. BMI was correlated negatively with the GH peaks (r = -0.37, P < 0.01) and positively with the plasma leptin concentrations (r = 0.55, P < 0.01). The plasma fasting insulin concentrations were also correlated negatively with the GH peaks (r = -0.55, P < 0.02) and positively with the plasma insulin-like growth factor I concentrations (r = 0.64, P < 0.002). The adult height (12 cases) was at 4SD in 1 and < -2SD in 4 patients, two of whom had precocious puberty untreated with gonadotropin releasing hormone (GnRH) analogue, and two had untreated GH deficiency. The adult height of those treated was normal. One girl had primary amenorrhoea and two boys had low plasma testosterone, despite a normal gonadotropin response to a GnRH test. CONCLUSION: Suprasellar arachnoid cysts may cause deficiencies of growth hormone and thyrotropin, stimulation of the hypothalamic-pituitary-gonadal axis, tall stature and/or overweight. These last two disorders may be due to hyperinsulinism, itself due to suprasellar arachnoid cyst. PMID- 10834521 TI - Polymerase chain reaction detection of Bartonella henselae bacteraemia in an immunocompetent child with cat-scratch disease. AB - A case of Bartonella henselae bacteraemia is reported in an immunocompetent 8 year-old boy with cat-scratch disease. Serology to B. henselae, diagnosed by polymerase chain reaction, was positive. DNA was extracted from peripheral whole blood and amplified with specific primers targeting the htrA gene of B. henselae. A non-isotopic hybridization assay with a species-specific oligonucleotide probe was used to detect the amplified product. CONCLUSION: The polymerase chain reaction can be used for the rapid laboratory diagnosis of bacteraemia in cat scratch disease. PMID- 10834522 TI - Cyclic vomiting syndrome and food allergy/intolerance in seven children: a possible association. AB - Cyclic vomiting syndrome (CVS) is characterized by repeated unpredictable, explosive and unexplained bouts of vomiting. The episodes have a rapid onset, persist over a number of hours or days, and are separated by symptom-free intervals. Despite the recent interest in this disorder, its aetiology, pathogenesis and even its target organ remain unknown. The purpose of this study is to investigate the role played by food allergy in CVS. The report concerns eight children (five male, three female), mean age 8 years (3-13 years), suffering from CVS for 2 years at least. The diagnosis of CVS was based on characteristic history, normal physical examination and negative laboratory, radiographic, neurological and endoscopic studies. Despite the absence of clinical signs typical of food allergy, skin prick tests were positive in six of the eight patients (75%). Specific IgE were present in 4/8 (50%) of the patients. Skin tests and specific IgE were positive for cow's milk proteins, egg white and soya. IgE levels were higher than the mean + 2SD in 5/8 (63%) of the patients. A double blind placebo controlled food challenge (DBPCFC) was carried out on seven of the eight patients who displayed clinical improvement after an elimination diet for cow's milk (and other foodstuffs indicated by positive skin tests). The DBPCFC was positive in all seven children. Clinical follow-up revealed a state of well-being over the 6 months of observation. CONCLUSION: It appears reasonable to suggest that food allergy plays a role in cyclic vomiting syndrome. PMID- 10834523 TI - Prophylactic ibuprofen therapy of patent ductus arteriosus in preterm infants. AB - This study was aimed at evaluating the efficacy of ibuprofen in the prophylaxis of patent ductus arteriosus (PDA) in very preterm neonates and at detecting eventual side-effects. A total of 46 preterm neonates with gestational age under 31 weeks were randomly assigned at 2 h of life: 23 to the prophylaxis group and 23 to the control group. The prophylaxis group received intravenous treatment with ibuprofen lysine (10 mg/kg), followed by 5 mg/kg after 24 h and 48 h. No placebo was given to the control group. No PDA was demonstrated at 72 h of life in 20 of the 23 babies in the ibuprofen group (87%) nor in 7 of the 23 control neonates (30.4%). All neonates with PDA received treatment with indomethacin. One neonate in the prophylaxis group and three in the control group underwent surgical ligation. Prophylaxis with ibuprofen was not associated with any significant side-effect except for food intolerance. CONCLUSION: Ibuprofen prophylaxis seems to be efficient in closing patent ductus arteriosus and in reducing indomethacin treatment. No significant early side-effects were found due to ibuprofen. PMID- 10834524 TI - Final height, gonadal function and bone mineral density of adolescent males with central precocious puberty after therapy with gonadotropin-releasing hormone analogues. AB - Few data are available on the outcome of boys with central precocious puberty (CPP) treated with gonadotropin-releasing hormone (GnRH) analogues. We report on final height, endocrine and exocrine testicular function, and bone mineral density (BMD) in nine males (age 16.7 +/- 1.5 years) treated with GnRH analogues from the age 6.0 +/- 1.8 years for a mean period of 5.6 +/- 2.4 years. The following parameters were evaluated: final height, serum gonadotropin and gonadal steroid levels, spermarche, semen analysis, area and volumetric BMD. Final height (-0.4 +/- 1.1 SDS) was significantly higher than pre-treatment predicted adult height (-2.0 +/- 1.2 SDS) and not significantly different than midparental height (-0.1 +/- 0.8 SDS). Pubertal response of gonadotropins to GnRH test occurred within 1.5 years (mean 0.7 +/- 0.4 years) and spermarche (n = 7) from 0.7 to 3 years (1.8 +/- 0.9 years) after the discontinuation of GnRH analogue therapy. No alteration in semen analysis was found (n = 6, sperm count, 10(6)/ml: 52.0 +/- 18.7; normal motility (%): 49.5 +/- 18.7; atypical morphology (%): 44.5 +/- 11.4). Area and volumetric BMD were not reduced (0.2 +/- 1.0 SDS and -0.1 +/- 0.9 SDS, respectively). CONCLUSION: Long-term treatment with gonadotropin-releasing hormone analogues improves final height in boys with central precocious puberty. Post-therapy data demonstrating normal endocrine and exocrine testicular function support the safety of gonadotropin-releasing hormone analogues on reproductive function. Long-term pharmacological suppression of testicular function in childhood does not impair bone mineral density in late adolescence. PMID- 10834525 TI - Severe hypercalcaemia and respiratory insufficiency associated with infantile hypophosphatasia caused by two novel mutations of the tissue-nonspecific alkaline phosphatase gene. AB - We report the case of a male patient with infantile hypophosphatasia associated with severe hypercalcaemia and mild respiratory insufficiency. At the age of 2 months, severe hypercalcaemia, low levels of serum alkaline phosphatase activity, and elevated urinary excretion of calcium and phosphoethanolamine were noted. Radiological findings showed generalized osteopenia and disturbed and irregular ossification of the metaphyses. Their involvement had spontaneously improved at the age of 6 months. A genetic study revealed that the tissue-nonspecific alkaline phosphatase gene of the patient had two novel mutations, K207E and G409C, derived from the mother and father, respectively. A reconsitution experiment revealed that both mutant gene products had low but significant enzymatic activity. CONCLUSION: The detection of tissue-nonspecific alkaline phosphatase gene mutations and expression studies to determine the enzymatic activity of mutant gene products was useful for assessing the clinical course of this patient with hypophosphatasia. PMID- 10834526 TI - The effect of dexamethasone on growth, mineral balance and bone mineralisation in preterm infants with chronic lung disease. AB - The aim of this prospective longitudinal study was to observe the effects of treatment with dexamethasone on somatic growth, mineral balance and bone mineralisation in very low birth weight (VLBW) preterm infants with chronic lung disease (CLD). Dexamethasone was started at a dose of 500 microg/kg body weight per day for 3 days followed by gradually decreasing doses for a total period of 3 weeks' treatment if the infant was still mechanically ventilated in the 3rd postnatal week and had signs of CLD on the chest radiograph. Eleven infants with CLD treated with dexamethasone were studied. Administration of dexamethasone treatment was associated with significantly (P < 0.05) lower weight velocity, head circumference velocity, lower leg length (measured by the neonatal knemometer) velocity compared to pre-treatment changes. The start of dexamethasone treatment was also associated with a fall in calcium absorption (61% to 41.7%, P < 0.05) calcium retention (60.8% to 40.6%, P < 0.05) and phosphate retention (65% to 39.6%, P < 0.05); phosphate absorption was not significantly affected (88.8% to 92%, P > 0.05). Somatic growth and mineral balance improved during the immediate post-treatment period. Acute disturbances of bone mineral content (measured by dual energy radiographic densitometry), plasma calcium and phosphate were also seen but not reaching statistical significance at the P < 0.05 level. CONCLUSION: The start of steroids is associated with a rapid and significant fall in growth velocity, calcium absorption and calcium and phosphate retention in infants with chronic lung disease with recovery occurring after completion of steroid treatment. PMID- 10834527 TI - Ulcero-necrotic cutaneous lesions in an infant. PMID- 10834528 TI - Association between hyperproinsulinemia and anti-GAD65 antibodies in a patient with new onset type I diabetes. PMID- 10834529 TI - Anthracycline induced cardiomyopathy: successful treatment with angiotensin converting enzyme inhibitors. PMID- 10834530 TI - Prostate-specific antigen as a marker of prostate disease. AB - Serum prostate markers, in particular prostate-specific antigen (PSA), have truly revolutionised all aspects of the management of men with prostatic carcinoma (PCa), the most important application being related to its early detection and screening. Several studies have shown the clinical utility of PSA levels for staging patients with PCa, especially when associated with other parameters, such as tumour grade, digital rectal examination and transrectal ultrasound findings, to establish the likelihood of disease extension outside the gland and of positive lymph nodes. Also, serum PSA levels are useful in monitoring patients either after the initial diagnosis of PCa or following therapy. PMID- 10834531 TI - Sclerosing, pseudovascular rhabdomyosarcoma in adults. Clinicopathological and immunohistochemical analysis of three cases. AB - Rhabdomyosarcoma in adults represents a rare soft tissue neoplasm which is seen most frequently in its pleomorphic subtype in this age group. Very rarely, clear cell and spindle-cell variants have been reported. In this study we describe three cases of rhabdomyosarcoma in adult patients, characterised by prominent hyaline sclerosis and a pseudovascular growth pattern. All cases were identified in the consultation files of one of the authors and routinely processed. Immunohistochemical studies were performed on paraffin sections with the alkaline phosphatase-antialkaline phosphatase method. The patients, two women and one man, were 40, 41, and 56 years old. One developed a deep-seated soft tissue mass in the left lower leg, and one, a tumour of the left upper jaw. In one patient a bone tumour in the proximal body of the sacrum without extension into soft tissues was seen. The patients were treated by wide excision, piecemeal excision and incomplete excision in one case each; additional radiotherapy was performed in all three cases, and chemotherapy in two patients. In one patient multiple pulmonary metastases were noted, which showed progression despite systemic chemotherapy. Histologically, the neoplasms were composed of round/polygonal and spindle-shaped tumour cells including typical rhabdomyoblasts. In all cases a pseudovascular pattern and prominent hyaline sclerosis of the intercellular matrix was seen. Immunohistochemically, tumour cells stained positively for desmin and muscle actin (HHF35) and also for markers of striated muscle differentiation (myogenin, MyoD1, fast myosin). In this paper an unusual morphological variant of rhabdomyosarcoma arising in adult patients is described, which should be added to the morphological spectrum of these neoplasms. PMID- 10834532 TI - Aberrations of chromosomes 5 and 8 as recurrent cytogenetic events in anaplastic carcinoma of the thyroid as detected by fluorescence in situ hybridisation and comparative genomic hybridisation. AB - Comparative genomic hybridisation (CGH) is a technique which identifies gains and losses of DNA sequence copy number in tumours. We used CGH to search for genetic changes in one of the most aggressive malignancies--anaplastic thyroid carcinoma (ATC). For this purpose, we analysed tumour specimens of nine ATCs and DNA of two ATC cell lines. CGH detected aberrations in 10 of 11 samples, with a mean number of gains or losses per carcinoma of 4.8 (range 0-13). Total or partial changes of chromosome 8 (n=6), including gains or losses of 8p (n=6) or 8q (n=5) were those detected most frequently. Chromosome 5p was amplified in five cases. Gains in two of three samples were found for 3q, 7p, 11q and 20q. Gains in a fewer number were seen for 1p (1 case), 1q (1), 7q (2), 9q (2), 11p (2), 12q (1), 14 (1), 15 (1), 17q (2), 18p (2), 18q (1), 20p (1), 21 (2), Xp (2) and Xq (2). Losses were less frequent than gains and observed for 1p (2 cases), 1q (1), 2p (1), 2q (2), 3p (2), 3q (1), 4q (2), 6q (1), 9p (2), 9q (1), 18p (1), 18q (1) and Y (2). Examples of analysis of tumour sections and cell lines performed by fluorescence in situ hybridisation (FISH) confirmed the gains and losses found by CGH and detected additional signals for 8q21 in tumour cells in a sample with no gains or losses normally in CGH. The results suggest that aberrations of 5p, 8p and 8q, which are rarely found in differentiated thyroid carcinoma, may play an important role in the development of ATC. Therefore, these chromosomes could harbour gene loci potentially involved in the aggressiveness of neoplastic tumours, as shown in tumours such as in this study for ATC. PMID- 10834533 TI - Immunoexpression of transforming growth factor beta in desmoplastic ameloblastoma. AB - Desmoplastic ameloblastoma (DA) is an unusual subtype of ameloblastoma histologically characterized by the pronounced collagenized stroma. In the present study, the immunolocalization of transforming growth factor beta (TGF beta), one of the most potent local factors for modulating extracellular matrix formation, was observed in DA in order to study its participation in the stromal desmoplasia. Seven cases of DA, including a "hybrid" lesion, were studied together with ten cases of ordinary follicular and plexiform ameloblastomas as the control. In contrast to ordinary ameloblastomas, marked immunoexpression was observed in all DAs but one. In the "hybrid" lesion, TGF-beta was not expressed in the area of follicular ameloblastoma but in that of DA. These results show that TGF-beta produced by tumor cells of DA plays a part in the desmoplastic matrix formation. PMID- 10834534 TI - Immunohistochemical demonstration of an enamel sheath protein, sheathlin, in odontogenic tumors. AB - Enamel proteins can be useful markers for assessment of the functional differentiation of neoplastic epithelium and the nature of extracellular matrices in odontogenic tumors. In the present study, we examined immunohistochemical localization of sheathlin, a recently cloned enamel sheath protein, in various odontogenic tumors to evaluate functional differentiation of tumor cells and the nature of hyalinous or calcified matrices in odontogenic neoplasms. Distinct immunolocalization of sheathlin was observed in the immature enamel of the tooth germ at the late bell stage. Secretory ameloblasts facing the enamel matrix also showed positive staining in their cytoplasm. Definite localization of sheathlin was demonstrated in the enamel matrix in odontogenic tumors with inductive dental hard tissue formation such as ameloblastic fibroodontomas and odontomas. Immunoexpression of sheathlin was, furthermore, demonstrated in eosinophilic droplets in solid nests of adenomatoid odontogenic tumor (AOT) and ghost cells in the epithelial lining of calcifying odontogenic cyst (COC). In AOT, cells facing the eosinophilic droplets also expressed the protein in their cytoplasm. There was neither intracellular staining for sheathlin in the tumor cells nor extracellular staining in the matrix of ameloblastomas and calcifying epithelial odontogenic tumors. Dentin, dysplastic dentin-like hyaline material and cementum in the tumors examined were negative for sheathlin. These results show that immunodetection of sheathlin is a useful marker for functional differentiation of secretory ameloblasts and enamel matrix, which is often hard to differentiate from other hard tissues in odontogenic tumors. Our findings from the view point of sheathlin expression support that the tumor cells of ameloblastomas do not attain full differentiation into functional ameloblasts. It is very interesting that epithelial cells in odontogenic tumors can differentiate into functional ameloblasts without induction by odontogenic mesenchyme, as shown by immunoexpression of sheathlin in eosinophilic droplets within solid epithelial sheets in AOT and ghost cells in the epithelial lining of COC where inductive participation of mesenchymal cells was most unlikely. PMID- 10834535 TI - Differential expression of CD44s and CD44v3-10 in adenocarcinoma cells and reactive mesothelial cells in effusions. AB - The detection of malignant cells in serous effusions obtained from patients diagnosed with cancer marks the presence of metastatic disease and is associated with a poor outcome. The purpose of this study was to evaluate the role of CD44s and CD44v isoforms in the distinction between mesothelial cells and malignant epithelial cells in effusions. Fifty-nine fresh pleural and peritoneal effusions were studied. These consisted of 41 specimens from patients with known gynecological neoplasms, 9 from patients diagnosed with breast adenocarcinoma, and 9 effusions from patients with various nongynecological malignancies or tumors of unknown origin. Forty-three effusions contained malignant/atypical epithelial cells, and 16 effusions were diagnosed as reactive. Three effusions contained exclusively malignant cells. Specimens were stained with anti-CD44s, v3, v5, v6, v7 and v3-10. The presence of staining in cancer cells, benign mesothelial cells and lymphocytes was evaluated. CD44s immunoreactivity was seen in 10 of 43 (23%) cases in malignant/atypical epithelial cells and in 53 of 56 (94%) cases in benign cells. In contrast, CD44v3-10 was seen in 23 of 43 (55%) cases in malignant/atypical epithelial cells and in 3 of 56 (6%) cases in benign cells. We advocate the use of CD44s and CD44v3-10 immunostaining in diagnostic evaluation of difficult serous effusions. PMID- 10834536 TI - Plasmacytoma of the tonsil with AL amyloidosis: evidence of post-fibrillogenic proteolysis of the fibril protein. AB - We report of a 58-year-old Caucasian man who was referred to the University Hospital with a greatly enlarged left tonsil which showed calcifications on computed-tomography scans. Histopathology revealed a plasmacytoma with secondary AL amyloidosis, ossifications, and multinucleated foreign-body-type giant cells. N-terminal sequencing of amyloid-fibril proteins purified from the formalin-fixed tissue showed the presence of two proteins of different size; these were of lambda-light-chain origin (subgroup V), measured approximately 15.2 kDa and 10.5 kDa, and had identical N-terminal ends (YVLTQPP). When the amyloid deposits were immunolabeled with a polyclonal antibody directed against lambda light chain, they showed two staining patterns: some deposits showed intense immunolabeling while others were not immunoreactive. Immunostaining of amyloid was completely absent after protease pre-treatment. Immunoelectron microscopy with gold-labeled secondary antibodies showed staining that was spatially related to amyloid fibrils and suggested that the antibody probably detected the fibril protein. Therefore, our hypothesis in this case is that the different immunostaining patterns are due to a post-fibrillogenic proteolysis of the fibril protein at the C-terminal end of the light chain, as indicated by the presence of two differently sized lambda-light-chain fragments with identical N-terminal ends. PMID- 10834537 TI - Familial amyloidotic polyneuropathy (ATTR Ser50Ile): the first autopsy case report. AB - We report an autopsy case of a pedigree of familial amyloidotic polyneuropathy (FAP) with a mutation of isoleucine-50 transthyretin (ATTR Ser50Ile). A 47-year old man started developing severe diarrhea and weight loss at age 41 years, followed by urinary incontinence, autonomic-nervous-system abnormalities and serious heart failure; the diagnosis of FAP (ATTR Ser50Ile) was made on the basis of genetic, histochemical and immunohistochemical analysis. Six years after the initial symptoms, he died of septic shock. Autopsy revealed suppurative peritonitis, perforation of the sigmoid colon and marked systemic amyloid deposition. The total amount of amyloid deposited in the heart was greatly increased and was much lower in the thyroid gland and kidneys compared with amyloid deposits in ordinary FAP (ATTR Val30Met). Amyloid deposition in peripheral vessel walls was prominent, particularly in lymphatics and veins. His elder sister, 54 years old, started to develop orthostatic hypotension at age 49 years, followed by dysesthesia, diarrhea and severe congestive heart failure. Endomyocardial biopsy revealed severe TTR-amyloid deposition; ultrastructural examination demonstrated that amyloid fibrils were deposited disproportionately and extended radially around microvessels. PMID- 10834538 TI - Expression of vascular endothelial growth factor in renal cell carcinomas. AB - Vascular endothelial growth factor (VEGF) is an angiogenic factor that may be involved in tumor growth and metastasis. Only a few data concerning the role of VEGF in renal cell carcinomas (RCCs) are available, and no studies have yet evaluated its prognostic value. The aim of the present study was to assess VEGF expression in a large series of renal tumors with a long follow-up, correlated with the usual histoprognostic factors and survival. VEGF immunostaining was performed on formalin-fixed, paraffin-embedded archival tissue from 74 renal carcinomas (62 conventional renal cell and 12 papillary carcinomas). Positivity of immunostaining was semi-quantitatively scored by two pathologists. Angiogenesis was evaluated by immunostaining with anti-CD34 antibodies on serial sections. Cytoplasmic VEGF expression was detected in tumor cells in 35% (26/74) of RCCs, including 18 out of the 62 (29%) conventional RCCs and 8 out of the 12 (67%) papillary carcinomas (P=0.02). In the group of conventional RCCs, VEGF expression was positively correlated with both nuclear grade (P=0.05) and size of the tumor (P=0.05). Furthermore, a significant correlation was observed between VEGF expression and microvascular count (P=0.04). Finally, cumulative survival rate was significantly lower in the group of patients with conventional RCCs expressing VEGF (log rank test, P=0.01). In the Cox model, VEGF expression was a significant independent predictor of outcome, as well as stage and nuclear grade. This study suggests that VEGF is involved in angiogenesis in conventional RCCs and appears to be a potential prognostic factor in these tumors. PMID- 10834539 TI - Primary gastric apoptosis-rich T-cell lymphoma co-expressing CD4, CD8, and cytotoxic molecules. AB - In contrast to primary gastric lymphomas of B-cell type, little is known about primary gastric T-cell lymphomas. We describe three cases with remarkably similar features: diffuse growth, epitheliotropism, medium too large cell size, high apoptotic rates, and a CD3+, CD4+, CD8+, CD45RO+ immunophenotype. Clonal TCRgamma gene rearrangement was shown in two cases. Epstein-Barr virus infection was excluded in two cases. Taking advantage of fresh-frozen material, we analyzed two cases further, revealing CD5-, CD16+, CD56-, CD57-, CD25+, CD30+, CD103 (alphaEbeta7)+, bcl-2 protein+, CD95+, CD95 ligand(L)-. CD95L, however, was detected in histiocytic and fibroblastoid by stander cells. The lymphomas expressed granzyme B, perforin, and the TIA-1 antigen in various combinations. All three cases had a very unfavorable clinical course characterized by local recurrence and/or dissemination to other epithelial sites, leading to death within 6-12 months after the initial diagnosis despite surgery and aggressive antineoplastic treatment. These data suggest a novel variant of peripheral T-cell lymphoma operationally characterized as primary gastric, apoptosis-rich, CD103+, EBV-, T-cell lymphoma co-expressing CD4, CD8, CD16 and cytotoxic molecules. PMID- 10834540 TI - Mycobacterial DNA in recurrent sarcoidosis in the transplanted lung--a PCR-based study on four cases. AB - Sarcoidosis is a systemic granulomatous inflammation, which may be caused by mycobacteria other than M. tuberculosis complex (MOTT) in one-third of cases. A few cases of recurrent sarcoidosis in the transplanted lung have been reported. However, mycobacteria have been excluded by acid-fast stains only. We investigated four cases of recurrent sarcoidosis in lung transplant patients. Using PCR for the insertion sequence 6110 of Mycobacterium tuberculosis complex and a second PCR for the mycobacterial chaperonin (65-kDa antigen coding sequence), we looked for mycobacterial DNA. In three cases sequence analysis was also performed. One patient was negative for mycobacterial DNA in explanted, but positive for M. tuberculosis DNA in transplanted lung, qualifying this case as M. tuberculosis infection in the transplant. Three patients were negative for M. tuberculosis DNA, but were positive for MOTT-DNA in both explanted and transplanted lungs. In these three patients sequence identity of the amplified sequences before and after transplantation was proven, which rules out mycobacteriosis. Recurrent sarcoidosis does occur, but can only be proven by the exclusion of mycobacterial DNA. In cases of recurrent MOTT-DNA-positive sarcoidosis the diagnosis cannot be confirmed except by proof of sequence identity. Probably MOTT-DNA-positive sarcoidosis is more likely to recur in a transplanted lung. PMID- 10834541 TI - Overexpression of cyclin D1 in nonmelanocytic skin cancer. AB - Although the overexpression of cyclin D1 has been believed to play important roles in neoplastic transformation of some tumors, little is known about the function of cyclin D1 protein in carcinogenesis in human skin. A total of 307 patients with nonmelanocytic skin cancer, being 46 with Bowen's disease (BOD), 134 with squamous cell carcinoma (SCC) and 127 with basal cell carcinoma (BCC), were investigated immunohistochemically using monoclonal antibody to cyclin D1 by the LSAB method, to assess the expression of cyclin D1 in skin cancer including its precursors. The positive rates of cyclin D1 immunostaining in BOD, SCC and BCC were 63.0%, 69.4% and 54.3%, respectively. The positive rates in dysplasia adjoining BOD, SCC and BCC were 43.6%, 67.9% and 59.8%, respectively. In morphologically normal skin, however, only 2 cases, 1 of SCC and 1 of BCC, exhibited positive staining. These findings suggested that overexpression of cyclin D1 is an early event in dysplastic lesions of skin. Overexpression of cyclin D1 was related to sun exposure, especially in dysplasia of SCC. The score for cyclin D1 expression in dysplasia of BCC was correlated with age. Expression of cyclin D1 markedly increased from normal skin through dysplasia to BOD, but was not significantly related to the degree of SCC differentiation. These findings demonstrate that the effect of cyclin D1 overexpression is restricted to proliferation of cells, so that they gain a growth advantage, but their differentiation is not increased. Comparison with the results for p53 protein expression in these tumors, a significant correlation with cyclin D1 expression was found in dysplasia in BOD and SCC, and in patients with BCC who were less than 74 years old. These findings suggested the hypothesis that prior aberrant p53 expression may affect or regulate the overexpression of cyclin D1. PMID- 10834542 TI - Early cellular and ultrastructural response of the mouse urinary bladder urothelium to ischemia. AB - An experimental ischemic model of mouse urinary bladder was developed to study urothelium permeability and changes in cell ultrastructure. The bladder permeability barrier response to experimental ischemia (30-120 min) was investigated by means of indigo carmine dye, trypan blue and lanthanum nitrate tracer, which were used as quantitative and qualitative indicators of urothelial integrity. Changes to the urothelium were studied by light microscopy, and by scanning and transmission electron microscopy. It was established that ischemia primarily induces breakdown of the blood-urine permeability barrier by disruption of the tight junctions. It causes focal interruption of the contacts between the cells, which is followed by detachment and desquamation of viable urothelial cells. Urothelial damage occurs as funnel-shaped wounds, which can extend into the lamina propria. They are proportional to the duration of ischemia and to the extent of reperfusion induced. Desquamated cells in the bladder lumen, when exposed to hypertonic and toxic urine, gradually become irreversibly changed. PMID- 10834543 TI - An intrapulmonary teratoma associated with bronchiectasia containing various kinds of primordium: a case report and review of the literature. AB - An intrapulmonary teratoma (IPT), multiloculated and bronchiectatic, with two polyps inside a 23-year-old man is reported. The IPT, a very rare benign cystic lesion, was communicating with segmental bronchus and was removed by a segmental resection from the upper lobe of the left lung. The teratoma contained various kinds of primordial derivatives, such as mesoderm, ectoderm, and endoderm. Though 65 cases of IPT have been reported in the literature (1839-1996), in the present case there were over 15 germ derivatives, the largest number reported to date. The tumor contained thymic tissue, apart from mediastinum, which may be significant in relation to the pathogenesis of IPT. Clinical manifestations, age, and gender distributions and the kind of germ cell derivatives are discussed. PMID- 10834544 TI - Multifocal micronodular pneumocyte hyperplasia in a postmenopausal woman with tuberous sclerosis. AB - We report a peculiar case of multifocal micronodular pneumocyte hyperplasia (MMPH) without association of pulmonary lymphangioleiomyomatosis (LAM) in a 56 year-old postmenopausal woman with tuberous sclerosis. This case is surmised to be a forme fruste of tuberous sclerosis. Computed tomography demonstrated multiple micronodules, measuring up to 5 mm in size, present in the bilateral lung fields, but no cystic changes. A proliferation of pleomorphic type-II pneumocytes lining the thickened alveolar septa in an adenomatoid pattern, with an associated increase in alveolar macrophages, was observed without typical nuclear atypia. In fully developed lesions, the ingrowth of more proliferating type-II pneumocytes into the thickened alveolar septa and macrophages filling the alveolar lumens were characteristic findings. Proliferation of immature smooth muscle cells suggesting LAM was not observed. Positive immunohistochemical stains for cytokeratin, epithelial membrane antigen, and surfactant apoproteins A and B, and negative staining for HMB45, alpha-1 smooth muscle actin, desmin, and carcinoembryonic antigen confirmed the characteristics of alveolar lining cells in each MMPH lesion. MMPH associated with tuberous sclerosis in the postmenopausal woman appears to be similar to that described in premenopausal women. The present case is familial rather than sporadic and suggests no relationship between the development of MMPH and the underlying hormonal state. PMID- 10834545 TI - Nasal NK/T cell lymphoma presenting as transverse myelopathy. AB - A case of nasal NK/T cell lymphoma with central nervous system (CNS) involvement is reported. A 56-year-old man presented with eyelid edema and transverse myelopathy. Cerebrospinal fluid examination revealed atypical lymphoid cells with azurophilic granules, which were positive for CD2, CD8, and CD56, and negative for CD3 and CD5 by flow cytometry. Because a tumor mass was found involving the ethmoid and maxillary sinuses, CNS involvement was considered to have resulted from local invasion by the nasal lymphoma. In spite of intensive chemotherapy including intrathecal infusion, the patient died 6 months after the initial diagnosis. Autopsy revealed that lymphoma cells were positive for cytotoxic molecules, granzyme B and TIA-1, and EB virus-encoded RNA-1 (EBER-1), and they showed no rearrangement of TCR-beta, -gamma, or -delta genes, suggesting an NK cell origin of the lymphoma cells. They showed an angiocentric and angiodestructive pattern in the subarachnoid space, focally extending to the cerebral cortex and cranial and spinal nerve roots. Marked demyelination was found in the lateral and posterior funiculi of the spinal cord. Thus, the pathogenesis of this spinal demyelination might be attributed to ischemia secondary to angiocentric and angiodestructive infiltration by lymphoma cells. PMID- 10834546 TI - Intravascular papillary endothelial hyperplasia in the kidney of a child. AB - Intravascular papillary endothelial hyperplasia (IPEH) is a benign vascular lesion which is thought to represent an unusual form of organizing thrombus. A case of IPEH in the kidney of a 7-year-old girl is described. She suffered from intermittent flank pain and gross hematuria for 6 months. On radiological examinations, well-defined hypoechoic lesions were identified in the medullary portion of the left kidney. A well-demarcated, sponge-like mass was noted on gross examination. It was an intravascular mass lined by a fibrous capsule of various thicknesses. It was characterized by papillary fronds lined with benign endothelial cells. This is the first description of a renal IPEH in a child. PMID- 10834547 TI - Septal replacement for severe anterior septal deviation. AB - PURPOSE: To introduce a novel surgical technique for treatment of severe anterior septal deviation. MATERIALS AND METHODS: Retrospective case series set in a tertiary-care hospital. Ten patients were treated over an 18-month period. External septorhinoplasty and septal replacement for severe anterior septal deviation. Postoperative clinical nasal examination and photographic comparison. RESULTS: Ten patients were treated with septal replacement for severe anterior septal deviation over an 18-month period. All patients were followed for a minimum of 1 year. No postoperative complications were seen. All patients reported a subjective improvement in their nasal airway and nasal appearance. This was confirmed with clinical examination and photographic comparison. CONCLUSION: Septal replacement for severe anterior septal deviation is a viable option for select patients who can not be successfully treated with conventional techniques. PMID- 10834548 TI - Effects of platelet-derived growth factor-AA on the healing process of tympanic membrane perforation. AB - PURPOSE: Platelet-derived growth factor basic 30-kD disulfide-bonded dimer of A and B chains (PDGF-AA, PDGF AB, PDGF-BB) and a cytokine, promoting wound healing by its mitogenicity for fibroblast and by stimulating the production of fibronectin and hyaluronic acid. This article investigates the effect of PDGF on the healing process of tympanic membrane (TM) perforation. MATERIALS AND METHODS: The pars tensa of the posterior aspect of the TM of rats was excised and treated with 2 microg of PDGF-AA or placebo. The animals were killed at 3, 5, 7, 9, 11, 15, and 28 days after operation. The healing process of TM perforation was observed with a telescope and light microscope. The temporal bones were also immunohistochemically examined for PDGF-alpha receptor (PDGF-R(alpha)) and fibronectin. RESULTS: All PDGF-AA-treated TM were completely closed by 5 days after surgery, whereas some of the placebo-treated TM were not closed at 15 postoperative days. PDGF-AA induced the most prominent proliferation of the connective tissue by 9 postoperative days, after which the growth of the connective tissue decreased. By the 4th postoperative week, the PDGF-treated TM were slightly thicker than normal TM. An intense expression of fibronectin was detected in the connective tissue layer of the TM that were treated with PDGF-AA. PDGF-R(alpha) was expressed in the epithelial layer of both the PDGF-treated and control TM. CONCLUSION: These results show that PDGF-AA speeds up the healing process of TM defect, improves the rate of healing, and prevents atrophic changes in the healed TM by promoting the connective tissue growth. The use of PDGF-AA can be an effective alternative to surgery for managing TM perforations. PMID- 10834549 TI - Petrous high jugular bulb: a histological study. AB - PURPOSE: A high jugular bulb (JB) is thought to affect structures of the inner ear and possibly cause symptoms there, but clear histological findings of an anatomical relationship between a high JB and the inner ear have not yet been described. MATERIALS AND METHODS: We surveyed horizontal sections of 1,591 temporal bones from the collection of the Otopathology Laboratory at the University of Minnesota in Minneapolis, Minnesota, defining a high JB as a JB extending above the inferior margin of the basal cochlear turn. RESULTS: In 65 specimens (16%), we found a high JB with its vascular wall obviously thinner than that of a low JB. Bony resorption was occasionally observed around high JBs. Sixteen specimens showed a bony deshiscence between the JB and the endolymphatic sac. Clinical charts showed no obvious symptoms associated with a high JB. CONCLUSIONS: Our findings suggest that the JB may have potential to expand upward postnatally. Although our study confirmed occasional bony dehiscence between the JB and the endolymphatic sac, JBs with this involvement may have only a minor effect on function in the inner ear. PMID- 10834550 TI - Deep neck infections in diabetic patients. AB - PURPOSE: Deep neck infections may be lethal if life-threatening complications occur, especially in immunocompromised hosts such as diabetic patients. This study was undertaken to better define the clinical features and prognosis of deep neck infections in the diabetic patients with special emphasis on the use of empirical antibiotics and the role and timing of surgical management. PATIENTS AND METHODS: A retrospective analysis of 105 consecutive patients treated at a single institution during a 9-year period was conducted. Of these, 30 patients also presented with diabetes mellitus. RESULTS: Compared with the nondiabetic patients, the unique features of deep neck infections in diabetic patients were as follows: (1) older age, (2) tendency of unclear infection source, (3) tendency to involve multiple spaces, (4) required more aggressive surgical intervention, (5) prolonged hospitalization, and (6) higher complication rate. The differences were statistically significant (P< .05). There were no significant differences in the complete blood count/ differential count positive findings and fever between the 2 groups (P> .05). Bacterial cultures showed that Klebsilla pneumoniae was the most common causative pathogen in diabetic patients. CONCLUSIONS: In deep neck infection patients with diabetes mellitus, the clinical course is more severe and there is a poorer prognosis. Thus, in treating them, we should keep close observation, appropriately control the diabetes, detect the life threatening complications early, perform aggressive surgical treatment if fluctuation or complication occurs, and take into account the preponderance of K pneumoniae when administering the empirical antibiotics. PMID- 10834551 TI - Radiation therapy in inverted papillomas of the nasal cavity and paranasal sinuses. AB - PURPOSE: Between December 1969 and September 1989, a total of 10 patients with advanced and/or recurrent inverted or cylindrical cell papillomas were treated with irradiation at the University of Florida in Gainesville. MATERIALS AND METHODS: Nine of 10 patients had 1 or more recurrences before they received radiation therapy. Three patients were treated with irradiation alone, and 7 patients received surgery and irradiation (preoperatively in 1 patient and postoperatively in 6 patients). Eight patients had inverted papillomas (3 with concomitant squamous cell carcinoma), and 2 patients had cylindrical cell papillomas. RESULTS: Local recurrence developed in 4 patients at 1.5, 6.5, 12, and 13 years after treatment. No evidence of recurrence was observed in 6 patients at 7, 8.5, 8.5, 9, 9, and 20.5 years after treatment. Four patients died of intercurrent disease. No patient developed a malignant transformation. Significant complications of treatment included, in 1 patient, an area of bone exposure in the orbit that necessitated debridement. CONCLUSION: Surgery is the primary treatment of this entity. Radiation therapy should be considered in patients with incompletely resectable lesions, multiply recurrent tumors, and tumors associated with malignancy. PMID- 10834552 TI - Computerized tomographic alignment of silastic implant in type 1 thyroplasty. AB - PURPOSE: We designed a computerized tomography (CT)-based silastic implant preparation method that enabled custom fit to the individual size of the patient's larynx for medialization laryngoplasty. MATERIALS AND METHODS: Three women with unilateral vocal cord paralysis underwent type I thyroplasty operation. The individual size of the patient's larynx was determined by preoperative measurements on CT scan and the implant was prepared accordingly. The implant was then inserted through a rectangular window at the level of vocal cords which had been outlined according to CT findings. RESULTS: Three patients, who were age 41, 25, and 37 years, underwent medialization laryngoplasty by this technique. They were followed up for 37, 16, and 4 months, respectively. There was not any rejection reaction, and satisfactory functional results with 10, 7, and 9 seconds of phonation duration have been achieved, respectively. CONCLUSION: In this technique, the desired medialization of the paralyzed vocal cord was accomplished by the first insertion of the implant. Thus, the duration of the operation and the vocal cord edema aroused by manipulation of the inner perichondrium and internal laryngeal structures were reduced. PMID- 10834553 TI - Myxomas of the head and neck. AB - PURPOSE: Myxomas are rare, locally infiltrative, benign, connective tissue tumors that are found in bone and somatic soft tissues. This article reports our experience with head and neck myxomas and provides a comprehensive literature review. MATERIALS AND METHODS: Retrospective record review of head and neck myxoma cases seen at 2 tertiary referral centers in Houston, Texas, from 1970 to 1994. Comprehensive literature review and compilation of all myxomas reported in the head and neck. RESULTS: We identified 7 cases of true head and neck myxoma treated in our centers and identified 169 cases reported in the literature. Most often, myxomas originated in bone (mandible or maxilla) and were most commonly found in adults. All tumors except 1 were treated surgically. Recurrence rates were 6% for local or wide excision and 28% for more conservative surgery such as enucleation or curretage. CONCLUSIONS: Myxomas of the head and neck should be treated with complete excision of the tumor with clear margins. PMID- 10834554 TI - Penetrating injuries of the visceral compartment of the neck. AB - Penetrating injuries of the visceral compartment of the neck are uncommon but potentially life threatening. A retrospective review of patients who sustained penetrating trauma to the laryngotracheal complex was conducted at the Level I trauma center of the University of Louisville Hospital in Kentucky. Sixteen patients were identified and their records reviewed for type of injuries, treatment, complications, and 1-year follow-up. The majority of patients were men who sustained injuries that were violent in nature. Zone II of the anterior neck was the most commonly injured area, with the trachea (69%), esophagus (38%), and larynx (31 %) the most commonly injured structures. Although 31% underwent angiograms, only 13% showed vascular injuries. Eighty-one percent of the patients had injuries involving more than 1 major structure of the neck. Neck exploration was performed in 81% of the patients and tracheotomies in 75% as well as repair of the trachea (50%), larynx (31%), and esophagus (38%). There is significant mortality associated with these injuries (13% in our study), and many of the patients have long-term sequelae such as dysphagia, hoarseness, and prolonged tracheotomy. PMID- 10834555 TI - Cricothyroidotomy: when, why, and why not? PMID- 10834556 TI - The use of unilateral deep plane neck lifting to improve the aesthetic appearance of the neck dissection deformity. AB - The primary concerns of head and neck cancer surgeons are, and should remain, the complete extirpation of tumor and the prevention of tumor recurrence. In recent years, numerous advances have been made in the reconstruction of this patient population, significantly improving their functional and aesthetic outcomes. In this article, with an illustrative case example, we present our technique of unilateral deep plane neck lift that may be considered when one is attempting to achieve better symmetry in patients after radical neck dissection. PMID- 10834557 TI - Desmoid-type infantile fibromatosis in the mandible: a case report. AB - A 3-year-old boy with desmoid-type infantile fibromatosis arose in the mandible was reported. He was referred to our hospital because of suspected malignant bone tumor of the mandible. Histological examination of an open biopsy specimen was performed followed by tumor resection with marginal mandibulectomy and reconstruction by iliac bone grafting, which caused no functional complications nor mandibular deformity. To treat tumors in the facial skeleton, the surgical procedure should be planned based on the histological diagnosis in order to determine the proper area of resection to prevent functional or cosmetic complications. Especially in children, attention should be taken for benign but clinically resembling malignant rare diseases such as desmoid-type infantile fibromatosis. PMID- 10834559 TI - Congenital tracheal agenesis: a postmortem contrast radiographic study. PMID- 10834558 TI - Ewing's sarcoma of the facial zygomatic area bones in a young child: a case presentation. AB - We present the case of a young patient with Ewing's sarcoma of the facial zygomatic area bones. This type of tumor in a very young child is a rare event and poses significant diagnostic and therapeutic challenges for the attending physician. In this case, the diagnosis was made by a computed tomography scan with subsequent histological confirmation. The differential diagnoses and therapeutic options are discussed. PMID- 10834560 TI - Presentation and treatment of a posttraumatic pseudomeningocele of the superior orbit. PMID- 10834561 TI - Effect of tropically adapted sire breeds on preweaning growth of F1 Angus calves and reproductive performance of their Angus dams. AB - The objective of this study was to determine the effect of tropically adapted sire breeds on preweaning growth performance of F1 calves and on reproductive performance of their Angus dams. Angus (A) cows were bred in two consecutive years (1992 and 1993) by AI using semen from Brahman (B; Bos indicus; n = 10), Senepol (S; Bos taurus; n = 10), and Tuli (T; Sanga; n = 9) bulls. A total of 82 B x A, 85 S x A, and 91 T x A calves were born. The statistical model included the fixed effects of year, sire breed, calf sex, sire breed x calf sex, and cow parity and the random effect of sire within sire breed. Birth weight, weaning weight, 205-d adjusted weaning weight, ADG from birth to weaning, and hip height at weaning were greater (P < .001) for B x A calves than for S x A or T x A calves. Greater differences were detected between sexes for B x A than for S x A and T x A (for all traits sire breed x calf sex, P < .05). Sire breed affected (P < .01) the percentage of unassisted calvings (B x A, 87%; S x A, 98%; and T x A, 100%) and tended (P < .10) to affect the percentage of calves that survived until weaning (B x A, 90%; S x A, 94%; and T x A, 98%). Sire breed of calf did not affect (P > .10) length of gestation, and sire breed did not affect the interval from calving to first observed estrus or pregnancy in Angus dams. These results demonstrate that preweaning growth performance of B x A calves was greater than that of either S x A or T x A calves. However, use of Brahman sires on Angus dams led to calving problems and tended to reduce the percentage of calves that survived until weaning. Thus, heavier weaning weights of B x A calves would be an advantage for cow-calf producers marketing calves, but heavier birth weights and calving difficulty attributed to Brahman sires would be a disadvantage. PMID- 10834563 TI - Effect of gilt development diet on the reproductive performance of primiparous sows. AB - The objective of this study was to evaluate the effect of development diet on first-parity reproductive performance across different genetic types of females. Gilts (n = 708) 8 to 15 d of age from five genetic lines were assembled using a segregated early weaning protocol. Genetic types represented industry variation for reproductive capacity and lean growth potential. Sampling procedures were not designed to evaluate performance differences among the genetic lines. When the gilts weighed approximately 20 kg, they were moved from the nursery facilities to a slotted-floor, environmentally controlled facility, and seven to eight animals within a genetic type were penned together. When the gilts weighed approximately 40 kg, they were moved to a modified open-front facility. Nineteen gilts were allotted to each pen (.92 m2 per pig). Gilts were assigned to one of three development diets at 120 d of age. Diet 1 (high energy, 18% CP) and Diet 2 (high energy, 13% CP) were provided for ad libitum consumption to the assigned gilts until they weighed approximately 113 kg. Gilts receiving Diet 3 (23% CP) were fed 1.8 kg/d from 82 kg until they reached 180 d of age (approximately 100 kg). Gilts were fed 2 kg daily of a gestation diet from 180 d to 200 d of age and 2.7 kg daily from 200 d until mating. To stimulate the estrus cycle, gilts were commingled and exposed to vasectomized boars beginning at 180 d of age. Gilts that were in estrus and 210 d of age or older were artificially inseminated with commercial semen. Gilts not detected in estrus within the first 50 d of observation were injected with PG600 and estrus detection continued for 30 additional days. Of the 657 gilts entering breeding pens, 422 farrowed. Bred gilts were distributed to 10 cooperator facilities before farrowing. Mixed model procedures were used to analyze the data. Significant (P < .05) genetic type x gilt development diet interactions were found for number of pigs born, number of pigs born alive, total litter birth weight, and litter birth weight of pigs born alive. Significant interactions consistently involved one genetic line and gilt development Diets 1 and 2. Gilts from this genetic line-diet subclass had poorer farrowing performance (P < .05) than gilts from the same line fed development Diet 3. Only two other significant genetic line x gilt development diet interactions were found. Gilt development diet had little influence on first parity reproductive performance. PMID- 10834562 TI - Body weight and tissue gain in lambs fed an all-concentrate diet and implanted with trenbolone acetate or grazed on alfalfa. AB - Targhee x Hampshire lambs (average BW 23 +/- 1 kg) were used in two experiments to determine the effects of finishing on concentrate with an anabolic implant or forage grazing after concentrate feeding on growth, organ and viscera weights, and carcass tissue accretion. In Exp. 1 and 2 lambs were penned by sex and assigned for slaughter at initial (23 kg), intermediate (37 kg), or end BW (ewes, 47.7; wethers 50.4 kg). From 23 to 37 kg BW, lambs were fed all-concentrate diets in drylot (DL) or grazed on alfalfa (ALF). Experiment 1 was a 2 x 2 factorial with 28 lambs; factors were wether vs ewe lambs and unimplanted vs DL implanted with trenbolone acetate-estradiol benzoate. There were no differences in organ and viscera weights due to implant status. However, ADG (P < .03) and lean gain (P < .02) were greater for implanted than for unimplanted wethers (507 vs 357 g and 1,314 vs 656 g, respectively). Ewes did not respond to the implant. Fat accretion was not affected by implantation. Experiment 2 was a 2 x 3 factorial with 42 lambs; factors were wether vs ewe lambs and drylot during growing and finishing phases (DL-DL) vs drylot during growing and alfalfa grazing during finishing (DL-ALF) vs alfalfa grazing during growing and finishing phases (ALF ALF). In Exp. 2, ADG of DL-DL lambs was greater (P < .01) than ADG of DL-ALF or ALF-ALF lambs. Lambs on ALF-ALF had smaller (P < .05) livers and rumen/reticulum weights but heavier (P < .04) kidney, omasum, small and large intestine, and cecum weights than those on DL. In Exp. 2, DL-ALF and ALF-ALF lambs had overall hindsaddle lean gain equal to those on DL-DL with less mesenteric fat and 100 g less separable fat. Finishing lambs on alfalfa reduced fat accretion without decreasing lean accretion, whereas trenbolone acetate implants for lambs fed concentrate increased BW gain and lean accretion without affecting fat accretion. PMID- 10834564 TI - Preweaning growth curves in Brown Swiss and Pirenaica calves with emphasis on individual variability. AB - A quadratic polynomial model with random regression coefficients was used to describe preweaning growth curves of two beef cattle breeds widely used in the Spanish Pyrenees, according to genotype and season of birth effects. In addition, parameters of individual variability that can be used in a stochastic model were obtained. Data recorded indoors from birth to weaning of 217 Brown Swiss calves (3,509 observations) born either in spring or autumn (BS-S, BS-A) and 101 spring born Pirenaica calves (PI-S, 967 observations) were analyzed. A quadratic model accurately fitted the preweaning weights (R2 = .99). Use of random regression coefficients improved the weaning weight adjustment; the residual variance of the model with intercept and linear random coefficients (9.61 kg2) was smaller than that of the model without them (130.03 kg2). Brown Swiss-S and PI-S calves had similar birth weight (40.9 +/- .96 vs 39.4 +/- .73 kg), but BS-S calves achieved significantly higher weaning weights at 150 d of age (175.2 +/- 2.45 vs 158.4 +/- 3.17 kg). Preweaning growth patterns were different for each season of birth, but there were no differences in weaning weight at 150 d of age (172.9 +/- 2.01 BS-A vs 175.2 +/- 2.45 BS-S). Standardization of weaning weights using a linear approximation could lead to biases, especially when comparing animals from the two calving seasons. The estimate of variances of random parameters should be done within breed and season of birth in order to take into account heteroscedasticity. The variances for BS-A, BS-S, and PI-S were 39.9, 57.6, and 32.2 kg2 for the intercept, respectively, and .0159, .0141, and .0205 kg2 for the linear coefficient. Covariance between the intercept and the linear coefficient (.34 kg2) was only statistically significant in the case of BS-S. The individual variance of weight at 150 d was 424.7 kg2 and 526.7 kg2 for BS-S and PI-S, respectively, almost 65% of the observed variance of weaning weight. PMID- 10834565 TI - Estimates of genetic parameters for hip and elbow dysplasia in Finnish Rottweilers. AB - Data from 2,764 Rottweiler dogs born from 1987 to 1996 were analyzed with a Restricted Maximum Likelihood procedure using a mixed linear animal model to obtain variance component estimates for hip and elbow dysplasia. The data included 2,764 hip dysplasia and 2,278 elbow dysplasia records. Hip joints were scored as normal (0), borderline (1), slight (2), moderate (3), and severe (4, 4.5, and 5) hip dysplasia. Elbow joints were graded normal or borderline (0), slight (1), moderate (2), and severe (3) elbow dysplasia. The mean for the hip scores was 1.07 and for the elbow scores .60. Environmental effects influencing hip dysplasia were age, birth year, birth year x season interaction, and experience of the veterinarian responsible for x-raying the dog. For elbow dysplasia, statistically significant effects were age, birth year, sex of the dog, and panelist responsible for each screening. Estimates of heritability for hip and elbow dysplasia were .58 +/- .04 and .31 +/- .04, respectively, with a genetic correlation of .37 +/- .08 between the traits. Genetic improvement of almost one genetic standard deviation was observed in both traits during the 10 yr covered by the data. PMID- 10834566 TI - Duration of the seasonal anestrus in sheep selected for fertility in a fall lambing system. AB - Crossbred ewes (1/2 Dorset, 1/4 Rambouillet, and 1/4 Finnsheep) from a flock being selected for spring fertility, defined as ability to lamb following ram exposure in May and June in Virginia (37 degrees N latitude), were used to study the duration of the seasonal anestrus. In the first 3 yr of the study (1992, 1993, and 1995), mature ewes were divergently selected based on EBV for fertility, and the duration of anestrus was measured by continuously exposing the ewes to vasectomized rams equipped with marking harnesses from mid-January until approximately August 1. Only ewes that had lambed in the previous fall were used to ensure that ewes were in a comparable physiological state, and the same rams remained with the ewes in each year to avoid induction of estrus by introduction of novel rams. The duration of anestrus in high-fertility ewes (n = 26; mean fertility EBV of 12.6%) was 28.4 d, which was significantly less than the 70.2 d of anestrus observed for low-fertility ewes (n = 15; mean fertility EBV of .3%). Five high-fertility ewes did not exhibit a period of anestrus. The regression of number of days of anestrus on fertility EBV was -2.15 +/- .72 d/%. In yr 4 (1997), 11 high-fertility and two low-fertility ewes were evaluated. None of these ewes exhibited a clear seasonal anestrus; six unequivocally cycled continuously. Between January 23 and July 31, the mean duration of anestrus for these ewes was only approximately 11 d. The duration of anestrus for high fertility ewes seems to be the shortest reported for temperate sheep breeds. PMID- 10834567 TI - Genetic correlations for daily gain between ram and ewe lambs fed in feedlot conditions and ram lambs fed in Pinpointer units. AB - When performance is recorded in automated facilities that measure feed intake of individual lambs that are penned in a group, such as Pinpointer units, a legitimate question is the degree to which daily gain is genetically correlated with daily gain achieved under feedlot conditions. Lambs were from a composite population (1/2 Columbia, 1/4 Suffolk, and 1/4 Hampshire germplasm) and of the F2 or more advanced generations. Data were daily gains of 1,101 rams (PR) fed in Pinpointer units (11 to 17 wk of age) and 2,021 rams (FR) and 3,513 ewes (FE) fed under feedlot conditions (9- or 10-wk period starting at 9 wk of age). The FR and FE lambs were born from 1983 through 1995, whereas the PR lambs were born from 1986 through 1995. Measurements of daily gain in PR, FR, and FE lambs were considered to represent three correlated traits. Unadjusted means were .411, .406, and .326 kg/d for PR, FR, and FE, respectively. Random effects in the model were animal direct genetic, maternal genetic, and maternal permanent environmental. Fixed effects were associated with age of dam (1 to 6 yr), type of rearing (1 to 4), and contemporary group (test date). Variances due to maternal genetic effects with single-trait analyses were near zero, so those effects were eliminated from the three-trait analysis although a random uncorrelated effect due to dam was included in the model. Estimates of heritability were .22, .14, and .23 for PR, FR, and FE, respectively, with fractions of variance due to dam effects ranging from .02 to .05. Estimates of genetic correlations were .86 for PR with FR, .83 for PR with FE, and 1.00 for FR with FE. Estimated phenotypic variances were similar for PR and FR, but one-third less for FE. The similarity of heritability estimates and estimates of genetic correlations all exceeding .83 suggest that daily gain of rams fed in Pinpointer units will reflect genetic expression for daily gain in both ram and ewe lambs fed under feedlot conditions. PMID- 10834568 TI - Simulation of cow-calf production systems in a range environment: I. Model development. AB - A mathematical computer model of beef cattle production systems was developed at Montana State University. The objective of this report was to describe the rationale and procedures used to simulate animal and system performance. The model was designed to simulate the dynamic relationships among cattle genotype, physiological state, forage quality, and management in range environments. Forage intake, energy and protein metabolism, growth, reproduction, lactation, and changes in chemical body composition are simulated for individual animals over complete life cycles. Expenses driven by animal performance, management decisions, and land resources are tabulated. Several biological and economic measures of system performance can be computed, including ratios of inputs (e.g., DM, CP, ME, dollars) to outputs (e.g., weight, lean), break even prices, and annual gross margin per cow or ranch. Primary uses of the model include the evaluation of system responses to changes in breeding strategies and management in range production/marketing systems. PMID- 10834569 TI - Simulation of cow-calf production systems in a range environment: II. Model evaluation. AB - Development of a general beef cattle simulation model was described in a companion paper. The objectives of this research were to demonstrate and evaluate the model's performance. Four experiments were conducted to demonstrate 1) life cycle weight and body condition changes for different genotypes raised in a northern range environment; 2) responses in forage intake and weight to changes in forage quality, protein supplementation, and cow physiological state; 3) responses in reproduction, weight, body condition, and calf growth to differences in pre- and postpartum nutrition; and 4) differences in enterprise efficiency and profit for different genotypes and mating systems. Results indicated that the model performs acceptably for the types of situations for which it was designed and highlighted areas of animal science where more information is needed in order to better understand and manipulate beef cattle systems. Computer simulation models are useful tools to facilitate the integration of scientific concepts and to help scientists, teachers, and producers better understand the complex production systems that they study and manage. In addition to addressing enterprise-level problems that are beyond the scope of traditional experimentation, modeling and simulation help identify research needs and foster exchange among disciplines. PMID- 10834570 TI - Parameter estimates for genetic effects on carcass traits of Korean native cattle. AB - Data (n = 1,746) collected from 1985 through 1995 on Korean Native Cattle by the National Livestock Research Institute of Korea were used to estimate genetic parameters for marbling score, dressing percentage, and longissimus muscle area, with backfat thickness, slaughter age, or slaughter weight as covariates. Estimates were obtained with REML. Model 1 included animal genetic and residual random effects. Model 2 was extended to include an uncorrelated random effect of the dam. Model 3 was based on Model 1 but also included sire x region x year season interaction effects. Model 4 combined Models 2 and 3. All models included fixed effects for region x year-season and age of dam x sex combinations. From single-trait analyses, estimates of heritability with covariates to adjust for backfat thickness, slaughter age, and slaughter weight from Model 4 were, respectively, .10, .08, and .01 for marbling score; .09, .12, and .16 for dressing percentage; and .18, .17, and .24 for longissimus muscle area. From three-trait analyses, estimates of genetic correlations between marbling score and dressing percentage, marbling score and longissimus muscle area, and dressing percentage and longissimus muscle area were, respectively, -.99, .20, and -.11 with backfat thickness as covariate; -.88, .47, and .01 with slaughter age as covariate; and -.03, .39, and .91 with slaughter weight as covariate. Results of this study suggest that choice of covariate (backfat thickness, slaughter age, or slaughter weight) for the model seems to be important for carcass traits for Korean Native Cattle. Including sire x region x year-season interaction effects in the model for marbling score and dressing percentage may be important because whether sire x region x year-season interaction effects were in the model affected estimates of other variance components for the three carcass traits. Whether the maternal effect was in the model had little effect on estimates of other parameters. With backfat thickness and slaughter age end points, selection for increasing marbling score would be expected to result in decreasing dressing percentage for Korean Native Cattle. With slaughter weight as a covariate for end point, increased longissimus muscle area would be associated with increased dressing percentage, and increased marbling score would be related to increased longissimus muscle area. The differences in estimates associated with choice of end point, however, need further study. PMID- 10834571 TI - Impacts of an endophyte-infected fescue seed diet on traits of mouse lines divergently selected for response to that same diet. AB - In previous work, mouse lines were selected for eight generations for resistance (R) or susceptibility (S) to endophyte-infected fescue toxicosis using depression in postweaning gain caused by a toxin-containing diet as the selection criterion. Characterizing biological changes associated with resistance or susceptibility in those mice might suggest genetic or therapeutic approaches to alleviate fescue toxicosis in cattle. The first objective of the current experiment was to determine whether the toxin-containing diet depressed reproduction and mature size more severely in S than in R mice. The second was to investigate line and diet effects on hepatic glutathione-S-epoxytransferase (GST) and uridine diphosphate glucuronosyl-transferase (UDPGT) activities and to relate enzyme activities to reproduction within line by diet groups. Twenty-eight pairs per line (S or R) x diet (toxin-containing [+] or toxin-absent [-]) group cohabitated for 36 wk. The + diet depressed the number of pups born and weaned and litter weight weaned (P < .01) within the first two litters produced. Diet effects were greatest early in the experiment. Percentage changes in reproduction caused by the + diet for R and S pairs, respectively, were -13 and -28 for total pups born, -10 and -25 for total pups weaned, -13 and -14 for total litters produced, and 30 and -42 for total litter weight weaned. The S line mice were heavier than R line mice on both diets, but the + diet had a larger depressing effect on mature size of S line than of R line males (line x diet interaction, P = .09) and females (interaction not significant). Averaged across diets, GST activity was higher in R than in S dams (P = .05) at 44 wk of age but was not affected by diet or line x diet. Activity of GST was correlated with number of pups born (-.50), number of litters produced (-.44), and survival percentage (.40) within the R- group; in the R+ group, GST activity was correlated only with survival percentage (.37). In the S- and S+ groups, GST activity was not correlated with any reproductive trait. Line, diet, and their interaction did not affect UDPGT activity, and UDPGT activity was not correlated with any reproductive trait in any line x diet group. Selected lines differed in response to a toxin-containing diet as measured by its effect on reproduction and mature size. The R and S mice also differed in GST activity, but GST activity was correlated with reproductive traits only in R-line mice. PMID- 10834572 TI - Effects of on-farm diets for organic pig production on performance and carcass quality. AB - An experiment was conducted to evaluate the effects of a restriction to home grown feedstuffs and abstinence from supplementation with synthetic amino acids (AA), as ideal objectives in organic pig production according to the IFOAM standards, on growth performance and carcass characteristics. One hundred individually housed pigs were allocated to four dietary treatments and fed from growing through finishing to compare three organic barley/wheat-based diets with an isocaloric conventional diet supplemented with synthetic AA. Protein sources in the organic treatments were either faba beans, supplemented with potato protein to the same AA level as the control diet, peas and lupines, or faba beans and lupines, both without further supplementation, leading to a lower level of limited AA. Supplementation of organic diets with potato protein resulted in the same performance as supplementing the conventional diet with synthetic AA, although crude protein levels differed markedly. Pigs fed the organic diets without AA supplementation grew more slowly (P < .05) and had a decreased feed intake in the grower period (P < .05) but nearly the same feed efficiency (P > .05) as pigs fed conventional or organic diets with AA supplementation. Carcass characteristics differed in percentage of lean meat and longissimus area, being lower in the treatments without AA supplementation (P < .05). However, the intramuscular fat was higher without AA supplementation (2.9% fat) than with supplementation (1.2% fat) (P < .01). The data show that the exclusion of AA supplementation resulted in a reduction in pig performance but in an increase in intramuscular fat content; the latter is an important aspect of eating quality characteristics. PMID- 10834573 TI - Self-regulation of intake of polyethylene glycol by sheep fed diets varying in tannin concentrations. AB - Tannins occur in many plant species, and they often suppress intake by reducing nutrient availability or by causing malaise. Polyethylene glycol (PEG) binds to tannins and may thereby increase the availability of macronutrients and decrease malaise. Supplemental PEG increases intake of tannin-containing plants by sheep, goats, and cattle. Given the strong response to supplemental PEG, we speculated that animals might self-regulate their intake of PEG when offered foods high in tannins. The objective of the first experiment was to determine if the amount of supplemental PEG (0, 25, 50, 75, or 100 g; molecular weight, 3,350) affected intake by lambs of a food (milo-tannin mix) containing 20% quebracho tannin. There was a linear relationship (Y = 272 + 1.2X; R2 = .86; P = .023) between the amount of supplemental PEG ingested and the subsequent intake of milo-tannin food by lambs. The objective of the second experiment was to determine whether lambs self-regulated intake of PEG when fed a ration that contained 0, 5, 10, 15, or 20% quebracho tannin and whether they adjusted their intake of PEG when tannin was removed from the diet. There was a positive relationship between the amount of PEG ingested and intake of food and tannin (P = .0001). Lambs fed high-tannin diets ate more PEG than controls (P = .03). Lambs fed the 20% tannin diet ate the most PEG, and controls ate the least PEG. Tannin limited intake of the diets, but PEG attenuated the response to a great degree (P = .065). Immediately after tannin was removed from the ration, lambs that formerly had been fed the 20% tannin ration ate more PEG than lambs fed the other rations (P = .0075). Ten of the lambs (5 from the 20% tannin group, 1 from the 15% tannin, and 2 each from the 10 and 5% groups) continued to eat PEG for 7 d after tannin was removed from their ration. When they were tested again 6 wk after the trial and offered tannin free diets, their intake of PEG had decreased. PMID- 10834574 TI - Reducing veal calves' reactivity to people by providing additional human contact. AB - We studied the importance of the stockperson's behavior on veal calf behavior using 22 veal calves housed in individual crates. Eleven calves received minimal contact from the stockperson, and the other 11 calves were stroked and allowed to suck the stockperson's fingers after each meal during the entire fattening period (21 wk). The effects of this additional contact with the stockperson on the calves' responses to people was studied, when in their home environment (crate) or outside their home environment (singly in a novel arena). When tested in their home environment, the calves receiving additional contact withdrew less from the approach of humans (familiar or unfamiliar) (P < .05) compared with control calves. When tested outside the home environment with a human (familiar or unfamiliar) standing motionless, calves that had received additional contact interacted more frequently and for a longer time with the humans and defecated less often compared with control calves (P < .05). In conclusion, being stroked and sucking the stockperson's fingers seemed to be experienced as positive by the calf, because they reduced withdrawal from and increased approaches to familiar and unfamiliar humans in familiar and unfamiliar environments. Such a lower reactivity to people could improve ease of handling, animal performance, and animal welfare. PMID- 10834575 TI - The impact of gentle contacts on ease of handling, welfare, and growth of calves and on quality of veal meat. AB - It has been demonstrated previously that regularly stroking and letting calves suck fingers leads to less avoidance and more approach behavior of the calves toward people. To examine whether these positive contacts affect the welfare and productivity of calves and the quality of veal meat we used 22 veal calves housed in individual crates. Half of them received minimal contact with the stockperson (controls), and the other half were given additional gentle contacts around meals, by stroking the calves and allowing them to suck the stockperson's fingers, during the entire fattening period (21 wk). Welfare was assessed through behavioral reactivity (reactions to handling, to surprise stimuli, and to novelty), neuroendocrine responses to stress (cortisol in response to an ACTH challenge, catecholamine-synthesizing enzymes), and health (number of medical treatments, abomasal lesions). Calf productivity was assessed through growth rates and meat quality through glycolytic potential (an estimator of resting glycogen level in muscle), pH, and color. Calves that received gentle contacts were less agitated (P < .01) and tended to defecate less (P = .08) when handled in a cart on wheels than the control calves, but no treatment effects were found in reactivity to novelty and surprise stimuli, responses to ACTH, and catecholamine synthetic potential. Calves given gentle contacts had fewer abomasal lesions than controls (0/11 vs 4/11, P = .05). The glycolytic potential of the semimembranosus muscle was higher in calves that received gentle contacts than in controls (172.6 vs 154.1 micromol/g, P < .05), but no treatment effects were observed on meat pH, meat color, or growth rates. It is concluded that gentling veal calves reduces their reactions to handling. Gentle contacts reduce the reaction to transport shown by differences in glycolytic potential. In addition, the reduction in reactions to handling and the decreased incidence of abomasal lesions can contribute to an improvement of the calves' welfare. PMID- 10834576 TI - The influence of dexamethasone and insulin on expression of CCAAT/enhancer binding protein isoforms during preadipocyte differentiation in porcine stromal vascular cell cultures: evidence for very early expression of C/EBPalpha. AB - Expression of CAAT/enhancer binding protein (C/EBP) isoforms was examined in primary cultures of adipose tissue stromal vascular (S-V) cells before and during preadipocyte differentiation. Immunocytochemistry showed that the proportions and numbers of C/EBPalpha-, C/EBPbeta-, and C/EBPdelta-reactive cells were maximized after seeding and plating from d 0 to 3 in fetal bovine serum (FBS). However, there were few preadipocytes (AD-3+) and fewer cells with lipid and the number of C/EBPalpha-reactive cells clearly exceeded the number of preadipocytes. Seeding and plating in dexamethasone (DEX) + FBS from d 0 to 3 markedly increased the proportions and numbers of preadipocytes and C/EBPalpha-reactive cells compared to seeding and plating in FBS, d 0 to 3. The number of C/EBPalpha- and C/EBPbeta reactive cells and preadipocyte reactivity for C/EBPbeta decreased with insulin or DEX treatment, d 3 to 6, following FBS, d 0 to 3. However, insulin + DEX treatment, d 3 to 6, maintained the number of C/EBPalpha-reactive cells and either maintained or increased preadipocyte reactivity for C/EBPalpha and C/EBPbeta. DEX and DEX + insulin treatment induced recruitment of a similar number of preadipocytes, but preadipocytes were not reactive for C/EBPalpha and C/EBPbeta in DEX-treated cultures. The number of C/EBPdelta reactive cells did not change from d 3 to 6 and was not influenced by hormone treatment. After DEX + FBS, d 0 to 3, the high numbers of C/EBPalpha-reactive cells and preadipocytes were maintained by insulin treatment alone. Western blot analysis for C/EBPalpha confirmed the immunocytochemical results. Double staining demonstrated that expression of C/EBPalpha protein was maximized before or at the onset of lipid accretion, whereas expression of C/EBPbeta protein was correlated with lipid accretion. These results indicate that coupling or integration of preadipocyte recruitment with C/EBPalpha expression may be a critical step in glucocorticoid induced adipogenesis. PMID- 10834577 TI - Somatotropin and adipose tissue metabolism: substrate and temporal effects. AB - The purpose of these studies was to determine the time course for changes in feed intake, blood metabolites, and lipogenic activity in adipose tissue in response to the initiation of porcine somatotropin (pST) treatment and following withdrawal from treatment in barrows. An initial study was conducted to determine the impact of chronic pST treatment (4 wk of daily injection; 0 vs 4 mg/d) on adipose tissue lipid metabolism in barrows (initial weight 67 kg). Feed efficiency was improved 27%, backfat thickness was decreased 43%, and glucose and lactate oxidation and incorporation into lipid in adipose tissue was reduced 70 to 86% in pST-treated pigs. Palmitate esterification was decreased 44%, whereas palmitate oxidation was unaffected. In vitro metabolism of lactate, glucose, and palmitate in liver slices was not affected by pST treatment. The time-course for changes in intake and adipose tissue metabolism in response to 7 d of pST (0 vs 4 mg/d) treatment and 7 d of withdrawal was examined in subsequent studies in barrows (initial weight 75 kg). Feed intake during pST treatment was significantly (P < .05) less than in control pigs within 24 h of the initiation of treatment and remained low through 3 d after withdrawal. Adipose tissue biopsies were obtained on d 0, 1, 2, 4, and 7 of the treatment phase and on d 2, 4, and 7 after withdrawal from 7 d of treatment. Maximal inhibition of lipogenesis by pST treatment in adipose tissue in vitro was observed on d 4 ( 68%) and d 7 (-69%). Similarly, fatty acid synthase activity declined during the treatment period, with the greatest change noted on d 7 (-26%). After withdrawal from treatment, lipogenesis gradually increased, returning to control values 7 d after withdrawal. Levels of IGF-I began to increase from d 1 to d 7 of treatment, continually decreased during withdrawal, and were normalized by the end of the withdrawal period. Plasma urea nitrogen concentrations decreased during treatment, increased during the withdrawal phase, and were normalized 4 d after the last pST treatment. Overall results indicate that most of the metabolic changes in response to pST occur within 1 wk of treatment and return to pretreatment values after 7 d of withdrawal from treatment. PMID- 10834578 TI - The effects of selection of pigs on growth rate vs leanness on histochemical characteristics of different muscles. AB - The microstructure of two type of muscles was studied in a selection experiment conducted with Dutch Large White pigs (boars and gilts) selected for either low backfat thickness (L-line) or fast growth (F-line). Second- and fourth-generation pigs were used to determine effects of selection on fiber type composition, fiber area, and capillary density in the longissimus lumborum (LL) and biceps femoris (BF) muscles. Immediately after slaughter samples were taken from the LL and BF muscles. The latter was divided into an inside (BFi) and outside (BFo) portion, which refer to the red and white portions of the biceps femoris. Serial sections were stained for ATPase (pH 4.60), succinate dehydrogenase (SDH), and alpha amylase-periodic acid shiff (PAS) to determine fiber type and capillary density. The LL and BFo muscles had predominantly type IIBw fibers, whereas the BFi muscle had a 2 to 4 times higher amount of type I fibers. In most muscles there were more type I and fewer type IIBw fibers in F- than in L-line pigs (P < .05), except in the LL muscle of second-generation pigs and in the red part of the BF muscle of fourth-generation pigs. In both selection lines lower type I and higher type IIBw percentages were found in muscles from gilts than in those from boars (P < .05). Capillary density and fiber area of L- and F-lines showed minor differences, which could be explained by differences in weight and age of the pigs of both lines. The results suggest that selection for low backfat thickness in pig breeding compared with increased growth rate resulted in fewer oxidative and more glycolytic muscle fibers. The magnitude of the effect depended on muscle type and duration of the selection period. PMID- 10834579 TI - Using live estimates and ultrasound measurements to predict beef carcass cutability. AB - Commercial slaughter steers (n = 329) and heifers (n = 335) were selected to vary in frame size, muscle score, and carcass fat thickness to study the effectiveness of live evaluation and ultrasound as predictors of carcass composition. Three trained personnel evaluated cattle for frame size, muscle score, fat thickness, longissimus muscle area, and USDA quality and yield grade. Live and carcass real time ultrasound measures for 12th-rib fat thickness and longissimus muscle area were taken on a subset of the cattle. At the time of slaughter, carcass ultrasound measures were taken at "chain speed." After USDA grade data were collected, one side of each carcass was fabricated into boneless primals/subprimals and trimmed to .64 cm of external fat. Simple correlation coefficients showed a moderately high positive relationship between 12th rib fat thickness and fat thickness measures obtained from live estimates (r = .70), live ultrasound (r = .81), and carcass ultrasound (r = .73). The association between estimates of longissimus muscle area and carcass longissimus muscle area were significant (P < .001) and were higher for live evaluation (r = .71) than for the ultrasonic measures (live ultrasound, r = .61; carcass ultrasound, r = .55). Three-variable regression equations, developed from the live ultrasound measures, explained 57% of the variation in percentage yield of boneless subprimals, followed by live estimates (R2 = .49) and carcass ultrasound (R2 = .31). Four variable equations using frame size, muscle score, and selected fat thickness and weight measures explained from 43% to 66% of the variation for the percentage yield of boneless subprimals trimmed to .64 cm. Live ultrasound and(or) live estimates are viable options for assessing carcass composition before slaughter. PMID- 10834580 TI - Carcass traits and retail display-life of chops from different goat breed types. AB - Four groups of goats, Boer x Spanish, straightbred Spanish, Spanish x Angora, and straightbred Angora were slaughtered at a constant age according to accepted industry procedures. At 24 h postmortem, various carcass yield and quality measurements were taken. At 48 h postmortem, one side from each carcass was fabricated into major wholesale cuts for dissection into percentage lean, bone, and fat. Rib chops from the opposite side were fabricated, packaged, and displayed in a retail case. Trained panelists evaluated the rib chops over 4 d of retail display for lean color, surface discoloration, and overall appearance; packages were opened and analyzed for off-odor on d 4. When slaughtered at constant ages, Angora goats had lighter (P < .05) live and hot carcass weights than all other breed types, and Boer x Spanish goats had heavier (P < .05) live and carcass weights than Spanish goats. Live and carcass weights for Boer x Spanish and Spanish x Angora goats did not differ (P > .05). Carcasses from Angora goats had considerably smaller (P < .05) longissimus muscle areas than all other breed types. Carcasses from Spanish goats had lower (P < .05) carcass conformation scores than carcasses from Boer x Spanish goats but did not differ (P > .05) from carcasses of the other two breed types. Carcasses from Angora goats had smaller (P < .05) leg circumferences than the carcasses from Boer x Spanish and Spanish x Angora breed types. In general, carcasses from Boer x Spanish and Spanish goats possessed higher (P < .05) percentages of lean and lower (P < .05) percentages of fat for the side than did carcasses from Spanish x Angora and Angora goats. When the Spanish x Angora carcasses were compared to the Angora carcasses, it seemed that the addition of the Spanish breeding tended to increase lean and decrease fatness for most side or primal comparisons. There were no (P > .05) breed type or breed type x day effects for lean color, surface discoloration, overall appearance or off-odor; however, day of display did influence (P < .05) these traits. Crossing Spanish with Angora goats may be an option to improve carcass characteristics over that of straightbred Angora. PMID- 10834581 TI - Riboflavin and niacin concentrations of bison cuts. AB - We analyzed the riboflavin and niacin contents of individual cuts from clod (triceps brachii), ribeye (longissimus thoracis), top round (semimembranosus), and top sirloin (gluteus medius) from 24 fed bison bulls. The bulls came from producers in the United States and Canada and had consumed concentrate diets plus hay free choice for at least 100 d. The mean riboflavin and niacin concentrations of all of the bison cuts combined were .094 and 1.910 mg/100 g wet weight, respectively. The riboflavin and niacin content values did not differ (P < .05) among the cuts of meat. Cuts from individual bulls were significantly different (P < .05) with regard to both riboflavin and niacin contents. Little variation was observed in riboflavin and niacin content of five bison from the same producer and two bison from another producer. These content values may be used in estimating the riboflavin and niacin content of bison meat. PMID- 10834582 TI - Effect of dietary lysine level and environmental temperature during the finishing phase on the intramuscular fat content of pork. AB - This study was designed to investigate the effects of dietary lysine level on the intramuscular fat content of the longissimus in finishing pigs reared at two environmental temperatures. Seventy-two hybrid gilts were individually penned and given ad libitum access to either a diet formulated to meet their lysine requirement (6.4 g/kg lysine) or a lysine-deficient diet (4.8 g/kg). Pigs were held at one of two environmental temperatures (thermoneutral [18 degrees C] or hot [32 degrees C]). The study was carried out between approximately 90 and 126 kg live weight; pigs in the thermoneutral and hot environments were on test for 5 and 7 wk, respectively. There were no interactions between dietary lysine level and environmental temperature. Dietary lysine content did not influence feed intake or average daily gain; however, pigs on the lysine-deficient diet had a poorer gain:feed ratio than those fed to requirement (P < .01). High environmental temperature decreased feed intake (P < .001) and average daily gain (P < .01) but improved gain:feed ratio (P < .01). Backfat at the 10th rib was increased and loin eye area and estimated percentage lean in the carcass were decreased for pigs on the lysine-deficient diet. The higher environmental temperature resulted in an increase in carcass length but had no effect on other carcass measurements or intramuscular fat. Feeding the lysine-deficient diet resulted in an increase of .55 percentage unit in longissimus intramuscular fat content (P < .01); however, there was no difference in subjective marbling scores between the diets. Warner-Bratzler shear force values were not affected by dietary lysine level or environmental temperature. Results from this study suggest that feeding of lysine-deficient diets at the end of the finishing period can increase intramuscular fat deposition under thermoneutral and hot conditions. PMID- 10834583 TI - Effect of ovarian phase and follicle quality on morphology and developmental capacity of the bovine cumulus-oocyte complex. AB - In order to study the effect of the follicular environment on the quality and developmental competence of cumulus-oocyte complexes (COC), COC were collected from nonatretic (NA), light atretic (LA), atretic (A), and heavily atretic (HA) follicles. Cumulus-oocyte complexes were also collected from early-luteal phase ovaries (EL), from late-luteal phase ovaries (LL), from follicular phase ovaries (F), and from ovaries from non-cyclic animals (NC). Each COC was assigned to one of three quality groups based on the appearance of the cumulus investment: 1) COC A: compact and bright, 2) COC-B: less compact and dark, and 3) COC-C: strongly expanded cumulus with dark spots. There was a high correlation between follicle quality and the distribution of the COC over the three COC qualities. The COC-A were mainly but not exclusively derived from NA follicles, COC-B were mainly derived from all classes of atretic follicles, and COC-C nearly exclusively originated from HA follicles. The developmental capacity of COC-A and COC-B, which was measured by in vitro embryo production, was consistent over the follicle qualities, except for COC-B obtained from HA follicles. They showed lower development (10.6%, P < .05) compared with COC-B from the other follicle qualities (18.9, 18.7, and 19.8%, respectively, for COC-B from NA, A, and LA follicles). The COC-B from atretic follicles produced more blastocysts (19.8%) than COC-A (12.7%, P < .05). The overall percentage of produced embryos per follicle class seemed to increase with increasing signs of atresia, except if the COC were derived from HA follicles. This increased percentage of embryos was, however, not due to a better quality of COC, but to a higher percentage of COC-B coming from these follicles. The stage of the cycle had no effect on the distribution of the COC over the three COC qualities or on the developmental capacity of COC-A or COC-B, except for COC-A from EL ovaries, which produced more (P < .05) blastocysts than COC-A from the other luteal phases (12.5% vs approximately 8%). A follow-up study was performed trying to elucidate why COC-B possess a higher developmental potency than COC-A. The answer was sought in the oocyte maturation. At several time points during maturation, oocytes were evaluated for their nuclear stage. At all time points COC-B seemed to be a few hours ahead of COC-A, and after 24 h of maturation more COC-B had reached the metaphase-2 stage. This might mean that COC-A need a longer maturation period than COC-B. PMID- 10834584 TI - Beta-carotene uptake and changes in ovarian steroids and uterine proteins during the estrous cycle in the canine. AB - The uptake of beta-carotene by reproductive tissues and the effects of beta carotene on reproductive function in the dog are unknown. We studied the uptake of beta-carotene by blood, corpus luteum, and uterine endometrium and the role of dietary beta-carotene in influencing ovarian steroid and uterine protein production during the estrous cycle in the dog. Mature female Beagle dogs (n = 56) were fed diets containing 0, 2, 20, or 50 mg of beta-carotene daily for approximately 6 wk before estrus detection. Blood was sampled at regular intervals from estrus through d 45 after ovulation (d 0 = ovulation), when laparotomy was performed. The ovaries were obtained for the isolation of corpus luteum. The uterus was flushed with phosphate-buffered saline and the endometrium obtained by scraping. Beta-carotene was not detectable in plasma, corpus luteum, or endometrium of unsupplemented dogs. However, beta-carotene and alpha-carotene in plasma, corpus luteum, and uterine endometrium increased in a dose-dependent manner. Alpha-carotene made up a high percentage of total carotenoids even though the alpha-carotene content in the dietary source was very low. Dogs fed 50 mg of beta-carotene had significantly higher concentrations of plasma progesterone between d 12 and 26 compared with unsupplemented dogs. Dietary beta-carotene did not influence plasma estradiol-17beta and total uterine proteins. Therefore, beta carotene is absorbed into plasma, corpus luteum, and uterine endometrium of dogs. Furthermore, dietary beta-carotene increased plasma progesterone concentrations during the estrous cycle. It is possible that dietary beta-carotene may improve reproductive function in the canine. PMID- 10834585 TI - Ovarian and endocrine characteristics during an estrous cycle in Angus, Brahman, and Senepol cows in a subtropical environment. AB - To determine breed differences in ovarian function and endocrine secretion, daily rectal ultrasonography was conducted on multiparous lactating Angus (temperate Bos taurus; n = 12), Brahman (tropical Bos indicus; n = 12), and Senepol (tropical Bos taurus; n = 12) cows during an estrous cycle in summer. Blood was collected daily to quantify plasma concentrations of FSH, LH, progesterone, estradiol, GH, insulin-like growth factor (IGF)-I, IGF-II, IGF binding proteins (IGFBP), insulin, glucose, and plasma urea nitrogen (PUN). Numbers of small (2 to 5 mm), medium (6 to 8 mm), and large follicles (> or = 9 mm) were greater (P < .05) in Brahman than in Angus and(or) Senepol cows. Length of the estrous cycle (SEM = .6 d) was similar (P > .10) among Senepol (20.4 d), Angus (19.5 d), and Brahman (19.7 d) cows. Senepol cows had greater (P < .05) diameters of the corpus luteum (CL) and a delayed regression of the CL as compared with Angus cows. The secondary surge of FSH (between d 1 and 2; d 0 = estrus) was greater in Angus than Brahman or Senepol cows (breed x day, P < .05). Between d 2 and 14 of the estrous cycle, concentrations of progesterone, LH, IGF-II, and binding activities of IGFBP-3, IGFBP-2, and the 27- to 29-kDa IGFBP in plasma did not differ (P > .10) among breeds. Concentrations of GH, IGF-I, insulin, and PUN were greater (P < .001) and binding activities of the 22-kDa and 20-kDa IGFBP tended (P < .10) to be greater in plasma of Brahman than in Angus or Senepol cows. Plasma glucose concentrations were greater (P < .05) in Senepol than in Brahman or Angus cows. In conclusion, Brahman (Bos indicus) and Senepol cows (tropical Bos taurus) had greater numbers of follicles in all size categories and greater diameter of CL than Angus (temperate Bos taurus) cows. These ovarian differences may be due to changes in the pattern of secretion of FSH, insulin, IGF-I, and GH but not LH, IGF-II, or IGFBP-2 or -3. PMID- 10834586 TI - Inhibition of prolactin in the last trimester of gestation decreases mammary gland development in gilts. AB - Prolactin is required for mammary development in various species but its possible role for mammogenesis in pigs is not known. The goal of the present study was therefore to determine the effect of prolactin inhibition by bromocriptine during the last third of gestation on mammary gland development in gilts. Twenty-eight primigravid gilts were assigned as controls (n = 15) or received 10 mg of bromocriptine orally thrice daily (n = 13) from d 70 to 110 of gestation. Jugular blood samples were collected on d 70 of gestation and every 8 d thereafter and were assayed for prolactin, IGF-I, estradiol, and progesterone. Gilts were slaughtered on d 110 of gestation and fetuses were counted and weighed. One row of mammary glands was used for dissection of parenchymal and extraparenchymal tissues and for determination of DNA, RNA, dry matter, protein, and fat contents. Tissue from the other row was used for measures of prolactin receptor number and affinity. Concentrations of prolactin were drastically reduced throughout the bromocriptine treatment period (P < .001), whereas there was no overall treatment effect on progesterone and IGF-I levels (P > .10). Total weight and extraparenchymal tissue weight of the mammary glands were unaffected by treatment (P > or = .1), but weight of parenchymal tissue, total DNA, and total RNA decreased (P < .01) with bromocriptine treatment. Percentages of fat and dry matter in parenchymal tissue increased with bromocriptine treatment (P < .01) and the percentage of protein decreased (P < .01). Number of prolactin receptors in parenchymal tissue decreased with bromocriptine treatment (P < .001) and receptor affinity increased (P < .001). Average fetal weight was lower in gilts receiving bromocriptine than in control gilts (P = .05), but fetal number did not differ (P > .1). These results clearly demonstrate that prolactin is essential for normal mammary gland development and can affect fetal growth during the last third of gestation in gilts. PMID- 10834587 TI - Technical note: recombinant LHRH fusion protein suppresses estrus in heifers. AB - A recombinant luteinizing hormone-releasing hormone (LHRH) fusion protein was evaluated for its effectiveness in suppression of estrus in heifers. Eight heifers were randomly assigned to two equal treatment groups. Treatments consisted of recombinant ovalbumin-LHRH-7 or recombinant ovalbumin (control). This recombinant chimeric fusion protein consisted of ovalbumin with seven LHRH peptides (ovalbumin-LHRH-7). The plasmid for this protein was expressed in E. coli and was collected and purified as an insoluble protein. One milligram of the respective proteins was suspended in 2 mL of Z-Max adjuvant and administered by intramammary injection three times at 7-wk intervals. Luteinizing hormone releasing hormone antibody binding was elevated in heifers treated with ovalbumin LHRH-7 compared to ovalbumin-treated heifers (P < .05). Serum progesterone concentrations (< 1 ng/mL) indicate that the estrous cycle of the four heifers treated with ovalbumin-LHRH-7 was suppressed for a time period ranging from 60 to 238 d, which was different from control heifers (P < .01). Serum progesterone for the control heifers continued to exhibit cyclic profiles over the experimental period. This preliminary study in heifers demonstrated that a chimeric LHRH fusion protein induced elevated concentrations of circulating LHRH antibodies that suppressed estrus for an average of 122 +/- 41 d. PMID- 10834588 TI - Growth of nursing pigs related to the characteristics of nursed mammary glands. AB - The purpose of this study was to determine growth performance of nursing pigs in relationship to teat order and to observe teat preference by pigs. In the first experiment, litter size of 13 primiparous sows was adjusted to 9 (8.7 +/- 1.5) pigs and teat order of each litter was observed on the day before slaughter. Another group of eight sows was killed on d 0 (within 12 h after farrowing). In the second experiment, litter size was adjusted to 9 (8.9 +/- 1.4) pigs for 20 primiparous sows and teat order for each litter was observed 1 d before slaughter. The weights of sows and individual pigs were recorded at farrowing, weekly, and on the day before slaughter. Mammary glands were collected at slaughter on d 21 of lactation and trimmed of skin and the extraneous fat pad. Individual glands were separated, weighed, and ground for measurement of dry matter, dry fat-free tissue, protein, fat, ash, and DNA contents. Middle mammary glands had the greatest wet weight among glands obtained within 12 h after weaning (P < .05). For sows completing the 21 d lactation, only glands known to have been nursed were included in the data sets. Greater than 60% of the first four pairs of mammary glands were nursed, and less than 40% of the seventh and eighth glands were nursed by pigs during lactation. Pigs that nursed the first five pairs of anterior glands gained faster than pigs nursing the remaining glands. The first five pairs of anterior glands had greater wet and dry weights, and greater protein and DNA contents compared with the remaining glands. Pigs that nursed heavier glands gained weight faster (r = .68, P = .0001), and those heavier glands contained greater amounts of protein (r = .98, P = .0001) and DNA (r = .66, P = .0001). Variation in weight gain of pigs nursing the anterior and middle glands was not statistically significant. The functional superiority of anterior and middle glands was positively correlated with body weight gain of nursing pigs. PMID- 10834589 TI - The effects of mutated skeletal ryanodine receptors on hypothalamic-pituitary adrenal axis function in boars. AB - The objectives of the current experiment were to determine whether boars heterozygous for the mutation in skeletal ryanodine receptors (sRyR), known to cause porcine stress syndrome, differed from wild-type boars in hypothalamic pituitary-adrenal axis (HPA) function. We have examined basal plasma ACTH, cortisol, and corticosteroid-binding globulin (CBG) concentrations; plasma ACTH and cortisol responses to a nose-snare stressor and at slaughter; dexamethasone suppression of plasma ACTH and cortisol concentrations; and glucocorticoid receptor (GR) density in the pituitary gland, hippocampus, hypothalamus, and frontal cortex. We have also examined carcass yields, composition, and meat quality to determine whether differences in HPA activity were accompanied by an increased incidence of meat quality characteristics associated with pale, soft, exudative (PSE) meat. Thirty boars either heterozygous or wild-type (n = 15 per genotype) for mutated sRyR were tested for HPA function at 7 mo of age. Heterozygous boars had lower basal plasma ACTH (P < .05) and cortisol (P < .04) concentrations. Integrated basal plasma ACTH and cortisol levels were also lower (P < .05 and P < .005, respectively). Genotype had no significant effect on basal CBG, stressor-induced (nose snare or slaughter) or dexamethasone suppression of plasma ACTH or cortisol concentrations. No differences in immunoreactive GR levels were found in the pituitary gland or any brain region examined. We did find a significant, negative correlation (r = -.62, P < .02) between peak (0800) basal plasma ACTH concentrations and hippocampal GR levels. The alterations in basal HPA function in heterozygous boars were accompanied by lighter body weights (P < .03), decreased carcass fat depth (P < .04), and increased carcass lean yields (P < .02). There was a higher incidence of meat quality characteristics associated with PSE meat in heterozygous boars indicated by higher carcass temperatures (P < .04) and meat brightness (P < .0001) with lower carcass pH at slaughter (P < .03) and after chilling (P < .003). In conclusion, we have found differences in basal and not stressor-induced HPA function between boars heterozygous and wild-type for mutated sRyR. This altered basal HPA activity was accompanied by an increased incidence of meat quality aspects associated with PSE meat in heterozygous boars. PMID- 10834590 TI - Effects of pasture applied biosolids on performance and mineral status of grazing beef heifers. AB - Angus x Hereford heifers (n = 50) were randomly assigned to bahiagrass pastures treated with biosolids varying in mineral content and evaluated for mineral status, with special attention to Cu. Biosolids and NH4NO3 were all applied at the rate of either 179 kg N/ha (X) or twice this (2X). Fertilizer was applied to .81-ha pastures for the following treatments: 1) Baltimore biosolids (1X = 179 kg N/ha); 2) Baltimore biosolids (2X = 358 kg N/ha); 3) Tampa biosolids (1X = 179 kg N/ha); 4) Tampa biosolids (2X = 358 kg N/ha); or 5) control NH4NO3 (1X = 179 kg N/ha) applied at two times. Pastures were divided into five blocks with each treatment represented once in each block. Copper loads varied from 8.8 to 42.2 kg/ha, and Mo loads varied from .27 to 1.11 kg/ha. Heifers (two per pasture) grazed their assigned pastures exclusively for 176 d. Liver biopsies were taken from all animals at d 1, 99, and 176, and blood samples on d 1, 50, 99, 135, and 176. Liver and plasma were analyzed for selected mineral contents, and blood was analyzed for hemoglobin and hematocrit. Experimental animals were generally low in mineral status when assigned to pastures and deficient in Se and P. By d 50, plasma Ca, Mg, Se, P, and Zn were adequate for all treatments. Plasma Cu declined (P < .03) for all treatments from d 50 to 176. Plasma Cu reflected depleted liver Cu storage, with the two Tampa and highest Baltimore treatment means lower in plasma Cu than the control at 176 d. Liver Fe concentrations were adequate for all treatments, and Mo concentrations (< 2.18 mg/kg) did not approach levels indicative of toxicity. Liver Cu declined (P < .05) with time for all treatments. By d 99, animals receiving the two Baltimore treatments and the lowest Tampa application rate had lower (P < .05) liver Cu than the control, and all treatments were lower at 176 d. The decline of animal Cu status (liver and plasma) reflects the low Cu status of bahiagrass and the possibility of high forage S (.30 to .47%) interfering with Cu metabolism. Forage Mo was low but was slightly higher in biosolids-treated pastures. High levels of biosolids applications to bahiagrass pastures were not detrimental to mineral status except Cu, which had a tendency to decline in plasma and for all biosolids treatments declined in liver. PMID- 10834591 TI - The effect of physical form of orchardgrass hay on the passage of particulate matter through the rumen of sheep. AB - Four Texel wethers (60 to 64 kg) fitted with ruminal and duodenal cannulas were used to study the kinetics of particulate matter in the rumen and the series of processes involved in their selection and passage. They were fed, in eight equal meals, 1,200 g of a mixture of a chopped orchardgrass hay and ground (8-mm screen) and pelleted orchardgrass hay in 90/10, 50/50, 30/70, or 10/90 ratios, according to a 4 x 4 Latin square design. The particle size distributions in feed, chewed feed, and ruminal, reticular, and duodenal digesta were determined by a wet-sieving procedure. Indigestible lignin was used as an internal marker to trace the passage of particles through the rumen. Digesta flow measurement was performed using the double-marker technique. We used a three-pool model, which partitions particles through the large, medium, and small particle pools, to determine the passage of lignin through those pools. Particle pool sizes and rumen and pool mean retention times (MRT) of lignin and of the rumen MRT of an ideal marker introduced separately in each pool were corrected for the "filter bed" effect. Grinding and pelleting of hay decreased the MRT of the indigestible lignin pool in the rumen. Particle MRT decreased and then reached a plateau with increased proportion of ground/pelleted hay in the diet. The diet with a ratio of 50/50 of chopped and ground/pelleted hay was the most favorable for the exit of particles from the rumen because of both a higher outflow rate from the rumen of particles eligible to exit and a sufficient comminution rate of larger particles to supply particles that were able to pass. For all diets, the large-particle comminution rate was always higher than the small-particle outflow rate, indicating that comminution was not the limiting step for passage. These results were the consequence of the curvilinear and opposite evolutions of both the particulate lignin pool in, and outflow from, the rumen. Those results contribute to an improved explanation of the mechanisms involved in the outflow of particles from the rumen. PMID- 10834592 TI - Splanchnic nitrogen metabolism by growing beef steers fed diets containing sorghum grain flaked at different densities. AB - The objective of this study was to determine effects of processing method, dry rolled (DR) vs steam-flaked (SF), and degree of processing (flake density, FD) of SF sorghum grain on splanchnic (gut and liver) N metabolism by growing steers. Diets contained 77% sorghum grain either DR or SF at densities of 437, 360, and 283 g/L (SF34, SF28, and SF22, respectively). Eight crossbred steers (340 kg initial BW), implanted with indwelling catheters into portal, hepatic, and mesenteric veins and the mesenteric artery, were used in a randomized complete block design. Blood flows and net output or uptake of ammonia N, urea N (UN), and alpha-amino N (AAN) were measured across portal-drained viscera, hepatic, and splanchnic tissues. Plasma arterial, portal, and hepatic concentrations of individual amino acids were also measured. Decreasing FD linearly increased (P = .04) net absorption of AAN (51, 73, and 78 g/d for SF34, SF28, and SF22, respectively) and transfer (cycling) of blood UN to the gut (49, 48 and 64 g/d; P = .02). Net UN cycling averaged 38% of N intake across all diets. Hepatic uptake of AAN or UN synthesis, and splanchnic output of AAN and UN, were not altered by FD. Lowering FD linearly increased (P < or = .02) portal-arterial concentration differences for blood AAN and UN and plasma arterial concentrations for alanine. Steers fed SF compared to DR tended to have greater (P = .11) blood UN cycling (percentage of hepatic synthesis; 64 vs 50%) and decreased (P = .03) net splanchnic UN output (30 vs 50 g/d), but other net fluxes of N were not altered across splanchnic tissues. Steam-flaking compared to dry-rolling tended to decrease (P = .12) portal, but not hepatic, blood flow and increased (P < .01) hepatic-arterial concentration differences for blood UN. Except for a decrease (P = .01) in hepatic-arterial concentration differences of glutamine, plasma amino acid concentrations were not altered by feeding SF vs DR sorghum. Processing method (steam-flaking vs dry-rolling) or increasing the degree of processing (by decreasing FD) of SF sorghum grain resulted in greater transfer of blood UN to the gut. Reducing FD also linearly increased the absorption of AAN by growing steers, which explains (in part) published responses of superior performance by steers fed SF grains. PMID- 10834593 TI - Net absorption and hepatic metabolism of glucose, L-lactate, and volatile fatty acids by steers fed diets containing sorghum grain processed as dry-rolled or steam-flaked at different densities. AB - We determined the effect of processing method (dry-rolled [DR] vs steam-flaked [SF]) and degree of processing (flake density; FD) of SF sorghum grain on splanchnic (gut and liver) metabolism of energy-yielding nutrients by growing steers. Diets contained 77% sorghum grain, either DR or SF, with SF at densities of 437, 360, or 283 g/L (SF34, SF28, or SF22). Eight multicatheterized steers (340 kg initial BW) were used in a randomized complete block design. Net output or uptake of glucose, L-lactate, VFA, and beta-hydroxybutyrate (BHBA) were measured across portal-drained viscera (PDV), liver, and splanchnic (PDV plus liver) tissues. Net absorption of glucose across PDV was negative and similar for all treatments (average of -104 g/d). Decreasing FD of SF sorghum grain linearly increased (P < or = .04) net absorption and splanchnic output of L-lactate by 20 and 130%, respectively, and hepatic synthesis (P = .06) and splanchnic output (P = .01) of glucose by 50%. Reducing FD did not alter output or uptake of acetate or n-butyrate by gut and liver tissues, but linearly decreased (P = .06) splanchnic output of BHBA by 40%. Net absorption (P = .18) and splanchnic output (P = .15) of propionate tended to be increased linearly by 50% with decreasing FD. Neither processing method (SF vs DR) nor degree of processing (varying FD) altered hepatic nutrient extraction ratios or estimated net absorption and splanchnic output of energy. Maximal contribution of propionate, L-lactate, and amino acids (alpha-amino N) to gluconeogenesis averaged 49, 11, and 20%, respectively. Feeding steers SF compared to DR diets did not alter net output or uptake of energy-yielding nutrients across splanchnic tissues, except net absorption of acetate tended to be greater (P = .13) for steers fed DR. Increasing degree of grain processing in the present study, by incrementally decreasing FD, tended to linearly increase the net absorption of glucose precursors (propionate and lactate), resulting in linear increases in synthesis and output of glucose by the liver to extrasplanchnic tissues (e.g., muscle). PMID- 10834594 TI - Net portal appearance of volatile fatty acids in sheep intraruminally infused with mixtures of acetate, propionate, isobutyrate, butyrate, and valerate. AB - The net portal appearance of volatile fatty acids (VFA) was investigated in four ruminally fistulated and multicatheterized sheep. During the experiments, the sheep were fed once every hour for 14 h and intraruminally infused with mixtures of VFA for the 12 h commencing 2 h after the initiation of the hourly feeding protocol. Paired arterial and portal blood samples were obtained hourly during the last 6 h of the experiments. In the control treatment (1), only water was infused intraruminally. In Treatments 2 through 4, the intraruminal infusion rates of propionate (40 mmol/h), isobutyrate (5 mmol/h), and valerate (5 mmol/h) were unchanged. In Treatments 2, 3, and 4, the acetate infusion rate was 100, 60, and 20 mmol/h, respectively, and the butyrate infusion rate was 10, 30, and 50 mmol/h, respectively. Thus, the infusion rate of VFA carbon was constant across Treatments 2 through 4. Portal recovery estimated from the increased net portal appearance in Treatments 2 through 4 compared to the control treatment was 85% for propionate and 60% for isobutyrate, and these recoveries were unaffected by treatment. The portal recovery of butyrate increased (from 21 to 32%) with increasing infusion rate of butyrate and decreasing infusion rate of acetate, as did the portal recovery of valerate (from 14 to 31%). The portal recovery of acetate was 55%, when measured as net portal appearance. Thus, it seems that the capacity for beta-oxidation in ruminal epithelium is limited, which would explain the increasing portal recovery of butyrate and valerate with increasing infusion rate of butyrate, when infusion rate of VFA carbon is unchanged. PMID- 10834595 TI - Net flux of nonesterified fatty acids, cholesterol, triacylglycerol, and glycerol across the portal-drained viscera and liver of pregnant ewes. AB - The objective of this study was to determine the pattern of nutrient flux across portal-drained viscera (PDV) and liver in ewes with varying numbers of fetuses. Catheters were placed in the hepatic portal vein, a branch of the hepatic vein, a mesenteric vein, and the abdominal aorta of each ewe. Plasma flow and net cholesterol, nonesterified fatty acids (NEFA), and glycerol release across the PDV and liver were determined prior to exposure to rams. Ewes were subsequently mated. Two ewes were not pregnant, six ewes gave birth to singles, and 11 ewes gave birth to twins. Additional measurements were taken 103, 82, 61, 39, 19, and 6 d before parturition. There was a net PDV uptake of nonesterified cholesterol in the nonpregnant ewes and a net release in the ewes with singles and twins. Net nonesterified cholesterol hepatic release did not differ with days from parturition (P = .77). There was a net hepatic release of nonesterified cholesterol in the ewes with twins and a net hepatic uptake in the ewes with singles and in nonpregnant ewes (P = .03). There was a net PDV release of NEFA; however, it did not differ with litter size (P = .59) or days from parturition (P = .63). Hepatic NEFA uptake increased with litter size (P = .03) and increased as gestation progressed (P = .006). There was an interaction (P = .04) between litter size and days from parturition for net PDV glycerol release. Net PDV glycerol release in the nonpregnant ewes decreased over time, but release in pregnant ewes tended to increase over time. Hepatic glycerol uptake increased with litter size and increased as gestation progressed. There was a net PDV uptake of triacylglycerol, but it did not differ with litter size (P = .11) or with days from parturition (P = .06). There was a net hepatic release of triacylglycerol, but it did not differ with litter size (P = .59) or with days from parturition (P = .67). Liver utilization of glycerol and NEFA as substrates for metabolism increases as pregnancy progresses. In the nonpregnant ewe, the combined contribution of glycerol and NEFA carbon accounted for 10% of the carbon taken up by the liver, and in ewes pregnant with twins, the combined contribution accounted for 42% of the carbon uptake 19 d before parturition. In conclusion, these data demonstrate NEFA are an important metabolite when determining carbon balance across the liver and their relative contribution to carbon balance increases as pregnancy progresses. PMID- 10834596 TI - Rapid communication: nucleotide sequence of the river buffalo kappa-casein cDNA. PMID- 10834597 TI - Rapid communication: nucleotide sequence of the river buffalo beta-casein cDNA. PMID- 10834598 TI - Rapid communication: physical and genetic mapping of the Peroxisome Proliferator Activated Receptor gamma (PPAR-gamma) gene to porcine chromosome 13. PMID- 10834599 TI - Rapid communication: the ovine cDNA encoding interferon-stimulated gene product 17 (ISG17). PMID- 10834600 TI - Rapid communication: linkage mapping of the porcine Agouti gene. PMID- 10834601 TI - Rapid communication: nucleotide sequences of the bovine, caprine, and ovine beta3 adrenergic receptor genes. PMID- 10834602 TI - Antimicrobial prophylaxis in dermatology. AB - Few circumstances in dermatology warrant antimicrobial prophylaxis. In cutaneous surgery postoperative infections are too infrequent and insufficiently severe to justify preventive antibiotics, except rarely. Petrolatum is as effective as, and cheaper than, topical antibiotic ointment to cover surgical wounds. In patients with numerous staphylococcal skin infections, oral clindamycin 150 mg every day for 3 months safely reduces further episodes. For recurrent cellulitis, oral penicillin or erythromycin 250 mg twice daily or monthly intramuscular benzathine penicillin decreases subsequent attacks. In patients with frequent episodes of genital or labial herpes simplex an antiviral agent such as valacyclovir 500 mg to 1 g every day is effective as a suppressant. PMID- 10834603 TI - Lyme borreliosis. AB - Lyme borreliosis is a worldwide, multistage, and multi-system disease caused by borrelia spirochetes, which are transmitted by ixodes ticks. It is focally endemic in temperature climates of the northern hemisphere. Primary erythema migrans occurs at the site of inoculation. Secondary erythema migrans occurs at sites of hematogenous dissemination. Variations in genospecies account for variations in presentation, including borrelial lymphocytoma. Disseminated disease includes constitutional signs and symptoms, intermittent oligoarticular arthritis, meningitis, cranial neuritis, radiculoneuropathy, encephalopathy, atrioventricular block, and myopericarditis. Late persistent disease includes acrodermatitis chronica atrophicans, chronic arthritis, neurological impairment, and fatigue. There can be difficulties with both clinical and laboratory diagnosis. First-line oral therapies for early uncomplicated disease are doxycycline and amoxicillin. First-line intravenous therapy for complicated or resistant disease is ceftriaxone. Prevention includes avoiding tick habitats, dressing sensibly, judicious use of repellants, and early removal of imbedded ticks. Vaccination is indicated only for frequent or prolonged exposure to tick infested habitat. PMID- 10834604 TI - The immune compromised host in the twenty-first century: management of mucocutaneous infections. AB - Infectious diseases encountered in dermatology have changed tremendously during the past few decades with the emergence of the immunocompromised host. This change is a result of the human immunodeficiency virus epidemic, use of immunomodulating drugs, bone marrow transplantation, increasing prevalence of diabetes mellitus, and an aging population. New pathogens have been discovered and new disorders have occurred. In the compromised host, infection can be more aggressive and widespread locally, be caused by opportunistic pathogens, and be disseminated hematogenously from or to the skin. The prevalence of nonmelanoma skin cancer has increased, and squamous cell carcinomas can be more aggressive with more rapid local growth as well as frequency of metastasis. PMID- 10834605 TI - Common and unusual cases seen by an inpatient dermatology consult service. AB - This article describes common consult requests and presents case studies from the dermatology consult service of an academic hospital. PMID- 10834606 TI - The application of flow cytometry to histocompatibility testing. AB - Flow cytometry is a powerful technique that enables the sensitive and quantitative detection of both cellular antigens and bound biological moieties. This article reviews how flow cytometry is increasingly being used as histocompatibility laboratories for the analysis of antibody specificity and HLA antigen expression. A basic description of flow cytometry principles and standardisation is given, together with an outline of clinical application in the areas of pre-transplant cross-matching, antibody screening, post-transplant antibody monitoring and HLA-B27 detection. It is concluded that flow cytometry is a useful multi-parametric analytical tool, yielding clinical benefit especially in the identification of patients at risk of early transplant rejection. PMID- 10834607 TI - Chemokine and chemokine receptor expression by liver-derived dendritic cells: MIP 1alpha production is induced by bacterial lipopolysaccharide and interaction with allogeneic T cells. AB - Dendritic cells (DC) are highly-specialized antigen-presenting cells (APC), that initiate and modulate immune responses. Their specialized migratory and tissue homing properties are regulated by small molecular weight proteins (chemokines) that govern leukocyte migration and activation. Little is known about the capacity of liver DC to produce or respond to chemokines. Here we examined chemokine and chemokine receptor (CR) gene expression in both immature DC progenitors (DCp) and comparatively mature DC generated from mouse liver. Factors affecting production of the chemokine macrophage inflammatory protein (MIP) 1alpha, and the influence of MIP-1alpha on liver DC migration were also investigated. Dendritic cells were propagated in response to granulocyte macrophage colony stimulating factor (GM-CSF) +/- interleukin (IL)-4 from bone marrow (BM) cells or liver non-parenchymal cells (NPC) isolated from normal mice, or from mice treated with the hematopoietic growth factor Flt3 ligand (FL). Their phenotype and allostimulatory function were assessed by monoclonal antibody (mAb) staining and flow cytometry, and by the capacity to induce mixed leukocyte reactions, respectively. Specific chemokine and CR gene expression was studied using the RNase protection assay (RPA). Production of MIP-1alpha was determined by enzyme-linked immunoabsorbent assay (ELISA), and the migratory activity of liver DC induced by MIP-1alpha quantitated using microchemotaxis chambers. Like DC generated simultaneously from BM, liver-derived DC expressed mRNA for a variety of CC and CXC chemokines. RANTES (regulated upon activation, normal T cell expressed and secreted) transcripts were the most strongly expressed. Gene transcripts for the receptor CCR1, that binds RANTES and MIP-1alpha were also readily detected, as was CCR2, the receptor for the monocyte chemotactic proteins (MCP)1-4. No major differences in chemokine or CR mRNA expression were detected between immature and more mature liver DC. MIP-1alpha production by liver-derived DC was stimulated by bacterial lipopolysaccharide (LPS), and high levels were also detected in co-cultures of hepatic DC and allogeneic T cells. Chemotactic migration of liver-derived DC was stimulated by MIP-1alpha. Thus, liver-derived DC express mRNA for several CC and CXC chemokines and their receptors that may play key roles in the regulation of hepatic inflammatory responses. Production of MIP-1alpha by liver DC, and their migratory responses to this chemokine, suggest that MIP-1alpha and other chemokines may play significant roles in the regulation of liver DC function and in interactions of liver DC with other leukocytes, under normal and inflammatory conditions. PMID- 10834608 TI - Kinetic analysis of microchimerism induced by intrathymic injection of allogeneic splenocytes in mice. AB - Allograft survival facilitated by intrathymic (i.t.) injection of allogeneic cells have shown that modifications of T-cell development induce specific tolerance. One hypothesis is that the resulting microchimerism may play a role in preparing the host immune system for the allograft. To investigate whether the deliberate introduction of allogeneic splenocytes into the thymus of adult mice allows the establishment of a lasting donor/recipient microchimerism, a full allogeneic mouse system (H-2 and Mls) with additional sex mismatch was used. Male cells injected into female mice were detected using an optimized nested polymerase chain reaction which specifically amplifies the SRY gene with a sensitivity of 1/10(4). After i.t. injection, donor cells were observed early both in the lymph nodes and spleen (75 and 25% of mice, respectively). They were still present on day 6, although preferentially in the thymus (100% of mice) than in the lymph nodes (50% of mice) or in the spleen (22% of mice). After intraperitoneal (i.p.) or subcutaneous (s.c.) injection, donor cells were early (2 h) but transiently detected in the thymus, since on day 6 they were detected in 0 and 17% of mice after i.p. and s.c. injection, respectively. Kinetics of donor-cell detection was similar both in the spleen and lymph nodes with a clear decrease in the percentage of mice with donor-cell detection between day 2 and day 6 (20 and 17% of positive mice for the spleen after i.p. and s.c. injections, respectively--20 and 33% of positive mice for the lymph nodes after i.p. and s.c. injections, respectively). Our results clearly show that i.t. injection of allogeneic splenocytes induces a microchimerism which is both more lasting and detected in a higher percentage of mice than by the i.p. and s.c. routes, both at the central (thymus) and peripheral (spleen) levels. PMID- 10834609 TI - T cell deletion and unresponsiveness induced by intrathymic injection of staphylococcal enterotoxin B. AB - Intrathymic injection of alloantigens appears to be the most efficient route to induce alterations of T cell reactivity. In the present study, we explored the modifications of Vbeta8.1, 8.2 T cell population and T cell reactivity in the thymus and in the spleen induced by intrathymic injection of staphylococcal enterotoxin B to adult mice. Vbeta8 antigen expression was investigated by flow cytometry analysis. T Cell reactivity was studied in vitro by the proliferative response to SEB. SEB induced a significant reduction in the percentage of mature Vbeta8+ T cells in the thymus (days 7-14), and in the spleen (days 7-28). Interestingly, this depletion occurs in the CD4- CD8+ cells in the thymus whereas in the CD4+ CD8- cells in the spleen. In parallel, the proliferative response to SEB but not to SEA was significantly decreased in the thymus on days 7 and 14, and in the spleen from day 7 to day 28. Moreover, this unresponsiveness was more pronounced in the spleen than in the thymus. Anergy was SEB-specific and fully reversed by exogenous IL-2. SEB injected intrathymically induced significantly more pronounced and more durable T cell alterations than intraperitoneal and subcutaneous injections. This may be related to the observation that after i.t. injection, SEB was detected both at a higher amount and for a longer period in the central and peripheral compartments. Our results clearly demonstrate that the intrathymic route is definitely the most efficient to induce not only thymic but also peripheral pivotal immune alterations in our model. PMID- 10834610 TI - Comparative analysis of dendritic cell density and total number in commonly transplanted organs: morphometric estimation in normal mice. AB - Dendritic cells (DC) are considered to be the major cell type responsible for induction of primary immune responses. While they have been shown to play a critical role in eliciting allosensitization via the direct pathway, there is evidence that maturational and/or activational heterogeneity between DC in different donor organs may be crucial to allograft outcome. Despite such an important perceived role for DC, no accurate estimates of their number in commonly transplanted organs have been reported. Therefore, leukocytes and DC were visualized and enumerated in cryostat sections of normal mouse (C57BL/10, B10.BR, C3H) liver, heart, kidney and pancreas by immunohistochemistry (CD45 and MHC class II staining, respectively). Total immunopositive cell number and MHC class II+ cell density (C57BL/10 mice only) were estimated using established morphometric techniques--the fractionator and disector principles, respectively. Liver contained considerably more leukocytes (approximately 5-20 x 10(6)) and DC (approximately 1-3 x 10(6)) than the other organs examined (pancreas: approximately 0.6 x 10(6) and approximately 0.35 x 10(6); heart: approximately 0.8 x 10(6) and approximately 0.4 x 10(6); kidney approximately 1.2 x 10(6) and 0.65 x 10(6), respectively). In liver, DC comprised a lower proportion of all leukocytes (approximately 15-25%) than in the other parenchymal organs examined (approximately 40-60%). Comparatively, DC density in C57BL/10 mice was heart > kidney > pancreas >> liver (approximately 6.6 x 10(6), 5 x 10(6), 4.5 x 10(6) and 1.1 x 10(6) cells/cm3, respectively). When compared to previously published data on allograft survival, the results indicate that the absolute number of MHC class II+ DC present in a donor organ is a poor predictor of graft outcome. Survival of solid organ allografts is more closely related to the density of the donor DC network within the graft. PMID- 10834611 TI - Donor-derived soluble HLA plasma levels can not be used to monitor graft rejection in heart transplant recipients. AB - OBJECTIVE: Increased levels of both donor- and recipient-derived HLA class I molecules (sHLA-I) can be found in serum or plasma of transplanted patients during rejection. Earlier data indicate that levels of donor-derived sHLA-I (dsHLA-I) correlate better with graft rejection than total sHLA Class I (Zavazava N, Kraatz E, Gassel AM, Muller-Ruchholtz W. Plasma MHC class I expression in cardiac graft patients: donor-specific soluble antigen in a pre-sensitized graft patient. Transplant Proc 1991;23:2258-2260; Claas FHJ, Jankowska-Gan E, DeVito LD, et al. Monitoring of heart transplant rejection using a donor-specific soluble HLA class I ELISA. Hum Immunol 1993;37:121). Therefore, quantifying donor derived soluble counterparts of HLA Class I (sHLA-I) in the plasma of the recipient may offer a new possibility for non-invasive monitoring of rejection after organ transplantation. METHODS: In an extended study with 34 heart transplant recipients, we used sHLA-I specific ELISAs to monitor donor-derived soluble sHLA-A2, -A3, -A9, -B7, -B12 and B51. RESULTS: The assays were sensitive enough to detect dsHLA Class I in plasma of the recipients. However, the levels of sHLA were not found to be a useful tool for monitoring rejection. Rejection was often associated with low levels of donor sHLA. The recent finding that antibodies can inhibit the detection of sHLA molecules might explain this discrepancy. In order to test this hypothesis, patient sera were screened for the presence of anti-HLA antibodies and the results were related to the donor-derived sHLA levels. Only in four out of 34 patients HLA Class I specific antibodies could explain the low sHLA levels during rejection. CONCLUSIONS: In heart transplantation increased donor-derived sHLA levels are not a suitable marker for rejection and that antibody formation can not explain these results. Therefore, monitoring rejection episodes on the basis of donor-derived soluble HLA molecules is not a realistic approach to decrease the number of biopsies after heart transplantation. PMID- 10834612 TI - Allograft tolerance induced by cyclophosphamide without prior inoculation of donor cells--immune suppression and redirection. AB - OBJECTIVES: To determine the possibility and cellular mechanism of inducing allograft tolerance by multiple injection of a lower dose of cyclophosphamide without prior infusion of donor cells. METHODS AND RESULTS: Heterotopic heart grafts were performed in MHC mismatched strain combinations (C57/B6 vs. BALB/c). Cyclophosphamide (40 mg/kg) was given intravenously on days 0, 2, 4 and 7 without prior infusion of donor cells. Long-term (> 100 days) allograft survival with normal histology was achieved. The long-term survivors accepted the donor skin grafts, but rejected the third-party skin grafts. Cyclophosphamide treatment initially led to profound lymphocytopenia, inhibition of spontaneous blastogenesis and low levels of lymphocyte proliferation response to both donor and third-party antigens. Ultimately, donor-specific tolerance occurred demonstrated by normal levels of peripheral lymphocytes, spontaneous blastogenesis and lymphocyte proliferation response to third-party antigens, and low levels of lymphocyte proliferation response to donor antigen. A switch of cytokines from IFNgamma dominant to IL-4 dominant, a low level of IgM and a high level of IgG1 were found in tolerant mice. CONCLUSIONS: Allograft tolerance can be induced by a short course of cyclophosphamide without prior donor cell inoculation. Tolerance induced is characterized initially by non-specific immunosuppression, which progresses to donor-specific hyporesponsiveness associated with the development of a Th2 dominant cytokine response. PMID- 10834613 TI - Effect of anti-LFA-1 monoclonal antibody on rat small bowel allograft survival and circulating leukocyte populations. AB - Anti-LFA-1 monoclonal antibodies (mAb) prolong graft survival in several animal models. This study assessed the effect of an anti-LFA-1 mAb (WT.1) on small bowel allograft rejection, circulating leukocyte subsets and in vivo target cell antigen blockade. Heterotopic small bowel transplantation was performed between PVG donor and DA recipient rats. Transplanted animals received 1 mg/kg per day WT.1 on days -1, 0 (day of transplantation) and 1. Three doses of WT.1 were also administered to a group of untransplanted animals to monitor circulating leukocyte populations and in vivo binding. WT.1 prolonged recipient survival from 7 to 14 days. Peripheral leukocyte counts increased more than twofold, primarily due to marked increases in both CD4+ and CD8+ lymphocytes. Approximately 85% of WT.1 binding sites on lymphocytes and monocytes were blocked/modulated after the course of therapy. WT.1 has marked effects on circulating leukocytes and target cell binding capacities and can affect the survival of rat small bowel transplant recipients. PMID- 10834614 TI - Optimization of adenoviral vector-mediated gene transfer to pulmonary arteries in newborn swine. AB - Efficient pulmonary vascular gene transfer in neonates would aid in understanding the pathophysiology of, and ultimately allow the development of specific therapies for, pulmonary vascular diseases. The purpose of this study was to optimize efficiency, and evaluate the duration, of catheter-based adenoviral vector-mediated pulmonary artery gene transfer in newborn pigs. An adenovirus vector encoding LacZ was infused via percutaneously placed catheters that were advanced to segmental pulmonary arteries under fluoroscopic guidance. Optimal viral dose and duration of expression were determined by histochemical evaluation of gene transfer efficiency 72 hr, 2 weeks, and 1 month after gene delivery. The effect of protamine on the efficiency of gene transfer was studied by assaying transgene protein in lung at 72 hr. The optimal viral dose was 2 x 10(10) plaque forming units (PFU). Seventy-two hours after infusion, expression predominated in medium-sized artery endothelial cells, 40% of which expressed beta-galactosidase. At 2 weeks, the distribution of expression had changed such that the majority of transduced cells were seen not in arteries but in gas exchange units of lung. No histochemical evidence of transgene expression was seen 1 month after virus infusion. The addition of protamine to virus infusate resulted in a fivefold increase in transgene protein product in lung tissue assayed 72 hr after gene transfer. Adenoviral vector-mediated gene transfer in neonatal swine results in high-efficiency transduction of arterial endothelial cells. However, the time course of gene transfer is not significantly prolonged compared with the adult. The addition of protamine results in a significant improvement in transduction efficiency, permitting lower doses of virus. PMID- 10834615 TI - Adenovirus-mediated transfer of caspase-8 augments cell death in gliomas: implication for gene therapy. AB - Caspase-8 is a member of the family of caspases, which are involved in the execution of apoptosis. To investigate whether caspase-8 can be used for gene therapy of gliomas, we transduced A-172 and U251 glioma cells with the caspase-8 gene via an adenoviral vector (Adv) controlled by the chicken beta-actin (CA) promoter (Advcaspase-8), and found that a similar level of caspase-8 protein induced A-172 cells to undergo necrotic cell death and U251 cells to undergo apoptotic cell death. Neither Bcl-XL nor Bcl-2, which play important roles in antiapoptotic mechanisms in gliomas, protected glioma cells from apoptosis induced by overexpression of caspase-8. Injection of Adv-caspase-8 suppressed the in vivo growth of U251 xenografts, in which apoptotic cell death remarkably increased as revealed by TUNEL analysis. Finally, we assessed whether gene therapy with a tissue-specific promoter, the myelin basic protein (MBP) promoter, is applicable to gliomas. Adv for caspase-8 controlled by the MBP promoter induced drastic apoptosis in U251 and U-373MG glioma cells, whereas it did not induce apoptosis in human endothelial cells, fibroblasts, and nerve growth factor treated PC12 cells. These results indicate that Adv for caspase-8 effectively induced cell death in gliomas, and that this approach may be a useful modality for gene therapy of gliomas. PMID- 10834616 TI - Reversal of pathology in the entire brain of mucopolysaccharidosis type VII mice after lentivirus-mediated gene transfer. AB - Gene transfer vectors derived from human immunodeficiency virus (HIV-1) efficiently transduce nondividing cells and remain stably integrated in their genome. Long-term expression of reporter genes has been documented after intracerebral injection of these vectors. Using a HIV-based vector, we looked for a reversal of brain damage in the beta-glucuronidase-deficient mucopolysaccharidosis type VII mouse, an animal model of human lysosomal storage diseases. The vector suspension was injected stereotactically in the brain of 10 week-old animals, an age at which storage lesions are patent in glia, perivascular cells, and neurons. Either a single intrastriatal injection or multiple injections in both cerebral hemispheres and in the cerebellum were performed. Local tolerance, enzyme delivery, and correction of storage lesions were investigated by comprehensive analysis of serial sections of the entire brain of mice killed 6 or 16 weeks postinjection. Histochemical staining detected enzyme activity in widely distributed areas, the size of which increased with time. Clearance of lysosomal storage extended far beyond enzyme-positive areas. In mice receiving multiple injections of the vector, complete correction or significant reduction of the pathology was observed in every section, suggesting disease regression in the entire brain. These results may have implications for the treatment of neurological symptoms in lysosomal storage diseases. PMID- 10834617 TI - Retrovirus-mediated gene transfer in primary T lymphocytes: influence of the transduction/selection process and of ex vivo expansion on the T cell receptor beta chain hypervariable region repertoire. AB - We have initiated a phase I/II clinical trial, involving the use of herpes simplex thymidine kinase gene (HS-tk)-expressing donor primary T cells, in order to modulate the graft-versus-host disease (GvHD) occurring after allogeneic hematopoietic stem cell transplantation. The preparation of gene-modified T cells (TkTCs) required a 12-day ex vivo culture comprising an initial OKT3 and IL-2 stimulation, a retrovirus-mediated transduction, and a 7-day selection step in the presence of G418 and IL-2. The low transduction efficiency as well as the culture conditions may significantly alter the diversity of the T cell repertoire. We therefore examined the T cell repertoire of HS-tk-expressing T cell samples from 11 different donors by the Immunoscope method. This method analyzes the hypervariable region of the T cell receptor beta chain (TCRBV) by amplifying the complementarity-determining region 3 (CDR3) and determining size diversity. In all examined samples (four of which were infused into patients), all TCRBV subfamilies were represented with, however, a significant skewing within a minority of subfamilies. Kinetic studies demonstrated that this skewing appeared between day 7 and day 12, with dates of appearance variable from one subfamily to another. In addition, the repertoire analysis of two different culture products, harvested and produced at different times from the same donors, suggested that some repertoire abnormalities could be donor specific. Quantitative analysis revealed no major modifications in gene usage, even in skewed TCRBV subfamilies, with a few clonal expansions concerning a limited number of TCRBV subfamilies. Importantly, identical abnormalities were found in control cells grown in parallel under similar conditions but not transduced or selected, thus demonstrating that these abnormalities were not related to the transduction or the selection process, but rather to the ex vivo culture. The initial stimulus used for T cell activation is a major source of TCRBV perturbation, since replacing the OKT3 + IL-2 stimulus by CD3 + CD28 monoclonal antibody-coated beads prevented the occurrence of alterations. Overall, the HS-tk expressing T cells used in our clinical trial exhibit limited TCR repertoire skewing that is not due to the transduction/selection procedure. However, future T cell gene transfer protocols for clinical trials should be designed to take into account or possibly prevent such T cell repertoire alterations. PMID- 10834618 TI - Assembly of human papillomavirus type 16 pseudovirions in Saccharomyces cerevisiae. AB - Studies of the encapsidation of papillomavirus (PV) DNA, and production of preparative amounts of PVs in vitro, have met with only limited success. To circumvent this problem we established a system in yeast to generate infectious HPV-16 pseudovirions. Saccharomyces cerevisiae strain 1699 was transformed with a construct to allow production of HPV-16 virus-like particles (VLPs). This strain was then transformed with a second construct (target plasmid), the same size as the HPV-16 genome and containing the HPV-16 upstream regulatory region (URR) and the HPV-16 E2 open reading frame. In addition, the target plasmid contained the green fluorescent protein gene to monitor delivery of the target plasmid into mammalian cells after infection. We conclude that this system allows HPV DNA encapsidation because (1) HPV-16 VLPs of two different types (heavy and light) were detected by CsCl gradient centrifugation, (2) DNase I-resistant DNA was detected by PCR/Southern blot analysis in fractions of CsCl gradients at a density corresponding to heavy VLPs, (3) in vitro infection of mammalian cells, including primary mouse splenocytes, with pseudovirions resulted in delivery of the reporter gene as demonstrated by FACS analysis for GFP expression, and (4) after injection of pseudovirions into mice, in vivo reporter gene expression was detected by confocal microscopy in sections of muscle tissue. We conclude that HPV-16 pseudovirions produced in yeast may be useful both for in vitro transduction and for gene delivery in vivo. PMID- 10834619 TI - Restoration of cognitive and motor functions by ciliary neurotrophic factor in a primate model of Huntington's disease. AB - Huntington's disease (HD) is an inherited disorder characterized by cognitive impairments, motor deficits, and progressive dementia. These symptoms result from progressive neurodegenerative changes mainly affecting the neostriatum. This pathology is fatal in 10 to 20 years and there is currently no treatment for HD. Early in the course of the disease, initial clinical manifestations are due to striatal neuronal dysfunction, which is later followed by massive neuronal death. A major therapeutic objective is therefore to reverse striatal dysfunction prior to cell death. Using a primate model reproducing the clinical features and the progressive neuronal degeneration typical of HD, we tested the therapeutic effects of direct intrastriatal infusion of ciliary neurotrophic factor (CNTF). To achieve a continuous delivery of CNTF over the full period of evaluation, we took advantage of the macroencapsulation technique. Baby hamster kidney (BHK) cells previously engineered to produce human CNTF were encapsulated into semipermeable membranes and implanted bilaterally into striata. We show here that intracerebral delivery of low doses of CNTF at the onset of symptoms not only protects neurons from degeneration but also restores neostriatal functions. CNTF treated primates recovered, in particular, cognitive and motor functions dependent on the anatomofunctional integrity of frontostriatal pathways that were distinctively altered in this HD model. These results support the hypothesis that CNTF infusion into the striatum of HD patients not only could block the degeneration of neurons but also alleviated motor and cognitive symptoms associated with persistent neuronal dysfunction. PMID- 10834620 TI - Retrovirus-mediated gene transfer into T cells: 95% transduction efficiency without further in vitro selection. AB - This study was designed to retrovirally transduce T cells by a protocol that would be simple, short, cost effective, applicable for clinical use, and efficient enough to avoid further selection of transduced T cells. Because retrovirally mediated infection is depending on the cell cycle, we first optimized the conditions for activating T cells in the presence of immobilized CD3 monoclonal antibodies and recombinant interleukin 2. Cell cycle analysis indicated that CD8+ and total T cells reach a maximum of cycling within 4 days whereas CD4+ T cells attain their maximum of cycling only by day 6. Taking into account these data, CD4+, CD8+, and total T cells were preactivated for 5 and 3 days, respectively, and then infected for 24 hr with supernatant containing retrovirus pseudotyped with gibbon-ape leukemia virus envelope, using a cell centrifugation protocol. Results show that approximately 95% of CD4+, CD8+, and total T cells can be transduced, this transduction efficiency being significantly higher than that obtained with amphotropic retrovirus vectors. Furthermore, under permanent growth stimulation, transduced T cells can be expanded approximately 1,000-fold in 4 weeks of culture with maintenance of transgene expression. However, Immunoscope analysis revealed alterations of T cell repertoire diversity after 2-3 weeks in culture that was not due to retroviral transduction per se. Overall, these data provide evidence that T cells can be transduced at levels that may alleviate the need for both further selection of transduced cells and in vitro expansion, thereby preserving the repertoire diversity of the transduced T cells to be reinfused. PMID- 10834621 TI - Gene therapy-directed osteogenesis: BMP-7-transduced human fibroblasts form bone in vivo. AB - An ex vivo gene therapy strategy was used to achieve localized skeletal regeneration in vivo. When an adenovirus vector engineered to express bone morphogenetic protein 7 transduced human gingival fibroblasts or rat dermal fibroblasts, these nonosteogenic tissues formed bone and supported the development of hematopoietic tissue when transplanted into immunocompromised mice. Transduced gingival fibroblasts formed marrow-containing ossicles in 100% of transplants after 1-2 weeks in vivo (n = 30). Immunostaining with murine and human-specific antisera raised against osteonectin and in situ hybridization of human-specific Alu genomic sequence demonstrated that the newly formed bone organ was a chimera of both the human donor and the mouse recipient cells. In experiments of greater clinical relevance, AdCMVBMP-7-transduced dermal fibroblasts repaired critical size skeletal defects in rat calvariae (n = 6). The results of this study suggest a bifunctional role of BMP-7-transduced fibroblasts. The transduced, nonosteogenic cells not only secreted biologically active BMP-7 in vitro and in vivo, but also differentiated into bone-forming cells in vivo. This model exploits the use of an easily biopsied, self regenerating tissue such as gingiva or skin and suggests that local regeneration of tissues by ex vivo gene therapy may not require that autogenous cells be cultured from the tissue that is to be regenerated. PMID- 10834622 TI - Binge eating disorder: a need for additional diagnostic criteria. AB - This study compares the core and associated features of binge eating disorder (BED) and bulimia nervosa (BN). One hundred twenty-nine adult females who were obese with BED (n = 51) or non-obese with BED (n = 32) or who had BN (n = 46) were compared using the Eating Disorder Examination-Questionnaire (EDE-Q). The BED groups were older and had a higher body mass index (BMI). The 3 groups were similar in binge frequency, but BN subjects (by definition) purged regularly. The groups differed by dietary restraint, even after controlling for BMI and age, such that the BN group had significantly higher dietary restraint than both BED groups. Cognitively, the 3 groups were similar in the intensity of dysfunctional attitudes regarding eating, weight, and shape. The BMI and age were not associated with these dysfunctional attitudes. Our findings suggest the importance of and the need to consider cognitive--as well as behavioral- diagnostic criteria for BED. PMID- 10834623 TI - Convergent and discriminant validity of DSM-IV axis II personality disorder criteria in adult outpatients with binge eating disorder. AB - The study objective was to evaluate the within-category cohesiveness and between category overlap of DSM-IV axis II personality disorders (PDs) in outpatients with binge eating disorder (BED). Seventy adult outpatients with BED were reliably administered the Diagnostic Interview for DSM-IV Personality Disorders (DIPD-IV). Within-category interrelatedness of the criteria was evaluated by Cronbach's alpha and mean intercriterion correlations (MICs). Between-category criterion overlap was evaluated by examining intercategory mean intercriterion correlations between all pairs of PDs (ICMICs). Cronbach's alpha was .64 to .93 (mean, .77), the MIC was .17 to .52 (mean, .34), and the ICMIC was .11 to .39 (mean, .28). Our findings indicate that in outpatients with BED, the DSM-IV PD criteria sets have convergent validity (acceptable alpha value and MIC). Some degree of discriminant validity also exists: criteria for most DSM-IV PDs correlate better with each other (MIC) than with criteria for other PDs (ICMIC). PMID- 10834624 TI - Insight and outcome in bipolar, unipolar, and anxiety disorders. AB - We performed a study to assess the relationship between impairment of insight and the long-term outcome in affective and anxiety disorders. Standardized insight assessments were made using the Scale to Assess Unawareness of Mental Disorder (SUMD) in 101 outpatients with psychiatric disorders, mostly affective and anxiety disorders, treated over 1 year in a university-based clinic. Outcome was prospectively assessed with the Clinical Global Impression (CGI) and Global Assessment of Functioning (GAF) rating scales. The mean follow-up period was 3.9 months. Initial impairment of insight did not correlate with poor outcome. However, improvement in insight correlated with good outcome, particularly in bipolar disorder type I (r = .56 to .67, P = .0005). Insight was similarly impaired in bipolar and unipolar major depressive disorders, and more so than in anxiety disorders (P = .002). An association between a lack of improvement in insight and a poor outcome, most significantly in bipolar disorder type I, was observed in this sample. We found a greater relative impairment of insight in mood versus anxiety disorders. PMID- 10834625 TI - Depersonalization in panic disorder: a clinical study. AB - Panic disorder (PD) has been hypothesized to be a heterogeneous entity, with distinct clinical subgroups. The presence of depersonalization during panic attacks may distinguish a specific subgroup of PD. We sought to analyze the differential features of a subgroup of PD patients with depersonalization. A total of 274 patients with PD were assessed and divided into 2 groups according to the presence or absence of depersonalization. The Structured Clinical Interview for DSM-III-R (SCID-UP-R) was used to assess PD and comorbid disorders. The clinical scales administered included the Hamilton Anxiety and Depression Rating Scale (HARS and HDRS), the Marks and Mathews Fears and Phobia Scale, Panic Associated Symptom Scale (PASS), and a panic attack symptoms inventory. A total of 66 patients (24.1%) exhibited depersonalization during the attacks. Patients with depersonalization appeared to be younger and had an earlier age at onset. PD was more severe in the depersonalization group (greater number of attacks, worse level of functioning, and higher scores on most self-rating scales). Also, depersonalization patients showed more comorbidity with specific phobia. Our results support the view that PD with depersonalization may be considered a distinct and more severe subcategory of PD. PMID- 10834626 TI - Predictors of remission in DSM hypochondriasis. AB - Although hypochondriasis is generally believed to be a chronic and refractory disorder, relatively little is known about its natural history and course. Based on a cognitive/perceptual model of hypochondriasis, we hypothesized that the disorder would be more chronic in patients who both amplify benign bodily symptoms and tend to attribute them to disease. Thirty-eight patients with DSM hypochondriasis were assessed with a structured, diagnostic interview and self report questionnaire. A logistic regression model containing sociodemographic characteristics and a 3-way interaction term composed of the tendency to amplify bodily sensations, the tendency to attribute common symptoms to disease, and somatization (all measured at inception) correctly classified the remission status of 81.6% of the patients at follow-up 4 years later. These results suggest that patients who somatize, who are amplifiers of bodily sensation, and those who tend to attribute ambiguous symptoms to disease have more chronic and more refractory hypochondriasis. It is the co-occurrence of these cognitive and perceptual characteristics, rather than their occurrence individually, which predicts the persistence of this disorder. PMID- 10834627 TI - First-episode schizophrenia: do grandiosity, disorganization, and acute initial development reduce duration of untreated psychosis? An exploratory naturalistic case study. AB - There is growing evidence for a relationship between the duration of untreated psychosis (DUP) and the prognosis. However, determinants of the DUP have been minimally explored, as is the detailed initial temporal unfolding of untreated psychosis. In this study, in-depth interviews with 19 first-episode DSM-IV schizophrenia patients and their relatives revealed detailed phenomenology of the initial prodrome and untreated psychosis. The findings suggest that a later prodrome onset (mean age in this sample, 20.5 years), a prodrome shorter than 2 years, acute initial psychosis development, the initial presence of grandiosity and/or disorganization, and a mild level of withdrawal all may reduce treatment delay. This set of characteristics might therefore represent built-in components of psychotic illnesses related to a shortened DUP, irrespective of early intervention efforts. In other words, the DUP could partly reflect these built-in aspects of psychosis. The rate of initial (untreated) psychosis development could contribute to broaden the prevailing mode of onset concept (presently defined as the duration of the initial prodrome) to comprise both the duration and content of the initial prodrome, as well as untreated psychosis, tentatively designated as the mode of initial psychosis development. PMID- 10834628 TI - Clinical predictors of discrepancy between self-ratings and examiner ratings for negative symptoms. AB - Little is known about the awareness of negative symptoms or its correlates. The aim of this study was to examine whether a number of clinical variables can predict the discrepancy between ratings of negative symptoms made by schizophrenic patients and by an examiner. This discrepancy could provide a measure for the awareness of negative symptoms. Eighty-six schizophrenic patients used a self-rating scale for negative symptoms with items derived from the Scale for the Assessment of Negative Symptoms (SANS). A psychiatrist assessed all patients using the SANS and other instruments, including the Present State Examination (PSE) item "insight into psychotic condition." Nurses assessed all patients using the Rehabilitation Evaluation Hall and Baker (REHAB), a scale for the measurement of psychiatric disability. All measurements were repeated after 2 months. A sensitive index for the underestimation of the severity of negative symptoms was developed, the discrepancy score. Multiple regression analysis was used to examine the predictability of discrepancy scores. Since scores for SANS items were used to calculate discrepancy scores, all regression analyses were performed with the SANS summary score as a covariate. The first step was to assess the independent contribution of each variable to the prediction of discrepancy scores. The second step was to examine the predictive quality of the 19 variables together. The variables themselves failed to make an independent contribution to the prediction of discrepancy scores at both assessments. REHAB scores, for instance, contributed to the prediction of discrepancy scores at the first assessment, but not at the second. The results of the second step showed that the best model for the prediction of discrepancy scores included the variables of depression (negative association) and anxiety (positive association). The absence of an association with the PSE item suggests that the awareness of negative symptoms is not related to insight into positive symptoms. An important limitation of our study is the arbitrary method of discrepancy quantification. PMID- 10834629 TI - Perceptions of schizophrenic patients and their therapists: application of the semantic differential technique to evaluate the treatment relationship. AB - This study is an attempt to evaluate the treatment relationship with schizophrenic patients by examining the patients' and their therapists' perceptions of themselves and each other, which are hypothesized to reflect features of the relationship. One hundred fifty-eight schizophrenic patients and 11 psychiatrists who maintained a supportive relationship with the patients as a therapist estimated their perceptions using the semantic differential (SD) technique with 17 adjective pairs. Eight composite scales with sufficient internal consistency were constructed from the estimations. The interrelationship among the perceptual elements, which was represented by correlation analysis of the composite scale scores, seemed consistent with our clinical experience. A factor-analytic study of the scales yielded 3 orthogonal factors that could be assumed to characterize the treatment relationship. The patient-therapist cooperation factor indicated the degree of trust between the two participants, supposedly the affective or relational aspect of the therapeutic alliance. The therapist passivity factor reflects the therapist's passive role-taking and the clinical stability of the patient. The patient strength factor was related to the condition-related and characterological strength of the patient. It is demonstrated that the estimations performed by patients and therapists are valid and useful for evaluation of the treatment relationship in the current status. PMID- 10834630 TI - Patterns of covariation of DSM-IV personality disorders in a mixed psychiatric sample. AB - The covariation patterns of DSM-IV personality disorders (PDs) were studied in 431 consecutively admitted psychiatric patients. The co-occurrence rate was greater than 50% for all DSM-IV PDs. Both bivariate association tests and loglinear models showed distinct significant covariation patterns among PDs which were stable across confounder strata. DSM-IV PD clusters were not replicated, with the exception of cluster A. Principal-component analysis (PCA) showed the presence of 3 latent dimensions, thus explaining the DSM-IV PD covariation patterns. These results seem to stress the inadequacy of the DSM-IV categorical model of PD assessment. The need for a reduction of axis II categories and the inclusion of a dimensional model in the diagnostic assessment of DSM-IV PDs are discussed. PMID- 10834631 TI - Frequency of dissociative disorders among psychiatric outpatients in Turkey. AB - The aim of this study was to determine the frequency of dissociative disorders among psychiatric outpatients in Turkey. One hundred fifty consecutive outpatients admitted to the psychiatry clinic of a university hospital were screened with the Dissociative Experiences Scale (DES). Twenty-three patients (15.3%) with a DES score greater than 30 and a comparison group selected from the same outpatient population who scored less than 10 on the scale were then interviewed with the Dissociative Disorders Interview Schedule (DDIS) in a blind fashion. According to the DDIS, 18 patients (12.0%) received a diagnosis of dissociative disorder; 83.3% (n = 15) of the dissociative patients reported neglect, 72.2% (n = 13) emotional abuse, 50.0% (n = 9) physical abuse, and 27.8% (n = 5) sexual abuse during childhood. Dissociative disorders are not rare among psychiatric outpatients. Self-rating instruments and structured interviews can be used successfully for screening dissociative disorders, which are usually underrecognized. Neglect was the most frequently reported type of childhood trauma, suggesting the importance of other childhood experiences in addition to sexual and/or physical abuse in the development of dissociative psychopathology. PMID- 10834632 TI - The dissociative experiences of borderline patients. AB - The study objective was to assess the severity and quality of dissociative experiences reported by borderline patients. Two hundred ninety criteria-defined borderline patients and 72 axis II controls completed the Dissociative Experiences Scale (DES), a 28-item self-report measure with demonstrated reliability and validity. Thirty-two percent of borderline patients had a low level of dissociation, 42% a moderate level, and 26% a high level similar to that reported by patients meeting criteria for posttraumatic stress disorder (PTSD) or dissociative disorders. The controls had a significantly different distribution of overall DES scores: 71% reported a low level of dissociation, 26% reported a moderate level, and only 3% reported a high level. In addition, borderline patients had a significantly higher score than the controls on 21 of 28 DES items and a significantly higher overall DES score, as well as the score on the 3 factors that have been found to underlie the DES, absorption, amnesia, and depersonalization. The results of this study suggest that the severity of dissociation experienced by borderline patients is more heterogeneous than previously reported. They also suggest that borderline patients have a wider range of dissociative experiences than are commonly recognized, including experiences of absorption and amnesia, as well as experiences of depersonalization. PMID- 10834633 TI - Nerve surgery as it is now and as it may be. PMID- 10834634 TI - What percentage of surgically clipped intracranial aneurysms have residual necks? AB - OBJECTIVE: To determine the angiographically proven rate and persistence of occlusion of intracranial aneurysms after surgical clipping as reported in the literature. This should establish a basis for comparing surgery with new endovascular methods of treatment. METHODS: We reviewed the literature published during the period from 1979 through 1999, dividing the articles into two groups. The first group of articles reported patients undergoing surgical treatment with immediate postoperative angiography. The second group of articles documented symptomatic recurrence or regrowth of aneurysms that were surgically treated previously. The data from these articles are presented for analysis. RESULTS: During the period 1979 to 1999, six series of patients undergoing surgical treatment of aneurysms with immediate postoperative angiography were reported. These reported series comprised a total of 1,397 patients, of whom 1,370 underwent postoperative angiography demonstrating 1,569 clipped aneurysms. Residual filling was found in 82 aneurysms (5.2%) on postoperative angiography. Of the 1,370 patients, only 124 patients with 169 aneurysms were reported to have had any long-term angiographic follow-up. The second group consisted of 226 patients representing six reported groups of patients, who either presented up to 24 years after aneurysm clipping with recurrent symptoms of hemorrhage or mass effect, or had important findings on intraoperative and postoperative angiograms. CONCLUSION: The lack of information regarding both the frequency of residual filling or regrowth and long-term angiographic follow-up of patients with surgically treated aneurysms makes meaningful comparison between surgical treatments and new treatment methods for intracranial aneurysms difficult or impossible. Detailed analysis with high-quality angiography should be performed to determine the success of surgical treatment. PMID- 10834635 TI - Clinical features of intracranial aneurysms in siblings. AB - OBJECTIVE: Among the family members of patients with aneurysmal subarachnoid hemorrhage (SAH), siblings have been documented to be at high risk of SAH and to have a high prevalence of unruptured aneurysms. We studied the distinctive features of aneurysms in siblings and attempted to determine the risk of rupture. METHODS: We analyzed detailed data on 159 patients with siblings who had ruptured and unruptured aneurysms in 77 families from throughout Japan. RESULTS: Seventy three percent of the patients were female, and the mean age at the time of rupture was 55.6 years. In 39 families, two or more siblings had SAH. Eighty of 107 patients with ruptured aneurysms and 28 of 52 with unruptured aneurysms had a family history of SAH in siblings (P = 0.0082). Multiple and mirror-image aneurysms were found in 42 and 21 patients, respectively. Among 218 aneurysms, middle cerebral artery aneurysms were the most common type (43%). Anterior communicating artery aneurysms were underrepresented (15%). There were significantly more ruptured than unruptured anterior communicating artery aneurysms, compared with other aneurysms (P = 0.01). CONCLUSION: The clinical features of aneurysms in siblings in this population agreed well with those reported for familial intracranial aneurysms and SAH, except for the age at the time of rupture. It is suggested that the risk of rupture is greater when patients with unruptured aneurysms have siblings with aneurysmal SAH and/or anterior communicating artery aneurysms. PMID- 10834636 TI - A grading scale to predict outcomes after intra-arterial thrombolysis for stroke complicated by contrast extravasation. AB - OBJECTIVE: Contrast extravasation after intra-arterial thrombolysis for stroke occurs frequently and is identifiable on a computed tomographic (CT) scan, but it is often unrecognized or misdiagnosed. Few articles describing this phenomenon have been published. The clinical outcomes of patients after contrast extravasation are poorly understood. We designed a grading system to predict outcomes after contrast extravasation and tested the grading scale prospectively. METHODS: We studied 27 patients who had contrast extravasation exhibited on a CT scan immediately after intra-arterial thrombolysis. The National Institutes of Health Stroke Scale was used to quantify neurological examinations preoperatively, postoperatively, and at follow-up an average of 3 months later. A grading scale from 0 to 10 was developed from a retrospective analysis of the first 18 patients using odds ratios and Fisher's exact test. The grading system was then applied prospectively to the next 9 consecutive patients. RESULTS: Six components of the grading system were weighted approximately proportional to corresponding odds ratios: 1) incomplete recanalization (3 points), 2) prolonged angiographic blush (2 points), 3) hyperdensity greater than 150 Hounsfield units (2 points), 4) lesion volume greater than 50 cc exhibited on a CT scan (1 point), 5) lesion in eloquent parenchyma (1 point), and 6) hypodensity demonstrated on an immediate postoperative CT scan (1 point). The contrast extravasation grades for each outcome category (excellent, fair, poor, died) increased in stepwise fashion. There was a direct linear correlation between the assigned grade and National Institutes of Health Stroke Scale score improvement at follow-up. CONCLUSION: This grading system should prove useful as a preliminary guide for predicting outcomes of patients with contrast extravasation after intra-arterial thrombolysis for stroke. Further analysis in a large cohort of prospective patients is necessary to ensure extensibility. PMID- 10834637 TI - Abciximab as an adjunct to high-risk carotid or vertebrobasilar angioplasty: preliminary experience. AB - OBJECTIVE: Abciximab, a platelet glycoprotein IIb/IIIa receptor inhibitor, has been shown to reduce the risk of ischemic events associated with coronary intervention. However, its role in neurointerventional procedures needs to be defined. We prospectively evaluated our initial experience with the use of abciximab in a series of high-risk patients undergoing carotid, basilar, or vertebral artery angioplasty. METHODS: Patients were given an intravenous abciximab bolus (0.25 mg/kg), followed by infusion (10 microg/min) for a period ranging from 12 to 24 hours, as an adjunct to angioplasty in 20 procedures (19 patients). These patients were considered to be at high risk for thromboembolic events because of recent ischemic symptoms and/or complex lesion morphology. Before, immediately after, and 24 hours after the procedure, each patient was evaluated by a neurologist for the presence of new neurological deficits. Any bleeding or other complications during hospitalization were also recorded. Bleeding was defined as major (hemoglobin decrease >5 g/dl), minor (hemoglobin decrease 3-5 g/dl), or insignificant. RESULTS: Angioplasty was performed in the internal carotid artery (n = 13), vertebral artery (n = 4), or basilar artery (n = 2). Stents were placed across 13 lesions. In one patient, angioplasty could not be performed owing to technical difficulties; however, abciximab was administered because of extensive lesion manipulation. Intraprocedural heparin was given in 19 procedures (35-86 U/kg intravenously) and partially reversed in 6 procedures. Low dose intra-arterial thrombolytic agents were administered in seven patients before the lesion was crossed. Two patients experienced transient neurological deficits either during (n = 1) or immediately after (n = 1) the procedure. Another patient had complete occlusion of the right vertebral artery after angioplasty with complete recanalization after 24 hours of abciximab infusion. Major or minor bleeding was not observed in any patient. Insignificant bleeding was observed in eight patients. Thrombocytopenia was observed in one patient who received concomitant administration of intravenous heparin and abciximab infusion. CONCLUSION: We observed a low frequency of neurological events in high risk patients undergoing angioplasty with or without stent placement. Abciximab seems to be a relatively safe adjunct for carotid or vertebrobasilar endovascular intervention either alone or in combination with low-dose thrombolytics. Partial reversal of intraprocedural heparin should be considered to reduce the risk of postprocedural bleeding. PMID- 10834638 TI - Factors associated with reintegration to normal living after subarachnoid hemorrhage. AB - OBJECTIVE: Recent reports have suggested improvement in the last decade in global outcome measures after subarachnoid hemorrhage (SAH), particularly in patients presenting in good initial neurological status. We used a standardized self report instrument, the Reintegration to Normal Living (RNL) Index, to assess a patient-based quality of life measure and a self-report of work status. We tested the hypothesis that several patient-based factors were related to these outcomes, including depressive symptoms, physical disability, age, and initial Hunt and Hess grade. Using these data, we report the total management morbidity and mortality at 1 to 5 years after SAH for patients initially presenting in good neurological condition. METHODS: The study population consisted of 246 consecutive patients admitted to our tertiary care center with aneurysmal SAH in good neurological condition (Hunt and Hess Grades I-III). Patients underwent either surgical (92%), endovascular (7%), or medical (1 %) management of aneurysmal SAH. Eighty-three percent of surviving patients completed a written or telephone questionnaire incorporating the Barthel Index, the Zung Self-rating Depression Scale, the RNL Index, and a work status assessment. RESULTS: An aneurysm-related mortality rate of 6% was observed in the patient population. Fifty-five percent of patients reported a complete reintegration into their normal living situation, as measured by the RNL Index. Sixty-seven percent of previously full-time workers returned to a full-time status. Thirty-six percent of patients reported depressive symptoms, and 23% of patients reported physical disability. In a multivariate model, the two factors that contributed most to an impairment of reintegration were depression and physical disability, whereas a failure to return to work was related to older age and a higher-grade initial neurological status. CONCLUSION: In this series of patients undergoing multimodality management of lower-grade SAH, more than one-half of patients subsequently reported a normal reintegration into their social situation, as assessed by the RNL Index. Standardized assessments of cofactors associated with impaired reintegration revealed that depressive symptoms and physical disability played a strong role in overall reintegration. Standardized assessments, such as the RNL Index, offer the potential for improved comparison of different treatment regimens and specific therapeutic targeting of factors, such as depression, which contribute to decreased quality of life. PMID- 10834639 TI - The use of stents in the management of neurovascular disease: a review of historical and present status. AB - In the mid-1960s, radiologists began experimenting with stents for use in the peripheral vasculature in the hope of treating vascular insufficiency resulting from vessel stenosis in a nonsurgical manner. The 1990s saw stents move into the neurovascular arena for the management of a variety of disease processes, including arterial and venous sinus stenosis, arterial dissection, arterial aneurysms, and arteriovenous fistulae. This article reviews the current status of stenting in regard to the management of neurovascular maladies. PMID- 10834640 TI - Prevention and treatment of thromboembolic and ischemic complications associated with endovascular procedures: Part I--Pathophysiological and pharmacological features. AB - Thromboembolic and ischemic complications frequently occur during and after endovascular procedures, because of associated arterial injury and the thrombogenic characteristics of arterial catheters, contrast agents, and implanted devices such as coils and stents. Platelet adhesion, activation, and aggregation occurring at the site of arterial injury are mediated by local factors, including thromboxane A2 (inhibited by aspirin) and adenosine diphosphate (inhibited by ticlopidine and clopidogrel). Concomitantly, thrombin is formed by serial activation of clotting factors via contact with subendothelial tissue factor. Thrombin cleaves fibrinogen into fibrin. Thrombin activation is indirectly blocked by heparin and its analogs. However, after thrombin is clot-bound (with fibrin), it is relatively protected from heparin and is effectively blocked only by direct thrombin inhibitors (hirudin and its analogs). The final common pathway in clot formation is the binding of fibrinogen to platelets via platelet glycoprotein IIb/IIIa receptors, which is inhibited by antibodies to platelet IIb/IIIa receptors. New treatment modalities, such as the use of direct thrombin inhibitors and antibodies to platelet glycoprotein IIb/IIIa, seem to be more effective for prophylaxis and treatment than conventional anticoagulation and antiplatelet therapies. PMID- 10834641 TI - Prevention and treatment of thromboembolic and ischemic complications associated with endovascular procedures: Part II--Clinical aspects and recommendations. AB - We reviewed the incidence, risk factors, and clinical features of thromboembolic and ischemic events associated with diagnostic cerebral angiography, endovascular treatment of aneurysms using coils or balloons, angioplasty and stent placement to treat extracranial carotid artery stenosis, and embolization of arteriovenous malformations using glue or other embolic agents. We performed a cumulative analysis to determine the frequency and characteristics of these events and a subset analysis (whenever possible) to determine the benefits of various strategies for complication avoidance. Of the 1,547 patients who underwent Guglielmi detachable coil treatment, thromboembolic events were observed for 127 (8.2%), consisting of asymptomatic events for 12 patients, transient ischemic attacks for 29, and strokes for 86. The outcomes for the 86 patients with strokes were categorized as full recovery for 15, good recovery for 27, partial recovery for 19, no recovery for 11, death for 12, and undetermined outcome for 2. Of the 834 patients who underwent carotid angioplasty and stent placement, thromboembolic events were observed for 73 (8.8%), consisting of transient ischemic attacks for 26 patients and strokes for 47. The outcomes for the patients with strokes were categorized as full recovery for 20, good recovery for 15, partial recovery for 6, no recovery for 2, and death for 4. High rates of thromboembolic events were also observed with balloon occlusion of aneurysms (11%) or parent arteries (19%) and carotid angioplasty alone (5.9%). Arteriovenous malformation embolization was associated with an ischemic event/procedure rate of 9.4%. High rates of thromboembolic and ischemic complications, with subsequent morbidity and death, are associated with most endovascular procedures. Further research and the formulation of standard preventive guidelines may help to reduce these risks and improve the overall success of these procedures. PMID- 10834642 TI - Advanced surgical approach for selective amygdalohippocampectomy through neuronavigation. AB - OBJECTIVE: Selective removal of the mesiobasal temporal structures through the transsylvian approach was introduced by Yasargil and Wieser in 1982. This alternative to standard temporal lobectomy provides excellent outcomes for seizure control. Basic actions in the transsylvian fissure exposure mainly serve to orient the surgeon, and they carry the risk of vasospasm and vessel damage. The aim of our study was to reduce landmark-guided surgery steps through neuronavigation. METHODS: During a 14-month period, 16 selective amygdalohippocampectomies were performed with the aid of the SMN (Carl Zeiss, Inc., Thornwood, NY) or StealthStation (Sofamor Danek, Memphis, TN) optically guided systems. We added safety procedures to the operation (including intraoperative rereferencing, obtaining additional bony reference points before craniotomy, performing a small craniotomy and making an accurate dural incision, and using contrast medium for vessel visualization) to develop a method that relies on navigational systems without further orientation by anatomic landmarks. RESULTS: Originally, performing an amygdalohippocampectomy required exposing the sylvian fissure from the carotid bifurcation to 2 cm beyond the middle cerebral artery bifurcation, which exposed one-third of the insula. By determining the entry point at the limen insulae and the target at the tip of the temporal horn, the mandatory extent of the opening to the sylvian fissure can be projected. Therefore, the exposure of the fissure can be limited to exactly the extent required for the transventricular approach through the uncinate fasciculus. CONCLUSION: Computer-assisted surgery is an effective tool in eliminating the exposure of anatomic landmarks in selective amygdalohippocampectomy. This modification combines the precision of targeting with minimal cortical and vessel traumatization. PMID- 10834643 TI - Effects of posterior fossa decompression with and without duraplasty on Chiari malformation-associated hydromyelia. AB - OBJECTIVE: The optimal surgical treatment of Chiari malformation is unclear, especially in patients with hydromyelia. Various surgical approaches have included suboccipital craniectomy, syringostomy, obex plugging, syringosubarachnoid shunting, and fourth ventriculosubarachnoid shunting. The purpose of this study is to differentiate extradural and intradural approaches in the treatment of Chiari I malformation. METHODS: We reviewed the medical records and magnetic resonance imaging (MRI) scans of 34 surgical corrections' of Chiari malformation performed at our institution from 1988 to 1998. The age and sex of the patient, the presence of hydromyelia, the type of surgery (duraplasty or nonduraplasty), and the clinical outcome were determined. RESULTS: Eleven patients underwent posterior fossa decompression (PFD) and C1 laminectomy without duraplasty. Eight (73%) of these patients had an improvement in symptoms. Seven of the 11 patients had hydromyelia. Of the six patients who underwent follow-up MRI, three (50%) had a decrease in the size of the hydromyelia, and all three had clinical improvement. We also noted a morphometric increase in posterior fossa volume on postoperative MRI scans in these three patients, which was not observed in those without improvement. Two of the three patients whose hydromyelia did not decrease on follow-up MRI scans worsened clinically, and one underwent a reoperation with duraplasty. Twenty-three patients underwent combined PFD, C1 laminectomy, and duraplasty. Twenty (87%) of these patients had improvement. Twelve of the patients who underwent duraplasty had hydromyelia; nine underwent follow-up MRI. All nine of these patients (100%) had a decrease in the cavity size, including eight with clinical improvement. There were 10 minor complications (seroma, 4; superficial infection, 3; cerebrospinal fluid leak, 2; aseptic meningitis and occipital nerve pain, 1) when the dura was opened, compared with one superficial wound infection that resolved in patients who underwent PFD only. CONCLUSION: PFD, C1 laminectomy, and duraplasty for the treatment of Chiari I malformation may lead to a more reliable reduction in the volume of concomitant hydromyelia, compared with PFD and C1 laminectomy alone. However, there seems to be a subset of patients whose symptoms will resolve and whose hydromyelic cavity will decrease with the removal of bone only. These patients seem to undergo a volumetric increase in the posterior fossa. Further studies are needed to better characterize these patients, to determine which patients with Chiari I malformation are better served with bony decompression only, and which will require duraplasty to resolve their hydromyelia. PMID- 10834644 TI - Dural repair using acellular human dermis: experience with 200 cases: technique assessment. AB - OBJECTIVE: Many craniotomies require a watertight dural closure. When primary dural repair is not possible, a graft is necessary. Autograft material is not always easily accessible or available, necessitating the use of other material. We performed 200 craniotomies using an acellular human dermal graft (AlloDerm; LifeCell Corp., The Woodlands, TX) to determine its suitability as a dural substitute. METHODS: From June 1996 through March 1998, all patients at Allegheny General Hospital who required a dural substitute graft and in whom autograft harvest was impractical or impossible received the acellular dermal autograft. The running suture technique was used to form a watertight seal. RESULTS: After follow-up for a minimum of 1 year, seven patients have required subsequent surgery. Three patients developed cerebrospinal fluid leaks that were repaired without removing the dermal graft. Four patients developed wound infections that required debridement. In each patient, the graft seemed to be uninvolved in the infectious process and was left in place. The patients were administered antibiotics postoperatively, and there have been no recurrent infections. No adhesion formation or scarring was noted around or underneath the graft in any patient. CONCLUSION: AlloDerm is a reasonable alternative to the available dural graft materials. Its handling characteristics are similar to those of dura, it is biologically inert, and it does not produce adhesion formation. PMID- 10834646 TI - Case problems conference: thoracic spinal cord hernia. PMID- 10834645 TI - Balloon-assist technique for endovascular coil embolization of geometrically difficult intracranial aneurysms. AB - OBJECTIVE: The balloon-assist or neck-remodeling technique is an adjunctive method devised for the endovascular coil embolization of aneurysms characterized by a wide neck or unfavorable geometric features. Since its initial description, there have been few data to corroborate its utility, efficacy, and safety in aneurysm embolization. METHODS: Twenty patients (19 female patients and 1 male patient) with 22 aneurysms (19 unruptured aneurysms and 3 ruptured aneurysms) underwent balloon-assisted coil embolization. The balloon-assist technique was performed in the same treatment session after conventional coil embolization had failed in 55% of cases (12 of 22 cases) and was the primary treatment in 45% of cases. The majority of aneurysms were located in the supraclinoid carotid artery (13 paraophthalmic and 3 superior hypophyseal aneurysms). The mean angiographic measurements included a fundus of 8.7 +/- 3.7 mm, a neck of 5.3 +/- 2.2 mm, and a comparatively unfavorable fundus/neck ratio of 1.33 +/- 0.23. RESULTS: Technical success was achieved in 77% of cases (17 of 22). The rate of aneurysm obliteration at the end of the procedures was 97 +/- 3.8%. Angiographic follow-up data (mean follow-up period, 10.3 mo) obtained for 89% of the treated aneurysms (15 of 17) confirmed stable mean occlusion of 97.8 +/- 3.8%. Technical complications included two cases of asymptomatic distal vessel thromboembolism, which resolved angiographically within 24 hours, and one case of intraprocedural rupture of an arteriovenous malformation-related feeder artery aneurysm, which resulted in no neurological deficits and required no further treatment (transient complication rate, 13.6%; 3 of 22 cases). There were no deaths and no procedure related 30-day or permanent morbidity. CONCLUSION: The balloon-assist method of coil embolization is characterized by promising intermediate-term angiographic and clinical outcomes and acceptable morbidity and mortality rates. Although this adjunctive method requires the use of an additional microcatheter and consequently involves a higher level of technical complexity, it extends the range of aneurysms that can be successfully treated with electrolytically detachable coils via an endovascular approach. PMID- 10834647 TI - Unilateral upper and lower subtotal maxillectomy approaches to the cranial base: microsurgical anatomy. AB - OBJECTIVE: The relationship of the maxilla, with its thin walls, to the nasal and oral cavities, the orbit, and the infratemporal and pterygopalatine fossae makes it a suitable route for accessing lesions involving both the central and lateral cranial base. In this study, we compared the surgical anatomy and exposure obtained by two unilateral transmaxillary approaches, one directed through an upper subtotal maxillectomy, and the other through a lower subtotal maxillectomy. METHODS: Cadaveric specimens examined, with 3 to 40x magnification, provided the material for this study. RESULTS: Both upper and lower maxillectomy approaches open a surgical field extending from the ipsilateral internal carotid artery to the contralateral Eustachian tube; however, they differ in the direction of the access and the areas exposed. The lower maxillectomy opens a combination of the transmaxillary, transnasal, and transoral routes to extra- and intradural lesions of the central cranial base. Performing additional osteotomies of the mandibular coronoid process and the sphenoid pterygoid process provides anterolateral access to the lateral cranial base, including the pterygopalatine and infratemporal fossae, and the parapharyngeal space. The upper maxillectomy opens the transmaxillary and transnasal routes to the central cranial base but not the transoral route. The structures exposed in the lateral cranial base, after removing the coronoid and pterygoid processes, include the pterygopalatine and infratemporal fossae and the parapharyngeal space. Exposure can be extended by a frontotemporal craniotomy, which provides access to the anterior and middle cranial fossae and the basal cisterns. CONCLUSION: The upper and lower subtotal maxillectomy approaches provide wide but differing access to large parts of the central and lateral cranial base depending on the site of the osteotomies. PMID- 10834648 TI - Ultrastructural pathological features of cerebrovascular malformations: a preliminary report. AB - OBJECTIVE: Although cerebrovascular malformations have been characterized histologically, a systematic examination of such lesions by transmission electron microscopy has not been previously published. In this preliminary study, we describe the ultrastructural pathological features of cerebral arteriovenous malformations (AVMs) and cavernous malformations (CMs). METHODS: Using transmission electron microscopy, we examined three CMs and three AVMs microsurgically harvested from patients, for conventional indications. Normal control cerebral tissue was obtained from two patients undergoing surgery for epilepsy. Specific attention was directed at components of the vascular wall, endothelial cell (EC) morphology, intercellular tight junctions, and the subendothelial layer. RESULTS: In embolized AVM vessels, ECs were disrupted, with preservation of the underlying subendothelial vessel wall. Nidal vessel walls of AVMs showed disorganized collagen bundles. In CM specimens, ECs lined attenuated cavern walls that were composed of an amorphous material lacking organized collagen. Peripheral to the CMs, capillaries were often surrounded by rings of hemosiderin. Tight junctions between ECs were present in AVMs and control specimens, but substantial inter-EC gaps were found in CMs. Subendothelial smooth muscle cells were present in AVM and control specimens, but they were sparse or poorly characterized in CMs. CONCLUSION: Surgically resected vascular malformations demonstrate abnormal ultrastructural pathological features. The preoperative embolization of AVMs results in EC denudation, with preservation of vessel wall structural integrity. The thin walls of CMs, lacking significant subendothelial support, along with the rarity of intact tight junctions between ECs, may contribute to the known propensity of CMs for recurrent microhemorrhage. PMID- 10834649 TI - Stereotactic injection of adenoviral vectors that target gene expression to specific pituitary cell types: implications for gene therapy. AB - OBJECTIVE: Gene therapy is a potentially useful strategy for the treatment of pituitary adenomas or hormone deficiency disorders. We investigated the feasibility of targeting gene expression to specific pituitary cell types in vivo, using a combination of stereotactic injection and adenoviral vectors that carry pituitary-specific promoters. METHODS: Recombinant adenoviruses containing the human growth hormone promoter (AdGHGal) or the human glycoprotein hormone alpha-subunit promoter (AdalphaGal) were used to drive expression of the beta galactosidase gene. The expression of beta-galactosidase activity in the pituitary was analyzed after the administration of recombinant adenoviruses via the peripheral vein or the carotid artery, or by stereotactic injection into the rat pituitary. Double-label histology was used to evaluate cell-type expression in the pituitary. RESULTS: Intravascular injection of AdGHGal or AdalphaGal failed to deliver the marker gene to the pituitary. However, direct stereotactic injection of recombinant adenoviral vectors into the pituitary achieved a high level of transgene expression. In addition, immunohistochemical staining revealed selective expression of the AdGHGal or AdalphaGal transgenes in pituitary cells that normally produce the respective hormones. CONCLUSION: These findings indicate that adenoviral vectors carrying pituitary gland-specific promoters may be useful for targeted gene therapy of pituitary diseases. However, because of low transduction after peripheral administration, stereotactic injection or local administration of viruses at the time of pituitary surgery is probably required for efficient gene expression. PMID- 10834650 TI - Tightness of duraplasty in rabbits: a comparative study. AB - OBJECTIVE: The purpose of this study was to test, in rabbits, the tightness of seven dural substitution materials commonly used in neurosurgery, i.e., Lyodura (B. Braun Melsungen AG, Melsungen, Germany), Tutoplast dura (Tutogen Medical, Inc., Parsippany, NJ), Tutoplast fascia lata (Tutogen Medical, Inc.), autologous periosteum, Neuropatch (B. Braun Melsungen AG), Dacron (E.I. du Pont de Nemours and Co., Wilmington, DE), and Ethisorb (Ethicon, Inc., Somerville, NJ). METHODS: Duraplasties were performed with sutures alone or were additionally fixed with fibrin glue. Leakage pressures were assessed by infusion of artificial cerebrospinal fluid, containing sodium fluorescein, into the cisterna magna and detection of fluorescence using a charge-coupled display camera with background substraction, 3 days, 3 weeks, or 3 months after surgery. RESULTS: Three days after implantation, the mean tightness values of duraplasties with Lyodura or Neuropatch were significantly higher (P = 0.007) than the values for the other substitutes. A significant improvement of tightness with increasing implantation time could be demonstrated for autologous periosteum (P = 0.0063). Improvement of tightness with the use of fibrin glue could be proven only for the heterologous grafts (P = 0.0071). The tightness values for Neuropatch fixed only with sutures were similar to those for the best heterologous substitutes implanted with additional fibrin glue. Lyodura, Tutoplast dura, and Neuropatch demonstrated favorable implantation characteristics; they were thin, flexible, and easily suturable. Neither adhesions to the brain nor space-occupying scars were noted. CONCLUSION: These results confirm the excellent suitability of Lyodura and Neuropatch for dural substitution. PMID- 10834651 TI - Inhibition of peridural fibrosis after laminectomy using low-dose external beam radiation in a dog model. AB - OBJECTIVE: Clinical studies have demonstrated a significant association between the presence of extensive postlumbar discectomy peridural scar formation and the recurrence of low back and radicular pain. Low-dose perioperative radiation therapy has previously been demonstrated to inhibit peridural fibrosis after laminectomy in a rat model. The current study was designed to measure the effect of low-dose radiation on postlaminectomy peridural fibrosis development in a larger animal model. METHODS: Three dogs underwent a total of 12 lumbar hemilaminectomies. For each animal, two levels received 1) external beam radiation 24 hours before surgery, 2) surgery alone, or 3) radiation alone. Radiation was administered in a single fraction of 700 cGy using computed tomographic guidance for dosimetry planning. The isodose distribution was such that the dose conformed to the posterior epidural space with minimal exit dose to normal tissue. Port films were used to confirm the correct levels. Gadolinium enhanced magnetic resonance imaging (MRI) of the lumbar spines was obtained before the animals were killed 12 weeks after surgery. The spines were harvested, and axial sections through the laminectomy defect were stained with hematoxylin and eosin and Masson's trichrome. All specimens were evaluated for extent of fibrosis along the dura, density of fibrosis, nerve root entrapment, and sublaminar fibrosis. RESULTS: There were no complications from the surgery, and no new neurological deficits were noted. There was a statistically significant difference between the irradiated and nonirradiated groups regarding the extent of fibrosis (P = 0.001) and the density of fibroblasts (P = 0.001). There was also a marked difference in nerve root entrapment (P = 0.182) and the presence of sublaminar fibrosis (P = 0.061) between the treatment and control groups. MRI revealed less gadolinium enhancement at the irradiated levels compared with the nonirradiated levels, confirming the usefulness of MRI in predicting the degree of epidural fibrosis. CONCLUSION: Low-dose external beam radiation therapy administered 24 hours before laminectomy in a dog model significantly decreased the extent and density of peridural fibrosis as well as nerve root entrapment and sublaminar fibrosis. This treatment strategy may be efficacious in patients with recurrent radicular pain after lumbar discectomy that is thought to be secondary to peridural fibrosis on the basis of gadolinium-enhanced MRI studies, and who might benefit from reoperation for nerve root decompression. PMID- 10834652 TI - Program requirements for residency/fellowship education in neuroendovascular surgery/interventional neuroradiology: special report on graduate medical education: a joint statement by the American Society of Interventional and Therapeutic Neuroradiology, Congress of Neurological Surgeons and American Association of Neurological Surgeons, American Society of Neuroradiology. AB - Neuroendovascular surgery/interventional neuroradiology is a relatively new subspecialty that has been evolving since the mid-1970s. During the last 2 decades, significant advances have been made in this field of minimally invasive therapy for the treatment of intracranial cerebral aneurysms; acute stroke therapy intervention; cerebral arteriovenous malformations; carotid-cavernous sinus fistulae; head, neck, and spinal cord vascular lesions; and other complex cerebrovascular diseases. Advanced postresidency fellowship programs have now been established in North America, Europe, and Japan, specifically for training in this new subspecialty. During a period of 14 years, from 1986 to the present, an ad hoc committee comprising senior executive committee members of the American Society of Interventional and Therapeutic Neuroradiology, the Joint Section of Cerebrovascular Neurosurgery, and the American Society of Neuroradiology met to establish general guidelines that were mutually agreeable to both societies for training physicians in this field. These training standard guidelines were unanimously endorsed by the Executive Committee of the Joint Section of Cerebrovascular Neurosurgery in April 1999, by the Executive Committee of the American Society of Interventional and Therapeutic Neuroradiology and the American Society of Neuroradiology in May 1999, and by the Executive Council of the American Association of Neurological Surgeons and the Congress of Neurological Surgeons in June 1999. The guidelines for residency/ fellowship education have now been endorsed by the parent organizations of both the interventional and diagnostic neuroradiology communities, as well as both senior organizations representing neurosurgery in North America. These guidelines for training should be used as a reference and guide by any institution establishing a training program in neuroendovascular surgery/interventional neuroradiology. PMID- 10834653 TI - Neurogenic tumors of the cervical vagus nerve: report of four cases and review of the literature. AB - OBJECTIVE AND IMPORTANCE: Nerve sheath tumors arising from the cervical vagus nerve are extremely rare. These tumors most often present as asymptomatic, slowly enlarging, lateral neck masses and therefore often come initially to the attention of otolaryngologists and general surgeons. Because they are nerve tumors, however, neurosurgeons must be able to recognize and treat these rare entities. We report three cases of schwannoma and one case of neurofibroma of the cervical vagus nerve that were encountered at our center (Louisiana State University Medical Center) during a 31-year period. CLINICAL PRESENTATION: The patients ranged from 31 to 61 years of age at the time of presentation to Louisiana State University Medical Center. Presenting complaints included hoarseness, Horner's syndrome, and palpation of an enlarging, asymptomatic, cervical mass. Reviews of systems revealed episodes of aspiration for one patient and frequent respiratory illnesses for two patients. These episodes were possibly related to their tumors. Imaging studies demonstrated well-circumscribed masses in the region of the carotid sheath. INTERVENTION: Using microsurgical techniques, gross total resection of all four tumors was accomplished. For one patient, the vagus nerve needed to be divided and an end-to-end anastomosis was performed. For the other three patients, resection of the tumor was achieved with the vagus nerve in continuity. CONCLUSION: Vagal nerve schwannomas and neurofibromas in the neck are rare neoplasms. We present four cases of these benign tumors. The pathological features, epidemiological characteristics, presentation, differential diagnosis, and management are discussed. Gross total resection with preservation of the vagus nerve remains the treatment of choice. PMID- 10834654 TI - Primary angiitis of the central nervous system as a first presentation in Hodgkin's disease: a case report and review of the literature. AB - OBJECTIVE AND IMPORTANCE: Granulomatous angiitis of the central nervous system is a rare cause of neurological deterioration. It is often diagnosed posthumously, and a high index of suspicion is necessary to make the correct diagnosis on a timely basis. CLINICAL PRESENTATION: A 27-year-old woman presented to the emergency room with complaints of worsening headache, nausea, and vomiting for 10 days, which were preceded by swelling of her tongue. At the examination, she had mild ocular tenderness, but no cranial nerve abnormalities. Radiographic examination revealed a right temporal lobe area with edema, and mild contrast enhancement was noted on computed tomography and magnetic resonance imaging. A similar but smaller region was present in the left frontal lobe. INTERVENTION: Stereotactic biopsy of the left temporal lobe revealed granulomatous angiitis. Further workup revealed Hodgkin's disease in the mediastinum. Dexamethasone as well as chemotherapy for Hodgkin's disease was initiated. The patient's symptoms resolved, and she returned to work with her disease in remission. CONCLUSION: Previous reports of central nervous system angiitis have shown an association with Sjogren's syndrome, herpes zoster infection, human immunodeficiency virus, and Hodgkin's disease. A review of the literature revealed a total of 12 patients with central nervous system angiitis and Hodgkin's disease. As a group, these patients had very poor outcomes. However, of six patients who presented with central nervous system angiitis and concurrent Hodgkin's disease and who underwent aggressive treatment for both conditions, three had a full recovery, two had a partial recovery, and one died. PMID- 10834655 TI - Chiasmal apoplexy due to hemorrhage from a pituitary adenoma into the optic chiasm: case report. AB - OBJECTIVE AND IMPORTANCE: Chiasmal apoplexy, defined as hemorrhage into the optic chiasm, generally is caused by an intrachiasmal vascular malformation. We report the first case of chiasmal apoplexy due to hemorrhage from a pituitary macroadenoma into the optic chiasm. CLINICAL PRESENTATION: A 52-year-old man presented with headache, sudden and severe deterioration of visual acuity in the left eye, and a bitemporal visual field deficit. Magnetic resonance imaging revealed a large intra- and suprasellar homogeneously enhancing mass, which elevated a markedly thickened optic chiasm. After emergent transsphenoidal resection of the pituitary adenoma, vision did not improve. INTERVENTION: A pterional craniotomy was subsequently performed, during which a hematoma was found and evacuated from within the substance of the left optic nerve and chiasm. The hematoma cavity was found to communicate with the sella through a defect in the diaphragm. Vision improved dramatically after the operation. CONCLUSION: Chiasmal apoplexy resulting from pituitary adenoma should be distinguished from pituitary apoplexy, particularly because it requires a different surgical treatment. Clinical and radiographic features that may help distinguish the two are discussed. PMID- 10834656 TI - Rupture of a large ophthalmic segment saccular aneurysm associated with closed head injury: case report. AB - OBJECTIVE AND IMPORTANCE: Although each year approximately 30,000 to 50,000 cases of subarachnoid hemorrhage in the United States are caused by the rupture of intracranial saccular aneurysms, there is little information in the literature documenting the association of aneurysmal rupture with closed head injury. CLINICAL PRESENTATION: A 61-year-old woman presented after a motor vehicle accident with multiple injuries, including a severe closed head injury. Computed tomography revealed a diffuse basal subarachnoid hemorrhage. Angiography revealed the source as a large aneurysm arising from the ophthalmic segment of the left carotid artery. INTERVENTION: After the patient was stabilized for her multiple injuries, she underwent craniotomy and clipping of the aneurysm. She recovered without developing new neurological deficits. CONCLUSION: Although the association of head trauma and aneurysmal subarachnoid hemorrhage is rare, the presence of significant basal subarachnoid blood on a computed tomographic scan should alert the physician to the possibility of a ruptured aneurysm. PMID- 10834657 TI - Successful management of sellar and suprasellar arachnoid cysts with stereotactic intracavitary irradiation: an expanded report of four cases. AB - OBJECTIVE: Sellar and suprasellar arachnoid cysts may be asymptomatic or may cause headache, optic nerve compression, endocrine dysfunction, or hydrocephalus. We propose a minimally invasive treatment strategy when intervention is indicated. METHODS: Four patients with sellar and suprasellar arachnoid cysts presented with headache, visual compromise, and endocrine dysfunction. Two of the four patients previously had undergone unsuccessful surgical intervention. The imaging studies of two patients were diagnostic of an arachnoid cyst. RESULTS: All four patients underwent stereotactic intracavitary radiation with cyst regression and symptomatic improvement. In each patient, the optic chiasm was decompressed successfully. There were no complications from the procedure. CONCLUSION: Stereotactic intracavitary irradiation of arachnoid cysts proved to be safe and effective. The procedure obviated the need for open cyst fenestration or shunting. PMID- 10834658 TI - Transradial approach for vertebral artery stenting: technical case report. AB - OBJECTIVE: We report the technique of transradial vertebral artery stenting for two patients in whom severe supra-aortic ectasia prevented ready access to the right vertebral artery origin. METHODS: An Allen test was performed to verify ulnar artery collateral flow in the hand. After the administration of local anesthesia, a 6-French sheath was introduced into the radial artery. To allay induced spasm, a mixture of heparin (5000 IU/ml), verapamil (2.5 mg), nitroglycerine (400 microg/ml, 0.25 ml), and lidocaine (2%, 1.0 ml) was infused through the introducer sheath. A microguidewire was positioned across the vertebral artery lesion, followed by placement of a balloon-expandable stent. RESULTS: Postdeployment angiography revealed improved vertebrobasilar system flow. There were no complications related to the radial artery. The patients tolerated the procedure without difficulty and were immediately ambulatory. CONCLUSION: This novel approach should be considered for endovascular procedures for which access to the vertebral artery origin via the femoral approach is limited. PMID- 10834659 TI - Transcatheter snare removal of acute middle cerebral artery thromboembolism: technical case report. AB - OBJECTIVE AND IMPORTANCE: We describe the case of a patient in whom a snare designed for the removal of foreign bodies was successfully used to retrieve a thromboembolism from the middle cerebral artery. This technique can be used to reestablish blood flow when maximal pharmacological therapies have failed. CLINICAL PRESENTATION: A 38-year-old man with scrotal squamous cell carcinoma presented with the abrupt onset of left hemiparesis and numbness. Computed tomography of the head showed no hemorrhage or hypodensity, and right middle cerebral artery thrombosis was suspected. INTERVENTION: Cerebral angiography demonstrated a near-total occlusion of the right middle cerebral artery at the M1 M2 junction. The administration of intra-arterial urokinase, systemic heparin, and systemic abciximab, and mechanical maceration failed to lyse the clot. A 4-mm goose-neck snare was guided through a microcatheter, and the clot was snared and withdrawn. Immediate postoperative angiography demonstrated the reconstitution of normal flow. Pathological examination of the snared material was consistent with clot. By postoperative Day 5, the patient had regained full strength, except for the fingers of the left hand, which remained moderately weak. Computed tomography demonstrated a right insular and extreme capsular infarct. CONCLUSION: To our knowledge, this is the first reported use of a snare to remove clot in the setting of thromboembolic stroke. As the use of intra-arterial thrombolysis increases, transcatheter snare removal of pharmacologically resistant clot may be considered as a salvage strategy. PMID- 10834660 TI - New aneurysm clip and applier for narrow spaces: technical note. PMID- 10834661 TI - Endoscopic third ventriculostomy: outcome analysis of 100 consecutive procedures. PMID- 10834662 TI - Bilateral anterior cingulotomy for chronic noncancer pain. PMID- 10834663 TI - Relative contribution of preload and afterload to the reduction in cardiac output caused by nitric oxide synthase inhibition with L-N(G)-methylarginine hydrochloride 546C88. AB - OBJECTIVE: The nitric oxide synthase inhibitor L-N(G)-methylarginine hydrochloride (L-NMMA HC1 546C88) causes reductions in cardiac output (CO), a potential limitation to clinical application. This drop in CO exceeds that from phenylephrine at matched systemic arterial pressure. We tested the hypothesis that the greater fall in CO attributable to L-NMMA primarily reflects a difference in venoconstriction between agents, such that phenylephrine produces larger increases in preload (an independent determinant of CO). DESIGN: Random infusion of phenylephrine or L-NMMA. SETTING: An animal research laboratory. SUBJECTS: Eight healthy, conscious, male dogs. INTERVENTIONS: L-N(G) methylarginine hydrochloride (20 mg/kg for 1 hr) and phenylephrine (0.5 to 3 microg/kg/min) were administered into eight dogs chronically instrumented to measure left ventricular pressure and dimension. Data were measured at a constant heart rate (140 beats/min) to render CO proportional to stroke dimension. MEASUREMENTS AND MAIN RESULTS: At a matched increase in afterload (effective arterial elastance), L-NMMA increased preload (end-diastolic dimension) to a lesser degree (3.8%+/-1.5%, p < .05) than phenylephrine (9.6%+/-1.6%, p < .05 vs. L-NMMA). Neither L-NMMA nor phenylephrine affected the slope of the end-systolic pressure dimension relationship, although L-NMMA shifted the relationship rightward (1.7+/-0.7 mm, p < .05), consistent with a mild negative inotropic effect. L-NMMA decreased the stroke dimension to a greater extent than phenylephrine (-24.1%+/-6.8% and -10.6%+/-3.4%, respectively, p < .05). CONCLUSIONS: Differential CO responses to phenylephrine and L-NMMA were primarily attributable to changes in preload. Variable venular vs. arteriolar constrictor effects must be considered when evaluating the integrated cardiovascular response to a vasoactive agent. PMID- 10834664 TI - Pressure support ventilation in patients with acute lung injury. AB - OBJECTIVES: To assess the success rate of pressure support ventilation (PSV) in acute lung injury patients undergoing continuous positive pressure ventilation (CPPV), to study physiologic changes after the transition from CPPV to PSV, and to investigate differences between patients who succeed and patients who fail PSV according to predetermined criteria. DESIGN: Observational study. SETTING: General intensive care unit in a teaching hospital. SUBJECTS: We studied 48 patients having acute lung injury, as defined by a PaO2/F(IO2) <300 mm Hg and the presence of bilateral infiltrates on chest radiograph, and ventilated with CPPV. We included patients with PaO2 >80 mm Hg, at positive end-expiratory pressure of <15 cm H2O and with F(IO2) up to 1.0. INTERVENTIONS: After enrollment, PSV was instituted and patients were strictly monitored during the following 48 hrs. Subjects who met any of the predefined PSV failure criteria during this period were returned to CPPV (Group F). PSV was continued in the remaining patients (Group S). MEASUREMENTS AND MAIN RESULTS: Gas exchange, respiratory mechanics, and hemodynamics measurements were collected before switching from CPPV to PSV and were repeated at 24 hrs after beginning PSV, or immediately before return to CPPV in Group F patients. The physiologic deadspace volume to tidal volume ratio (V(D)/V(T)) was obtained by the Enghoff's equation from the measurement of the mixed expired CO2 fraction. PSV resulted in a significant PaCO2 decrease (49.2+/ 10.9 mm Hg to 44.4+/-7.2 mm Hg) and significant increases in minute volume (V(E))(9.0+/-2.3 L/min to 12.0+/-4.0 L/min) and arterial blood pH (7.405+/-0.054 to 7.435+/-0.064), with stable oxygenation and hemodynamics. In patients who were hypercapnic (PaCO2 >50 mm Hg) during CPPV, the V(E) increase was higher than in normocapnic patients. In the latter patients, PaCO2 and pH did not change significantly going from CPPV to PSV. A total of 38 patients (79%) were allocated to Group S and the remaining 10 patients were included in Group F. In Group S, positive endexpiratory pressure of 9.4+/-2.9 cm H2O (range, 3-14 cm H2O) and a PSV level of 14.9+/-3.8 cm H2O (range, 9-22 cm H2O) were applied. In Group F, positive end-expiratory pressure of 8.9+/-3.1 cm H2O (range, 5-15 cm H2O) and a PSV level of 21.6+/-4.6 cm H2O (range, 16-31 cm H2O) were adopted. Compared with Group S, Group F had a longer duration of intubation (20.2+/-19.2 days vs. 9.2+/ 13.5 days), a lower static compliance of the respiratory system (30.4+/-16.5 mL/cm H2O vs. 41.7+/-15.0 mL/cm H2O), and a higher V(D)/V(T) (0.70+/-0.09 vs. 0.52+/-0.10), but similar oxygenation and positive end-expiratory pressure. V(E) was higher in Group F during both CPPV and PSV. CONCLUSIONS: In a relatively high proportion of the investigated patients, PSV was successful. The institution of PSV led to no major changes in oxygenation or in hemodynamics. PSV was associated with increases in V(E) and respiratory frequency. In patients who had been hypercapnic during CPPV, PaCO2 decreased despite a compensated pH. Compared with PSV success patients, patients who failed PSV appeared to be sicker, as shown by the higher duration of respiratory support, increased ventilatory needs, and decreased respiratory system compliance, despite similar arterial oxygenation and positive end-expiratory pressure. PMID- 10834665 TI - Secretory leukocyte protease inhibitor, an inhibitor of neutrophil activation, is elevated in serum in human sepsis and experimental endotoxemia. AB - OBJECTIVES: To document changes in serum secretory leukocyte protease inhibitor (SLPI) in human sepsis and in experimental endotoxemia in vivo. To compare changes in serum SLPI in human sepsis with changes in interleukin (IL)-6, IL-10, and tumor necrosis factor (TNF)-alpha. To determine whether or not changes in SLPI correlate with the severity of multiple organ dysfunction syndrome as measured by the maximal multiple organ dysfunction score. Finally, because neutrophils have been implicated in tissue injury associated with organ dysfunction, to determine whether recombinant human SLPI blocks activation of isolated human neutrophils. DESIGN: Case-control study and ex-vivo cellular assay. SETTING: Surgical intensive care unit and clinical research center of university hospitals; laboratory of a medical school. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: There was a significant dose-dependent elevation (50.2+/-4.0 ng/mL, p = .01) in plasma SLPI 12 hrs after administration of lipopolysaccharide to seven healthy adults (36.4+/-2.3 ng/mL). Further, serum concentrations of SLPI (132+/-15 ng/mL) were elevated in septic surgical patients compared with healthy controls (43+/-2 ng/mL, p < .01) and nonseptic surgical controls (69+/-10 ng/mL, p = .01). Serum SLPI concentrations correlated (r2 = .71, p < .01) better with organ dysfunction as measured by maximal multiple organ dysfunction score than did serum IL-6 (r2 = .49, p < .01), IL-10 (r2 = .05, p = .22), or TNF-alpha (r2 = .02, p = .44). We found that recombinant human SLPI in vitro inhibits TNF-alpha-induced hydrogen peroxide production by human neutrophils (ID50 = 1-2 microg/mL). CONCLUSIONS: Serum SLPI is elevated in human sepsis and experimental endotoxemia. Maximal concentrations of serum SLPI correlate significantly with maximal multiple organ dysfunction scores in patients with sepsis. Secretory leukocyte protease inhibitor may function to limit ongoing neutrophil-mediated tissue injury associated with organ dysfunction. PMID- 10834666 TI - Effect of different recumbent positions on spectral indices of autonomic modulation of the heart during the acute phase of myocardial infarction. AB - OBJECTIVE: To examine which recumbent position can lead to the highest vagal modulation in patients during the acute phase of myocardial infarction. DESIGN: Descriptive study. SETTING: Intensive care unit in a medical center. PATIENTS: A total of 52 patients admitted to the intensive care unit because of acute myocardial infarction (AMI), 41 patients with coronary artery disease (CAD), and 28 patients with patent coronary arteriogram. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Heart rate variability analysis was performed in patients with AMI, patients with CAD, and patent coronary controls in supine, left lateral decubitus, and right lateral decubitus positions in random order. In the right lateral decubitus position, the vagal modulation was the highest and the sympathetic modulation was the lowest among three recumbent positions in three groups of patients. When the position was changed from supine to right lateral decubitus, the increase in vagal modulation was greater in patients who had more severely depressed vagal modulation in the supine position and the rate of increase was the greatest in patients with AMI, followed by patients with CAD and patent coronary controls. Detailed analysis showed that the vagal enhancing and sympathetic suppression effect of the right lateral decubitus position applied to patients with Q wave myocardial infarctions. CONCLUSIONS: The right lateral decubitus position can lead to the highest vagal modulation and the lowest sympathetic modulation among three recumbent positions in patients with Q wave myocardial infarction. The right lateral decubitus position can be used as an effective vagal enhancer in patients with Q wave myocardial infarction but without severe bradycardia or atrioventricular block. PMID- 10834667 TI - Microvascular response in patients with cardiogenic shock. AB - OBJECTIVE: To examine the mechanisms contributing to decreased microvascular blood flow in cardiogenic shock by comparing patients with cardiogenic shock with critically ill controls and with patients with septic shock. DESIGN: Prospective, consecutive entry of patients meeting the criteria for septic shock, cardiogenic shock, and critical illness without coexisting infection or shock. SETTING: University hospital, medical intensive care unit, coronary care unit, and respiratory care unit. PATIENTS: Eight patients with cardiogenic shock secondary to acute myocardial infarction, six critically ill controls, and six patients with septic shock. MEASUREMENTS AND MAIN RESULTS: Forearm blood flow was measured at rest and during reactive hyperemia by venous air plethysmography. Red cell deformability was determined by filtration. Leukocyte aggregation was detected by the leukergy test. Neutrophil CD11b/CD18 expression and soluble intercellular adhesion molecule-1 levels were also measured. In cardiogenic shock, forearm arterial resistance was significantly increased at rest and during reactive hyperemia compared with controls and patients with septic shock. The response to reactive hyperemia was attenuated in cardiogenic and septic shock patients, as measured by the absolute change in forearm blood flow from baseline, which was significantly less as compared with controls (p < .01). The percent change in forearm blood flow during reactive hyperemia compared with forearm blood flow at rest was significantly lower in cardiogenic shock (60+/-10) and in septic shock (50+/-11) compared with controls at baseline (145+/-20; p < .01). Red cell deformability was significantly decreased in cardiogenic shock (1.2+/-0.2 mL/min; p < .05) and septic shock (1.1+/-0.2 mL/min; p < .05), compared with controls (1.8+/-0.1 mL/min). Neutrophil CD11b/CD18 expression, leukergy, and serum intercellular adhesion molecule-1 levels in cardiogenic shock patients were not significantly different from controls. CONCLUSION: These data suggest that the response to reactive hyperemia is attenuated in cardiogenic shock. This appears to reflect increased vasoconstriction and an impaired capacity for vasodilation. Decreased erythrocyte deformability may also be important in limiting systemic microvascular flow. However, evidence supporting a role for neutrophil endothelial cell interactions was not observed. PMID- 10834668 TI - Alfentanil reduces the febrile response to interleukin-2 in humans. AB - OBJECTIVE: Manifestation of intraoperative fever is impaired by volatile anesthetics and muscle relaxants. Opioids are common anesthetic adjuvants and remain the dominant treatment for postoperative surgical pain and sedation of critically ill patients. The effect of opioids on normal thermoregulatory control is well established. However, the extent to which these drugs might inhibit fever remains unknown. Accordingly, we tested the hypothesis that relatively low plasma concentrations of the mu-receptor agonist alfentanil reduce fever magnitude. DESIGN: Prospective, randomized, crossover study. SETTING: Outcomes Research Laboratory, at the Department of Anesthesia and Perioperative Care, University of California, San Francisco. PATIENTS: Eight healthy male volunteers, aged 25-31 yrs, each studied on three separate days. INTERVENTION: Each volunteer was given an intravenous injection of 30 IU/g interleukin (IL)-2, followed 2 hrs later by 70 IU/g. One hour after the second dose, the volunteers were randomly assigned to three doses of alfentanil: a) none (control); b) a target plasma concentration of 100 ng/mL; and c) a target concentration of 200 ng/mL. Opioid administration continued for 5 hrs. METHODS AND MAIN RESULTS: Alfentanil significantly reduced the febrile response to pyrogen, decreasing integrated tympanic membrane temperatures from 7.5+/-2.2 degrees C x hr on the control day, to 4.9+/-1.5 degrees C x hr with 100 ng/mL alfentanil, and to 5.1+/-1.7 degrees C x hr with 200 ng/mL alfentanil (p = .011). Peak temperatures were also significantly reduced from 38.5+/-0.4 degrees C on the control day, to 38.0+/-0.4 degrees C on the 100 ng/mL-alfentanil day and 38.0+/-0.6 degrees C on the 200-ng/mL day (p = .019). Plasma cytokine concentrations increased after IL-2 administration, roughly in proportion to the elevation in core temperature. However, cytokine concentrations did not differ significantly among the treatment groups. CONCLUSION: Alfentanil significantly reduced the febrile response to IL-2 administration. However, the reduction was comparable at plasma concentrations near 100 and 200 ng/mL. These data indicate that concentrations of opioids commonly observed in critical care patients significantly inhibit the manifestation of fever. PMID- 10834669 TI - Predictors of mortality in stroke patients admitted to an intensive care unit. AB - OBJECTIVE: Improved pathophysiologic insight and prognostic information regarding in-hospital risk of mortality among stroke patients admitted to an intensive care unit. DESIGN: Retrospective analysis. SETTING: Neurology/neurosurgery intensive care unit in a tertiary care university medical center. PATIENTS: A total of 63 consecutive ischemic stroke patients. INTERVENTIONS: Patients were classified according to in-hospital mortality. Charts were reviewed to retrospectively generate an admitting Acute Physiology and Chronic Health Evaluation (APACHE) II score. The APACHE II score and its individual components were assessed for predicting subsequent death. MEASUREMENTS AND MAIN RESULTS: Of 63 patients, 13 died and 50 survived to either discharge or surgical intervention. The mean admitting APACHE II score of survivors (6.9) was lower than that of patients who died (17.2; p < .0001). None of the 33 patients with a score <9 died, compared with 43% of those with a score > or =9. A score > or =18 was uniformly associated with fatal outcome (n = 8). Univariate analysis identified APACHE II total score, Glasgow Coma Scale score, temperature, pH, and white blood cell count as significant predictors of death. Among multivariate logistic regression models examining the components of the APACHE II score, the model containing white blood cells, temperature, and creatinine best predicted death. CONCLUSIONS: Several features of the APACHE II score are associated with risk of death in this patient population. The findings suggest particular physiologic derangements that are associated with, and may contribute to, increased mortality in critically ill patients with acute ischemic stroke. PMID- 10834670 TI - Head computed tomography in medical intensive care unit patients: clinical indications. AB - OBJECTIVE: To assess whether clinical variables might be useful in selecting patients who will have an acute intracranial abnormality seen in head computed tomographic scans (HCT). DESIGN: Retrospective study. SETTING: Medical intensive care unit (MICU) in a tertiary teaching hospital. MEASUREMENTS: Medical records of patients admitted to the MICU who underwent HCT between January 1, 1994, and December 31, 1995, were reviewed. Patients with acute intracranial abnormalities (HCT-positive) and those without new acute findings (HCT-negative) were compared on various clinical variables, including demographics, indications for obtaining the HCT (mental status change, neurologic deficit, fever, seizures), coagulation profiles, when the HCT was performed (at admission or after admission), and ordering physician. MAIN RESULTS: Of 297 HCTs obtained in 230 patients, 37% (109/297) were positive. When the clinical variables were examined univariately, only the presence of a neurologic deficit (70% vs. 37%; difference, 33%; p < .001) differed significantly between positive and negative HCTs. Multivariate analysis confirmed that only the frequency of a new neurologic deficit differed significantly in the two groups (p < .001; odds ratio, 3.9; 95% confidence interval, 2.3-6.4). In patients without neurologic deficits, only the presence of seizures was associated with a positive HCT (p < .01: logistic regression). The presence of either neurologic deficit or seizures best predicted a positive HCT: sensitivity 0.81, specificity 0.53, positive predictive value 0.50, and negative predictive value 0.83. CONCLUSION: Among MICU patients, the presence of either neurologic deficit or seizures is associated with the presence of an acute intracranial abnormality seen in HCT, but the association is not powerful enough to reliably depend on these clinical variables to select patients for HCTs in the MICU. PMID- 10834671 TI - Metabolic encephalopathy in critically ill patients suffering from septic or nonseptic multiple organ failure. AB - OBJECTIVE: Evaluation of changes in the peak latencies of sensory evoked potentials in different patient groups, to evaluate differences in metabolic encephalopathy of critically ill patients with multiple organ failure as a result of septic or nonseptic conditions. DESIGN: Prospective cohort study. SETTING: Intensive care units of the university hospital, Vienna. PATIENTS: Patients (n = 103) treated on an intensive care unit because of multiple organ failure with additional metabolic encephalopathy. Multiple organ failure was induced by sepsis (group A; n = 56), surgery (group B; n = 29), or both (group C; n = 18). INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Metabolic encephalopathy was determined by measuring median nerve-stimulated short-latency and long-latency sensory evoked potentials. No differences in the peak latencies of the sensory evoked potentials were detected among the groups. Septic patients had a N70 peak latency of 131+/-21 msecs, nonseptic postsurgical patients of 132+/-17 msecs, and septic postsurgical patients of 134+/-17 msecs. The cervicomedullary N13 to cortical N20 conduction times were 6.4+/-1 msec, 6.4+/-1.4 msecs, and 6.8+/-1.2 msecs, respectively. All measured peak latencies were significantly prolonged compared with peak latencies of healthy controls. The severity of illness assessed by the Acute Physiology and Chronic Health Evaluation III score was not different between the three groups. An increase of the delay of N70 peak latencies was significantly correlated with the severity of illness (r2 = .15; p < .00005). CONCLUSION: There was no difference in sensory evoked potential measurements detectable among septic patients with multiple organ failure, nonseptic postsurgical patients with multiple organ failure, and septic postsurgical patients with multiple organ failure. The N70 peak latency was significantly correlated with the severity of illness but not with the presence or absence of sepsis. In postsurgical patients with multiple organ failure and superimposed sepsis, the N70 peak latencies were not further prolonged compared with postsurgical patients without sepsis. PMID- 10834672 TI - Prevalence of infections in intensive care units in Mexico: a multicenter study. AB - OBJECTIVE: To determine the 1-day prevalence of community-acquired, hospital acquired, or intensive care unit (ICU)-acquired infections in Mexican ICUs. To identify associated risk factors, predominant infecting organisms, and mortality rates. DESIGN: A 1-day point-prevalence study. SETTING: A total of 254 adult ICUs in Mexico. PATIENTS: Adult patients hospitalized in the participating ICUs. RESULTS: A total of 895 patients were studied, of whom 521 patients (58.2%) were infected. Community-acquired infection occurred in 214 patients (23.9%), non-ICU nosocomial infection occurred in 99 patients (11.1%), and 208 patients had at least one ICU-acquired infection (23.2%; 1.45 episodes/patient). The most frequently reported ICU-acquired infections were pneumonia (39.7%), urinary tract infections (20.5%), wound infection (13.3%), and bacteremia (7.3%). The mortality rate for the ICU-acquired infections after 6 wks of follow-up was 25.5%. Multivariate regression analysis showed the following risk factors for ICU acquired infections: neurologic failure as a primary cause of admission (odds ratio [OR], 1.697; 95% confidence interval [CI], 1.001-2.839); length of stay in ICU (OR, 1.119; 95% CI, 1.091-1.151); number of therapeutic and/or diagnostic interventions during the preceding week (OR, 1.118; 95% CI, 1.016-1.231); peripherally administered infusion of hyperosmolar solutions (OR, 6.93; 95% CI, 2.452-21.661); sedative usage in the preceding week (OR, 1.751; 95% CI, 1.183 2.602); history of an emergency surgery in the preceding month (OR, 1.875; 95% CI, 1.251-2.813). The administration of antimicrobial treatment if there was an infection decreased the risk of death (OR, 0.406; 95% CI, 0.204-0.755). CONCLUSIONS: Evidence of a high frequency of nosocomial infections was found, and potential risk factors for acquiring infections and mortality were identified. Mortality rates according to the hierarchy of the systemic inflammatory response syndrome in Latin American ICUs are reported. PMID- 10834673 TI - Outcome and prognostic factors in critically ill cancer patients admitted to the intensive care unit. AB - OBJECTIVE: To assess survival in cancer patients admitted to an intensive care unit (ICU) with respect to the nature of malignancy, cause of ICU admittance, and course during ICU stay as well as to evaluate the prognostic value of the Acute Physiology and Chronic Health Evaluation (APACHE) III score. DESIGN: Retrospective cohort study. SETTING: ICU at a university cancer referral center. PATIENTS: A total of 414 cancer patients admitted to the ICU during a period of 66 months. INTERVENTIONS: None. MEASUREMENTS: Charts of the patients were analyzed with respect to underlying disease, cause of admission, APACHE III score, need and duration of mechanical ventilation, neutropenia and development of septic shock, as well as ICU survival and survival after discharge. Mortality data were compared with two control groups: 1362 patients admitted to our ICU suffering from diseases other than cancer and 2,776 cancer patients not admitted to the ICU. MAIN RESULTS: ICU survival was 53%, and 1-yr survival was 23%. The 1 yr mortality rate was significantly lower in both control groups. Patients admitted after bone marrow transplantation had the highest mortality. In a multivariate analysis, prognosis was negatively influenced by respiratory insufficiency, the need of mechanical ventilation, and development of septic shock during the ICU stay. Admission after cardiopulmonary resuscitation yielded high ICU mortality but a relatively good long-term prognosis. Admission after surgery and as a result of acute hemorrhage was associated with a good prognosis. Age, neutropenia, and underlying disease did not influence outcome significantly. Admission APACHE III scores were significantly higher in nonsurvivors but failed to predict individual outcome satisfactorily. All patients with APACHE III scores of >80 died at the ICU. CONCLUSION: A combination of factors must be taken into account to estimate a critically ill cancer patient's prognosis in the ICU. The APACHE III scoring system alone should not be used to make decisions about therapy prolongation. Admission to the ICU worsens the prognosis of a cancer patient substantially; however, as ICU mortality is 47%, comparable with severely ill noncancer patients, general reluctance to admit cancer patients to an ICU does not seem to be justified. PMID- 10834674 TI - Prolonged low-dose dopamine infusion induces a transient improvement in renal function in hemodynamically stable, critically ill patients: a single-blind, prospective, controlled study. AB - OBJECTIVE: To evaluate the length of the effects of long-term (48 hrs), low-dose dopamine infusion on both renal function and systemic hemodynamic variables in stable nonoliguric critically ill patients. DESIGN: Prospective, single-blind, controlled clinical study. SETTING: University hospital, 19-bed multidisciplinary intensive care unit. PATIENTS: Eight hemodynamically stable, critically ill patients with a mild nonoliguric renal impairment (creatinine clearance between 30 and 80 mL/min). INTERVENTIONS: Each patient consecutively received 4 hrs of placebo, followed by a 3 microg/kg/min dopamine infusion during 48 hrs, then a new 4-hr placebo period. We measured cardiac output and other hemodynamic variables by using a pulmonary artery catheter. The bladder was emptied to determine urine volume and to collect urine samples. Measurements were performed at six times: after the initial control of 4 hrs of placebo (C1); after 4 hrs (H4), 8 hrs (H8), 24 hrs (H24), and 48 hrs (H48) of dopamine infusion; and after the second control of 4 hrs of placebo (C2). MEASUREMENTS AND MAIN RESULTS: We saw no significant change in systemic hemodynamic variables with dopamine at all times of infusion. Diuresis, creatinine clearance, and the fractional excretion of sodium (FENa) at C1 and C2 were not different. Urine flow, creatinine clearance, and FENa increased significantly 4 hrs after starting dopamine (for all these changes, p < .01 vs. C1 and C2). The maximum changes were obtained at H8, with an increase of 50% for diuresis, 37% for creatinine clearance, and 85% for FENa (for all these changes, p < .01 vs. C1 and C2). But these effects waned progressively from H24, and both creatinine clearance and FENa at H48 did not differ from control values. CONCLUSIONS: In stable critically ill patients, preventive low-dose dopamine increased creatinine clearance, diuresis, and the fractional excretion of sodium without concomitant hemodynamic change. These effects reached a maximum during 8 hrs of dopamine infusion. But despite a slight persistent increase in diuresis, improvement in creatinine clearance and FENa disappeared after 48 hrs. According to these data, it is likely that tolerance develops to dopamine-receptor agonists in critically ill patients at risk of developing acute renal failure. PMID- 10834675 TI - Effects of ventilation and nonventilation on pulmonary venous blood gases and markers of lung hypoxia in humans undergoing total cardiopulmonary bypass. AB - OBJECTIVE: To assess the effects of lung oxygenation and ventilation vs. lung collapse on pulmonary markers of lung hypoxia. DESIGN: A prospective, nonrandomized, nonblinded comparative study. SETTING: University department of anesthesiology and cardiothoracic surgery. SUBJECTS: Twelve adult patients undergoing coronary bypass grafting requiring total cardiopulmonary bypass. INTERVENTIONS: Single lung ventilation during total cardiopulmonary bypass (tidal volume, 150 mL; respiratory rate, 6 breaths/min; inspiratory oxygen fraction, 0.5) while the contralateral lung was allowed to collapse completely without oxygenation. MEASUREMENTS AND MAIN RESULTS: At the beginning and at the end of total cardiopulmonary bypass (duration, 59-65 mins), blood was aspirated from the right and left pulmonary veins and the radial artery for measurement of blood gases and concentrations of endothelin-1, big-endothelin, thromboxane B2, lactate, and lactate dehydrogenase. Nonventilation during total cardiopulmonary bypass compared with ventilation resulted in lower pulmonary venous P(O2) values (57+/-15 torr [7.6+/-2.0 kPa] vs. 103+/-23 torr [13.7+/-3.1 kPa]) and higher thromboxane B2 concentrations (488+/-95 pg/mL vs. 434+/-92 pg/mL). The concentrations of endothelin-1, big-endothelin, lactate, and lactate dehydrogenase in the pulmonary veins did not differ significantly between nonventilated and ventilated lungs. CONCLUSIONS: Development of pulmonary tissue hypoxia during 1 hr of nonventilation and cardiopulmonary bypass with completely inhibited pulmonary arterial blood flow is unlikely, suggesting that enough oxygen is stored in or is provided to the collapsed lung. Thus, nonventilation during total cardiopulmonary bypass does not appear to contribute to postoperative respiratory dysfunction by causing pulmonary tissue hypoxia. These results, however, do not exclude that mechanical factors of ventilation might benefit the lung during cardiopulmonary bypass. PMID- 10834676 TI - Prediction of post-extubation work of breathing. AB - OBJECTIVE: To evaluate which mode of preextubation ventilatory support most closely approximates the work of breathing performed by spontaneously breathing patients after extubation. DESIGN: Prospective observational design. SETTING: Medical, surgical, and coronary intensive care units in a university hospital. PATIENTS: A total of 22 intubated subjects were recruited when weaned and ready for extubation. INTERVENTIONS: Subjects were ventilated with continuous positive airway pressure at 5 cm H2O, spontaneous ventilation through an endotracheal tube (T piece), and pressure support ventilation at 5 cm H2O in randomized order for 15 mins each. At the end of each interval, we measured pulmonary mechanics including work of breathing reported as work per liter of ventilation, respiratory rate, tidal volume, negative change in esophageal pressure, pressure time product, and the airway occlusion pressure 100 msec after the onset of inspiratory flow, by using a microprocessor-based monitor. Subsequently, subjects were extubated, and measurements of pulmonary mechanics were repeated 15 and 60 mins after extubation. MEASUREMENTS AND MAIN RESULTS: There were no statistical differences between work per liter of ventilation measured during continuous positive airway pressure, T piece, or pressure support ventilation (1.17+/-0.67 joule/L, 1.11+/-0.57 joule/L, and 0.97+/-0.57 joule/L, respectively). However, work per liter of ventilation during all three preextubation modes was significantly lower than work measured 15 and 60 mins after extubation (p < .05). Tidal volume during pressure support ventilation and continuous positive airway pressure (0.46+/-0.11 L and 0.44+/-0.11 L, respectively) were significantly greater than tidal volume during both T-piece breathing and spontaneous breathing 15 mins after extubation (p < .05). Negative change in esophageal pressure, the airway occlusion pressure 100 msec after the onset of inspiratory flow, and pressure time product were significantly higher after extubation than during any of the three preextubation modes (p < .05). CONCLUSIONS: Work per liter of ventilation, negative change in esophageal pressure, the airway occlusion pressure 100 msec after the onset of inspiratory flow, and pressure time product all significantly increase postextubation. Tidal volume during continuous positive airway pressure or pressure support ventilation overestimates postextubation tidal volume. PMID- 10834677 TI - How well do surrogates assess the pain of seriously ill patients? AB - OBJECTIVE: To compare pain assessments made by seriously ill hospitalized patients and their surrogates and to understand factors that might lead to reporting differences. DESIGN: Cross-sectional study. SETTING: Five U.S. teaching hospitals. PATIENTS: Patients were 2,645 of 9,105 patients in the Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatments for whom there were both patient and surrogate interviews about pain. The majority of patients had acute respiratory failure, multiple organ system failure with sepsis, exacerbation of chronic obstructive pulmonary disease, and congestive heart failure. INTERVENTIONS: None. MEASUREMENTS: We queried patients and surrogates by using Likert-type scale questions about the frequency and severity of patients' pain. MAIN RESULTS: Overall, surrogates correctly estimated presence or absence of patients' pain 73.5% of the time (kappa = 0.47, 95% confidence interval [CI], 0.44, 0.50), overestimating 16.8% of the time and underestimating 9.7% of the time. Exact estimation of the presence and level of pain was considerably poorer, with only 53.0% of surrogates correctly assessing exact level of pain (kappa = 0.31; 95% CI, 0.28, 0.33; weighted kappa = 0.43; 95% CI, 0.41, 0.46). Patients' reported level of pain, days in hospital before study admission, time interval between patient and surrogate assessment, and study hospital were associated with inaccuracy in surrogates' estimation of patients' pain. CONCLUSIONS: Family members correctly assess the level of pain of hospitalized patients with moderate success. Further studies are needed to assess the accuracy of assessment of pain by family members compared with that of health professionals, and to determine whether surrogates' accuracy in estimating patients' pain can be improved. PMID- 10834678 TI - Effects of dopamine and epinephrine infusions on renal hemodynamics in severe malaria and severe sepsis. AB - OBJECTIVE: To describe and compare the effects of dopamine and epinephrine in various doses on renal hemodynamics and oxygen transport in patients with severe malaria and severe sepsis. DESIGN: Prospective, controlled, crossover trial. SETTING: The intensive care unit of an infectious diseases hospital in Viet Nam. PATIENTS: Fourteen patients with severe falciparum malaria and five with severe sepsis. INTERVENTIONS: In an open, crossover design, we observed the effects on renal and systemic hemodynamics and oxygen transport of separate stepped infusions of epinephrine and dopamine. We measured renal blood flow (RBF) and cardiac output by the thermodilution method using fluoroscopically guided catheters. Creatinine clearance at each time point was calculated from the renal plasma flow and the renal arteriovenous difference in plasma creatinine. MEASUREMENTS AND MAIN RESULTS: Dopamine at a "renal" dose (2.5 microg/kg/min) was associated with a mean (95% confidence interval) fractional increase in the absolute renal blood flow index (RBFI) of 37% (13% to 61%) and in RBF as a fraction of cardiac output (RBF/CO) of 35% (10% to 59%; p = .007 and p = .014, respectively). The consequent 39% (14% to 64%) increase in renal oxygen supply (p = .002) was accompanied by a 32% (20% to 44%) decrease in the renal oxygen extraction ratio (p = .0003), leading to no net change in renal oxygen consumption. At higher doses (10 microg/kg/min), both RBF and RBF/CO were not significantly different from baseline values and decreased further as the dose was reduced again. There was no obvious explanation for this hysteresis. There was no change in renal oxygen consumption throughout the study. Because lactic acidosis developed, epinephrine was only given to eight of the 19 patients, and the full stepped epinephrine infusion was given to four patients. Epinephrine infusion was associated, both in absolute terms and when compared with dopamine, with a significant increase in renal vascular resistance (p = .0008 and .0005, respectively), a decrease in RBF/CO (p = .002 and .03), and a compensatory increase in the renal oxygen extraction ratio (p = .005 and .0001). RBFI and renal oxygen consumption remained constant throughout the epinephrine infusion profile. Neither epinephrine nor dopamine significantly affected creatinine clearance or urine output. Twelve patients (63%) were in established renal failure (plasma creatinine, >3 mg/dL) at the time of the study, although the presence or absence of renal failure did not significantly influence the effects of the study drugs. However, overall, the presence of renal failure was associated with a lower mean renal oxygen consumption, a lower mean renal oxygen consumption as a fraction of systemic oxygen consumption, and a higher mean renal vascular resistance. CONCLUSION: Although dopamine increased and epinephrine decreased fractional renal blood flow, there was no evidence that either drug produced either a beneficial or a deleterious effect on renal oxygen metabolism or function at any of the doses investigated. PMID- 10834679 TI - Comparison of methods of measurements of oxygen consumption in mechanically ventilated patients with multiple trauma: the Fick method versus indirect calorimetry. AB - OBJECTIVE: The purpose of this study was to compare the measurements of whole body oxygen consumption determined by the Fick method and by indirect calorimetry in mechanically ventilated patients with multiple trauma. DESIGN: A prospective, correlational, within-subjects design. SETTING: Surgical intensive care unit of a Level I trauma center. PATIENTS: Thirty-eight mechanically ventilated adults with multiple injuries who received a pulmonary artery catheter within 24 hrs of admission to the surgical intensive care unit. MEASUREMENTS AND MAIN RESULTS: After the initial resuscitation, simultaneous measurements of oxygen consumption (V(O2) by the reverse Fick equation and by indirect calorimetry were performed every 6 hrs for 24 hrs in normothermic patients who were at rest for at least 30 mins. At each measurement period, the mean V(O2) values determined by indirect calorimetry were significantly greater than the mean V(O2) values determined by the Fick method (time 1: 172+/-38 vs. 125+/-47 mL/min/m2, p < .0001; time 2: 170+/-31 vs. 130+/-48 mL/min/m2, p < .0001; time 3: 170+/-32 vs. 132+/-53 mL/min/m2, p < .0001; time 4: 169+/-29 vs. 130+/-60 mL/min/m2, p < .0002). By using the Bland and Altman technique, the mean bias was 41+/-3.95 mL/min/m2. Correlation coefficients of VO2 values between methods of measurements were statistically significant (r2 = .32, p = .0001; r2 = .32, p = .0001; r2 = .33, p = .0001; r2 = .18, p = .0001). CONCLUSIONS: Indirect calorimetry should be the preferred standard for measurement of oxygen consumption in severely injured patients. PMID- 10834680 TI - Evaluation of the utility of computed tomography in the initial assessment of the critical care patient with chest trauma. AB - OBJECTIVE: To determine the utility of thoracic computed tomography (TCT) in the initial assessment of critically ill patients with chest injuries. DESIGN: Prospective observational study of cohorts. SETTING: Trauma intensive care unit (ICU) of a Spanish Level III hospital (US equivalent Level I). PATIENTS: Three hundred seventy-five patients with chest injuries were studied, grouped into two cohorts according to whether they underwent admission TCT (exposed cohort, group I, n = 104) or not (unexposed cohort, group II, n = 271). INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Demographic data, initial severity scores, and chest radiograph (CXR)-based diagnosis were collected in all patients as independent variables. In patients of group I, we also recorded the TCT-based diagnosis and any incidents, complications, or therapy changes resulting from the TCT. The need for and duration of mechanical ventilation, length of ICU stay, and ICU mortality were gathered in the whole sample as dependent variables. The admission data were similar in the two groups, except for a higher Injury Severity Score (ISS) and thoracic ISS in group I. TCT proved to be more sensitive than CXR in detecting pulmonary contusion, hemothorax, pneumothorax, and vertebral fractures and in identifying the faulty placement of chest drainage tubes. TCT findings induced therapy changes in approximately 30% of patients in group I. In the other dependent variables studied, there were no differences between the two groups. In the multivariate analysis, the TCT screening had no effects on the time on mechanical ventilation, length of ICU stay, or mortality. CONCLUSIONS: TCT detects more chest injuries in trauma patients than does CXR and induces therapy changes in a considerable number of patients. However, this does not translate into an improvement in clinical outcomes. PMID- 10834681 TI - The safety of prolonging the use of central venous catheters: a prospective analysis of the effects of using antiseptic-bonded catheters with daily site care. AB - OBJECTIVE: To determine rates of catheter colonization and catheter-related bloodstream infection (CRBSI) when antiseptic-bonded central venous catheters (CVCs) and standardized daily site care are used with no predetermined interval for removal. DESIGN: Prospective observational study. SETTING: Two major trauma centers. PATIENTS: All trauma patients admitted to two major trauma centers that received a CVC from May 1996 through May 1998. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Catheters were semiquantitatively cultured to identify bacterial colonization and CRBSI. Monitored variables included total catheter days, anatomical site of catheter insertion, and area in hospital of catheter insertion. CVC tips and intracutaneous segments were semiquantitatively cultured. A total of 460 (92%) of 501 catheters placed in 324 trauma patients were evaluable, representing 95.5% of all catheter days during the study period. Rates of catheter colonization and CRBSI were 5% (5/1000 catheter days) and 1.5% (1.511000 catheter days), respectively. Subclavian catheters were in place longer than femoral or internal jugular catheters (p < .0001), but the colonization rate was significantly lower (p = .03; relative risk, 0.34; 95% confidence interval, 0.15-0.77). No differences in CRBSI rates among anatomical sites or between catheters used < or =14 days and those used >14 days were identified. CONCLUSION: Femoral and internal jugular antiseptic-bonded CVCs develop bacterial colonization earlier than subclavian CVCs. Subclavian antiseptic-bonded CVCs combined with standardized daily site care may be safely used >14 days in trauma patients. PMID- 10834682 TI - Prediction and timing of tracheostomy in patients with infratentorial lesions requiring mechanical ventilatory support. AB - OBJECTIVES: To determine the frequency and predictors of successful extubations and tracheostomy in patients with infratentorial lesions requiring mechanical ventilation and to determine the optimal time for tracheostomy based on probability of successful extubation and in-hospital survival according to the duration of translaryngeal intubation. DESIGN: Retrospective chart review. SETTINGS: A neurocritical care unit at a university hospital. PATIENTS: A total of 69 patients with infratentorial lesions who were mechanically ventilated during their intensive care unit stay. MEASUREMENTS AND MAIN RESULTS: Of the 69 patients who were mechanically ventilated, 23 (33%) were successfully extubated. In logistic regression analysis, both the presence of a Glasgow Coma Scale score >7 at time of intubation (odds ratio, 4.8; 95% confidence interval, 1.2-21.7) and the absence of brainstem deficits (odds ratio, 4.3; 95% confidence interval, 1.3 16.7), were independently associated with successful extubation. After extubation, 11 patients were reintubated; seven were reintubated within the same day because of poor control over secretions, airway spasm, or hypoventilation. Tracheostomy was performed in 23 (33%) patients, of whom 19 were successfully weaned off mechanical ventilatory support over a mean period of 3.7+/-4.0 days after tracheostomy. Patients undergoing tracheostomy had a significantly longer intensive care unit stay (19.1+/-9.0 vs. 8.7+/-6.6 days, p < .01) and total hospital stay (34.8+/-18.7 vs. 20.1+/-9.9 days, p < .01) compared with patients who were successfully extubated. The probability of successful extubation or death before extubation or tracheostomy was 67% on the day of intubation, which decreased to 5.8% after translaryngeal intubation for >8 days. CONCLUSIONS: An aggressive policy toward tracheostomy is justified based on the low frequency of successful extubations and high frequency of extubation failures and tracheostomies in patients with infratentorial lesions. The decision regarding tracheostomy should be made on day 8 of mechanical ventilatory support because of the low probability of subsequent extubation or in-hospital death. PMID- 10834683 TI - The effect of sodium citrate in arterial catheters on acid-base and electrolyte measurements. AB - OBJECTIVE: To compare the effects of heparin or sodium citrate used to anticoagulate indwelling arterial catheters on acid-base and electrolyte measurements. DESIGN: Randomized controlled trial. SETTING: Medical-surgical university-affiliated intensive care unit. SUBJECTS: Twenty patients with indwelling arterial catheters. INTERVENTIONS: Patients were randomly allocated to have ten 1-mL aliquots of blood sampled serially from an arterial catheter maintained with either heparin or sodium citrate. A sample then obtained by arterial puncture provided true measurement values. Acid-base and electrolyte measurements of whole blood were obtained from each sample by means of a Coming 860 analyzer. MEASUREMENTS AND MAIN RESULTS: Contamination with sodium citrate lowered ionized calcium and pH but increased glucose and Pco2. Heparin produced negligible effects on those measurements. When sodium citrate was used, reliable measurements were not obtained for ionized calcium, pH, and glucose, even after 9 mL of blood had been discarded. However, reliable P(CO2) measurements were obtained after 2 mL of blood was discarded. CONCLUSIONS: Sodium citrate used to maintain arterial catheters can contaminate blood samples. The result of that contamination can mimic severe hypocalcemia, metabolic acidosis, and mild hyperglycemia. Failure to recognize the effects of sodium citrate on acid-base and electrolyte measurements may lead to changes in treatment that could affect patient outcome adversely. PMID- 10834684 TI - Exocrine pancreatic function in critically ill patients: septic shock versus non septic patients. AB - OBJECTIVE: To determine the exocrine pancreatic function in critically ill patients. DESIGN: Prospective cohort study. SETTING: Medical intensive care unit. PATIENTS: A total of 18 critically ill patients (11 patients with septic shock according to the criteria of the American College of Chest Physicians/Society of Critical Care Medicine, 7 nonseptic patients). The results obtained in these subjects were compared with the data obtained in seven healthy volunteers. INTERVENTIONS: Examination of exocrine pancreatic function was done by means of a cholecystokinin-secretin test. Intravenous stimulation of the exocrine pancreas with cholecystokinin and secretin (1 unit/kg body weight/hr each) and aspiration of duodenal fluid by a gastroscopically inserted oroduodenal tube was done during a period of 1 hr. MEASUREMENTS AND MAIN RESULTS: The content of amylase, chymotrypsin, and trypsin in aspirated duodenal fluid was significantly reduced in patients with septic shock compared with nonseptic patients as well as healthy subjects (p < .01). The volume of aspirated fluid was significantly reduced in patients with septic shock compared with healthy controls (p = .03), but not in nonseptic patients. The content of bicarbonate was not statistically different in the three groups. No significant correlation was to find between variables of exocrine pancreatic function and Acute Physiology and Chronic Health Evaluation III score, sepsis-related organ failure assessment score, systolic arterial pressure and mean arterial pressure in septic shock patients. Positive end expiratory pressure was significantly correlated with the content of trypsin (r2 = 0.52; p = .02). Postmortem examinations of five septic patients who died during the intensive care stay did not reveal gross morphologic alterations of pancreatic tissue. CONCLUSIONS: The study shows two pancreatic enzyme systems, namely, amylase as a carbohydrate splitting enzyme and the proteolytic enzymes trypsin and chymotrypsin, strongly affected in critically ill patients with septic shock. PMID- 10834685 TI - Percutaneous dilational tracheostomy or conventional surgical tracheostomy? AB - OBJECTIVE: Percutaneous dilational tracheostomy (PDT) is increasingly used in intensive care units (ICU), and it has a low incidence of complications. The aim of this study was to compare the costs, complications, and time consumption of PDT with that of conventional surgical tracheostomy (ST) when both procedures were performed in the ICU. DESIGN: The study was a prospective, randomized trial. SETTING: The procedures were performed routinely in the ICU of Satakunta Central Hospital. PATIENTS: During a 23-month period from December 1995 to November 1997, 30 patients underwent PDT and 26 patients had ST. In one patient, PDT was converted to ST. All patients were receiving ventilation in the ICU, and all tracheostomies were performed at the patient's bedside in the ICU. The Portex percutaneous tracheostomy kit was used for all PDTs. RESULTS: The mean time to perform PDT was 11 mins (SD, 6; range, 2-40), and the mean time to perform ST was 14 mins (SD, 6; range, 3-39). In the PDT group, five patients had moderate bleeding during the procedure. In three patients, the bleeding was resolved with compression; in one patient, it was resolved with ligation of the vessel; and in one patient, it was resolved with electrocoagulation. Bleeding did not cause any complications afterward. In the PDT group, one patient had minimal oozing from the wound edge on the first postoperative day and it was resolved spontaneously. In the ST group, there were no intraprocedural complications. One patient had bleeding from the wound on first postoperative day. The sutures were removed, and the bleeding vessel was ligated. The mean cost (in U. S. dollars) of PDT was $161 (SD, 10.4; range, $159-$219), and the mean cost of ST was $357 (SD, $74; range, $239-$599). The cost of PDT was significantly lower than the cost of ST (p < .001). CONCLUSION: We found that PDT is a cost-effective procedure in critically ill ICU patients. Although we performed ST at the bedside in the ICU to avoid the risks associated with moving critically ill patients to the operating room, we found PDT to be a simple and safe procedure. PMID- 10834686 TI - Plasma concentrations and anti-L-cytokine effects of alpha-melanocyte stimulating hormone in septic patients. AB - OBJECTIVES: The aim of this research was to investigate endogenous concentrations and anti-cytokine effects of the antiinflammatory peptide alpha-melanocyte stimulating hormone (alpha-MSH) in patients with systemic inflammation. The objectives were to determine the following: changes over time of plasma alpha-MSH and relationship with patient outcome, correlation between plasma alpha-MSH and tumor necrosis factor (TNF)-alpha plasma concentration and production in whole blood samples, and influences of alpha-MSH on production of TNF-alpha and interleukin (IL)-1beta in whole blood samples stimulated with lipopolysaccharide (LPS). DESIGN: Prospective, nonrandomized, clinical study. SETTING: Intensive care unit of a university hospital. PATIENTS: A total of 21 patients with sepsis syndrome/septic shock and an equal number of healthy volunteers. INTERVENTIONS: Circulating alpha-MSH and TNF-alpha concentrations and TNF-alpha production in supernatants of LPS (1 ng/mL)-stimulated whole blood were measured repeatedly. To determine whether alpha-MSH can modulate production of TNF-alpha and IL-1 beta, these cytokines were measured in whole blood samples stimulated with LPS (1 ng/mL) in the presence or absence of concentrations of the peptide. MEASUREMENTS AND MAIN RESULTS: Plasma alpha-MSH was low in early samples and gradually increased in patients who recovered but not in those who died. There was a negative correlation between plasma concentrations of alpha-MSH and TNF-alpha. In blood samples taken at early phases of sepsis syndrome, production of TNF-alpha was reduced relative to control values; such production increased in patients who recovered but not in those who died. Addition of alpha-MSH to LPS-stimulated whole blood samples inhibited production of TNF-alpha and IL-1beta in a concentration-dependent manner. CONCLUSIONS: In patients with systemic inflammation, there are substantial changes over time in plasma concentrations of alpha-MSH that are reduced in early phases of the disease. Reduction of this endogenous modulator of inflammation could be detrimental to the host. Addition of alpha-MSH to LPS-stimulated blood samples reduces production of cytokines involved in development of septic syndrome. This inhibition by alpha-MSH, a peptide that is beneficial in treatment of experimental models of sepsis, might therefore be useful to treat sepsis syndrome in humans. PMID- 10834687 TI - Metoclopramide for preventing pneumonia in critically ill patients receiving enteral tube feeding: a randomized controlled trial. AB - OBJECTIVE: To determine whether metoclopramide prevents nosocomial pneumonia in intensive care unit (ICU) patients receiving enteral feeding by a nasogastric tube. DESIGN: Prospective, randomized, controlled trial. SETTING: ICU of a university hospital. PATIENTS: A total of 305 consecutive patients requiring placement of a nasogastric tube for >24 hrs. INTERVENTIONS: Patients were randomized to receive either 10 mg of metoclopramide or placebo at 8-hr intervals through the nasogastric tube. MEASUREMENTS AND MAIN RESULTS: A total of 174 patients received placebo and 131 received metoclopramide. Baseline characteristics in the two treatment groups were comparable. Of the 305 patients, 46 developed nosocomial pneumonia, which was 24 patients (13.7%) in the placebo group and 22 (16.8%) in the metoclopramide group (p > .05). Patients in the placebo group developed pneumonia earlier than patients receiving metoclopramide (4.46+/-1.72 days [mean +/- SD[rsqb] after ICU admission compared with 5.95+/ 1.78 days; p = .006). Subgroup analysis showed that metoclopramide did not reduce the frequency rate of pneumonia in patients with tracheal intubation (19 [25.3%] of 75 patients receiving metoclopramide vs. 21 [21.2%] of 99 patients receiving placebo) or those receiving mechanical ventilation (17 [25.6%] of 58 patients receiving metoclopramide vs. 20 [29.3%] of 78 patients receiving placebo). The mortality rate also did not differ in the two treatments groups (56% in the metoclopramide group vs. 53% in the placebo group; p > .05). CONCLUSIONS: Although metoclopramide delayed the development of nosocomial pneumonia, it did not decrease its frequency rate and had no effect on the mortality rate in critically ill patients receiving nasogastric enteral feeding. PMID- 10834689 TI - Is protease inhibitor a choice for the treatment of pre- or mild disseminated intravascular coagulation? AB - OBJECTIVE: To investigate the effect of a protease inhibitor, gabexate mesylate, on patients with pre- or mild disseminated intravascular coagulation (DIC) in comparison with a control group receiving no anticoagulation therapy. DESIGN: Prospective, randomized, controlled study. SETTING: General intensive care unit at a general hospital. PATIENTS: Adult patients (40) with a DIC score between 6 and 8 (pre- or mild DIC). INTERVENTIONS: In 20 patients, gabexate mesylate (2 mg/kg/hr) was administered as 2 mL/hr in saline (treated group) and in another 20 patients, saline (2 mL/hr; control group) was administered during the study (7 days). MEASUREMENTS AND MAIN RESULTS: The following variables were determined at the time of admission to the intensive care unit before treatment and 1, 3, 5, and 7 days thereafter: platelet count, antithrombin III activity, serum or plasma concentrations of fibrinogen, fibrin degradation product, D-dimer, fibrin monomer, thrombin-antithrombin III complex, and plasmin-plasmin inhibitor complex, prothrombin time ratio, and DIC score. Two patients in the treated group and four in the control group were excluded from the study because they died during the study; therefore, 34 patients were analyzed. The measured variables of coagulation and fibrinolysis were not significantly different between the two groups, except for the D-dimer on day 3 (the treated group showed a higher concentration). D-dimer concentration and DIC score went down more quickly in the control group than the treated group, but not significantly. The mortality rate at 1 month was 40% (8 of 20) in the treated group and 35% (7 of 20) in the control group, without any differences between the two groups. CONCLUSIONS: In a limited number of patients (n = 34), gabexate mesylate (2 mg/kg/hr) could not inhibit coagulation or fibrinolysis and gabexate mesylate could not improve the DIC score or mortality rate in pre- or mild DIC. PMID- 10834688 TI - Prolonged use of heat and moisture exchangers does not affect device efficiency or frequency rate of nosocomial pneumonia. AB - OBJECTIVE: To determine whether use of a single heat and moisture exchanger (HME) for < or =120 hrs affects efficiency, resistance, level of bacterial colonization, frequency rate of nosocomial pneumonia, and cost compared with changing the HME every 24 hrs. DESIGN: Prospective, controlled, randomized, unblinded study. SETTING: Surgical intensive care unit at a university teaching hospital. PATIENTS: A total of 220 consecutive patients requiring mechanical ventilation for >48 hrs. INTERVENTIONS: Patients were randomized to one of three groups: a) hygroscopic HME (Aqua+) changed every 24 hrs (HHME-24); b) hydrophobic HME (Duration HME) changed every 120 hrs (HME-120); and c) hygroscopic HME (Aqua+) changed every 120 hrs (HHME-120). Devices in all groups could be changed at the discretion of the staff when signs of occlusion or increased resistance were identified. MEASUREMENTS AND MAIN RESULTS: Daily measurements of inspired gas temperature, inspired relative humidity, and device resistance were made. Additionally, daily cultures of the patient side of the device were accomplished. The frequency rate of nosocomial pneumonia was made by using clinical criteria. Ventilatory support variables, airway care, device costs, and clinical indicators of humidification efficiency (sputum volume, sputum efficiency) were also recorded. Prolonged use of both hygroscopic and hydrophobic devices did not diminish efficiency or increase resistance. There was no difference in the number of colony-forming units from device cultures over the 5-day period and no difference between colony-forming units in devices changed every 24 hrs compared with devices changed after 120 hrs. The average duration of use was 23+/-4 hrs in the HHME-24 group, 73+/-13 hrs in the HME-120 group, and 74+/-9 hrs in the HHME 120 group. Mean absolute humidity was greater for the hygroscopic devices (30.4+/ 1.1 mg of H2O/L) compared with the hydrophobic devices (27.8+/-1.3 mg of H2O/L). The frequency rate of nosocomial pneumonia was 8% (8:100) in the HHME-24 group, 8.3% (5:60) in the HME-120 group, and 6.6% (4:60) in the HHME-120 group. Pneumonia rates per 1000 ventilatory support days were 20:1000 in the HHME-24 group, 20.8:1000 in the HME-120 group, and 16.6:1000 in the HHME-120 group. Costs per day were $3.24 for the HHME-24 group, $2.98 for the HME-120 group, and $1.65 for the HHME-120 group. CONCLUSIONS: Changing the hydrophobic or hygroscopic HME after 3 days does not diminish efficiency, increase resistance, or alter bacterial colonization. The frequency rate of nosocomial pneumonia was also unchanged. Use of HMEs for >24 hrs, up to 72 hrs, is safe and cost effective. PMID- 10834690 TI - Adverse effects of high-dose epinephrine on cerebral blood flow during experimental cardiopulmonary resuscitation. AB - OBJECTIVE: To study the effects of high-dose epinephrine, compared with standard dose epinephrine, on the dynamics of superficial cortical cerebral blood flow as well as global cerebral oxygenation during experimental cardiopulmonary resuscitation. We hypothesized that high-dose epinephrine might be unable to improve cerebral blood flow during cardiopulmonary resuscitation as compared with standard-dose epinephrine. DESIGN: Randomized controlled study. SETTING: University hospital research laboratory. SUBJECTS: A total of 20 male anesthetized piglets. INTERVENTIONS: Ventricular fibrillation was induced. A nonintervention interval of 8 mins was followed by open-chest cardiopulmonary resuscitation. The animals were randomized to receive repeated bolus injections of either 20 microg/kg (standard-dose group, n = 10) or 200 microg/kg (high-dose group, n = 10) of epinephrine. MEASUREMENTS AND MAIN RESULTS: Focal cortical cerebral blood flow was measured continuously by using laser Doppler flowmetry. The duration of blood flow increase was significantly shorter in the high-dose group after the second dose of epinephrine. In the high-dose group there was also a consistent tendency for lower peak levels and shorter duration of flow increase in response to repeated bolus doses of epinephrine. Cerebral oxygen extraction ratio was significantly lower in the high-dose group after administration of epinephrine. CONCLUSIONS: Repeated bolus doses of epinephrine 200 microg/kg, as compared with 20 microg/kg, do not improve superficial cortical cerebral blood flow during experimental open-chest cardiopulmonary resuscitation. High-dose epinephrine appears to induce vasoconstriction of cortical cerebral blood vessels resulting in redistribution of blood flow from superficial cortex. This might be one explanation for the failure of high-dose epinephrine to improve overall outcome in clinical trials. PMID- 10834691 TI - Desmethyl tirilazad improves neurologic function after hypoxic ischemic brain injury in piglets. AB - OBJECTIVE: Desmethyl tirilazad is a lipid-soluble free radical quencher. Deferoxamine reduces free radicals by chelating iron and reducing hydroxyl formation. Free radical inhibitors have shown promise in several hypoxic ischemic brain injury models, and we wished to see if this work could be extended to our newborn piglet model. DESIGN: Randomized controlled trial. SUBJECTS: Piglets (0 to 3 days old). INTERVENTION: Carotid snares and arterial and venous catheters were placed under 1.5% isoflurane anesthesia. In Experiment 1, piglets were randomly assigned to receive either 3 mg/kg desmethyl tirilazad or vehicle at -15 and 90 mins. In Experiment 2, piglets were randomly assigned to receive either 20 mg/kg desmethyl tirilazad at -15 mins followed by 8 mg/kg/hr for 90 mins or 100 mg/kg deferoxamine at -15 mins or vehicle. At time 0, both carotid arteries were clamped and blood was withdrawn to reduce the blood pressure to two-thirds normal. At 15 mins, inspired oxygen was reduced to 6%. At 30 mins, the carotid snares were released, the withdrawn blood was reinfused, and the oxygen was switched to 100%. On the third day after the hypoxic ischemic injury, the animals were killed by perfusing their brains with 10% formalin. We tested the timing of lipid peroxidation and inhibition of lipid peroxidation by these agents by freezing the brains of a subset of pigs in liquid nitrogen. MEASUREMENTS: Neurologic examination and brain pathology were scored by blinded observers. Thiobarbituric acid-reactive substance and oxidized and reduced glutathione were measured on frozen brains. MAIN RESULTS: Desmethyl tirilazad (20 mg/kg) and 100 mg/kg deferoxamine inhibit lipid peroxidation. Desmethyl tirilazad (20 mg/kg) improves neurologic exam, but 3 mg/kg Desmethyl tirilazad or 100 mg/kg deferoxamine does not. Neither desmethyl tirilazad nor deferoxamine improves pathologic results. CONCLUSIONS: High-dose desmethyl tirilazad improves neurologic function after hypoxic ischemic brain injury in the newborn piglet. PMID- 10834692 TI - Endotoxin induces a dose-dependent myocardial cross-tolerance to ischemia reperfusion injury. AB - OBJECTIVE: To test whether or not endotoxin induces a dose-dependent reduction of myocardial contractile dysfunction after a standardized period of myocardial ischemia and reperfusion and whether nitric oxide is involved in this form of myocardial protection. DESIGN: Prospective, randomized, controlled animal study. SETTING: University research laboratory. SUBJECTS: Twenty-five male Sprague Dawley rats. INTERVENTIONS: After anesthesia, the left carotid artery was cannulated under sterile conditions and animals were allowed to recover from surgery for 12 hrs. Sterile saline or increasing doses (2.5, 5, or 10 mg/kg body weight) of endotoxin (Escherichia coli O26:B6; Sigma, Mississauga, Ontario, Canada) were given intravenously (1 mL over 5 mins). In some rats, diaspirin crosslinked hemoglobin (200 mg/kg) was infused 6 hrs and 60 min before endotoxin infusion (10 mg/kg). Hearts were rapidly excised for retrograde perfusion through the ascending aorta (Langendorff apparatus) 6 hrs later. After baseline data collection, hearts were subjected to global ischemia (30 mins, 37 degrees C [98.6 degrees F]), followed by 30 mins of reperfusion. MEASUREMENTS AND MAIN RESULTS: Physiologic variables were recorded 6 hrs after saline and endotoxin infusion. Baseline myocardial systolic contractility and diastolic compliance were assessed, respectively, by left ventricular developed pressure (LVDP) and left ventricular (LV) volume-preload relationships. After 30 min of reperfusion, LVDP recovery and left ventricular end-diastolic pressure were measured. Endotoxin induced LV systolic contractile depression, irrespective of the dose of endotoxin administered. LV diastolic dysfunction varied between different doses of endotoxin administered. On reperfusion, endotoxin produced a dose-dependent improvement of postischemic LVDP recovery: 30+/-6% in sham, 78+/-9% in 2.5 mg/kg, 93+/-8% in 5 mg/kg, and 107+/-10% in 10 mg/kg endotoxin heart. In rats treated with 10 mg/kg endotoxin, diaspirin-crosslinked hemoglobin pretreatment abrogated endotoxin-induced postischemic LVDP recovery improvement (105+/-10% vs. 43+/-7%, p = .01). CONCLUSION: Sublethal doses of endotoxin induce in a dose-dependent manner a delayed form of myocardial protection against ischemia. Although free cell hemoglobin solution abrogates this endotoxin-induced cross-tolerance, we propose that possible mechanisms involved in this form of myocardial protection include nitric oxide pathway activation. PMID- 10834693 TI - Epinephrine application via an endotracheal airway and via the Combitube in esophageal position. AB - OBJECTIVE: To compare plasma concentrations and cardiovascular effects of epinephrine after application via a conventional endotracheal airway and via the esophageal lumen of a new emergency airway, the esophageal tracheal Combitube. DESIGN: Prospective, randomized study. SETTING: Center for Biomedical Research, University of Vienna. SUBJECTS: Fourteen juvenile swine received either an endotracheal tube (Group A) or a Combitube in esophageal position (Group B). INTERVENTIONS: In Part I of the study, epinephrine was administered during spontaneous beating of the heart; in Part II, epinephrine was administered during cardiopulmonary resuscitation, using a ten-fold higher dosage in Group B, respectively. MEASUREMENTS: Plasma epinephrine levels were measured 1, 2, 3, 5, 7, 10, 15, and 30 mins after application. Systolic arterial blood pressure and cardiac output in Part I, and end-tidal CO2 and coronary perfusion pressure in Part II were recorded. MAIN RESULTS: In Part I, increased levels of plasma epinephrine and systolic arterial pressure were maintained significantly longer in Group B when compared with Group A. In Part II, no significant differences between the groups were found with regard to plasma epinephrine levels and hemodynamic variables. CONCLUSION: Epinephrine applied via the esophageal lumen of the Combitube in a ten-fold higher dosage has similar effects on plasma epinephrine levels and hemodynamic variables compared to endotracheal administration. PMID- 10834694 TI - At surfactant deficiency, application of "the open lung concept" prevents protein leakage and attenuates changes in lung mechanics. AB - OBJECTIVE: To evaluate whether mechanical ventilation using "the open lung concept" during surfactant depletion can attenuate the deterioration in pulmonary function. DESIGN: Experimental, comparative study. SETTING: Research laboratory of a large university. SUBJECTS: Eighteen adult male Sprague-Dawley rats, weighing 280-340 g. INTERVENTIONS: Twelve rats were anesthetized, mechanically ventilated with 100% oxygen, and randomly divided into two groups (n = 6 each). The open lung group underwent six saline lavages at different ventilator settings that prevented alveolar collapse. The settings (expressed as frequency/peak inspiratory pressure/positive end-expiratory pressure/inspiratory:expiratory ratio) were 30/26/6/1:2 during the first lavage, 100/27/10/1:1 during the next two lavages, and 100/33/15/1:1 during the last three lavages and during the remaining ventilation period. The ventilated control group underwent six saline lavages with settings at 30/26/6/1:2. After the lavages, peak inspiratory pressure and positive end-expiratory pressure were increased in this group by 2 cm H2O each for the remaining study period. An additional group of six animals were killed immediately after induction of anesthesia and served as healthy controls. Blood gases were measured before lavage, immediately after the last lavage, and thereafter hourly. At the end of the 4-hr study period, we constructed pressure-volume curves from which we determined total lung capacity at a distending pressure of 35 cm H2O (TLC35). Subsequently, total lung volume at a distending pressure of 5 cm H2O (V5) was determined, followed by bronchoalveolar lavage. RESULTS: In the ventilated control group, PaO2, V5, and TLC35 were significantly decreased and protein concentration of bronchoalveolar lavage was significantly increased compared with the healthy control group. In the open lung group, PaO2 did not decrease after the lavage procedure, and V5, TLC35, and the protein concentration of bronchoalveolar lavage were comparable with the healthy controls. CONCLUSION: We conclude that application of the open lung concept during surfactant depletion attenuates deterioration in pulmonary function. PMID- 10834695 TI - A comparison of intratracheal pulmonary ventilation to conventional ventilation in a surfactant deficient animal model. AB - OBJECTIVE: To compare intratracheal pulmonary ventilation (ITPV) with conventional ventilation in a rabbit model of surfactant deficiency. DESIGN: A prospective randomized animal study. SETTING: The Children's National Medical Center Research Animal Facility in Washington, DC. SUBJECTS: Adult male New Zealand white rabbits (n = 20), weighing 1.4-4.2 kg. INTERVENTIONS: After anesthesia and catheter placement, rabbits were tracheotomized, paralyzed, and placed on the conventional ventilator. We determined pulmonary functions at baseline. We washed surfactant out of the lungs by using serial bronchoalveolar lavages. Pulmonary function studies were determined after completion of the bronchoalveolar lavages and were used as an indication of severity of lung injury. Animals were randomized into two groups: We placed ten animals on ITPV, using the ITPV reverse thruster catheter designed by Kolobow and a prototype ITPV ventilator designed at Children's National Medical Center; we placed ten animals on conventional ventilation using the Sechrist iv-100 ventilator. Arterial blood gases were drawn every 15 mins, and the ventilator settings were adjusted to the minimal level that would maintain arterial blood gases in the following ranges: pH 7.35-7.45, PaCO2 30-40 torr (3.995.33 kPa), PaO2 50-70 torr (6.66-9.33 kPa). Animals were ventilated with the randomized ventilation techniques for 4 hrs. MEASUREMENTS AND MAIN RESULTS: Peak inspiratory pressure, mean airway pressure, and positive end-expiratory pressure were measured at the distal end of the endotracheal tube. We recorded these variables plus respiratory rate at baseline and every 30 mins for a total of 4 hrs of ventilation. Lung compliance did not differ between groups at the postlavage study period (ITPV, 0.56+/-0.13 mL/cm H2O/kg; conventional 0.49+/-0.15 mL/cm H2O/kg). At the end of the 4 hr study period, peak inspiratory pressure (ITPV, 26.2+/-4.6 cm H2O; conventional, 32.4+/ 5.04 cm H2O, p = .007) and positive end-expiratory pressure (ITPV, 3.9+/-1.96 cm H2O; conventional, 6.3+/-1.42 cm H2O, p = .005) were lower in the ITPV ventilation group. Peak inspiratory pressure was significantly lower in the ITPV group by 2 hrs into the study. CONCLUSION: In this model of surfactant deficiency lung injury, ventilation and oxygenation were achieved at significantly lower ventilator settings using ITPV compared with conventional ventilation. Long-term studies are needed to determine whether this reduction in ventilation is maintained, and if so, if lung injury is reduced. PMID- 10834696 TI - Effects of partial liquid ventilation with perfluorodecalin in the juvenile rabbit lung after saline injury. AB - OBJECTIVE: To evaluate the feasibility of using the perfluorochemical, perfluorodecalin, for partial liquid ventilation (PLV) with respect to gas exchange and lung mechanics in normal and saline-injured lungs of juvenile rabbits. DESIGN: Experimental, prospective, randomized, controlled study. SETTING: Physiology laboratory at a university medical school. SUBJECTS: Seventeen juvenile rabbits assigned to three groups. INTERVENTIONS: The conventional mechanical ventilation (CMV)-injury group (n = 5) was treated with CMV after establishing a lung injury; the PLV-injury group (n = 6) was treated with PLV after lung injury; and the PLV-healthy group (n = 6) was supported with PLV without lung injury. Lung injury was created by repeated saline lung lavages. PLV-treated animals received a single dose of intratracheal perfluorodecalin at a volume equal to the measured preinjury gas functional residual capacity (functional residual capacity = 18.6+/-1.5 [SEM] mL/kg). MEASUREMENTS AND MAIN RESULTS: Sequential measurements of total respiratory compliance and arterial blood chemistries were performed in all groups. Oxygenation index (OI) and ventilation efficiency index were calculated. After lung injury, there was a significant (p < .05) decrease in PaO2, total respiratory compliance, and ventilation efficiency index and an increase in OI and PaCO2. In the PLV-injury group, PLV significantly (p < .05) improved PaO2 (+60%) and OI (-33%) over time. Compliance was significantly (p < .05) higher (90%) than in the CMV-injury group over time. CONCLUSIONS: These results demonstrate that PLV with perfluorodecalin improved oxygenation and increased respiratory compliance in the saline-injured rabbit lung. In addition, similar to the effects of several other perfluorochemical liquids on normal lungs, pulmonary administration of perfluorodecalin was associated with a small impairment in gas exchange and a significant decrease in lung compliance in the juvenile rabbit model. PMID- 10834697 TI - The effect of heat on cytokine production in rat endotoxemia. AB - OBJECTIVE: To determine whether heat stress protects the endotoxemic rat by up regulation of the counterinflammatory cytokine interleukin (IL)-10, thereby attenuating the inflammatory response. DESIGN: A total of 16 rats were assigned to either the heat stress group (n = 8) or the control group (n = 8). The heat stress group was warmed to a temperature of >42 degrees C (107.6 degrees F) rectally for 10-15 mins; 20 hrs later, all rats were intubated, paralyzed, and ventilated. After jugular venous and arterial catheterization, endotoxin was given intravenously. Arterial blood was removed at 0, 2, 4, and 5 hrs for blood gases, tumor necrosis factor (TNF)-alpha, nitric oxide metabolites (NO), IL-10, and macrophage inflammatory protein (MIP)-2. The alveolar macrophages were removed, counted, and then incubated for 24 hrs. The supernatant was analyzed for TNF-alpha, NO, IL-10, and MIP-2. SETTING: University research laboratory. SUBJECTS: Male Sprague-Dawley rats (n = 16). INTERVENTIONS: Administration of heat before endotoxin infusion. MEASUREMENTS AND MAIN RESULTS: Alveolar-arterial oxygen gradient was lower in the heat stress group at 4 and 5 hrs after endotoxemia. Plasma and alveolar macrophage supernatant concentrations of TNF alpha, NO, and IL-10 were not affected by heat. Plasma and alveolar macrophage supernatant MIP-2 concentrations were higher in endotoxemic rats receiving heat pretreatment compared with controls. CONCLUSIONS: Our study demonstrates that heat leads to pulmonary protection of short duration in severe endotoxemia. This protection was not mediated by plasma TNF-alpha, IL-10, or NO. Contrary to our hypothesis, pretreatment with heat increased rather than decreased the plasma MIP 2 concentration and alveolar macrophage production of MIP-2 in endotoxemia. The mechanism of heat-conferred pulmonary protection in endotoxemia remains unclear. Alveolar macrophages do not produce IL-10 in endotoxemia. The increased MIP-2 production by heated alveolar macrophages was not attributable to alterations in production of either TNF-alpha or IL-10. The significance of increased MIP-2 by endotoxin-exposed alveolar macrophages in heated rats is unknown. PMID- 10834698 TI - New clinically relevant sheep model of severe respiratory failure secondary to combined smoke inhalation/cutaneous flame burn injury. AB - OBJECTIVES: To develop a predictable, dose-dependent, clinically relevant model of severe respiratory failure associated with a 40% total body surface area, full thickness (third-degree) cutaneous flame burn and smoke inhalation injury in adult sheep. DESIGN: Model development. SETTING: Research laboratory. SUBJECTS: Adult female sheep (n = 22). INTERVENTIONS: Animals were divided into three groups, determined by the number of smoke breaths administered (24, 36, 48) for a graded inhalation injury. The smoke was insufflated into a tracheostomy with a modified bee smoker at airway temperatures <40 degrees C. All animals concurrently received a 40% total body surface area (third-degree) cutaneous flame burn to the body (flanks). After injury, the animals were placed on volume controlled ventilation to achieve PaO2 >60 mm Hg and PaCO2 <40 mm Hg. Arterial blood gases and ventilator settings were monitored every 6 hrs postinjury for up to 7 days. MEASUREMENTS AND MAIN RESULTS: All animals survived the induction of injury. In the 24 smoke breath/40% total body surface area burn (24/40) group, PaO2/F(IO2) never decreased below 300, and peak inspiratory pressure was consistently <14 cm H2O with normal arterial blood gases throughout the observation period. With 36 smoke breaths/40% total body surface area burn (36/40) (n = 7), all animals had PaO2/F(IO2) of <200 and peak inspiratory pressure of 26 cm H2O within 40-48 hrs, as 30% died during the study period. With 48 smoke breaths/40% total body surface area burn (48/40) (n = 12), all animals developed respiratory distress syndrome (RDS) in 24-30 hrs, but none survived the experimental period. CONCLUSIONS: Development of RDS by smoke and cutaneous flame bum injury depends on smoke inhalation dose. A combination of 36 breaths of smoke and a 40% total body surface area (third-degree) cutaneous flame burn injury can induce severe RDS (PaO2/F(IO2) <200) within 40-48 hrs to allow evaluation of various treatment modalities of RDS. PMID- 10834699 TI - Trauma and inflammation modulate lymphocyte localization in vivo: quantitation of tissue entry and retention using indium-111-labeled lymphocytes. AB - OBJECTIVE: Determine the in vivo localization pattern of indium-111-labeled lymphocytes after a standardized extremity injury or standardized laparotomy and after sterile inflammation of the central nervous system. DESIGN: Prospective animal study with concurrent controls. SETTING: Animal research laboratory. SUBJECTS: Male Lewis rats weighing 150-175 g. INTERVENTIONS: Indium-111-labeled splenic lymphocytes were injected into animals after a standardized hind limb trauma or laparotomy and after induction of sterile central nervous system inflammation. MEASUREMENTS AND MAIN RESULTS: Lymphoid and non-lymphoid organs were removed at fixed intervals after lymphocyte injection and the proportion of injected lymphocytes/gram of tissue was determined using a quantitative radionuclide calculation. Results from treated animals were compared with results from untreated control animals. Muscle injury caused early localization of lymphocytes to injured hind limbs, liver, and spleen compared with controls, whereas laparotomy decreased lymphocyte localization in the thymus and colon. Encephalitis increased localization to the central nervous system with no effect on other tissues. CONCLUSIONS: These results identify a sensitive method to track in vivo leukocyte localization and specifically demonstrate that lymphocyte localization is altered in both traumatic and nontraumatic models of inflammation. PMID- 10834700 TI - Development and application of a double-piston configured, total-liquid ventilatory support device. AB - OBJECTIVE: Perfluorocarbon liquid ventilation has been shown to enhance pulmonary mechanics and gas exchange in the setting of respiratory failure. To optimize the total liquid ventilation process, we developed a volume-limited, time-cycled liquid ventilatory support, consisting of an electrically actuated, microprocessor-controlled, double-cylinder, piston pump with two separate limbs for active inspiration and expiration. DESIGN: Prospective, controlled, animal laboratory study, involving sequential application of conventional gas ventilation, partial ventilation (PLV), and total liquid ventilation (TLV). SETTING: Research facility at a university medical center. SUBJECTS: A total of 12 normal adult New Zealand rabbits weighing 3.25+/-0.1 kg. INTERVENTIONS: Anesthestized rabbits were supported with gas ventilation for 30 mins (respiratory rate, 20 cycles/min; peak inspiratory pressure, 15 cm H2O; end expiratory pressure, 5 cm H2O), then PLV was established with perflubron (12 mL/kg). After 15 mins, TLV was instituted (tidal volume, 18 mL/kg; respiratory rate, 7 cycles/min; inspiratory/expiratory ratio, 1:2 cycles/min). After 4 hrs of TLV, PLV was re-established. MEASUREMENTS AND MAIN RESULTS: Of 12 animals, nine survived the 4-hr TLV period. During TLV, mean values +/- SEM were as follows: PaO2, 363+/-30 torr; PaCO2, 39+/-1.5 torr; pH, 7.39+/-0.01; static peak inspiratory pressure, 13.2+/-0.2 cm H2O; static endexpiratory pressure, 5.5+/-0.1 cm H2O. No significant changes were observed. When compared with gas ventilation and PLV, significant increases occurred in mean arterial pressure (62.4+/-3.5 torr vs. 74.0+/-1.2 torr) and central venous pressure (5.6+/-0.7 cm H2O vs. 7.8+/ 0.2 cm H2O) (p < .05). CONCLUSIONS: Total liquid ventilation can be performed successfully utilizing piston pumps with active expiration. Considering the enhanced flow profiles, this device configuration provides advantages over others. PMID- 10834701 TI - Fish oil-supplemented parenteral diets normalize splanchnic blood flow and improve killing of translocated bacteria in a low-dose endotoxin rat model. AB - OBJECTIVE: Intestinal ischemia decreases barrier function of the gut and enhances translocation of bacteria and toxins. Several studies indicate that fish oil can modulate prostaglandin formation and thus, regional blood flow and immune function. This study was performed to determine the effects of parenteral diets with omega-3 fatty acids on microcirculation and barrier function of the gut.0 DESIGN: Prospective, randomized, controlled animal study. SETTING: University laboratory. SUBJECTS: A total of 64 male Sprague-Dawley CD rats. INTERVENTIONS AND MEASUREMENTS: For 48 hrs, eight groups of eight rats each received total parenteral nutrition with four different types of lipids. The source of fat in group L was soybean oil only and in group L-M a mixture of soybean oil and medium chain triglycerides. In groups FO-20 and FO-40, 20% or 40%, respectively, of the soybean oil in group L-M was replaced by fish oil. The other four groups received an additional continuous infusion of endotoxin (0.1 mg/100 g body weight per day) for the last 24 hrs. Blood flow was measured with microspheres, and translocation was determined by microbiological methods and instillation of radioactive-marked bacteria into the gut. MAIN RESULTS: In the animals without fish oil, the endotoxin application reduced the blood flow to the intestine approximately 25%. Animals with fish oil in their diets showed normal values. Translocation of gut bacteria was increased significantly in all endotoxin groups. However, less viable bacteria could be detected in the animals with fish oil diets in their mesenteric lymph nodes and livers. CONCLUSIONS: In this model, diets enriched with fish oil abolish the endotoxin-induced decrease of nutritive blood flow to the gut and ameliorate the bactericidal defense of the splanchnic region. The lower count of viable bacteria in the fish oil groups is more related to an improved killing of translocated bacteria than a reduction of the translocation rate. PMID- 10834702 TI - Acute lung injury in two experimental models of acute pancreatitis: infusion of saline or sodium taurocholate into the pancreatic duct. AB - OBJECTIVE: To compare acute pulmonary changes secondary to sodium taurocholate hemorrhagic pancreatitis with those changes secondary to a less severe pancreatitis induced by saline infusion into the biliopancreatic duct. DESIGN: Prospective, randomized controlled trial. SETTING: University pulmonary laboratory. SUBJECTS: A total of 110 male Wistar rats. INTERVENTIONS: Pancreatitis was induced by either 0.5 mL of a 4% solution of sodium taurocholate (TAU group) or 0.5 mL of normal saline (SAL group) injection into the biliopancreatic duct. Data were compared with data from control (sham-operated) animals (SHAM group). MEASUREMENTS AND MAIN RESULTS: The severity of pancreatic and pulmonary injuries was evaluated 1, 3, and 8 days after the induction of acute pancreatitis by morphometric and pulmonary mechanical studies. Biliopancreatic duct pressure was measured during infusion of solutions in SAL and TAU groups. SAL and TAU groups developed an intense necrohemorrhagic pancreatitis on day 1 without differences in biliopancreatic duct pressures (134.0+/-45.1 cm H2O vs. 123.3+/-23.4 cm H2O). Acute pancreatic lesions were still intense on day 3 in the TAU group only. Pulmonary resistance in SAL and TAU groups was significantly greater than in the SHAM group on day 3 only. On day 1, there was an increase in intraalveolar edema in both groups (p < .02). There was an increase in polymorphonuclear cells in alveolar septa on day 1 only in the TAU group (p < .001). In contrast, both experimental groups presented greater values of PMN cells on day 8 compared with the SHAM group (p < .001). Both groups with pancreatitis showed an increase in alveolar distention and collapse on day 1 that persisted only in the TAU group on days 3 and 8. No deaths were observed in the control (SHAM) group. In contrast, the SAL group had lower mortality than the TAU group in the first two days (17% and 52%, respectively, p = .03). CONCLUSION: High-pressure infusion of normal saline into the biliopancreatic duct of rats results in significant pancreatic and lung alterations. These changes are worse in the presence of sodium taurocholate. PMID- 10834703 TI - Left ventricular myocardial adenosine triphosphate changes during reperfusion of ventricular fibrillation: the influence of flow and epinephrine. AB - OBJECTIVE: To determine whether epinephrine in combination with high flow worsens left ventricular (LV) myocardial high-energy phosphate stores during reperfusion of ischemic ventricular fibrillation (VF). DESIGN: Blinded, prospective block randomized, placebo controlled study. SETTING: University medical center research laboratory. SUBJECTS: A total of 22 mixed breed swine weighing 22.0+/-3.3 kg (SD). INTERVENTIONS: Open-chest swine, anesthetized with alpha-chloralose, underwent 10 mins of nonperfused VF followed by reperfusion with cardiopulmonary bypass for 90 mins and then defibrillation. Animals were block randomized to four groups for reperfusion: Group 1 (n = 5), high flow (100 mL/kg/min) and epinephrine (2.5 microg/kg/min); Group 2 (n = 5), high flow and placebo; Group 3 (n = 6), low flow (30 mL/kg/min) and epinephrine; and Group 4 (n = 6), low flow and placebo. MEASUREMENTS AND MAIN RESULTS: In vivo LV creatine phosphate (CP) and adenosine triphosphate (ATP) were determined using whole wall and spatially localized 31P NMR spectroscopy at 4.7 Tesla. During perfusion of the fibrillating myocardium, epinephrine significantly increased aortic pressure (p < .05) and improved defibrillation rates (p < .01). ATP levels during reperfusion were significantly decreased within all groups compared with baseline. There were no differences in ATP levels between groups. High flow, independent of epinephrine, was associated with increased preservation of ATP (p < .05), increased CP/ATP ratios (p < .02) in all layers of the LV wall, and decreased aortic and cardiac vein lactates (p < .001). CONCLUSIONS: Epinephrine, in combination with flow higher than standard cardiopulmonary resuscitation flows, increased perfusion pressure and defibrillation rates, but did not significantly alter myocardial ATP during VF reperfusion in the in vivo heart Reperfusion flow, independent of epinephrine, is a critical determinant of myocardial ATP preservation. PMID- 10834704 TI - Hydrogen peroxide induces midzonal heat shock protein 72 and apoptosis in sinusoidal endothelial cells of hypoxic rat liver. AB - OBJECTIVE: To investigate the heat shock protein (HSP) 72 expression and apoptosis induced by hydrogen peroxide in hypoxic rat liver. DESIGN: Prospective control study using the isolated rat liver. SETTING: Animal research facility. SUBJECTS: Fasted, pathogen-free specific, male Sprague-Dawley rats. INTERVENTIONS: A low-flow hypoxia model was made by reducing an afferent pressure from 10 to 2.5 cm H2O, and by perfusing the isolated rat liver for 2 hrs. MEASUREMENT AND MAIN RESULTS: We investigated the hydrogen peroxide production by using the 2'-7' dichlorofluorescein image, the induction of HSP 72 by using immunohistochemistry, and apoptosis by using terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick end-labeling method in the low flow hypoxic rat liver. In low-flow hypoxia, hydrogen peroxide production, HSP 72 expression, and apoptosis were induced in the midzone of rat liver. Prevalence of HSP 72 expression was higher in the sinusoidal endothelial cells (SEC) than in the hepatocytes. All apoptotic cells were SEC with expression of HSP 72. Hydrogen peroxide was derived from hepatocytes. Pretreatment with the specific xanthine oxidase inhibitor, sodium(-)-8-(3-methoxy-phenylsulfinylphenyl) pyrazolo [1,5-a] 1,3,5-triazine-4-olate monohydrate significantly attenuated hydrogen peroxide production, HSP 72 expression, and apoptosis of SEC in the midzone. CONCLUSION: Xanthine oxidase-dependent hydrogen peroxide induces midzonal and SEC-dominant HSP 72 expression and apoptosis in hypoxic rat liver. PMID- 10834705 TI - Endothelin-1 impairs neutrophil respiratory burst and elimination of Escherichia coli in rabbits. AB - OBJECTIVE: During systemic inflammation, elevated levels of endothelin (ET)-1 have been reported. The aim of this study was to investigate the effects of ET-1 on neutrophil (PMN) respiratory burst, phagocytosis, and elimination of Escherichia coli from blood and tissues. DESIGN: Prospective, randomized, controlled trial. SETTING: Experimental laboratory in a university hospital. SUBJECTS: A total of 18 female chinchilla rabbits. INTERVENTIONS: To quantify the clearance process, defined numbers (10(8) colony-forming units) of E. coli were injected intravenously into anesthetized rabbits, 60 mins after onset of continuous 0.2 microg/kg/min ET-1 administration (n = 9) and after saline infusion (control group, n = 9), respectively. To evaluate potential effects of ET-1 on bacterial elimination and killing, blood clearance of E. coli and colonization of different organs were investigated. MEASUREMENTS: Variables monitored were neutrophil respiratory burst and phagocytosis activity, rates of bacterial elimination from the blood, arterial blood pressure, blood gases, serum lactate concentrations, and nitrite and nitrate levels. The animals were killed 3 hrs after bacterial injection and tissue samples of liver, kidney, spleen, and lung were collected for bacterial counts. MAIN RESULTS: Compared with the control group, ET-1 significantly impaired PMN respiratory burst (p < .05) and prolonged elimination of injected E. coli from the blood (p < .01), whereas phagocytosis functions remained unaltered. The reduced PMN burst activity after ET-1 was associated with a higher bacterial colonization of all organs (lung, p < .01; spleen, p < .05). Endothelin-1 induced increases in mean arterial pressure (p < .01) and serum lactate concentrations, whereas nitrite and nitrate levels remained unaltered. CONCLUSION: Endothelin-1 impairs respiratory burst and bacterial clearance from the blood and tissue. Thus, elevated levels of ET-1 during sepsis could induce organ hypoperfusion and cause disturbances in immune functions, increasing the risk of bacterial infections. PMID- 10834706 TI - Effects of partial liquid ventilation on regional pulmonary blood flow distribution of isolated rabbit lungs. AB - OBJECTIVE: Partial liquid ventilation with perfluorocarbons may increase alveolar hydrostatic transmural pressure and may result in a redistribution of pulmonary blood flow from dependent to nondependent lung regions. To test this hypothesis under controlled study conditions, we determined intrapulmonary blood flow distributions during gas and perfluorocarbon ventilation in isolated rabbit lungs. DESIGN: Controlled animal study with an ex vivo isolated lung preparation. SETTING: Research laboratory for Experimental Anesthesiology at the Heinrich Heine-University of Dusseldorf. SUBJECTS: New Zealand White rabbits. INTERVENTIONS: The lungs were perfused with autologous blood at constant flow (150 mL/min) and ventilated with 5% C(O2) in air (positive end-expiratory pressure, 2 cm H2O; tidal volume, 10 mL/kg body weight; respiratory rate, 30 breaths/ min) without and with perfluorocarbon administered intratracheally (15 mL/kg). MEASUREMENTS AND MAIN RESULTS: Regional lung perfusion was measured with colored microspheres in apical, central, peripheral, and basal samples before and after bronchial instillation of perfluorocarbons. Compared with gas ventilation, intrapulmonary blood flow during perfluorocarbon ventilation was higher in apical samples (49.4+/-8.6 mL/min/g vs. 38.3+/-6.8 mL/min/g dry weight; p = .03) and lower in basal samples (22.2+/-5.1 mL/min/g vs. 39.9+/-8.2 mL/min/g; p = .04). CONCLUSIONS: Our findings suggest that during partial liquid ventilation, intrapulmonary blood flow is redistributed toward less-dependent lung regions. (Crit Care Med 2000; 28:1522-1525) PMID- 10834707 TI - Coupled plasma filtration-adsorption in a rabbit model of endotoxic shock. AB - OBJECTIVE: To test the hypothesis that nonselective adsorption by a hydrophobic resin of cytokines and other proinflammatory mediators could improve 72-hr survival in a rabbit model of endotoxic shock. DESIGN: Prospective, randomized, controlled animal trial. SETTING: Animal care facility at a research institution. SUBJECTS: A total of 109 New Zealand white male rabbits. INTERVENTIONS: Anesthetized rabbits were cannulated with indwelling femoral arterial and venous lines. Septic shock was induced by a single intravenous injection of Escherichia coli lipopolysaccharide. The dose was experimentally assessed in 40 rabbits receiving 1.0, 0.5, 0.1, and 0.05 mg/kg body weight to determine LD80 at 72 hrs. Extracorporeal circulation consisted of plasma filtration coupled with passage of the plasma filtrate through a hydrophobic sorbent and reinfusion into the venous line. The extracorporeal treatment lasted for 3 hrs. Rabbits injected with endotoxin (0.05 mg/kg) were submitted to plasma filtration with (19 rabbits) or without (20 rabbits) sorbent adsorption. As controls, rabbits injected with vehicle alone were treated with plasma filtration (ten rabbits) or without (ten rabbits) sorbent adsorption. Ten rabbits were monitored under anesthesia to determine basal survival. MEASUREMENTS AND MAIN RESULTS: Plasma concentrations of endotoxin, bioactive tumor necrosis factor, resin-adsorbed platelet-activating factor, mean arterial pressure, base excess, and white cell count were assessed and a global severity score was established. At 72 hrs, cumulative survival was significantly (p = .0041) improved in septic rabbits treated with coupled plasma filtration-adsorption. Circulating tumor necrosis factor bioactivity remained similar in control and treated rabbits. Biologically significant amounts of platelet activating factor were eluted from the sorbent during the entire treatment time. The severity score inversely correlated with survival (p < .001). CONCLUSIONS: Coupled plasma filtration-adsorption improved survival in a rabbit model of endotoxic shock. Coupled plasma filtration-adsorption may be an extracorporeal treatment capable of removing structurally different inflammatory mediators associated with sepsis. PMID- 10834708 TI - Is prostacyclin responsible for producing the hyperdynamic response during early sepsis? AB - OBJECTIVE: Although polymicrobial sepsis is characterized by an early hyperdynamic phase (2-10 hrs after cecal ligation and puncture [CLP]), followed by a late hypodynamic phase (20 hrs after CLP), it remains unknown whether prostacyclin or prostaglandin I2 (PGI2) plays a significant role in modulating the hyperdynamic state during early sepsis. The aim of this study was to determine whether inhibition of PGI2 synthesis prevents the occurrence of the hyperdynamic response during early sepsis. DESIGN: Prospective, controlled animal study. SETTING: A university research laboratory. SUBJECTS: Adult male Sprague Dawley rats were subjected to sepsis by CLP. INTERVENTIONS AND MEASUREMENTS: Blood samples were collected at 2, 5, 10, or 20 hrs after CLP, and plasma concentrations of PGI2, in the form of its stable product 6-keto-PGF1alpha, were measured by radioimmunoassay. In additional studies, a PGI2 synthase inhibitor, tranylcypromine, was administered subcutaneously at the time of CLP and again at 3 hrs after CLP. At 5 hrs after the onset of sepsis, the maximal rates of the left ventricular pressure rise (+dP/dtmax) and fall (-dP/dtmax) were determined by an in vivo heart performance analyzer. Microvascular blood flow in the liver, small intestine, and spleen was assessed by laser Doppler flowmetry. MAIN RESULTS: Plasma concentrations of 6-keto-PGF1alpha increased significantly at 2 20 hrs after CLP. At 5 hrs after the onset of sepsis, +/-dP/dt(max) and microvascular blood flow in the tested tissues increased significantly. Inhibition of PGI2 synthase activity did not prevent the occurrence of hypercardiovascular responses under such conditions. Moreover, the administration of tranylcypromine significantly reduced circulating concentrations of 6-keto PGF1alpha at 5 hrs after CLP. CONCLUSIONS: Because inhibition of PGI2 production did not prevent the occurrence of the hyperdynamic and hypercardiovascular response during the early stage of sepsis, mediators other than PGI2 appear to play a major role in producing the hyperdynamic response under such conditions. PMID- 10834709 TI - Lisofylline ameliorates intestinal and hepatic injury induced by hemorrhage and resuscitation in rats. AB - OBJECTIVE: We sought to determine whether treatment with lisofylline (LSF) preserves intestinal barrier function in rats subjected to hemorrhagic shock and resuscitation (HS/R). SETTING: Research laboratory at a major university teaching hospital. DESIGN: Rats were bled to a mean arterial pressure of 30 mm Hg and maintained at that pressure for 90 mins. One group (n = 8) was treated with LSF (bolus doses of 15 mg/kg at 45 and 89 min plus infusion at 10 mg x kg(-1) x hr( 1)), whereas another group (n = 8) received only the lactated Ringer's solution (LRS) vehicle. At 90 mins, the animals were resuscitated with shed blood and LRS (55 mL x kg(-1) x hr(-1)). Intestinal mucosal permeability was determined by measuring the mucosal-to-serosal clearance of fluorescein isothiocyanate dextran (molecular weight = 4 kDa) into everted gut sacs. MEASUREMENTS AND MAIN RESULTS: Intestinal and hepatic blood flow (assessed by laser Doppler flowmetry) was greater in LSF-treated rats. Treatment with LSF ameliorated the development of histologic evidence of mucosal damage and hyperpermeability. Rats treated with LSF had lower plasma concentrations of the intracellular hepatic enzyme, aspartate aminotransferase. After 90 mins of resuscitation, concentrations of adenosine triphosphate in intestinal and hepatic tissue were greater in LSF treated as compared with LRS-treated rats, but concentrations of the endogenous antioxidant, glutathione, in intestinal and hepatic tissue, although lower than in rats not subjected to HS/R, were similar in the two treatment groups. CONCLUSION: Treatment with LSF ameliorated HS/R-induced derangements in intestinal structure and function and hepatic injury, possibly by preserving microvascular perfusion and tissue adenosine triphosphate concentrations. PMID- 10834710 TI - Neutrophil depletion in rats reduces burn-injury induced intestinal bacterial translocation. AB - OBJECTIVE: To determine whether neutrophil depletion could eradicate intestinal bacterial translocation in bum-injured rats. DESIGN: Prospective, randomized, controlled study. SETTING: University research laboratory. SUBJECTS: Adult male Sprague-Dawley rats. INTERVENTIONS: The rats were intravenously administered a rabbit anti-rat neutrophil antibody causing profound neutropenia and subjected to a 30% total body surface area scald burn. MEASUREMENTS AND MAIN RESULTS: The depletion of neutrophils from the intestine was assessed via measurements of myeloperoxidase (MPO) activity in the intestinal homogenates. In addition, the presence of activated/extravasated neutrophils in intact intestines was determined via immunohistochemical localization of neutrophil nicotinamide adenine dinucleotide phosphate (NADPH) oxidase component protein p47phox. Bacterial translocation was measured using agar cultures and by determining Escherichia coli beta-galactosidase gene via polymerase chain reaction/Southern blot analyses of mesenteric lymph node and spleen, liver, lung, and blood. MPO measurements demonstrated a six-fold increase above the control value in the intestinal tissue in rats on day 1 postburn. The presence of activated neutrophils (expression of p47phox protein) was also markedly increased in the intestines of these rats. The increased MPO activity and p47phox expression accompanied a translocation of indigenous E. coli into the mesenteric lymph node without a spread to other organs. The administration of anti-neutrophil antibody to burn animals prevented an increase in MPO activity and bacterial translocation. CONCLUSION: These studies indicate that enhanced intestinal bacterial translocation caused by burn injury could be related to the increased infiltration of activated neutrophils into the intestinal tissue after bum. The release of neutrophil products such as superoxide anion may effect intestinal tissue damage leading to bacterial translocation of indigenous E. coli. PMID- 10834711 TI - Long-term outcome after medical reversal of transtentorial herniation in patients with supratentorial mass lesions. AB - OBJECTIVE: To determine the short- and long-term outcomes after successful reversal of transtentorial herniation by medical treatment. Although it has been recognized that aggressive medical management can reverse transtentorial herniation, it is believed that overall outcome in such patients is poor. DESIGN: Prospective cohort study. SETTING: Neurocritical care unit of a university hospital. PATIENTS: A total of 28 consecutive patients who underwent an episode of transtentorial herniation (defined as decrease in level of consciousness accompanied by pupillary dilation) secondary to a supratentorial mass lesion followed by successful reversal. INTERVENTION: Herniation was reversed by using a combination of hyperventilation, mannitol and hypertonic saline. MEASUREMENTS AND MAIN RESULTS: The following outcomes were analyzed: risk of second herniation, radiologic evidence of structural damage or vascular compromise related to herniation on post-herniation computed tomographic scan, in-hospital mortality, and long-term functional outcome using Rankin score and Barthel index. A total of 32 episodes of transtentorial herniations were reversed in 28 patients during a 14-month period. The most common precipitating cause were edema (n = 23) or new/expanding intracerebral hematoma (n = 5). After first reversal of transtentorial herniation in 28 patients, a second herniation episode was observed in 16 patients after a mean interval of 88.2 hrs (range, 23-432 hrs); four were successfully reversed. On follow-up computed tomographic scan, hypodense lesion in midbrain (n = 6), temporal lobe contusion (n = 2), posterior cerebral artery (n = 3), and middle cerebral artery (n = 1) infarction were visualized in a minority of patients. The in-hospital mortality was 60% (n = 15) with brain death being the cause of death in 13 patients; care was withdrawn in eight patients. Second episode of herniation (p = .002) and midbrain involvement during herniation (p = .02) were associated with in-hospital mortality. During a mean follow-up period of 11.4+/-4.2 months, two patients died of cerebral neoplasm and human immunodeficiency virus-related sepsis, respectively. Of the 11 survivors, 7 were functionally independent (Rankin score <3 and Barthel index >60). CONCLUSIONS: Although mortality after transtentorial herniation is high, we found a prominent potential for meaningful recovery with aggressive medical reversal of transtentorial herniation. Our study implies that timely medical intervention for reversing transtentorial herniation can result in preservation of neurologic function. PMID- 10834712 TI - Pulse oximeters' reliability in detecting hypoxemia and bradycardia: comparison between a conventional and two new generation oximeters. AB - OBJECTIVE: Pulse oximeters are increasingly used for patient monitoring; however, they are traditionally very prone to motion artifact. Newly developed instruments have lower false alarm rates. We wanted to know whether this is achieved at the expense of an increased proportion of false negative alarms such as missed or delayed identification of hypoxemia and/or bradycardia. DESIGN: Observational study. SETTING: Neonatal intensive care unit. PATIENTS: A total of 17 unsedated preterm infants (median gestational age at birth, 25 wks; range, 24-30 wks). INTERVENTION: Long-term recordings of transcutaneous partial pressure of oxygen (P(Tc)O2), heart rate, pulse oximeter saturation (SpO2), and pulse rate from a conventional oximeter and two new generation oximeters. MEASUREMENTS: Recordings were analyzed for episodes with P(Tc)O2 <40 torr or with heart rate <80 beats/min for >5 secs. Hypoxemia was considered identified if SpO2 had fallen to <85% within 2 mins of P(Tc)O2 reaching 40 torr, and bradycardia was considered identified if pulse rate had fallen to <80 beats/min within 2 mins of the heart rate reaching this threshold. MAIN RESULTS: A total of 202 falls in P(Tc)O2 to <40 torr occurred; 174 (86%) were identified by all three oximeters. Of the remaining episodes, manual analysis of red and infrared absorption signals confirmed that SpO2 had indeed been <85% for > or =10 secs in 11 episodes; therefore, these episodes should have been identified by all three oximeters. None of these had been missed by the conventional oximeter, but 10 (5.4% of the total) were missed by one of the new generation instruments (Nellcor), and one (0.5%) was missed by the other (Masimo). Of 54 bradycardias, only 14 were identified by all three oximeters; 17 (32%) were missed by the conventional, 37 (69%) by the Nellcor, and 4 (7%) by the Masimo instrument. CONCLUSION: One of the two new generation instruments investigated in this study missed 5.4% of hypoxemic episodes and 69% of bradycardias. It thus appears that this instrument's reduced false alarm rate is achieved at the expense of an unreliable and/or delayed identification of hypoxemia and bradycardia. The other instrument identified both conditions equally as or more reliably than a conventional pulse oximeter. PMID- 10834713 TI - Effect of neuromuscular blockade on oxygen consumption and energy expenditure in sedated, mechanically ventilated children. AB - OBJECTIVE: To quantify the effects of neuromuscular blockade (NMB) on energy expenditure for intubated, mechanically ventilated, critically ill children. DESIGN: A prospective, unblinded clinical study. Each subject was studied twice, before and after establishment of NMB. SETTING: A tertiary care pediatric intensive care unit. PATIENTS: Critically ill children undergoing mechanical ventilation and receiving ongoing sedation were eligible, if they had a cuffed endotracheal tube and were physiologically stable. INTERVENTIONS: A total of 20 children (age, 1 to 15 yrs) were studied in an unblinded, crossover fashion. All were mechanically ventilated via a cuffed endotracheal tube, with ventilator rate and tidal volume adequate to provide complete ventilation, and F(IO2) <0.6. Absence of gas leak around the endotracheal tube was assured, and all patients were sedated using continuous infusions of midazolam and/or fentanyl; no changes in ventilator settings, nutritional input, or inotropic drug dose were permitted during the study period. Each patient underwent indirect calorimetry immediately before establishment of NMB. NMB was then induced, and indirect calorimetry was repeated. Complete blockade was verified using a peripheral nerve stimulator. In each case, the two sets of measurements were completed within a 1-hr period. MEASUREMENTS AND MAIN RESULTS: Data analyzed included identifying and diagnostic information, oxygen consumption, and carbon dioxide production. Energy expenditure was calculated using standard formulas. Oxygen consumption and energy expenditure values obtained before and after the establishment of NMB were compared by using paired Student's t-test. NMB reduced oxygen consumption from 6.54+/-0.49 mL/kg/min to 5.90+/-0.40 ml/kg/min, and energy expenditure was reduced from 46.5+/-3.7 kcal/kg/24 hrs to 41.0+/-2.8 kcal/kg/24 hrs (p < .001 in each case). The reduction in oxygen consumption was 8.7+/-1.7%, and that in energy expenditure 10.3+/-1.8%, of pre-NMB values, respectively. CONCLUSION: NMB significantly reduces oxygen consumption and energy expenditure in critically ill children who are sedated and mechanically ventilated; the degree of reduction is small. PMID- 10834714 TI - Preterm infants with high polyunsaturated fatty acid and plasmalogen content in tracheal aspirates develop bronchopulmonary dysplasia less often. AB - OBJECTIVE: Oxygen toxicity causes chronic bronchopulmonary dysplasia (BPD) in extremely preterm infants. Polyunsaturated fatty acids (PUFA) and plasmalogens are the two main substrates for lipid peroxidation in the pulmonary surfactant. In the present study, we tested whether low concentrations of both were associated with development of BPD and whether both were further reduced during mechanical ventilation with oxygen. DESIGN: Prospective, noninterventional, descriptive study. SETTING: Level III neonatal intensive care unit in a university hospital. PATIENTS: In 25 extremely low birth weight infants with respiratory distress syndrome, tracheal aspirates were collected immediately after birth and in the following 4 days. As control, tracheal and pharyngeal aspirates were collected from healthy infants immediately after birth. The amount of PUFA and dimethylacetals (DMA, representing plasmalogens) was determined gas chromatographically. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The relative percentages of PUFA and DMA on all fatty acids in non-BPD infants (PUFA% 26+/-8.9, DMA% 3.5+/-1.2) were higher compared with infants who developed BPD (PUFA% 14.5+/-3.8, DMA% 1.8+/-0.9). In term healthy infants, DMA% and PUFA% were in the same range as in the BPD group. The higher levels found for non-BPD infants decreased after day 1 to values equal to the BPD group and remained low. CONCLUSIONS: The results suggest that initially higher levels of PUFA and plasmalogens in the tracheal effluent are associated with a reduced risk of developing BPD and are reduced during the first day of ventilation. PMID- 10834716 TI - The value of gastric intramucosal pH in the postoperative period of cardiac surgery in pediatric patients. AB - OBJECTIVE: To examine the usefulness of gastric intramucosal pH (pHi) to predict adverse events during the postoperative period after cardiac surgery in pediatric patients. DESIGN: Prospective, observational clinical study. SETTING: Multidisciplinary pediatric intensive care unit, university hospital. PATIENTS: A total of 70 patients who had elective cardiac surgery and cardiopulmonary bypass; mean age, 4.0 yrs. INTERVENTIONS: Gastric tonometry. MEASUREMENTS AND MAIN RESULTS: In all cases, the pHi was determined within the first 2 hrs and again within the first 12 hrs of the postoperative period after cardiac surgery. The following adverse events were considered: mortality, multiple organ failure (MOF), and infectious complications. Patients were divided into three groups: Group A, patients having the two pHi measurements < or =7.32 (n = 15); Group B, patients having the two pHi determinations >7.32 (n = 28); and Group C, patients having one pHi determination < or =7.32 and the other >7.32 (n = 27). Of the total sample, 5 (7.1%) patients died; 4 (5.7%) had MOF, and 13 (18.5%) had infectious complications. When the rate of adverse events was analyzed in the three groups, significant differences were found in mortality (p < .035) and MOF (p < .029). This was not the case for infectious complications (p = .071). In Group A, three patients died and three had MOF. The standardized chi-square values were 1.87 and 2.31 respectively, causing the authors to consider that this group accounted for the differences found in the global analysis. CONCLUSIONS: Two pHi determinations < or =7.32, one at admission and one at 12 hrs of the postoperative period after cardiac surgery and cardiopulmonary bypass, were associated with a higher rate of mortality and MOF, but not with a higher rate of infectious complications. It is not yet possible to recommend the use of this finding as a treatment objective to guide hemodynamic optimization in this group of patients. PMID- 10834715 TI - Recombinant human growth hormone treatment in pediatric burn patients and its role during the hepatic acute phase response. AB - OBJECTIVE: Recombinant human growth hormone (rHGH) has been shown to increase mortality in adult trauma patients; however, little has been reported on its side effects in children. The acute phase response has been suggested to be a contributing factor to trauma mortality. Therefore, the purpose of this study was to examine the effects of exogenous rHGH on the acute phase response in pediatric bum patients. DESIGN: Prospective, randomized, double-blind study. SETTING: Shriners Hospital for Children. PATIENTS: Thermally injured pediatric patients, ranging in age from 0.1 to 16 yrs. INTERVENTIONS: Twenty-eight thermally injured children received either 0.2 mg/kg/day of rHGH or saline (placebo) within 3 days of admission and for at least 25 days. MEASUREMENTS AND MAIN RESULTS: Measurements were patient demographics, incidence of sepsis, inhalation injury, mortality, serum constitutive proteins, acute phase proteins, proinflammatory cytokines and insulin-like growth factor-I (IGF-I), insulin-like growth factor binding protein (IGFBP)-1, and IGFBP-3. No differences could be demonstrated in age, gender, burn size, incidence in sepsis (20% vs. 26%), inhalation injury (46% vs. 27%), or mortality (8% vs. 7%) between those receiving rHGH or placebo. Serum IGF-I and IGFBP-3 increased with rHGH treatment, whereas serum IGFBP-1 decreased compared with placebo (p < .05). Burned children treated with rHGH required significantly less albumin substitution to maintain normal levels compared with placebo (p < .05). Those receiving rHGH demonstrated a decrease in serum C reactive protein and serum amyloid-A and an increase in serum retinol-binding protein compared with placebo (p < .05). rHGH decreased serum tumor necrosis factor-alpha and interleukin (IL)-1beta, whereas no changes were found for serum IL-1alpha, IL-6, and IL-10 compared with placebo (p < .05). Free fatty acids were elevated in burned children who received rHGH (p < .05). CONCLUSION: Data indicate that rHGH does not increase mortality. rHGH decreased acute phase proteins, tumor necrosis factor-alpha, and IL-1beta, which is associated with increases in constitutive hepatic proteins and IGF-I. PMID- 10834717 TI - Attitudes toward limitation of support in a pediatric intensive care unit. AB - OBJECTIVE: To prospectively determine opinions of members of a pediatric intensive care unit (PICU) team regarding the appropriateness of aggressive care. The types of support that caregivers sought to limit and their reasons for wanting these limits were collected over time. DESIGN: Prospective survey of caregiver opinions. SETTING: PICU in an academic tertiary care children's hospital. SUBJECTS: A total of 68 intensive care nurses, 11 physicians attending in the PICU, 10 critical care and anesthesia fellows, and 24 anesthesia and pediatric residents. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: During a 6-month period, 503 patients were admitted to the PICU. Within this time period, 52.4% of all deaths were preceded by limitation of support, with 100% of noncardiac surgical deaths preceded by limitation of medical interventions. At least one caregiver wished to limit care for 63 of these patients (12.5%). When caregivers wished to limit support they most frequently wished to limit invasive modes of support such as cardiopulmonary resuscitation (94%) and hemodialysis (83%). The ethical rationales identified most often for wishing to limit support were burden vs. benefit (88%) and qualitative futility (83%). Preadmission quality of life was cited less frequently (50%). Caregivers were less likely to limit care on the basis of quality of life. Nurses and physicians in the PICU were very similar to each other in the types of support they thought should be limited and their ethical rationales. CONCLUSIONS: When making decisions about whether or not to limit care for a patient, caregivers were more likely to rely on the perceived benefit to the patient than preadmission quality of life. PMID- 10834718 TI - Resistance of pediatric and neonatal endotracheal tubes: influence of flow rate, size, and shape. AB - OBJECTIVE: The aim of this study was to determine the resistances of endotracheal tubes (ETTs) commonly used in neonatal and pediatric intensive care units and the relationship of resistance to flow rate, size, and shape of ETT. SETTING: Laboratory-based measurements. EVALUATION: We examined straight tubes with inner diameters between 2.5 and 6 mm and shouldered (Cole) tubes with inner diameter/outer diameter between 2.5/4 and 3.5/5 mm. We assessed ETT resistance at standard and "appropriate for patient use" lengths at flow rates from 0 L/min to 30 L/min. We used calibrated rotameters to control the flow of gas and proximal static pressure measured by using either an industrial draft gauge or a differential pressure transducer. The ETT resistance was calculated by dividing the proximal ETT pressure by the measured flow and expressed as the mean of three measurements at each flow rate. MAIN RESULTS: Resistance increased as ETT diameter decreased; at flows of 5 L/min and 10 L/min, the resistances of the 6 mm inner diameter ETT were 3.1 H2O/L/sec and 4.6 cm H20/L/ sec, respectively, and the resistances of the 2.5 mm inner diameter ETT were 81.2 H2O/L/sec and 139.4 cm H20/L/sec, respectively. Shortening an ETT to a length appropriate for patient use (e.g., a 4.0 mm inner diameter, from 20.7 to 11.3 cm) reduced its resistance on average by 22%. The resistance of a Cole tube was approximately 50% lower than that of a straight tube with an inner diameter corresponding to the narrow part of the shouldered tube. CONCLUSIONS: Our results suggest that the use of a small diameter, straight ETT will significantly increase the work of breathing. PMID- 10834719 TI - Evaluation of a new transcardiac conductance method for continuous on-line measurement of left ventricular volume. AB - OBJECTIVE: To evaluate a new, less invasive, conductance method to measure continuous on-line left ventricular volume. End-systolic and end-diastolic volumes obtained with this transcardiac conductance method were compared with simultaneous measurements using the conventional intracardiac conductance catheter. DESIGN: Controlled animal study. SETTING: Research laboratory in a university hospital. SUBJECTS: Six sheep. INTERVENTIONS: Anesthetized sheep were instrumented and inotropic condition was varied by beta-receptor stimulation (5 microg/kg/min of dobutamine) and beta-receptor blockade (1 mg/kg of propranolol). In each condition (control, dobutamine, repeat control, propranolol), ventricular volume was varied over a wide range by gradual preload reduction using a vena caval balloon catheter. MEASUREMENTS AND MAIN RESULTS: We compared the two methods by performing linear regression analysis on simultaneous end-systolic and end-diastolic volumes obtained during gradual caval occlusions. We statistically analyzed the intercepts, slopes, and correlation coefficients of the regression equations relating the transcardiac and conductance catheter measurements to determine the effects of interanimal variability, inotropic condition, and cardiac phase on the relationship between the two methods. The results show an excellent linear correlation between the two methods (mean intercept, -1.82+/ 1.24 mL; mean slope, 0.787+/-0.024 and r2 = .94). Both slope and intercept of the relationship between the two methods show a significant interanimal and cardiac phase related variability but no significant dependence on inotropic condition. CONCLUSIONS: The significant interanimal variability indicates that the new method requires individual calibration in each subject. However, the small variability of the regression coefficients with changes in condition indicates that after initial calibration, end-systolic and end-diastolic volume can be followed accurately even in the presence of large changes in volume and inotropic state. This new method may facilitate quantitative continuous assessment of cardiac function in clinical practice, for example, in the intensive care unit. PMID- 10834720 TI - During neonatal mechanical ventilatory support, the delivered nitric oxide concentration is affected by the ventilatory setting. AB - OBJECTIVE: To assess whether the delivered nitric oxide (NO) concentration is affected by a change in the ventilatory setting during neonatal mechanical ventilatory support. DESIGN: Prospective, experimental study. SETTING: Laboratory at Nagoya City University Medical School. INTERVENTIONS: This study was performed by using a pressure-limited, time-cycled, ventilatory support with a neonatal circuit and a 50-mL silicone test lung. NO in N2 gas was administrated into the inspiratory limb at a distance of 4 cm, 80 cm, or 160 cm from the Y piece connected to the adapter of an endotracheal tube. The NO concentration was measured every 0.5 sec by a chemiluminescence analyzer at the Y piece. MEASUREMENT AND MAIN RESULTS: NO concentrations were compared with each of the ventilatory settings of peak inspiratory pressure (PIP) (10-30 cm H2O), positive end-expiratory pressure (0-10 cm H2O), ventilatory flow (10, 20, 30 L/min), and ventilatory rate (30, 40, 50, 60, 70 breaths/min), respectively. The NO concentration was significantly lower when NO was added at 4 cm than at 80 cm or 160 cm from Y piece at the same ventilatory setting of PIP, positive end expiratory pressure and ventilatory flow, respectively, (p < .01). Although the NO concentration was increased as the settled PIP level was increased (p < .01 or p < .05), it was not changed when the settled positive end-expiratory pressure level was increased. A decrease was seen in the NO concentration as the settled ventilatory flow was increased (p < .01). Lastly, the NO concentration fluctuated greatly in association with the settled ventilatory rate. CONCLUSION: The NO concentration delivered to patients is influenced by the ventilatory setting during neonatal mechanical ventilatory support. PMID- 10834721 TI - Compensation for teaching in critical care. AB - OBJECTIVES: To determine the financial or nonclinical time critical care program directors or teaching faculty members receive as compensation for their educational activities. To compare compensation types and amounts among critical care specialties and between university vs. nonuniversity sponsoring institutions. DATA SOURCES AND EXTRACTION: Survey returns (46%) from critical care fellowship directors listed in the American Medical Association Graduate Medical Education Directory. Information was stratified according to fellowship specialty and type of sponsoring hospital and compared by chi-square analysis and the Kruskal-Wallis test. CONCLUSIONS: Most program directors (77%) and faculty (82%) receive no specified compensation for education-related activities. Multidisciplinary programs are more likely to compensate faculty members than other specialty-specific programs (p = .006). Most programs sponsored by university or military/federal hospitals do not provide specified compensation (79% and 100%, respectively). Overall, community hospital-based programs provide a greater percentage of compensation to directors and faculty than university programs (for directors, p = .02; odds ratio, 3.85; for faculty, p = .001; odds ratio, 8.4). When compensation is specified, it is most often financial and it averages 18% of the salary (range, 5% to 100%) for directors and 19% of the salary for faculty (range, 5% to 50%). When reduced clinical time is provided (5% of program directors, 2% of faculty), it averages 13% (range, 8% to 18%) for directors and 18% (range, 10% to 25%) for faculty. Alternative methods for assigning educational compensation are discussed. PMID- 10834722 TI - Which therapeutic interventions in critical care medicine have been shown to reduce mortality in prospective, randomized, clinical trials? A survey of candidates for the Belgian Board Examination in Intensive Care Medicine. AB - OBJECTIVE: To highlight the lack of randomized controlled trial (RCT) evidence in support of accepted therapeutic interventions in the intensive care unit by assessing the ability of Intensive Care Board Examination candidates to cite relevant studies. DESIGN: As part of the Board Examination for Intensive Care Medicine in Belgium, candidates were asked to name accepted therapeutic interventions that have been shown to reduce mortality in RCTs. SETTING: Survey of doctors. SUBJECTS: Candidates for the 1998 Board Examination for Intensive Care Medicine in Belgium. INTERVENTIONS: None MEASUREMENTS AND MAIN RESULTS: Of the 46 candidates, 25 (54%) gave no response and 3 (7%) stated that no such study exists; 16 responses from 13 doctors were considered acceptable answers. CONCLUSIONS: Candidates for the Board Examination of Intensive Care in Belgium were unable to list many accepted interventions in critical care medicine that are supported by RCT evidence. The RCT may not be the most appropriate tool for assessing the value of an intervention in the intensive care environment. PMID- 10834723 TI - Neurologic manifestations of cerebral air embolism as a complication of central venous catheterization. AB - OBJECTIVE, PATIENTS, AND METHODS: A severe case of cerebral air embolism after unintentional central venous catheter disconnection was the impetus for a systematic literature review (1975-1998) of the clinical features of 26 patients (including our patient) with cerebral air embolism resulting from central venous catheter complications. RESULTS: The jugular vein had been punctured in eight patients and the subclavian vein, in 12 patients. Embolism occurred in four patients during insertion, in 14 patients during unintentional disconnection, and in eight patients after removal and other procedures. The total mortality rate was 23%. Two types of neurologic manifestations may be distinguished: group A (n = 14) presented with encephalopathic features leading to a high mortality rate (36%); and group B (n = 12) presented with focal cerebral lesions resulting in hemiparesis or hemianopia affecting mostly the right hemisphere, with a mortality rate as high as 8%. In 75% of patients, an early computed tomography indicated air bubbles, proving cerebral air embolism. Hyperbaric oxygen therapy was performed in only three patients (12%). A cardiac defect, such as a patent foramen ovale was considered the route of right to left shunting in 6 of 15 patients (40%). More often, a pulmonary shunt was assumed (9 of 15 patients; 60%). For the remainder, data were not available. CONCLUSION: When caring for critically ill patients needing central venous catheterization, nursing staff and physicians should be aware of this potentially lethal complication. PMID- 10834724 TI - Phlegmasia cerulea dolens with compartment syndrome: a complication of femoral vein catheterization. AB - OBJECTIVE: Central venous catheterization is commonly performed in the critically ill. The femoral vein is widely accepted as an insertion site with complications thought to be comparable to other central access sites. We used serial ultrasound examinations with Doppler to examine the evolution of a heretofore undescribed complication of femoral vein catheterization, phlegmasia cerulea dolens with compartment syndrome. DESIGN: Serial ultrasounds were performed in patients before the insertion of femoral venous catheters and sequentially every 48 hrs while the catheters were in place. The noncatheterized leg served as a control. SETTING: A trauma and life support center of a tertiary multidisciplinary critical care unit. PATIENT: A 32-yr-old man with respiratory failure as a consequence of a severe community-acquired pneumonia that required central venous access for antibiotics because no peripheral sites could be obtained. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The initial ultrasound examination of both legs before femoral catheter insertion revealed no sign of venous thrombosis. Ultrasound of the catheterized leg at 48 hrs revealed a small nonocclusive thrombosis, whereas the opposite leg remained normal. At 72 hrs, the catheterized leg had clinical and ultrasonographic evidence of a massive thrombosis. A compartment syndrome defined by pressure measurements soon ensued and required emergent surgical release. CONCLUSIONS: This case report and a review of the available literature suggest that thrombosis associated with femoral vein catheterization should be considered when clinicians decide where to obtain central venous access when multiple sites are available. This report also suggests the utility of serial ultrasound examinations to define clinically nonapparent thrombosis as an early indicator of a potentially catastrophic complication. PMID- 10834725 TI - Hyperlactatemia, increased osmolar gap, and renal dysfunction during continuous lorazepam infusion. AB - OBJECTIVE: To review effects of the vehicle of lorazepam, propylene glycol, in regard to lactate, osmolarity, and renal dysfunction. DESIGN: Case report. SETTING: Intensive care unit of a Level I trauma center. Patient A 36-yr-old Hispanic man who developed severe respiratory failure and required high-dose lorazepam for sedation. The patient was ventilated with low tidal volumes in a lung-protective fashion, with resultant "permissive hypercapnia." Lactates and osmolalities rose on initiation and fell, as expected, on discontinuation of the lorazepam infusion. However, there was no renal compensation for the hypercapnia except while the patient was not receiving lorazepam. MEASUREMENTS AND MAIN RESULT: Serial osmolalities, lactates, serum bicarbonate, PaCO2, and pH were measured during lorazepam infusion. Rise and fall of serum lactate and osmolality closely correlated with lorazepam. Serum bicarbonate rose significantly while the patient was not receiving lorazepam in response to hypercarbia and failed to rise while the patient was receiving lorazepam. CONCLUSION: The vehicle of lorazepam, propylene glycol, can cause hyperlactatemia and elevated osmolar gaps. However, propylene glycol may also interfere with renal tubular function and may blunt renal compensation for respiratory acidosis. PMID- 10834726 TI - Transient swallow syncope during periods of hypoxia in a 67-year-old patient after self-extubation. AB - OBJECTIVE: To describe the case of an adult patient with swallow syncope after bypass surgery, possibly related to hypoxia. DESIGN: Case report. SETTING: University hospital, medical-cardiologic intensive care unit. PATIENT: A 67-yr old patient after second aortocoronary bypass operation for unstable angina. MAIN RESULTS: After the patient managed to extubate himself, he was in a borderline respiratory condition with an oxygen mask. When drinking for the first time after extubation, asystole was observed coincidentally with interruption of oxygen insufflation. During the next days, similar events occurred during food ingestion or when drinking liquids after a fall of oxygen saturation. The bradyarrhythmia was readily reversible on administration of atropine and ventricular backup pacing via temporary pacing wires. After normalization of gas exchange, no more episodes of swallowing-associated asystole were observed and the patient was discharged without a permanent pacemaker. There was no esophageal or gastrointestinal disease. Pre- and postoperative PR and QRS durations were normal. CONCLUSION: Extrinsic and transient mechanisms, rather than intrinsic conduction system disease, seem to have been operative in this case. It is suggested that hypoxia reinforced the vagal pharyngocardiac reflex as described in pediatric patients. PMID- 10834727 TI - Severe transmyocardial ischemia in a patient with tension pneumothorax. AB - OBJECTIVE: To report tension pneumothorax (TP) as a cause of severe myocardial ischemia. DESIGN: Clinical case report. SETTING: Medical intensive care unit of a university hospital. PATIENTS: One patient with severe shock attributable to right TP after unsuccessful percutaneous central venous catheterization. INTERVENTIONS: Blood pressure, electrocardiogram (ECG), chest radiograph, and echocardiography during and after shock. MEASUREMENTS AND MAIN RESULTS: On admission the patient was in profound state of shock (heart rate 140 beats/min, blood pressure 65/30 mm Hg). Twelve-lead ECG showed pronounced ST segment elevation in leads II, III, aVF, and V4-V6. Chest radiograph revealed right TP with complete displacement of the mediastinum and the heart to the left side. Immediate right-sided tube thoracostomy resulted in reexpansion of the lung followed by instantaneous hemodynamic and respiratory improvement as well as nearly complete resolution of the ECG changes. Peak value of the creatine phosphokinase was 4140 U/L without significant elevation of the MB isoenzyme at any time. Moreover, the initial hypokinesia of the posterior and lateral left ventricular wall resolved completely, as demonstrated by echocardiography. CONCLUSION: The specific condition of TP may lead to impaired systolic and diastolic coronary artery blood flow affecting ventricular repolarization and T wave configuration in ECG indicative of transmyocardial ischemia. General symptoms, namely hypotension, tachycardia, and hypoxemia, are likewise typical for cardiogenic shock attributable to myocardial infarction. Yet any therapeutic measure directed toward revascularization, such as thrombolysis or even percutaneous transluminal coronary angioplasty, would have had devastating consequences. Therefore, thorough physical examination of our patient was pivotal in disclosing the true origin of profound shock. PMID- 10834728 TI - Clinical assessment and management of massive hemoptysis. AB - OBJECTIVE: Massive hemoptysis is a potentially lethal condition that deserves to be investigated thoroughly and brought under control promptly. The mortality rate depends mainly on the underlying etiology and the magnitude of bleeding. Although the diagnosis of hemoptysis may be established by chest radiograph, many pathologies may be missed. Because bronchoscopy and computed tomography are complementary, they may indicate pathologies not detectable by chest radiograph. Finding the etiology and site of the hemoptysis is imperative. INVESTIGATIONS: Urgent bronchoscopy should be performed in unstable patients because it exacts a paramount role in the diagnostic search and therapy. It can be used to facilitate the introduction of balloon-tip catheters into the bleeding bronchus for tamponade of the hemorrhagic artery, protecting de facto the contralateral lung or nonbleeding bronchi from blood aspiration. Endobronchial tamponade should only be used as a temporary measure until a more specific treatment is instituted. In stable patients, computed tomography should be ordered before any bronchoscopic exploration. INTERVENTIONS: Surgery was once regarded as the treatment of choice in operable patients with massive hemoptysis. Bronchial artery embolization (BAE) is an excellent nonsurgical alternative; it is proven to be very effective and lacks the mortality and morbidity encountered in surgical interventions. Nevertheless, surgery is recommended in patients with massive hemoptysis caused by thoracic vascular injury, arteriovenous malformation, leaking thoracic aneurysm with bronchial communication, hydatid cyst, and other conditions in which BAE would be inadequate. MEDICAL MANAGEMENT: Conservative medical therapy may suffice in certain conditions, like bronchiectasis, coagulopathies, Goodpasture's syndrome, and acute bronchopulmonary infections. Preparation for other interventions (endobronchial tamponade, BAE, or surgery in eligible candidates) should be undertaken if the bleeding fails to respond to conservative measures. Supportive therapy should be applied vigorously to all patients with massive hemoptysis. PMID- 10834729 TI - Chest physiotherapy in mechanically ventilated children: a review. AB - OBJECTIVE: Many physicians, nurses, and respiratory care practitioners consider chest physiotherapy (CP) a standard therapy in mechanically ventilated children beyond the newborn period. CP includes percussion, vibration, postural drainage, assisted coughing, and suctioning via the endotracheal tube. DATA SOURCES: We searched the medical literature by using the key words "chest physiotherapy" and "chest physical therapy" (among others) by means of the MEDLINE and Current Contents databases. STUDY SELECTION: Because of the paucity of objective data, we examined all reports dealing with this topic, including studies on adult patients. For data extraction, not enough material existed to perform a meta analysis. DATA SYNTHESIS: Despite its widespread use, almost no literature dealing with this treatment modality in pediatric patients exists. Studies with mechanically ventilated pediatric and adult patients have shown that CP is the most irritating routine intensive care procedure to patients. An increase in oxygen consumption often occurs when a patient receives CP accompanied by an elevation in heart rate, blood pressure, and intracranial pressure. CP leads to short-term decreases in oxygen, partial pressure in the blood, and major fluctuations in cardiac output. Changes in these vital signs and other variables may be even more pronounced in pediatric patients because the lung of a child is characterized by a higher closing capacity and the chest walls are characterized by a much higher compliance, thus predisposing the child to the development of atelectasis secondary to percussion and vibration. CONCLUSION: CP in mechanically ventilated children may not be considered a standard therapy. Controlled studies examining the impact of CP on the duration of mechanical ventilatory support, critical illness, and hospital stay are needed. PMID- 10834730 TI - Pressure support for the acute respiratory distress syndrome: less is more. PMID- 10834731 TI - Recumbent position and autonomic function in cardiac patients: will "Right to Sleep" be the next campaign? PMID- 10834732 TI - Opioids, febrile responsiveness, and thinking outside the box. PMID- 10834733 TI - Predicting mortality in intensive care unit patients with stroke. PMID- 10834734 TI - Renal dose dopamine: long on conjecture, short on fact. PMID- 10834735 TI - Pulmonary dysfunction after cardiopulmonary bypass: should we ventilate the lungs on pump? PMID- 10834736 TI - Pain assessment in the seriously ill patient: can family members play a role? PMID- 10834737 TI - A peek at renal blood flow, renal function, and oxygen consumption with epinephrine and dopamine therapy. PMID- 10834738 TI - Tracheostomy: timing is everything. PMID- 10834739 TI - Percutaneous dilational tracheostomy: a note of caution. PMID- 10834740 TI - Alpha-melanocyte stimulating hormone in sepsis: is your patient's brain smarter than your patient's doctor? PMID- 10834741 TI - Prolonged use of heat and moisture exchangers: why do we keep changing things? PMID- 10834742 TI - Is big NO little NO's evil twin? PMID- 10834743 TI - Use of epinephrine as vasopressor agent during cardiopulmonary resuscitation: another example of a double-edged sword. PMID- 10834744 TI - Round and round we go, and where we stop, nobody knows: the open lung concept and high-level positive end-expiratory pressure (revisited) PMID- 10834745 TI - Intratracheal pulmonary ventilation, the latest new ventilation technique for supporting diffuse lung injury: do we jump on the bandwagon? PMID- 10834746 TI - Partial liquid breathing: are we headed in the right direction? PMID- 10834747 TI - The frustrating problem of smoke inhalation injury. PMID- 10834748 TI - Myocardial effects of epinephrine during ventricular fibrillation: does flow matter? PMID- 10834749 TI - Heat shock proteins, oxygen radicals, and apoptosis: the conflict between protection and destruction. PMID- 10834750 TI - Hemofiltration-absorption systems for the treatment of experimental sepsis: is it possible to remove the "evil humors" responsible for septic shock? PMID- 10834751 TI - Is bacterial translocation a clinically relevant phenomenon in burns? PMID- 10834752 TI - Pulse oximetry's final frontier. PMID- 10834753 TI - Deciding whether to withdraw life-support in critically ill children: insightful data on hard choices. PMID- 10834754 TI - Measuring critical heart data in the critical care setting. PMID- 10834755 TI - Faculty reimbursement for graduate medical education: who gives and who receives? PMID- 10834756 TI - Extracorporeal life support in pediatric acute respiratory failure: we can afford it AND need it. PMID- 10834757 TI - BIPAP: useful new modality? PMID- 10834758 TI - Salt water: a solution? PMID- 10834759 TI - Reliability of the anion gap. PMID- 10834760 TI - Lung protective strategies: just going back to basics. PMID- 10834761 TI - Is sodium bicarbonate therapy during cardiopulmonary resuscitation really detrimental? PMID- 10834762 TI - Protein C concentrate and recombinant tissue plasminogen activator in meningococcal septic shock. PMID- 10834763 TI - Is albumin administration harmful in critically ill patients? The patient's nitrogen and energy balance may provide an answer. PMID- 10834764 TI - Physiological and behavioral regulation in two-year-old children with aggressive/destructive behavior problems. AB - A sample of 99 two-year-old children was selected on the basis of parents' responses to two administrations of the Child Behavior Checklist for two- to three-year-olds. Forty-nine of these children displayed symptoms of aggressive/destructive (externalizing) problems that were in the borderline clinical range (labelled "high risk") and 50 children displayed few such symptoms ("low risk"). The children were assessed in a series of laboratory procedures that were intended to be emotionally and behaviorally challenging, during which time heart rate was recorded and behavior was observed. To assess physiological regulation, resting measures of heart period and respiratory sinus arrhythmia (RSA), and heart period change and RSA suppression were derived from these procedures. To assess emotional and behavioral regulation, children's affect and on-task versus types of off-task behaviors were measured. Results indicated that children in the high-risk group did not differ from children in the low-risk group on the resting measure of heart period. Boys displayed lower heart rate than did girls, regardless of risk group. However, boys in the low-risk group differed from boys in the high-risk group in terms of resting measures of RSA. Children in the high-risk group did display significantly and consistently lower RSA suppression (physiological regulation) during the challenging situations than did the children in the low-risk group. High-risk children displayed more negative affect and dysregulated emotion regulation behaviors than did the low risk children. These findings are discussed in terms of the development of behavioral and emotional regulation that underlie adaptive versus maladaptive behavior. PMID- 10834765 TI - Early developmental precursors of externalizing behavior in middle childhood and adolescence. AB - This study examined the infancy- and toddler-age precursors of children's later externalizing problem behavior. Risk constructs included suboptimal patterns of observed caregiver-child interaction and the caregiver's perception of child difficultness and resistance to control. In addition, a novel dimension of caregiver-child relationship quality, the caregiver's perception of her toddler's unresponsiveness to her, was examined as a possible precursor of children's externalizing behavior. Externalizing problem outcomes were assessed throughout the school-age period and again at age 17, using multiple informants. As toddlers, children at risk for later externalizing behavior were perceived as difficult and resistant to control, and relationships with their caregivers were relatively low in warmth and affective enjoyment. Finally, the caregiver's perception of her toddler as emotionally unresponsive to her was a consistent predictor of later externalizing behavior, suggesting that negative maternal cognitions associated with child conduct problems may begin in toddlerhood. These predictive patterns were similar for boys and girls, and with minor exceptions, generalized across different subdimensions of externalizing problem behavior. Our findings underscore the importance of the infancy and toddler periods to children's long-term behavioral adjustment, and indicate the desirability of further research into the nature of caregivers' early perceptions of child unresponsiveness. PMID- 10834766 TI - The efficacy of toddler-parent psychotherapy for fostering cognitive development in offspring of depressed mothers. AB - The efficacy of Toddler-Parent Psychotherapy (TPP) as a preventive intervention for fostering cognitive development in the offspring of depressed mothers was evaluated. Mothers with major depressive disorder and their toddlers were randomly assigned to TPP (n = 43) or to a nonintervention group (n = 54) and compared to a control group (n = 61) of women with no current or past mental disorder. At baseline (age 20 months), the groups did not differ on the Bayley Mental Development Index. At post-intervention follow-up (age 3 years), a relative decline in IQ was found in the depressed nonintervention group, whereas the depressed intervention and the normal control groups continued to be equivalent, with higher WPPSI-R Full Scale and Verbal IQs. The worst outcome was found among nonintervention children whose mothers had subsequent depressive episodes. The results confirm the developmental risks faced by offspring of depressed mothers and support the efficacy of the preventive intervention in safeguarding successful cognitive development in at-risk youngsters. PMID- 10834767 TI - Etiology of inattention and hyperactivity/impulsivity in a community sample of twins with learning difficulties. AB - A community sample of 373 8 to 18 year-old twin pairs in which at least one twin in each pair exhibited a history of learning difficulties was utilized to examine the etiology of inattention and hyperactivity/impulsivity (hyp/imp). Symptoms of attention-deficit/hyperactivity disorder (ADHD) were assessed by the DSM-III Diagnostic Interview for Children and Adolescents. Inattention and hyp/imp composite scores were created based on results of a factor analysis. Results indicated that extreme ADHD scores were almost entirely attributable to genetic influences across several increasingly extreme diagnostic cutoff scores. Extreme inattention scores were also highly heritable whether or not the proband exhibited extreme hyp/imp. In contrast, the heritability of extreme hyp/imp increased as a linear function of the number of inattention symptoms exhibited by the proband. This finding suggests that extreme hyp/imp may be attributable to different etiological influences in individuals with and without extreme inattention. If this result can be replicated in other samples, it would provide evidence that the hyp/imp symptoms exhibited by individuals with Combined Type ADHD and Predominantly Hyp/Imp Type ADHD may be attributable to different etiological influences. PMID- 10834768 TI - A cross-domain growth analysis: externalizing and internalizing behaviors during 8 years of childhood. AB - In a sample of 405 children assessed in kindergarten through the seventh grade, we determined the basic developmental trajectories of mother-reported and teacher reported externalizing and internalizing behaviors using cross-domain latent growth modeling techniques. We also investigated the effects of race, socioeconomic level, gender, and sociometric peer-rejection status in kindergarten on these trajectories. The results indicated that, on average, the development of these behaviors was different depending upon the source of the data. We found evidence of the codevelopment of externalizing and internalizing behaviors within and across reporters. In addition, we found that African American children had lower levels of externalizing behavior in kindergarten as reported by mothers than did European-American children but they had greater increases in these behaviors when reported by teachers. Children from homes with lower SES levels had higher initial levels of externalizing behaviors and teacher reported internalizing behaviors. Males showed greater increases in teacher reported externalizing behavior over time than did the females. Rejected children had trajectories of mother-reported externalizing and internalizing behavior that began at higher levels and either remained stable or increased more rapidly than did the trajectories for non-rejected children which decreased over time. PMID- 10834769 TI - Subtypes of victims and aggressors in children's peer groups. AB - This study reports an investigation of the behavioral profiles and psychosocial adjustment of subgroups of victims and aggressors in elementary school peer groups. Peer nomination scores for aggression and victimization were used to classify 354 inner-city children (mean age of 10.3 years) into one of four subgroups: aggressive victims, nonaggressive victims, nonvictimized aggressors, and normative contrasts. Subgroup comparisons were then conducted using multi informant assessment of social behavior, social acceptance-rejection, behavioral regulation, academic functioning, and emotional distress. Children in each of the victim-aggressor subgroups were characterized by a degree of social and behavioral maladjustment. However, impairments in behavioral and emotional regulation were most evident for the aggressive victim subgroup. Aggressive victims were also characterized by academic failure, peer rejection, and emotional distress. The results of this investigation highlight the distinctive nature of the aggressive victim subgroup. PMID- 10834770 TI - "The frog ate the bug and made his mouth sad": narrative competence in children with autism. AB - This study compares the narrative abilities of 13 children with autism, 13 children with developmental delays, and 13 typically developing children matched on language ability. Although groups did not differ in their use of causal language or internal state terms, children with autism and children with developmental delays were less likely than typical children to identify the causes of characters' internal states. Rather, they tended simply to label emotions and explain actions. Children with autism and children with developmental delays also relied on a more restricted range of evaluative devices, which both convey point of view and maintain listener involvement. In addition, the narrative abilities of children with autism were linked to performance on measures of theory of mind and an index of conversational competence, whereas this was not the case among children with developmental delays. Findings are discussed in relation to the social, cognitive, and emotional underpinnings and consequences of narrative activity. PMID- 10834771 TI - Dysfunctional cognitions in children with social phobia, separation anxiety disorder, and generalized anxiety disorder. AB - According to cognitive theories of anxiety, anxious adults interpret ambiguous situations in a negative way: They overestimate danger and underestimate their abilities to cope with danger. The present study investigated whether children with social phobia, separation anxiety disorder, and generalized anxiety disorder have such a bias, compared to a clinical and a normal control group. Children were exposed to stories in which ambiguous situations were described, and asked to give their interpretations, using open and closed responses. Results showed that anxious children reported more negative cognitions than control children. However, anxious children did not overestimate danger on the free responses, but they did judge the situations as more dangerous on the closed responses. Anxious children had lower estimations of their own competency to cope with danger than the control groups on both open and closed responses. The results indicate that children with anxiety disorders have dysfunctional cognitions about ambiguous situations. PMID- 10834772 TI - To avoid "transient neurologic symptoms"--the search continues. PMID- 10834773 TI - Procaine compared with lidocaine for incidence of transient neurologic symptoms. AB - BACKGROUND AND OBJECTIVES: Transient neurologic symptoms (TNS) have been reported to occur after 16% to 40% of ambulatory lidocaine spinal anesthetics. Patient discomfort and the possibility of underlying lidocaine neurotoxicity have prompted a search for alternative local anesthetic agents. We compared the incidence of TNS with procaine or lidocaine spinal anesthesia in a 2:1 dose ratio. METHODS: Seventy outpatients undergoing knee arthroscopy were blindly randomized to receive either 100 mg hyperbaric procaine or 50 mg hyperbaric lidocaine. An interview by a blinded investigator established the presence or absence of TNS, defined as pain in the buttocks or lower extremities beginning within 24 hours of surgery. Onset of sensory and motor block, patient discomfort, supplemental anesthetics, and side effects were recorded by the unblinded managing anesthesia team. Anesthetic adequacy was determined from these data by a single blinded investigator. Hospital discharge time was recorded from the patient record. Groups were compared using appropriate statistics with a P < .05 considered significant. RESULTS: TNS occurred in 6% of procaine patients versus 31% of lidocaine patients (P = .007). Sensory block with procaine and lidocaine was similar, while motor block was decreased with procaine (P < .05). A trend toward a higher rate of block inadequacy (17% v 3%, P = .11) and intraoperative nausea (17% v 3%, P = .11) occurred with procaine. Average hospital discharge time with procaine was increased by 29 minutes (P < .05). CONCLUSIONS: The incidence of TNS was substantially lower with procaine than with lidocaine. However, procaine resulted in a lower overall quality of anesthesia and a prolonged average discharge time. If the shortfalls of procaine as studied can be overcome, it may provide a suitable alternative to lidocaine for outpatient spinal anesthesia to minimize the risk of TNS. PMID- 10834774 TI - Effects of immediately initiating an epidural infusion in the combined spinal and epidural technique in nulliparous parturients. AB - BACKGROUND AND OBJECTIVES: Intrathecal fentanyl with bupivacaine provides rapid labor analgesia of limited duration. We investigated the effect of initiating an epidural infusion of 0.1% ropivacaine with fentanyl 2 microg/mL and epinephrine 1:400,000 (REF) on the duration of analgesia and incidence of side effects after intrathecal injection in the combined spinal and epidural technique. METHODS: Thirty-four nulliparous parturients with a cervical dilation of 3 to 5 cm were randomized to receive epidural saline or REF at 10 mL following the intrathecal injection of fentanyl 25 microg and bupivacaine 2.5 mg. Degree of analgesia, severity of pruritus, motor block, blood pressure, and sensory level to coolness were assessed until the patient requested additional analgesia. RESULTS: Analgesia was significantly longer in the REF group, 158.4 +/- 59.6 minutes versus 103.8 +/- 26.2 minutes. The decrease in blood pressure compared with the blood pressure at intrathecal injection was greater for the REF group at all times, but achieved statistical significance at 60 minutes. There was no difference in ephedrine use, pruritus, or motor block between groups. There was no difference in sensory level to coolness at 90 minutes after intrathecal injection between groups. CONCLUSIONS: Initiating an infusion of REF prolongs the duration of analgesia, but also results in a greater decrease in blood pressure. Despite this effect on blood pressure, there was no difference in ephedrine use. PMID- 10834775 TI - Effect of epidural epinephrine infusion with bupivacaine on labor pain and mother fetus outcome in humans. AB - BACKGROUND AND OBJECTIVES: Epinephrine is used with local anesthetics to prolong the duration of epidural analgesia and decrease the peak plasma concentrations of local anesthetics. In the practice of obstetric anesthesia, the utero-placental and fetal effects of epinephrine are controversial issues. We designed a prospective, randomized, and double-blind study to examine the effects of epinephrine infusion on the quality of analgesia and uterine or umbilical blood flows with Doppler ultrasound, as well as the duration of the first or the second stage of labor, and fetal outcome. METHODS: Twenty-eight parturients received continuous epidural bupivacaine 0.25% (4 mL/h) combined either with epinephrine (20 microg/h) (n = 13) or without epinephrine (n = 15) for analgesia during labor. If patients requested additional analgesia, an additional bolus of 1% or 1.5% lidocaine (6 to 10 mL) was administered. RESULTS: The total amount of additional lidocaine was greater in the plain bupivacaine group (130 [0, 280] mg; median [25th, 75th percentile] with P < .05) than in the epinephrine group (0 [0, 60] mg). Epinephrine infusion did not alter the resistance of the uterine and umbilical arteries as measured by resistance index. The duration of the first or second stages of labor did not significantly differ in the 2 groups. Epinephrine infusion did not change the fetal heart rate or the blood gas data in the umbilical artery. CONCLUSIONS: A low-dose epidural infusion of epinephrine decreased anesthetic requirements. PMID- 10834776 TI - Low-dose bupivacaine-fentanyl spinal anesthesia for cesarean delivery. AB - BACKGROUND AND OBJECTIVES: The hypotension following spinal anesthesia remains commonplace in cesarean delivery. Intrathecal opioids are synergistic with local anesthetics and intensify sensory block without increasing sympathetic block. The combination makes it possible to achieve spinal anesthesia with otherwise inadequate doses of local anesthetic. We hypothesized that this phenomenon could be used to provide spinal anesthesia for cesarean delivery while incurring less frequent hypotension. METHODS: Thirty-two women scheduled for cesarean delivery were divided into 2 groups of patients who received a spinal injection of either 10 mg of isobaric (plain) bupivacaine 0.5% or 5 mg of isobaric bupivacaine with 25 microg fentanyl added. Each measurement of a systolic blood pressure less than 95 mm Hg or a decrease in systolic pressure of greater than 25% from baseline was considered as hypotension and treated with a bolus of 5 to 10 mg of intravenous ephedrine. RESULTS: Spinal block provided surgical anesthesia in all patients. Peak sensory level was higher (T3 v T4. 5) and motor block more intense in the plain bupivacaine group. The plain bupivacaine patients were more likely to require treatment for hypotension (94% v 31%) and had more persistent hypotension (4.8 v 0.6 hypotensive measurements per patient) than patients in the minidose bupivacaine-fentanyl group. Mean ephedrine requirements were 23.8 mg and 2.8 mg, respectively, for the 2 groups. Patients in the plain bupivacaine group also complained of nausea more frequently than patients in the minidose bupivacaine fentanyl group (69% v 31%). CONCLUSIONS: Bupivacaine 5 mg + fentanyl 25 microg provided spinal anesthesia for cesarean delivery with less hypotension, vasopressor requirements, and nausea than spinal anesthesia with 10 mg bupivacaine. PMID- 10834777 TI - Bupivacaine-sparing effect of fentanyl in spinal anesthesia for cesarean delivery. AB - BACKGROUND AND OBJECTIVES: Visceral pain decreases in cesarean patients under spinal anesthesia when the dose of local anesthetic is increased. However, larger doses of local anesthetic are associated with higher sensory blocks. We hypothesized that the addition of fentanyl could reduce the dose of bupivacaine necessary to achieve adequate surgical anesthesia. METHODS: Two double-blinded, sequential, prospective studies were performed on 120 patients. In the preliminary study, the patients received 8, 10, or 12 mg of 0.5% hyperbaric bupivacaine intrathecally. In the second, main study, they received each bupivacaine dose with 10 microg of fentanyl. Each group consisted of 20 patients, and the groups were identified as B8, B10, B12, BF8, BF10, and BF12. Sensory and motor block, intraoperative pain defined by visual analogue scale (VAS), muscle relaxation, and side effects were assessed. We also measured the sensory and motor recovery and the onset of pain in the postanesthesia care unit (PACU). RESULTS: Maximal block level and incidence of high block (> or = T1) were higher in the 12-mg groups. There was intraoperative pain in 35% of the B8 patients and 20% of the B10 patients, but none in the B12 patients and all fentanyl groups. Incidences of other side effects were not different. The addition of fentanyl to bupivacaine significantly delayed the onset of postoperative pain and sensory recovery, but motor recovery time did not change with additional fentanyl. CONCLUSIONS: The optimal dose of hyperbaric bupivacaine to produce surgical anesthesia was 12 mg, which was accompanied by high sensory block. With the addition of 10 microg of fentanyl, the dose of bupivacaine could be reduced to 8 mg in spinal anesthesia for cesarean delivery. PMID- 10834778 TI - A randomized comparison of the effects of continuous thoracic epidural analgesia and intravenous patient-controlled analgesia after posterior spinal fusion in adolescents. AB - BACKGROUND AND OBJECTIVES: Pain and gastrointestinal dysfunction are primary factors that delay recovery after posterior spinal fusion. Previous reports suggest that the choice of analgesic management may effect the course of recovery. This prospective, randomized study compared continuous thoracic epidural analgesia and patient-controlled analgesia in the postoperative care of adolescents undergoing posterior spinal fusion for idiopathic scoliosis. METHODS: Patients between 11 and 18 years of age were randomized to receive continuous thoracic epidural analgesia with bupivacaine-fentanyl (CEA, n = 17) or intravenous patient-controlled analgesia with morphine sulfate (n = 16). After surgery under general anesthesia, pain intensity was evaluated using a self report visual analog scale (VAS). Postoperative time to resumption of bowel sounds, liquid intake, and side effects were also recorded. RESULTS: There were no significant differences between groups in VAS pain scores, side effects, or time to resumption of liquid intake. There was a significant difference (P = .0089) between groups in return of bowel sounds, which occurred earlier in patients receiving CEA. CONCLUSIONS: Continuous epidural analgesia and patient controlled analgesia are comparably effective and safe after posterior spinal fusion. Return of bowel sounds occurred significantly more rapidly in patients receiving CEA postoperatively. PMID- 10834779 TI - Patient-controlled epidural analgesia with fentanyl-bupivacaine: influence of prior dural puncture. AB - BACKGROUND AND OBJECTIVES: Combined spinal epidural anesthesia (CSEA) involves the epidural administration of local anesthetic and opioid solutions adjacent to the prior dural puncture, potentially increasing their diffusion into the subarachnoid space. This study was designed to evaluate the influence of dural puncture on the adequacy and extent of analgesia, and drugs requirements of patient-controlled epidural analgesia (PCEA) in the postoperative period. METHODS: In this prospective double-blind study, 40 patients undergoing major abdominal surgery under general anesthesia followed with PCEA were randomly assigned to either group I (preoperative insertion of an epidural catheter) or group II (preoperative dural puncture with a 25-g Quincke needle + insertion of an epidural catheter). Postoperatively, a PCEA pump delivered an infusion of 0.1% bupivacaine + fentanyl (3 microg/mL) at 5 mL/h. Participants were allowed to self administer 5-mL boluses of the same solution with a 15-minute lock-out interval. Hourly epidural solution requirements were recorded for 40 hours. Sensory and motor block, and pain scores were also analyzed. RESULTS: There was no difference between groups with regard to epidural solution requirements, pain scores, spread of sensory blockade, or intensity of motor block. CONCLUSION: Dural puncture with a 25-gauge Quincke needle, performed as part of CSEA, does not influence the drug requirements when a combination of 0.1% bupivacaine and fentanyl (3 microg/mL) is used for PCEA after major abdominal surgery. PMID- 10834780 TI - The minimum effective concentration of opioids: a revisitation with patient controlled analgesia fentanyl. AB - BACKGROUND AND OBJECTIVES: Whether patients titrate themselves to an individualized blood or plasma opioid concentration (the so-called minimum effective concentration or [MEC]) has been debated extensively. Nevertheless, there is consistent opinion that during patient controlled analgesia (PCA) patients balance acceptable pain relief against unacceptable side effects. This study sought to characterize fentanyl used by PCA with respect to MEC and factors influencing PCA use. METHODS: An intensive study of 25 patients with observations over the first 24 hours after orthopedic surgery was planned on the premise that this approach would provide a measure of the fentanyl MEC. This necessitated repeated measurements of pain scores and plasma fentanyl concentrations before and 10 minutes after every PCA demand. In addition, a battery of psychological tests was given before and approximately 48 hours after surgery. RESULTS: Logistic difficulties of maintaining a 24-hour study design resulted in its termination after 5 patients. The patients had convincingly distinct MECs (ranging from 0.23 to 0.99 ng/mL). The relationship between plasma fentanyl concentration and pain score was steep, such that small changes in concentration coincided with marked changes in pain relief. Despite preoperative expectations of achieving satisfaction in postoperative analgesia, not all patients titrated themselves to a pain-free state; all but one were satisfied with PCA. Surprisingly few side effects were reported. Unfortunately, the small sample size made systematic analysis of the psychological tests impossible. CONCLUSIONS: This study found evidence to support the concepts of an individual MEC and a therapeutic window of fentanyl used with PCA. PMID- 10834781 TI - Plasma lidocaine, monoethylglycinexylidide, and glycinexylidide concentrations after epidural administration in geriatric patients. AB - BACKGROUND AND OBJECTIVES: The purpose of this study was to evaluate the effect of age on the pharmacokinetics of lidocaine after epidural administration. METHODS: Two percent lidocaine with epinephrine (5 microg/mL) was administered in two different age groups: an adult group (age 42 +/- 6 years, n = 10) and an elderly group (age 77 +/- 4 years, n = 10). Concentrations of lidocaine and its active metabolites, monoethylglycinexylidide (MEGX) and glycinexylidide (GX), were measured in plasma samples obtained after 15, 30, 45, 60, 90, 120, 150, and 180 minutes of administration using high-performance liquid chromatography with ultraviolet detection. RESULTS: No significant differences in plasma concentrations of lidocaine and its metabolites were observed between the two groups during the 3 hours of study. However, the elderly group showed significantly longer mean residence times (MRTs) and lower plasma clearance of lidocaine during the period compared with the adult group (P < .05). Plasma concentration ratios of MEGX/lidocaine were significantly lower in the elderly group after 2 hours of lidocaine administration (P < .05). CONCLUSIONS: The increase in plasma lidocaine concentration after epidural anesthesia in elderly patients was not as high as anticipated. However, the elderly patients showed longer MRTs, lower clearance, and lower ratios of MEGX/lidocaine than did the adult (middle-age) patients. PMID- 10834782 TI - Ziconotide, a new N-type calcium channel blocker, administered intrathecally for acute postoperative pain. AB - BACKGROUND AND OBJECTIVES: Voltage-sensitive calcium channel conductance is essential for the nervous system to signal a painful event. However, intrathecal administration of L-type calcium channel blockers does not provide analgesia. The present investigation was designed to assess the safety and analgesic efficacy of ziconotide, a new N-type calcium channel blocker, when administered intrathecally to patients with acute postoperative pain. METHODS: This randomized, double blind, pilot study included patients undergoing elective total abdominal hysterectomy, radical prostatectomy, or total hip replacement. After intrathecal injection of local anesthetic and before surgical incision, a continuous intrathecal infusion of either placebo or 1 of 2 doses of ziconotide (0.7 microg/h or 7.0 microg/h) was started and continued for 48 to 72 hours postoperatively. Primary and secondary efficacy variables were the mean daily patient controlled analgesia (PCA) morphine equivalent consumption and visual analog pain intensity (VASPI) scores, respectively. RESULTS: Thirty patients received study drug; 26 were evaluable for efficacy. Mean daily PCA morphine equivalent consumption was less in patients receiving ziconotide than in placebo treated patients, and the difference was statistically significant between 24 and 48 hours (P = .040). VASPI scores during the first 8 hours postoperatively were markedly lower in ziconotide-treated than in placebo-treated patients. In 4 of 6 patients receiving the high-dose of ziconotide (7 microg/h), adverse events, such as dizziness, blurred vision, nystagmus, and sedation contributed to study drug being discontinued after 24 hours. After ziconotide discontinuation, these symptoms resolved. CONCLUSIONS: Ziconotide showed analgesic activity, as shown by decreased PCA morphine equivalent consumption and lower VASPI scores. Because of a favorable trend of decreased morphine consumption with an acceptable side effect profile in the low-dose ziconotide group, 0.7 microg/h may be closer to the ideal dose than 7 microg/h. Large-scale studies are required to clarify this issue. PMID- 10834783 TI - Effect of high-dose lidocaine treatment on superoxide dismutase and malon dialdehyde levels in seven diabetic patients. AB - BACKGROUND AND OBJECTIVES: We report on the use of intravenous (IV) high-dose lidocaine to relieve diabetic neuropathic pain, and the technique's effects on clinical measures of lipid peroxidation. METHODS: Under continuous electrocardiogram monitoring, IV lidocaine (5 mg kg(-1) in 100 mL saline) was administered over 30 minutes to 7 non-insulin-dependent diabetic patients suffering from neuropathic pain who reported increased pain within the preceding 6 months. This treatment was performed once a week for 1 month. Blood samples were collected from the contralateral limb to determine plasma superoxide dismutase (SOD) and malondialdehyde (MDA) levels on admission and following the final lidocaine administration. RESULTS: Plasma MDA concentrations significantly decreased after the final IV lidocaine treatment (P < .05, paired t-test), whereas SOD levels did not show a statistically significant difference compared with baseline levels. CONCLUSIONS: High-dose lidocaine treatment lessens MDA levels, a marker of free-radical-mediated cell damage. This suggests that one of lidocaine's mechanism of action may be its effect on oxygen free radicals, which in turn impacts lipid peroxidation. PMID- 10834784 TI - Neuropathic pain: role of voltage-dependent calcium channels. PMID- 10834786 TI - On-line resources for the Pain Medicine physician. AB - BACKGROUND AND OBJECTIVES: The Internet is increasingly an important resource for both the Pain Medicine physician and the patient. Considerable high-quality information exists within numerous government, university, professional association, and private sites. Large documents, such as patient care guidelines, can be readily retrieved on-line. Search capabilities within websites improve the efficiency of information retrieval. Clinicians may communicate about complex patient management issues via a list-serve, which delivers entire discussions to their E-mail accounts. These multiple resources offer an enriched environment for patient care, education, and research in pain management. PMID- 10834785 TI - Radiographic appearance of the lumbar spine after lumbar fusion. PMID- 10834787 TI - Georg Perthes--the man behind the technique of nerve-tracer technology. PMID- 10834788 TI - Myofascial pain in patients with postthoracotomy pain syndrome. AB - OBJECTIVE: Postthoracotomy pain syndrome is generally considered to be neuropathic pain due to intercostal nerve injury. However, nonneuropathic pain can also occur following thoracic surgery. We present a series of cases with postthoracotomy pain syndrome in which myofascial pain was thought to be a causative component of postthoracotomy pain syndrome. CASE REPORT: Twenty-seven patients (17 men and 10 women) were treated with trigger point injections, intercostal nerve blocks, and/or epidural blocks. Clinical criteria were used to diagnose the myofascial pain. A visual analogue scale was used, and sensory disturbances were recorded before and after treatment. A trigger point in a taut muscular band within the scapular region, which we diagnosed as myofascial pain, was observed in 67% of the patients. The existence of this trigger point significantly increased the rate of success for the treatments. CONCLUSIONS: Postthoracotomy pain may result, at least in part, from a nonneuropathic origin (myofascial pain). It is recommended that each patient be examined in detail to determine whether there is a trigger point in a taut muscular band within the scapular region. If found, this point is suggested as a good area for anesthetic injection. PMID- 10834789 TI - Determining epidural catheter location using nerve stimulation with radiological confirmation. AB - BACKGROUND AND OBJECTIVES: The use of epidural stimulation to confirm epidural catheter placement has been shown. This case report describes the benefits and problems of using the epidural stimulation test to confirm epidural catheter placement and provides supporting evidence for these observations using radiological imaging. CASE REPORT: METHODS: A nerve stimulator was connected to the proximal end of an epidural catheter via an adapter. The cathode lead was connected to the adapter. The anode lead was connected to an electrode placed on the upper extremity as a grounding site. Using 1 to 10 mA current, a segmental motor response indicated that the catheter was in the epidural space. The absence of a motor response indicated that it was not. CASES: In the first patient, the new test predicted subcutaneous epidural catheter placement, which was subsequently confirmed radiologically. In the second patient, the catheter tip was found to be lying near a nerve root, which was again confirmed radiologically. In the third case, a negative test was initially observed with only local muscle movement over the biceps area (T2). After relocation of the grounding electrode to the lower extremity, segmental intercostal muscle movement (T4-5 level) was observed. The catheter placement was radiologically shown to be in the T4-5 region. CONCLUSION: This report illustrates some of the potential benefits and problems of using the nerve stimulation test to confirm epidural catheter placement, with radiological verification. PMID- 10834790 TI - Calciphylaxis treated with neurolytic lumbar sympathetic block: case report and review of the literature. AB - BACKGROUND AND OBJECTIVE: Calciphylaxis is a painful complication of end-stage renal disease and secondary hyperparathyroidism. Calcification most commonly affects skin and soft tissue of the lower extremities resulting in excruciatingly painful skin ulcers. Treatment involves correction of hypercalcemia and hyperphosphatemia, parathyroidectomy, and supportive measures. METHODS: The literature and the merits of neurolytic lumbar sympathetic blockade (LSB) for the treatment of pain associated with calciphylaxis are reviewed. CONCLUSIONS: The neurolytic LSB provided pain relief and is a treatment modality to be considered in managing the pain associated with calciphylaxis. PMID- 10834791 TI - Lateral cervical epidural catheter placement for continuous unilateral upper extremity analgesia and sympathetic block. AB - BACKGROUND AND OBJECTIVES: The use of the laterally directed cervical epidural catheter for the treatment of acute and chronic pain has not been previously described. We have used this technique in a series of 30 patients to produce unilateral upper extremity analgesia and sympathetic block. We present a case report of a patient treated with this technique and a description of the technique and results from the series. METHODS: We placed an epidural catheter at the C6-7 vertebral interspace and directed the needle bevel and catheter to the affected side in a woman with a history of complex regional pain syndrome of the right upper extremity. RESULTS: We were able to show a unilateral block with this technique by anatomic parameters (unilateral contrast dye spread), physiologic parameters (unilateral Horner's syndrome), and symptomatic parameters (unilateral, subjective neural block). Although 1 patient treated with this technique had bilateral effects, no patient has had a predominant block on the contralateral side and there have been no complications. CONCLUSIONS: This case report shows results typical of this series of 30 patients. In this series, the laterally directed cervical epidural catheter was an effective technique to produce continuous unilateral analgesia and sympathetic block. Key Words: Analgesia (epidural), Autonomic nerve block, Reflex sympathetic dystrophy, Postoperative pain. PMID- 10834792 TI - Complete brachial plexus palsy after total shoulder arthroplasty done with interscalene block anesthesia. AB - BACKGROUND AND OBJECTIVES: This report illustrates that brachial plexus palsy can result from either interscalene block or total shoulder arthroplasty. It is often impossible to determine which procedure caused the deficit; therefore, we believe the focus should be placed on treatment of the neurologic deficit. This report provides a suggested algorithm for diagnosis and treatment of postprocedure brachial plexus palsy. METHODS: Interscalene block was used as the operative anesthetic for our patient's total shoulder arthroplasty. Complete brachial plexus palsy was diagnosed postoperatively. RESULTS: The patient's postoperative treatment and recovery are described. CONCLUSIONS: Proper diagnosis and treatment of postprocedure brachial plexus palsy may improve recovery of function. Several precautions may reduce the likelihood of brachial plexus palsy following interscalene block for total shoulder arthroplasty. PMID- 10834793 TI - Management of hypotension during spinal anesthesia. PMID- 10834794 TI - Statistically significant? PMID- 10834795 TI - Lumbar spine pain originating from vertebral osteophytes. PMID- 10834796 TI - Use of intrathecal hyperbaric methadone in postoperative analgesia of thoracic surgery. PMID- 10834797 TI - Variability of a thoracic paravertebral block. Are we ignoring the endothoracic fascia? PMID- 10834798 TI - Fluoroscopic imaging during supraclavicular lateral paravascular brachial plexus block. PMID- 10834799 TI - Treatment of pain from vertebral compression fractures caused by osteoporosis. PMID- 10834800 TI - Time and incidence of paresthesia comparing spinocath with microintralong. PMID- 10834801 TI - Health partnerships for public health development: a New Zealand perspective. PMID- 10834802 TI - Public health strategies for Maori. AB - When the New Zealand Department of Public Health was established in 1900, Maori health status was compromised to the extent that survival itself was threatened. The remarkable turnaround was unexpected and owes much to pioneer Maori professionals, especially the first Maori medical practitioner, Dr. Maui Pomare, who graduated in the United States in 1899. As "Medical Officer to the Maoris," and later as Minister of Health, he made major changes through a five-part strategy: recognizing Maori community leaders as leaders in health, improving access to societal goods and services (especially housing and education), appealing to cultural practices that were linked to good health, wise use of political power, and developing a skilled Maori health workforce to complement community leadership. Although mental health disorders and lifestyle illnesses have largely replaced infectious diseases, malnutrition, and tuberculosis, Pomare's strategy has continuing relevance and warrants serious consideration as a model for health promotion. PMID- 10834803 TI - A moment in time: selected results from the 1996-1997 New Zealand health survey. AB - This article presents key results from the 1996-1997 New Zealand Health Survey, the second of two nationally representative surveys on the health status and health service utilization of New Zealanders. This survey involved a face-to-face interview with 7,862 adults and 1,019 children carried out during the period of a year. The survey had an adult response rate of 73.8%. Using the results of this survey, this article highlights some of the major public health issues facing New Zealanders. These issues include smoking, physical inactivity, alcohol use, asthma, and diabetes. The results presented here show that significant disparities exist between particular demographic and socioeconomic groups in New Zealand in relation to both health risk factors and specific chronic diseases. The policy implications of the results are discussed in relation to existing public health strategies and future initiatives. PMID- 10834804 TI - Why do Australians live longer than New Zealanders? AB - The aim of this study was to compare patterns of mortality in Australia and New Zealand, using routinely collected data. Life expectancy at birth is greater in Australia than in New Zealand (in 1996 the gap was 1.5 years for women and 1.1 years for men). Prior to 1970, mortality was lower in New Zealand than Australia. Possible reasons for the divergence in life expectancies include slower economic growth in New Zealand, more marked increases in economic inequalities which have affected Maori in particular and, to a modest extent, differentials in health care. PMID- 10834805 TI - Socioeconomic deprivation and ethnicity are both important for anti-tobacco health promotion. AB - The aim of the study was to explore the relative importance of socioeconomic deprivation and ethnicity for smoking in New Zealand in order to assist with the design and evaluation of health promotion programs. Smoking data were derived from the 1996 census. Socioeconomic deprivation was measured using the NZDep96 index of socioeconomic deprivation for small areas, which combines nine variables from the 1996 census. There was a strong and consistent relationship between area level socioeconomic deprivation and the proportion of regular smokers. In all age groups, at each level of deprivation, Maori smoked more than the "European and Other" ethnic group. The findings of this study support the view that effective tobacco control activities should address ethnic differences in smoking behavior as well as socioeconomic deprivation, and must operate at the levels of populations, places and environments, as well as individuals. PMID- 10834806 TI - Toward totally smokefree schools and beyond: the Crown Public Health Smokefree Schools Grant Program. AB - A three-year small grants scheme in Canterbury and the West Coast, New Zealand aims to influence the health promoting climate of schools by adopting totally smokefree policies. The Smokefree Environments Act of 1990 established the legal imperative for school policies. The principles of the Treaty of Waitangi and the strategies of the Ottawa Charter provide the programmatic and evaluation framework. Evaluation indicators include the organizational and policy context, status of school smoking policies, reasons for participation, stakeholder involvement, the value of additional funding, and the extent of media coverage. Partnerships among local smokefree partners, schools and whanau/communities, and between the health and education sectors are a priority. Interim findings indicate the Crown Public Health Smokefree Schools Grant program has positively influenced policy development toward totally smokefree schools. Based on these results and the changing environment, the smokefree efforts are being integrated into the broader context of health promoting schools. PMID- 10834807 TI - Te Whanau Cadillac--a waka for change. AB - Community action research is an effective mechanism to improve the relationship between research and public health practice. The Alcohol and Public Health Research Unit (APHRU) and its Maori research partner Whariki have developed practice in building partnerships with communities characterized by mutual respect of the different resources the partners bring to public health. A current APHRU/Whariki project focused on youth and drugs is a partnership with six communities in rural and urban areas throughout New Zealand, many predominantly Maori. This project illustrates a model that aims to bridge the gap between research and practice, and to improve health and well-being through working with communities to increase their capacity to deal with alcohol and drug issues, and to introduce sustainable initiatives. PMID- 10834808 TI - Open houses and closed rooms: Tokelau housing in New Zealand. AB - The migrant Tokelau community in New Zealand, through the Wellington Tokelau Association, is playing an active role in partnership with the Wellington School of Medicine to develop a research program to help improve the health of its people. The relationship between crowded homes and health was investigated in six focus groups. While cultural patterns were an essential part of Tokelau hospitality, the decision to "double up" households was often the result of "rational" economic decision making in relation to household expenditures such as rent and food. The implication for public health practitioners is that while overcrowding may be a health hazard for residents, the most effective solutions by the community are higher household income and more flexible housing designs that accommodate multifamily households. PMID- 10834809 TI - Taking public health to the streets: the 1998 Auckland Meningococcal Disease Awareness Program. AB - Since 1991, New Zealand has experienced a dramatic increase in cases of serogroup B meningococcal disease. Early in 1998, a program was initiated in Auckland to raise awareness among high-risk communities about the early clinical features of meningococcal disease and appropriate action to take. The campaign was conducted largely through door-to-door visiting by Maori and Pacific Islands lay educators, who visited more than 11,000 households and engaged more than 9,000 people in discussion. Significantly greater awareness was found in people visited by lay educators than those not visited. For the first time in 5 years, there were no meningococcal deaths among the target population for the 6 months from the start of the campaign. Home visiting by lay educators is an effective and highly acceptable means of communicating important health information to populations at high risk of meningococcal disease in New Zealand and should be considered for other public health campaigns. PMID- 10834810 TI - Participatory environmental health research in Aotearoa/New Zealand: constraints and opportunities. AB - Within the Aotearoa/New Zealand context, this article identifies opportunities for, as well as constraints on, using participatory research to address environmental health concerns. In New Zealand, principles of partnership fundamental to the Treaty of Waitangi, and the requirements for consultation within the Resource Management Act, provide a framework for participatory approaches. Participatory environmental health research integrates knowledge from various scientific and community sources. It also fosters the innovation, evaluation, and sharing of information that new public health approaches demand. As an emerging field in Aotearoa/New Zealand, it must draw on experience from resource management, rural development, and public health research and practice. Water quality is used as an example of the complexity of environmental health issues and of the potential benefits of engaging stakeholders where long-term health and well-being require balancing ecosystem integrity, economic viability, and social processes. PMID- 10834811 TI - Bacterial resistance and overgrowth due to selective decontamination of the digestive tract. AB - Infection of the lower airways is a major problem in ventilated patients and contributes substantially to morbidity and mortality in the intensive care unit. The selective decontamination of the digestive tract and its effect on the reduction of the gram-negative colonisation rate in patients has been studied widely. However, the findings are inconsistent. Most studies describe an increase in resistant gram-negative bacterial strains and/or an increase in the occurrence of gram-positive strains following selective decontamination of the digestive tract. In light of the unresolved questions concerning the efficacy of selective decontamination of the digestive tract, it would seem that the resultant effect of this treatment on the bacterial flora should be an important consideration when assessing the value of such treatment. To date, none of the studies available for examination have been designed to adequately assess the effect of selective decontamination of the digestive tract on the bacterial flora. PMID- 10834812 TI - Clinical and microbiological survey of Serratia marcescens infection during HIV disease. AB - Clinical charts of 2,398 consecutive HIV-infected patients hospitalized over an 8 year period were reviewed retrospectively to identify all cases of Serratia infection and to evaluate the occurrence and outcome of these cases according to several epidemiological. clinical, and laboratory parameters. Seventeen of 2,398 (0.71%) patients developed Serratia marcescens infections: nine had septicaemia, six had pneumonia, one had a lymph node abscess, and one had cellulitis. All patients were severely immunocompromised, as evidenced by a mean CD4+ lymphocyte count of < 70 cells/microl and a frequent diagnosis of AIDS (13 patients). When compared with other disease localizations, septicaemia was related to a significantly lower CD4+ cell count and a more frequent occurrence of neutropaenia. Antibiotic, corticosteroid, or cotrimoxazole treatment was frequently carried out during the month preceding disease onset. Hospital acquired Serratia spp. infection was more frequent than community-acquired infection and was significantly related to AIDS, neutropaenia, and sepsis. Antimicrobial sensitivity testing showed complete resistance to ampicillin and cephalothin but elevated susceptibility to ureidopenicillins, second- and third generation cephalosporins, aminoglycosides, quinolones, and cotrimoxazole. An appropriate antimicrobial treatment attained clinical and microbiological cure in all cases, in absence of related mortality or relapses. Since only 13 episodes of HIV-associated Serratia spp. infection have been described until now in nine different reports (7 patients with pneumonia, 3 with sepsis, 1 with endophthalmitis, 1 with perifolliculitis, and 1 with cholecystitis), our series represents the largest one dealing with Serratia marcescens infection during HIV disease. Serratia marcescens may be responsible for appreciable morbidity among patients with HIV disease, especially when a low CD4 + cell count, neutropaenia, and hospitalization are present. The clinician and the microbiologist facing a severely immunocompromised HIV-infected patient with a suspected bacterial disease should consider the Serratia spp. organisms. In fact, a rapid diagnosis and an adequate and timely treatment can avoid disease relapses and mortality. PMID- 10834813 TI - Low virulence of Escherichia coli strains causing urinary tract infection in renal disease patients. AB - The distribution of urinary bacterial species was determined and the virulence factors of Escherichia coli urinary strains analysed by molecular and phenotyping methods in episodes of urinary tract infection in renal disease patients (n =68) in comparison with other immunocompromised patients (n =59) and non immunocompromised patients (n =21). Escherichia coli was isolated in 116 (78%) of the 148 patients, being the species most frequently isolated in all groups (75% of renal disease patients, 76% of other immunocompromised patients, 95% of non immunocompromised patients). All other pathogens showed a similar distribution in the renal disease and other immunocompromised patient groups. All virulence factors of Escherichia coli tested for (genes for G adhesins, expression of MR adhesins, production of haemolysin, presence of certain O and K antigens) were found more often in non-immunocompromised than in immunocompromised patients. The factors allowing the highest degree of discrimination between immunocompromised and non-immunocompromised patients were the prevalence of genes for G adhesins (35% vs. 65%) and expression of MR adhesins (32% vs. 55%). It is concluded that there is a lower prevalence of G adhesins and MR adhesins in Escherichia coli strains from immunocompromised patients than non-immunocompromised patients, suggesting that less virulent Escherichia coli strains may cause urinary tract infections more frequently in renal disease patients and other immunocompromised patients. Moreover, the spectrum of urinary pathogens other than Escherichia coli is similar in both immunocompromised patient groups investigated. PMID- 10834814 TI - Measurement of serum antigen concentration by ultrasound-enhanced immunoassay and correlation with clinical outcome in meningococcal disease. AB - The distribution of Neisseria meningitidis serogroup B and C polysaccharide antigen in blood and the prognostic significance of antigen concentration was examined by ultrasound-enhanced immunoagglutination of coated microparticles. Specimens (169 sera/plasma from 145 patients with confirmed meningococcal disease) were tested retrospectively. The ultrasonic immunoassay detected serum antigen in 136 samples from 112 patients. Titration of antigen-positive specimens allowed estimation of blood antigen concentration. The modal blood antigen titre was 1/16, corresponding to an estimated polysaccharide concentration of 0.85 microg/ml. The lowest mean blood antigen concentration found ultrasonically was 0.05 microg/ml; compared to the 1.98 microg/ml found by conventional latex agglutination, this represents an approximately 30-fold improvement in sensitivity. Three grades of outcome were correlated with the presenting antigen titre in 83 patients: (i) <2 weeks hospitalisation, (ii) > or =2 weeks hospitalisation and (iii) mortality. High polysaccharide concentrations correlated with mortality. Nine of 15 patients with a serum antigen titre of 1/64 or greater (> or =3.4 microg/ml polysaccharide) died, whereas no patient with titres equal to or less than 1/4 (< or = 0.21 microg/ml) died, including those patients in whom antigen was undetectable by ultrasonic immunoassay. Increasing antigen concentration significantly correlated with severity of outcome (P<0.001). Ultrasound-enhanced agglutination provides a rapid prognostic indicator by sensitive measurement of serum antigen level. PMID- 10834815 TI - Transient transmission of hepatitis C virus from mothers to newborns. AB - A study was performed in Israel to determine the rate of mother-to-infant HCV transmission in newborns at risk. A group of 22 HCV-infected mothers and their 23 newborns were followed up from early after birth by testing their serum for the presence of HCV antibodies and HCV-RNA. Antibody against HCV was detected in the blood of all newborns immediately after birth, but dropped to low or undetectable levels by 7 months of age. HCV-RNA was detected 2 days after birth in the blood of five infants (22%) but fell to undetectable levels by 6 months. HCV-HVR1 sequence analysis performed in one mother-infant pair on the second day after birth revealed two nucleotide changes. Two months later the same sequence was detected again in the HVR1, suggesting a very low replication rate. Thus, the study showed that vertically transmitted HCV was eliminated in all newborn infants by 6 months after delivery, with concomitant disappearance of HCV antibodies. The mechanism of HCV elimination in newborns at risk remains to be elucidated. PMID- 10834816 TI - Evaluation of phenotypic and genotypic markers for characterisation of the emerging gastroenteritis pathogen Salmonella hadar. AB - Over the last 4 years Salmonella hadar has increasingly been isolated in Europe in conjunction with food-borne gastroenteritis. The aim of this study was to evaluate epidemiological methods (phage typing, antimicrobial susceptibility testing, DNA plasmid analysis, ribotyping and pulsed-field gel electrophoresis analysis) for characterising Salmonella hadar isolates. The 100% phage typeability of isolates and the high discriminatory index of 0.8856 suggest that phage typing is the method of choice. In order to obtain subdivisions of the most frequent Salmonella hadar phage types, a combination of molecular methods, such as ribotyping performed with Bg/I and EcoRI or pulsed-field gel electrophoresis using XbaI and XhoI, would be desirable as the usefulness of each technique varies with the phage type being analysed. Of note was the high (86%) rate of resistance to tetracycline and nalidixic acid but full susceptibility to ciprofloxacin in the strains studied. PMID- 10834817 TI - Prospective surveillance of vancomycin-resistant enterococci in a neonatal intensive care unit. AB - A point-prevalence study of vancomycin-resistant enterococci colonization of the gastrointestinal tract in an Israeli hospital revealed that 14.7% of the 320 inpatients were colonized. Vancomycin-resistant enterococci colonization was detected in most departments except the neonatal intensive care unit. Hence, a prospective longitudinal study of the prevalence of vancomycin-resistant enterococci colonization in the neonatal intensive care unit was conducted. A rectal swab was obtained from every newborn on admission to the neonatal intensive care unit and once weekly thereafter until the patient was discharged. Enterococci were isolated and tested for susceptibility to vancomycin. A total of 84 neonates were enrolled and monitored on average for 3 weeks (SD +/- 3.9, range 1-20 weeks). Mean gestational age was 35.7 weeks (SD +/- 3.9, range 25-42 weeks), and mean birth weight was 2.4 kg (SD +/- 0.9, range 0.45-4.1 kg). Most patients had one or more of the known risk factors associated with colonization with vancomycin-resistant enterococci. Eighty percent of the patients received antibiotics during the study, and 14.3% received vancomycin. The median duration of vancomycin treatment was 12.5 days (SD +/- 16.8, range 5-55 days). Fifty-one of 84 (61%) patients acquired enterococci sensitive to vancomycin during the study period, but no newborn had vancomycin-resistant enterococci. Possible explanations for this finding may be physical isolation of the neonatal intensive care unit from the rest of the hospital, intrinsic differences in the bowel milieu of this age group and the lack of exposure to food and other environmental sources of vancomycin-resistant enterococci from the community. PMID- 10834818 TI - Carriage of antibiotic-resistant Streptococcus pneumoniae in Greek infants and toddlers. AB - The prevalence, resistance patterns and serotypes of antibiotic-resistant Streptococcus pneumoniae strains recovered from Greek carriers under 24 months of age were studied. From February 1997 to April 1998, nasopharyngeal cultures were performed in 1,269 children (ages 2-23 months, median 11 months) living in various areas of central and southern Greece. Resistance (including both intermediate and resistant isolates) to one or more antimicrobial agents was found in 132 of the 421 (31%) Streptococcus pneumoniae isolates, as follows: penicillin, 9% intermediate, 7.6% resistant; cefotaxime, 5.2% intermediate, 0.5% resistant; erythromycin, 0.7% intermediate, 18.1% resistant; clindamycin, 0.2% intermediate, 12.4% resistant; tetracycline, 0.7% intermediate, 16.4% resistant; chloramphenicol, 12.4% resistant; and trimethoprim-sulfamethoxazole, 3.8% intermediate, 14.3% resistant. The MICs of penicillin for 66% of the penicillin nonsusceptible pneumococci were 1-4 microg/ml. Multidrug resistance was found in 64% of penicillin-nonsusceptible and 37% of penicillin-susceptible strains. Sixty two percent of the penicillin-susceptible, multidrug-resistant strains belonged to serotype 6B and were resistant to all five non-beta-lactam agents tested. This notable serotype 6B resistance pattern was described for the first time in a previous study performed from December 1995 to February 1996 in the city of Patras, southwestern Greece. Seventy-two percent of antibiotic-resistant isolates belonged to serotypes 6B, 9V, 14, 18C, 19F and 23F. These results document the spread of resistant pneumococcal strains in central and southern Greece, many of which are multidrug resistant. PMID- 10834819 TI - Cellulitis complicating lymphoedema. AB - In ten hospitalised patients with cellulitis complicating lymphoedema encountered over a 3-year period (1996-1998), the underlying diseases were carcinoma of the cervix (n = 4), uterus (n = 1), vagina (n = 1), breast (n=2) and nasopharynx (n= 1), and retroperitoneal squamous cell carcinoma (n = 1). Three of the ten patients had positive blood cultures, compared to none of the 20 age-matched, sex matched controls hospitalised for cellulitis without lymphoedema. The mean duration of fever, tachycardia and cellulitis was significantly longer in patients with lymphoedema than in those without (P<0.05, P<0.05, and P<0.005 respectively). Early treatment initiated by patients themselves may help stop bacterial replication in the initial stages and minimise further damage to the lymphatic system. PMID- 10834820 TI - Infection with Helicobacter pylori and leukocyte response in patients with myocardial infarction. AB - To test whether Helicobacter pylori may contribute to the inflammatory response following myocardial infarction, the levels of IgG antibodies to Helicobacter pylori and some parameters of leukocyte activity were measured in 63 patients and 61 comparable controls. Helicobacter pylori-positive patients showed a significantly higher expression of the adhesion molecule LFA-1 on neutrophils than Helicobacter pylori-negative patients (433+/-29.0 vs. 398.8+/-38.9 mean fluorescence channels; P<0.0001), whereas no significant difference for any parameters tested was found in control subjects. These data suggest a role of Helicobacter pylori in inducing a leukocyte response following myocardial infarction. PMID- 10834821 TI - Prevalence of Haemophilus influenzae carriers in the Catalan preschool population. Working Group on Invasive Disease Caused by Haemophilus influenzae. AB - This study was designed to determine the prevalence of healthy Haemophilus influenzae carriers in a random sample of the preschool population in Catalonia. Oropharyngeal swabs were collected and cultured on chocolate agar supplemented with 260 microg/ml of bacitracin. Four hundred two of the 734 (54.8%) children studied were detected as Haemophilus influenzae carriers: 7 (0.9%) carried serotype b, 14 (1.9%) serotype e, 6 (0.8%) serotype f, and 375 (51%) carried nontypable strains. The results show that, although the prevalence of Haemophilus influenzae carriers is similar to figures reported from other countries, the prevalence of Haemophillus influenzae serotype b carriers is lower and corresponds with the low incidence of invasive disease observed in the Catalan community. PMID- 10834822 TI - Isolation and continuous growth of Chlamydia pneumoniae from arterectomy specimens. AB - For the purpose of collecting Chlamydia pneumoniae strains of vascular origin that could be grown continuously in vitro, a cell culture system has been established. Using different types of vascular specimens obtained from 38 patients, Chlamydia pneumoniae could be isolated in three (7.9%) cases. The strains were obtained from specimens of the carotid artery, the femoral artery and an infrarenal aneurysm of the abdominal aorta of three male atherosclerosis patients. Thus, viable Chlamydia pneumoniae strains are also present in vascular regions other than those hitherto described. PMID- 10834823 TI - Prevalence of hepatitis E antibodies in immigrants from developing countries. AB - The aim of the present study was to investigate the prevalence of anti-hepatitis E virus (HEV) antibodies among indigenous Spanish blood donors and immigrants from developing countries in order to determine whether immigrants pose a significant risk for the transmission of HEV to the healthy Spanish population. The seroprevalence of HEV was determined in a cohort of 90 asymptomatic immigrants (mostly from countries in sub-Saharan Africa) who had recently arrived in Madrid, Spain, and in 863 blood donors, who represented the healthy Spanish population. The results showed that the prevalence of HEV antibodies was 1.9 times higher in the immigrants than in the blood donors (5.5% in immigrants, 95% CI 1.8-12.4; 2.9% in blood donors, 95% CI 1.9-4.2). Combined with the estimated population figures of 300,000 undocumented immigrants versus 39,000,000 Spaniards, these results indicate that sub-Saharan immigrants cannot currently be considered a major risk source for the transmission of HEV in Spain. PMID- 10834824 TI - Role of bacterial virulence factors and host factors in the outcome of Escherichia coli bacteraemia. AB - In a study of the role of virulence factors in the outcome of Escherichia coli bacteraemia, blood isolates from 30 hospitalised patients were characterised with regard to O and K antigens, P and type 1 fimbriae, haemolysin production, cytonecrotising factor 1 production, serum resistance, ability to activate neutrophils and resistance to killing. Patients were analysed to identify host factors contributing to morbidity and mortality. In univariate analyses the presence of a K antigen, type 1 fimbriae, absence of haemolysin production, serum resistance and resistance to killing were associated with morbidity and mortality. In multivariate analyses only the absence of haemolysin production was associated with morbidity and mortality, after taking host factors into account. These preliminary findings suggest that host factors override bacterial virulence factors in determining the course of Escherichia coli bacteraemia. The negative association between haemolysin production and clinical deterioration during Escherichia coli bacteraemia might indicate predominance of less virulent strains in patients with other risk factors for morbidity and mortality or inactivation of neutrophil products needed for host defence. PMID- 10834825 TI - In vitro susceptibility of environmental Cryptococcus neoformans variety neoformans isolates from Turkey to six antifungal agents, including SCH56592 and voriconazole. AB - The in vitro antifungal susceptibility of 27 environmental (pigeon droppings) isolates of Cryptococcus neoformans var. neoformans, isolated from throughout Turkey, to six antifungal agents (amphotericin B, flucytosine, fluconazole, voriconazole, itraconazole, and SCH56592) was studied. Voriconazole, itraconazole, and SCH56592 all showed comparable activity and were more active than the remaining three antifungal agents tested. Overall, SCH56592 was the most active agent (MIC90, 0.015 microg/ml, at both 48 and 72 h), followed by itraconazole (MIC90, 0.03 microg/ml, at both 48 and 72 h) and voriconazole (MIC90, 0.25 microg/ml, at both 48 and 72 h), respectively. Antifungal susceptibility data for environmental isolates may reflect patterns for the clinical isolates recovered from patients from the same geographic area. PMID- 10834826 TI - Bacteraemia caused by non-glucose-fermenting gram-negative bacilli and Aeromonas species in patients with haematological malignancies and solid tumours. AB - The clinical characteristics and outcome of bacteraemia caused by non-glucose fermenting gram-negative bacilli and Aeromonas spp. were examined in 115 adults with haematological malignancies or solid tumours. The most aggressive pathogens were Pseudomonas aeruginosa, Stenotrophomonas maltophilia, Aeromonas spp., Acinetobacter spp. and Burkholderia cepacia, all of which caused either septic syndrome or pneumonia in more than 40% of cases. Pseudomonas aeruginosa was involved less often in catheter-related bacteraemia than other species. Polymicrobial bacteraemia (n=28) was more often catheter-related than monomicrobial bacteraemia and more often required catheter removal for definitive cure. The most important predictors of catheter-related bacteraemia and its outcome were polymicrobial infection, the presence of pneumonia or septic syndrome and the species involved. PMID- 10834827 TI - Recommendation for prenatal screening for congenital toxoplasmosis. PMID- 10834828 TI - Prevalence and nature of dysphagia in VA patients with COPD referred for videofluoroscopic swallow examination. AB - This retrospective study investigated the prevalence and nature of dysphagia in 78 male outpatients whose primary diagnosis was chronic obstructive pulmonary disease (COPD) and who were referred for modified barium swallow studies at the Veterans Affairs Medical Center (VAMC) in Tucson, Arizona, 1992-1995. Nearly 85% of these patients evidenced some degree of dysphagia, and laryngeal penetrations or aspirations were observed in 44 of them. Agreement between VAMC swallow assessments and those of a three-judge panel of certified speech-language pathologists supported the reliability and validity of the information extracted from VAMC records. The high percentage of silent laryngeal penetrations and aspirations observed in this sample suggests that COPD patient's respiratory status should be considered as well as overt symptoms of a swallowing disorder, such as coughing, in making referrals for video-fluoroscopic swallow studies for such patients. PMID- 10834830 TI - Phonology and children with specific language impairment: status of structural constraints in two languages. AB - Two studies are reported in which the phonological characteristics of preschool age children with specific language impairment (SLI) are compared with those seen in younger normally developing children matched for mean length of utterance and consonant inventory size. The productions of both English-speaking and Italian speaking children with SLI were more likely to deviate from the adult standard than the productions of the younger control children. In Italian, the children with SLI had more difficulty than the younger controls in the use of non-final weak syllables; in English, the children with SLI were poorer than the younger controls in the use of non-final weak syllables, word-final consonants, and word final consonant clusters. These are the same phonological details that are required for several grammatical inflections and many function words in the two languages. However, the children with SLI were also less consistent than their younger compatriots in using consonants in structurally simple words. These findings provide evidence for the view that for many preschool-age children with SLI, phonological problems go beyond those that might be predicted on the basis of the children's short utterances and limited consonant inventories. PMID- 10834829 TI - Distortion product otoacoustic emissions in normal-hearing children with homozygous sickle cell disease. AB - The purpose of this study was to investigate distortion product otoacoustic emissions (DPOAEs) in young normal-hearing children with sickle cell disease (SCD). It was hypothesized that the prevalence of DPOAEs and response amplitudes would be lower than those in children with normal hemoglobin due to suspected compromised cochlear function as a result of vaso-occlusive events characteristic of SCD. Twenty African-American children with SCD and 15 African-American children with normal hemoglobin participated. Distortion product OAEs were evoked by 13 primary tone pairs with f2 frequencies ranging from 1000 to 4500 Hz. The primary tones were presented at L1 and L2 levels of 70 and 60 dB SPL (high) and 50 and 40 dB SPL (low), respectively. The findings of this study were completely unexpected and contrary to our original hypotheses. The likelihood of detecting a DPOAE response was not related to the clinical status of the children. Distortion product OAE amplitudes were significantly larger for children with SCD (p =.01). PMID- 10834831 TI - Diphthong changes in style shifting from Southern English to Standard American English. AB - This study was undertaken to identify the acoustic correlates of the diphthongs /aI/ and /contains as a subset I/ in individuals who were trained to "style shift" between Standard American English (SAE) and Southern English (SE). The diphthongs were produced by four individuals from the coastal southern dialect region who were selected from among those who had successfully completed an accent-reduction program. Results showed that diphthongs used in SE were shorter and had less noticeable transitional elements than those in SAE. The difference was more apparent for /aI/ than /contains as a subset I/. There were no instances in which a diphthong was completely replaced by a vowel in SE. Findings indicate that acoustic analysis may be a useful tool in identifying dialectal elements, particularly for those individuals who are learning to style shift. PMID- 10834832 TI - Acoustic and perceptual characteristics of esophageal and tracheoesophageal speech production. AB - This study comprehensively compared the speech of laryngeal speakers (L), tracheoesophageal speakers (TE), good esophageal speakers (GE), and moderate esophageal speakers (ME) to determine the consequences of TE versus E speech rehabilitation. Twenty speakers (five in each group) were each recorded while reading 16 sentences, and their recordings were analyzed acoustically and perceptually. Acoustic analysis included duration, intensity, fundamental frequency (F0), intonation, and voice onset time measurements. Perceptual analysis included intelligibility and acceptability judgments by naive listeners. The main acoustic results showed that L speakers differ significantly from all alaryngeal speakers in F0 and intonation production. Moderate esophageal speakers differed significantly from all other groups in duration measures. Perceptual results revealed that L speakers were most intelligible and acceptable, whereas ME speakers were least so. Tracheoesophageal speakers were more acceptable than GE speakers but not more intelligible. Significant correlations emerged between F0, duration measures, and acceptability, and between F0 and intelligibility. Also, a significant correlation emerged between acceptability and intelligibility. Findings emphasized the importance of categorizing esophageal speakers into groups based on their speech proficiency level. PMID- 10834833 TI - How soft is a protein? A protein dynamics force constant measured by neutron scattering. AB - An effective environmental force constant is introduced to quantify the molecular resilience (or its opposite, "softness") of a protein structure and relate it to biological function and activity. Specific resilience-function relations were found in neutron-scattering experiments on purple membranes containing bacteriorhodopsin, the light-activated proton pump of halobacteria; the connection between resilience and stability is illustrated by a study of myoglobin in different environments. Important advantages of the neutron method are that it can characterize the dynamics of any type of biological sample-which need not be crystalline or monodisperse-and that it enables researchers to focus on the dynamics of specific parts of a complex structure with deuterium labeling. PMID- 10834834 TI - Distinct roles of CONSTANS target genes in reproductive development of Arabidopsis. AB - In plants, flowering is triggered by endogenous and environmental signals. CONSTANS (CO) promotes flowering of Arabidopsis in response to day length. Four early target genes of CO were identified using a steroid-inducible version of the protein. Two of these genes, SUPPRESSOR OF OVEREXPRESSION OF CO 1 (SOC1) and FLOWERING LOCUS T (FT), are required for CO to promote flowering; the others are involved in proline or ethylene biosynthesis. The SOC1 and FT genes are also regulated by a second flowering-time pathway that acts independently of CO. Thus, early target genes of CO define common components of distinct flowering-time pathways. PMID- 10834835 TI - Gone with the wind: the origin of S0 galaxies in clusters AB - We present three-dimensional, high-resolution hydrodynamical simulations of the interaction between the hot ionized intracluster medium and the cold interstellar medium of spiral galaxies. Ram pressure and turbulent/viscous stripping remove 100% of the atomic hydrogen content of luminous galaxies like the Milky Way within 100 million years. These mechanisms naturally account for the morphology of S0 galaxies and the rapid truncation of star formation implied by spectroscopic observations, as well as a host of observational data on the neutral hydrogen (HI) morphology of galaxies in clusters. PMID- 10834836 TI - Photon-mediated hybridization of frenkel excitons in organic semiconductor microcavities AB - Coherent excitations of intricate assemblies of molecules play an important role in natural photosynthesis. Microcavities are wavelength-dimension artificial structures in which excitations can be made to couple through their mutual interactions with confined photon modes. Results for microcavities containing two spatially separated cyanine dyes are presented here, where simultaneous strong coupling of the excitations of the individual dyes to a single cavity mode leads to new eigenmodes, described as admixtures of all three states. These "hybrid" exciton-photon structures are of potential interest as model systems in which to study energy capture, storage, and transfer among coherently coupled molecular excitations. PMID- 10834837 TI - Light-driven motion of liquids on a photoresponsive surface AB - The macroscopic motion of liquids on a flat solid surface was manipulated reversibly by photoirradiation of a photoisomerizable monolayer covering the surface. When a liquid droplet several millimeters in diameter was placed on a substrate surface modified with a calix[4]resorcinarene derivative having photochromic azobenzene units, asymmetrical photoirradiation caused a gradient in surface free energy due to the photoisomerization of surface azobenzenes, leading to the directional motion of the droplet. The direction and velocity of the motion were tunable by varying the direction and steepness of the gradient in light intensity. The light-driven motion of a fluid substance in a surface modified glass tube suggests potential applicability to microscale chemical process systems. PMID- 10834838 TI - Raman spectroscopy of iron to 152 gigapascals: implications for Earth's inner core AB - Raman spectra of hexagonal close-packed iron (varepsilon-Fe) have been measured from 15 to 152 gigapascals by using diamond-anvil cells with ultrapure synthetic diamond anvils. The results give a Gruneisen parameter gamma(0) = 1.68 (+/-0.20) and q = 0.7 (+/-0.5). Phenomenological modeling shows that the Raman-active mode can be approximately correlated with an acoustic phonon and thus provides direct information about the high-pressure elastic properties of iron, which have been controversial. In particular, the C(44) elastic modulus is found to be lower than previous determinations. This leads to changes of about 35% at core pressures for shear wave anisotropies. PMID- 10834839 TI - Accretion of primitive planetesimals: Hf-W isotopic evidence from enstatite chondrites AB - Enstatite chondrites have often been considered to be closely related to the material from which Earth accreted. However, tungsten isotopic data reveal clear differences. Moreover, the silicate and metal fractions define distinct initial (182)Hf/(180)Hf corresponding to a 13.8 +/- 5.3 million year apparent age difference. Internal reequilibration does not provide a ready explanation for this result. Larger scale redistribution of tungsten is more likely, such as may have occurred during collisions between planetesimals. PMID- 10834840 TI - A monoclinic post-stishovite polymorph of silica in the shergotty meteorite AB - A post-stishovite phase of silica was identified in the Shergotty meteorite by x ray diffraction and field emission scanning electron microscopy. The diffraction pattern revealed a monoclinic lattice, similar to the baddeleyite-structured polymorph with the cell parameters a = 4.375(1) angstroms, b = 4.584(1) angstroms, c = 4. 708(1) angstroms, beta= 99.97(3), rho = 4.30(2) grams per cubic centimeter, where the numbers in parentheses are the maximum deviations. Transmission electron microscopy investigations indicate the presence of the alpha-lead dioxide-like polymorph, stishovite, and secondary cristobalite in the same silica grain. The mixture of high-density polymorphs suggests that several post-stishovite phases were formed during the shock event on the Shergotty parent body. PMID- 10834841 TI - The genetic program of hematopoietic stem cells. AB - Blood cell production originates from a rare population of multipotent, self renewing stem cells. A genome-wide gene expression analysis was performed in order to define regulatory pathways in stem cells as well as their global genetic program. Subtracted complementary DNA libraries from highly purified murine fetal liver stem cells were analyzed with bioinformatic and array hybridization strategies. A large percentage of the several thousand gene products that have been characterized correspond to previously undescribed molecules with properties suggestive of regulatory functions. The complete data, available in a biological process-oriented database, represent the molecular phenotype of the hematopoietic stem cell. PMID- 10834842 TI - Role of 4.5S RNA in assembly of the bacterial signal recognition particle with its receptor. AB - The mechanism by which a signal recognition particle (SRP) and its receptor mediate protein targeting to the endoplasmic reticulum or to the bacterial plasma membrane is evolutionarily conserved. In Escherichia coli, this reaction is mediated by the Ffh/4.5S RNA ribonucleoprotein complex (Ffh/4.5S RNP; the SRP) and the FtsY protein (the SRP receptor). We have quantified the effects of 4.5S RNA on Ffh-FtsY complex formation by monitoring changes in tryptophan fluorescence. Surprisingly, 4.5S RNA facilitates both assembly and disassembly of the Ffh-FtsY complex to a similar extent. These results provide an example of an RNA molecule facilitating protein-protein interactions in a catalytic fashion. PMID- 10834843 TI - Uninterrupted MCM2-7 function required for DNA replication fork progression. AB - Little is known about the DNA helicases required for the elongation phase of eukaryotic chromosome replication. Minichromosome maintenance (MCM) protein complexes have DNA helicase activity but have only been functionally implicated in initiating DNA replication. Using an improved method for constructing conditional degron mutants, we show that depletion of MCMs after initiation irreversibly blocks the progression of replication forks in Saccharomyces cerevisiae. Like the Escherichia coli dnaB and SV40 T antigen helicases, therefore, the MCM complex is loaded at origins before initiation and is essential for elongation. Restricting MCM loading to the G(1) phase ensures that initiation and elongation occur just once per cell cycle. PMID- 10834844 TI - Essential role for cholesterol in entry of mycobacteria into macrophages. AB - Mycobacteria are intracellular pathogens that can invade and survive within host macrophages, thereby creating a major health problem worldwide. The molecular mechanisms involved in mycobacterial entry are still poorly characterized. Here we report that cholesterol is essential for uptake of mycobacteria by macrophages. Cholesterol accumulated at the site of mycobacterial entry, and depleting plasma membrane cholesterol specifically inhibited mycobacterial uptake. Cholesterol also mediated the phagosomal association of TACO, a coat protein that prevents degradation of mycobacteria in lysosomes. Thus, by entering host cells at cholesterol-rich domains of the plasma membrane, mycobacteria may ensure their subsequent intracellular survival in TACO-coated phagosomes. PMID- 10834845 TI - Lack of a role for iron in the Lyme disease pathogen. AB - A fundamental tenet of microbial pathogenesis is that bacterial pathogens must overcome host iron limitation to establish a successful infection. Surprisingly, the Lyme disease pathogen Borrelia burgdorferi has bypassed this host defense by eliminating the need for iron. B. burgdorferi grew normally and did not alter gene expression in the presence of iron chelators. Furthermore, typical bacterial iron-containing proteins were not detected in cell lysates, nor were the genes encoding such proteins identified in the genome sequence. The intracellular concentration of iron in B. burgdorferi was estimated to be less than 10 atoms per cell, well below a physiologically relevant concentration. PMID- 10834846 TI - Intracellular parasitism by the human granulocytic ehrlichiosis bacterium through the P-selectin ligand, PSGL-1. AB - Human granulocytic ehrlichiosis (HGE) is a febrile tick-borne illness caused by a recently discovered intracellular bacterium remarkable for its tropism for professionally phagocytic neutrophils. Monoclonal antibodies against the P selectin binding domain of the leukocyte P-selectin glycoprotein ligand, PSGL-1, prevented HGE cell binding and infection, as did enzymatic digestion of PSGL-1. Furthermore, simultaneous neoexpression in nonsusceptible cells of complementary DNAs for both PSGL-1 and its modifying alpha-(1,3) fucosyltransferase, Fuc-TVII, allowed binding and infection by HGE. Thus, the HGE bacterium specifically bound to fucosylated leukocyte PSGL-1. Selectin mimicry is likely central to the organism's unique ability to target and infect neutrophils. PMID- 10834847 TI - The prefrontal cortex: response selection or maintenance within working memory? AB - It is controversial whether the dorsolateral prefrontal cortex is involved in the maintenance of items in working memory or in the selection of responses. We used event-related functional magnetic resonance imaging to study the performance of a spatial working memory task by humans. We distinguished the maintenance of spatial items from the selection of an item from memory to guide a response. Selection, but not maintenance, was associated with activation of prefrontal area 46 of the dorsal lateral prefrontal cortex. In contrast, maintenance was associated with activation of prefrontal area 8 and the intraparietal cortex. The results support a role for the dorsal prefrontal cortex in the selection of representations. This accounts for the fact that this area is activated both when subjects select between items on working memory tasks and when they freely select between movements on tasks of willed action. PMID- 10834848 TI - Generalized potential of adult neural stem cells. AB - The differentiation potential of stem cells in tissues of the adult has been thought to be limited to cell lineages present in the organ from which they were derived, but there is evidence that some stem cells may have a broader differentiation repertoire. We show here that neural stem cells from the adult mouse brain can contribute to the formation of chimeric chick and mouse embryos and give rise to cells of all germ layers. This demonstrates that an adult neural stem cell has a very broad developmental capacity and may potentially be used to generate a variety of cell types for transplantation in different diseases. PMID- 10834849 TI - The first annual EULAR congress PMID- 10834850 TI - Education to improve the health of the nation: who should we educate? PMID- 10834851 TI - Committee on Publication Ethics (COPE) guidelines on good publication practice. PMID- 10834852 TI - Anti-TNFalpha: a new dimension in the pharmacotherapy of the spondyloarthropathies !? PMID- 10834853 TI - Do patients with osteoarthritis get the clinical research they need? PMID- 10834854 TI - Rheumatology: what should all doctors know? PMID- 10834855 TI - Epidemiology of research into interventions for the treatment of osteoarthritis of the knee joint. AB - OBJECTIVE: To assess the published research base for interventions for osteoarthritis of the knee, and to identify areas in need of further research. METHODS: Literature searches were conducted on electronic databases (Medline, Embase, ISI, and Cochrane library), bibliographies of existing review articles were hand searched, and a postal questionnaire was sent to members of the Osteoarthritis Research Society International. All relevant articles were copied and searched for treatment type, study methodology, statistical results, conclusions, funding source, researcher affiliations, and year of publication, using a predetermined data extraction form. RESULTS: There have been marked changes in the literature over the period studied (1950-98), with a recent rise in trials of physical therapy, educational interventions, and complementary treatments. However, overall, most research was either drug (59.1%) or surgically (25.6%) related. Most of the studies reported positive results (94%). Research on oral drugs was significantly more likely to provide a positive result than research on any other intervention (p<0.001 by chi(2) test). Commercially funded studies were significantly more likely to produce a positive result than non commercially funded research (p=0.0027 by chi(2) test). CONCLUSIONS: Analysis of time trends indicates that the research agenda does shadow changes in consumer demands. However, there are significant gaps in the research base that need to be considered. PMID- 10834856 TI - Proposal to establish a register for the long term surveillance of adverse events in patients with rheumatic diseases exposed to biological agents: the EULAR Surveillance Register for Biological Compounds. PMID- 10834857 TI - Rheumaderm: current issues in rheumatology and dermatology PMID- 10834858 TI - Radiographic assessment of hip osteoarthritis progression: impact of reading procedures for longitudinal studies. AB - OBJECTIVE: To compare radiographic reading procedures and evaluate their impact on sample size in hip osteoarthritis (OA) longitudinal studies. METHODS: Pelvic radiographs performed twice, three years apart, in 104 patients with hip OA were read by a single reader using the Kellgren and Lawrence system, joint space narrowing scale, and joint space width (JSW). Reading procedures were (a) films read as single radiographs, (b) films grouped by patient but read in random order, (c) films grouped by patient and chronologically ordered, all with landmarks for JSW measurements, (d) films read as single radiographs, without landmarks for JSW measurements. JSW was measured at the narrowest point with a 0.1 mm graduated magnifying glass. RESULTS: More Kellgren and Lawrence or joint space narrowing grades were modified respectively with the single (42% and 37%) than with the paired (32% and 23%) or chronologically ordered (34% and 29%) reading procedures. Variability of JSW progression was principally related to mean progression (88.3%) and landmarks (almost 10%). Standardised response means were -0.71 with the paired reading procedure with landmarks, -0.68 with the single reading procedure with landmarks, -0.65 with the single reading procedure without landmarks. With landmarks, 10% more patients would be needed using single than paired reading. Using single reading, 10% more patients would be needed without landmarks than with landmarks. CONCLUSION: Kellgren and Lawrence grading seems to be influenced by the reading procedure, as is joint space narrowing grading, for assessing hip OA. Paired reading procedure with landmarks for JSW should be recommended in longitudinal studies. PMID- 10834860 TI - Improved medical and surgical management of cervical spine disease in patients with rheumatoid arthritis over 10 years. AB - OBJECTIVES: (1) To compare clinical outcome and symptomatology of rheumatoid cervical myelopathy between patients managed conservatively and surgically. (2) To determine if surgical outcome has improved since the series published from this unit in 1987. (3) To examine the role of magnetic resonance imaging (MRI) in the diagnosis of cervical myelopathy. METHODS: Patients undergoing MRI of the cervical spine between 1991 and 1996 were identified. Case records were reviewed retrospectively. RESULTS: 111 patients with RA underwent 124 MRI scans. The median age at onset of cervical spine symptoms was 58 years (range 16-87) with median disease duration of 16 years (range 1-59). 18 (16%) required surgery immediately after MRI. 93 (84%) were managed conservatively, 9 of whom (10%) later required surgery. 2/7 deaths in the conservative group were directly related to cervical myelopathy. Patients requiring surgery were more likely to report paraesthesia, weakness, unsteadiness and to exhibit extensor plantar reflexes, gait disturbance, and reduced power. MRI findings did not correlate with clinical features. When compared with the 1974-82 cohort, fewer patients had severe myelopathy (Ranawat grade IIIB) before surgery (34% versus 7%). Early postoperative mortality improved from 9% to 0% and surgical complication rate fell from 50% to 22%. 89% of patients in the 1991-96 cohort reported subjective improvement in overall function. CONCLUSION: In this series surgical outcome has improved. The major factor in this more favourable outcome is probably that patients presenting with rheumatoid cervical myelopathy are now referred for surgery at an earlier stage of disease. Clinical findings correlate poorly with MRI findings, therefore clinical history should remain the key to determining the need for MRI. PMID- 10834859 TI - Effects of a loading dose regimen of three infusions of chimeric monoclonal antibody to tumour necrosis factor alpha (infliximab) in spondyloarthropathy: an open pilot study. AB - OBJECTIVE: To evaluate the efficacy and safety of a loading dose regimen of three intravenous infusions with infliximab in patients with active spondyloarthropathy. METHODS: A monocentre, open-label pilot study of 21 patients with different subtypes of spondyloarthropathy was conducted. Treatment resistant patients with active disease (fulfilling inclusion criteria) received three infusions of 5 mg/kg infliximab (at weeks 0, 2, and 6). Standard clinical assessments were performed at baseline, and on days 3, 7, and 14, and from then on every two weeks. In patients who fulfilled criteria for ankylosing spondylitis, axial assessment was performed at baseline and on days 14, 42, and 84. RESULTS: In all global assessments (visual analogue scale of patient global assessment, patient pain assessment, doctor global assessment), erythrocyte sedimentation rate, and C reactive protein, a highly significant decrease could be seen already at day 3 (compared with baseline), which was maintained up to day 84. In patients with peripheral disease (n=18), tender and swollen joint count significantly decreased. In patients with axial disease (n=11), functional and disease activity indices significantly improved. Moreover in eight patients with psoriatic arthritis a significant decrease of the psoriasis area and severity index was observed. The treatment was well tolerated in all patients; no significant adverse events were seen. CONCLUSION: In this open-label pilot study of a loading dose regimen of three infusions of chimeric monoclonal antibody to tumour necrosis factor alpha in patients with active spondyloarthropathy, there was a fast and significant improvement of axial and peripheral articular manifestations, without major adverse experiences. PMID- 10834861 TI - Interindividual and intra-articular variation of proinflammatory cytokines in patients with rheumatoid arthritis: potential implications for treatment. AB - OBJECTIVES: Assessment of the numbers and spatial distribution of cells producing interleukin 1alpha (IL1alpha), interleukin 1beta (IL1beta), tumour necrosis factor alpha (TNFalpha), and interleukin 6 (IL6) in the synovial membranes of patients with rheumatoid arthritis (RA). METHODS: Synovial tissue specimens from 40 patients with RA and eight patients with non-rheumatic disease were obtained by arthroscopy guided biopsy techniques or during joint surgery. A modified immunohistochemical method detecting cytokine producing rather than cytokine binding cells was applied to determine cytokine synthesis in fixed cryopreserved sections. Computerised image analysis methods provided comparative quantitative assessments. RESULTS: A wide variation between subjects was recorded for both quantities and profiles of expressed cytokines, despite similar macroscopic and histopathological features of inflammation. IL1alpha and IL1beta were the most abundant monokines identified, though produced at different sites. IL1alpha was predominantly seen in vascular endothelial cells, whereas IL1beta staining was mainly shown in macrophages and fibroblasts. Concordant results for the detection of TNFalpha at protein and mRNA levels were obtained with an unexpectedly low number of TNFalpha producing cells compared with IL1 expressing cells in many patients with RA. Specimens acquired arthroscopically from areas with maximum signs of macroscopic inflammation showed an increased number of TNFalpha producing cells in pannus tissue compared with that occurring in synovial villi of a given joint. This clustered distribution was not found for cells expressing any of the other studied cytokines. CONCLUSION: The recorded heterogeneous profile of proinflammatory cytokine synthesis in the synovial membrane among patients with RA may provide a clue for an understanding of the wide variation in responsiveness to different modes of antirheumatic treatment between patients. PMID- 10834862 TI - Expression of thrombospondin-1 and its receptor CD36 in human osteoarthritic cartilage. AB - OBJECTIVE: Thrombospondin-1 (TSP-1), a trimeric glycoprotein, is involved in cell matrix interactions of various tissues, particularly in cartilage. Biochemical analyses show expression of TSP-1 in human cartilage, but its cellular source as well as the presence of its main surface receptors CD36 and CD51 in normal and osteoarthritic cartilage remain unknown. Therefore, to localise TSP-1 and its receptors immunohistochemistry and in situ hybridisation were used. METHODS: Radioactive in situ hybridisations with an RNA probe that encodes TSP-1 combined with immunostaining were carried out to investigate the expression patterns of TSP-1, CD36, and CD51 in seven normal and 23 osteoarthritic human cartilage samples. RESULTS: In normal cartilage TSP-1 was present mainly in the middle and upper deep zone. RNA expression was predominantly seen over chondrocytes of the middle zone. CD36 was found in chondrocytes of the superficial and upper middle zone. In mild and moderate osteoarthritic cartilage an increased number of TSP-1 expressing chondrocytes were seen and an increased pericellular staining close to the surface. In severe osteoarthritic cartilage a decrease in the number of TSP-1 synthesising chondrocytes and a strong reduction in matrix staining were observed. Most of these severe osteoarthritic samples showed a strongly enhanced number of CD36 positive chondrocytes. CONCLUSION: The cellular source of TSP-1 in normal cartilage is mainly mid-zone chondrocytes, which also express CD36. In early osteoarthritic cartilage lesions an increase of TSP-1 was seen, whereas reduced TSP-1 synthesis is paralleled by a strong decrease in TSP-1 protein staining in severe osteoarthritis. Furthermore, in severe osteoarthritic cartilage the number of CD36 immunostained chondrocytes is significantly increased. PMID- 10834863 TI - Matrix metalloproteinases and tissue inhibitors of metalloproteinases in synovial fluids from patients with rheumatoid arthritis or osteoarthritis. AB - OBJECTIVE: Matrix metalloproteinases (MMPs) are expressed in joint tissues of patients with rheumatoid arthritis (RA) and osteoarthritis (OA). The objective of this study was to define the steady state levels of seven different MMPs and two tissue inhibitors of metalloproteinases (TIMPs) as well as the potential metalloproteinase activity in the synovial fluid (SF) to provide more insight into the role of MMPs in cartilage destruction in RA and OA. METHODS: Levels of MMP-1, MMP-2, MMP-3, MMP-7, MMP-8, MMP-9, MMP-13, TIMP-1, and TIMP-2 in SF aspirated from knee joints of 97 patients with RA and 103 patients with OA were measured by the corresponding one step sandwich enzyme immunoassays. Proteolytic activity of MMPs in these SFs was examined in an assay using [(3)H]carboxymethylated transferrin substrate in the presence of inhibitors of serine and cysteine proteinases after activation with p-aminophenylmercuric acetate (APMA). Destruction of RA knee joints was radiographically evaluated. RESULTS: Levels of MMP-1, MMP-2, MMP-3, MMP-8, and MMP-9 were significantly higher in RA SF than in OA SF. MMP-7 and MMP-13 were detectable in more than 45% of RA SFs and in less than 20% of OA SFs, respectively. Among the MMPs examined, MMP-3 levels were extremely high compared with those of other MMPs. Direct correlations were seen between the levels of MMP-1 and MMP-3 and between those of MMP-8 and MMP-9 in RA SF. Although the levels of MMP-1 and MMP-3 increased even in the early stage of RA, those of MMP-8 and MMP-9 were low in the early stage and increased with the progression of RA. Molar ratios of the total amounts of the MMPs to those of the TIMPs were 5.2-fold higher in patients with RA than in OA, which was significant. APMA-activated metalloproteinase activity in SF showed a similar result, and a direct correlation was seen between the molar ratios and the activity in RA SF. CONCLUSIONS: Our results show that high levels of MMP-1, MMP-2, MMP-3, MMP-8, MMP-9, and TIMP-1 are present in RA SF and suggest that once these MMPs are fully activated, they have an imbalance against TIMPs, which may contribute to the cartilage destruction in RA. PMID- 10834864 TI - Risk factors for avascular necrosis of bone in patients with systemic lupus erythematosus: is there a role for antiphospholipid antibodies? AB - BACKGROUND: Avascular necrosis of bone (AVN) is a well known complication in patients with systemic lupus erythematosus (SLE). OBJECTIVE: To investigate the role of antiphospholipid antibody status (IgM and IgG anticardiolipin antibodies and lupus anticoagulant) with adjustment for corticosteroid use as risk factors for the development of AVN. METHODS: A cohort of 265 patients receiving long term follow up in our SLE clinic from 1978 to 1998 was analysed. Patients with AVN complications were detected and then matched for age, sex, ethnicity, duration of disease, and organ disease with two other patients with SLE. A further 31 patients were chosen at random for the analysis. RESULTS: Eleven patients had AVN, giving a point prevalence of 4%. There were no significant differences demonstrable in the presence of individual antiphospholipid antibodies (aPL) or their combination between the group with AVN or the two control groups. CONCLUSION: Incorporating an adjustment for corticosteroid use we were unable to show a link between the presence of aPL and the development of AVN in patients with SLE. PMID- 10834865 TI - Aggressive treatment in early rheumatoid arthritis: a randomised controlled trial. On behalf of the Rheumatic Research Foundation Utrecht, The Netherlands. AB - OBJECTIVES: To compare three therapeutic strategies using slow acting antirheumatic drugs (SAARDs) in early rheumatoid arthritis (RA), for their disease modifying properties, toxicity, and lag time until treatment effect. METHODS: Patients with recent onset RA from six hospitals were randomly assigned to immediate initiation of one of three treatment strategies: (I) a "mild SAARD with a long lag time" (hydroxychloroquine, if necessary replaced by auranofin); (II) a "potent SAARD with a long lag time" (intramuscular gold, if necessary replaced by D-penicillamine); (III) a "potent SAARD with a short lag time" (methotrexate, if necessary replaced by sulfasalazine). Comparisons included two years of follow up. RESULTS: All SAARD strategies reduced mean disease activity. A greater percentage of patients improved clinically with strategies II and III than with strategy I: percentages of patients improved on joint score with strategies II and III (79% and 82%, respectively), which was statistically different from strategy I (66%). The same was true for remission percentages: 31% and 24% v 16%, respectively). Longitudinal analysis showed significantly less disability with strategy III, and a lower erythrocyte sedimentation rate with strategy II than with strategy I. In addition, radiological damage after one and two years, was significantly lower in strategies II and III (at two years median scores were 11 and 10 v 14 in strategy I, p<0.05). Toxicity was increased in strategy II compared with the other strategies. CONCLUSION: Strategy III, comprising methotrexate or sulfasalazine, produced the best results weighing effectiveness and toxicity. Strategy I (hydroxychloroquine or auranofin) was slightly less effective, and strategy II (intramuscular gold or D-penicillamine) was associated with increased toxicity. PMID- 10834866 TI - A prospective study of vasculitis patients collected in a five year period: evaluation of the Chapel Hill nomenclature. AB - OBJECTIVE: To test the usefulness of the Chapel Hill nomenclature, supplemented with surrogate parameters, as diagnostic criteria for primary vasculitides. METHODS: To prospectively evaluate vasculitis patients according to a standardised clinical and para-clinical programme. In accordance with the Chapel Hill publication surrogate parameters were used: proteinuria, haematuria and red blood cell casts (glomerulonephritis), angiographic or ultrasonic demonstration of aneurysms or stenoses (arteritis), radiological lung infiltrates or cavitations of more than one month's duration (granuloma in the lungs), bloody nasal discharge or crusts, chronic sinusitis, otitis and/or mastoiditis, bone and/or cartilage destruction, and acute hearing loss (granuloma in upper airways). RESULTS: The following entities were diagnosed: giant cell arteritis (n=14), Takayasu arteritis (n=1), polyarteritis nodosa (n=2), Wegener's granulomatosis (n=27), Churg-Strauss syndrome (n=2), microscopic polyangiitis (n=12), Henoch-Schonlein purpura (n=2), cutaneous leucocytoclastic angiitis (n=37), and secondary vasculitis (n=21). Giant cell arteritis and cutaneous leucocytoclastic angiitis were in all cases diagnosed by biopsy. Using the Chapel Hill nomenclature supplemented with surrogate parameters, only 8 of 27 patients were diagnosed with Wegener's granulomatosis, and 3 of 12 cases with microscopic polyangiitis. The number of patients in the remaining diagnostic entities were considered to few to evaluate. CONCLUSIONS: The Chapel Hill nomenclature, supplemented with surrogate parameters, failed to act as diagnostic criteria in Wegener's granulomatosis and microscopic polyangiitis. The following diagnostic criteria are proposed for Wegener's granulomatosis: (1) Biopsy or surrogate parameter for granulomatous inflammation in the respiratory system and (2) Biopsy verified necrotising vasculitis in small to medium sized vessels or biopsy/surrogate parameter for glomerulonephritis or positive PR3-ANCA test and (3) Lack of eosinophilia in blood and biopsy samples. The following diagnostic criteria are proposed for microscopic polyangiitis: (1) Biopsy verified necrotising vasculitis in small vessels and/or glomerulonephritis with few or no immune deposits and (2) Involvement of more than one organ system as indicated by biopsy verified vasculitis in small to medium sized vessels or surrogate parameter for glomerulonephritis and (3) Lack of biopsy and surrogate parameter for granulomatous inflammation in the respiratory system. Using these criteria all Wegener's patients and 9 of 12 patients with microscopic polyangiitis could be diagnosed. PMID- 10834867 TI - Anticardiolipin autoantibodies in serum samples and cryoglobulins of patients with chronic hepatitis C infection. AB - OBJECTIVE: Chronic hepatitis C virus (HCV) has been linked to extrahepatic autoimmune phenomena. In addition, a variety of autoantibodies are found in patients with HCV. The prevalence, nature, and clinical significance of anticardiolipin (aCL) autoantibodies in serum samples of patients with HCV were therefore investigated. PATIENTS AND METHODS: A prospective study of 48 consecutive patients with chronic HCV with no evidence of previous hepatitis B virus (HBV) infection or any other autoimmune disorder. Thirty patients with HBV and 50 healthy volunteers matched for age and sex served as control groups. Anticardiolipin antibodies in the serum samples and cryoprecipitates were measured by a sensitive enzyme linked immunosorbent assay (ELISA). The beta(2) glycoprotein I (beta(2)-GPI) dependency was determined by carrying out aCL assays in the presence or absence of fetal calf serum samples. RESULTS: High levels of IgG aCL antibodies were detected in serum samples of 21/48 (44%) patients with HCV. These autoantibodies showed no beta(2)-GPI dependency. The control groups had much lower levels of aCL antibodies (20% in the patients with HBV and none in the normal volunteers). Cryoprecipitates from four patients with HCV (three aCL positive; one aCL negative) were further isolated. In two of the three aCL positive patients, specific cardiolipin reactivity was shown in the cryoprecipitates. The group of patients with HCV and aCL antibodies in their serum showed significantly higher total IgG levels, a higher incidence of antinuclear antibodies, and viraemia (HCV RNA) than the aCL negative patients. None of the patients with HCV and aCL antibodies showed any clinical manifestations related to those autoantibodies. CONCLUSIONS: This study clearly shows a high prevalence of IgG aCL antibodies in the serum of patients with HCV and the localisation of these antibodies in some cryoprecipitates. The role of these autoantibodies on the course of HCV infection and their clinical significance has not yet been determined. PMID- 10834869 TI - How should we manage fibromyalgia? PMID- 10834868 TI - An audit of cyclosporin for systemic lupus erythematosus and related overlap syndromes: limitations of its use. AB - OBJECTIVE: To establish the usefulness of cyclosporin for systemic lupus erythematosus (SLE) in a routine clinical setting. METHODS: Patients who had received cyclosporin for SLE, mixed connective tissue disease, and other overlap syndromes were identified. Data relating to treatment with cyclosporin, including dosage, concurrent steroid use, response to treatment, side effects, and reasons for withdrawal, were extracted from medical notes. RESULTS: A total of 43 patients had been treated with cyclosporin between 1995 and 1998. Cyclosporin, average dose 4 mg/kg/d, was started in patients whose disease was active despite previous use of alternative second line agents. On every occasion when cyclosporin was used for thrombocytopenia the response was good, but when arthritis was the indication, the response was good in 14/26. The success rates for symptoms of arthralgia, myalgia, and fatigue were lower. Side effects occurred in 28/43 (65%) cases, and on 39/47 (83%) occasions cyclosporin was withdrawn owing to either side effects or failure to control disease activity, after a median duration of treatment of only four months. CONCLUSIONS: The response to cyclosporin is mixed and usually short lived. PMID- 10834870 TI - Adjuvant irradiation for breast cancer. PMID- 10834871 TI - Stalking: why do people do it? PMID- 10834872 TI - HIV risk behaviour in gay men: on the rise? PMID- 10834873 TI - New approaches to conversion hysteria. PMID- 10834874 TI - Revalidation for doctors in the United Kingdom: the end or the beginning? PMID- 10834875 TI - Clinton seeks heavy fines for breaching research ethics. PMID- 10834876 TI - Unusable medicines "dumped" on Venezuela. PMID- 10834878 TI - Coffee may lower risk of Parkinson's disease PMID- 10834877 TI - Asylum seekers in UK receive poor health care. PMID- 10834879 TI - In brief PMID- 10834880 TI - Mouth to mouth ventilation does not improve CPR PMID- 10834881 TI - Prisoners in england and wales are at risk of bloodborne viruses PMID- 10834882 TI - Junior doctors accept new pay deal. PMID- 10834883 TI - Romanian junior doctors left without salaries. PMID- 10834884 TI - Irish juniors defer their strike. PMID- 10834885 TI - BMA demands more responsible media attitude on body image. PMID- 10834886 TI - SLE is caused by cell debris PMID- 10834887 TI - Geographical mobility is predictor of demand on psychiatric services PMID- 10834888 TI - High insulin levels linked to deaths from breast cancer. PMID- 10834890 TI - Relation between troponin T concentration and mortality in patients presenting with an acute stroke: observational study. AB - OBJECTIVE: To assess whether a raised serum troponin T concentration would be an independent predictor of death in patients with an acute ischaemic stroke. DESIGN: Observational study. SETTING: Auckland Hospital, Auckland, New Zealand. SUBJECTS: All 181 patients with an acute ischaemic stroke admitted over nine months in 1997-8, from a total of 8057 patients admitted to the acute medical service. MAIN OUTCOME MEASURES: Blood samples for measuring troponin T concentration were collected 12-72 hours after admission; other variables previously associated with severity of stroke were also recorded and assessed as independent predictors of inpatient mortality. RESULTS: Troponin T concentration was raised (>0.1 microgram/l) in 17% (30) of patients admitted with an acute ischaemic stroke. Thirty one patients died in hospital (12/30 (40%) patients with a raised troponin T concentration v 19/151 (13%) patients with a normal concentration (relative risk 3.2 (95% confidence 1.7 to 5. 8; P=0.0025)). Of 17 possible predictors of death, assessed in a multivariate stepwise model, only a raised troponin T concentration (P=0.0002), age (P=0.0008), and an altered level of consciousness at presentation (P=0.0074) independently predicted an adverse outcome. CONCLUSIONS: Serum troponin T concentration at hospital admission is a powerful predictor of mortality in patients admitted with an acute ischaemic stroke. PMID- 10834889 TI - Human T cell leukaemia/lymphoma virus infection in pregnant women in the United Kingdom: population study. AB - OBJECTIVE: To assess the prevalence of human T cell leukaemia/lymphoma virus (HTLV) infection in pregnant women in the United Kingdom. DESIGN: Population study. SUBJECTS: Guthrie card samples from babies born in 1997-8. Samples were linked to data on mother's age and ethnic status and parents' country of birth and then anonymised. SETTING: North Thames Regional Health Authority. MAIN OUTCOME MEASURES: Presence of antibodies against HTLV in eluates tested by gelatin particle agglutination assay and results confirmed by immunoblot. RESULTS: Of 126 010 samples tested, 67 had confirmed antibodies to HTLV (59 HTLV I, 2 HTLV-II, 6 untyped) and six had indeterminate results. Seroprevalence was 17.0 per 1000 (95% confidence interval 9.2 to 28.3) in infants whose mothers were born in the Caribbean, 3.2/1000 (1.5 to 5.9) with mothers born in west and central Africa, and 6.8/1000 (3.1 to 12.9) in infants of black Caribbean mothers born in non-endemic regions. In infants with no known risk (both parents born in non-endemic regions and mother not black Caribbean) seroprevalence was 0.06-0.12 per 1000. Mother's country of birth, father's country of birth, and mother's ethnic status were all independently associated with neonatal seroprevalence. An estimated 223 (95% confidence interval 110 to 350) of the 720 000 pregnant women each year in the United Kingdom are infected with HTLV. CONCLUSIONS: The prevalence of HTLV and HIV infections in pregnant women in the United Kingdom are comparable. The cost effectiveness of antenatal HTLV screening should be evaluated, and screening of blood donations should be considered. PMID- 10834892 TI - Increase in high risk sexual behaviour among homosexual men, London 1996-8: cross sectional, questionnaire study. PMID- 10834891 TI - Single blind, randomised trial of efficacy and acceptability of oral picolax versus self administered phosphate enema in bowel preparation for flexible sigmoidoscopy screening. AB - OBJECTIVE: To compare the acceptability and efficacy of two methods of self administered bowel preparation for flexible sigmoidoscopy screening: a single phosphate enema and a single sachet of Picolax. DESIGN: Single blind, randomised trial. SETTING: Endoscopy units of two general hospitals. PARTICIPANTS: 1442 men and women aged 55-64 years who had agreed to be screened by flexible sigmoidoscopy. MAIN OUTCOME MESURESs: Attendance rates, compliance with allocated preparations, adverse effects, quality of bowel preparation, procedure time, and yield of neoplasia. RESULTS: Compliance with the enema was higher than with the Picolax (608 (84%) v 566 (79%); difference 6%, 95% confidence interval 2% to 10%). Almost half of those who refused Picolax used an enema at home. Wind, incontinence, and sleep disturbance were more frequent in the Picolax group than the enema group; bottom soreness was more frequent in the enema group. Around 30% (187) found the diet restriction required by Picolax difficult; 78% (471) found the enema easy to administer. The quality of preparation was better with the enema; the proportion of procedures complete to the descending colon was greater and the mean duration of the procedure was shorter. There was no significant difference in polyp detection rates. CONCLUSION: A single phosphate enema self administered around one hour before leaving home is a more acceptable and effective method of preparing the distal bowel for flexible sigmoidoscopy than Picolax. PMID- 10834893 TI - Retrospective case note review of acute and inpatient stroke outcomes. PMID- 10834895 TI - A wise physician PMID- 10834896 TI - Me, the heartsink patient PMID- 10834894 TI - Preventing dog bites in children: randomised controlled trial of an educational intervention. PMID- 10834897 TI - Asthma after childhood pneumonia: six year follow up study. AB - OBJECTIVE: To establish the long term cumulative prevalence of asthma in children admitted to hospital with pneumonia and to examine the hypothesis that some children admitted to hospital with pneumonia may be presenting with undiagnosed asthma. DESIGN: Prospective study of a cohort of children previously admitted to hospital with pneumonia, followed up by postal questionnaires to their general practitioners and the children or their parents. SETTING: General practices in southwest England. PARTICIPANTS: 78 children admitted to the Royal Devon and Exeter Hospital between 1989 and 1991 with a diagnosis of pneumonia confirmed on independent review of x ray films. MAIN OUTCOME MEASURES: Any diagnosis of asthma, use of any treatment for asthma, and asthma symptom scores. RESULTS: On the basis of a 100% response rate from general practitioners and 86% from patients or parents, the cumulative prevalence of asthma was 45%. A diagnosis of asthma was associated with a family history of asthma (odds ratio 11.23; 95% confidence interval 2.57 to 56.36; P=0.0002). Mean symptom scores were higher for all children with asthma (mean score 2.4; chi(2)=14.88; P=0. 0001) and for children with asthma not being treated (mean 1.4; chi(2)=6.2; P=0.01) than for those without asthma (mean 0.2). CONCLUSIONS: A considerable proportion of children presenting to a district general hospital with pneumonia either already have unrecognised asthma or subsequently develop asthma. The high cumulative prevalence of asthma suggests that careful follow up of such children is worth while. Asthma is undertreated in these children; a structured symptom questionnaire may help to identify and reduce morbidity due to undertreatment. PMID- 10834898 TI - We are interdependent PMID- 10834900 TI - Management of lateral neck masses in adults. PMID- 10834899 TI - Systematic review of studies of patient satisfaction with telemedicine. AB - OBJECTIVE: To review research into patient satisfaction with teleconsultation, specifically clinical consultations between healthcare providers and patients involving real time interactive video. DESIGN: Systematic review of telemedicine satisfaction studies. Electronic databases searched include Medline, Embase, Science Citation Index, Social Sciences Citation Index, Arts and Humanities Citation Index, and the TIE (Telemedicine Information Exchange) database. SUBJECTS: Studies conducted worldwide and published between 1966 and 1998. MAIN OUTCOME MEASURES: Quality of evidence about patient satisfaction. RESULTS: 32 studies were identified. Study methods used were simple survey instruments (26 studies), exact methods not specified (5), and qualitative methods (1). Study designs were randomised controlled trial (1 trial); random patient selection (2); case-control (1); and selection criteria not specified or participants represented consecutive referrals, convenience samples, or volunteers (28). Sample sizes were 100 (7), and not specified (1). All studies reported good levels of patient satisfaction. Qualitative analysis revealed methodological problems with all the published work. Even so, important issues were highlighted that merit further investigation. There is a paucity of data examining patients' perceptions or the effects of this mode of healthcare delivery on the interaction between providers and clients. CONCLUSIONS: Methodological deficiencies (low sample sizes, context, and study designs) of the published research limit the generalisability of the findings. The studies suggest that teleconsultation is acceptable to patients in a variety of circumstances, but issues relating to patient satisfaction require further exploration from the perspective of both clients and providers. PMID- 10834902 TI - What is humanity? PMID- 10834901 TI - Persistent diarrhoea and occult vipomas in children. PMID- 10834904 TI - An old friend PMID- 10834903 TI - Swollen lower limb-2: lymphoedema. PMID- 10834905 TI - Using conjoint analysis to elicit preferences for health care. PMID- 10834906 TI - Challenges to the health services: the professions. PMID- 10834908 TI - Obituaries PMID- 10834907 TI - Triage PMID- 10834909 TI - Breast cancer: sharing the decision PMID- 10834910 TI - Resuscitation rules PMID- 10834911 TI - Peer review in health sciences PMID- 10834912 TI - Defining features PMID- 10834913 TI - The sum of my parts PMID- 10834914 TI - Gay sex PMID- 10834915 TI - One person's innovation is another's experiment. PMID- 10834916 TI - Netlines PMID- 10834917 TI - Railway signals passed at danger: psychology matters PMID- 10834918 TI - Choosing socks PMID- 10834921 TI - Troponin T predicts mortality in patients with ischaemic stroke PMID- 10834919 TI - Time to reinvent yourself? PMID- 10834920 TI - HTLV is as prevalent as HIV among UK infants PMID- 10834922 TI - Unsafe sexual practices are increasing among gay men in london PMID- 10834923 TI - Patients prefer enemas to laxatives for bowel preparation PMID- 10834925 TI - Studies of patient satisfaction with telemedicine are inconclusive PMID- 10834924 TI - People who have strokes while in hospital have a poor prognosis PMID- 10834926 TI - Peptide nucleic acid (PNA): its medical and biotechnical applications and promise for the future. AB - Synthetic molecules that can bind with high sequence specificity to a chosen target in a gene sequence are of major interest in medicinal and biotechnological contexts. They show promise for the development of gene therapeutic agents, diagnostic devices for genetic analysis, and as molecular tools for nucleic acid manipulations. Peptide nucleic acid (PNA) is a nucleic acid analog in which the sugar phosphate backbone of natural nucleic acid has been replaced by a synthetic peptide backbone usually formed from N-(2-amino-ethyl)-glycine units, resulting in an achiral and uncharged mimic. It is chemically stable and resistant to hydrolytic (enzymatic) cleavage and thus not expected to be degraded inside a living cell. PNA is capable of sequence-specific recognition of DNA and RNA obeying the Watson-Crick hydrogen bonding scheme, and the hybrid complexes exhibit extraordinary thermal stability and unique ionic strength effects. It may also recognize duplex homopurine sequences of DNA to which it binds by strand invasion, forming a stable PNA-DNA-PNA triplex with a looped-out DNA strand. Since its discovery, PNA has attracted major attention at the interface of chemistry and biology because of its interesting chemical, physical, and biological properties and its potential to act as an active component for diagnostic as well as pharmaceutical applications. In vitro studies indicate that PNA could inhibit both transcription and translation of genes to which it has been targeted, which holds promise for its use for antigene and antisense therapy. However, as with other high molecular mass drugs, the delivery of PNA, involving passage through the cell membrane, appears to be a general problem. PMID- 10834927 TI - Beneficial effects of peroxynitrite decomposition catalyst in a rat model of splanchnic artery occlusion and reperfusion. AB - The aim of the present study was to investigate the protective effect of the peroxynitrite decomposition catalyst 5,10,15, 20-tetrakis(2,4,6-trimethyl-3,5 disulfonatophenyl)-porphyrinato iron (III) (FeTMPS) in a model of splanchnic artery occlusion shock (SAO). SAO shock was induced in rats by clamping both the superior mesenteric artery and the celiac trunk for 45 min, followed by release of the clamp (reperfusion). At 60 min after reperfusion, animals were killed for histological examination and biochemical studies. There was a marked increase in the oxidation of dihydrorhodamine 123 to rhodamine (a marker of peroxynitrite induced oxidative processes) in the plasma of the SAO-shocked rats after reperfusion, but not during ischemia alone. Immunohistochemical examination demonstrated a marked increase in the immunoreactivity to nitrotyrosine, an index of nitrogen species such as peroxynitrite, in the necrotic ileum in shocked rats. SAO-shocked rats developed a significant increase of tissue myeloperoxidase and malonaldehyde activity, and marked histological injury to the distal ileum. SAO shock was also associated with a significant mortality (0% survival at 2 h after reperfusion). Reperfused ileum tissue sections from SAO-shocked rats showed positive staining for P-selectin localized mainly in the vascular endothelial cells. Ileum tissue sections obtained from SAO-shocked rats and stained with antibody to ICAM-1 showed a diffuse staining. Administration of FeTMPS significantly reduced ischemia/reperfusion injury in the bowel, and reduced lipid and the production of peroxynitrite during reperfusion. Treatment with PN catalyst also markedly reduced the intensity and degree of P-selectin and ICAM-1 staining in tissue sections from SAO-shocked rats and improved survival. Our results clearly demonstrate that peroxynitrite decomposition catalysts exert a protective effect in SAO and that this effect may be due to inhibition of the expression of adhesion molecules and the tissue damage associated with peroxynitrite-related pathways. PMID- 10834928 TI - Nucleotide excision repair gene (ERCC1) deficiency causes G(2) arrest in hepatocytes and a reduction in liver binucleation: the role of p53 and p21. AB - A wide range of DNA lesions, both UV and chemically induced, are dealt with by the nucleotide excision repair (NER) pathway. Defects in NER result in human syndromes such as xeroderma pigmentosum (XP), where there is a 1000-fold increased incidence of skin cancer. The ERCC1 protein is essential for NER, but ERCC1 knockout mice are not a model for XP. In the absence of exogenous DNA damaging agents, these mice are runted and die before weaning, with dramatically accelerated liver polyploidy and elevated levels of p53. Here we present a morphological, immunological, and molecular study to understand the mechanism for the unusual liver pathology in ERCC1-deficient mice. We show that the enlarged ERCC1-deficient hepatocytes are arrested in G(2) and that DNA replication and the normal process of binucleation are both reduced. This is associated with a p53 independent increase in expression of the cyclin-dependent kinase inhibitor p21. The most dramatic feature of the ERCC1-deficient liver phenotype, the accelerated polyploidy, is not rescued by p53 deficiency, but we show that p53 is responsible for the reduced DNA replication and binucleation. We consider that the liver phenotype is a response to unrepaired endogenous DNA damage, which may reflect an additional non-NER-related function for the ERCC1 protein. PMID- 10834929 TI - Proteolytic cleavage of phospholipase C-gamma1 during apoptosis in Molt-4 cells. AB - Apoptosis is a cell suicide mechanism that requires the activation of cellular death proteases for its induction. We examined whether the progress of apoptosis involves cleavage of phospholipase C-gamma1 (PLC-gamma1), which plays a pivotal role in mitogenic signaling pathway. Pretreatment of T leukemic Molt-4 cells with PLC inhibitors such as U-73122 or ET-18-OCH(3) potentiated etoposide-induced apoptosis in these cells. PLC-gamma1 was fragmented when Molt-4 cells were treated with several apoptotic stimuli such as etoposide, ceramides, and tumor necrosis factor alpha. Cleavage of PLC-gamma1 was blocked by overexpression of Bcl-2 and by specific inhibitors of caspases such as Z-DEVD-CH(2)F and YVAD-cmk. Purified caspase-3 and caspase-7, group II caspases, cleaved PLC-gamma1 in vitro and generated a cleavage product of the same size as that observed in vivo, suggesting that PLC-gamma1 is cleaved by group II caspases in vivo. From point mutagenesis studies, Ala-Glu-Pro-Asp(770) was identified to be a cleavage site within PLC-gamma1. Epidermal growth factor receptor (EGFR) -induced tyrosine phosphorylation of PLC-gamma1 resulted in resistance to cleavage by caspase-3 in vitro. Furthermore, cleaved PLC-gamma1 could not be tyrosine-phosphorylated by EGFR in vitro. In addition, tyrosine-phosphorylated PLC-gamma1 was not significantly cleaved during etoposide-induced apoptosis in Molt-4 cells. This suggests that the growth factor-induced tyrosine phosphorylation may suppress apoptosis-induced fragmentation of PLC-gamma1. We provide evidence for the biochemical relationship between PLC-gamma1-mediated signal pathway and apoptotic signal pathway, indicating that the defect of PLC-gamma1-mediated signaling pathway can facilitate an apoptotic progression. PMID- 10834930 TI - Catecholamines up-regulate lipopolysaccharide-induced IL-6 production in human microvascular endothelial cells. AB - The catecholamine-mediated modulation of the cytokine network has primarily been demonstrated for leukocytes. Whereas catecholamines decrease the LPS-induced production of IL-6 by leukocytes, serum levels of IL-6 are dramatically increased by the catecholamine epinephrine in animal endotoxemia models. We now demonstrate that epinephrine as well as norepinephrine can induce IL-6 in an endothelial cell line (HMEC-1). Furthermore, these catecholamines could even potentiate the LPS induced IL-6 protein production. The synergistic effect of catecholamines and LPS could be reproduced in primary human skin microvascular endothelial cells. The catecholamine-induced IL-6 stimulation is based on increased IL-6 mRNA levels. RNA stability assays revealed that this regulation is not a result of enhanced RNA stability and therefore is most likely due to an increased transcription. Treatment with cycloheximide indicated that new protein synthesis is not necessary for this transcriptional up-regulation of IL-6 mRNA. Preincubation with alpha and beta receptor antagonists showed that the effect is mediated by beta(1) and beta(2)-adrenergic receptors. Thus, endothelial cells might be a possible source of increased IL-6 production observed in situations such as stress or septic shock, in which catecholamines are elevated due to endogenous production or exogenous application. PMID- 10834931 TI - In utero diethylstilbestrol (DES) exposure alters Hox gene expression in the developing mullerian system. AB - Diethylstilbestrol (DES) was widely used to treat pregnant women through 1971. The reproductive tracts of their female offspring exposed to DES in utero are characterized by anatomic abnormalities. Here we show that DES administered to mice in utero produces changes in the expression pattern of several Hox genes that are involved in patterning of the reproductive tract. DES produces posterior shifts in Hox gene expression and homeotic anterior transformations of the reproductive tract. In human uterine or cervical cell cultures, DES induces HOXA9 or HOXA10 gene expression, respectively, to levels approximately twofold that induced by estradiol. The DES-induced expression is not inhibited by cyclohexamide. Estrogens are novel morphogens that directly regulate the expression pattern of posterior Hox genes in a manner analogous to retinoic acid regulation of anterior Hox genes. Alterations in HOX gene expression are a molecular mechanism by which DES affects reproductive tract development. Changes in Hox gene expression are a potential marker for the effects of in utero drug use that may become apparent only at late stages of development. PMID- 10834932 TI - The transactivation-competent carboxyl-terminal domain of AF-9 is expressed within a sexually dimorphic transcript in rat pituitary. AB - We have investigated the biological role of the cellular counterpart of the leukemogenic AF-9 gene by cloning the rat AF-9 (rAF-9) cDNA and defining the regulation of an anterior pituitary-specific rAF-9 transcript that is expressed in a sexually dimorphic manner. Expression of this transcript is down-regulated after puberty in females and can be subsequently up-regulated in adults by ovariectomy. Hormone replacement studies have provided direct evidence that rAF-9 mRNA expression is suppressed by estrogen. Mapping the 1.9 kb anterior pituitary transcript has shown that it corresponds in size to the rAF-9 cDNA clone, which contains an open reading frame (ORF) that is truncated compared with the human AF 9 ORF, but encodes a previously defined transcriptional activation domain. Thus, the cellular AF-9 gene is alternatively expressed in a manner that reflects the presence of translocated, functionally active (oncogenic) AF-9 sequences in leukemias. Using a novel antisera raised against a rAF-9 peptide, we have also demonstrated tissue- and sex-specific expression of a nuclear 41 kDa anterior pituitary protein and have localized this protein to a major population of growth hormone synthesizing cells. By localizing the expression and defining the physiological regulation of rAF-9, our studies have provided novel insights into the AF-9 gene that will facilitate an understanding of both oncogenic and endocrine roles. PMID- 10834933 TI - Reduced glucose uptake precedes insulin signaling defects in adipocytes from heterozygous GLUT4 knockout mice. AB - Decreased GLUT4 expression, impaired insulin receptor (IR), IRS-1, and pp60/IRS-3 tyrosine phosphorylation are characteristics of adipocytes from insulin-resistant animal models and obese NIDDM humans. However, the sequence of events leading to the development of insulin signaling defects and the significance of decreased GLUT4 expression in causing adipocyte insulin resistance are unknown. The present study used male heterozygous GLUT4 knockout mice (GLUT4(+/-)) as a novel model of diabetes to study the development of insulin signaling defects in adipocytes with the progression of whole body insulin resistance and diabetes. Male GLUT4(+/-) mice with normal fed glycemia and insulinemia (N/N), normal fed glycemia and hyperinsulinemia (N/H), and fed hyperglycemia with hyperinsulinemia (H/H) exist at all ages. The expression of GLUT4 protein and the maximal insulin-stimulated glucose transport was 50% decreased in adipocytes from all three groups. Insulin signaling was normal in N/N adipose cells. From 35 to 70% reductions in insulin stimulated tyrosine phosphorylation of IR, IRS-1, and pp60/IRS-3 were noted with no changes in the cellular content of IR, IRS-1, and p85 in N/H adipocytes. Insulin-stimulated protein tyrosine phosphorylation was further decreased to 12 23% in H/H adipose cells accompanied by 42% decreased IR and 80% increased p85 expression. Insulin-stimulated, IRS-1-associated PI3 kinase activity was decreased by 20% in N/H and 68% reduced in H/H GLUT4(+/-) adipocytes. However, total insulin-stimulated PI3 kinase activity was normal in H/H GLUT4(+/-) adipocytes. Taken together, these results strongly suggest that hyperinsulinemia triggers a reduction of IR tyrosine kinase activity that is further exacerbated by the appearance of hyperglycemia. However, the insulin signaling cascade has sufficient plasticity to accommodate significant changes in specific components without further reducing glucose uptake. Furthermore, the data indicate that the cellular content of GLUT4 is the rate-limiting factor in mediating maximal insulin-stimulated glucose uptake in GLUT4(+/-) adipocytes. PMID- 10834934 TI - Co-localization of TFF3 peptide and oxytocin in the human hypothalamus. AB - TFF-peptides (formerly P domain peptides, trefoil factors) are typical secretory products of many mucous epithelial cells. TFF3 is also synthesized in the hypothalamus and has anxiolytic or anxiogenic activities when injected into the rat amygdala. Here we show by immunohistochemistry that TFF3 is localized to a distinct population of neurons of the human hypothalamic paraventricular and supraoptic nuclei. Generally, TFF3-positive cells are co-localized in oxytocin producing cells and not in vasopressin-producing cells. Relatively large amounts of TFF3-but not TFF1 and TFF2-are present in the posterior lobe of the human pituitary, where it is probably released into the bloodstream. Furthermore, TFF3 was also detectable in human postmortem cerebrospinal fluid. PMID- 10834935 TI - Regulation of adiposity by dietary calcium. AB - Recent data from this laboratory demonstrate that increasing adipocyte intracellular Ca(2+) results in a coordinated stimulation of lipogenesis and inhibition of lipolysis. We have also noted that increasing dietary calcium of obese patients for 1 year resulted in a 4.9 kg loss of body fat (P<0.01). Accordingly, we tested the possibility that calcitrophic hormones may act on adipocytes to increase Ca(2+) and lipid metabolism by measuring the effects of 1, 25-(OH)(2)-D in primary cultures of human adipocytes, and found significant, sustained increases in intracellular Ca(2+) and a corresponding marked inhibition of lipolysis (EC(50) approximately 50 pM; P<0.001), suggesting that dietary calcium could reduce adipocyte mass by suppressing 1,25-(OH)(2)-D. To test this hypothesis, we placed transgenic mice expressing the agouti gene specifically in adipocytes on a low (0.4%) Ca/high fat/high sucrose diet either unsupplemented or with 25 or 50% of the protein replaced by non-fat dry milk or supplemented to 1.2% Ca with CaCO(3) for 6 wk. Weight gain and fat pad mass were reduced by 26 39% by the three high calcium diets (P<0.001). The high calcium diets exerted a corresponding 51% inhibition of adipocyte fatty acid synthase expression and activity (P<0.002) and stimulation of lipolysis by 3. 4- to 5.2-fold (P<0.015). This concept of calcium modulation of adiposity was further evaluated epidemiologically in the NHANES III data set. After controlling for energy intake, relative risk of being in the highest quartile of body fat was set to 1.00 for the lowest quartile of Ca intake and was reduced to 0.75, 0.40, and 0.16 for the second, third, and fourth quartiles, respectively, of calcium intake for women (n=380;P<0.0009); a similar inverse relationship was also noted in men (n=7114; P<0.0006). Thus, increasing dietary calcium suppresses adipocyte intracellular Ca(2+) and thereby modulates energy metabolism and attenuates obesity risk. PMID- 10834936 TI - Entamoeba histolytica disturbs the tight junction complex in human enteric T84 cell layers. AB - Entamoeba (E.) histolytica trophozoites initiate amebiasis through invasion into the enteric mucosa. It was our aim to understand the molecular interactions between amebic trophozoites and enterocytes during the early steps of invasion. Trophozoites of E. histolytica strain HM1:IMSS were seeded on the apical side of enteric T84 cell layers, which were established on filters in two-compartment culture chambers. Cocultures were analyzed for paracellular permeability by measurement of transepithelial electrical resistance (TER) and for the tight junction proteins ZO-1, ZO-2, occludin, and cingulin by immunocytochemistry and immunoprecipitation. On direct contact with the apical side of the enteric cells, trophozoites caused an increase in paracellular permeability as evidenced by a decrease of TER associated with an increase in [(3)H]mannitol flux. Immunoprecipitation of cocultures revealed dephosphorylation of ZO-2, loss of ZO 1 from ZO-2, and degradation of ZO-1 but less so of ZO-2 and none of occludin or E-cadherin. In conclusion, trophozoite-associated increase in paracellular permeability of enteric cell layers is ascribed to disturbance of the molecular organization of tight junction proteins. PMID- 10834937 TI - TGF-beta autocrine loop regulates cell growth and myogenic differentiation in human rhabdomyosarcoma cells. AB - Transforming growth factor beta (TGF) is a well-known inhibitor of myogenic differentiation as well as an autocrine product of rhabdomyosarcoma cells. We studied the role of the TGF-beta autocrine loop in regulating growth and myogenic differentiation in the human rhabdomyosarcoma cell line, RD. We previously reported that the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) induces growth arrest and myogenic differentiation in these cells, which constitutively express muscle regulatory factors. We show that TPA inhibits the activation of secreted latent TGF-beta, thus decreasing the concentration of active TGF-beta to which the cells are exposed. This event is mediated by the TPA induced alteration of the uPA/PAI serine-protease system. Complete removal of TGF beta, mediated by the ectopic expression of a soluble type II TGF-beta receptor dominant negative cDNA, induces growth arrest, but does not trigger differentiation. In contrast, a reduction in the TGF-beta concentration, to a range of 0.14-0.20 x 10(-2) ng/ml (which is similar to that measured in TPA treated cells), mimics TPA-induced differentiation. Taken together, these data demonstrate that cell growth and suppression of differentiation in rhabdomyosarcoma cells require overproduction of active TGF-beta; furthermore, they show that a 'critical' concentration of TGF-beta is necessary for myogenic differentiation to occur, whereas myogenesis is abolished below and above this concentration. By impairing the TGF-beta autocrine loop, TPA stabilizes the factor concentration within the range compatible for differentiation to occur. In contrast, in human primary muscle cells a much higher concentration of exogenous TGF-beta is required for the differentiation inhibitory effect and TPA inhibits differentiation in these cells probably through a TGF-beta independent mechanism. These data thus clarify the mechanism underlying the multiple roles of TGF-beta in the regulation of both the transformed and differentiated phenotype. PMID- 10834938 TI - Activation of NF-kappaB p50/p65 is regulated in the developing mammary gland and inhibits STAT5-mediated beta-casein gene expression. AB - The NF-kappaB family of transcription factors regulates diverse cellular functions such as immune response and cell growth and development, and has been reported to be constitutively active in a variety of mammary carcinoma cell lines. However, its role in normal mammary gland development has not been addressed. In our study, we detected developmentally regulated NF-kappaB activity in the mammary gland of mice. During pregnancy, DNA binding activity of NF-kappaB p50/p65 increased until day 16 postcoitum and decreased with the onset of lactation, most likely due to reduced p50 and p65 protein levels in the nucleus. Cotransfection experiments performed with 293 cells revealed an inhibition of the prolactin receptor/JAK2/STAT5 pathway by NF-kappaB. In HC11 cells, NF-kappaB p50/p65 activity was inversely correlated with prolactin-induced STAT5 tyrosine phosphorylation, expression of endogenous beta-casein gene, and of a transfected beta-casein gene promoter reporter construct. This indicates a negative cross talk between NF-kappaB and the prolactin receptor/JAK2/STAT5 activation pathway, which occurs at the level of STAT5 tyrosine phosphorylation. Our results provide evidence for a role of NF-kappaB in normal mammary gland development, and indicate its function as a negative regulator of beta-casein gene expression during pregnancy by interfering with STAT5 tyrosine phosphorylation. PMID- 10834939 TI - Functional characterization of the NCC27 nuclear protein in stable transfected CHO-K1 cells. AB - NCC27 belongs to a family of small, highly conserved, organellar ion channel proteins. It is constitutively expressed by native CHO-K1 and dominantly localized to the nucleus and nuclear membrane. When CHO-K1 cells are transfected with NCC27-expressing constructs, synthesized proteins spill over into the cytoplasm and ion channel activity can then be detected on the plasma as well as nuclear membrane. This provided a unique opportunity to directly compare electrophysiological characteristics of the one cloned channel, both on the nuclear and cytoplasmic membranes. At the same time, as NCC27 is unusually small for an ion channel protein, we wished to directly determine whether it is a membrane-resident channel in its own right. In CHO-K1 cells transfected with epitope-tagged NCC27 constructs, we have demonstrated that the NCC27 conductance is chloride dependent and that the electrophysiological characteristics of the channels are essentially identical whether expressed on plasma or nuclear membranes. In addition, we show that a monoclonal antibody directed at an epitope tag added to NCC27 rapidly inhibits the ability of the expressed protein to conduct chloride, but only when the antibody has access to the tag epitope. By selectively tagging either the amino or carboxyl terminus of NCC27 and varying the side of the membrane from which we record channel activity, we have demonstrated conclusively that NCC27 is a transmembrane protein that directly forms part of the ion channel and, further, that the amino terminus projects outward and the carboxyl terminus inward. We conclude that despite its relatively small size, NCC27 must form an integral part of an ion channel complex. PMID- 10834940 TI - Novel functional PI 3-kinase antagonists inhibit cell growth and tumorigenicity in human cancer cell lines. AB - New efforts in cancer therapy are being focused at various levels of signaling pathways. With phosphoinositide 3-kinase (PI3-K) potentially being necessary for a range of cancer-related functions, we have investigated the influence of selected inositol tris- to hexakisphosphates on cell growth and tumorigenicity. We show that micromolar concentrations of inositol 1,3,4,5,6-pentakisphosphate and inositol 1,4,5,6-tetrakisphosphate [Ins(1,4,5,6)P(4)] inhibit IGF-1-induced [(3)H]-thymidine incorporation in human breast cancer (MCF-7) cells and the ability to grow in liquid medium and form colonies in agarose semisolid medium by small cell lung cancer (SCLC) cells, a human cancer cell line containing a constitutively active PI3-K. In an ovarian cancer cell line that also contains a constitutively active PI3-K (SKOV-3 cells), Ins(1,4,5,6)P(4) again inhibited liquid medium growth. Furthermore, when applied extracellularly, inositol 1,3,4,5 tetrakisphosphate was shown indeed to enter SCLC cells. These effects appeared specifically related to PH domains known to bind to phosphatidylinositol 3,4 bisphosphate [PtdIns(3,4)P(2)] and phosphatidylinositol 3,4,5-trisphosphate [PtdIns(3,4,5)P(3)], indicating involvement of the PI3-K downstream target protein kinase B (PKB/Akt). This was further supported by inhibition of PKB/Akt PH domain membrane targeting in COS-7 cells by Ins(1,4,5,6)P(4). Thus, we propose that specific inositol polyphosphates inhibit PI3-K by competing with PtdIns(3,4, 5)P(3)-binding PH domains and that this occurs mainly at the level of the downstream PI3-K target, PKB/Akt. PMID- 10834941 TI - Expression of prostaglandin endoperoxide H synthase-2 induced by nitric oxide in conditionally immortalized murine colonic epithelial cells. AB - Increased expression of prostaglandin endoperoxide H synthase-2 (PGHS-2) has been implicated in pathological conditions such as inflammatory bowel diseases and colon cancer. Recently, it has been demonstrated that inducible nitric oxide synthase (NOS II) expression and nitric oxide (NO) production are up-regulated in these diseases as well. However, the apparent link between PGHS-2 and NOS II has not been thoroughly investigated in nontransformed and nontumorigenic colonic epithelial cells. In the present study, we examined the concomitant expression of PGHS-2 and NOS II as well as the production of prostaglandin E2 (PGE2) and NO in conditionally immortalized mouse colonic epithelial cells, namely YAMC (Apc(+/+)). We found that the induction of PGHS-2 and generation of PGE2 in these cells by IFN-gamma and lipopolysaccharide (LPS) were greatly reduced by two selective NOS II inhibitors, L-NIL and SMT. To ascertain the effect of NO on PGHS 2 overexpression, we tested NO-releasing compounds, NOR-1 and SNAP, and found that they caused PGHS-2 expression and PGE2 production. This effect was abolished by hemoglobin, a NO scavenger. Using electrophoretic mobility shift assays, we found that both NOR-1 and SNAP caused beta-catenin/LEF-1 DNA complex formation. Super-shift by anti-beta-catenin antibody confirmed the presence of beta-catenin in the complex. Cell fractionation studies indicated that NO donors caused an increase in free soluble cytoplasmic beta-catenin. This is further corroborated by the immunocytochemistry data showing the redistribution of beta-catenin from the predominantly membrane localization into the cytoplasm and nucleus after treatment with NO donors. To further explore the possible connection between PGHS 2 expression and beta-catenin/LEF-1 DNA complex formation, we studied IMCE (Apc(Min/+)) cells, a sister cell line of YAMC with similar genetic background but differing in Apc genotype and, consequently, their beta-catenin levels. We found that IMCE cells, in comparison with YAMC cells, had markedly higher beta catenin/LEF-1 DNA complex formation under both resting conditions as well as after induction with NO. In parallel fashion, IMCE cells expressed significantly higher levels of PGHS-2 mRNA and protein, and generated more PGE2. Overall, this study suggests that NO may be involved in PGHS-2 overexpression in conditionally immortalized mouse colonic epithelial cells. Although the molecular mechanism of the link is still under investigation, this effect of NO appears directly or indirectly to be a result of the increase in free soluble beta-catenin and the formation of nuclear beta-catenin/LEF-1 DNA complex. PMID- 10834942 TI - Mechanisms of antiapoptotic effects of estrogens in nigral dopaminergic neurons. AB - Parkinson's disease is characterized by the mesencephalic dopaminergic neuronal loss, possibly by apoptosis, and the prevalence is higher in males than in females. The estrogen receptor (ER) subtype in the mesencephalon is exclusively ER beta, a recently cloned novel subtype. Bound with estradiol, it enhances gene transcription through the estrogen response element (ERE) or inhibits it through the activator protein-1 (AP-1) site. We demonstrated that 17beta-estradiol provided protection against nigral neuronal apoptosis caused by exposure to either bleomycin sulfate (BLM) or buthionine sulfoximine (BSO). BLM and BSO induced nigral apoptosis was blocked by inhibitors for caspase-3 or c-Jun/AP-1. The antiapoptotic effect by estradiol was blocked by ICI 182,780, an antagonist for ER, but not by a synthesized peptide that inhibits binding of the ER to the ERE. Estradiol had no effects on caspase-3 activation and c-Jun NH(2)-terminal kinase (JNK), which were activated by BLM. It also suppressed apoptosis by serum deprivation, which was independent of caspase-3 activation. Therefore, the antiapoptotic neuroprotection by estradiol is mediated by transcription through AP-1 site downstream from JNK and caspase-3 activation. Furthermore, 17alpha estradiol, a stereoisomer without female hormone activity, also provided an antiapoptotic effect. Therefore, the antiapoptotic effect is independent of female hormone activity. PMID- 10834943 TI - The pro-oxidative activity of SOD and nitroxide SOD mimics. AB - Native Cu,Zn-SOD and synthetic SOD mimics sometimes demonstrate an apparently anomalous bell-shaped dose-response relationship when protecting various biological systems from oxidative stress. Several mechanisms have been proposed to account for such an effect, including: overproduction of H(2)O(2), peroxidative activity of SOD, and opposing roles played by O(2)(*-) in both initiation and termination of radical chain reactions. In the present study, ferrocyanide and thiols, which are susceptible to one-electron and two-electron oxidation, respectively, were subjected to a flux of superoxide in the presence and absence of SOD or SOD mimics. The results show that 1) either O(2)(* )/HO(2)(*) or H(2)O(2) alone partially inactivates papain, whereas when combined they act synergistically; 2) nitroxide SOD mimics, but not SOD, exhibit a bell shaped dose-response relationship in protecting papain from inactivation; 3) SOD, which at low dose inhibits superoxide-induced oxidation of ferrocyanide, loses its antioxidative effect as its concentration increases. These findings offer an additional explanation for the pro-oxidative activity of SOD and SOD mimics without invoking any dual activity of O(2)(*-) or a combined effect of SOD and H(2)O(2). The most significant outcome of an increase in SOD level is a decrease of [O(2)(*-)](steady state), rather than any notable elevation of [H(2)O(2)](steady state). As a result, the reaction kinetics of the high oxidation state of each catalyst is altered. In the presence of ultra-low [O(2)(* )](steady state), the oxidized form of SOD [Cu(II),Zn-SOD] or SOD mimic (oxo ammonium cation) does not react with O(2)(*-) but rather oxidizes the target molecule that it was supposed to have protected. Consequently, these catalysts exert an anti- or pro-oxidative effect depending on their concentration. PMID- 10834944 TI - RARbeta involvement in enhancement of lung tumor cell immunogenicity revealed by array analysis. AB - The retinoid receptors (RARs and RXRs) are mediators of the multiple effects of retinoic acid. Of these, the retinoic acid receptor beta2 (RARbeta2) has frequently been shown to be the principal mediator of the growth and tumor suppressive effects of retinoic acid; this gene is inactivated in many epithelial tumors and their derived cell lines. We have searched for genes that are regulated by this isoform and are potentially involved in tumor suppression. Using the Atlas human cDNA array I, we identified 27 genes (not counting RARbeta itself) that are regulated, directly or indirectly, by RARbeta2 when it is transfected into Calu-1, a lung tumor-derived line that does not normally express RARbeta. Several of the affected genes code for proteins whose functions would augment the process of apoptosis and/or the host's immune response. The latter group included ICAM-1 and MHC class I heavy chain, whose protein products play particularly important roles in the mounting of an effective anti-tumor response. We then confirmed by flow cytometry that the observed increases in message levels were reflected in increased cell surface protein levels for ICAM-1 and MHC class I in RARbeta2 transfectants of two RARbeta-deficient lines, Calu-1 and the epidermoid lung cancer-derived line SK-MES. Finally, we showed that RARbeta2 transfection of Calu-1 cells enhanced the heterologous CTL response in both the induction and the effector phases by up to threefold. These results support the hypothesis that down-regulation of these genes (and possibly others) in RARbeta deficient tumor cells contributes to immune system evasion, and suggest a novel therapeutic approach for this disease. PMID- 10834945 TI - Fresh and globular amyloid beta protein (1-42) induces rapid cellular degeneration: evidence for AbetaP channel-mediated cellular toxicity. AB - Amyloid beta peptides (AbetaP) deposit as plaques in vascular and parenchymal areas of Alzheimer's disease (AD) tissues and Down's syndrome patients. Although neuronal toxicity is a feature of late stages of AD, vascular pathology appears to be a feature of all stages of AD. Globular and nonfibrillar AbetaPs are continuously released during normal cellular metabolism, form calcium-permeable channels, and alter cellular calcium level. We used atomic force microscopy, laser confocal microscopy, and calcium imaging to examine the real-time and acute effects of fresh and globular AbetaP(1-42), AbetaP(1-40), and AbetaP(25-35) on cultured endothelial cells. AbetaPs induced morphological changes that were observed within minutes after AbetaP treatment and led to eventual cellular degeneration. Cellular morphological changes were most sensitive to AbetaP(1-42). AbetaP(1-42)-induced morphological changes were observed at nanomolar concentrations and were accompanied by an elevated cellular calcium level. Morphological changes were prevented by anti-AbetaP antibody, AbetaP-channel antagonist zinc, and the removal of extracellular calcium, but not by tachykinin neuropeptide, voltage-sensitive calcium channel blocker cadmium, or antioxidants DTT and Trolox. Thus, nanomolar fresh and globular AbetaP(1-42) induces rapid cellular degeneration by elevating intracellular calcium, most likely via calcium permeable AbetaP channels and not by its interaction with membrane receptors or by activating oxidative pathways. Such rapid degeneration also suggests that the plaques, and especially fibrillar AbetaPs, may not have a direct causative role in AD pathogenic cascades. PMID- 10834946 TI - Fresh and nonfibrillar amyloid beta protein(1-40) induces rapid cellular degeneration in aged human fibroblasts: evidence for AbetaP-channel-mediated cellular toxicity. AB - Alzheimer's disease (AD) is primarily nonfamilial or sporadic (SAD) in origin, although several genetic linkages are reported. Tissues from AD patients contain fibrillar plaques made of 39 to 43 amino acid-long amyloid beta peptide (AbetaP), although the mechanisms of AbetaP toxicity are poorly understood. AbetaP(1-40) is the most prevalent AbetaP present in the neuronal and non-neuronal tissues from SAD patients. AbetaP(1-40) toxicity has been examined mainly after prolonged incubation and correlates with the age and fibrillar morphology of AbetaP(1-40). Globular and nonfibrillar AbetaPs are released continually during normal cellular metabolism; they elevate cellular Ca(2+) and form cation-permeable channels. However, their role in cellular toxicity is poorly understood. We have used an integrated atomic force and light fluorescence microscopy (AFM-LFM), laser confocal microscopy, and calcium imaging to examine real-time and acute effect of fresh and globular AbetaP(1-40) on cultured, aged human, AD-free fibroblasts. AFM images show that freshly prepared AbetaP(1-40) in phosphate-buffered saline (PBS) are globular and do not form fiber for an extended time period. AbetaP(1-40) induced rapid structural modifications, including cytoskeletal reorganization, retraction of cellular processes, and loss of cell-cell contacts, within minutes of incubation. This led to eventual cellular degeneration. AbetaP(1-40)-induced degeneration was prevented by anti-AbetaP antibody, zinc, and Tris, but not by tachykinin neuropeptides. In Ca(2+)-free extracellular medium, AbetaP(1-40) did not induce cellular degeneration. In the presence of extracellular Ca(2+), AbetaP(1-40) induced a sustained increase in the cellular Ca(2+). Thus, short term and acute AbetaP(1-40) toxicity is mediated by Ca(2+) uptake, most likely via calcium-permeable AbetaP pores. Such rapid degeneration does not require fibrillar plaques, suggesting that the plaques may not have any causative role. PMID- 10834947 TI - Protein aging hypothesis of Alzheimer disease. AB - Alzheimer disease (AD), the most common form of aging-related neurodegenerative disorders, is associated with formation of fibrillar deposits of amyloid beta protein (Abeta). While the direct involvement of Abeta in AD has been well documented, the relations between Abeta production, amyloid formation, and neurodegeneration remain unknown. We propose that AD is initiated by a protein aging-related structural transformation in soluble Abeta. We hypothesize that spontaneous chemical modification of aspartyl residues in Abeta to transient succinimide induces a non-native conformation in a fraction of soluble Abeta, rendering it amyloidogenic and neurotoxic. Conformationally altered Abeta is characterized by increased stability in solution and the presence of a non-native beta-turn that determines folding of Abeta in solution and the structure of Abeta subunits incorporated into amyloid fibrils. While the soluble 'non-native' Abeta is both the factor triggering the neurodegenerative cascade and the precursor of amyloid plaques, these two events result from interaction of Abeta with different sets of cellular components and need not coincide in space and time. Extensive literature data and experimental evidence are provided in support of this hypothesis. PMID- 10834948 TI - Discrepant analysis: how can we test a test? PMID- 10834950 TI - Comparison of three commercial rapid identification systems for the unusual gram positive cocci Dolosigranulum pigrum, Ignavigranum ruoffiae, and Facklamia species. AB - We evaluated three rapid identification systems-The Biomerieux rapid ID 32 STREP (ID32), the BBL Crystal rapid gram-positive identification (Crystal), and the Remel IDS RapID STR (IDS) systems-for their ability to identify 7 strains of Alloiococcus otitidis, 27 strains of Dolosigranulum pigrum, 3 strains of Ignavigranum ruoffiae, and 18 strains of 4 different Facklamia species. Since none of these six species of gram-positive cocci are included in the identification databases for these systems, the correct identification for the strains tested should be "unacceptable ID" for the ID32 and Crystal systems or "no choice" for the IDS system. The ID32 system identified all 27 strains of D. pigrum, 6 of 18 Facklamia species, and 2 of 3 cultures of I. ruoffiae as "unacceptable ID." The Crystal system identified 10 of 27 D. pigrum, 2 of 18 Facklamia species, and 2 of 3 I. ruoffiae strains as "unacceptable ID." The IDS system identified only 1 culture of D. pigrum as "no choice," but it also identified 2 cultures of D. pigrum as a "questionable microcode" and 19 cultures of D. pigrum as an "inadequate ID, E. faecalis 90%, S. intermedius 9%." A total of 2 of the 18 cultures of Facklamia and all 3 of the I. ruoffiae cultures were correctly identified as "no choice." The most common misidentifications of Facklamia species by the ID32 and IDS systems were as various Streptococcus species and as Gemella species. In the Crystal system, the most common erroneous identification was Micrococcus luteus. These data indicate the need for the commercial manufacturers of these products to update their databases to include newly described species of gram-positive cocci. PMID- 10834949 TI - Quantitative molecular analysis of virus expression and replication. PMID- 10834951 TI - An analytical model applied to a multicenter pneumococcal enzyme-linked immunosorbent assay study. AB - Pneumococcal conjugate vaccines will eventually be licensed after favorable results from phase III efficacy trials. After licensure of a conjugate vaccine for invasive pneumococcal disease in infants, new conjugate vaccines will likely be licensed primarily on the basis of immunogenicity data rather than clinical efficacy. Analytical methods must therefore be developed, evaluated, and validated to compare immunogenicity results accurately within and between laboratories for different vaccines. At present no analytical technique is uniformly accepted and used in vaccine evaluation studies to determine the acceptable level of agreement between a laboratory result and the assigned value for a given serum sample. This multicenter study describes the magnitude of agreement among 12 laboratories quantifying an identical series of 48 pneumococcal serum specimens from 24 individuals (quality-control sera) by a consensus immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA) developed for this study. After provisional or trial antibody concentrations were assigned to the quality-control serum samples for this study, four methods for comparison of a series of laboratory-determined values with the assigned concentrations were evaluated. The percent error between assigned values and laboratory-determined concentrations proved to be the most informative of the four methods. We present guidelines that a laboratory may follow to analyze a series of quality-control sera to determine if it can reproduce the assigned antibody concentrations within an acceptable level of tolerance. While this study focused on a pneumococcal IgG ELISA, the methods that we describe are easily generalizable to other immunological assays. PMID- 10834952 TI - Rapid detection of mecA-positive and mecA-negative coagulase-negative staphylococci by an anti-penicillin binding protein 2a slide latex agglutination test. AB - A rapid slide latex agglutination (LA) test, MRSA-Screen (Denka Seiken Co., Niigata, Japan), which detects PBP 2a, was tested for its ability to differentiate between mecA-positive and -negative coagulase-negative staphylococci. A total of 463 isolates from 13 species were included in the study. The mecA gene was detected by PCR, and the oxacillin MIC was determined by the agar dilution method according to the guidelines of the National Committee for Clinical Laboratory Standards (NCCLS). The LA test was performed with oxacillin-induced isolates. The true-positive and true-negative results were defined on the basis of the presence or the absence of the mecA gene. By PCR, 251 isolates were mecA positive and 212 were mecA negative. The sensitivities, specificities, and positive and negative predictive values for the LA test compared to the NCCLS breakpoint for oxacillin resistance (>/=0.5 mg/liter) were as follows: for the LA test, 100, 99.5, 99.6, and 100%, respectively; for the NCCLS breakpoint, 100, 60.8, 75.1, and 100%, respectively. One hundred twenty five mecA-positive isolates were also tested by the LA test without induction of PBP 2a; only 72 (57.6%) gave a positive result and required 3 to 15 min for reaction. With induction, all 251 isolates were positive within 3 min. The LA test was reliable in classifying mecA-negative isolates, but it classified isolates for which the oxacillin MIC was >/=0.5 mg/liter as oxacillin susceptible. For the reliable detection of oxacillin resistance by the MRSA Screen in coagulase-negative staphylococci, induction of the mecA gene appears to be necessary. PMID- 10834953 TI - PCR and restriction endonuclease analysis for rapid identification of human adenovirus subgenera. AB - Subgenus identification of adenoviruses is of clinical importance and is as informative as identification by serotype in most clinical situations. A PCR based identification of adenovirus subgenera A, B, C, D, E, and F and sometimes serotypes is described. The PCR uses nonnested primer pair ADRJC1-ADRJC2, which targets a highly conserved region of the adenovirus hexon gene, has a sensitivity of 10 to 40 copies of adenovirus type 2 (Ad2) DNA, and generates 140-bp PCR products from adenovirus serotypes representative of all the subgroups. The PCR products of all subgroups can be differentiated on the basis of the restriction fragment patterns produced by a total of five restriction endonucleases. In addition, serotypes Ad40 and Ad41 (subgroup F) and important serotypes of subgroup D (Ad8, Ad10, Ad19, and Ad37) can easily be differentiated, but serotypes within subgroups B and C cannot. The method was assessed by blind subgenus identification of 56 miscellaneous clinical isolates of adenoviruses. The identities of these isolates at the subgenus level by the PCR correlated 91% (51 of 56) with the results of serotyping by the neutralization test, and 9% (5 of 56) of clinical isolates produced discordant results. PMID- 10834954 TI - Isolation in endothelial cell cultures of chlamydia trachomatis LGV (Serovar L2) from a lymph node of a patient with suspected cat scratch disease. AB - An inguinal lymph node, removed from a 21-year-old Romanian man suspected of having cat scratch disease, was sent to our laboratory for Bartonella culture. Lymph node specimens were inoculated on blood-enriched agar and in an endothelial cell culture system using the centrifugation shell vial technique. Bacteria were grown in cell monolayers and detected as positive with an anti-Bartonella henselae rabbit serum. However, such bacteria were identified as Chlamydia trachomatis biovar LGV serovar L2 by PCR sequencing techniques. Pathological examination of tissue biopsies was compatible with either lymphogranuloma venereum or cat scratch disease. The shell vial system is suitable for isolation of intracellular pathogens responsible for chronic lymphadenopathies, including C. trachomatis, Bartonella species, Francisella tularensis, and mycobacteria. However, care should be taken when identifying Chlamydia spp. and Bartonella spp. using polyclonal antibodies, since species of both genera have common antigens which are responsible for cross-reactions. PMID- 10834955 TI - Monoclonal antibodies directed against conserved epitopes on the nucleocapsid protein and the major envelope glycoprotein of equine arteritis virus. AB - We recently developed a highly effective immunization procedure for the generation of monoclonal antibodies (MAbs) directed against the porcine reproductive and respiratory syndrome virus (E. Weiland, M. Wieczorek-Krohmer, D. Kohl, K. K. Conzelmann, and F. Weiland, Vet. Microbiol. 66:171-186, 1999). The same method was used to produce a panel of 16 MAbs specific for the equine arteritis virus (EAV). Ten MAbs were directed against the EAV nucleocapsid (N) protein, and five MAbs recognized the major viral envelope glycoprotein (G(L)). Two of the EAV G(L)-specific MAbs and one antibody of unknown specificity neutralized virus infectivity. A comparison of the reactivities of the MAbs with 1 U.S. and 22 newly obtained European field isolates of EAV demonstrated that all N-specific MAbs, the three nonneutralizing anti-G(L) MAbs, and the weakest neutralizing MAb (MAb E7/d15-c9) recognized conserved epitopes. In contrast, the two MAbs with the highest neutralization titers bound to 17 of 23 (MAb E6/A3) and 10 of 23 (MAb E7/d15-c1) of the field isolates. Ten of the virus isolates reacted with only one of these two MAbs, indicating that they recognized different epitopes. The G(L)-specific MAbs and the strongly neutralizing MAb of unknown specificity (MAb E6/A3) were used for the selection of neutralization-resistant (NR) virus variants. The observation that the E6/A3-specific NR virus variants were neutralized by MAb E7/d15-c1 and that MAb E6/A3 blocked the infectivity of the E7/d15-c1-specific NR escape mutant confirmed that these antibodies reacted with distinct antigenic sites. Immunoelectron microscopy revealed for the first time that the antigenic determinants recognized by the anti-G(L) MAbs were localized on the virion surface. Surprisingly, although the immunofluorescence signal obtained with the neutralizing antibodies was relatively weak, they mediated binding of about three times as much gold granules to the viral envelope than the nonneutralizing anti-G(L) MAbs. PMID- 10834956 TI - Use of PCR with universal primers and restriction endonuclease digestions for detection and identification of common bacterial pathogens in cerebrospinal fluid. AB - We have designed a universal PCR capable of amplifying a portion of the 16S rRNA gene of eubacteria, including Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus pyogenes, Streptococcus agalactiae, Streptococcus pneumoniae, Enterococcus faecium, Enterococcus faecalis, Mycobacterium tuberculosis, Legionella pneumophila, Escherichia coli, Klebsiella pneumoniae, Serratia marcescens, Enterobacter cloacae, Pseudomonas aeruginosa, Acinetobacter baumannii, Proteus mirabilis, Haemophilus influenzae, and Neisseria meningitidis. The sizes of the amplified products from various bacteria were the same (996 bp), but the restriction patterns of most PCR products generated by HaeIII digestion were different. PCR products from S. aureus and S. epidermidis could not be digested by HaeIII but yielded different patterns when they were digested with MnlI. PCR products from S. pneumoniae, E. faecium, and E. faecalis yielded the same HaeIII digestion pattern but could be differentiated by AluI digestion. PCR products from E. coli, K. pneumoniae, S. marcescens, and E. cloacae also had the same HaeIII digestion pattern but had different patterns when digested with DdeI or BstBI. This universal PCR could detect as few as 10 E. coli or 250 S. aureus organisms. Compared with culture, the sensitivity of this universal PCR for detection and identification of bacteria directly from 150 cerebrospinal fluids was 92.3%. These results suggest that this universal PCR coupled with restriction enzyme analysis can be used to detect and identify bacterial pathogens in clinical specimens. PMID- 10834957 TI - False molecular clusters due to nonrandom association of IS6110 with Mycobacterium tuberculosis. AB - We sought to determine whether nonrandom association of IS6110 with Mycobacterium tuberculosis could result in false-positive clustering in unselected collections of isolates. We typed 196 strains of M. tuberculosis from an unselected community based study in northern Tanzania by IS6110 and polymorphic GC-rich repetitive sequence (PGRS) methodologies. The strains were analyzed by Gelcompar computer software. Analysis of 13 out of 25 groups showed that isolates with identical IS6110 and PGRS patterns were likely to be the same strain. Some IS6110 groups containing strains with identical PGRS patterns had similar IS6110 patterns that differed only by movement of the element. Isolates assigned to a single group (i.e., group 11) on the basis of sharing an identical IS6110 fingerprint pattern did not share identical PGRS fingerprint patterns. Six out of the nine bands in these isolates were in hot-spot locations, as previously defined. This indicates that nonrandom association may result in false-positive clustering in unselected community-based studies. Only strains with identical PGRS and IS6110 patterns are likely to be recently transmitted. PMID- 10834958 TI - Persistence of human papillomavirus DNA in benign and (pre)malignant skin lesions from renal transplant recipients. AB - An extremely diverse group of human papillomavirus (HPV) types consisting of epidermodysplasia verruciformis (EV)-associated HPV types and other cutaneous HPV types (e.g., HPV types 2 and 3) is associated with nonmelanoma cancers and benign lesions of the skin. The frequent presence of multiple HPV types in single skin biopsy specimens of renal transplant recipients prompted us to develop PCR techniques for the detection of distinct (sub)groups of genotypically related cutaneous HPV types, i.e., three subgroups of EV-associated HPV types and two groups (A2 and A4) of other cutaneous HPV types. This approach generally allowed a reliable identification of HPV genotypes by direct sequencing of the PCR products, despite the frequent occurrence of multiple infections. The targeted spectrum of HPV types comprises 66 cutaneous HPV types including 21 putative novel HPV types. We also detected 17 putative novel HPV subtypes. We demonstrated that the skin of nearly all renal transplant recipients who developed various benign and (pre)malignant skin lesions was persistently infected with one or more EV-associated HPV types and/or HPV types belonging to groups A2 and A4. The frequency and distribution of EV-associated HPV and HPV types belonging to groups A2 and A4 were similar in biopsy specimens from hyperkeratotic papillomas (77.5%), squamous cell carcinomas (77. 8%), and actinic keratoses (67.9%) but appeared to be lower in specimens of basal cell carcinomas (35.7%), benign lesions (38.5%), and clinically normal skin (32.3%). These findings suggest that renal transplant recipients are prone to persistent cutaneous HPV infection. Our data do not support the existence of high-risk cutaneous HPV types. PMID- 10834960 TI - High rate of tetracycline resistance in Streptococcus pyogenes in Iran: an epidemiological study. AB - Streptococcus pyogenes, a major human pathogen, is still considered susceptible to beta-lactams, but for other relevant antibiotics, highly variable resistance rates have been reported. Since no data were available from Iran, we tested 1,335 throat isolates from two different regions of the country for their antibiotic susceptibilities and, for comparison, a collection of 80 strains isolated from 1989 to 1991. Erythromycin resistance was uncommon (0.6%), whereas an overall high rate of tetracycline resistance was found, increasing between 1989-1991 and 1995-1997 from 23 to 42%. The tetracycline-resistant strains belonged to more than 10 different T types, the majority being types 4, 11, and B3264. By conventional M typing of 406 tetracycline-resistant isolates, more than 20 different M types were found. Approximately 50% of the strains were nontypeable by T agglutination as well as serological M typing; however, by genotyping by a combined PCR-capture-enzyme-linked immunosorbent assay, many of these strains were successfully emm typed. We conclude that the high rate of tetracycline resistance among Iranian S. pyogenes isolates is due to multiclonal dissemination of resistance within the streptococcal population rather than epidemic spread of single clones. PMID- 10834959 TI - A European multicenter study of immunoblotting in serodiagnosis of lyme borreliosis. AB - A European multicenter study of immunoblotting for the serodiagnosis of Lyme borreliosis showed considerable variation in results obtained from tests with a panel of 227 serum samples. Six laboratories used different immunoblot methods, and a wide range of bands was detected in all the assays. Multivariable logistic regression analysis of data from individual laboratories was used to determine the most discriminatory bands for reliable detection of antibodies to Borrelia burgdorferi sensu lato. These bands were used to construct individual interpretation rules for the immunoblots used in the six laboratories. Further analysis identified a subset of eight bands, which were important in all the laboratories, although with variations in significance. Possible European rules, all closely related, were formulated from these bands, although there was no single rule that gave high levels of sensitivity and specificity for all the laboratories. This is a reflection of the wide range of methodologies used, especially the use of different species and strains of B. burgdorferi sensu lato. The panel of European rules provides a framework for immunoblot interpretation which may be adapted in relation to the characteristics of Lyme borreliosis in local areas. PMID- 10834961 TI - Evaluation of a new system, VITEK 2, for identification and antimicrobial susceptibility testing of enterococci. AB - We evaluated the new automated VITEK 2 system (bioMerieux) for the identification and antimicrobial susceptibility testing of enterococci. The results obtained with the VITEK 2 system were compared to those obtained by reference methods: standard identification by the scheme of Facklam and Sahm [R. R. Facklam and D. F. Sahm, p. 308-314, in P. R. Murray et al., ed., Manual of Clinical Microbiology, 6th ed., 1995] and with the API 20 STREP system and, for antimicrobial susceptibility testing, broth microdilution and agar dilution methods by the procedures of the National Committee for Clinical Laboratory Standards. The presence of vanA and vanB genes was determined by PCR. A total of 150 clinical isolates were studied, corresponding to 60 Enterococcus faecalis, 55 Enterococcus faecium, 26 Enterococcus gallinarum, 5 Enterococcus avium, 2 Enterococcus durans, and 2 Enterococcus raffinosus isolates. Among those isolates, 131 (87%) were correctly identified to the species level with the VITEK 2 system. Approximately half of the misidentifications were for E. faecium with low-level resistance to vancomycin, identified as E. gallinarum or E. casseliflavus; however, a motility test solved the discrepancies and increased the agreement to 94%. Among the strains studied, 66% were vancomycin resistant (57 VanA, 16 VanB, and 26 VanC strains), 23% were ampicillin resistant (MICs, >/=16 microgram/ml), 31% were high-level gentamicin resistant, and 45% were high level streptomycin resistant. Percentages of agreement for susceptibility and resistance to ampicillin, vancomycin, and teicoplanin and for high-level gentamicin resistance and high-level streptomycin resistance were 93, 95, 97, 97, and 96%, respectively. The accuracy of identification and antimicrobial susceptibility testing of enterococci with the VITEK 2 system, together with the significant reduction in handling time, will have a positive impact on the work flow of the clinical microbiology laboratory. PMID- 10834962 TI - Isolation and characterization of glycopeptide-resistant enterococci from hospitalized patients over a 30-month period. AB - In February 1996, a Hospital Infection Control Practices Advisory Committee-style screening program was commenced to isolate and subsequently characterize glycopeptide-resistant enterococci (GRE) from patients at a hospital trust in Glasgow, Scotland. Over the next 30 months, GRE were isolated from 154 patients. GRE were isolated from patients in traditionally high-risk areas such as the renal unit and intensive care unit and also in areas considered to be lower risk, including medical wards and associated long-stay geriatric hospitals. The majority (90%) of isolates were Enterococcus faecium vanB. The remaining isolates consisted of seven E. faecalis (vanA), three E. gallinarum (vanC), and a further six E. faecium (five vanA, one both vanA and vanB) isolates. Analysis of SmaI digested DNA by pulsed-field gel electrophoresis revealed that 34 of 40 (85%) VanB E. faecium isolates were identical or closely related, while 11 of 13 (85%) VanA GRE were distinct. High-level aminoglycoside resistance was seen in less than 8% of isolates. VanB E. faecium isolates were almost uniformly resistant to ampicillin and tetracycline. In this study, GRE have been isolated over a prolonged period from a broad range of patients. Glycopeptide resistance within the study hospital trust appeared to be mainly due to the clonal dissemination of a single strain of E. faecium VanB. PMID- 10834963 TI - Accuracy and precision of quantitative calibrated loops in transfer of bronchoalveolar lavage fluid. AB - Quantitative cultures of bronchoalveolar lavage (BAL) fluid are important in the diagnosis of ventilator-associated pneumonia, and calibrated loops are commonly used to set up these cultures. In this study, the performances of calibrated 0.010- and 0.001-ml loops in the transfer of BAL fluid were determined. Five loops of one lot from seven manufacturers were tested. Calibrations were performed by the gravimetric method (0.010-ml loops) and the colorimetric method (0.001-ml loops). Most of the 0.010-ml loops displayed a precision that was less than 10%, but six of them showed very poor accuracies as they transferred a deficiency (nichrome loops) or an excess (disposable loops) of BAL fluid that exceeded +/-10%. The mean maximum and minimum BAL fluid volumes delivered by the 0.010-ml loops differed by a factor 3. The 0.001-ml loops displayed acceptable precision. Five of them showed inaccuracies of /=1:40 would be called HHV-8 positive. A negative or low titer would be called HHV-8 negative. If a population has a high percentage of persons who test positive by the combined peptide EIAs, then a MIFA could be performed with the negative specimens to determine if any positive specimens are being missed. Alternatively, if a population has a low percentage that test positive, then a MIFA could be performed with a subset of the negative specimens for the same reason. As described above, only a titer of >/=1:40 would be considered HHV-8 positive. PMID- 10834973 TI - Frequency of low-level bacteremia in children from birth to fifteen years of age. AB - A single blood culture inoculated with a small volume of blood is still frequently being used for the diagnosis of bacteremia in children because of the continued belief by many that bacteria are usually found in high concentrations in the blood of pediatric patients with sepsis. To determine the importance of both blood volume cultured and the number of culture devices required for the reliable detection of pathogens in our pediatric population, blood from children from birth to 15 years of age and with suspected bacteremia at York Hospital (a 500-bed community hospital) was inoculated into at least a Pediatric Isolator (Wampole Laboratories; 1.5 ml of blood) or a standard Isolator (10 ml of blood) and a bottle of ESP anaerobic broth (Trek Diagnostic Systems; 0.5 to 10 ml of blood). The use of a second Isolator and additional aerobic and anaerobic bottles and the total blood volume recommended for cultures (2 to 60 ml) depended on the weight and total blood volume of each patient. One hundred forty-seven pathogens were recovered from the blood of 137 (3.6%) of 3,829 children for whom culturing was done. Of 121 septic episodes for which the concentration of pathogens in the blood could be determined using Isolators, 73 (60. 3%) represented low-level bacteremia (/=10(5) bacteria (bacterium/cell ratio = 0.25). The cagA-positive, cytotoxin-producing strains (HP64, HP57, and HP87) caused significantly stronger inhibition of intracellular mucin synthesis than the cagA-positive, non-cytotoxin-producing strains (HP05, HP83, and HP84). The cagA-negative, non-cytotoxin-producing strains (HP01, HP04, and HP85) did not affect intracellular mucin synthesis. The results indicate that H. pylori directly impairs mucin synthesis in gastric mucous cells and that cytotoxic cagA-positive strains cause more profound inhibition of mucin synthesis. We suggest that the increased inhibitory effect of cagA-positive, cytotoxin-producing strains on mucin synthesis can be considered one possible factor responsible for the increased risk of developing peptic ulceration with these H. pylori strains. PMID- 10834980 TI - Oral colonization, phenotypic, and genotypic profiles of Candida species in irradiated, dentate, xerostomic nasopharyngeal carcinoma survivors. AB - The aim of this study was to investigate oral yeast colonization and oral yeast strain diversity in irradiated (head and neck), dentate, xerostomic individuals. Subjects were recruited from a nasopharyngeal carcinoma clinic and were segregated into group A (age, <60 years [n = 25; average age +/- standard deviation (SD), 48 +/- 6 years; average postirradiation time +/- SD, 5 +/- 5 years]) and group B (age, >/=60 years [n = 8; average age +/- SD, 67 +/- 4 years; average postirradiation time +/- SD, 2 +/- 2 years]) and were compared with age- and sex-matched healthy individuals in group C (age, <60 years [n = 20; average age +/- SD, 44 +/- 12 years] and group D (age, >/=60 years [n = 10; average age, 70 +/- 3 years]). Selective culture of oral rinse samples was carried out to isolate, quantify, and speciate yeast recovery. All test subjects underwent a 3 month comprehensive oral and preventive care regimen plus topical antifungal therapy, if indicated. A total of 12 subjects from group A and 5 subjects from group B were recalled for reassessment of yeast colonization. Sequential (pre- and posttherapy) Candida isolate pairs from patients were phenotypically (all isolate pairs; biotyping and resistotyping profiles) and genotypically (Candida albicans isolate pairs only; electrophoretic karyotyping by pulsed-field gel electrophoresis, restriction fragment length polymorphism [RFLP], and randomly amplified polymorphic DNA [RAPD] assays) evaluated. All isolates were Candida species. Irradiated individuals were found to have a significantly increased yeast carriage compared with the controls. The isolation rate of Candida posttherapy remained unchanged. A total of 9 of the 12 subjects in group A and 3 of the 5 subjects in group B harbored the same C. albicans or Candida tropicalis phenotype at recall. Varying degrees of congruence in the molecular profiles were observed when these sequential isolate pairs of C. albicans were analyzed by RFLP and RAPD assays. Variations in the genotype were complementary to those in the phenotypic characteristics for some isolates. In conclusion, irradiation-induced xerostomia seems to favor intraoral colonization of Candida species, particularly C. albicans, which appeared to undergo temporal modifications in clonal profiles both phenotypically and genotypically following hygienic and preventive oral care which included topical antifungal therapy, if indicated. We postulate that the observed ability of Candida species to undergo genetic and phenotypic adaptation could strategically enhance its survival in the human oral cavity, particularly when salivary defenses are impaired. PMID- 10834981 TI - A comparison of seven tests for serological diagnosis of tuberculosis. AB - Seven serological tests, two immunochromatographic tests, ICT Tuberculosis and RAPID TEST TB, and five enzyme-linked immunosorbent assays, TUBERCULOSIS IgA EIA, PATHOZYME-TB complex, PATHOZYME-MYCO IgG, PATHOZYME-MYCO IgA, and PATHOZYME-MYCO IgM, were evaluated simultaneously with 298 serum samples from three groups of individuals: 44 patients with active tuberculosis, 204 controls who had undergone the Mantoux test (89 Mantoux test-positive and 115 Mantoux test-negative controls), and 50 anonymous controls. The sensitivities of the tests with sera from patients with active tuberculosis were poor to modest, ranging from 16 to 57%. All the tests performed equally with sera from subgroups of those with active tuberculosis, those with pulmonary (33 patients) versus extrapulmonary (11 patients) disease, and those who were smear positive (24 patients) versus smear negative (12 patients) (P > 0.05). The specificities of the tests ranged from 80 to 97% with sera from the Mantoux test controls and 62 to 100% with sera from the anonymous controls. The TUBERCULOSIS IgA EIA had the highest sensitivity (57%) with sera from patients with active tuberculosis, with a high specificity of 93% with sera from the Mantoux test controls, but a very poor specificity of 62% with sera from the anonymous controls. Overall, ICT Tuberculosis followed by PATHOZYME MYCO IgG had the best performance characteristics, with sensitivities of 41 and 55%, respectively, with sera from patients with active tuberculosis and specificities of 96 and 89%, respectively, with sera from the Mantoux test controls and 88 and 90%, respectively, with sera from the anonymous controls. By combining all the test results, a maximum sensitivity of 84% was obtained, with reciprocal drops in specificities to 55 and 42% for the Mantoux test controls and anonymous controls, respectively. The best combination was that of ICT Tuberculosis and PATHOZYME-MYCO IgG, with a sensitivity of 66% and a specificity of 86% for the Mantoux test controls and a sensitivity and specificity of 78% for the anonymous controls. While a negative result by any one of these tests would be useful in helping to exclude disease in a population with a low prevalence of tuberculosis, a positive result may aid in clinical decision making when applied to symptomatic patients being evaluated for active tuberculosis. PMID- 10834982 TI - Evaluation of immunoglobulin M (IgM) and IgG enzyme immunoassays in serologic diagnosis of West Nile Virus infection. AB - A unique urban encephalitis epidemic in Romania signaled the emergence of neurological infection due to West Nile (WN) virus as a novel public health threat in Eastern Europe and provided an opportunity to evaluate patterns of immunoglobulin G (IgG) and IgM reactivity in IgM capture and IgG enzyme-linked immunosorbent assays (ELISAs). WN virus infection was diagnosed serologically in 236 of 290 patients from whom acute serum or cerebrospinal fluid (CSF) samples were available. In 37% of serum samples and in 25% of CSF samples collected in the first week of illness, anti-WN virus IgM antibody was detected in the absence of virus-specific IgG. The switch to an IgG antibody response occurred after 4 to 5 days of illness and earlier in CSF than in serum. A specific humoral immune response was detected in the CSF before the serum in some patients for whom paired CSF and serum samples from the same day were available. IgM antibody in convalescent serum samples persisted beyond 2 months after the onset of illness in more than 50% of patients. ELISA optical density values and antibody concentrations were well correlated for both IgM and IgG immunoassays. Anti-WN virus IgM antibody in acute-phase samples did not cross-react significantly with flaviviruses in other antigenic groups. PMID- 10834983 TI - Cloning, characterization, and expression of a 200-kilodalton diagnostic antigen of Babesia bigemina. AB - Current serological tests for Babesia bigemina use semipurified merozoite antigens derived from infected erythrocytes. One of the major drawbacks of these tests is that antigen quality can vary from batch to batch. Since the quality of the antigen contributes to the sensitivity and specificity of serological tests, the use of standardized recombinant antigens should ensure consistency in assay quality. Previously, a 200-kDa merozoite antigen (p200) was identified as a candidate diagnostic antigen for use in a serological assay for the detection of B. bigemina antibodies in infected cattle. In this study, we have cloned, characterized, and expressed p200. A 3.5-kbp cDNA clone encoding p200 was isolated and shown to be almost full length, lacking approximately 300 bp at the 5' end. The predicted amino acid sequence shows that p200 consists of a long, highly charged central repeat region of an uninterrupted alpha helix, indicative of a fibrous protein. Immunoelectron microscopy localized p200 to the merozoite cytoplasm, suggesting that the antigen may be a structural protein involved in forming filament structures within the cytoskeleton. The 3.5-kbp cDNA was expressed in bacteria as a fusion protein with glutathione S-transferase (GST), but the yield was poor. To improve the yield, cDNA fragments encoding antigenic domains of p200 were expressed as fusions with GST. One of these fusion proteins, C1A-GST, is composed of a 7-kDa fragment of the p200 repeat region and contains epitopes that react strongly with sera from cattle experimentally infected with B. bigemina. Recombinant C1A-GST should permit the development of an improved enzyme-linked immunosorbent assay for the detection of antibodies against B. bigemina. PMID- 10834984 TI - Cloning and sequencing of a part of the heat shock protein 65 gene (hsp65) of "Tropheryma whippelii" and its use for detection of "T. whippelii" in clinical specimens by PCR. AB - Using broad-spectrum primers, we have amplified, cloned, and sequenced a 620-bp fragment of the "Tropheryma whippelii" heat shock protein 65 gene (hsp65) from the heart valve of a patient with Whipple's endocarditis. The deduced amino acid sequence shows high similarity to those from actinobacteria, confirming that "T. whippelii" is indeed a member of this phylum. Based on the nucleotide sequence, we have developed a "T. whippelii"-specific seminested PCR. Seventeen patients shown to be positive by 16S ribosomal DNA (rDNA) PCR and/or internal transcribed spacer (ITS) PCR were also positive by hsp65 PCR. All 33 control specimens from patients without Whipple's disease and negative for "T. whippelii" by both seminested 16S rDNA and ITS PCR remained negative. All amplicons digested with XhoI revealed an identical restriction pattern. Eight of the 17 hsp65 amplicons representing all three previously described ITS types were sequenced. Three of the amplicons showed slight differences, but none of the mutations detected affected the amino acid sequence of the corresponding protein. We conclude that the hsp65 gene is a suitable target for the specific detection of "T. whippelii." Its product represents a putative antigen for a future serodiagnostic assay. PMID- 10834985 TI - Comparative evaluation of PASCO and national committee for clinical laboratory standards M27-A broth microdilution methods for antifungal drug susceptibility testing of yeasts. AB - The PASCO antifungal susceptibility test system, developed in collaboration with a commercial company, is a broth microdilution assay which is faster and easier to use than the reference broth microdilution test performed according to the National Committee for Clinical Laboratory Standards (NCCLS) document M27-A guidelines. Advantages of the PASCO system include the system's inclusion of quality-controlled, premade antifungal panels containing 10, twofold serial dilutions of drugs and a one-step inoculation system whereby all wells are simultaneously inoculated in a single step. For the prototype panel, we chose eight antifungal agents for in vitro testing (amphotericin B, flucytosine, fluconazole, ketoconazole, itraconazole, clotrimazole, miconazole, and terconazole) and compared the results with those of the NCCLS method for testing 74 yeast isolates (14 Candida albicans, 10 Candida glabrata, 10 Candida tropicalis, 10 Candida krusei, 10 Candida dubliniensis, 10 Candida parapsilosis, and 10 Cryptococcus neoformans isolates). The overall agreements between the methods were 91% for fluconazole, 89% for amphotericin B and ketoconazole, 85% for itraconazole, 80% for flucytosine, 77% for terconazole, 66% for miconazole, and 53% for clotrimazole. In contrast to the M27-A reference method, the PASCO method classified as resistant seven itraconazole-susceptible isolates (9%), two fluconazole-susceptible isolates (3%), and three flucytosine-susceptible isolates (4%), representing 12 major errors. In addition, it classified two fluconazole resistant isolates (3%) and one flucytosine-resistant isolate (1%) as susceptible, representing three very major errors. Overall, the agreement between the methods was greater than or equal to 80% for four of the seven species tested (C. dubliniensis, C. glabrata, C. krusei, and C. neoformans). The lowest agreement between methods was observed for miconazole and clotrimazole and for C. krusei isolates tested against terconazole. When the data for miconazole and clotrimazole were removed from the analysis, agreement was >/=80% for all seven species tested. Therefore, the PASCO method is a suitable alternative procedure for the testing of the antifungal susceptibilities of the medically important Candida spp. and C. neoformans against a range of antifungal agents with the exceptions only of miconazole and clotrimazole and of terconazole against C. krusei isolates. PMID- 10834986 TI - Misidentifying helicobacters: the Helicobacter cinaedi example. AB - Whole-cell protein electrophoresis and biochemical examination by means of a panel of 64 tests were used to identify 14 putative helicobacters to the species level. The results were confirmed by means of DNA-DNA hybridization experiments and were used to discuss misidentification of helicobacters based on 16S rRNA gene sequence data. The data indicated that comparison of near-complete 16S ribosomal DNA sequences does not always provide conclusive evidence for species level identification and may prove highly misleading. The data also indicated that "Helicobacter westmeadii" is a junior synonym of Helicobacter cinaedi and that Helicobacter sp. strain Mainz belongs to the same species. H. cinaedi occurs in various animal reservoirs, including hamsters, dogs, cats, rats, and foxes. Appropriate growth conditions and identification strategies will be required to establish the genuine significance of this widely distributed Helicobacter species. PMID- 10834987 TI - A survey of stool culturing practices for vibrio species at clinical laboratories in Gulf Coast states. AB - Non-cholera Vibrio infections are an important public health problem. Non-cholera Vibrio species usually cause sporadic infections, often in coastal states, and have also caused several recent nationwide outbreaks of gastroenteritis in the United States. We report a survey of laboratory stool culturing practices for Vibrio among randomly selected clinical laboratories in Gulf Coast states (Alabama, Florida, Louisiana, Mississippi, and Texas). Interviews conducted with the microbiology supervisors of 201 clinical laboratories found that 164 (82%) received stool specimens for culture. Of these, 102 (62%) of 164 processed stool specimens on site, and 20 (20%) of these 102 laboratories cultured all stool specimens for Vibrio, indicating that at least 34,463 (22%) of 152, 797 stool specimens were cultured for Vibrio. This survey suggests that despite an increased incidence of non-cholera Vibrio infections in Gulf Coast states, a low percentage of clinical laboratories routinely screen all stool specimens, and fewer than 25% of stool specimens collected are routinely screened for non cholera Vibrio. PMID- 10834988 TI - Extraction-free, filter-based template preparation for rapid and sensitive PCR detection of pathogenic parasitic protozoa. AB - Within the last several years, the protozoan parasites Cyclospora cayetanensis, Cryptosporidium parvum, and microsporidia have become recognized as important, rapidly emerging human pathogens in immunocompromised and immunocompetent individuals. Since the early 1990s, many of the reported outbreaks of enteric illness caused by these microorganisms have been attributed to food- and water borne contamination. Many inherent obstacles affect the success of current surveillance and detection methods used to monitor and control levels of contamination by these pathogens. Unlike methods that incorporate preenrichment for easier and unambiguous identification of bacterial pathogens, similar methods for the detection of parasitic protozoa either are not currently available or cannot be performed in a timely manner. We have developed an extraction-free, filter-based protocol to prepare DNA templates for use in PCR to identify C. cayetanensis and C. parvum oocysts and microsporidia spores. This method requires only minimal preparation to partially purify and concentrate isolates prior to filter application. DNA template preparation is rapid, efficient, and reproducible. As few as 3 to 10 parasites could be detected by PCR from direct application to the filters. In studies, as few 10 to 50 Encephalitozoon intestinalis spores could be detected when seeded in a 100-microliter stool sample and 10 to 30 C. cayetanensis oocysts could be detected per 100 g of fresh raspberries. This protocol can easily be adapted to detect parasites from a wide variety of food, clinical, and environmental samples and can be used in multiplex PCR applications. PMID- 10834989 TI - Development of antigen detection assay for diagnosis of tuberculosis using sputum samples. AB - The rising incidence of tuberculosis worldwide means an increasing burden on diagnostic facilities, so tests simpler than Ziehl-Neelsen staining are needed. Such tests should be objective, reproducible, and have at least as good a detection limit as 10(4) bacteria/ml. A capture enzyme-linked immunosorbent assay (ELISA) was developed for detection of lipoarabinomannan (LAM) in human sputum samples. As a capture antibody, we used a murine monoclonal antibody against LAM, with rabbit antiserum against Mycobacterium tuberculosis as a source of detector antibodies. The sensitivity of the capture ELISA was evaluated by using purified LAM and M. tuberculosis whole cells. We were able to detect 1 ng of purified LAM/ml and 10(4) M. tuberculosis whole cells/ml. LAM could also be detected in culture filtrate of a 3-week-old culture of M. tuberculosis. The culture filtrate contained approximately 100 microgram of LAM/ml. The detection limit in sputum pretreated with N-acetyl-L-cysteine and proteinase K was 10(4) M. tuberculosis whole cells per ml. Thirty-one (91%) of 34 sputum samples from 18 Vietnamese patients with tuberculosis (32 smear positive and 2 smear negative) were positive in the LAM detection assay. In contrast, none of the 25 sputum samples from 21 nontuberculous patients was positive. This specific and sensitive assay for the detection of LAM in sputum is potentially useful for the diagnosis of tuberculosis. PMID- 10834990 TI - Comparative evaluation of three different genotyping methods for investigation of nosocomial outbreaks of Legionnaires' disease in hospitals. AB - The increased incidence of nosocomial Legionnaires' disease in two hospitals prompted investigation of possible environmental sources. In the search for an effective DNA-typing technique for use in hospital epidemiology, the performance and convenience of three methods-SfiI macrorestriction analysis (MRA), amplified fragment length polymorphism (AFLP), and arbitrarily primed PCR (AP-PCR)-were compared. Twenty-nine outbreak-associated and eight nonassociated strains of Legionella pneumophila with 13 MRA types and subtypes were investigated. These strains comprised isolates from bronchoalveolar lavages, from environmental, patient-related sources, and type strains. All three typing methods detected one predominant genotype associated with the outbreaks in both hospitals. All of them correctly assigned epidemiologically associated, environmental isolates to their respective patient specimens. AP-PCR was the least discriminating and least reproducible technique. In contrast, AFLP was demonstrated as being the method with the best interassay reproducibility (90%) and concordance (94%) in comparison to the genotyping standard of MRA and the epidemiological data. Analysis of AFLP fragments revealed 12 different types and subtypes. Because of its simplicity and reproducibility, AFLP proved to be the most effective technique in outbreak investigation. PMID- 10834991 TI - Urogenital Chlamydia trachomatis serovars in men and women with a symptomatic or asymptomatic infection: an association with clinical manifestations? AB - To determine whether certain Chlamydia trachomatis serovars are preferentially associated with a symptomatic or an asymptomatic course of infection, C. trachomatis serovar distributions were analyzed in symptomatically and asymptomatically infected persons. Furthermore, a possible association between C. trachomatis serovars and specific clinical symptoms was investigated. C. trachomatis-positive urine specimens from 219 asymptomatically infected men and women were obtained from population-based screening programs in Amsterdam. Two hundred twenty-one C. trachomatis-positive cervical and urethral swabs from symptomatically and asymptomatically infected men and women were obtained from several hospital-based departments. Serovars were determined using PCR-based genotyping, i.e., restriction fragment length polymorphism analysis of the nested PCR-amplified omp1 gene. The most prevalent C. trachomatis serovars, D, E, and F, showed no association with either a symptomatic or asymptomatic course of infection. The most prominent differences found were (i) the association of serovar Ga with symptoms in men (P = 0.0027), specifically, dysuria (P < 0.0001), and (ii) detection of serovar Ia more often in asymptomatically infected people (men and women) (P = 0.035). Furthermore, in women, serovar K was associated with vaginal discharge (P = 0.002) and serovar variants were found only in women (P = 0.045). PMID- 10834992 TI - Molecular characterization of Campylobacter jejuni from patients with Guillain Barre and Miller Fisher syndromes. AB - Campylobacter jejuni has been identified as the predominant cause of antecedent infection in Guillain-Barre syndrome (GBS) and Miller Fisher syndrome (MFS). The risk of developing GBS or MFS may be higher after infection with specific C. jejuni types. To investigate the putative clonality, 18 GBS- or MFS-related C. jejuni strains from The Netherlands and Belgium and 17 control strains were analyzed by serotyping (Penner and Lior), restriction fragment length polymorphism analysis of PCR products of the flaA gene, amplified fragment length polymorphism analysis, pulsed-field gel electrophoresis, and randomly amplified polymorphic DNA analysis. Serotyping revealed 10 different O serotypes and 7 different Lior serotypes, thereby indicating a lack of serotype clustering. Two new O serotypes, O:35 and O:13/65, not previously associated with GBS or MFS were found. Serotype O:19 was encountered in 2 of 18 strains, and none was of serotype O:41. The results of all genotypic methods also demonstrated substantial heterogeneity. No clustering of GBS- or MFS-related strains occurred and no molecular marker capable of separating pathogenic GBS or MFS from non-GBS- or non MFS-related enteritis strains could be identified in this study. Sialic-acid containing lipopolysaccharides (LPS) are thought to be involved in the triggering of GBS or MFS through molecular mimicry with gangliosides in human peripheral nerves. Therefore, further characterization of GBS- or MFS-related C. jejuni should target the genes involved in the synthesis of LPS and the incorporation of sialic acid. PMID- 10834993 TI - Identification of medically important yeasts using PCR-based detection of DNA sequence polymorphisms in the internal transcribed spacer 2 region of the rRNA genes. AB - Identification of medically relevant yeasts can be time-consuming and inaccurate with current methods. We evaluated PCR-based detection of sequence polymorphisms in the internal transcribed spacer 2 (ITS2) region of the rRNA genes as a means of fungal identification. Clinical isolates (401), reference strains (6), and type strains (27), representing 34 species of yeasts were examined. The length of PCR-amplified ITS2 region DNA was determined with single-base precision in less than 30 min by using automated capillary electrophoresis. Unique, species specific PCR products ranging from 237 to 429 bp were obtained from 92% of the clinical isolates. The remaining 8%, divided into groups with ITS2 regions which differed by /=99%. Seven clinical isolates contained ITS2 sequences that did not agree with their phenotypic identification, and ITS2-based phylogenetic analyses indicate the possibility of new or clinically unusual species in the Rhodotorula and Candida genera. This work establishes an initial database, validated with over 400 clinical isolates, of ITS2 length and sequence polymorphisms for 34 species of yeasts. We conclude that size and restriction analysis of PCR-amplified ITS2 region DNA is a rapid and reliable method to identify clinically significant yeasts, including potentially new or emerging pathogenic species. PMID- 10834994 TI - Dynamics of meningococcal long-term carriage among university students and their implications for mass vaccination. AB - In the 1997-98 academic year, we conducted a longitudinal study of meningococcal carriage and acquisition among first-year students at Nottingham University, Nottingham, United Kingdom. We examined the dynamics of long-term meningococcal carriage with detailed characterization of the isolates. Pharyngeal swabs were obtained from 2,453 first-year students at the start of the academic year (October), later on during the autumn term, and again in March. Swabs were immediately cultured on selective media, and meningococci were identified and serologically characterized. Nongroupable strains were genetically grouped using a PCR-based assay. Pulsed-field gel electrophoresis was used to determine the link between sequential isolates. Of the carriers initially identified in October, 44.1% (98 of 222) were still positive later on in the autumn (November or December); 57.1% of these remained persistent carriers at 6 months. Of the index carriers who lost carriage during the autumn, 16% were recolonized at 6 months. Of 344 index noncarriers followed up, 22.1% acquired carriage during the autumn term and another 13.7% acquired carriage by March. Overall, 43.9% (397 of 904) of the isolates were noncapsulated (serologically nongroupable); by PCR based genogrouping, a quarter of these belonged to the capsular groups B and C. The ratio of capsulated to noncapsulated forms for group B and C strains was 2.9 and 0.95, respectively. Sequential isolates of persistent carriers revealed that individuals may carry the same or entirely different organisms at different times. We identified three strains that clearly switched their capsular expression on and off at different times in vivo. One student developed invasive meningococcal disease after carrying the same organism for over 7 weeks. The study revealed a high rate of turnover of meningococcal carriage among students. Noncapsulated organisms are capable of switching their capsular expression on and off (both ways) in the nasopharynx, and group C strains are more likely to be noncapsulated than group B strains. Carriage of a particular meningococcal strain does not necessarily protect against colonization or invasion by a homologous or heterologous strain. PMID- 10834995 TI - Intestinal lesions associated with disseminated candidiasis in an experimental animal model. AB - In human patients, disseminated candidiasis, a life-threatening disease for immunocompromised patients, is often associated with intestinal lesions. In this study, we demonstrate that immunosuppressed gnotobiotic (IGB) piglets orally inoculated with wild-type Candida albicans developed extensive intestinal lesions and disseminated infection. Severe ulceration of the ileal mucosa was observed overlying regions of colonization and necrosis of the gut-associated lymphoid tissue. Despite the high susceptibility of IGB piglets to many microbial pathogens, an avirulent mutant strain of C. albicans failed to produce intestinal lesions and exhibited poor dissemination, demonstrating that these effects required virulent organisms. It is likely that in IGB piglets, as in human patients, intestinal lesions provide the mechanism for escape of C. albicans from the gastrointestinal tract. Multinucleated giant cells containing fungal organisms were observed within lymph nodes and lymphatic vessels, and as with other pathogens, such cells could provide a mechanism for dissemination of C. albicans. PMID- 10834997 TI - Recovery of Bordetella holmesii from patients with pertussis-like symptoms: use of pulsed-field gel electrophoresis to characterize circulating strains. AB - A 4-year retrospective study showing that we isolated Bordetella holmesii, but not Bordetella pertussis, from patients with pertussis-like symptoms was performed. From 1995 through 1998, we isolated B. holmesii from 32 nasopharyngeal specimens that had been submitted from patients suspected of having pertussis. Previously, B. holmesii had been associated mainly with septicemia and was not thought to be associated with respiratory illness. A study was undertaken to describe the characteristics of the B. holmesii isolates recovered and why we were successful in detecting the organism in nasopharyngeal specimens. B. holmesii isolates were characterized for drug sensitivities and for genetic relatedness by pulsed-field gel electrophoresis (PFGE). These isolates, an additional strain of B. holmesii isolated from a blood culture and previously confirmed by the Centers for Disease Control and Prevention, Atlanta, Ga., and 14 other clinical isolates of Bordetella spp., including 4 of B. bronchiseptica, 5 of B. parapertussis, and 5 of B. pertussis, were studied. They were all separately inoculated on three Bordet Gengou (BG) selective media containing either 0.625 microgram of oxacillin per ml, 40 microgram of cephalexin per ml, or 2.5 microgram of methicillin per ml, on BG agar with no antibiotic (control), and on charcoal agar (CA) with and without 40 microgram of cephalexin per ml. We found that cephalexin, the antibiotic commonly incorporated in both CA and BG agar for the recovery of Bordetella spp., is inhibitory to the growth of B. holmesii. In addition, the genotypic analysis of the 32 B. holmesii isolates by PFGE following restriction with XbaI and SpeI identified the dominant strains circulating during the study period. PMID- 10834998 TI - Comparison of a new colorimetric assay with the NCCLS broth microdilution method (M-27A) for antifungal drug MIC determination. AB - We evaluated a new microtiter assay for antifungal susceptibility testing based on a colorimetric reaction to monitor fungal substrate utilization. This new method (rapid susceptibility assay [RSA]) provides quantitative endpoint readings in less than 8 h compared with visual determination of MIC by the National Committee for Clinical Laboratory Standards (NCCLS) broth microdilution method, which requires a minimum of 48 h of incubation. In this study, we tested clinical isolates from each of the following species: Candida albicans (20 isolates), C. glabrata (20 isolates), C. krusei (19 isolates), C. tropicalis (19 isolates), and C. parapsilosis (28 isolates). RSA and NCCLS broth dilution methods were used to determine the MICs of amphotericin B, fluconazole, itraconazole, and 5 flucytosine for all 106 isolates. RPMI 1640 medium buffered with morpholinopropanesulfonic acid was used for both methods; however, glucose and inoculum concentrations in the RSA were modified. RSA MICs were determined as the lowest drug concentration that prevented glucose consumption by the organism after 6 h of incubation. MICs obtained from the RSA were compared with those obtained from the NCCLS M-27A method read at 24 and 48 h. MIC pairs were considered in agreement when the difference between the pairs was within 2 twofold dilutions. For the 106 isolates tested, amphotericin B and 5-flucytosine demonstrated the highest agreement in MICs between the two methods (100 and 98%, respectively), whereas fluconazole and itraconazole produced less favorable MIC agreement (63.2 and 61.3%, respectively). The azole MIC differences between the two methods were significantly reduced when lower inocula were used with a prolonged incubation time. This preliminary comparison suggests that this rapid procedure may be a reliable tool for the in vitro determination of MICs of amphotericin B and 5-flucytosine and warrants further evaluation. PMID- 10834996 TI - Diagnostic potential of puumala virus nucleocapsid protein expressed in Drosophila melanogaster cells. AB - Puumala virus (PUU) nucleocapsid protein (N) was expressed in insect cells by using the Drosophila Expression System (DES; Invitrogen BV, Groningen, The Netherlands). Stable transfectants were established by hygromycin B selection and showed continuous expression of the recombinant protein (DES-PUU-N) for at least 5 months. The antigenic property of DES-PUU-N was shown to be identical to that of native PUU N when examined with a panel of hantavirus-specific monoclonal antibodies. Enzyme-linked immunosorbent assays (ELISAs) for detection of human immunoglobulin M (IgM) and IgG antibodies were established by using DES-PUU-N as antigen and were compared to assays based on native N. The ELISAs were evaluated for patient diagnosis and seroepidemiological purposes with panels of sera collected from patients with hemorrhagic fever with renal syndrome (HFRS) and from healthy blood donors. Equally high sensitivities and specificities for detection of PUU-specific IgM in acute-phase HFRS patient sera were obtained by the ELISA based on DES-PUU-N and the assay based on the native antigen. For detection of PUU-specific IgG, the ELISA based on monoclonal antibody-captured DES-PUU-N antigen showed optimal sensitivity and specificity. PMID- 10834999 TI - Intervening transcribed spacer region 1 variability in Cyclospora cayetanensis. AB - Cyclospora cayetanensis is an apicomplexan protozoan parasite which has emerged as an important cause of epidemic and endemic diarrhea. Water-borne as well as food-borne outbreaks have occurred, including a large number of U.S. cases associated with raspberries imported from Guatemala. Molecular markers exist for tracing the epidemiology of many of the bacterial pathogens associated with water borne or food-borne diarrhea, such as serotyping and pulsed-field electrophoresis. However, there are currently no molecular markers available for C. cayetanensis. The intervening transcribed spacer (ITS) regions between the small- and large-subunit rRNA genes demonstrate much greater sequence variability than the small-subunit rRNA sequence itself and have been useful for the molecular typing of other organisms. Thus, ITS1 variability might allow the identification of different genotypes of C. cayetanensis. In order to determine the degree of ITS1 variability among C. cayetanensis isolates, the ITS1 sequences of C. cayetanensis isolates from a variety of sources, including raspberry associated cases, cases from Guatemala, and pooled and individual isolates from Peru, were obtained. The ITS1 sequences of all five raspberry-associated isolates were identical, consistent with their origin from a single source. In contrast, one of the two Guatemala isolates and two Peruvian isolates contained multiple ITS1 sequences. These multiple sequences could represent multiple clones from a single clinical source or, more likely, variability of the ITS1 region within the genome of a single clone. PMID- 10835000 TI - Diagnosis of invasive amebiasis by enzyme-linked immunosorbent assay of saliva to detect amebic lectin antigen and anti-lectin immunoglobulin G antibodies. AB - Saliva from subjects with amebic liver abscess (ALA), acute amebic colitis, asymptomatic infection with Entamoeba histolytica or Entamoeba dispar, and uninfected controls was tested by enzyme-linked immunosorbent assay (ELISA) for the presence of E. histolytica galactose-inhibitable lectin antigen and salivary immunoglobulin (IgG) antibodies to a recombinant cysteine-rich lectin-derived protein (LC3). Salivary lectin antigen was found in 65.8% of subjects with acute colitis, compared to 22.2% of those convalescent from ALA, 10.0% with asymptomatic E. histolytica infection, 9.8% with E. dispar infection, and 2.6% of controls (subjects from the United States and study patients with nonamebic diarrhea) (P < 0.001 for each compared to values for subjects with colitis). Salivary anti-LC3 IgG antibodies were found in 92% of ALA patients regardless of duration of illness and in 83.3% of colitis patients who were symptomatic for at least 7 days (P < 0.001 compared to other study groups). Serum anti-LC3 IgG antibodies were detected in 56.3% of subjects with acute colitis, 100% of subjects with ALA or prolonged colitis, 45% of subjects with asymptomatic E. histolytica infection, 32.3% of subjects with E. dispar infection, and 23.4% of diarrhea controls. In comparison to ELISA for serum anti-LC3 IgG antibodies, the salivary lectin antigen assay is a more sensitive and specific test for acute amebic colitis. Detection of salivary anti-LC3 IgG antibodies by ELISA is an effective means for the diagnosis of ALA and prolonged cases of amebic colitis. PMID- 10835001 TI - Multicenter evaluation of methods to quantitate human immunodeficiency virus type 1 RNA in seminal plasma. AB - We have evaluated two commercially available kits (AMPLICOR MONITOR [Roche] and NASBA HIV-1 QT or NucliSens HIV-1 QT [Organon Teknika]) and two noncommercial methods for the accurate quantitation of human immunodeficiency virus type 1 (HIV 1) RNA in seminal plasma. The same panels of coded specimens were tested on four separate occasions. Laboratories using the commercial assays employed silica beads to isolate HIV-1 RNA, which removed inhibitory factors sometimes found in seminal plasma. Sensitivities and specificities, respectively, for each assay were as follows: AMPLICOR MONITOR, 100 and 73%; NASBA HIV-1 QT, 84 and 100%; NucliSens HIV-1 QT, 99 and 98%; and noncommercial assays, 91 and 73%. When results from the laboratory that was inexperienced with the silica bead extraction method were excluded from the analysis, specificity for the Roche assay increased to 100%. The commercial assays demonstrated highly reproducible results, with intra-assay standard deviations (measured in log(10) RNA copies/milliliter of seminal plasma) ranging from 0.11 to 0.32; those of the noncommercial assays ranged from 0.12 to 0.75. Differences in mean estimated HIV 1 RNA concentrations were /=64 microgram/ml) isolates of Candida albicans from prospectively monitored HIV-infected children with OPEC were studied. FR isolates were recovered from children with severe OPEC refractory to fluconazole, and FS isolates were recovered from those with mucosal candidiasis responsive to fluconazole. Fluconazole at 2 mg/kg of body weight/day was administered to infected animals for 7 days. The concentrations of fluconazole in plasma were maintained above the MICs for FS isolates throughout the dosing interval. Fluconazole concentrations in the esophagus were greater than or equal to those in plasma. Rabbits infected with FS isolates and treated with fluconazole had significant reductions in oral mucosal quantitative cultures (P < 0.001) and tissue burden of C. albicans in tongue, soft palate, and esophagus (P < 0.001). In comparison, rabbits infected with FR isolates were unresponsive to fluconazole and had no reduction in oral mucosal quantitative cultures or tissue burden of C. albicans versus untreated controls. We conclude that there is a strong correlation between in vitro fluconazole susceptibility by NCCLS methods and in vivo response to fluconazole therapy of OPEC due to C. albicans. PMID- 10835006 TI - Isolation of Acanthamoeba-specific antibodies from a bacteriophage display library. AB - Acanthamoeba causes opportunistic eye infections in humans, which can lead to severe keratitis and may ultimately result in blindness. Current methods for identifying this organism rely on culture and microscopy. In this paper, we describe the isolation of antibody fragments that can be used for the unequivocal identification of Acanthamoeba. A bacteriophage antibody display library was used to isolate antibody fragments that bind specifically to Acanthamoeba. Individual clones were studied by enzyme-linked immunosorbent assay, flow cytometry, and immunofluorescence. Four antibody clones that specifically bind to Acanthamoeba spp. were identified. PMID- 10835007 TI - New chromogenic identification and detection of Staphylococcus aureus and methicillin-resistant S. aureus. AB - This paper describes a new chromogenic plate medium, CHROMagar Staph aureus (CHROMagar, Paris, France), for the identification of Staphylococcus aureus on the basis of colony pigmentation. The abilities of CHROMagar Staph aureus, thermostable nuclease (DNase), and mannitol salt agar (MSA) to identify S. aureus isolates (n = 114) and discriminate between S. aureus and coagulase-negative staphylococci (CoNS; n = 22) were compared. CHROMagar Staph aureus proved to be more sensitive and specific than DNase and MSA, allowing a reliable, simple, and rapid method for the identification of S. aureus isolates. All CoNS encountered in this study with the exception of S. chromogenes could be easily differentiated from S. aureus on this medium. The supplementation with 4 microgram of oxacillin or methicillin per ml allowed simple identification of methicillin resistance in hospital-acquired S. aureus strains which show multiple-drug resistance profiles. Community-acquired methicillin-resistant S. aureus strains showing non-multi-drug resistance profiles require further evaluation on this new chromogenic medium. Methicillin or oxacillin resistance of all S. aureus isolates was confirmed by the detection of penicillin-binding protein 2a, encoded by the mecA gene, using the latex slide agglutination MRSA-Screen test (PBP 2' Test, DR900M; Oxoid). PMID- 10835008 TI - Direct evidence by DNA fingerprinting that endoscopic cross-infection of Helicobacter pylori is a cause of postendoscopic acute gastritis. AB - The DNA fingerprinting of Helicobacter pylori strains in two cases of acute gastritis that occurred after endoscopy was examined. H. pylori was isolated from the stomachs of two patients with acute gastritis and from the stomachs of the patients in whom the same gastrofiberscope had previously been used. The genomic DNA digested with HaeIII was subjected to pulsed-field gel electrophoresis. The corresponding paired electrophoretic patterns were completely identical. These findings provide direct evidence that postendoscopic acute gastritis can be caused by cross-infection with H. pylori via endoscopy. PMID- 10835009 TI - Atypical lipid-dependent Malassezia species isolated from dogs with otitis externa. AB - During a survey of the occurrence of Malassezia species in the external ear canals of dogs with chronic otitis externa, lipid-dependent Malassezia species were isolated in three dogs. These species were identified as Malassezia furfur and M. obtusa but showed atypical assimilation patterns. To our knowledge, this is the first report of the isolation of lipid-dependent species of the genus Malassezia in association with canine otitis. PMID- 10835010 TI - Epidemiology of recurrences or reinfections of Clostridium difficile-associated diarrhea. AB - Approximately 15 to 35% of patients with a first episode of Clostridium difficile associated diarrhea relapse within 2 months. Between 1994 and 1997, strains from 93 hospitalized patients with C. difficile recurrences were fingerprinted by using both serotyping and PCR-ribotyping. The results showed that 48.4% of clinical recurrences were, in fact, reinfections with a different strain of C. difficile. Rates of clinical recurrences could therefore be reduced by implementing strict isolation precautions. PMID- 10835011 TI - Genetic homology among thirteen Encephalitozoon intestinalis isolates obtained from human immunodeficiency virus-infected patients with intestinal microsporidiosis. AB - The ribosomal DNA internal transcribed spacer sequences of 13 unrelated Encephalitozoon intestinalis isolates obtained from human immunodeficiency virus (HIV)-infected patients with intestinal microsporidiosis were analyzed by gene amplification and DNA sequencing. Among these isolates, we found only one genetic lineage which suggests that E. intestinalis may have a clonal distribution in HIV infected patients. PMID- 10835013 TI - Comparison of recoveries of mycobacterium tuberculosis using the automated BACTEC MGIT 960 system, the BACTEC 460 TB system, and Lowenstein-Jensen medium. AB - Using two different liquid media and one conventional solid medium, a total of 57 mycobacterial isolates (Mycobacterium tuberculosis, n = 55; nontuberculous mycobacteria, n = 2) were recovered from 377 clinical specimens. The rates of recovery of M. tuberculosis were 96. 4% with the BACTEC MGIT 960 liquid medium, 92.7% with BACTEC 12B liquid medium, and 81.8% with the Lowenstein-Jensen (LJ) medium. The mean time to detection of M. tuberculosis in smear-positive specimens was 12.6 days for BACTEC MGIT 960 medium, 13.8 days for BACTEC 12B medium, and 20.1 days for LJ medium, and in smear-negative specimens it was 15.8 days for BACTEC MGIT 960 medium, 17.7 days for BACTEC 12B medium, and 42.2 days for LJ medium. The rates of contamination were 3.7, 2.9, and 1.2% for the BACTEC MGIT 960, BACTEC 12B, and LJ media, respectively. In conclusion, the nonradiometric, fully automated 7-ml BACTEC MGIT 960 system can be considered a viable alternative to the semiautomated, radiometric BACTEC 460 TB system. PMID- 10835014 TI - Survival of the human granulocytic ehrlichiosis agent under refrigeration conditions. AB - The human granulocytic ehrlichiosis (HGE) agent in infected blood specimens remained viable during refrigeration at 4 degrees C for up to 18 days. These findings suggest that blood specimens submitted for culture may withstand transportation to a remote laboratory. HGE should be added to the list of infections potentially transmitted by blood transfusion. PMID- 10835012 TI - Molecular characterization of the vanD gene cluster and a novel insertion element in a vancomycin-resistant enterococcus isolated in Canada. AB - A single vanD-containing Enterococcus faecium strain (N97-330) was isolated in Canada. The vanD-containing region was cloned and sequenced. Although the proteins have more than 96% identity to a previously described vanD region in BM4339, the vanS(D) gene contains a frameshift mutation that leads to a predicted truncated protein. Furthermore, sequence analysis of the ddl gene revealed the presence of an IS982-like element (ISEfm1) which interrupted the D-Ala-D-Ala ligase. This suggested the constitutive expression of the vanD operon, which was confirmed. Pulsed-field gel electrophoresis fingerprinting demonstrated that BM4339 was not related to N97-330 (>15 band differences). Both strains contained multiple copies of the IS982-like element. PMID- 10835015 TI - Retrospective diagnosis of diphtheria by detection of the Corynebacterium diphtheriae tox gene in a formaldehyde-fixed throat swab using PCR and sequencing analysis. AB - The tox gene of Corynebacterium diphtheriae was detected in a formaldehyde-fixed throat swab taken from a 68-year-old man who was reported to have died of suffocation due to a pharyngeal tumor. DNA templates prepared from bacterial cells fixed with 10% formaldehyde were subjected to a PCR analysis with tox gene specific PCR primers. The resultant 112-nucleotide-long PCR product was sequenced using a dye terminator method, and an expected 57-nucleotide-long internal sequence of the tox gene was detected. This method is applicable for retrospective diagnosis in diphtheria cases in which only a formaldehyde-fixed clinical sample is available. PMID- 10835016 TI - Epidemiologic typing of Salmonella enterica serotype enteritidis in a Canada-wide outbreak of gastroenteritis due to contaminated cheese. AB - A major Canada-wide outbreak of gastroenteritis due to Salmonella enterica serotype Enteritidis phage type (PT) 8 occurred in 1998, and this was traced to contaminated cheese in a commercial lunch pack product. Phage typing and pulsed field gel electrophoresis linked the clinical and cheese isolates of serotype Enteritidis but failed to differentiate outbreak from nonoutbreak PT 8 strains. Further differentiation was made by biotyping based on melibiose fermentation. PMID- 10835017 TI - High prevalence of TT virus DNA in human saliva and semen. AB - Using the PCR assay, we found a high prevalence of TT virus (TTV) DNA in saliva and semen from patients who were seropositive for TTV. This finding suggests that the presence of TTV in body fluids other than serum may affect the routes of viral transmission. PMID- 10835018 TI - Limitations of vitek GPS-418 cards in exact detection of vancomycin-resistant enterococci with the vanB genotype. AB - The susceptibilities of 20 strains of vancomycin-resistant enterococci (VRE) with the vanB genotype obtained by using Vitek GPS-418 cards were compared with those obtained by the broth dilution method of the National Committee for Clinical Laboratory Standards (NCCLS) (approved standard M7-A4) and with those obtained by the agar screen method using bile esculin azide agar containing 6 microgram of vancomycin per ml. Although both the broth dilution and agar screen methods disclosed no discordance, Vitek GPS-418 cards yielded a very major error compared with the results obtained by the reference broth dilution method of the NCCLS. Vitek GPS-418 cards were therefore found to have considerable room for improvement for the accurate detection of vanB VRE strains. PMID- 10835019 TI - Identification of Arthrobacter oxydans, Arthrobacter luteolus sp. nov., and Arthrobacter albus sp. nov., isolated from human clinical specimens. AB - Five Arthrobacter isolates from clinical specimens were studied by phenotypic, chemotaxonomic, and genetic characterization. Two strains had characteristics consistent with those of Arthrobacter oxydans. One strain was related to A. citreus; however, DNA-DNA hybridization and phenotypic characteristics indicated that this strain belongs to a new species, for which the name Arthrobacter luteolus sp. nov. is proposed. Two strains were closely related to A. cumminsii by 16S rRNA gene sequencing, but DNA-DNA hybridization, peptidoglycan type, and some phenotypic features indicated that they should be assigned to a new species, for which the name Arthrobacter albus sp. nov. is proposed. The type strain of A. luteolus is CF25 (DSM 13067). The type strain of A. albus is CF43 (DSM 13068). PMID- 10835020 TI - Reproducibility problems with the Abbott laboratories LCx assay for Chlamydia trachomatis and Neisseria gonorrhoeae. AB - This study demonstrates that significant reproducibility problems can occur during routine use of the Abbott Laboratories LCx assay for Chlamydia trachomatis and Neisseria gonorrhoeae. These problems can go undetected by the quality control procedures outlined in the manufacturer's package insert. We outline here procedures for detecting and preventing contamination and reproducibility problems. PMID- 10835021 TI - Genetic similarity among one Aspergillus flavus strain isolated from a patient who underwent heart surgery and two environmental strains obtained from the operating room. AB - We report the simultaneous isolation of one Aspergillus flavus strain from the aortic prosthesis of a heart surgery patient and another two isolates recovered from a dual-reservoir cooler-heater used in the operating room where this patient was operated on. Genetic typing of these three isolates by randomly amplified polymorphic DNA (RAPD) revealed identical genotypes. Eight unrelated control strains of A. flavus had eight different genotypes. These results clearly indicated the nosocomial origin of the A. flavus strain isolated from the patient. We suggest that the RAPD technique is a rapid and reliable tool to ascertain the epidemiology of infections caused by A. flavus. PMID- 10835022 TI - Retrospective identification and characterization of Candida dubliniensis isolates among Candida albicans clinical laboratory isolates from human immunodeficiency virus (HIV)-infected and non-HIV-infected individuals. AB - Fungal opportunistic infections, and in particular those caused by the various Candida species, have gained considerable significance as a cause of morbidity and, often, mortality. The newly described species Candida dubliniensis phenotypically resembles Candida albicans so closely that it is easily misidentified as such. The present study was designed to determine the frequency at which this new species is not recognized in the clinical laboratory, to determine the patient populations with which C. dubliniensis is associated, to determine colonization versus infection frequency, and to assess fluconazole resistance. Over a 2-year period, 1,251 isolates that were initially identified as C. albicans by a hospital clinical laboratory were reevaluated for C. dubliniensis by inability to grow at 45 degrees C, colony color on CHROMagar Candida medium, coaggregation assay with Fusobacterium nucleatum, and sugar assimilation profiles (API 20C AUX yeast identification system). A total of 15 (1.2%) isolates from 12 patients were identified as C. dubliniensis. Ten of the patients were found to be immunocompromised (these included patients with human immunodeficiency virus infection or AIDS, cancer patients receiving chemotherapy, and patients awaiting transplantation). Thirteen isolates were highly susceptible to fluconazole (MIC, <0.5 microgram/ml). Three isolates from one patient, genotypically confirmed as the same strain, showed variable susceptibility to fluconazole. The first isolate was susceptible, whereas the other two isolates were dose-dependent susceptible (MIC, 16.0 microgram/ml). These data confirm the close association of C. dubliniensis with immunocompromised states and that increased fluconazole MICs may develop in vivo. This study emphasizes the importance of screening germ-tube-positive yeasts for the inability to grow at 45 degrees C followed by confirmatory tests in order to properly identify this species. PMID- 10835023 TI - Limitation of the AccuProbe Coccidioides immitis culture identification test: false-negative results with formaldehyde-killed cultures. AB - The AccuProbe Coccidioides immitis culture identification test (CI test) yielded false-negative results with formaldehyde-killed C. immitis submitted to a reference laboratory. Further evaluation with pure or mixed cultures or stored, heat-killed cultures revealed the CI test to be highly sensitive and specific for C. immitis except when the cultures were pretreated with formaldehyde. PMID- 10835024 TI - Rapid identification of methicillin-resistant Staphylococcus aureus and simultaneous species confirmation using real-time fluorescence PCR. AB - A duplex LightCycler PCR assay targeting the mecA gene and a Staphylococcus aureus-specific marker was used to test 165 S. aureus strains and 80 strains of other bacterial species. Within an assay time of 60 min plus 10 min for sample preparation, S. aureus as well as the presence or absence of the mecA gene was correctly identified. PMID- 10835025 TI - Phaeoisaria clematidis as a cause of keratomycosis. AB - We report the first case of human infection by Phaeoisaria clematidis. This fungus caused a corneal ulcer in a Brazilian man who had previously suffered an eye injury. Diagnosis was established by positive direct examination and repeated cultures. The isolate was clearly resistant in vitro to the six antifungal agents tested. PMID- 10835026 TI - Culture of Helicobacter pylori from a gastric string may be an alternative to endoscopic biopsy. AB - Helicobacter pylori was isolated from a swallowed string from 32 of 33 adult subjects (97%) with selective culture media. With this method, antibiotic susceptibility testing and molecular epidemiology studies of H. pylori can be carried out without the need for the collection of specimens by endoscopic biopsy. PMID- 10835027 TI - Relationship of genetic type of Shiga toxin to manifestation of bloody diarrhea due to enterohemorrhagic Escherichia coli serogroup O157 isolates in Osaka City, Japan. AB - One hundred sixty-nine strains of enterohemorrhagic Escherichia coli serogroup O157 were examined for the correlation between the genotype of their Shiga toxin genes (stx) and manifestation of bloody diarrhea (BD). It was shown that the strains carrying only stx2vha were probably less virulent and caused BD less frequently. PMID- 10835029 TI - Comparison of quantitations of viral load in varicella and zoster. AB - The pathogenesis of varicella and zoster and the effects of antiviral treatment were investigated using real-time PCR for varicella-zoster virus (VZV) DNA in skin lesions and peripheral blood. A higher occurrence of viremic VZV DNA was observed in varicella than in zoster. Acyclovir treatment resulted in marked suppression of viremia in varicella. PMID- 10835028 TI - Molecular characterization of porcine rotaviruses from the southern region of Brazil: characterization of an atypical genotype G[9] strain. AB - The G (VP7) and P (VP4) serotype distribution of Brazilian porcine rotaviruses was determined using reverse transcription-PCR genotyping methods. Common porcine G types G3, G4, and G5 were detected in combination with P types [6] and [7]. The detection of nonporcine G types and unusual G-P combinations and the characterization of an atypical virus indicated that interspecies transmission may contribute to the genetic diversity of porcine rotaviruses. PMID- 10835030 TI - Recurrent bacteremic peritonitis caused by Enterococcus cecorum in a patient with liver cirrhosis. AB - Enterococcus cecorum (formerly Streptococcus cecorum), originally isolated from poultry intestines, has rarely been encountered in human diseases. A 60-year-old man with liver cirrhosis and hepatocellular carcinoma developed peritonitis on the seventh day of his hospitalization. Cultures of one blood sample and one ascites fluid sample obtained on that day both grew E. cecorum. The patient received intravenous cefoxitin therapy and initially responded well. Unfortunately, another episode of peritonitis associated with septic shock developed 24 days after the start of treatment, and culture of one blood specimen yielded the same organism. The isolates were identified by the conventional biochemical tests, the API Rapid ID 32 Strep system, and the API ZYM system (both systems from bioMerieux, Marcy L'Etoile, France) and were further confirmed by cellular fatty acid chromatography and 16S rRNA gene partial sequencing. The identical biotype, antibiotype, and random amplified polymorphic DNA pattern of the three isolates documented the long-term persistence of this organism in the patient. To the best of our knowledge, this is the first clinical description of recurrent bacteremic peritonitis caused by E. cecorum. PMID- 10835031 TI - Genetic characterization of an H1N2 influenza virus isolated from a pig in Indiana. AB - An H1N2 influenza virus was isolated from a pig during an outbreak of respiratory disease and abortion on an Indiana farm in November 1999. Results of phylogenetic analyses indicate that this virus is a reassortant between a recent classical H1 swine virus and the reassortant H3N2 viruses that have emerged among American pigs since 1998. PMID- 10835032 TI - A slow NMDA channel: in search of a role. AB - Recombinant NMDA receptors comprising NR1/NR2D subunits have unique properties when compared with other subtypes. These include low channel conductance with a characteristic pattern of transition between subconductance states and reduced sensitivity to Mg2+ blockade. Native NMDA receptors matching the low conductance and Mg2+ sensitivity of NR1/NR2D recombinant channels have been found in a few neuronal populations, in particular in developing Purkinje neurons (Momiyama et al. 1996). Ultra-rapid agonist application by means of a piezoelectric translator to excised outside-out membrane patches allows concentration jumps in the milliseconds range similar to those occurring during vesicular synaptic release. The high affinity of glutamate for NR1/NR2D receptors produces an extremely slow channel deactivation in response to brief glutamate applications (Monyer et al. 1994). This property has now been identified for the first time by Misra et al. (this issue of The Journal of Physiology, Misra et al. 2000b) for native NMDA channels in mammalian neurons. PMID- 10835034 TI - Slow deactivation kinetics of NMDA receptors containing NR1 and NR2D subunits in rat cerebellar Purkinje cells. AB - We have examined the deactivation kinetics of native N-methyl-D-aspartate receptors (NMDARs) containing NR1 and NR2D subunits by patch-clamp recording from Purkinje cells in cerebellar slices from young rats. NMDAR-mediated whole-cell currents were elicited in response to bath application of 20 microM NMDA and 50 microM glycine. The NMDAR-mediated currents were small, with an average whole cell conductance of approximately 750 pS. Following the rapid application of brief pulses (1-10 ms) of 1 mM glutamate to outside-out membrane patches, we observed a low-conductance type of single-channel activity which lasted up to 30 s after the removal of agonist. Analysis of individual channel openings revealed asymmetry of transitions between the main- and subconductance states - a characteristic of NR1/NR2D-containing NMDARs. The averaged macroscopic current exhibited a decay time course which was well described by a single exponential function with a time constant of approximately 3 s. We conclude that native NR1/NR2D-containing NMDARs, like their recombinant counterparts, display very slow deactivation kinetics. This feature should provide a means for identification of these receptors at synapses, and indicates that they do not contribute to the synaptic NMDAR currents so far described. PMID- 10835035 TI - ATP-sensitive potassium channels in capillaries isolated from guinea-pig heart. AB - The full-length cDNAs of two different alpha-subunits (Kir6.1 and Kir6.2) and partial cDNAs of three different beta-subunits (SUR1, SUR2A and SUR2B) of ATP sensitive potassium (KATP) channels of the guinea-pig (gp) were obtained by screening a cDNA library from the ventricle of guinea-pig heart. Cell-specific reverse-transcriptase PCR with gene-specific intron-spanning primers showed that gpKir6.1, gpKir6.2 and gpSUR2B were expressed in a purified fraction of capillary endothelial cells. In cardiomyocytes, gpKir6.1, gpKir6.2, gpSUR1 and gpSUR2A were detected. Patch-clamp measurements were carried out in isolated capillary fragments consisting of 3-15 endothelial cells. The membrane capacitance measured in the whole-cell mode was 19.9 +/- 1.0 pF and was independent of the length of the capillary fragment, which suggests that the endothelial cells were not electrically coupled under our experimental conditions. The perforated-patch technique was used to measure the steady-state current-voltage relation of capillary endothelial cells. Application of K+ channel openers (rilmakalim or diazoxide) or metabolic inhibition (250 microM 2,4-dinitrophenol plus 10 mM deoxyglucose) induced a current that reversed near the calculated K+ equilibrium potential. Rilmakalim (1 microM), diazoxide (300 microM) and metabolic inhibition increased the slope conductance measured at -55 mV by a factor of 9.0 (+/-1.8), 2.5 (+/-0.2) and 3.9 (+/-1.7), respectively. The effects were reversed by glibenclamide (1 microM). Our results suggest that capillary endothelial cells from guinea-pig heart express KATP channels composed of SUR2B and Kir6.1 and/or Kir6.2 subunits. The hyperpolarization elicited by the opening of KATP channels may lead to an increase in free cytosolic Ca2+, and thus modulate the synthesis of NO and the permeability of the capillary wall. PMID- 10835033 TI - Molecular basis of functional voltage-gated K+ channel diversity in the mammalian myocardium. AB - In the mammalian heart, Ca2+-independent, depolarization-activated potassium (K+) currents contribute importantly to shaping the waveforms of action potentials, and several distinct types of voltage-gated K+ currents that subserve this role have been characterized. In most cardiac cells, transient outward currents, Ito,f and/or Ito,s, and several components of delayed reactivation, including IKr, IKs, IKur and IK,slow, are expressed. Nevertheless, there are species, as well as cell type and regional, differences in the expression patterns of these currents, and these differences are manifested as variations in action potential waveforms. A large number of voltage-gated K+ channel pore-forming (alpha) and accessory (beta, minK, MiRP) subunits have been cloned from or shown to be expressed in heart, and a variety of experimental approaches are being exploited in vitro and in vivo to define the relationship(s) between these subunits and functional voltage-gated cardiac K+ channels. Considerable progress has been made in defining these relationships recently, and it is now clear that distinct molecular entities underlie the various electrophysiologically distinct repolarizing K+ currents (i.e. Ito,f, Ito,s, IKr, IKs, IKur, IK,slow, etc.) in myocyardial cells. PMID- 10835036 TI - Desensitisation of protease-activated receptor-1 (PAR-1) in rat astrocytes: evidence for a novel mechanism for terminating Ca2+ signalling evoked by the tethered ligand. AB - Protease-activated receptor-1 (PAR-1), a G-protein-coupled receptor, is activated when thrombin cleaves its N-terminal exodomain, thereby regulating morphology, growth and survival of neurones and astrocytes. We have investigated the mechanism of PAR-1 desensitisation and resensitisation after proteolytic or non proteolytic stimulation with thrombin or thrombin receptor agonist peptide (TRag), respectively. In rat primary astrocytes, short-term stimulation with thrombin resulted in a single [Ca2+]i transient and a dose-dependent de- and resensitisation, as assessed by single-cell Ca2+ imaging of fura-2-loaded astrocytes. An initial proteolytic activation of astrocyte PAR-1 by exposure to thrombin strongly decreased the response elicited by subsequent application of a second dose of thrombin or of TRag. In contrast, after an initial non-proteolytic activation of astrocyte PAR-1 by TRag, the subsequent response to thrombin, but not to an additional application of TRag, was strongly attenuated, and the time course for desensitisation was slower. Based on this finding we hypothesised that after PAR-1 activation, the 'tethered ligand' is proteolytically destroyed. As a consequence, the receptor becomes unresponsive to a subsequent thrombin stimulus but is still capable of responding to TRag. This hypothesis was supported by applying thermolysin, which is known to cleave PAR-1 within its tethered-ligand domain, and was confirmed by incubation with soybean trypsin inhibitor. PAR-1 resensitisation occurs via new PAR-1 synthesis since resensitisation was inhibited by cycloheximide and brefeldin A. From these results, we derive a novel model wherein activation of PAR-1, in addition to initiating signal transduction, activates a protease mechanism that cleaves the N-terminus of the receptor, thus terminating the signal and probably inducing receptor internalisation. PMID- 10835037 TI - Presynaptic dopamine D2 and muscarine M3 receptors inhibit excitatory and inhibitory transmission to rat subthalamic neurones in vitro. AB - Whole-cell patch-clamp recordings were made from subthalamic nucleus (STN) neurones in brain slices from rats. Stimulation with bipolar electrodes evoked synaptic currents mediated by glutamate (EPSCs) and GABAA (IPSCs) receptors. Dopamine reversibly reduced the amplitude of GABAA IPSCs by up to 48 % with an IC50 value of 3.4 +/- 0.8 microM. The dopamine D2 receptor agonist quinpirole, but not the D1 receptor agonist SKF 82958, also inhibited GABAA IPSCs. This effect was completely reversed by the D2 receptor antagonist sulpiride but not by SCH 23390, a D1 antagonist. Muscarine reversibly reduced the amplitude of GABAA IPSCs by up to 70 % with an IC50 value of 0.6 +/- 0.1 microM. Inhibition of IPSCs by muscarine was completely blocked by scopolamine (10 microM), a muscarinic receptor antagonist. The M3 muscarinic receptor antagonist 4-DAMP effectively reversed muscarine-induced inhibition of IPSCs with an IC50 of 0.11 +/- 0.03 microM. Although the M1 receptor antagonist pirenzepine also reversed the inhibition of IPSCs by muscarine, this effect was only observed at relatively high concentrations (IC50 = 21.7 +/- 9.4 microM). Dopamine and muscarine both increased the paired-pulse ratio of GABAA IPSCs. Neither agent produced sustained changes in postsynaptic holding current. Glutamate EPSCs were also inhibited reversibly by dopamine (by up to 29%; IC50 = 16 +/- 3 microM) and muscarine (by up to 41%; IC50 = 1.0 +/- 0.4 microM). However, both agents were more potent and efficacious for reducing GABA IPSCs compared with glutamate EPSCs. These results suggest that the most significant effect of dopamine and muscarine in the STN is to reduce inhibitory synaptic input by acting at presynaptic dopamine D2 and muscarinic M3 receptors, respectively. PMID- 10835038 TI - Regulation of Na+,K+-ATPase by persistent sodium accumulation in adult rat thalamic neurones. AB - The present study investigated the regulatory mechanism of the Na+, K+-ATPase and the level of internal Na+ and Ca2+ in response to persistent Na+ influx in acutely dissociated rat thalamic neurones. Whole-cell patch-clamp recordings and Na+ imaging revealed a stable [Na+]i and low background pump activity. Exposure to veratridine (50 microM) for 1 h resulted in a progressive rise in [Na+]i (DeltaFNa = 64 +/-22%) and [Ca2+]i (DeltaFCa = 44 +/- 14%) over 3 h. Increases in [Na+]i and [Ca2+]i were also observed during neuronal exposure to the Na+ ionophore monensin (50 microM). Subcellular confocal immunofluorescence quantification of alpha3 catalytic Na+-K+ pump subunits showed that a veratridine induced rise in [Na+]i was accompanied by a significant increase in pump density in both membrane and cytoplasmic compartments, by 39 and 54%, respectively. Similar results were also obtained in experiments when neurones were treated with monensin. A fluorescent 9-anthroylouabain binding assay detected a 60 and 110% increase in phosphorylated (active) pumps after veratridine and monensin exposure, respectively. During the entire experiment, application of ouabain or veratridine alone induced little cell swelling and death, but pump inhibition in cells pre-loaded with Na+ led to rapid cell swelling and necrosis. The above results indicate that a persistent influx of Na+ may trigger rapid enhancement of pump synthesis, membrane redistribution and functional activity. However, these compensatory mechanisms failed to prevent persistent Na+ accumulation. PMID- 10835039 TI - Pacemaking in interstitial cells of Cajal depends upon calcium handling by endoplasmic reticulum and mitochondria. AB - Pacemaker cells, known as interstitial cells of Cajal (ICC), generate electrical rhythmicity in the gastrointestinal tract. Pacemaker currents in ICC result from the activation of a voltage-independent, non-selective cation conductance, but the timing mechanism responsible for periodic activation of the pacemaker current is unknown. Previous studies suggest that pacemaking in ICC is dependent upon metabolic activity 1y1yand1 Ca2+ release from intracellular stores. We tested the hypothesis that mitochondrial Ca2+ handling may underlie the dependence of gastrointestinal pacemaking on oxidative metabolism. Pacemaker currents occurred spontaneously in cultured ICC and were associated with mitochondrial Ca2+ transients. Inhibition of the electrochemical gradient across the inner mitochondrial membrane blocked Ca2+ uptake and pacemaker currents in cultured ICC and blocked slow wave activity in intact gastrointestinal muscles from mouse, dog and guinea-pig. Pacemaker currents and rhythmic mitochondrial Ca2+ uptake in ICC were also blocked by inhibitors of IP3-dependent release of Ca2+ from the endoplasmic reticulum and by inhibitors of endoplasmic reticulum Ca2+ reuptake. Our data suggest that integrated Ca2+ handling by endoplasmic reticulum and mitochondria is a prerequisite of electrical pacemaking in the gastrointestinal tract. PMID- 10835040 TI - Nitric oxide and thiol reagent modulation of Ca2+-activated K+ (BKCa) channels in myocytes of the guinea-pig taenia caeci. AB - The modulation of large conductance Ca2+-activated K+ (BKCa) channels by the nitric oxide (NO) donors S-nitroso-L-cysteine (NOCys) and sodium nitroprusside (SNP) and agents which oxidize or reduce reactive thiol groups were compared in excised inside-out membrane patches of the guinea-pig taenia caeci. When the cytosolic side of excised patches was bathed in a physiological salt solution (PSS) containing 130 mM K+ and 15 nM Ca2+, few BKCa channel openings were recorded at potentials negative to 0 mV. However, the current amplitude and open probability (NPo) of these BKCa channels increased with patch depolarization. A plot of ln(NPo) against the membrane potential (V) fitted with a straight line revealed a voltage at half-maximal activation (V0.5) of 9.4 mV and a slope (K) indicating an e-fold increase in NPo with 12.9 mV depolarization. As the cytosolic Ca2+ was raised to 150 nM, V0.5 shifted 11.5 mV in the negative direction, with little change in K (13.1 mV). NOCys (10 microM) and SNP (100 microM) transiently increased NPo 16- and 3. 7-fold, respectively, after a delay of 2-5 min. This increase in NPo was associated with an increase in the number of BKCa channel openings evoked at positive potentials by ramped depolarizations (between -60 and +60 mV). Moreover, this NOCys-induced increase in NPo was still evident in the presence of 1H-[1,2,4]oxadiazolo[4, 3-a]quinoxalin-1-one (ODQ; 10 microM), the specific blocker of soluble guanylyl cyclase. The sulfhydryl reducing agents dithiothreitol (DTT; 10 and 100 microM) and reduced glutathione (GSH; 1 mM) also significantly increased NPo (at 0 mV) 7- to 9-fold, as well as increasing the number of BKCa channel openings evoked during ramped depolarizations. Sulfhydryl oxidizing agents thimerosal (10 microM) and 4,4' dithiodipyridine (4,4DTDP; 10 microM) and the thiol-specific alkylating agent N ethylmaleimide (NEM; 1 mM) significantly decreased NPo (at 0 mV) to 40-50% of control values after 5-10 min. Ramped depolarizations to +100 mV evoked relatively few BKCa channel openings. The effects of thimerosal on NPo were readily reversed by DTT, while the effects of NOCys were prevented by NEM. It was concluded that both redox modulation and nitrothiosylation of cysteine groups on the cytosolic surface of the alpha subunit of the BKCa channel protein can alter channel gating. PMID- 10835041 TI - The alpha1B Ca2+ channel amino terminus contributes determinants for beta subunit mediated voltage-dependent inactivation properties. AB - Co-expression of auxiliary beta subunits with the alpha1B Ca2+ channel subunit in COS-7 cells resulted in an increase in current density and a hyperpolarising shift in the mid-point of activation. Amongst the beta subunits, beta2a in particular, but also beta4 and beta1b caused a significant retardation of the voltage-dependent inactivation compared to currents with alpha1B alone, whilst no significant changes in inactivation properties were seen for the beta3 subunit in this system. Prevention of beta2a palmitoylation, by introducing cysteine to serine mutations (beta2a(C3,4S)), greatly reduced the ability of beta2a to retard voltage-dependent inactivation. Deletion of the proximal half of the alpha1B cytoplasmic amino terminus (alpha1BDelta1-55) differentially affected beta subunit-mediated voltage-dependent inactivation properties. These effects were prominent with the beta2a subunit and, to a lesser extent, with beta1b. For beta2a, the major effects of this deletion were a partial reversal of beta2a mediated retardation of inactivation and the introduction of a fast component of inactivation, not seen with full-length alpha1B. Deletion of the amino terminus had no other major effects on the measured biophysical properties of alpha1B when co-expressed with beta subunits. Transfer of the whole alpha1B amino terminus into alpha1C (alpha1bCCCC) conferred a similar retardation of inactivation on alpha1C when co-expressed with beta2a to that seen in parental alpha1B. Individual (alpha1B(Q47A) and alpha1B(R52A)) and double (alpha1B(R52,54A)) point mutations within the amino terminus of alpha1B also opposed the beta2a-mediated retardation of alpha1B inactivation kinetics. These results indicate that the alpha1B amino terminus contains determinants for beta subunit-mediated voltage dependent inactivation properties. Furthermore, effects were beta subunit selective. As deletion of the alpha1B amino terminus only partially opposed beta subunit-mediated changes in inactivation properties, the amino terminus is likely to contribute to a complex site necessary for complete beta subunit function. PMID- 10835042 TI - Effects of arachidonic acid on unitary calcium currents in rat sympathetic neurons. AB - We have characterized the actions of arachidonic acid (AA) on whole cell and unitary calcium (Ca2+) currents in rat neonatal superior cervical ganglion (SCG) neurons using barium (Ba2+) as the charge carrier. Whole cell currents were elicited by stepping the membrane potential from -90 mV to +10 mV. Arachidonic acid (5 microM) was introduced into the bath in the continued presence of 1 microM (+)-202-791, an L-type Ca2+ channel agonist. Under these conditions, the peak current, comprised mainly of N-type current, and a slow, (+)-202-791-induced component of the tail current were inhibited by 67 +/- 6 and 60 +/- 10 %, respectively, indicating that AA inhibits both N- and L-type currents. At a test potential of +30 mV, AA (5 microM) decreased unitary L- and N-type Ca2+ channel open probability (Po) in cell-attached patches that contained a single channel. For both channels, the underlying causes of the decrease in Po were similar. Arachidonic acid caused an increase in the percentage of null sweeps and in the number of null sweeps that clustered together. In sweeps with activity, the average number of openings per sweep decreased, while first latency and mean closed time increased. Arachidonic acid had no significant effect on unitary current amplitude or mean open time. Our findings are the first description of the inhibition of unitary L- and N-type Ca2+ channel activity by AA and are consistent with both channels spending more time in their null mode and with increased dwell time in one or more closed states. PMID- 10835043 TI - Differences in Ca2+ buffering properties between excitatory and inhibitory hippocampal neurons from the rat. AB - Endogenous calcium binding ratios (kappaS) in dendrites of cultured hippocampal neurons were estimated according to the single compartment model for transients in intracellular Ca2+ concentration ([Ca2+]). In addition, the electrophysiological characteristics of neurons were classified by their autaptic currents and intrinsic firing patterns. These data were analysed in order to determine whether a correlation between Ca2+ buffers and electrophysiological type exists. Ca2+ binding ratios of endogenous buffers were estimated by eliciting [Ca2+] transients with short depolarizations, while cells were loaded with fura-2. Two types of estimates could be obtained: one termed kappaS(tau), based on analysing time constants (tau) of [Ca2+] transients, and another termed kappaS(dCa), derived from an analysis of initial amplitudes of [Ca2+] transients. Values for kappaS(tau) and kappaS(dCa) were estimated as 57 +/- 10 (mean +/- s.d., n = 10) and 60 +/- 14 (n = 10), respectively, in excitatory neurons, and 130 +/- 50 (n = 11) and 150 +/- 70 (n = 11), respectively, in inhibitory neurons. The kappaS values of excitatory and inhibitory cells were significantly different from each other, regardless of the measurement method (Student's t test, P < 0.01). However, there was no significant difference in kappaS between the groups classified according to firing patterns. Although kappaS(tau) values were well matched to those of kappaS(dCa) in most excitatory cells, the two values did not agree in three out of the fourteen inhibitory cells investigated. In these cells, the first few [Ca2+] transients after obtaining the whole cell configuration displayed a double exponential decay, suggesting that buffers with slow binding kinetics, such as parvalbumin, are involved. This hypothesis is further explored in an accompanying paper. PMID- 10835044 TI - Kinetics of Ca2+ binding to parvalbumin in bovine chromaffin cells: implications for [Ca2+] transients of neuronal dendrites. AB - kappaS1. The effect of parvalbumin (PV) on [Ca2+] transients was investigated by perfusing adrenal chromaffin cells with fura-2 and fluorescein isothiocyanate (FITC)-labelled PV. As PV diffused into cells, the decay of [Ca2+] transients was transformed from monophasic into biphasic. The proportion of the initial fast decay phase increased in parallel with the fluorescence intensity of FITC, indicating that PV is responsible for the initial fast decay phase. The relationship between the fast decay phase and the [Ca2+] level was investigated using depolarizing trains of stimuli. Within a train the relative amplitude of the fast decay phase was inversely dependent on the [Ca2+] level preceding a given stimulus. Based on these observations, we estimated the Ca2+ binding ratio of PV (kappaP), the apparent dissociation constant of PV for Ca2+ (Kdc, app), and the unbinding rate constant of Ca2+ from PV (kc-) in the cytosol of chromaffin cells. Assuming free [Mg2+] to be 0.14 mM, we obtained values of 51.4 +/- 2.0 nM (n = 3) and 0.95 +/- 0.026 s-1 (n = 3), for Kdc,app and kc-, respectively. With the parameters obtained in the perfusion study, we simulated [Ca2+] transients, using two different Ca2+ extrusion rates (gamma) - 20 and 300 s-1 - which represent typical values for chromaffin cells and neuronal dendrites, respectively. The simulation indicated that Ca2+ is pumped out before it is equilibrated with PV, when gamma is comparable to the equilibration rates between PV and Ca2+, resulting in the fast decay phase of a biexponential [Ca2+] transient. From these results we conclude that Ca2+ buffers with slow kinetics, such as PV, may cause biexponential decays in [Ca2+] transients, thereby complicating the analysis of endogenous Ca2+ binding ratios (kappaS) based on time constants. Nevertheless, estimates of kappaS based on Ca2+ increments provide reasonable estimates for Ca2+ binding ratios before equilibration with PV. PMID- 10835045 TI - Calcium dynamics and buffering in oculomotor neurones from mouse that are particularly resistant during amyotrophic lateral sclerosis (ALS)-related motoneurone disease. AB - Motoneurones are particularly vulnerable both in human forms of amyotrophic lateral sclerosis (ALS) and corresponding animal models of the disease. While most motoneurone populations are selectively impaired, oculomotor neurones are essentially resistant to ALS-related damage. Motoneurone vulnerability has been closely linked to disruptions of calcium signalling. To investigate underlying events, we performed a quantitative analysis of calcium homeostasis in oculomotor neurones from mice by simultaneous patch-clamp recordings in sliced tissue and microfluorometric-calcium measurements. Somatic calcium dynamics were investigated by using a computer-controlled microfluorometric system. In oculomotor neurones, basal calcium concentrations were around 80 nM and depolarisation-induced calcium responses were observed for membrane voltages positive to -40 u1u1u approximately mV1 approximately . Endogenous calcium homeostasis was quantified by using the 'added buffer' approach. The recovery phase of depolarisation-induced calcium transients was well approximated by a mono-exponential function with a decay time constant that showed a linear dependence on dye concentration. The extrapolated time constant in the absence of indicator dye was 1.7 +/- 0.2 s (n = 11 cells, 21C). Endogenous calcium binding ratios (kappa(s)) were found to be 264 +/- 25 (n = 11 cells), indicating that 99.6 % of cytosolic calcium ions were taken up by endogenous buffers. Recovery of calcium transients was characterised by an 'effective' extrusion rate gamma = 156 +/- 20 s-1 (n = 11 cells, 21 C). Endogenous calcium binding ratios in oculomotor neurones were 5- to 6-fold larger compared with those of more vulnerable motoneurones in the nucleus hypoglossus and spinal cord. In a first order approximation, they reduced the volume of local calcium elevations around open calcium channels, lowered peak amplitudes of global calcium transients for a given influx and prolonged calcium recovery times for a given set of uptake and extrusion mechanisms. With respect to motoneurone degeneration, our measurements suggest that the exceptional stability of oculomotor neurones partially results from a specialised calcium homeostasis based on high buffering capacities. Furthermore, they indicate that cellular adaptations that account for rapid calcium signalling in hypoglossal and spinal motoneurones enhance their vulnerability during ALS-related motoneurone disease. PMID- 10835046 TI - GABA mediates presynaptic inhibition at glycinergic synapses in a rat auditory brainstem nucleus. AB - Many inhibitory nerve terminals in the mammalian anteroventral cochlear nucleus (AVCN) contain both glycine and GABA, but the reason for the co-localization of these two inhibitory neurotransmitters in the AVCN is unknown. We have investigated the roles of glycine and GABA at synapses on bushy cells in the rat AVCN, using receptor immunohistochemistry and electrophysiology. Our immunohistochemical results show prominent punctate labelling of postsynaptic clusters of glycine receptors and of the receptor clustering protein gephyrin over the surface of bushy cells. In contrast, weak diffuse membrane immunolabelling of GABAA receptors was observed. Whole-cell recordings from bushy cells in AVCN slices demonstrated that evoked inhibitory postsynaptic currents (IPSCs) were predominantly (81 %) glycinergic, based on the decrease in amplitude of the IPSCs in bicuculline (10 microM). This observation was supported by the effect of strychnine (1 microM), which was to decrease the evoked IPSC (to 10 % of control IPSC amplitude) and to produce a greater than 90 % block of spontaneous miniature IPSCs. These results suggest a minor role for postsynaptic GABAA receptors in bushy cells, despite a high proportion of GABA-containing terminals on these cells. Therefore, a role for metabotropic GABAB receptors was investigated. Activation of GABAB receptors with baclofen revealed a significant attenuation of evoked glycinergic IPSCs. The effect of baclofen was presynaptic, as indicated by a lack of change in the mean amplitude of spontaneous IPSCs. Significantly, the decrease in the amplitude of evoked glycinergic IPSCs observed following repetitive nerve stimulation was reduced in the presence of the GABAB antagonist, CGP 35348. This indicates that synaptically released GABA can activate presynaptic GABAB receptors to reduce transmitter release at glycinergic synapses. Our results suggest specific pre- versus postsynaptic physiological roles for GABA and glycine in the AVCN. PMID- 10835047 TI - Dihydropyridine-induced Ca2+ release from ryanodine-sensitive Ca2+ pools in human skeletal muscle cells. AB - Dihydropyridines (DHPs) are widely used antihypertensive drugs and inhibit excitation-contraction (E-C) coupling in vascular smooth muscle and in myocardial cells by antagonizing L-type Ca2+ channels (DHP receptors). However, contradictory reports exist about the interaction of the DHP with the skeletal muscle isoform of the DHP receptor and E-C coupling in skeletal muscle cells. Using the intracellular fluorescent Ca2+ indicator fura-2, an increase in [Ca2+]i was observed after extracellular application of nifedipine to cultured human skeletal muscle cells. The rise in [Ca2+]i was dose dependent with a calculated EC50 of 614 +/- 96 nM nifedipine and a maximum increment in [Ca2+]i of 80 +/- 3.2 nM. Similar values were obtained with nitrendipine. This effect of DHPs was restricted to differentiated skeletal muscle cells and was not seen in non differentiated cells or in PC12 cells. In spite of the observed increase in [Ca2+]i, whole-cell patch clamp experiments revealed that 10 microM nifedipine abolished inward Ba2+ currents through L-type Ca2+ channels completely. Similar to nifedipine, (+/-)Bay K 8644, an agonist of the L-type Ca2+ channel, also increased [Ca2+]i. This effect could not be enhanced by further addition of nifedipine, suggesting that both DHPs act via a common signalling pathway. Based on the specific mechanism of the skeletal muscle E-C coupling, we propose the stabilization of a conformational state of the DHP receptor by DHPs, which is sufficient to activate the ryanodine receptor. PMID- 10835048 TI - G protein-mediated FMRFamidergic modulation of calcium influx in dissociated heart muscle cells from squid, Loligo forbesii. AB - The actions of the neuropeptide FMRFamide (Phe-Met-Arg-Phe-NH2) on the L-type (ICa,L) and T-type (ICa,T) calcium currents were investigated in muscle cells dissociated from the heart of squid, Loligo forbseii. The heart muscle cells could be divided into type I and type II cells, on the basis of morphological differences in the dissociated myocytes. FMRFamide induced a substantial block of the L-type calcium current seen in type I cells; this inhibition was rapid, reversible and dose dependent (IC50 = 0.1 microM). FMRFamide induced an increase in the amplitude of the L-type calcium current in the type II heart muscle cells, but had no effect on the T-type calcium current in either type of dissociated heart muscle cell, even at concentrations much higher than those found to affect the L-type calcium current. Internal dialysis of isolated type I heart muscle cells with guanosine 5'-O-(3-thiotriphosphate (GTPgammaS, 100 microM), a non hydrolysable GTP analogue, mimicked the FMRFamide inhibition of the Ca2+ current and occluded any further FMRFamide-induced inhibition. Internal dialysis of these cells with guanosine 5'-O-(2-thiodiphosphate) (GDPbetaS, 100 microM) reduced the FMRFamide-induced inhibition of the peak Ca2+ current. The inhibitory effects of FMRFamide were abolished by pre-incubation of the cells with pertussis toxin (200 ng ml-1). The activation kinetics of ICa,L were not affected by FMRFamide application, nor by internal perfusion with GTPgammaS, and the FMRFamide-induced reduction in ICa,L was not relieved by large depolarising prepulses. These data indicate that FMRFamide can modulate ICa,L, but not ICa,T, in squid heart muscle cells, and that the underlying G protein pathway is dissimilar to that commonly associated with transmitter modulation of channel activity. The FMRFamide modulated increase in ICa,L seen in the type II heart muscle cells was not mediated by a PTX-sensitive G protein pathway. PMID- 10835050 TI - Voltage- and ligand-gated ryanodine receptors are functionally separated in developing C2C12 mouse myotubes. AB - In order to further understand the role of voltage- and ligand-gated ryanodine receptors in the control of intracellular Ca2+ signalling during myogenesis, changes in cytosolic free calcium concentration ([Ca2+]i) were investigated by fura-2 videoimaging in C2C12 mouse myotubes developing in vitro. A synchronous [Ca2+]i increase was observed after depolarisation with high [K+], while the Ca2+ response propagated as a wave following caffeine administration. Application of the two stimuli to the same myotube often revealed the existence of cellular zones that were responsive to depolarisation but not to caffeine. Focal application of high [K+] promoted a [Ca2+]i response detectable only in the cellular areas close to the pipette tip, while focal application of caffeine elicited a [Ca2+]i increase which spread as a Ca2+ wave. Buffering of [Ca2+]i by BAPTA did not affect the pattern of the depolarisation-induced [Ca2+]i transient but abolished the Ca2+ waves elicited by caffeine. When high [K+] and caffeine were applied in sequence, reciprocal inhibition of the [Ca2+]i responses was observed. Our results suggest that the different spatial patterns of [Ca2+]i responses are due to uneven distribution of voltage- and ligand-gated ryanodine receptors within the myotube. These two types of receptor control two functionally distinct Ca2+ pools which are part of a common intracellular compartment. Finally, the two differently operated ryanodine receptor channels appear to be independently activated, so that a mechanism of Ca2+-induced Ca2+ release is not required to sustain the global response in C2C12 myotubes. PMID- 10835049 TI - Remodelling of ionic currents in hypertrophied and failing hearts of transgenic mice overexpressing calsequestrin. AB - Overexpression of cardiac calsequestrin (CSQ) impairs Ca2+ signalling in murine myocytes, leading to marked cardiac hypertrophy. Here we report on contractile, histological and electrophysiological changes accompanying the development of cardiac hypertrophy and failure in CSQ-overexpressing mice. CSQ mice developed contractile dysfunction after 60 days of age, with only 40% survival at 6 months. Four- to 6-month-old CSQ mice revealed biventricular dilatation, cardiomyocyte hypertrophy, patchy interstitial fibrosis and tissue calcifications. Cardiac hypertrophy of CSQ mice was accompanied by progressive P-R and Q-T interval prolongation, conduction blocks, 2-fold prolongation of the ventricular action potential and increased cellular membrane capacitance. Remodelling of ionic currents included marked reduction of both density and absolute magnitude of transient outward (Ito) and inward rectifying (IK1) K+ currents. The density, but not the absolute magnitude, of basal and isoproterenol (isoprenaline)-stimulated Ca2+ current (ICa) was decreased by 42% and the inactivation kinetics of ICa were significantly slowed. Na+ current density was suppressed by 50%, but its steady state activation and inactivation were shifted to more positive potentials. The density of Na+-Ca2+ exchange current was increased by 35%. In CSQ but not in control myocytes dialysed with cAMP, isoproterenol continued to enhance ICa. This apparent lower responsiveness of ICa to cAMP could be reversed by the non hydrolysable cAMP analogue 8-Br-cAMP or the phosphodiesterase inhibitor IBMX, suggesting high phosphodiesterase activity of CSQ myocytes. In young CSQ mice (< 60 days) with compensated cardiac hypertrophy, only Ito was significantly suppressed. All other currents remained relatively intact. An increase in cardiac Ca2+-storage capability by overexpression of CSQ results in a dilated cardiomyopathy with tissue fibrosis, calcifications, impaired beta-adrenergic signalling and progressive remodelling of ionic currents. The extent of the changes in ionic currents was age dependent. PMID- 10835051 TI - Transient configurations of baroresponsive respiratory-related brainstem neuronal assemblies in the cat. AB - The regulation of gas exchange requires coordination of the respiratory and cardiovascular systems. Previous work suggested that medullary raphe neurones transform and transmit information from baroreceptors to neurones in the ventral respiratory group. This study tested the hypothesis that distributed brainstem neuronal assemblies are transiently reconfigured during the respiratory cycle and baroreceptor stimulation. Blood pressure was perturbed by intravenous injection of an alpha1-adrenergic receptor agonist, unilateral pressure changes in the carotid sinus, or occlusion of the descending aorta in 14 Dial-urethane anaesthetized, vagotomized, paralysed, artificially ventilated cats. Neurones were monitored simultaneously with microelectrode arrays in two or more of the following sites: n. raphe obscurus, n. raphe magnus, rostral and caudal ventrolateral medulla, and the nucleus tractus solitarii. Transient configurations of baroresponsive assemblies were detected with joint pericycle triggered histograms, the gravitational representation, and related pattern detection methods. Data were also analysed with cycle-triggered histograms, peristimulus-time and cumulative sum histograms, cross-correlograms, spike triggered averages of efferent phrenic activity, and joint impulse configuration scatter diagrams (snowflakes). Five to nine simultaneously recorded spike trains from control expiratory phases were compared with data from interleaved equal duration time blocks from control inspiratory phases. In each of seven animals, significant impulse synchrony detected by gravity analysis was confined to one phase of the respiratory cycle. Repeated patterns of distributed synchrony confined to periods of altered baroreceptor activity were detected and involved neurones that individually did not change firing rates during stimulation. Snowflakes and logical cross-correlation analysis provided evidence for the cooperative actions of impulses in concurrently active parallel channels. In 12 of 17 pairs of neurones with at least one baroresponsive cell, joint pericycle triggered histograms detected synchrony indicative of shared inputs or functional excitatory interactions that varied as a function of time in the respiratory cycle. Neurones in four of the pairs had no respiratory modulation of their individual firing rates. Data from eight other pairs were indicative of fluctuations in inhibition during the respiratory cycle. The results demonstrate repeated transient configurations of baroresponsive neuronal assemblies during the respiratory cycle, without concomitant firing rate changes in the constituent neurones, and suggest distributed network mechanisms for the modulation of baroreceptor-mediated reflexes. PMID- 10835052 TI - A thixotropic effect in contracting rabbit psoas muscle: prior movement reduces the initial tension response to stretch. AB - Paired ramp stretches and releases ('triangular length changes', typically 0.04 +/- 0.09L0 s-1; mean +/- s.e.m.) were imposed on permeabilised rabbit psoas fibre segments under sarcomere length control. In actively contracting fibres, the tension response to stretch was biphasic; tension rose more rapidly during the first 0. 005L0 of the imposed stretch than thereafter. Tension also dropped in a biphasic manner during shortening, and at the end of the length change was reduced below the steady state. If a second triangular length change was imposed shortly after the first, tension rose less sharply during the initial phase of lengthening, i.e. the stiffness of the muscle during the initial phase of the response was reduced in the second stretch. This is a thixotropic effect. If a third triangular length change was imposed on the muscle, the response was the same as that to the second. The time required to recover the original tension response was measured by varying the interval between triangular length changes. Recovery to steady state occurred at a rate of approximately 1 s-1. The stiffness of the muscle during the initial phase of the response scaled with the developed tension in pCa (= -log10[Ca2+]) solutions ranging from 6.3 (minimal activation) to 4.5 (saturating effect). The relative thixotropic reduction in stiffness measured using paired length changes was independent of the pCa of the activating solution. The thixotropic behaviour of contracting skeletal muscle can be explained by a cross-bridge model of muscle contraction in which the number of attached cross-bridges is temporarily reduced following an imposed movement. PMID- 10835054 TI - The danger of premature graduation to seat belts for young children. AB - OBJECTIVE: To determine the risk of significant injury associated with premature graduation of young (2- to 5-year-old) children to seat belts from child restraint systems (CRS). BACKGROUND: Advocates recommend use of child safety seats for children younger than age 4 and booster seats for children age 4 and older. Despite these recommendations, many children are prematurely taken out of these child restraints and placed in seat belts. Although data exist to support the use of child restraints over nonrestraint, no real-world data exist to evaluate the risk of significant injury associated with premature use of seat belts. DESIGN/METHODS: Partners for Child Passenger Safety includes a child focused crash surveillance system based on a representative sample of children ages 0 to 15 years in crashes involving 1990 and newer vehicles reported to State Farm Insurance Companies in 15 states and the District of Columbia. Driver reports of crash circumstances and parent reports of child occupant injury were collected via telephone interview using validated surveys. Results were weighted based on sampling frequencies to represent the entire population. RESULTS: Between December 1, 1998, and November 30, 1999, 2077 children aged 2 to 5 years were included and were weighted to represent 13 853 children. Among these young children, 98% were restrained, but nearly 40% of these children were restrained in seat belts. Compared with children in CRS, children in seat belts were more likely to suffer a significant injury (relative risk: 3.5; 95% confidence interval: [2. 4, 5.2]). Children in seat belts were at particular risk of significant head injuries (relative risk: 4.2; 95% confidence interval: [2.6, 6.7]) when compared with children in CRS. CONCLUSIONS: Premature graduation of young children from CRS to seat belts puts them at greatly increased risk of injury in crashes. A major benefit of CRS is a reduction in head injuries, potentially attributable to a reduction in the amount of head excursion in a crash. PMID- 10835053 TI - Group I disynaptic excitation of cat hindlimb flexor and bifunctional motoneurones during fictive locomotion. AB - The incidence of short latency excitation of motoneurones innervating flexor and bifunctional muscles evoked by group I intensity (<= 2 x threshold) electrical stimulation of hindlimb muscle nerves was investigated during fictive locomotion in decerebrate cats. Intracellular recordings were made from hindlimb motoneurones in which action potentials were blocked by intracellular diffusion of a lidocaine (lignocaine) derivative (QX-314) and fictive locomotion was evoked by electrical stimulation of the midbrain. Few motoneurones (16%) received group I-evoked oligosynaptic excitation in the absence of fictive locomotion. During fictive locomotion 39/44 (89%) motoneurones innervating ankle, knee or hip flexor muscles and 18/28 (64%) motoneurones innervating bifunctional muscles received group I-evoked oligosynaptic EPSPs. In flexor motoneurones, locomotor-dependent excitation was present in both step cycle phases but largest during flexion. In bifunctional motoneurones, EPSPs were often largest at the transition between flexion and extension phases. Activation of homonymous afferents most consistently evoked the largest locomotor-dependent excitation (amplitude up to 4.6 mV), but in some cases stimulation of heteronymous flexor or bifunctional muscle nerves evoked large EPSPs. EPSP amplitude became maximal as stimulation intensity was increased to about twice threshold. This suggests that tendon organ afferents can evoke group I EPSPs during locomotion. The EPSPs resulting from brief, small stretches of extensor digitorum longus tendons indicate that group Ia muscle spindle afferents can also evoke the group I excitation of flexors. Stimulation of extensor group I afferents did not result in excitation of flexor motoneurones. The mean latency of locomotor-dependent group I excitation in flexor and bifunctional motoneurones was 1.64 +/- 0.16 ms, indicating a path consisting of a single interneurone interposed between group I afferents and motoneurones innervating flexor and bifunctional muscles. This disynaptic excitation is analogous to that recorded in extensor motoneurones and evoked from extensor group I afferents during locomotion. Differences in the phase dependence and sources of group I excitation to flexor and extensor motoneurones during locomotion suggest the existence of separate groups of excitatory interneurones exciting flexor and extensor motoneurones. The wide distribution of group I disynaptic excitation in motoneurones innervating extensor, flexor and bifunctional muscles acting on hip, knee and ankle joints suggests that these pathways can play an important role in the reinforcement of ongoing locomotor activity throughout the limb. PMID- 10835055 TI - Clinical auscultation skills in pediatric residents. AB - OBJECTIVE: The aim of this study is to determine the level of clinical auscultation skills in pediatric residents at Duke University Medical Center. METHODS: Forty-seven residents from pediatrics and joint medicine/pediatrics training programs at Duke University Medical Center were enrolled in this study. They were asked to examine the cardiovascular patient simulator, Harvey, and report their findings. Five common conditions seen in the pediatric population were presented: ventricular septal defect, atrial septal defect, pulmonary valve stenosis, combined aortic valve stenosis and insufficiency, and innocent systolic ejection murmur. The responses were scored by the number of features and diagnoses accurately reported. Five pediatric cardiologists and cardiologists in training were also asked to participate in a manner similar to the trainees. RESULTS: The mean score of features identified for the resident group was 11.4 +/ 2.6 of a possible 19. The diagnostic accuracy was 33%. There was no significant difference between residents by year of training or by type of residency program, although there was a trend toward improved performance with more training. The difference in performance between the pediatric cardiology group and the residents group was striking. The condition that was most frequently misdiagnosed was the innocent systolic ejection murmur. CONCLUSIONS: The clinical auscultation skills of pediatric residents in this study were suboptimal. There was a trend toward improvement as training progressed, although not statistically significant. These skills are likely to improve further with increased exposure to patients with cardiovascular disease especially in the ambulatory care setting. PMID- 10835056 TI - Medication use among children with asthma in East Harlem. AB - PURPOSE: To examine daily use of antiinflammatory medication among children with asthma in East Harlem, where hospitalization rates for asthma are among the highest in the United States. METHODS: We analyzed parent/guardian reports of medications used by children with current asthma (defined as physician diagnosis and wheezing during the previous 12 months) identified from a cross-sectional survey conducted in 2 elementary schools. RESULTS: From an overall sample of 1319 children, 298 with current asthma were included in this analysis. Most of those with asthma were Puerto Rican (136 [46%]) or black (98 [33%]), 168 (57%) were boys, and the median age was 8 years old. Overall, 65 (22%) were using antiinflammatory medication on a daily basis. A subgroup of 107 children with asthma had been hospitalized during the previous 12 months or had used beta(2) agonist on a daily basis, suggesting persistent or severe asthma. Of these 107 children, 42 (39%) were taking antiinflammatory medication on a daily basis. Multivariate analysis of these 107 children revealed that daily use of antiinflammatory medication was associated with using a spacer tube (adjusted odds ratio [AOR]: 3. 08; 95% confidence interval [CI]: 1.27,7.47) and having seen a physician in the past 6 months (AOR: 3.46; CI: 1.01,11.9). Compared with Puerto Ricans, blacks (AOR:.32; CI:.12,.89) or children of other races/ethnicities (AOR:.27; CI:.09,.85) were less likely to use antiinflammatory medication on a daily basis. CONCLUSION: Daily use of antiinflammatory medication for children with persistent or severe asthma in East Harlem was underused. Differences in access to care may explain some findings; however, reasons for ethnic differences in use remain unclear. Both community interventions and additional provider education are needed. PMID- 10835057 TI - Do clinical markers of barotrauma and oxygen toxicity explain interhospital variation in rates of chronic lung disease? The Neonatology Committee for the Developmental Network. AB - OBJECTIVE: To explore the hypothesis that variation in respiratory management among newborn intensive care units (NICUs) explains differences in chronic lung disease (CLD) rates. DESIGN: Case-cohort study. SETTING: NICUs at 1 medical center in New York (Babies' and Children's Hospital [Babies']) and 2 in Boston (Beth Israel Hospital and Brigham and Women's Hospital [Boston]). STUDY POPULATION: Four hundred fifty-two infants born at 500 to 1500 g birth weight between January 1991 and December 1993, who were enrolled in an epidemiologic study of neonatal intracranial white matter disorders. CASE DEFINITION: Supplemental oxygen required at 36 weeks' postmenstrual age. RESULTS: The prevalence rates of CLD differed substantially between the centers: 4% at Babies' and 22% at the 2 Boston hospitals, despite similar mortality rates. Initial respiratory management at Boston was more likely than at Babies' to include mechanical ventilation (75% vs 29%) and surfactant treatment (45% vs 10%). Case and control infants at Babies' were more likely than were those at Boston to have higher partial pressure of carbon dioxide and lower pH values on arterial blood gases. However, measures of oxygenation and ventilator settings among case and control infants were similar at the 2 medical centers in time-oriented logistic regression analyses. In multivariate logistic regression analyses, the initiation of mechanical ventilation was associated with increased risk of CLD: after adjusting for other potential confounding factors, the odds ratios for mechanical ventilation were 13.4 on day of birth, 9.6 on days 1 to 3, and 6.3 on days 4 to 7. Among ventilated infants, CLD risk was elevated for maximum peak inspiratory pressure >25 and maximum fraction of inspired oxygen = 1.0 on the day of birth, lowest peak inspiratory pressure >20 and maximum partial pressure of carbon dioxide >50 on days 1 to 3, and lowest white blood count <8 K on days 4 to 7. Even after adjusting for white blood count <8 K and the 4 respiratory care variables, infants in Boston continued to be at increased risk of CLD, compared with premature infants at Babies' Hospital. CONCLUSION: In multivariate analyses, a number of specific measures of respiratory care practice during the first postnatal week were associated with the risk of a very low birth weight infant developing CLD. However, after adjusting for baseline risk, most of the increased risk of CLD among very low birth weight infants hospitalized at 2 Boston NICUs, compared with those at Babies' Hospital, was explained simply by the initiation of mechanical ventilation. PMID- 10835058 TI - First intention high-frequency oscillation with early lung volume optimization improves pulmonary outcome in very low birth weight infants with respiratory distress syndrome. AB - OBJECTIVES: The lack of decline in chronic lung disease of prematurity despite the generalized use of surfactant and alternative modes of ventilation such as high-frequency oscillation (HFO) has been attributed to some misunderstanding of how HFO has to be used. We used a new ventilatory strategy in very low birth weight (VLBW) infants, by initiating HFO immediately after intubation and attempting early lung volume optimization before surfactant was administered. STUDY DESIGN: The outcome of 32 VLBW infants, managed with first intention HFO over a period of 24 months (September 1, 1996 and August 31, 1998) was compared by chart review with 39 historical controls, consecutively managed with conventional mechanical ventilation (CMV) over a period of 24 months (January 1, 1994 and December 31, 1995). SETTING: An 11-bed tertiary care pediatric and neonatal intensive care unit of a university teaching hospital. RESULTS: The 2 groups of patients were similar in demographic distribution of birth weight, gestational age, race, and gender. Patients on first intention HFO were ventilator-dependent (median [95% confidence interval]: 5 [3-6] vs 14 [6-23] days) and oxygen-dependent (12 [4-17] vs 51 [20-60] days) for a shorter time than patients on CMV. The incidence of chronic lung disease at 36 weeks of gestational age was significantly lower in the HFO group compared with the CMV group (0% vs 34%). CONCLUSIONS: First intention HFO with early lung volume optimization shortened the need for respiratory support and improved pulmonary outcome of VLBW infants with respiratory distress syndrome significantly. PMID- 10835059 TI - Pulmonary outcome in extremely low birth weight infants. AB - OBJECTIVE: To determine whether infants with hyaline membrane disease (HMD) superimposed on immature lung disease (ILD) have more abnormal lung function and respiratory drive during the evolution of chronic neonatal lung disease (CNLD) in extremely low birth weight infants (ELBW; <1000 g). METHODS: We measured lung mechanics (respiratory frequency, tidal volume, minute ventilation, lung resistance, lung compliance, lung impedance, and work of breathing per minute) and respiratory drive (airway opening pressure 100 milliseconds after initiation of breath [P(0.1)] and maximal inspiratory pressure generated during airway occlusion) on 3 occasions before term in 24 ELBW infants. RESULTS: Ten infants with ILD (mean [95% CI] gestation: 24.3 weeks [23.1,25.4]; birth weight: 675 g [553,798]) were studied at 27, 31, and 35 weeks of postconceptional age and 14 infants with HMD superimposed on ILD (gestation: 25.1 weeks [24.4,25.9]; birth weight: 687 g [601,773]) were studied at 28, 32, and 35 weeks of postconceptional age. There were no statistically significant differences between the groups for respiratory frequency, tidal volume, minute ventilation, lung resistance, lung compliance, lung impedance, work of breathing per minute, P(0.1), and maximal inspiratory pressure generated during airway occlusion. With increasing age, both groups demonstrated increased respiratory drive as measured by P(0.1) without significant changes in respiratory frequency or CO(2). Work of breathing per minute increased in the HMD group with age and was higher in extubated subjects. A similar trend with age was demonstrated in ILD infants. Regardless of whether the initial lung disease was ILD alone or HMD + ILD, ELBW infants developed a mildly reduced lung compliance/kg (.8-1.1 mL/cm.H(2)O/kg) and high lung resistance (75-125 cm.H(2)O/L/second) pattern of CNLD, which changed little after 3 weeks of age. Survival to 6 months was 23/24 (96%). Oxygen dependency was 16/24 (67%) at 35 weeks, yet only 5/23 (22%) survivors required oxygen at discharge from the neonatal unit (43 weeks). CONCLUSIONS: The visco-elastic and flow resistive properties of the lungs in ELBW infants with CNLD remain only mildly abnormal, suggesting a more favorable prognosis for lung function in later years than previously reported. PMID- 10835060 TI - Neurodevelopmental and functional outcomes of extremely low birth weight infants in the National Institute of Child Health and Human Development Neonatal Research Network, 1993-1994. AB - OBJECTIVES: The purposes of this study were to report the neurodevelopmental, neurosensory, and functional outcomes of 1151 extremely low birth weight (401 1000 g) survivors cared for in the 12 participating centers of the National Institute of Child Health and Human Development Neonatal Research Network, and to identify medical, social, and environmental factors associated with these outcomes. STUDY DESIGN: A multicenter cohort study in which surviving extremely low birth weight infants born in 1993 and 1994 underwent neurodevelopmental, neurosensory, and functional assessment at 18 to 22 months' corrected age. Data regarding pregnancy and neonatal outcome were collected prospectively. Socioeconomic status and a detailed interim medical history were obtained at the time of the assessment. Logistic regression models were used to identify maternal and neonatal risk factors for poor neurodevelopmental outcome. RESULTS: Of the 1480 infants alive at 18 months of age, 1151 (78%) were evaluated. Study characteristics included a mean birth weight of 796 +/- 135 g, mean gestation (best obstetric dates) 26 +/- 2 weeks, and 47% male. Birth weight distributions of infants included 15 infants at 401 to 500 g; 94 at 501 to 600 g; 208 at 601 to 700 g; 237 at 701 to 800 g; 290 at 801 to 900 g; and 307 at 901 to 1000 g. Twenty five percent of the children had an abnormal neurologic examination, 37% had a Bayley II Mental Developmental Index <70, 29% had a Psychomotor Developmental Index <70, 9% had vision impairment, and 11% had hearing impairment. Neurologic, developmental, neurosensory, and functional morbidities increased with decreasing birth weight. Factors significantly associated with increased neurodevelopmental morbidity included chronic lung disease, grades 3 to 4 intraventricular hemorrhage/periventricular leukomalacia, steroids for chronic lung disease, necrotizing enterocolitis, and male gender. Factors significantly associated with decreased morbidity included increased birth weight, female gender, higher maternal education, and white race. CONCLUSION: ELBW infants are at significant risk of neurologic abnormalities, developmental delays, and functional delays at 18 to 22 months' corrected age. PMID- 10835061 TI - Birth weight- and gestational age-specific sudden infant death syndrome mortality: United States, 1991 versus 1995. AB - OBJECTIVE: To estimate the changes in birth weight- and gestational age-specific sudden infant death syndrome (SIDS) mortality rates since the publication of the sleep-positioning recommendations by the American Academy of Pediatrics Task Force on Infant Positioning and SIDS. METHODS: This is a historical cohort study using US vital statistic linked birth and infant death certificate files for the years 1991 and 1995. SIDS deaths were identified as any death attributed to International Classification of Diseases, Ninth Revision code 7980, occurring between the 28th and 365th days of life. RESULTS: There were 4871 deaths attributed to SIDS in 1991 for a postneonatal mortality rate of 1.2/1000 postneonatal survivors compared with 3114 deaths in 1995 for a rate of.8/1000. This represents a 33% drop in the postneonatal SIDS mortality from 1991 to 1995. Between 1991 and 1995, SIDS rates declined 38%, 38%, 35%, and 32% for birth weight groupings of 500 to 999 g, 1000 to 1499 g, 1500 to 2499 g, and >/=2500 g, respectively. There were no SIDS deaths attributed to infants weighing <500 g. The SIDS rates declined 27%, 21%, 40%, and 23% for gestational age groups of <29 weeks, 29 to 32 weeks, 33 to 36 weeks, and >/=37 weeks. The rate of decline did not differ significantly across birth weight- or gestational age-specific categories. There was a significant increase in the black:non-black postneonatal SIDS mortality ratio from 2.00 to 2.28, reflecting a smaller decline in birth weight- and gestational age-specific mortality for blacks than observed for the non-black population. CONCLUSION: Postneonatal SIDS mortality decreased significantly across all broad birth weight and gestational age categories. If the decline in the prevalence of prone positioning that has been reported since 1992 has occurred across all birth weight and gestational age, these data support the hypothesis that supine or side sleep positioning is effective in preterm/low birth weight infants as well as term infants. PMID- 10835062 TI - Imaging after urinary tract infection in male neonates. AB - OBJECTIVE: To assess the frequency of urinary tract anomalies in male neonates <8 weeks old who presented with urinary tract infection (UTI), and to evaluate a suitable imaging approach after the initial infection. DESIGN: During a period of 4.5 years, from July 1994 through December 1998, 45 male neonates <8 weeks old (range: 5-56 days; mean: 23.77 days) with UTI were hospitalized. All patients had an ultrasound (US) and a voiding cystourethrogram (VCUG), except 1 neonate in whom VCUG was unsuccessful because of technical problems. A dimercaptosuccinic acid (DMSA) scan was recommended to all patients but was performed only in 30 of 45, most of them with an abnormal VCUG. The renal scan was performed at least 4 months after the UTI. RESULTS: Urinary tract abnormalities were observed in 22 of 45 male neonates. Nineteen had vesicoureteral reflux (VUR), 1 had VUR and a double collecting system, 1 had VUR and a posterior urethral valve, and 1 had an ureteropelvic junction stricture. Renal atrophy or scars, as demonstrated by DMSA scan, were detected almost exclusively in neonates with VUR grade 3 and above. Only 1 neonate with VUR grade 1 had a pathologic DMSA, and the US of this male also demonstrated renal atrophy. Escherichia coli was the pathogen in 62% (28 of 45), and 9 boys had bacteremia. CONCLUSION: We suggest that US and VCUG should be performed routinely after the initial UTI in male neonates. Renal scan should be reserved for those cases in which the US suggests renal parenchymal damage or when VCUG detects VUR grade 3 and above. PMID- 10835063 TI - Does treatment of vesicoureteric reflux in childhood prevent end-stage renal disease attributable to reflux nephropathy? AB - OBJECTIVE: It is believed that end-stage renal disease (ESRD) attributable to reflux nephropathy is preventable by the active treatment of vesicoureteric reflux in childhood with long-term antibiotics and ureteric reimplantation surgery. We aimed to test this belief. METHODOLOGY: The Australia and New Zealand Dialysis and Transplant Registry of new patients 5 to 44 years of age treated for ESRD between 1971 and 1998, categorized by age and primary renal disease, was used to analyze the age-specific incidences of ESRD attributable to reflux nephropathy using a before-after study design. The early 1960s were regarded as the introduction period for the active treatment of childhood vesicoureteric reflux. A time-delay in treatment effect was expected. Patients with ESRD attributable to other causes were used as a comparative group. RESULTS: The incidence of ESRD attributable to reflux nephropathy and nonreflux nephropathy has increased. For reflux nephropathy, the rate of change was significantly associated with age, with a downward trend in incidence with decreasing age suggesting a minor treatment effect. This trend was no longer evident when adjustment was made for changing diagnostic practices. An opposite trend was observed for the nonreflux nephropathy group, who demonstrated an upward trend in incidence with decreasing age. CONCLUSIONS: Treatment of children with vesicoureteric reflux has not been accompanied by the hoped-for reduction in the incidence of ESRD attributable to reflux nephropathy. A randomized trial with a control (no-treatment) arm is required to appropriately assess the medical belief that long-term antibiotics and surgery improve the natural history of vesicoureteric reflux. PMID- 10835064 TI - Evaluation and management of proteinuria and nephrotic syndrome in children: recommendations from a pediatric nephrology panel established at the National Kidney Foundation conference on proteinuria, albuminuria, risk, assessment, detection, and elimination (PARADE). AB - OBJECTIVE: The development of this review article evolved from a National Kidney Foundation consensus conference on recent advances in the importance of evaluating and treating proteinuria. From this conference, a series of recommendations for the evaluation of adults with proteinuria was published. Because specific pediatric aspects of the problem were outside the scope of the original National Kidney Foundation publication, an ad hoc committee of 6 pediatric nephrologists who were active participants in the National Kidney Foundation conference was established to provide primary care physicians with a concise, up-to-date reference on this subject. METHODS: The recommendations that are given represent the consensus opinions of the authors. These are based on data from controlled studies in children when available, but many of the opinions are, by necessity, based on uncontrolled series in children or controlled trials performed in adults, because controlled trials in children have not been performed to evaluate many of the treatments described. RESULTS AND CONCLUSIONS: These recommendations are intended to provide primary care physicians with a useful reference when they are faced with a young child or teenager who presents with proteinuria, whether this is mild and asymptomatic or more severe, leading to nephrotic syndrome. PMID- 10835065 TI - Changing indications for pediatric cholecystectomy. AB - PURPOSE: The purpose of this study was to determine indications for gallbladder surgery and risk factors for urgent surgery. METHODS: We reviewed all patients <19 years old, who underwent cholecystectomy between 1980 and 1996. RESULTS: There were 128 patients (mean age: 10 years). Fifty-two patients had an underlying hematologic disorder, 47 had another medical disorder, and 29 had no preexisting illness or identifiable risk factor for gallstone disease. Twenty five percent (32/128) of cholecystectomies were performed urgently. Postoperative complications developed in 5 of 32 patients (16%) who underwent emergency surgery and 6 of 96 patients (6%) who underwent elective surgery. There were 3 deaths, all occurring in patients undergoing emergency cholecystectomy (odds ratio: 23). Furthermore, all who died had congenital heart disease (odds ratio: 183), making congenital heart disease an independent risk factor for gallstone-related mortality. CONCLUSIONS: Cholecystectomy is recommended when medically possible for children with underlying medical diseases. Patients with medical disorders that make them a high surgical risk can be followed clinically, realizing that if urgent surgery is necessary, the morbidity is relatively high. Those children with congenital heart disease and gallstones are at a prohibitively high risk for death after urgent cholecystectomy. For these patients, the risk of an elective cholecystectomy may be acceptable when weighed against the high risk of complications from their gallstones. PMID- 10835066 TI - Screening for iron deficiency anemia by dietary history in a high-risk population. AB - BACKGROUND: Iron deficiency anemia (IDA) in young children is important to identify because of its adverse effects on behavior and development. Because of costs and inconvenience associated with blood test screening and the decline in prevalence of IDA, the Institute of Medicine and the Centers for Disease Control and Prevention recommend that blood test screening for IDA be targeted to children first identified by dietary and health history. OBJECTIVE: To evaluate a parent-completed dietary and health history as the first stage of 2-stage screening for IDA. DESIGN AND METHODS: A cross-sectional study was conducted in inner-city clinics in children 9 to 30 months old having routine anemia screening as part of a scheduled visit. Parents completed a questionnaire and children had venous blood sampling for complete blood count and ferritin. Anemia was defined as Hb <11.0 g/dL. Iron deficiency (ID) was defined as ferritin <10 microg/L or mean corpuscular volume <70 fL and red cell distribution width >14.5%. Children were categorized into 1 of 4 groups: iron-sufficient, not anemic (ISNA); iron sufficient, anemic (ISA); iron-deficient, not anemic (IDNA); and iron-deficient anemic (IDA). The questionnaire consisted of 15 dietary items in domains of infant diet, intake of solid food, intake of beverages, and participation in the Special Supplemental Nutrition Program for Women, Infants, and Children together with 14 historical items in domains of birth history, recent illness, chronic medical conditions, history of anemia, and maternal history. Analysis was performed on individual items, domains, and combinations of selected items. RESULTS: In the 282 study subjects, the prevalence of anemia (35%), IDNA (7%), and IDA (8%) did not vary significantly by age. Among individual historical and dietary questions, maternal history of anemia and drinking >2 glasses of juice per day identified the highest proportion of children with IDA: 50% sensitivity (95% confidence interval [CI]: 16,81) and 77% sensitivity (95% CI: 54,89), respectively. However, specificities for these questions were 60% (95% CI: 55,65) and 22% (95% CI: 17,27), respectively. Domains of questions with the highest sensitivity for IDA were beverage intake (91%; 95% CI: 68,99) and intake of solid food (91%; 95% CI: 68,99). However, specificities of the domains were only 14% (95% CI: 10,18) and 29% (95% CI: 24,35), respectively. The dietary items used by Boutry and Needlman were 95% (95% CI: 77, 99) sensitive but only 15% (95% CI: 11,19) specific for IDA. The recommendations of the Centers for Disease Control and Prevention for health and dietary screening were 73% (95% CI: 56,92) sensitive and 29% (95% CI: 24,35) specific for IDA. The individual questions, domains of questions, and interdomain groups of questions had similar sensitivity and specificity for anemia and ID (IDA + IDNA). CONCLUSION: In this high-risk population, neither individual nor combinations of parental answers to dietary and health questions were able to predict IDA, anemia, or ID well enough to serve as a first-stage screening test. PMID- 10835067 TI - Metabolic aspects of myocardial disease and a role for L-carnitine in the treatment of childhood cardiomyopathy. AB - OBJECTIVES: A multicenter retrospective study was conducted to investigate the possible metabolic causes of pediatric cardiomyopathy and evaluate the outcome of patients treated with L-carnitine. METHODS: Seventy-six patients diagnosed with cardiomyopathy were treated with L-carnitine in addition to conventional cardiac treatment, and 145 patients were treated with conventional treatment only. There were 101 males and 120 females between 1 day and 18 years old. Cardiomyopathy diagnoses included dilated (148 patients), hypertrophic (42 patients), restrictive (16 patients), mixed diagnosis (11 patients), and 4 with an unknown type. Of 76 L-carnitine-treated patients, 29 (38%) had evidence to suggest a disorder of metabolism, and of 145 control patients, 15 (10%) were suspected to have a disorder of metabolism. These metabolic disorders were thought to be the cause for the cardiomyopathy of the patients. The duration of L-carnitine treatment ranged from 2 weeks to >1 year. Information was collected on length of survival (time-to-event), clinical outcome, echocardiogram parameters, and clinical assessments. Data were collected at intervals from baseline to study endpoint, death, transplant, or last known follow-up visit. RESULTS: L-Carnitine treated patients were younger than control patients and had poorer clinical functioning at baseline, yet they demonstrated lower mortality and a level of clinical functioning and clinical severity comparable to control patients on conventional therapy by the end of the study. An analysis of the interaction between clinical outcome and concomitant medications unexpectedly revealed that the population of patients treated with angiotensin-converting enzyme (ACE) inhibitors (40% of patients) had significantly poorer survival (although their greater likelihood for poor survival may possibly have made them more likely to receive ACE inhibitors). CONCLUSION: Results suggest that L-carnitine provides clinical benefit in treating pediatric cardiomyopathy. There is a need for further exploration of potential explanatory factors for the higher mortality observed in the population of patients treated with ACE inhibitors. PMID- 10835068 TI - The impact of managed care insurance on use of lower-mortality hospitals by children undergoing cardiac surgery in California. AB - CONTEXT: Managed care plans aggressively seek to contain costs, but few data are available regarding their impact on access to high quality care for their members. OBJECTIVE: To assess the impact of managed care health insurance on use of lower-mortality hospitals for children undergoing heart surgery in California. DESIGN: Retrospective cohort study using state-mandated hospital discharge datasets. SETTING: Pediatric cardiovascular surgical centers in California. PATIENTS: Five thousand seventy-one children admitted for open cardiac surgical procedures during 1992-1994. RESULTS: Hospitals were divided into lower- and higher-mortality groups according to adjusted surgical mortality. Using multivariate logistic regression analysis to control for medical, socioeconomic, demographic, and distance factors, children with managed care insurance were less likely to be admitted to a lower-mortality hospital for surgery relative to children with indemnity insurance (odds ratio:.53; 95% confidence interval:.45,.63). Similar findings resulted when the analysis was stratified by race/ethnicity. In addition, length of stay, a correlate of health care costs, was no longer for children admitted to lower-mortality centers than for those at higher-mortality centers (adjusted difference:.54 days shorter at lower-mortality centers; 95% confidence interval: -1.50,. 41). CONCLUSIONS: During this study, children with managed care insurance had significantly reduced use of lower mortality hospitals for pediatric heart surgery in California compared with children with indemnity insurance. Further study is necessary to determine the mechanisms of this apparent insurance-specific inequity. PMID- 10835070 TI - Thrombopoietin in the thrombocytopenic term and preterm newborn. AB - OBJECTIVES: Neonatal thrombocytopenia occurs commonly in neonatal intensive care units. The role of the thrombopoietin (Tpo) system in normal neonatal platelet regulation and neonatal thrombocytopenia is not well understood. The purpose of our study was to: 1) determine the normal Tpo level at birth in healthy nonthrombocytopenic term (NTT) and nonthrombocytopenic preterm (NTP) infants and in infants born to women with preeclampsia; and 2) measure Tpo levels in infants during and after the resolution of thrombocytopenia. Characterizing Tpo levels in the healthy and thrombocytopenic newborn is an important step in furthering our understanding of the pathophysiology of neonatal thrombocytopenia. METHODS: This study is comprised of 2 parts. For the first part, cord blood was obtained at birth from both term (gestational age [GA]: 38-42 weeks) and preterm (GA: 25-36 weeks) infants. If birth platelet levels were >/=140 x 10(3)/microL and the infant fit criteria for being normal, or if the infant was born to a women with preeclampsia, Tpo levels were measured. For the second part, serial Tpo levels and concomitant platelet counts (Plts) were measured in both preterm and term infants during a period of marked thromboctyopenia (Plt < 100 x 10(3)/microL) until its resolution (Plt >/= 140 x 10(3)/microL). RESULTS: Median cord blood Tpo levels from NTP infants (n = 35) were higher than those of NTT infants (n = 32; 95 pg/mL vs 48 pg/mL, respectively). In addition, preterm infants born to women with preeclampsia (n = 11) had lower Tpo levels than NTP infants with a similar GA (<41 pg/mL vs 95 pg/mL). For infants with marked thrombocytopenia, median Tpo levels during thrombocytopenia were similar between term (n = 12) and preterm (n = 14) groups (223 pg/mL and 179 pg/mL, respectively), with the majority of individuals showing a decrease in Tpo with resolution of thrombocytopenia. Within each group, there was large variability in the Tpo response to thrombocytopenia. IMPRESSION: These data show that the Tpo system is intact in NTP and NTT neonates. Preeclampsia may be an example of a disorder that perturbs this system. The great variability in Tpo levels seen in infants during thrombocytopenia may be related to the mechanism of thrombocytopenia. The finding that, in general, Tpo levels decreased with resolution of thrombocytopenia is consistent with what has been described in adults and children. PMID- 10835069 TI - Effect of neonatal test environment on recording transient-evoked otoacoustic emissions. AB - OBJECTIVE: To examine the effect of test environment on recording transient evoked otoacoustic emissions (TEOAE) in neonates. METHODS: Thirty-two infants from the newborn nursery (NBN) who passed a screening auditory brainstem response (SABR) test and were at least 34 weeks' postconceptional age were studied. One ear of each newborn was tested using TEOAE in 5 different test environments: open bassinet in the NBN (E1), working isolet located in the NBN (E2), nonfunctioning isolet in the NBN (E3), nonfunctioning isolet in a quiet room off the NBN (E4), and open bassinet in a quiet room (E5). The number of high noise samples (HNS), the test duration (in seconds), the signal-to-noise ratio (SNR; in dB) measured at bandwidths centered at 1.6, 2.4, 3.2, and 4.0 kHz, and the percentage of neonates with a fail screening outcome based on a common pass-fail screening criteria were compared in the 5 test environments. RESULTS: There were statistically significant differences in the number of HNS accumulated in the 5 test environments (F = 6.79). The use of a nonfunctioning isolet in both the NBN and within a room off the NBN (E3 and E4, respectively) resulted in significantly fewer HNS than when TEOAEs were recorded in the other 3 test environments (E1, E2, and E5). Mean test duration was significantly different among the 5 locations (F = 6. 53). Posthoc analyses revealed that test time in E3 and E4 was significantly shorter than in E1 and E2. The percentage of newborns with a fail (/=2 visits and >/=1 visit during the preceding 18 months) were included in analyses. Immunization coverage levels were calculated and logistic regression was used to estimate the risk of underimmunization associated with different practice and child characteristics. RESULTS: Of the 72 eligible practices that were contacted, 45 participated in the study, yielding a participation rate of 62%. The median immunization coverage of participating offices was 54% (range: 0%-91%). Multivariate analysis revealed 5 independent risk factors for underimmunization. The strongest predictors were having fewer than 50% active children in the practice and children having fewer than 8 visits to the provider. Other significant predictors were the percentage of patients in the practice on Medicaid, administering diphtheria-tetanus pertussis 4 at a separate visit from the Haemophilus influenzae type b booster, and practice location. CONCLUSIONS: These data provide new insights into immunization practices in an important clinical setting that has been poorly characterized previously. Immunization coverage levels were found to be low and significant risk factors for underimmunization were identified. Recommendations are made for immunization policy changes and targeting of immunization improvement interventions at practices that may be at risk for low immunization coverage. immunization, vaccination, immunization programs, primary prevention, private practice, child, preschool, pediatrics, family practice. PMID- 10835087 TI - A proposed solution for addressing the challenge of patient cries for help through an analysis of unsolicited electronic mail. AB - BACKGROUND: Unsolicited electronic mail (e-mail) is e-mail sent to a physician from a person unknown to the physician, who is seeking professional help. The purpose of this project was to analyze unsolicited e-mails sent to a digital textbook author to: 1) characterize the e-mails, 2) determine what resources would be necessary to answer the e-mails, and 3) propose a standard approach to reply to e-mails in a helpful yet medicolegally-responsive manner. MATERIALS AND METHODS: All e-mails (315) sent to a digital textbook author from October 1995 through October 1998 were abstracted. Variables included: date and location, sender type, patient age, subject, medical content, and resources necessary to answer the question. Data frequencies were obtained. RESULTS: The most common location was the.com domain (47.6%). The most common senders were laypersons (66%). Overall, 44.4% of the e-mails concerned children. Detailed, patient specific information was sent in 63.2% of the e-mails. The most common subjects were overviews of a disease or problem (32.4%), differential diagnosis (16.8%), and therapy/treatment questions (15.9%). The medical content covered a broad range of specialties. Specialists were overwhelmingly the resource necessary to answer the e-mails (74.9%). CONCLUSIONS: Pediatricians with educational information on the Internet can expect an increase in the number of unsolicited e mails as Internet usage expands. Laypersons regard even short passages to mean the author is an expert in that particular area. Pediatricians need to consider the ethical and medicolegal implications of responding to unsolicited e-mails. A nonpersonalized, standard e-mail reply is proposed that directs the sender to quality information resources that may be of further assistance. unsolicited electronic mail, e-mail, medical informatics, legal issues, ethical issues, digital libraries. PMID- 10835088 TI - Child care practices in nonindustrialized societies. AB - OBJECTIVE: To described child care practices associated with an increased risk of sudden infant death syndrome (SIDS) in nonindustrialized 19th and 20th century societies. METHODOLOGY: The Human Relations Area Files collection is a unique source of information on the cultures of the world. The electronic version of the Human Relations Area Files represents 200 000 pages of fully indexed cultural material on a stratified random sample of 60 nonindustrialized societies. Using various keyword combinations, texts containing information related to sleeping conditions, social interaction, temperature regulation, feeding, and smoking were identified and using structured questionnaire practices were recorded as being present, absent, or not stated. RESULTS: Relevant information was identified for 53 societies. None of 4 societies mentioning sleep position used the prone position. Swaddling and restraint were commonly practiced, often for extended periods of time. Most information was available on infant feeding and the predominant pattern was of demand breastfeeding for long periods. In many of the cultures, infants are seen as being the focus of attention, in close contact with and under the constant supervision of the mother, siblings, and other relatives. CONCLUSIONS: The study produced no clear evidence that these primarily nonindustrialized societies followed child care practices likely to protect against SIDS. Despite limitations, the information presented should be of interest to those involved in developing reduce the risks programs for SIDS prevention, because it highlights the considerable variation in child care practices and emphasizes that health education messages may not always be cross culturally valid. PMID- 10835089 TI - Health of children adopted from China. AB - OBJECTIVE: Since 1989, American parents have adopted 18 846 Chinese children. This study assesses the health and developmental status of these children after their arrival in the United States. PATIENTS AND METHODS: A total of 452 children (443 girls) in 2 groups were evaluated. The clinic group children (n = 192) included all Chinese adoptees seen in an international adoption clinic between 1991 and 1998. The travel group comprised 260 of 325 Chinese children placed by a single Massachusetts adoption agency between 1991 and 1996 whose adoptive parents and American physicians responded to mailed questionnaires. One hundred ninety one of the travel group children were cared for by 1 of us (N.W.H.) during the adoption process in China. RESULTS: Growth and developmental delays were frequent in the clinic group. Z scores /=2 log(10) VL decline after GT insertion (median: 3.2 log(10)), compared with 7 patients (42%) who had /=2 log(10) VL decline, therapy was changed at the time of or soon after GT insertion (median:.8 months; range: 0-6 months), compared with 7 patients with <2 log(10) VL decline who had therapy changed before GT insertion (median: 3.2 months; range: 1-8 months). Parental questionnaires reported significantly shorter medication administration times after GT insertion, with 70% of patients taking >5 minutes before GT, compared with 0% after GT. Questionnaires indicated satisfaction with GT, with perceived benefits being reduced medication administration time and improved behavior surrounding taking medications. CONCLUSIONS: GT is well-tolerated in pediatric HIV-infected patients and should be considered for selected patients to overcome difficulties with medication administration and to improve adherence. For maximal virologic response, combination therapy should be changed at the time of GT insertion. PMID- 10835094 TI - Prolonged hepatitis B surface antigenemia after vaccination. AB - Infection with hepatitis B virus can lead to serious long-term complications including chronic hepatitis B virus infection leading to hepatocellular carcinoma, liver failure, and death. We report a case of prolonged hepatitis B antigenemia after routine vaccination with Engerix B. A positive hepatitis B surface antigen was found when the individual donated blood 18 days after vaccination. This resulted in rejection of the donated blood and permanent deferral from further donation. It also led to referral to a physician, creating anxiety in the individual and additional unnecessary testing. Additional studies are needed to identify the length to time of hepatitis B surface antigenemia after hepatitis B vaccination, and blood collection centers should be aware of the potential for donors to have a prolonged false-positive hepatitis B surface antigen after vaccination against hepatitis B. hepatitis B, hepatitis B vaccine, hepatitis B surface antigen, vaccine-induced positive hepatitis B surface antigen, Engerix B. PMID- 10835095 TI - Pediatrician and family physician prescription of selective serotonin reuptake inhibitors. AB - OBJECTIVES: Selective serotonin reuptake inhibitor (SSRI) prescriptions for children and adolescents have increased greatly in recent years despite a paucity of demonstrated safety and efficacy data and a lack of clear guidelines for use. Our study sought to describe family physician and pediatrician SSRI prescribing patterns for children and adolescents, identify influences on SSRI prescription variations, and describe the use of SSRI within the overall management of depression and other mental disorders in primary care. DESIGN: A survey was mailed to all 596 active North Carolina general pediatricians and a random sample of 557 family physicians in primary care practice. Family physicians who did not see children in their practice were excluded. The survey instrument consisted of a 4-page questionnaire. Survey items included physician demographics, practice characteristics, general management, volume of pediatric patients with depressive symptoms, prescription of SSRIs for depression and other diagnoses, and potential influences on SSRI prescribing practices. The main outcomes were self-reported physician prescription of SSRIs for children and adolescents. Results were analyzed using chi(2) comparisons and logistic regression. RESULTS: The overall response rate was 66% (55% family physicians and 76% pediatricians). Of the physicians, 72% had prescribed an SSRI for a child or adolescent. Depression was the most common reason for prescribing an SSRI; over two thirds of respondents had prescribed an SSRI for depression in a child 18 years of age or younger. Over half of the physicians reported they had prescribed an SSRI for a diagnosis other than depression in a child 18 years of age or younger. Attention deficit/hyperactivity disorder was the most frequent use cited other than depression, followed by obsessive-compulsive disorder, aggression, eating disorders, and enuresis. Primary care physicians prescribed SSRIs for adolescents more commonly than for younger children. Only 6% of the respondents had ever prescribed an SSRI for a child younger than 6 years of age. In terms of SSRI prescriptions written for depression in the last 6 months, 32% of the physicians had recently prescribed SSRIs for adolescent patients and 6% for patients younger than 12 years of age. Family physicians were more likely than pediatricians to have recently prescribed SSRIs for adolescent patients (41% vs 26%), but there was no difference in recent SSRI prescriptions for children <12 years of age by physician specialty (4% vs 6%). Prescription of SSRIs was not associated with decreased use of counseling for treatment of depression, but prescription of SSRIs was associated with decreased use of referrals (63% vs 74%). There was no difference in the use of counseling between family physicians and pediatricians (61% vs 59%). However, pediatricians were more likely to use referrals in their usual approach to depression (77% vs 48%) compared with family physicians. More family physicians had prescribed SSRIs for pediatric patients compared with pediatricians (91% vs 58%), and more family physicians had prescribed SSRIs in combination with other psychotropic medications (54% vs 31%). For the majority of respondents, SSRI prescriptions constituted most of the medications used to treat childhood depression (75% of family physicians vs 61% of pediatricians). Family physicians were more likely to report a belief in the safety (63% vs 48%) and effectiveness (40% vs 32%) of SSRIs. Only 8% of physicians reported adequate training in the treatment of childhood depression and just 16% were comfortable with the treatment of depression. There were no specialty differences in training for the treatment of childhood depression; however, more family physicians than pediatricians agreed that they were comfortable with the management of childhood depression (22% vs 11%). (ABSTRACT TRUNCATED) PMID- 10835096 TI - Determinants of adolescent physical activity and inactivity patterns. AB - OBJECTIVES: Despite recognition of the important influence of environmental determinants on physical activity patterns, minimal empirical research has been done to assess the impact of environmental/contextual determinants of physical activity. This article aims to investigate environmental and sociodemographic determinants of physical activity and inactivity patterns among subpopulations of US adolescents. We define environmental determinants as modifiable factors in the physical environment that impose a direct influence on the opportunity to engage in physical activity. The present research examines environmental and sociodemographic determinants of physical activity and inactivity with the implication that these findings can point toward societal-level intervention strategies for increasing physical activity and decreasing inactivity among adolescents. STUDY DESIGN AND METHODOLOGY: The study population consists of nationally representative data from the 1996 National Longitudinal Study of Adolescent Health on 17 766 US adolescents enrolled in US middle and high schools (including 3933 non-Hispanic blacks, 3148 Hispanics, and 1337 Asians). Hours/week of inactivity (TV/video viewing and video/computer games) and times/week of moderate to vigorous physical activity were collected by questionnaire. Outcome variables were moderate to vigorous physical activity and inactivity, which were broken into categories (physical activity: 0-2 times/week, 3-4 times/week, and >/=5 times/week; inactivity: 0-10 hours/week, 11-24 hours/week, and >/=25 hours/week). Sociodemographic and environmental correlates of physical activity and inactivity were used as exposure and control variables and included sex, age, urban residence, participation in school physical education program, use of community recreation center, total reported incidents of serious crime in neighborhood, socioeconomic status, ethnicity, generation of residence in the United States, presence of mother/father in household, pregnancy status, work status, in-school status, region, and month of interview. Logistic regression models of high versus low and medium physical activity and inactivity were used to investigate sex and ethnic interactions in relation to environmental and sociodemographic factors to examine evidence for the potential impact of physical education and recreation programs and sociodemographic factors on physical activity and inactivity patterns. RESULTS: Moderate to vigorous physical activity was lower and inactivity higher for non-Hispanic black and Hispanic adolescents. Participation in school physical education programs was considerably low for these adolescents and decreased with age. Participation in daily school physical education (PE) program classes (adjusted odds ratio [AOR]: 2.21; confidence interval [CI]: 1.82-2.68) and use of a community recreation center (AOR: 1.75; CI: 1.56-1.96) were associated with an increased likelihood of engaging in high level moderate to vigorous physical activity. Maternal education was inversely associated with high inactivity patterns; for example, having a mother with a graduate or professional degree was associated with an AOR of.61 (CI:.48-.76) for high inactivity. High family income was associated with increased moderate to vigorous physical activity (AOR: 1.43; CI: 1.22-1.67) and decreased inactivity (AOR:.70; CI:.59-.82). High neighborhood serious crime level was associated with a decreased likelihood of falling in the highest category of moderate to vigorous physical activity (AOR:.77; CI:.66-.91). CONCLUSIONS: These results show important associations between modifiable environmental factors, such as participation in school PE and community recreation programs, with activity patterns of adolescents. Despite the marked and significant impact of participation in school PE programs on physical activity patterns of US adolescents, few adolescents participated in such school PE programs; only 21.3% of all adolescents PMID- 10835097 TI - Infant massage compared with crib vibrator in the treatment of colicky infants. AB - OBJECTIVE: To evaluate the effectiveness of infant massage compared with that of a crib vibrator in the treatment of infantile colic. METHODS: Infants <7 weeks of age and perceived as colicky by their parents were randomly assigned to an infant massage group (n = 28) or a crib vibrator group (n = 30). Three daily intervention periods were recommended in both groups. Parents recorded infant crying and given interventions in a structured cry diary that was kept for 1 week before (baseline) and for 3 weeks during the intervention. Parents were interviewed after the first and third weeks of intervention to obtain their evaluation of the effectiveness of the given massage or crib vibration. RESULTS: At baseline, the mean amount of total crying was 3.6 (standard deviation: 1.4) hours/day in the massage group infants and 4.2 (2.0) hours/day in the vibrator group infants. The mean amount of colicky crying was 2.1 (standard deviation: 1.1) hours/day and 2.9 (1.5) hours/day, respectively. The mean number of daily intervention periods was 2.2 in both groups. Over the 4-week study, the amount of total and colicky crying decreased significantly in both intervention groups. The reduction in crying was similar in the study groups: total crying decreased by a mean 48% in the massage group and by 47% in the vibrator group, and colicky crying decreased by 64% and 52%, respectively. The amount of other crying (total crying minus colicky crying) remained stable in both groups over the intervention. Ninety-three percent of the parents in both groups reported that colic symptoms decreased over the 3-week intervention, and 61% of the parents in the massage group and 63% of the parents in the crib vibrator group perceived the 3-week intervention as colic reducing. CONCLUSIONS: Infant massage was comparable to the use of a crib vibrator in reducing crying in colicky infants. We suggest that the decrease of total and colicky crying in the present study reflects more the natural course of early infant crying and colic than a specific effect of the interventions. PMID- 10835098 TI - Georgia's breastfeeding promotion program for low-income women. AB - OBJECTIVE: Beginning in 1990, Georgia's Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) initiated 5 new strategies to promote breastfeeding among its pregnant and postpartum clients. These strategies were implemented in 1991, each to be provided as an addition to its standard program of counseling on breastfeeding and distributing appropriate literature: 1) enhanced breastfeeding education, 2) breast pump loans, 3) hospital-based programs, 4) peer counseling, and 5) community coalitions. The enhanced breastfeeding education strategy provides access to a hotline as well as periodic training of staff, and the breast pump loan provides free breast pumps to mothers who want to use them. The hospital-based strategy provides bedside support and counseling to women who have just given birth and includes staff training, as well as a hotline number for women to call after they leave the hospital. The peer-counseling strategy focuses on identifying former WIC participants who have successfully breastfed their infants; these women are recruited to provide support and encouragement to current WIC participants. Finally, the community coalitions approach is designed to identify existing community attitudes about breastfeeding, establish plans to address gaps in breastfeeding services, to develop resource guides on breastfeeding for the community, and to advocate at the community level to support breastfeeding women. The objective of our research was to evaluate the impact of breastfeeding promotion strategies on breastfeeding initiation among WIC participants in Georgia. METHODS: Using data from the Pregnancy Nutrition Surveillance System (PNSS) for 1992-1996, we examined breastfeeding initiation rate during this period and compared rates among 6 different intervention strategies. Also, we used data from the Pregnancy Risk Assessment Monitoring System (PRAMS) to assess breastfeeding initiation and duration among WIC enrollees. We conducted 13 focus groups to understand the experiences of program participants. Ten focus groups were conducted with women who were breastfeeding their infants, 3 each with women from the community coalitions, hospital-based programs, and standard education programs, and 1 with women from the breast pump loan program. Three focus groups were conducted with women who were feeding their infants formula. RESULTS: PNSS data show that breastfeeding initiation increased in the Georgia WIC program from 31.6% in 1992 to 39.5% in 1996. PRAMS data confirmed the increase in breastfeeding initiation from 33.6% (standard error [SE]: 2.2) in 1993 to 42.1% (SE: 2.4) in 1996 among WIC participants. Both datasets (PRAMS and PNSS) showed breastfeeding initiation to be well below the year 2000 goal of 75%. Overall, PRAMS data show a high breastfeeding initiation among non-WIC participants (range: 64.7% [SE: 2.2]) for 1994 to 70.1% (SE: 2.2) in 1996. The percent change between 1993 and 1996 was 8% for non-WIC participants, and it was 25% for the WIC participants among those responding to the PRAMS questionnaire. Data from PRAMS indicated no statistical change in the percentage of WIC enrollees who breastfed their infants for 8 weeks or more; this estimate was 18.3% (95% confidence interval (CI): 14.9-21.8) in 1993 and 19.4% (95% CI: 15.7-23.2) in 1996, well below the Healthy People 2000 objective of 50% at 6 months. According to PNSS data, the largest increases in breastfeeding initiation for 1992 to 1996 were among younger women ( His and Thr142 --> His, lie, respectively, on the surface of the N- and C-terminal domains on opposite sides of the active site cleft. The design takes advantage of hinge-bending motion which allows the binding site to adapt to the most favorable ligand geometry for the metal. Introduction of the two histidines increases the melting temperature of the protein by 2.0 degrees C at pH 7.4. Metal binding further increases the melting temperature, but only by a small amount (up to 1.5 degrees C). A third substitution, Gln141 --> His, which could act as a third ligand in principle, does not do so, demonstrating the difficulty in mimicking naturally occurring metal-binding sites. PMID- 10835105 TI - Improvement in the efficiency of formyl transfer of a GAR transformylase hybrid enzyme. AB - A hybrid glycinamide ribonucleotide transformylase was assembled from two protein domains that were treated as discrete modules. One module contained the ribonucleotide binding domain from the purN glycinamide ribonucleotide transformylase; the second module contained the catalytic machinery and the formyl tetrahydrofolate binding domain from the enzyme encoded by purU, formyl tetrahydrofolate hydrolase. The resultant enzyme showed 0.1% catalytic activity of the wild-type glycinamide ribonucleotide transformylase enzyme but had a formyl transfer efficiency of 10%. A combinatorial mutagenesis approach was used to improve the solubility and formyl transfer properties of the hybrid enzyme. The mutagenized hybrid glycinamide ribonucleotide transformylase was initially expressed as a fusion to the alpha-peptide of beta-galactosidase. Clones were selected for improvement in solubility by determining which clones were capable of alpha-complementation using a blue/white screen. One clone was further characterized and found to have an improved efficiency of transfer of the ribonucleotide increasing this property to >95%. PMID- 10835106 TI - Linker insertion mutagenesis based on IS21 transposition: isolation of an AMP insensitive variant of catabolic ornithine carbamoyltransferase from Pseudomonas aeruginosa. AB - The bacterial insertion sequence IS21 when repeated in tandem efficiently promotes non-replicative cointegrate formation in Escherichia coli. An IS21-IS21 junction region which had been engineered to contain unique SalI and BglII sites close to the IS21 termini was not affected in the ability to form cointegrates with target plasmids. Based on this finding, a novel procedure of random linker insertion mutagenesis was devised. Suicide plasmids containing the engineered junction region (pME5 and pME6) formed cointegrates with target plasmids in an E.coli host strain expressing the IS21 transposition proteins in trans. Cointegrates were resolved in vitro by restriction with SalI or BglII and ligation; thus, insertions of four or 11 codons, respectively, were created in the target DNA, practically at random. The cloned Pseudomonas aeruginosa arcB gene encoding catabolic ornithine carbamoyltransferase was used as a target. Of 20 different four-codon insertions in arcB, 11 inactivated the enzyme. Among the remaining nine insertion mutants which retained enzyme activity, three enzyme variants had reduced affinity for the substrate ornithine and one had lost recognition of the allosteric activator AMP. The linker insertions obtained illustrate the usefulness of the method in the analysis of structure-function relationships of proteins. PMID- 10835107 TI - Study of B72.3 combining sites by molecular modeling and site-directed mutagenesis. AB - A B72.3 Fab/sTn(2) complex was modeled from the known structure of B72.3 Fab and the dimeric Tn-serine cluster (sTn(2)). In the complex model, the side chains of 15 heavy- and light-chain complementarity-determining region (CDR) residues and the main chains of two light-chain CDR residues contact the sTn(2) epitope. Among 15 CDR residues which contact sTn(2) in the model, two heavy-chain residues (Ser95 and Tyr97) and light-chain CDR residue (Tyr96) have been confirmed in a previous study. To test the accuracy of the computational model, further site directed mutagenesis was performed by alanine scanning on the remaining 12 residues that are predicted in the model to have side-chain interactions with sTn(2). Of these 12 mutants, eight that are all from the heavy-chain (His32Ala, Ala33Leu, Tyr50Ala, Ser52Ala, Asn52Ala, Asp56Ala, Lys58Ala and Tyr96Ala) had significantly reduced sTn(2) affinities, and four consisting of three light-chain mutations (Asn32Ala, Trp92Ala and Thr94Ala) and one heavy-chain mutation (His35Ala) retained wild-type sTn(2) affinity. On the whole, this evidence suggests that the complex model, although not perfect, is correct in many of its features. In a more general vein, these results lend credibility to the computational modeling approach for the study of the molecular basis of antigen antibody complexes. PMID- 10835108 TI - Structure of the Malpha2-3 toxin alpha antibody-antigen complex: combination of modelling with functional mapping experimental results. AB - Modelled structures of the acetylcholine receptor-mimicking antibody, Malpha2-3, both free and bound to its antigen, toxin alpha, are assessed in the light of new experimental mutational data from functional mapping of the paratopic region of Malpha2-3. The experimental results are consistent with the previously-predicted structure of the free antibody, and also demonstrate that structural particularities of the Malpha2-3 combining site that were identified in the models play a role in the protein association. The modelled conformations of the hypervariable loops are discussed in the context of recent new data and analyses. The new mutational data allow several previously-considered modelled structures of the complex to be rejected. Two quite similar models now remain. PMID- 10835109 TI - Design and synthesis of germline-based hemi-humanized single-chain Fv against the CD18 surface antigen. AB - The 6.7 murine monoclonal antibody (mAb) recognizes the human CD18 antigen and is therefore of interest as an anti-inflammatory agent. The 6.7 heavy variable chain (VH) was humanized using the closest human germline sequence as the template on to which to graft the murine complementary determining regions (CDRs). Two versions were proposed, one in which the residue proline 45 of the murine form was maintained and another in which this framework residue was changed to the leucine found in the human sequence. These VH humanized versions were expressed in the yeast Pichia pastoris as hemi-humanized single-chain Fv (scFvs), with the VL from the murine antibody. The scFv from the murine antibody was also expressed. The binding activities of the murine and both hemi-humanized scFvs were determined by flow cytometry analysis. All the constructions were able to recognize human lymphocytes harboring CD18, indicating successful humanization with transfer of the original binding capability. Some differences between the two hemi-humanized versions were observed. The method used was simple and straightforward, with no need for refined structural analyses and could be used for the humanization of other antibodies. PMID- 10835110 TI - An efficient route to the production of an IgG-like bispecific antibody. AB - Production of IgG-form bispecific antibody (BsAb-IgG) by co-expressing two antibodies in transfected cells is often inefficient owing to the unwanted pairing between the component heavy and light chains. We have developed an efficient method for the production of a novel IgG-like BsAb by using the natural dimerization mechanism between IgG heavy and light chains. Two single-chain Fv (scFv) of different specificity are fused to the constant domain of human kappa chain (C(L)) and the first constant domain of human heavy chain (C(H1)), to form two polypeptides, (scFv)(1)-C(L) and (scFv)(2)-C(H1)-C(H2)-C(H3), respectively. Co-expression of the two polypeptides in mammalian cells results in the formation of a covalently linked IgG-like hetero-tetramer, Bs(scFv)(4)-IgG, with dual specificity. Our approach yields a homogeneous bispecific IgG-like antibody product with each molecule containing four antigen binding sites, two for each of its target antigens. A Bs(scFv)(4)-IgG was prepared using two scFv antibodies each directed against a different epitope of a vascular endothelial growth factor receptor, the kinase insert domain-containing receptor (KDR). The Bs(scFv)(4)-IgG is capable of simultaneously binding to the two epitopes on the receptor. Further, the Bs(scFv)(4)-IgG also retains the antigen-binding efficacy and biological activity of its component antibodies. PMID- 10835111 TI - Fluorolabeling of antibody variable domains with green fluorescent protein variants: application to an energy transfer-based homogeneous immunoassay. AB - A site-specific and efficient fluorolabeling of antibody variable regions with green fluorescent protein (GFP) variants and its application to an energy transfer-based homogeneous fluoroimmunoassay (open sandwich FIA) were attempted. Two chimeric proteins, Trx-V(H)-EBFP and Trx-V(L)-EGFP, consisting of V(H) and V(L) fragments of anti-hen egg lysozyme (HEL) antibody HyHEL-10 and two GFP color variants, EBFP and EGFP, respectively, were designed to be expressed in cytoplasm of trxB - mutant Escherichia coli as fusions with thioredoxin from E.coli The mixture of two proteins could be purified with HEL-affinity chromatography, retaining sufficient intrinsic fluorescence and binding activity to HEL. A significant increase in fluorescence resonance energy transfer (FRET) dependent on HEL concentration was observed, indicating the reassociation of the V(H) and V(L) domains of these chimeric proteins due to co-existing antigen. With this open sandwich FIA, an HEL concentration of 1-100 microg/ml could be non competitively determined. The assay could be performed in a microplate format and took only a few minutes to obtain a sufficient signal after simple mixing of the chimeric proteins with samples. This represents the first demonstration that the FRET between GFP variants is applicable to homogeneous immunoassay. PMID- 10835112 TI - Functional expression of horseradish peroxidase in Saccharomyces cerevisiae and Pichia pastoris. AB - The ability to engineer proteins by directed evolution requires functional expression of the target polypeptide in a recombinant host suitable for construction and screening libraries of enzyme variants. Bacteria and yeast are preferred, but eukaryotic proteins often fail to express in active form in these cells. We have attempted to resolve this problem by identifying mutations in the target gene that facilitate its functional expression in a given recombinant host. Here we examined expression of HRP in Saccharomyces cerevisiae. Through three rounds of directed evolution by random point mutagenesis and screening, we obtained a 40-fold increase in total HRP activity in the S.cerevisiae culture supernatant compared with wild-type, as measured on ABTS ?2, 2'-azinobis(3 ethylbenzthiazoline-6-sulfonic acid) (260 units/l/OD(600)). Genes from wild-type and two high-activity clones were expressed in Pichia pastoris, where the total ABTS activity reached 600 units/l/OD(600) in shake flasks. The mutants show up to 5.4-fold higher specific activity towards ABTS and 2.3-fold higher specific activity towards guaiacol. PMID- 10835114 TI - Delmar J. Stauffer, RSNA executive director: A tribute PMID- 10835113 TI - CT evaluation of prosthetic ossicular reconstruction procedures: what the otologist needs to know. AB - Postoperative otologic evaluation of patients who have undergone ossicular reconstruction is often difficult. However, thin-section computed tomography (CT) can help determine the type of prosthesis used for reconstruction and adequately assess for complications that may be causing postoperative conductive hearing loss. A variety of prostheses may be used in ossicular reconstruction (eg, stapes prosthesis, incus interposition graft, Applebaum prosthesis, Black oval-top prosthesis, Richards centered prosthesis, Goldenberg prosthesis) and can usually be identified at CT by their shapes and locations. Several causes of prosthetic failure are readily demonstrated at CT, including recurrent cholesteatoma and otitis media, formation of granulation tissue or adhesions, and various mechanical problems (eg, subluxation, dislocation, extrusion, fracture, bending). Perilymphatic fistula can be difficult to identify at CT but may be suggested by the presence of pneumolabyrinth, unexplained middle ear effusion, or fluid accumulation within the mastoid air cells. The presence of soft tissue within the oval window niche 4-6 weeks following surgery may indicate poststapedectomy granuloma or fibrosis. Familiarity with the normal and abnormal CT appearances of ossicular prostheses will enable the radiologist to assist the otologist in identifying patients in whom revision surgery is most appropriate. PMID- 10835115 TI - Complications of renal transplantation: evaluation with US and radionuclide imaging. AB - Following renal transplantation, patients are often evaluated with ultrasonography (US) or radionuclide imaging to assess renal function and the presence of possible complications. Both modalities are inexpensive, noninvasive, and nonnephrotoxic. A basic understanding of the surgical techniques commonly used for renal transplantation is useful when imaging these patients in order to recognize complications and to direct further imaging or intervention. The most frequent complications of renal transplantation include perinephric fluid collections; decreased renal function; and abnormalities of the vasculature, collecting system, and renal parenchyma. Perinephric fluid collections are common following transplantation, and their clinical significance depends on the type, location, size, and growth of the fluid collection, features that are well evaluated with US. Causes of diminished renal function include acute tubular necrosis, rejection, and toxicity from medications. Radionuclide imaging is the most useful modality for assessing renal function. Vascular complications of transplantation include occlusion or stenosis of the arterial or venous supply, arteriovenous fistulas, and pseudoaneurysms. Although the standard for evaluating these vascular complications is angiography, US is an excellent noninvasive method for screening. Other transplant complications such as abnormalities of the collecting system and renal parenchyma are well-evaluated with both radionuclide imaging and US. PMID- 10835116 TI - MR imaging classification of perianal fistulas and its implications for patient management. AB - Until recently, imaging had a limited role in the preoperative assessment of perianal fistulas. Magnetic resonance (MR) imaging has been shown to demonstrate accurately the anatomy of the perianal region. In addition to showing the anal sphincter mechanism, MR imaging clearly shows the relationship of fistulas to the pelvic diaphragm (levator plate) and the ischiorectal fossae. This relationship has important implications for surgical management and outcome and has been classified into five MR imaging-based grades. If the ischioanal and ischiorectal fossae are unaffected, disease is likely confined to the sphincter complex (simple intersphincteric fistulization, grade 1 or 2), and outcome following simple surgical management is favorable. Involvement of the ischioanal or ischiorectal fossa by a fistulous track or abscess indicates complex disease related to trans-sphincteric or suprasphincteric disease (grade 3 or 4). Correspondingly more complex surgery may be required that may threaten continence or may require colostomy to allow healing. If the track traverses the levator plate, a translevator fistula (grade 5) is present, and a source of pelvic sepsis should be sought. PMID- 10835118 TI - Spectrum of congenital anomalies of the inferior vena cava: cross-sectional imaging findings. AB - Congenital anomalies of the inferior vena cava (IVC) and its tributaries have become more commonly recognized in asymptomatic patients. The embryogenesis of the IVC is a complex process involving the formation of several anastomoses between three paired embryonic veins. The result is numerous variations in the basic venous plan of the abdomen and pelvis. A left IVC typically ends at the left renal vein, which crosses anterior to the aorta to form a normal right-sided prerenal IVC. In double IVC, the left IVC typically ends at the left renal vein, which crosses anterior to the aorta to join the right IVC. In azygos continuation of the IVC, the prerenal IVC passes posterior to the diaphragmatic crura to enter the thorax as the azygos vein. In circumaortic left renal vein, one left renal vein crosses anterior to the aorta and another crosses posterior to the aorta. In retroaortic left renal vein, the left renal vein passes posterior to the aorta. In circumcaval ureter, the proximal ureter courses posterior to the IVC. Other anomalies include absence of the infrarenal IVC or the entire IVC. These anomalies can have significant clinical implications. Awareness of these anomalies is necessary to avoid diagnostic pitfalls. PMID- 10835119 TI - US in the diagnosis of pediatric chest diseases. AB - Most pediatric chest diseases are adequately evaluated with chest radiography. However, when chest radiography does not allow identification of the location and nature of an area of increased opacity, ultrasonography (US) can help establish the diagnosis. US may be helpful in evaluation of persistent or unusual areas of increased opacity in the peripheral lung, pleural abnormalities, and mediastinal widening; US is particularly useful in patients with complete opacification of a hemithorax at radiography. In cases of pulmonary parenchymal lesions, identification of air or fluid bronchograms at US and of pulmonary vessels at color flow imaging is useful for differentiating pulmonary consolidation or atelectasis from lung masses and pleural lesions. US allows characterization of pleural fluid collections as simple, complicated, or fibroadhesive, which is important information for planning thoracentesis or thoracotomy. Computed tomography and magnetic resonance imaging are superior to US in evaluation of the mediastinum, but US is a reasonable alternative in certain situations (eg, to avoid unnecessary investigation of a normal thymus simulating a mediastinal mass). In cases of chest wall lesions, US may enable localization of the site of origin to soft tissues or an extrapleural intrathoracic location. Osseous involvement, particularly rib involvement, is easily evaluated with US. PMID- 10835121 TI - Practice corner: doing business with the devil PMID- 10835120 TI - Aspiration diseases: findings, pitfalls, and differential diagnosis. AB - The aspiration of different substances into the airways and lungs may cause a variety of pulmonary complications. These disease entities most commonly involve the posterior segment of the upper lobes and the superior segment of the lower lobes. Esophagography and computed tomography (CT) are especially useful in the evaluation of aspiration disease related to tracheoesophageal or tracheopulmonary fistula. Foreign body aspiration typically occurs in children and manifests as obstructive lobar or segmental overinflation or atelectasis. An extensive, patchy bronchopneumonic pattern may be observed in patients following massive aspiration of gastric acid or water. CT is the modality of choice in establishing the diagnosis of exogenous lipoid pneumonia, which can result from aspiration of hydrocarbons or of mineral oil or a related substance. Aspiration of infectious material manifests as necrotizing consolidation and abscess formation. The relatively low diagnostic accuracy of chest radiography in aspiration diseases can be improved with CT and by being familiar with the clinical settings in which specific complications are likely to occur. Recognition of the varied clinical and radiologic manifestations of these disease entities is imperative for prompt, accurate diagnosis, resulting in decreased morbidity and mortality rates. PMID- 10835122 TI - Diseases of the hepatopulmonary axis. AB - Hepatopulmonary syndrome is the most widely recognized of the processes associated with end-stage liver disease. Chronic liver dysfunction is associated with pulmonary manifestations due to alterations in the production or clearance of circulating cytokines and other mediators. Hepatopulmonary syndrome results in hypoxemia due to pulmonary vasodilatation with significant arteriovenous shunting and ventilation-perfusion mismatch. Hepatic hydrothorax may develop in patients with cirrhosis and ascites. Rarely, pulmonary hypertension occurs in the setting of portal hypertension. A second group of disorders may primarily affect the lungs and liver (the hepatopulmonary axis). Among these are the congenital conditions alpha(1)-antitrypsin deficiency and cystic fibrosis. Autoimmune liver disease may be associated with lymphocytic interstitial pneumonitis, fibrosing alveolitis, intrapulmonary granulomas, and bronchiolitis obliterans with organizing pneumonia. Sarcoidosis affects the lung and liver in up to 70% of patients. Medications such as amiodarone can result in a characteristic radiologic appearance of pulmonary and hepatic toxic effects. Knowledge of these associations will assist the radiologist in forming a meaningful differential diagnosis and may influence treatment decisions. PMID- 10835123 TI - Metal artifact reduction sequence: early clinical applications. AB - Artifact arising from metal hardware remains a significant problem in orthopedic magnetic resonance imaging. The metal artifact reduction sequence (MARS) reduces the size and intensity of susceptibility artifacts from magnetic field distortion. The sequence, which is based on view angle tilting in combination with increased gradient strength, can be conveniently used in conjunction with any spin-echo sequence and requires no additional imaging time. In patients with persistent pain after femoral neck fracture, the MARS technique allows visualization of marrow adjacent to hip screws, thus enabling diagnosis or exclusion of avascular necrosis. Other applications in the hip include assessment of periprosthetic soft tissues after hip joint replacement surgery, postoperative assessment after resection of bone tumors and reconstruction, and localization of unopacified methyl methacrylate cement prior to hip arthroplasty revision surgery. In the knee, the MARS technique allows visualization of structures adjacent to implanted metal staples, pins, or screws. The technique can significantly improve visualization of periprosthetic bone and soft-tissue structures even in patients who have undergone total knee arthroplasty. In patients with spinal fixation hardware, the MARS technique frequently allows visualization of the vertebral bodies and spinal canal contents. The technique can be helpful after wrist fusion or screw fixation of scaphoid fractures. PMID- 10835124 TI - Imaging features of primary and recurrent esophageal cancer at FDG PET. AB - Because of the poor prognosis for patients with esophageal cancer and the risks associated with surgical intervention, accurate staging is essential for optimal treatment planning. Positron emission tomography (PET) with 2-[fluorine-18]fluoro 2-deoxy-d-glucose (FDG) is a useful adjunct to more conventional imaging modalities in this setting. FDG PET is not an appropriate first-line diagnostic procedure in the detection of esophageal cancer and is not helpful in detecting local invasion by the primary tumor, and further studies are required to determine its efficacy in the detection of local nodal metastases. However, FDG PET is superior to anatomic imaging modalities in the ability to detect distant metastases. Metastases to the liver, lungs, and skeleton can readily be identified at FDG PET. In addition, FDG PET has proved valuable in determining the resectability of disease and allows scanning of a larger volume than is possible with computed tomography. Recurrent disease is readily diagnosed and differentiated from scar tissue with FDG PET. In addition, FDG PET may play a valuable role in the follow-up of patients who undergo chemotherapy and radiation therapy, allowing early changes in treatment for unresponsive tumors. The management of most patients with esophageal cancer can be improved with use of FDG PET. PMID- 10835125 TI - Tailored helical CT evaluation of acute abdomen. AB - Helical computed tomography (CT) allows rapid, cost-effective evaluation of patients with acute abdominal pain. Tailoring the examination to the working clinical diagnosis by optimizing constituent factors (eg, timing of acquisition, contrast material used, means and rate of contrast material administration, collimation, pitch) can markedly improve diagnostic accuracy. Rapid (>/=3 mL/sec) intravenous injection of contrast material is required for optimal assessment of acute pancreatitis, ischemic bowel, aortic aneurysm, and aortic dissection. Narrow collimation and small reconstruction intervals can help detect calculi in the biliary system and genitourinary tract. Tailored helical CT in patients with acute pyelonephritis usually involves several acquisitions through the kidneys during various phases of renal enhancement. In patients with suspected renal infarction, CT protocol must include an acquisition during the corticomedullary phase. Helical CT with 5-mm collimation through the lower abdomen and pelvis is used to evaluate patients with suspected diverticulitis. Use of both oral and intravenous contrast material can help localize small bowel perforation and characterize related complications. Tailored helical CT for assessment of abdominal hemorrhage consists of initial unenhanced CT followed by optional contrast material-enhanced CT. Clear communication between the radiologist, the patient, and the referring physician is essential for narrowing the differential diagnosis into a working diagnosis prior to helical CT. PMID- 10835126 TI - Gallbladder stones: imaging and intervention. AB - Imaging of the gallbladder for cholelithiasis and its complications has changed dramatically in recent decades along with expansion of interventional techniques related to the disease. Ultrasonography (US) is the method of choice for detection of gallstones. The characteristic US findings of gallstones are a highly reflective echo from the anterior surface of the gallstone, mobility of the gallstone on repositioning the patient, and marked posterior acoustic shadowing. Oral cholecystography remains an excellent method of gallstone detection, but its role has been limited due to the advantages of US. Most people with cholelithiasis will not experience symptoms or complications related to gallstones. When biliary colic does occur, it is typically caused by transient obstruction of the cystic duct by a stone. The primary imaging modality in suspected acute calculous cholecystitis is usually US or cholescintigraphy. Detection of gallstones alone does not permit a diagnosis of acute cholecystitis; however, secondary US findings provide more specific information. In detection of choledocholithiasis, endoscopic retrograde cholangiopancreatography and magnetic resonance cholangiopancreatography are superior to US. In certain clinical settings, interventional radiologic procedures have become an important alternative to surgery in the treatment of gallstones and their complications; techniques include percutaneous cholecystostomy and gallstone removal. PMID- 10835127 TI - Hepatobiliary and pancreatic manifestations of cystic fibrosis: MR imaging appearances. AB - Magnetic resonance (MR) imaging and MR cholangiopancreatography are useful, noninvasive techniques for the assessment of pancreatic and hepatobiliary complications in cystic fibrosis. Abnormalities of the pancreas in cystic fibrosis are typically characterized by fat deposition, which has increased signal intensity on T1-weighted MR images, and pancreatic fibrosis, which has low signal intensity on both T1- and T2-weighted images. Pancreatic cysts are a relatively common finding; these cysts are typically quite small but are well demonstrated at MR imaging and MR cholangiopancreatography. Pancreatic duct abnormalities are also occasionally seen. Hepatic manifestations range from hepatomegaly and diffuse fatty infiltration to severe cirrhosis with fibrotic change, regenerative nodules, and portal hypertension. Splenomegaly is often characterized by siderotic nodules that manifest as multiple focal areas of abnormal low signal intensity within the spleen. Biliary manifestations include cholelithiasis, stricturization, and narrowing or dilatation of intra- and extrahepatic bile ducts. Gallbladder abnormalities including microgallbladder are also readily demonstrated. MR cholangiopancreatography can be used to help determine the presence and severity of biliary complications without resorting to more invasive procedures and, in conjunction with MR imaging, may prove useful in the assessment of patients with cystic fibrosis who present with abdominal symptoms that suggest hepatobiliary involvement. PMID- 10835128 TI - Radiologic features of vasculitis involving the gastrointestinal tract. AB - Vasculitides can cause local or diffuse pathologic changes in the gastrointestinal tract, resulting in nonspecific paralytic ileus, mesenteric ischemia, submucosal edema and hemorrhage, or bowel perforation or stricture. The extent and clinical course of disease depend on the size and location of the affected vessel and the histologic characteristics of the lesion. Vasculitis may primarily involve large vessels (eg, giant cell arteritis, Takayasu arteritis), medium-sized vessels (eg, polyarteritis nodosa, Kawasaki disease, primary granulomatous central nervous system vasculitis), or small vessels (eg, Wegener granulomatosis, Churg-Strauss syndrome, microscopic polyangiitis, Henoch Schonlein syndrome, systemic lupus erythematosus, rheumatoid vasculitis, Behcet syndrome). Radiologic findings in various types of vasculitis often overlap considerably and therefore have limited value in making a specific diagnosis. Nevertheless, the possibility of vasculitis should be considered whenever mesenteric ischemic changes occur in young patients, are noted at unusual sites (eg, stomach, duodenum, rectum), have a tendency to concomitantly involve the small and large intestine, and are associated with genitourinary involvement. Knowledge of systemic clinical manifestations in affected patients may suggest and even help establish the specific diagnosis. PMID- 10835129 TI - Hydatid disease: radiologic and pathologic features and complications. AB - Hydatid disease primarily affects the liver and typically demonstrates characteristic imaging findings. However, there are many potential local complications (eg, intrahepatic complications, exophytic growth, transdiaphragmatic thoracic involvement, perforation into hollow viscera, peritoneal seeding, biliary communication, portal vein involvement, abdominal wall invasion). Furthermore, secondary involvement due to hematogenous dissemination may be seen in almost any anatomic location (eg, lung, kidney, spleen, bone, brain). Ultrasonography (US) is particularly useful for the detection of cystic membranes, septa, and hydatid sand. Computed tomography (CT) best demonstrates cyst wall calcification and cyst infection. CT and magnetic resonance (MR) imaging may demonstrate cyst wall defects as well as the passage of contents through a defect. Chest radiography, US, CT, and MR imaging are all useful in depicting transdiaphragmatic migration of hydatid disease. CT is the modality of choice in peritoneal seeding. US and CT demonstrate rupture in most cases that involve wide communication. Indirect signs of biliary communication include increased echogenicity at US and fluid levels and signal intensity changes at MR imaging. CT allows precise assessment of osseous lesions, whereas MR imaging is superior in demonstrating neural involvement. Familiarity with atypical manifestations of hydatid disease may be helpful in making a prompt, accurate diagnosis. PMID- 10835130 TI - Radiologic history exhibit. Musculoskeletal eponyms: who are those guys? PMID- 10835132 TI - Illuminations PMID- 10835131 TI - Autosomal recessive polycystic kidney disease: radiologic-pathologic correlation. AB - Autosomal recessive polycystic kidney disease is a heritable but phenotypically variable disorder characterized by varying degrees of nonobstructive renal collecting duct ectasia, hepatic biliary duct ectasia and malformation, and fibrosis of both liver and kidneys. In the kidney, the dilated collecting ducts and interstitial fibrosis, when severe, may significantly impair renal function and result in hypertension and renal failure. Imaging typically shows large but reniform kidneys, diffusely increased renal parenchymal echogenicity at ultrasonography, and a striated nephrogram after contrast material administration. In the liver, periportal fibrosis accompanies the malformed and dilated bile ducts; this may result in portal hypertension. The liver may appear normal or may show intrahepatic biliary dilatation; once portal hypertension develops, splenomegaly and varices are usually evident. The relative degrees of kidney and liver involvement tend to be inverse: Children with severe renal disease usually have milder hepatic disease, and those with severe hepatic disease tend to evidence mild renal impairment. Presently, treatment consists of supportive management and control of hypertension. Replacement therapy for renal failure (dialysis or kidney transplantation) and control of portal hypertension (portal circulatory diversion or liver transplantation) may be necessary. PMID- 10835133 TI - Elastodynamic shape modeler: a tool for defining the deformation behavior of virtual tissues. AB - A main goal of surgical simulators is the creation of virtual training environments for prospective surgeons. Thus, students can rehearse the various steps of surgical procedures on a computer system without any risk to the patient. One main condition for realistic training is the simulated interaction with virtual medical devices, such as endoscopic instruments. In particular, the virtual deformation and transection of tissues are important. For this application, a neuro-fuzzy model has been developed, which allows the description of the visual and haptic deformation behavior of the simulated tissue by means of expert knowledge in the form of medical terms. Pathologic conditions affecting the visual and haptic tissue response can be easily changed by a medical specialist without mathematical knowledge. By using the personal computer-based program Elastodynamic Shape Modeler, these conditions can be adjusted via a graphical user interface. With a force feedback device, which is similar to a real laparoscopic instrument, virtual deformations can be performed and the resulting haptic feedback can be felt. Thus, use of neuro-fuzzy technologies for the definition and calculation of virtual deformations seems applicable to the simulation of surgical interventions in virtual environments. PMID- 10835134 TI - Interfacing the PACS and the HIS: results of a 5-year implementation. AB - An interface was created between the Department of Defense's hospital information system (HIS) and its two picture archiving and communication system (PACS)-based radiology information systems (RISs). The HIS is called the Composite Healthcare Computer System (CHCS), and the RISs are called the Medical Diagnostic Imaging System (MDIS) and the Digital Imaging Network (DIN)-PACS. Extensive mapping between dissimilar data protocols was required to translate data from the HIS into both RISs. The CHCS uses a Health Level 7 (HL7) protocol, whereas the MDIS uses the American College of Radiology-National Electrical Manufacturers Association 2.0 protocol and the DIN-PACS uses the Digital Imaging and Communications in Medicine (DICOM) 3.0 protocol. An interface engine was required to change some data formats, as well as to address some nonstandard HL7 data being output from the CHCS. In addition, there are differences in terminology between fields and segments in all three protocols. This interface is in use at 20 military facilities throughout the world. The interface reduces the amount of manual entry into more than one automated system to the smallest level possible. Data mapping during installation saved time, improved productivity, and increased user acceptance during PACS implementation. It also resulted in more standardized database entries in both the HIS (CHCS) and the RIS (PACS). PMID- 10835135 TI - Signal intensity artifacts in clinical MR imaging. AB - Signal intensity artifacts are often encountered during magnetic resonance (MR) imaging. Occasionally, these artifacts are severe enough to degrade image quality and interfere with interpretation. Signal intensity artifacts inherent in local coil imaging include intensity gradients and local intensity shift artifact. The latter can be minimized but not eliminated with optimal coil design and tuning. Improper coil or patient positioning can produce subtle or, in some cases, severe signal intensity artifacts, and each is easily corrected. Signal intensity artifacts and image degradation can also occur in a perfectly functioning coil if protocols are not optimized. Failure of decoupling mechanisms can produce signal intensity artifacts that will not respond to protocol optimization and will worsen with gradient imaging. Improper coil tuning manifests as a shading artifact that can mimic other findings. Signal-degrading artifacts may be caused by a ferromagnetic foreign body in the imager. Signal intensity artifacts can also result from performing ultrafast imaging with coils that were not designed for this type of imaging or from MR imaging system malfunction. Familiarity with the various causes of signal intensity artifacts is necessary to maintain optimal image quality and should be required as part of any MR imaging quality assurance program. PMID- 10835136 TI - Breast implant classification with MR imaging correlation: (CME available on RSNA link) AB - Rupture is now recognized as an important and common complication of breast implants. Magnetic resonance (MR) imaging is the most accurate method for evaluating implant integrity but requires an understanding of the numerous variations in implant construction that are encountered clinically. To assist in diagnosis, the authors provide an MR-oriented breast implant classification scheme based on data from 4,014 patients (>9,966 current or previous implants), the literature, and other primary documentation. This scheme consists of 14 implant types: 1) single-lumen silicone gel-filled, 2) single-lumen gel-saline adjustable, 3) single-lumen saline-, dextran-, or polyvinyl pyrrolodone-filled, 4) standard double-lumen, 5) reverse double-lumen, 6) reverse-adjustable double lumen, 7) gel-gel double-lumen, 8) triple-lumen, 9) Cavon "cast gel", 10) custom, 11) solid pectus, 12) sponge (simple or compound), 13) sponge (adjustable), and 14) other. The MR imaging and mammographic appearance of many implant types is correlated with their actual appearance after explantation. A brief history of prosthetic breast augmentation and reconstruction is also provided to allow this classification method to be placed in historical perspective. Knowledge of the variety of breast implant types will help reduce misdiagnoses by providing imagers with better understanding of the expected appearances of breast implants. This classification scheme will allow stratification of data for studying incidence, prevalence, and risk factors for and causes of implant failure, as well as permitting better correlation with patient symptoms and surgical outcome. PMID- 10835137 TI - Thoughts for a new millennium. PMID- 10835138 TI - Legislative initiatives paid off as congress adjourned for 1999. PMID- 10835139 TI - RSV and the infant. PMID- 10835140 TI - The moral work of housekeeping issues in home care. PMID- 10835141 TI - Suicide awareness of older adults. PMID- 10835142 TI - Help may be only a phone call away for oncology patients. PMID- 10835143 TI - The future of home care. PMID- 10835144 TI - The regulation of nursing practice today. AB - The interface and complexity of licensure and scope of practice issues with professional standards of practice can be very confusing, particularly because the vary from state to state. This article provides an overview of the influences of regulatory boards, professional nursing organizations, and employers on the nursing practice. The roles of both regulatory boards and professional organizations are reviewed, including authority sources and mission differences. The employer's role is reviewed briefly, including attempts to "narrow" the licensed nurse's scope of practice. PMID- 10835145 TI - Redesigning for success: managing the prospective payment system. AB - Managed care and the new Medicare prospective pay system are dramatically changing the way home health agencies conduct their business. Agencies are busy downsizing, streamlining, and looking for creative ways to do more with less. Many industries have found success with process reengineering, the concepts of which are outlined in this article so that agencies may adapt them to "recreate" themselves. Practical measures are demonstrated through the steps one large home health agency has taken to address these issues. PMID- 10835146 TI - Home care rehabilitation standards. PMID- 10835147 TI - Psychiatric care: using all our skills. PMID- 10835148 TI - Change and growth in home care: telehealth care. PMID- 10835149 TI - Respecting a patient's autonomy: a social worker's struggle. PMID- 10835150 TI - A unique model of psychotherapy and advocacy. PMID- 10835151 TI - Medicare beneficiary access to home care decreases further. PMID- 10835152 TI - Suicide and home care: the important role of nursing. PMID- 10835153 TI - What's in a name? Lessening the confusion over drug names. AB - Confusion over the similarity of drug names, either written or spoken, accounts for approximately one-quarter of all reports to the U.S. Pharmacopeia (USP) Medication Errors Reporting (MER) program. This issue involves confusion between similar brand names, similar generic names, and similar brand and generic names. Such confusion is compounded by illegible handwriting, incomplete knowledge of drug names, newly available products, similar packaging or labeling, and incorrect selection of a similar name from a computerized product list. PMID- 10835154 TI - Psychiatric rehabilitation: a framework for psychiatric home care. AB - Psychiatric home care offers excellent opportunities to help the psychiatric patient achieve positive outcomes. Home care agencies need to develop a treatment model that supports the type of care provided to the psychiatric patient, such as psychiatric rehabilitation. This model purports that mental illness is a biologic illness affected by stress and inadequate problem-solving and coping skills. These patients have inadequate social skills, but they can improve. This article describes interactive tools and methods that the home care staff can use with such patients and their families. PMID- 10835155 TI - The geriatric depression scale. PMID- 10835156 TI - Sentinel events in home care and hospice organizations. PMID- 10835157 TI - End-of-life care: defining a society. PMID- 10835158 TI - Mental health: a report of the surgeon general. PMID- 10835159 TI - The need for an upstream ethics. PMID- 10835160 TI - RSV prophylaxis--building the case for home care. PMID- 10835161 TI - Legislators realize need to eliminate proposed 15% cut for home health. PMID- 10835162 TI - Self-management for the psychiatric patient at home. AB - Psychiatric patients in home care require support and treatment, but they also need to learn skills and self-management that will assist them after home care ends. Mental illness can be compared with any long-term illness, such as diabeteS or arthritis, in that patients should be prepared to cope with their illness through self-management. Mental illness should be considered no differently because patients also experience fluctuation in symptoms as a result of stress and the changing nature of their illnesses and will not always require intensive treatment. PMID- 10835163 TI - Hospice patient advocacy through complete documentation. AB - The hospice movement was built around patient choice and autonomy. Because of recent Medicare scrutiny, some patients are being denied the choice of hospice at the end of their lives. Hospice staff must learn how to accurately and completely document the care they provide so that patients are not denied access to care because they do not "meet the guidelines" or payment is denied as a result of inadequate or unclear documentation. PMID- 10835165 TI - Generalized convulsive epilepsy: possible mechanisms. AB - Neuronal mechanisms underlying focal convulsions and secondary generalized convulsions continue to be intensively investigated and many important pathophysiological processes are now recognized. Our understanding of primary generalized convulsions remains limited in spite of a variety of approaches. While there are well known clinical situations offering insights into some aspects of the pathophysiology, studies in animal models of generalized convulsions offer a deeper understanding of some of the processes likely to be occurring. Animal studies have indicated that generalized convulsive epilepsies are likely to be comprised of several types of convulsion and that these arise from more than one neuronal network. In particular, the cortex and brain stem can interact in various ways to lead to convulsions and may even act independently. Pharmacological agents can produce specific changes in the excitability of neurons and cause generalized convulsions, so providing models of the possible neurophysiological defects in humans. Based on electrophysiological recordings of generalized convulsions in these animal models, we also suggest that the convulsion involves subcortical mechanisms and raise the idea that primary generalized convulsive epilepsy arises out of intensified (synchronized) normal brain rhythms. Copyright 1999 Harcourt Publishers Ltd. PMID- 10835164 TI - A study of functional status among elders at two academic nursing centers. AB - Using the Older Americans Resources and Services (OARS) tool, this study examined quality of life related to functional status in 68 elderly clients at two community-based nurse-managed centers (NMCs) in California. Undergraduate and graduate nursing students administered the OARS to three cohort groups. Data from the tool were used to assess functional independence among these clients and organize and evaluate the quality of nursing care delivered at the centers. The study provides the basis for evidence-based nursing care that integrates research strategies and the nursing process. PMID- 10835166 TI - The special fields of neurological surgery. PMID- 10835167 TI - Long term results of multimodality treatment of craniopharyngioma in children. AB - The management of craniopharyngioma has been controversial for years. We review our 20 year experience in the treatment of paediatric craniopharyngioma. Twenty five patients were treated for craniopharyngioma at The Montreal Children's Hospital from 1972 to 1991. They included 15 males and 10 females whose median age was 10 years. The initial clinical manifestations were remarkable for a predominance of symptoms and signs related to intracranial hypertension, followed in frequency by visual and endocrinological deficits. Radiologically, there were five cystic craniopharyngiomas, one solid and 19 mixed. Several therapeutic approaches were used including stereotactic drainage followed by radiotherapy or radiosurgery (three cases), transsphenoidal removal (six cases) and subfrontal and/or pterional craniotomy for total (three cases) or partial (13 cases) removal, followed by radiotherapy in 10 cases. The follow up period averaged 11 years. Eight patients recurred, three after total and three after partial removal; none of these six patients had received radiotherapy. Two cases treated by stereotactic drainage recurred, one received adjuvant radiosurgery and the other conventional radiotherapy. Morbidity was lower, and quality of survival better, with more conservative approaches. More conservative approaches in the management of craniopharyngioma are reasonable alternatives for treatment. Similar rates of disease control are observed with less morbidity and better quality of survival. Copyright 1999 Harcourt Publishers Ltd. PMID- 10835168 TI - Orbital tumours in Vietnam. AB - The purpose of this study is to give an overview of orbital tumours in Vietnam from the first case operated on radically to the current management in the whole country. During this period we have kept studying and treating the disease but only 247 complete records were collected, including 139 cases operated and four cases where a biopsy was done. We would like to discuss the diagnosis and pathology and put forward some approaches to the orbit. Although orbital tumour is a pathological entity situated at the confluence of different specialties we think that the neurosurgical approach remains the most appropriate one. Copyright 1999 Harcourt Publishers Ltd. PMID- 10835169 TI - Medial cervical facetectomy for radiculopathy due to foraminal stenosis: 71 personal consecutive cases. AB - Medial cervical facetectomy performed on patients using a high speed drill and microscope represents an effective way for treatment of patients with radiculopathy due to multi-level, either uni- or bilateral, foraminal stenosis. The author presents the results of medial cervical facetectomy performed in 71 cases during a 4 year period. Medial cervical facetectomy appears to provide significant benefits to patients with radiculopathy secondary to foraminal stenosis. The procedure also appears to be superior to complete foraminotomy for relief of radiculopathy as the lateral third to half of the facet joint is still preserved and there is no instability problem. Copyright 1999 Harcourt Publishers Ltd. PMID- 10835170 TI - Intramedullary cavernous haemangiomas: clinical features, imaging diagnosis, surgical resection and outcome. AB - The diagnosis, surgical treatment and outcome of seven patients with histologically verified intramedullary cavernous haemangiomas are reported. Five males and two females, with progressive myelopathy were observed. The patients were aged between 7 and 61 years. The duration of their symptoms and signs before diagnosis varied from one month to 5 years. Magnetic resonance (MR) imaging was performed in all patients and spinal angiography was performed in five. All the patients underwent microsurgical resection of the cavernous haemangiomas. MR imaging diagnosed cavernous haemangiomas in all seven patients. T1-weighted images with or without contrast enhancement best defined the margins and size of the lesions. They also illustrated the exophytic components and the change in spinal cord surface well, and were important to surgical planning. T2-weighted images may be suboptimal due to their magnetic susceptibility artifacts. In all lesions, total removal was achieved. Postoperative neurological function improved in all patients in long term follow-up. With the advantages of MR examinations and more understanding about the pathological characteristics of intramedullary cavernous haemangiomas, total removal of the mass lesion should be attempted and the outcome is generally good. Copyright 1999 Harcourt Publishers Ltd. PMID- 10835171 TI - HIV-related primary cerebral lymphoma: the role of stereotactic biopsy. AB - Primary cerebral lymphoma (PLC) is a common cause of cerebral mass lesion in patients with the acquired immune deficiency syndrome. Its clinical features and radiological appearance are non-specific in this group of patients who are often quite debilitated at the time of presentation. We present a retrospective series of 9 cases of AIDS related PCL proven by stereotactic biopsy. There were no deaths from the biopsy and only one case in which there was worsening of the preoperative neurological condition. Although the overall outlook for these patients was poor (mean time to death 18 weeks), all but one showed some clinical response to palliative treatment with radiotherapy and steroids. In this context, we suggest that stereotactic biopsy as a means to positively establish the diagnosis prior to treatment is a safe technique to use in this group of often debilitated patients. Copyright 1999 Harcourt Publishers Ltd. PMID- 10835172 TI - Idiopathic inflammatory demyelinating diseases of the central nervous system: differentiating between acute disseminated encephalomyelitis and malignant multiple sclerosis. AB - Multiple sclerosis (MS) is the most frequent demyelinating disorder of the central nervous system (CNS). However, at presentation, it is frequently difficult to differentiate between malignant MS (MMS) and other fulminant CNS demyelinating diseases like acute disseminated encephalomyelitis (ADEM). The literature contains many case reports of ADEM but few series. We report on four representative cases of acute demyelinating diseases, together with evaluation of treatment, course and follow-up. We also present clinical, laboratory, neuropathologic, neuroimaging and data on therapeutic options, including follow up, in order to establish distinguishing characteristics of MMS and ADEM. Good clinical outcome from a postinfectious, monophasic episode, correlating with regressive demyelinating lesions on MRI, after more than 2 years differentiate best. Therapeutic efficacy, prior infection and initial MRI lesions seem to be of limited value. Despite the advances of neuroimaging and laboratory techniques, objective parameters are still missing, but findings on basic immunologic mechanisms of humoral and cellular response might provide further insight. Copyright 1999 Harcourt Publishers Ltd. PMID- 10835173 TI - Laboratory Studies. PMID- 10835175 TI - Case reports. PMID- 10835174 TI - Technical Notes. PMID- 10835179 TI - Regular Article. PMID- 10835219 TI - The probability of developing multiple sclerosis following an episode of optic neuritis. AB - One hundred and thirty cases of optic neuritis seen between 1953 and 1994 were prospectively studied. Seventy-nine (61%) developed definite multiple sclerosis. There were 19 cases of recurrent optic neuritis which, during the follow up, did not develop other signs of multiple sclerosis. Copyright 1999 Harcourt Publishers Ltd. PMID- 10835180 TI - The lamina terminalis and its role in fluid and electrolyte homeostasis. AB - The lamina terminalis, which forms most of the anterior wall of the third ventricle, consists of the median preoptic nucleus and two circumventricular organs (CVOs), the subfornical organ and organum vasculosum of the lamina terminalis. These latter two regions lack a blood-brain barrier and, unlike other regions of the brain, are influenced by the hormonal and ionic composition of the blood. The CVOs of the lamina terminalis are rich in receptors for a number of circulating peptides and the subfornical organ and the OVLT are clearly established as the prime cerebral targets for circulating angiotensin II, atrial natriuretic peptide (AVP) and relaxin to influence central nervous system pathways regulating body fluid homeostasis. Together with the median preoptic nucleus, these two CVOs also detect changes and relay neural signals relating to the tonicity of body fluids and play important roles in osmoregulatory fluid intake and excretion. The neural circuitry of the lamina terminalis involves both afferent and efferent connections to several other regions of the brain, and neurons within the individual components of lamina terminalis are reciprocally connected with each other. This neural circuitry subserves the influence that the lamina terminalis exerts on vasopressin secretion, thirst, the appetite for salt, renal sodium excretion and renin secretion by the kidney. Copyright 1999 Harcourt Publishers Ltd. PMID- 10835220 TI - New study asks: do HMOs make a difference? PMID- 10835221 TI - Biology and terrorism in an uncertain world. PMID- 10835222 TI - Bioterrorism: are we prepared? PMID- 10835223 TI - Behind the scenes at USAMRIDD. The United States Army Medical Research Institute of Infectious Diseases. PMID- 10835224 TI - Bioterrorism and the public health. AB - We were well prepared to handle the worst case scenario with the Y2K computer bug. Fortunately, our concerns were unfounded. The threat of chemical and biological terrorism, however, is real, and it is a daily threat. Physicians play a major role in the identification of events as well as their mitigation. We all hope disasters will never occur. As with most emergencies, however, sound planning and training will reduce the risks to our patients. PMID- 10835225 TI - A state prepares. PMID- 10835226 TI - The local perspective. PMID- 10835227 TI - Treating a fractured ankle over the phone. PMID- 10835229 TI - An approach for enhancing heterologous production of Providencia rettgeri penicillin acylase in Escherichia coli. AB - Heterologous production of Providencia rettgeri penicillin acylase (PAC) was optimized in Escherichia coli. Several factors, including carbon, temperature, and host effects, were identified to be critical for the enzyme overproduction. The optimum culture conditions for the enzyme production vary for different host/vector systems. With the optimization, both volumetric and specific PAC activities could be significantly improved by more than 50-fold compared to the native expression in P. rettgeri. The heterologous production could be possibly limited by translation or posttranslational steps, depending on the culture temperature and host/vector system. To our knowledge, this is the first evidence demonstrating the limiting step for the production of P. rettgeri PAC and the existence of the P. rettgeri PAC precursor. PMID- 10835228 TI - Kinetic study of the conversion of different substrates to lactic acid using Lactobacillus bulgaricus. AB - Lactic acid fermentation includes several reactions in association with the microorganism growth. A kinetic study was performed of the conversion of multiple substrates to lactic acid using Lactobacillus bulgaricus. Batch experiments were performed to study the effect of different substrates (lactose, glucose, and galactose) on the overall bioreaction rate. During the first hours of fermentation, glucose and galactose accumulated in the medium and the rate of hydrolysis of lactose to glucose and galactose was faster than the convesion of these substrates. Once the microorganism built the necessary enzymes for the substrate conversion to lactic acid, the conversion rate was higher for glucose than for galactose. The inoculum preparation was performed in such a way that healthy young cells were obtained. By using this inoculum, shorter fermentation times with very little lag phase were observed. The consumption patterns of the different substrates converted to lactic acid were studied to determine which substrate controls the overall reaction for lactic acid production. A mathematical model (unstructured Monod type) was developed to describe microorganism growth and lactic acid production. A good fit with a simple equation was obtained. It was found experimentally that the approximate ratio of cell to substrate was 1 to 10, the growth yield coefficient (Y(XS)) was 0.10 g cell/g substrate, the product yield (Y(PS)) was 0.90 g lactic acid/g substrate, and the alpha parameter in the Luedeking-Piret equation was 9. The Monod kinetic parameters were obtained. The saturation constant (K(S)) was 3.36 g/L, and the specific growth rate (microm ) was 1.14 l/h. PMID- 10835230 TI - The growth factor inhibitor suramin reduces apoptosis and cell aggregation in protein-free CHO cell batch cultures. AB - We have previously shown that Chinese hamster ovary (CHO) cells capable of growing in medium free of exogenous proteins die by apoptosis during all stages of a batch culture (Zanghi et al., 1999). On the basis of the hypothesis that extracellular death factors might be important in apoptosis under these conditions, we examined the effect of the growth factor inhibitor and antitumor agent suramin on CHO cell growth and apoptosis in serum-free culture. Suramin protected against apoptosis during exponential growth, as indicated by the absence of DNA laddering and an increase in cell viability from roughly 70% to above 95%. Suramin also effectively dispersed cell aggregates so that single-cell suspension culture was possible. However, suramin did not protect against apoptosis during the death phase, in contrast to serum, suggesting that antiapoptotic factors in the serum remain to be discovered. The increased viable cell yield following suramin supplementation resulted in a 40% increase in product yield, based on results with cells expressing recombinant secreted alkaline phosphatase. Polysulfated compounds dextran sulfate and polyvinyl sulfate worked nearly as well as suramin in dispersing cell clumps and increasing viable cell yield, which implies that suramin's high sulfate group density may be responsible for its effects in cell culture. In addition, suramin was beneficial for long-term adaptation of CHO cells to protein-free media suspension culture, and the compound was synergistic with insulin in accelerating this adaptation time. PMID- 10835231 TI - Engineering a disulfide bond in recombinant manganese peroxidase results in increased thermostability. AB - Manganese peroxidase (MnP) produced by Phanerochaete chrysosporium, which catalyzes the oxidation of Mn(2+) to Mn(3+) by hydrogen peroxide, was shown to be susceptible to thermal inactivation due to the loss of calcium [Sutherland, G. R. J.; Aust, S. D. Arch. Biochem. Biophys. 1996, 332, 128-134]. The recombinant enzyme, lacking glycosylation, was found to be more susceptible [Nie, G.; Reading, N. S.; Aust, S. D. Arch. Biochem. Biophys. 1999, 365, 328-334]. On the basis of the properties and structure of peanut peroxidase, we have engineered a disulfide bond near the distal calcium binding site of MnP by means of the double mutation A48C and A63C. The mutant enzyme had activity and spectral properties similar to those of native, glycosylated MnP. The thermostabilities of native, recombinant, and mutant MnP were studied as a function of temperature and pH. MnPA48C/A63C exhibited kinetics of inactivation similar to that of native MnP. The addition of calcium decreased the rate of thermal inactivation of the enzymes, while EGTA increased the rate of inactivation. Thermally treated MnPA48C/A63C mutant was shown to contain one calcium, and it retained a percentage of its original manganese oxidase activity; native and recombinant MnP were inactivated by the removal of calcium from the protein. PMID- 10835232 TI - Combining cell culture analogue reactor designs and PBPK models to probe mechanisms of naphthalene toxicity. AB - An alternative method of evaluating the toxicology of a chemical is to use cultured mammalian cells in a novel cell culture analogue reactor (CCA) together with a corresponding physiologically based pharmacokinetic model (PBPK). The PBPK is a mathematical model that divides the body into compartments representing organs, integrating the kinetic, thermodynamic, and anatomical parameters of the animal. The bioreactor is a physical replica of the PBPK; where the PBPK specifies an organ or tissue compartment, the bioreactor contains compartments with a corresponding cell type. The device is a continuous, dynamic system composed of multiple cell types that interact through a common circulating cell culture medium. The bioreactor and the model are coupled to evaluate the plausibility of the molecular mechanism that is input into the model. This concept is tested with naphthalene as a model of PAH (polycyclic aromatic hydrocarbons) toxicants. Two physically different CCA reactors were tested with naphthalene, and different results were observed. In the prototype system using cells attached to glass dilution bottles, naphthalene dosing resulted in generation of a circulating metabolite from the "liver" compartment (based on H4IIE cells from a rat hepatoma) that caused cell death in the "lung" compartment (L2 cells from a rat lung), as well as depletion of glutathione in the L2 cells. An improved CCA using packed bed reactors of microcarrier cultured cells did not show differences between naphthalene-dosed and nondosed controls. To explain the different responses of the two CCA designs, PBPKs of the two reactors were tested with variations in physical and kinetic parameters, and toxic mechanism. When the toxic metabolite of naphthalene was naphthoquinone rather than naphthalene epoxide as initially assumed, the PBPK results were consistent with the results of the two CCA designs. This result indicates that the mechanism of naphthalene toxicity in the CCAs may be mediated through naphthoquinone formation. The CCA PBPK concept is demonstrated to be applicable to the study of toxic mechanisms. In particular, use of this approach suggests that in vitro naphthalene toxicity is mediated through the naphthoquinone metabolite. PMID- 10835233 TI - Production and elicitation of benzalacetone and the raspberry ketone in cell suspension cultures of Rubus idaeus. AB - Production levels of p-coumaric acid (p-CA), p-hydroxyphenylbut-3-ene-2-one (benzalacetone), and p-hydroxyphenyl-2-butanone (raspberry ketone) were measured in raspberry cell suspension cultures to investigate metabolite dynamics in a short (two-step) pathway. Intracellular concentrations of benzalacetone and the raspberry ketone fluctuated during the time course of a normal batch culture cycle but showed higher levels during periods of rapid growth. Cells elicited with the signal coupler methyl jasmonate yielded a 2- to 3-fold increase in metabolite concentrations after 24 h. The results suggest that raspberry ketone production is rapidly inducible during periods of high carbohydrate utilization. It is not an end product, however, and undergoes conversion to subsequent metabolites. PMID- 10835234 TI - Modeling retrovirus production for gene therapy. 2. Integrated optimization of bioreaction and downstream processing. AB - In this work a model envisaging the integrated optimization of bioreaction and downstream processing is presented. This model extends the work presented in part 1 of this pair of papers by adding ultrafiltration to process optimization. The new operational parameters include ultrafiltration time, pressure, and stirring rate. For global optimization, the model uses as constraints the final product titer and quality to be achieved after downstream processing. This extended model was validated with the same system used in part 1, i.e., PA317 cells producing a recombinant retrovirus containing the LacZ gene as a marker in stirred tanks using porous supports. Optimization of the extended model led to the conclusion that bioreaction should have two steps, batch and perfusion, similar to what was found in part 1. Ultrafiltration in a stirred cell should be performed at low pressures and stirring rates to reduce the losses of infective retroviruses. Sensitivity analysis performed on the results of the integrated optimization showed that under optimal conditions the productivity is less sensitive to the parameters related to ultrafiltration than to those associated with bioreaction. These results were interpreted as reflecting the high yield of ultrafiltration (90%). The relevance of the model extension to perform integrated optimization was also demonstrated since a restriction in the specific ultrafiltration area in downstream processing conditioned perfusion duration and perfusion rate in bioreaction. This clearly indicates that overall process optimization cannot be achieved without integrated optimization. PMID- 10835235 TI - Study of vitamin ester synthesis by lipase-catalyzed transesterification in organic media. AB - Immobilized lipase from Candida antarctica (Novozym 435) was used in organic media to catalyze esterifications of vitamins (ascorbic acid and retinol) from hydroxy acid. We described the synthesis of retinyl L-lactate by transesterification between retinol and L-methyl lactate with yield reaching 90% and the synthesis of ascorbyl L-lactate by transesterification between ascorbic acid and L-methyl lactate with yield reaching 80%. The kinetic study of the esterification of vitamins with L-methyl lactate in organic media has been carried out and agrees with ping-pong-ordered Bi-Bi when the initial vitamin concentration is low. When initial vitamin concentration is high, the kinetic is similar to a hybrid ping-pong-ordered Bi Bi or hybrid ping-pong-random Bi Bi mechanism. However, with high initial substrate concentration, change of the kinetic by other phenomena, such as interaction of substrates with molecular sieves, adsorption of the methanol formed, and decreases of substrate diffusion, could be considered. It is obvious that in these conditions, classical enzymology (i.e., Michaelian enzymology) cannot be used for the interpretation of results. PMID- 10835236 TI - Biocatalytic transformation of [(2-Hydroxyethyl)thio]acetic acid and thiodiglycolic acid from thiodiglycol by Alcaligenes xylosoxydans ssp. xylosoxydans (SH91). AB - A Gram-negative bacterium, Alcaligenes xylosoxydans ssp. xylosoxydans (SH91), consumed thiodiglycol (TDG), the nontoxic hydrolysis product of sulfur mustard, as a primary carbon source and transformed TDG to commercially relevant chemical precursors, [(2-hydroxyethyl)thio]acetic acid (HETA) and thiodiglycolic acid (TDGA). Aerobic fed batch and repeated batch experiments were run to compare the molar yields of HETA and TDGA that result under different operating policies. In repeated batch experiments, 35% of the TDG was converted to HETA. Under the conventional batch process and a repeated fed batch process, the HETA yields were reduced (21% and 18%, respectively), while the yield of TDGA was increased (47% and 31%,respectively). This work demonstrated that cell growth associated biocatalytic transformations were manipulated to achieve a desired byproducts profile through an understanding of the specific reaction and cell growth kinetics and by altering the reaction operating policy accordingly. PMID- 10835237 TI - Activation and stabilization of penicillin V acylase from streptomyces lavendulae in the presence of glycerol and glycols. AB - Penicillin V acylase (EC 3.5.1.11) from Streptomyces lavendulae showed both enhanced activity and stability in mixed water/glycerol and water/glycols solvents. The catalytic activity was increased up to a critical concentration of these cosolvents, but further addition of the latter led to a gradual protein deactivation. The highest stabilizing effect was achieved in the presence of glycerol. Thermal stability was increased proportionally to the concentration of glycerol and glycols in the reaction mixture only if the amount added is below the threshold concentration. Reaction conditions that allow simultaneously enhanced activity and stability in the hydrolysis of penicillin V catalyzed by penicillin V acylase from S. lavendulae could be established. PMID- 10835238 TI - Comparative fatty acid selectivity of lipases in esterification reactions with glycerol and diol analogues in organic media. AB - Reaction selectivity of Pseudomonas cepacia, Rhizomucor miehei, and Candida antarctica B lipases was assessed in multicompetitive esterification reaction mixtures containing an homologous series of n-chain even carbon number fatty acid (FA; C4-C18) substrates and a single alcohol cosubstrate (glycerol, 1,2 propanediol (1,2-PD), or 1, 3-propanediol (1,3-PD)) in tert-butyl methyl ether at water activity of 0.69 or 0.90 and a reaction temperature of 35 degrees C. For P. cepacia lipase, the ordinal patterns of FA selectivities observed were, with glycerol, C8 > C10, C6, C16 > other FA; with 1,2-PD and 1, 3-PD, C16 > C8 > C14 > other FA. For R. miehei lipase, the ordinal patterns of FA selectivities observed were, with glycerol, C8 > C12 > C10, C14 > other FA; with 1,2-PD and 1,3-PD, C8 > C12 > other FA. For C. antarctica B lipase, the ordinal patterns of FA selectivities observed were, with glycerol, C8 > C10, C6, C12 > other FA; with 1, 2-PD, C8 > C10, C6 > other FA; and with 1,3-PD, C8 > C10 > C6 > other FA. The differences in selectivity among FA ranged up to 16-fold, depending upon the lipase and alcohol cosubstrate used. These findings represent intrinsic and substrate-modulated features of FA selectivities that are of particular relevance to the use of lipases for acylglycerol synthesis reactions. PMID- 10835239 TI - Toluene removal in an automated cyclical bioreactor. AB - A control scheme was developed for the automation of toluene removal in a cyclical bioreactor. Toluene was added to the self-cycling fermentor by diffusion across a silicone membrane. Transient dissolved oxygen, carbon dioxide evolution, and oxidation-reduction potential (ORP) were screened as potential control variables. Through experimentation, ORP was deemed most effective. Control algorithms based on real-time estimates of the first and second derivatives of the ORP signal were tested. Although both approaches resulted in stable operation of the reactor, average toluene removal efficiencies of 95% were realized when control was based on the second derivative. This was significantly higher than the 77% efficiencies obtained when the control scheme centered on the first derivative of the transient ORP signal. The system developed was self-regulating, ensuring that a high toluene removal rate, on the order of 1.1 g h(-1), was maintained from cycle to cycle. PMID- 10835240 TI - Prolonging cell-free protein synthesis by selective reagent additions. AB - Factors causing the early cessation of protein synthesis have been studied in a cell-free system from Escherichia coli. We discovered that phosphoenol pyruvate (PEP), the secondary energy source for ATP regeneration, and several amino acids are rapidly degraded during the cell-free protein synthesis reaction. The degradation of such compounds takes place even in the absence of protein synthesis. This degradation severely reduces the capacity for protein synthesis. The lost potency was completely recovered when the reaction mixture was supplied with additional PEP and amino acids. Of the 20 amino acids, only arginine, cysteine, and tryptophan were required to restore system activity. Through repeated additions of PEP, arginine, cysteine,and tryptophan, the duration of protein synthesis was greatly extended. In this fed-batch reaction, after a 2 h incubation, the level of cell-free synthesized chloramphenicol acetyl transferase (CAT) reached 350 microg/mL, which is 3.5 times the yield of the batch reaction. Addition of fresh magnesium further extended the protein synthesis. As a result, through coordinated additions of PEP, arginine, cysteine, tryptophan, and magnesium, the final concentration of cell-free synthesized CAT increased more than 4-fold compared to a batch reaction. SDS-PAGE analysis of such a fed-batch reaction produced an obvious band of CAT upon Coomassie Blue staining. PMID- 10835241 TI - Hairy root culture in a liquid-dispersed bioreactor: characterization of spatial heterogeneity. AB - A liquid-dispersed reactor equipped with a vertical mesh cylinder for inoculum support was developed for culture of Atropa belladonna hairy roots. The working volume of the culture vessel was 4.4 L with an aspect ratio of 1.7. Medium was dispersed as a spray onto the top of the root bed, and the roots grew radially outward from the central mesh cylinder to the vessel wall. Significant benefits in terms of liquid drainage and reduced interstitial liquid holdup were obtained using a vertical rather than horizontal support structure for the biomass and by operating the reactor with cocurrent air and liquid flow. With root growth, a pattern of spatial heterogeneity developed in the vessel. Higher local biomass densities, lower volumes of interstitial liquid, lower sugar concentrations, and higher root atropine contents were found in the upper sections of the root bed compared with the lower sections, suggesting a greater level of metabolic activity toward the top of the reactor. Although gas-liquid oxygen transfer to the spray droplets was very rapid, there was evidence of significant oxygen limitations in the reactor. Substantial volumes of non-free-draining interstitial liquid accumulated in the root bed. Roots near the bottom of the vessel trapped up to 3-4 times their own weight in liquid, thus eliminating the advantages of improved contact with the gas phase offered by liquid-dispersed culture systems. Local nutrient and product concentrations in the non-free-draining liquid were significantly different from those in the bulk medium, indicating poor liquid mixing within the root bed. Oxygen enrichment of the gas phase improved neither growth nor atropine production, highlighting the greater importance of liquid solid compared with gas-liquid oxygen transfer resistance. The absence of mechanical or pneumatic agitation and the tendency of the root bed to accumulate liquid and impede drainage were identified as the major limitations to reactor performance. Improved reactor operating strategies and selection or development of root lines offering minimal resistance to liquid flow and low liquid retention characteristics are possible solutions to these problems. PMID- 10835242 TI - Rapid expansion from supercritical to aqueous solution to produce submicron suspensions of water-insoluble drugs. AB - Stable suspensions of submicron particles of cyclosporine, a water-insoluble drug, have been produced by rapid expansion from supercritical to aqueous solution (RESAS). To minimize growth of the cyclosporine particles, which would otherwise occur in the free jet expansion, the solution was sprayed into an aqueous Tween-80 (Polysorbate-80) solution. Steric stabilization by the surfactant impedes particle growth and agglomeration. The particles were an order of magnitude smaller than those produced by RESS into air without the surfactant solution. Concentrations as high as 38 mg/mL for 400-700 nm particles were achieved in a 5.0% (w/w) Tween-80 solution. PMID- 10835243 TI - New polymers forming aqueous two-phase polymer systems. AB - A new type of polymer, starch-modified by acrylamide, has been developed for application in aqueous two-phase systems (ATPS) for protein separation. Partial hydrolysis and acrylamide modification of starch to different degrees make it suitable for forming ATPS with poly(ethylene glycol) in a moderate concentration range. The potential of the polymer to form ATPS with the thermoprecipitating copolymer of 1-vinylimidazole with N-vinylcaprolactam (poly-VI/VCL) has been evaluated. The thermoprecipitation properties of poly-VI/VCL and Cu(II)-loaded poly-VI/VCL have been studied for application in metal affinity partitioning. The formation of ATPS with Cu(II)-loaded thermoprecipitating copolymer was critically achieved for poly-VI/VCL (10/90) copolymer in under-loaded metal concentrations. With the Cu(II)-loaded copolymer, poly-VI/VCL in the top phase and modified starch in the bottom phase, the ATPS formed was used for the purification of alpha-amylase inhibitor from wheat meal. The protein partitioned in the top phase and phase-separated polymer-protein complex could be precipitated by salt. The protein inhibitor was recovered with a yield of 75%. PMID- 10835244 TI - Studies on the batch adsorption of plasmid DNA onto anion-exchange chromatographic supports. AB - The adsorption of a supercoiled 4.8 kbp plasmid onto quaternary ammonium anion exchangers was studied in a finite bath. Equilibrium experiments were performed with pure plasmid, at 25 degrees C, using commercial Q-Sepharose matrices differing in particle diameter (High Performance, 34 microm; Fast Flow, 90 microm; and Big Beads, 200 microm) and a recently commercialized ion-exchanger, Streamline QXL (d(p) = 200 microm) at different salt concentrations (0.5, 0.7, and 1 M NaCl). Plasmid adsorption was found to follow second-order kinetics (Langmuir isotherm) with average association constants K(A) = 0.32+/-0.12 mL microg(-)(1) and K(A) = 0.25+/-0.15 mL microg(-1) at 0.5 and 0.7 M Nacl, respectively. The maximum binding capacities were not dependent on the ionic strength in the range 0.5-0.7 M but decreased with increasing particle diameter, suggesting that adsorption mainly occurs at the surface of the particles. No adsorption was found at 1 M NaCl. A nonporous model was applied to describe the uptake rate of plasmid onto Streamline QXL at 0.5 M NaCl. The overall process rate was controlled by mass transfer in the regions of low relative amounts of adsorbent (initial stages) and kinetically controlled in the later stages of the process for high relative amounts of adsorbent. The forward reaction rate constant (k(1) = 0.09+/-0.01 mL mg(-1) s(-1)) and film mass transfer coefficient (K(f) = (6 +/- 2) x 10(-4) cm s(-1)) were calculated. Simulations were performed to study the effect of the relative amount of adsorbent on the overall process rate, yield, and media capacity utilization. PMID- 10835245 TI - Development and qualification of a novel virus removal filter for cell culture applications. AB - Commercial bioreactors employing mammalian cell cultures to express biological or pharmaceutical products can become contaminated with adventitious viruses. The high expense of such a contamination can be reduced by passing all gases and fluids feeding the bioreactor through virus inactivation or removal steps, which act as viral barriers around the bioreactor. A novel virus barrier filter has been developed for removing viruses from serum-free cell culture media. This filter removes the 20 nm minute virus of mice by >3 log reduction value (LRV), the 28 nm bacteriophage PhiX174 by >4.5 LRV, the mycoplasma Acholeplasma laidlawii by > or =8.8 LRV, and the bacteria Brevundimonas diminuta by > or =9.2 LRV. Robust removal occurs primarily by size exclusion as demonstrated over a wide range of feedstocks and operating conditions. The filtered media are indistinguishable from unfiltered media in growth of cells to high densities, maintenance of cell viability, and productivity in expressing protein product. Insulin and transferrin show high passage through the filter. The virus barrier filter can be autoclaved. The relatively high membrane permeability enables the use of a moderate filtration area. PMID- 10835246 TI - Process options in hepatitis B surface antigen extraction from transgenic potato. AB - The process conditions for recombinant hepatitis B surface antigen (HBsAg) extraction from transgenic potato were examined. The effects of temperature, the reducing agent beta-mercaptoethanol (BME), and proteinase inhibitors on the level of antigenic activity of recovered HBsAg were determined. Sedimentation profiles were performed to characterize HBsAg assembly into virus-like particles. Increasing the temperature of the sample for about 1 min increased the measured HBsAg antigenic activity. The optimum temperature was around 50 degrees C. A 3 fold enhancement of the antigenic activity was obtained in extract from transgenic potato expressing HBsAg, when monoclonal antibodies were used to assay for HBsAg. When antigenic activity was determined by polyclonal antibodies, no enhancement in the antigenic activity was obtained. Temperature may affect the conformation of the a epitope to which the monoclonal antibodies bind or alter the fluidity of surface lipid regions. BME increased the antigenic activity of HBsAg up to 4-fold when monoclonal antibodies directed against the a determinant were used, but there was no increase with polyclonal antibodies. This observation suggests that BME affects the structure or presentation of the a epitope. In the presence of BME and leupeptin, a proteinase inhibitor, higher antigenic activity was obtained. Leupeptin might protect the antigen, which might become more susceptible to proteolytic degradation after reduction, as a result of stimulation of sulfhydryl proteases. Although both temperature and BME increased the antigenic activity of HBsAg individually, when combined their interaction was antagonistic, resulting in reduced antigenic activity. Different proteinase inhibitors, including leupeptin, aprotinin, E-64, pefabloc, and pepstatin, had no significant effect on HBsAg from potato extract in a 2 h period in the absence of BME. The sedimentation profile of potato-produced HBsAg was determined in 5-30% sucrose gradients. Yeast-derived recombinant HBsAg was used as a positive control. The HBsAg from transgenic potato showed sedimentation and density properties that are very similar to the yeast-produced antigen, indicating assembly into virus-like particles. BME treatment did not change the sedimentation profile. PMID- 10835247 TI - Affinity purification of fusion chaperonin Cpn60-(His)(6) from thermophilic bacterium Bacillus strain MS and its use in facilitating protein refolding and preventing heat denaturation. AB - The cpn60 gene from Bacillus strain MS, which is highly homologous to Bacillus stearothermophilus, was cloned. Cpn60 with a hexahistidine affinity tag (His)(6) fused to its C-terminus (cpn60-(His)(6)) was overproduced in Escherichia coli. Cpn60-(His)(6) was expressed in a soluble form in E. coli. and purified to homogeneity in a single step by nickel chelate affinity chromatography. Cpn60 (His)(6) formed a tetradecamer and had ATPase activity. Cpn60-(His)(6) mediated refolding of guanidine hydrochloride unfolded pig heart malic dehydrogenase (MDH) and Thermus flavus MDH at 25 and 70 degrees C, respectively, in an ATP-dependent manner. In addition, cpn60-(His)(6) prevented heat denaturation of pig heart MDH and T. flavus MDH at 30 and 80 degrees C, respectively, in an ATP-dependent manner. Therefore, cpn60-(His)(6) facilitates protein refolding and prevents heat denaturation of proteins across a wide temperature range. PMID- 10835248 TI - Modeling conductive heat transfer during high-pressure thawing processes: determination of latent heat as a function of pressure. AB - A numerical heat transfer model for predicting product temperature profiles during high-pressure thawing processes was recently proposed by the authors. In the present work, the predictive capacity of the model was considerably improved by taking into account the pressure dependence of the latent heat of the product that was used (Tylose). The effect of pressure on the latent heat of Tylose was experimentally determined by a series of freezing experiments conducted at different pressure levels. By combining a numerical heat transfer model for freezing processes with a least sum of squares optimization procedure, the corresponding latent heat at each pressure level was estimated, and the obtained pressure relation was incorporated in the original high-pressure thawing model. Excellent agreement with the experimental temperature profiles for both high pressure freezing and thawing was observed. PMID- 10835249 TI - Multilayer binding of proteins to polymer chains grafted onto porous hollow-fiber membranes containing different anion-exchange groups. AB - Various anion-exchange groups were introduced into the polymer chains grafted onto a porous hollow-fiber membrane for protein recovery by radiation-induced graft polymerization and subsequent functionalization of a monomer containing an epoxy group. The graft chains extended from the pore surface toward the pore interior, resulting in the multilayer binding of proteins to the graft chains. Combinations of three anion-exchange groups, namely, amino (AM), ethylamino (EA), and diethylamino (DEA) groups, and three proteins, namely, beta-lactoglobulin, bovine serum albumin, and urease, were examined to evaluate the degree of multilayer binding of protein to the graft chains in the permeation mode. Multilayer binding was observed for hollow-fiber membranes containing EA and DEA groups, with conversions of epoxy groups to EA or DEA groups of higher than 80%. The amount of adsorbed protein remained constant irrespective of the conversion for the hollow-fiber membrane containing an AM group. The dependence of the flux on the conversion was consistent with that of the degree of multilayer binding to the graft chains. PMID- 10835250 TI - Comparisons of the glycosylation of a monoclonal antibody produced under nominally identical cell culture conditions in two different bioreactors. AB - The murine B-lymphocyte hybridoma cell line, CC9C10, was grown in serum-free continuous culture at steady-state dissolved oxygen (DO) concentrations of 10%, 50%, and 100% of air saturation in both LH Series 210 (LH) and New Brunswick Scientific (NBS) CelliGen bioreactors. All culture parameters were monitored and controlled and were nominally identical at steady state in the two bioreactors. The secreted monoclonal antibody (mAb), an immunoglobulin G(1), was purified and subjected to enzymatic deglycosylation using peptide N-glycosidase F (PNGase F). Asparagine-linked (N-linked) oligosaccharide pools released from mAb samples cultured in each bioreactor at each of the three DO setpoints were analyzed by high-pH anion-exchange chromatography with pulsed amperometric detection (HPAEC PAD). The predominant N-linked structures were core-fucosylated asialo biantennary chains with varying galactosylation. There were also minor amounts of monosialyl oligosaccharides and trace amounts of afucosyl oligosaccharides. The level of DO affects the glycosylation of this mAb. A definite reduction in the level of galactosylation of N-glycan chains was observed at lower DO in both bioreactors, as evidenced by prominent increases in the relative amounts of agalactosyl chains and decreases in the relative amounts of digalactosyl chains with the relative amounts of monogalactosyl chains being comparatively constant. However, the quantitative results are not precise matches between the two bioreactors. The effect of DO on galactosylation is less pronounced in the NBS bioreactor than in the LH bioreactor, particularly the shift between the relative amounts of agalactosyl and digalactosyl chains in 10% and 50% DO. There are also perceptibly higher levels of sialylation of the mAb glycans in the NBS bioreactor than in the LH bioreactor at all three DO setpoints. The results indicate that the DO effect is not bioreactor specific and that nominally identical steady state conditions in different chemostat bioreactors may still lead to some incongruities in glycosylation, possibly due to the particular architectures of the bioreactors and the design of their respective monitoring and control systems. The observed differences in N-linked glycosylation of the mAb secreted by the hybridoma grown in the LH and NBS bioreactors may be explained by the differences in oxygen supply and control strategies between the two bioreactors. PMID- 10835251 TI - Characterization of a perfusion reactor utilizing mammalian cells on microcarrier beads. AB - Our overall objective is to develop a cell culture analogue bioreactor (CCA) that can be used together with a corresponding physiologically based pharmacokinetic model (PBPK) to evaluate molecular mechanisms of toxicity. The PBPK is a mathematical model that divides the body into compartments representing organs, integrating the kinetic, thermodynamic, and anatomical parameters of the animal. The CCA bioreactor is a physical replica of the PBPK; where the PBPK specifies organs, the CCA bioreactor contains compartments with a corresponding cell type that mimics some of the characteristic metabolism of that organ. The device is a continuous, dynamic system composed of multiple cell types that interact through a common circulating cell culture medium. The CCA bioreactor and the model can be coupled to evaluate the plausibility of the molecular mechanism that is input into the model. This paper focuses on the design, development, and characterization of a CCA bioreactor to be used in naphthalene dose response studies. A CCA bioreactor prototype developed previously is improved upon by culturing the cells on microcarrier beads. Microcarrier beads with cells attached can form packed beds with cell culture medium perfusing the beds. In this study, two packed beds of cells, one with L2 cells (rat lung) and one with H4IIE cells (rat hepatoma), are linked in a physiologically relevant arrangement by a common recirculating cell culture medium. Studies of this CCA bioreactor presented here include mixing profiles, effect of reactor environment on cell viability and intracellular glutathione, naphthalene distribution profile, and initial naphthalene dosing studies. Unlike the prototype system there is no detectable response to naphthalene addition; in a companion paper we show that this discrepancy can be explained by differences in liquid residence times in the organ compartments. The perfusion reactor design is shown to have significant operating improvements over prototype designs. PMID- 10835252 TI - Hydrocolloid coating of Xenopus laevis embryos. AB - A novel technology for coating single cells and embryos with thin hydrocolloid (water-soluble polymer) films has been invented and patented. Coating is different from entrapment and immobilization in that the coating around the cell is thinner, comprising only a small fraction of the cell or embryo's diameter. Xenopus laevis embryos were coated with thin films of low-methoxy pectin (LMP), alginate, and iota- and kappa-carrageenans. These gums have different compositions and structures and as such created different coatings around the fertilized cells. All coated embryos appeared to develop normally, similar to noncoated embryos. Elemental detection by ICP-AES spectroscopy revealed that the embryo can control the diffusion of excess ions to which it is exposed during the coating process. The coatings delayed hatching by 18-24 h. Consequently, at hatch the embryos were at a more developed stage than their noncoated counterparts. The hydrocolloid coating reduced the thickness of the natural jelly coating (JC). With the iota-carrageenan coating, percent hatch was maximal, while with LMP it was minimal, as a result of the films' mechanical properties and thicknesses. LMP and alginate created smoother coatings than the carrageenans. Potential interactions between the coating and the natural JC are hypothesized. Overall, coatings appear to be a suitable tool for laboratories interested in performing longer-term experiments with embryos. PMID- 10835253 TI - Low-frequency sonophoresis: a noninvasive method of drug delivery and diagnostics. AB - Transdermal drug delivery offers an attractive alternative to injections and oral medications. However, applications of transdermal drug delivery are limited to only a few drugs as a result of low skin permeability. Application of low frequency ultrasound enhances skin permeability, a phenomenon referred to as low frequency sonophoresis. In this method, a short application of ultrasound is used to permeabilize skin for a prolonged period of time. During this period, ultrasonically permeabilized skin may be utilized for drug delivery. In addition, a sample of interstitial fluid or its components may be extracted through permeabilized skin for diagnostic applications. In this paper, we report our in vivo studies that demonstrate the principles of both of these concepts. Detailed studies on drug delivery are performed using inulin and mannitol as model drugs. Studies on diagnostics are performed using glucose as a model analyte. Applications of this technology to drug delivery and diagnostics are discussed. PMID- 10835254 TI - Effect of protease inhibitors on yield of HSV-1-based viral vectors. AB - The ability to obtain high titer replication-defective herpes simplex virus (HSV) recombinant vectors will dramatically affect their use in gene therapy clinical trials. A variety of techniques and reagents have been employed to increase the overall yield of the vector. The effects of protease inhibitors on the yield of an HSV-1-based viral vector were examined. Experiments were conducted using a commercial protease inhibitor cocktail typically used in mammalian cell culture for protein production. Contrary to our expectation for enhanced vector yield, the results showed a dramatic reduction in vector yield. Moreover, it was found that AEBSF is the only component in the protease cocktail responsible for the low vector yield. On the basis of our hypothesis regarding the mode of action of AEBSF, we suggest that it should not be included in protease inhibitor cocktails designed for use in cultures aimed at production of viral vectors derived from HSV-1 or possibly several other vectors. PMID- 10835255 TI - Alginate coating of Xenopus laevis embryos. AB - Xenopus laevis eggs were coated, immediately after squeeze-stripping and fertilization, with a thin layer ( approximately 50 microm) of film based on one of three different types of alginates which varied in their mannuronic/guluronic acid ratio. The alginate was cross-linked with either Ca or Ba ions at three different concentrations. The developmental, survival, and hatching of these embryos and the swelling of their natural jelly coats or hydrocolloid coatings were studied over 7 days, while embryos were maintained in flowing aerated water at a ratio of 85 mL per embryo or at a very diminished ratio of 1.6 mL of sterile or nonsterile MMR solution per embryo. All experiments were conducted in triplicate at 20+/-1 degrees C. Oxygen was monitored continuously. Mineral content was determined in the alginate-jelly coat and within the embryos over time. The coating conferred major advantages when the ratio between the embryos and the surrounding medium was at a minimum under nonsterile conditions, perhaps as a result of the film's resistance to diffusion. In the studied systems, the coating seemed to postpone embryo hatching to a more developed stage. In addition, the coating served as a barrier to microbial contamination and thus improved survival prospects. PMID- 10835256 TI - Optimization of ion-exchange protein separations using a vector quantizing neural network. AB - In this work, a previously proposed methodology for the optimization of analytical scale protein separations using ion-exchange chromatography is subjected to two challenging case studies. The optimization methodology uses a Doehlert shell design for design of experiments and a novel criteria function to rank chromatograms in order of desirability. This chromatographic optimization function (COF) accounts for the separation between neighboring peaks, the total number of peaks eluted, and total analysis time. The COF is penalized when undesirable peak geometries (i.e., skewed and/or shouldered peaks) are present as determined by a vector quantizing neural network. Results of the COF analysis are fit to a quadratic response model, which is optimized with respect to the optimization variables using an advanced Nelder and Mead simplex algorithm. The optimization methodology is tested on two case study sample mixtures, the first of which is composed of equal parts of lysozyme, conalbumin, bovine serum albumin, and transferrin, and the second of which contains equal parts of conalbumin, bovine serum albumin, tranferrin, beta-lactoglobulin, insulin, and alpha -chymotrypsinogen A. Mobile-phase pH and gradient length are optimized to achieve baseline resolution of all solutes for both case studies in acceptably short analysis times, thus demonstrating the usefulness of the empirical optimization methodology. PMID- 10835257 TI - Enzymatically and combinatorially generated array-based polyphenol metal ion sensor. AB - Phenolic polymers were synthesized via soybean hull peroxidase catalysis and used as metal-based sensor components in a polymer array. A sensor array for Fe(3+), Cu(2+), Co(2+), and Ni(2+) has been developed consisting of 15 phenolic homopolymers and copolymers generated from five phenolic monomers by peroxidase catalyzed oxidative polymerization. Sensing was based on the change of intrinsic polyphenol fluorescence upon addition of a metal ion or a metal ion mixture to an aqueous suspension of a polyphenol. Importantly, the fluorescence response of copolymers differed, in some cases dramatically, from the constituent homopolymers and was dependent upon the relative ratio of monomers that comprise the polymer. This finding suggests that an extremely broad range of sensor arrays can be generated from a limited number of phenolic monomers. Using a statistical analysis, histograms constructed for the four different metal ions yielded unique fingerprints of the array response and can be used to identify specific metal ions. PMID- 10835258 TI - A trimmed viral cap-independent translation enhancing sequence for rapid in vitro gene expression. AB - We prepared a short (29 nucleotides) 5' UTR that enhanced cap-independent translation in a wheat germ translation system by trimming the tobacco etch virus 5' UTR. The trimmed sequence, designated as TE(37-65), was obtained from a conserved region among several potyviruses. The productivities of uncapped reporter mRNAs carrying the TE(37-65) sequence were comparable to those of capped counterparts, in that 5-20 microg of proteins were synthesized per 1 mL of translation reaction mixture. The ribosome that entered onto the TE(37-65) sequence precisely initiated polypeptide synthesis at the defined initiation codon, which ensures rapid and efficient protein truncation analyses. Moreover, the TE(37-65) sequence is short enough to be involved in a PCR primer, which allows a simple method for rapid gene expression, i.e., PCR amplification of a target gene and succeeding in vitro transcription and translation. As a demonstration, the rapid in vitro expression of rice cDNAs using the TE(37-65) sequence was also performed. PMID- 10835259 TI - The production of hemicellulases by aerobic fungi on medium containing residues of banana plant as substrate. AB - Trichoderma harzianum strains T4 and T6, Acrophialophora nainiana, and Humicola grisea var. thermoidea were screened for their ability to produce carbohydrate degrading enzyme activities in a medium containing banana plant residue as the carbon source. The best balance of enzyme activities was obtained from cultures of H. grisea var. thermoidea. Xylanase activity from crude extract of A. nainiana had a maximum activity at pH 5.5-7.0 and a temperature range of 50-55 degrees C. It was stable up to 55 degrees C at pH 7.0 for at least 2 h. The fungi were also able to produce xylanase and pectinase activities when grown on extractives as substrate. PMID- 10835260 TI - Human remains from Blombos Cave, South Africa: (1997-1998 excavations). AB - The Middle Stone Age (MSA) layers at Blombos Cave contain abundant bifacial Still Bay points, formal and ad hoc bone artefacts, and an intentionally incised bone piece. These artefacts add weight to arguments that some aspects of modern human behavior developed earlier in sub-Saharan Africa than elsewhere. Four human teeth were recovered from the MSA strata at Blombos during the 1997-1998 excavations. Two are heavily worn deciduous teeth, and two are incomplete permanent premolar crowns. The Blombos di(1)is comparatively large in relation to modern African homologues, falling within the lower part of the observed Neandertal range. The dm(1)and P(3)are comparable to modern teeth and smaller than most Neandertal crowns. The premolars preserve horizontal circum-cervical striae that suggest palliative toothpick use. The di(1)evinces labial scratches that resemble neither the "cutmarks" that have been observed on Neandertal incisors, nor the striae that have been recorded on modern human teeth. PMID- 10835261 TI - Estimating stature in fossil hominids: which regression model and reference sample to use? AB - coResearchers have long appreciated the significant relationship between body size and an animal's overall adaptive strategy and life history. However, much more emphasis has been placed on interpreting body size than on the actual calculation of it. One measure of size that is especially important for human evolutionary studies is stature. Despite a long history of investigation, stature estimation remains plagued by two methodological problems: (1) the choice of the statistical estimator, and (2) the choice of the reference population from which to derive the parameters. This work addresses both of these problems in estimating stature for fossil hominids, with special reference to A.L. 288-1 (Australopithecus afarensis) and WT 15000 (Homo erectus). Three reference samples of known stature with maximum humerus and femur lengths are used in this study: a large (n=2209) human sample from North America, a smaller sample of modern human pygmies (n=19) from Africa, and a sample of wild-collected African great apes (n=85). Five regression techniques are used to estimate stature in the fossil hominids using both univariate and multivariate parameters derived from the reference samples: classical calibration, inverse calibration, major axis, reduced major axis and the zero-intercept ratio model. We also explore a new diagnostic to test extrapolation and allometric differences with multivariate data, and we calculate 95% confidence intervals to examine the range of variation in estimates for A.L. 288-1, WT 15000 and the new Bouri hominid (contemporary with [corrected] Australopithecus garhi). Results frequently vary depending on whether the data are univariate or multivariate. Unique limb proportions and fragmented remains complicate the choice of estimator. We are usually left in the end with the classical calibrator as the best choice. It is the maximum likelihood estimator that performs best overall, especially in scenarios where extrapolation occurs away from the mean of the reference sample. The new diagnostic appears to be a quick and efficient way to determine at the outset whether extrapolation exists in size and/or shape of the long bones between the reference sample and the target specimen. PMID- 10835262 TI - The environmental contexts of early human occupation of Georgia (Transcaucasia). AB - The hominid mandible and a third metatarsal found in Dmanisi (Republic of Georgia) are accompanied by a rich faunal assemblage and a core-chopper stone tool industry. The mandible represents a somewhat isolated morphological type of Homo erectus that appears, given the combination of its primitive and advanced traits and specific dental morphology, to be a forerunner of both late H. erectus and early archaic H. sapiens. The faunal assemblage mostly consists of Villafranchian mammals, with the majority of the species assigned to an early phase of the Upper Villafranchian (Late Villanian and Early Biharian). Faunal and paleobotanical evidence as well as the depositional nature of the site indicate that hominid occupation took place in a mosaic environment of open steppe and gallery forests. Both the concentration of resources and the warm climatic conditions in the Dmanisi region at the beginning of the early Pleistocene were favorable for hominid occupation. It is possible that hominids reached the Caucasus through the Levantine corridor, and that the environment of this region allowed them to establish a stronghold and later colonize adjacent areas. PMID- 10835263 TI - The current state of korean paleoanthropology. AB - The hominid fossil and Paleolithic archaeology records from the Korean Peninsula are extensive, but relatively little is known about the Korean human evolutionary record outside this region. The Korean paleoanthropological record is reviewed here in light of major research issues, including the hominid fossil record, relative and chronometric dating, lithic analysis, hominid subsistence, and the presence of bone tools, art and symbolism. Some of the major conclusions drawn from this review include: (1) hominid fossils have been found in nine separate sites on the Korean Peninsula; (2) possible Homo erectus fossils are present in North Korea; (3) Ryonggok Cave, in North Korea, has exposed the remains of at least five archaic Homo sapiens individuals; (4) a possible burial of an anatomically modern Homo sapiens child, discovered in Hungsu Cave in South Korea, has been tentatively dated to roughly 40,000 years ago; (5) handaxes and cleavers have been found at a number of sites near Chongokni and they appear to date to at least 100,000 years ago; and (6) taphonomic studies are necessary for addressing issues related to determining the nature of hominid-carnivore interaction over similar resources (e.g. carcasses and shelter); and the presence/absence of Early Paleolithic bone tools, art, and symbolism in Korea. PMID- 10835264 TI - Stratigraphy, artefact industries and hominid associations for Sterkfontein, member 5. AB - A revised stratigraphy for the early hominid site of Sterkfontein (Gauteng Province, South Africa) reveals a complex distribution of infills in the main excavation area between 2.8 and 1.4 m.y.a, as well as deposits dating to the mid to late Pleistocene. New research now shows that the Member 4 australopithecine breccia (2.8-2.6 Ma) extends further west than was previously thought, while a late phase of Member 4 is recognized in a southern area. The artefact-bearing breccias were defined sedimentologically as Member 5, but one supposed part of these younger breccias, the StW 53 infill, lacks in situ stone tools, although it does appear to post-date 2.6 Ma when artefacts first appear in the archaeological record. The StW 53 hominid, previously referred to Homo habilis, is here argued to be Australopithecus. The first artefact-bearing breccia of Member 5 is the Oldowan Infill, estimated at 2-1.7 Ma. It occupies a restricted distribution in Member 5 east and contains an expedient, flake-based tool industry associated with a few fossils of Paranthropos robustus. An enlarged cave opening subsequently admitted one or more Early Acheulean infills associated in Member 5 west with Homo ergaster. The artefacts attest to a larger site accumulation between ca. 1.7 and 1.4 Ma, with more intensive use of quartzite over quartz and a subtle but important shift to large flakes and heavier-duty tools. The available information on palaeoenvironments is summarized, showing an overall change from tropical to sub-tropical gallery forest, forest fringe and woodland conditions in Member 4 to more open woodland and grassland habitats in the later units, but with suggestions of a wet localized topography in the Paranthropus bearing Oldowan Infill. PMID- 10835265 TI - The age and context of the Tabun I skeleton: a reply to Schwarcz et al. PMID- 10835266 TI - A new hominid incisor from Sangiran, Central Java. PMID- 10835267 TI - Phylogenetic analysis of the cadherin superfamily allows identification of six major subfamilies besides several solitary members. AB - Cadherins play an important role in specific cell-cell adhesion events. Their expression appears to be tightly regulated during development and each tissue or cell type shows a characteristic pattern of cadherin molecules. Inappropriate regulation of their expression levels or functionality has been observed in human malignancies, in many cases leading to aggravated cancer cell invasion and metastasis. The cadherins form a superfamily with at least six subfamilies, which can be distinguished on the basis of protein domain composition, genomic structure, and phylogenetic analysis of the protein sequences. These subfamilies comprise classical or type-I cadherins, atypical or type-II cadherins, desmocollins, desmogleins, protocadherins and Flamingo cadherins. In addition, several cadherins clearly occupy isolated positions in the cadherin superfamily (cadherin-13, -15, -16, -17, Dachsous, RET, FAT, MEGF1 and most invertebrate cadherins). We suggest a different evolutionary origin of the protocadherin and Flamingo cadherin genes versus the genes encoding desmogleins, desmocollins, classical cadherins, and atypical cadherins. The present phylogenetic analysis may accelerate the functional investigation of the whole cadherin superfamily by allowing focused research of prototype cadherins within each subfamily. PMID- 10835268 TI - Large conformational changes in the maturation of a simple RNA virus, nudaurelia capensis omega virus (NomegaV). AB - An assembly intermediate of a small, non-enveloped RNA virus has been discovered that exhibits striking differences from the mature virion. Virus-like particles (VLPs) of Nudaurelia capensis omega virus (NomegaV), a T=4 icosahedral virus infecting Lepidoptera insects, were produced in insect cells using a baculovirus vector expressing the coat protein. A procapsid form was discovered when NomegaV VLPs were purified at neutral pH conditions. These VLPs were fragile and did not undergo the autoproteolytic maturation that occurs in the infectious virus. Electron cryo-microscopy (cryoEM) and image analysis showed that, compared with the native virion, the VLPs were 16% larger in diameter, more rounded, porous, and contained an additional internal domain. Upon lowering the pH to 5.0, the VLP capsids became structurally indistinguishable from the authentic virion and the subunits autoproteolyzed. The NomegaV protein subunit coordinates, which were previously determined crystallographically, were modelled into the 28 A resolution cryoEM map of the procapsid. The resulting pseudo-atomic model of the NomegaV procapsid demonstrated the large rearrangements in quaternary and tertiary structure needed for the maturation of the VLPs and presumably of the virus. Based on this model, we propose that electrostatically driven rearrangements of interior helical regions are responsible for the large conformational change. These results are surprising because large structural rearrangements have not been found in the maturation of any other small RNA viruses. However, similarities of this conformational change to the maturational processes of more complex DNA viruses (e.g. bacteriophages and herpesvirus) and to the swelling of simple plant viruses suggest that structural changes in icosahedral viruses, which are integral to their function, have similar strategies and perhaps mechanisms. PMID- 10835269 TI - Scanning force microscopy of the complexes of p53 core domain with supercoiled DNA. AB - We used scanning force microscopy to analyse the interaction of the core domain of the tumor suppressor protein p53 (p53CD, amino acid residues 94 to 312), with supercoiled DNA (scDNA) molecules. The complexes were attached to a mica substrate by the divalent cation spreading technique. p53CD bound to supercoiled plasmid pPGM1 bearing the consensus sequence 5'-AGACATGCCTAGACATGCCT-3' (p53CON) was imaged as a globular complex. Only one such complex was observed with each scDNA molecule. In contrast, binding to supercoiled pBluescript II SK(-) DNA (lacking the consensus sequence) resulted in the appearance of multiple, variable size complexes of various sizes on single DNA molecules. Addition of p53CD to scDNA containing a cruciform-forming (AT)(34) insert resulted in the binding of the protein exclusively at the cruciform. The data presented here suggest that p53CD can form stable specific and non-specific complexes with supercoiled DNA molecules, albeit of variable multimeric organization. PMID- 10835270 TI - Dimethylsulfoxide reductase: an enzyme capable of catalysis with either molybdenum or tungsten at the active site. AB - DMSO reductase (DMSOR) from Rhodobacter capsulatus, well-characterised as a molybdoenzyme, will bind tungsten. Protein crystallography has shown that tungsten in W-DMSOR is ligated by the dithiolene group of the two pyranopterins, the oxygen atom of Ser147 plus another oxygen atom, and is located in a very similar site to that of molybdenum in Mo-DMSOR. These conclusions are consistent with W L(III)-edge X-ray absorption, EPR and UV/visible spectroscopic data. W DMSOR is significantly more active than Mo-DMSOR in catalysing the reduction of DMSO but, in contrast to the latter, shows no significant ability to catalyse the oxidation of DMS. PMID- 10835271 TI - TFIIIC-independent in vitro transcription of yeast tRNA genes. AB - The most peculiar transcriptional property of eukaryotic tRNA genes, as well as of other genes served by RNA polymerase III, is their complete dependence on the intragenic interaction platform provided by transcription factor IIIC (TFIIIC) for the productive assembly of the TBP-containing initiation factor TFIIIB. The sole exception, in yeast, is the U6 RNA gene, which is able to exploit a TATAAATA element, 30 bp upstream of the transcription start site, for the TFIIIC independent assembly of TFIIIB. To find out whether this extragenic core promoter organization and autonomous TFIIIB assembly capacity are unique features of the U6 gene or also apply to other genes transcribed by RNA polymerase III, we scanned the 5'-flanking regions (up to position -100) of the entire tRNA gene set of Saccharomyces cerevisiae searching for U6-like TATA motifs. Four tRNA genes harboring such a sequence motif around position -30 were identified and found to be transcribed in vitro by a minimal system only composed of TFIIIB and RNA polymerase III. In this system, start site selection is not at all affected by the absence of TFIIIC, which, when added, significantly stimulates transcription by determining an increase in the number, rather than in the efficiency of utilization, of productive initiation complexes. A specific TBP-TATA element interaction is absolutely required for TFIIIC-independent transcription, but the nearby sequence context also contributes to the efficiency of autonomous TFIIIB assembly. The existence of a TFIIIB assembly pathway leading to the faithful transcription of natural eukaryotic tRNA genes in the absence of TFIIIC provides novel insights into the functional flexibility of the eukaryotic tRNA gene transcription machinery and on its evolution from an ancestral RNA polymerase III system relying on upstream, TATA- centered control elements. PMID- 10835272 TI - Initiation factor 3-induced structural changes in the 30 S ribosomal subunit and in complexes containing tRNA(f)(Met) and mRNA. AB - Initiation factor 3 (IF3) acts to switch the decoding preference of the small ribosomal subunit from elongator to initiator tRNA. The effects of IF3 on the 30 S ribosomal subunit and on the 30 S.mRNA. tRNA(f)(Met) complex were determined by UV-induced RNA crosslinking. Three intramolecular crosslinks in the 16 S rRNA (of the 14 that were monitored by gel electrophoresis) are affected by IF3. These are the crosslinks between C1402 and C1501 within the decoding region, between C967xC1400 joining the end loop of a helix of 16 S rRNA domain III and the decoding region, and between U793 and G1517 joining the 790 end loop of 16 S rRNA domain II and the end loop of the terminal helix. These changes occur even in the 30 S.IF3 complex, indicating they are not mediated through tRNA(f)(Met) or mRNA. UV-induced crosslinks occur between 16 S rRNA position C1400 and tRNA(f)(Met) position U34, in tRNA(f)(Met) the nucleotide adjacent to the 5' anticodon nucleotide, and between 16 S rRNA position C1397 and the mRNA at positions +9 and +10 (where A of the initiator AUG codon is +1). The presence of IF3 reduces both of these crosslinks by twofold and fourfold, respectively. The binding site for IF3 involves the 790 region, some other parts of the 16 S rRNA domain II and the terminal stem/loop region. These are located in the front bottom part of the platform structure in the 30 S subunit, a short distance from the decoding region. The changes that occur in the decoding region, even in the absence of mRNA and tRNA, may be induced by IF3 from a short distance or could be caused by the second IF3 structural domain. PMID- 10835273 TI - Radioprobing of a RecA-three-stranded DNA complex with iodine 125: evidence for recognition of homology in the major groove of the target duplex. AB - A fundamental problem in homologous recombination is how homology between DNAs is recognized. In all current models, a recombination protein loads onto a single strand of DNA and scans another duplex for homology. When homology is found, a synaptic complex is formed, leading to strand exchange and a heteroduplex. A novel technique based on strand cleavage by the Auger radiodecay of iodine 125, allows us to determine the distances between (125)I on the incoming strand and the target sugars of the duplex DNA strands in an Escherichia coli RecA protein mediated synaptic complex. Analysis of these distances shows that the complex represents a post-strand exchange intermediate in which the heteroduplex is located in the center, while the outgoing strand forms a relatively wide helix intertwined with the heteroduplex and located in its minor groove. The structure implies that homology is recognized in the major groove of the duplex. PMID- 10835274 TI - The proteasome activator 11 S REG or PA28: chimeras implicate carboxyl-terminal sequences in oligomerization and proteasome binding but not in the activation of specific proteasome catalytic subunits. AB - The REG homologs, alpha, beta and gamma, activate mammalian proteasomes in distinct ways. REGalpha and REGbeta activate the trypsin-like, chymotrypsin-like and peptidylglutamyl-preferring active sites, whereas REGgamma only activates the proteasome's trypsin-like subunit. The three REG homologs differ in carboxyl terminal sequences that are located next to activation loops on their proteasome binding surface. To assess the importance of these carboxyl-terminal sequences in the activation of specific proteasome beta catalytic subunits, we characterized chimeras in which 8 or 12 residues were exchanged among the three proteins. Like the wild-type molecule, REGalpha chimeras activated all three proteasome catalytic subunits regardless of the carboxyl-terminal sequence. However, REGalpha-beta chimeras activated the proteasome at lower concentrations than wild type REGalpha and higher levels of REGalpha-gamma chimeras were needed for maximal activation because exchanged carboxyl-terminal sequences can stabilize (REGalpha-beta) or destabilize (REGalpha-gamma) the REGalpha heptamer. REGgamma chimeras were equivalent to REGgamma in their activation properties, but they bound the proteasome less tightly than the wild-type molecule. REGbeta chimeras also bound the proteasome more weakly than wild-type REGbeta and were virtually unable to activate it. Our findings demonstrate that the carboxyl-terminal sequences of REG subunits can affect heptamer stability and proteasome affinity, but they do not determine which proteasome beta subunits become activated. PMID- 10835275 TI - Identification of the chromosomal replication origin from Thermus thermophilus and its interaction with the replication initiator DnaA. AB - The chromosomal replication origin oriC and the gene encoding the replication initiator protein DnaA from Thermus thermophilus have been identified and cloned into an Escherichia coli vector system. The replication origin is composed of 13 characteristically arranged DnaA boxes, binding sites for the DnaA protein, and an AT-rich stretch, followed by the dnaN gene. The dnaA gene is located upstream of the origin and expresses a typical DnaA protein that follows the division into four domains, as with other members of the DnaA protein family. Here, we report the purification of Thermus-DnaA (Tth-DnaA) and characterize the interaction of the purified protein with the replication origin, with regard to the binding kinetics and stoichiometry of this interaction. Using gel retardation assays, surface plasmon resonance (SPR) and electron microscopy, we show that, unlike the E. coli DnaA, Tth-DnaA does not recognize a single DnaA box, instead a cluster of three tandemly repeated DnaA boxes is the minimal requirement for specific binding. The highest binding affinities are observed with full-length oriC or six clustered, tandemly repeated DnaA boxes. Furthermore, high-affinity DNA-binding of Tth-DnaA is dependent on the presence of ATP. The Thermus DnaA/oriC interaction will be compared with oriC complex formation generated by other DnaA proteins. PMID- 10835276 TI - Selection of homeotic proteins for binding to a human DNA replication origin. AB - We have previously shown that a cell cycle-dependent nucleoprotein complex assembles in vivo on a 74 bp sequence within the human DNA replication origin associated to the Lamin B2 gene. Here, we report the identification, using a one hybrid screen in yeast, of three proteins interacting with the 74 bp sequence. All of them, namely HOXA13, HOXC10 and HOXC13, are orthologues of the Abdominal-B gene of Drosophila melanogaster and are members of the homeogene family of developmental regulators. We describe the complete open reading frame sequence of HOXC10 and HOXC13 along with the structure of the HoxC13 gene. The specificity of binding of these two proteins to the Lamin B2 origin is confirmed by both band shift and in vitro footprinting assays. In addition, the ability of HOXC10 and HOXC13 to increase the activity of a promoter containing the 74 bp sequence, as assayed by CAT-assay experiments, demonstrates a direct interaction of these homeoproteins with the origin sequence in mammalian cells. We also show that HOXC10 expression is cell-type-dependent and positively correlates with cell proliferation. PMID- 10835277 TI - The DNA damage spectrum produced by simulated sunlight. AB - The mutagenic effects of ultraviolet and solar irradiation are thought to be due to the formation of DNA photoproducts, most notably cyclobutane pyrimidine dimers (CPDs) and pyrimidine (6-4) pyrimidone photoproducts ((6-4)PPs). Experimental systems for determining the levels and sequence dependence of photoproduct formation in DNA have often used high doses of short-wave (UVC) irradiation. We have re-assessed this issue by using DNA sequencing technologies and different doses of UVC as well as more physiologically relevant doses of solar irradiation emitted from a solar UV simulator. It has been questioned whether hot alkali treatment can detect (6-4)PPs at all sequence positions. With high UVC doses, the sequence distribution of (6-4)PPs was virtually identical when hot alkali or UV damage endonuclease (UVDE) were used for detection, which appears to validate both methods. The (6-4)PPs form at 5'-TpC and 5'CpC sequences but very low levels are seen at all other dipyrimidines including 5'-TpT. Contrary to expectation, we find that (6-4) photoproducts form at almost undetectable levels under conditions of irradiation for up to five hours with the solar UV simulator. The same treatment produces high levels of CPDs. In addition, DNA glycosylases, which recognize oxidized and ring-opened bases, did not produce significant cleavage of sunlight-irradiated DNA. From these data, we conclude that cyclobutane pyrimidine dimers are at least 20 to 40 times more frequent than any other DNA photoproduct when DNA or cells are irradiated with simulated sunlight. PMID- 10835278 TI - Sequence-dependent elastic properties of DNA. AB - Harmonic elastic constants of 3-11 bp duplex DNA fragments were evaluated using four 5 ns unrestrained molecular dynamics simulation trajectories of 17 bp duplexes with explicit inclusion of solvent and counterions. All simulations were carried out with the Cornell et al. force-field and particle mesh Ewald method for long-range electrostatic interactions. The elastic constants including anisotropic bending and all coupling terms were derived by analyzing the correlations of fluctuations of structural properties along the trajectories. The following sequences have been considered: homopolymer d(ApA)(n) and d(GpG)(n), and alternating d(GPC)(n) and d(APT)(n). The calculated values of elastic constants are in very good overall agreement with experimental values for random sequences. The atomic-resolution molecular dynamics approach, however, reveals a pronounced sequence-dependence of the stretching and torsional rigidity of DNA, while sequence-dependence of the bending rigidity is smaller for the sequences considered. The earlier predicted twist-bend coupling emerged as the most important cross-term for fragments shorter than one helical turn. The calculated hydrodynamic relaxation times suggest that damping of bending motions may play a role in molecular dynamics simulations of long DNA fragments. A comparison of elasticity calculations using global and local helicoidal analyses is reported. The calculations reveal the importance of the fragment length definition. The present work shows that large-scale molecular dynamics simulations represent a unique source of data to study various aspects of DNA elasticity including its sequence-dependence. PMID- 10835280 TI - A genetic analysis of crystal growth. AB - The regulation of crystal morphology by proteins is often observed in biology. It is a central feature in the formation of hard tissues such as bones, teeth and mollusc shells. We have developed a genetic system in the bacterium Escherichia coli to study the protein-mediated control of crystal growth. We have used the crystallization of gold as a model system and found polypeptides that control the morphology of the resulting gold crystals. Analysis of the crystallization process influenced by these polypeptides indicates they act catalytically by an acid mechanism. Our results suggest that the concepts and methods of microbial genetics are general and can be applied to substances not commonly found in biological systems. PMID- 10835279 TI - Crystal structure of the 64M-2 antibody Fab fragment in complex with a DNA dT(6 4)T photoproduct formed by ultraviolet radiation. AB - DNA photoproducts with (6-4) pyrimidine-pyrimidone adducts formed by ultraviolet radiation are implicated in mutagenesis and cancer, particularly skin cancer. The crystal structure of the Fab fragment of the murine 64M-2 antibody specific to DNA T(6-4)T photoproducts is determined as a complex with dT(6-4)T, a (6-4) pyrimidine-pyrimidone photodimer of dTpT, at 2.4 A resolution to a crystallographic R-factor of 0.199 and an R(free) value of 0.279. The 64M-2 Fab molecule is in an extended arrangement with an elbow angle of 174 degrees, and its five complementarity-determining regions, except L2, are involved in the ligand binding. The bound dT(6-4)T ligand adopting a ring structure with (6-4) linked 5' thymine-3' pyrimidone bases is fully accommodated in an antigen-binding pocket of about 15 Ax10 A. The 5'-thymine and 3'-pyrimidone bases are in half chair and planar conformations, respectively, and are nearly perpendicular to each other. The 5'-thymine base is hydrogen-bonded to Arg95H and Ser96H, and is in van der Waals contact with Tyr100iH. The 3'-pyrimidone base is hydrogen-bonded to His35H, and is in contact with Trp33H. Three water molecules are located at the interface between the bases and the Fab residues. Hydrogen bonds involving these water molecules also contribute to Fab recognition of the dT(6-4)T bases. The sugar-phosphate backbone connecting the bases is surrounded by residues His27dL, Tyr32L, Ser92L, Trp33H, and Ser58H, but is not hydrogen-bonded to these residues. PMID- 10835281 TI - The structural role of the copper-coordinating and surface-exposed histidine residue in the blue copper protein azurin. AB - Copper K-edge extended X-ray absorption fine structure (EXAFS) spectroscopy and (15)N NMR relaxation studies were performed on samples of a variant azurin in which the surface-exposed histidine ligand of the copper atom (His117) has been replaced by glycine. The experiments were performed to probe the structure of the active site and the protein dynamics. The cavity in the protein structure created by the His-->Gly replacement could be filled by external ligands, which can either restore the spectroscopic properties of the original type-1 copper site or create a new type-2 copper site. The binding of external ligands occurs only when the copper atom is in its oxidised state. In the reduced form, the binding is abolished. From the EXAFS experiments, it is concluded that for the oxidised type 1 copper sites the protein plus external ligand (L) provide an NSS*L donor set deriving from His46, Cys112, Met121 and the external ligand. The type-2 copper site features an S(N/O)(3) donor set in which the S-donor derives from Cys112, one N-donor from His46 and the remaining two N or O donors from one or more external ligands. Upon reduction of the type-1 as well as the type-2 site, the external ligand drops out of the copper site and the coordination reduces to 3 fold with an SS*N donor set deriving from His46, Cys112 and Met121. The Cu S(delta)(Met) distance is reduced from about 3.2 to 2.3 A. Analysis of the NMR data shows that the hydrophobic patch around His117 has gained fluxionality when compared to wild-type azurin, which may explain why the His117Gly variant is able to accommodate a variety of external ligands of different sizes and with different chelating properties. On the other hand, the structure and dynamics of the beta-sandwich, which comprises the main body of the protein, is only slightly affected by the mutation. The unusually high reduction potential of the His117Gly azurin is discussed in light of the present results. PMID- 10835282 TI - Molten globule structure of equine beta-lactoglobulin probed by hydrogen exchange. AB - The molten globule structure of equine beta-lactoglobulin has been inferred from the hydrogen exchange protection of the backbone amide protons. In order to make it possible to measure the hydrogen exchange kinetics of the individual backbone amide protons, the uniformly (15)N-labeled recombinant protein was expressed in Escherichia coli and the NMR peak assignment was obtained for most of the backbone protons. The chemical shift and NOE results obtained under the condition where the protein assumes the native structure are fully consistent with the known secondary structure of bovine beta-lactoglobulin, indicating that the equine protein has a similar native conformation to that of the bovine protein. The hydrogen exchange rate of the individual backbone amide protons was measured under the conditions where the protein assumes the native and molten globule states. In the native state, strong protection was observed for the residues located in the eight (A to H) strands, which form a barrel structure, and residues of a major helix. In the molten globule state at acidic pH conditions, significant protection from the exchange has been observed for residues located in the A, F, G and H strands in the native structure. The pattern of protection is consistent with a native-like beta-sheet formation by these strands. The residues located in a major helix of the native structure are also protected, suggesting that this helix is formed in the molten globule and is packed against the sheet as in the native structure. These results indicate that a native-like subdomain is formed in the molten globule state of equine beta-lactoglobulin. PMID- 10835283 TI - Backbone dynamics of the C-terminal SH2 domain of the p85alpha subunit of phosphoinositide 3-kinase: effect of phosphotyrosine-peptide binding and characterization of slow conformational exchange processes. AB - The backbone dynamics of the C-terminal SH2 domain from the regulatory subunit p85alpha (p85alpha C-SH2) of phosphoinositide 3-kinase has been investigated in the absence of, and in complex with, a high-affinity phosphotyrosine-containing peptide ligand derived from the platelet-derived growth-factor receptor. (15)N R(1) and R(2) relaxation rates and steady-state [(1)H]-(15)N NOE values were measured by means of (1)H-(15)N correlated two-dimensional methods and were analyzed within the framework of the model-free formalism. Several residues in the BC loop and in the neighbouring secondary structural elements display fast local dynamics in the absence of phosphotyrosine peptide ligand as evidenced by below-average [(1)H]-(15)N NOE values. Furthermore, residue Gln41 (BC3) displays conformational exchange phenomena as indicated by an above-average R(2) relaxation rate. Upon binding of the phosphotyrosine peptide, the NOE values increase to values observed for regular secondary structure and the exchange contribution to the R(2) relaxation rate for Gln41 (BC3) vanishes. These observations indicate a loss of backbone flexibility upon ligand binding. Substantial exchange contributions for His56 (betaD4) and Cys57 (betaD5), which are known to make important interactions with the ligand, are attenuated upon ligand binding. Several residues in the betaD'-FB region and the BG loop, which contribute to the ligand binding surface of the protein, exhibit exchange terms which are reduced or vanish when the ligand is bound. Together, these observations suggest that ligand binding is accompanied by a loss of conformational flexibility on the ligand binding face of the protein. However, comparison with other SH2 domains reveals an apparent lack of consensus in the changes in dynamics induced by ligand binding. Exchange rates for individual residues were quantified in peptide-complexed p85alpha C-SH2 from the dependence of the exchange contributions on the CPMG delay in an R(2) series and show that peptide-complexed p85alpha C-SH2 is affected by multiple conformational exchange processes with exchange rate constants from 10(2) s(-1) to 7.10(3) s(-1). Mapping of the exchange-rate constants on the protein surface show a clustering of residues with similar exchange-rate constants and suggests that clustered residues are affected by a common predominant exchange process. PMID- 10835284 TI - Trading accuracy for speed: A quantitative comparison of search algorithms in protein sequence design. AB - Finding the minimum energy amino acid side-chain conformation is a fundamental problem in both homology modeling and protein design. To address this issue, numerous computational algorithms have been proposed. However, there have been few quantitative comparisons between methods and there is very little general understanding of the types of problems that are appropriate for each algorithm. Here, we study four common search techniques: Monte Carlo (MC) and Monte Carlo plus quench (MCQ); genetic algorithms (GA); self-consistent mean field (SCMF); and dead-end elimination (DEE). Both SCMF and DEE are deterministic, and if DEE converges, it is guaranteed that its solution is the global minimum energy conformation (GMEC). This provides a means to compare the accuracy of SCMF and the stochastic methods. For the side-chain placement calculations, we find that DEE rapidly converges to the GMEC in all the test cases. The other algorithms converge on significantly incorrect solutions; the average fraction of incorrect rotamers for SCMF is 0.12, GA 0.09, and MCQ 0.05. For the protein design calculations, design positions are progressively added to the side-chain placement calculation until the time required for DEE diverges sharply. As the complexity of the problem increases, the accuracy of each method is determined so that the results can be extrapolated into the region where DEE is no longer tractable. We find that both SCMF and MCQ perform reasonably well on core calculations (fraction amino acids incorrect is SCMF 0.07, MCQ 0.04), but fail considerably on the boundary (SCMF 0.28, MCQ 0.32) and surface calculations (SCMF 0.37, MCQ 0.44). PMID- 10835285 TI - Strengthening the dimerisation interface of Lac repressor increases its thermostability by 40 deg. C. AB - We increased drastically the heat stability of Lac repressor (LacR) of Escherichia coli. Wild-type tetrameric LacR denatures irreversibly at 53 degrees C. Improving hydrophobic packing at the dimerisation interface by a single substitution increases LacR heat-resistance by 40 deg. C without abolishing inducer binding at high and low temperatures. Tetrameric LacR mutants carrying substitutions of the positively charged amino acid Lys84 by each of the hydrophobic amino acids Leu, Ile and Met resist heating to temperatures up to 93 degrees C. We performed IPTG binding assays at 80 degrees C and found the mutant Lac repressors active and, thus, the core intact. Furthermore, the activity of LacR following heating is shown at room temperature by a gel retardation assay, which demonstrates normal oligomerisation state and function of the headpiece. The same mutations (K84L/I/M) in the dimer LacR331stop, carrying a stop codon in amino acid 331, increase thermostability of the dimer from 47 degrees C to 87 degrees C. LacRK84M represses beta-galactosidase activity in vivo as well as the wild-type and is sufficiently induced to allow growth on lactose. The results with both tetramer and dimer variants of LacR indicate mutual stabilisation of the tetramerisation region and the stable core. PMID- 10835287 TI - Volumetric and spectroscopic characterizations of the native and acid-induced denatured states of staphylococcal nuclease. AB - We have characterized the acid-induced denaturation of staphylococcal nuclease (SNase) at different urea concentrations by a combination of ultrasonic velocimetry, high precision densimetry, and CD spectroscopy. Our CD spectroscopic results suggest that, at low salt and acidic pH, the protein is unfolded with disrupted secondary and tertiary structures. Furthermore, as judged by far UV CD spectra, the protein is further unfolded at acidic pH upon the addition of urea up to the concentration of 1.5 M. The midpoint of the transition shifts to more neutral pH values and the cooperativity of the transition decreases as the acid induced denaturation of SNase occurs at higher urea concentrations. We find that the change in volume, Deltav, accompanying the acid-induced denaturation of SNase increases from -0.013 cm(3) g(-1) (-218 cm(3) mol(-1)) in the absence of urea to 0.011 cm(3) g(-1) (185 cm(3) mol(-1)) at 1.5 M urea. At all urea concentrations, the partial specific adiabatic compressibility, k(o)(s), of the protein decreases upon its unfolding with the values of Deltak(o)(s) equal to -6.3x10(-6) (-0.106 cm(3) mol(-1) bar(-1)), -4.5x10(-6) (-0.076 cm(3) mol(-1) bar(-1)), -4.6x10(-6) ( 0.077 cm(3) mol(-1) bar(-1)), and -3.8x10(-6) (-0.064 cm(3) mol(-1) bar(-1)) cm(3) g(-1) bar(-1) at urea concentrations of 0, 0.5, 1.0, and 1.5 M, respectively. In general, our volumetric results suggest that the acid-induced denatured state of SNase is only partially unfolded with the solvent-exposed surface area equal to 70-80 % of that expected for the fully extended conformation. PMID- 10835286 TI - Mg(2+) binding to tRNA revisited: the nonlinear Poisson-Boltzmann model. AB - Our current understanding of Mg(2+) binding to RNA, in both thermodynamic and structural terms, is largely based on classical studies of transfer RNAs. Based on these studies, it is clear that magnesium ions are crucial for stabilizing the folded structure of tRNA. We present here a rigorous theoretical model based on the nonlinear Poisson-Boltzmann (NLPB) equation for understanding Mg(2+) binding to yeast tRNA(Phe). We use this model to interpret a variety of experimental Mg(2+) binding data. In particular, we find that the NLPB equation provides a remarkably accurate description of both the overall stoichiometry and the free energy of Mg(2+) binding to yeast tRNA(Phe) without any fitted parameters. In addition, the model accurately describes the interaction of Mg(2+) with localized regions of the RNA as determined by the pK(a) shift of differently bound fluorophores. In each case, we find that the model also reproduces the univalent salt-dependence and the anticooperativity of Mg(2+) binding. Our results lead us to a thermodynamic description of Mg(2+) binding to yeast tRNA(Phe) based on the NLPB equation. In this model, Mg(2+) binding is simply explained by an ensemble of ions distributed according to a Boltzmann weighted average of the mean electrostatic potential around the RNA. It appears that the entire ensemble of electrostatically bound ions superficially mimics a few strongly coordinated ions. In this regard, we find that Mg(2+) stabilizes the tertiary structure of yeast tRNA(Phe) in part by accumulating in regions of high negative electrostatic potential. These regions of Mg(2+) localization correspond to bound ions that are observed in the X-ray crystallographic structures of yeast tRNA(Phe). Based on our results and the available thermodynamic data, there is no evidence that specifically coordinated Mg ions have a significant role in stabilizing the native tertiary structure of yeast tRNA(Phe) in solution. PMID- 10835289 TI - Nitric oxide-mediated erectile effects of galantide but not galanin in vivo. AB - The purpose of this study was to investigate the in vivo effects of intracavernosal injections of galanin and galantide (a specific galanin receptor antagonist) on penile erection in the anesthetized cat. Erectile responses to galanin and galantide were compared with responses to a standard triple drug combination [1.65 mg papaverine, 25 microg phentolamine, and 0.5 microg prostaglandin E(1) (PGE(1))]. Intracavernosal injections of galanin (3-100 nmol) and galantide (0. 1-3 nmol) induced penile erection in a dose-dependent manner. In terms of relative potency, galantide was approximately 100-fold more potent than galanin at increasing cavernosal pressure. The maximal increases in intracavernosal pressure in response to galanin and galantide were 83 and 95%, respectively, of the control triple drug combination. The total durations of erectile response caused by these peptides were significantly shorter (P<0.05) than those by the triple drug combination. The nitric oxide synthase inhibitor L NAME (20 mg) significantly decreased the erectile response in the cat to galantide but not to galanin, while the K(+)(ATP) channel antagonist U-37883A (3 mg) had no effect on the erectile response to galanin nor galantide. The results of the present study demonstrate that galantide, a putative antagonist for the galanin receptor, has more potent agonist activity than galanin in increasing intracavernosal pressure in the cat. Moreover, these data suggest that galantide, but not galanin, causes penile erection by an NO/cGMP-dependent mechanism. This is the first study to demonstrate that galanin may play a role in the physiology of penile erection. PMID- 10835288 TI - Duration of the hemodynamic effects of N(G)-nitro-L-arginine methyl ester in vivo. AB - The objective of this study was to quantify the duration of the hemodynamic activity of N(G)-nitro-l-arginine methyl ester (l-NAME) in a variety of different tissues following a single bolus injection of this nitric oxide synthase inhibitor to healthy rats. l-NAME (15 micromol x kg(-1)) was injected (ip) into rats to produce maximal inhibition of endothelial cell NOS. Animals were subsequently anesthetized and blood flow was quantified using the radioactive microsphere/reference organ technique. At 1 h following a single bolus injection of l-NAME blood flow was reduced to the entire gastrointestinal tract, pancreas, and liver. Three hours following l-NAME administration, blood flow to the stomach and upper small intestine had returned to pretreatment levels; however, blood flow to the jejunum, ileal-jejunal junction, and colon remained significantly reduced. Splenic blood flow was significantly reduced and hepatic arterial blood flow was further reduced at this time as well. After 6 h following l-NAME administration, blood flow in all organs had completely recovered to control levels. Although cardiac index and total peripheral resistance had also returned to preinjection values at this time, mean arterial pressure remained elevated at 6 h posttreatment. Blood flow to the brain, lungs, and psoas muscle were unaffected by l-NAME administration at any time point. Taken together, these data demonstrate a differential regulation of vascular tone by NO in different vascular beds and, depending upon the organ system in question, the vasoactive activity of l-NAME may last from 3 to 6 h following a single bolus injection of this NOS inhibitor. PMID- 10835290 TI - Phagocytosis is regulated by nitric oxide in murine microglia. AB - Nitric oxide (NO) is produced by inducible nitric oxide synthase (iNOS) in activated microglia and has been shown to participate in host defense mechanisms. However, the role of NO produced by constitutive nitric oxide synthase (cNOS) in microglia is poorly understood. In this report, NO was found to regulate phagocytosis in murine BV-2 microglial cells as quantified by flow cytometry. Addition of NO-generating compounds caused impaired phagocytosis as compared to untreated microglia. The addition of nitric oxide synthase (NOS) inhibitors to microglial cells resulted in potentiation of phagocytosis, suggesting that constitutive NO was participating in the regulation of phagocytosis. The inverse correlation between NO production and phagocytosis was also observed when Alzheimer's beta-amyloid peptide was added. With beta-amyloid treatment, constitutive NO production decreased while phagocytosis increased. Cell extracts prepared from untreated microglia were found to contain both neuronal and endothelial NOS isoforms, but not the inducible form. The correlation of spontaneous NO production with attenuated phagocytosis suggests that constitutive NOS enzymes participate in microglial regulation. PMID- 10835291 TI - Effects of short-term nitrogen monoxide inhalation on leukocyte adhesion molecules, generation of reactive oxygen species, and cytokine release in human blood. AB - Increased nitrogen monoxide (NO) concentrations change leukocyte function under a multitude of experimental conditions. NO inhalation is an experimental treatment for lung failure and exposes leukocytes to increased NO concentrations during passage through the lungs. To investigate whether short-term NO inhalation induces lasting changes in the function of circulating human leukocytes, venous blood samples were drawn from eight healthy male volunteers before and at the end of a 35-min period of breathing 40 ppm NO in 30% O(2). The leukocytes in the samples were subsequently analyzed for NO-induced changes in expression of cell surface molecules, generation of reactive oxygen species (ROS), and cytokine production by flow cytometry and ELISA techniques. The results were (1) NO inhalation changed neither the baseline nor the Escherichia coli lipopolysaccharide (LPS)-induced expression of the cell adhesion molecules CD11a, CD11b, CD11c, and CD62L (l-selectin) on neutrophilic granulocytes (PMN) or monocytes (Mo). The expression of CD14 and HLA-DR was also unchanged. (2) The generation of ROS in response to activation with phorbol myristate acetate increased in PMN after NO inhalation; an increase in Mo did not reach significance. (3) Baseline and LPS-stimulated production of IL-1beta decreased after NO inhalation, while the LPS-stimulated production of TNF-alpha increased. No changes in IL-6 production were detected. PMID- 10835292 TI - Pulmonary blast injury increases nitric oxide production, disturbs arginine metabolism, and alters the plasma free amino acid pool in rabbits during the early posttraumatic period. AB - Plasma nitrate + nitrite (nitrates), as final NO products, and free amino acid pool (FAAP) characteristics, as indicators of protein/amino acid metabolism, were analyzed in the early (30 min) period following blast injury. The experiments were performed on 27 rabbits subjected to pulmonary blast injury (experimental group) or not exposed to overpressure (controls). We report that pulmonary blast injury (PBI) induces prompt NO overproduction within a very early period. Increased arginine utilization via NO synthase, presumably associated with its cleavage by arginase, leads to the depletion of the arginine level in arterial plasma 30 min following PBI. Impaired balance between arginine utilization and release/resynthesis from endogenous sources causes disturbed nutritional status and urea cycle activity. Early identification and appropriate management of the changes in amino acid metabolism should be included in the evaluation of patients with blast injury. Furthermore, the results suggest that depleted arterial levels of arginine and NO overproduction may be helpful in diagnosis and prognosis of blast injury. PMID- 10835293 TI - Involvement of nitric oxide in nickel-induced hyperglycemia in rats. AB - Nitric oxide is an important bioactive signaling molecule that mediates a variety of normal physiological functions which, if altered, could contribute to the genesis of many pathological conditions, including diabetes. In the present study we have shown the involvement of NO in nickel-induced hyperglycemia in male albino rats. Administration of nickel chloride (25 to 100 micromol/kg; ip) to overnight-fasted rats resulted in significant dose and time-dependent increase in plasma glucose, attaining maximum level at 1 h posttreatment and thereafter decreasing to normal levels by 4 h. The involvement of NO in nickel-induced hyperglycemia was evident by the observation that pretreatment of rats with NG monomethyl-l-arginine (10 to 50 micromol/kg; ip), an inhibitor of nitric oxide synthase (NOS), significantly attenuated the nickel-mediated increase in the plasma glucose levels in a dose-dependent fashion. The activity of Ca(2+) dependent NOS (constitutive form, c-NOS) was found to be significantly elevated in adrenals (5.5-fold) and brain (1.4-fold) at 1 and 2 h posttreatment, attaining normal levels by 4 h. In contrast, the activity of c-NOS in pancreas was significantly decreased (2.8-fold) with a concomitant increase (11.6-fold) in inducible NOS (i-NOS) at the same time interval. As observed by immunoblot analysis, a significant increase in i-NOS protein expression in the pancreas was observed at 1 and 2 h posttreatment. This was associated with a significant elevation in cGMP levels in adrenals, brain, and pancreas, possibly via the stimulation of cytosolic guanylate cyclase. This elevation in cGMP was abolished by low concentration of hemoglobin. These effects were associated with the accumulation of nickel in the target tissues. Taken together, our data suggest that nickel causes a significant increase in the levels of (i) cGMP and c-NOS in adrenals and brain and (ii) i-NOS in pancreas. These events may be responsible for modulating the release of insulin from pancreas finally leading to hyperglycemic condition in rats. PMID- 10835294 TI - Blood pressure changes after intravenous administration of cell-free hemoglobin A and hemoglobin H in the rat. AB - Hemoglobin H (HbH) is a tetramer of four beta chains (present in erythrocytes of alpha thalassemia), whereas hemoglobin A is a tetramer of two alpha and two beta chains. Since HbH is known to bind four times more nitric oxide (a vasodilator) at its sulfhydryls compared to HbA, the present studies were conducted to see the effect of HbH and HbA on rat blood pressure. The acute administration (20-2000 nmol/kg) of both HbH and HbA produced a dose-dependent effect on blood pressure. The net change in mean arterial pressure was significantly higher with HbH compared to HbA. Partially nitrosylated (in which SH groups are occupied with NO) HbH retained the property of raising blood pressure to some extent while HbA lost this property. Completely nitrosylated (in which both heme and SH groups are occupied with NO) derivatives of both HbH and HbA reduced the blood pressure to the same extent. The preliminary studies with chronic administration of HbA and HbH resulted in nonsignificant increase in blood pressure. It is concluded that these findings may explain the earlier observations of increased risk of hypertension in individuals with alpha thalassemia. PMID- 10835295 TI - Transformations of 2,6-diisopropylphenol by NO-derived nitrogen oxides, particularly peroxynitrite. AB - To investigate the protective effect of the anesthetic 2, 6-diisopropylphenol, or propofol, in oxidative processes in which (*)NO and peroxynitrite are involved, direct interactions were explored. The reactions of the highly lipophilic propofol with (*)NO in methanolic or aqueous buffered solutions under air were shown to produce the same compounds as those detected with peroxynitrite, but with very low yields and slow rates. In aqueous neutral medium, peroxynitrite (ONOO(-), ONOOCO(-)(2), ONOOH) was able to nitrate and oxidize propofol: In addition to oxidation products, quinone and quinone dimer, the formation of the 4 nitropropofol derivative was detected, increasing with peroxynitrite or CO(2) concentrations. Nitration reached 20% after the addition of 25 mM bicarbonate to an equimolecular mixture of peroxynitrite and propofol in methanol/phosphate buffered solution (1/4,v/v) at pH 7.4. However, peroxynitrite either in methanol or in alkaline-buffered mixture (optimum pH 10-12) resulted in the rapid and almost complete transformation of propofol to an intermediate compound 1, which further decomposed to 4-nitrosopropofol. The transient compound 1 was obtained from either peroxynitrite or (*)NO in the presence of oxygen. From mass spectrometry determination of compound 1 we propose the involvement of the nitrosodioxyl radical ONOO(*), forming an adduct with the propofoxyl radical, to yield 4-nitrosodioxypropofol and finally 4-nitrosopropofol. PMID- 10835296 TI - Inhibition of the cytokine-mediated inducible nitric oxide synthase expression in rat insulinoma cells by phenyl N-tert-butylnitrone. AB - Cytokines and nitric oxide (NO) have been implicated in the pathogenesis of insulin-dependent diabetes mellitus (IDDM). We have shown that the spin-trapping agent phenyl N-tert-butylnitrone (PBN) protects against streptozotocin (STZ) induced IDDM in mice. In order to gain more insights into the mechanism(s) of the protective action of PBN against IDDM, we have investigated the effect of this compound on the cytokine-induced NO generation (measured as nitrite) in rat insulinoma RIN-5F cells. Our results demonstrate that PBN cotreatment prevents the generation of nitrite by RIN-5F cells induced by treatment with tumor necrosis factor-alpha, interleukin 1beta, and interferon-gamma in a dose dependent fashion. The generation of NO as a result of cytokine treatment and the inhibitory effect of PBN were further confirmed by electron paramagnetic resonance spectroscopy. Aminoguanidine, a selective inhibitor of inducible nitric oxide synthase (iNOS), abolished the cytokine-induced nitrite generation whereas N-nitro-l-arginine, an inhibitor more selective for other NOS isoforms, was significantly less effective. Western and Northern analyses demonstrated that PBN inhibits the cytokine-mediated expression of iNOS at the transcriptional level. Cytokine-induced nitrite formation was also inhibited by the two antioxidant agents alpha-lipoic acid and N-acetylcysteine. These results indicate that PBN protects against IDDM at least in part by prevention of cytokine-induced NO generation by pancreatic beta-cells. PMID- 10835297 TI - A simple method to sample exhaled NO not contaminated by ambient NO from children and adults in epidemiological studies. AB - We previously showed that contamination of exhaled air by ambient NO could be avoided by 1 min of breathing and final inhalation of clean air (clean air procedure) prior to exhaled air sampling in balloons. This approach is, however, unsuitable for sampling large groups in epidemiological studies, because it is time consuming and laborious. We therefore discarded the initial part of exhaled air, which may contain ambient NO, in prebags of 250, 540, 775, 1000, and 2000 ml. The subsequent part of exhaled air was sampled in balloons and the NO content was measured. Inflation of a prebag of 500 ml to prevent ambient NO contamination proved to be effective only at low ambient NO levels (<20 ppb). Larger sizes of the prebag (1000 ml for adults and 775 ml for children) are, however, required so that contamination of the air sample at higher levels of ambient NO (up to 115 ppb) is excluded. Using different prebags of gradually increasing size, it was shown that the initial part of exhaled air (<500 ml) contained relatively high amounts of NO that gradually decreased, but attained a constant level in the subsequent air volumes. Using rather large prebags of 2000 and 1000 ml, respectively, in adults and children yielded exhaled NO levels even below those obtained the clean air procedure was applied in combination with a prebag of 540 ml. As this reduction also occurs at ambient NO levels of nearly zero, we suggest that this reduction was due to interference by the water vapor arising from the lowest part of the lungs. In conclusion, the use of a prebag to discard the initial volume of exhaled air ensures accurate measurement of exhaled endogenous NO in large-scale epidemiological studies not biased by ambient NO. PMID- 10835298 TI - False nitric oxide synthase immunoreactivity in Asparagus bean (Vigna sesquipdalis) PMID- 10835300 TI - On the Role of Minor Galaxy Mergers in the Formation of Active Galactic Nuclei. AB - The large-scale ( approximately 100 kpc) environments of Seyfert galaxies are not significantly different from those of non-Seyfert galaxies. In the context of the interaction model of the formation of active galactic nuclei (AGNs), it has thus been proposed that AGNs form via "minor mergers" of large disk galaxies with smaller companions. We test this hypothesis by comparing the nuclear spectra of 105 bright nearby galaxies with measurements of their R- or r-band morphological asymmetries at three successive radii. We find no significant differences between these asymmetries among the 13 Seyfert galaxies in the sample and galaxies having other nuclear spectral types (absorption, H ii region-like, LINER), nor is there strong qualitative evidence that such mergers have occurred among any of the Seyfert galaxies or LINERs. Thus, either any minor mergers began greater, similar1 Gyr ago and are essentially complete, or they did not occur at all, and AGNs form independently of any type of interaction. Support for the latter interpretation is provided by the growing evidence that supermassive black holes exist in the cores of most elliptical and early-type spiral galaxies, which in turn suggests that nuclear activity represents a normal phase in the evolution of the bulges of massive galaxies. Galaxy mergers may increase the luminosity of Seyfert nuclei to the level of QSOs, which could explain why the latter objects appear to be found in rich environments and in interacting systems. PMID- 10835299 TI - Bayesian fine-scale mapping of disease loci, by hidden Markov models. AB - We present a new multilocus method for the fine-scale mapping of genes contributing to human diseases. The method is designed for use with multiple biallelic markers-in particular, single-nucleotide polymorphisms for which high density genetic maps will soon be available. We model disease-marker association in a candidate region via a hidden Markov process and allow for correlation between linked marker loci. Using Markov-chain-Monte Carlo simulation methods, we obtain posterior distributions of model parameter estimates including disease gene location and the age of the disease-predisposing mutation. In addition, we allow for heterogeneity in recombination rates, across the candidate region, to account for recombination hot and cold spots. We also obtain, for the ancestral marker haplotype, a posterior distribution that is unique to our method and that, unlike maximum-likelihood estimation, can properly account for uncertainty. We apply the method to data for cystic fibrosis and Huntington disease, for which mutations in disease genes have already been identified. The new method performs well compared with existing multi-locus mapping methods. PMID- 10835301 TI - The Effect of a Nonthermal Tail on the Sunyaev-Zeldovich Effect in Clusters of Galaxies. AB - We study the spectral distortions of the cosmic microwave background radiation induced by the effect in clusters of galaxies when the target electrons have a modified Maxwell-Boltzmann distribution with a high-energy nonthermal tail. Bremsstrahlung radiation from this type of electron distribution may explain the suprathermal X-ray emission observed in some clusters such as the Coma Cluster and A2199 and serve as an alternative to the classical but problematic inverse Compton scattering interpretation. We show that the Sunyaev-Zeldovich effect can be used as a powerful tool to probe the electron distribution in clusters of galaxies and discriminate among these different interpretations of the X-ray excess. The existence of a nonthermal tail can have important consequences for cluster-based estimators of cosmological parameters. PMID- 10835302 TI - Local Group Dwarf Galaxies and the Star Formation Law at High Redshift. AB - I show how the existing observational data on Local Group dwarf galaxies can be used to estimate the average star formation law during the first 3 Gyr of the history of the universe. I find that the observational data are consistent with the orthodox Schmidt law with a star formation efficiency of about 4% if the star formation is continuous (during the first 3 Gyr). The efficiency is proportionally higher if most of the gas in the dwarfs was consumed (and never replenished) in a short time interval well before the universe turned 3 Gyr. PMID- 10835303 TI - Iron K Line Variability in the Low-Luminosity Active Galactic Nucleus NGC 4579. AB - We present results of new ASCA observations of the low-luminosity active galactic nucleus (LLAGN) NGC 4579 obtained on 1998 December 18 and 28, and we report on the detection of variability of an iron K emission line. The X-ray luminosities in the 2-10 keV band for the two observations are nearly identical (LX approximately 2x1041 ergs s(-1)), but they are approximately 35% larger than that measured in 1995 July by Terashima et al. An Fe K emission line is detected at 6.39+/-0.09 keV (source rest frame), which is lower than the line energy 6.73+0.13-0.12 keV in the 1995 observation. If we fit the Fe lines with a blend of two Gaussians centered at 6.39 and 6.73 keV, the intensity of the 6.7 keV line decreases, while the intensity of the 6.4 keV line increases, within an interval of 3.5 yr. This variability rules out thermal plasmas in the host galaxy as the origin of the ionized Fe line in this LLAGN. The detection and variability of the 6.4 keV line indicates that cold matter subtends a large solid angle viewed from the nucleus and that it is located within approximately 1 pc from the nucleus. It could be identified with an optically thick standard accretion disk. If this is the case, a standard accretion disk is present at the Eddington ratio of Lbol/LEdd approximately 2x10-3. A broad disk-line profile is not clearly seen, and the structure of the innermost part of accretion disk remains unclear. PMID- 10835304 TI - Deficiency of "Thin" Stellar Bars in Seyfert Host Galaxies. AB - Using all available major samples of Seyfert galaxies and their corresponding closely matched control samples of nonactive galaxies, we find that the bar ellipticities (or axial ratios) in Seyfert galaxies are systematically different from those in nonactive galaxies. Overall, there is a deficiency of bars with large ellipticities (i.e., "thin" or "strong" bars) in Seyfert galaxies compared to nonactive galaxies. Accompanied with a large dispersion due to small number statistics, this effect is strictly speaking at the 2 sigma level. To obtain this result, the active galaxy samples of near-infrared surface photometry were matched to those of normal galaxies in type, host galaxy ellipticity, absolute magnitude, and, to some extent, redshift. We discuss possible theoretical explanations of this phenomenon within the framework of galactic evolution, and, in particular, of radial gas redistribution in barred galaxies. Our conclusions provide further evidence that Seyfert hosts differ systematically from their nonactive counterparts on scales of a few kiloparsecs. PMID- 10835305 TI - Structure of the Circumnuclear Region of Seyfert 2 Galaxies Revealed by Rossi X Ray Timing Explorer Hard X-Ray Observations of NGC 4945. AB - NGC 4945 is one of the brightest Seyfert galaxies on the sky at 100 keV, but is completely absorbed below 10 keV; its absorption column is probably the largest that still allows a direct view of the nucleus at hard X-ray energies. Our observations of it with the Rossi X-Ray Timing Explorer (RXTE) satellite confirm the large absorption, which for a simple phenomenological fit using an absorber with solar abundances implies a column of 4.5+0.4-0.4x1024 cm(-2). Using a more realistic scenario (requiring Monte Carlo modeling of the scattering), we infer the optical depth to Thomson scattering of approximately 2.4. If such a scattering medium were to subtend a large solid angle from the nucleus, it should smear out any intrinsic hard X-ray variability on timescales shorter than the light-travel time through it. The rapid (with a timescale of approximately 1 day) hard X-ray variability of NGC 4945 discovered by us with RXTE implies that the bulk of the extreme absorption in this object does not originate in a parsec size, geometrically thick molecular torus. Instead, the optically thick material on parsec scales must be rather geometrically thin, subtending a half-angle less than 10 degrees, and it is likely to be the same disk of material that is responsible for the water maser emission observed in NGC 4945. Local number counts of Seyfert 1 and Seyfert 2 galaxies show a large population of heavily obscured active galactic nuclei (AGNs) which are proposed to make up the cosmic X ray background (CXRB). However, for this to be the case, the absorption geometry in the context of axially symmetric unification schemes must have the obscuring material subtending a large scale height-contrary to our inferences about NGC 4945-implying that NGC 4945 is not a prototype of obscured AGNs postulated to make up the CXRB. The small solid angle of the absorber, together with the black hole mass (of approximately 1.4x106 M( middle dot in circle)) from megamaser measurements, allows a robust determination of the nuclear luminosity, which in turn implies that the source radiates at approximately 10% of the Eddington limit. PMID- 10835306 TI - Bar Dissolution in Prolate Halos. AB - The time evolution of barred structures is examined under the influence of the external forces exerted by a spherical halo and by prolate halos. In particular, galaxy disks are placed in the plane including the major axis of prolate halos, whose configuration is often found in cosmological simulations. N-body disks in fixed external halo fields are simulated, so that bars are formed via dynamical instability. In the subsequent evolution, the bars in prolate halos dissolve gradually with time, while the bar pattern in a spherical halo remains almost unchanged to the end of the simulation. The decay times of the bars suggest that they can be destroyed in a time smaller than a Hubble time. Our results indicate that this dissolution process could occur in real barred galaxies, if they are surrounded by massive dark prolate halos, and the configuration adopted here is not unusual from the viewpoint of galaxy formation. For a prolate halo model, an additional simulation that is restricted to two-dimensional in-plane motions has also ended up with similar bar dissolution. This means that the vertical motions of disk stars do not play an essential role in the bar dissolution demonstrated here. PMID- 10835307 TI - Discovery of a Subsystem of Globular Clusters in M31: Possible Evidence for a Past Merging Event. AB - We found a subsystem of globular clusters in M31 that appears to have a formation history separate from the rest. The member clusters of this subsystem, identified from their distinct distribution in the metallicity-distance diagram, (1) have a common medium metallicity range of -0.9<&sqbl0;Fe&solm0;H&sqbr0;<-0.4, (2) have a line-of-sight velocity distribution with no significant projected rotational motion, and (3) are populated outside the inner subsystem of globular clusters of similar metallicity that shows a significant net rotational motion as a whole. We suggest that this subsystem of globular clusters was formed during a merging event of a dwarf galaxy that took place well after the initial collapse event when most of the globular clusters in M31 were formed. PMID- 10835308 TI - Hubble Space Telescope/NICMOS Observations of I Zw 18: A Population of Old Asymptotic Giant Branch Stars Revealed. AB - I present the first results from a Hubble Space Telescope/NICMOS imaging study of the most metal-poor blue compact dwarf galaxy, I Zw 18. The near-infrared color magnitude diagram (CMD) is dominated by two populations, one 10-20 Myr population of red supergiants and one 0.1-5 Gyr population of asymptotic giant branch stars. Stars older than 1 Gyr are required to explain the observed CMD at the adopted distance of 12.6 Mpc, showing that I Zw 18 is not a young galaxy. The results hold also if the distance to I Zw 18 is significantly larger. This rules out the possibility that I Zw 18 is a truly young galaxy formed recently in the local universe. PMID- 10835309 TI - Collisional Dark Matter and the Structure of Dark Halos. AB - We study how the internal structure of dark halos is affected if cold dark matter particles are assumed to have a large cross section for elastic collisions. We identify a cluster halo in a large cosmological N-body simulation and resimulate its formation with progressively increasing resolution. We compare the structure found in the two cases in which dark matter is treated as collisionless or as a fluid. For the collisionless case, the overall ellipticity of the cluster, the central density cusp, and the amount of surviving substructure are all similar to those found in earlier high-resolution simulations. Collisional dark matter results in a cluster that is more nearly spherical at all radii, has a steeper central density cusp, and has less-but still substantial-surviving substructure. As in the collisionless case, these results for a "fluid" cluster halo are expected to carry over approximately to smaller mass systems. The observed rotation curves of dwarf galaxies then argue that self-interacting dark matter can only be viable if intermediate cross sections produce structure that does not lie between the extremes we have simulated. PMID- 10835310 TI - Age Difference between the Populations of Binary and Single F Stars Revealed from Hipparcos Data. AB - We have compared the kinematics and metallicity of the main-sequence binary and single uvby F stars from the Hipparcos catalog to see if the populations of these stars originate from the same statistical ensemble. The velocity dispersions of the known unresolved binary F stars have been found to be dramatically smaller than those of the single F stars. This suggests that the population of these binaries is, in fact, younger than that of the single stars, which is further supported by the difference in metal abundance: the binaries turn out to be, on average, more metal rich than the single stars. So, we conclude that the population of these binaries is indeed younger than that of the single F stars. Comparison of the single F stars with the C binaries (binary candidates identified in Suchkov & McMaster) has shown, on the other hand, that the latter stars are, on average, older than the single F stars. We suggest that the age difference between the single F stars, known unresolved binaries, and C binaries is associated with the fact that stellar evolution in a binary systems depends on the binary components' mass ratio and separation, with these parameters being statistically very different for the known binaries and C binaries (e.g., mostly substellar secondaries in C binaries vs. stellar secondaries in known binaries). In general we conclude that the populations of known binaries, C binaries, and single F stars do not belong to the same statistical ensemble. The implications of the discovered age difference between these populations along with the corresponding differences in kinematics and metallicity should be important not only for understanding the evolution of stars but also for the history of star formation and the evolution of the local Galactic disk. PMID- 10835311 TI - Detection of Interstellar CH3. AB - Observations with the Short Wavelength Spectrometer on board the Infrared Space Observatory have led to the first detection of the methyl radical CH(3) in the interstellar medium. The nu(2) Q-branch at 16.5 um and the R(0) line at 16.0 um have been unambiguously detected toward the Galactic center Sagittarius A*. The analysis of the measured bands gives a column density of &parl0;8.0+/ 2.4&parr0;x1014 cm(-2) and an excitation temperature of 17+/-2 K. Gaseous CO at a similarly low excitation temperature and C(2)H(2) are detected for the same line of sight. Using constraints on the H(2) column density obtained from C(18)O and visual extinction, the inferred CH(3) abundance is &parl0;1.3+2.2-0.7&parr0;x10 8. The chemically related CH(4) molecule is not detected, but the pure rotational lines of CH are seen with the Long Wavelength Spectrometer. The absolute abundances and the CH(3)/CH(4) and CH(3)/CH ratios are inconsistent with published pure gas-phase models of dense clouds. The data require a mix of diffuse and translucent clouds with different densities and extinctions, and/or the development of translucent models in which gas-grain chemistry, freeze-out, and reactions of H with polycyclic aromatic hydrocarbons and solid aliphatic material are included. PMID- 10835312 TI - On 7Li Enrichment by Low-Mass Metal-Poor Red Giant Branch Stars. AB - First-ascent red giants with strong and very strong Li lines have just been discovered in globular clusters. Using the stellar internal prompt (7)Li enrichment-mass-loss scenario, we explore the possibility of (7)Li enrichment in the interstellar matter of the globular cluster M3 produced by these Li-rich giants. We found that enrichment as large as 70% or more compared to the initial (7)Li content of M3 can be obtained during the entire life of this cluster. However, because M3 will cross into the Galactic plane several times, the new (7)Li will be very probably removed by ram pressure into the disk. Globular clusters appear then as possible new sources of (7)Li in the Galactic disk. It is also suggested that the known Na/Al variations in stars of globular clusters could be somehow related to the (7)Li variations and that the cool bottom process mixing mechanism acting in the case of (7)Li could also play a role in the case of Na and Al surface enrichments. PMID- 10835313 TI - Relativistic Effects in the Pulse Profile of the 2.5 Millisecond X-Ray Pulsar SAX J1808.4-3658. AB - I analyze the properties of the pulsed emission from the accreting millisecond pulsar SAX J1808.4-3658 in observations of its 1998 April outburst by the Rossi X Ray Timing Explorer. Pulse phase spectroscopy shows that the emission evolves from a hard spectrum (power law with photon index of 2.39+/-0.06) to a soft spectrum (index of 3.39+/-0.24). This softening is also observable as a phase lag in the fundamental of low-energy photons with respect to high-energy photons. I show that this lag is roughly constant over 10 days of the outburst. I fit these data with a model in which the pulse emission is from a hot spot on the rotating neutron star and the flux as a function of phase is determined in a calculation which includes the effects of general relativity. The energy-dependent lags are very well described by this model. The harder spectra at earlier phases (i.e., as the spot approaches) are the result of larger Doppler-boosting factors that are important for this fast pulsar. Since this model is sensitive to the equatorial speed as an independent parameter and since the spin frequency is known, this offers us a new means of measuring the neutron star radius, which is notoriously difficult to determine. PMID- 10835314 TI - Discovery of High-Frequency Quasi-periodic Oscillations in the Black Hole Candidate XTE J1859+226. AB - We report the discovery of quasi-periodic oscillations (QPOs) at roughly 187 and 150 Hz in the X-ray intensity of X-ray nova XTE J1859+226. The source was observed during a recent outburst with the Rossi X-Ray Timing Explorer. Besides these high-frequency QPOs, we have also detected QPOs (and sometimes their harmonics) at 6-7 Hz and significant broadband variability at low frequencies. These properties, as well as the observed hard X-ray spectrum, make XTE J1859+226 a black hole candidate (BHC). The detection of QPOs at two distinct frequencies greater, similar100 Hz in two observations separated by about 4 hr provide additional insights into the high-frequency QPO phenomenon in BHCs. The importance lies in the proposed interpretations, which invariably involve the effects of strong gravity near a black hole. We compare our results to those of other BHCs and discuss the impact of the observational data on the models in a global context. PMID- 10835315 TI - Protein kinase C-alpha mediates endothelial barrier dysfunction induced by TNF alpha. AB - We tested the hypothesis that protein kinase C-alpha (PKC-alpha) mediates tumor necrosis factor-alpha (TNF-alpha)-induced alterations in permeability of pulmonary microvessel endothelial monolayers (PEM). The permeability of PEM was assessed by the clearance rate of Evans blue-labeled albumin. PEM lysates were analyzed for PKC-alpha mRNA (Northern cDNA blot), protein (Western immunoblot), and activity (translocation and phosphorylation of myristoylated arginine-rich C kinase substrate). Incubation of PEM with TNF-alpha (1,000 U/ml) for 4 h resulted in increases in 1) PKC-alpha protein, 2) cytoskeletal-associated PKC-alpha, 3) PKC-alpha activity, and 4) permeability to albumin. The TNF-alpha-induced increase in PKC-alpha protein, PKC-alpha activity, and permeability was prevented by a 4-h pretreatment with PKC-alpha antisense oligonucleotide but not by the scrambled nonsense oligonucleotide. The TNF-alpha-induced increase in permeability to albumin was prevented by myristoylated protein kinase C inhibitor (an inhibitor of PKC-alpha/beta, 100 microM) and calphostin (an inhibitor of the classic and novel PKC isotypes, 200 nM). The treatment with calphostin from 0.5 to 3.0 h after TNF-alpha still prevented barrier dysfunction induced by 4 h of TNF-alpha treatment. The data indicate that prolonged activation of PKC-alpha, maintained by a translation-dependent pool of PKC-alpha protein, mediates TNF alpha-induced increases in endothelial permeability in PEM. PMID- 10835316 TI - Characterization of mucins from cultured normal human tracheobronchial epithelial cells. AB - Early-passage normal human tracheobronchial epithelial (NHTBE) cells grown in air liquid interface cultures in medium containing retinoids differentiate into a mucociliary epithelium over a 2- to 3-wk period and express increasing mRNA levels of the airway mucin genes MUC5AC and MUC5B as the cultures age; the levels of MUC2 mRNA were very low throughout the study. Using specific antibodies to MUC5AC and MUC5B mucins, we noted a gradual increase in these two mucins in the intracellular and apically secreted pools as a function of time. A low level of MUC2 mucin was detected, which did not change with time. The intracellular and apically secreted mucins isolated from day 14 and day 21 cultures by density gradient centrifugation were similar in density to those previously isolated from human respiratory mucus secretions. The sedimentation rate of the apically secreted mucins indicated that they were highly oligomerized, polydisperse macromolecules similar to those previously documented from in vivo secretions. In contrast, the cell-associated mucins from the cultured NHTBE cells were much smaller, possibly only monomers and dimers. Anion-exchange chromatography detected no differences in charge density between the reduced and carboxymethylated cell-associated and secreted forms of the MUC5AC and MUC5B mucins. The MUC5AC mucin was of similar charge density to its in vivo counterpart; however, MUC5B was more homogeneous than that found in vivo. Finally, evidence is presented for an intracellular NH(2)-terminal cleavage of the MUC5B mucins. These studies indicate that the mucins produced by cultured NHTBE cells are similar to those found in human airways, suggesting that this cell culture model is suited for studies of respiratory mucin biosynthesis, processing, and assembly. PMID- 10835317 TI - Increased CD44 expression in human bronchial epithelial repair after damage or plating at low cell densities. AB - We have investigated the effect of mechanical damage, cell density, and cell derived soluble mediators on CD44 expression in a model of bronchial epithelial repair. CD44 (all isoforms) and variant-containing isoforms (CD44v3, CD44v6, and CD44v9) were identified with flow cytometry and immunocytochemistry with image analysis. After mechanical damage, CD44 expression increased up to 500 microm from the wound edge and for up to 48 h in two human bronchial epithelium-derived cell lines, 16HBE14o- and NCI-H292. CD44 expression was unchanged by interferon gamma and increased by <50% by tumor necrosis factor-alpha. To exclude other soluble factors, a Vaseline spacer was used to temporarily divide petri dishes, with cells at high density on one side and those at low density on the other. After the spacer was removed, the cells at low cell density growing in the shared medium expressed up to fourfold higher CD44, although cell proliferation was unchanged. Thus increased CD44 expression at low cell density was not mediated by soluble factors and may reflect functional involvement in cell motility, dedifferentiation, or altered cell-substrate adhesion in epithelial repair. PMID- 10835318 TI - Tyrosine kinase-dependent calcium signaling in airway smooth muscle cells. AB - Contractile agonists may stimulate mitogenic responses in airway smooth muscle by mechanisms that involve tyrosine kinases. The role of contractile agonist-evoked activation of tyrosine kinases in contractile signaling is not clear. We addressed this issue using cultured rat airway smooth muscle cells. In these cells, serotonin (5-HT, 1 microM) caused contraction (quantitated by a decrease in cell area), which was blocked by the tyrosine kinase inhibitor genistein (40 microM). Genistein and tyrphostin 23 (40 and 10 microM, respectively) significantly decreased 5-HT-evoked peak Ca(2+) responses, and the effect of genistein could be observed in the absence of extracellular Ca(2+). The specific inhibitor of mitogen-activated protein kinase kinase PD-98059 (30 microM) had no significant effect on peak Ca(2+) levels. Western analysis of cell extracts revealed that 5-HT caused a significant increase in tyrosine phosphorylation of proteins with molecular masses of approximately 70 kDa within 10 s of stimulation but no measurable tyrosine phosphorylation of the gamma isoform of phospholipase C (PLC-gamma). Tyrosine phosphorylation was inhibited by genistein. Furthermore, genistein (40 microM) significantly attenuated 5-HT-induced inositol phosphate production. We conclude that in airway smooth muscle contractile agonists acting on G protein-coupled receptors may activate tyrosine kinase(s), which in turn modulate calcium signaling by affecting, directly or indirectly, PLC-beta activity. It is unlikely that PLC-gamma or the mitogen-activated protein kinase pathway is involved in Ca(2+) signaling to 5-HT. PMID- 10835319 TI - Autocrine cytokine signaling mediates effects of rhinovirus on airway responsiveness. AB - The airway responses to allergen exposure in allergic asthma are qualitatively similar to those elicited by specific viral respiratory pathogens, most notably rhinovirus (RV), suggesting that the altered airway responsiveness seen in allergic asthma and that elicited by viral respiratory tract infection may share a common underlying mechanism. To the extent that T helper cell type 2 (Th2) cytokines have been implicated in the pathogenesis of allergic asthma, this study examined the potential role(s) of Th2-type cytokines in mediating pro-asthmatic like changes in airway smooth muscle (ASM) responsiveness after inoculation of naive ASM with human RV. Isolated rabbit ASM tissues and cultured human ASM cells were exposed to RV (serotype 16) for 24 h in the absence and presence of monoclonal blocking antibodies (MAbs) or antagonists directed against either the Th2-type cytokines interleukin (IL)-4 and IL-5, intercellular adhesion molecule (ICAM)-1 (the endogenous host receptor for most RVs), or the pleiotropic proinflammatory cytokine IL-1beta. Relative to control (vehicle-treated) tissues, RV-exposed ASM exhibited significantly enhanced isometric contractility to acetylcholine and impaired relaxation to isoproterenol. These pro-asthmatic-like changes in ASM responsiveness were ablated by pretreating the RV-exposed tissues with either IL-5-receptor-alpha blocking antibody or human recombinant IL-1 receptor antagonist, whereas IL-4 neutralizing antibody had no effect. Extended studies further demonstrated that inoculation of ASM cells with RV elicited 1) an increased mRNA expression and release of IL-5 protein, which was inhibited in the presence of anti-ICAM-1 MAb, and 2) an enhanced release of IL-1beta protein, which was inhibited in the presence of IL-5 receptor-alpha antibody. Collectively, these observations provide new evidence demonstrating that RV induced changes in ASM responsiveness are largely attributed to ICAM-1-dependent activation of a cooperative autocrine signaling mechanism involving upregulated IL-5-mediated release of IL-1beta by the RV-exposed ASM itself. PMID- 10835320 TI - Intrinsic ICAM-1/LFA-1 activation mediates altered responsiveness of atopic asthmatic airway smooth muscle. AB - Cell adhesion molecules (CAMs) have been importantly implicated in the pathobiology of the airway responses in allergic asthma, including inflammatory cell recruitment into the lungs and altered bronchial responsiveness. To elucidate the mechanism of CAM-related mediation of altered airway responsiveness in the atopic asthmatic state, the expressions and actions of intercellular adhesion molecule-1 (ICAM-1) and its counterreceptor ligand lymphocyte function associated antigen-1 (LFA-1; i.e., CD11a/CD18) were examined in isolated rabbit airway smooth muscle (ASM) tissues and cultured human ASM cells passively sensitized with sera from atopic asthmatic patients or nonatopic nonasthmatic (control) subjects. Relative to control tissues, the atopic asthmatic sensitized ASM exhibited significantly enhanced maximal contractility to acetylcholine and attenuated relaxation responses to isoproterenol. These proasthmatic changes in agonist responsiveness were ablated by pretreating the atopic sensitized tissues with a monoclonal blocking antibody (MAb) to either ICAM-1 or CD11a, whereas a MAb directed against the related beta(2)-integrin Mac-1 had no effect. Moreover, relative to control tissues, atopic asthmatic sensitized ASM cells displayed an autologously upregulated mRNA and cell surface expression of ICAM-1, whereas constitutive expression of CD11a was unaltered. Extended studies further demonstrated that 1) the enhanced expression and release of soluble ICAM-1 by atopic sensitized ASM cells was prevented when cells were pretreated with an interleukin (IL)-5-receptor-alpha blocking antibody and 2) administration of exogenous IL-5 to naive (nonsensitized) ASM cells induced a pronounced soluble ICAM-1 release from the cells. Collectively, these observations provide new evidence demonstrating that activation of the CAM counterreceptor ligands ICAM-1 and LFA-1, both of which are endogenously expressed in ASM cells, elicits autologously upregulated IL-5 release and associated changes in ICAM-1 expression and agonist responsiveness in atopic asthmatic sensitized ASM. PMID- 10835321 TI - Distinct changes in pulmonary surfactant homeostasis in common beta-chain- and GM CSF-deficient mice. AB - Pulmonary alveolar proteinosis (PAP) is caused by inactivation of either granulocyte-macrophage colony-stimulating factor (GM-CSF) or GM receptor common beta-chain (beta(c)) genes in mice [GM(-/-), beta(c)(-/-)], demonstrating a critical role of GM-CSF signaling in surfactant homeostasis. To distinguish possible phenotypic differences in GM(-/-) and beta(c)(-/-) mice, surfactant metabolism was compared in beta(c)(-/-), GM(-/-), and wild-type mice. Although lung histology in beta(c)(-/-) and GM(-/-) mice was indistinguishable, distinct differences were observed in surfactant phospholipid and surfactant protein concentrations and clearance from lungs of beta(c)(-/-) and GM(-/-) mice. At 1-2 days of age, lung saturated phosphatidylcholine (Sat PC) pool sizes were higher in wild-type, beta(c)(-/-), and GM(-/-) mice compared with wild-type adult mice. In wild-type mice, Sat PC pool sizes decreased to adult levels by 7 days of age; however, Sat PC increased with advancing age in beta(c)(-/-) and GM(-/-) mice. Postnatal changes in Sat PC pool sizes were different in GM(-/-) compared with beta(c)(-/-) mice. After 7 days of age, the increased lung Sat PC pool sizes remained constant in beta(c)(-/-) mice but continued to increase in GM(-/-) mice, so that by 56 days of age, lung Sat PC pools were increased three- and sixfold, respectively, compared with wild-type controls. After intratracheal injection, the percent recovery of [(3)H]dipalmitoylphosphatidylcholine and (125)I recombinant surfactant protein (SP) C was higher in beta(c)(-/-) compared with wild-type mice, reflecting decreased clearance in the receptor-deficient mice. The defect in clearance was significantly more severe in GM(-/-) than in beta(c)( /-) mice. The ratio of SP Sat PC to SP-A, -B, and -C was similar in bronchoalveolar lavage fluid (BALF) from adult mice of all genotypes, but the ratio of SP-D to Sat PC was markedly increased in beta(c)(-/-) and GM(-/-) mice (10- and 5-fold, respectively) compared with wild-type mice. GM-CSF concentrations were increased in BALF but not in serum of beta(c)(-/-) mice, consistent with a pulmonary response to the lack of GM-CSF signaling. The observed differences in surfactant metabolism suggest the presence of alternative clearance mechanisms regulating surfactant homeostasis in beta(c)(-/-) and GM(-/ ) mice and may provide a molecular basis for the range in severity of PAP symptoms. surfactant metabolism; alveolar macrophage; granulocyte-macrophage colony-stimulating factor PMID- 10835322 TI - Augmentation of LTC(4) synthesis in human eosinophils caused by CD3-stimulated Th2-like cells in vitro. AB - We assessed the effect of anti-CD3-stimulated secretion of cultured human Th1- and Th2-like cells on leukotriene C(4) (LTC(4)) secretion in isolated human eosinophils. T helper (Th) cell subsets were generated from human naive CD4(+) T cells cocultured with irradiated human transformed B cells and either recombinant human interleukin (rhIL)-1beta plus rhIL-6 plus rhIL-12 for Th1-like cells or rhIL-1beta plus rhIL-6 plus rhIL-4 for Th2-like cells. Coincubation of eosinophils with 1:5 dilution of Th2-supernatant (Sup) caused an increase in LTC(4) secretion caused by 0.1 microM formyl-Met-Leu-Phe and 5 microg/ml cytochalasin B from 921 +/- 238 to 3,067 +/- 1,462 pg/10(6) eosinophils (P < 0.01). Th1-Sup at the same dilution had no augmenting effect on stimulated secretion of LTC(4) in eosinophils despite substantial concentrations of granulocyte-macrophage colony-stimulating factor (GM-CSF) in the supernatant. Dilution of Th1-Sup caused increased LTC(4) that returned to baseline after immunoabsorption of GM-CSF, suggesting the presence of a possible inhibitory factor. We demonstrate that pretreatment of eosinophils with 1:5 dilution of Th2 Sup but not of Th1-Sup causes substantial augmentation of LTC(4) secretion in vitro and establishes that human Th2 cells cause direct augmentation of LTC(4) secretion within 15-30 min of exposure. PMID- 10835323 TI - Increased lung expansion alters the proportions of type I and type II alveolar epithelial cells in fetal sheep. AB - Type I and type II alveolar epithelial cells (AECs) are derived from the same progenitor cell, but little is known about the factors that regulate their differentiation into separate phenotypes. An alteration in lung expansion alters the proportion type II AECs in the fetal lung, indicating that this may be a regulatory factor. Our aim was to quantify the changes in the proportion of type I and type II AECs caused by increased fetal lung expansion and to provide evidence for transdifferentiation of type II into type I cells. Lung tissue samples were collected from ovine fetuses exposed to increased lung expansion induced by 2, 4, or 10 days of tracheal obstruction (TO). The identities and proportions of AEC types were determined with electron microscopy. The proportion of type II cells was reduced from 28.5 +/- 2.2% in control fetuses to 9.4 +/- 2.3% at 2 days of TO and then to 1.9 +/- 0.8% at 10 days. The proportion of type I AECs was not altered at 2 days of TO (63.1 +/- 2.3%) compared with that of control cells (64.8 +/- 0.5%) but was markedly elevated (to 89.4 +/- 0.9%) at 10 days of TO. The proportion of an intermediate AEC type, which displayed characteristics of both type I and type II cells, increased from 5.7 +/- 1.3% in control fetuses to 23.8 +/- 5.1% by 2 days of TO and was similar to control values at 10 days of TO (7.7 +/- 0.9%). Our data show that increases in fetal lung expansion cause time-dependent changes in the proportion of AEC types, including a transient increase in an intermediate cell type. These data provide the first evidence to support the hypothesis that increases in fetal lung expansion induce differentiation of type II into type I AECs via an intermediate cell type. PMID- 10835324 TI - Modification of transepithelial ion transport in human cultured bronchial epithelial cells by interferon-gamma. AB - Human bronchial epithelial cells were treated in vitro with interferon-gamma or tumor necrosis factor-alpha to assess their effect on transepithelial ion transport. Short-circuit current measurements revealed that Na(+) absorption was markedly inhibited by interferon-gamma (10-1,000 U/ml). The cystic fibrosis transmembrane conductance regulator was also downregulated by interferon-gamma as evident at the protein level and by the decrease in the cAMP-dependent current. On the other hand, interferon-gamma caused an increase of the current elicited by apical UTP application, which is due to the activity of Ca(2+)-dependent Cl(-) channels. Tumor necrosis factor-alpha caused few changes in ion transport. Transepithelial fluid transport was measured in normal and cystic fibrosis cells. At rest, both types of cells showed an amiloride-sensitive fluid absorption that was inhibited by interferon-gamma but not by tumor necrosis factor-alpha. Our results show that interferon-gamma alters the transepithelial ion transport of cultured bronchial cells. This effect may change the ion composition and/or volume of periciliary fluid. PMID- 10835325 TI - Endotoxin priming of the cyclooxygenase-2-thromboxane axis in isolated rat lungs. AB - Enhanced prostanoid generation has been implicated in vascular abnormalities occurring during endotoxemia and sepsis, and the lung is particularly prone to such events. Prostanoids are generated from arachidonic acid (AA) via cyclooxygenase (COX)-1 or -2, both isoenzymes recently demonstrated to be expressed in different lung cell types. Upregulation of COX may underlie the phenomenon that endotoxin [lipopolysaccharide (LPS)]-exposed lungs show markedly enhanced vasoconstrictor responses to secondarily applied stimuli (priming). Isolated rat lungs were perfused with a physiological salt buffer solution in the absence and presence of 1.5% rat plasma and exposed to different concentrations of LPS (1,000 or 10,000 ng/ml) during a 2-h priming period. No change in physiological variables was noted during this period, although enhanced baseline liberation of both thromboxane (Tx) A(2) and PGI(2) as well as of tumor necrosis factor (TNF)-alpha was evident compared with that in control lungs in the absence of LPS. LPS priming caused a significant elevation in AA-induced pulmonary arterial pressure, ventilation pressure, and lung weight gain. Concomitant increased levels of TxA(2) were found in the buffer perfusate. All changes were largely suppressed by three selective, structurally unrelated COX-2 inhibitors (NS-398, DUP-697, and SC-236) in both buffer- and buffer-plasma-perfused lungs. Anti-TNF-alpha neutralizing antibodies were ineffective under conditions of buffer perfusion. In the presence of plasma components, manyfold augmented TNF alpha generation was noted, and anti-TNF-alpha antibodies significantly suppressed the increase in ventilation pressure but not in the vascular pressor response and lung edema formation. We conclude that the propensity of LPS-primed lungs to respond with enhanced vasoconstriction, edema formation, and bronchoconstriction to a secondarily applied stimulus proceeds nearly exclusively via COX-2 and increased Tx formation, with TNF-alpha generation being involved in the change in bronchomotor reactivity in the presence of plasma constituents. In context with recent immunohistological investigations, LPS-induced upregulation of the COX-2-thromboxane synthase axis in vascular and bronchial smooth muscle cells is suggested to underlie these events. PMID- 10835326 TI - Role of calpain in hypoxic inhibition of nitric oxide synthase activity in pulmonary endothelial cells. AB - Pulmonary artery endothelial cells (PAEC) were exposed to normoxia or hypoxia (0% O(2)-95% N(2)-5% CO(2)) in the presence and absence of calpain inhibitor I or calpeptin, after which endothelial nitric oxide synthase (eNOS) activity and protein content were assayed. Exposure to hypoxia decreased eNOS activity but not eNOS protein content. Both calpain inhibitor I and calpeptin prevented the hypoxic decrease of eNOS activity. Incubation of calpain with total membrane preparations of PAEC caused dose-dependent decreases in eNOS activity independent of changes in eNOS protein content. Exposure of PAEC to hypoxia also caused time dependent decreases of heat shock protein 90 (HSP90) that were prevented by calpain inhibitor I and calpeptin. Moreover, the HSP90 content in anti-eNOS antibody-induced immunoprecipitates from hypoxic PAEC lysates was reduced, and repletion of HSP90 reversed the decrease of eNOS activity in these immunoprecipitates. Incubation of PAEC with a specific inhibitor of HSP90 (geldanamycin) mimicked the hypoxic decrease of eNOS activity. These results indicate that the hypoxia-induced reduction in eNOS activity in PAEC is due to a decrease in HSP90 caused by calpain activation. PMID- 10835327 TI - Airway surface liquid composition in mice. AB - Airway surface liquid (ASL) lines the conducting airways of the respiratory tract. We collected small samples of this liquid from the lower tracheae of anesthetized C57BL/6 mice and determined its ionic composition (in mM: 87.2 Na(+), 4.7 K(+), and 57.0 Cl(-)). Intravenous methacholine produced significant increases in the concentrations of Na(+), K(+), and Cl(-) within ASL. A limited analysis of liquid from cystic fibrosis transmembrane conductance regulator (CFTR) knockout mice revealed no significant differences compared with littermate controls; however, Pseudomonas aeruginosa infection led to an increase in the salt concentration of ASL in cftr(+/+) mice. Morphometric measurements of tracheal submucosal gland volume revealed significant differences between inbred mouse strains, corresponding to ease of ASL collection. We conclude that although submucosal glands may be responsible for the production of some ASL, the ionic composition of this liquid is actively regulated by the underlying epithelial cells. PMID- 10835328 TI - Experimental silicosis: a shift to a preferential IFN-gamma-based Th1 response in thoracic lymph nodes. AB - In chronic silicosis, mechanisms leading to lymphocyte activation are still poorly understood, although it is well known that not only the lung but also the draining lymph nodes are affected. In the present study, we investigated T-cell activation by analysis of cytokine expression in the enlarged thoracic lymph nodes of rats 2 mo after an 8-day silica aerosol exposure. In the case of helper T cell (Th) type 1 cytokines, we found a significant increase in interferon (IFN) gamma mRNA expression, whereas interleukin (IL)-2 expression remained unchanged. In contrast, gene transcription for the Th2-type cytokines IL-4 and IL-10 was diminished. In addition, with use of an in vitro lymphocyte-macrophage coculture system, an enhanced IFN-gamma and a reduced IL-10 release were shown with cells from silicotic animals. With regard to IFN-gamma-inducing cytokines, we observed enhanced IL-12 mRNA levels in vivo, whereas IL-18 gene expression was slightly decreased. These data indicate that a persistent shift toward an IFN-gamma dominated type 1 (Th1/cytotoxic T cell type 1) T-cell reaction pattern occurred within the thoracic lymph nodes of silicotic animals. Thus a mutual activation of lymphocytes and macrophages may maintain the chronic inflammatory changes that characterize silicosis. PMID- 10835329 TI - Ontogeny and localization of TGF-beta type I receptor expression during lung development. AB - Transforming growth factor (TGF)-beta is a family of multifunctional cytokines controlling cell growth, differentiation, and extracellular matrix deposition in the lung. The biological effects of TGF-beta are mediated by type I (TbetaR-I) and II (TbetaR-II) receptors. Our previous studies show that the expression of TbetaR-II is highly regulated in a spatial and temporal fashion during lung development. In the present studies, we investigated the temporal-spatial pattern and cellular expression of TbetaR-I during lung development. The expression level of TbetaR-I mRNA in rat lung at different embryonic and postnatal stages was analyzed by Northern blotting. TbetaR-I mRNA was expressed in fetal rat lungs in early development and then decreased as development proceeded. The localization of TbetaR-I in fetal and postnatal rat lung tissues was investigated by using in situ hybridization performed with an antisense RNA probe. TbetaR-I mRNA was present in the mesenchyme and epithelium of gestational day 14 rat lungs. An intense TbetaR-I signal was observed in the epithelial lining of the developing bronchi. In gestational day 16 lungs, the expression of TbetaR-I mRNA was increased in the mesenchymal tissue. The epithelium in both the distal and proximal bronchioles showed a similar level of TbetaR-I expression. In postnatal lungs, TbetaR-I mRNA was detected in parenchymal tissues and blood vessels. We further studied the expression of TbetaR-I in cultured rat lung cells. TbetaR-I was expressed by cultured rat lung fibroblasts, microvascular endothelial cells, and alveolar epithelial cells. These studies demonstrate a differential regulation and localization of TbetaR-I that is different from that of TbetaR-II during lung development. TbetaR-I, TbetaR-II, and TGF-beta isoforms exhibit distinct but overlapping patterns of expression during lung development. This implies a distinct role for TbetaR-I in mediating TGF-beta signal transduction during lung development. PMID- 10835330 TI - Acid aspiration increases sensitivity to increased ambient oxygen concentrations. AB - Previously we have demonstrated that prolonged exposure to 100% ambient oxygen leads to a marked loss in functional lung volume and lung compliance, hypoxemia, and surfactant system abnormalities similar to acute respiratory distress syndrome (ARDS). However, 50% oxygen administration is believed to be safe in most clinical settings. In the present study, we have evaluated the effects of a 24-h exposure to 50% oxygen in rabbits immediately following experimental gastric acid aspiration. Mild hypoxemia, but no changes in mortality, lung volume, lung compliance, surfactant metabolism, or edema formation occurred after 24 h of normoxia postacid aspiration. Conversely, a relatively short (24-h) exposure to 50% oxygen after acid aspiration results in increased pulmonary edema, physical signs of respiratory distress, and mortality, as well as decreased arterial oxygenation, lung volume, lung compliance, and type II alveolar cell surfactant synthesis. These results suggest that acid aspiration alters the "set point" for oxygen toxicity, possibly by "priming" cells through activation of inflammatory pathways. This pathogenic mechanism may contribute to the progression of aspiration pneumonia to ARDS. PMID- 10835331 TI - pH-regulated chloride secretion in fetal lung epithelia. AB - The fetal lung actively transports chloride across the airway epithelium. ClC-2, a pH-activated chloride channel, is highly expressed in the fetal lung and is located on the apical surface of the developing respiratory epithelium. Our goal was to determine whether acidic pH could stimulate chloride secretion in fetal rat distal lung epithelial cells mounted in Ussing chambers. A series of acidic solutions stimulated equivalent short-circuit current (I(eq)) from a baseline of 28 +/- 4.8 (pH 7.4) to 70 +/- 5 (pH 6.2), 114 +/- 12.8 (pH 5.0), and 164 +/- 19.2 (pH 3.8) microA/cm(2). These changes in I(eq) were inhibited by 1 mM cadmium chloride and did not result in large changes in [(3)H]mannitol paracellular flux. Immunofluorescent detection by confocal microscopy revealed that ClC-2 is expressed along the luminal surface of polarized fetal distal lung epithelial cells. These data suggest that the acidic environment of the fetal lung fluid could activate chloride channels contributing to fetal lung fluid production and that the changes in I(eq) seen in these Ussing studies may be due to stimulation of ClC-2. PMID- 10835332 TI - Conditional clara cell ablation reveals a self-renewing progenitor function of pulmonary neuroendocrine cells. AB - The neuroepithelial body (NEB) is a highly dynamic structure that responds to chronic airway injury through hyperplasia of associated pulmonary neuroendocrine (PNE) cells. Although NEB dysplasia is correlated with preneoplastic conditions and PNE cells are thought to serve as a precursor for development of small cell lung carcinoma, mechanisms regulating expansion of the PNE cell population are not well understood. Based on studies performed in animal models, it has been suggested that NEB-associated progenitor cells that are phenotypically distinct from PNE cells contribute to PNE cell hyperplasia. We have previously used a Clara cell-specific toxicant, naphthalene, to induce airway injury in mice and have demonstrated that naphthalene-resistant Clara cells, characterized by their expression of Clara cell secretory protein (CCSP), and PNE cells contribute to airway repair and associated hyperplasia of NEBs. This study was conducted to define the contribution of NEB-associated CCSP-expressing progenitor cells to PNE cell hyperplasia after Clara cell ablation. Transgenic (CCtk) mice were generated in which herpes simplex virus thymidine kinase was expressed within all CCSP expressing cells of the conducting airway epithelium through the use of transcriptional regulatory elements from the mouse CCSP promoter. Chronic administration of ganciclovir (GCV) to CCtk transgenic mice resulted in selective ablation of CCSP-expressing cells within conducting airways. Proliferation and hyperplasia of PNE cells occurred in the absence of detectable proliferation among any other residual airway epithelial cell populations. These results demonstrate that PNE cells function as a self-renewing progenitor population and that NEB-associated Clara cells are not necessary for PNE cell hyperplasia. PMID- 10835333 TI - Molecular responses of rat tracheal epithelial cells to transmembrane pressure. AB - Smooth muscle constriction in asthma causes the airway to buckle into a rosette pattern, folding the epithelium into deep crevasses. The epithelial cells in these folds are pushed up against each other and thereby experience compressive stresses. To study the epithelial cell response to compressive stress, we subjected primary cultures of rat tracheal epithelial cells to constant elevated pressures on their apical surface (i.e., a transmembrane pressure) and examined changes in the expression of genes that are important for extracellular matrix production and maintenance of smooth muscle activation. Northern blot analysis of RNA extracted from cells subjected to transmembrane pressure showed induction of early growth response-1 (Egr-1), endothelin-1, and transforming growth factor beta1 in a pressure-dependent and time-dependent manner. Increases in Egr-1 protein were detected by immunohistochemistry. Our results demonstrate that airway epithelial cells respond rapidly to compressive stresses. Potential transduction mechanisms of transmembrane pressure were also investigated. PMID- 10835334 TI - HO-1 expression in type II pneumocytes after transpulmonary gene delivery. AB - Somatic cell gene transfer is a potentially useful strategy to alter lung function. However, achieving efficient transfer to the alveolar epithelium, especially in smaller animals, has not been demonstrated. In this study, the rat heme oxygenase-1 (HO-1) gene was delivered to the lungs of neonatal mice via transpulmonary injection. A bidirectional promoter construct coexpressing both HO 1 and a luciferase reporter gene was used so that in vivo gene expression patterns could be monitored in real time. HO-1 expression levels were also modulated with doxycycline and assessed in vivo with bioluminescent light transmitted through the tissues from the coregulated luciferase reporter. As a model of oxidative stress and HO-1-mediated protection, groups of animals were exposed to hyperoxia. After gene transfer, elevated levels of HO-1 were detected predominantly in alveolar type II cells by immunocytochemistry. With overexpression of HO-1, increased oxidative injury was observed. Furthermore, this model demonstrated a cell-specific effect of lung HO-1 overexpression in oxidative stress. Specific control of expression for therapeutic genes is possible in vivo. The transpulmonary approach may prove useful in targeting gene expression to cells of the alveolar epithelium or to circumscribed areas of the lung. PMID- 10835335 TI - Reduced lung endothelial angiotensin-converting enzyme activity in Watanabe hyperlipidemic rabbits in vivo. AB - We investigated pulmonary endothelial function in vivo in 12- to 18-mo-old male Watanabe heritable hyperlipidemic (WHHL; n = 7) and age- and sex-matched New Zealand White (n = 8) rabbits. The animals were anesthetized and artificially ventilated, and the chest was opened and put in total heart bypass. The single pass transpulmonary utilizations of the angiotensin-converting enzyme (ACE) substrate [(3)H]benzoyl-Phe-Ala-Pro (BPAP) and the 5'-nucleotidase (NCT) substrate [(14)C]AMP were estimated, and the first-order reaction parameter A(max)/K(m), where A(max) is the product of enzyme mass and the catalytic rate constant and K(m) is the Michaelis-Menten constant, was calculated. BPAP transpulmonary utilization and A(max)/K(m) were reduced in WHHL (1.69 +/- 0.16 vs. 2.9 +/- 0.44 and 599 +/- 69 vs. 987 +/- 153 ml/min in WHHL and control rabbits, respectively; P < 0.05 for both). No differences were observed in the AMP parameters. BPAP K(m) and A(max) values were estimated separately under mixed order reaction conditions. No differences in K(m) values were found (9.79 +/- 1 vs. 9.9 +/- 1.31microM), whereas WHHL rabbit A(max) was significantly decreased (5.29 +/- 0.88 vs. 7. 93 +/- 0.8 micromol/min in WHHL and control rabbits, respectively; P < 0.05). We conclude that the observed pulmonary endothelial ACE activity reduction in WHHL rabbits appears related to a decrease in enzyme mass rather than to alterations in enzyme affinity. PMID- 10835336 TI - Brief hypoxia differentially regulates LPS-induced IL-1beta and TNF-alpha gene transcription in RAW 264.7 cells. AB - Episodes of tissue hypoxia and reoxygenation frequently occur during gram negative bacteremia that progresses to septic shock. However, few studies have evaluated modulation by hypoxia and reoxygenation of the proinflammatory cytokine gene expression that is normally induced by gram-negative bacteremia or endotoxemia. In buffer-perfused organs, hypoxia downregulates Escherichia coli induced expression of tumor necrosis factor (TNF)-alpha and interleukin (IL) 1beta in the liver but upregulates these cytokines in the lungs. To identify molecular mechanisms underlying these events, we investigated the effects of brief (1.5-h) hypoxia on TNF-alpha and IL-1beta expression in cultured RAW 264.7 cells during their continuous exposure to lipopolysaccharide (LPS) endotoxin derived from E. coli (serotype 055:B5) for up to 24 h. IL-1beta and TNF-alpha concentrations in cell lysates and culture supernatants were measured by ELISA, and steady-state mRNA was measured by Northern analysis. LPS-induced IL-1beta synthesis was downregulated by hypoxia at both the protein and mRNA levels despite no change in cellular redox status as measured by levels of GSH. In contrast, LPS-induced TNF-alpha production was unaffected by hypoxia as assessed by cell lysate mRNA and lysate and supernatant protein levels. Nuclear runoff analysis showed that downregulation of IL-1beta gene expression by hypoxia occurred transcriptionally. Allopurinol or catalase treatment did not alter modulation of LPS-induced IL-1beta expression by hypoxia, suggesting that this suppression was not caused by reactive oxygen species. Cycloheximide pretreatment suggested that hypoxia-induced downregulation of IL-1beta expression did not require de novo protein synthesis. PMID- 10835337 TI - Voltage-gated K(+)-channel activity in ovine pulmonary vasculature is developmentally regulated. AB - To examine mechanisms underlying developmental changes in pulmonary vascular tone, we tested the hypotheses that 1) maturation-related changes in the ability of the pulmonary vasculature to respond to hypoxia are intrinsic to the pulmonary artery (PA) smooth muscle cells (SMCs); 2) voltage-gated K(+) (K(v))-channel activity increases with maturation; and 3) O(2)-sensitive Kv2.1 channel expression and message increase with maturation. To confirm that maturational differences are intrinsic to PASMCs, we used fluorescence microscopy to study the effect of acute hypoxia on cytosolic Ca(2+) concentration ([Ca(2+)](i)) in SMCs isolated from adult and fetal PAs. Although PASMCs from both fetal and adult circulations were able to sense an acute decrease in O(2) tension, acute hypoxia induced a more rapid and greater change in [Ca(2+)](i) in magnitude in PASMCs from adult compared with fetal PAs. To determine developmental changes in K(v) channel activity, the effects of the K(+)-channel antagonist 4-aminopyridine (4 AP) were studied on fetal and adult PASMC [Ca(2+)](i). 4-AP (1 mM) caused PASMC [Ca(2+)](i) to increase by 94 +/- 22% in the fetus and 303 +/- 46% in the adult. K(v)-channel expression and mRNA levels in distal pulmonary arteries from fetal, neonatal, and adult sheep were determined through the use of immunoblotting and semiquantitative RT-PCR. Both Kv2.1-channel protein and mRNA expression in distal pulmonary vasculature increased with maturation. We conclude that there are maturation-dependent changes in PASMC O(2) sensing that may render the adult PASMCs more responsive to acute hypoxia. PMID- 10835338 TI - Signaling from Ras to Rac and beyond: not just a matter of GEFs. AB - Members of a family of intracellular molecular switches, the small GTPases, sense modifications of the extracellular environment and transduce them into a variety of homeostatic signals. Among small GTPases, Ras and the Rho family of proteins hierarchically and/or coordinately regulate signaling pathways leading to phenotypes as important as proliferation, differentiation and apoptosis. Ras and Rho-GTPases are organized in a complex network of functional interactions, whose molecular mechanisms are being elucidated. Starting from the simple concept of linear cascades of events (GTPase-->activator--> GTPase), the work of several laboratories is uncovering an increasingly complex scenario in which upstream regulators of GTPases also function as downstream effectors and influence the precise biological outcome. Furthermore, small GTPases assemble into macromolecular machineries that include upstream activators, downstream effectors, regulators and perhaps even final biochemical targets. We are starting to understand how these macromolecular complexes work and how they are regulated and targeted to their proper subcellular localization. Ultimately, the acquisition of a cogent picture of the various levels of integration and regulation in small GTPase-mediated signaling should define the physiology of early signal transduction events and the pathological implication of its subversion. PMID- 10835340 TI - X-ray structures of a novel acid phosphatase from Escherichia blattae and its complex with the transition-state analog molybdate. AB - The structure of Escherichia blattae non-specific acid phosphatase (EB-NSAP) has been determined at 1.9 A resolution with a bound sulfate marking the phosphate binding site. The enzyme is a 150 kDa homohexamer. EB-NSAP shares a conserved sequence motif not only with several lipid phosphatases and the mammalian glucose 6-phosphatases, but also with the vanadium-containing chloroperoxidase (CPO) of Curvularia inaequalis. Comparison of the crystal structures of EB-NSAP and CPO reveals striking similarity in the active site structures. In addition, the topology of the EB-NSAP core shows considerable similarity to the fold of the active site containing part of the monomeric 67 kDa CPO, despite the lack of further sequence identity. These two enzymes are apparently related by divergent evolution. We have also determined the crystal structure of EB-NSAP complexed with the transition-state analog molybdate. Structural comparison of the native enzyme and the enzyme-molybdate complex reveals that the side-chain of His150, a putative catalytic residue, moves toward the molybdate so that it forms a hydrogen bond with the metal oxyanion when the molybdenum forms a covalent bond with NE2 of His189. PMID- 10835339 TI - Receptor recognition by gp130 cytokines. AB - Cytokines of the gp130 family exert their diverse biological effects by formation of stable high affinity transmembrane receptor complexes that are characterized by the presence of the shared transmembrane signalling receptor gp130. Different gp130 ligands form signalling complexes that vary in both composition and stoichiometry. Analysis of the three-dimensional structure of selected ligands and receptor elements indicates that ligands display three topologically conserved receptor recognition epitopes that interact with complementary ligand recognition elements. The composition of the signalling complex and downstream biological responses is defined by the relative affinity of different receptor components for these epitopes. The detailed structure of receptor recognition epitopes indicates that the generation of small molecule cytokine mimetics may be a feasible objective. PMID- 10835341 TI - A novel two-over-two alpha-helical sandwich fold is characteristic of the truncated hemoglobin family. AB - Small hemoproteins displaying amino acid sequences 20-40 residues shorter than (non-)vertebrate hemoglobins (Hbs) have recently been identified in several pathogenic and non-pathogenic unicellular organisms, and named 'truncated hemoglobins' (trHbs). They have been proposed to be involved not only in oxygen transport but also in other biological functions, such as protection against reactive nitrogen species, photosynthesis or to act as terminal oxidases. Crystal structures of trHbs from the ciliated protozoan Paramecium caudatum and the green unicellular alga Chlamydomonas eugametos show that the tertiary structure of both proteins is based on a 'two-over-two' alpha-helical sandwich, reflecting an unprecedented editing of the classical 'three-over-three' alpha-helical globin fold. Based on specific Gly-Gly motifs the tertiary structure accommodates the deletion of the N-terminal A-helix and replacement of the crucial heme-binding F helix with an extended polypeptide loop. Additionally, concerted structural modifications allow burying of the heme group and define the distal site, which hosts a TyrB10, GlnE7 residue pair. A set of structural and amino acid sequence consensus rules for stabilizing the fold and the bound heme in the trHbs homology subfamily is deduced. PMID- 10835342 TI - RAP1 controls rhoptry targeting of RAP2 in the malaria parasite Plasmodium falciparum. AB - Rhoptry associated protein 1 (RAP1) and 2 (RAP2), together with a poorly described third protein RAP3, form the low molecular weight complex within the rhoptries of Plasmodium falciparum. These proteins are thought to play a role in erythrocyte invasion by the extracellular merozoite and are important vaccine candidates. We used gene-targeting technology in P.falciparum blood-stage parasites to disrupt the RAP1 gene, producing parasites that express severely truncated forms of RAP1. Immunoprecipitation experiments suggest that truncated RAP1 species did not complex with RAP2 and RAP3. Consistent with this were the distinct subcellular localizations of RAP1 and 2 in disrupted RAP1 parasites, where RAP2 does not traffic to the rhoptries but is instead located in a compartment that appears related to the lumen of the endoplasmic reticulum. These results suggest that RAP1 is required to localize RAP2 to the rhoptries, supporting the hypothesis that rhoptry biogenesis is dependent in part on the secretory pathway in the parasite. The observation that apparently host protective merozoite antigens are not essential for efficient erythrocyte invasion has important implications for vaccine design. PMID- 10835343 TI - Prohibitins act as a membrane-bound chaperone for the stabilization of mitochondrial proteins. AB - Prohibitins are ubiquitous, abundant and evolutionarily strongly conserved proteins that play a role in important cellular processes. Using blue native electrophoresis we have demonstrated that human prohibitin and Bap37 together form a large complex in the mitochondrial inner membrane. This complex is similar in size to the yeast complex formed by the homologues Phb1p and Phb2p. In yeast, levels of this complex are increased on co-overexpression of both Phb1p and Phb2p, suggesting that these two proteins are the only components of the complex. Pulse-chase experiments with mitochondria isolated from phb1/phb2-null and PHB1/2 overexpressing cells show that the Phb1/2 complex is able to stabilize newly synthesized mitochondrial translation products. This stabilization probably occurs through a direct interaction because association of mitochondrial translation products with the Phb1/2 complex could be demonstrated. The fact that Phb1/2 is a large multimeric complex, which provides protection of native peptides against proteolysis, suggests a functional homology with protein chaperones with respect to their ability to hold and prevent misfolding of newly synthesized proteins. PMID- 10835344 TI - Structural basis for recognition of the translocated intimin receptor (Tir) by intimin from enteropathogenic Escherichia coli. AB - Intimin is a bacterial adhesion molecule involved in intimate attachment of enteropathogenic and enterohaemorrhagic Escherichia coli to mammalian host cells. Intimin targets the translocated intimin receptor (Tir), which is exported by the bacteria and integrated into the host cell plasma membrane. In this study we localized the Tir-binding region of intimin to the C-terminal 190 amino acids (Int190). We have also determined the region's high-resolution solution structure, which comprises an immunoglobulin domain that is intimately coupled to a novel C-type lectin domain. This fragment, which is necessary and sufficient for Tir interaction, defines a new super domain in intimin that exhibits striking structural similarity to the integrin-binding domain of the Yersinia invasin and C-type lectin families. The extracellular portion of intimin comprises an articulated rod of immunoglobulin domains extending from the bacterium surface, conveying a highly accessible 'adhesive tip' to the target cell. The interpretation of NMR-titration and mutagenesis data has enabled us to identify, for the first time, the binding site for Tir, which is located at the extremity of the Int190 moiety. PMID- 10835345 TI - Definitive hematopoietic stem cells first develop within the major arterial regions of the mouse embryo. AB - The aorta-gonad-mesonephros (AGM) region is a potent hematopoietic site within the mammalian embryo body, and the first place from which hematopoietic stem cells (HSCs) emerge. Within the complex embryonic vascular, excretory and reproductive tissues of the AGM region, the precise location of HSC development is unknown. To determine where HSCs develop, we subdissected the AGM into aorta and urogenital ridge segments and transplanted the cells into irradiated adult recipients. We demonstrate that HSCs first appear in the dorsal aorta area. Furthermore, we show that vitelline and umbilical arteries contain high frequencies of HSCs coincident with HSC appearance in the AGM. While later in development and after organ explant culture we find HSCs in the urogenital ridges, our results strongly suggest that the major arteries of the embryo are the most important sites from which definitive HSCs first emerge. PMID- 10835346 TI - Human dolichol-phosphate-mannose synthase consists of three subunits, DPM1, DPM2 and DPM3. AB - Dolichol-phosphate-mannose (DPM) synthase generates mannosyl donors for glycosylphosphatidylinositols, N-glycan and protein O- and C-mannosylation. In Saccharomyces cerevisiae, this enzyme is encoded by DPM1. We reported previously that mammalian DPM synthase contains catalytic DPM1 and regulatory DPM2 subunits, and that DPM1 requires DPM2 for its stable expression in the endoplasmic reticulum. Here we report that human DPM synthase consists of three subunits. The third subunit, DPM3, comprises 92 amino acids associated with DPM1 via its C terminal domain and with DPM2 via its N-terminal portion. The stability of DPM3 was dependent upon DPM2. However, overexpression of DPM3 in Lec15 cells, a null mutant of DPM2, restored the biosynthesis of DPM with an increase in DPM1, indicating that DPM3 directly stabilized DPM1. Therefore, DPM2 stabilizes DPM3 and DPM3 stabilizes DPM1. DPM synthase activity was 10 times higher in the presence of DPM2, indicating that DPM2 also plays a role in the enzymatic reaction. Schizosaccharomyces pombe has proteins that resemble three human subunits; S.pombe DPM3 restored biosynthesis of DPM in Lec15 cells, indicating its orthologous relationship to human DPM3. PMID- 10835347 TI - TREK-1 is a heat-activated background K(+) channel. AB - Peripheral and central thermoreceptors are involved in sensing ambient and body temperature, respectively. Specialized cold and warm receptors are present in dorsal root ganglion sensory fibres as well as in the anterior/preoptic hypothalamus. The two-pore domain mechano-gated K(+) channel TREK-1 is highly expressed within these areas. Moreover, TREK-1 is opened gradually and reversibly by heat. A 10 degrees C rise enhances TREK-1 current amplitude by approximately 7 fold. Prostaglandin E2 and cAMP, which are strong sensitizers of peripheral and central thermoreceptors, reverse the thermal opening of TREK-1 via protein kinase A-mediated phosphorylation of Ser333. Expression of TREK-1 in peripheral sensory neurons as well as in central hypothalamic neurons makes this K(+) channel an ideal candidate as a physiological thermoreceptor. PMID- 10835348 TI - ABC-me: a novel mitochondrial transporter induced by GATA-1 during erythroid differentiation. AB - Transcription factor GATA-1 is essential for normal erythropoiesis. GATA-binding sites are consistently found in promoters or enhancers of genes expressed selectively in erythroid cells. To discover novel GATA-1-regulated genes, we searched for GATA-1-activated transcripts in G1E cells, an erythroid line derived from GATA-1(-) embryonic stem cells. By subtractive analysis, we identified a new ATP-binding cassette (ABC) transporter that is strongly and rapidly induced by GATA-1. This protein, named ABC-me (for ABC-mitochondrial erythroid), localizes to the mitochondrial inner membrane and is expressed at particularly high levels in erythroid tissues of embryos and adults. ABC-me is induced during erythroid maturation in cell lines and primary hematopoietic cells, and its overexpression enhances hemoglobin synthesis in erythroleukemia cells. The ABC proteins participate in diverse physiological processes by coupling ATP hydrolysis to the transport of a variety of substrates across cell membranes. We speculate that ABC me, a newly identified erythroid-expressed ABC superfamily member, may mediate critical mitochondrial transport functions related to heme biosynthesis. PMID- 10835349 TI - The homodimeric ATP-binding cassette transporter LmrA mediates multidrug transport by an alternating two-site (two-cylinder engine) mechanism. AB - The bacterial LmrA protein and the mammalian multidrug resistance P-glycoprotein are closely related ATP-binding cassette (ABC) transporters that confer multidrug resistance on cells by mediating the extrusion of drugs at the expense of ATP hydrolysis. The mechanisms by which transport is mediated, and by which ATP hydrolysis is coupled to drug transport, are not known. Based on equilibrium binding experiments, photoaffinity labeling and drug transport assays, we conclude that homodimeric LmrA mediates drug transport by an alternating two-site transport (two-cylinder engine) mechanism. The transporter possesses two drug binding sites: a transport-competent site on the inner membrane surface and a drug-release site on the outer membrane surface. The interconversion of these two sites, driven by the hydrolysis of ATP, occurs via a catalytic transition state intermediate in which the drug transport site is occluded. The mechanism proposed for LmrA may also be relevant for P-glycoprotein and other ABC transporters. PMID- 10835350 TI - Membrane hyperpolarization and salt sensitivity induced by deletion of PMP3, a highly conserved small protein of yeast plasma membrane. AB - Yeast plasma membranes contain a small 55 amino acid hydrophobic polypeptide, Pmp3p, which has high sequence similarity to a novel family of plant polypeptides that are overexpressed under high salt concentration or low temperature treatment. The PMP3 gene is not essential under normal growth conditions. However, its deletion increases the plasma membrane potential and confers sensitivity to cytotoxic cations, such as Na(+) and hygromycin B. Interestingly, the disruption of PMP3 exacerbates the NaCl sensitivity phenotype of a mutant strain lacking the Pmr2p/Enap Na(+)-ATPases and the Nha1p Na(+)/H(+) antiporter, and suppresses the potassium dependency of a strain lacking the K(+) transporters, Trk1p and Trk2p. All these phenotypes could be reversed by the addition of high Ca(2+) concentration to the medium. These genetic interactions indicate that the major effect of the PMP3 deletion is a hyperpolarization of the plasma membrane potential that probably promotes a non-specific influx of monovalent cations. Expression of plant RCI2A in yeast could substitute for the loss of Pmp3p, indicating a common role for Pmp3p and the plant homologue. PMID- 10835351 TI - Cytohesin-1 regulates beta-2 integrin-mediated adhesion through both ARF-GEF function and interaction with LFA-1. AB - Intracellular signaling pathways, which regulate the interactions of integrins with their ligands, affect a wide variety of biological functions. Here we provide evidence of how cytohesin-1, an integrin-binding protein and guanine nucleotide exchange factor (GEF) for ARF GTPases, regulates cell adhesion. Mutational analyses of the beta-2 cytoplasmic domain revealed that the adhesive function of LFA-1 depends on its interaction with cytohesin-1, unless the integrin is activated by exogenous divalent cations. Secondly, cytohesin-1 induces expression of an extracellular activation epitope of LFA-1, and the exchange factor function is not essential for this activity. In contrast, LFA-1 mediated cell adhesion and spreading on intercellular cell adhesion molecule 1 is strongly inhibited by a cytohesin-1 mutant, which fails to catalyze ARF GDP-GTP exchange in vitro. Thus, cytohesin-1 is involved in the activation of LFA-1, most probably through direct interaction with the integrin, and induces cell spreading by its ARF-GEF activity. We therefore propose that both direct regulation of the integrin and concomitant changes in the membrane topology of adherent T cells are modulated by dissectable functions of cytohesin-1. PMID- 10835352 TI - The conserved phosphoinositide 3-kinase pathway determines heart size in mice. AB - Phosphoinositide 3-kinase (PI3K) has been shown to regulate cell and organ size in Drosophila, but the role of PI3K in vertebrates in vivo is not well understood. To examine the role of PI3K in intact mammalian tissue, we have created and characterized transgenic mice expressing constitutively active or dominant-negative mutants of PI3K in the heart. Cardiac- specific expression of constitutively active PI3K resulted in mice with larger hearts, while dominant negative PI3K resulted in mice with smaller hearts. The increase or decrease in heart size was associated with comparable increase or decrease in myocyte size. Cardiomyopathic changes, such as myocyte necrosis, apoptosis, interstitial fibrosis or contractile dysfunction, were not observed in either of the transgenic mice. Thus, the PI3K pathway is necessary and sufficient to promote organ growth in mammals. PMID- 10835353 TI - Two different but converging messenger pathways to intracellular Ca(2+) release: the roles of nicotinic acid adenine dinucleotide phosphate, cyclic ADP-ribose and inositol trisphosphate. AB - Hormones and neurotransmitters mobilize Ca(2+) from the endoplasmic reticulum via inositol trisphosphate (IP(3)) receptors, but how a single target cell encodes different extracellular signals to generate specific cytosolic Ca(2+) responses is unknown. In pancreatic acinar cells, acetylcholine evokes local Ca(2+) spiking in the apical granular pole, whereas cholecystokinin elicits a mixture of local and global cytosolic Ca(2+) signals. We show that IP(3), cyclic ADP-ribose and nicotinic acid adenine dinucleotide phosphate (NAADP) evoke cytosolic Ca(2+) spiking by activating common oscillator units composed of IP(3) and ryanodine receptors. Acetylcholine activation of these common oscillator units is triggered via IP(3) receptors, whereas cholecystokinin responses are triggered via a different but converging pathway with NAADP and cyclic ADP-ribose receptors. Cholecystokinin potentiates the response to acetylcholine, making it global rather than local, an effect mediated specifically by cyclic ADP-ribose receptors. In the apical pole there is a common early activation site for Ca(2+) release, indicating that the three types of Ca(2+) release channels are clustered together and that the appropriate receptors are selected at the earliest step of signal generation. PMID- 10835354 TI - Notch signalling via RBP-J promotes myeloid differentiation. AB - The expression of Notch receptors on hematopoietic cells and of cognate ligands on bone marrow stromal cells suggests a possible role for Notch signalling in the regulation of hematopoiesis. In order to assess the involvement of Notch1 signalling in myelopoiesis, 32D myeloid progenitor cell lines were engineered to permit the conditional induction of the constitutively active intracellular domain of murine Notch1 (mN1(IC)) by the 4-hydroxytamoxifen-inducible system. The induction of mN1(IC) resulted in accelerated and increased granulocytic differentiation. These effects were observed under growth conditions that support differentiation and, to a lesser degree, under conditions that normally promote self-renewal. Transient transfection of mN1(IC) deletion mutants showed that the differentiation promoting activity correlated with RBP-J transactivation. Furthermore, expression of a transcriptionally active derivative of RBP-J (RBP-J VP16) increased myeloid differentiation. To test further the role of Notch signalling in a physiological context, 32D cells expressing mNotch1 were cultured on fibroblast layers that either expressed or did not express the Notch ligand Jagged1. Similar to the induction of mN1(IC), Jagged1 accelerated granulocytic differentiation of 32D cells. Taken together, our data suggest that activation of mNotch1 promotes myeloid differentiation via RBP-J transactivation. PMID- 10835355 TI - Saccharomyces cerevisiae Ras/cAMP pathway controls post-diauxic shift element dependent transcription through the zinc finger protein Gis1. AB - The Saccharomyces cerevisiae protein kinase Rim15 was identified previously as a component of the Ras/cAMP pathway acting immediately downstream of cAMP-dependent protein kinase (cAPK) to control a broad range of adaptations in response to nutrient limitation. Here, we show that the zinc finger protein Gis1 acts as a dosage-dependent suppressor of the rim15Delta defect in nutrient limitation induced transcriptional derepression of SSA3. Loss of Gis1 results in a defect in transcriptional derepression upon nutrient limitation of various genes that are negatively regulated by the Ras/cAMP pathway (e.g. SSA3, HSP12 and HSP26). Tests of epistasis as well as transcriptional analyses of Gis1-dependent expression indicate that Gis1 acts in this pathway downstream of Rim15 to mediate transcription from the previously identified post-diauxic shift (PDS) element. Accordingly, deletion of GIS1 partially suppresses, and overexpression of GIS1 exacerbates the growth defect of mutant cells that are compromised for cAPK activity. Moreover, PDS element-driven expression, which is negatively regulated by the Ras/cAMP pathway and which is induced upon nutrient limitation, is almost entirely dependent on the presence of Gis1. PMID- 10835356 TI - SCF(beta)(-TrCP) ubiquitin ligase-mediated processing of NF-kappaB p105 requires phosphorylation of its C-terminus by IkappaB kinase. AB - Processing of the p105 precursor to form the active subunit p50 of the NF-kappaB transcription factor is a unique case in which the ubiquitin system is involved in limited processing rather than in complete destruction of the target substrate. A glycine-rich region along with a downstream acidic domain have been demonstrated to be essential for processing. Here we demonstrate that following IkappaB kinase (IkappaK)-mediated phosphorylation, the C-terminal domain of p105 (residues 918-934) serves as a recognition motif for the SCF(beta)(-TrCP) ubiquitin ligase. Expression of IkappaKbeta dramatically increases processing of wild-type p105, but not of p105-Delta918-934. Dominant-negative beta-TrCP inhibits IkappaK-dependent processing. Furthermore, the ligase and wild-type p105 but not p105-Delta918-934 associate physically following phosphorylation. In vitro, SCF(beta)(-TrCP) specifically conjugates and promotes processing of phosphorylated p105. Importantly, the TrCP recognition motif in p105 is different from that described for IkappaBs, beta-catenin and human immunodeficiency virus type 1 Vpu. Since p105-Delta918-934 is also conjugated and processed, it appears that p105 can be recognized under different physiological conditions by two different ligases, targeting two distinct recognition motifs. PMID- 10835357 TI - Crystal structure of the human RXRalpha ligand-binding domain bound to its natural ligand: 9-cis retinoic acid. AB - The pleiotropic effects of active retinoids are transduced by their cognate nuclear receptors, retinoid X receptors (RXRs) and retinoic acid receptors (RARs), which act as transcriptional regulators activated by two stereoisomers of retinoic acid (RA): 9-cis RA (9-cRA) and all-trans RA (a-tRA). Among nuclear receptors, RXR occupies a central position and plays a crucial role in many intracellular signalling pathways as a ubiquitous heterodimerization partner with numerous other members of this superfamily. Whereas RARs bind both isomers, RXRs exclusively bind 9-cRA. The crystal structure of the ligand-binding domain (LBD) of human RXRalpha bound to 9-cRA reveals the molecular basis of this ligand selectivity and allows a comparison of both apo and holo forms of the same nuclear receptor. In the crystal, the receptor is monomeric and exhibits a canonical agonist conformation without direct contacts between the ligand and the transactivation helix H12. Comparison with the unliganded RXRalpha LBD structure reveals the molecular mechanisms of ligand-induced conformational changes and allows us to describe at the atomic level how these changes generate the proper protein interface involved in nuclear receptor-coactivator interaction. PMID- 10835358 TI - The level of DLDB/CHIP controls the activity of the LIM homeodomain protein apterous: evidence for a functional tetramer complex in vivo. AB - The LIM homeodomain (LIM-HD) protein Apterous (Ap) and its cofactor DLDB/CHIP control dorso- ventral (D/V) patterning and growth of Drosophila wing. To investigate the molecular mechanisms of Ap/CHIP function we altered their relative levels of expression and generated mutants in the LIM1, LIM2 and HD domains of Ap, as well as in the LIM-interacting and self-association domains of CHIP. Using in vitro and in vivo assays we found that: (i) the levels of CHIP relative to Ap control D/V patterning; (ii) the LIM1 and LIM2 domains differ in their contributions to Ap function; (iii) Ap HD mutations cause weak dominant negative effects; (iv) overexpression of ChipDeltaSAD mutants mimics Ap lack-of function, and this dominant negative phenotype is caused by titration of Ap because it can be rescued by adding extra Ap; and (v) overexpression of ChipDeltaLID mutants also causes an Ap lack-of-function phenotype, but it cannot be rescued by extra Ap. These results support the model that the Ap-CHIP active complex in vivo is a tetramer. PMID- 10835360 TI - Distribution of acetylated histones resulting from Gal4-VP16 recruitment of SAGA and NuA4 complexes. AB - We analyzed the targeting of histone acetyltransferase (HAT) complexes by DNA binding activators during transcriptional activation and the resulting distribution of acetylated histones. An in vitro competition assay was developed to acetylate and transcribe a nucleosomal array template in the presence of excess non-specific chromatin, which mimics in vivo conditions. Stimulation of transcription from the nucleosomal array template under competitive conditions by the SAGA and NuA4 HAT complexes depended on the presence of the Gal4-VP16 activator, which recognizes sites in the promoter and directly interacts with these HATs. Importantly, the stimulation of transcription by SAGA and NuA4 depended on the presence of Gal4-VP16 during histone acetylation, and Gal4-VP16 bound nucleosomal templates were acetylated preferentially by SAGA and NuA4 relative to the competitor chromatin. While targeting of the SAGA complex led to H3 acetylation of promoter-proximal nucleosomes, targeting of the NuA4 complex led to a broader domain of H4 acetylation of >3 kbp. Thus, either promoter proximal H3 acetylation by SAGA or broadly distributed acetylation of H4 by NuA4 activated transcription from chromatin templates. PMID- 10835359 TI - Solution structure of the MEF2A-DNA complex: structural basis for the modulation of DNA bending and specificity by MADS-box transcription factors. AB - The solution structure of the 33 kDa complex between the dimeric DNA-binding core domain of the transcription factor MEF2A (residues 1-85) and a 20mer DNA oligonucleotide comprising the consensus sequence CTA(A/T)(4)TAG has been solved by NMR. The protein comprises two domains: a MADS-box (residues 1-58) and a MEF2S domain (residues 59-73). Recognition and specificity are achieved by interactions between the MADS-box and both the major and minor grooves of the DNA. A number of critical differences in protein-DNA contacts observed in the MEF2A-DNA complex and the DNA complexes of the related MADS-box transcription factors SRF and MCM1 provide a molecular explanation for modulation of sequence specificity and extent of DNA bending ( approximately 15 versus approximately 70 degrees ). The structure of the MEF2S domain is entirely different from that of the equivalent SAM domain in SRF and MCM1, accounting for the absence of cross-reactivity with other proteins that interact with these transcription factors. PMID- 10835361 TI - Locus specificity determinants in the multifunctional yeast silencing protein Sir2. AB - Yeast SIR2, the founding member of a conserved gene family, acts to modulate chromatin structure in three different contexts: silent (HM) mating-type loci, telomeres and rDNA. At HM loci and telomeres, Sir2p forms a complex with Sir3p and Sir4p. However, Sir2p's role in rDNA silencing is Sir3/4 independent, requiring instead an essential nucleolar protein, Net1p. We describe two novel classes of SIR2 mutations specific to either HM/telomere or rDNA silencing. Despite their opposite effects, both classes of mutations cluster in the same two regions of Sir2p, each of which borders on a conserved core domain. A surprising number of these mutations are dominant. Several rDNA silencing mutants display a Sir2p nucleolar localization defect that correlates with reduced Net1p binding. Although the molecular defect in HM/telomere-specific mutants is unclear, they mimic an age-related phenotype where Sir3p and Sir4p relocalize to the nucleolus. Artificial targeting can circumvent the silencing defect in a subset of mutants from both classes. These results define distinct functional domains of Sir2p and provide evidence for additional Sir2p-interacting factors with locus-specific silencing functions. PMID- 10835362 TI - Involvement of 14-3-3 proteins in nuclear localization of telomerase. AB - Maintenance of telomeres is implicated in chromosome stabilization and cell immortalization. Telomerase, which catalyzes de novo synthesis of telomeres, is activated in germ cells and most cancers. Telomerase activity is regulated by gene expression for its catalytic subunit, TERT, whereas several lines of evidence have suggested a post-translational regulation of telomerase activity. Here we identify the 14-3-3 signaling proteins as human TERT (hTERT)-binding partners. A dominant-negative 14-3-3 redistributed hTERT, which was normally predominant in the nucleus, into the cytoplasm. Consistent with this observation, hTERT-3A, a mutant that could not bind 14-3-3, was localized into the cytoplasm. Leptomycin B, an inhibitor of CRM1/exportin 1-mediated nuclear export, or disruption of a nuclear export signal (NES)-like motif located just upstream of the 14-3-3 binding site in hTERT impaired the cytoplasmic localization of hTERT. Compared with wild-type hTERT, hTERT-3A increased its association with CRM1. 14-3 3 binding was not required for telomerase activity either in vitro or in cell extracts. These observations suggest that 14-3-3 enhances nuclear localization of TERT by inhibiting the CRM1 binding to the TERT NES-like motif. PMID- 10835363 TI - Characterization of the initiation sites of both polarity strands of a viroid RNA reveals a motif conserved in sequence and structure. AB - Viroids replicate through a rolling-circle mechanism in which the infecting circular RNA and its complementary (-) strand are transcribed. The precise site at which transcription starts was investigated for the avocado sunblotch viroid (ASBVd), the type species of the family of viroids with hammerhead ribozymes. Linear ASBVd (+) and (-) RNAs begin with a UAAAA sequence that maps to similar A+U-rich terminal loops in their predicted quasi-rod-like secondary structures. The sequences around the initiation sites of ASBVd, which replicates and accumulates in the chloroplast, are similar to the promoters of a nuclear-encoded chloroplastic RNA polymerase (NEP), supporting the involvement of an NEP-like activity in ASBVd replication. Since RNA folding appears to be kinetically determined, the specific location of both ASBVd initiation sites provides a mechanistic insight into how the nascent ASBVd strands may fold in vivo. The approach used here, in vitro capping and RNase protection assays, may be useful for investigating the initiation sites of other small circular RNA replicons. PMID- 10835364 TI - One protein from two open reading frames: mechanism of a 50 nt translational bypass. AB - Translating ribosomes bypass a 50 nt coding gap in order to fuse the information found in the two open reading frames (ORFs) of bacteriophage T4 gene 60. This study investigates the underlying mechanism by focusing on the competition between initiation of bypassing and termination at the end of the first ORF. While nearly all ribosomes initiate bypassing, no more than 50% resume translation in the second ORF. Two previously described cis-acting stimulatory signals are critical for favoring initiation of bypassing over termination. Genetic analysis of these signals supports a working model in which the first (a stem-loop structure at the junction between the first ORF and the coding gap) interferes with decoding in the A-site, and the second (a stretch of amino acids in the nascent peptide encoded by the first ORF) destabilizes peptidyl-tRNA-mRNA pairing. PMID- 10835365 TI - Expression of the ORF-2 protein of the human respiratory syncytial virus M2 gene is initiated by a ribosomal termination-dependent reinitiation mechanism. AB - Translation of the open reading frame 2 (ORF-2) of the human respiratory syncytial virus M2 gene initiates at one of the three initiation codons located upstream of the termination codon for the first ORF. Replacement of ORF-2 with the major ORF of the chloramphenicol acetyltransferase reporter gene followed by systematic mutagenesis of the putative initiation codons demonstrated the usage of these codons as the translational initiators for ORF-2 expression both in vitro and in vivo. While the efficiency of translation was maintained when only the first and second AUG codons were preserved in vivo, there was no apparent preference in vitro for any of the three codons when only one was present. Mutagenesis studies showed that the location of the termination codon of ORF-1 protein plays a crucial role in directing translation of ORF-2 from the upstream initiation codons in vivo. This indicates that the second ORF is accessed by the ribosomes that are departing from the first ORF and that these ribosomes reinitiate on AUG codons 5' to the point of translation termination. PMID- 10835366 TI - The crystal structures of chloramphenicol phosphotransferase reveal a novel inactivation mechanism. AB - Chloramphenicol (Cm), produced by the soil bacterium Streptomyces venezuelae, is an inhibitor of bacterial ribosomal peptidyltransferase activity. The Cm producing streptomycete modifies the primary (C-3) hydroxyl of the antibiotic by a novel Cm-inactivating enzyme, chloramphenicol 3-O-phosphotransferase (CPT). Here we describe the crystal structures of CPT in the absence and presence of bound substrates. The enzyme is dimeric in a sulfate-free solution and tetramerization is induced by ammonium sulfate, the crystallization precipitant. The tetrameric quaternary structure exhibits crystallographic 222 symmetry and has ATP binding pockets located at a crystallographic 2-fold axis. Steric hindrance allows only one ATP to bind per dimer within the tetramer. In addition to active site binding by Cm, an electron-dense feature resembling the enzyme's product is found at the other subunit interface. The structures of CPT suggest that an aspartate acts as a general base to accept a proton from the 3-hydroxyl of Cm, concurrent with nucleophilic attack of the resulting oxyanion on the gamma phosphate of ATP. Comparison between liganded and substrate-free CPT structures highlights side chain movements of the active site's Arg136 guanidinium group of >9 A upon substrate binding. PMID- 10835367 TI - Origins of minigene-dependent growth inhibition in bacterial cells. AB - The expression of very short open reading frames in Escherichia coli can lead to the inhibition of translation and an arrest in cell growth. Inhibition occurs because peptidyl-tRNA hydrolase fails to recycle sufficiently rapidly peptidyl tRNA released from ribosomes at the stop signal in competition with normal termination, causing starvation for essential species of tRNA. Previous studies have shown that the last sense codon, the strength of the Shine-Dalgarno sequence and the nature and context of the stop codon affect the toxicity associated with mini-gene expression. Here, several important parameters are studied as a function of the length of the mini-gene coding sequence. The rate of peptidyl tRNA drop-off catalysed by translation factors decreases dramatically for peptides longer than a hexamer. The probability that ribosomes recycle without dissociation of the mini-gene mRNA varies strongly with the length of the coding sequence. The peptidyl-tRNA hydrolase rap mutant, unlike the wild-type enzyme, is highly sensitive to the length and sequence of the peptide. Together, these parameters explain the length dependence of mini-gene toxicity. PMID- 10835368 TI - Three-dimensional cryo-electron microscopy localization of EF2 in the Saccharomyces cerevisiae 80S ribosome at 17.5 A resolution. AB - Using a sordarin derivative, an antifungal drug, it was possible to determine the structure of a eukaryotic ribosome small middle dotEF2 complex at 17.5 A resolution by three-dimensional (3D) cryo-electron microscopy. EF2 is directly visible in the 3D map and the overall arrangement of the complex from Saccharomyces cerevisiae corresponds to that previously seen in Escherichia coli. However, pronounced differences were found in two prominent regions. First, in the yeast system the interaction between the elongation factor and the stalk region of the large subunit is much more extensive. Secondly, domain IV of EF2 contains additional mass that appears to interact with the head of the 40S subunit and the region of the main bridge of the 60S subunit. The shape and position of domain IV of EF2 suggest that it might interact directly with P-site bound tRNA. PMID- 10835369 TI - Promiscuous targeting of Bacillus subtilis cell division protein DivIVA to division sites in Escherichia coli and fission yeast. AB - The Bacillus subtilis divIVA gene encodes a coiled-coil protein that shows weak similarity to eukaryotic tropomyosins. The protein is targeted to the sites of cell division and mature cell poles where, in B.subtilis, it controls the site specificity of cell division. Although clear homologues of DivIVA are present only in Gram-positive bacteria, and its role in division site selection is not conserved in the Gram-negative bacterium, Escherichia coli, a DivIVA-green fluorescent protein (GFP) fusion was targeted accurately to division sites and retained at the cell pole in this organism. Remarkably, the same fusion protein was also targeted to nascent division sites and growth zones in the fission yeast Schizosaccharomyces pombe, mimicking the localization of the endogenous tropomyosin-like cell division protein Cdc8p, and F-actin. The results show that a targeting signal for division sites is conserved across the eukaryote prokaryote divide. PMID- 10835370 TI - Selective instability of Orc1 protein accounts for the absence of functional origin recognition complexes during the M-G(1) transition in mammals. AB - To investigate the events leading to initiation of DNA replication in mammalian chromosomes, the time when hamster origin recognition complexes (ORCs) became functional was related to the time when Orc1, Orc2 and Mcm3 proteins became stably bound to hamster chromatin. Functional ORCs, defined as those able to initiate DNA replication, were absent during mitosis and early G(1) phase, and reappeared as cells progressed through G(1) phase. Immunoblotting analysis revealed that hamster Orc1 and Orc2 proteins were present in nuclei at equivalent concentrations throughout the cell cycle, but only Orc2 was stably bound to chromatin. Orc1 and Mcm3 were easily eluted from chromatin during mitosis and early G(1) phase, but became stably bound during mid-G(1) phase, concomitant with the appearance of a functional pre-replication complex at a hamster replication origin. Since hamster Orc proteins are closely related to their human and mouse homologs, the unexpected behavior of hamster Orc1 provides a novel mechanism in mammals for delaying assembly of pre-replication complexes until mitosis is complete and a nuclear structure has formed. PMID- 10835371 TI - Multiple roles of Spo11 in meiotic chromosome behavior. AB - Spo11, a type II topoisomerase, is likely to be required universally for initiation of meiotic recombination. However, a dichotomy exists between budding yeast and the animals Caenorhabditis elegans and Drosophila melanogaster with respect to additional roles of Spo11 in meiosis. In Saccharomyces cerevisiae, Spo11 is required for homolog pairing, as well as axial element (AE) and synaptonemal complex (SC) formation. All of these functions are Spo11 independent in C.elegans and D.melanogaster. We examined Spo11 function in a multicellular fungus, Coprinus cinereus. The C.cinereus spo11-1 mutant shows high levels of homolog pairing and occasionally forms full-length AEs, but no SC. In C.cinereus, Spo11 is also required for maintenance of meiotic chromosome condensation and proper spindle formation. Meiotic progression in spo11-1 is aberrant; late in meiosis basidia undergo programmed cell death (PCD). To our knowledge, this is the first example of meiotic PCD outside the animal kingdom. Ionizing radiation can partially rescue spo11-1 for both AE and SC formation and viable spore production, suggesting that the double-strand break function of Spo11 is conserved and is required for these functions. PMID- 10835372 TI - Partial suppression of the fission yeast rqh1(-) phenotype by expression of a bacterial Holliday junction resolvase. AB - A key stage during homologous recombination is the processing of the Holliday junction, which determines the outcome of the recombination reaction. To dissect the pathways of Holliday junction processing in a eukaryote, we have targeted an Escherichia coli Holliday junction resolvase to the nuclei of fission yeast recombination-deficient mutants and analysed their phenotypes. The resolvase partially complements the UV and hydroxyurea hypersensitivity and associated aberrant mitoses of an rqh1(-) mutant. Rqh1 is a member of the RecQ subfamily of DNA helicases that control recombination particularly during S-phase. Significantly, overexpression of the resolvase in wild-type cells partly mimics the loss of viability, hyper-recombination and 'cut' phenotype of an rqh1(-) mutant. These results indicate that Holliday junctions form in wild-type cells that are normally removed in a non-recombinogenic way, possibly by Rqh1 catalysing their reverse branch migration. We propose that in the absence of Rqh1, replication fork arrest results in the accumulation of Holliday junctions, which can either impede sister chromatid segregation or lead to the formation of recombinants through Holliday junction resolution. PMID- 10835374 TI - The murky origin of Snow White and her T-even dwarfs. PMID- 10835375 TI - Multiple genetic pathways for restarting DNA replication forks in Escherichia coli K-12. AB - In Escherichia coli, the primosome assembly proteins, PriA, PriB, PriC, DnaT, DnaC, DnaB, and DnaG, are thought to help to restart DNA replication forks at recombinational intermediates. Redundant functions between priB and priC and synthetic lethality between priA2::kan and rep3 mutations raise the possibility that there may be multiple pathways for restarting replication forks in vivo. Herein, it is shown that priA2::kan causes synthetic lethality when placed in combination with either Deltarep::kan or priC303:kan. These determinations were made using a nonselective P1 transduction-based viability assay. Two different priA2::kan suppressors (both dnaC alleles) were tested for their ability to rescue the priA-priC and priA-rep double mutant lethality. Only dnaC809,820 (and not dnaC809) could rescue the lethality in each case. Additionally, it was shown that the absence of the 3'-5' helicase activity of both PriA and Rep is not the critical missing function that causes the synthetic lethality in the rep-priA double mutant. One model proposes that replication restart at recombinational intermediates occurs by both PriA-dependent and PriA-independent pathways. The PriA-dependent pathways require at least priA and priB or priC, and the PriA independent pathway requires at least priC and rep. It is further hypothesized that the dnaC809 suppression of priA2::kan requires priC and rep, whereas dnaC809,820 suppression of priA2::kan does not. PMID- 10835376 TI - Methods for estimating gene frequencies and detecting selection in bacterial populations. AB - Recent breakthroughs in molecular technology, most significantly the polymerase chain reaction (PCR) and in situ hybridization, have allowed the detection of genetic variation in bacterial communities without prior cultivation. These methods often produce data in the form of the presence or absence of alleles or genotypes, however, rather than counts of alleles. Using relative allele frequencies from presence-absence data as estimates of population allele frequencies tends to underestimate the frequencies of common alleles and overestimate those of rare ones, potentially biasing the results of a test of neutrality in favor of balancing selection. In this study, a maximum-likelihood estimator (MLE) of bacterial allele frequencies designed for use with presence absence data is derived using an explicit stochastic model of the host infection (or bacterial sampling) process. The performance of the MLE is evaluated using computer simulation and a method is presented for evaluating the fit of estimated allele frequencies to the neutral infinite alleles model (IAM). The methods are applied to estimate allele frequencies at two outer surface protein loci (ospA and ospC) of the Lyme disease spirochete, Borrelia burgdorferi, infecting local populations of deer ticks (Ixodes scapularis) and to test the fit to a neutral IAM. PMID- 10835377 TI - Two classes of sir3 mutants enhance the sir1 mutant mating defect and abolish telomeric silencing in Saccharomyces cerevisiae. AB - Silent information regulators, or Sir proteins, play distinct roles in chromatin mediated transcriptional control at the silent mating-type loci, telomeres, and within the rDNA repeats of Saccharomyces cerevisiae. An unusual collection of sir3 mutant alleles was identified in a genetic screen for enhancers of the sir1 mutant mating-defective phenotype. These sir3-eso mutants, like the sir1 mutant, exhibit little or no mating defects alone, but the sir1 sir3-eso double mutants are essentially nonmating. All of the sir3-eso mutants are defective in telomeric silencing. In some mutants, this phenotype is suppressed by tethering Sir1p to telomeres; other mutants are dominant for mating and telomeric silencing defects. Additionally, several sir3-eso mutants are nonmating in combination with the nat1 N-terminal acetyltransferase mutant. The temperature-sensitive allele sir3-8 has an eso phenotype at permissive temperature, yet acts as a null allele at restrictive temperature due to loss of sir3-8 protein. Sequence analysis showed that eight of the nine sir3-eso alleles have mutations within the N-terminal region that is highly similar to the DNA replication initiation protein Orc1p. Together, these data reveal modular domains for Sir3p and further define its function in silencing chromatin. PMID- 10835378 TI - Identification of a novel allele of SIR3 defective in the maintenance, but not the establishment, of silencing in Saccharomyces cerevisiae. AB - Using a screen for genes that affect telomere function, we isolated sir3-P898R, an allele of SIR3 that reduces telomeric silencing yet does not affect mating. While sir3-P898R mutations cause no detectable mating defect in quantitative assays, they result in synergistic mating defects in combination with mutations such as sir1 that affect the establishment of silencing. In contrast, sir3-P898R in combination with a cac1 mutation, which affects the maintenance of silencing, does not result in synergistic mating defects. MATa sir3-P898R mutants form shmoo clusters in response to alpha-factor, and sir3-P898R strains are capable of establishing silencing at a previously derepressed HML locus with kinetics like that of wild-type SIR3 strains. These results imply that Sir3-P898Rp is defective in the maintenance, but not the establishment of silencing. In addition, overexpression of a C-terminal fragment of Sir3-P898R results in a dominant nonmating phenotype: HM silencing is completely lost at both HML and HMR. Furthermore, HM silencing is most vulnerable to disruption by the Sir3-P898R C terminus immediately after S-phase, the time when new silent chromatin is assembled onto newly replicated DNA. PMID- 10835379 TI - Schizosaccharomyces pombe Ste7p is required for both promotion and withholding of the entry to meiosis. AB - The fission yeast ste7 mutant cannot mate and undergo meiosis, but shows no defect in vegetative growth. We cloned and characterized the ste7 gene. The deduced ste7 gene product (Ste7p) was a protein of 569 amino acids with no significant similarity to other proteins. Transcription of ste7 was induced by nutrient starvation via the function of the transcription factor Ste11p. Disruption of the ste7 gene blocked both conjugation and meiosis, showing that Ste7p plays a positive role in these two processes, probably activating the pheromone signal pathway. Unexpectedly, overexpression of ste7(+) promoted conjugation but inhibited meiosis in wild-type cells. The temperature-sensitive pat1-114 mutant underwent ectopic conjugation at the semirestrictive temperature when its genetic background was ste7(+), whereas the same mutant initiated haploid meiosis when its genetic background was ste7Delta. Two-hybrid analysis suggested that Ste7p interacts physically with both Pat1p and Mei2p, which together constitute the major switch to initiate meiosis. Ste7p tagged with green fluorescent protein accumulated in haploid cells under nutrient starvation until they completed conjugation, but this protein disappeared when they were to enter meiosis. These observations suggest that Ste7p may have a function to suppress the onset of meiosis until the conjugation process has been duly completed. PMID- 10835380 TI - Four chromo-domain proteins of Schizosaccharomyces pombe differentially repress transcription at various chromosomal locations. AB - Transcription is repressed in regions of the fission yeast genome close to centromeres, telomeres, or the silent mating-type cassettes mat2-P and mat3-M. The repression involves the chromo-domain proteins Swi6 and Clr4. We report that two other chromo-domain proteins, Chp1 and Chp2, are also important for these position effects. Chp1 showed a specificity for centromeric regions. Its essentiality for the transcriptional repression of centromeric markers correlates with its importance for chromosome stability. Chp2 appeared more pleiotropic. Its effects on centromeric silencing were less pronounced than those of Chp1, and it participated in telomeric position effects and transcriptional silencing in the mating-type region. We also found that PolII-transcribed genes were repressed when placed in one of the Schizosaccharomyces pombe rDNA clusters, a situation analogous to that in the budding yeast Saccharomyces cerevisiae. Chp2, Swi6, Clr4, and, to a lesser extent, Chp1 participated in that repression. PMID- 10835381 TI - The Saccharomyces cerevisiae mre11(ts) allele confers a separation of DNA repair and telomere maintenance functions. AB - The yeast Mre11 protein participates in important cellular functions such as DNA repair and telomere maintenance. Analysis of structure-function relationships of Mre11 has led to identification of several separation-of-function mutations as well as N- and C-terminal domains essential for Mre11 meiotic and mitotic activities. Previous studies have established that there is a strong correlation between Mre11 DNA repair and telomere maintenance functions and that Mre11-Rad50 Xrs2 complex formation appears to be essential for both of these activities. Here we report that the mre11(ts) allele, previously shown to cause temperature dependent defects in DNA repair and meiosis, confers a temperature-independent telomere shortening, indicating that mre11(ts) is a separation-of-function mutation with respect to DNA repair and telomere maintenance. In a yeast two hybrid system, Mre11(ts) fails to form a homodimer or interact with Rad50 and Xrs2 irrespective of experimental temperatures. These observations collectively suggest that the Pro(162)Ser substitution in Mre11(ts) confers a novel separation of Mre11 mitotic functions. Moreover, we observed that while overexpression of the 5'-3' exonuclease gene EXO1 partially complements the MMS sensitivity of mre11, rad50, and xrs2 null mutants, it has no effect on telomere shortening in these strains. This result provides additional evidence on possible involvement of distinctive mechanisms in DNA repair and telomere maintenance by the Mre11 Rad50-Xrs2 complex. PMID- 10835382 TI - The Saccharomyces cerevisiae centromere protein Slk19p is required for two successive divisions during meiosis. AB - Meiotic cell division includes two separate and distinct types of chromosome segregation. In the first segregational event the sister chromatids remain attached at the centromere; in the second the chromatids are separated. The factors that control the order of chromosome segregation during meiosis have not yet been identified but are thought to be confined to the centromere region. We showed that the centromere protein Slk19p is required for the proper execution of meiosis in Saccharomyces cerevisiae. In its absence diploid cells skip meiosis I and execute meiosis II division. Inhibiting recombination does not correct this phenotype. Surprisingly, the initiation of recombination is apparently required for meiosis II division. Thus Slk19p appears to be part of the mechanism by which the centromere controls the order of meiotic divisions. PMID- 10835383 TI - EXO1 and MSH6 are high-copy suppressors of conditional mutations in the MSH2 mismatch repair gene of Saccharomyces cerevisiae. AB - In Saccharomyces cerevisiae, Msh2p, a central component in mismatch repair, forms a heterodimer with Msh3p to repair small insertion/deletion mismatches and with Msh6p to repair base pair mismatches and single-nucleotide insertion/deletion mismatches. In haploids, a msh2Delta mutation is synthetically lethal with pol3 01, a mutation in the Poldelta proofreading exonuclease. Six conditional alleles of msh2 were identified as those that conferred viability in pol3-01 strains at 26 degrees but not at 35 degrees. DNA sequencing revealed that mutations in several of the msh2(ts) alleles are located in regions with previously unidentified functions. The conditional inviability of two mutants, msh2-L560S pol3-01 and msh2-L910P pol3-01, was suppressed by overexpression of EXO1 and MSH6, respectively. Partial suppression was also observed for the temperature sensitive mutator phenotype exhibited by msh2-L560S and msh2-L910P strains in the lys2-Bgl reversion assay. High-copy plasmids bearing mutations in the conserved EXO1 nuclease domain were unable to suppress msh2-L560S pol3-01 conditional lethality. These results, in combination with a genetic analysis of msh6Delta pol3-01 and msh3Delta pol3-01 strains, suggest that the activity of the Msh2p Msh6p heterodimer is important for viability in the presence of the pol3-01 mutation and that Exo1p plays a catalytic role in Msh2p-mediated mismatch repair. PMID- 10835384 TI - The [KIL-d] element specifically regulates viral gene expression in yeast. AB - The cytoplasmically inherited [KIL-d] element epigenetically regulates killer virus gene expression in Saccharomyces cerevisiae. [KIL-d] results in variegated defects in expression of the M double-stranded RNA viral segment in haploid cells that are "healed" in diploids. We report that the [KIL-d] element is spontaneously lost with a frequency of 10(-4)-10(-5) and reappears with variegated phenotypic expression with a frequency of > or =10(-3). This high rate of loss and higher rate of reappearance is unlike any known nucleic acid replicon but resembles the behavior of yeast prions. However, [KIL-d] is distinct from the known yeast prions in its relative guanidinium hydrochloride incurability and independence of Hsp104 protein for its maintenance. Despite its transmissibility by successive cytoplasmic transfers, multiple cytoplasmic nucleic acids have been proven not to carry the [KIL-d] trait. [KIL-d] epigenetically regulates the expression of the M double-stranded RNA satellite virus genome, but fails to alter the expression of M cDNA. This specificity remained even after a cycle of mating and meiosis. Due to its unique genetic properties and viral RNA specificity, [KIL-d] represents a new type of genetic element that interacts with a viral RNA genome. PMID- 10835385 TI - Loss of Rhb1, a Rheb-related GTPase in fission yeast, causes growth arrest with a terminal phenotype similar to that caused by nitrogen starvation. AB - The Rheb GTPase is most similar in primary sequence to the Ras, Rap, R-Ras, and Ral GTPases, which regulate cell growth and differentiation in many cell types. A likely fission yeast homologue of mammalian Rheb, which we designated Rhb1, was identified by genome sequencing. Our investigation of rhb1 showed that rhb1(-) cells arrested cell growth and division with a terminal phenotype similar to that of nitrogen-starved cells. In particular, cells depleted of Rhb1 arrested as small, round cells with 1N DNA content, arrested more quickly in low-nitrogen medium, and induced expression of fnx1 and mei2 mRNA, two mRNAs that were normally induced by nitrogen starvation. Since mammalian Rheb binds and may regulate Raf-1, a Ras effector, we tested for functional overlap between Ras1 and Rhb1 in fission yeast. This analysis showed that Ras1 overexpression did not suppress rhb1(-) mutant phenotypes, Rhb1 overexpression did not suppress ras1(-) mutant phenotypes, and ras1(-) rhb1(-) double mutants had phenotypes equal to the sum of the corresponding single-mutant phenotypes. Hence, there is no evidence for overlapping functions between Ras1 and Rhb1. On the basis of this study, we hypothesize that Rhb1 negatively regulates entry into stationary phase when extracellular nitrogen levels are adequate for growth. If this hypothesis is correct, then Rhb1 and Ras1 regulate alternative responses to limiting nutrients. PMID- 10835386 TI - Slm9, a novel nuclear protein involved in mitotic control in fission yeast. AB - In the fission yeast Schizosaccharomyces pombe, as in other eukaryotic cells, Cdc2/cyclin B complex is the key regulator of mitosis. Perhaps the most important regulation of Cdc2 is the inhibitory phosphorylation of tyrosine-15 that is catalyzed by Wee1 and Mik1. Cdc25 and Pyp3 phosphatases dephosphorylate tyrosine 15 and activate Cdc2. To isolate novel activators of Cdc2 kinase, we screened synthetic lethal mutants in a cdc25-22 background at the permissive temperature (25 degrees ). One of the genes, slm9, encodes a novel protein of 807 amino acids. Slm9 is most similar to Hir2, the histone gene regulator in budding yeast. Slm9 protein level is constant and Slm9 is localized to the nucleus throughout the cell cycle. The slm9 disruptant is delayed at the G(2)-M transition as indicated by cell elongation and analysis of DNA content. Inactivation of Wee1 fully suppressed the cell elongation phenotype caused by the slm9 mutation. The slm9 mutant is defective in recovery from G(1) arrest after nitrogen starvation. The slm9 mutant is also UV sensitive, showing a defect in recovery from the cell cycle arrest after UV irradiation. PMID- 10835387 TI - ami1, an orthologue of the Aspergillus nidulans apsA gene, is involved in nuclear migration events throughout the life cycle of Podospora anserina. AB - The Podospora anserina ami1-1 mutant was identified as a male-sterile strain. Microconidia (which act as male gametes) form, but are anucleate. Paraphysae from the perithecium beaks are also anucleate when ami1-1 is used as the female partner in a cross. Furthermore, in crosses heterozygous for ami1-1, some crozier cells are uninucleate rather than binucleate. In addition to these nuclear migration defects, which occur at the transition between syncytial and cellular states, ami1-1 causes abnormal distribution of the nuclei in both mycelial filaments and asci. Finally, an ami1-1 strain bearing information for both mating types is unable to self-fertilize. The ami1 gene is an orthologue of the Aspergillus nidulans apsA gene, which controls nuclear positioning in filaments and during conidiogenesis (at the syncytial/cellular transition). The ApsA and AMI1 proteins display 42% identity and share structural features. The apsA gene complements some ami1-1 defects: it increases the percentage of nucleate microconidia and restores self-fertility in an ami1-1 mat+ (mat-) strain. The latter effect is puzzling, since in apsA null mutants sexual reproduction is quite normal. The functional differences between the two genes are discussed with respect to their possible history in these two fungi, which are very distant in terms of evolution. PMID- 10835388 TI - Conidial germination in Aspergillus nidulans requires RAS signaling and protein synthesis. AB - The dormant spores of Aspergillus nidulans become competent for growth and nuclear division in a process called conidial germination. To analyze the molecular details of conidial germination, we developed a genetic screen in which we identified spore germination-deficient mutants that are blocked in this process at the restrictive temperature. These mutants defined eight genes, of which we identified five. Four of the five were directly involved in translation and protein folding, and the fifth showed a high degree of homology to a malonyl CoA synthetase. These results suggest that out of a wide array of processes occurring during conidial germination, translation is essential if germination is to proceed. We also show that conidia containing a mutant-activated form of rasA, the ras homologue in A. nidulans, germinate in the absence of an inducing carbon source, suggesting an important role for rasA signaling in conidial germination. Together these data suggest a model by which a carbon source activates a ras dependent sensory mechanism, inducing translation and leading to conidial germination. This study shows that conidial germination in A. nidulans requires protein synthesis and that the initiation of translation is linked, through an as yet to be determined signaling cascade that includes rasA, to a carbon-source sensing apparatus. PMID- 10835389 TI - Co-expression of the mating-type genes involved in internuclear recognition is lethal in Podospora anserina. AB - In the heterothallic filamentous fungus Podospora anserina, four mating-type genes encoding transcriptional factors have been characterized: FPR1 in the mat+ sequence and FMR1, SMR1, and SMR2 in the alternative mat- sequence. Fertilization is controlled by FPR1 and FMR1. After fertilization, male and female nuclei, which have divided in the same cell, form mat+/mat- pairs during migration into the ascogenous hyphae. Previous data indicate that the formation of mat+/mat- pairs is controlled by FPR1, FMR1, and SMR2. SMR1 was postulated to be necessary for initial development of ascogenous hyphae. In this study, we investigated the transcriptional control of the mat genes by seeking mat transcripts during the vegetative and sexual phase and fusing their promoter to a reporter gene. The data indicate that FMR1 and FPR1 are expressed in both mycelia and perithecia, whereas SMR1 and SMR2 are transcribed in perithecia. Increased or induced vegetative expression of the four mat genes has no effect when the recombined gene is solely in the wild-type strain. However, the combination of resident FPR1 with deregulated SMR2 and overexpressed FMR1 in the same nucleus is lethal. This lethality is suppressed by the expression of SMR1, confirming that SMR1 operates downstream of the other mat genes. PMID- 10835390 TI - Quantitative trait loci affecting components of wing shape in Drosophila melanogaster. AB - Two composite multiple regression-interval mapping analyses were performed to identify candidate quantitative trait loci (QTL) affecting components of wing shape in Drosophila melanogaster defined by eight relative warp-based measures. A recombinant inbred line design was used to map QTL for the shape of two intervein regions in the anterior compartment of the wing, using a high resolution map of retrotransposon insertion sites between Oregon-R and Russian 2b. A total of 35 QTL representing up to 23 different loci were identified, many of which are located near components of the epidermal growth factor-Ras signal transduction pathway that regulates vein vs. intervein decision making and vein placement. Over one-half of the loci were detected in both sexes, and just under one-half were detected at two different growth temperatures. Different loci were found to affect aspects of shape in each intervein region, confirming that the shape of the whole wing should be regarded as a compound trait composed of several developmental units. In addition, a reciprocal backcross design was used to map QTL affecting shape in the posterior compartment of the wings of 831 flies, using a molecular map of 16 allele-specific oligohybridization single nucleotide polymorphism (SNP) markers between two divergent inbred lines. A total of 13 QTL were detected and shown to have generally additive effects on separable components of shape, in both sexes. By contrast, 8 QTL that affected wing size in these backcrosses were nearly dominant in their effects. The results confirm at the genetic level that wing shape is regulated independent of wing size and set up the hypothesis that wing shape is regulated in part through the regulation of the length and positioning of wing veins, involving quantitative regulation of the activity of secreted growth factors. PMID- 10835391 TI - Effect of inversion polymorphism on the neutral nucleotide variability of linked chromosomal regions in Drosophila. AB - Recombination is a main factor determining nucleotide variability in different regions of the genome. Chromosomal inversions, which are ubiquitous in the genus Drosophila, are known to reduce and redistribute recombination, and thus their specific effect on nucleotide variation may be of major importance as an explanatory factor for levels of DNA variation. Here, we use the coalescent approach to study this effect. First, we develop analytical expressions to predict nucleotide variability in old inversion polymorphisms that have reached mutation-drift-flux equilibrium. The effects on nucleotide variability of a new arrangement appearing in the population and reaching a stable polymorphism are then studied by computer simulation. We show that inversions modulate nucleotide variability in a complex way. The establishment of an inversion polymorphism involves a partial selective sweep that eliminates part of the variability in the population. This is followed by a slow convergence to the equilibrium values. During this convergence, regions close to the breakpoints exhibit much lower variability than central regions. However, at equilibrium, regions close to the breakpoints have higher levels of variability and differentiation between arrangements than regions in the middle of the inverted segment. The implications of these findings for overall variability levels during the evolution of Drosophila species are discussed. PMID- 10835392 TI - Heterochromatin protein 1 is required for the normal expression of two heterochromatin genes in Drosophila. AB - The Su(var)2-5 locus, an essential gene in Drosophila, encodes the heterochromatin-associated protein HP1. Here, we show that the Su(var)2-5 lethal period is late third instar. Maternal HP1 is still detectable in first instar larvae, but disappears by third instar, suggesting that developmentally late lethality is probably the result of depletion of maternal protein. We demonstrate that heterochromatic silencing of a normally euchromatic reporter gene is completely lost by third instar in zygotically HP1 mutant larvae, implying a defect in heterochromatin-mediated transcriptional regulation in these larvae. However, expression of the essential heterochromatic genes rolled and light is reduced in Su(var)2-5 mutant larvae, suggesting that reduced expression of essential heterochromatic genes could underlie the recessive lethality of Su(var)2-5 mutations. These results also show that HP1, initially recognized as a transcriptional silencer, is required for the normal transcriptional activation of heterochromatic genes. PMID- 10835393 TI - Molecular analysis of Drosophila eyes absent mutants reveals features of the conserved Eya domain. AB - The eyes absent (eya) gene is critical to eye formation in Drosophila; upon loss of eya function, eye progenitor cells die by programmed cell death. Moreover, ectopic eya expression directs eye formation, and eya functionally synergizes in vivo and physically interacts in vitro with two other genes of eye development, sine oculis and dachshund. The Eya protein sequence, while highly conserved to vertebrates, is novel. To define amino acids critical to the function of the Eya protein, we have sequenced eya alleles. These mutations have revealed that loss of the entire Eya Domain is null for eya activity, but that alleles with truncations within the Eya Domain display partial function. We then extended the molecular genetic analysis to interactions within the Eya Domain. This analysis has revealed regions of special importance to interaction with Sine Oculis or Dachshund. Select eya missense mutations within the Eya Domain diminished the interactions with Sine Oculis or Dachshund. Taken together, these data suggest that the conserved Eya Domain is critical for eya activity and may have functional subregions within it. PMID- 10835394 TI - Analysis of maxillopedia expression pattern and larval cuticular phenotype in wild-type and mutant tribolium. AB - The Tribolium castaneum homeotic gene maxillopedia (mxp) is the ortholog of Drosophila proboscipedia (pb). Here we describe and classify available mxp alleles. Larvae lacking all mxp function die soon after hatching, exhibiting strong transformations of maxillary and labial palps to legs. Hypomorphic mxp alleles produce less severe transformations to leg. RNA interference with maxillopedia double-stranded RNA results in phenocopies of mxp mutant phenotypes ranging from partial to complete transformations. A number of gain-of-function (GOF) mxp alleles have been isolated based on transformations of adult antennae and/or legs toward palps. Finally, we have characterized the mxp expression pattern in wild-type and mutant embryos. In normal embryos, mxp is expressed in the maxillary and labial segments, whereas ectopic expression is observed in some GOF variants. Although mxp and Pb display very similar expression patterns, pb null embryos develop normally. The mxp mutant larval phenotype in Tribolium is consistent with the hypothesis that an ancestral pb-like gene had an embryonic function that was lost in the lineage leading to Drosophila. PMID- 10835396 TI - Histone acetylation and gene expression analysis of sex lethal mutants in Drosophila. AB - The evolution of sex determination mechanisms is often accompanied by reduction in dosage of genes on a whole chromosome. Under these circumstances, negatively acting regulatory genes would tend to double the expression of the genome, which produces compensation of the single-sex chromosome and increases autosomal gene expression. Previous work has suggested that to reduce the autosomal expression to the female level, these dosage effects are modified by a chromatin complex specific to males, which sequesters a histone acetylase to the X. The reduced autosomal histone 4 lysine 16 (H4Lys16) acetylation results in lowered autosomal expression, while the higher acetylation on the X is mitigated by the male specific lethal complex, preventing overexpression. In this report, we examine how mutations in the principal sex determination gene, Sex lethal (Sxl), impact the H4 acetylation and gene expression on both the X and autosomes. When Sxl expression is missing in females, we find that the sequestration occurs concordantly with reductions in autosomal H4Lys16 acetylation and gene expression on the whole. When Sxl is ectopically expressed in Sxl(M) mutant males, the sequestration is disrupted, leading to an increase in autosomal H4Lys16 acetylation and overall gene expression. In both cases we find relatively little effect upon X chromosomal gene expression. PMID- 10835395 TI - A gain-of-function screen for genes that affect the development of the Drosophila adult external sensory organ. AB - The Drosophila adult external sensory organ, comprising a neuron and its support cells, is derived from a single precursor cell via several asymmetric cell divisions. To identify molecules involved in sensory organ development, we conducted a tissue-specific gain-of-function screen. We screened 2293 independent P-element lines established by P. Rorth and identified 105 lines, carrying insertions at 78 distinct loci, that produced misexpression phenotypes with changes in number, fate, or morphology of cells of the adult external sensory organ. On the basis of the gain-of-function phenotypes of both internal and external support cells, we subdivided the candidate lines into three classes. The first class (52 lines, 40 loci) exhibits partial or complete loss of adult external sensory organs. The second class (38 lines, 28 loci) is associated with increased numbers of entire adult external sensory organs or subsets of sensory organ cells. The third class (15 lines, 10 loci) results in potential cell fate transformations. Genetic and molecular characterization of these candidate lines reveals that some loci identified in this screen correspond to genes known to function in the formation of the peripheral nervous system, such as big brain, extra macrochaetae, and numb. Also emerging from the screen are a large group of previously uncharacterized genes and several known genes that have not yet been implicated in the development of the peripheral nervous system. PMID- 10835397 TI - Mitochondrial evidence on the phylogenetic position of caecilians (Amphibia: Gymnophiona). AB - The complete nucleotide sequence (17,005 bp) of the mitochondrial genome of the caecilian Typhlonectes natans (Gymnophiona, Amphibia) was determined. This molecule is characterized by two distinctive genomic features: there are seven large 109-bp tandem repeats in the control region, and the sequence for the putative origin of replication of the L strand can potentially fold into two alternative secondary structures (one including part of the tRNA(Cys)). The new sequence data were used to assess the phylogenetic position of caecilians and to gain insights into the origin of living amphibians (frogs, salamanders, and caecilians). Phylogenetic analyses of two data sets-one combining protein-coding genes and the other combining tRNA genes-strongly supported a caecilian + frog clade and, hence, monophyly of modern amphibians. These two data sets could not further resolve relationships among the coelacanth, lungfishes, and tetrapods, but strongly supported diapsid affinities of turtles. Phylogenetic relationships among a larger set of species of frogs, salamanders, and caecilians were estimated with a mitochondrial rRNA data set. Maximum parsimony analysis of this latter data set also recovered monophyly of living amphibians and favored a frog + salamander (Batrachia) relationship. However, bootstrap support was only moderate at these nodes. This is likely due to an extensive among-site rate heterogeneity in the rRNA data set and the narrow window of time in which the three main groups of living amphibians were originated. PMID- 10835398 TI - Replicative advantage and tissue-specific segregation of RR mitochondrial DNA between C57BL/6 and RR heteroplasmic mice. AB - To investigate the interactions between mtDNA and nuclear genomes, we produced heteroplasmic maternal lineages by transferring the cytoplasts between the embryos of two mouse strains, C57BL/6 (B6) and RR. A total of 43 different nucleotides exist in the displacement-loop (D-loop) region of mtDNA between B6 and RR. Heteroplasmic embryos were reconstructed by electrofusion using a blastomere from a two-cell stage embryo of one strain and an enucleated blastomere from a two-cell stage embryo of the other strain. Equivalent volumes of both types of mtDNAs were detected in blastocyst stage embryos. However, the mtDNA from the RR strain became biased in the progeny, regardless of the source of the nuclear genome. The RR mtDNA population was very high in most of the tissues examined but was relatively low in the brain and the heart. An age related increase of RR mtDNA was also observed in the blood. The RR mtDNAs in the reconstructed embryos and in the embryos collected from heteroplasmic mice showed a different segregation pattern during early embryonic development. These results suggest that the RR mtDNA has a replicative advantage over B6 mtDNA during embryonic development and differentiation, regardless of the type of nuclear genome. PMID- 10835399 TI - Selective genotyping with epistasis can be utilized for a major quantitative trait locus mapping in hypertension in rats. AB - Epistasis used to be considered an obstacle in mapping quantitative trait loci (QTL) despite its significance. Numerous epistases have proved to be involved in quantitative genetics. We established a backcross model that demonstrates a major QTL for hypertension (Ht). Seventy-eight backcrossed rats (BC), derived from spontaneously hypertensive rats (SHR) and normotensive Fischer 344 rats, showed bimodal distribution of systolic blood pressure (BP) values and a phenotypic segregation ratio consistent with 1:1. In this backcross analysis, sarco(endo)plasmic reticulum Ca(2+)-dependent ATPase (Serca) II heterozygotes showed widespread bimodality in frequency distribution of BP values and obviously demonstrated Ht. First, in genome-wide screening, Mapmaker/QTL analysis mapped Ht at a locus between D1Mgh8 and D1Mit4 near Sa in all 78 BC. The peak logarithm of the odds (LOD) score reached 5.3. Second, Serca II heterozygous and homozygous BC were analyzed separately using Mapmaker/QTL. In the 35 Serca II heterozygous BC, the peak LOD score was 3.8 at the same locus whereas it did not reach statistical significance in the 43 Serca II homozygotes. Third, to map Ht efficiently, we selected 18 Serca II heterozygous BC with 9 highest and 9 lowest BP values. In these 18 BC, the peak LOD score reached 8.1. In 17 of the 18, D1Mgh8 genotypes (homo or hetero) qualitatively cosegregated with BP phenotypes (high or low) (P < 0.0001, by chi-square analysis). In conclusion, selective genotyping with epistasis can be utilized for a major QTL mapping near Sa on chromosome 1 in SHR. PMID- 10835400 TI - Narrowing the critical regions for mouse t complex transmission ratio distortion factors by use of deletions. AB - Previously a deletion in mouse chromosome 17, T(22H), was shown to behave like a t allele of the t complex distorter gene Tcd1, and this was attributed to deletion of this locus. Seven further deletions are studied here, with the aim of narrowing the critical region in which Tcd1 must lie. One deletion, T(30H), together with three others, T(31H), T(33H), and T(36H), which extended more proximally, caused male sterility when heterozygous with a complete t haplotype and also enhanced transmission ratio of the partial t haplotype t(6), and this was attributed to deletion of the Tcd1 locus. The deletions T(29H), T(32H), and T(34H) that extended less proximally than T(30H) permitted male fertility when opposite a complete t haplotype. These results enabled narrowing of the critical interval for Tcd1 to between the markers D17Mit164 and D17Leh48. In addition, T(29H) and T(32H) enhanced the transmission ratio of t(6), but significantly less so than T(30H). T(34H) had no effect on transmission ratio. These results could be explained by a new distorter located between the breakpoints of T(29H) and T(34H) (between T and D17Leh66E). It is suggested that the original distorter Tcd1 in fact consists of two loci: Tcd1a, lying between D17Mit164 and D17Leh48, and Tcd1b, lying between T and D17Leh66E. PMID- 10835402 TI - The effects of pollen and seed migration on nuclear-dicytoplasmic systems. I. Nonrandom associations and equilibrium structure with both maternal and paternal cytoplasmic inheritance. AB - We determine the nuclear-dicytoplasmic effects of unidirectional gene flow via pollen and seeds upon a mixed-mating plant population, focusing on nuclear mitochondrial-chloroplast systems where mitochondria are inherited maternally and chloroplasts paternally, as in many conifers. After first delineating the general effects of admixture (via seeds or individuals) on the nonrandom associations in such systems, we derive the full dicytonuclear equilibrium structure, including when disequilibria may be indicators of gene flow. Substantial levels of permanent two- and three-locus disequilibria can be generated in adults by (i) nonzero disequilibria in the migrant pools or (ii) intermigrant admixture effects via different chloroplast frequencies in migrant pollen and seeds. Additionally, three-locus disequilibria can be generated by higher-order intermigrant effects such as different chloroplast frequencies in migrant pollen and seeds coupled with nuclear-mitochondrial disequilibria in migrant seeds, or different nuclear frequencies in migrant pollen and seeds coupled with mitochondrial-chloroplast disequilibria in migrant seeds. Further insight is provided by considering special cases with seed or pollen migration alone, complete random mating or selfing, or migrant pollen and seeds lacking disequilibria or intermigrant admixture effects. The results complete the theoretical foundation for a new method for estimating pollen and seed migration using joint cytonuclear or dicytonuclear data. PMID- 10835401 TI - Physical mapping of male fertility and meiotic drive quantitative trait loci in the mouse t complex using chromosome deficiencies. AB - The t complex spans 20 cM of the proximal region of mouse chromosome 17. A variant form, the t haplotype (t), exists at significant frequencies in wild mouse populations and is characterized by the presence of inversions that suppress recombination with wild-type (+) chromosomes. Transmission ratio distortion and sterility are associated with t and affect males only. It is hypothesized that these phenomena are caused by trans-acting distorter/sterility factors that interact with a responder locus (Tcr(t)) and that the distorter and sterility factors are the same because homozygosity of the distorters causes male sterility. One factor, Tcd1, was previously shown to be amorphic using a chromosome deletion. To overcome limitations imposed by recombination suppression, we used a series of deletions within the t complex in trans to t chromosomes to characterize the Tcd1 region. We find that the distorter activity of Tcd1 is distinct from a linked sterility factor, originally called tcs1. YACs mapped with respect to deletion breakpoints localize tcs1 to a 1.1-Mb interval flanked by D17Aus9 and Tctex1. We present evidence for the existence of multiple proximal t complex regions that exhibit distorter activity. These studies demonstrate the utility of chromosome deletions for complex trait analysis. PMID- 10835403 TI - The effects of pollen and seed migration on nuclear-dicytoplasmic systems. II. A new method for estimating plant gene flow from joint nuclear-cytoplasmic data. AB - A new maximum-likelihood method is developed for estimating unidirectional pollen and seed flow in mixed-mating plant populations from counts of joint nuclear cytoplasmic genotypes. Data may include multiple unlinked nuclear markers with a single maternally or paternally inherited cytoplasmic marker, or with two cytoplasmic markers inherited through opposite parents, as in many conifer species. Migration rate estimates are based on fitting the equilibrium genotype frequencies under continent-island models of plant gene flow to the data. Detailed analysis of their equilibrium structures indicates when each of the three nuclear-cytoplasmic systems allows gene flow estimation and shows that, in general, it is easier to estimate seed than pollen migration. Three-locus nuclear dicytoplasmic data only increase the conditions allowing seed migration estimates; however, the additional dicytonuclear disequilibria allow more accurate estimates of both forms of gene flow. Estimates and their confidence limits for simulated data sets confirm that two-locus data with paternal cytoplasmic inheritance provide better estimates than those with maternal inheritance, while three-locus dicytonuclear data with three modes of inheritance generally provide the most reliable estimates for both types of gene flow. Similar results are obtained for hybrid zones receiving pollen and seed flow from two source populations. An estimation program is available upon request. PMID- 10835404 TI - Variation and selection at the CAULIFLOWER floral homeotic gene accompanying the evolution of domesticated Brassica oleracea. AB - The evolution of plant morphologies during domestication events provides clues to the origin of crop species and the evolutionary genetics of structural diversification. The CAULIFLOWER gene, a floral regulatory locus, has been implicated in the cauliflower phenotype in both Arabidopsis thaliana and Brassica oleracea. Molecular population genetic analysis indicates that alleles carrying a nonsense mutation in exon 5 of the B. oleracea CAULIFLOWER (BoCAL) gene are segregating in both wild and domesticated B. oleracea subspecies. Alleles carrying this nonsense mutation are nearly fixed in B. oleracea ssp. botrytis (domestic cauliflower) and B. oleracea ssp. italica (broccoli), both of which show evolutionary modifications of inflorescence structures. Tests for selection indicate that the pattern of variation at this locus is consistent with positive selection at BoCAL in these two subspecies. This nonsense polymorphism, however, is also present in both B. oleracea ssp. acephala (kale) and B. oleracea ssp. oleracea (wild cabbage). These results indicate that specific alleles of BoCAL were selected by early farmers during the domestication of modified inflorescence structures in B. oleracea. PMID- 10835405 TI - Nucleotide variation at the CHALCONE ISOMERASE locus in Arabidopsis thaliana. AB - An approximately 1.9-kb region encompassing the CHI gene, which encodes chalcone isomerase, was sequenced in 24 worldwide ecotypes of Arabidopsis thaliana (L.) Heynh. and in 1 ecotype of A. lyrata ssp. petraea. There was no evidence for dimorphism at the CHI region. A minimum of three recombination events was inferred in the history of the sampled ecotypes of the highly selfing A. thaliana. The estimated nucleotide diversity theta(TOTAL) = 0.004, theta(SIL) = 0. 005 was on the lower part of the range of the corresponding estimates for other gene regions. The skewness of the frequency spectrum toward an excess of low-frequency polymorphisms, together with the bell-shaped distribution of pairwise nucleotide differences at CHI, suggests that A. thaliana has recently experienced a rapid population growth. Although this pattern could also be explained by a recent selective sweep at the studied region, results from the other studied loci and from an AFLP survey seem to support the expansion hypothesis. Comparison of silent polymorphism and divergence at the CHI region and at the Adh1 and ChiA revealed in some cases a significant deviation of the direct relationship predicted by the neutral theory, which would be compatible with balancing selection acting at the latter regions. PMID- 10835407 TI - Chromosome nondisjunction and instabilities in tapetal cells are affected by B chromosomes in maize. AB - Abnormal mitosis occurs in maize tapetum, producing binucleate cells that later disintegrate, following a pattern of programmed cell death. FISH allowed us to observe chromosome nondisjunction and micronucleus formation in binucleate cells, using DNA probes specific to B chromosomes (B's), knobbed chromosomes, and the chromosome 6 (NOR) of maize. All chromosome types seem to be involved in micronucleus formation, but the B's form more micronuclei than do knobbed chromosomes and knobbed chromosomes form more than do chromosomes without knobs. Micronuclei were more frequent in 1B plants and in a genotype selected for low B transmission rate. Nondisjunction was observed in all types of FISH-labeled chromosomes. In addition, unlabeled bridges and delayed chromatids were observed in the last telophase before binucleate cell formation, suggesting that nondisjunction might occur in all chromosomes of the maize complement. B nondisjunction is known to occur in the second pollen mitosis and in the endosperm, but it was not previously reported in other tissues. This is also a new report of nondisjunction of chromosomes of the normal set (A's) in tapetal cells. Our results support the conclusion that nondisjunction and micronucleus formation are regular events in the process of the tapetal cell death program, but B's strongly increase A chromosome instability. PMID- 10835406 TI - Comparative genetics of disease resistance within the solanaceae. AB - Genomic positions of phenotypically defined disease resistance genes (R genes) and R gene homologues were analyzed in three solanaceous crop genera, Lycopersicon (tomato), Solanum (potato), and Capsicum (pepper). R genes occurred at corresponding positions in two or more genomes more frequently than expected by chance; however, in only two cases, both involving Phytophthora spp., did genes at corresponding positions have specificity for closely related pathogen taxa. In contrast, resistances to Globodera spp., potato virus Y, tobacco mosaic virus, and tomato spotted wilt virus were mapped in two or more genera and did not occur in corresponding positions. Without exception, pepper homologues of the cloned R genes Sw-5, N, Pto, Prf, and I2 were found in syntenous positions in other solanaceous genomes and in some cases also mapped to additional positions near phenotypically defined solanaceous R genes. This detailed analysis and synthesis of all available data for solanaceous R genes suggests a working hypothesis regarding the evolution of R genes. Specifically, while the taxonomic specificity of host R genes may be evolving rapidly, general functions of R alleles (e.g., initiation of resistance response) may be conserved at homologous loci in related plant genera. PMID- 10835408 TI - SHORT INTEGUMENTS 2 promotes growth during Arabidopsis reproductive development. AB - The short integuments 2 (sin2) mutation arrests cell division during integument development of the Arabidopsis ovule and also has subtle pleiotropic effects on both sepal and pistil morphology. Genetic interactions between sin2 and other ovule mutations show that cell division, directionality of growth, and cell expansion represent at least partially independent processes during integument development. Double-mutant analyses also reveal that SIN2 shares functional redundancy with HUELLENLOS in ovule primordium outgrowth and proximal-distal patterning and with TSO1 in promotion of normal morphological development of the four whorls of primary floral organs. All of these observations are consistent with SIN2 being a promoter of growth and cell division during reproductive development, with a primary role in these processes during integument development. On the basis of the floral pleiotropic effects observed in a majority of ovule mutants, including sin2, we postulate a relationship between ovule genes and the evolutionary origin of some processes regulating flower morphology. PMID- 10835409 TI - Genetic drift in an infinite population. The pseudohitchhiking model. AB - Selected substitutions at one locus can induce stochastic dynamics that resemble genetic drift at a closely linked neutral locus. The pseudohitchhiking model is a one-locus model that approximates these effects and can be used to describe the major consequences of linked selection. As the changes in neutral allele frequencies when hitchhiking are rapid, diffusion theory is not appropriate for studying neutral dynamics. A stationary distribution and some results on substitution processes are presented that use the theory of continuous-time Markov processes with discontinuous sample paths. The coalescent of the pseudohitchhiking model is shown to have a random number of branches at each node, which leads to a frequency spectrum that is different from that of the equilibrium neutral model. If genetic draft, the name given to these induced stochastic effects, is a more important stochastic force than genetic drift, then a number of paradoxes that have plagued population genetics disappear. PMID- 10835410 TI - Sojourn times and substitution rate at overdominant and linked neutral loci. AB - The sojourn times until fixation of an overdominant allele were investigated based on the diffusion equation. Furthermore, the rate of accumulation of mutations, or the substitution rate, was predicted from the mean extinction time of a common overdominant allele. The substitution rate calculated theoretically agreed well with that determined by computer simulation. Overdominant selection enhances the polymorphism at linked loci, while its effect on the sojourn times and the substitution rate at linked loci has not been studied yet. To solve these problems, a model that assumed two linked loci, each with infinite alleles, was examined by computer simulation. A decrease in the recombination rate between two loci markedly changed the distribution of sojourn times of a neutral allele. Although overdominant selection obviously increased the sojourn times and the polymorphism at a linked locus, the rate of nucleotide substitution at the neutral locus was not influenced significantly even if complete linkage was assumed. These results suggest that, in regions containing overdominant genes, linked neutral loci will exhibit elevated levels of polymorphism, but their rate of molecular evolution remains that predicted by neutral theory. PMID- 10835411 TI - The effects of Hill-Robertson interference between weakly selected mutations on patterns of molecular evolution and variation. AB - Associations between selected alleles and the genetic backgrounds on which they are found can reduce the efficacy of selection. We consider the extent to which such interference, known as the Hill-Robertson effect, acting between weakly selected alleles, can restrict molecular adaptation and affect patterns of polymorphism and divergence. In particular, we focus on synonymous-site mutations, considering the fate of novel variants in a two-locus model and the equilibrium effects of interference with multiple loci and reversible mutation. We find that weak selection Hill-Robertson (wsHR) interference can considerably reduce adaptation, e.g., codon bias, and, to a lesser extent, levels of polymorphism, particularly in regions of low recombination. Interference causes the frequency distribution of segregating sites to resemble that expected from more weakly selected mutations and also generates specific patterns of linkage disequilibrium. While the selection coefficients involved are small, the fitness consequences of wsHR interference across the genome can be considerable. We suggest that wsHR interference is an important force in the evolution of nonrecombining genomes and may explain the unexpected constancy of codon bias across species of very different census population sizes, as well as several unusual features of codon usage in Drosophila. PMID- 10835412 TI - Inference of population structure using multilocus genotype data. AB - We describe a model-based clustering method for using multilocus genotype data to infer population structure and assign individuals to populations. We assume a model in which there are K populations (where K may be unknown), each of which is characterized by a set of allele frequencies at each locus. Individuals in the sample are assigned (probabilistically) to populations, or jointly to two or more populations if their genotypes indicate that they are admixed. Our model does not assume a particular mutation process, and it can be applied to most of the commonly used genetic markers, provided that they are not closely linked. Applications of our method include demonstrating the presence of population structure, assigning individuals to populations, studying hybrid zones, and identifying migrants and admixed individuals. We show that the method can produce highly accurate assignments using modest numbers of loci-e.g. , seven microsatellite loci in an example using genotype data from an endangered bird species. The software used for this article is available from http://www.stats.ox.ac.uk/ approximately pritch/home. html. PMID- 10835413 TI - The rate of adaptation in asexuals. AB - I study the population genetics of adaptation in asexuals. I show that the rate of adaptive substitution in an asexual species or nonrecombining chromosome region is a bell-shaped function of the mutation rate: at some point, increasing the mutation rate decreases the rate of substitution. Curiously, the mutation rate that maximizes the rate of adaptation depends solely on the strength of selection against deleterious mutations. In particular, adaptation is fastest when the genomic rate of mutation, U, equals the harmonic mean of selection coefficients against deleterious mutations, where we assume that selection for favorable alleles is milder than that against deleterious ones. This simple result is independent of the shape of the distribution of effects among favorable and deleterious mutations, population size, and the action of clonal interference. In the course of this work, I derive an approximation to the probability of fixation of a favorable mutation in an asexual genome or nonrecombining chromosome region in which both favorable and deleterious mutations occur. PMID- 10835414 TI - Gene regulatory networks generating the phenomena of additivity, dominance and epistasis. AB - We show how the phenomena of genetic dominance, overdominance, additivity, and epistasis are generic features of simple diploid gene regulatory networks. These regulatory network models are together sufficiently complex to catch most of the suggested molecular mechanisms responsible for generating dominant mutations. These include reduced gene dosage, expression or protein activity (haploinsufficiency), increased gene dosage, ectopic or temporarily altered mRNA expression, increased or constitutive protein activity, and dominant negative effects. As classical genetics regards the phenomenon of dominance to be generated by intralocus interactions, we have studied two one-locus models, one with a negative autoregulatory feedback loop, and one with a positive autoregulatory feedback loop. To include the phenomena of epistasis and downstream regulatory effects, a model of a three-locus signal transduction network is also analyzed. It is found that genetic dominance as well as overdominance may be an intra- as well as interlocus interaction phenomenon. In the latter case the dominance phenomenon is intimately connected to either feedback-mediated epistasis or downstream-mediated epistasis. It appears that in the intra- as well as the interlocus case there is considerable room for additive gene action, which may explain to some degree the predictive power of quantitative genetic theory, with its emphasis on this type of gene action. Furthermore, the results illuminate and reconcile the prevailing explanations of heterosis, and they support the old conjecture that the phenomenon of dominance may have an evolutionary explanation related to life history strategy. PMID- 10835415 TI - Detecting bottlenecks and selective sweeps from DNA sequence polymorphism. AB - A coalescence-based maximum-likelihood method is presented that aims to (i) detect diversity-reducing events in the recent history of a population and (ii) distinguish between demographic (e.g., bottlenecks) and selective causes (selective sweep) of a recent reduction of genetic variability. The former goal is achieved by taking account of the distortion in the shape of gene genealogies generated by diversity-reducing events: gene trees tend to be more star-like than under the standard coalescent. The latter issue is addressed by comparing patterns between loci: demographic events apply to the whole genome whereas selective events affect distinct regions of the genome to a varying extent. The maximum-likelihood approach allows one to estimate the time and strength of diversity-reducing events and to choose among competing hypotheses. An application to sequence data from an African population of Drosophila melanogaster shows that the bottleneck hypothesis is unlikely and that one or several selective sweeps probably occurred in the recent history of this population. PMID- 10835417 TI - A chromosome-based model for estimating the number of conserved segments between pairs of species from comparative genetic maps PMID- 10835416 TI - A glucose transporter chimera confers a dominant negative glucose starvation phenotype in Saccharomyces cerevisiae. AB - A family of glucose transporters mediates glucose uptake in Saccharomyces cerevisiae. We show that the dominant mutation GSF4-1, which impairs glucose repression of SUC2, results in a nonfunctional chimera of the transporters Hxt1p and Hxt4p. Hxt1/4p inhibits the function of wild-type glucose transporters. Similar mutations may facilitate analysis of the major facilitator superfamily. PMID- 10835418 TI - Mitochondria shape hormonally induced cytoplasmic calcium oscillations and modulate exocytosis. AB - Pituitary gonadotropes transduce hormonal input into cytoplasmic calcium ([Ca(2+)](cyt)) oscillations that drive rhythmic exocytosis of gonadotropins. Using Calcium Green-1 and rhod-2 as optical measures of cytoplasmic and mitochondrial free Ca(2+), we show that mitochondria sequester Ca(2+) and tune the frequency of [Ca(2+)](cyt) oscillations in rat gonadotropes. Mitochondria accumulated Ca(2+) rapidly and in phase with elevations of [Ca(2+)](cyt) after GnRH stimulation or membrane depolarization. Inhibiting mitochondrial Ca(2+) uptake by the protonophore CCCP reduced the frequency of GnRH-induced [Ca(2+)](cyt) oscillations or, occasionally, stopped them. Much of the Ca(2+) that entered mitochondria is bound by intramitochondrial Ca(2+) buffering systems. The mitochondrial Ca(2+) binding ratio may be dynamic because [Ca(2+)](mit) appeared to reach a plateau as mitochondrial Ca(2+) accumulation continued. Entry of Ca(2+) into mitochondria was associated with a small drop in the mitochondrial membrane potential. Ca(2+) was extruded from mitochondria more slowly than it entered, and much of this efflux could be blocked by CGP-37157, a selective inhibitor of mitochondrial Na(+)-Ca(2+) exchange. Plasma membrane capacitance changes in response to depolarizing voltage trains were increased when CCCP was added, showing that mitochondria lower the local [Ca(2+)](cyt) near sites that trigger exocytosis. Thus, we demonstrate a central role for mitochondria in a significant physiological response. PMID- 10835419 TI - Distinct membrane binding properties of N- and C-terminal domains of Escherichia coli SecA ATPase. AB - SecA is a motor protein that drives protein translocation at the Escherichia coli translocon. SecA membrane binding has been shown to occur with high affinity at SecYE and low affinity at anionic phospholipids. To dissect SecA-membrane interaction with reference to SecA structure, the membrane binding properties of N- and C-terminal SecA domains, denoted SecA-N664 and SecA-619C, respectively, were characterized. Remarkably, only SecA-N664 bound to the membrane with high affinity, whereas SecA-619C bound with low affinity in a nonsaturable manner through partitioning with phospholipids. Moreover, SecA-N664 and SecA-619C associated with each other to reconstitute wild type binding affinity. Corroborative results were also obtained from membrane binding competition and subcellular fractionation studies along with binding studies to membranes prepared from strains overproducing SecYE protein. Together, these findings indicate that the specific interaction of SecA with SecYE occurs through its N terminal domain and that the C-terminal domain, although important in SecA membrane cycling at a later stage of translocation, appears to initially assist SecA membrane binding by interaction with phospholipids. These results provide the first evidence for distinct membrane binding characteristics of the two SecA primary domains and their importance for optimal binding activity, and they are significant for understanding SecA dynamics at the translocon. PMID- 10835420 TI - The protein-tyrosine phosphatase SHP-1 binds to and dephosphorylates p120 catenin. AB - A prominent tyrosine-phosphorylated protein of approximately 100 kDa (designated pp100) in epidermal growth factor (EGF)-stimulated A431 cells was found to be a main interaction partner of the protein-tyrosine phosphatase SHP-1 in pull-down experiments with a glutathione S-transferase-SHP-1 fusion protein. Binding was largely mediated by the N-terminal SH2 domain of SHP-1 and apparently direct and independent from the previously described association of SHP-1 with the activated EGF receptor. pp100 was partially purified and identified by mass spectrometric analysis of tryptic fragments, partial amino acid sequencing, and use of authentic antibodies as the 3A isoform of the Armadillo repeat protein superfamily member p120 catenin (p120(ctn)). Different p120(ctn) isoforms expressed in human embryonal kidney 293 cells, exhibited differential binding to SHP-1 that correlated partly with the extent of EGF-dependent p120(ctn) tyrosine phosphorylation. Despite strong phosphorylation, p120(ctn) isoforms 3B and 3AB bound, however, less readily to SHP-1. SHP-1 associated transiently with p120(ctn) in EGF-stimulated A431 cells stably transfected with a tetracycline responsive SHP-1 expression construct, and p120(ctn) exhibited elevated phosphorylation upon a tetracycline-mediated decrease in the SHP-1 level. Functions of p120(ctn), which are regulated by tyrosine phosphorylation, may be modulated by the described SHP-1-p120(ctn) interaction. PMID- 10835421 TI - The basic helix-loop-helix transcription factors myogenin and Id2 mediate specific induction of caveolin-3 gene expression during embryonic development. AB - Caveolin-3 protein is the only member of the caveolin family that shows a unique muscle-specific expression pattern, and loss of its functional activity causes muscular dystrophy. Caveolin-3 mRNA levels are dramatically increased during the formation of myotubes in the C2C12 cell line. In this study, we characterized the human caveolin-3 5'-flanking region. Promoter analyses demonstrate that the proximal E box element serves as a myogenin binding site and is both necessary and sufficient to control caveolin-3 gene transcription. Transient transfection assays indicated that overexpression of myogenin activates caveolin-3 reporter gene expression, whereas Id2 overexpression inhibited caveolin-3 promoter activation by myogenin. A mutant Id2 protein lacking the HLH domain was not capable of suppressing myogenin-mediated activation. Determination of caveolin-3 transcript distribution patterns in vivo revealed that mRNA was first detectable at day 10 of gestation in the developing somites and heart. Caveolin-3 protein in myoblasts and myotubes was expressed in both the plasma membrane and vesicular structures. During skeletal myogenesis the level of Id2, an inhibitor of differentiation, decreases, allowing the induced basic helix-loop-helix transcription factor myogenin to form transcriptionally active heterodimers that bind to the caveolin-3 promoter and thereby mediate its transcription. PMID- 10835422 TI - Regulation of Tiam1 nucleotide exchange activity by pleckstrin domain binding ligands. AB - Rho family GTPases play roles in cytoskeletal organization and cellular transformation. Tiam1 is a member of the Dbl family of guanine nucleotide exchange factors that activate Rho family GTPases. These exchange factors have in common a catalytic Dbl homology and adjacent pleckstrin homology domain. Previous structural studies suggest that the pleckstrin domain, a putative phosphoinositide-binding site, may serve a regulatory function. We identified ascorbyl stearate as a compound that binds to the pleckstrin domain of p120 Ras GTPase-activating protein. Furthermore, ascorbyl stearate appears to be a general pleckstrin domain ligand, perhaps by mimicking an endogenous amphiphilic ligand. Tiam1 nucleotide exchange activity was greatly stimulated by ascorbyl stearate. Certain phosphoinositides also stimulated Tiam1 activity but were less potent than ascorbyl stearate. Tiam1 contains an additional N-terminal pleckstrin domain, but only the C-terminal pleckstrin domain was required for activation. Our results suggest that the pleckstrin domains of Dbl-type proteins may not only be involved in subcellular localization but may also directly regulate the nucleotide exchange activity of an associated Dbl homology domain. In addition, this paper introduces ascorbyl stearate as a pleckstrin domain ligand that can modulate the activity of certain pleckstrin domain-containing proteins. PMID- 10835423 TI - Substrate specificity of alpha(v)beta(3) integrin-mediated cell migration and phosphatidylinositol 3-kinase/AKT pathway activation. AB - The alpha(v)beta(3) integrin has been shown to bind several ligands, including osteopontin and vitronectin. Its role in modulating cell migration and downstream signaling pathways in response to specific extracellular matrix ligands has been investigated in this study. Highly invasive prostate cancer PC3 cells that constitutively express alpha(v)beta(3) adhere and migrate on osteopontin and vitronectin in an alpha(v)beta(3)-dependent manner. However, exogenous expression of alpha(v)beta(3) in noninvasive prostate cancer LNCaP (beta(3)-LNCaP) cells mediates adhesion and migration on vitronectin but not on osteopontin. Activation of alpha(v)beta(3) by epidermal growth factor stimulation is required to mediate adhesion to osteopontin but is not sufficient to support migration on this substrate. We show that alpha(v)beta(3)-mediated cell migration requires activation of the phosphatidylinositol 3-kinase (PI 3-kinase)/protein kinase B (PKB/AKT) pathway since wortmannin, a PI 3-kinase inhibitor, prevents PC3 cell migration on both osteopontin and vitronectin; furthermore, alpha(v)beta(3) engagement by osteopontin and vitronectin activates the PI 3-kinase/AKT pathway. Migration of beta(3)-LNCaP cells on vitronectin also occurs through activation of the PI 3-kinase pathway; however, AKT phosphorylation is not increased upon engagement by osteopontin. Furthermore, phosphorylation of focal adhesion kinase (FAK), known to support cell migration in beta(3)-LNCaP cells, is detected on both substrates. Thus, in PC3 cells, alpha(v)beta(3) mediates cell migration and PI 3-kinase/AKT pathway activation on vitronectin and osteopontin; in beta(3) LNCaP cells, alpha(v)beta(3) mediates cell migration and PI 3-kinase/AKT pathway activation on vitronectin, whereas adhesion to osteopontin does not support alpha(v)beta(3)-mediated cell migration and PI 3-kinase/AKT pathway activation. We conclude therefore that alpha(v)beta(3) exists in multiple functional states that can bind either selectively vitronectin or both vitronectin and osteopontin and that can differentially activate cell migration and intracellular signaling pathways in a ligand-specific manner. PMID- 10835424 TI - Molecular characterization of Vibrio parahaemolyticus vSGLT: a model for sodium coupled sugar cotransporters. AB - The Na(+)/galactose cotransporter (vSGLT) of Vibrio parahaemolyticus, tagged with C-terminal hexahistidine, has been purified to apparent homogeneity by Ni(2+) affinity chromatography and gel filtration. Resequencing the vSGLT gene identified an important correction: the N terminus constitutes an additional 13 functionally essential residues. The mass of His-tagged vSGLT expressed under its native promoter, as determined by electrospray ionization-mass spectrometry (ESI MS), verifies these 13 residues in wild-type vSGLT. A fusion protein of vSGLT and green fluorescent protein, comprising a mass of over 90 kDa, was also successfully analyzed by ESI-MS. Reconstitution of purified vSGLT yields proteoliposomes active in Na(+)-dependent galactose uptake, with sugar preferences (galactose > glucose > fucose) reflecting those of wild-type vSGLT in vivo. Substrates are transported with apparent 1:1 stoichiometry and apparent K(m) values of 129 mm (Na(+)) and 158 microm (galactose). Freeze-fracture electron microscopy of functional proteoliposomes shows intramembrane particles of a size consistent with vSGLT existing as a monomer. We conclude that vSGLT is a suitable model for the study of sugar cotransporter mechanisms and structure, with potential applicability to the larger SGLT family of important sodium:solute cotransporters. It is further demonstrated that ESI-MS is a powerful tool for the study of proteomics of membrane transporters. PMID- 10835425 TI - The interferon- and differentiation-inducible p202a protein inhibits the transcriptional activity of c-Myc by blocking its association with Max. AB - p202a is a murine protein that is induced during the fusion of myoblasts to myotubes and can also be induced by interferon. Even 2-3-fold overexpression of p202a in cells retards proliferation. p202a was shown to modulate transcription by binding, and inhibiting the activity of several transcription factors including c-Fos, c-Jun, AP-2, E2F1, E2F4, NF-kappaB, MyoD, and myogenin. Here we report that p202a also bound the c-Myc protein in vitro and in vivo; the C terminal p202a b segment bound the C-terminal basic region helix-loop-helix leucine zipper (bHLHLZ) region of c-Myc. The transfection of a p202a expression plasmid inhibited the c-Myc-dependent expression of reporter plasmids in transient assays; moreover, overexpression of p202a in stable cell lines decreased the endogenous levels of mRNAs whose expression is driven by c-Myc. These effects of p202a are consistent with our finding that the binding of p202a to c-Myc inhibited the binding of c-Myc to Max in vitro and in vivo. p202a also inhibited the c-Myc-induced anchorage-independent growth and apoptosis of Rat-1 cells. The inhibition of c-Myc-dependent transcription, proliferation, and apoptosis by p202a is in line with the involvement of p202a in differentiation. PMID- 10835426 TI - CalDAG-GEFIII activation of Ras, R-ras, and Rap1. AB - We characterized a novel guanine nucleotide exchange factor (GEF) for Ras family G proteins that is highly homologous to CalDAG-GEFI, a GEF for Rap1 and R-Ras, and to RasGRP/CalDAG-GEFII, a GEF for Ras and R-Ras. This novel GEF, referred to as CalDAG-GEFIII, increased the GTP/GDP ratio of Ha-Ras, R-Ras, and Rap1 in 293T cells. CalDAG-GEFIII promoted the guanine nucleotide exchange of Ha-Ras, R-Ras, and Rap1 in vitro also, indicating that CalDAG-GEFIII exhibited the widest substrate specificity among the known GEFs for Ras family G proteins. Expression of CalDAG-GEFIII was detected in the glial cells of the brain and the glomerular mesangial cells of the kidney by in situ hybridization. CalDAG-GEFIII activated ERK/MAPK most efficiently, followed by CalDAG-GEFII and CalDAG-GEFI in 293T cells. JNK activation was most prominent in cells expressing CalDAG-GEFII, followed by CalDAG-GEFIII and CalDAG-GEFI. Expression of CalDAG-GEFIII induced neuronal differentiation of PC12 cells and anchorage-independent growth of Rat1A cells less efficiently than did CalDAG-GEFII. Thus, co-activation of Rap1 by CalDAG-GEFIII apparently attenuated Ras-MAPK-dependent neuronal differentiation and cellular transformation. Altogether, CalDAG-GEFIII activated a broad range of Ras family G proteins and exhibited a biological activity different from that of either CalDAG-GEFI or CalDAG-GEFII. PMID- 10835427 TI - Hepatitis B virus X protein inhibits transforming growth factor-beta -induced apoptosis through the activation of phosphatidylinositol 3-kinase pathway. AB - Transforming growth factor-beta (TGF-beta) is a potent inducer of apoptosis in Hep 3B cells. This work investigated how hepatitis B virus X protein (HBx) affects TGF-beta-induced apoptosis. Trypan blue exclusion and colony formation assays revealed that HBx increased the ID(50) toward TGF-beta. In the presence of HBx, TGF-beta-induced DNA laddering was decreased, indicating that HBx had the ability to block TGF-beta-induced apoptosis. Furthermore, HBx did not alter the expression levels of type I and type II TGF-beta receptors. HBx did not affect TGF-beta-induced activation of promoter activities of the plasminogen activator inhibitor-1 (PAI-1) gene. These results indicate that HBx interferes with only a subset of TGF-beta activity. In the presence of phosphatidylinositol (PI) 3 kinase inhibitors, wortmannin or LY294002, the HBx-mediated inhibitory effect on TGF-beta-induced apoptosis was alleviated. In addition, the tyrosine phosphorylation levels of the regulatory subunit p85 of phosphatidylinositol 3 kinase (PI 3-kinase) and PI 3-kinase activity were elevated in stable clones with HBx expression. Transactivation-deficient mutants of HBx lost their ability to inhibit TGF-beta-induced apoptosis. Phosphorylation of the p85 subunit of PI 3 kinase and Akt, a downstream target of PI 3-kinase, was not observed in stable clones with transactivation-deficient HBx mutant's expression. Thus, the anti apoptotic effect of HBx against TGF-beta can be mediated through the activation of the PI 3-kinase signaling pathway, and the transactivation function of HBx is required for its anti-apoptosis activity. PMID- 10835428 TI - Calbindin-D28k is expressed in osteoblastic cells and suppresses their apoptosis by inhibiting caspase-3 activity. AB - The rate of osteoblast apoptosis is a critical determinant of the rate of bone formation. Because the calcium-binding protein calbindin-D(28k) has anti apoptotic properties in neuronal cells and lymphocytes, we searched for the presence of this protein in osteoblastic cells and investigated whether it can modify their response to proapoptotic signals. Calbindin-D(28K) was expressed at low levels in several osteoblastic cell lines and at high levels in primary cultures of murine osteoblastic cells. Transient transfection of rat calbindin D(28k) cDNA blocked tumor necrosis factor alpha (TNFalpha)-induced apoptosis in osteoblastic MC3T3-E1 cells, as determined by cell viability and nuclear morphology of cells cotransfected with the green fluorescent protein targeted to the nucleus, whereas transfection of the empty vector had no effect. Calbindin D(28k) levels in several stably transfected MC3T3-E1 lines were directly related to protection from TNFalpha-induced apoptosis. Purified rat calbindin-D(28k) markedly reduced the activity of caspase-3, a critical molecule for the degradation phase of apoptosis, in a cell-free assay. In addition, cell extracts from MC3T3-E1 cells expressing high levels of calbindin-D(28k) decreased caspase 3 activity, compared with extracts from vector-transfected cells. This effect was apparently unrelated to the calcium binding properties of calbindin, as chelation of calcium by EGTA or addition of other calcium-binding proteins such as calbindin-D(9k), S100, calmodulin, and osteocalcin, did not affect caspase-3 activity. Last, calbindin-D(28k) interacts with the active form of caspase-3 as demonstrated by a GST pull-down assay. These results demonstrate that calbindin D(28k) is a biosynthetic product of osteoblasts with a role in the regulation of apoptosis. They also reveal that the antiapoptotic properties of calbindin-D(28k) may result not only from calcium buffering but also from the ability of the protein to interact with and to inhibit caspase-3 activity, a property that is independent of its calcium binding capability. PMID- 10835429 TI - Smoothened activates Galphai-mediated signaling in frog melanophores. AB - The 7-pass transmembrane protein Smoothened was investigated for its ability to act as a G-protein-coupled receptor in Xenopus laevis melanophores. A plasmid containing the human Smoothened cDNA insert was transfected into immortalized frog pigment cells. Cells expressing the protein showed a phenotype of persistent pigment aggregation, a hallmark of constitutive Galpha(i) activation. Smoothened mediated pigment aggregation was reversed by treatment with pertussis toxin or by co-expression with dominant negative Galpha(i). The ability of melanophores to express functional Smoothened was also determined by its co-expression with the twelve-pass transmembrane protein, Patched. Patched blocked Smoothened-mediated melanosome aggregation in a dose-dependent manner, consistent with its physiological role as an inhibitor of Smoothened. That the reconstituted Patched Smoothened receptor complex functions normally in pigment cells was demonstrated by co-transfection with the activating ligand, Sonic hedgehog, as well as by direct application of the recombinant Sonic hedgehog protein. Sonic hedgehog reversed Patched-mediated inhibition of Smoothened and induced pigment aggregation. The findings demonstrate that the human Sonic hedgehog receptor complex can be functionally reconstituted in melanophores and that it is capable of transmembrane signaling by utilizing endogenous Galpha(i). PMID- 10835430 TI - Ligand-independent dimerization activates the stress response kinases IRE1 and PERK in the lumen of the endoplasmic reticulum. AB - IRE1 and PERK are type I transmembrane serine/threonine protein kinases that are activated by unfolded proteins in the endoplasmic reticulum (ER) to signal adaptive responses. IRE1 is present in all eukaryotic cells and signals the unfolded protein response through its kinase and endoribonuclease activities. PERK signals phosphorylation of a translation initiation factor to inhibit protein synthesis in higher eukaryotic cells but is absent in the Saccharomyces cerevisiae genome. The amino acid sequences of the amino-terminal ER luminal domains (NLDs) from IRE1 and PERK display limited homology and have diverged among species. In this study, we have demonstrated that the NLD of yeast Ire1p is required for signaling. However, the NLDs from human IRE1alpha and murine IRE1beta and the Caenorhabditis elegans IRE1 and PERK function as replacements for the S. cerevisiae Ire1p-NLD to signal the unfolded protein response. Replacement of the Ire1p-NLD with a functional leucine zipper dimerization motif yielded a constitutively active kinase that surprisingly was further activated by ER stress. These results demonstrate that ER stress-induced dimerization of the NLD is sufficient for IRE1 and PERK activation and is conserved through evolution. We propose that ligand-independent activation of IRE1 and PERK permits homodimerization upon accumulation of unfolded proteins in the lumen of the ER. PMID- 10835431 TI - Functional importance of the Ala(116)-Pro(136) region in the calcium-sensing receptor. Constitutive activity and inverse agonism in a family C G-protein coupled receptor. AB - The calcium-sensing receptor (CaR) belongs to family C of the G-protein-coupled receptor superfamily. To date 14 activating mutations in CaR showing increased sensitivity to Ca(2+) have been identified in humans with autosomal dominant hypocalcemia. Four of these activating mutations are found in the Ala(116) Pro(136) region of CaR, indicating that this part of the receptor is particularly sensitive to mutation-induced activation. This region was subjected to random saturation mutagenesis, and 219 mutant receptor clones were isolated and screened pharmacologically in a high throughput screening assay. Selected mutants were characterized further in an inositol phosphate assay. The vast majority of the mutants tested displayed an increased affinity for Ca(2+). Furthermore, 21 of the mutants showed increased basal activity in the absence of agonist. This constitutive activity was not diminished when the mutations were transferred to a chimeric receptor Ca/1a consisting of the amino-terminal domain of the CaR and the 7 transmembrane and intracellular domains of the metabotropic glutamate receptor mGluR1a. CPCCOEt, a noncompetitive antagonist acting at the 7 transmembrane domain of mGluR1a, suppressed the elevated basal response of the constitutively activated Ca/1a mutants demonstrating inverse agonist activity of CPCCOEt. Taken together, our results demonstrate that the Ala(116)-Pro(136) region is of key importance for the maintenance of the inactive conformation of CaR. PMID- 10835433 TI - Nine years later: farewell and welcome PMID- 10835432 TI - Ethanol regulates calcium channel subunits by protein kinase C delta -dependent and -independent mechanisms. AB - Chronic exposure to ethanol increases the number of functional L-type voltage gated calcium channels in neural cells. In PC12 cells, this adaptive response is mediated by protein kinase C delta (PKCdelta), but the mechanisms by which this occurs are not known. Since expression of several different calcium channel subunits can increase the abundance of functional L-type channels, we sought to identify which subunits are regulated by ethanol. Incubation of PC12 cells with 120-150 mm ethanol for 6 days increased levels of alpha(1C), alpha(2), and beta(1b) subunit immunoreactivity in cell membranes and selectively increased the abundance of mRNA encoding the alpha(1C-1) splice variant of alpha(1C). In cells expressing a fragment of PKCdelta (deltaV1) that selectively inhibits PKCdelta, there was no increase in membrane-associated alpha(1C), alpha(2), and beta(1b) immunoreactivity following chronic ethanol exposure. However, ethanol still increased levels of alpha(1C-1) mRNA in these cells. These results indicate that ethanol increases the abundance of L-type channels by at least two mechanisms; one involves increases in mRNA encoding a splice variant of alpha(1C) and the other is post-transcriptional, rate-limiting, and requires PKCdelta. PMID- 10835434 TI - Treatment-induced cortical reorganization after stroke in humans. AB - BACKGROUND AND PURPOSE: Injury-induced cortical reorganization is a widely recognized phenomenon. In contrast, there is almost no information on treatment induced plastic changes in the human brain. The aim of the present study was to evaluate reorganization in the motor cortex of stroke patients that was induced with an efficacious rehabilitation treatment. METHODS: We used focal transcranial magnetic stimulation to map the cortical motor output area of a hand muscle on both sides in 13 stroke patients in the chronic stage of their illness before and after a 12-day-period of constraint-induced movement therapy. RESULTS: Before treatment, the cortical representation area of the affected hand muscle was significantly smaller than the contralateral side. After treatment, the muscle output area size in the affected hemisphere was significantly enlarged, corresponding to a greatly improved motor performance of the paretic limb. Shifts of the center of the output map in the affected hemisphere suggested the recruitment of adjacent brain areas. In follow-up examinations up to 6 months after treatment, motor performance remained at a high level, whereas the cortical area sizes in the 2 hemispheres became almost identical, representing a return of the balance of excitability between the 2 hemispheres toward a normal condition. CONCLUSIONS: This is the first demonstration in humans of a long-term alteration in brain function associated with a therapy-induced improvement in the rehabilitation of movement after neurological injury. PMID- 10835435 TI - Warfarin for stroke prevention still underused in atrial fibrillation: patterns of omission. AB - BACKGROUND AND PURPOSE: The value of warfarin in preventing stroke in patients with chronic atrial fibrillation is well established. However, the prevalence of such treatment generally lags behind actual requirements. The aim of this study was to evaluate doctor- and/or patient-related demographic, clinical, and echocardiographic factors that influence decision for warfarin treatment. METHODS: Between 1990 and 1998, 1027 patients were discharged with chronic or persistent atrial fibrillation. This population was composed of (1) patients with cardiac prosthetic valves (n=48), (2) those with increased bleeding risks (n=152), (3) physically or mentally handicapped patients (n=317), and (4) the remaining 510 patients, the main study group who were subjected to thorough statistical analysis for determining factors influencing warfarin use. RESULTS: The respective rates of warfarin use on discharge in the 4 groups were 93.7%, 30.9%, 17.03%, and 59.4% (P=0.001); of the latter, an additional 28.7% were discharged on aspirin. In the main study group, warfarin treatment rates increased with each consecutive triennial period (29.7%, 53.6%, and 77.1%, respectively; P=0.001). Age >80 years, poor command of Hebrew, and being hospitalized in a given medical department emerged as independent variables negatively influencing warfarin use: P=0.0001, OR 0.30 (95% CI 0.17 to 0.55); P=0.02, OR 0.59 (95% CI 0.36 to 0.94); and P=0.0002, OR 0.26 (95% CI 0.12 to 0.52), respectively. In contrast, past history of stroke and availability of echocardiographic information, regardless of the findings, each increased warfarin use (P=0.03, OR 1.95 [95% CI 1.04 to 3.68], and P=0.0001, OR 3.52 [95% CI 2.16 to 5.72], respectively). CONCLUSIONS: Old age, language difficulties, insufficient doctor alertness to warfarin benefit, and patient disability produced reluctance to treat. Warfarin use still lags behind requirements. PMID- 10835436 TI - Medical complications after stroke: a multicenter study. AB - BACKGROUND AND PURPOSE: This prospective, multicenter study was performed to determine the frequency of symptomatic complications up to 30 months after stroke using prespecified definitions of complications. METHODS: We recruited 311 consecutive stroke patients admitted to hospital. Research nurses reviewed their progress on a weekly basis until hospital discharge and again at 6, 18, and 30 months after stroke. RESULTS: Complications during hospital admission were recorded in 265 (85%) of stroke patients. Specific complications were as follows: neurological-recurrent stroke (9% of patients), epileptic seizure (3%); infections-urinary tract infection (24%), chest infection (22%), others (19%); mobility related-falls (25%), falls with serious injury (5%), pressure sores (21%); thromboembolism-deep venous thrombosis (2%), pulmonary embolism (1%); pain shoulder pain (9%), other pain (34%); and psychological-depression (16%), anxiety (14%), emotionalism (12%), and confusion (56%). During follow-up, infections, falls, "blackouts, " pain, and symptoms of depression and anxiety remained common. Complications were observed across all 3 hospital sites, and their frequency was related to patient dependency and duration after stroke. CONCLUSIONS: Our prospective cohort study has confirmed that poststroke complications, particularly infections and falls, are common. However, we have also identified complications relating to pain and cognitive or affective symptoms that are potentially preventable and may previously have been underestimated. PMID- 10835437 TI - Stroke knowledge among stroke support group members. AB - BACKGROUND AND PURPOSE: Community stroke education is needed to improve early stroke recognition and reduce delays in the referral of stroke patients. In some regions, stroke support groups are important promoters of regional stroke education. However, there are no data about the level of stroke knowledge among support group members that support this promotional role. METHODS: We performed a cross-sectional questionnaire survey among 11 German stroke support groups. The questionnaire asked for stroke knowledge and sociodemographic and medical data. Stroke knowledge was excellent if a participant knew (1) at least 2 stroke symptoms (good symptom knowledge) and (2) at least 2 stroke risk factors (good risk factor knowledge), as well as knowing (3) that immediate hospital admission or an emergency call is necessary in case of stroke (good action knowledge). RESULTS: A total of 133 members (96.2%) of 11 stroke support groups took part in the study. Mean age was 65.3 years (SD 11.2 years). Fifty-four percent of subjects were female, 72.8% were retired, and 69.8% were stroke patients. Of the participants, 80.3% had good symptom knowledge, 64.7% had good risk factor knowledge, and 79.7% had good action knowledge. Stroke knowledge was excellent in 44.0% of subjects. Logistic regression analysis showed that age <70 years and not having had a stroke were significant predictors for excellent stroke knowledge. CONCLUSIONS: Overall, members of stroke support groups are well informed about all aspects of modern stroke care. Because of their knowledge and personal experience, support groups should be viewed as important partners in community stroke education. PMID- 10835438 TI - Cervene (Nalmefene) in acute ischemic stroke : final results of a phase III efficacy study. The Cervene Stroke Study Investigators. AB - BACKGROUND AND PURPOSE: The goals of the present study were to assess the efficacy and safety of nalmefene (Cervene) in patients with acute (< or =6 hours) ischemic stroke and to investigate the safety of combined recombinant tissue plasminogen activator and nalmefene in a separate subset of patients. Nalmefene, an opioid antagonist with relative kappa receptor selectivity, has shown neuroprotective effects in multiple experimental central nervous system injury and ischemic models. Results from an earlier phase II study in patients with acute ischemic stroke suggested that nalmefene was safe and tolerable and may be effective for patients <70 years old. METHODS: This investigation was a phase III, placebo-controlled, double-blind, randomized study of a 24-hour infusion of nalmefene. Patients with acute ischemic stroke who had an onset of symptoms within 6 hours and a baseline score of > or =4 on the NIH Stroke Scale were randomized to receive either 60 mg nalmefene administered as a 10-mg bolus over 15 minutes and then a 50-mg infusion over 23.75 hours or placebo. The primary efficacy outcome was the proportion of patients achieving a score of > or =60 on the Barthel Index and a rating of "moderate disability" or better on the Glasgow Outcome Scale at 12 weeks. Assessments were performed at baseline (predose), hours 12 and 24, days 2 and 7, and week 12. RESULTS: A total of 368 patients were randomized at 42 centers, including 32 patients treated with recombinant tissue plasminogen activator and study drug. Nalmefene was well tolerated. Overall, there was no significant difference in 3-month functional outcome for nalmefene treatment compared with placebo on any of the planned analyses. A prospective secondary analysis also failed to find a treatment effect in patients <70 years old. CONCLUSIONS: Although nalmefene appears to be safe and well tolerated, this study failed to find any treatment benefit in stroke patients treated within 6 hours. PMID- 10835439 TI - Indications for early aspirin use in acute ischemic stroke : A combined analysis of 40 000 randomized patients from the chinese acute stroke trial and the international stroke trial. On behalf of the CAST and IST collaborative groups. AB - BACKGROUND AND PURPOSE: Long-term daily aspirin is of benefit in the years after ischemic stroke, and 2 large randomized trials (the Chinese Acute Stroke Trial [CAST] and the International Stroke Trial [IST]), with 20 000 patients in each, have shown that starting daily aspirin promptly in patients with suspected acute ischemic stroke also reduces the immediate risk of further stroke or death in hospital and the overall risk of death or dependency. However, some uncertainty remains about the effects of early aspirin in particular categories of patient with acute stroke. METHODS: To assess the balance of benefits and risks of aspirin in particular categories of patient with acute stroke (eg, the elderly, those without a CT scan, or those with atrial fibrillation), a prospectively planned meta-analysis is presented of the data from 40 000 individual patients from both trials on events that occurred in the hospital during the scheduled treatment period (4 weeks in CAST, 2 weeks in IST), with 10 characteristics used to define 28 subgroups. This represents 99% of the worldwide evidence from randomized trials. RESULTS: There was a highly significant reduction of 7 per 1000 (SD 1) in recurrent ischemic stroke (320 [1.6%] aspirin versus 457 [2. 3%] control, 2P<0.000001) and a less clearly significant reduction of 4 (SD 2) per 1000 in death without further stroke (5.0% versus 5. 4%, 2P=0.05). Against these benefits, there was an increase of 2 (SD 1) per 1000 in hemorrhagic stroke or hemorrhagic transformation of the original infarct (1.0% versus 0.8%, 2P=0.07) and no apparent effect on further stroke of unknown cause (0.9% versus 0.9%). In total, therefore, there was a net decrease of 9 (SD 3) per 1000 in the overall risk of further stroke or death in hospital (8.2% versus 9.1%, 2P=0.001). For the reduction of one third in recurrent ischemic stroke, subgroup-specific analyses found no significant heterogeneity of the proportional benefit of aspirin (chi(2)(18)=20. 9, NS), even though the overall treatment effect (chi(2)(1)=24.8, 2P<0.000001) was sufficiently large for such subgroup analyses to be statistically informative. The absolute risk among control patients was similar in all 28 subgroups, so the absolute reduction of approximately 7 per 1000 in recurrent ischemic stroke does not differ substantially with respect to age, sex, level of consciousness, atrial fibrillation, CT findings, blood pressure, stroke subtype, or concomitant heparin use. There was no good evidence that the apparent decrease of approximately 4 per 1000 in death without further stroke was reversed in any subgroup or that in any subgroup the increase in hemorrhagic stroke was much larger than the overall average of approximately 2 per 1000. Finally, there was no significant heterogeneity between the reductions in the composite outcome of any further stroke or death (chi(2)(18)=16.5, NS). Among the 9000 patients (22%) randomized without a prior CT scan, aspirin appeared to be of net benefit with no unusual excess of hemorrhagic stroke; moreover, even among the 800 (2%) who had inadvertently been randomized after a hemorrhagic stroke, there was no evidence of net hazard (further stroke or death, 63 aspirin versus 67 control). CONCLUSIONS: Early aspirin is of benefit for a wide range of patients, and its prompt use should be routinely considered for all patients with suspected acute ischemic stroke, mainly to reduce the risk of early recurrence. PMID- 10835440 TI - Effect of intravenous nimodipine on blood pressure and outcome after acute stroke. AB - BACKGROUND AND PURPOSE: The Intravenous Nimodipine West European Stroke Trial (INWEST) found a correlation between nimodipine-induced reduction in blood pressure (BP) and an unfavorable outcome in acute stroke. We sought to confirm this correlation with and without adjustment for prognostic variables and to investigate outcome in subgroups with increasing levels of BP reduction. METHODS: Patients with a clinical diagnosis of ischemic stroke (within 24 hours) were consecutively allocated to receive placebo (n=100), 1 mg/h (low-dose) nimodipine (n=101), or 2 mg/h (high-dose) nimodipine (n=94). The correlation between average BP change during the first 2 days and the outcome at day 21 was analyzed. RESULTS: Two hundred sixty-five patients were included in this analysis (n=92, 93, and 80 for placebo, low dose, and high dose, respectively). Nimodipine treatment resulted in a statistically significant reduction in systolic BP (SBP) and diastolic BP (DBP) from baseline compared with placebo during the first few days. In multivariate analysis, a significant correlation between DBP reduction and worsening of the neurological score was found for the high-dose group (beta=0.49, P=0. 048). Patients with a DBP reduction of > or =20% in the high dose group had a significantly increased adjusted OR for the compound outcome variable death or dependency (Barthel Index <60) (n/N=25/26, OR 10. 16, 95% CI 1.02 to 101.74) and death alone (n/N=9/26, OR 4.336, 95% CI 1.131 16.619) compared with all placebo patients (n/N=62/92 and 14/92, respectively). There was no correlation between SBP change and outcome. CONCLUSIONS: DBP, but not SBP, reduction was associated with neurological worsening after the intravenous administration of high-dose nimodipine after acute stroke. For low-dose nimodipine, the results were not conclusive. These results do not confirm or exclude a neuroprotective property of nimodipine. PMID- 10835441 TI - Urokinase cisternal irrigation therapy for prevention of symptomatic vasospasm after aneurysmal subarachnoid hemorrhage: a study of urokinase concentration and the fibrinolytic system. AB - BACKGROUND AND PURPOSE: Cisternal irrigation therapy with urokinase (UK) was performed in multiple institutions to prevent symptomatic vasospasm. The efficacy and safety of this therapy were evaluated, and the optimal concentration of UK was estimated. METHODS: This therapy was performed in 28 patients who underwent surgery within 72 hours of the onset of severe subarachnoid hemorrhage (Fisher's group 3, CT number [Hounsfield units] >60). After the aneurysm was clipped, irrigation tubes were placed in the Sylvian fissure (inlet) unilaterally and in the prepontine or chiasmatic cistern (outlet). Lactated Ringer's solution with UK (30, 60, or 120 IU/mL) was infused at a rate of 30 mL/h. The presence of symptomatic vasospasm was evaluated by changes in the clinical symptoms and the presence of a new low-density area on CT scan. Drained irrigation fluid and peripheral blood were examined chronologically to evaluate the fibrinolytic system. RESULTS: Symptomatic vasospasm was observed transiently in 3 cases (10.7%) without any low-density area on CT scan. In the 120-IU/mL group, no symptomatic vasospasm occurred. Analysis of drainage fluid suggested that UK 120 IU/mL is effective. The mean values of total drained blood volume for the respective groups were as follows: 58 mL in 30 IU/mL, 106 mL in 60 IU/mL, and 143 mL in 120 IU/mL. No abnormal changes were observed in the coagulative and fibrinolytic systems after UK irrigation. CONCLUSIONS: These results suggest that cisternal irrigation therapy with UK is safe and effective for the prevention of symptomatic vasospasm after aneurysmal subarachnoid hemorrhage. PMID- 10835442 TI - Effect of the apolipoprotein E epsilon4 allele on white matter hyperintensities in dementia. AB - BACKGROUND AND PURPOSE: The clinical significance of the apoE epsilon4 allele in white matter changes in patients with dementia has been a subject of debate. We studied the association between the apoE epsilon4 allele and white matter hyperintensities (WMHs) before and after control for (1) potential vascular risk factors and (2) the presence of lacunar infarcts in patients with dementia. METHODS: The subjects were 131 patients with dementia who had either Alzheimer's disease or vascular dementia, or a combination of these 2 types of dementia, with or without WMHs, lacunar infarcts, or both. The association of the epsilon4 allele with WMHs was examined before and after control for age, sex, duration of symptoms, education level, severity of dementia, presence of lacunar infarcts, and potential vascular risk factors, including hypertension, diabetes mellitus, lipid disorders, smoking habit, drinking habit, and cardiac diseases. RESULTS: WMHs were observed in 73 (55.7%) of the patients. Neither the number of apoE epsilon4 alleles nor their presence was significantly associated with WMHs before or after control for the potential confounding factors. Multiple logistic regression analyses revealed that age, the presence of hypertension, and the presence of lacunar infarcts were independently associated with WMHs. CONCLUSIONS: The apoE epsilon4 allele was not associated with WMHs in patients with dementia. The fact that WMHs were significantly associated with hypertension and lacunar infarcts may indicate an ischemic origin of WMHs. PMID- 10835443 TI - The effect of acute ingestion of a large dose of alcohol on the hemostatic system and its circadian variation. AB - BACKGROUND AND PURPOSE: Heavy binge drinking may trigger the onset of embolic stroke and acute myocardial infarction, but the underlying mechanisms are unclear. The effects of binge drinking on the hemostatic system and its circadian variation have not been investigated. We investigated the effects of an acute intake of a large dose of alcohol (1.5 g/kg). METHODS: Twelve healthy, nonsmoking men participated in sessions where they were served ethanol in fruit juice or served fruit juice alone and, lying in a supine position, were followed up for 12 to 24 hours. The treatments were randomized and separated from each other by a 1 week washout period. Blood and urine were collected for hemostatic measurements. RESULTS: The urinary excretion of the platelet thromboxane A(2) metabolite 2, 3 dinor-thromboxane B(2) was significantly (P<0.05) greater during the night after an evening intake of alcohol than during the control night. A smaller increase was observed during the daytime after an intake of alcohol in the morning. The effects on the endothelial prostacyclin metabolite 2,3-dinor-6-ketoprostaglandin F(1alpha) excretion were negligible. A 7-fold increase in plasminogen activator inhibitor 1 activity was observed after both morning (P<0. 05) and evening (P<0.01) intakes of alcohol. CONCLUSIONS: This is the first study to suggest that acute ingestion of a relatively large but tolerable dose of alcohol transiently enhances thromboxane-mediated platelet activation. The observations also demonstrate alcohol-induced changes in the normal circadian periodicity of the hemostatic system in subjects not accustomed to consumption of alcohol. PMID- 10835444 TI - Risk factors for peripartum and postpartum stroke and intracranial venous thrombosis. AB - BACKGROUND AND PURPOSE: The study goal was to identify potential risk factors for peripartum or postpartum stroke and intracranial venous thrombosis. METHODS: Data from the Healthcare Cost and Utilization Project were analyzed for the years 1993 and 1994. Observed values were weighted with poststratification discharge weights to project to the universe of all discharges from community hospitals located in the United States. Nationally representative estimates of risk were calculated on the basis of age, race, mode of delivery, income, third-party payer, hospital size, hospital ownership, hospital location (rural versus urban), hospital teaching status, census region, and presence of specific complications. Multivariate models were developed with the use of logistic regression. RESULTS: Among 1 408 015 sampled deliveries, there were 183 observed cases of peripartum stroke and 170 cases of peripartum intracranial venous thrombosis in 17 states in the United States in 1993 and 1994. There were an estimated 975 cases of stroke and 864 cases of intracranial venous thrombosis during pregnancy and the puerperium in the United States among 7 463 712 deliveries during 1993 and 1994, for estimated risks of 13.1 cases of peripartum stroke and 11.6 cases of peripartum intracranial venous thrombosis per 100 000 deliveries. Multivariate analysis showed that the following were strongly and significantly associated with both peripartum and postpartum stroke: cesarean delivery; fluid, electrolyte, and acid-base disorders; and hypertension. Covariates that were strongly and significantly associated with both peripartum and postpartum intracranial venous thrombosis included cesarean delivery, hypertension, and infections other than pneumonia and influenza. CONCLUSIONS: Pregnancy-related hypertension and cesarean delivery are important risk factors for both stroke or intracranial venous thrombosis. PMID- 10835445 TI - Factor V Leiden and antiphospholipid antibodies are significant risk factors for ischemic stroke in children. AB - BACKGROUND AND PURPOSE: The association between ischemic childhood stroke and thrombophilia has been debated. We studied the prevalence of thrombophilia risk factors in 65 unrelated children with ischemic stroke compared with 145 control subjects. METHODS: Patients and control subjects were tested for antithrombin protein C and protein S deficiencies, the presence of antiphospholipid antibodies (APLA), factor V Leiden (FVL), G20210A polymorphism of factor II gene (FII G20210A), and C677T polymorphism of 5,10-methylenetetrahydrofolate reductase gene (C677T MTHFR). RESULTS: Of 65 children, 7 had a stroke in the neonatal/perinatal period and therefore were analyzed separately. Thirty-one of the remaining 58 patients with pediatric stroke (53.4%) were found to have at least 1 thrombophilia marker compared with only 25.5% of control subjects. None of the patients or control subjects had protein S or antithrombin III deficiency. The prevalence of protein C deficiency was higher among pediatric stroke patients than among control subjects, but the difference was not statistically significant (OR=7, 95% CI 0.75 to 65.1). Heterozygous FII G20210A and homozygous MTHFR 677T were not associated with an increased risk for stroke (OR=1.29, 95% CI 0.2 to 8.2; and OR=1.06, 95% CI 0.4 to 2.7, respectively). In contrast, the presence of APLA was associated with a >6-fold risk of stroke (OR=6. 08, 95% CI 1.5 to 24.3), and the heterozygosity for FVL increased the risk of stroke by almost 5-fold (OR=4.82, 95% CI 1.4 to 16.5). Five patients with pediatric stroke had a combination of > or =2 thrombophilia markers, whereas none of the control subjects had a combination of the markers. Most of the patients with neonatal/perinatal stroke were found to have at least 1 thrombophilia marker. CONCLUSIONS: These data suggest that the prevalence of thrombophilia markers is increased in children with stroke compared with control subjects and, specifically, that FVL and APLA contribute significantly to stroke occurrence. PMID- 10835446 TI - Anticardiolipin antibodies are not an independent risk factor for stroke: an incident case-referent study nested within the MONICA and Vasterbotten cohort project. AB - BACKGROUND AND PURPOSE: Anticardiolipin antibodies (aCL) have been proposed to be an independent risk factor for stroke. To test this hypothesis, a nested case control study was performed to compare aCL with the other known risk factors for stroke. METHODS: Within the framework of the World Health Organization Multinational Monitoring of Trends and Determinants in Cardiovascular Disease (MONICA) project and the Vasterbotten Intervention Program (VIP) health survey, 44 725 men and women were enrolled and followed up from January 1, 1985, through August 31, 1996. Individuals free from cardiovascular events were followed up, and 123 developed stroke (on average, 34.1 months after blood sampling; 21 cerebral hemorrhage and 102 cerebral infarction); they were compared with 241 age and sex-matched control subjects from the same population. ELISA was used for the analysis of IgG, IgM, and IgA aCL. RESULTS: IgM-aCL were present in 11.4% of patients (14/123) who developed stroke and in 4. 1% of individuals (10/241) who remained healthy (P=0.013, OR 2.97, 95% CI 1.28 to 6.89). The OR for the levels of IgM-aCL was 1.34 (P=0. 01, 95% CI 1.07 to 1.68) without adjustment for other risk factors and 1.24 when adjusted for hypertension, diabetes mellitus, cigarette smoking, and use of smokeless tobacco (P=0.077, 95% CI 0. 98 to 1.56). There was no difference between patients and controls for the prevalence or level of IgG-aCL and IgA-aCL and also no difference between patients with cerebral hemorrhage and cerebral infarction for the prevalence of all 3 isotypes of aCL. CONCLUSIONS: We conclude that aCL are associated with future stroke but do not constitute an independent risk factor. PMID- 10835447 TI - Ischemic heart disease in black South African stroke patients. AB - BACKGROUND AND PURPOSE: Stroke patients in western countries frequently have coronary artery disease (CAD). In black Africans, CAD has been reported as being rare in both stroke patients and the general population. In this study, an attempt has been made to determine the prevalence of CAD in a black South African stroke population. METHODS: The prevalence of CAD was determined by indicators identified through a series of 5 observational studies in black patients diagnosed with stroke. CAD indicators included (1) bedside diagnosis in 741 patients; (2) resting ECG in 555 consecutively admitted patients; (3) a combination of clinical examination, cardiac ultrasound, radionuclide scintigraphy, and multigated blood pool studies in 102 consecutively admitted patients; (4) thallium scintigraphy in 60 patients; and (5) necropsy in 23 patients. RESULTS: On bedside questioning, only 0.7% complained of previous angina. There was no history given of myocardial infarction (MI), but documentation of this was found in the clinical notes of 0.7% of the patients. In the resting ECG study, evidence of myocardial ischemia was present in 14.6% and MI in 2.1%. In the combined study, cardiac ischemia was documented on ECG in 12.7% of patients and evidence of previous MI in 5.8%. Cardiac scintigraphic studies revealed changes of myocardial ischemia in 31.7% and MI in 13.3% of the 60 patients studied. Four (17.4%) of 23 patients in the necropsy study had histological evidence of previous MI, and 50% of all patients had evidence of >50% atherosclerotic stenosis in 1, 2, or 3 coronary arteries. CONCLUSIONS: The prevalence of CAD in black African stroke patients is significantly higher than has been documented in the general nonstroke black population as well as in stroke patients. Black stroke patients may have a risk for CAD similar to that of their white counterparts. PMID- 10835448 TI - Age-dependent association of apolipoprotein E genotypes with stroke subtypes in a Japanese rural population. AB - BACKGROUND AND PURPOSE: The association between apolipoprotein E (apoE) polymorphisms and stroke has been controversial. These controversies may be due to inaccurate classification of stroke and differences in age ranges. We investigated the association between apoE genotypes and stroke subtypes (confirmed by CT or MRI findings) by case-control study in a Japanese rural population. METHODS: First-ever-stroke patients (n=322; cerebral infarction, n=201, intracerebral hemorrhage, n=84, and subarachnoid hemorrhage, n=37) aged 40 to 89 years were recruited from Hokuetsu Hospital, Japan. Healthy controls (n=1126) were selected from the general population in the same area. ApoE genotypes were determined by restriction fragment-length polymorphism analysis. RESULTS: Compared with apoE epsilon3/epsilon3 subjects, epsilon2 carriers had a 2 fold risk of cerebral infarction (OR 1.9, 95% CI 1.1 to 3.2). Among cerebral infarction patients, epsilon2 carriers had increased risks of cortical infarction (OR 2.4, 95% CI 1.3 to 4.6) (an anatomic subtype) and atherothrombosis (OR 3.9, 95% CI 1.7 to 9.0) and cardioembolism (OR 4.9, 95% CI 1.6 to 14.4) but not lacunar infarction (clinical subtypes). ApoE epsilon4 carriers had a 2. 5-fold risk of subarachnoid hemorrhage (OR 2.5, 95% CI 1.1 to 5.4). ApoE epsilon2/epsilon2 subjects had an increased risk of intracerebral hemorrhage (OR 4.4, 95% CI 1.0 to 19.7). ApoE epsilon3/epsilon4 subjects showed approximately 2 fold increased risk of atherothrombosis (OR 2.1, 95% CI 1.0 to 4.1) and intracerebral hemorrhage (OR 1.8, 95% CI 1.0 to 3.3). The association between epsilon2 and stroke was accentuated in subjects aged 70 years or older but not in those aged 40 to 69 years. CONCLUSIONS: Our study suggests that apoE epsilon2 is a risk factor for atherothrombosis, cardioembolism, and intracerebral hemorrhage, whereas epsilon4 is a risk factor for atherothrombosis, intracerebral hemorrhage, and subarachnoid hemorrhage. The occurrence of stroke may be affected by interaction between age and apoE gene polymorphisms. PMID- 10835449 TI - Demographic, morphological, and clinical characteristics of 1289 patients with brain arteriovenous malformation. AB - BACKGROUND AND PURPOSE: The purpose of this study was to assess demographic, clinical, and morphological characteristics of patients with brain arteriovenous malformations (AVMs). METHODS: Prospectively collected data of 1289 consecutive AVM patients from 3 independent databases (1 multicenter [Berlin/Paris/Middle and Far East, n=662] and 2 single centers [New York, n=337, and Toronto, n=290]) were analyzed. The variables assessed were age at diagnosis, sex, AVM size, AVM drainage pattern, AVM location in functionally important brain areas ("eloquence"), and type of presentation (hemorrhage, seizure, chronic headache, or focal neurologic deficit). Comparisons were made by ANOVA, contingency tables, and log-linear models. RESULTS: Overall, mean age at diagnosis was 31.2 years (95% CI 30.2 to 32.2 years), and 45% of the patients were female (95% CI 42% to 47%). AVM maximum diameter was <3 cm in 38% (95% CI 35% to 41%). Deep venous drainage was present in 55% (95% CI 52% to 59%). An eloquent AVM location was described in 71% (95% CI 69% to 74%). AVM hemorrhage occurred in 53% (95% CI 51% to 56%). Generalized or focal seizures were described in 30% (95% CI 27% to 33%) and 10% (95% CI 8% to 12%), respectively. Chronic headache was recorded in 14% (95% CI 12% to 16%). Persistent neurological deficits were found in 7% (95% CI 6% to 9%), and progressive neurological deficits in 5% (95% CI 4% to 6%). Significant differences between centers were found for age (P<0.001), sex (P=0.04), eloquence (P=0.04), size (P<0.001), hemorrhage (P=0.006), persistent neurological deficit (P<0.001), and reversible neurological deficit (P=0.013). The intercenter difference found for hemorrhage frequency did not remain after adjustment for AVM size. CONCLUSIONS: Baseline characteristics differed considerably between centers. The differences found in patient age and AVM size may be explained by center-specific referral patterns and the influence of access to treatment resources, whereas those found for other characteristics may be attributable to center-specific definitions. Analysis of natural history data from tertiary referral center databases may be improved by consistent definitions applicable to the entire population of AVM patients. PMID- 10835450 TI - Diffusion- and perfusion-weighted MRI: influence of severe carotid artery stenosis on the DWI/PWI mismatch in acute stroke. AB - BACKGROUND AND PURPOSE: Diffusion-weighted imaging (DWI) and perfusion-weighted imaging (PWI) have been used increasingly in recent years to evaluate acute stroke in the emergency setting. In the present study, we compared DWI and PWI findings in acute stroke patients with and without severe extracranial internal carotid artery (ICA) disease. METHODS: Twenty-seven patients with nonlacunar ischemic stroke were selected for this analysis. DWI, PWI, and conventional MRI were performed in all patients within 24 hours of symptom onset and after 1 week. To exclude patients with partial or complete reperfusion, we included only patients with a PWI deficit larger than the DWI lesion. Severe ICA disease (>70% stenosis) was present unilaterally in 9 and bilaterally in 2 patients. Acute DWI lesion volume, the size of the acute PWI/DWI mismatch, and final infarct size (on T2-weighted images) were determined. RESULTS: The PWI/DWI mismatch was significantly larger in patients with severe ICA disease than in patients without extracranial carotid stenosis, both when time-to-peak and mean transit time maps (P<0.01) were used to calculate the mismatch. Quantitative analysis of the time to-peak delay in the mismatch indicated that a relatively smaller fraction of the total mismatch was critically ischemic in patients with carotid stenosis than in those without. Average lesion volume increased less in the stenosis group (P=0.14), despite the larger PWI/DWI mismatch, and final infarct size was smaller in the stenosis group (P<0.05). In the 2 patients with bilateral ICA disease, variable hemodynamic involvement of the contralateral hemisphere was found in addition to the ipsilateral PWI deficit. CONCLUSIONS: In most acute stroke patients with severe ICA stenosis, a considerably smaller fraction of the total PWI/DWI mismatch is at risk than in patients without carotid disease. PMID- 10835451 TI - Monitoring intravenous recombinant tissue plasminogen activator thrombolysis for acute ischemic stroke with diffusion and perfusion MRI. AB - BACKGROUND AND PURPOSE: Intravenous recombinant tissue plasminogen activator (rtPA) administration is an effective therapy for ischemic stroke when initiated within 3 hours and possibly up to 6 hours after symptom onset. To improve patient selection, a fast diagnostic tool that allows reliable diagnosis of hemorrhage and ischemia, vessel status, and tissue at risk at an early stage may be useful. We studied the feasibility of stroke MRI for the initial evaluation and follow-up monitoring of patients undergoing intravenous thrombolysis. METHODS: Stroke MRI (diffusion- and perfusion-weighted imaging [DWI and PWI, respectively], magnetic resonance angiography, and T2-weighted imaging) was performed before, during, or after thrombolysis and on days 2 and 5. We assessed clinical scores (National Institutes of Health Stroke Scale [NIHSS], Scandinavian Stroke Scale [SSS], Barthel Index, and Rankin scale) at days 1, 2, 5, 30, and 90. Furthermore, we performed volumetric analysis of infarct volumes on days 1, 2, and 5 as shown in PWI, DWI, and T2-weighted imaging. RESULTS: Twenty-four patients received rtPA within a mean time interval after symptom onset of 3.27 hours and stroke MRI of 3.43 hours. Vessel occlusion was present in 20 of 24 patients; 11 vessels recanalized (group 1), and 9 did not (group 2). The baseline PWI lesion volume was significantly larger (P=0.008) than outcome lesion size in group 1, whereas baseline DWI lesion volume was significantly smaller (P=0.008) than final infarct size in group 2. Intergroup outcome differed significantly for all scores at days 30 and 90 (all P<0.01). Intragroup differences were significant in group 1 for change in SSS and NIHSS between day 1 and day 30 (P=0.003) and for SSS only between day 1 and day 90 (P=0.004). CONCLUSIONS: Stroke MRI provides comprehensive prognostically relevant information regarding the brain in hyperacute stroke. Stroke MRI may be used as a single imaging tool in acute stroke to identify and monitor candidates for thrombolysis. It is proposed that stroke MRI is safe, reliable, and cost effective; however, our data do not prove this assumption. Early recanalization achieved by thrombolysis can save tissue at risk if present and may result in significantly smaller infarcts and a significantly better outcome. PMID- 10835452 TI - Comparison of microembolism detected by transcranial Doppler and neuropsychological sequelae of carotid surgery and percutaneous transluminal angioplasty. AB - BACKGROUND AND PURPOSE: Percutaneous transluminal angioplasty (PTA) is currently being assessed for the treatment of carotid stenosis. In comparison with carotid endarterectomy (CEA), there is evidence of an increased risk of cerebral microembolism during the procedure. We have sought evidence of any neuropsychological sequelae of carotid PTA and compared it with CEA to demonstrate the relative safety of the 2 treatment options. METHODS: The neuropsychological outcomes after CEA and PTA were compared in 2 matched groups of patients with severe symptomatic carotid stenosis, 96% of whom had been randomized in the Carotid and Vertebral Artery Transluminal Angioplasty Study (CAVATAS), at a single center. Transcranial Doppler insonation of the middle cerebral artery was used to measure cerebral reactivity in response to carbon dioxide inhalation before treatment and then to detect microembolization of the ipsilateral cerebral hemisphere and measure changes in blood flow velocity during the procedures. The performance on a neuropsychological test battery administered before, 6 weeks after, and 6 months after the procedure was compared in 20 patients undergoing PTA and 26 having CEA. RESULTS: At 6 weeks, 5 patients in each group showed a similar decline in neuropsychological performance; global measures showed no significant difference between the 2 procedures, despite a significantly higher incidence of microemboli during PTA. Both groups showed a marked reduction in anxiety after treatment. CONCLUSIONS: The findings provide some reassurance that PTA is not associated with greater cerebral complications than CEA, despite the higher embolic load recorded by transcranial Doppler ultrasonography during angioplasty. PMID- 10835453 TI - Evaluation of new online automated embolic signal detection algorithm, including comparison with panel of international experts. AB - BACKGROUND AND PURPOSE: The clinical application of Doppler detection of circulating cerebral emboli will depend on a reliable automated system of embolic signal detection; such a system is not currently available. Previous studies have shown that frequency filtering increases the ratio of embolic signal to background signal intensity and that the incorporation of such an approach into an offline automated detection system markedly improved performance. In this study, we evaluated an online version of the system. In a single-center study, we compared its performance with that of a human expert on data from 2 clinical situations, carotid stenosis and the period immediately after carotid endarterectomy. Because the human expert is currently the "gold standard" for embolic signal detection, we also compared the performance of the system with an international panel of human experts in a multicenter study. METHODS: In the single-center evaluation, the performance of the software was tested against that of a human expert on 20 hours of data from 21 patients with carotid stenosis and 18 hours of data from 9 patients that was recorded after carotid endarterectomy. For the multicenter evaluation, a separate 2-hour data set, recorded from 5 patients after carotid endarterectomy, was analyzed by 6 different human experts using the same equipment and by the software. Agreement was assessed by determining the probability of agreement. RESULTS: In the 20 hours of carotid stenosis data, there were 140 embolic signals with an intensity of > or =7 dB. With the software set at a confidence threshold of 60%, a sensitivity of 85.7% and a specificity of 88.9% for detection of embolic signals were obtained. At higher confidence thresholds, a specificity >95% could be obtained, but this was at the expense of a lower sensitivity. In the 18 hours of post-carotid endarterectomy data, there were 411 embolic signals of > or =7-dB intensity. When the same confidence threshold was used, a sensitivity of 95.4% and a specificity of 97.5% were obtained. In the multicenter evaluation, a total of 127 events were recorded as embolic signals by at least 1 center. The total number of embolic signals detected by the 6 different centers was 84, 93, 108, 92, 63, and 78. The software set at a confidence threshold of 60% detected 90 events as embolic signals. The mean probability of agreement, including all human experts and the software, was 0.83, and this was higher than that for 2 human experts and lower than that for 4 human experts. The mean values for the 6 human observers were averaged to give P=0.84, which was similar to that of the software. CONCLUSIONS: By using the frequency specificity of the intensity increase occurring with embolic signals, we have developed an automated detection system with a much improved sensitivity. Its performance was equal to that of some human experts and only slightly below the mean performance of a panel of human experts PMID- 10835454 TI - Preferred technique for blood flow volume measurement in cerebrovascular disease. AB - BACKGROUND AND PURPOSE: A noninvasive reliable technique that can reveal cerebral blood flow volume could be a valuable tool in screening programs for stroke prevention. In diagnostic ultrasonography, spectral Doppler imaging (SDI) is popular among sonologists and vascular technologists to estimate blood flow volume despite its documented inaccuracy and the availability of the more accurate technique of color velocity imaging (CVI). The aim of the present study was to demonstrate the discrepancy of blood flow volume estimation with CVI and SDI with use of an "internal" standard. METHODS: The common, internal, and external carotid arteries of 50 healthy subjects (22 men, 28 women, age range 19 to 54 years) were examined with CVI and SDI. The total blood flow volume of the internal and external carotid arteries was then compared with the ipsilateral common carotid artery flow. An accurate technique would demonstrate no difference. The difference (expressed as a percent inconsistency) was therefore a measure of the accuracy of the method. RESULTS: The mean+/-SD inconsistency was found to be 10.6+/-8.3% for CVI and 27.9+/-14.3% for SDI. The difference in inconsistency between CVI and SDI in measurement of carotid blood flow volume was statistically significant (P<0.01). CONCLUSIONS: CVI is more accurate than SDI in the determination of blood flow volume in the carotid arteries. For noninvasive clinical estimation of cerebrovascular blood flow volume, CVI quantification should be the preferred technique. PMID- 10835455 TI - Collateral configuration of the circle of Willis: transcranial color-coded duplex ultrasonography and comparison with postmortem anatomy. AB - BACKGROUND AND PURPOSE: The anterior communicating artery (AcoA) and posterior communicating arteries (PcoA) of the circle of Willis provide the main route for collateral blood flow in cases of carotid artery obstruction. Transcranial color coded duplex ultrasonography (TCCD) allows real-time measurement of the collateral function of the AcoA and PcoA. The primary objective of this study was to determine the collateral artery threshold diameters for supplying collateral flow. METHODS: In 12 acute stroke patients with a median age of 75 years (51 to 91 years), the collateral integrity of the circle of Willis as assessed by TCCD and carotid compression tests was compared with their postmortem anatomy. The lengths and diameters of the collateral arteries were measured. RESULTS: TCCD demonstrated absent anterior collateral flow in 3 patients. In 1 of these patients, absence of anterior cross-flow was due to an occluded anterior cerebral artery, which was revealed at autopsy. Absent posterior collateral flow was found in 14 hemispheres. In 2 of these hemispheres, autopsy revealed a fetal configuration of the posterior cerebral artery hampering posterior collateral flow. The median (range) diameters as found at autopsy of the functional (n=19) and nonfunctional (n=16) collateral arteries of the circle of Willis were 1.1 (0.4 to 2.0) and 0.5 (0.3 to 0.7) mm, respectively (P=0.003). PcoA diameters were found to correlate negatively (rho=-0. 50, P=0.01) to the diameters of their accessory P1 segments. CONCLUSIONS: The threshold diameter allowing for cross flow through the primary collateral arteries of the circle of Willis is between 0. 4 and 0.6 mm. PMID- 10835456 TI - Stroke patients in south Madrid: function and motor recovery, resource utilization, and family support. AB - BACKGROUND AND PURPOSE: The purpose of the present study was to describe the epidemiology of stroke disability and the use of health resources in South Madrid. METHODS: Among a population of 665 168 residents in South Madrid, patients with an acute stroke of clinical onset during March to July 1996 who were seen at a general hospital or at 1 of 3 primary care centers were evaluated at baseline (n=147) and at 3 months (n=110) and 6 months (n=112) after stroke. We assessed the frequencies of stroke and stroke-related residual disability per population unit, as well as the impairment, disability, secondary complications, use of health resources, and impact on quality of life. RESULTS: In patients > or =60 years old, the incidence of stroke with severe residual disability after 6 months was 75 per 10 000, was higher in men, and increased with age; the proportion of survivors among those examined at baseline was 20%. The use of hospital days per population unit was similar to that of reported European data, but the use of other health care resources was less. Patients frequently used bladder and nasal catheters and presented with shoulder pain. Social activities were infrequent and decreased after stroke. Access to technical aids was limited, and home adaptations were exceptional. The impact of stroke on health-related quality of life among patients and main caregivers was modest. CONCLUSIONS: The study shows that in South Madrid, (1) the use of health resources after stroke is low; (2) patients with stroke register low activities of daily living scores with a comparatively small impact on quality of life; and (3) relative to need, the use of rehabilitation, aids, and home adaptations and services was low. PMID- 10835457 TI - Chronic motor dysfunction after stroke: recovering wrist and finger extension by electromyography-triggered neuromuscular stimulation. AB - BACKGROUND AND PURPOSE: After stroke, many individuals have chronic unilateral motor dysfunction in the upper extremity that severely limits their functional movement control. The purpose of this study was to determine the effect of electromyography-triggered neuromuscular electrical stimulation on the wrist and finger extension muscles in individuals who had a stroke > or = 1 year earlier. METHODS: Eleven individuals volunteered to participate and were randomly assigned to either the electromyography-triggered neuromuscular stimulation experimental group (7 subjects) or the control group (4 subjects). After completing a pretest involving 5 motor capability tests, the poststroke subjects completed 12 treatment sessions (30 minutes each) according to group assignments. Once the control subjects completed 12 sessions attempting wrist and finger extension without any external assistance and were posttested, they were then given 12 sessions of the rehabilitation treatment. RESULTS: The Box and Block test and the force-generation task (sustained muscular contraction) revealed significant findings (P<0. 05). The experimental group moved significantly more blocks and displayed a higher isometric force impulse after the rehabilitation treatment. CONCLUSIONS: Two lines of evidence clearly support the use of the electromyography-triggered neuromuscular electrical stimulation treatment to rehabilitate wrist and finger extension movements of hemiparetic individuals > or =1 year after stroke. The treatment program decreased motor dysfunction and improved the motor capabilities in this group of poststroke individuals. PMID- 10835458 TI - Improved functional outcome in patients with hemorrhagic stroke in putamen and thalamus compared with those with stroke restricted to the putamen or thalamus. AB - BACKGROUND AND PURPOSE: We analyzed the effect of late intensive inpatient rehabilitation on the functional outcome of patients with subcortical hemorrhagic stroke. METHODS: Patients who were nonambulatory with hemorrhagic stroke in the internal capsule and putamen (n=55), the thalamus (n=24), or all 3 regions (n=15) underwent intensive inpatient rehabilitation. Patients with surgical intervention or an episode of ventricular hemorrhage were excluded. Lesion location was evaluated by MRI 4 months after the ictus. RESULTS: Demographic data, initial disability, and impairment measures were comparable in the 3 groups. Functional outcome demonstrated significant differences in mobility subscores (P<0.05) of the Functional Independence Measure such that patients with injury in the 3 regions were more likely to ambulate independently than were patients in the other groups. Lesion location data demonstrated that the ventral anterior nucleus of the thalamus was always spared; the ventral posterior (lateral and medial) nucleus was always damaged, and the ventral lateral nucleus was frequently damaged. Putaminal damage always included the postcommissural area. In addition, the entire posterior half limb of the internal capsule was always damaged. CONCLUSIONS: Subcortical lesions to multiple structures in the basal ganglia thalamocortical motor circuits permitted enhanced motor recovery. Lesion location predicted the level of independent ambulation and the rate of recovery in patients with stroke who were nonambulatory before neurorehabilitation therapy. PMID- 10835459 TI - Mechanisms of dysphagia in suprabulbar palsy with lacunar infarct. AB - BACKGROUND AND PURPOSE: The objective of the present study was to investigate the neural mechanisms of dysphagia in suprabulbar palsy (SBP) with multiple lacunar infarct. METHODS: We evaluated the swallowing disorders of patients with SBP (n=34) and age-matched healthy control subjects (n=35) by means of an electrophysiological method that recorded the oropharyngeal swallowing patterns. With this method, dysphagia limit, the triggering of voluntarily initiated swallows, duration of laryngeal relocation time, and total duration of oropharyngeal swallowing were recorded and measured. In addition, the EMG behavior of the cricopharyngeal (CP) muscle of the upper esophageal sphincter was also assessed. RESULTS: In patients with SBP, the dysphagia limit in all except 1 patient was pathological with limits of <20-mL bolus volume, which is contrary to normal subjects, in whom the dysphagia limit exceeds the 20-mL bolus volume. Either triggering of swallowing reflex was delayed (P<0.04), or the swallow could hardly be triggered in 7 patients on the voluntary attempts for 3 mL water. Whenever the reflex swallowing could be triggered, it was slow and prolonged (P<0.01). The CP muscle of the upper esophageal sphincter appeared to have become hyperreflexic and incoordinated with laryngeal movements during swallowing. CONCLUSIONS: It was proposed that the progressive involvement of the excitatory and inhibitory corticobulbar fiber systems linked with the bulbar swallowing center is mainly responsible for the triggering difficulties of the swallowing reflex and for the hyperreflexic/incoordinated nature of the CP sphincter. In addition, the dysfunction of the extrapyramidal system has a specific role in the slowing of oropharyngeal swallowing and the accumulation of saliva in the mouth. PMID- 10835460 TI - Mechanism of ATP-induced [Ca(2+)](i) mobilization in rat basilar smooth muscle cells. AB - BACKGROUND AND PURPOSE: We have previously reported that extracellular ATP activates P(2u) receptors and increases intracellular free Ca(2+) ([Ca(2+)](i)) by G protein/phospholipase C/inositol 1,4,5-triphosphate pathways in cerebral artery smooth muscle cells. However, the possible contribution of other signaling pathways remains unclear. This study was undertaken to investigate the role of protein tyrosine kinase (PTK) and mitogen-activated protein kinase (MAPK) in mediating ATP-induced Ca(2+) mobilization in rat basilar artery smooth muscle cells (RBASMCs). METHODS: RBASMCs were freshly isolated, and [Ca(2+)](i) was monitored by fura 2 microfluorimetry. MAPK phosphorylation was studied by the Western blot technique. RESULTS: ATP produced a biphasic [Ca(2+)](i) response, which consists of releasing Ca(2+) from internal stores and influx from extracellular space. PTK inhibitors tyrphostin 51 and genistein inhibited [Ca(2+)](i) response to ATP. Tyrphostin A1, an inactive analogue of tyrphostins, failed to reduce the ATP-induced response. MAPK kinase inhibitor PD98059, but not U0126, reduced the ATP-induced [Ca(2+)](i) response. Phosphatidylinositol 3 kinase (PI3-K) tyrosine kinase inhibitor wortmannin, but not janus tyrosine kinase (JAK2) inhibitor AG490, partially inhibited the [Ca(2+)](i) response induced by ATP. In addition, ATP enhanced MAPK phosphorylation in a concentration and time-dependent manner, and genistein, tyrphostin 51, PD98059, and U0126 inhibited MAPK phosphorylation. CONCLUSIONS: Extracellular ATP produced [Ca(2+)](i) elevation and MAPK phosphorylation in RBASMCs, and the effect was regulated by PTK. The role of MAPK in ATP-induced [Ca(2+)](i) elevation is not clear. PI3-K tyrosine kinase and JAK2 tyrosine kinase may not play an important role in the ATP-induced [Ca(2+)](i) response in RBASMCs. PMID- 10835462 TI - Mild and moderate hypothermia (alpha-stat) do not impair the coupling between local cerebral blood flow and metabolism in rats. AB - BACKGROUND AND PURPOSE: The effects of hypothermia on global cerebral blood flow (CBF) and glucose utilization (CGU) have been extensively studied, but less information exists on a local cerebral level. We investigated the effects of normothermic and hypothermic anesthesia on local CBF (LCBF) and local CGU (LCGU). METHODS: Thirty-six rats were anesthetized with isoflurane (1 MAC) and artificially ventilated to maintain normal PaCO(2) (alpha-stat). Pericranial temperature was maintained normothermic (37.5 degrees C, n=12) or was reduced to 35 degrees C (n=12) or 32 degrees C (n=12). Pericranial temperature was maintained constant for 60 min until LCBF and LCGU were measured with autoradiography. Twelve conscious rats served as normothermic control animals. RESULTS: Normothermic anesthesia significantly increased mean CBF compared with conscious control animals (29%, P<0.05). Mean CBF was reduced to control values with mild hypothermia and to 30% below control animals with moderate hypothermia (P<0.05). Normothermic anesthesia reduced mean CGU by 44%. No additional effects were observed during mild hypothermia. Moderate hypothermia resulted in a further reduction in mean CGU (41%, P<0.05). Local analysis showed linear relationships between LCBF and LCGU in normothermic conscious (r=0.93), anesthetized (r=0.92), and both hypothermic groups (35 degrees C r=0. 96, 32 degrees C r=0.96, P<0.05). The LCBF-to-LCGU ratio increased from 1.5 to 2.5 mL/micromol during anesthesia (P<0.05), remained at 2.4 mL/micromol during mild hypothermia, and decreased during moderate hypothermia (2.1 mL/micromol, P<0.05). CONCLUSIONS: Anesthesia and hypothermia induce divergent changes in mean CBF and CGU. However, local analysis demonstrates a well-maintained linear relationship between LCBF and LCGU during normothermic and hypothermic anesthesia. PMID- 10835461 TI - Brain tissue sodium is a ticking clock telling time after arterial occlusion in rat focal cerebral ischemia. AB - BACKGROUND AND PURPOSE: Many patients with acute stroke are excluded from receiving thrombolysis agents within the necessary time limit (3 or 6 hours from stroke onset) because they or their family members are unable provide the time of stroke onset. Brain tissue sodium concentration ([Na(+)]) increases gradually and incessantly during the initial hours of experimental focal cerebral ischemia but only in severely damaged brain regions. We propose that this steady increase in [Na(+)] can be used to estimate the time after arterial occlusion in the rat middle cerebral artery occlusion model of ischemic stroke. METHODS: Sixteen anesthetized Sprague-Dawley rats underwent permanent middle cerebral artery occlusion combined with bilateral common artery occlusion. After 100 to 450 minutes, diffusion-weighted MRI was used to generate apparent diffusion coefficient (ADC) maps, cerebral blood flow (CBF) was determined with (14)C iodoantipyrine (in a subset of 7 animals), and the brain was frozen. Autoradiographic CBF sections and punch samples for Na(+) analysis were obtained from the brain at the same level of the MR image. Severely at risk regions were identified with an ADC of <520 microm(2)/s and, in the subset, with both ADC of <520 microm(2)/s and CBF of <40 mL. 100 g(-1). min(-1). RESULTS: Both CBF and the ADC dropped quickly and remained stable in the initial hours after ischemic onset. Linear regression revealed strong linearity between [Na(+)] and time after onset, with a slope of 0.95 or 1.00 (mEq/kg DW)/min, with both ADC and ADC-plus CBF criteria, respectively. The 95% CIs at 180 and 360 minutes were between 41 and 52 minutes. CONCLUSIONS: The time after ischemic onset can be estimated with this 2-step process. First, ADC and CBF are used to identify severely endangered regions. Second, the [Na(+)] in these regions is used to estimate time after onset. The favorable 95% CIs at the time limits for thrombolytic therapy and the availability of measurements of ADC, CBF, and [Na(+)] in humans through the use of MRI suggest that this time-estimation scheme could be used to assess the appropriateness of thrombolysis for patients who do not know when the stroke occurred. PMID- 10835463 TI - Integrin alpha(IIb)beta(3) inhibitor preserves microvascular patency in experimental acute focal cerebral ischemia. AB - BACKGROUND AND PURPOSE: Platelets become activated and accumulate in brain microvessels of the ischemic microvascular bed after experimental focal cerebral ischemia. The binding of glycoprotein IIb/IIIa (integrin alpha(IIb)beta(3)) on platelets to fibrinogen is the terminal step in platelet adhesion and aggregation. This study tests the hypothesis that inhibition of platelet fibrin(ogen) interactions may prevent microvascular occlusion after experimental middle cerebral artery occlusion (MCA:O). METHODS: TP9201 is a novel Arg-Gly-Asp (RGD)-containing integrin alpha(IIb)beta(3) inhibitor. Microvascular patency after 3-hour MCA:O and 1-hour reperfusion within the ischemic and nonischemic basal ganglia was compared in adolescent male baboons who received high-dose TP9201 (group A: IC(80) in heparin, n=4), low-dose TP9201 (group B: IC(30) in heparin, n=4), or no treatment (group C: n=4) before MCA:O. RESULTS: After MCA:O, microvascular patency decreased significantly in group C. However, in the ischemic zones of groups A and B compared with group C, patencies were significantly greater in the 4.0- to 7. 5-microm-diameter (capillary) and 7.5- to 30.0-microm-diameter vessels (2P<0.05). A dose-dependent increase in hemorrhagic transformation was seen in group A (3 of 4 animals) compared with group B (1 of 4 animals), and no hemorrhage was visible in group C (chi(2) analysis for trend, P<0.05). CONCLUSIONS: Platelet activation contributes significantly to ischemic microvascular occlusion. Occlusion formation may be prevented by this RGD integrin alpha(IIb)beta(3) inhibitor at a dose that does not produce clinically significant parenchymal hemorrhage. The effect of microvascular patency on neuron recovery can now be tested. PMID- 10835464 TI - Delayed systemic administration of PACAP38 is neuroprotective in transient middle cerebral artery occlusion in the rat. AB - BACKGROUND AND PURPOSE: Many substances have been shown to reduce brain damage in models of stroke, but mainly when given either before or shortly after the onset of ischemia. Delayed systemic administration of pituitary adenylate cyclase activating polypeptide (PACAP) has been shown to attenuate the neuronal damage in the hippocampus in a model of global ischemia in rats. The present study examined the neuroprotective action of delayed systemic administration of PACAP38 in a model of transient focal ischemia produced by middle cerebral artery occlusion (MCAO) in rats. METHODS: We administered PACAP38 as an intravenous bolus (20 nmol/kg body wt) followed by an intravenous infusion for 48 hours using a micro osmotic pump at a rate of 160 pmol/microL per hour, beginning 4, 8, or 12 hours after a 2-hour transient MCAO using a filament model. The size of the infarct was determined by examining 2-mm-thick brain sections stained with triphenyltetrazolium chloride, followed by image analysis. Control animals received intravenously 0.1% bovine serum albumin in 0.9% saline as a bolus and infusion at the same time intervals. RESULTS: The administration of PACAP38 beginning 4 hours after MCAO significantly reduced the infarct size by 50.88%. Treatment with PACAP38 starting 8 or 12 hours after the onset of ischemia did not result in a significant reduction of the infarct size, although infarct volumes tended to be smaller than in the control groups. CONCLUSIONS: Systemic administration of PACAP38 should be clinically useful for reducing brain damage resulting from stroke even when administration is delayed for several hours. PMID- 10835465 TI - Amnesia due to fornix infarction. AB - Background and Purpose-The fornix connects various structures involved in memory. We report a patient with anterograde amnesia after an acute ischemic infarct in the anterior fornix. Case Description-A 71-year-old female with acute-onset amnesia had neuroimaging studies showing ischemic infarction of both columns and the body of the fornix and the genu of the corpus callosum. Neuropsychological evaluation revealed anterograde amnesia without evidence of callosal disconnection. The patient showed marked improvement in her memory function on the follow-up visit. Conclusions-Amnesia in this case is likely due to infarction of the anterior fornix structures. PMID- 10835466 TI - Endovascular treatment of dural sinus thrombosis with rheolytic thrombectomy and intra-arterial thrombolysis. AB - BACKGROUND AND PURPOSE: Cerebral venous thrombosis is a rare entity that can be difficult to manage. Intrasinus thrombolysis is an increasingly applied intervention, but this modality carries an increased risk of hemorrhage. We describe for the first time an option with a potentially lower incidence of intracranial bleeding, the combination of the AngioJet rheolytic thrombectomy catheter with intra-arterial thrombolysis, in 2 patients with extensive dural sinus thromboses, preexisting intracranial hemorrhage, and severe progressive neurological deficits despite heparin therapy. METHODS: Four procedures were performed in 2 patients with thromboses in the superior sagittal and transverse sinuses (right in 1 patient and bilateral in 1 patient) and cortical veins. Rheolytic thrombectomy was performed in the sigmoid, transverse, straight, and superior sagittal sinuses; this technique involves the use of the Bernoulli effect to create a vacuum that fragments and aspirates thrombus. For associated persistent cortical vein thromboses, low-dose intra-arterial thrombolysis was used. RESULTS: Both patients had excellent angiographic results with sinus reopening after rheolytic thrombectomy and cortical vein reopening after intra arterial thrombolysis. Follow-up CT showed no change in 1 patient and increased preexisting intracranial hemorrhage in the other. One patient had a negative hypercoagulable workup, and the other patient had probable anti-phospholipid antibody syndrome. At 6 months, both patients had excellent clinical outcome with no neurological deficits except mild short-term memory loss in 1 patient. CONCLUSIONS: The combination of rheolytic thrombectomy with intra-arterial thrombolysis is a treatment modality that allows accelerated recanalization of occluded dural sinuses and cerebral veins with lower doses of thrombolytic agents. PMID- 10835467 TI - Stress-related primary intracerebral hemorrhage: autopsy clues to underlying mechanism. AB - BACKGROUND: Research into the causes of small-vessel stroke has been hindered by technical constraints. Cases of intracerebral hemorrhage occurring in unusual clinical contexts suggest a causal role for sudden increases in blood pressure and/or cerebral blood flow. CASE DESCRIPTION: We describe a fatal primary thalamic/brain stem hemorrhage occurring in the context of sudden emotional upset. At autopsy, the brain harbored several perforating artery fibrinoid lesions adjacent to and remote from the hematoma as well as old lacunar infarcts and healed destructive small-vessel lesions. CONCLUSIONS: We postulate that the emotional upset caused a sudden rise in blood pressure/cerebral blood flow, mediating small-vessel fibrinoid necrosis and rupture. This or a related mechanism may underlie many small-vessel strokes. PMID- 10835468 TI - Outcome measures in acute stroke trials: a systematic review and some recommendations to improve practice. AB - BACKGROUND: There is little consistency in the measurement of outcome in acute stroke trials, and this may complicate interpretation of the results and reduce the likelihood of detecting worthwhile drug effects. This study aims to investigate empirically the measures used to date and to give recommendations for future studies. SUMMARY OF COMMENT: A systematic review of all published randomized studies of acute stroke drug intervention was undertaken, and the measures used were recorded. Fifty-one studies involving 57 214 subjects were identified. These studies used 14 different measures of impairment, 11 different measures of activity, 1 measure of "quality of life," and 8 miscellaneous other measures. Timing of outcome assessments varied from 1 week to 1 year, with the modal time being 3 months. Many studies used ordinal measures but dichotomized results for analysis. Of the 51 studies included in the review, only 21 demonstrated benefit with the defined primary outcome measure. In several studies, however, post hoc analysis using varied outcome measures or varied cut points for dichotomizing outcomes resulted in positive results, whereas the primary study analysis failed to do so. CONCLUSIONS: There is no consensus on the level of outcome to be used, the method of measurement to be used, or the most appropriate timing of the assessment. It is recommended that future studies should include extended/instrumental activities and advanced mobility as components of the primary outcome measure, with outcome assessment being undertaken at 6 months. New initiatives in developing stroke-specific outcomes may address some of the current problems in the assessment of stroke outcomes PMID- 10835469 TI - Systematic comparison of the early outcome of angioplasty and endarterectomy for symptomatic carotid artery disease. AB - BACKGROUND AND PURPOSE: Endoluminal treatment is being increasingly used for carotid artery disease. The aim of this study was to compare the stroke and death risk within 30 days of endovascular treatment or endarterectomy for symptomatic carotid artery disease. METHODS: systematic comparison of the 30-day outcome of angioplasty with or without stenting and endarterectomy for symptomatic carotid artery disease reported in single-center studies, published since 1990, was performed. RESULTS: Thirty-three studies (13 angioplasty and 20 carotid endarterectomy) were included in this analysis. Carotid stents were deployed in 44% of angioplasty patients. Mortality within 30 days of angioplasty was 0.8% compared with 1.2% after endarterectomy (OR 0.68, 95% CI 0.43 to 1.05; P=0.6). The stroke rate was 7.1% for angioplasty and 3.3% for endarterectomy (OR 2.22, CI 1.62 to 3.04; P<0.001), while the risk of fatal or disabling stroke was 3.2% and 1.6%, respectively (OR 2.09, CI 1.3 to 3.33; P<0.01). The risk of stroke or death was 7.8% for angioplasty and 4% for endarterectomy (OR 2.02, CI 1.49 to 2.75; P<0.001), while disabling stroke or death was 3.9% after angioplasty and 2.2% after endarterectomy (OR 1.86, CI 1.22 to 2.84; P<0.01). CONCLUSIONS: In the treatment of symptomatic carotid artery disease, the risk of stroke is significantly greater with angioplasty than carotid endarterectomy. At present, carotid angioplasty is not recommended for the majority of patients with symptomatic carotid artery disease. PMID- 10835470 TI - Critical appraisal of the design and reporting of studies of imaging and measurement of carotid stenosis. AB - BACKGROUND AND PURPOSE: Several hundred studies have been published over the last few years on imaging and measurement of carotid stenosis. Despite all this research, there is still no consensus about how best to image and measure stenosis. One possible explanation for this is that many of the studies have not been large enough or methodologically sound enough to allow useful conclusions to be drawn. We aimed to assess the design and methods of a random sample of published studies of imaging and measurement of carotid stenosis using 9 simple criteria. METHODS: A formal literature search was performed for studies of imaging and measurement of carotid stenosis. Two subsets were randomly selected for detailed assessment: 20 studies published before 1991 and 20 published between 1993 and 1997 (some years after the initial publication of the ECST and NASCET trials). The criteria used to assess the selected studies were as follows: prospective rather than retrospective study design; patient selection based on a consecutive series or a random sample; adequate detail of study population; adequate detail of imaging techniques; inclusion of all investigations, ie, patients with poor-quality imaging were not excluded; blinded assessment of images; adequate detail of derivation of measurement of stenosis from images or data; adequate data on the reproducibility of measurements of stenosis; and study powered according to a sample-size calculation. RESULTS: There were many basic methodological deficiencies in both subsets of studies, with relatively little evidence of improvement with time. For example, only 33% of studies were prospective, only 45% studied a consecutive or random selection of patients, and only 38% reported any data on the reproducibility of measurements. More than half of the studies satisfied < or =4 of the 9 quality criteria. However, there was considerable variation between studies, with 7 studies satisfying > or = 7 criteria and 10 studies satisfying < or =2. No study was based on a sample-size calculation. The number of patients studied was often small, particularly in the more recent studies: median sample size was 100 in the 1970-1990 studies and 58 in the 1993-1997 studies (P<0.0001). CONCLUSIONS: The design and reporting of published studies of imaging and measurement of carotid stenosis are poor and have not improved much in recent years. The majority of published studies are not of a sufficient standard to enable the results to be used to inform clinical practice. The utility of future studies could be improved considerably by better adherence to 9 simple methodological guidelines. PMID- 10835471 TI - CVA: reducing the risk of a confused vascular analysis. The Feinberg lecture. PMID- 10835472 TI - Diffusion-weighted magnetic resonance imaging in brain death. PMID- 10835473 TI - Ultrasonic assessment of brain perfusion. PMID- 10835474 TI - Hyperglycemia and extracellular glutamate in the ischemic brain. PMID- 10835475 TI - Is transcranial magnetic stimulation of the motor cortex a prognostic tool for motor recovery after stroke? PMID- 10835476 TI - Pharmacological preconditioning with lipopolysaccharide in the brain. PMID- 10835477 TI - tPA: a rural network experience. PMID- 10835479 TI - Major ongoing stroke trials. PMID- 10835478 TI - Abstracts of literature PMID- 10835480 TI - Anopheles stephensi Dox-A2 shares common ancestry with genes from distant groups of eukaryotes encoding a 26S proteasome subunit and is in a conserved gene cluster. AB - The sequence of a cloned Anopheles stephensi gene showed 72% inferred amino acid identity with Drosophila melanogaster Dox-A2 and 93% with its putative ortholog in Anopheles gambiae. Dox-A2 is the reported but herein disputed structural locus for diphenol oxidase A2. Database searches identified Dox-A2 related gene sequences from 15 non-insect species from diverse groups. Phylogenetic trees based on alignments of inferred protein sequences, DNA, and protein motif searches and protein secondary structure predictions produced results consistent with expectations for genes that are orthologous. The only inconsistency was that the C-terminus appears to be more primitive in the yeasts than in plants. In mammals, plants, and yeast these genes have been shown to code for a non-ATPase subunit of the PA700 (19S) regulatory complex of 26S proteasome. The analyses indicated that the insect genes contain no divergent structural features, which taken within an appraisal of all available data, makes the reported alternative function highly improbable. A plausible additional role, in which the 26S proteasome is implicated in regulation of phenol oxidase, would also apply to at least the mammalian genes. No function has yet been reported for the other included sequences. These were from genome projects and included Caenorhabiditus elegans, Arabidopsis thaliana, Fugu rubripes, and Toxoplasma gondii. A consensus of the results predicts a protein containing exceptionally long stretches of helix with a hydrophilic C-terminus. Phosphorylation site motifs were identified at two conserved positions. Possible SRY and GATA-1 binding motifs were found at conserved positions upstream of the mosquito genes. The location of A. stephensi Dox-A2 was determined by in situ hybridization at 34D on chromosome arm 3R. It is in a conserved gene cluster with respect to the other insects. However, the A. stephensi cluster contains a gene showing significant sequence identity to human and pigeon carnitine acetyltransferase genes, therefore showing divergence with the distal end of the D. melanogaster cluster. PMID- 10835481 TI - Using alu J elements as molecular clocks to trace the evolutionary relationships between duplicated HLA class I genomic segments. AB - The class I region of the major histocompatibility complex contains two subgenomic blocks (250-350 kb each), known as the alpha and beta blocks. These blocks contain members of multicopy gene families including HLA class I, HERV-16 (previously called P5 sequences), and PERB11 (MIC). We have previously shown that each block consists of imperfect duplicated segments (duplicons) containing linked members of different gene families, retroelements and transposons that have coevolved as part of two separate evolutionary events. Another region provisionally designated here as the kappa block is located between the alpha and the beta blocks and contains HLA-E, -30, and -92, HERV-16 (P5.3), and PERB11.3 (MICC) within about 250 kb of sequence. Using Alu elements to trace the evolutionary relationships between different class I duplicons, we have found that (a) the kappa block contains paralogous (duplicated) Alu J sequences and other retroelement patterns more in common with the beta than the alpha block; (b) the retroelement pattern associated with the HLA-E duplicon is different from all other HLA class I duplicons, indicating a more complex evolution; (c) the HLA 92 duplicon, although substantially shorter, is closely related in sequence to the HLA-B and -C duplicons; (d) two of the six paralogous Alu J elements within the HLA-B and -C duplicons are associated with the HLA-X duplicon, confirming their evolutionary relationships within the beta block; and (e) the paralogous Alu J elements within the alpha block are distinctly different from those identified within the beta and kappa blocks. The sequence conservation and location of duplicated (paralogous) Alu J elements in the MHC class I region show that the beta and kappa blocks have evolved separately from the alpha block beginning at a time before or during the evolution of Alu J elements in primates. PMID- 10835482 TI - Molecular evolution of apoptotic pathways: cloning of key domains from sponges (Bcl-2 homology domains and death domains) and their phylogenetic relationships. AB - Cells from metazoan organisms are eliminated in a variety of physiological and pathophysiological processes by apoptosis. In this report, we describe the cloning and characterization of molecules from the marine sponges Geodia cydonium and Suberites domuncula, whose domains show a high similarity to those that are found in molecules of the vertebrate Bcl-2 superfamily and of the death receptors. The Bcl-2 proteins contain up to four Bcl-2 homology regions (BH). Two Bcl-2-related molecules have been identified from sponges that are provided with two of those regions, BH1 and BH2, and are termed Bcl-2 homology proteins (BHP). The G. cydonium molecule, BHP1_GC, has a putative size of 28,164, while the related sequence from S. domuncula, BHP1_SD, has a M(r) of 24,187. Phylogenetic analyses of the entire two sponge BHPs revealed a high similarity to members of the mammalian Bcl-2 superfamilies and to the Caenorhabditis elegans Ced-9. When the two domains, BH1 and BH2, are analyzed separately, again the highest similarity was found to the members of the Bcl-2 superfamily, but a clearly lower relationship to the C. elegans BH1 and BH2 domains in Ced-9. In unrooted phylogenetic trees the sponge BH1 and BH2 are grouped among the mammalian sequences and are only distantly related to the C. elegans BH domains. The analysis of the gene structure of the G. cydonium BHP showed that the single intron present is located within the BH2 domain at the same position as in C. elegans and rat Bcl-x(L). In addition, a sponge molecule comprising two death domains has been characterized from G. cydonium. The two death domains of the potential proapoptotic molecule GC_DD2, M(r) 24,970, share a high similarity with the Fas-FADD/MORT1 domains. A death domain-containing molecule has not been identified in the C. elegans genome. The phylogenetic analysis revealed that the sponge domain originated from an ankyrin building block from which the mammalian Fas-FADD/MORT1 evolved. It is suggested that the apoptotic pathways that involve members of the Bcl-2 superfamily and of the death receptors are already present in the lowest metazoan phylum, the Porifera. PMID- 10835483 TI - Phylogenetic relationships of Acanthocephala based on analysis of 18S ribosomal RNA gene sequences. AB - Acanthocephala (thorny-headed worms) is a phylum of endoparasites of vertebrates and arthropods, included among the most phylogenetically basal tripoblastic pseudocoelomates. The phylum is divided into three classes: Archiacanthocephala, Palaeacanthocephala, and Eoacanthocephala. These classes are distinguished by morphological characters such as location of lacunar canals, persistence of ligament sacs in females, number and type of cement glands in males, number and size of proboscis hooks, host taxonomy, and ecology. To understand better the phylogenetic relationships within Acanthocephala, and between Acanthocephala and Rotifera, we sequenced the nearly complete 18S rRNA genes of nine species from the three classes of Acanthocephala and four species of Rotifera from the classes Bdelloidea and Monogononta. Phylogenetic relationships were inferred by maximum likelihood analyses of these new sequences and others previously determined. The analyses showed that Acanthocephala is the sister group to a clade including Eoacanthocephala and Palaeacanthocephala. Archiacanthocephala exhibited a slower rate of evolution at the nucleotide level, as evidenced by shorter branch lengths for the group. We found statistically significant support for the monophyly of Rotifera, represented in our analysis by species from the clade Eurotatoria, which includes the classes Bdelloidea and Monogononta. Eurotatoria also appears as the sister group to Acanthocephala. PMID- 10835484 TI - Formation of the Japanese Carabina fauna inferred from a phylogenetic tree of mitochondrial ND5 gene sequences (Coleoptera, carabidae). AB - Phylogenetic analyses based on the mitochondrial ND5 gene comparisons and the geohistory of the Japanese Islands suggest that each Japanese species belonging to the subtribe Carabina has its own history for the establishment of its present habitat in the Japanese Islands. It can be roughly classified into two categories: (1) species which were derived from the ancestry that inhabited ancient Japan at the time of its split from the Eurasian Continent [ca. 15 million years ago (MYA)], followed by diversification within the Japanese Islands; and (2) species which invaded Hokkaido from the Eurasian Continent through land-bridges from Sakhalin and/or the Kuriles or from western Japan from the Korean Peninsula during the glacial era (<2 MYA). PMID- 10835485 TI - Molecular characterization of STAT5A- and STAT5B-encoding genes reveals extended intragenic sequence homogeneity in cattle and mouse and different degrees of divergent evolution of various domains. AB - The STAT transcription factors form a family of signal transducers and activators of transcription. We sequenced the bovine STAT5B cDNA and both STAT5-encoding genes, STAT5A and STAT5B, representing the first complete description of any STAT5-encoding gene. DNA fiber FISH hybridization revealed that the genes reside only 40 kbp apart on BTA19. Both genes are segmented into 19 exons and all but two of the homologous exons are of equal size. The genes harbor a central block of nearly identical DNA sequence (97.5% sequence identity over 3373 bp), spanning from intron 5 to intron 9. Isolation and sequencing of the homologous segments from mouse revealed the same unusually high degree of intronic sequence conservation in these segments of the murine STAT5-encoding genes. However, the respective sequences are completely divergent between the two species. A comparison of the inter- and intragenic cDNA sequence preservation at nonsynonymous sites reveals that the DNA-binding domain is under the strongest selection pressure for both intergenic and factor-specific intragenic sequence preservation. The so-called "SH3" segment of the linker domain, in contrast, shows species-specific sequence identity in all but one amino acid residues in both factors, in cattle, human, and mouse. This indicates that the same species specific selection pressure occurs on the linker domain from both factors, STAT5A and STAT5B. Thus, the comparison of evolutionary selection pressures resting on various domains suggests that the DNA-binding domain might contribute to differential DNA binding of STAT5A and STAT5B factors, while both might interact equally well with other cellular factors through a segment of the linker domain. PMID- 10835486 TI - Evolutionary rate acceleration of cytochrome c oxidase subunit I in simian primates. AB - We present an analysis of the evolutionary rates of the cytochrome c oxidase subunit I genes of primates and other mammals. Five primate genes were sequenced, and this information was combined with published data from other species. The sequences from simian primates show approximately twofold increases in their nonsynonymous substitution rate compared to those from other primates and other mammals. The species range and the overall magnitude of this rate increase are similar to those previously identified for the cytochrome c oxidase subunit II and cytochrome b genes. PMID- 10835487 TI - The mitochondrial genome of the sperm whale and a new molecular reference for estimating eutherian divergence dates. AB - Extant cetaceans are systematically divided into two suborders: Mysticeti (baleen whales) and Odontoceti (toothed whales). In this study, we have sequenced the complete mitochondrial (mt) genome of an odontocete, the sperm whale (Physeter macrocephalus), and included it in phylogenetic analyses together with the previously sequenced complete mtDNAs of two mysticetes (the fin and blue whales) and a number of other mammals, including five artiodactyls (the hippopotamus, cow, sheep, alpaca, and pig). The most strongly supported cetartiodactyl relationship was: outgroup,((pig, alpaca), ((cow, sheep),(hippopotamus,(sperm whale,(baleen whales))))). As in previous analyses of complete mtDNAs, the sister group relationship between the hippopotamus and the whales received strong support, making both Artiodactyla and Suiformes (pigs, peccaries, and hippopotamuses) paraphyletic. In addition, the analyses identified a sister-group relationship between Suina (the pig) and Tylopoda (the alpaca), although this relationship was not strongly supported. The paleontological records of both mysticetes and odontocetes extend into the Oligocene, suggesting that the mysticete and odontocete lineages diverged 32-34 million years before present (MYBP). Use of this divergence date and the complete mtDNAs of the sperm whale and the two baleen whales allowed the establishment of a new molecular reference, O/M-33, for dating other eutherian divergences. There was a general consistency between O/M-33 and the two previously established eutherian references, A/C-60 and E/R-50. Cetacean (whale) origin, i.e., the divergence between the hippopotamus and the cetaceans, was dated to approximately 55 MYBP, while basal artiodactyl divergences were dated to >/=65 MYBP. Molecular estimates of Tertiary eutherian divergences were consistent with the fossil record. PMID- 10835488 TI - Isolation of two novel genes, down-regulated in gastric cancer. AB - Using a differential display technique, we identified two genes that are down regulated in human gastric cancer tissue as compared to normal gastric mucosa. The down-regulated expression of these genes in gastric cancer tissue was confirmed by northern blotting analysis and RT-PCR. One, CA11, was a novel gene expressed predominantly in the stomach and was depleted in all of the gastric cancer cell lines examined. The other gene, GC36, was homologous to the digestive tract-specific calpain gene, nCL-4. The expression of both GC36 and nCL-4 was suppressed or depleted in gastric cancer cell lines of differentiated and poorly differentiated types. This is the first report of genes, the expression of which is down-regulated with considerable frequency in gastric cancer. PMID- 10835490 TI - Long-term effect of Helicobacter pylori infection on serum pepsinogens. AB - Serum pepsinogen values are markers of gastric mucosal status and of gastric cancer risk. The effect of Helicobacter pylori infection and sibship size on change of serum pepsinogen values over a seven-year span was investigated. Data from 2584 subjects with phlebotomy were analyzed both in 1989 and in 1996. The subjects were classified by H. pylori serology and sibship size (1 - 3 vs. 4 and more). Pepsinogen I (PG I) to II (PG II) ratio in '96 minus that in '89 was defined as DeltaPG I / II and compared among the groups. DeltaPG I / II was lower and decrease of PG I / II was more frequent among H. pylori-positive subjects than among negative subjects. The difference was owing to a decrease of PG I in all subjects and owing to an increase of PG II in those not younger than 30 years in '89. In H. pylori-positive subjects, those with a larger sibship size showed lower DeltaPG I / II and higher frequency of PG I / II decline. H. pylori infection exerts a reducing effect on PG I / II during the seven-year span. The effect of H. pylori is stronger among those with a larger sibship size, who are expected to have been infected with H. pylori in childhood. Inducing atrophy of gastric mucosa, which is reflected by a decline of PG I / II, may be one of the mechanisms through which H. pylori elevates the risk of gastric cancer. PMID- 10835489 TI - Effects of three-month oral supplementation of beta-carotene and vitamin C on serum concentrations of carotenoids and vitamins in middle-aged subjects: a pilot study for a randomized controlled trial to prevent gastric cancer in high-risk Japanese population. AB - Prior to a randomized controlled trial to prevent gastric cancer by oral supplementation of beta-carotene and vitamin C in a high-risk Japanese population, we examined the serum response to three-month oral supplementation of beta-carotene (0, 3, 30 mg / day) and vitamin C (0, 50, 1000 mg / day) by a three by-three factorial design using 54 subjects (age range = 40 - 69 years). Serum concentrations of carotenoids, alpha-tocopherol, and ascorbic acid were examined at baseline, and one, two, and three-month points. Both serum beta-carotene and ascorbic acid were significantly higher in high-dose groups than in each placebo group during the supplementation. The serum beta-carotene increased gradually (597 - 830% increase) during the study, whereas the serum ascorbic acid reached nearly a steady-state at the one-month point and remained stable thereafter (88 - 95% increase). No statistically significant interaction between beta-carotene and vitamin C supplementations was observed either for serum beta-carotene or for serum ascorbic acid. Among carotenoids and alpha-tocopherol examined, serum lycopene in the high-dose beta-carotene group was significantly higher than in the placebo group at all points. No unfavorable change in carotenoids and alpha tocopherol was observed in any group. PMID- 10835491 TI - Inhibitory effects of 1'-acetoxychavicol acetate on N-Nitrosobis(2-oxopropyl) amine-induced initiation of cholangiocarcinogenesis in Syrian hamsters. AB - The influence of 1'-acetoxychavicol acetate (ACA) during the initiation stage was investigated in the N-nitrosobis(2-oxopropyl)amine (BOP)-initiated hamster tumorigenesis model. Ninety male 5-week-old hamsters were divided into three groups, each consisting of 30 animals, and s.c. injected with 20 mg / kg of BOP twice with a one-week interval. Groups 1 through 3 were fed diet supplemented with ACA at concentrations of 500, 100 and 0 ppm, respectively, for 3 weeks starting one week before the first carcinogen application. At the termination of experimental week 54, the total incidence and multiplicity of cholangiocellular adenomas and carcinomas in group 1 (17.9% and 0.3 < 0.9) were significantly (P < 0.05 and P < 0.01) decreased as compared to the group 3 values (50.0% and 0.7 < 0.8). The ACA treatments also showed a tendency to reduce the development of preneoplastic lesions in the pancreas, a main target organ of BOP, although this was not statistically significant. Our results thus indicate that ACA exerts an inhibitory effect on BOP-induced cholangiocarcinogenesis in hamsters. Taken together with previous findings of inhibited colon, oral and skin carcinogenesis in rats and mice, they suggest that ACA is a candidate chemopreventive agent with a wide spectrum of activity. PMID- 10835492 TI - Different mutation frequencies and spectra among organs by N-methyl-N-nitrosourea in rpsL (strA) transgenic mice. AB - The frequencies and spectra of N-methyl-N-nitrosourea (MNU)-induced in vivo somatic mutations were determined in rpsL (strA) transgenic mice. The wild-type rpsL gene, which exhibits a streptomycin-sensitive (Sm(S)) phenotype, was used as the rescue marker gene. Studies of mutation spectra among different organs and tissues were simplified using this system because of the short coding sequence (375 bp) of the rpsL gene. MNU administration to transgenic mice significantly elevated the mutation frequencies in various adult organs. Two distinctive patterns of mutation spectrum were observed, depending on the organs tested. Mutations derived from labile organs (spleen and thymus) were predominantly G:C to A:T transitions, as expected for MNU mutagenesis. Stable organs like the liver and brain, however, carried many fewer G:C to A:T transitions but significantly more single base deletions, of which the spectrum was very similar to that of background mutations in the rpsL transgenic mice. This spectrum difference among more and less proliferating organs was confirmed by the predominant occurrence of G:C to A:T transitions in fetal liver cells exposed to transplacental MNU treatment. In addition, most (approximately 90%) of the G:C to A:T transitions induced by MNU were detected in the first nucleotide of some 5'-G-(C or G)-3' sequences, many of which corresponded to the middle guanine residue of 5'-purine G-(C or G)-3' sequences. It is thus suggested that at particular sites, the neighboring bases in both the 5' side and 3' side seem to influence either the susceptibility to DNA damage or the ability to repair MNU-induced lesions. PMID- 10835493 TI - MDM2 overexpression with alteration of the p53 protein and gene status in oral carcinogenesis. AB - In this study, to better understand the mechanism of oral squamous cell carcinoma (SCC) carcinogenesis, alterations of the p53 gene and overexpression of MDM2 and p53 were analyzed in 38 oral SCC samples. Twelve of the 38 specimens revealed mutant-type p53. Moreover, coexpression of MDM2 and p53 was found most frequently in dysplastic lesions (P < 0.05). Expression of MDM2 and p53 was significantly increased in accordance with the histological progression of multistep carcinogenesis (P < 0.05). No significant correlation was found between the expression of MDM2 and the alteration of p53 protein or p53 gene status. MDM2 overexpression with mutant p53 was significantly associated with poorly differentiated SCCs (P < 0.05) and tumor stages III and IV of oral SCCs (P < 0.05). These results suggest that MDM2 overexpression is an early event in oral carcinogenesis through the functional inactivation of the wild-type p53, and corresponding alterations of MDM2 and p53 contribute to the oral carcinogenesis. We propose that it would be clinically more instructive to evaluate MDM2 overexpression combined with p53 gene status, compared to the evaluation of either MDM2 or p53 alteration alone. PMID- 10835494 TI - Allelic imbalances on chromosome 20 in human transitional cell carcinoma. AB - One determinant of the survival time of cancer-bearing patients may be genetic factors. In chemically induced bladder cancers of mice, differences in survival time have been observed among several inbred strains. Genetic analyses of such differences in crosses between C57BL / 6 and NON mice revealed that the survival period is determined by two quantitative trait loci on mouse chromosomes 6 and 2, respectively. We explored the possibility that genetic alterations may be observed in the syntenic conserved chromosomal regions of human transitional cell carcinoma corresponding to mouse chromosomes 6 and 2. Human chromosome 7, containing a region syntenic to mouse chromosome 6, is reported to harbor frequent genetic alterations in bladder cancers. In this study, we investigated 70 human urothelial cancers for possible genetic alterations on human chromosome 20p and 20q containing regions syntenic to mouse chromosome 2. Allelic imbalances were observed in 22 cases (31.4%) on 20p and 18 cases (25.7%) on 20q. Those allelic imbalances, however, did not show a direct correlation with the prognosis of the patients. Higher grade tumors tended to show more frequent imbalances on chromosome 20; however, this tendency was not significant. PMID- 10835495 TI - Infrequent mutation of the hBUB1 and hBUBR1 genes in human lung cancer. AB - Mitotic checkpoint defects of the cell cycle have been implicated in the development of human cancers. Since hBUB1 and hBUBR1, whose products function in the spindle checkpoint pathway, have been shown to be mutated in a subset of colon cancers with chromosomal instability, we investigated the contribution of these genes to lung cancer development. One hundred and two lung cancer (50 small cell lung cancers and 52 non-small cell lung cancers) and 4 mesothelioma cell line DNAs were analyzed by Southern blot analysis, but no rearrangements or deletions of hBUB1 and hBUBR1 were detected. Using single strand conformation polymorphism analysis, we studied all the 25 exons except exon 1 of the hBUB1 gene in 88 lung cancer DNAs. One lung cancer cell line, NCI-H345, showed a single nucleotide substitution, which resulted in an Arg-to-Gln change at codon 209 (CGA to CAA). Eleven cell line DNAs exhibited a single nucleotide polymorphism in intron 9 of hBUB1, all of which were heterozygous. Similar mutation analysis of hBUBR1 in 47 lung cancer cell line cDNAs revealed a frequent polymorphism at codon 349 (CAA to CGA) leading to a substitution of Gln to Arg but no mutations. Northern blot analyses showed that both hBUB1 and hBUBR1 genes were expressed in all of 31 lung cancer cell lines tested with no significant difference in the expression level. Our results suggest that alterations in hBUB1 and hBUBR1 rarely contributed to the genetic change of lung cancers. PMID- 10835496 TI - Changes in expression of estrogen receptors alpha and beta in relation to progesterone receptor and pS2 status in normal and malignant endometrium. AB - To clarify changes in estrogen receptor (ER) alpha and ERbeta during endometrial tumorigenesis, 48 endometrial carcinomas (endometrioid type), as well as 40 samples of normal endometrial tissue, were investigated using a combination of reverse-transcription and polymerase chain reaction with Southern blot hybridization and western blot assays, and the results were compared with findings for progesterone receptor (PR) and pS2 mRNA status. In addition, 166 carcinomas were also examined for immunohistochemistry, along with 171 normal specimens. Relative amounts of ERalpha at both mRNA and protein levels were significantly greater than those for ERbeta in normal and malignant endometrial lesions. ERalpha mRNA showed a stepwise decrease from normal or grade (G) 1 through to G3 tumor lesions, in line with changes in the protein levels, in contrast to ERbeta mRNA or protein expression, which did not alter, suggesting a shift in the ratio of the two ER subtypes during endometrial tumorigenesis. PR mRNA expression was significantly correlated with ERalpha, but not ERbeta mRNA status. Although significantly higher expression of pS2 mRNA or protein was observed in carcinomas than in the normal cases, there was no apparent association with the ER status. The findings suggest that alteration in estrogen signaling pathways may occur during endometrial tumorigenesis, and provide evidence that ERalpha expression may play an important role in the regulation of PR, but not pS2 expression in normal and malignant endometrium. PMID- 10835497 TI - BRCA1 expression status in relation to DNA methylation of the BRCA1 promoter region in sporadic breast cancers. AB - To understand the biological role of BRCA1 in sporadic breast cancers, the relationship between DNA methylation of the BRCA1 promoter region and BRCA1 expression was studied using molecular biological and immunohistochemical methods. Furthermore, BRCA1 expression was compared with the expression of various cell cycle regulatory proteins and the morphological nuclear grade of cancer cells. Of 32 sporadic breast cancers investigated in this study, 10 (31%) revealed DNA methylation of the BRCA1 promoter region. The expression of BRCA1 was observed in the nuclei of cancer cells and 18 (56%) of 32 cancers were positive for BRCA1 immunoreactivity. Breast cancers with BRCA1 methylation lacked BRCA1 expression, except for only three cancers, and there was a significant inverse relationship between BRCA1 methylation and its expression in sporadic breast cancers (P = 0.043). Compared with the expression of various cell cycle regulatory proteins, breast cancers with BRCA1 methylation showed decreased expression of estrogen receptor (P = 0. 016) and p27 (P = 0.018) and increased expression of p21 (P = 0.011). Furthermore, breast cancers without BRCA1 expression or with BRCA1 methylation had a tendency to contain nuclei with higher grade. These findings indicate that BRCA1 methylation might greatly influence its expression and BRCA1 expression might play an important role in cell cycle regulation and influence the grade of malignancy of sporadic breast cancers. PMID- 10835498 TI - Ras and signal transducer and activator of transcription (STAT) are essential and sufficient downstream components of Janus kinases in cell proliferation. AB - Cytokines exert their activities in cell growth and differentiation by binding specific cell membrane receptors. Janus kinases (JAKs) are cytoplasmic protein tyrosine kinases that physically interact with intracellular domains of the cytokine receptors and they play crucial roles in transducing signals triggered by the cytokine-receptor interaction. We have previously shown that conditional activation of JAK through membrane-proximal dimerization confers cytokine independence on interleukin-3 (IL-3)-dependent Ba / F3 lymphoid cells and that the cytokine-independent proliferation is completely inhibited by dominant negative Ras. In this work, we demonstrate that ectopic expression of a dominant negative form of Stat5, a major signal transducer and activator of transcription (STAT) expressed in Ba / F3 cells, also inhibits JAK-triggered mitogenesis. In contrast, overexpression of constitutively active Ras or conditional activation of Stat5 by chemical dimerization fails to confer cytokine-independence. However, concomitant activation of ectopic Ras and Stat5 molecules in Ba / F3 cells suffices for cell proliferation in the absence of IL-3. Our results indicate that Ras and STAT are essential and sufficient components of JAK-triggered mitogenesis. Our findings further indicate that the cytokine signal bifurcates into Ras and STAT pathways following JAK activation. PMID- 10835499 TI - Enhancing effect of an inhibitor of nitric oxide synthesis on bacillus Calmette Guerin-induced macrophage cytotoxicity against murine bladder cancer cell line MBT-2 in vitro. AB - We studied the effect of an inhibitor of nitric oxide (NO) synthesis, N(G) monomethyl-L-arginine (L-NMMA), on the Bacillus Calmette-Guerin (BCG)-induced antitumor activity of murine peritoneal exudate cells (PEC) against murine bladder cancer cell line MBT-2 in vitro. L-NMMA enhanced BCG-induced cytotoxic activity of PEC, as well as interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha production. The L-NMMA-induced enhancement was due to the prolonged survival of BCG in macrophages, because no enhancement of cytotoxicity was observed and neither IFN-gamma nor TNF-alpha production was significantly enhanced by killed BCG. Anti-TNF-alpha antibody (Ab) and anti-IFN-gammaAb reduced the L-NMMA-induced enhancement of the cytotoxicity. The depletion of T cells from PEC reduced the production of both IFN-gamma and TNF-alpha, as well as the enhancement of cytotoxicity induced by viable BCG plus L-NMMA. These results suggest that L-NMMA has an enhancing effect on BCG-induced macrophage cytotoxicity and the enhancement is partially mediated by T cells and their soluble products. Accordingly, NO inhibitor should be a valuable adjunct to BCG immunotherapy for bladder cancer. PMID- 10835500 TI - Establishment and characterization of 6-[[2-(Dimethylamino)ethyl]amino]-3-hydroxy 7H-indeno[2,1-c]quinolin-7-one dihydrochloride (TAS-103)-resistant cell lines. AB - 6-2-(Dimethylamino)ethylamino-3-hydroxy-7H-indeno2, 1-cquinolin-7-one dihydrochloride (TAS-103) is a novel anticancer agent that was developed to target both topoisomerase (Topo) I and Topo II. To elucidate its mechanism of action, we have established and characterized TAS-103-resistant cells, derived from mouse leukemia (P388), human colon cancer (DLD-1), and human lung adenocarcinoma (A549) cell lines, by exposure to stepwisely increasing concentrations of TAS-103 in the culture medium. P388 / TAS cells showed only cross-resistance to VP-16 and adriamycin (ADR). The Topo II activity in these cells was decreased to below one-fourth of that in the parental cells, while the Topo I activity remained unchanged. DLD / TAS cells appeared to be cross resistant to VP-16, ADR, camptothecin (CPT), SN-38 and vincristine (VCR). The enzymatic activities of both Topo I and Topo II in these cells were decreased to one-fourth of that observed in the parental cells. Furthermore, the decreased activities were accompanied by lower expression at the mRNA and protein levels. A549 / TAS cells acquired cross-resistance to VP-16, ADR and VCR, though the Topo activities were virtually unchanged. In this cell line, the intracellular accumulation of TAS-103 was significantly decreased and the expression of multidrug resistance associated protein (MRP) was elevated when compared with the parental cells. The results indicate that the affected activities of Topo I and / or Topo II, and in some instances decreased accumulation of TAS-103, are associated with the development of resistance to TAS-103, although the main mechanism of resistance to TAS-103 varied among cell lines. PMID- 10835501 TI - Identification and characterization of a deletion mutant of DNA topoisomerase I mRNA in a camptothecin-resistant subline of human colon carcinoma. AB - In previous studies, we established two camptothecin (CPT)-resistant sublines, HT 29 / CPT and St-4 / CPT, from the human colon cancer cell line HT-29 and the human stomach cancer cell line St-4, respectively. Cellular contents of DNA topoisomerase I (topo I) in the resistant cells were eight-fold less than those in the corresponding parental lines. In this study, we have shown expression of two species of the TOP1 mRNA in HT-29 / CPT. The longer mRNA (4.0 kb) is the wild type TOP1 mRNA, and the shorter mRNA (3.3 kb) proved to have a deletion of 672 bp (nucleotides 58 - 729 or 59 - 730) that caused the in-frame deletion of amino acids 20 - 243 of human topo I. The deleted region is identical to exons 3 - 9 of the TOP1 gene. The expression level of the 3.3-kb mRNA was similar to that of the wild-type mRNA in HT-29 / CPT. St-4 / CPT expressed only the wild-type TOP1 mRNA in lesser amounts than did St-4. Mouse NIH3T3 cells transfected with the wild type TOP1 cDNA showed higher sensitivity to CPT than the parental cells, whereas those transfected with the deleted TOP1 cDNA showed levels similar to those of the parental cells. Expression of the exogenous TOP1 mRNA was confirmed; however, expression of the truncated topo I was not detected in cells transfected with the deleted TOP1 cDNA. These results suggest that the expression of the deleted TOP1 mRNA led to the low expression of CPT-sensitive topo I in the resistant cells. PMID- 10835502 TI - Effects of photodynamic therapy using mono-L-aspartyl chlorin e6 on vessels and its contribution to the antitumor effect. AB - The effect of photodynamic therapy (PDT) on the vascular system has a significant role in tumor tissue destruction. We investigated the contribution of vascular damage to the antitumor effects of PDT and analyzed the quantitative vascular changes after PDT. Fibrosarcoma-bearing BALB / c male mice were injected with mono-L-aspartyl chlorin e6 (NPe6) at a dose of 0.25, 5 or 15 mg / kg, and photoradiation was performed with a diode laser 10 min, 2 h or 24 h after injection, respectively. Ten minutes after injection of 0. 25 mg / kg, NPe6 was found to be present only in plasma, while at 2 h after injection of 5 mg / kg it was present in both plasma and tumor, and 24 h after injection of 15 mg / kg it was present only in the tumor. The antitumor effects observed in the 5 mg / kg-2 h and 0. 25 mg / kg-10 min groups were virtually the same, whereas the effect in the 15 mg / kg-24 h group was weaker. The damage to the tumor vasculature and tumor cells in the 15 mg / kg-24 h group occurred later than under the other conditions, and vascular damage in the tumor-surrounding tissue was also less marked even 24 h after PDT. These results suggested that the plasma NPe6 concentration during laser irradiation contributed more than the tumor NPe6 concentration to the antitumor effect, and that the minimal damage to blood vessels around the tumor at the low plasma NPe6 concentration may be one reason for the failure to obtain a marked antitumor effect. PMID- 10835503 TI - Usefulness of tirapazamine as a combined agent in chemoradiation and thermo chemoradiation therapy at mild temperatures: reference to the effect on intratumor quiescent cells. AB - C3H / He mice bearing SCC VII tumors received 5-bromo-2'-deoxyuridine (BrdU) continuously for 5 days via implanted mini-osmotic pumps to label all proliferating (P) cells. The mice then received one of six different DNA-damaging agents with or without mild temperature hyperthermia (40 degrees C, 30 min, MTH). These agents were adriamycin (ADM), mitomycin C (MMC), cyclophosphamide (CPA), bleomycin (BLM), cisplatin (CDDP), and tirapazamine (TPZ). After the drug treatment, the tumor-bearing mice were irradiated with a series of doses of gamma rays. Immediately after irradiation, the tumors were excised, minced and trypsinized. The tumor cell suspensions thus obtained were incubated with cytochalasin-B (a cytokinesis blocker), and the micronucleus (MN) frequency in cells without BrdU labeling ( = quiescent (Q) cells) was determined using immunofluorescence staining for BrdU. The MN frequency in the total (P + Q) tumor cells was determined from the tumors that had not been pretreated with BrdU. MTH significantly increased the MN frequency of total cells in tumors irradiated with gamma-rays combined with CPA, BLM, CDDP or TPZ, and that of Q cells in tumors irradiated with gamma-rays combined with BLM or TPZ. The sensitivity difference in the MN frequency between total and Q tumor cells was significantly decreased by the combination with TPZ. TPZ combined with radiotherapy and TPZ combined with thermo-radiotherapy at mild temperatures appear to be promising modalities for sensitizing tumor cells in vivo, including Q tumor cells. PMID- 10835504 TI - Enhanced efficacy of radioimmunotherapy combined with systemic chemotherapy and local hyperthermia in xenograft model. AB - We previously found that the efficacy of radioimmunotherapy (RIT) with (131)I-A7, an IgG(1) against M(r) 45000 glycoprotein on colon cancer, was enhanced by local hyperthermia (HT) or chemotherapy with 5-fluorouracil (5-FU). In this study, we aimed to further enhance its efficacy by combining these three modalities. Human colon cancer xenografts (146 x 12 mm(3)) in Balb / c nu / nu female mice were treated with 9.25 MBq (131)I-A7 i.v. combined with HT (43 degrees C for 1 h) and 5-FU (30 mg / kg / day i.p. for 5 days). Tumor growth delay, (Tq(treated) - Tq(control) )/ Tq(control) where Tq is tumor quadrupling time, in mice treated with RIT + HT + 5-FU was improved to 12.7 from 5.90, 7.55 and 10.1 with RIT alone, RIT + 5-FU and RIT + HT, respectively. Complete response was observed in 4 out of 8 tumors with RIT + HT + 5-FU and 3 out of 10 with RIT + HT. No tumor showed complete response with RIT + 5-FU or RIT alone. 5-FU slightly increased myelotoxicity of RIT, but HT did not affect it. Body weight loss was not enhanced by the combination. These results indicate that the combination of three modalities is a feasible approach to enhance the antitumor efficacy of RIT without serious increase of toxicity. PMID- 10835505 TI - Complex vascular lesions at autopsy in a patient with phentermine-fenfluramin. PMID- 10835509 TI - Is it time to revisit the classification system for cervicovaginal cytology? PMID- 10835507 TI - Apoptosis in HL-60 cells and topoisomerase I in vivo. PMID- 10835510 TI - The sign of Babinski. PMID- 10835513 TI - Comparative genomic analyses of primary effusion lymphoma. AB - BACKGROUND: A rare subset of human immunodeficiency virus (HIV) lymphomas, known as primary effusion lymphomas (PELs), are high-grade tumors carrying human herpes virus 8. Mechanisms postulated to contribute to lymphomagenesis include impaired immune surveillance, alterations in hemopoietic regulatory pathways due to expressed viral genes, and acquisition of genomic alterations in regions of the genome that contain regulatory genes. In PEL, limited information exists about the nature of genome-wide aberrations in these rare lymphomas. METHODS: We used comparative genomic hybridization to detect regions of sequence gain and loss throughout the genome of 8 PEL cases. Regions of DNA sequence loss or gain were confirmed using forward and reverse hybridization and t-statistic analyses. RESULTS: Genomic aberrations were identified in 6 of 8 cases, including recurrent gain of sequence in chromosomes 12 [ish enh (12q22;12q23, 12q12;12q23)] in 3 of 8 cases and X [ish enh (X, Xp)] in 2 of 8 cases. CONCLUSIONS: DNA copy number changes occurred in a majority of PEL cases and are consistent with changes observed in other HIV lymphomas. These observations suggest that common genetic events may occur in HIV-associated lymphoid malignancies, but they probably do not contribute to the unique markers and morphology of PEL. Although individual genetic loci have been evaluated previously in a few PEL cases, to our knowledge this study represents the first reported genome-wide scan of copy number changes in these rare HIV-associated tumors. PMID- 10835514 TI - Is apoptosis a diagnostic marker of acute myocardial infarction? AB - OBJECTIVE: To investigate and quantify the presence of apoptosis in early myocardial ischemia in humans. METHODS: Histologic sections from the left and right ventricles of 16 hearts with impending myocardial infarction were stained with terminal deoxynucleotidyl transferase-mediated deoxyuridine 5-triphosphate nick end labeling (TUNEL) method and with antibodies to p53, bcl-2, cpp32, FAS, FAS-L, and bax. DNA electrophoretic analysis was also performed. RESULTS: According to the inclusion criteria, all 16 cases showed morphologic changes consistent with ischemia and/or reperfusion. TUNEL results were positive in 14 of the 16 ischemic areas. Unexpectedly, they were also positive in "remote from ischemia" myocardium of both the left and right ventricles. DNA electrophoretic analysis confirmed the results of TUNEL. Immunohistochemistry was uniformly negative, probably because of autolysis phenomena. CONCLUSIONS: We showed that apoptosis precedes necrosis in humans, but the detection of apoptosis cannot be used as a diagnostic tool, since it can also be triggered by nonischemic events. PMID- 10835515 TI - Primary nodal gastrinomas. AB - BACKGROUND: The term primary lymph node gastrinoma was first used to describe a group of patients with gastrin-producing tumors present in lymph nodes located in a well-defined anatomic region. The patients had no known primary tumors in the pancreas or gastrointestinal tract and had disease-free survival for up to 18 years. The anatomic region in question has a triangular shape that extends from the cystic and common bile ducts to the second and third portion of the duodenum and the neck and body of the pancreas. The term gastrinoma triangle was coined to identify the area; in addition, it was postulated that lymph nodes located in the gastrinoma triangle normally contained neuroendocrine cells capable of secreting gastrin and other neuropeptides. From its inception, the postulate became the subject of controversy. DESIGN: To extend previous observations, we examined the lymph nodes located in the gastrinoma triangle of 20 autopsy cases for the presence of neuroendocrine cells, as determined by immunohistochemistry, using antibodies to a panneuroendocrine substance (eg, synaptophysin) and a specific neuropeptide (eg, gastrin). Scanning for positive cells was performed by 2 observers (M.E.H. and M.C.C.). We compared the findings in these lymph nodes with lymph nodes obtained from axillary and inguinal dissections during surgical procedures. RESULTS: In all, 417 lymph nodes were studied. Five of the 20 gastrinoma triangle cases contained synaptophysin reactive cells, whereas 3 had gastrin reactive cells. None of the axillary and inguinal lymph nodes contained neuroendocrine cells. CONCLUSION: Our findings support the hypothesis of entrapment of neuroendocrine cells during development and the presence of primary nodal gastrinomas. PMID- 10835516 TI - ras gene mutations in salivary gland tumors. AB - OBJECTIVE: To assess the prevalence of activating mutations in K-ras and H-ras genes in salivary gland tumors with ductal or acinar differentiation and to evaluate their potential correlation with clinical parameters. DESIGN: Paraffin embedded tissue samples of salivary gland carcinomas were investigated by the application of a direct sequence analysis procedure with automated DNA sequencing of polymerase chain reaction-amplified ras sequences. SETTING: Tertiary care teaching hospital. PATIENTS: Twenty-four patients with salivary gland carcinoma were surgically treated. Nine had adenocarcinoma, 1 had adenosquamous carcinoma, 11 had mucoepidermoid carcinoma, and 3 had acinic cell carcinoma. RESULTS: Point mutations were detected in 7 (29%) of the 24 carcinomas examined. The K-ras gene was mutated in only 2 samples (8%): a GGC-to-ATC mutation at codon 13 in an adenocarcinoma and a GGC-to-GTC transversion mutation at codon 13 in a mucoepidermoid carcinoma. Five (21%) harbored H-ras mutations: 4 contained a GGC to-GTC transversion mutation at codon 12 and 1 had 2 distinct mutations, the same G-to-T at codon 12 as was shown in the other cases and a GGT-to-GGA heterozygous mutation at codon 13. All the H-ras mutations were in the group of mucoepidermoid carcinoma lesions (45%; 5/11). CONCLUSION: Our data suggest that K-ras gene alteration is probably not an important factor in the oncogenesis of human salivary gland tumors. However, mutational activation of the H-ras gene appears to play a role in the development and/or progression of salivary gland mucoepidermoid carcinomas. PMID- 10835517 TI - Comparison of the effectiveness of polymerase chain reaction and enzyme immunoassay in detecting Chlamydia trachomatis in different female genitourinary specimens. AB - BACKGROUND: In high-volume laboratories, enzyme immunoassay (EIA) is the most commonly used method of detecting Chlamydia trachomatis. The optimal specimen for detecting C trachomatis is a combined urethral and cervical swab. OBJECTIVE: To compare EIA with the combined urethral and cervical swab with polymerase chain reaction (PCR) on urine alone and urine mixed with cervical cells. PATIENTS AND METHODS: Phase 1 of the study included 752 sets of specimens used for comparison. In phase 2, another 212 samples of urine and urine plus cervical cells were added to the study for comparison of the 2 specimen types using PCR. RESULTS: In phase 1, 648 samples were negative and 76 were positive by all 3 methods and specimen combinations. Enzyme immunoassay was able to detect 81 positive samples (10.8%), whereas PCR on urine alone detected 97 positive samples (12.9%) and PCR on urine plus cervical cells detected 102 positive samples (13.6%), giving a sensitivity of 75%, 93.3%, and 98. 1% respectively. In phase 2, PCR on urine alone detected 119 positive samples (12.3%) and PCR on urine plus cervical cells detected 127 positive samples (13.1%), with a sensitivity of 92.2% and 98.5%, respectively. CONCLUSION: Polymerase chain reaction on urine alone or urine plus cervical cells is superior to EIA on combined cervical and urethral swabs. There is a slight advantage of adding cervical cells to the urine compared with the urine specimen alone when PCR is used as the assay for detection. The total inhibition rate in our female population is only 3.1% when PCR is used. PMID- 10835518 TI - Gynecomastia-like lesions in the female breast. AB - CONTEXT: Gynecomastia is an abnormal enlargement of the male breast, but the histopathologic abnormalities could theoretically be present in female breasts as well. To my knowledge, however, there have been no reports in the literature to date that have attributed a clinically or radiologically detectable mass in a female breast exclusively to the histopathologic findings seen in gynecomastia. OBJECTIVE: The clinical, radiologic, and histopathologic findings of 4 patients with lesions composed exclusively of the findings of gynecomastia, herein termed gynecomastia-like lesions, are presented and compared with those of 3 patients with lesser degrees of these changes, herein termed gynecomastia-like areas, which were incidental accompaniments to other lesions. SETTING: During a 26-month interval, 1242 breast excisions (excluding needle biopsies or aspirates) were examined in a 170-bed, acute-care, community-medical school teaching hospital. RESULTS: Four patients had gynecomastia-like lesions varying from 1 to 3 cm in greatest dimension with histopathologic features showing the changes of gynecomastia exclusively. Two patients had clinically palpable masses. Two other patients had masses detected only by mammography. One postmenopausal patient was taking estrogen/progesterone replacement medication. Two of the 3 others were not taking birth control pills, and none had a clinical endocrinopathy. Thus far, none of the lesions have recurred. The 3 patients with gynecomastia-like areas were diagnosed separately with infiltrating ductal carcinoma, fibroadenoma, and lipoma. CONCLUSIONS: Pathologists should be aware that clinically palpable or radiologically detected masses in female breasts may be composed exclusively of the histopathologic findings of gynecomastia. PMID- 10835519 TI - Increased midkine expression in hepatocellular carcinoma. AB - CONTEXT: Midkine (MK) is a novel heparin-binding growth factor whose gene was identified in embryonal carcinoma cells in early stages of retinoic acid-induced differentiation. OBJECTIVE: To examine the overexpression of MK in hepatocellular carcinoma (HCC). METHODS: Seventy-seven primary HCC specimens from patients aged 17 to 72 years (63 men and 14 women) were examined. Histologically, 16 cases of HCC were classified as the well-differentiated type, 50 cases as the moderately differentiated type, and 11 cases as the poorly differentiated type. Immunohistochemical analysis was performed using a rat immunoglobulin G2a monoclonal antibody against the carboxyl terminal region of human MK. In situ hybridization was also performed on 20 HCC samples. RESULTS: We successfully applied this monoclonal antibody against MK to analyze archival paraffin sections. The cancer tissues showed a positive reaction to this antibody, in which there was an intense reaction in their cytoplasm. Approximately one third of the individuals with HCC (26/77) had tumor cells that expressed MK, and these were classified into the following types: moderately differentiated (20/50), well differentiated (3/16), and poorly differentiated (3/11). The in situ hybridization analysis revealed that the signals of MK transcripts were found in the cytoplasm of the cancer cells; the distribution and localization of the MK transcripts' signals determined by in situ hybridization analysis were similar to those obtained by immunohistochemical analysis. CONCLUSIONS: Hepatocellular carcinoma expressed increased MK at the messenger RNA and protein level. PMID- 10835520 TI - Informed evaluation of pathology residency programs. AB - OBJECTIVES: To identify resources and summarize important issues in anatomic and clinical pathology training and to assist the pathology resident candidate in evaluating potential training programs. DATA SOURCES: Published guides for medical residency applicants, recent literature discussing pathology education, and World Wide Web sites. STUDY SELECTION: Resources perceived by the author as valuable for the pathology resident candidate. DATA EXTRACTION: Key issues in pathology education are identified. DATA SYNTHESIS: Issues are discussed from the perspective of a pathology resident candidate, and resources for further information are provided. CONCLUSIONS: The pathology residency candidate faces unique challenges in the residency search process because of the breadth of pathology training and the limited exposure to the practice of pathology in medical school. General guides for residency applicants include little discussion of pathology-specific issues. Recent literature discussing pathology education is fragmented but provides invaluable insights for resident candidates. This review seeks to identify a wide variety of issues and resources as a starting point for evaluating potential training programs. PMID- 10835521 TI - Waterhouse-Friderichsen syndrome secondary to Capnocytophaga canimorsus septicemia and demonstration of bacteremia by peripheral blood smear. AB - Waterhouse-Friderichsen syndrome caused by Capnocytophaga canimorsus septicemia was fatal in a previously healthy 47-year-old woman. The patient died suddenly in less than 12 hours after presentation, in spite of supportive measures, including ventilation, antibiotic coverage, pressor therapy, and multiple transfusions of blood products. The diagnosis of infection due to an unusual organism was suspected earlier in the course of management after review of the peripheral blood smear. The importance of the findings in the blood smear and their correlation with infection due to this organism are discussed. PMID- 10835522 TI - Malignant peripheral nerve sheath tumor with a t(X;18). AB - We describe an ankle tumor arising in a 16-year-old girl. The tumor demonstrated histology typical of a malignant peripheral nerve sheath tumor (MPNST), but exhibited a variant form of the (X;18) translocation associated with synovial sarcoma. Immunohistochemical stains were positive for vimentin, CD57, collagen type IV, and Bcl-2. Routine and molecular cytogenetic studies showed an unbalanced 3-way chromosomal translocation that involved chromosomes X, 18, and 1. Electron microscopic findings were noncontributory. This unusual tumor raises the following questions and possibilities: (1) As the t(X;18) suggests, could this tumor be a monophasic synovial sarcoma with the histologic features of an MPNST? (2) Or, as the histology suggests, is this tumor an MPNST that has a t(X;18)? (3) Finally, could MPNST histology, a t(X;18), and no defining immunohistochemical or electron microscopic features represent an as yet unrecognized part of a spectrum that spans from synovial sarcoma to MPNST or other spindle cell tumors? PMID- 10835523 TI - Low-grade intraosseous osteosarcoma with prominent lymphoid infiltrate. AB - Low-grade intraosseous osteosarcoma is an uncommon and well-differentiated osteosarcoma with a good prognosis. We report a proximal tibial low-grade intraosseous osteosarcoma with a prominent intratumoral lymphoid infiltrate, which led to an initial diagnosis of probable malignant lymphoma. The importance of this infiltrate, which exhibited reactive features on flow cytometric studies, is not known. Our patient is free of tumor 1 year after limb salvage surgery, without hematologic or lymphoid abnormalities. PMID- 10835524 TI - The amyloid deposit in calcifying epithelial odontogenic tumor is immunoreactive for cytokeratins. AB - Calcifying epithelial odontogenic tumor, also known as Pindborg tumor, is a rare benign tumor with locally aggressive behavior. It is characterized by squamous epithelial cells, calcifications, and eosinophilic deposits that have been identified as amyloid. We report a case of calcifying epithelial odontogenic tumor and investigate the nature of the amyloid, using histologic, immunohistochemical, and ultrastructural studies. The amyloid was immunohistochemically negative for basement membrane components and positive for all cytokeratin stains performed (cocktail of cytokeratins 1, 5, 6, 8, 13, and 16, and cytokeratins AE1 and AE3). The amyloid stained focally in a glandular like pattern, reminiscent of the epithelial glandlike structures of the tumor. We conclude that the amyloid is derived from filamentous degeneration of keratin filaments that originate from the tumor squamous epithelium. The keratin degeneration is part of a developmental or aging process that the tumor undergoes. PMID- 10835525 TI - Paraganglioma of the tongue. AB - Paragangliomas are neuroendocrine tumors arising from extra-adrenal autonomic ganglia. We present what is to our knowledge the first immunohistochemically documented case of a paraganglioma of the tongue, a 2.5-cm benign tumor growing in the position of the foramen cecum. The patient was an elderly woman who presented with throat irritation. The histologic profile and ultrastructural appearance of this lesion were classic for paraganglioma, and cells stained strongly for neurosecretory granules by immunohistochemistry. Carcinoid, ectopic thyroid neoplasm, and other tumors that mimic paraganglioma were ruled out on immunohistochemical grounds. To our knowledge, paraganglia have not been previously documented in the tongue; this tumor may have arisen from a branch of cranial nerve VII or IX, or from an embryologic remnant of the thyroid or thyroid capsule. PMID- 10835526 TI - Chromosome 22q11 deletion complicated by dissecting pulmonary arterial aneurysm and jejunal atresia in an infant. AB - We present an autopsy case of a 46-day-old male infant with chromosome 22q11 deletion, which is considered the primary cause of several diseases, including DiGeorge syndrome and velocardiofacial syndrome. The patient had 2 notable congenital abnormalities: multiple dissecting pulmonary arterial aneurysms distributed in both lungs and multiple jejunal atresia with apple-peel deformity. The former, a very rare pathologic condition especially in infancy, was found incidentally at autopsy and was the primary cause of death. To our knowledge, neither of these lesions has been reported previously in a patient with chromosome 22q11 deletion. PMID- 10835527 TI - Identification of the asexual state of Rhizopus species on histologic tissue sections in a patient with rhinocerebral mucormycosis. AB - Mucormycosis is an infection caused by a group of fungi in the order Mucorales in the phylum Zygomycota. The most well-known form of this disease is rhinocerebral mucormycosis, which usually develops in diabetic or immunocompromised patients. The fungal hyphal elements are easily detected in biopsy specimens by direct or histologic examination. However, the confirmatory identification of the genus or species requires culture of the specimen. This article presents a case of rhinocerebral mucormycosis in which presumptive identification of the genus was made without microbiologic cultures and was based on the extraordinarily rare appearance of fungal sporangia and sporangiospores in histologic tissue sections. Identification of these structures allowed an early and accurate diagnosis of rhinocerebral invasive mucormycosis. PMID- 10835528 TI - Carcinosarcoma of the liver. AB - An autopsy case of carcinosarcoma of the liver in an 84-year-old man is described. The 14 x 6-cm solid tumor was located in the hilus to the left lobe and was grayish-white with some translucent areas. Histologically, the tumor consisted of an intimate mixture of adenocarcinomatous and chondrosarcomatous elements with transitional areas in between. Immunohistochemically, cells of the adenocarcinomatous elements were positive for cytokeratin but negative for S100 protein, whereas cells of the chondrosarcomatous elements showed the reverse staining pattern. Cells of transitional areas were positive for both cytokeratin and S100 protein. Most previously reported cases of carcinosarcoma of the liver have involved elderly men and have had a poor prognosis. The findings of the present case support the view that carcinosarcomas represent carcinomas that develop a sarcomatous element via metaplasia of the epithelial element. PMID- 10835530 TI - Spindle cell tumor of urinary bladder serosa with phenotypic and genotypic features of gastrointestinal stromal tumor. AB - Most mesenchymal tumors of the gastrointestinal tract are now referred to as gastrointestinal stromal tumors (GISTs). The tumors differ from ordinary leiomyomas and schwannomas in several respects: the GISTs typically express c-kit protein (CD117) and CD34, 30% to 50% of them are (often focally) positive for alpha-smooth muscle actin, and all are negative for desmin and S100 protein. Recently, mutations in the exon 11 of the c-kit gene have been identified and confirmed as a molecular genetic marker for the subset of GISTs. In this report, we describe a mesenchymal tumor removed from the pelvic cavity of a 52-year-old woman, who is alive without disease 36 months after the surgery. The 5-cm tumor was densely attached to the external aspect of the urinary bladder but was attached to small intestine by only filmy adhesions. The tumor grossly resembled a leiomyoma and was histologically composed of sheets of spindle cells with a dense collagenous background. The mitotic activity was low, less then 1 per 50 high-power fields. Immunohistochemically, the tumor cells were negative for alpha smooth muscle actin and desmin and positive for CD117 and CD34. Molecular genetic analysis of the exon 11 of the c-kit gene revealed a point mutation in the region commonly mutated in GISTs. This mutation substituted T for A in the codon 557, leading to the change of amino acid sequence (tryptophan for arginine) of the KIT protein. This case illustrates that tumors phenotypically and genotypically similar to GISTs may present in sites other than the tubular gastrointestinal tract. PMID- 10835529 TI - Waldenstrom macroglobulinemia presenting as a pleural effusion. AB - We describe a patient with Waldenstrom macroglobulinemia who presented with a lymphocytic pleural effusion. Pleural involvement in Waldenstrom macroglobulinemia is very rare. In addition, to our knowledge, there are no reports in the literature in which a primary diagnosis was made on the basis of pleural fluid analysis. The presence of small lymphocytes in serous cavity fluid can pose great difficulty in the differentiation between a low-grade lymphoproliferative disorder and reactive lymphocytosis. The diagnosis of malignancy in this case was established on the basis of morphologic testing, flow cytometry, and the detection of B-cell immunoglobulin gene rearrangement. PMID- 10835531 TI - Meningioma presenting as an intraoral mass in a patient with neurofibromatosis type 1. AB - A 77-year-old woman with neurofibromatosis type 1 presented with ill-fitting dentures due to intraoral extension of a right temporal fossa mass. Computed tomographic scanning demonstrated that the masticator space mass bowed the zygomatic arch and remodeled the lateral orbit and maxillary sinus walls, findings that were consistent with the clinical diagnosis of a neurofibroma with possible malignant transformation. However, light microscopic, immunohistochemical, and ultrastructural examination of tissue from an incisional biopsy specimen were diagnostic of meningioma. This case illustrates that the clinicopathologic differential diagnosis of an enlarging mass in patient with neurofibromatosis should include sporadic, unrelated neoplasms as well as tumors known to be associated with the syndrome. PMID- 10835532 TI - Persistent tuberculosis or specimen contamination? AB - Cross-contamination during sequential processing of sputum specimens from different patients causes false-positive growth of Mycobacterium tuberculosis in culture. We describe an unusual case of cross-contamination in a 36-year-old man with acquired immunodeficiency syndrome and possible persistent tuberculosis. Culture with 1 of 3 sputum specimens was positive for rifampin-susceptible M tuberculosis. Review of processing revealed that his single culture-positive sputum specimen had followed a sputum specimen from another patient with active pulmonary tuberculosis that was positive in culture for M tuberculosis resistant to rifampin. Molecular strain typing by restriction fragment length polymorphism demonstrated the 2 isolates to be an identical strain of M tuberculosis. Agar proportion susceptibility testing of the rifampin-resistant isolate revealed low numbers of resistant organisms in a range of 1.5% to 3.3%. It was concluded that rifampin-susceptible organisms that constituted approximately 98% of the resistant isolate contaminated sputum from the patient with possible persistent tuberculosis. His culture result was, therefore, considered false positive, not an indication of tuberculosis. PMID- 10835533 TI - Variant chromophobe renal cell carcinoma. AB - Separation of renal cell tumors into different prognostic groups is an imperative function of the diagnostic pathologist. Recently, chromophobe renal carcinoma has been described as a tumor that is morphologically distinct from conventional "clear cell" carcinoma and that has a low metastatic potential. Identification is based on routine light microscopic features and is confirmed by special stains, immunohistochemistry, and electron microscopy. We present a variant of chromophobe renal carcinoma that did not show the typical cytomorphologic features on light microscopy after formaldehyde fixation. After fixation in Solufix (a commercial fixative), these features were recognized and the diagnosis was confirmed. The tumor also showed an unusual form of calcification and psammoma body formation not previously recognized in chromophobe tumors. Molecular biological assessment was inconclusive, but excluded a chromosome 3p deletion usually found in conventional renal carcinoma. The use of a different primary fixative may provide a cost-effective screening tool to detect variant renal tumors and may have important prognostic implications. PMID- 10835534 TI - Composite prolymphocytoid and hodgkin transformation of chronic lymphocytic leukemia. AB - The indolent course of chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL) is occasionally altered by transformation to a histologically distinct, rapidly progressive, and clinically unresponsive hematologic malignant neoplasm. We report a case of CLL that, after 3 years of slowly progressive disease and treatment with single-agent chemotherapy (fludarabine phosphate), underwent a composite prolymphocytoid and classic Hodgkin lymphoma transformation. The diagnosis of classic Hodgkin lymphoma was based on the presence of Reed-Sternberg cells with typical morphologic structure and immunophenotype (CD15(+), CD30(+), CD45(-), CD20(-)) associated with the characteristic polymorphous inflammatory background consisting of numerous eosinophils, plasma cells, and reactive T lymphocytes. The remainder of the lymph node and the peripheral blood showed increased numbers of prolymphocytes admixed with typical small CLL cells. Recognition of such a transformation is of the utmost importance, since histologically similar Reed-Sternberg-like cells may be seen in Richter transformation. In contrast to prolymphocytoid transformation of CLL, Richter syndrome is rapidly fatal, with a median survival of 4 to 5 months. The patient pursued a clinical course similar to pure prolymphocytoid transformation and died with disease after 30 months following treatment with combination chemotherapy. PMID- 10835536 TI - Pathologic quiz case. A 71-year-old man with recurrent gross hematuria. PMID- 10835535 TI - Multiple myeloma involving the myocardium and coronary vessels. AB - We report a case of metastatic plasmacytoma to the myocardium and coronary vessels in a 57-year-old man with multiple myeloma. Originally, the patient had a large plasmacytoma in his left chest wall and lung. He received local radiation and chemotherapy. Subsequently, the patient presented with symptoms of congestive heart failure. He had no prior history of cardiac disease. The patient was treated medically and later died from respiratory failure. At autopsy, a metastatic plasmacytoma was identified within the myocardium and externally compressing the coronary arteries. The tumor infiltrated into the coronary sinus. It is difficult to speculate whether the patient's symptoms were due to cardiac involvement since the tumor burden in his chest was also considerable. To our knowledge, coronary vessel involvement with plasmacytoma has not been previously described. PMID- 10835538 TI - Pathologic quiz case. Fourteen-year-old with painful lytic lesion of the tibia. PMID- 10835537 TI - Pathologic quiz case. Abdominal mass with adhesions. PMID- 10835540 TI - Human anti-mouse antibodies. AB - Human anti-mouse antibodies (HAMA) are human immunoglobulins with specificity for mouse immunoglobulins. This topic currently is of interest because of the increased use of monoclonal mouse antibodies as diagnostic reagents both for in vitro laboratory measurements and for in vivo imaging studies. Monoclonal mouse antibodies also are being used therapeutically. This short article reviews the production of HAMA in patients receiving monoclonal antibodies and illustrates the potential ways that HAMA can interfere with immunoassay measurements. Methods for measuring and neutralizing HAMA also are discussed. PMID- 10835539 TI - Pathologic quiz case. A unilateral renal mass in an elderly woman. PMID- 10835541 TI - Mitral annular calcification with Staphylococcus aureus periannular abscess. PMID- 10835544 TI - Atlas of renal pathology PMID- 10835542 TI - Ticked off: Ixodes. PMID- 10835545 TI - Stemming doublespeak. PMID- 10835546 TI - In praise of pessimism. PMID- 10835547 TI - Organization becomes cause in the matter. PMID- 10835548 TI - How important is genetic engineering to European seed firms? PMID- 10835549 TI - Total RNA and array-based expression monitoring. PMID- 10835550 TI - Reply to 'Total RNA and array-based expression monitoring' PMID- 10835551 TI - Integrated pararetroviral sequences. PMID- 10835553 TI - Japan's cloning law fails PMID- 10835554 TI - Cartoon PMID- 10835552 TI - Gene therapy guidelines PMID- 10835556 TI - GMO roundup PMID- 10835555 TI - Structural genomics deal PMID- 10835557 TI - Ruling favors amgen PMID- 10835558 TI - EU's new orphan drug regulations PMID- 10835559 TI - India okays Bt cotton PMID- 10835561 TI - Human fly bites rub celera PMID- 10835560 TI - EU biosciences panel PMID- 10835562 TI - EPA rejects greenpeace PMID- 10835564 TI - Research collaborations PMID- 10835563 TI - Eco-terrorism penalties PMID- 10835566 TI - US house biotech report PMID- 10835565 TI - Bioprospecting deal OK PMID- 10835567 TI - Stem cell hearing PMID- 10835568 TI - Cold fusion for eggs PMID- 10835569 TI - Soluble receptor against lupus? PMID- 10835570 TI - Vaccinia's interactions PMID- 10835571 TI - High-affinity TCRs PMID- 10835572 TI - Surrogate beta-cells PMID- 10835573 TI - In vivo correction of muscular dystrophy PMID- 10835574 TI - Metallothionein biomediator PMID- 10835575 TI - Surface vaccine combo PMID- 10835576 TI - Engineering heparin-independent FGF PMID- 10835578 TI - Targeting gene therapy to airway epithelia PMID- 10835577 TI - Massively massive parallel analysis PMID- 10835580 TI - Reconstituting active G-coupled receptors on magnetic beads PMID- 10835581 TI - Imaging neuronal activity in small animals PMID- 10835579 TI - Technical reports PMID- 10835582 TI - Retroviral vector for quiescent cells? PMID- 10835583 TI - Review PMID- 10835584 TI - Healthier ketchup? PMID- 10835585 TI - Industry still fears political opposition to european union GM legislation PMID- 10835586 TI - Lukewarm reception for US biotech proposals PMID- 10835587 TI - Local efforts to enforce GM food labeling are thwarted PMID- 10835589 TI - Small success for gene therapy PMID- 10835588 TI - IPOs, not follow-ons, proceed PMID- 10835590 TI - Cloned cows turn back the cellular clock. PMID- 10835591 TI - Protein biopesticide may be next wave in pest control. PMID- 10835592 TI - Taking a census of mRNA populations with microbeads. PMID- 10835593 TI - Laying hold on eternal life? PMID- 10835594 TI - Microbes with a mettle for bioremediation. PMID- 10835595 TI - Silencing Morpheus awakens transgenes. PMID- 10835596 TI - European exploitation of biotechnology-do government policies help? A recent survey of public spending on biotechnology in Europe suggests that money alone cannot stimulate growth of the sector. PMID- 10835597 TI - Who's your neighbor? New computational approaches for functional genomics. AB - Several recently developed computational approaches in comparative genomics go beyond sequence comparison. By analyzing phylogenetic profiles of protein families, domain fusions, gene adjacency in genomes, and expression patterns, these methods predict many functional interactions between proteins and help deduce specific functions for numerous proteins. Although some of the resultant predictions may not be highly specific, these developments herald a new era in genomics in which the benefits of comparative analysis of the rapidly growing collection of complete genomes will become increasingly obvious. PMID- 10835598 TI - In vivo targeted repair of a point mutation in the canine dystrophin gene by a chimeric RNA/DNA oligonucleotide. AB - In the canine model of Duchenne muscular dystrophy in golden retrievers (GRMD), a point mutation within the splice acceptor site of intron 6 leads to deletion of exon 7 from the dystrophin mRNA, and the consequent frameshift causes early termination of translation. We have designed a DNA and RNA chimeric oligonucleotide to induce host cell mismatch repair mechanisms and correct the chromosomal mutation to wild type. Direct skeletal muscle injection of the chimeric oligonucleotide into the cranial tibialis compartment of a six-week-old affected male dog, and subsequent analysis of biopsy and necropsy samples, demonstrated in vivo repair of the GRMD mutation that was sustained for 48 weeks. Reverse transcription-polymerase chain reaction (RT-PCR) analysis of exons 5-10 demonstrated increasing levels of exon 7 inclusion with time. An isolated exon 7 specific dystrophin antibody confirmed synthesis of normal-sized dystrophin product and positive localization to the sarcolemma. Chromosomal repair in muscle tissue was confirmed by restriction fragment length polymorphism (RFLP)-PCR and sequencing the PCR product. This work provides evidence for the long-term repair of a specific dystrophin point mutation in muscle of a live animal using a chimeric oligonucleotide. PMID- 10835599 TI - Spleen necrosis virus-derived C-type retroviral vectors for gene transfer to quiescent cells. AB - Gene therapy applications of retroviral vectors derived from C-type retroviruses have been limited to introducing genes into dividing target cells. Here, we report genetically engineered C-type retroviral vectors derived from spleen necrosis virus (SNV), which are capable of infecting nondividing cells. This has been achieved by introducing a nuclear localization signal (NLS) sequence into the matrix protein (MA) of SNV by site-directed mutagenesis. This increased the efficiency of infecting nondividing cells and was sufficient to endow the virus with the capability to efficiently infect growth-arrested human T lymphocytes and quiescent primary monocyte-derived macrophages. We demonstrate that this vector actively penetrates the nucleus of a target cell, and has potential use as a gene therapy vector to transfer genes into nondividing cells. PMID- 10835600 TI - Gene expression analysis by massively parallel signature sequencing (MPSS) on microbead arrays. AB - We describe a novel sequencing approach that combines non-gel-based signature sequencing with in vitro cloning of millions of templates on separate 5 microm diameter microbeads. After constructing a microbead library of DNA templates by in vitro cloning, we assembled a planar array of a million template-containing microbeads in a flow cell at a density greater than 3x10(6) microbeads/cm2. Sequences of the free ends of the cloned templates on each microbead were then simultaneously analyzed using a fluorescence-based signature sequencing method that does not require DNA fragment separation. Signature sequences of 16-20 bases were obtained by repeated cycles of enzymatic cleavage with a type IIs restriction endonuclease, adaptor ligation, and sequence interrogation by encoded hybridization probes. The approach was validated by sequencing over 269,000 signatures from two cDNA libraries constructed from a fully sequenced strain of Saccharomyces cerevisiae, and by measuring gene expression levels in the human cell line THP-1. The approach provides an unprecedented depth of analysis permitting application of powerful statistical techniques for discovery of functional relationships among genes, whether known or unknown beforehand, or whether expressed at high or very low levels. PMID- 10835601 TI - G-protein-coupled receptors as targets for gene transfer vectors using natural small-molecule ligands. AB - Gene therapy for cystic fibrosis (CF) has focused on correcting electrolyte transport in airway epithelia. However, success has been limited by the failure of vectors to attach and enter into airway epithelia, and may require redirecting vectors to targets on the apical membrane of airway cells that mediate these functions. The G-protein-coupled P2Y2 receptor (P2Y2-R) is abundantly expressed on the airway lumenal surface and internalizes into coated pits upon agonist activation. We tested whether a small-molecule-agonist (UTP) could direct vectors to P2Y2-R and mediate attachment, internalization, and gene transfer. Fluorescein UTP studies demonstrated that P2Y2-R agonists internalized with their receptor, and biotinylated UTP (BUTP) mediated P2Y2-R-specific internalization of fluorescently labeled streptavidin (SAF) or SAF conjugated to biotinylated Cy3 adenoviral-vector (BCAV). BUTP conjugated to BCAV mediated P2Y2-R-specific gene transfer in (1) adenoviral-resistant A9 and polarized MDCK cells by means of heterologous P2Y2-R, and (2) well-differentiated human airway epithelial cells by means of endogenous P2Y2-R. Targeting vectors with small-molecule-ligands to apical membrane G-protein-coupled receptors may be a feasible approach for successful CF gene therapy. PMID- 10835602 TI - Engineering of an FGF-proteoglycan fusion protein with heparin-independent, mitogenic activity. AB - In the absence of heparan sulfate (HS) on the surface of target cells, or free heparin (HP) in the vicinity of their receptors, fibroblast growth factor (FGF) family members cannot exert their biological activity and are easily damaged by proteolysis. This limits the utility of FGFs in a variety of applications including treatment of surgical, burn, and periodontal tissue wounds, gastric ulcers, segmental bony defects, ligament and spinal cord injury. Here we describe an FGF analog engineered to overcome this limitation by fusing FGF-1 with HS proteoglycan (PG) core protein. The fusion protein (PG-FGF-1), which was expressed in Chinese hamster ovary cells and collected from the conditioned medium, possessed both HS and chondroitin sulfate sugar chains. After fractionation, intact PG-FGF-1 proteins with little affinity to immobilized HP and high-level HS modification, but not their heparitinase or heparinase digests, exerted mitogenic activity independent of exogenous HP toward HS-free Ba/F3 transfectants expressing FGF receptor. Although PG-FGF-1 was resistant to tryptic digestion, its physiological degradation with a combination of heparitinase and trypsin augmented its mitogenic activity toward human endothelial cells. The same treatment abolished the activity of simple FGF-1 protein. By constructing a biologically active proteoglycan-FGF-1 fusion protein, we have demonstrated an approach that may prove effective for engineering not only FGF family members, but other HP-binding molecules as well. PMID- 10835603 TI - Surface-displayed viral antigens on Salmonella carrier vaccine. AB - We have developed a recombinant live oral vaccine using the ice-nucleation protein (Inp) from Pseudomonas syringae to display viral antigens on the surface of Salmonella spp. Fusion proteins containing viral antigens were expressed in the oral vaccine strain, Salmonella typhi Ty21a. Surface localization was verified by immunoblotting and fluorescence-activated cell sorting. The immunogenicity of surface-displayed viral antigens on the recombinant live vaccine strain was assessed in mice inoculated intranasally and intraperitoneally. Inoculation resulted in significantly higher serum antibody level than those induced by viral antigens expressed intracellularly. Thus, this multivalent mucosal live vaccine may provide an effective means for inducing mucosal or systemic immune responses against multiple viral antigens. PMID- 10835604 TI - Paramagnetic proteoliposomes containing a pure, native, and oriented seven transmembrane segment protein, CCR5. AB - Seven-transmembrane segment, G protein-coupled receptors play central roles in a wide range of biological processes, but their characterization has been hindered by the difficulty of obtaining homogeneous preparations of native protein. We have created paramagnetic proteoliposomes containing pure and oriented CCR5, a seven-transmembrane segment protein that serves as the principal coreceptor for human immunodeficiency virus (HIV-1). The CCR5 proteoliposomes bind the HIV-1 gp120 envelope glycoprotein and conformation-dependent antibodies against CCR5. The binding of gp120 was enhanced by a soluble form of the other HIV-1 receptor, CD4, but did not require additional cellular proteins. Paramagnetic proteoliposomes are uniform in size, stable in a broad range of salt concentrations and pH, and can be used in FACS and competition assays typically applied to cells. Integral membrane proteins can be inserted in either orientation into the liposomal membrane. The magnetic properties of these proteoliposomes facilitate rapid buffer exchange useful in multiple applications. As an example, the CCR5-proteoliposomes were used to select CCR5-specific antibodies from a recombinant phage display library. Thus, paramagnetic proteoliposomes should be useful tools in the analysis of membrane protein interactions with extracellular and intracellular ligands, particularly in establishing screens for inhibitors. PMID- 10835605 TI - In vivo imaging of neuronal activation and plasticity in the rat brain by high resolution positron emission tomography (microPET). AB - The study of neural repair and neuroplasticity in rodents would be enhanced by the ability to assess neuronal function in vivo. Positron emission tomography (PET) is used to study brain plasticity in humans, but the limited resolution and sensitivity of conventional scanners have generally precluded the use of PET to study neuroplasticity in rodents. We now demonstrate that microPET, a PET scanner developed for use with small animals, can be used to assess metabolic activity in different regions of the conscious rodent brain using [18F]fluorodeoxyglucose (FDG) as the tracer, and to monitor changes in neuronal activity. Limbic seizures result in dramatically elevated metabolic activity in the hippocampus, whereas vibrissal stimulation results in more modest increases in FDG uptake in the contralateral neocortex. We also show that microPET can be used to study lesion induced plasticity of the brain. Cerebral hemidecortication resulted in diminished relative glucose metabolism in the neostriatum and thalamus ipsilateral to the lesion, with subsequent, significant recovery of metabolic function. These studies demonstrate that microPET can be used for serial assessment of metabolic function of individual, awake rats with a minimal degree of invasiveness, and therefore, has the potential for use in the study of brain disorders and repair. PMID- 10835606 TI - Engineering a mouse metallothionein on the cell surface of Ralstonia eutropha CH34 for immobilization of heavy metals in soil. AB - Here we describe targeting of the mouse metallothionein I (MT) protein to the cell surface of the heavy metal-tolerant Ralstonia eutropha (formerly Alcaligenes eutrophus) CH34 strain, which is adapted to thrive in soils highly polluted with metal ions. DNA sequences encoding MT were fused to the autotransporter beta domain of the IgA protease of Neisseria gonorrhoeae, which targeted the hybrid protein toward the bacterial outer membrane. The translocation, surface display, and functionality of the chimeric MTbeta protein was initially demonstrated in Escherichia coli before the transfer of its encoding gene (mtb) to R. eutropha. The resulting bacterial strain, named R. eutropha MTB, was found to have an enhanced ability for immobilizing Cd2+ ions from the external media. Furthermore, the inoculation of Cd2+-polluted soil with R. eutropha MTB decreased significantly the toxic effects of the heavy metal on the growth of tobacco plants (Nicotiana bentamiana). PMID- 10835607 TI - Elevation of the provitamin A content of transgenic tomato plants. AB - Tomato products are the principal dietary sources of lycopene and major source of beta-carotene, both of which have been shown to benefit human health. To enhance the carotenoid content and profile of tomato fruit, we have produced transgenic lines containing a bacterial carotenoid gene (crtI) encoding the enzyme phytoene desaturase, which converts phytoene into lycopene. Expression of this gene in transgenic tomatoes did not elevate total carotenoid levels. However, the beta carotene content increased about threefold, up to 45% of the total carotenoid content. Endogenous carotenoid genes were concurrently upregulated, except for phytoene synthase, which was repressed. The alteration in carotenoid content of these plants did not affect growth and development. Levels of noncarotenoid isoprenoids were unchanged in the transformants. The phenotype has been found to be stable and reproducible over at least four generations. PMID- 10835608 TI - Transgenic avidin maize is resistant to storage insect pests. AB - Avidin is a glycoprotein found in chicken egg white, that sequesters the vitamin biotin. Here we show that when present in maize at levels of > or =100 p.p.m., avidin is toxic to and prevents development of insects that damage grains during storage. Insect toxicity is caused by a biotin deficiency, as shown by prevention of toxicity with biotin supplementation. The avidin maize is not, however, toxic to mice when administered as the sole component of their diet for 21 days. These dates suggest that avidin expression in food or feed grain crops can be used as a biopesticide against a spectrum of stored-produce insect pests. PMID- 10835609 TI - Efficient generation of midbrain and hindbrain neurons from mouse embryonic stem cells. AB - Embryonic stem (ES) cells are clonal cell lines derived from the inner cell mass of the developing blastocyst that can proliferate extensively in vitro and are capable of adopting all the cell fates in a developing embryo. Clinical interest in the use of ES cells has been stimulated by studies showing that isolated human cells with ES properties from the inner cell mass or developing germ cells can provide a source of somatic precursors. Previous studies have defined in vitro conditions for promoting the development of specific somatic fates, specifically, hematopoietic, mesodermal, and neurectodermal. In this study, we present a method for obtaining dopaminergic (DA) and serotonergic neurons in high yield from mouse ES cells in vitro. Furthermore, we demonstrate that the ES cells can be obtained in unlimited numbers and that these neuron types are generated efficiently. We generated CNS progenitor populations from ES cells, expanded these cells and promoted their differentiation into dopaminergic and serotonergic neurons in the presence of mitogen and specific signaling molecules. The differentiation and maturation of neuronal cells was completed after mitogen withdrawal from the growth medium. This experimental system provides a powerful tool for analyzing the molecular mechanisms controlling the functions of these neurons in vitro and in vivo, and potentially for understanding and treating neurodegenerative and psychiatric diseases. PMID- 10835610 TI - Genome-directed primers for selective labeling of bacterial transcripts for DNA microarray analysis. AB - DNA microarrays have the ability to analyze the expression of thousands of the same set of genes under at least two different experimental conditions. However, DNA microarrays require substantial amounts of RNA to generate the probes, especially when bacterial RNA is used for hybridization (50 microg of bacterial total RNA contains approximately 2 microg of mRNA). We have developed a computer based algorithm for prediction of the minimal number of primers to specifically anneal to all genes in a given genome. The algorithm predicts, for example, that 37 oligonucleotides should prime all genes in the Mycobacterium tuberculosis genome. We tested the usefulness of the genome-directed primers (GDPs) in comparison to random primers for gene expression profiling using DNA microarrays. Both types of primers were used to generate fluorescent-labeled probes and to hybridize to an array of 960 mycobacterial genes. Compared to random-primer probes, the GDP probes were more sensitive and more specific, especially when mammalian RNA samples were spiked with mycobacterial RNA. The GDPs were used for gene expression profiling of mycobacterial cultures grown to early log or stationary growth phases. This approach could be useful for accurate genome-wide expression analysis, especially for in vivo gene expression profiling, as well as directed amplification of sequenced genomes. PMID- 10835611 TI - Patents on genes: clarifying the issues. PMID- 10835612 TI - Recent patents in genomics PMID- 10835613 TI - Job compensation in the biotechnology core laboratory. PMID- 10835615 TI - New products PMID- 10835614 TI - People PMID- 10835616 TI - The nature of the number. PMID- 10835617 TI - How to count ... human genes. PMID- 10835618 TI - Better taste through chemistry. PMID- 10835619 TI - A radical solution to death. PMID- 10835620 TI - Panning for pancreatic stem cells. PMID- 10835622 TI - Touching base PMID- 10835621 TI - Lupus, DNase and defective disposal of cellular debris. PMID- 10835623 TI - High-efficiency deleter mice show that FLPe is an alternative to Cre-loxP. PMID- 10835624 TI - Mutations in SPINK5, encoding a serine protease inhibitor, cause Netherton syndrome. AB - We describe here eleven different mutations in SPINK5, encoding the serine protease inhibitor LEKTI, in 13 families with Netherton syndrome (NS, MIM256500). Most of these mutations predict premature termination codons. These results disclose a critical role of SPINK5 in epidermal barrier function and immunity, and suggest a new pathway for high serum IgE levels and atopic manifestations. PMID- 10835625 TI - Mutations of PKD1 in ADPKD2 cysts suggest a pathogenic effect of trans heterozygous mutations. AB - Autosomal dominant polycystic kidney disease (ADPKD) is caused by mutations in PKD1 and PKD2. The products of these genes associate to form heteromeric complexes. Several models have been proposed to explain the mechanism of cyst formation. Here we find somatic mutations of PKD2 in 71% of ADPKD2 cysts analysed. Clonal somatic mutations of PKD1 were identified in a subset of cysts that lacked PKD2 mutations. PMID- 10835626 TI - Quantitative mapping of amplicon structure by array CGH identifies CYP24 as a candidate oncogene. AB - We show here that quantitative measurement of DNA copy number across amplified regions using array comparative genomic hybridization (CGH) may facilitate oncogene identification by providing precise information on the locations of both amplicon boundaries and amplification maxima. Using this analytical capability, we resolved two regions of amplification within an approximately 2-Mb region of recurrent aberration at 20q13.2 in breast cancer. The putative oncogene ZNF217 (ref. 5) mapped to one peak, and CYP24 (encoding vitamin D 24 hydroxylase), whose overexpression is likely to lead to abrogation of growth control mediated by vitamin D, mapped to the other. PMID- 10835627 TI - Computer-based three-dimensional visualization of developmental gene expression. AB - A broad understanding of the relationship between gene activation, pattern formation and morphogenesis will require adequate tools for three-dimensional and, perhaps four-dimensional, representation and analysis of molecular developmental processes. We present a novel, computer-based method for the 3D visualization of embryonic gene expression and morphological structures from serial sections. The information from these automatically aligned 3D reconstructions exceeds that from single-section and whole-mount visualizations of in situ hybridizations. In addition, these 3D models of gene-expression patterns can become a central component of a future developmental database designed for the collection and presentation of digitized, morphological and gene expression data. This work is accompanied by a web site (http://www.univie.ac.at/GeneEMAC). PMID- 10835628 TI - Missense mutations interfere with VEGFR-3 signalling in primary lymphoedema. AB - Primary lymphoedema is a rare, autosomal dominant disorder that leads to a disabling and disfiguring swelling of the extremities and, when untreated, tends to worsen with time. Here we link primary human lymphoedema to the FLT4 locus, encoding vascular endothelial growth factor receptor-3 (VEGFR-3), in several families. All disease-associated alleles analysed had missense mutations and encoded proteins with an inactive tyrosine kinase, preventing downstream gene activation. Our study establishes that VEGFR-3 is important for normal lymphatic vascular function and that mutations interfering with VEGFR-3 signal transduction are a cause of primary lymphoedema. PMID- 10835629 TI - Identification of the familial cylindromatosis tumour-suppressor gene. AB - Familial cylindromatosis is an autosomal dominant genetic predisposition to multiple tumours of the skin appendages. The susceptibility gene (CYLD) has previously been localized to chromosome 16q and has the genetic attributes of a tumour-suppressor gene (recessive oncogene). Here we have identified CYLD by detecting germline mutations in 21 cylindromatosis families and somatic mutations in 1 sporadic and 5 familial cylindromas. All mutations predict truncation or absence of the encoded protein. CYLD encodes three cytoskeletal-associated protein-glycine-conserved (CAP-GLY) domains, which are found in proteins that coordinate the attachment of organelles to microtubules. CYLD also has sequence homology to the catalytic domain of ubiquitin carboxy-terminal hydrolases (UCH). PMID- 10835630 TI - Plzf regulates limb and axial skeletal patterning. AB - The promyelocytic leukaemia zinc finger (Plzf) protein (encoded by the gene Zfp145) belongs to the POZ/zinc-finger family of transcription factors. Here we generate Zfp145-/- mice and show that Plzf is essential for patterning of the limb and axial skeleton. Plzf inactivation results in patterning defects affecting all skeletal structures of the limb, including homeotic transformations of anterior skeletal elements into posterior structures. We demonstrate that Plzf acts as a growth-inhibitory and pro-apoptotic factor in the limb bud. The expression of members of the abdominal b (Abdb) Hox gene complex, as well as genes encoding bone morphogenetic proteins (Bmps), is altered in the developing limb of Zfp145-/- mice. Plzf regulates the expression of these genes in the absence of aberrant polarizing activity and independently of known patterning genes. Zfp145-/- mice also exhibit anterior-directed homeotic transformation throughout the axial skeleton with associated alterations in Hox gene expression. Plzf is therefore a mediator of anterior-to-posterior (AP) patterning in both the axial and appendicular skeleton and acts as a regulator of Hox gene expression. PMID- 10835631 TI - Mutations in RAB27A cause Griscelli syndrome associated with haemophagocytic syndrome. AB - Griscelli syndrome (GS, MIM 214450), a rare, autosomal recessive disorder, results in pigmentary dilution of the skin and the hair, the presence of large clumps of pigment in hair shafts and an accumulation of melanosomes in melanocytes. Most patients also develop an uncontrolled T-lymphocyte and macrophage activation syndrome (known as haemophagocytic syndrome, HS), leading to death in the absence of bone-marrow transplantation. In contrast, early in life some GS patients show a severe neurological impairment without apparent immune abnormalities. We previously mapped the GS locus to chromosome 15q21 and found a mutation in a gene (MYO5A) encoding a molecular motor in two patients. Further linkage analysis suggested a second gene associated with GS was in the same chromosomal region. Homozygosity mapping in additional families narrowed the candidate region to a 3.1-cM interval between D15S1003 and D15S962. We detected mutations in RAB27A, which lies within this interval, in 16 patients with GS. Unlike MYO5A, the GTP-binding protein RAB27A appears to be involved in the control of the immune system, as all patients with RAB27A mutations, but none with the MYO5A mutation, developed HS. In addition, RAB27A-deficient T cells exhibited reduced cytotoxicity and cytolytic granule exocytosis, whereas MYO5A defective T cells did not. RAB27A appears to be a key effector of cytotoxic granule exocytosis, a pathway essential for immune homeostasis. PMID- 10835632 TI - Features of systemic lupus erythematosus in Dnase1-deficient mice. AB - Systemic lupus erythematosus (SLE) is a multifactorial autoimmune disease that affects over one million people in the United States. SLE is characterized by the presence of anti-nuclear antibodies (ANA) directed against naked DNA and entire nucleosomes. It is thought that the resulting immune complexes accumulate in vessel walls, glomeruli and joints and cause a hypersensitivity reaction type III, which manifests as glomerulonephritis, arthritis and general vasculitis. The aetiology of SLE is unknown, but several studies suggest that increased liberation or disturbed clearance of nuclear DNA-protein complexes after cell death may initiate and propagate the disease. Consequently, Dnase1, which is the major nuclease present in serum, urine and secreta, may be responsible for the removal of DNA from nuclear antigens at sites of high cell turnover and thus for the prevention of SLE (refs 7-11). To test this hypothesis, we have generated Dnase1-deficient mice by gene targeting. We report here that these animals show the classical symptoms of SLE, namely the presence of ANA, the deposition of immune complexes in glomeruli and full-blown glomerulonephritis in a Dnase1-dose dependent manner. Moreover, in agreement with earlier reports, we found Dnase1 activities in serum to be lower in SLE patients than in normal subjects. Our findings suggest that lack or reduction of Dnase1 is a critical factor in the initiation of human SLE. PMID- 10835633 TI - Mutations in LHX3 result in a new syndrome revealed by combined pituitary hormone deficiency. AB - Combined pituitary hormone deficiency (CPHD) has been linked with rare abnormalities in genes encoding transcription factors necessary for pituitary development. We have isolated LHX3, a gene involved in a new syndrome, using a candidate-gene approach developed on the basis of documented pituitary abnormalities of a recessive lethal mutation in mice generated by targeted disruption of Lhx3 (ref. 2). LHX3, encoding a member of the LIM class of homeodomain proteins, consists of at least six exons located at 9q34. We identified a homozygous LHX3 defect in patients of two unrelated consanguineous families displaying a complete deficit in all but one (adrenocorticotropin) anterior pituitary hormone and a rigid cervical spine leading to limited head rotation. Two of these patients also displayed a severe pituitary hypoplasia, whereas one patient presented secondarily with an enlarged anterior pituitary. These LHX3 mutations consist of a missense mutation (Y116C) in the LIM2 domain at a phylogenetically conserved residue and an intragenic deletion predicting a severely truncated protein lacking the entire homeodomain. These data are consistent with function of LHX3 in the proper development of all anterior pituitary cell types, except corticotropes, and extrapituitary structures. PMID- 10835634 TI - TLR4 mutations are associated with endotoxin hyporesponsiveness in humans. AB - There is much variability between individuals in the response to inhaled toxins, but it is not known why certain people develop disease when challenged with environmental agents and others remain healthy. To address this, we investigated whether TLR4 (encoding the toll-like receptor-4), which has been shown to affect lipopolysaccharide (LPS) responsiveness in mice, underlies the variability in airway responsiveness to inhaled LPS in humans. Here we show that common, co segregating missense mutations (Asp299Gly and Thr399Ile) affecting the extracellular domain of the TLR4 receptor are associated with a blunted response to inhaled LPS in humans. Transfection of THP-1 cells demonstrates that the Asp299Gly mutation (but not the Thr399Ile mutation) interrupts TLR4-mediated LPS signalling. Moreover, the wild-type allele of TLR4 rescues the LPS hyporesponsive phenotype in either primary airway epithelial cells or alveolar macrophages obtained from individuals with the TLR4 mutations. Our findings provide the first genetic evidence that common mutations in TLR4 are associated with differences in LPS responsiveness in humans, and demonstrate that gene-sequence changes can alter the ability of the host to respond to environmental stress. PMID- 10835635 TI - Homologous recombination is responsible for cell death in the absence of the Sgs1 and Srs2 helicases. AB - DNA helicases are involved in many aspects of DNA metabolism, including transcription, replication, recombination and repair. In the yeast Saccharomyces cerevisiae, the absence of the Sgs1 helicase results in genomic instability and accelerated ageing. In human cells, mutations in orthologues of SGS1 lead to Bloom (BS), Werner (WS) or Rothmund-Thomson (RTS) syndromes, which are rare, autosomal recessive diseases characterized by genetic instability associated with cancer predisposition. Although data concerning these human diseases are accumulating, there is still no clear idea of the function of the proteins involved. Here we show that sgs1Delta mutants are deficient in DNA repair and are defective for induced recombination events that involve homologous chromosomes. The role of homologous recombination is further evidenced in haploid cells in which both Sgs1p and Srs2p are absent. Yeast SRS2 encodes another DNA helicase involved in the maintenance of genome integrity. Our data suggest that some defects observed in BS, WS or RTS are the consequence of unrestrained recombination. PMID- 10835636 TI - Mice deficient for delta- and mu-opioid receptors exhibit opposing alterations of emotional responses. AB - The role of the opioid system in controlling pain, reward and addiction is well established, but its role in regulating other emotional responses is poorly documented in pharmacology. The mu-, delta- and kappa- opioid receptors (encoded by Oprm, Oprd1 and Oprk1, respectively) mediate the biological activity of opioids. We have generated Oprd1-deficient mice and compared the behavioural responses of mice lacking Oprd1, Oprm (ref. 6) and Oprk1 (ref. 7) in several models of anxiety and depression. Our data show no detectable phenotype in Oprk1 /- mutants, suggesting that kappa-receptors do not have a role in this aspect of opioid function; opposing phenotypes in Oprm-/- and Oprd1-/- mutants which contrasts with the classical notion of similar activities of mu- and delta receptors; and consistent anxiogenic- and depressive-like responses in Oprd1-/- mice, indicating that delta-receptor activity contributes to improvement of mood states. We conclude that the Oprd1-encoded receptor, which has been proposed to be a promising target for the clinical management of pain, should also be considered in the treatment of drug addiction and other mood-related disorders. PMID- 10835637 TI - More than 1,000 putative new human signalling proteins revealed by EST data mining. AB - Cloning procedures aided by homology searches of EST databases have accelerated the pace of discovery of new genes, but EST database searching remains an involved and onerous task. More than 1.6 million human EST sequences have been deposited in public databases, making it difficult to identify ESTs that represent new genes. Compounding the problems of scale are difficulties in detection associated with a high sequencing error rate and low sequence similarity between distant homologues. We have developed a new method, coupling BLAST-based searches with a domain identification protocol, that filters candidate homologues. Application of this method in a large-scale analysis of 100 signalling domain families has led to the identification of ESTs representing more than 1,000 novel human signalling genes. The 4,206 publicly available ESTs representing these genes are a valuable resource for rapid cloning of novel human signalling proteins. For example, we were able to identify ESTs of at least 106 new small GTPases, of which 6 are likely to belong to new subfamilies. In some cases, further analyses of genomic DNA led to the discovery of previously unidentified full-length protein sequences. This is exemplified by the in silico cloning (prediction of a gene product sequence using only genomic and EST sequence data) of a new type of GTPase with two catalytic domains. PMID- 10835638 TI - Mutations in TGIF cause holoprosencephaly and link NODAL signalling to human neural axis determination. AB - Holoprosencephaly (HPE) is the most common structural defect of the developing forebrain in humans (1 in 250 conceptuses, 1 in 16,000 live-born infants). HPE is aetiologically heterogeneous, with both environmental and genetic causes. So far, three human HPE genes are known: SHH at chromosome region 7q36 (ref. 6); ZIC2 at 13q32 (ref. 7); and SIX3 at 2p21 (ref. 8). In animal models, genes in the Nodal signalling pathway, such as those mutated in the zebrafish mutants cyclops (refs 9,10), squint (ref. 11) and one-eyed pinhead (oep; ref. 12), cause HPE. Mice heterozygous for null alleles of both Nodal and Smad2 have cyclopia. Here we describe the involvement of the TG-interacting factor (TGIF), a homeodomain protein, in human HPE. We mapped TGIF to the HPE minimal critical region in 18p11.3. Heterozygous mutations in individuals with HPE affect the transcriptional repression domain of TGIF, the DNA-binding domain or the domain that interacts with SMAD2. (The latter is an effector in the signalling pathway of the neural axis developmental factor NODAL, a member of the transforming growth factor-beta (TGF-beta) family.) Several of these mutations cause a loss of TGIF function. Thus, TGIF links the NODAL signalling pathway to the bifurcation of the human forebrain and the establishment of ventral midline structures. PMID- 10835639 TI - Gata3 loss leads to embryonic lethality due to noradrenaline deficiency of the sympathetic nervous system. AB - Mouse embryos deficient in Gata3 die by 11 days post coitum (d.p.c.) from pathology of undetermined origin. We recently showed that Gata3-directed lacZ expression of a 625-kb Gata3 YAC transgene in mice mimics endogenous Gata3 expression, except in thymus and the sympathoadrenal system. As this transgene failed to overcome embryonic lethality (unpublished data and ref. 3) in Gata3-/- mice, we hypothesized that a neuroendocrine deficiency in the sympathetic nervous system (SNS) might cause embryonic lethality in these mutants. We find here that null mutation of Gata3 leads to reduced accumulation of Th (encoding tyrosine hydroxylase, Th) and Dbh (dopamine beta-hydroxylase, Dbh) mRNA, whereas several other SNS genes are unaffected. We show that Th and Dbh deficiencies lead to reduced noradrenaline in the SNS, and that noradrenaline deficiency is a proximal cause of death in mutants by feeding catechol intermediates to pregnant dams, thereby partially averting Gata3 mutation-induced lethality. These older, pharmacologically rescued mutants revealed abnormalities that previously could not be detected in untreated mutants. These late embryonic defects include renal hypoplasia and developmental defects in structures derived from cephalic neural crest cells. Thus we have shown that Gata3 has a role in the differentiation of multiple cell lineages during embryogenesis. PMID- 10835640 TI - Mutations in the gene encoding the serine protease inhibitor, Kazal type 1 are associated with chronic pancreatitis. AB - Chronic pancreatitis (CP) is a continuing or relapsing inflammatory disease of the pancreas. In approximately one-third of all cases, no aetiological factor can be found, and these patients are classified as having idiopathic disease. Pathophysiologically, autodigestion and inflammation may be caused by either increased proteolytic activity or decreased protease inhibition. Several studies have demonstrated mutations in the cationic trypsinogen gene (PRSS1) in patients with hereditary or idiopathic CP. It is thought that these mutations result in increased trypsin activity within the pancreatic parenchyma. Most patients with idiopathic or hereditary CP, however, do not have mutations in PRSS1 (ref. 4). Here we analysed 96 unrelated children and adolescents with CP for mutations in the gene encoding the serine protease inhibitor, Kazal type 1 (SPINK1), a pancreatic trypsin inhibitor. We found mutations in 23% of the patients. In 18 patients, 6 of whom were homozygous, we detected a missense mutation of codon 34 (N34S). We also found four other sequence variants. Our results indicate that mutations in SPINK1 are associated with chronic pancreatitis. PMID- 10835641 TI - Gene transfer by lentiviral vectors is limited by nuclear translocation and rescued by HIV-1 pol sequences. AB - Gene-transfer vectors based on lentiviruses are distinguished by their ability to transduce non-dividing cells. The HIV-1 proteins Matrix, Vpr and Integrase have been implicated in the nuclear import of the viral genome in non-dividing cells. Here we show that a sequence within pol is also required in cis. It contains structural elements previously associated with the progress of reverse transcription in target cells. We restored these elements in cis within late generation lentiviral vectors. The new vector transduced to a much higher efficiency several types of human primary cells, when both growing and growth arrested, including haematopoietic stem cells assayed by long-term repopulation of NOD/SCID mice. On in vivo administration into SCID mice, the vector induced higher plasma levels of human clotting factor IX (F.IX) than non-modified vector. Our results indicate that nuclear translocation of the genome is a rate-limiting step in lentiviral infection of both dividing and non-dividing cells, and that it depends on protein and nucleic acid sequence determinants. Full rescue of this step in lentivirus-based vectors improves performance for gene-therapy applications. PMID- 10835642 TI - Mutations in a gene encoding an ABC transporter cause pseudoxanthoma elasticum. AB - Pseudoxanthoma elasticum (PXE) is a heritable disorder characterized by calcification of elastic fibres in skin, arteries and retina that results in dermal lesions with associated laxity and loss of elasticity, arterial insufficiency and retinal haemorrhages leading to macular degeneration. PXE is usually found as a sporadic disorder, but examples of both autosomal recessive and autosomal dominant forms of PXE have been observed. Partial manifestations of the PXE phenotype have also been described in presumed carriers in PXE families. Linkage of both dominant and recessive forms of PXE to a 5-cM domain on chromosome 16p13.1 has been reported (refs 8,9). We have refined this locus to an 820-kb region containing 6 candidate genes. Here we report the exclusion of five of these genes and the identification of the first mutations responsible for the development of PXE in a gene encoding a protein associated with multidrug resistance (ABCC6). PMID- 10835643 TI - Mutations in ABCC6 cause pseudoxanthoma elasticum. AB - Pseudoxanthoma elasticum (PXE) is a heritable disorder of the connective tissue. PXE patients frequently experience visual field loss and skin lesions, and occasionally cardiovascular complications. Histopathological findings reveal calcification of the elastic fibres and abnormalities of the collagen fibrils. Most PXE patients are sporadic, but autosomal recessive and dominant inheritance are also observed. We previously localized the PXE gene to chromosome 16p13.1 (refs 8,9) and constructed a physical map. Here we describe homozygosity mapping in five PXE families and the detection of deletions or mutations in ABCC6 (formerly MRP6) associated with all genetic forms of PXE in seven patients or families. PMID- 10835644 TI - Analysis of expressed sequence tags indicates 35,000 human genes. AB - The number of protein-coding genes in an organism provides a useful first measure of its molecular complexity. Single-celled prokaryotes and eukaryotes typically have a few thousand genes; for example, Escherichia coli has 4,300 and Saccharomyces cerevisiae has 6,000. Evolution of multicellularity appears to have been accompanied by a several-fold increase in gene number, the invertebrates Caenorhabditis elegans and Drosophila melanogaster having 19,000 and 13,600 genes, respectively. Here we estimate the number of human genes by comparing a set of human expressed sequence tag (EST) contigs with human chromosome 22 and with a non-redundant set of mRNA sequences. The two comparisons give mutually consistent estimates of approximately 35,000 genes, substantially lower than most previous estimates. Evolution of the increased physiological complexity of vertebrates may therefore have depended more on the combinatorial diversification of regulatory networks or alternative splicing than on a substantial increase in gene number. PMID- 10835645 TI - Estimate of human gene number provided by genome-wide analysis using Tetraodon nigroviridis DNA sequence. AB - The number of genes in the human genome is unknown, with estimates ranging from 50,000 to 90,000 (refs 1, 2), and to more than 140,000 according to unpublished sources. We have developed 'Exofish', a procedure based on homology searches, to identify human genes quickly and reliably. This method relies on the sequence of another vertebrate, the pufferfish Tetraodon nigroviridis, to detect conserved sequences with a very low background. Similar to Fugu rubripes, a marine pufferfish proposed by Brenner et al. as a model for genomic studies, T. nigroviridis is a more practical alternative with a genome also eight times more compact than that of human. Many comparisons have been made between F. rubripes and human DNA that demonstrate the potential of comparative genomics using the pufferfish genome. Application of Exofish to the December version of the working draft sequence of the human genome and to Unigene showed that the human genome contains 28,000-34,000 genes, and that Unigene contains less than 40% of the protein-coding fraction of the human genome. PMID- 10835647 TI - De novo deletions of SNRPN exon 1 in early human and mouse embryos result in a paternal to maternal imprint switch PMID- 10835646 TI - Gene index analysis of the human genome estimates approximately 120,000 genes. AB - Although sequencing of the human genome will soon be completed, gene identification and annotation remains a challenge. Early estimates suggested that there might be 60,000-100,000 (ref. 1) human genes, but recent analyses of the available data from EST sequencing projects have estimated as few as 45,000 (ref. 2) or as many as 140, 000 (ref. 3) distinct genes. The Chromosome 22 Sequencing Consortium estimated a minimum of 45,000 genes based on their annotation of the complete chromosome, although their data suggests there may be additional genes. The nearly 2,000,000 human ESTs in dbEST provide an important resource for gene identification and genome annotation, but these single-pass sequences must be carefully analysed to remove contaminating sequences, including those from genomic DNA, spurious transcription, and vector and bacterial sequences. We have developed a highly refined and rigorously tested protocol for cleaning, clustering and assembling EST sequences to produce high-fidelity consensus sequences for the represented genes (F.L. et al., manuscript submitted) and used this to create the TIGR Gene Indices-databases of expressed genes for human, mouse, rat and other species (http://www.tigr.org/tdb/tgi.html). Using highly refined and tested algorithms for EST analysis, we have arrived at two independent estimates indicating the human genome contains approximately 120,000 genes. PMID- 10835648 TI - Physician-scientists: mind the gap. PMID- 10835649 TI - Peripheral vasoconstriction during REM sleep detected by a new plethysmographic method. PMID- 10835650 TI - In vitro models of mucosal HIV transmission. PMID- 10835651 TI - Reply to 'In vitro models of mucosal HIV transmission' PMID- 10835652 TI - No dismissal for hate-mail author. PMID- 10835653 TI - Experiment reveals deficits in european regulation PMID- 10835654 TI - Can veronesi transform italian research? PMID- 10835655 TI - NIH researchers receive cut-price BRCA test. PMID- 10835656 TI - Transplant organ allocation squabble continues PMID- 10835657 TI - HHS criticizes research centers for inaction on patient safety. PMID- 10835658 TI - The NIH improves its record on women in trials. PMID- 10835659 TI - New economic analysis draws big money to malaria. PMID- 10835660 TI - Australia to create biomedical lobby group PMID- 10835661 TI - Baylor faculty upset over science and religion center. PMID- 10835662 TI - World AIDS conference PMID- 10835664 TI - Researcher stockpiled salmonella and explosives PMID- 10835663 TI - Congress debates stem cell research. PMID- 10835665 TI - The Declaration of Helsinki, CIOMS and the ethics of research on vulnerable populations. PMID- 10835667 TI - Darwin's ghost: the origin of species updated PMID- 10835666 TI - Public-private partnerships for public health. PMID- 10835668 TI - Neural transplantation PMID- 10835669 TI - Gene therapy for human SCID: dreams become reality. PMID- 10835670 TI - Gene therapy turns the corner. PMID- 10835671 TI - HIV nuclear import: What's the flap? PMID- 10835672 TI - Prostate cancer schemes for androgen escape. PMID- 10835673 TI - A dual function for CD40 agonists. PMID- 10835674 TI - Viral damage or 'molecular mimicry'-placing the blame in myocarditis. PMID- 10835675 TI - Tying-up tuberculosis. PMID- 10835676 TI - Amyloids: tombstones or triggers? PMID- 10835677 TI - Is neural cell transplantation ready for the clinic? PMID- 10835678 TI - Research news PMID- 10835679 TI - The 3' untranslated region of messenger RNA: A molecular 'hotspot' for pathology? AB - The role of the 3' untranslated region in posttranscriptional regulation of mRNA expression is being elucidated. Here we describe diseases arising from anomalies in this region, that affect the expression of one or more genes. PMID- 10835680 TI - Receptor-dependent cell stress and amyloid accumulation in systemic amyloidosis. AB - Accumulation of fibrils composed of amyloid A in tissues resulting in displacement of normal structures and cellular dysfunction is the characteristic feature of systemic amyloidoses. Here we show that RAGE, a multiligand immunoglobulin superfamily cell surface molecule, is a receptor for the amyloidogenic form of serum amyloid A. Interactions between RAGE and amyloid A induced cellular perturbation. In a mouse model, amyloid A accumulation, evidence of cell stress and expression of RAGE were closely linked. Antagonizing RAGE suppressed cell stress and amyloid deposition in mouse spleens. These data indicate that RAGE is a potential target for inhibiting accumulation of amyloid A and for limiting cellular dysfunction induced by amyloid A. PMID- 10835681 TI - Efficient retrovirus-mediated transfer of the multidrug resistance 1 gene into autologous human long-term repopulating hematopoietic stem cells. AB - Pre-clinical studies indicate that efficient retrovirus-mediated gene transfer into hematopoietic stem cells and progenitor cells can be achieved by co localizing retroviral particles and target cells on specific adhesion domains of fibronectin. In this pilot study, we used this technique to transfer the human multidrug resistance 1 gene into stem and progenitor cells of patients with germ cell tumors undergoing autologous transplantation. There was efficient gene transfer into stem and progenitor cells in the presence of recombinant fibronectin fragment CH-296. The infusion of these cells was associated with no harmful effects and led to prompt hematopoietic recovery. There was in vivo vector expression, but it may have been limited by the high rate of aberrant splicing of the multidrug resistance 1 gene in the vector. Gene marking has persisted more than a year at levels higher than previously reported in humans. PMID- 10835683 TI - Immunoglobulin framework-derived peptides function as cytotoxic T-cell epitopes commonly expressed in B-cell malignancies. AB - Although the idiotypic structures of immunoglobulin from malignant B cells were the first tumor-specific determinants recognized, and clinical vaccination trials have demonstrated induction of tumor-specific immunity, the function of immunoglobulin-specific CD8+ cytotoxic T lymphocytes in tumor rejection remains elusive. Here, we combined bioinformatics and a T cell-expansion system to identify human immunoglobulin-derived peptides capable of inducing cytotoxic T lymphocyte responses. Immunogenic peptides were derived from framework regions of the variable regions of the immunoglobulin that were shared among patients. Human leukocyte-antigen-matched and autologous cytotoxic T lymphocytes specific for these peptides killed primary malignant B cells, demonstrating that malignant B cells are capable of processing and presenting such peptides. Targeting shared peptides to induce T-cell responses might further improve current vaccination strategies in B-cell malignancies. PMID- 10835682 TI - Limitin: An interferon-like cytokine that preferentially influences B-lymphocyte precursors. AB - We have identified an interferon-like cytokine, limitin, on the basis of its ability to arrest the growth of or kill lympho-hematopoietic cells. Limitin strongly inhibited B lymphopoiesis in vitro and in vivo but had little influence on either myelopoiesis or erythropoiesis. Because limitin uses the interferon alpha/beta receptors and induces interferon regulatory factor-1, it may represent a previously unknown type I interferon prototype. However, preferential B-lineage growth inhibition and activation of Janus kinase 2 in a myelomonocytic leukemia line have not been described for previously known interferons. PMID- 10835684 TI - Therapeutic activity of agonistic monoclonal antibodies against CD40 in a chronic autoimmune inflammatory process. AB - The use of agonistic monoclonal antibody against CD40 has emerged as one the most effective ways to boost immune responses against infectious agents or to fight cancer. Here, we report that the same monoclonal antibodies against CD40 (FGK45 and 3/23) previously used to elicit protective immune responses treated the autoimmune inflammatory process of chronic collagen-induced arthritis in DBA/1 TCR-beta transgenic mice, as well as collagen-induced arthritis in DBA/1 mice, both animal models of rheumatoid arthritis. This study indicates that agonistic monoclonal antibody against CD40 can potentially be used to treat chronic autoimmune inflammatory processes. PMID- 10835685 TI - FcalphaRI-positive liver Kupffer cells: reappraisal of the function of immunoglobulin A in immunity. AB - Despite the well-recognized involvement of immunoglobulin (Ig) A in mucosal immunity, the function of its receptor, FcalphaRI (CD89), is poorly understood. The ability of FcalphaRI to activate leukocytes seems to conflict with the proposed anti-inflammatory activity of secretory IgA. We show here that in a transgenic mouse model, inflammatory mediators induced expression of FcalphaRI on Kupffer cells, which enabled efficient phagocytosis in vivo of bacteria coated with serum IgA. Secretory IgA did not initiate phagocytosis. Therefore, interactions between serum IgA and FcalphaRI on Kupffer cells may provide a 'second line of defense' in mucosal immunity, by eliminating invasive bacteria entering through the portal circulation and thus preventing disease. PMID- 10835686 TI - Immunologic 'ignorance' of vascularized organ transplants in the absence of secondary lymphoid tissue. AB - Secondary lymphoid organs (the spleen, lymph nodes and mucosal lymphoid tissues) provide the proper environment for antigen-presenting cells to interact with and activate naive T and B lymphocytes. Although it is generally accepted that secondary lymphoid organs are essential for initiating immune responses to microbial antigens and to skin allografts, the prevailing view has been that the immune response to primarily vascularized organ transplants such as hearts and kidneys does not require the presence of secondary lymphoid tissue. The assumption has been that the immune response to such organs is initiated in the graft itself when recipient lymphocytes encounter foreign histocompatibility antigens presented by the graft's endothelial cells. In contrast to this view, we show here that cardiac allografts are accepted indefinitely in recipient mice that lack secondary lymphoid tissue, indicating that the alloimmune response to a vascularized organ transplant cannot be initiated in the graft itself. Moreover, we demonstrate that the permanent acceptance of these grafts is not due to tolerance but is because of immunologic 'ignorance'. PMID- 10835687 TI - A principal target of human immunity to malaria identified by molecular population genetic and immunological analyses. AB - New strategies are required to identify the most important targets of protective immunity in complex eukaryotic pathogens. Natural selection maintains allelic variation in some antigens of the malaria parasite Plasmodium falciparum. Analysis of allele frequency distributions could identify the loci under most intense selection. The merozoite surface protein 1 (Msp1) is the most-abundant surface component on the erythrocyte-invading stage of P. falciparum. Immunization with whole Msp1 has protected monkeys completely against homologous and partially against non-homologous parasite strains. The single-copy msp1 gene, of about 5 kilobases, has highly divergent alleles with stable frequencies in endemic populations. To identify the region of msp1 under strongest selection to maintain alleles within populations, we studied multiple intragenic sequence loci in populations in different regions of Africa and Southeast Asia. On both continents, the locus with the lowest inter-population variance in allele frequencies was block 2, indicating selection in this part of the gene. To test the hypothesis of immune selection, we undertook a large prospective longitudinal cohort study. This demonstrated that serum IgG antibodies against each of the two most frequent allelic types of block 2 of the protein were strongly associated with protection from P. falciparum malaria. PMID- 10835688 TI - Pancreatic expression of interferon-gamma protects mice from lethal coxsackievirus B3 infection and subsequent myocarditis. AB - Cardiovascular disease is one of the leading causes of death worldwide, and has been associated with many environmental risk factors. Recent evidence has indicated the involvement of pathogens such as viruses as causative agents, and specifically identified the coxsackievirus B serogroup as the leading culprit. Not only has coxsackievirus B3 (CB3) been identified from patients with cardiovascular disease, but also infection of mice with CB3 strains can reproduce human clinical heart disease in rodents. Several mechanisms have been proposed in an attempt to distinguish between pathology mediated by direct viral destruction of cardiac muscle cells or by the virus-induced immune response directed at infected myocytes or at 'mimicked' epitopes shared between viral and cardiac antigens. To distinguish between these mechanisms, we infected a unique mouse that diminishes the extent of infection and spread of the virus, but allows complete immunity to the virus. Transgenic mice expressing interferon-gamma in their pancreatic beta cells failed to develop CB-3-induced myocarditis. This work challenges the idea of the function of the immune response and 'molecular mimicry' in the CB-3-induced autoimmune myocarditis model, and instead favors the idea of virus-mediated damage. These results emphasize the benefit of reducing the level of viremia early during infection, thereby reducing the incidence of virus-mediated heart damage and autoimmunity. PMID- 10835689 TI - Complete reversal of fatal pulmonary hypertension in rats by a serine elastase inhibitor. AB - Progression of pulmonary hypertension is associated with increased serine elastase activity and the proteinase-dependent deposition of the extracellular matrix smooth muscle cell survival factor tenascin-C (refs. 1,2). Tenascin-C amplifies the response of smooth muscle cells to growth factors, which are also liberated through matrix proteolysis. Recent organ culture studies using hypertrophied rat pulmonary arteries have shown that elastase inhibitors suppress tenascin-C and induce smooth muscle cell apoptosis. This initiates complete regression of the hypertrophied vessel wall by a coordinated loss of cellularity and extracellular matrix. We now report that elastase inhibitors can reverse advanced pulmonary vascular disease produced in rats by injecting monocrotaline, an endothelial toxin. We began oral administration of the peptidyl trifluoromethylketone serine elastase inhibitors M249314 or ZD0892 21 days after injection of monocrotaline. A 1-week treatment resulted in 92% survival, compared with 39% survival in untreated or vehicle-treated rats. Pulmonary artery pressure and muscularization were reduced by myocyte apoptosis and loss of extracellular matrix, specifically elastin and tenascin-C. After 2 weeks, pulmonary artery pressure and structure normalized, and survival was 86%, compared with 0% in untreated or vehicle-treated rats. Although concomitant treatment with various agents can reduce pulmonary hypertension, we have documented complete regression after establishment of malignant monocrotaline-induced disease. PMID- 10835690 TI - Glucocorticoids can promote androgen-independent growth of prostate cancer cells through a mutated androgen receptor. AB - The androgen receptor (AR) is involved in the development, growth and progression of prostate cancer (CaP). CaP often progresses from an androgen-dependent to an androgen-independent tumor, making androgen ablation therapy ineffective. However, the mechanisms for the development of androgen-independent CaP are unclear. More than 80% of clinically androgen-independent prostate tumors show high levels of AR expression. In some CaPs, AR levels are increased because of gene amplification and/or overexpression, whereas in others, the AR is mutated. Nonetheless, the involvement of the AR in the transition of CaP to androgen independent growth and the subsequent failure of endocrine therapy are not fully understood. Here we show that in CaP cells from a patient who failed androgen ablation therapy, a doubly mutated AR functioned as a high-affinity cortisol/cortisone receptor (ARccr). Cortisol, the main circulating glucocorticoid, and its metabolite, cortisone, both equally stimulate the growth of these CaP cells and increase the secretion of prostate-specific antigen in the absence of androgens. The physiological concentrations of free cortisol and total cortisone in men greatly exceed the binding affinity of the ARccr and would activate the receptor, promoting CaP cell proliferation. Our data demonstrate a previously unknown mechanism for the androgen-independent growth of advanced CaP. Understanding this mechanism and recognizing the presence of glucocorticoid responsive AR mutants are important for the development of new forms of therapy for the treatment of this subset of CaP. PMID- 10835691 TI - Efficient detection and immunomagnetic sorting of specific T cells using multimers of MHC class I and peptide with reduced CD8 binding. PMID- 10835692 TI - Assessing tumors in living animals through measurement of urinary beta-human chorionic gonadotropin. PMID- 10835693 TI - On the market PMID- 10835694 TI - [Past and present of Medicina (Buenos Aires)]. AB - To celebrate the 60th anniversary of Medicina (Buenos Aires) an International Symposium was held at the National Academy of Medicine of Buenos Aires on the 6 7th of October 1999, under the title of Clinical investigation in the next millennium. This meeting was a success as evidenced by the 376 registered attendants. Sixty years of uninterrupted publication is an uncommon feat in our midst and this could be achieved on the basis of a number of factors which include, the initiative of those who founded the journal, the unfailing motivation and dedication of the Editorial Board and primarily the authors who have trusted us with their manuscripts. Of the many important papers published, we have selected a few which proved to be milestones in the development of Argentine biomedicine. It is to be hoped that the future will bring an increase in our impact index through more and even better papers eventually reflecting the authentic scientific value of our country. PMID- 10835695 TI - [Molecular biology and medicine at the end of the 20th Century]. AB - This paper reviews basic concepts of modern molecular biology with the premise that its influence in today's medicine is so important that its knowledge cannot remain limited to a few experts. I first analyze the overall structure and organization of human genes, their split nature and the flow of genetic information from DNA to protein. The role of transcriptional control in the regulation of gene expression and cell differentiation is described by introducing experimental examples that define the importance of "master" genes. Basic concepts of genetic engineering, the generation of transgenic and knock out animals and the uses of molecular biology in clinical diagnosis, paternity tests and forensic medicine are presented. Finally, I discuss the possibilities of gene therapy and the fantasies and realities of transgenesis and cloning by nuclear transplant in humans. PMID- 10835696 TI - [Tuberculosis focused through the molecular biology zoom]. AB - The sequence of Mycobacterium tuberculosis genome set up a process of systematic research and improved the understanding of the microorganism biology, albeit the clues of its virulence and pathogenicity still remain rather cryptic. Genetic engineering did not succeed in designing more effective or convenient tools to accomplish the control of tuberculosis. Neither the bacillus variants created by mutagenesis and recombination nor the microorganism subunits (antigens, DNA) proved to be significantly better than the BCG vaccine as immunogens. Likewise, the search for novel bacterial targets and more active antibiotics has been unfruitful thus far, even though some advance in drugs formula or delivery systems is in progress. Conversely, new and ingenious instruments have been developed to optimize the epidemiological tracing and diagnosis of the disease. The finding of strain molecular markers consolidated the investigation of tuberculosis spread and revealed the expansion and prevalence of certain lineages of the bacillus in some ecological niches. Molecular methods are specific to immediately characterize the bacilli detected by microscopy or culture which resulted in rapid diagnosis build-up. This improvement is decisive for immunodepressed patients and those affected by multidrug-resistant tuberculosis. To be meaningful and precise the results produced by molecular investigations must be properly backed up by conventional microbiology, pathology and epidemiology. PMID- 10835697 TI - [Applications of gene therapy in medical genetics]. AB - Over the past three decades, an increasing proportion of genetic research has consisted of molecular studies in medicine. It has resulted in a profound change in the understanding of the pathophysiology of diverse genetic diseases. Gene therapy is the use of nucleic acids as therapeutically useful molecules. Although many genetic discoveries have resulted in better diagnostic tests, the application of molecular technologies to the treatment of genetic diseases is natural and logical. Gene therapy is in a phase of its youth, nevertheless it holds very real promise. In the first 9 years, 396 clinical protocols have been approved worldwide and over 3,000 patients from 22 different countries have carried genetically engineered cells in their body. The conclusion from these trials are that gene therapy has the potential for treating a broad array of human diseases and the procedure appears to carry a very low risk of adverse reactions, but the efficiency of gene transfer and expression in human patients is low. No formal phase III studies to establish clinical efficacy have been completed. Gene therapy is potentially a powerful clinical approach, but it has been restricted by the limited knowledge of vectors and pathophysiology of the diseases to be treated. Better understanding of the disease processes, improvements in vector design, and a great attention to the pharmacological aspects should permit the development of more effective gene therapy. PMID- 10835698 TI - [Immunology of the future]. AB - Immunology is a young science, much younger than MEDICINA, since its modern biological foundations were established only in the 70s-80s with the definition of the T and B cell receptors. In spite of its relative youth, many key basic biological knowledge such as signal transduction, gene recombination, apoptosis and cell cycle regulation have been initiated or expanded using cells from the immune system. Although it is not easy to predict the future, we include in this essay eight areas in which we can expect important developments. These include the definition of the biological basis of homeostasis, the exact mechanisms of antigen processing and presentation by professional antigen presenting cells (which implies also the redefinition of our notions of antigen, now based on the recognition of small peptides from larger molecules), the analysis of multifactorial interactions, the sociology of cell interactions based on adhesion molecules and components of the extracellular matrix, the development of useful antagonists of active inflammatory molecules, the definition of the properties of regional immune systems (such as skin and gut), the definition of the immunities against parasites and finally the definition of the structural basis for autoimmunity. The advances in these areas (and in others which are now impossible to predict) will produce, we think, immunological therapies for prevention (vaccines) and treatment (antagonists, etc.) of infectious, parasitic and other forms of inflammatory diseases, including autoimmunity. PMID- 10835699 TI - [The renin saga]. AB - This saga is the story of a scientific development. From the search of a mechanism to explain high blood pressure, research was orientated to the functions of an omnipresent biochemical system. And from the search to elucidate the etiology of arterial hypertension, research has ended up studying the local, functional and structural activity of the renin-angiotensin system and the possibilities of interfering with its actions. Since Bright, left ventricular hypertrophy became associated with nephrosclerosis. Later on, clinical studies led Volhard and Fahr to associate nephrosclerosis to high blood pressure while biochemical research led Tigerstedt and Bergmann to demonstrate that renin was associated to high blood pressure. Two teams of investigators, one in Argentina and one in USA discovered the biochemical mechanism by which renin acted on arterioles and later on, two other teams, one in USA and one in England, discovered the biochemical steps leading to the synthesis of angiotensin II. Since Goldblatt's experimental design resulting in a reliable method to obtain arterial hypertension, more than 20 years had to elapse before renal artery stenosis became established as the main cause of clinical secondary arterial hypertension. The renin-angiotensin system became part of a very complex array of substances able to regulate local circulation directly or indirectly and angiotensin has become involved in the remodeling of the smooth muscles of arterioles and myocardium. PMID- 10835700 TI - [Thrombotic microangiopathies: HUS/TTP. Physiopathological aspects]. AB - In thrombotic thrombocytopenic purpura (TTP) and in the hemolytic uremic syndrome (HUS) fibrin-platelet thrombi occlude arterioles and capillaries. The mechanism of endothelial cell injury and the mechanism of thrombosis are the most important physiopathological events in this pathology and are largely unknown. In HUS due to the Shiga toxin, the lesion of the endothelial cells is due to penetration of the toxin into the cell via the Gb3 receptor. Endothelial cell death is a consequence of altered protein synthesis at the ribosomal level. Cytokines released during the inflammatory process, possibly enhance the endothelial damage. Genetic and immunologic predisposing factors for the development of HUS have also been postulated. In idiopathic, secondary and familial HUS/TTP the mechanism of endothelial lesion is unknown, but multiple responsible factors have been advocated such as infections, drugs, pregnancy, autoantibodies, apoptosis inducing molecules, etc. and other genetic, hormonal or immunologic predisposing factors may also be involved. Factor H deficiency has been blamed in familiar cases. The most important cause of microcirculation thrombosis is the thrombogenic capacity of endothelial cell "activation" or injury induced by multiple mechanisms. The predominant source of plasma vW factor multimers is apparent in the altered endothelial cell. The unusually large vWF multimers are more effective at binding to platelet glycoprotein Ib-IX and IIb-IIIa complexes and inducing aggregation, as also occurred with the low weight multimers formed with excessive proteolysis, as described in the acute phase of HUS/TTP. The recent report of congenital deficiency of a vWF protease in familial TTP and its functional inhibition by autoantibodies in acquired cases is characteristic of TTP. This protease inhibition has never been described in HUS and might represent pathogenetic differences between TTP and HUS, and contribute to the differential diagnosis, but further confirmation of these findings is needed. We postulate that the abnormal cleavage of the vWF subunit, with formation of different multimers with increased platelet aggregating capacity is an important mechanism to increase the microcirculatory thrombosis, but it is only a partial aspect in a more complex and unknown thrombogenic stimulation secondary to the endothelial lesion or activation. Better knowledge of the endothelial physiology and the genetic polymorphism of the endothelial cell, the clonation of vWF-cleavage protease, etc., will provide valuable tools for the understanding of these fascinating entities. PMID- 10835701 TI - [Molecular genetics of essential hypertension. Susceptibility and resistance genes]. AB - Essential Hypertension (EH) is a multifactorial and polygenic syndrome with a high impact in public health. Recently, rare mendelian forms of hypertension such as glucocorticoid-remediable aldosteronism (GRA), apparent mineralocorticoid excess (AME) and Liddle Syndrome caused by single gene mutations have been identified in which the mechanism is an increased sodium retention. Therefore, it is tempting to speculate that the most common forms of EH may be due to diverse highly prevalent molecular variants of susceptibility genes with low penetrance that are involved in arterial blood pressure (ABP) and electrolytic balance. Although a number of candidate genes such as NO synthases, ANP, ion transporters, adducins, LDL receptor, etc. can participate, renin-angiotensin system components are the most extensively studied. Although not associated with EH, the ACE D allele seems to confer a high risk of CHD or LVH. Angiotensinogen 235T and 174M variants are more likely associated with EH and positively correlate with clinical or ambulatory ABP in adolescents or adults. Individuals who carry these angiotensinogen alleles would be at 1.4 higher risk of suffering EH than homozygotes for M235 or T174 alleles. Associations of AT1 receptor variants with EH remain to be definitively defined. In conclusion, the characterization of the genetic background, although difficult at the present time, may have clear benefits in terms of defining a more rational therapy and prevention in individuals at risk. Even though this aim seems difficult to achieve since more than 150 candidate genes have been postulated as the cause of EH, with 6 to 10 SNPs in each of them, new technologies such as DNA micro-arrays will provide us with the opportunity to analyse the total genetic risk in each subject. PMID- 10835702 TI - [Present and future of immunosuppressive therapy]. AB - Immunosuppressive drugs make possible the acceptance of organ allografts among individuals with differences in Major Histocompatibility Antigens (HLA). Transplantation of vital organs prolongs the survival of patients with terminal diseases, and this procedure has become a routine practice in the clinic, mainly because of advances in immunosuppressive therapy. Some immunosuppressive drugs, such as glucocorticosteroids and azathioprine, have been used for the past 30 years. More recently, newly discovered agents with a better ratio of efficacy to toxicity have been added to the armamentarium of anti-rejection therapies. Progress in understanding T cell activation in response to alloantigens has contributed to the development of new and more selective strategies to control the immune response and prevent acute rejection. The use of drugs in combination, with or without monoclonal antibodies, has also improved the efficacy and reduced the toxicity of immunosuppressive therapies. The new agents include drugs that interfere with calcineurin, inhibitors of de novo purine biosynthesis, kinase inhibitors, as well as monoclonal antibodies that block activation signals on the surface of T cells or co-stimulatory signals between T cells and antigen presenting cells. In this review the modes of action of commonly used immunosuppressive drugs are described. Successful new strategies are also being developed to establish tolerance to allografts in rodents and non-human primates. The progress in these approaches, although still in the experimental stages, offers promising alternatives for these patients in the future. Treatment protocols using combinations of drugs with antibodies that might produce tolerance in humans are also discussed. PMID- 10835703 TI - [Medical ethics in the world's market. Hippocratic fidelity or enterprise fidelity]. AB - The advance which resulted in the mean survival increase from 50 to 75 years between 1920 and 1990 also provoked the rise in health care costs, and the so called "health crisis". In order to contain it, market tactics were put to action, health care was considered a commodity, patients "consumers" and hospitals or physicians "providers". Economists, accountants and business advisors in charge of "Health Maintenance Organizations" (HMO) started the very profitable activity of intervening between patients and physicians. Rationing, use of general practice guides, suboptimal treatments, risk avoidance and other market tactics changed the practice of a profession into a business enterprise. The HMO decides if, when, how and how much will be given to any "consumer". Use of technology more impersonal and easily administered is the leading feature of to-day's medicine over the intellectual activity of the physician who hears, understands, makes the physical examination, diagnosis and treatment. The increasing depreciation of his task obliges the physician to enlarge the number and decrease his communication with his patients. His fiduciary obligation is subordinated to market needs and his practice increasingly compromises his moral integrity. The HMO boasts of the quality of the service given, this is the timely use of to-day's appropriate resources. Nobody wants the 1950 car or medical practice. Tomorrow's practice however depends on increasing knowledge, that is, on research, an activity which is not the HMO object. The academic-medical center, the very place where the interaction of teaching and investigating promotes the excellence is discriminated by the HMO because of its compromise with fiduciary activity imposed by 2500 years of jewish-christian philosophy. The future of these institutions (state's Cinderella's) is progressively compromised; when we loose them how long will it take to recover them? The politicians are always ready to create new hospitals, after they are built consuming large amounts of money, they become disinterested. All hospitals in our country are completely active only 4 hours/day, their physicians travel afterwards to their diverse places of activity consuming much of their time in getting there and complying with the bureaucratic tasks imposed by HMO. In our country with 14% unemployment and 1/3 of the population without any health coverage, the institution of universal health insurance is mandatory. Preventive medicine is not effective for people who lack the means for adequate nutrition, education or transportation, they do not visit doctors or use medicines. PMID- 10835704 TI - [Scientific research in France. Structures and research career]. AB - In France, scientific research has always been considered an important activity. The aim of this article is to present data with reference to French research, mainly research in Medical and Biological Sciences. This includes a description of the Research Career in the two main institutions, CNRS (National Research Center) and INSERM (National Institute of Health and Medical Research). Investment in science is substantial in France originating both from the Scientific Institutions of the State and from a large number of Private Organizations. France is the second European country, after Germany, as far as the number of national and private investigators is concerned. CNRS covers all the dominions of science, including Natural Sciences, Mathematics, Biology and Medical Sciences. One of its main structures is undoubtedly its National Committee of Research which is responsible for the evaluation of the laboratories and research projects, of the incorporation of investigators in the Research Career and of their promotion. This Committee includes, among others, members of the Research Career elected among themselves. In parallel, INSERM is centered in both basic and applied Medical Research, and is supervised by a Scientific Committee and eleven specialized Commissions. In both institutions, the Research Career incorporates investigators as civil servants after very competitive examinations; their performance is evaluated by specialized committees as well as by national and international experts. PMID- 10835705 TI - [Research career in Argentina]. AB - The National Research Council of Argentina (CONICET) was founded in 1958, and the Research Career opened officially two years later (1960). Originally, 214 scientists belonged to this Career, increasing slowly to 3642 members in 1999. There are 5 categories of investigators, besides the Clinical Investigator class for the area of Medical Sciences. Investigators comprise 46%, while Technicians (34%) and Fellows (20%) account for the rest of CONICET research personnel. The low number of Fellows is a matter of worry, although Fellows from universities, local councils and private foundations contribute to increase their total number. There is an irregular regional distribution of Investigators, most of whom work in the Federal Capital and Province of Buenos Aires (61%). Increasing the salary of those living outside the metropolitan area did not solve the problem. Input to the Research Career has been erratic and not well planned, while mechanisms for personnel output due to low productivity or retirement age has had variable and erratic policies. The problem the Research Career is facing is similar to that of other areas of CONICET, due to an extremely low budget. Hopefully, new CONICET authorities will be active scientists considering Science and Technology as a Matter of State, just as important for the country as health, education or recovery of the Malvinas Islands. PMID- 10835706 TI - [Competitiveness in science. Today, tomorrow, and forever]. AB - Paternalistic governments and highly bureaucratized administrations produce mediocre science policy decisions that often allow for the co-existence of potentially competitive scientists alongside with those that are not. This invariably results in failure to produce significant research. It seems apparent therefore, that policy change aiming at improving science and technology must begin with intensification of the level of individual competitiveness. Nations that have internationally competitive levels of technical and scientific activity such as Japan, USA and Canada, share in common certain features that foster individual competitiveness despite the fact that their socioeconomic basis are vastly different. These common features include administrative continuity, very high academic standards and a highly educated work force. The scientist's emotional cost in competitive environments is high but there seems to be no alternative given the sophistication of the topics that are dealt with in formerly purely descriptive sciences such as biomedicine, and given the enormous speed of electronic communications. The role of governments in fostering science and technology should be mainly concerned with conducting a sound fiscal policy in order to provide for the needs of education and scientific activity. Governments can also play a key role in insuring that science remains competitive through the delineation of rules that increase individual competitiveness rather than with policy schemes that fail to directly address the responsibility of the individual. Policies to increase individuals' performance may prove costly to politicians given that these adjustments imply unpopular decisions regarding an increase in academic performance expectation beginning in high school and the re assignment of functions of individuals or institutions that do not meet international productivity criteria. PMID- 10835707 TI - Diet and nutrition of prehistoric populations at the alluvial banks of the Parana River. AB - This study attempts to characterize the health status and diet of prehistoric populations (1,000-2,000 years BP), dwelling at both banks of Parana River, between 29 degrees S and 32 degrees S. The data obtained suggest that these prehistoric populations had an adequate nutritional status, with complete proteins in the diet, as suggested by the ratio strontium/calcium in their bone mineral (0.71 +/- 0.04 microgram Srx1,000/mg Ca). The overall frequency of dental caries (4.9%) coincides with that reported for hunters-gatherers. The average mineral densities of the tibiae of adult subjects exhumed at two sites (males: 1.51 +/- 0.07 gr/cm2; females: 1.24 +/- 0.06 gr/cm2) suggested that they had significant bone mass, an asset compatible with adequate nutrition. In metacarpals, the amount of cortical tissue also suggests bone mass comparable to contemporaneous controls. The growth and development of the prehistoric populations studied are deemed normal as shown by the clear sexual dimorphism of their estimated heights at adult age (males: 177-183 cm; females 152-166 cm) and their bone mass. PMID- 10835708 TI - [Eight years of experience in a single institution in hematopoietic stem cell autologous transplantation in malignant hematological diseases and in solid tumors]. AB - Between August 1991 and December 1998, 400 patients (lymphomas: 197; acute leukemia: 86; multiple myeloma: 70 and solid tumors: 47) were admitted for autologous transplantation. All patients were mobilized with chemotherapy plus G CSF. The hematological recovery was similar in all disease groups. Patients with acute leukemias and multiple myeloma had a slower platelet recovery. Treatment related death was 4.5%. The status of the disease at diagnosis was the most significant prognostic factor. With a median follow-up of 23 months the probability of event-free survival at 60 months was 46% for low grade lymphoma, 44% for intermediate and high grade lymphoma, 58% for Hodgkin's disease, 45% for acute myeloblastic leukemia, 38% for solid tumors and 15% for multiple myeloma. The probability of survival at 60 months was 67% for low grade lymphoma, 47% for intermediate and high grade lymphoma, 75% for Hodgkin's disease, 52% for acute myeloblastic leukemia, 54% for solid tumors and 25% for multiple myeloma. It can be concluded that autologous progenitor cell transplantation induces a complete and faster hematological recovery in all groups of patients without any late graft failure. Results are similar to those published in the literature. The treatment-related death was low and acceptable. PMID- 10835709 TI - [Application of a preoperative evaluation model in patients undergoing elective abdominal surgery]. AB - We used a preoperative evaluation model based on the clinical history in order to determine the clinical characteristics of the patients. We also estimated the prevalence of asymptomatic disease in ASA1 patients and described the frequency of diagnosis that motivated suspension of the surgery during the preoperative time. Another purpose was to establish the perioperative complications according to the risk index and to detect the medical reasons for suspension of surgery during hospitalization. A total of 777 patients having an indication of surgery were evaluated in this program. Only 507 completed the preoperative evaluation. Of these, 57.79%, had at least one known disease or were diagnosed by their clinical history. Preoperative tests were indicated according to age in ASA1 patients. Complementary studies were indicated in relationship to the history and physical examination in ASA2 and ASA3 patients. After the evaluation, 27 surgeries were suspended: 21 were considered high risk, 2 suffered unknown active infections, and 4 ASA1 patients had severe asymptomatic anemia. A total of 328 patients were admitted for surgery. In 5 of these patients the anesthesiologist stopped the procedure in the operating room. The causes were acute decompensations of known pathologies. There were no complications or deaths related to unknown diseases and no patients died from cardiopulmonary or metabolic complications. In ASA1 patients, there were no complications related to this evaluation. This study allowed us to determine the clinical status of the patients and in consequence high-risk surgeries were canceled. In admitted patients a few surgeries were canceled for clinical reasons. This program probably decreased patient morbimortality, unnecessary hospitalization and costs. PMID- 10835710 TI - [Fluid overload in critically ill patients. Hemodynamic effect vs hydrosaline balance]. AB - The administration of high volumes of fluids during resuscitation of critically ill patients follows the recommendations of experts' committees that postulate volume expansion to correct deficits that may be associated with certain conditions (sepsis, trauma, surgery, etc), not primarily characterized by hypovolemia. These recommendations suggest that optimizing cardiac output may avoid occult deficit in tissue oxygen delivery and consumption that could lead to multisystem organic failure and death. The lack of benefit in the morbidity and mortality rates of patients with treatments directed to reach supranormal values of oxygen delivery, attempts against this hypothesis, since it fails to correct the primary hemodynamic dysfunction (arterial vasodilatacion) and generates fluid overload. The probable mechanisms linked with the avidity to retain water and sodium by critically ill patients are reviewed and the use of restricted protocols that preserve fluid balance is encouraged to potentially improve outcome. PMID- 10835711 TI - [Present and future of oral anticoagulants]. AB - The number of patients under oral anticoagulant therapy has markedly increased lately, mainly due to those with chronic atrial fibrillation. Progress has been made in the control of oral anticoagulation because sensitive and calibrated commercial reagents for prothrombin time have become available. But bleeding is still a problem in these patients. In our experience, the intensity and the duration of the anticoagulant therapy are the most important risk factors for bleeding. The deviation of INR (International Normalized Ratio) can also be associated with higher risk for bleeding. The limitations of oral anticoagulant therapy include frequent laboratory controls for dose adjustment, drug interactions, narrow therapeutic range and the high variability in patient response. These limitations prompted the development of new antithrombotic agents. A number of low molecular weight active site inhibitors of thrombin are being developed and one of them is orally bioavailable, and could become an alternative to vitamin K antagonists. PMID- 10835712 TI - [Antisense oligonucleotides increase the apoptotic effect of idarubicin in K-562 cell line]. AB - The cell line K-562, which carries bcr/abl rearrangement of type b3a2 is resistant to apoptosis induced by topoisomerase II inhibitors. K-562 cells were treated with complexes of cationic liposomes (DMRIE-DOPE and Dcchol-DOPE) and antisense oligonucleotides (AS-ODNs) directed against the b3a2 type of bcr/abl mRNA and non sense oligonucleotides (NS-ODNs), in a 3:1 lipid/DNA ratio during 72 hours, then they were incubated for a further 24 hours with idarubicin (IDA), 0.5 microgram/ml, to induce apoptosis. It was evaluated by morphology to the microscope of fluorescence. Cells treated with the complexes DMRIE-DOPE and Dcchol/DOPE with the specific AS-ODN showed a higher apoptosis percentage induced by IDA (mean +/- SD: 14.74 +/- 2.07 and 20.43 +/- 4.58, respectively) compared with controls not treated with ODNs (mean +/- SD: 8.08 +/- 0.82); (p < 0.05). These data indicate that the AS-ODNs directed against the b3a2 type of bcr-abl mRNA renders the cell line K-562 sensitive to IDA at the mentioned concentration. PMID- 10835713 TI - [Anatomy lessons]. PMID- 10835714 TI - [The concept of University according to Eduardo Braun Menendez]. PMID- 10835715 TI - [Molecular epidemiology of the human immunodeficiency virus type 1 in Santiago, Chile]. AB - BACKGROUND: Most of the studies of HIV-1 infection in South America have been limited to Brazil and little is known about the viral variants that are causing disease elsewhere in the continent. AIM: To determine the characteristics of the viral variants present in Chile as well as patterns of viral transmission. MATERIAL AND METHODS: Viral sequences were obtained from 21 HIV-1 infected people from Santiago, Chile who were infected either via sexual contact or intravenous drug use. Cloned sequences obtained from both the third variable and conserved regions of the envelope as well as the viral protease were evaluated. RESULTS: We found only clade B subtype viruses in Santiago. An evaluation of the envelope gene revealed no evidence that the sequences were monophyletic by risk group. A number of the protease sequences were predicted to encode amino acid substitutions commonly found during selection for protease inhibitor resistance. CONCLUSIONS: The HIV-1 strains studied in Chile, belong to the subtype B. There is no molecular evidence of separate introductions of the virus into the different risk groups. A number of substitutions in the protease gene that may confer resistance to protease inhibitors were found in patients with no previous exposure to this class of drugs. PMID- 10835716 TI - [In vitro immunosuppressive effect of low density lipoproteins]. AB - BACKGROUND: Immune cells participate in the formation of atheromatous plate, however little is known about the effects of native or oxidatively modified lipoproteins on these cells. AIM: To study the effects of lipoproteins on in vitro mononuclear cell proliferation. MATERIAL AND METHODS: Peripheral blood mononuclear cells were obtained from 10 patients with type 2 diabetes mellitus (aged 52 +/- 9 years old with a disease duration of 8.2 +/- 5.7 years and a mean glycosilated hemoglobin of 9.3 +/- 2.2%) and 10 non diabetic healthy controls (aged 50.3 +/- 7.1 years old). These were stimulated with phytohemagglutinin (PHA) alone or in the presence of native LDLS, malondialdehyde modified LDLs or glycated LDLs. Proliferation was measured as 3H-thymidine incorporation and expressed as Stimulation Index (SI). RESULTS: SI of patients and healthy subjects, after PHA stimulation were similar: (57.5 +/- 29.8 and 61.1 +/- 23.5) respectively LDLs did not induce proliferation in neither group. Native LDLs produced a 98% inhibition of PHA induced proliferation. Malondialdehyde modified and glycated LDLs caused a 50% inhibition. The suppressive effect was maintained when lipoproteins were incorporated to culture media 60 min prior or after PHA stimulation. CONCLUSIONS: Lipoproteins inhibit in vitro PHA induced peripheral blood mononuclear cell proliferation both in diabetic and in non diabetic subjects. PMID- 10835717 TI - [Nicotine dependence: 1-year follow-up patients treated with nicotine replacement and group therapy]. AB - BACKGROUND: Smoking is the main single avoidable cause of death in our country. Little research in the treatment of such disorder has been made. AIM: To report the results of a prospective follow up for one year of outpatients from our "Smoker's Clinic" at the Department of Psychiatry of the Catholic University. PATIENTS AND METHODS: One hundred twenty-seven patients (84 male, aged 21 to 70 years old), with DSM-IV criteria for nicotine dependence, were included in a total of 18 groups. Each group received an intensive treatment program of 10 sessions with cognitive-behavioral relapse prevention techniques and nicotine replacement. Patients with active psychiatric diseases were not included in the program. RESULTS: Eighty-seven percent of subjects were abstinent at the first month and 50% were still abstinent after twelve months of follow up. We did not find differences in gender, age, previous psychiatric disease, number of cigarettes and breath carbon monoxide level between abstinent and non abstinent patients after 12 months of follow up. CONCLUSION: This intensive nicotine dependence treatment in seriously dependent patients, proved to be successful, regardless of the previous psychiatric history. PMID- 10835718 TI - [Clinical and endoscopic findings and magnitude of gastric and duodenal reflux in patients with cardial intestinal metaplasia, short Barrett esophagus, compared with controls]. AB - BACKGROUND: The diagnosis of patients with short segments of intestinal metaplasia in the distal esophagus, has increased in recent years. AIM: To assess the clinical, pathological and functional features of patients with esophageal intestinal metaplasia. PATIENTS AND METHODS: A prospective study was performed in 95 control subjects, 115 patients with cardial intestinal metaplasia and 89 patients with short Barret esophagus with intestinal metaplasia. All had clinical and endoscopic assessments, esophageal manometry and determination of 24 h esophageal exposure to acid and duodenal content. RESULTS: Control patients were younger and, in this group, the pathological findings in the mucosa distal to the squamous-columnar change, showed a preponderance of fundic over cardial mucosa. In patients with intestinal metaplasia and short Barret esophagus, there was only cardial mucosa, that is the place where intestinal metaplasia implants. Low grade dysplasia was only seen in the presence of intestinal metaplasia. Gastroesophageal sphincter pressure decreased and gastric and duodenal reflux increased along with increases in the extension of intestinal metaplasia. CONCLUSIONS: These findings confirm the need to obtain multiple biopsies from the squamous-columnar mucosal junction in all patients with gastroesophageal reflux symptoms, for the detection of early pathological changes of Barret esophagus and eventual dysplasia. PMID- 10835719 TI - [Domestic violence and sexuality]. AB - BACKGROUND: Women's reproductive health is closely related to the conditions in which intercourse is carried out with the sexual partner. Physical and emotional abuse is expressed as loss of bonds, reduction in self esteem and deterioration in interpersonal relationships. AIM: To assess the effects of domestic violence on pregnant women attitudes towards sexual relations with their partners, and to critically analyze the dependence of these variables on biosociodemographic factors, socioeconomic status and family functioning. MATERIAL AND METHODS: Sixty three pregnant women with a domestic violence background (index group) and 43 women without such background (control group) were subjected to a structured survey including questions about biosociodemographic variables, socioeconomic status, family relations, couple interactions and questions about attitudes towards sexual relations. RESULTS: Domestic violence was caused by the woman's partner or ex partner in 73% of cases. A greater family dysfunction and disturbance in the couple's relation was observed in the index group (p < 0.05). Association tests and multiple correspondence analysis did not show a clear correspondence between variables related to a woman's attitude towards sex relations and the risk of domestic violence. CONCLUSIONS: This quantitative methodology was unable to identify the reasons, interpretations and meanings that women with a background of violence, give to features associated with their sexuality. PMID- 10835720 TI - [Systemic inflammatory response syndrome: is it comparable with severe sepsis?]. AB - BACKGROUND: In 1992, a consensus conference defined the terms systemic inflammatory response syndrome (SIRS), sepsis, severe sepsis and septic shock. Since then, numerous reports have validated the prognostic usefulness of these operative definitions. AIM: To evaluate if sepsis severity criteria, as defined by the Consensus Conference, can be applied to noninfectious SIRS. PATIENTS AND METHODS: Five hundred eighteen patients admitted to 5 intensive care units (ICU) from 4 hospitals were prospectively evaluated during a 3 months period. Patients that met at least one severity criteria were included. SIRS etiology, organ dysfunction and evolution were recorded in each patient. RESULTS: One hundred two patients were included: 79 with sepsis (group I) and 23 with noninfectious SIRS (group II). ICU and hospital mortality were comparable (43 and 48% in sepsis compared to 43 and 51% in non infectious SIRS). The most common sources of sepsis were pneumonia and peritonitis. Group II patients had a wide variety of diseases. ICU stay, APACHE score and number of organs with dysfunction were not different among groups. Only the incidence of renal dysfunction was higher in the septic group. CONCLUSIONS: The Consensus sepsis severity criteria can be applied to noninfectious SIRS, defining a population subset with similar high mortality and organ dysfunction incidence, although with greatly heterogeneous etiologies. PMID- 10835721 TI - [Conservative treatment of ductal carcinoma in situ of the breast]. AB - BACKGROUND: Since the widespread use of mammography, the incidence of ductal carcinoma in situ of the breast has increased. Until few years ago the standard treatment was mastectomy, however, from the analysis of conservative treatment trials for invasive carcinoma, it was evident that ductal carcinoma in situ could also be treated conservatively. This was confirmed later by randomized trials. AIM: To analyze the experience of our Institution with conservative treatment of ductal carcinoma in situ of the breast. PATIENTS AND METHODS: A search through the data base of our Institution found 69 patients treated with lumpectomy and radiotherapy between the years 1976 and 1997. RESULTS: Twenty three of 69 patients (33%) were diagnosed because of a palpable mass. Eleven of twelve were diagnosed prior to 1990 and 12 of 57 after 1990. With a median follow-up of 48 months local control and overall survival is 97%. None of the patients underwent mastectomy. CONCLUSIONS: Conservative treatment of ductal carcinoma in situ of the breast is a reasonable alternative, mainly if we realize that with increasing frequency--the diagnosis is made through mammography and with non-palpable lesions. The results reported in this study are similar to those reported by other centers. PMID- 10835722 TI - [Multidisciplinary management of malignant hepatic tumors in children: a recent national experience]. AB - BACKGROUND: Malignant hepatic tumors (Mht) are rare in children. Among them hepatoblastoma (HB) is the most common. AIM: To report the results of the multidisciplinary management in 6 consecutive children: five HB and one metastatic Wilms tumor (MWT). PATIENTS AND METHODS: The mean age of patients was 42 months. All HB patients had elevated serum alfafetoprotein (median 150,000 ng/ml). All patients received preoperative chemotherapy: HB patients received carboplatin/doxorubicin alternating with cisplatin, and the MWT patient, vincristine alone. Surgery included two formal right and two formal left hepatectomies, one extensive central resection with partial left segmentectomy, and one lateral segmentectomy. Extracorporeal circulation was used in the child with atrial involvement. All patients received postoperative chemotherapy. RESULTS: All tumors had variable regresion on preoperative chemotherapy. Complete resection with negative margins was achieved in all patients. The degree of tumor necrosis on histology ranged from 60% to 90%. Alfafetoprotein levels fell to under 10 ng/ml in all HB cases, one to three months after surgery. All patients survive free of disease at a median follow up of 19 months. CONCLUSION: A multidisciplinary approach including the well timed used of chemotherapy and surgery is highly effective in the management of pediatric malignant tumors. PMID- 10835723 TI - [Outbreak of acute histoplasmosis in Chilean travelers to the ecuadorian jungle: an example of geographic medicine]. AB - Eight Chilean teenagers traveled to Ecuador in January 1999, where they were bitten by mosquitoes, had contact with parakeets and lodged in poorly hygienic places; 6/8 visited for 5-10 minutes the interior of a bat cave. About a week later these 6 began with headache, myalgia and fever that lasted 2-3 weeks. 5/6 had dry cough with no respiratory distress. The index case was seen in the 2nd week of symptoms. A chest x-ray showed multiple nodular infiltrates as in the other five. Two had histoplasma serology, one was negative and the other positive at a low titer; histoplasmin skin test showed induration of 17-27 mm in all six. An acute histoplasmosis with massive exposure was diagnosed and treated with itraconazole for 3 weeks. All became asymptomatic and chest x-rays returned to normal. Histoplasmosis (non existent endogenously in Chile) is, among other geographic and tropical diseases, a risk for Chilean travelers. Awareness of this in the general population and development of expertise in these diseases by local health care providers is required. PMID- 10835724 TI - [Hemorrhagic tuberous sclerosis. Report of a Jehovah Witness patient]. AB - We report a 26 years old male with a tuberous sclerosis with multiple and bilateral kidney cysts and angiomyolipomas. The patient presented to the emergency room with a severe abdominal pain and anemia, secondary to a bleeding angiomyolipoma. The patient rejected blood transfusions due to his religious beliefs. A selective angiography was performed confirming diagnosis and the lesion artery was selectively embolized, stopping the bleeding immediately. The patient had a satisfactory evolution thereafter. This is a rare lesion and the fact that the patient was a Jehovah witness that rejected blood transfusions, required an innovative medical approach. PMID- 10835725 TI - [Segmental short bowel stenosis after a superior mesenteric vein thrombosis. Report of a case]. AB - We report a 24 years old female with a Superior Mesenteric and Portal Vein Thrombosis due to an Antithrombin III factor deficiency, associated to oral contraceptive use and smoking. She presented with severe abdominal pain and the diagnosis was reached after surgery with a CT scan. The patient was treated with intravenous heparin and oral anticoagulation, with a good clinical and Doppler endo-sonographic response. One month after the onset, she developed an intestinal occlusion caused by two concentric jejunal stenoses, measuring 2 and 0.7 cm in length and demonstrated with a barium jejunogram. A 35 cm intestinal resection was done and the patient recovered uneventfully. The pathological study showed granulation tissue on both stenotic zones with an ulcer near to the distal stricture, that reached the internal muscularis propria, with subserosal fibrosis. The development of segmental stenosis is a rare complication superior mesenteric vein thrombosis, that must be bore in mind. PMID- 10835726 TI - [Professional quality assurance. A new ethic frame for the practice of medicine]. AB - Accreditation of physicians and medical education programs are progressively standing as well established procedures in most related institutions and countries, worldwide. Yet, an ample and consistent review of this issue is still required for a more complete knowledge and universal acceptance. Quality health care is a goal of the profession and a demanding requirement of society. Reaching high standards in medicine is a major responsibility of the medical schools and of the pertinence and appropriateness of the programs. This is a useful and sound frame for the model that is being developed in Chile, to better regulate and improve both medical education and practice. The bioethics implications that merge from this perspective are underlined to give the accreditation system a deep moral relevance as are expression of people's wills and expectations on medical professional services. PMID- 10835727 TI - [Factors that affect DNA content analysis by flow cytometry]. AB - DNA ploidy and cell cycle analysis by flow cytometry is used to obtain additional information about the diagnosis and prognosis of different types of cancer. However, there are several disagreements among authors about the tissue source (fresh-frozen or paraffin embedded), cellular dissociation methods (mechanical, enzymatic or other), use of different dyes, lasers, analysis software with different mathematical models and interpretation of results. A discussion about the different aspects that affect the study of DNA ploidy and cell cycle and a consensus in publications is mandatory. A strict control of analysis processes and data interpretation is also necessary. PMID- 10835728 TI - [Evidence based medicine: a strategy that moves closer to or away from internal medicine?]. AB - In the last years, evidence based medicine has been considered a main factor for the diagnostic and therapeutic decisions in medical practice. However, one must consider the complexity of human beings and the difficulties in the interpretation of some results obtained in randomized prospective trials that support this theory. Therefore the importance of clinical experience, rational and logical medical thinking and facing patients as individuals must prevail at the moment of complex medical decisions. PMID- 10835729 TI - [Recommendations for the use of Sildenafil in patients with cardiac diseases]. PMID- 10835730 TI - [The hepatitis viruses: from A to TT]. AB - A great progress in the knowledge about hepatitis virus has occurred in the last decades and several virus have been identified. Virus B, C and D are transmitted parenterally, while virus A and E use the enteral route. Highly effective vaccines for A and B hepatitis are presently available. Virus C is an important cause of chronic liver disease at the present moment. Since the identification of virus C and E, the number of acute hepatitis denominated as non A non B has decreased considerable. Although there are still patients with viral hepatitis of unknown origin and there is considerable effort to identify the agents causing them. Virus G and TT are frequently present in the sera of patients with chronic liver diseases but their real pathogenic role is not completely elucidated. PMID- 10835731 TI - [Current status of immunity to hepatitis A virus in various adult groups]. AB - BACKGROUND: As sanitary and economic conditions improve, the prevalence of antibodies to hepatitis A is now significantly lower. AIM: To evaluate the prevalence of hepatitis A virus antibodies in healthy Chilean adults. MATERIAL AND METHODS: Antibodies to hepatitis A virus were measured, using a commercial ELISA assay, in 215 voluntary blood donors (163 male, aged 19 to 30 years old) and 295 medical students and health personnel (156 male, aged 19 to 39 years old), residing in Valdivia, Chile. RESULTS: Antibodies against hepatitis A virus were found in 68.2% of the total sample (351/510). Ninety percent of flood donors and 54% of health personnel and students were positive (p < 0.01). Age specific prevalence in blood donors 19 to 22, 23 to 29 and 27 to 30 years old was 81.0%, 95.2% and 95.6% respectively. Among the same age groups in medical students, the prevalence was 47.9%, 53.2% and 61.9% respectively (p < 0.01). CONCLUSIONS: This study indicates a reduction in the prevalence of hepatitis A virus antibodies among adults in Valdivia (Chile). Differences detected between individuals are probably related to different socioeconomic levels. Medical students have an increased risk for hepatitis A infections than the general population. PMID- 10835732 TI - [Prevalence of obesity, hypertension and dyslipidemia in rural aboriginal groups in Chile]. AB - BACKGROUND: Chilean aboriginal ethnic groups (mapuche and aymaras) have a very low prevalence rate of type 2 diabetes. The investigation of a possible relationship between this low prevalence of diabetes and obesity, hypertension and serum lipid profiles in both groups is worthwhile. AIM: To study the prevalence of obesity, hypertension and lipid profile in two Chilean aboriginal communities. SUBJECTS AND METHODS: The prevalence of obesity, hypertension, fasting serum total cholesterol, HDL cholesterol, triglycerides, glucose, insulin, leptin and oral glucose tolerance test were measured in 345 mapuche (106 male) and 247 aymara (100 male) individuals. RESULTS: Sixty three percent of mapuche women, 37.9% of mapuche men, 39.7% of the aymara women and 27.0% of aymara men had a body mass index over 27 kg/m2. Twenty percent of mapuche men, 18.0% of mapuche women, 9.0% of aymara men and 4.8% of the aymara women had high blood pressure values. Serum HDL cholesterol was below 35 mg/dl in 16% of mapuche women, 14% of mapuche men, 25% of the aymara women and 27% of aymara men. No differences in total cholesterol levels were observed between mapuches and aymaras. CONCLUSION: Mapuche women have higher prevalence of obesity and high blood pressure than aymara women. Low serum HDL cholesterol has a higher prevalence among aymara individuals. PMID- 10835733 TI - [Value of intestinal manometry in gastrointestinal pathology]. AB - BACKGROUND: Small intestinal manometry is a relatively simple technique. However, its use is usually limited to very few centers and mainly related to research studies. AIM: To report our experience with small intestinal motility studies in a group of normal controls and patients with symptoms suggesting a gastrointestinal motor disorder. PATIENTS AND METHODS: Seventy three studies were performed in 71 subjects: 18 asymptomatic controls and 55 patients presenting with symptoms characterized by abdominal pain, vomiting, bloating, constipation and diarrhea. In 33 patients the same symptoms remained without diagnosis, in spite of extensive laboratory studies. In 10 of these, dilated small intestinal loops were observed and intestinal pseudoobstruction was suspected. Twenty two additional patients with systemic disorders such as scleroderma, diabetes and previous vagotomies, were studied. Motility was assessed by means of perfused catheters connected to external transducers for a mean lapse of 280 min. RESULTS: An abnormal pattern of small intestinal motility suggesting neuropathic, myopathic or a mixed disorder was observed in 76% of all studied patients, with the exception of patients with scleroderma, in whom only myopathic and mixed alterations were observed. In 82% of patients, the results of manometry were useful for the management of the clinical condition. CONCLUSIONS: Small intestinal manometry is a relatively simple technique that, when used in selected groups of patients, provides useful information for clinical management. PMID- 10835734 TI - [Availability of physicians in Chile and its medium-term projection. 5 years later]. AB - According to a prospective study performed in 1993, Chile had 14,400 active physicians (excluding those over 70 years of age) and a physician/inhabitant ratio of 1:960. The estimated projection for 1998 was of 16,244 physicians and a ratio of 1:921. However, during the present decade, the number of medical schools in Chile has duplicated, increasing by 38.4% the number of available posts in Medical schools. Also, 1,297 physicians graduated abroad, have been incorporated. Therefore, previous projections must be revisited. The present study shows that in 1998, Chile has 17,441 physicians (1:850), will have 20,610 (1:765) in 2003 and 24,449 in 2008. This last figure means a physician/inhabitant ratio of 1:679, similar to that of countries with a consolidated market economy. The foreseen medical population increase rate of 3.5% doubles the general population increase rate. To assure the quality of medical training, a Medical School accreditation process is being held. This study highlights the need to review bilateral agreements subscribed with five countries, that recognize the physician degree without assessing the medical skills of foreign graduates. PMID- 10835735 TI - [Association of non-syndromic cleft lip and cleft palate with microsatellite markers located in 6p]. AB - BACKGROUND: Nonsyndromic cleft lip with or without cleft palate (NSCLP) is a common craniofacial developmental defect. Association studies have suggested that a clefting locus is located on chromosome 6p at or near two possible loci, Factor 13A (FI3A) in the region 6p 25-24 and HLA at 6p 21.3. AIM: To test the hypothesis on the possible presence of a major gene on chromosome 6p associated with NSCLP. PATIENTS AND METHODS: We carried out an association study on a sample of unrelated NSCLP patients from multiplex (Mx) and simplex (Sx) families, of their unaffected relatives and in control individuals. DNA was analyzed with three PCR markers close to the putative NSCLP locus, dinucleotide repeats at loci D6S89, D6S109 and D6S105. PCR products were resolved by PAGE and visualized by silver staining. Statistical analysis was performed by means of chi 2 log ratio. RESULTS: Significant differences were observed when comparing the allele frequency distribution of D6S89 in patients with NSCLP and controls and in patients with NSCLP-Mx and controls. No significant differences were observed for patients with NSCLP-Sx. D6S109 and D6S105 showed no significant differences in any of the comparisons. CONCLUSIONS: Our results support the hypothesis that a NSCLP locus maps on 6p23 very close to D6S89. Results for D6S109 and D6S105 do not show a clear association. Differences observed between NSCLP-MX and Sx families seem to represent different etiologic entities. The results of the present study, plus those already published for candidate loci, TGFA and MSX1, support the hypothesis that several interacting major genes participate in the etiology of NSCLP. PMID- 10835736 TI - [Frozen-section biopsy in ovarian neoplasm diagnosis: diagnostic correlation according to diameter and weight in tumors of epithelial origin]. AB - BACKGROUND: Adequate management and treatment of ovarian carcinoma requires a complete surgical staging supported by frozen-section examination. To achieve this goal it is necessary a high level of accuracy. AIM: To evaluate the accuracy of frozen-sections in ovarian carcinoma considering the influence of tumor diameter and weight. PATIENTS AND METHODS: Retrospective study of frozen-sections performed in patients with ovarian tumors who underwent surgery. Frozen- and permanent-sections were divided into three categories (benign, borderline and malignant) and stratified by diameter (< 10 cm, 10 to 20 cm, > 20 cm) and weight (< 700 g, 700 a 1400 g, > 1400 g). The diagnostic correlation, sensitivity, specificity, predictive values and accuracy of each frozen-section diagnosis were determined. RESULTS: Eight hundred forty two ovarian tumors that underwent frozen sections between January 1988 and October 1998 were studied. Final diagnosis was 86.7% benign, 2.7% low malignant potential (LMP) and 10.6% malignant. The diagnosis correlation between frozen- and permanent-sections was 98.2%. Misdiagnosis was in epithelial ovarian tumors, particularly in LMP tumors. Sensitivity, specificity, positive- and negative-predictive values and accuracy of the four hundred eighty nine epithelial tumor were 92.6%, 99.2%, 96.7%, 98.2% and 97.9%, respectively. Diagnostic correlation was higher in epithelial ovarian tumors with diameter < 10 cm (98.2% v/s 93.8%) and weight < 700 g (96.9% v/s 88.9%). CONCLUSIONS: Diagnostic correlation with permanent-section examination, sensitivity, specificity and predictive values of frozen-sections are high in ovarian tumors. Accurate diagnosis at frozen sections of epithelial ovarian tumors with diameter > 10 cm or weight > 700 g (particularly in LMP tumors) is difficult because of the extensive sampling required. Frozen-sections diagnoses are important to determine the type and extent of surgery performed at the initial operation. PMID- 10835737 TI - [Colorectal neoplasms. Treatment of obstruction with autoexpanding metal stents]. AB - BACKGROUND: Expandable metal stents can be used as a palliative or pre surgical method to decompress obstructing colonic carcinomas. AIM: To assess the effectiveness of these stents in the treatment of obstructive colonic carcinoma. PATIENTS AND METHODS: Expandable metal stents were placed in nine patients with the diagnosis of colorectal carcinoma and with clinical and radiographic signs of intestinal obstruction. Stents were placed under fluoroscopic guidance in nine patients and with endoscopic help in 2. The indications were palliative treatment in 7 and pre surgical decompression in 2 patients. RESULTS: Stent placement was successful in all patients. One patient presented a self limited rectal bleeding after the procedure. Obstruction was relieved in less than 24 hours after the procedure. CONCLUSION: Expandable metal stent placement is an effective means of relieving intestinal obstruction caused by colorectal carcinoma. PMID- 10835738 TI - [Construction and validation of a gastroesophageal reflux symptom scale. Preliminary report]. AB - BACKGROUND: Gastroesophageal reflux symptoms are frequent in the general population, but there is a lack of information about its prevalence and valid and reliable scales to determine it. AIM: To design a valid and reliable scale to determine gastroesophageal reflux disease. SUBJECTS AND METHODS: A Cross sectional study with patients and controls was conducted. The diagnosis of gastroesophageal reflux was based of clinical plus radiological, endoscopic or pathologic criteria. A structured questionnaire was built, containing items covering different clinical manifestations of gastroesophageal reflux disease. All subjects were subjected to 24 h esophageal pH monitoring. Internal consistency of items, interobserver reliability, criterion and construct validity, sensitivity, specificity and predicted values were obtained. RESULTS: One hundred thirteen subjects (74 female and 39 male with a mean age of 46 years old) were studied. Seventy three had gastroesophageal reflux and 40 were controls. Internal consistency of the score was 0.82. Inter observer reliability was greater than 0.80 and a significant association was observed between the score and 24 h esophageal pH monitoring (p < 0.001). Instrument sensitivity was 97% and specificity 83%. CONCLUSIONS: The designed scale proved to be a valid and reliable instrument to measure gastroesophageal reflux symptoms, that can be used for future prevalence studies. PMID- 10835739 TI - [Severe and recent hyponatremia and hypokalemia associated to the use of hydrochlorothiazide, enalapril and citalopram. Clinical case]. AB - We report a 72 years old hypertensive female, treated with enalapril 10 mg/day and hydrochlorothiazide 25 mg/day during three years. She presented a depressive disorder and cytalopram was prescribed in a dose of 10 mg/day. Two weeks before admission, a serum electrolyte analysis disclosed normal results and the cytalopram dose was increased to 20 mg/day. The patient was admitted with a hyponatremic encephalopathy with a plasma sodium of 100 mEq/L and a plasma potassium of 2.0 mEq/L. Cytalopram, enalapril and hydrochlorothiazide were discontinued, hypertonic NaCl and KCl were administered. The patient had a favorable evolution with a remarkable improvement of her symptoms. PMID- 10835740 TI - [Treatment of refractory ascites using a transjugular intrahepatic portosystemic shunt (TIPS). Clinical case]. AB - Approximately, 10% of cirrhotic patients with ascites become refractory to medical treatment, a condition that reduces overall survival. TIPS is a therapeutic alternative for the management of refractory ascites. It corrects sinusoidal portal hypertension, becoming a rational and relatively safe therapeutic approach. The most frequent complications of TIPS are the development of hepatic encephalopathy and a high incidence of shunt stenosis or occlusion after one year of follow-up. We report a 43 years old alcoholic cirrhotic male with ascites refractory to medical treatment, that required multiple large volume paracentesis, whose clinical condition deteriorated progressively. A TIPS was successfully placed with a rapid and complete resolution of ascites and marked improvement of his clinical condition. During the ensuing six months the patient remained in excellent conditions, but was lost from follow-up and resumed excessive alcohol intake. Eight months after TIPS placement he had an upper gastrointestinal bleeding and died. PMID- 10835741 TI - [Massive epistaxis associated with trauma. An indication for superselective angiography and embolization. Clinical case]. AB - We report a 23 years old male admitted to the hospital after a fall from a great altitude with severe trauma. During the second week of hospitalization he presented a severe nosebleed that did not respond to conventional tamponade. A selective angiography was performed and the branches of the internal maxillary artery were embolized in two occasions, stopping the bleeding. The patient was discharged from the hospital after 71 days of hospitalization. PMID- 10835742 TI - [Hepatitis C virus and resulting diseases]. AB - In 1989, the main agent causing non A non B hepatitis was identified as a RNA virus of the flavivirus family, with several serotypes, and was denominated virus C. At the present moment, the knowledge about the infection features and diseases that it causes has expanded thanks to the availability of reliable laboratory techniques to detect the antibody and the virus. The prevalence of infection and the frequency of serotypes varies in different regions of the world. Chile is a country with a low prevalence. The detection of infected blood in blood banks has reduced the spreading of the disease. Other means of infection such as the use of intravenous drugs, hemodialysis and transplantation have acquired greater importance. Sexual, maternal and familial transmission is exceptional. Infected people develop an acute hepatitis, generally asymptomatic. Eighty percent remain with a chronic hepatic disease, that can be mild or progressive, evolving to cirrhosis or hepatic carcinoma. Chronic hepatitis, closely resembling an autoimmune disease, can be caused by the virus. Alcohol intake increases viral activity causing severe hepatic diseases, refractory to treatments. Several non hepatic diseases are associated to hepatitis C virus infection such as essential mixed cryoglobulinemia, mesangiocapillary glomerulonephritis, porphyria cutanea tarda, dysglobulinemias and probably type 2 diabetes mellitus. The only available treatment is interferon, that is successful in a minority of patients, frequently causing a transient improvement. The use of Ribaravine associated to interferon improve the effectiveness of therapy. Liver transplantation is the only therapy for severe hepatic disease. The use of new antiviral drugs should improve the prognosis of the disease. PMID- 10835743 TI - [Recent progress in the diagnosis of systemic vasculitis]. AB - The clinical picture of primary systemic vasculitis, that cause inflammation and necrosis of vessel walls, depend on the type, size and location of involved vessels. Frequently, their clinical presentation does not reflect the specific type of vasculitis and its prognosis. Moreover, the correct diagnosis, early and adequate treatment have an important prognostic value. Recently, new diagnostic methods for systemic vasculitis have been devised. This article critically reviews and provides information for the rational use of these new imaging techniques and laboratory procedures for the diagnosis and follow up of systemic vasculitis. PMID- 10835744 TI - [Claudio Costa Casaretto M.D. (1914-1999) in Revista Medica de Chile]. AB - The work of Dr. Claudio Costa Casaretto covers a broad field in the history of Medicine in Chile. He contributed with the historical aspects in the Centennial issue of Revista Medica de Chile in July, 1972. He published 85 papers in this journal in a lapse of 20 years. In his works, he investigated about medical personalities with the highest relevance for Chilean medicine such as Dr. William Blest, graduated in Edinburgh and director of the first Medicine Course in Chile in 1833, the French obstetrician Dr. Lorenzo Sazie, first Dean of the Faculty of Medicine of the University of Chile in 1843 and Dr. Eloisa Diaz, first physician graduated in Chile and South America in 1887. He published the translation from Latin of Juan Ignacio Molina's verses "Elegies to smallpox", Chilean writer and erudite of the XVIII century. He also undertook the origins of Universidad de San Felipe (1737), Universidad de Chile (1842) and Pontificia Universidad Catolica de Chile (1889) and the main educational events occurred in the Faculty of Medicine of the University of Chile, during the past century. He also published about the public health situation and sanitary care during the XIX century, about the conflict between private and public teaching and other political events of the past century. The work of Dr. Costa as a whole, is a real history textbook of Chilean Medicine. Dr. Costa and Dr. Enrique Laval are the most important Chilean medical historians of the XX century. PMID- 10835745 TI - [Recommendations for the use of ambulatory blood pressure monitoring. Consensus document of the Chilean Hypertension Society]. AB - Continuous ambulatory blood pressure monitoring is a diagnostic technique devised as a consequence of the great variations in blood pressure measurements. It allows multiple daily measurements, nocturnal monitoring, avoids the stress of blood pressure measurements, gives a picture of pressure behavior during 24 hours and reduces observer related errors. The equipment used must be accurate and validated using international protocols. Accepted indications for continuous ambulatory blood pressure monitoring are white coat hypertension, episodic hypertension, resistance to medications and assessment of symptoms or autonomic dysfunction. Other indications with less clear cut usefulness, are high risk cardiac, renal or pregnant patients and an accurate blood pressure control. We describe equipment calibration, elements that must be considered in the reports, result interpretation and conclusions. Normal blood pressure ranges for children and pregnant women are also reported. PMID- 10835746 TI - [Lithium and bipolar disorders]. PMID- 10835747 TI - [Heart disease, pregnancy and anticoagulant therapy]. AB - Pregnancy is a hypercoagulable state. Some women with cardiac disease and mechanical valve prosthesis are at increased risk of arterial thromboembolic phenomena. These women are maintained on oral anticoagulants and require effective permanent prophylaxis during pregnancy. The use of oral anticoagulants during pregnancy is controversial because of the risks of embriopathy (Chondrodysplasia punctata) in fetuses who are exposed to coumarin between the 6th and 9th week of gestation, the risk of neurological disorders all through pregnancy, and a higher incidence of abortion and stillbirths. The exact incidence of these complications is unknown. Most of this information comes from North American reports, when much higher mean daily doses of coumarin were administered, and they were probably overemphasised. Reports from Europe, Asia and our own, show that both embriopathy and central nervous system malformations are probably dose-related and that the risks of abnormality to the fetus are small. The ACC Antithrombotic Consensus (1998) recommends the use of subcutaneous heparin all through pregnancy or until the 13th week of gestation. Heparin does not cross the placenta, however there is a higher risk of maternal bleeding, abortion and stillbirths. With the use of small dose of heparin and of therapeutic heparin doses there is a risk of prosthesis thrombosis. Anticoagulant treatment of patients with prosthetic heart valves during pregnancy remains controversial. Subcutaneous heparin prophylaxis is feasible but the use of well controlled oral anticoagulants appear to offer lower risks of maternal and fetal complications. PMID- 10835748 TI - [Birth weight difference in 3 biological variables in normal newborns]. AB - BACKGROUND: The most important determinant of birth weight is gestational age. However, other biological variables may influence this parameter. AIM: To study the influence of maternal parity, height and neonatal gender on the birth weight of normal newborns in a public Hospital of Southern Chile. MATERIAL AND METHODS: A computer registry of all deliveries attended in Temuco Hospital between 1993 and 1998 was analyzed. Normal deliveries giving birth to newborns of more than 2,500 g and from women who did not suffer from conditions that could influence birth weight, such as hypertension, undernutrition or smoking, were selected. RESULTS: From 27,736 deliveries, 12,580 were selected for the study. Male newborns with gestational ages from 37 to 42 weeks, had a higher weight than their female counterparts. Multiparous women and those with a height over 154 cm gave birth to newborns with a higher weight than nulliparous women or than those with a height of less than 154 cm. CONCLUSIONS: According to these results, birth weight must be corrected by gender and maternal features, to define those newborns that have a low birth weight for their gestational age. PMID- 10835749 TI - [Association between nonsyndromic cleft lip/palate and microsatellite markers located in 4q]. AB - BACKGROUND: Nonsyndromic cleft lip with or without cleft palate (NSCLP) is a common craniofacial defect. Association studies have suggested that a clefting locus is located on chromosome 4q at or near two microsatellite markers D4S175 and D4S192. AIM: To test the hypothesis on the possible presence of a clefting locus on chromosome 4q. MATERIAL AND METHODS: We carried out an association study on a sample of unrelated NSCLP patients, of their unaffected relatives and in controls. Both probands and relatives were further analyzed depending if they originated from simplex or multiplex families. DNA was analyzed with two PCR markers close to the putative NSCLP locus, dinucleotide repeats D4S175 and D4S192. PCR products were resolved by PAGE and visualized by silver staining. Statistical analysis was performed by means of chi 2 log ratio. RESULTS: Significant differences between NSCLP and controls were observed when comparing the allele frequency distribution of D4S192 both in the total sample as well as in NSCLP-multiplex and simplex cases. No significant differences for D4S175 were observed in any of the comparisons. Unaffected relatives showed significant differences with controls both for D4S175 and D4S192. CONCLUSIONS: Our results support the hypothesis that a NSCLP locus maps on chromosome 4q close to the microsatellite marker D4S192. No differences were observed between NSCLP multiplex and simplex cases versus controls, implying that they do not represent different etiologic entities. The results of the present and previous studies in the same group of patients support the hypothesis that several major interacting genes participate in the etiology of NSCLP. PMID- 10835750 TI - [Prevalence of cardial or fundic mucosa and Helicobacter pylori in the squamous columnar mucosa in patients with chronic patological gastroesophageal reflux without intestinal metaplasia comparated with controls]. AB - BACKGROUND: The mucosa distal to the endoscopic mucosal change zone can have easily diagnosed early alterations, in patients with chronic gastroesophageal reflux. AIM: To determine the type of mucosa existent in the zone distal to the squamous-columnar junction in patients with chronic gastroesophageal reflux without intestinal metaplasia. PATIENTS AND METHODS: One hundred thirty four controls and 208 patients with chronic gastroesophageal reflux lasting two years were studied. Forty three of these patients had a normal endoscopy, 54 had an erosive esophagitis and 111 had a short columnar epithelium covering the distal esophagus, without intestinal metaplasia. In all subjects, four biopsies were obtained from a zone distal to the squamous-columnar junction and two from the distal gastric antrum. RESULTS: In 59% of control subjects, fundic mucosa was present in the zone distal to the squamous-columnar junction. Cardial mucosa was present in the rest. In patient with chronic gastroesophageal reflux, cardial mucosa was predominant. Helicobacter pylorii infection decreased along with increasing extension of cardial mucosa covering the distal esophagus. CONCLUSIONS: In patients with chronic gastroesophageal reflux there is a metaplasia of fundic mucosa towards cardial mucosa. On the other hand, Helicobacter pylorii infection decreases gradually. PMID- 10835751 TI - [Emergence of resistance to macrolides in Streptococcus pyogenes]. AB - BACKGROUND: Diseases produced by Streptoccocus pyogenes are still a problem in Chile, as in the rest of the world. It exhibits in vitro susceptibility to different antimicrobials, but penicillin continues to be the treatment of choice. Alternative drugs have been developed for allergic patients, such as erythromycin, new macrolides and cephalosporins. Nevertheless, resistant strains are appearing due to the indiscriminate use of macrolides. AIM: To assess present antimicrobial susceptibility of S Pyogenes strains isolated from chilean patients. MATERIAL AND METHODS: The susceptibility to penicillin, macrolides, clindamycin, cephalotin and vancomycin of 153 S Pyogenes strains, obtained from different health centers of the Metropolitan Region and isolated between 1996 and 1998, was assessed using the Kirby-Bauer method. Agar dilution minimal inhibitory concentration was then determined to macrolide resistant strains. RESULTS: All strains were susceptible to penicillin. There was a 7.2% cross-resistance to macrolides. CONCLUSIONS: These results confirm that S Pyogenes resistance to macrolides has increased considerably in the Metropolitan Region of Chile during the last years. PMID- 10835752 TI - [Occupational stress in health care personnel]. AB - BACKGROUND: Occupational stress results from the interaction of multiple risk factors, such as the physical environment, biological function disturbances, work content and organization and diverse psychosocial components. AIM: To study the frequency of symptoms and the main sources of job stress, perceived by professional and non professional health care workers and to compare gender differences. SUBJECTS AND METHODS: A group of specially designed, self administered, questionnaires adapted by the main author and independently validated, were applied. The results of two symptoms, one work satisfaction, and one job stress scales are reported. RESULTS: One hundred sixteen women and 89 men were studied, 143 are professionals and 62 non professionals (clerical and nurse auxiliaries). Forty percent had symptoms of job stress and 82 subjects were defined as "probable cases", according to the ratings on the symptom scales. There were significant differences in the frequency of symptoms between professionals and non professionals (34.3 and 69.2% respectively). The main differences in symptoms, sources of job stress and dissatisfaction were more closely associated to the occupational level than to gender. CONCLUSIONS: A high frequency of job stress symptoms was observed in this sample of health care workers and the risk of occupational stress varies more with the occupational level than with gender. PMID- 10835753 TI - [Dental fluorosis: quantification of Streptococcus mutans in school children from Mamina, Chile. A longitudinal study]. AB - BACKGROUND: High fluorine concentrations in drinking water are associated with a decrease in the amount of salivary S mutants. Taking into account that clinical dental fluorosis can appear with 1.5 ppm of fluorine in the drinking water, fluorine concentrations in Mamina is 2.4 ppm. AIM: To quantify salivary S mutans in school age children from Mamina, a zone with a high fluorine content in the drinking water, during one year. MATERIAL AND METHODS: During 1997 and 1998, dental health was assessed and salivary samples were obtained from 51 children (27 male) aged 10 +/- 2 years to quantify S mutans. RESULTS: Most children studied had more than 10(5) salivary S mutans colony forming units. No changes in the rates of infection or dental health characteristics were observed during the observation year. CONCLUSIONS: High fluorine content in the drinking water did not have an effect on salivary S mutans infection in this population. PMID- 10835754 TI - [Anatomical and pathological study of the temporomandibular joint disk in Colombian individuals]. AB - BACKGROUND: The temporomandibular joint is an oval fibrous plate that completely divides the joint and accommodates irregularities existent in the bone articular surfaces. AIM: To study the frequency of temporomandibular joint disk abnormalities among Colombian subjects. MATERIAL AND METHODS: We studied 120 temporomandibular joint disks obtained from fresh cadavers arrived in the Instituto de Medicina Legal, in Cali Colombia. These were analyzed according to subject's dental status and age. RESULTS: Nineteen percent of disks had small erosions. In 7.5% of disks there were wider erosions or perforations. Twenty six percent of discs were very thin and this alteration was seen mostly in people aged 50 years or older. CONCLUSIONS: These results are intermediate between those who claim that disk perforations are rare and those who had shown tile opposite. The loss of dental pieces seems to be a risk factor leading to such disk perforation. PMID- 10835755 TI - [Subcutaneous heparin during the first trimesters of pregnancy in women with prosthetic heart valves]. AB - BACKGROUND: Oral anticoagulation therapy in pregnant women with prosthetic heart valves is associated with a greater risk of abortion, fetal malformations and thromboembolic complications. AIM: To assess the use of subcutaneous heparin in women with pregnancies of less than 12 weeks as an alternative to oral anticoagulation therapy. MATERIAL AND METHODS: Pregnant women were admitted to the hospital and coumarinics were replaced with not fractionated subcutaneous heparin in a dose of 10,000 IU every 12 hours, aiming at prolonging partial thromboplastin time, 2 or 2 1/2 times. During the hospitalization period, women were taught about the heparin injection technique. Coumarinics were restarted after week 13 of pregnancy until 10 days prior to the delivery date in which women were again admitted to the hospital and intravenous heparin was used until the delivery. RESULTS: Between 1991 and 1997, this protocol was used during ten pregnancies in seven women aged 19 to 36 years old. Five had a Starr-Edwards prosthesis, one had a mitral Bjork-Shilley prosthesis and one, a double prosthesis (mitral Starr-Machi and aortic St Jude). Subcutaneous heparin was started in the fifth week of pregnancy in 1 case, in the sixth week in seven and in the seventh week in two. There was no maternal mortality and one transient ischemic attack without sequelae. One non compliant patient had a Bjork Shilley prosthetic valve dysfunction that required a valve replacement at the twelfth week of pregnancy and she had a spontaneous abortion at week 15. Other patient had a pneumonia at week 37 and gave birth to a stillbirth. There were no fetal malformations. CONCLUSIONS: This therapeutic protocol can be used among Chilean patients, but must be restricted to compliant women and a strict surveillance must be maintained. PMID- 10835756 TI - [Tics disease (Gilles de la Tourette syndrome): clinical characteristics of 70 patients]. AB - BACKGROUND: Tourette's syndrome is a childhood-onset hereditary neurobehavioural disorder believed to occur without geographical restrictions. Although there have been reports of this disorder worldwide just a few are from Latin America. AIM: To report a preliminary experience with a series of 70 patients and to review recent advances in this disorder. PATIENTS AND METHOD: We reviewed patients seen in pediatric and adult neurological clinics in Santiago, Chile, all of whom fulfilled clinical diagnostic criteria for Tourette Syndrome. RESULTS: Seventy patients were studied, 54 males (77.1%) and 16 females (22.8%), their mean age at first evaluation was 13.6 years (range 2-46). The mean age of onset of symptoms was 6.4 (range 2-20), the mean time of follow-up was 3 years. Fifty-eight patients showed simple motor tics (blinking, facial grimacing, shoulder shrugging), whereas dystonic tics like head jerking were seen in 38 patients, torticollis in 6 and oculogyric movements in 2. Complex motor tics like jumping, antics, trunk bending and head shaking were present in 16 subjects. Vocal tics were predominantly of the simple type: sniffing, throat clearing, blowing, and whistling. Complex vocal tics were seen in 12 patients, five cases showed palilalia, 3 echolalia and only six displayed coprolalia (8.5%). Tics were of mild to moderate severity in most patients. Obsessive-compulsive disorder was observed in 22.8% and attention deficit and hyperactivity disorder were present in 35.7%. Forty-five patients (64.2%) had a first degree relative with tics, nine patients (12.8%) had a family history of obsessive-compulsive disorder. The current evidence involving desinhibition of cortico-striatum-thalamic-cortical neuronal circuits in the pathogenesis of this disorder is analyzed. CONCLUSION: Our report supports the recognized clinical homogeneity and genetical basis of Tourette's syndrome regardless of geographical region and ethnic origin. PMID- 10835757 TI - [Hepatotoxicity by amoxicillin/clavulanic acid: case report]. AB - We report a 72 years old diabetic male that, after the use of combined amoxicillin-clavulanic acid, developed pruritus and jaundice. Liver function tests showed serum total bilirubin of 4.3 mg/dL aspartate aminotransferase 140 U/l (normal < 35 U/L), alanine aminotransferase 470 U/L (normal < 40) and alkaline phosphatases of 400 U/L (normal < 100). Serology for hepatitis A, B and C viruses was negative, ERCP showed a normal biliary tree and liver biopsy disclosed a cholestatic hepatitis. Ursodeoxycholic was started to relieve pruritus. Liver function tests improved shortly thereafter, suggesting that this drug may be useful in the treatment of drug induced cholestasis. PMID- 10835758 TI - [Obstructive jaundice in prostate neoplasm. Case report]. AB - We report a 58 years old male, presenting with malaise, weight loss and jaundice. An abdominal ultrasound showed multiple lymphadenopathies in the hepatic bilus and around the pancreas. Fine needle aspiration of these nodes demonstrated an undifferentiated carcinoma. Prostate specific antigen was over 100 ng/ml and a prostate biopsy demonstrated a high grade carcinoma. The patient was subjected to an orchiectomy and hormone therapy (flutamide). Jaundice subsided and he is well after 3 years of follow up and maintained hormone therapy. PMID- 10835759 TI - [Pulmonary hypertension and HIV infection: report of a case]. AB - Pulmonary hypertension associated to HIV infection has been reported in the literature with increased frequency. Apparently, this condition has a faster clinical evolution and a higher mortality than primary pulmonary hypertension. The pathogenic mechanisms of HIV associated pulmonary hypertension and the influence of its treatment on patient's evolution are not well known. We report a 32 years old homosexual male that developed a severe dyspnea in a period of 2 months. Echocardiogram demonstrated right ventricular dilatation and a systolic pulmonary artery pressure of 86 mm Hg. No other causes for pulmonary hypertension were found. Antiviral therapy and vasodilator treatment with a calcium channel blocker were started and the patient had an important subjective clinical improvement. PMID- 10835760 TI - [Inactivation of tumor suppressor genes in uterine cervix carcinogenesis]. AB - The importance of inactivation of tumor suppressor genes in the development/progression of carcinomas of the uterine cervix is reviewed. It is well known that HPV-related oncogenes are strongly linked to cervical cancer. However, fewer studies have explored the occurrence of inactivation of tumor suppressor genes in this neoplasia. Genetic deletions affecting tumor suppressor genes are the most common mechanism of inactivation of these genes. Studies using conventional molecular techniques such as restriction fragment length polymorphism (RFLP) and Southern Blot showed low frequency of deletions in cervical carcinomas. Detection of deletions by using RFLP and Southern Blot presents several disadvantages, the most important being the difficulty in analyzing pure tumor cells. More sensitive approaches include tissue microdissection and PCR analysis of micro-satellites. Using these approaches, it has been shown that genetic deletions are, in fact, frequent events in cervical cancers, being detected in up to 95% of the cases. Multiple genetic loci are involved, including chromosomes 3p, 5p, 6p and 11q. Deletions are detected even in precursor lesions (cervical intraepithelial neoplasia, CIN). Some deletions have been correlated with prognostic parameters, such as stage, depth of invasion, and vascular space involvement. It is concluded that cervical carcinogenesis, like in other tumors, is a multistep process, characterized by the accumulation of events including activation of oncogenes, as well as inactivation of tumor suppressor genes. PMID- 10835762 TI - [The history of genetics in Latin American countries during the twentieth century]. AB - The aim of this historical review is to evaluate the evolution of genetics in the context of Latin American scientific culture, to value foreign influences and to highlight the discoveries and contributions of Latin American geneticists. During the first third of the twentieth century, local naturalists, botanists and physicians understood the chromosomal theory of heredity and Mendelian theory of evolution and begun research and teaching on these new theories and technologies. During the thirties and forties, North American geneticists visited South America and formed development poles on cytogenetics and population genetics in Brazil and Argentina. During the fifties and sixties, human genetics was formally established in Brazil, Argentina and Chile. Genetics teaching became generalized in universities and national Genetics Societies were formed. In 1969, the Latin American Genetics Society was created, unifying the efforts of zoologists, botanists, physicians and anthropologists in an unique Latin American cultural space, organizing 11 meetings between 1972 and 1994. Latin Americans have made a great contribution in genome discovery of animal, vegetable and human species in their territory. They explored the great genetic diversity of the continent, discovering new genes and diseases. The biomedical area had the greatest development. In 1997, there were 130 medical genetics centers, 120 hospitals specialized in congenital malformations, 56 molecular biology centers and 26 molecular genetics centers. At the end of the twentieth century, human genetics is completely integrated to medical sciences in Latin America. PMID- 10835761 TI - [Hantavirus in Chile: review and cases analysis since 1975]. AB - Since the identification of the first case of Hantavirus infection in Chile in 1995, a great advance in the knowledge of the disease natural history has occurred. There is evidence to suppose that the infection by Andes virus in suburban areas of Argentina and Chile, whose natural reservoir is Oligoryzomys longicaudatus, has been present but unidentified for a long time. We describe a serologically proven case occurred in 1975. The clinical presentation of the disease slightly differs from that described for Hantavirus Pulmonary Syndrome (HPS), caused by Sin Nombre virus in the USA. There is a wider range of presentation forms including, besides HPS, mild and asymptomatic cases, a greater proportion of renal involvement and pediatric cases. Therefore the epidemiological scenario would be an endemic disease with fluctuations that follow changes in rodent population and their interactions with humans. PMID- 10835763 TI - [Dobutamine and amrinone in septic shock patients]. PMID- 10835764 TI - Designing a bridge for consciousness: are criteria for a unification of approaches feasible? AB - Different intrinsic rules of functioning for body and mind are overemphasized by the mutually exclusive methods of investigation in both consciousness and the sciences. Classical science, alternative and complementary healing, quantum physics, and metaphysics all take drastically different approaches to consciousness research. A bridge is needed to restore the conceptual unity of mind-body. However, the construction of such a bridge paradoxically must rely on consciousness for information. The construction company (Consciousness, Inc.) therefore becomes means and end in the design of the bridge. This piece argues that the methods of each discipline contain research tools and resources to create a bridge that satisfies a unified multidimensional and multidisciplinary conception. The piece further suggests that the framework of the bridge must place consciousness as a partner to physiology in the evolutionary trajectory of life, since problems of survival are a primary consideration in the activity of both. Finally, the piece develops six criteria by which to evaluate the effectiveness of any construction design. PMID- 10835765 TI - Consciousness as cooperation. PMID- 10835766 TI - Living energy systems, and the "bridge problem". PMID- 10835767 TI - The search for an integral theory of consciousness. AB - In summary, Miller has identified one of the key concerns of consciousness research. However, significant contributions to an integral theory of consciousness will need to be more clear and precise and to incorporate the relevant literature on current integral theories. If Miller's future writings do this, they may make the contributions we so desperately need. PMID- 10835768 TI - A new study (Watson et al.) on "fighting spirit" and breast cancer. PMID- 10835769 TI - Beyond fighting spirit: what mind-states influence cancer survival? PMID- 10835770 TI - Intentional systemic mindfulness: an integrative model for self-regulation and health. AB - Self-regulation is the process whereby systems maintain stability of functioning and adaptability to change. Self-regulation is based on feedback loops which can be enhanced through attention. All self-regulation techniques, therefore, involve the cultivation of attention. However, the intention with which attention is directed may be crucial. In this paper, we explore intentional systemic mindfulness a model that explicitly introduces intention into self-regulation theory and practice. Intention as defined by this model is composed of the context of attention-systemic perspectives - and the quality of attention - mindfulness qualities. Intentional systemic mindfulness addresses both "why" (systemic perspectives) and "how" (mindfulness qualities) one directs attention, which may promote healing on multiple levels. Directions for research and implications for multiple levels of integrative health are considered. PMID- 10835771 TI - Disclosure and autonomic autopoiesis: a research and treatment model for twenty first century cancer survivorship. PMID- 10835772 TI - Detachment, hope, and spiritual understanding: a comment on Bernie S. Siegel's prescriptions for living. PMID- 10835773 TI - GPs caring for populations. PMID- 10835774 TI - Telehealth. What does it offer for public health care? PMID- 10835775 TI - Sexual abuse and personality disorders. PMID- 10835776 TI - Breast cancer and COCP. PMID- 10835777 TI - Is suffering part of being human or are good times really bad times? PMID- 10835778 TI - Controlling angina. Management options. AB - BACKGROUND: Angina pectoris is a common medical problem in the community requiring a multifaceted approach to treatment in order to improve the quality of life and long term prognosis for the patient. OBJECTIVE: To discuss some of the practical aspects of the medical management of patients with stable angina pectoris, including anti ischaemic medications and secondary prevention strategies. DISCUSSION: Decreasing the risk of rapid progression of atherosclerosis in patients with angina is probably the most important factor in long term management. Also, careful use of anti ischaemic and anti platelet medications may prevent further serious cardiac events and improve quality of life. PMID- 10835779 TI - Chronic heart failure. A guide for practical management. AB - BACKGROUND: Chronic heart failure is an increasingly common problem in Australia and most management occurs in the outpatient setting. OBJECTIVE: To review the options available in the management of chronic heart failure. DISCUSSION: Appropriate management can dramatically improve the prognosis, symptoms and quality of life of chronic heart failure patients. PMID- 10835780 TI - Common congenital heart defects. The value of early detection. AB - BACKGROUND: Congenital heart disease forms the largest group of congenital anomalies and therapy is available for all conditions. The main challenge is the early detection and referral of such cases. OBJECTIVE: The important features of the commonest congenital heart defects are reviewed with an emphasis on the diagnostic features. The pivotal role of the general practitioner in the early detection and referral of these patients is emphasised. DISCUSSION: A high index of suspicion, general understanding of transitional changes, haemodynamic aspects, good listening skills and examination techniques with the appropriate use of investigative tools, especially echocardiography will enable the practitioner to have a higher success with early detection and screening of patients. Rapport with institutions and parents is of great importance. PMID- 10835781 TI - The management of acute myocardial infarction. Does thrombolysis have a place? AB - BACKGROUND: Myocardial infarction is a common life threatening medical emergency. The mortality remains around 10% and can be influenced by early reperfusion achieved either by thrombolysis or angioplasty. OBJECTIVE: To outline the relative suitability and safety of thrombolysis as a treatment option for myocardial infarction. DISCUSSION: Initial assessment and treatment are vitally important in improving outcome. Later risk factor modification remains an important factor in reducing future cardiac events. PMID- 10835782 TI - The new antipsychotics. How much better are they? AB - BACKGROUND: Despite considerable efficacy, conventional antipsychotics cause debilitating extrapyramidal side effects and may worsen 'negative' symptoms of schizophrenia. New atypical antipsychotics (clozapine, risperidone, olanzapine) have become available, and quetiepine is expected shortly; it is TGA approved and is currently available to psychiatrists through a familiarization program. OBJECTIVE: To inform general practitioners about the novel antipsychotics and their therapeutic role in contrast to conventional antipsychotic drugs. DISCUSSION: The major benefit offered by atypical antipsychotics is that these drugs are less likely to cause distressing and disabling extrapyramidal side effects at therapeutically effective doses than conventional drugs. Advantages in terms of efficacy are marginal, though clozapine is superior to conventional drugs in treatment resistant patients. The new drugs are not without other side effects. Clozapine can cause agranulocytosis, needs ongoing haematological monitoring and is only available through specialist centres. However, the new drugs do improve overall outcome. Use of risperidone, olanzapine and quetiepine is now widely recommended in preference to conventional drugs, while clozapine is used when other drugs have failed. PMID- 10835783 TI - Breathlessness and pregnancy. AB - BACKGROUND: Breathlessness is common during pregnancy and is usually due to hormonal changes or mechanical factors. However, it is important to ensure that no other cause for this symptom is present, and to exclude underlying or co existent conditions. PMID- 10835784 TI - Locally advanced basal cell carcinoma. Is a cure possible? PMID- 10835785 TI - Cardiology quiz. Is the ECG always useful to diagnose ischaemia? PMID- 10835786 TI - Pathology quiz. A pigmented skin lesion. AB - Melanomas can usually be distinguished histologically from benign lesions by the application of accepted criteria of high discriminatory value. Occasionally, the distinction cannot be made with certainty. It is convenient to consider cutaneous melanoma as essentially of one type, with the prognosis being mainly determined by the thickness of the lesion. There is a trend to narrower excision margins with adequacy being judged solely by careful histological assessment. A formula driven approach should be avoided. PMID- 10835787 TI - Radiology quiz. Altered bowel habit. PMID- 10835788 TI - Enhanced primary care package. PMID- 10835789 TI - Could repeat prescriptions identify patients needing extended medical review? An exploratory study. AB - INTRODUCTION: New Medicare Benefit Schedule care planning items require general practitioners (GPs) to identify their patients with chronic medical conditions and multidisciplinary care needs. This paper investigates the use of long term repeat prescriptions to identify general practice patients who meet these criteria. METHODS: Twenty-nine GPs in urban, rural and inner city locations recruited 10 consecutive patients on repeat prescriptions for 12 months for medical (not primarily psychiatric) conditions. These patients reported their disease, service and morbidity (SF-36) profiles. GP attender profiles were compared with chronic disease self help group members (n = 44), an ACT community sample (n = 555) and a national GP consultation survey sample (n = 31,575). RESULTS: GP patients reported common chronic disease profiles, high levels of specialist, hospital and medication usage, with infrequent use of allied health professionals and self help groups. They reported considerable physical and psychosocial morbidity comparable to or worse than other groups. DISCUSSION: GP patients requesting repeat prescriptions may have substantial physical, psychological and social needs that might be met using a multidisciplinary team approach with allied health professionals and self help groups. CONCLUSION: GPs can use long term repeat prescriptions to identify a chronic disease cohort who could benefit from health care planning under the new Medicare Benefit Schedule funding arrangements. PMID- 10835790 TI - Realities of practice. Engaging parents and GPs in developing clinical practice guidelines. AB - OBJECTIVE: To integrate evidence based medicine with the experience and expectations of consumers and GPs in the development of clinical practice guidelines for acute respiratory infections (ARI) in young children. METHOD: Focus groups and workshops were held with 21 GPs and 27 patients of young children involved in a 2 year randomised controlled trial. RESULTS: The acceptability of the guideline development process for participants was determined. Barriers were identified which would impede clinical change, including: inadequate time; lack of knowledge; fear of patient dissatisfaction; and fear of poor health outcome. CONCLUSION: This paper details a process of guideline development that addresses the realities of general practice in Australia and the concerns of consumers. We identified potential barriers to change and integrated intervention strategies with the evidence to produce realistic clinical practice guidelines. PMID- 10835791 TI - Geriatric case managers: integration into physician practices. AB - Integration of case management proved to be a key variable in a national demonstration at nine sites of alternative models designed to enhance primary care of frail elders. Numerous semistructured interviews of participants revealed a number of areas important to achieving successful integration. These areas include making favorable first impressions, building relationships, learning to collaborate, having proximity and contact, communicating, and demonstrating benefits to patients and physicians. PMID- 10835792 TI - Case management within substance abuse treatment programs in Los Angeles County. AB - Case management has been used to link clients and the service system, but is loosely defined and poorly understood. The aims of this study were to describe the use and purposes of case management within substance abuse treatment programs, and also the structures and processes for providing case management services. A descriptive survey was done, with 50 program directors of 134 treatment programs (with 205 case managers), in Los Angeles County. Results showed that 80% of directors reported they use case management. Half of the programs use case management both before and after discharge, and 72% provide case management to all clients. Case managers' most important roles are to develop treatment plans and prevent relapse during treatment, and 60% of directors indicate the case managers in their programs are also counselors. Case managers perform numerous roles of coordination and advocacy. Eighty percent of case managers follow the care of the client during treatment and 32% of directors reported a case load of 1-10, while 26% reported a case load of 11-20. The professional background of case managers varies from chemical dependence professional to social worker or nurse. Twenty percent of program directors plan to increase case management in the future. PMID- 10835793 TI - HIV/AIDS case management: views from the frontline. AB - Case management has been a critical component of services to vulnerable populations for the past 20 years, and the knowledge base has been constantly evolving. This article offers an additional dimension to the study of case management practice by providing an opportunity for perceptions of case managers, heretofore relatively neglected, to be included in the literature. It reports the results of two study efforts (chart review and focus groups) that examined the case management system of care for people with HIV/AIDS in the New York tri county region. While the literature and chart review findings stress the primacy of linkage activities, the case managers emphasize the importance of providing the support necessary to ensure that clients are able to maintain a reasonable quality of life. Findings are discussed in the context of the "strengths perspective" as well as service system and organizational contingencies. Potential methodological implications for the use of the study techniques are suggested. PMID- 10835794 TI - Case manager-defined roles in the Medicare Alzheimer's Disease Demonstration: relationship to client and caregiver outcomes. AB - This study explores the different approaches to case management within, the Medicare Alzheimer's Disease Demonstration. Eight sites from around the country were selected to participate in a survey of case managers (N = 57) that evaluated their professional background and experience and how certain tasks, functions, and goals of case management were prioritized at each site. Client and caregiver outcomes were collected on a site-by-site basis as part of the demonstration. The hypothesis was that the way in which case managers performed their work would vary by site. Furthermore it was hypothesized that these differences would be measurable and correlated with project outcomes. Case managers differed significantly by site in how they prioritized tasks, functions, and goals, viewing themselves along a continuum from a clinical approach to one that emphasizes service management. These differences in case management style are reflected in between-site differences in certain client and caregiver outcomes but not others. Specifically, variations in case management style are not related to client cognitive or functional status but to other outcomes including behavioral management, caregiver burden, and service use. PMID- 10835795 TI - Living in limbo: when a spouse or close family member has an incapacitating illness. PMID- 10835796 TI - Elderdesign: home modifications for enhanced safety and self-care. PMID- 10835797 TI - Research challenges to recruitment and retention in a study of homebound older adults: lessons learned from the nutritional and dental screening program. AB - This paper attempts to describe the many reasons why it is difficult to recruit homebound older adults to participate in research studies. The reasons, not surprisingly, are more practical than theoretical. Complicated procedures for participants to follow, cross-cultural communication, fear of the medical establishment, convoluted consent forms, making contact on the telephone, finding the participants' homes, and a host of other nagging problems combine to erect enormous barriers to running successful studies. It is our hope that in identifying these difficulties we will also shed light on how to design studies that minimize their impact. PMID- 10835798 TI - The PACE model: description and impressions of a capitated model of long-term care for the elderly. AB - The Program of All-Inclusive Care for the Elderly (PACE) is an innovative model of comprehensive long-term medical and social services. The system receives monthly capitation payments from Medicare and Medicaid for clients who are nursing-home eligible. PACE was developed originally by On Lok Senior Health Services in San Francisco's Chinatown in the late 1970s. PACE is based upon the concept of maintaining the independence, autonomy and dignity of frail and disabled elders nearing the end of life (Sapir, 1996). The program's underlying principle is to keep participants (PACE program enrollees) in the community for as long as it remains medically, socially, and economically feasible (Shen and Iverson, 1992). There is also a strong intention to preserve and support the older adult's family unit. PMID- 10835799 TI - Music therapy in mobility training with the elderly: a review of current research. PMID- 10835800 TI - Regulation and functions of c-Jun N-terminal kinase/stress-activated protein kinase. AB - The cell uses a complex signal transduction network both to respond to changes in its microenvironment and to integrate all of its intracellular signals. Actions of various signals in the cellular signal transduction network converge at the mitogen-activated protein kinase (MAPK) cascades, which lead to and activate transcription factors in the nucleus and other effectors throughout the cell. Extracellular signal-regulated kinases (ERKs), c-Jun N-terminal kinases/stress activated protein kinases (JNK/SAPK), and p38 are some of the better characterized members of the MAPK family. Unlike ERK mainly responding to mitogens, JNK/SAPK was originally identified as a stress-related kinase and it has been extensively demonstrated to be associated with apoptosis. However, recent studies have further indicated that JNK/SAPK also has many functions other than the promotion of apoptosis. The repertoire of the JNK/SAPK pathway is wide ranging, and its functions can vary, and even oppose each other, depending on the cell types and stimuli. It remains a challenge for biomedical scientists to elucidate the distinct role of JNK/SAPK in each model system. PMID- 10835801 TI - The effect of incentive spirometry on chest expansion and breathing work in patients with chronic obstructive airway diseases: comparison of two methods. AB - BACKGROUND: Chronic obstructive airway diseases (COAD), characterized by mucus hypersecretion, lead to exercise intolerance. Incentive spirometry has been used to prevent postoperative pulmonary atelectasis. METHODS: To compare the efficacy of two incentive spirometers, Coach (volume-oriented) and Triflo (flow-oriented), in the work of breathing in COAD patients, 22 patients were randomized in this study: 12 patients (Triflo-II group) initially used Triflo-II for 10 minutes and then Coach for the same period. In contrast, the Coach group, including 10 patients, started with Coach followed by Triflo-II. After receiving incentive spirometry, lung expansion and work of breathing were assessed. RESULTS: Patients in the Coach group significantly increased chest wall expansion (p = 0.041), as compared with patients using Triflo-II. Similarly, there was also a significantly increased abdominal wall expansion in the Coach group (p = 0.0056), compared with that in the Triflo-II group. The need of accessory muscle assistance for breathing in the Coach group was significantly less than in the Triflo-II group (p = 0.047). It was easier for patients in the Coach group to start a breath (p = 0.0058) than for those in the Triflo-II group. For the entire group, 17 patients (77.3%) preferred Coach to assist their breathing, and only 4 patients (18.2%) favored Triflo-II. CONCLUSION: COAD patients achieved a larger expansion of the chest and abdomen with a Coach device. Our data provide a good rationale for an outcome study on the use of incentive spirometer in COAD patients. PMID- 10835802 TI - Experience in the treatment of myasthenia gravis with double filtration plasmapheresis. AB - BACKGROUND: Myasthenia gravis (MG) is an autoimmune disease characterized by production of autoantibodies directly against acetylcholine receptors on postjunctional membranes. The treatment modalities of myasthenia gravis include cholinesterase inhibitors, surgical thymectomy, immunosuppressive treatment, and short-term immunotherapies, including plasmapheresis and intravenous immune globulin. Double filtration plasmapheresis (DFP) is a new technique of plasmapheresis. The aim of our study is to evaluate the effectiveness and complications of DFP in the treatment of MG. METHODS: From December, 1993 to August, 1998, a total of 33 patients with MG received 68 courses (total of 299 sessions) of DFP in our hospital. Plasmapheresis volume, drainage volume, treatment duration, heparin dose, and complications were recorded. In addition, clinical responses after DFP were also evaluated. RESULTS: Only 4 courses (5.88%) of treatment were ineffective. Sixty-four courses (94.12%) of treatment lessened disease activity. The complications of DFP included 15 episodes of hypotension (5.01%), 2 of bradycardia (0.67%), 3 of chest pain (1.00%), 7 of dizziness (2.34%), 58 of high transmembranous pressure of plasma separator (19.40%), 3 of high secondary pressure of plasma fractionator (1.00%), 12 of hemolysis (4.01%), 2 of plasma separator clot (0.67%), 2 of plasma fractionator clot (0.67%), 3 of blood leakage (1.00%), and 1 of air embolism (0.33%). There was no mortality associated with the DFP procedure. CONCLUSION: DFP is a safe, convenient, and time-saving therapy with rare severe complications in the treatment of MG. Our experience confirms its effectiveness when used in combination with drug therapy and thymectomy. PMID- 10835803 TI - Laparoscopic extraperitoneal sacrospinous suspension for vaginal vault prolapse. AB - BACKGROUND: A new laparoscopic sacrospinous suspension procedure is described for the correction of vaginal vault prolapse using an extraperitoneal approach. This is the first report in the literature of the extraperitoneal approach. METHODS: We reviewed 12 women who had been treated in our hospital using this technique because of vaginal vault prolapse. These women had undergone hysterectomies (10 abdominal; 2 vaginal) between 5 and 22 years previously (mean, 12 years). After pre-laparoscopic preparation, a 10-mm trocar with a 10-mm zero-degree telescope was placed into the Retzius space. Using a direct air-distended method with a 20 mmHg insufflation pressure, Retzius and para-rectal spaces were created. The sacrospinous ligament could be easily identified and confirmed. A permanent suture was then inserted from the sacrospinous ligament to the vaginal vault to ensure that there was no space in between. RESULTS: This procedure was followed for all 12 patients. There were no major complications during surgery. Eleven women had no recurrence of vaginal vault prolapse during a follow-up period of 1 to 3 years (mean, 2.2 years). One patient developed recurrent vaginal vault prolapse; however, she subsequently underwent a successful colposacropexy by laparoscopy 23 months after the initial surgery. CONCLUSION: We modified the traditional sacrospinous fixation laparoscopically, following principles to restore the correct anatomic position of the vault. Laparoscopic extraperitoneal sacrospinous suspension can eliminate the procedure of opening and closing the peritoneum and avoid interference with the intestine during surgery. It can be used as an alternative to traditional genital suspension surgeries. PMID- 10835804 TI - A physiological method for the repair of young adult simple isthmic lumbar spondylolysis. AB - BACKGROUND: Anterior or posterolateral spondylodesis has been reported as a widely used surgical treatment for lumbar spondylolysis and spondylolisthesis. However, the expenditure of energy for both the physician and the patient is relatively great. The risk of complications is significant and the loss of one motion segment must be accepted. The latter is problematical, especially in young adult patients who only have symptomatic isthmic lumbar spondylolysis. METHODS: For these patients, direct repair of the defect is recommended. We used a spondylolysis hook screw and bone grafting for direct repair of the pars defect. The results were analyzed according to the clinical results and functional x-ray exams. RESULTS: The clinical results were good or excellent. The roentgenologic examinations at 6 months showed the presence of bone trabeculation across the pars defect in all cases. The physiological motion of the lumbar spine was preserved without limitation, and all of the patients had resumed daily activities or work without discomfort. CONCLUSION: There were no complications among our cases. Particularly, according to our anatomical investigations, normal structures were safe from injury as long as the proper technique was followed. A spondylolysis hook screw with bone grafting can preserve the physiological motion of the offending levels. This method is a much more physiologically appropriate surgical method for non-complicated and symptomatic lumbar spondylolysis in the young adult. PMID- 10835805 TI - Lethal midline granuloma: report of three cases. AB - Lethal midline granuloma (LMG) is a rare clinical entity characterized by progressive relentless ulcerations and necrosis of midfacial structures. It occurs more frequently in Oriental than in Western populations with no demonstrable etiology. Treatment and outcome for cases differ, but their pathological distinction may not always be possible from routine biopsy specimens. The histological features often seen include widespread coagulative necrosis, heavy inflammatory infiltrates, and atypical pleomorphic cells. However, the paucity of these atypical cells in biopsy specimens and the degree of necrosis can make the diagnosis of a neoplastic lesion very difficult. Because of the progress in pathology methodology including immunohistochemistry, most cases have been proven to be malignant lymphomas of T-cell lineage. We present 3 patients for whom an initial clinical diagnosis of LMG was made. From their several oral biopsies and nasal specimens, difficulties were encountered in differentiating "midline granuloma" from other possible diseases using histomorphological criteria alone. After extensive evaluations, malignant T-cell lymphoma was the specific disease entity identified in only one case by cell membrane immunostaining technique. A literature review was carried out, and recent concepts of the etiology and pathogenesis of this disease are presented. PMID- 10835806 TI - Reconstruction of upper chest wall defects with a function-preserving pectoralis major muscle flap: case report. AB - The pectoralis major muscle or myocutaneous flap has a nearly 100% success rate in reconstructing chest wall defects. Major adverse sequelae resulting from the use of the pectoralis major muscle or myocutaneous flap are rarely reported in the literature. However, the loss of pectoralis major muscle function caused by the detachment of the muscle from its insertion on the humeral bone is of more and more concern. This is a significant loss for manual laborers when the patient tries to handle tools or control heavy machinery. A case of upper sternal osteomyelitis is reported. After wide debridement with partial excision of the sternum, the second and third ribs, the right pleura, and the lung were exposed. A right unilateral pectoralis major muscle flap was transposed to restore the defect. In addition, to preserve the lateral portion of the muscle and its insertion on the humerus, the origin of the lower sternocostal part of the pectoralis major muscle was transposed to the medial clavicle and residual upper sternum. In this way, not only was the chest wall defect reconstructed but the function of the residual pectoralis major muscle was also preserved. Postoperative follow-up at one year demonstrated no arm weakness, no limitation in shoulder range of motion, and no evidence of atrophy of the transposed pectoralis major muscle. Our experience with this function-preserving pectoralis major muscle flap was encouraging and we suggest it be employed in the reconstruction of the upper anterior chest wall. PMID- 10835807 TI - Recurrent lacrimal sac papilloma: case report. AB - Tumors of the lacrimal sac are rare. Benign papillomas comprise approximately 40% of all neoplasms of the lacrimal drainage system. They often present insidiously with symptoms of dacryostenosis or dacryocystitis. Recurrent bouts of dacryocystitis and nasolacrimal duct obstruction were reported in a 35-year-old man over a period of 13 years. A medial canthal mass was noted in the 6th year after the onset of symptoms. A tumor was discovered incidentally during surgical intervention for presumed dacryostenosis. Surgical removal of the tumor and dacryocystorhinostomy were performed. The histopathologic report turned out to be benign papiloma. Local recurrences occurred during the follow-up period. In addition to surgical excision, we applied cryotherapy and CO2 laser to prevent further recurrence. This case we presented the characteristic recurrence of lacrimal sac papilloma and implied the possibility of tumor occurrence in a patient with recurrent dacryocystitis. We must bear in mind that a recurrent dacryocystitis may be a presentation of a lacrimal sac tumor, because early diagnosis and aggressive treatment can prevent recurrence and result in a cure. PMID- 10835808 TI - Nasopharyngeal carcinoma with leptomeningeal dissemination: case report. AB - Nasopharyngeal carcinoma (NPC) is a highly prevalent malignancy in southeast China, Hong Kong, and Taiwan. Spread of this tumor is known to occur via three main routes, i.e., local invasion of adjacent structures, regional metastasis to neck nodes, and hematogenous metastasis to distant organs. In this report, we describe a rare case of NPC disseminated via the leptomeninges, so called meningeal carcinomatosis (MC). The patient was a 62 year-old man who presented with multiple cranial nerve palsies and a headache, and was diagnosed with NPC in August 1988. The primary tumor regressed completely after induction chemotherapy and radiation therapy. Computerized tomography (CT) 17 months after radiation therapy showed multiple enhanced nodules scattered along the temporal meninges. The nodules increased in number and size in the subsequent CT scan 4 months later. The patient declined further invasive procedures and oncologic treatments, and he expired at home 9 months after the development of MC. It is speculated that perineural invasion and access to the subarachnoid space was the major cause of MC in this case. The case, although rare, possibly highlights a rare route of tumor dissemination in NPC. PMID- 10835809 TI - Stand-alone database for an air transport program. AB - This case study demonstrates both the positive and the negative aspects of developing a stand-alone database for use by a small unit of a larger organization. The advantages center on the ability to tailor the program to the requirements of a single unit. The disadvantages relate to the inability to apply systemwide resources to the problem, the dependence upon a small group of experts who are able to modify and maintain the program, and the inability to share data across organizational units. Units that are contemplating the development of an isolated database to meet the needs of a small unit are advised to consider their development in light of the needs of the entire organization. A unit should explore all other options for meeting its and information management needs before embarking on an isolated database project. They also should identify a method for eventual consolidation of information resources. Finally, when at all possible, a solution should not center upon the talents of a single individual. This approach sets up the organization for problems when that individual leaves the organization or moves on to another position. PMID- 10835810 TI - Development and test of a model for designing interactive CD-ROMs for teaching nursing skills. AB - The use of interactive multimedia is well documented in the education literature as a medium for learning. Many schools of nursing and healthcare agencies purchase commercially-made CD-ROM products, and, in other cases, educators develop their own. Since nurses are increasingly designing CD-ROMs, they must be aware of the instructional design needed to develop comprehensive and effective CD-ROMs that do not compromise the quality of education. This article describes a process for developing and testing an interactive, multimedia CD-ROM on oral medication administration, using an instructional design model based on Chickering and Gamson's Principles of Good Practices in Education. Results from testing the model are reported. The findings can be used to guide the work of nurse educators who are interested in developing educational software. PMID- 10835811 TI - Creating Web-based, multimedia, and interactive courses for distance learning. AB - A case study describes how faculty at Saint Louis University School of Nursing have developed computer-based, multimedia courses for master's and post-master's nursing education. Lectures with slide presentations are recorded digitally and encoded for multimedia streaming over the World Wide Web for distance learning. The technology is explained in detail in terms of the specific technologies used for lecture and course development. How this technology supports graduate student learning is also presented. PMID- 10835812 TI - Predictive accuracy of the HESI Exit Exam. A follow-up study. AB - This follow-up study compares the accuracy of the HESI Exit Exam (E2) in predicting NCLEX success for the academic years 1996-97 (N = 2,809) and 1997-98 (N = 3,752) and is designed to replicate the study of Lauchner et al. and to implement recommendations suggested by the authors for further research. The E2 was again found to be highly predictive of licensure success, regardless of the type of program tested: associate degree, baccalaureate degree, diploma, or practical nurse programs. The predictive accuracy of the E2 was 98.27% for the RN group and 99.34% for the PN group. The E2 was again found to be significantly more accurate when administration of the exam was monitored than when it was not monitored (P = 0.05). In the 1997-98 academic year, NCLEX success of low-scoring E2 students was examined. Significantly more (P = 0.001) of the low-scoring E2 students failed the licensure exam than high-scoring E2 students. However, significantly fewer (p = 0.05) of these low-scoring E2 students failed the licensing exam when the E2 was used as a benchmark or guide for remediation. PMID- 10835813 TI - Bringing decision support to nurse managers. AB - Modern hospital nursing management requires timely and accurate information to allow nurse managers to adjust resources to patient requirements. We report an experience using production theory to provide the framework for the development of Decision Support Objects: graphic displays of nursing hours budgeted, scheduled, and worked within institution-specific control limits. Every month, nurse managers follow an analytic algorithm to understand nursing resources and trends on their units. Exception reporting closes the accountability loop. The essentials of education for nurse managers include skill training in the use of the decision-support tools and supportive lecture/seminars for understanding the managerial implications of using them. PMID- 10835814 TI - The respiratory health effects of passive smoking on children. PMID- 10835815 TI - Asthma treatment and noncompliance. AB - RATIONALE: Noncompliance with asthma treatments is a common cause of hospitalization. Currently, physicians in the emergency department can only diagnose noncompliance with asthma therapy by asking patients if they have taken their medication (i.e., the HONOR SYSTEM). Patients frequently "overstate" their compliance and subsequently receive unnecessary supplemental medication (p.o. corticosteroid) as well as hospitalization. However, because their noncompliant behavior goes unrecognized, these individuals are at risk for recurrent emergency care and death. It is for these reasons that drug monitoring to identify compliance status should be available in the emergency department and in specialty clinics that care for chronic asthmatics. RECOMMENDATIONS FOR ACTION: 1) Develop drug monitoring systems and make them available. There are two electronic medication-monitoring devices with microprocessor technology available for metered-dose inhalers. These devices need further improvement in design, at a lower cost. Assays have been developed for oral and injectable medications recently approved by the FDA. These assays need to become commercially available. 2) Train health care providers to improve patient compliance. As noncompliance becomes more recognized, as a result of drug monitoring, educational and behavioral programs to improve compliance with treatment will need to be developed and disseminated. PMID- 10835816 TI - Creating a physician web site: Part One. AB - The reasons why physicians should have professional web sites for their practices are presented. The options available to practitioners who want to create a site are reviewed. Advantages and disadvantages of virtual and non-virtual hosting are discussed. How to find and register one's own domain name, obtain a hosting service, and avenues available to build the web site are also described. Future articles will address finding specific hosting services and details regarding site construction. PMID- 10835817 TI - What to look out for if offered a reimbursement schedule that's less than Medicare. PMID- 10835818 TI - Nature's way to good health. PMID- 10835819 TI - [The state of dentistry education in Europe]. AB - In order to find out what branches of Dentistry are recognized in Europe as Dental specialties a questionnaire was designed and sent to various Associations and groups across Europe. It contained eight main questions covering the various specialties that were recognised or were to be recognised. Responses were obtained from 28 countries, 15 EU member states and 13 non member states. The results from the questionnaire indicated that there was a broad agreement on many of the questions but a considerable variety of opinions. There seems to be a desire to harmonize standards across Europe for the benefit of the profession and the public. PMID- 10835820 TI - [Methodologic and systemic reasons for failures of metal ceramics]. AB - Only by the use of scanning electron microscope in investigations concerning the metal-porcelain interface of ceramic masses upon metal frames (approx. 160 crowns since 1967), essential differences in the reason of mistakes have been revealed recently. Primarily, methodic mistakes are to be declared relevant. Independent of their veneering origin, they contribute to fractures within the ceramic layer or to crack-offs of the fired ceramic masses from the metal frame. Obviously, the systemic causes for mistakes in metal ceramics revealed to be dependent on the groups of veneers. Veneering involves non-manageable reasons for mistakes. These qualities in the material are the coefficient of thermal expansion or the share of palladium in the veneer. Very often, palladium is responsible for the formation of bubbles in the ceramic layer, and thus it is the reason for the development of cracks. The so-called pure titanium is quite extraordinary, when covered with ceramic mass, because the so-called alpha case layer cannot be bridged, despite 10 years of permanent research in that field. The clinical critical examination of systemic mistakes, however, proves to be mild. Clinical reasons for mistakes in metal ceramics remain disregarded in this paper. PMID- 10835821 TI - [The use of the Dentatus articulator in complete denture prosthetics]. AB - The adjustable articulators and the bilaterally balanced occlusal scheme is not commonly used in Hungary, even if it is the mainly used type of occlusion in the world. The authors provide a short historical background about occlusal types then show how to use the Dentatus semi-adjustable articulator in the practice of the complete denture prosthetics: how to use the Dentatus ear-bow, and how to adjust individual values on the articulator. With the help of this procedures a bilaterally balanced occlusal scheme is designed, so the authors show the characteristics of this occlusal type. The aim of the authors is to present a view of "how to organize" occlusion for complete dentures. PMID- 10835822 TI - A new comprehensive study on aging--the National Institute for Longevity Sciences, Longitudinal Study of Aging (NILS-LSA). AB - A new comprehensive longitudinal study of aging, the National Institute for Longevity Sciences, Longitudinal Study of Aging (NILS-LSA) started in November 1997. The participants of this study will be 2,400 residents aged 40 to 79 years who were age- and gender-stratified random samples selected from the NILS area. All participants provided written informed consent after a detailed explanation of the study. They will be examined at the NILS-LSA Examination Center every two years. Their first wave examinations will be finished by the end of March 2000. The examined variables number over 1,000, including clinical evaluations, medical examinations, anthropometry, body composition, physical functions, physical activities, psychological assessments, nutritional analysis and molecular epidemiology. By the end of September 1999, 1,643 men and women had completed their first wave examinations. PMID- 10835823 TI - Ultrasonic evaluation of common carotid intima-media thickness (IMT)--influence of local plaque on the relationship between IMT and age. AB - Ultrasonic evaluation of the intima-media thickness in the common carotid artery (IMT-CCA) has been widely used as a marker of atherosclerosis. However, the definition of IMT-CCA is not uniform and it includes two quite different pathological changes; a general intima-media thickening and a local atherosclerotic change (plaque formation). The aim of this paper was to evaluate the IMT-CCA and local atherosclerosis separately, and to clarify how the IMT-CCA itself changes with age and how local plaques influence the relationship between the IMT-CCA and age. The subjects were 979 men and women aged 40 to 79 years who participated in the first wave examination of the National Institute for Longevity Sciences--Longitudinal Study of Aging (NILS-LSA). The IMT-CCA measured at the thinnest point was significantly higher in men (0.61 +/- 0.15 mm) than in women (0.58 +/- 0.14 mm, p < 0.01) and it increased with age (trend p < 0.0001) in both genders. The IMT-CCA was higher with the presence of plaque in the bulbs (PLQ-BLB) than without PLQ-BLB (p < 0.0001). Although this was a cross-sectional study, the IMT-CCA increased 0.06 mm/10 years with PLQ-BLB and 0.04 mm/10 years without PLQ-BLB. The IMT-CCA could be partially explained by age, gender and PLQ BLB (r2 = 0.317). In this middle-aged and elderly population, an increase in the IMT-CCA showed a moderate relationship with local atherosclerosis and age. PMID- 10835824 TI - Eye examinations at the National Institute for Longevity Sciences--Longitudinal Study of Aging: NILS-LSA. AB - The National Institute for Longevity Sciences--Longitudinal Study of Aging (the NILS-LSA) started in 1997, and involves many kinds of examination. The objective of this paper is to outline the eye examinations in the NILS-LSA. The eye examinations consist of checks on refractometry, visual acuity, intraocular pressure, contrast sensitivity, kinetic visual acuity, visual fields, fundus photography, and lens estimation. The subjects were 1,077 men and women aged 40 79 years who participated in the first year examination of the NILS-LSA. All subjective measurements (distant visual acuity, kinetic visual acuity, contrast sensitivity, and mean sensitivity of visual field) declined significantly from the 50s. Age-related structural changes in the lens or hypertensive and arteriosclerotic changes in retinal vessels began at least in the 40s. It is suspected that aging affects the subjective visual functions from the 50s. However, changes in the structure of eye may begin before the 40s. The data from the eye examinations of the NILS-LSA are useful to assess the aging effects on vision and to investigate the relationship between visual function and physical or psychosocial health problems among the elderly. PMID- 10835825 TI - The effects of age on hearing and middle ear function. AB - Audiometric test results from 933 community dwelling males and females were presented to obtain the norm for each generation after middle age. Three aspects were adopted in this analysis; the cross-sectional aging transition of pure tone thresholds, the comparison in the self-perceived hearing difficulty among generations and the evaluation of middle ear function using multifrequency tympanometry. Subjects were divided into four age groups; 40s, 50s, 60s and 70s. There were statistically significant differences in pure tone thresholds between almost any two age groups at each frequency in both genders, especially at higher frequencies. The pure tone thresholds were also discussed in regard to gender difference and laterality. A contradiction between self-perceived hearing difficulty and auditory accuracy was observed in the elderly generation. Analysis of tympanometric measurements was performed on static admittance at 226 Hz, tympanometric peak pressure at 226 Hz and resonance frequency of the middle ear. These variables did not show any systematic aging change. However, it was considered they represented the reference values of each generation. PMID- 10835826 TI - Age and gender differences in skin sensory threshold assessed by current perception in community-dwelling Japanese. AB - The purpose of this study was to investigate age and gender differences in current perception thresholds (CPT) in Japanese citizens. CPT values at frequencies of 2000, 250 and 5 Hz reflect different types of peripheral sensory nerve functions. Since there have been only a limited number of reports which investigated CPT values in community-dwelling people, little is known about variations with age and gender. The present study therefore concentrated on a large population of 1632 individuals (men; 818 mean age +/- standard deviation 59.4 +/- 10.9, women; 814, 59.4 +/- 11.1) in a Japanese community. Significant gender differences in CPT values at 250 and 5 Hz were observed. Multiple comparisons among 4 age groups (40s, 50s, 60s and 70s) showed age-related differences in CPT values at 2000 and 250 Hz in both genders. However, age and gender interactions with reference to CPT values appeared to be different between these latter two frequencies. At 5 Hz, only men showed age-related variations. These results indicated gender differences in fiber-specific aging changes. PMID- 10835827 TI - Bone mineral density obtained by peripheral quantitative computed tomography (pQCT) in middle-aged and elderly Japanese. AB - To clarify age-related changes in bone mineral density (BMD) by peripheral quantitative computed tomography (pQCT), 1,124 Japanese middle-aged and elderly community-dwelling people were examined. The BMD of the trabecular bone was assessed at the distal part of the radius (D50), and the BMD of the cortical bone was assessed at the diaphysis of the radius (P100). P100 during age 40 to 49 was significantly higher in females (1359.6 +/- 10.7 mg/cm3, mean +/- SE) than in males (1253.5 +/- 9.5 mg/cm3), while there was no difference in D50, 245.3 +/- 5.1 mg/cm3 in females and 293.0 +/- 5.5 mg/cm3 in males. Females and males aged 50 to 59 lost 8.09 +/- 2.08 (mean +/- SE) mg/cm3 and 3.80 +/- 1.77 mg/cm3 of D50 every year, respectively. As for P100, females lost 25.1 +/- 4.48 mg/cm3, and males lost 6.37 +/- 3.89 mg/cm3 every year. Because of these gender differences, both D50 and P100 were significantly higher in males than in females aged 50 and over. Assuming that the average BMD between ages 40 and 44 was the maximum bone mineral density (BMD max), the percentage change from the BMD max with age was examined. Females aged 60 to 69 whose BMD were under 70% of the BMD max made up 73.9% in D50 and 23.2% in P100. Only 21.1% of males aged 60 to 69 showed less than 70% of the BMD max in D50 and only 3.8% in P100. The percentage decrease in BMD by age was larger in D50 than in P100 in both males and females. The individual difference in BMD was larger in D50 than in P100. These results suggest that pQCT may be useful to independently assess aging effects on cortical and trabecular bone density. PMID- 10835829 TI - The factors related to age awareness among middle-aged and elderly Japanese. AB - The purpose of this article was to expand past research by examining relationships between age awareness and related factors in Japanese middle-aged and elderly people. The subjects were 1,129 participants (575 men and 554 women, aged 40 to 79 years) of the National Institute for Longevity Sciences- Longitudinal Study of Aging (NILS-LSA). They were examined with a questionnaire and interview. As a result, chronological age, self-rated health, and visual and hearing ability tend to be the related factors for awareness of aging. Comparing the demographic and physiological factors, however, major life events, difficulties and daily life experiences showed a stronger influence on age awareness. These results provide an interesting basis for the future understanding of adult development and the meaning of aging. PMID- 10835828 TI - Distribution of geriatric disease-related genotypes in the National Institute for Longevity Sciences, Longitudinal Study of Aging (NILS-LSA). AB - Phenotypes of various genes related to geriatric diseases and the aging process were assessed in the National Institute for Longevity Sciences, Longitudinal Study of Aging (NILS-LSA). The subjects were 1,297 participants in the NILS-LSA. They were community-living males and females aged 40 to 79 years who were randomly selected from the area of the NILS. Genotypic and allelic frequencies of genes in the subjects were analyzed. Age and gender differences in the distribution of genotypes were also tested. The genotypic frequencies were as follows: (1) Angiotensin I-converting enzyme (ACE) genotype was I/I 46.2%, I/D 38.3% and D/D 15.5%. (2) alpha 1-adrenoreceptor genotype was C/C 84.4%, C/T 12.7%, and T/T 3.0%. (3) Apolipoprotein E genotype was epsilon 2/epsilon 2 0%, epsilon 2/epsilon 3 7.9%, epsilon 3/epsilon 3 70.0%, epsilon 3/epsilon 4 20.8%, epsilon 2/epsilon 4 0%, and epsilon 4/epsilon 4 1.4%. (4) Cholecystokinin type-A receptor (CCKAR) nucleotide-81 (nt-81) genotype was A/A 59.1%, A/G 35.1%, and G/G 5.9%. The CCKAR nucleotide-128 genotype (nt-128) was G/G 74.3%, G/T 23.6%, and T/T 2.2%. The combination of nucleotide (nt-81, nt-128) was (A/A, G/G) 59.1%, (A/G, G/G) 14.1%, (G/G, G/G) 1.1%, (A/G, G/T) 21.0%, (G/G, G/T) 2.6%, and (G/G, T/T) 2.1%. There were no subjects with (A/A, G/T), (A/A, T/T) or (A/G, T/T) genotypic combinations. (5) beta 3-adrenoreceptor genotype was T/T 66.8%, T/A 28.5%, and A/A 4.7%. (6) Dihydrolipoamide succinyltransferase (DLST) nucleotide 19117 genotype was A/A 25.1%, A/G 49.7%, and G/G 25.1%. The DLST nucleotide 19183 genotype was C/C 55.8%, C/T 38.2%, and T/T 5.9%. The combination of nucleotide (nt19117, nt19183) was (A/A, C/C) 6.7%, (A/G, C/C) 24.1%, (G/G, C/C) 25.1%, (A/G, C/T) 25.6%, (A/A, T/T) 5.9%, and (A/A, C/T) 12.6%. There were no subjects with (A/G, T/T), (G/G, T/T) or (G/G, T/C) genotypic combinations. (7) Transforming growth factor-beta 1 genotype T/T 35.2%, T/C 44.6%, and C/C 20.2%. (8) The platelet-activating factor acetylhydrolase genotype was M/M 71.7%, M/m 27.2%, and m/m 1.2%. The mitochondria DNA 5178 genotype A was 42.1% and C was 57.9%. There were no significant gender or age differences in tested genotypic and allelic distribution except for the DLST and apolipoprotein E. Differences in the genotypic frequencies of distribution using the Hardy-Weinberg equilibrium were significant in the ACE and alpha 1-adrenoreceptor genotypes. PMID- 10835830 TI - Effects of social support and self-esteem on depressive symptoms in Japanese middle-aged and elderly people. AB - We examined the relationship among social support, self-esteem, and depression. The subjects were 1,116 Japanese community-dwelling adults aged between 40-79, who were the first wave participants of the National Institute for Longevity Sciences--Longitudinal Study of Aging (NILS-LSA). Exploratory and confirmatory factor analyses were performed on the Rosenberg's self-esteem scale that supported the superiority of the bi-dimensional structure of the scale marked by self-confidence and self-deprecation subscales. The subsequent causal analyses, using structural equation modeling, demonstrated that social support reduced depressed affect through an increase in self-confidence and a decrease in self deprecation. By contrast, social support did not show a direct effect on depressed affect. The findings suggest the importance of esteem-improving elements of social support in reducing depressive symptoms. PMID- 10835831 TI - Nutritional assessments of 3-day dietary records in National Institute for Longevity Sciences--Longitudinal Study of Aging (NILS-LSA). AB - Food and nutrient intake of NILS-LSA participants 40 to 79 years of age were assessed through 3-day weighed dietary records by gender and age. The results were as follows. The intake of fats and oils, meats and beverages tended to decrease, but fruits increased with age in both males and females. Regarding nutrient intake, energy, protein, fat, and cholesterol showed a decrease as individuals aged. The nutrient intake in the 70 to 79 yr group was significantly lower than other age groups of both genders. The total dietary fiber and vitamin C intake increased. Salt intake exceeded 10 g/day in every group. The percentage of energy from fat to total energy was higher than 25% in most age groups. The proportion of fatty acids was almost appropriate in all groups. PMID- 10835832 TI - Age-related changes in gait velocity and leg extension power in middle-aged and elderly people. AB - To prevent a decline in gait with age, it is necessary to investigate age-related changes in gait performance and detect related factors. The purpose of this study was to assess the association between gait ability and leg extension power among middle-aged and elderly people. Height, weight, maximum gait velocity (MGV) and leg extension power (LEP) were measured in 752 males and females who participated in the National Institute for Longevity Sciences, Longitudinal Study of Aging (NILS-LSA). Age-related changes in MGV and LEP and the association between MGV and LEP were assessed. There were significant decline trends in height, weight, MGV and LEP with advancing age (p < 0.001). MGV showed a significantly positive correlation with LEP (in males: r = 0.48 p < 0.001, in females: r = 0.47; p < 0.001). Subjects aged 60 yrs and over showed a significantly higher correlation than those under 60 yrs in males, but not in females, after adjustment for height and weight. Although the relationships between MGV and LEP were different by age and gender, LEP may be one of the important factors in maintaining gait ability. PMID- 10835833 TI - Comparison between the air displacement method and dual energy x-ray absorptiometry for estimation of body fat. AB - Air displacement plethysmography (ADP) is a method for the determining percent body fat (%BF) using the two-compartment model, in which the body is partitioned into body-fat mass and fat-free mass (FFM). Although this model assumes a constant density of FFM as 1.10 g/ml, its density may depend upon the bone mineral content (BMC) and total body water (TBW) which vary with age, gender, and race/ethnicity. This study compared %BF determined from ADP (ADP%BF) with %BF obtained by dual-energy x-ray absorptiometry (DXA%BF), and also investigated the effects of BMC, TBW, and other factors on its value. The subjects were 721 female and male Japanese aged 40 to 79 years. Body density was measured by ADP and %BF was calculated using Brozek et al's equation. BMC and body-fat volume were measured using DXA, and TBW was measured by multifrequency bioelectrical impedance. A series of anthropometric measurements was taken. Although ADP%BF was highly correlated with DXA%BF (female: r = 0.89, male: r = 0.90) (p < 0.001), ADP%BF differed significantly from DXA%BF (female: -1.30 +/- 0.14% (mean +/- s.e.m.), male: 1.22 +/- 0.13%) (p < 0.001). The difference in %BF (ADP%BF-DXA%BF) was negatively associated with BMC/FFM but not with TBW/FFM in both genders. The difference in %BF was also positively correlated with waist circumference. Considering previous studies, this result may be explained by the underestimation of DXA%BF, rather than by the overestimation of ADP%BF. In conclusion, ADP may be a useful method to measure %BF. However, BMC should be taken into consideration. Furthermore, DXA%BF may be underestimated in people with large waists. PMID- 10835834 TI - Frequencies and circumstances of falls in the National Institute for Longevity Sciences, Longitudinal Study of Aging (NILS-LSA). AB - Frequencies and circumstances of falls were assessed among 1030 middle-aged and elderly people who participated in the NILS-LSA (National Institute for Longevity Sciences, Longitudinal Study of Aging) from November, 1997 to March, 1999 and responded to the questionnaires. The variables analyzed in the present study were demography and history of falls in the past one year. Circumstances of falls, e.g. time, location, activities associated with falls, cause of falls and degree of injury due to falls were asked when the subjects experienced a fall. Fear of falling was also investigated in all subjects. The prevalence of falls was 12.9% in the middle-aged group (40-59 yr.) and 16.5% in the elderly group (60-79 yr.). The distribution of time, location, activity associated with falls, cause and injury due to falls corresponded with previous fall studies among community dwelling elderly people. The incidence of falls was extremely high during the daytime and outdoors. Falls occurred most frequently while walking. The majority of falls were due to extrinsic factors. About 40% of all falls caused no injury. As to the fear of falling, about 30% of the middle-aged subjects and about 60% of the elderly subjects reported that they were fearful. Our results suggested that fall accidents are not rare, even in middle-aged people. PMID- 10835835 TI - Mesenteric venous thrombosis: a renewed diagnostic and therapeutic challenge. PMID- 10835836 TI - Completed suicides and emergency psychiatric evaluations: the Louisville experience. AB - Suicide is one of the most serious outcomes of psychiatric illness, and the most extreme intervention (involuntary hospitalization) can be exercised if this event is likely. Despite this, the rate of suicide has remained fairly consistent at 1.1-1.4%. In an ongoing effort of identifying factors that can predict subsequent suicides, we retrospectively examined the records of individuals who completed suicides in Jefferson County, January 1997 through September 1998, and who were evaluated at the Emergency Psychiatric Service (EPS) at University of Louisville Hospital. Fifteen of the 132 (11.4%) subjects who completed suicide were evaluated at some point in time at EPS. Only 8 (6.1%) were seen within 60 days of the fatal event. This represents less than 0.1% of the total 9,469 patients seen at the EPS during this time period. No specific factors could be identified that predicted imminent suicide. Given the inaccuracy in being able to predict suicide, clinicians need to continue to be vigilant when assessing acutely distressed substance abusing or psychiatric patients. PMID- 10835837 TI - Associations in 425 patients having temporomandibular disorders. AB - OBJECTIVES: Chronic facial pain in general and temporomandibular disorders (TMD) in particular are among the most common conditions affecting mankind. The objective of this study was to retrospectively evaluate standardized data on patients having TMD in order to seek trends and cross-correlations. METHODS: The patients' records were reviewed for 425 consecutive patients having TMD, examined and treated in one clinic by one dentist. Demographic information and signs and symptoms of TMD were tabulated and the features were cross-correlated to find underlying trends. RESULTS: TMD primarily affects women, as more than 84% of those affected were female. The most common feature (found in 84% of subjects) was facial muscular pain in the vicinity of the TMJ. In 42%, this pain was bilateral. Unilateral pain was almost equally distributed on the right or left side. Headaches were reported in 78% of patients with TMD. "Popping" noises in the TMJ on opening (54%) or closing (31%) the mouth, or stiffness of the neck (33%) were the next most frequent associations. CONCLUSIONS: The demographics for the studied population of TMD patients closely agrees with previous studies reported in the literature. This study reinforces the fact that pain associated with TMD primarily affects women. PMID- 10835838 TI - [Clinical analysis of nursing home-acquired pneumonia in a community hospital]. AB - To clarify the characteristic features of nursing home-acquired pneumonia in our community hospital, we performed a clinical analysis of 86 patients with nursing home-acquired pneumonia. The patients were divided into young and elderly groups. In the young group cerebral palsy was the underlying disease. In the elderly group, it was cerebrovascular attack. Although there were no differences in ADL, the nutritional condition of the young group was comparatively good, the isolated microorganism consisted of mostly Mycoplasma pneumoniae and the prognosis was good. The elderly group where the nutritional condition was poor, the patients were detected by non-respiratory symptoms and risk factors such as obvious episodes of aspiration led us to be concerned about the risk factors for nursing home-acquired pneumonia. The microorganism isolated from the sputum of the elderly group was frequently a multi-drug resistant microorganism such as Methicillin-resistant Staphylococcus aureus (MRSA) and polymicrobial infection. Their prognosis was poor despite treatment with multiple antibiotics. In the comparative study between survivors and non-survivors in the elderly group, risk factors such as hypotension, consciousness disturbance, the extension of infiltration shadows, respiratory failure, multiple organ failure and metabolic acidosis were influenced for the prognosis, but the isolated microorganisms and the antimicrobial agents were not concerned. PMID- 10835839 TI - [A study of bacterial meningitis in Hokkaido between 1994 and 1998]. AB - We evaluated 82 children with bacterial meningitis in 78 institutions in Hokkaido, Japan between 1994 and 1998. The mean number of cases per year was 16.4 (range, 9 to 30). A male predominance occurred with 52 males and 30 females (1.7:1). Median age was 11 months with a range from 1 day to 11 years. Seventy eight (95%) of the total were less than 5 years old with 55 (67%) less than 2 years old and 45 (55%) less than 1-year old. Four deaths occurred, giving an overall case fatality rate of 4.9%. Sequelae were seen at discharge in 29%, predominantly sensorineural hearing loss, epilepsy and development delay of varying severity. The main causative organisms were Haemophilus influenzae in 49 patients, Streptococcus pneumoniae in 14, Streptococcus agalactiae in 10 and Escherichia coli in 5. The frequency of poor prognosis was 50% for patients with meningitis due to S. pneumoniae or S. agalactiae compared with 27% for patients with meningitis due to H. influenzae. PMID- 10835840 TI - [Increasing nalidixic-acid-resistance in Salmonella strains isolated from sporadic cases in Tokyo]. AB - A total of 118 nalidixic-acid (NA)-resistant Salmonella strains consisting of 68 domestic strains and 50 imported strains isolated during 1988-1998 in Tokyo were examined regarding their annual incidence, serovars, drug-resistance patterns, and minimum inhibitory concentrations(MIC) to fluoroquinolones (NFLX, OFLX, ENX, and CPFX). NA-resistant strains accounted for 1.3% of all Salmonella (5,302 strains) isolated from domestic cases, and 2.5% of all Salmonella (1,981 strains) isolated from imported cases. The incidence of NA-resistant strains has increased since 1995, and it has been remarkable in imported cases. The results of the serotyping showed that the NA-resistant strains were classified into 25 serovars, excluding untypable strains. Among those, S. Enteritidis (21 strains), S. Blockley (13 strains), S. Litchifield (13 strains), S. Typhimurium (13 strains), S. Hadar (9 strains), and S. Virchow (8 strains) were predominant. Drug resistance patterns of NA-resistant strains, including other drugs (CP, TC, SM, KM, ABPC, ST, FOM, and NFLX) tested varied among the 26 types. Among those, multidrug-resistant strains accounted for 61.9% (73 strains), and one strain among them was high-resistant to NFLX. MIC distribution of NA-resistant strains to fluoroquinolones showed that the ranges of all drugs were 4-128 times higher than NA-sensitive strains used for controls. PMID- 10835841 TI - [The clinical feature of febrile episodes]. AB - Institutionalized elderly patients are at risk of nosocomial infection because of their compromised status by aging. To clarify the relationship between fever and disease in elderly patients, we analyzed 1,105 febrile episodes, the etiology of which were already diagnosed, of 443 patients (136 men, 307 women). All patients who were 65 years of age or older and who had been admitted to the hospital for more than 7 days had fevers above 37.5 degrees C recorded. The etiologies of the 1,105 febrille episodes were respiratory tract infection in 381 (34.5%), urinary tract infection in 263 (23.8%), other diseases in 164 (14.8%) and in 297 (26.9%) unknown. The episodes were categorized into two groups by the degree of initial fever: group A, 559 episodes (50.6%) of 37.5-38.0 degrees C and group B, 546 episodes (49.4%) of above 38.0 degrees C. Of the episodes, 41.0% were one-day fevers, 21.4% two-day fevers, and 14.0% three-day fevers. The frequency of two-or more-day fevers was significantly higher in group B (69.0%) than in group A (49.2%) (p < 0.001). In group B, respiratory tract infection (44.3%) was more frequent than urinary tract infection (16.1%) (p < 0.001). Of the respiratory tract infections, 63.5% were in group B and, in contrast, 66.5% of the urinary tract infections were in group A. The white blood cell count and C-reacting protein levels were significantly higher in group B than in group A (p < 0.001). The degree of initial fever is an important predictive marker of severity of disease in elderly patients. PMID- 10835842 TI - [HIV-1 (human immunodeficiency virus type 1) mutant strain associated with zidovudine resistance detected from a Japanese patient with primary HIV-1 infection]. AB - Sequences of HIV-1 pol gene were determined by direct sequencing from a Japanese patient with primary HIV-1 infection. The patient did not receive antiretroviral therapies. However we observed a HIV-1 mutant strain associated with zidovudine (ZDV) resistance. The patient had both the codon 70 and the codon 215 amino acid substitutions in the RT region. Our data indicated that the patient was infected with a HIV-1 mutant strain associated with ZDV resistance and this is the first report in Japan. PMID- 10835843 TI - A comparative study of changes in fecal flora in patients preconditioned with either amphotericin B or fluconazole for allogeneic bone marrow transplantation. AB - The present study investigated fecal flora changes in 20 patients who received either the nonabsorbable antifungal agent amphotericin B, or the absorbable antifungal agent fluconazole, with tobramycin and vancomycin as preparation for undergoing allogeneic bone marrow transplantation (BMT). The oral antibiotic regimen consisted of tobramycin (540 mg/day, three times a day), vancomycin (1,500 mg/day, three times a day) and either amphotericin B (2,400 mg/day, twice a day) (AMPH group) or fluconazole (400 mg/day, twice a day) (FLCZ group) and was designed to prevent infections from microorganisms in the gut. Aerobic bacterial colonies were not detected on the day of BMT or 1 week after BMT, except in 1 case unable to take the full antibiotic regimen due to nausea and vomiting. Anaerobic bacterial colonies were not detected on the day of BMT except in this single case. Furthermore, there were no episodes of bacterial infection. In both groups, Candida colonies were detected in some case. Candida colonies were also detected in the pharynx and urine. However, there were no fungal infections. The present report suggests that amphotericin B and fluconazole administrations, with tobramycin and vancomycin, are equally effective for protection against bacterial and fungal infections in BMT recipients. PMID- 10835844 TI - [Molecular investigation of pathogenic factors of suspected-diarrheogenic Escherichia coli isolates from patients feces]. AB - One hundred fifty-one O serotypable Escherichia coli strains which were assumed diarrheogenic E. coli among 2,240 strains of E. coli isolated from the in- and outpatients stools with or without gastrointestinal symptoms at Kyorin University Hospital from February 1994 to September 1996 were examined for the relationship between the possession of eight pathogenic factor-related genes and gastrointestinal symptoms of the patients using the polymerase chain reaction (PCR) for these strains. The rate of possession of pathogenic factor-related genes in the E. coli examined was 20.5% (31 strains) and gastrointestinal symptoms were found in all the patients with these strains except one. In the patients without gastrointestinal symptoms, E. coli isolates that possesses these genes was detected in only one case during 61 cases. The respective genes detected were eaeA and astA in each 14 strains, VT1 in 6, VT2 in 5, ST1b in 4, aggR in 3 and LT in 2, ST1a and invE gene was not detected. In particular, the O157 strains were found in 55.6% (5/9 strains) for these genes, and individual strains had VT1, VT2, eaeA and astA genes simultaneously. In contrast, none of these related genes was found in 9 strains of enteroinvasive serotype but enteropathogenic E. coli-related genes were found in 3 strains. The rate of possession of the genes related to enterotoxigenic E. coli, O159 which was most frequently isolated was low as 2.3% (1/43 strains, astA gene) and there were strains showing low correlation to the state of possession of the genes with the O serotype. Since the prevalence of the gastrointestinal symptoms is clearly high for the case which possesses the strain of which the pathogenic factor-related gene was detected, it was suggested that detection of pathogenic factor-related genes in E. coli isolates from feces using the PCR could be an effective means to decide whether the bacteria concerned was a causal bacteria or not in clinical practice. PMID- 10835845 TI - [Deep-seated mycosis in patients with solid-organ transplantation--a study of autopsied cases in the United States]. AB - Autopsies with solid-organ trans plantation (SOT) in UCLA Medical Center were reviewed, especially focussing on the deep-seated fungal infection and the incidence of fungal infections, causative fungi, and organs involved were evaluated. Deep-stated fungal infections were demonstrated in 21.0% of the patients with SOT. The incidence of fungal infections were 26.1% in kidney transplantation, which was the highest rate in SOT autopsies we reviewed, 25.0% in liver transplantation, 14.3% in lung transplantation, and 13.2% in heart transplantation. And the most common causative fungi were Aspergillus species, seen in 70.6% of SOT autopsies. In contrast, Candida species were observed in 25.5%. In addition, the incidence of deep-seated mycosis has been increasing since 1992, this was parallel to the increase of aspergillosis, most of which were found as a generalized spreading. In contrast, candidal lesions, were almost limited to the digestive tracts. The above suggests that from this standpoint, improvement of prophylactic and therapeutic technique against aspergillosis is the greatest problem in deep-seated mycosis in SOT patients. PMID- 10835846 TI - [Clinical evaluation of commercial serological test for Bartonella infection]. AB - We evaluated the usefulness of a serological diagnostic kit (Bartonella IFA IgG, IgM; MRL Diagnostics) for Bartonella henselae infection. Of the 110 healthy individuals, 107 (97.3%) were with titers being less than 1:64 for IgG antibody to B. henselae, 2 were with titers being 1:64 and 1 with 1:128, IgM antibody to B. henselae was negative in all individuals. Serological diagnosis of cat scratch disease (CSD) using indirect fluorescence antibody (IFA) methods (in-house and diagnostic kit) was made in either elevated titers of IgM (> or = 1:20) or IgG (> or = 1:256) antibodies, or a four-fold rise in IgG titer between acute and convalescent sera. Of the 18 individuals with serological diagnosis of CSD by in house IFA method in 26 CSD clinical diagnosed patients, 15 (83%) were compatible with the results of the diagnostic kit, whereas 3 (17%) were not compatible. Of the 8 without serological diagnosis, 1 (13%) was serologically diagnosed as CSD, and the others were negative. Overall, the serological diagnosis was made in 16 of 26 (62%). The specificity and sensitivity of this kit were 100% and 62%, respectively. The cross-reaction between B. henselae and Bartonella quintana was observed in sera from controls and patients. Our results show that the diagnostic kit as well as in-house method is an useful tool for the serological diagnosis of cat scratch disease. PMID- 10835847 TI - [A neonatal varicella case confirmed early to be infected with varicella-zoster virus using PCR analysis]. AB - A neonatal varicella case confirmed early to be infected with varicella-zoster virus (VZV) using PCR analysis was reported. The mother developed an eruption on the following day the patient's birth and was diagnosed as varicella at 3 day of age. The neonate received immunoglobulin and acyclovir (17.5 mg x twice). She was speculated to develop varicella on the 5 to 10 day of age. Eruptions were noted, however, on the 13 day of age, although she was confirmed to be infected with VZV using PCR on the 6 day of age, and her symptoms were much milder than that foreseen. Delay of development and mildness of her symptoms may be attributed to immunoglobulin and acyclovir given on the 3 day of age. PCR method was very useful in determining early that she was infected with VZV. PMID- 10835848 TI - National Population Policy 2000. PMID- 10835849 TI - Autologous bone marrow transplantation. PMID- 10835850 TI - Methaemoglobinaemia in areas with high nitrate concentration in drinking water. AB - BACKGROUND: An epidemiological investigation was undertaken in all age groups to assess the prevalence of methaemoglobinaemia in areas with high nitrate concentration in drinking water. METHODS: Five areas were selected with an average nitrate concentration (as nitrate) of 26, 45, 95, 222 and 459 mg nitrate ions/litre in drinking water. These areas were visited and the house schedule (containing name, age, sex and weight of the family members) prepared in accordance with the statistically designed protocol. In all, 178 persons, matched for age and weight, were selected and arranged in five age groups. They constituted 10% of the total population of each of these areas. A detailed history of the selected population was taken, medical examination conducted and blood samples taken to ascertain the level of methaemoglobin. The collected data were subjected to statistical analysis to ascertain a relationship between nitrate concentration and methaemoglobinaemia. RESULTS: High nitrate concentrations cause severe methaemoglobinaemia (7%-27% of Hb) in all age groups, especially in the age group of less than 1 year and above 18 years. The lower levels of methaemoglobin in the age group of 1-18 years is probably due to better reserve of cytochrome b5 reductase activity and its adaptation to increasing nitrate concentration in water to compensate for methaemoglobinaemia in this age group. CONCLUSION: We conclude that high nitrate ingestion causes methaemoglobinaemia in all age groups. Cytochrome b5 reductase activity and its adaptation with increasing water nitrate ingestion plays a role in compensating for the methaemoglobinaemia. PMID- 10835851 TI - Autologous bone marrow/stem cell transplantation: initial experience at a north Indian referral centre. AB - BACKGROUND: High-dose chemotherapy and/or radiation therapy rescued by autologous bone marrow or peripheral blood stem cells is being increasingly used for the treatment of haematological and solid malignancies. While few centres in India use this modality of therapy, the worldwide experience is encouraging. We, therefore, analysed the results of our initial experience with this therapeutic modality. METHODS: Forty-two patients [multiple myeloma (17), Hodgkin's disease (4), non-Hodgkin's lymphoma (3), chronic myeloid leukaemia (2), acute myeloid leukaemia (2), acute lymphoblastic leukaemia (2), epithelial ovarian cancer (6), breast cancer (4), primitive neuroectodermal tumour and testicular germ cell tumour (1 each)] underwent high-dose chemotherapy followed by either autologous bone marrow transplant (n = 9), peripheral blood stem cell transplant (n = 30) or both (n = 3). The indications for transplant included either advanced stage at diagnosis, other adverse prognostic indicators during the course of their disease, or relapse. The data were analysed retrospectively in December 1998 using hospital records. Follow up data of all the patients were available. RESULTS: Thirty-four of the 42 patients (81%) showed stable engraftment. Eight patients (19%) died in the early post-transplant period (day 5 to day 52 post transplant). Seven patients died due to neutropenic infections and one due to acute renal failure. Of the 34 surviving patients, 20 were alive at the time of analysis and 14 had died. All but one death in this group were due to progressive primary malignancy. The median overall survival for all patients was 17 months and for the 34 engrafted patients it was 27 months. An analysis of factors affecting survival revealed that patients with chemosensitive disease had a longer overall survival (20.9 v. 6.1 months, p = 0.04) compared to those with chemoresistant disease. CONCLUSION: Autologous bone marrow or peripheral stem cell transplantation is a feasible procedure in India with an acceptable morbidity and mortality. It should be offered more frequently to properly selected patients. PMID- 10835852 TI - Mycoplasma pneumoniae and community-acquired pneumonia. AB - BACKGROUND: Community-acquired pneumonia is an important cause of mortality and hospitalization in all age groups. In temperate climates, Mycoplasma pneumoniae is a common respiratory pathogen causing pneumonia. Information on human Mycoplasma infection in India is scarce. METHODS: We aimed to determine the frequency of Mycoplasma pneumoniae infection among patients with community acquired pneumonia in a prospective cross-sectional study. The assessment included clinical and radiological evaluation followed by microbiological evaluation for the specific pathogen. Microbiological investigations included aerobic and anaerobic blood culture, anti-Mycoplasma IgM antibody detection by gelatin particle agglutination test and ELISA, culture of respiratory tract secretions for Mycoplasma pneumoniae and other organisms, and detection of specific Mycoplasma pneumoniae antigen by indirect immunofluorescence. RESULTS: Sixty-two patients (42 men and 20 women; mean age 41.7 years) with community acquired pneumonia were investigated prospectively. They included 42 immunocompetent and 20 immunocompromised patients. Six patients had definitive evidence of Mycoplasma pneumoniae infection and an additional 16 patients had a probable diagnosis. In all, 22 (35.5%) patients with pneumonia had Mycoplasma pneumoniae infection. Of these, 12 patients belonged to the immunocompromised group and 10 to the immunocompetent group. Patients with Mycoplasma pneumoniae infection also had secondary bacterial infection as evidenced by organisms isolated from blood in 50% and from respiratory tract secretions in 68%. CONCLUSION: Community-acquired pneumonia has a polymicrobial aetiology, of which the prevalence of Mycoplasma pneumoniae is 35%. The study has two implications: (i) Mycoplasma pneumoniae infection is frequently associated with secondary bacterial infection; and (ii) initial empirical antibiotic therapy for community acquired pneumonia in India must include antibiotics with activity against Mycoplasma pneumoniae. PMID- 10835853 TI - Apoptosis: future directions in cancer therapy. AB - Cancer as a multifactorial disease results in gain of immortality due to defective apoptosis. The primary mode of cell death by apoptosis induced by various modes of treatment often fail in vivo. The in vitro environment is less complex while the in vivo environment is influenced by various external regulatory signals besides the existence of multiple, parallel and independent apoptotic pathways. Further, specific preference for an apoptotic pathway in a certain cell type would significantly alter the apoptotic responses. Identification of defects in preferred pathways and choosing alternative and potentially inducible pathways would help in deciding on apoptosis-based treatment protocols. Mechanisms involved in the execution of apoptosis may also not be unique to apoptotic pathways since similar events, possibly with strict control, do occur during mitosis. Further evaluation may yield new dimensions to apoptosis and apoptosis-based therapy. PMID- 10835854 TI - Typhoid fever vaccines. PMID- 10835855 TI - Improved results of renal transplantation. PMID- 10835856 TI - Preventing transmission of blood-borne pathogens to health care workers. AB - Health care workers are exposed to blood-borne pathogens, mainly the human immunodeficiency virus, hepatitis B virus and hepatitis C virus. Infection by these viruses leads to chronic or fatal illnesses which are expensive and difficult to treat. Individuals who harbour these viruses may be asymptomatic and hence all patients should be assumed to harbour a blood-borne pathogen. All health care workers should take adequate precautions (a set of guidelines termed 'universal precautions'). Methods of preventing transmission of blood-borne pathogens include vaccination against hepatitis B virus, following universal precautions and taking adequate post-exposure prophylaxis. PMID- 10835857 TI - Hypothesis testing (Part I). PMID- 10835858 TI - Generating finances for the chemotherapy of children with cancer. AB - BACKGROUND: To develop and evaluate a system of obtaining financial assistance for the treatment of children with solid tumours by involving individual members of society. METHODS: This prospective project was carried out at the Paediatric Solid Tumours Clinic of the Department of Paediatric Surgery, All India Institute of Medical Sciences, New Delhi, from January 1994 to December 1998. Donor families were enlisted by talking to affluent people. Families who agreed to help in this effort were told that they could 'adopt' a child for the purpose of his/her treatment and that they should purchase the prescribed chemotherapeutic drugs and give them to the family of the affected child. Therapy was started once the drug was received at the hospital. This process was repeated at each subsequent visit. RESULTS: Of the 291 children with solid tumours registered at the clinic, 45 (15.5%) received financial assistance by this method. The proportion of children receiving financial assistance increased from 8.6% in 1994 to 23% in 1998. Of all those who received assistance, 20 (44.4%) have completed therapy and are surviving, 11 are still on therapy, 12 died and 2 decided to discontinue therapy because of progressive disease. CONCLUSIONS: The advantages of this system far outweigh its disadvantages. This method of generating finances for children with cancer can be recommended to all doctors treating such children. PMID- 10835859 TI - Transitional demographic and health survey: outlook in South Africa. PMID- 10835860 TI - Measles emerges. PMID- 10835861 TI - The medical profession and pharmaceutical industry. PMID- 10835862 TI - Awareness regarding use of folic acid for prevention of congenital neural tube defects. PMID- 10835863 TI - On peer review. PMID- 10835865 TI - What is plaguing blood banking: shortage of blood or ignorance of doctors? PMID- 10835864 TI - Screening for chronic impairments. PMID- 10835866 TI - Prevalence of hepatitis E virus as a causative agent of acute hepatitis. PMID- 10835867 TI - Is BCG vaccination obsolete? PMID- 10835868 TI - Laparoscopic donor nephrectomy for India. PMID- 10835870 TI - [Dialysis indications. Are we going towards a new paradigm?]. PMID- 10835869 TI - Mycobacterium in bronchoscopic aspirate and bronchoscope: an iatrogenic infection. PMID- 10835871 TI - Decision process about options in renal therapy substitution: selection versus election. AB - No renal replacement therapy stands alone. Hemodialysis, peritoneal dialysis and transplantation each have a role to play in the care of our patients. When one fails, another can replace that modality. Patients and staff should be counselled accordingly. The responsibility of healthcare workers is to try to best match the medical condition and lifestyle of the patients with the renal replacement therapy available. Furthermore the patients should have sufficient information to be able to make these decisions wisely. PMID- 10835872 TI - Comparative survival in hemodialysis and peritoneal dialysis. PMID- 10835873 TI - The diabetic patient with renal failure. PMID- 10835874 TI - Adequacy in dialysis: intermittent versus continuous therapies. AB - A vital conceptual difference between intermittent and continuous dialysis therapies is the difference in the relationship between Kt/V urea and dietary protein intake. For a given level of protein intake the intermittent therapies require a higher Kt/V urea due to the reasons mentioned above. The recently released adequacy guidelines by DOQI for intermittent and continuous therapies are based on these assumptions. The link between adequacy targets and patient survival is well documented for an intermittent therapy like HD. For a continuous therapy like CAPD however, the evidence linking improved peritoneal clearance to better survival is not as direct. However, present consensus allows one to extrapolate results based on HD. The concept of earlier and healthier initiation of dialysis is gaining hold and incremental dialysis forms an integral aspect of the whole concept. Tools like urea kinetic modeling give us valuable insight in making mathematical projections about the timing as well as dosing of dialysis. Daily home hemodialysis is still an underutilized modality despite offering best survival figures. Hopefully, with increasing availability of better and simpler machines its use will increase. Still several questions remain unanswered. Despite availability of data in hemodialysis patients suggesting that an increased dialysis prescription leads to a better survival, optimal dialysis dose is yet to be defined. Concerns regarding methodology of such studies and conclusions thereof has been raised. Other issues relating to design of the studies, variation in dialysis delivery, use of uncontrolled historical standards and lack of patient randomization etc also need to be considered when designing such trials. Hopefully an ongoing prospective randomized trial, namely the HEMO study, looking at two precisely defined and carefully maintained dialysis prescriptions will provide some insight into adequacy of dialysis dose and survival. In diabetic patients, the relationship between outcome and dialysis dose needs to be better defined. Data relating adequacy of dialysis to outcome in a pediatric population is not available. In dialysis therapy, the Risk/Dose (R/D) function does not bear a linear relationship. This together with a lack of proof equating peritoneal to renal clearance lends some uncertainty to the validity of the recommendation that there is a linear and constant decrease in RR for std (Kt/V) [equivalent standardized Kt/V calculated from average predialysis BUN for any frequency and/or combination of intermittent and continuous dialysis ref] up to 2.3 as reported in the CANUSA study. Due to the complex nature of this problem it may be prudent to undertake a multi-center trial with std (Kt/V) prospectively randomized to either 2.0 or 2.4. This would provide a reliable database to evaluate the R/D function over this critical range of normalized peritoneal urea clearance. Likewise in PD, the postulated linearity between dialysis dose and outcome needs to be studied in a prospective randomized manner. The amount of dialysis dose required for malnourished patients, diabetic and pediatric patients needs to be better defined. The role of aggressive dialysis in reversing malnutrition needs to be studied and studies need to be done to identify the most scientific use of V in malnourished patients. Justification of a healthy start/incremental dialysis based on outcome measures needs to be established and it's cost effectiveness validated by clinical trials. Again, a prospective randomized controlled trial comparing incremental dialysis with dietary protein restriction in patients with GFR < or = 10.5 ml/min/1.73 m2 with properly defined outcome measures like morbidity, mortality, decline of GFR and quality of life needs to be conducted. Comparisons of incremental hemodialysis and incremental peritoneal dialysis need to be made especially with regard to technique survival and preservation of residual renal function (RRF). (ABSTR PMID- 10835875 TI - Cardiovascular problems in ESRD patients. PMID- 10835876 TI - [Current dilemmas and future perspectives in the treatment of renal osteodystrophy]. PMID- 10835877 TI - Nutrition in chronic renal failure. PMID- 10835878 TI - [Immunosuppression after renal transplantation]. PMID- 10835879 TI - Ethanol suppresses L-arginine-induced relaxation response of rat aorta stimulated with bacterial lipopolysaccharide. AB - Using isolated rat aortic strips without endothelium, we investigated the effect of ethanol on L-arginine-induced relaxation. L-Arginine (10(-5)-10(-3) M) produced a relaxation response in vascular strips incubated for 6 hr with bacterial lipopolysaccharide (1 microgram/ml). The relaxation was either abolished by cycloheximide (10(-5) M) and actinomycin D (10(-5) M) or diminished by polymyxin B (10 micrograms/ml), dexamethasone (10(-5) M) and nitro-L-arginine (10(-4) M) under conditions in which these inhibitors were coincubated with lipopolysaccharide. Also, L-arginine-induced relaxation was significantly inhibited when ethanol (200, 400 mM) was present together with lipopolysaccharide during the 6-hr incubation. However, when ethanol was added to the organ bath after 6-hr incubation with lipopolysaccharide, it did not inhibit the relaxation. These results suggest that the relaxation response to L-arginine is mediated by an inducible type of nitric oxide synthase, which can be induced by lipopolysaccharide, and that ethanol may attenuate this nitric oxide-mediated relaxation by inhibiting expression of inducible nitric oxide synthase. PMID- 10835880 TI - The alcohol tests for drunken criminals using psychological tests. AB - In the alcohol test for drunken criminals, we introduced Bender-Gestalt test and Rorschach test for the assessment, and examined their usefulness for the evaluation of intoxication patterns according to Binder's classification. The subjects were 24 drunken criminals who were examined by Mental Hygiene Group, Tsukuba University, for psychiatric evidences. The subjects were divided into the ordinary intoxication group (OI group) and the complicated intoxication group (CI group) on the basis of the behavioral assessment, and the psychological tests were performed before and after drinking. The following results were obtained. 1) Alcohol intoxication induced decrease in R1T, W and VIII + IX + X/R and increase of BGT scores and P%, which indicates that subjects become unable to make comprehensive and delicate responses to the external stimuli. 2) When we classified subjects into increasing and decreasing type on the pattern of changes in the BGT score from before to immediately after drinking in each subject, we found the ratio of increasing type in complicated intoxication is more than in ordinary intoxication significantly. And we found significant group x drinking interaction in F+% and At% of Rorschach test. The F+% significantly decreased only in CI group. The At% in CI group tended upward, but downward in OI group. These findings indicated that complicated intoxication reduced the subject's reality testing, while not in ordinary intoxication. 3) Comparing the effects of personality and intoxication factors in complicated intoxication, intoxication factors were considered to play primary roles. 4) We found association between high BAL and reduction of ego function and imagination, which is represented as significant peak of BALx drinking interaction in the BGT scores, M and FM + m. These observations suggest that the psychological tests as part of the alcohol tests are useful for the evaluation and research of intoxication. PMID- 10835881 TI - [Expectation of future intraocular lens development]. PMID- 10835882 TI - [Analysis of cytoskeletal proteins in rat selenium cataract using two-dimensional electrophoresis]. AB - BACKGROUND AND PURPOSE: Previous studies used sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to characterize the modifications to lens proteins during opacification in the selenite model for cataract. To analyze the protein modifications in more detail, two dimensional electrophoresis (2 DE), which is more sensitive than 1 D electrophoresis, was used. RESULTS: The results of 2 DE demonstrated rapid changes in cytoskeletal proteins including spectrin and vimentin at the earliest stages of opacification. CONCLUSION: The results suggested that the mechanism of opacification might involve changes in cytoskeletal proteins during the earliest stages of cataract formation. PMID- 10835883 TI - [The level of lipoprotein(a) in human cataractous lens]. AB - PURPOSE: Molecular features of lipoprotein(a) [Lp (a)] associated with membranes of the lens were examined with respect to the pathogenesis and progression of cataract. METHODS: Lenses were homogenized and separated into urea-soluble (US) and water-soluble (WS) fractions. Then low density lipoprotein (LDL) and high density lipoprotein (HDL) fractions were separated from US & WS fractions by flotation density gradient ultracentrifugation. LDL and HDL fractions were prepared from 14 and 54 lenses, respectively, of a group of diabetic patients with senile cataract (DM group) or a group of non-diabetic patients with senile cataract (non-DM group). Lp(a) in each fraction and in human aqueous humor was immunochemically assayed using the latex agglutination method. Molecular phenotypes of Lp(a) were identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) followed by western blotting. RESULTS: Lp (a) contents in LDL fractions were significantly higher (p < 0.05) in the DM group [11.14 +/- 0.88(mean +/- standard deviation) micrograms/lens] than in the non-DM group [5.77 +/- 2.75 micrograms/lens]. Lp(a) values in HDL fractions were higher in the DM group than in the non-DM group, although the values did not differ significantly between the two groups. Lp (a) concentration in aqueous humor was slightly higher in the DM group than in the non-DM group. Lp (a) components examined by SDS-PAGE were detected only at the origin using immunoblotting. CONCLUSION: The Lp(a) content in cataractous lenses was higher in the DM group than in the non-DM group. In contrast to the molecular features of Lp(a) in blood, those of Lp(a) in cataractous lens seemed to be high molecular weight complexes. These results suggests that impairment of LDL receptors in DM is associated with disturbance in lipid metabolism which leads to accumulation of degenerated lipoproteins and altered membrane structure. PMID- 10835884 TI - [In vitro evaluation of pressure fluctuations with differing height of the infusion bottle in phacoemulsification]. AB - PURPOSE: To investigate the influence of the height of the infusion bottle and vacuum pressure on the development of an occlusion break surge in an in vitro model of the anterior chamber of the eye. METHODS: We employed a rigid chamber device to simulate the anterior chamber of the human eye and performed phacoemulsification with a synthetic nucleus. We evaluated amplitude of pressure fluctuations in the chamber. An experimental occlusion break surge was produced at vacuum pressures of 200, 300, and 400 mmHg. For each vacuum setting, the height of the infusion bottle was preset from 60 to 90 cm at intervals of 5 cm. We performed 6 measurements for each condition. RESULTS: Increase in surge deflection amplitude was proportional to the increase in vacuum pressure (p < 0.0001). At all vacuum pressures, there was no significant relation between surge deflection amplitude and bottle height. Amplitude of pressure fluctuations in the chamber did not fall below the irrigating pressure if the bottles were kept 70 cm at a vacuum pressure of 200 mmHg, and 85 cm at a vacuum pressure of 300 mmHg. At a vacuum pressure of 400 mmHg, surge deflection amplitude always exceeded irrigating pressure, even if the bottle height of 90 cm was chosen. CONCLUSION: Increases in vacuum pressure led to increase in amplitude of occlusion break surge. Elevation of the infusion bottle did not decrease the amplitude of occlusion break surge, but increased the pressure in the chamber at the beginning of the surge and prevented the pressure in the chamber from falling below the irrigating pressure. PMID- 10835885 TI - [The influence of corneal thickness and curvature on the difference between intraocular pressure measurements obtained with a non-contact tonometer and those with a Goldmann applanation tonometer]. AB - PURPOSE: The influence of corneal thickness and curvature on the difference between intraocular pressure (IOP) measurements obtained with a non-contact tonometer (NCT) and those with a Goldmann applanation tonometer (GAT) was studied. METHODS: The corneal thickness and curvature were obtained in 230 eyes of 115 subjects. The correlation between them and ratios of measurement with NCT to that with GAT ([NCT/GAT]) were examined. RESULTS: [NCT/GAT] and corneal thickness showed a significant positive correlation (r = 0.556, p < 0.01), but, the correlation between [NCT/GAT] and the radius of corneal curvature was not statistically significant (r = -0.035, p = 0.30). CONCLUSION: The thick cornea has more influence on the measurement with NCT than GAT, because IOP is measured with NCT over a wider applanation area. The corneas with steeper curvature also cause higher corneal rigidity and produce more overestimation of NCT measurement, while they have stronger capillary attraction of the precorneal tear film for the GAT tip and also produce overestimation of GAT measurement. As a result, [NCT/GAT] was believed to be not influenced by the corneal curvature. PMID- 10835886 TI - [Simulation of night myopia in pseudophakic eyes]. AB - PURPOSE: Eyes with an intraocular lens (IOL) implanted that have no accommodative ability are likely to have night myopia caused by Purkinje's shift and chromatic aberration. We evaluated the changes in retinal images caused by night myopia in various IOL-implanted eyes by simulation using model eyes. METHODS: A polymethylmethacrylate (PMMA) IOL, soft acrylic IOLs, and high-refractive-index silicone IOLs were prepared, and inserted into the model eye. The image plane of the model eye was determined as the line image at which a slit light of 560 nm was best focused, which was regarded as emmetropic condition under photopic vision. A slit light of 505 nm was then directed into the model eye. Its line image on the image plane was blurred, which was regarded as myopic condition under scotopic vision. Under each condition, the modulation transfer functions (MTF) were calculated and the line images were photographed. RESULTS: The degree of reduction of MTF and blurring of the line image under the night myopic condition was in the order of PMMA < soft acrylic < high refractive index silicone IOL. CONCLUSION: Eyes with a soft acrylic or high-refractive-index silicone IOL may have more intense night myopia caused by chromatic aberration. PMID- 10835887 TI - [Molecular biological analysis of retinal and streptococcal heat-shock protein 60]. AB - PURPOSE: The observation of recurrent oral aphtha preceding ocular inflammation in patients with Behcet's disease suggests a role of oral Streptococcus in the etiology of this disease. Heat-treated Streptococcus antigen can induce ocular inflammation or systemic symptoms in Behcet's disease patients. Furthermore, the presence of an autoantibody against retinal heat-shock protein 60 (HSP 60) has been detected in the sera of these patients. Injection of extracted retinal HSP 60 also induces experimental uveitis in treated rats. The characteristics of retinal HSP 60 and HSP 60 from S. pyogenes were evaluated using a molecular biological approach. METHODS: The gene encoding HSP 60 was isolated by polymerase chain reaction (PCR) from a bovine retinal cDNA library and from S. pyogenes DNA. The DNA sequence of the HSP 60 coding region was determined, and the amino acid sequences of the encoded proteins were predicted from the DNA sequence. The synthetic peptide (50 nmol) was emulsified with Freund's complete adjuvant and injected into rats. RESULTS: Comparison of the amino acid sequences of Streptococcal and bovine retinal HSP 60 revealed about 200 residue regions with 47% homology. Experimental uveitis was mainly induced in rats inoculated with retinal HSP 60-derived peptide or Streptococcus HSP 60-derived peptide that is equivalent to residues 245-259 of human HSP 65. DISCUSSION: The retinal and Streptococcal HSP 60 amino acid composition is thought to be important and useful for investigating the mechanisms involved in the induction of Behcet's disease. PMID- 10835888 TI - [Effects of different types of disparity cues on the response of axis-orientation selective cells in the monkey parietal cortex]. AB - PURPOSE: To specify the cues for the discrimination of orientation in depth in axis orientation selective (AOS) neurons. METHODS: We analyzed the responses of AOS neurons in the monkey caudal intraparietal sulcus (cIPS) region using binocular disparity stimuli generated by stereoscopic 3 D computer graphics. RESULTS: Most AOS neurons (20/27) were sensitive to binocular disparity and showed tuning to the orientation of a slit in the sagittal plane with orientation disparity cues. For 12 neurons we also used an array of discs or dots instead of slits to eliminate orientation disparity. Half of the neurons (6/12) responded better to the slits than to the discs or dots, suggesting that they were sensitive to orientation disparity. Five neurons (5/12) responded equally well to the discs or dots suggesting that they were more sensitive to the gradient of horizontal disparity than to the orientation disparity. CONCLUSION: Both orientation disparity and disparity gradient were likely to be integrated in the cIPS area to represent axis orientation of an object in space. PMID- 10835889 TI - [Ciliochoroidal detachment following scleral buckling surgery for rhegmatogenous retinal detachment]. AB - PURPOSE AND METHODS: We observed the peripheral choroid; ciliary body, and depth of the anterior chamber by ultrasound biomicroscopy (UBM) in 31 eyes with rhegmatogenous retinal detachment before and after scleral buckling surgery. Scleral encircling was performed in 11 eyes and segmental scleral buckling in 20 eyes. RESULTS: With UBM, ciliochoroidal detachment was detected in all eyes (100%) following scleral encircling and in 8 eyes (40.0%) following segmental scleral buckling. After scleral encircling procedure, the eyes with preoperatively bullous and wide retinal detachment showed a severe ciliochoroidal detachment and edema of the ciliary body. Shallowing of the anterior camber occurred in all 11 eyes (100%) after scleral encircling and in 12 of 20 eyes (60.0%) after segmental scleral buckling. Marked shallowing with closure of the angle and elevated intraocular pressure occurred in 2 eyes. CONCLUSION: The results showed that careful postoperative examinations for the anterior segments, chamber angle, and intraocular pressure are necessary with slit-lamp examination and applanation tonometry after scleral buckling surgery. PMID- 10835890 TI - [Clinical and experimental observation of glistening in acrylic intraocular lenses]. AB - PURPOSE: To study whether the glistening on implanted acrylic intracular lenses increases or not. METHOD: 1. Prospective study: We studied the glistening on 49 implanted acrylic intraocular lenses, particularly the time of appearance and increase after glistening appeared. 2. EXPERIMENT: We kept an intraocular lens immersed in saline solution at 50 degrees centigrade for two hours and then moved this lens into saline solution at 35 degrees centigrade, and observed this lens during immersion. RESULTS: 1. Prospective study: Glistening was present in 28 eyes (57%), and appeared from 2 to 16 months postoperatively. A few months after appearance it become stable and did not increase further. 2. EXPERIMENT: On the 10th day after observation, we could see glistening on the lenses. The points of glistening were 10 microns or less in size, and did not change during 60 days of observation. CONCLUSION: The glistening in acrylic intraocular lenses seemed to become stable a few months after appearance. This our experimental method is useful, because it gives us glistening similar to clinical cases in a short time. PMID- 10835891 TI - [Expression of the varicella zoster virus thymidine kinase and cytokines in patients with acute retinal necrosis syndrome]. AB - PURPOSE: Acute retinal necrosis (ARN) is caused by varicella zoster virus (VZV) infection. In this study, we investigated the activity of this virus and expressions of some cytokines. PATIENTS AND METHODS: The expression of VZV thymidine kinase and some cytokines were investigated by reverse transcriptase polymerase chain reaction (RT-PCR) in 9 eyes of 8 patients with ARN. RESULTS: Thymidine kinase expression was observed in all samples except one. Several cytokines, such as interferon (IFN) gamma, tumor necrosis factor (TNF) alpha, interleukin (IL)-1 beta, IL-6, and transforming growth factor (TGF) beta 1 were observed in the samples. Among these cytokines, a statistically significant expression of IFN gamma was observed in the samples of ARN, when compared to those of proliferative vitreoretinopathy (PVR) or other uveitis. The expression of IFN gamma also decreased during successive follow-ups. CONCLUSION: These cytokines may play an important role in the immune response in ARN. PMID- 10835892 TI - [Subjective burdens of families caring for "frail elderly" in their homes]. AB - The purpose of the study is to measure the subjective burdens of families caring for "frail elderly" in their homes and to clarify the relationships between caregivers' subjective burdens and the health conditions of the "frail elderly", the family's conditions, the use of social services and availability of social supports. The subjects consisted of caregivers of the elderly who also were using nursing services from K Home Visit Nursing Care Station (K-Station). We mailed subjective burden questionnaires to 98 families among users of K-Station from December of 1994 to March of 1995. We used the 12 items developed by Nakatani as a scale to measure subjective burdens. Each item was stratified into 4 grade levels from "agree" to "disagree". We asked nurses at K-Station about "frail elderly" ADL conditions in detail, the extent of medical dependency and the will of families to care for the elderly. A total of 76 questionnaires were returned, of which 73 could be analyzed. The greater part of "frail elderly" were female (n = 42). The average age of the "frail elderly" was 81.1(+/- 7.6) years old. All caregivers were members of the family of the "frail elderly" and most of them were women (n = 63). From the result of principal analysis, 2 principles were extracted from the 12 items used to measure subjective burden of caregivers. We analyzed the first principle as "subjective burden of caregivers". From the result of multiple regression analysis (Stepwise method), "subjective burden of caregivers" was characterized by the caregivers' conditions only (caregivers' health status, the will to care for the elderly and anxiety of providing night care). This result suggested that support for the caregivers of the "frail elderly" was important to sustain in-home care. PMID- 10835893 TI - [Epidemiological study on idiopathic scoliosis in high school students. Prevalence and relation to physique, physical strength and motor ability]. AB - OBJECTIVE: We applied the tuberculosis examination radiographs to scoliosis screening in high schools. Prevalence, physique, physical strength and motor ability of the students with scoliosis were investigated. METHODS: In this study, 3,299 the first year high school students (1,597 males and 1,702 females) in Wakayama Prefecture, who had taken chest X-ray photographs between 1993 and 1995, were examined and 106 cases (22 males and 84 females) were identified with scoliosis of more than 10 degrees Cobb angle. Each of the cases was assigned age- and sex-matched controls without scoliosis. These pairs were investigated as to the following items: body height, body weight, sitting height, chest girth, leg length, body mass index, vertical jump, side-to-side jump, back strength, grasping power, trunk extension, standing trunk flexibility, 50 m dash, long jump, handball throwing, and chinning. RESULTS: The body weight, chest girth, and body mass index was significantly lower in girls with scoliosis. As to physical strength and motor ability, only standing trunk flexibility was significantly lower in girls with scoliosis. CONCLUSION: Tuberculosis examination radiographs were useful for scoliosis screening in high schools. We found that girls with scoliosis were thinner. We think it is necessary for school teachers to pay more attention to their students' life style. PMID- 10835894 TI - [Blood pressure measurement by primary care physicians: comparison with the standard method]. AB - OBJECT: To examine the usual methods of blood pressure (BP) measurement by primary care physicians and to compare them with the standard methods. METHOD: DESIGN: Cross-sectional survey by self-administered questionnaire. SUBJECTS: Primary care physicians who graduated from Jichi Medical School and were working at clinics. Each standard method for 20 items was defined as the one that was most frequently recommended by 6 guidelines (USA 3, UK 1, Canada 1, Japan 1) and a recent comprehensive review about BP measurement. RESULTS: Of 333 physicians, 190 (58%) responded (median age 33, range 26 to 45 years). Standard methods and percentages of physicians who follow them are: [BP measurement, 17 items] supported arm 96%; measurement to 2 mmHg 91%; sitting position 86%; mercury sphygmomanometer 83%; waiting > or = 1 minute between readings 58%; palpation to assess systolic BP before auscultation 57%; check accuracy of home BP monitor 56%; Korotkoff Phase V for diastolic BP 51%; bilateral measurements on initial visit 44%; small cuff available 41%; > or = 2 readings in patients with atrial fibrillation 38%; > or = 2 readings on one visit 20%; cuff deflation rate of 2 mmHg/pulse 14%; large cuff available 13%; check accuracy of monitor used for home visit 8%; waiting time > or = 5 minute 3%; readings from the arm with the higher BP 1%. [Knowledge about BP monitor, 2 items] appropriate size bladder: length 11%; width 11%. [Check of sphygmomanometer for leakage, inflate to 200 mmHg then close valve for 1 minute] leakage < 2 mmHg 6%; median 10 (range 0-200) mmHg. Average percentage of all 20 items was 39%. Number of methods physicians follow as standard: median 8 (range 4 to 15) and this number did not correlate with any background characteristics of the physicians. Furthermore, we also obtained information on methods not compared with the standard. Fifty-four percentage of physicians used more standard methods in deciding the start or change of treatment than in measuring BP of patients with good control. About 80% of physicians use home BP readings in diagnosis or treatment of hypertension, but about half of physicians with ambulatory BP monitors use their measured readings. CONCLUSION: Primary care physicians used various techniques for routine BP measurement and no physician completely followed the standard. Such measurements may affect the diagnosis and treatment of hypertension, but measuring all BPs solely by the standard is not practical. We need to have a practical and efficient method of BP measurement for routine practice in the primary care setting. PMID- 10835895 TI - [Characteristics of dropouts and participants in a twelve-year longitudinal research of Japanese elderly]. AB - PURPOSES: The attrition of respondents in panel studies of the elderly can create bias in data analysis. The purposes of this study are two fold; 1) to examine characteristics of dropouts, from a particular panel lost except due to natural attrition (by death) by comparison with continuing participants in that wave, and 2) to assess representativeness of those actually continuing in a particular panel by comparison with those eligible for inclusion in that wave. Only those who died were excluded from the group of respondents at the baseline survey because they constituted natural attrition in this longitudinal survey. METHOD: At baseline (1987), 2,200 individuals age 60+ from 3,288 national representative sample were interviewed. Non-response status to three contacts (1990, 1993, 1996) with the panel was examined in relation to variables included in the baseline interview. A number of characteristics of demographic background, health, life style, and social relations obtained in the baseline survey (1987) were compared between those re-interviewed in a particular panel and subjects lost through unnatural attrition until that wave. To study the influence of unnatural attrition on variable distributions and each related factors of two health indicators (self-rated health and depressive symptoms), baseline responses were compared between those re-interviewed in a particular panel and all who were eligible to respond in that wave. RESULTS: 1) Dropouts lost to each wave were significantly older and had a lower level of social participation than persons remaining in that wave. Significant differences in health and life-style variables appeared between dropouts lost and continuing participants until third or later waves. 2) Continuing participants in a particular panel were likely to be younger, to be more physically, mentally, or socially healthy than those eligible to respond in the wave. Each related factors of two health indicators were almost same between those re-interviewed in a particular panel and those eligible to respond in that wave. CONCLUSION: Dropouts in longitudinal research were found to appear nonrandomly. While distributions of age and health indicators in those re-interviewed were influenced by respondent attrition, related factors of health indicators may be free of bias that can be created by it. PMID- 10835896 TI - [Outcome based quality-improvement methods in visiting nursing services (VNS) in Japan: practical report and feasibility]. AB - For the execution of the nation's public long-term care insurance in the year 2000, and Quality Improvement (QI) of visiting nursing services (VNS) will become extremely important in Japan. Some standard methods to evaluate structure or process in VNS have been published but there is no report of actual QI performance based on outcome measurement in each VNS institution. Modifying the method introduced by JCHO in USA, we performed QI using one negative outcome "unexpected cessation of home care" as a monitoring outcome measurement. In three VNS institutions variously located in Japan, cases of unexpected-cessation in one year were monitored retrospectively, then blindly assessed by nurses. Cases were categorized into "avoidable cessation" or "unavoidable unexpected-cessation", and for the "avoidable" cases the kinds of improvement that would be necessary to avoid this type of unexpected-cessation in the future were discussed. Depending on the situation of the facility, some of the suggested improvements were executed within possible ranges. The numbers of unexpected-cessation of home health care in each institution were 89, 58 and 15 with incidences from all the cases in one year of 46%, 40% and 42% respectively. In these cases, 47, 39 and 15 cases were categorized as "avoidable". Generally, improper nursing assessment in changes of patients' physical condition was the biggest cause of home health care unexpected-cessation. Although some institution were not able to accomplish any part, each institution were able to capture their unique problems and made a detailed specific QI plan to avoid cessation of home care in the future. The two main problems in feasibility were "the analysis took too long (8-9 hours)" and "part-time staff member could not participate". This simple modified QI method provides useful Quality Improvement of Japanese VNS with a limited staff. However, to facilitate execution, a much fairer recognition of the importance of QI, computerization or the partial consignment to a third party, and investigation over a shorter period were considered to be effective. PMID- 10835897 TI - [Does disclosure of medical records to patients influence medical care?]. AB - Disclosure of medical records to patients has been assessed, but the influence of disclosure on medical care has not been well researched. To address this situation, this study was conducted to test three hypotheses: 1) doctors think that the disclosure can influence medical care, 2) whether doctors think disclosure can influence medical care depends on how they rank medical records (for example, as evidence for diagnosis and treatment, a process to reach a diagnosis, a tool to communicate with other medical staff, etc), and on whether they think medical record disclosure could change the content of the record. Questionnaires were sent to 881 doctors who work at two hospitals affiliated with N Medical School and K Medical University. RESULTS: Four hundred eighty-eight doctors responded. The findings related to the hypotheses were as follows: 1) Those who answered that the disclosure could have an influence on medical care were 49.1% of the total. 2) There was no relation between how doctors ranked medical records and whether they thought disclosure could influence medical care. 3) Doctors who answered that there were things that they would not be able to write if medical records were disclosed accounted for 73.5% of the total. These doctors answered "yes" significantly higher to the question that the disclosure could influence medical care than others (Odds 3.6, P < 0.01). Doctors who thought they would not be able to enter the diagnosis, the name of the disease for insurance, self-evaluation, information that could be judged and subjective information answered that disclosure could influence on medical care (P < 0.05). It was assumed that disclosure of medical records to patients could change the content of the medical records and that could influence medical care. How to deal with information will become an important issue if records are disclosed. PMID- 10835898 TI - Energy regulation and aging: recent findings and their implications. AB - Old age is a time of vulnerability to unintentional weight loss, a factor that is associated with increased morbidity and premature death. Many possible causes of weight loss in old age have been suggested. The so-called anorexia of aging may play a particular role, by either reducing food intake directly or reducing food intake in response to such adverse factors as age-associated reductions in taste and smell, poor dentition, use of multiple prescription medicines, and depression. Recent studies also raise the question of whether a reduction in dietary variety may be important. These findings emphasize the need for regular monitoring of body weight to detect unintentional weight loss in older individuals and suggest testable ways to minimize the impact of the anorexia of aging on body weight through improved dietary management. PMID- 10835899 TI - Dieting, essential fatty acid intake, and depression. AB - Studies have both supported and contested the proposition that lowering plasma cholesterol by diet and medications contributes to depression. Evidence suggests that an imbalance in the ratio of the essential fatty acids (EFAs), namely the n 6 and n-3 fatty acids, and/or a deficiency in n-3 fatty acids, may be responsible for the heightened depressive symptoms associated with low plasma cholesterol. These relationships may explain the inconsistent findings in trials of cholesterol-lowering interventions and depression. Dieting behaviors have similarly been associated with alterations in mood state. This review discusses studies of EFAs and depression and explores the involvement of dieting-related psychological factors as potential confounders. PMID- 10835900 TI - Vitamin E and memory: is it vascular protection? AB - Vitamin E per unit serum cholesterol has been found to correlate with memory performance in a recent analysis of data from the Third National Health and Nutrition Examination Survey. Because vitamin E is thought to be protective against vascular disease, this finding is consistent with the growing body of evidence that implicates vascular disease and vascular disease risk factors in age-related cognitive decline and Alzheimer's disease. PMID- 10835901 TI - Should dietary calcium and protein be restricted in patients with nephrolithiasis? AB - Renal stone disease is a painful condition that affects 1-20% of the general population. Therapy aimed at decreasing the incidence of recurrent stones includes dietary advice. Dietary considerations include intake of both calcium and protein. Calcium restriction in stone formers is not recommended because it can have adverse effects on bone and the incidence of stones. Although a high protein diet can elevate urinary calcium, uric acid, and sulfate and decrease urinary citrate, which may alter the propensity to form stones, restriction of protein to less than the current RDA for the management of stone disease can not be recommended at this time. PMID- 10835902 TI - Noah grows up: transitioning problems from special feeding routes to oral intake. AB - As more children with special health care needs survive into adulthood, there is an increased need for help with the transition from special feeding routes to oral intakes. In infancy and early childhood, emphasis is often placed on the medical aspects of treatment for these children. As they grow and integrate more fully into the community, attention shifts to their educational needs. Throughout life, however, the need for attention to nutrition issues and needs remains. A case study illustrates the experiences of a young man with special oral intake needs; feeding and related transitions were required for independent functioning as he matured into a young adult. His many struggles are summarized, nutrition implications are outlined, and recommendations are provided for dealing with similar patients. PMID- 10835903 TI - [Silent myocardial ischemia]. AB - Silent myocardial ischemia (SMI) is a manifestation of coronary artery disease in which persons have episodes of myocardial ischemia that are not accompanied by chest or anginal pain. Episodes of SMI have been described as the predominant form of transient myocardial ischemia in patients with stable angina, those with unstable rest angina and even variant angina. Silent myocardial ischemia is also known to occur in some patients with myocardial infarctions and resuscitated from episodes of sudden cardiac death, thus occur frequently in patients with all of the usual symptomatic phases of coronary artery disease. A number of studies have now clearly demonstrated that patients with silent ischemia have increased risk for coronary events: sudden death or myocardial infarction. SMI is most often recognized during exercise testing or fortuitously with ambulatory electrocardiography. For today it is imperative to investigate every patient in whom multiple risk factors of IHD have been recognised, even in the absence of chest pain. The early detection of SMI may help to prevent later cardiac events. If the ischemia (symptomatic and silent) itself is the cause of the higher mortality rate, then the goal of therapy should be to abolish it either by pharmacological means or by invasive interventions as balloon angioplasty or coronary bypass surgery. PMID- 10835904 TI - [Effect of coffee on blood pressure and activity renin-angiotensin- aldosterone system and catecholamines concentration in patients with essential hypertension]. AB - The purpose of this study was to investigate whether the renin-angiotensin aldosterone system activity RAAA or plasma catecholamines concentrations and blood pressure are related to distinct patterns of drinking one cup of coffee in hypertensive patients with low or normal plasma renin activity. Plasma renin activity and enzyme converting angiotensin I to angiotensin II activity and serum concentration of aldosterone and catecholamines were measured by radioimmunoassay. Blood pressure was measured by ambulatory monitoring. Our data suggest that drinking one cup of coffee caused after 60 min, to 2 hours elevated systolic and after 60 min diastolic pressure, only in hypertensive patients, with low RAAA habitually drinking coffee. Hypertensive patients with normal RAAA, habitually drinking coffee, after drinking one cup of coffee had elevation only of diastolic blood pressure from 60 min to 120 min after drinking, but systolic blood pressure did not elevate. Plasma catecholamines and aldosterone concentrations, blood renin activity and enzyme converting angiotensin I to angiotensin II activity were not elevated in hypertensive patients with low or normal RAAA after drinking one cup of coffee. PMID- 10835905 TI - [Side effects during dobutamine stress echocardiography: analysis of 582 studies]. AB - The purpose of the study was to assess the safety, adverse effects and complications of the dobutamine stress echocardiography (ED). 582 patients without previous infarction were prospectively studied with ED. There were 196 female and 368 male, age varied from 27 to 74 years, mean 52. Dobutamine was given in stepwise increasing doses from 5 to 40 mcg/kg/min. Mean maximal dose achieved was 33 mcg/kg/min. Atropine was added in 253 (43%) cases. Significant coronary artery disease was present in 323 patients (53%). There were no death, no myocardial infarction or episodes of sustained ventricular tachycardia as a result of ED. The test was terminated when following conditions were revealed: target heart rate (28.9%), maximal established dose achieved (25.3%), left ventricular asynergy (19.6%), angina pectoris (10.8%), increase of systolic blood pressure above 220 mm Hg (2.6%), hypotension (7.6%), nonsustained ventricular tachycardia (1.7%). The most common non-cardiac side effects were skin tingling (19.8%), atypical chest pain(16.3%), palpitations (13.9%) and headache (7.9%). The most side effects were usually well tolerated, without the need for test cessation. The ED was terminated only in 4 (0.6%) patients because of non-cardiac side effects including nausea (0.3%) and headache (0.3%). We conclude that ED may be safely performed in routine clinical practice. Side effects were rare and usually minor. Most severe ischemic pain was relieved by test interruption and sublingual nitro-glycerine or short acting beta-blocker administration. PMID- 10835906 TI - [Simple vs bifurcated graft operations in patients with abdominal aortic aneurysm]. AB - We analysed 148 patients treated surgically from 1992-1997 because of abdominal aortic aneurysm (AAA). All patients were divided into two groups: group I--118 (79.7%) patients operated with the implantation of the straight graft and group II--30(20.3%) with the bifurcated graft. The aim of the study was to analyse early (30 days after operation) complications in both groups. The mortality rate was 8.5% for group I and 26.7% for group II. The mortality was also analysed according to AAA symptoms (asymptomatic, symptomatic and ruptured). The death rate in asymptomatic patients from group I was 1.1% versus 13.3% in group II. In symptomatic patients the difference was not statistically significant--20% in group I versus 22.2% in group II. The mortality rate in patients with ruptured AAA was 50% for group I and 66.7% for group II. The early morbidity rate was significantly higher in the second group(p < 0.05) although coexisting diseases were similar for both groups. According to our material we conclude, that operation of AAA should be finished in abdomen if there is only technical possibility. It is especially important for patients with ruptured AAA. PMID- 10835907 TI - [Complication risk after abdominal aortic aneurysm operations]. AB - Between 1992-1997 185 patients were treated in our Department because of abdominal aortic aneurysm (AAA). The aim of the study was the evaluation of frequency of hospital mortality (30 days) in patients treated because of AAA. One hundred forty eight (80%) patients were operated on and 37 (20%) were treated classically. The surgical group of 148 patients were divided into three parts: group I: 106 patients with asymptomatic AAA, group II: 24 patients with symptomatic AAA and group III: 18 patients with ruptured AAA. Straight graft was performed in 118 patients (79.7%) with hospital mortality rate 8.5% and bifurcated graft implanted in 30 patients with mortality rate 26.7%. Analysis of our material allowed to find that hospital mortality was in group I: 2.8%, group II: 20.8% and in group III: 55.6%. The main cause of deaths in patients from groups I and II was myocardial infarction and hypovolemic shock in group III. The most common postoperative general complications were cardiac and pulmonary and were significantly more often in group III (p < .05). Other not significantly registered postoperative complications as renal insufficiency were also more common in patients from group III. Analysis of our material revealed that patients with abdominal aortic aneurysm should be operated selectively, when aneurysm diameter reaches 50 mm, and optimal method is straight graft which allows to reduce to minimum postoperative complications. PMID- 10835908 TI - [Endoscopic ultrasonography in the diagnosis of unexplained extrahepatic cholestasis]. AB - The aim of this prospective study was to assess the diagnostic value of EUS for extrahepatic cholestasis unexplained before by US, ERCP and CT. Between 1994-1997 we examined 50 patients (24 women and 26 men, age range 21 to 72 years, mean age 53.2 years) with clinical and biochemical parameters of cholestasis, using the ultrasound endoscope with a 7.5 MHz linear electronic array transducer (Pentax FG 32UA/Hitachi 405EUB console). In one patient EUS was normal. In other 19 patients the causes of cholestasis were neoplasms, in 22 patients--inflammatory diseases and in 8 patients--choledocholithiasis. In majority of patients with chronic calcifying pancreatitis anicteric cholestasis was observed. 20 patients were operated on. Endo-sonographic diagnosis was confirmed in all patients with neoplasm of Vater papilla and pancreas, chronic calcifying pancreatitis and pancreatic abscess. Choledocholithiasis were confirmed by endoscopic sphincterotomy in all but one patient, endoscopic biliary drainages were performed in 9 patients. In 4 cases the diagnosis was false, mainly in patients with pseudotumorous pancreatitis. No complications during EUS were observed. EUS is efficacious and safe method in the diagnostics of extrahepatic cholestasis. In some cases it can replace diagnostic ERCP. PMID- 10835909 TI - [Evaluation of cardiovascular and sudomotor functions in Alzheimer's disease]. AB - The assessment of some aspects of cardiovascular and sudomotor system was performed in the group of patients with Alzheimer's disease. No one of patients complained of disturbances arising from these two parts of the vegetative system. The autonomic functions were evaluated by means of: clinical examination, heart rate variability tests at rest, measures of blood pressure and heart rate in supine and standing position and skin sympathetic response. At rest heart rate and its variability as well as blood pressure in the patients did not differ significantly from the results obtained in the healthy controls. On standing, changes in systolic blood pressure and heart rate were recorded in the controls, but not found in the patients. The majority of patients presented with abnormalities in skin sympathetic response: lack of response or prolongation of its latency. The results of motor and sensory nerve conduction studies excluded neuropathic changes in the patients. We conclude that autonomic disturbances accompanied Alzheimer's disease and were probably caused by damaged control mechanisms within the central vegetative system. PMID- 10835910 TI - [Surgical management in the non-neoplastic and neoplastic thyroid goiter with Hurthla cells]. AB - The routine preoperative cytological evaluation of pathological changes of the thyroid more and more frequently reveals single or multiple Hurthl cells in the material obtained by the method of fine-needle aspiration biopsy (FNA). Hurthl cells are a degenerative form of epithelial follicular cells of the thyroid and cytological examination is insufficient for unequivocal differentiation between adenoma and carcinoma. Management of patients with FNA-diagnosed Hurthl cells arouses a number of controversies. Therefore, the authors of the present study subjected the thyroids of patients with cytologically diagnosed Hurthl cells to postoperative histopathological analysis. Appropriate surgical management was indicated. PMID- 10835911 TI - [Laparoscopic adrenalectomy from the lateral retroperitoneal approach]. AB - From April to November 1998 eleven patients (8 female and 3 male, mean age 48.5; range 23-69) with adrenal gland tumor underwent laparoscopic adrenalectomy from the lateral retroperitoneal approach. Authors report their primary experience; introduce their own material and describe the operative technique of laparoscopic adrenalectomy from lateral retroperitoneal approach. Besides one case of Conn syndrome, all tumors were hormonally non-active. In 10 patients the adrenal masses were incidentally discovered during such imaging examination as abdominal ultrasonography and computed tomography. All tumors were benign in postoperative histopathological examination. The localization was unilateral and suprarenal in all cases (7 right, 4 left). Both operation and postoperative course in all patients were uncomplicated. One conversion to "open" procedure had been performed due to laparoscopic equipment damage. PMID- 10835912 TI - [Common double inlet ventricle (univentricular heart) in 27 year old man: a case report]. AB - We present 27-years old man with common double inlet ventricle (CV), hypoplastic pulmonary artery and transposition of the great arteries which was diagnosed in childhood. Based on echocardiographic examination we found unidentify type of CV. Patient did not have surgery and was in good condition. He has been working. We believe that main reason of his good condition is hemodynamically optimal width of the hypoplastic pulmonary artery. PMID- 10835913 TI - [Acute intermittent porphyria and simulated acute porphyria: case reports]. AB - The authors present according to a case of acute intermittent porphyria, diagnostic and therapeutical problems, which are met by physicians in a regional hospital (case 1). Simultaneously the authors present a case of woman with opiate depends syndrome, who has simulated attacks of acute porphyria for the last few years in order to obtain narcotic drugs (case 2). PMID- 10835914 TI - [Vasocardiogenic syncope: pathogenesis, diagnosis and treatment]. AB - Syncope is important clinical problem which is a cause of 1-2% cases of hospitalization. Annual mortality in patients with vaso-cardiogenic syncope is 30%. Vaso-cardiogenic syncope can be related to reduced cardiac output, reduced venous return or disturbations of blood flow in vessels supplying central nervous system. The basis of diagnosing of syncope is precise physical examination and anamnesis. In further diagnosis one can use: echocardiography, ecg Holter monitoring, exercise test, transesophageal atrial stimulation, tilt-up test, electrophysiologic examination. Treatment of syncope consists in elimination of causes or factors inducing syncope. PMID- 10835915 TI - [Electrically enhanced damaged tissues healing. Part II: direct and pulse current in soft tissue healing]. AB - Methodology of soft tissues wounds, ulcers and pressure sores healing with direct current is described by the authors. Results of clinical trials and animal experiments are represented, as well as technical and using data. Electrical properties of damaged tissues (e. i. skin battery, vascular-interstitial closed circuits etc.) and probable electrical healing mechanisms are discussed. Effects of electrical current on batteries are described. Inductive and capacitive coupling of electric and magnetic fields, and high voltage electrostimulation for enhance tissue healing are also described in the article. PMID- 10835916 TI - [Aging, physical activity and free radicals]. AB - A growing amount of evidence indicates that increasing with age imbalance between pro- and antioxidants is implicated in a premature ageing and a variety of diseases. Physical activity influences both the production of reactive oxygen species and physiological antioxidant defences. This paper reviews the current literature on the influence of physical exercise on the oxidative stress markers and antioxidant defence system in the elderly people. PMID- 10835917 TI - [Chronic renal failure: if and to what extent early nephrological care can diminish progression of disease?]. PMID- 10835918 TI - [Bone turnover markers in diabetics treated with continuous ambulatory peritoneal dialysis]. AB - Low turnover renal osteodystrophy (ROD) is an important problem in patients on CAPD. The aim of the study was to evaluate in CAPD patients the frequency of low turnover ROD with special interest to diabetic nephropathy (DN) as well as estimation the clinical usefulness of some biochemical bone turnover markers in monitoring this complication. The study was performed in group of 54 patients: 28 on CAPD (15 M, 13 F) and 26 on HD (14 M, 12 F). There were 20/28 patients with DN in CAPD group. The observation lasted 12 months. Every 3rd month levels of iPTH, PICP, PINP, ICTP, Ca, P, total AP in serum were determined. Correlation between these biochemical markers of ROD and causes of renal failure, sex, age of patients were analyzed statistically. In CAPD patients significantly lower values of iPTH were found in comparison with HD patients (p < 0.01). In patients with DN levels of iPTH were significantly lower (p < 0.001) than in patients with other causes of renal failure (respectively: iPTH 142 +/- 81 vs 403 +/- 128 pg/ml). In patients with DN treated with CAPD levels of iPTH were significantly lower in men than in women and in younger men with DN (< 50 years). The similar results were found in levels of PINP as a marker of bone formation. Close correlation between PINP and iPTH (r = 0.80) was also found. In summary we showed that low turnover ROD is the predominant bone lesion in CAPD patients. Men with DN < 50 age are specially predisposed to the low turnover ROD. Monitoring of iPTH and PINP levels as sensitive markers of low turnover ROD might be useful in assessment of bone turnover rate in CAPD patients. PMID- 10835919 TI - [Own modification of the visualization of adhesions in the peritoneal cavity in patients treated with continuous ambulatory peritoneal dialysis]. AB - The intraperitoneal adhesions were imagined with 3 non-invasive methods such as scintigraphy, ultrasonography, computerised tomography in 25 patients (pts) on CAPD (13 females and 12 males, aged 8-72 years). Initial imaging of peritoneal cavity was performed up to 4 months after the start of CAPD and repeated every 6 12 months of further therapy. In 9 patients the adhesions where detected at the start of CAPD. Only in 4 the progression of adhesions was diagnosed as a change of the structure from single to multiple or multilocular. In all these pts, peritonitis occurred at least once in the course of CAPD. Loss of ultrafiltration resulted in transfer from CAPD to HD. In others 5 pts the consecutive images of peritoneum were stable. In 16 remaining pts the adhesions were detected in the course of CAPD programme, in 6 of these pts after peritonitis and in other 10 without known causes. In 4 pts the progression of adhesions to multiple or multilocular was detected during next 2-3 years of CAPD programme. Two of these pts had to be transferred to HD. In 12 pts the images of adhesions were stable. Together, 6 pts (24%) had to be transferred to HD, because of the progression of intraperitoneal adhesions in the course of further CAPD. In conclusion, own modification of imaging of peritoneal adhesions let us not only to estimate the influence of this complication on the intraperitoneal dialysate distribution, but also enables non-invasive monitoring of CAPD programme. PMID- 10835920 TI - [Influence of trimetazidine on echocardiography parameters and free radical stress index in coronary artery disease and end-stage renal failure patients treated by hemodialysis: preliminary communication]. AB - The aim of the study was the assessment of the influence of trimetazidine (TMZ) on echocardiography parameters and oxidative stress markers in hemodialyzed patients with coronary artery disease. The studies were carried out in 9 patients group before and after 3 months of TMZ treatment in doses 3 x 20 mg. Additionally we measured MDA concentration in the blood, as a oxidative stress marker, before and after 2 and 3 months of TMZ treatment. At the start of the study impairment of relaxation as a sign of diastolic dysfunction was detected. This abnormality persisted up to the end of the study. After 3 months of TMZ treatment we observed elevation of ejection fraction from 59.17 +/- 5.99 to 65.33 +/- 8.7% (p = 0.05). Also occurred statistically significant decrease in MDA level after 2 (0.674 +/- 0.029 vs 0.627 +/- 0.034; p < 0.001) and 3 (0.674 +/- 0.029 vs 0.567 +/- 0.063 mumol/L; p = 0.001) months of TMZ treatment. Significant improvement of the left ventricular function and decrease of MDA levels in the blood might be due to intracellular mechanisms of TMZ activity. There were no side effects of the drug in hemodialyzed patients. PMID- 10835921 TI - [Cardiovascular complications versus adequacy and peritoneal transport characteristics in CAPD patients]. AB - The aim of the study was a comparative analysis of cardiovascular status and basic parameters of adequacy and peritoneal transport in CAPD patients. 15 persons treated with CAPD over 2 years were included in the study. Cardiovascular status at the beginning and at the end of observation was assessed using the own scoring. Patients were divided into 3 groups of cardiovascular disease (CVD) progression: group I--without CDV progression, group II--with moderate CVD progression and group III--with severe CVD progression. The biochemical factors of CVD risk at the beginning and the end of observation were also measured. The adequacy and peritoneal transport parameters (Kt/V, CrCl and D/P creatinine) were calculated. Group I was characterized by the highest average Kt/V value as well as percent of Kt/V studies > or = 2.0. These values were significantly lower in group II and III. The average CrCl value was highest in group I and lowest in group II. The relatively high, significant reverse correlation between Kt/V and CVD scoring at the end of observation was revealed. Lower but significant correlation between serum albumin and CVD scoring at the end of observation was also established. The analysis of the link between CVD progression and changes of transport characteristics in studied population revealed the highest peritoneal transport in group III from the beginning of observation. The undesirable change of peritoneal transport characteristics was stated in group II and III. The results of our study let us to conclude that in observed population only high values of adequacy parameters reduced the risk of CVD progression. Kt/V seems to be better predictor of CVD progression than CrCl. The risk of CVD in CAPD patients is probably higher due to increase of peritoneal permeability. PMID- 10835922 TI - Peritonitis in continuous ambulatory peritoneal dialysis: twenty years of center's experience. AB - Twenty years of our experiences in diagnosis and treatment of peritonitis in patients treated with continuous ambulatory peritoneal dialysis are presented. Special attention was put on reduction of frequency and efficacy of treatment improvement of this complication. Influence of peritonitis on peritoneal transport and dialysis adequacy was also analyzed. There were neither persistent changes of peritoneal membrane permeability nor worsening of dialysis adequacy following peritonitis. PMID- 10835923 TI - [Diagnostic and therapeutic problems in primary antiphospholipid syndrome]. AB - The purpose of the paper is to estimate clinical problems of the primary antiphospholipid syndrome in a woman presenting recurrent fetal losses, thrombocytopenia, thrombo-embolic disorders, presence of lupus anticoagulant factor and high titer of antiphospholipid antibodies in serum. Clinical anamnesis, physical examination and laboratory findings including kidney biopsy excluded the possibility of SLE as a cause of this syndrome. In the perspective of the next pregnancy prophylaxis against pregnancy complication is discussed. PMID- 10835924 TI - [Abdominal hernia as risk for continuation of continuous ambulatory peritoneal dialysis: case report]. AB - On the case of 37 old patient with diabetic nephropathy, treated with peritoneal dialysis since 12 months, actual problems of clinical and imaging diagnostics of abdominal hernias in continuous ambulatory peritoneal dialysis patients are presented. We paid special attention to the postoperative management and prophylaxis of abdominal hernia recurrences. PMID- 10835925 TI - [Evaluation of peritoneal transport of solutes and water in patients treated with peritoneal analysis]. PMID- 10835926 TI - [Peritoneal transport of amino acids in continuous ambulatory peritoneal dialysis (CAPD)]. PMID- 10835927 TI - [Diet in chronic renal failure as the element of hygiene of life]. PMID- 10835928 TI - [Diagnostic and therapeutical progress in renal localization of plasma dyscrasias]. PMID- 10835929 TI - [Report of Seventh Assisi European Meeting on cardionephrology: April 15-17, 1999, Italy]. PMID- 10835930 TI - [Carotid artery disease in patients with retinal vein and artery occlusion]. AB - To evaluate carotid artery disease in patients with retinal vein occlusion (RVO) and with retinal artery occlusion (RAO), 41 RVO patients (male 21, female 20, mean age 63 +/- 12 years) and 59 RAO patients (male 39, female 20, mean age 66 +/ 12 years) were investigated. All patients were examined neurologically and underwent carotid ultrasound examination. Using carotid ultrasound, carotid artery disease was evaluated in terms of presence of plaque, echogenicity of the plaque, degree of stenosis, or presence of ulceration. Carotid plaque or occlusion of the carotid artery was observed more frequently in RAO patients than in RVO patients (ipsilateral side: p < 0.01, contralateral side: p < 0.001; Fisher's exact test). Heterogeneous plaque was found more frequently in RAO patients compared to RVO patients (ipsilateral side: p < 0.01, contralateral side: p < 0.02; Fisher's exact test). Ulcerated plaque was found only in patients with RAO. In conclusion, carotid artery disease was more frequently found in patients with RAO than in patients with RVO. PMID- 10835931 TI - [Respiratory function impairment in patients with Parkinson's disease--a consideration on the possible pathogenetic relation to autonomic dysfunction]. AB - To investigate the characteristics and clinical significance of respiratory function in patients with Parkinson's disease (PD), we studied 38 patients (male, 19; female, 19: mean age, 65.5 years: mean duration of disease, 6.7 years) who had no history of respiratory disease and smoking. Fifty three non-respiratory disease subjects (male, 26; female, 27: mean age, 67.6 years) were served as age matched control. We measured spirometry and maximal expiratory flow-volume curve in all patients, and analyzed the relations between respiratory function variables and clinical profiles. The clinical disability of PD was indicated by Hoehn-Yahr (H-Y) scale. The number of PD patients was 15 in H-Y 2, 18 in H-Y 3 and 5 in H-Y 4, respectively. The values of % VC, %FEV 1, FEV 1/FVC, %PEFR, % V50 in H-Y 4 group were significantly smaller than those in H-Y 2 and 3 groups. Small airway dysfunction (SAD) was represented by abnormality of % V25, % V50/V25. The prevalence of impairment in % V25 and % V50/V25 was detected in 13 patients (34.2%) and 15 patients (39.5%), respectively, this was significantly higher than age-matched controls. However, the mean value and prevalence of impairment in % V25, % V50/V25 were not affected by H-Y scale and duration of disease or ideal body weight (%predicted value). Twenty seven patients showed normal ventilatory function based on % VC over 80% and FEV 1/FVC over 70%. The prevalence of impairment in % V25, % V50/V25 was detected in 8 patients (29.6%), 9 patients (33.3%), respectively, among 27 patients with normal ventilatory function. These results suggest that ventilatory dysfunction is concerned with clinical disability but SAD which is independent of clinical disability seen prevalently in patients with PD. It is widely accepted that patients with PD frequently have cardiac or bowel dysfunction based on the visceral autonomic dysfunction. We hypothesize that SAD may also be caused by possible autonomic dysfunction in patients with PD. PMID- 10835932 TI - [A case of portal-systemic encephalopathy associated with Sheehan's syndrome]. AB - A 66-year-old woman had had recurrent episodes of disturbed consciousness whenever she had been constipated or dehydrated. She had been inactive and afflicted with obstinate constipation since she had menopause at age of 32. She underwent gastrectomy for gastric ulcer at age of 37. Laboratory examination showed marked hyperammonemia, reduction in Fisher ratio, and poor excretion of ICG. Furthermore, hypopituitarism and secondary hypothyroidism were found. She was diagnosed as Sheehan's syndrome. A T1-weighted MRI demonstrated symmetrical high intensity in the bilateral globus pallidus and empty sella. The histological examination of the liver revealed a mild lymphocytic infiltration without liver cirrhosis. Abdominal angiography showed a large shunt vessel between the splenic vein and the left renal vein. After embolization of the shunt vessel, hyperammonemia and neurological impairment improved. Additionally multiple hormones replacement has been useful to reduce the drugs of standard therapy for hepatic coma. In this case, we speculated that Sheehan's syndrome accelerated the constipation and worsened the shunt encephalopathy. PMID- 10835933 TI - [A patient with marked immunodeficiency in an HTLV-I carrier: a case report]. AB - We report a 49-year-old man who was an HTLV-I carrier with an immunodeficiency state and intracranial pyramidal tract lesion revealed by MRI. He was born in Hokkaido and was admitted to our hospital because of fluminant hepatitis. On admission, neurologic examination revealed exaggerated deep tendon reflexes including the jaw jerk; the plantar response was flexor. Laboratory examination revealed decrease in the number of lymphocytes and CD4-positive lymphocytes in the peripheral blood and CD4/CD8 ratio was consistently low, indicating the presence of cellular immunodeficiency state. Serum anti-HTLV-I antibody was markedly increased but he did not have HTLV-I associated myelopathy (HAM). He had no underlying disease which would cause immunodeficiency state such as adult T cell leukemia (ATL) or HIV infection. We concluded that the HTLV-I carrier state induced his immunodeficiency. During the course, he developed retrobulbar neuritis. T2 weighted cranial MRI revealed high signal lesions in the bilateral corona radiata, posterior limb of the internal capsule, and the pontine base, corresponding to the location of the pyramidal tracts. His hospital course was complicated by opportunistic infections such as Pneumocystis carinii pneumonia, cytomegalovirus infections, and meningitis, and died of multiple organ failure 7 months after the admission. Cellular immunodeficiencies in ATL patients are well known. Intracranial central nervous system (CNS) lesions in HAM patients are also mentioned. Recently coincidence of ATL and HAM in the same patients has also been reported. Asymptomatic HTLV-I carriers may have a latent immunodeficiency state and/or CNS lesions. We shall have to be alert about the presence of such carriers. PMID- 10835934 TI - [A case of polymyositis presenting pregnancy with acute respiratory failure]. AB - A 33-year-old pregnant woman developed respiratory difficulty with bilateral pleural effusion 31 weeks into gestation. On admission she had an elevated serum level of creatine kinase, but muscle weakness in the extremities was mild. After an immediate and successful Cesarean section, she developed respiratory failure and mechanical ventilation was required. The patient was diagnosed as having polymyositis from the limb muscle biopsy. She was treated with dexamethasone at a dose of 8 mg, methylprednisolone++, 1 g daily for three days, and then prednisolone 60 mg daily. One week later the serum level of creatine kinase was normalized and the patient was weaned from ventilator support. The dose of prednisolone was tapered 10 mg every week and she was discharged two months after delivery without prednisolone. A chest CT scan revealed no interstitial pneumonitis or aspiration pneumonia, so her respiratory failure seemed to be ascribable to polymyositis-related respiratory muscle weakness and pleural effusion. However, severe involvement of respiratory muscles without generalized marked muscle weakness is extremely rare, and pleural complications have usually been described in association with pulmonary parenchymal diseases. During the following two years polymyositis has not recurred in this patient and there have been no data indicative of other overlapping collagen diseases. Reports of polymyositis which occurred during pregnancy are rare, and the pathogenesis and clinical outcome of these patients need to be carefully investigated. PMID- 10835935 TI - [A case of sarcoid myopaty with external ocular muscle involvement--diagnosis and follow-up study with 99mTc pyrophosphate scintigraphy]. AB - A 62-year-old woman was admitted to our hospital because of diplopia and muscular weakness. She had been diagnosed as having sarcoidosis histologically by skin biopsy 5 years before. Although neither CT nor MRI could detect the granulomas, 99mTc-pyrophosphate scintigraphy successfully detected the involvement of sarcoidosis not only in the skeletal muscles but also in the external ocular muscles. After treatment with prednisolone, the symptoms markedly subsided and increased uptake of radionucleotide disappeared. Although 67Ga scintigraphy is also known to be useful in investigating the sites affected by sarcoidosis throughout the body, it was not effective in detecting the involvement of external ocular muscle due to the physiological uptake of 67Ga to the lacrimal grands. While the granulomas are frequently observed histologically in skeletal muscles, usually they are not associated with muscle symptoms. In our study 99mTc pyrophosphate scintigraphy has proven to be useful in investigating the nodular lesions in skeletal muscles of sarcoidosis than 67Ga scintigraphy. PMID- 10835936 TI - [Two families of Charcot-Marie-Tooth disease with Adie's pupil, axonal neuropahy and the Thr124Met mutation in the peripheral myelin protein zero gene]. AB - We reported two families of Charcot-Marie-Tooth disease (CMT) with Thr124Met mutation in the peripheral myelin protein zero (MPZ). The clinical features of the proband patients of both families showed Adie's pupil, severe sensory dominant neuropathy in lower extremities, and axonal changes in sural nerve biopsies and nerve conduction studies. Muscle atrophy and weakness was mild in the lower legs, while sensory impairment was marked. The proband patient of family 1 had four symptomatic siblings and one of them showed Adie's pupil. The elderly daughter of the proband of family 2 showed Adie's pupil and younger daughter showed photophobia. The biopsied sural nerves of both proband patients revealed prominent axonal sprouting, and sub-perineurial edema and mild fascicular enlargement. Segmental demyelination was not frequent in teased fiber assessment. The present two family cases strongly suggest that this MPZ gene mutation (Thr124Met) could be present among the patients with CMT type 2, axonal form. Furthermore, the patients showing sensory neuropathy and Adie's pupil may need to be reexamined with this mutation. It is also necessary to reassess genotype-phenotype correlation in CMT patients particularly in reference to type 1 and type 2. PMID- 10835938 TI - [A case of motor neuron disease with dementia, presenting motor aphasia as an initial symptom]. AB - We report a case of motor neuron disease (MND) with dementia, presenting motor aphasia as an initial symptom. A 67-year-old man was admitted to our hospital because of speech disturbance slowly progressing for 2 years. On physical examination, he showed no neurological abnormalities except for non-fluent aphasia and increased deep tendon reflexes without laterality. MRI demonstrated bilateral fronto-temporal atrophy, dominating the left hemisphere. This finding was confirmed by surface anatomy scanning (SAS), showing an obvious atrophy in the left inferior frontal gyrus, compared with the right one. SPECT with 123I-IMP revealed some irregular defects in the bilateral frontotemporal region. Because he showed dementia, bulbar palsy with tongue atrophy, weakness of upper extremities and facial muscles, snout reflex, and the atrophy and fasciculation in limbs in addition to motor aphasia soon after the discharge from our hospital, he was diagnosed as having MND with dementia. At age 68, he died of a respiratory failure 3 years after the onset of the disease. MND with dementia seldom shows motor aphasia as an initial symptom. We must include, however, the MND with dementia as an differential diagnosis when we see the patients with progressive aphasia. PMID- 10835937 TI - [Treatment with interferon-alpha 2a in a patient with chronic inflammatory demyelinating polyneuropathy]. AB - We report an adult patient with refractory chronic inflammatory demyelinating polyneuropathy (CIDP) who resisted conventional immunological therapies but could be successfully treated with interferon (IFN)-alpha 2a. A 56-year-old man was admitted to our hospital with progressive distal dominant muscle weakness and sensory disturbance of all extremities. He was diagnosed as CIDP based on laboratory examinations which revealed conduction block on peripheral nerve conduction studies and demyelinating findings on sural nerve biopsy. He was given several immunological treatments, including immunoadsorption therapy (IAT), oral corticosteroid, immunosuppressive agents such as cyclophosphamide and mizoribine, but without satisfactory efficacy although transient improvement of muscle weakness was observed for two weeks after IAT. A session of IAT every four weeks was therefore continued to maintain his symptoms for nine months. He was then treated with IFN-alpha 2a intramuscular injections every other day. Four weeks after starting of IFN-alpha 2a therapy, the worsening of his symptoms stopped and his muscle strength improved gradually to normal level without IAT, suggesting that IFN-alpha 2a may be effective in some patients with refractory CIDP. Further studies are needed to confirm the efficacy of IFN-alpha 2a for CIDP. PMID- 10835939 TI - [A case of Fisher syndrome showing pharyngeal-cervical-brachial weakness with an elevation of anti-GQ 1 b and anti-GT 1 a antibodies]. AB - A 15-year-old boy developed ataxic gait, diplopia and hoarseness. Within 3 days after the onset, he had additional symptoms of dysphagia and dysarthria. He was admitted to our hospital 7 days after the onset of the disease. On admission, he had total ophthalmoplegia, ataxia, areflexia, facial diplegia, bulbar palsy and weakness of the neck and upper arms. Serum anti-GQ 1 b and anti-GT 1 a antibodies were significantly elevated. A diagnosis of Fisher syndrome associated with pharyngeal-cervical-brachial weakness was made. He was placed on a high dose of intravenous immunoglobins (12.5 g/day x 2 days) and had steroid pulse therapy (methylprednisolone 1 g x 3 days), which resulted in an almost complete recovery. There have been no reports of Fisher syndrome associated with brachio-pharyngeal palsy. As in the case of the pharyngeal-cervical-brachial variant of Guillain Barre syndrome, anti-GT 1 a antibodies may be associated with Fisher syndrome with pharyngeal-cervical-brachial weakness. PMID- 10835940 TI - [A case of spinal muscular atrophy with marked calf hypertrophy and adolescent onset]. AB - We report on a 41-year-old male patient with spinal muscular atrophy (SMA). He had slowly progressive muscular weakness and hypertrophic calves since 14 years of age. The upper arms were slightly, and the thighs moderately atrophic, but the calves were remarkably hypertrophic. There was muscle weakness of both the upper and lower limbs, being more proximal in distribution. He had a positive Gowers' sign and his gait was slightly waddling. Serum creatine kinase level was elevated (518IU/l). Electromyogram revealed a neurogenic pattern. Muscle biopsy of the left biceps brachii showed chronic neurogenic changes. Immunohistochemical examination and Western blot analysis using anti-dystrophin antibodies showed no abnormality. DNA analysis with multiplex PCR proved no deletion in the dystrophin gene, while deletions of exons 7 and 8 of the telomeric copy of survival motor neuron gene were detected. In 1978, Pearn et al. described a new variant syndrome of SMA, characterized by adolescent onset, gross hypertrophy of calves, and a slowly progressive clinical course. The present case is compatible with this syndrome. Therefore, it is suggested that this syndrome, mimicking Becker muscular dystrophy, is not an independent clinical entity, although the phenotype of this syndrome is different from that of typical SMA. PMID- 10835941 TI - [A patient presented with atypical paroxysmal kinesigenic choreoathetosis and Becker muscular dystrophy]. AB - A 22-year-old man had choreatic movements in upper limbs, neck and trunk for over twelve years which were associated with dystonia in lower limbs upon initiating voluntary movements. The choreatic movement lasted for a few seconds and the dystonia lasted for a few minutes. He also had high serum CK levels and hypertrophic calf muscles. His muscle strength and deep tendon reflexes were normal. His choreatic movements fulfill the criteria for paroxysmal kinesigenic choreoathetosis (PKC). However, it was unclear what the symptom of dystonia was due to. From a muscle biopsy and DNA analysis, he was diagnosed as having Becker muscular dystrophy. Administration of anticonvulsant improved the dystonia as well as the choreatic movement, which showed that the dystonia was a symptom of PKC. Coincidence of choreatic movements and dystonias which had different lasting time in a patient of PKC was atypical and had not previously reported. PMID- 10835942 TI - [A case of HIV encephalopathy with dementia which showed significant improvement after the combination antiretroviral therapy]. AB - We report a 63-year-old male with HIV encephalopathy, whose initial symptom was acutely progressing dementia. He tested positive for HIV antibody, and HIV-RNA count was 2.8 x 10(5) copy/ml. All opportunistic infections that could cause dementia were ruled out. Dementia remarkably improved after the combination antiretroviral therapy of three types of drugs, two being nucleoside analog reverse transcriptase inhibitors and one being a protease inhibitor. The combination therapy could be very effective for the treatment of HIV dementia. PMID- 10835943 TI - [A case of steroid-resistant polymyositis who made a good response to high dose intravenous immunoglobulins (IVIG)]. AB - We report a 69-year-old woman with steroid-resistant polymyositis who made a good response to high dose intravenous immunoglobulins (IVIG). She was diagnosed as polymyositis by muscle weakness of her extremities and bulbar muscles, higher level of serum CK and muscle biopsy findings of her biceps brachii muscle. Because of the coexistence of pulmonary tuberculosis and interstitial pneumonia, she was initially treated with 40 mg (1 mg/kg)/day of oral prednisolone, and methylprednisolone pulse therapy 500 mg/day intravenous drip (d.i.v.) x 3 days, 750 mg/day d.i.v. x 3 days. Since her condition deteriorated in spite of these conventional therapies, IVIG therapy 15 g (375 mg/kg)/day d.i.v. x 3 days was administrated. She showed remarkable improvement of serum CK level and muscle strength. Although corticosteroid is the mainstay of therapy for polymyositis, our case study showed that IVIG is also an efficacious therapy for polymyositis, especially resistant to the conventional treatment. PMID- 10835944 TI - [A patient with choreoathetosis of the left upper extremity due to acute cerebral infarction]. AB - A 71-year-old woman was admitted to our hospital because of involuntary movement of the left upper extremity. MR image of the brain 15 days after the onset revealed the low intensity in right posterior limb of internal capsule. The lesion was surrounded by thalamus, subthalamic nucleus, and globus pallidus with enhancement by Gd-DTPA. Surface EMG revealed irregular grouped discharge in short duration and grouped discharge in long duration in the left upper extremity. Those features are compatible with one of choreoathetosis. Choreoathetosis due to cerebral infarction in acute phase is rare. We discussed pathophysiology of this involuntary movement due to lacunar infarction of posterior limb of internal capsule in acute phase. PMID- 10835945 TI - [Toxoplasmosis versus primary central nervous system lymphoma in AIDS]. PMID- 10835946 TI - We had better get moving! Join South Dakota Med PAC! PMID- 10835947 TI - Davison County I-3 pilot project for childhood immunizations in children under two years of age. AB - In 1995, a Davison County Immunization Subcommittee was organized to implement the Infant Immunization Initiative (I-3). The goal of the subcommittee was to improve the immunization rate in the county. Prior to the study, the immunization rate for Davison County was at 40% for children under the two years of age. Davison County had the lowest immunization rate in the state of South Dakota. At the conclusion of the study, 96% of children in Davison County were immunized by their second birthday according to recommendations of the Advisory Committee on Immunization Practices (ACIP), the American Academy of Family Physicians (AAFP), and the American Academy of Pediatrics (AAP). Davison County surpassed President Clinton's goal of 90% immunization rate for children under two years of age. PMID- 10835948 TI - TNK-tPA: a new thrombolytic for treatment of acute myocardial infarction. PMID- 10835949 TI - Facts and fallacies about varicella and the varicella vaccine. AB - Varicella is a nearly ubiquitous infection that results in significant morbidity and occasional mortality. The varicella vaccine has been licensed in the United States for the past five years. While varicella vaccination rates are slowly growing nationally, the immunization rate for South Dakota remains abysmally low. The legislature has recently mandated this vaccine for school entry in South Dakota, to promote its increased use. There are many fallacies about varicella and the varicella vaccine and questions about how new requirements of varicella vaccination before school entry will be met. This article identifies these common misconceptions and can serve as a resource for answers to questions commonly asked of vaccine providers. PMID- 10835950 TI - The new kid on the block. PMID- 10835951 TI - Systemic fungal infections in cancer patients. PMID- 10835952 TI - Probiotics and gastrointestinal function in health and disease. PMID- 10835953 TI - Clinical significance of hepatitis B markers. PMID- 10835954 TI - A multicentric evaluation of recombinant DNA hepatitis B vaccine of Cuban origin. AB - INTRODUCTION: Hepatitis B infection is an important public health problem all over the world. As no specific treatment is available, greatest emphasis is placed on prevention through immunization. OBJECTIVE: To evaluate the immunogenicity (sero-protection rate) and reactogenicity of a novel recombinant DNA hepatitis B vaccine of Cuban origin (Enivac HB). POPULATION AND METHODS: The study was conducted at two independent centres (New Delhi and Jaipur) and 111 healthy volunteers without any symptomatic or serological evidence of HBV infection were administered the vaccine intramuscularly in a dose of 20 ug at day 0, day 30 and day 180. Blood samples were collected for evaluation of sero protection on days 30, 60, 90 and 210 at both centres and, in addition, at day 120 at New Delhi. All subjects were evaluated for any adverse event for 120 hours subsequent to each dose of the vaccine. RESULTS: The mean age of the volunteers was 24 years and the male:female ratio was 61:50. An overall sero-protection rate of 24.3% was obtained at day 30, 68.5% at day 60, 94.5% at day 90 and 99.1% by day 120. Following the third dose of the vaccine (at day 180), we lost one subject to follow up and further evaluation could be done in a total of 110 volunteers. The sero-protection rate by day 210 was found to be 100% (110/110). The vaccine was well tolerated with no serious adverse reactions. Minor side effects such as injection site tenderness, erythema and/or low grade fever were observed in 4.5% of the subjects. CONCLUSIONS: The recombinant DNA hepatitis B vaccine of Cuban origin is safe, well tolerated and highly immunogenic. PMID- 10835955 TI - Comparison of polyethylene glycol versus combination of magnesium sulphate and bisacodyl for colon preparation. AB - Colon preparation using polyethylene glycol (PEGLEC) and combination of bisacodyl and magnesium sulphate was compared in 74 patients. Type of preparation did not influence patient discomfort. The quality of preparation was excellent with PEGLEC in 63.6% as compared to 41.6% with combination preparation. The completion rates were similar in both groups. Good preparation was linked with shorter procedure duration (p = 0.001) and greater depth of examination. No major side effects were noted with both preparations. To conclude, polyethylene glycol is recommended for screening of right colon while combination of bisacodyl and magnesium sulphate is a good and cheap preparation modality to screen the left colon. PMID- 10835956 TI - Histological comparison of chronic hepatitis B and C in an Indian population. AB - Many studies have shown that steatosis, lymphoid aggregates or follicles and bile duct injury on histology are more consistently associated with chronic hepatitis C than chronic hepatitis B. We compared liver biopsies of 30 patients of chronic hepatitis B with an equal number of age matched patients with chronic hepatitis C. Steatosis, lymphoid aggregates or follicles and bile duct injury were noted in 66.6%, 36.6% and 26.6% cases respectively of chronic hepatitis B as compared to 70%, 33.3% and 30% cases respectively of chronic hepatitis C. Thus none of the features were considered distinctive of HCV infection. PMID- 10835957 TI - Bio-artificial liver from cultured human foetal hepatocytes: feasibility and prospects. AB - BACKGROUND: Conventional therapy for acute liver failure has not been able to improve survival beyond 40%. Apart from liver transplantation, the most promising development in this field is the utilization of cultured hepatocytes to make 'bio artificial liver support systems' as a 'bridge to transplantation' or ideally as a 'bridge to total recovery'. This study examines the feasibility of culturing foetal hepatocytes without the use of growth factors and formulating a bio artificial liver support device in our set-up. METHODS: Foetal hepatocytes were harvested from the liver obtained from mid trimester abortions at Armed Forces Medical College and Command Hospital (SC), Pune. The liver was perfused with Phosphate Buffered Saline (PBS) and collagenase type IV and was cut with a pair of sterile scissors into tiny pieces. Cells so separated, were washed with PBS plus foetal calf serum and stirred to disperse the cell aggregates. Filtered cell suspensions were inoculated in polystyrene flasks containing hepatocyte culture medium (MEM E: 75%, M199: 25%, BSA: 0.1%, Bovine Insulin 5 micrograms/ml, FCS: 10%, Penicillin: 10 i.u., Streptomycin 50 micrograms/ml, Hydrocortisone 5 micrograms/ml and incubated at 37 degrees C. The functional capabilities of the cultured hepatocytes were analyzed by studying production of albumin and a foetoprotein. Structural integrity of hepatocytes was assessed by light and electron microscopy. RESULTS: The hepatocyte yield varied from 2 to 60 x 10(6) cells/L with an average of 38 x 10(6) cells/L in the eight consecutive experiments. Initial hepatocyte viability varied from 25% to 90% with an average of 61%. The yield and the viability of hepatocytes were adversely affected by the condition of foetus at birth and use of intra-amniotic injections for inducing abortions. Hepatocyte monolayers and colonies formed in 75% experiments. The cultures could be maintained in incubation without the use of epidermal or hepatocyte growth factors for 2-25 days with a mean survival of 8.9 days. The cells in culture were found to be structurally normal and functionally active and could be cryo-preserved. These hepatocytes were inoculated into a hollow fiber module to formulate bio-artificial liver support device. The cultures ultimately developed either cellular disintegration or bacterial infections despite use of antibiotics in the culture medium. CONCLUSIONS: We conclude that it is feasible to maintain foetal hepatocyte cultures without the use of expensive growth factors for over 8 days. Bio-artificial liver formulated with cultured foetal hepatocytes is now a step closer to clinical trials. PMID- 10835958 TI - Spontaneous splenic rupture secondary to metastatic gastric carcinoma. AB - Spontaneous rupture of the spleen secondary to metastatic carcinoma is rare. We report the second such case. Rupture is usually diagnosed at laparotomy. However ultrasound and computerized tomography can play a role in diagnosis. PMID- 10835959 TI - Amoebic liver abscess in pregnancy. PMID- 10835960 TI - Primary leiomyosarcoma of jejunal mesentery: a rare case. PMID- 10835961 TI - Turcot's syndrome in an Arab child. AB - A 12 year old female child of Arab origin presented with chronic bloody diarrhoea and growth failure. Physical examination confirmed a growth failure and cafe-au lait patches. Colonoscopy and colon biopsies showed adenocarcinoma of the colon and laparotomy confirmed metastases to the liver. A diagnosis of Turcot's Syndrome was entertained in this child. PMID- 10835962 TI - Enteropathogenic Escherichia coli isolates in paediatric diarrhoea. AB - Enteropathogenic Escherichia coli (EPEC) are known to cause infantile enteritis. We studied the prevalence of EPEC in paediatric patients with acute and persistent diarrhoea. A total of 56 stool samples from paediatric patients were studied. There were 28 significant bacterial isolates. Of these 21 were untypable E. coli, 5 were typable E. coli, four of which belonged to members considered to be enteropathogenic. Non E. coli isolates grown in pure culture were one each of Pseudomonas aeruginosa and Citrobacter freundi. The study reveals the definitive role of EPEC in childhood diarrhoea at all age groups and emphasizes the need for characterisation of all significant E. coli isolates in this age group. PMID- 10835963 TI - Benign oesophageal strictures in children of north India. PMID- 10835964 TI - Surgical audit: a must for speciality development in India. PMID- 10835965 TI - Association of Helicobacter pylori with peptic perforation in Chattisgarh region of India. AB - BACKGROUND: Helicobacter pylori is present in over 90% cases of peptic ulcer. There are very few reports regarding prevalence of H. pylori infection in peptic ulcer perforation which is the commonest complication of peptic ulcer disease. In the present study we have drawn this association. PATIENTS AND METHODS: 44 cases of peptic perforation were included in this study. Per-operative biopsy was taken from the margin of perforation. The criteria for H. pylori positivity were, Positive Rapid Urease Test and evidence of H. pylori infection on histological examination using Modified Giemsa and H&E stains. RESULTS: Prevalence of H. pylori infection in peptic perforation was 61.4%. This is well above the usual prevalence in normal population of 45%. H. pylori infection was significantly more common in cases of chronic dyspepsia (> 6 months) and not significantly related to sex, blood group, addiction, family history of acid peptic disease and previous H2 blocker therapy. INFERENCE: H. pylori is found more commonly in patients with peptic ulcer perforation than in those without. PMID- 10835966 TI - Two port laparoscopic cholecystectomy. PMID- 10835967 TI - [Magnetic resonance in choledocholithiasis]. PMID- 10835968 TI - [Prolonged cerebral disorders in the elderly after anesthesia and surgery]. PMID- 10835969 TI - [Oxidative stress and aging]. PMID- 10835971 TI - [Magnetic resonance imaging of patients with perianal fistulas]. AB - Magnetic resonance imaging (MRI) of the anal canal was retrospectively evaluated during a three year period in 67 patients with suspected perianal fistulae. In six cases the surgeon, who knew the results of the MRI, found a fistula not seen on MRI while MRI showed a fistula in one patient which could not be found at surgery. We recommend MRI in cases where an experienced surgeon has difficulties mapping the fistula tract. PMID- 10835970 TI - [MR-cholangiography in patients suspected with choledochus calculi]. AB - To test a new non-invasive imaging method, magnetic resonance cholangiography (MRCP), we compared MRCP with intraoperative cholangiography (IOC) or endoscopic retrograde cholangiography (ERC). In 55 patients with 11 common bile duct stones MRCP identified all 11 stones and incorrectly found stones in seven additional patients. Predictive value of positive test was 61%, predictive value of negative test was 100%. As a consequence of MRCP, ERC or IOC can be avoided in 67% of the patients. PMID- 10835972 TI - [Use of sedation, analgesia and neuromuscular blockade by intensive care units in Denmark 1996-1997]. AB - To assess the use of sedatives, analgesics and neuromuscular blocking agents in patients requiring mechanical ventilation in Danish ICUs, questionnaires were mailed to all Departments of Anaesthesiology in Denmark. Forty-nine out of 53 hospitals responded (92.5%). All ICUs surveyed used both sedatives and analgesics for patients undergoing mechanical ventilation. Opioids, benzodiazepines and propofol were employed most commonly, in particular by continuous infusion. Neuromuscular blocking agents were used at 65% of the ICUs surveyed, but only rarely. Ninety-eight percent of the ICUs reported side effects, but for less than 20% of the patients, secondary to the sedative treatment. Most frequent side effects were delayed emergence and gastrointestinal disorders. PMID- 10835973 TI - [Osteoid osteoma of the cervical spine]. AB - Osteoid osteoma of the cervical spine is a rare benign tumour. A case of a 26 year-old female with osteoid osteoma in C2 is presented. She had a six year history of pain in the neck region. She suffered from limited neck movement, accentuation of pain during the night, but distinctive salicylate pain relief. Scintigraphic bone scan and CT scan, rather than MRI-scan are the most useful for localisation of the tumour in the spine and are essential for optimal planning before surgery is performed. PMID- 10835974 TI - [Don't forget awareness anesthesia]. PMID- 10835975 TI - [Pregnancy and antibiotics]. PMID- 10835976 TI - [Giardiasis. Parasitologic diagnosis]. PMID- 10835977 TI - [C-reactive protein]. AB - C-reactive protein (CRP) is a reliable biochemical marker for tissue destruction, necrosis and inflammation. CRP is an essential human acute phase reactant produced in the liver as response to systemic stimuli. The biological half-life of CRP is not influenced by age, liver- or kidney function or pharmacotherapy. CRP values in acute bacterial infections have been appreciated for 70 years. The new standardized CRP analyses yield the possibilities of longitudinal monitoring of chronic inflammatory diseases and help identify complications. The more sensitive measures of the area of reference also supplies new information: As a prognostic marker for microvasculitis, CRP is at present re-writing the agenda for today's research in inflammation, angina pectoris, vascular insults and atherosclerosis. PMID- 10835978 TI - [C-reactive protein and infections in general practice]. AB - C-reactive protein (CRP) is a sensitive and non-specific inflammatory marker. The serum level of CRP starts to rise 6-12 hours after the start of an inflammatory stimulus. Sequential CRP measurements will have greater diagnostic value than a single measurement, and changes of the CRP values often reflect the clinical course. In use in general practice the diagnostic value of CRP is found to be high in adults with pneumonia, sinusitis and tonsillitis, however it is found to be low regarding otitis and pneumonia in children. As to urinary tract infections and salpingitis the value is still undefined. Measurement of CRP is an important diagnostic test but the analysis should not stand by itself but be evaluated together with the patient's history and clinical examination. PMID- 10835979 TI - [Air pollution by volatile organic compounds (VOC) and health complaints]. AB - BACKGROUND AND OBJECTIVE: Complaints of well-being and health after moving in a new or redeveloped building occur quite frequently. If these complaints persist for several months and are indicated by the majority of occupants, an examination of the air quality is required. However, the interpretation of these data can be difficult since different recommendations but no threshold limit values for the indoor contamination with volatile organic compounds (VOC's) exist. PATIENTS AND METHODS: This report presents a case with serious health complaints (14 males, average age 40.4 years; 44 females, average age 37.9 years) after moving in a completely redeveloped building. Complaints of well-being and health were investigated by a standardised questionnaire and compared with a control group (11 males, average age 41.4 years; 12 females, average age 33.3 years). The VOC contamination was analysed by various measurements. RESULTS: Two months after moving into the building a total VOC-concentration of 2000-3000 micrograms/m3 was registered, after 10 months the concentration decreased to 900-1300 micrograms/m3 due to intense airing. The following symptoms showed the clearest differences between exposed persons and the control group: soreness of throat (odds ratio: 10.72; 95%-confidence interval: 1.46-465.2), irritations of mucous membranes (OR: 10.45; 95%-CI: 1.43-453.8), headache (OR: 9.9; 95%-CI: 1.35-430.9) and increased weariness (OR: 7.55; 95%-CI: 1.55-71.2). As a consequence extensive redevelopment measures were initiated. CONCLUSION: Contamination of the indoor air with 900 micrograms/m3 VOC's (total value) can induce serious complaints of well-being and health. Rooms with VOC-concentrations > 1200-1500 micrograms/m3 are not suited for regular stay of unprotected people. PMID- 10835980 TI - [Echocardiographic measurements of coronary flow reserves in the left anterior descending artery allows detection of significant stenosis]. AB - BACKGROUND AND OBJECTIVE: CFR has proven to be useful in the selection of patients undergoing invasive treatment of coronary artery disease and in estimating their prognosis. However, CFR could only be determined in everyday practice invasively during catheterization procedures. Recent development of high resolution transthoracic color Doppler echocardiography (TTCD) allows transthoracic visualization of distal LAD and supra-apical intra-myocardial perforator branches, and non-invasive measurement of CFR. The feasibility of non invasive assessment of coronary flow reserve (CFR) in the left anterior descending artery (LAD) using echo-enhanced high resolution TTCD was investigated. The results were compared with the degree of coronary diameter stenosis obtained during cardiac catheterization. CFR was determined by measuring to ratio of pulsed wave Doppler time velocity integral during adenosine-induced hyperemia (140 micrograms/kg/min i.v.) to baseline value. If Doppler signal of LAD flow was insufficiently at basal condition, an echo enhancer (Levovist) was used. PATIENTS AND METHODS: 70 patients were examined by TTCD (7 MHz B-Mode, 5 MHz color Doppler, 3.5 MHz PW-Doppler) after coronary angiography had been performed. Gr I consisted of 14 patients without heart disease, Gr II of 26 patients with 40-75% isolated LAD diameter-stenosis, and Gr III of 30 patients with > 75% LAD diameter-stenosis. RESULTS: CFR could be quantified in 56/70 patients (80%), in 42/70 patients without echo enhancer, and in 14/28 patients with an echo-enhancing agent. CFR in Gr I was 3.84 +/- 0.57, in Gr II 2.31 +/- 0.20 (vs Gr I p < 0.01) and in Gr III 1.60 +/- 0.30 (vs Gr II p < 0.02). CONCLUSION: CFR of LAD can be determined in 80% of patients by the synergistic use of high resolution TTCD combined with intravenous given ultrasound echo enhancing agent. A coronary flow reserve of less than 2.1 detected in this patient cohort significant LAD-stenosis with a sensitivity and specificity of 91% and 76%. PMID- 10835981 TI - [Pruritus during treatment with an estrogen-gestagen combination]. AB - HISTORY AND CLINICAL FINDINGS: A 51 year old female patient complained about marked pruritus of the whole integument. Since seven years she had been taking a two phase combination of estrogens and gestagen because of climacteric symptoms. 24 years ago severe pruritus occurred especially at the hands during the last trimenon of her first pregnancy whereas 7 years later during the subsequent pregnancy pruritus was absent. Physical examination revealed multiple excoriations over the whole body. No other relevant pathological findings were noted. INVESTIGATIONS: Laboratory findings, chest X-ray and abdominal sonography were normal. TREATMENT AND COURSE: Neither oral administration of an antihistaminic drug nor topical application of betamethasone improved the situation to a relevant extent. For 12 months the occurrence of pruritus had not been referred to hormonal treatment by the physicians in charge. Complete cessation of pruritus was seen within a few days after the patient had stopped hormonal medication on her own. CONCLUSION: Patients should be asked for pregnancy associated pruritus or jaundice before receiving drug combinations with estrogens and gestagens. In case of a positive history, the physician should inform them about possible side effects such as pruritus and icterus. PMID- 10835982 TI - [Disseminated small-node cutaneous sarcoidosis]. AB - HISTORY AND ADMISSION FINDINGS: For 6 months before admission an 83-year-old woman had experienced itching from papules on her back. Despite local and systemic corticosteroid treatment the lesions had spread further over the back and also began to involve the proximal limbs. On examination the patient was noted to be fit for her age. At the described sites there were scattered red brown maculopapular lesions, pin-head sized, some circumscribed, others confluent. Diascopy demonstrated them to be greyish yellow. INVESTIGATIONS: Erythrocyte sedimentation rate was 15/42 mm, serum concentrations of angiotensin converting enzyme (ACE) and calcium were within normal limits. The Tine test was negative. Skin biopsy revealed a granulomatous epithelioid cell inflammation in the corium without central necrosis. She had diabetic retinopathy, a complication of type II diabetes mellitus for many years, and hepatic steatosis. TREATMENT AND COURSE: The results of the clinical and histological findings and exclusion of systemic involvement established the diagnosis of disseminated small node cutaneous sarcoidosis. Under clofazimine (Lampren), at an initial dosage of 3 x 100 mg daily by mouth, gradually reduced over 4 months, the cutaneous lesions healed completely and lastingly. CONCLUSION: This form of sarcoidosis has a good prognosis. Clofazimine, being efficacious and well tolerated, provides a good therapeutic alternative. PMID- 10835983 TI - [Diagnostic value of positron emission tomography (PET) in clinical oncology]. PMID- 10835984 TI - [Medical science in eastern Germany]. PMID- 10835985 TI - [The hard way to establish research culture in medicine]. PMID- 10835986 TI - [Administrative duty to inform at the completion of the selective payment agreement]. PMID- 10835987 TI - Pain assessment in infants and children. AB - The science of pain assessment for infants and children has grown substantially in the past several decades to the point that valid and reliable methods for pain assessment are available for use in clinical settings. Accurate pain assessment requires consideration of children's developmental level, type of pain experienced, history and context of pain, family influences, and interaction with the health care team. Research is needed to improve the sensitivity, specificity, and generalizability of pain-assessment tools and to more fully incorporate contextual factors into the objective assessment process. Finally, the improvement of pain assessment in the clinical setting can be viewed as a patient care quality issue, and continuous quality improvement methods can be used effectively to incorporate pain assessment as an integral component of every infant's and child's health care. PMID- 10835988 TI - Behavioral and cognitive interventions in the treatment of pain in children. AB - A wide range of behavioral and cognitive techniques have been found to be efficacious for helping children to cope with acute pain. Research into the clinical applicability of these interventions is at an early stage but increasingly is being recognized as an important future direction. Many existing interventions and assessment tools are reasonably easy to use, allowing practitioners to have the tools to identify children most vulnerable to pain and to significantly reduce pain-related distress in these children. For additional information from the Internet, please visit the UCLA Pediatric Pain Program Web site: http:?members.xoom.com/UCLAPAIN/. PMID- 10835989 TI - Weak analgesics and nonsteroidal anti-inflammatory agents in the management of children with acute pain. AB - The PSIs include acetaminophen, NSAIDs, and salicylates. They can be used alone for the treatment of mild pain or as an adjunct to opioid analgesia. In children, most experience is with acetaminophen and ibuprofen. For the treatment of mild to moderate pain, these agents can be administered as needed or at fixed intervals. The latter dosing scheme may provide a more consistent serum level, thereby improving analgesia. The major advantages of acetaminophen are its availability as a suppository for PR administration and its lack of effects on renal and GI function, adverse effects that may be seen with the NSAIDs. Many of the effects on platelet functioning, RBF, and the GI tract may be eliminated with the introduction of NSAIDs that selectively inhibit COX II without effects on COX I, the enzyme present in the GI tract, renal system, and platelets. Future evaluations with these agents in the pediatric population are needed. For more severe pain, the NSAID salicylate or acetaminophen can be combined with a weak opioid, such as codeine, oxycodone, or hydrocodone. When using oral analgesics, factors that may interfere with effective analgesia include a child's refusal to take the medication, ineffective doses and dosing regimens, decreased bioavailability following PO administration, inability to tolerate PO medications because of nausea or vomiting, altered GI motility, and a delayed onset caused by slow absorption. With such caveats in mind, the PO route provides an effective and cost-effective means for many patients. It should be considered as the primary route for pediatric patients in the treatment of mild to moderate pain, even in the hospital setting. PMID- 10835990 TI - Local anesthetics for the pediatric patient. AB - Local anesthetics are often extremely useful in the management of acute, chronic, and procedural pain. They have a low therapeutic ratio, so physicians should not exceed established dosing guidelines for a single bolus or long-term infusions. New local anesthetics that promise better safety and longer duration of action are on the horizon. PMID- 10835991 TI - Pediatric acute pain management. AB - The past decade has brought about an explosion of knowledge about the physiology of nociception and many new techniques for pain relief, new analgesic drugs, and new applications of old analgesic drugs. These techniques include methods of opioid administration by transdermal and transmucosal absorption and the use of neuraxial analgesia for the management of pain in children. Interest in the use of regional anesthesia in children has been rekindled, and analgesic properties and pre-emptive analgesic properties of many agents not typically considered analgesics, such as clonidine and ketamine, have been recognized. Perhaps the greatest advance has been the paradigm shift in the recognition that pain not only exists in infants and children but also is a significant cause of morbidity and even mortality. Given the unprecedented interest in pain management in adults and children, physicians can now look forward to the development of new methods of drug delivery and of receptor-specific drugs that divorce analgesia from the untoward side effects of existing analgesics. Improvement in the quality of life of hospitalized children also will occur. PMID- 10835992 TI - Complementary therapies for acute pediatric pain management. AB - A wide variety of tools to adequately treat pediatric pain is beneficial. The methods discussed herein typically involve the use of many areas of expertise to manage pain. Massage therapists, biofeedback technicians, physician acupuncturists, child-life specialists, psychologists, and physical or occupational therapists can all be used as allies to battle acute pain in children. The incorporation of alternative forms of pain management, including education, relaxation techniques, hypnosis, guided imagery, biofeedback, and even acupuncture, to the standard methods may improve the management of children with acute pain. The management of children with pain does not have to be with an "either/or" approach using traditional pharmacologic methods or the cognitive and alternative therapies discussed here. Many areas need research to provide evidence that these therapies work well. What is known now suggests that the use of these adjunctive methods of pain management may complement pharmacologic pain management, thereby bringing physicians closer to optimal care of children with acute pain. PMID- 10835993 TI - Office-based pain management. The 15-minute consultation. AB - Pain is a ubiquitous component of pediatric illness and injury. Unfortunately, the treatment of pain has remained secondary to the diagnosis and treatment of the disease state. Clinicians' duty is to relieve suffering, and, as such, the relief of pain should be a primary focus. Presently available modalities, when used appropriately, can significantly reduce the pain associated with medical encounters. Careful attention to such approaches may reduce the anxiety about pain, which is often an undercurrent at most health care visits, and allows clinicians and children to focus on getting well and staying healthy. PMID- 10835994 TI - Migraine and headache in childhood and adolescence. AB - Headache is one of the most common physical complaints of children and adults. The authors have provided definitions of headache, a classification system, diagnostic evaluations appropriate for children, and treatment options for patients with acute and chronic headache. Also, this article has emphasized the importance of diagnosing and treating migraine headache, a painful malady that is extensively underestimated and misdiagnosed in the pediatric population and one that can be treated acutely and when appropriate prophylactically with great success. Lack of a specific biologic marker, specific investigation, or brain imaging reduce these clinical entities too often to a psychological illness. Nonpharmacologic treatments are pivotal to manage chronic headaches. Migraine therapy, if administered early and through the appropriate route, could provide important and rapid relief. PMID- 10835995 TI - Treatment of pain in the neonatal intensive care unit. AB - Pain is a disruptive influence on infants in the NICU. The most obvious and effective strategy to decrease infant pain in the NICU is to stringently limit the frequency of painful procedures, especially those that are most commonly reported (i.e., heel lances and endotracheal suctioning), and have these performed on infants that are most unstable or critically ill by the most experienced person available. Organizational concerns over the cost of NICU care have forced a re-evaluation of the necessity of certain diagnostic and care procedures and a limiting of procedures to those that can be documented to positively affect clinical outcome. Pharmacologic and nonpharmacologic strategies are essential to the prevention and management of neonatal pain, and these should be considered for complementary use for every infant. Research has shown the safety and effectiveness of some of the strategies in reducing mild pain caused by brief invasive procedures; however, many of the studies had methodologic limitations. Therefore, more research is required to determine the comparative efficacy of the various strategies and to document additive or synergistic effects when combined. PMID- 10835996 TI - The management of pain in the emergency department. AB - The challenge for emergency medicine physicians in the new millennium is to use these drugs and drug combinations to make ED visits pain-free and safe experiences. With dedication to research, a willingness to take the time to explore new options, and expansion of pharmacologic and nonpharmacologic interventions, physicians can make this lofty dream a reality. PMID- 10835997 TI - Burn pain management in children. AB - Burn injuries present complex management problems that are exacerbated by the pain and suffering associated with burns. This pain commands primary consideration in the treatment plan, and the goal should be to achieve the least pain possible. This article describes strategies for assessing children's pain and for treating pain with a combination of pharmacologic and complementary therapies. An experienced interdisciplinary team is essential to successful pain management. PMID- 10835998 TI - The management of pain in sickle cell disease. AB - The pain of vaso-occlusive crisis in patients with sickle cell disease is excruciating, incapacitating, and sometimes refractory to even the most advanced analgesic treatments. A comprehensive, multimodal approach to therapy that includes education, cognitive therapies, anti-inflammatory drugs, opioids, and psychostimulant adjuvant drugs has been presented. Until a cure for the underlying disease is found, these are the best approaches available. The authors hope that future research will find even better modalities of analgesic care. PMID- 10835999 TI - Pain and symptom control in terminally ill children. AB - The management of pain in terminally ill pediatric patients has incalculable benefits to patients, their families, and physicians and nurses. A therapeutic management plan is dependent on a thorough understanding of the causes of pain in these patients, on pain assessment, and on the myriad drugs and drug strategies that are essential in pain treatment. Aggressive symptom control of treatment related side effects can ensure successful implementation of such a plan. PMID- 10836000 TI - Molecular basis of cancer and clinical applications. AB - This article attempts to show the vertiginous advances that exist today in the concept of what cancer is. The authors chose some multiple biologic concepts that have enabled the progress in the knowledge of this disease to occur at a speed no one could imagine until recently. Although the areas and biologic problems that remain to be solved are more numerous and complex than they expected, the basic fundamentals already partially understood and the multidisciplinary integration of the various medical specialties with biomolecular research enable physicians to face the next millennium with great optimism about the possibilities of therapeutic success, prevention, and effective early diagnosis. PMID- 10836001 TI - The changing role of the surgical oncologist. AB - It is increasingly important for the surgical oncologist and surgeons to have a thorough understanding of the advantages and limitations of adjuvant systemic chemotherapy, hormonal therapy, and adjuvant radiotherapy in various resectable cancers. Justification for the field of surgical oncology should include the fact that enough knowledge has been acquired about these adjunctive treatments for patients with cancer that they can be integrated into overall management. Too often in the recent past, surgeons, after the technical surgical resection, turned over the entire management of patients to the medical oncologist or the radiotherapist. Comprehensive management for surgeons and surgical oncologists should maintain their voice in management policy so that the patients can be served best by the application of adjuvant treatment or by the avoidance of adjuvant treatment, systemic or local, when it is not appropriate or significantly helpful. The overall justification for the field of surgical oncology is that the surgeon not only knows how to use medical treatments in advanced cancers, but also knows how to select minimal surgical procedures in early cancers and how to manage and direct the application of adjuvant treatments, regionally and systemically. PMID- 10836002 TI - Outcome assessment in cancer management. AB - The goals of cancer treatment may be prevention, cure, or palliation. In each case, the length and quality of survival are the ultimate outcomes to assess when evaluating the success of treatment. Some surrogate or intermediate endpoints, however, may be considered, such as toxicity or the tumor response. The first section of this article considers traditional outcomes and endpoints and intermediate or surrogate endpoints for survival and quality of life. The second section specifically addresses the issue of the quality-of-life outcome, which has become increasingly important over the last 2 decades. The last section suggests that combining survival and quality of life may not be easy. Costs are increasingly important in management of diseases, and the outcomes of cancer treatment frequently are measured against them. Although the definitions of cost effectiveness and cost-utility are mentioned at the end of this article, the article does not deal with the problems of decision making for health resource allocation. PMID- 10836003 TI - Reading and assessing reports of treatment studies in oncology. AB - Rapid advances in tumor biology, immunology, genomics, and technology give physicians great hopes for providing patients with better chances in the struggle against cancer. The pace of progress will be slowed, however, if we do not have clear answers regarding which treatments work and do not work. Such answers come from carefully designed, randomized, clinical trials. Such trials require infrastructure, commitment, cooperation, time, and money, and they provide little fame. They are, however, an invaluable contribution of the medical profession to their patients, to their next generation of colleagues, and to future patients. Randomized clinical trials that answer important medical questions definitively should be supported, participated in, and demanded by surgeons and oncologists. PMID- 10836004 TI - Role of videoscopic-assisted techniques in staging malignant diseases. AB - Preoperative radiographic studies fail to uncover a significant number of patients with unresectable malignancies. Small peritoneal studs of tumor, lymph node involvement, and small liver metastases are common causes of understaging by imaging studies. Videoscopic staging offers a higher degree of accuracy among these patients, with minimal complications and a shorter recovery time than with exploratory laparotomy. The addition of laparoscopic sonography and peritoneal lavage may further increase sensitivity to unresectable disease. Staging videoscopy has been applied to hepatobiliary and gastrointestinal malignancies, pleural and pulmonary tumors, and gynecologic malignancies. The author believes that videoscopic staging will become increasingly common as it is further studied and the best applications are delineated. PMID- 10836005 TI - Multidisciplinary care for patients with breast cancer. AB - Breast cancer management requires a multidisciplinary approach that is tailored to the patient's stage at presentation, desire for breast conservation or reconstruction, estimation of risk of recurrence, and assessment of the benefits and toxicities of potential adjuvant therapies. At the Lahey Clinic Medical Center, breast surgeons, plastic surgeons, radiation oncologists, and medical oncologists staff the Breast Cancer Treatment Clinic, and work closely together to formulate treatment plans that will optimize the likelihood for cure with an acceptable cosmetic result. This involves careful preoperative work-up, surgical axillary staging, breast irradiation in the setting of breast conservation, and selection of chemotherapy or hormonal therapy if appropriate. Newer aspects of breast cancer care, including sentinal lymph node biopsy, postmastectomy radiation therapy, expanded use of hormonal therapy in younger women, new agents and chemotherapy combinations, and autogenous reconstruction techniques, have become an essential part of the multidisciplinary clinic approach. PMID- 10836006 TI - Treatment of colon and rectal cancer. AB - This article discusses multimodal treatment of noncomplicated colon and rectal cancer, considerations for specific types of colon cancer, considerations that may modify the extent and technique of surgery, the role of adjuvant chemotherapy for colon adenocarcinoma and rectal cancer, and surgical treatment of complicated colorectal cancer. PMID- 10836007 TI - Thyroid cancer. AB - The management of patients with thyroid cancer can be optimized by developing multidisciplinary groups of highly specialized individuals. The completeness of surgery and its morbidity are mostly surgeon-dependent. Similarly, the decisions regarding selection of adjuvant treatments, doses, follow-up schemes, and so forth require depth of knowledge and understanding of the disease; its variables; factors that govern its course; and the values, limitations, and side effects of alternative therapies. PMID- 10836008 TI - Melanoma. A multidisciplinary approach for the general surgeon. AB - Advances in the understanding of the biology and treatment of melanoma have moved the care of melanoma patients into an increasingly multidisciplinary environment. Surgeons must understand these advances because they will often be responsible for directing the overall care of these patients. Most patients with melanomas more than 1 mm in diameter and no evidence of metastatic disease should be offered SLNB to more accurately stage them and direct decisions about participation in postoperative adjuvant therapy trials. Until the results of the MSLT are known, the effect of SLNB and ELND on outcome remains unknown. SLNs should be analyzed with serial sectioning and immunohistochemistry to avoid missing micro-metastatic disease. Based on the results of the ECOG-1684 trial, the FDA approved IFN-alpha 2b for the adjuvant treatment of melanoma patients with thick primary tumors (> 4 mm) or resected nodal disease. IFN-alpha 2b treatment is expensive and potentially toxic. The data from ECOG-1684 do not support the use of IFN-alpha 2b in patients with node-negative disease. In light of the ECOG-1690 trial results, the role of high-dose IFN-alpha 2b in the management of patients with resected nodal disease is considerably less clear. Any recommendations for treatment with high-dose IFN-alpha 2b should be made only after weighing the costs, side effects, and potential benefits for individual patients. Numerous, less toxic, promising, adjuvant immunotherapeutic strategies have been developed and are being tested in multicenter, prospective, randomized trials. These strategies include GMK, PMCV, and Melacine. If the results of any of these trials show a survival advantage compared with placebo or equivalent survival compared with IFN-alpha 2b, these immunotherapeutic agents will become the adjuvant treatment of choice for patients with resected high-risk melanoma. RT-PCR detection of tyrosinase in SLNs can identify patients with submicroscopic nodal disease who may be at increased risk for recurrence or death from melanoma. An ongoing, prospective, randomized trial will determine whether patients with histologically negative but RT-PCR-positive SLNs will benefit from lymphadenectomy or adjuvant IFN-alpha 2b therapy. RT-PCR can also identify minimal residual disease in peripheral blood and bone marrow from patients with high-risk melanoma, but RT-PCR analysis of peripheral blood and bone marrow is still experimental, and procedural details need to be standardized and prospectively validated in large patient groups before its use can be considered the standard of care. PMID- 10836009 TI - Hepatic malignancies. AB - The battle against malignancies of the liver is far from over, although tremendous strides have been made in the past decade, such as improved diagnostic capabilities, safe surgical resection, availability of safe nonsurgical ablative modalities, multimodality therapy, and aggressive approach to recurrent disease. Even after the best attempts at curative treatment, recurrence of primary and secondary malignancies of the liver continues to be the cause of demise for more than 70% of treated patients. The battle continues in the laboratories, where investigations are focused on delineating the pathophysiology of cancer on the molecular and genetic levels and mapping the patterns of cancer emergence and spread. The new millennium holds promise for formulating therapies that may improve disease-free survival for patients with malignancies of the liver. PMID- 10836010 TI - Secondary pulmonary malignancy. AB - Patients with pulmonary metastases were previously relegated to palliative medical management. Since the first metastasectomies in the nineteenth century, general acceptance of this technique has occurred. Although, initially, indications for resection of pulmonary metastases were limited to patients with solitary nodules, over time, indications have broadened to include multiple lesions, recurrent disease, and nearly all histologies. With appropriate patient selection and the absence of extrathoracic disease, survival may be improved. For patients with disseminated and symptomatic disease, surgical therapy may also provide some relief. PMID- 10836011 TI - Multidisciplinary approach to esophageal and gastric cancer. AB - Despite marked advances in surgical therapy for patients with esophageal, esophagogastric, and gastric cancers, the overall prognosis of these patients has not markedly improved during the past decades. Multidisciplinary approaches using adjuvant postoperative and neoadjuvant preoperative therapeutic principles have received increasing attention with regard to the management of these patients. A series of randomized, prospective trials has demonstrated that adjuvant postoperative radiation or chemotherapy does not result in a convincing survival advantage after complete tumor resection in esophageal, esophagogastric junction, or gastric cancer. The available data on the role of neoadjuvant preoperative therapy are not yet conclusive. Although neoadjuvant therapy may reduce the tumor mass in many patients, several randomized, controlled trials have shown that, compared with primary resection, a multimodal approach does not result in a survival benefit in patients with locoregional, that is, potentially resectable, tumors. In contrast, in patients with locally advanced tumors, that is, patients in whom complete tumor removal with primary surgery seems unlikely, neoadjuvant therapy increases the likelihood of complete tumor resection on subsequent surgery, but only patients with objective histopathologic response to preoperative therapy seem to benefit from this approach. Consequently, in the future, improvements in the overall survival of patients with esophageal, esophagogastric junction, or gastric cancer most likely will be achieved only by tailored therapeutic strategies that are based on the individual tumor location, tumor stage, and consideration of established prognostic factors. A clear classification of the underlying tumor entity, a profound knowledge of the prognostic factors applicable, a thorough preoperative staging, and identification of parameters that allow for the prediction of response to preoperative therapy will become essential for the selection of the optimal therapeutic modality for individual patients. PMID- 10836012 TI - Soft-tissue sarcomas. AB - Sarcomas of the soft tissues are challenging lesions for the surgical oncologist. Careful planning must be done at all stages of diagnosis and treatment, because every sarcoma is unique with respect to histologic type, size, and location. Pretreatment discussions in a multidisciplinary format are useful to ensure appropriate and effective management of these tumors. PMID- 10836013 TI - Multidisciplinary considerations for patients with cancer of the pancreas or biliary tract. AB - The past century has been nearly all of the growth in knowledge about the anatomy and pathophysiology associated with cancers of the pancreas and surrounding biliary structures. Through advances in imaging technology, endoscopic practice, improvement in surgical technique and perioperative care, anesthesia advances, and a better appreciation for the usefulness of adjuvant chemotherapy and radiation therapy, physicians can offer patients some hope for long-term survival and a better quality of life when they are faced with these devastating tumors. Although surgical intervention is the "last best hope" for these patients, advances in the nonoperative disciplines will be required for substantial further improvement in patient outcomes. PMID- 10836014 TI - Multidisciplinary care of the terminally ill patient. AB - This article discusses sedation, the assessment and management of physical symptoms, and symptom-assessment scales for the terminally ill patient. The evaluation of the ability of the family or community to care for a terminally ill patient in pain also is discussed. PMID- 10836015 TI - Childhood tumors. AB - Pediatric solid tumors represent a distinct set of malignancies of embryonal origin whose incidence peaks in the first years of life. Specific genetic anomalies with pathogenic significance, which have helped to define the diagnosis better and to improve the prognosis of children with these tumors, recently have been discovered. Survival of children with solid tumors also has improved significantly because of effective multidisciplinary care, which, in this case, always involves chemotherapy and surgery. These favorable results require that children with these diseases are referred and treated at institutions that have multidisciplinary teams and the infrastructure and expertise for caring for these children. Diagnostic and therapeutic principles for the most common childhood solid tumors are discussed in this article, with an emphasis on surgical procedures. PMID- 10836017 TI - Aminoethylprolyl peptide nucleic acids (aepPNA): chiral PNA analogues that form highly stable DNA:aepPNA2 triplexes. AB - [formula: see text] The replacement of the glycyl component in the peptide nucleic acid (PNA) backbone by a prolyl unit bearing a nucleobase leads to the aminoethylprolyl (aep) PNAs, which are chiral and cationic. The homooligomeric aepPNA binds to complementary DNA sequences with high affinity and sequence specificity, forming highly stable triplexes. PMID- 10836016 TI - Significance and management of local recurrences and limited metastatic disease in the abdomen. AB - The management of patients with synchronous or metachronous metastatic carcinoma, sarcoma, or melanoma in the abdomen requires a knowledge of the natural history of the disease and of the available treatment options. Patients with advanced malignant disease may be of marginal performance status yet may require large surgical procedures or combined modality therapy; the most challenging therapeutic decisions involve such patients. The authors highlight the role of surgery in selected patients with metastatic or recurrent malignancy as it is practiced at The University of Texas M. D. Anderson Cancer Center. PMID- 10836018 TI - Efficient synthesis of 2-deoxy L-ribose from L-arabinose: mechanistic information on the 1,2-acyloxy shift in alkyl radicals. AB - [formula: see text] Conversion of the inexpensive L-arabinose 1 into the ethylthio ortho ester 7 followed by generation of the dialkoxyalkyl radical III produces the desired 2-deoxy-L-ribose triester 4 in excellent overall yield. It has been shown that the similar dialkoxyalkyl radical IV is not an intermediate in the 1,2-acyloxy shift of anomeric radical I. PMID- 10836019 TI - Promiscuous Watson-Crick cross-pairing within the family of pentopyranosyl (4'- >2') oligonucleotides. AB - [formula: see text] The D-beta-ribo, D-beta-xylo, L-alpha-lyxo, and L-alpha arabino members of the pentopyranosyl (4'-->2') oligonucleotide family show efficient intersystem cross-pairing among each other. This family of configurationally isomeric and conformationally well-defined pairing systems offers an opportunity to study structural factors that determine cross communication between informational oligonucleotide systems of different backbone structure. PMID- 10836020 TI - L-alpha-lyxopyranosyl (4'-->3') oligonucleotides: a base-pairing system containing a shortened backbone. AB - [formula: see text] The L-alpha-lyxopyranosyl (4'-->3') oligonucleotide system shows cooperative base-pairing in spite of containing only five instead of the usual six covalent bonds per repetitive backbone unit. In contrast, corresponding D-beta-ribofuranosyl (4'-->3') oligonucleotides do not show adenine-thymine pairing under comparable conditions. The difference in pairing behavior relates to the conformation of the two systems' vicinal 3',4'-phosphodiester substituents, which is diaxial in the lyxopyranosyl system and 3'-axial-4' equatorial in the ribopyranosyl system. PMID- 10836021 TI - Studies on inducers of nerve growth factor: synthesis of the cyathin core. AB - [formula: see text] Compounds that induce the synthesis of nerve growth factor (NGF) are of interest as alternatives to the administration of the native peptide. We have initiated a program to study the NGF synthesis stimulating activity of the erinacine and scabronine diterpenes. Herein, we report an approach to the core cyathin system by sequential application of an oxidative coupling and [4 + 3] cycloaddition. PMID- 10836022 TI - beta-Siloxy unsaturated nitriles: stereoselective cyclizations to cis- and trans decalins. AB - [formula: see text] beta-Siloxy unsaturated nitriles are excellent precursors to enolate and nitrile anions that cyclize to cis- and trans-decalins, respectively. The stereoelectronic requirements of endocyclic enolates and exocyclic nitrile anions are complementary, providing cis- and trans-decalins from a common intermediate. This cyclization allows the synthesis of diastereomeric cis- and trans-decalins containing a contiguous array of quaternary-tertiary-quaternary stereocenters. PMID- 10836023 TI - Methylsulfenylation of thioacetals as a method for synthesizing 2-thio substituted furans. AB - [formula: see text] The dimethyl(methylthio)sulfonium tetrafluoroborate induced cyclization of various bis(methylsulfanyl) carbonyl compounds is described. The reaction proceeds by methylthiolation of the thioacetal group to give a thionium ion which undergoes subsequent cyclization with the neighboring carbonyl group. This is followed by an elimination reaction to furnish the furan ring. PMID- 10836024 TI - Enantioselective ring opening of meso aziridines catalyzed by Tridentate Schiff base chromium(III) complexes. AB - [formula: see text] A catalytic method for the enantioselective ring opening of meso aziridines by TMSN3 is described. Tridentate Schiff base chromium complexes derived from 1-amino-2-indanol were identified as the optimal catalysts. PMID- 10836025 TI - Sets of aldolase antibodies with antipodal reactivities. Formal synthesis of epothilone E by large-scale antibody-catalyzed resolution of thiazole aldol. AB - [formula: see text] Three monoclonal aldolase antibodies, generated against a beta-diketone hapten by reactive immunization, catalyzed rapid and highly enantioselective retro-aldol reactions of ent-8a-k, providing optically pure 8a-k by kinetic resolution. Compounds (+/-)-8a, (+/-)-8g, and (+/-)-8k have been resolved in multigram quantities using 0.003, 0.005, and 0.0004 mol% antibody catalysts, respectively. Resolved compounds 8a-k are useful synthons for the construction of epothilones A-E (2-6) and their analogues. Here, a formal synthesis of epothilone E, 6 has been achieved starting from compound 8g. PMID- 10836026 TI - Nitroazole universal bases in peptide nucleic acids. AB - [formula: see text] The syntheses of PNA oligomers containing potential ambiguous nucleobase analogues, namely 3-nitropyrrole and 5-nitroindole, have been accomplished. Hybridization properties of these PNAs with complementary oligodeoxynucleotides were evaluated by thermal denaturation experiments. Both novel residues exhibited little variation in Tm (< or = 1.5 degrees C) when positioned against any of the four nucleoside bases. The capability to incorporate degenerate sites should further expand the utility of PNA in applications where precise sequence information is not available. PMID- 10836027 TI - Facile synthesis of substituted phenanthroline ligands by samarium-promoted coupling of phenanthroline with ketones. AB - [formula: see text] 1,10-Phenanthroline undergoes coupling with ketones promoted by samarium diiodide to produce 2-(1-hydroxyalkyl)-1,10-phenanthrolines. O Methylation of these derivatives provides the corresponding 2-(1 methoxyalkyl)phenanthrolines. Demethoxylation with samarium diiodide then affords 2-alkylphenanthrolines. This process may be repeated to obtain 2,9-disubstituted phenanthrolines. A variety of new, substituted phenanthrolines are thus obtained. These compounds have numerous potential applications as ligands in metal-promoted reactions, including asymmetric catalysis. PMID- 10836028 TI - First total synthesis of xestobergsterol A and active structural analogues of the xestobergsterols. AB - [formula: see text] A novel pentacyclic polyhydroxylated sterol, xestobergsterol A (1a), has been synthesized in 24 steps and in good overall yield from stigmasterol 17. The key steps of the synthesis are the Breslow remote functionalization of the polyoxygenated steroid derived from 25 and the base catalyzed epimerization-aldol condensation of the dione derived from 27. PMID- 10836029 TI - Synthesis of a novel chiral binaphthyl phospholane and its application in the highly enantioselective hydrogenation of enamides. AB - [formula: see text] A new chiral phosphine, (R,R)-1,2-bis[(R)-4,5-dihydro-3H dinaphtho[2,1- c:1',2'-e]phosphepino]benzene [abbreviated as (R,R)-binaphane] was prepared on the basis of a practical route from a readily accessible enantiomerically pure binaphthanol. This ligand possesses both binaphthyl chirality and phospholane functionality. Excellent enantioselectivities (95-99.6% ee) have been observed in hydrogenation of an isomeric mixture of (E)- and (Z) beta-substituted-alpha-arylenamides by using a Rh-binaphane catalyst. These enantioselectivities are the highest reported to date for this transformation. PMID- 10836030 TI - Non-hydrogen-bonded secondary structure in beta-peptides: evidence from circular dichroism of (S)-pyrrolidine-3-carboxylic acid oligomers and (S)-nipecotic acid oligomers. AB - [formula: see text] Homooligomers of beta-amino acids (S)-3-pyrrolidine-3 carboxylic acid (PCA) and (S)-nipecotic acid (Nip) were studied by circular dichroism (CD) in methanol. In each series, a profound change in the far-UV CD spectrum was observed from monomer to tetramer, but little change was observed from tetramer to hexamer. A comparable pattern is observed in the CD spectra of short proline oligomers. We conclude that both PCA and Nip oligomers with > or = four residues adopt a characteristic secondary structure. PMID- 10836031 TI - Diaryl ethers using Fischer chromium carbene mediated benzannulation. AB - [formula: see text] The biological relevance and irresistible synthetic challenge of compounds containing the diaryl ether linkage encourages the development of new methodologies targeted toward this structural subunit. The syntheses of diaryl ethers 2 using a benzannulation strategy that formally involves a [3 + 2 + 1] cycloaddition between aryloxy-substituted Fischer carbenes 1 and alkynes are described. This methodology provides a neutral near ambient temperature formation of diaryl ethers. PMID- 10836032 TI - Solid-phase enzymatic synthesis of oligonucleotides. AB - [formula: see text] The controlled and selective synthesis of oligonucleotides on the solid phase is possible under mild aqueous conditions using the enzyme T4 RNA ligase, the resins being tentagel or kieselguhr/polydimethylacrylamide. PMID- 10836033 TI - Epicalyxin F and calyxin I: two novel antiproliferative diarylheptanoids from the seeds of Alpinia blepharocalyx. AB - [formula: see text] Epicalyxin F (1) and calyxin I (2), two novel diarylheptanoids, were isolated from a residual fraction of an EtOH extract of Alpinia blepharocalyx. Calyxin I (2) represented a new carbon skeleton, and epicalyxin F (1) possessed potent antiproliferative activity toward HT-1080 fibrosarcoma and colon 26-L5 carcinoma with ED50 values of 1.71 and 0.89 microM, respectively. PMID- 10836034 TI - A practical new chiral controller for asymmetric Diels-Alder and alkylation reactions. AB - [formula: see text] The enantiomerically pure hydroxy sulfones (+)- and (-)-2 have been prepared from 1,2-epoxycyclohexane by a simple and practical procedure. The acrylate esters of these alcohols undergo BCl3-catalyzed Diels-Alder reactions with a variety of dienes at -78 to -55 degrees C in CH2Cl2 or C7H8 with high dienophile face selectivity (Table 1). The chiral esters so formed are readily cleaved with recovery of the controllers (+)- or (-)-2. Esters of (+)- and (-)-2 can be converted to Z-potassium enolates and alkylated with high face selectivity. PMID- 10836035 TI - A biomimetic synthesis of the pauciflorine A and B skeleton. AB - [formula: see text] Racemic minovincine (3), following cyclization, reduction, O tosylation, fragmentation, and carbamate formation, provided the deoxypauciflorine 12. PMID- 10836036 TI - Synthesis of trisubstituted alkenes via olefin cross-metathesis. AB - [formula: see text] Trisubstituted alkenes have been prepared for the first time via intermolecular olefin cross-metathesis, using 1,3-dimesityl-4,5 dihydroimidazol-2-ylidene ruthenium alkylidene complexes 3a,b in good yields with moderate E selectivity. In addition, protected alcohols near the geminal disubstituted olefin improve reactivity for cross-metathesis. PMID- 10836037 TI - Solvent-dependent stereoselectivity of bis-2-pyridone [4 + 4] photocycloaddition is due to H-bonded dimers. AB - [formula: see text] A solvent-dependent stereoselectivity found for intramolecular [4 + 4] photocycloaddition of tethered 2-pyridones is concentration dependent, indicating that a dimeric structure with four hydrogen bonds plays a critical role in the observed cis selectivity found for nonpolar solvents. PMID- 10836038 TI - Solid-phase oligosaccharide synthesis: preparation of complex structures using a novel linker and different glycosylating agents. AB - [formula: see text] A beta-(1-->4)-linked trisaccharide was prepared in 53% yield on a polymer support using glycosyl phosphates and released by cross-metathesis of a novel linker to reveal the anomeric n-pentenyl glycoside. Heptasaccharide 33 was prepared in 9% yield in 14 steps. PMID- 10836039 TI - Consequences of acid catalysis in concurrent ring opening and halogenation of spiroketals. AB - [formula: see text] Lewis and/or Bronsted acid additives permit ring opening and halogenation of spiroketals at substantially reduced temperatures to produce omega-iodo enol ethers in improved yield and purity, which can undergo further reaction in the presence of distal electrophilic centers to give new steroid skeletons. PMID- 10836040 TI - Novel cyclic tripeptides and substituted aromatic amino acids via ruthenium activated S(N)Ar reactions. AB - [formula: see text] Chlorophenylalanines eta 6-complexed to ruthenium undergo SNAr reactions with a variety of nucleophiles to form substituted phenylalanines exemplified by 4b. Extension of these reactions to intramolecular ruthenium activated SNAr cyclizations led to three novel cyclic tripeptide systems (exemplified by 17 and 20). PMID- 10836041 TI - Gram-scale synthesis of (+)-discodermolide. AB - [formula: see text] A triply convergent, highly efficient second-generation synthesis of the potent antimitotic agent (+)-discodermolide (1) has been achieved on a 1-g scale. PMID- 10836042 TI - Desymmetrization by ring-closing metathesis leading to 6,8 dioxabicyclo[3.2.1]octanes: a new route for the synthesis of (+)-exo- and endo brevicomin. AB - [formula: see text] The 6,8-dioxabicyclo[3.2.1]octane skeleton is a common structural subunit in natural products. A conceptionally new strategy affording these structures is described for the syntheses of (+)-exo-brevicomin and rac endo- and enantiomerically enriched (+)-endo-brevicomin, employing desymmetrization of trienes derived from diols with C2 and meso symmetry via ring closing metathesis. PMID- 10836043 TI - A highly enantioselective conjugate reduction of 3-arylinden-1-ones using bakers' yeast for the preparation of (S)-3-arylindan-1-ones. AB - [formula: see text] The bakers' yeast reduction of 3-(1,3-benzodioxol-5-yl)-6 propoxy-1H-inden-1-one 4 has been shown to give (S)-3-(1,3-benzodioxol-5-yl)-2,3 dihydro-6-propoxy-1H-indan-1-one 6 in 65% yield with high enantioselectivity (> 99.0% ee), a key intermediate for the synthesis of the endothelin receptor antagonist SB 217242. In addition, the substituted 3-arylinden-1-ones 10a-e gave equally high enantioselectivity for the 3-arylindan-1-one products 13a-e. Mechanistic studies of the reaction indicate the operative pathway to be an asymmetric conjugate reduction, wherein the hydride transfer from NAD(P)H occurs from the Re-face of the indenone substrate. PMID- 10836044 TI - Synthesis of the Bycroft-Gowland structure of micrococcin P1. AB - [formula: see text] We describe the chemical synthesis of the accepted structure of micrococcin P1, a member of the thiostrepton group of antibiotics, and we show that this architecture does not correspond to that of the natural product. Methods developed during the present study should greatly facilitate ongoing efforts centering on the determination of the actual structure of microccin P1, in addition to being applicable to the synthesis of more complex thiostrepton congeners. PMID- 10836045 TI - Soluble-polymer supported synthesis of a prostanoid library: identification of antiviral activity. AB - [formula: see text] The prostaglandins are potent natural products taking part in many biological processes. The "convergent generation of diversity" from a "toolbox" of prostanoid components, augmented with additional polymer-supported transformations, can enable construction of valuable libraries. A parallel-pool strategy was used to assemble a small library of prostanoids. The inhibition of a herpes-family virus demonstrated the potential for new drug discovery. PMID- 10836046 TI - A catalytic enantioselective total synthesis of (-)-wodeshiol. AB - [formula: see text] (-)-Wodeshiol of > 99% ee has been synthesized from the alpha,beta-enone shown using a number of noteworthy steps including a novel C-C coupling reaction. PMID- 10836047 TI - 7-Azabicycloheptane carboxylic acid: a proline replacement in a boroarginine thrombin inhibitor. AB - [formula: see text] The synthesis of thrombin inhibitor 3, which incorporates conformationally constrained 7-azabicycloheptane carboxylic acid (1) as a proline replacement, is described. The inhibition constant (Ki(thrombin) = 2.9 nM) indicates that 1 is a reasonable replacement of proline in the formation of a beta-turn tripeptide mimetic. PMID- 10836048 TI - Chemo- and stereoselective reduction of an alpha-cyanoketone by bakers' yeast at low temperature. AB - [formula: see text] The bakers' yeast mediated reduction of 3-oxo-3 phenylpropanenitrile (1) proceeds at 4 degrees C to give exclusively (S)-3 hydroxy-3-phenylpropanenitrile (3) in 59% yield. This is in contrast to the corresponding reaction at room temperature which yields a mixture of reduction and alkylation products. This work demonstrates the use of low temperature to improve yeast selectivity. PMID- 10836050 TI - Regioselective Rh-catalyzed allylic amination/ring-closing metathesis approach to monocyclic azacycles: diastereospecific construction of 2,5-disubstituted pyrrolines. AB - [formula: see text] Regioselective rhodium-catalyzed allylic amination followed by ring-closing metathesis, using the Grubbs' catalyst, provides an expeditious route to monosubstituted azacycles. The enantiomerically enriched allylamine 1 can also be resubjected to the reaction sequence with (R)- and (S)-2b to facilitate the diastereospecific construction of 2,5-disubstituted pyrrolines 3/4. PMID- 10836049 TI - Molecular architecture via coordination: self-assembly of pseudohexagonal A2(3)X2(3)-macrocycles. AB - [formula: see text] The interaction of two complementary ditopic building blocks, each incorporating 120 degrees angles between the active coordination sites, in methylene chloride at room temperature results in the spontaneous self-assembly of platinum-based assemblies of hexagonal shape. PMID- 10836051 TI - Enantiopure N-acyldihydropyridones as synthetic intermediates: asymmetric synthesis of (-)-slaframine. AB - [formula: see text] An asymmetric synthesis of (-)-slaframine and N acetylslaframine has been accomplished starting from an enantiopure dihydropyridone building block. The oxygen-carbon bond at C-1 was incorporated with complete stereoselectivity by using an efficient phenylselenocyclocarbamation reaction. PMID- 10836052 TI - Inhibition of phosphatase activity by positively-charged cyclodextrins. AB - [formula: see text] Aminocyclodextrins are known to bind phosphate esters such as phosphotyrosine and p-nitrophenyl phosphate. This paper describes the inhibition of phosphate ester hydrolysis, as catalyzed by lambda-protein phosphatase and acid phosphatase, that is caused by such binding interactions. ROESY studies provide structural information about the cyclodextrin-aryl phosphate complexes. In addition, these experiments are used to generate approximations of the rates of dissociation of the noncovalent complexes. PMID- 10836053 TI - Reversal of regioselection in the sharpless asymmetric aminohydroxylation of aryl ester substrates. AB - [formula: see text] The asymmetric synthesis of beta-hydroxy-alpha-amino acids is reported which relies on the use of alpha,beta-unsaturated aryl ester substrates and the dihydroquinyl alkaloid ligand system (DHQ)2-AQN to control the regio- and enantioselectivity of the asymmetric aminohydroxylation (AA) process. alpha,beta Unsaturated ester substrates of type 1 have a significant effect on the substrate ligand recognition event which results in a reversal of regioselectivity in the AA reaction. PMID- 10836054 TI - Effect of substituents in directing the formation of benzochlorins and isobacteriochlorins in porphyrin and chlorin systems. AB - [formula: see text] A first synthesis of free-base fluorinated benzochlorins by acid-catalyzed cyclization of 20-(2-trisiloxy trifluoromethylvinyl)octaethylporphyrin++ + is achieved. Under similar reaction conditions, the purpurin-18-N-hexylimide analogues produced the corresponding fluorinated and nonfluorinated ethylidene-substituted isobacteriochlorins and fluorinated chlorin, respectively. The structure of the porphyrin based fluorinated benzochlorin was also confirmed by X-ray analysis. PMID- 10836055 TI - Nucleophilic metal complexes as acylation catalysts: solvent-dependent "switch" mechanisms leading to the first catalyzed Staudinger reaction. AB - [formula: see text] Catalytic acylation using complex transition metal salts MCo(CO)4 is demonstrated. Surprisingly, a solvent-dependent mechanistic "switch" results in a Lewis acid-based acylation mechanism in nonpolar media and a nucleophilic mechanism in polar organic media. These observations lead to the first example of a catalyzed Staudinger reaction to form beta-lactams. PMID- 10836056 TI - Synthesis of ultra-short-acting neuromuscular blocker GW 0430: a remarkably stereo- and regioselective synthesis of mixed tetrahydroisoquinolinium chlorofumarates. AB - [formula: see text] The stereo- and regioselective synthesis of ultra-short acting nondepolarizing neuromuscular blocker GW 0430 (5a) is described. Key steps involved the enantioselective transfer hydrogenation of imine 8 employing Noyori's catalyst, the stereoselective crystallization and methanolysis of trans bataines 11 and 12, and the stereo- and regioselective trans elimination of hydrogen chloride from 14. The latter transformation allowed complete control of the position of the chloro substituent and stereochemistry at the double bond of the linker in 15. PMID- 10836057 TI - Multicomponent linchpin couplings of silyl dithianes: synthesis of the Schreiber C(16-28) trisacetonide subtarget for mycoticins A and B. AB - [formula: see text] An efficient synthesis of trisacetonide (+)-11, the Schreiber C(16-28) subtarget for mycoticins A and B, is described. The key synthetic transformation entails a one-flask, five-component linchpin coupling tactic. PMID- 10836058 TI - Facile synthesis of the tricyclic core of sarain A. 3-Oxidopyridinium betaine cycloaddition approach. AB - [formula: see text] A new approach to a suitably functionalized tricyclic core of sarains has been developed by means of Katritzky's cycloaddition using 3 oxidopyridinium betaines. A key step was the regioselective differentiation of the two nearly identical hydroxy groups derived from oxidative cleavage of the double bond in 8 to afford 14. A stereocontrolled construction of the tricyclic core 20 of sarains containing the requisite side chain at C-3' was achieved by an intramolecular conjugate addition. PMID- 10836059 TI - Synthesis of modified oligodeoxyribonucleotides on a solid-phase support via derivatization of a selectively revealed 2'-amino-2'-deoxyuridine. AB - [formula: see text] High yields of oligodeoxyribonucleotides modified at the C2' position of site specifically incorporated 2'-amino-2'-deoxyuridine are obtained by photolytically unmasking the nucleophile in an otherwise protected solid-phase support-bound biopolymer. PMID- 10836060 TI - Total synthesis of (+)-laurencin: an asymmetric alkylation-ring-closing metathesis approach to medium ring ethers. AB - [formula: see text] The enantioselective total synthesis of (+)-laurencin 1 is achieved in 18 steps from (S)-(+)-4-benzyl-3-benzyloxyacetyl-2-oxazolidinone. The key steps in this synthesis are an asymmetric glycolate alkylation leading to acyl oxazolidinone 2 and a subsequent ring-closing olefin metathesis to construct the oxocene core of 1. The approach to medium ring ethers utilized in this synthesis provides a general and efficient route to the cyclic core of other marine natural products. PMID- 10836061 TI - Theoretical studies of the Wilcox molecular torsion balance. Is the edge-to-face aromatic interaction important? AB - [formula: see text] Molecular mechanics calculations with several different force fields and the GB/SA solvation model were carried out for the molecular torsion balance developed by Wilcox et al. to investigate the intramolecular nonbonded interactions between two aromatic rings. The preference is found to arise from a balance between intramolecular van der Waals interactions and solvation effects, with the latter favoring the unfolded conformation. PMID- 10836062 TI - Aggregation and C-N rotation of the lithium salt of N,N dimethyldiphenylacetamide. AB - [formula: see text] Two methyl 1H NMR signals for the Li salt of N,N dimethyldiphenylacetamide are observed at low temperature and assigned to the monomer and dimer. From line shape analysis, the dimerization constant (K1,2) is 40 +/- 10 M-1 at 200 K (delta G degree = 1.5 kcal mol-1, delta H degree = 0.8 kcal mol-1, delta S degree = 12 eu) and the activation parameters are delta H++ = 5.5 kcal mol-1 and delta S++ = -18 eu. The C-N bond rotation is too fast to observe on the NMR time scale, indicating a rotation barrier of less than 10 kcal mol-1. PMID- 10836063 TI - Synthesis of L-4,4-difluoroglutamic acid via electrophilic difluorination of a lactam. AB - [formula: see text] An enantiomerically pure bicyclic lactam proved to be an excellent substrate for electrophilic difluorination using N fluorobenzenesulfonimide. The resulting difluorinated lactam can be easily converted into L-4,4-difluoroglutamic acid. To the best of our knowledge, this is the first example of a synthetically useful electrophilic difluorination of an unactivated lactam. PMID- 10836064 TI - Asymmetric solid-phase synthesis of (3'R,4'R)-di-O-cis-acyl 3-carboxyl khellactones. AB - [formula: see text] We describe a practical parallel synthesis of (3'R,4'R)-di-O cis-acyl 3-carboxyl khellactones on a solid phase in high yield. The highlights of this synthesis include a Knoevenagel condensation, asymmetric dihydroxylation, catalyzed acylation, and product cleavage from the solid support. PMID- 10836065 TI - Bromoacetophenone-based photonucleases: photoinduced cleavage of DNA by 4' bromoacetophenone-pyrrolecarboxamide conjugates. AB - [formula: see text] 4'-Bromoacetophenone derivatives which upon excitation can generate monophenyl radicals capable of hydrogen atom abstraction were investigated as photoinducible DNA cleaving agents. Pyrrolecarboxamide-conjugated 4'-bromoacetophenones were synthesized, and their DNA cleaving activities and sequence selectivities were determined. PMID- 10836066 TI - Efficient synthesis of a porphyrin-N-tripod conjugate with covalently linked proximal ligand: toward new-generation active-site models of cytochrome c oxidase. AB - [formula: see text] A new-generation cytochrome c oxidase active-site model compound (4) featuring both a trisimidazolyl moiety and a proximal base has been designed and efficiently synthesized. During this study, a facile method based on the chemistry of a 4-magnesioimidazole derivative to synthesize 4-imidazolyl containing tripodal ligands (7) has been developed. PMID- 10836067 TI - Improvements in cross coupling reactions of hypervalent siloxane derivatives. AB - [formula: see text] The scope of the palladium-catalyzed cross coupling reaction of aryl halides with phenyltrimethoxysilane has been expanded to include aryl bromides, heteroaryl bromides, and aryl chlorides. A more general Pd(0) catalyst/ligand system has been developed to activate bromides: palladium(II) acetate (Pd(OAc)2) is activated with triphenylphosphine (PPh3) or tri-o tolylphosphine (P(o-tol)3) (1:2 molar ratio of Pd:phosphine). Coupling of aryl chloride derivatives required addition of 2-(dicyclohexylphosphino)biphenyl (Buchwald's ligand) to Pd2dba3 (tris-(dibenzylideneacetone)dipalladium(0)) (1:1.5 molar ratio of Pd:phosphine). PMID- 10836068 TI - Convergent C-glycolipid synthesis via the Ramberg-Backlund reaction: active antiproliferative glycolipids. AB - [formula: see text] A novel methodology has been developed, employing the Ramberg Backlund rearrangement and ionic hydrogenation to synthesize C-glycosides with high stereoselectivity at the anomeric center. The C-glycolipid 14b exhibits antiproliferative properties similar to those of O-glycoside analogue 14a. PMID- 10836069 TI - Counterions of BINAP-Pt(II) and -Pd(II) complexes: novel catalysts for highly enantioselective Diels-Alder reaction. AB - [formula: see text] Platinum and palladium chiral bisphosphine complexes and their counterion effects in asymmetric Diels-Alder reactions have been investigated. The reaction of cyclopentadiene and various bidentate dienophiles in the presence of a catalytic amount of Pt(II)- or Pd(II)-BINAP complex proceeds with excellent endo/exo selectivity as well as endo enantioselectivity (up to 99% ee). PMID- 10836070 TI - Asymmetric total synthesis of batzelladine D. AB - [formula: see text] The first enantioselective total synthesis of a batzelladine alkaloid is described. The central reaction in the synthesis of (-)-batzelladine D (2) is a tethered Biginelli condensation of a guanidine aldehyde and an acetoacetic ester to generate a 7-substituted-1-iminohexahydropyrrolo-[1,2 c]pyrimidine intermediate having the anti stereochemistry of the methine hydrogens flanking the pyrrolidine nitrogen. PMID- 10836071 TI - In vivo and in vitro investigations into the biosynthetic relatedness of the pseudopterosins. AB - [formula: see text] Both in vivo and in vitro techniques have been developed to test putative intermediates in the biosynthetic pathway to the pseudopterosins, antiinflammatory compounds isolated from Pseudopterogorgia elisabethae. Furthermore, specific activity data we have obtained indicate that pseudopterosin A is a precursor to pseudopterosins B, C, and D. We conclude that in the biosynthesis xylose is attached to the diterpene skeleton to produce pseudopterosin A and is then aceylated to form pseudopterosins B-D. PMID- 10836072 TI - Toward creation of a universal NMR database for the stereochemical assignment of acyclic compounds: the case of two contiguous propionate units. AB - [formula: see text] Using triol 1 as a representative example of natural products containing two contiguous propionate units, 13C and 1H NMR databases for the stereochemical assignment of acyclic compounds have been created. Chemical shift increments due to the presence of additional functional groups as well as solvent effects are discussed. PMID- 10836073 TI - Toward creation of a universal NMR database for the stereochemical assignment of acyclic compounds: proof of concept. AB - [formula: see text] Using the C.5-C.10 portion of the oasomycin class of natural products, the reliability and usefulness of an NMR database for the stereochemical assignment of acyclic compounds has been demonstrated. The predicted relative stereochemistry based on the NMR database has unambiguously been established via synthesis. PMID- 10836074 TI - [Cytotoxicity tests of stomatologic materials on cell cultures]. AB - Authors compare the benefits and drawbacks of agar diffusion tests (with neutral red and with thiazolyl blue) and the filter diffusion test recommended as the standards for cytotoxicity testing of dental materials and their eluates. It seems that there are no differences between these diffusion tests after the treatment for 24 hours. Authors discuss the observed differences in the toxicity between a solid form of the material (ANA 2000 DUO, Evicrol Solar LC) and its eluate. PMID- 10836075 TI - [Kinetics of ceftazidime in prophylactic administration during cardiopulmonary bypass]. AB - Serum ceftazidime levels were followed in 21 patients in which routine coronary bypass surgery with cardiopulmonary bypass was performed. Each patient received one gram of ceftazidime intravenously with anesthesia induction. Antibiotic concentrations were estimated using the microbiologic assay diffusion plate method. The average operation time was 220 +/- 41 minutes (range 130-310). The start of cardiopulmonary bypass was 86 +/- 21 minutes and the full flow time was 104 +/- 21 minutes after starting of ceftazidime application. It can be stated that the decline of ceftazidime serum levels after starting of cardiopulmonary bypass was faster in comparison with standard serum curves of this antibiotic. The concentrations of ceftazidime at the end of some operations were under the supposed minimal inhibitory concentrations for some microorganisms possibly implicated. No infection was recorded. PMID- 10836076 TI - [Treatment of keloids and hypertrophic scars with cryotherapy]. AB - Keloid and hypertrophic scars are relatively frequent not wanted consequences of surgical treatment. 145 patients with unsatisfactory skin healing were treated in our department during last 5 years. When compared with corticotherapy and laser therapy kryotherapy is very effective method. There were outstanding results with combination of cryotherapy and corticotherapy. PMID- 10836077 TI - [Health status of persons occupationally exposed to chromium, nickel, manganese and polycyclic aromatic hydrocarbons]. AB - Occupational environment monitoring and biological-medical monitoring of persons professionally exposed to welding fumes have been performed. Chromium, manganese and polycyclic aromatic hydrocarbons in welding fumes represents an important health risk. Pollutant concentrations found in metal welding fumes represented only fractions of those acceptable ones. Polycyclic aromatic hydrocarbons have been reached the concentration found in a busy road crossing in Hradec Kralove (compared with these as in Czech Republic no maximum acceptable levels for PAHs having been declared). Family, personal and occupational history have been taken. Health state including total haematological count, biochemical and cytogenetical changes of 19 stainless steel welders were checked-up. The level of mercapturates in urine were examined as well. The data were statistically compared with those of non exposed (control group). No changes witnessing the above mentioned risk factors influence on the haematological, biochemical and cytogenetical findings were ever proved. In conclusion, our results did not confirm an increased professional risk in this group of welders. PMID- 10836078 TI - [Biological monitoring of occupational exposure in welders of stainless steel. Immunologic methods]. AB - Biological monitoring of occupational exposure to toxic compounds enables an early detection of adverse health effects. Apart from the standard biological monitoring methods of occupational exposure represented by metabolites and enzymes activities analysis also immunological detection methods seem to be promising. The changes of immunological system are demonstrable extraordinary sensitive markers of load of organism. The process of welding belongs between important sources of pollution of working environment with heavy metals and other harmful compounds including polycyclic aromatic hydrocarbons. According to IARC classification the welding fumes are included into group 2B and are supposed as possible human carcinogens. Numbers of different epidemiological studies have proved relationship between welding and serious health disorders namely malignancies. We recorded a number of changes in immunological parameters when compared with those in control group in our study. The results of analysis were statistically evaluated. The statistically significant reduction of number of phagocytosis capable cells was found in the group of welders when compared with the control group. The levels of neopterin, beta-2-microglobulin and IL-1 beta have indirectly monitored cell component of immunological response. In the case of these markers a statistically significant increases were found in the group of welders. We found statistically significant IgM decrease and statistically significant IgA increase in humoral responses with the same group when compared with control group. PMID- 10836079 TI - [Polycyclic aromatic hydrocarbons. I. Environmental contamination and environmental exposure]. AB - Polycyclic aromatic hydrocarbons (PAHs) represent danger ubiquitous environmental pollutants. A lot of them have toxic and carcinogenic potential. Presented work summarises most of available data describing properties, origin and occupational and non-occupational sources of PAHs. Contamination of environment is described separately for air, water, soil, sediments and food. Possibilities of occupational and non-occupational exposure of persons are discussed and populations with potentially high exposures to PAHs are defined. The work is concluded by digest of regulations and guidelines regarding environmental contamination of PAHs. PMID- 10836080 TI - [Relation between month of birth and the manifestation of atopic diseases in children and adolescents]. AB - A total of 354 children with atopic diseases (asthma, pollenosis, dermatitis) were examined. Less than half of them (46.6%) had atopy in their families. Children of atopic parents often had two atopic diseases. Hypersensitivity to single allergen occurred in 66% (234/354) of patients and to more than one allergen in 34% (120/354) of patients. It seems that the season of birth might be important for children sensitized to pollens (born on the eve of pollen season). For the children sensitized to Dermatophagoides pteronyssimus season of birth was not decisive. PMID- 10836081 TI - [Analysis of lymphocyte immunophenotypes in neonates]. AB - Immune system development is not completed at the end of gestation, so newborns are not fully immunocompetent. In order to evaluate the neonatal immune system status and investigate the reasons for increased neonatal susceptibility to infections, the major lymphocytes subpopulations were studied in newborns comparing the results to adult controls. The cord blood from 21 term-newborns and the peripheral blood from 23 adults were analyzed using fluorochrome labelled monoclonal antibodies and two-color flow cytometry. The percentage of T lymphocytes was lower in newborns (64.9 versus 72.8% in adults), as well as the percentage of NK cells (4.8 versus 15.5%). On the contrary, the proportions of unlabelled cells were increased in term cord blood. The percentage of cytotoxic T lymphocytes was significantly lower in term-newborns as compared to the adult controls (17.8 versus 30.3%), and so were the percentages of activated T lymphocytes (0.3 versus 3.7%) and cytotoxic non-MHC restricted T lymphocytes (0.2 versus 1.8%). The expression of CD45R0 marker on neonatal lymphocytes was very low (1%). These characteristics of newborn lymphocytes phenotype are the result of inexperienced and partly undeveloped immune system. PMID- 10836082 TI - [Results of treatment of hip joint prosthesis-related infections]. AB - The article present results of the operative treatment of deep infections in the hip joint prostheses. Ninetieth patients treated from 1980 through 1998 were analysed. Gram-positive organisms were found in 65% of the cases and gram negative in 35%. Staphylococcus aureus were isolated in 50% of the cases. The two stage reimplantation procedure is performed in four patients. Excellent results were reported in three patients, and poor results in one patient. The Girdlestone pseudarthrosis as a definitive procedure is performed in fifteen patients. Good results were reported in two patients, fair results in twelve, and poor results in one patient. The mean shortening of the limb was 4.5 cm. The infection were controlled in 93% of the cases. Better functional results are obtained by the two stage reimplantation than the Girdlestone pseudarthrosis, though is an acceptable method of controlling infection. PMID- 10836083 TI - [Comparison between the rubber Carlens tube and the polyvinylchloride Robertshaw tube for endobronchial intubation]. AB - The aims of the study were to compare prospectively two double-lumen endobronchial tubes (DLTs): red rubber Carlens and left polyvinylchloride Robertshaw DLT with regard to difficulties in intubation and complications during one-lung ventilation. Sixty-two adult patients were randomly intubated with Carlens tube (31 patients) or Robertshaw tube (31 patients). Correct placement of DLTs was assessed using clinical signs and in some patients in the Robertshaw group fiberoptic bronchoscope was used. In the group intubated with the Carlens tube there were significantly more difficulties in passing the tube through the larynx (4 out of 31 patients), compared with the Robertshaw group (not a single patient). There was no difference in the number of patients with difficulties in placing the tube in the left main bronchus; however, the nature of the problem was more complex with the Carlens tube. The frequency of all the complications as well as the occurrence of the most common complications--hypoxemia and increased peak airway pressure, was not different between the groups. In both groups hypoxemia was related to the right side of the operation. Fiberoptic bronchoscope was used in 18 out of 31 patients intubated with the Robertshaw tube. Its use was not correlated with decreased incidence of complications compared to intubation in which tube position was assessed by clinical signs only. PMID- 10836084 TI - [Delivery with the foot and umbilical cord alongside the head]. AB - Compound presentation with a foot and umbilical cord alongside the head at the pelvis outlet in a primipara is described. A live femal newborn, birth weight 2850 g, birth length 49 cm, Apgar score 5/8/9, with normal postnatal development, was born by vaginal delivery. PMID- 10836085 TI - [High frequency current in endoscopic surgery and a new bipolar hook for endoscopic surgery]. AB - High frequency current is used in surgery for cutting the tissue and stopping bleeding. Its usage is more frequent in endoscopic than open surgery. "Polarity" (monopolar and bipolar) marks the number of the electrical poles on the application place. Monopolar current has been used more often. With its usage thermal injuries are possible: coagulation outside of the laparoscopic view, direct coupling, capacitive coupling, sparking, skin combustion, activity on the heart rhythm. Because of these complications, the bipolar current is more and more in usage, which reduces the number of complications or avoids them completely. Many bipolar instruments are available today. Bipolar hook for endoscopic surgery is a new instrument which unites the good characteristics of the hook and bipolar current. PMID- 10836086 TI - [Radiotherapy of lung cancer]. AB - The paper is aimed at approaching radiation therapy methods to physicians of other specialties and pointing to the potential of radiation therapy in the management of lung cancer patients. With the reference to its incidence and mortality rates, lung cancer ranks among the most frequent human malignant tumors. Therapy procedures for lung cancer depend upon tumor histology type, stage of disease and patient general condition. The said parameters therefore determine the application of surgery, radiation therapy and/or chemotherapy. In general, treatment results are usually rather poor, primarily due to lung cancer being the most frequently detected only as locally advanced or metastatic disease. Alike surgery, radiotherapy is a local form of treatment aimed at achieving local tumor control. This curative or palliative form of treatment is either applied alone or in combination with other treatment modalities. Irradiation is usually delivered by high energy photon beams from a telecobalt device or linear accelerator. The success of radiation therapy complies with the irradiation dose managed to be applied to tumor or tumor bed, which depends on patients general condition and site, size and spread of tumor. Radiotherapy with curative intent is applied in stage I, II and III non-small cell lung cancer patients with surgery being primarily applied in those with stage I and II. The efficacy of surgical treatment is to be improved by a combined-modality treatment. In stage III patients, who are more frequent than others, radical radiotherapy alone or in combination with chemotherapy is applied. Results of clinical trials report patients of relatively good general condition benefiting from combined-modality therapy. Palliative radiotherapy is to be applied in patients with stage IV non-small cell lung cancer. On the other hand, in patients with small cell lung cancer chemotherapy is the primary modality treatment. When the disease is limited to the lungs, the aim of radiotherapy is to optimize local control of the primary tumor. PMID- 10836087 TI - [Percutaneous tracheostomy]. AB - In the last few years numerous reports from intensive care units confirm that a nonsurgical percutaneous tracheostomy has successfully replaced elective conventional (surgical) tracheostomy. The majority of authors point out the advantages over surgical technique such as: the speed of procedure, the possibility of doing it at bedside thus excluding transport and the need for operating theatre, less infections around stoma, minor cosmetic defects and finally, more economical price. The article describes three, according to pertinent literature most often utilized techniques, i.e. percutaneous dilatation tracheostomy with dilators, percutaneous dilatation tracheostomy with forceps, and translaryngeal tracheostomy. Main differences, advantages and defects of each method are presented. Contraindications and eventual complications are discussed, as well as the utility of supporting methods for safer placement of tracheostomy tubes, such as endoscopy and ultrasonography. PMID- 10836088 TI - Minority issues in prevention: introduction to the special issue. PMID- 10836089 TI - Money doesn't talk, it swears: how economic stress and resistance resources impact inner-city women's depressive mood. AB - We examined the differential impact of chronic versus acute economic stress on depressive mood among a sample of 1241 low-income, single, European and African American women. Based on Hobfoll's (1988, 1989) conservation of resources (COR) theory, we predicted that acute resource loss would be more distressing than chronic economic lack. That is, although chronically impoverished conditions are stressful, the attendant resource losses created will be more distressing. We further predicted that mastery and social support would be more beneficial in offsetting the negative consequences of acute resource loss than the negative consequences of chronic economic lack, because acute loss creates identifiable demands that resources may address. Hence, we hypothesized that mastery and social support would show stress buffering effects only for material loss, not chronic lack. The findings generally supported the hypotheses, but mastery buffered only European American women's resource loss and social support buffered only African American women's resource loss. The findings are discussed in light of implications for prevention within theoretical and cultural contexts. PMID- 10836090 TI - Family conflict and children's internalizing and externalizing behavior: protective factors. AB - The current investigation examined whether the positive association of family conflict to adolescent depression and conduct problems is attenuated by maternal, paternal, and peer attachment, and maternal and paternal monitoring, within a low income, multiethnic sample of 284 adolescents. Parental attachment and monitoring moderated the link from family conflict to conduct problems but not depression; the relationships among family conflict, the hypothesized protective factors, and conduct problems were further modified by adolescent gender but not ethnicity. In general, higher levels of the hypothesized protective factors attenuated the relationship between family conflict and conduct problems for girls but exacerbated this relationship for boys. These findings suggest that, in general, parental attachment and monitoring served as protective factors for girls while serving as additional risk factors for boys in conflictual families. PMID- 10836091 TI - The roles of ethnicity and school context in predicting children's victimization by peers. AB - This study examines the prevalence, stability, and contextual correlates of peer victimization in a sample of African-American, Hispanic, and non-Hispanic White urban elementary school-age children. A total of 1956 children (40% African American, 42% Hispanic, and 18% White) attending any 1 of 14 public elementary schools located in one large and one mid-sized Midwestern city participated in this study. Peer ratings of victimization were obtained at two points in time, separated by a 2-year period. Findings revealed that risk for being victimized by peers varied by ethnicity and by school context. Hispanic children had lower victimization scores than did either African-American or White children. These findings, however, were moderated by school context, such that attending ethnically integrated schools was associated with a significantly higher risk of victimization for White children and a slightly lower risk of victimization for African-American children and did not affect the risk of victimization for Hispanic children. In addition, African-American children were less likely than Hispanic and White children to be repeatedly victimized by peers over time. The importance of considering ethnicity and context in explaining peer victimization is discussed and suggestions for preventive interventions and future research are provided. PMID- 10836092 TI - Do coping styles differ across sociocultural groups? The role of measurement equivalence in making this judgment. AB - Cross-sociocultural group measurement equivalency is an important issue that generally has not been studied in the coping literature. Measurement equivalency of the COPE (Carver, Scheier, & Weintraub, 1989) was assessed across two sociocultural groups, a sample of 100 Anglo middle-class divorced mothers and a sample of 122 low-income Mexican American/Mexican immigrant mothers. A series of restrictive confirmatory factor analyses revealed that seven of the COPE's subscales may be measuring the same underlying construct across populations. However, scores derived from the subscales may not represent the same magnitude of the construct in these two groups. This study makes an important first step in furthering the understanding of coping strategies in low-income Mexican American/Mexican immigrant mothers. This study also illustrates the importance of testing for measurement equivalency before conducting comparative research in disparate populations. PMID- 10836093 TI - Predictors of condom use in Mexican migrant laborers. AB - The purpose of this study was to explore predictors of condom use with occasional sex partners and regular sex partners, as well as carrying condoms in a new high risk group for HIV infection, Mexican migrant laborers. This study extends previous findings by (1) exploring additional predictors not previously examined, (2) utilizing a large sample of male and female Mexican migrant laborers, (3) carefully controlling for the effects of various demographic and lifestyle variables related to condom use, and (4) assessing the interactive effects of gender on predictors of condom use. Snowball sampling was used to survey 501 adult Mexican migrant laborers. Results revealed that condom use with occasional sex partners was predicted by carrying condoms and condom self-efficacy and that women were more likely to use condoms with occasional partners when both men and women knew someone with HIV/AIDS. Condom use with regular sex partners was predicted by procondom social norms, less negative attitudes toward condoms, not knowing someone with HIV/AIDS, and condom self-efficacy. Carrying condoms was predicted by procondom social norms, less negative attitudes toward condoms, condom self-efficacy, worry about contracting HIV/AIDS, and women were more likely than men to carry condoms when both men and women were married. Understanding these findings, future research directions, and implications for condom promotion strategies with Mexican migrant laborers are discussed. PMID- 10836094 TI - Cytology, synaptology and immunocytochemistry of commissural neurons and their putative axonal terminals in the dorsal cochlear nucleus of the rat. AB - The first binaural integration within the auditory system responsible for sound localization depends upon commissural neurons that connect the two symmetrical cochlear nuclei. These cells in the deep polymorphic layer of the rat dorsal cochlear nucleus were identified with the electron microscope after injection of the retrograde tracer, Wheat Germ Agglutinin conjugated to Horseradish Peroxydase, into the contralateral cochlear nucleus. Commissural neurons are multipolar or bipolar with an oval to fusiform shape. Few commissural neurons, most inhibitory but also excitatory, connect most of the divisions of the rat cochlear nuclei. The most common type is a glycinergic, sometimes GABAergic, moderately large cell. Its ergastoplasm is organized into peripheral stacks of cisternae, and few axo-somatic synaptic boutons are present. Another type of commissural neuron is a medium-sized, spindle-shaped cell, glycine and GABA negative, with sparse ergastoplasm and synaptic coverage. A giant, rare type of commissural neuron is glycine-positive and GABA-negative, with short peripheral stacks of ergastoplasmic cisternae. It is covered with synaptic boutons, many of which contain round synaptic vesicles. Another rare type of commissural neuron is a moderately large cell, oval to fusiform in shape, immunonegative for both glycine and GABA, and contacted by many axo-somatic boutons. It contains large dense mitochondria and numerous dense core vesicles of peptidergic type. Some labelled boutons, mostly inhibitory and probably derived from commissural neurons, contact pyramidal, cartwheel, giant and tuberculo-ventral neurons. The prevalent inhibition of electrical activity in a cochlear nucleus observed after stimulation of the contralateral cochlear nucleus may be due to commissural inhibitory terminals which contact excitatory neurons such as pyramidal and giant cells. Other inhibitory commissural terminals which contact inhibitory neurons such as cartwheel and tuberculo-ventral neurons, may explain the stimulation of electrical activity in the DCN after contralateral stimulation. PMID- 10836095 TI - Lamellar inclusions and trilaminar substance in the parabronchial epithelium of the quail (Coturnix coturnix). AB - The fine structure of the epithelial cells of the parabronchus and their secretory products have been the subject of many studies and have given rise to considerable controversy about their configuration and ultrastructure. The aim of the present study was to investigate the mode of formation and discharge of lamellar bodies of granular cells and the trilaminar substance produced and discharged by the embryologically related squamous atrial and respiratory epithelial cells. The material for light and transmission electron microscopic analysis was collected from 10 mature quail and 3 individuals aged 2 days. The parabronchial atria harbour two ultrastructurally distinct types of epithelial cells. The granular cells (analogous to type II cells of the mammalian pulmonary alveolus) produce and discharge balls of lamellar bodies. The squamous atrial cells produce and discharge sheets of trilaminar substance sandwiched between long tentacle-like processes, viz. the microvilli. The infundibula and air capillaries of the gas exchange tissue are invested with squamous respiratory cells which extend very thin, long processes that cover the air capillaries and constitute, together with the blood capillaries, the blood-air barrier. The squamous respiratory cells produce and discharge trilaminar substance as an extracellularly located acellular lining layer which is found in close contact with their cell membrane. Both squamous atrial and respiratory cells hence synthetize and discharge trilaminar substance, the basic unit of which has the ultrastructural appearance of a 7.5 to 8.0 nm membrane unit. The formation of trilaminar substance originates in the agranular endoplasmic reticulum, while the origin of the lamellar bodies of granular cells is related to the granular endoplasmic reticulum, the Golgi complex and the multivesicular bodies. Their structural unit is composed of a 4.5 to 5.0 nm thread-like structure which is concentrically arranged around a spherical core of granular substance. PMID- 10836096 TI - Morphological characterization of the seminiferous cycle in the goat (Capra hircus): a histological and ultrastructural study. AB - The cycle of spermatogenesis/seminiferous cycle was investigated in the goat testis using both light and electron microscopy techniques. Using the various cell associations and the accompanying changes in spermatid shape and location, the cycle was divided into eight (8) successive stages. The cycle began with the accomplishment of spermiation (stage 1) and ended with apical migration and close attachment of late maturation phase spermatids at the Sertoli cell apex accompanied by adluminal retention of residual bodies with dense staining inclusions (stage 8). The early stages of the cycle (stages 1-4) were therefore characterized by the presence of only one generation of spermatids, the second one appearing only after the division of secondary spermatocytes in stage 4. Consequently, stages 5-8 had two generations of spermatids; Golgi or cap phase as well as maturation phase spermatids. Although stages 5 to 7 appeared as distinct entities, stages 6 and 7 were rather short-lived and considered as continuations of stage 5. Therefore, the 8 stages of the cycle in the goat were further condensed into 6 main divisions. The duration of each stage was estimated by the frequency of occurrence in sections. Among these, stage 1 had the highest frequency (34%) followed by stages 5-7 (27%). Stages 8 and 4 had the shortest frequency (up to 9%) while stages 2 and 3 had 13% and 12% respectively. These results indicate that, like most domestic species, goats have a cycle of 8 stages with 6 main divisions, the longest being stage 1. PMID- 10836097 TI - Acetylcholinesterase activity in rat thymus after immunostimulation with interleukin beta. AB - The occurrence and distribution of Acetylcholinesterase (AChE) activity were examined in the thymus of normal and immuno-stimulated adult and aged rats using biochemical and enzymehistochemical methods. Specific AChE reactivity was found primarily in the arteries and, to a lesser extent, in the veins. Only a small amount of activity could be observed in association with the subcapsular and medullary part of the parenchyma and nerve fibers. Our findings indicate that AChE activity in the rat thymus increases after treatment with interleukin beta. In fact treatment with interleukin beta induces an increase of protein content, of the amounts of AChE biochemically assayed and at the levels of AChE histoenzymatically stained. Furthermore, staining of the different structures of the thymus in treated or untreated rats shows that the significant modifications concern the parenchyma, the structures resembling nerve fibers and the whole thymus, while only small changes are observed in AChE activity located in the walls of arteries, veins and lymphatic vessels. PMID- 10836098 TI - Production of epidermal growth factor in human prostatic cells cultured in vitro. AB - The Epidermal Growth Factor (EGF) plays an important role in the regulation of in vitro growth of prostate cells inducing a strong mitogenic effect. Nevertheless in our previous study we observed that the treatment of human hypertrophic prostate cell line U285 with exogenous EGF produces a restricted effect on the cellular growth rate. This phenomenon could be due to the capacity of the cells to produce EGF. In this study we aimed to verify this hypothesis by evaluating the presence of mRNA of EGF and EGF receptor (EGF-R) and of their translation products in U285 cells, before and after the treatment with suramin and exogenous EGF. Moreover we studied the effects exerted by these substances on the proliferative rate of the cells U285 after different treatment protocols. The presence in the cells of mRNA for EGF and EGF-R and of their translation products was demonstrated by means of reverse transcription polymerase chain reaction (RT PCR) and immunocytochemical methods respectively. The modification of growth rate induced by these drugs was studied by FRAME Cytotoxicity Test. The operative modalities adopted to carry out these growth assays tended to 1) focus the effects of suramin in relation to in vitro cellular growth phase; 2) verify the reversibility of its effects; 3) ascertain if it was possible to antagonize the action of suramin by adding exogenous EGF. The results obtained from the RT-PCR showed the presence, in the control cells and in the treated ones, of mRNA coding for EGF and EGF-R. The immunocytochemical analysis indicated that 20% of the control cells are EGF positive, and 83% are EGF-R positive, confirming the results obtained with RT-PCR. Moreover, these stainings showed that the treatment with EGF does not significantly modify the percentage of cells marked by the anti EGF antibody, while treatments with suramin and suramin plus EGF double this percentage. None of the treatments modifies the percentage of EGF-R positive cells. The growth assays showed that the exposition to highest doses of suramin in the first 24 h of cultures causes a decrease (p < 0.05) of the cellular proliferation during the following 48 h and 72 h and that these effects are irreversible. Moreover, a contemporaneous exposition of the cells to EGF and suramin at seeding strengthens the cytotoxic action of the last drug. To sum up, the demonstration of the presence in the U285 cells of mRNA coding for EGF and EGF-R and of the corresponding proteins, confirms the hypothesis that these cells can produce EGF. Moreover, the cytotoxicity experiments allowed a focusing of the role of the endogenous EGF in the regulation of the U285 cells proliferation and confirmed the importance of biological events that take place in U285 cells during the first 24 h of culture. PMID- 10836099 TI - Morphological variability and specializations in bovine extraocular muscle spindles. AB - Bovine extraocular muscles were examined to determine whether the structure of their muscle spindles was notably different from those commonly encountered in mammalian limb muscles. Extraocular muscle spindles on the whole were shorter, and intrafusal fiber counts/spindle were more variable than in somatic muscles. No pronounced nuclear bags were seen in intrafusal fibers. Based on cross sectional areas, intrafusal fibers in extraocular muscles could be loosely categorized as small or large types. Small fibers expressed more neonatal/fast myosin heavy chain and less embryonic myosin heavy chain than large fibers. When incubated for myosin ATPase, about 70% of the large fibers and 15% of the small fibers in spindles presented profiles that were characteristic of type I extrafusal fibers, and not of nuclear bag or nuclear chain fibers. The ratio of number of small intrafusal fibers to number of large intrafusal fibers in extraocular spindles was on average greater than the ratio of nuclear chain fibers to nuclear bag fibers that is typical for limb spindles of rodents and cats. Structural modifications at muscle spindle sensory regions, extrafusal-like fibers and intrafusal-like fibers with few equatorial nuclei and many myofibrils, may produce distinct afferent signals that are appropriate for sensorimotor integration in the specialized extraocular muscles. PMID- 10836100 TI - Morphological changes in mouse accessory sex glands following neonatal estrogen treatment. AB - A single injection of beta-estradiol 17-cypionate into the mice within 5 hr after birth induced inflammation in all prostate lobes and the seminal vesicles. Neutrophils emigrated into the lumen through the basal lamina and epithelium of the seminal vesicle and the anterior prostate. Local infiltration of lymphocytes was observed in the stroma and epithelium of ventral prostates. Lymphocytes penetrated through smooth muscle cells into epithelium. This could support the hypothesis that smooth muscle cells are the target of the estrogen action of prostates in estrogenized animals. PMID- 10836101 TI - Comparative angioarchitectural formation seen in microvascular cast specimens of the fungiform papillae in the weaning period and adult rat tongue. AB - The purpose of this study was an attempt to identify the functional and comparative angioarchitectural differences between the sexes, and the developmental processes seen in microvascular cast specimens (MVCS) of the formation of the fungiform papillae (FuP) geometrically and regularly distributed on the anterodorsal surface in the weaning period and in the adult rat tongue. The basic microvascular structure seen in the MVCS of FuP of both the weaning and adult rat tongue consisted of several ascending (As. b) and descending branches (Ds. b) and a loop structure (L. st), and the cylindrical network structure of the L. st made up of the open-hole formation of the C form of the upper and lower microvascular structures. In the lateral view, the MVCS of FuP has a bamboo basket-like shape, and by means of the three-dimensional expansion of the surface area, effectively plays an assistant functional role in receiving the taste of foods and liquids. There were obviously no sex and morphological differences in the developmental process as to shape, but there was some difference between the weaning period and the adult rat in size in the MVCS of FuP. PMID- 10836102 TI - Association of the truncus bicaroticus, common trunk of the left sub-sclavian and vertebral arteries, and retro-esophageal right subclavian artery. Case report. AB - We present a rare case of combined anomalies of the aortic arch. At its convexity, the arch of aorta gave rise, from right to left, first to the truncus bicaroticus and then to the common trunk of the left subclavian and left vertebral artery. A retro-esophageal right subclavian artery arose from the dorsal aspect of the arch, distally from the common trunk of the left vertebral artery and left subclavian artery. The combination of these anomalies has not been described before and may have important clinical implications. PMID- 10836103 TI - Harmonic analysis of footprint symmetry in healthy adolescents. AB - In the present study, the within-subject normal symmetry of footprint shape and size was analyzed from a mathematical standpoint. On the standardized left and right footprints of 83 adolescents (46 boys, 37 girls) aged 12-15 years (mean 13 years), the outline of each foot excluding the toes was identified and automatically digitized by a computerized video analyzer. Only those subjects with both left and right continuous footprints were further analyzed (36 boys, 26 girls). The footprint area was computed. The footprint shape, independent of its size, was quantified using the elliptic Fourier analysis with a 20-harmonic truncation. The symmetry in shape was quantified on an intra-subject basis by calculating a morphological distance D between the mathematical reconstructions of the left and right footprints of each subject. Symmetry in size was assessed by right-to-left area ratio. Subjects were grouped for sex, and the mean values computed. Mean footprint area was significantly larger in boys than in girls (p < 0.05). Asymmetry in size (area ratio) was 1.01 in girls, 1 in boys. Within subject symmetry in footprint shape appeared high, with mean morphological distances of 5.95 in girls, and 6.06 in boys. No consistent associations between footprint symmetry and age, height and body weight, or shoe size were found. The mean size-independent shapes of the male and female left and right footprints were also calculated. Together with the analysis of individual asymmetry, they could be used as quantitative parameter in clinical diagnosis. PMID- 10836104 TI - Prenatal growth of the swine auricle. AB - The aim of this article was the morphological, topographical and metric illustration of the growth of the swine auricle during the prenatal period. The experiment was conducted on 188 pig fetuses of known ages (42nd-115th day of gestation), divided into seven age groups. On the 42nd day of gestation the auricle was seen to be a triangular ear-flap covering the orifice of the external auditory meatus. Among the 43-49 day-old fetuses the rostro-ventral growth of the ear-flap was noted as well as its extension. The rostro-ventral "descensus" of the orifice of the external auditory meatus appeared in the material studied. The growth of the bilateral auricles was symmetrical during the whole period investigated. The growth in length of the whole fetus seemed to be relatively less marked than the increase in the auricular height and width. The increase in the auricular height was greater than the growth in width of the auricle. The measurements of the auricle were strongly correlated with the total length of the fetus during the whole period investigated. PMID- 10836105 TI - [Management of severely head injured patients during the first 24 hours]. PMID- 10836106 TI - [Fiberoptic bronchoscopy during noninvasive positive-pressure ventilation in patients with chronic obstructive lung disease with hypoxemia and hypercapnia]. AB - OBJECTIVES: To assess the feasibility and safety of non invasive positive pressure ventilation (NIPPV) via a face mask to performing fiberoptic bronchoscopy (FOB) in patients with COPD contraindicating FOB in spontaneous ventilation. STUDY DESIGN: Clinical, prospective, open, non comparative trial of feasibility. PATIENTS: Ten consecutive COPD patients (71 +/- 5 year-old, PaO2 = 53 +/- 13 mmHg and PaCO2 = 67 +/- 11 mmHg), without any sign of acute respiratory failure, admitted to the intensive care unit for pneumonia requiring a bronchoalveolar lavage (BAL). Including were: PaO2 < 70 mmHg despite nasal O2 delivered at 3 L.min-1, PaCO2 > 50 mmHg, improvement of SpO2 with NIPPV before FOB. METHODS: Topical anaesthesia of the nose and pharynx was obtained with a 5% lidocaine spray. NIPPV was administered using a ventilatory support system Evita 4 (Drager) applied through a full facial mask secured to the patient with elastic straps. Patients were first allowed to acclimate for 5 min with NIPPV (IPAP = 16 cmH2O, EPAP = 0 and trigger of 0.3 cm H2O while a FIO2 kept at 0.7). A T-adapter Medisize (Peters) was attached to the facial mask. The tip of the FOB was inserted through the nose. Topical anaesthesia of the vocal cords was obtained with 1% lidocaine solution (3 mL). The FOB was inserted into the trachea up to a bronchial sub-segment. BAL was performed by instillation of 100 mL of saline solution. After FOB, the NIPPV was maintained for 5 min. Heart rate, SpO2 were measured continuously and arterial pressure at 2 min intervals. Arterial blood gas values were obtained just prior NIPPV and after 15 min and 60 min NIPPV disconnection. RESULTS: FOB duration was 11 +/- 4 min. SpO2 significantly improved during FOB (from de 91 +/- 4.7% to 97% +/- 1.7) without decrease of oxygen saturation lower than 90%. There were no changes in PaCO2 and PaO2 during the hour following the end of procedure. FOB under NIPPV was performed in all patients without complications and was very well tolerated in eight patients. After NPPV disconnecting, one patient required again NIPPV for 15 min. No patient required endotracheal intubation within 24 hours. All patients survived. CONCLUSION: Application of NIPPV during FOB is a safe technic for maintaining adequate gas exchange in hypoxaemic and hypercapnic COPD patients not in acute respiratory failure. After the end of the procedure a close surveillance in the intensive care unit is essential. PMID- 10836107 TI - [Evaluation of transfusion strategies during craniosynostosis repairs]. AB - OBJECTIVE: To evaluate the changes in transfusion practices during craniosynostosis surgery in children, with consideration of the transfusion associated risks. STUDY DESIGN: Retrospective study. PATIENTS: The study included 64 consecutive craniosynostosis repairs by the same neurosurgical and anaesthetic team, over a period of 17 years. METHODS: The children were allocated into two groups. In group I (1980-1991), blood loss was compensated precisely from scalp incision on. In group II (1992-1996), transfusion was only started when blood loss crossed a calculated limit considered as acceptable. The mean criterion of judgment was the quantity of transfused blood in comparison with the theoretical blood volume. Mean preoperative and postoperative haemoglobin concentrations, estimated blood losses, volume of replaced blood and number of transfused patients were compared between the two groups with a two tailed Student's t test. RESULTS: The rate of non transfused children increased from 6% before 1991 to 39% after 1992%; conservely the rate of postoperative transfusions increased from 3 to 39%. CONCLUSION: The risks of blood transfusion favoured the development of new of more restrictive transfusion practices, even in haemorrhagic surgery. To limit blood transfusion in craniosynostosis surgery, a harmonious cooperation between surgeons and anaesthetists is essential and blood replacement must be based on a definite determination of the acceptable blood losses. PMID- 10836108 TI - [Impact of an information booklet on patient satisfaction in anesthesia]. AB - OBJECTIVE: The aim of this study was to evaluate the satisfaction of adult surgical patients before and after an information booklet concerning anaesthetic techniques and complications, and postoperative management has been proposed during preanaesthetic visits. STUDY DESIGN: Survey. METHOD: Three surveys have been performed using an anonymous questionnaire of 25 items with graduated answers (very good, good, bad, very bad, no opinion) concerning patient satisfaction on structure, physician behavior, information and well being. As the results of the first two surveys were not significantly different, an information booklet was proposed to patients during preanaesthetic visits, then a third survey ("After") was realized. The results of this survey were compared to the combined results of the first two surveys ("Before"). RESULTS: In the "before" survey, 60 to 76% of the patients were satisfied (very good and good) of the information delivered during preanaesthetic visits. The proportion of patients satisfied of the information concerning anaesthetic techniques, transfusion and recovery were significantly higher in the "after" survey (P < 0.05). CONCLUSION: An information booklet on anaesthesia increases patient satisfaction on information provided during preanaesthetic visits. PMID- 10836109 TI - [Cerebral arterial air embolism associated with pneumothorax in a patient with pressure support ventilation]. AB - A 20-year-old woman was admitted to the ICU following a road traffic accident. She had a periorbital haematoma with a normal cerebral state on CT-scan, intrahepatic and intrasplenic haematomas and several fractures of the limb that were fixed on day 3. Twenty-four hours later, she developed a cough with symptoms of decerebration The head CT-scan showed diffuse pneumocephalus suitable with cerebral air embolism and the chest X ray a right tension pneumothorax. A chest tube was inserted. Despite the lack of hyperbaric oxygen therapy the patient recovered fully. The pneumocephalus disappeared totally after 48 hours. Seven days later, CT-scan of the head with coronal slides showed an orbital floor fracture associated with an intra-orbital aerocele. PMID- 10836110 TI - [Severe pulmonary embolism with two indications for inferior vena cava filter placement]. AB - A case of severe pulmonary embolism, treated with thrombolysis, and complicated by a haemorrhagic shock (peritoneal bleeding after a spleen trauma) is reported. A paradoxical renal artery embolism occurred, due to a patent foramen ovale. The benefit of a inferior vena cava filter insertion in case of paradoxical embolism is discussed. PMID- 10836111 TI - [Failure to recover after anesthesia attributed to a transient dissociative state]. AB - Failure of a patient to recover rapidly after anaesthesia is a rare event, due to various causes, including medication related effects, metabolic disturbances or neurologic insults. We report a case in a middle aged physically healthy woman who failed to recover promptly after intravenous anaesthesia. Propofol was administered for 20 minutes. Results of neurologic assessment were normal, as were laboratory tests and CT-scan. Four hours after completion of the anaesthesia, return to consciousness occurred spontaneously. The patient had a history of depressive symptoms. However the psychiatric assessment was unremarkable. We hypothesise that she developed a transient dissociative disorder. PMID- 10836112 TI - [Epidemiology of severe cranial injuries]. AB - Head injuries (HI) are one of the most common causes of death, morbidity and disabilities in young adults. Epidemiological studies allow a quantitative estimation in terms of incidence and a qualitative estimate for the identification of risk factors in specific populations. These estimates may enable appropriate prevention programs. Estimates of annual incidence rates depend on territories, periods and methodological tools. Annual rates for hospitalized patients are found between 150 and 300/100,000 inhabitants. Severity of HI can be assessed by the Glasgow Coma Scale (GCS), the Abbreviated Injury Scale (AIS) or the Post-Traumatic Amnesia duration. Annual incidences of severe HI will depend on the selected score: around 25/100,000 inhabitants for cerebral trauma with intracranial injuries, around 9/100,000 for the most severe HI, with an AIS maximum of 5 with coma. The male:female ratio increases with degree of severity. Traffic accidents were the most frequent cause of HI. Many patients have associated injuries, worsening the outcome. Some risk factors are considered. Preventive measures are mainly conducted for traffic accidents, and include speed limit and regulations on helmet or seat belt use. Results of these measures are analysed. PMID- 10836113 TI - [From the macroscopic lesion to cellular ischemia]. AB - Traumatic brain injury (TBI) constitutes a major health and economic problem for developed countries, being one of the main causes of mortality and morbidity in children and young adults. Because of the immense importance and future consequences of TBI, the physician who sees a patient soon after brain injury must have a complete understanding of the pathophysiology and develop a practical knowledge of initial management of such patients. TBI may have intracranial and systemic effects that combine to give overall cerebral ischaemia. Injury to the nervous system is characterised by a stereotypic pattern, irrespective of the primary injury. The primary injury initiates a multitude of inflammatory cascades resulting in secondary brain injury, the effect of which is as important as the primary injury. This period of brain inflammation can last up to three weeks and renders the brain more susceptible to the effects of systemic insults such as hypotension, hypoxia and/or pyrexia. It has been shown in postmortem examination of patients dying from severe TBI that more than 90% had evidence of secondary ischaemic damage. The concept of 'cerebral protection' has been extended to encompass pretreatment of secondary injury. Preventing and treating cerebral ischaemia is the main goal of initial management of head-injured patients. Initial care focuses on achieving oxygenation, airway control and treatment of arterial hypotension. PMID- 10836114 TI - [Prehospital management of patients with severe head injuries]. AB - Advanced prehospital emergency medical care of patients with a severe head injury must essentially focus on the impact of secondary cerebral insults of systemic origin on the outcome. The first objective of prehospital care is to prevent hypoxaemia and hypercapnia. Therefore, all patients with a Glasgow Coma Scale score equal to or lower than 8 must be treated with endotracheal intubation and controlled ventilation under continuous monitoring of SpO2 and PETCO2. Treatment is similar in head-injured patients with significant deterioration of consciousness level, seizures, respiratory distress, or severe facial and thoracoabdominal injuries. The endotracheal tube is inserted by the orotracheal route under direct laryngoscopy, after a rapid induction sequence of anaesthesia and immobilization of the cervical spine in neutral position. For the induction of anaesthesia in these high-risk patients (full stomach, unknown medical history, deteriorated haemodynamic status), etomidate and suxamethonium are the preferred agents. Sedation is maintained with an hypnoticopioid association (fentanyl). Simultaneously, the main goal is the maintenance of an optimal cerebral perfusion pressure, as arterial hypotension severely worsens cerebral ischaemia. Volume loading is accomplished with 0.9% saline and hydroxyethyl starch. PMID- 10836115 TI - [Role of the regulation in the management of patients with severe head injuries]. AB - The regulator of the emergency medical ambulance service is involved in the various steps of the initial management of severe head trauma patients: handling calls, basic life support, prehospital advance life support, transport and hospital admission. The management is rapid (helicopter transports) coherent and adapted (adherence to the guidelines for severe head injury), and considers of local difficulties (geographical, possibility of admission to trauma centres), with the aim of improving the outcome of severely head-injured patients. PMID- 10836116 TI - [Management of severe head-injured patients in the first 24 hours. Resuscitation and initial diagnostic strategy]. AB - Limitation of secondary insults after severe head injury is a permanent concern during the early phase of head trauma management. The objectives are to maintain mean arterial pressure between 80 and 100 mmHg, to avoid hypoxaemia, and to maintain arterial PCO2 near to 35 mmHg. Volume loading can be necessary to improve arterial pressure, and is carried out with isotonic critalloid (NaCl 9/1000) or colloids, with the exclusion of all hypotonic solutions (Ringer lactate or glucose). The use of catecholamines is reserved for patients with unstable haemodynamics despite an adequate volume loading. The rapid sequence induction is recommended for endotracheal intubation and is followed by continuous analgesia-sedation to keep patient-ventilator dysynchrony, but without compromising haemodynamic objectives. Mannitol is used in case of life threatening intracranial hypertension. Conversely, specific treatment of intracranial hypertension, especially hypocapnia, is not recommended. Initial diagnostic procedures include cerebral tomodensitometry (TDM). However, TDM may be delayed in case of haemorrhage, which requires a rapid treatment. Intrahospital transport for additional explorations risks secondary insults, and thus requires close monitoring to detect and treat in due time all adverse events. This monitoring includes invasive arterial blood pressure assessment, use of continuous capnography and repeated arterial blood gas measurements. The usefulness of transcranial Doppler for initial management of head-trauma patients needs further evaluation. PMID- 10836117 TI - [Head and brain injuries. Place of imaging]. AB - This article considers the various mechanisms of brain injury and specifies the most efficient radiologic technique for assessing patients, depending on clinical presentation. The brain injuries include either extracerebral and intracerebral lesions. The former require rapid diagnosis and therapy and the latter determine management in an intensive therapy, unit and outcome. Standard X-rays are obsolete. The CT, rapidly performed, is the most relevant imaging procedure for surgical lesions. Cortical contusions and diffuse axonal injuries are underestimated by CT and best depicted by MRI. Only late MRI has a strong correlation with neuropsychological outcome. In terms of prognosis, MRI needs to be evaluated. The indications include: a) unstable neurological status: CT; b) moderate head injury: CT may help to decide hospital admission; c) severe head injury: initial CT may be followed by MRI; d) long-term consequences: MRI. Special Indications: a) angio-MRI: suspicion of vascular lesion; b) CT with thin slices and bone window: depressed skull fracture; c) teleradiology (image transfer): to decide a patient transport from a peripheral hospital to a neurosurgical centre. In conclusion, CT remains the first-line examination to detect immediately life-threatening lesions. MRI is the examination of choice for full assessment of brain lesions. PMID- 10836118 TI - [Management of severe head injuries during the first 24 hours, in the emergency department, in neurosurgery]. AB - In France, the role of the neurosurgeon in the emergency department depends on local health care policies and geographical constraints. Some departments include a neurosurgical team with a dedicated operating room. In others, a neurosurgeon can be reached by phone, possibly with an image transfer. Whatever the case, it should be possible to remove on site and without delay an intracranial haematoma, which is most often an extradural injury. The management of a haematoma of the posterior fossa or a bleeding dural venous sinus would be difficult for a surgeon not qualified in neurosurgery. The optimal situation is the presence of a neurosurgeon in the medical team admitting patients with a severe head or spine trauma, for assessment of the neurological status, or interpretation of radiological explorations, insertion of an intracranial pressure monitoring device. Besides the extradural haematoma, other injuries such as an acute subdural haematoma, a haematoma associated with a contusion, an acute hydrocephalus, a depressed fracture of the skull, or a craniocerebral wound, also require an emergency decompressive procedure. PMID- 10836119 TI - [Analysis of indications fo interhospital transfer]. AB - In France there are no clear guidelines for the transfer of severely head-injured patients, due to the difficulties of combining the requirements of a safe transfer with the necessity of a rapid, adapted and specialized management. These various aspects are discussed in order to facilitate the definition of local strategies for the initial orientation of severely head-injured patients to a centre adapted for severe head trauma management. PMID- 10836120 TI - [Monitoring of severely head-injured patients during the first 24 hours]. AB - Monitoring of severely head-injured patients is essential to optimize cerebral haemodynamics and thus to limit intracranial hypertension and to prevent the occurrence of secondary systemic cerebral injuries. It includes continuous measurement of intracranial pressure, mean arterial pressure and venous jugular oxygen saturation. Assessment of circulatory velocity in the arteries of the polygon of Willis allows identification of intracranial hypertension, and later, a vasospasm in case of subarachnoid haemorrhage. Near infrared spectroscopy is an indirect indicator of cerebral oxygenation. This technique has not yet been validated. Direct tissue measurement of cerebral oxygen content is the most recently developed monitoring tool. Its clinical range of application has still to be specified. These basic monitoring techniques are a standard. They are essential for an undelayed and efficient treatment of complications occurring in head trauma patients. PMID- 10836121 TI - [Vascular loading in the first 24 hours following severe head injuries]. AB - The main goal at the acute phase of head injury is to prevent a decrease in blood pressure, which promotes cerebral ischemia. Volume loading is therefore frequently indicated. A normal or increased plasma osmolarity should be maintained. Thus hypotonic fluids should be avoided. Hyperglycaemia is also a risk factor for brain injury and glucose use has to be restricted in the first hours after trauma. Isotonic saline 0.9% is the first solution to be infused. Lactated Ringer solutions are mildly hypotonic as approximately 114 mL of free water is contained in each litre of the solution. Isotonic colloids are indicated to replace blood losses, but have no advantage over cristalloids, concerning the development of cerebral oedema. Fluid restriction minimally affects cerebral edema. Because of the severe consequences of hypovolaemia and hypotension, fluids should not be restricted until haemodynamic stability is achieved. Hypertonic fluids rapidly restore intravascular volume and decrease intracranial pressure. Although they probably have a place in prehospital intensive therapy, the demonstration of their benefit is still lacking. Monitoring of intravascular volume is essential. Continuous arterial pressure and central venous pressure monitoring are mandatory. New monitoring techniques as the measurement of systolic pressure variations or transoesophageal Doppler echocardiography will probably find a place in the management of trauma patients in the near future. PMID- 10836122 TI - [Management of severely head-injured patients during the first 24 hours. Which specific therapeutics?]. AB - Intracranial and systemic mechanisms of the secondary brain lesion are the targets of specific therapy for the head-injured patient. Recommendations for good clinical practice have recently defined the role of the main therapeutic measures. There is no indication for corticosteroids in head injury. Mannitol is the first-choice therapy for increased intracranial pressure, and barbiturates are still considered as a rescue therapy in case of refractory intracranial hypertension. The place of hypothermia remains to be defined. Although controversial, optimized hyperventilation, induced systemic hypertension and vasoconstrictive therapy are optimally used under multimodal monitoring. New therapeutic perspectives, aimed at controlling biochemical disorders at a cellular level, are under investigation, but are still inconclusive at the present time. PMID- 10836123 TI - [Ventriculoperitoneal shunt and pleural effusion]. PMID- 10836124 TI - [Tracheal wound after intubation with rigid mandrin during reduction of a dislocated shoulder under general anesthesia]. PMID- 10836125 TI - Targets of drug action. AB - This article considers early work from the author's laboratory on muscarinic receptor specificity, subtypes, and conformational variability, with the use of nuclear magnetic resonance in pharmacology and the conformational variants of dihydrofolate reductase and general questions of receptors. It also considers some current approaches to drug development and receptor function, particularly as influenced by increasing knowledge of three-dimensional structure of receptors. PMID- 10836126 TI - Laboratory of Chemical Pharmacology, National Heart, Lung, and Blood Institute, NIH: a short history. AB - The Laboratory of Chemical Pharmacology (LCP) began in 1950 as the Section of Pharmacology within the National Heart Institute, the National Institutes of Health. Its first chief was Bernard B. Brodie, considered by many to be one of the fathers of modern pharmacology. Since its inception, LCP has made many significant contributions to the fields of pharmacology and toxicology. LCP was among the first to study (a) the effects of drugs on the turnover of serotonin and norepinephrine in brain and other tissues, (b) the absorption of drugs from the gastrointestinal tract and their passage across the blood-brain barrier, (c) the oxidation and reduction of drugs and other foreign compounds by liver microsomal enzymes (later known as the cytochrome P450 enzymes) and inhibitors and inducers of these enzymes, (d) the formation of toxic chemically reactive metabolites of drugs and other foreign compounds, and (e) mechanisms of immunological responses. Approximately 300 scientists worked in LCP during its existence, and they and their collaborators published more than 1,300 papers. This is a short history of the people who worked in it and of their contributions to biomedical sciences. PMID- 10836127 TI - Chlorinated methanes and liver injury: highlights of the past 50 years. AB - The chlorinated methanes, particularly carbon tetrachloride and chloroform, are classic models of liver injury and have developed into important experimental hepatoxicants over the past 50 years. Hepatocellular steatosis and necrosis are features of the acute lesion. Mitochondria and the endoplasmic reticulum as target sites are discussed. The sympathetic nervous system, hepatic hemodynamic alterations, and role of free radicals and biotransformation are considered. With carbon tetrachloride, lipid peroxidation and covalent binding to hepatic constituents have been dominant themes over the years. Potentiation of chlorinated methane-induced liver injury by alcohols, aliphatic ketones, ketogenic compounds, and the pesticide chlordecone is discussed. A search for explanations for the potentiation phenomenon has led to the discovery of the role of tissue repair in the overall outcome of liver injury. Some final thoughts about future research are also presented. PMID- 10836128 TI - Pharmacokinetic/pharmacodynamic modeling in drug development. AB - We propose a framework for considering the role of pharmacokinetic/pharmacodynamic modeling in drug development and an appraisal of its current and potential impact on that activity. After some introduction, definitions, and background information on drug development, we discuss subject matter models that underlie pharmacokinetic/pharmacodynamic modeling and show how they determine appropriate statistical models. We discuss the broad role modeling can play in drug development, enhancing primarily the "learning" steps, i.e. acquiring the information needed for the label and for planning efficient confirmatory clinical trials. Examples of past applications of modeling to drug development are presented in tabular form, followed by a discussion of some practical issues in application. Modeling will not reach its potential utility until it is manifest as a visible and separate work unit within a drug development program. We suggest that that work unit is the "in numero" study: a protocol-driven exercise designed to extract additional information, and/or answer a specific drug-development question, through an integrated model-based (meta-) analysis of existent raw data, often pooled across separate (clinical) studies. PMID- 10836129 TI - Sequencing the entire genomes of free-living organisms: the foundation of pharmacology in the new millennium. AB - The power and effectiveness of clinical pharmacology are about to be transformed with a speed that earlier in this decade could not have been foreseen even by the most astute visionaries. In the very near future, we will have at our disposal the reference DNA sequence for the entire human genome, estimated to contain approximately 3.5 billion bp. At the same time, the science of whole genome sequencing is fostering the computational science of bioinformatics needed to develop practical applications for pharmacology and toxicology. Indeed, it is likely that pharmacology, toxicology, bioinformatics, and genomics will merge into a new branch of medical science for studying and developing pharmaceuticals from molecule to bedside. PMID- 10836130 TI - High-throughput screening in drug metabolism and pharmacokinetic support of drug discovery. AB - The application of rapid methods currently used for screening discovery drug candidates for metabolism and pharmacokinetic characteristics is discussed. General considerations are given for screening in this context, including the criteria for good screens, the use of counterscreens, the proper sequencing of screens, ambiguity in the interpretation of results, strategies for false positives and negatives, and the special difficulties encountered in drug metabolism and pharmacokinetic screening. Detailed descriptions of the present status of screening are provided for absorption potential, blood-brain barrier penetration, inhibition and induction of cytochrome P450, pharmacokinetics, biotransformation, and computer modeling. Although none of the systems currently employed for drug metabolism and pharmacokinetic screening can be considered truly high-throughput, several of them are rapid enough to be a practical part of the screening paradigm for modern, fast-moving discovery programs. PMID- 10836131 TI - Pharmacogenetics of sulfotransferase. AB - Cytosolic sulfotransferase catalyzes sulfoconjugation of relatively small lipophilic endobiotics and xenobiotics. At least 44 cytosolic sulfotransferases have been identified from mammals, and based on their amino acid sequences, these forms are shown to constitute five different families. In humans, 10 sulfotransferase genes have been identified and shown to localize on at least five different chromosomes. The enzymatic properties characterized in the recombinant forms indicate the association of their substrate specificity with metabolisms of such nonpeptide hormones as estrogen, corticoid, and thyroxine, although most forms are also active on the sulfation of various xenobiotics. Genetic polymorphisms are observed on such human sulfotransferases as ST1A2, ST1A3, and ST2A3. PMID- 10836132 TI - Drug discovery in the next millennium. AB - Selection and validation of novel molecular targets have become of paramount importance in light of the plethora of new potential therapeutic drug targets that have emerged from human gene sequencing. In response to this revolution within the pharmaceutical industry, the development of high-throughput methods in both biology and chemistry has been necessitated. This review addresses these technological advances as well as several new areas that have been created by necessity to deal with this new paradigm, such as bioinformatics, cheminformatics, and functional genomics. With many of these key components of future drug discovery now in place, it is possible to map out a critical path for this process that will be used into the new millennium. PMID- 10836133 TI - The impact of genomics on drug discovery. AB - High-throughput gene sequencing has revolutionized the process used to identify novel molecular targets for drug discovery. Thousands of new gene sequences have been generated but only a limited number of these can be converted into validated targets likely to be involved in disease. We describe here some of the approaches used at SmithKline Beecham to select and validate novel targets. These include the identification of selective tissue gene product expression, such as for cathepsin K, a novel osteoclast-specific cysteine protease. We also describe the discovery and functional characterization of novel members of the G-protein coupled receptor superfamily and their pairing with natural ligands. Lastly, we discuss the promises of gene microarrays and proteomics, developing technologies that allow the parallel analyses of tissue expression patterns of thousands of genes or proteins, respectively. PMID- 10836134 TI - Simulation of clinical trials. AB - Computer simulation of clinical trials has evolved over the past two decades from a simple instructive game to "full" simulation models yielding pharmacologically sound, realistic trial outcomes. The need to make drug development more efficient and informative and the awareness that many industries make extensive use of simulation in product development have advanced considerably the use of simulation of clinical trials in pharmaceutical product development over the past decade. The structural and stochastic components of trial simulation models are explained as a prelude to a listing of representative simulation projects, reflecting investigative applications of statistical methods, trial design comparisons, and full simulation of new drugs being developed. Lessons learned from these projects are reviewed in the context of their current impact and potential for influencing the future of drug development. PMID- 10836135 TI - The regulator of G protein signaling family. AB - Regulator of G protein signaling (RGS) proteins are responsible for the rapid turnoff of G protein-coupled receptor signaling pathways. The major mechanism whereby RGS proteins negatively regulate G proteins is via the GTPase activating protein activity of their RGS domain. Structural and mutational analyses have characterized the RGS/G alpha interaction in detail, explaining the molecular mechanisms of the GTPase activating protein activity of RGS proteins. More than 20 RGS proteins have been isolated, and there are indications that specific RGS proteins regulate specific G protein-coupled receptor pathways. This specificity is probably created by a combination of cell type-specific expression, tissue distribution, intracellular localization, posttranslational modifications, and domains other than the RGS domain that link them to other signaling pathways. In this review we discuss what has been learned so far about the role of RGS proteins in regulating G protein-coupled receptor signaling and point out areas that may be fruitful for future research. PMID- 10836136 TI - Parallel array and mixture-based synthetic combinatorial chemistry: tools for the next millennium. AB - Technological advances continue to be a central driving force in the acceleration of the drug discovery process. Combinatorial chemistry methods, developed over the past 15 years, represent a paradigm shift in drug discovery. Initially viewed as a curiosity by the pharmaceutical industry, combinatorial chemistry is now recognized as an essential tool that decreases the time of discovery and increases the throughput of chemical screening by as much as 1000-fold. The use of parallel array synthesis approaches and mixture-based combinatorial libraries for drug discovery is reviewed. PMID- 10836137 TI - Pharmacology of selectin inhibitors in ischemia/reperfusion states. AB - Recently, the selectin family of glycoprotein adhesion molecules (P-selectin, E selectin, and L-selectin) has been implicated in the pathogenesis of a number of inflammatory disease states. The selectins modulate the early adhesive interactions between circulating neutrophils and the endothelium. Both P-selectin and E-selectin can be expressed on the surface of endothelial cells following stimulation by a number of inflammatory mediators. In contrast, L-selectin is constitutively expressed on the surface of neutrophils at very high levels. In addition, neutrophils also express ligands for the endothelial selectins, including the carbohydrate sialyl Lewis(x) and the high-affinity ligand P selectin glycoprotein ligand 1, which facilitate neutrophil-endothelial interactions. Selectins have been extensively investigated in ischemia/reperfusion injury states. The study of selectin involvement in ischemia/reperfusion injury has been facilitated by the development of highly specific selectin antagonists, including monoclonal antibodies, carbohydrates, small molecule inhibitors, and soluble forms of P-selectin glycoprotein ligand 1. This article reviews the results of current studies of selectin antagonists in experimental models of ischemia/reperfusion injury. PMID- 10836138 TI - A novel means of drug delivery: myoblast-mediated gene therapy and regulatable retroviral vectors. AB - A potentially powerful approach to drug delivery in the treatment of disease involves the use of cells to introduce genes encoding therapeutic proteins into the body. Candidate genes for delivery include those encoding secreted factors that could have broad applications ranging from treatment of inherited single gene deficiencies to acquired disorders of the vasculature or cancer. Myoblasts, the proliferative cell type of skeletal muscle tissues, are potent tools for stable delivery of a gene of interest into the body, as they become an integral part of the muscle into which they are injected, in close proximity to the circulation. The recent development of improved tetracycline-inducible retroviral vectors allows for fine control of recombinant gene expression levels. The combination of ex vivo gene transfer using myoblasts and regulatable retroviral vectors provides a powerful toolbox with which to develop gene therapies for a number of human diseases. PMID- 10836139 TI - 5-ht6 receptors as emerging targets for drug discovery. AB - 5-ht6 receptors are the latest serotonin receptors to be identified by molecular cloning. Their high affinity for a wide range of drugs used in psychiatry, coupled with their intriguing distribution in the brain, has stimulated significant interest. Antisense oligonucleotides, antipeptide antibodies, selective radioligands, knockout mice, and selective antagonists of the 5-ht6 receptor have recently become available. Surprisingly, 5-ht6 receptors appear to regulate cholinergic neurotransmission in the brain, rather than the expected interaction as modulators of dopaminergic transmission. This interaction predicts a possible role for 5-ht6 receptor antagonists in the treatment of learning and memory disorders. Furthermore, polymorphisms in the sequence of the 5-ht6 receptor gene may provide a genetic tool to further our understanding of the differential responses of patients to antipsychotic medications. PMID- 10836140 TI - The impact of genomics-based technologies on drug safety evaluation. AB - Determining the potential toxicity of compounds early in the drug discovery process can be extremely beneficial in terms of both time and money conservation. Because of the speed of modern chemical synthesis and screening, to accurately evaluate the large number of compounds being produced, toxicology assays must have both high-fidelity and high-throughput capabilities. In addition, assays must be performed using limited amounts of compound. In the past decade, several new and innovative techniques have been developed that not only allow for high throughput screening but can also provide detailed information concerning the molecular mechanisms behind toxic effects. Techniques such as hybridization microarrays, real-time polymerase chain reaction, and large-scale sequencing are some of the methods that have been or are starting to be used routinely in pharmaceutical companies. This review examines the contributions of these and related techniques toward toxicity evaluation of potential drug candidates and their future role in the discovery of new therapeutics. PMID- 10836141 TI - Mitochondrial targets of drug toxicity. AB - Mitochondria have long been recognized as the generators of energy for the cell. Like any other power source, however, mitochondria are highly vulnerable to inhibition or uncoupling of the energy harnessing process and run a high risk for catastrophic damage to the cell. The exquisite structural and functional characteristics of mitochondria provide a number of primary targets for xenobiotic-induced bioenergetic failure. They also provide opportunities for selective delivery of drugs to the mitochondrion. In light of the large number of natural, commercial, pharmaceutical, and environmental chemicals that manifest their toxicity by interfering with mitochondrial bioenergetics, it is important to understand the underlying mechanisms. The significance is further underscored by the recent identification of bioenergetic control points for cell replication and differentiation and the realization that mitochondria play a determinant role in cell signaling and apoptotic modes of cell death. PMID- 10836142 TI - Molecular mechanisms and regulation of opioid receptor signaling. AB - Cloning of multiple opioid receptors has presented opportunities to investigate the mechanisms of multiple opioid receptor signaling and the regulation of these signals. The subsequent identification of receptor gene structures has also provided opportunities to study the regulation of receptor gene expression and to manipulate the concentration of the gene products in vivo. Thus, in the current review, we examine recent advances in the delineation basis for the multiple opioid receptor signaling, and their regulation at multiple levels. We discuss the use of receptor knockout animals to investigate the function and the pharmacology of these multiple opioid receptors. The reasons and basis for the multiple opioid receptor are addressed. PMID- 10836143 TI - Nicotinic receptors at the amino acid level. AB - nAChRs are pentameric transmembrane proteins into the superfamily of ligand-gated ion channels that includes the 5HT3, glycine, GABAA, and GABAC receptors. Electron microscopy, affinity labeling, and mutagenesis experiments, together with secondary structure predictions and measurements, suggest an all-beta folding of the N-terminal extracellular domain, with the connecting loops contributing to the ACh binding pocket and to the subunit interfaces that mediate the allosteric transitions between conformational states. The ion channel consists of two distinct elements symmetrically organized along the fivefold axis of the molecule: a barrel of five M2 helices, and on the cytoplasmic side five loops contributing to the selectivity filter. The allosteric transitions of the protein underlying the physiological ACh-evoked activation and desensitization possibly involve rigid body motion of the extracellular domain of each subunit, linked to a global reorganization of the transmembrane domain responsible for channel gating. PMID- 10836144 TI - The role of Rho in G protein-coupled receptor signal transduction. AB - Low molecular weight G proteins of the Rho subfamily are regulators of actin cytoskeletal organization. In contrast to the heterotrimeric G proteins, the small GTPases are not directly activated through ligand binding to G protein coupled receptors (GPCRs). However, a subset of GPCRs, including those for lysophosphatidic acid and thrombin, induce stress fibers, focal adhesions, and cell rounding through Rho-dependent pathways. C3 exoenzyme has been a useful tool for demonstrating Rho involvement in these and other responses, including Ca2+ sensitization of smooth muscle contraction, cell migration, transformation, and serum response element-mediated gene expression. Most of the GPCRs that induce Rho-dependent responses can activate Gq, but this is not a sufficient signal. Recent data demonstrate that G alpha 12/13 can induce Rho-dependent responses. Furthermore, G alpha 12/13 can bind and activate Rho-specific guanine nucleotide exchange factors, providing a mechanism by which GPCRs that couple to G alpha 12/13 could activate Rho and its downstream responses. PMID- 10836145 TI - Central role of peroxisome proliferator-activated receptors in the actions of peroxisome proliferators. AB - Peroxisome proliferators (PPs) are a large class of structurally dissimilar chemicals that have diverse effects in rodents and humans. Most, if not all, of the diverse effects of PPs are mediated by three members of the nuclear receptor superfamily called peroxisome proliferator-activated receptors (PPARs). In this review, we define the molecular mechanisms of PPs, including PPAR binding specificity, alteration of gene expression through binding to DNA response elements, and cross talk with other signaling pathways. We discuss the roles of PPARs in growth promotion in rodent hepatocarcinogenesis and potential therapeutic effects, including suppression of cancer growth and inflammation. PMID- 10836146 TI - The PAS superfamily: sensors of environmental and developmental signals. AB - Over the past decade, PAS domains have been identified in dozens of signal transduction molecules and various forms have been found in animals, plants, and prokaryotes. In this review, we summarize this rapidly expanding research area by providing a detailed description of three signal transduction pathways that utilize PAS protein heterodimers to drive their transcriptional output. It is hoped that these model pathways can provide a framework for use in understanding the biology of the less well-understood members of this emerging superfamily, as well as of those to be characterized in the days to come. We use this review to develop the idea that most eukaryotic PAS proteins can be classified by functional similarities, as well as by predicted phylogenetic relationships. We focus on the alpha-class proteins, which often act as sensors of environmental signals, and the beta-class proteins, which typically act as broad-spectrum partners that target these heterodimers to their genomic targets. PMID- 10836147 TI - Pharmacology of cloned P2X receptors. AB - There are seven P2X receptor cDNAs currently known. Six homomeric (P2X1, P2X2, P2X3, P2X4, P2X5, P2X7) and three heteromeric (P2X2/P2X3, P2X4/P2X6, P2X1/P2X5) P2X receptor channels have been characterized in heterologous expression systems. Homomeric P2X1 and P2X3 receptors are readily distinguishable by their rapid desensitization, the agonist action of alpha beta methyleneATP, and the block by 2',3'-O-(2,4,6-trinitrophenyl)-ATP. P2X2 receptors are unique among homomeric forms in their potentiation by low pH. Homomeric P2X4 receptors are much less sensitive to antagonism by suramin and pyridoxal 5-phosphate-6-azo-2',4' disulfonic acid. Homomeric P2X7 receptors are the only form in which 2',3'-O-(4 benzoylbenzoyl)-ATP is more potent than ATP. The heteromeric P2X2/P2X3 receptor resembles P2X2 in slow desensitization kinetics and potentiation by low pH and is similar to P2X3 with respect to agonism by alpha beta methyleneATP and block by 2',3'-O-(2,4,6-trinitrophenyl)-ATP. Other agonists, antagonists, and ions that can be used to differentiate among the receptors are discussed. PMID- 10836148 TI - Human UDP-glucuronosyltransferases: metabolism, expression, and disease. AB - In vertebrates, the glucuronidation of small lipophilic agents is catalyzed by the endoplasmic reticulum UDP-glucuronosyltransferases (UGTs). This metabolic pathway leads to the formation of water-soluble metabolites originating from normal dietary processes, cellular catabolism, or exposure to drugs and xenobiotics. This classic detoxification process, which led to the discovery nearly 50 years ago of the cosubstrate UDP-glucuronic acid (19), is now known to be carried out by 15 human UGTs. Characterization of the individual gene products using cDNA expression experiments has led to the identification of over 350 individual compounds that serve as substrates for this superfamily of proteins. This data, coupled with the introduction of sophisticated RNA detection techniques designed to elucidate patterns of gene expression of the UGT superfamily in human liver and extrahepatic tissues of the gastrointestinal tract, has aided in understanding the contribution of glucuronidation toward epithelial first-pass metabolism. In addition, characterization of the UGT1A locus and genetic studies directed at understanding the role of bilirubin glucuronidation and the biochemical basis of the clinical symptoms found in unconjugated hyperbilirubinemia have uncovered the structural gene polymorphisms associated with Crigler-Najjar's and Gilbert's syndrome. The role of the UGTs in metabolism and different disease states in humans is the topic of this review. PMID- 10836149 TI - 14-3-3 proteins: structure, function, and regulation. AB - The 14-3-3 proteins are a family of conserved regulatory molecules expressed in all eukaryotic cells. A striking feature of the 14-3-3 proteins is their ability to bind a multitude of functionally diverse signaling proteins, including kinases, phosphatases, and transmembrane receptors. This plethora of interacting proteins allows 14-3-3 to play important roles in a wide range of vital regulatory processes, such as mitogenic signal transduction, apoptotic cell death, and cell cycle control. In this review, we examine the structural basis for 14-3-3-ligand interactions, proposed functions of 14-3-3 in various signaling pathways, and emerging views of mechanisms that regulate 14-3-3 actions. PMID- 10836150 TI - Dual protease inhibitor therapy in HIV-infected patients: pharmacologic rationale and clinical benefits. AB - HIV protease inhibitors, as components of combination antiretroviral drug regimens, have substantially reduced the morbidity and mortality associated with HIV infection. They selectively block the action of the virus-encoded protease and stop the virus from replicating. In general, these drugs have poor systemic bioavailability and must be dosed with respect to meals for optimal absorption. Protease inhibitor-containing regimens require ingestion of a large number of capsules, are costly, and produce or are susceptible to metabolic drug interactions. Simultaneous administration of two protease inhibitors takes advantage of beneficial pharmacokinetic interactions and may circumvent many of the drugs' undesirable pharmacologic properties. For example, ritonavir increases saquinavir concentrations at steady state by up to 30-fold, allowing reduction of saquinavir dose and dosing frequency. Ritonavir decreases the systemic clearance of indinavir and overcomes the deleterious effect of food on indinavir bioavailability. These benefits reflect inhibition of presystemic clearance and first-pass metabolism, as well as inhibition of systemic clearance mediated by hepatic cytochrome P450 3A4. Several dual protease inhibitor combination regimens have shown great promise in clinical trials and are now recommended as components of salvage therapy for HIV-infected patients. PMID- 10836151 TI - Current concepts on pulmonary host defense mechanisms in children. AB - The respiratory tract is exposed continuously to noxious agents, microbial organisms, particles, and allergens. It has therefore evolved both innate and specific defense mechanisms. The innate host defense mechanisms include components such as collectins, beta-defensins, lactoferrin, and complement, all of which have an important role in modulating the immune response. Immune protection of the lungs by specific antibody is reviewed. The airways are protected by alveolar macrophages, neutrophils, and lymphocytes, and their origins, regulation, functions, and antimicrobial activity are summarized. Antimicrobial peptides and immune-modulating peptides are likely to have a significant therapeutic role for infection and inflammation in the respiratory tract. PMID- 10836152 TI - Current concepts on imaging of thoracic vascular abnormalities. AB - Pediatric thoracic vascular abnormalities include many anatomic variants and pathologic conditions. Although some vascular variants are inconsequential and are discovered incidentally on imaging studies, several anomalies have profound effects on the airway and pulmonary parenchyma. Imaging plays a key role in evaluating the chest and its vasculature. The chest radiograph is usually the first screening study performed during the course of evaluation for a vascular abnormality. Cross-sectional imaging with CT and magnetic resonance imaging provides precise anatomic information and has in most cases replaced invasive diagnostic angiographic procedures. We describe common thoracic vascular abnormalities that occur in children and the imaging techniques currently used in their evaluation. PMID- 10836153 TI - Current concepts of respiratory complications of neuromuscular disease in children. AB - Chronic neuromuscular diseases affect the respiratory muscles in varying patterns and degrees. As a result, patients with these disorders develop restrictive pulmonary disease, ineffective cough, atelectasis and pneumonia, and chronic respiratory insufficiency leading to respiratory failure. Therapeutic strategies are evolving to augment cough and airway clearance, improve lung volumes, and support the patient with progressive ventilatory failure. These techniques have improved longevity and quality of life for many patients with neuromuscular disease. PMID- 10836154 TI - Sleep-disordered breathing in children. AB - Sleep-disordered breathing, particularly the obstructive sleep apnea syndrome, is common during childhood. Complications include neurocognitive deficits, growth failure, and pulmonary hypertension. Nevertheless, sleep-disordered breathing is often unrecognized in children. New syndromes, such as the upper airway resistance syndrome, have recently been described. Polysomnography is invaluable for the evaluation of sleep-disordered breathing. Advances in diagnosis and management can alleviate much of the morbidity previously associated with sleep related respiratory disorders. PMID- 10836155 TI - All that wheezes.... PMID- 10836156 TI - Problems associated with intravenous fluid administration in children: do we have the right solutions? AB - Edema, both in its cellular and tissue form, is a frequent complication in critically ill patients. In health, a balance is struck between intracellular and extracellular osmotic forces and interstitial and intravascular oncotic forces, which in turn governs fluid distribution between the intracellular, extracellular and interstitial fluid compartments. The use of hypo- and hypernotic electrolyte solutions has major effects on brain cells, which can be detrimental or beneficial. The controversy over the use of colloids versus crystalloid solutions in critically ill patients remains unresolved. PMID- 10836157 TI - Neurological critical care. AB - Of utmost importance in the practice of neurological critical care is the treatment of cerebral edema, when possible, and the control of life-threatening seizures. In this regard, severe traumatic head injury and refractory status epilepticus are useful clinical therapeutic paradigms. Evidence-based treatment established for these conditions has, by necessity, a wider application to other much less frequent causes of coma and acute neurological illness managed in the intensive therapy unit. Therefore, this review of pediatric neurocritical care literature in 1999 highlights central clinical reports of the medical management of severe traumatic brain injury, the benzodiazepines used in the treatment of status epilepticus, and the emerging or recently appreciated encephalopathies occurring in children. PMID- 10836158 TI - Neonatal acute respiratory failure. AB - Acute respiratory failure is the most common problem seen in the preterm and term infants admitted to neonatal intensive care units. In preterm infants, the most common cause of acute respiratory failure is respiratory distress syndrome caused by surfactant deficiency. Acute respiratory failure in term and near term infants is usually a result of meconium aspiration syndrome, sepsis, pulmonary hypoplasia, and primary pulmonary hypertension of the newborn. The response to various methods of treatment may vary, depending on the severity of respiratory failure and the cause of the acute respiratory failure. We reviewed the evidence for efficacy and current utilization of newer treatment modalities, including exogenous surfactant administration, high frequency ventilation, inhaled nitric oxide therapy, antenatal steroids for the prevention of respiratory distress syndrome, and use of postnatal steroids for the prevention of chronic lung disease. PMID- 10836159 TI - Acute respiratory failure in children. AB - Recent studies in the treatment of acute respiratory failure in children have been targeted at reducing ventilator-induced lung injury, providing treatment adjuncts to mechanical ventilation, and assessing innovative therapies directed at immunomodulation. Ventilator-associated lung injury has been demonstrated in animal models during the delivery of moderate-to-large tidal volumes and has also been described in adult populations. Subsequently, a significant survival benefit of a low tidal volume, high positive end expiratory pressure strategy on the ventilatory was found in adults. Investigation of the effects of inhaled nitric oxide in acute respiratory failure patients continues to show transient improvements in oxygenation, but no evidence of improved outcomes. The use of intratracheal surfactant within 24 hours of intubation in pediatric respiratory failure may be beneficial in reducing the days of mechanical ventilation. Neutrophil oxidative damage has been demonstrated, but therapies directed at decreasing neutrophil adherence have failed to demonstrate improvements. Enteral anti-inflammatory and antioxidant therapy may be promising, because these modalities have been shown to improve a number of surrogate outcomes in patients with respiratory failure. The use of corticosteroids in the late stages of lung injury has also recently been shown to have promise. PMID- 10836160 TI - Pediatric cardiac intensive care: current state of the art and beyond the millennium. AB - Pediatric cardiac intensive care has emerged as a distinct clinical entity to meet the unique needs of pediatric patients with congenital and acquired heart disease. This new subspecialty demands expertise and experience in the pediatric subspecialties of cardiology, intensive care, cardiac surgery, cardiac anesthesia, neonatology, and others. Ten recent developments will have an impact on pediatric cardiac intensive care for the coming decades: 1) emergence of new patient populations; 2) new clinical methodologies in the treatment of pulmonary hypertension; 3) innovations in techniques of respiratory support; 4) expanding research of single ventricle physiology; 5) advances in the treatment of heart failure; 6) improved noninvasive imaging; 7) new directions in interventional cardiac catheterization; 8) new techniques in pediatric cardiac surgery; 9) use of computer technology and intensive care monitoring; and 10) appreciation for global economics of intensive care. Finally, a multidisciplinary approach with a team esprit de corps remains vital to a successful pediatric cardiac intensive care program. PMID- 10836161 TI - Medical management of gastroesophageal reflux. AB - Gastroesophageal reflux (GER) is a common problem which can manifest as vomiting, failure to thrive, recurrent pneumonias, asthma, sinusitus, or subglottic stenosis. The medical management plan should be individualized. A "happy spitter" who has no complications of GER may respond well to conservative management, including positioning and thickening of feedings. A child with complications may require treatment with H-2 antagonists or proton pump inhibitors in conjunction with prokinetic agents. Children with gastrointestinal symptoms suggestive of GER who do not respond to antireflux management may need to be treated for eosinophilic esophagitis. Recent studies that assess the effect of medications on recognized complications of GER are reviewed. PMID- 10836162 TI - Periodic fever, aphthous stomatitis, pharyngitis, adenitis: a clinical review of a new syndrome. AB - Periodic fevers (fevers that occur predictably at fixed intervals) are unusual in infants and children. The classic periodic fever syndrome is cyclic neutropenia (neutropenia followed by infections and fever that recur every 21 days). A new periodic fever syndrome PFAPA (periodic fever, aphthous stomatitis, pharyngitis, and cervical adenitis) has been characterized over the past decade. PFAPA is defined clinically, because specific laboratory abnormalities have not been found. The clinical characteristic of PFAPA is high fevers (usually 40.0 degrees C to 40.6 degrees C) recurring at fixed intervals every 2 to 8 weeks. The fevers last for about 4 days, then resolve spontaneously. Associated with the fevers are aphthous stomatitis in 70% of patients, pharyngitis in 72% of patients, and cervical adenitis in 88% of patients. PFAPA is not familial and begins before the age of 5 years. An episode of PFAPA can be aborted with one or two small doses of prednisone. The episodes of PFAPA may last for years and the patient is well between episodes. The cause of PFAPA is unknown and there are no reported sequelae. PMID- 10836164 TI - Acute complications of Epstein-Barr virus infectious mononucleosis. AB - Infectious mononucleosis caused by Epstein-Barr virus (EBV) usually resolves over a period of weeks or months without sequelae but may occasionally be complicated by a wide variety of neurologic, hematologic, hepatic, respiratory, and psychological complications. The strength of association of EBV with many of these complications remains based on scattered case reports, often using unsophisticated diagnostic tests, and the evidence for causation in many instances is unconvincing. There is little benefit of antiviral treatment of uncomplicated or complicated infectious mononucleosis. Corticosteroids may have a role in hastening resolution of some complications, especially upper airway obstruction and possibly immune-mediated anemia and thrombocytopenia, but should be used judiciously. PMID- 10836163 TI - Respiratory syncytial virus vaccine development. AB - Respiratory syncytial virus is the major respiratory pathogen of infants and children worldwide. Currently, there is no effective vaccine to protect against respiratory syncytial virus infection. Immunoprophylaxis with hyperimmune globulin or with a humanized monoclonal antibody is expensive, limited to children with underlying disease, and not practical for general use. Antiviral therapy is controversial and of limited effectiveness. New approaches to the development of a vaccine for respiratory syncytial virus infection are promising. Several subunit vaccines and live attenuated virus vaccines are immunogenic and safe in children and adults. This review focuses on potential vaccine candidates and the challenges these candidate vaccines must overcome. PMID- 10836165 TI - Human herpesvirus-6 and -7 infections in children: agents of roseola and other syndromes. AB - Human herpesvirus-6 (HHV-6) and -7 (HHV-7) infections typically are silent or manifested as mild febrile illnesses including classic roseola. In addition, case reports and epidemiologic data support the rare occurrence of HHV-6 encephalitis in immunocompromised as well as immunocompetent subjects. Although many other diseases have been putatively associated with HHV-6 or HHV-7, these associations are not well documented due to small numbers, use of tests incapable of distinguishing latent from replicating virus, potential virus cross-reactivity, or contradictory results. Further careful studies are needed to confirm these disease associations. Laboratory tests for diagnosing active HHV-6 and HHV-7 infections include virus culture, antigen detection, and polymerase chain reaction of cell-free biologic fluid. Although HHV-6 and HHV-7 are inhibited by several antiviral drugs in the laboratory, including ganciclovir and foscarnet, no clinical trials have assessed their benefit. Nevertheless, treatment may be considered for patients with serious HHV-6- or HHV-7-associated disease confirmed with accurate virologic tests. PMID- 10836166 TI - Role of astroviruses in childhood diarrhea. AB - Human astrovirus (HAstV) is a significant cause of acute diarrhea among children, resulting in outbreaks of diarrhea and occasionally in hospitalization. Improved detection methods for eight antigenic types of HAstV and studies assessing the frequency and severity of HAstV diarrhea have further defined the impact of HAstV infections in children. These studies have shown that HAstV infections are clinically milder (diarrhea, vomiting, fever) than rotavirus infections. However, frequent coinfection of HAstV with rotavirus and caliciviruses in childhood diarrhea complicates the epidemiology. Seroprevalence studies have provided evidence that the majority of children are infected by HAstV by 6 years of age. The route of transmission is probably fecal-oral from food or water sources. Recent and planned studies will help to define the epidemiology and in the future lead to prevention strategies, which could include vaccination. PMID- 10836167 TI - Updates in pediatric nutrition. AB - There have been many remarkable advances in pediatric nutrition. Solid scientific evidence now supports certain fundamental assumptions long held in the pediatric community. For example, obesity in children has for some time been believed to have adverse health effects; recent large scale studies now confirm relationships between childhood obesity and specific morbidities. Likewise, the beneficial effects of human breast milk on growth and development have been the focus of recent prospective studies of full term and preterm infants. There has been active research in the area of formula intolerance and allergy, allowing practicing physicians to better counsel parents about dietary choices. Although many health problems are caused by the abundance of high fat and high calorie foods in the average US child's diet, a large number of children remain at risk for hunger in the United States. Other research provides important breakthroughs in our understanding of the impact of pediatric nutrition on lifelong health. Retrospective epidemiological studies have uncovered relationships between prenatal factors and health later in life. These studies have lead to ongoing prospective observational trials that should provide further information about the extent to which certain health factors are determined before birth. In addition, basic science research has revealed previously unknown mechanisms by which essential minerals, such as iron, are transported into the body. In sum, this section reviews exciting new information in the areas of childhood diet quality, obesity, breast milk, formula intolerance, and iron metabolism. PMID- 10836169 TI - Bibliography. Current world literature. Pulmonology. PMID- 10836170 TI - Bibliography. Current world literature. Emergency and critical care pediatrics. PMID- 10836168 TI - Nutrition in adolescent pregnancy. AB - Prevention of unintended adolescent pregnancy is a primary goal of the American Academy of Pediatrics and of many health providers. Nevertheless, many adolescents become pregnant every year in America. Pediatricians therefore should be aware of nutritional recommendations for pregnant adolescents to provide optimal care. The importance of nutrition during pregnancy is here reviewed from a pediatric perspective. Pregnancy, particularly during adolescence, is a time of extreme nutritional risk. The adolescents most likely to become pregnant are often those with inadequate nutritional status and unfavorable socio-economic background. There is increasing evidence of competition for nutrients between the growing pregnant adolescent and her fetus. Also, the prenatal environment has been implicated in the development of obesity, cardiovascular disease, and diabetes in both the mother and her offspring. Many adolescents have poor diet quality and poor knowledge of appropriate nutrition; these habits may not change during pregnancy. Current knowledge and recommendations regarding the intake of energy, calcium, and folate are discussed in detail. PMID- 10836171 TI - Bibliography. Current world literature. Surgery. PMID- 10836172 TI - Bibliography. Current world literature. Infectious diseases and immunization. PMID- 10836173 TI - Demographics of the gay and lesbian population in the United States: evidence from available systematic data sources. AB - This work provides an overview of standard social science data sources that now allow some systematic study of the gay and lesbian population in the United States. For each data source, we consider how sexual orientation can be defined, and we note the potential sample sizes. We give special attention to the important problem of measurement error, especially the extent to which individuals recorded as gay and lesbian are indeed recorded correctly. Our concern is that because gays and lesbians constitute a relatively small fraction of the population, modest measurement problems could lead to serious errors in inference. In examining gays and lesbians in multiple data sets we also achieve a second objective: We provide a set of statistics about this population that is relevant to several current policy debates. PMID- 10836174 TI - Measuring living standards with proxy variables. AB - Very few demographic surveys in developing countries have gathered information on household incomes or consumption expenditures. Researchers interested in living standards therefore have had little alternative but to rely on simple proxy indicators. The properties of these proxies have not been analyzed systematically. We ask what hypotheses can be tested using proxies, and compare these indicators with consumption expenditures per adult, our preferred measure of living standards. We find that the proxies employed in much demographic research are very weak predictors of consumption per adult. Nevertheless, hypothesis tests based on proxies are likely to be powerful enough to warrant consideration. PMID- 10836175 TI - Age reporting among white Americans aged 85+: results of a record linkage study. AB - This study investigates age reporting on the death certificates of older white Americans. We link a sample of death certificates for native-born whites aged 85+ in 1985 to Social Security Administration records and to records of the U.S. censuses of 1900, 1910, and 1920. When ages in these sources are compared, inconsistencies are found to be minimal, even beyond age 95. Results show little distortion and no systematic biases in the reported age distribution of deaths. To explore the effect of age misreporting on old-age mortality, we estimate "corrected" age-specific death rates by the extinct-generation method for the U.S. white cohort born in 1885. With few exceptions, corrected and uncorrected rates in single years differ by less than 3% and are not systematically biased. When we compare corrected rates with those for the same birth cohort in France, Japan, and Sweden, we find that white American mortality at older ages is exceptionally low. PMID- 10836176 TI - Combining registration-system and survey data to estimate birth probabilities. AB - With the widespread availability of event-history data, demographers have increasingly eschewed registration-system data in favor of survey data. We propose instead using survey and registration-system data in combination, via a constrained maximum-likelihood framework for demographic hazard modeling. As an application, we combine panel survey data and birth registration data to estimate annual birth probabilities by parity. The general fertility rate obtained from registration-system data constrains the weighted sum of parity-specific birth probabilities. The variances about the parity-specific birth probabilities are halved when registration-system data are used to constrain the estimates. Other demographic applications are discussed. PMID- 10836177 TI - A note on the measurement of accuracy for subnational demographic estimates. AB - Mean absolute percentage error (MAPE), the measure most often used for evaluating subnational demographic estimates, is not always valid. We describe guidelines for determining when MAPE is valid. Applying them to case study data, we find that MAPE understates accuracy because it is unduly influenced by outliers. To overcome this problem, we calculate a transformed MAPE (MAPE-T) using a modified Box-Cox method. Because MAPE-T is not in the same scale as the untransformed absolute percentage errors, we provide a procedure for calculating MAPE-R, a measure in the same scale as the original observations. We argue that MAPE-R is a more appropriate summary measure of average absolute percentage error when the guidelines indicate that MAPE is not valid. PMID- 10836178 TI - Child support and the postdivorce economic well-being of mothers, fathers, and children. AB - This article provides national estimates of the current and potential impact of private child support transfers on the economic well-being of custodial and noncustodial families following marital dissolution. Mothers and children fare dramatically worse than fathers after marital dissolution; these differences, however, would be much more pronounced in the absence of private child support. Simulations of four existing child support guidelines show that substantial increases in economic well-being among mother-custody families are possible within the structure of the existing child support system, with minimal impact on poverty among nonresident fathers. Under all of these guidelines, however, custodial-mother families would continue to fare substantially worse than nonresident fathers. PMID- 10836179 TI - Regional analysis of divorce in China since 1980. AB - This paper estimates and discusses divorce measures in China and its 30 provinces since 1980. The propensity for divorce in China increased 42% between 1982 and 1990. Substantial increases in divorce were observed in all provinces except Xinjiang and Shanxi. We found a fairly strong regional pattern of divorce in China: Divorce rates were lowest on the east coast and in eastern areas, highest in the northwest and the northeast, and moderate in the middle and southern parts of the country. Descriptive explanations of trends and regional variations are presented. PMID- 10836180 TI - Social security, economic growth, and the rise in elderly widows' independence in the twentieth century. AB - The percentage of elderly widows living alone rose from 18% in 1940 to 62% in 1990, while the percentage living with adult children declined from 59% to 20%. This study finds that income growth, particularly increased Social Security benefits, was the single most important determinant of living arrangements, accounting for nearly one-half of the increase in independent living. Unlike researchers in earlier studies, we find no evidence that the effect of income become stronger over the period. Changes in age, race, immigrant status, schooling, and completed fertility explain a relatively small share of the changes in living arrangements. PMID- 10836181 TI - How does POSSLQ measure up? Historical estimates of cohabitation. AB - We use March Current Population Survey (CPS) data from 1977 to 1997 to produce a new historical series of indirect cohabitation prevalence estimates. We compare our new estimates with those produced by the traditional method and evaluate the new estimates. We then compare the indirect estimates with the new direct estimates to investigate whether biases exist in the indirect estimates. Our findings indicate that the traditional indirect method of estimating cohabitation prevalence underestimates cohabitors in different subpopulations, especially among those with children. We also find that the new indirect measure produces relatively unbiased estimates of cohabitors' characteristics. PMID- 10836182 TI - Commentary on "A methodology for identifying married couples in Medicare data". AB - Authors of an article appearing recently in Demography propose a retrospective method to identify husband-wife pairs in administrative records. Although their premise is correct, they over-state the power of their methodology. The error stems from a misunderstanding of how persons entitled to both primary and secondary social security benefits are classified in program data. A retrospective look four years after the reference data shows that the number of pairs is increased by less than 10%, not by more than 50%, as the authors claim. PMID- 10836183 TI - Refinements of a methodology for detecting married couples in the Medicare data. PMID- 10836184 TI - Acute gastrointestinal bleeding in the intensive care unit. The intensivist's perspective. AB - Gastrointestinal (GI) hemorrhage is a common and potentially lethal medical emergency that is a common cause for intensive care unit admission. The intensivist plays an important role as a member of the medical team managing the patient with GI bleeding who is at high risk because of severe bleeding, comorbidity, or the presence of endoscopic stigmata of recent hemorrhage. This article presents the intensivist's approach to GI hemorrhage in initial patient assessment, triage, resuscitation, specialist consultation, diagnostic evaluation, and medical therapy. This article focuses on types of GI bleeding of particular concern to the intensivist, including esophageal variceal bleeding, stress-related GI bleeding, and GI bleeding associated with myocardial infarcation. PMID- 10836185 TI - Acute gastrointestinal bleeding in the intensive care unit. The gastroenterologist's perspective. AB - Although SRES-associated hemorrhage previously constituted a significant cause of bleeding in the ICU, improvements in ICU management and the institution of prophylactic measures in high-risk patients have significantly reduced SRES associated hemorrhage since the 1980s. Antacids, H2-receptor antagonists, and sucralfate have been shown to be effective in preventing clinically significant bleeding resulting from SRES, particularly when the intragastric pH is maintained at greater than 4. A selective approach should be adopted in SRES prophylaxis: Patients on mechanical ventilation, with coagulopathy, or with two of the other known risk factors should receive prophylaxis. Although the drug of choice depends to some extent on local preferences, an H2-receptor antagonist by continuous intravenous infusion may represent the best option. No pharmacologic therapy is of proven value once hemorrhage begins, but the current interventional techniques are effective in controlling hemorrhage. Gastrointestinal bleeding from NOMV has become less common with improvements in the hemodynamic monitoring of critically ill patients, but this disease must always be considered when lower gastrointestinal bleeding occurs in the context of relative hypoperfusion. For SRES and NOMV, treatment of the underlying disease or diseases is the optimal route to prevention. PMID- 10836186 TI - Esophageal varices. I. Clinical presentation, medical therapy, and endoscopic therapy. AB - The last half century has witnessed great advances in the understanding of the pathogenesis and natural history of portal hypertension in cirrhotics. Several pharmacologic and endoscopic techniques have been developed for the treatment of portal hypertension. The use of these agents in a given patient must be based on an understanding of the stage in the natural history of the disease and the relative efficacy and safety of the available treatment options. PMID- 10836187 TI - Esophageal varices. II. TIPS (transjugular intrahepatic portosystemic shunt) and surgical therapy. AB - The role of surgery in the prevention and treatment of variceal hemorrhage is reviewed. Types of available surgery, their physiologic basis, and literature supporting their use are discussed in the context of the natural history of variceal hemorrhage. The evolution of transjugular intrahepatic portosystemic shunt (TIPS) as a treatment modality for variceal hemorrhage is reviewed. The effects of TIPS on portal and systemic hemodynamics and clinical usefulness in the management of variceal hemorrhage are discussed. A treatment algorithm for the integrated use of the various treatments is provided. PMID- 10836188 TI - Gastrointestinal bleeding associated with myocardial infarction. AB - Gastrointestinal (GI) bleeding associated with myocardial infarcation (MI) often presents as a distinct syndrome that differs from either disease alone. MI is frequently overlooked in the setting of severe GI bleeding because the symptoms and signs of MI are frequently overshadowed by the severe bleeding. GI bleeding, particularly when massive, may precipitate MI from hypovolemia, hemodynamic compromise, and myocardial hypoperfusion. Esophagogastroduodenoscopy is safe in relatively clinically stable patients after MI and is indicated to evaluate significant upper GI bleeding. PMID- 10836189 TI - Clinical scoring systems for determining the prognosis of gastrointestinal bleeding. AB - The prognosis of GI bleeding depends upon many factors. Patients should be evaluated carefully for risk factors. To avoid complications from GI bleeding, triage should be performed promptly after patient presentation. The history and physical examination should emphasize analysis of risk factors for severe GI bleeding and mortality. Factors that increase the morbidity and mortality include: age greater than 60 years; underlying comorbidity such as pulmonary diseases, liver diseases, renal diseases, encephalopathy, or cancer; physiologic stress from major surgery, trauma, or sepsis; coexisting disease in three organ systems; low hematocrit; melena or hematochezia; and prolonged prothrombin time. Hospitalized patients who require more than five units of packed erythrocytes transfusion or who develop hypotension or hypovolemic shock are more likely to need surgery. Patients with a high APACHE II score, the presence of esophageal varices, active bleeding, or other endoscopic stigmata of recent hemorrhage are more likely to rebleed and undergo surgery. The proliferation of multivariable prognostic scales, as described herein, provides ample evidence that the goal of developing a single comprehensive multivariable scale to accurately assess severity of disease and to determine prognosis of GI bleeding is still not achieved. Yet significant progress has occurred in this field, leading to the hope of developing a universally applicable multivariable scale. PMID- 10836190 TI - Therapeutic endoscopy for nonvariceal gastrointestinal bleeding. AB - This article reviews the role of therapeutic endoscopy in the diagnosis and treatment of nonvariceal upper and lower gastrointestinal (GI) hemorrhage. The initial approach to patients with GI bleeding is reviewed. Endoscopic treatment of various stigmata of recent peptic ulcer hemorrhage is discussed in detail. Management of less common causes of nonvariceal bleeding, such as Dieulafoy's lesions, Mallory-Weiss tears, angiomas, and bleeding colonic diverticula is described. Recommendations for endoscopic techniques are based on the results of UCLA-CURE hemostasis studies. PMID- 10836191 TI - Radiology in the diagnosis and therapy of gastrointestinal bleeding. AB - Major breakthroughs in catheter and guidewire design as well as improvements in angiographic x-ray equipment currently allow interventional radiologists to diagnose massive life-threatening upper and lower GI hemorrhage and to stop the bleeding safely and effectively using superselective catheterization and microcoil embolization. For chronic or recurrent GI bleeding, when endoscopy is unrevealing or equivocal, barium studies, CT scanning, nuclear medicine studies, and angiography can help determine the cause of bleeding. A multidisciplinary approach, including the gastroenterologist, radiologist, and surgeon, is extremely helpful in managing GI bleeding, particularly in high-risk patients or patients presenting as diagnostic dilemmas. PMID- 10836192 TI - Fifty landmark discoveries in gastroenterology during the past 50 years. A brief history of modern gastroenterology at the millennium: Part II. Gastrointestinal motility, nutrition, and diseases of the lower gastrointestinal tract, liver, and pancreas. AB - During the last half century, many outstanding discoveries have revolutionized the clinical practice and science of gastroenterology. Although the scientific results are widely disseminated, the discoverers have received inadequate recognition and the history of their discoveries is largely unstudied and unknown. At the millennium, a committee selected 50 landmark discoveries in gastroenterology during the past 50 years. A brief history of each landmark discovery is presented. Part I was presented in the previous issue of Gastroenterology Clinics of North America. Part II presents landmark discoveries in gastrointerintal (GI) motility, clinical trials, nutrition, and diseases of the lower GI tract, liver, biliary tree, and pancreas. PMID- 10836194 TI - The importance of user compliance on the effectiveness of natural family planning programs. AB - Nowadays, there is an increasing interest in natural family planning methods. The biological basis for the application of natural family planning methods is the recognition of ovulation and, more extensively, of the fertile period. Several studies in the past decade have shown the efficacy of these methods and that the main cause of failure was either a conscious departure from the rules of the method or erroneous application of the method. Another problem affecting natural family planning that has been highlighted is the relatively high discontinuation rate. These features are probably due to low compliance in applying the natural family planning rules, which may be too demanding for a number of couples. In this review, there are comments on the application of natural family planning methods, the discontinuation rates and the failure of the method due to mistakes in the studies carried out in the past 15 years. Steps that can be taken to limit mistakes and discontinuity are also addressed. PMID- 10836193 TI - A prospective, randomized, controlled clinical study of a new subcutaneous, purified, urinary FSH preparation for controlled ovarian hyperstimulation in in vitro fertilization. AB - The aim of this study was to evaluate the clinical efficacy and safety of a new urinary follicle stimulating hormone (FSH) preparation (Fostimon) in patients undergoing in vitro fertilization-embryo transfer or intracytoplasmic sperm injection. Metrodin HP was adopted as a reference drug, as its purity and therapeutic efficacy are well known. Sixty normo-ovulatory patients aged 18-38 years with normal basal FSH and body mass index < 25 kg/m2 were selected for the study. After gonadotropin releasing hormone analogue pituitary desensitization, patients were randomized to receive either Fostimon or Metrodin HP at the initial dosage of 225 IU for 6 days. Thereafter, the dosage was tailored according to the ovarian response. Both drugs were administered by the subcutaneous route. The primary end-points were: number of follicles larger than 15 mm, levels of 17 beta estradiol on the day of human chorionic gonadotropin (hCG) injection and number of oocytes recovered. The secondary end-points were: number of FSH ampules used, day of hCG injection and pregnancy rate. FSH kinetic curves were calculated during the treatment period with both products. Safety was evaluated by pre- and post-treatment blood biochemistry and hematology, and recording all side-effects. Local tolerance was investigated at each visit. None of the parameters evaluated showed a statistically significant difference between the two groups. Local tolerance was always recorded as good/excellent by both the patients and the physician. In conclusion, Fostimon proved to be an effective and safe drug for assisted reproductive cycles. PMID- 10836195 TI - A comparison of cycle control and effect on well-being of monophasic gestodene-, triphasic gestodene- and monophasic desogestrel-containing oral contraceptives. Gestodene Study Group. AB - This was an open-label multicenter study to compare the cycle control and effect on well-being of two oral contraceptives containing gestodene and one containing desogestrel. A total of 2419 healthy women < or = 41 years of age were randomized to receive oral contraceptives containing monophasic gestodene (Minulet; n = 806, mean age 24.5 years), triphasic gestodene (Tri-Minulet; n = 808, mean age 24.6 years) or monophasic desogestrel (Mercilon; n = 805, mean age 24.6 years). Subjects were to participate in the study for up to 13 treatment cycles. A modified Moos Menstrual Distress Questionnaire was used to evaluate menstrual symptoms and to assess overall well-being. A total of 698 women were withdrawn from the study, 154 due to adverse events. Cycle control with gestodene was superior to that with desogestrel at almost all time points, particularly for breakthrough bleeding and/or spotting, which occurred significantly less frequently with gestodene than with desogestrel at cycles 1-7 and 9-11 (p < 0.05). Generally, the proportion of subjects with breakthrough bleeding and/or spotting was almost twice as great with desogestrel as with gestodene. The duration of bleeding was not consistently different between the gestodene and desogestrel groups; however, the intensity of bleeding was greater with gestodene at all time points (p < 0.05). The latent period before withdrawal bleeding was significantly longer for monophasic gestodene at cycles 1-5 and 8-10 (p < 0.05). Treatment significantly improved overall well-being at cycles 6 and 9 with triphasic gestodene and at cycle 13 with desogestrel; however, no statistically significant differences among treatment groups in overall well-being scores or individual factors of well-being could be identified. All three treatments were well tolerated. The most common drug-related adverse events were headache (14.2%), breast pain (6.2%), nausea (4.1%), metrorrhagia (3.9%) and abdominal pain (3.5%). The incidence of adverse events in all treatment groups was similar, with the exception of metrorrhagia, which occurred in more patients in the desogestrel group than in the gestodene treatment groups (p < 0.05). PMID- 10836196 TI - Demonstration of reserved anterior pituitary function among patients with amenorrhea after postpartum hemorrhage. AB - To demonstrate the residual pituitary function of patients with Sheehan's syndrome years after the obstetric complication, 14 patients with postpartum hemorrhage followed by secondary amenorrhea and agalactia were included in this review. Due to their unfamiliarity with the clinical symptoms, these patients did not receive pretreatment hormonal therapy. The mean age at their last delivery was 29 years (range 21-38 years). The mean duration between postpartum hemorrhage and the subsequent clinical manifestations leading to the endocrine investigation was 18 years (range 1-33 years). Eight patients presented with symptoms of severe hyponatremia (serum sodium less than 125 mmol/l) more than 16 years (mean 23 +/- 10) after the occurrence of postpartum hemorrhage. The electrolyte abnormality was primarily due to adrenal dysfunction. Seven out of 14 patients had normal basal luteinizing hormone (LH) levels and adequate LH responses to gonadotropin releasing hormone stimulation. Administration of thyrotropin releasing hormone provoked thyrotropin release and/or prolactin secretion in four cases. The manifestation of clinical hypopituitarism and the degree of empty sella on computed tomography scanning did not accurately indicate the secreting ability of the pituitary in patients with Sheehan's syndrome. Although all the patients had amenorrhea, the gonadotropic functions of the pituitary still remain in some patients. Various degrees of other pituitary functions can also been demonstrated even several decades after the occurrence of obstetric complications. Our data suggest that the amenorrhea of Sheehan's patients is not simply due to a dysfunction of the pituitary gonadotrophs. PMID- 10836197 TI - A case of 45,X Turner syndrome with spontaneous ovulation proven by ultrasonography. AB - A well-documented case of non-mosaic Turner syndrome, with spontaneous pubertal development and ovulatory cycles is reported. Mosaicism could be excluded both by karyotyping of 172 metaphases of blood lymphocytes and fibroblasts, and by fluorescence in situ hybridization, using an X-centromeric probe, in 200 blood lymphocyte nuclei. This Turner syndrome patient underwent normal pubertal development, with spontaneous menarche at 14 years, followed by regular monthly periods. Hormonal measurements performed during puberty were consistent with the patient's pubertal development. At the age of 26 years the patient was referred for complete fertility evaluation. Detailed hormonal analyses were performed in a given cycle. They showed midluteal phase estradiol and progesterone values within the range corresponding to normal ovulation and corpus luteum function. In the same cycle, pelvic ultrasonography was also performed at days 13, 15 and 18. It demonstrated a spontaneous ovulation, with follicular rupture that occurred between days 15 and 18. This is the first report of a spontaneous ovulation in Turner syndrome evidenced, not only by hormonal analysis, but also by ultrasonographic demonstration of follicular rupture. PMID- 10836198 TI - E- and P-selectin expression in endometriotic tissues and the corresponding endometria. AB - Endometriosis is one of the most frequent diseases in gynecology. It is histologically defined as a non-malignant pathology in which endometrial-like tissue is found outside the uterus. The pathogenesis and mechanisms involved in the development of endometriosis-associated pain symptoms are still under investigation. A local peritoneal inflammation seems to play an important role in the origin of these symptoms. Selectins belong to a group of cell adhesion molecules that mediate the localization of circulating leukocytes on the endothelium at the sites of inflammation. The aim of this study was to investigate the expression of E- and P-selectins in endometriotic tissues and the corresponding endometria. Thirty endometriotic samples, 13 corresponding endometria and six endometria of patients without endometriosis were analyzed using an immunohistochemical technique. Just two endometriotic tissues expressed E-selectin, while five samples were positive for P-selectin. The selectin expression of the corresponding endometria was also very weak. No correlations between the revised American Fertility Society (rAFS) score or the hormonal situation of the patients at the time of biopsy and the selectin expression could be found. In conclusion, the selectin expression in endometriotic glands does not play an important role in the initiation of inflammatory processes caused by endometriosis. This inflammation must be considered as a secondary reaction after the implantation of the endometriotic glands, so that endometriotic tissues are not able to induce, by the expression of selectins, a direct inflammation. PMID- 10836199 TI - Induction of estrogen receptor-alpha and -beta activities by synthetic progestins. AB - The cellular action of steroid hormones is mediated by specific receptors. Recently, two different estrogen receptors (ER), alpha and beta, have been cloned with a specific tissue distribution. Active estrogen as well as active progestin are compounds of oral hormonal contraceptives and hormone replacement therapy. To examine the regulation of ER-alpha and -beta activities after treatment with synthetic progestins and synthetic and natural estrogens, COS 7 cells were transfected with the vector expressing ER-alpha and -beta in combination with a luciferase reporter vector. ER-alpha activity was upregulated in the presence of synthetic progestins in a dose-dependent manner. Norethisterone, norethynodrel and desogestrel proved to be the most potent stimulatory agents of ER-alpha expression. On the other hand, not all progestins exhibited a stimulatory action on ER-beta activity. Only norgestrel, levonorgestrel, norethynodrel and norethisterone induced ER-beta-activating functions in a dose-dependent manner. Luciferase activity due to estrogen stimulation served as a positive control. Our results indicate that progestins have different effects on the activities of ER alpha and -beta. PMID- 10836200 TI - Hypothyroidism among infertile women in Finland. AB - The main aim of this retrospective study was to evaluate the occurrence of hypothyroidism among Finnish women with infertility. For this purpose, the records of 335 women presenting for the first time with infertility at the outpatient clinic of reproductive endocrinology at Turku University Central Hospital during a 3-year period (January 1992 to December 1994) were reviewed. Due to missing data, 36 women were excluded from the analysis. Thyroid function was screened by measuring serum thyroid stimulating hormone (TSH) levels in conjunction with serum prolactin using immunoradiometric assays. Prior to enrolment in the infertility examinations, ten out of 299 women had used thyroxine substitution for primary hypothyroidism. In the TSH screening test, 12 women (4%) exhibited elevated serum TSH levels ranging from 5.7 to 32 mU/l. Three of these cases were previously diagnosed with hypothyroidism and were using an inadequate dose of thyroxine. The prevalence of abnormal TSH levels was highest in the ovulatory dysfunction (6.3%) and unknown infertility (4.8%) groups and lowest in the tubal infertility (2.6%) and male infertility (1.5%) groups, although no statistically significant differences between the groups were observed. Oligo/amenorrhea was present in 101 (34%) women in the whole study population and in eight (67%, p < 0.5) women with elevated serum TSH at screening. The relatively high occurrence of abnormal TSH levels in infertile women with ovulatory dysfunctions or unknown infertility, as well as with oligo/amenorrhea, emphasizes the importance of TSH screening in these patient groups. PMID- 10836201 TI - The psychological outcome of hysterectomy. AB - The objective of this study is to review the published literature on psychological outcome of hysterectomy and oophorectomy for non-malignant indications. The relevant publications over the past 30 years until the end of 1997 were identified by a MEDLINE computer search. This was followed by hand searches of the relevant references in the literature identified by the electronic search. The published studies on the psychological outcome of hysterectomy have been selected to identify the incidence, possible causes and risk factors of psychological morbidity, and the measures that can be adopted to improve the outcome. The study showed that the majority of retrospective studies reported an adverse psychological outcome after hysterectomy. However, all prospective studies showed that the incidence of depressed mood is higher even before hysterectomy, owing to pre-existing psychiatric illness and personality and psychosocial problems, as a result of the emotional response to gynecological symptoms or as a manifestation of associated ovarian failure. Hence, the therapeutic effects of hysterectomy include improvement of mood in some but not all patients, unless proper case selection, psychiatric evaluation and preoperative counselling are arranged. An early detection of ovarian failure after hysterectomy, the initiation of hormone replacement therapy (HRT) immediately after surgery in perimenopausal women and in those undergoing oophorectomy, as well as regular follow-ups to ensure long-term compliance with HRT, would also improve the psychological outcome. In conclusion hysterectomy itself is not the cause of any adverse psychological outcome. Psychological symptoms actually improve in the majority of women, with the relief of distressing gynecological symptoms and the correction of ovarian hormone deficiency, but hysterectomy may not be of any benefit in women with prior psychiatric illness and those with personality and psychosocial problems. PMID- 10836202 TI - Indirect evidence that estrogen replacement therapy stimulates nitric oxide synthase in postmenopausal women. AB - The aim of the study was to investigate the effects of estrogen replacement therapy (ERT) on nitric oxide (NO) activity in healthy postmenopausal women. The study group consisted of 22 postmenopausal women (last menses at least 12 months prior to study entry) who were randomized to receive treatment for 2 months with patches that delivered either 50 micrograms/day of 17 beta-estradiol or placebo in a cross-over design. Blood samples for measurements of serum citrulline and arginine were collected at the start of the study and at the end of each treatment course. Serum citrulline and arginine were measured using high performance liquid chromatography with fluorometric detection. Arginine levels were significantly lower in the ERT group compared to the placebo group, while citrulline levels did not change. The percentage citrulline/arginine ratio was significantly higher in the ERT group (42.9 +/- 21.6) compared to the placebo group (33.9 +/- 18.5) (p < 0.01). The citrulline/arginine ratio, both at baseline and during either ERT or placebo administration demonstrated a positive linear correlation with body mass index (BMI). No correlations were found between follicle stimulating hormone, estradiol and insulin levels and BMI. No correlations were found between age, time since menopause and baseline arginine and citrulline levels or the citrulline/arginine ratio. These data indirectly demonstrate that transdermal estradiol replacement in postmenopausal women is able to stimulate NO production through the involvement of endogenous L-arginine. A positive linear correlation was found between BMI and the citrulline/arginine ratio, suggesting an additional protective cardiovascular effect in overweight women. PMID- 10836203 TI - A review of the natural occurrence, synthetic production and use of carcinogenic hydrazines and related chemicals. AB - This writing details the natural occurrence, synthetic production and use of the carcinogenic hydrazines and related chemicals. Twenty-three such chemicals were found in nature in mushrooms, tobacco, bay leaves, antibiotics, soil and other sources. The synthetically-produced hydrazines and related chemicals include 61 compounds which were or are in use as pharmaceutical drugs, agricultural substances, industrial chemicals and fuels for military and space vehicles and rockets. Because there is an overlap between the naturally-occurring and synthetically-produced compounds, their total number is 76. Altogether, 98 hydrazines and related chemicals were studied for carcinogenic action thus far, of which 84 were found to be carcinogenic and the remaining 14 were inactive. This means that the human population is exposed in various degrees to a substantial number of these agents. Therefore, the hydrazine class, unlike the polycyclic aromatic hydrocarbons, N-nitroso compounds, and aromatic amines, poses a greater environmental risk to the human population. In view of these considerations, it appears reasonable to proceed with additional experimentation in this field of interest. PMID- 10836204 TI - Effects of far-infrared ray on reproduction, growth, behaviour and some physiological parameters in mice. AB - The effects of chronic exposure to far-infrared ray (FIR) on reproduction, growth, behaviour, survival time and some related parameters were examined in SHN mice. The reproductive parameters differed slightly between the females on the normal racks and those on the FIR racks, which emitted FIR from the ceiling. The age and body weight on the day of vaginal opening was lower in the experimental mice born and maintained on the FIR rack than in the control on the normal rack. In both sexes, the levels of urinary components in the experimental group was significantly higher than the control at 6-7 months of age. Spontaneous motor activity of females during the light and dark phases were higher and lower, respectively, in the experimental group than the control. The survival rate was significantly higher in the experimental group than the control. These findings suggest that FIR has 'normalization effects' on the organisms. PMID- 10836205 TI - In vivo effect of lipoic acid on lipid peroxidation in patients with diabetic neuropathy. AB - BACKGROUND: The diabetic state, in both humans and experimental animals, is associated with oxidative stress. Lipid peroxidation of nerve membranes has been suggested as a mechanism by which peripheral nerve ischemia and hypoxia could cause neuropathy. Lipoic acid (LA) is a powerful inhibitor of iron-dependent lipid peroxidation and reactive oxygen species. The treatment of diabetic peripheral and cardiac autonomic neuropathy with LA is based on good clinical and experimental evidence. MATERIALS AND METHODS: To investigate the magnitude of the oxidative stress, serum ceruloplasmin (Cp) and lipid peroxide (Lp) levels were measured in 10 patients with diabetic neuropathy, before and 70 days after treatment with single dose of 600 mg LA/day. For other 12 healthy age- and sex matched control subjects the serum Cp and Lp levels were evaluated. RESULTS: Our results show that hyperglycemia is a factor for an increase in serum ceruloplasmin in patients with diabetic neuropathy compared to healthy subjects (p < 0.0001). High serum ceruloplasmin (Cp) level in patients with diabetes may be related to antioxidant defense. The treatment of diabetic neuropathy with LA does not affect significantly the serum Cp activity. The serum Lp levels after LA administration were significantly lower (p < 0.005) than those before treatment. CONCLUSIONS: The antioxidant therapy with LA improves and may prevent diabetic neuropathy. This improvement is associated with a reduction in the indexes of lipid peroxidation. Oxidative stress appears to be primarily due to the processes of nerve ischemia and hyperglycemia autooxidation. PMID- 10836206 TI - Interleukin-11 enhancement of VLA-5 mediated adhesion of CD34+ cells from cord blood to fibronectin is associated with the PI-3 kinase pathway. AB - Adhesion is required for cell growth, differentiation, survival, and function. Cell adhesion is mediated by a structurally diverse group of plasma membrane receptors, each exhibiting specialized ligand-binding properties that are needed for specific tasks. Integrin-mediated adhesion is important for hematopoietic stem (HSC)/progenitor (HPC) cell survival and may prevent programmed cell death. Interleukin (IL)-11, a multi-functional cytokine secreted by the bone marrow environment, plays an important role in regulating growth and differentiation of HSCs/HPCs. In this report, we demonstrate that IL-11 enhanced adhesion of freshly isolated and 3 day-expanded CD34+ cells to immobilized fibronectin. the expression of very late antigen (VLA)-4 and VLA-5 integrins was detected on CD34+ cells. CD34+ cells also expressed a-chain and gp130 subunits of the IL-11 receptor (R). Enhanced adhesion by IL-11 was mediated via activation of VLA-5 integrins, since this action could be blocked by monoclonal antibodies against beta 1 and alpha 5, but not alpha 4, integrins. Addition of phosphatidylinositol (PI)-3 kinase inhibitors blocked IL-11 enhanced adhesion of CD34+ cells to fibronectin. The results suggest that this enhanced adhesion is associated with the PI-3 kinase pathway, an inside-out signaling pathway. PMID- 10836207 TI - Intertumor linkage of age-adjusted incidence rate in 15 human neoplasias of both sexes. AB - We report here that the application of the least square method of Gauss to the log-transformed age-adjusted incidence rate changes in time and space, as tested with either the male-female or the female-male tumor pairs for each of 15 tumor entities, has revealed the presence of intertumor linkage that was conditioning the changes of two cancer risk parameters to let them fit to the equilibrium model with close resemblance to the chemical equilibrium model. The dissimilarity of the cancer risk equilibrium model to the chemical equilibrium model- topological dissociation between the equilibrium model of centripetal force (r = 1.000) and that of centrifugal force (r = +1.000)--was discussed in the light of the concept of the oncogene activation-tumor suppressor gene inactivation. The proposed network hypothesis of human neoplasia found supporting evidence in the corresponding changes of the statistical features of human neoplasias with and without sex discrimination of cancer risk. PMID- 10836208 TI - Establishment of a biological indicator for the radiation and safety of diagnostic ultrasound using apoptosis. AB - We have studied, by a nonisotopic in situ end-labeling (ISEL) technique, the frequency of apoptosis in the intestinal crypt cell of adult mice and in the external granular layer(EGL) of the cerebellum of fetuses by gamma-ray irradiation from 60Co or diagnostic ultrasound exposure. The extent of changes following 200 cGy(1090 cGy/min) was studied at 2, 4, 6, 8, 12, or 24 hours after exposure. The maximal frequency was found 4-8 hours after exposure. The mice that received 18, 36, 54, 108, 198, 396 cGy of gamma-rays or diagnostic ultrasound (7.5 MHz, 4.2 mW, ISPTA = 7.9 mW/cm2, IsppA = 114.3 W/cm2) for 10 or 30 minutes were examined 6 hours after irradiation. Measurements performed after gamma-ray irradiation showed a dose-related increase in apoptotic cells in each of the mice studied. The dose-response curves were analyzed with alpha linear-quadratic model: the frequency (number per crypt) of apoptotic cells in the intestinal crypt of adult mice was y = (0.0386 +/- 0.004204)D + (-0.0000535 +/- 0.00001120)D2 + 0.15475(r2 = 0.952, y = apoptotic cell per crypt cell, D = dose in cGy), and the frequency (percentage of apoptotic cell in the EGL) of apoptotic cell in the EGL of fetus was y = (0.1349 +/- 1.175)D + (-0.001522 +/- 0.334)D2 + 0.0477(r2 = 0.981, y = % of apoptotic cell in the EGL, D = dose in cGy). In the experiment of ultrasound exposure, the frequencies of apoptosis were 0.181 +/- 0.055(10 minutes exposure) and 0.325 +/- 0.294 (30 minutes exposure) in the crypt cells and 0.106 +/- 0.130% (10 minutes exposure) and 0.167 +/- 0.220%(30 minutes exposure) in the EGL. We estimated the relative dose of the yield from the experiment with ultrasound by substituting the yield from ultrasound exposure into the curue from the gamma-irradiation. The relative doses of ultrasound exposure compared with gamma-irradiation were 0.692 cGy(10 minutes exposure) and 1.334 cGy(30 minutes exposure) in the experiment for crypt cells and 0.432 cGy(10 minutes exposure) and 0.885 cGy(30 minutes exposure) in the experiment for EGL. Although there is presently no evidence to indicate that diagnostic ultrasound involves a significant risk, it is not wise to use diagnostic ultrasound indiscriminately. PMID- 10836209 TI - Antimicrobial and radical modulation activity of AV-07, a poly-herbal formula. AB - An extract of AV-07 was investigated for various biological activities. Pretreatment of mice with the AV-07 extract significantly protected them from lethal infection with E. coli. ESR spectroscopy showed that the extract produced radicals under alkaline conditions and enhanced the radical intensity of sodium ascorbate, suggesting its pro-oxidant action at higher concentrations. The extract effectively scavenged superoxide anion, produced by hypoxanthine-xanthine oxidase reaction and hydroxyl radical, produced by Fenton reaction. These data demonstrate that AV-07 extract contains various bioactive substances, suggesting its medicinal efficacy. PMID- 10836210 TI - Antitumour and antimetastatic effect of genistein alone or combined with cyclophosphamide in mice transplanted with various tumours depends on the route of tumour transplantation. AB - In the present study we evaluated the antitumour effect of genistein alone, cyclophosphamide alone and as a combined therapy with both agents in mice transplanted with B16F-10 melanoma and Lewis lung cancer cells. The influence of the route of inoculation of the tumour cells on the antitumour and antimetastatic effects of these therapeutics was evaluated. The antitumour effect of genistein in mice with B16F-10 intradermically (i.d.) growing tumours and in mice with LL2 subcutaneous (s.c.) tumours was observed. In addition, its antimetastatic effect (reduction of lung colonies) in mice inoculated intravenously (i.v.) with B16F-10 and in mice with LL2 cells injected either intravenously or subcutaneously was observed. No life span prolongation of mice injected intraperitoneally (i.p.) with B16F-10 cells was observed, either after treatment with genistein alone or with cyclophosphamide alone. The synergistic effect of both agents in combined treatment, when the cells of B16F-10 melanoma were injected i.p., i.v. or i.d. and in a weaker manner when the cells of LL2 cancer were injected s.c., was observed. When LL2 cells were injected intravenously, no additive effect of genistein and CY could be detected. In conclusion, we have described the experimental mouse tumour models in which both the antitumour and antimetastatic effects of genistein alone, CY alone and those of combined therapy with genistein and cyclophosphamide were dependent on the implantation route of the tumour cells. PMID- 10836211 TI - Ascorbic acid (vitamin C) effects on withdrawal syndrome of heroin abusers. AB - BACKGROUND: Ascorbic acid (vitamin C), administered orally in high doses has been observed to relieve pain and reduce opioid use in cancer patients. In vitro studies have also shown that antioxidants, such as vitamin C, may, at high concentrations, inhibit the endogenous opioid degrading metalloenzyme and increase endorphin levels. In the present study the effects of oral administration of high doses of vitamin C on withdrawal syndrome of heroin abusers were investigated. MATERIALS AND PATIENTS: Ascorbic acid at doses of 300 mg/kg b.w/day, supplemented with vitamin E (5 mg/kg b.w/day), was orally administered in two groups of heroin addict subjects consisting of in-patients (Group A, 30 males) and one of out-patients(Group B, 10 males), for a minimum of 4 weeks. The group A in-patients were also administered the conventional (diazepam + analgesic) medication. The results on the intensity of withdrawal syndrome (WS), estimated according to DMS-III criteria, were compared to a third group of heroin addict in-patients (group C, 30 males-control group), treated only by conventional medication. RESULTS: The patients of the vitamin C-treated groups (in-patients and out-patients) experienced mild WS (in 46.6% to 50% of the subjects) in contrast to the control group patients, who experienced mild WS in 6.6% of the cases. The vitamin C-treated subjects expressed major WS ranging from 10% to 16.6%, in contrast to the untreated subjects (control group), who expressed a major WS in 56.6% of the cases. CONCLUSIONS: The results indicate that high doses of ascorbic acid administered orally, may ameliorate the withdrawal syndrome of heroin addicts. Further studies are needed in order to estimate the dose- and time-dependent effects of ascorbic acid treatment, and to clarify its mechanisms of action in the withdrawal syndrome. PMID- 10836212 TI - Papillary microcarcinoma of the thyroid: a clinico-pathologic and prognostic review. AB - Papillary microcarcinoma (PMC) of the thyroid is the most common form of thyroid cancer, which usually remains clinically silent until its incidental histologic diagnosis in autopsy or surgical material. Autopsy incidence varies from 3%-36%. PMC may, however, present with clinical symptoms, the commonest of which is cervical lymphadenopathy with or without palpable thyroid nodules. Other reported presentations include cystic neck mass, pulmonary mass (es), metastases in the skull or vertebral column. The upper limit of size to define PMC is 10 mm in most studies but many studies include lesions up to 15 mm in diameter. Histologic variants include encapsulated and partially encapsulated papillary carcinoma, circumscribed microcarcinoma and occult sclerosing carcinoma. Younger age and size less than 10 mm (< 15 mm in other studies) are considered to be favorable prognostic factors. Size alone, however, cannot be regarded as a determinant of prognosis. Older age, larger tumor size, distant metastases, capsular invasion and multifocality indicate unfavorable prognosis. Loss of heterozygosity (LOH) is an infrequent finding, since small deletions may be missed by southern blot analysis. Activation of oncogenes ret and trk have been reported in papillary carcinoma. Some authors advocate conservative while others favor aggressive therapy including total thyroidectomy with or without Iodine 131ablation. Additional investigative techniques are needed to identify the subset of PMC cases with a potential for aggressive clinical course, thereby targeting more aggressive therapy to an appropriate subset of tumors. PMID- 10836213 TI - Effects of temperature strategy during cardiopulmonary bypass on cerebral oxygen balance. AB - BACKGROUND: Cerebral injury is the most serious complication of cardiopulmonary perfusion (CPB). With the advent of warm heart surgery, the effect of temperature strategy during perfusion and its effect on cerebral oxygen balance needs further study. METHODS: Three groups of patients (n=8, each) undergoing coronary artery bypass graft (CABG) surgery were studied. Group H, M and N underwent CPB at 28 degrees C, 32 degrees C and normothermia (>36 degrees C), respectively. The extracorporeal circuit was primed with Hartmann's solution 2.5 l and flow of 1.8 l min(-1) m(-2) at 28 degrees C, and 2.4 1 min-1 m(-2) at 32 degrees C and normothermia. All patients had a 4F oximetry catheter (Opticath, Oximetrix, Abbott Laboratories) inserted in the right jugular bulb for continuous measurement of jugular venous oxygen saturation (sjvO2). Data was collected at six specific times: T1- within 5 min before initiation of CPB, T2 - within the first minute after CPB, T3 - during stable temperature on CPB (28 degrees C, 32 degrees C, >36 degrees C), T4 - during rewarming at 34 degrees C in groups H and M, 15 min before coming off CPB in group N, T5 - 15 min after CPB, T6 - skin closure. At each time mean arterial pressure (MAP) and sjvO2 were recorded. Arterial blood and jugular venous blood were sampled for measurement of arterial oxygen saturation (saO2) and jugular venous lactate (sjv(lactate)), respectively. RESULTS: SjvO2 values decreased at times T2, T3 and T4 when compared to baseline (p<0.05) but there were no significant group differences at any time. Cerebral arteriovenous oxygen saturation (a-jvO2) differences mirrored sjvO2 changes. Sjv(lactate) values increased from baseline following CPB but fell consistently with time - there were no significant group differences at each time point. SjvO2 and a-jvO2 values were not significantly correlated with sjv(lactate). CONCLUSION: During pump flows employed in this study, cerebral oxygen balance and perfusion appear unaffected by temperature. PMID- 10836214 TI - Video-endoscopic dissection of multiple pedicled arterial grafts for use in minimally invasive coronary artery bypass surgery. AB - BACKGROUND: We evaluated the possibility of video-endoscopic dissection of both internal thoracic arteries (rTAs) through the left thoracic approach and right gastroepiploic artery (GEA) via a small laparoscopic access, for use in minimally invasive coronary artery bypass surgery. METHODS: The procedure was performed on twenty-two mongrel dogs. Three 10 mm ports were inserted in the left hemithorax to enable the introduction of a rigid 0-degree videoscope and forceps which allowed access to the left and right ITAs. For dissection of the GEA, one 10 mm and two 5 mm ports were inserted in the para-umbilical region. RESULTS: The left and right ITAs and the GEA were easily visualized and dissection and complete mobilization was achieved without injury, which was checked by the presence of good blood flow. The ITAs and GEA were divided distally, exteriorized through a small left anterior thoracotomy and the length of these three grafts were able to reach all of the left coronary artery branches. CONCLUSIONS: We have demonstrated, in dogs, the feasibility of video-assisted dissection of both the left and right ITAs through the left thoracic approach, without sternotomy, avoiding the risks of sternal complications and expanding its use for all patients. Furthermore, a third arterial conduit (the RGA) can be used without requiring laparotomy. PMID- 10836215 TI - Comparison of retrograde vs simultaneous ante/retrograde cold blood cardioplegia. AB - BACKGROUND: To evaluate the homeostasis of myocardium during simultaneous continuous retrograde and antegrade cardioplegia vs retrograde continuous cardioplegia. METHODS: 40 patients who underwent elective operation of coronary arteries bypass grafting were randomly assigned to 2 groups: group one consisted of 24 patients who received retrograde continuous blood cardioplegia; group two consisted of 16 patients who received simultaneous continuous ante/retrograde cardioplegia. The following measurements were taken: acidosis, oxygen content, oxygen extraction and oxygen consumption; they were taken before and after cross clamp releasing from coronary sinus effluent and from arterial line. Incidence of low cardiac output, ventricular fibrillation, raised cardiac enzymes and ischemic changes on ECG was noted. RESULTS: In simultaneous group such parameters as acidosis, oxygen content, oxygen extraction and myocardial oxygen consumption recovered after cross-clamping and changes of their values were respectively: 0.0005, 0.87 ml/100 ml, 0.098 and 1.4 ml/min. The same parameters didn't recovered in retrograde group and changes were respectively: 0.05 - p=0.2; 3.7 ml/100 ml - p=0.006, 0.29 p=0.006 and 7.4 ml/min - p=0.03. These changes were significant between groups. CONCLUSIONS: Metabolic viability of myocardium measured with oxygen utilisation is better preserved with simultaneous antegrade and retrograde cardioplegia. PMID- 10836216 TI - Complete atrioventricular septal defect associated with tetralogy of Fallot. Favourable outcome of transatrial transpulmonary repair. AB - BACKGROUND: Complete correction of atrioventricular septal defect (AVSD) associated with tetralogy of Fallot (TOF) has been reported to account for an increased surgical risk. Impaired right ventricular function after classic transventricular repair, residual outflow tract stenosis, and incompetence of the pulmonary or atrioventricular valves are considered to be essential factors affecting the results. METHODS: From 3/95 to 6/98 six consecutive patients with AVSD and TOF underwent repair (age 18 months to 7.3 years) using a combined transatrial-transpulmonary approach. RV outflow tract balloon dilatation preceded transatrial correction in 4 patients. Pulmonary annulotomy but not transanular patching was necessary in 4 cases. The septal defects were closed by two separate patches using a Dacron patch with short depth and anterior extension for the ventricular component. RESULTS: All patients survived and had stable sinus rhythm. Echocardiography demonstrated mild, but hemodynamically insignificant mitral regurgitation in two and tricuspid regurgitation in four patients. Right ventricle to pulmonary artery gradients ranged from 5 to 35 mmHg (mean 24.2 mmHg) without progression. During follow-up ranging from 4 months to 3.5 years (mean 16.8 months) no reoperation was necessary. CONCLUSIONS: The transatrial transpulmonary approach for correction of AVSD with TOF contributes to improved results after repair of this rare combination of defects. PMID- 10836217 TI - Outcome of patients with cyanotic congenital heart disease undergoing a second systemic-to-pulmonary artery shunt. AB - BACKGROUND: Multiple systemic-to-pulmonary artery shunts often represent a deviation from the initial management strategy for cyanotic congenital heart disease. This study analysed the outcome in patients undergoing a second shunt. METHODS: Between 1965 and 1998, 80 patients required a second shunt. Patient age ranged from 11 days to 11.2 years at the initial shunt, and 4 months to 25.6 years at the second shunt. The interval between shunts ranged from 11 days to 19.6 years. RESULTS: The patient survival rates at 1, 6, 12, 60 and 120 months following the second shunt were 95.0 +/- 2.4%, 92.5+/-3.0%, 87.4+/-3.7%. 82.4+/ 4.5% and 74.8+/-5.8%, respectively. Although excess pulmonary flow was not encountered after the second shunt, mode and incidence of morbidity were otherwise similar to the first one. Between 9 months and 7.5 years after the second shunt, 24 patients underwent successful definitive repair. Multivariable analysis identified pulmonary atresia (p=0.027) and a short (<1 year) interval between the two shunts (p=0.034) as the independent risk factors for long-term shunt failure. Single ventricle physiology (p=0.002) and a central approach for the second shunt (p=0.016) were independent risk factors for lack of application, or failure of intracardiac definitive repair. CONCLUSIONS: A significant limitation in longevity and quality of life is common in patients requiring a second shunt, especially those associated with pulmonary atresia, the single ventricle physiology heart, or ungraftably hypoplastic contralateral branch pulmonary artery to the first shunt. Since excess pulmonary blood flow leading to congenital heart failure and/or ventricular diastolic dysfunction is unlikely, all patients who preclude definitive repair due to decreased pulmonary blood flow even after the first shunt should be shunted again. PMID- 10836219 TI - A mathematical analysis of hemorheological changes during heart valve replacement. AB - BACKGROUND: This study was carried out to establish a mathematical model in order to assess blood trauma and hemorheology during cardiopulmonary bypass and heart valve replacement. METHODS: Ten factors which represented blood trauma and hemorheology were investigated in fourteen patients undergoing mechanical heart valve replacement. RESULTS: The results confirmed that red blood cell damage was mainly dependent on cardiopulmonary bypass time and hematocrit level. Platelet aggregation was influenced by platelet count, plasma fibrinogen and cardiopulmonary bypass time, the cases with aortic valve replacement resulting in more platelet activation than the mitral valve replacement (p<0.05). High shear blood viscosity was significantly influenced by hematocrit, plasma viscosity and red cell filterability, while low shear blood viscosity was significantly related to hematocrit, plasma viscosity and fibrinogen concentration, which represented 68.5% and 74.8% of hemorheologic changes due to blood trauma respectively. CONCLUSIONS: The relationship between blood trauma and hemorheologic changes was evaluated and the potential areas for improvements in cardiopulmonary bypass techniques in relation to mechanical heart valve implantation were identified. These areas of technical and pharmacological development must reduce changes in all the possible plasma components especially fibrinogen and also preserve platelets and red cells from damage. PMID- 10836218 TI - Comparison of 21-23 size St. Jude medical valves to the 27-29 size mechanical valves in aortic position. Ten-year follow-up. AB - BACKGROUND: Between 1986 and 1996, 194 patients underwent isolated aortic valve replacement with 21-23 no. St. Jude Medical mechanical heart valves (small sized group) and 163 patients with 27-29 no. (large sized group). METHODS: The mean age at operation was 45.04+/-15.90 years (range: 12-76 years) for the small sized group and 38.05+/-13.41 years (range: 16-68 years) for the large sized group. Preoperatively, 39.7% of the patients from the small sized group and 42.9% from the large sized group had pure aortic stenosis, 31.9% and 27.6% had pure aortic insufficiency. Most of the patients had rheumatic valve disease. RESULTS: The overall hospital mortality rate was 12.4% and 3.07% respectively in the small sized and large sized groups (p<0.001). The overall actuarial survival rate for 10 years was 95.33+/-2.73% and 93.06+/-3.98% respectively in the small sized group and large sized group (p>0.05). In the small sized group male sex and all complications, in large sized group age and all complications were the statistically important hospital mortality predictors (p<0.05). CONCLUSIONS: Although, operative mortality and long term morbidity were higher in the small sized group, these changes did not reflect the actuarial survivals between the groups. Small sized valves carry some risk, but these risks do not affect long term survival. PMID- 10836220 TI - Myocardial preservation in acute coronary artery occlusion with coronary sinus retroperfusion and carnitine. AB - BACKGROUND: Effectiveness of retrograde coronary sinus perfusion with the use of carnitine supplementation over the severity of ischemia/reperfusion injury, in acute coronary occlusion. METHODS: Eighteen mongrel dogs, divided equally into control, retrograde perfusion (retroperfusion) and carnitine retroperfusion (retrocarnitine) groups. After taking the basal values, the left anterior descending artery was occluded. At the fifteenth minute, without ending the occlusion, retrograde coronary sinus cardioplegia in the retroinfusion group and in the carnitine group, 0.15 mmol/kg of L-carnitine retroperfusion was performed. Then, hemodynamic and biochemical measurements were taken till the end of 120 minutes. The control group had no retroperfusion or medical therapy. RESULTS: Between the three groups, there was a statistically significant difference in cardiac index, mean arterial pressure, mean pulmonary artery pressure, pulmonary capillary wedge pressure, right atrial pressure as hemodynamic parameters and myocardial oxygen extraction, myocardial Lactate extraction, protein thiols and Malonyl dialdehyde (MBA) as biochemical parameters, at different time intervals (p<0.05). CONCLUSION: Coronary sinus retroperfusion with carnitine is found to be very effective in reducing oxygen free radical release and however this advantage did not switch to the hemodynamic function between the retrograde coronary sinus infusion group and retroinfusion carnitine group. In our opinion retrograde coronary sinus perfusion with the use of carnitine supplementation reduces the severity of ischemia/reperfusion injury. PMID- 10836221 TI - Repair of leaking postinfarction ventricular septal defect under circulatory arrest via left thoracotomy. AB - We treated a 60-year-old woman for postinfarction ventricular septal defect (VSD) and closed it by the infarction exclusion method. Postoperatively she was complicated by Candida sternal mediastinitis and residual shunt of VSD. After her sternal infection came under control we repaired the leaking VSD via left thoracotomy under hypothermic circulatory arrest. She recovered well and repair of the leaking VSD under circulatory arrest via left thoracotomy seemed to be a safe and promising alternative for VSD repair. PMID- 10836222 TI - Mitral valve repair and septal myectomy for hypertrophic obstructive cardiomyopathy. AB - Combined mitral valve repair using the sliding leaflet technique and septal myectomy were employed to successfully treat left ventricular outflow tract (LVOT) obstruction and mitral regurgitation due to hypertrophic obstructive cardiomyopathy (HOCM). A 46-year-old man was diagnosed with HOCM along with congestive heart failure and was treated medically. These symptoms, however, were resistant to medical treatments with a beta-blocker, a Ca-antagonist, and disopyramide, and he was referred to our hospital for surgery. Doppler echocardiography demonstrated an LVOT obstruction at rest with a peak pressure gradient of 138 mmHg. The interventricular septum thickness was 14 mm. Mitral regurgitation of 3+ with severe SAM was also observed. Temporary dual chamber pacing was tried without significant improvement. Following these examinations, the patient underwent surgery. A transaortic septal myotomy-myectomy was performed first, and the mitral valve was then approached through the left atrium. Mitral valve repair was performed with the sliding leaflet technique to reduce the height of the posterior leaflet from 2 cm to 1 cm. Postpump transesophageal echocardiography revealed no MR and a peak LVOT gradient of 15 mmHg. The patient recovered well except for a residual mild SAM, and MR2+. We therefore concluded that this surgical approach might provide results which are superior to those of myectomy alone. PMID- 10836223 TI - Dissection in a right sided porcine-valved Dacron conduit. AB - Seventeen years after corrective surgery for a double outlet right ventricle and pulmonary stenosis, a 30-year-old patient was admitted for reoperation for a critical stenosis of a porcine-valved Dacron conduit between the right ventricle and pulmonary artery. A residual ventricular septal defect was present. Induction of anaesthesia resulted in an inadequate pulmonary flow and uncontrollable ventricular arrhythmias. At autopsy the conduit showed a dissection between the conduit wall and the peel formation, leaving a residual lumen with a diameter of less than a third of the original. Patients with a right sided bioprosthetic valved conduit must be evaluated regularly and operated before the stenosis becomes critical. PMID- 10836224 TI - Successful conservative surgery of acute ascending aortic dissection occurring during coronary angiography. AB - We reported the case of an acute aortic dissection complicating right guiding catheter manipulation during engagement in the right coronary ostium. Despite absence of hemodynamic deterioration, dissection progressed rapidly from the sinus of Valsalva to the ascending aorta along its entire length. At surgery, performed in emergency, the aorta was not dilated and the aortic wall did not appear pathologic. Therefore conservative surgery was performed, consisting of suture of the aortic tear and incollage of the false lumen, with good immediate and mid-term results. PMID- 10836225 TI - Interposition of hydroxyapatite ceramic stents alternative to delayed sternal closure. AB - Ascending and hemiarch replacement surgery for an acute Stanford type A dissection in association with a previous type B dissecting aneurysm was performed on a 58-year-old female patient. However, sternal closure could not be performed after surgery due to hemodynamic deterioration. Even four weeks after the operation, sternal closure was impossible due to advanced adhesion around the mediastinum caused by mediastinitis. Therefore, hydroxyapatite ceramic spacers, which have osteogenesis and ossification characteristics, were interposed between the split sternum as stents to avoid further surgery. PMID- 10836226 TI - Video assisted thoracoscopic ligation of patent ductus arteriosus. Technique of sliding loop ligation. AB - Variant video-assisted thoracoscopic surgical technique for closure of patent ductus arteriosus has been introduced. Although the endoscopic clipping device may be a preferred method for interruption of the patent ductus arteriosus, there is always the risk of inadequate clip placement and limitation of application in width of ductus arteriosus. In an effort to overcome this problem we have used a self-made endoscopic loop ligation in 10 patients and herein report the method. Only a small window and one port of access are necessary to dissect the patent ductus arteriosus from the surrounding tissues and to apply the extracorporeally created sliding loop. Successful ligation without shunt is obtained in all cases. The technique is simple and safe even in the presence of a wide ductus. PMID- 10836227 TI - Quantification of carotid stenoses using 3D morphometer, CT angiography and conventional angiography. AB - BACKGROUND: The aim of this study is to compare the performances of 3D morphometer (M3D) regarding the quantification of extracranial carotid stenoses with selective 2D conventional angiography (reference technique) and spiral CT scan. METHODS: It is a prospective study, including 15 patients (mean age 75) presenting a symptomatic carotid lesion detected via duplex Doppler to be operated. Patients had to hold their breath for 20 seconds. Twenty-nine carotid bifurcations were studied by means of M3D and 2D conventional angiography (15 patients). Only 10 patients (19 bifurcations) underwent a CT scan. The measures were performed on a visual display unit by measuring the pixels as per the NASCET technique. Diameters (MIP technique) and surfaces (reformated axial slices) were measured. RESULTS: With MIP technique, a good correlation was found 20 times in 29 (69%) (overestimation: n=8/29) between M3D and angiography, but only in 9 of 19 cases (47%) between CT scan and angiography. Unlike with M3D, the slices re oriented with CT scan gave better correlations: 15 times in 19 (79%). With the CT scan, the measures were impossible 8 times on MIP technique, and twice on reformated slices for calcified tight stenoses. Therefore, the quantification by means of spiral CT scan is easier by planimetry on slices. CONCLUSIONS: The reliability of these 3D explorations regarding quantification remains uncertain. The use of a morphometer, provided that technical adaptations are made, could replace selective sequences in multiple incidences that become accessible in the post-treatment period, with an acquisition field larger than the one of the CT scan. PMID- 10836228 TI - Value of the profundapopliteal collateral index for selecting between an in-flow and sequential arterial reconstruction in patients with multisegment arterial occlusive disease. AB - BACKGROUND: It remains difficult for surgeons to choose between an in-flow and sequential arterial reconstruction in patients with multisegment arterial occlusive disease. In addition, the exact criterion for the proper revascularization procedures of these patients also remains obscure. METHODS: The profundapopliteal collateral index (PPCI) was determined in all patients with occlusions of both the aortoiliac and superficial femoral arteries prior to undergoing an arterial bypass. The PPCI in the inflow bypass (IB) was also compared with the sequential bypass (SB). RESULTS: The symptoms of all patients undergoing either IB or SB improved. Preoperatively, the average PPCI in IB patients was significantly lower than that in SB patients. In addition, no significant difference was observed in the increased average rate of the ankle brachial index (ABI) between IB and SB. CONCLUSIONS: The PPCI is an accurate predictor of the hemodynamic potential of the geniculate collaterals. In cases with a low PPCI, especially in patients with multisegment arterial occlusive disease, in-flow procedures alone may often be sufficient for the successful treatment of such patients. The PPCI is thus considered to be useful for selecting the optimal revascularization procedures. PMID- 10836229 TI - Predictive value of transcutaneous oximetry for selection of the amputation level. AB - BACKGROUND: Transcutaneous oximetry was studied to select the correct amputation level for limb ischemic necrosis with possible identification of threshold value. METHODS: The method was evaluated in 30 cases (20 patients, 5 males and 15 females, of ages ranging from 61 to 93, average 73.1+/-8.5) where patients underwent amputation because of severe leg ischemia. Surgical operations were: minor amputation (toe or transmetatarsal) in 23 cases, below knee amputation in 7. Oxygen tension was measured at the dorsum foot and at the third superior of the anteromedial calf aspect. RESULTS: Results were classified as success (primary or delayed healing) or failure (necrosis at the surgical wound). Amputation was successful in 17/30 cases with oxygen tension of 0-65 mmHg (mean 32.5+/-16.1) and failed in 13/30 with oxygen tension of 0-57 mmHg (mean 7.8+/ 17.3). The difference was statistically significant (p=0.0004). Sensitivity of the method resulted 88.2%, specificity 84.6%, diagnostic accuracy 86.7%, positive predictive value 88.2% and negative predictive value 84.6%. A threshold of 20 mmHg was identified: 15/17 successful cases showed values greater than 20 mmHg, while 11/13 failed cases presented values lower than the threshold. CONCLUSIONS: Following our observations and according to some reported studies, we believe transcutaneous oximetry to be the best method for selection of amputation level This is a simple, noninvasive and accurate method, which has showed itself superior to other techniques (i.e., Doppler and radioisotope). PMID- 10836230 TI - Iloprost protects the spinal cord during aortic cross-clamping in a canine model. AB - BACKGROUND: Surgical procedures on the thoracoabdominal part of the aorta make the spinal cord vulnerable to ischemia. Paraplegia is the most severe complication following thoracoabdominal operations. In this study, iloprost was used as an agent to decrease the severity of ischemia and reperfusion injury to the spinal cord during aortic occlusion and declamping. METHODS: Twelve adult mongrel dogs weighing 17+/-2 kg were used in this study. The animals were randomly assigned to either group I, which received saline solution (6 dogs), or group II, which received prostacyclin. Group I was referred to as the control group and group II as the iloprost group. After baseline measurements were completed, the aorta was cross-clamped for sixty minutes distal to the left subclavian artery. No pharmacologic agents were used to control blood pressure in group I. Proximal and distal mean arterial pressures (DMAP) were monitored continuously. DMAP were considered as diastolic pressure in preocclusion and reperfusion periods. Iloprost administration was started at a rate of 5 ng/kg/minute five minutes before the aortic occlusion. This dosage was increased to 25 ng/kg/minute during aortic occlusion. RESULTS: Mean proximal arterial pressure was 147+/-12 mmHg in the control group and 116+/-13 mmHg in the iloprost group at occlusion (p<0.01). Mean distal arterial pressure was 19+/-7 in the control group and 37+/-5 in the iloprost group during clamping (p<0.05). Functional outcome was evaluated according to Tarlov scores 24 hours after the study. Although none of the animals recovered completely from the control group, 4 animals from the iloprost group recovered (p<0.05). Following the neurologic assessment, animals were sacrificed and specimens were taken for the electron microscopic study. Electron microscopic changes documented that severe mitochondrial damage and vacuolisation occurred in the control group. However these changes were more subtle in the iloprost group. CONCLUSIONS: As a result of this study we concluded that iloprost infused before and during clamping of the thoracic aorta mitigates the spinal cord injury due to ischemia and reperfusion following unclamping. PMID- 10836231 TI - Rupture of infected pseudoaneurysms in patients with implantable ports for intra arterial infusion chemotherapy. AB - Intra-arterial hepatic chemotherapy via implantable reservoirs is being used increasingly. In our department, five patients have undergone emergency surgery since 1991 because of rupture of an infected pseudo-aneurysm at the site of entry of the catheter. Surgical procedures included removal of the catheter and the reservoir, and closure of the affected artery with or without reconstruction. Of these patients, three (60%) died from uncontrollable sepsis. The poor prognosis emphasizes the need, in patients with carcinoma, for strict aseptic technique and hemostasis at the time of catheter placement, and for careful device maintenance. PMID- 10836232 TI - Vagal paraganglioma. Report of a case surgically treated and review of the literature. AB - The authors report the observation of one case of vagal paraganglioma occurred in a young woman. The tumor manifested itself as a left sub-mandibular tumescence; the very first diagnostic approach was achieved through echography, which showed a mass behind the internal carotid artery and compressing the internal jugular vein. The following examinations, represented by neck CT, NMR, angiography and fine needle aspiration initially directed towards the suspicion of chemodectoma. Only at operation, the anatomical situation of the tumor, which encapsulated the vagus nerve and the subsequent results of the hystological examination revealed the correct diagnosis of vagal paraganglioma. PMID- 10836233 TI - Acute ischemia of a limb as a complication of multiple hereditary exostoses. Case report and literature review. AB - The authors present a case of acute ischemia of a limb as a complication of multiple hereditary exostoses. They discuss surgery, complications, and review the literature. PMID- 10836234 TI - Combined long-segment angioplasty and stenting of the superficial femoral artery and popliteal-distal bypass for limb salvage. AB - The treatment of multisegment superficial femoral artery and tibial vessels atherosclerotic disease is traditionally a femoro-distal bypass using in situ or reversed autogenous vein. The improved result of balloon angioplasty and stenting of the superficial femoral artery (SFA) has extended its application to treat long segment SFA stenosis. A combined endovascular-open surgery approach of SFA endovascular stenting with a popliteal-distal bypass is an alternative less invasive procedure to achieve optimum distal revascularization for limb salvage. PMID- 10836235 TI - Primary aortoduodenal fistula complicated by abdominal aortic aneurysm. AB - A 74-year-old male patient was operated in Vakif Gureba Hospital for aortoduodenal fistula developing from abdominal aortic aneurysm. The patient was diagnosed as abdominal aortic aneurysm after physical examination and computed tomography in another center. Appearing of melena and hematemesis gastroduodenoscopy and radionuclide scanning was performed as diagnosis. After 6 days gastrointestinal bleeding recurred in massive haemorrhage and the patient was operated with a diagnosis of aortoenteric fistula as emergency. A midline laparotomy was performed. There was a fistula between infrarenal abdominal aortic aneurysm (with diameter 8x10 cm) and the 3rd portion of the duodenum. The duodenum was resected segmental and the fistula was disconnected. Following aneurysmotomy a prosthetic graft was placed in the aortobiiliac position. The patient was discharged at the 42nd postoperative day. Primary aortoenteric fistula is a very rare consequence of untreated abdominal aortic aneurysm. The segments of intestine most frequently involved in aortoenteric fistula are the 3rd and 4th portions of the duodenum. Clinical presentation is recurrent episodes of gross gastrointestinal haemorrhage. These cases have high mortality and morbidity unless evaluated as quickly as possible and appropriate surgical intervention performed. PMID- 10836236 TI - Radical aortic replacement employing simultaneous modified Bentall and elephant trunk procedure in Takayasu's arteritis. AB - A case of Takayasu's arteritis resulting in extensive fusiform aneurysmal dilatation of the entire aorta extending from the aortic root to the abdominal bifurcation associated with aortic regurgitation is described. She underwent successful radical replacement of the aortic root, ascending, transverse arch and proximal part of the descending aorta employing simultaneous modified Bentall and Elephant trunk techniques. During aortic arch replacement the brain was protected by selective antegrade innominate perfusion under moderate hypothermia. A pattern to the best of our knowledge, has not been reported earlier. PMID- 10836237 TI - Preservation of renal function in reimplantation of renal artery of abdominal aortic aneurysm. AB - Intraoperative administration of diuretics and renal hypothermia with cold (4 degrees C) heparinized Ringer's lactate were useful methods for preserving renal function during warm ischemia time. 54-year and 74-year-old men were diagnosed as abdominal aortic aneurysm. Their left renal and accessory left renal arteries originated from the border zone of the aneurysm. We reported two cases of reimplantation of the renal artery in abdominal aortic aneurysm without deterioration of renal function. PMID- 10836238 TI - Cardiorespiratory function is significantly improved following corrective surgery for severe pectus excavatum. Proposed treatment guidelines. AB - BACKGROUND: Pectus excavatum is a congenital chest abnormality which may become more marked during childhood and teenage growth. Young teenagers with severe PE deformities often become short of breath during strenuous exercise and complain of becoming easily fatigued. The pathophysiologic effects of PE remain controversial because they have been difficult to measure, especially in young children. Symptomatic patients often have significant subjective improvement during exercise following PE repair and there is a major cosmetic impact on teenagers with severe deformities. This study was designed to measure pulmonary and cardiac function with exercise before and after corrective surgery. METHODS: Pulmonary function testing and incremental exercise testing were performed in 36 adolescents with pectus excavatum (PE) and 10 age-matched, healthy control subjects. Six months after corrective surgery was performed, 15 of these PE patients and 6 control subjects were re-evaluated for pulmonary function. RESULTS: Before surgery, PE subjects had a lower forced vital capacity (FVC) than controls; there was no change in FVC after surgery. Before surgery, 58% of PE patients had subjective complaints of exercise limitation; 66% of the patients were significantly improved after surgery. PE subjects exercised at a similar workload to controls. Respiratory parameters during exercise were similar between the two groups before surgery, indicating that exercise was not limited by restrictive lung disease. After surgery, PE subjects exercised longer and had a higher oxygen pulse than before surgery, whereas controls showed no such changes. Although some PE subjects showed mild restrictive lung function, surgical repair did not influence this mild degree of restriction. CONCLUSIONS: After corrective surgery, PE patients have increased exercise tolerance and a higher oxygen pulse. Oxygen pulse is a measure of cardiac output. Results suggest that PE repair improves cardiopulmonary function during vigorous exercise. Based upon these studies, and our experience in the treatment of more than 700 surgical patients with pectus excavatum over a 40-year period, guidelines for the diagnosis and management of children with pectus excavatum are proposed. PMID- 10836239 TI - Atypical bronchial carcinoids. Review of 46 patients. AB - BACKGROUND: The purpose of this study was to assess the behaviour of atypical carcinoids operated at our Department in the period 1977-1998 and to review the last 19 cases according to Capella's classification (1994), indicating the most adequate surgical approach. METHODS: On the basis of anatomo-pathological characteristics, we have reviewed surgical treatment and outcome in 46 patients, submitted in the last 22 years to surgical resection for neuroendocrine neoplasms. RESULTS: 5-year survival is 77.2%; 10-year survival is 53.2%. Lymph node metastases are also important for survival, but less than the histotype. The review of our last six years' series, according to Capella's classification, of 19 patients affected by so-called atypical carcinoids revealed that: 5 were well differentiated neuroendocrine tumors (WDNT), 12 were well differentiated neuroendocrine carcinomas (WDNC), 2 were small cell neuroendocrine carcinomas (SCLC). The 5-year overall survival of our cases is 78%, for the WDNT 100%, for WDNC 81.2%. Of the 2 patients with SCLC, one survived 2 months; the other is still alive 5 months after surgery. CONCLUSIONS: The authors conclude that 5-year and 10-year survival are strongly related to the histological type of neuroendocrine neoplasm and to the presence of lymph node metastases. Capella's anatomo-pathological classification helps to give a more accurate prognosis for survival in so-called "atypical carcinoids". If the neoplasm is malignant, the authors recommend radical resection, if possible. PMID- 10836240 TI - Subcarinal foregut cysts. A unique clinical problem. AB - BACKGROUND: To evaluate the risks associated with a subcarinal foregut cyst in a fixed mediastinal space. METHODS: DESIGN: Between January 1, 1986, and August 1, 1997, 8 patients who had subcarinal cysts and who underwent surgical intervention were identified. These results were analyzed to identify associated symptoms and results of surgical intervention. Mean duration of follow-up was 37.3+/-2.2 months and was 100% complete. PATIENTS: Of the eight patients, three were men. Mean age was 45.6+/-15.6 years (range 24-66). All patients were symptomatic. Six patient suffered respiratory distress. Four patients complained of chest pain. Preoperatively, all patients underwent routine chest radiography. Six patients underwent computed chest tomography (CT); 4 patients had magnetic resonance imagery (MRI) of the chest. Cardiac echocardiography was performed on 4 patients and esophagogastroduodenoscopy (EGD) with or without esophageal ultrasound (EUS) was done in 4. Of 7 patients who underwent bronchoscopy, 6 patients demonstrated extrinsic airway compression. The remaining patient showed fistulous communication. INTERVENTION: Cyst dimensions ranged from 7.33+/-1 cm (mean+/-SD). Total resection of the cyst was accomplished in 6 patients. One patient with fistulization underwent right main bronchial sleeve resection. Histopathology revealed inflammation of the cyst in 2 cases and calcification of the cyst wall in 1. Fluid from one cyst grew gram positive cocci. RESULTS: Length of hospital stay ranged from 10.9+/-4.4 days. There were no mortalities. Morbidity included prolonged ventilation (1), pulmonary embolism (1) and left recurrent laryngeal palsy (1). CONCLUSIONS: Our results corroborate the need for surgical excision of subcarinal subtype cysts to prevent the development of mediastinal compression and other cyst-related complications. PMID- 10836241 TI - Chronic expanding hematoma in the chest. AB - We report the successful surgical treatment of chronic expanding hematoma in the chest. Four patients who had previously undergone artificial pneumothorax, thoracoplasty or tumor extirpation more than 30 years earlier recently became aware of a slowly growing mass. Chronic expanding hematoma which developed into very large masses over a long period of time were thus successfully resected. These patients are now all in good health with no recurrence after the operation. It is important to monitor such patients' laboratory data for hemostasis including the platelet cell counts, the % prothrombin time and the D-dimer, both before and immediately after operation, and the intraoperative bleeding volume. PMID- 10836242 TI - Gastropleural fistula as complication of postpneumonectomy empyema. AB - A 54-year-old woman underwent a left pneumonectomy for monolateral congenital pulmonary cysts, complicated by a pleural empyema without bronchial fistula, in the late postoperative period. The pleural empyema was evacuated and managed by means of a small thoracic drainage. Three months after discharge the patient noticed the presence of ingesta in the pleural washing fluid. Diagnostic and operative procedures in this rare case of non malignant, non traumatic gastropleural fistula are described. PMID- 10836243 TI - Rounded atelectasis in a patient with history of asbestos exposure. A case report. AB - We report a case of a 68-year-old patient with a history of chronic asbestos exposure and a lung tumour, highly suspicious for bronchial carcinoma. The patient underwent left lower lobectomy and histology showed the rare diagnosis of rounded atelectasis. Rounded atelectasis is an important differential diagnosis to bronchial carcinoma. PMID- 10836244 TI - The hospital volume-outcome relationship in general thoracic surgery. Is the surgeon the critical determinant? AB - BACKGROUND: Complex operations, such as those performed in thoracic surgery, have a hospital volume-outcome relationship. It is difficult to isolate the effect of the surgeon in this relationship since experienced thoracic surgeons tend to practice in high-volume tertiary care hospitals. An American comprehensive cancer hospital created a community outreach satellite program in thoracic surgery, and this provided a unique opportunity to study the hospital volume-outcome relationship without the confounding variable of surgeon experience. METHODS: A retrospective review of thoracic surgical operations done over a 4-year period at a small community hospital, by a tertiary care hospital surgeon, was conducted. Operative mortality was the major outcome measure. Two high complexity operations (pneumonectomy and esophagectomy) were specifically scrutinized. RESULTS: 486 thoracic surgical procedures (317 minor and 169 major cases) were done. There was one in-hospital death (aspiration pneumonia after esophageal stenting) and one 30 day mortality (readmission for cerebral vascular accident after lobectomy). Data,for the 10 esophagectomy patients is as follows: age - 66+/-13 years; length of stay - 12.8+/-3.4 days; anastomotic leaks - 0; operative mortality - 0. Data for the 6 pneumonectomy patients is as follows: age - 69+/-8 years; length of stay - 8.5+/-5.2 days; preoperative FEV1 - 1.6+/-0.3 litres; fistulas or empyema 0; operative mortality - 0. CONCLUSIONS: Despite having a very low volume of thoracic surgical cases the community hospital had crude outcomes comparable to those reported from high volume tertiary hospitals. This suggests that the surgeon may be a more important factor in the hospital volume-outcome relationship than previously thought. Nevertheless, complex thoracic surgical operations are ideally performed by an experienced surgeon, and in a high volume hospital PMID- 10836245 TI - A femoral false aneurysm secondary to artificial leg trauma. PMID- 10836246 TI - Evidence for superantigen involvement in preeclampsia. AB - PROBLEM: Preeclampsia is the leading cause of maternal morbidity and premature fetal delivery in the United States, most likely involving the immune system in disease genesis. In this report, we tested the hypothesis that a superantigen phenomenon is an important factor in the pathogenesis of the disease. METHOD OF STUDY: A semi-quantitative polymerase chain reaction (PCR) was used to assess T cell receptor (TCR) beta chain variable (Vbeta) regions as an indicator of T-cell expansion in both peripheral blood and basal plate of preeclamptic patients. All the subjects were also molecularly typed to identify their HLA-class II alleles. RESULTS: In peripheral blood of the majority of the patients, there was a high abundance of the Vbeta4 gene family, which was not observed in the control group. Polyclonality of this Vbeta gene family was confirmed by analysis of the Valpha chain and the complementary determining region 3 (CDR3). The majority of patients carried the Human Leukocyte Antigens (HLA)-DRB1*13 allele. CONCLUSION: We present evidence for the existence of a superantigen-like effect in at least a subset of patients with preeclampsia. PMID- 10836247 TI - Fas antigen expression on the decidual lymphocytes of pre-eclamptic patients. AB - PROBLEM: Apoptosis has been proposed as a mechanism for maintaining the homeostasis in the immune system. Activated lymphocytes are removed by a programmed cell death process Fas/FasL-mediated called activation induced cell death. The aim of the study was to investigate Fas antigen expression on decidual cells (T CD4+ lymphocytes, T CD8+ lymphocytes and Natural Killer (NK) cells) of pre-eclamptic patients and healthy pregnant women. METHOD OF STUDY: 12 pre eclamptic patients and 10 healthy pregnant women were studied. Lymphocytes were isolated from decidual tissues mechanically, labeled by direct staining with monoclonal antibodies, and analyzed using the flow cytometric method. RESULTS: We found Fas antigen expression on decidual NK cells and T lymphocytes. CD 95 molecule expression and fluorescence intensity on NK cells of pre-eclamptic patients were lower when compared with controls (P < 0.05). CONCLUSIONS: These findings suggest that decidual NK cells and T lymphocytes are able to undergo Fas/FasL-mediated apoptosis. It seems that NK cells' ability to undergo Fas/FasL mediated apoptosis in pre-eclamptic patients can be altered because of lower CD95 molecule expression. PMID- 10836248 TI - Thyroid immunity, thyroid dysfunction, and the risk of miscarriage: a propos article by Vaquero et al. Mild thyroid abnormalities and recurrent spontaneous abortion: diagnostic and therapeutical approach. PMID- 10836249 TI - Mild thyroid abnormalities and recurrent spontaneous abortion: diagnostic and therapeutical approach. AB - PROBLEM: The aim of this study is to evaluate the role of mild thyroid abnormalities in recurrent spontaneous abortion, and to assess the effects of two different therapeutical protocols. METHOD: A prospective study in the population of recurrent aborters with mild thyroid abnormalities, evaluating the obstetric outcome in 42 patients. Sixteen thyroid autoantibodies positive patients were treated with thyroid replacement therapy, while 11 patients received intravenous immunoglobulins (IVIG). Fifteen patients, characterized by negative antithyroid antibodies, and having underlying thyroid pathology, were treated with thyroid replacement therapy. RESULTS: Among patients with thyroid antibodies, 6 out of the 11 pregnancies (54.5%) treated with IVIG ended in live birth. In the thyroid supplementation group, 13 out of 16 pregnancies (81.2%) ended in live birth. Only one pregnancy loss occurred among patients with a mild underlying thyroid pathology treated with thyroid replacement therapy. CONCLUSIONS: Mild thyroid abnormalities are associated with an increased rate of miscarriage. This poor obstetrical prognosis seems to be related to an impaired thyroid adaptation to pregnancy. Thyroid replacement therapy appears to be more effective than IVIG in preventing a new miscarriage. PMID- 10836250 TI - Menstrual cycle dependent fluctuations in NK and T-lymphocyte subsets from non pregnant human endometrium. AB - PROBLEM: To establish the relative proportions of endometrial lymphocyte subpopulations during the menstrual cycle. METHOD OF STUDY: Lymphocytes were investigated by flow cytometry, during the early proliferative (EP), late proliferative (LP), early secretory (ES) and late secretory (LS) phases of the menstrual cycle. RESULTS: Between the LP and LS phases, there was an increase in NK cells from 26.4% to 83.2% (P = 0.0017) of the CD45+ population. T cells decreased from 55.06% to 6.7% (P = 0.0017). Within the T-cell population, CD8+ cells decreased significantly from 63.0% in the LP phase to 54.2% in the LS phase (P = 0.04). In contrast, potential regulatory populations such as double negative (DN) T cells CD3+CD4-CD8- and natural T cells (NT) CD3+CD56+, increased significantly in the LS phase (P = 0.05; P = 0.03). gammadelta T cells, predominantly of the DN and NT phenotypes remained consistently low at all stages of the cycle (2.34%). CONCLUSION: Endometrial lymphocyte fluctuations during the menstrual cycle may reflect hormonal regulation of maternal immunity, thereby promoting tolerance at the time of implantation. PMID- 10836251 TI - Detection of chlamydial antigenic material in ovarian, prostatic, ectopic pregnancy and semen samples of culture-negative subjects. AB - PROBLEM: The pathogenesis of long-term sequelae in Chlamydia trachomatis infection is poorly understood. While serology indicates previous chlamydial infection, culture studies are frequently negative. We wanted to know whether in chronic cases the bacterium is absent or persists in a dormant state where it evades detection. METHODS OF STUDY: Using immunoperoxidase (IP) staining and in situ hybridization (ISH), we examined tissues of culture-negative subjects. Ovarian biopsy specimens from 19 culture-negative women with pelvic adhesions and/or tubal infertility were analyzed by both methods. Samples of prostates from 10 culture-negative men undergoing prostatectomy for benign hypertrophy, two sets of semen samples from culture-negative sexual partners of 28 women with PID and/or bacterial vaginosis (BV), and ten endometrium-tube sample-pairs from ectopic pregnancies (EPs) were examined by IP only. RESULTS: Seven of the nineteen ovarian specimens tested positive for Chlamydia antigen or deoxyribonucleic acid (DNA) (36%). Of the 10 hypertrophic prostates examined, 4 (40%) were positive. Of the 28 semen samples examined, 10 (35%) tested positive. Tissue samples of 3 cases of EP were positive by IP. CONCLUSIONS: 1. C. trachomatis antigen and nucleic acid can be frequently demonstrated in asymptomatic, culture-negative men and women with chronic infection. 2. Chlamydia antigens may have an etiologic role in benign prostate hypertrophy and EP. 3. Antigenic material may be sexually transmissible. 4. IP and ISH identify temporarily inactive bacteria that may continue to act as immunostimulants and potentially reactivate as Chlamydia infection. PMID- 10836253 TI - Analysis of mRNA levels for the MHC class I-like molecules CD1 and FcRn in preimplantation mouse embryos. AB - PROBLEM: Major histocompatibility complex (MHC) antigens expressed on preimplantation embryos are important for the control of development, reproduction, and allo-recognition of the embryo by the mother. Four types of MHC class I and MHC class I-like antigens have recently been defined: class Ia, class Ib, class Ic, and class Id, based on their similar three-dimensional protein structures. Class Ia and class Ib antigens are encoded in the MHC, whereas class Ic and class Id antigens are encoded by genes on other chromosomes. Both class Ia and class Ib MHC antigens are expressed on preimplantation mouse embryos. The function of the class Ia antigens on embryos is unknown, but the function of one class Ib antigen, Qa-2, the product of the Ped gene, has been found to control the rate of early cleavage division and subsequent embryo survival. The expression of class Ic and class Id antigens on preimplantation embryos has not yet been evaluated. In the present study, we report the analysis of mRNA expression of two class Id genes, CD1 and FcRn, in preimplantation mouse embryos. METHOD OF STUDY: A reverse transcription-polymerase chain reaction (RT-PCR) assay was performed to analyze mRNA levels for CD1 and FcRn in 1-cell, 2-cell, 8-cell, and blastocyst stage embryos from C57BL/6 mice. RESULTS: No expression of CD1 mRNA was found in any of the preimplantation embryos tested. As a by-product of this study, we found a mistake in the published sequence of the mouse CD1 gene: nucleotide 746 in the cDNA is a G not a C. This base change is in a site recognized by the restriction enzyme PstI, thereby eliminating a PstI cleavage site. Expression of mRNA for FcRn was found in all preimplantation stages tested. Higher levels of mRNA for FcRn were detectable in 2-cell and 8-cell embryos compared to 1-cell and blastocyst stage embryos. CONCLUSION: This study shows that mRNA for FcRn but not for CD1 is found in preimplantation mouse embryos. PMID- 10836252 TI - Early pregnancy factor is required at two important stages of embryonic development in the mouse. AB - PROBLEM: The importance of early pregnancy factor (EPF) at the pre-implantation stage of development (days 1-3 post-coitum [p.c.]) has been previously established in this laboratory. However, the role of EPF at the implantation stage (days 4.5-5 p.c.) has not been determined. This present study therefore investigates the role of EPF at this important developmental stage, both in vivo and in vitro. METHOD OF STUDY: Mated mice were passively immunized with anti-EPF antibodies at the peri-implantation stage (days 3.5-4 p.c.) and embryo implantation recorded. Parallel studies were conducted in vitro, where the effect of anti-EPF antibodies on trophoblast outgrowth of blastocysts was determined. RESULTS: Administration of anti-EPF antibodies in vivo at the peri-implantation stage of development resulted in failure of embryos to implant. Similarly, trophoblastic outgrowth of blastocysts was adversely affected in the presence of anti-EPF antibodies. CONCLUSIONS: These results, together with previous findings that anti-EPF antibodies retard embryonic development when administered at the early pre-implantation stage, clearly demonstrate that EPF is required by the embryo at two important developmental stages- the one-two-cell stage and the peri implantation stage. PMID- 10836254 TI - Fas-mediated apoptosis of mouse embryo stem cells: its role during embryonic development. AB - PROBLEM: Fas antigen (APO-1/CD95) can regulate the activity of various cells during adulthood. This study aimed at determining whether Fas may also be involved in the regulation of very early events such as the embryo preimplantation stage. METHOD OF STUDY: We used mouse embryo stem (ES) cell line as a model for testing the effect of Fas crosslinking upon anti-Fas monoclonal antibody (MoAb) treatment. In addition, this treatment was also applied to in vitro mouse-embryo culture. RESULTS: Flow-cytometry analysis of cultured ES cells demonstrated an increase in Fas expression. unchanged in the presence of mouse interleukin-2, while greatly upregulated in the presence of lipopolysaccharide (LPS). As determined by various means, ES cells may undergo a Fas-mediated apoptosis, slightly but significantly intensified by the addition of LPS to cell cultures. We also report that anti-Fas MoAb directly inhibited two-cell stage mouse-embryo (preimplantation) development in in-vitro culture conditions. CONCLUSION: These data suggest a novel mechanism controlling the regulation of physiological cell turnover as well as blastocyst implantation in early embryo development. PMID- 10836255 TI - Comparative evaluation of rumen-protected fat, coconut oil and various oilseeds supplemented to fattening bulls. 1. Effects on growth, carcass and meat quality. AB - Growth performance, carcass and meat quality were determined in 36 fattening Brown Swiss bulls fed with maize silage-hay-concentrate based rations supplemented with fats and various oilseeds. The concentrate diet in the control group contained only barley and soybean meal, while the treated groups included, as partial replacement of the concentrate, rumen-protected crystalline fat, coconut oil, whole crushed rapeseed, sunflower seed or linseed, providing additional 3% fat of total ration dry matter. Animals were housed in groups of six with one representative of each treatment in a monofactorial design and feed was offered using transponder controlled equipment. Daily gains were similar for control, protected fat, rapeseed and linseed treatments (1240 g/d on average), but were lower (P < 0.05) with sunflower seed (1135 g/d) and coconut oil (1038 g/d). Corresponding differences (P < 0.05) in carcass weights were observed. All fat supplemented groups had reduced rumen fluid protozoa counts (P < 0.05). Carcasses tended to be leaner with the fat supplements. Mostly no significant effects on other carcass quality (dressing percentage, conformation score) and meat quality traits (final pH, cooking loss, shear forces) as well as composition (dry matter, fat, collagen) occurred. Consequently, rumen protected fat and some oilseeds can be recommended to be fed to growing cattle as energy sources. PMID- 10836256 TI - Comparative evaluation of rumen-protected fat, coconut oil and various oilseeds supplemented to fattening bulls. 2. Effects on composition and oxidative stability of adipose tissues. AB - The effects of five different dietary fat supplements on fatty acid composition and oxidative stability of subcutaneous and kidney fat were evaluated in 36 Brown Swiss bulls and compared to a low fat diet in a monofactorial design. The following fat supplements were provided as additional fat at 30 g per kg feed dry matter: crystalline rumen-protected fat, coconut oil, and three types of crushed whole oilseeds (rapeseed, sunflower seed and linseed). Adipose tissues reflected differences (P < 0.05) in dietary fatty acid composition although to a lower extent. Using protected fat, which contained elevated levels of trans fatty acids, and sunflower seed, containing a high proportion of linoleic acid, significantly increased C18:1 trans fatty acid proportion in the adipose tissues. The use of sunflower seed increased conjugated linoleic acid. The oilseeds resulted in lower amounts of C16:0 in favour of C18:0. Except for linseed, all fat supplemented groups improved oxidative stability of adipose tissues as compared with control. This was explained by lower proportions of unsaturated fatty acids in adipose tissue (protected fat), by elevated alpha-tocopherol contents (rapeseed, sunflower seed) or by a combination of both (coconut oil). Fat colour remained unaffected by treatments. Compared to other fat supplements oilseeds, especially sunflower seed and rapeseed, can therefore be recommended to be fed to bulls in order to increase the proportions of C18 unsaturated fatty acids in adipose tissues and to maintain or improve oxidative stability. PMID- 10836257 TI - Activity of supplemental enzymes and their effect on nutrient utilization and growth performance of growing chickens as affected by pelleting temperature. AB - Activity of supplemental enzymes in a barley-soybean-maize based diet at 60, 75 and 90 degrees C pelleting temperatures was studied using feed viscosity, in vitro enzyme activity and broiler performance data. High pelleting temperatures increased feed viscosity but supplemented enzymes reduced the viscosity at all three temperatures levels by 11, 14 and 17%, respectively. Water intake and losses in excreta of birds were found to be affected by feed viscosity. Activity of cellulase enzyme, measured using the radial diffusion method, was unaffected at 60 and 75 degrees C, but reduced by 73% in feed processed at 90 degrees C. Enzymes increased the weight gain of broilers by 11.1% at 90 degrees C, but no effect could be seen at low pelleting temperatures possibly due to high dietary protein and energy contents. Feed intake was unaffected by enzymes. Birds consumed 6% more feed and grew 9% faster when the pelleting temperature was increased from 60 to 75 degrees C. Reduced feed intake and daily weight gain observed at 90 degrees C could be fully compensated by the enzyme supplementation. High pelleting temperature reduced energy metabolizability (3.2%) and nitrogen utilization (4%) but enzyme almost compensated them (by 3.3% and 2.6%, respectively). No interaction could be detected between the pelleting temperatures and enzymes. It is concluded that pelleting temperatures as high as 90 degrees C drastically reduce cellulase activity, energy and nitrogen utilization thus lowering broiler performance. Either the remaining activity of cellulase or other thermostable enzymes can prevent the losses. PMID- 10836258 TI - Blood cells and plasma proteins of chickens fed a diet supplemented with (1- >3),(1-->6)-beta-D-glucan and enrofloxacin. AB - The effects of (1-->3),(1-->6)-beta-D-glucan from Saccharomyces cerevisiae and of the fluochinolone enrofloxacin were studied on red and white blood cells and plasma proteins of growing chickens up to the 35th day of life. The prominent findings within the leukocyte population on a per cent scale are: (i) increase of leukocyte count; increase of neutrophils and decrease of lymphocytes in the control and in the antibiotic group from day 17 to day 35; (ii) a minor decrease of neutrophils and no change of lymphocytes in the glucan group; (iii) the monocytes increase from 2.5 +/- 1.8% to 6.5 +/- 7.6% in the glucan group; (iv) the basophils increase in the control group and scale down in the other groups from day 17 to day 35. The total count of leukocytes increases in the controls and in the glucan group. The total protein content of blood plasma, beta-globulin and gamma-globulin increase and the albumin-globulin-ratio and alpha-globulin decline during chickens growth. These changes are most prominent in the glucan group. The haemoglobin concentration shows in all three dietary groups a highly significant increase from day 17 to day 35 by about 17 to 27 per cent; no changes are seen in packed cell volume and number of erythrocytes per litre blood. PMID- 10836259 TI - Endogenous N-losses in broilers estimated by a [15N]-isotope dilution technique: effect of dietary fat type and xylanase addition. AB - Male broilers were given a low protein diet (15.5% CP) spiked with [15N]H4HCO3 from day 12 to day 18 of age to label the endogenous N-constituents. Experimental diets were subsequently fed from day 19 to day 24 of age and consisted of a rye based diet (56% dietary inclusion) which contained either 10% soya oil (S) or 10% beef tallow (T), each of which was either unsupplemented (-) or supplemented (+) with a xylanase containing enzyme preparation (2700 IU/kg at pH 5.3). [15N]-atom percent excess (APE) of excreta, faeces and urine were monitored on a daily basis during both experimental periods. Furthermore, APE was measured in various tissues at the end of the experiment. The APE of urine on the last day of the experiment was between the APE of the pancreas and that of the jejunal tissue, an observation which supported the usefulness of using urinary APE as an indicator for the endogenous N-pool. Endogenous N-proportions were estimated by an isotope dilution technique at the end of the experiment by examination of the ratio of APE in faeces and urine. The endogenous N-proportion in the faeces was greatest in birds receiving the T(-) diet. The proportions were 0.321, 0.319, 0.451 and 0.289 in S(-), S(+), T(-) and T(+) fed groups, respectively. Xylanase addition reduced endogenous N-proportion, a factor which was used to correct apparent crude protein digestibility (85.6, 86.2, 84.3 and 88.5% in S(-), S(+), T(-) and T(+) fed birds, respectively) for endogenous losses resulting in almost equal true digestibilities of crude protein for all treatments (90.3, 90.6, 90.4 and 91.5%). The amounts of endogenous N in faces were estimated to be 87, 69, 244 and 81 mg per day per kg0.67 body weight in S(-), S(+), T(-) and T(+) fed birds, respectively. It was concluded that xylanase supplementation of a rye based broiler diet does not change endogenous N-secretions when the supplemental fat is soya oil. However, addition of tallow rather than soya oil increased these N losses significantly, an effect which was reversed by xylanase addition. PMID- 10836260 TI - RNA bound to a solid phase can select an amino acid and facilitate subsequent amide bond formation. AB - Polyadenylic acid (Poly A) immobilised on silica gel stereoselectively binds L lysine from dilute aqueous solution. When the resulting complex is suspended in an organic solvent and liquid ammonia added, lysinamide is formed. PMID- 10836261 TI - The stability of some selected amino acids under attempted redox constrained hydrothermal conditions. AB - In order to evaluate the stability of aspartic acid, serine, leucine, and alanine under redox buffered hydrothermal conditions, a series of experiments have been performed. The pyrite-pyrrhotite-magnetite (PPM) mineral assemblage was used in the experimental systems in order to constrain the oxygen fugacity. Likewise, the K-feldspar-muscovite-quartz (KMQ) assemblage was added to control the hydrogen ion activity during the experiments. The purpose was to compare the relative stabilities in buffered and unbuffered experiments. The experiments were conducted at 200 degrees C and 50 bar in Teflon coated autoclaves. Glycine, which was not present initially, started to appear at an early stage in the experimental systems and is believed to be the result of decomposition of serine. Similarly, the increase in relative abundance of alanine is likely to be the result of decomposition of serine. Decomposition rates of leucine, alanine and aspartic acid were found to be lower in experiments containing the redox buffer assemblage pyrite-pyrrhotite-magnetite than in non-redox buffered experiments. The decomposition rate of serine was higher in buffered experiments, which indicates that a transformation pathway via dehydration of serine to dehydroalanine followed by reduction to alanine is promoted by reducing conditions. PMID- 10836262 TI - Inhibition of rare earth catalytic activity by proteins. AB - Catalytic action of rare earth element, Ce(IV) to hydrolyze phosphomonoester bonds was confirmed. This effect was considered to suppress abiotic synthesis of nucleotides and nucleic acids in the primitive sea, and hence the origin of life. However, we found that the presence of proteins, especially albumin, strongly inhibited the catalytic action of Ce(IV). This finding was supported by preferential binding of rare earth elements (REEs) to proteins which was revealed using the radioisotopes of these REEs. Consequently, if a large amount of proteins was synthesized in the primitive sea, abiotic synthesis of phosphomonoester compounds, and hence nucleic acids, might have been possible. PMID- 10836263 TI - Sugars as the optimal biosynthetic carbon substrate of aqueous life throughout the universe. AB - Our previous analysis of the energetics of metabolism showed that both the biosynthesis of amino acids and lipids from sugars, and the fermentation of organic substrates, were energetically driven by electron transfer reactions resulting in carbon redox disproportionation (Weber, 1997). Redox disproportionation--the spontaneous (energetically favorable) direction of carbon group transformation in biosynthesis--is brought about and driven by the energetically downhill transfer of electron pairs from more oxidized carbon groups (with lower half-cell reduction potentials) to more reduced carbon groups (with higher half-cell reduction potentials). In this report, we compare the redox and kinetic properties of carbon groups in order to evaluate the relative biosynthetic capability of organic substrates, and to identify the optimal biosubstrate. This analysis revealed that sugars (monocarbonyl alditols) are the optimal biosynthetic substrate because they contain the maximum number of biosynthetically useful high energy electrons/carbon atom while still containing a single carbonyl group needed to kinetically facilitate their conversion to useful biosynthetic intermediates. This conclusion applies to aqueous life throughout the Universe because it is based on invariant aqueous carbon chemistry -primarily, the universal reduction potentials of carbon groups. PMID- 10836264 TI - Polyphosphorylation and non-enzymatic template-directed ligation of oligonucleotides. AB - Oligonucleotide 5'-polyphosphates are formed under potentially prebiotic conditions from oligonucleotide 5'-phosphates and sodium trimetaphosphate. Oligonucleotides activated as polyphosphates undergo template-directed ligation. We believe that these reactions could have produced longer oligonucleotide products from shorter substrates under prebiotic conditions. PMID- 10836265 TI - The possible role of volcanic aquifers in prebiologic genesis of organic compounds and RNA. AB - In a volcanic aquifer, a wide range of physical and chemical conditions are not merely possible, but to be expected: relatively oxidizing and reducing environments both are present; hot and moderate temperatures can be expected; distillation and reflux conditions are probable to allow concentration of reactants, stimulation of reaction and fractionation of isotopes; apatite, hydroxides, clays and sulfide minerals are present to act as chromatographic media for separating compounds, to serve as catalytic surfaces and to provide potential energy sources; supersaturated precipitation of optically active crystals is reasonable, allowing for chromatographic separation of racemic mixtures by the resulting fixed chiral phase; and saturated and unsaturated conditions both are present for promotion of constructive reactions and inhibiting destructive hydrolysis reactions. Because the multitude of physical chemical environments makes the setting robust with respect to circumventing commonly identified problems in origin-of-life theories, even if objections to details proposed herein are identified, the setting is favorable for devising alternatives. This paper describes a theory for the genesis of organic compounds, including RNA, in the mixing zone of juvenile and meteoric waters above a leaky semi-confined aquifer. Starting with basic reactants for best-guess conditions on Archean Earth, parallel sequences of specific reactions are proposed that culminate with RNA oligonucleotides, key molecules in contemporary life. All proposed reactions, or close analogues, are experimentally confirmed and all are set in plausible Archean conditions. Calculations indicate that the proposed reactions would yield C isotopic compositions that are consistent with observed biologic C. PMID- 10836266 TI - Self-programmable, self-assembling two-dimensional genetic matter. AB - Putative two-dimensional coding systems can be constructed from aqueous solutions of purine and pyrimidine nucleic acid bases evaporated at moderate temperatures on the surfaces of inorganic solids. The resultant structures are monolayers which are formed spontaneously by molecular self-assembly and they have been observed with molecular resolution by scanning tunnelling microscopy (STM). When formed from solutions of a single base, the monolayers of adenine and uracil have crystalline characteristics and the STM images can be interpreted in terms of the geometrical placement of planar arranged molecules that interact laterally by intermolecular hydrogen bonding. When formed from solutions containing a mixture of adenine and uracil, the monolayers have aperiodic structures. Small crystalline domains within these monolayers can be interpreted in terms of the single phase configurations of the molecules and the remaining aperiodic structures can presumably be interpreted, geometrically, in terms of the 21 theoretically possible adenine-adenine, uracil-uracil and adenine-uracil hydrogen bonding interactions. We propose that combinatorial arrangements of planar arranged purine and pyrimidine bases could provide the necessary complexity to act as a primitive genetic mechanism and may have relevance to the origin of life. PMID- 10836268 TI - Efficacy of chlorhexidine against some strains of cultured and clinically isolated microorganisms. AB - The efficacy of chlorhexidine digluconate was determined against some strains of collected and clinically isolated bacteria and fungi. The efficacy was evaluated either by calculating a minimum inhibitory concentration (MIC) or by efficacy trials according to the guidelines of the European Committee for Standardization. The MIC values of chlorhexidine for Staphylococcus aureus, Microsporum gypseum, Microsporum canis and Trichophyton mentagrophytes were 0.625 microg/ml, 12.5 microg/ml, 50 microg/ml and 6.25 microg/ml, respectively. The in vitro efficacy of chlorhexidine was higher against ATCC strains of S. aureus and P. aeruginosa (0.5 mg/ml for 5 min and 0.5 mg/ml for 10 min, respectively) than against clinical isolates (0.5 mg/ml for 15 min and 1 mg/ml for 10 min, respectively). The antiseptic activity of aqueous solutions of chlorhexidine against spores of Bacillus subtilis, Bacillis sfericus and Clostridium perfringens required longer contact times than against the vegetative forms. Nevertheless, 5 mg/ml of chlorhexidine in water-ethanol 20:80 v/v was totally effective against the vegetative forms or spores of these microorganisms. PMID- 10836267 TI - Effect of immunosuppression on the clinicopathological changes in experimental zygomycosis in rabbits. AB - A study was undertaken to investigate the effect of immunosuppression by cyclophosphamide or methylprednisolone on the clinicopathological alterations in respiratory absidiosis in rabbits. Infected rabbits showed respiratory distress that was more severe in immunosuppressed groups. Leukocytosis due to neutrophilia was observed in the non-immunosuppressed group in the initial stages, whereas leukopenia was observed in both the immunosuppressed groups initially, owing to polymorphopenia in the cyclophosphamide-treated group and to lymphopenia in the methylprednisolone-treated group, followed by leukocytosis in both groups. Total serum proteins increased significantly in the non-immunosuppressed group but were significantly decreased in the immunosuppressed groups. Serum creatinine increased significantly in all the infected groups from 20 days post inoculation (DPI) onwards. Blood urea nitrogen increased significantly in the initial stages only in the methylprednisolone-treated group. AST and ALT also showed significant increases in the infected animals. Total serum immunoglobulins and circulating immune complexes increased gradually in all three infected groups, except for an initial significant drop in the immunosuppressed rabbits. Re-isolation of fungus was only achieved from the lungs of infected rabbits up to 15 DPI in the non immunosuppressed group and 30 DPI in the immunosuppressed groups. Pathological lesions in all the infected groups were found mainly in the lungs and consisted of pyogranulomas. The lesions were most severe in the cyclophosphamide-treated group and least severe in the non-immunosuppressed group. PMID- 10836269 TI - Hydrocortisone concentrations in post-race urine from horses. AB - As hydrocortisone is an endogenous substance, it is first necessary to establish its normal concentrations so as to be able to control its use in racing animals. This study was designed to establish the hydrocortisone concentrations in post race urine samples of horses racing in Brazil and also to evaluate the results in relation to the international threshold set for this drug. Urine samples were analysed by HPLC-UV. The results were evaluated according to the concentration range as well as sex and time of sample collection (afternoon or evening races). The results showed a high degree of variation in the concentrations of hydrocortisone in the urine (93 +/- 69 ng/ml). The maximum concentration observed was 646 ng/ml, although only a few horses (around 1%) showed levels within the range 500-650 ng/ml, 91% being in the range 0-150 ng/ml. The data suggested a normal distribution curve. Statistical analysis showed no significant influence of sex or time of sample collection. PMID- 10836270 TI - A survey of the literature (1995-1999) on the kinetics of drugs in camels (Camelus dromedarius). AB - Recent publications dealing mainly with the kinetics of antiparasitic and antibacterial agents, NSAIDs, and other drugs in camels are briefly reviewed. The kinetic data for most of these drugs indicated that they have longer absorption and elimination half-lives and slower systemic clearance in the camel compared to other animals. This corroborates earlier reports that suggested that the activities of drug-metabolizing enzymes and the capacity to biotransform and eliminate xenobiotics is lower in camels than in other ruminants. There is a clear need to establish basic kinetic data for the camel in order to avoid extrapolation of drug dosage regimens and withdrawal times from data for other animals, as this may result in irrational use of drugs in camels. PMID- 10836271 TI - A comparison of methods for measuring the antibody response in mice and cattle following vaccination against foot and mouth disease. AB - We present a comparison of methods for evaluating the potency of foot and mouth disease vaccine in the laboratory. The anti-FMDV antibodies (Ab) in vaccinated mice were tested by liquid phase (lp) ELISA, solid phase (sp) ELISA and virus neutralization (VN), and were compared with the Ab titres detected by lpELISA, which is the official test in Argentina for testing the potency of FMD vaccines and protection against a virulent challenge in cattle. The results demonstrated that it is possible to relate the Ab levels induced in vaccinated mice with both the Ab and protective responses elicited in cattle. Furthermore, it was found that the anti-FMDV Ab titres in mice detected by lpELISA 14 days after vaccination should be an accurate parameter for predicting the results of the challenge test in cattle. Thus, this test in mice appears to be an inexpensive and rapid alternative for testing FMD vaccines in cattle. PMID- 10836272 TI - Pathotyping of Newcastle disease viruses by RT-PCR and restriction enzyme analysis. AB - The technique of RT-PCR and restriction enzyme analysis was standardized to detect and differentiate Newcastle disease viruses. Digestion of RT-PCR amplified, F gene sequences encoding for the cleavage activation sites of fusion protein with restriction enzymes AluI, BglI, HaeIII, HinfI, HhaI, RsaI, StyI and TaqI was carried out in order to characterize Newcastle disease viruses of varying pathogenicity. Restriction enzyme digestion of the amplicons by BglI and HhaI could group eight viruses, both field isolates and known vaccine strains, into lentogenic, mesogenic and velogenic pathotypes. By employing this technique directly on a clinical sample, Newcastle disease virus of the lentogenic pathotype could be detected. PMID- 10836273 TI - Effect of vaccination with Brucella abortus strain RB51 on heifers and pregnant cattle. AB - Thirteen cows, which had been vaccinated as calves with strain 19, were revaccinated twice or three times as adults with I x 109 cfu of B. abortus strain RB51. Their serological responses following adult vaccination remained negative to conventional brucellosis surveillance tests. Vaccination with strain RB51 during the eighth month of pregnancy did not induce abortion, although strain RB51 was recovered from milk for up to 69 days after vaccination. In a parallel study, thirteen 8- to 10-month-old heifers were vaccinated as calves with 109 cfu of strain RB51. The heifers remained seronegative to conventional brucellosis surveillance tests but antibody responses to RB51 could be demonstrated using an indirect ELISA. This study showed that multiple vaccination with strain RB51 did not induce seroconversion to brucellosis surveillance tests. In addition, this study suggests that 109 cfu of strain RB51 is safe for use in pregnant cattle. PMID- 10836274 TI - Ultrastructural observation of nasal and pulmonary intracellular Pasteurella multocida A:3 in rabbits. AB - Sixteen 8- to 9-week-old Pasteurella multocida-free rabbits were divided into two equal groups. Eight rabbits in one group were inoculated intranasally with P. multoida type A:3. The other eight were inoculated intranasally with phosphate buffered saline and used as controls. Nasal swabs taken before and after inoculation were cultured for bacterial isolation. Post-mortem nasal swabs and lung samples were cultured for bacteriological isolation. Nasal mucosa and lung samples were collected and processed for transmission electron microscopy. Pasteurella multocida was isolated from the nasal cavity of all infected rabbits and from the lungs of four infected rabbits. Degenerative ultrastructural changes in epithelial cells and endothelial cells were seen in the infected rabbits. Deciliation of the ciliated epithelium and hyperplasia of the goblet cells in the nasal mucosa were noted. Thickening of the alveolar septa due to hyperplasia of type II pneumocytes, swelling of the endothelial lining of capillaries and infiltration of inflammatory cells were also observed. Intracellular invasion of the nasal epithelial cells and of type II pneumocytes by the organism was observed. Coccobacilli were observed in membrane-bound vacuoles in the cytoplasm of these cells. The vacuoles were adjacent to the host-cell mitochondria and some of these vacuoles appeared to be fused to the mitochondrial membrane. Some type I pneumocytes with intracellular membrane-bound vacuoles containing bacterial cells showed protrusions, which appeared to detach into the alveolar lumina. These results indicated that P. multocida serotype A:3 in rabbits can invade the epithelial cell and cause structural changes in the interstitium, epithelium and endothelium. Heterophils and macrophages appear to play important roles in tissue injury. PMID- 10836275 TI - Evaluation of the efficacy of atovaquone alone or in combination with azithromycin against acute murine toxoplasmosis. AB - Mice were infected intraperitoneally with 10,000 tachyzoites of Toxoplasma gondii (RH) strain and, 24 h later, were treated orally for 10 days with atovaquone and azithromycin, either alone or in combination. Evaluation of the efficacy of the drugs was performed by microscopic examination of smears prepared from the organs of the mice, and by subinoculation of visceral and brain suspensions from surviving mice into healthy mice at the end of the experiments. It was found that 58%, 83% and 100% of the mice survived after administration of 75, 150 or 200 mg/kg per day of azithromycin, respectively. Moreover, 8%, 17% and 25% of the mice survived after treatment with atovaquone at 20, 50 or 100 mg/kg per day, respectively. No synergistic or additive effects of combinations of atovaquone and azithromycin were observed. However, azithromycin did not eradicate the parasite from the brain and viscera of the infected mice, whereas atovaquone at 20, 50 and 100 mg/kg per day removed the parasite from viscera and at 100 mg/kg per day eradicated the parasite from the brain of infected mice. The combinations of atovaquone and azithromycin failed to completely eradicate the parasite from the brain and viscera of infected mice. PMID- 10836277 TI - Disposition kinetics and urinary excretion of pefloxacin after intravenous injection in crossbred calves. AB - The disposition kinetics and urinary excretion of pefloxacin after a single intravenous administration of 5 mg/kg were investigated in crossbred calves and an appropriate dosage regimen was calculated. At 1 min after injection, the concentration of pefloxacin in the plasma was 18.95 +/- 0.892 microg/ml, which declined to 0.13 +/- 0.02 microg/ml at 10 h. The pefloxacin was rapidly distributed from the blood to the tissue compartment as shown by the high values for the initial distribution coefficient, alpha (12.1 +/- 1.21 h-1) and the constant for the rate of transfer of drug from the central to the peripheral compartment, K12 (8.49 +/- 0.99 h ). The elimination half-life and volume of distribution were 2.21+/- 0.111 h and 1.44 +/- 0.084 L/kg, respectively. The total body clearance (ClB) and the ratio of the drug present in the peripheral to that in the central compartment (P/C ratio) were 0.454 +/- 0.026 L/kg h) and 5.52 +/- 0.519, respectively. On the basis of the pharmacokinetic parameters obtained in the present study, an appropriate intravenous dosage regimen for pefloxacin in cattle for most of the bacteria sensitive to it would be 6.4 mg/kg repeated at 12 h intervals. PMID- 10836276 TI - Acute and subacute metabolic and endocrine effects of clenbuterol in female pigs. AB - The aim of the study was to assess the relationship between acute and subacute metabolic and endocrine effects after intravenous administration of the beta2 adrenergic agonist clenbuterol in a growth-promoting dose to female pigs. Acute metabolic and endocrine effects were assessed by measuring the blood glucose, serum insulin and nonesterified fatty acid (NEFA) concentrations during 300 min after a single administration of clenbuterol. Significantly higher serum insulin and NEFA concentrations (19.90 +/- 2.50 microU/ml, p<0.01, and 0.69 +/- 0.04 mmol/L, p<0.001, respectively) were measured 30 min after the preprandial administration of clenbuterol in female pigs. Over the same period, the levels of blood glucose (4.42 +/- 0.30 mmol/L) showed no difference from those of control pigs. The postprandial serum NEFA concentration decreased moderately during 210 min after feeding. Postprandial blood glucose and insulin concentrations increased and reached maximal levels 120 min after clenbuterol administration (10.91 +/- 0.60 mmol/L and 85.22 +/- 7.24 microU/ml, respectively), and returned to basal levels at 300 min (4.20 +/- 0.21 mmol/L and 7.75 +/- 1.60 microU/ml, respectively) after the administration of clenbuterol. Subacute metabolic and endocrine effects were assessed by measuring the blood glucose, serum insulin and NEFA concentrations for 21 days after the repeated doses of clenbuterol. In addition, the influence of clenbuterol administration on the endocrine regulation of the onset of the next expected oestrus in female pigs was assessed by measuring their serum 17beta-oestradiol and progesterone concentrations. Blood glucose, serum insulin and NEFA concentrations after the last administration of clenbuterol did not differ significantly from those in control animals. The onset of the next expected oestrus occurred regularly without any significant difference in serum 17beta-oestradiol or progesterone concentrations between the treated (9.83 +/- 2.60 pg/ml and 0.15 +/- 0.03 ng/ml) and control pigs (8.52 +/- 2.70 pg/ml and 0.25 +/- 0.06 ng/ml). The study results suggest the duration of intravenous administration of clenbuterol in a growth-promoting dose necessary to influence the metabolic and endocrine activities in female pigs. PMID- 10836278 TI - An apparent outbreak of cutaneous papillomatosis in merino sheep in patagonia, Argentina. AB - A retrospective study was performed on skin samples from an outbreak of cutaneous papillomatosis in Merino sheep that occurred in 1995. The samples were processed for routine histology, electron microscopy and immunocytochemistry for papilloma viruses. Particles of approximately 55 nm diameter were found in some nuclei of the stratum granulosum cells, while immunocytochemistry gave positive staining of cell nuclei in this layer. This study confirms that papillomas associated with papillomaviruses occur in sheep in Patagonia. PMID- 10836279 TI - Antigenic characterization of rabies virus isolates from vaccinated dogs in plateau state, Nigeria. AB - Rabies isolates (genotype 1 lyssaviruses) from vaccinated dogs that died of rabies infection in the Plateau area of Nigeria were characterized using monoclonal antibodies (MAbs). The isolates were examined for rabies (genotype 1) and rabies-related (genotypes 2, 3 and 4) viruses by the indirect fluorescent antibody test carried out with MAb 502-2, which recognizes the nucleocapsid protein of all known lyssaviruses, and with MAb 422-5, which identifies only rabies-related viruses. All three isolates showed positive immunofluorescence with MAb 502-2 and were negative with MAb 422-5, indicating that they were all rabies (genotype 1) viruses. Characterization with a panel of 36 anti nucleocapsid monoclonal antibodies showed that all three isolates reacted positively with 35 of the anti-nucleocapsid MAbs, including MAb 102-27 and MAb 377-7. Characterization using a panel of 44 anti-glycoprotein MAbs differentiated the isolates sharply from LEP Flury and PM vaccine viruses. The pattern of anti glycoprotein reactivity of the isolates showed them to belong to one distinct viral subtype, except for a minor variation in one isolate that was not neutralized by MAb 1101-3. None of the three isolates was identified as the Flury low egg passage (LEP) vaccine strain used for vaccinating dogs in Nigeria. In fact, all the three isolates had the typical pattern of reactivity of isolates from unvaccinated dogs, including MASS 83, a rabies virus isolated in Nigeria and characterized at the Wister Institute before this study. PMID- 10836280 TI - Current aspects of valproate in bipolar disorder. AB - For more than 30 years, lithium has been the drug of choice for the treatment of bipolar disorder. However, it has numerous adverse effects, a relatively slow onset of action, many common drug-drug interactions, and a narrow therapeutic index. Because of these problems, researchers looked for alternative and/or adjunctive treatments in bipolar disorder, focusing on the anticonvulsants carbamazepine and valproate. The existing data of valproate are reviewed pointing out its promises and limitations in psychiatric diseases. Growing data indicate that valproate is a well tolerated and effective agent in bipolar disorder. Controlled studies prove its use in acute mania, often with a rapid onset of action. Open studies suggest that the drug also reduces the frequency and intensity of recurrent manic and depressive episodes over extended periods. Its acute and prophylactic antidepressant effects are probably minor to its antimanic efficacy. Recent data support the specific therapeutic efficacy of valproate in certain subtypes of bipolar illness: rapid cycling variant, mixed mania, bipolar disorder associated with panic attacks, comorbid alcohol or substance abuse, with neurological features or secondary to organic illnesses. These features make valproate interesting as an alternative treatment for patients who generally respond less well to lithium or as a useful adjunct in the treatment of complicated patients who do not respond to single agents. However, further controlled studies are warranted to provide clear guidelines for the treatment with valproate. PMID- 10836281 TI - Long-term safety and efficacy of amisulpride in subchronic or chronic schizophrenia. Amisulpride Study Group. AB - Amisulpride is an atypical antipsychotic with selective affinity for dopamine D2/3 receptors. In this long-term, open, randomised, multicentre trial, patients with chronic or subchronic schizophrenia received amisulpride (n =370) or haloperidol (n = 118) for 12 months. Dosage regimens were flexible (amisulpride 200-800 mg/day, haloperidol 5-20 mg/day). Improvement in mean Brief Psychiatric Rating Scale total score was significantly greater for amisulpride than haloperidol (17.0 versus 12.8, P = 0.01). Positive symptoms (Positive and Negative Syndrome Scale [PANSS] positive) improved in a similar way in each group but amisulpride caused a significantly better improvement in negative symptoms (PANSS negative) (7.1 versus 3.7, P < 0.0001). Improvements in Global Assessment of Functioning (GAF) and Quality of Life Scale (QLS) scores were also significantly greater in the amisulpride group (GAF -20.1 versus -13.6, P = 0.001; QLS -0.64 versus -0.30, P = 0.02). Adverse events were mainly psychiatric in nature, and occurred with similar frequency in each group (amisulpride 254/370, 69%; haloperidol 82/118, 70%). Extrapyramidal symptoms were more frequent for haloperidol (48/118, 41% versus 96/370, 26% for amisulpride), leading to a greater requirement for antiparkinsonian medication (haloperidol 66/118, 56% versus amisulpride 118/370, 32%). Haloperidol significantly aggravated parkinsonism, akathisia and involuntary movement compared to amisulpride. The overall incidence of endocrine events was comparable between groups (4% for amisulpride, 3% for haloperidol). Maintenance of efficacy was comparable in both treatment groups; 59% of amisulpride patients and 55% of haloperidol patients improved after 1 month of therapy remained improved throughout the study period. Amisulpride is effective following flexible long term administration and significantly improves social functioning and quality of life. PMID- 10836282 TI - Effective open-label treatment of tourette's disorder with olanzapine. AB - Olanzapine is an atypical antipsychotic drug which shows a high antagonistic affinity to the D1, D2 and D4 and the 5-HT2A and 5-HT2c receptors. The goal of our investigation was to assess the efficacy of olanzapine in patients with Tourette's disorder who were either antipsychotic naive or who did not tolerate and/or did not respond to previous antipsychotic treatments in an open-label pilot study. Fourteen patients with a mean (SD 12.4) age of 32.6 years were treated for a period of 6 weeks. Seven patients did not respond to, or did not tolerate, previous neuroleptic treatments and seven patients were antipsychotic naive. All patients received olanzapine in ascending dosage, following a washout period of 1 week. Initial dosage was 10 mg/day with a maximum dosage of 20 mg/day. The Yale Global Tic Severity Scale (YGTSS), Fischer Symptom Check List Neuroleptika and the Clinical Global Impression Severity Scale (CGI) were used. Two of the 14 patients did not complete the investigation. The mean dosage of olanzapine was 15 mg/day (SD 3.3) at day 42 (end of the study). The YGTSS scores and the CGI significantly decreased over the treatment period. The only side effect observed was mild sedation which decreased during the course of the investigation and two patients had weight gain of 3-5 kg with increased appetite. In our study, we found that olanzapine was a safe and effective treatment alternative to other antipsychotics. In order to confirm these preliminary results, double-blind placebo controlled trials are warranted. PMID- 10836283 TI - Increased remission rates with venlafaxine compared with fluoxetine in hospitalized patients with major depression and melancholia. AB - This was a 6-week, double-blind, randomized trial of the efficacy and tolerability of venlafaxine and fluoxetine in 109 patients with major depression and melancholia. Hospitalized and day care patients with DSM-IV major depression and melancholia and a baseline Montgomery-Asberg Depression Rating Scale (MADRS) score of > or = 25 were eligible. The doses were venlafaxine 75 mg/day or fluoxetine 20 mg/day from days 1-4, venlafaxine 150 mg/day or fluoxetine 40 mg/day from days 5-10, and venlafaxine 225 mg/day or fluoxetine 60 mg/day from days 11-42. The intention-to-treat analyses included 55 patients on venlafaxine and 54 on fluoxetine. At the final evaluation, 70% of patients with venlafaxine and 66% with fluoxetine had > or = 50% reduction in the MADRS score, and 70% with venlafaxine and 62% with fluoxetine had a Clinical Global Impression (CGI) score of 1 or 2. A CGI improvement score of 1 was observed in 51% of patients with venlafaxine and 32% with fluoxetine (P = 0.018). A final Hamilton Depression Rating Scale (HAM-D) score < 7 was attained in 41% of venlafaxine-treated and 36% of fluoxetine-treated patients. Overall, 22% of patients in each group discontinued therapy, but only 5% on venlafaxine and 9% on fluoxetine discontinued for adverse events. Nausea was reported in 5.5% of venlafaxine treated patients and 14.8% of fluoxetine-treated patients. Venlafaxine was effective and well tolerated for treating inpatients with major depression and melancholia. Based on remission criteria (HAM-D < 7 or CGI of 1), venlafaxine was superior to fluoxetine. PMID- 10836284 TI - LSD-induced hallucinogen persisting perception disorder treatment with clonidine: an open pilot study. AB - A pilot open study was conducted in order to evaluate the efficacy of clonidine in the treatment of LSD-induced hallucinogen persisting perception disorder (HPPD). Eight patients fulfilled entrance criteria. All complained of HPPD for at least 3 months and were drug free at least 3 months. They received fixed low doses of clonidine, 0.025 mg, three times a day for 2 months. They were evaluated by the Clinical Global Impression Scale (CGI) and a self-report scale on the severity of symptoms (graded 0-5). Patients scored an average of 5.25 (SD = 0.46) on the CGI and 4 on the self-report scale at baseline, indicating marked psychopathology. One patient dropped out at week 3 and a second patient dropped out at week 5. Of the six patients remaining at the end of 2 months, the average CGI score was 2.5 (SD = 0.55) and the self-report scale score was 2, indicating mild symptomatology. LSD-related flashbacks associated with excessive sympathetic nervous activity may be alleviated with clonidine in some patients. PMID- 10836285 TI - Prediction of lithium response using. m-chlorophenylpiperazine challenge test: A preliminary finding. AB - It is important to be able to predict a response to lithium before lithium administration because it usually takes 2 or 3 weeks for lithium to manifest its effect sufficiently. We hypothesized that lithium responders have a high post synaptic serotonergic receptor function, whereas non-responders have low one. This is because it has been suggested that lithium's primary actions on the serotonergic neuron may be presynaptic and several reports have shown that lithium enhances serotonin synthesis and secretion at the presynaptic serotonergic neuron. As a preliminary study, we investigated the relationship between several responses to meta-chlorophenylpiperazine (a serotonergic agonist) reflecting the serotonergic receptor function and the clinical lithium response of 10 patients. No significant relationship between hormone responses and improvement ratios was found. However, there were significant relationships found between 'anger' during the challenge test and the improvement ratio and between 'tension' during the test and the improvement ratio. Thus, it seems that the hormone response to a serotonergic agonist is not useful for predicting lithium response. However, subjective responses such as anger and tension to a serotonergic agonist are possible candidates for predicting lithium response. In any case, further studies with larger numbers of participants are required to investigate whether subjective responses to a serotonergic agonist are useful for predicting lithium response. PMID- 10836286 TI - The efficacy and tolerability of venlafaxine and paroxetine in outpatients with depressive disorder or dysthymia. AB - A 24-week, double-blind, randomized trial was performed to compare the efficacy and tolerability of venlafaxine and paroxetine in patients with major depression or dysthymia. Outpatients aged 18-70 years with a baseline score of 17 on the 21 item Hamilton Depression Rating Scale (HAM-D) were eligible. Patients were randomly assigned to venlafaxine, 37.5 mg, in the morning and evening or paroxetine, 20 mg, in the morning and placebo in the evening, which could be increased to venlafaxine, 75 mg twice daily, or paroxetine, 20 mg twice daily, after 4 weeks. Efficacy was assessed with the 21-item HAM-D, the Montgomery Asberg Rating Scale, the Hamilton Anxiety Rating Scale, and the Clinical Global Impressions Scale. Forty-one patients were randomized to venlafaxine and 43 to paroxetine. At week 6, a response was observed in 55% of patients on venlafaxine and 29% on paroxetine (P = 0.03). At week 12, significantly (P = 0.011) more patients in the venlafaxine group had a HAM-D remission score of 8 or less (59% versus 31%). Discontinuation for any reason occurred in 16 (39%) patients on venlafaxine and 11 (26%) on paroxetine. The most common adverse events were nausea (28%), headache (18%) and dry mouth (15%) with venlafaxine and headache (40%) and constipation (16%) with paroxetine. Venlafaxine was effective and well tolerated for the treatment of patients with mild to moderate depression or dysthymia. A consistently higher proportion of patients had a response or remission on venlafaxine than on paroxetine. PMID- 10836287 TI - Potential use of ipatropium bromide for the treatment of clozapine-induced hypersalivation: a preliminary report. AB - Sialorrhea is reported by 31% of patients taking clozapine. Anticholinergic agents and adrenergic agonists are used for its treatment based on empirical evidence. In the present study, 10 patients who failed to respond to anticholinergic or adrenergic agents received intranasal ipatropium bromide (IPB) to minimize anticholinergic systemic absorption. Intranasal IPB was given to 10 patients for clozapine-induced sialorrhea who failed to respond to benztropine or clonidine. Pre-, post- and 6 month follow-up values were recorded on a single item, 5-point Hypersalivation Rating Scale. The sign test was used for statistical comparison (P < 0.05). Eight patient reported initial improvement in sialorrhea values. Two patients reported no change and two patients discontinued IPB. At 6 months, six patients maintained improvement. Side-effects for IPB were minor. A significant trend was observed in the values pre- and post-treatment with IPB (P < 0.004). Improvement was maintained at 6 month follow-up (P < 0.008). This case series demonstrates the possible utility of intranasal IPB for clozapine-induced sialorrhea. Intranasal IPB lacks significant systemic anticholinergic effects when prescribed along with clozapine. This study shows only qualitative differences in salivation values and large controlled comparative trials are needed. PMID- 10836288 TI - Is selectivity for serotonin uptake associated with a reduced emergence of manic episodes in depressed patients? AB - To determine whether selectivity for serotonin reuptake plays a role in antidepressant-associated mania (AAM), we evaluated the frequency of treatment emergent mania in patients with unipolar depression who received either citalopram, a highly selective serotonin uptake inhibitor, or the adrenergic tetracyclic antidepressants (TTCAs) maproriline and mianserin, or placebo. Data were collected from post-marketing reports of adverse events, three placebo controlled trials and four double-blind comparative trials. Of the total 4,004 depressed patients treated with citalopram (2482 from postmarketing data, 840 from placebo-controlled studies and 682 from TTCAs comparative studies), 25 (0.62%) had manic episodes. The rate of AAM in the comparative trials was significantly lower in the citalopram-treated patients (1/682, 0.15%) than in the TTCA-treated patients (5/389, 1.29%) (P = 0.03). In the placebo-controlled studies, no manic episodes were reported in the patients given placebo, but one manic episode occurred in a citalopram-treated patient (1/840, 0.12%). The citalopram-treated patients in whom AAM developed were significantly older than those in whom it did not (about 10 years, P < 0.001); gender distribution was similar. In conclusion, despite its limitations, our study apparently indicates that citalopram, a highly selective serotonin reuptake inhibitor, is associated with a significantly lower rate of treatment-emergent manic episodes than TTCAs, which have noradrenergic activity, but a similar rate to that reported for less selective SSRIs. PMID- 10836289 TI - Remission of severe tardive dyskinesia in a schizophrenic patient treated with the atypical antipsychotic substance quetiapine. AB - In a single inpatient case study, a schizophrenic patient with tardive dyskinesia after prolonged treatment with typical neuroleptics was treated with the new atypical neuroleptic quetiapine, a dibenzothiazepin-derivative. Within 2 weeks of treatment with quetiapine, symptoms of tardive dyskinesia improved; 10 weeks after starting treatment tardive dyskinesia stopped completely. Over the same period, dopamine D2 receptor occupancy decreased substantially, as measured by IBZM-SPECT after 14 and 77 days of treatment. PMID- 10836290 TI - Ovarian serous borderline tumors: the citadel defended. PMID- 10836291 TI - The pathology of human West Nile Virus infection. AB - West Nile Virus (WNV) was identified by immunohistochemistry (IHC) and polymerase chain reaction (PCR) as the etiologic agent in 4 encephalitis fatalities in New York City in the late summer of 1999. The fatalities occurred in persons with a mean age of 81.5 years, each of whom had underlying medical problems. Cardinal clinical manifestations included fever and profound muscle weakness. Autopsy disclosed encephalitis in 2 instances and meningoencephalitis in the remaining 2. The inflammation was mostly mononuclear and formed microglial nodules and perivascular clusters in the white and gray matter. The brainstem, particularly the medulla, was involved most extensively. In 2 brains, cranial nerve roots had endoneural mononuclear inflammation. In addition, 1 person had acute pancreatitis. Based on our experience, we offer recommendations for the autopsy evaluation of suspected WNV fatalities. PMID- 10836292 TI - Contribution of molecular genetic data to the classification of sarcomas. AB - Many sarcomas are characterized by specific recurrent chromosomal translocations which provide powerful diagnostic tumor markers. Since 1992, the genes involved by almost all of these translocations have been cloned, inaugurating a new era in the study of sarcomas. At the biological level, these chromosomal translocations produce highly specific gene fusions, usually encoding aberrant chimeric transcription factors. Clinically, the correlation of these translocation-derived genetic markers and discrete histopathologic entities has been remarkable. Fusion gene detection has confirmed and refined the nosology of several sarcoma groups. The overall effect has been to strengthen certain pathological concepts rather than to revolutionize. The focus of this brief review is the recent impact that the cytogenetic and molecular detection of these translocations has had on sarcoma diagnosis and classification. PMID- 10836293 TI - Ovarian serous borderline tumors: a critical review of the literature with emphasis on prognostic indicators. AB - BACKGROUND: The behavior of ovarian serous borderline tumors (SBTs) and significance of various prognostic factors are unclear and difficult to evaluate because of inconsistencies and confusion in the literature. Recent studies have suggested that the morphological features of the primary tumor (presence or absence of micropapillary features) and the peritoneal "implants" (presence or absence of invasive features) can reliably subclassify SBTs into benign and malignant types. The aim of the current review was to test two hypotheses. First, that the alleged malignant behavior of SBTs is poorly documented, and second, that the morphological features of the primary ovarian tumors and the associated peritoneal implants are sufficient to separate SBTs into benign and malignant types, thereby obviating the need for the category. METHODS: 245 studies reporting approximately 18,000 patients with borderline ovarian tumors were reviewed. After excluding series that lacked clinical follow-up or were not analyzable for other reasons, there remained 97 reports that included 4,129 patients. In addition to recurrences and survival, we evaluated the type of peritoneal implants, microinvasion, lymph node involvement, late recurrences, and progression to carcinoma, as these features have served as the underpinning of the concept of "borderline malignancy" or "low malignant potential." RESULTS: Among 4,129 patients with SBTs reviewed, the recurrence rate after a mean follow up of 6.7 years was 0.27% per year for stage I tumors, the disease-free survival was 98.2%, and the overall disease-specific survival rate was 99.5%. For patients with advanced-stage tumors, the recurrence rate was 2.4% per year. However, the majority (69%) of reported recurrences were not pathologically documented, and only 26 cases (8.4% of all recurrences) were documented to have recurred from an adequately sampled ovarian tumor. The most reliable prognostic indicator for advanced stage tumors was the type of peritoneal implant. After 7.4 years of follow-up, the survival of patients with noninvasive peritoneal inplants was 95.3%, as compared with 66% for invasive implants (P < .0001). Microinvasion in the primary ovarian tumor was associated with a 100% survival rate at 6.7 years, and lymph node involvement was associated with a 98% survival rate at 6.5 years. The few reported cases of stage IV disease, progression to invasive carcinoma, and very late (>20 years) recurrences were poorly documented. The survival for all stages among approximately 373 patients in 6 prospective randomized trials followed for a mean of 6.7 years was 100%. CONCLUSION: Surgical pathological stage and subclassification of extraovarian disease into invasive and noninvasive implants are the most important prognostic indicators for SBTs. Survival for stage I tumors is virtually 100%. Survival for advanced stage tumors with noninvasive implants is 95.3%, whereas survival for tumors with invasive implants is 66%. Invasive implants behave as carcinomas and are most likely metastatic. The precise nature of so-called noninvasive implants is not clear, but they behave in a benign fashion. The presence of a micropapillary architecture in the primary ovarian tumor is a strong predictor of invasive implants. These data support the recommendation that ovarian tumors with a micropapillary architecture be designated "micropapillary serous carcinomas," and those lacking these features, "atypical proliferative serous tumors." PMID- 10836294 TI - Immunoreactive E-cadherin, alpha-catenin, beta-catenin, and gamma-catenin proteins in hepatocellular carcinoma: relationships with tumor grade, clinicopathologic parameters, and patients' survival. AB - We evaluated the immunohistochemical expression status of E-cadherin, alpha catenin, beta-catenin, and gamma-catenin, and the relationship with tumor grade, clinicopathologic parameters, and patients' survival, in 107 surgically resected hepatocellular carcinoma (HCC), using a semiquantitative scoring system. These molecules were largely located at the cell membrane of HCC cells. Compared with expression in nontumorous liver, E-cadherin showed underexpression, whereas alpha , beta-, and gamma-catenins showed overexpression in most HCC. E-cadherin expression significantly correlated inversely with HCC histological grade, being the highest in well-differentiated HCC. In contrast, alpha-, beta-, and gamma catenins' expression significantly correlated positively with HCC grade, being the highest in poorly differentiated HCC. Significant positive correlations were found between gamma-catenin high expression and capsular invasion or presence of satellite nodules, and between beta-catenin high expression and vascular invasion. Kaplan-Meier examination of patients' survival indicated that HCC patients with underexpression of E-cadherin, alpha-catenin, and gamma-catenin, and patients with overexpression of beta-catenin, had poor survival rates. These results suggest that E-cadherin is downregulated while the 3 catenins are upregulated in HCC, that E-cadherin expression inversely correlates with HCC grade while the 3 catenins' expression positively correlates with HCC grade, and that HCC patients with downregulation of E-cadherin, alpha-catenin, and gamma catenin and HCC patients with upregulation of beta-catenin have poor prognosis. PMID- 10836295 TI - Molecular identification of main cellular lineages as a tool for the classification of gastric cancer. AB - Gastric carcinomas (GC) are heterogeneous tumors comprising variable amounts of cells of different lineage phenotype, including gastric mucous cells (surface- SMC or gland--GMC) and intestinal cells (IC). The evaluation of tumor behavior has classically depended on strictly morphological classifications of tumors. Microsatellite instability (MSI) is frequently detected in GC, but whether MSI affects all gastric cellular lineages or exclusively occurs in unique cellular lineages in GC is not known. The aims of this study were to test a combination of anti-mucin antibodies to classify gastric cancer into predominant cell lineage phenotype and to determine whether MSI in GC is associated with particular cellular tumor phenotypes. Fifty-five GC were immunophenotyped with antibodies specific for SMC, GMC, or IC. DNA was extracted from tumor and non-neoplastic gastric tissues and amplified with 5 microsatellite markers. A mixed cellular pattern was the most frequent phenotype of GC (61%) and was seen in both glandular (63%) and diffuse (58%)-type tumors. No significant difference in the rate of MSI was found in tumors with predominant gastric, intestinal or mixed phenotype. However, tumors with null or low-level expression of cellular lineage differentiation markers displayed MSI more frequently than tumors with high-level expression (40% v 20%). In conclusion, different gastric carcinoma cell lineage patterns can be easily identified with the 3 immunohistochemical markers used in this study. The 3 main cellular lineage components of gastric cancer can be similarly affected by microsatellite instability, consistent with the notion that MSI is an early event in gastric carcinogenesis. PMID- 10836296 TI - Alpha1-antichymotrypsin globules within hepatocytes in patients with chronic hepatitis C and cirrhosis. AB - Alpha1-antichymotrypsin (A1AC) is an acute phase serine protease inhibitor, similar to alpha1-antitrypsin (A1AT) in amino acid sequence. A1AT deficiency is known to be associated with emphysema and cirrhosis; deficiency of serum A1AC has been reported to be associated with emphysema, childhood asthma, and cryptogenic cirrhosis. The hepatocyte globules associated with A1AT deficiency have been well described; A1AC deficiency also has been reported to be associated with hepatocyte globules. The aim of this study was to describe the globules of A1AC and to compare them with A1AT globules. Immunohistochemistry for A1AC and A1AT was performed on liver biopsy specimens from 15 hepatitis C virus (HCV)-positive cirrhotic patients, 14 non-HCV cirrhotic patients, and 12 other patients with chronic hepatitis C but no cirrhosis, all of whom had known serum levels of A1AC; most had known serum levels of A1AT. Five of 15 HCV-positive cirrhotic patients, 1 of 14 non-HCV cirrhotic patients, and 1 of 12 noncirrhotic chronic hepatitis C patients had A1AC globules. Two of 15 HCV-positive cirrhotic patients and 2 of 14 non-HCV cirrhotic patients had A1AT globules. Histologically, the globules of A1AC were similar to those of A1AT but were smaller and fewer; the PAS/D stain was not as helpful for A1AC as it was for A1AT; immunohistochemistry was most useful. There was not a good correlation between serum levels of A1AC and its globules in hepatocytes. A1AC globules should be included in the differential diagnosis of hepatocyte inclusions. PMID- 10836297 TI - Metastatic patterns of prostate cancer: an autopsy study of 1,589 patients. AB - The prognosis of prostate cancer is mainly determined by the presence or absence of metastases. Nevertheless, the metastatic pathways in prostate cancer are not entirely understood. Among 19,316 routine autopsies performed from 1967 to 1995 on men older than 40 years of age, the reports from those 1,589 (8.2%) with prostate cancer were analyzed. Hematogeneous metastases were present in 35% of 1,589 patients with prostate cancer, with most frequent involvement being bone (90%), lung (46%), liver (25%), pleura (21%), and adrenals (13%). Several lines of evidence suggested the existence of a backward metastatic pathway through veins from the prostate to the spine in addition to classical hematogeneous tumor spread via the vena cava. First, there was an inverse relationship between spine and lung metastases, suggesting that metastasis to the spine is independent of lung metastasis. Second, the maximum frequency of spine involvement occurred in smaller tumors (4 to 6 cm) as compared with the maximum spread to lung (6 to 8 cm) and liver (>8 cm), suggesting that spine metastases precede lung and liver metastases in many prostate cancers. Third, there was a gradual decrease in spine involvement from the lumbar to the cervical level (97% v 38%), which is consistent with a subsequent upward metastatic spread along spinal veins after initial lumbar metastasis. The results of this study show that bone, lung, and liver are the most frequent sites of distant prostate cancer metastases. Besides the cava-type of metastasis through lung passage, there are strong arguments for the existence and clinical significance of a backward venous spread to the spine, which is likely to occur early in the metastatic process. PMID- 10836298 TI - A novel flow cytometric steroid hormone receptor assay for paraffin-embedded breast carcinomas: an objective quantification of the steroid hormone receptors and direct correlation to ploidy status and proliferative capacity in a single tube assay. AB - Semiquantitative estimation of steroid hormone receptors by immunohistochemistry applied to paraffin sections is common practice in surgical pathology. Flow cytometric (FCM) analysis of estrogen receptor (ER) and progesterone receptor (PR) levels provides a faster and more objective quantitative assay. However, a major problem in such FCM analyses of solid tumor samples is the admixture of tumor cells with normal epithelial, stromal, and inflammatory cells. The aim of the underlying study was to investigate the applicability of a recently developed multiparameter flow cytometric methodology for the accurate estimation of the fraction of steroid hormone receptor-positive tumor cells and to explore whether this multiparameter approach allows the detection of specific, clinically relevant subsets of tumors, based on a combination of ploidy level, steroid hormone receptor status, and cell cycle characteristics. For this purpose, samples of 42 breast cancer patients, from which routine immunohistochemistry for ER and PR also was available, were analyzed. From each case, a cell suspension was prepared from the paraffin block by applying a heating and short pepsin digestion step to 50-microm-thick sections. These cell suspensions were double immunostained for cytokeratin to identify the epithelial cells, and ER or PR, whereas DNA was quantitatively stained with propidium iodide using an optimized protocol. In the entire group of breast tumors, the percentages of ER- and PR positive cells were registered in the epithelial subfraction, in combination with DNA ploidy and S phase fraction (SPF). A significant correlation was found between the fraction of hormone receptor-positive cells as found by the immunohistochemical and FCM procedures. For ER, a correlation coefficient of r = .87 was found, and for PR r = .62, both P < .0001. It became clear that all the diploid breast tumors had more than 30% tumor cells positive for ER with a SPF lower than 10%, whereas aneuploid tumors contained on average a smaller percentage of steroid hormone receptor-positive cells, and simultaneously an SPF greater than 10%. Our results show that this multiparameter FCM analysis allows an objective and reproducible quantification of the fraction of steroid hormone receptor-positive cells in the relevant epithelial cell compartment in relation to DNA ploidy status and proliferative capacity in a single-tube assay. PMID- 10836299 TI - Dissociated overexpression of cathepsin D and estrogen receptor alpha in preinvasive mammary tumors. AB - The role of estrogen as a promoter agent of sporadic breast cancer has been considered by assaying, in benign breast disease (BBD) and in situ carcinomas (CIS), 2 markers, the estrogen receptor alpha (ERalpha) and cathepsin D (cath-D) involved in estrogen action on mammary tissue. ERalpha and cath-D were assayed by quantitative immunohistochemistry using an image analyzer in 170 lesions of varying histological risk (94 BBD and 76 CIS), and in "normal" glands close to these lesions. The ERalpha level increased significantly in proliferative BBD with atypia (P < .001), in non-high-grade CIS (P < .001), and in adjacent "normal" glands. ERalpha level was decreased in high-grade ductal CIS (DCIS) and also in adjacent "normal" glands. Cath-D level increased in ductal proliferative BBD (P < or = .01) and in high-grade DCIS (P < or = .003), but not in the other lesions. After menopause, ERalpha level was increased (P = .012) but not cath-D level. According to Mac Neman test, the high-grade DCIS were predominantly ERalpha negative and cath-D positive (P = .0017), and the other CIS were predominantly ERalpha positive and cath-D negative (P = .0002). The 2 markers are overexpressed early in premalignant lesions, but independently. This dissociation suggests a branched model of mammary carcinogenesis involving 1 estrogen independent pathway with high cath-D and low ERalpha levels (including high-grade DCIS) and 1 estrogen-dependent pathway, with high ERalpha level (including proliferative BBD with atypia and low-grade DCIS). We propose that ERalpha negative breast cancers may develop directly from high-grade DCIS and that ERalpha assay in preinvasive lesions should be considered in prevention trials with antiestrogens. PMID- 10836300 TI - The delimitation of tubular carcinoma of the breast. AB - Five hundred consecutive breast carcinomas from the first screening round of the Breast Cancer Detection Demonstration Project were studied quantitatively and semiquantitatively for features relevant to the diagnosis of tubular carcinoma. Tubularity was defined as the proportion of tumor cells that were adjacent to an open lumen. Nuclear morphology and mitotic activity were graded 1 to 3, and the presence of apocrine snouts as absent, few, common, or prominent. In plots and statistical cluster analysis, tubular carcinoma appears as part of a continuous spectrum of morphologies and not as a distinct entity. In multivariate analysis, apocrine snouts had no significant association with either nodal status or deaths of breast cancer. Tumors with 70% or greater tubularity by our definition and mitosis and nuclear grades 1 were not associated with either nodal metastases or deaths of breast cancer. The question is raised whether tubular carcinoma at the benign end of a spectrum shades into benign glandular proliferations, with particular reference to microglandular adenosis. A uniform and precise definition of tubularity is needed for the attainment of sufficient collective experience to delimit tubular carcinoma both from more aggressive carcinomas and from benign proliferations. PMID- 10836301 TI - Clinical evidence of distinct subgroups of astrocytic tumors defined by comparative genomic hybridization. AB - In astrocytic tumors, the relationship between genetic pathways and patients' prognoses has not been fully investigated. In our studies of astrocytic tumors using comparative genomic hybridization, the presence of 8q gain was mutually exclusive of 7p gain or amplification. In this study, 45 cases of astrocytic tumor were divided into 3 groups: those with 7p gain, cases with 8q gain, or those with neither; and their clinical course, p53, and epidermal growth factor receptor (EGFR) expressions and proliferative activity were then compared. Of the cases examined, 17 (12 glioblastomas and 5 anaplastic astrocytomas) showed 7p gain. Eleven cases (5 glioblastomas, 2 anaplastic, and 4 low-grade astrocytomas) showed 8q gain. p53 accumulation was observed more frequently in cases with 8q gain than in those with 7p gain. Astrocytic tumors with 8q gain occurred more frequently in younger patients than those with 7p gain. Kaplan-Meier survival rate analysis showed higher survival rates in patients with 8q gain than in those with 7p gain. This tendency also was observed when only patients with malignant glioma were included in the survival analysis. Our results provide evidence for distinct clinical manifestations in astrocytic tumors with 8q and 7p gain. PMID- 10836302 TI - Repeated dedifferentiation of low-grade intraosseous osteosarcoma. AB - Low-grade intraosseous osteosarcoma is an uncommon bone tumor that is characterized by minimum cytological atypism and a much better prognosis than conventional osteosarcoma. This report describes a patient who had a low-grade osteosarcoma that mimicked fibrous dysplasia (FD). The tumor had an area of high grade sarcoma at the initial diagnosis. Ten years after incomplete resection of FD-like tumor, local recurrence with areas of high-grade tumor developed. This case illustrates the potential of dedifferentiation in low-grade intraosseous osteosarcoma. PMID- 10836303 TI - Chronic right-sided myocarditis mimicking arrhythmogenic right ventricular dysplasia. AB - Arrhythmogenic right ventricular dysplasia (ARVD) is a cause of right ventricular heart failure and has been implicated in some cases of sudden death in young adults. It is well known that a large majority of patients with ARVD have histological evidence suggestive of inflammation. Here we report a unique case of chronic myocarditis limited to the right ventricle and right side of the interventricular septum which presented clinically as ARVD. The fact that right sided myocarditis can clinically mimic the genetic disease of classic arrhythmogenic right ventricular dysplasia has therapeutic implications for the patient and relatives. PMID- 10836304 TI - Rosai-Dorfman disease and generalized AA amyloidosis: a case report. AB - We report on a patient who, at 31 years of age, was found to suffer from sinus histiocytosis with massive lymphadenopathy (SHML; Rosai-Dorfman disease) with nodal and extranodal involvement as described previously. Five years later the patient presented with nephrotic syndrome caused by a generalized AA amyloidosis, and he subsequently died from pulmonary thromboembolism owing to renal vein thrombosis. Retrospective analysis of serum levels of C-reactive protein (CRP) showed that during the last 3 years before his death, he had a persistently elevated CRP level ranging from 73 to 161 mg/L, despite antiinflammatory treatment with prednisolone, methotrexate, or 6-mercaptopurine. These figures indicate that the patient was probably suffering from a permanent acute phase response which, in the absence of any other evidence of a chronic inflammatory disease which commonly causes AA amyloidosis, was most likely owing to SHML. PMID- 10836305 TI - Composite lymphoma. PMID- 10836306 TI - Endarterectomy for symptomatic carotid stenosis: some of the lessons learned so far. PMID- 10836307 TI - Type 2 diabetes and tight blood pressure control: how low to go? PMID- 10836308 TI - Third sector primary health care in New Zealand. AB - AIMS: To describe key organisational characteristics of selected third sector (non-profit and non-government) primary health care organisations. METHODS: Data were collected, in 1997 and 1998, from 15 third sector primary care organisations that were members of a network of third sector primary care providers, Health Care Aotearoa (HCA). Data were collected by face-to-face interviews of managers and key informants using a semi-structured interview schedule, and from practice computer information systems. RESULTS: Overall the populations served were young: only 4% of patients were aged 65 years or older, and the ethnicity profile was highly atypical, with 21.8% European, 36% Maori, 22.7% Pacific Island, 12% other, and 7.5% not stated. Community services card holding rates were higher than recorded in other studies, and registered patients tended to live in highly deprived areas. HCA organisations had high patient to doctor ratios, in general over 2000:1, and there were significant differences in management structures between HCA practices and more traditional general practice. CONCLUSIONS: Third sector organisations provide services for populations that are disadvantaged in many respects. It is likely that New Zealand will continue to develop a diverse range of primary care organisational arrangements. Effort is now required to measure quality and effectiveness of services provided by different primary care organisations serving comparable populations. PMID- 10836309 TI - Nosocomial blood stream infection in Auckland Healthcare hospitals. AB - AIM: To report the epidemiology of nosocomial bloodstream infections in Auckland Healthcare Hospitals. METHODS: From January 1995 to December 1997 every positive blood culture result was followed up by an infection control nurse who recorded relevant clinical, laboratory and treatment information on a data collection sheet. The clinical significance of each isolate was determined and the most likely source recorded. RESULTS: During the three year study period, there were 1,046 nosocomial blood stream infections yielding 1,147 isolates. The most common isolates/groups were: coagulase negative staphylococci 19%, S. aureus 18%, E. coli 12%, streptococci 10%, other Enterobacteriaceae 10%, Enterobacter spp. 7%, Pseudomonas spp. 5%, anaerobes 2%, and yeasts 4%. The most common sources were: intravascular lines 40%, urinary tract 8%, skin/soft tissue 8%, gastrointestinal 7%, and unknown 25%. The overall results were strongly influenced by the neonatal intensive care unit at National Women's Hospital where 58% of blood stream infections had intravascular-lines as the source and 53% of the isolates were coagulase negative staphylococci. The overall blood stream infection rate was approximately 6/1,000 admissions. Rates per 1,000 inpatient days for haematology, intensive care, oncology, neonatal and all other patients were 13, 11, 3, 3 and 1 respectively. CONCLUSIONS: Surveillance data that are clinically relevant are useful in identifying areas where infection prevention strategies can be implemented. Because of the importance of lines as a source of nosocomial blood stream infections all aspects of line care are being reviewed with the aim of reducing these devices as a source of blood stream infection. PMID- 10836310 TI - Cumulative incidence of hepatitis C seroconversion in a cohort of seronegative injecting drug users. AB - AIM: To measure the cumulative incidence of hepatitis C virus seroconversion over a two year period in a group of seronegative injecting drug users. METHODS: The study involved follow-up, in 1996, of a cohort (n=85) of injecting drug users identified as hepatitis C virus seronegative in 1994. Participants were interviewed about risk factors for hepatitis C. A blood sample was also taken for anti-hepatitis C virus antibody and hepatitis C virus RNA testing. RESULTS: Forty four participants were interviewed and 39 gave blood for testing. Most (80%) were aged 29 years or under and two thirds (n=26) were male. Around half reported borrowing (49%) or lending (57%) needles and syringes since 1994 and both of these behaviours were associated with seroconversion. The majority (88%) also reported sharing other injecting equipment. Nine were anti-hepatitis C virus positive giving a seroconversion rate over two years of 23% (13 per 100 person years). Four out of the nine seropositive specimens tested were also hepatitis C virus RNA positive. CONCLUSIONS: This study demonstrates a high rate of recent hepatitis C virus seroconversion amongst a group of New Zealand injecting drug users. Transmission of hepatitis C virus appears to be unabated by current control measures. These findings confirm the need to develop more effective policy and practices to prevent further spread, not just of hepatitis C, but of other blood-borne viruses in injecting drug user populations. PMID- 10836311 TI - Intellectual origins and principles of the internal market in New Zealand. PMID- 10836312 TI - Room for more women in clinical specialties. Women in Academic Medicine Working Party. PMID- 10836313 TI - Informed consent. PMID- 10836314 TI - Genetic and environment in asthma: the answer of twin studies. PMID- 10836315 TI - Monitoring airway inflammation in asthma by induced sputum. PMID- 10836316 TI - Genetic and environmental influence on asthma: a population-based study of 11,688 Danish twin pairs. AB - The aim of this study was to estimate the relative influence of genetic and environmental factors on the aetiology of asthma. The classic twin study design was used to analyse data on self-reported asthma obtained by a questionnaire mailed to 34,076 individuals, aged 12-41 yrs and originating from the Danish Twin Register. The cumulative incidence of asthma was 6.2% in the pooled sample. The data showed a decreasing cumulative incidence with age for males, but no sex or zygosity difference was observed. Substantially higher concordance rates, odds ratios and correlations for asthma were estimated in the monozygotic than in the dizygotic twins. Using biometric modelling, a model including additive genetic and nonshared environmental effects provided the best overall fit to the data. According to this model, 73% of the variation in liability to asthma was explained by genetic factors. No sex difference or age-dependency in the magnitude of genetic effects was observed. The biometric analysis emphasized a major influence of genetic factors in the aetiology of asthma. However, a substantial part of the variation in liability to asthma is due to the impact of environmental factors specific to the individual. There is no evidence for a substantial impact of genetic dominance or the shared environment. PMID- 10836317 TI - Prednisone-dependent asthma: inflammatory indices in induced sputum. AB - The kinetics of changes in inflammatory indices in induced sputum from eight prednisone dependent asthmatics whose minimum clinical maintenance and exacerbation doses were known were investigated. The study began on the last day of a course of 30 mg prednisone daily for one week. Thereafter, the daily prednisone was reduced in a structured way to below the maintenance dose. This treatment was continued until a clinical exacerbation occurred. Prednisone 30 mg daily was then given again for one week. The mean duration of prednisone reduction was 7.4 weeks and the median dose was 7.5 mg x day(-1). Increases in sputum eosinophils preceded increases in blood eosinophils by 4 weeks and worsening of symptoms and forced expiratory volume in one second by 6 weeks. The clinical exacerbation was also accompanied by sputum neutrophilia and increases in sputum eosinophil cationic protein (ECP), fibrinogen and interleukin (IL)-5. Treatment with prednisone suppressed median sputum eosinophilia (from 16.3 to 0%, p<0.001), decreased sputum ECP (from 7,480 to 700 microg x L(-1), p = 0.01), but did not improve neutrophil numbers, fibrinogen or IL-5. The results show that the reduction of prednisone treatment in prednisone-dependent asthmatics evokes a severe airway eosinophilic inflammatory response. Clinical and blood indices deteriorate later than those in sputum suggesting that sputum examination may be useful to identify the minimum regular dose of prednisone required in these patients. PMID- 10836318 TI - Antioxidant intake and adult-onset wheeze: a case-control study. Aberdeen WHEASE Study Group. AB - An increase in prevalence of wheezing illness in the UK has coincided with a reduction in the consumption of natural antioxidants, which may modulate the lung's response to oxidant stress, limiting the expression of airway inflammation and respiratory symptoms. The hypothesis that intakes and plasma levels of natural antioxidants would be determinants of adult-onset wheezing illness was tested. A nested case-control study was conducted in 94 cases with adult-onset wheeze and 203 controls aged 39-45 yrs identified in a 30-yr follow-up survey. Antioxidant intake was measured by a food frequency questionnaire, and plasma and red cell measurements of antioxidant status were obtained. Outcome measures were onset of wheeze since age 15 yrs (ever wheeze) and wheeze occurring in the past 12 months (current wheeze). After adjusting for the effects of smoking, socioeconomic status, atopy, family history of atopic disease and total energy intake, intakes of vitamin E (odds ratio (OR) = 4.02 for low compared to high tertile of intake) and plasma levels of ascorbate (OR = 0.98 per unit) and alpha tocopherol:triglyceride ratio (OR = 0.34 per log(e) unit) were inversely related to adult-onset wheeze. In analyses stratified by social class and smoking, intakes of vitamin C and E and plasma levels of ascorbate and alpha tocopherol:triglyceride ratio were inversely related to current wheeze in the manual social class and among current smokers. No independent associations of vitamin A, beta-carotene or total plasma antioxidant capacity were found. The results support the hypothesis that deficiencies of vitamins C and E are associated with wheezing symptoms. Smokers in the manual social class are particularly susceptible to these effects. PMID- 10836319 TI - Lung status in young Danish rurals: the effect of farming exposure on asthma-like symptoms and lung function. AB - The aim of this study was to assess the prevalence of asthma (self-reported) and relate this to lung function and factors associated with asthma in young farmers. Two hundred and ten female and 1,691 male farming students together with 407 males controls were studied. Each subject underwent a medical interview; forced expiratory volume in one second (FEV1) and forced vital capacity (FVC) were recorded using a dry wedge spirometer. Histamine bronchial reactivity was measured using the Yan method. Skin prick testing was performed using inhalant allergens. Nonsmokers had lower prevalence of asthma (5.4-10.8%) than smokers (11.3-21.0%) (p<0.05). Females reported symptoms of asthma nearly twice as often as males. Sex, smoking and a family history of asthma/allergy were significantly associated with asthma. Controls had higher standardized FEV1 and FVC residuals than male students, both nonsmokers (0.21 and 0.24) versus (-0.06 and -0.05) and smokers (0.29 and 0.33) versus (-0.11 and 0.13) (p<0.032). Bronchial hyperresponsiveness, asthma, siblings with allergy and working with cattle (controls only) were significantly associated with reduced lung function. In conclusion, the prevalence of asthma was significantly related to smoking, female sex, family history of asthma and allergy. Whilst bronchial hyperresponsiveness was associated with reduced lung function and lung function was slightly reduced in the male farming students, there was no association found between occupational farming exposure and either lung symptoms or lung function. PMID- 10836320 TI - Reduced intracellular magnesium concentrations in asthmatic patients. AB - Magnesium is important in the regulation of bronchomotor tone, and low dietary intake of magnesium has been associated with airway hyperresponsiveness in epidemiological studies. The concentration of magnesium in serum, erythrocytes and urine in 49 patients with asthma (29 males, aged 15-65 yrs) and in 25 normal subjects (15 males, aged 17-36 yrs) was studied by atomic absorption. Magnesium concentrations were significantly lower in erythrocytes and urine in both atopic (n = 26) and nonatopic (n = 23) asthmatic patients as compared with the control group, whereas serum concentrations did not differ. The concentration of magnesium in erythrocytes was not related to the degree of airway obstruction as measured by forced expiratory volume in one second (FEV1) but was significantly correlated with airway hyperresponsiveness measured as the provocative concentration causing a 20% fall in FEV1 to inhaled acetylcholine (r = 0.64; p<0.05). In addition, a magnesium tolerance test showed increased retention of magnesium (58.9% of administered dose in asthmatic patients compared with 8.9% in normal subjects, p<0.05). In conclusion, the low cellular concentration of magnesium may be associated with airway hyperresponsiveness in asthmatic patients. PMID- 10836321 TI - Allergen challenge-induced acute exudation of IL-8, ECP and alpha2-macroglobulin in human rhinovirus-induced common colds. AB - Rhinovirus infections cause exacerbations of eosinophilic airway disease. The acute effects of allergen-challenge on nasal interleukin-8 (IL-8), eosinophil cationic protein (ECP), and alpha2-macroglobulin were examined in atopic subjects with common cold symptoms. Twenty-three patients with seasonal allergic rhinitis were inoculated with human rhinovirus 16 outside the pollen season. Diluent and allergen challenges, followed by nasal lavages, were carried out about 3 months before and 4 days after virus inoculation. Seventeen patients developed significant common cold symptoms with increased nasal lavage fluid levels of alpha2-macroglobulin, IL-8, and ECP at baseline (p<0.001-0.05 versus before inoculation), and were further increased by allergen challenge (p< 0.001-0.05); IL-8 and ECP levels were correlated (r = 0.63, p<0.001). Before inoculation, the six patients who later did not develop common cold symptoms had high levels of IL 8 and myeloperoxidase (MPO), and exhibited strong allergen-induced plasma exudation responses (alpha2-macroglobulin). After inoculation, IL-8 and ECP did not increase in these symptomless subjects. In conclusion, high nasal interleukin 8 and myeloperoxidase levels and exudative hyperresponsiveness may protect against infection. The association between nasal interleukin-8 and eosinophil cationic protein in common cold, particularly that observed in nasal lavage fluids after allergen-induced acute exudation of plasma, suggests the involvement of interleukin-8 in exacerbation of airway mucosal eosinophil activity. PMID- 10836322 TI - Changes in exhaled carbon monoxide and nitric oxide levels following allergen challenge in patients with asthma. AB - Carbon monoxide is a product of haem degradation by haem oxygenase (HO), activated by inflammatory cytokines and oxidants. This study examined whether allergen challenge can increase exhaled CO levels, as a reflection of HO activation. Exhaled CO and nitric oxide, an expired gas also thought to reflect cytokine-induced airway inflammation, were measured in 15 atopic steroid-naive nonsmoking patients with asthma (13 males, aged 30+/-2 yrs) before and for up to 20 h after allergen challenge. Baseline CO (4.4+/-0.3 parts per million (ppm)) and NO (20.6+/-1.2 parts per billion (ppb)) levels were elevated in asthmatic as compared with nonsmoking normal volunteers (n = 37, 2.1+/-0.2 ppm and 7.0+/-0.1 ppb, respectively, p<0.05). In 10 patients with a dual response in the forced expiratory volume in one second (FEV1) there was a maximal increase in exhaled CO at 1 h (343+/-7.1%) and at 6 h (69+/-12%, p<0.01), followed by a maximal fall in FEV1 (28+/-9%, p<0.05) at 9 h, whereas the maximal NO increase was observed at 10 h (50.2+/-11.8%). The maximal increase in exhaled CO in single response patients (n = 5) was 30+/-2% during the early asthmatic reaction and 46.3+/-9.2% between 4 and 10 h, followed by a fall in FEV1 (9+/-3%, p>0.05) at 9 h, whereas exhaled NO was not significantly changed. In five patients exhaled CO was not attenuated by inhalation of increasing concentrations of histamine causing a 20% fall in FEV1 (PC20) or its subsequent relief by beta2-agonists. In conclusion, exhaled carbon monoxide is increased during the early and late asthmatic reactions independently of the change in airway calibre, while exhaled nitric oxide is increased only during the late reaction and follows the increase in carbon monoxide and fall in the forced expiratory volume in one second in time. PMID- 10836323 TI - Theophylline induces a reduction in circulating interleukin-4 and interleukin-5 in atopic asthmatics. AB - Theophylline, a known phosphodiesterase inhibitor, has been widely used as an additional bronchodilator in asthmatic patients who are not adequately controlled on high-doses of inhaled steroids. However, there is growing evidence that theophylline may also have anti-inflammatory or immunomodulatory effects in asthma. This study investigated whether theophylline administration has an impact on serum levels of interleukin (IL)-4 and IL-5 in asthmatic patients. Eight asymptomatic patients aged 30+/-1.5 yrs (mean +/- SEM) with mild atopic asthma were given a single daily dose of theophylline 150 mg or placebo in an on (theophylline)-off (placebo)-on (theophylline)-off (placebo) protocol with a 3 week duration of each on- or off- interval. Determination of serum IL-4 and IL-5 was done at baseline for all subjects and on the last day of each 3-week interval for the patients under study. Serum IL-4 levels were: 35+/-6 (baseline), 19+/-3 (on-1 interval), 29.5+/-4 (off-2), 15+/-2 (on-3) and 26+/-4 pg x mL(-1) (off-4), while IL-5 levels were 27+/-5, 18+/-4, 28+/-5, 17+/-4 and 28+/-5 pg x mL(-1), respectively. Spirometry was unchanged during the study and serum theophylline levels at the end of the two on-periods were 4.5+/-0.05 and 4.2+/-0.07 microg x mL(-1), while all patients remained asymptomatic. In conclusion, the administration of a low, single, daily dose of oral theophylline in asymptomatic patients with mild atopic asthma seems to reduce circulating interleukin-4 and interleukin-5. PMID- 10836324 TI - Bronchoalveolar lavage findings in patients with chronic nonproductive cough. AB - Mast cells and eosinophils may play a role in the pathophysiology of chronic cough in nonasthmatics. It is unknown, however, whether degranulation of these cells occurs in the airways of such patients. Thirty-five nonsmoking patients referred with a chronic nonproductive cough (mean cough duration 76.2 months) were evaluated using a comprehensive diagnostic protocol. Bronchoalveolar lavage (BAL) cell differentials and BAL histamine, tryptase and eosinophilic cationic protein (ECP) concentrations were determined. Ten nonsmoking healthy volunteers served as controls. Diagnostic subgroups were identified: eight postnasal drip syndrome (PNDS), seven cough variant asthma (CVA), seven gastro-esophageal reflux (GOR), seven dual aetiology and six idiopathic. Nonasthmatic coughers (NAC) were characterized as those patients without bronchial hyperresponsiveness on histamine challenge and whose cough had either responded to therapy for PNDS or GOR or failed to improve with antiasthma therapy. There was a significant increase in both eosinophil and mast cell numbers (p<0.05) and in histamine levels (p = 0.027) when NAC patients were compared with controls. Tryptase and ECP levels were elevated in 7 of 23 and 6 of 23 NAC patients, respectively. In conclusion, airway inflammatory cell numbers are not only increased but also activated, suggesting an important role for airways inflammation in the pathophysiology of chronic nonproductive cough. PMID- 10836325 TI - Increased expression of activation markers and adhesion molecules on lung T-cells compared with blood in the normal rat. AB - Lymphocytes play an important role in many lung diseases and are routinely accessible by bronchoalveolar lavage (BAL). Lymphocytes from the BAL (BAL pool) have a different subset composition to those from peripheral blood, consisting mainly of activated T-cells. The aim of this study was to examine whether preferential migration of activated T-cells to the bronchoalveolar space or factors of the specific microenvironment mediate this phenomenon. The expression of adhesion molecules and cellular activation markers (intercellular adhesion molecule-1, leukocyte function-associated antigen-1, CD2, CD44, interleukin-2 receptor and L-selectin) was studied on T- and B-cells not only in the BAL and peripheral blood (blood pool), but also in the compartments in between, such as the lung vascular perfusate (marginal pool) and the lung interstitium (interstitial pool), with the experiments being performed simultaneously in the same animals. Low levels of adhesion molecule expression were observed on T-cells in the blood and marginal pool, medium levels in the lung interstitium and the highest levels in the BAL. "Memory" (CD45R(low)) and "naive" (CD45R(high)) T cells in the lung compartments showed a higher expression of adhesion molecules compared with blood. However, the predominating CD45R(low) T-cells showed a significantly higher expression than the CD45R(high) cells, indicating that CD4+ CD45R(high) T-cells had changed their phenotype to CD45R(low). In conclusion, a high level of expression of leukocyte function associated antigen-1 and intracellular adhesion molecule-1 on the bronchoalveolar lavage and interstitial T-cells is more likely to be the result of local, lung-specific induction than a prerequisite for migration into the bronchoalveolar space. PMID- 10836326 TI - ICAM-1 mediates lung leukocyte recruitment but not pulmonary fibrosis in a murine model of bleomycin-induced lung injury. AB - Bleomycin-induced lung injury has been extensively used as a model of interstitial pneumonia and pulmonary fibrosis. Intercellular adhesion molecule (ICAM)-1 is a ligand for lymphocyte function-associated antigen (LFA)-1alpha and has been shown to be required for leukocyte migration into inflamed areas. The purpose of this report was to investigate the role of the ICAM-1/LFA-1alpha pathway in a murine model of bleomycin-induced lung injury. Animals received 75 mg x kg(-1) bleomycin (BLM) i.v. followed by treatment with phosphate-buffered saline (BLM group), anti-ICAM-1 and LFA-1alpha monoclonal antibodies (mAb) (BLM+mAb group). Inflammatory cell counts of bronchoalveolar lavage (BAL) fluid, hydroxyproline content and histological findings were compared between these groups. In the BLM group, significant increases in total cell count, macrophage count and neutrophil count of BAL fluid were observed on days 7 and 14. In the BLM+mAb group, bleomycin-induced accumulation of neutrophils was significantly reduced on days 7 and 14 (p<0.01). However, the administration of mAb to ICAM-1 and LFA-1alpha did not decrease the lung hydroxyproline content or the histopathological fibrosis grading score, indicating that the antagonism of ICAM I and LFA-1alpha did not attenuate bleomycin-induced pulmonary fibrosis. This study suggests that the intercellular adhesion molecule-1/lymphocyte function associated antigen-1alpha pathway mediates the accumulation of inflammatory cells in the injured lung caused by bleomycin; however, other mechanisms are important for the subsequent development of pulmonary fibrosis. PMID- 10836327 TI - The bronchoprotective effect of inhaled salmeterol in preschool children: a dose ranging study. AB - The optimal dose of salmeterol in infants and preschool children is not known. The aim of this study was to assess the bronchoprotective effect of different doses of salmeterol using methacholine-induced wheeze in children aged <4 yrs. Children <4 yrs old with a history of recurrent wheeze underwent two methacholine challenges within 7 days. One hour before each challenge they were pretreated in double-blind fashion using a metered-dose inhaler and Babyhaler spacer. Placebo was given before one challenge, and either 25, 50 or 100 microg of salmeterol before the other. Both the dose and treatment order were random. The provocative concentration of methacholine causing wheeze (PCwheeze) was measured on each occasion. Studies were terminated when wheeze occurred or arterial oxygen saturation (Sa,o2) fell below 91%. Of the 42 children enrolled, 33 completed the study. Two subjects refused the challenge test, two failed to return and five developed upper respiratory tract infections or wheeze between the two tests. The mean (range) age of the population was 27 (8-46) months. Ratios of PCwheeze between treatment and placebo challenges were calculated for each dosage group. The treatment/placebo ratios (95% confidence intervals) were 1.2 (0.6-2.4) for 25 microg, 2.5 (1.4-4.6) for 50 microg (p<0.01), and 4.0 (2.1-7.4) (p<0.001) for 100 microg doses. In recurrently wheezy children aged <4 yrs a single dose of salmeterol between 25 and 100 microg has a dose-dependent effect on methacholine induced wheeze, and this is significantly different from placebo at 50 and 100 microg. This study suggests that the Babyhaler effectively delivers salmeterol to children <4 yrs of age and that doses between 50-100 microg are efficacious. PMID- 10836328 TI - Lung function in school-aged asthmatic children with inhaled cromoglycate, nedocromil and corticosteroid therapy. AB - Two-thirds of the children with asthma in our area use cromones and only one third steroids as the maintenance therapy. This study aimed to evaluate our treatment policy based on the international consensus. Peak expiratory flow (PEF), dynamic spirometry and bronchodilation test results were therefore collected in 195 school-aged patients who visited our outpatient clinic in 1995. Sixty-four children (33%) used cromoglycate, 86 (44%) nedocromil and 45 (23%) inhaled steroids. Twenty-five (12%) needed combination therapy, mainly with salmeterol. Lung function results were good, and there were no significant differences between the therapeutic groups irrespective of whether pre- or postbronchodilator values were considered. PEF was decreased in eight (4%), forced expiratory volume in one second (FEVI) in four (2%) and maximum mid expiratory flow (MMEF) in 33 (17%) patients. At least one result was decreased in 39 (20%) cases, in most cases (77%) MMEF alone. Significant rises after salbutamol inhalations were observed in 17 (9%) in PEF, in two (1%) in FEV1 and 20 (10%) in MMEF values. Thus, the bronchodilation test was positive in 33 (17%) cases, and in 22 (11%) cases it was the only sign of bronchial obstruction. Over 70% of the children with asthma can be treated with cromones by a stepwise treatment modality. Inhaled steroids can be restricted to those not controllable by cromones. Lung function tests, including postbronchodilator values, should be part of the follow-up of continuous maintenance medication for asthma. PMID- 10836329 TI - Systemic effects of inhaled corticosteroids on growth and bone turnover in childhood asthma: a comparison of fluticasone with beclomethasone. AB - Inhaled steroids are frequently used in childhood asthma, but concerns based on limited objective evidence remain, regarding long-term side-effects. In this study the systemic effects of standard doses of inhaled steroids in childhood asthma were assessed, comparing beclomethasone dipropionate (BDP) with fluticasone propionate (FP). The study was prospective, randomized and double blind. Twenty-three steroid-naive children with moderately severe asthma, aged 5 10 yrs, were allocated either BDP (400 microg x day(-1) or FP (200 microg x day( 1)) using a metered-dose inhaler with a spacer. Asthma control was assessed at regular intervals over 20 months. Fasting morning blood and overnight urine samples were collected for estimation of serum cortisol, serum 1-carboxyterminal telopeptide (ICTP), serum osteocalcin and urine deoxypyridinoline (DPD). Bone mineral density (BMD) was measured at each visit. None of the markers of bone turnover showed any change during the study period. BMD increased at normal rates with age. Serum cortisol significantly decreased on BDP, but not on FP. A significant difference in growth rates was found between the groups, with a slower rate of growth towards the end of the observation period in the BDP group. In conclusion when taken in a relatively modest dose over a period of time, beclomethasone dipropionate had significant effects on the hypothalamic-pituitary adrenal axis and statural growth in childhood asthma. These systemic effects were not seen with an equipotent dose of fluticasone propionate. PMID- 10836330 TI - Seasonality, but not prevalence of sudden infant death syndrome varies by region in mainland Britain. AB - This study aimed to investigate whether seasonal variation in day length contributed to winter/summer variation in sudden infant death syndrome (SIDS) at different latitudes in mainland Britain. Over 11 yrs 13,973 deaths were studied. Using appropriate analytic techniques a sine curve was fitted to monthly rates with the amplitude indicating magnitude of seasonal change. The rate of SIDS per 1,000 live births was the same (1.73) in the north as in the south. The amplitude was a quarter less in the north (41.3%) than in the south (54.2%) (p<0.001). While annual rates did not differ, the within year distribution did. The findings for seasonality of SIDS births were similar (amplitudes: north 213%, south 32.3%). Correlations were made between SIDS amplitude and individual environmental factors, particularly temperature and day length. These complex issues, while reported briefly, do not allow firm conclusions. In the north the winter day length is shorter, sunshine hours are less and temperature is lower, but the winter increment in SIDS is less. The extent of seasonal variation of sudden infant death syndrome is greater in the south as compared with the colder, darker north but this has no effect on sudden infant death syndrome rates. Changing photoperiod by latitude, amongst other environmental influences, may hold clues to the aetiology of sudden infant death syndrome. PMID- 10836331 TI - Genotype-phenotype correlations in cystic fibrosis: clinical severity of mutation S549R(T-->G). AB - With a view to assessing genotype-to-phenotype correlations in cystic fibrosis (CF), the clinical presentation of CF children from the United Arab Emirates (UAE) who were homozygous for cystic fibrosis transmembrane conductance regulator (CFTR) mutation S549R(T-->G was investigated. This mutation is localized in intron 11 (nucleotide binding domain 1 of the CFTR protein) and had so far been described as a private mutation only. The associations between the R549/R549 genotype and 20 outcome variables, including age at diagnosis, sweat chloride concentrations, growth percentiles, meconium ileus, pancreatic sufficiency, pulmonary disease, associated complications and micro-organism colonization were examined in a group of 15 CF children (9 females and 6 males). Mean current age and age at diagnosis were both low (5.4+/-3.5 and 1.0+/-1.1 yrs, respectively). Although none of the 15 CF patients had presented with meconium ileus at birth, all were pancreatic insufficient and had very severe lung disease, with a high rate of Pseudomonas aeruginosa and Staphylococcus aureus. Two patients died during the course of this investigation (one was 5 months and the other, 6 yrs old). The clinical presentation associated with S549R(T-->G) homozygosity in the United Arab Emirates is quite homogeneous and shows an extreme degree and course of cystic fibrosis severity. PMID- 10836332 TI - Expired hydrogen peroxide in breath condensate of cystic fibrosis patients. AB - Stimulated inflammatory cells release large amounts of hydrogen peroxide (H2O2). Breath condensate H2O2 has been shown to be elevated in stable asthmatic children, chronic obstructive pulmonary disease and intubated adult respiratory distress syndrome. In cystic fibrosis airways, where neutrophilic inflammation dominates, it is postulated that H2O2 in breath condensate would be elevated and may be used as a marker of airways inflammation. Expired breath condensate was collected from 16 clinically stable cystic fibrosis (CF) patients (mean age 25.3 yrs, mean forced expiratory volume in one second (FEV1) 50.2%) and 14 normal subjects (mean age 29.9 yrs). Total plasma leukocyte, neutrophil, monocyte and eosinophil counts and lung function were also measured on the day of collection. A method of breath condensate collection excluding the confounding factors of nasal air and saliva contamination was validated and used and H2O2 measured fluorometrically using an optimized assay. The median level of H2O2 concentration in breath condensate of CF patients was lower than that in normal subjects (0.064 versus 0.089 microM), but this did not reach statistical significance (p = 0.20, Mann-Whitney rank sum test). Within the CF group, there was no correlation between H2O2 concentration and lung function. Expired breath condensate H2O2 is not elevated in patients with cystic fibrosis, and is thus not a suitable marker of airways inflammation in these patients. Possible explanations include physical barriers to its detection caused by viscous airways secretions, reaction with other reactive species or increased antioxidant activity caused by trapping of positively charged antioxidants in negatively charged airways secretions. PMID- 10836333 TI - Recombinant DNase in cystic fibrosis: a protocol for targeted introduction through n-of-1 trials. Scottish Cystic Fibrosis Group. AB - Nebulized recombinant human deoxyribonuclease (DNase) reduces sputum viscosity and improves lung function in some cystic fibrosis patients, but individual responses are unpredictable. The aim of this study was to investigate how DNase can be targeted to those cystic fibrosis patients who would benefit most. The Scottish Cystic Fibrosis Group agreed on a randomized, double-blind, placebo controlled n-of-1 assessment protocol. Patients underwent a maximum of three 4 week assessment periods (2 weeks saline, 2 weeks DNase each). Measurements performed at hospital (exercise, oximetry and spirometry) and home (symptom scores) were used to derive a scoring system to discriminate maximally between DNase and placebo effects. The data on 89 4-week assessments in 52 patients were reported. Twenty-four patients have completed the assessment process (12 responders and 12 nonresponders) to date. Forced expiratory volume in one second (FEVI) was the best discriminator of response, rising by >200 mL after DNase in 33 of 89 (37%) assessments compared with 3 of 89 (3%) after saline. N-of-1 trials, while laborious, permitted genuine treatment effects to be quantified within individuals with confidence, permitting appropriate treatment targeting. This provides a model of how other new expensive therapies may be introduced to maximize patient benefit. PMID- 10836334 TI - Effect of L-arginine infusion on airway NO in cystic fibrosis and primary ciliary dyskinesia syndrome. AB - Airway nitric oxide concentrations in patients with cystic fibrosis or primary ciliary dyskinesia syndrome have been shown to be lower than in healthy subjects. Decreased NO concentrations may contribute to impaired ciliary clearance, respiratory tract infections, or obstructive lung disease in these conditions. Nasal and exhaled NO concentrations were compared before and after infusion of 500 mg x kg(-1) L-arginine, the substrate of NO synthases, in 11 cystic fibrosis (CF) patients, seven primary ciliary dyskinesia (PCD) syndrome patients, and 11 control subjects. Baseline nasal and exhaled NO concentrations were significantly lower in both CF and PCD syndrome patients than in controls (p<0.01). In controls, the maximum increase of NO was seen immediately after L-arginine infusion in the upper airways (1.8-fold) and 3 h after the infusion in the lower airways (1.4-fold). Although NO concentrations also increased significantly in both CF (1.9-fold and 1.6-fold, respectively) and PCD syndrome patients (1.4-fold and 1.8-fold, respectively), concentrations remained subnormal compared with baseline values of controls. Pulmonary function remained unchanged in both patient groups. In conclusion, the low airway nitric oxide formation in both cystic fibrosis and primary ciliary dyskinesia syndrome patients can be augmented by L-arginine administration. The finding that pulmonary function remained unchanged in both conditions may be due to the fact that normalization of airway nitric oxide concentrations could not be achieved. PMID- 10836335 TI - Effects of oxygen on autonomic nervous system dysfunction in patients with chronic obstructive pulmonary disease. AB - Chronic hypoxaemia can play a pathological role in abnormalities of the autonomic nervous system (ANS). In patients with chronic obstructive pulmonary disease (COPD), chronic hypoxaemia is associated with increased mortality and only long term oxygen therapy is able to improve their survival. Normoxaemic COPD patients have been shown to suffer from abnormalities in ANS function. The aims of this study were to evaluate ANS function in COPD patients with chronic hypercapnic respiratory insufficiency and to test whether oxygen supplementation could reverse any ANS dysfunction. Eleven stable COPD patients with chronic hypercapnic respiratory insufficiency underwent evaluation of ANS by analysis of variability in cardiac frequency at rest and during both vagal (controlled breathing) and sympathetic (tilting) stimuli breathing with and without oxygen supplementation. Thirteen male, healthy, nonsmoking volunteers served as controls. Evaluation of ANS in COPD patients during hypoxic conditions showed alterations both at rest and in response to vagal and sympathetic stimuli. Oxygen supply reversed hypoxaemia without significant changes in arterial carbon dioxide tension and, therefore, ANS alterations were corrected during sympathetic stimulus only. Breathing room air and oxygen, the resting low-frequency (LF) powers were 45+/-15 and 148+/-55 ms2 x Hz(-1), respectively, and controlled breathing LF were 107+/ 41 and 141+/-113 ms2 x Hz(-1), respectively. In stable patients with chronic obstructive pulmonary disease with chronic respiratory insufficiency, hypoxaemia is associated with derangements in the autonomic nervous system which may be partially reversed by oxygen administration. PMID- 10836336 TI - Long-term effectiveness of pulmonary rehabilitation in patients with chronic airway obstruction. AB - The aim of this study was to evaluate the long-term outcome of an outpatient pulmonary rehabilitation programme (PRP) in patients with chronic airway obstruction (CAO). In 61 CAO patients (35 asthmatics and 26 chronic obstructive pulmonary disease (COPD)) lung and respiratory muscle function, exercise tolerance (by symptom limited cycloergometer and walking tests), dyspnoea (Borg scale, visual analogue scale (VAS), baseline and transitional dyspnoea index (BDI and TDI, respectively)) and quality of life (St George's Respiratory Questionnaire (SGRQ)) were assessed at baseline (to), at discharge (t1) and 12 months postdischarge (t2). Preprogramme and post-programme hospital admissions and exacerbations of disease were also recorded. In comparison with baseline, no significant change was observed in lung function tests in either diagnostic group, either at t1 or at t2. In both groups improvements in respiratory muscle strength, exercise tolerance, Borg scale and VAS reported at t1 were partially reduced at t2. Analysis of variance showed that these changes over time were similar in the two groups. Mean values of SGRQ and BDI/TDI improved at t1, and, unlike exercise tolerance, did not worsen at t2. However, a clinically relevant difference in SGRQ between t2 and to was reported only in 56% of asthmatics and 52% of COPD patients. Compared with the preceding 2 yrs, in the year following PRP, hospital admissions and disease exacerbations decreased significantly in both diagnostic groups. Regardless of diagnosis, patients with chronic airway obstruction who underwent an outpatient pulmonary rehabilitation programme maintained an improved quality of life 12 months postdischarge despite a partial loss of the improvement in exercise tolerance. PMID- 10836337 TI - Clearance of inhaled technetium-99m-DTPA as a clinical index of pulmonary vascular disease in systemic sclerosis. AB - This study evaluated the utility of the clearance time of inhaled diethylenetriamine pentaacetate (DTPA) to distinguish pulmonary vascular disease from early fibrosing alveolitis (FA) in patients with systemic sclerosis (SSc). It was hypothesized that this would be preserved in patients with vascular disease compared with FA, despite similar gas-transfer deficits and matching lung volumes, because of the preservation of alveolar epithelial integrity. All patients had SSc and were categorized into a control group (C; n=9), pulmonary vascular group (VAS; n=14) or FA group (n=14) dependent on the appearance on a computed tomography (CT) scan and the transfer factor of the lung for carbon monoxide (T(L,CO)) (VAS and FA < or =70%, C > or =80%). All patients had a forced vital capacity (FVC) of >80%. The T(L,CO) (median) was similar in the VAS (57.5%) and FA (60%) groups. There was a significant difference in median DTPA clearance half-times between FA (21.25 min) and VAS (46.5 min) (p=0.014) and between FA and C (84.5 min) (p=0.0004). No difference was found between VAS and C (p=0.0778). Follow-up data from the VAS group showed no subsequent development of FA on the CT scan and no decrease in FVC (n=13, mean 42 months). These results suggest that clearance of diethylenetriamine pentaacetate is preserved in patients likely to have pulmonary vascular disease and may be useful in distinguishing fibrosing alveolitis from vascular disease in systemic sclerosis. PMID- 10836338 TI - An imbalance in plasma prostanoids in patients with Raynaud's phenomenon and pulmonary vasospasm. AB - Raynaud's phenomenon has been suggested as a predisposing factor for pulmonary vasospasm which may lead to pulmonary hypertension, but the occurrence of cold stimulus-induced pulmonary vasospasm has been inconsistent. Such inconsistent pulmonary vascular responses may be caused by differences in the production of endogenous vasodilators and vasoconstrictors among patients. Fourteen patients with Raynaud's phenomenon associated with mixed connective tissue disease (n=10) or systemic sclerosis (n=4) participated in the study. Right heart catheterization was performed before and after a cold pressor test, immersing a hand in cold water (15 degrees C) for 5 min. Plasma levels of 6-keto prostaglandin (PG)F1alpha, thromboxane (TX)B2 and endothelin (ET)-1 in the mixed venous blood were measured. Mean pulmonary artery pressure increased after the cold pressor test in five of 14 patients, and the patients were divided into those with pulmonary vasospasm (responders) and those without vasospasm (nonresponders). After the cold pressor test, levels of 6-keto PGF1alpha increased significantly in nonresponders (p<0.01) and decreased significantly in responders (p<0.05). The ratios of 6-keto PGF1alpha to TXB2 significantly increased in nonresponders (p<0.01) but not in responders and the difference between responders and nonresponders after the cold pressor test was also statistically significant (p<0.05). No significant change in plasma ET-1 levels occurred in either responders or nonresponders. The results suggest that an impaired production of prostaglandin I2 and an imbalance between prostaglandin I2 and thromboxane A2 are associated with the occurrence of pulmonary vasospasm induced by Raynaud's phenomenon. PMID- 10836339 TI - Endothelin(A)-receptor antagonism attenuates pulmonary hypertension in porcine endotoxin shock. AB - Porcine endotoxin shock is characterized by pulmonary hypertension, decreased mean arterial pressure and deteriorated cardiac performance. These pathophysiological findings are accompanied by increased plasma endothelin-1 levels. Previous studies have shown that both the pulmonary and systemic circulation are improved by combined endothelinA- and endothelinB-receptor antagonism. This study was designed to evaluate further the specific involvement of the endothelinA-receptor in cardiopulmonary pathophysiology during endotoxin shock. In a porcine endotoxin shock model, the endothelinA-receptor antagonist PD 155080 was administered after the onset of endotoxaemia. Cardiopulmonary vascular changes, dynamic lung compliance, oxygen-related variables and plasma levels of endothelin-1-like immunoreactivity were compared with a control group receiving only endotoxin. PD 155080 counteracted the increase in pulmonary artery pressure seen in control animals. In contrast, cardiac performance was not improved. Dynamic lung compliance was not affected by PD 155080. Plasma levels of endothelin-1-like immunoreactivity increased to a similar degree in both groups. These findings indicate that the endothelin system is involved in the pathophysiology of endotoxin shock. Furthermore, the change in pulmonary circulation seen in the present shock model is to a large extent mediated by the endothelinA-receptor mechanism. In view of previous findings, the deterioration of cardiac performance may involve the endothelinB-receptor, either alone or in combination with the endothelinA-receptor. PMID- 10836340 TI - The efficacy of a custom-fabricated nasal mask on gas exchange during nasal intermittent positive pressure ventilation. AB - Commercially available nasal masks have a large mask volume and give rise to considerable air leaks around the mask during nasal intermittent positive pressure ventilation (NIPPV) which may reduce alveolar ventilation (VA per breath). The effects of a custom-fabricated nasal mask (F-mask) versus a commercially available mask (C-mask) on arterial blood gas measurements, dead space including both physiological and apparatus dead space (VD), air leak and VA per breath were compared in patients with restrictive thoracic disease during short-term NIPPV sessions while using a volume cycled ventilator with equivalent settings for both masks. The mask volume of the C-mask was significantly larger than that of the F-mask (p<0.003). The arterial carbon dioxide tension (Pa,CO2) during NIPPV with either the F-mask (5.56+/-1.35 kPa) (mean+/-SD) or the C-mask (6.87+/-0.96 kPa) was significantly lower than during spontaneous breathing (7.75+/-0.81 kPa; p<0.003), but the Pa,CO2 decreased more during NIPPV with the F mask than with the C-mask (p<0.003). The VD was significantly smaller (p<0.03), the air leak was significantly less (p<0.03), and the VA per breath was significantly larger (p<0.03) during NIPPV with the F-mask than with the C-mask. In conclusion, nasal intermittent positive pressure ventilation with the F-mask was more effective than nasal intermittent positive pressure ventilation with the commercially available mask due to its smaller dead space and less air leak. Further studies are needed to extend these results to all the commercially available-masks. PMID- 10836341 TI - Mechanisms underlying effects of nocturnal ventilation on daytime blood gases in neuromuscular diseases. AB - The hypothesis that, in neuromuscular and chest wall diseases, improvement in central respiratory drive explains the effects of night-time ventilation on diurnal gas exchanges was tested. The effects at 6 months, 1, 2 and 3 yrs of intermittent positive pressure ventilation (IPPV) on arterial blood gas tension, pulmonary function, muscle strength, sleep parameters, respiratory parameters during sleep and ventilatory response to CO2 were evaluated in 16 consecutive patients with neuromuscular or chest wall disorders. As compared with baseline, after IPPV daytime arterial oxygen tension (Pa,O2) increased (+2.3 kPa at peak effect) and arterial carbon dioxide tension (Pa,CO2) and total bicarbonate decreased (-1.8 kPa and -5 mmol x L(-1), respectively) significantly; vital capacity, total lung capacity, maximal inspiratory and expiratory pressures and alveolar-arterial oxygen gradient did not change; the apnoea-hypo-opnoea index and the time spent with an arterial oxygen saturation (Sa,O2) value <90% decreased (-24 and -101 min, respectively), sleep efficiency and mean Sa,O2 increased (+16% and +5%, respectively); and ventilatory response to CO2 increased (+4.56 L x min(-1) x kPa(-1)) significantly. The reduction in Pa,CO2 observed after IPPV correlated solely with the increase in the slope of ventilatory response to the CO2 curve (r=-0.68, p=0.008). In neuromuscular or chest wall diseases, improvement of daytime hypoventilation with nocturnal intermittent positive pressure ventilation may represent an adaptation of the central chemoreceptors to the reduction of profound hypercapnia during sleep or reflect change in the quality of sleep. PMID- 10836342 TI - Respiratory and haemodynamic effects of the prone position at two different levels of PEEP in a canine acute lung injury model. AB - This study was designed to examine whether the oxygenation response in the prone position differs in magnitude depending on the level of positive end-expiratory pressure (PEEP) applied in the supine position, and whether cardiac output (CO) increases in the prone position. In seven supine dogs, acute lung injury was established by saline lavage (arterial oxygen tension (Pa,O2)/inspiratory oxygen fraction (FI,O2) 17.8+/-9.6 kPa (134+/-72 mmHg)), and inflection point (Pflex) of the respiratory system was measured (6.6+/-1.4 cmH2O). Pa,O2/FI,O2 and CO of the supine and prone positions were obtained under the application of low PEEP and then under optimal PEEP (2 cmH2O below and above Pflex, respectively). The net increase in Pa,O2/FI,O2 by prone positioning was greater at low PEEP (27.3+/-12.0 kPa (205+/-90 mmHg)) than at optimal PEEP (4.4+/-13.0 kPa (33+/-98 mmHg)) (p=0.006). CO decreased significantly with optimal PEEP in the supine position (2.4+/-0.5 versus 3.1+/-0.4 L x min(-1) at baseline, p<0.001), and increased to 3.4+/-0.6 and 3.6+/-0.7 L x min(-1) in the prone position at 5 min and 30 min, respectively (both p=0.018). When the dogs were turned supine at optimal PEEP, CO again decreased (2.4+/-0.5 L x min(-1), p<0.001). In conclusion, the prone position augmented the effect of relatively low positive end-expiratory pressure on oxygenation, and attenuated the haemodynamic impairment of relatively high positive end-expiratory pressure in a canine acute lung injury model. PMID- 10836344 TI - Search for agents causing atypical pneumonia in HIV-positive patients by inhibitor-controlled PCR assays. AB - Pneumonia is one of the most frequent complications in acquired immunodeficiency syndrome-patients with Pneumocystis carinii as the leading cause. The true prevalence of atypical agents such as Chlamydia pneumoniae, C. trachomatis, Legionella pneumophila and Mycoplasma pneumoniae in this population of patients is unknown as the currently used method for diagnosing these agents is measurement of antibody levels. However, this method is of limited value in human immunodeficiency virus (HIV)-positive patients who may have a compromised antibody response. To evaluate the prevalence of Chlamydia spp., Legionella spp. and M. pneumoniae in HIV-infected patients with pulmonary disease, this retrospective study has applied inhibitor-controlled polymerase chain reaction analyses on 103 bronchoalveolar lavage (BAL) fluids representing 103 episodes of pneumonia in 83 HIV-positive patients. L. pneumophila was detected in 1% of the BAL fluids and M. pneumoniae was found as a coexisting pathogen in 2% of the samples. Chlamydia spp. could not be detected in any of the BAL fluids. By culture and staining methods 106 other microorganisms were detected with P. carinii and Streptococcus pneumoniae as the most frequently occurring. Pneumonia due to Chlamydia pneumoniae, Legionella pneumophila or Mycoplasma pneumoniae seems to be rare in Danish human immunodeficiency virus-infected patients, but might be considered as a possible cause in cases of treatment failure. PMID- 10836343 TI - Oxidatively modified proteins in bronchoalveolar lavage fluid of patients with ARDS and patients at-risk for ARDS. AB - Oxidative stress in acute respiratory distress syndrome (ARDS) is considered as an important pathophysiological mechanism in acute impairment of lung function. The present study investigated whether a pulmonary oxidant-antioxidant imbalance is indicated by substantial oxidative modification of proteins in bronchoalveolar lavage (BAL) fluid. Oxidatively modified proteins in BAL fluid, as measured by the reduction of protein carbonyl groups with tritiated borohydride, were studied in control subjects, patients with clinically established ARDS, and patients considered at-risk for ARDS because they had had coronary bypass surgery. Subsets of these at-risk patients were pretreated either with methylprednisolone or N acetylcysteine. The carbonyl content of BAL fluid proteins was greatly increased in ARDS patients (5.0+/-13 nmol carbonyl x mL(-1) BAL fluid; mean+/-SEM; p=0.0004; n=10) and moderately increased in the untreated patients at-risk for ARDS (1.3+/-0.2 nmol x mL(-1); p=0.027; n=19) compared with controls (0.8+/-0.2 nmol x mL(-1); n=12). The two other at-risk groups pretreated either with methylprednisolone or N-acetylcysteine showed carbonyl values that were statistically not different from the controls (1.2+/-0.2 nmol x mL(-1); p=0.13; n=13, and 1.1+/-0.3 nmol x mL(-1); p=0.40; n=8, respectively). These results show that oxidatively modified proteins clearly accumulated in bronchoalveolar lavage fluid of acute respiratory distress syndrome patients, and to a minor extent in untreated at-risk patients. These data suggest a severe oxidant-antioxidant imbalance in acute respiratory distress syndrome. PMID- 10836345 TI - Pre-emptive therapy with azoles in lung transplant patients. Geneva Lung Transplantation Group. AB - Pulmonary fungal infection is diagnosed in up to 15-25% of lung transplant recipients and frequently bears a fatal outcome. This prospective uncontrolled study addresses the efficacy and safety of pre-emptive azole therapy against fungal infection in these patients. Fluconazole or itraconazole have been systematically used according to reported fungus sensitivity after the discovery of fungi in lower respiratory tract samples. Patients were treated until the bronchial suture was normal and the cultures of the following bronchoscopy remained negative. Fungi were found post-transplantation in the lower respiratory tract specimens of 26 out of 31 (84%) patients, predominantly Candida albicans (20 patients) and Aspergillus fumigatus (16 patients). Mycelia characteristic of Candida spp. or Aspergillus spp. were found in necrotic tissue at the bronchial suture of nine patients. The mean duration of the 38 treatments was 3.6+/-2.6 months (range, 0.5-12 months). After a median follow-up of 16 (range, 0-48) months, two cases of extended ulcerative and pseudo membranous Aspergillus fumigatus bronchitis were observed and healed under itraconazole treatment. In conclusion, pre-emptive azole therapy may be effective and well-tolerated in lung transplant patients where fungi are found in the airways or pleura. PMID- 10836346 TI - Health-based selection for asthma, but not for chronic bronchitis, in pig farmers: an evidence-based hypothesis. AB - In cross-sectional surveys among pig farmers an elevated prevalence of chronic bronchitis is usually reported, but not of asthma, despite many factors promoting the manifestation of asthma. To investigate this apparent contradiction a postal questionnaire survey was performed among 239 pig farmers and 311 rural controls. Pig farmers reported an elevated prevalence of symptoms of chronic bronchitis (20.2 versus 7.7%, p<0.001), but not of asthma (5.9 versus 5.5% for chest tightness). Among pig farmers the use of disinfectants (prevalence odds ratio (POR) 9.4, 95% confidence interval (CI) 1.6-57.2 for quaternary ammonium compounds) and aspects of the disinfecting procedure were associated with the prevalence of asthma symptoms. Compared with controls, pig farmers reported fewer allergies to common allergens (4.6 versus 14.6%, p<0.001 for pollen) and fewer symptoms of atopy in childhood (9.9 versus 17.2, p<0.05 for one or more of four symptoms). Atopy in childhood was strongly associated with the prevalence of asthma symptoms (POR 4.1, 95% CI 2.2-7.7), but not with chronic bronchitis. Health-based selection of nonasthmatics for pig farming, which tends to mask a work-related hazard for asthma, is offered as an explanation for these results. PMID- 10836347 TI - Role of neutrophil elastase in hypersecretion in asthma. AB - Goblet cell (GC) hyperplasia and mucous plugging are common in patients with acute asthma. These patients also show neutrophil recruitment into the airways. Neutrophils contain elastase, a potent secretagogue in airways. Therefore, it was reasoned that neutrophil recruitment, by releasing elastase, could result in GC hypersecretion. When neutrophil chemoattractants were instilled in the airways of guinea-pigs, time-dependent neutrophil recruitment and GC degranulation occurred. An inhibitor of leukocyte infiltration (NPC15669) prevented both responses, implicating neutrophils. An inhibitor of neutrophil elastase (ICI 200,355) abolished GC degranulation, implicating elastase. Further studies implicate movement of elastase from cytoplasmic granules to the neutrophil surface, and they suggest a role for adhesion molecules on neutrophils and on GCs in neutrophil-dependent GC degranulation. Similarly, instillation of ovalbumin (OVA) into airways of OVA-sensitized guinea-pigs caused early recruitment of neutrophils and GC degranulation. GC degranulation was prevented by pretreatment with NPC15669 or ICI 200,355. These results implicate neutrophil release of elastase in allergen-induced hypersecretion. The results suggest a mechanism for the mucous plugging that occurs in acute asthma; prevention of neutrophil recruitment, prevention of neutrophil-GC adhesion, or inhibition of elastase activity could provide effective therapy for this serious pathophysiological abnormality. PMID- 10836348 TI - Physiological changes in respiratory function associated with ageing. AB - Physiological ageing of the lung is associated with dilatation of alveoli, enlargement of airspaces, decrease in exchange surface area and loss of supporting tissue for peripheral airways ("senile emphysema"), changes resulting in decreased static elastic recoil of the lung and increased residual volume and functional residual capacity. Compliance of the chest wall diminishes, thereby increasing work of breathing when compared with younger subjects. Respiratory muscle strength also decreases with ageing, and is strongly correlated with nutritional status and cardiac index. Expiratory flow rates decrease with a characteristic alteration in the flow-volume curve suggesting small airway disease. The ventilation-perfusion ratio (V'A/Q') heterogeneity increases, with low V'A/Q' zones appearing as a result of premature closing of dependent airways. Carbon monoxide transfer decreases with age, reflecting mainly a loss of surface area. In spite of these changes, the respiratory system remains capable of maintaining adequate gas exchange at rest and during exertion during the entire lifespan, with only a slight decrease in arterial oxygen tension, and no significant change in arterial carbon dioxide tension. Ageing tends to diminish the reserve of the respiratory system in cases of acute disease. Decreased sensitivity of respiratory centres to hypoxia or hypercapnia results in a diminished ventilatory response in cases of heart failure, infection or aggravated airway obstruction. Furthermore, decreased perception bronchoconstriction and diminished physical activity may result in lesser awareness of the disease and delayed diagnosis. PMID- 10836349 TI - A breathless female. PMID- 10836350 TI - Isosorbide-5-mononitrate in the treatment of pulmonary hypertension associated with portal hypertension. AB - Pulmonary hypertension is uncommonly associated with portal hypertension. The current approach for the management of pulmonary hypertension involves the use of vasodilators in patients who show vascular responsiveness during an acute challenge. Since the association of portal hypertension with pulmonary hypertension is very seldomly presented, its optimal therapy has not been defined. Moreover, calcium-channel blockers, which are usually used in pulmonary hypertension treatment, may exert a deleterious effect on portal hypertension. Therefore, the search for drugs that may be active under both conditions has important clinical implications. This report presents the case of a patient with portal hypertension-associated pulmonary artery hypertension that was effectively treated with isosorbide-5-mononitrate (Is-5-Mn). The patient had severe portal hypertension (hepatic venous pressure gradient=14.5 mmHg) and pulmonary hypertension (mean pulmonary artery pressure (PAP)=50 mmHg). Acute administration of prostacyclin and nitric oxide elicited a significant reduction in both PAP and pulmonary vascular resistance (PVR), an effect that was also achieved with Is-5 Mn. The patient was treated with 40 mg Is-5-Mn twice daily and a haemodynamic study performed 6 months later showed that the reduction in both PAP and PVR persisted. PMID- 10836351 TI - Occupational asthma caused by palladium. AB - Occupational exposure to complex platinum salts is a well-known cause of occupational asthma. Although there is evidence that platinum refinery workers may also be sensitized to other precious metals, such as palladium or rhodium, no instances of occupational asthma due to an isolated sensitization to palladium have been reported. A case is reported of occupational rhinoconjunctivitis and asthma in a previously healthy worker exposed to the fumes of an electroplating bath containing palladium. There was no exposure to platinum. Sensitization to palladium was documented by skin-prick tests. The skin-prick test was positive with Pd(NH3)4Cl2, but not with (NH4)2PdCl4. Corresponding salts of platinum were all negative. A bronchial provocation test with Pd(NH3)4Cl2 (0.0001% for a total of 315 s, followed by 0.001% for a total of 210 s) led to an early decrease in forced expiratory volume in one second (-35%). A similar exposure (0.001% for a total of 16 min) in an unrelated asthmatic gave no reaction. This case shows that an isolated sensitization to palladium can occur and that respiratory exposure to palladium is a novel cause of metal-induced occupational asthma. PMID- 10836352 TI - Activation of lymphocytes and increased interleukin-5 levels in bronchoalveolar lavage fluid in acute eosinophilic pneumonia. AB - Acute eosinophilic pneumonia (AEP) is a recently described illness and the number of case reports has increased during the last few years. However, the role of interleukin (IL)-5 and activated lymphocytes in the pathogenesis or activity of AEP is still not clear. The clinical features, lymphocyte surface analysis and IL 5 concentrations in bronchoalveolar lavage fluid (BALF) and peripheral blood (PB) of a young female patient with AEP are described before and at 2 weeks, 4 weeks and 6 months after a 3-day course of i.v. methylprednisolone. Serum and BALF concentrations of IL-5 before treatment were 5,200 and 8,400 pg x mL(-1), respectively. Activated CD4 lymphocytes bearing CD25 and human leukocyte antigen (HLA)-DR in BALF were higher than in PB. Treatment caused a rapid fall in these cells and levels of IL-5 in BALF returned to normal levels in parallel with clinical improvement. There was no evidence of recurrence after cessation of steroid therapy. In contrast, eosinophilia in BALF persisted for 4 weeks after steroid therapy in spite of normalization of the chest radiograph and arterial blood gases. The number of CD8+CD11b- (suppressor/cytotoxic) T-cells subsequently increased while the number of CD8+CD11b+ cells decreased. These results suggest that activated CD4 cells and interleukin-5 elevation contribute to the development of acute eosinophilic pneumonia rather than persistent eosinophilia in the lung and that a short course of steroid therapy may effectively control acute eosinophilic pneumonia. PMID- 10836353 TI - Asthma mortality: summary of a round-table discussion, New York, January 1997. PMID- 10836354 TI - Endothelin-1 expression in airway epithelial cells. PMID- 10836355 TI - Is there an association between bronchial asthma, food allergy/intolerance and analgesic intolerance? PMID- 10836356 TI - Ethical issues in increasing living kidney donations by expanding kidney paired exchange programs. AB - INTRODUCTION: In 1997, Ross et al. proposed to increase the supply of living kidney donations by using kidneys from living ABO-incompatible donors through an exchange arrangement between two living kidney donor-recipient pairs. Although many transplant centers are exploring this option, only a small fraction of potential donor-recipient pairs are eligible for an exchange on the basis of ABO incompatibility. In this article, we explore three variations that have potentially great clinical relevance. METHODS: The three potential variations discussed are: (1) altruistically unbalanced living donor-recipient exchanges; (2) an indirect exchange (an exchange between a living donor-recipient pair with a cadaveric donor-recipient pair) on the basis of a positive crossmatch; and (3) an indirect exchange on the basis of ABO incompatibility. DISCUSSION: The goal of kidney paired exchange programs is to increase the supply of kidneys available for transplantation ethically. We acknowledge that all exchanges increase the potential for coercion, and we currently reject the proposal of altruistically unbalanced exchanges because of the potential for coercion. However, we believe that voluntary consent can be achieved for indirect exchanges. The indirect ABO compatible exchange creates no new ethical concerns to our original living paired exchange program and we support its implementation. The indirect ABO-incompatible exchange does create a new ethical concern because it may increase the vulnerability of O blood group recipients. If mechanisms can be developed to avoid increasing the waiting time for blood group O recipients, we would support the implementation of the indirect ABO-incompatible exchange. PMID- 10836357 TI - The management of cyclosporine-induced nephrotoxicity after cardiac transplantation. PMID- 10836358 TI - Another way of presenting alloantigens to the immune system? PMID- 10836359 TI - Genetic predisposition to thrombosis in renal transplant recipients: the factor V Q506 (Leiden) allele. PMID- 10836360 TI - Clinical transplantation tolerance twelve years after prospective withdrawal of immunosuppressive drugs: studies of chimerism and anti-donor reactivity. AB - BACKGROUND: Previous studies showed the feasibility of inducing transplantation tolerance to cadaveric renal allografts in patients given pretransplant total lymphoid irradiation (TLI). Microchimerism has been theorized to be an important or necessary factor in long-term graft acceptance and tolerance in humans. METHODS: A cadaveric renal transplant recipient given pretransplant total lymphoid irradiation and withdrawn from immunosuppressive drugs more than 12 years ago was tested for microchimerism using a sensitive nested polymerase chain reaction technique, and for anti-donor reactivity using the mixed leukocyte reaction and an ELISA screen for anti-HLA antibodies. Donor and recipient were mismatched for all HLA-A, B, and DR antigens. RESULTS: The "tolerant" recipient had good graft function, no detectable donor-type cells in the blood by polymerase chain reaction analysis, vigorous reactivity to donor stimulator cells in the mixed leukocyte reaction, and no detectable serum anti-HLA antibodies. CONCLUSIONS: Operational tolerance to HLA-A, B, and DR mismatched organ allografts can be induced prospectively in humans for at least 12 years after withdrawal of immunosuppressive drugs. The allograft can be maintained in the absence of detectable donor microchimerism and in the presence of anti-donor reactivity in the mixed leukocyte reaction, suggesting that neither chimerism nor clonal deletion or anergy of recipient T cells to alloantigens presented by donor Class II HLA molecules is required for persistence of the tolerant state using this total lymphoid irradiation protocol. PMID- 10836361 TI - Compromised kidney graft rejection response in Vervet monkeys after withdrawal of immunosuppressants tacrolimus and sirolimus. AB - BACKGROUND: In nonprimates, organ allografts are often not rejected after withdrawal of immunosuppression. In this study, we examined whether such a phenomenon also occurs in primates. METHODS: Vervet monkeys were transplanted with renal allografts and treated for 60 days with tacrolimus, or tacrolimus plus sirolimus. The drugs were totally withdrawn on day 61. The survival of the monkeys was monitored, and their response to donor- or third party-derived alloantigens was examined in vivo and in vitro. RESULTS: The majority (80-100%) of the grafts survived for at least additional 30 days with no signs of acute rejection. The compromised rejection is donor-specific, because recipient monkeys failed to reject a donor-derived skin graft, but a third-party skin graft was rejected. In vitro mixed lymphocyte reaction and interleukin-2 production in the mixed lymphocyte reaction between the recipients and their donors or between the recipients and a third party had no discernable patterns, and thus did not reflect the in vivo status of the immune system. Although the recipients could not reject the graft acutely after drug withdrawal, the kidney grafts and the donor-derived skin grafts had pathological findings of chronic rejection. CONCLUSIONS: The rejection response of the monkeys to an established graft after withdrawal of immunosuppression is compromised. The compromised rejection is specific and is not due to a permanent alteration of the immune system by the initial drug treatment. The allografts are not inert but have low levels of interaction with the recipient immune system. PMID- 10836362 TI - Transplantation of H-2Kb-transgenic adrenocortical cells in the mouse having undergone an adrenalectomy: functional and morphological aspects. AB - BACKGROUND: A new model of cellular adrenocortical transplantation after bilateral adrenalectomy in the mouse was established. This model was used to study the effects of the expression of the transgenic MHC class I molecule H 2K(b) (Kb) on graft survival and morphologic features, corticosterone secretion, and the possibility of tolerance induction in the recipient. METHOD: A single cell suspension of purified adrenocortical cells was grafted under the kidney capsule of B10.Br (H-2k) mice having adrenalectomies. Syngeneic, fully MHC mismatched, and MHC class I-incompatible Kb-transgenic mice served as donor strains. To analyze graft function, urinary excretion and serum levels of corticosterone were monitored over 100 days. Tolerance induction in the graft recipients of Kb-transgenic and third party skin grafts was tested on day 50 after adrenocortical transplantation. Histological sections of the adrenocortical grafts were obtained on day 100. RESULTS: Recipients of syngeneic and Kb transgenic grafts displayed pretransplant corticosterone levels on days 20, 50, and 100 and ACTH-stimulated serum corticosterone levels similar to those of controls on day 100 after adrenocortical transplantation. In contrast, in recipients of fully MHC-mismatched grafts, corticosterone excretion was significantly reduced. In this group, 4 of 7 mice did not survive. Syngeneic skin grafts survived indefinitely in recipients of syngeneic and Kb-transgenic adrenocortical grafts, whereas Kb-transgenic and fully MHC-mismatched skin grafts were acutely rejected. Tissue sections of the adrenocortical grafts revealed vascularized cell conglomerates in syngeneic and Kb-transgenic grafts without infiltrations of mononuclear cells. Furthermore, a differentiation similar to adrenocortical organization was partly found. CONCLUSION: In conclusion, a model of cellular adrenocortical transplantation was established. The results show that syngeneic transplantation resulted in physiological corticosterone levels early after transplantation, whereas fully MHC-incompatible grafts were rejected. Recipients of Kb-transgenic grafts showed unimpaired adrenocortical function, but did not tolerize toward Kb-transgenic skin grafts. Possible mechanisms include a local immunomodulatory effect of glucocorticoids secreted by the graft and a low immunogenicity of the relatively small numbers of transplanted cells. PMID- 10836363 TI - Induction of unresponsiveness to islet xenograft by MMC treatment of graft and blockage of LFA-1/ICAM-1 pathway. AB - BACKGROUND: Induction of unresponsiveness to graft is one of major interest in xenotransplantation. Two different modalities [direct graft treatment by mitomycin C (MMC) and blockage of the lymphocyte function-associated antigen 1/intracellular adhesion molecule-1 (LFA-1/ICAM-1) pathway in recipients by species-specific mAbs] were tested for their ability to produce unresponsiveness to secondary islet xenografts. METHODS: Collagenase-digested WS (RT1k) rat islets, purified by Ficoll density gradient, were incubated for 30 min with MMC 10 microg/ml, cultured for 20 hr, and transplanted into the renal subcapsular space of streptozotocin-induced diabetic C57BL/6 (H-2b) mice. Recipient mice were divided into experimental groups according to anti-rat ICAM-1 and/or anti-mouse LFA-1 mAb treatment and transplantation of MMC-treated or nontreated islets. RESULTS: MMC pretreatment alone prolonged graft survival, with a mean survival time (MST) of 23.0+/-7.4 days, compared with that of cultured islets (12.4+/-2.7 days; P<0.01). MMC treatment of islets significantly augmented graft survival, compared with that of crude islet grafts under treatment with anti-donor ICAM-1 mAb (MST: >41.3+/-30 vs. 16.6+/-5.4 days, P<0.01), anti-recipient LFA-mAb (MST: >70.3+/-28.9 vs. 30.4+/-10.4 days, P<0.001), or both mAbs (MST: >88.1+/-24.1 vs. 23+/-7.4 days, P<0.0001). One of six, four of nine, and six of eight animals accepted MMC-treated islet xenografts over 100 days after treatment with anti-rat ICAM-1, anti-mouse LFA-1, or both mAbs treatments, respectively, whereas none of the animals accepted nontreated islets under the same treatment. When the mice bearing long-term functioning xenografts were challenged with the secondary graft from the original donor strain, the animals previously treated with anti recipient LFA-1 and anti-donor ICAM-1 mAbs were more prone to accept it than animals given anti-recipient LFA-1 mAb alone (MST: 55.8+/-25.7 vs. 15+/-2.4 respectively; P<0.001), although they rejected the third-party xenograft and allograft acutely. CONCLUSIONS: In the xenogeneic islet transplantation model, MMC graft pretreatment and blockage of the ICAM-1/LFA-1 pathway constitute a potent protocol for inducing unresponsiveness to islet xenografts. PMID- 10836364 TI - Altered distribution and density of myocardial beta-adrenoceptors during acute rejection in rats. AB - BACKGROUND: A better understanding of altered expression of myocardial beta adrenoceptors can facilitate the diagnosis of early and late acute rejection of heart transplants. METHODS: We submitted rats to heterotopic heart transplantation (iso- and allografts) of which one of each were treated with or without cyclosporin A for 4, 7, and 14 days (n=5, respectively). The cardiac sections were incubated in vitro with [3H]CGP 12177, or the hearts were labeled in vivo by intravenous injection of [3H]CGP 12177. Autoradiographic images of both were analyzed digitally and compared with histologic findings. RESULTS: Beta adrenoceptor distribution in native hearts and isografts was homogeneous, but highly differential distribution and density in allografts were observed in the left and right ventricular walls and in the interventricular septum despite treatment with cyclosporin A. High-density areas in the progressive course of acute rejection are commonly associated with up-regulation of beta-adrenoceptors in apparently viable myocytes, although histologic findings confirmed many infiltrating mononuclear cells. Low-density areas, which were identified in the right and left ventricular walls as early as 4 days after transplantation, correlated with derangement of myocytes, which was suggestive of early acute rejection. The images obtained by in vivo technique were comparable to in vitro images. CONCLUSIONS: The alteration of beta-adrenoceptor expression in allografts showed a close relationship with the severity of acute rejection, and the techniques employed in this model were useful in our study of the rejection process and in detecting early and late acute rejection in the rat. PMID- 10836365 TI - Factor V R506Q mutation (activated protein C resistance) is an additional risk factor for early renal graft loss associated with acute vascular rejection. AB - BACKGROUND: The factor V R506Q mutation (FV R506Q, FV:Q506, or FV Leiden) resulting in activated protein C (APC) resistance is the most common inherited risk factor for venous thrombosis, including in renal transplant recipients. We investigated a possible association between the FV mutation and early renal graft loss, and the prevalence of macro- and microvascular thrombosis, endothelialitis, and fibrinoid vascular necrosis by FV genotype. METHOD: One hundred and nine renal allograft recipients were genotyped for FV mutation. A vascular rejection subgroup of patients (n=29) had experienced at least one episode of vascular rejection, or graft thrombosis. A second group of patients (n=80) had experienced no acute rejection and retained a well-functioning graft. RESULTS: The prevalence of APC resistance was numerically but not statistically significantly higher in the vascular rejection group (17.2%) compared with the group without rejection episodes (7.5%) (P=0.16). There was a significant association between the presence or absence of FV mutation and graft survival, with a 55.6% 1-year graft survival rate versus a 76.4% rate, respectively (P=0.02). The prevalence of vascular rejection, as evidenced by endothelialitis or fibrinoid vascular necrosis, was significantly associated with APC resistance but macro- or microvascular thrombosis were not. CONCLUSION: Renal transplant recipients who are carriers of the FV:Q506 allele have an increased risk of early graft loss. Vascular rejection changes including endothelialitis and fibrinoid vascular necrosis were more common in this group, and therefore an association between the hypercoagulable state, which entails an up-regulation of the mitogenic and proinflammatory enzyme thrombin, and the immunological challenge to the endothelium may be the cause of inferior prognosis in these patients. PMID- 10836366 TI - Serum immunoglobulin levels in relation to levels of specific antibodies in allogeneic and autologous bone marrow transplant recipients. AB - BACKGROUND: The aim of this study was to investigate the correlation of total levels of immunoglobulins to levels of specific antibodies after allogeneic and autologous bone marrow transplantation. Autologous transplant patients had normal levels of IgA and IgG antibodies already at 6 months after transplantation. In allogeneic transplanted patients without chronic graft versus host disease the immunological recovery was slower. The IgA and IgG levels were at the limit for deficiency at 6 months after transplantation. In allogeneic transplant patients with chronic chronic graft versus host disease the immunological recovery was delayed further. The total IgG levels were low at 12 months after transplantation and the IgG subclass pattern did not normalize until 24 months after transplantation. IgA levels remained low at 24 months after transplantation in all allogeneic transplanted patients with chronic chronic graft versus host disease. Protective levels of specific antibodies against tetanus and pneumococci decreased during the first year after transplantation regardless of the total immunoglobulin levels, regardless of the donors immunity. Pneumococcal antibodies decreased only in allogeneic transplanted patients, although autologous transplant patients retained pretransplant immunity against pneumococci. There was no difference in levels of specific antibodies between patients with and without chronic chronic graft versus host disease at 12 months after transplantation. There was no correlation between total immunoglobulin levels to levels of specific antibodies against tetanus and pneumococci after transplantation in our study. Taken together, normalized immunoglobulin levels do not predict normalization of levels of specific antibodies against tetanus and pneumococci after transplantation. PMID- 10836367 TI - Improvement of impaired renal function in heart transplant recipients treated with mycophenolate mofetil and low-dose cyclosporine. AB - BACKGROUND: Cyclosporine (CsA) nephrotoxicity is a common problem after cardiac transplantation. We have studied the impact of CsA dose reduction in association with mycophenolate mofetil (MMF) treatment on renal function in heart transplant recipients with suspected CsA nephrotoxicity (serum creatinine level >2 mg/dl). METHODS: Twelve heart transplant recipients (11 men, 1 woman; 111 to 1813 days after transplantation) with CsA-based immunosuppression (plus azathioprine and/or steroids) and a serum creatinine level >2.0 mg/dl were started on a daily dose of 2000 mg of MMF. Dilated cardiomyopathy was the underlying disease in nine patients, ischemic cardiomyopathy in three patients. Mean patient age was 57 years (range 44-69 years). Azathioprine was discontinued and CsA slowly tapered. Creatinine clearance, serum creatinine level, urea nitrogen, and uric acid were monitored. CsA levels were measured, and CsA dose was adjusted for whole blood levels of 70-120 microg/L. Ten patients still had endomyocardial biopsies, whereas one had echocardiographic controls only. RESULTS: One grade 1B rejection episode according to ISHLT (International Society for Heart and Lung Transplantation) was observed until 1 year after the switch to MMF. One patient was excluded due to gastrointestinal side effects. CONCLUSIONS: Conversion from azathioprine to MMF with consecutive reduction of CsA in heart transplant recipients with CsA-impaired renal function improves renal function as evidenced by lower serum creatinine, urea nitrogen, uric acid, and higher creatinine clearance. PMID- 10836368 TI - Relation between steroid dose, body composition and physical activity in renal transplant patients. AB - BACKGROUND: Fat mass is increased in renal transplant (RTx) patients, which may have untoward metabolic and cardiovascular effects. The influence of steroids on body composition (BC), resting energy expenditure (REE), and substrate oxidation rates was assessed in stable RTx patients in a cross-sectional design. Also, the relation between physical activity and nutrient intake, respectively, and body composition was studied. METHODS: 77 RTx patients (42 males, 35 females) were studied. Twenty-one patients were on 10 mg and 27 patients on 5 mg maintenance steroid dose; 29 patients were receiving steroid-free immunosuppression. Assessed were BC (DEXA, anthropometry), REE and substrate oxidation (indirect calorimetry), physical activity (Baecke questionnaire), and nutrient intake (dietary records). RESULTS: BC was not different between the 0-, 5-, and 10-mg steroid group, and no relationship existed between cumulative dose of steroids and BC. REE and substrate oxidation also did not differ between the various groups, apart from a small increase in glucose and decrease in lipid oxidation in female patients using 5-mg steroids. Especially in females, leisure time physical activity was positively related with the percentage lean body mass (r=0.571, P=0.004) and inversely related with fat mass (r= -0.588, P=0.003). Nutrient intake and BC (corrected for physical activity) were not related. CONCLUSIONS: No relation was observed between daily and cumulative steroid dosage and BC and between daily steroid dose and REE and substrate oxidation in RTx patients. Especially in female patients, physical activity level and the percentage of lean body mass concluded and body fat were significantly related. PMID- 10836369 TI - Clinicopathological analysis of liver allograft biopsies with late centrilobular necrosis: a comparative study in 54 patients. AB - BACKGROUND: Centrilobular necrosis (CLN) in liver allografts can be a difficult lesion to interpret histologically. Although long recognized in association with developing chronic rejection, recent studies have described the lesion in association with a number of other disease processes. To clarify the histologic features that could allow a specific diagnosis to be made and to determine the outcome in different diagnostic groups, we assessed biopsies from 54 patients with CLN. METHODS: Biopsies were classified as CLN with acute cellular rejection (ACR), CLN with hepatitis, CLN with developing chronic rejection (CR), and CLN of other etiology. Histologic features were assessed and then compared between groups, and clinical outcomes were noted. RESULTS: Discriminating features for the different groups were as follows: CLN and ACR showed bile duct injury, endothelialitis, and acinar congestion. CLN and CR showed severe bile duct injury, bile duct loss, or centrilobular swelling. CLN and hepatitis was often a diagnosis of exclusion, although interface hepatitis was more common in this group. Cases of autoimmune hepatitis usually demonstrated plasma cell predominance in the portal and acinar inflammatory infiltrate. Significantly, there was considerable overlap in the histologic features between the groups, accounting for the diagnostic difficulty. Patients in whom the CLN was associated with CR or vascular complications generally required retransplantation or died, but in the groups with ACR and hepatitis, the outcome was more favorable. CONCLUSIONS: With regard to most liver allograft biopsies showing late CLN, it is possible to make a specific diagnosis despite overlapping histologic features; this allows specific therapy to be instituted. Ultimately this is likely to contribute to improved graft survival. PMID- 10836370 TI - Cardiac myosin autoantibodies and acute rejection after heart transplantation in patients with dilated cardiomyopathy. AB - OBJECTIVES: To determine whether humoral autoimmune responses associated with dilated cardiomyopathy (DCM) influence the postoperative clinical course following cardiac transplantation. METHODS: ELISA levels of preformed cardiac myosin (CM) autoantibodies (Abs) in patients with a pretransplant diagnosis of dilated cardiomyopathy (DCM) (n=64) and ischemic heart disease (IHD, n=53) were correlated with cardiac rejection, immunosuppression, and the incidence of endocardial infiltrates after transplantation. RESULTS: Alpha- and beta-CM autoantibody (IgG and IgM) levels were similar in DCM and IHD patients but were statistically higher than in controls. Distribution of preformed (beta-CM) IgM Abs in patients with and without rejection in the first postoperative year differed in the two groups. DCM patients rejected earlier P=0.006, and the frequency of rejection at 3 months was statistically higher than in IHD patients. Frequency and reactivity of IgM-Abs in DCM patients with rejection [International Society for Heart and Lung Transplant (ISHLT) grade I and above] was 28% compared with 7% in rejection-free patients, P<0.05. IgM-positive patients had a greater frequency and severity of rejection episodes and required more immunosuppression. These patients had rejection earlier than Ab-negative patients, P<0.009. There was no correlation between antibody status and rejection in IHD patients or with IgG in either group. Distribution of IgG subclass differed in the two diseases. DCM patients had significantly higher IgG3 reactivity; 70% of this activity was present in patients who developed moderate rejection. IgG3-positive patients experienced more frequent rejections, as well as a greater incidence of grade 3A/B rejection as the first episode, than did Ab-negative patients (50% vs. 15%), P<0.05. Frequency of endocardial infiltrates was statistically higher in IgG3 positive patients. CONCLUSION: Proinflammatory characteristics of preformed IgG3 and IgM antibodies in DCM patients may influence the frequency and severity of cardiac rejection after transplantation. PMID- 10836371 TI - Infections in pediatric kidney and liver transplant patients after perioperative hospitalization. AB - BACKGROUND: Infectious complications are a major cause of morbidity and mortality after organ transplantation. There are several reports on infections during the first months after transplantation, but there are very few data regarding infections in long-term survivors of pediatric organ transplantation. METHODS: The incidence and type of infections were retrospectively analyzed in 56 children who underwent 59 liver or renal transplantations. Follow-up was begun when the patient was sent home after a successful operation. All of the children received triple immunosuppression. RESULTS: During a mean follow-up of 4.8 years (total, 286 patient years), 1540 episodes of infection were recorded. The median incidence was 4.8 episodes/patient year. The greatest number was seen in the smallest children, 3 to 6 months after transplantation. Viral upper respiratory tract infections were the most common problem, accounting for half of the episodes (2.7 episodes/patient year). Gastroenteritis was the second most common viral infection. Only 45 episodes of infection with herpesviruses were recorded, and seven of those were caused by cytomegalovirus. Otitis media and sinusitis were the most common bacterial infections and complicated upper respiratory infection in 23% of episodes. Thirty-nine episodes of urinary tract infections were diagnosed, thirty-one in children with renal transplants. Other bacterial infections were rare, and only three episodes of verified bacterial sepsis were diagnosed. CONCLUSION: The frequency and type of infections in children with liver and renal transplants who are on triple immunosuppression are quite similar to those in age-matched healthy children. PMID- 10836372 TI - Long-term renal outcome after lung transplantation is predicted by the 1-month postoperative renal function loss. AB - BACKGROUND: Progressive renal function loss is common after lung transplantation. To facilitate the design of renoprotective strategies, identification of early predictors for long-term renal function loss would be useful. METHODS: We prospectively analyzed renal function [glomerular filtration rate (GFR); 125I iothalamate clearance] in a closely monitored cohort (minimum 24-month follow-up) of 57 patients who received lung transplants between November 1990 and September 1996 in our center. The analyzed end points were the slope of the GFR from 6 months posttransplant onward and the GFR at 24 months after transplantation. RESULTS: Before transplantation, the GFR was 100 ml/min (median, range 59-163). It decreased to 67 ml/min (29-123) at 6 months, 53 ml/min (17-116) at 24 months, and 51 ml/min (20-87) at 36 months after transplantation. The magnitude of the loss of GFR 1 month post-transplantation was the only factor significantly correlated with absolute GFR at 24 months after transplantation. Pulmonary diagnosis was significantly associated with long-term rate of renal function loss. Median loss of GFR was greatest in patients with cystic fibrosis (-10 ml/min/year, range -14 to -6 ml/min/year), preserved in pulmonary hypertension ( 1 ml/min/year, range -6 to +7 ml/min/year), and in between in emphysema (-6 ml/min/year, range -27 to +12 ml/min/year). No other factors could be identified. CONCLUSIONS: In lung transplant recipients, the 1-month postoperative loss of GFR is an early marker for long-term renal prognosis. Pulmonary diagnosis appears to be a relevant predictor as well. These factors may guide further research and the development of preventive strategies. PMID- 10836373 TI - Osteonecrosis after lung transplantation: cystic fibrosis as a potential risk factor. AB - BACKGROUND: Osteonecrosis is a known complication after transplantation of solid organs. The incidence of osteonecrosis after lung transplantation is not well documented. METHODS: We investigated the incidence of symptomatic osteonecrosis in lung transplant recipients, transplanted between November 1992 and June 1998 at our institution. For the detection of osteonecrosis, all patients complaining of musculoskeletal pain underwent magnetic resonance imaging. Demographic characteristics, time after transplantation, etiology of underlying lung disease, and the number of steroid pulses for rejection episodes were compared for patients with and without osteonecrosis. RESULTS: Of 63 transplant recipients, all 49 with a follow-up of >3 months were included for analysis. Of seven symptomatic transplant recipients, five cases of osteonecrosis (10%) were detected at a median duration of 216 days (range 44-600) after transplantation. Patients with osteonecrosis have been treated with the same immunosuppressive regimen and with an equal number of steroid pulses for acute rejection episodes (1.4+/-1.1 vs. 1.4+/-1.5, P=0.69), but were younger (26+/-8 vs. 40+/-11 years, P<0.01) than other transplant recipients. Symptomatic osteonecrosis was detected in four of 14 patients (29%) with cystic fibrosis (CF), compared with one osteonecrosis among 35 patients (3%) with other underlying diseases (P<0.02). Within the group of CF patients, specific clinical and demographic characteristics correlating with the risk for subsequent osteonecrosis could not be found. CONCLUSION: In lung transplant recipients, CF may be a risk factor for the development of symptomatic osteonecrosis. PMID- 10836374 TI - Liver transplantation for cholangiocarcinoma: results in 207 patients. AB - BACKGROUND: Because of the high incidence of recurrent tumor, many surgeons have become disenchanted with transplantation as a treatment for cholangiocarcinoma. METHODS: The Cincinnati Transplant Tumor Registry database was used to examine 207 patients who underwent liver transplantation for otherwise unresectable cholangiocarcinoma or cholangiohepatoma. Specific factors evaluated included tumor size, presence of multiple nodules, evidence of tumor spread at surgery, and treatment with adjuvant chemotherapy and/or radiation therapy. Incidentally found tumors were compared to tumors that were known or suspected to be present before transplantation. RESULTS: The 1, 2, and 5-year survival estimates using life table analysis were 72, 48, and 23%. Fifty-one percent of patients had recurrence of their tumors after transplantation and 84% of recurrences occurred within 2 years of transplantation. Survival after recurrence was rarely more than 1 year. Forty-seven percent of recurrences occurred in the allograft and 30% in the lungs. Tumor recurrence, and evidence of tumor spread at the time of surgery, were negative prognostic variables. There were no positive prognostic variables. Patients with incidentally found cholangiocarcinomas did not have improved survival over patients with known or suspected tumors. A small number of patients survived for more than 5 years without recurrence. However, this group had no variable in common that would aid in the selection of similar patients in the future. CONCLUSIONS: Because of the high rate of recurrent tumor and lack of positive prognostic variables, transplantation should seldom be used as a treatment for cholangiocarcinoma. For transplantation to be a viable treatment in the future, more effective adjuvant therapies are necessary. PMID- 10836375 TI - Activation of alloreactive T cells by allogeneic nonprofessional antigen presenting cells and interleukin-12 from bystander autologous professional antigen-presenting cells. AB - BACKGROUND: After solid organ transplantation most alloantigens are presented to the recipient's immune system by normal tissue cells, which can be considered to act as nonprofessional antigen-presenting cells (APC). It is well accepted that such nonprofessional APC fail to activate recipient resting T cells due to their inability to deliver costimulatory activity. In our study, we tested a hypothesis that such costimulatory activity may be provided by "bystander" recipient professional APC. METHODS: We set up mixed lymphocyte cultures (MLC) of purified T cell responders and T cell stimulator cells, the latter as nonprofessional APC carrying allogeneic MHC class I and II, and tested if responder-type autologous APC could facilitate responder T cell proliferation. In this assay also the effects of anti-CD28 antibody and interleukin- (IL) 1beta, IL-6, or IL-12 mediated costimulation on responder T cell proliferation and IL-2 production were investigated. RESULTS: Autologous APC, i.e., monocytes, were found to facilitate the proliferative response of resting T cells stimulated by allogeneic nonprofessional APC. IL-12 was identified as the most important costimulatory factor for induction of proliferation. IL-1beta enhanced IL-2 production and proliferation of allostimulated resting T cells but its presence was not essential. Although CD28 triggering alone was ineffective, this costimulatory pathway enhanced IL-2 production and proliferation when combined with IL-12 or IL 1beta. CONCLUSIONS: We conclude that costimulatory activity for activation of resting human T cells by nonprofessional donor APC can be delivered through activity of bystander recipient-type autologous APC. This mechanism of allostimulation may contribute to the induction and perpetuation of alloreactivity "in vivo" in a time frame when intragraft professional donor-type APC have been replaced with professional recipient-type APC. PMID- 10836376 TI - Quantitative and qualitative differences in thrombopoietin-dependent hematopoietic progenitor development between cord blood and bone marrow. AB - BACKGROUND: To assess the clinical application of thrombopoietin (TPO) for thrombocytopenia of patients receiving cord blood (CB) or bone marrow (BM) transplants, we examined whether various types of hematopoietic progenitors including megakaryocyte (MK) progenitors from CB and BM exerted different proliferative and differentiative potential in the presence of TPO. METHODS: The development of MK, granulocyte-macrophage, and erythroid/mixed erythroid (E/Mix) progenitors in a serum-deprived liquid culture medium supplemented with TPO was compared between CD34+ CB and BM cells. RESULTS: The CD34+ CB cells generated 30 fold more MKs than the CD34+ BM cells, but the CB-derived MKs were more immature. A single-cell culture study showed that CB CD34+CD38- cells as well as CD34+CD38+ cells proliferated in response to TPO, whereas the two subpopulations of CD34+ BM cells showed little multiplication. In short-term liquid cultures containing CD34+ CB or BM cells, TPO significantly increased the absolute numbers of various types of colony-forming cells, compared with the input values. In particular, MK progenitors and E/Mix progenitors in CB were amplified to a substantially greater extent than in BM. The superior response of CD34+ CB cells to TPO observed in this study may be due in part to the use of cryopreserved cells. CONCLUSIONS: Our results suggest that TPO alone cannot only stimulate megakaryocytopoiesis but also increase the numbers of various types of hematopoietic progenitors, and that quantitative and qualitative differences in TPO-dependent hematopoietic progenitor development exist between CB and BM. PMID- 10836377 TI - Characterization of cytokine profiles in corneal allograft with anti-adhesion therapy. AB - BACKGROUND: The cytokine profiles of specific immunosuppression after anti adhesion molecule therapy are unknown in a mouse corneal transplantation model. METHODS: Orthotopic mouse corneal transplantation was performed using BALB/c (H 2d) mice as recipients and C3H/He (H-2k) mice as donors. Anti-mouse very late antigen-4 and anti-mouse lymphocyte function-associated antigen-1 monoclonal antibodies (each at a dose of 0.25/mg/day) were administered i.p. until day 7. A second corneal transplantation was performed 5 weeks after the first grafting. Delayed hypersensitivity was tested after the second grafting. Corneal cytokine expression was examined immunohistochemically. The cytokine gene transcription level was assessed in the corneas and splenocytes. RESULTS: All allografts with anti-adhesion molecule therapy survived for 5 weeks. Two weeks after the second grafts in the fellow eye (7 weeks after the first grafts), 50% of the mice with successful grafts bilaterally had low delayed hypersensitivity responses. Low helper T 1 (interferon-gamma and interleukin-2) cytokine gene and protein expression in corneas was observed in monoclonal antibody-treated mice 3 weeks after the first grafting. The mice with successful second grafts showed low corneal T helper 1 cytokine gene and protein expression. High interleukin-4 gene transcription levels in corneas and splenocytes was obtained in both groups in which the grafts were accepted and rejected after the second grafts. CONCLUSIONS: The cytokine profile to differentiate alloantigen-specific acceptance with anti adhesion therapy to lymphocyte function-associated antigen-1 and very late antigen-4 molecules from rejection after the second grafting is local and systemic low T helper 1 cytokine in corneal transplantation. High interleukin-4 cytokine expression in corneas and splenocytes is not associated with achievement of tolerance induction. PMID- 10836378 TI - Role of porcine P-selectin in complement-dependent adhesion of human leukocytes to porcine endothelial cells. AB - BACKGROUND: Rapid leukocyte adherence to donor organ vasculature is a hallmark of hyperacute xenograft rejection. However, the molecular interactions required for leukocyte binding to vascular endothelium have not been characterized. METHODS AND RESULTS: Binding assays performed between human neutrophils and porcine aortic endothelial cells (PAEC) after exposure to human complement demonstrated that adhesion was mediated by both surface-bound C3b and C5b-9 activity. C5b-9 dependent adhesion was blocked by neuraminidase treatment of the neutrophils, suggesting that this binding was mediated by porcine P-selectin. Porcine P selectin was isolated from a PAEC cDNA library. The porcine P-selectin primary sequence contained an open reading frame encoding 646 amino acids with 82% identity to human P-selectin. Recombinant soluble porcine P-selectin specifically bound to human neutrophils and HL-60 cells. Transfection of COS cells with the full-length porcine P-selectin cDNA resulted in surface expression of the protein and markedly increased the binding of human neutrophils to these cells. The binding of both soluble and COS-expressed porcine P-selectin to human neutrophils was blocked by pretreatment of the neutrophils with neuraminidase or the addition of EDTA. Finally, treatment of PAEC with human thrombin or normal human serum but not purified human C5a- or C8-deficient human serum resulted in the rapid expression of porcine P-selectin on the cell surface. CONCLUSIONS: This report establishes that porcine P-selectin supports the binding of human neutrophils to PAEC in vitro. Further, these data suggest that sublytic deposition of C5b-9 during hyperacute rejection results in the expression of porcine P-selectin, which may contribute to the rapid adhesion of neutrophils to porcine xenografts. PMID- 10836379 TI - Requirement for early donor cell chimerism during prolonged survival of murine skin allografts. AB - BACKGROUND: Posttransplantation infusion of donor bone marrow cells (BMC) can prolong allograft survival in antilymphocyte antibody- (ALA) treated recipients. This study examined the hypothesis that chimerism of donor BMC origin contributes to allograft unresponsiveness. METHODS: Survival of day 0 skin grafts from C3H (H2k) donors was prolonged on ALA-treated B6AF1 mice by day +7 infusion of BMC from C3H or C3H-H2o2 (H2K(d)I(d)D(k)) mice. To test for functional chimerism, depletional anti-H2Kd antibody was injected at intervals after C3H-H2o2 BMC infusion. To confirm the persistence of active cells in the recipients, cells harvested from bone marrow of ALA- and C3H BMC-treated primary recipients were transferred to secondary ALA-treated recipients. Other recipients were infused on day +21 with additional donor BMC or light-density BMC that had been cultured in the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4) to promote the differentiation of dendritic cells (DC). RESULTS: Injection of depletional antibody targeting infused BMC interrupted skin graft survival, whether the injection was made on day +7, 2 hr after BMC injection, or as late as day +28. Prolonged graft survival was transferred to secondary recipients with cells recovered from primary recipient marrow as long as 4 weeks after initial donor BMC treatment. Graft survival prolonged by day +7 BMC was further enhanced by infusion on day +21 of light density cultured BMC, although a second dose of unfractionated BMC was inactive. Repeated injection of cultured BMC at 2-week intervals after day +21 prolonged graft survival even further. CONCLUSIONS: These data directly demonstrate that at least short-term donor BMC-derived microchimerism is required for prolonged allograft survival. The data further suggest that the active chimeric cells persist in recipient marrow. Finally, the beneficial effect of late infusion of cultured cells containing partially differentiated DC suggest that DC may be the active chimeric cells. PMID- 10836380 TI - High dose bone marrow transplantation induces deletion of antigen-specific T cells in a Fas-independent manner. AB - BACKGROUND: Monoclonal antibody induced tolerance to high doses of multiple lymphocyte stimulating (MLS)+minor mismatched bone marrow has recently been associated with clonal deletion, as reported in fully allogeneic models of bone marrow transplantation. FasL-induced apoptosis has been shown to mediate antigen specific T cell deletion after antigenic stimulation in wild-type and T cell receptor transgenic mice. Therefore, we investigate a role for the Fas pathway in deletional tolerance to high dose bone marrow. METHODS: Fas mutant and control mice (H-2k, MLS-1b) were tolerized under the cover of monoclonal antibodies to high dose (5 x 10(7) cells) AKR (H-2k, MLS-1a) bone marrow. Tolerance was confirmed by AKR skin grafting after antibody clearance. Antigen-reactive cell deletion was monitored by Vbeta6+ T cell elimination, measured by flow cytometry of peripheral blood throughout the experiment. Donor T cell (Thy1.1+) chimerism was assessed in a similar manner. RESULTS: Fas mutant mice infused with high dose AKR bone marrow under the cover of antibody were tolerant, as demonstrated by indefinite survival of AKR skin grafts. When high levels of donor cell chimerism were established in Fas mutant mice, peripheral deletion of antigen-reactive cells was observed to be independent of signaling through Fas. CONCLUSIONS: Apoptosis mediated by Fas receptor signaling is not the mechanism of clonal deletion of antigen-reactive cells after antibody facilitated high dose marrow transplantation. However, the Fas mutation does impair the development of adequate donor chimerism. PMID- 10836381 TI - Dominant tolerance and linked suppression induced by therapeutic antibodies do not depend on Fas-FasL interactions. AB - BACKGROUND: Nonlytic anti-CD4 monoclonal antibody therapy can be used to induce transplantation tolerance in rodent models. Such tolerance is often associated with dominant regulation, mediated by CD4+ cells, and characterized by infectious tolerance and linked suppression. Understanding the mechanisms by which CD4+ regulatory cells function may improve the manner in which current immunosuppressants are applied and may lead to the development of new tolerance inducing therapeutics. Fas-mediated apoptosis has been characterized as an important mechanism of peripheral self-tolerance and we here examine whether it has any role in anti-CD4 monoclonal antibody-induced dominant tolerance. METHODS: Tolerance to transplanted skin and bone marrow, mismatched for multiple minor histocompatibility antigens, was induced in Fas mutant and control mice using anti-CD4 and anti-CD8 monoclonal antibodies. To test for linked suppression, animals were transplanted with a second graft-bearing tolerated and third party antigens. The ability of splenocytes from tolerant animals to suppress graft rejection was assessed by transfer into partially immunocompromised recipients. RESULTS: Monoclonal antibody therapy rendered Fas mutant mice tolerant of minor disparate skin and bone marrow. Splenocytes from these and control tolerant animals when transferred into partially immunocompromised Fas mutant or control recipients, induced antigen-specific suppression of graft rejection. Additionally, tolerant Fas mutant mice accepted grafts bearing tolerated and third party antigens. CONCLUSIONS: Signal transduction through the Fas receptor plays no essential role in the induction of tolerance using anti-CD4 and anti-CD8 monoclonal antibodies or its maintenance by active regulation. PMID- 10836382 TI - Fas ligand gene transfer prolongs rat renal allograft survival and down-regulates anti-apoptotic Bag-1 in parallel with enhanced Th2-type cytokine expression. AB - BACKGROUND: Fas ligand (FasL) induces apoptosis of cells bearing Fas receptor, and may play a role in the acquisition of immune privilege. We have previously shown that adenovirus (Ad)-mediated FasL gene transfer significantly prolongs survival in a strongly major histocompatibility complex-incompatible rat kidney allograft model. This study analyzes putative mechanisms of FasL-mediated effects, with particular emphasis on Th1 and Th2 immune activation and Bag-1 expression, a Bcl-2-binding anti-apoptotic protein. METHODS: Kidney transplants were performed in Wistar-Furth to Lewis rat combination. Donor kidneys were perfused in situ with Ad-FasL or Ad-beta-Gal, and then transplanted. Kidney allografts were harvested at days 2, 7, and 56 and were evaluated by hematoxylin and eosin and immunohistochemical staining. The expression of FasL, Bag-1, and Th1/Th2 cytokine genes was assessed by Northern blots, Western blots, and competitive template reverse-transcriptase polymerase chain reaction, respectively. RESULTS: Intragraft expression of FasL was enhanced, whereas that of anti-apoptotic Bag-1 gene was diminished throughout, in Ad-FasL-transduced well-functioning renal allografts, compared with Ad-beta-Gal-treated rejecting controls. In parallel, the expression of mRNA coding for IL-2 and IFN-gamma remained depressed, whereas that of IL-4 and IL-10 reciprocally and progressively increased in the Ad-FasL animal group. CONCLUSIONS: Prolonged survival in Ad-FasL transduced rat renal allograft model correlates with down-regulation of Bag-1, a novel anti-apoptotic gene, and preferential Th2-type cytokine elaboration profile at the graft site. Because Th1-like cells are sensitive to FasL-mediated cytotoxic effects, T-cell apoptosis may preferentially spare Th2-like cells, with resultant prolonged graft survival. PMID- 10836383 TI - Immunomodulatory effects of pretransplant donor blood transfusion on T-cell independent xenoreactive immunity. AB - BACKGROUND: Pretransplant blood transfusions have beneficial effects on both clinical and experimental allograft survival. In the present study, we examined whether pretransplant hamster blood transfusions (pHBT) alone or together with peritransfusion immunosuppressive strategies designed to target B cells and/or natural killer (NK) cells, could modulate T cell-independent (T-I) xenoreactivity in athymic nude rats. METHODS: Hamster or mouse hearts were heterotopically xenotransplanted into untreated or treated athymic nude rats receiving either pHBT, anti-B cell or anti-NK cell therapy alone or their combinations. Xenoreactive antibodies (xAbs) and the percentage of NK cells were analyzed by FACScan analysis. NK cytotoxicity was measured by a standard 4 hr 51Cr release assay. Xenografts (Xgs) were examined by hematoxylin-eosin (H&E), by light microscopic method with Masson's trichrome and orcein staining, by immunofluorescent staining for immunoglobulin M and C3 deposition, and by immunohistochemical staining for infiltration of NK cells and macrophages (Mphis). RESULTS: In 1 of 6 rats given pHBT alone 2 weeks before receiving hamster xenografts, Xg survival was prolonged to 55 days compared with 3.0+/-1.2 days in the other 5 animals and with 3.0+/-0.6 days in untreated animals. In the 55 days, surviving Xg infiltration of Mphis and NK cells was seen together with severe signs of chronic rejection, such as fibrosis and obliterative vasculopathy. The addition of the anti-B cell immunosuppressant MNA715 (malononitriloamide x920715, 20 mg/kg/day) from day -14 to day +14 or of 100 microL of rabbit anti-asialo GM1 serum ([anti-ASGM1] an NK cell depleting antibody) on day -14 resulted in a significant and species-specific prolongation of the survival of hamster Xgs, respectively 59.8+/-9.6 days and 58.2+/-14.7 days (P<0.001 vs. control group), but not of mouse heart Xgs that were rejected in a normal tempo. All prolonged hamster Xgs were infiltrated with Mphis and NK cells and developed severe lesions of chronic rejection, such as fibrosis and obliterative vasculopathy. In contrast, MNA715 or anti-ASGM1 alone had no effect on Xg survival (4.8+/-1.7 days and 2.7+/-0.6 days, respectively). Combined MNA715/anti-ASGM1 treatment only moderately promoted Xg survival (10+/-5.0 days; P<0.001). A simultaneous administration of pHBT, MNA715, and anti-ASGM1 induced indefinite and species-specific Xg survival in all recipients. In vivo and in vitro studies demonstrated that both T-I B cell and NK cell species-specific xenotolerance were achieved. CONCLUSIONS: Pretransplant blood transfusion may have a species-specific immunomodulatory effect on T-I xenoreactivity. This effect is further enhanced by a temporary co-administration of MNA715 or by a single injection of anti-ASGM1. A combination of pHBT, MNA715, and anti-ASGM1 induces species-specific T-I xenotolerance. PMID- 10836384 TI - Cholesterol as an independent predictor of outcome after renal transplantation. AB - BACKGROUND: The debate on the role of high serum cholesterol levels in cardiovascular disease or chronic vascular rejection in kidney-transplanted patients has not yet been settled. METHODS: We studied the influence of serum cholesterol at 1 year after transplantation on the failure risk in all 676 kidney graft recipients who survived with a functioning graft. Other variables included in this analysis were donor/recipient age and gender, original disease, race, number of HLA-A and -B mismatches, previous transplants, postmortal or living related transplantation, and transplantation year. At 1 year after transplantation, we included: serum cholesterol, serum creatinine, proteinuria, and hypertension. RESULTS: In the Cox proportional hazards analysis, serum cholesterol at 1 year after transplantation turned out to be an important, independent variable influencing all end points (adjusted for all other variables in the model). The influence on graft failure censored for death was log-linear, and there was interaction with serum creatinine at 1 year. The adverse effect of elevated serum cholesterol levels on the graft failure rate decreased with increasing serum creatinine levels. The influence of serum cholesterol on the rate ratio (RR) for patient failure was linear too, and here there was interaction with recipient age. The negative influence of serum cholesterol on the RR for patient failure decreased with increasing recipient age. The risk for over-all graft failure was influenced by increasing serum cholesterol levels, and there was interaction with recipient age. Because recipient age had interaction with donor age and serum creatinine, the influence of all four variables together on the RR was estimated. It is shown that whereas the RR for over-all graft failure in young recipients of a renal transplant increases significantly with higher cholesterol levels, there is very little influence on the RR of elderly recipients. The risk increases proportionally with increasing serum creatinine levels. CONCLUSION: Serum cholesterol levels have an independent influence on graft, patient, and over-all graft failure. PMID- 10836385 TI - A comparison of fetal and adult porcine islets with regard to Gal alpha (1,3)Gal expression and the role of human immunoglobulins and complement in islet cell cytotoxicity. AB - BACKGROUND: It is still debated whether fetal or adult porcine islets should be the preferred choice for future clinical islet xenotransplantation. Each type of islet preparation has advantages and disadvantages compared with the other. Here we present a direct comparison between fetal and adult porcine islets with regard to Gal alpha(1,3)Gal expression, immunoglobulin and complement binding, and cytotoxicity after exposure to fresh human serum. METHOD: Islet single cell suspensions were prepared from adult and fetal islets by trypsin digestion. Fluorescein isothiocyanate-conjugated Bandeiraea simplicifolia isolectin B4 (BS IB4) and affinity-purified chicken anti-Gal alpha(1,3)Gal antibody was used to detect Gal alpha(1,3)Gal expression. Immunoglobulin and complement binding to the islet cells and cytotoxicity for islet cells was compared after incubation with fresh and heat-inactivated human sera and with an immune serum from a diabetic patient who received a fetal porcine islet transplant. Furthermore, two pools of human AB sera were depleted of porcine endothelial cell cytotoxic human anti-Gal alpha(1,3)Gal antibodies by absorption and were used to analyze the effect of Gal alpha(1,3)Gal antibody removal on islet cell cytotoxicity. RESULTS: Fetal islet cells readily bound both BS-IB4 and the chicken anti-Gal alpha(1,3)Gal antibody. None of 10 adult porcine islet preparations were stained by BS-IB4. In comparison, IgY anti-Gal Ab binding was detected in two of eight adult islet isolations, whereas the other six preparations showed marginal/no binding. After incubation of fetal islet cells with fresh human serum, C3c binding was strongly positive and IgM binding variable, with occasional binding of IgG and no detectable binding of IgA. Adult islet cells were also strongly positive for C3c but did not bind detectable amounts of IgM, IgG, or IgA. Immune sera from a patient who had received fetal porcine islets showed the presence of induced antibodies that bound to fetal islet cells and to porcine peripheral blood lymphocytes, whereas binding to adult islet cells was barely detectable. Fresh human sera showed a high and similar level of complement-mediated lytic activity for both adult islet cells (78+/-22%) and fetal islet cells (75+/-16%). Cytotoxicity for fetal islet cells and peripheral blood lymphocytes was significantly reduced when the corresponding sera were depleted of anti-Gal antibodies before use (P=0.002 and P=0.003, respectively). In contrast, no difference in cytotoxicity for adult islet cells was detected when anti-Gal depleted human sera were used. CONCLUSION: Gal alpha(1,3)Gal expression is occasionally detectable on adult porcine islet cells, but not as readily and at a lower level, compared with fetal islet cells. Thus, as porcine fetal islets mature to adult islets, the expression of the Gal alpha(1,3)Gal epitope gradually diminishes. Consequently, cytotoxic anti-Gal alpha(1,3)Gal antibodies in human serum play an important role in the lysis of fetal but not adult porcine islet cells. PMID- 10836386 TI - Quantitative polymerase chain reaction assessment of chimerism in non-human primates after sex-mismatched islet and bone marrow transplantation. AB - BACKGROUND: Accurate assessment of chimerism in recipients of islet and bone marrow transplantation (BMT) may allow for a clearer assessment of the role of chimerism in islet engraftment or rejection. A quantitative polymerase chain reaction (PCR) assay was developed for the detection of the sex-determining region of the Y chromosome (SRY) in peripheral blood samples from female non human primate recipients of allogeneic male islets and vertebral body marrow (VBM) from the same donor. METHODS: The assay incorporates a synthetic internal standard (IS) containing the same primer template sequences as the target to compete for primer annealing and amplification. Each DNA sample was coamplified with a constant amount of IS. The concentration of male DNA in the test samples was calculated from the regression equation of a standard curve that was generated by plotting the logarithm of the ratio of the intensities of SRY to IS PCR products versus the logarithm of known percentages of input male DNA. RESULTS: This method allows for a correction of the variability of efficiency of the PCR technique and also overcomes the drawback of time-consuming competitive PCR. Using this assay, we quantitated the amount of male DNA in samples taken from female baboon recipients of male islets and VBM. There was detectable male donor DNA in the samples taken one day after BMT; pre-BMT samples were negative. This technique works well for samples obtained from rhesus and cynomogus monkeys as well. CONCLUSIONS: It is a practical method for accurately evaluation of chimerism after sex-mismatched allogeneic BMT in non-human primate models. PMID- 10836387 TI - Human herpesvirus-6 and human herpesvirus-7 in the bone marrow from healthy subjects. AB - BACKGROUND: Human herpesviruses (HHVs) 6 and 7 are recently discovered betaherpesviruses. Although HHV-6 has been associated with disordered hematopoiesis in bone marrow transplant recipients, little information is available on the presence of both viruses in the bone marrow from healthy subjects. METHODS: We detected HHV-6 and HHV-7 DNA by means of polymerase chain reaction in bone marrow and peripheral blood samples from 18 healthy subjects who underwent total hip arthroplasty. RESULTS: Genomic HHV-6 and HHV-7 DNA were detected in 11% and 67% of the blood samples, respectively, and in 28% and 50% of the bone marrow samples, respectively. CONCLUSIONS: Both viruses may be present in the bone marrow without hematopoiesis disorder and can be transmitted through bone marrow infusion. Therefore, the causative role of these two viruses in some bone marrow diseases cannot be inferred simply from the detection of their genome in bone marrow by means of polymerase chain reaction. PMID- 10836388 TI - Comparison between tacrolimus and cyclosporine as immunosuppressive agents compatible with tolerance induction by CD4/CD8 blockade. AB - BACKGROUND: Donor-specific tolerance can be induced in mice by transient antibody blockade of the CD4 and CD8 co-receptors of T cells. To evaluate the potential application of CD4/CD8 blockade in the clinic, we have asked if either tacrolimus or cyclosporine counteract the tolerogenic process. METHODS: Using the fully mismatched mouse cervical heart transplant model, BALB/c (H2d) to CBA (H2k), the experimental groups were (i) no therapy, (ii) tacrolimus (1 mg/kg, i.p., daily, days 0-14); (iii) cyclosporine (25 mg/kg, i.p., daily, days 0-14), (iv) blocking monoclonal antibodies (mAbs) to CD4 and CD8 (2 mg, i.p., starting day 0 and on alternating days thereafter for a total of six doses), (v) tacrolimus plus mAbs, and (vi) cyclosporine plus mAbs. RESULTS: Allograft survival was prolonged in both the tacrolimus and cyclosporine groups. mAbs alone induced tolerance, and mAbs combined with tacrolimus also induced tolerance. In contrast, the combination of mAbs and cyclosporine was toxic. CONCLUSIONS: The induction of tolerance by blocking CD4 and CD8 was not prevented by tacrolimus. However, combination of cyclosporine with the same tolerogenic protocol was toxic to mice. PMID- 10836389 TI - Withdrawal of mycophenolate mofetil in stable renal transplant recipients. AB - BACKGROUND: Mycophenolate mofetil (MMF) has been demonstrated to decrease episodes of acute rejection in renal transplant recipients during the first year after transplantation. The utility of MMF after 1 year is less clear. METHODS: Forty-five stable renal transplant recipients on maintenance therapy of cyclosporine microemulsion, MMF, and prednisone had MMF withdrawn at approximately 1 year after transplantation. A matching concurrent group of 45 stable renal transplant recipients served as the case control group. RESULTS: Two of 45 patients in the MMF withdrawal group suffered an acute rejection episode as opposed to 1 of 45 in the control group. Both patients who rejected in the withdrawal group had adequate cyclosporine levels and had no recent decrease in prednisone dose. There was no evidence of an increased incidence of proteinuria or increased creatinine levels in the MMF withdrawal group. CONCLUSION: In general, withdrawal of MMF in stable renal transplant recipients is well tolerated. No increased risk of rejection could be demonstrated in this patient group. A larger study will be needed to confirm our result. PMID- 10836390 TI - Evolution of attitudes at U.S. transplant centers toward kidney donation by friends and altruistic strangers. AB - A severe shortage of organs is one of the major barriers facing transplantation today. One of the proposals designed to overcome this serious problem is to increase the use of genetically unrelated living kidney donors. Excellent results have been achieved with these volunteers and cogent arguments have been made that this practice is ethically acceptable. These considerations have encouraged many transplant centers to break with tradition and accept spousal donors. To see if there has been a similar change in attitudes toward other types of unrelated living donors, a survey was mailed to 208 U.S. renal transplant centers; 129 (62%) were returned. Ninety-three percent of responding centers said they would accept a close friend as a kidney donor. Although the majority of centers would not consider an altruistic stranger, a sizeable minority (38%) would. When compared with the results of previous surveys, these data show that attitudes toward unrelated living kidney donors have gradually become much more liberal. PMID- 10836391 TI - Absence of the left portal vein: a difficulty for reduction of liver grafts? AB - BACKGROUND: Absence of the portal bifurcation is exceptional and characterized by an absent extrahepatic portal vein bifurcation, the right portal vein only being at the porta hepatis. There is no extraparenchymal left portal vein. This may represent a problem in liver splitting, reduction, and living related transplantation. METHOD: A case was encountered during reduction of a cadaveric liver allograft to a left lateral segmental graft from a 40-kg cadaveric donor to a 15-kg recipient. The portal venous inflow was reconstructed with a vein graft via a novel extrahilar approach to the left portal vein at the umbilical fissure. RESULTS: This graft was used successfully in a 3-year-old child requiring transplantation for a failed Kasai operation for extrahepatic biliary atresia. The child is now well, 1 year posttransplant, after an uneventful postoperative course with good portal flow within the graft. CONCLUSION: The situation of an absent left portal vein extrahepatic course should not preclude splitting or reduction procedures. The innovative technical solution, we propose, should add to the armamentarium of the liver transplant surgeon contemplating a left lateral segmental graft for the paediatric liver transplant recipient. PMID- 10836392 TI - Real-time monitoring of cytomegalovirus infections after stem cell transplantation using the TaqMan polymerase chain reaction assays. AB - BACKGROUND: Real-time monitoring of cytomegalovirus (CMV) infections in transplant patients demands a rapid and high-throughput CMV DNA quantification method. METHODS: TaqMan polymerase chain reaction assays based on CMV immediate early protein exon 4 and glycoprotein B were developed and were compared with a COBAS AMPLICOR CMV MONITOR (CMM) test for quantifying CMV DNA in peripheral blood leukocytes from seven stem cell transplant patients having received antiviral treatment. RESULTS: There was a good correlation between the TaqMan assays and the CMM test for CMV DNA quantification. The throughput of the TaqMan assays was, however, about 3 times higher than that of the CMM test. The CMV DNA dynamics patterns determined by the TaqMan polymerase chain reaction were well in line with the outcome of the antiviral therapy. CONCLUSIONS: The TaqMan assays may potentially serve as a useful tool for rapid quantification of CMV infections in stem cell transplant patients. PMID- 10836393 TI - Conversion from tacrolimus to cyclosporine in stable renal transplant patients: safety, metabolic changes, and pharmacokinetic comparison. AB - BACKGROUND: Although conversion between tacrolimus and cyclosporine has been performed when indicated for rejection or adverse effects, the safety and metabolic outcome of elective conversion from tacrolimus to cyclosporine has not previously been examined. METHODS: Conversion from tacrolimus to cyclosporine was performed in 19 recipients of cadaver renal transplants at 3-6 months after transplantation. Pharmacokinetic profiles and biochemical studies were performed three times, in steady state, before, and after conversion. RESULTS: Patient and graft survival was 100% at 3 months after conversion, with no rejection episodes. Three patients have been subsequently converted back to tacrolimus, two for rejection and one for hirsutism. There were no significant changes in creatinine, urate, or blood sugar levels after conversion, but the mean plasma magnesium rose from 0.73 (0.63-0.97) to 0.82 (0.65-1) mmol/L (P=0.037), and the mean plasma cholesterol rose from 5.2 (3.4-6.8) to 5.5 (3.8-7.6) mmol/L (P=0.033). Pharmacokinetic profiles were measured before and after conversion, and showed that cyclosporine (Neoral) exhibited significantly less interpatient and intrapatient variability than tacrolimus, for area under the curve (AUC), maximum concentration after dose (Cmax), minimum concentration after dose (Cmin), and time to maximum concentration (Tmax). CONCLUSION: This is the first study that has examined the outcome of conversion from tacrolimus- to cyclosporine-based immunosuppression in stable patients after renal transplantation. This conversion was performed without early immunological hazard, but there was a small rise in blood cholesterol levels after conversion. Pharmacokinetic studies showed that cyclosporine in the form of Neoral exhibited less inter- and intrapatient variability than tacrolimus, although this is of uncertain clinical significance. PMID- 10836394 TI - Additional interferon alpha for lamivudine resistant hepatitis B infection after liver transplantation: a preliminary report. AB - Resistance formation is a major problem in antiviral treatment of hepatitis B recurrence after liver transplantation. One possible therapeutic approach is an antiviral combination therapy with synergistic drugs. Four patients who were transplanted for chronic hepatitis B were analyzed retrospectively. All patients had reinfection of the graft and breakthrough of hepatitis B virus (HBV) during consecutive famciclovir and lamivudine monotherapy. Subsequently a combination therapy of lamivudine and interferon alpha 2a (3 times 3 million units weekly) was initiated. Addition of interferon markedly reduced viral replication rate in all patients. Three patients became HBV-DNA negative despite lamivudine resistance, but only two had a sustained response. No patient seroconverted to anti-HBe or lost HBsAg, but all patients showed a normalization of alanine aminotransferase and aspartate aminotransferase levels. No severe complications, and especially no rejection episodes occurred. Therefore lamivudine combined with interferon might be used for the therapy of hepatitis B reinfection after liver transplantation. PMID- 10836395 TI - Successful transplantation of highly selected CD34+ peripheral blood stem cells in a HLA-sensitized patient treated with immunoadsorption onto protein A. AB - BACKGROUND: Haploidentical bone marrow transplantation with preexisting anti-HLA antibodies is associated with a high risk of graft failure. METHODS: A 27-year old female patient with chronic myeloid leukemia and evidence of several osseous chloromas had no suitable matched bone marrow donor, and fluorescence cytometric cross-match (FCXM) revealed antibodies against donor-specific HLA-molecules. Immunoadsorption onto staphylococcal protein A was applied to remove these antibodies, and peripheral stem cell transplantation was performed from her haploidentical sister after a negative FCXM was documented after immunoadsorption and conditioning treatment. RESULTS: FCXM for donor lymphocytes and stem cells remained negative throughout the posttransplant period, and engraftment of donor cells was documented on day +69. CONCLUSION: Immunoadsorption onto protein A should be considered in stem cell transplantation even from an haploidentical donor where anti-HLA antibodies and a positive FCXM are documented. PMID- 10836396 TI - Local secretion of a chimeric anti-CD4 antibody protects against graft rejection in the NOD mouse. AB - BACKGROUND: Engineering a graft to secrete its own immunosuppressive antibodies may minimize the risks associated with current high dose systemic immunosuppression. METHODS AND RESULTS: A beta cell insulinoma cell line (NIT-1) was transfected with genes encoding a chimeric anti-CD4 antibody. The NIT-1 cells secreted functional chimeric anti-CD4 antibody that bound to the CD4 molecule on mouse thymocytes and inhibited in vitro proliferation of CD4+ve T cells. Both test and control transfected cell lines grew at a similar rate in immunodeficient mice. In immunocompetent NOD mice, NIT-1 cells are normally rejected by a cellular immune response against the SV40 T antigen. Although control transfected NIT-1 cells were rapidly rejected by NOD mice, anti-CD4 secreting NIT-1 cells grew significantly better and were able to form tumors at the site of injection. CONCLUSIONS: The local secretion of chimeric anti-CD4 antibody from transfected cells can contribute to graft survival in our transplantation model. PMID- 10836397 TI - Chronic renal allograft dysfunction: a role for mycophenolate mofetil? PMID- 10836398 TI - Influence of blood sampling sites on measurement of blood cyclosporine concentration. PMID- 10836399 TI - Neutrophils respiratory burst is not impaired under cyclosporine treatment following liver transplantation. PMID- 10836400 TI - Mycophenolate mofetil in the prevention of rejection after kidney transplantation. PMID- 10836401 TI - Is PERV transfer across hollow fiber membranes relevant to bioartificial liver model? PMID- 10836402 TI - From dating to biophysics--20 years of progress in applied ESR spectroscopy. AB - ESR spectroscopy represents a tool for quantitative radiation analysis that was developed somehow simultaneously for dating purposes in Japan and in Germany for high-level standardization, in the mid-seventies. Meanwhile, ESR dosimetry has reached an established metrology level. Present research fields of ESR dosimetry consider post-accident dose reconstruction in the environment, and biophysical dosimetry using human tissues. The latter promises a re-definition of radiation risk for chronicle exposure to be derived from individuals of the early nuclear facilities in Russia, and hopefully United States in the future. An attempt is made to sketch development and potential future of the ESR technique. PMID- 10836404 TI - Accuracy considerations in EPR dosimetry AB - A brief overview of some of the potential sources of inaccuracies that are common to all kinds of electron paramagnetic resonance (EPR) dosimetry is given. The instrumental sources discussed are inaccuracies of receiver gain calibration, instability of the spectrometer sensitivity, and neglect of inhomogeneity of the fields in the resonators. The discussed sources of inaccuracies related to data processing are violations of basic assumptions of the used version of the least squares technique, incorrect model selection for fitting the data, and nonoptimal design of the calibration experiment. Methods for detecting and correcting such inaccuracies are briefly discussed and references to the original literature are given. PMID- 10836403 TI - In vivo EPR dosimetry of accidental exposures to radiation: experimental results indicating the feasibility of practical use in human subjects. AB - Low frequency electron paramagnetic resonance (EPR) provides the potential advantage of making accurate and sensitive measurements of absorbed radiation dose in teeth in situ, i.e. without removing the teeth from the potential victim. The potential limiting factors for making such measurements are: (1) whether low frequency EPR is sufficiently sensitive to detect radiation-induced signal in human teeth; (2) whether sufficient sensitivity can be maintained under in vivo conditions. In this manuscript, we summarize results indicating that this approach is feasible. Using 1.2 GHz EPR spectroscopy, we found that the lower limit for these measurements in isolated human teeth is 0.2 Gy or lower. Measurements of radiation-induced EPR signals in the teeth of living rats were achieved with sufficient sensitivity to indicate that, when taking into consideration the larger mass of human teeth, similar measurements in human teeth in situ would provide sensitivity in the dose range for potential accidental exposures. We estimate that the current lower limit for detecting radiation doses in human teeth in situ (in vivo) is 0.5-1.0 Gy; this would be sufficient for determining if a person has been exposed to potentially life threatening doses of ionizing radiation. The limiting factor for sensitivity appears to be background signals rather than signal/noise, and there are feasible means to overcome this problem and further increase sensitivity. The additional instrumental developments required to make an effective in vivo EPR dosimetric spectrometer for the measurements in teeth in human subjects in situ, seem quite achievable. PMID- 10836405 TI - Differentials or integrals: pluses and minuses in their application to additive dose techniques AB - Retrospective radiation dosimetry is based on the growth of radiation sensitive defects. In Electron Paramagnetic Resonance (EPR), the peak-to-peak measurement of the differential of the absorption spectrum is used, whereas in luminescence dosimetry, the response of the sample to irradiation is traditionally determined by the height or the integral of the emission spectrum. It has been shown experimentally and can be proved mathematically that, except in special cases, the environmental dose estimate (De) depends on which parameter is used to measure radiation response. Since integral methods are directly related to the number of radiation-produced defects, it is increasingly assumed that they will give the best estimates of De in EPR as well as in luminescence dosimetry. We show that the disparity in De is not due to any inherent superiority of a particular method, but is rather due to the different degree of interference between wide and narrow peaks in the different methods. In spite of their conceptual nicety, integral methods do not necessarily give the best estimate of De. This is demonstrated by computer simulation and guidelines are also given on selecting the best method in any particular case. Differential techniques might be very profitably applied to luminescence dosimetry as well as EPR dosimetry. The differential spectrum not only shows detail which may be obscured in the integral curve, but also provides a very simple graphical means of isolating a preferred spectral component. This leads to the possibility of extending the range of substances which may be reliably used for dosimetry or dating by luminescence techniques. PMID- 10836406 TI - Comparison of sample preparation and signal evaluation methods for EPR analysis of tooth enamel. AB - In dose reconstruction by EPR dosimetry with teeth various methods are applied to prepare tooth enamel samples and to evaluate the dosimetric signal. A comparison of seven frequently used methods in EPR dosimetry with tooth enamel was performed. The participating Institutes have applied their own procedure to prepare tooth enamel samples and to evaluate the dosimetric signal. The precision of the EPR measurement and the dependence of the estimated dosimetric signal with irradiation up to 1000 mGy were compared. The obtained results are consistent among the different methods. The reproducibility of the dosimetric signal and its estimated relation with the absorbed dose was found to be very close for the applied methods with one possible exception. PMID- 10836407 TI - Preliminary report on the development of a virtually nondestructive additive dose technique for EPR dosimetry. AB - We propose a new approach to the additive dose method in EPR dosimetry studies for tooth enamel specimens. We outline a specialized routine whereby the sample may be left for the most part unirradiated, while only a small aliquot of the sample will be additively irradiated to relatively large doses. The routine is done in such a way so as not to significantly compromise either precision or accuracy of the dose reconstruction. It is also demonstrated that the overall throughput of the dose reconstruction is not appreciably compromised. With this potential ability, the utility of an international dose/sensitivity standard for EPR dosimetry of teeth is considered. PMID- 10836408 TI - Temperature stability of carbonate groups in tooth enamel. AB - The tooth enamel EPR signal at g = 1.9973 consists of several components, which correspond to different functions of the spatial orientation of the CO2- radicals, have different temperature stability and different saturation curves. Using a new technique described in this paper, we have detected and investigated two groups of CO2- radicals with different thermal stability and saturation characteristics. PMID- 10836409 TI - Anisotropy effects of EPR signals and mechanisms of mass transfer in tooth enamel and bones. AB - Peculiarities of the internal construction of tooth enamel and bones that cause anisotropy effects and mass transfer in these objects are described. It is shown that the composition of the mineral component of teeth and bones depends on a mechanical-electrical mechanism, which pumps ions into nanocrystals. Decrease in the efficiency of the mechanical-electrical mechanism results in demineralization of enamel and bones, which progresses most rapidly at a disease of the biomineral or under special conditions, such as in space flights. Effects of signal anisotropy in the practice of retrospective EPR dosimetry are discussed. PMID- 10836410 TI - Electron spin resonance (ESR) studies of CO2- radicals in irradiated A and B-type carbonate-containing apatites AB - ESR spectra of irradiated carbonate-containing hydroxyapatites are complex and strongly dependent on the method of preparation of these materials. This paper describes results of our studies of the ESR spectra of irradiated A- and B-type carbonated hydroxyapatites. The most intense and stable lines in these spectra are attributed to CO2- radicals. The ESR spectrum of A-type carbonated apatite can be predominantly due to either axial or orthorhombic CO2- species, depending on whether the apatite was cooled in or without a CO2 flow. The B-type carbonated apatite has an ESR spectrum composed basically of signals from isotropic and orthorhombic CO2- species. These signals are different in intensity, line width, and g-factor for samples heated for 3 h at 200, 400 and 600 degrees C. The effect of the method of preparation of the A- and B-type carbonated apatites on the ESR spectra and the application of these materials to the ESR dating and dosimetry are discussed. PMID- 10836411 TI - Gamma dose response of synthetic A-type carbonated apatite in comparison with the response of tooth enamel. AB - Synthetic A-type carbonated apatite samples were irradiated at room temperature with 60Co gamma rays. Their ESR spectra consist of the lines of CO2- and CO3- radicals of orthorhombic and axial symmetry. The measurements carried out immediately after sample irradiation showed that CO2- species are produced by decomposition of CO3- radicals. Intensity of the CO2- lines in the synthetic and enamel samples increases during the first 400 and 200 h after irradiation, respectively. The dependence of the EPR signal on the dose varies with carbonate content of the sample. The dose response curve for tooth enamel is steeper for the synthetic material. PMID- 10836412 TI - Generation and conversion of electronic defects in calcium carbonates by UV/Vis light AB - Radical species like CO2-, CO3-, SO2-, and SO3- can be created by exposing natural and synthetic calcium carbonates to sunlight or to the light of a Hg(Xe) lamp. This poses as a risk for ESR dating of these materials, because the radicals formed by light exposure cannot be distinguished from those generated by radioactivity. Furthermore, paramagnetic centers like SO2- electrons trapped near Zn2+ or Cd2+ ions, surface defects, and radicals with g' = 2.0040, can be bleached in gamma-irradiated samples by light and show conversion effects. PMID- 10836413 TI - The role of cytogenetics in early triage of radiation casualties. AB - Preliminary dose estimates by chromosomal analysis can be made rapidly in order to supplement early triage of radiation casualties based on clinical signs. An in vitro simulation of an accident with many casualties receiving whole or partial body exposure in the range 0-8 Gy is described. Faced with an urgent need for rapid results, confirmation of clinical triage can generally be obtained from scoring 20 metaphases per subject. Scoring should be increased to 50 cells where there is disagreement with the initial assessments or evidence of significantly inhomogeneous exposure. PMID- 10836414 TI - The IAEA's co-ordinated research project on biodosimetry, 1998-2000. PMID- 10836415 TI - Perspective on biological dosimetry from the aspect of individual radiosensitivity: the context of DNA double-strand breaks and chromosomal aberrations. PMID- 10836416 TI - Tailed nuclei and dicentric chromosomes in irradiated subjects. AB - A study of peripheral blood smears from irradiated Chernobyl liquidators and other subjects has shown nuclei of some lymphocytes to have a protrusion into the cytoplasm. Such abnormal nuclei are called "tailed" nuclei (TN). Sixteen main morphologic types, observed in human peripheral blood lymphocytes, are described. The frequency of appearance of lymphocytes with TN in a group of Chernobyl liquidators was significantly higher than in control groups (p < 0.001, in all cases). A positive correlation was found between the TN frequency in lymphocytes and dicentric chromosomes in lymphocytes cultured to metaphase (p < 0.001). Elevated frequencies of dicentrics, higher than 0.1%, were found in 17 out of 22 subjects in whom the frequency of lymphocytes with TN was 0.8% and more. Abnormalities of the TN type in lymphocytes are likely to result from breakdown of chromosome bridges formed by dicentrics. The TN can be considered as a possible marker of irradiation and, therefore, their detailed study is important. PMID- 10836417 TI - Reciprocal translocations as an indicator for radiation exposures in the low dose range. AB - A few years back, the frequency of dicentrics was determined shortly after exposure in five accidentally exposed radiation workers. In all cases the observed dicentric yield was significantly higher in comparison with the background level, and the resulting estimated whole body doses lay between 0.2 and 0.3 Gy. Now, a number of years later (1 to 11 years), the frequency of translocations has been determined by means of the FISH technique. Chromosomes 2, 4 and 8 were painted. The measured translocation frequency lay, however, within the range of the spontaneous variation between individuals. No radiation exposure could, therefore, be proven. In two further cases, the dicentrics were determined by means of the conventional Giemsa staining technique, and the translocations by means of chromosome painting carried out on the same blood samples, which were taken about 3 and 10 years, respectively, after the radiation exposure. The dose estimates obtained on the basis of dicentric frequency using the Qdr method, and on the basis of FISH detected translocations, are compared. PMID- 10836418 TI - Correlation of the LET-dependent TL-response of LIF:Mg,Ti TL-dosemeters and gentoxic endpoints after proton irradiation. AB - The high temperature ratio (HTR) method using LiF:Mg,Ti thermoluminescent dosemeters allows in addition to the measurement of absorbed dose, the evaluation of the biological effectiveness of radiation. In order to analyse the correlation of the measured parameter HTR and gentoxic endpoints irradiation of cell cultures and TLDs were performed in a 62 MeV proton beam. The gentoxic effects respectively the relative biological effectiveness RBE, show an excellent correlation with the HTR due to a similar dose and LET-dependent behaviour. PMID- 10836419 TI - Retrospective dose estimation using the dicentric distribution in human peripheral lymphocytes. AB - The paper presents the results of retrospective dose estimations for a number of Chernobyl victims by different approaches to the analysis of dicentric distributions in peripheral lymphocytes in blood samples taken a long time after the accident. Retrospective dose estimations are compared with dose estimates in the same individuals made immediately after the accident by dicentric frequencies. PMID- 10836420 TI - Determination of somatic mutant frequencies at glycophorin A and T-cell receptor loci for biodosimetry of acute and prolonged irradiation. AB - Somatic mutant frequencies at glycophorin A (GPA) and T-cell receptor (TCR) loci were assessed. The dependence of the GPA mutant frequency on doses of acute and prolonged irradiation was shown. In the case of acute irradiation the GPA mutant frequency displayed a three-fold greater dose-related increase as compared to prolonged irradiation. A dose-dependent increase in the TCR variant frequency was found only in a group of subjects with recent exposures. In Chernobyl clean-up workers the TCR mutant frequency was significantly higher than in control non irradiated individuals. PMID- 10836421 TI - Novel data set for retrospective biodosimetry using both conventional and FISH chromosome analysis after high accidental overexposure. AB - Blood samples of ten Chernobyl and one non-Chernobyl victims were analysed both by conventional cytogenetics and by fluorescence in situ hybridization (FISH) using a cocktail of chromosomes 2, 3 and 8. The analysed group comprised men acutely irradiated mainly in 1986 and aged 26-47 years at the time of first blood sampling. All of them displayed acute radiation syndrome of varying severity. Chromosome analysis of the earliest blood samples was carried out by conventional scoring of unstable aberrations with the number of metaphases analysed per individual ranging from 35 to 300. Estimated individual doses ranged from 0.85 to 9.8 G y. After a 10 year delay, i.e. in 1996, blood samples were analysed both by conventional scoring of unstable aberrations and by FISH measurements of stable ones. Usually about 500 metaphases per individual were scored. Estimated by the FISH-method individual translocation (tc + ti) frequencies ranged from 2.2 to 116.8 per 100 cells full genome equivalent. Based on three different published dicentric dose response, in vitro curves individual doses were calculated from the earliest dicentric frequencies. A dose response curve for truly persisting translocations (tc + ti) was estimated over the range 1-10 Gy. PMID- 10836422 TI - Stable and unstable aberrations in lymphocytes of Chernobyl accident clearance workers carrying rogue cells. AB - Cells with multiple chromosomal aberrations, the so-called rogue cells, were found in blood samples from more than 100 Chernobyl accident clearance workers. A comparative analysis of frequencies of stable and unstable chromosomal aberrations in two worker groups--those with or without rogue cells was made. A higher level of unstable aberrations in persons carrying rogue cells was observed. No difference in the level of stable aberrations between the groups was seen. The possibility of low dose alpha irradiation causing the chromosomal damage is raised. PMID- 10836423 TI - Cytogenetic investigation of occupationally irradiated persons a long time after exposure. AB - Chromosomal analyses were performed in blood lymphocytes of 33 persons previously occupationally exposed to ionizing radiation. An increased frequency of chromosome aberrations, mainly dicentrics, a long time, up to 47 years after exposure, suggests that it is possible to use the method of unstable aberration analysis for assessment of previous radiation exposure. The enhanced yield of dicentrics was observed in persons who may have received irradiations above permitted limits. The aberrations represent a residue from a higher initially induced incidence. They can not be used reliably for retrospective dosimetry but do provide a marker for old exposure. PMID- 10836424 TI - Dosimetry studies in Zaborie village. AB - Dosimetry studies in Zaborie, a territory in Russia highly contaminated by the Chernobyl accident, were carried out in July, 1997. Studies on dosimetry for people are important not only for epidemiology but also for recovery of local social activity. The local contamination of the soil was measured to be 1.5-6.3 MBq/m2 of Cs-137 with 0.7-4 microSv/h of dose rate. A case study for a villager presently 40 years old indicates estimations of 72 and 269 mSv as the expected internal and external doses during 50 years starting in 1997 based on data of a whole-body measurement of Cs-137 and environmental dose rates. Mean values of accumulated external and internal doses for the period from the year 1986 till 1996 are also estimated to be 130 mSv and 16 mSv for Zaborie. The estimation of the 1986-1996 accumulated dose on the basis of large scale ESR teeth enamel dosimetry provides for this village, the value of 180 mSv. For a short term visitor from Japan to this area, external and internal dose are estimated to be 0.13 mSv/9d (during visit in 1997) and 0.024 mSv/50y (during 50 years starting from 1997), respectively. PMID- 10836425 TI - Cytogenetic features of leukaemias diagnosed in residents of areas contaminated after the Chernobyl nuclear accident. AB - A comparison of chromosomal abnormalities in bone marrow leukaemic cells and of stable and unstable aberrations in lymphocytes of patients with hematological malignancies who live in areas with or without contamination by the Chernobyl nuclear accident has been made using FISH and G-banding. Healthy residents of these areas comprised the control group. No systematic cytogenetic differences of leukaemic cells between patients from contaminated and uncontaminated areas were observed. Lymphocyte aberrations, however, were generally higher in all subjects from contaminated areas. Comparison has been made with specific cytogenetic features of leukaemic cells and a high level of stable aberrations in lymphocytes of patients with secondary leukaemias that had developed after chemo- and/or radio-therapy. PMID- 10836426 TI - IAEA high-dose intercomparison in 60Co field. International Atomic Energy Agency AB - Nine high-dose calibration laboratories collaborated with the IAEA in a 'double blind' intercomparison of absorbed dose in the 10-50 kGy dose range. Bureau International des Poids et Mesure undertook the analysis of the results as well as assisted in the development of the protocol. IAEA alanine-ESR transfer dosimeters were used for this intercomparison. The agreement was within 2.0% at 15 kGy and 2.2% at 45 kGy which is very good considering the variety of irradiation conditions employed by the participants. The mean of the dose values measured by the IAEA Dosimetry Laboratory was within 1% of the mean of the dose values stated by the participants for both dose values. PMID- 10836427 TI - An automated system for the measurement of alanine/EPR dosimeters AB - NPL for several years has offered mailed reference dosimetry services based on alanine/EPR dosimeters, both at industrial and therapy dose levels. Compared to other methods of reference dosimetry, operator involvement in alanine/EPR has been found to be relatively high, and contributes significantly to the overall economics of the process. Commercially available sample changers are not suitable for high accuracy applications, and it has proved necessary to develop a dedicated automation system to handle NPL alanine dosimeter pellets. In this paper we describe an automatic sample changer for placing and retrieving alanine pellets into and out of the cavity of a standard research grade EPR spectrometer. Up to 32 pellets can be held in each removable sample tray. The sample changer software has been interfaced into the spectrometer control software to enable complete automation of the measurement process, including the optimization of spectrometer settings and rotation of the sample within the cavity. PMID- 10836428 TI - The mechanisms of radical formation in L-alpha-alanine AB - Modeling of radical transformations in L-alpha-alanine after irradiation was performed for isolated radicals and for clusters. Special attention was devoted to the explanation of the experimental results concerning selective proton transfer and behavior of cation-radicals because a unique interpretation of the corresponding experiments is very difficult. Both semi-empirical and ab initio methods were used depending on the size of system under investigation. The results obtained show the usefulness of the computer simulation for processes in rather complex materials used in dosimetry. PMID- 10836429 TI - ESR/Alanine gamma-dosimetry in the 10-30 Gy range. AB - We report Alanine Dosimeter preparation, procedures for using the ESR/Dosimetry method, and the resulting calibration curve for gamma-irradiation in the range from 10-30 Gy. We use calibration curve to measure the irradiation dose in gamma irradiation of human blood, as required in Blood Transfusion Therapy. The ESR/Alanine results are compared against those obtained using the thermoluminescent dosimetry (TLD) method. PMID- 10836430 TI - Transferability of ASTM/NIST alanine-polyethylene recipe at ISS. American Society for Testing and Materials/National Institute for Standards and Technology. Istituto Superiore de Sanita AB - Alanine-polyethylene solid state dosimeters were prepared at Istituto Superiore di Sanita (ISS) following the recipe proposed by National Institute of Standards and Technology (NIST) with the goal of testing its transferability. Dosimeters were prepared using 95% alanine and 5% polyethylene, by weight. They are rugged and of increased sensitivity, repeatability and reproducibility as respect to the ISS alanine-paraffin pellets. Reproducibility of about 1% was obtained at 10 Gy and at 3 Gy if one single pellet or a stack of five dosimeters were used, respectively. PMID- 10836431 TI - Electron arc therapy treatment planning verification with alanine/EPR dosimetry. AB - Alanine/EPR dosimetry was applied to measure dose distributions during electron arc therapy treatments generated by electron beams moving isocentrically over a stated arc. Alanine-polyethylene pellets, prepared at ISS according to the NIST recipe, inserted in a homogeneous cylindrical and anthropomorphic phantoms, were used. Preliminary, alanine response to static electron beams in the (6-20) MeV nominal energy range was studied. Then alanine dosimetry was applied to determine the dose versus the gantry angle at a reference point in a cylindrical homogeneous phantom. Finally, arc therapy treatment planning verification was performed in anthropomorphic phantom. PMID- 10836432 TI - Strategy for finding new materials for ESR dosimeters AB - The right strategy for finding a new ESR dosimetric material sensitive to radiation is to follow the orthodox procedures used in the development of thermoluminescence dosimeters (TLD) and phosphorescence studies. Modern procedures used in materials sciences, such as computer calculation of molecular orbitals (MO), should be employed to estimate the ESR and optical properties of prospective materials. Radiation effects in lithium and magnesium sulfates and metal salts of organic acids, such as lithium and magnesium lactates, have been investigated in search for tissue-equivalent dosimeter with a large G value. PMID- 10836433 TI - Inorganic EPR dosimeter for medical radiology. PMID- 10836434 TI - A decomposition study of the EPR spectrum of irradiated sucrose AB - In general, the EPR spectra of irradiated sugars are very complex because of their multicomponent character. In this study we applied a multivariate statistical method called MLCFA, maximum likelihood common factor analysis, and it predicted at least six components contributing to the total EPR spectrum of irradiated sucrose. Three dominant components have already been isolated in an irradiated sucrose single crystal using electron nuclear double resonance (ENDOR) and ENDOR induced EPR (EI-EPR). Results of EPR simulations based on the ENDOR data are in a reliable agreement with the experimental EPR spectra of irradiated sucrose single crystals. PMID- 10836435 TI - ESR and optical absorption studies of gamma- and electron-irradiated sugar crystals AB - Electron spin resonance (ESR) studies of the free radicals induced in gamma- or electron-irradiated sugar crystals were performed. The number of radicals increases linearly, pointing out that this material can be used as a dosimeter. The optical absorption studies show the presence of several distinctive bands in the infrared and UV region, whose intensity changes with the irradiation dose. An interpretation of these results in terms of the formation of free radicals and possible crosslinking along the sugar molecular chains is presented. PMID- 10836436 TI - Development of ammonium tartrate as an ESR dosimeter material for clinical purposes. AB - The crystalline substance ammonium tartrate is investigated with respect to its suitability as a clinical dosimetry material. The properties investigated are: the radical stability for absorbed doses relevant for clinical use, the improvement in sensitivity when the crystals are deuterated and the linear electron transfer (LET)-dependence. After photon irradiation to an absorbed dose of 20 Gy, the signal increases rapidly during the first 6 h. After this period, the changes are more moderate and can be corrected for. The signal-to-noise ratio of irradiated ammonium tartrate is twice the corresponding value of alanine. By deuterating the crystals, the sensitivity can be further improved by a factor of 1.4. As expected, the signal decreases with increasing LET of the radiation, but no changes in the spectrum shape was observed. PMID- 10836437 TI - A new ESR dosimeter based on polytetrafluoroethylene AB - A new electron spin resonance (ESR) dosimetry system based on the ESR response of polytetrafluoroethylene is presented. The dose response of these dosimeters irradiated with 60Co gamma radiation was linear in the range from 6 x 10(2) up to 1.5 x 10(5) Gy, having a repeatability of +/-1% and a high degree of reliability (Cf = 1.054). The persistence of the signal was monitored over a period of one month. ESR signal decay was 5-7% in the temperature range from room temperature up to 40 degrees C. PMID- 10836438 TI - Metal ion-organic compound for high sensitive ESR dosimetry AB - A systematic study to find a tissue equivalent and high sensitive dosimeter material has been made to stimulate the field of ESR dosimetry. Lithium acetate dihydrate (Li-Ac.2H2O:CH3COOLi.2H2O) and lithium phosphate (Li-phosphate:Li3PO4) were irradiated by gamma-rays to study radicals with (ESR) in addition to magnesium lactate (Mg-lactate (CH3CH(OH)COO)2Mg) doped with nominal pure lithium lactate (Mg(Li)-lactate) and lithium lactate (CH3CH(OH)COOLi) doped with Mg lactate (Li(Mg)-lactate). A triplet spectrum with intensity ratio of 1:2:1 in Li Ac.2H2O was ascribed to acetate radical which has g = 2.0031+/-0.0004 and hyperfine splitting of A/gbeta = 2.12+/-0.1 mT. The Li-phosphate spectrum shows splitting due to anisotropic g-factors of g(parallel) = 2.0190+/-0.0005 and g(perpendicular) = 1.9974+/-0.0004. Quartet spectra with the intensity ratio of 1:3:3:1 in Mg(Li)-lactate and Li(Mg)-lactate were ascribed to lactate radicals with g-factors of 2.0032+/-0.0004 and 2.0029+/-0.0004 and the intensity ratio of 1:3:3:1 and A/gbeta = 1.92+/-0.06 and 1.82+/-0.06 mT, respectively. The response to gamma-ray dose and the thermal stability as well as the effect of UV illumination have been studied. The obtained number of free radicals per 100 eV (G-values) were 0.4+/-0.13, 1.02+/-0.31, 1.35+/-0.35 and 0.78+/-0 for Li-Ac.2H2O, Li-phosphate, Mg(Li)-lactate, and Li(Mg)-lactate, respectively. The lifetimes were estimated from Arrhenius plots to be approximately 2.0+/-0.6, 50.7+/-20 and 10+/-3.5 years for Li-phosphate, Mg(Li)-lactate and Li(Mg)-lactate, respectively. The lifetime for Li-Ac.2H2O cannot be estimated because of the decomposition by heating. PMID- 10836439 TI - ESR of environmental objects from Semipalatinsk Nuclear Test Site PMID- 10836440 TI - Radiation effects in dry ice: models for a peak on the Arrhenius curve AB - Dry ice (solid CO2) occurs in the polar caps of Mars, on the surface of Triton, and in places in the outer planets of our solar system. Radicals in gamma irradiated solid CO2 have been studied by ESR for future applications of ESR dating on outer planets. The annealing curves for CO3- radical (ESR signal at g = 2.0126) can be described neither by the first-order nor the second-order decay kinetics. The peak observed in the Arrhenius plot can result from two parallel first-order kinetic processes. Radicals that provide overlapping signals are CO3- (g1 = 2.0057, g2 = 2.0126, g3 = 2.0161; activation energy E = 0.10 eV; frequency factor v0 = 4 x 10(1) s(-1)) and HO2 (g1 = 2.0040, g2 = 2.0055, g3 = 2.0360), which have E = 0.28 eV and v0 = 7 x 10(5) s(-1)). Hence, HO2 is more thermally stable, and use of HO2 is promising for ESR dating. PMID- 10836441 TI - Current issues on EPR dose reconstruction in tooth enamel. AB - This paper focuses on the opportunities to gain risk assessment information from populations exposed to ionizing radiation. Details are presented on the scale of the nuclear events in several areas within the former Soviet Union. An overview of the issues associated with dose reconstruction by electron paramagnetic resonance (EPR) is given. Principles and outline of the EPR tooth enamel dose assessment are described. Technical details, the latest achievements and the problems of each procedural step are analyzed. In addition, the present accuracy of the EPR dose reconstruction, its problems and cost are discussed. PMID- 10836442 TI - Application of EPR retrospective dosimetry for large-scale accidental situation. AB - Above 3000 tooth enamel samples, collected at population of radioactive contaminated territories after Chernobyl accident, the Chernobyl liquidators, the retired military of high radiation risk and the population of control radiation free territories were investigated by EPR spectroscopy method in order to obtain accumulated individual exposure doses. Results of EPR spectra measurements are stored in data bank; enamel samples are also stored in order to provide the possibility to repeat the measurements in future. Statistical analysis of results has allowed to detect the contribution into EPR signal in tooth enamel due to the action of the natural background radiation, and the radioactive contamination of territory. In general, the average doses of external exposure of the population obtained with EPR spectroscopy of teeth enamel are consistent with results based on other methods of direct and retrospective dosimetry. Essential exceeding of the individual doses above the average level within the population groups was observed for some persons. That gave the possibility to detect the individuals with overexposure, which were included into groups for medical monitoring. PMID- 10836443 TI - Some aspects of EPR dosimetry of liquidators. AB - Doses of few hundred liquidators were reconstructed using EPR dosimetric technique, developed and routinely used in SCRM. Both cumulative and accidental dose values were determined. Obtained results are analyzed and discussed. PMID- 10836444 TI - New computer procedure for routine EPR-dosimetry on tooth enamel: description and verification. AB - A software code was developed for fitting the EPR tooth enamel spectrum using linear combinations of Gaussian functions. The software is fastening EPR dose reconstruction for routine applications. The verification of the software was done comparing with the selective saturation method as an independent procedure of signal evaluation. Both methods were applied for the dose reconstruction of 13 teeth mostly from the Techa riverside. The evaluated doses by the two methods demonstrate an excellent agreement. PMID- 10836445 TI - Tooth enamel EPR dosimetry: sources of errors and their correction. AB - Some of the most important sources of systematic errors in dose determination using tooth enamel EPR spectroscopy and ways of reducing those errors are discussed. Enamel from the outside of the front teeth should not be used for dose determination because of induction of paramagnetic centers by solar light. The accuracy of the method in the low dose range is limited by variation in the shape of the EPR signal of unirradiated enamel, which can be described by an initial intrinsic signal and which varies for different samples with standard deviation of 20-30 mGy. The energy dependence of enamel sensitivity should be taken into account in the form of a correction factor. The value of this factor is estimated at 1.1-1.3 for real radiation fields in radiation contaminated territories. Variation in enamel sensitivity for different samples is shown to be within limits of 10-15% of the average value. PMID- 10836446 TI - Electron spin resonance dosimetry of teeth of Goiania radiation accident victims. AB - Electron spin resonance (ESR) spectroscopy is used to assess absorbed doses of six teeth belonging to victims of the highly irradiated group of Goiania accident. The influence of the broad background signal at g = 2.0040 as well as of the unstable fraction of CO2- radicals was taken into account in dose estimates. Three victims teeth showed absorbed doses comparable to those estimated by chromosomal analysis. For the other three teeth, the doses were higher by a factor of 1.3, 1.8 and 2.2. PMID- 10836447 TI - Metabolism in tooth enamel and reliability of retrospective dosimetry. AB - Most of the radiation centers in tooth enamel and bones are formed by the reactions COOH + gamma-->CO2- + H+, and CO2 + e- -->CO2-. These centers can be easily changed by metabolism and have a low stability in living organisms. The efficiency of CO2- radical production by irradiation is substantially different for teeth and bones in living rat, dead rat, and teeth and bone removed from rat. It is shown that the suitability of tooth enamel for retrospective dosimetry can be evaluated from the anisotropy of its EPR signals. PMID- 10836448 TI - ESR dating: is it still an 'experimental' technique? AB - Nearly 25 years ago, Motoji Ikeya demonstrated the potential of ESR dating. From a single substance (stalagmitic carbonate) and a single site (Akiyoshi Cavern), the field has grown to include materials from all over the world and time periods from a few thousand years ago to several million years ago. A vigorous program of instrumentation development has increased the precision of measurements as well as opening up new ways of collecting and interpreting spectra. Yet there are still references to ESR dating as an 'experimental' technique, one which cannot be trusted to produce dates that are accurate or precise. This paper discusses areas for which this is true and suggests what should be done to convince skeptics. Other areas for which the evidence suggests that ESR is at least as reliable as 'standard' methods will also be covered. PMID- 10836449 TI - EPR spectrum deconvolution and dose assessment of fossil tooth enamel using maximum likelihood common factor analysis. AB - In order to determine the components which give rise to the EPR spectrum around g = 2 we have applied Maximum Likelihood Common Factor Analysis (MLCFA) on the EPR spectra of enamel sample 1126 which has previously been analysed by continuous wave and pulsed EPR as well as EPR microscopy. MLCFA yielded agreeing results on three sets of X-band spectra and the following components were identified: an orthorhombic component attributed to CO2-, an axial component (CO3(3-)), as well as four isotropic components, three of which could be attributed to SO2-, a tumbling CO2- and a central line of a dimethyl radical. The X-band results were confirmed by analysis of Q-band spectra where three additional isotropic lines were found, however, these three components could not be attributed to known radicals. The orthorhombic component was used to establish dose response curves for the assessment of the past radiation dose, D(E). The results appear to be more reliable than those based on conventional peak-to-peak EPR intensity measurements or simple Gaussian deconvolution methods. PMID- 10836450 TI - ESR study of elephant tooth enamel from the Karlich-Seeufer site in Germany. AB - Enamel from 6 different positions in a well preserved elephant tooth from the Karlich-Seeufer site in Germany has been irradiated up to 32 kGy. The X-band (v = 9.5 GHz) ESR spectra of two subsamples have been decomposed into three real components with Maximum Likelihood Common Factor Analysis (MLCFA). One of these components due to orthorhombic CO2- radicals is predominant. Dose response curves for the contributions of these MLCFA components and for different heights in the ESR spectra have been obtained and fitted with different models. Depending on the model, the equivalent dose for the preferably used height at g = 1.9973, due to CO2-, ranges from 70 to 130 Gy. Due to a very low uranium and thorium content in both enamel and dentine (< or = approximately 10 ppb) and to an important external y-attenuation, the ages fluctuate between 300 and 575 ka. PMID- 10836451 TI - Improvements in dating tooth enamel by ESR. AB - ESR dating requires that growth curves be determined by interpreting complex spectra. Spectra, however, can vary significantly in shape and field position between different samples, or occasionally between subsamples, even though the mineralogy remains the same. In some cases, this spectral variability does not affect the resulting accumulated dose calculation. In other cases, signal subtraction may be needed. However, some samples that until recently might have been considered unsuitable for dating are now shown to yield accurate and precise results because a broad interference peak is integral to the hydroxyapatite signal. By studying the spectrum at the Q-band frequency, it can be shown that the interfering signal in most cases is not a problem for dating. A second concern has been that artificially irradiating sample aliquots can introduce a short-lived component that is simply an unstable enhancement of the dating signal. The apparent accumulated dose from growth curves created immediately after irradiation is considerably greater than that after annealing, although the curve's shape remains unchanged. Annealing both the natural and artificially irradiated signal shows the dating signal's lifetime to be greater than 10(10) years. PMID- 10836452 TI - ESR dating of a toxodon tooth from a Brazilian karstic cave. AB - A pre-molar of a Toxodon platensis (sp), a large mammal that inhabited the forests of Southeast Brazil during quaternary times, was dated by electron spin resonance (ESR). Enamel and dentine were separated giving, the following archeological doses (AD): (4.0+/-0.8) and (11+/-2) Gy, respectively. This difference was explained by the higher content of Uranium in dentine (25.1+/-0.3) ppm as compared to enamel (0.37+/-0.03) ppm. Taking account of the internal dose contribution to the AD, the age obtained is (6.7+/-1.3) ky BP for dentine and (5.0+/-1.6) ky BP for enamel. PMID- 10836453 TI - Formation and decay of the E1' center and of its precursor AB - The E1' center has been used for ESR dating of quartz with assuming that the signal intensity increases with natural radiation dose as those of other ESR signals do. However, this simple assumption is not necessarily correct. Formation and decay of the E1' center are closely related with its precursor, diamagnetic oxygen vacancies. Gamma ray of large dose (> 100 kGy) creates oxygen vacancies giving little dose rate effect, which, therefore, might be useful for dating of granites and high dose dosimetry. PMID- 10836454 TI - Bleaching of ESR signals by the sunlight: a laboratory experiment for establishing the ESR dating of sediments AB - A laboratory bleaching experiment was performed in order to improve the method of ESR dating of sediments. Several sedimentary, volcanic, and granitic quartz samples showed consistent bleaching response for the exposure to halogen lamps. It was found that the most sensitive signals are the Ti-H and Ti-Na centers. There was no difference observed within the samples exposed to light filtered by several color glass plates, according to the present preliminary result. PMID- 10836455 TI - ESR dating of a quartz single crystal from the Menderes Massif in Turkey AB - Electron spin resonance (ESR) has been employed to date the most recent geological metamorphism by use of a large quartz single crystal extracted from the metamorphic region of Menderes Massif (MM) in Western Turkey. The ESR spectra showed two paramagnetic centers: Al and Ti centers. The former was used for dating, since the latter was already saturated by natural radiation. The age, estimated to be 1.8+/-0.5 Ma, does not agree with the previous age evaluated by isotopic dating techniques suggesting that the thermal or geothermal stressing history of the metamorphic region must have changed the apparent ESR age. A detailed annealing experiment at various temperatures was performed to investigate the influence of thermal annealing in the past and the resulting life time was calculated to be 8.4 x 10(4) a for the Al center, the lifetime for Ti being shorter than that for Al. The lifetime of Al is thus shorter than its ESR measured age. The discrepancy can be explained in terms of the present-day annual dose or 238U, 232Th, and 40K, concentrations being much smaller than the previous annual doses under severe environmental conditions, such as erosive, natural chemical etching and weathering processes. PMID- 10836456 TI - Temperature stabilization of alanine dosimeters used for food processing and sterilization AB - The International Atomic Energy Agency has established a dose quality audit service for radiation processing facilities. The objective of the service is to provide an independent check on the routine dosimetry system in use at the facility. The audit service is based on the use of alanine EPR dosimetry. Generally, alanine dosimeters are irradiated at the facility together with a product, and the response is then analyzed at the IAEA laboratory. Practice of the audit service has shown that the main uncertainty in alanine dosimetry is due to absence of temperature control at the irradiation facilities. Here, a method for stabilizing the temperature of the dosimeter during irradiation is proposed. PMID- 10836457 TI - Identification and dose determination using ESR measurements in the flesh of irradiated vegetable products AB - The international commerce of vegetable products is often dependent on the quarantine protections that are imposed by the importing countries because of the fear of contamination by fruit flies. The use of ionizing radiation as a treatment for these products can be used to remove this problem and a real proof of irradiation can contribute to the implementation of the international commerce. ESR measurement on the pulp of vegetable products can be used as a proof of irradiation using the species introduced in cellulose that are found uniquely in irradiated products. The stability of these species are compatible with the life of the products analyzed. The pulp signal intensity is sufficient to identify products irradiated with doses as low as 100 Gy for some fruits. PMID- 10836458 TI - Electron paramagnetic resonance identification of irradiated cuttlefish (Sepia officinalis L.). AB - Gamma-ray irradiated fresh cuttlefish bone display very intense Electron Paramagnetic Resonance (EPR) spectra, which could be attributed to five different centers, including CO2-, previously observed in irradiated carbonates. All centers display a positive correlation with the absorbed dose, described by linear or saturation type dependencies. During 36 h of isothermal annealing at 100 degrees C, three of them decayed with time exponentially, one remained constant, while the EPR spectrum amplitude of the fifth increased. PMID- 10836471 TI - Objectivity in the assessment of equine welfare. PMID- 10836473 TI - Effect of age and training on murmurs of atrioventricular valvular regurgitation in young thoroughbreds. AB - Cardiac auscultation was carried out on 111 Thoroughbred horses age 2-5 years to test the hypothesis that athletic training might influence the development of atrioventricular (AV) valve regurgitation in young Thoroughbreds. Murmurs of valvular regurgitation were identified and graded on a 1-6 scale. There were 2 sources of auscultation data: 1) 55 2-year-old horses that were examined by auscultation before training commenced and 9 months later when at race fitness; 2) 56 horses age 2-5 years that were examined on one occasion only (25 2-year olds, 23 3-year-olds, five 4-year-olds and five 5-year olds). All horses in the second data set were in full training and racing regularly at the time of the examination. To conclude the study, 35 horses were selected randomly from both groups of horses and examined with colour-flow Doppler echocardiography. The aim of the final part of the study was to check specificity and sensitivity of auscultation for detection of AV valve murmurs and therefore validate the auscultation findings. Prior to training, the prevalence in 2-year-old racehorses of murmurs of mitral regurgitation and tricuspid regurgitation was 7.3% (4/55) and 12.7% (7/55), respectively. After training, the prevalence proportions increased to 21.8% (12/55) and 25.5% (14/55). After training, one horse developed a murmur characteristic of aortic regurgitation. The differences in murmur prevalence were statistically significant for mitral and tricuspid regurgitation (paired t test results: mitral regurgitation, P = 0.019; tricuspid regurgitation, P = 0.007), as were the differences in mean murmur grade (P = 0.018 and P = 0.0006, respectively). There were no significant effects of age on the prevalence of valvular regurgitation in 56 horses examined at race fitness. Auscultation was a specific (specificity 100%) and reasonably sensitive method for detection of murmurs of mitral and tricuspid regurgitation (mitral regurgitation: positive predictive value 100%, negative predictive value 84%, tricuspid regurgitation: positive predictive value 100%, negative predictive value 65%). These data suggest that the prevalence and grade of murmurs of mitral and tricuspid valvular regurgitation increase in 2-year-old Thoroughbreds after 9 months of athletic training. Whereas the effects of age and growth on the prevalence of murmurs cannot be ruled out from these data, this study suggests that there is an influence of athletic training on the development of atrioventricular valvular regurgitation in flat-racing Thoroughbreds. PMID- 10836472 TI - Equine dental disease part 4: a long-term study of 400 cases: apical infections of cheek teeth. AB - Of 400 horses referred because of equine dental disease, 162 suffered from primary apical infections of their cheek teeth (CT), including 92 with maxillary CT infections and 70 with mandibular CT infections. Maxillary swellings and sinus tracts were more common (82 and 26% incidence, respectively) with infections of the rostral 3 maxillary CT, than with infections of the caudal 3 maxillary CT (39 and 5% incidence, respectively). Nasal discharge was more commonly present with caudal (95%) than rostral (23%) maxillary CT infections. Mandibular CT apical infections commonly had mandibular swellings (91%) and mandibular sinus tracts (59%) and these infections were closely related to eruption of the affected CT. A variety of treatments, including medical treatment, apical curettage, repulsion and oral extraction of affected teeth were utilised in these cases, with oral extraction appearing to be most satisfactory. Infections of caudal maxillary CT with a secondary paranasal sinusitis were most refractory to treatment, with a complete response to the initial treatment achieved in just 33% of these cases. Most other cases responded fully to their initial treatment. The long-term response to treatment was good in most cases. PMID- 10836474 TI - Respiratory responses of mature horses to intravenous lobeline bolus. AB - The respiratory stimulant lobeline has been used in equine clinical practice to increase inspiratory and expiratory airflow rates at rest in order to facilitate investigation of both lower and upper airway function. Some of the responses to lobeline in the pony have been reported, but the detailed time course, effect of dose, possible side effects and reproducibility associated with lobeline administration have not been described in the horse. Respiratory airflow rates and oesophageal pressure were measured with a Fleisch No. 5 pneumotachometer and lightweight facemask and a microtip pressure transducer catheter, respectively. The output of the Fleisch pneumotachometer was calibrated for flow rates up to +/ 70 l/s. Seven mature horses with no clinical signs of respiratory disease were studied. Investigations were conducted to determine: (1) the responses to different doses of lobeline (0.15, 0.20, 0.25 and 0.30 mg/kg bwt) as a rapid i.v. bolus (6 horses); (2) arterial blood gases during and after lobeline administration (0.20 mg/kg bwt; 3 horses); and (3) the reproducibility of lobeline-stimulated hyperpnoea (5 horses; 2 doses of 0.20 mg/kg bwt lobeline, 15 min apart). All horses tolerated the lobeline-stimulated hyperpnoea well, although one always coughed or snorted at the onset. Mild tremor was noted following the highest dose in several horses. Apnoea of approximately 40 s was common after the hyperpnoea. Both tidal volume (VT) and frequency (fR) increased with lobeline dose. During peak hyperpnoea at a dose of 0.30 mg/kg bwt, peak inspired flow rate (PIF), peak expired flow rate (PEF) and minute ventilation (VE) were mean +/- s.e. 41+/-5 l/s, 61+/-10 l/s and 920+/-99 l/min, respectively. The hyperpnoea also caused marked changes in arterial PaO2, PaCO2 and pHa at 90 s after lobeline (0.20 mg/kg bwt) administration (mean +/- s.e. 146.0+/-6.9 mmHg, 20.6+/-0.8 mmHg and 7.707+/-0.020, respectively) compared to at rest (mean +/- s.e. 104.0+/-4.0 mmHg, 50.6+/-2.8 mmHg and 7.432+/-0.012). Dynamic lung compliance (Cdyn) was unaltered by lobeline administration. The lobeline-induced hyperpnoea was highly reproducible, with no significant difference in any of the parameters during 2 stimulations 15 min apart. Lobeline induced highly reproducible responses without any apparent adverse effects and may be useful in the investigation of pulmonary function in healthy horses and those with airway disease. PMID- 10836475 TI - Headshaking in horses: possible aetiopathogenesis suggested by the results of diagnostic tests and several treatment regimes used in 20 cases. AB - Twenty mature horses with typical headshaking of 2 week-7 year duration were studied. Clinical examinations included radiography of the head and nasopharyngeal endoscopy. All were assessed at rest and at exercise, both before and after fitting an occlusive nasal mask, application of tinted contact lenses and the perineural anaesthesia of the infraorbital and posterior ethmoidal branches of the trigeminal nerve. Infraorbital anaesthesia had no effect in 6/7 cases but 11/17 (65%) cases showed a 90-100% improvement following posterior ethmoidal nerve anaesthesia. Tinted contact lenses had no apparent long-term benefit, although 2 cases showed a transient improvement. We found no other evidence to suggest a photic aetiology in the current series of cases. Treatment regimens based on the results of the diagnostic investigative methods included sclerosis of the posterior ethmoidal branch of the trigeminal nerve. This was effective in some cases but the benefits were temporary. Cyproheptadine alone was ineffective but the addition of carbamazepine resulted in 80-100% improvement in 80% of cases. Carbemazepine alone was effective in 88% of cases but results were unpredictable at predefined dose rates. The positive response to carbamazepine, combined with the clinical features is consistent with involvement of the trigeminal nerve, particularly the more proximal branches such as the posterior ethmoidal nerve. Headshaking has some clinical features in common with trigeminal neuralgia in humans. As a result of the findings detailed in this paper, we conclude that a trigeminal neuritis or neuralgia may be the basis of the underlying aetiopathology of equine headshaking. Initial observations of the positive response of headshakers to carbamazepine therapy is encouraging. However, future studies will include a more detailed investigation of dosages, duration of effectiveness (in some cases it appears short-lived) and other effects. In practice there is a realistic possibility of controlling but not curing headshaking with carbamazepine therapy at the present time. Other future investigations will include details of the functional anatomy of the trigeminal nerve and the role of the P2 myelin protein in headshaking and other neurological disease. PMID- 10836476 TI - Functional adaptation of equine articular cartilage: the formation of regional biochemical characteristics up to age one year. AB - Biochemical heterogeneity of cartilage within a joint is well known in mature individuals. It has recently been reported that heterogeneity for proteoglycan content and chondrocyte metabolism in sheep develops postnatally under the influence of loading. No data exist on the collagen network in general or on the specific situation in the horse. The objective of this study was to investigate the alterations in equine articular cartilage biochemistry that occur from birth up to age one year, testing the hypothesis that the molecular composition of equine cartilage matrix is uniform at birth and biochemical heterogeneity is formed postnatally. Water content, DNA content, glycosaminoglycan content (GAG) and biochemical characteristics of the collagen network (collagen content, hydroxylysine content and hydroxylysylpyridinoline [HP] crosslinks) were measured in immature articular cartilage of neonatal (n = 16), 5-month-old foals (n = 16) and yearlings (n = 16) at 2 predefined differently loaded sites within the metacarpophalangeal joint. Statistical differences between sites were analysed by ANOVA (P<0.01), and age correlation was tested by Pearson's product moment correlation analysis (P<0.01). In neonatal cartilage no significant site differences were found for any of the measured biochemical parameters. This revealed that the horse has a biochemically uniform joint (i.e. the cartilage) at birth. In the 5-month-old foals and yearlings, significant site differences, comparable to those in the mature horse, were found for DNA, GAG, collagen content and hydroxylysine content. This indicates that functional adaptation of articular cartilage to weight bearing for these biochemical parameters takes place during the first months postpartum. Water content and HP crosslinks showed no difference between the 2 sites from neonatal horses, 5-month-old animals and yearlings. At both sites water, DNA and GAG decreased during maturation while collagen content, hydroxylysine content and HP crosslinks increased. We propose that a foal is born with a uniform biochemical composition of cartilage in which the functional adaptation to weight bearing takes place early in life. This adaptation results in biochemical and therefore biomechanical heterogeneity and is thought to be essential to resist the different loading conditions to which articular cartilage is subjected during later life. As collagen turnover is extremely low at mature age, an undisturbed functional adaptation of the collagen network of articular cartilage at a young age may be of significant importance for future strength and resistance to injury. PMID- 10836477 TI - Six intestinal microflora-associated characteristics in sport horses. AB - The aim of this study was to investigate 6 microflora-associated characteristics (MACs) in faecal samples from horses and to compare the results with baselines previously established in other mammals. A MAC is defined as any anatomical structure, physiological, biochemical or immunological characteristic in a host, which has been acted on by microorganisms. When the active microbes are absent, as in germ-free animals and healthy newborn organisms, the corresponding characteristic is defined as germ-free animal characteristic (GAC). The MACs studied were degradation of mucin, conversion of cholesterol to coprostanol and bilirubin to urobilinogens, inactivation of tryptic activity, degradation of beta aspartylglycine and excretion of short-chain fatty acids. Five MACs were studied in 19 sport horses and the mucin pattern investigated in 25 other sport horses. Our results show that conversion of cholesterol to coprostanol was present in all horses, irrespective of diet. All horses also demonstrated excretion of urobilinogens; however, the values varied depending on diet. Mucin and beta aspartylglycine were totally degraded, as occurs similarly in other animal species. Excretion of short-chain fatty acids was lower than values previously found in man, rats and pigs, and the level of faecal tryptic activity was low. Our results show that the MAC/GAC concept is as good in monogastric herbivores as in omnivores for investigations of endogenous and exogenous factors influencing the intestinal ecosystem(s). Therefore, use of the MAC/GAC concept has a potential significance for future studies on functions related to the microflora. PMID- 10836478 TI - Cyclical endometrial steroid hormone receptor expression and proliferation intensity in the mare. AB - The aims of this study were to investigate the steroid hormone receptor expression and the proliferation intensity during the equine endometrial cycle by immunohistological methods, established for routine examination of formalin fixed, paraplast-embedded specimens. Endometrial biopsy specimens were obtained during one cycle from 7 mares. In comparison with the blood steroid hormone levels the quantity and distribution of oestrogen (ER) and progesterone receptors (PR) and the proliferation marker Ki-67 antigen expression were investigated. Rising 17beta-oestradiol concentrations in preoestrus induce a synchronous expression of ER, PR and Ki-67 antigen in stromal cells. In the early dioestrus 17beta-oestradiol levels decrease and progesterone levels reach their maxima. This correlates with an intense proliferation activity and the highest hormone receptor expression in epithelial cells. In accordance to the morphological features of asynchronous glandular differentiation in fibrotic areas (endometrosis) their epithelial hormone receptor expression is out of phase. PMID- 10836479 TI - Study of intragastric administration of doxycycline: pharmacokinetics including body fluid, endometrial and minimum inhibitory concentrations. AB - The objectives of this study were to determine the pharmacokinetics and tissue concentrations of doxycycline after repeated intragastric administration, and to determine the minimum inhibitory concentrations (MIC) for equine pathogenic bacteria. In experiment 1, 2 mares received a single intragastric dose of doxycycline hyclate (3 mg/kg bwt). Mean peak serum concentration was 0.22 microg/ml 1 h postadministration. In experiment 2, 5 doses of doxycycline hyclate (10 mg/kg bwt), dissolved in water, were administered to each of 6 mares via nasogastric tube at 12 h intervals. The mean +/- s.e. peak serum doxycycline concentration was 0.32+/-0.16 microg/ml 1 h after the first dose and 0.42+/-0.05 microg/ml 2 h after the fifth dose. The mean trough serum concentrations were > 0.16 microg/ml. Highest mean synovial concentration was 0.46+/-0.13 microg/ml and highest mean peritoneal concentration was 0.43+/-0.07 microg/ml, both 2 h after the fifth dose. Highest urine concentration was mean +/- s.e. 145+/-25.4 microg/ml 2 h after the last dose. Highest endometrial concentration was mean +/- s.e. 1.30+/-0.36 microg/ml 3 h after the fifth dose. Doxycycline was not detected in any of the CSF samples. Mean +/- s.e. Vd(area) was 25.3+/-5.0 l/kg and mean t1/2 was 8.7+/-1.6 h. In experiment 3, minimum inhibitory concentrations of doxycycline were determined for 168 equine bacterial culture specimens. The MIC90 was < or = 1.0 microg/ml for Streptococcus zooepidemicus and 0.25 microg/ml for Staphylococcus aureus. Based on drug concentrations achieved in the serum, synovial and peritoneal fluids and endometrial tissues and MIC values determined in the present study, doxycycline at a dose of 10 mg/kg bwt per os every 12 h may be appropriate for the treatment of infections caused by susceptible (MIC < 0.25 microg/ml) gram-positive organisms in horses. PMID- 10836480 TI - Repeatability of Doppler ultrasound measurements of hindlimb blood flow in halothane anaesthetised horses. AB - The purpose of this study was to determine the repeatability of femoral blood flow recorded using Doppler ultrasound in anaesthetised horses. Doppler ultrasound of the femoral artery and vein was performed in 6 horses anaesthetised with halothane and positioned in left lateral recumbency. Velocity spectra, recorded using low pulse repetition frequency, were used to calculate time averaged mean velocity (TAV), velocity of component a (TaVa), velocity of component b (TaVb), volumetric flow, early diastolic deceleration slope (EDDS) and pulsatility index (PI). Within-patient variability was determined for sequential Doppler measurements recorded during a single standardised anaesthetic episode. Within-patient variability was also determined for Doppler and cardiovascular measurements recorded during 4 separate standardised anaesthetic episodes performed at intervals of at least one month. Within-patient variation during a single anaesthetic episode was small. Coefficients of variation (cv) were <12.5% for arterial measurements and <17% for venous measurements. Intraclass correlation coefficient was >0.75 for all measurements. No significant change was observed in measurements of cardiovascular function suggesting that within-patient variation observed during a single anaesthetic episode was due to measurement error. In contrast, within-patient variation during 4 separate anaesthetic episodes was marked (cv>17%) for most Doppler measurements obtained from arteries and veins. Variation in measurements of cardiovascular function were marked (cv>20%), suggesting that there is marked biological variation in central and peripheral observed. Further studies are warranted to determine the ability of this technique to detect differences in blood flow during administration of different anaesthetic agents. PMID- 10836481 TI - Cross-sectional study of faecal shedding of Giardia duodenalis and Cryptosporidium parvum among packstock in the Sierra Nevada Range. AB - Faecal specimens from 305 horses and mules used as packstock at one of 17 commercial or governmental (National Park Service, US Forest Service) operations were examined for Giardia duodenalis and Cryptosporidium parvum using immunofluorescent microscopy. Fourteen packstock (4.6%) were shedding G. duodenalis cysts, with herd-level prevalences ranging 0-22%. Number of packstock in the corral, size of corral and density of packstock in the corral were associated with the odds of shedding G. duodenalis cysts. None of the horses had detectable C. parvum oocysts. Assuming a sensitivity of at least 43% and a specificity of 100% for our assay, the estimated maximum true prevalence of shedding of C. parvum for packstock would be < or = 2.3% of the population. These data suggest that faecal dispersal of C. parvum on back country watersheds is unlikely with packstock. PMID- 10836482 TI - Frequency domain analysis of heart rate variability in horses at rest and during exercise. AB - The pattern of variation in heart rate on a beat-to-beat basis contains information concerning sympathetic (SNS) and parasympathetic (PNS) contributions to autonomic nervous system (ANS) modulation of heart rate (HR). In the present study, heart period (RR interval) time series data were collected at rest and during 3 different treadmill exercise protocols from 6 Thoroughbred horses. Frequency and spectral power were determined in 3 frequency bands: very low (VLF) 0-< or = 0.01, low (LO) >0.01-< or = 0.07 and high (HI) >0.07-< or = 0.5 cycles/beat. Indicators of sympathetic (SNSI = LO/HI) and parasympathetic (PNSI = HI/TOTAL) activity were calculated. Power in all bands fell progressively with increasing exercise intensity from rest to trot. At the gallop VLF and LO power continued to fall but HI power rose. SNSI rose from rest to walk, then fell with increasing effort and was lowest at the gallop. PNSI fell from rest to walk, then rose and was highest at the gallop. Normalised HI power exceeded combined VLF and LO power at all gaits, with the ratio HI to LO power being lowest at the walk and highest at the gallop. ANS indicators showed considerable inter-horse variation, and varied less consistently than raw power with increasing physical effort. In the horses studied, the relationship between power and HR changed at exercise intensities associated with heart rates above approximately 120-130 beats/min. At this level, humoral and other non-neural mechanisms may become more important than autonomic modulation in influencing heart rate and heart rate variability (HRV). HRV at intense effort may be influenced by respiratory-gait entrainment, energetics of locomotion and work of breathing. HRV analysis in the frequency domain would appear to be of potential value as a noninvasive means of assessing autonomic modulation of heart rate at low exercise intensities, only. The technique may be a sensitive method for assessing exercise response to experimental manipulations and disease states. PMID- 10836483 TI - An in vitro study to compare 5 different techniques for injection of the navicular bursa in the horse. AB - Numerous different techniques for injection of the navicular bursa have been described but there is little conformity between these descriptions. A literature review was performed and techniques for injection of the navicular bursa were categorised into 5 approaches. This study was performed to evaluate the reproducibility of each technique and to assess which technique was consistently most successful. Five inexperienced operators performed each technique on 5 cadaver forelimbs. Once a needle had been placed, a radiograph was taken to confirm placement was in accordance with the description of the technique. Contrast agent was then injected and another radiograph taken to determine whether the navicular bursa had been injected successfully. The position of the navicular bone was highly predictable as a point 1 cm distal to the coronary band and halfway between the most dorsal and most palmar aspect of the coronary band. The use of this point as a landmark for injection of the navicular bursa allowed a high degree of accuracy and reliability of needle placement, irrespective of foot conformation. PMID- 10836485 TI - Schizotypal traits, attention and eye movements. AB - 1. Subjects demonstrating high, average, or low schizotypal traits participated in saccade tasks of eye movements and attention including: a simple saccade task, an antisaccade task, and/or a cued saccade task measuring both facilitatory effects of cuing and inhibition of return (IOR). 2. Subjects were recruited based on their scores on the Rust Inventory of Schizotypal Cognitions (RISC) and then were given Raine's Schizotypal Personality Questionnaire (SPQ) (1991). 3. Subjects scoring high in schizotypy demonstrated increased errors on the voluntary eye movement task (antisaccade task) (p<0.05). Performance on the reflexive saccade task was not impaired in high compared to low schizotypals, but may have been enhanced as demonstrated by a negative correlation between scores on the SPQ and performance on this task. For the cued saccade task, there were no overall differences in cueing effects between schizotypal groups, however there was a laterality difference between low versus high scoring schizotypal subjects. 4. These results indicate distinct differences in tasks of overt orienting (saccade and antisaccade tasks) and covert orienting tasks (cued saccade task). The patterns of performance by our schizotypy subjects, including impaired voluntary saccade, enhanced reflexive saccade, and lateralized performance on the cued saccade task, are consistent with the performance of schizophrenic patient populations. Thus, our study supports the previous findings of a physiological relationship between schizotypal personality and schizophrenia. PMID- 10836484 TI - Radiographic and clinical survey of degenerative joint disease in the distal tarsal joints in Icelandic horses. AB - The prevalence of degenerative joint disease (DJD) in the distal tarsal joints and the relation between radiographic and clinical signs compatible with the disease were estimated in a population of Icelandic horses used for riding. The material consisted of 614 horses age 6-12 years (mean age = 7.9 years). Radiographs with 3 projections of each tarsus were made and a clinical examination, including palpation of the medial aspect of the distal tarsus and motion evaluation of the hindlimbs before and after a flexion test of the tarsus, was performed. Radiographic signs of DJD in the distal tarsal joints were found in 30.3% of the horses and the prevalence was strongly correlated with age. Hindlimb lameness before and after flexion test and palpation abnormalities were significantly associated with the radiographic findings. The lameness was usually mild and, in most cases, detectable only after the flexion test. The prevalence of lameness was not significantly correlated with age. Lameness could not be predicted by details of the radiographic findings. PMID- 10836486 TI - Clozapine and negative symptoms. An open study. AB - 1. Clozapine, the first atypical antipsychotic, has demonstrated an efficacy in the treatment of resistant schizophrenia. But one of the major challenge in the treatment of schizophrenia remains the lack of efficacy of antipsychotics on negative symptoms of schizophrenia. 2. The authors studied the efficacy of clozapine in an open study in a population of 51 patients, who met the DSM IV criteria for schizophrenia. Using the positive and negative symptom scale (P.A.N.S.S.), and the Extra Pyramidal Symptoms Rating Scale (E.S.R.S.), we try to identify the specificity of the action of clozapine on the different symptomatic dimensions of schizophrenia. 3. The efficacy of clozapine was clinically significant on the negative symptomatology but was delayed compared to the efficacy on the other dimensions of symptomatology evaluated using the PANSS. 4. Nine patients, were considered as deficit patients; in this sample clozapine also demonstrated a significant efficacy on negative symptoms. The efficacy of clozapine did not seem to be a consequence of the better neurological tolerance of this antipsychotic evaluated with ESRS. PMID- 10836487 TI - Pain threshold changes following acupuncture, measured with cutaneous argon laser and electrical tooth pulp stimulation, a comparative study. AB - 1. The present study compared the argon laser technique for pain threshold measurements with the previously often used threshold measurements with electrical tooth pulp (TP) stimulation. 2. Conditioning stimuli with acupuncture was given since it previously has been shown to induce pain threshold changes. Acupuncture needles were inserted in the hands and arms and electrically stimulated. 3. The experiments were performed in a single blind fashion comparing pain threshold effects after either acupuncture or placebo. Pain threshold measurements with a method of limits were used with both electrical TP and the argon laser method. 4. In comparison to placebo the pain thresholds increased after acupuncture, both on the face and on the hand, with the argon laser method, while the TP thresholds were not influenced. No significant pain threshold change was detected with either the argon laser or the TP method following placebo treatment. 5. The results indicate that the argon laser used for sensory testing with a method of limits is a useful method for pain threshold measurements. The difference in pain threshold effects induced by acupuncture measured with the argon laser and the TP stimulation, are discussed and related to mechanisms that may differ between the two types of pain threshold measurements. PMID- 10836488 TI - Pain threshold measurements with cutaneous argon laser, comparing a forced choice and a method of limits. AB - 1. With the argon laser technique cutaneous nociceptors can be activated with high specificity and reproducibility making it a useful tool in psychophysical pain studies. This study was designed to examine and compare two different psychophysical methods combined with the argon laser technique. 2. Pain thresholds on different locations of the body and on different skin types were measured with (i) the method of limits and (ii) the forced choice method. 3. A significant correlation between the pain thresholds measured with the two different methods was detected on feet and hands. The method of limits yielded significantly higher pain thresholds in glabrous skin than hairy skin. Higher pain thresholds were also detected on the right side of the body. No statistically significant difference between sexes was found. A high reproducibility over time as well as in-between investigators was found for the method of limits. 4. Its concluded that the argon laser for pain threshold measurement with the method of limits is useful and preferred to the forced choice, since the method of limits is easier to perform and also less time consuming. PMID- 10836489 TI - 10 microg CCK-4 premedication and 35% CO2 challenge in healthy volunteers. AB - 1. The purpose of the study was to confirm whether a subthreshold dose of CCK-4 would enhance the vulnerability of healthy subjects to a 35% carbon dioxide challenge. 2. 40 subjects, with no prior or present psychiatric disorder and in good physical health were challenged with a vital capacity breath of a 35% carbon dioxide 65% oxygen mixture, immediately after an intravenous injection of 10 microg CCK-4 or placebo, according to a random order, double blind, separate group design. 3. Subjects reported significantly less anxiety and panic symptoms upon carbon dioxide after premedication with CCK-4 than after placebo. 4. CCK-4 and carbon dioxide seem to inhibit rather than enhance each other's effects, possibly through an effect on different neurobiological systems. PMID- 10836490 TI - Functional brain imaging and pharmacotherapy in social phobia: single photon emission computed tomography before and after treatment with the selective serotonin reuptake inhibitor citalopram. AB - 1. Despite increased understanding of the prevalence and pharmacotherapy of social phobia (or social anxiety disorder), the neurobiology of the disorder is little understood. 2. Little data exists on the effect of pharmacotherapeutic intervention on regional cerebral blood flow (rCBF) in this disorder. Patients (n=15) who met DSM-IV diagnostic criteria for social phobia were subjected to single photon emission computed tomography (SPECT) with technetium-99m hexamethylpropylene amine oxime (Tc-99m HMPAO) before and after an eight-week trial of pharmacotherapy with the selective serotonin reuptake inhibitor (SSRI) citalopram. 3. Pharmacotherapy led to significantly reduced activity in the anterior and lateral part of the left temporal cortex; the anterior, lateral and posterior part of the left mid frontal cortex; and the left cingulum. 4. Despite the small sample size, medication non-responders (n=6) had higher activity at baseline in the anterior and lateral part of the left temporal cortex and the lateral part of the left mid frontal regions compared with responders (n=9). These data from this exploratory study are consistent with work suggesting that the anxiety disorders share certain mediating neurocircuitry, although activity in other brain regions may differ. 5. Further research is necessary to determine the neurobiological underpinnings of social phobia. PMID- 10836491 TI - Modulation of estrous cycle and LH, FSH and melatonin levels by pinealectomy and sham-pinealectomy in female rats. AB - 1. The pineal has been shown to have a role in controlling reproduction of polyestrus mammals (like humans and laboratory rodents). It influences the age of sexual maturation; the timing of the ovulatory cycle; and gonadal steroidogenesis. 2. Here the authors report the early and late effects of pinealectomy (Px) and sham-pinealectomy (SPx) on the estrous cycle periodicity, plasma LH, FSH and urinary 6-sulphatoxymelatonin (6-SMT) excretion in female rats. 3. Female Wistar rats (3-4 months of age) were maintained on 12/12 L/D cycle. Orbital venous plexus blood and urine samples were collected from the same rat during the estrus phase before surgery, 4-7 and 55-60 days post surgery. 4. Daily vaginal smears were taken to monitor the estrous cycle and they showed a time dependent increase in the estrus stage duration in Px rats (estrus stage: 1 day in control; 3-4 days after 45 days Px). 5. The decrease of gonadotropins at early post Px was due to surgical stress. 6. 6-SMT levels were significantly lower at 4-7 days post SPx, but at 55-60 d post surgery these levels returned to control values, which indicate pineal gland integrity. The reduction in urinary 6 SMT may be attributed to a possible high level of plasma corticosterone occurring after surgical manipulations. 7. 6-SMT levels in Px rats were extremely lower at 4-7 and 55-60 days post surgery, but not null, confirming the surgical removal of the pineal gland and indicating the synthesis of melatonin in sites other than the pineal gland. PMID- 10836492 TI - Open field is more sensitive than automated activity monitor in documenting ouabain-induced hyperlocomotion in the development of an animal model for bipolar illness. AB - 1. Intracerebroventricular (i.c.v.) administration of ouabain to rats induces motor hyper- and hypoactivity that have been hypothesized to model the mania and depression of bipolar illness, respectively. 2. The extent of ouabain-induced change in activity may vary according to the test environment. 3. To determine the degree of differential response to i.c.v. ouabain in the open field and automated activity monitors, the authors examined a large number of animals (n=40) in both environments. 4. I.c.v. ouabain produced a four-fold increase in open field activity versus i.c.v. artificial cerebrospinal fluid (aCSF) (mean +/- SD: 258.7 +/- 316.61 vs. 84.8 +/- 86.16 squares traversed) (t = 2.648, P = 0.011), but did not alter horizontal activity in automated activity monitors (8193.5 +/- 4902.52 vs. 7088.47 +/- 3046.85 beam interruptions) (t = 0.847, P = 0.4). This increase in activity persisted for at least one week (161.0 +/- 186.35 for i.c.v. ouabain vs. 46.1 +/- 47.46 for i.c.v. aCSF, P = 0.065). 5. Open field is superior to automated activity monitors in capturing ouabain-induced hyperlocomotion response. PMID- 10836493 TI - Effects of 3,4-methylenedioxy-methamphetamine (MDMA) on anxiety in mice tested in the light-dark box. AB - 1. The effects of acute administration of 3,4-methylenedioxymethamphetamine (MDMA; "ecstasy") on anxiety tested in the light/dark box were examined in albino male mice of the OF.1 strain. 2. Animals were evaluated in the light/dark test 30 min after injection of MDMA (1, 8, and 15 mg/kg, i.p) or saline. The following parameters were recorded (for 5 min); (a) number of exploratory rearings in the light and dark sections; (b) number of transitions between the lit and dark areas; (c) time spent in the light and dark areas; (d) latency of the initial movement from the light to the dark area, and (e) locomotor activity in light area. 3. MDMA (8 and 15 mg/kg) produced a significant reduction in exploratory activity (rearings and transitions), without decreasing motility, in comparison with saline-treated mice. However, time spent in lit/dark compartments was not significantly affected by the drug, which could be a consequence of the anti exploratory properties of MDMA. 4. Overall, the behavioral profile found in the light/dark test indicates an anxiogenic-like activity of MDMA in mice. It is suggested, however, that animal models of anxiety which emphasize a social interaction could be more sensitive to the effects of this substance. PMID- 10836494 TI - Female vs. male outpatient depression: a 448-case study in private practice. AB - 1. Aims of the study were to find the prevalence of female depression, and to study the differences between female and male depression, in private practice. Four hundred forty eight consecutive unipolar and bipolar II depressed outpatients were interviewed with the Structured Clinical Interview for DSM-IV, the Montgomery Asberg Depression Rating Scale, and the Global Assessment of Functioning scale. 2. Prevalence of female depression was 67.8%. Female to male ratio was 2:1. Among the variables investigated (age at intake, age at onset, duration of illness, severity, chronicity, psychosis, recurrences, atypical features, comorbidity), atypical features and comorbidity were significantly associated with female depression. 3. Private practice outpatient female depression (chronic, psychotic, or recurrent) was not more severe than male depression. PMID- 10836495 TI - The structural basis of muscle contraction. AB - The myosin cross-bridge exists in two conformations, which differ in the orientation of a long lever arm. Since the lever arm undergoes a 60 degree rotation between the two conformations, which would lead to a displacement of the myosin filament of about 11 nm, the transition between these two states has been associated with the elementary 'power stroke' of muscle. Moreover, this rotation is coupled with changes in the active site (CLOSED to OPEN), which probably enable phosphate release. The transition CLOSED to OPEN appears to be brought about by actin binding. However, kinetics shows that the binding of myosin to actin is a two-step process which affects both ATP and ADP affinity and vice versa. The structural basis of these effects is only partially explained by the presently known conformers of myosin. Therefore, additional states of the myosin cross-bridge should exist. Indeed, cryoelectron microscopy has revealed other angles of the lever arm induced by ADP binding to a smooth muscle actin-myosin complex. PMID- 10836496 TI - Mechanics and models of the myosin motor. AB - In striated muscles, shortening comes about by the sliding movement of thick filaments, composed mostly of myosin, relative to thin filaments, composed mostly of actin. This is brought about by cyclic action of 'cross-bridges' composed of the heads of myosin molecules projecting from a thick filament, which attach to an adjacent thin filament, exert force for a limited time and detach, and then repeat this cycle further along the filament. The requisite energy is provided by the hydrolysis of a molecule of adenosine triphosphate to the diphosphate and inorganic phosphate, the steps of this reaction being coupled to mechanical events within the cross-bridge. The nature of these events is discussed. There is good evidence that one of them is a change in the angle of tilt of a 'lever arm' relative to the 'catalytic domain' of the myosin head which binds to the actin filament. It is suggested here that this event is superposed on a slower, temperature-sensitive change in the orientation of the catalytic domain on the actin filament. Many uncertainties remain. PMID- 10836497 TI - Single-motor mechanics and models of the myosin motor. AB - Recent progress in single-molecule detection techniques is remarkable. These techniques have allowed the accurate determination of myosin-head-induced displacements and how mechanical cycles are coupled to ATP hydrolysis, by measuring individual mechanical events and chemical events of actomyosin directly at the single-molecule level. Here we review our recent work in which we have made detailed measurements of myosin step size and mechanochemical coupling, and propose a model of the myosin motor. PMID- 10836498 TI - Searching for kinesin's mechanical amplifier. AB - Kinesin, a microtubule-based motor, and myosin, an actin-based motor, share a similar core structure, indicating that they arose from a common ancestor. However, kinesin lacks the long lever-arm domain that is believed to drive the myosin power stroke. Here, we present evidence that a much smaller region of ca. 10-40 amino acids serves as a mechanical element for kinesin motor proteins. These 'neck regions' are class conserved and have distinct structures in plus-end and minus-end-directed kinesin motors. Mutagenesis studies also indicate that the neck regions are involved in coupling ATP hydrolysis and energy into directional motion along the microtubule. We suggest that the kinesin necks drive motion by undergoing a conformational change in which they detach and re-dock onto the catalytic core during the ATPase cycle. Thus, kinesin and myosin have evolved unique mechanical elements that amplify small, nucleotide-dependent conformational changes that occur in their similar catalytic cores. PMID- 10836499 TI - The conformational cycle of kinesin. AB - The stepping mechanism of kinesin can be thought of as a programme of conformational changes. We briefly review protein chemical, electron microscopic and transient kinetic evidence for conformational changes, and working from this evidence, outline a model for the mechanism. In the model, both kinesin heads initially trap Mg x ADP. Microtubule binding releases ADP from one head only (the trailing head). Subsequent ATP binding and hydrolysis by the trailing head progressively accelerate attachment of the leading head, by positioning it closer to its next site. Once attached, the leading head releases its ADP and exerts a sustained pull on the trailing head. The rate of closure of the molecular gate which traps ADP on the trailing head governs its detachment rate. A speculative but crucial coordinating feature is that this rate is strain sensitive, slowing down under negative strain and accelerating under positive strain. PMID- 10836500 TI - Structural model of F1-ATPase and the implications for rotary catalysis. AB - The crystal structure of bovine mitochondrial F1-ATPase is described. Several features of the structure are consistent with the binding change mechanism of catalysis, in which binding of substrates induces conformational changes that result in a high degree of cooperativity between the three catalytic sites. Furthermore, the structure also suggests that catalysis is accompanied by a physical rotation of the centrally placed gamma-subunit relative to the approximately spherical alpha3beta3 subassembly. PMID- 10836501 TI - A rotary molecular motor that can work at near 100% efficiency. AB - A single molecule of F1-ATPase is by itself a rotary motor in which a central gamma-subunit rotates against a surrounding cylinder made of alpha3beta3 subunits. Driven by the three betas that sequentially hydrolyse ATP, the motor rotates in discrete 120 degree steps, as demonstrated in video images of the movement of an actin filament bound, as a marker, to the central gamma-subunit. Over a broad range of load (hydrodynamic friction against the rotating actin filament) and speed, the F1 motor produces a constant torque of ca. 40 pN nm. The work done in a 120 degree step, or the work per ATP molecule, is thus ca. 80 pN nm. In cells, the free energy of ATP hydrolysis is ca. 90 pN nm per ATP molecule, suggesting that the F1 motor can work at near 100% efficiency. We confirmed in vitro that F1 indeed does ca. 80 pN nm of work under the condition where the free energy per ATP is 90 pN nm. The high efficiency may be related to the fully reversible nature of the F1 motor: the ATP synthase, of which F1 is a part, is considered to synthesize ATP from ADP and phosphate by reverse rotation of the F1 motor. Possible mechanisms of F1 rotation are discussed. PMID- 10836503 TI - Theories of rotary motors. AB - The bacterial flagellar motor and the ATP-hydrolysing F1 portion of the F1Fo ATPase are known to be rotary motors, and it seems highly probable that the H+ translocating Fo portion rotates too. The energy source in the case of Fo and the flagellar motor is the flow of ions, either H+ (protons) or Na+, down an electrochemical gradient across a membrane. The fact that ions flow in a particular direction through a well-defined structure in these motors invites the possibility of a type of mechanism based on geometric constraints between the rotor position and the paths of ions flowing through the motor. The two best studied examples of such a mechanism are the 'turnstile' model of Khan and Berg and the 'proton turbine' model of Lauger or Berry. Models such as these are typically represented by a small number of kinetic states and certain allowed transitions between them. This allows the calculation of predictions of motor behaviour and establishes a dialogue between models and experimental results. In the near future structural data and observations of single-molecule events should help to determine the nature of the mechanism of rotary motors, while motor models must be developed that can adequately explain the measured relationships between torque and speed in the flagellar motor. PMID- 10836502 TI - Constraints on models for the flagellar rotary motor. AB - Most bacteria that swim are propelled by flagellar filaments, each driven at its base by a rotary motor embedded in the cell wall and cytoplasmic membrane. A motor is about 45 nm in diameter and made up of about 20 different kinds of parts. It is assembled from the inside out. It is powered by a proton (or in some species, a sodium-ion) flux. It steps at least 400 times per revolution. At low speeds and high torques, about 1000 protons are required per revolution, speed is proportional to protonmotive force, and torque varies little with temperature or hydrogen isotope. At high speeds and low torques, torque increases with temperature and is sensitive to hydrogen isotope. At room temperature, torque varies remarkably little with speed from about -100 Hz (the present limit of measurement) to about 200 Hz, and then it declines rapidly reaching zero at about 300 Hz. These are facts that motor models should explain. None of the existing models for the flagellar rotary motor completely do so. PMID- 10836504 TI - The role of thermal activation in motion and force generation by molecular motors. AB - The currently accepted mechanism for ATP-driven motion of kinesin is called the hand-over-hand model, where some chemical transition during the ATP hydrolysis cycle stretches a spring, and motion and force production result from the subsequent relaxation. It is essential in this mechanism for the moving head of kinesin to dissociate, while the other head remains firmly attached to the microtubule. Here we propose an alternative Brownian motor model where the action of ATP modulates the interaction potential between kinesin and the microtubule rather than a spring internal to the kinesin molecule alone. In this model neither head need dissociate (which predicts that under some circumstances a single-headed kinesin can display processive motion) and the transitions by which the motor moves are best described as thermally activated steps. This model is consistent with a wide range of experimental data on the force-velocity curves, the one ATP to one-step stoichiometry observed at small load, and the stochastic properties of the stepping. PMID- 10836505 TI - How Fo-ATPase generates rotary torque. AB - The F-ATPases synthesize ATP using a transmembrane ionmotive force (IMF) established by the electron transport chain. This transduction involves first converting the IMF to a rotary torque in the transmembrane Fo portion. This torque is communicated from Fo to the F1 portion where the energy is used to release the newly synthesized ATP from the catalytic sites according to Boyer's binding change mechanism. Here we explain the principle by which an IMF generates this rotary torque in the Fo ion engine. PMID- 10836506 TI - Cooperativity of myosin molecules through strain-dependent chemistry. AB - There is mounting evidence that the myosin head domain contains a lever arm which amplifies small structural changes that occur at the nucleotide-binding site. The mechanical work associated with movement of the lever affects the rates at which the products of ATP hydrolysis are released. During muscle contraction, this strain-dependent chemistry leads to cooperativity of the myosin molecules within a thick filament. Two aspects of cooperative action are discussed, in the context of a simple stochastic model. (i) A modest motion of the lever arm on ADP release can serve to regulate the fraction of myosin bound to the thin filament, in order to recruit more heads at higher loads. (ii) If the lever swings through a large angle when phosphate is released, the chemical cycles of the myosin molecules can be synchronized at high loads. This leads to stepwise sliding of the filaments and suggests that the isometric condition is not a steady state. PMID- 10836507 TI - Past, present and future experiments on muscle. AB - Since the basic outline of the sliding filament mechanism became apparent some 45 years ago, the principal challenge, an experimental one, has been to produce definitive evidence about the detailed molecular mechanisms by which myosin cross bridges produce force and movement in a muscle. More recently, similar questions could be posed about other molecular motors, in non-muscle cells. This problem proved unexpectedly difficult to solve, in part because of the technical difficulty of obtaining the structural and mechanical information required about rapid events within macromolecules, especially in a working system, and this triggered many remarkable technical developments. There is now very strong evidence for a large change in shape of the myosin heads during ATP hydrolysis, consistent with a lever-arm mechanism. Whether this does indeed provide the driving force for contraction and movement--and, if so, exactly how--and whether some other processes could also play a significant role, is discussed in the light of the experimental and theoretical findings presented at this meeting, and other recent and long-term evidence. PMID- 10836508 TI - Epitope-specific impairment of production of antibody against merozoite surface glycoprotein 1 of Plasmodium falciparum in symptomatic patients with malaria. AB - We analyzed the relationships between levels of antibody specific for merozoite surface glycoprotein-1 (MSP1) of Plasmodium falciparum and clinical manifestations in humans. We prepared recombinant MSP1 proteins representing block 3 (M3), block 6 (M6), blocks 1-6 (M1/6), and block 17. When we divided the slide-positive individuals in Guadalcanal into symptomatic and asymptomatic groups, the former group showed lower IgG levels against M6 and block 17, but not against M3, than did the asymptomatic group (P < 0.01). The possibility of nonspecific suppression was unlikely, given that the levels of antibody against poliomyelitis virus observed in the two groups were almost the same. Among the IgG subclasses tested, production of cytophilic IgG3 seemed to be dominant. When we analyzed epitopes recognized by antibodies against block 17, a peptide (SSSNFLGIS) was preferentially recognized by sera from asymptomatic individuals. These results suggest that clinical symptoms occurring during falciparum malaria seem to be associated with the development of levels of antibody against particular epitopes on MSP1, which is under the control of an immunoregulatory mechanism. PMID- 10836509 TI - Ribosomal small-subunit RNA gene-sequence analysis of Theileria lestoquardi and a Theileria species highly pathogenic for small ruminants in China. AB - A fatal disease of sheep and goats in the northwestern part of China has been reported to be due to Theileria lestoquardi (syn. T. hirci). However, some characteristics of the causative agent are not in accordance with attributes ascribed to this parasite. We therefore determined the nucleotide sequence of the small-subunit ribosomal RNA (srRNA) gene of T. lestoquardi and the parasite identified in China and compared it with that of other Theileria and Babesia species. In the inferred phylogenetic tree the srRNA sequence of the Chinese parasite was found to be most closely related to T. buffeli and clearly divergent from T. lestoquardi, suggesting that it is an as yet unrecognized Theileria species. Extensive structural similarities were observed between the srRNA sequences of T. lestoquardi and T. annulata, revealing a close phylogenetic relationship between these two Theileria species. On the basis of the srRNA nucleotide sequence, polymerase chain reaction (PCR) primers were designed that specifically amplified genomic DNA of the Chinese Theileria species. These primers may be valuable tools in future epidemiology studies. PMID- 10836510 TI - Parasite communities of the Salzhaff (Northwest Mecklenburg, Baltic Sea) II. Guild communities, with special regard to snails, benthic crustaceans, and small sized fish. AB - Metazoan parasites of guilds of benthic snails and crustaceans and of four fish families--Gobiidae, Gasterosteidae, Syngnathidae, and Zoarcidae--were investigated off the brackish Salzhaff area (Southwest Baltic) in the semienclosed Salzhaff and the near Rerik Riff in the free Baltic. Comparisons revealed greater similarities in parasite populations and communities within the fish guilds than between them. According to an evaluation of the core-/satellite species concept using abundance values, the most important parasites of fish were some generalists, such as Cryptocotyle spp., Podocotyle atomon, and Diplostomum spathaceum, as well as some specialists, such as Acanthostomum balthicum, Thersitina gasterostei, and Aphalloides timmi. These specialists revealed high degrees of prevalence in their main hosts and lower degrees in one or two by hosts. Additional importance is assigned to parasites that cause harm to their hosts due to their large size, e.g., Schistocephalus spp., or via massive infestation, e.g., several digenean metacercariae. Because specialists were more prominent in snails and fish from the Rerik Riff, the correlation of host numbers with prevalence resulted in only a slight increase instead of a more rapid rise in regression among crustaceans and fish from the entire Salzhaff, where the generalists were more prevalent. The selected host guilds demonstrated the entire life cycles of three digeneans (P. atomon, A. balthicum, A. timmi), one acanthocephalan (Echinorhynchus gadi), and one nematode (Hysterothylacium sp.). The prevalence increased in these cycles from host level to host level and attained relatively high values in all guilds. The parasite fauna of the Salzhaff area is influenced by eutrophication stress, which leads to a high level of productivity and, consequently, to great densities in primary consumers such as snails and crustaceans. These are attractive for several secondary consumers such as fish and birds, which is the reason for the existence of at least 24 autogenic and 20 allogenic parasite species at this locality. The slight surplus of the first category indicates a yet-balanced environment in the investigation area. PMID- 10836511 TI - The fine structure of acidocalcisomes in Trypanosoma cruzi. AB - Trypanosoma cruzi survives in vertebrate and invertebrate hosts and has developed mechanisms that allow it to adapt to changes in the microenvironment such as temperature, pH, and ionic composition. Most of its calcium is concentrated in an organelle named the acidocalcisome, which is acidified by a (V-H+)-adenosine triphosphatase and has H+/Ca2+ counter-transportation for calcium uptake. In this work, acidocalcisomes were examined using different transmission electron microscopy techniques. In thin sections of different stages, acidocalcisomes presented a circular shape with an electron-dense inclusion containing P3-, Ca2+, Na+, Mg2+, K+, and Zn2+. They could be distinguished from gold-labeled albumin containing reservosomes in whole epimastigotes, and a morphometric analysis showed higher amounts of these organelles in amastigotes as compared with epimastigotes and trypomastigotes. It is possible that this variation in the amount of acidocalcisomes in the different evolutive stages could reflect adaptation mechanisms used by the parasite to survive and multiply in different environmental conditions. PMID- 10836512 TI - Investigation of different ontogenetic stages of Raillietiella sp. (Pentastomida: Cephalobaenida): suboral gland and frontal gland. AB - In Raillietiella sp. larvae we discovered a gland whose duct pores shed their secretion caudally of the mouth onto the integument. This gland type, which we call the suboral gland, seems to be characteristic of all pentastomid species. We discuss its possible function in the invasion of the larvae into the intermediate host. Furthermore, we identified frontal gland cells and their ducts in Raillietiella and differentiated them from buccal glands and their ducts. They appear to be a general feature of pentastomids and exist even in early ontogenetic stages. PMID- 10836513 TI - Infectivity, predilection sites, and freeze tolerance of Trichinella spp. in experimentally infected sheep. AB - A total of 36 sheep in groups of 4 were inoculated with 9 isolates of Trichinella and euthanized after 10 weeks. Thereafter, numbers of muscle larvae were determined in 13 different muscles/muscle groups. Muscle larvae were found in high numbers in all four sheep inoculated with T. spiralis, in lower numbers in two sheep inoculated with T. pseudospiralis (USA isolate), and in very low numbers in one sheep inoculated with T. pseudospiralis (USSR isolate) and one inoculated with T. britovi. In infections of high and moderate larval intensity, predilection sites of T. spiralis were the masseter muscles, the tongue, and the diaphragm and those of T. pseudospiralis were the masseter muscle and the neck. In low-intensity infections, muscle larvae were detected only in the diaphragm or in pooled muscle samples. For evaluation of the freeze tolerance of the different Trichinella species in sheep-muscle tissue, samples taken from the filet were stored at +5 degrees, -5 degrees, and -18 degrees C, respectively. After exposure for 1 and 4 weeks the tissue was digested and the released larvae were inoculated into mice for determination of the reproductive capacity index (RCI). Larvae of both T. spiralis and T. pseudospiralis survived freezing at -5 degrees and -18 degrees C for 4 weeks. PMID- 10836514 TI - Involvement of tumor necrosis factor-alpha during infection of human monocytic cells by Toxoplasma gondii. AB - Although the involvement of tumor necrosis factor-alpha (TNF-alpha) during Toxoplasma gondii infection has largely been described in mouse models, only a few studies in human models have been reported. We demonstrated the role of TNF alpha during the process of invasion by T. gondii in human monocytic cells. Cotreatment of cells with interferon-gamma (IFN-gamma) and lipolysaccharide (LPS) during T. gondii infection induced TNF-alpha production and decreased the number of parasitized cells (invasion index). Neutralization of the production of TNF alpha using a specific monoclonal antibody or pentoxifylline enhanced the invasion index. The relationship between TNF-alpha production and protection of monocytic cells against T. gondii invasion was confirmed by treatment of infected cultures with exogenous TNF-alpha. Thus, in contrast to the results obtained in murine models but in accordance with those observed in other human models, the present study shows for the first time in human monocytic cells that T. gondii does not induce any TNF-alpha secretion and inhibits TNF-alpha production induced by IFN-gamma/LPS. PMID- 10836515 TI - Low temperature blocks fluid-phase pinocytosis and receptor-mediated endocytosis in Trypanosoma cruzi epimastigotes. AB - Gold-labeled albumin and transferrin were used to follow at the ultrastructural level the early events and the effect of low temperature on protein uptake by Trypanosoma cruzi epimastigotes. In parasites incubated for 5 min at 28 degrees C with protein-gold complexes, extracellular markers were found only at the cytostome and/or the flagellar pocket regions, whereas intracellular gold particles were detected inside small uncoated vesicles located nearby. Within 10 min, labeling was also observed in uncoated vesicles close to the nucleus. Only after 30 min could the tracers be detected in the reservosomes. Weak labeling in the cytostome and flagellar pocket of parasites incubated at 4 degrees C with the albumin-gold solution indicated that albumin uptake occurred by fluid-phase pinocytosis. On the other hand, intense labeling at the cytostome was observed in parasites incubated at 4 degrees C with gold-labeled transferrin, showing that receptor-mediated endocytosis occurs mainly at this site. Both proteins were absent from the cells at 4 degrees C and 12 degrees C. Raising the temperature from 12 degrees C to 28 degrees C led to transferrin labeling in intracellular vesicles dispersed throughout the cytoplasm, but not in reservosomes. Our results suggest that low temperatures affect the transport and pinching of endocytic vesicles as well as the rate of delivery of transferrin to reservosomes. PMID- 10836516 TI - Allozyme analysis of two polymorphic enzymes in a natural population of Lecithochirium musculus. AB - Genetic variability among 112 individuals of Lecithochirium musculus from Anguilla anguilla from northwestern Spain was examined using allozyme analysis. Starch-gel electrophoresis was used to investigate the genetic variation in two polymorphic enzymes, glucose phosphate isomerase (GPI) and phosphoglucomutase (PGM). The banding patterns obtained for GPI were consistent with a dimeric structure for this enzyme and with single-locus control. The inferred genotype frequencies did not significantly differ as compared with those predicted by the Hardy-Weinberg equation. Two PGM loci were detected (Pgm-1 and Pgm-2). Pgm-1 was polymorphic and showed a striking departure from Hardy-Weinberg predictions (total absence of heterozygotes for alleles in high frequencies), raising the possibility of the existence of two reproductively isolated populations. However, other reasons for this observation are discussed. PMID- 10836517 TI - Immunomodulation of lambs following treatment with a proteasome preparation from infective larvae of Trichostrongylus colubriformis. AB - Proteinases are known to be capable of prolonging the survival of endoparasites in a host. We were therefore interested in knowing whether immunization of lambs against a proteasome (multisubunit proteinases) preparation obtained from Trichostrongylus colubriformis infective third-stage larvae (L3) would have any effect on the immune response to a single challenge infection with the same organism. A total of 21 penned lambs aged 8 months were divided into 3 equal groups. Group 1 was immunized on three occasions with increasing amounts of a proteasome-enriched fraction obtained from infective L3. Group 2 was given a similar amount of protein from the initial supernatant of homogenized larvae. Group 3 (controls) received adjuvant plus saline solution only. All groups were challenged with 60,000 infective T. colubriformis larvae at 28 days after the last immunization. Significant protection was obtained only when the initial supernatant extract was used to immunize lambs. The proteasome preparation seemed to have immunosuppressive effects through the stimulation of nonspecific IgE production. Significantly lower levels of specific IgE were observed in lambs immunized with the proteasome-enriched fraction, and levels of specific IgG antibodies were increased. We suggest that proteasome fractions of T. colubriformis may serve as useful preparations for the study of mechanisms of IgE production in parasitized sheep. PMID- 10836518 TI - Comparative ultrastructure of eggs in Echinostoma paraensei, E. caproni, and E. trivolvis (Trematoda: Echinostomatidae). AB - The 37-collar-spined echinostomes Echinostoma paraensei, E. caproni, and E. trivolvis are digeneans that live in the intestine of small mammals and birds. Comparative studies of the eggs of these species were done using light microscopy (LM), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). The egg of E. caproni was the largest of the three species studied, whereas the egg of E. trivolvis was the smallest in both length and width. The SEM study showed differences in the aboperculum region of the eggs in all three species. The TEM study showed that the eggshell of all three species consisted of three layers, but no difference in eggshell structure was observed in any species. PMID- 10836519 TI - The gene encoding the metacyclogenesis-associated transcript Mat-1 is conserved in the genus Leishmania and shows a tendency to form dimers upon protein expression. AB - The Leishmania infantum Mat-1 gene--recently described in L. major as a highly stage-specific, metacyclogenesis-associated transcript--has been cloned. The 420 bp Mat-1 coding region is conserved with respect to the L. major gene (82% sequence homology). Analysis of the predicted amino-acid sequence reveals structural motifs showing homology with the class of leucine-zipper transcription factors. Southern-blot hybridization analysis suggests that Mat-1 is a low-copy number gene, probably consisting of two gene copies. The recombinant Mat-1 protein expressed in fusion with the Escherichia coli maltose-binding protein shows a tendency to form dimers in the presence of the leucine-rich C-terminal domain. Bacteria expressing the Mat-1 open reading frame are highly growth attenuated and tend to delete or modify the insert, which suggests that expression of Mat-1 is toxic for the bacteria. PMID- 10836520 TI - Serum amyloid A--an indicator of inflammation in ankylosing spondylitis. AB - Biological markers of inflammation are useful for the diagnosis and monitoring of inflammatory rheumatic diseases. The present study tested, whether serum amyloid A (SAA) could be used as a marker of inflammatory disease activity in ankylosing spondylitis (AS). In 72 patients with AS, the two valuable surrogate markers of disease activity, erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP), and an established clinical activity score (Bath Ankylosing Spondylitis Disease Activity Index, BASDAI) were correlated to the serum levels of SAA. It was found that SAA correlates well with ESR, CRP, and BASDAI. Because of its strong correlation, SAA seems to be an additional very useful disease activity marker. When used in diagnosis, and especially in monitoring of inflammation, further studies are required. Another interesting point of view is the described role of plasma SAA as a precursor of Amyloid A (AA) protein in secondary amyloidosis, a known complication in AS. In all probability, high circulating SAA levels are a predisposing indicator of disease activity. PMID- 10836521 TI - Basic FGF-induced activation of telomerase in rheumatoid synoviocytes. AB - To investigate the mechanism of persistent proliferation of rheumatoid arthritis (RA) synoviocytes in situ, we examined the activity of telomerase enzyme and the expression of telomerase related factors in cultured synoviocytes. Cultured synoviocytes obtained from patients with rheumatoid arthritis (n = 29), osteoarthritis (OA, n = 18), and traumatic joint disease (TJD, n = 4) were examined. Telomerase activity was detected by TRAP (telomeric repeat amplification protocol) assay, and 12 out of 29 samples of synoviocytes (41%) from RA patients showed a positive telomerase activity, whereas none of the samples from OA and TJD patients showed this activity. Results were confirmed by PCR-ELISA. The telomerase activity was enhanced by basic fibroblast growth factor (bFGF). The mRNA expression of telomerase related factors, such as hTERC, TRF2, and TEP-1, showed no difference between RA and OA synoviocytes. Our results suggest that telomerase is activated in rheumatoid synoviocytes, and that bFGF upregulates the activity of this enzyme in RA synoviocytes. PMID- 10836522 TI - Clinical associations and characterisation of antineutrophil cytoplasmic antibodies directed against bactericidal/permeability-increasing protein and azurocidin. AB - Bactericidal/permeability-increasing protein (BPI) and azurocidin (AZ) are recently described target antigens of antineutrophil cytoplasmic antibodies (ANCA). In this study, BPI-ANCA were demonstrated most often in patients with ulcerative colitis (36/92, 39%), Crohn's disease (17/66, 26%) and cystic fibrosis (11/14, 79%), but also in patients with rheumatoid arthritis (8/40, 20%), systemic lupus erythematosus (SLE) (111/65, 17%) and mixed connective tissue disease (4/18, 22%). BPI-ANCA were also common in sera containing antinuclear (ANA) (9/43, 21%) or antidouble-stranded (ds) DNA (7/28, 25%) antibodies. There was no increased frequency of abnormal alpha1-antitrypsin (alphal1AT) phenotypes in patients with BPI-ANCA, and BPI-ANCA were not more common in individuals with an abnormal phenotype. The predominant IgG subclasses were IgG1 and IgG3; IgA but not IgM was present. Both IgG and IgA BPI-ANCA were high affinity antibodies, and the affinity of IgG antibodies did not change with time in the sera tested. Four of the five sera (80%) containing BPI-ANCA did not bind to denatured, reduced BPI, suggesting that most BPI-ANCA recognised conformational epitopes. AZ-ANCA were demonstrated in 2/11 patients (18%) with Wegener's granulomatosis, 3/12 (25%) with cystic fibrosis and 3/14 (21%) with chronic active hepatitis. AZ-ANCA were present in 5/25 sera (25%) with ANA, but the levels were only marginally elevated. AZ-ANCA were uncommon in patients with inflammatory bowel and rheumatological diseases, and in sera containing other autoantibodies. Again, there was no association with abnormal alpha1-AT phenotypes. BPI represents a major ANCA target antigen in patients with rheumatological as well as inflammatory bowel disease and cystic fibrosis, but AZ-ANCA are uncommon. PMID- 10836523 TI - TNF-alpha gene polymorphism does not affect the clinical and radiological outcome of rheumatoid arthritis. AB - The present study was undertaken in order to investigate the relationship between tumor necrosis factor-alpha (TNF-alpha) gene polymorphism and the radiological progression of rheumatoid arthritis (RA) within the first 3-years of the disease. Sixty-eight RA patients (59 women and nine men) were observed for 3-years. TNF alpha polymorphism analysis was performed in all patients. Radiographs of the hands were taken at the onset of study and after 3-years of follow-up. Radiographs were assessed according to the Larsen index (damage score and progression of damage score). We did not observe any correlation between TNF gene polymorphism and damage score or progression of damage score. The obtained data suggests that TNF-308 polymorphism cannot serve as an indicator of the disease course in RA patients. PMID- 10836524 TI - Mast cell derangement in salivary glands in patients with Sjogren's syndrome. AB - Mast cells (MCs) have been implicated in many immune-inflammatory disorders. Deranged mast cell distribution and function may contribute to the local pathomechanisms in the labial salivary glands (LSG) in Sjogren's syndrome (SS). Evidence for MC presence, localization, frequency, subtype, and degree of activation was sought by using reverse transcriptase-polymerase chain reaction (RT-PCR), immunohistochemistry (IHC)/image analysis, transmission electron microscopy, Western blotting, spectrophotometric activity assay, and radioimmunoassay. The overall expression (densitometric units) of MC tryptase mRNA (1483 +/- 228 vs 1044 +/- 308) did not differ between SS and control LSGs. However, IHC disclosed an uneven distribution of MCs in SS with an absence in lymphocyte foci and increased numbers (cells/mm2 77 +/- 7 vs 38 +/- 4, P < 0.01) elsewhere. Absence of MCs in the lymphocyte foci was not a fixation artefact and was not explained by the presence of "phantom" MCs in these areas. In both SS and controls, 80% of all MCs were chymase containing, but the typical lattices/gratings characteristic for connective tissue MCs (CTMCs) were not found. Instead, MC granules had mostly a homogeneous, finely granular substructure characteristic of "new" granules subjected to a continuous, low grade release. 32 kDa MC tryptase was found in saliva and its activity/concentration was comparable to that found in controls. However, tryptase output was low in SS (1.30 +/- 0.30 microg/min vs 3.49 +/- 0.66 microg/min, P < 0.001). Normal LSGs contain mostly CTMCs, in close contact with various resident and immigrant cells, characterized by a low-grade release of MC mediators. In SS this normal pattern is disturbed so that MCs are absent in lymphocyte foci (but increased elsewhere in the glands). The net salivary output of MC mediators is low in SS. This derangement of MCs may contribute to the pathogenesis of SS via multiple mechanisms. PMID- 10836525 TI - Interleukin-6 serum concentration in ankylosing spondylitis: a reliable predictor of disease progression in the subsequent year? AB - The aim of the present study was to evaluate whether in ankylosing spondylitis (AS), interleukin-6 (IL-6) is a reliable predictor of changes in mobility in the subsequent year. Of 261 AS patients who had been enrolled in a previous study, 128 returned for treatment at our health centre after 1 year (+/-3 months). The variables for mobility after 1 year (II) were compared with the findings of the previous year (I). Differences in parameters for mobility were related to the serum concentration of IL-6 in the previous year. Relation between serum concentration of IL-6 and difference (II-I) in occiput-to-wall distance (Spearman's rank correlation coefficient r(s), P value) was 0.02, 0.82; chin chest distance -0.09, 0.31; cervical rotation -0.08, 0.39; chest expansion 0.05, 0.54; finger-floor distance -0.02, 0.84; Ott sign (flexibility of the thoracic spine) -0.11, 0.22; Schober sign 0.01, 0.94. After 1 year there was a significant improvement in cervical rotation in patients with low IL-6 serum concentration (lower quartile), but not in those with high levels of IL-6 (upper quartile). No further difference was seen between patients with high or low levels of IL-6. The present data suggest that the serum concentration of IL-6 does not allow a prediction of disease progression in the subsequent year. PMID- 10836526 TI - Bone marrow CD34+ progenitor cells from rheumatoid arthritis patients support spontaneous transformation of peripheral blood B cells from healthy individuals. AB - We show that bone marrow (BM) CD34+ progenitor cells from rheumatoid arthritis (RA) patients have the capacity to support spontaneous transformation of peripheral blood B cells. CD34+ cells purified from BM blood from eight RA patients and eight osteoarthritis (OA) patients were expanded with granulocyte/macrophage colony stimulating factor (GM-CSF) for 4-6 weeks. GM-CSF stimulated BM CD34+ cells from three of eight RA patients, but none from seven OA patients, gave rise to spontaneous transformation of highly purified B cells of Epstein-Barr virus (EBV)-seronegative healthy donors. GM-CSF-stimulated BM CD34+ cells from four of six RA patients and from one of four OA patients also supported the spontaneous transformation of peripheral blood B cells from EBV seropositive healthy donors. All the transformed B cell lines were positive for EBV-DNA as determined by PCR. Neither GM-CSF-stimulated BM CD34+ cells alone nor highly purified B cells alone gave rise to spontaneously transformed B cell lines. These results suggest that the capacity of BM CD34+ cells to support survival of B cells might contribute to the pathogenesis of RA by sustaining abnormal B cell responses. PMID- 10836527 TI - Trace element status (Se, Cu, Zn) and serum lipid profile in Portuguese subjects of San Miguel Island from Azores'archipelago. AB - Data on selenium, copper and zinc in serum of Portuguese inhabitants of the city of Ponta Delgada-Azores Archipelago are reported for the first time. The subjects are of both sexes, aged 20 to 60 years, and non-abusers of alcohol, tobacco or drugs. Serum concentrations of these elements are in the same range than those found for populations of Lisbon and of some other European countries. Differences between sexes are observed, with serum selenium and zinc levels being higher in males than in females, and the copper levels being higher in females as compared to males. These results can be explained by the hormonal status and/or oral contraceptive steroids intake, particularly for copper. Concerning lipid profile, the majority of individuals have serum lipid parameters within the normal range. In addition, no difference in trace element levels between normo and hyperlipidemic individuals is observed and no conclusive results about the relationship of all evaluated parameters to alcohol, tobacco and drug consumption are observed, in agreement with data obtained in the population of Lisbon. PMID- 10836528 TI - Assessment of reference values for elements in human hair of urban schoolboys. AB - Hair samples of youngsters (3-15 years of age) from several urban areas of Rome were analyzed to determine the content of 19 minor and trace elements with the aim of assessing Reference Values (RVs). Thirteen essential elements were taken into account, Ca, Co, Cr, Cu, Fe, Mg, Mn, Mo, Ni, P, Se, V and Zn. On the other hand, Al, As, Cd, Pb, Sr and Ti were also evaluated on the basis of their potential toxicity. Procedures were developed for the collection, storage and pre analytical treatment of samples. Measurements were performed by inductively coupled plasma atomic emission spectrometry. Subgroups were formed according to age and sex. Significant differences were found for certain elements depending on age and sex. This was the case, e.g., for Ca which showed a mean value of 336 mg/kg for males and of 537 mg/kg for females. The sex-dependent pattern for this element was also apparent when the three age subgroups of 3-6, 7-10 and 11-15 years were compared. The overall RVs obtained (mg/kg) are as follows Al, 10.2; As, 0.09; Ca, 450; Cd, 0.23; Co, 0.67; Cr, 0.99; Cu, 22.1; Fe, 19.0; Mg, 28.0; Mn, 0.35; Mo, 0.43; Ni, 1.49; P, 195; Pb, 7.11; Se, 0.77; Sr, 1.20; Ti, 0.79; V, 1.22; and Zn 150. PMID- 10836529 TI - Cytotoxic aldehyde generation in heart following acute iron-loading. AB - Although the mechanism of myocardial failure following acute iron poisoning is not known, excess iron-catalyzed free radical generation is conjectured to play a role. The effects of time (0 to 360 minutes) on total iron concentrations, glutathione peroxidase activity, and cytotoxic aldehyde production in heart of mice (B6D2F1, n = 65) were first investigated following acute iron-loading (20 mg iron dextran i.p./mouse). In a subsequent experiment, the effects of dose (0 to 80 mg iron dextran i.p./mouse, n = 75) on the aforementioned parameters were investigated. Our results show that the concentrations of cytotoxic aldehydes: (1) significantly differ over-time, with corresponding increases in total concentrations of iron (r = 0.93, p < 0.001); and (2) increase parallel to the total dose of iron administered (r = 0.95, p < 0.001). Furthermore, dose-and time dependent alterations to glutathione peroxidase activity are observed, which is most likely due to an acute up-regulation of the enzyme as an endogenous protective response to increased free radical activity in the heart subsequent to iron-loading. While no single mechanism is likely to account for the complex pathophysiology of acute iron-induced heart failure, our results shown that iron loading can result in significant free radical generation, as quantified by cytotoxic aldehydes, in heart tissue of mice. This is the first report on the effects of time and dose on cytotoxic aldehyde generation and glutathione peroxidase activity in heart of mice following acute iron-loading. PMID- 10836530 TI - Trace elements in diabetes mellitus. Peculiarities and clinical validity of determinations in blood cells. AB - Significantly more information about trace element status can be obtained by investigating concentrations in blood cells instead of only evaluating the concentrations in plasma. This can be explained by the fact that essential trace elements such as zinc, copper, chromium and selenium take part in a variety of enzymatic processes on a molecular cellular level. Ignoring these important biochemical roles, trace element concentrations determined in whole blood or plasma very often lead to conclusions contrary to the actual intracellular concentration. Especially in metabolic diseases like diabetes mellitus, conclusions drawn from trace element concentrations in blood cells usually offer more valuable clinical information about the metabolic state than trace element concentrations in plasma or whole blood. In the present investigation copper and zinc concentrations were increased in all blood fractions of diabetic patients (IDDM). In insulin-dependent diabetic children significantly higher values of zinc in erythrocytes were also found, and they were higher in patients with poor metabolic control (HbA1c>9%). When different blood fractions in diabetic patients (NIDDM) were compared with a control group, chromium was significantly increased in plasma and polymorphonuclear cells. Patients with IDDM had pronounced decreased selenium concentrations in erythrocytes as compared to controls. PMID- 10836531 TI - Does a relationship exist between ioduria, magnesiuria and calciuria? AB - We investigated the mutual relations between ioduria in the one hand, and calciuria, magnesiuria, and creatininuria, on the other hand in a randomly selected group of the population of the Czech Republic. The individual parameters were always determined in the sample of monitoring urine after night fasting, concentration according to the WHO/ICCIDD, we observed a parallel increase of calciuria, magnesiuria and creatininuria. The values of calciuria, magnesiuria and creatininuria correlated positively with ioduria both in children and in adults aged 6-93 years without any statistical effect of sex. PMID- 10836532 TI - Analysis of silicon in human tissues with special reference to silicone breast implants. AB - The increase, in the last two decades, in the application of silicones (polysiloxanes) and inorganic silicon compounds in medicine and the food industry, has exposed the human body to extensive contacts with these substances. Most silicone breast implants contain a gel consisting of a crosslinked silicone elastomer swollen by silicone oil (PDMS). Diffusion of PDMS through the silicone elastomer envelope and rupture of the envelope with release of the gel contents both occur clinically. The amount and distribution of silicone compounds in various tissues are key issues in the assessment of health problems connected with silicone implants. We have measured by GFAAS the Si content of tissues from normal and implant patients and the organic solvent extractable Si levels (assumed to be silicone), using careful control of sample collection and preparation. Whole blood levels were: implant patients mean 38.8 (SD 25.6) (microg/kg), controls mean 24.2 (SD 26.7) (microg/kg) in one study and subsequently 103.8 (SD 112.1) and 74.3 (SD 86.5) (microg/kg) in another study. Capsular tissue levels were: gel implants 25047 (SD 39313) (mg/kg of dry tissue), saline implants 20.0 (SD 27.3) (mg/kg of dry tissue) and controls 0.24 (SD 0.39) (mg/kg of dry tissue). Breast milk levels were: implant patients mean 58.7 (SD 33.8) (microg/kg), controls mean 51.1 (SD 31.0) (microg/kg); infant formula mean was 4.40 (mg/kg). Various precautions were undertaken to avoid Si contamination in this work, the most important being a) the use of a Class 100 laboratory for sample preparation and b) application of strict and elaborate washing procedure for specimen collection tools and laboratory plasticware. This data demonstrated that to properly interpret the importance of these numbers for human health, a larger study of "normal" levels of Si in human tissues should be undertaken and factors such as diet, water, race and geographical location should be considered. PMID- 10836533 TI - Preparation of selenium yeasts I. Preparation of selenium-enriched Saccharomyces cerevisiae. AB - Selenium (Se) is an essential micronutrient for human and animal organisms. Organic selenium complexes and selenium-containing amino acids are considered the most bioavailable. Under appropriate conditions yeasts are capable of accumulating large amounts of trace elements, such as selenium, and incorporating them into organic compounds. It has been found that introduction of water-soluble selenium salt as a component of the culture medium for yeasts produced by conventional batch processing results in a substantial amount of selenium being absorbed by the yeast. Using a culture medium supplemented with 30 microg/mL sodium-selenite added during the exponential growth phase results in selenium accumulation in the range of 1200-1400 microg/g dried baker's yeast (Saccharomyces cerevisiae) measured by ICP-AES method. In our previous studies it was shown that higher amounts of sodium-selenite in the culture medium have a strong inhibitory effect on the growth of this yeast. As a consequence of variations in cultivation conditions we obtained selenium yeast with different inorganic selenium content. The most important parameters influencing incorporated forms of selenium are pH value and dissolved oxygen level in the culture medium, and depending on these the selenium consumption rate of the yeast. A 0.40-0.50 mg/g h-1 specific selenium consumption rate was found to be appropriate to obtain selenium-enriched bakers' yeast of a high quality. Under suitable conditions the undesirable inorganic selenium content of the yeast could be suppressed to as low as 5-6% at the expense, however, of approximately a 20% decrease in the final biomass. PMID- 10836534 TI - Application of stripping voltammetry and microelectrodes in vitro biocompatibility and in vivo toxicity tests of AISI 316L corrosion products. AB - Adsorptive stripping voltammetric procedures, using mercury film microelectrodes, were optimised and applied to quantify total iron, chromium and nickel in samples of osteoblast-like cells culture medium and mice organs (liver, kidney and spleen) obtained from, respectively, in vitro and in vivo 316L stainless steel corrosion products biocompatibility and toxicity studies. The methods were based on the pre-concentration of the iron-catechol complex by adsorption at the potential of -1.80 V (vs. Ag/AgCl), of the chromium-diethylenetriaminepentaacetic acid complex at -1.00 V or -1.15 V (vs. Ag/AgCl) and of the nickel dimethylglyoxime complex at -0.70 V (vs. Ag/AgCl). The detection limits achieved for each metal ion (i) in the culture medium were 1.93x10(-8) mol/L Fe, 2.80x10( 10) mol/L Cr and 7.70x10(-9) mol/L Ni for a collection time of 30 s, 40 s and 10 s, respectively, and (ii) in the mice organ solutions were 1.37x10(-8) mol/L Fe, 1.54x10(-8) mol/L Cr and 1.58x10(-9) mol/L Ni for an adsorption time of 25 s, 25 s and 15 s, respectively. The accuracy of the proposed procedures was verified by comparison of the results obtained by adsorptive stripping voltammetry with those attained by atomic absorption spectrometry for the same set of samples and good agreement was found. The in vitro study showed that stainless steel corrosion products affect the expression of the osteogenic phenotype. The in vivo mice model, used to investigate the systemic effects provoked by the corrosion products per se, indicated that Fe, Cr and Ni are partially accumulated in the organs studied and that Ni induced the more significant morphological alterations. PMID- 10836535 TI - Protective effect of zinc on cadmium-induced micronuclei in V79 cells. AB - The induction of micronuclei (MN) in mitotically active cells has been widely used and promoted as a biological marker of exposure to environmental toxins. In our study the effect of zinc on cadmium genotoxicity was investigated in V 79 cells. The results indicate that cadmium chloride exposure for 24 h increased micronucleus frequency and the percentage of binucleated cells in dose-dependent manner. At the highest concentration of cadmium (50 microM Cd) 23 MN were found in 1000 cells. The protective effect of zinc on cadmium genotoxicity was investigated at lower concentrations (5-25 microM CdCl2). At 50 microM Cd, the number of MN increased significantly (16 MN). PMID- 10836536 TI - Photoimmunology, DNA repair and photocarcinogenesis. AB - In recent years major progress has been made in identifying the molecular mechanisms by which UV radiation modulates the immune system of the skin. From these studies it appears that the generation of DNA damage and the subsequent activation of DNA repair enzymes play a critical role in the generation of UV-B induced immunosuppression. These studies have made use of cells from both nucleotide excision repair (NER)-deficient individuals and mice. Results obtained from these studies have important clinical implications for DNA-repair-deficient patients in particular and for effective photoprotection of human skin in general. PMID- 10836537 TI - Key issues in the photochemistry and signalling-state formation of photosensor proteins. AB - Four families of photosensors (i.e., rhodopsins, phytochromes, xanthopsins and cryptochromes) exist, which vary widely in the degree to which we understand the molecular basis of their activity. Some of their members are ideal model systems for studying the structure-function relation of proteins, and the role of dynamics therein. The photochemistry of photosensor activation is based upon the cis <--> trans isomerization of the chromophore. This configurational transition leads to the formation of a signalling state of sufficient stability to communicate the presence of photons to a downstream signal-transduction partner. In the xanthopsins it has been demonstrated that the exact nature of this signalling state is strongly dependent on the mesoscopic context of the sensor protein. The cryptochromes appear to challenge the photoisomerization rule. PMID- 10836538 TI - Hypericin photosensitization of tumor and metastatic cell lines of human prostate. AB - We have investigated the photoactivating effect of hypericin on two cancer cell lines: PC-3, a prostatic adenocarcinoma non-responsive to androgen therapy and LNCaP, a lymphonodal metastasis of prostate carcinoma responsive to androgen therapy. The two cell lines are incubated for 24 h with hypericin at concentrations ranging from 0.001 to 0.3 microg/ml in cell culture medium. The cells are irradiated at 599 nm (fluence = 11 J/cm2) using a dye laser pumped by an argon laser. Hypericin exerts phototoxic effects on both cell lines, while it does not produce toxic effects in the absence of irradiation. These results suggest that photodynamic therapy (PDT) with hypericin could be an alternative approach to the treatment of prostatic tumors, and could be beneficial in tumors that are non-responsive to androgen therapy. PMID- 10836539 TI - Localisation and accumulation of a new carotenoporphyrin in two primary tumour models. AB - We have investigated the tumour-localising properties and in vivo fluorescence kinetics of a hexamethoxylated carotenqporphyrin (CP6) in two primary tumour models: UV-B-induced early skin cancer in hairless mice and chemically induced mucosal dysplasia in the rat palate. CP6 fluorescence kinetics are investigated by measuring in vivo fluorescence spectra and images of the mouse skin and the rat palate at different time points after injection. For the tumour-localising properties, microscopic phase-contrast and fluorescence images are recorded. The in vivo fluorescence kinetics in the mouse skin show localization of CP6 in the tumours. However, fluorescence microscopy images show that CP6 localises in the dermis and structures that are not related to the malignant transformation of the mouse skin. The fluorescence kinetics in the rat palate show a significant correlation between the degree of malignancy and the CP6 fluorescence build-up time in the palate. The microscopic images show that CP6 fluorescence localises in the connective tissue and not in the dysplastic epithelium. In conclusion, CP6 does not localise preferentially in (pre-) cancerous tissue in the two primary tumour models studied here, in contrast to reports about localisation of carotenoporphyrins in transplanted tumours. However, the CP6 build-up time in rat palates correlates with the degree of malignancy and this might possibly be a useful parameter in tumour detection. PMID- 10836540 TI - Diffuse component of solar ultraviolet radiation in tree shade. AB - The first set of quantitative data of diffuse erythemal UV and UV-A radiation in tree shade at a sub-tropical Southern Hemisphere latitude is presented. Over the summer, approximately 60% of the erythemal UV radiation in tree shade is due to the diffuse component. Similarly, approximately 56% of the UV-A radiation in tree shade is due to the diffuse component. In tree shade these diffuse UV percentages are relatively constant from the morning to noon to afternoon periods. In comparison, in full sun, there is a decrease in the percentage of diffuse UV from morning to noon to afternoon. The exposures to diffuse UV on a horizontal plane in tree shade between 9:00 EST and 15:00 EST are of the order of 4 MED (minimum erythemal dose) and 14 J cm(-2) for erythemal UV and UV-A, respectively. The high diffuse UV component in the shade may result in high UV exposures not only to unprotected parts of the body on a horizontal plane, but also in equally high UV irradiances to parts of the body, including the eyes and face, that are not UV protected. PMID- 10836541 TI - Oxidative stress and defence mechanisms of the freshwater cladoceran Daphnia longispina exposed to UV radiation. AB - The aim of the present study is to evaluate the occurrence of oxidative stress in the cladoceran Daphnia longispina exposed to UV-A and UV-B radiation. The activity of antioxidant enzymes and lipid peroxidation markers is investigated and the protective action of ascorbic acid determined. Results show differences in the lethality radioinduced by UV-A and UV-B. Both UV-A and UV-B exposure cause an important increase in malonaldehyde (MDA) concentration and catalase activity. Ascorbic acid addition reduces the MDA concentration, indicating that the oxidative stress caused by either UV-A or UV-B radiation can be controlled by antioxidants. The increase of the antioxidant enzymes may be a response mechanism to oxidative stress. PMID- 10836542 TI - An estimation of biological hazards due to solar radiation. AB - A spectrum evaluator based on four different dosimeter materials has been employed to estimate the spectral irradiances of solar radiation for exposed humans. The result is used to calculate the biologically effective irradiance using the erythemal action spectrum and a fish melanoma action spectrum. Measurements are made in winter at a sub-tropical site on the chest and shoulder of subjects during normal daily activities. Up to 95% of the total UV exposure received is in the UV-A waveband (320-400 nm). The UV-A waveband is found to contribute approximately 14% of the erythemal UV and 93% of the biologically effective UV for fish melanoma. Extrapolation to humans suggests that exposure to the UV-A band will contribute to photodamage in human skin during exposure to solar radiation. PMID- 10836543 TI - Immunochemical analysis of a photoreceptor protein using anti-IP3 receptor antibody in the unicellular organism, Blepharisma. AB - The blepharismin-200 kD protein complex of the ciliated protozoan Blepharisma is a novel type of photosensor responsible for the step-up photophobic response of the cell. In immunoblotting assays, the 200 kD protein is weakly cross-reacted with anti-inositol triphosphate receptor antibody (anti-IP3 R antibody). Indirect immunofluorescence assays show that the pigment granules in which the blepharismin-200 kD protein complex is localized are labelled by anti-IP3 R antibody. When the anti-IP3 R antibody or antisense oligonucleotide for IP3 receptor is introduced into the living cells of Blepharisma, both the photosensitivity of the cells and content of blepharismin-200 kD protein are reduced. The results suggest that the photoreceptor 200 kD protein is possibly an IP3 receptor-like protein. PMID- 10836544 TI - Preferential cytotoxicity for multidrug-resistant K562 cells using the combination of a photosensitizer and a cyanine dye. AB - The cyanine dye 1,1',3,3,3',3'-hexamethylindodicarbocyanine iodide (HIDC) protects K562 leukemia cells from photodynamic membrane damage caused by cis-di(4 sulfonatophenyl)diphenylporphine (TPPS2) and 420 nm light. This wavelength of light is chosen because it is absorbed by TPPS2, but not by HIDC. The photodynamic system studied may be useful as a model for antineoplastic therapy. A subline of K562 leukemia (K562/DOX), expressing the multidrug-resistance (MDR) phenotype, is found to accumulate smaller amounts of HIDC than the parent cell line and thus has less photoprotection. In the absence of added HIDC, the K562/DOX cell line is more resistant to photodynamic cytotoxicity than the K562 cell line. The resistance of the K562/DOX cell line is not due to a smaller accumulation of TPPS2 than the K562 cell line. However, when both cell lines are incubated with HIDC and TPPS2, and then exposed to light, the K562/DOX cell line becomes more sensitive to photodynamic cell damage than the K562 cell line. The combination of a photosensitizer with a cationic or lysomorphotropic photoprotector represents a novel strategy for the eradication of malignant cells expressing the MDR phenotype. PMID- 10836545 TI - Electrospray-mass spectrometry characterization and measurement of far-UV-induced thymine photoproducts. AB - Far-UV-induced formation of dimeric pyrimidine photoproducts within DNA is a major cause of the carcinogenic effects of solar light. The chemical structure of this class of lesion has been mostly determined by studies on model compounds. The present work is aimed at providing mass spectrometry data on the thymine thymine photoproducts, including the diastereoisomers of the cyclobutane dimer, the (6-4) adduct, the related Dewar valence isomer and the spore photoproduct. Fragmentation mass spectra of the modified bases, nucleosides, dinucleoside monophosphates and dinucleotides were recorded following electrospray ionization with either triple-quadrupolar or ion-trap detection. The results showed differences in fragmentation pattern between the different types of photoproducts. In addition, a drastic effect of the diastereoisometry was observed for the cyclobutane dimers. A sensitive detection technique has been developed for the analysis of dinucleoside monophosphate photoproducts by high performance liquid chromatography associated with mass spectrometry in the negative mode with multiple reaction-monitoring detection. PMID- 10836546 TI - Non-coherent visible and infrared radiation increase survival to UV (254 nm) in Escherichia coli K12. AB - Interactions between visible or infrared (IR) and ultraviolet (UV, 254 nm) radiation have been studied in E. coli. Pre-illumination with non-coherent monochromatic 446, 466, 570 and 685 nm radiation, as well as with polychromatic red and IR radiation at room temperature, leads to increased cell survival after a subsequent irradiation with UV light. In the thermic range of the spectrum (red and IR), IR but not red light pre-treatment is able to increase cell survival to a subsequent lethal heat (51 degrees C) challenge, suggesting that increased UV survival may be due to IR-induced heat-shock response. On the other hand, visible light-induced resistance may be due to a different mechanism, possibly involved with unknown bacterial light receptors. PMID- 10836547 TI - Photophysical properties of carborane-containing derivatives of 5,10,15,20 tetra(p-aminophenyl)porphyrin. AB - Absorption, fluorescence emission, fluorescence excitation spectra and fluorescence decay kinetics of carborane derivatives of 5,10,15,20-tetra(p aminophenyl)porphyrin have been investigated. Carborane derivatives are prepared by acylation of the amino groups of 5,10,15,20-tetra(p-aminophenyl)porphyrin by 9 carboranyl acetyl chloride. From the analysis of the absorption and fluorescence spectra, it is concluded that covalent linking of carborane molecules to the tetraphenylporphyrin molecule significantly changes the self-conjugated pi-system of the porphyrin macrocycle: positions of maxima of absorption and fluorescence spectra shift to the red region by 3-8 nm; the halfwidths of these bands are broadened by 2.5-5.0 nm; the relative intensity of the bands I-IV also changes. The fluorescence decay kinetics of the carborane derivatives are biexponential. According to the experimental data and model simulation, it is concluded that the intramolecular electron transfer proceeds from the porphyrin excited part of the molecule to carboranyls with a rate constant of 415 ps(-1) and efficiency of 0.16 0.8. Recombination of separated charges occurs within 1.4 ns. PMID- 10836548 TI - Effects of ultraviolet light on free and peptide-bound pyridinoline and deoxypyridinoline cross-links. Protective effect of acid pH against photolytic degradation. AB - Little is known about the photodegradation of pyridinoline (Pyd) and deoxypyridinoline (Dpd), which are two mature cross-links stabilizing collagen within extracellular matrix. In this study, highly purified free Pyd and Dpd cross-links have been degraded by irradiation with ultraviolet light and we have shown that photolysis varies with the pH value. Assessment of photolysis in basic (pH 9) and neutral (pH 7) solutions by high-performance liquid chromatography as well as by UV absorbance measurement indicates that both cross-links are degraded after a 24 h UV exposure, while in acidic solution (pH 3) only Dpd is photolysed, suggesting that acid pH provides major protection against Pyd photolysis. Photodegradation products have been studied by amino-acid and mass spectral analysis. Both methods confirm the lack of Pyd degradation in acid pH. Furthermore, amino-acid analysis allows us to identify hydroxylysine and lysine as a result of Pyd and Dpd photolysis, respectively, indicating that the mechanism of photodegradation involves the cleavage of the pyridinium ring on each side of the quaternary nitrogen. Finally, we have also studied the photolysis of different molecular species of type I collagen peptides, obtained by digestion with collagenase of demineralized turkey bone. Our results indicate that even when they are part of the structure of collagen peptide, Pyd and Dpd can be photolysed. However, we have shown that the larger the peptide is, the smaller are the effects of UV irradiation. PMID- 10836550 TI - Phototactic orientation mechanism in the ciliate Fabrea salina, as inferred from numerical simulations. AB - The marine ciliate Fabrea salina shows a clear positive phototaxis, but the mechanism by which a single cell is able to detect the direction of light and orient its swimming accordingly is still unknown. A simple model of phototaxis is that of a biased random walk, where the bias due to light can affect one or more of the parameters that characterize a random walk, i.e., the mean speed, the frequency distribution of the angles of directional changes and the frequency of directional changes. Since experimental evidence has shown no effect of light on the mean speed of Fabrea salina, we have excluded models depending on this parameter. We have, therefore, investigated the phototactic orientation of Fabrea salina by computer simulation of two simple models, the first where light affects the frequency distribution of the angles of directional changes (model M1) and the second where the light bias modifies the frequency of directional changes (model M2). Simulated M1 cells directly orient their swimming towards the direction of light, regardless of their current swimming orientation; simulated M2 cells, on the contrary, are unable to actively orient their motion, but remain locked along the light direction once they find it by chance. The simulations show that these two orientation models lead to different macroscopic behaviours of the simulated cell populations. By comparing the results of the simulations with the experimental ones, we have found that the phototactic behaviour of real cells is more similar to that of the M2 model. PMID- 10836549 TI - Chromosomes are target sites for photodynamic therapy as demonstrated by subcellular laser microirradiation. AB - The present investigation has been undertaken to examine the possibility that the cell nucleus, and specifically the genetic material, is a target site for photodynamic therapy. PTK2 and Hep-2 cells are pretreated with a medium containing 15 microg/ml (0.09 mM) 5-aminolevulinic acid (ALA). Individual fluorescence images are recorded for each selected cell using a cooled charge coupled device (CCD). A laser microbeam system generating 630 nm is used for subcellular-region irradiation of specific targets: chromosomes, the mitotic spindle, the perispindle region and the peripheral cytoplasm. Nuclei of interphase cells are also irradiated. Data comparing the sensitivities of the different subcellular microirradiation sites in ALA-treated mitotic cells demonstrate that under the irradiation conditions used, the chromosome is the most sensitive subcellular target followed by the perispindle region, the peripheral cytoplasm and spindle, and, lastly, the interphase nucleus. PMID- 10836551 TI - Photophysical studies of the pheophorbide a dimer. AB - This work reports on a monomer-dimer equilibrium state of pheophorbide a in solution. A methodology for controlling the equilibrium constant by use of temperature and solvent variation is described. The absorption spectrum of the dimer is calculated, using different prepared equilibria of monomer and dimer in solution. We propose that these aggregates provide a good model for understanding the dimerization process in tetrapyrroles. PMID- 10836552 TI - Child development and the PITS: simple questions, complex answers, and developmental theory. Person in the Street. AB - The enormous popular interest in the field of child development makes it incumbent upon developmental scientists to convey with care the complexity of development lest oversimplified popular accounts gain credibility. Recent attempted models of development do include the range of variables and complexities that need to be accommodated in accounting for development. A model is presented here that incorporates many of the elements of recent models but elaborates on the role of experience in relation to the constitutional, cultural, economic, and social factors that contribute to advantages and disadvantages in children's development. The importance of accommodating data from prior theoretical perspectives and the importance of the contributions from neuroimaging studies are discussed as they are critical for successful theory building in the field of child development. PMID- 10836553 TI - Toward a science for and of the people: promoting civil society through the application of developmental science. AB - Applied developmental science (ADS) is scholarship that seeks to advance the integration of developmental research with actions-policies and programs-that promote positive development and/or enhance the life chances of vulnerable children and families. Through this integration ADS may become a major means to foster a science for and of the people. It may serve as an exemplar of the means through which scholarship, with community collaboration, may contribute directly to social justice. In so doing, ADS helps shift the model of amelioration, prevention, or optimization research from one demonstrating efficacy to one promoting outreach. When this contribution occurs in the context of university community partnerships, ADS may serve also as a model of how higher education may engage policy makers, contribute to community capacity to sustain valued programs, and maintain and perpetuate civil society through knowledge-based, interinstitutional systems change. PMID- 10836554 TI - Does memory development belong on an endangered topic list? AB - Although memory has long been regarded as a central and well-defined topic within the field of cognitive development, developments in and related to the study of memory increasingly suggest that the study of memory needs to be situated in a number of broader conceptual and research contexts. Four of these contexts are identified here and the ways they accommodate memory phenomena are examined: (1) knowledge (what is remembered?); (2) comprehension (what does it mean?); (3) context/function (why remember?); and (4) strategy (how to remember?). Memories, it is suggested, are best examined as knowledge structures that are the product of efforts to understand and to know, and memorizing is a socially situated activity undertaken in the service of individual or social goals. PMID- 10836555 TI - The rebirth of children's learning. AB - Learning is a central part of children's lives, but the study of learning is a rather peripheral part of the field of cognitive development. Fortunately, this situation is starting to change; recent theoretical and methodological advances have sparked renewed interest in children's learning. This renewed interest has already yielded a set of consistent and interesting findings regarding how children learn, as well as intriguing proposals regarding the mechanisms that underlie the learning. Increasing our focus on children's learning promises to yield practical benefits as well as a more exciting field of cognitive development. PMID- 10836556 TI - The promise of dynamic systems approaches for an integrated account of human development. AB - After decades of theoretical fragmentation and insularity, a converging explanatory framework based on general scientific principles is an important goal for developmental psychology. Dynamic systems approaches may provide such a framework, using principles of self-organization to explain how novel forms emerge without predetermination and become increasingly complex with development. New trends in traditional theoretical families emphasize systemic, emergent processes, and these can now be explicated with principles of self-organization that apply to all natural systems. Self-organization thus provides a single explanation for the multiple facets of development, integrating diverse developmental viewpoints within a larger scientific perspective. PMID- 10836557 TI - Close interrelation of motor development and cognitive development and of the cerebellum and prefrontal cortex. AB - Motor development and cognitive development may be fundamentally interrelated. Contrary to popular notions that motor development begins and ends early, whereas cognitive development begins and ends later, both motor and cognitive development display equally protracted developmental timetables. When cognitive development is perturbed, as in a neurodevelopmental disorder, motor development is often adversely affected. While it has long been known that the striatum functions as part of a circuit with dorsolateral prefrontal cortex, it is suggested here that the same is true for the cerebellum and that the cerebellum may be important for cognitive as well as motor functions. Like prefrontal cortex, the cerebellum reaches maturity late. Many cognitive tasks that require prefrontal cortex also require the cerebellum. To make these points, evidence is summarized of the close co-activation of the neocerebellum and dorsolateral prefrontal cortex in functional neuroimaging, of similarities in the cognitive sequelae of damage to dorsolateral prefrontal cortex and the neocerebellum, of motor deficits in "cognitive" developmental disorders, and of abnormalities in the cerebellum and in prefrontal cortex in the same developmental disorders. PMID- 10836558 TI - Evolutionary developmental psychology. AB - Evolutionary developmental psychology is the study of the genetic and ecological mechanisms that govern the development of social and cognitive competencies common to all human beings and the epigenetic (gene-environment interactions) processes that adapt these competencies to local conditions. The basic assumptions and domains of this emerging field, as related to human life history and social and cognitive development, are outlined, as are implications for issues of importance in contemporary society. PMID- 10836559 TI - The role of context in the development of psychopathology: a conceptual framework and some speculative propositions. AB - Despite the explosion of studies assessing relations between various contextual factors and various forms of psychological disturbance, about the only firm conclusion one can draw regarding the environment's role in the development of psychopathology is that "bad" things have "bad" effects among some-but not all people, some-but not all-of the time. We argue that extant research has confused two different roles of context and suggest that (1) environmental factors act as nonspecific stressors in the elicitation of psychopathology by provoking autonomic arousal, with specificity of expressed psychopathology governed by individual differences in endogenous factors; and that (2) context is specific in affecting the course of psychopathology by influencing the extent to which the behavioral, affective, or cognitive components of the pathology are repeated. PMID- 10836560 TI - Functional brain development in infants: elements of an interactive specialization framework. AB - One future direction for cognitive development research involves a closer integration with our knowledge about the developing brain. I present a framework for analyzing and interpreting postnatal functional brain development in human infants. Three specific hypotheses contribute to this framework, within which a variety of phenomena associated with the neural basis of perception and cognition in normal and abnormal development can be characterized. PMID- 10836561 TI - On the importance of comparative research: the case of folkbiology. AB - If our goal as researchers is to understand the range and complexity of human conceptual development, increased attention to comparative research is essential. I draw on research in the domain of folkbiology-commonsense understandings of plants and animals--to argue that several lines of comparative research are needed to understand the acquisition of folkbiology in particular and conceptual development in general. First, comparisons are needed between children and adults within a given society. It is impossible to understand the process of conceptual development without a detailed look at adult endstates in a domain. Second, comparisons are needed between adult endstates in different contexts. For a complete understanding of conceptual development we must understand the range of variability of adult conceptual systems. Finally, comparisons are needed among children developing in different contexts. Such research complements comparative work on adults and would serve to distinguish between universal and particular patterns of development and thus to inform and constrain accounts of conceptual development. PMID- 10836562 TI - Beliefs about truth and beliefs about rightness. AB - Children's developing conceptions of what is right or proper are commonly studied without reference to concomitant changes in their understanding of beliefs, just as studies of young people's maturing grasp of the belief entitlement process ordinarily proceed separately from any examination of the value considerations that invest beliefs with meaning. In an effort to reverse these isolationist practices, a case is made for rereading the fact-value dichotomy that currently works to divide the contemporaneous literatures dealing with children's moral reasoning development and their evolving theories of mind. Findings from two research programs, in which children's beliefs about truth and rightness are combined, serve to illustrate the natural interdependence of these moral and epistemic matters. PMID- 10836563 TI - Grounding development in cognitive processes. AB - Developmentalists have made remarkable progress over the last several decades in detailing what children know at various points in development. Less progress has been made, however, in detailing the processes through which knowledge is realized in real-time tasks, or in detailing the processes of developmental change. We argue that the operating characteristics of perceiving and remembering provide a foundation for making progress on these issues in the next century. We include three examples applying these ideas to specific phenomena in early word learning. These examples illustrate how forming developmental hypotheses in terms of perceiving and remembering may bring new insights into specific phenomena as well as into how the ordinary operating characteristics of perceiving and remembering serve as bootstraps to more specialized and more abstract kinds of knowledge. PMID- 10836564 TI - An integrated model of emotion processes and cognition in social information processing. AB - Literature on the contributions of social cognitive and emotion processes to children's social competence is reviewed and interpreted in the context of an integrated model of emotion processes and cognition in social information processing. Neurophysiological and functional evidence for the centrality of emotion processes in personal-social decision making is reviewed. Crick and Dodge's model is presented as a cognitive model of social decision making, and a revised model is proposed into which emotion processes are integrated. Hypotheses derived from the proposed model are described. PMID- 10836565 TI - The future of infant categorization research: a process-oriented approach. AB - We call for a shift from a content-oriented approach to a process-oriented approach to the study of categorization in infancy. Although gains have been made in our understanding of infant categorization by evaluating the categories to which infants respond, further understanding of infant categorization, and how categorization changes with development, requires that we more directly assess infants' category formation. We argue that two directions for future research will continue to enhance our understanding of categorization in infancy (and beyond). First, contextual variations (e.g., the effects of task, stimuli, and other factors) on infants' categorization must be better understood. Second, we must more directly evaluate the kinds of information infants use when forming categories, as well as how their use of such information changes with age, task, and so forth. We argue that these two foci will provide clearer understanding of the origins and early development of categorization. PMID- 10836566 TI - Fatherhood in the twenty-first century. AB - The twentieth century has been characterized by four important social trends that have fundamentally changed the social cultural context in which children develop: women's increased labor force participation, increased absence of nonresidential fathers in the lives of their children, increased involvement of fathers in intact families, and increased cultural diversity in the U.S.. In this essay, we discuss how these trends are changing the nature of father involvement and family life, and in turn affecting children's and fathers' developmental trajectories. We end with an eye toward the twenty-first century by examining how the children of today will construct their expectations about the roles of fathers and mothers as they become the parents of tomorrow. This life-span approach to fatherhood considers the broader sociohistorical context in which fatherhood develops, and emphasizes the urgent need to consider mothers, fathers, and family structure in future research as we seek to understand and model the effects of parenting on children's development. PMID- 10836567 TI - A perception--action perspective on tool use development. AB - In this essay I argue for a new wave of research on tool use development. Advances in the literature on perception-action development hold important clues for how tool use unfolds in children. These advances suggest that tool use may be a more continuous developmental achievement than has been previously believed. On this view, tool use is rooted in the perception-action routines that infants employ to gain information about their environments. Although tools alter the properties of effector systems, children use tools to explore and change their environments, building on efforts that originate in infancy. Based on this approach, new research directions are suggested, including efforts designed to investigate the processes by which children detect and relate affordances between objects, coordinate spatial frames of reference, and incorporate early-appearing action patterns into instrumental behaviors. PMID- 10836568 TI - The legacy of early attachments. AB - The impact of early close relationships on psychological development is one of the enduring questions of developmental psychology that is addressed by attachment theory and research. This essay evaluates what has been learned, and offers ideas for future research, by examining the origins of continuity and change in the security of attachment early in life, and its prediction of later behavior. The discussion evaluates research on the impact of changing family circumstances and quality of care on changes in attachment security, and offers new hypotheses for future study. Considering the representations (or internal working models) associated with attachment security as developing representations, the discussion proposes that (1) attachment security may be developmentally most influential when the working models with which it is associated have sufficiently matured to influence other emerging features of psychosocial functioning; (2) changes in attachment security are more likely during periods of representational advance; and (3) parent-child discourse and other relational influences shape these developing representations after infancy. Finally, other features of early parent-child relationships that develop concurrently with attachment security, including negotiating conflict and establishing cooperation, also must be considered in understanding the legacy of early attachments. PMID- 10836569 TI - The motor core of speech: a comparison of serial organization patterns in infants and languages. AB - Comparison of serial organization of infant babbling and early speech with that of 10 languages reveals four movement-related design features reflecting a deep evolutionary heritage: (1) the cyclical consonant-vowel alternation underlying the syllable, a "Frame" for speech consisting of mandibular oscillation, possibly evolving from ingestive cyclicities (e.g., chewing) via visuofacial communicative cyclicities (e.g., lipsmacks); (2) three intracyclical consonant-vowel co occurrence preferences reflecting basic biomechanical constraints-coronal consonants-front vowels, dorsal consonants-back vowels, and labial consonants central vowels; (3) a developmental progression from above-chance to below-chance levels of intercyclical consonant repetition; (4) an ease-related labial consonant-vowel-coronal consonant sequence preference for word initiation. These design features presumably result from self-organizational responses to selection pressures, primarily determined by motor factors. No explanation for these design features is available from Universal Grammar, and, except for feature 3, perceptual-motor learning seems to have only a limited causal role in acquisition of any design feature. PMID- 10836570 TI - A secure base from which to explore close relationships. AB - The theory of attachment as a secure-base relationship integrates insights about affect, cognition, and behavior in close relationships across age and culture. Empirical successes based on this theory include important discoveries about the nature of infant-caregiver and adult-adult close relationships, the importance of early experience, and about stability and change in individual differences. The task now is to preserve these insights and successes and build on them. To accomplish this, we need to continually examine the logic and coherence of attachment theory and redress errors of emphasis and analysis. Views on attachment development, attachment representation, and attachment in family and cross-cultural perspective need to be updated in light of empirical research and advances in developmental theory, behavioral biology, and cognitive psychology. We also need to challenge the theory by formulating and testing hypotheses which, if not confirmed, would require significant changes to the theory. If we can accomplish these tasks, prospects for important developments in attachment theory and research are greater than ever, as are the prospects for integration with other disciplines. PMID- 10836571 TI - Effect size, practical importance, and social policy for children. AB - Real decisions for real children are influenced by the papers developmentalists write, regardless of whether we ever intended our papers to be used in the policy arena. Yet most social scientists seldom analyze data in ways that are most useful to policymakers. The primary purpose of this paper is to share three ideas concerning how to evaluate the practical importance of a finding or set of findings. First, for research to be most useful not only in the policy arena but also more generally, significance tests need to be accompanied by effect size estimates. The practical importance of an effect size depends on the scientific context (i.e., measurement, design, and method) as well as the empirical literature context. Second, researchers need to use all existing data when weighing in on a policy debate; here, meta-analyses are particularly useful. Finally, researchers need to be careful about embracing null or small findings, because effects may well be small due to measurement problems alone, particularly early in the history of a research domain. PMID- 10836572 TI - Science, policy, and practice: three cultures in search of a shared mission. AB - Research on child development, the design of social policies, and the delivery of human services for children and families reflect three related yet separate cultures. The capacity to navigate across their borders, to understand their different rules of evidence, to speak their distinctive languages, and to achieve credibility in all three worlds while maintaining a sense of intellectual integrity in each, requires respect for their differences and a commitment to their shared mission. The transmission of knowledge from the academy to the domains of social policy and practice is a formidable task. This challenge could be facilitated by a simple taxonomy that differentiates established knowledge from both reasonable hypotheses and unwarranted or irresponsible assertions that are made in the name of science. An investment in effective "cross-cultural" translation offers a potent strategy for enhancing both the generation of new research and the application of cutting-edge knowledge to make a difference in the lives of children and their families. PMID- 10836573 TI - Family poverty, welfare reform, and child development. AB - Our review of research suggests that family poverty has selective effects on child development. Most important for policy are indications that deep or persistent poverty early in childhood affects adversely the ability and achievement of children. Although the 1996 welfare reforms have spurred many welfare-to-work transitions, their time limits and, especially, sanctions are likely to deepen poverty among some families. We suggest ways policies might be aimed at preventing either economic deprivation itself or its effects. PMID- 10836574 TI - The future of applied child development research and public policy. AB - After reviewing a brief general history of applied child development research, this paper suggests that in the future we should study questions that society needs to answer as well as questions that might contribute to theory, and that our research methods need to be adjusted to match these types of questions. Further, academics are urged to broaden their audience from a nearly exclusive focus on other academics to a focus on the three ps--practitioners, policymakers, and the public--and to recognize that scholarship is packaged differently for these audiences. Finally, it is suggested that applied child development research should market as well as sell, partner with nonacademic groups, disseminate results more vigorously, and focus efforts on local as well as national issues. PMID- 10836575 TI - New directions in analyses of parenting contributions to children's acquisition of values. AB - Traditional theories of how children acquire values or standards of behavior have emphasized the importance of specific parenting techniques or styles and have acknowledged the importance of a responsive parent-child relationship, but they have failed to differentiate among forms of responsiveness, have stressed internalization of values as the desired outcome, and have limited their scope to a small set of parenting strategies or methods. This paper outlines new directions for research. It acknowledges the central importance of parents and argues for research that (1) demonstrates that parental understanding of a particular child's characteristics and situation rather than use of specific strategies or styles is the mark of effective parenting; (2) traces the differential impact of varieties of parent responsiveness; (3) assesses the conditions surrounding the fact that parents have goals other than internalization when socializing their children, and evaluates the impact of that fact; and (4) considers a wider range of parenting strategies. PMID- 10836576 TI - Criteria for evaluating the significance of developmental research in the twenty first century: force and counterforce. AB - Since its birth approximately 100 years ago, the field of child development has undergone fluctuations in the criteria used to determine which research topics are more or less worthy of study. The purpose of this paper is to identify the forces that influence how developmental research is prioritized and evaluated and how these influences are changing as we enter the new millennium. We do so by considering the developmental researcher in context and suggest that there will be increasing pressure to use new criteria when assessing the significance of twenty-first-century developmental science. We review the three most commonly used forms of research validity--internal, external, and ecological--and then identify new research validities that we believe are likely to play increasingly important roles in the next millennium. We also argue that many developmental scientists will increasingly be pressured by forces that are external to the traditional research environment and that these forces will shape the ways in which the significance of developmental research is evaluated. PMID- 10836577 TI - Integrating basic and applied developmental research: a new model for the twenty first century. AB - Until recently, basic and applied research agendas in the field of child development have followed separate paths. One reason the two have not merged is that the objectives of basic and applied research are often seen as incompatible. In this paper, we argue that researchers can simultaneously achieve the objectives of advancing basic knowledge and addressing applied problems within a single research program. We provide a framework for this perspective by first looking back at historical trends of basic and applied developmental research and then looking forward at potential new approaches for integrating basic and applied research. We use our own research on perception of affordances and unintentional childhood injuries to illustrate how researchers might implement these strategies for integrating basic and applied research. We conclude by discussing how we might extend this integration further to include nontraditional classes of application. PMID- 10836578 TI - Beyond words: the importance of gesture to researchers and learners. AB - Gesture has privileged access to information that children know but do not say. As such, it can serve as an additional window to the mind of the developing child, one that researchers are only beginning to acknowledge. Gesture might, however, do more than merely reflect understanding-it may be involved in the process of cognitive change itself. This question will guide research on gesture as we enter the new millennium. Gesture might contribute to change through two mechanisms which are not mutually exclusive: (1) indirectly, by communicating unspoken aspects of the learner's cognitive state to potential agents of change (parents, teachers, siblings, friends); and (2) directly, by offering the learner a simpler way to express and explore ideas that may be difficult to think through in a verbal format, thus easing the learner's cognitive burden. As a result, the next decade may well offer evidence of gesture's dual potential as an illuminating tool for researchers and as a facilitator of cognitive growth for learners themselves. PMID- 10836579 TI - The ontogeny of phonological categories and the primacy of lexical learning in linguistic development. AB - In this paper, we draw on recent developments in several areas of cognitive science that suggest that the lexicon is at the core of grammatical generalizations at several different levels of representation. Evidence comes from many sources, including recent studies on language processing in adults and on language acquisition in children. Phonological behavior is influenced very early by pattern frequency in the lexicon of the ambient language, and we propose that phonological acquisition might provide the initial bootstrapping into grammatical generalization in general. The phonological categories over which pattern frequencies are calculated, however, are neither transparently available in the audiovisual signal nor deterministically fixed by the physiological and perceptual capacities of the species. Therefore, we need several age-appropriate models of how the lexicon can influence a child's interactions with the ambient language over the course of phonological acquisition. PMID- 10836580 TI - Social life in cultures: judgements, conflict, and subversion. AB - We present a developmentally grounded perspective on the study of social life in cultures that examines the sources of the coexistence of sociability and opposition, harmony and conflict, acceptance and critique. The coexistence of these elements, it is proposed, is systematically connected both to the development of distinct domains of thought and to the dynamics of participation in more and less hierarchical social relationships in society. PMID- 10836581 TI - Depending on the kindness of strangers: current national data initiatives and developmental research. AB - This article provides a brief review of current large-scale, longitudinal data collection initiatives focusing on children. These studies will be available for secondary data analyses in the twenty-first century. In addition to child outcome data, process-oriented information is being collected on child-parent interactions, quality of child care, elementary school teacher reports and classroom observations, accessibility and use of health, educational and social services, parental mental health, family violence, fathering, parental residence patterns, income and income sources, child support, employment patterns, and community characteristics. Several of these studies are randomized trials of the efficacy of early childhood intervention services and housing mobility programs. The usefulness of these efforts for exploring policy-relevant issues (child support enforcement, work requirements for welfare recipients, antipoverty strategies, housing subsidies and relocation, availability of child care, child care subsidies) are discussed. PMID- 10836582 TI - A model of binocular stereopsis including a global consistency constraint. AB - The binocular correspondence problem was solved by implementing the uniqueness constraint and the continuity constraint, as proposed by Marr and Poggio [Marr D, PoggioT (1976) Science 194: 283-287]. However, these constraints are not sufficient to define the proper correspondence uniquely. With these constraints, random-dot stereograms (RDSs), consisting of the periodic textures in each image, are treated as a correspondence of surfaces composed of patches of alternating values of disparity. This is quite different from the surface we perceive through the RDSs, that is a surface characterized by a single depth. Because these constraints are local, they cannot produce the global optimum of correspondence. To obtain the global optimum of correspondence, we propose a model of binocular stereopsis in which a global measure of correspondence is explicitly employed. The model consists of two hierarchical systems. First, the lower system processes various correspondences based on the uniqueness constraint. Second, the higher system provides a global measure of correspondence for the disparity in question. The higher system uniquely determines the global optimum of correspondence in the lower system through the recurrent loop between hierarchical systems. The convergence of the recurrent loop is determined by the consistency between the hierarchical systems. The condition is termed the 'global consistency constraint. PMID- 10836583 TI - Nonlinear time-course of lumbar muscle fatigue using recurrence quantifications. AB - Isometric skeletal muscle fatigue is usually assumed to be a linear process based upon the monotonic decrease in spectral frequency of the EMG. Since spectral analysis by fast Fourier transform (FFT) constitutes a linear transformation of the data, the present study was designed to reevaluate the time-course of muscle fatigue with a nonlinear tool, recurrence quantification analysis (RQA). Surface EMG recordings were obtained from the multifidus muscle of 17 human subjects during isometric posture-holding of the upper torso. The process of muscle fatigue was found to be linear for 59% of the subjects by FFT criteria, but nonlinear for 76% by RQA criteria. As a demonstrative control, both slow and fast transients occurring within a nonlinear mathematical process could be accurately depicted by RQA, but not by FFT. It is concluded that assessment of EMG patterns by nonlinear techniques can give insight into the time-course of fatiguing muscles attributed to the summation of several nonlinear and competing processes. PMID- 10836584 TI - Recursive implementations of temporal filters for image motion computation. AB - Efficient algorithms for image motion computation are important for computer vision applications and the modelling of biological vision systems. Intensity based image motion computation proceeds in two stages: the convolution of linear spatiotemporal filter kernels with the image sequence, followed by the non-linear combination of the filter outputs. If the spatiotemporal extent of the filter kernels is large, then the convolution stage can be very intensive computationally. One effective means of reducing the storage required and computation involved in implementing the temporal convolutions is the introduction of recursive filtering. Non-recursive methods require the number of frames of the image sequence stored at any given time to be equal to the temporal extent of the slowest temporal filter. In contrast, recursive methods encode recent stimulus history implicitly in the values of a small number of variables updated through a series of feedback equations. Recursive filtering reduces the number of values stored in memory during convolution and the number of mathematical operations involved in computing the filters' outputs. This paper extends previous recursive implementations of gradient- and correlation-based motion analysis algorithms [Fleet DJ, Langley K (1995) IEEE PAMI 17: 61-67; Clifford CWG, Ibbotson MR, Langley K (1997) Vis Neurosci 14: 741-749], describing a recursive implementation of causal band-pass temporal filters suitable for use in energy- and phase-based algorithms for image motion computation. It is shown that the filters' temporal frequency tuning curves fit psychophysical estimates of the temporal properties of human visual filters. PMID- 10836585 TI - Modelling of congenital nystagmus waveforms produced by saccadic system abnormalities. AB - Models of the mechanisms of normal eye movements are typically described in terms of the block diagrams which are used in control theory. An alternative approach to understanding the mechanisms of normal eye movements involves describing the eye movement behaviour in terms of smooth changes in state variables. The latter approach captures the burst cell firing against motor error (difference between target gaze angle and current gaze angle) phase plane behaviour which is found experimentally and facilitates the modelling of variations in burst cell behaviour. A novel explanation of several types of congenital nystagmus waveforms is given in terms of a saccadic termination abnormality. PMID- 10836586 TI - Visual channels, Hebbian assemblies and the effect of Hebb's rule. AB - A model of sensory learning is proposed that is based upon Hebb's rule, where Hebb's rule has been generalised by introducing a stabilising function representing some feedback process within or at the adapting (cortical) neuron, preventing synaptic weights from increasing without limit. It will be shown that neurons adapting according to this stabilised Hebb rule will turn into a matched filter for that part of the stimulus pattern that covers the receptive field of a neuron. It follows that the presentation of a stimulus pattern may imply the formation of a set of neurons with overlapping receptive fields, where each neuron has adapted to a certain part of the stimulus. Making simplifying assumptions about the detection process, the model will be illustrated, fitting it to data from Meinhardt and Mortensen [Meinhardt G, Mortensen U (1998) Biol Cybern 79:413425] which are not compatible with the classical matched filter model introduced by Hauske et al. PMID- 10836587 TI - Analysis of recurrent cortico-basal ganglia-thalamic loops for working memory. AB - We analyse a simplified form of the frontal lobe architecture of cortico-basal ganglia-thalamo-cortical loops to determine the manner in which they can learn temporal sequences as part of working memory activity. In particular, we consider how the temporal duration of activity can arise in this setting. We start from a hard-wired version in which temporally extended activity is created by the 'long' loop of cortex --> basal ganglia --> thalamus --> cortex, and show it arises from a near saddle-node bifurcation. The manner in which the transition between patterns occurs is also considered. This is then extended to analyse the temporal sequence storage and regeneration abilities of trained networks with a similar architecture. The temporal dynamics of this activity is also analysed. Implications of this for other working memory activities and for understanding the architecture of the frontal lobes are discussed in conclusion. PMID- 10836588 TI - Assessment of the adiabatic transformability hypothesis in a ball-bouncing task. mbrodel@freewwweb.com. AB - The adiabatic transformability hypothesis for non-conservative, non-rate-limited biological systems put forward by Kugler and Turvey [Kugler PN, Turvey MT (1987) Information, natural law, and the self-assembly of rhythmic movements, Erlbaum, Hillsdale, NJ] is evaluated in a ball-bouncing task as a function of skill level. We hypothesized that, when a basketball player increases or decreases the frequency of his limb motion in a vertical ball-bouncing task, the transformation should be characterized as adiabatic. Confirmation of this hypothesis would lend support to the body of knowledge that suggests that physical-biological laws guide the behavior of people engaged in motor tasks, including their acquisition of skill. We videotaped and analyzed four participants - two intermediates and two experts - bouncing a ball from various heights, and measured the energy and kinematic relations of the ball and the participants' body segments. The task presents a challenge to certain predictions of the adiabatic hypothesis in evaluating changes in movement velocity (v) or frequency (f), and energy as adiabatic transformations. Among these are the constant relation between kinetic energy (Ek) per cycle and v, constancy of energy dissipated per cycle (Et) over changes in v and Ek, the ratio of Ek to Et per cycle (the "Q" values), and the relation of amplitude to v. From our observations, which are examined in regard to the insights of Kugler and Turvey about the relation of Ehrenfest's adiabatic theorem to biological systems, we confirm the basic adiabatic character of the task in analyzing both the ball alone and the relations of various body segments. In the segmental frame of reference, we found evidence of differences in energy kinematic relations in the Q values between skill levels. PMID- 10836589 TI - Accuracy of MR imaging for detecting epiphyseal extension of osteosarcoma. AB - BACKGROUND: Too few patients are receiving epiphyseal-sparing limb salvage procedures for osteosarcoma. OBJECTIVE: To determine how magnetic resonance (MR) imaging can best predict the epiphyseal extension of osteosarcoma. MATERIALS AND METHODS: Forty children underwent complete pretreatment static and dynamic contrast-enhanced MR imaging (DEMRI). Static MR images [T1-weighted and short tau inversion recovery (STIR)] of the epiphyses were read in three ways: (1) for suspicion of any abnormality (tumor or edema), (2) for suspicion of tumor, excluding suspected edema, and (3) validating the second method by using a scale to rate the likelihood of tumor. Presentation imaging was compared to histopathologic findings after chemotherapy and resection. The receiver operating characteristic (ROC) method was used to analyze the scaled ratings of static MR and DEMRI values. RESULTS: At delayed resection, 20 of 40 children with osteosarcoma had confirmed epiphyseal tumor; however, 32 epiphyses were abnormal on STIR and 28 abnormal on T1. Differentiating suspected tumor from edema increased the accuracy to an Az (area under the ROC curve) of 0.94 for both T1 weighted and STIR static sequences. T1-weighted MR had better specificity and STIR better sensitivity at any given rating. DEMRI was slightly less accurate (Az = 0.90). CONCLUSION: Static MR imaging most accurately detected epiphyseal extension of osteosarcoma when readers distinguished suspected tumor from edematous or normal tissue. PMID- 10836590 TI - Computers in radiology. The sedation, analgesia, and contrast media computerized simulator: a new approach to train and evaluate radiologists' responses to critical incidents. AB - BACKGROUND: Awareness and preparedness to handle sedation, analgesia, and contrast-media complications are key in the daily radiology practice. OBJECTIVE: The purpose is to create a computerized simulator (PC-Windows-based) that uses a graphical interface to reproduce critical incidents in pediatric and adult patients undergoing a wide spectrum of radiologic sedation, analgesia and contrast media complications. MATERIALS AND METHODS: The computerized simulator has a comprehensive set of physiologic and pharmacologic models that predict patient response to management of sedation, analgesia, and contrast-media complications. Photorealistic images, real-time monitors, and mouse-driven information demonstrate in a virtual-reality fashion the behavior of the patient in crisis. RESULTS: Thirteen pediatric and adult radiology scenarios are illustrated encompassing areas such as pediatric radiology, neuroradiology, interventional radiology, and body imaging. The multiple case scenarios evaluate randomly the diagnostic and management performance of the radiologist in critical incidents such as oversedation, anaphylaxis, aspiration, airway obstruction, apnea, agitation, bronchospasm, hypotension, hypertension, cardiac arrest, bradycardia, tachycardia, and myocardial ischemia. The user must control the airway, breathing and circulation, and administer medications in a timely manner to save the simulated patient. On-line help is available in the program to suggest diagnostic and treatment steps to save the patient, and provide information about the medications. A printout of the case management can be obtained for evaluation or educational purposes. CONCLUSION: The interactive computerized simulator is a new approach to train and evaluate radiologists' responses to critical incidents encountered during radiologic sedation, analgesia, and contrast-media administration. PMID- 10836591 TI - The use of FDG-PET and CT for the staging of adrenocortical carcinoma in children. PMID- 10836592 TI - Radiology reporting in an academic children's hospital: what referring physicians think. AB - Transcribed reports are the radiologist's most conspicuous and enduring product, yet relatively little investigation of report quality has been undertaken by radiologists. This paper reports the results of a survey of 266 referring physicians in an academic children's hospital regarding their needs and assessments of the quality of radiology reporting. The results outline the range and relative importance of report features from referring physicians' points of view, provide specific suggestions for how to improve reporting performance, and generally indicate that reporting should receive more attention in training and practice than it currently does. PMID- 10836593 TI - Fibrocartilaginous mesenchymoma of bone: case report and review of the literature. AB - Fibrocartilaginous mesenchymoma of bone is a rare primary neoplasm. Our literature search produced only 12 previously reported cases. Radiographic and computed tomography (CT) findings have been described, but the magnetic resonance imaging (MRI) appearance has not been reported previously. We report a patient with fibrocartilaginous mesenchymoma of the ilium and describe the imaging findings on conventional radiography, bone scan, CT, and MRI. PMID- 10836594 TI - Needle localization of small pediatric tumors for surgical biopsy. AB - BACKGROUND: Small pediatric tumors may be difficult to biopsy or resect. OBJECTIVE: To examine the benefits of needle localization of a variety of small pediatric tumors before surgical biopsy or excision. MATERIALS AND METHODS: Seven patients aged 1-19 years underwent 12 procedures for needle localization of suspected tumor. Two patients had undergone previous biopsies without needle localization with negative results. Computed tomography (four patients) or ultrasonography (three patients) guided needle placement. Each patient had suspected tumor(s) in 1-3 anatomical sites, including thigh (7), lung (2), parasacral region (2), and iliac bone (1). RESULTS: All 12 lesions (9 less than 1 cm3 in volume) were successfully localized for excision or biopsy. Three small (<1 cm3) soft-tissue lesions (two ganglioneuroblastomas and one ganglioneuroma) were excised from one patient, a 0.65-cm3 residual soft-tissue sarcoma from another; and recurrent bilateral teratomas from a third. Two peripheral primitive neuroectodermal tumors were excised with positive margins from a fourth patient. Two lesions contained only fibrosis, as determined by histopathology. Two other patients underwent thoracoscopic removal of lung metastases that were less than 0.1 cm3. CONCLUSION: Needle localization allows effective and less invasive excision or biopsy of a variety of small pediatric soft-tissue lesions. PMID- 10836595 TI - Cranial MRI in neonatal hypernatraemic dehydration. AB - Severe neonatal hypernatraemia is a life-threatening electrolyte disorder because of its neurological complications. These are brain oedema, intracranial haemorrhages, haemorrhagic infarcts and thromboses. There are few reports concerning the radiological findings in the central nervous system in severe neonatal hypernatraemia. Cranial MRI findings in hypernatraemia have been reported in an older child, but have not been described in newborn infants. We report the cranial MRI findings in a newborn infant with acute renal failure and severe hypernatraemia. PMID- 10836596 TI - Langerhans' cell histiocytosis presenting with an intracranial epidural hematoma. AB - An 8-year-old boy developed vomiting and severe headache following minor head trauma. A CT scan of the head demonstrated a lytic lesion of the skull and adjacent epidural hematoma. Surgical evacuation and removal of the skull lesion and hematoma were carried out, and pathologic evaluation resulted in a diagnosis of Langerhans' cell histiocytosis (LCH). Epidural involvement of Langerhans' cell histiocytosis is very rare, and we report the first case of LCH presenting as an intracranial epidural hematoma. PMID- 10836597 TI - Role of MR imaging in the diagnosis of complicated arachnoid cyst. AB - BACKGROUND: Arachnoid cysts are intra-arachnoidal cerebrospinal fluid collections that are usually asymptomatic, however, they may become acutely symptomatic due to enlargement of the cyst or the presence of hemorrhage. MATERIALS AND METHODS: We report a case of a child presenting with a 3-week history of headaches, nausea, and vision problems. There was no history of trauma. MRI clearly demonstrated a left middle cranial fossa arachnoid cyst with associated subacute intracystic and subdural hematoma that was causing mass effect and required surgery. RESULTS: This lesion was isodense to the brain on CT. CONCLUSION: We focus on the importance of MR imaging in the differentiation of these subacute/early chronic hemorrhagic collections that may be overlooked with CT. PMID- 10836598 TI - The use of oral pentobarbital sodium (Nembutal) versus oral chloral hydrate in infants undergoing CT and MR imaging--a pilot study. AB - BACKGROUND: Chloral hydrate, a commonly used oral sedative for infants undergoing imaging examinations, has a bitter taste and requires relatively large volume, provoking unpleasant reactions from the infants. Experience with an alternative sedative, oral pentobarbital (Nembutal), has not been reported for infants. OBJECTIVE: To compare patient acceptance of oral Nembutal and oral chloral hydrate for sedation of infants up to 12 months of age. METHODS AND MATERIALS: Fifty-four infants (mean age: 7 months) were prospectively enrolled. Parents chose Nembutal, chloral hydrate, or no preference. Thirty-eight infants received Nembutal (4-6 mg/kg) mixed with cherry syrup and 16 received chloral hydrate (50 100 mg/ kg). We recorded infant's acceptance of sedative, parental impression of infant's acceptance, time to sedation, time to discharge, adverse effects, parental preference of future sedative. RESULTS: Infant acceptance and parental impression were better for Nembutal (P < 0.0001). Fewer parents in the Nembutal group preferred another sedative (P = 0.05). There was a trend toward shorter time to discharge with Nembutal (P = 0.03). There were no adverse effects in either group. One infant failed to sedate with Nembutal. CONCLUSIONS: Compared with chloral hydrate, oral Nembutal has significantly better acceptance by infants and parents, equal effectiveness, and may result in a shorter time to discharge. PMID- 10836599 TI - Congenital intrahepatic portohepatic venous shunt: treatment with coil embolisation. AB - Congenital abnormalities of the portal venous system are rare. There are few radiological descriptions of intrahepatic portosystemic venous shunt detected in the perinatal period. We report a congenital portosystemic shunt that was detected by US and treated with coil embolisation in the neonatal period. PMID- 10836600 TI - Sonographic guidance of air enema for intussusception reduction in children. AB - BACKGROUND: Fluoroscopically guided air reduction of intussusception is a well accepted technique. There are only two previous reports in which US has been used to monitor pneumatic reduction. OBJECTIVE: To assess the ability of US to monitor the success of air reduction of intussusception. MATERIALS AND METHODS: Sonographically guided air-enema reduction of intussusception in 199 children. In phase I (11 children), the success or failure of reduction was confirmed by fluoroscopy. In phase II (188 children), complete reduction was confirmed by clinical improvement of the child and repeat sonography 1 h later showing no persistent intussusception. RESULTS: In phase I, fluoroscopy confirmed the accuracy of US in all 11 children. In phase II, the success rate of initial reduction was 95%. Following successful reduction, US repeated 1 h later showed no recurrence of intussusception in 92%. In ten (5%) of 188, initial reduction was unsuccessful; fluoroscopically guided air reduction successfully reduced only three of these ten failures. CONCLUSIONS: Air enema guided by US is a practical and reliable technique for the reduction of intussusception. PMID- 10836601 TI - Progressive liver calcifications in neonatal coxsackievirus infection. AB - Coxsackievirus group B can cause a severe systemic disease in the perinatal period. Severe manifestations like meningitis, encephalitis, hepatitis, and myocarditis have been previously reported. A case of a twin neonate infected by coxsackievirus group B is described, who developed progressive extensive hepatic calcifications demonstrated by ultrasound and computed tomography with follow-up. Hepatic calcifications in coxsackievirus infection have not been previously reported. PMID- 10836602 TI - Multiple unilateral renal cysts in two children. AB - We report two children who presented with multiple renal cysts involving only one kidney and in whom there was no family history of renal disease and who did not have syndromes known to be associated with renal cystic disease. This unilateral involvement may represent a distinct entity, which has only been previously described in three cases; however, long-term follow-up will be needed to confirm this hypothesis. We illustrate the sonographic and computed tomographic findings, and the differential diagnosis is discussed. PMID- 10836603 TI - Sialoblastoma and hepatoblastoma in a neonate. AB - We report a case of salivary gland neoplasm and associated hepatoblastoma. The sialoblastoma was diagnosed by prenatal sonography; however, the hepatoblastoma was imaged post-operatively. Prior knowledge could have prevented a subsequent operation, including the additional risks of repeat anesthesia. We suggest that if a sialoblastoma is in the differential diagnosis, then additional imaging may be indicated because of the possibility of associated lesions. PMID- 10836604 TI - Myocardial perfusion in children with sickle cell anaemia. AB - BACKGROUND: Myocardial ischaemia is an unexpected complication with potentially serious clinical damages in patients with sickle cell anaemia (SCA). Conventional techniques, such as exercise testing and echocardiography, have low sensitivity and specificity for the detection of myocardial ischaemia in patients with SCA. OBJECTIVE: To assess myocardial perfusion using thallium-201 (201Tl) single photon emission computed tomography (SPECT) in children with SCA. MATERIALS AND METHODS: Eight patients (11.5 +/- 5.0 years, mean +/- SD) who were free of cardiac symptoms were studied. Myocardial perfusion was assessed by 201Tl-SPECT after stress and 3 h later after a further injection. Left ventricular ejection fraction (LVEF) was assessed by equilibrium radionuclide angiography. RESULTS: Myocardial perfusion was abnormal in three of eight patients: two had reversible defects and one had a fixed defect. The mean LVEF was 53 +/- 8%. There was no relationship between myocardial perfusion and LVEF. CONCLUSIONS: Treatment of asymptomatic myocardial ischaemia remains unclear, and more aggressive therapy of the haematological disease should be considered. PMID- 10836606 TI - Thyroid follicular carcinoma: a case report. AB - We present an 8-year-old boy with thyroid follicular carcinoma. Differentiated thyroid follicular carcinoma is very uncommon in childhood and is not usually suspected as the explanation of a thyroid nodule during infancy. The purpose of our paper is to demonstrate the imaging findings in a child and to emphasise the importance of including this tumour in the differential diagnosis of thyroid nodules in children. PMID- 10836605 TI - Brucellosis: unusual presentations in two adolescent boys. AB - Two boys presented with variable signs and symptoms of infectious disease that challenged diagnosis. One of the two patients had aortic valve vegetations and lower extremity aneurysms, and the other had calvarial osteomyelitis, epidural abscess, pleural effusions, and pulmonary nodules. Only after a battery of bacterial and fungal agglutination tests was the unsuspected diagnosis made in each of brucellosis from Brucella canis. PMID- 10836607 TI - MR imaging of juvenile granulosa cell tumour of the ovary: a case report. PMID- 10836608 TI - A de novo glycine substitution mutation in the collagenous domain of COL7A1 in dominant dystrophic epidermolysis bullosa. AB - Dystrophic epidermolysis bullosa (DEB) is a hereditary mechanobullous disorder characterized by fragility of the skin and mucous membrane due to abnormalities of anchoring fibrils. Both dominant and recessive DEB have been shown to be caused by mutations in COL7A1, the gene encoding type VII collagen which is the major component of anchoring fibrils. De novo mutation in dominant DEB is rare. In this study, we report a novel de novo glycine substitution mutation in COL7A1 in a Chinese female patient presenting with mild DEB. In search of the mutation, we scanned the entire COL7A1 using polymerase chain reaction (PCR) amplification of all exons of COL7A1, followed by heteroduplex analysis and direct sequencing of the PCR products that exhibited heteroduplex pattern. A G-to-A transition at nucleotide position 6082 within exon 73 of COL7Al was detected. The mutation converted a glycine to an arginine (G2028R) within the triple-helical domain of type VII collagen. It was confirmed that the mutation was present only in the proband. Haplotype analyses suggested that the case arose as a de novo occurrence of autosomal dominant DEB. PMID- 10836610 TI - Three-dimensional comparative study of Merkel cell distribution in various body sites of guinea pigs and rats. AB - The hair cycling pattern differs between humans and mice: for the former it is mosaic, while the latter show a wave pattern. We focused on the presence or absence of synchronization of the hair cycle based on the distribution of Merkel cells. The three-dimensional distribution and number of Merkel cells in the skin at various sites were determined in guinea pigs and rats. Using ethylenediaminetetraacetic acid, various types of hair follicles were obtained from the facial, abdominal, nipple, back, tail, perianal and plantar skin. Epidermal sheets containing numerous hair follicles were immunostained using monoclonal murine antibody CK20 which is generally accepted to be a specific immunocytochemical marker for Merkel cells of various species. Merkel cells were absent in the trunk skin of guinea pigs and rats, in complete contrast to the situation in humans. In vibrissa and other hair follicles within specialized sensitive areas, Merkel cells were found in varying numbers. Some differences in their distribution and number were observed between guinea pigs and rats. The absence of Merkel cells in the hair follicles of the trunk skin of rats and guinea pigs implies that Merkel cells are not essential for the induction and promotion of hair growth but have some different functional significance in hair follicles. PMID- 10836609 TI - The antipsoriatic activity of IL-10 is rather caused by effects on peripheral blood cells than by a direct effect on human keratinocytes. AB - IL-10 is a promising candidate for the treatment of cutaneous disorders. Antipsoriatic efficacy of systemic IL-10 treatment has been already demonstrated. This includes histomorphological changes in the epidermis, suggesting effects on keratinocytes. However, less is known about direct effects of IL-10 on this cell population, although effects are likely since IL-10 receptor expression on keratinocytes has been demonstrated recently. Therefore we analysed the effects of IL-10 on keratinocytes in vitro, using concentrations of human recombinant IL 10 corresponding to those detectable in plasma during therapy. Proliferation, cytokine formation (IL-6, IL-8, IL-1ra), and expression of surface molecules (MHC class I and II, costimulatory molecules CD80 and CD86, CD29, CD54, CD95) were measured in stimulated and unstimulated cells. Although stimulation influenced the expression levels of certain surface markers, no or only slight effects of IL 10 were found. In contrast considerable inhibitory effects of IL-10 on surface molecule expression and cytokine secretion by peripheral blood human monocytes were observed. Our results suggest that the antipsoriatic activity of IL-10 is rather caused by modulatory effects on circulating immune cells, which subsequently might infiltrate the skin, than by direct effects on human keratinocytes. Considering the remarkable antipsoriatic activity of IL-10 and the observation that IL-10 seem to act on peripheral blood mononuclear cells but not on keratinocytes provide further evidence that circulating immune cells play a key role in the pathology of psoriasis. Finally, our results argue against the value of IL-10 therapy in dermatoses strictly limited to keratinocyte involvement. PMID- 10836611 TI - TNF-alpha and serum induce SKALP/elafin gene expression in human keratinocytes by a p38 MAP kinase-dependent pathway. AB - Keratinocytes of inflamed epidermis (psoriasis, wound healing) are hyperproliferative and display an abnormal differentiation programme. This regenerative differentiation pathway is characterized by the induction of genes that are not expressed by keratinocytes in normal skin, such as the cytokeratins CK6, CK16, CK17, and the proteinase inhibitor SKALP/elafin. In the study reported here we investigated the induction and regulation of SKALP expression as a marker for regenerative differentiation in epidermal keratinocytes. Various cytokines and growth factors known to be present in psoriatic epidermis were examined for their ability to induce SKALP gene expression in cultured human keratinocytes. Tumour necrosis factor-alpha (TNF-alpha) and serum were found to be potent inducers of SKALP expression at both the mRNA and the protein levels. SB202190 or SB203580, two specific p38 MAP kinase inhibitors almost completely blocked the induction of SKALP expression by TNF-alpha and serum. These results suggest that in keratinocytes, p38 activity is crucial for the induction of SKALP gene expression. These findings could be relevant for the elucidation of the mechanisms involved in normal and disturbed epidermal differentiation. PMID- 10836612 TI - Alterations of extracellular matrix induced by tobacco smoke extract. AB - Epidemiologic studies have indicated the association between tobacco smoking and skin aging, but the exact mechanism of tobacco smoke-induced premature skin aging is currently unknown. In this study, we investigated the alterations of collagen, matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) in human fibroblasts treated with tobacco smoke extract. Human fibroblasts were exposed to different concentrations of water-soluble extract from tobacco smoke. Human fibroblasts irradiated with ultraviolet A1 (UVA1) were used as positive controls because the mechanism of UVA1-mediated MMP expression has been well characterized. The expression of MMP and TIMP was analyzed semiquantitatively following reverse transcriptase-polymerase chain reaction. Production of type I and type III collagens was detected by Western blotting and biosynthesis of new collagen was assessed by 3H-proline incorporation. Upon treatment with tobacco smoke extract or UVA1 irradiation, the expression of MMP-1 and MMP-3 mRNA was significantly increased in a dose-dependent manner. Maximum induction was observed with 25 microl/ml tobacco smoke extract. In contrast, the expression of TIMP-1 and TIMP-3 mRNA remained unchanged. Western blotting of the supernatant revealed that type I and type III collagens were decreased as compared with untreated controls. Collagen biosynthesis was significantly reduced by 40.1% following treatment with 25 microl/ml tobacco smoke extract. Sodium azide, L-ascorbic acid and Trolox (a water-soluble vitamin E) prevented both the UVA1- and the tobacco-induced alteration of MMP-1. These observations suggest that the imbalance of connective tissue matrix components might contribute to the molecular basis for premature skin aging in smokers. They also suggest that reactive oxygen species including singlet oxygen mediate this process. PMID- 10836613 TI - LAMB3 gene transfection into SV40-transformed keratinocytes from patient with Herlitz junctional epidermolysis bullosa. PMID- 10836614 TI - Symptomatic treatment of respiratory and nutritional failure in amyotrophic lateral sclerosis. AB - Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease characterised by death of upper and lower motor neurones. Nutritional and respiratory failure occurs in most patients with ALS. Nutritional failure occurs primarily as a result of dysphagia, although malnutrition may also develop in the absence of clinically apparent dysphagia. The optimal management of nutrition in early ALS has not been established. In later stages of the disease, parenteral nutritional support using percutaneous endoscopic gastrostomy confers a significant survival benefit in selected patients. Respiratory failure occurs as a result of bulbar, cervical and thoracic loss of motor neurones. Inspiratory muscles are preferentially affected. Management of respiratory failure includes the use of strategies that limit aspiration pneumonia, the reduction in secretions, and positioning of the patient to a maximal mechanical advantage. Use of non-invasive positive pressure ventilation in appropriate patients significantly enhances survival. The decision to undertake invasive mechanical ventilation should be made prior to the development of symptoms that might warrant this intervention. The progressive nature of the condition should be taken into account when such a decision is discussed with the patient and carer. Further studies are required to determine the optimal nutritional requirements of patients with ALS, and to elucidate the physiological changes involved in the decline in respiratory function. PMID- 10836615 TI - Cerebral venous thrombosis: diagnosis and management. AB - In contrast to arterial stroke, cerebral venous thrombosis (CVT) is an infrequent condition which presents with a wide spectrum of signs and with a highly variable mode of onset. The clinician must therefore consider it systematically in all brain syndromes and perform the appropriate neuroimaging investigations: computed tomography (CT) with computed tomography angiography and/or magnetic resonance imaging with magnetic resonance angiography and, if necessary intra-arterial angiography. Once the diagnosis is established, a wide investigation for should be carried out in search of the cause, and treatment started as soon as possible. Treatment is based on the combination of intravenous heparin (followed by oral anticoagulants for 3-6 months), symptomatic treatment (anticonvulsants, analgesics, treatment of increased intracranial pressure) and treatment of the cause. Local thrombolysis is indicated if there is deterioration due to thrombosis extension despite adequate anticoagulation. Diagnosis and treatment of CVT should be considered as an emergency because of the considerable potential for full recovery in this condition. PMID- 10836616 TI - Can we predict flu-like symptoms in patients with multiple sclerosis treated with interferon-beta? AB - Interferon-beta1b treatment in relapsing-remitting multiple sclerosis can frequently induce systemic side effects such as flu-like symptoms with fever. In vitro stimulation of peripheral blood leukocytes with interferon-beta1b before the beginning of therapy shows that patients who develop fever generally have increased levels of interleukin-6. PMID- 10836617 TI - Piracetam in acute stroke: a systematic review. AB - We studied whether the administration of piracetam in acute, presumed ischemic stroke affects case fatality and functional outcome. The Cochrane Stroke Group strategy was used to evaluate all randomized controlled trials of patients with presumed ischemic stroke examined within 48 h; death and (when available) functional outcome were used as end points. Three studies were included; the most recent one contributed more than 97% of the data. There were 501 patients treated with piracetam and 501 controls. Piracetam was associated with a nonsignificant 31% increase in the odds of death (95% CI -5% to 81%). This result was due almost completely to the effect of the larger trial, which, however, reported that the difference in case fatality rate between piracetam and control disappeared after correcting for the imbalance in stroke severity between the two groups. Data on functional outcome were available only for the largest study, and no difference was reported. Data obtained from the manufacturer suggested a nonsignificant trend (-10%) towards reduction in dependency with piracetam (CI -33% to 20%); the proportions of patients dead or dependent in the two groups were the same. Relevant adverse effects were not reported. The evidence from this review does not support routine administration of piracetam in patients with acute ischemic stroke; however, since a possible beneficial effect cannot completely be ruled out, further controlled trials are warranted. PMID- 10836618 TI - Quantitative assessment and correlation of sympathetic, parasympathetic, and afferent small fiber function in peripheral neuropathy. AB - We compared three neurophysiological methods for assessing small nerve fiber function in 40 patients with peripheral neuropathy to determine the various manifestation types of peripheral small fiber neuropathy. Heart rate variation tests were used to assess cardiac parasympathetic small fiber function. Cutaneous vasoconstrictor responses (sympathetic C fibers) induced by deep inspiration were examined with laser Doppler flowmetry. Cutaneous afferent C fiber function was assessed by measurement of axon reflex vasodilatation induced by histamine iontophoresis. All test parameters were significantly lower in patients with peripheral neuropathy than in control subjects. Comparison of the three small fiber systems revealed that functionally different systems are damaged independently, and isolated affection of each fiber type was frequently observed. The three tests are useful noninvasive tools with which to evaluate sympathetic, parasympathetic, and afferent small fiber function in patients with peripheral neuropathy. In many patients functionally different small fiber systems are affected selectively. To diagnose small fiber neuropathy and to evaluate the individual type of manifestation complementary testing of several small somatic and autonomic fiber systems is necessary. PMID- 10836619 TI - Cognitive impairment in systemic lupus erythematosus: a follow-up study. AB - We evaluated outcome and the clinical value of cognitive impairment in systemic lupus erythematosus (SLE). Fifty-one consecutive SLE subjects with or without overt nervous system involvement received two comprehensive neuropsychiatric and neuropsychological assessments, including the Mental Deterioration Battery, the Mini Mental State Examination (MMSE), and tests from the Wechsler Adult Intelligence Scale. The two neuropsychological assessments were made when subjects were in stable neurological condition. Twenty-seven patients were found to have neuropsychiatric symptoms (NP-SLE) at the first assessment, and three others developed them during the follow-up. Fifteen patients (10 NP-SLE) had cognitive impairment at the first assessment. At retest the cognitive deficit persisted in all patients but one (non-NP-SLE) and had developed in four others. In the cognitively impaired subjects scores on MMSE approached the cutoff for an overt dementing condition. No progressively decreasing scores were found on any of the tests. No relationships were shown between neuropsychological diagnosis and neuropsychiatric disorder, neuroradiological findings, disease activity, or steroid and nonsteroid immunosuppressive therapy. Cognitive impairment thus seems to be a stable symptom of CNS involvement in SLE. It corresponds to the subjective complaint of intellectual difficulties and marginal performance on the MMSE. Intellectual deterioration may occur in patients without other symptoms of NP-SLE. Standardized neuropsychological testing methods should be used routinely to assess SLE patients. PMID- 10836620 TI - MRI findings of small subcortical "lacunar-like" infarction resulting from large vessel disease. AB - Small subcortical infarctions resulting from large-vessel disease are often observed. It is important to distinguish these from pure lacunar infarction resulting from small-vessel disease because the investigations and examinations differ. We investigated the differences on brain magnetic resonance imaging (MRI) between small subcortical "lacunar-like" infarcts resulting from large-vessel disease and pure lacunar infarcts. Thirteen subjects with small lacunar-like infarcts (size < 2 cm), resulting from large-vessel disease, and 30 subjects with lacunar infarcts (< 2 cm), without large-vessel disease were studied. We measured infarction size using a 1.5-T MRI device and evaluated silent subcortical hyperintensity lesions using the modified Scheltens' score. Large-vessel lesion was confirmed by conventional angiography, duplex carotid scan, and magnetic resonance angiography. There was no difference in the mean age of the two groups. Cerebrovascular risk factors and atherosclerotic complications were also comparable for the two groups. Progressive stroke was more common in the lacunar like infarction group than in the lacunar infarction group (P = 0.004). Scores for periventricular hyperintensity, white matter hyperintensity, basal ganglia hyperintensity, and total subcortical hyperintensity scores were significantly higher in the lacunar infarction group than in the lacunar-like infarction group. The difference in basal ganglia hyperintensity scores was remarkable (P = 0.001). The enlargement of the perivascular space was also significantly greater in the lacunar infarction group than in the lacunar-like infarction group. These findings seem to reflect differences in the pathogenesis of infarction between the two groups. Silent subcortical hyperintensity lesions and enlargement of perivascular space are useful for between distinguishing small lacunar-like infarct resulting from large-vessel disease and pure lacunar infarction. This may have significant implications for the management of patients with lacunar-sized infarctions. It suggests that the pathogenesis of lacunar-sized infarction is variable. PMID- 10836621 TI - Validity and reliability of two muscle strength scores commonly used as endpoints in assessing treatment of myasthenia gravis. AB - Valid and reliable measurements of muscle impairment are needed to assess therapeutic efficacy in patients with generalized myasthenia gravis (MG). In 22 patients we compared the validity and interobserver reliability of two scoring methods commonly used as main endpoints in clinical trials, i.e., the Myasthenic Muscle Score (MMS) ranging from 0 to 100 (normal) and the Quantified Myasthenia Gravis Strength Score (QMGSS) ranging from 0 (normal) to 39. Each score is correlated more with functional scale and less with the patient's self evaluation. Using intraclass correlation we found strong agreement between observers for both the MMS (r = 0.906) and the QMGSS (r = 0.905). The correlation between MMS and QGMSS was high (r = 0.87). The reliability of neither score depended on any specific item, since the removal of individual items did not significantly alter the intraclass correlation coefficient (ranging from 0.86 to 0.93). PMID- 10836622 TI - Carotid atherosclerosis and cerebral white matter lesions in a population based magnetic resonance imaging study. AB - Cerebral white matter lesions are frequently observed on magnetic resonance imaging of elderly, nondemented persons. There is evidence that white matter lesions are involved in the pathophysiology of cognitive decline and dementia. White matter lesions can be divided into those in the periventricular and those in the subcortical region. Pathological and epidemiological studies suggest that atherosclerosis is involved in the pathogenesis of these lesions. Our study reports on the association between atherosclerosis in the carotid arteries and white matter lesions in a population-based study among 1077 elderly subjects. We randomly sampled 1077 subjects aged between 60-90 years from two prospective population-based studies. All subjects underwent ultrasonography of the carotid artery. In addition, 1.5 T magnetic resonance imaging was performed; white matter lesions in the subcortical and periventricular regions were rated separately. With increasing number of plaques in the carotid artery the severity of periventricular white matter lesions increased (Ptrend = 0.03), but not the severity of subcortical white matter lesions (Ptrend = 0.19). In addition, an increase in intima media thickness was borderline significantly associated with an increased severity of periventricular white matter lesions (Ptrend = 0.09), but not of subcortical white matter lesions (Ptrend = 0.68). These findings suggest that partly dissimilar pathogenetic mechanisms are involved in the etiology of periventricular and subcortical white matter lesions. PMID- 10836623 TI - A role for preirradiation PCV chemotherapy for oligodendroglial brain tumors. AB - Oligodendroglial tumors have been identified as a subgroup of glial neoplasms with a distinctly better response to chemotherapy and overall survival than purely astrocytic gliomas. Here we report our experience with adjuvant postirradiation and preirradiation chemotherapy using procarbazine, lomustine, and vincristine (PCV) in 27 patients with WHO grade II or III oligodendroglioma or oligoastrocytoma. The efficacy of chemotherapy was assessed according to the Macdonald response criteria (complete response, CR; partial response, PR; stable disease, SD; progressive disease, PD) and progression-free survival intervals by computed tomography or magnetic resonance imaging. First, we confirm that PCV salvage therapy for patients progressing after radiotherapy is highly effective (n = 11, 1 CR, 5 PR, 5 SD; median progression-free survival has not yet been reached, but is longer than 18 months). Second, 3 patients who received radiotherapy plus PCV as first-line therapy achieved CR and 2 achieved SD, and all 5 are progression-free with a median follow-up of 12 months. Third, given these encouraging results, 11 patients received postoperative preirradiation PCV chemotherapy and were given radiotherapy only upon progression. Preirradiation PCV chemotherapy was also effective (2 CR, 3 PR, 6 SD; median progression-free survival has not been yet reached, but is longer than 14 months). Patients with anaplastic oligoastrocytomas were as likely to respond to PCV chemotherapy, as were patients with anaplastic oligodendroglioma. Three patients who had previously responded to PCV were successfully treated with a second course of PCV upon recurrence. PCV chemotherapy was also effective in patients with leptomeningeal spread of oligodendrogliomas. A randomized prospective trial is required to compare the effectiveness and neurotoxicity of first-line PCV chemotherapy followed by radiotherapy to the traditional reverse sequence. PMID- 10836624 TI - Visually induced seizure caused by "opto-acoustic relaxation system". PMID- 10836625 TI - Multiple secondary Leber's hereditary optic neuropathy mutations in Italian patients with multiple sclerosis. PMID- 10836627 TI - An early description of striatonigral degeneration. PMID- 10836626 TI - Contrast enhancement in progressive multifocal leukoencephalopathy: a predictive factor for long-term survival? PMID- 10836628 TI - Damage to peripheral nerves following radiotherapy at the wrist. PMID- 10836629 TI - Raymond Garcin (1897-1971). PMID- 10836630 TI - Beta-sheet breaker peptides reverse conformation of pathogenic prion proteins. PMID- 10836631 TI - Position sense during imperceptibly slow movements. AB - Proprioceptive input provides the nervous system with information related to body position and movement. This study investigated how normal humans perceive information related to the position of a limb when it is either at rest or moving very slowly, below the threshold for movement perception. Each subject's left hand, hidden from view, was horizontally translated in the frontal plane such that joint rotation was largely isolated to the shoulder. The translation speed was too slow for the subjects to detect movement. Subjects indicated their perception of hand position at 1- or 2-min intervals by pointing with the right index finger to the perceived location of the tip of the left middle finger. The constant error (i.e., bias) and variable error (i.e., SD of mean constant error) of each pointing movement was quantified. Partway through the trial, the direction of hand movement was reversed. In two additional experimental conditions, the subjects pointed at 1- or 2-min intervals at the stationary left hand, either when they assumed the hand would be moved or when they knew it would not. During slow movement, errors in the frontal and sagittal planes were found to be independent, and therefore, data analysis focused on the frontal plane, as this was the plane relevant to the subjects' task. All subjects performed the task sufficiently well to demonstrate a clear perception of hand location during the slow movement. The accuracy of perception was better when the left (target) hand was in its ipsilateral hemifield and, correspondingly, when the right hand pointed to its contralateral hemifield. There was no significant difference in constant error when the hand moved slowly, although there was a slightly higher variable error during slow movement than when the hand was stationary. Based on the similarity of results in trials with very slow and no hand movement, it was concluded that position-sense percepts are more accurately distinguished by the speed of movement rather than whether movement is occurring or not. PMID- 10836632 TI - A second look at the barriers of the medial basal hypothalamus. AB - The cell bodies of hypothalamic secretory neurons are localized in areas protected by the blood-brain barrier (BBB), whereas their axon terminals are localized in the median eminence, which lacks a BBB. This implies a complex barrier system, allowing neurons of the central nervous system to secrete into the blood stream without making the BBB leaky. In the present study, three experimental protocols were applied to clarify certain relevant aspects of the barriers operating in the medial basal hypothalamus of the rat. We established that the milieu of the arcuate nucleus is exposed to both the ventricular and the subarachnoidal cerebrospinal fluid (CSF). The median eminence milieu, the perivascular space of the portal vessels, and the subarachnoid space appear to be in open communication; also, beta2-tanycytes establish an efficient barrier between the median eminence milieu and the ventricular CSF. Similarly, beta1 tanycytes establish a lateral barrier, separating the intercellular space of the median eminence from that of the arcuate nucleus. We also found that the glucose transporter I (GLUT I), a BBB marker, is localized throughout the whole plasma membrane of beta1-tanycytes, but is missing from beta2-tanycytes. Expression of GLUT I by tanycytes progressively develops during the first postnatal weeks; while the degree of damage of the arcuate nucleus by administration of monosodium glutamate, at different postnatal intervals, parallels that of the GLUT I immunoreactivity of beta1-tanycytes. An explanation is offered for the selective destruction of the arcuate neurons by the parenteral administration of monosodium glutamate to infant rats. PMID- 10836633 TI - Recovery of hindlimb motor functions after spinal cord transection is enhanced by grafts of the embryonic raphe nuclei. AB - In this study, a piece of embryonic tissue from the raphe nucleus was transplanted into the spinal cord below the lesion 1 month after transection. Two months later the recovery of hindlimb motor function in rats which had received a transplant of neural tissue (ST rats) was much better than in spinal control animals without the graft (SC rats). Analysis of the electromyographic (EMG) activity showed that the timing of muscle activity during locomotor-like movement of hindlimbs in ST rats was more regular than in SC rats. In SC rats the relationships between EMG burst duration (soleus, tibialis anterior) and step cycle duration were significantly altered. The restoration of hindlimb motor function of ST rats was also reflected in the better interlimb coordination during locomotor-like hindlimb movements. The results of several behavioural tests demonstrated that the responses to stimulation of various receptors, such as tactile or proprioceptive, in ST rats were more complex than in SC rats. Additionally, unlike in SC animals, in ST rats long-lasting spontaneous episodes of air stepping movement of hindlimbs accompanied by a relatively high amplitude of EMG activity were obtained. These results confirm that grafted embryonic raphe nuclei which contain serotoninergic cells are likely to increase the excitability of neuronal circuitry in the injured spinal cord. Moreover, transplantation of embryonic raphe nuclei encourages the recovery of hindlimb motor function in adult rats even when the grafting is carried out several weeks after spinal cord injury. PMID- 10836634 TI - Extent of compensation for variations in monkey saccadic eye movements. AB - We investigated and quantified the ability of the primate saccadic system to generate accurate eye movements in spite of naturally occurring variations in saccadic speed and trajectory. We show that the amplitude of a series of saccades directed to the same target is positively correlated to their peak speed, i.e., the faster the saccade, the bigger its amplitude. We demonstrate that this result cannot be simply accounted for by the main sequence, and that on average the saccadic system is able to compensate for only 61% of the variability in speed. Deviations from the average trajectory are also only partially compensated: the underlying mechanism, which tends to bring the eyes back toward the desired trajectory, underperforms for small movements and overperforms for large movements. We also demonstrate that the performance of this compensatory mechanism, and the metrics of saccades in general, do not depend on the presence of visual information during the movement. By showing that deviations from the desired behavior are corrected during the saccade, our results further support the hypothesis that the innervation signal that generates saccadic eye movements is not pre-programmed but rather is dynamically adjusted during the movement. However, the compensation for deviations from the desired behavior is only partial, and the underlying mechanisms have yet to be completely understood. Although none of the current models of the saccadic system can account for our results, some of them, if appropriately modified, probably could. PMID- 10836635 TI - Moving weightless objects. Grip force control during microgravity. AB - When we move grasped objects, our grip force precisely anticipates gravitational and inertial loads. We analysed the control of grip forces during very substantial load changes induced by parabolic flights. During these flight manoeuvres, the gravity varies between hypergravity associated with a doubling of normal terrestrial gravity and a 20-s period of microgravity. Accordingly, the contribution of the object's weight to the load changed from being twice the normal value to being absent. Two subjects continuously performed vertical and horizontal movements of an object equipped with grip force and acceleration sensors. Whereas, during vertical movements performed under normal and hypergravity, a load force maximum occurred at the lower turning point and a minimum at the upper turning point, the load force pattern was completely changed under microgravity. In particular, the upper turning point was also associated with a load force maximum. Analysis of the grip forces produced by the two subjects revealed that the grip forces underwent the same characteristic changes as the load forces. Thus, subjects were able to adjust grip forces in anticipation of arm movement-induced fluctuations in load force under different and novel load conditions. Adaptation to changing levels of gravity was also obvious when the vertical and horizontal movements were compared: grip forces depended heavily on movement direction during normal and hypergravity but not during microgravity. The predictive coupling of grip force and load force was observed even during transitions between gravity levels, indicating rapid adaptation to changing load conditions. To account for the striking preservation of the normal characteristics of grip force control, we suggest that a highly automatized, extremely flexible sensorimotor mechanism firmly implemented within the central nervous system can cope with even massive changes in the environmental conditions. PMID- 10836636 TI - Modulation of stretch-evoked reflexes in single motor units in human masseter muscle by experimental pain. AB - The interaction between muscle pain and motor function of the jaw has been examined in recent years, but the nature of the modulation of the short-latency stretch reflex by pain is not fully understood. In this study, the reflex responses to stretch were measured in single low-threshold motor units that were kept discharging at a constant frequency, before, during and after the induction of experimental pain in one masseter muscle by controlled infusion of hypertonic saline. The probability of evoking a reflex response in individual motor units in the painful muscle at near-monosynaptic latency was reduced by a mean of about 20%. However, the overall reflex response in the surface electromyogram of both the ipsi- and contralateral masseter muscles was greater during pain. This was apparently a secondary response to the pain-induced increase in pre-stimulus activity in the motoneurone pools of both muscles, because increased motoneurone excitability may facilitate stretch reflexes. It is concluded that the most likely explanation for the reduced reflex response of low-threshold masseter motor units during experimental pain is a tonic reduction in the fusimotor drive to the masseter spindles. PMID- 10836637 TI - Stimulation in the rostral pole of monkey superior colliculus: effects on vergence eye movements. AB - Recent studies have indicated that the superior colliculus (SC), traditionally considered to be saccade-related, may play a role in the coding of eye movements in both direction and depth. Similarly, it has been suggested that omnidirectional pause neurons are not only involved in the initiation of saccades, but can also modulate vergence eye movements. These new developments provide a challenge for current oculomotor models that attempt to describe saccade-vergence coordination and the neural mechanisms that may be involved. In this paper, we have attempted to study these aspects further by investigating the role of the rostral pole of the SC in the control of vergence eye movements. It is well-known that, by applying long-duration electrical stimulation to rostral sites in the monkey SC, saccadic responses can be prevented and interrupted. We have made use of these properties to extend this paradigm to eye movements that contain a substantial depth component. We found that electrical intervention in the rostral region also has a clear effect on vergence. For an eye movement to a near target, stimulation leads to a significant suppression and change in dynamics of the pure vergence response during the period of stimulation, but the depth component cannot be prevented entirely. When these paradigms are implemented for 3D refixations, the saccade is inactivated, as expected, while the vergence component is often suppressed more than in the case of the pure vergence. The data lead us to conclude that the rostral SC, presumably indirectly via connections with the pause neurons, can affect vergence control for both pure vergence and combined 3D responses. Suppression of the depth component is incomplete, in contrast to the directional movement, and is often different in magnitude for 3D refixations and pure vergence responses. The results are discussed in connection with current models for saccade-vergence interaction. PMID- 10836638 TI - Do anticipatory postural adjustments occurring in different segments of the postural chain follow the same organisational rule for different task movement velocities, independently of the inertial load value? AB - Dynamic phenomena, termed anticipatory postural adjustments (APA), are known to precede the onset of voluntary movement. Their anticipatory nature confers a particular status on APAs: as they cannot be triggered reflexly by afferent signals induced by a voluntary movement, it can be asked whether the APA parameters are centrally programmed as a function of some task movement parameters or are only the peripheral consequence of control variables. To this end the present study aims to determine whether the APAs occurring at the different sites of the postural chain yield the same accelerometric patterns and follow the same organisational rules when the task movement velocity changes, independently of the inertial load value. Subjects performed unilateral shoulder flexions at maximal and submaximal velocities, with (IUF) and without (OUF) an additional inertial load. Accelerometers were attached to the wrist and trunk, and on both sides of the body at shank, thigh, hip and shoulder. The results show that: 1) there was evidence of anticipatory acceleration in all segments of the postural chain; 2) each acceleration profile for the anticipatory phase was maintained over different focal movement velocities whether there was an additional load or not; 3) there were significant linear relationships between the amplitude of each segmental anticipatory acceleration and the square of the task movement velocity, the slope of which differs as a function of the inertial load; 4) there were close intersegmental correlations between these anticipatory accelerations which did not depend on the inertial load. In addition the correlation between the lower limbs and the opposite side of the body was positive, suggesting a diagonal postural pattern. A comparison of the present kinematic data with the corresponding EMG data reported in the literature argues in favour of a centrally determined kinematic pattern. It is proposed that the diagonal postural pattern between postural segments be considered as one of the order rules which could simplify the control process of asymmetrical movement. The anticipatory kinematics of each of the body segments would be calibrated with the velocity and the inertial load and scaled to the other segments to counteract the perturbing effect of the asymmetrical focal movement on body balance. PMID- 10836639 TI - Neuronal connections in the medial geniculate body of the guinea-pig. AB - The spontaneous and evoked activities of individual pairs of single units were recorded simultaneously with the same microelectrode in the medial geniculate body (MGB) of ketamine-xylazine-anaesthetised guinea-pigs. Cross-correlograms (CCGs) of spike train pairs were computed and divided on the basis of correlation peak shape into four classes [a unilateral narrow (UN) peak, a centrally positioned wide (CW) peak, a complex peak and no significant peak] interpreted in terms of the functional connection between neighbouring neurones. The shift predictor procedure was applied with the aim of removing the effect of the stimulus on the final CCG shape. The occurrence of correlation peak types and the distribution of correlation coefficients were found to be similar for the spontaneous activity during silent periods following acoustical stimulation and for the long-lasting recording of spontaneous activity. CCGs in 38% of pairs computed during silent interstimulus intervals contained a UN peak, suggesting a monosynaptic excitatory connection. Almost 20% of all pairs expressed a CCG shape typical for a common input, i.e. a CW peak. In 5% of cases multiple, so-called complex peaks, were found. About 20% of the CCGs contained no significant correlation peak in the interstimulus period, which is typical for a very weak or absent functional connection between recorded neurones. No inhibitory interaction (groove in the CCGs) between recorded pairs was observed. The distribution of correlation peak shapes was similar when calculated during acoustical stimulation and during silent interstimulus intervals. CCGs computed during presentation of four acoustical stimuli (pure tone bursts, noise bursts, natural call whistle and artificially inverted whistle) showed most frequently a UN peak (28-37%) followed by CCGs with no significant peak (18-28%) and with a UN/CW peak (14-23%). On average, the occurrence of UN peaks tended to be less frequent during stimulus presentation than in silent conditions, but the difference was not statistically significant. The most frequent occurrence of clear UN peaks was found in the medial part of the MGB (from 52-64% of pairs depending on the type of acoustical stimulus), while the least was observed in the ventral part of the MGB (12-22%). In contrast, CW peaks were most frequently expressed in pairs located in the ventral part of the MGB (18-33%), while neuronal pairs in the medial part revealed a very low occurrence of CW peaks (0-7%). The occurrence of independently firing neurones was lowest in the medial part of the MGB (8-20% of pairs) in comparison with the ventral (31-39%) and dorsal (12-41%) parts. In 20% of pairs acoustical stimulation produced a change in the type of correlation peak present during spontaneous activity. Most frequently, a CW peak (shared input) changed to a flat CCG, which represents independently firing neurones. In some pairs higher connection strengths (as expressed by the value of the correlation coefficient) were found for silent interstimulus intervals than for acoustical stimulation. This suggests that in the MGB the stimulus may desynchronise the spontaneous activity of simultaneously firing units in neuronal pairs. PMID- 10836640 TI - Frontal plane dynamic stability and coordination in subjects with cerebellar degeneration. AB - Objective attempts to characterize postural control in subjects with cerebellar (Cb) pathology have focused primarily on sagittal plane responses to static standing, semi-dynamic standing, and platform perturbation. Repeated, dynamic, functional movement may provide a better opportunity to study the effects of ataxia on frontal plane postural stability. The purpose of this investigation was to quantify lateral stability using center of gravity (CG) and center of pressure (CP) movement analysis, and to examine motor responses necessary to complete a repetitive stepping task. Whole body kinematic and kinetic data were collected as patients with Cb degeneration and non-disabled volunteers repeatedly ascended forward and descended backward from a 7.6-cm step to the beat of a metronome. Cb subjects demonstrated significantly greater lateral CG instability, consistent with increased CG velocity and displacement variability in the frontal plane compared to non-disabled subjects. Significantly greater lateral CP displacement was found in subjects with Cb degeneration. Phase plot patterns from patients with Cb degeneration showed poor movement quality, indicated by abruptly changing CG phase plot patterns and abnormal CG acceleration and deceleration phases. Ataxic postural reactions were observed consistently during lateral weight shifting phases toward the stance limb. We hypothesized that lateral postural instability in Cb subjects stems from altered lower extremity intersegmental coordination and inadequate lateral CG velocity control. Frontal plane CG instability may necessitate a wide-based gait. Despite the varied neuroanatomic sites of degenerative Cb pathology, subjects with Cb pathology demonstrated similar CG movement patterns and altered motor strategies to avoid destabilization. Dynamic assessment provides important information regarding frontal plane instability not revealed by static assessment methods. These findings suggest that a dynamic, constrained stepping task effectively differentiates Cb and non-disabled volunteers, contributing to our knowledge of the effect of ataxic movement disturbances on frontal plane postural stability and lower limb coordination. PMID- 10836641 TI - The role of proprioception and attention in a visuomotor adaptation task. AB - The role of proprioception in the control and adaptation of visuomotor relationships is still unclear. We have studied a deafferented subject, IW, and control subjects in a task in which they used single joint elbow extension to move to a visual target, with visual feedback of the terminal position provided by a cursor displayed in the plane of their movements. We report the differences in movement accuracy between the deafferented subject and controls in the normal task and when challenged with a cognitive load, counting backwards. All subjects were less accurate when counting; this was a small effect for the controls (<10% change) but much greater for the deafferented subject (>60% change). We also examined changes in movement kinematics when the instructed amplitude was altered via a changed gain between final arm position and presentation of the feedback cursor. The deafferented subject maintained temporal movement parameters stable and altered amplitude by scaling force (i.e. changed peak velocity), whereas the controls scaled both movement velocity and duration. Finally, we compared the subjects' adaptation of movement amplitude after a period of exposure to the changed visuomotor gain. The deafferented subject was able to adapt, but his adaptation was severely impaired by the counting task. These results suggest that proprioception is not an absolute requirement for adaptation to occur. Instead, proprioception has a more subtle role to play in the adjustment to visuomotor perturbations. It has an important role in the control of reaching movements, while in the absence of proprioception, attention appears necessary to monitor movements. PMID- 10836642 TI - Interhemispheric transfer of visual motion information after a posterior callosal lesion: a neuropsychological and fMRI study. AB - Interhemispheric transfer of visual information was investigated behaviourally and with functional magnetic resonance imaging (fMRI) 6 months after a lesion of the posterior two-thirds of the corpus callosum. On tachistoscopical left hemifield presentation, the patient was severely impaired in reading letters, words and geographical names and moderately impaired in naming pictures and colours. In contrast, interhemispheric transfer of visual motion information, tested by verbal report of the direction of short sequences of coherent dot motion presented within the left hemifield, was preserved. The pattern of cerebral activation elicited by apparent motion stimuli was studied with fMRI and compared to that of normal subjects. In normal subjects, apparent motion stimuli, as compared to darkness, activated strongly striate and extrastriate cortex. When presented to one hemifield only, the contralateral calcarine region was activated while regions on the occipital convexity, including putative area V5, were activated bilaterally. A similar activation pattern was found in the patient with a posterior callosal lesion; unilateral left or right hemifield stimulation was accompanied by activation in the contralateral and ipsilateral occipital convexity. Ipsilateral hemifield representation in the extrastriate visual cortex is believed to depend on callosal input. Our observation suggests that this is not the case for visual motion representation and that other, probably parallel, pathways may mediate visual motion transfer after posterior callosotomy. PMID- 10836643 TI - Age-dependent response of the mouse barrel cortex to sensory deprivation: a 2 deoxyglucose study. AB - The effect of age on the plastic response of vibrissal barrel cortex to deprivation was examined in adolescent (1 month at the start of the procedure, 2 months at testing) and mature (10- to 11-month-old) mice. A single vibrissa was plucked out for 3 weeks and allowed to regrow for 10 days; it was previously found that this deprivation paradigm induces strong downregulation of the deprived input. The results of deprivation were assessed with 2-deoxyglucose functional brain-mapping autoradiography. Deprivation was found to reduce the ability of the deprived vibrissa to activate the cortex both in adolescent and mature mice. However, while in young animals the decrease of the extent of cortical labeling, compared with the normal control, was observed in all examined cortical layers (II/III, IV, and V), in older mice the effect was reduced in layers II/III and absent in layer IV. The suppression of response of the infragranular layers was not affected by age. Transition from adolescent to mature adulthood brings about a layer-specific decline in depression of the cortical response to the deprived input. PMID- 10836644 TI - Liver transplantation as rescue treatment in a patient with primary AL kappa amyloidosis. AB - Although involvement of the liver is common in systemic amyloidosis, clinical manifestations of hepatic dysfunction and liver biochemical abnormalities are often absent or only mild. Here we report on a patient with primary amyloidosis and rapid development of liver failure, who was successfully treated by liver transplantation. The patient is a 61-year-old Swedish man who was admitted to the local hospital for spontaneous rupture of the spleen. Before admission, he had suffered from diffuse upper abdominal discomfort, diminished appetite, and had lost 15 kg in 6 months. Shortly after splenectomy, he developed cholestatic liver failure with moderate hepatomegaly, jaundice, ascites and hyponatremia. Over a period of 3 weeks his liver failure progressed, renal function deteriorated rapidly, and he developed encephalopathy. Liver transplantation was performed on the 35th day after splenic rupture. Histological examination revealed extensive deposits of amyloid in the spleen and liver. N-terminal amino acid sequence analysis of the amyloid protein, purified from the patient's native liver, revealed an AL protein of kappa I-type origin. The postoperative course was uncomplicated, apart from one episode of sepsis and one course of treatment for acute rejection. He was discharged from hospital with normal liver function and good kidney function. One year after surgery, he was in good condition, with normal liver function. However, a liver biopsy taken at the same time showed de novo amyloid deposits in the grafted liver. We conclude that liver transplantation may be indicated as a life-saving procedure in rapidly progressing hepatic amyloidosis with cholestatic jaundice, although the underlying disease has not changed. PMID- 10836645 TI - Physiologic and metabolic results of pancreatic cold storage with Histidine Tryptophan-Ketoglutarate-HTK solution (Custodiol) in the porcine autotransplantation model. AB - Delayed graft function (DEGF) remains an obscure phenomenon in organ transplantation. For the optimal washing of the compounds of the different organ flush solutions, adequate temperature and equilibrium of electrolytes have to be provided. A total of 29 land-race pigs weighing 37.3-5.4 kg were included in this study. According to the model, the left hemipancreas was perfused with Histidine Tryptophan-Ketoglutarate (HTK)-solution and autotransplanted after 24 h (G1, n = 13) and 48 h (G2, n = 4) of cold storage (CS). Results were compared with grafts perfused with UW-solution and autotransplanted after 24 h (G3, n = 8) and 48 h (G4, n = 4) CS respectively. Daily measurements of glycemia, glucosuria, amylase and lipase were carried out. HTK perfusion resulted in an increase in wet weight of the grafts after 24 h and 48 h CS (P = 0.031 vs UW). Postoperative glycemia levels in pancreases flushed with HTK-solution were higher after 48 h than after 24 h CS until the 6th postoperative day, when the glycemia returned to normal range (P = 0.02), suggesting a delayed endocrine graft function. The mean IVGTT values attained after full function were comparable in G1 and in G3 (-1.22 +/- 0.23 vs. -1.5 +/- 0.65). The rises in serum amylase and lipase levels were more pronounced after 48 h CS in both HTK and UW groups, (P = n.s.). Appearance of interstitial and intracellular edema after CS and reperfusion did not influence the function. CONCLUSION: HTK-solution is suitable for 24 h pancreatic preservation in vivo; the perfusion requires at least 4 min for electrolyte equilibration. Long preservation time (48 h) resulted in a transitory DEGF. PMID- 10836646 TI - Altered expression of matrix metalloproteinases and tissue inhibitors of metalloproteinases in acutely rejected myocardium and coronary arteriosclerosis in cardiac allografts of nonhuman primates. AB - Matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) are important in any process of tissue remodeling. However, there is no report evaluating their expression in cardiac allografts in human or non-human primates. Heterotopic cardiac transplantation was performed on Japanese monkeys. Subjects were treated with chimeric anti-human lymphocyte function-associated antigen-1 monoclonal antibody for 2 weeks. Heart grafts were harvested at days 1 95 (n = 7). Native monkey hearts were used as controls (n = 2). We examined expression of MMP-2, MMP-9, TIMP-1, and TIMP-2 using immunohistochemistry and in situ reverse transcriptase polymerase chain reaction (RT-PCR). In the myocardium, the expression of MMP-2 was increased in the spindle-shaped cells of acutely rejected myocardial interstitium and prior to the presence of mononuclear cell infiltration at days 1-41. TIMP-1 and 2 expression was enhanced in association with the progression of fibrosis at days 40-95. In the coronary arteries of chronically rejected allografts, enhanced MMP and decreased TIMP expression was observed in both thickened intima and media at days 40-95. The medial MMP mRNA expression was observed before the development of intimal thickening occurred at days 7-28. MMPs are critical for the progression of acute and chronic rejection, and TIMP predominance plays important roles in fibrosis in association with acute rejection. Expression of MMPs and TIMPs is a sensitive indicator of acute and chronic cardiac rejection. PMID- 10836647 TI - Polymorphonuclear leukocyte functions as predictive markers for infections after organ transplantation. AB - Infectious complications are still a major cause of morbidity and mortality after organ transplantation, and early therapy would certainly reduce the risk associated with severe infections. We therefore investigated the significance of polymorphonuclear leukocyte (PMN) functional tests as predictive markers for infection in transplant patients under immunosuppressive therapy in a longitudinal study. In 41 patients, blood PMN migration and reactive oxygen species release, the blood levels of PMN elastase, malondialdehyde, neopterin, sICAM-1 and sVCAM-1, and urine neopterine were measured in 3- and 4-day intervals after liver-, kidney-, kidney-pancreas-, and heart and lung transplantation. PMN migration was determined in whole blood and estimated by the amount of PMNs to penetrate into a membrane filter upon FMLP stimulation. Three groups of patients were formed according to their postoperative course. Group I patients (n = 23) had no or only minor local infection, group II patients (n = 11) had infections with distinct systemic involvement, and group III patients (n = 7) developed sepsis. A first elastase-level of over 100 mg/L after surgery, followed by a drop in the amount of blood PMNs ready to migrate, on FMLP stimulation, to below 12 %, turned out to be a marker for impending infection, whereas all other parameters tested were not predictive. In six of seven group III patients, this marker became positive (sensitivity 85.7 %) up to 15 days before clinical manifestation of sepsis. In group I (largely uneventful recovery) only one of 23 patients was positive (specificity 95.6 % ), whereas group II patients were in between (4 of 11 positive). By this method it seems possible to diagnose severe infections in the pre-clinical phase, which may help prevent them if treatment is begun promptly. PMID- 10836648 TI - Long-term results of 1047 cadaveric kidney transplantations with special emphasis on initial graft function and rejection. AB - We studied the effect of initial graft function and acute rejection on graft survival in 1047 cadaveric renal transplantations during 1991-1997 with a constant policy of donor selection, graft allocation, and immunosuppression. The overall 1- and 5-year patient survival rates were 96 % and 88 %, and the 1- and 5 year graft survival (GS) rates were 92 % and 78 %. Delayed graft function (DGF) occurred in 31 % and there were 1.2 % never-functioning grafts. One-year GS in transplantations with early graft function (EGF) was 95 % compared to 87 % in DGF (P < 0.001). Donor age and cause of death, type of graft perfusion and cold ischemia time, and type and length of dialysis treatment were significant factors in determining the onset of graft function. These factors did not have a significant direct effect on GS. Early ( < 100 days) acute rejection occurred in 25 %. In transplantations without rejection, the 1 and 5-year GS was 93.3 % and 80.8 %. In acute rejection responding to steroids, the GS was equal to that up to 3 years, but after that a significantly worse survival rate was observed (1- and 5-year GS: 93.6 % and 73.4 %). DGF was detrimental to GS both in transplantations without rejection and in all rejection types. PMID- 10836649 TI - General compliance after liver transplantation for alcoholic cirrhosis. AB - In recent years, alcoholic cirrhosis has been accepted as an indication for OLT, compliance of patients suffering from alcoholic cirrhosis is still under discussion, however. 118 patients who had undergone OLT for alcoholic cirrhosis were considered for analysis. The mean follow-up time of the study population was 53.7 +/- 38.9 months. Compliance was defined by 3 parameters: 1. Sobriety. Fifteen (13%) out of 118 recipients suffered an alcohol relapse during the observation period. There was no difference between the groups with or without alcohol relapse concerning compliance with medication, incidence of rejection, or adherence to check-ups. 2. Drug-compliance. Nineteen recipients (16 %) were not within the target range with the immunosuppressive medication. Comparison of the compliant- and non-compliant groups produced a significant difference for late acute rejection, the other parameters being similar in the subgroups. 3. Adherence to appointments. Nearly all patients in the study population ( > 95 %) were compliant with both transplant and psychological appointments in the outpatient clinic. In conclusion, analysis of our data indicates that patients with OLT for alcoholic cirrhosis are compliant, although alcohol relapse occurs in 13 % of recipients. PMID- 10836650 TI - The efficacy of adjuvant cytostatic therapy after organ transplantation for malignancy: an experimental study with a combined transplantation/tumor model. AB - New data show that perioperative cytostatic therapy is beneficial in the case of liver transplantation for hepatic cancer. However, it has not been established clearly whether chemotherapy interferes with graft rejection. We therefore studied the interactions between tumor growth and graft rejection, especially with regard to chemotherapy, using a combined tumor/transplantation model. As a tumor model, we used the Novikoff hepatoma, a malignant hepatoma that was injected subcutaneously into the backs of rats. Heterotopic heart grafting served as the transplantation model. In a first step (a), we studied the effect of cytostatic therapy on tumor growth: tumor cells were injected, and in four groups epirubicin, cyclosporine, epirubicin + cyclosporine, and placebo were applied, in corresponding groups, transplantation was additionally performed. Tumor growth was measured and the resected tumors were examined by histology and immunohistology. In a second step (b), we studied the effect of chemotherapy on graft rejection: transplantation was performed and the above-mentioned drugs were applied; in corresponding groups, a solid tumor was additionally induced and resected immediately before transplantation. The results of these procedures were as follows: (a) Epirubicin decreased tumor growth and diminished the volume increasing effect of cyclosporine significantly. After transplantation, tumor growth was similar. (b) Epirubicin prolonged graft survival significantly, and the combination with cyclosporine had an augmenting effect. In the corresponding groups, graft survival was similar. In conclusions. chemotherapy diminishes the tumor-increasing effect of cyclosporine and does not interfere negatively with graft survival. It might therefore be beneficial after transplantation for malignancy. PMID- 10836651 TI - Efficacy of Daclizumab in an African-American and Hispanic renal transplant population. AB - Current immunosuppressive regimens have decreased acute rejection rates during the 1st year after renal transplantation. However, this decrease has not been as marked in high-risk groups, such as African-American and Hispanic renal transplant recipients. We compared two simultaneous cohorts of altogether 36 African-American and Hispanic renal transplant recipients. Cohort one received a regimen of mycophenolate mofetil, prednisone, and a calcineurin inhibitor. The second cohort received the same protocol with the addition of Daclizumab (1 mg/kg for five doses given every 2 weeks). The median follow-up was 15.2 months (range 11.8-19.9 months). One patient in the Daclizumab-treated group and seven patients in the control group experienced an acute rejection episode. The rejection-free survival was significantly higher in the Daclizumab-treated group (94.4 %) as compared to the control group (66.7 %, Log-rank < 0.05) at 17 months after transplantation. A Cox Proportional Hazard model revealed lack of Daclizumab therapy as the only significant risk factor for acute rejection. (hazard ratio 7.0, 95 % CI = 1.1-48). The addition of the IL-2 receptor blocker Daclizumab to a triple therapy regimen may decrease early acute rejection in the high-risk groups of African-American and Hispanic patients. PMID- 10836652 TI - Heparin interference with reverse transcriptase polymerase chain reaction of RNA extracted from lungs after ischemia-reperfusion. AB - The reverse transcriptase polymerase chain reaction (RT-PCR) is a rapid and sensitive method for detecting gene expression. However, when we used this technique to study gene expression of cytokines in ischemic and ex-vivo reperfused rat lungs as a model for transplantation, significant inhibition of RT PCR reaction was observed. To optimize RT-PCR conditions, RNA was extracted from rat lungs after flushing, preservation, and reperfusion. RNA was further purified and PCR conditions were modified with various strategies. We found that heparinase I pretreatment completely overcame the inhibitory effects of RT-PCR using RNA extracted from lung tissues after ischemia-reperfusion. With this treatment, a dramatic increase in tumor necrosis factor-a (TNF-a) mRNA was revealed from lung tissues after ischemia-reperfusion. This result suggests that residual heparin in lung tissue interferes with RT-PCR. Because heparinization is routinely used during clinical and experimental organ transplantation, we recommend the treatment of RNA samples with heparinase prior to RT-PCR. PMID- 10836653 TI - Daclizumab prevents acute rejection and improves patient survival post transplantation: 1 year pooled analysis. AB - Daclizumab is a genetically engineered human IgG1 monoclonal antibody specific for the alpha chain of the IL-2 receptor. A pooled analysis of two randomized, double-blind studies was performed on the efficacy and safety of daclizumab in renal transplantation, given in addition to standard immunosuppression. Patients receiving their first cadaveric renal allograft were randomized to receive 5 doses of daclizumab (n = 267) or placebo (n = 268), starting pre-operatively. Acute rejection at 1 year occurred less frequently with daclizumab (n = 74, 27.7 %) than with placebo (n = 116, 43.3%) (P = 0.0001). Fewer patients treated with daclizumab required anti-lymphocyte therapy for acute rejection (7.9 % vs. 15.3 %; P = 0.005). Mean cumulative doses of corticosteroids were lower with daclizumab (4133 mg) than with placebo (4562 mg). One year graft survival was 91.4 % with daclizumab, compared with 86.6 % on placebo (P = 0.065), with patient survival of 98.5 % and 95.1 % for daclizumab and placebo respectively (P = 0.022). Daclizumab was well tolerated. No increase in infectious episodes or lymphoproliferative disorders was observed with daclizumab. The incidence of cytomegalovirus infections was similar with daclizumab and placebo (15 % vs. 17.5 %). Therapy with daclizumab significantly reduces acute rejection in renal transplantation and improves patient survival without any increase in morbidity. PMID- 10836654 TI - The founding of ESOT--the historian's first report. PMID- 10836655 TI - Heart transplantation for radiation-associated end-stage heart failure. AB - Radiation-induced heart disease is an increasingly recognized late sequela of mediastinal radiation therapy for malignant neoplasms. We report four cases of heart transplantation for end-stage heart failure induced by mediastinal radiation therapy. Short-term and intermediate-term results are excellent with all four patients currently surviving a mean of 48 months after transplantation. Neither a second malignancy nor recurrence of the primary malignancy has been observed to date. The early results of heart transplantation for end-stage, radiation-induced heart disease are encouraging. PMID- 10836656 TI - Double recurrence of FSGS after two renal transplants with complete regression after plasmapheresis and ACE inhibitors. AB - A patient who had undergone a first cadaveric donor kidney transplantation for idiopathic focal segmental glomerular sclerosis (FSGS), had an immediate recurrence of a biopsy-proven FSGS that eventually led to graft failure within 5 years from transplantation. The patient underwent a second cadaveric transplantation 10 months later. An immediate recurrence of a biopsy-proven FSGS occurred that was treated with two protracted cycles of plasmapheresis of seven months each, with the addition of an ACE inhibitor from the beginning. A complete and stable remission of FSGS was observed, which continues after more than 6 years from the end of plasmapheresis. The recurrence of FSGS after a second transplantation has a poor prognosis, but prolonged plasmapheresis treatment, by removing circulating factors altering glomerular permselectivity, and the addition of ACE inhibitors, through their potential interference with TGF-beta, might be synergistic in obtaining permanent remission. PMID- 10836657 TI - Daclizumab as escape therapy for late acute kidney rejection in the presence of FK506 nephrotoxicity. PMID- 10836658 TI - Third- and second-generation oral contraceptives are associated with similar risk estimates for venous thromboembolism. AB - A scientific discussion between 1995 and 1999 addressed the question whether second- and third-generation oral contraceptives (OCs) were associated with different risks of venous thromboembolism (VTE). Results from three epidemiological studies became available in the course of 1995, in which such differences were observed. Although it was unclear at that time whether these observations reflected causality or were induced by bias and/or confounding, some regulatory bodies in Europe restricted the indication for use of third-generation oral contraceptives. Immediate media attention generated a pill scare in those, but also other, countries. Indications for the influence of bias were observed in the initial studies of 1995 and further substantiated in subsequent utilization and prescribing surveys. The most important bias seemed to be related to differences in age and duration of use between third- and second-generation OC users. A number of new studies as well as new analyses in two of the 1995 databases included measures to limit the effect of the identified biases/confounders. These studies observed similar risks of venous thromboembolism with second- and third-generation oral contraceptives. Two other recently published studies did not or could not include the same level of control for confounding and reported similar results as the 1995 studies, thus reconfirming the relevance of the identified confounders. Population data show that the massive switch in the UK from third-generation OCs to second-generation OCs in 1995 has not resulted in a reduction of the incidence of VTE in OC users after 1995, illustrating that the risk of VTE is not determined by the type of low-dose pill used. In addition, data from trend analyses, spontaneous reporting and studies addressing hemostatic mechanisms in pill-users also do not support a potential difference in risk of VTE between users of second- and third-generation OCs. PMID- 10836659 TI - Inhibition of ovulation by a novel progestogen (drospirenone) alone or in combination with ethinylestradiol. AB - OBJECTIVE: To investigate ovulation inhibition with drospirenone, a novel progestogen that has a profile similar to natural progesterone, when given alone or in combination with ethinylestradiol. METHOD: Hormonal parameters (LH, FSH, 17beta-estradiol and progesterone) and peripheral parameters (cervical score, spinnbarkeit and crystallization), as well as follicle size assessed by ultrasonography, were measured in two groups of healthy women. Forty-eight women aged 19-35 years were randomly assigned to receive 0.5 mg, 1.0 mg, 2.0 mg or 3.0 mg of drospirenone over a single treatment cycle, and 52 women aged 20-35 years were randomized to receive either 2 mg drospirenone/30 microg ethinylestradiol or 3 mg drospirenone/30 microg ethinylestradiol over three treatment cycles. Baseline measurements were taken during a control pretreatment cycle. RESULTS: Adequate ovarian suppression with drospirenone alone was evident at dose levels of 2 and 3 mg, and at 3 mg all subjects had anovulatory cycles. Although both combined preparations (2 mg and 3 mg drospirenone/30 microg ethinylestradiol) inhibited the hypothalamic-pituitary-ovarian axis, follicular maturation leading to escape ovulation was observed in three subjects in the 2 mg drospirenone/30 microg ethinylestradiol group. Only one of these ovulations was considered to be definitely the result of treatment failure. All cycles in the 3 mg drospirenone/30 microg ethinylestradiol group were anovulatory. No statistically significant difference was found between treatment groups. CONCLUSION: The combination of 3 mg drospirenone/30 microg ethinylestradiol (Yasmin, Schering AG) reliably inhibits ovulation, with a low frequency of follicular maturation, and provides a reasonable safety margin. PMID- 10836660 TI - Efficacy and tolerability of a monophasic oral contraceptive containing ethinylestradiol and drospirenone. AB - OBJECTIVE: To assess the contraceptive reliability, cycle control and tolerability of a new monophasic oral contraceptive containing 30 g ethinylestradiol plus 3 mg drospirenone (Yasmin, Schering AG, Berlin, Germany), it was compared with an established oral contraceptive containing 30 g ethinylestradiol plus 150 g desogestrel (Marvelon, NV Organon, Oss, The Netherlands). METHODS: A randomized, open-label, 13-cycle study was performed at 80 European centers. Contraceptive reliability, cycle control, blood pressure, body weight, the incidence of adverse events and skin condition were assessed during 13 cycles of oral contraceptive use, and at follow-up. Subjects recorded body weight on three consecutive days pretreatment and weekly thereafter. RESULTS: Of 2069 women who started the study and received the trial preparations in a ratio of 4:1 (ethinylestradiol/drospirenone, n = 1657; ethinylestradiol/desogestrel, n = 412), 1615 completed the 13 cycles plus follow up, providing data for over 23,000 evaluable cycles. Eleven pregnancies occurred during treatment, only one of which (in the ethinylestradiol/drospirenone group) could not be ascribed to user failure or interaction with other factors. Both preparations provided effective contraception and cycle control. Pre-existing acne and seborrhea were improved and blood pressure was essentially unchanged. The two treatments differed in their effect on body weight, the difference being statistically significant. In the ethinylestradiol/drospirenone group, there was a distinct decrease over the whole treatment phase, while a subtle and less distinct decrease was documented in the ethinylestradiol/desogestrel group. CONCLUSIONS: The combination of 30 g ethinylestradiol/3 mg drospirenone provides effective oral contraception, excellent cycle control, good tolerability and a level of weight loss that may have a significant beneficial effect on compliance in women with a tendency to weight gain due to water retention. PMID- 10836661 TI - Multicenter study of the efficacy, cycle control and tolerability of a phasic desogestrel-containing oral contraceptive. AB - OBJECTIVE: To investigate the efficacy, cycle control and tolerability of a phasic oral contraceptive containing ethinylestradiol 35/30/30 microg and desogestrel 50/100/150 microg. METHODS: A multicenter study was conducted involving 2070 healthy, fertile women, who received study treatment for six treatment cycles. RESULTS: Most of the participants (79%) had previously been using an alternative oral contraceptive. In 10,408 treatment cycles, two women became pregnant while on treatment (Pearl index, 0.25). The incidence of irregular bleeding was 10% before treatment, rising to 27% at cycle 1, and decreasing to 11% by cycle 6. Irregular bleeding was mainly due to spotting rather than breakthrough bleeding and the incidence of breakthrough bleeding remained below 2.2% for most of the study period. Only 1.8% of women withdrew due to bleeding irregularities. First-time oral contraceptive users initially experienced more irregular bleeding than switchers but these differences lessened over time. The most common adverse events during treatment were headache, breast tenderness and nausea. The incidence of these adverse events fell to below pretreatment levels with continued use. CONCLUSION: The phasic preparation was effective and well tolerated. PMID- 10836662 TI - Vaginal misoprostol 600 microg for early abortion. AB - The objective of this study was to evaluate the efficacy and safety of the vaginal self-administration of 600 microg misoprostol up to a maximum administration of three doses in a 24-h period, one every 8 h, for abortion up to 9 weeks' gestation. A group of 90 voluntary women with gestations from 35 to 63 days participated in the study. All women who aborted received a single additional dose of 600 microg misoprostol. Outcome measures included successful abortion (complete abortion without requiring surgical procedure) and side effects. Complete abortion occurred in 57/89 (64%, 95% confidence interval 53 74%) subjects. The mean expulsion time was 7.4 +/- 3.8 h (median 7.2 h, range 3 20 h) for all women who aborted within the first 24 h of the administration of misoprostol. Thirty-two cases failed to abort, 28 cases due to failure of the method, of which 24 had a negative cardiac rhythm after the third dose, and four cases due to the doctor's decision. The results obtained in this study led us to the conclusion that the 600-microg misoprostol dose regimen was not adequate to produce a high or an acceptable abortive efficacy. PMID- 10836663 TI - Analysis of the underestimation of induced abortions in a survey of the general population in France. AB - INTRODUCTION: A pilot study has been conducted in France to estimate the extent of underreporting of induced abortion and the factors linked to underreporting. METHODS: A representative random sample of 300 women aged between 18 and 44 years of age who had been pregnant during the previous 2 years and 100 women who had not was selected from the telephone directory. Interviews were conducted by telephone. RESULTS: The annual incidence of induced abortion for the 18-44-year old age group was 6.9 per 1000 (95% confidence interval 2.2-11.6), while the rate derived from national statistics was 15 per 1000. Five women who reported a therapeutic abortion in their lifetime had confused induced abortion and therapeutic abortion. There was no significant difference in the proportion of women reporting induced abortion in their lifetime between those answering with another adult present and those who were alone when answering (13.1% versus 11.1%, p = 0.8). There was also no significant difference according to the sex of the interviewer (10.0% for men versus 12.2% for women, p = 0.6). CONCLUSION: The results suggest that the context in which the questions were asked does not affect the validity of the answers and that there is a problem in the comprehension of the terminology used. Extensive rewriting and reformulation of the questions is required to minimize the underreporting of induced abortion. PMID- 10836664 TI - Local spermicidal contraception: a comparative study of the acceptability and safety of a new pharmaceutical formulation of benzalkonium chloride, the vaginal capsule, with a reference formulation, the pessary. AB - OBJECTIVE: To evaluate comparatively the acceptability and safety of a new pharmaceutical form of benzalkonium chloride, the vaginal capsule, with the pessary form. METHODS: Eighty-nine women were randomized to receive either a benzalkonium chloride vaginal capsule or a benzalkonium chloride pessary prior to sexual intercourse according to an open cross-over design over a 2-month study period. RESULTS: The discomfort caused by delayed leakage or discharge was mild. The mean scores for subjective signs (burning, itching, vulvar pruritus) were also mild and were comparable for the two formulations. The capsule appeared to be slightly superior to the pessary regarding discomfort caused by immediate discharge or leakage, ease of use and acceptance by the woman's partner. Local safety was generally good. Adverse events consisted chiefly of vulvar pruritus and a vaginal or vulvar burning sensation, with a higher incidence being associated with the pessary than the capsule. All symptoms regressed spontaneously after the end of treatment. CONCLUSIONS: This study demonstrated good acceptability and good local safety for the benzalkonium chloride vaginal capsule. It is essential to take these parameters into consideration in the evaluation of any local spermicidal contraceptive, since they play a large part in compliance with, and thus in the efficacy of, the product under assessment. PMID- 10836665 TI - Women's acceptance of an injectable progestin-only contraceptive in a free-choice environment in Turkey. AB - A progestin-only injectable contraceptive has been integrated into the method mix at a well-woman clinic in Turkey. During the period from 1st December 1996 to 1st December 1997, among 3545 new method acceptors at the clinic, 254 women decided to use the injectable, giving an initial acceptance rate of 7%. Continuation rates, however, were quite low: 43.7% at 6 months and only 18.5% at 12 months. Though all the side-effects were mentioned during the counselling sessions, side effects such as amenorrhea and spotting might have been underestimated by the potential users of this method, which was new in the country. A more detailed counselling with a strong emphasis on frequent side-effects might lower the initial acceptance rate, but can help to increase the continuation rates. PMID- 10836666 TI - Sexual behavior of secondary-school students in Slovenia. AB - OBJECTIVES: The aim of this study was to analyze the sexual behavior of secondary school students in Slovenia. METHODS: The research was carried out on a representative sample of 4706 secondary-school students aged 15-19 years in Slovenia. The data were obtained by a self-administered questionnaire in April 1996. RESULTS: The average age of the students was 17.5 years. Most students had experiences in kissing (70%) and caressing (59%); a lower number had experiences in petting (43%). Sexual intercourse had been experienced by 38% of the students. The median age at the first sexual intercourse was 18.5 years. The main motives for the first sexual intercourse were love (45%), accident (22%) and curiosity (15%). Contraceptive methods currently used were condoms (60%), the pill (14%), coitus interruptus (4%), other methods (3%) and no method (19%). According to the students, the most appropriate sources of information on sexuality were friends (26%), parents (19%), different sources (19%) and professionals (15%). CONCLUSION: By the age of 18.5 years, approximately one-half of secondary-school students in Slovenia experience sexual intercourse. Most students currently use effective contraception, condoms being the most popular method. The students expect to receive information about sexuality from friends, parents and professionals, but not from the school environment. PMID- 10836667 TI - Teenage pregnancy and ethnicity in The Netherlands: frequency and obstetric outcome. AB - OBJECTIVES: To study differences in frequency and obstetric outcome of teenage pregnancy (not ending in induced abortion) between the main ethnic groups in The Netherlands. DESIGN: A retrospective cohort study based on the 1990-93 birth cohort in the National Obstetric Registry. SUBJECTS: A total of 10,583 teenagers and 54,501 20-24-year-old women who had a singleton pregnancy and were primiparous. MAIN OBSTETRIC OUTCOME MEASURES: These were perinatal death occurring between the 16th week of pregnancy and 24 h after birth, preterm birth and operative delivery (vaginal extraction and Cesarean section). METHOD: Comparison of the frequency of teenage pregnancy between ethnic groups and by bivariate and multivariate analysis of the three outcome measures between the teenage groups, the teenage groups and ethnically related 20-24-year-old women, and the teenage groups and Dutch 22-24-year-old women. RESULTS: A total of 55.2% of pregnant teenagers had non-Dutch ethnicity compared to 13.8% of all pregnant women. Islamic-Mediterranean teenagers constituted the largest group, one in four of all primiparous Mediterranean women being younger than 20 years of age, followed by black teenagers. Except for Hindustani teenagers, perinatal death occurred in all non-Dutch teenage groups more frequently than in Dutch teenagers, but the differences were only significant for black teenagers (odds ratios of black compared to Dutch teenagers were 2.89 (95% confidence interval (CI) 1.89 4.4) and 1.53 (95% CI 1.19-1.98), respectively). Rates for preterm birth were higher in black and Asian than in Dutch teenagers, but the difference was only significant for black teenagers (odds ratio 1.53, 95% CI 1.19-1.98). Compared to ethnically related 20-24-year-old women, rates of perinatal death and preterm birth were significantly higher in Dutch, black and Asian teenagers and, for preterm birth only, in Mediterranean teenagers. Correction for preterm birth showed that only part of these differences in perinatal death could be explained by preterm birth. Vaginal extraction and Cesarean section occurred less frequently in teenagers than in ethnically related (and in Dutch) 20-24-year-old women. Mediterranean teenagers had the lowest Cesarean section rate and Blacks the lowest vaginal extraction rate. CONCLUSION: Teenage pregnancy in The Netherlands is much more common in minority ethnic groups than in the indigenous population, particularly among Islamic-Mediterraneans and Blacks. Obstetric outcomes vary considerably, these being best in Hindustani and poorest in black teenagers, and being worse in teenagers than in 20-24-year-old women. However, teenagers less often had assisted delivery. PMID- 10836668 TI - Pill knowledge amongst oral contraceptive users in family planning clinics in Scotland: facts, myths and fantasies. AB - OBJECTIVES: To document the extent of understanding about the combined pill amongst oral contraceptive users in Scotland. METHOD: A questionnaire was distributed to 2700 pill users in Scottish family planning clinics prior to their consultation with a clinician for a repeat prescription. RESULTS: Knowledge was patchy and usually incomplete in any given area. CONCLUSION: Counselling and teaching about oral contraceptives are important parts of the clinical consultation; even if the user appears knowledgeable about their contraceptive method, there are likely to be some areas where their knowledge could be improved. PMID- 10836669 TI - Changing screening strategies for genital chlamydia in family planning clinics: a good public health strategy? AB - OBJECTIVE: To estimate the prevalence and factors associated with genital chlamydia in 1996 and to evaluate the introduction of restricted screening criteria based on diagnostic testing and prevention of iatrogenic pelvic inflammatory disease in 1997. PATIENTS: Women attending Liverpool family planning clinics. METHOD: Clinical and demographic data were recorded at routine clinic consultations. Data were analyzed using logistic regression. RESULTS: In 1996, a 5.7% (123/2165) chlamydial prevalence was detected. A significantly lower prevalence was observed in the over-25-year-old age group compared to those aged 16-19 years old (p < 0.0001), and in those with an interval of over 11 months since the last new sexual partner compared to those with a new partner in the past 1-3 months (p < 0.05). A significantly higher prevalence was detected in those only presenting with signs of infection alone compared to those presenting with either symptoms or both signs and symptoms of infection (p < 0.05). Those requesting a termination of pregnancy were at higher risk of genital chlamydia than those attending for contraceptive advice (odds ratio 1.97; 95% confidence interval 1.14-3.39). Using the restricted screening criteria, prevalences of 5.4% (78/1438) and 3.7% (60/1641) were found for 1996 and 1997, respectively. Prevalences were significantly higher in 1996 than 1997 when adjusted for the other factors (p < 0.05). CONCLUSION: The study raised awareness of genital chlamydia among women attending family planning clinics and showed that primary care could make a significant contribution to the prevention of chlamydial infection. It is only by initiating intervention strategies in such settings that genital chlamydia will be controlled effectively in the UK. PMID- 10836670 TI - Immediate postabortal insertion of the frameless IUD: review of current experience. AB - Experience with the frameless Gynefix intrauterine device (IUD) for immediate postabortal insertion is growing. Reports confirm that the design characteristics of the frameless IUD (fixed, frameless and flexible) are responsible for the low expulsion, high effectiveness and high continuation rates. Insertion is easy and safe in the hands of trained providers and appears to be as reliable and effective as when inserted at interval. The results suggest that the frameless IUD may be an important new and affordable option to prevent repeat abortion. PMID- 10836671 TI - Psychopathology in children with intellectual disability. AB - Recent advances are reviewed in understanding the heightened prevalence of psychopathology and maladaptive behavior among children with intellectual disability. Researchers have traditionally emphasized measurement and prevalence issues, using either psychiatric assessments or rating scales to identify the prevalence of various problems in children with intellectual disability. Yet the time is ripe to shift directions, and identify more precisely why children are at increased risk for psychopathology to begin with. Although several "biopsycho social" hypotheses are reviewed, a particularly promising line of work links psychopathology to genetic intellectual disability syndromes. Psychiatric vulnerabilities in several syndromes are reviewed, as are the advantages of phenotypic work for understanding psychopathology among children with intellectual disability more generally. PMID- 10836672 TI - Treatment of obsessive-compulsive disorder in children and adolescents. AB - This paper reviews the treatment of obsessive-compulsive disorder (OCD) in children and adolescents. Focusing on clinical features of the disorder and its treatment particular to pediatric onset, diagnosis, assessment, and behavioral, pharmacological, as well as new investigative treatments are covered. Adaptation of cognitive-behavioral therapy for children and adolescents, use of augmenting agents in drug treatment, and subtyping of OCD cases are developments relevant for current practice. PMID- 10836673 TI - Sex differences, assortative mating, and cultural transmission effects on adolescent delinquency: a twin family study. AB - The twin study literature contains inconsistencies regarding both the estimates of genetic, shared, and nonshared environmental influence on delinquency and sex differences in the etiological influences on delinquency. Additionally, little is known about assortative mating and cultural transmission effects on delinquency. The present study was aimed at examining the etiology of delinquency in adolescence and examining assortative mating and transmission effects. To this end, we applied the social homogamy structural equation model to self-reported delinquency data from 486 families with 17-year-old male and female twins. We expected stronger shared environmental influences on delinquency among girls than boys and stronger environmental than genetic effects on adolescent delinquency in general. The results indicated nonsignificant sex differences in the estimates of genetic and environmental influences on delinquency. The best-fitting model suggested that 18 %, 56 %, and 26% of the variance in delinquency among both boys and girls is associated with additive genetic, nonshared environmental, and shared environmental factors, respectively. This finding was consistent with our expectation that environmental factors play an important role in adolescent delinquency. Parental cultural effects accounted for a small part of the transmission of delinquency within families while assortative mating was, in part, accounted for by cultural and social background factors. Future work should focus on identifying important environmental factors related to delinquency. PMID- 10836674 TI - Twenty years' research on peer victimization and psychosocial maladjustment: a meta-analytic review of cross-sectional studies. AB - Cross-sectional quantitative designs are often used to investigate whether peer victimization is positively related to psychosocial maladjustment. This paper presents a meta-analytic review of cross-sectional studies, published between 1978 and 1997, of the association of peer victimization with psychosocial maladjustment. Mean effect sizes were calculated for the association between peer victimization and each form of maladjustment (depression, loneliness, generalized and social anxiety, and global and social self-worth) assessed. The results suggested that victimization is most strongly related to depression, and least strongly related to anxiety. There was no evidence that victimization is more strongly related to social than to psychological forms of maladjustment. Effect sizes were stronger when the same informants were used to assess both victimization and maladjustment than when different informants were used. There were some design limitations to the studies reviewed, but all together their results provide a strong background for more complex research into the course and treatment of victims' distress. PMID- 10836675 TI - Applications of the expressed emotion construct to young children with externalizing behavior: stability and prediction over time. AB - We explored the utility of the Expressed Emotion (EE) construct with a community sample of young children (N = 91), studying the stability of EE scores over a 2 year period and the extent to which EE was associated with simultaneous and subsequent disruptive behavior. As part of a longitudinal study, families were assessed when the child was in preschool, 1st grade, and 3rd grade. Maternal EE was measured at preschool and 1st grade using the Five Minute Speech Sample and behavior ratings were obtained using the Child Behavior Checklist. At 3rd grade, the Diagnostic Interview Schedule for Children was used to determine DSM-IV diagnoses. The stability of the EE rating over the 2-year period from preschool to 1st grade was statistically significant although modest. At 1st grade, EE ratings were significantly related to the extent of externalizing behavior problems. Regression analyses that controlled for maternal stress levels determined that preschool EE ratings predicted classification of ADHD over 4 years later, at 3rd grade. The relationships between EE and child problem behavior were almost exclusively determined by the EE criticism dimension; the emotional overinvolvement dimension was not related to child behavior. The implications of these findings are discussed. PMID- 10836676 TI - Psychopathology and short-term emotion: the balance of affects. AB - In this study, the relationship between short-term emotion expressions and dimensional ratings of internalizing and externalizing symptomatology was examined. Short-term emotions, defined as facial or vocal displays of emotion generally lasting less than 10 seconds and elicited by a specific and proximal event, were observed during recess in 71 children from diverse socioeconomic backgrounds, who were between 4 and 8 years old. Internalizing and externalizing symptomatology was assessed through parent and teacher questionnaire. Sociometric ratings were obtained from peers on children's anger and aggression. It was hypothesized following Tomkins (1979) and others that one affect becomes predominant in the emotional experience of the individual. Different operationalizations of this concept were examined. Using regression analyses, externalizing symptomatology was found to be predicted by higher levels of anger, lower levels of happiness, and lower levels of sadness. Internalizing symptomatology was found to be predicted by higher levels of sadness and lower levels of anger. It was concluded on the basis of these data that the relationship between short-term emotion and internalizing and externalizing psychopathology is best understood as the balance between different short-term emotions. Results are discussed in the context of theories of emotion and their functions. PMID- 10836677 TI - The association of reading disability, behavioral disorders, and language impairment among second-grade children. AB - Children with language impairment (LI) have been shown to be at risk for reading disability (RD) and behavior disorder (BD). Previous research has not determined the specific pattern of these conditions associated with LI. This study sought to determine if the behavior disorder and reading problems represented different outcomes or if these conditions occurred together when found with LI. A group of 581 second-grade children, including 164 children with LI, were examined for spoken language, reading, and behavior disorder. The data for each of these areas were examined as dimensional traits and as clinical categorical traits. Reading and spoken language were found to be strongly correlated (r = .68); RD was found in 52 % of the children with LI and in only 9 % of the controls. Scores of parent ratings for BD were also significantly correlated with spoken language scores (r = .29). Clinical levels of BD were found in 29% of the children with LI and 19% of the controls. An examination of the co-occurrence of clinical levels of BD, RD, and LI showed BD in children with LI to be conditioned by the child's reading status. The data indicated that whereas RD was directly associated with BD, the association of LI with BD required the mediation of RD. PMID- 10836678 TI - Good test--retest reliability for standard and advanced false-belief tasks across a wide range of abilities. AB - Although tests of young children's understanding of mind have had a remarkable impact upon developmental and clinical psychological research over the past 20 years, very little is known about their reliability. Indeed, the only existing study of test-retest reliability suggests unacceptably poor results for first order false-belief tasks (Mayes, Klin, Tercyak, Cicchetti, & Cohen, 1996), although this may in part reflect the nonstandard (video-based) procedures adopted by these authors. The present study had four major aims. The first was to re-examine the reliability of false-belief tasks, using more standard (puppet and storybook) procedures. The second was to assess whether the test-retest reliability of false-belief task performance is equivalent for children of contrasting ability levels. The third aim was to explore whether adopting an aggregate approach improves the reliability with which children's early mental state awareness can be measured. The fourth aim was to examine for the first time the test-retest reliability of children's performances on more advanced theory-of mind tasks. Our results suggest that most standard and advanced false-belief tasks do in fact show good test-retest reliability and internal consistency, with very strong test-retest correlations between aggregate scores for children of all levels of ability. PMID- 10836679 TI - Variable expression of the autism broader phenotype: findings from extended pedigrees. AB - Factors influencing the rate, form, and severity of phenotypic expression among relatives of autistic probands are examined. Family history data on 3095 first- and second-degree relatives and cousins from 149 families with a child with autism and 36 families with a child with Down syndrome are studied. The results provide further evidence of an increased risk among autism relatives for the broadly defined autism phenotype. Of proband characteristics, severity of autism and obstetric optimality were confirmed as being related to familial loading for probands with speech. There was little variation in loading among probands lacking speech. The type of phenotypic profile reported in relatives appeared little influenced by characteristics of the relative or the proband, except for variation by degree of relative, parental status of relative, and perhaps proband's birth optimality score. Phenotypic rates among parents suggested reduced fitness for the severest and more communication-related forms of expression but not for the more mild and social forms of expression. Patterns of expression within the families did not support a simple X-linked nor an imprinted X-linked mode of inheritance. The basis for sex differences in rates of expression is discussed. PMID- 10836680 TI - The long-term psychological effects of a disaster experienced in adolescence: I: The incidence and course of PTSD. AB - Previous studies have shown that children and adolescents exposed to traumatic experience in a disaster can suffer from high levels of post-traumatic stress. The present paper is the first a series reporting on the long-term follow-up of a group of young adults who as teenagers had survived a shipping disaster-the sinking of the "Jupiter" in Greek waters-between 5 and 8 years previously. The general methodology of the follow-up study as a whole is described, and the incidence and long-term course of Post-Traumatic Stress Disorder (PTSD). It is the first study of its kind on a relatively large, representative sample of survivors, using a standardised diagnostic interview, and comparing survivors with a community control group. Survivors of the Jupiter disaster (N = 217), and 87 young people as controls, were interviewed using the Clinician Administered PTSD Scale (CAPS). Of the 217 survivors, 111 (51.7%) had developed PTSD at some time during the follow-up period, compared with an incidence in the control group of 3.4 % (N = 87). In the large majority of cases of PTSD in the survivors for whom time of onset was recorded, 90 % (N = 110), onset was not delayed, being within 6 months of the disaster. About a third of those survivors who developed PTSD (30%, N = 111) recovered within a year of onset, through another third (34 %, N = 111) were still suffering from the disorder at the time of follow-up, between 5 and 8 years after the disaster. Issues relating to the generalisability of these findings are discussed. PMID- 10836681 TI - The long-term psychological effects of a disaster experienced in adolescence: II: General psychopathology. AB - Children and adolescents exposed to trauma can suffer major adverse psychological effects including not only post-traumatic stress but also other psychological disorders. This study investigates the long-term course of general psychopathology following trauma in adolescence using a standardised diagnostic interview and comparisons with a matched control group. Young people (N= 216) who as teenagers had survived a shipping disaster-the sinking of the "Jupiter" in Greek waters-between 5 and 8 years previously and 87 young people as matched controls were interviewed. The survivors showed raised rates of diagnosis in a range of anxiety and affective disorders during the follow-up period. The highest rates were among the survivors who had developed Post-Traumatic Stress Disorder (PTSD), and those survivors who had not were generally similar to the controls. Onset of anxiety and affective disorders varied between being indefinitely close to the disaster to years later. Differences in rates of disorder between the survivor and control groups had lessened by the time of follow-up but were still apparent, due to continuing distress among the survivors still suffering from PTSD, and to a lesser extent among those who had recovered from PTSD. Generalisability of the findings are discussed. PMID- 10836682 TI - Visual behaviour of ADHD children during an attention test: an almost forgotten variable. Attention-Deficit Hyperactivity Disorder. AB - The goal of this study was to examine whether looking away behaviour of ADHD children interferes with their test performance. ADHD and normal children carried out two continuous performance tests (CPTs): one with a regular interstimulus interval (ISI), and the other with an irregular ISI. Children were instructed to push a response button when a target stimulus was presented on the monitor. The children's visual behaviour was recorded and scored offline. A micro-analysis of the visual behaviour indicated that ADHD children timed their looking away behaviour in the regular CPT: i.e. they looked away from the monitor and back in the interval between two succeeding stimuli. As a result they did not miss stimuli. Timing of looking away was less possible in the CPT with the irregular ISI. In this condition, looking away interfered with the ADHD children's task accuracy. In sum, looking away behaviour had a negative effect on the accuracy of test performance of ADHD children when stimuli were unpredictable. Looking away behaviour was not associated with the slower reaction times of the ADHD children. Hence, the often reported slowness of ADHD children is not to be explained by their visual behaviour. PMID- 10836683 TI - Quantifying leukocyte and thrombocyte suppression-recovery after Re-186 HEDP for bone metastases. AB - Using a standard quantity of Re-186 HEDP, for therapy of osseous metastases from carcinoma of the breast, Palmedo and associates reported leukocyte and thrombocyte responses at 4 and 8 weeks. These were analyzed by dividing results into categories, based on the numeric initial values. Thrombocytes at 8 weeks exceeded pretreatment values in 10 of 18 cases. This "over-rebound" was also found in 7 of 19 leukocyte responses. The ability to predict thrombocyte leukocyte response, in most cases, was linked to the initial hematologic values. If coupled with data on the status of bone marrow, this may allow the quantity of therapeutic radiopharmaceutical used to be increased or decreased without irreversibly compromising hematopoietic responses. PMID- 10836684 TI - The role of bone scanning in the detection of metastatic calcification: a case report. AB - Metastatic calcification associated with renal failure is well described. Bone scanning agents accumulate to various degrees within extraskeletal sites of metastatic calcification. The authors describe a patient with polycystic kidney disease resulting in renal failure, with the subsequent development of secondary hyperparathyroidism and metastatic calcification. Bone scintigraphy revealed abnormal uptake in both lungs, the right leg, and the right hand. PMID- 10836685 TI - Tc-99m MIBI to evaluate children with Ewing's sarcoma. AB - PURPOSE: Tc-99m MIBI has been used increasingly to evaluate benign and malignant tumors because of its tumor-seeking properties and ability to provide an imaging assessment of multiple-drug resistance. This study investigated the clinical utility of Tc-99m MIBI in the management of Ewing's sarcoma in children. METHODS: Thirteen Tc-99m MIBI studies in nine (six male, three female) patients ages 6.5 to 20 years (mean, 13.4 years) with Ewing's sarcoma were reviewed. All patients had imaging studies at diagnosis, and four had follow-up studies during or after therapy. Scintigraphy was evaluated for Tc-99m MIBI uptake within the tumor and in metastases, which other imaging modalities had shown to be present in four patients. Scintigraphic results were correlated with the clinical course in all patients and with tumor P-glycoprotein status in six patients. RESULTS: Tc-99m MIBI accumulated in 6 of 9 primary tumors and did not accumulate in one recurrent tumor. No metastases showed Tc-99m MIBI uptake. The presence or absence of Tc-99m MIBI uptake at diagnosis or after therapy carried no prognostic significance. Tc 99m MIBI was present in the two tumors that were P-glycoprotein positive and in only one of four tumors that were P-glycoprotein negative. CONCLUSION: Tc-99m MIBI imaging does not appear to be useful in Ewing's sarcoma. PMID- 10836686 TI - The role of bone scintigraphy in patients with Erdheim-Chester disease. AB - Erdheim-Chester disease (ECD) is a rare disorder that has been reported fewer than 60 times in the literature. Although clinical findings seem to be specific at first sight, histologic classification remains unclear. It has not been decided whether ECD is part of the spectrum of histiocytoses or whether it may be a lipid storage disorder or even a primary macrophage cell disorder, although it does show a distinct histologic pattern. However, the clinical appearance alone shows several typical features, rendering the diagnosis very probable if present. This article illustrates the importance of bone scanning in ECD, because the scintigraphic pattern of involved skeletal sites may in themselves lead to the diagnosis. Several differential diagnoses are considered. The importance of bone scintigraphy as an imaging method in patients with an unclear diagnosis is discussed, as exemplary in ECD, as is its role for the detection of sites of skeletal involvement in other diseases. PMID- 10836687 TI - Unusual case of diagnosis of pigeon breeder's hypersensitivity pneumonitis with ventilation-perfusion lung scan. AB - The authors describe a patient thought to have pulmonary embolism. Results of a perfusion scan were normal, but Tc-99m DTPA aerosol ventilation lung scan showed, in successive views, decreased activity of the deposited aerosol. The lung Tc-99m DTPA clearance was measured and was markedly increased. This suggested possible hypersensitivity pneumonitis, which was later confirmed as pigeon breeder's disease. The decreased activity observed on the DTPA ventilation scan must be directed toward increased lung clearance and its measurement. PMID- 10836688 TI - Acute axillosubclavian vein thrombosis (Paget-Schroetter syndrome) detected by Tc 99m MAA during pulmonary perfusion scintigraphy. AB - The authors describe a 24-year-old man who reported the sudden onset of dyspnea and swelling of his left upper arm. An area of increased activity in the left axillosubclavian region evident on pulmonary perfusion scintigraphy with Tc-99m MAA suggested Paget-Schroetter syndrome (primary deep venous thrombosis of the upper extremity). PMID- 10836689 TI - Life-threatening hemorrhage of an unanticipated superficial circumflex iliac artery origin imaged with Tc-99m-labeled erythrocytes. AB - The authors report, for the first time, the nuclear medicine diagnosis of extraperitoneal bleeding that originated from the superficial circumflex iliac artery and review the critical role of dynamic nuclear imaging techniques in the localization of sites of nonenteric hemorrhage. Dynamic imaging techniques using Tc-99m-labeled erythrocytes played a critical role in the localization of an extraperitoneal bleeding site near the entrance site of a Jackson-Pratt drain. Localization of the site remained undiagnosed before nuclear imaging despite two exploratory laparotomies and diagnostic angiography. PMID- 10836690 TI - Contralateral cortical diaschisis in a patient with cerebellar astrocytoma after radiation therapy. AB - Contralateral cerebellar diaschisis, hypometabolism in the cerebellum contralateral to a cortical lesion, is a well described phenomenon in patients with stroke and brain tumor. However, few reports exist of patients with cerebellar lesions with the finding of contralateral cortical hypometabolism. The authors present a case of a patient with a right cerebellar astrocytoma after surgical resection and radiation therapy in which fluorine-18 fluorodeoxyglucose positron emission tomographic imaging, performed to rule out recurrent tumor, revealed global hypometabolism in the left cerebral cortex. The concept of contralateral cortical diaschisis and possible mechanisms and clinical implications are reviewed. PMID- 10836691 TI - Scintimammographic findings of in situ ductal breast carcinoma in a double-phase study with Tc-99m(V) DMSA and Tc-99m MIBI value of Tc-99m(V) DMSA. AB - The authors present a case of in situ ductal carcinoma of the breast (DCIS) with no associated mass in a 46-year-old woman examined with Tc-99m MIBI and Tc-99m(V) DMSA scans, which were acquired in separate sessions 10 minutes and 60 minutes after injection. Histologic analysis revealed a small (<1 cm) infiltrating ductal carcinoma located within the DCIS. Mammography showed a cluster of microcalcifications on a very dense parenchymal background. Tc-99m(V) DMSA was characterized as positive for DCIS, especially in the delayed image. Tc-99m MIBI failed to identify the lesions previously noted. In conclusion, Tc-99m(V) DMSA scintimammography seems to have an advantage and could improve the detection of nonpalpable in situ breast carcinomas. PMID- 10836692 TI - Use of fusion images of I-131 metaiodobenzylguanidine, SPECT, and magnetic resonance studies to identify a malignant pheochromocytoma. AB - Pheochromocytoma is a chromaffin tumor in which 10% are extra-adrenal and 10% are malignant. I-131 metaiodobenzylguanidine (MIBG) scintigraphy has an important role in the identification of these tumors and investigation of metastatic lesions. The authors describe a 36-year-old woman who underwent resection of a malignant left adrenal pheochromocytoma who was thought to have metastases in the liver and para-aortic lymph nodes. Fusion images of I-131 MIBG SPECT and magnetic resonance studies were obtained to properly identify the metastatic lesions. These fusion images helped greatly in subsequent surgery. PMID- 10836693 TI - Whole-body Tc-99m sestamibi scintigraphy in the follow-up of differentiated thyroid carcinoma. AB - PURPOSE: This study evaluated the potential of Tc-99m sestamibi whole-body scan (WBMIBI) as an alternative to whole-body I-131 scan (WBI) for the follow-up of patients with differentiated thyroid carcinoma. MATERIALS AND METHODS: We evaluated 99 consecutive patients with differentiated thyroid carcinoma who had total or nearly total thyroidectomy followed by an ablative dose of I-131 (86 women, 13 men; mean age, 44 +/- 12 years). WBMIBI was performed and serum thyroglobulin (TG) levels were obtained at least 6 months after I-131 treatment. All persons were receiving levothyroxine therapy. RESULTS: From the total of 110 studies performed, WBMIBI and TG were in agreement in 96% and discordant in 4%. From the 27 crossed studies (WBMIBI x TG) with at least one abnormal result, 16 were compared with WBI. In four cases, the WBI did not reveal functioning thyroid tissue when both TG and WBMIBI indicated tumoral activity. In one case of pulmonary metastasis confirmed by chest radiographs, with a normal TG value, the results of both WBMIBI and WBI were positive. CONCLUSIONS: WBMIBI should be considered as a scintigraphic method in the follow-up of differentiated thyroid carcinoma. This technique can show the sites of tumoral activity with optimal image resolution, particularly in those with abnormal TG and negative WBI results, and it is a potentially valuable tool in patients with anti-TG antibodies. The WBI in patients having ablation should be reserved only for therapy planning. PMID- 10836694 TI - Use of indium-111-labeled hepatocytes to determine the biodistribution of transplanted hepatocytes through portal vein infusion. AB - PURPOSE: Hepatocyte transplantation is useful for ex vivo gene therapy and liver repopulation. Methods for hepatic reconstitution were recently developed, but hepatocyte transplantation systems must be optimized. The authors report their experience with In-111 oxyquinolone labeling of a test dose of hepatocytes (108 cells) for noninvasive assessment of the biodistribution of transplanted hepatocytes in a 5-year-old child with omithine transcarbamoylase deficiency. MATERIALS AND METHODS: Donor hepatocytes (approximately 108) were radiolabeled using a commercially available In-111 oxyquinolone solution (specific activity of 1 mCi/ml). RESULTS: The overall labeling efficiency was 36.4%. A final dose of approximately 290 ,uCi of the In-111-labeled hepatocytes in 10 ml serum-free phosphate-buffered saline was infused percutaneously into the portal vein approximately 2.5 hours after their preparation. The study was performed 3 hours before cell transplantation (109 cells). Quantitative analysis of the biodistribution of In-111-labeled hepatocytes indicated that cells were predominantly localized in the liver immediately after portal vein-infused transplantation. The predominant hepatic distribution was persistent for as long as 7 days after the procedure, with an average liver-to-spleen ratio of 9.5 to 1. No significant pulmonary radiotracer uptake was present. CONCLUSION: These results indicate that In-111 labeling of hepatocytes is useful for the short-term noninvasive analysis of the biodistribution of transplanted hepatocytes. PMID- 10836695 TI - Radionuclide lymphangioscintigraphy in the evaluation of peripheral lymphedema. AB - PURPOSE: The primary difficulty in evaluating and treating peripheral lymphedema is visualization of the lymphatics. Functional lymphatic studies have been performed on patients with peripheral edema to diagnose lymphedema, to determine its severity, and to understand the varied drainage patterns. METHODS: After intradermal injection in the hands or feet, initial flow and whole-body images were taken using Tc-99m human serum albumin in more than 700 patients with possible lymphedema. RESULTS: Clear images of truncal lymph transport and draining lymph nodes were obtained, and pattern differences between primary and secondary lymphedema were seen. Follow-up studies showed any functional change in lymphatic dynamics. CONCLUSION: Peripheral lymphatics can now be easily visualized. Because lymphangioscintigraphy can be performed before and after medical treatment, follow-up evaluation of patients with lymphedema is possible. The procedure is noninvasive, repeatable, easy to perform, and harmless to the lymphatic endothelium. PMID- 10836696 TI - Functioning liver metastases on an I-131 whole-body scan: a case of malignant struma ovarii. AB - A 46-year-old women was examined for severe constipation. Pelvic examination revealed a large pelvic mass extending to the level of the umbilicus. Computed tomography showed a large multicystic, septated mass in the pelvis and a small amount of fluid in the cul de sac. In addition, multiple ill-defined, mixed attenuation hepatic lesions were identified. A malignant ovarian neoplasm with liver metastases was considered, so the pelvic mass was resected. Interestingly, histopathologic analysis revealed malignant struma ovarii of the follicular type. Biopsy of the liver lesions confirmed metastatic disease with similar histopathologic findings. All thyroid laboratory values were in the normal range. The patient then had a total thyroidectomy to optimize thyroid ablation therapy with I-131. This revealed a small follicular adenoma but no evidence of cancer. An I-131 whole-body scan was performed and showed uptake in multiple functioning liver metastases. PMID- 10836697 TI - Tc-99m pertechnetate scintigraphy before and after potassium perchlorate administration for the diagnosis of retrosternal goiter. AB - A 52-year-old woman was hospitalized because of dyspnea and dysphagia. Thoracic computed tomography revealed a retrotracheal mass. Tc-99m pertechnetate scintigraphy showed intense accumulation of radioactivity corresponding to the mediastinal mass detected by computed tomography. Repeated Tc-99m pertechnetate scintigraphy performed after oral administration of potassium perchlorate (KCLO4) revealed complete disappearance of the radioactive accumulation in the mediastinum, suggesting that the retrotracheal mass was a retrosternal goiter. Subsequent surgical removal and analysis of the mass showed it was indeed a retrosternal goiter. This case highlights the importance of Tc-99m pertechnetate thyroid scintigraphy with and without KCLO4 administration as a simple, accurate, and cost-effective imaging method to diagnose retrosternal goiter. PMID- 10836698 TI - Incidental detection of a malignant hurthle cell carcinoma by Tc-99m sestamibi cardiac imaging. AB - A 60-year-old man reported chest pain and shortness of breath. His medical history was negative for myocardial infarction but positive for "mini strokes" and type 2 diabetes mellitus. Tc-99m sestamibi cardiac imaging revealed an abnormal focus of increased activity in the left lobe of the thyroid. Although no cardiac abnormalities were found, a I-123 thyroid scan subsequently showed a solitary hypofunctioning nodule involving the middle and inferior aspects of the left lobe of the thyroid, which fine-needle aspiration proved was a Hurthle cell carcinoma. PMID- 10836699 TI - Tc-99m MAG3 scintigraphy in unusually dilated bilateral extrarenal pelves. AB - The authors report the findings of a radionuclide diuretic renogram in a patient with markedly dilated pelves bilaterally that filled most of the abdominal cavity. With the patient in the sitting position, at the end of the furosemide injection, there was good delineation and filling of the dilated calyces. These findings are unusual because of the extensive pelvicaliceal dilatation, and they emphasize the importance of the sitting position. PMID- 10836700 TI - Nephroptosis: the Tc-99m glucoheptonate scan as a diagnostic method. AB - A 40-year-old man was examined because he was a potential "healthy" renal donor. However, the routine work-up before surgery revealed hypertension, although there was no family history of this condition. The patient was examined to exclude secondary causes of hypertension. Tc-99m glucoheptonate renal imaging showed nephroptosis of the right kidney when the patient was standing, and this may have been the cause of the hypertension. PMID- 10836701 TI - Radioisotope bone scanning in pulmonary alveolar microlithiasis. AB - A 30-year-old woman was examined for a history of exertional breathlessness, swelling of her feet, and a mild dry cough of 4 to 5 months' duration. Her symptoms developed during the last month of her pregnancy, with gradually increasing dyspnea, swelling of the feet, and reduced urinary output. There was no history of expectoration, hemoptysis, chest pain, or tuberculosis. General physical examination showed no evidence of clubbing of the nails or lymphadenopathy. Chest auscultation revealed a few end-inspiratory crepitations at both lung bases. Bronchial alveolar lavage showed calcium particles, whereas results of the transbronchial lung biopsy were consistent with alveolar microlithiasis. PMID- 10836702 TI - Bilateral accessory iliac horns: pathognomonic findings in Nail-patella syndrome. Scintigraphic evidence on bone scan. AB - Nail-patella syndrome is a rare hereditary (autosomal dominant) disorder, also called hereditary osteo-onychodysplasia and Fong's syndrome. Its incidence is 4.5 per million population in the United States. Patients have a characteristic tetrad of pathologic symptoms including fingernail dysplasia, hypoplastic or absent patellas, radial head dislocation, and iliac horns. Soft-tissue changes and renal dysplasia have also been associated with the syndrome. Iliac horns are bilateral accessory outgrowths consisting of cortex and medulla continuous with the iliac bone. They are located at the site of attachment of the gluteus medius muscles and project posterolaterally. These smooth bony outgrowths are asymptomatic, frequently palpable, and, because they have no effect on gait, they need not be treated. Iliac horns are the pathognomonic feature of Nail-patella syndrome; that is, they occur in approximately 80% of cases and are observed only in this condition. Four patients (two female, two male) with Nail-patella syndrome have been examined in the authors' department: three family members, including a 37-year-old woman, her 18-year-old son, and her 15-year-old daughter, and an unrelated 26-year-old man. All patients, regardless of age or sex, had similar pelvic findings on their bone scans. Although whole-body scans were obtained in all patients, significant scintigraphic findings were observed only in the pelvis in all the patients examined. This is most likely the result of the mild nature of the related deformities, which do not yield scintigraphically detectable osteoblastic changes. A representative image showing these independent ossification on a Tc-99m MDP bone scan is presented. A conventional pelvic radiographic image of the same patient's pelvis is presented for comparison. PMID- 10836703 TI - Monostotic fibrous dysplasia of the sphenoid sinus: a serendipitous finding on a bone scan. AB - A 22-year-old woman had a Tc-99m MDP whole-body scan for low back pain. A focal area of increased activity was seen in the skull base in the region of the sella turcica. A computed tomographic examination showed ground-glass opacification of the sphenoid sinus and bony sclerosis along its walls, characteristic of fibrous dysplasia. Monostotic fibrous dysplasia, the more common form compared with the polyostotic variety, occurs in 70% to 80% of all patients with fibrous dysplasia. Monostotic lesions usually involve the ribs, femur, tibia, cranium, maxilla, and mandible. The frontal and sphenoid bones are the cranial bones most commonly involved. PMID- 10836704 TI - Unusual presentation of solitary bone metastasis from breast carcinoma mimicking acute osteomyelitis of the left midtibial shaft. AB - The authors report an unusual presentation of a solitary bone metastasis in the left midtibial shaft with no other skeletal involvement in a patient with breast carcinoma. The incidence of solitary bone metastasis below the knees is rare. Clinically, the lesion was tender when palpated. A bone scan revealed increased blood flow and blood pool activity, with intense midtibial bony uptake in delayed images. These findings are similar to those of acute osteomyelitis. Biopsy revealed bony metastasis from the patient's breast carcinoma. PMID- 10836705 TI - Tc-99m sestamibi and In-111 DTPA octreotide uptake in breast carcinoma with neurendocrine differentiation. AB - Some breast tumors are classified as primary neuroendocrine carcinomas because of argyrophilia and positivity for neuroendocrine markers (chromogranins A and B and neuron-specific enolase), regardless of their cellular rest and cord structures. Tc-99m sestamibi has been widely used to identify epithelial breast carcinoma and lymph node metastases, whereas In-111 DTPA-octreotide has been used to identify primary and secondary neuroendocrine neoplasms specifically. The use of In-111 DTPA-octreotide and Tc-99m sestamibi scintigraphy in a woman with neuroendocrine differentiated cancer of the left breast is reported. Uptake of these radiopharmaceuticals only in the breast tumor permitted identification of a primary breast carcinoma, whereas absence of In-111 DTPA-octreotide uptake in other sites helped to exclude the presence of other neuroendocrine neoplasms in other organs. PMID- 10836706 TI - Tc-99m MDP uptake in uterine leiomyoma. AB - A 47-year-old woman with adenocarcinoma of the right breast had bone scintigraphy with Tc-99m MDP. Bone imaging did not show any metastases. However, a large area of increased tracer uptake was seen extending from the abdomen to the pelvis. Abdominal ultrasound revealed a large solid and heterogeneous mass, measuring 18 x 11 x 14.3 cm, that originated in an empty uterus. A biopsy of the surgical specimen showed a leiomyoma of the uterus. PMID- 10836707 TI - Pyometra as a lower abdominal doughnut sign on a Ga-67 scan. AB - A 77-year-old woman was referred for Ga-67 scan to evaluate intermittent fever and chills that had lasted more than 20 days. The Ga-67 whole-body scan revealed a doughnut-shaped Ga-67 accumulation in the lower abdominal region. Combined Ga 67 and Tc-99m MDP bone scan confirmed that this activity was in the uterus, because the shape of the urinary bladder on bone scan was different from that of the Ga-67-avid lesion. Pyometra was proved during operation, and pus culture was performed. PMID- 10836708 TI - Fluorine-18 fluorodeoxyglucose positron emission tomography correlated with computed tomographic scan and magnetic resonance imaging in a case of hematometrocolpos. AB - A 12-year-old girl had intense abdominal pain that had increased in the past 3 months and was accompanied by weight loss. An ultrasound examination revealed large cystic masses in the abdomen. A computed tomographic scan could not conclusively rule out a malignant condition. The hymen was normal on physical examination, but magnetic resonance imaging confirmed that the abnormalities corresponded to dilated cavities of the vagina, uterus, and fallopian tubes, with an appearance suggestive of hematometrocolpos. Fluorine-18 fluorodeoxyglucose (FDG) positron emission tomography was requested concurrently with the magnetic resonance image to assess the metabolic activity of the lesions and to exclude the presence of distant metastases. Large defects without FDG accumulation were noted in the areas corresponding to the cystic masses. Vaginal atresia with hematometrocolpos was confirmed at surgery. This rare case involving F-18 FDG positron emission tomographic imaging in hematometrocolpos illustrates that this diagnosis should be considered in the presence of symmetric hypometabolic masses in the pelvis. PMID- 10836709 TI - Tc-99m dicarboxypropane diphosphonate uptake in ovarian fibrothecoma. AB - A 76-year-old woman had a palpable mass in the lower abdomen. Computed tomography showed an abnormal pelvic mass with necrotic areas, probably of ovarian origin. A Tc-99m dicarboxypropane diphosphonate whole-body scan performed to evaluate the extent of the disease (i.e., the presence of bone metastases) revealed only diffuse uptake of the tracer in the pelvis. Histopathologic analysis after surgical excision of the mass revealed a fibrothecoma of the right ovary, which is a benign tumor composed of cells derived from the ovarian stroma and, in some instances, resembles the thecal element of the follicle. PMID- 10836710 TI - Discrepancy between Ga-67 citrate and F-18 fluorodeoxyglucose positron emission tomographic scans in pulmonary infection. AB - The authors describe a patient with the acquired immunodeficiency syndrome who had active pulmonary tuberculosis and was receiving anti-tuberculosis treatment. High-grade fever and a right-sided pleural effusion had recently developed. Results of a Ga-67 scan were negative for any focal infection in the chest. Fluorine-18 fluorodeoxyglucose positron emission tomography showed increased uptake in the right lower lung field, which correlated with the diagnosis of concomitant bacterial pneumonia. Anti-tuberculosis treatment can decrease the sensitivity of the Ga-67 scan and could have contributed to this discrepancy. The authors predict that the fluorine-18 fluorodeoxyglucose positron emission tomographic scan will play an important diagnostic role in the management of such a selected group of patients. PMID- 10836711 TI - Afferent loop syndrome: the role of Tc-99m mebrofenin hepatobiliary scintigraphy. AB - Afferent loop syndrome is caused by intermittent mechanical obstruction of the afferent loop of a gastrojejunostomy and may present early as an acute type or late as a chronic type. The authors describe two patients who were examined for a history of bilious vomiting after gastrojejunostomy, and who were thought to have afferent loop syndrome (chronic type) based on clinical findings. Results of routine investigations, such as upper gastrointestinal endoscopy, and ultrasonography were inconclusive. Findings from the barium meal follow-through studies were normal in the first patient and revealed a dilated duodenum in the second patient. Tc-99m bromotriethyl-iminodiacetic acid has been used to identify afferent loop obstruction as represented in these studies. PMID- 10836712 TI - Multiple myeloma showing increased accumulation of Tc-99m hexamethylpropylene amine oxime on brain SPECT. AB - The authors report a case of multiple myeloma with increased accumulation of Tc 99m hexamethylpropylene amine oxime (HMPAO) on brain SPECT. Tc-99m HMPAO is a lipophilic compound that freely passes through the intact blood-brain barrier and cell membrane and is rapidly converted to a hydrophilic form by glutathione and then retained in the neuron for several hours. In general, Tc-99m HMPAO shows decreased accumulation in brain tumors. However, some reports of increased accumulation in brain tumors, such as meningioma, glioblastoma multiforme, high grade astrocytoma, pituitary adenoma, and multiple myeloma, have been published. The Tc-99m HMPAO uptake in these tumors has been attributed to tumor blood flow or glutathione contents within the tumor. With regard to uptake to Tc-99m HMPAO in multiple myeloma, the tumor size is considered to be an additional factor. PMID- 10836713 TI - Current readings in nuclear medicine. PMID- 10836714 TI - Interaction between norepinephrine release and intrarenal angiotensin II formation during renal nerve stimulation in dogs. AB - We examined possible interactions between intrarenal angiotensin II (ANG II) formation and norepinephrine (NE) release during renal sympathetic nerve stimulation (RNS) in anesthetized dogs. During 10 min of continuous RNS (1.5-2 Hz), the ANG II formation rates (ANG II-FR) and NE secretion rates (NE-SR) were determined at 1 and 10 min. Under control conditions, almost the same extent of increase in the NE-SR was observed at 1 and 10 min of RNS, whereas a significant increase in ANG II-FR was observed at 10 min but not at 1 min. During intrarenal arterial infusion of enalaprilat or losartan, the increase in NE-SR and reduction in renal blood flow at 10 min of RNS were suppressed, whereas the NE release and vasoconstriction responses at 1 min remained unaffected. The RNS-induced increases in ANG II-FR were completely abolished during infusion of enalaprilat. These results suggest that NE release on continuous RNS is enhanced by concomitantly formed ANG II, and this interaction depends on the time-related changes in intrarenal ANG II formation during RNS in the canine kidney. PMID- 10836715 TI - Evidence for vasoconstriction mediated by the endothelin-B receptor in domestic swine. AB - Endothelin-1 (ET-1), a potent vasoactive and mitogenic peptide, has been implicated in a number of cardiovascular diseases, including congestive heart failure, neointimal hyperplasia associated with restenosis, and hypertension. The vasoconstriction induced by ET-1 is thought to be mediated mainly by its action on ET(A) receptors on vascular smooth muscle cells. Recent studies have indicated that vasoconstriction also may be mediated by stimulation of an ET(B)-receptor subtype. Increased use of the pig as a cardiovascular model prompted us to examine the receptor profile in this species using ABT-627, a potent, nonpeptide antagonist of the ET(A) receptor. The precursor to ET-1, big ET-1 (0.02 nmol/kg/min), was infused intravenously in domestic swine, resulting in a sustained increase in mean blood pressure of 38 +/- 3 mm Hg. After stabilization of the pressor response, ABT-627 (0.1-10 microg/kg/min) or vehicle was infused for 30 min. Whereas vehicle infusion had no appreciable effect, a dose-related reversal of the pressor response to big ET-1 (11-100%) was observed by the end of the ABT-627 infusion. Blood samples were assayed for plasma concentrations of ABT 627; peak levels ranged from 9 +/- 2 to 937 +/- 168 ng/ml. In a separate group of pigs, the highest dose of ABT-627 produced only a modest reversal of the hypertensive response to an infusion of angiotensin II (300 ng/kg/min). Additional results indicate that the vasoconstrictor effects produced by sarafotoxin 6C (0.03 and 0.3 nmol/kg), an agonist of the ET(B) receptor, are not blocked by treatment with ABT-627 (10 microg/kg/min). However, complete blockade of the S6C response could be achieved using the ET(B) antagonist, A-192621 (0.33 mg/kg/min). Our results define the dose-response relation for the ET(A)-receptor antagonist ABT-627 in the vasculature of the domestic pig and suggest the presence of an ET(B)-receptor subtype that mediates vasoconstriction in this species. PMID- 10836716 TI - Nebivolol inhibits human aortic smooth muscle cell growth: effects on cell cycle regulatory proteins. AB - An enhanced vasoconstriction and vascular smooth muscle cell proliferation are involved in pathogenesis of hypertension. Beta3-blockers are effective for treatment of hypertensive patients. Recently the new beta1-receptor blocker nebivolol showed a different hemodynamic profile from those of other classic beta blockers. In this study we hypothesized that nebivolol may also have different effects on smooth muscle cell proliferation compared with other beta-blockers such as atenolol. Human aortic smooth muscle cells (SMCs) were cultured, and cell growth was determined by increase in cell number. Growth-signaling molecules such as mitogen-activated protein kinase (p42mapk) and S6-kinase (p70S6K) and cell cycle regulatory proteins (i.e., Cdk2, p27Kip1, and pRb) were analyzed by immunoblotting. In cultured human aortic SMCs, cell number was markedly increased in response to 5% fetal calf serum (FCS) over 6 days (87 +/- 11 x 10(3)/well), which was inhibited by nebivolol (10(-8)-10(-5) M; 25 +/- 2 x 10(3)/well; n = 6; p < 0.05), but not by atenolol. 5% FCS activated p42mapk, S6K, and Cdk2, but downregulated p27Kip1 and hyperphosphorylated pRb. Nebivolol prevented Cdk2 activation without influencing p42mapk, S6K, pRB, and p27Kip1. Thus, the new beta1-blocker nebivolol exhibits antiproliferative effect on human SMC through inactivation of Cdk2. This effect of nebivolol may have advantages over other beta-blockers in treatment of patients with cardiovascular disease. PMID- 10836717 TI - Serotonin reuptake inhibitor fluoxetine decreases arteriolar myogenic tone by reducing smooth muscle [Ca2+]i. AB - Previous studies showed that the serotonin reuptake inhibitor (SSRI) antidepressant fluoxetine (Prozac) dilates skeletal muscle and cerebral arterioles independent of the endothelium. We hypothesized that fluoxetine affects the contractile activity of arteriolar smooth muscle by interfering with Ca2+ signaling pathways. The effects of fluoxetine on pressure-induced tone of isolated rat skeletal muscle arterioles (approximately 110 microm) were investigated by videomicroscopy. Changes in smooth muscle [Ca2+]i were measured simultaneously by the fura-2 ratiometric method. Elevation of intraluminal pressure (from 20 to 120 mm Hg) increased (by approximately 20%) the smooth muscle calcium fluorescence ratio (R(Ca)) and resulted in a significant myogenic constriction (approximately 40%). Fluoxetine and nifedipine significantly decreased R(Ca) (by approximately 30%) and abolished pressure-induced arteriolar tone (EC50, 3.1 x 10(-6) and 6.0 x 10(-9) M, respectively). Constrictions to the L-type Ca2+ channel opener Bay K 8644 also were inhibited and abolished by increasing doses of fluoxetine (3 x 10(-6) and 10(-5) M, respectively). In the presence of 10(-5) M fluoxetine, a concentration that elicited submaximal (approximately 80%) dilation, elevation of extracellular Ca2+ concentration (from 2.5 to 15 mM) normalized R(Ca) and restored arteriolar myogenic tone. Thus, fluoxetine reduces [Ca2+]i and tone of arteriolar smooth muscle, likely by interfering with Ca2+ entry. We speculate that the "calcium antagonist" effect of fluoxetine may be an additional element in the therapeutic actions of this drug. PMID- 10836718 TI - SM-20550, a new Na+/H+ exchange inhibitor and its cardioprotective effect in ischemic/reperfused isolated rat hearts by preventing Ca2+-overload. AB - We investigated the effect of a newly synthesized compound, SM-20550 [N (aminoiminomethyl)-1,4-dimethyl-1H-indole-2-carboxamide methanesulfonic acid] on Na+/H+ or Na+/Ca2+ exchange activity in rat cardiomyocytes, and on radioligand binding with several channels or receptors in membrane preparations, and ischemia/reperfusion injury in isolated perfused rat hearts. In myocytes, SM 20550 concentration-dependently inhibited the recovery from acidosis induced by an NH4Cl prepulse in HCO3(-)-free solution. Its IC50 was 10(-8) M, which was 10 times lower than that of ethylisopropyl amiloride (EIPA). SM-20550 (10(-6) M) did not affect the Na+-dependent Ca2+ influx (Na+/Ca2+ exchange activity) in cardiomyocytes. In the radioligand binding assay, SM-20550 did not have affinity for K+ channel, beta-adrenoceptor, adenosine, angiotensin, or endothelin receptors, and had low affinity for Na+ and Ca2+ channels and alpha adrenoceptors, only at the concentrations of 10(-6)-10(-5) M. In perfused hearts exposed to 40 min of global ischemia and 20 min of reperfusion, SM-20550 (10(-8) 10(-7) M) significantly reduced the elevation of left ventricular end-diastolic pressure during reperfusion, improved the postischemic recovery of developed pressure, and prevented coronary perfusion pressure increase after reperfusion. Furthermore, SM-20550 reduced creatine phosphokinase release during reperfusion and prevented the abnormal gain of tissue Na+ and Ca2+ at the end of reperfusion. These results suggest that SM-20550 is a potent, highly specific Na+/H+ exchange inhibitor, which exerts a protective effect against myocardial ischemia/reperfusion injury. In addition, our data strongly support the hypothesis that Na+/H+ exchange plays an important role in the development of postischemic cardiac dysfunction, most likely by inducing Na+ and Ca2+ overload. PMID- 10836719 TI - Phenylephrine precontraction increases the sensitivity of rabbit femoral artery to serotonin by enabling 5-HT1-like receptors. AB - We used selective receptor antagonists to identify the receptors mediating the isometric contractile response to serotonin in control and phenylephrine (PHE) precontracted rabbit femoral artery rings. Serotonin, in the absence of PHE, elicited monophasic concentration-response curves (CRCs) early, but biphasic CRCs late in the course of the study. In the monophasic curves, the threshold and maximal concentrations were 10 and 1,000 microM, respectively. In biphasic CRCs, the threshold and maximal concentrations of the first phase were 0.03 and 3 microM, respectively. The respective values for the second phase were 10 and 1,000 microM. Prazosin, 0.1 microM, a selective alpha1-adrenoceptor antagonist, inhibited the monophasic curves, but only the second phase of the biphasic curves. Ritanserin, 0.01 microM, a selective 5-HT2A-receptor antagonist, shifted the first phase of the biphasic serotonin CRCs to the right but had little effect on the second phase. PHE increased the sensitivity of rabbit femoral artery response to serotonin. This amplified response to serotonin was antagonized by 0.01 microM GR 127935T, a selective 5-HT1B-receptor antagonist. The selective 5 HT1 agonist, sumatriptan, had no effect in control femoral arteries, but caused a concentration-dependent contraction after PHE precontraction. These results suggest that 5-HT1-like receptors are normally inactive or "silent" in the absence of PHE. However, in the presence of PHE, these receptors become enabled and mediate the amplified response to serotonin. The evidence also suggests that, in the absence of PHE, alpha1-adrenoceptors mediated the contractile response to serotonin in the monophasic CRCs. In the biphasic curves observed late in the study, the first phase was mediated by 5-HT2A receptors, and the second, by the alpha1-adrenergic receptors. PMID- 10836720 TI - Vascular and endothelial actions of inhibitors of substance P amidation. AB - Formation of mature active neuropeptides such as substance P (SP) from their glycine extended precursors entails alpha-amidation of peptide precursors by the sequential enzymatic action of peptidylglycine alpha-monooxygenase (PAM) and peptidylamidoglycolate lyase (PGL). We reported that these two enzymes that can produce mature active neuropeptides are present in cultured bovine aortic endothelial cells (BAECs). We hypothesize that alpha-amidation of peptides occurs in endothelial cells and that these peptides are critically involved in the overall regulation of cardiovascular function. In this study, this hypothesis was tested using specific amidation inhibitors to determine their effects on the actions of SP and its glycine-extended precursor (SP-Gly). We have found that SP and SP-Gly are equipotent in stimulating nitric oxide (NO) release by BAECs. At 10(-5) M, the specific inhibitors of PAM (4-phenyl-3-butenoic acid; PBA) and PGL (5-acetamido-2,4-diketo-6-phenyl-hexanoic acid and its methyl ester) reduced NO basal release by 40, 34, and 45%, respectively. They also reduced the production of NO induced by SP-Gly by 63, 68, and 69%, respectively, but had no effect on NO production in response to either SP or acetylcholine. SP and SP-Gly also were equipotent in relaxing rat aortic segments. The vasorelaxation to SP-Gly was endothelium dependent and inhibited by the NOS antagonist L-nitroarginine methyl ester (L-NAME), but it was not affected by inhibition of prostaglandin synthesis. Inhibitors of both PAM and PGL significantly reduced the vasorelaxing actions of SP-Gly, whereas responses to SP were not affected. A cumulative infusion of PBA into the femoral artery of rabbits, at final concentrations of 2.4, 24, and 240 microM for 20 min each, increased the vascular resistance (VR), indicating the tonic production of vasodilating amidated peptide(s). This effect was maximum at 60 min after infusion (20.5 +/- 4.7 vs. 8.2 +/- 0.7 mm Hg/ml/min; p < 0.05). These results suggest that endothelial cells can produce mature SP from its SP Gly precursor and that a product of peptide alpha-amidation tonically stimulates endothelial cell NO release to control vascular tone. PMID- 10836721 TI - Inotropic effects of diadenosine monophosphate (AP1A) in isolated human cardiac preparations. AB - Dependent on the number of phosphate residues, diadenosine polyphosphates (APnP) exert divergent inotropic effects in the human heart. We studied the inotropic effects of the smallest member of this family, diadenosine monophosphate (AP1A). Force of contraction was measured in an isometric setup in isolated electrically driven (0.5 Hz) preparations from human atria. AP1A exerted a concentration dependent negative inotropic effect. The IC50 value was 20.2 microM and the IC50 value was 3.1 microM (n = 5-8). At 100 microM AP1A, force of contraction declined to 50% of the predrug value after 2.5 +/- 0.5 min of incubation (n = 8). AP1A antagonized the positive inotropic effect of the beta-adrenoceptor agonist isoprenaline (10 nM). For 100 microM AP1A, the time to 50% of the predrug force in the presence of isoprenaline amounted to 2.3 +/- 0.2 min (n = 5). The positive inotropic and lusitropic effects of isoprenaline were antagonized by AP1A. The direct (AP1A alone) and indirect (AP1A in the presence of isoprenaline) negative inotropic effects of AP1A were blocked by the A1-adenosine receptor antagonist 1,3-dipropyl-cyclopentyl-xanthine (DPCPX, 0.3 microM). The inotropic effect of AP1A was not blocked by adenosine deaminase. In conclusion, AP1A exerts indirect and direct negative inotropic effects in the human heart through A1-adenosine receptors. These effects might protect the heart against excessive beta adrenergic stimulation. PMID- 10836722 TI - Amlodipine inhibits expression of matrix metalloproteinase-1 and its inhibitor in human vascular endothelial cells. AB - Matrix metalloproteinase-1 (MMP-1) may play an important role in the pathogenesis of atherosclerosis and atherosclerotic plaque rupture. We investigated the effect of the calcium channel blockers amlodipine and nifedipine on the expression of MMP-1 and tissue inhibitor of metalloproteinase-1 (TIMP-1) in endothelial cells (ECs). MMP-1 and TIMP-1 levels in conditioned media of human vascular ECs were measured by enzyme-linked immunosorbent assay. Collagenolytic activity was determined by fluorescence-labeled collagen digestion. The addition of interleukin-1beta (IL-1beta) increased MMP-1 levels in the culture media of ECs. Amlodipine, but not nifedipine, significantly decreased MMP-1 levels in IL-1beta stimulated ECs. TIMP-1 levels also were significantly increased by IL-1beta, and its expression was slightly decreased by amlodipine, not by nifedipine. Amlodipine significantly inhibited collagenolytic activity in the culture media of IL-1beta-stimulated ECs, whereas nifedipine showed no significant effect on the activity. Our findings revealed that amlodipine, but not nifedipine, inhibits IL-1beta-induced MMP-1 expression in human ECs. PMID- 10836723 TI - Effects of short- and long-term exercise on urinary cGMP excretion in healthy subjects and in patients with coronary artery disease. AB - The possibility that systemic formation of cyclic guanosine monophosphate (cGMP) could reflect the level of cardiovascular fitness was investigated. The relations between physical activity and systemic formation of cGMP were evaluated in healthy volunteers and in patients with coronary artery disease (CAD). No significant differences were observed in the basal urinary excretion of cGMP in highly trained runners, sedentary subjects, and in patients with CAD, despite the large differences in aerobic exercise training between groups. In addition, the basal levels of cGMP in CAD patients failed to increase after a 12-week cardiac rehabilitation program. Short-term exercise, on the other hand, was associated with significant increases in urinary cGMP excretion. A 42-km marathon increased urinary cGMP excretion by 272%. The 15-km race increased urinary cGMP excretion by 330%. In CAD patients, 30 min of supervised exercise on a treadmill, at 80% of patient's maximal heart rate, induced a 60% increase in urinary cGMP, which returned to preexercise levels 90 min after termination of the exercise. Completion of the 12-week cardiac rehabilitation program improved exercise capacity and the magnitude of increase in cGMP levels induced by short-term treadmill exercise. Our findings suggest that cGMP increases during and shortly after short-term exercise and that the magnitude of the increase seems dependent on the intensity of the exercise and on physical fitness. Exercise training in healthy subjects and in CAD patients enhanced the amount of cGMP produced during short-term exercise, which might be responsible for some of the protective cardiovascular actions of exercise. The short half-life of cGMP may explain why the basal resting levels of cGMP are not appropriate predictors of a subject's physical fitness. PMID- 10836724 TI - Acute effects of a single low oral dose of pimobendan on left ventricular systolic and diastolic function in patients with congestive heart failure. AB - A recent long-term multicenter trial has shown that pimobendan is more effective when administered in low doses. However, no data are available concerning the effect of a low dose of pimobendan on the systolic and diastolic pressure-volume relations in patients with heart failure. Therefore we examined the effects of a single low dose of oral pimobendan, a calcium sensitizer, on systolic and diastolic hemodynamics in patients with cardiomyopathy and congestive heart failure. We measured the left ventricular (LV) pressure-volume relations using a conductance catheter with a micromanometer tip in 10 patients with chronic congestive heart failure resulting from idiopathic cardiomyopathy before and 45 and 90 min after administration of a single oral dose of 2.5 mg of pimobendan. End-systolic elastance was used as an index of LV contractility and was measured during transient occlusion of the inferior vena cava. End-systolic elastance increased significantly by 25% at 45 min (p < 0.05) and by 55% at 90 min (p < 0.01) without an increase in myocardial oxygen consumption. The inotropic effect was accompanied by improved ventriculoarterial coupling. This effect was attenuated in patients with severely impaired myocardial contractility. LV relaxation, assessed by the time constant of isovolumic pressure decay (T(1/2)), was significantly shortened at 90 min (from 47.7 +/- 1.9 to 41.2 +/- 1.7 ms; p < 0.01), although it remained unchanged at 45 min. The diastolic pressure-volume relation showed a leftward and downward shift in all patients. These results indicate that low-dose oral pimobendan had favorable short-term inotropic and lusitropic effects in patients with congestive heart failure caused by idiopathic dilated cardiomyopathy, and may thus be a useful alternative to traditional agents. Further study in a large-scale trial is merited. PMID- 10836725 TI - Comparative effects of angiotensin II AT-1-type receptor antagonists in vitro on human platelet activation. AB - A recent study has shown that losartan, an AT-1-receptor antagonist, interacts with thromboxane A2 (TxA2)/prostaglandin H2 (PGH2) receptors in human platelets. The aim of this study was to analyze the ability of different angiotensin II (Ang II) AT-1-receptor antagonists to inhibit TxA2-dependent human platelet activation. Platelets were obtained from healthy volunteers. Platelets were stimulated with the TxA2 analogue, U46619 (10(-6) M). U46619-stimulated platelet activation was significantly reduced by both losartan and irbesartan in a dose dependent manner. Only maximal doses of valsartan (5 x 10(-6) M) and the main metabolite of losartan, EXP3174 (5 x 10(-6) M), reduced U46619-induced platelet activation. Whereas the active form of candesartan cilexetil (candesartan, CV 11974) failed to modify platelet activation involved by TxA2, telmisartan showed a higher effect than valsartan and EXP3174 but lower than either losartan and irbesartan. Losartan or irbesartan reduced the binding of [3H]-U46619 to platelets, an effect that was observed with lower ability with the other AT-1 antagonists. Although platelets expressed AT-1-type receptors, exogenous Ang II did not modify platelet activation. This effect was not modified by blocking the AT-2 receptor with PD123319. These results suggest that some AT-1-receptor antagonists reduce TxA2-dependent activation independent of Ang II involvement. PMID- 10836726 TI - Salutary antiarrhythmic effect of combining a K channel blocker and a beta blocker in a canine model of 7-day-old myocardial infarction. AB - We sought to examine whether the antiarrhythmic effect of E4031 (E), or I(Kr) channel blocker, is affected by beta-adrenergic stimulation using isoproterenol (Iso) or by beta-adrenergic blockade (betaB) using, ONO1101, in a canine myocardial infarction model. Electrophysiologic studies were performed in 10 dogs with 7-day-old myocardial infarctions. Local QT intervals were measured at 47 sites on the infarcted myocardium using a mapping electrode. QT dispersion (QTd), as defined by the coefficient of variation of QT intervals, was obtained. Inducibility of ventricular arrhythmias was examined by programmed stimulation. These procedures were repeated during administration of E, E + Iso, and E + betaB. The effect of prolonging local QT intervals by E was counteracted by Iso, and was accentuated by betaB. The amount of prolongation was dependent on the baseline QT intervals, and QTd showed a tendency to decrease with E, to increase with E + Iso, and significantly decreased with E + betaB. Ventricular tachyarrhythmias were induced in a half of dogs with E + Iso, but were not induced with E + betaB. In the presence of adrenergic activation, I(Kr) blockers may exhibit a decreased antiarrhythmic effect. Beneficial synergism can be expected when an I(Kr) blocker is combined with a beta-blocker in the subacute phase of myocardial infarction. PMID- 10836727 TI - Effects of intravenous nicorandil on coronary circulation in humans: plasma concentration and action mechanism. AB - We investigated the cardiovascular profile of nicorandil, an antianginal agent, in humans. Pharmacologically, nicorandil acts as both an adenosine triphosphate (ATP)-sensitive K+ (K(ATP)) channel opener and a nitrate. We examined which of these mechanistic components has a predominant vasodilatory effect at clinical doses. Fourteen patients underwent cardiac catheterization. The effects of the continuous intravenous infusion of nicorandil (12 mg/45 min) were examined in angiographically normal coronary arteries. Coronary vascular resistance was calculated from coronary artery diameter and coronary blood flow velocity measured using an intravascular Doppler catheter. We compared the hemodynamic responses to nicorandil with those to the intracoronary injection of nitroglycerin (250 microg) and papaverine (12 mg). The epicardial coronary arteries responded to nicorandil at the lowest plasma concentration examined (dilation of +14.0 +/- 3.3% at approximately 170 ng/ml), whereas dilation of the coronary resistance arteries (i.e., a decrease in coronary vascular resistance) took place only at higher concentrations (>200 ng/ml). Nitroglycerin caused no further changes in coronary artery diameter or coronary vascular resistance. Papaverine caused no further increase in coronary artery diameter, but markedly decreased coronary vascular resistance (1.6 +/- 0.3 to 0.4 +/- 0.1 mm Hg/ml/min; p < 0.05). Nicorandil significantly decreased pulmonary capillary wedge pressure (i.e., reduced cardiac preload) at a plasma level of >200 ng/ml, but did not change either systemic or pulmonary vascular resistance. Thus nicorandil preferentially dilated epicardial coronary arteries rather than coronary resistance arteries, and had a stronger effect on preload than on afterload. These changes in human coronary hemodynamics suggest that the nitrate actions of nicorandil as a coronary vasodilator predominate over those as a K(ATP) opener. PMID- 10836728 TI - Antihypertensive efficacy of manidipine and enalapril in hypertensive diabetic patients. AB - Recent studies showed that in diabetic hypertensive patients, administration of angiotensin-converting enzyme (ACE)-inhibitors or calcium antagonists can effectively lower blood pressure (BP) and prevent diabetes-related cardiovascular complications with no adverse metabolic effects. We sought to assess the antihypertensive and metabolic effects of the new dihydropyridine calcium antagonist manidipine (M) in patients with diabetes mellitus and essential hypertension as compared with the ACE inhibitor enalapril (E). After 3 weeks of placebo, 101 (62 men; age range, 34-72 years) hypertensives with type II diabetes mellitus were randomized to M 10-20 mg or E 10-20 mg, od, for 24 weeks. At the end of the placebo period and the active-treatment phase, BP was measured with a mercury sphygmomanometer (office, O) and over the 24 h by ambulatory (A) monitoring. ABP recordings were analyzed to obtain 24-h, day (6 a.m. to midnight), and night (midnight to 6 a.m.) average systolic (S) and diastolic (D) BP and heart rate (HR) values. Homogeneity of the antihypertensive effect over the 24 h was assessed by the smoothness index [SI: i.e., the ratio between the average of the 24 hourly BP changes after treatment and the corresponding standard deviation (the higher the SI, the more uniform is the BP control by treatment over the 24 h]. The O SBP and DBP were significantly (p < 0.01) and similarly reduced by M (16 +/- 10 and 13 +/- 6 mm Hg, n = 49) and E (15 +/- 10 and 13 +/- 6 mm Hg, n = 45). The percentage of patients whose O DBP was reduced < or = 85 mm Hg (i.e., the value indicated to be the optimal DBP goal in diabetic hypertensives) was similar for M (37%) and E (40%). The reduction of 24-h BP also was similar between M (n = 38) and E (n = 38) for both drugs (systolic, 6 +/- 11 and 8 +/- 10 mm Hg; diastolic, 5 +/- 8 and 5 +/- 7; NS, M vs. E). The antihypertensive effect was distributed in a similar homogeneous fashion throughout the dosing interval, as shown by the similar SI values (M, 0.6 +/- 1.2 for SBP and 0.6 +/- 0.9 for DBP; E, 0.6 +/- 0.8 for SBP and 0.5 +/- 0.7 for DBP; NS, M vs. E). O and A HR were unchanged by either treatment. Markers of glucose and lipid metabolism and renal function were not significantly modified by treatment both with M and with E. In the diabetic hypertensives, M was as effective and metabolically neutral as the ACE-inhibitor E. PMID- 10836729 TI - Differential effects of imidapril and candesartan cilexetil on plasminogen activator inhibitor-1 expression induced by prolonged inhibition of nitric oxide synthesis in rat hearts. AB - We investigated effects of the angiotensin-converting enzyme (ACE) inhibitor imidapril and the angiotensin II type 1 (AT1) antagonist candesartan cilexetil on cardiac plasminogen activator inhibitor-1 (PAI-1) expression in rats. Cardiac PAI 1 mRNA levels were increased after a 7-day treatment with the nitric oxide (NO) synthesis inhibitor N(omega)-nitro-L-arginine methyl ester (L-NAME). PAI-1 immunoreactivity was increased in the coronary arteries. Treatment with imidapril significantly prevented the L-NAME-induced increase in the gene expression and immunoreactivity of PAI-1, but candesartan cilexetil showed no such effect. This study provides the first evidence of differential effects of ACE inhibition and AT1 antagonism on cardiac PAI-1 expression in vivo. PMID- 10836730 TI - Additive hypotensive effect of angiotensin-converting enzyme inhibition and angiotensin-receptor antagonism in essential hypertension. AB - The study was designed to assess the antihypertensive effect of combined angiotensin-converting enzyme (ACE) inhibition and angiotensin II type 1 receptor (AT1) antagonism in patients with essential hypertension. Twenty patients with uncontrolled ambulatory diastolic blood pressure (BP) after 6 weeks of ACE inhibitor monotherapy (benazepril, 20 mg, o.d.) were randomized to receive double blind valsartan, 80 mg, o.d. (AT1 antagonist) or matching placebo for 5 weeks while continuing to receive background benazepril. Then patients crossed over to the alternative regimen for a second 5-week period. The 24-h ambulatory BP was monitored on the final day of the benazepril monotherapy period and on the final day of each double-blind treatment period. Valsartan added to benazepril produced a significant antihypertensive effect with a benefit over placebo of 6.5 +/- 12.6/4.5 +/- 8.0 mm Hg (systolic/diastolic) for average awake ambulatory BP (p < 0.05), 7.1 +/- 9.4/5.6 +/- 6.5 mm Hg for asleep BP (p < 0.01), and 6.8 +/- 9.7/4.9 +/- 6.8 mm Hg for average 24-h ambulatory BP (p < 0.01). Pulse rate was unaffected. Plasma active renin was higher on the benazepril-valsartan combination compared with benazepril-placebo (p < 0.05). There was no change in routine biochemical variables when valsartan was added to benazepril. Six patients reported mild dizziness or fatigue (three also with placebo). These data suggest that in hypertensive patients uncontrolled with an ACE inhibitor, the addition of an AT1 antagonist provides a powerful and safe antihypertensive drug combination. PMID- 10836731 TI - Distinct action of aranidipine and its active metabolite on renal arterioles, with special reference to renal protection. AB - Aranidipine, a newly developed calcium antagonist, possesses unique pharmacologic characteristics in that its metabolite (M-1) still has antihypertensive action. We examined the effects of both agents on renal microcirculation using the isolated perfused hydronephrotic rat kidney. During norepinephrine-induced constriction, the addition of aranidipine dilated both afferent and efferent arterioles in a dose-dependent manner; at 10(-6) M, 83 +/- 6% and 90 +/- 6% reversal, respectively. In contrast, its active metabolite exerted dilator action predominantly on the afferent arteriole (79 +/- 4% vs. 44 +/- 17% at 10(-6) M for afferent and efferent arterioles, respectively). We further examined the long term (8 weeks) effect of these agents on the development of renal injury in salt loaded subtotally nephrectomized spontaneously hypertensive rats. Both aranidipine and M-1 reduced blood pressure by a similar magnitude. The decreases in proteinuria were observed in the aranidipine-treated group at weeks 6, 8, and 10, whereas in the M-1 group, significant reduction was attained only at week 6. Histopathologic examination revealed that both treatments improved glomerular and arteriolar sclerosis. Glomerular sclerosis, however, was less pronounced in the aranidipine-treated group than in the M-1 group. In conclusion, aranidipine has dilator action on both arterioles, whereas M-1 caused predominant dilation of afferent arterioles. Such metabolic changes may constitute a determinant of efferent arteriolar action of the calcium antagonist. PMID- 10836732 TI - Inhibitors of ischemic preconditioning do not attenuate Na+/H+ exchange inhibitor mediated cardioprotection. AB - Pharmacologic inhibition of the K(ATP) channel with sulfonylureas or the adenosine receptor with methylxanthines has been shown to attenuate ischemic preconditioning (IPC). Both classes of compounds are widely used clinically, and several reports have demonstrated adverse outcomes in patients taking sulfonylureas. Recently inhibition of the sodium/hydrogen exchanger isozyme-1 (NHE-1) has been shown to be equal to IPC at providing myocardial protection in dogs and may be an alternative to IPC in patients taking sulfonylureas or methylxanthines. However, no experiments have examined the pharmacologic overlap between IPC and NHE-1 inhibitor-mediated cardioprotection in dogs. With an in vivo canine infarct model in which the left anterior descending coronary artery was occluded for 60 min and reperfused for 3 h, neither the K(ATP) channel antagonist glibenclamide nor the adenosine-receptor antagonist PD 115199 attenuated NHE-1 inhibitor-mediated reduction in infarct size expressed as a percentage of the area at risk produced by EMD 85131 (Control, 24.2 +/- 3.6%; EMD 85131, 6.4 +/- 2.3%; PD 115199 + EMD 85131, 6.6 +/- 2.4%; glibenclamide + EMD 85131, 3.5 +/- 1.2%). NHE-1 inhibition and IPC do not overlap pharmacologically, and NHE-1 inhibition may be an alternative for cardioprotection in patients taking sulfonylureas or methylxanthines. PMID- 10836733 TI - Acid and base catalyzed intramolecular cyclizations of N-benzoylthiocarbamoyl acetals. AB - Acid and base catalyzed intramolecular cyclizations of N-benzoylthioureidoacetal, containing four functional groups adjacent to thiourea such as benzocarbamoyl, acetal, thioure and amide, were investigated. The condensation reaction of N benzoyl thiocarbamoylglycine amide in the presence of 10% aqueous NaOH provided 1 (2,2-dimethoxy)ethyl-imidazolidine-2-thione exclusively. In the presence of pyridine, it was transformed to 2-thiohydantoin. N-Benzoyl thiocarbamoyl glycine amide was completely transformed to an iminothiazolidine exclusively in the presence of Lewis acid such as borontrifluoride etherate or trimethylsilyl iodide. 1-(2,2-Dimethoxy)ethyl-imidazolidine-2-thione was transformed to imidazole[2,1-b]thiazole and pyrazino[5,1-a]imidazole in the presence of BF3.Et2O and formic acid, respectively. PMID- 10836734 TI - Synthesis and characterization of stereospecific 1-propargyl-2-(dimethoxymethyl) 1-cyclohexanols. AB - Stereochemical isomers with hydroxy groups were synthesized by reacting 2 (dimethoxymethyl)cyclohexanone with propargylmagnesium bromide. The stereo chemical structures were identified by NMR spectral interpretation and the geometry optimization. To assist the NMR interpretation, geometry optimization based on semi-empirical AM1 and PM3 methods was applied. Throughout this study, the structures of the two isomers were all determined and 1H and 13C NMR spectra were fully assigned. It was proven that the less polar isomer is an axial alcohol and the more polar one is an equatorial alcohol. PMID- 10836735 TI - Synthesis of 4,6-dichloro-3-[(1-N-arylaminocarbonyl)-hydrazono]-1,3-dihydro indole-2 -one as a potential NMDA receptor glycine site antagonist. AB - A synthetic procedure for the preparation of indole-2,3-dione derivatives 6 as a potential NMDA receptor glycine site antagonist with improved pharmacological profile compared with 2-carboxyindole derivative 5, starting from readily available 3,5-dichloroaniline (7), is described. PMID- 10836736 TI - Synthesis and biological activity of aspirin derivatives. AB - Aspirin has been widely used as analgesic and anti-inflammatory drug. Recently, it was elucidated that aspirin have anti-coaggregatory effect in low dose. This study was carried out to investigate the synthesis of aspirin derivatives from aspirin and aromatic compound of antioxidant and its biological activities. Synthesis of aspirin derivatives was prepared by esterification in the presence of 1,1-carbonyldiimidazole. Biological activities was examined using effect of anti-coagulant on bleeding time, effect of antioxidant and effect of anti platelet aggregation. As a result, SJ-101 showed strong antioxidative activity and anti-coagulant activity among four compounds. Anti-platelet aggregation of SJ 101 was examined by collagen, ADP, PAF method. SJ-101 exhibited more stronger activity to aspirin at collagen aggregation reaction. These finding demonstrates that SJ-101 is useful as care drug of aging and old-disease because of its has antioxidant activity, anti-coagulant activity and anti-platelet activity. PMID- 10836737 TI - Synthesis and cytotoxic effects of deoxy-tomentellin. AB - Cannabigerol (1, CBG), methyl 4-[(2E)-3,7-dimethyl-2,6-octadienyl)oxy]-3 methoxybenzoate (2, DTM), 5-fluorouracil (3, FU) as a reference, and cannabidiol (4, CBD) were tested for their growth inhibitory effects against KB(ATCC NO, OCL 17) cell lines using two different assays, the 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl-2H-tetrazoliumbromide (MTT) assay and the sulforhod-amine B protein (SRB) assay. These compounds showed inhibitory activity in vitro in the micromolar range against KB cell lines. In general, the antitumor activities of these compounds (1, 2, 3 and 4) were dose-dependent over the micromolar concentration range of 1 to 100 M. The comparison of IC50 values of these compounds in tumor cell lines showed that their susceptibility to these compounds decreases in the following order: DTM > CBD > 5-FU > CBG by MTT assay and DTM = CBD > 5-FU > CBG by SRB assay. CBG 1, DTM 2, 5-FU 3, and CBD 4 were tested for their cytotoxic effects on NIH 3T3 fibroblasts using two different assays, the MTT assay and SRB assay. These compounds exhibited potent cytotoxic activities in vitro in the micromolar range against NIH 3T3 fibroblasts. In general, the cytotoxic activities of these compounds (1, 2, 3 and 4) were dose-dependent over the micromolar concentration range of 1 to 100 M. The comparison of CD50 values of these compounds in NIH 3T3 fibroblasts shows that their susceptibility to these compounds in decreases the following order(:) CBD > 5-FU > DTM > CBG by MTT assay, CBD > 5-FU > CBG > DTM by SRB assay. These results suggest that DTM 2 has the most growth-inhibitory activity against KB cell lines. PMID- 10836738 TI - Synthesis of new uracil-5-sulphonamide-p-phenyl derivatives and their effect on Biomphalaria alexandrina snail's nucleoproteins. AB - In continuation of the previous work (Fathalla, 1992) on the synthesis of some heterocycles containing uracil moiety, we report herein the incorporation of uracil moiety into cyanopyridine thione, thiosemecarbazone, semicarbazone, cyanopyridine, aminocyano pyridine, isoxazoline, pyrazoline, pyrimidine, triazolo pyrimidine, pyran, selena and thiazole derivatives which might modify their biological activities. The biological studies revealed that the chemical compound III f showed high molluscicdal activity than other compounds. The profile of the nucleoprotein extracted from chemically (compound IIIc, e, f and g) treated or UV irradiated B. alexandrina snails did not show appreciable differences when compared to non-treated (native) snails by using SDS-PAGE, where no obvious qualitative or quantitative differences were observed. Immunization of experimental animals with the nucleoprotein extracted from native, chemically (compound III f & g) treated or physically treated B. alexandrina snails induced significant protection against challenge with normal S. mansoni cercariae, as compared to the non-immunized challenged control. As well as, a decrease in the number of granuloma formation and the size range of granuloma was also observed in immunized animals. It is concluded that, compounds III f and g have a potent molluscicidal activity. They also induced chemical modification comparable to that induced by physical treatment in the snail's nucleoprotein, which could possibly be used in immunization against S. mansoni infection. PMID- 10836740 TI - Lipid peroxidation inhibitory activity of some constituents isolated from the stem bark of Eucalyptus globulus. AB - Twelve compounds with lipid peroxidation inhibitory activity were isolated from the stem bark of E. globulus. Their structures were assigned as a new aromatic monoterpene (1) and eleven known compounds, pinoresinol (2), vomifoliol (3), 3,4,5-trimethoxyphenol 1-O-beta-D-(6'-O-galloyl)glucopyranoside (4), methyl gallate (5), rhamnazin (6), rhamnetin (7), eriodictyol (8), quercetin (9), taxifolin (10), engelitin (11), and catechin (12) on the basis of UV, mass, and NMR spectroscopic analyses. These compounds except vomifoliol significantly inhibited lipid peroxidation in rat liver microsome. PMID- 10836739 TI - Synthesis and characterization of oligonucleotides containing site-specific bulky N2-aralkylated guanines and N6-aralkylated adenines. AB - 7-Bromomethylbenz[a]anthracene is a known mutagen and carcinogen. The two major DNA adducts produced by this carcinogen, i.e., N2-(benz[a]anthracen-7-ylmethyl) 2'-deoxyguanosine (2, b[a]a2G) and N6-(benz[a]anthracen-7-ylmethyl)-2' deoxyadenosine (4, b[a]a6A), as well as the simpler benzylated analogs, N2-benzyl 2'-deoxyguanosine (1, bn2G) and N6-benzyl-2'-deoxyadenosine (3, bn6A), were prepared by direct aralkylation of 2'-deoxyguanosine and 2'-deoxyadenosine. To determine the site-specific mutagenicity of these bulky exocyclic amino substituted adducts, the suitably protected nucleosides were incorporated into 16 base oligodeoxyribonucleotides in place of a normal guanine or adenine residues which respectively are part of the ATG initiation codon for the lac Z' alpha complementation gene by using an in situ activation approach and automated phosphite triester synthetic methods. The base composition and the incorporation of the bulky adducts into synthetic oligonucleotides were characterized after purification of the modified oligonucleotides by enzymatic digestion and HPLC analysis. PMID- 10836741 TI - Cytotoxic peroxides from Artemisia stolonifera. AB - Two sesquiterpene endoperoxides, 1S, 4R, 6R-1, 4-endoperoxy-bisabola-2, 10-diene (I), 1R, 4S, 6R-1, 4-endoperoxy-bisabola-2, 10-diene (II), and a sesquiterpene hydroperoxide, 1beta-hydroperoxygermacra-4 (15), 5, 10 (14)-triene (III) were isolated from the aerial parts of Artemisia stolonifera (Compositae). Their chemical structures were assigned by spectral evidences. Compounds I and II exhibited cytotoxicity against five human tumor cell lines with their ED50 values ranging from 0.20 to 5.43 microg/ml and from <0.1 to 0.87 microg/ml, respectively. PMID- 10836742 TI - Cytotoxic triterpenes from Crataegus pinnatifida. AB - Bioassay-guided fractionation of Crataegus pinnatifida (Rosaceae) gave two cytotoxic ursane-type triterpenes which were identified as uvaol (1) and ursolic acid (2) by physicochemical and spectroscopic methods. 3-Oxo-ursolic acid (3) was synthesized from ursolic acid (2) by Jones method. The cytotoxic activities of these compounds were tested against murine L1210 and human cancer cell lines (A549, SK-OV-3, SK-MEL-2, XF498, and HCT15) in vitro. Compounds 1 and 2 showed moderate cytotoxicities against L1210, whereas they showed weak activities against human cancer cell lines. However, compound 3 exhibited potent cytotoxic activities both in murine and in human cancer cell lines. PMID- 10836743 TI - A new stilbene diglycoside from Rheum undulatum. AB - A new stilbene diglycoside, piceatannol-3, 4'-O-beta-D-diglucopyranoside (I), together with desoxyrhaponticin (II), emodin-1-O-beta-D-glucopyranoside (III), and physcion-8-O-b-D-glucopyranoside (IV), were isolated from the rhizomes of cultivated Korean rhubarb rhizomes (Rheum undulatum), Jong DaeWhang, and the structures of I-IV were identified on the basis of chemical and spectral evidences. PMID- 10836744 TI - Modifier effects on supercritical CO2 extraction efficiency of cephalotaxine from Cephalotaxus wilsoniana leaves. AB - The effects of modifiers such as methanol, water, diethylamine in methanol (10 v/v %), and diethylamine in water (10 v/v %) were investigated at three different concentrations (1, 5, and 10 v/v %) of the modifiers in supercritical CO2 (SC CO2) in order to enhance the supercritical fluid extraction (SFE) efficiency of cephalotaxine from Cephalotaxus wilsoniana leaves. Among the modifiers employed, methanol basified with diethylamine was found to greatly enhance the extraction efficiency relative to any other modifiers employed. The results suggest that cephalotaxine in plant matrices may be readily changed from SC-CO2-insoluble salt to SC-CO2-soluble free base by basified modifiers. In addition, SC-CO2 modified with basified methanol could enhance the extraction efficiency of cephalotaxine more than 30% when compared to the conventional organic solvent extraction. PMID- 10836745 TI - In vitro cytotoxic effect of N-(phosphonacetyl)-L-aspartic acid in liposome against C-26 murine colon carcinoma. AB - We have investigated the in vitro cytotoxic effect of liposome-encapsulated N (phosphonacetyl)-L-aspartic acid (PALA) against C-26 murine colon cancer cells, and have compared it in this regard to free PALA. Three different PALA-containing liposomal formulations using distearoylphosphatidylcholine (DSPC), distearoylphosphatidylglycerol (DSPG), and polyethyleneglycol-derivatized distearoylphosphatidylethanolamine (PEG-DSPE) were made and their cytotoxicity was measured. In 72 hr continuous exposure experiment with C-26 cells, the 50% growth inhibitory concentration (IC50) of DSPG-PALA liposome formulation was 0.09 microM, which showed about 65-fold more potent than unencapsulated free PALA (5.1 microM). Similar degree of increase in cytotoxicity was also observed in 1 hr exposure experiment. However, the IC50 of PEG-DSPE-PALA liposome and DSPC-PALA liposome were 10.7 microM and 11.8 microM, respectively, which showed slightly less potent than unencapsulated free PALA. Physical characteristics of PALA liposomes, such as the size and drug:lipid ratio were also determined. In conclusion, negatively-charged DSPG-PALA liposome showed the highest cytotoxic effect among tested on the C-26 cells in vitro. PMID- 10836746 TI - Biotransformation of glycyrrhizin by human intestinal bacteria and its relation to biological activities. AB - The relationship between the metabolites of glycyrrhizin (18beta-glycyrrhetinic acid-3-O-beta-D-glucuronopyranosyl-(1-->2)-beta-D-glucuronide, GL) and their biological activities was investigated. By human intestinal microflora, GL was metabolized to 18beta-glycyrrhetinic acid (GA) as a main product and to 18beta glycyrrhetinic acid-3-O-beta-D-glucuronide (GAMG) as a minor product. The former reaction was catalyzed by Eubacterium L-8 and the latter was by Streptococcus LJ 22. Among GL and its metabolites, GA and GAMG had more potent in vitro anti platelet aggregation activity than GL. GA also showed the most potent cytotoxicity against tumor cell lines and the potent inhibitory activity on rotavirus infection as well as growth of Helicobacter pylori. GAMG, the minor metabolite of GL, was the sweetest. PMID- 10836747 TI - Enantioselective determination of cetirizine in human urine by HPLC. AB - In order to study the simultaneous determination of (+)- and (-)-cetirizine in human urine we have developed a chiral separation method by HPLC. A chiral stationary phase of alpha1-acidglycoprotein, the AGP-CSP, was used to separate the enantiomers. The pH of the phosphate buffer, as well as the content of the organic modifier in the mobile phase, markedly affected the chromatographic separation of (+)- and (-)-cetirizine. A mobile phase of 10 mmol/l phosphate buffer (pH 7.0)-acetonitrile (95: 5, v/v) was used for the urine assays. Ultraviolet absorption was monitored at 230 nm and roxatidine was employed as the internal standard for quantification. (+)-Cetirizine, (-)-cetirizine and the internal standard were eluted at retention times of 12, 16, and 32 mins, respectively. The detection limit for cetirizine enantiomers was 400 ng/ml of urine. A pharmacokinetic study was conducted with the help of 5 healthy female volunteers who were administered with a single oral dose of racemic cetirizine (20 mg). The peak area ratios provided by the cetirizine enantiomers were linear (r>0.997) over a concentration range of 2.5-200 microg/ml. The peak of the excreted cetirizine enantiomers appeared in the urine sample during the period of 1-2 hrs following the administration of the oral dose. The excreted level of (+) cetirizine was slightly higher than (-)-cetirizine but the difference was not statistically significant. However, this method appears to have applications for enantioselective pharmacokinetic studies of racemic drugs. PMID- 10836748 TI - Isolation and characterization of proteoglycan derived from human placenta and its biological activities. AB - Chondroitin sulfates proteoglycans were isolated from human placenta. For the identification of enzymatic digestion products of isolated proteoglycan, strong anion, exchange-high performance liquid chromatography (SAX-HPLC) was performed. By the action of chondroitin ABC and chondroitin B lyase, three unsaturated disaccharides 2-acetamide-2-deoxy-3-O-(beta-D-gluco-4-enepyranosyluronic acid)-D galactose (deltaDi-OS), 2-acetamide-2-deoxy-3-O-(beta-D-gluco-4 enepyranosyluronic acid)-6-O-sulfo-D-galactose (deltaDi-6S) and 2-acetamide-2 deoxy-3-O-(beta-D-gluco-4-enepyranosyluronic acid)-4-O-sulfo-D-galactose (deltaDi 4S) were produced from the human placenta proteoglycan. The anticoagulant activity of chondroitin sulfate proteoglycan was evaluated by activated partial thromboplastin time (aPTT) assay and thrombin time (TT) assay. The clotting times of aPTT and TT were increased from 72 to 144 sec and 19 to 27 sec, respectively. The immuno-modulating activity of chondroitin sulfate proteoglycan was examined by cell proliferation assay and these results suggest that it may play a role in suppression of the function of immune-related cells. PMID- 10836749 TI - Characterization of the pcbE gene encoding 2-hydroxypenta-2,4-dienoate hydratase in Pseudomonas sp. DJ-12. AB - Nucleotide sequence extending 2,3-dihydroxybiphenyl 1,2-dioxygenase gene (pcbC) and 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoate hydrolase gene (pcbD) of Pseudomonas sp. DJ-12 was previously analyzed and the two genes were present in the order of pcbD-pcbC preceded by a promoter from Pseudomonas sp. DJ-12. In this study, a 3.8-kb nucleotide sequence located downstream of the pcbC gene was analyzed to have three open reading frames (ORFs) that are designated as orf1, pcbE and orf2 genes. All of the ORFs were preceded by each ribosome-binding sequence of 5-GGAXA-3 (X=G or A). However, no promoter-like sequence and transcription terminator sequence were found in the analyzed region, downstream of pcbC gene. Therefore, the gene cluster appeared to be present in the order of pcbD-pcbC-orf1-pcbE-orf2 as an operon, which is unique organization characterized so far in biphenyl- and PCB-degrading bacteria. The orf1 gene was composed of 1,224 base pairs which can encode a polypeptide of molecular weight 44,950 containing 405 amino acid residues. A deduced amino acid sequence of the orf1 gene product exhibited 21-33% identity with those of indole dioxygenase and phenol hydroxylase components. The pcbE gene was composed of 783 base pairs encoding 2-hydroxypenta-2,4-dienoate hydratase involved in the 4-chlorobiphenyl catabolism. The orf2 gene was composed of 1,017 base pairs encoding a polypeptide of molecular weight 37,378 containing 338 amino acid residues. A deduced amino acid sequence of the orf2 gene product exhibited 31% identity with that of a nitrilotriacetate monooxygenase component. PMID- 10836750 TI - T(H)1 to T(H)2 shift of cytokines in peripheral blood of HIV-infected patients is detectable by reverse transcriptase polymerase chain reaction but not by enzyme linked immunosorbent assay under nonstimulated conditions. AB - BACKGROUND: Dysregulation of cytokines has been implicated in the pathogenesis of HIV infection. Therefore, we determined tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), IL-4, IL-10, and interferon-gamma (IFN-gamma) mRNA and serum levels in HIV-infected patients under nonstimulated conditions. MATERIAL AND METHODS: Blood samples of 32 HIV-infected patients and 10 healthy HIV-negative controls were analyzed. Cytokine serum levels were quantified by enzyme-linked immunosorbent assay (ELISA). Cytokine mRNA levels were determined semiquantitatively by competitive reverse transcriptase polymerase chain reaction (RT-PCR) and expressed as ratios relative to those of beta-actin. RESULTS: Competitive RT-PCR was shown to be more sensitive than protein ELISA in analyzing cytokine production. We found a significant correlation between steady-state mRNA ratios and serum protein levels for TNF-alpha. Significantly higher cytokine mRNA ratios were found in those patients with IL-10 and IFN-gamma levels detectable by ELISA. Steady-state mRNA ratios of TNF-alpha, IL-4, and IL-10 were significantly increased in patients with highly replicative HIV-infection. Furthermore, elevated IL-4:IFN-gamma ratios were related to both high viral load and loss of CD4 cells. DISCUSSION: Determination of steady-state mRNA ratios by semiquantitative RT-PCR represents a sensitive method to analyze cytokines in peripheral blood of HIV-infected patients under nonstimulated conditions. The data obtained with this technique provide further evidence for a T(H)1 to T(H)2 cytokine shift with progressive HIV disease. PMID- 10836751 TI - Depressive symptoms decline among persons on HIV protease inhibitors. AB - OBJECTIVE: To ascertain whether initiation of protease inhibitors was associated with a change in depressive symptoms among persons infected with HIV. METHODS: Study subjects included men and women who were enrolled in the HIV/AIDS Drug Treatment Program and who had completed an annual participant survey before and after initiating triple combination therapy with a protease inhibitor. Depressive symptoms were assessed using the Centre for Epidemiologic Studies-Depression scale (CES-D). Statistical analyses to determine the change in CES-D total and subscale scores before and after protease inhibitor use were conducted using parametric and multivariate methods. RESULTS: Our analysis was restricted to 453 participants. Of these 234 (52%) were depressed at baseline (CES-D score > or = 16). Compared with nondepressed participants, depressed participants were slightly younger (p = .048), less likely to be employed (p < .001) and more likely to have an annual income less than $10,000 per annum (p < .001). After adjusting for CD4 count, employment status, income, age, and CES-D total or subscale score at baseline, we found a significant improvement in total scale score (p = .001) and depressive mood (p = .002), positive affects (p = .005), and somatic symptoms (p = .011) subscale scores at follow-up. There was no significant change in the interpersonal relations score over the study period. CONCLUSION: Our findings indicate that in addition to conferring impressive clinical benefits, protease inhibitor use is associated with a significant improvement in HIV-positive individuals' mental health. PMID- 10836752 TI - Impact of zidovudine-based triple combination therapy on an AIDS drug assistance program. AB - A static deterministic model was used to estimate the effect of the shift to a triple combination therapeutic standard on the annual AIDS Drug Assistance Program (ADAP) budget, total medical care expenditures, and population health outcomes for New York (NY) state ADAP enrollees. The model used opportunistic disease incidence data from the Multicenter AIDS Cohort Study (MACS) and other studies. Costs of treating opportunistic infections (OIs) and other HIV complications with each type of therapy were derived from treatment algorithms and standard unit costs. CD4+ cell counts were used as an index of need for OI prophylaxis and for determining OI incidence. Treatment with zidovudine-based combination therapy has been shown to increase CD4+ cell counts and reduce OI incidence. The model estimated that a change from monotherapy to triple therapy would have increased NY ADAP budget expenditures per enrollee by 115%. However, total medical system costs per ADAP enrollee (including ADAP costs) would decrease by 0.4% in the base case as a result of reduction in OIs and other HIV sequelae and associated costs. Results are sensitive to the assumed percentage of people taking combination therapy as well as to the assumptions made about the impact of the combination therapy on CD4+ cell count. Total ADAP budget impacts will depend on the growth in ADAP enrollment as a result of the availability of more effective therapies. In conclusion, this model demonstrates how access to newer, more effective HIV drug treatments can reduce the costs of treating OIs and provide major health benefits for ADAP enrollees. PMID- 10836753 TI - Dissociation of immunologic and virologic responses to highly active antiretroviral therapy. AB - OBJECTIVE: Immunologic markers, levels of HIV DNA, and infectious HIV were compared in partial responders (PR) to HAART who had high plasma HIV RNA levels but stable or increasing levels of CD4+ peripheral blood mononuclear cells (PBMC), and patients with complete failure (CF) who had very low or decreasing levels of CD4+ PBMC and high plasma HIV RNA levels. DESIGN AND METHODS: CD4 and CD8 levels were monitored by flow cytometry. Beta2-microglobulin (beta2M) and neopterin levels were measured by quantitative enzyme immunoassays. Plasma and PBMC from 11 PR and 13 CF were analyzed for infectious HIV levels in limiting dilution cultures. Polymerase chain reaction (PCR) assays were used to quantify cellular HIV DNA and plasma HIV RNA. RESULTS: In comparison with CF, PR had little or no CD4+ cell loss, a substantial increase in CD8+ cells, significantly fewer positive plasma HIV cultures (p = .03), lower frequencies of infectious HIV in total PBMC (p = .005) and in CD4+ PBMC (p < .001), and lower frequencies of HIV DNA in CD4+ PBMC (p = .007). CONCLUSIONS: Lower levels of infectious HIV and a lower frequency of CD4+ PBMC that contain "productive" HIV DNA in PR as compared with CF may contribute to the stable or increasing CD4+ PBMC levels of the PR. However, HAART may also have effects on lymphocyte homeostasis independent of its antiviral activity. PMID- 10836754 TI - Improved antioxidant status among HIV-infected injecting drug users on potent antiretroviral therapy. AB - Low serum antioxidant levels in HIV-infected people have been attributed to altered metabolism associated with excess oxidative stress. We conducted a study to examine serum antioxidant levels in 175 HIV-positive and 210 HIV-negative injecting drug users (IDUs) in Baltimore, Maryland. At the time of data collection, 30 of the HIV-positive IDUs were receiving antiretroviral therapies (ART) including a protease inhibitor (PI), 43 ART without a PI, 22 monotherapies, and 80 not on any ART. Serum antioxidants examined included retinol, alpha tocopherol and gamma-tocopherol, alpha-carotene and beta-carotene, lycopene, lutein/zeaxanthin, and beta-cryptoxanthin. Mean serum levels of lycopene and lutein/zeaxanthin were significantly lower in HIV-positive IDUs than HIV-negative IDUs. Contrary to the findings in other studies, however, levels of the remaining antioxidants in HIV-positive study subjects were not lower than in HIV-negative study subjects. In fact, serum alpha-tocopherol levels were significantly higher in HIV-positive IDUs than HIV-negative IDUs (medians = 744 microg/dl and 718 microg/dl, respectively; p = .04). Among HIV-positive study subjects, there were significant differences in antioxidant levels by ART regimen. In multivariate models adjusting for injecting drug use, dietary intake, supplement intake, gender, and alcohol intake, significant overall differences by ART regimen were observed for alpha-tocopherol, beta-carotene, and beta-cryptoxanthin. Serum levels of these three antioxidants were significantly higher in the PI group than in the other three ART groups combined (p = .0008, 0.02, and 0.02, respectively). These data provide indirect evidence of the effectiveness of PIs in lowering oxidative stress levels in HIV-positive IDUs. PMID- 10836755 TI - The HIV epidemic in the Amazon Basin is driven by prototypic and recombinant HIV 1 subtypes B and F. AB - This paper describes genetic subtypes of HIV-1 found in blood samples from 31 HIV 1-infected people who visited the Counseling and Testing AIDS Center of Instituto de Medicina Tropical in Manaus, Brazil. Manaus, the main city in Brazil's Amazon Basin, is also the closest urban connection for more than 100,000 Indians living in the rain forests of this region. Although to date there is no evidence of increased incidence of HIV-1 infection among the indigenous population, our understanding of both the prevalence and nature of the epidemic in the region as a whole is limited. From the 31 samples analyzed by C2V3 sequencing, we found almost equal proportions of HIV-1 strains belonging to subtype B (n = 16; 51.6%) and subtype F (n = 15; 48.4%), a finding that differs from results from previous studies conducted in urban areas of southeastern Brazil. We also observed the presence of the GWGR amino-acid sequence in the critical tetra-peptide crown of the env V3 loop in the HIV-1 subtype B samples analyzed. Among these samples, we also found 14 mosaic genomes (45.16%) in which different combinations of subtypes B, C, and F were identified between the p24 gag, pro, and env regions. Our data support the hypothesis that the Amazonian HIV-1 infections linked to the urban epidemic in southeastern Brazil. The genetic diversity and the prevalence of mosaic genomes among the isolates in our study confirm an integral role of recombination in the complex Brazilian epidemic. PMID- 10836756 TI - Mortality of HIV-positive and HIV-negative heroin abusers as a function of duration of injecting drug use. AB - OBJECTIVE: To determine the incidence of mortality of injecting drug users as a function of the duration of injecting drugs and HIV status, and to assess how these effects vary according to age at initiation and calendar period (before and after 1992). METHODS AND DESIGN: Cohort of 376 intravenous heroin users admitted to detoxification between February 1987 and January 1990. SETTING: Patients referred from outpatient clinics of metropolitan Barcelona. Duration and characteristics of drug use were determined by interviews. Blood samples were collected during admission and analyzed for HIV, CD4+ cell count and different biologic parameters. Assessment of vital status and causes of death were obtained by hospital charts, death certificates, and autopsies. RESULTS: The study population consisted of 299 men and 77 women, whose mean age at entry was 26 years, mean duration of injecting drug use before admission 6.1 years; HIV seroprevalence at entry 70.2%. By the end of the follow-up (median 5.6 years), 21.8% of individuals had died (26.6% in HIV-positive, and 10.7% in HIV-negative injecting users). Based on Kaplan-Meier estimates, 10%, 20%, and 30% of HIV negative patients died by 8.7, 11.3 and 14.3 years, respectively, after initiating injecting drugs. The corresponding survival times for the seropositives were substantially lower: 6.6, 8.5, and 11.6 years, respectively. Overall, the survival time was significantly (p < .05) decreased by 22% in HIV positive injecting drug users. Older age at initiation of injecting drug use was significantly (p < .05) associated with mortality in HIV-positive heroin users but it showed the opposite direction among HIV-negative people. Death rates in HIV-positive patients of the same duration of drug use were similar in periods before and after 1992 (relative hazard (RH) = 0.97; 95% confidence interval: 0.58 1.61). Although not statistically significant, the hazard of death in HIV negative injecting drug users was substantially lower after 1992 (RH = 0.59). CONCLUSIONS: Before introduction of potent antiretroviral therapies, HIV infection further increased rates of mortality that had already been heightened by injecting drug use. Furthermore, HIV infection modifies the effect of age at initiation and eliminates the seemingly downward trend of mortality in HIV negative people. PMID- 10836757 TI - Repeat screening for HIV: when to test and why. AB - This paper presents models for repeat HIV screening under conditions of constant low HIV incidence. The models reveal a direct link between the prevalence of undetected HIV infection and the screening interval between repeat HIV tests. We show how to select screening intervals that either achieve a given HIV prevalence level, or optimally balance the cost of repeat HIV testing against the cost of HIV infection. Alternatively, given an existing repeat screening program, the model implies that cost of infection for which the given screening interval is optimal. The method also suggests how to select an HIV testing technology. The models are applied to existing repeat testing programs in the U.S. Army and among legal commercial sex workers in the state of Nevada in the Far West of the United States. PMID- 10836758 TI - Identifying human herpesvirus 8 infection: performance characteristics of serologic assays. AB - Epidemiologic studies of infection with the oncogenic human herpesvirus 8 (HHV-8) depend on serologic methods to diagnose infection. However, optimal strategies for identifying HHV-8 infection remain undefined. We therefore evaluated four enzyme-linked immunoassays (EIAs) and one immunofluorescence assay (IFA) using sera from 87 individuals with the prototype HHV-8 disease, Kaposi's sarcoma (KS), and 210 participants in a hemophilia study (who were presumed not to be infected with HHV-8). Assays performed reasonably well in distinguishing between infected and uninfected persons, with receiver operator curve areas between 0.86 and 0.96. Nonetheless, IFA had only 86% sensitivity and 88% specificity, and no EIA simultaneously had sensitivity and specificity above 90% for any of the optical density (OD) cutpoints used to define seropositivity. Some assays were markedly less sensitive with diluted KS sera, suggesting that they poorly identify low titer antibodies present in asymptomatic infection. We also developed a classification tree that categorized individuals as seropositive if they had OD > 2.00 on recombinant K8.1 protein EIA or if they had both K8.1 OD between 0.51 and 2.00 and positive IFA results; this strategy had between 80% and 90% sensitivity and 95% and 100% specificity. Overall, assays performed adequately for use in most epidemiologic investigations, but wider applications will require improved tests. PMID- 10836759 TI - Lipodystrophy: results of a data evaluation of patients receiving nelfinavir containing combination therapy. PMID- 10836760 TI - Hepatitis B virus seroconversion in HIV-HBV coinfected patients treated with highly active antiretroviral therapy. PMID- 10836761 TI - Dietary problems in African HIV-1-infected adults, Abidjan, Cote d'Ivoire. COTRIMO-CI Study Group. PMID- 10836762 TI - RNA viral load test for early diagnosis of vertical transmission of HIV-1 infection. PMID- 10836763 TI - Adherence to triple therapy and viral load response. PMID- 10836764 TI - Development of Kaposi sarcoma despite sustained suppression of HIV plasma viremia. PMID- 10836765 TI - How frequently should patients with type 2 diabetes mellitus be screened for retinopathy? PMID- 10836766 TI - How effective are antidepressant medications in the treatment of irritable bowel syndrome and nonulcer dyspepsia? PMID- 10836767 TI - Does losartan decrease all-cause mortality more than placebo or first-generation ACE inhibitors for patients with moderate to severe heart failure? PMID- 10836768 TI - What is the long-term prognosis for patients with Lyme disease? PMID- 10836769 TI - Does acute bronchitis really exist? A reconceptualization of acute viral respiratory infections. AB - BACKGROUND: Considerable overlap exists in patient presentations and physical findings in viral upper respiratory tract infections (URIs) and acute bronchitis. Our goal was to determine whether there are any clinical cues that could help physicians differentiate between these 2 conditions. METHODS: We performed a retrospective chart audit on 135 patients who had been given a diagnosis of acute bronchitis and a random sample of 409 patients with URIs over a 2.5-year period. Patient and provider characteristics, patient symptoms, and physical findings were compared with bivariate analyses and then entered into a logistic regression model. RESULTS: In bivariate analyses, a number of demographic variables, symptoms, and signs were associated with acute bronchitis. Multivariate analysis showed that the strongest independent predictors of acute bronchitis were cough (adjusted odds ratio [AOR]=21.12; 95% confidence interval [CI], 6.01-74.26), and wheezing on examination (AOR=12.16; 95% CI, 5.39-27.42). Nausea was the strongest independent predictor that the diagnosis would not be acute bronchitis (AOR=0.01; 95% CI, 0.01-0.85). However, there was considerable overlap between the 2 conditions, and the logistic model explained only 37% of the variation between the diagnoses. CONCLUSIONS: We hypothesize that sinusitis, URI, and acute bronchitis are all variations of the same clinical condition (acute respiratory infection) and should be conceptualized as a single clinical entity, with primary symptoms related to different anatomic areas rather than as different conditions. PMID- 10836770 TI - Predictors of antibiotic prescribing for nonspecific upper respiratory infections, acute bronchitis, and acute sinusitis. An UPRNet study. Upper Peninsula Research Network. AB - BACKGROUND: Antibiotics are often prescribed for viral respiratory infections. The goal of our study was to determine the factors associated with antibiotic prescribing for acute respiratory infections in primary care. METHODS: We performed an observational study in 15 primary care practices in Michigan using patient and physician surveys distributed during visits for acute respiratory infections. We included patients 4 years or older presenting with symptoms of an acute respiratory infection (n=482). The main outcome measures were prescriptions of antibiotics, signs and symptoms associated with antibiotic prescribing, and clinician-reported reasons for prescribing an antibiotic. RESULTS: We found that patients who were older than 18 years, sick for more than 14 days, and seen in urgent care clinics were more likely to receive antibiotics. Patients expected antibiotics if they perceived that the drug had helped with similar symptoms in the past. In an adjusted model, the variables significantly associated with antibiotic prescribing were physical findings of sinus tenderness (odds ratio [OR]=20.0; 95% confidence interval [CI], 8.3-43.2), rales/rhonchi (OR=19.9; 95% CI, 9.2-43.2), discolored nasal discharge (OR=11.7; 95% CI, 4.3-31.7), and postnasal drainage (OR=3.1; 95% CI, 1.6-6.0). The presence of clear nasal discharge on examination was negatively associated (OR=0.3; 95% CI, 0.2-0.5). CONCLUSIONS: Several physical signs play an important role in clinicians' decisions to prescribe antibiotics for respiratory infections. This information will be useful in designing interventions to decrease inappropriate antibiotic prescribing for upper respiratory infections. PMID- 10836771 TI - Prescribing antibiotics for upper respiratory infections. PMID- 10836772 TI - The effect of an illustrated pamphlet decision-aid on the use of prostate cancer screening tests. AB - BACKGROUND: Prostate cancer screening with serum prostate-specific antigen (PSA) and digital rectal examination (DRE) continues to increase. Our goal was to test the effect of a prostate cancer screening decision-aid on patients' knowledge, beliefs, and use of prostate cancer screening tests. METHODS: Our study was a randomized controlled trial of a prostate cancer screening decision-aid consisting of an illustrated pamphlet as opposed to a comparison intervention. We included 257 men aged 50 to 80 years who were receiving primary care at a Department of Veterans Affairs Hospital in Milwaukee, Wisconsin. The decision-aid provided quantitative outcomes of prostate cancer screening with DRE and PSA. We subsequently evaluated prostate cancer screening knowledge, beliefs, and test use. RESULTS: The illustrated pamphlet decision-aid was effective in improving knowledge of prostate cancer screening tests: 95% of the experimental group were aware of the possibility of false-negative test results compared with 85% of the comparison group (P <.01). Ninety-one percent of the experimental group were aware of the possibility of a false-positive screening test result compared with 65% of the comparison group (P <.01). However, there was no difference in the use of prostate cancer screening between the experimental (82%) and comparison (84%) groups, (P >.05). CONCLUSIONS: When used in a primary care setting, an illustrated pamphlet decision-aid was effective in increasing knowledge of prostate cancer screening tests but did not change the use of these tests. PMID- 10836773 TI - Decision-aids for prostate cancer screening. PMID- 10836774 TI - Changes in functional status related to health maintenance visits to family physicians. AB - BACKGROUND: Physicians do not provide preventive care at the level recommended by national organizations. This may be because of physicians' lack of training or low level of confidence or because of patients' fears, beliefs, and lack of health knowledge. METHODS: We used an observational prospective cohort study in an academic family practice office to investigate changes in patients' functional status associated with receiving recommendations to change behavior from family physicians. Patients 18 years and older presenting for health maintenance visits to family physicians completed a functional status instrument and a brief intake questionnaire by telephone before their visit. After the visit patients were randomized to a debriefing interview or an observation-only group. The interview included the Patient/Doctor Interaction Scale and an assessment of whether patients received a recommendation to change behavior. RESULTS: One hundred thirty-two patients were randomized to the debriefing group, and of those, 92% completed assessments at 3 months. Patients reporting recommendations to change behavior had lower scores at 1 and 3 months for mental health, social health, and self-esteem and higher anxiety and depression scores than patients not receiving these recommendations. CONCLUSIONS: There are declines in social and emotional functional status in patients presenting to family practice clinicians for health maintenance visits during which recommendations for behavioral change were made. Such declines may inhibit physicians from making recommendations for behavioral change or patients from accepting them. PMID- 10836775 TI - The effect of labeling on perceived ability to recover from acute illnesses and injuries. AB - BACKGROUND: The process of giving a patient a diagnosis may cause harm. The adverse effects of labeling, best documented for the diagnosis of hypertension, include increased absenteeism from work and lower earnings, increased depressive symptoms, and reduced quality of life. We tried to determine whether the diagnosis of hypertension affects perceptions about the time required to recover from common acute medical problems. METHODS: In an academic family practice clinic, equal numbers of patients with and without hypertension were asked to estimate how long it would take them to recover from an upper respiratory tract infection (URI), a urinary tract infection (UTI), and an ankle sprain now and 5 years ago (before the diagnosis of hypertension). RESULTS: Compared with patients who did not have hypertension, patients with hypertension estimated that it would take them twice as long, on average, to recover from a URI now (11.7 vs 6.0 days, P=.002) and in the past (10 vs 5.5 days, P=.02). These differences persisted after controlling for age, sex, race, and education. No significant differences were found for estimated recovery times for UTI or ankle sprain. CONCLUSIONS: The diagnosis of hypertension may affect patients' perceptions of their ability to recover from unrelated acute illnesses. This may have implications for the way physicians choose to present information to patients. PMID- 10836776 TI - Clinical preventive medicine: primum non nocere? PMID- 10836777 TI - The necessity of both anorectal and vaginal cultures for group B streptococcus screening during pregnancy. AB - BACKGROUND: Group B streptococcus (GBS) sepsis affects approximately 2 of every 1000 newborns. In an effort to decrease the incidence of neonatal GBS infection, the Centers for Disease Control and Prevention have established guidelines for screening and treatment during pregnancy. One strategy includes obtaining both vaginal and anorectal GBS cultures, then treating patients whose cultures are positive. Many of our patients are reluctant to undergo anorectal cultures. We conducted a study to determine whether performing cultures of both the vagina and anorectum would change patient management. METHODS: We obtained vaginal and anorectal GBS cultures from 222 consecutive patients at 35 to 37 weeks' gestation. RESULTS: Fifty-four patients (24.3%) had positive GBS cultures. Of those women, 10 (18.5%) had negative vaginal but positive rectal cultures. Thus, nearly one fifth of the patients with GBS colonization would not have received intrapartum antibiotics if only vaginal cultures had been performed. CONCLUSIONS: Health care providers caring for pregnant patients should consider obtaining both vaginal and anorectal cultures when screening for Group B streptococcus. PMID- 10836778 TI - The accuracy of community-based automated blood pressure machines. AB - BACKGROUND: Despite widespread use, the accuracy of community-based automated blood pressure machines has been questioned. We sought to determine if these machines are as accurate and reliable as those obtained by a clinician with a mercury manometer. METHODS: We randomly selected 25 pharmacies and compared blood pressure readings obtained from their automated machines with from a mercury manometer. We used 3 volunteers with arm circumferences at the low, medium, and high ends of the acceptable range of a normal adult cuff size. RESULTS: For the subject with the small arm size, store machines reported systolic pressure readings that were, on average, 10 mm Hg higher than those obtained by the clinician (P <.001) and diastolic pressures 9 mm Hg higher (P <.001). The mean systolic pressure readings for the subject with the medium arm size were not significantly different between the store machine and the mercury manometer, and the readings were only modestly different for diastolic pressure. For the subject with the large arm size, store machines reported diastolic pressure readings that were, on average, 8.3 mm Hg lower than those obtained using the mercury manometer (P <.001), but with no significant difference in the systolic pressure. CONCLUSIONS: We found that automated blood pressure machines from a representative community-based sample of pharmacies did not meet the accepted standards of accuracy and reliability. Accuracy of readings is especially uncertain for patients having arm sizes larger or smaller than average. PMID- 10836779 TI - Controlling blood glucose levels in patients with type 2 diabetes mellitus. An evidence-based policy statement by the American Academy of Family Physicians and American Diabetes Association. AB - OBJECTIVE: To review evidence about the benefit of intensive glycemic control for patients with type 2 diabetes and to develop practice recommendations. PARTICIPANTS: A 9-member panel composed of family physicians, general internists, endocrinologists, and a practice guidelines methodologist was assembled by the American Academy of Family Physicians, the American Diabetes Association, and the American College of Physicians. EVIDENCE: Admissible evidence included published randomized controlled trials and observational studies regarding the effects of glycemic control on microvascular and macrovascular complications and on adverse effects. We followed systematic search and data abstraction procedures. Greater weight was given to clinical trials and to evidence about health outcomes. CONSENSUS PROCESS: Interpretations of evidence and approval of documents were finalized by unanimous vote, with recommendations linked to evidence and not expert opinion. The full report was prepared by the chair and 2 panel members, representing each of the 3 organizations. The initial draft underwent external review by 14 diabetologists and family physicians and changes consistent with the evidence were incorporated. CONCLUSIONS: The evidence demonstrates that the risk of microvascular and neuropathic complications is reduced by lowering glucose concentrations. Whether glycemic control affects macrovascular outcomes is less clear. The potential benefits of glycemic control must be balanced against factors that either preempt benefits (eg, limited life expectancy, comorbid disease) or increase risk (eg, severe hypoglycemia). The magnitude of benefit is a function of individual clinical variables (eg, baseline glycated hemoglobin level, presence of preexisting microvascular disease). Appropriate targets for treatment should be determined by considering these factors, patients' risk profiles, and personal preferences. PMID- 10836780 TI - Strategies for changing clinicians' practice patterns. A new perspective. AB - How can we persuade clinicians to adopt proven practices? Education, incentives, feedback, social marketing, and various other change strategies have inconsistent and unpredictable effects. We propose a theoretical framework that can provide a reliable basis for selecting effective change strategies. We divide clinicians into 4 categories on the basis of their responses to new information about the effectiveness of clinical strategies. We similarly divide the universe of practice change strategies into knowledge-oriented and behavior-oriented methods. We then show why specific combinations of these strategies are likely to be consistently effective for each of the 4 categories of clinicians. PMID- 10836781 TI - Is amoxicillin more effective than placebo in treating acute otitis media in children younger than 2 years? PMID- 10836782 TI - Can a program of manual physical therapy and supervised exercise improve the symptoms of osteoarthritis of the knee? PMID- 10836783 TI - Is spiral (helical) computed tomography useful for diagnosing pulmonary embolism? PMID- 10836784 TI - What are the benefits and risks associated with the use of benzodiazepines to treat insomnia? PMID- 10836785 TI - Patients, interrupted? PMID- 10836786 TI - Functional equivalence of hairpins in the RNA subunits of RNase MRP and RNase P in Saccharomyces cerevisiae. AB - RNase MRP and RNase P are both ribonucleoprotein enzymes performing endonucleolytic cleavage of RNA. RNase MRP cleaves at a specific site in the precursor-rRNA transcript to initiate processing of the 5.8S rRNA. RNase P cleaves precursor tRNAs to create the 5' end of the mature tRNAs. In spite of their different specificities, the two RNases have significant structural similarities. For example, the two enzymes in Saccharomyces cerevisiae share eight protein subunits; only one protein is unique to each enzyme. The RNA components of the two nucleases also show striking secondary-structure similarity. To begin to characterize the role of the RNA subunits in enzyme function and substrate specificity, we swapped two hairpin structures (MRP3 and P3) between RNase MRP RNA and RNase P RNA of S. cerevisiae. The hairpins in the two enzymes could be exchanged without loss of function or specificity. On the other hand, when the MRP3 hairpin in RNase MRP of S. cerevisiae was replaced with the corresponding hairpin from the RNA of Schizosaccharomyces pombe or human RNase MRP, no functional enzyme was assembled. We propose that the MRP3 and P3 hairpins in S. cerevisiae perform similar functions and have coevolved to maintain common features that are different from those of MRP3 and P3 hairpins in other species. PMID- 10836787 TI - Molecular interactions and metal binding in the theophylline-binding core of an RNA aptamer. AB - An RNA aptamer containing a 15-nt binding site shows high affinity and specificity for the bronchodilator theophylline. A variety of base modifications or 2' deoxyribose substitutions in binding-site residues were tested for theophyllinebinding affinity and the results were compared with the previously determined three-dimensional structure of the RNA-theophylline complex. The RNA theophylline complex contains a U6-A28-U23 base triple, and disruption of this A28-U23 Hoogsteen-pair by a 7-deaza, 2'-deoxy A28 mutant reduces theophylline binding >45-fold at 25 degrees C. U24 is part of a U-turn in the core of the RNA, and disruption of this U-turn motif by a 2'-deoxy substitution of U24 also reduces theophylline binding by >90-fold. Several mutations outside the "conserved core" of the RNA aptamer showed reduced binding affinity, and these effects could be rationalized by comparison with the three-dimensional structure of the complex. Divalent ions are absolutely required for high-affinity theophylline binding. High-affinity binding was observed with 5 mM Mg2+, Mn2+, or Co2+ ions, whereas little or no significant binding was observed for other divalent or lanthanide ions. A metal-binding site in the core of the complex was revealed by paramagnetic Mn2+-induced broadening of specific RNA resonances in the NMR spectra. When caffeine is added to the aptamer in tenfold excess, the NMR spectra show no evidence for binding in the conserved core and instead the drug stacks on the terminal helix. The lack of interaction between caffeine and the theophylline-binding site emphasizes the extreme molecular discrimination of this RNA aptamer. PMID- 10836788 TI - Probing the structure of RNAIII, the Staphylococcus aureus agr regulatory RNA, and identification of the RNA domain involved in repression of protein A expression. AB - RNAIII, a 514-nt RNA molecule, regulates the expression of many Staphylococcus aureus genes encoding exoproteins and cell-wall-associated proteins. We have studied the structure of RNAIII in solution, using a combination of chemical and enzymatic probes. A model of the secondary structure was derived from experimental data with the help of computer simulation of RNA folding. The model contains 14 hairpin structures connected by unpaired nucleotides. The data also point to three helices formed by distant nucleotides that close off structural domains. This model was generally compatible with the results of in vivo probing experiments with dimethylsulfate in late exponential-phase cultures. Toe-printing experiments revealed that the ribosome binding site of hld, which is encoded by RNAIII, was accessible to the Escherichia coli 30S ribosomal subunit, suggesting that the in vitro structure represented a translatable form of RNAIII. We also found that, within the 3' end of RNAIII, the conserved hairpin 13 and the terminator form an intrinsic structural domain that exerts specific regulatory activity on protein A gene expression. PMID- 10836789 TI - Effects of anticodon 2'-O-methylations on tRNA codon recognition in an Escherichia coli cell-free translation. AB - The methylation of 2'-hydroxyl groups is one of the most common posttranscriptional modifications of naturally occurring stable RNA molecules. Some tRNA species have a 2'-O-methyl nucleoside at the first position of the anticodon, and it was suggested that this modification stabilizes the codon anticodon duplex. However, no tRNA species have been found to have the modification at the second or third position of the anticodon. In the present study, we measured the effects of anticodon 2'-O-methylation on the codon-reading efficiencies of the anticodon variants of the unmodified forms of Escherichia coli tRNA1(Ser), using a cell-free protein synthesis assay. The modification of C in the first position of the anticodon into 2'-O-methylcytidine increased the efficiency of reading the G-ending codon. On the other hand, the modifications of the second and/or third positions were detrimental to the codon-reading activity. Thus, 2'-hydroxyl groups at the second and third positions of the anticodon may have some role in the translation reaction, and this may be the reason why 2'-O methyl nucleosides are not found in these positions within natural tRNA species. PMID- 10836790 TI - In vitro analysis of the binding of ADAR2 to the pre-mRNA encoding the GluR-B R/G site. AB - The ADAR family of RNA-editing enzymes deaminates adenosines within RNA that is completely or largely double stranded. In mammals, most of the characterized substrates encode receptors involved in neurotransmission, and these substrates are thought to be targeted by the mammalian enzymes ADAR1 and ADAR2. Although some ADAR substrates are deaminated very promiscuously, mammalian glutamate receptor B (gluR-B) pre-mRNA is deaminated at a few specific adenosines. Like most double-stranded RNA (dsRNA) binding proteins, ADARs bind to many different sequences, but few studies have directly measured and compared binding affinities. We have attempted to determine if ADAR deamination specificity occurs because the enzymes bind to targeted regions with higher affinities. To explore this question we studied binding of rat ADAR2 to a region of rat gluR-B pre-mRNA that contains the R/G editing site, and compared a wild-type molecule with one containing mutations that decreased R/G site editing. Although binding affinity to the two sequences was almost identical, footprinting studies indicate ADAR2 binds to the wild-type RNA at a discrete region surrounding the editing site, whereas binding to the mutant appeared nonspecific. PMID- 10836791 TI - CCA initiation boxes without unique promoter elements support in vitro transcription by three viral RNA-dependent RNA polymerases. AB - It has previously been observed that the only specific requirement for transcriptional initiation on viral RNA in vitro by the RNA-dependent RNA polymerase (RdRp) of turnip yellow mosaic virus is the CCA at the 3' end of the genome. We now compare the abilities of this RdRp, turnip crinkle virus RdRp, and Qbeta replicase, an enzyme capable of supporting the complete viral replication cycle in vitro, to transcribe RNA templates containing multiple CCA boxes but lacking specific viral sequences. Each enzyme is able to initiate transcription from several CCA boxes within these RNAs, and no special reaction conditions are required for these activities. The transcriptional yields produced from templates comprised of multiple CCA or CCCA repeats relative to templates derived from native viral RNA sequences vary between 2:1 and 0.1:1 for the different RdRps. Control of initiation by such redundant sequences presents a challenge to the specificity of viral transcription and replication. We identify 3'-preferential initiation and sensitivity to structural presentation as two specificity mechanisms that can limit initiation among potential CCA initiation sites. These two specificity mechanisms are used to different degrees by the three RdRps. The finding that three viral RdRps representing two of the three supergroups within the positive-strand RNA viral RdRp phylogeny support substantial transcription in the absence of unique promoters suggests that this phenomenon may be common among positive-strand viruses. A framework is presented arguing that replication of viral RNA in the absence of unique promoter elements is feasible. PMID- 10836792 TI - A conserved hairpin structure in Alfamovirus and Bromovirus subgenomic promoters is required for efficient RNA synthesis in vitro. AB - The coat protein gene in RNA 3 of alfalfa mosaic virus (AMV; genus Alfamovirus, family Bromoviridae) is translated from the subgenomic RNA 4. Analysis of the subgenomic promoter (sgp) in minus-strand RNA 3 showed that a sequence of 37 nt upstream of the RNA 4 start site (nt +1) was sufficient for full sgp activity in an in vitro assay with the purified viral RNA-dependent RNA-polymerase (RdRp). The sequence of nt -6 to -29 could be folded into a potential hairpin structure with a loop represented by nt -16, -17, and -18, and a bulge involving nt -23. By introducing mutations that disrupted base pairing and compensatory mutations that restored base pairing, it was shown that base pairing in the top half of the putative stem (between the loop and bulge) was essential for sgp activity, whereas base pairing in the bottom half of the stem was less stringently required. Deletion of the bulged residue A-23 or mutation of this residue into a C strongly reduced sgp activity, but mutation of A-23 into U or G had little effect on sgp activity. Mutation of loop residues A-16 and A-17 affected sgp activity, whereas mutation of U-18 did not. Using RNA templates corresponding to the sgp of brome mosaic virus (BMV; genus Bromovirus, family Bromoviridae) and purified BMV RdRp, evidence was obtained indicating that also in BMV RNA a triloop hairpin structure is required for sgp activity. PMID- 10836793 TI - The location of protein S8 and surrounding elements of 16S rRNA in the 70S ribosome from combined use of directed hydroxyl radical probing and X-ray crystallography. AB - Ribosomal protein S8, which is essential for the assembly of the central domain of 16S rRNA, is one of the most thoroughly studied RNA-binding proteins. To map its surrounding RNA in the ribosome, we carried out directed hydroxyl radical probing of 16S rRNA using Fe(II) tethered to nine different positions on the surface of protein S8 in 70S ribosomes. Hydroxyl radical-induced cleavage was observed near the classical S8-binding site in the 620 stem, and flanking the other S8-footprinted regions of the central domain at the three-helix junction near position 650 and the 825 and 860 stems. In addition, cleavage near the 5' terminus of 16S rRNA, in the 300 region of its 5' domain, and in the 1070 region of its 3'-major domain provide information about the proximity to S8 of RNA elements not directly involved in its binding. These data, along with previous footprinting and crosslinking results, allowed positioning of protein S8 and its surrounding RNA elements in a 7.8-A map of the Thermus thermophilus 70S ribosome. The resulting model is in close agreement with the extensive body of data from previous studies using protein-protein and protein-RNA crosslinking, chemical and enzymatic footprinting, and genetics. PMID- 10836794 TI - Mtt1 is a Upf1-like helicase that interacts with the translation termination factors and whose overexpression can modulate termination efficiency. AB - Translation termination is the final step that completes the synthesis of a polypeptide. Premature translation termination by introduction of a nonsense mutation leads to the synthesis of a truncated protein. We report the identification and characterization of the product of the MTT1 gene, a helicase belonging to the Upfl-like family of helicases that is involved in modulating translation termination. MTT1 is homologous to UPF1, a factor previously shown to function in both mRNA turnover and translation termination. Overexpression of MTT1 induced a nonsense suppression phenotype in a wild-type yeast strain. Nonsense suppression is apparently not due to induction of [PSI+], even though cooverexpression of HSP104 alleviated the nonsense suppression phenotype observed in cells overexpressing MTT1, suggesting a more direct role of Hsp104p in the translation termination process. The MTT1 gene product was shown to interact with translation termination factors and is localized to polysomes. Taken together, these results indicate that at least two members of a family of RNA helicases modulate translation termination efficiency in cells. PMID- 10836795 TI - Puromycin-rRNA interaction sites at the peptidyl transferase center. AB - The binding site of puromycin was probed chemically in the peptidyl-transferase center of ribosomes from Escherichia coli and of puromycin-hypersensitive ribosomes from the archaeon Haloferax gibbonsii. Several nucleotides of the 23S rRNAs showed altered chemical reactivities in the presence of puromycin. They include A2439, G2505, and G2553 for E. coli, and G2058, A2503, G2505, and G2553 for Hf. gibbonsii (using the E. coli numbering system). Reproducible enhanced reactivities were also observed at A508 and A1579 within domains I and III, respectively, of E. coli 23S rRNA. In further experiments, puromycin was shown to produce a major reduction in the UV-induced crosslinking of deacylated (2N3A76)tRNA to U2506 within the P' site of E. coli ribosomes. Moreover, it strongly stimulated the putative UV-induced crosslink between a streptogramin B drug and m2A2503/psi2504 at an adjacent site in E. coli 23S rRNA. These data strongly support the concept that puromycin, along with other peptidyl transferase antibiotics, in particular the streptogramin B drugs, bind to an RNA structural motif that contains several conserved and accessible base moieties of the peptidyl transferase loop region. This streptogramin motif is also likely to provide binding sites for the 3' termini of the acceptor and donor tRNAs. In contrast, the effects at A508 and A1579, which are located at the exit site of the peptide channel, are likely to be caused by a structural effect transmitted along the peptide channel. PMID- 10836796 TI - A third member of the RNA-specific adenosine deaminase gene family, ADAR3, contains both single- and double-stranded RNA binding domains. AB - Members of the double-stranded RNA- (dsRNA) specific adenosine deaminase gene family convert adenosine residues into inosines in dsRNA and are involved in A-to I RNA editing of transcripts of glutamate receptor (GluR) subunits and serotonin receptor subtype 2C (5-HT(2C)R). We have isolated hADAR3, the third member of this class of human enzyme and investigated its editing site selectivity using in vitro RNA editing assay systems. As originally reported for rat ADAR3 or RED2, purified ADAR3 proteins could not edit GluR-B RNA at the "Q/R" site, the "R/G" site, and the intronic "hot spot" site. In addition, ADAR3 did not edit any of five sites discovered recently within the intracellular loop II region of 5 HT(2C)R RNAs, confirming its total lack of editing activity for currently known substrate RNAs. Filter-binding analyses revealed that ADAR3 is capable of binding not only to dsRNA but also to single-stranded RNA (ssRNA). Deletion mutagenesis identified a region rich in arginine residues located in the N-terminus that is responsible for binding of ADAR3 to ssRNA. The presence of this ssRNA-binding domain as well as its expression in restricted brain regions and postmitotic neurons make ADAR3 distinct from the other two ADAR gene family members, editing competent ADAR1 and ADAR2. ADAR3 inhibited in vitro the activities of RNA editing enzymes of the ADAR gene family, raising the possibility of a regulatory role in RNA editing. PMID- 10836797 TI - Isolation and characterization of polyadenylation complexes assembled in vitro. AB - We developed a two-step purification of mammalian polyadenylation complexes assembled in vitro. Biotinylated pre-mRNAs containing viral or immunoglobulin poly(A) sites were incubated with nuclear extracts prepared from mouse myeloma cells under conditions permissive for in vitro cleavage and polyadenylation and the mixture was fractionated by gel filtration; complexes containing biotinylated pre-mRNA and bound proteins were affinity purified on avidin-agarose resin. Western analysis of known components of the polyadenylation complex demonstrated copurification of polyadenylation factors with poly(A) site-containing RNA but not with control RNA substrates containing either no polyadenylation signals or a point mutation of the AAUAAA polyadenylation signal. Polyadenylation complexes that were assembled on exogenous RNA eluted from the Sephacryl column in fractions consistent with their size range extending from 2 to 4 x 10(6) Mr. Complexes endogenous to the extract were of approximately the same apparent size, but more heterogeneous in distribution. This method can be used to study polyadenylation/cleavage complexes that may form upon a number of different RNA sequences, an important step towards defining which factors might differentially associate with specific RNAs. PMID- 10836798 TI - Expression of hTERT and hTR in cis reconstitutes and active human telomerase ribonucleoprotein. AB - Telomeres in eukaryotic cells are generally synthesized and maintained by the ribonucleoprotein (RNP) telomerase. This enzyme is composed of at least two subunits, the telomerase reverse transcriptase (TERT) and the telomerase RNA. Human telomerase activity can be reconstituted in vitro by the expression of the telomerase protein catalytic subunit (hTERT) in the presence of recombinant human telomerase RNA (hTR) in a rabbit reticulocyte lysate (RRL) system. The hTERT and hTR subunits are independently expressed in vivo, and little is known about the mechanism of their assembly. To facilitate recombinant telomerase RNP formation and reconstitution, we engineered a construct, termed hTERT-hTR cis, in which the 3' end of the hTERT coding sequence was extended by the addition of the sequence encoding hTR. Expression of the hTERT-hTR cis construct in vitro (in RRL) and in vivo (in the yeast Saccharomyces cerevisiae) produced hTERT-hTR transcripts of the predicted size. Active human telomerase was reconstituted by hTERT-hTR cis expression in both RRL and S. cerevisiae. Assembly of functional human telomerase by the bicistronic expression of the protein and RNA components may facilitate the overexpression and reconstitution of this enzyme in heterologous systems. PMID- 10836799 TI - Assessing the quality of life in patients with cancer. PMID- 10836800 TI - Bromide-induced pseudohyperchloridemia. PMID- 10836801 TI - [Variability of pollination and of pollen in Cypress]. PMID- 10836802 TI - Variations in the individual thick lamellar properties within osteons by nanodentation. PMID- 10836803 TI - Caesarean section controversy. A debate is needed on caesarean section rates in India. PMID- 10836804 TI - Caesarean section controversy. Brazilian obstetricians are pressured to perform caesarean sections. PMID- 10836805 TI - Caesarean section controversy. Women choose caesarean section. PMID- 10836806 TI - Caesarean section controversy. Elective caesarean can increase the risk to the fetus. PMID- 10836807 TI - Caesarean section controversy. Further research is needed on why rates of caesarean section are increasing. PMID- 10836808 TI - Caesarean section controversy. The caesarean culture of Brazil. PMID- 10836809 TI - Serious hazard of transfusion (SHOT) initiative. Autologous transfusion would avoid many of the problems of blood transfusion. PMID- 10836810 TI - Establishing probable cause in cerebral palsy. There are problems with the consensus statement. PMID- 10836811 TI - The implications of outlawing age discrimination. Elderly people have different needs. PMID- 10836812 TI - Evaluation of hospital at home scheme. Another study found that patients prefer home care to hospital care. PMID- 10836813 TI - Use of hypericum as antidepressant. Naturalistic studies are needed. PMID- 10836814 TI - Use of hypericum as antidepressant. Active substances must be identified. PMID- 10836815 TI - Use of hypericum as antidepressant. The herbalist will see you now. PMID- 10836816 TI - Use of hypericum as antidepressant. Use of placebo in depression trial was unethical. PMID- 10836817 TI - Use of hypericum as antidepressant. Safety in overdose needs to be established. PMID- 10836818 TI - Doctors and theologians. Doctors may stop believing in the NHS. PMID- 10836819 TI - Caring for marginalised people. Leadership and strategy are needed to support those who provide care. PMID- 10836820 TI - Doctors and complementary medicine. Are medical dinosaurs heading for extinction? PMID- 10836821 TI - Commentary: Reporting of adverse events is worth the effort. PMID- 10836822 TI - A good death. Research on dying is scanty. PMID- 10836823 TI - A good death. Care pathway in Wales aims to improve care of dying patients. PMID- 10836824 TI - A good death. Principles of palliative care are yet to be applied in acute hospitals. PMID- 10836825 TI - A good death. Quality of death can be measured outside hospices. PMID- 10836826 TI - Population, consumption, and entrapment. Improve access to contraception to curb population growth. PMID- 10836827 TI - Will eradication of Helicobacter pylori improve symptoms of non-ulcer dyspepsia? This meta-analysis is potentially misleading. PMID- 10836828 TI - Will eradication of Helicobacter pylori improve symptoms of non-ulcer dyspepsia? Meta-analysis included unreliable studies. PMID- 10836829 TI - Will eradication of Helicobacter pylori improve symptoms of non-ulcer dyspepsia? More studies should have been included in meta-analysis. PMID- 10836830 TI - Use of unlabelled and off licence drugs in children. A European paediatric rule is needed to protect children. PMID- 10836831 TI - Mini-Symposium on Temporomandibular Disorders. Toronto, May 3, 1999. Proceedings. PMID- 10836832 TI - Sub-regional workshop on pollutant release and transfer registers (PRTRs), 4-5 May 1999, Bratislava, Slovak Republic. PMID- 10836833 TI - Ultrasound guided percutaneous thrombin injection for the treatment of iatrogenic pseudoaneurysms. PMID- 10836834 TI - Mobile intracardiac calcinosis: risk of thromboembolism in patients with haemodialysed end stage renal disease. PMID- 10836835 TI - Instantaneous pressure-flow velocity relations of systemic venous return in patients with univentricular circulation. PMID- 10836836 TI - Surgery for aortic stenosis in severely symptomatic patients older than 80 years: experience in a single UK centre. PMID- 10836837 TI - Flying after heart surgery. PMID- 10836838 TI - Radial coronary angiography and stenting. PMID- 10836839 TI - Pre-excitation or post-excitation. PMID- 10836840 TI - On the management of scorpion stings. PMID- 10836841 TI - Dr Carey Coombs and his non-existent cardiac infarct. PMID- 10836843 TI - Reinventing medicine. PMID- 10836842 TI - [Combined right-sided hernia of the semilunar line and deviational pathology of the proximal colon]. PMID- 10836845 TI - [Rem Viktorovich Petrov (on the 70th birthday)]. PMID- 10836844 TI - Self-reported sleep patterns and daytime sleepiness in the neurologically healthy aged. PMID- 10836846 TI - The Molecular Chirality Awards 1999 presented to Professors Yoshio Okamoto (Nagoya University), Kazuhiko Saigo (University of Tokyo), and Masakatsu Shibasaki *University of Tokyo), Japan. PMID- 10836847 TI - Identification of three novel mRNA splice variants of GITR. PMID- 10836848 TI - Playing guess the ointment. PMID- 10836849 TI - Erythema multiforme associated with hepatitis B immunization. PMID- 10836850 TI - Morphoea presenting shortly after the onset of Schmidt's syndrome. PMID- 10836851 TI - [Differential gout diagnosis. 2: Calcium in cartilage and soft tissues]. PMID- 10836852 TI - [Ultrasound: case examples for routine basic diagnosis]. PMID- 10836853 TI - Fetal cardiotocography and acid-base status during cesarean section. PMID- 10836854 TI - Gastric cancer cell lines lack Fas ligand (FasL) expression but kill T cells via a FasL independent pathway. PMID- 10836855 TI - Vector manometry and LOS dynamics. PMID- 10836856 TI - Mycophenolate mofetil for Crohn's disease. PMID- 10836857 TI - What are the clinical limitations of wide-diameter (4 mm or greater) root-form endosseous implants? PMID- 10836858 TI - Treatment of verrucae plana with 15% glycolic acid. PMID- 10836859 TI - Eruptive pseudoangiomatosis in an adult. PMID- 10836860 TI - Oral lichen planus induced by interferon-alpha-N1 in a patient with hepatitis C. PMID- 10836861 TI - Declaration by The Austrian Society of Dermatology and Venereology. PMID- 10836862 TI - Chinese elderly patients' perceptions of their rehabilitation needs following a stroke. PMID- 10836863 TI - Pen pals: correspondence as a method for data generation in qualitative research. AB - The study aimed to understand the impact of chronic illness on the lives of midlife women and explore and share the ways in which women adapt to and/or tolerate chronic illness in their lives. In 1998, 80 women participated in a study in which data were generated by corresponding with the researcher. Guided by feminist principles of collaboration, reciprocity and disclosure, we created rich stories about what it is like to live with a chronic illness. In this paper we will discuss the first phase of this inquiry which utilized correspondence between the researcher and the women. The issues posed by the use of correspondence as an innovative data generation process will be analysed. Correspondence, at first glance, may appear to be a rather impersonal communication medium. However, we are committed to this method of data generation and believe we have unlocked the doors to a viable qualitative research process. The literature to guide this process is scarce so we are keen to share work in progress. We will describe the preparation phase in setting up the study; discuss some practical issues, share some of the researcher's experiences in generating narratives from dialogues and hear from the women themselves what they consider to be significant about this research process. PMID- 10836864 TI - Evaluation of lymph node virus burden in human immunodeficiency virus-infected patients receiving efavirenz-based protease inhibitor--sparing highly active antiretroviral therapy. AB - Although efavirenz-containing regimens effectively suppress plasma levels of human immunodeficiency virus (HIV) RNA, it is now clear that undetectable plasma viremia may not reflect a lack of viral replication. Because lymphoid tissue is an active site of HIV replication, the lymph node virus burden was analyzed in persons who received highly active antiretroviral therapy (HAART) containing either efavirenz or a protease inhibitor (PI). Testing with in situ hybridization revealed no detectable follicular dendritic cell-associated HIV RNA in either group, and only 2 of 8 persons in the efavirenz group and 1 of 4 in the PI group had detectable RNA in lymph node mononuclear cells (LNMC) when tested by use of nucleic acid sequencebased amplification. Low levels of replication-competent HIV were identified in both groups by use of quantitative coculture assays. There was no evidence of development of resistance to either regimen in virus isolated from LNMC. These data support the use of efavirenz as an alternative to a PI in initial HAART regimens. PMID- 10836865 TI - No evidence for the involvement of interleukin 2 or the immunoglobulin heavy chain gene cluster in determining genetic susceptibility to multiple sclerosis. PMID- 10836866 TI - Coma in a patient with Alzheimer's disease taking low dose trazodone and gingko biloba. PMID- 10836867 TI - Bickerstaff's brainstem encephalitis subsequent to Campylobacter jejuni enteritis. PMID- 10836868 TI - Myelopathy associated with Hashimoto's disease. PMID- 10836871 TI - From the Centers for Disease Control and Prevention. Public opinion about public health--United States, 1999. PMID- 10836870 TI - From the Centers for Disease Control and Prevention. Escherichia coli O111:H8 outbreak among teenage campers--Texas, 1999. PMID- 10836869 TI - From the Centers for Disease Control and Prevention. HIV-related tuberculosis in a transgender Network--Baltimore, Maryland, and New York City Area, 1998-2000. PMID- 10836873 TI - JAMA patient page. Panic disorder. PMID- 10836872 TI - From the Centers for Disease Control and Prevention. Corporate action to reduce air pollution--Atlanta, Georgia, 1998-1999. PMID- 10836874 TI - AT(1)-R gene polymorphism. PMID- 10836875 TI - Transfusions of polymerized bovine hemoglobin in a patient with severe autoimmune hemolytic anemia. PMID- 10836876 TI - Mass psychogenic illness attributed to toxic exposure at a high school. PMID- 10836877 TI - Mass psychogenic illness attributed to toxic exposure at a high school. PMID- 10836878 TI - Mass psychogenic illness attributed to toxic exposure at a high school. PMID- 10836879 TI - Mass psychogenic illness attributed to toxic exposure at a high school. PMID- 10836880 TI - Mass psychogenic illness attributed to toxic exposure at a high school. PMID- 10836882 TI - Blood pressure and death from coronary heart disease. PMID- 10836881 TI - Blood pressure and death from coronary heart disease. PMID- 10836883 TI - Blood pressure and death from coronary heart disease. PMID- 10836884 TI - Unstable angina pectoris. PMID- 10836885 TI - Unstable angina pectoris. PMID- 10836886 TI - Unstable angina pectoris. PMID- 10836887 TI - Unstable angina pectoris. PMID- 10836888 TI - Impatient inpatient care. PMID- 10836889 TI - Impatient inpatient care. PMID- 10836890 TI - Increased glomerular filtration rate in kidney-transplant recipients who take sildenafil. PMID- 10836891 TI - Sildenafil and ocular perfusion. PMID- 10836892 TI - 9th Annual Summer Neuropeptide Conference. Key West, Florida, USA. 27 June-1 July 1999. Abstracts. PMID- 10836893 TI - Clinical practice guideline: diagnosis and evaluation of the child with attention deficit/hyperactivity disorder. American Academy of Pediatrics. AB - This clinical practice guideline provides recommendations for the assessment and diagnosis of school-aged children with attention-deficit/hyperactivity disorder (ADHD). This guideline, the first of 2 sets of guidelines to provide recommendations on this condition, is intended for use by primary care clinicians working in primary care settings. The second set of guidelines will address the issue of treatment of children with ADHD. The Committee on Quality Improvement of the American Academy of Pediatrics selected a committee composed of pediatricians and other experts in the fields of neurology, psychology, child psychiatry, development, and education, as well as experts from epidemiology and pediatric practice. In addition, this panel consists of experts in education and family practice. The panel worked with Technical Resources International, Washington, DC, under the auspices of the Agency for Healthcare Research and Quality, to develop the evidence base of literature on this topic. The resulting evidence report was used to formulate recommendations for evaluation of the child with ADHD. Major issues contained within the guideline address child and family assessment; school assessment, including the use of various rating scales; and conditions seen frequently among children with ADHD. Information is also included on the use of current diagnostic coding strategies. The deliberations of the committee were informed by a systematic review of evidence about prevalence, coexisting conditions, and diagnostic tests. Committee decisions were made by consensus where definitive evidence was not available. The committee report underwent review by sections of the American Academy of Pediatrics and external organizations before approval by the Board of Directors. The guideline contains the following recommendations for diagnosis of ADHD: 1) in a child 6 to 12 years old who presents with inattention, hyperactivity, impulsivity, academic underachievement, or behavior problems, primary care clinicians should initiate an evaluation for ADHD; 2) the diagnosis of ADHD requires that a child meet Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition criteria; 3) the assessment of ADHD requires evidence directly obtained from parents or caregivers regarding the core symptoms of ADHD in various settings, the age of onset, duration of symptoms, and degree of functional impairment; 4) the assessment of ADHD requires evidence directly obtained from the classroom teacher (or other school professional) regarding the core symptoms of ADHD, duration of symptoms, degree of functional impairment, and associated conditions; 5) evaluation of the child with ADHD should include assessment for associated (coexisting) conditions; and 6) other diagnostic tests are not routinely indicated to establish the diagnosis of ADHD but may be used for the assessment of other coexisting conditions (eg, learning disabilities and mental retardation). This clinical practice guideline is not intended as a sole source of guidance in the evaluation of children with ADHD. Rather, it is designed to assist primary care clinicians by providing a framework for diagnostic decisionmaking. It is not intended to replace clinical judgment or to establish a protocol for all children with this condition and may not provide the only appropriate approach to this problem. PMID- 10836894 TI - Socioeconomic variation in asthma hospitalization: excess utilization or greater need? PMID- 10836895 TI - Risk factors for invasive pneumococcal disease in children: a population-based case-control study in North America. PMID- 10836896 TI - Intravenous immunoglobulins versus sepsis. PMID- 10836897 TI - Monkeybar injuries. PMID- 10836898 TI - Is the G985A allelic variant of medium-chain acyl-CoA dehydrogenase a risk factor for sudden infant death syndrome? A pooled analysis. PMID- 10836899 TI - Evidence-based medicine. PMID- 10836900 TI - Index of suspicion. Case 2. Hyperexplexia. PMID- 10836901 TI - Index of suspicion. Case 3. Lipoid pneumonia. PMID- 10836902 TI - Comment on: caution on the use of altered fractionation for nasopharyngeal carcinoma. PMID- 10836903 TI - Different dose escalation from plan normalization scheme. PMID- 10836905 TI - Treatment of scorpion envenoming should include both a potent specific antivenom and support of vital functions. PMID- 10836906 TI - Supplement to the AHA guidelines for the management of transient ischemic attacks. PMID- 10836907 TI - Polymerase chain reaction amplification of bacterial 16s rRNA in biopsy samples. PMID- 10836908 TI - Paravaginal repair. PMID- 10836909 TI - Contributions from the meeting on Pasteurella and pasteurellosis. Brisbane, Australia, February 3-5, 1999. PMID- 10836910 TI - 1999 ASHG Award for Excellence in Education. Some future directions in medical genetics. PMID- 10836911 TI - Ardeparin sodium for extended out-of-hospital prophylaxis against venous thromboembolism after total hip or knee replacement. A randomized, double-blind, placebo-controlled trial. AB - BACKGROUND: The optimal duration of prophylaxis against venous thromboembolism after total hip or knee replacement is uncertain. OBJECTIVE: To determine the efficacy and safety of extended out-of-hospital prophylaxis with low-molecular weight heparin (ardeparin sodium). DESIGN: Randomized, double-blind, placebo controlled trial. SETTING: 33 community, university, or university-affiliated hospitals. PATIENTS: 1195 adults who had elective total hip or knee replacement and completed 4 to 10 days of postoperative ardeparin prophylaxis. INTERVENTION: Daily subcutaneous ardeparin (100 anti-Xa IU/kg of body weight) or placebo from time of hospital discharge to 6 weeks after surgery. MEASUREMENTS: Symptomatic, objectively documented venous thromboembolism or death, along with major bleeding, from time of hospital discharge to 12 weeks after surgery. RESULTS: Patients who received ardeparin (n = 607) and those who received placebo (n = 588) did not differ significantly in the cumulative incidence of venous thromboembolism or death (9 cases [1.5%] compared with 12 cases [2.0%]; odds ratio, 0.7 [95% CI, 0.3 to 1.7]; P > 0.2; absolute difference, -0.56 percentage points [CI, -2.2 to 1.1 percentage points]) or major bleeding (2 cases [0.3%] compared with 3 cases [0.5%]). CONCLUSIONS: Among patients who had total knee or total hip replacement and received 4 to 10 days of postoperative ardeparin prophylaxis, the cumulative incidence of symptomatic venous thromboembolism or death after hospital discharge was not significantly reduced by extended out-of hospital ardeparin prophylaxis. Extended ardeparin use could provide a maximum 2.2-percentage point true reduction in such events. The benefit of extended ardeparin use is not clinically important for most patients. Future research should identify high-risk patients who would benefit most from extended prophylaxis. PMID- 10836912 TI - Prognostic value of treadmill exercise testing in elderly persons. AB - BACKGROUND: Recent exercise testing guidelines recognized a gap in knowledge about the prognostic value of treadmill exercise testing in elderly persons. OBJECTIVE: To test the hypothesis that treadmill exercise testing has equal prognostic value among elderly (> or =65 years of age) and younger (<65 years of age) persons and to examine the incremental value of this testing over clinical data. DESIGN: Inception cohort with a median follow-up of 6 years. SETTING: Olmsted County, Minnesota. PATIENTS: All elderly (n = 514) and younger (n = 2593) residents of Olmsted County who underwent treadmill exercise testing between 1987 and 1989. MEASUREMENTS: Overall mortality and cardiac events (cardiac death, nonfatal myocardial infarction, and congestive heart failure). RESULTS: Compared with younger patients, elderly patients had more comorbid conditions, achieved a lower workload (6.0 and 10.7 metabolic equivalents; P < 0.001), and had a greater likelihood of a positive exercise electrocardiogram (28% and 9%; P < 0.001). With median follow-up of 6 years, overall survival (63% and 92%; P < 0.001) and cardiac event-free survival (66% and 95%; P < 0.001) were worse among elderly persons than among younger persons. Workload was the only treadmill exercise testing variable associated with all-cause mortality in both age groups, and the strength of association was similar. Workload and angina with exercise testing were associated with cardiac events in both age groups, whereas a positive exercise electrocardiogram was associated with cardiac events only in younger persons (P < 0.05 for all comparisons). After adjustment for clinical variables, workload was the only additional treadmill exercise testing variable that was predictive of death (P < 0.001) and cardiac events (P < 0.05); the strength of the association was similar in both age groups. Each 1-metabolic equivalent increase in exercise capacity was associated with a 14% and 18% reduction in cardiac events among younger and elderly persons, respectively. CONCLUSIONS: In elderly persons, treadmill exercise testing provided prognostic information that is incremental to clinical data. After adjustment for clinical factors, work-load was the only treadmill exercise testing variable that was strongly associated with outcome, and its prognostic effect was of the same magnitude in elderly and younger persons. PMID- 10836913 TI - Recombinant human relaxin in the treatment of scleroderma. A randomized, double blind, placebo-controlled trial. AB - BACKGROUND: Relaxin is a pregnancy-related hormone that has tissue remodeling and antifibrotic effects. Systemic sclerosis (scleroderma) is characterized by fibrosis of the skin, vasculature, and internal organs. OBJECTIVE: To assess the efficacy, safety, and dose-response effect of recombinant human relaxin in patients with scleroderma. DESIGN: Multicenter, parallel-group, randomized, double-blind, placebo-controlled trial. SETTING: Academic referral centers. PATIENTS: 68 patients who had had stable, diffuse scleroderma (moderate to severe) for less than 5 years. INTERVENTION: Recombinant human relaxin, 25 or 100 microg/kg of body weight per day, or placebo administered by continuous subcutaneous infusion over 24 weeks. MEASUREMENTS: Modified Rodnan skin score was the primary efficacy measure. Secondary measurements were pulmonary function, the Health Assessment Questionnaire, and other measures of scleroderma that reflected fibrosis. RESULTS: Patients who received 25 microg/kg of recombinant human relaxin per day had significantly lower skin scores than those who received placebo (mean change, -3.6 at 4 weeks [P = 0.021], -7.5 at 12 weeks [P < 0.001], and -8.7 at 24 weeks [P = 0.040]). Similar trends were noted in other outcome measures, including forced vital capacity, measures of oral aperture and hand extension, functional status, and global assessment. Patients who received 100 microg/kg of relaxin per day did not differ from those who received placebo. Drug related adverse events included menometrorrhagia, reversible anemia, and complications of the subcutaneous drug administration system (site irritation and local infection). CONCLUSIONS: Twenty-four weeks of recombinant human relaxin, 25 microg/kg per day, is associated with reduced skin thickening, improved mobility, and improved function in patients with moderate to severe diffuse scleroderma. PMID- 10836914 TI - Lower-extremity edema associated with gene transfer of naked DNA encoding vascular endothelial growth factor. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) promotes angiogenesis and vascular permeability. The extent to which VEGF may cause tissue edema in humans has not been established. OBJECTIVE: To evaluate patients undergoing VEGF gene transfer for evidence of lower-extremity edema. DESIGN: Prospective consecutive case series. SETTING: Hospital outpatient clinic. PATIENTS: 62 patients with critical limb ischemia and 28 patients with claudication. INTERVENTION: Gene transfer of VEGF DNA. MEASUREMENTS: Semiquantitative analysis of lower-extremity edema. RESULTS: Lower-extremity edema was observed in 31 of 90 (34%) patients. Edema was less common in patients with claudication than in those with pain at rest (P = 0.016) or ischemic ulcers (P < 0.001), and it was less common in patients with pain at rest than in those with ischemic ulcers (P= 0.017). Treatment was typically limited to a brief course of oral diuretics. CONCLUSIONS: Vascular endothelial growth factor may enhance vascular permeability in humans. At the doses of plasmid DNA used in this study, lower-extremity edema responded to oral diuretic therapy and did not seem to be associated with serious sequelae. PMID- 10836915 TI - Trimethoprim-sulfamethoxazole compared with ciprofloxacin for treatment and prophylaxis of Isospora belli and Cyclospora cayetanensis infection in HIV infected patients. A randomized, controlled trial. AB - BACKGROUND: In developing countries, Isospora belli and Cyclospora cayetanensis frequently cause chronic diarrhea in HIV-infected patients. OBJECTIVE: To compare 1 week of trimethoprim-sulfamethoxazole treatment and 1 week of ciprofloxacin treatment in HIV-infected patients with chronic diarrhea caused by I. belli and C. cayetanensis. DESIGN: Randomized, controlled trial. SETTING: HIV clinic in Port-au-Prince, Haiti. PATIENTS: 42 HIV-infected patients with chronic diarrhea due to I. belli (n = 22) or C cayetanensis (n = 20). INTERVENTIONS: Patients were randomly assigned to receive oral trimethoprim-sulfamethoxazole (160 mg or 800 mg) or ciprofloxacin (500 mg) twice daily for 7 days. Patients who responded clinically and microbiologically received prophylaxis for 10 weeks (1 tablet orally, three times per week). MEASUREMENTS: Treatment success was measured by cessation of diarrhea and negative stool examination at day 7. Prophylaxis success was measured by recurrent disease rate. RESULTS: Diarrhea ceased in all 19 patients treated with trimethoprim-sulfamethoxazole. Eighteen of 19 patients had negative results on stool examination at day 7 (95%). Among the 23 patients who received ciprofloxacin, diarrhea ceased in 20 (87% [CI; 66% to 97%]) and 16 had negative results on stool examination at day 7 (70%). By survival analysis, diarrhea from isosporiasis and cyclosporiasis ceased more rapidly with trimethoprim-sulfamethoxazole than with ciprofloxacin. All patients receiving secondary prophylaxis with trimethoprim-sulfamethoxazole remained disease-free, and 15 of 16 patients receiving secondary prophylaxis with ciprofloxacin remained disease-free. CONCLUSIONS: A 1-week course of trimethoprim-sulfamethoxazole is effective in HIV-infected patients with cyclosporiasis or isosporiasis. Although ciprofloxacin is not as effective, it is acceptable for patients who cannot tolerate trimethoprim-sulfamethoxazole. PMID- 10836916 TI - Faculty perceptions of gender discrimination and sexual harassment in academic medicine. AB - BACKGROUND: Gender-based discrimination and sexual harassment are common in medical practice and may be even more prevalent in academic medicine. OBJECTIVE: To examine the prevalence of gender-based discrimination and sexual harassment among medical school faculty and the associations of gender-based discrimination with number of publications, career satisfaction, and perceptions of career advancement. DESIGN: A self-administered mailed questionnaire of U.S. medical school faculty that covered a broad range of topics relating to academic life. SETTING: 24 randomly selected medical schools in the contiguous United States. PARTICIPANTS: A random sample of 3332 full-time faculty, stratified by specialty, graduation cohort, and sex. MEASUREMENTS: Prevalence of self-reported experiences of discrimination and harassment, number of peer-reviewed publications, career satisfaction, and perception of career advancement. RESULTS: Female faculty were more than 2.5 times more likely than male faculty to perceive gender-based discrimination in the academic environment (P < 0.001). Among women, rates of reported discrimination ranged from 47% for the youngest faculty to 70% for the oldest faculty. Women who reported experiencing negative gender bias had similar productivity but lower career satisfaction scores than did other women (P< 0.001). About half of female faculty but few male faculty experienced some form of sexual harassment. These experiences were similarly prevalent across the institutions in the sample and in all regions of the United States. Female faculty who reported being sexually harassed perceived gender-specific bias in the academic environment more often than did other women (80% compared with 61 %) and more often reported experiencing gender bias in professional advancement (72% compared with 47%). Publications, career satisfaction, and professional confidence were not affected by sexual harassment, and self-assessed career advancement was only marginally lower for female faculty who had experienced sexual harassment (P = 0.06). CONCLUSION: Despite substantial increases in the number of female faculty, reports of gender-based discrimination and sexual harassment remain common. PMID- 10836917 TI - The alcohol hangover. AB - PURPOSE: To review the cause, pathophysiologic characteristics, cost, and treatment of alcohol-induced hangover. DATA SOURCES: A MEDLINE search of English language reports (1966 to 1999) and a manual search of bibliographies of relevant papers. STUDY SELECTION: Related experimental, clinical, and basic research studies. DATA EXTRACTION: Data in relevant articles were reviewed, and relevant clinical information was extracted. DATA SYNTHESIS: The alcohol hangover is characterized by headache, tremulousness, nausea, diarrhea, and fatigue combined with decreased occupational, cognitive, or visual-spatial skill performance. In the United States, related absenteeism and poor job performance cost $148 billion annually (average annual cost per working adult, $2000). Although hangover is associated with alcoholism, most of its cost is incurred by the light-to-moderate drinker. Patients with hangover may pose substantial risk to themselves and others despite having a normal blood alcohol level. Hangover may also be an independent risk factor for cardiac death. Symptoms of hangover seem to be caused by dehydration, hormonal alterations, dysregulated cytokine pathways, and toxic effects of alcohol. Physiologic characteristics include increased cardiac work with normal peripheral resistance, diffuse slowing on electroencephalography, and increased levels of antidiuretic hormone. Effective interventions include rehydration, prostaglandin inhibitors, and vitamin B6. Screening for hangover severity and frequency may help early detection of alcohol dependency and substantially improve quality of life. Recommended interventions include discussion of potential therapies and reminders of the possibility for cognitive and visual-spatial impairment. No evidence suggests that alleviation of hangover symptoms leads to further alcohol consumption, and the discomfort caused by such symptoms may do so. Therefore, treatment seems warranted. CONCLUSIONS: Hangover, a common disorder, has substantial morbidity and societal cost. Appropriate management may relieve symptoms in many patients. PMID- 10836918 TI - The efficacy of "distant healing": a systematic review of randomized trials. AB - PURPOSE: To conduct a systematic review of the available data on the efficacy of any form of "distant healing" (prayer, mental healing, Therapeutic Touch, or spiritual healing) as treatment for any medical condition. DATA SOURCES: Studies were identified by an electronic search of the MEDLINE, PsychLIT, EMBASE, CISCOM, and Cochrane Library databases from their inception to the end of 1999 and by contact with researchers in the field. STUDY SELECTION: Studies with the following features were included: random assignment, placebo or other adequate control, publication in peer-reviewed journals, clinical (rather than experimental) investigations, and use of human participants. DATA EXTRACTION: Two investigators independently extracted data on study design, sample size, type of intervention, type of control, direction of effect (supporting or refuting the hypothesis), and nature of the outcomes. DATA SYNTHESIS: A total of 23 trials involving 2774 patients met the inclusion criteria and were analyzed. Heterogeneity of the studies precluded a formal meta-analysis. Of the trials, 5 examined prayer as the distant healing intervention, 11 assessed noncontact Therapeutic Touch, and 7 examined other forms of distant healing. Of the 23 studies, 13 (57%) yielded statistically significant treatment effects, 9 showed no effect over control interventions, and 1 showed a negative effect. CONCLUSIONS: The methodologic limitations of several studies make it difficult to draw definitive conclusions about the efficacy of distant healing. However, given that approximately 57% of trials showed a positive treatment effect, the evidence thus far merits further study. PMID- 10836919 TI - The tragic events of April 1996. AB - In the early 19th century, Port Arthur, Tasmania, was the site of a notorious prison in a land at the end of the world. In 1996, Port Arthur was also the site of the worst mass murder in modern Australian history. A gunman with a semiautomatic weapon stepped into a tourist coffee shop and systematically shot dead 35 men, women, and children. Throughout Australia, an outpouring of grief, shame, and anger followed this tragic event and led quickly to more stringent gun control legislation. Several years later, Australians still remember the mass murder at Port Arthur with shame and horror as a personal affront, rather than simply a historical event. In the more violent society of the United States, many Americans perceive themselves as helpless victims or detached observers, rather than as persons who are responsible for promoting change. PMID- 10836920 TI - Prolonged prophylaxis after joint replacement: another step sideways? PMID- 10836921 TI - Emerging immunotherapies for non-Hodgkin lymphomas: the tortoise approaches the finish line. PMID- 10836922 TI - Acne. PMID- 10836923 TI - Blood cultures drawn from indwelling central venous catheters. PMID- 10836924 TI - Name-based surveillance for HIV-infected persons. PMID- 10836925 TI - Physical therapy and exercise in osteoarthritis of the knee. PMID- 10836926 TI - Optimal acid suppression therapy in critically ill patients. PMID- 10836927 TI - Hepatopulmonary syndrome in noncirrhotic portal hypertension. PMID- 10836928 TI - Marked peripheral edema associated with montelukast and prednisone. PMID- 10836929 TI - Control group selection is an important but neglected issue in studies of antibiotic resistance. PMID- 10836930 TI - Thrombocytopenia after iron dextran administration in a patient with severe iron deficiency anemia. PMID- 10836931 TI - The language of medication-taking. PMID- 10836933 TI - The Young Woman at the Rise of the 21st Century: Gynecological and Reproductive Issues in Health and Disease. Conference proceedings. Athens, Greece, November 18 21, 1998. PMID- 10836932 TI - Proceedings of Postgraduate Course on the Staging of Lung Cancer. Genoa, Italy, December 15-16, 1998. PMID- 10836934 TI - Partnering for quality. PMID- 10836935 TI - Saliva cannot destroy HIV-infected milk or semen cells. PMID- 10836936 TI - AORN's role in the Best Practice Network. PMID- 10836937 TI - Importance of the regulatory process and regulation. PMID- 10836938 TI - Exploring the scope for advocacy by paediatricians. PMID- 10836939 TI - Failure to thrive. PMID- 10836940 TI - Gastrostomy feeding in the disabled child. PMID- 10836941 TI - Growth monitoring. PMID- 10836942 TI - GFR estimation in paediatric oncology. PMID- 10836943 TI - Helicobacter pylori infection with iron deficiency anaemia and subnormal growth at puberty. PMID- 10836944 TI - Imaging in scoliosis. PMID- 10836945 TI - Liverpool Infirmary for Children. PMID- 10836946 TI - Meningitis and meningococcal septicaemia. PMID- 10836947 TI - Nut allergy in children. PMID- 10836948 TI - Opportunistic immunisation in schools may be more effective than immunisation in hospital. PMID- 10836949 TI - Paracetamol overdosage. PMID- 10836950 TI - Perthes' disease and blood manganese levels. PMID- 10836951 TI - School nursing. PMID- 10836952 TI - Trying to deliver aerosols to upset children is a thankless task. PMID- 10836953 TI - Viruses in febrile convulsions. PMID- 10836954 TI - Epidemiology of whiplash. PMID- 10836955 TI - Epidemiology of whiplash. PMID- 10836956 TI - Epidemiology of whiplash. PMID- 10836957 TI - Diagnostic evaluation of classification criteria for RA and reactive arthritis. PMID- 10836958 TI - Efficacy of intra-articular primatised anti-CD4 in resistant rheumatoid knees. PMID- 10836959 TI - CD36 and CD14 immunoreactivity of Reiter cells in inflammatory synovial fluids. PMID- 10836960 TI - Non-period leg pain in patients with familial Mediterranean fever. PMID- 10836961 TI - From the Centers for Disease Control and Prevention. Surveillance for West Nile virus in overwintering mosquitoes--New York, 2000. PMID- 10836962 TI - From the Centers for Disease Control and Prevention. Progress in development of immunization registries--United States, 1999. PMID- 10836963 TI - From the Centers for Disease Control and Prevention. Age-specific excess deaths associated with stroke among racial/ethnic minority populations--United States, 1997. PMID- 10836964 TI - From the Centers for Disease Control and Prevention. Adoption of perinatal group B streptococcal disease prevention recommendations by prenatal-care providers- Connecticut and Minnesota, 1998. PMID- 10836965 TI - From the Centers for Disease Control and Prevention. Measles outbreak- Netherlands, April 1999-January 2000. PMID- 10836966 TI - Protection for human subjects in medical research. PMID- 10836967 TI - Protection for human subjects in medical research. PMID- 10836968 TI - Protection for human subjects in medical research. PMID- 10836969 TI - How should physicians involve patients in medical decisions? PMID- 10836970 TI - JAMA Patient Page: Organ transplantation. PMID- 10836971 TI - Monarch butterflies (Danus plexippus L.) use a magnetic compass for naviation. PMID- 10836972 TI - A highly conserved kinase is an essential component for stress tolerance in yeast and plant cells. PMID- 10836973 TI - Structure of renal organic anion and cation transporters. AB - Here we review the structural and functional properties of organic anion transporters (OAT1, OAT2, OAT3) and organic cation transporters (OCTN1, OCTN2, OCT1, OCT2, OCT3), some of which are involved in renal proximal tubular organic anion and cation secretion. These transporters share a predicted 12-transmembrane domain (TMD) structure with a large extracellular loop between TMD1 and TMD2, carrying potential N-glycosylation sites. Conserved amino acid motifs revealed a relationship to the sugar transporter family within the major facilitator superfamily. Following heterologous expression, most OATs transported the model anion p-aminohippurate (PAH). OAT1, but not OAT2, exhibited PAH-alpha ketoglutarate exchange. OCT1-3 transported the model cations tetraethylammonium (TEA), N(1)-methylnicotinamide, and 1-methyl-4-phenylpyridinium. OCTNs exhibited transport of TEA and/or preferably the zwitterionic carnitine. Substrate substitution as well as cis-inhibition experiments demonstrated polyspecificity of the OATs, OCTs, and OCTN1. On the basis of comparison of the structurally closely related OATs and OCTs, it may be possible to delineate the binding sites for organic anions and cations in future experiments. PMID- 10836974 TI - Everything you wanted to know about the bladder epithelium but were afraid to ask. AB - The mammalian urinary bladder epithelium (urothelium) performs the important function of storing urine for extended periods, while maintaining the urine composition similar to that delivered by the kidneys. The urothelium possesses four properties to perform this function. First, it offers a minimum epithelial surface area-to-urine volume; this reduces the surface area for passive movement of substances between lumen and blood. Second, the passive permeability of the apical membrane and tight junctions is very low to electrolytes and nonelectrolytes. Third, the urothelium has a hormonally regulated sodium absorptive system; thus passive movement of sodium from blood to urine is countered by active sodium reabsorption. Last, the permeability properties of the apical membrane and tight junctions of the urothelium are not altered by most substances found in the urine or blood. The importance of the barrier function of the urothelium is illustrated by infectious cystitis. The loss of the barrier function results in the movement of urinary constituents into the lamina propria and underlying muscle layers, resulting in suprapubic and lower back pain and frequent, urgent, and painful voiding. PMID- 10836975 TI - Effect of intraperitoneal pressures on tissue water of the abdominal muscle. AB - A major factor that affects solute and water transport through tissue is the state of tissue hydration. The amount of interstitial water directly affects the transport coefficients for both diffusion and convection. To investigate the effect of simultaneous exposure of tissue to hydrostatic and osmotic pressures on the state of tissue hydration and the pattern of distribution of tissue water, we dialyzed rats with isotonic (290 mosmol/kg) or hypertonic (510 mosmol/kg) solution at intraperitoneal pressures (P(ip)) between 0 and 6 mmHg, and we infused isotopic markers intravenously and determined their equilibrium distribution volumes (V(D)) in the anterior abdominal muscle (AAM) by quantitative autoradiography. Total tissue water volume (theta(TW)) was determined from dry-to-wet weight ratios. theta(urea), the V(D) of [(14)C]urea, equals the sum of the extracellular water volume (theta(EC), V(D) of [(14)C]mannitol) and intracellular water volume (theta(IC) = theta(urea) - theta(EC)). If theta(if) = interstitial water volume and theta(IV) = vascular water volume (V(D) of (131)I-labeled IgG), then theta(EC) = theta(if) + theta(IV). AAM hydrostatic pressure profiles were measured by a micropipette/servo-null system and demonstrated that elevation of P(ip) above 3 mmHg significantly (P < 0.05) increases mean tissue pressure (P(T)) to the same level regardless of intraperitoneal osmolality. The increase in P(T) resulted in a nonlinear tissue expansion primarily in the interstitium regardless of osmolality. From 0 to 6 mmHg, theta(if) (in ml/g dry tissue) increased from 0.59 +/- 0.02 to 1.7 +/- 0.05 and to 1.5 +/- 0.05 after isotonic and hypertonic dialysis, respectively, whereas theta(IC) increased from 2.8 +/- 0.08 to 3.0 +/- 0.1 after isotonic dialysis and decreased to 2.6 +/- 0.1 after hypertonic dialysis. After dialysis at 6 mmHg with isotonic or hypertonic solutions, theta(IV) increased from 0.034 +/- 0.001 to 0. 049 +/- 0.001 and 0.042 +/- 0.002, respectively. theta(urea) during hypertonic dialysis at P(ip) between 0 and 6 mmHg increased in a nonlinear fashion (F = 26.3, P < 0.001), whereas theta(IC) invariably decreased (F = 11.1, P < 0.001) and theta(if) doubled from its control value at low P(ip). In conclusion, elevation of intraperitoneal hydrostatic pressure causes tissue expansion, primarily in interstitium, irrespective of osmolality of the bathing solution. Tissue hydrostatic pressure is therefore the primary determinant of tissue properties with respect to hydration, which in turn affects diffusive and convective transport. PMID- 10836976 TI - Coincident microvillar actin bundle disruption and perinuclear actin sequestration in anoxic proximal tubule. AB - The present studies investigated acute disruption of microvillar actin cytoskeleton and actin association with other cytoskeletal components in ATP depleted rabbit proximal tubular cells. Video-enhanced differential-interference contrast microscopy and confocal microscopy were used to follow the fate of F actin during the disruption of microvilli. Within individual cells, all microvilli collapsed simultaneously. Microvillar actin filaments underwent a parallel decrease in length. Using a sequential cytoskeletal extraction protocol and electron microscopy, we revealed in the present studies the coincident sequestration of a distinct, perinuclear pool of actin that was primarily absent in control cells. Actin sequestration progressed in a duration-dependent manner, occurring as early as 15 min of anoxia when cellular ATP dropped to <5% of control level. Phalloidin staining and depolymerization treatment showed the majority (>90%) of this sequestered actin to be F-actin. A microvillar actin bundling protein villin was also sequestered in the same perinuclear complex of anoxic proximal tubules. In conclusion, the present results demonstrate a coincident microvillar actin bundle disruption and the perinuclear sequestration of F-actin in ATP-depleted proximal tubular cells. PMID- 10836977 TI - Carbonic anhydrase IV is expressed in H(+)-secreting cells of rabbit kidney. AB - Carbonic anhydrase (CA) IV is a membrane-bound enzyme that catalyzes the dehydration of carbonic acid to CO(2) and water. Using peptides from each end of the deduced rabbit CA IV amino acid sequence, we generated a goat anti-rabbit CA IV antibody, which was used for immunoblotting and immunohistochemical analysis. CA IV was expressed in a variety of organs including spleen, heart, lung, skeletal muscle, colon, and kidney. Rabbit kidney CA IV had two N-glycosylation sites and was sialated, the apparent molecular mass increasing by at least 11 to approximately 45 kDa in the cortex. Medullary CA IV was much more heavily glycosylated than CA IV from cortex or any other organ, such modifications increasing the molecular mass by at least 20 kDa. CA IV was expressed on the apical and basolateral membranes of proximal tubules with expression levels on the order of S2 > S1 > S3 = 0. Because CA IV is believed to be anchored to the apical membrane by glycosylphosphatidylinositol, the presence of basolateral CA IV suggests an alternative mechanism. CA IV was localized on the apical membranes of outer medullary collecting duct cells of the inner stripe and inner medullary collecting duct cells, as well as on alpha-intercalated cells. However, CA IV was not expressed by beta-intercalated cells, glomeruli, distal tubule, or Henle's loop cells. Thus CA IV was expressed by H(+)-secreting cells of the rabbit kidney, suggesting an important role for CA IV in urinary acidification. PMID- 10836978 TI - Vascular endothelial growth factor is a survival factor for renal tubular epithelial cells. AB - Vascular endothelial growth factor (VEGF) acts primarily as an endothelial cell mitogen via the "endothelial cell-specific" receptors VEGFR-1 (flt-1) and VEGFR-2 (flk-1/KDR). Only a few nonendothelial cells have been shown to possess functional VEGF receptors. We therefore examined the rat renal tubular epithelial cell line NRK52-E. NRK52-E expressed VEGFR-1 and VEGFR-2 mRNA and protein by RT PCR, Northern blotting, Western blotting, immunofluorescence, and ligand binding. Serum-starved NRK52-E incubated with VEGF showed a significant increase in [(3)H]thymidine incorporation compared with control (2.3-fold at 1-10 ng/ml, P < 0. 05; 3.3-fold at 50-100 ng/ml, P < 0.01). VEGF also protected NRK52-E from hydrogen peroxide-induced apoptosis and necrosis compared with control (annexin-V FITC-positive cells, 39 vs. 54%; viable cells, 50. 5 vs. 39.7%). Immunohistochemical staining using a variety of antibodies showed expression of both VEGF receptors in normal rat renal tubules in vivo. Because VEGF induced a proliferative and an antiapoptotic response in renal tubular epithelial cells, these data suggest that VEGF may act as a survival factor for renal tubular epithelium in vivo. PMID- 10836979 TI - Potassium restriction downregulates ROMK expression in rat kidney. AB - The ROMK family of proteins has biophysical properties and distribution within the kidney similar to those of secretory potassium channels of the distal nephron. To study the regulation of ROMK during variations in dietary potassium, we measured the abundance of ROMK protein in rat kidney by immunoblotting. Neither 2 nor 5 days of a high-potassium diet had an effect on protein abundance in the cortex or medulla. Potassium deprivation (2 or 5 days) decreased ROMK protein content in both cortical and medullary fractions, to 51 and 40% of controls, respectively. To see whether the Na-K-2Cl cotransporter is similarly affected by potassium restriction, we analyzed immunoblots by using an antibody for the rat type 1 bumetanide-sensitive cotransporter (BSC-1). Like ROMK, BSC-1 protein content was found to decrease significantly in the renal medulla of potassium-deprived rats. In the thick ascending limb of Henle's loop, a decrease in ROMK and BSC-1 could result in decreased reabsorption of NaCl, a finding associated with hypokalemia. PMID- 10836980 TI - Decreased abundance of major Na(+) transporters in kidneys of rats with ischemia induced acute renal failure. AB - Ischemia-induced acute renal failure (ARF) is known to be associated with significant impairment of tubular Na reabsorption. We examined whether temporary bilateral renal ischemia (30, 40, or 60 min) and reperfusion (1-5 days) affect the abundance of several renal Na transporters and urinary Na excretion (U(Na)V) in rats. In rats with mild ARF (30 min), immunoblotting revealed that proximal tubule type 3 Na(+)/H(+) exchanger (NHE-3) and type II Na-P(i) cotransporter (NaPi-II) were significantly decreased to 28 +/- 6 and 14 +/- 6% of sham levels, respectively, at day 1. Moreover, Na(+)-K(+)-ATPase levels were also significantly decreased (51 +/- 11%), whereas there was no significant decrease in type 1 bumetanide-sensitive cotransporter (BSC-1) and thiazide-sensitive cotransporter (TSC) levels. Consistent with reduced Na transporter abundance, fractional urinary Na excretion (FE(Na)) was significantly increased in mild ARF (30 min) and U(Na)V was unchanged, despite a marked reduction in glomerular filtration rate. Na transporter levels and renal Na handling were normalized within 5 days. Severe ischemic injury (60 min) resulted in a marked decrease in the abundance of Na(+)-K(+)-ATPase, NHE-3, NaPi-II, BSC-1, and TSC at both days 1 and 5. Consistent with this, FE(Na) was significantly increased at days 1 and 5. Intravenous K-melanocyte-stimulated hormone treatment partially prevented the ischemia-induced downregulation of renal Na transporters and reduced the high FE(Na) to control levels. We conclude that reduced levels of Na transporters along the nephron may play a critical role in the impairment of tubular Na reabsorption, and hence increased Na excretion, in ischemia-induced ARF. PMID- 10836981 TI - Upregulation of V(1) receptors in renal resistance vessels of rats developing genetic hypertension. AB - Previous studies have demonstrated that arginine vasopressin (AVP) produces exaggerated renal vasoconstriction in young spontaneously hypertensive rats (SHR) relative to normotensive rats. The exaggerated renal vascular reactivity does not appear to be due to a primary defect in postreceptor calcium signal transduction. Although the magnitudes of vascular responses differ, the relative proportions of calcium entry and mobilization pathways evoked by AVP in renal resistance vessels are similar in these rat strains. The purpose of the present study was to evaluate possible differences in V(1) mRNA and receptor density and affinity in preglomerular resistance vessels (<50 microm) obtained from young Wistar-Kyoto (WKY) and SHR. Quantitative RT-PCR analysis revealed twofold greater expression of the V(1a) receptor gene in preglomerular arterioles of 7-wk-old SHR compared with WKY. In vitro radiolabeled ligand binding studies were performed under equilibrium conditions on preglomerular resistance arterioles freshly isolated from kidneys of 7-wk-old rats. The results indicate that AVP receptor density (B(max)) is two to three times greater in SHR than in WKY (248 +/- 24 vs. 91 +/- 11 fmol/mg protein, P < 0.001). The affinity does not differ between strains (K(d) = 0.5 nM). Displacement studies yielded similar results for SHR and WKY. Unlabeled AVP completely displaced [(3)H]AVP binding, with an IC(50) of 2.5 x 10( 10) M. Expression of AVP receptor types in afferent arterioles was evaluated using the V(1) receptor agonist, [Phe(2), Ile(3),Org(8)]vasopressin, the V(1) receptor antagonist, [d(CH(2))(5), Tyr(Me)(2), Tyr(NH(2))(9)]Arg(8)-vasopressin, and the V(2) receptor agonist, desamino-[D-Arg(8)]vasopressin. Both the V(1) agonist and antagonist displaced up to 90% of the AVP binding with IC(50) values of 4 x 10(-8) and 8 x 10(-7) M, respectively. The V(2) receptor agonist was a weak inhibitor, displacing less than 15% of AVP binding at a high concentration of 10(-4) M. These results demonstrate that virtually all AVP receptors in the preglomerular arterioles are of the V(1) type. Collectively, our results provide evidence that the enhanced renal reactivity to AVP is mediated by a higher density of V(1) receptors associated with increased gene expression in renal resistance vessels of SHR developing genetic hypertension. PMID- 10836982 TI - Effects of 20-HETE and 19(S)-HETE on rabbit proximal straight tubule volume transport. AB - The kidney has the highest abundance of cytochrome P-450 of all extrahepatic organs. Within the kidney, the highest concentration of cytochrome P-450 is found in the proximal tubule. Whether 20- or 19(S)-hydroxyeicosatetraenoic acid (HETE), the major P-450 metabolites of arachidonic acid in the proximal tubule, affect transport in this segment has not been previously investigated. We examined the direct effects of 20- and 19(S)-HETE on volume absorption (J(v)) in the rabbit proximal straight tubule (PST). Production of 20-HETE by rabbit PST was demonstrated by incubating microdissected tubules with [(3)H]arachidonic acid and separating the lipid extract by HPLC. There was significant conversion of [(3)H]arachidonic acid to 20-HETE in control tubules that was inhibited by 10(-5) M N-methylsulfonyl-12,12-dibromododec-11-enamide (DDMS). Addition of exogenous 20 HETE had no effect on PST volume transport. However, inhibition of endogenous production of 20-HETE using DDMS stimulated transport. In the presence of DDMS, 20-HETE inhibited PST J(v). 19(S)-HETE in the bathing solution stimulated PST J(v) alone and in the presence of DDMS. Thus omega- and omega-1-hydroxylase products of arachidonic acid have direct effects on PST transport. Endogenous production of 20-HETE may play a role in tonic suppression of transport and may therefore be an endogenous regulator of transport in the proximal tubule. PMID- 10836983 TI - Store-operated Ca(2+) channels in human glomerular mesangial cells. AB - Experiments were performed to identify the biophysical properties of store operated Ca(2+) channels (SOC) in cultured human glomerular mesangial cells (MC). A fluorometric technique (fura 2) was utilized to monitor the change in intracellular calcium concentration ([Ca(2+)](i)) evoked by elevating external [Ca(2+)] from 10 nM to 1 mM (Delta[Ca(2+)]). Under control conditions, Delta[Ca(2+)] averaged 6 nM and was unaffected by elevating bath [K(+)]. After treatment with 1 microM thapsigargin to deplete the intracellular Ca(2+) store, the change in [Ca(2+)](i) (Delta[Ca(2+)](th)) averaged 147 +/- 16 nM. In thapsigargin-treated MC studied under depolarizing conditions (75 mM bath K(+)), Delta[Ca(2+)](th) was 45 +/- 7 nM. The Delta[Ca(2+)](th) response of thapsigargin treated cells was inhibited by La(3+) (IC(50) = 335 nM) but was unaffected by 5 microM Cd(2+). In patch clamp studies, inward currents were observed in cell attached patches with either 90 mM Ba(2+) or Ca(2+) in the pipette and 140 mM KCl in the bathing solution. The single-channel conductance was 2.1 pS with Ba(2+) and 0.7 pS with Ca(2+). The estimated selectivities were Ca(2+) > Ba(2+) >> K(+). These channels were sensitive to 2 microM La(3+), insensitive to 5 microM Cd(2+), and voltage independent, with an average channel activity (NP(o)) of 1.02 at command potential (-V(p)) ranging from 0 to -80 mV. In summary, MC exhibited an electrogenic Ca(2+) influx pathway that is suggestive of Ca(2+) entry through SOC, as well as a small-conductance divalent-selective channel displaying biophysical properties consistent with SOC. Based on estimates of whole cell Ca(2+) influx derived from our data, we conclude that SOC with low single-channel conductance must be highly abundant in MC to allow significant capacitative Ca(2+) entry in response to depletion of the intracellular store. PMID- 10836984 TI - Impaired myogenic autoregulation in kidneys of Brown Norway rats. AB - The Brown Norway (BN) rat is normotensive and has an extended lifespan but is extremely sensitive to hypertension-induced renal injury. Relative impairment of autoregulation has been implicated in the progression of renal failure whereas absence of myogenic autoregulation is associated with early renal failure. Therefore, we tested the hypothesis that there is conditional failure of renal autoregulation in BN rats. In isoflurane-anesthetized BN rats, the pressure-flow transfer function was normal when pressure fluctuated spontaneously. External forcing increased pressure fluctuation and exposed weakness of the myogenic component of autoregulation; the component mediated by tubuloglomerular feedback was less affected. In the presence of vasopressin to raise renal perfusion pressure, myogenic autoregulation was further impaired during forcing in BN rats but not in Wistar rats. Compensation by the myogenic system was rapidly restored on cessation of forcing, suggesting a functional limitation rather than a structural failure. Graded forcing in Wistar rats and in spontaneously hypertensive rats revealed that compensation due to the myogenic system was strong and independent of forcing amplitude. In contrast, graded forcing in BN rats showed that compensation was reduced when fluctuation of blood pressure was increased but that the reduction was independent of forcing amplitude. The results demonstrate conditional failure of myogenic autoregulation in BN rats. These acute studies provide a possible explanation for the observed sensitivity to hypertension-induced renal injury in BN rats. PMID- 10836985 TI - Specificity and functional analysis of the pH-responsive element within renal glutaminase mRNA. AB - The specificity and the functional significance of the binding of a specific cytosolic protein to a direct repeat of an eight-base AU sequence within the 3' nontranslated region of the glutaminase (GA) mRNA were characterized. Competition experiments established that the protein that binds to this sequence is not an AUUUA binding protein. When expressed in LLC-PK(1)-F(+) cells, the half-life of a beta-globin reporter construct, betaG-phosphoenolpyruvate carboxykinase, was only slightly affected (1.3-fold) by growth in acidic (pH 6.9, 10 mM HCO(-)(3)) vs. normal (pH 7.4, 25 mM HCO(-)(3)) medium. However, insertion of short segments of GA mRNA containing the direct repeat or a single eight-base AU sequence was sufficient to impart a fivefold pH-responsive stabilization to the chimeric mRNA. Furthermore, site-directed mutation of the direct repeat of the 8-base AU sequence in a betaG-GA mRNA, which contains 956 bases of the 3'-nontranslated region of the GA mRNA, completely abolished the pH-responsive stabilization of the wild-type betaG-GA mRNA. Thus either the direct repeat or a single eight-base AU sequence is both sufficient and necessary to create a functional pH-response element. PMID- 10836986 TI - Purification, characterization, and localization of an ATP diphosphohydrolase in porcine kidney. AB - Membranes of pig kidney cortex tissue were solubilized in the presence of Triton X-100. Partial purification of ATP diphosphohydrolase (ATPDase) was achieved by successive chromatography on concanavalin A-Sepharose, Q-Sepharose Fast Flow, and 5'-AMP-Sepharose 4B. Monoclonal antibodies against ATPDase were generated. Further purification of the ATPDase was obtained by immunoaffinity chromatography with these monoclonal antibodies. NH(2)-terminal amino acid sequencing of the 78 kDa protein showed a sequence very homologous to mammalian CD39. The protein is highly glycosylated, with a nominal molecular mass of approximately 57 kDa. The purified enzyme hydrolyzed di- and triphosphates of adenosine, guanosine, cytidine, uridine, inosine, and thymidine, but AMP and diadenosine polyphosphates could not serve as substrates. All enzyme activities were dependent on divalent cations and were partially inhibited by 10 mM sodium azide. The distribution of the enzyme in pig kidney cortex was examined immunohistochemically. The enzyme was found to be present in blood vessel walls of glomerular and peritubular capillaries. PMID- 10836987 TI - Adhesion of uric acid crystals to the surface of renal epithelial cells. AB - Adhesion of microcrystals that nucleate in tubular fluid to the apical surface of renal tubular cells could be a critical step in the formation of kidney stones, 12% of which contain uric acid (UA) either alone or admixed with calcium oxalates or calcium phosphates. UA crystals bind rapidly to monolayer cultures of monkey kidney epithelial cells (BSC-1 line), used to model the surface of the nephron, in a concentration-dependent manner. The urinary glycoproteins osteopontin, nephrocalcin, and Tamm-Horsfall glycoprotein had no effect on binding of UA crystals to the cell surface, whereas other polyanions including specific glycosaminoglycans blocked UA crystal adhesion. Specific polycations also inhibited adhesion of UA crystals and appeared to exert their inhibitory effect by coating cells. However, removal of anionic cell surface molecules with neuraminidase, heparitinase I, or chondroitinase ABC each increased UA crystal binding, and sialic acid-binding lectins had no effect. These observations suggest that hydrogen bonding and hydrophobic interactions play a major role in adhesion of electrostatically neutral UA crystals to renal cells, unlike the interaction of calcium-containing crystals with negatively charged molecules on the apical cell surface via ionic forces. After adhesion to the plasma membrane, subsequent cellular events could contribute to UA crystal retention in the kidney and the development of UA or mixed calcium and UA calculi. PMID- 10836988 TI - Amino acid transport in podocytes. AB - It has recently been shown that formation of podocyte foot processes is dependent on a constant source of lipids and proteins (Simons M, Saffrich R, Reiser J, and Mundel P. J Am Soc Nephrol 10: 1633-1639, 1999). Here we characterize amino acid transport mechanisms in differentiated cultured podocytes and investigate whether it may be disturbed during podocyte injury. RT-PCR studies detected mRNA for transporters of neutral amino acids (ASCT1, ASCT2, and B(0/+)), cationic AA (CAT1 and CAT3), and anionic AA (EAAT2 and EAAT3). Alanine (Ala), asparagine, cysteine (Cys), glutamine (Gln), glycine (Gly), leucine (Leu), methionine (Met), phenylalanine (Phe), proline (Pro), serine (Ser), threonine (Thr), glutamic acid (Glu), arginine (Arg), and histidine (His) depolarized podocytes and increased their whole cell conductances. Depletion of extracellular Na(+) completely inhibited the depolarization induced by Ala, Gln, Glu, Gly, Leu, and Pro and decreased the depolarization induced by Arg and His, indicating the presence of Na(+)-dependent amino acid transport. Incubation of podocytes with 100 microg/ml puromycin aminonucleoside for 24 h significantly attenuated the effects induced by the various amino acids by approximately 70%. The data indicate the existence of different amino acid transporter systems in podocytes. Alteration of amino acid transport may participate in podocyte injury and disturbed foot process formation. PMID- 10836989 TI - Bidirectional regulation of tonicity-responsive enhancer binding protein in response to changes in tonicity. AB - Tonicity-responsive enhancer binding protein (TonEBP) regulates transcription of tonicity responsive genes such as the sodium-myo-inositol cotransporter (SMIT), the sodium-chloride-betaine cotransporter (BGT1), and aldose reductase (AR). To characterize signals that activate TonEBP in Madin-Darby canine kidney (MDCK) cells, the abundance and nuclear distribution of TonEBP were studied after the osmolality of the culture medium was changed. Hypertonicity but not hyperosmolality is effective in activation of TonEBP as expected. Surprisingly, exposure to hypotonic medium leads to a dramatic downregulation of TonEBP both in abundance and nuclear distribution, indicating that under isotonic conditions, TonEBP is at a low-level activated state and can respond to both increase and decrease in tonicity. Additional experiments suggest that cellular ionic strength is the signal that initiates regulation of TonEBP. The increase in abundance of TonEBP is mediated by an increase in mRNA abundance and a parallel increase in synthesis of TonEBP. The stability of TonEBP mRNA is not affected by hypertonicity indicating that transcription plays a major role in the induction of TonEBP by hypertonicity. PMID- 10836990 TI - KCNA10: a novel ion channel functionally related to both voltage-gated potassium and CNG cation channels. AB - Our laboratory previously cloned a novel rabbit gene (Kcn1), expressed in kidney, heart, and aorta, and predicted to encode a protein with 58% amino acid identity with the K channel Shaker Kv1.3 (Yao X et al. Proc Natl Acad Sci USA 92: 11711 11715, 1995). Because Kcn1 did not express well (peak current in Xenopus laevis oocytes of 0.3 microA at +60 mV), the human homolog (KCNA10) was isolated, and its expression was optimized in oocytes. KCNA10 mediates voltage-gated K(+) currents that exhibit minimal steady-state inactivation. Ensemble currents of 5 10 microA at +40 mV were consistently recorded from injected oocytes. Channels are closed at the holding potential of -80 mV but are progressively activated by depolarizations more positive than -30 mV, with half-activation at +3.5 +/- 2.5 mV. The channel displays an unusual inhibitor profile because, in addition to being blocked by classical K channel blockers (barium tetraethylammonium and 4 aminopyridine), it is also sensitive to inhibitors of cyclic nucleotide-gated (CNG) cation channels (verapamil and pimozide). Tail-current analysis shows a reversal potential shift of 47 mV/decade change in K concentration, indicating a K-to-Na selectivity ratio of at least 15:1. The phorbol ester phorbol 12 myristate 13-acetate, an activator of protein kinase C, inhibited whole cell current by 42%. Analysis of single-channel currents reveals a conductance of approximately 11 pS. We conclude KCNA10 is a novel human voltage-gated K channel with features common to both K-selective and CNG cation channels. Given its distribution in renal blood vessels and heart, we speculate that KCNA10 may be involved in regulating the tone of renal vascular smooth muscle and may also participate in the cardiac action potential. PMID- 10836991 TI - Inhibition of type I and III IP(3)Rs by TGF-beta is associated with impaired calcium release in mesangial cells. AB - Inositol 1,4,5-trisphosphate receptors (IP(3)Rs) mediate cytosolic free calcium concentration ([Ca(2+)](c)) signals in response to a variety of agonists that stimulate mesangial cell contraction and proliferation. In the present study, we demonstrate that mesangial cells express both type I and III IP(3)Rs and that these receptors occupy different cellular locations. Chronic treatment with transforming growth factor-beta1 (TGF-beta1; 10 ng/ml, 24 h) leads to downregulation of both type I and III IP(3)Rs as measured by immunoblot and confocal analysis. TGF-beta1 treatment does not affect IP(3) levels, and downregulation of type I IP(3)R is not due to enhanced degradation of the protein, as the half-life of type I IP(3)R is unchanged in the presence or absence of TGF-beta1. Functional effects of TGF-beta1-induced downregulation of the IP(3)Rs were evaluated by measuring [Ca(2+)](c) changes in response to epidermal growth factor (EGF) in intact cells and sensitivity of [Ca(2+)](c) release to IP(3) in permeabilized cells. TGF-beta1 pretreatment led to a significant decrease of [Ca(2+)](c) release induced by EGF in intact cells and by submaximal IP(3) (400 nm) in permeabilized cells. Total IP(3)-sensitive [Ca(2+)](c) stores were not changed, as assessed by stimulation with maximal doses of IP(3) (10.5 microm) and thapsigargin-mediated calcium release in permeabilized cells. We conclude that prolonged exposure to TGF-beta1 leads to downregulation of both type I and III IP(3)Rs in mesangial cells and this is associated with impaired sensitivity to IP(3). PMID- 10836993 TI - Nucleic acid sequence and repair: role of adduct, neighbor bases and enzyme specificity. PMID- 10836992 TI - Luminal hypotonicity in proximal tubules of aquaporin-1-knockout mice. AB - To examine the role of aquaporin-1 (AQP1) in near-isosmolar fluid reabsorption in the proximal tubule, we compared osmolalities in micropuncture samples of late proximal tubular fluid and plasma in wild-type (+/+) and AQP1-knockout (-/-) mice. Compared with matched wild-type mice, the -/- animals produce a relatively hypotonic urine (607 +/- 42 vs. 1,856 +/- 101 mosmol/kgH(2)O) and have a higher plasma osmolality under micropuncture conditions (346 +/- 11 vs. 318 +/- 5 mosmol/kgH(2)O; P < 0.05). Measurements of tubular fluid osmolality were done in three groups of mice, +/+, -/-, and hydrated -/- mice in which plasma osmolality was reduced to 323 +/- 1 mosmol/kgH(2)O. Late proximal tubular fluid osmolalities were 309 +/- 5 (+/+, n = 21), 309 +/- 4 (-/-, n = 24), and 284 +/- 3 mosmol/kgH(2)O (hydrated -/-, n = 19). Tubular fluid chloride concentration averaged 152 +/- 1 (+/+), 154 +/- 1 (-/-), and 140 +/- 1 mM (hydrated -/-). Transtubular osmotic gradients in untreated and hydrated AQP1 -/- mice were 39 +/ 4 (n = 25) and 39 +/- 3 mosmol/kgH(2)O (n = 19), values significantly higher than in +/+ mice (12 +/- 2 mosmol/kgH(2)O; n = 24; both P < 0.001). AQP1 deficiency in mice generates marked luminal hypotonicity in proximal tubules, resulting from the retrieval of a hypertonic absorbate and indicating that near isosmolar fluid absorption requires functional AQP1. PMID- 10836994 TI - Hepatocyte growth factor upregulates E1AF that induces oral squamous cell carcinoma cell invasion by activating matrix metalloproteinase genes. AB - Hepatocyte growth factor (HGF) is thought to play a role in cell motility and invasion. Matrix metalloproteinases (MMPs) have been implicated in invasion and metastasis of tumor cells. We have previously reported that the Ets-oncogene family transcription factor E1AF positively regulates transcription of MMP genes in transient expression assays and that overexpression of the E1AF gene confers an invasive phenotype on breast cancer cells. Here we examined the effect of HGF on E1AF and MMP gene expression in terms of the invasive potential of the oral squamous cell carcinoma cell line HSC3. HGF stimulated expression of the E1AF gene. The levels of MMP-1, -3 and -9 mRNAs increased in cells treated with HGF and correlated with E1AF upregulation. In contrast, no obvious upregulation of MMP-1 and -9 mRNA was observed in ASE1AFHSC3 cells transfected with the antisense E1AF expression vector into parental HSC3 cells. The wild-type MMP-9 gene promoter was activated by endogenous E1AF in HSC3 cells, and chloramphenicol acetyltransferase (CAT) activities increased when HGF was added to transfected cells. On the other hand, CAT activity was reduced to almost two-thirds of the wild-type activity when HSC3 cells were transfected with a CAT reporter plasmid driven by a mutant MMP-9 promoter lacking the Ets-binding site, and induction of CAT activity was not observed upon addition of HGF. Analysis of organotypic raft cultures revealed that HSC3 cells invaded and degraded collagen gel actively upon addition of HGF. These results suggest that HGF induces expression of the Ets related E1AF transcription factor gene whose product in turn activates MMP genes and leads to oral cancer cell invasion. PMID- 10836995 TI - Apoptosis and cytokine release induced by ionizing or ultraviolet B radiation in primary and immortalized human keratinocytes. AB - We have compared the induction of apoptosis and cytokine release by UVB and gamma radiation in primary (untransformed) and in two immortalized human epithelial/keratinocyte cell lines, HaCaT and KB (KB is now known to be a subline of the ubiquitous keratin-forming tumour cell line HeLa and we therefore designate it HeLa-KB). In both the primary and the immortalized cell lines apoptosis and release of the inflammatory cytokine interleukin-6 are induced rapidly following UVB irradiation. In contrast, only the immortalized cells undergo apoptosis and release interleukin-6 after gamma-irradiation and here the onset of apoptosis and cytokine release are delayed. The same distinction between primary and immortalized cells was observed when double-strand breaks were induced with the anticancer drug mitoxantrone, which stabilizes topoisomerase II cleavable complexes. We suggest that immortalization may sensitize keratinocytes to the apoptogenic effect of ionizing radiation or mitoxantrone by deregulating normal cell cycle checkpoints. In both human keratinocytes and fibroblasts, cell killing, as assayed by loss of colony-forming ability, is not coupled to apoptosis. Immortalization increases resistance to gamma-radiation killing but sensitizes to apoptosis. In contrast, although immortalization also sensitizes to UVB-induced apoptosis, it does not affect UVB-induced cell killing. Apoptosis unambiguously indicates death at the single cell level but clonal cell survival integrates all the cellular and genetic processes which prevent or permit a scorable clone to develop. PMID- 10836996 TI - Reduced levels of connexin43 in cervical dysplasia: inducible expression in a cervical carcinoma cell line decreases neoplastic potential with implications for tumor progression. AB - Loss of gap junctional intercellular communication (GJIC) has been linked to aberrant proliferation and an enhanced neoplastic phenotype. Many human tumors, including the cervical carcinoma line HeLa, have been reported to be deficient in expression of the gap junction protein connexin43 (Cx43) and GJIC. To determine if this is an early event in carcinogenesis, we utilized immunohistochemistry to screen a series of cervical biopsy samples and demonstrated a major reduction in Cx43 expression in dysplastic regions compared to normal epithelia. To determine whether this loss influences the neoplastic behavior of cervical carcinoma cells, we have constructed HeLa cell lines in which Cx43 expression can be induced in response to doxycycline. This approach allows for the discrimination of Cx43 mediated effects from those due to pre-existing clonal heterogeneity. Cx43 induction in these cells led to assembly of functional junctions but did not alter growth control in vitro as measured by logarithmic growth, saturation density or focus formation when in co-culture with growth-controlled fibroblasts. However, Cx43 induction decreased two indices of neoplasia: it reduced anchorage independent growth and attenuated the growth rate of tumor xenografts. These results indicate that established HeLa cell lines are unresponsive to Cx43 mediated signals which are thought to mediate growth control of non-transformed cells, however, Cx43 expression can still reduce aspects of the neoplastic phenotype of these cells, indicating that loss of connexin signaling in dysplastic cells may contribute to their neoplastic progression. PMID- 10836997 TI - Helicobacter pylori causes DNA damage in gastric epithelial cells. AB - Helicobacter pylori infection has been considered as a risk factor for gastric carcinoma. Strong evidence exists that reactive oxygen species (ROS) play an important role in carcinogenesis, and in vivo investigations have shown increased synthesis of ROS in the gastric mucosa of H.pylori-infected patients. In the present study the direct effects of H.pylori on ROS and DNA synthesis, induction of apoptosis and DNA repair were investigated in the gastric epithelial cell lines AGS and HM02. Incubation of gastric cells with H.pylori extract induced the synthesis of ROS, diminished the levels of reduced glutathione (GSH), induced DNA fragmentation and increased DNA synthesis in gastric cells. Poly(ADP-ribose) formation was increased in gastric cells exposed to H.pylori extract. FACS analysis of gastric cells exposed to H.pylori extract did not reveal any change in the percentage of cells in the G(2)/M phase of the cell cycle. The radical scavengers MnTBAP (a cell permeable superoxide dismutase mimic), ebselen (a GSH peroxidase mimic) and high doses of catalase completely blocked H.pylori extract induced elevation in DNA synthesis. Our results indicate that H.pylori extract directly induces the synthesis of ROS in gastric epithelial cells and causes DNA damage. PMID- 10836998 TI - Frequent mutations of the beta-catenin gene in mouse colon tumors induced by azoxymethane. AB - The beta-catenin gene is frequently mutated at codons 33, 41 and 45 of the glycogen synthase kinase-3beta phosphorylation motif in human colon cancers in patients without APC mutations. Frequent mutations at codons 32 and 34, as well as 33 and 41, have been detected in rat colon tumors induced by azoxymethane (AOM), with the second G of CTGGA sequences being considered as a mutational hot spot. In the present study, exon 3 of the beta-catenin gene in mouse colon tumors induced by AOM was amplified by PCR and mutations were detected by the single strand conformation polymorphism method, restriction enzyme fragment length polymorphism and direct sequencing. All 10 colon tumors tested were found to have beta-catenin mutations, four in codon 34, three in codon 33, two in codon 41 and one in codon 37, nine being G:C-->A:T transitions. However, no mutations were found in codon 32 of the mouse beta-catenin gene. On immmunostaining, beta catenin was observed in the cytoplasm and nucleus of the tumor cells. The cytoplasmic staining was homogeneous, while both homogeneous and heterogeneous patterns were noted for the nuclei. Highly frequent mutations of the beta-catenin gene in AOM-induced mouse colon tumors suggest that consequent alterations in the stability and localization of the protein may play an important role in this colon carcinogenesis model. PMID- 10836999 TI - Biologic instability of pancreatic cancer xenografts in the nude mouse. AB - Tumor transplants into nude mice (NM) may reveal abnormal biological behavior compared with the original tumor. Despite this, human tumor xenografts in NM have been widely used to study the biology of tumors and to establish diagnostic and therapeutic modalities. Clearly, precise differences in the biology of a given tumor in human and in NM cannot be assessed. We compared the growth kinetics, differentiation pattern and karyotype of an anaplastic Syrian hamster pancreatic cancer cell line in NM and in allogenic hamsters. As with the original tumor, transplants in hamsters grew fast, were anaplastic and expressed markers related to tumor malignancy like galectin 3, TGF-alpha and its receptor EGFR at high levels. However, tumors in the NM were well-differentiated adenocarcinomas, grew slower, had increased apoptotic rate and had a high expression of differentiation markers such as blood group A antigen, DU-PAN-2, carbonic anhydrase II, TGF beta(2) and mucin. Karyotypically, the tumors in the NM acquired additional chromosomal damage. Our results demonstrate significant differences in the morphology and biology of tumors grown in NM and the allogenic host, and call for caution in extrapolating data obtained from xenografts to primary cancer. PMID- 10837000 TI - Diallyl disulfide inhibits p34(cdc2) kinase activity through changes in complex formation and phosphorylation. AB - Previous studies from our laboratory demonstrated that diallyl disulfide (DADS), an oil-soluble allyl sulfur compound found in processed garlic, markedly suppressed p34(cdc2) kinase activity and induced a G(2)/M phase arrest in cultured human colon tumor (HCT-15) cells. The present studies reveal that suppression of p34(cdc2) kinase activity by DADS does not result from direct interactions with the protein, but through changes in factors influencing the formation and conversion of the enzyme to its active form. Flow cytometric analyses showed that the increased proportion of cells in the G(2)/M phase following DADS treatment was accompanied by an increase in cyclin B(1) protein expression. A temporal and dose-dependent response in cyclin B(1) expression was observed in cells treated with DADS. Western blot analysis revealed that 50 microM DADS did not influence the quantity of p34(cdc2) protein expressed, but did decrease the amount associated with cyclin B(1) by 26% (P < 0.05). Exposure of unsynchronized cells to 25 or 50 microM DADS caused a trend towards increased p34(cdc2) hyperphosphorylation (17 and 22%, respectively). Exposure of synchronized cells to 100 microM DADS increased p34(cdc2) hyperphosphorylation by 15% (P < 0.05). Consistent with its ability to slightly increase the quantity of hyperphosphorylated p34(cdc2), DADS, 25 or 50 microM, decreased cdc25C protein expression by 23 and 46%, respectively (P < 0.05). The present studies suggest that the ability of DADS to inhibit p34(cdc2) kinase activation occurs because of decreased p34(cdc2)/cyclin B(1) complex formation and modest p34(cdc2) hyperphosphorylation. PMID- 10837001 TI - Comparative repair of the endogenous lesions 8-oxo-7, 8-dihydroguanine (8-oxoG), uracil and abasic site by mammalian cell extracts: 8-oxoG is poorly repaired by human cell extracts. AB - The repair of the endogenous lesions 8-oxo-7,8-dihydroguanine (8-oxoG), uracil (U) and natural abasic site (AP site) was investigated using an in vitro base excision repair assay in which a plasmid substrate containing a single lesion at a defined position was repaired by mammalian cell extracts. Repair replication of an 8-oxoG/cytosine base pair performed by normal human cell extracts was approximately 5-fold less efficient than repair of a U/adenine base pair and, in turn, the latter was repaired approximately 10-fold less efficiently than an AP site placed in front of an adenine. A similar pattern of repair capacity for the three lesions was observed in Chinese hamster extracts. Repair of 8-oxoG was performed by the one nucleotide insertion pathway only. The lower repair replication ability of 8-oxoG with respect to U was linked to a lower DNA glycosylase (base removal) activity rather than to inability to process the beta elimination cleaved strand left by the AP lyase activity associated with human oxoguanine DNA glycosylase 1. The data show that DNA repair of 8-oxoG is poor in human cells in comparison with other frequent endogenous lesions. PMID- 10837002 TI - 8-hydroxy-2'-deoxyguanosine in cervical cells: correlation with grade of dysplasia and human papillomavirus infection. AB - In this study, the 8-hydroxy-2'-deoxyguanosine (8-OHdG) level was assessed in human cervical cells by an immunoperoxidase method and was related to the presence of human papillomavirus (HPV) infection and precancerous lesions. After optimizing the immunohistochemical method of detecting oxidative DNA damage in whole cells, we have used this technique to estimate the oxidative damage in cervical cells collected during a routine PAP test. The analysis of variance (ANOVA) of the data from human samples showed significant differences in the 8 OHdG content among normal, low-grade and high-grade squamous intraepithelial lesion (SIL, HGSIL and LGSIL, respectively; P < 0.001). In the comparison of the three groups, statistically significant differences were detected between normal SIL and HGSIL (P < 0.001) and between LGSIL and HGSIL (P = 0.003), whereas no statistically significant difference was found between normal SIL and LGSIL (P = 0.1). Grouping observations by HPV status, no significant difference was detected in 8-OHdG levels between HPV(+) and HPV(-) subjects (P = 0.8). The polytomous and proportional odds models, extensions of the logistic regression analysis, showed that the effect of 8-OHdG levels in rising the risk of dysplasia was roughly constant through SIL grades. In conclusion, the immunoperoxidase method, applied to single human cervical cells, provides clear evidence that significant differences exist in 8-OHdG content between normal and dysplastic cells and that oxidative DNA damage might play an important role in cervical carcinogenesis. PMID- 10837003 TI - Efficacy of potential chemopreventive agents on rat colon aberrant crypt formation and progression. AB - We assessed the effects of 78 potential chemopreventive agents in the F344 rat using two assays in which the inhibition of carcinogen-induced aberrant crypt foci (ACF) in the colon was the measure of efficacy. In both assays ACF were induced by the carcinogen azoxymethane (AOM) in F344 rats by two sequential weekly injections at a dose of 15 mg/kg. Two weeks after the last AOM injection, animals were evaluated for the number of aberrant crypts detected in methylene blue stained whole mounts of rat colon. In the initiation phase protocol agents were given during the period of AOM administration, whereas in the post initiation assay the chemopreventive agent was introduced during the last 4 weeks of an 8 week assay, a time when ACF had progressed to multiple crypt clusters. The agents were derived from a priority listing based on reports of chemopreventive activity in the literature and/or efficacy data from in vitro models of carcinogenesis. During the initiation phase carboxyl amidoimidazole, p chlorphenylacetate, chlorpheniramine maleate, D609, diclofenac, etoperidone, eicosatetraynoic acid, farnesol, ferulic acid, lycopene, meclizine, methionine, phenylhexylisothiocyanate, phenylbutyrate, piroxicam, 9-cis-retinoic acid, S allylcysteine, taurine, tetracycline and verapamil were strong inhibitors of ACF. During the post-initiation phase aspirin, calcium glucarate, ketoprofen, piroxicam, 9-cis-retinoic acid, retinol and rutin inhibited the outgrowth of ACF into multiple crypt clusters. Based on these data, certain phytochemicals, antihistamines, non-steroidal anti-inflammatory drugs and retinoids show unique preclinical promise for chemoprevention of colon cancer, with the latter two drug classes particularly effective in the post-initiation phase of carcinogenesis. PMID- 10837004 TI - Brassica vegetables increase and apiaceous vegetables decrease cytochrome P450 1A2 activity in humans: changes in caffeine metabolite ratios in response to controlled vegetable diets. AB - Induction or inhibition of biotransformation enzymes, enzymes that activate or detoxify numerous xenobiotics, is one mechanism by which vegetables may alter cancer risk. Using a randomized crossover design, we examined the effect of various vegetable diets on cytochrome P450 (CYP) 1A2, N-acetyltransferase 2 (NAT2) and xanthine oxidase activity in humans. Men and women, non-smokers, on no medication and 20-40 years of age ate four 6-day controlled diets: basal (vegetable-free) and basal with three botanically defined vegetable groups. Enzyme activities were determined by measuring urinary caffeine metabolite ratios after a 200 mg caffeine dose on the last day of each feeding period. Mean CYP1A2 activity for 19 men and 17 women (least squares means adjusted for sex, GSTM1 genotype, urine volume and feeding period) with basal, brassica, allium and apiaceous vegetable diets differed significantly (P A nucleotide base shift negates the binding region for the general transcription factor SP1. Thus, individuals with the variant allele may be provided with a protective effect due to decreased metabolic conversion of carcinogenic compounds in tobacco smoke. This study has investigated the hypothesis that individuals with the variant allele may be at a reduced risk for lung cancer. Our results demonstrate that the protective effects of the MPO variant allele reduced overall lung cancer risk in Caucasians by 48% (OR = 0.52, 95% CI 0.30-0.90, P = 0.02). There was a 72% protective effect (OR = 0.28, 95% CI 0.12-0.61, P < 0.05) evident in men but not in women. Additionally, in younger individuals (<61 years) there was a statistically significant 72% reduction in risk (OR = 0.28, 95% CI 0. 11-0.69, P < 0.05) but not in older individuals. A protective effect was observed for current smokers (OR = 0.24, 95% CI 0.10-0.58, P < 0. 05) but not in former smokers and those who had never smoked. These data demonstrate that there is a reduction in lung cancer risk associated with a variant allele of MPO that is evident in men, younger individuals and current smokers. PMID- 10837006 TI - Beef induces and rye bran prevents the formation of intestinal polyps in Apc(Min) mice: relation to beta-catenin and PKC isozymes. AB - Epidemiological studies suggest that high consumption of red meat and saturated fat and low consumption of fiber are associated with an increased risk of colon cancer. Therefore, we studied whether diets high in red meat or high in different grain fibers as well as inulin, polydisperse beta(2-->1) fructan, could affect the formation of intestinal polyps in Apc(Min) mice. Min mice were fed the following high-fat (40% of energy) diets for 5-6 weeks; a high-beef diet and a casein-based diet without added fiber or casein-based diet with 10% (w/w) oat, rye or wheat bran, or 2.5% (w/w) inulin. One group had a normal low-fat AIN93-G diet. The mice fed the rye-bran diet had the lowest number of polyps in the distal small intestine [15.4 +/- 8.7 (mean +/- SD)], and in the entire intestine (26.4 +/- 12.1). The rye-bran group differed significantly (P = 0. 001-0.004) from the beef group (36.6 +/- 9.4 and 52.8 +/- 13.2). In addition, the beef group differed significantly from the AIN93-G group (P = 0.009) and also from the wheat bran group (21.0 +/- 6.1 and 35.0 +/- 8.2; P = 0.02) in the distal small intestine. The inulin group (32.9 +/- 14.3 and 49.3 +/- 16.3), on the other hand, was close to the beef group and it differed significantly from the rye-bran group in the distal small intestine. The number of animals bearing tumors in the colon + caecum was only 33% in the rye-bran group when compared with 89% in the beef and 100% in the inulin groups. The mice fed the rye-bran and beef diets had the lowest levels of cytosolic beta-catenin (0.60 +/- 0.42 and 0.67 +/- 0.26) and they differed significantly (P = 0.040 and 0.062) from the mice fed the oat-bran diet (1.46 +/- 0.43). No differences between groups in expression of protein kinase C (PKC) alpha, betaII, delta and zeta were found. The four PKC isozymes were positively correlated with cytosolic beta-catenin levels (r = 0.62-0.68; P < 0.0001). PMID- 10837007 TI - Role of glutathione in the accumulation of anticarcinogenic isothiocyanates and their glutathione conjugates by murine hepatoma cells. AB - Isothiocyanates (ITCs) are abundant in the human diet. Many potently inhibit tumorigenesis induced by a wide variety of chemical carcinogens in rodents. Recently, we observed that several ITCs accumulated to very high concentrations in cultured cells and that their accumulated levels were closely related to their potencies in inducing phase II enzymes [NAD(P)H:quinone reductase and glutathione transferases] that detoxify carcinogens. To elucidate the molecular mechanism responsible for this accumulation, the intracellular chemical identities of two ITCs, sulforaphane [SF, 1-isothiocyanato-(4R,S)-(methylsulfinyl)butane] and benzyl-ITC, were investigated in murine hepatoma cells. Both ITCs accumulated very rapidly to high intracellular concentrations, but, remarkably, most of the intracellular forms of the ITCs were dithiocarbamates resulting from conjugation with reduced glutathione (GSH). For example, the intracellular concentration reached 6.4 mM when cells were exposed to 100 microM SF for 30 min at 37 degrees C and 95% of the accumulated product was the GSH conjugate. Cellular accumulation of each ITC was accompanied by a profound reduction in cellular GSH levels. These findings, together with our previous observation that accumulation of ITCs depended on cellular GSH levels, strongly suggest that intracellular conjugation of ITCs with GSH is mainly responsible for ITC accumulation. Surprisingly, rapid accumulation to high concentrations also occurred when cells were exposed to the GSH-ITC conjugates. However, these conjugates were apparently not absorbed intact, but were hydrolyzed extracellularly to free ITCs that were taken up by the cells. This conclusion is supported by the finding that suppression of dissociation of the conjugates by excess GSH or other thiols blocks accumulation of the conjugates. PMID- 10837008 TI - Prevention of colonic aberrant crypt foci by dietary feeding of garcinol in male F344 rats. AB - The modifying effects of dietary feeding of a polyisoprenylated benzophenone, garcinol, isolated from Garcinia indica fruit rind on the development of azoxymethane (AOM)-induced colonic aberrant crypt foci (ACF) were investigated in male F344 rats. We also assessed the effects of garcinol on proliferating cell nuclear antigen (PCNA) index in ACF and activities of detoxifying enzymes of glutathione S-transferase (GST) and quinone reductase (QR) in liver. In addition, we examined the effects of garcinol on 12-O-tetradecanoylphorbol-13-acetate induced O(2)(-) generation in differentiated human promyelocytic HL-60 cells and lipopolysaccharide (LPS)- and interferon (IFN)-gamma-induced nitric oxide (NO) generation in mouse macrophage RAW 264.7 cells. Western blotting analysis of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression was done in LPS- and IFN-gamma-treated mouse macrophage RAW 264.7 cells. Rats were given subcutaneous injections of AOM (15 mg/kg body wt) once a week for 3 weeks to induce ACF. They also received the experimental diet containing 0.01 or 0.05% garcinol for 5 weeks, starting 1 week before the first dosing of AOM. AOM exposure produced 97 +/- 15 ACF/rat at the end of the study (week 5). Dietary administration of garcinol caused significant reduction in the frequency of ACF: 72 +/- 15 (26% reduction, P < 0.01) at a dose of 0.01% and 58 +/- 8 (40% reduction, P < 0.001) at a dose of 0.05%. Garcinol administration significantly lowered PCNA index in ACF. Feeding of garcinol significantly elevated liver GST and QR activities. In addition, garcinol could suppress O(2)(-) and NO generation and expression of iNOS and COX-2 proteins. These findings might suggest possible chemopreventive ability of garcinol, through induction of liver GST and QR, inhibition of O(2)(-) and NO generation and/or suppression of iNOS and COX-2 expression, on colon tumorigenesis. PMID- 10837009 TI - Detection of 1,N(2)-propanodeoxyguanosine adducts in DNA of Fischer 344 rats by an adapted (32)P-post-labeling technique after per os application of crotonaldehyde. AB - Crotonaldehyde is an important industrial chemical to which humans and animals are ubiquitously exposed. The main intake occurs via food, tobacco smoke and possibly also via beverages. Estimation of intake via the different routes is difficult since the data available on exposure are inconsistent. Crotonaldehyde is genotoxic, mutagenic and carcinogenic and forms 1,N(2)-propanodeoxyguanosine adducts as the main DNA adducts. We have developed a (32)P-post-labeling method for these adducts based on nuclease P1 enrichment and polyethyleneimine-cellulose TLC which allows reliable detection with a detection limit of 3 adducts/10(9) nucleotides, a labeling efficiency of 80-90% and a recovery of 38%. Using this method we found crotonaldehyde adducts in different organs of Fischer 344 rats after a single gavage of high doses of 300 and 200 mg/kg body wt in the range 0.3 3.2 +/- 0.4 adducts/10(8) nucleotides and after repeated gavage of low doses of 10 and 1 mg/kg body wt (five times a week for 6 weeks) 6.2 +/- 0.2 and 2.0 +/- 0.4 adducts/10(8)nucleotides, but not in untreated animals nor in calf thymus DNA not treated with crotonaldehyde. In contrast to our results, Chung and co-workers found adducts in tissue of untreated Fischer 344 rats. This discrepancy could depend on the different methods used but also on differences in exposure of the animals via food or due to animal housing, etc. PMID- 10837010 TI - N-acetyltransferase-dependent activation of 2-hydroxyamino-1-methyl-6 phenylimidazo[4,5-b]pyridine: formation of 2-amino-1-methyl-6-(5 hydroxy)phenylimidazo [4,5-b]pyridine, a possible biomarker for the reactive dose of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine. AB - 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) is a mutagenic and carcinogenic heterocyclic amine formed during ordinary cooking. PhIP is metabolically activated to the ultimate mutagenic metabolite by CYP P450-mediated N-hydroxylation followed by phase II esterification. Incubation of N-hydroxy-PhIP (N-OH-PhIP) with cytosol, acetyl coenzyme A (AcCoA) and 2'-deoxyguanosine for 24 h resulted in the formation of three different adducts:N(2)-(deoxyguanosin-8-yl) PhIP, N(2)-(guanosin-8-yl)-PhIP and PhIP-xanthine. One additional product, 5 hydroxy-PhIP (5-OH-PhIP), was also identified in the incubation mixtures. 5 hydroxy-PhIP is formed as a degradation product of conjugates formed from N acetoxy-PhIP and protein, glutathione or buffer constituents. A similar spectrum of products was obtained using 3'-phosphoadenosine-5'-phosphosulfate (PAPS) instead of acetyl CoA. Addition of glutathione (3 mM) to the incubation mixture resulted in a 50% reduction in both adducts and 5-hydroxy-PhIP formation in liver cytosol. The main product detected was PhIP, suggesting glutathione-dependent reduction of the N-acetoxy-PhIP. Addition of glutathione to incubation mixtures from the other cytosolic preparations had less dramatic effects. In addition, increasing the amount of N-OH-PhIP in the incubation mixture resulted in proportional increased amounts of total adducts and 5-OH-PhIP. Incubation of rat and human S9 with PhIP resulted in the formation of only traces of 5-OH-PhIP. Fortification with AcCoA clearly increased the formation of 5-OH-PhIP. Addition of the CYP 450 1A2 inhibitor, furafylline, completely inhibited the formation of 5-OH-PhIP in incubations with human S9. These results indicate that both PhIP adducts and 5-OH-PhIP are formed by similar routes of activation of N-OH-PhIP. 5 OH-PhIP may therefore serve as a biomarker for the formation of the ultimate mutagenic metabolite of PhIP. A rat dosed orally with PhIP excreted 1% of the dose as 5-OH-PhIP in the urine at 24 h and 0.05 and 0.01% at 48 and 72 h, respectively. This shows that 5-OH-PhIP is also formed in vivo and indicates the possible use of 5-OH-PhIP as a urinary biomarker. PMID- 10837011 TI - Inhibition of TGF-beta-induced apoptosis by ethinyl estradiol in cultured, precision cut rat liver slices and hepatocytes. AB - Ethinyl estradiol (EE) is a strong promoter of hepatocarcinogenesis in the rat. Treatment with EE and other hepatic promoters induces transient growth stimulation followed by growth inhibition (mitosuppression) in hepatocytes. Previously, we identified several genes whose transcript levels were increased during EE-induced mitosuppression, including transforming growth factor beta (TGF beta), which inhibits growth and induces apoptosis in hepatocytes. Various hepatic promoters, including phenobarbital and several peroxisomal proliferators, have been shown to inhibit TGF-beta-induced apoptosis in rat hepatocytes. The goal of this study was to investigate whether EE is also an inhibitor of TGF-beta induced apoptosis in rat hepatocytes. Several approaches to detect apoptosis were used, including the TUNEL assay, detection of high molecular weight DNA fragmentation by field inversion gel electrophoresis and determination of cytosolic cytochrome c levels by western analysis. TGF-beta-induced apoptosis in cultured, precision cut liver slices and hepatocytes of female rats. EE (95% in human liver microsomes, liver cytosol and red blood cells, respectively. In rat liver microsomes, (R)- and (S)-NNAL were metabolized at similar rates by alpha hydroxylation, considered to be the major metabolic activation pathway of NNAL. Pyridine-N-oxidation and adenosine dinucleotide phosphate adduct formation also occurred at similar rates from both enantiomers, while reoxidation to NNK was favored with (S)-NNAL as substrate. In rat lung microsomes, (S)-NNAL was more rapidly metabolized than (R)-NNAL by all oxidative pathways. In human liver microsomes, there were no significant differences in the rates of alpha hydroxylation, pyridine-N-oxidation and reoxidation to NNK between the two enantiomers. The results of this study demonstrate that (S)-NNAL, the more tumorigenic enantiomer in mice, is preferentially formed from NNK in rodent and human tissues, and is a substrate for oxidative metabolism in rodent and human tissue microsomes. PMID- 10837016 TI - Mutagen sensitivity and environmental exposures as contributing causes of chromosome 3p losses in head and neck cancers. AB - The interaction between environmental exposures and host susceptibility may lead to specific mutational events within head and neck squamous cell carcinoma (HNSCC). Furthermore, this interplay may determine not only the probability of cancer development but also the biologic characteristics of the tumor once it occurs. To better understand the relationship of mutagen sensitivity and tobacco and/or alcohol consumption on HNSCC carcinogenesis, we examined loss of heterozygosity on chromosome 3p in 58 HNSCCs using 10 microsatellite markers. Mutagen sensitivity was determined in vitro by quantitating bleomycin-induced chromatid breaks utilizing peripheral blood lympocytes from respective patients. Forty-six of the 58 invasive cancers showed allelic loss at one or more loci. Consistent with previous investigations, three discrete regions of deletions were identified: 3p13-14.2, 3p21.1-21. 2, and 3p25.1-26.1. The frequency and types of deletions were dependent upon tobacco and alcohol exposures. The distal region of 3p but not the remaining two regions was most frequently influenced by tobacco exposure. In contrast, heavy alcohol use when combined with tobacco use was associated with whole-arm loss of 3p rather than identifiable site-specific damage. Furthermore, this combined influence of alcohol and tobacco exposures on whole-arm loss was most apparent in those patients who expressed mutagen sensitivity; the odds ratio of whole-arm loss increasing from 2.67 (95% CI 0. 21 33.49) in those individuals who were mutagen resistant to 13.5 (95% CI 1.3-136.0; P = 0.02 by Fisher's exact test) in those who were mutagen sensitive. An assessment of clinical parameters in this population demonstrated that patients with whole-arm loss were more likely to present with cervical lymph node metastases and advanced stage disease than patients with partial losses. Results indicate that various environmental exposures as well as the expression of mutagen sensitivity will influence the types of chromosome 3p allelic losses in head and neck cancers as well as the behavior of disease once it develops. PMID- 10837017 TI - Isoflavonoids and lignans have different potentials to modulate oxidative genetic damage in human colon cells. AB - Polyphenolic compounds, including isoflavonoids and lignans, have been suggested to be chemopreventive on account of antioxidative properties. In this context it is of importance to have knowledge of their ability to reduce oxidative stress within target cells of tumorigenesis. Therefore, we investigated isoflavonoids and lignans for modulation of oxidative genetic damage in mammalian cells. H(2)O(2)-induced damage as well as endogenous DNA strand breaks and oxidized bases were determined after 30 min incubation of human colon cells with polyphenols using various modifications of the microgel electrophoresis assay (Comet assay). Enterolactone, a mammalian metabolite of plant lignans, was additionally investigated for modulation of intracellular oxidative stress in NIH 3T3 cells using laser scanning microscopy. In vivo effects of rye crispbread (a source of lignans) were investigated in 12 human volunteers by determining genetic damage in lymphocytes and antioxidant activity in plasma (FRAP assay). Genistein induced DNA breaks in the human tumour cell line HT29 clone 19A (12.5 100 microM). The polyphenols (100 microM) did not reduce damage induced by 150 microM H(2)O(2), indicating that they lacked antioxidative potential. At this concentration enterolactone also had no effect on intracellular oxidative stress induced by 31.25 and 125 microM H(2)O(2). In contrast, enterolactone, dihydrogenistein and formononetin reduced endogenous oxidative DNA damage at 100 microM. Daily ingestion of nine slices (76.5 g/day) of rye crispbread per day (containing 41.8 and 33.0 microg/100 g dry weight secoisolariciresinol and matairesinol, respectively) for 2 weeks did not significantly reduce genetic damage in blood lymphocytes, nor was there a modulation of plasma antioxidant capacity. The moderate effects of high concentrations of the tested compounds on endogenous oxidative DNA damage and failure to prevent H(2)O(2)- induced damage are indicative of only marginal protective potential by antioxidant mechanisms. The genotoxic effects of genistein deserve further investigation. PMID- 10837018 TI - 8,9-dihydroxy-8,9-dihydrodibenzo[a,l]pyrene is a potent morphological cell transforming agent in C3H10T(1)/(2)Cl8 mouse embryo fibroblasts in the absence of detectable stable covalent DNA adducts. AB - The comparative genotoxic effects of racemic trans-8,9-dihydroxy-8, 9 dihydrodibenzo[a,l]pyrene (trans-DB[a,l]P-8,9-diol), the metabolic K-region dihydrodiol of dibenzo[a,l] pyrene (DB[a,l]P) (dibenzo[def, p]chrysene) and DB[a,l]P in transformable mouse embryo C3H10T(1)/(2)Cl8 (C3H10T(1)/(2)) fibroblasts was investigated. The C3H10T(1)/(2) mouse embryo morphological cell transforming activities of these polycyclic aromatic hydrocarbons (PAHs) were assayed using concentration-response studies. At concentrations of 33 nM and above both trans-DB[a,l]P-8,9-diol and DB[a,l]P produced significant (and similar) numbers of type II and III foci per dish and numbers of dishes with type II and II foci. Concomitant cytotoxicity studies revealed a reduction in colony survival of approximately 25% up to 198 nM for both PAHs. DNA adducts of trans DB[a,l]P-8,9-diol and DB[a,l]P in C3H10T(1)/(2) cells were analyzed by a (32)P post-labeling TLC/HPLC method. No adducts were observed in the DNA of C3H10T(1)/(2) cells treated with trans-DB[a, l]P-8,9-diol at concentrations that induced morphological cell transformation. Under the same exposure and chromatographic conditions, DNA adducts of deoxyadenosine and deoxyguanosine derived from the fjord region anti-DB[a,l]P-11,12-diol-13,14-epoxide and syn DB[a,l]P-11,12-diol-13,14-epoxide were observed in the DNA of DB[a,l]P-treated cells. These results indicate that trans-DB[a,l]P-8, 9-diol has intrinsic genotoxic activity equal to that of DB[a,l]P, based on morphological cell transformation of mouse embryo fibroblasts. The activity of trans-DB[a,l]P-8,9 diol is apparently not associated with the formation of observable stable covalent DNA adducts. These results suggest that under appropriate conditions, trans-DB[a,l]P-8,9-diol may serve as an intermediate in the genotoxicity of DB[a,l]P. PMID- 10837019 TI - Tumors arising in DNA mismatch repair-deficient mice show a wide variation in mutation frequency as assessed by a transgenic reporter gene. AB - We reported previously that thymic lymphomas arising in mice lacking the DNA mismatch repair (MMR) gene, Msh2(-/-), exhibited striking elevations in the mutation frequency of a transgenic lacI reporter gene when compared with normal Msh2(-/-) tissues. To investigate whether hypermutation was a feature of all tumors arising in MMR-deficient mice, lacI transgene mutation frequencies were obtained from several different mouse tumors deficient for PMS2 and/or MSH2. While lacI gene hypermutation was again clearly evident in Msh2 +/- ms2(-/-) and Msh2(-/-)Pms2(-/-) thymic lymphomas, three non-thymic MSH2-deficient tumors failed to show lacI gene mutation frequency elevations when compared with a normal tissue of MMR-deficient mice. The elevated mutation frequencies in the lymphoid tumors, and the finding of multiple clustered mutations in lacI genes rescued from these tumors, suggest that they are possibly generated by a lymphoma specific hypermutational mechanism. PMID- 10837020 TI - Mice deficient in the nucleotide excision repair gene XPA have elevated sensitivity to benzo[a]pyrene induction of lung tumors. AB - This study is focused on chemical induction of lung tumors in xeroderma pigmentosum group A gene (XPA)-deficient mice to clarify the role of nucleotide excision repair (NER) in internal organs. Six-week-old female XPA-/-, XPA(+/-) and XPA(+/+) mice were instilled intratracheally with benzo[a] pyrene (B[a]P). A total of 68 surviving XPA mice treated with B[a]P were examined at month 16. The pulmonary adenoma incidence in XPA(-/-) mice was significantly higher than that in XPA(+/+) mice (71 versus 35%). Similarly, tumor multiplicity was elevated and, in addition, only XPA(-/-) mice had lung carcinomas. These results provide the first evidence that a deficiency in the NER gene XPA leads to enhanced tumorigenesis in the lung after exposure to B[a]P. PMID- 10837021 TI - Biotransformation of the tobacco-specific carcinogen 4-(methylnitrosamino)-1-(3 pyridyl)-1-butanone (NNK) in freshly isolated human lung cells PMID- 10837022 TI - Transcriptional regulation of adipogenesis. PMID- 10837023 TI - The protein encoded by the proto-oncogene DEK changes the topology of chromatin and reduces the efficiency of DNA replication in a chromatin-specific manner. AB - The structure of chromatin regulates the genetic activity of the underlying DNA sequence. We report here that the protein encoded by the proto-oncogene DEK, which is involved in acute myelogenous leukemia, induces alterations of the superhelical density of DNA in chromatin. The change in topology is observed with chromatin but not with naked DNA and does not involve dissociation of core histones from chromatin. Moreover, these effects require histone H2A/H2B dimers in addition to histone H3/H4. We additionally tested whether the DEK protein affects DNA-utilizing processes and found that the DEK protein substantially reduces the replication efficiency of chromatin but not of naked DNA templates. PMID- 10837024 TI - Dll4, a novel Notch ligand expressed in arterial endothelium. AB - We report the cloning and characterization of a new member of the Delta family of Notch ligands, which we have named Dll4. Like other Delta genes, Dll4 is predicted to encode a membrane-bound ligand, characterized by an extracellular region containing several EGF-like domains and a DSL domain required for receptor binding. In situ analysis reveals a highly selective expression pattern of Dll4 within the vascular endothelium. The activity and expression of Dll4 and the known actions of other members of this family suggest a role for Dll4 in the control of endothelial cell biology. PMID- 10837025 TI - gamma-catenin is regulated by the APC tumor suppressor and its oncogenic activity is distinct from that of beta-catenin. AB - beta-Catenin and gamma-catenin (plakoglobin), vertebrate homologs of Drosophila armadillo, function in cell adhesion and the Wnt signaling pathway. In colon and other cancers, mutations in the APC tumor suppressor protein or beta-catenin's amino terminus stabilize beta-catenin, enhancing its ability to activate transcription of Tcf/Lef target genes. Though beta- and gamma-catenin have analogous structures and functions and like binding to APC, evidence that gamma catenin has an important role in cancer has been lacking. We report here that APC regulates both beta- and gamma-catenin and gamma-catenin functions as an oncogene. In contrast to beta-catenin, for which only amino-terminal mutated forms transform RK3E epithelial cells, wild-type and several amino-terminal mutated forms of gamma-catenin had similar transforming activity. gamma-Catenin's transforming activity, like beta-catenin's, was dependent on Tcf/Lef function. However, in contrast to beta-catenin, gamma-catenin strongly activated c-Myc expression and c-Myc function was crucial for gamma-catenin transformation. Our findings suggest APC mutations alter regulation of both beta- and gamma-catenin, perhaps explaining why the frequency of APC mutations in colon cancer far exceeds that of beta-catenin mutations. Elevated c-Myc expression in cancers with APC defects may be due to altered regulation of both beta- and gamma-catenin. Furthermore, the data imply beta- and gamma-catenin may have distinct roles in Wnt signaling and cancer via differential effects on downstream target genes. PMID- 10837026 TI - E-MAP-115, encoding a microtubule-associated protein, is a retinoic acid inducible gene required for spermatogenesis. AB - Cell type-specific microtubules, such as the Sertoli cell microtubules and the manchette and flagellum microtubules of the spermatids, play essential roles in spermatogenesis. We identified the gene encoding E-MAP-115 (epithelial microtubule-associated protein of 115 kD) as a retinoic acid-inducible gene using gene trap mutagenesis in mouse embryonic stem cells. The gene trap insertion led to a null allele of the E-MAP-115 gene and, in agreement with its high expression in the testis, male mice homozygous for the mutation were sterile because of deformation of spermatid nuclei and subsequent gradual loss of germ cells. Consistent with a possible role for E-MAP-115 in stabilizing microtubules, microtubule associations in the mutant were morphologically abnormal in the manchette of spermatids and in Sertoli cells. We hypothesize that the abnormal microtubules in these two cell types are responsible for deformation of spermatid nuclei and germ cell loss, respectively, and indicate an essential role for E-MAP 115 in microtubule functions required for spermatogenesis. PMID- 10837027 TI - Notch signaling is essential for vascular morphogenesis in mice. AB - The Notch gene family encodes large transmembrane receptors that are components of an evolutionarily conserved intercellular signaling mechanism. To assess the role of the Notch4 gene, we generated Notch4-deficient mice by gene targeting. Embryos homozygous for this mutation developed normally, and homozygous mutant adults were viable and fertile. However, the Notch4 mutation displayed genetic interactions with a targeted mutation of the related Notch1 gene. Embryos homozygous for mutations of both the Notch4 and Notch1 genes often displayed a more severe phenotype than Notch1 homozygous mutant embryos. Both Notch1 mutant and Notch1/Notch4 double mutant embryos displayed severe defects in angiogenic vascular remodeling. Analysis of the expression patterns of genes encoding ligands for Notch family receptors indicated that only the Dll4 gene is expressed in a pattern consistent with that expected for a gene encoding a ligand for the Notch1 and Notch4 receptors in the early embryonic vasculature. These results reveal an essential role for the Notch signaling pathway in regulating embryonic vascular morphogenesis and remodeling, and indicate that whereas the Notch4 gene is not essential during embryonic development, the Notch4 and Notch1 genes have partially overlapping roles during embryogenesis in mice. PMID- 10837028 TI - Dimerization and nuclear entry of mPER proteins in mammalian cells. AB - Nuclear entry of circadian oscillatory gene products is a key step for the generation of a 24-hr cycle of the biological clock. We have examined nuclear import of clock proteins of the mammalian period gene family and the effect of serum shock, which induces a synchronous clock in cultured cells. Previously, mCRY1 and mCRY2 have been found to complex with PER proteins leading to nuclear import. Here we report that nuclear translocation of mPER1 and mPER2 (1) involves physical interactions with mPER3, (2) is accelerated by serum treatment, and (3) still occurs in mCry1/mCry2 double-deficient cells lacking a functional biological clock. Moreover, nuclear localization of endogenous mPER1 was observed in cultured mCry1/mCry2 double-deficient cells as well as in the liver and the suprachiasmatic nuclei (SCN) of mCry1/mCry2 double-mutant mice. This indicates that nuclear translocation of at least mPER1 also can occur under physiological conditions (i.e., in the intact mouse) in the absence of any CRY protein. The mPER3 amino acid sequence predicts the presence of a cytoplasmic localization domain (CLD) and a nuclear localization signal (NLS). Deletion analysis suggests that the interplay of the CLD and NLS proposed to regulate nuclear entry of PER in Drosophila is conserved in mammals, but with the novel twist that mPER3 can act as the dimerizing partner. PMID- 10837029 TI - Tissue- and stage-specific modulation of Wingless signaling by the segment polarity gene lines. AB - Wnt signaling controls a variety of developmental programs but the mechanisms by which the same signal leads to distinct outputs remain unclear. To address this question, we identified stage-specific modulators of Wingless (Wg) signaling in the Drosophila embryonic epidermis. We show that lines (lin) is essential for Wg dependent patterning in dorsal epidermis. lin encodes a novel protein that acts cell-autonomously, downstream or in parallel to Armadillo (Arm) and upstream of Wg-dependent target genes. Lin can accumulate in nuclei of cells signaled by Wg, suggesting that signaling promotes entry of Lin into the nucleus, where it cooperates with Arm and Pangolin. Thus, a stage-specific modulator is used to mediate Wg signaling activity in dorsal patterning. Hedgehog (Hh) controls half of the parasegmental pattern dorsally and antagonizes Wg function to do so. Lin can accumulate in the cytoplasm of cells signaled by Hh, suggesting that Hh antagonizes Wg function by prohibiting Lin from entering the nucleus. PMID- 10837030 TI - Two lineage boundaries coordinate vertebrate apical ectodermal ridge formation. AB - Proximal-distal outgrowth of the vertebrate limb bud is regulated by the apical ectodermal ridge (AER), which forms at an invariant position along the dorsal ventral (D/V) axis of the embryo. We have studied the genetic and cellular events that regulate AER formation in the mouse. In contrast to implications from previous studies in chick, we identified two distinct lineage boundaries in mouse ectoderm prior to limb bud outgrowth using a Cre/loxP-based fate-mapping approach and a novel retroviral cell-labeling technique. One border is transient and at the limit of expression of the ventral gene En1, which corresponds to the D/V midline of the AER, and the second border corresponds to the dorsal AER margin. Labeling of AER precursors using an inducible Cre showed that not all cells that initially express AER genes form the AER, indicating that signaling is required to maintain an AER phenotype. Misexpression of En1 at moderate levels specifically in the dorsal AER of transgenic mice was found to produce dorsally shifted AER fragments, whereas high levels of En1 abolished AER formation. In both cases, the dorsal gene Wnt7a was repressed in cells adjacent to the En1 expressing cells, demonstrating that signaling regulated by EN1 occurs across the D/V border. Finally, fate mapping of AER domains in these mutants showed that En1 plays a part in positioning and maintaining the two lineage borders. PMID- 10837031 TI - N-myc can functionally replace c-myc in murine development, cellular growth, and differentiation. AB - Members of the myc family of cellular oncogenes have been implicated as transcriptional regulators in pathways that govern cellular proliferation and death. In addition, N-myc and c-myc are essential for completion of murine embryonic development. However, the basis for the evolutionary conservation of myc gene family has remained unclear. To elucidate this issue, we have generated mice in which the endogenous c-myc coding sequences have been replaced with N-myc coding sequences. Strikingly, mice homozygous for this replacement mutation can survive into adulthood and reproduce. Moreover, when expressed from the c-myc locus, N-myc is similarly regulated and functionally complementary to c-myc in the context of various cellular growth and differentiation processes. Therefore, the myc gene family must have evolved, to a large extent, to facilitate differential patterns of expression. PMID- 10837032 TI - Gross chromosomal rearrangements and genetic exchange between nonhomologous chromosomes following BRCA2 inactivation. AB - Cancer-causing mutations often arise from gross chromosomal rearrangements (GCRs) such as translocations, which involve genetic exchange between nonhomologous chromosomes. Here we show that murine Brca2 has an essential function in suppressing GCR formation after chromosome breakage. Cells that harbor truncated Brca2 spontaneously incur GCRs and genomic DNA breaks during division. They exhibit hypersensitivity to DNA damage by interstrand cross-linkers, which even at low doses trigger aberrant genetic exchange between nonhomologous chromosomes. Therefore, genetic instability in Brca2-deficient cells results from the mutagenic processing of spontaneous or induced DNA damage into gross chromosomal rearrangements, providing a mechanistic basis for cancer predisposition. PMID- 10837034 TI - Vaccine procurement and self-sufficiency in developing countries. AB - This paper discusses the movement toward self-sufficiency in vaccine supply in developing countries (and countries in transition to new economic and political systems) and explains special supply concerns about vaccine as a product class. It traces some history of donor support and programmes aimed at self-financing, then continues with a discussion about self-sufficiency in terms of institutional capacity building. A number of deficiencies commonly found in vaccine procurement and supply in low- and middle-income countries are characterized, and institutional strengthening with procurement technical assistance is described. The paper also provides information about a vaccine procurement manual being developed by the United States Agency for International Development (USAID) and the World Health Organization (WHO) for use in this environment. Two brief case studies are included to illustrate the spectrum of existing capabilities and different approaches to technical assistance aimed at developing or improving vaccine procurement capability. In conclusion, the paper discusses the special nature of vaccine and issues surrounding potential integration and decentralization of vaccine supply systems as part of health sector reform. PMID- 10837033 TI - Dictyostelium RasD is required for normal phototaxis, but not differentiation. AB - RasD, a Dictyostelium homolog of mammalian Ras, is maximally expressed during the multicellular stage of development. Normal Dictyostelium aggregates are phototactic and thermotactic, moving towards sources of light and heat with great sensitivity. We show that disruption of the gene for rasD causes a near-total loss of phototaxis and thermotaxis in mutant aggregates, without obvious effects on undirected movement. Previous experiments had suggested important roles for RasD in development and cell-type determination. Surprisingly, rasD(-) cells show no obvious changes in these processes. These cells represent a novel class of phototaxis mutant, and indicate a role for a Ras pathway in the connections between stimuli and coordinated cell movement. PMID- 10837035 TI - Priority setting for health research: lessons from developing countries. The Working Group on Priority Setting. AB - Research resources for addressing health problems of developing countries remain disproportionately low compared with the tremendous disease burdens borne by these countries. There is a need to focus these scarce resources on research that will optimize health benefits and lead to equity. This paper reviews processes and methods that have been used for setting research priorities. Past and current processes have focused on expert-driven research agenda, emphasizing scientific autonomy and global analyses. Methods for setting priorities have focused on the metrics of disease burdens, while less attention has been placed on who sets priorities and how choices are made. The paper proposes a strategy of priority setting, based on lessons learned from essential national health research (ENHR) approaches attempted in several developing countries. With equity in health and development as its goal, the proposed model is demand-driven, and involves multi dimensional inputs and multiple stakeholders. Various steps of the process are discussed: getting participants involved; gathering evidence and information; determining criteria for priority setting; and implementation and evaluation. The paper concludes with a discussion of the gap between national research priorities and the research agenda set at regional and global levels, an issue that needs to be satisfactorily addressed in the future. PMID- 10837036 TI - Young people's health in developing countries: a neglected problem and opportunity. AB - Although the number of young people in developing countries is increasing, their health has not been considered a priority. This is partly because of their lower mortality relative to other age groups, but also because of a lack of appropriate data collection and analysis. However, the burden of disease in young people is not trivial, and most importantly, it is during adolescence that long-term health related behaviours are formed. Most research investigating health problems in young people has tended to concentrate on a narrow conception of problem areas, such as those relating to reproduction and substance abuse. However, in many developing countries diseases such as schistosomiasis and intestinal parasites may also have a disproportionate impact on young people's health. Young people often have radically different perceptions and priorities in terms of health and disease to those of health planners. Successful programmes will need to incorporate the views of young people themselves. Interventions targeted at young people have enormous potential for primary and secondary prevention of a huge range of health problems and present an opportunity that should not be ignored. PMID- 10837037 TI - The Nepal National Vitamin A Program: prototype to emulate or donor enclave? AB - More than 250 million of the world's children suffer from vitamin A deficiency. Nepal is one of 60 countries in which this deficiency constitutes a significant public health problem. Each year in Nepal, vitamin A deficiency is responsible for the deaths of 9000 children and for 2500 children becoming permanently blind. The Nepal National Vitamin A Program (NVAP) was begun in 1993 in eight of the country's 75 districts. By the end of 1997, the programme covered 32 districts, and by 2003 its coverage will be nationwide. The Nepal NVAP is considered by many to be a highly successful, model programme. It consists primarily of distributing high-dose vitamin A capsules to all children 6 to 60 months of age during twice yearly campaigns. The capsule distribution is carried out by a previously existing network of Female Community Health Volunteers (FCHVs) that has been reinvigorated by the highly visible and universally acclaimed success of the NVAP. An important strategy of the programme has been the empowerment of the FCHVs, which has been accomplished by organizing, training and motivating community workers and other representatives from education, agriculture and other sectors, as well as political representatives, to support the FCHVs. The annual cost of the NVAP is US$1.7 million. It costs $1.25 to deliver two vitamin A capsules to each participant. The cost per averted death is $327. The NVAP reduces the incidence and severity of diarrhoeal disease and measles, which in turn reduces the need for Ministry of Health services, thereby annually saving the Government of Nepal $1.5 million. Factoring in these cost savings, the net annual cost of the current NVAP is $167,000, and the net annual cost of the permanent, nationwide programme is estimated at $1.1 million. The NVAP is a highly cost-effective programme. The article concludes with a discussion of the sustainability and replicability of the programme. PMID- 10837038 TI - The Chinese experience of hospital price regulation. AB - This paper analyzes the distortion effects of the hospital pricing policies in China. To help maintain equitable access to hospital services, the Chinese government regulates prices of hospital services, and provides subsidies to public hospitals. Comparing the regulated fees of selected hospital services with their average unit costs indicates that the average cost-recovery rate of the fees is only 50%. The fees for 90% of the services are less than their average unit costs, while the fees for the high-tech services exceed their costs. Moreover, the State Price Commission allowed a drug profit margin of 15-20% over the wholesale price. The distorted fee schedule affects the behaviour of hospitals. Empirical evidence revealed problems of violation of price regulations (charging a fee exceeding the regulated fee), over-provision of profitable high tech services and over-prescription of drugs. The Chinese experience shows that low regulated fees cannot reduce the economic burden on patients, and that distorted medical fees can result in distorted service provision and low efficiency of medical resources. Strategies to correct for the price distortions are discussed. PMID- 10837039 TI - Public health in Russia: the view from the inside. AB - The health of the Russian population continues to lag far behind that in the west. A robust public health response to the high levels of communicable and non communicable diseases is required. This challenge has attracted considerable attention from international donor agencies and others, but there are still many questions about how the health situation in Russia is understood by policy-makers within the country and what responses are being considered. This paper examines these questions by means of a review of literature published in Russia and interviews with key informants. It concludes that although many of the determinants of health in Russia have been identified, they are typically discussed in a general way. Research on the major determinants of disease in Russia, and published in the international literature, appears to have had little impact. The need for reform to enhance the public health response is recognized. Goals of reform have been described but are poorly defined and there is typically little relationship between a stated goal and the strategy proposed to achieve it. There is a lack of clarity about what is meant by public health, and key concepts, such as inter-sectoral and multi-disciplinary working, are either ignored or misunderstood. Evidence of capacity for managed change is weak. There is an urgent need to create a shared awareness of evidence on the nature of the health challenges facing Russia and the evidence base for both the content of potential responses and the strategies that might be adopted to implement them. PMID- 10837041 TI - Rhetoric or reality? The participation of disabled people in NGO planning. AB - User involvement in planning is now well established in the fields of both health and development. This study looks at one particular client group, namely disabled people, and addresses the question: How do selected European-based international non-governmental organizations (INGOs) facilitate the participation of disabled people in their planning process? The study was exploratory in nature, using both quantitative and qualitative methods to answer the research question. Data were collected using a self-administered questionnaire with 18 INGOs, which was supported by five semi-structured interviews and 20 'e-mail dialogues' with key informants. The findings indicate that the INGOs involve disabled people's organizations (DPOs) in their planning of services and projects in a variety of ways. Most commonly this is through sharing information with them, but consulting them, including them in decision-making or supporting action initiated by them are other less frequent methods of involvement. INGOs with a specific disability focus involve disabled people in the ways described above more frequently than other INGOs. Although most INGOs regularly provide information and consult DPOs, if there is no assurance that ideas raised will be implemented, then there is no guarantee of DPOs' participation in the planning process of these INGOs. The focus of an INGO and the nature of its projects affect how disabled people are involved in planning, whereas the size of an INGO has little effect. INGOs thus far have failed to match their expressed intentions about participation, but as they help to strengthen DPOs, encourage their formation and move to make disability an issue that cuts across sectoral boundaries, INGOs are changing. There is still a lot of rhetoric about participation, but the participation of disabled people in the planning process of INGOs is a growing reality. PMID- 10837040 TI - Information quality in a remote rural maternity unit in Ghana. AB - The collection of accurate maternal outcome data enables causes of morbidity and mortality to be identified, which in turn permits interventions to be targeted appropriately. It also allows estimates to be made about the importance of various indicators in predicting birth outcome. These indicators can then be compared between health services, across time and against programme objectives, thus ensuring a management information system that informs policy and provides for real change. A review was done of data collection at the antenatal clinic and maternity ward in a remote rural hospital in northern Ghana. The data collected came from maternity ward records and participant observation, and they highlight deficiencies in the record management procedures. It is argued that exhorting staff to greater accuracy, although obvious, may not be the only solution, because of the structural impediments that often give an illusion of accuracy. The best data need to be collected within the constraints of the equipment and the people. Furthermore, to make the task more meaningful, regular feedback needs to be provided so that the process of record keeping is relevant to those who do it. Ministries of health need to conduct regular audits, like this microanalysis, so that policies are not based on data that are analyzed under a presumption of accuracy. PMID- 10837042 TI - Attitudes to 'Kaponya Mafumo': the terminators of pregnancy in urban Zambia. AB - As part of a larger study of adolescent sexual and reproductive health in urban Zambia, the issue of unwanted pregnancy and abortion was considered through the examination of the perceptions of both adolescents and adults. Young people rank sexual health as their primary health issue, and sexual behaviour is integrally linked into other aspects of their lives. Pregnancies were deemed to be a common occurrence amongst the adolescents, with an estimated two-thirds of unwanted pregnancies ending in unsafe abortion. The decision to abort is primarily determined by the reaction of the boyfriend and his willingness to accept paternity and the associated financial implications. Other crucial influences are the desire to stay in school and the stigma attached to unwanted pregnancy. The decision-making process regarding the abortion itself is related to the perceived advantages and disadvantages of various service providers. Around 40% of the respondents stated that in the event of an abortion being carried out, it would be performed either by the girl herself or with the assistance of other non medical personnel. Less popular but still significant are traditional healers and private doctors. Formal health services tend to be rejected due to their poor perception by young people, centred on the lack of privacy and confidentiality, and the de facto illegal nature of abortion itself. The services of nurses are sought, but outside of the clinic setting. The most popular method of self induced abortion is overdosing on chloroquine. Other methods involve the use of traditional medicines such as various types of roots, as well as more modern methods such as ingesting washing powder. Recommendations for policy-makers concentrate on the improvement of formal, 'youth friendly' health services and the development of appropriate outreach education methods which address specific concerns widely held by young people. PMID- 10837043 TI - Conceptualizing and applying a minimum basic needs approach in southern Philippines. AB - This study, a collaboration between Canadian and Filipino researchers, focuses on how the national government's Minimum Basic Needs (MBN) Approach has been implemented at the local level in some selected sites in Region XI on the Philippine island of Mindanao. This case study of MBN implementation focuses on the experiences of three municipalities and three barangays (villages) within them. The research explores, through interviews and group discussions, what the mayors, technical working groups and volunteer health workers in these areas thought about MBN and how they participated in the initiative. The objectives of the study were: to explore models of MBN data utilization at the municipal and barangay levels; to understand how the MBN data guided decision-making about community priorities and resource allocation; to examine the role that community volunteers played in promoting the use of MBN data, and in community health and development activities which ensued; and to determine what factors challenged or encouraged the use of MBN data for social development at the barangay level. In all the sites, MBN had some impact, most often due to methods of concentrating information on unmet basic needs locally and making use of it in planning and project development processes. The findings show that although there is still some way to go before MBN is effectively integrated into local planning and project development, some responses to problems have been implemented and innovative projects were undertaken or being considered. PMID- 10837044 TI - Universal health care? The views of Negev Bedouin Arabs on health services. AB - BACKGROUND: This study examines health and health care attitudes, practices and utilization patterns among the Bedouin Arab minority in the south of Israel. Particular attention is given to the effects of the new National Insurance Law that provides universal coverage for the first time, and to the identification of critical issues for further research. METHODS: Focus groups, adapted to Bedouin culture, were the primary method of data collection. Twelve groups (158 participants) from throughout the Negev met for 3-7 sessions each, using specially trained local moderators and observers. Issues discussed and analyzed included experience and satisfaction with the current health system (both modern and traditional), health service availability/barriers, health care needs, influences of social change, and the National Insurance Law. RESULTS: Participants voice dissatisfaction with modern health services in the Bedouin sector and the state of health of Negev Bedouin. They place great emphasis on the connection between health and the rapid social and economic changes, which this traditionally nomadic group is undergoing. Traditional health care is felt to still exist, but its importance is waning. The National Insurance law is having a major impact on the Bedouin, particularly because it provides universal health insurance coverage where only partial coverage had been in effect. CONCLUSIONS: This study, one of the first of its kind in the Bedouin sector, showed that the focus group method, if properly modified to cultural norms, can be a valuable research tool in traditional communities and in health service research. The findings from this research can be used to direct efforts to improve health policy and health services for this group, as well as preparing the way for further qualitative or quantitative studies. PMID- 10837046 TI - Maternal and child health services in rural Nepal: does access or quality matter more? AB - This study seeks to establish the relative importance of service access and quality on utilization of preventive health services in the western and middle western Hill region of Nepal. Access was measured in terms of travel time to the nearest health post and coverage by outreach workers. The quality of static services was defined in structural terms: physical infrastructure, number of staff, availability of drugs and holding of special maternal and child health clinics. The initial analysis showed that no single indicator of quality was of overriding importance and therefore an overall quality index was constructed. After adjustment for access and for socioeconomic characteristics of families and communities, a very pronounced relationship between overall structural quality of the nearest health post and service uptake persisted. The adjusted odds of using some form of antenatal service were 6.6 times higher in the catchment areas of high quality posts than in areas served by low quality posts. The corresponding figure for receipt of BCG vaccination is 8.1. By comparison, the effects of travel time to the nearest health post are modest. Uptake of services is about twice as high when there is a health post in the community. Regular monthly visits by outreach workers also had a marked effect on service utilization. These results suggest that investment in the quality of health posts is more important than further increases in their number and that a further expansion of outreach services is a priority. PMID- 10837045 TI - Evaluation of blood bank practices in Karachi, Pakistan, and the government's response. AB - BACKGROUND: National legislation in Pakistan regulating blood banks has been introduced several times, but has never been passed. To support provincial-level efforts to develop legislation we conducted a study to evaluate blood-banking practices in Karachi, Pakistan, to identify areas that could be improved. METHODS: Thirty-seven blood banks were randomly selected from a list of 87 Karachi blood banks. The research team interviewed blood bank personnel, inspected available facilities and equipment, and observed blood collection using structured questionnaires and observation forms. RESULTS: Of the 37 selected facilities, 25 were operational and 24 agreed to participate. Twelve (50%) of the facilities reported regularly utilizing paid blood donors, while only six (25%) activity recruited volunteer donors. During observation only 8% of facilities asked donors about injecting drug use, and none asked donors any questions about high-risk sexual behaviour. While 95% of blood banks had appropriate equipment and reagents to screen for hepatitis B, only 55% could screen for HIV and 23% for hepatitis C. Twenty-nine percent of the facilities were storing blood products outside the WHO recommended temperature limits. IMPLICATIONS: Practices at most Karachi blood banks fell well below WHO standards. Findings from this study were instrumental in developing and passing legislation to regulate blood transfusion throughout Sindh Province, and suggest a method for improving blood transfusion practices in other developing countries. PMID- 10837047 TI - The real and the nominal? Making inflationary adjustments to cost and other economic data. AB - Given the scarcity of cost data for health interventions, there has been substantial use of a relatively small number of existing studies to underpin policy development formulation. Intervention-specific cost and cost-effectiveness data have been used to plan overall budgets, to assess the relative efficiency of different interventions and to consider the resource requirements for programme implementation at both the local and national levels. Cost and cost-effectiveness comparisons have been made between these studies and general sources such as the World Bank's World Development Report 1993. At the same time, information on key health sector variables, such as annual health expenditures, has been systematically compiled for more than two decades. The question of possible inflationary effects is becoming increasingly important as the original data on which these numbers are based ages. For example, cost figures from the mid-1980s require a 60% inflationary adjustment simply to maintain their real value in current dollars. This paper looks at methods to adjust cost data to account for inflation and discusses the difference between real or constant and nominal or current values. These methods are also used to make inflationary adjustments to other types of economic data such as income. PMID- 10837048 TI - International health aid and professional independence: the need for quality standards and self-regulation. PMID- 10837049 TI - Conferences. AB - International Conference on Health Research for Development * 10-13 October 2000 * Bangkok, Thailand 128th Annual Meeting of the American Public Health Association: Eliminating Health Disparities * 12-16 November 2000 * Boston, MA, USA PMID- 10837050 TI - Discovering the role of the major histocompatibility complex in the immune response. AB - The discovery that genes in the major histocompatibility complex (MHC) play an important role in the immune response depended on the chance interaction of several unrelated events. The first, and most important, was the decision by Michael Sela to synthesize a series of branched, multichain, synthetic polypeptides based on a backbone of poly-l-lysine. The prototype compound, (T,G) A-L, was tipped with short random sequences of tyrosine and glutamic acid. This resulted in a restricted range of antigenic determinants composed of only two or three amino acids with a variable length-ideal for binding to the peptide binding groove of MHC class II molecules. The second was the decision by John Humphrey to immunize various strains of rabbits with this synthetic polypeptide. Two of these rabbit strains showed very large quantitative differences in antibody response to (T, G)-A-L. In transferring this system to inbred mouse strains, the third bit of good fortune was the availability at the National Institute of Medical Research, in Mill Hill (London), of the CBA (H2(k)) and C57 (H2(b)) strains. The H2(b) haplotype is the only one mediating a uniform high antibody response to (T,G)-A L. The fourth critical ingredient was the availability of numerous congenic and H2 recombinant inbred strains of mice produced earlier by Snell, Stimpfling, Shreffler, and Klein. A search for congenic pairs of mice expressing the responder and nonresponder H2 haplotypes on the same background revealed that these strains responded as a function of their H2 haplotype, not of their inbred background. Extensive studies in a variety of inbred strains carrying recombinant H2 haplotypes, as well as a four-point linkage cross, mapped immune response to (T,G)A-L within the murine MHC, between the K and Ss loci. The demonstration that stimulation in the mixed lymphocyte reaction (MLR) mapped to the same region quickly led to attempts to produce antisera in congenic H2 recombinant strain combinations. These antisera identified I-region associated (Ia) antigens. Immunoprecipitation and blocking studies showed that the gene products controlling specific immune responses, the mixed lymphocyte reaction, and the structure of Ia antigens were one and the same-now designated as the I-A MHC class II molecules. These antisera and inbred strains enabled Unanue to demonstrate the peptide binding function of class II MHC molecules. PMID- 10837052 TI - Molecular basis of celiac disease. AB - Celiac disease (CD) is an intestinal disorder with multifactorial etiology. HLA and non-HLA genes together with gluten and possibly additional environmental factors are involved in disease development. Evidence suggests that CD4(+) T cells are central in controlling an immune response to gluten that causes the immunopathology, but the actual mechanisms responsible for the tissue damage are as yet only partly characterized. CD provides a good model for HLA-associated diseases, and insight into the mechanism of this disease may well shed light on oral tolerance in humans. The primary HLA association in the majority of CD patients is with DQ2 and in the minority of patients with DQ8. Gluten-reactive T cells can be isolated from small intestinal biopsies of celiac patients but not of non-celiac controls. DQ2 or DQ8, but not other HLA molecules carried by patients, are the predominant restriction elements for these T cells. Lesion derived T cells predominantly recognize deamidated gluten peptides. A number of distinct T cell epitopes within gluten exist. DQ2 and DQ8 bind the epitopes so that the glutamic acid residues created by deamidation are accommodated in pockets that have a preference for negatively charged side chains. Evidence indicates that deamidation in vivo is mediated by the enzyme tissue transglutaminase (tTG). Notably, tTG can also cross-link glutamine residues of peptides to lysine residues in other proteins including tTG itself. This may result in the formation of complexes of gluten-tTG. These complexes may permit gluten-reactive T cells to provide help to tTG-specific B cells by a mechanism of intramolecular help, thereby explaining the occurrence of gluten-dependent tTG autoantibodies that is a characteristic feature of active CD. PMID- 10837053 TI - Population biology of lymphocytes: the flight for survival. AB - In this essay we suggest that the primary goal of the cells of the immune system is to ensure their own growth and survival. In adults, in steady-state conditions, the number and distribution of lymphocyte populations is under homeostatic control. New lymphocytes that are continuously produced in primary and secondary lymphoid organs must compete with resident cells for survival. We discuss recent findings supporting lymphocyte survival as a continuous active process and implicating cognate receptor engagement as fundamental survival signals for both T and B lymphocytes. The conflict of survival interests between different cell types gives rise to a pattern of interactions that mimics the behavior of complex ecological systems. In their flight for survival and in response to competition, lymphocytes use different survival signals within different ecological niches during cell differentiation. This is the case for T and B lymphocytes and also for naive and memory/activated T and B cells. We discuss how niche differentiation allows the co-existence of different cell types and guarantees both repertoire diversity and efficient immune responses. PMID- 10837054 TI - Nonclassical MHC class II molecules. AB - Major histocompatibility complex (MHC) class II molecules are cell surface proteins that present peptides to CD4(+) T cells. In addition to these wellcharacterized molecules, two other class II-like proteins are produced from the class II region of the MHC, HLA-DM (DM) and HLA-DO (DO) (called H2-M, or H2 DM and H2-O in the mouse). The function of DM is well established; it promotes peptide loading of class II molecules in the endosomal/lysosomal system by catalyzing the release of CLIP peptides (derived from the class II-associated invariant chain) in exchange for more stably binding peptides. While DM is present in all class II- expressing antigen presenting cells, DO is expressed mainly in B cells. In this cell type the majority of DM molecules are not present as free heterodimers but are instead associated with DO in tight heterotetrameric complexes. The association with DM is essential for the intracellular transport of DO, and the two molecules remain associated in the endosomal system. DO can clearly modify the peptide exchange activity of DM both in vitro and in vivo, but the physiological relevance of this interaction is still only partly understood. PMID- 10837051 TI - Receptor selection in B and T lymphocytes. AB - The process of clonal selection is a central feature of the immune system, but immune specificity is also regulated by receptor selection, in which the fate of a lymphocyte's antigen receptor is uncoupled from that of the cell itself. Whereas clonal selection controls cell death or survival in response to antigen receptor signaling, receptor selection regulates the process of V(D)J recombination, which can alter or fix antigen receptor specificity. Receptor selection is carried out in both T and B cells and can occur at different stages of lymphocyte differentiation, in which it plays a key role in allelic exclusion, positive selection, receptor editing, and the diversification of the antigen receptor repertoire. Thus, the immune system takes advantage of its control of V(D)J recombination to modify antigen receptors in such a way that self/non-self discrimination is enhanced. New information about receptor editing in T cells and B-1 B cells is also discussed. PMID- 10837055 TI - Negative regulation of cytokine signaling pathways. AB - The Janus family of protein tyrosine kinases (JAKs) and STAT transcription factors regulate cellular processes involved in cell growth, differentiation, and transformation through their association with cytokine receptors. The CIS family of proteins (also referred to as the SOCS or SSI family) has been implicated in the regulation of signal transduction by a variety of cytokines. Most of them appear to be induced after stimulation with several different cytokines, and at least three of them (CIS1, CIS3/SOCS3, and JAB/SOCS1) negatively regulate cytokine signal transduction by various means: CIS1 inhibits STAT5 activation by binding to cytokine receptors that recruit STAT5, whereas JAB/SOCS-1 and CIS3/SOCS-3 directly bind to the kinase domain of JAKs, thereby inhibiting tyrosine-kinase activity. Therefore, these CIS family members seem to function in a classical negative feedback loop of cytokine signaling. Biochemical characterization as well as gene disruption studies indicate that JAB/SOCS1/SSI-1 is an important negative regulator of interferon gamma signaling. The mechanisms by which these inhibitors of cytokine signal transduction exert their effects have been extensively studied and will provide useful information for regulating tyrosine-kinase activity. PMID- 10837056 TI - T cell activation and the cytoskeleton. AB - Ligation of the T cell antigen receptor (TCR) stimulates protein tyrosine kinases (PTKs), which regulate intracellular calcium and control the activity of protein kinase C (PKC) isozymes. PTKs activated by antigen receptors and costimulatory molecules also couple to phosphatidylinositol-3 kinase (PI3K) and control the activity of Ras- and Rho-family GTPases. T cell signal transduction is triggered physiologically by antigen in the context of antigen presenting cells (APC). The formation of stable and prolonged contacts between T cells and APCs is not necessary to initiate T cell signaling but is required for effective T cell proliferation and differentiation. The stabilization of the T cell/ APC conjugate is regulated by intracellular signals induced by antigen receptors and costimulators. These coordinate the regulation of the actin and microtubule cytoskeleton and organize a specialized signaling zone that allows sustained TCR signaling. PMID- 10837057 TI - The specific regulation of immune responses by CD8+ T cells restricted by the MHC class Ib molecule, Qa-1. AB - Over the last three decades considerable evidence has accumulated that CD8(+) T cells regulate peripheral immune responses, in part, by specifically controlling the outgrowth of antigen-triggered CD4(+) T cells. This regulatory function of CD8(+) T cells has been shown, in vivo, to control the emergence of autoreactive CD4(+) T cells as well as CD4(+) T cells reactive to conventional antigens, including alloantigens. In this review, we summarize the evidence that this immune suppression mediated by CD8(+) T cells is dependent, in part, on specific cognate interactions between MHC class I-restricted regulatory CD8(+) cells and antigen-activated CD4(+) T cells. Moreover, we review the evidence that regulatory CD8(+) T cells recognize antigen-activated CD4(+) T cells in a TCR specific manner restricted by the MHC class Ib molecule, Qa-1. The Qa-1 molecule may be uniquely qualified to serve this MHC restrictive function because, unlike conventional MHC molecules, it is preferentially and transiently expressed on activated and not resting CD4(+) T cells. This may assure that only recently antigen-activated CD4(+) T cells expressing Qa-1/TCR peptide complexes will induce regulatory CD8(+) T cells and subsequently become susceptible to regulation. Because Qa-1 also binds to self Qdm peptides that trigger NK (CD94/ NKG2) receptors on CD8(+) T cells, the machinery for homeostatic regulation of regulatory CD8(+) T cells can be envisioned. Finally, we propose a model by which these TCR specific, Qa-1-restricted regulatory CD8(+) T cells selectively downregulate antigen-activated T cells expressing TCRs of certain affinities. Ultimately these regulatory CD8(+) T cells control the peripheral TCR repertoire during the course of immune responses to both self and foreign antigens. PMID- 10837058 TI - The biology of chemokines and their receptors. AB - During the last five years, the development of bioinformatics and EST databases has been primarily responsible for the identification of many new chemokines and chemokine receptors. The chemokine field has also received considerable attention since chemokine receptors were found to act as co-receptors for HIV infection (1). In addition, chemokines, along with adhesion molecules, are crucial during inflammatory responses for a timely recruitment of specific leukocyte subpopulations to sites of tissue damage. However, chemokines and their receptors are also important in dendritic cell maturation (2), B (3), and T (4) cell development, Th1 and Th2 responses, infections, angiogenesis, and tumor growth as well as metastasis (5). Furthermore, an increase in the number of chemokine/receptor transgenic and knock-out mice has helped to define the functions of chemokines in vivo. In this review we discuss some of the chemokines' biological effects in vivo and in vitro, described in the last few years, and the implications of these findings when considering chemokine receptors as therapeutic targets. PMID- 10837059 TI - Dendritic cells in cancer immunotherapy. AB - The potential to harness the potency and specificity of the immune system underlies the growing interest in cancer immunotherapy. One such approach uses bone marrow-derived dendritic cells, phenotypically distinct and extremely potent antigen-presenting cells, to present tumor-associated antigens and thereby generate tumor-specific immunity. Support for this strategy comes from animal studies that have demonstrated that dendritic cells, when loaded ex vivo with tumor antigens and administered to tumor-bearing hosts, can elicit T cell mediated tumor destruction. These observations have led to clinical trials designed to investigate the immunologic and clinical effects of antigen-loaded dendritic cells administered as a therapeutic vaccine to patients with cancer. In the design and conduct of such trials, important considerations include antigen selection, methods for introducing the antigen into MHC class I and II processing pathways, methods for isolating and activating dendritic cells, and route of administration. Although current dendritic cell-based vaccination methods are cumbersome, promising results from clinical trials in patients with malignant lymphoma, melanoma, and prostate cancer suggest that immunotherapeutic strategies that take advantage of the antigen presenting properties of dendritic cells may ultimately prove both efficacious and widely applicable to human tumors. PMID- 10837060 TI - CD8+ T cell effector mechanisms in resistance to infection. AB - Based on T cell subset depletion studies and the analysis of gene knockout mice, it is evident that CD8(+) T cells contribute to resistance against intracellular infections with certain viral, protozoan, and bacterial pathogens. Although they are known primarily for their capacity to kill infected cells, CD8(+) T cells elaborate a variety of effector mechanisms with the potential to defend against infection. Microbes use multiple strategies to cause infection, and the nature of the pathogenhost interaction may determine which CD8(+) T cell effector mechanisms are required for immunity. In this review, we summarize our current understanding of the effector functions used by CD8(+) T cells in resistance to pathogens. Analyses of mice deficient in perforin and/or Fas demonstrate that cytolysis is critical for immunity against some, but not all, infections and also reveal the contribution of cytolysis to the pathogenesis of disease. The role of CD8(+) T cell-derived cytokines in resistance to infection has been analyzed by systemic treatment with neutralizing antibodies and cytokine gene knockout mice. These studies are complicated by the fact that few, if any, cytokines are uniquely produced by CD8(+) T cells. Thus, the requirement for CD8(+) T cell- derived cytokines in resistance against most pathogens remains to be defined. Finally, recent studies of human CD8(+) T cells reveal the potential for novel effector mechanisms in resistance to infection. PMID- 10837061 TI - Glucocorticoids in T cell development and function*. AB - Glucocorticoids are small lipophilic compounds that mediate their many biological effects by binding an intracellular receptor (GR) that, in turn, translocates to the nucleus and directly or indirectly regulates gene transcription. Perhaps the most recognized biologic effect of glucocorticoids on peripheral T cells is immunosuppression, which is due to inhibition of expression of a wide variety of activationinduced gene products. Glucocorticoids have also been implicated in Th lineage development (favoring the generation of Th2 cells) and, by virtue of their downregulation of fasL expression, the inhibition of activation-induced T cell apoptosis. Glucocorticoids are also potent inducers of apoptosis, and even glucocorticoid concentrations achieved during a stress response can cause the death of CD4(+)CD8(+ )thymocytes. Perhaps surprisingly, thymic epithelial cells produce glucocorticoids, and based upon in vitro and in vivo studies of T cell development it has been proposed that these locally produced glucocorticoids participate in antigen-specific thymocyte development by inhibiting activation induced gene transcription and thus increasing the TCR signaling thresholds required to promote positive and negative selection. It is anticipated that studies in animals with tissue-specific GR-deficiency will further elucide how glucocorticoids affect T cell development and function. PMID- 10837062 TI - Molecular genetics of allergic diseases. AB - Allergic diseases affect approximately one third of the general population. This class of disease, characterized by elevated serum IgE levels and hypersensitivity to normally innocuous antigen, can manifest in practically any mucosal tissue or as a systemic response. A few examples of serious allergic diseases include asthma, dermatitis, bee sting allergy, food allergy, conjunctivitis, and severe systemic anaphylaxis. Taken together, allergic diseases constitute one of the major problems of modern day medicine. A considerable portion of the healthcare budget is expended in the treatment of allergic disease, and morbidity rates of inner city asthmatics are rising steadily. Due to the enormity of the problem, there has been a worldwide effort to identify factors that contribute to the etiology of allergic diseases. Epidemiologic studies of multigeneration families and large numbers of twins clearly indicate a strong genetic component to atopic diseases. At least two independently segregating diseasesusceptibility genes are thought to come together with environmental factors to result in allergic inflammation in a particular tissue. On the basis of the strong genetic studies, multiple groups have attempted to identify disease-susceptibility genes via either a candidate gene approach or by genome-wide scans. Both of these approaches have implicated multiple regions in the human and mouse genomes, which are currently being evaluated as harboring putative atopy genes. PMID- 10837063 TI - Immunology at the maternal-fetal interface: lessons for T cell tolerance and suppression. AB - Mammalian reproduction poses an immunological paradox because fetal alloantigens encoded by genes inherited from the father should provoke responses by maternal T cells leading to fetal loss. Current understanding of T cell immunobiology and the critical role of inflammatory processes during pregnancy is reviewed and discussed. Lessons derived from studies on the regulation of T cell responsiveness during mammalian gestation are considered in the wider context of T cell tolerance toward some microbial infections and tumors, avoidance of autoimmunity, and tissue allograft rejection. PMID- 10837064 TI - Regulation of B lymphocyte responses to foreign and self-antigens by the CD19/CD21 complex. AB - The membrane protein complex CD19/CD21 couples the innate immune recognition of microbial antigens by the complement system to the activation of B cells. CD21 binds the C3d fragment of activated C3 that becomes covalently attached to targets of complement activation, and CD19 co-stimulates signaling through the antigen receptor, membrane immunoglobulin. CD21 is also expressed by follicular dendritic cells and mediates the long-term retention of antigen that is required for the maintenance of memory B cells. Understanding of the biology of this receptor complex has been enriched by analyses of genetically modified mice; these analyses have uncovered roles not only in positive responses to foreign antigens, but also in the development of tolerance to self-antigens. Studies of signal transduction have begun to determine the basis for the coreceptor activities of CD19. The integration of innate and adaptive immune recognition at this molecular site on the B cell guides the appropriate selection of antigen by adaptive immunity and emphasizes the importance of this coreceptor complex. PMID- 10837065 TI - Regulatory T cells in autoimmmunity*. AB - Clonal deletion of autoreactive T cells in the thymus is not the sole mechanism for the induction of tolerance to self-antigens since partial depletion of peripheral CD4(+) T cells from neonatal and adult animals results in the development of organ-specific autoimmunity. Reconstitution of these immunodeficient animals with populations of regulatory CD4(+)T cells prevents the development of autoimmunity. The lineage of regulatory CD4(+) T cells is generated in the thymus and can be distinguished from effector cells by the expression of unique membrane antigens. The target antigens for these suppressor populations and their mechanisms of action remain poorly defined. Depletion of regulatory T cells may be useful in the induction of immunity to weak antigens, such as tumor-specific antigens. Conversely, enhancement of regulatory T cell function may be a useful adjunct to the therapy of autoimmune diseases and for prevention of allograft rejection. PMID- 10837066 TI - Signaling and transcription in T helper development. AB - The recognition of polarized T cell subsets defined by cytokine production was followed by a search to define the factors controlling this phenomenon. Suitable in vitro systems allowed the development of cytokine "recipes" that induced rapid polarization of naive T cells into Th1 or Th2 populations. The next phase of work over the past several years has begun to define the intracellular processes set into motion during Th1/Th2 development, particularly by the strongly polarizing cytokines IL-12 and IL-4. Although somewhat incomplete, what has emerged is a richly detailed tapestry of signaling and transcription, controlling an important T cell developmental switch. In addition several new mediators of control have emerged, including IL-18, the intriguing Th2-selective T1/ST2 product, and heterogeneity in dendritic cells capable of directing cytokine-independent Th development. PMID- 10837067 TI - The RAG proteins and V(D)J recombination: complexes, ends, and transposition. AB - V(D)J recombination proceeds through a series of protein:DNA complexes mediated in part by the RAG1 and RAG2 proteins. These proteins are responsible for sequence-specific DNA recognition and DNA cleavage, and they appear to perform multiple postcleavage roles in the reaction as well. Here we review the interaction of the RAG proteins with DNA, the chemistry of the cleavage reaction, and the higher order complexes in which these events take place. We also discuss postcleavage functions of the RAG proteins, including recent evidence indicating that they initiate the process of coding end processing by nicking hairpin DNA termini. Finally, we discuss the evolutionary and functional implications of the finding that RAG1 and RAG2 constitute a transposase, and we consider RAG protein biochemistry in the context of several bacterial transposition systems. This suggests a model of the RAG protein active site in which two divalent metal ions serve alternating and opposite roles as activators of attacking hydroxyl groups and stabilizers of oxyanion leaving groups. PMID- 10837068 TI - The role of the thymus in immune reconstitution in aging, bone marrow transplantation, and HIV-1 infection. AB - The human thymus is a complex chimeric organ comprised of central (thymic epithelial space) and peripheral (perivascular space) components that functions well into adult life to produce naive T lymphocytes. Recent advances in identifying thymic emigrants and development of safe methods to study thymic function in vivo in adults have provided new opportunities to understand the role that the human thymus plays in immune reconstitution in aging, in bone marrow transplantation, and in HIV-1 infection. The emerging concept is that there are age-dependent contributions of thymic emigrants and proliferation of postthymic T cells to maintain the peripheral T cell pool and to contribute to T cell regeneration, with the thymus contributing more at younger ages and peripheral T cell expansion contributing more in older subjects. New studies have revealed a dynamic interplay between postnatal thymus output and peripheral T cell pool proliferation, which play important roles in determining the nature of immune reconstitution in congenital immunodeficiency diseases, in bone marrow transplantation, and in HIV-1 infection. In this paper, we review recent data on human postnatal thymus function that, taken together, support the notion that the human thymus is functional well into the sixth decade and plays a role throughout life to optimize human immune system function. PMID- 10837069 TI - Accessing complexity: the dynamics of virus-specific T cell responses. AB - The cellular dynamics of the immune system are complex and difficult to measure. Access to this problematic area has been greatly enhanced by the recent development of tetrameric complexes of MHC class I glycoprotein + peptide (tetramers) for the direct staining of freshly isolated, antigen-specific CD8(+ )T cells. Analysis to date with both naturally acquired and experimentally induced infections has established that the numbers of virus-specific CD8(+) T cells present during both the acute and memory phases of the host response are more than tenfold in excess of previously suspected values. The levels are such that the virus-specific CD8(+) set is readily detected in the human peripheral blood lymphocyte compartment, particularly during persistent infections. Experimentally, it is now possible to measure the extent of cycling for tetramer (+)CD8(+) T cells during the acute and memory phases of the host response to viruses. Dissection of the phenotypic, functional, and molecular diversity of CD8(+) T cell populations has been greatly facilitated. It is hoped it will also soon be possible to analyze CD4(+) T cell populations in this way. Though these are early days and there is an enormous amount to be done, our perceptions of the shape of virus-specific cell-mediated immunity are changing rapidly. PMID- 10837070 TI - The role of chemokine receptors in primary, effector, and memory immune responses. AB - The immune system is composed of single cells, and its function is entirely dependent on the capacity of these cells to traffic, localize within tissues, and interact with each other in a precisely coordinated fashion. There is growing evidence that the large families of chemokines and chemokine receptors provide a flexible code for regulating cell traffic and positioning in both homeostatic and inflammatory conditions. The regulation of chemokine receptor expression during development and following cell activation explains the complex migratory pathways taken by dendritic cells, T and B lymphocytes, providing new insights into the mechanisms that control priming, effector function, and memory responses. PMID- 10837071 TI - Phosphorylation meets ubiquitination: the control of NF-[kappa]B activity. AB - NF-kappaB (nuclear factor-kappaB) is a collective name for inducible dimeric transcription factors composed of members of the Rel family of DNA-binding proteins that recognize a common sequence motif. NF-kappaB is found in essentially all cell types and is involved in activation of an exceptionally large number of genes in response to infections, inflammation, and other stressful situations requiring rapid reprogramming of gene expression. NF-kappaB is normally sequestered in the cytoplasm of nonstimulated cells and consequently must be translocated into the nucleus to function. The subcellular location of NF kappaB is controlled by a family of inhibitory proteins, IkappaBs, which bind NF kappaB and mask its nuclear localization signal, thereby preventing nuclear uptake. Exposure of cells to a variety of extracellular stimuli leads to the rapid phosphorylation, ubiquitination, and ultimately proteolytic degradation of IkappaB, which frees NF-kappaB to translocate to the nucleus where it regulates gene transcription. NF-kappaB activation represents a paradigm for controlling the function of a regulatory protein via ubiquitination-dependent proteolysis, as an integral part of a phosphorylationbased signaling cascade. Recently, considerable progress has been made in understanding the details of the signaling pathways that regulate NF-kappaB activity, particularly those responding to the proinflammatory cytokines tumor necrosis factor-alpha and interleukin-1. The multisubunit IkappaB kinase (IKK) responsible for inducible IkappaB phosphorylation is the point of convergence for most NF-kappaB-activating stimuli. IKK contains two catalytic subunits, IKKalpha and IKKbeta, both of which are able to correctly phosphorylate IkappaB. Gene knockout studies have shed light on the very different physiological functions of IKKalpha and IKKbeta. After phosphorylation, the IKK phosphoacceptor sites on IkappaB serve as an essential part of a specific recognition site for E3RS(IkappaB/beta-TrCP), an SCF type E3 ubiquitin ligase, thereby explaining how IKK controls IkappaB ubiquitination and degradation. A variety of other signaling events, including phosphorylation of NF-kappaB, hyperphosphorylation of IKK, induction of IkappaB synthesis, and the processing of NF-kappaB precursors, provide additional mechanisms that modulate the level and duration of NF-kappaB activity. PMID- 10837072 TI - Reservoirs for HIV-1: mechanisms for viral persistence in the presence of antiviral immune responses and antiretroviral therapy. AB - The success of combination antiretroviral therapy for HIV-1 infection has generated interest in mechanisms by which the virus can persist in the body despite the presence of drugs that effectively inhibit key steps in the virus life cycle. It is becoming clear that viral reservoirs established early in the infection not only prevent sterilizing immunity but also represent a major obstacle to curing the infection with the potent antiretroviral drugs currently in use. Mechanisms of viral persistence are best considered in the context of the dynamics of viral replication in vivo. Virus production in infected individuals is largely the result of a dynamic process involving continuous rounds of de novo infection of and replication in activated CD4(+) T cells with rapid turnover of both free virus and virus-producing cells. This process is largely, but not completely, interrupted by effective antiretroviral therapy. After a few months of therapy, plasma virus levels become undetectable in many patients. Analysis of viral decay rates initially suggested that eradication of the infection might be possible. However, there are several potential cellular and anatomical reservoirs for HIV-1 that may contribute to long-term persistence of HIV-1. These include infected cell in the central nervous system and the male urogenital tract. However, the most worrisome reservoir consists of latently infected resting memory CD4(+) T cells carrying integrated HIV-1 DNA. Definitive demonstration of the presence of this form of latency required development of methods for isolating extremely pure populations of resting CD4(+) T cells and for demonstrating that a small fraction of these cells contain integrated HIV-1 DNA that is competent for replication if the cells undergo antigen-driven activation. Most of the latent virus in resting CD4(+) T cells is found in cells of the memory phenotype. The half-life of this latent reservoir is extremely long (44 months). At this rate, eradication of this reservoir would require over 60 years of treatment. Thus, latently infected resting CD4(+) T cells provide a mechanism for life-long persistence of replication-competent forms of HIV-1, rendering unrealistic hopes of virus eradication with current antiretroviral regimens. The extraordinary stability of the reservoir may reflect gradual reseeding by a very low level of ongoing viral replication and/or mechanisms that contribute to the intrinsic stability of the memory T cell compartment. Given the substantial long term toxicities of current combination therapy regimens, novel approaches to eradicating this latent reservoir are urgently needed. PMID- 10837073 TI - Regulation of antibody responses via antibodies, complement, and Fc receptors. AB - Antibodies can completely suppress or enhance the antibody response to their specific antigen by several hundredfold. Immunoglobulin M (IgM) enhances antibody responses via the complement system, and complement activation by IgM probably starts the chain of events leading to antibody responses to suboptimal antigen doses. IgG can enhance primary antibody responses in the absence of the complement system and seems to be dependent on Fc receptors for IgG (FcgammaRs). IgE enhances antibody responses via the low-affinity receptor for IgE (FcepsilonRII/CD23). The precise effector mechanisms that cause enhancement are not known, but direct B-cell signaling, antigen presentation, and increased follicular localization are all possibilities. IgG, IgE, and IgM may also suppress antibody responses when used in certain immunization regimes, and it seems reasonable that an important mechanism behind suppression is the masking of antigenic epitopes by antibodies. In addition, FcgammaRIIB, which contains a cytoplasmic inhibitory motif, acts as a negative regulator of antibody responses. This receptor, however, may prevent the antibody responses from exceeding a certain level rather than causing complete suppression. PMID- 10837074 TI - Multiple roles for the major histocompatibility complex class I- related receptor FcRn. AB - Multiple functions have recently been identified for the neonatal Fc receptor FcRn. In addition, a human homolog of the rodent forms of FcRn has been identified and characterized. This major histocompatibility complex class I related receptor plays a role in the passive delivery of immunoglobulin (Ig)Gs from mother to young and the regulation of serum IgG levels. In addition, FcRn expression in tissues such as liver, mammary gland, and adult intestine suggests that it may modulate IgG transport at these sites. These diverse functions are apparently brought about by the ability of FcRn to bind IgGs and transport them within and across cells. However, the molecular details as to how FcRn traffics within cells have yet to be fully understood, although in vitro systems have been developed for this purpose. The molecular nature of the FcRn-IgG interaction has been studied extensively and encompasses residues located at the CH2-CH3 domain interface of the Fc region of IgG. These Fc amino acids are highly conserved in rodents and man and interact with residues primarily located on the alpha2 domain of FcRn. Thus, it is now possible to engineer IgGs with altered affinities for FcRn, and this has relevance to the modulation of IgG serum half-life and maternofetal IgG transport for therapeutic applications. PMID- 10837075 TI - Immunobiology of dendritic cells. AB - Dendritic cells (DCs) are antigen-presenting cells with a unique ability to induce primary immune responses. DCs capture and transfer information from the outside world to the cells of the adaptive immune system. DCs are not only critical for the induction of primary immune responses, but may also be important for the induction of immunological tolerance, as well as for the regulation of the type of T cell-mediated immune response. Although our understanding of DC biology is still in its infancy, we are now beginning to use DC-based immunotherapy protocols to elicit immunity against cancer and infectious diseases. PMID- 10837076 TI - An address system in the vasculature of normal tissues and tumors. AB - The vasculature of individual tissues is highly specialized. The endothelium in lymphoid tissues expresses tissue-specific receptors for lymphocyte homing, and recent work utilizing phage homing has revealed an unprecedented degree of specialization in the vasculature of other normal tissues. In vivo screening of libraries of phage that displace random peptide sequences on their surfaces has yielded specific homing peptides for a large number of normal tissues. The tissue specific endothelial molecules to which the phage peptides home may serve as receptors for metastasizing malignant cells. Probing of tumor vasculature has yielded peptides that home to endothelial receptors expressed selectively in angiogenic neovasculature. These receptors, and those specific for the vasculature of individual normal tissues, are likely to be useful in targeting therapies to specific sites. PMID- 10837077 TI - Genomic views of the immune system*. AB - Genomic-scale experimentation aims to view biological processes as a whole, yet with molecular precision. Using massively parallel DNA microarray technology, the mRNA expression of tens of thousands of genes can be measured simultaneously. Mathematical distillation of this flood of gene expression data reveals a deep molecular and biological logic underlying gene expression programs during cellular differentiation and activation. Genes that encode components of the same multi-subunit protein complex are often coordinately regulated. Coordinate regulation is also observed among genes whose products function in a common differentiation program or in the same physiological response pathway. Recent application of gene expression profiling to the immune system has shown that lymphocyte differentiation and activation are accompanied by changes of hundreds of genes in parallel. The databases of gene expression emerging from these studies of normal immune responses will be used to interpret the pathological changes in gene expression that accompany autoimmunity, immune deficiencies, and cancers of immune cells. PMID- 10837078 TI - Viral subversion of the immune system. AB - This review describes the diverse array of pathways and molecular targets that are used by viruses to elude immune detection and destruction. These include targeting of pathways for major histocompatibility complex-restricted antigen presentation, apoptosis, cytokine-mediated signaling, and humoral immune responses. The continuous interactions between host and pathogens during their coevolution have shaped the immune system, but also the counter measures used by pathogens. Further study of their interactions should improve our ability to manipulate and exploit the various pathogens. PMID- 10837079 TI - DNA vaccines: immunology, application, and optimization*. AB - The development and widespread use of vaccines against infectious agents have been a great triumph of medical science. One reason for the success of currently available vaccines is that they are capable of inducing long-lived antibody responses, which are the principal agents of immune protection against most viruses and bacteria. Despite these successes, vaccination against intracellular organisms that require cell-mediated immunity, such as the agents of tuberculosis, malaria, leishmaniasis, and human immunodeficiency virus infection, are either not available or not uniformly effective. Owing to the substantial morbidity and mortality associated with these diseases worldwide, an understanding of the mechanisms involved in generating long-lived cellular immune responses has tremendous practical importance. For these reasons, a new form of vaccination, using DNA that contains the gene for the antigen of interest, is under intensive investigation, because it can engender both humoral and cellular immune responses. This review focuses on the mechanisms by which DNA vaccines elicit immune responses. In addition, a list of potential applications in a variety of preclinical models is provided. PMID- 10837080 TI - [gamma][delta] cells: a right time and a right place for a conserved third way of protection. AB - The tripartite subdivision of lymphocytes into B cells, alphabeta T cells, and gammadelta cells has been conserved seemingly since the emergence of jawed vertebrates, more than 450 million years ago. Yet, while we understand much about B cells and alphabeta T cells, we lack a compelling explanation for the evolutionary conservation of gammadelta cells. Such an explanation may soon be forthcoming as advances in unraveling the biochemistry of gammadelta cell interactions are reconciled with the abnormal phenotypes of gammadelta-deficient mice and with the striking differences in gammadelta cell activities in different strains and species. In this review, the properties of gammadelta cells form a basis for understanding gammadelta cell interactions with antigens and other cells that in turn form a basis for understanding immunoprotective and regulatory functions of gammadelta cells in vivo. We conclude by considering which gammadelta cell functions may be most critical. PMID- 10837081 TI - New rulings on the Cetus patent for taq polymerase. PMID- 10837082 TI - Metabolic pathway for leucovorin. PMID- 10837083 TI - The promise and the dilemma of the new millennium. PMID- 10837084 TI - The political voice of molecular pathologists. PMID- 10837085 TI - Blood screening by nucleic acid amplification technology: current issues, future challenges. AB - BACKGROUND: Nucleic acid amplification technology (NAT) is presently being evaluated in US clinical trials to determine the safety and efficacy of mini-pool testing for human immunodeficiency virus (HIV) and hepatitis C virus (HCV) RNA in the blood-donor population. Although the risk for transfusion-transmitted HIV and HCV infection is extremely low, there is still a small chance that blood donated by infected individuals before seroconversion can escape detection by current antibody-based assays. METHODS: This report describes the amplification technologies being used and reviews several issues surrounding NAT-based blood screening. The performance features of NAT and current enzyme immunoassay technologies are compared, and the benefits of NAT in reducing transfusion transmitted infections are discussed. CONCLUSIONS: The current US clinical trials of mini-pool NAT testing for HIV and HCV RNA have successfully identified preseroconversion infectious blood units. Although the current NAT-based screening systems are semiautomated, mini-pool testing represents an unprecedented innovation among government and nongovernment agencies in the highly regulated blood transfusion industry. Despite cost-effectiveness issues, based on the public perception of infectious diseases acquired through blood transfusion, NAT-based screening of the blood supply is expected to become a standard in transfusion medicine. PMID- 10837086 TI - Identification of a high frequency of chromosomal rearrangements in the centromeric regions of prostate cancer cell lines by sequential giemsa banding and spectral karyotyping. AB - BACKGROUND: Currently, prostate cancer (CaP) cytogenetics is not well defined, largely because of technical difficulties in obtaining primary tumor metaphases. METHODS AND RESULTS: We examined three CaP cell lines (LNCaP, DU145, PC-3) using sequential Giemsa banding and spectral karyotyping (SKY) to search for a common structural aberration or translocation breakpoint. No consistent rearrangement common to all three cell lines was detected. A clustering of centromeric translocation breakpoints was detected in chromosomes 4, 5, 6, 8, 11, 12, 14, and 15 in DU145 and PC-3. Both these lines were found to have karyotypes with a greater level of complexity than LNCaP. CONCLUSION: The large number of structural aberrations present in DU145 and PC-3 implicate an underlying chromosomal instability and subsequent accumulation of cytogenetic alterations that confer a selective growth advantage. The high frequency of centromeric rearrangements in these lines indicates a potential role for mitotic irregularities associated with the centromere in CaP tumorigenesis. PMID- 10837087 TI - Detection of circulating epithelial cells after surgery for benign breast disease. AB - BACKGROUND: Cytokeratins are predominantly expressed in epithelial cells and their malignant counterparts. Ultrasensitive methods for cytokeratin messenger RNAs (mRNAs) can detect rare circulating tumor cells consistent with hematogenous dissemination in epithelial-derived malignancies, including breast carcinomas. Intraoperative tumor-cell shedding may contribute to this process; this hypothesis is based on the assumption that only tumor cells can be mobilized during surgical manipulation. METHODS AND RESULTS: The present study addresses this issue by using cytokeratin 19 mRNA detection by reverse transcription polymerase chain reaction (RT-PCR) in preoperative and postoperative blood samples from 54 patients undergoing excisional biopsy for benign breast disease; 22 healthy volunteers represented the control group. No cytokeratin RT-PCR positivity was found in the control or preoperative samples. Cytokeratin RT-PCR positivity was found in 21 postoperative samples (39%). CONCLUSIONS: This finding shows that benign epithelial cells can be mobilized during breast surgery; this effect of surgical manipulation warrants caution in the interpretation of RT-PCR positivity for cytokeratin mRNA in the peripheral blood of patients undergoing surgery for breast cancer. PMID- 10837088 TI - Mitochondrial ATP synthase 6 as an endogenous control in the quantitative RT-PCR analysis of clinical cancer samples. AB - BACKGROUND: Real-time polymerase chain reaction (PCR) is a powerful new technique in the evolution of quantitative reverse transcription-PCR assays. With the increased sensitivity and resolution of real-time techniques, the requirements for constitutive expression of endogenous controls have become increasingly stringent. METHODS AND RESULTS: We compare the expression of the mitochondrial gene, adenosine triphosphate synthase 6 (ATPsy6), to the expression of other routinely used endogenous control genes (e.g., beta-actin, glyceraldehyde-3 phosphate dehydrogenase [GAPDH], ribosomal RNA 18S [18S rRNA], and cyclophilin). In a diverse assortment of tissues and across a wide range of disease stages, ATPsy6 shows a relative steady state of expression compared with other endogenous controls. ATPsy6 gene expression has been used as an endogenous control in a quantitative real-time PCR assay designed to evaluate the expression of potential cancer diagnostic leads across a diverse tissue panel. CONCLUSION: Mitochondrial ATPsy6 serves as a good endogenous control to measure target gene expression independent of the tissue- or disease-specific variation inherent with many housekeeping genes. PMID- 10837089 TI - Analysis of CYP21 coding polymorphisms in three ethnic populations: further evidence of nonamplifying CYP21 alleles among whites. AB - BACKGROUND: Adrenal steroid 21-hydroxylase is essential for the synthesis of both mineralocorticoids and glucocorticoids. The gene for this enzyme, CYP21, contains several frequent coding polymorphisms. Because of its essential function in steroid synthesis, polymorphisms in this enzyme might influence a variety of disease processes. However, before disease-association studies are performed, it is important to understand the frequency of these polymorphisms among normal individuals. METHODS: Using polymerase chain reaction (PCR) with restriction enzyme digestion or size length polymorphism analysis, we measured the frequencies of the +Leu(10), Arg102Lys, and Ser268Thr polymorphisms in CYP21 in healthy whites, blacks, and Indian Americans. The subjects were all young female college students participating in a study of relative risks for cardiovascular disease in these populations. RESULTS: The frequency of each polymorphism among whites, blacks, and Indian Americans were as follows: +Leu(10), 0.55, 0.96, 0.75; Arg102, 0.63, 0.97, 0.82; and Ser268, 0.92, 0.68, 0.79, respectively. With the exception of the frequencies of the Ser268Thr polymorphism among blacks and Indian Americans, there were significantly different frequencies of each polymorphism among all groups (P<.05). Among whites, the distribution of genotypes for the +Leu(10) and Arg102Lys polymorphisms deviated significantly from expected Hardy-Weinberg values because of an excess of homozygotes. CONCLUSIONS: Among the ethnic groups, there are statistically significant differences in the frequencies of these common coding polymorphisms in CYP21 that need to be considered in disease-association studies. Deviation from Hardy Weinberg distributions might be explained by allelic dropout during PCR, a phenomenon previously reported at this locus. PMID- 10837090 TI - Fatal pulmonary embolism: a study of genetic and acquired factors. AB - BACKGROUND: Investigators speculate that hereditary thrombotic disease coupled with acquired factors account for a large number of pulmonary thromboemboli. Clinical correlation between genetic and acquired factors with fatal pulmonary thromboemboli has not been extensively studied. METHODS: Archival autopsy material was obtained from patients who died of or with pulmonary emboli for whom confirmed autopsy results were available. Polymerase chain reaction-restriction fragment length polymorphism analysis was performed for factor V Leiden and factor II/20210A allele. Retrospective chart review was performed to determine the presence or absence of acquired factors that can predispose to pulmonary thromboemboli. RESULTS: Two of 36 patients (5.5%) were heterozygous for factor V Leiden. No patients had detected abnormalities for factor II/20210A allele. Eight patients (22.2%) had a malignancy present, one of whom was heterozygous for factor V Leiden. Fourteen patients (38.8%) had recent major surgery or were immobilized. CONCLUSIONS: The incidence of factor V Leiden and factor II/20210A allele in patients with fatal pulmonary thromboemboli is not greater than published results of the incidence of these factors in the general population. Recognized acquired factors such as malignancy, recent surgery, and immobilization are frequently present in these patients. Our results suggest that genetic profiling of thrombotic disease will not replace clinical vigilance in reducing the risk for death from pulmonary thromboemboli. PMID- 10837091 TI - Prevalence of methylenetetrahydrofolate reductase mutations in patients with venous thrombosis. AB - BACKGROUND: The objectives of this study are to examine the prevalence of combined methylenetetrahydrofolate reductase (MTHFR) 677C-->T and 1298A-->C mutations in patients with venous thrombosis (VT) and healthy volunteers and to determine if these mutations are in Hardy-Weinberg equilibrium. METHODS AND RESULTS: Sixty-five patients with VT and 64 healthy volunteers were assessed for MTHFR 677T and 1298C alleles using polymerase chain reaction and restriction fragment length polymorphism. Observed MTHFR genotype frequencies were compared with expected genotype combinations, and their odds ratios were determined. MTHFR allele frequency did not differ between VT and control groups; however, differences were observed for MTHFR genotype distribution. MTHFR 677T and 1298C alleles occurred in cis in our population, and therefore mutation crossover has occurred. There was deviation from the Hardy-Weinberg equilibrium for combined MTHFR genotypes, although this may at least partly be attributable to linkage disequilibrium. MTHFR 677CT/1298CC and 677TT/1298CC genotypes (P<.05) were not observed in either group. CONCLUSIONS: The absence of MTHFR 677CT/1298CC and 677TT/1298CC genotypes in both groups suggests that certain MTHFR genotypes may carry a selective advantage. Our discovery of a substantial number of MTHFR mutations in cis configuration suggests that any MTHFR allele linkage disequilibrium present is incomplete. PMID- 10837092 TI - Correct heteroduplex formation for mutation detection analysis. AB - BACKGROUND: The majority of mutation detection methods for unknown mutations are polymerase chain reaction (PCR)-based methods dependent on the formation of heteroduplexes between wild-type and mutant strands of DNA. METHODS AND RESULTS: This report discusses the difficulties associated with forming heteroduplexes with a large DNA fragment and the implications for subsequent mutation detection by the chemical cleavage of mismatch technique and other methods reliant on heteroduplex formation. It was found that the size and sequence context of the fragment being investigated inhibited correct heteroduplex formation. The problem was overcome by dividing the sequence into two overlapping fragments. CONCLUSIONS: Early identification of this problem in other fragments will help with the rapid optimization of PCR-based mutation detection methods. PMID- 10837094 TI - Clinical news update PMID- 10837093 TI - A simple PCR test to detect the common 35delG mutation in the connexin 26 gene. AB - BACKGROUND: The most common form of nonsyndromic neurosensory autosomal recessive deafness, DFNB1, is caused by mutations in the connexin 26 gene (GJB2) on chromosome 13. One mutation, in which one guanosine (G) residue is deleted from a run of 6 Gs (35delG), is found in 40% to 70% of DFNB1 cases and has an expected population frequency of one in 40 to one in 100. METHODS AND RESULTS: Polymerase chain reaction (PCR)-based tests for the 35delG mutation were developed. They are based on mismatched PCR primers that produce novel EcoRII or DdeI restriction enzyme sites depending on the number of Gs at the 35delG locus. An EcoRII site is generated in the wild-type sequence (6 Gs), but not when the 35delG mutation is present. Alternatively, a DdeI site can be generated so that this enzyme cuts the PCR product when the 35delG mutation is present, but not the wild-type sequence. CONCLUSIONS: These tests enable a quick and reliable screen for the common 35delG mutation. PMID- 10837095 TI - The epidemiology of delirium: a review of studies and methodological issues. AB - Information about the epidemiology of delirium has grown tremendously over the past 2 decades. However, methodological challenges have made this area of research difficult to study. Most studies have focused on the prevalence, risk factors, and course of delirium in elderly populations and acute medical and surgical settings. Growing evidence is showing that delirium is a neuropsychiatric syndrome that deserves more research and clinical attention. This article summarizes the current literature on the epidemiology of delirium and discusses some of the important methodological issues in conducting and interpreting research in this area. PMID- 10837096 TI - Motoric subtypes of delirium. AB - Delirium is a common neuropsychiatric disorder with wide ranging symptoms and significant morbid impact. Disturbances of motor behavior are an important feature of delirium and form the basis for the most commonly studied clinical subtype. This article reviews the relevance of motoric disturbance to delirium phenomenology and discusses possible neurobiological causes for different presentations of motor behavior in delirium. Evidence is presented to support the usefulness of using motorically defined subtypes based on identified differences according to underlying origins, pathophysiologies, responsiveness to therapy and natural course. Methodological issues relating to motoric subtype studies are addressed and suggestions for future research are made. PMID- 10837097 TI - EEG in delirium. AB - Electroencephalography (EEG) is an inexpensive and noninvasive probe of functional brain activity that is one of the few clinically accessible measures capable of detecting changes in delirium. In EEG characteristics of delirium include slowing or dropout of the posterior dominant rhythm, generalized theta or delta slow-wave activity, poor organization of the background rhythm, and loss of reactivity of the EEG to eye opening and closing. These are paralleled by the quantitative EEG (QEEG) findings of increased absolute and relative slow-wave (theta and delta) power, reduced ratio of fast-to-slow band power, reduced mean frequency, and reduced occipital peak frequency. In alcohol and sedative withdrawal, EEG findings may include attenuation of voltage and prominence of beta activity. Specificity of EEG and QEEG findings in delirium has been questioned, particularly vis-'a-vis EEG changes in normal aging and in dementia. As noted in the text, however, specificity can be increased by the use of careful data acquisition and skillful interpretation. In the future, it is expected that QEEG methods will largely replace EEG in application to delirium. PMID- 10837098 TI - The interaction of delirium and seizures. AB - The induction of a delirium by medical illness, somatic treatments, or experimental drugs occasionally relieves psychotic, excited, and manic states. An induced delirium is a feature of modern electroconvulsive therapy (ECT), and was a feature of insulin coma therapy and psychosurgery. Case material explores the relationship between psychosis, mania, seizures, and electroencephalogram. From our understanding of the mechanism of ECT in relieving intractable status epilepticus, we suggest a hypothesis for the beneficial interaction between delirium and ECT. PMID- 10837099 TI - Neuroimaging in delirium and related conditions. AB - Delirium is one of the most fascinating and poorly understood syndromes in medicine. To a large extent, attempts to study the pathophysiology of delirium have been hampered by the many different types of delirium and their variable symptom expression. The emergence of sophisticated brain imaging methodologies has made it possible to move beyond diagnostic considerations and investigate the neurobiology of specific symptom clusters observed in delirium and related conditions. In this review, neuroimaging findings of relevance to delirium are interpreted in relation to phenomenologically similar symptom states as well as clinical diagnoses. A promising approach in this regard is to combine neuroimaging techniques with symptom-provoking pharmacologic challenge paradigms. Such symptom-oriented neuroimaging studies hold particular promise for improving our understanding of the pathophysiology of delirium and its treatment. PMID- 10837100 TI - Drug-induced delirium. AB - Drug-induced delirium (DID) is a common and debilitating clinical problem. Identifying and treating the causes of DID is becoming easier as knowledge of the pharmacodynamic and pharmacokinetic fates of drugs increases. This article reviews basic pharmacological mechanisms and risks of DID, and ways to manage these risks. PMID- 10837101 TI - Serotonin and amino acids: partners in delirium pathophysiology? AB - Delirium may be the result of dysfunction of multiple interacting neurotransmitter systems. Changes in the levels of various amino acids being precursors of cerebral neurotransmitters may affect their function and, thus, contribute to the development of delirium. Serotonin is one of the neurotransmitters that may play an important role in medical and surgical delirium. Normal serotonin synthesis and release in the human brain is, among others, dependent on the availability of its precursor tryptophan (Trp) from blood. The essential amino acid Trp competes with the other large neutral amino acids (LNAA) tyrosine, phenylalanine, valine, leucine, and isoleucine for transport across the blood-brain barrier. This competition determines its uptake into the brain, represented by the ratio of the plasma level of Trp to the sum of the other LNAA. The plasma ratio of Trp/LNAA, plasma level of Trp, and serotonin in plasma and platelets have been used as indirect peripheral measures for central serotonergic functioning. Both increased and decreased serotonergic activity have been associated with delirium. Serotonin agonists can induce psychosis, both elevated Trp availability and increased cerebral serotonin have been associated with hepatic encephalopathy, and excess serotonergic brain activity has been related to the development of the serotonin syndrome of which delirium is a main symptom. On the other hand, alcohol withdrawal delirium, delirium in levodopa-treated Parkinson patients, and postoperative delirium have been related to reduce cerebral Trp availability from plasma suggesting diminished serotonergic function. Rick factors for delirium such as severe illness, surgery, and trauma can induce immune activation and a physical stress response comprising increased activity of the limbic-hypothalamic-pituitary adrenocortical axis, the occurrence of a low T3 syndrome, and, possibly, changes in the permeability of the blood-brain barrier. There are indications that these changes have their effect on plasma amino acid concentrations, e.g., Trp, and multiple cerebral neurotransmitters, including serotonin. This stress response may be different depending on the stage of illness being acute or chronic. It will require further study to determine the complex influence of the stress response and immune activation on plasma amino acids, neurotransmitter function and the development of delirium, especially in the more vulnerable older patients. PMID- 10837102 TI - Is there a final common neural pathway in delirium? Focus on acetylcholine and dopamine. AB - This article reviews the literature relevant to improving our understanding of the neural underpinnings of delirium. That the characteristic symptoms of delirium occur as a result of a wide diversity of causes supports the concept of a ""final common pathway. " What constitutes this may involve certain brain regions or circuits and certain neurotransmitters. Neuroanatomical data derived from neuroimaging and lesion reports suggest the importance of pathways in prefrontal cortex, thalamus, fusiform cortex, posterior parietal cortex, and basal ganglia. Neurotransmitters most implicated in delirium that could be candidates to mediate the characteristic symptoms of delirium, as well as the electroencephalogram changes, are acetylcholine and dopamine. Acetylcholine deficiency and dopamine excess---absolute and/or relative to each other---appear to be critical in the final common pathway. These neurotransmitters affect each other, depending on the receptor subtype, and their receptor distribution among layers of cortex in areas such as prefrontal cortex and temporal lobe suggests that cholinergic and dopaminergic neurons could interact with each other during delirium. Electroconvulsive therapy is described as a special situation in which excess dopamine and delirium may have a therapeutic effect on depression recovery, in contrast with the usual association of delirium with negative effects. PMID- 10837103 TI - Serum anticholinergic activity levels and delirium in the elderly. AB - This article will briefly review the clinical studies focusing on measurement of serum levels of anticholinergic activity in delirious states. Three experimental approaches have been taken. First, to identify medications currently prescribed that have subtle anticholinergic effects. The current "list" includes 48 commonly prescribed medications. Second, to associate serum anticholinergic activity with delirium in various clinical states including postcardiotomy delirium, postelectroconvulsive delirium, delirious elderly medical inpatients, and nursing home patients. Third, to intervene in patients with elevated anticholinergic activity by reducing known anticholinergics and correlating this reduction with clinical measures of cognition and delirium. Our most recent data investigate the impact of anticholinergics on demented patients. Prevalence of delirium was significantly higher in patients receiving larger numbers of anticholinergics. PMID- 10837104 TI - Throughout Europe and North America young people at the interface of criminal justice systems and mental health services risk double jeopardy for social exclusion alienation and stigmatization. PMID- 10837105 TI - Mental health needs and services for severely troubled and troubling young people including young offenders in an N.H.S. region. AB - Penal, social services, special education and health agencies in one National Health Service Region were screened for severely troubled and troublesome young people in need of mental health services. One hundred and sixteen detailed assessments were undertaken on young people: 57 in "penal" and 59 in "welfare" establishments. Little difference was found between those in these two types of setting except that the "penal group" were much more likely to have high levels of violent behaviour and to have had more changes of placement. The needs for mental health care greatly outstripped supply. As part of this overall neglect, 11 of 15 young people with serious mental illnesses and all 13 who had suffered sexual abuse in the sample were not receiving appropriate treatment. A tentative estimate of the size of the problem in the region yielded a rate of around 11.4 per million with very severe disorder but this is probably an under estimate. The effectiveness of treatment for the problems of these young people is discussed and a possible structure for a service is explored. PMID- 10837106 TI - Forensic assessment of juvenile delinquents: prevalence of psychopathology and decision-making at court in the netherlands. AB - Dutch juvenile criminal law, concerning minors between 12 and 18 years old, is marked by its pedagogical character. In cases of serious psychosocial problems or psychiatric disorders, the juvenile court may request a forensic examination in order to be able to impose a measure of restraint. This study was aimed at investigating to what extent the juvenile criminal law achieves the aim enhancing professional help in those cases for which such assistance is indicated. One hundred and eight juveniles who were brought before the court were assessed multidisciplinarily. Sixty-five per cent of the youngsters were qualified for 'definite caseness'. For less than half of them a forensic assessment was ordered by the court. In conclusion, young delinquents should be screened for psychiatric disorders more adequately as soon as they are taken into custody. PMID- 10837107 TI - A descriptive survey of Flemish delinquent adolescents. AB - Previous research has shown that delinquent adolescents are characterized by a number of psychiatric problems. Most of these assessments, however, have been conducted on incarcerated adolescents. By means of semi-structured assessment (Child Assessment Schedule) and self-report measures, we assessed psychiatric status in a group of 72 delinquent adolescents, adjudicated before the Juvenile Court of Antwerp, Belgium. A significant difference was found between ethnic groups on self-report scores. Seventy per cent of subjects received at least one psychiatric diagnosis. The most prevalent diagnoses were conduct disorder (CD), substance abuse disorder and attention deficit hyperactivity disorder (ADHD). Less prevalent were post traumatic stress disorder (PTSD) and major depression (MD). As we found that an unselected sample of delinquent adolescents show a great number of psychiatric disorders, we suggest that psychiatric screening of delinquent adolescents should be done systematically. The predictive value of psychiatric assessment and subsequent treatment in delinquent adolescents should be investigated by future, prospective research. PMID- 10837108 TI - Girls in detention: what are their characteristics? A project to explore and document the character of this target group and the significant ways in which it differs from one consisting of boys. AB - In this article, the differences between male and female juvenile delinquency are discussed, from a behavioural and psychiatric perspective. At first the existence and nature of female juvenile delinquency is described. Further a, mostly literature-based, analysis of gender-related differences in juvenile delinquency and psychopathology is presented. Finally the results are given of a dossier research project into the behavioural aspects of female juvenile delinquency. Girls commit fewer and less serious offences. Girls in a detention centre are younger than boys in the same centre. Few Turkish and Moroccan girls are seen. The majority of girls are placed in a detention centre by a Civil Court Order. A relatively high percentage of retarded girls was found. Multiple sexual contacts from an early age, substance abuse, running away from home and truancy are risk behaviours in the development of female juvenile delinquency. In the majority of cases there was no record of previous contacts with the law. It seems as if there are gender-related factors according to the development of female juvenile delinquency. Answers on age-differences, early sexual development, the relation between behaviour and psychiatric diagnosis, the role of the risk behaviours in the development of the girls, intelligence and the reasons for placing girls in a detention centre should throw light on the development process of female juvenile delinquents. PMID- 10837109 TI - Male adolescent sex offenders against children: similarities and differences between those offenders with and those without a history of sexual abuse. AB - This study compares a defined sample of male adolescents (n = 36) with regard to their personal development, family characteristics and the types of offence they committed. Adolescent sex offenders with (n = 16) and without (n = 20) a history of sexual abuse who had offended against children were investigated during ongoing criminal proceedings by means of questionnaires and intelligence tests. The most important characteristic that distinguishes the two groups from each other is the more frequent absence of the parents of adolescents who committed sexual offences against children and had a history of sexual abuse. The consequences to be drawn from these results with regard to aggressive sexual delinquency in adolescence are discussed and suggestions are made with regard to further avenues of investigation. PMID- 10837110 TI - A preliminary typology of young sex offenders. AB - Data concerning all young (15-20 years, n = 56) sex offenders (YSOs) subjected to forensic psychiatric investigation in Sweden during 1988-1995 were used in an attempt to construct and validate an introductory YSO typology based solely on offence characteristics. A 5-cluster solution received optimal support from cluster analysis of 15 offence-related variables. A few historical and clinical characteristics varied across clusters. Survival analyses revealed that the clusters differed with respect to sexual but not to violent or general reconviction rates. PMID- 10837111 TI - Evaluation of a teaching package designed to improve teachers' recognition of depressed pupils-a pilot study. AB - This pilot study evaluated the impact of a teaching package for school staff on their ability to recognize depressed pupils within a year group in their school. Sixteen teachers were tested before and after training on their ability to identify depressed pupils in that year. The pupils were screened using a 2-stage procedure to determine caseness for Major Depressive Disorder (MDD). After training, teachers felt more confidant about their ability to recognize depressed pupils but this was not able to be statistically verified because of small teachers numbers. This suggests a promising way forward in recognizing depression in the community. PMID- 10837112 TI - Adolescent egocentrism: a comparison among adolescents and adults. AB - Reformulation of adolescent egocentrism suggests that personal fable and imaginary audience ideations extend into adulthood. To test this proposition, adolescents (aged 14-18) and adults (aged 20-89) completed subscales of the adolescent egocentrism, self-consciousness and interpersonal reactivity scales. An across scale comparison first ensured that adolescent egocentrism measures were comparable across age. Next, MANOVAs revealed higher egocentrism scores for adolescents vs. adults (p<0.05), but no difference between adolescents and younger (19-30) adults after splitting adults into younger (19-30), middle (31 59) and older (60+) subgroups. Results suggest that personal fable and imaginary audience are not confined to adolescence. PMID- 10837113 TI - Family conflict predicts blood pressure changes in African-American adolescents: a preliminary examination. AB - Reviews of the youth literature suggest that the precursors of elevated blood pressure begin in childhood. As such, it may be possible to identify processes among African-American adolescents that are associated with blood pressure changes. This study examined the relationship between family conflict and mean arterial pressure changes in a sample of 39 African-American adolescents. Resting mean arterial pressure, systolic blood pressure, and diastolic blood pressure were assessed at Time 1 and Time 2, separated by a period of 6 months. Assessments of perceived family conflict and negative life events were taken at Time 1. Findings from the multiple regression analyses indicated that family conflict predicted mean arterial blood pressure changes, independent of the effects of age, gender, and body mass index. Although preliminary, these findings highlight the importance of exploring environmental processes that may influence physiological outcomes in adolescents. PMID- 10837114 TI - Eating disorders in patients with cystic fibrosis. AB - This study was designed to examine rates of eating disorders and psychopathology in patients with cystic fibrosis (CF). Fifty-eight CF patients and 43 healthy control participants were evaluated using structured psychiatric interviews and rating scales. Two control participants and no CF patients were diagnosed with an eating disorder. Additionally, 11 CF patients were diagnosed with one or more psychiatric disorders. Group means on the rating scales did not show clinically meaningful elevations in either group. These data indicate no evidence for elevated rates of eating disorders in CF patients. Similarly, rates of other psychiatric disorders in the CF group were not greater than the prevalence reported in the general population. PMID- 10837115 TI - Symbolic representation across domains in preschool children. AB - The study examines children's understanding of notational representation. Children 3 to 5 years old were shown a card containing a notation and told what it said. The children were asked to say what was printed on the card 3 times: when it was under a display named by the notation, when it was under a different display, and finally when it had been returned to the original position. In Study 1, the card contained a word or a picture indicating object identity or a numeral or an analogue indicating quantity. All the children could solve the problems containing pictures, numerals, or analogues, but the word condition was difficult and children believed that the word changed when the card moved to the new display. In Study 2, the comparison between object names and quantities was made more equitable by introducing easy and difficult versions of each condition. This time, there was little difference between cards indicating names and those indicating quantities but large differences in children's ability to solve the problem as a function of their familiarity with the notation that was written. The results point to weaknesses in children's understanding of how representations refer to meanings. PMID- 10837116 TI - Vowel categorization skill and its relationship to early literacy skills among first-grade Quebec-French children. AB - Six-year-old children's ability to categorize words on the basis of vowel categories was examined at the beginning of first grade and again after 6 months of formal schooling. The potential effects of relative proximity of vowels in the vowel space, of syllable structure, and of input phonology were assessed. Also, the effect of literacy instruction on vowel categorization and the relationship of vowel categorization with vowel spelling and reading skill were investigated. Results indicate that the ability to categorize vowels does not develop uniformly but is affected by the degree of spectral/articulatory proximity between vowels, by syllable structure, and potentially by characteristics of the input phonology. Error analyses further indicate that children have fuzzy category boundaries between vowels adjacent on the height continuum. The pattern of results on oral categorization and written tasks suggests a reciprocal relationship. Categorization ability improved after 6 months of schooling. However, vowels that children found more difficult to categorize were also more difficult to read and spell. PMID- 10837117 TI - Children's serial recall errors: implications for theories of short-term memory development. AB - Three experiments examined developmental changes in serial recall of lists of 6 letters, with errors classified as movements, omissions, intrusions, or repetitions. In Experiments 1 and 2, developmental differences between groups of children aged from 7 to 11 years and adults were found in the pattern of serial recall errors. The errors of older participants were more likely to be movements than were those of younger participants, who made more intrusions and omissions. The number of repetition errors did not change with age, and this finding is interpreted in terms of a developmentally invariant postoutput response inhibition process. This interpretation was supported by the findings of Experiment 3, which measured levels of response inhibition in 7-, 9-, and 11-year olds by comparing recall of lists with and without repeated items. Response inhibition remained developmentally invariant, although older children showed greater response facilitation (improved correct recall of adjacent repeated items). Group differences in the patterns of other errors are accounted for in terms of developmental changes in levels of output forgetting and changes in the efficiency of temporal encoding processes. PMID- 10837118 TI - Analysis of mouse fertilin in wild-type and fertilin beta(-/-) sperm: evidence for C-terminal modification, alpha/beta dimerization, and lack of essential role of fertilin alpha in sperm-egg fusion. AB - The sperm surface protein fertilin functions in sperm-egg interaction. On guinea pig and bovine sperm, fertilin is a heterodimer of alpha and beta subunits. Both subunits are initially synthesized as precursors and then proteolytically processed by removing N-terminal domains. Since the mouse is currently the main mammalian species in which fertilization is studied, in the present report, we analyzed the structure, processing, and expression of fertilin in mouse. We found that the processing of mouse fertilin beta occurs during epididymal maturation and involves changes in the cytoplasmic tail domain as well as the N-terminal domains. Although we (R. Yuan et al., 1997, J. Cell Biol. 137, 105-112) and others (M. S. Chen et al., 1999, J. Cell Biol. 144, 549-561) have previously reported that mature fertilin beta is 55-57 kDa, here we show that 55 kDa is an unrelated protein in the sperm extract which cross-reacts with an antibody that recognizes precursor, but not mature, fertilin beta. Comparison of Western blots of wild-type and fertilin beta knockout sperm revealed that authentic, mature fertilin beta is 45 kDa. We also obtained direct evidence that mouse fertilin alpha and beta exist as a heterodimer. In addition, we found that in mice lacking the fertilin beta subunit, fertilin alpha is absent from mature sperm. A widely proposed model for sperm-egg fusion suggests that fertilin alpha is the sperm component that promotes membrane fusion by undergoing a conformational change that exposes a virus-like, hydrophobic fusion peptide. Because sperm lacking fertilin alpha and fertilin beta can fuse with eggs at 50% the wild-type rate, this model is called into question. The results suggest instead that other gamete surface molecules act to promote membrane fusion and that fertilin's role in gamete fusion is in sperm-egg plasma membrane adhesion. PMID- 10837119 TI - Targeting of cre to the Foxg1 (BF-1) locus mediates loxP recombination in the telencephalon and other developing head structures. AB - The use of genetics to study the development of the telencephalon and derivatives such as the cerebral cortex has been limited. The telencephalon begins to form midway through gestation, and targeted mutations in genes suspected of playing roles in its development often lead to early phenotypes that preclude analysis of their role at later stages. This problem can be circumvented using a Cre/loxP recombination system. A mouse line was produced in which cre was targeted to the Foxg1 (BF-1) locus, a gene expressed specifically in the telencephalon and discrete head structures. Crosses between Foxg1-Cre mice and three separate loxP reporter mice generated embryos with recombination patterns matching that expected from the normal pattern of Foxg1 expression. Recombination occurs invariably in the telencephalon, anterior optic vesicle, otic vesicle, facial and head ectoderm, olfactory epithelium, mid-hindbrain junction, and pharyngeal pouches. Recombination in some animals also occurs less efficiently in tissues not known to express Foxg1. We show that the genetic background of the parental mice and the loxP target allele can each contribute to differences in the exact pattern of recombination. Collectively, these data show that Foxg1-Cre mice should be useful in the deletion or ectopic expression of any floxed target gene in a Foxg1-like pattern. PMID- 10837120 TI - Engrailed-1 misexpression in chick embryos prevents apical ridge formation but preserves segregation of dorsal and ventral ectodermal compartments. AB - Using lineage tracers, we recently showed dorsal and ventral ectodermal compartments along the sides of the body in chick embryos. The compartments are formed both in presumptive limb-forming regions where they position the apical ridge and also in presumptive interlimb (flank). Here we show, using a novel technique combining fate mapping and in situ hybridisation, that the ventral compartment coincides with the Engrailed-1 (En-1) domain of expression. This coincidence suggests that En-1 could maintain the ventral compartment and be necessary for apical ridge formation. To test this hypothesis, we ectopically expressed En-1 via retroviral transfer and then examined limb development and cell lineage restriction in the ectoderm. En-1 misexpression can completely prevent formation of both normal limbs and ectopic limbs induced in the flank by application of FGF-2. In both cases, there are no morphological signs of apical ectodermal ridge formation and expression of ridge-associated genes is undetectable. In striking contrast, the lineage restriction between dorsal and ventral ectoderm is not altered. Therefore, En-1 is involved in the regulation of ridge formation but not compartment maintenance. PMID- 10837121 TI - Role of N-myc in the developing mouse kidney. AB - N-myc is a transcription factor expressed in the developing metanephric kidney and other organs. In mice, complete disruption of the N-myc gene results in fetal death on the first day of renal organogenesis. In addition to the null N-myc allele, others have generated a hypomorphic N-myc allele. In this study, combinations of these mutant genes were used to demonstrate that reduction in N myc protein levels correlate with fewer developing glomeruli and collecting ducts in embryonic kidney explants. Histological sections revealed that the mutant kidneys were hypoplastic with normal developing structures. The data indicate that the hypoplasia is due to a reduction in proliferation rather than an increase in apoptosis. Thus, N-myc loss causes a decrease in numbers of ureteric bud tips and developing glomeruli in explants and hypoplastic kidneys in vivo, in a dose-dependent manner. PMID- 10837122 TI - Biphasic elevation of [Ca(2+)](i) in individual human spermatozoa exposed to progesterone. AB - Fluorimetric studies on progesterone-induced [Ca(2+)](i) signalling in mammalian spermatozoa show both the well-characterised [Ca(2+)](i) transient and a subsequent sustained phase. However, the sustained phase is thought to reflect release of the fluorochrome during the acrosome reaction and has not been subject to critical investigation. We have used single-cell imaging of [Ca(2+)](i) to analyse the progesterone-induced [Ca(2+)](i) response in large numbers (>2000) of capacitated, human spermatozoa. In 70% of cells, treatment with progesterone induced a transient increase, which typically peaked within 1 min and decayed with a similar time course. Upon rapid application of progesterone this response peaked within 5-20 s. In 35% of progesterone-treated spermatozoa a sustained elevation of [Ca(2+)](i) occurred, which became discernible during the falling phase of the transient response and persisted for at least 20 min. Both [Ca(2+)](i) responses were localised to the postacrosomal region. Averaging of large numbers of single cell responses generated traces similar to those seen in fluorimetric studies. Although the sustained response was strongly associated with the initial, transient response, a few spermatozoa generated sustained responses that were not preceded by a significant transient response (5% of cells). It is concluded that a genuine biphasic [Ca(2+)](i) signal is activated by progesterone and that the sustained response is a discrete signalling event with biological significance. PMID- 10837123 TI - Negative autoregulation of Mash1 expression in CNS development. AB - Mash1, a neural-specific bHLH transcription factor, is essential for the formation of multiple CNS and PNS neural lineages. Transcription from the Mash1 locus is elevated in mice null for Mash1, suggesting that MASH1 normally acts to repress its own transcription. This activity is contrary to the positive autoregulation of other proneural bHLH proteins. To investigate the mechanisms involved in this process, sequences flanking the Mash1 gene were tested for the ability to mediate negative autoregulation. A Mash1/lacZ transgene containing 36 kb of cis-regulatory sequence exhibits an increase in lacZ expression in the Mash1 mutant background, which phenocopies the observation of transcriptional autoregulation at the endogenous Mash1 locus. Using Mash1/lacZ lines with progressively less cis-acting sequence, autoregulatory responsive elements were demonstrated to colocalize with a previously characterized 1.2-kb CNS enhancer. Mutations of E-box sites within this enhancer did not result in an apparent loss of autoregulation, suggesting that MASH1 does not directly repress its own transcription. Interestingly, these mutations did not indicate any underlying positive auto- or cross-regulation of Mash1. Furthermore, the loss of autoregulation in the Mash1 mutant background is reminiscent of a loss of lateral inhibitory signaling. However, mutations in HES consensus sites, the likely purveyors of Notch-mediated lateral inhibition, do not support a role for these sites in negative autoregulation. We hypothesize that MASH1 normally inhibits its own expression indirectly, possibly through a HES-mediated repression of positive regulators or through novel HES binding sites. PMID- 10837124 TI - Roles for Fgf signaling during zebrafish fin regeneration. AB - Following amputation of a urodele limb or teleost fin, the formation of a blastema is a crucial step in facilitating subsequent regeneration. Using the zebrafish caudal fin regeneration model, we have examined the hypothesis that fibroblast growth factors (Fgfs) initiate blastema formation from fin mesenchyme. We find that fibroblast growth factor receptor 1 (fgfr1) is expressed in mesenchymal cells underlying the wound epidermis during blastema formation and in distal blastemal tissue during regenerative outgrowth. fgfr1 transcripts colocalize with those of msxb and msxc, putative markers for undifferentiated, proliferating cells. A zebrafish Fgf member, designated wfgf, is expressed in the regeneration epidermis during outgrowth. Furthermore, we show that a specific inhibitor of Fgfr1 applied immediately following fin amputation blocks blastema formation, without obvious effects on wound healing. This inhibitor blocks the proliferation of blastemal cells and the onset of msx gene transcription. Inhibition of Fgf signaling during ongoing fin regeneration prevents further outgrowth while downregulating the established expression of blastemal msx genes and epidermal sonic hedgehog. Our findings indicate that zebrafish fin blastema formation and regenerative outgrowth require Fgf signaling. PMID- 10837125 TI - Embryonic muscle development of Convoluta pulchra (Turbellaria-acoelomorpha, platyhelminthes). AB - We studied the embryonic development of body-wall musculature in the acoel turbellarian Convoluta pulchra by fluorescence microscopy using phalloidin-bound stains for F-actin. During stage 1, which we define as development prior to 50% of the time between egg-laying and hatching, actin was visible only in zonulae adhaerentes of epidermal cells. Subsequent development of muscle occurred in two distinct phases: first, formation of an orthogonal grid of early muscles and, second, differentiation of other myoblasts upon this grid. The first elements of the primary orthogonal muscle grid appeared as short, isolated, circular muscle fibers (stage 2; 50% developmental time), which eventually elongated to completely encircle the embryo (stage 3; at 60% of total developmental time). The first primary longitudinal fibers appeared later, along with some new primary circular fibers, by 60-63% of total developmental time (stage 4). From 65 to 100% of total developmental time (stages 5 to 7), secondary fibers, using primary fibers as templates, arose; the number of circular and longitudinal muscles thus increased, and at the same time parenchymal muscles began appearing. Hatchlings (stage 8) possessed about 25 circular and 30 longitudinal muscles as well as strong parenchymal muscles. The remarkable feature of the body wall of many adult acoel flatworms is that longitudinal muscles bend medially and cross each other behind the level of the mouth. We found that this development starts shortly after the appearance of the ventral mouth opening within the body wall muscle grid. The adult organization of the body-wall musculature consists of a grid of several hundred longitudinal and circular fibers and a few diagonal muscles. Musculature of the reproductive organs developed after hatching. Thus, extensive myogenesis must occur also during postembryonic development. Comparison between the turbellarians and the annelids suggests that formation of a primary orthogonal muscle grid and its subsequent use as a template for myoblast differentiation are the two basic developmental phases in vermiform Spiralia if not in the Bilateria as a whole. Finally, our new data suggest that for the Acoela the orthogonal primary patterning of longitudinal and circular muscles in the body wall is achieved without using originally positional information of the nervous system. PMID- 10837126 TI - The bHLH class protein pMesogenin1 can specify paraxial mesoderm phenotypes. AB - A new bHLH gene from mouse that we call pMesogenin1 (referring to paraxial mesoderm-specific expression and regulatory capacities) and its candidate ortholog from Xenopus were isolated and studied comparatively. In both organisms the gene is specifically expressed in unsegmented paraxial mesoderm and its immediate progenitors. A striking feature of pMesogenin1 expression is that it terminates abruptly in presumptive somites (somitomeres). Somitomeres rostral to the pMesogenin1 domain strongly upregulate expression of pMesogenin's closest known paralogs, MesP1 and MesP2 (Thylacine1/2 in Xenopus). Subsequently, the most rostral somitomere becomes a new somite and expression of MesP1/2 is sharply downregulated before this transition. Thus, expression patterns of these bHLH genes, together with that of an additional bHLH gene in the mouse, Paraxis, collectively define discrete but highly dynamic prepatterned subdomains of the paraxial mesoderm. In functional assays, we show that pMesogenin1 from either mouse or frog can efficiently drive nonmesodermal cells to assume a phenotype with molecular and cellular characteristics of early paraxial mesoderm. Among genes induced by added pMesogenin1 is Xwnt-8, a signaling factor that induces a similar repertoire of marker genes and a similar cellular phenotype. Additional target genes induced by pMesogenin1 are ESR4/5, regulators known to play a significant role in segmentation of paraxial mesoderm (W. C. Jen et al., 1999, Genes Dev. 13, 1486-1499). pMesogenin1 differs from other known mesoderm-inducing transcription factors because it does not also activate a dorsal (future axial) mesoderm phenotype, suggesting that pMesogenin1 is involved in specifying paraxial mesoderm. In the context of the intact frog embryo, ectopic pMesogenin1 also actively suppressed axial mesoderm markers and disrupted normal formation of notochord. In addition, we found evidence for cross-regulatory interactions between pMesogenin1 and T-box transcription factors, a family of genes normally expressed in a broader pattern and known to induce multiple types of mesoderm. Based on our results and results from prior studies of related bHLH genes, we propose that pMesogenin1 and its closest known relatives, MesP1/2 (in mouse) and Thylacine1/2 (in Xenopus), comprise a bHLH subfamily devoted to formation and segmentation of paraxial mesoderm. PMID- 10837127 TI - The maternal effect mutation sesame affects the formation of the male pronucleus in Drosophila melanogaster. AB - After entering the oocyte and before the formation of the diploid zygote, the sperm nucleus is transformed into a male pronucleus, a process that involves a series of conserved steps in sexually reproducing animals. Notably, a major modification of the male gamete lies in the decondensation of the highly compact sperm chromatin. We present here the phenotype of sesame (ssm), a maternal effect mutation which affects the formation of the male pronucleus in Drosophila melanogaster. Homozygous ssm(185b) females produce haploid embryos which develop with only the maternally derived chromosomes. These haploid embryos die at the end of embryogenesis. Cytological analyses of the fertilization in eggs laid by ssm(185b) mutant females showed that both pronuclear migration and pronuclear apposition occurred normally. However, a dramatic alteration of the male pronucleus by which its chromatin failed to fully decondense was systematically observed. Consequently, the affected male pronucleus does not enter the first mitotic spindle, which is organized around only the maternally derived chromosomes. Immunodetection of lamina antigens indicates that a male pronuclear envelope is able to form around the partially decondensed paternal chromatin. This suggests that the maternally provided sesame(+) function is required for a late stage of sperm chromatin remodeling. PMID- 10837128 TI - Xbra3 induces mesoderm and neural tissue in Xenopus laevis. AB - Homologues of the murine Brachyury gene have been shown to be involved in mesoderm formation in several vertebrate species. In frogs, the Xenopus Brachyury homologue, Xbra, is required for normal formation of posterior mesoderm. We report the characterisation of a second Brachyury homologue from Xenopus, Xbra3, which has levels of identity with mouse Brachyury similar to those of Xbra. Xbra3 encodes a nuclear protein expressed in mesoderm in a temporal and spatial manner distinct from that observed for Xbra. Xbra3 expression is induced by mesoderm inducing factors and overexpression of Xbra3 can induce mesoderm formation in animal caps. In contrast to Xbra, Xbra3 is also able to cause the formation of neural tissue in animal caps. Xbra3 overexpression induces both geminin and Xngnr 1, suggesting that Xbra3 can play a role in the earliest stages of neural induction. Xbra3 induces posterior nervous tissue by an FGF-dependent pathway; a complete switch to anterior neural tissue can be effected by the inhibition of FGF signalling. Neither noggin, chordin, follistatin, nor Xnr3 is induced by Xbra3 to an extent different from their induction by Xbra nor is BMP4 expression differentially affected. PMID- 10837130 TI - Distinct mechanisms triggering glial differentiation in Drosophila thoracic and abdominal neuroblasts 6-4. AB - Neurons and glia are produced in stereotyped patterns after neuroblast cell division during development of the Drosophila central nervous system. The first cell division of thoracic neuroblast 6-4 (NB6-4T) is asymmetric, giving rise to a glial precursor cell and a neuronal precursor cell. In contrast, abdominal NB6-4 (NB6-4A) divides symmetrically to produce two glial cells. To understand the relationship between cell division and glia-neuron cell fate determination, we examined and compared the effects of known cell division mutations on the NB6-4T and NB6-4A lineages. Based on observation of expression of glial fate determination and early glial differentiation genes, the onset of glial differentiation occurred in NB6-4A but not in NB6-4T when both cell cycle progression and cytokinesis were genetically arrested. On the other hand, glial differentiation started in both lineages when cytokinesis was blocked with intact cell cycle progression. These results showed that NB6-4T, but not NB6-4A, requires cell cycle progression for acquisition of glial fate, suggesting that distinct mechanisms trigger glial differentiation in the different lineages. PMID- 10837129 TI - MAP kinase, a universal suppressor of sperm centrosomes during meiosis? AB - We reported previously that inhibition of MAP kinase during meiosis in Urechis caupo eggs caused premature sperm aster formation and we reviewed indirect evidence that the suppression of sperm asters by MAPK during meiosis might be a universal mechanism (M. C. Gould and J. L. Stephano, 1999, Dev. Biol. 216, 348 358). We tested this proposition with oyster (Crassostrea gigas) and starfish (Asterina miniata) eggs, utilizing the MEK inhibitors U0126 and PD98059. Centrosomes, asters, and meiotic spindles were visualized by normal epifluorescence and confocal microscopy following indirect immunocytochemical staining for anti-beta-tubulin. When MAPK activation was inhibited, sperm asters in both species developed prematurely and tended to move toward the egg centrosomes, sometimes even fusing with the egg spindle or centrosomes. Meiotic spindles and polar body formation were also abnormal when MAPK was inhibited. PMID- 10837131 TI - Cortical actin filaments form rapidly during photopolarization and are required for the development of calcium gradients in Pelvetia compressa zygotes. AB - Previous research has shown that cortical gradients of cytosolic Ca(2+) are formed during the photopolarization of Pelvetia compressa zygotes, with elevated Ca(2+) on the shaded hemisphere that will become the site of rhizoid germination. We report here that the marine sponge toxin, latrunculin B, which blocks photopolarization at nanomolar concentrations, inhibited the formation of the light-driven Ca(2+) gradients. Using low concentrations of microinjected fluorescent phalloidin as a tracer for actin filaments, we found that exposure to light induced a striking increase in actin filaments in the cells as indicated by an increase in fluorescence. The increase was quantified in the cortex, where it was most apparent, and the fluorescence there was found to increase by about a factor of 3. This increase in cortical phalloidin fluorescence was inhibited by latrunculin B at the same concentration required to inhibit Ca(2+) gradient formation and photopolarization. The distribution of the increasing phalloidin fluorescence was uniform with respect to the developing rhizoid-thallus axis during the formation of the axis, and no intense patches of fluorescence were observed. After germination, fluorescence suggestive of an apical ring of actin filaments was seen near the rhizoid tip. Finally, inhibitor studies indicated that myosin may be involved in the photopolarization process. PMID- 10837132 TI - Imaginal pioneers prefigure the formation of adult thoracic muscles in Drosophila melanogaster. AB - In insects, specialized mesodermal cells serve as templates to organize myoblasts into distinct muscle fibers during embryogenesis. In the grasshopper embryo, large mesodermal cells called muscle pioneers extend between the epidermal attachment points of future muscle fibers and serve as foci for myoblast fusion. In the Drosophila embryo, muscle founder cells serve a similar function, organizing large numbers of myoblasts into larval muscles. During the metamorphosis of Drosophila, nearly all larval muscles degenerate and are replaced by a set of de novo adult muscles. The extent to which specialized mesodermal cells homologous to the founders and pioneers of the insect embryo are involved in the development of adult-specific muscles has yet to be established. In the larval thorax, the majority of imaginal myoblasts are associated with the imaginal discs. We report here the identification of a morphologically distinct class of disc-associated myoblasts, which we call imaginal pioneers, that prefigures the formation of at least three adult-specific muscles, the tergal depressor of the trochanter and dorsoventral muscles I and II. Like the muscle pioneers of the grasshopper, the imaginal pioneers attach to the epidermis at sites where the future muscle insertions will arise and erect a scaffold for developing adult muscles. These findings suggest that a prior segregation of imaginal myoblasts into at least two populations, one of which may act as pioneers or founders, must occur during development. PMID- 10837133 TI - Retinoic acid in the formation of the dorsoventral retina and its central projections. AB - Dynamic expression patterns of four retinoid-metabolizing enzymes create rapidly changing retinoic acid (RA) patterns in the emerging eye anlage of the mouse. First, a RA-rich ventral zone is set up, then a RA-poor dorsal zone, and finally a tripartite organization consisting of dorsal and ventral RA-rich zones separated by a horizontal RA-poor stripe. This subdivision of the retina into three RA concentration zones is directly visible as beta-galactosidase labeling patterns in retinas of RA-reporter mice. Because the axons of retinal ganglion cells transport the reporter product anterogradely, the central projections from dorsal and ventral retina can be visualized as two heavily labeled axon bundles. Comparisons of the axonal labeling with physiologic recordings of visual topography in the adult mouse show that the labeled axons represent the upper and the lower visual fields. The RA-poor stripe develops into a broad horizontal zone of higher visual acuity. Comparisons of the retina labeling with eye-muscle insertions show that the axis of the RA pattern lines up with the dorsoventral axis of the oculomotor system. These observations indicate that the dorsoventral axis of the embryonic eye anlage determines the functional coordinates of both vision and eye movements in the adult. PMID- 10837134 TI - Metamorphosis of tangential visual system neurons in Drosophila. AB - To learn about construction of the adult nervous system, we studied the differentiation of imaginal neurons in the Drosophila visual system. OL2-A and OL3 are tangential neurons that display dFMRFa neuropeptide gene expression in adults but not in larvae. The two large OL2-A neurons are generated near the end of the embryonic period and already show morphological differentiation at the start of metamorphosis. The numerous small OL3 neurons are generated postembryonically and first detected later in metamorphosis. The onset of dFMRFa transcription coincides with that of neuropeptide accumulation in OL2-A neurons, but it precedes peptide accumulation in the OL3 neurons by days. Altering each of the five conserved sequences within the minimal 256-bp OL dFMRFa enhancer affected in vivo OL transcriptional activity in two cases: alteration of a TAAT element greatly diminished and alteration of a 9-bp tandem repeat completely abolished OL2-A/OL3 reporter activity. A 46-bp concatamer containing the TAAT element, tested separately, was not active in OL neurons. We propose a model of neuronal differentiation at metamorphosis that features developmental differences between classes of imaginal neurons. PMID- 10837135 TI - Differential involvement of Na(+),K(+)-ATPase isozymes in preimplantation development of the mouse. AB - Na(+),K(+)-ATPase plays an essential role in mammalian blastocoel formation (cavitation) by driving trans-epithelial sodium transport. Previously, the alpha1 and beta1 subunit isoforms of this enzyme were identified in preimplantation mouse embryos and were assumed to be responsible for this function. Here we show that mRNAs encoding an additional alpha subunit isoform (alpha3) and the remaining two beta subunit isoforms are also present in preimplantation embryos. Whereas alpha3 mRNA accumulates between the four-cell and the blastocyst stages and thus results from embryonic transcription, the same could not be demonstrated for beta2 and beta3 mRNAs. Immunoblot analyses confirmed that these subunits are present in cavitating embryos. Using confocal immunofluorescence microscopy we found that alpha1 and beta1 subunits are concentrated in the basolateral membranes of the trophectoderm while being equally distributed in plasma membranes of the inner cell mass. In contrast, alpha3, beta2, and beta3 subunits were not detected in plasma membranes. Our current assessment, therefore, is that as many as six isozymes of Na(+),K(+)-ATPase could be involved in preimplantation development although it is primarily the alpha1beta1 isozyme that is responsible for blastocoel formation. Our findings imply that the regulation of sodium transport within the preimplantation mouse embryo is more complex than had been appreciated. PMID- 10837136 TI - The Ki-67 protein: fascinating forms and an unknown function. AB - The Ki-67 protein is a nuclear and nucleolar protein, which is tightly associated with somatic cell proliferation. Antibodies raised against the human Ki-67 protein paved the way for the immunohistological assessment of cell proliferation, particularly useful in numerous studies on the prognostic value of cell growth in clinical samples of human neoplasms. The primary structure revealed potential phosphorylation site for a range of essential kinases, PEST sequences, and a forkhead-associated domain, which are features present in a variety of cell-cycle-regulating proteins, but information about the position of the Ki-67 protein within the protein network that drives the cell cycle remained scarce. There is now evidence that posttranslational modifications based on phosphorylation by cdc2 kinase and PKC accompany the remarkable redistribution of the Ki-67 protein from the interior of the nucleus to the perichromosomal layer during mitosis and vice versa. The discovery of Ki-67 equivalents in other species is advantageous for a precise and cross-species investigation of the structural requirements for its yet unknown function. The recently published data add new pieces to the challenging puzzle of this multifaceted protein, which are waiting to be put together. PMID- 10837137 TI - Uncoupling of gate and fence functions of MDCK cells by the actin-depolymerizing reagent mycalolide B. AB - The tight junction serves as a paracellular gate to seal the paracellular space of apposing cells and as a molecular fence to prevent diffusion of membrane proteins and lipids in epithelial cells. Although involvement of the actin cytoskeleton has been considered to be important in these two functions, it remains to be elucidated whether both functions are regulated in a coupled manner or differentially by actin. Treatment of highly polarized MDCK cells with mycalolide B (MB), a recently developed actin-depolymerizing reagent, induced a decrease of transepithelial resistance in a dose- and time-dependent manner with reversibility when the reagent was washed out. Changes in cytoskeletal actin, such as a reduction of cortical actin, irregularity of stress fibers, and punctated actin aggregates, were observed after MB treatment. However, the fence function, as studied by diffusion of apically labeled sphingomyelin/BSA complex, remained intact in the MB-treated MDCK cells. Localization of junctional molecules and apical marker proteins such as E-cadherin, ZO-1, and 114-kDa protein was shown to be unaffected. Furthermore, freeze-fracture study showed apparent tight junction strands. Collectively, MB treatment abolished the paracellular gate but not the fence function of MDCK cells, suggesting that cytoskeletal actin may play differential roles in the gate and fence functions of the tight junction. PMID- 10837138 TI - Platelet-derived growth factor-mediated signaling through the Shb adaptor protein: effects on cytoskeletal organization. AB - The Src homology (SH) 2 domain adaptor protein Shb has previously been shown to interact with the platelet-derived growth factor (PDGF)-beta receptor. In this study we show an association between Shb and the PDGF-alpha receptor which is mediated by the SH2 domain of Shb and involves tyrosine residue 720 in the kinase insert domain of the receptor. To assess the role of Shb in PDGF-mediated signaling, we have overexpressed wild-type Shb or Shb carrying a mutation (R522K) which renders the SH2 domain inactive, in Patch mouse (PhB) fibroblasts expressing both PDGF receptors (PhB/Ralpha). Overexpression of wild-type Shb, but not the R522K Shb mutant, affected PDGF-mediated reorganization of the cytoskeleton by decreasing membrane ruffle formation and stimulating the generation of filopodia relative the parental control cells. In addition, the PDGF-induced receptor-associated phosphatidylinositol 3'-kinase activity and phosphorylation of Akt was similar in both PhB/Ralpha/Shb and PhB/Ralpha/ShbR522K cells compared with the parental control, whereas the activation of Rac in response to PDGF-BB was diminished only in the PhB/Ralpha/Shb cells. We conclude that Shb plays a role in PDGF-dependent regulation of certain cytoskeletal changes by modulating the ability of PDGF to activate Rac. PMID- 10837139 TI - The disruption of adherens junctions is associated with a decrease of E-cadherin phosphorylation by protein kinase CK2. AB - The down-regulation of E-cadherin is a common event in carcinogenesis. Phosphorylation/dephosphorylation is one posttranscriptional process which may regulate intercellular junctions. Here we show that in okadaic acid-treated keratinocytes, E-cadherin expression is shifted from the membrane to the cytoplasm, preventing cells from forming aggregates. These changes of E-cadherin localization and function are associated with a decrease in its phosphorylation state. The decrease in E-cadherin phosphorylation was essentially detected in okadaic acid-treated cell lysates isolated from 0.5% Triton-soluble fraction and not in the Triton-insoluble fraction linked to the cytoskeleton, suggesting a role of E-cadherin phosphorylation in cell-cell interactions. E-cadherin was markedly phosphorylated by CK2, either the purified recombinant enzyme or the endogenous enzyme. Using specific CK2 inhibitors such as heparin and 5, 6 dichloro-1-beta-d-ribofuranosylbenzimidazole, endogenous CK2 was confirmed as the main enzyme phosphorylating E-cadherin. The decrease in E-cadherin phosphorylation by endogenous CK2 was not restored by the addition of purified CK2, confirming that it is not due to a defect in CK2 expression or to its reduced activity, but rather to the incapacity of CK2 to phosphorylate E cadherin. The co-immunoprecipitation and colocalization of E-cadherin and CK2 suggests that CK2 may play a critical role in the maintenance of epidermis cohesion. PMID- 10837140 TI - The nucleoside diphosphate kinase activity of DRnm23 is not required for inhibition of differentiation and induction of apoptosis in 32Dcl3 myeloid precursor cells. AB - DRnm23 belongs to a multigene family which includes nm23-H1, the first bona fide metastasis suppressor gene, nm23-H2, nm23-H4, and nm23-H5. Like nm23-H1, nm23-H2, and nm23-H4, DRnm23 possesses nucleoside diphosphate kinase (NDPK) activity. Upon overexpression in myeloid precursor 32Dcl3 cells, DRnm23 inhibits granulocytic differentiation and promotes apoptosis. Two specific mutants of DRnm23 (H134Q and S136P), at residues required for the NDPK activity, inhibit differentiation and promote apoptosis of 32Dcl3 cells. By contrast, substitution of serine 61 with proline (S61P) or deletion of the RGD domain (DeltaRGD) abrogates the effects of wild-type DRnm23. Like wild-type DRnm23, all four mutants show a predominantly mitochondrial subcellular localization. These studies indicate that the enzymatic activity of DRnm23 is not required for the effects observed in 32Dcl3 cells. Moreover, the inability of the S61P and DeltaRGD DRnm23 mutants to inhibit differentiation and promote apoptosis may be due to defective protein-protein interactions at the mitochondria, the predominant site of DRnm23 subcellular localization. PMID- 10837141 TI - The nuclear DEAD box RNA helicase p68 interacts with the nucleolar protein fibrillarin and colocalizes specifically in nascent nucleoli during telophase. AB - The DEAD box protein, p68, is an established RNA-dependent ATPase and RNA helicase in vitro, but neither the physiological function of this protein nor the macromolecules with which it interacts are known. Using a yeast two-hybrid screen, we identified the nucleolar protein, fibrillarin, as a protein that interacts with p68. Coimmunoprecipitation experiments confirmed that p68 and fibrillarin can form complexes in cellular extracts, and deletion analysis identified regions in each protein responsible for mediating the interaction. Immunofluorescence studies using confocal microscopy revealed that, in interphase cells, while fibrillarin is predominantly nucleolar, p68 shows a diffuse granular nuclear staining but is largely excluded from the nucleoli. Strikingly, both proteins colocalize in nascent nucleoli during late telophase. These data are consistent with a role for p68 either in postmitotic nucleolar reassembly or in the activation of ribosomal DNA transcription/preribosomal RNA processing during telophase and suggest that differential subnuclear compartmentalization may be a mechanism by which interaction of p68 with fibrillarin is regulated in the cell. PMID- 10837142 TI - Enhanced calcium transients in glial cells in neonatal cerebellar cultures derived from S100B null mice. AB - S100B is the major low-affinity Ca(2+)-binding protein in astrocytes. In order to study the role of S100B in the maintenance of Ca(2+) homeostasis, we generated S100B null mice by a targeted inactivation of the S100B gene. Absence of S100B expression was demonstrated by Northern and Western blotting for S100B mRNA and protein, respectively, and immunoperoxidase staining of sections of various brain regions. S100B null mice were viable, fertile, and exhibited no overt behavioral abnormalities up to 12 months of age. On the basis of light microscopy and immunohistochemical staining, there were no discernable alterations in the distribution and morphology of astrocytes or neurons in sections of adult brains of these mice. Astrocytes in cerebellar cultures derived from 6-day-old S100B null mice exhibited enhanced Ca(2+) transients in response to treatment with KCl or caffeine. On the other hand, granule neurons, in the same cultures, exhibited normal Ca(2+) transients in response to treatment with KCl, caffeine, or N-methyl d-aspartate. These results demonstrate a specific decrease in Ca(2+)-handling capacity in astrocytes derived from S100B null mice and suggest that S100B plays a role in the maintenance of Ca(2+) homeostasis in astrocytes. PMID- 10837143 TI - During apoptosis of HL-60 and U-937 cells caspases are activated independently of dissipation of mitochondrial electrochemical potential. AB - Collapse of the mitochondrial potential (DeltaPsi(m)) during apoptosis has been linked with a release of cytochrome c and apoptosis-inducing factor (AIF) and activation of caspases. Using a laser scanning cytometer (LSC), an instrument that allows one to measure the same cells twice, first when they are alive and subsequently after their permeabilization, we explored whether dissipation of DeltaPsi(m) (measured supravitally) is a prerequisite for the activation of caspases (detected after cell fixation). Apoptosis of HL-60 cells was induced either by TNF-alpha combined with cycloheximide (CHX) or by the DNA topoisomerase I inhibitor camptothecin (CPT) and of U-937 cells by CPT, and DeltaPsi(m) was measured using the carbocyanine fluorochrome DiIC(1) (5). The marker of caspase activation was specific cleavage of poly(ADP) ribose polymerase (PARP) detected immunocytochemically. After 30 or 60 min treatment with TNF-alpha + CHX or 60 or 120 min with CPT a considerable proportion of cells (20-40%) demonstrated PARP cleavage with no evidence of DeltaPsi(m) collapse. Also present in these cultures (3-20%) were cells with collapsed DeltaPsi(m) whose PARP was not cleaved. The results provide direct evidence that in HL-60 and U-937 cells treated with TNF alpha + CHX or CPT the dissipation of DeltaPsi(m) is not required for activation of caspases and these two events are independent of each other. PMID- 10837144 TI - Laminin alpha1-chain shows a restricted distribution in epithelial basement membranes of fetal and adult human tissues. AB - Two novel monoclonal antibodies were raised and used to study the expression of laminin (Ln) alpha1-chain in developing and adult human tissues. In both fetal and adult kidney, a distinct immunoreactivity was seen in basement membranes (BM) of most proximal tubules but not in the distal tubular or glomerular BM or in the basal laminae of blood vessels. Immunoprecipitation of metabolically labeled cultured human renal proximal tubular cells showed an abundant production and deposition of Ln alpha1-chain to the extracellular matrix, suggestive of an epithelial origin of kidney Ln-1. Quantitative cell adhesion experiments with JAR choriocarcinoma cells showed that purified human Ln-1 is a good substrate for cell adhesion that it is differently recognized by integrin receptors when compared to mouse Ln-1. In fetal and adult testes immunoreactivity was solely confined to BM of the seminiferous epithelium. In the airways BM-confined reaction was only seen in fetal budding bronchial tubules (16-19 weeks) at the pseudoglandular stage of development. In the skin a distinct immunoreactivity was confined to BM of developing hair buds but not in epithelial BMs of adult epidermis or of epidermal appendages. In other adult tissues, immunoreactivity was found in BMs of thyroid, salivary, and mammary glands as well as in BMs of endometrium and endocervix, but not of ectocervix or vagina. No immunoreactivity was found in BMs of most of the digestive tract, including the liver and pancreas, except for BMs of esophageal submucosal glands and duodenal Brunner's glands. In fetal specimens, BMs of the bottoms of the intestinal and gastric glands were positive. Basal laminae of blood vessels were generally negative for Ln alpha1 chain with the exception of specimens of both fetal and adult central nervous system in which immunoreactivity for Ln alpha1 chain was prominently confined to capillary walls. The results suggest that outside the central nervous system, Ln alpha1 chain shows a restricted and developmentally regulated expression in BMs of distinct epithelial tissues. PMID- 10837145 TI - Human G protein gamma(11) and gamma(14) subtypes define a new functional subclass. AB - The mammalian gamma subunit family consists of a minimum of 12 members. Analysis of the amino acid sequence conservation suggests that the gamma subunit family can be divided into three distinct subclasses. The division of the gamma subunit family into these classes is based not only on amino acid homology, but also to some extent on functional similarities. In the present study, two new members of the gamma subunit family, the gamma(11) and gamma(14) subunits, are identified and characterized in terms of their expression and function. The gamma(11) and gamma(14) subunits are most closely related to the gamma(1) subunit and share similar biochemical properties, suggesting their inclusion in class I. However, despite their close phylogenetic relationship and similar biochemical properties, the gamma(1), gamma(11), and gamma(14) subunits exhibit very distinct expression patterns, suggesting that class I should be further subdivided and that the signaling functions of each subgroup are distinct. In this regard, the gamma(11) and gamma(14) subunits represent a new subgroup of farnesylated gamma subunits that are expressed outside the retina and have functions other than phototransduction. PMID- 10837146 TI - Negative regulation of selected bHLH proteins by eHAND. AB - The bHLH protein eHAND plays an important role in the development of extraembryonic, mesodermal, and cardiac cell lineages, presumably through heterodimerization with other HLH proteins and DNA binding. In this study, we have identified a novel transcriptional activity of eHAND. In transient transfection assays, eHAND is a potent inhibitor of activation by some but not all bHLH proteins. eHAND can prevent E-box DNA binding by these bHLH proteins. Interestingly, eHAND can also strongly inhibit transactivation activity by a MyoD approximately E47 tethered dimer, which suggests a distinct mechanism of action. eHAND also inhibits MyoD-dependent skeletal muscle cell differentiation and expression of the muscle-specific myosin heavy chain protein. In addition, we show that eHAND can repress activity of the natural p75LNGFR promoter, whose expression overlaps that of eHAND and dHAND. The inhibitory activity of eHAND may be attributed to multiple mechanisms, such as the ability to act as a corepressor, the presence of a repression domain, and its ability to sequester E proteins in an inactive complex. Based upon its inhibitory effect on bHLH proteins and cellular differentiation, we propose that eHAND may function by several mechanisms to promote placental giant cell proliferation by negatively regulating the activities of the bHLH protein MASH-2. PMID- 10837147 TI - WW- and SH3-domain interactions with Epstein-Barr virus LMP2A. AB - Epstein-Barr virus (EBV) is a human herpesvirus which establishes a lifelong latent infection in B lymphocytes. Latent membrane protein 2A (LMP2A) is expressed in both humans with EBV latent infection and EBV immortalized cell lines grown in culture. Previous studies have shown that the amino terminal domain of LMP2A, which contains eight tyrosines, associates with a variety of cellular proteins via SH2-phosphotyrosine interactions. Also contained within the LMP2A amino terminal domain are five proline-rich regions, three of which possess the PxxP core consensus sequence required for interacting with SH3 domains and two of which possess the PPxY core consensus sequence (PY motif) required for interacting with class I type WW domains. In the current study, the ability of LMP2A to interact with either modular SH3 or WW domains was investigated. The results of these studies indicate that the two LMP2A PY motifs interact strongly with representative class I WW domains, but not with representative class II WW domains. In contrast, no interactions were detected between LMP2A and any of the five different SH3 domains tested. These data demonstrate that a subset of the conserved proline-rich motifs within the amino terminus of LMP2A can potentially mediate interactions with cellular proteins and may play a role in EBV-mediated latency and/or transformation. PMID- 10837148 TI - Injections of porcine sperm extracts trigger fertilization-like calcium oscillations in oocytes of a marine worm. AB - The precise mechanisms by which sperm trigger calcium transients in eggs or oocytes during fertilization remain unknown. Based on time-lapse confocal microscopy, we show that intracellular injections of porcine sperm extracts cause the oocytes of a marine nemertean worm to undergo repetitive calcium oscillations resembling those obtained during normal fertilizations. Such findings are consistent with the view that fertilization involves a soluble sperm factor (SF) which is capable of eliciting calcium transients without binding to externally situated receptors on the oocyte plasmalemma. This study also describes for the first time the wave-like propagation patterns of SF-induced calcium transients that are generated in a heterologous combination of gametes obtained from different phyla of animals. Such cross-reactivity between distantly related taxa suggests that the intracellular signaling pathways triggered by sperm factors can be well conserved. PMID- 10837149 TI - Phylogeny and biogeography of serolid isopods (Crustacea, Isopoda, Serolidae) and the use of ribosomal expansion segments in molecular systematics. AB - In this study, a molecular phylogenetic hypothesis for 16 species of serolid isopods (Crustacea, Isopoda, Serolidae) from Antarctic waters, the deep sea, South America, and Australia is presented. The genes used are a 500-bp fragment of the mitochondrial LSU rRNA gene and a 700-bp fragment located in the variable region V4 of the nuclear SSU rRNA gene. The species composition and monophyly of morphologically defined genera of which several members were available are confirmed by the molecular data (Ceratoserolis, Spinoserolis, and Cuspidoserolis). The molecular data also support the redefinition of Frontoserolis s.l. and Serolella and the erection of the new genera Septemserolis and Paraserolis, as proposed by W]agele. The relationship among several genera is resolved differently in the molecular hypothesis than in the two existing morphological hypotheses, however. The molecular phylogeny may have important consequences for understanding the biogeography of the Serolidae, indicating that all Antarctic species in this study form a monophyletic group which has probably derived from species with closest extant relatives in South America. All 3 species included in this study living today in deep waters (>2000 m) of the Southern Ocean are most closely related to species living on the Antarctic shelf, so that parallel colonization of the deep sea by way of polar submergence can be reconstructed. In this study, a V4 expansion segment is reported which exceeds the longest crustacean sequences known until now by more than 270 bp. Although the V4 expansion segment has proven useful for phylogenetic purposes in this study, there is circumstantial evidence that its mechanism of evolution may depend not only on inheritance of single-site substitutions, making its routine use in phylogenetic studies potentially dangerous. PMID- 10837150 TI - Molecular phylogeny, taxonomy, and evolution of nonmarine lineages within the American grapsoid crabs (Crustacea: brachyura). AB - Grapsoid crabs are best known from the marine intertidal and supratidal. However, some species also inhabit shallow subtidal and freshwater habitats. In the tropics and subtropics, their distribution even includes mountain streams and tree tops. At present, the Grapsoidea consists of the families Grapsidae, Gecarcinidae, and Mictyridae, the first being subdivided into four subfamilies (Grapsinae, Plagusiinae, Sesarminae, and Varuninae). To help resolve phylogenetic relationships among these highly adaptive crabs, portions of the mitochondrial genome corresponding to the 16S rRNA gene were sequenced for all grapsoid genera occurring in America. The resulting phylogeny confirms most of the present grapsid subfamilies but suggests reclassification of some of the genera and recognition of new taxonomic units. The two American gecarcinid genera might not represent a sister group to the Grapsidae but rather appear to have evolved within the latter. Colonization of inland habitats evolved in several lineages of the grapsoids, resulting in various forms of nonmarine life and different degrees of independence from the sea. PMID- 10837151 TI - Phylogeny and historical biogeography of the loliginid squids (Mollusca: cephalopoda) based on mitochondrial DNA sequence data. AB - The cephalopod taxon Loliginidae (Cephalopoda: Myopsida) is a species-rich group of tropical and temperate shallow-water squids, many of which are commercial fisheries objects and neurophysiological research organisms. The worldwide distribution of these squids could make Loliginidae a useful case study in shallow-water marine biogeography, but the phylogeny of the group is unknown. To clarify loliginid phylogeny, regions of two mitochondrial genes (the 16S rRNA and the cytochrome c oxidase subunit I genes) were sequenced for members of 19 loliginid species and several outgroups. Maximum-parsimony and maximum-likelihood analyses were performed on a combined data set, as well as on each data set individually. Analyses of the combined data support loliginid monophyly and reveal four clades-one consisting primarily of species in American waters from two genera, one composed of 3 east Atlantic species, one consisting of the bioluminescent loliginids (Uroteuthis sensu Vecchione et al., 1998) plus Loliolus japonica, and one represented by a Loligo (Alloteuthis) subulata-Lolliguncula mercatoris pair. The likelihood of the unconstrained maximum-likelihood tree is not significantly better than the likelihoods of the best trees constrained to Sepioteuthis monophyly or Uroteuthis monophyly, but there is significant support for Lolliguncula polyphyly. Tests of alternative hypotheses of loliginid cladogenesis suggest that cladogenesis within Loliginidae is correlated with the widening of the Atlantic and the closure of the Tethys Sea, although dispersal from the Indo-West Pacific is a reasonable explanation for the origin of the clade of American loliginines. PMID- 10837152 TI - The impact of parsimony weighting schemes on inferred relationships among toucans and neotropical barbets (Aves: piciformes). AB - The development of new schemes for weighting DNA sequence data for phylogenetic analysis continues to outpace the development of consensus on the most appropriate weights. The present study is an exploration of the similarities and differences between results from 22 character weighting schemes when applied to a study of barbet and toucan (traditional avian families Capitonidae and Ramphastidae) phylogenetic relationships. The dataset comprises cytochrome b sequences for representatives of all toucan and Neotropical barbet genera, as well as for several genera of Paleotropical barbets. The 22 weighting schemes produced conflicting patterns of relationship among taxa, often with conflicting patterns each receiving strong bootstrap support. Use of multiple weighting schemes helped to identify the source within the dataset (codon position, transitions, transversions) of the various putative phylogenetic signals. Importantly, some phylogenetic hypotheses were consistently supported despite the wide range of weights employed. The use of phylogenetic frameworks to summarize the results of these multiple analyses proved very informative. Relationships among barbets and toucans inferred from these data support the paraphyly of the traditional Capitonidae. Additionally, these data support paraphyly of Neotropical barbets, but rather than indicating a relationship between Semnornis and toucans, as previously suggested by morphological data, most analyses indicate a basal position of Semnornis within the Neotropical radiation. The cytochrome b data also allow inference of relationships among toucans. Supported hypotheses include Ramphastos as the sister to all other toucans, a close relationship of Baillonius and Pteroglossus with these two genera as the sister group to an (Andigena, Selenidera) clade, and the latter four genera as a sister group to Aulacorhynchus. PMID- 10837153 TI - Phylogenetic analysis of the genus Cheilosia (Diptera, Syrphidae) using mitochondrial COI sequence data. AB - The genus Cheilosia is one of the most diverse and speciose genera of Syrphidae (Diptera). The phylogenetic relationships of the hoverfly genus Cheilosia was investigated for the first time using molecular data. The mitochondrial protein coding gene cytochrome c oxidase subunit I (COI) was chosen for sequencing; 1341 characters were obtained for 24 ingroup taxa and these were analyzed with parsimony. The monophyly of the genus Cheilosia was well supported. Current taxonomic division of Cheilosia into two subgenera (sg. Nigrocheilosia and sg. Neocheilosia) and most nonformalized species groups based on morphology were supported by the monophyletic groups identified in the molecular analysis. The phylogenetic informativeness of COI in resolving the subtribal relationships within the tribe Cheilosiini remains ambiguous. PMID- 10837154 TI - Nuclear genes resolve mesozoic-aged divergences in the insect order Lepidoptera. AB - Compared to the number of genes available for study of both younger and older divergences, few genes have yet been identified that can strongly resolve phylogenetic splits of Mesozoic age ( approximately 65-250 mya). Thus, reconstruction of Mesozoic-age phylogenies, exemplified by basal divergences within the major orders of holometabolous insects, is likely to be especially dependent on combining multiple lines of evidence. This study tests the potential of the 18S ribosomal RNA gene for reconstructing Mesozoic-aged divergences within the insect order Lepidoptera and its ability when combined with a second, previously analyzed nuclear gene (phosphoenolpyruvate carboxykinase, PEPCK) to strongly resolve these relationships. 18S sequences were obtained for 21 taxa, representing major clades of Lepidoptera plus outgroups from the other "panorpoid orders. A well-corroborated morphology-based "test phylogeny was used to evaluate the effects of partitioning the 18S gene according to variable versus conserved domains, paired versus unpaired sites in the secondary structure, and transition versus transversion substitutions. Likelihood and unweighted parsimony analyses of the 18S data recover the "test phylogeny" almost completely, with no improvement of agreement or support provided by any form of weighting or partitioning. No conflict in signal between 18S and PEPCK was detected by the partition homogeneity test. Combined parsimony analysis yielded strong bootstrap support for nearly all relationships, much higher than for either gene alone, thereby also providing strong evidence on several hypotheses about the early evolution of lepidopteran-plant interactions. These genes in combination may be widely useful for resolving insect divergences of comparable age. PMID- 10837155 TI - Mitochondrial DNA variation and the evolutionary history of cryptic Gammarus fossarum types. AB - The evolutionary history of the cryptic Gammarus fossarum species complex (Crustacea, Amphipoda) in Central Europe was approached by investigating the genetic variation in populations of a natural contact zone. Nucleotide sequence variation of a 395-bp segment of the mitochondrial 16S rRNA gene was compared to that of six nuclear allozyme loci. Three major mtDNA lineages were found, the eastern clade being consistent with the former allozyme type A. The two western clades (types B and C) were not distinguished previously. Strong sequence divergence and correlation with nuclear genetic isolation in syntopic populations, however, justifies the specific status of the three G. fossarum types. The common speciation event is believed to be very old (Miocene). The within-type mtDNA variation is probably molded by the ice ages, with type B populations being most affected. Moreover, the patch-like distribution of mtDNA type B lineages in an area near the contact zone corroborates the hypothesis of a recent colonization. PMID- 10837156 TI - Historical biogeography of the Western Rattlesnake (Serpentes: viperidae: Crotalus viridis), inferred from mitochondrial DNA sequence information. AB - We infer the phylogeography of the Western Rattlesnake (Crotalus viridis) using phylogenetic analysis of mitochondrial DNA sequences from 1345 bp of the genes for cytochrome b and NADH dehydrogenase subunit 4. Two main clades are revealed: one includes populations from east and south of the Rocky Mountains (conventionally referred to as Crotalus viridis viridis and C. v. nuntius), and the other consists of populations west of the Rocky Mountains. Within the western clade, a population from southern Arizona (C. v. cerberus) represents the sister taxon to the remaining western populations. The conventional subspecies recognized in this species do not fully correspond to the phylogenetic pattern, and a review of the systematic status of several populations is needed. Our data allow the inferences that small body size evolved twice and that the ability of one population (C. v. concolor) to secrete highly lethal toxins related to Mojave toxin arose within the complex. Our phylogeny should represent the basis for further studies on the causes of geographical variation in this complex. PMID- 10837157 TI - Evolutionary change of codon usage for the histone gene family in Drosophila melanogaster and Drosophila hydei. AB - The nucleotide divergence in the protein-coding region for replication-dependent and replication-independent histone 3 and 4 genes of Drosophila melanogaster and Drosophila hydei occurred mostly at the synonymous site. Therefore, the pattern of codon usage was analyzed in the two species, considering the genomic codon bias, which is proposed for estimating the genomic composition pressure in the protein-coding regions. The results indicated that the codon usage in the histone gene family could be explained mostly by the genomic codon bias. However, biases for Ala and Arg were commonly observed for the histone 3 and histone 4 gene families, and biases for Ser, Leu, and Glu were observed in a gene-specific manner. This suggests that both genomic codon bias and gene- or codon-specific bias are responsible for the nucleotide differentiation in the protein-coding region of the histone genes. PMID- 10837158 TI - Molecular systematics, zoogeography, and evolutionary ecology of the atlantic parrotfish genus Sparisoma. AB - Parrotfishes of the genus Sparisoma (Scaridae) are ecologically important tropical reef fishes restricted to the Atlantic Ocean. We investigated phylogenetic relationships among the eight extant species within this genus using mitochondrially encoded 12S and 16S ribosomal genes. Our molecular data support the view that (i) Sparisoma originated approximately 14-35 million years ago (mya), probably in the tropical western Atlantic, off Brazil; (ii) there have been at least four discrete bouts of cladogenesis within the genus, with the most recent one ( approximately 2.8-5.6 mya) involving four events in both the east and the west Atlantic and across the Atlantic; and (iii) the genus invaded the eastern Atlantic on two different occasions, probably by at least two different routes. The data also offer support for Bellwood's ideas concerning the evolutionary changes in adult feeding patterns and habitat use within Scarids. Specifically, they support the evolutionary position of the ecological traits of Sparisoma as intermediate within the family. PMID- 10837159 TI - Evolution of parental care and ovulation behavior in oysters. AB - Approximately half of all living oysters brood offspring in the inhalant chamber of their mantle cavities; the remainder are broadcast spawners which do not engage in parental care of young. Ostreid ovulation involves a complex behavioral sequence that results in the countercurrent passage of newly spawned eggs through the gills (ctenidia) and into the inhalant chamber. We constructed molecular and combined-evidence phylogenetic trees to test hypotheses concerning the directionality of parental care evolution, and the evolutionary significance of the trans-ctenidial ovulation pathway, in the Ostreidae. Representatives of all three ostreid subfamilies, together with gryphaeid and nonostreoidean pterioid outgroups, were sequenced for a 941-nucleotide fragment of the 28S ribosomal gene. Our phylogenetic analyses indicate that (1) the Ostreidae are robustly monophyletic, (2) broadcast spawning and larval planktotrophy are ancestral ostreid traits, (3) trans-ctenidial ovulation predates the evolution of parental care in ostreid lineages, and (4) brooding originated once in the common ancestor of the Ostreinae/Lophinae, involved a modification of the final behavioral step in the ancestral ovulation pathway, and has been retained in all descendent lineages. Our data permit an independent test of fossil-based ostreid phylogenetic hypotheses and provide novel insights into oyster evolution and systematics. PMID- 10837160 TI - Molecular phylogenetic examination of the delphinoidea trichotomy: congruent evidence from three nuclear loci indicates that porpoises (Phocoenidae) share a more recent common ancestry with white whales (Monodontidae) than they do with true dolphins (Delphinidae). AB - Porpoises (Phocoenidae), dolphins (Delphinidae), and the two species of Monodontidae (beluga and narwhal) together constitute the superfamily Delphinoidea. Although there is extensive evidence supporting the monophyly of this superfamily, previous studies involving morphology, as well as sequence analysis of mitochondrial genes, have failed to yield a clear picture of the relative relationships within the group. Here we present the first examination of this issue from the perspective of single-copy nuclear genes at the DNA sequence level. The data involve three such loci: von Willebrand factor (vWF), interphotoreceptor retinoid binding protein (IRBP), and lactalbumin. The vWF and IRBP data sets consist of protein-coding fragments, whereas the sequenced lactalbumin fragment is predominately intronic. All phylogenetic analyses involving at least one representative from each of the three Delphinoidea families congruently support a beluga/porpoise clade. The levels of sequence divergence for most of these data appear to roughly concur with a paleontological date for the radiation of the Delphinoidea at 11-15 MYA but, in agreement with mitochondrial DNA sequence analyses, suggest that the extant major groups of cetaceans radiated approximately 25 MYA, 10 million years later than inferred from paleontological data. PMID- 10837161 TI - Phylogeography of mitochondrial DNA variation in brown bears and polar bears. AB - We analyzed 286 nucleotides of the middle portion of the mitochondrial cytochrome b gene of 61 brown bears from three locations in Alaska and 55 polar bears from Arctic Canada and Arctic Siberia to test our earlier observations of paraphyly between polar bears and brown bears as well as to test the extreme uniqueness of mitochondrial DNA types of brown bears on Admiralty, Baranof, and Chichagof (ABC) islands of southeastern Alaska. We also investigated the phylogeography of brown bears of Alaska's Kenai Peninsula in relation to other Alaskan brown bears because the former are being threatened by increased human development. We predicted that: (1) mtDNA paraphyly between brown bears and polar bears would be upheld, (2) the mtDNA uniqueness of brown bears of the ABC islands would be upheld, and (3) brown bears of the Kenai Peninsula would belong to either clade II or clade III of brown bears of our earlier studies of mtDNA. All of our predictions were upheld through the analysis of these additional samples. PMID- 10837162 TI - Re: Spectral analysis of echinoderm small subunit ribosomal RNA gene sequence data. PMID- 10837163 TI - Microwave Spectrum of 1-Cyano-3-fluoro-but-1-ene. AB - The rotational spectrum of the 1-cyano-3-fluoro-but-1-ene has been recorded with a pulsed-nozzle microwave Fourier transform spectrometer over the range 6-20 GHz. The frequencies were fitted to the Hamiltonian of Watson (A-reduction, I(r) representation). The resulting rotational constants are A = 7493.404(1) MHz, B = 1211.9831(2) MHz, and C = 1096.0908(1) MHz. By comparing the experimental rotational constants with those obtained by ab initio calculations, we found without ambiguity that the stable conformation for the molecule is the one with the fluorine atom lying in the C&bond;CCN plane (CF-eclipsed conformer). Copyright 2000 Academic Press. PMID- 10837164 TI - The High-Resolution FTIR Spectrum and Equilibrium Structure of Chloro(sulphido)boron. AB - The FTIR spectrum of chloro(sulphido)boron, Cl-B&bond;S, has been recorded at 0.0035 cm(-1) resolution. The nu(3) bands of the isotopomers (35)Cl(11)B(32)S, (37)Cl(11)B(32)S, (35)Cl(11)B(34)S, (37)Cl(11)B(34)S, (35)Cl(10)B(32)S, (37)Cl(10)B(32)S, and (35)Cl(10)B(34)S have been analyzed to yield effective rotational and centrifugal distortion constants for the ground and a number of excited vibrational states. For the (35)Cl(11)B(32)S isotopomer the nu(1) band has also been analyzed. The rotational constants have been used to determine equilibrium bond lengths of 168.42(8) and 160.14(8) pm for the Cl-B and B-S bonds, respectively. Copyright 2000 Academic Press. PMID- 10837165 TI - Rhinoviruses infect the lower airways. AB - Rhinoviruses are the major cause of the common cold and a trigger of acute asthma exacerbations. Whether these exacerbations result from direct infection of the lower airway or from indirect mechanisms consequent on infection of the upper airway alone is currently unknown. Lower respiratory infection was investigated in vitro by exposing primary human bronchial epithelial cells to rhinoviruses and in vivo after experimental upper respiratory infection of human volunteers. Bronchial infection was confirmed by both approaches. Furthermore, rhinoviruses induced production of interleukin-6, -8, and -16 and RANTES and were cytotoxic to cultured respiratory epithelium. This evidence strongly supports a direct lower respiratory epithelial reaction as the initial event in the induction of rhinovirus-mediated asthma exacerbations. The frequency of infection and the nature of the inflammatory response observed are similar to those of the upper respiratory tract, suggesting that rhinovirus infections may be one of the most important causes of lower in addition to upper respiratory disease. PMID- 10837166 TI - Rhinovirus-induced oxidative stress and interleukin-8 elaboration involves p47 phox but is independent of attachment to intercellular adhesion molecule-1 and viral replication. AB - Virus-induced elaboration of proinflammatory cytokines is mediated by virus induced oxidative stress. The purpose of these studies was to determine the source of the virus-induced oxidative stress. Inhibition of viral replication with antibody to intercellular adhesion molecule-1 had no effect on virus-induced oxidative stress or interleukin-8 (IL-8) response (597+/-88 vs. 668+/-78 pg/mL in control cells). Treatment of cells with diphenylene iodonium inhibited virus induced oxidative stress and IL-8 elaboration, but allopurinol, ibuprofen, and rotenone had no effect. Studies in cell lines produced from a patient with gp91 phox deficiency revealed normal responses. In contrast, the oxidative response was decreased and the IL-8 concentration was 227+/-36 pg/mL in cells from a patient with p47-phox deficiency, compared with 664+/-48 pg/mL in control cells. These studies suggest that the stimulation of reactive oxygen species by viral challenge occurs at the cell surface even in the absence of viral replication and involves a flavoprotein that may act in concert with p47-phox. PMID- 10837167 TI - Circulation patterns of group A and B human respiratory syncytial virus genotypes in 5 communities in North America. AB - Human respiratory syncytial virus (HRSV) is a major cause of serious lower respiratory tract illness in infants, young children, and the elderly. To characterize the circulation patterns of HRSV strains, nucleotide sequencing of the C-terminal region of the G protein gene was performed on 34-53 isolates obtained from 5 communities during 1 epidemic year, representing distinct geographical locations in North America. Phylogenetic analysis revealed that 5-7 HRSV genotypes, including 1 or 2 predominant strains, circulated in each community. The patterns of genotypes were distinct between communities, and less diversity was seen between strains of the same genotype within than between communities. These findings are consistent with HRSV outbreaks' being community based in nature, although transmission of viruses between communities may occur. Several strains are probably introduced or circulate endemically in communities each year, and local factors-possibly immunity induced by previous years' strains determine which strains predominate during an HRSV season. PMID- 10837168 TI - Epidemiology of primary varicella and herpes zoster hospitalizations: the pre varicella vaccine era. AB - To determine the epidemiology and costs of hospitalization with primary varicella and herpes zoster in the prevaccine era and the usefulness of hospital discharge data to determine the population impact of vaccination on these conditions, statewide hospital discharge data in Connecticut from 1986 to 1995 were analyzed. Annual hospitalizations for herpes zoster were 4-fold higher than for primary varicella (16.1 vs. 4.1/100,000). Overall, 69% and 83%, respectively, had no underlying immunosuppressive conditions. Regarding primary varicella, 53% of patients were aged <15 years, there was a marked winter-spring seasonality, and Hispanics and blacks were 4.1 and 2.6 times more likely than whites to be hospitalized. Regarding herpes zoster, 66.9% of patients were aged >64 years, and there was no seasonality. The mean patient charges in 1995 were $12,819 for primary varicella and $15,583 for herpes zoster. Analysis of population-based hospital discharge data is a feasible means of monitoring the impact of varicella immunization on severe morbidity due to primary varicella and herpes zoster. PMID- 10837169 TI - Combination treatment with famciclovir and a topical corticosteroid gel versus famciclovir alone for experimental ultraviolet radiation-induced herpes simplex labialis: a pilot study. AB - To investigate the efficacy of corticosteroids for the treatment of herpes labialis, we compared famciclovir (Famvir, 500 mg 3x/day po [per os] for 5 days) and topical fluocinonide (0.05% Lidex Gel 3x/day for 5 days) with famciclovir and topical vehicle control for experimental ultraviolet radiation-induced herpetic recurrences. We irradiated 49 volunteers, and 29 (60%) of 48 developed signs or symptoms of a recurrence. They self-initiated treatment, and we were able to evaluate them. There was a trend in the combination group toward more aborted lesions, compared with those who received antiviral therapy alone (7 [41%] of 17 vs. 1 [8%] of 12; P=.09). Combination therapy significantly reduced the median maximum lesion size (48 vs. 162 mm(2); P=.02) and the number of patients who experienced lesion pain (10 [59%] of 17 vs. 12 [100%] of 12; P=.02). Adverse events were minimal. Corticosteroids in combination with an antiviral agent may be safe and beneficial for episodic treatment of herpes labialis. Larger studies are needed to confirm these findings. PMID- 10837170 TI - Comparison of human papillomavirus types 16, 18, and 6 capsid antibody responses following incident infection. AB - The relationship between human papillomavirus (HPV) DNA in the genital mucosa and serum IgG to HPV-16, -18, and -6 was studied in a cohort of 588 college women. Among women with incident HPV infections, 59.5%, 54.1%, and 68.8% seroconverted for HPV-16, -18, or -6, respectively, within 18 months of detecting the corresponding HPV DNA. Transient HPV DNA was associated with a failure to seroconvert following incident HPV infection; however, some women with persistent HPV DNA never seroconverted. Antibody responses to each type were heterogeneous, but several type-specific differences were found: seroconversion for HPV-16 occurred most frequently between 6 and 12 months of DNA detection, but seroconversion for HPV-6 coincided with DNA detection. Additionally, antibody responses to HPV-16 and -18 were significantly more likely to persist during follow-up than were antibodies to HPV-6. PMID- 10837171 TI - Virologic analysis of non-B, non-C hepatocellular carcinoma in Japan: frequent involvement of hepatitis B virus. AB - Serum and liver tissues from hepatitis B surface antigen-negative/anti-hepatitis C virus (HCV)-negative (non-B, non-C) hepatocellular carcinoma (HCC) patients in Japan were examined for the presence of hepatitis B virus (HBV), HCV, and TT virus (TTV) by polymerase chain reaction. The studies evaluated the contribution of these viruses to pathogenesis of HCC. HBV DNA was detected in the sera of 20 (47.6%) of 42 non-B, non-C HCC patients, which was significantly higher than in age-matched controls without liver disease (P<.001). In 8 of 12 patients with liver tissues available, HBV DNA was detected in cancerous and adjacent noncancerous liver tissues. No HCV RNA was detected. The positivity for TTV DNA was not significantly different between HCC patients and controls. These results indicate that HBV is associated with a substantial proportion of non-B, non-C HCC cases in Japan. The role of HBV in hepatocarcinogenesis in such patients needs to be clarified. PMID- 10837172 TI - Increased level of interferon-alpha in blood of patients with insulin-dependent diabetes mellitus: relationship with coxsackievirus B infection. AB - The activation of the interferon (IFN)-alpha system and its relationship with coxsackievirus B (CVB) infection has been analyzed in 56 patients with insulin dependent diabetes mellitus (IDDM; 25 children and 31 adults). Elevated levels of IFN-alpha were found in plasma of 70% of patients (39/56), and a positive detection of IFN-alpha mRNA in blood cells by reverse transcriptase-polymerase chain reaction (RT-PCR) was observed in 75% of patients (42/56). Enterovirus (EV) RNA assayed by seminested RT-PCR was detected in the blood of 50% of IFN-alpha positive patients but not in any IFN-alpha-negative patients. The results of genotype analysis of amplified EV RNA sequences (5 CVB2, 8 CVB3, and 8 CVB4) were concordant with the results of CVB-neutralization tests. The comparison between IFN-alpha, EV RNA, and serology suggested that the proportion of CVB infection associated with IFN-alpha positivity might be higher than is predicted from the investigation of EV RNA. Together, the results suggest that, in a majority of cases, a CVB infection is associated with clinical IDDM. PMID- 10837173 TI - Interaction of human immunodeficiency virus type 1 and human herpesvirus type 8 infections on the incidence of Kaposi's sarcoma. AB - To determine Kaposi's sarcoma (KS) risk related to timing of human immunodeficiency virus type 1 (HIV-1) and human herpesvirus type 8 (HHV-8) infections, stored longitudinal sera from 400 homosexual men with known dates of HIV-1 seroconversion (+/-4.5 months) were tested for HHV-8 antibody. Times from HHV-8 seroconversion to KS were compared for the 69 men who became infected with HHV-8 after acquiring HIV-1 to the 182 men who were HHV-8 seropositive before their HIV-1 infection. None developed KS before coinfection. HHV-8 seroconversion after HIV-1 infection increased the risk of KS (risk ratio, 2.55; 95% confidence interval, 1.06-6.10) compared with those infected with HHV-8 before HIV-1. The KS hazards in HHV-8-infected men increased by 60% (P<.001) for each year of HIV-1 infection. Faster CD4 cell loss and higher HIV-1 RNA levels significantly predicted KS. The quicker development of KS in men acquiring HHV-8 after HIV-1 and its association with CD4 slope argues that KS is more likely if HHV-8 infection occurs in an immunocompromised person. PMID- 10837174 TI - Correlation between human immunodeficiency virus type 1 RNA levels in the female genital tract and immune activation associated with ulceration of the cervix. AB - To address the hypothesis that local immune activation resulting from genital ulceration enhances human immunodeficiency virus type 1 (HIV-1) replication and shedding into the genital tract, paired plasma and cervicovaginal lavage (CVL) samples were obtained from 12 HIV-infected women before and after treatment of cervical intraepithelial lesions. Two weeks after treatment, inflammation and ulceration of the cervix were accompanied by major increases in mean concentrations of HIV-1 RNA (200-fold), tumor necrosis factor-alpha, interleukin 6, and soluble markers shed by activated lymphocytes and macrophages (sCD25 and sCD14, respectively) in CVL samples (P<.01 for each), but not plasma. Strong temporal and quantitative correlations were observed between concentrations of immunological markers and HIV-1 load in this compartment during a 10-week follow up. Furthermore, in the presence of genital ulceration, HIV-1 in CVL samples was more readily captured by antibodies directed against virion-associated HLA-DR, a marker of host-cell activation, compared with virus in plasma. We suggest that local immune activation increases HIV-1 load in genital secretions, potentially increasing the risk of HIV-1 transmission. PMID- 10837175 TI - Lack of association between human immunodeficiency virus type 1 antibody in cervicovaginal lavage fluid and plasma and perinatal transmission, in Thailand. AB - To determine the association between human immunodeficiency virus type 1 (HIV) specific antibody and RNA levels in cervicovaginal lavage (CVL) samples and plasma, zidovudine treatment, and perinatal transmission, HIV subtype E gp160 specific IgG and IgA were serially measured in a subset of 74 HIV-infected women in a placebo-controlled trial of zidovudine, beginning at 36 weeks of gestation. HIV IgG was detected in 100% of plasma and 97% of CVL samples; HIV IgA was consistently detected in 62% of plasma and 31% of CVL samples. Antibody titers in CVL samples correlated better with the RNA level in CVL samples than with plasma antibody titers. Zidovudine did not affect antibody titers. Perinatal HIV transmission was not associated with antibody in CVL samples or plasma. HIV specific antibody is present in the cervicovaginal canal of HIV-infected pregnant women; its correlation with the RNA level in CVL fluid suggests local antibody production. However, there was no evidence that these antibodies protected against perinatal HIV transmission. PMID- 10837176 TI - Systemic inflammation in hemorrhagic fever with renal syndrome correlates with hypotension and thrombocytopenia but not with renal injury. AB - Systemic inflammation is common in patients with nephropathia epidemica (NE), a European form of hemorrhagic fever. Markers of inflammation were studied in a patient with NE with respiratory insufficiency (patient 1), 18 other patients with NE, and 13 patients with a viral infectious disease other than NE. Neutrophil and monocyte CD11b expression levels, determined by flow cytometry; soluble interleukin (IL)-2 receptor (sIL-2R), IL-6, and IL-8 concentrations, determined by means of Immulite; and soluble E-selectin, determined by ELISA, were higher in patients with NE than in healthy subjects. The findings were not specific for NE and did not correlate with serum creatinine levels, but the findings correlated inversely with mean arterial pressure (sIL-2R and monocyte CD11b expression) and minimum platelet count (sIL-2R, IL-6, neutrophil, and monocyte CD11b expression). Monocyte CD11b expression in patient 1 was extremely high, suggesting that monocytes may contribute to development of lung injury. Severity of inflammation in patients with NE is related to hypotension and platelet consumption but not to renal injury. PMID- 10837177 TI - Molecular epidemiology survey of penicillin-susceptible and -resistant Streptococcus pneumoniae recovered from patients with meningitis in France. AB - The genetic diversity of Streptococcus pneumoniae isolates (n=291) recovered from cerebrospinal fluid of patients with meningitis in France was investigated by restriction fragment length polymorphism analysis of the ribosomal RNA gene regions and of the pbp2b and 2x genes. Statistical analysis of the data by factorial analysis of correspondence established the following: penicillin susceptible isolates had a high level of genetic diversity, especially those belonging to serogroups frequently associated with carriage; capsular serotype switches could occur among penicillin-susceptible and -resistant isolates; and the mechanisms of acquired penicillin resistance were clearly distinct in isolates with penicillin minimum inhibitory concentration (MIC) values <1 mg/L and isolates with penicillin MIC values >/=1 mg/L. Thus, an increase in the penicillin MIC for a given strain, from intermediate to high-level resistance would be a rare event. PMID- 10837178 TI - Low cord blood type 14 pneumococcal IgG1 but not IgG2 antibody predicts early infant otitis media. AB - Type-specific IgG1 and IgG2 antibodies to Streptococcus pneumoniae capsular polysaccharides 14 and 19F were measured in cord blood samples from 425 neonates, to determine which antibody subclass was most strongly associated with otitis media (OM) during the first 6 months of life (early OM). Early OM was significantly associated with type 14 IgG1 antibody in the lowest antibody quartile (P=.055) but not with type 19F IgG1 antibody or with either IgG2 antibody. IgG1 and IgG2 antibodies were significantly intercorrelated for type 14 (r=.52, P<.001) and type 19F (r=.38, P<.001). Multivariate analysis revealed that having type 14 IgG1 antibody in the lowest quartile, child care attendance, and sibling and maternal OM history were independent risk factors for early OM. Although type-specific pneumococcal IgG2 antibody concentrations were significantly higher than IgG1 concentrations, IgG2 antibodies apparently are not protective against OM during early infancy. PMID- 10837179 TI - Sequential colonization by Streptococcus pneumoniae of healthy children living in an orphanage. AB - A prospective study of nasopharyngeal colonization by Streptococcus pneumoniae in the exceptional conditions of a closed community of abandoned children was done over a 1-year period; 71 children (age <24 months) were studied monthly. S. pneumoniae was isolated from 58 (81.7%), and 94.5% of the 111 isolates were resistant to penicillin. The mean rate of carriage was estimated at 57.4%, ranging from 42.8% to 70.4%. Children were sequentially colonized by a mean of 3 different isolates. The mean duration of carriage for a given isolate was approximately 2.2 months. Serotyping and molecular typing by pulsed-field gel electrophoresis showed that children were colonized by a limited number of clones belonging to only 4 serotypes and 4 pulsotypes. These clones rapidly spread in the community and colonized the children in waves, with a rapid turnover of S. pneumoniae isolates, facilitated by close contact between children. PMID- 10837180 TI - Cell-mediated immunity and antibody responses to Bordetella pertussis antigens in children with a history of pertussis infection and in recipients of an acellular pertussis vaccine. AB - Cell-mediated immunity (CMI) and antibody responses to Bordetella pertussis antigens were assessed 4-6 years after primary infant immunization with diphtheria-tetanus tricomponent acellular pertussis (DTaP) or diphtheria-tetanus (DT) vaccine in a country with high endemicity of B. pertussis infection. CMI to the B. pertussis antigens (especially to the pertussis toxin [PT]) was more frequent and/or intense in DTaP than in DT recipients. No lymphoproliferation differences were found between those with and without a history of pertussis although the DT recipients produced very little interferon-gamma after antigen (particularly PT and filamentous hemagglutinin [FHA]) stimulation. In contrast, seropositivity to PT, but not to pertactin or FHA, was more frequent in DT recipients with history of pertussis than in all other subjects. Thus, years after disease or vaccination, CMI response to PT or circulating PT antibodies appears to be the main distinctive feature of pertussis-protected DTaP recipients or pertussis-affected DT recipients. PMID- 10837181 TI - Biodistribution and genetic stability of the novel antitumor agent VNP20009, a genetically modified strain of Salmonella typhimurium. AB - VNP20009 is a genetically modified strain of Salmonella typhimurium possessing an excellent safety profile, including genetically stable attenuated virulence (a deletion in the purI gene), reduction of septic shock potential (a deletion in the msbB gene), and antibiotic susceptibility. VNP20009 is genetically stable after multiple generations in vitro and in vivo. In mice, VNP20009 is rapidly cleared from the blood from a peak level of 1x104 cfu/mL to undetectable levels in 24 h. In tumor-bearing mice, VNP20009 accumulates preferentially in tumors over livers at a ratio of 1000&rcolon;1. In nonhuman primates, VNP20009 was also rapidly cleared from the blood, from a peak level of 1.0x106 cfu/mL to undetectable levels in 24 h. VNP20009 was detected in the liver, spleen, and bone marrow of monkeys; the amount decreased over time, and VNP20009 was cleared from all organs by day 41; no VNP20009 could be detected in the urine or feces of the monkeys. VNP20009 is genetically stable after many generations of growth (>140) both in vitro and in vivo. PMID- 10837182 TI - Liposomal amphotericin B and amphotericin B-deoxycholate show different immunoregulatory effects on human peripheral blood mononuclear cells. AB - Conventional preparations of amphotericin B (AmB) at established therapeutic doses are known to increase nonspecific immune responses. It remains to be established whether higher doses of the less toxic liposomal preparation of AmB maintains a beneficial effect on the immune response to fungal infections. Examination of the effect of treatment of human peripheral blood mononuclear cells from healthy subjects with various doses of both liposomal AmB (L-AmB) and deoxycholate AmB (d-AmB) on proliferation, cell viability, and percentage of apoptosis demonstrated that, although both L-AmB and d-AmB at low doses significantly increased nonspecific proliferative responses, L-AmB, but not d AmB, treatment maintained this beneficial effect at higher doses. High doses of d AmB, but not L-AmB, resulted in significantly decreased cell viability and increased apoptosis. This study provides further evidence in healthy human subjects for choosing L-AmB over conventional preparations in the clinical treatment of fungal infections requiring systemic high-dose treatment with AmB. PMID- 10837183 TI - Murine CD4+ T lymphocyte subsets and host defense against Pneumocystis carinii. AB - The recruitment of specific subsets of CD4(+) T lymphocytes to the lungs in response to Pneumocystis carinii was investigated. For mice inoculated with P. carinii, an ELISPOT assay was used to calculate the numbers of lymph node and lung tissue CD4(+) cells that secreted interferon (IFN)-gamma (Th1 cytokine) and interleukin (IL)-4 (Th2 cytokine) after concanavalin A stimulation. An ELISA was used to assay culture supernatants for cytokine concentrations. Precursor frequency of both IFN-gamma- and IL-4-secreting cells was increased in lymph nodes at 1 week, whereas increases in Th1 and Th2 cells in lung tissue were delayed 3 weeks before declining. The frequency of IL-4-secreting cells always was greater than the frequency of IFN-gamma secreting cells. These results demonstrate an early T lymphocyte response in draining lymph nodes, followed by later recruitment of Th1 and Th2 lymphocytes into lung tissue. The overall CD4(+) T cell response to P. carinii involves both Th1 and Th2 subsets, but the response is Th2 dominant in both lymph node and lung tissue. PMID- 10837184 TI - Discrimination between patients with acquired toxoplasmosis and congenital toxoplasmosis on the basis of the immune response to parasite antigens. AB - Many persons infected with Toxoplasma gondii develop ocular lesions. Immunologic parameters in the response to T. gondii were evaluated in infected persons with and without ocular lesions and in noninfected controls. Subjects were divided into groups on the basis of presence of serum antibodies to T. gondii, presence of ocular lesions, and clinical history. Production of interleukin-2 and interferon-gamma by peripheral blood mononuclear cells from patients with probable congenital toxoplasmosis was decreased, compared with that in persons with presumed acquired infection. Cell proliferation and delayed-type skin reaction induced by soluble toxoplasma tachyzoite antigen followed the same pattern. Asymptomatic persons showed high levels of interleukin-12 and interferon gamma, whereas persons with ocular lesions had high interleukin-1 and tumor necrosis factor-alpha responses toward soluble toxoplasma tachyzoite antigen. These data suggest that patients with ocular disease due to congenital infection show tolerance toward the parasite. Furthermore, susceptibility to ocular lesions after acquired toxoplasmosis is associated with high levels of interleukin-1 and tumor necrosis factor-alpha, whereas resistance is associated with high levels of interleukin-12 and interferon-gamma. PMID- 10837185 TI - Molecular evidence of greater selective pressure for drug resistance exerted by the long-acting antifolate Pyrimethamine/Sulfadoxine compared with the shorter acting chlorproguanil/dapsone on Kenyan Plasmodium falciparum. AB - Pyrimethamine (PM) plus sulfadoxine (SD) is the last remaining affordable drug for treating uncomplicated malaria in Africa. The selective pressure exerted by the slowly eliminated combination PM/SD was compared with that exerted by the more rapidly eliminated combination chlorproguanil/dapsone (CPG/Dap) on Kenyan Plasmodium falciparum. Point mutations were analyzed in dihydrofolate reductase and dihydropteroate synthase and in the genetic diversity of 3 genes in isolates collected before and after CPG/Dap and PM/SD treatments. PM/SD was associated strongly with the disappearance of fully drug-sensitive parasites and with a significant increase in the prevalence of resistant parasites in subsequent parasitemias. However, this was not a characteristic of treatment with CPG/Dap. Moreover, most of the patients who returned with recrudescent infections were in the PM/SD-treated group. The data predict a longer useful therapeutic life for CPG/Dap than for PM/SD, and, thus, CPG/Dap is a preferable alternative for treatment of chloroquine-resistant falciparum malaria in sub-Saharan Africa. PMID- 10837186 TI - Members of the family caliciviridae (Norwalk virus and Sapporo virus) are the most prevalent cause of gastroenteritis outbreaks among infants in Japan. AB - Norwalk virus (NV) and Sapporo virus (SV) were approved as type species of the genus Norwalk-like viruses and the genus Sapporo-like viruses, respectively, within the family Caliciviridae. To clarify the importance of NV and SV as causes of gastroenteritis outbreaks in infants, stool samples obtained from 36 outbreaks of nonbacterial gastroenteritis that occurred during 1976-1995 in an infant home in Sapporo, Japan, were examined for diarrhea viruses using electron microscopy, enzyme immunoassays, reverse transcriptase-polymerase chain reaction (PCR), and sequencing of the PCR products. NV and SV were associated with 15 (42%) of the 36 outbreaks and were more prevalent than rotavirus (RV) A, which was associated with 10 (28%) of the 36 outbreaks. Our data indicate that NV and SV were the most common cause of outbreaks of viral gastroenteritis in infants and were indeed more prevalent than RV-A in Sapporo, Japan, during 1976-1995. PMID- 10837187 TI - Liver damage and kinetics of hepatitis C virus and human immunodeficiency virus replication during the early phases of combination antiretroviral treatment. AB - In order to assess the relationship between human immunodeficiency virus (HIV) RNA, hepatitis C virus (HCV) RNA, CD4, CD8, and liver enzymes during combination antiretroviral therapy, these parameters were measured in 12 HIV-HCV-coinfected patients (who were naive for antiretrovirals) on the day before and 3, 7, 14, 28, 56, and 84 days after initiating the following treatments: stavudine and lamivudine in all patients, indinavir in 6 patients, and nevirapine in 6 patients. HIV RNA declined rapidly, CD4 cells increased slowly, and CD8 cells and liver enzymes were stable. HCV RNA showed a transient significant increase at days 14 and 21 (7.33+/-0.16 [mean +/- SE] and 7.29+/-0.2 log copies/mL vs. 7+/ 0.2 log copies/mL at baseline; P<.05). These changes were similar in both treatment groups. A 2-fold alanine aminotransferase increase was observed in 4 of 12 patients; 4 of 4 patients showed increased HCV RNA. The relationship between HCV RNA increase and HIV RNA decrease indicates virus-virus interference. An HCV RNA increase may cause significant liver damage only in a minority of patients. PMID- 10837188 TI - Central nervous system activation of the indoleamine-2,3-dioxygenase pathway in human T cell lymphotropic virus type I-associated myelopathy/tropical spastic paraparesis. AB - Human T cell lymphotropic virus type I (HTLV-I) is associated with a chronic neurologic disease called HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP). The potential mechanisms of HAM/TSP pathogenesis were assessed by examination of 2 pathways initiated by interferon-gamma, a predominant cytokine in HAM/TSP. Jamaican HAM/TSP patients (n=17) were compared with patients with other neurologic diseases (ONDs; n=13) with respect to cerebrospinal fluid levels of the following: neopterin; nitrite plus nitrate, a stable indicator of nitric oxide; and tryptophan and kynurenine, metabolites of the indoleamine-2,3-dioxygenase (IDO) pathway. HAM/TSP patients had significantly elevated levels of neopterin (P=.003) and kynurenine (P=.05) and a significantly decreased level of tryptophan (P=.003), compared with patients with ONDs. These results support immune activation within the central nervous system and activation of the IDO pathway. Thus, activation of the IDO pathway may play a role in HAM/TSP. PMID- 10837189 TI - In vitro reactivation of human immunodeficiency virus 1 from latently infected, resting CD4+ T cells after bacterial stimulation. AB - Microbial coinfections have been associated with transient bursts of human immunodeficiency virus (HIV) viremia in patients. In this study, we have investigated whether microbial coinfections can induce replication of HIV-1 in latently infected CD4(+) T cells derived from HIV-infected patients who are receiving highly active antiretroviral therapy and in whom plasma viremia is undetectable by sensitive assays. We demonstrate that supernatants from macrophages exposed to the bacterial product lipopolysaccharide can induce in vitro activation of HIV-1 from latently infected, resting CD4(+) T cells obtained from HIV-infected individuals. Depletion of proinflammatory cytokines from the supernatant markedly reduced-whereas depletion of ss chemokines increased--the ability of the supernatant to induce replication of HIV-1. Our results suggest that coinfection with microbial pathogens such as bacteria may induce viral replication in the latent viral reservoirs in vivo. PMID- 10837190 TI - High prevalence of Epstein-Barr virus type 2 among homosexual men is caused by sexual transmission. AB - To investigate whether Epstein-Barr virus (EBV) type 2 infection is highly prevalent among homosexual men, the prevalence of EBV type 2 was studied among homosexual and heterosexual white men who were at high and low risk for sexually transmitted diseases; these data were correlated with sexual behavior. The prevalence of EBV type 2 among homosexual men was significantly higher than it was among heterosexual men (39% vs. 6%). Among high-risk heterosexual men, prevalence was significantly higher than it was among low-risk heterosexual men (15% vs. 0). In univariate analyses, EBV type 2 infection in homosexual men was significantly associated with human immunodeficiency virus (HIV) seropositivity, increased numbers of intercourse partners, non-Dutch nationality, and human herpesvirus 8 seropositivity. In multivariate analyses, an independent association with EBV type 2 was observed only for HIV seropositivity and number of sex partners. These data support the conclusion that EBV type 2 infection is more prevalent among white homosexual men and is caused by sexual transmission. PMID- 10837191 TI - Predictive value of Epstein-Barr virus genome copy number and BZLF1 expression in blood lymphocytes of transplant recipients at risk for lymphoproliferative disease. AB - Epstein-Barr virus (EBV) genome numbers and RNA transcripts from the immediate early EBV gene BZLF1 were monitored by means of polymerase chain reaction in peripheral blood lymphocytes (PBLs) of 44 children who received liver transplants. The 2 tests were compared, using several parameters to assess their value as predictors of posttransplantation lymphoproliferative disease (PTLD). All patients were infected with EBV. BZLF1 mRNA was positive in 70% of patients, with highest expression in those with largest virus load. Four patients developed PTLD that could not be unequivocally diagnosed by any of the parameters considered alone. Sensitivity of EBV genome number (>/=40,000 EBV copies/10(5) PBLs) and BZLF1 mRNA (BZLF1:glyceraldehyde-3-phosphate-dehydrogenase ratio >/=0.5) was 100%. Specificity of each of the 2 tests alone (98% and 58%, respectively) improved (to 100% and 83%, respectively) when measurement of serum IgG level was included. Because decreased virus load, but not BZLF1 mRNA expression, accurately predicted favorable responses of PTLD to therapy, monitoring of EBV genome numbers alone appears sufficient in children with liver transplants. PMID- 10837192 TI - Resistance to antiviral drugs in herpes simplex virus infections among allogeneic stem cell transplant recipients: risk factors and prognostic significance. AB - Herpes simplex virus (HSV) infections in 75 allogeneic stem cell transplant recipients were analyzed. Sixteen patients developed HSV disease following transplantation. The risk factors were age, sex (females), unrelated donor graft, and graft-versus-host disease (GVHD) grade >/=2. Seven patients did not respond to acyclovir, and 3 patients failed to respond to foscarnet. Isolates from 4 patients developed resistance to acyclovir/penciclovir, and 3 patients had foscarnet-resistant isolates. The remaining 3 patients failed to respond to acyclovir, despite having sensitive isolates. All the isolates were sensitive to cidofovir, for which the IC(50) values correlated inversely with those for acyclovir (P=.01). The risk factors for clinical resistance to antiviral drugs were a GVHD grade >/=2 (P=.001) and the lack of ganciclovir prophylaxis (P=.01), with a higher nonrelapse mortality in the latter group (P<.0001). Clinical as well as in vitro resistance to antiviral drugs is common in patients with severe GVHD and is associated with a poor outcome. PMID- 10837193 TI - IgG subclass distribution of hepatitis B surface antigen antibodies induced in children with chronic hepatitis B infection after interferon-alpha therapy. AB - The IgG subclass distribution of antibody to hepatitis B surface antigen (anti HBs) was investigated in 19 children with chronic active hepatitis B infection who showed a complete serological seroconversion after interferon-alpha therapy. Determinations were done 6 and 12 months after treatment. Our results showed no selectivity in anti-HBs synthesis among IgG subclasses. All 4 IgG isotypes were involved in the response, with similar percentage contributions, on average, of IgG1 (35%), IgG3 (27%), and IgG4 (28%), followed by IgG2 (10%). IgG4 became the second most dominant isotype at the end of observation. These results are in contrast to those found after natural seroconversion, in which anti-HBs was highly restricted to neutralizing IgG1 and IgG3, with only a minor contribution from IgG2 and IgG4. It is postulated that analysis of the specific profiles of IgG subclasses may be of value for the estimation of the therapeutic efficacy of recombinant interferon-alpha used and may be helpful in choosing more-effective treatment. PMID- 10837194 TI - Monitoring the virus load can predict the emergence of drug-resistant hepatitis B virus strains in renal transplantation patients during lamivudine therapy. AB - The development of resistant hepatitis B virus (HBV) strains during lamivudine treatment has been described repeatedly. To investigate whether the development of such resistant HBV strains can be predicted in an early phase of therapy, the HBV loads of 11 renal transplantation patients were screened at 3-month intervals by a quantitative HBV polymerase chain reaction (PCR) assay. Lamivudine resistance was detected by sequence analysis. Five patients developed resistance to lamivudine in the 12-15-month follow-up period. In all of them, a virus load of 1x103 HBV DNA copies still was detectable after 3 months of therapy. This was statistically significantly different from those patients who did not develop lamivudine resistance within the observation period, all of whom had no HBV DNA detectable after 3 months of treatment (P=.0022). Thus, virus load testing by use of a sensitive PCR assay allows the early prediction of the emergence of lamivudine-resistant HBV strains. PMID- 10837195 TI - Granulocyte colony-stimulating factor receptors on granulocytes are down regulated after endotoxin administration to healthy humans. AB - Granulocyte colony-stimulating factor (G-CSF) is considered an important mediator of host defense against infection, and recombinant G-CSF is administered to patients with various infections. G-CSF binds to a specific receptor that is expressed on granulocytes and monocytes. To obtain insight about the regulation of the G-CSF receptor after an acute infectious challenge, 8 healthy subjects received an intravenous injection of lipopolysaccharide (LPS; 4 ng/kg), and receptor expression was determined on blood leukocytes by fluorescence-activated cell sorter analysis, both by measurement of saturation binding of recombinant G CSF and by use of an anti-G-CSF-receptor antibody. LPS induced a transient decrease in granulocyte, but not monocyte, G-CSF-receptor expression. In whole blood in vitro, not only LPS but also gram-positive stimuli and proinflammatory cytokines were capable of down-modulating the G-CSF receptor on granulocytes. Bacterial antigens down-regulate the G-CSF receptor at the surface of granulocytes, which may impair neutrophil functions important for antibacterial host defense. PMID- 10837196 TI - Human antibody response to longus type IV pilus and study of its prevalence among enterotoxigenic Escherichia coli in Bangladesh by using monoclonal antibodies. AB - Mouse monoclonal antibodies (MAbs) were derived against longus (CS20), a type IV pilus expressed by human enterotoxigenic Escherichia coli (ETEC). One MAb (ICA39) detected longus in 56 (8.5%) of 662 ETEC isolates obtained from a routine surveillance of diarrheal stools from children and adults. Five patients with diarrhea from whom longus-positive ETEC were isolated were also recruited. Of these 61 isolates, 50 were positive for other colonization factors (CFs; 61% for CFA/II and 21% for CFA/I), and 11 were negative for any of the other 8 CFs that were tested. They were either positive for the heat-stable enterotoxin (ST; n=29) or for the heat-labile enterotoxin (LT) and ST (n=32). All longus-positive ETEC were confirmed by polymerase chain reaction to harbor lngA, the longus structural pilin gene. Sera and/or fecal extracts from the patients reacted with the 22-kDa pilin polypeptide in immunoblots and ELISA. These studies show that longus is prevalent among ETEC in Bangladesh and that longus gives rise to IgA antibody responses in patients. PMID- 10837197 TI - A 5-year study of the seroepidemiology of Klebsiella pneumoniae: high prevalence of capsular serotype K1 in Taiwan and implication for vaccine efficacy. AB - Seroepidemiology of Klebsiella pneumoniae was determined for 1000 nonrepetitive K. pneumoniae isolates collected by a medical center in Taiwan during 1993-1997. Of these, 630 isolates (63%) were from community-acquired infections; the rest were from hospital-acquired infections. The isolates were serotyped according to capsular antigen by countercurrent immunoelectrophoresis. About 77% were typeable. Serotypes K1 and K2 accounted for 21.7% and 9.3% of the isolates, respectively, followed by K57 (5.1%), K54 (4.2%), K21 (3. 3%), and K16 (3%). The frequency of serotype K1 among bacteremic isolates (30.8%) far exceeded that reported by other investigators worldwide. Molecular typing of random K1 isolates by pulsed-field gel electrophoresis revealed several different pulsotypes, suggesting a nonclonal spread. This study indicates that a Klebsiella vaccine developed in Europe is not optimal for use in Taiwan because it does not contain the most predominant serotypes-K1, K54, and K57. PMID- 10837198 TI - Gonorrhea among men who have sex with men: outbreak caused by a single genotype of erythromycin-resistant Neisseria gonorrhoeae with a single-base pair deletion in the mtrR promoter region. AB - During 1995-1997, an outbreak of 66 cases of gonorrhea caused by an erythromycin resistant (Ery(r); MIC >/=1.0 microgram/mL) prototrophic (proto) auxotype IB-1 serovar of Neisseria gonorrhoeae occurred in King County, Washington; 65 cases involved men who have sex with men (MSM), which accounted for approximately 37% of infections among MSM during this period. Isolates from 19 of these 65 cases of infection were analyzed by DNA sequencing of the polymerase chain reaction amplified promoter region of the mtrR gene and by pulsed-field gel electrophoresis (PFGE) analysis of genomic DNA after NheI and SpeI digestion. Eighteen of the 19 isolates had a 1-bp A/T deletion in a 13-bp inverted repeat of the mtrR promoter region and shared a single PFGE type. Among MSM who provided data about sexual behavior, 37 (64%) of 58 MSM infected by the proto/IB-1 Ery(r) strain reported having >2 sex partners during the past 60 days, compared with 32 (30%) of 106 MSM infected by other strains (P<.001). This clonal outbreak of gonorrhea illustrates the ongoing need for behavioral preventive interventions among MSM. PMID- 10837199 TI - Conservation of Helicobacter pylori genotypes in different ethnic groups in Houston, Texas. AB - This study was concerned with whether the Helicobacter pylori strains circulating among ethnic groups living in the same region differ. The polymerase chain reactions to genotype (cagA, vacA, and iceA) H. pylori isolates from healthy volunteers from 4 ethnic groups (black, n=35; white Hispanic, n=31; whites, n=30; Vietnamese, n=29) residing in Houston were examined. The Vietnamese volunteers had the "East Asian"-type cagA 3' repeat region structure, and the others had the "non-Asian" type. The most common genotypes were delineated as follows: blacks and Hispanics, cagA+, vacA s1b-m1, and iceA2; whites, cagA+, vacA s1a-m2, and iceA2; and Vietnamese, cagA+, vacA s1c-m2, and iceA2. Two Hispanic families were also examined. H. pylori isolates from the children and their mothers had the same genotype and were different from those associated with the children's fathers or brothers-in-law. Conservation of an H. pylori genotype within ethnic groups over the course of generations will prove useful for epidemiological study of the coevolution of humans and H. pylori. PMID- 10837200 TI - Protection against Bordetella pertussis in mice in the absence of detectable circulating antibody: implications for long-term immunity in children. AB - Most vaccines used for humans work through humoral immunity, yet many appear to be protective even after specific circulating antibody levels have waned to undetectable levels. Furthermore, it has been difficult to define a serologic correlate of protection against a number of infectious diseases, including those caused by Bordetella pertussis. B. pertussis clearance in immunized mice has been shown to correlate with pertussis vaccine efficacy in children. This murine respiratory challenge model was used to demonstrate persistent vaccine-induced protection against B. pertussis in the absence of circulating antibody at the time of challenge. Whole-cell and acellular pertussis vaccines induced persistent memory T and B cells and anamnestic antibody responses after challenge. The findings suggest that immunologic memory is more significant in protection than is the induction of immediate antibody responses and imply that vaccinated children still may be protected against disease following the disappearance of specific serum IgG. PMID- 10837201 TI - Hepatocyte growth factor levels in cerebrospinal fluid: a comparison between acute bacterial and nonbacterial meningitis. AB - The organotrophic functions of the hepatocyte growth factor (HGF) have been the subject of several studies. In the more recent studies, this function has been reported in the brain. In the present study, we have measured the levels of HGF in cerebrospinal fluid (CSF) and sera from 78 patients divided into 6 different groups according to central nervous system (CNS) infection and control. Quantitative measurements of HGF in the CSF and serum were performed by an enzyme linked immunosorbent assay. Elevated values of CSF HGF were found in the patients with acute bacterial/probable bacterial meningitis (P<.001), compared with nonbacterial CNS infections and facial palsy, as well as with a control group without signs of CNS involvement. The values of CSF HGF were not correlated to blood-brain-barrier disruption in the groups. These observations might indicate an intrathecal production of HGF in acute bacterial/probable bacterial meningitis. PMID- 10837202 TI - Rifampin reduces production of reactive oxygen species of cerebrospinal fluid phagocytes and hippocampal neuronal apoptosis in experimental Streptococcus pneumoniae meningitis. AB - Bacterial compounds induce the production of reactive oxygen species (ROS) in meningitis. Rifampin releases smaller quantities of proinflammatory compounds from Streptococcus pneumoniae than do beta-lactam antibiotics. Therefore, rabbits infected intracisternally with S. pneumoniae were treated intravenously either with rifampin 5 mg/kg/h or ceftriaxone 10 mg/kg/h (n=9 each). Before initiation of antibiotic treatment, a strong positive correlation between ROS production of cerebrospinal fluid (CSF) phagocyte populations and bacterial CSF titers was observed (granulocytes: rs=.90, P<.0001; monocytes: rs=.81, P<.0001). CSF leukocytes from rifampin-treated rabbits produced less ROS (monocytes at 2 h after initiation of treatment: P=.045; at 5 h: P=.014; granulocytes at 5 h: P=.036) than did leukocytes from animals receiving ceftriaxone. The CSF malondialdehyde concentrations and the density of apoptotic neurons in the dentate gyrus were lower in rifampin- than in ceftriaxone-treated animals (P=.002 and.005). The use of rifampin to reduce the release of ROS and to decrease secondary brain injury appears promising. PMID- 10837203 TI - Vitamin D receptor gene polymorphisms and susceptibility to Mycobacterium malmoense pulmonary disease. AB - Polymerase chain reaction using sequence-specific primers was utilized to ascertain the prevalence of 3 polymorphisms of the vitamin D receptor (VDR) gene (FokI F/f, ApaI A/a, and TaqI T/t) in 56 patients with Mycobacterium malmoense pulmonary disease. When compared with 101 controls, M. malmoense patients displayed an increased prevalence of Apa1 A (P=.03; Fisher's exact test), TaqI t (P=.04), and the At VDR haplotype (P=.04), and they displayed a decreased prevalence of FokI f (P=.04). Only 4 (7%) of 56 patients (vs. 29 [28%] of 101 controls) were both positive for FokI f and negative for At (P=.001). This indicates that polymorphisms in the VDR (or in closely linked genes) modulate the susceptibility to M. malmoense and that susceptibility involves multiple genetic and environmental factors. PMID- 10837204 TI - Is there an association between Kawasaki disease and Chlamydia pneumoniae? AB - The clinical and epidemiological features of Kawasaki disease (KD) are consistent with an infectious cause. Because chronic infection with Chlamydia pneumoniae has been implicated in the pathogenesis of atherosclerosis, it has been suggested that it may also be involved in the pathogenesis of KD. Paired sera (baseline pretreatment and 1 year after treatment with intravenous immunoglobulin [IVIG]) from 26 children with KD and 29 age-matched controls were examined by microimmunofluorescence (MIF) serology and immunoblotting. There were no significant differences in the prevalence of anti-C. pneumoniae IgG, IgA, or IgM between cases and controls; however, 73%-85% of sera from cases and controls reacted with C. pneumoniae proteins by immunoblotting. There was significantly more reactivity in the pre-IVIG, but not post-IVIG, KD sera compared with sera from controls to proteins at 72-74 kDa and 74-76 kDa. They may be heat shock proteins. The results of this study do not support an association between KD and C. pneumoniae on the basis of MIF and immunoblot analysis. PMID- 10837205 TI - Genetic transformation of Coccidioides immitis facilitated by Agrobacterium tumefaciens. AB - Agrobacterium tumefaciens was used to facilitate genetic transformation of Coccidioides immitis. A gene cassette containing the gene encoding hygromycin phosphotransferase (hph) was cloned into a T-DNA vector plasmid and introduced into A. tumefaciens, and the resultant strain was used for cocultivation with germinated arthroconidia. This procedure produced numerous colonies 60- to >500 fold more resistant to hygromycin than untransformed mycelia. Both polymerase chain reaction and Southern blot analysis of the transformants indicated that all contained hph, usually as a single genomic copy. A transformation frequency of 1 per 10(5) arthroconidia was obtained by varying the germination time prior to cocultivation and altering the bacterium: fungus ratio. This approach requires no special equipment that might complicate biocontainment. Furthermore, transformation does not require digestion of fungal cell walls, further simplifying this procedure. A. tumefaciens-facilitated transformation should make possible the development of tagged mutagenesis and targeted gene disruption technology for C. immitis and perhaps other fungi of medical importance. PMID- 10837206 TI - Differential cellular accumulation of transforming growth factor-beta1, -beta2, and -beta3 in brains of patients who died with cerebral malaria. AB - In cerebral malaria (CM), pathologic cytokine expression patterns are thought to contribute to disruption of the blood-brain barrier, inflammation, and astrocytic scar formation. Expression of transforming growth factor (TGF)-beta1, -beta2, and -beta3 was analyzed in the brains of 7 patients who died with CM and in 8 control patients. In the brains of patients with CM, there were significantly (P=.0003) more TGF-beta1-immunoreactive astrocytes adjacent to brain vessels with deposition of malarial pigment, significantly (P=.0081) more TGF-beta2-expressing macrophages/microglial cells in glioses of ring hemorrhages and Durck's granulomas, and significantly (P=.0022) more TGF-beta3-expressing smooth-muscle cells and endothelial cells of brain vessels with sequestration. It is concluded that focal accumulation of TGF-beta1, -beta2, and -beta3 provides evidence for their involvement in the reorganization process of the brain parenchyma, immunologic dysfunction, and endothelial cell activation in patients with CM. PMID- 10837207 TI - Tracing Dobrava hantavirus infection. PMID- 10837208 TI - Haemophilus influenzae type B (Hib) antibody responses in children given diphtheria-tetanus-acellular pertussis-Hib combination vaccines. PMID- 10837209 TI - Reply. PMID- 10837210 TI - Role of Fas/FasL in systemic candidiasis. PMID- 10837211 TI - Reply. PMID- 10837212 TI - Immunology of reproduction: protecting the placenta. AB - The propensity of complement to damage self is controlled by expression of regulatory proteins. Recent results demonstrate that deleting just one of these regulators in mice causes complement to attack and destroy the embryo. These findings may have relevance to human pregnancy. PMID- 10837213 TI - Membrane lipids: it's only a phase. AB - All biological membranes contain lipids that prefer to adopt a non-bilayer phase. Recent results suggest that, in the thylakoid membrane, membrane proteins force all the lipids to adopt a bilayer structure, and that the non-bilayer-forming lipids in the thylakoid membrane serve to drive the formation of membrane stacks. PMID- 10837214 TI - Auditory neuroscience: is speech special? AB - Speech is thought to be perceived and processed in a unique way by the auditory system of the brain. A recent study has provided evidence that a part of the brain's temporal lobe is specifically responsive to speech and other vocal stimuli. PMID- 10837215 TI - Gene regulation: better late than never? AB - A new study has found that, in Arabidopsis, the paternal copies of many genes are delayed in expression during early seed development. The distribution of the genes and nature of their products suggest that this delayed expression of paternal alleles may be a global phenomenon. PMID- 10837216 TI - Neurotrophins: more to NGF than just survival. AB - NGF's role in neuronal survival has tended to mask other potential functions. But now the generation of mice lacking both Bax - required for the death of NGF deprived neurons - and NGF or its receptor has shown that NGF plays a key role in establishing cutaneous innervation and regulating neuropeptide expression. PMID- 10837217 TI - Insect mechanoreception: what a long, strange TRP it's been. AB - Insect bristles are model mechanosensory organs. An ion channel of the TRP superfamily has recently been identified which is required for production of mechanoreceptor currents by insect bristles, and seems likely to represent a new kind of mechanically gated channel. PMID- 10837218 TI - EGF receptor signalling: the importance of presentation. AB - Activation of the Drosophila EGF receptor requires the transmembrane TGF-alpha like ligand Spitz. Recent studies have shed new light on the role of two transmembrane proteins, Star and Rhomboid, in the presentation and subsequent proteolytic processing of Spitz. PMID- 10837219 TI - Ribosomal translocation: EF-G turns the crank. AB - Recent results from cryo-electron microscopy have shown that substantial structural rearrangements in both elongation factor EF-G and the ribosome occur during tRNA translocation. The observed sites of interaction between EF-G and the ribosome are consistent with molecular mimicry models for EF-G function. PMID- 10837220 TI - Drosophila dumpy is a gigantic extracellular protein required to maintain tension at epidermal-cuticle attachment sites. AB - BACKGROUND: Growth and morphogenesis during development depend both on patterning genes, which assign positional information, and on genes that regulate mechanical forces. The dumpy gene of the fruit fly Drosophila melanogaster is an example of the latter class, with mutant phenotypes affecting size and shape of the limbs, thoracic cuticle, trachea and mouthparts. RESULTS: The genetically complex dumpy locus was found to span over 100 kb and encode a gigantic 2.5 MDa extracellular matrix protein. Dumpy represents an extreme form of modular protein evolution, containing 308 epidermal growth factor (EGF) modules, interspersed with a new module class, DPY, and terminating in a crosslinking zona pellucida domain and membrane anchor sequence. We determined the three-dimensional structure of the DPY module by nuclear magnetic resonance (NMR) spectroscopy and found that it forms a disulphide-stabilised beta sheet motif, capable of linking end-to-end with EGF modules to form a fibre. Consistent with its cuticle phenotypes, dumpy is expressed at several sites of cuticle-epidermal cell attachment, including the trachea and the muscle tendon cells, which mediate anchorage of the muscles to the cuticle. CONCLUSIONS: The dumpy gene encodes a gigantic extracellular molecule that we predict to be a membrane-anchored fibre of almost a micrometer in length. Insertion and crosslinking of this fibre within the cuticle may provide a strong anchor for the underlying tissue, allowing it to maintain mechanical tension at sites under stress. This would explain its contribution to tissue morphogenesis through its regulation of mechanical properties. PMID- 10837221 TI - Regulation of the hTERT telomerase catalytic subunit by the c-Abl tyrosine kinase. AB - BACKGROUND: Telomeres consist of repetitive (TTAGGG) DNA sequences that are maintained by the multisubunit telomerase ribonucleoprotein. Telomerase consists of an RNA, which serves as template for the sequence tracts, and a catalytic subunit that functions in reverse transcription of the RNA template. Cloning and characterization of the human catalytic subunit of telomerase (hTERT) has supported a role in cell transformation. How telomerase activity is regulated, however, is largely unknown. RESULTS: We show here that hTERT associates directly with the c-Abl protein tyrosine kinase. We also found that c-Abl phosphorylates hTERT and inhibits hTERT activity. Moreover, our findings demonstrate that exposure of cells to ionizing radiation induces tyrosine phosphorylation of hTERT by a c-Abl-dependent mechanism. The functional significance of the c-Abl-hTERT interaction is supported by the demonstration that cells deficient in c-Abl show telomere lengthening. CONCLUSIONS: The ubiquitously expressed c-Abl tyrosine kinase is activated by DNA double-strand breaks. Our finding of telomere lengthening in c-Abl-deficient cells and the functional interactions between c Abl and hTERT support a role for c-Abl in the regulation of telomerase function. PMID- 10837222 TI - Visualizing muscle cell migration in situ. AB - BACKGROUND: Cell migration has been studied extensively by manipulating and observing cells bathed in putative chemotactic or chemokinetic agents on planar substrates. This environment differs from that in vivo and, consequently, the cells can behave abnormally. Embryo slices provide an optically accessible system for studying cellular navigation pathways during development. We extended this system to observe the migration of muscle precursors from the somite into the forelimb, their cellular morphology, and the localization of green fluorescent protein (GFP)-tagged adhesion-related molecules under normal and perturbed conditions. RESULTS: Muscle precursors initiated migration synchronously and migrated in broad, rather than highly defined, regions. Bursts of directed migration were followed by periods of meandering or extension and retraction of cell protrusions. Although paxillin did not localize to discernible intracellular structures, we found that alpha-actinin localized to linear, punctate structures, and the alpha5 integrin to some focal complexes and/or vesicle-like concentrations. Alterations in the expression of adhesion molecules inhibited migration. The muscle precursors migrating in situ formed unusually large, long lived protrusions that were polarized in the direction of migration. Unlike wild type Rac, a constitutively active Rac localized continuously around the cell surface and promoted random protrusive activity and migration. CONCLUSIONS: The observation of cellular migration and the dynamics of molecular organization at high temporal and spatial resolution in situ is feasible. Migration from the somite to the wing bud is discontinuous and not highly stereotyped. In situ, local activation of Rac appears to produce large protrusions, which in turn, leads to directed migration. Adhesion can also regulate migration. PMID- 10837223 TI - The molecular basis of melanism and mimicry in a swallowtail butterfly. AB - Melanism in Lepidoptera, either industrial or in mimicry, is one of the most commonly cited examples of natural selection [1] [2]. Despite extensive studies of the frequency and maintenance of melanic genes in insect populations [1] [2], there has been little work on the underlying molecular mechanisms. Nowhere is butterfly melanism more striking than in the Eastern Tiger Swallowtail (Papilio glaucus) of North America [3] [4] [5]. In this species, females can be either yellow (wild type) or black (melanic). The melanic form is a Batesian mimic of the distasteful Pipevine Swallowtail (Battus philenor), which is also black in overall color. Melanism in P. glaucus is controlled by a single Y-linked (female) black gene [6]. Melanic females, therefore, always have melanic daughters. Black melanin replaces the background yellow in melanic females. Here, we show that the key enzyme involved is N-beta-alanyl-dopamine-synthase (BAS), which shunts dopamine from the melanin pathway into the production of the yellow color pigment papiliochrome and also provides products for cuticle sclerotization. In melanic females, this enzyme is suppressed, leading to abnormal melanization of a formerly yellow area, and wing scale maturation is also delayed in the same area. This raises the possibility that either reduced BAS activity itself is preventing scale sclerotization (maturation) or, in contrast, that the delay in scale maturation precludes expression of BAS at the correct stage. Together, these data show how changes in expression of a single gene product could result in multiple wing color phenotypes. The implications for the genetic control of mimicry in other Lepidoptera are discussed. PMID- 10837224 TI - Maternal-specific footprints at putative CTCF sites in the H19 imprinting control region give evidence for insulator function. AB - Parent-of-origin-specific expression of the mouse insulin-like growth factor 2 (Igf2) gene and the closely linked H19 gene are regulated by an intervening 2 kb imprinting control region (ICR), which displays parentspecific differential DNA methylation [1] [2]. Four 21 bp repeats are embedded within the ICR and are conserved in the putative ICR of human and rat Igf2 and H19, suggesting that the repeats have a function [3] [4]. Here, we report that prominent DNA footprints were found in vivo on the unmethylated maternal ICR at all four 21 bp repeats, demonstrating the presence of protein binding. The methylated paternal ICR displayed no footprints. Significantly, the maternal-specific footprints were localized to putative binding sites for CTCF, a highly conserved zinc-finger DNA binding protein with multiple roles in gene regulation including that of chromatin insulator function [5] [6]. These results strongly suggest that the maternal ICR functions as an insulator element in regulating mutually exclusive expression of Igf2 and H19 in cis. PMID- 10837225 TI - The cyclin-dependent kinase Cdk5 controls multiple aspects of axon patterning in vivo. AB - Cyclin-dependent kinase 5 (Cdk5) is one of a subfamily of Cdks involved in the control of cell differentiation and morphology rather than cell division. Specifically, Cdk5 and its activating subunit, p35, have been implicated in growth cone motility during axon extension. Both Cdk5 and p35 are expressed in post-mitotic neurons and are localized to growth cones [1] [2] [3] [4]. The Cdk5 p35 complex interacts with the Rac GTPase, a protein required for growth cone motility [5]. Studies using cultured neurons have suggested that Cdk5 activity controls the efficiency of neurite extension [3] [4]. Mutant mice lacking p35 exhibit subtle axon-guidance defects [6], but these mice have severe defects in neuronal migration [6] [7] [8], making it difficult to define precisely the role of the Cdk5-p35 complex in vivo. Here, we examined Cdk5 function in axon patterning in the Drosophila embryo. Although our data support the idea that Cdk5 p35 is involved in axonogenesis, they do not support the view that Cdk5 simply promotes growth cone motility. Instead, we found that disrupting Cdk5 function caused widespread errors in axon patterning. PMID- 10837226 TI - Optokinetic behavior is reversed in achiasmatic mutant zebrafish larvae. AB - The vertebrate optokinetic nystagmus (OKN) is a compensatory oculomotor behavior that is evoked by movement of the visual environment. It functions to stabilize visual images on the retina. The OKN can be experimentally evoked by rotating a drum fitted with stripes around the animal and has been studied extensively in many vertebrate species, including teleosts. This simple behavior has earlier been used to screen for mutations affecting visual system development in the vertebrate model organism zebrafish. In such a screen, we have found a significant number of homozygous belladonna (bel) mutant larvae to be defective in the correct execution of the OKN [1]. We now show that about 40% of homozygous bel larvae display a curious reversal of the OKN upon visual stimulation. Monocular stimulation leads to primary activation of ipsilateral eye movements in larvae that behave like the wild type. In contrast, affected larvae display contralateral activation of eye movements upon monocular stimulation. Anatomical analysis of retinal ganglion cell axon projections reveal a morphological basis for the observed behavioral defect. All animals with OKN reversal are achiasmatic. Further behavioral examination of affected larvae show that OKN reversed animals execute this behavior in a stimulus-velocity-independent manner. Our data support a parsimonious model of optokinetic reversal by the opening of a controlling feedback loop at the level of the optic chiasm that is solely responsible for the observed behavioral abnormality in mutant belladonna larvae. PMID- 10837227 TI - Simulated self-motion alters perceived time to collision. AB - Many authors have assumed that motor actions required for collision avoidance and for collision achievement (for example, in driving a car or hitting a ball) are guided by monitoring the time to collision (TTC), and that this is done on the basis of moment-to-moment values of the optical variable tau [1] [2] [3]. This assumption has also motivated the search for single neurons that fire when tau is a certain value [4] [5] [6] [7] [8]. Almost all of the laboratory studies and all the animal experiments were restricted to the case of stationary observer and moving object. On the face of it, this would seem reasonable. Even though humans and other animals routinely perform visually guided actions that require the TTC of an approaching object to be estimated while the observer is moving, tau provides an accurate estimate of TTC regardless of whether the approach is produced by self-motion, object-motion or a combination of both. One might therefore expect that judgements of TTC would be independent of self-motion. We report here, however, that simulated selfmotion using a peripheral flow field substantially altered estimates of TTC for an approaching object, even though the peripheral flow field did not affect the value of tau for the approaching object. This finding points to long range interactions between collision-sensitive visual neurons and neural mechanisms for processing self-motion. PMID- 10837228 TI - The role of pre- and post-anaphase microtubules in the cytokinesis phase of the cell cycle. AB - The cytokinesis phase, or C phase, of the cell cycle results in the separation of one cell into two daughter cells after the completion of mitosis. Although it is known that microtubules are required for proper positioning of the cytokinetic furrow [1] [2], the role of pre-anaphase microtubules in cytokinesis has not been clearly defined for three key reasons. First, inducing microtubule depolymerization or stabilization before the onset of anaphase blocks entry into anaphase and cytokinesis via the spindle checkpoint [3]. Second, microtubule organization changes rapidly at anaphase onset as the mitotic kinase, Cdc2-cyclin B, is inactivated [4]. Third, the time between the onset of anaphase and the initiation of cytokinesis is very short, making it difficult to unambiguously alter microtubule polymer levels before cytokinesis, but after inactivation of the spindle checkpoint. Here, we have taken advantage of the discovery that microinjection of antibodies to the spindle checkpoint protein Mad2 (mitotic arrest deficient) in prometaphase abrogates the spindle checkpoint, producing premature chromosome separation, segregation, and normal cytokinesis [5] [6]. To test the role of pre-anaphase microtubules in cytokinesis, microtubules were disassembled in prophase and prometaphase cells, the cells were then injected with anti-Mad2 antibodies and recorded through C phase. The results show that exit from mitosis in the absence of microtubules triggered a 50 minute period of cortical contractility that was independent of microtubules. Furthermore, upon microtubule reassembly during this contractile C-phase period, approximately 30% of the cells underwent chromosome poleward movement, formed a midzone microtubule complex, and completed cytokinesis. PMID- 10837229 TI - Dynamics of cytoplasmic dynein in living cells and the effect of a mutation in the dynactin complex actin-related protein Arp1. AB - Cytoplasmic dynein is a minus-end-directed microtubule motor that participates in multiple cellular activities such as organelle transport and mitotic spindle assembly [1]. To study the dynamic behavior of cytoplasmic dynein in the filamentous fungus Aspergillus nidulans, we replaced the gene for the cytoplasmic dynein heavy chain, nudA, with a gene encoding a green fluorescent protein (GFP) tagged chimera, GFP-nudA. The GFP-NUDA fusion protein is fully functional in vivo: strains expressing only the GFP-tagged nudA grow as well as wild-type strains. Fluorescence microscopy showed GFP-NUDA to be in comet-like structures that moved in the hyphae toward the growing tip. Retrograde movement of some GFP NUDA comets after they arrived at the tip was also observed. These dynamics of GFP-NUDA were not observed in cells treated with a microtubule-destabilizing drug, benomyl, suggesting they are microtubule-dependent. The rate of GFP-NUDA tip-ward movement is similar to the rate of cytoplasmic microtubule polymerization toward the hyphal tip, suggesting that GFP-NUDA is associated and moving with the polymerizing ends of microtubules. A mutation in actin-related protein Arp1 of the dynactin complex abolishes the presence of these dynamic GFP NUDA structures near the hyphal tip, suggesting a targeting role of the dynactin complex. PMID- 10837231 TI - Mob1p interacts with the Sid2p kinase and is required for cytokinesis in fission yeast. AB - A great deal is now known about how cells regulate entry into mitosis, but only recently have the mechanisms controlling exit from mitosis and cytokinesis begun to be revealed. In the budding yeast Saccharomyces cerevisiae, Mob1p interacts with the Dbf2p kinase and cells containing mutations in these genes arrest in late anaphase [1] [2]. Proteins related to Mob1p are present in both plants and animals, but information about Mob1p function has been obtained only from budding yeast. Here, we describe the identification and characterization of Mob1p from Schizosaccharomyces pombe. Mob1p associates with the Sid2p kinase and like Sid2p, Mob1p is required for the initiation of cytokinesis, but not for mitotic exit. Mob1p localizes to the spindle pole body (SPB) and to the cell-division site during cell division, suggesting that it might be involved in transducing the signal to initiate cell division from the SPB to the division site. Mob1p is required for Sid2p localization, and Mob1p localization requires the function of the cdc7, cdc11, cdc14, spg1, sid1, sid2, and sid4 genes, suggesting that together with Sid2p, Mob1p functions at the end of the signaling cascade required to regulate the onset of cytokinesis at the end of mitosis. PMID- 10837230 TI - Cdc14 activates cdc15 to promote mitotic exit in budding yeast. AB - Inactivation of mitotic cyclin-dependent kinases (Cdks) is required for cells to exit mitosis [1] [2]. In the budding yeast Saccharomyces cerevisiae, Cdk inactivation is triggered by the phosphatase Cdc14, which is activated by a complex network of regulatory proteins that includes the protein kinase Cdc15 [3] [4] [5] [6]. Here we show that the ability of Cdc15 to promote mitotic exit is inhibited by phosphorylation. Cdc15 is phosphorylated in vivo at multiple Cdk consensus sites during most of the cell cycle, but is transiently dephosphorylated in late mitosis. Although phosphorylation appears to have no effect on Cdc15 kinase activity, a non-phosphorylatable mutant of Cdc15 is a more potent stimulator of mitotic exit than wild-type Cdc15, indicating that phosphorylation inhibits Cdc15 function in vivo. Interestingly, inhibitory phosphorylation of Cdc15 is removed by the phosphatase Cdc14 in vitro, and overproduction of Cdc14 leads to Cdc15 dephosphorylation in vivo. Thus, Cdc15 serves both as an activator and substrate of Cdc14. Although this scheme raises the possibility that positive feedback promotes Cdc14 activation, we present evidence that such feedback is not essential for Cdc14 activation in vivo. Instead, Cdc15 dephosphorylation may promote some additional function of Cdc15 that is independent of its effects on Cdc14 activation. PMID- 10837233 TI - Inside a rat's brain PMID- 10837232 TI - Statistically significant sites PMID- 10837234 TI - A natural selection PMID- 10837235 TI - Homologous recombination. PMID- 10837236 TI - Protein kinase B. PMID- 10837237 TI - Skin development: delta laid bare. AB - Notch signalling is best known for its role in lateral inhibition, where it acts to prevent differentiation of cells neighbouring one that has 'won out' in a competition to differentiate. Recent results suggest that Notch signalling can work in the opposite way, and promote differentiation of the receiving cells. PMID- 10837238 TI - From sensory evidence to a motor command. AB - New insight into how the brain makes a decision has come from a study of the effects of the decision-making process on an eye movement evoked by electrical stimulation of the frontal cortex. The accumulation of sensory evidence was found to cause a gradual commitment toward a choice. PMID- 10837239 TI - Mammalian development: new trick for an old dog. AB - A recent study has shown that the T-box gene eomesodermin, which was first identified in Xenopus, not only plays a conserved role in vertebrates directing gastrulation, but interestingly in mammals has acquired a new function in the development of the trophoblast. PMID- 10837240 TI - Eye-hand coordination: eye to hand or hand to eye? AB - Single-unit recording has revealed both hand and eye movement-related activity in the parietal cortex of the macaque monkey. These experiments, as well as neuropsychological studies, are unravelling the complex nature of how the eye and the hand work together in the control of visually guided movements. PMID- 10837241 TI - What determines the rate of sequence evolution? AB - High rates of amino-acid sequence evolution have sometimes been considered to be diagnostic for genes undergoing adaptive change. However, two recent studies have shown that rapid evolution of amino-acid sequence can also be congruent with neutrality. PMID- 10837242 TI - Membrane transport: tethers and TRAPPs. AB - Transport vesicles are tethered to their target membrane prior to the interaction of v-SNAREs and t-SNAREs across the membrane junction. Recent evidence suggests tethering is a complex process requiring multiple components. PMID- 10837243 TI - Signal transduction: response regulators on and off. AB - High resolution structures of the active phosphorylated forms of two-component response regulators have recently been reported. The results provide a basis for understanding how metabolic energy is coupled to signal transduction in cellular regulatory networks. PMID- 10837244 TI - Protein sorting: recognizing mitochondrial presequences. AB - The mitochondrial protein import machinery specifically recognizes many different preproteins lacking a consensus sequence. The three-dimensional structure of an import receptor complexed to an amino-terminal targeting 'presequence' provides exciting insight into the molecular mechanism of signal recognition. PMID- 10837245 TI - Phosphorylation of the MEKK Ste11p by the PAK-like kinase Ste20p is required for MAP kinase signaling in vivo. AB - BACKGROUND: Many signals are transduced from the cell surface to the nucleus through mitogen-activated protein (MAP) kinase cascades. Activation of MAP kinase requires phosphorylation by MEK, which in turn is controlled by Raf, Mos or a group of structurally related kinases termed MEKKs. It is not understood how MEKKs are regulated by extracellular signals. In yeast, the MEKK Ste11p functions in multiple MAP kinase cascades activated in response to pheromones, high osmolarity and nutrient starvation. Genetic evidence suggests that the p21 activated protein kinase (PAK) Ste20p functions upstream of Ste11p, and Ste20p has been shown to phosphorylate Ste11p in vitro. RESULTS: Ste20p phosphorylated Ste11p on Ser302 and/or Ser306 and Thr307 in yeast, residues that are conserved in MEKKs of other organisms. Mutating these sites to non-phosphorylatable residues abolished Ste11p function, whereas changing them to aspartic acid to mimic the phosphorylated form constitutively activated Ste11p in vivo in a Ste20p independent manner. The amino-terminal regulatory domain of Ste11p interacted with its catalytic domain, and overexpression of a small amino-terminal fragment of Ste11p was able to inhibit signaling in response to pheromones. Mutational analysis suggested that this interaction was regulated by phosphorylation and dependent on Thr596, which is located in the substrate cleft of the catalytic domain. CONCLUSIONS: Our results suggest that, in response to multiple extracellular signals, phosphorylation of Ste11p by Ste20p removes an amino terminal inhibitory domain, leading to activation of the Ste11 protein kinase. This mechanism may serve as a paradigm for the activation of mammalian MEKKs. PMID- 10837246 TI - Evidence from functional magnetic resonance imaging of crossmodal binding in the human heteromodal cortex. AB - BACKGROUND: Integrating information from the different senses markedly enhances the detection and identification of external stimuli. Compared with unimodal inputs, semantically and/or spatially congruent multisensory cues speed discrimination and improve reaction times. Discordant inputs have the opposite effect, reducing performance and slowing responses. These behavioural features of crossmodal processing appear to have parallels in the response properties of multisensory cells in the superior colliculi and cerebral cortex of non-human mammals. Although spatially concordant multisensory inputs can produce a dramatic, often multiplicative, increase in cellular activity, spatially disparate cues tend to induce a profound response depression. RESULTS: Using functional magnetic resonance imaging (fMRI), we investigated whether similar indices of crossmodal integration are detectable in human cerebral cortex, and for the synthesis of complex inputs relating to stimulus identity. Ten human subjects were exposed to varying epochs of semantically congruent and incongruent audio-visual speech and to each modality in isolation. Brain activations to matched and mismatched audio-visual inputs were contrasted with the combined response to both unimodal conditions. This strategy identified an area of heteromodal cortex in the left superior temporal sulcus that exhibited significant supra-additive response enhancement to matched audio-visual inputs and a corresponding sub-additive response to mismatched inputs. CONCLUSIONS: The data provide fMRI evidence of crossmodal binding by convergence in the human heteromodal cortex. They further suggest that response enhancement and depression may be a general property of multisensory integration operating at different levels of the neuroaxis and irrespective of the purpose for which sensory inputs are combined. PMID- 10837247 TI - The caspase-8 inhibitor FLIP promotes activation of NF-kappaB and Erk signaling pathways. AB - BACKGROUND: Activation of Fas (CD95) by its ligand (FasL) rapidly induces cell death through recruitment and activation of caspase-8 via the adaptor protein Fas associated death domain protein (FADD). However, Fas signals do not always result in apoptosis but can also trigger a pathway that leads to proliferation. We investigated the level at which the two conflicting Fas signals diverge and the protein(s) that are implicated in switching the response. RESULTS: Under conditions in which proliferation of CD3-activated human T lymphocytes is increased by recombinant FasL, there was activation of the transcription factors NF-kappaB and AP-1 and recruitment of the caspase-8 inhibitor and FADD interacting protein FLIP (FLICE-like inhibitory protein). Fas-recruited FLIP interacts with TNF-receptor associated factors 1 and 2, as well as with the kinases RIP and Raf-1, resulting in the activation of the NF-kappaB and extracellular signal regulated kinase (Erk) signaling pathways. In T cells these two signal pathways are critical for interleukin-2 production. Increased expression of FLIP in T cells resulted in increased production of interleukin-2. CONCLUSIONS: We provide evidence that FLIP is not simply an inhibitor of death receptor-induced apoptosis but that it also mediates the activation of NF-kappaB and Erk by virtue of its capacity to recruit adaptor proteins involved in these signaling pathways. PMID- 10837248 TI - Tribbles, a cell-cycle brake that coordinates proliferation and morphogenesis during Drosophila gastrulation. AB - BACKGROUND: The final shape and size of an organism is determined by both morphogenetic processes and cell proliferation and it is essential that these processes be properly coordinated. In particular, cell division is incompatible with certain types of morphogenetic cell behaviour, such as migration, adhesion and changes in cell shape. Mechanisms must therefore exist to ensure that one does not interfere with the other. RESULTS: We address here the coordination of proliferation and morphogenesis during the development of the mesoderm in Drosophila. We show that it is essential that mitosis be blocked in the mesoderm during early gastrulation, and identify the putative serine/threonine kinase Tribbles as controlling this block. In its absence, the mitotic block is lifted, resulting in severe defects during early gastrulation. Tribbles, a homologue of a group of vertebrate proteins of unknown function, acts in concert with another, as yet unidentified, factor to counteract the activity of the protein phosphatase Cdc25/String. CONCLUSIONS: In a finely tuned balance with Cdc25/String, Tribbles controls the timing of mitosis in the prospective mesoderm, allowing cell-shape changes to be completed. This mechanism for coordinating cell division and cell shape changes may have helped Drosophila to evolve its mode of rapid early development. PMID- 10837249 TI - Genetic identification of neurons controlling a sexually dimorphic behaviour. AB - In the fruit fly Drosophila melanogaster, locomotor activity is sexually dimorphic: female flies constantly modulate their activity pattern whereas males show a steadier, stereotyped walking pace [1]. Here, we mapped the area of the brain controlling this behavioural dimorphism. Adult male Drosophila expressing a dominant feminising transgene in a small cluster of neurons in the pars intercerebralis exhibited a female-like pattern of locomotor activity. Genetic ablation of these neurons prevented the feminisation of the locomotor activity of transgenic males. The results suggest that this cluster of neurons modulates sex specific activity, but is not involved in initiating fly locomotion. Nor does it control male courtship behaviour, because feminisation of courtship was not correlated with the feminisation of locomotor activity. PMID- 10837250 TI - Laminin A is required for follicle cell-oocyte signaling that leads to establishment of the anterior-posterior axis in Drosophila. AB - The establishment of the anterior-posterior (AP) axis in Drosophila melanogaster requires signaling between the oocyte and surrounding somatic follicle cells during oogenesis [1] [2]. First, a signal from the oocyte (Gurken (Grk), a transforming growth factor-alpha (TGFalpha) homolog) is received by predetermined terminal follicle cells in which the epidermal growth factor receptor (EGFR) pathway is activated and a posterior fate is induced [2] [3] [4]. Later, the posterior follicle cells send an unidentified signal back to the oocyte, which leads to the reorganization of its cytoskeletal polarity. This reorganization is required for proper localization of maternal determinants, such as oskar (osk) and bicoid (bcd) mRNAs, that determine the AP polarity of the oocyte and the subsequent embryo [2]. We show here that when the gene lanA, which encodes the extracellular matrix component laminin A, is mutated in posterior follicle cells, localization of AP determinants is disrupted in the underlying oocyte. Posterior follicle-cell differentiation and follicle cell apical-basal polarity are unaffected in the lanA mutant cells, suggesting that laminin A is required for correct signaling from the posterior follicle cells that polarizes the oocyte. This is the first evidence that the extracellular matrix is involved in the establishment of a major body axis. PMID- 10837251 TI - Cyclic AMP regulates the HERG K(+) channel by dual pathways. AB - Lethal cardiac arrhythmias are a hallmark of the hereditary Long QT syndrome (LQTS), a disease produced by mutations of cardiac ion channels [1]. Often these arrhythmias are stress-induced, suggesting a relationship between beta-adrenergic activation of adenylate cyclase and cAMP-dependent alteration of one or more of the ion channels involved in LQTS. Second messengers modulate ion channel activity either by direct interaction or through intermediary kinases and phosphatases. Here we show that the second messenger cAMP regulates the K(+) channel mutated in the LQT2 form of LQTS, HERG [2], both directly and indirectly. Activation of cAMP-dependent protein kinase (PKA) causes phosphorylation of HERG accompanied by a rapid reduction in current amplitude, acceleration of voltage dependent deactivation, and depolarizing shift in voltage-dependent activation. In a parallel pathway, cAMP directly binds to the HERG protein with the opposing effect of a hyperpolarizing shift in voltage-dependent activation. The summation of cAMP-mediated effects is a net diminution of the effective current, but when HERG is complexed with with the K(+) channel accessory proteins MiRP1 or minK, the stimulatory effects of cAMP are favored. These findings provide a direct link between stress and arrhythmia by a unique mechanism where a single second messenger exerts complex regulation of an ion channel via two distinct pathways. PMID- 10837252 TI - Deriving behavioural receptive fields for visually completed contours. AB - The visual system is constantly faced with the problem of identifying partially occluded objects from incomplete images cast on the retinae. Phenomenologically, the visual system seems to fill in missing information by interpolating illusory and occluded contours at points of occlusion, so that we perceive complete objects. Previous behavioural [1] [2] [3] [4] [5] [6] [7] and physiological [8] [9] [10] [11] [12] studies suggest that the visual system treats illusory and occluded contours like luminance-defined contours in many respects. None of these studies has, however, directly shown that illusory and occluded contours are actually used to perform perceptual tasks. Here, we use a response-classification technique [13] [14] [15] [16] [17] [18] [19] [20] to answer this question directly. This technique provides pictorial representations - 'classification images' - that show which parts of a stimulus observers use to make perceptual decisions, effectively deriving behavioural receptive fields. Here we show that illusory and occluded contours appear in observers' classification images, providing the first direct evidence that observers use perceptually interpolated contours to recognize objects. These results offer a compelling demonstration of how visual processing acts on completed representations, and illustrate a powerful new technique for constraining models of visual completion. PMID- 10837253 TI - Coherent global motion in the absence of coherent velocity signals. AB - It is widely believed that form and motion are analysed separately in mammalian visual systems. Form is confined within a stream that projects ventrally from V1 to the inferotemporal cortex, and motion within a stream that projects more dorsally, to the posterior parietal cortex [1] [2] [3] [4] [5] [6] [7]. Current descriptions suggest that there is little contact between the two streams until the products of their separate analyses are bound together at a late (and still unidentified) stage in perception [3] [8] [9] [10]. There are, however, indications that form and motion signals may interact [11], and that form signals, streaks derived from motion, may assist in the analysis of its direction [12]. Lennie [13] proposes that all image attributes, form and motion included, remain intimately coupled within the same retinotopic map at all stages of visual analysis. Here we show that form, independent of motion, can give coherence to incoherent motion. Sequences of Glass patterns [14] built to a common global rule are devoid of coherent motion signals, but they produce motion consistent with the global rule for form, not with the random velocity components of the pattern sequence. PMID- 10837254 TI - A mutation in the Lunatic fringe gene suppresses the effects of a Jagged2 mutation on inner hair cell development in the cochlea. AB - Recent studies have demonstrated that the Notch signaling pathway regulates the differentiation of sensory hair cells in the vertebrate inner ear [1] [2] [3] [4] [5] [6] [7] [8] [9]. We have shown previously that in mice homozygous for a targeted null mutation of the Jagged2 (Jag2) gene, which encodes a Notch ligand, supernumerary hair cells differentiate in the cochlea of the inner ear [7]. Other components of the Notch pathway, including the Lunatic fringe (Lfng) gene, are also expressed during differentiation of the inner ear in mice [6] [7] [8] [9] [10]. In contrast to the Jag2 gene, which is expressed in hair cells, the Lfng gene is expressed in non-sensory supporting cells in the mouse cochlea [10]. Here we demonstrate that a mutation in the Lfng gene partially suppresses the effects of the Jag2 mutation on hair cell development. In mice homozygous for targeted mutations of both Jag2 and Lfng, the generation of supernumerary hair cells in the inner hair cell row is suppressed, while supernumerary hair cells in the outer hair cell rows are unaffected. We also demonstrate that supernumerary hair cells are generated in mice heterozygous for a Notch1 mutation. We suggest a model for the action of the Notch signaling pathway in regulating hair cell differentiation in the cochlear sensory epithelium. PMID- 10837255 TI - Complex formation between Mad1p, Bub1p and Bub3p is crucial for spindle checkpoint function. AB - The spindle checkpoint delays the metaphase to anaphase transition in response to defects in kinetochore-microtubule interactions in the mitotic apparatus (see [1] [2] [3] [4] for reviews). The Mad and Bub proteins were identified as key components of the spindle checkpoint through budding yeast genetics [5] [6] and are highly conserved [3]. Most of the spindle checkpoint proteins have been localised to kinetochores, yet almost nothing is known about the molecular events which take place there. Mad1p forms a tight complex with Mad2p [7], and has been shown to recruit Mad2p to kinetochores [8]. Similarly, Bub3p binds to Bub1p [9] and may target it to kinetochores [10]. Here, we show that budding yeast Mad1p has a regulated association with Bub1p and Bub3p during a normal cell cycle and that this complex is found at significantly higher levels once the spindle checkpoint is activated. We find that formation of this complex requires Mad2p and Mps1p but not Mad3p or Bub2p. In addition, we identify a conserved motif within Mad1p that is essential for Mad1p-Bub1p-Bub3p complex formation. Mutation of this motif abolishes checkpoint function, indicating that formation of the Mad1p-Bub1p-Bub3p complex is a crucial step in the spindle checkpoint mechanism. PMID- 10837256 TI - Mitsuhiro Yanagida: cutting apart mitosis. PMID- 10837257 TI - The 14-3-3 proteins. PMID- 10837259 TI - Crystal clear PMID- 10837258 TI - Genome sequencing: mind the gap. PMID- 10837260 TI - The future in our hands. PMID- 10837261 TI - Ca(2+) signalling and control of guard-cell volume in stomatal movements. AB - Stomatal guard cells are unique as a plant cell model and, because of the depth of knowledge now to hand on ion transport and its regulation, serve as an excellent model for the analysis of stimulus-response coupling in higher plants. Parallel controls - mediated by Ca(2+), H(+) protein kinases and phosphatases - regulate the gating of the K(+) and Cl(-) channels that facilitate solute flux for stomatal movements. A growing body of evidence now indicates that oscillations in the cytosolic free concentration of Ca(2+) contribute to a 'signalling cassette', which is integrated within these events through an unusual coupling with membrane voltage. Additional developments during the past two years point to events in membrane traffic that play complementary roles in stomatal control. Research in these areas, especially, is now adding entirely new dimensions to our understanding of guard cell signalling. PMID- 10837262 TI - Phytochelatin biosynthesis and function in heavy-metal detoxification. AB - Plants respond to heavy-metal toxicity via a number of mechanisms. One such mechanism involves the chelation of heavy metals by a family of peptide ligands, the phytochelatins. Molecular genetic approaches have resulted in important advances in our understanding of phytochelatin biosynthesis. In particular, genes encoding the enzyme phytochelatin synthase have been isolated from plant and yeast species. Unexpectedly, genes with similar sequences to those encoding phytochelatin synthase have been identified in some animal species. PMID- 10837263 TI - Ascorbic acid: metabolism and functions of a multi-facetted molecule. AB - Ascorbic acid (vitamin C) is the most abundant antioxidant in plants. Its biosynthetic pathway via GDP-D-mannose and L-galactose, which was proposed only recently, is now supported by molecular genetic evidence from Arabidopsis thaliana and transgenic potato plants. Except for the last step (which is located on the inner mitochondrial membrane) the pathway is cytosolic, sharing GDP-sugar intermediates with cell-wall polysaccharide and glycoprotein synthesis. Ascorbate peroxidase is emerging as a key enzyme in the fine control of H(2)O(2) concentration; its expression being controlled by redox signals and H(2)O(2). Convincing evidence of the involvement of ascorbate in cell division and growth is also accumulating. Its role as a cofactor in the synthesis of cell wall hydroxyproline-rich glycoproteins is one mechanism for this function. PMID- 10837264 TI - Unraveling plant metabolism by EST analysis. AB - Large-scale single-pass sequencing of cDNAs prepared from specific plant species or tissues has evolved as an inexpensive and efficient gene-discovery tool that can be used to identify novel cDNAs encoding enzymes of specific plant metabolic pathways. Collections of expressed sequence tags from metabolically active tissues can provide quantitative estimates of gene expression levels and thus are being exploited to unravel plant metabolic and regulatory networks. PMID- 10837265 TI - Molecular responses to dehydration and low temperature: differences and cross talk between two stress signaling pathways. AB - Recently, a major transcription system that controls abscisic-acid-independent gene expression in response to dehydration and low temperature has been identified. The system includes the DRE/CRT (dehydration-responsive element/C repeat) cis-acting element and its DNA-binding protein, DREB/CBF (DRE-binding protein/C-repeat binding factor), which has an AP2 domain. DREB/CBF contains two subclasses, DREB1/CBF and DREB2, which are induced by cold and dehydration, respectively, and control the expression of various genes involved in stress tolerance. Recent studies are providing evidence of differences between dehydration-signaling and cold-stress-signaling cascades, and of cross-talk between them. PMID- 10837266 TI - Mathematical modelling of metabolism. AB - Modelling of metabolism attempts to improve our understanding of metabolic regulation by quantifying essential parts or aspects of the metabolic system. Three areas in which modelling has recently made considerable contributions toward this aim can be identified. First, the more detailed description of individual reactions and pathways; second, the analysis of relative flux limitations within a pathway by means of metabolic control analysis; and third, in vivo flux analysis using nuclear magnetic resonance or mass spectroscopic analysis in combination with positionally labelled carbon compounds. PMID- 10837267 TI - The molecular physiology of ammonium uptake and retrieval. AB - Plants are able to take up ammonium from the soil, or through symbiotic interactions with microorganisms, via the root system. Using functional complementation of yeast mutants, it has been possible to identify a new class of membrane proteins, the ammonium transporter/methylammonium permease (AMT/MEP) family, that mediate secondary active ammonium uptake in eukaryotic and prokaryotic organisms. In plants, the AMT gene family can be subdivided according to their amino-acid sequences into three subfamilies: a large subfamily of AMT1 genes and two additional subfamilies each with single members (LeAMT1;3 from tomato and AtAMT2;1 from Arabidopsis thaliana). These transporters vary especially in their kinetic properties and regulatory mechanism. High-affinity transporters are induced in nitrogen-starved roots, whereas other transporters may be considered as the 'work horses' that are active when conditions are conducive to ammonium assimilation. The expression of several AMTs in root hairs further supports a role in nutrient acquisition. These studies provide basic information that will be needed for the dissection of nitrogen uptake by plants at the molecular level and for determining the role of individual AMTs in nutrient uptake and potentially in nutrient efficiency. PMID- 10837268 TI - Concerted hydrogen-atom abstraction in photosynthetic water oxidation. AB - Photosystem II evolves oxygen by using water in the unlikely role of a reductant. The absorption of sunlight by chlorophyll produces highly oxidizing equivalents that are filled with electrons stripped from water. This proton-coupled redox chemistry occurs at the oxygen-evolving complex, which contains a tetramanganese cluster, a redox-active tyrosine amino acid hydrogen-bonded to a histidine amino acid, a calcium ion and chloride. Hydrogen-atom abstraction by the tyrosyl radical from water bound to the manganese cluster is now widely held to occur in this process, at least for some of the steps in the catalytic cycle. We discuss kinetic and energetic constraints on the hydrogen-atom abstraction process. PMID- 10837269 TI - Exploring the possibilities presented by protein engineering. AB - A combination of classical and powerful new combinatorial genetic techniques allows the redesign of enzyme activities and creation of proteins that are tailored to have specific properties. These technologies have far-reaching consequences for the future design of crop plants and the storage compounds within them. PMID- 10837270 TI - Regulation of sulfate transport and synthesis of sulfur-containing amino acids. AB - Recent research indicates that several sulfate transporters - exhibiting different tissue specificities and modes of expression - may play distinct roles in sulfate uptake within specific tissues and in long-distance sulfate translocation. The transcription levels of particular genes and feedback inhibition of serine acetyltransferase play major roles in regulating sulfur assimilation and cysteine synthesis. O-acetylserine and glutathione presumably act within the cysteine synthesis pathway as derepressor and repressor, respectively. A unique autoregulatory mechanism that stabilizes mRNA levels has recently been proposed for the regulation of methionine synthesis. PMID- 10837271 TI - Delivering copper within plant cells. AB - Two genes recently identified in Arabidopsis thaliana may be involved in sequestering free copper ions in the cytoplasm and delivering copper to post Golgi vesicles. The genes COPPER CHAPERONE and RESPONSIVE TO ANTAGONIST1 are homologous to copper-trafficking genes from yeast and humans. This plant copper delivery pathway is required to create functional ethylene receptors. The pathway may also facilitate the transport of copper from senescing leaf tissue. In addition, several other genes have been identified recently that may have a role in copper salvage during senescence. PMID- 10837272 TI - Phosphate transport and signaling. AB - The discovery of phosphate (Pi) transporter genes has provided a basis for the molecular study of the complex pattern of Pi transport in plants. Over the past two years, a significant amount of information has been generated on the molecular regulation of phosphate transport in plants. Recent developments in plant genomics will soon allow the complete dissection of the signal transduction pathway(s) associated with plant responses to Pi limitation in the rhizosphere. PMID- 10837273 TI - Calcium requirement is a sliding scale. PMID- 10837274 TI - The important role of physical activity in skeletal development: how exercise may counter low calcium intake. PMID- 10837275 TI - Implications of racial distinctions for body composition and its diagnostic assessment. PMID- 10837276 TI - Genetic underpinnings of LDL size and density: a role for hepatic lipase? PMID- 10837277 TI - Measures of body composition in blacks and whites: a comparative review. AB - Biological differences exist in the body composition of blacks and whites. We reviewed literature on the differences and similarities between the 2 races relative to fat-free body mass (water, mineral, and protein), fat patterning, and body dimensions and proportions. In general, blacks have a greater bone mineral density and body protein content than do whites, resulting in a greater fat-free body density. Additionally, there are racial differences in the distribution of subcutaneous fat and the length of the limbs relative to the trunk. The possibility that these differences are a result of ethnicity rather than of race is also examined. Because most equations that predict relative body fat were derived from predominantly white samples, biological variation between the races in these body-composition indexes has practical significance. Systematic error can result in the inaccurate estimation of the relative body fat of blacks, and therefore of definitions of obesity, if these inherent differences are ignored. PMID- 10837278 TI - Virtual seminars for disseminating medical nutrition education curriculum ideas. AB - There is a need and a desire for educators working toward implementation of nutrition in medical schools and residency programs to share ideas and materials. The World Wide Web enables computer-mediated communications through which a medical nutrition curriculum could be discussed; however, existing formats lack focus and structure. In January 1999, a virtual seminar that focused on nutrition education in medical schools and residency programs was conducted. The seminar, titled "Making Room for Nutrition Education, was sponsored by organizations that have active medical nutrition educators. The seminar included 5 topics discussed over a 4-d period. The transcript was made available at http://www.preventivenutrition. com. There were 119 registered participants. Responses to a postseminar questionnaire were positive; there was interest in an ongoing series of virtual seminars. PMID- 10837280 TI - Metabolic assessment of female chronic dieters with either normal or low resting energy expenditures. AB - BACKGROUND: Chronic dieting syndrome can have negative physiologic and psychological consequences. Metabolic differences between female chronic dieters with normal and with low resting energy expenditures (REEs) have not been fully examined. OBJECTIVE: To determine whether differences existed between 2 groups (n = 15/group) of female chronic dieters aged 21-49 y with either normal (>/=100% of predicted) and with low (/=9 micromol/L participated in a randomized, controlled trial to compare 3 approaches for increasing dietary folate to approximately 600 microg/d: folic acid supplementation, consumption of folic acid-fortified breakfast cereals, and increased consumption of folate-rich foods. RESULTS: An intake of 437 microg folic acid/d from supplements resulted in a 27-nmol/L increase in serum folate and a 21% reduction in tHcy, relative to the change in a control group. In subjects who consumed folic acid-fortified breakfast cereal, folate intake increased by an average of 298 microg, serum folate increased by 21 nmol/L, and tHcy concentrations decreased by 24%. Increased intakes of folate-rich foods resulted in a 418-microg increase in dietary folate, a 7-nmol/L increase in serum folate, and a 9% reduction in tHcy concentrations. The decrease in tHcy was negatively correlated (r = -0.66) with the increase in serum folate. CONCLUSIONS: Daily consumption of folic acid fortified breakfast cereals and the use of folic acid supplements appear to be the most effective means of reducing tHcy concentrations. The reduction in tHcy was significantly negatively correlated with the increase in serum folate, which may be a useful marker for measuring dietary change. PMID- 10837285 TI - A prospective study of dietary glycemic load, carbohydrate intake, and risk of coronary heart disease in US women. AB - BACKGROUND: Little is known about the effects of the amount and type of carbohydrates on risk of coronary heart disease (CHD). OBJECTIVE: The objective of this study was to prospectively evaluate the relations of the amount and type of carbohydrates with risk of CHD. DESIGN: A cohort of 75521 women aged 38-63 y with no previous diagnosis of diabetes mellitus, myocardial infarction, angina, stroke, or other cardiovascular diseases in 1984 was followed for 10 y. Each participant's dietary glycemic load was calculated as a function of glycemic index, carbohydrate content, and frequency of intake of individual foods reported on a validated food-frequency questionnaire at baseline. All dietary variables were updated in 1986 and 1990. RESULTS: During 10 y of follow-up (729472 person years), 761 cases of CHD (208 fatal and 553 nonfatal) were documented. Dietary glycemic load was directly associated with risk of CHD after adjustment for age, smoking status, total energy intake, and other coronary disease risk factors. The relative risks from the lowest to highest quintiles of glycemic load were 1.00, 1.01, 1. 25, 1.51, and 1.98 (95% CI: 1.41, 2.77 for the highest quintile; P for trend < 0.0001). Carbohydrate classified by glycemic index, as opposed to its traditional classification as either simple or complex, was a better predictor of CHD risk. The association between dietary glycemic load and CHD risk was most evident among women with body weights above average ?ie, body mass index (in kg/m(2)) >/= 23. CONCLUSION: These epidemiologic data suggest that a high dietary glycemic load from refined carbohydrates increases the risk of CHD, independent of known coronary disease risk factors. PMID- 10837286 TI - Soy isoflavones improve plasma lipids in normocholesterolemic, premenopausal women. AB - BACKGROUND: Soy consumption is known to reduce plasma total cholesterol and LDL cholesterol in hypercholesterolemic subjects, but the responsible soy components and the effects in normocholesterolemic subjects remain unclear. OBJECTIVE: The effects of soy isoflavone consumption on plasma total cholesterol, HDL cholesterol, LDL-cholesterol, triacylglycerol, apolipoprotein A-I, apolipoprotein B, and lipoprotein(a) concentrations and on LDL peak particle diameter were examined in normocholesterolemic, premenopausal women. DESIGN: Thirteen healthy, normocholesterolemic, free-living, premenopausal female volunteers took part in this randomized, crossover-controlled trial. Each subject acted as her own control. Three soy isoflavone intakes (control: 10.0 +/- 1.1; low: 64.7 +/- 9.4; and high: 128.7 +/- 15.7 mg/d), provided as soy protein isolate, were consumed for 3 menstrual cycles each. Total cholesterol, HDL cholesterol, LDL cholesterol, and triacylglycerol were measured over the menstrual cycle. Apolipoprotein A-I, apolipoprotein B, lipoprotein(a), and LDL peak particle diameter were evaluated in the midluteal phase. RESULTS: Total cholesterol, HDL-cholesterol, and LDL cholesterol concentrations changed significantly across menstrual cycle phases (P < 0.005). During specific phases of the cycle, the high-isoflavone diet lowered LDL cholesterol by 7.6-10.0% (P < 0.05), the ratio of total cholesterol to HDL cholesterol by 10.2% (P < 0.005), and the ratio of LDL to HDL cholesterol by 13.8% (P < 0.002). CONCLUSIONS: Isoflavones significantly improved the lipid profile across the menstrual cycle in normocholesterolemic, premenopausal women. Although of small magnitude, these effects could contribute to a lower risk of developing coronary heart disease in healthy people who consume soy over many years. PMID- 10837287 TI - Uptake of individual fatty acids into adipose tissue in relation to their presence in the diet. AB - BACKGROUND: The fatty acid composition of adipose tissue triacylglycerol reflects, but is not identical to, the fatty acid composition of the habitual diet. OBJECTIVE: We investigated whether the fatty acid composition of adipose tissue is explained by differences between fatty acids in early storage in adipose tissue after a meal. DESIGN: Nine healthy men ate a meal containing several fatty acids. Blood samples were taken for 6 h after the meal from an arterialized hand vein and a vein draining the anterior abdominal subcutaneous adipose tissue. RESULTS: Net storage of fatty acids in adipose tissue occurred between 1 and 4 h after the meal. In relation to the amount fed, storage of fatty acids differed (P < 0. 01) between classes (n-3 polyunsaturated < saturated < n-6 polyunsaturated < monounsaturated); oleic acid was stored in the greatest amounts. These differences agreed closely with published data, except for n-3 polyunsaturated fatty acids. The only individual metabolic step at which significant differences between fatty acids was shown was incorporation of fatty acids into chylomicron triacylglycerol. Differences between fatty acids in rate of extraction from chylomicron triacylglycerol and net uptake into adipose tissue in the postprandial period were significant (P < 0. 01), but not when expressed in relation to proportions in chylomicron triacylglycerol. CONCLUSIONS: The characteristic fatty acid pattern of adipose tissue may predominantly reflect the early metabolic handling of different fatty acids. Adipose tissue uptake of n-3 polyunsaturated fatty acids is slow in relation to that of other fatty acids. PMID- 10837288 TI - Temporal trends in energy intake in the United States: an ecologic perspective. AB - BACKGROUND: The causes of recent increases in the prevalence of overweight in the United States are perplexing because national survey data do not show clear patterns of change in energy intake or expenditure. OBJECTIVE: Ecologic data regarding the quantities and types of foods and nutrients available in the United States between 1970 and 1998 were reviewed to provide an alternative perspective on trends in energy intake. DESIGN: Literature searches in agriculture, business, and medical library databases were conducted to identify data regarding the quantities and types of foods and nutrients available in the United States between 1970 and 1998. RESULTS: Per capita energy availability estimates from the US Department of Agriculture, US Food Supply Series, indicate that energy availability increased by 15% between 1970 and 1994. Data regarding trends in food purchasing and preparation suggest that Americans are eating more meals outside the home, relying more heavily on convenience foods, and consuming larger food portions. Americans appear, in some cases, to be selecting lower-fat foods over higher-fat alternatives. In addition, availability and sales of reduced energy and reduced-fat products have increased. CONCLUSION: Consistent with trends in overweight, most of the ecologic data reviewed suggest that energy intake has increased over the past several decades and is likely a major contributor to increases in average body weight. PMID- 10837289 TI - Lack of hemoglobin response to iron supplementation in anemic mexican preschoolers with multiple micronutrient deficiencies. AB - BACKGROUND: In developing countries, incomplete resolution of anemia with iron supplementation is often attributed to poor compliance or inadequate duration of supplementation, but it could result from deficiencies of other micronutrients. OBJECTIVE: Our objective was to assess children's hematologic response to supervised, long-term iron supplementation and the relation of this response to other micronutrient deficiencies, anthropometry, morbidity, and usual dietary intake. DESIGN: Rural Mexican children aged 18-36 mo (n = 219) were supplemented for 12 mo with either 20 mg Fe, 20 mg Zn, both iron and zinc, or placebo. Children were categorized as iron-unsupplemented (IUS; n = 109) or iron supplemented (IS; n = 108). Hemoglobin, hematocrit, mean corpuscular volume, mean cell hemoglobin, plasma concentrations of micronutrients that can affect hematopoiesis, anthropometry, and diet were assessed at 0, 6, and 12 mo; morbidity was assessed biweekly. RESULTS: At baseline, 70% of children had low hemoglobin (40 nmol/L, 1,25(OH)(2)D was positively related to 25(OH)D; in women with 25(OH)D concentrations 40 nmol/L, 1,25(OH)(2)D was most closely (inversely) related to plasma creatinine. Therefore, with serum 25(OH)D concentrations increasingly <40 nmol/L, serum 1,25(OH)(2)D becomes critically dependent on rising concentrations of PTH. CONCLUSION: The data suggest that aging women should maintain 25(OH)D concentrations >40 nmol/L (which is the lower limit of our normal range for healthy young subjects) for optimal bone health. PMID- 10837303 TI - Human milk oligosaccharides are resistant to enzymatic hydrolysis in the upper gastrointestinal tract. AB - BACKGROUND: Human milk oligosaccharides (HMOs) show a complexity and variety not found in milk of any other species. Although progress has been made in the past 3 decades with regard to identification and structural characterization of HMOs, not much is known about the physiologic functions of HMOs. OBJECTIVE: As a prerequisite for biological activity in infant metabolism, HMOs have to resist enzymatic hydrolysis in the gastrointestinal tract. To assess the extent to which selected HMOs are hydrolyzed, we carried out in vitro digestion studies using enzyme preparations of human and porcine pancreas and intestinal brush border membranes (BBMs). DESIGN: Fractions of HMOs, including structurally defined isolated oligosaccharides, were digested for up to 20 h with human pancreatic juice and BBMs prepared from human or porcine intestinal tissue samples. HMOs were incubated by using a porcine pancreatic homogenate and BBMs as enzyme sources. HMOs and digestion products were identified by mass spectrometry and anion-exchange chromatography. Additionally, free D-glucose, L-fucose, and N acetylneuraminic acid were determined enzymatically. RESULTS: Whereas maltodextrin (control) was rapidly and completely hydrolyzed, neutral and acidic HMOs showed a profound resistance against pancreatic juice and BBM hydrolases. However, cleavage of most of the HMOs was achieved by using a pancreatic homogenate containing intracellular, including lysosomal, enzymes in addition to secreted enzymes. CONCLUSIONS: The results of this study strongly suggest that HMOs are not hydrolyzed by enzymes in the upper small intestine. Although intact HMOs may be absorbed, we postulate that a majority of HMOs reach the large intestine, where they serve as substrates for bacterial metabolism. Therefore, HMOs might be considered the soluble fiber fraction of human milk. PMID- 10837302 TI - Relation of serum retinol to acute phase proteins and malarial morbidity in Papua New Guinea children. AB - BACKGROUND: Acute phase proteins (APPs) are associated with malaria-induced hyporetinemia (serum retinol <0.70 micromol/L); however, the degree of the association is not well documented. OBJECTIVE: The association between malaria induced hyporetinemia and APPs was assessed. DESIGN: In a cross-sectional study, 90 children with serum retinol concentrations from <0.35 to >1.05 micromol/L were selected from children in a clinical trial of vitamin A supplementation. Serum was collected before treatment allocation. Retinol binding protein (RBP) concentrations were determined by radioimmunoassays, and transthyretin, alpha(1) acid glycoprotein (AGP), alpha(1)-antichymotrypsin, C-reactive protein (CRP), haptoglobin, and albumin concentrations by radial immunodiffusion assays. RESULTS: Children in the subsample had high rates of splenomegaly and Plasmodium positive blood-smear slides (P < 0.01); AGP (Pearson's r = -0.40, P < 0.001) and CRP (r = -0.21, P = 0.04) were inversely correlated with retinol. The negative APPs RBP, transthyretin, and albumin were positively and significantly associated with retinol. All APPs, except alpha(1)-antichymotrypsin, were significantly correlated with splenomegaly. Of the positive APPs, AGP correlated with CRP (r = 0.37, P < 0.001), indicating chronic inflammation. In a stepwise regression analysis, 73% of retinol's variability was explained by RBP and transthyretin. The model predicted that a 1-SD increase in RBP or transthyretin increases retinol by approximately 0.38 or 0.47 micromol/L, respectively, whereas an equivalent increase in AGP decreases retinol by 0.12 micromol/L. CONCLUSIONS: The RBP-transthyretin transport complex of retinol is not altered by inflammation. Positive APPs are useful markers of type and severity of inflammation; however, except for AGP, it is unlikely that they can correct for malaria-induced hyporetinemia. PMID- 10837304 TI - Iron bioavailability in infants from an infant cereal fortified with ferric pyrophosphate or ferrous fumarate. AB - BACKGROUND: Infant cereals are commonly fortified with insoluble iron compounds with low relative bioavailability, such as ferric pyrophosphate, because of organoleptic changes that occur after addition of water-soluble iron sources. OBJECTIVE: Our objective was to compare iron bioavailability from ferric pyrophosphate with an alternative iron source that is soluble in dilute acid, ferrous fumarate, and to evaluate the influence of ascorbic acid on iron bioavailability from ferrous fumarate in infants. DESIGN: Iron bioavailability was measured as the incorporation of stable iron isotopes into erythrocytes 14 d after administration of labeled test meals (25 g dry wheat and soy infant cereal, 100 g water, and 2.5 mg Fe as [57Fe]ferric pyrophosphate or [57Fe]ferrous fumarate). Ascorbic acid was added to all test meals (25 mg in study 1 or 25 or 50 mg in study 2). Infants were fed each test meal on 4 consecutive days under standardized conditions. The 2 different test meals within each study were administered 2 wk apart in a crossover design. RESULTS: Geometric mean iron bioavailability was significantly higher from [57Fe]ferrous fumarate than from [57Fe]ferric pyrophosphate [4.1% (range: 1.7-14.7%) compared with 1.3% (range: 0. 7-2.7%); n = 8, P = 0.008]. In this study, doubling the ascorbic acid content did not further enhance iron bioavailability; the geometric means (range) were 3.4% (1.9-6.6%) and 4.2% (1.2-18.7%) for the test meals with 25 and 50 mg ascorbic acid added, respectively (n = 9). CONCLUSION: Iron bioavailability from iron fortified infant cereals can be improved by using an iron compound with high relative bioavailability and by ensuring adequate ascorbic acid content of the product. PMID- 10837305 TI - Minimal enteral nutrient requirements for intestinal growth in neonatal piglets: how much is enough? AB - BACKGROUND: Parenterally nourished preterm infants commonly receive minimal enteral feedings, the aim being to enhance intestinal function. Whether this regimen increases intestinal growth has not been established. OBJECTIVE: Our objective was to determine the minimal enteral nutrient intakes necessary to stimulate and to normalize neonatal intestinal growth. METHODS: Intestinal growth and cell proliferation were quantified in neonatal pigs given equal amounts of an elemental nutrient solution for 7 d. Different groups (n = 5-7 per group) received 0%, 10%, 20%, 40%, 60%, 80%, or 100% of total nutrient intake enterally, with the remainder given parenterally. RESULTS: In the jejunum, wet weight, protein mass, and villus height were significantly greater at enteral intakes >40%. Stimulation of ileal protein mass required a higher enteral intake (60%). In both segments, abrupt increases in DNA mass, crypt depth, ornithine decarboxylase activity, and crypt cells in S-phase occurred between enteral intakes of 40% and 60%. Circulating concentrations of glucagon-like peptide-2 and peptide YY, but not gastrin, increased significantly between enteral intakes of 40% and 60% and closely paralleled indexes of cell proliferation. CONCLUSIONS: The minimal enteral nutrient intake necessary to increase mucosal mass was 40% of total nutrient intake, whereas 60% enteral nutrition was necessary to sustain normal mucosal proliferation and growth. Our results imply that providing <40% of the total nutrient intake enterally does not have significant intestinal trophic effects. PMID- 10837308 TI - Variability of k terms in bioelectrical impedance spectroscopy. PMID- 10837306 TI - Reduced LDL particle size in children consuming a very-low-fat diet is related to parental LDL-subclass patterns. AB - BACKGROUND: A genetically influenced atherogenic lipoprotein phenotype characterized by a predominance of small, dense LDL particles (subclass pattern B) can be induced by low-fat diets in healthy subjects with large LDL particles (pattern A). OBJECTIVE: The aim of this study was to test whether genetic predisposition to subclass pattern B contributes to susceptibility to induction of this trait by a low-fat diet. DESIGN: The prevalence of pattern B in children is relatively low compared with that in older individuals, but genetic susceptibility to this trait in offspring can be inferred by its presence in their parents. Plasma lipoproteins were analyzed 10 d after a change from a usual diet to a very-low-fat (10% fat), high-carbohydrate diet in offspring (mean age: 14 y; range: 7-28 y) of 22 families according to parental LDL-subclass patterns when consuming a low-fat diet: AxA mating (9 families with 19 children), AxB mating (5 families with 10 children), and BxB mating (8 families with 21 children). RESULTS: The very-low-fat, high-carbohydrate diet produced significantly greater decreases in LDL particle size in offspring of BxB parents (f1.gif" BORDER="0"> +/- SE: -0.55 +/- 0.16 nm) and AxB parents (-0.48 +/- 0.19 nm) than in offspring of AxA parents (0.14 +/- 0.20 nm). The number of children expressing pattern B with the 10%-fat diet and the proportion of children converting from pattern A to pattern B was significantly greater in offspring of BxB parents than in those with 1 or 2 pattern A parents. CONCLUSION: A very-low fat, high-carbohydrate diet can induce expression of LDL-subclass pattern B in genetically predisposed children with low expression of the trait while consuming their usual diets. PMID- 10837309 TI - Introduction and conference goals PMID- 10837310 TI - Functional foods: the US perspective. AB - Widespread interest in the possibility that selected foods might promote health has resulted in the coining of the term functional food, although agreement about what is and what is not a functional food is lacking. Public interest in functional foods is increasing because of higher health care costs; the passage of federal legislation affecting many food categories, including the expanded category of dietary supplements; and recent scientific discoveries linking dietary habits with the development of many diseases, including coronary heart disease and some cancers. A variety of foods have been proposed as providing health benefits by altering one or more physiologic processes. Biomarkers are needed to assess the ability of functional foods or their bioactive components to modify disease and to evaluate the ability of these foods to promote health, growth, and well-being. Evidence suggests that several biomarkers may be useful for distinguishing between diseased and nondiseased states and even for predicting future susceptibility to disease. A variety of biomarkers will probably be needed to develop a profile for an individual that reflects the impact of diet on performance and health. Another area of interest is the interaction of nutrients and their association with genetics. These interactions may account for the inconsistent interrelations observed between specific dietary constituents and the incidence of disease. Greater understanding of how diet influences a person's genetic potential, overall performance, and susceptibility to disease can have enormous implications for society. As new discoveries are made in this area, consumers will need access to this information so that they can make informed decisions. PMID- 10837311 TI - Concepts and strategy of functional food science: the European perspective. AB - Recent knowledge supports the hypothesis that, beyond meeting nutrition needs, diet may modulate various functions in the body and play detrimental or beneficial roles in some diseases. Concepts in nutrition are expanding from emphasis on survival, hunger satisfaction, and preventing adverse effects to emphasizing the use of foods to promote a state of well-being and better health and to help reduce the risk of disease. In many countries, especially Japan and the United States, research on functional foods is addressing the physiologic effects and health benefits of foods and food components, with the aim of authorizing specific health claims. The positive effects of a functional food can be either maintaining a state of well-being and health or reducing the risk of pathologic consequences. Among the most promising targets for functional food science are gastrointestinal functions, redox and antioxidant systems, and metabolism of macronutrients. Ongoing research into functional foods will allow the establishment of health claims that can be translated into messages for consumers that will refer to either enhanced function or reduction of disease risk. Only a rigorous scientific approach that produces highly significant results will guarantee the success of this new discipline of nutrition. This presents a challenge for the scientific community, health authorities, and the food industry. PMID- 10837312 TI - Effect of functional food ingredients: vitamin E modulation of cardiovascular diseases and immune status in the elderly. AB - Increased accumulation of free radicals over time reduces the effectiveness of antioxidant defense mechanisms and heightens the vulnerability of older individuals to a variety of oxidative insults and associated pathologic conditions. Both nutritive and nonnutritive components of foods may slow declines in certain body functions. Ingestion of vitamin E, an antioxidant nutrient, in amounts above current recommendations may reduce the risk of cardiovascular disease, enhance immune status, and otherwise modulate important degenerative conditions associated with aging. Early adoption of proper dietary habits helps adults to maintain quality of life as they age. Increased intake of vitamin E through selection of foods with large amounts of that vitamin and daily consumption of 5-8 servings of fruit and vegetables may reduce risk for cardiovascular disease and improve immune function in later life. PMID- 10837313 TI - Can nutrient supplements modify brain function? AB - Over the past 40 y, several lines of investigation have shown that the chemistry and function of both the developing and the mature brain are influenced by diet. Examples are the effect of folate deficiency on neural tube development during early gestation, the influence of essential fatty acid deficiency during gestation and postnatal life on the development of visual function in infants, and the effects of tryptophan or tyrosine intake (alone or as a constituent of dietary protein) on the production of the brain neurotransmitters derived from them (serotonin and the catecholamines, respectively). Sometimes the functional effects are clear and the underlying biochemical mechanisms are not (as with folate and essential fatty acids); in other cases (such as the amino acids tyrosine and tryptophan), the biochemical effects are well understood, whereas the effect on brain function is not. Despite the incomplete knowledge base on the effects of such nutrients, investigators, physicians, and regulatory bodies have promoted the use of these nutrients in the treatment of disease. Typically, these nutrients have been given in doses above those believed to be required for normal health; after they have been given in pure form, unanticipated adverse effects have occasionally occurred. If this pharmacologic practice is to continue, it is important from a public safety standpoint that each nutrient be examined for potential toxicities so that appropriate purity standards can be developed and the risks weighed against the benefits when considering their use. PMID- 10837315 TI - Nutritionally induced oxidative stress: effect on viral disease. AB - It has long been known that the nutritional status of the host can influence both susceptibility to infectious disease and the severity of the disease if contracted. In studies of coxsackievirus infection and selenium deficiency in mice, we found that mice fed a selenium-deficient diet developed myocarditis, but mice fed a diet adequate in selenium did not. Similarly, mice fed a diet deficient in vitamin E developed myocarditis, but mice fed a diet with adequate vitamin E did not. The epidemic of optic and peripheral neuropathy that occurred in Cuba in the early 1990s provides another example of how the nutritional status of the host may affect the impact of a virus. Patients who developed neuropathy had lower blood concentrations of riboflavin, vitamin E, selenium, alpha- and beta-carotenes, and the carotenoid lycopene, which suggests that the disease was associated with an impairment of protective antioxidant pathways. After supplementation of the population with these nutrients, the disease began to subside. The nutritional status of the host can have a profound influence on a virus, so that a normally avirulent virus becomes virulent because of changes in the viral genome. Our studies suggest that outbreaks of disease attributed to a nutritional deficiency may actually result from infection by a virus that has become pathogenic by replicating in a nutritionally deficient host. PMID- 10837317 TI - Prebiotics and probiotics: are they functional foods? AB - A probiotic is a viable microbial dietary supplement that beneficially affects the host through its effects in the intestinal tract. Probiotics are widely used to prepare fermented dairy products such as yogurt or freeze-dried cultures. In the future, they may also be found in fermented vegetables and meats. Several health-related effects associated with the intake of probiotics, including alleviation of lactose intolerance and immune enhancement, have been reported in human studies. Some evidence suggests a role for probiotics in reducing the risk of rotavirus-induced diarrhea and colon cancer. Prebiotics are nondigestible food ingredients that benefit the host by selectively stimulating the growth or activity of one or a limited number of bacteria in the colon. Work with prebiotics has been limited, and only studies involving the inulin-type fructans have generated sufficient data for thorough evaluation regarding their possible use as functional food ingredients. At present, claims about reduction of disease risk are only tentative and further research is needed. Among the claims are constipation relief, suppression of diarrhea, and reduction of the risks of osteoporosis, atherosclerotic cardiovascular disease associated with dyslipidemia and insulin resistance, obesity, and possibly type 2 diabetes. The combination of probiotics and prebiotics in a synbiotic has not been studied. This combination might improve the survival of the bacteria crossing the upper part of the gastrointestinal tract, thereby enhancing their effects in the large bowel. In addition, their effects might be additive or even synergistic. PMID- 10837319 TI - Lycopene and cardiovascular disease. AB - Considerable evidence suggests that lycopene, a carotenoid without provitamin A activity found in high concentrations in a small set of plant foods, has significant antioxidant potential in vitro and may play a role in preventing prostate cancer and cardiovascular disease in humans. Tomato products, including ketchup, tomato juice, and pizza sauce, are the richest sources of lycopene in the US diet, accounting for >80% of the total lycopene intake of Americans. Unlike other carotenoids, lycopene is not consistently lower among smokers than among nonsmokers, suggesting that any possible preventive activity is not as an antioxidant. Instead, lycopene may have a cholesterol synthesis-inhibiting effect and may enhance LDL degradation. Available evidence suggests that intimal wall thickness and risk of myocardial infarction are reduced in persons with higher adipose tissue concentrations of lycopene. The question of whether lycopene helps to prevent cardiovascular disease can only be answered by a trial specifically evaluating its effectiveness in this area. PMID- 10837321 TI - Tea polyphenols: prevention of cancer and optimizing health. AB - The tea plant Camellia sinesis is cultivated in >30 countries. Epidemiologic observations and laboratory studies have indicated that polyphenolic compounds present in tea may reduce the risk of a variety of illnesses, including cancer and coronary heart disease. Most studies involved green tea, however; only a few evaluated black tea. Results from studies in rats, mice, and hamsters showed that tea consumption protects against lung, forestomach, esophagus, duodenum, pancreas, liver, breast, colon, and skin cancers induced by chemical carcinogens. Other studies showed the preventive effect of green tea consumption against atherosclerosis and coronary heart disease, high blood cholesterol concentrations, and high blood pressure. Because the epidemiologic studies and research findings in laboratory animals have shown the chemopreventive potential of tea polyphenols in cancer, the usefulness of tea polyphenols for humans should be evaluated in clinical trials. One such phase 1 clinical trial is currently under way at the MD Anderson Cancer Center in collaboration with Memorial Sloan Kettering Cancer Center. This study will examine the safety and possible efficacy of consuming the equivalent of > or =10 cups (> or =2.4 L) of green tea per day. The usefulness of tea polyphenols may be extended by combining them with other consumer products such as food items and vitamin supplements. This "designer item" approach may be useful for human populations, but it requires further study. PMID- 10837323 TI - Protection against breast cancer with genistein: a component of soy. AB - Breast cancer is the most common cancer in women. Because genetics is believed to account for only 10-15% of breast cancer cases, the environment, including nutrition, is thought to play a significant role in predisposing women to this cancer. Studies of Asian women suggest that those who consume a traditional diet high in soy products have a low incidence of breast cancer, but that among emigrants to the United States, the second generation, but not the first, loses this protection. These findings suggest a possible common mechanism of action for breast cancer protection from early, specific nutritional exposure. Genistein, an isoflavone found in soy, has been reported to have weak estrogenic and antiestrogenic properties, to be an antioxidant, to inhibit topoisomerase II and angiogenesis, and to induce cell differentiation. In studies of the mammary glands of immature rats, we showed that genistein up-regulates the expression of the epidermal growth factor receptor shortly after treatment, which may be responsible for the increased cell proliferation seen at that age. We hypothesize that the early genistein action promotes cell differentiation that results in a less active epidermal growth factor signaling pathway in adulthood that, in turn, suppresses the development of mammary cancer. We speculate that breast cancer protection in Asian women consuming a traditional soy-containing diet is derived from early exposure to soybean products containing genistein. We believe that early events are essential for the benefits of cancer protection. PMID- 10837325 TI - Approaches for chronic disease prevention based on current understanding of underlying mechanisms. AB - Much progress has been achieved by exploring the causes of the main human cancers and of cardiovascular and cerebrovascular diseases. Even more important has been the knowledge acquired about the mechanisms underlying the development of these diseases. In many parts of the world, particularly in the West, the major cancers associated with dietary habits involve the postmenopausal breast, distal colon, prostate, pancreas, ovary, and endometrium. Current evidence suggests that the genotoxic carcinogens for all but the last 2 of these diseases stem from the traditional intake of fried and broiled foods such as meats. The surface of these foods contains a class of powerful mutagens, heterocyclic amines, which are carcinogenic to the target organs in animal models. Fish-eating populations have lower incidences of heart disease and of many types of cancers than do other populations, which may be the result of the n-3 polyunsaturated oils found in fish. Among other dietary practices that may reduce the risk of cancer and cardiovascular disease are consuming 5-9 servings of fruits and vegetables daily, which provides antioxidants such as quercetin and isothiocyanates; having a high fiber intake, including bran cereal; and drinking 1.5-2.5 L of fluids daily. Tea polyphenols found in black and green tea may have a protective effect against heart disease and some cancers. Concentrates of such desirable products have been made available in pill form to complement health-promoting personal lifestyles. Biomedical research funded by The National Institutes of Health and organizations such as the American Cancer Society has produced sound results that could lead to prevention of chronic disease. The public must heed this information to achieve long-term health. PMID- 10837328 TI - Use of T cell function to determine the effect of physiologically active food components. AB - The interdependency between the disciplines of nutrition and immunology was recognized in the 1970s when immunologic measures were introduced as a component of assessing nutritional status. Today, the immune response is considered integral to the pathophysiology of many chronic diseases in which diet plays a major role in prevention or treatment. T lymphocytes are an important adaptive cellular component of the immune system. Because of the difficulty in quantifying and isolating T cell function through clinical measures and in vivo immune challenges, most assessments of the effect of nutrition on immunity have been performed in vitro. A frequently used in vitro method to assess the cell-mediated response to nutritional intervention is lymphocyte blastogenesis. During the past 20 y, many soluble factors (cytokines) that influence cells involved in the immune and inflammatory responses have been described. Changes in dietary fat can modulate cytokine production in the absence of disease. Apoptosis (programmed cell death) is an exciting new area; a decrease in the rate of apoptosis may play a role in the pathogenesis of autoimmune disease and age-related events such as tumorigenesis. Energy restriction increases apoptosis. The goal of studying biomarkers of immune function is to understand how specific nutrients or foods directly and indirectly affect immunity. Biomarkers must be identified that can predict with reasonable accuracy resistance to infection and other illnesses associated with poor immune function. PMID- 10837330 TI - Functional foods: an ecologic perspective. AB - A functional food is defined as any food or food ingredient that may provide a health benefit beyond that conferred by the nutrients the food contains. As nutrition scientists move into this arena, they must build on the wealth of information that already exists in plant biology. In particular, the evolutionary and physiologic bases for the production of secondary plant chemicals in plants must be considered in order to plan meaningful experiments for testing the functionality of these chemical compounds for humans. One problem that may arise is that in using the term functional food, the meaning may be lost in the continued proliferation of related terms used in product marketing. The new National Institutes of Health Office of Dietary Supplements addressed some of these issues as it developed the operating definitions described in this report. PMID- 10837331 TI - Functional foods: the Food and Drug Administration perspective. AB - Because the Federal Food, Drug, and Cosmetic Act (FFDCA) does not provide a statutory definition of functional foods, the Food and Drug Administration has no authority to establish a formal regulatory category for such foods. The primary determinant of the regulatory status of a food is its intended use, which is determined largely by the label and labeling information accompanying the product. This information includes nutrient information, nutrient content claims, and various types of health claims. In marketing these foods, manufacturers may come under one of several existing regulatory options. The first decision manufacturers will make that will help determine their product's regulatory status is whether the product is a food or a drug. Thus, manufacturers and retailers have a range of legal and regulatory categories in which their products may be classified. This article describes the definitions provided in the FFDCA for a drug and a food, the safety and labeling requirements of various food categories, and types of possible claims for dietary supplements. PMID- 10837333 TI - Moderated discussion PMID- 10837334 TI - Summary PMID- 10837335 TI - Properties of cytotoxic peptide-formed ion channels. AB - Cytotoxic peptides are relatively small cationic molecules such as those found 1) in venoms, e.g., melittin in bee, scorpion toxins in scorpion, pilosulin 1 in jumper ant, and lycotoxin I and II in wolf spider; 2) in skin secretions (e.g., magainin I and II from Xenopus laevis, dermaseptin from frog, antimicrobials from carp) and cells of the immune system (e.g., insect, scorpion, and mammalian defensins and cryptdins); 3) as autocytotoxicity peptides, e.g., amylin cytotoxic to pancreatic beta-cells, prion peptide fragment 106-126 [PrP-(106-126)], and amyloid beta-protein (AbetaP) cytotoxic to neurons; and 4) as designed synthetic peptides based on the sequences and properties of naturally occurring cytotoxic peptides. The small cytotoxic peptides are composed of beta-sheets, e.g., mammalian defensins, AbetaP, amylin, and PrP-(106-126), whereas the larger cytotoxic peptides have several domains composed of both alpha-helices and beta sheets stabilized by cysteine bonds, e.g., scorpion toxins, scorpion, and insect defensins. Electrophysiological and molecular biology techniques indicate that these structures modify cell membranes via 1) interaction with intrinsic ion transport proteins and/or 2) formation of ion channels. These two nonexclusive mechanisms of action lead to changes in second messenger systems that further augment the abnormal electrical activity and distortion of the signal transduction causing cell death. PMID- 10837336 TI - Viscosity and solute dependence of F-actin translocation by rabbit skeletal heavy meromyosin. AB - We tested the hypothesis that solvent viscosity affects translocation of rhodamine phalloidin-labeled F-actin by rabbit skeletal heavy meromyosin (HMM). When viscosity was increased using either glycerol, fructose, sucrose, or dextran (1.5, 6.0, or 15-20 kDa mol mass), there was little or no effect on the fraction of moving filaments, whereas sliding speed decreased in inverse proportion to viscosity. The results could be explained neither by an effect of osmotic pressure at high solute concentrations nor by altered solvent drag on the actin filament. Elevated viscosity inhibited HMM ATPase activity in solution, but only at much higher viscosities than were needed to reduce sliding speed. Polyethylene glycols (300, 1,000, or 3,000 mol wt) also inhibited speed via elevated viscosity but secondarily inhibited by enhancing electrostatic interactions. These results demonstrate that a diffusion-controlled process intrinsic to cross-bridge cycling can be limiting to actomyosin function. PMID- 10837337 TI - Two distinct mechanisms of nitric oxide-mediated neuronal cell death show thiol dependency. AB - To better understand the mechanism(s) underlying nitric oxide (. NO)-mediated toxicity, in the presence and absence of concomitant oxidant exposure, postmitotic terminally differentiated NT2N cells, which are incapable of producing. NO, were exposed to PAPA-NONOate (PAPA/NO) and 3-morpholinosydnonimine (SIN-1). Exposure to SIN-1, which generated peroxynitrite in the range of 25-750 nM/min, produced a concentration- and time-dependent delayed cell death. In contrast, a critical threshold concentration (>440 nM/min) was required for. NO to produce significant cell injury. Examination of cells by electron microscopy shows a largely necrotic injury after peroxynitrite exposure but mainly apoptotic like morphology after. NO exposure. Cellular levels of reduced thiols correlated with cell death, and pretreatment with N-acetylcysteine (NAC) fully protected from cell death in either PAPA/NO or SIN-1 exposure. NAC given within the first 3 h posttreatment further delayed cell death and increased the intracellular thiol level in SIN-1 but not. NO-exposed cells. Cell injury from. NO was independent of cGMP, caspases, and superoxide or peroxynitrite formation. Overall, exposure of non-. NO-producing cells to. NO or peroxynitrite results in delayed cell death, which, although occurring by different mechanisms, appears to be mediated by the loss of intracellular redox balance. PMID- 10837338 TI - Skeletal muscle function and water permeability in aquaporin-4 deficient mice. AB - It has been proposed that aquaporin-4 (AQP4), a water channel expressed at the plasmalemma of skeletal muscle cells, is important in normal muscle physiology and in the pathophysiology of Duchenne's muscular dystrophy. To test this hypothesis, muscle water permeability and function were compared in wild-type and AQP4 knockout mice. Immunofluorescence and freeze-fracture electron microscopy showed AQP4 protein expression in plasmalemma of fast-twitch skeletal muscle fibers of wild-type mice. Osmotic water permeability was measured in microdissected muscle fibers from the extensor digitorum longus (EDL) and fractionated membrane vesicles from EDL homogenates. With the use of spatial filtering microscopy to measure osmotically induced volume changes in EDL fibers, half times (t(1/2)) for osmotic equilibration (7.5-8.5 s) were not affected by AQP4 deletion. Stopped-flow light-scattering measurements of osmotically induced volume changes in plasmalemma vesicles also showed no significant differences in water permeability. Similar water permeability, yet approximately 90% decreased AQP4 protein expression was found in EDL from mdx mice that lack dystrophin. Skeletal muscle function was measured by force generation in isolated EDL, treadmill performance time, and in vivo muscle swelling in response to water intoxication. No differences were found in EDL force generation after electrical stimulation [42 +/- 2 (wild-type) vs. 41 +/- 2 (knockout) g/s], treadmill performance time (22 vs. 26 min; 29 m/min, 13 degrees incline), or muscle swelling (2.8 vs. 2.9% increased water content at 90 min after intraperitoneal water infusion). Together these results provide evidence against a significant role of AQP4 in skeletal muscle physiology in mice. PMID- 10837339 TI - Pyrrolidine dithiocarbamate-induced apoptosis depends on cell type, density, and the presence of Cu(2+) and Zn(2+). AB - Pyrrolidine dithiocarbamate (PDTC) has been found to induce or inhibit apoptosis in different cell types. Here we show that PDTC dose-dependently reduced the viability of rat smooth muscle cells (rSMC), human fibroblasts, and endothelial cells at low but not at high cell density. Endothelial cells were least sensitive, fibroblasts showed a medium sensitivity, and rSMC showed a high sensitivity to PDTC-mediated cell death. An early reduction in the mitochondrial membrane potential indicated a rapid onset of apoptosis in rSMC. Apoptosis was further confirmed by annexin V staining and DNA fragmentation analysis. Gel shift analysis demonstrated increased nuclear factor (NF)-kappaB activity in high density rSMC compared with low-density cells. NF-kappaB has recently been shown to regulate the induction of anti-apoptotic proteins. Although PDTC is widely used as an inhibitor for NF-kappaB and a radical scavenger, our data show that PDTC rather enhanced NF-kappaB activity and, alone or in combination with menadione, induced oxygen radical generation. Notably, PDTC failed to reduce rSMC viability in medium without Cu(2+) or Zn(2+), and addition of Cu(2+) or Zn(2+) resulted in a dose-dependent increase in PDTC-induced cell death. Addition of both Cu(2+) and Zn(2+) showed synergistic effects. Our results indicate that the induction of apoptosis by PDTC requires Cu(2+) and Zn(2+) and is dependent on cell type and density. Such differential effects may have implications for studies of PDTC as an anti-atherosclerotic or immunomodulatory drug. PMID- 10837340 TI - Mechanical strain inhibits expression of osteoclast differentiation factor by murine stromal cells. AB - Normal dynamic loading prevents bone resorption; however, the means whereby biophysical factors reduce osteoclast activity are not understood. We show here that mechanical strain (2% at 10 cycles per minute) applied to murine marrow cultures reduced 1, 25(OH)(2)D(3)-stimulated osteoclast formation by 50%. This was preceded by decreased expression of osteoclast differentiation factor (ODF/TRANCE). RT-PCR for ODF/TRANCE revealed that ODF/TRANCE mRNA in strained cultures was 59 +/- 3% of that seen in control cultures. No significant effects on total cell count, thymidine uptake, or alkaline phosphatase activity were induced by strain. To isolate the cell targeted by strain, primary stromal cells were cultured from marrow. Mechanical strain also reduced mRNA for ODF/TRANCE to 60% that of control in these cells. In contrast, mRNA for membrane-bound macrophage colony-stimulating factor was not significantly affected. Soluble ODF ( approximately 2 ng/ml) was able to reverse the effect of strain, returning osteoclast numbers to control. Because osteoclast formation is dependent upon ODF/TRANCE expression, strain-induced reductions in this factor may contribute to the accompanying reduction in osteoclastogenesis. PMID- 10837341 TI - Expression of smooth muscle myosin light chain 17 and unloaded shortening in single smooth muscle cells. AB - These experiments were performed to test the hypotheses that myosin light chain 17 (MLC(17)) a and b isoform expression varies between individual vascular smooth muscle (SM) cells and that their expression correlates with cell unloaded shortening velocity. Single SM cells isolated from rabbit aorta and carotid arteries were used to measure unloaded shortening velocity and subsequently were analyzed via RT-PCR for MLC(17) a and b mRNA ratio. The MLC(17b/a) mRNA and protein ratios from adjacent tissue sections correlate very well (R(2) = 0.68), allowing use of the mRNA ratio to predict the protein ratio. The rabbit MLC(17) isoform protein sequence was found to be similar to, but unique from, the swine, mouse, and chicken sequences. Isolated single SM cells from the aorta and carotid have resting lengths of 70-280 microm and shorten to 33-88 microm after contraction. Isolated cell maximum unloaded shortening velocity is highly variable (0.5-7.5 microm/s) but becomes more uniform when normalized to initial cell length (0.01-0.05 cell lengths/s). Carotid cells activated in the presence of okadaic acid (1 microm) have mean maximal unloaded shortening velocities not significantly different from carotid cells activated without okadaic acid (0.016 vs. 0.019 cell lengths/s). Resting cell length before activation is significantly correlated with final cell length after unloaded shortening. Neither initial cell length, final cell length, total cell length change, nor maximum unloaded shortening velocity (absolute or normalized) was significantly correlated with single-cell MLC(17b/a) mRNA ratio. These studies were performed in isolated single SM cells where unloaded shortening velocity and MLC(17b/a) mRNA ratios were measured in the same cell. In this preparation, the three-dimensional organization and milieu of the cell is kept intact, but without the intercellular heterogeneity concerns of multicellular preparations. These results suggest the MLC(17b/a) ratio is variable between individual SM cells from the same tissue, but it is not a determinant of unloaded shortening velocity in single SM cells. PMID- 10837342 TI - Limited myogenic response to a single bout of weight-lifting exercise in old rats. AB - The purpose of the present study was to compare the myogenic response of hindlimb muscles in young (14-20 wk of age) and old (>120 wk of age) rats with a single exhaustive bout of heavy resistance weight lifting. [(3)H]thymidine and [(14)C]leucine labeling were monitored for up to 2 wk after the exercise bout to estimate serial changes in mitotic activity and the level of amino acid uptake and myosin synthesis. Histological, histochemical, and immunohistochemical [anti 5-bromo-2'-deoxyuridine and myogenic determination genes (MyoD)] analyses of whole muscles and analysis of muscle-specific gene expression (MyoD) using Western blotting and RT-PCR were performed. Old rats showed significant muscle atrophy and a lower exercise capacity than young rats. Exercise-induced muscle damage, as assessed in histological sections, and increases in serum creatine kinase activity were evident in both young and old exercised groups. Mitotic activity was increased in young, but not old, rats 2 days after exercise. There was a biphasic increase in [(14)C]leucine uptake during the 14 days postexercise (peaks at 1-4 and 10 days) in young rats: only the first peak was observed in old rats. There was a lower uptake of [(14)C]leucine in the myosin fraction and an impaired expression of MyoD at the protein (immunohistochemistry and Western blotting) and mRNA (RT-PCR) levels in old rats throughout the postexercise period. These results demonstrate a reduced reparative capability of muscle in response to a single bout of exercise in old compared with young rats. PMID- 10837343 TI - In vivo regulation of the beta-myosin heavy chain gene in soleus muscle of suspended and weight-bearing rats. AB - In the weight-bearing hindlimb soleus muscle of the rat, approximately 90% of muscle fibers express the beta-myosin heavy chain (beta-MHC) isoform protein. Hindlimb suspension (HS) causes the MHC isoform population to shift from beta toward the fast MHC isoforms. Our aim was to establish a model to test the hypothesis that this shift in expression is transcriptionally regulated through specific cis elements of the beta-MHC promoter. With the use of a direct gene transfer approach, we determined the activity of different length beta-MHC promoter fragments, linked to a firefly luciferase reporter gene, in soleus muscle of control and HS rats. In weight-bearing rats, the relative luciferase activity of the longest beta-promoter fragment (-3500 bp) was threefold higher than the shorter promoter constructs, which suggests that an enhancer sequence is present in the upstream promoter region. After 1 wk of HS, the reporter activities of the -3500-, -914-, and -408-bp promoter constructs were significantly reduced ( approximately 40%), compared with the control muscles. However, using the -215-bp construct, no differences in promoter activity were observed between HS and control muscles, which indicates that the response to HS in the rodent appears to be regulated within the -408 and -215 bp of the promoter. PMID- 10837344 TI - Development and polarization of cationic amino acid transporters and regulators in the human placenta. AB - We have investigated L-arginine transport systems in the human placental syncytiotrophoblast across gestation using purified microvillous (MVM) and basal (BM) plasma membrane vesicles. In MVM from first-trimester and term placentas, L arginine transport was by systems y(+) and y(+)L. In BM (term placentas), however, there was evidence for system y(+)L only. The Michaelis constant of system y(+)L was significantly lower (P < 0.05) in first-trimester compared with term MVM and lower in term MVM compared with BM (P < 0.05). There was no functional evidence for system b(0+) in term MVM or BM. Cationic amino acid transporter (CAT) 1, CAT 4, and 4F2hc were detected using RT-PCR in placentas throughout gestation. rBAT was not detected in term placentas. An approximately 85-kDa and an approximately 135-kDa protein was detected by Western blotting in MVM under reducing and nonreducing conditions, respectively, consistent with the 4F2hc monomer and the 4F2hc-light chain dimer, and their expression was significantly higher (P < 0.05) in term compared with first-trimester MVM. These proteins were not detected in BM despite functional evidence for system y(+)L. These data suggest different roles for 4F2hc in the development and polarization of cationic amino acid transporters in the syncytiotrophoblast. PMID- 10837345 TI - 150-kDa oxygen-regulated protein (ORP150) functions as a novel molecular chaperone in MDCK cells. AB - To assess the participation of the 150-kDa oxygen-regulated protein (ORP150) in protein transport, its function in Madin-Darby canine kidney (MDCK) cells was studied. Exposure of MDCK cells to hypoxia resulted in an increase of ORP150 antigen and increased binding of ORP150 to GP80/clusterin (80-kDa glycoprotein), a natural secretory protein in this cell line. In ORP150 antisense transformant MDCK cells, GP80 was retained within the endoplasmic reticulum after exposure to hypoxia. Metabolic labeling showed the delay of GP80 maturation in antisense transformants in hypoxia, whereas its matured form was detected in wild-type cells, indicating a role of ORP150 in protein transport, especially in hypoxia. The affinity chromatographic analysis of ORP150 suggested its ability to bind to ATP-agarose. Furthermore, the ATP hydrolysis analysis showed that ORP150 can release GP80 at a lower ATP concentration. These data indicate that ORP150 may function as a unique molecular chaperone in renal epithelial cells by facilitating protein transport/maturation in an environment where less ATP is accessible. PMID- 10837346 TI - Separate entry pathways for phosphate and oxalate in rat brain microsomes. AB - ATP-dependent (45)Ca uptake in rat brain microsomes was measured in intracellular like media containing different concentrations of PO(4) and oxalate. In the absence of divalent anions, there was a transient (45)Ca accumulation, lasting only a few minutes. Addition of PO(4) did not change the initial accumulation but added a second stage that increased with PO(4) concentration. Accumulation during the second stage was inhibited by the following anion transport inhibitors: niflumic acid (50 microM), 4,4'-dinitrostilbene-2, 2'-disulfonic acid (DNDS; 250 microM), and DIDS (3-5 microM); accumulation during the initial stage was unaffected. Higher concentrations of DIDS (100 microM), however, inhibited the initial stage as well. Uptake was unaffected by 20 mM Na, an activator, or 1 mM arsenate, an inhibitor of Na-PO(4) cotransport. An oxalate-supported (45)Ca uptake was larger, less sensitive to DIDS, and enhanced by the catalytic subunit of protein kinase A (40 U/ml). Combinations of PO(4) and oxalate had activating and inhibitory effects that could be explained by PO(4) inhibition of an oxalate dependent pathway, but not vice versa. These results support the existence of separate transport pathways for oxalate and PO(4) in brain endoplasmic reticulum. PMID- 10837347 TI - Bioactive products of arginine in sepsis: tissue and plasma composition after LPS and iNOS blockade. AB - Blockade or gene deletion of inducible nitric oxide synthase (iNOS) fails to fully abrogate all the sequelae leading to the high morbidity of septicemia. An increase in substrate uptake may be necessary for the increased production of nitric oxide (NO), but arginine is also a precursor for other bioactive products. Herein, we demonstrate an increase in alternate arginine products via arginine and ornithine decarboxylase in rats given lipopolysaccharide (LPS). The expression of iNOS mRNA in renal tissue was evident 60 but not 30 min post-LPS, yet a rapid decrease in blood pressure was obtained within 30 min that was completely inhibited by selective iNOS blockade. Plasma levels of arginine and ornithine decreased by at least 30% within 60 min of LPS administration, an effect not inhibited by the iNOS blocker L-N(6)(1-iminoethyl)lysine (L-NIL). Significant increases in plasma nitrates and citrulline occurred only 3-4 h post LPS, an effect blocked by L-NIL pretreatment. The intracellular composition of organs harvested 6 h post-LPS reflected tissue-specific profiles of arginine and related metabolites. Tissue arginine concentration, normally an order of magnitude higher than in plasma, did not decrease after LPS. Pretreatment with L NIL had a significant impact on the disposition of tissue arginine that was organ specific. These data demonstrate changes in arginine metabolism before and after de novo iNOS activity. Selective blockade of iNOS did not prevent uptake and can deregulate the production of other bioactive arginine metabolites. PMID- 10837348 TI - An electrogenic Na(+)-HCO(-)(3) cotransporter (NBC) with a novel COOH-terminus, cloned from rat brain. AB - We screened rat brain cDNA libraries and used 5' rapid amplification of cDNA ends to clone two electrogenic Na(+)-HCO(-)(3) cotransporter (NBC) isoforms from rat brain (rb1NBC and rb2NBC). At the amino acid level, one clone (rb1NBC) is 96% identical to human pancreas NBC. The other clone (rb2NBC) is identical to rb1NBC except for 61 unique COOH-terminal amino acids, the result of a 97-bp deletion near the 3' end of the open-reading frame. Using RT-PCR, we confirmed that mRNA from rat brain contains this 97-bp deletion. Furthermore, we generated rabbit polyclonal antibodies that distinguish between the unique COOH-termini of rb1NBC (alpharb1NBC) and rb2NBC (alpharb2NBC). alpharb1NBC labels an approximately 130 kDa protein predominantly from kidney, and alpharb2NBC labels an approximately 130-kDa protein predominantly from brain. alpharb2NBC labels a protein that is more highly expressed in cortical neurons than astrocytes cultured from rat brain; alpharb1NBC exhibits the opposite pattern. In expression studies, applying 1.5% CO(2)/10 mM HCO(-)(3) to Xenopus oocytes injected with rb2NBC cRNA causes 1) pH(i) to recover from the initial CO(2)-induced acidification and 2) the cell to hyperpolarize. Subsequently, removing external Na(+) reverses the pH(i) increase and elicits a rapid depolarization. In the presence of 450 microM DIDS, removing external Na(+) has no effect on pH(i) and elicits a small hyperpolarization. The rate of the pH(i) decrease elicited by removing Na(+) is insensitive to removing external Cl(-). Thus rb2NBC is a DIDS-sensitive, electrogenic NBC that is predominantly expressed in brain of at least rat. PMID- 10837349 TI - The cGMP-dependent protein kinase stimulates the basolateral 18-pS K channel of the rat CCD. AB - We used the patch-clamp technique to study the effect of cGMP on the 18-pS K channel in the basolateral membrane of the rat cortical collecting duct. Addition of 100 microM 8-bromoguanosine 3', 5'-cyclic monophosphate (8-Br-cGMP) increased the activity of the 18-pS K channel, defined by NP(o), by 95%. In contrast, applying 8-bromoadenosine 3',5'-cyclic monophosphate (8-Br-cAMP) has no effect on channel activity. The effect of 8-Br-cGMP was observed only in cell-attached but not in inside-out patches. Application of 1 microM KT-5823, an inhibitor of the cGMP-dependent protein kinase (PKG), not only reduced the channel activity, but also completely abolished the stimulatory effect of 8-Br-cGMP, suggesting that the 18-pS K channel is not a cGMP-gated K channel. Addition of H-89, an agent that also blocks the PKG, mimicked the effect of KT-5823. To examine the possibility that the effect of 8-Br-cGMP is the result of inhibiting cGMP dependent phosphodiesterase (PDE) and, accordingly, increasing cAMP or cGMP levels, we explored the effect on the 18-pS K channel of IBMX, an agent that inhibits the PDE. The addition of 100 microM IBMX had no significant effect on channel activity in cell-attached patches. Moreover, in the presence of IBMX, 8 Br-cGMP increased the channel activity to the same extent as that observed in the absence of IBMX, suggesting that the effect of cGMP is not mediated by inhibiting the cGMP-dependent PDE. That the effect of cGMP is mediated by stimulating PKG was further indicated by experiments in which application of exogenous PKG restored the channel activity when it decreased after the excision of the patches. In contrast, adding exogenous cAMP-dependent protein kinase catalytic subunit failed to reactivate the run-down channels. We conclude that cGMP stimulates the 18-pS channel, and the effect of cGMP is mediated by PKG. PMID- 10837350 TI - Extracellular Cl(-) modulates shrinkage-induced activation of Na(+)/H(+) exchanger in rat mesangial cells. AB - To examine the effect of hyperosmolality on Na(+)/H(+) exchanger (NHE) activity in mesangial cells (MCs), we used a pH-sensitive dye, 2',7'-bis(2-carboxyethyl) 5(6)-carboxyfluorescein-AM, to measure intracellular pH (pH(i)) in a single MC from rat glomeruli. All the experiments were performed in CO(2)/HCO(-)(3)-free HEPES solutions. Exposure of MCs to hyperosmotic HEPES solutions (500 mosmol/kgH(2)O) treated with mannitol caused cell alkalinization. The hyperosmolality-induced cell alkalinization was inhibited by 100 microM ethylisopropylamiloride, a specific NHE inhibitor, and was dependent on extracellular Na(+). The hyperosmolality shifted the Na(+)-dependent acid extrusion rate vs. pH(i) by 0.15-0.3 pH units in the alkaline direction. Removal of extracellular Cl(-) by replacement with gluconate completely abolished the rate of cell alkalinization induced by hyperosmolality and inhibited the Na(+) dependent acid extrusion rate, whereas, under isosmotic conditions, it caused no effect on Na(+)-dependent pH(i) recovery rate or Na(+)-dependent acid extrusion rate. The Cl(-)-dependent cell alkalinization rate under hyperosmotic conditions was partially inhibited by pretreatment with 5-nitro-2-(3 phenylpropylamino)benzoic acid, DIDS, and colchicine. We conclude: 1) in MCs, hyperosmolality activates NHE to cause cell alkalinization, 2) the acid extrusion rate via NHE is greater under hyperosmotic conditions than under isosmotic conditions at a wide range of pH(i), 3) the NHE activation under hyperosmotic conditions, but not under isosmotic conditions, requires extracellular Cl(-), and 4) the Cl(-)-dependent NHE activation under hyperosmotic conditions partly occurs via Cl(-) channel and microtubule-dependent processes. PMID- 10837351 TI - Polarity of A2b adenosine receptor expression determines characteristics of receptor desensitization. AB - It is not known if, in polarized cells, desensitization events can be influenced by the domain on which the receptor resides. Desensitization was induced by 5'-(N ethylcarboxamido)adenosine (NECA) and was quantitated by measurement of short circuit current (I(sc)) in response to adenosine. NECA added to either the apical or basolateral compartments rapidly desensitized receptors on these respective domains. Although apical NECA had no effect on the basolateral receptor stimulation, basolateral NECA induced a complete desensitization of the apical receptor. We hypothesized that desensitization of apical receptor by basolateral desensitization could relate to a trafficking step in which A2b receptor is first targeted basolaterally upon synthesis and transported to the apical surface via vesicular transport/microtubules. Because desensitization is associated with downregulation of receptors, apical adenosine receptor can thus be affected by basolateral desensitization. Both low temperature and nocodazole inhibited I(sc) induced by apical and not basolateral adenosine. IN CONCLUSION: 1) a single receptor subtype, here modeled by the A2b receptor, differentially desensitizes based on the membrane domain on which it is expressed, 2) agonist exposure on one domain can result in desensitization of receptors on the opposite domain, 3) cross-domain desensitization can display strict polarity, and 4) receptor trafficking may play a role in the cross-desensitization process. PMID- 10837352 TI - Effects of altered tonicity by sodium chloride on L-tryptophan binding to hepatic nuclei. AB - This study was concerned with the effects of NaCl administered in vivo or added in vitro to isolated nuclei on [(3)H]tryptophan binding to rat hepatic nuclei assayed in vitro. Hypertonic (10.7%) NaCl administered in vivo to rats caused at 10 min a marked decrease in in vitro binding (total and specific) of [(3)H]tryptophan to hepatic nuclei. In vitro incubation of isolated hepatic nuclei, but not of isolated nuclear envelopes, with added NaCl (particularly at 0.125 x 10(-4) M and 0.25 x 10(-4) M) revealed significant inhibition of [(3)H]tryptophan binding. However, isolated hepatic nuclear envelopes prepared after in vitro incubation of isolated nuclei with added NaCl did show inhibition of [(3)H]tryptophan binding (total and specific) compared with controls. Other salts (KCl, MgCl(2), NaHCO(3), NaC(2)H(3)O(2), NaF, or Na(2)SO(4)), at similar concentrations to that of NaCl except for MgCl(2), when added to isolated nuclei did not appreciably inhibit nuclear tryptophan binding. Kinetic studies of in vitro nuclear [(3)H]tryptophan binding in the presence of 0.125 x 10(-4) M NaCl revealed that binding decreased at 0.5 h and continued to 2 h compared with nuclear [(3)H]tryptophan binding with controls (without NaCl addition). The results obtained in vivo in rats and those obtained in vitro with isolated hepatic nuclei revealed NaCl-induced inhibitory effects on [(3)H]tryptophan binding to hepatic nuclei. Although the inhibitory effects were similar under the two different experimental conditions, the mechanism for each may be different in that the NaCl concentration in hepatic cells after administration of NaCl in vivo was appreciably higher than the low levels added in vitro to the isolated hepatic nuclei. PMID- 10837353 TI - The human histamine H(2) receptor regulates c-jun and c-fos in a differential manner. AB - Previously, we demonstrated that activation of the human H(2) receptor (hH(2)R) leads to an increase in c-fos transcription and cell proliferation. The purpose of these studies was to examine whether hH(2)R regulates c-jun expression and, if so, explore the mechanisms by which it does so. Histamine induced an increase in c-jun mRNA in human embryonic kidney cells stably transfected with the hH(2)R (maximal effect: 554.6 +/- 86.8% of control). The protein kinase C (PKC) inhibitors staurosporine (10(-6) M) and GF-109203X (10(-6) M) significantly inhibited histamine-stimulated c-fos mRNA while not altering c-jun expression. The protein kinase A (PKA) pathway inhibitors Rp-cAMP and protein kinase inhibitor did not affect the action of histamine on c-jun or c-fos mRNA. Histamine (10(-4) M) stimulated extracellularly regulated kinase 2 tyrosine phosphorylation. The specific inhibitor of the mitogen-activated protein (MAP) kinase pathway, PD-98059 (5 x 10(-5) M), significantly inhibited histamine induced c-fos and c-jun mRNA. Of interest, the p70 S6 kinase inhibitor rapamycin (10(-6) M) but not wortmannin decreased histamine-stimulated c-jun mRNA by 58.5 +/- 12% (mean +/- SE, n = 4) while not significantly altering c-fos message. Histamine (10(-4) M) also led to an approximately 4.5-fold increase in Jun NH(2) terminal kinase activity in a PKC-, PKA-, and MAP kinase-independent but rapamycin-sensitive manner. Our findings suggest that histamine stimulates both c fos and c-jun mRNA in a differential manner. PKC is involved in histamine mediated c-fos activation, whereas p70 S6 kinase is important for linkage of this receptor to c-jun. PMID- 10837354 TI - Role of multidrug resistance P-glycoprotein in the secretion of aldosterone by human adrenal NCI-H295 cells. AB - We determined the role of the multidrug resistance (MDR1) gene product, P glycoprotein (PGP), in the secretion of aldosterone by the adrenal cell line NCI H295. Aldosterone secretion is significantly decreased by the PGP inhibitors verapamil, cyclosporin A (CSA), PSC-833, and vinblastine. Aldosterone inhibits the efflux of the PGP substrate rhodamine 123 from NCI-H295 cells and from human mesangial cells (expressing PGP). CSA, verapamil, and the monoclonal antibody UIC2 significantly decreased the efflux of fluorescein-labeled (FL)-aldosterone microinjected into NCI-H295 cells. In MCF-7/VP cells, expressing multidrug resistance-associated protein (MRP) but not PGP, and in the parental cell line MCF7 (expressing no MRP and no PGP), the efflux of microinjected FL-aldosterone was slow. In BC19/3 cells (MCF7 cells transfected with MDR1), the efflux of FL aldosterone was rapid and it was inhibited by verapamil, indicating that transfection with MDR1 cDNA confers the ability to transport FL-aldosterone. These results strongly indicate that PGP plays a role in the secretion of aldosterone by NCI-H295 cells and in other cells expressing MDR1, including normal adrenal cells. PMID- 10837355 TI - Skeletal muscle gene transfer: regeneration-associated deregulation of fast troponin I fiber type specificity. AB - Direct gene transfer into skeletal muscle in vivo presents a convenient experimental approach for studies of adult muscle gene regulatory mechanisms, including fast vs. slow fiber type specificity. Previous studies have reported preferential expression of fast myosin heavy chain and slow myosin light chain and troponin I (TnIslow) gene constructs in muscles enriched in the appropriate fiber type. We now report a troponin I fast (TnIfast) direct gene transfer study. We injected into the mouse soleus muscle plasmid DNA or recombinant adenovirus carrying a TnIfast/ beta-galactosidase (beta-gal) reporter construct that had previously been shown to be expressed specifically in fast fibers in transgenic mice. Surprisingly, microscopic histochemical analysis 1 and 4 wk postinjection showed similar TnIfast/beta-gal expression in fast and slow fibers. A low but significant level of muscle fiber segmental regeneration was evident in muscles 1 wk postinjection, and TnIfast/beta-gal expression was preferentially targeted to regenerating fiber segments. This finding can explain why TnIfast constructs are deregulated with regard to fiber type specificity, whereas the myosin constructs previously studied are not. The involvement of regenerating fiber segments in transduction by plasmid DNA and recombinant adenoviruses injected into intact normal adult muscle is an unanticipated factor that should be taken into account in the planning and interpretation of direct gene transfer experiments. PMID- 10837356 TI - Trophic slime, allergic slime. PMID- 10837357 TI - Gene therapy for pulmonary edema. PMID- 10837358 TI - Lactoperoxidase. New recognition of an "old" enzyme in airway defenses. PMID- 10837359 TI - Case-control association studies for the genetics of complex respiratory diseases. PMID- 10837360 TI - Interleukin-9 upregulates mucus expression in the airways. AB - Interleukin (IL)-9 has recently been shown to play an important role in allergic disease because its expression is strongly associated with the degree of airway responsiveness and the asthmatic-like phenotype. IL-9 is a pleiotropic cytokine that is active on many cell types involved in the allergic immune response. Mucus hypersecretion is a clinical feature of chronic airway diseases; however, the mechanisms underlying the induction of mucin are poorly understood. In this report, we show that IL-9 regulates the expression of a subset of mucin genes in lung cells both in vivo and in vitro. In vivo, the constitutive expression of IL 9 in transgenic mice results in elevated MUC2 and MUC5AC gene expression in airway epithelial cells and periodic acid-Schiff-positive staining (reflecting mucous glycogenates). Similar results were observed in C57BL/6J mice after IL-9 intratracheal instillation. In contrast, instillation of the T helper 1 associated cytokine interferon gamma failed to induce mucin production. In vitro, our studies showed that IL-9 also induces expression of MUC2 and MUC5AC in human primary lung cultures and in the human muccoepidermoid NCI-H292 cell line, indicating a direct effect of IL-9 on inducing mucin expression in these cells. Altogether, these results suggest that upregulation of mucin by IL-9 might contribute to the pathogenesis of human inflammatory airway disorders, such as asthma. These data extend the role of the biologic processes that IL-9 has on regulating the many clinical features of asthma and further supports the IL-9 pathway as a key mediator of the asthmatic response. PMID- 10837361 TI - Lung overexpression of the vascular endothelial growth factor gene induces pulmonary edema. AB - We hypothesized that the angiogenic mediator, vascular endothelial growth factor (VEGF), known to be expressed in the lung and to be capable of inducing local edema in skin, might evoke the development of lung edema if expressed in excess amounts. To test this hypothesis, we developed an in vivo model of VEGF overexpression in the lung on the basis of delivery to the respiratory epithelium of the VEGF165 complementary DNA by an E1(-) adenovirus vector (AdVEGF165). Administration of AdVEGF165 by the intratracheal route (10(9) plaque-forming units [pfu]) to C57Bl/6 mice showed increased expression of VEGF messenger RNA in lung tissue by Northern analysis. Overexpression of VEGF protein in the lung at Days 1 to 10 was confirmed by enzyme-linked immunosorbent assay. Intratracheal administration of AdVEGF165 resulted in a dose-dependent increase in lung wet/dry weight ratios over time, lung histology showed widespread intra- alveolar edema, and pulmonary capillary permeability was significantly increased as quantified by the Evans blue dye assay and [(131)I]albumin permeability. To confirm the specificity of these observations, mice were pretreated with intranasal administration of an adenovirus vector expressing a truncated soluble form of the VEGF receptor flt-1 (Adsflt). Adsflt (10(9) pfu) pretreatment completely abrogated the increased lung wet/dry weight ratio caused by AdVEGF165 administration, whereas an identical adenovirus vector with an irrelevant transgene had no effect upon subsequent AdVEGF165-induced pulmonary edema. Together, these data suggest that overexpression of VEGF in the lung may be one mechanism of increased pulmonary vascular permeability in the early stages of acute lung injury. PMID- 10837362 TI - The lactoperoxidase system functions in bacterial clearance of airways. AB - Airway mucus is a complex mixture of secretory products that provides a multifaceted defense against pulmonary infection. Mucus contains antimicrobial peptides (e.g., defensins) and enzymes (e.g., lysozyme) although the contribution of these to airway sterility has not been tested in vivo. We have previously shown that an enzymatically active, heme-containing peroxidase comprises 1% of the soluble protein in sheep airway secretions, and it has been hypothesized that this airway peroxidase may function as a biocidal system. In this study, we show that sheep airway peroxidase is identical to milk lactoperoxidase (LPO) and that sheep airway secretions contain thiocyanate (SCN(-)) at concentrations necessary and sufficient for a functional peroxidase system that can protect against infection. We also show that airway LPO, like milk LPO, produces the biocidal compound hypothiocyanite (OSCN(-)) in vitro. Finally, we show that in vivo inhibition of airway LPO in sheep leads to a significant decrease in bacterial clearance from the airways. The data suggest that the LPO system is a major contributor to airway defenses. This discovery may have significant implications for chronic airway colonization seen in respiratory diseases such as cystic fibrosis. PMID- 10837363 TI - The natural resistance-associated macrophage protein gene in African Americans with sarcoidosis. AB - The histologic and clinical similarities between tuberculosis and sarcoidosis suggest a shared underlying pathophysiology. Human natural resistance-associated macrophage protein (NRAMP1), which is closely related to the mouse gene, has been associated with susceptibility to tuberculosis in some human populations. Given the importance of the Nramp1 gene in animal models of granulomatous disorders, the association with human tuberculosis, and the possible role of NRAMP1 in macrophage activation and function, we hypothesized that human NRAMP1 plays a role in susceptibility to sarcoidosis. We analyzed several NRAMP1 gene polymorphisms in a case-control study of 157 African American patients with sarcoidosis and 111 African American control subjects. Our results, in contrast to those in tuberculosis patients, showed that the less common genotypes were found more often in control subjects than in case patients (odds ratio, 0.48; 95% confidence interval, 0.28-0.81). In particular, one polymorphism, a (CA)(n) repeat in the immediate 5' region of the gene, was found to have a protective effect (P = 0. 014). Whereas NRAMP1 polymorphisms have been associated with increased susceptibility to tuberculosis, our results suggest that at least one NRAMP1 polymorphism may decrease susceptibility in sarcoidosis. PMID- 10837364 TI - Antisense inhibition of epidermal growth factor receptor decreases expression of human surfactant protein A. AB - Epidermal growth factor (EGF) stimulates surfactant protein A (SP-A) synthesis in fetal lung tissue through ligand binding to the EGF receptor. We hypothesized that inhibition of EGF receptor messenger RNA (mRNA) would block SP-A expression in human fetal lung tissue during alveolar type II cell differentiation in vitro. Midtrimester human fetal lung explants were maintained in serum-free Waymouth's medium for 3 to 5 d in the presence or absence of an antisense 18-mer phosphorothioate oligonucleotide (ON) complementary to the initiation codon region of EGF receptor mRNA. Sense and scrambled ONs similarly modified were used as additional controls. The concentration of EGF receptor mRNA was semiquantitatively determined by reverse transcriptase/polymerase chain reaction (RT-PCR). We found a significant 3-fold decrease in EGF receptor mRNA levels in the antisense-treated groups compared with the control group with no effect in the sense condition. Immunohistochemical staining revealed a decrease in the amount of staining for EGF receptor protein in distal pulmonary epithelial cells in the antisense-treated groups compared with either control or sense conditions. Treatment with antisense EGF receptor ON decreased both SP-A mRNA and protein compared with controls with no effect in the sense condition. The ONs did not affect tissue viability as measured by the release of lactate dehydrogenase. We conclude that selective degradation of EGF receptor mRNA with antisense ON treatment results in a decrease in SP-A expression in human fetal lung. These findings support the critical importance of the EGF receptor for the regulation of SP-A gene expression during human alveolar type II cell differentiation. PMID- 10837365 TI - Effects of TNF-alpha on expression of ICAM-1 in human airway epithelial cells in vitro. Signaling pathways controlling surface and gene expression. AB - Signaling pathways associated with tumor necrosis factor (TNF)-alpha-induced intercellular adhesion molecule 1 (ICAM-1) surface and gene expression were investigated in well differentiated normal human bronchial epithelial (NHBE) cells in air-liquid interface primary culture. Cells were exposed to human recombinant TNF-alpha (hrTNF-alpha; 0.015 to 150 ng/ml [specific activity, 2.86 x 10(7) U/mg]). TNF-alpha enhanced ICAM-1 surface expression (measured by flow cytometry) and steady-state messenger RNA (mRNA) levels (assessed by Northern hybridization) in concentration- and time-dependent manners. TNF-alpha-induced ICAM-1 surface and gene expression were both blocked by the RNA polymerase II inhibitor actinomycin D (0.1 microg/ml), and surface expression was attenuated by a neutralizing monoclonal antibody directed against the TNF-alpha receptor p55 (TNF-RI). The intracellular signaling pathway leading to enhanced expression appeared to involve activation of a phospholipase C that hydrolyzes phosphatidylcholine (PC-PLC) because D609, a specific PC-PLC inhibitor, attenuated TNF-alpha-induced increases in production of diacyl-glycerol (DAG), a hydrolysis product of PC-PLC, and also attenuated TNF-alpha enhancement of ICAM-1 surface and gene expression. Because DAG formed by action of PC-PLC can activate protein kinase C (PKC), involvement of PKC was investigated. The specific PKC inhibitor calphostin C blocked both surface and gene expression of ICAM-1 in response to TNF-alpha in a concentration-dependent manner. Finally, TNF-alpha stimulated binding of p65 and/or c-rel complexes to the nuclear factor (NF) kappaB consensus binding site found on the ICAM-1 promoter, and binding of these complexes was inhibited by D609. The results support the following pathway, whereby TNF-alpha enhances expression of ICAM-1 in NHBE cells: TNF-alpha --> TNF RI --> PC-PLC --> DAG --> PKC --> (NF-kappaB?) --> ICAM-1 mRNA --> ICAM-1 surface expression. PMID- 10837366 TI - Glucocorticoid receptor activation reduces CD11b and CD49d levels on murine eosinophils: characterization and functional relevance. AB - In vitro incubation of mouse blood eosinophils with dexamethasone (DEX) resulted in concentration- and time-dependent reduction in CD11b and CD49d cell-surface expression as detected by flow cytometry. This inhibitory effect ranged between 20 and 40% for both integrins, and it was not related to alteration of cell survival. DEX was maximally effective at 1 microM, and it was prevented by coaddition of the glucocorticoid receptor antagonist RU486 (mifepristone; 10 microM). Budesonide, hydrocortisone, and prednisolone, but not the sex steroids testosterone and progesterone, reduced CD11b and CD49d cell-surface expression to a similar extent. Subchronic treatment of mice with 1 mg/kg DEX again reduced both CD11b and CD49d expression on circulating eosinophils, without alterations in CD11b messenger RNA expression as assessed by polymerase chain reaction analysis. In contrast, membrane but not intracellular protein expression of either CD11b or CD49d was inhibited by eosinophil incubation with DEX in vitro; thus, an interference with exportation of these adhesion molecules to the cell surface is proposed as the mechanism of action of the glucocorticoid. Finally, steroid effects on integrin expression were linked to a reduced eosinophil function as indicated by a lower degree of cell chemotaxis after incubation with DEX, an effect which was again prevented by 10 microM RU486. These observations may explain part of the therapeutic efficacy displayed by glucocorticoid hormones in the clinical control of tissue eosinophilia in allergic disease conditions. PMID- 10837367 TI - Modification of type I collagenous gels by alveolar epithelial cells. AB - Contraction of type I collagen gels is an in vitro model of tissue remodeling. In addition to fibroblasts, some epithelial cells can mediate this process. We therefore hypothesized that alveolar epithelial cells might contract extracellular matrices and have the potential to directly participate in the remodeling of the lung after alveolar injury. A549 cells were plated on top of collagen gels, and the gels were floated in culture medium. A549 cells contracted the gels in a time- and cell density-dependent manner. A549 cells, as well as human bronchial epithelial cells (HBEC) and rat alveolar epithelial cells (RalvEC) contracted collagen gels more when they were plated on top of the gel than when they were embedded inside, in contrast to human fetal lung fibroblast (HFL1), which contracted more when cast inside. The amount of hydroxyproline in the collagen gels remained unchanged throughout the contraction. Anti-beta(1) integrin antibody inhibited A549 cell-mediated contraction. Transforming growth factor beta augmented the contraction by A549 cells as well as that by HBEC and HFL1. Prostaglandin E(2) inhibited the contraction by HFL1 but did not affect the contraction by A549 cells, HBEC, or RalvEC. Cytomix (a mixture of tumor necrosis factor-alpha, interleukin-1beta, and interferon-gamma) inhibited the contraction by HFL1 but strongly enhanced the contraction by A549 cells. Cytomix also caused a morphologic change of A549 cells from a polygonal to a spindle shape. Immunocytochemistry showed that cytomix induced alpha-tubulin expression in A549 cells, whereas cytokeratin, vimentin, smooth muscle actin, beta(1) integrin, and paxillin expressions were not changed. This study thus demonstrates that alveolar epithelial cells can cause contraction of extracellular matrices and that this process is modulated by exogenous mediators, which also modify the microtubular system. Such an activity might contribute to alveolar remodeling after injury. PMID- 10837368 TI - Expression of interleukin-18 in the lung after endotoxemia or hemorrhage-induced acute lung injury. AB - Hemorrhage and endotoxemia are important risk factors for the development of acute lung injury. Interleukin (IL)-18 is a recently described cytokine released in its mature, active form after pro-IL-18 is cleaved by the IL-1 converting enzyme (ICE). IL-18 has multiple immunomodulating properties, including induction of interferon-gamma (IFN-gamma), IL-1beta, tumor necrosis factor-alpha, and intercellular adhesion molecule-1. To examine the possible involvement of IL-18 in acute lung injury, we examined its expression, as well as that of IFN-gamma, IL-12, and ICE, using murine hemorrhage or endotoxemia models. The amounts of IL 18 messenger RNA (mRNA) increased in the lung after hemorrhage or endotoxemia. However, only endotoxemia was associated with elevations in lung and plasma concentrations of IL-18 protein. ICE expression was increased in the lungs after endotoxemia but not after hemorrhage. Although IFN-gamma expression increased in the lungs after hemorrhage or endotoxemia, elevations in lung IL-12 mRNA levels were found only after endotoxemia. These results indicate that hemorrhage and endotoxemia induce different patterns of immunomodulatory cytokine expression in the lungs. In particular, differences in the expression of ICE after hemorrhage or endotoxemia may affect generation of the active forms of downstream cytokines, including IL-18. IFN-gamma expression in the lungs after hemorrhage appears to occur through a pathway independent of IL-12 and IL-18. IL-18 may play a role in modulating the development of acute lung injury after endotoxemia but not after hemorrhage. PMID- 10837369 TI - Mucoid Pseudomonas aeruginosa, TNF-alpha, and IL-1beta, but not IL-6, induce human beta-defensin-2 in respiratory epithelia. AB - Cultured lung epithelial cells release antibacterial activity upon contact with Pseudomonas aeruginosa (PA), which is impaired in cystic fibrosis (CF). In order to identify the factors responsible for killing PA by a biochemical approach, we purified antimicrobial activity from supernatants of the A549 lung epithelial cell line, previously stimulated with PA bacteria, by subsequent high performance liquid chromatography. NH(2)-terminal sequencing of a major bactericidal compound revealed it to be identical with human beta-defensin-2 (hBD-2). A mucoid phenotype of PA, but not two nonmucoid PA strains, high concentrations (> 10 microg/ml) of PA lipopolysaccharide, tumor necrosis factor alpha, and interleukin (IL)-1beta, but not IL-6, dose-dependently induced hBD-2 messenger RNA in cultured normal bronchial, tracheal, as well as normal and CF-derived nasal epithelial cells. Genomic analysis of hBD-2 revealed a promoter region containing several putative transcription factor binding sites, including nuclear factor (NF) kappaB, activator protein (AP)-1, AP-2, and NF-IL-6, known to be involved in the regulation of inflammatory responses. Thus, hBD-2 represents a major inducible antimicrobial factor released by airway epithelial cells either on contact with mucoid PA or by endogenously produced primary cytokines. Therefore, it might be important in lung infections caused by mucoid PA, including those seen in patients with CF. PMID- 10837370 TI - Pneumocystis carinii uses a functional cdc13 B-type cyclin complex during its life cycle. AB - Pneumocystis carinii causes severe pneumonia in immunocompromised patients. Recent studies indicate that P. carinii uses a Cdc2 cyclin-dependent kinase to control its proliferation. To further study the regulation of the life cycle of P. carinii, we characterized the P. carinii B-type cyclin termed Cdc13, whose binding to Cdc2 is necessary for kinase activity. Antibodies to B-type cyclins (Cdc13) specifically immunoprecipitated Cdc2/ Cdc13 complexes with associated kinase activity from P. carinii extracts. To clone P. carinii cdc13, degenerate polymerase chain reaction was undertaken using primers generated from amino-acid motifs conserved in fungal Cdc13 proteins. This amplicon was used to obtain full length genomic and complementary DNA (cDNA) clones. A specific synthetic peptide antibody generated to P. carinii Cdc13 further demonstrated differential Cdc2/Cdc13 activity over the life cycle of P. carinii, with greater activity in cysts compared with trophic forms of the organism. Finally, P. carinii cdc13 cDNA was used to rescue mutant Schizosaccharomyces pombe strains containing temperature-sensitive deficiencies of endogenous Cdc13 activity, thus verifying function of the P. carinii Cdc13 protein. Therefore, P. carinii contains a Cdc13 cyclin, which is variably active over its life cycle and which promotes fungal proliferation. PMID- 10837371 TI - Attenuation of hyperoxia-induced growth inhibition in H441 cells by gene transfer of mitochondrially targeted glutathione reductase. AB - Reactive oxygen species (ROS) are implicated as agents of cellular damage in pulmonary oxygen toxicity. Glutathione (GSH) and GSH-dependent antioxidant enzymes protect against damage by ROS, and recycling of glutathione disulfide (GSSG) to GSH by glutathione reductase (GR) is essential for the optimum functioning of this system. Exposure to hyperoxia inhibits lung development in newborn animals and humans, and attenuates cell growth in proliferating cell cultures. Considerable evidence supports a role for ROS as growth-altering molecules. Previously, we have observed that gene transfer of GR to mitochondria in H441 cells, using a vector containing a mitochondrial leader sequence (LGR), protected these cells against t-BuOOH-induced cytotoxicity. The present studies tested the hypothesis that gene transfer of LGR would attenuate the cytostatic effects of hyperoxia exposure in H441 cells. H441 cells (0.9 x 10(6) cells/plate) transfected with adenovirus containing LGR or the complementary DNA (cDNA) for manganese superoxide dismutase in reverse orientation (DOS) as a control construct, and untransfected cells (CON) were maintained in 21% oxygen (normoxia) or 95% oxygen (hyperoxia) for 48 h, and cell growth was assessed by cell counts and by reduction of the tetrazolium dye 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) to formazan. Cells maintained in normoxia achieved normal growth (CON, 1.98; DOS, 1.91; LGR, 2.0 x 10(6) cells/plate). Hyperoxia inhibited cell growth and the reduction of MTT; however, cells transfected with LGR had greater mitochondrial GR activities (CON, 16+/-2; DOS, 19+/-3; LGR, 322+/ 18 mU/mg of protein), sustained more normal growth patterns (CON, 1.25+/-0.12; DOS, 1.24 +/-0.21, LGR, 1.8+/-0.25 x 10(6) cells/plate), and had less inhibition of MTT reduction (CON, 29; DOS, 27; LGR, 16% inhibition, P<0.01) after exposure to hyperoxia for 48 h than was observed in cells transfected with DOS or in control cells not infected with virus. In addition, resistant cells had higher mitochondrial GSH levels and maintained mitochondrial GSH/GSSG ratios in hyperoxia, suggesting that maintaining mitochondrial GSH homeostasis determined critical aspects of cell division in these studies. The mechanisms for sustaining cell growth during hyperoxia in H441 cells with enhanced mitochondrial GR activities are unknown, but similar effects in infants exposed to supplemental oxygen could be highly beneficial. PMID- 10837372 TI - Identification of high density lipoprotein-binding proteins, including a glycosyl phosphatidylinositol-anchored membrane dipeptidase, in rat lung and type II pneumocytes. AB - Numerous communications have indicated that specific binding proteins for high density lipoprotein (HDL) exist in addition to the well characterized candidate HDL receptor SR-BI, but structural information was presented only in a few cases, and most of the work was aimed at the liver and steroidogenic glands. In this study, we purified two HDL-binding proteins by standard procedures from rat lung tissue. One of these membrane glycoproteins was identified by N-terminal sequencing and with specific antibodies as HB2, a previously described HDL binding protein, whereas the other one was identified as a glycosyl phosphatidylinositol-anchored membrane dipeptidase (MDP). The apparent dissociation constant of the HDL binding was determined by solid phase assay to be 2.1 microg/ml (HB2) and 25 microg/ml (MDP). MDP also exerts affinity to low density lipoprotein (LDL) on ligand blots, and competition between HDL and LDL was observed, but analysis by solid phase assay showed that very high concentrations of LDL are required. The physiologic relevance of this effect is therefore questionable. The level in type II pneumocyte membranes of both binding proteins, MDP and HB2, increased when the plasma lipoprotein concentration was reduced by treatment of rats with 4-aminopyrazolo[3,4-d]-pyrimidine, consistent with a function to facilitate lipid uptake in vivo. The binding proteins were also dramatically upregulated by feeding rats a vitamin E-depleted diet. Vitamin E uptake requires interaction between HDL and type II cells, suggesting a role of HB2 and MDP also in this process. PMID- 10837374 TI - Change at the BJO PMID- 10837373 TI - Polycyclic aromatic hydrocarbon carcinogens increase ubiquitination of p21 protein after the stabilization of p53 and the expression of p21. AB - Polycyclic aromatic hydrocarbon carcinogens (PAHs) and their metabolites have been found to result in a rapid accumulation of p53 gene product in human and mouse cells. However, the induced p53 protein was reported to be transcriptionally inactive. In the present study, the induction of p53 target gene expression after the treatment with either benzo(a)pyrene (B[a]P) or 1 nitropyrene (1-NP) was investigated. A marked induction of messenger RNA (mRNA) expressions of Mdm2, Bax, and p21 was detected in wild-type p53-expressing cells after the treatment with either B[a]P or 1-NP, whereas no significant change in mRNA expression of these genes was observed in p53-negative and mutant cells. 1 NP activated the p21 promoter in a p53-dependent manner. Binding activity of p53 to a p53 consensus sequence increased after the treatment in wild-type p53 expressing cells. Nevertheless, the induced mRNA levels of the p21 did not result in a proportional p21 protein increase, indicating the possibility of post transcriptional regulation of the protein. With the addition of MG-132, a proteasome inhibitor, to B[a]P or 1-NP treatments, both p21 and p53 protein levels were increased; however, the increase in p21 protein levels was significantly larger than the increase in p53 protein levels. PAHs treatment increased the level of ubiquitinated p21. These results suggest that the p21 product is degraded by the ubiquitin-proteasome system. We conclude that PAHs induced p53 protein is transcriptionally active. PMID- 10837375 TI - Herpes simplex virus in the human cornea. PMID- 10837376 TI - The efficacy of occlusion for strabismic amblyopia. Can an optimal duration be identified? PMID- 10837377 TI - Human herpesviruses in the cornea. AB - AIMS: To determine the sensitivity and specificity of culture, immunohistochemistry (IHC), the polymerase chain reaction (PCR), and in situ hybridisation (ISH) for detecting herpes simplex virus (HSV-1) in the cornea of patients undergoing penetrating keratoplasty. To compare the incidence of HSV-1 in the cornea with that of varicella zoster virus (VZV), cytomegalovirus (CMV), and Epstein-Barr virus (EBV). METHODS: The corneas of 110 patients, 52 with a documented history of herpes keratitis (HSK) and 58 with non-herpetic corneal disease, were investigated using IHC, PCR, ISH, and culture. RESULTS: HSV-1 DNA and antigen were detected in 82% and 74% respectively, of corneas of patients with HSK and in 22% and 15% of corneas of patients with no history of HSK. The sensitivity of PCR and IHC was 82% and 74% with a specificity of 78% and 85%, respectively. HSV-1 DNA and antigen were found more frequently and in increased amounts in corneas of patients with a short interval between their last attack of HSK and surgery. There was a good correlation between PCR and IHC in 71%. HSV-1 was isolated by culture in 2%. Latency associated transcripts were not detected using ISH. Evidence of VZV DNA or antigen was found significantly more frequently in the corneas of patients with a history of HSK (p<0.001). No evidence of EBV or CMV was found in any cornea. CONCLUSIONS: PCR and IHC are both sensitive for the detection of HSV-1 in the cornea. A combination of PCR and IHC increases the specificity for the diagnosis of HSK to 97%. HSV-1 appears to be slowly removed from the cornea. VZV and HSV-1 may co-infect the cornea. PMID- 10837378 TI - Efficacy of occlusion for strabismic amblyopia: can an optimal duration be identified? AB - BACKGROUND/AIMS: The study of occlusion efficacy in amblyopia has been hampered by the use of non-logMAR acuity tests and a failure to assess threshold acuity for both eyes. These issues are addressed in the current study which compares the effect of spectacles alone and spectacles in combination with occlusion, with the use of the logMAR crowded test. METHODS: Changes in uniocular and interocular acuity differences were compared for two age matched groups of previously untreated children with strabismic amblyopia: one compliant with spectacles only (n = 17, mean 6.2 (SD 2.5) years) and the other with spectacles and occlusion (n = 69, mean 5.1 (1) years) over a 1 year period. Changes in logMAR acuity were also analysed for a larger occluded group (n = 119) in response to successive 200 hour blocks of occlusion up to > or =1000 hours, in an attempt to isolate an optimal occlusion regime. RESULTS: Visual acuity improved for more of the amblyopic eyes of the occluded (74%) than the spectacles only group (59%), and only one child from the latter group deteriorated. Mean visual acuity improved for both eyes of both treatment groups, but the change was significantly larger for the strabismic eyes of the occluded group overall and within the first 6 month period (p <0. 05). Occlusion was only effective for the first 400 hours worn. Subsequent visual improvement was bilateral and symmetrical. CONCLUSION: Occlusion is more effective in the treatment of strabismic amblyopia than spectacles alone, and the effect is optimal within the first 6 months of wear. In terms of occlusion duration, maximal improvement occurs in response to 400 hours of occlusion wear or less, and to full time occlusion. Visual maturation continues, but is retarded for amblyopic eyes. PMID- 10837379 TI - A comparison of latanoprost and dorzolamide in patients with glaucoma and ocular hypertension: a 3 month, randomised study. Ireland Latanoprost Study Group. AB - AIMS: To compare the effects on intraocular pressure (IOP) and side effects of monotherapy with either latanoprost or dorzolamide in patients with glaucoma or ocular hypertension. METHODS: 224 patients with open angle glaucoma or ocular hypertension were recruited to a 3 month open labelled study. Previous glaucoma medications were washed out and the patients were randomised to receive either latanoprost 0.005% once daily or dorzolamide 2% three times daily. RESULTS: Of 224 patients 213 were included in the analysis of efficacy. After 3 months, latanoprost reduced mean baseline diurnal IOP from 27.2 (SD 3.0) mm Hg by 8.5 (3.3) mm Hg. The corresponding figures for dorzolamide were 27.2 (3.4) and 5.6 (2.6) mm Hg. The difference of 2.9 mm Hg (95% CI: 2.3-3.6) was highly significant (p<0.001, ANCOVA). Latanoprost reduced IOP at peak by 8.6 mm Hg (32%) compared with 6.2 mm Hg (23%) for dorzolamide, and the difference of 2.4 mm Hg was significant (p<0.001, ANCOVA). The corresponding figures at trough were 8.1 mm Hg (31%) for latanoprost and 4.7 mm Hg (17%) for dorzolamide, a significant difference of 3.4 mm Hg (p<0.001, ANCOVA). Both drugs were well tolerated systemically and locally. CONCLUSION: Latanoprost was superior to dorzolamide in reducing the IOP, judged both from the effect on IOP at peak and trough and by the effect on diurnal IOP. PMID- 10837380 TI - Corneal amyloidosis caused by Leu518Pro mutation of betaig-h3 gene. AB - AIM: To report a Japanese family diagnosed clinically as having lattice corneal dystrophy type I (LCDI) in which a Leu518Pro mutation in the betaig-h3 gene and not the R124C mutation reported previously was found. METHODS: Molecular genetic analysis was performed on DNA extracted from peripheral leucocytes from four members (three affected and one unaffected) of a family. Exon 4 of the betaig-h3 gene was amplified by PCR and directly sequenced. Histopathological study was performed on the corneal tissue from the proband obtained during deep lamellar keratoplasty. RESULTS: All the affected members were clinically diagnosed as having LCDI, and the pedigree indicated an autosomal dominant inheritance. A heterozygous single base pair transition (CTG to CCG, leucine to proline) was detected in codon 518 of the betaig-h3 gene in the three affected members, and not in the unaffected member. No mutation was found in codon 124. Amyloid deposits were observed between the collagen bundles of the corneal stroma and were seen to extend deep into the stroma. CONCLUSION: The Leu518Pro mutated betaig-h3 forms amyloidogeneic intermediates which precipitate in the cornea and gives rise to a clinical appearance of LCDI. PMID- 10837381 TI - Glaucoma surgery with or without adjunctive antiproliferatives in normal tension glaucoma: 1 intraocular pressure control and complications. AB - BACKGROUND: Reduction of intraocular pressure (IOP) by 20-30% with glaucoma drainage surgery slows disease progression in normal tension glaucoma (NTG). It is not clear whether adjunctive antiproliferative agents are necessary or safe in eyes at low risk for scarring. METHOD: 86 eyes of 73 white NTG patients who had undergone a primary guarded fistulising procedure were reviewed. 25 eyes had no antiproliferatives, 36 had peroperative 5-fluorouracil (5-FU) and 25 had peroperative mitomycin C (MMC). Their postoperative IOPs, complications, and changes in visual acuity were recorded. RESULTS: Eyes that had no adjunctive antiproliferative less commonly maintained a 20-30% reduction in IOP (47.4% at 2 years) compared with either the 5-FU group (69.4%at 2 years, p=0.01) or the MMC group (64.9% at 2 years, p=0.04). Eyes that had adjunctive MMC more often had late hypotony (28%, p=0.02) and late bleb leak (12%, p<0.001). Eyes that had adjunctive MMC also more often had a two lines loss of Snellen visual acuity (39.8% by 2 years) compared with those that had adjunctive 5-FU (14.7% by 2 years), p=0.06. CONCLUSION: For NTG patients at low risk of scarring trabeculectomy with adjunctive peroperative 5-FU should maintain a suitable target IOP without the additional sight threatening complications seen with adjunctive MMC. PMID- 10837382 TI - Coagulation pathways and diabetic retinopathy: abnormal modulation in a selected group of insulin dependent diabetic patients. AB - AIMS: To investigate whether diabetic retinopathy (DR), already associated with microvascular alterations, ischaemia, and endothelial dysfunction, was also characterised by abnormal modulation of coagulation pathways. METHODS: Plasma samples, collected from 67 type 1 diabetics comparable for age, duration of disease (DD), and metabolic control (MC), were processed for prothrombin degradation products (F1+2) and factor VII coagulant activity (FVII:c). 50 normal subjects served as a control group. The ETDRS-Airlie House Classification of DR was used. RESULTS: A significant correlation between FVII:c and F1+2 plasma concentrations was observed (p <0.05). FVII:c (p <0.005) and F1+2 (p <0.0001) levels were higher in diabetics than in controls, especially in patients with proliferative DR (FVII:c p <0.0001; F1+2 p<0.005). However, cases without retinal lesions and healthy subjects did not differ significantly (FVII:c and F1+2 p >0.05). CONCLUSIONS: These findings pointed out the presence of a hypercoagulable state associated with endothelial dysfunction in patients with insulin dependent diabetes mellitus (IDDM), demonstrated both by increased FVII:c and F1+2 plasma levels. Moreover, the observation of different DR related degrees of procoagulant activity, despite comparable DD and MC, strengthens the hypothesis of multiple risk factors in the pathogenesis of DR. PMID- 10837383 TI - Intraventricular haemorrhage and stage 3 retinopathy of prematurity. AB - BACKGROUND/AIMS: A recent report has highlighted the decreasing prevalence in recent years of severe intraventricular haemorrhage (IVH) in very low birthweight (VLBW) infants (<1500 g). This study attempted to identify the severity of the grade of IVH in infants with stage 3 retinopathy of prematurity (ROP), and to re examine the association between threshold ROP and IVH. METHODS: This was a retrospective study carried out over 3 years, between December 1995 and December 1998 of neonates admitted to a single neonatal intensive care unit. 28 infants with stage 3 ROP were identified from the ROP screening database. Cranial ultrasound scans were available on 24 of these infants. The scans were reviewed and the severity of IVH was graded from grade 1 to grade 4. The birth weight, sex, ethnic origin, and gestational age of the babies were recorded. The number of infants progressing to threshold disease and the treatment provided was documented. RESULTS: The 24 infants had a median gestational age of 26 weeks (range 24-28 weeks) and a median birth weight of 762.5 g (range 540-1010 g). 17 infants were treated for threshold disease. 13 infants (54.2%) had IVH, of these eight (61.5%) had grade 1, two (15.4%) had grade 2, one (7.7%) had grade 3, and two (15.4%) had grade 4. 12 of the 13 infants (92.3%) with IVH had treatment with laser or cryotherapy for ROP compared with five of the remaining 11 infants (p = 0.023, Fisher's exact test). These data provide little evidence of any association between IVH and each of ethnic origin (p = 0.856), sex (p = 1), birth weight, or gestational age (p = 0.56 and p = 0.06 respectively) in infants with stage 3 ROP. CONCLUSIONS: These data provide strong evidence (p = 0. 023) of an association between the presence of IVH and treatment of threshold ROP. Although the numbers in this study are small the majority of infants with stage 3 ROP had grade 1 IVH, which heralds a more favourable neurological outcome. An association between the severity of ROP and severity of IVH was not demonstrated. With improvements in neonatal care and a reduction in the prevalence of severe IVH, there appears to be a weakening of the previously reported association between severe IVH and severe ROP. However, the presence of even a minor grade of IVH may be a significant risk factor for threshold ROP once stage 3 disease is encountered. PMID- 10837384 TI - Reversal of dysthyroid optic neuropathy following orbital fat decompression. AB - AIMS: To document the successful treatment of five patients with dysthyroid optic neuropathy by orbital fat decompression instead of orbital bone decompression after failed medical therapy. METHODS: Eight orbits of five patients with dysthyroid optic neuropathy were selected for orbital fat decompression as an alternative to bone removal decompression. Treatment with systemic corticosteroids and/or orbital radiotherapy was either unsuccessful or contraindicated in each case. All patients satisfied clinical indications for orbital bone decompression to reverse the optic neuropathy. High resolution computerised tomographic (CT) scans were performed in all cases and in each case showed signs of enlargement of the orbital fat compartment. As an alternative to bone decompression, orbital fat decompression was performed on all eight orbits. RESULTS: Orbital fat decompression was performed on five patients (eight orbits) with optic neuropathy. Optic neuropathy was reversed in all cases. There were no cases of postoperative diplopia, enophthalmos, globe ptosis, or anaesthesia. All patients were followed for a minimum of 1 year. CONCLUSIONS: In a subset of patients with an enlarged orbital fat compartment and in whom extraocular muscle enlargement is not the solitary cause of optic neuropathy, fat decompression is a surgical alternative to bony decompression. PMID- 10837385 TI - Role of reinsertion of the lower eyelid retractor on involutional entropion. AB - AIMS: To verify and evaluate the effect of reinsertion of the lower eyelid retractor aponeurosis to correct involutional entropion. METHODS: The involutional entropion is one affection that occurs mainly in the lower eyelid of patients over 60 years old. The surgical techniques proposed to correct this condition are based on correction of horizontal laxity-the preseptal orbicularis muscle overrides the pretarsal muscle, and the reinsertion of the lower eyelid retractor aponeurosis. 30 patients clinically diagnosed with involutional entropion and randomly selected underwent reinsertion of the lower eyelid retractor aponeurosis to the tarsal plate, without horizontal shortening or resection of the skin or orbicularis muscle. RESULTS: Good anatomical and functional correction was achieved in 96.6% of the patients and no recurrence was observed on 29 month follow up examination. The surgical result was very satisfactory. CONCLUSIONS: It was concluded that this procedure is effective and has low recurrence rate, showing the important role of the reinsertion of the lower eyelid retractor aponeurosis in this surgical correction. PMID- 10837386 TI - Treatment of recent onset central retinal vein occlusion with intravitreal tissue plasminogen activator: a pilot study. AB - AIMS: To study the effects of intravitreal tissue plasminogen activator (tPA) in recent onset central retinal vein occlusion (CRVO). METHODS: 15 patients with recent onset CRVO (from 1-21 days' duration, mean 8 days) were given 75-100 microg of tPA intravitreally associate with low dose low molecular weight heparin. CRVO was perfused in nine patients and with mild ischaemia not exceeding 100 disc diameters in six patients. Follow up ranged from 5 to 21 months for 14 patients (mean 8 months). Visual acuity measurement, macular threshold (Humphrey perimeter), fluorescein angiography with the scanning laser ophthalmoscope with special emphasis on retinal circulation times, and retinal perfusion were performed at days 0, 1, and 8 and months 1, 3, and 6. RESULTS: Visual acuity was significantly improved on the first day after treatment in only one eye, and decreased transiently in six eyes (40%). Retinal blood velocity was not significantly modified by tPA injection. Retinal ischaemia developed in six eyes (43%), leading to panretinal photocoagulation in five eyes including one with rubeosis iridis. At the end of follow up, visual acuity had improved to 20/30 or better in five eyes (36%), including two with complete recovery; visual acuity was worse than 20/200 in three eyes (28%). No complication of tPA injection was observed. CONCLUSION: Intravitreal tPA treatment for CRVO appears to be simple and safe, but did not significantly modify the course of the occlusion in our patients immediately after treatment. Final visual outcome did not differ significantly from that observed in the natural course of the disease, but final visual acuity seemed to be slightly better. A randomised study is required to determine if intravitreal tPA actually improves visual outcome in CRVO. PMID- 10837387 TI - Management of orbital lymphangioma using intralesional injection of OK-432. AB - AIM: To treat orbital lymphangioma with an intralesional injection of OK-432 (group A Streptococcus pyogenes of human origin). METHOD: A 14 year old boy had a right orbital cystic lymphangioma. The visual acuity in the eye was 20/28. In an initial treatment, 0.02 mg of OK-432, was injected into the tumour after aspiration of the fluid contents, but no effect was seen. The second treatment was performed with 0.04 mg of OK-432. RESULT: 4 months later, the lesion had totally shrunk to fibrous tissue. The side effects were fever, a local inflammatory reaction lasting 3 days, and increased intraocular pressure, which was managed by draining the fluid contents. Visual acuity improved to 20/15, and the visual field defect and restriction of eye movement seen before treatment disappeared. No recurrence was noted 1 year after treatment. CONCLUSION: An intralesional injection of OK-432 shrunk the lymphangioma without functional disturbance and scar in the facial skin. OK-432 may be useful for orbital lymphangioma, but further studies are still warranted to determine efficacy, complications, and the optimal dose for safe treatment. PMID- 10837388 TI - Comparative study of intraoperative mitomycin C and beta irradiation in pterygium surgery. AB - AIMS: To compare the rate of recurrence and complication after surgery for primary pterygium performed by one surgeon using either intraoperative mitomycin C or beta irradiation. METHODS: A retrospective study was performed of 164 eyes in 164 patients who had undergone primary pterygium surgery. After the pterygium was excised, the bare sclera was covered by sliding adjacent superior conjunctiva. 103 eyes received intraoperative mitomycin C (0.04%, 150 seconds) and 61 eyes beta irradiation (total dose 21.6 Gy). The mean follow up period was 20.2 (SD 17.9) months (range 1-66 months). Recurrence was defined as the postoperative regrowth of fibrovascular tissue crossing the corneoscleral limbus. RESULTS: The recurrence rate after mitomycin C and beta irradiation was 8.74% and 23.0% of eyes, respectively, after mean follow up of 17.9 and 31.2 months, respectively. The Kaplan-Meier survival analysis revealed a significantly better outcome for those who had intraoperative mitomycin C (Mantel-Cox log rank analysis, p=0.031). The mean interval to recurrence was not significantly different between the two groups. During the follow up, none of the patients showed side effects or reactions related to mitomycin C or beta irradiation. CONCLUSIONS: The intraoperative administration of 0.04% mitomycin C is more effective than beta irradiation as an adjunctive treatment for pterygium surgery in the patient population examined in this study. PMID- 10837389 TI - Contrast and glare sensitivity in epilepsy patients treated with vigabatrin or carbamazepine monotherapy compared with healthy volunteers. AB - BACKGROUND/AIM: Many antiepileptic drugs have influence on visual functions. The aim of this study was to investigate possible changes in contrast sensitivity, macular photostress, and brightness acuity (glare) tests in patients with epilepsy undergoing vigabatrin (VGB) or carbamazepine (CBZ) monotherapy compared with healthy volunteers. METHODS: 32 patients undergoing VGB therapy, 18 patients undergoing CBZ therapy, and 35 healthy volunteers were asked to participate in an ophthalmological examination. In the previous study, visual field constrictions were reported in 40% of the patients treated with VGB monotherapy. In the present study, these VGB and CBZ monotherapy patients were examined for photopic contrast sensitivity with the Pelli-Robson letter chart and brightness acuity and macular photostress with the Mentor BAT brightness acuity tester. RESULTS: Contrast sensitivity with the Pelli-Robson letter chart showed no difference between these groups and normal subjects (ANOVA: p= 0.534 in the right eye, p= 0.692 in the left eye) but the VGB therapy patients showed a positive correlation between the contrast sensitivity values and the extents of the visual fields in linear regression (R = 0.498, p = 0.05 in the right eye, R = 0.476, p = 0. 06 in the left eye). Macular photostress and glare tests were equal in both groups and did not differ from normal values. CONCLUSION: The results of this study indicate that carbamazepine therapy has no effect on contrast sensitivity. Vigabatrin seems to impair contrast sensitivity in those patients who have concentrically constricted in their visual fields. Neither GBZ nor VGB affect glare sensitivity. PMID- 10837390 TI - Mine blast injuries: ocular and social aspects. AB - BACKGROUND/AIMS: Landmines have long been used in conventional warfare. These are antipersonnel mines which continue to injure people long after a ceasefire without differentiating between friend or foe, soldier or civilian, women or children. This study focuses on Afghan non-combatants engaged in mine clearing operations in Afghanistan in the aftermath of the Russo-Afghan war. The patterns and types of injuries seen are described and experiences in their management, ways, and means to prevent them, and recommendations for the rehabilitation of the affected individuals are given. METHODS: It is a retrospective and analytical study of 84 patients aged 19-56 years who sustained mine blast injuries during mine clearing operations in Afghanistan from November 1992 to January 1996. The study was carried out at a military hospital with tertiary care facilities. The patients were divided into three groups on the basis of their injuries. Group 1 required only general surgical attention, group 2 sustained only ocular injuries, while group 3 had combined ocular and general injuries. Patients in groups 2 and 3 were treated in two phases. The first phase aimed at immediate restoration of the anatomy, while restoration of function wherever possible was done in subsequent surgical procedures in the second phase. RESULTS: It was observed that 51 out of 84 patients (60.7%) had sustained ocular trauma of a variable degree as a result of the blasts. The mean age of the victims was 29 years and they were all male. A total of 91 eyes of 51 patients (89.2%) had been damaged. Bilaterality of damage was seen in 40 (78.4%) patients. Most, 34 (37.3%), eyes became totally blind (NPL). Only a few escaped with injury mild enough not to impair vision. Foreign bodies, small and multiple, were found in the majority of eyes; most, however, were found in the anterior segment, and posterior segment injuries were proportionally less. CONCLUSIONS: The prevalence of blindness caused by mine blast injuries is quite high. The resulting psychosocial trauma to the patients and their families is tremendous and has not been adequately highlighted. These injuries are a great drain on the country's resources. Enforcement of preventive measures and the use of protective gear and sophisticated equipment by the mine clearing personnel would prove to be far more economical in terms of human life as well as medical and economic resources. There is also need for greater attention towards the establishment of support groups and rehabilitation programmes for these individuals. PMID- 10837391 TI - Basal nitric oxide production is enhanced by hydraulic pressure in cultured human trabecular cells. AB - BACKGROUND/AIMS: Nitric oxide donors reduce intraocular pressure. Human trabecular cells in culture were examined for their nitric oxide production in response to hydraulic pressure. METHODS: Human trabecular cells were cultured from trabeculum tissue fragments excised during trabeculectomy and exposed to hydraulic pressure change in a culture flask connected to a glass syringe. The pressure was exerted by automatic infusion of the piston of the syringe and monitored by a pressure gauge. The intracellular nitric oxide level was measured in real time with a nitric oxide binding fluorescent dye, diaminofluorescein-2. RESULTS: Intracellular nitric oxide levels in cultured trabecular cells showed spontaneous fluctuation during 400 seconds of observation. Peak levels of intracellular nitric oxide were significantly higher at hydraulic pressure of 30, 40, and 50 mm Hg, compared with 0 and 25 mm Hg (p<0.0001, one way ANOVA, and p<0.05, Tukey-Kramer test). The fluctuation was completely abolished by the presence of N-methyl-L-arginine (L-NMMA), a nitric oxide synthase inhibitor. The cultured trabecular cells were shown by immunohistochemistry to express brain nitric oxide synthase (bNOS). CONCLUSION: Higher levels of hydraulic pressure enhanced basal production of nitric oxide in human trabecular cells. Nitric oxide would be a physiological mediator in the regulation of intraocular pressure. PMID- 10837392 TI - Macrophage migration inhibitory factor levels in the vitreous of patients with proliferative diabetic retinopathy. AB - AIMS: To assess the potential role of macrophage migration inhibitory factor (MIF) in the pathogenesis of proliferative diabetic retinopathy (PDR). METHODS: MIF levels were assayed in the vitreous and paired serum samples of 73 consecutive patients with PDR (32 eyes) and macular hole or idiopathic epiretinal membrane (controls, 41 eyes). An enzyme linked immunosorbent assay technique was used to determine the concentrations of MIF. RESULTS: The median vitreous level of MIF was 11.93 ng/ml (range 4.16-103.85) in the patients with PDR, and 1.79 ng/ml (undetectable-8.93) in the controls. Vitreous levels in eyes with PDR were significantly greater than those in the controls (p<0.0001). Vitreous levels were significantly higher than serum levels in eyes with PDR (p=0.0026). MIF levels were significantly higher in the vitreous of PDR patients with severe fibrous proliferation than in those with slight proliferation (p<0.05). CONCLUSION: The results indicate increased levels of MIF in the vitreous of patients with PDR and a significant association between MIF levels and grades of fibrous proliferation, suggesting the possibility that MIF may play a part in the development of the proliferative phase of PDR. PMID- 10837393 TI - Novel materials to enhance keratoprosthesis integration. AB - BACKGROUND: The successful integration of keratoprostheses (KPros) within the cornea depends in part on peripheral host keratocyte adhesion to anchor the implant in place and prevent epithelial downgrowth. The following study incorporated different acrylate co-monomers with poly(hydroxyethyl methacrylate) (p(HEMA)) and measured the suitability of these materials as potential skirt materials in terms of their ability to enhance keratocyte adhesion to p(HEMA). METHODS: p(HEMA) hydrogels incorporating varying amounts of the acrylate co monomers methacrylic acid (MA), 2-(dimethylamino)ethyl methacrylate (DEM), or phenoxyethyl methacrylate (PEM) were formed by free radical polymerisation. Keratocytes were seeded onto discs of each material and incubated at 37 degrees C for 72 hours. Assays for viable cell adhesion were carried out. A viability/cytotoxicity assay using solutions of calcein-AM (0.5 mM) and ethidium homodimer-1 (EthD-1) (0.5 microM) were used to measure viable and non-viable cell adhesion, respectively. An ATP assay was also used to quantify cell adhesion in terms of the amount of ATP present following lysis of adherent cells. RESULTS: The viability/cytotoxicity assays indicated that the incorporation of 15 mol% of the co-monomer PEM or of 20 mol% DEM increased cell adhesion to p(HEMA) by at least four times. The ATP assays confirmed the results for PEM but absorption of ATP to the DEM containing hydrogel indicated that the assay was not a suitable measure of cell adhesion to this material. CONCLUSIONS: The properties of p(HEMA) may be moderated to enhance keratocyte adhesion by the incorporation of PEM or DEM suggesting that these may be suitable materials for use in the further development of a novel KPro skirt material. PMID- 10837394 TI - Automated, real time extraction of fundus images from slit lamp fundus biomicroscope video image sequences. AB - AIMS: Slit lamp fundus biomicroscopy allows for high magnification, stereoscopic diagnosis, and treatment of macular diseases. Variable contrast, narrow field of view, and specular reflections arising from the cornea, sclera, and examining lens reduce image quality; these images are of limited clinical utility for diagnosis, treatment planning, and photodocumentation when compared with fundus camera images. Algorithms are being developed to segment fundus imagery from slit lamp biomicroscopic video image sequences in order to improve clinical utility. METHODS: Video fundus image sequences of human volunteers were acquired with a video equipped, Nikon NS-1V slit lamp biomicroscope. Custom developed software identified specular reflections based on brightness and colour content, and extracted the illuminated fundus image based on colour image analysis and size constraints. RESULTS: In five subjects with variable image quality, the approach allowed for automatic, robust, accurate extraction of that portion of the video image corresponding to the illuminated portion of the fundus. Non-real time analysis allowed for fundus image segmentation for each frame of the image sequence. In real time, segmentation occurs at 2 Hz, and improvements are being implemented for video rate performance. CONCLUSIONS: Computer vision algorithms allow for real time extraction of fundus imagery from marginal quality, slit lamp fundus biomicroscope image sequences. PMID- 10837395 TI - Conjunctival epithelium expression of HLA-DR in glaucoma patients and its influence on the outcome of filtration surgery. AB - AIMS: To analyse the expression of HLA-DR on conjunctival epithelial cells in patients with glaucoma taking topical antiglaucoma therapy. METHODS: 10 patients taking no topical medication and 30 patients with uncontrolled glaucoma taking medical therapy participated in the study. The specimens were obtained by impression cytology preoperatively, 3 months, and 6 months after filtration surgery. The expression of HLA-DR on T lymphocytes and epithelial cells was analysed by flow cytometry. RESULTS: A significant increase in HLA-DR on epithelial cells was found preoperatively in patients with glaucoma. A significant increased expression of HLA-DR on epithelial cells was detected 3 months and 6 months after surgery. CONCLUSIONS: The increased expression of HLA DR on conjunctival epithelial cells still present 6 months after surgery indicates the increased ability of epithelial cells to induce immune inflammation with subsequent fibrosis. PMID- 10837396 TI - Influence of some operational variables on the radial keratotomy operation. AB - AIMS: To assess the contribution of the four major operational variables in the radial keratotomy operation (RK) to the correction of myopic eyes. To study the deformation of the cornea after the operation and provide some valuable references for clinical practice. METHODS: The expression of the correction ratio has been deduced, which can be directly represent as the deformation ratio of the corneal radius after the operation. This allows for the numerical simulation of the RK operation. On the basis of the known biomechanical property of the cornea, by means of the finite element method, a series of computerised geometric and biomechanical models for the simulation of refractive surgery have been established. Some operational variables in the RK operation were analysed, and their influence on the operational outcome has been computed and systematically analysed. RESULTS AND CONCLUSION: All of the four variables have a positive effect on the operational outcome. The greater the four variables are the more correction effects of myopia can be obtained. Because the four variables are interinfluential, the operational variables can be optimised to get the best result in order to meet patients' differing requirements. PMID- 10837398 TI - Prophylactic scleral buckle for prevention of retinal detachment following vitrectomy for macular hole. PMID- 10837399 TI - Bilateral optic disc oedema associated with latanoprost. PMID- 10837397 TI - Matrix metalloproteinase biology applied to vitreoretinal disorders. PMID- 10837400 TI - Membrane formation in the chamber angle after failure of argon laser trabeculoplasty. PMID- 10837401 TI - CpG motifs induce Langerhans cell migration in vivo. AB - Cytosine-guanosine (CpG) oligonucleotide (CpG-oligo) sequences are immunostimulatory motifs that are present in bacterial DNA and their presence in plasmids might contribute to the immune response generated by DNA vaccination. The cell targets of CpG motifs in vivo have not been characterized yet. In this report we assessed the in vivo effects of CpG motifs on Langerhans cells (LC) migration. We showed that intradermal injection of 10 microg of CpG-containing oligonucleotides in mouse ear induced the local depletion of LC within 2 h of exposure as shown by CD11c and Ia immunohistological staining. To demonstrate that LC depletion was due to LC migration, CpG oligonucleotides were injected into the explants ex vivo, and the CD11c(+) cells emigrating from the cultured isolated skin within medium were evaluated by immunostaining and FACS analysis. Our findings demonstrate that CpG motifs induce LC/dendritic cell (DC) migration out of the skin. To assess whether CpG motifs may act directly on LC/DC to induce their emigration we next analyzed the effects of CpG motifs in vitro on the expression of adhesion molecules involved in LC/DC migration. The results of these experiments show that alpha(6) integrins, E-Cadherin, ICAM-1, CD11b and CD11c were differentially regulated upon CpG-oligo treatment of immortalized DC. CpG treatment (10 microg/ml for 8 h) resulted in a 100% increase in ICAM-1 staining intensity, a 50% decrease in E-Cadherin staining and a 25% decrease in alpha(6) integrins staining, while no changes in the levels of CD11b and CD11c expression were recorded. Changes in adhesion molecule expression were mirrored by concomitant changes in the cell morphology that included cell depolarization, the appearance of filopods and loss of adherence. This study provides the first in vivo evidence that CpG motifs signal the migration of LC from the epidermis. PMID- 10837402 TI - An example of immunodominance: engagement of synonymous TCR by invariant CDR3 beta. AB - The structural basis of the T cell response against immunodominant tetanus toxin (TT)-derived peptides was investigated using TT-specific T cell clones raised from a DRB1*0301 homozygous donor. Three peptides forming T cell epitopes were identified, including one, TT(1272-1284), that stimulated four different TT specific T cell clones. TCR sequence analysis revealed that these synonymous TCR shared only arginine at the third position of the CDR3 beta loop. This prominent residue may form a salt bridge with a corresponding aspartate at the relative position 8 (P8) of the antigenic peptide TT(1272-1284) as suggested from amino acid replacement analysis. A similar scenario was observed for a second TT epitope, TT(279-296), and its corresponding TCR. These examples show that immunodominance may result from a single strong amino acid interaction between TCR CDR3 beta loops here in contact with the C-terminus of the antigenic peptide. Such a dominant interaction could compensate for weaker contacts between other residues of the TCR and the antigenic peptide, and would allow the recognition of a single peptide-MHC complex by a broader synonymous TCR repertoire and could thus contribute to its immunodominance. PMID- 10837403 TI - The functional role of class II-associated invariant chain peptide (CLIP) in its ability to variably modulate immune responses. AB - During the process of class II MHC assembly and cell surface expression, the class II-associated invariant chain peptide (CLIP) is removed from the peptide binding groove of MHC, a task mediated by H-2M. This allows binding and presentation of peptide epitopes. We have previously shown that exogenously added CLIP interferes with this process and down-regulates the cell surface expression of class II molecules. In this study, we explored the effect of exogenously added CLIP on antigen-specific immune responses. In vivo studies with CLIP and various peptide and protein antigens with different affinities for I-A(d) molecules demonstrated that CLIP variably affects the T cell-mediated immune responses. Immunization with CLIP along with the antigen induced a shift from a T(h)1- to T(h)2-like response as determined by the cytokine profile and antibody isotype. These results suggest that the presence of exogenous CLIP can significantly influence the presentation of antigen by class II MHC molecules to CD4 T cells and thereby modulate immune responses. Exogenously added CLIP rapidly localized into the subcellular compartment of antigen-presenting cells where MHC class II molecules are present. We suggest that exogenous CLIP reduces the loading of peptides on the class II molecules, thus down-regulating MHC-peptide complexes on the cell surface. Alternatively, CLIP may bind to cell surface class II molecules and this complex is rapidly internalized resulting in reduced cell surface MHC class II expression. The reduced level of MHC-peptide complexes favors the activation of T(h)2 cells over T(h)1 cells. These results have implications in the regulation of immune responses, particularly the prevention of certain autoimmune diseases where T(h)1-type responses are pathogenic and T(h)2-type responses are protective. PMID- 10837404 TI - The influence of CD40-CD154 interactions on the expressed human V(H) repertoire: analysis of V(H) genes expressed by individual B cells of a patient with X-linked hyper-IgM syndrome. AB - Analysis of the V(H)DJ(H) repertoire of peripheral blood IgM(+) B cells from a patient with X-linked hyper-IgM syndrome (X-HIgM) was undertaken to determine whether the distribution of V(H) families in the productive repertoire might be regulated by in vivo CD40-CD154 interactions. The distribution of V(H) genes in the non-productive repertoire of IgM(+) B cells was comparable in X-HIgM and normals. Unlike the normal productive V(H) repertoire, however, in the X-HIgM patient the V(H)4 family was found at almost the same frequency as the V(H)3 family. This reflected a diminution in the positive selection of the V(H)3 family observed in normals and the imposition of positive selection of the V(H)4 family in the X-HIgM patient. Unique among the V(H)3 genes, V(H)3-23/DP-47 was positively selected in both normals and the X-HIgM patient. No major differences in the usage of J(H) or D segments or the complementarity-determining region (CDR) 3 were noted, although the foreshortening of the CDR3 noted in the mutated V(H) rearrangements of normals was absent in the X-HIgM patient. Finally, a minor degree of somatic hypermutation was noted in the X-HIgM patient. These results have suggested that specific influences on the composition of the V(H) repertoire in normals require CD40-CD154 interactions. PMID- 10837405 TI - Mutations in specific I-A(k) alpha(2) and beta(2) domain residues affect surface expression. AB - A previous investigation demonstrated that several mutations in class II dimer-of dimers contact residues interfere with antigen presentation by transfectants but not with plasma membrane expression of the mutant class II. In the present study we examined other class II mutations in this region that did inhibit plasma membrane expression of mutant class II molecules. Molecules containing both mutations H alpha 181D in the alpha(2) domain and E beta 170K in the beta(2) domain exhibited low plasma membrane expression, but molecules with only one of these mutations were expressed normally. The mutant class II molecules were transported to organelles that were accessible to a fluid-phase protein, hen egg lysozyme (HEL). Culture of transfectants with lysozyme enhanced the amount of class II compact dimer (alpha beta plus peptide; CD), and this was especially marked for the class II mutant H alpha 181D/E beta 170K and for other molecules possessing both mutations. Formation of class II CD was not paralleled by an increase in class II surface expression. Thus the joint mutation of H alpha 181 and E beta 170 has two effects. In the absence o high concentrations of exogenous peptide, it prevents efficient CD formation, possibly by affecting invariant chain (Ii) proteolysis and/or the stability of the class II after Ii/CLIP is removed. At high peptide concentrations supplied by exogenous HEL, the mutations allow CD formation, but not expression of class II on the plasma membrane. Molecular modeling of the possible interaction of class II and Ii suggests that the mutant amino acids H alpha 181D and E beta 170K, besides affecting the overall stability of class II, might also interact with Ii via two loops in class II's alpha(2) and beta(2) domains respectively. PMID- 10837406 TI - CD40 expressed on human melanoma cells mediates T cell co-stimulation and tumor cell growth. AB - CD40 is a 50 kDa molecule, a member of the tumor necrosis factor/nerve growth factor receptor family. It is expressed on B cells, monocytes, dendritic cells and various malignant cells. While the critical relevance of this molecule in T cell-dependent B cell activation is already established, the biological role of CD40-CD154 interaction in non-hematopoietic cells is still unknown. Here we show that CD40 is functionally expressed on human melanoma-derived cell lines. No correlation between surface CD40 expression and the origin of the cell line, primary versus metastatic, was observed. Melanoma cells were shown to be able to co-stimulate TCR-triggered human T cells; moreover, because they do not express CD80 or CD86 co-stimulatory structures, the involvement of additional pathways have to be postulated. We have identified CD40 as one of the molecules involved in melanoma cell-mediated co-stimulation of anti-CD3-triggered human CD4(+) T lymphocytes. In addition, a CD40-dependent pathway, able to enhance tumor cell proliferation at low serum concentrations, in vitro, has been shown to be functional in human melanoma cell lines. PMID- 10837407 TI - High frequency of circulating gamma delta T cells with dominance of the v(delta)1 subset in a healthy population. AB - TCR gamma delta(+) cells constitute <5% of all circulating T cells in healthy, adult Caucasians, and V(delta)1(+) cells constitute a minority of these cells. In contrast to TCR alpha beta(+) cells, their repertoire is selected extrathymically by environmental antigens. Although increased frequencies of V(delta)1(+) cells are found in several diseases, their function remains obscure. Here we show that the frequency of peripheral blood gamma delta T cells in healthy West Africans is about twice that of Caucasians, mainly due to a 5-fold increase in V(delta)1(+) cells, which is consequently the dominant subset. No age dependency of V(delta)1 frequencies was identified and the V(delta)1(+) cells in the African donors did not show preferential V(gamma) chain usage. Analysis of the CDR3 region size did not reveal any particular skewing of the V(delta)1 repertoire, although oligoclonality was more pronounced in adults compared to children. The proportions of CD8(+), CD38(+) and CD45RA(hi)CD45RO(-) cells within the V(delta)1(+) subset were higher in the African than in the European donors, without obvious differences in expression of activation markers. No significant correlations between levels of V(delta)1(+) cells and environmental antigens or immunological parameters were identified. Taken together, the evidence argues against a CDR3-restricted, antigen-driven expansion of V(delta)1(+) cells in the African study population. Our study shows that high frequencies of TCR gamma delta(+) cells with dominance of the V(delta)1(+) subset can occur at the population level in healthy people, raising questions about the physiological role of V(delta)1(+) T cells in the function and regulation of the immune system. PMID- 10837408 TI - The adjuvant monophosphoryl lipid A increases the function of antigen-presenting cells. AB - The induction of immune responses in vivo is typically performed with antigens administered in external adjuvants, like alum, complete Freund's adjuvant, LPS and, more recently, monophosphoryl lipid A (MPL). However, the role of the adjuvant is still poorly defined. The aim of this study was to test whether the MPL affects the function of antigen-presenting cells (APC) in vitro and in vivo. Antigen-pulsed APC [including macrophages, B cells and dendritic cells (DC)] were incubated or not with MPL, and their ability to sensitize naive T cells was tested in vitro and in vivo. The data show that MPL enhances the ability of macrophages and B cells to sensitize naive T cells, and confers to them the capacity to induce the development of T(h)1 and T(h)2. Administration of MPL i.v. in mice results in the redistribution of fully mature DC in the T cell area of the spleen. These observations suggest that MPL may induce an antigen-specific primary immune response by provoking the migration and maturation of DC that are the physiological adjuvant of the immune system. PMID- 10837409 TI - Impaired Ca/calcineurin pathway in in vivo anergized CD4 T cells. AB - Clonal anergy is one of the mechanisms that may account for self tolerance induced in T cells in the periphery. In this study we used the well-documented system of in vivo administration of a superantigen, staphylococcal enterotoxin B (SEB), to induce a state of hyporesponsiveness (anergy) in murine peripheral T cells to decipher the intracellular biochemical basis for this process. The TCR induced Ca response of in vitro activated T cells was found to be impaired with significant defects in the phosphorylation of phospholipase C-gamma 1. Experiments with calcium ionophore and newly established transgenic mouse lines that express an active form of calcineurin suggested that in vivo SEB-induced anergy is established and/or maintained by a selective impairment in the TCR induced activation of the Ca/calcineurin pathway. PMID- 10837410 TI - Dendritic cells at a DNA vaccination site express the encoded influenza nucleoprotein and prime MHC class I-restricted cytolytic lymphocytes upon adoptive transfer. AB - Intradermal inoculation of plasmids expressing antigens that contain MHC class I restricted epitopes leads to the induction of specific CD8(+) cytotoxic T lymphocytes (CTL). The role of in situ transfected antigen-presenting cells (APC) in the priming of specific CTL subsequent to intradermal DNA immunization was investigated using a plasmid (NPV1) expressing the nucleoprotein (NP) of influenza virus that contains a nuclear targeting signal and a dominant class I/K(d)-restricted epitope. Inoculation of NPV1 leads to in situ transfection of MHC class II(+) and class II(-) cells, as revealed by the nuclear localization of NP. Between 2 and 3% of MHC class II(+) and class II(-) cells with the ability to migrate out of the epidermis expressed NP. Upon adoptive transfer into naive recipients, class II(+) migratory cells recovered from the area inoculated with NP-expressing plasmid were significantly superior regarding the ability to prime virus-specific CTL as compared to MHC class II(-) cells. Together, these results are consistent with the role of local dendritic cells loaded with antigen in the priming of CTL by intradermal DNA immunization. PMID- 10837411 TI - Enhanced surface TCR replenishment mediated by CD28 leads to greater TCR engagement during primary stimulation. AB - When T cells are stimulated with high concentrations of strong TCR agonist, engaged TCR are internalized and degraded, resulting in greatly reduced surface TCR levels for up to several days post-stimulation. It has been noted that surface TCR levels rise subsequently, even in the presence of continuing stimulation, but the role of CD28 co-stimulation in surface TCR replenishment has not been investigated. Here, we have examined the return of surface TCR following activation, the availability of these TCR for antigenic engagement and the role of CD28 in that process. We report that within 24 h of stimulation, the level of surface TCR expression becomes dependent on the degree of CD28 signaling provided during T cell activation. In addition, when cells are removed from stimulus after 24 h, surface TCR expression recovers to a stable level which exceeds that of unstimulated cells and is proportional to the degree of CD28 co-stimulation. TCR that replenish the plasma membrane during T cell activation can be down-regulated by receptor occupancy with the same efficiency as TCR on freshly stimulated cells. Thus, as a result of enhanced surface TCR replenishment, CD28-co stimulated cells can engage and down-regulate more TCR than co-stimulation deprived cells in the face of ongoing stimulation. Furthermore, engagement of newly expressed TCR on activated T cells re-induces CD69, suggesting participation of these replenishing TCR in continued T cell signaling. These data identify the augmentation of surface TCR replenishment during activation as a novel mechanism that likely contributes to the enhanced antigenic sensitivity of CD28-co-stimulated T cells. PMID- 10837412 TI - Self-MHC class Ia (RT1-A(n)) protects cells co-expressing the activatory allogeneic MHC class Ib molecule (RT1-E(u)) from NK lysis. AB - We have previously shown activation of NK cells via recognition of an allogeneic, non-classical MHC class I molecule, RT1-E(u). In this study we investigated whether a self-MHC class I molecule could protect the allogeneic targets from being recognized and killed by the alloreactive NK (allo NK) cells. NK cells from BN (RT1 n) rats, primed in vivo by immunization with RT1(u)-expressing cells, manifested cytolytic activity against RT1(u)- as well as RT1(u/lv1)-expressing targets, but not against RT1(u/n)-expressing targets. The absence of cytolytic activity against semiallogeneic targets, i.e. targets expressing self-allotypes, was also valid for allo NK cells from alloimmunized F344 (RT1 (lv1)) rats. To analyze the ability of a distinct MHC class I molecule to protect target cells from NK lysis, Rat2 cells transfected with the activating allogeneic MHC class Ib, RT1-E(u) molecule were also transfected with the self-MHC class Ia, RT1-A1(n) molecule. The allo NK cells from BN rats immunized with RT1(u)-expressing cells were cytolytic against Rat2 transfected with the RT1-E(u) molecule. However, the allo NK cells manifested no cytolytic activity against double-transfected Rat2 cells, expressing the RT1-E(u) as well as the RT1-A1(n) molecule. We conclude that expression of a self-MHC class Ia (RT1-A) molecule protects targets from allo NK killing. Furthermore, the NK inhibition via recognition of the self-MHC class Ia molecule dominates over the activation via recognition of the allogeneic MHC class Ib molecule, RT1-E. PMID- 10837413 TI - Regulation of TCR-induced IFN-gamma release from islet-reactive non-obese diabetic CD8(+) T cells by prostaglandin E(2) receptor signaling. AB - Prostaglandins (PG) are released during tissue injury and inflammation, and inhibit immune responses at many points. PG may be one of several factors that protect not only against injury-induced, but also spontaneous, organ-specific autoimmune disease. Here we show that the production of PGE(2), normally produced at a very low rate in islets of Langerhans, is significantly increased in inflamed islets of non-obese diabetic (NOD) mice. We investigated a possible role of PGE(2) in controlling TCR-dependent release of IFN-gamma from islet-reactive NOD CD8(+) T cells. PGE(2) inhibited anti-TCR antibody-triggered release of IFN gamma from CD8(+) T cell clone 8D8 and from polyclonal cytotoxic T lymphocytes (CTL). Using receptor subtype selective agonists, we present evidence that the effect of PGE(2) is mediated by EP(2) and EP(4) receptors, both of which are coupled to an increase in intracellular cAMP production. The cAMP analogs 8-Br cAMP and Sp-cAMPS mimic the effect of EP(2)/EP(4) receptor agonists, inhibiting TCR-triggered IFN-gamma release from NOD CD8(+) T cells in a dose-dependent manner. The inhibitory effect of PGE(2) was largely reversed by IL-2 added at the time of culture initiation and decreased with increasing strength of stimulation through the TCR. Resting CTL were more sensitive to PGE(2) than recently expanded CTL and NOD CD8(+) T cells remained insensitive to PGE(2) for a longer time than BALB/c cells. Our study suggests that PGE(2) may be part of a regulatory network that controls local activation of T cells and may play a role in the balance between the development of islet autoimmunity or maintenance of tolerance. PMID- 10837414 TI - Impairment of B lymphopoiesis in precocious aging (klotho) mice. AB - Inactivation of the klotho gene in mice results in multiple disorders that resemble human aging after 3 weeks of age. Because hematopoiesis, especially B lymphopoiesis, is affected in humans and mice by aging, we analyzed the hematopoietic state in homozygous klotho (kl/kl) mice. The kl/kl mice showed thymic atrophy and a reduced number of splenocytes. These mice had almost the normal number of myeloid cells, erythroid cells, IL-3-responsive myeloid precursors and colony forming units in spleen (CFU-S) in bone marrow (BM), but had a substantially decreased number of B cells in BM and peripheral blood as compared with wild-type mice. IL-7-responsive B cell precursors and all of the maturation stages of B cells in BM were also reduced. However, the function of hematopoietic stem cells including their capacity of B lymphopoiesis in vivo and in vitro was normal. Early B cell development was also normal in neonates and young kl/kl mice until 2 weeks old without aging phenotypes. RT-PCR analysis revealed that the level of IL-7 gene expression was significantly reduced in freshly isolated kl/kl BM cells. However, injection of IL-7 in kl/kl mice could not rescue the B lymphopenia. These findings indicate that Klotho protein may regulate B lymphopoiesis via its influence on the hematopoietic microenvironment. PMID- 10837415 TI - Positive and negative selection of antigen-specific B cells in transgenic mice expressing variant forms of the V(H)1 (T15) heavy chain. AB - Four variant forms of the V1 (T15-H chain) gene are synthesized in mice. Each V1 variant pairs with a distinct L chain to produce a binding site having specificity for phosphocholine (PC). Transgenic mice expressing variant forms of the V1 gene were analyzed to elucidate the factors driving B cell selection into the peripheral repertoire. In all four lines of H chain transgenic mice analyzed, transgene expression caused complete allelic exclusion of endogenous H chains in the bone marrow (BM), whereas most splenic B cells expressed endogenous H chains. The number of sIgM(+) BM B cells and their sIg receptor number was reduced compared to that of normal transgene-negative controls, suggesting that B cells expressing transgene-encoded H chains were being negatively selected in the BM. Mice expressing autoreactive forms of the V1 transgene with lower affinity for PC (M603H and M167H) exhibit positive selection of PC-specific B cells into the spleen, whereas mice expressing the higher affinity T15H variant exhibited elevated PC-specific B cells in the peritoneal cavity but few V(H)1(+) splenic B cells. These data suggest that the higher affinity T15-id(+) B cells preferentially survive in the peritoneal cavity. When these H chain transgenes were crossed into the mu MT knockout mouse in which surface expression of endogenous H chains is blocked, the percent of splenic V(H)1(+) PC-specific B cells increased up to 5-fold and T15-id(+) B cells were detectable in the spleen of T15H mice. This implies that T15-id(+) PC-specific B cells can be selected into the periphery, but they compete poorly with follicular B cells expressing endogenous Ig. PMID- 10837416 TI - In vitro and in vivo macrophage function can occur independently of SLP-76. AB - Expression of SH2 domain-containing leukocyte-specific phosphoprotein of 76 kDa (SLP-76), a hematopoietic cell-specific adapter protein, is required to couple Syk family tyrosine kinase activation to downstream mediators such as phospholipase C (PLC)-gamma following TCR, platelet collagen receptor and mast cell Fc epsilon R stimulation. In addition to T cells, mast cells and platelets, SLP-76 is expressed in monocytes and macrophages. To determine the role of SLP-76 in Fc gamma R-stimulated signaling pathways in macrophages, we examined cultured bone marrow-derived macrophages (BMM) from SLP-76(-/-) and wild-type mice. In this study, we show that Fc gamma R cross-linking rapidly induces tyrosine phosphorylation of SLP-76 in wild-type BMM. Surprisingly, however, BMM from SLP 76(-/-) mice activate ERK2 and phosphorylate PLC-gamma 2 following Fc gamma R ligation. Furthermore, SLP-76(-/-) BMM display normal Fc gamma R-dependent phagocytic function and reactive oxygen intermediate production. SLP-76(-/-) and SLP-76(+/+) BMM secrete comparable levels of IL-12 in response to lipopolysaccharide and IFN-gamma. To examine macrophage function in vivo, SLP-76( /-) mice were challenged i.v. with Listeria monocytogenes. SLP-76(-/-) mice survive and efficiently contain the acute phase of infection similar to wild-type mice but exhibit a stable chronic infection attributed to the lack of mature T cells. These data show that, although SLP-76 is required to couple Syk family PTK activity to downstream mediators and effector functions in Fc gamma R-induced pathways in some cell types, activation of Fc gamma R-dependent pathways occurs independently of SLP-76 in BM PMID- 10837417 TI - A new mechanism for IL-5-dependent helminth control: neutrophil accumulation and neutrophil-mediated worm encapsulation in murine filariasis are abolished in the absence of IL-5. AB - IL-5 production and eosinophilia are features of helminth infections, but results concerning the role of IL-5 and eosinophils (EP) in worm control are contradictory. We describe here a novel, IL-5-dependent mechanism of helminth control in vivo, using a fully permissive murine filariasis model, i.e. infection of BALB/c mice with Litomosoides sigmodontis. Worm control was exerted by the formation of inflammatory nodules around adult filariae which initially remained alive but were eventually killed within several weeks. The cell population essential for inflammatory nodule formation was found to be neutrophils (NP) but not EP. Neutralization of IL-5 led to a failure of both EP and NP accumulation at the site of infection (i.e. the thoracic cavity), resulting in cessation of inflammatory nodule formation around worms and in their survival. The role of NP in this process was confirmed by treatment of mice with anti-granulocyte colony stimulating factor (G-CSF) which also resulted in a lack of inflammatory nodule formation and worm killing albeit in the presence of EP. Since IL-5, due to the absence of IL-5 receptors on NP, does not act on these cells directly, it was investigated if anti-IL-5 altered the production of NP-chemotactic cytokines. In anti-IL-5-treated mice, cytokines known to promote NP accumulation like tumor necrosis factor-alpha, G-CSF and KC (IL-8) were found to be strongly reduced, while NP-deactivating cytokines like IL-10 were increased. In conclusion, IL-5 constitutes a cytokine essential for NP-mediated worm control in filarial infection. PMID- 10837418 TI - Relative contribution of NK and NKT cells to the anti-metastatic activities of IL 12. AB - Conventional T cells, NK cells and NKT cells have been implicated in the anti tumor activities induced by IL-12. Here we show that IL-12-induced immune responses are partially impaired in T and NKT cell-deficient RAG-2(-/-) mice, and in NKT cell-deficient CD1(-/-) mice. In response to a small dose (<1000 U) of IL 12, RAG-2(-/-) and CD1(-/-) mice demonstrated reduced cytotoxicity, serum IFN gamma elevation and anti-metastatic activities; in contrast, in response to a high dose (>2000 U) of IL-12, the IL-12-induced immune responses of RAG-2(-/-) and CD1(-/-) mice were indistinguishable from wild-type mice. The defective responses to low-dose IL-12 of RAG-2(-/-) mice were corrected by adoptive transfer of NKT cells but not NK cells. These findings indicate that both NK and NKT cells contribute to the anti-metastatic responses induced by IL-12, and that NKT cells are mostly responsible for the low-dose activities of this cytokine. PMID- 10837419 TI - Alterations in splenic architecture and the localization of anti-double-stranded DNA B cells in aged mice. AB - Aging is characterized by a decline in humoral immunity and a concommitant increased incidence of anti-DNA and other autoantibodies. To define how the regulation of autoreactive B cells is altered with age, we have used BALB/c mice with an Ig heavy H chain transgene to track the fate of anti-double-stranded (ds) DNA B cells in vivo. In young adult mice, anti-dsDNA B cells are developmentally arrested and excluded from the splenic B cell follicle, whereas in most aged mice they are mature and localize within the B cell follicle. Furthermore, we have detailed global changes in lymphoid architecture that accompany aging: CD4(+) T cells are found not only in the periarteriolar lymphoid sheath, but also in the B cell follicles. Strikingly, these disruptions are similar to those that precede serum anti-dsDNA antibody expression in autoimmune MRL-lpr/lpr mice. PMID- 10837420 TI - Membrane and transmembrane signaling in Herpesvirus saimiri-transformed human CD4(+) and CD8(+) T lymphocytes is ATM-independent. AB - In the genetic disorder ataxia telangiectasia (AT), humoral (B) and cellular (T) immunological abnormalities are frequently observed. As a consequence, AT patients are predisposed to life-threatening sinopulmonary infections. The pathogenic mechanisms remain unknown, but a role for ATM in signal transduction from membrane receptors has been proposed. We have explored the effects of a defective ATMgene on isolated human T-lineage cells from 13 AT patients with proven T cell dysfunction by transforming their CD4(+) and CD8(+) T lymphocytes with Herpesvirus saimiri, and analyzing their signaling behavior as compared to normal controls. Several functional parameters were assayed in response to both membrane (anti-CD3 and IL-2) and transmembrane (phorbol myristate acetate plus the calcium ionophore ionomycin) stimuli: (i) calcium mobilization, (ii) induction of activation molecules (CD25, CD40 ligand, CD69 and CD71), (iii) cytokine synthesis (IL-2 and tumor necrosis factor-alpha) and (iv) proliferation. All these early and late activation events were found to be normal in the transformed ATM-/-T cells, indicating that ATM is not necessary for their induction. As expected, ATM-/- transformed T cells showed an increased radiosensitivity by both radioresistant DNA synthesis and cell survival assays. In contrast to an earlier report testing transformed B lymphocytes, our results indicate that transformed mature peripheral T lymphocytes from AT patients do not have intrinsic immune function defects. Rather, the described T-lineage signaling impairments observed in patients may be secondary in vivo to extrinsic ATM dependent suppressive factors and/or to a developmental defect. These transformed T cells may help to understand the distinct biological role of ATM in different cell types and to develop rational therapies for the immunological dysfunction of AT patients. PMID- 10837421 TI - T(h)1 transmigration anergy: a new concept of endothelial cell-T cell regulatory interaction. AB - Stimulation of endothelial cells (EC) with IFN-gamma generates selective enhancement of T(h)1 cell transmigration and induction of MHC class II expression on EC. In the present study, we tested whether antigen presentation by EC could influence transmigrating T cells in an in vitro system. Bacterial antigen presentation by EC from primary culture and after cloning induced antigen specific anergy of transmigrating T(h)1 clone cells in a MHC class II-dependent manner as characterized by non-responsiveness to subsequent antigen presentation and inability to produce IL-2. This T cell transmigration anergy induced by EC was abrogated by anti-rat CD28 mAb, suggesting that lack of B7 co-stimulatory signals by EC might be related to the induction of anergy. While MHC class II expression on primary and cloned EC was observed after IFN-gamma stimulation, these cells never expressed B7. B7-1 gene-transfected endothelial clone cells (ECC/B7-1) were developed to elucidate the influence of B7 co-stimulation by EC. ECC/B7-1 induced proliferation of T(h)1 clone cells, whereas ECC did not induce proliferation in co-culture of T(h)1 clone cells and EC stimulated with IFN-gamma and antigen. In the transmigration assay, ECC/B7-1 did not induce transmigration anergy of T(h)1 clones or T(h)1 lines unless anti-rat B7-1 blocking mAb was added. Therefore, in rats, the T cell anergy induced during transmigration across a layer of EC seemed to be due to antigen presentation in the absence of B7 on the EC. We introduce the concept of transmigration anergy in this manuscript. Thus, EC can play a critical immune regulatory role in the context of antigen presentation by MHC class II to transmigrating T cells. PMID- 10837422 TI - Minimizing quinolone resistance: are the new agents more or less likely to cause resistance? PMID- 10837423 TI - Outpatient parenteral treatment of bacterial infections: the Italian model as an international trend? PMID- 10837424 TI - Management of herpes virus infections following transplantation. PMID- 10837425 TI - Invasive mycoses in organ transplant recipients: controversies in prophylaxis and management. PMID- 10837426 TI - Towards a common susceptibility testing method? PMID- 10837427 TI - Expression of resistance to tetracyclines in strains of methicillin-resistant Staphylococcus aureus. AB - A diverse collection of methicillin-resistant Staphylococcus aureus (MRSA) isolates resistant to tetracycline was screened by PCR for the presence of the resistance determinants tetK, tetL, tetM or tetO. Twenty-four of 66 isolates had tetM alone, 21 had tetK alone and 21 had both tetK and tetM (tetKM). All isolates were tetL- and tetO-negative. MICs of tetracycline, doxycycline and minocycline were evaluated for all isolates with or without preincubation in the presence of subinhibitory concentrations of tetracycline or minocycline. All isolates with one or more tetracycline resistance determinants were resistant to tetracycline 8 mg/L without induction of resistance. Some MRSA isolates of each of these three genotypes showed an unexpected lack of resistance to tetracyclines when the disc diffusion or agar dilution method was applied to uninduced cells. Resistance to tetracycline and doxycycline was greater (two- to four-fold) in tetK cells preincubated with tetracycline (tetK MRSA isolates were susceptible to minocycline Phe; Glu87-->Lys), and with a mutation in the parC gene (Ser79-->Ile). However, different mutations in the gyrA gene (Ser83-->Tyr) and parC gene (Ser79-->Phe) were found in a S. sanguis isolate exposed to ofloxacin. A fluoroquinolone-resistant isolate, QR 95101, from a dental infection, had a single mutation in the gyrA gene (Ser83- >Phe) and in the parC gene (Ser79-->Phe). Two fluoroquinolone-resistant mutants, QS-701OFm and QS-701DUm, were obtained from S. anginosus QS-701, by exposure to ofloxacin and DU-6859a, respectively. These mutants showed a common substitution at codon 83 (Ser-->Phe) in the gyrA gene but had different substitutions at codon 87 (QS-701OFm, Glu-->Gln; QS-701DUm, Glu-->Lys). They also had different substitutions at codons 79 and 135 in the parC gene (QS-701OFm, Ser79-->Leu but no change at Glu135; QS-701DUm, Ser79-->Ile and Glu135-->Gln). The resistance levels of the DU-6859a-selected resistant S. sanguis mutant QS-951DUm to DU 6859a, ofloxacin, ciprofloxacin and norfloxacin were higher than those of the ofloxacin-selected resistant mutant QS-951OFm. However, ampicillin susceptibilities of these mutants were not different from the parental strains. In S. anginosus, the DU-6859a-selected fluoroquinolone-resistant mutant QS-701DUm was resistant to all the fluoroquinolones tested, while the ofloxacin-selected mutant QS-701OFm was resistant to three fluoroquinolones, but not DU-6859a. The results indicate that different fluoroquinolones select distinct mutations in the QRDR of the gyrA and parC genes in oral streptococci. The gyrA or parC mutation in oral streptococci may determine the levels of fluoroquinolone resistance. PMID- 10837429 TI - A multiplex polymerase chain reaction to differentiate beta-lactamase plasmids of neisseria gonorrhoeae. AB - In penicillinase-producing Neisseria gonorrhoeae (PPNG), resistance to penicillin may be mediated by one of several related plasmids of different sizes. These include the Asian, African and Rio/Toronto plasmids. Identification of these plasmids provides useful epidemiological information, but has necessitated plasmid purification and gel analysis. We have developed a rapid, simple multiplex polymerase chain reaction (PCR) which discriminates between the beta lactamase resistance plasmids that are frequently found in strains of N. gonorrhoeae. Amplicons of 1191, 958 and 650 bp were produced from strains containing the African, Asian and Rio/Toronto plasmids, respectively, whilst no products resulted from non-PPNG strains harbouring the cryptic, conjugative or tetracycline resistance plasmids. PCR analysis of 123 strains of PPNG identified 60 strains with African, 16 strains with Asian and 47 strains with Rio/Toronto plasmids and showed complete agreement with the standard plasmid analysis. PMID- 10837430 TI - Analysis of the effects of -42 and -32 ampC promoter mutations in clinical isolates of Escherichia coli hyperproducing ampC. AB - Escherichia coli usually produces only very small amounts of a constitutive AmpC beta-lactamase, but clinical strains overproducing this enzyme have been isolated. Three different ampC promoters of E. coli clinical strains were cloned upstream of the chloramphenicol acetyltransferase (CAT) gene in the pKK232-8 reporter plasmid and their relative strengths were compared by two different methods. The strength of the promoters from AmpC hyperproducers was 70- to 120 fold higher than those from a low-level AmpC producer. One of the strong promoters, which differs from strain K12 at bases -88, -82, -42, -18, -1 and +58, was mutated to abolish the -42 mutation. This change resulted in a 43-fold decrease in CAT concentration. In another promoter, with eight different mutations at positions -88, -82, -32, -18, -1, +5, +24 and +58, the -32T-->A transversion, which created perfect homology with the -35 consensus sequence, was reverted; this led to a 13-fold decrease in CAT concentration. The -42 and -32 mutations play an important role in E. coli resistance to beta-lactams by increasing ampC transcription. PMID- 10837431 TI - An evaluation of the potential of the multiple antibiotic resistance operon (mar) and the multidrug efflux pump acrAB to moderate resistance towards ciprofloxacin in Escherichia coli biofilms. AB - The chromosomal multiple antibiotic resistance operon, mar, is widely represented amongst Gram-negative bacteria and has been implicated in resistance towards oxidative stress agents, organic solvents and a large number of structurally unrelated antimicrobial agents. The major mechanism associated with such increased resistance is an upregulation of the efflux pump acrAB. Growth as a biofilm is often associated with similar generalized reductions in susceptibility to inimical agents. Escherichia coli K12 (AG100), an isogenic mutant of AG100 constitutive for mar expression (AG102) and an isolate deleted of the mar locus (MCH164) were grown as biofilms in cellulose-fibre depth filters and perfused with a simple salts, minimal medium (CDM) over 120 h. Biofilms were exposed to various concentrations of ciprofloxacin (0.004, 0.015 and 0.1 mg/L) for 42 h. The numbers of viable cells within the perfusate and within the biofilm were estimated throughout. Whereas no differences were seen between the wild-type and mar-deleted isolates, that constitutive for mar displayed reduced susceptibility to ciprofloxacin at concentrations of 0.004 mg/L (MIC for AG100 was 0.0052 mg/L). Similar antibiotic perfusion experiments were conducted using isolates in which the efflux pump acrAB was either deleted (AG100-A) or constitutively expressed (AG100-B). Exposure of AG100-A biofilms to ciprofloxacin at 0.004 and 0.1 mg/L showed similar susceptibilities to those seen in the wild-type (AG100) and mar deleted (MCH164) isolates and suggested that acrAB was not induced within the attached population. On the other hand, constitutive expression of acrAB (AG100 B) protected biofilms against the lower concentration of ciprofloxacin used (0.004 mg/L). This protection was again lost at concentrations of 0.1 mg/L. Overall, these results show that ciprofloxacin resistance in biofilms is not mediated by the upregulation of the mar or acrAB operons. PMID- 10837432 TI - Activity of linezolid against Gram-positive cocci possessing genes conferring resistance to protein synthesis inhibitors. AB - Linezolid belongs to a new class of antimicrobials, the oxazolidinones, that act by inhibiting protein synthesis. To detect cross-resistance with other inhibitors of protein synthesis (chloramphenicol, macrolides, lincosamides, streptogramins, aminoglycosides and tetracyclines), the in vitro activity of linezolid was determined against isolates harbouring known genes conferring resistance to these antimicrobials. Neither the presence of modifying enzymes (LinA, LinA', LinB, Vgb, Vat, SatA, ANT(4') (4")-I, AAC(6')-APH(2"), APHA-3 and Cat), nor the presence of an efflux mechanism (MsrA, MefE, MefA, MreA, Vga, TetK and TeL), nor the modification or protection of antimicrobial target (because of ribosomal methylases or TetM and TetO) affected linezolid activity as demonstrated by similar in vitro activity against resistant isolates and sensitive control isolates. PMID- 10837433 TI - Trans-epithelial intestinal elimination of moxifloxacin in rabbits. AB - The intestinal trans-epithelial elimination of moxifloxacin was measured in the jejunum, ileum, caecum and in the bile in a rabbit model. Over a 120 min period following a single parenteral administration of moxifloxacin 15 mg/kg, peak serum concentration was 3.1 (+/- 1.1) mg/L. The elimination constants were: 0.019 (+/- 0. 017) microg/cm(2)/min, 0.011 (+/- 0.014) microg/cm(2)/min and 0.002 (+/- 0.002) microg/cm(2)/min in the jejunum, ileum and caecum, respectively. Per loop, over 120 min, the respective eliminated quantities were: 9.59 (+/- 9.37) microg, 8.26 (+/- 6.74) microg and 1.92 (+/- 1.86) microg. Biliary moxifloxacin concentrations varied between 1.30 and 5.16 mg/L and exceeded serum levels from 45 min onwards. Intestinal concentrations paralleled serum moxifloxacin levels. Altogether, approximately 4.5% of the moxifloxacin dose was eliminated trans epithelially in the digestive tract over the 120 min experimental period. PMID- 10837434 TI - The 7alpha-methoxy substituent in cephem or oxacephem antibiotics enhances in vivo anti-Helicobacter felis activity in mice after oral administration. AB - The in vivo antibacterial activities of 7alpha-methoxy-cephems or 7alpha-methoxy 1-oxacephems and their demethoxy congeners after oral administration were compared in a mouse model of gastric infection with Helicobacter felis. The MICs of all four pairs of compounds with and without the 7alpha-methoxy substituent were within one two-fold dilution for H. felis and Helicobacter pylori, but the 7alpha-methoxy compounds were at least four-fold more active at bacterial eradication than their demethoxy congeners. Since these compounds are not absorbed after oral administration, they are likely to have gained direct access to the bacteria from the gastric cavity. Thus the 7alpha-methoxy substituent may enhance in vivo activity by promoting the gastric mucus permeability of the compounds or by allowing the compounds to remain longer in the gastric cavity. PMID- 10837435 TI - Comparison of efficacy of ciprofloxacin and doxycycline against experimental melioidosis and glanders. AB - Melioidosis and glanders are caused by the closely related species Burkholderia pseudomallei and Burkholderia mallei, respectively. Whereas melioidosis is a significant cause of morbidity in south-east Asia, glanders is extremely rare. The efficacies of ciprofloxacin and doxycycline were assessed against a strain of B. pseudomallei and a strain of B. mallei which were susceptible to both antimicrobials in vitro. Porton outbred mice and Syrian hamsters were given 40 mg/kg of either doxycycline or ciprofloxacin twice daily by sc injection according to one of three regimens: dosing starting 48 h before challenge and continuing for 5 days postchallenge; 5 days' therapy starting immediately after challenge; 5 days' therapy starting 24 h after challenge. Mice were challenged ip with B. pseudomallei 4845 and hamsters were challenged ip with B. mallei 23344. Antimicrobial efficacy was determined by the shift in the median lethal dose (MLD). Ciprofloxacin prophylaxis and immediate therapy both raised the MLD of B. pseudomallei to 4 x 10(6) cfu from 19 cfu in untreated animals, but therapeutic ciprofloxacin only raised the MLD to 180 cfu. The results for doxycycline were similar. Ciprofloxacin prophylaxis raised the MLD of B. mallei 23344 to 4.6 x 10(5) cfu compared with 4 cfu in untreated controls. Immediate therapy raised the MLD to 7.0 x 10(4) cfu and therapy raised the MLD to 1.6 x 10(3) cfu. All regimens of doxycycline protected hamsters against challenges of up to 2 x 10(7) cfu. Despite using a susceptible strain of B. pseudomallei, neither antimicrobial was effective when used therapeutically. The timely administration of either antimicrobial, however, was effective in preventing symptomatic infection. Doxycycline was the superior of the two antimicrobials against experimental glanders although relapse did occur in treated animals approximately 4-5 weeks after challenge. PMID- 10837436 TI - A combination of benznidazole and ketoconazole enhances efficacy of chemotherapy of experimental Chagas' disease. AB - Despite the large number of patients infected with Trypanosoma cruzi, there are no commercial drugs available with high efficacy for use in the clinical treatment of Chagas' disease (American trypanosomiasis). As the prospects of the introduction of new compounds by the pharmaceutical industry are poor, alternative strategies are being designed to identify candidates among drugs already available on the market that could be used in combination to provide a synergic effect and improve the efficacy of chemotherapy. In this paper we investigated a possible synergic effect of drugs in mice inoculated with isolates of T. cruzi susceptible (CL), moderately resistant (Y) and naturally resistant (Colombiana) to benznidazole and nifurtimox. Our data demonstrated that the combination of benznidazole with ketoconazole induced a synergic effect in mice infected with the CL and Y isolate of T. cruzi. No differences were observed, however, in animals infected with the Colombiana isolate, suggesting that the synergic effect of benznidazole and ketoconazole is influenced by the isolate of parasite and that this could be important in further studies searching for useful combinations of drugs. Moreover, we observed that early treatment with ketoconazole could increase the cure rate in animals infected with the Y isolate. No positive effect, in relation to cure rate, was observed with the combination of benznidazole and ofloxacin. Our results re-emphasize the importance of identifying those compounds already on the market with synergic effects able to enhance the cure of T. cruzi infection. PMID- 10837437 TI - Further evidence from a murine infection model that famciclovir interferes with the establishment of HSV-1 latent infections. AB - Mice were infected with herpes simplex virus type 1 (HSV-1) via the ear pinna. Famciclovir therapy was commenced on days 2-7 post infection (p.i.). The ipsilateral and contralateral trigeminal (TG) and third cervical ganglia (CIII) from individual mice were tested for latency 1 and 6 months after infection by explant culture or in situ hybridization for latency-associated transcripts (LAT). There were significantly fewer LAT-positive neurons in ipsilateral and contralateral TG (but not CIII) when therapy was delayed by up to 6 days. There was a low correlation between the number of LAT-positive neurons and reactivation by explant culture. Latency data for individual ganglia, compared with those from previous studies, allow us to rationalize differences between the effects of nucleosides on the establishment of latency in different anatomical sites and when tissues are evaluated using different techniques. The implications of the findings for the use of famciclovir to counter HSV latency in humans are addressed. PMID- 10837438 TI - Teicoplanin therapy for Staphylococcus aureus septicaemia: relationship between pre-dose serum concentrations and outcome. AB - Logistic regression analysis was performed on data drawn from a clinical trials database for Staphylococcus aureus septicaemia treated with teicoplanin. Variables analysed were age, body weight, mean pre-dose and post-dose serum teicoplanin concentrations, mean dose (mg or mg/kg body weight) and combination versus monotherapy. Only two variables correlated with clinical outcome at a significance level better than 0.05: age (P = 0.012) and mean pre-dose serum concentration (P = 0.010). The probability of successful treatment declined with age and increased with mean pre-dose serum concentration. PMID- 10837439 TI - A prospective, randomized, double-blinded, placebo-controlled trial of empirical teicoplanin in febrile neutropenia with persistent fever after imipenem monotherapy. AB - Glycopeptide antibiotics are used extensively in the empirical treatment of febrile patients with neutropenia. To come to a more rational and restricted application of these expensive drugs and to reduce the risk of emergence of resistance, we carried out a prospective, double-blinded, placebo-controlled single-centre study to investigate whether the addition of teicoplanin improved the outcome of neutropenic patients who remained febrile after 72-96 h of imipenem monotherapy. Patients with known infections caused by imipenem-resistant microorganisms were excluded. From the 114 evaluable episodes (out of a total of 125) in 105 patients who met the eligibility criteria, 56 episodes were randomized to receive teicoplanin and 58 to placebo. At 72 h after the start of the assigned intervention, 52 (45.6%) of the patients were afebrile; at the end of the aplastic phase, 10 (8.8%) had succumbed. There was no difference between the two study arms. When febrile episodes were subdivided between microbiologically documented infections, clinically documented infections and fevers of unknown origin, again no significant differences were observed. With the exception of methicillin-resistant bacteria, Gram-positive infections seemed to respond well to imipenem monotherapy. It is concluded that the addition of teicoplanin on empirical grounds, i.e. for persistent fever only, is not necessary and that the use of glycopeptides should be restricted to well-defined clinical situations where methicillin-resistant bacteria are involved. Furthermore, it seems that many neutropenic patients respond slowly over more than 72-96 h even when they are treated with antibacterial drugs such as imipenem that are effective against the causative microorganism. PMID- 10837440 TI - Comparison of a 5 day regimen of cefdinir with a 10 day regimen of cefprozil for treatment of acute exacerbations of chronic bronchitis. AB - Patients with acute exacerbations of chronic bronchitis were treated with cefdinir 300 mg bd for 5 days or cefprozil 500 mg bd for 10 days in a prospective, randomized, double-blind, multicentre study. Of the 548 patients enrolled, 281 (51%) were evaluable. The clinical cure rates at the test-of-cure visit were 80% (114/142) and 72% (100/139) for the evaluable patients treated with cefdinir and cefprozil, respectively. Respiratory tract pathogens were isolated from 409 (75%) of 548 admission sputum specimens, with the predominant pathogens being Haemophilus parainfluenzae, Haemophilus influenzae, Staphylococcus aureus and Moraxella catarrhalis. The microbiological eradication rates at the test-of-cure visit were 81% (157 of 193 pathogens) and 84% (166 of 198 pathogens) for the evaluable patients treated with cefdinir and cefprozil, respectively. Adverse event rates while on treatment were equivalent between the two treatment groups. The incidence of diarrhoea during therapy was higher for patients treated with cefdinir (17%) than for patients treated with cefprozil (6%) (P < 0.01), but most cases were mild and did not lead to discontinuation of treatment. These results indicate that a 5 day regimen of cefdinir is as effective and safe in the treatment of patients with acute exacerbations of chronic bronchitis as a 10 day regimen of cefprozil. PMID- 10837441 TI - A multicentre study of the early bactericidal activity of anti-tuberculosis drugs. AB - The early bactericidal activities (EBAs) of 300 mg isoniazid, 18.5 mg isoniazid, 600 mg rifampicin and 800 mg ofloxacin given daily to 262 patients with newly diagnosed pulmonary tuberculosis in Cape Town, Nairobi, Madras and Hong Kong were measured by counting cfu and total acid-fast bacilli in sputum collections taken pre-treatment (S1), at 2 days (S3) and at 5 days (S6). In Cape Town, Nairobi and Madras, the cfu findings suggested that isoniazid produced a massive kill, perhaps of actively growing organisms, during the first 2 days (mean S1-S3 EBAs of 0.636-1.006) but was almost inactive thereafter (mean S3-S6 EBAs of 0.000 0.081), whereas rifampicin maintained moderate activity against slowly growing organisms throughout the 5 days (mean S3-S6 EBAs of 0.242-0.305). This finding suggests that EBAs measured during the 2-5 day interval might be able to assess the sterilizing activity of drugs. Ofloxacin had moderately high mean S1-S3 EBAs of 0.130-0.391. However, in Hong Kong rifampicin appeared to be the most bactericidal drug from the start, possibly because patients had more chronic disease. A method of adjusting the cfu EBAs using total counts was devised which decreased the variability between patients within a treatment group without altering the mean cfu EBA. This resulted in a large gain in precision in Hong Kong, suggesting that their estimates were greatly affected by type II variation, due to dilution of pus by saliva and bronchial secretions, whereas small or no gains were obtained in the other three centres, suggesting that the main cause of variability was type I, due to other factors. PMID- 10837442 TI - Presence of ROB-1 beta-lactamase correlates with cefaclor resistance among recent isolates of Haemophilus influenzae. AB - beta-Lactamase production in Canadian isolates of Haemophilus influenzae has remained relatively constant (25-35%) over the last decade despite increasing cefaclor resistance (MIC >/= 32 mg/L). TEM (294/324, 90.7%) and ROB-1 (30/324, 9.3%) prevalence rates among 324 isolates of H. influenzae obtained from across Canada in 1997-1998 were similar (P > 0.05) to previously published reports. However, 66. 7% (26/39) of cefaclor-resistant isolates were ROB-1-positive (P < 0. 001) and the remaining four ROB-1-positive isolates were cefaclor-intermediate (MIC 16 mg/L). Susceptibilities to loracarbef (P < 0.001) and cefprozil were also reduced in the presence of ROB-1 while the activities of cefuroxime, cefotaxime, cefixime and imipenem were similar in both TEM- and ROB-1-positive solates. PMID- 10837443 TI - beta-lactamase expression in Plesiomonas shigelloides. AB - We have examined the production of beta-lactamases by 11 clinical and nine environmental isolates of Plesiomonas shigelloides from Czechoslovakia, the Czech Republic and Cuba. Ten isolates (50%) expressed detectable amounts of a chromosomally encoded, non-inducible beta-lactamase, though all isolates showed a broadly similar resistance profile: low-level resistance to ampicillin and higher level resistance to carbenicillin. All strains were susceptible to cephalosporins and meropenem. Three clinical isolates expressed a beta-lactamase similar to a class 2c carbenicillinase, with a pI of 5.2 and three expressed an enzyme similar to a class 2d oxacillinase, with a pI of 5.3. The environmental isolates produced a variety of penicillinases, indicating that there is a reservoir of heterogeneous beta-lactamase genes in this species. PMID- 10837444 TI - Detection of extended-spectrum beta-lactamases in members of the family enterobacteriaceae: comparison of the MAST DD test, the double disc and the Etest ESBL. AB - A technically simple method-the MAST double disc (MDD) test-for the detection of extended-spectrum beta-lactamase (ESBL) production by bacteria is described. A wide range of ESBL, non-ESBL and Class 1 beta-lactamase-producing isolates was examined. The MDD test, which uses discs containing ceftazidime and a complementary disc containing ceftazidime and clavulanate and a second pair containing cefotaxime and cefotaxime and clavulanate was compared with the standard double disc diffusion test and an Etest method. Both the Etest and the MDD correctly identified 93% of ESBL producers. The MDD is an inexpensive alternative to current methods for the detection of ESBL production. PMID- 10837445 TI - Retrospective screening for heterogeneous vancomycin resistance in diverse Staphylococcus aureus clones disseminated in French hospitals. AB - Twenty-five different Staphylococcus aureus strains that are widespread in France were screened by various methods for heterogeneous and low-level resistance to vancomycin. Population analysis on brain-heart infusion agar containing 4 mg/L of the drug detected resistant cells at frequencies of 10-7 to 10-6 in five multiply resistant strains. There was no antagonism between vancomycin and beta-lactam antibiotics. One of the five strains, isolated in 1993, was considered a putative progenitor of a French nosocomial S. aureus strain isolated in 1998 and for which the vancomycin MIC was 8 mg/L. PMID- 10837446 TI - Prevalence of macrolide-resistance genes in Staphylococcus aureus and Enterococcus faecium isolates from 24 European university hospitals. AB - The polymerase chain reaction (PCR) was used to study the prevalence of the macrolide resistance genes ermA, ermB, ermC, msrA/msrB, ereA and ereB, in 851 clinical isolates of Staphylococcus aureus and 75 clinical isolates of Enterococcus faecium that were erythromycin resistant. The isolates were from 24 European university hospitals. In S. aureus, the ermA gene was more common in methicillin-resistant S. aureus (MRSA) isolates (88%) than in methicillin susceptible S. aureus (MSSA) isolates (38%), and occurred mainly in strains with constitutive MLS(B) expression. In contrast, ermC was more common in MSSA (47%) than in MRSA (5%), occurring mainly in strains with inducible expression. The ereB gene was only found in MRSA isolates expressing a constitutive MLS(B) phenotype (1%). The ereA gene was not detected. Macrolide resistance by efflux due to the msrA/msrB gene was only detected in MSSA isolates (13%). In contrast to S. aureus, erythromycin resistance in E. faecium was almost exclusively due to the presence of the ermB gene (93%). PMID- 10837447 TI - Antimicrobial resistance in Enterobacteriaceae in Brooklyn, NY: epidemiology and relation to antibiotic usage patterns. AB - In November 1997, all Enterobacteriaceae isolated at 15 hospitals in Brooklyn were collected. Extended-spectrum beta-lactamases (ESBLs) were present in 44% of 409 Klebsiella pneumoniae isolates. Six isolates had reduced susceptibility to carbapenems, including two that were not susceptible to any of the antibiotics tested. Pulsed field gel electrophoresis revealed a commonality of resistant isolates within and between hospitals. The occurrence of ESBLcontaining isolates was associated with cephalosporin usage (P = 0.055). ESBLs were present in 4.7% of Escherichia coli and 9.5% of Proteus mirabilis isolates. It is concluded that ESBL-producing Enterobacteriaceae are endemic in Brooklyn, are spread between hospitals, and may be associated with cephalosporin usage. PMID- 10837448 TI - In vitro activity of meropenem, imipenem, cefepime and ceftazidime against Pseudomonas aeruginosa isolates from cystic fibrosis patients. AB - We studied 67 Pseudomonas aeruginosa isolates from cystic fibrosis patients, and compared their in vitro susceptibility to two carbapenems (meropenem and imipenem) and two cephalosporins (cefepime and ceftazidime). The carbapenems were more effective in vitro than the cephalosporins: 92.5% of isolates were susceptible to the former and 77.6% to the latter. Essentially no difference was found between meropenem and imipenem. More discrepancies were seen between cefepime and ceftazidime: four of 67 isolates (6.0%) were more susceptible to cefepime than to ceftazidime, while eight (11. 9%) were more susceptible to ceftazidime than to cefepime. PMID- 10837449 TI - Observations on the antibiotic treatment of experimentally induced mycoplasmal infections in mice. AB - Antibiotics were given subcutaneously to female mice, colonized by mycoplasmas in the vagina and by others in the oropharynx. Mycoplasma pulmonis was eradicated from the vagina of 36 of 42 immunocompetent TO or BALB/c mice with oxytetracycline, and from 17 of 18 TO mice with lymecycline. Mycoplasma hominis was eradicated from the vagina of all of 18 immunocompetent BALB/c mice with oxytetracycline or tetracycline. Regarding oropharyngeal organisms, M. pulmonis was eradicated from only 20 of 42 TO or BALB/c mice with oxytetracycline and from none of 15 mice with lymecycline. However, Mycoplasma pneumoniae was eliminated from the oropharynx of eight of nine BALB/c mice with oxytetracycline. In contrast to the success in eradicating mycoplasmas from the vagina or oropharynx of immunocompetent BALB/c mice with oxytetracycline, no such effect was seen in nude BALB/c mice, indicating the importance of a competent immune system in conjunction with antibiotic treatment for eradication of these organisms. PMID- 10837450 TI - Equivalence of ceftriaxone and rifampicin in eliminating nasopharyngeal carriage of serogroup B Neisseria meningitidis. AB - The efficacy of ceftriaxone in eliminating nasopharyngeal carriage of Neisseria meningitidis was compared with that of rifampicin during an epidemic of serogroup B meningococcal disease in Auckland, New Zealand. Household contacts of cases had a throat swab taken and were randomized to treatment. Carriers had a repeat swab taken 6 days later to determine efficacy of treatment. Ceftriaxone (98.2%) was equivalent to rifampicin (97.6%) in eliminating serogroup B N. meningitidis. It is cheaper than rifampicin and has the advantage of full compliance and fewer contraindications, but its acceptability by patients may limit its use as a first line prophylactic agent. PMID- 10837451 TI - Trends in hospital antimicrobial prescribing after 9 years of stewardship. AB - Trends in antibiotic prescribing in Grampian have been monitored prospectively for 11 years from 1986 using computerized ward stock lists and laboratory data relating to all in-patient and out-patient treatments in all Grampian hospitals. The main outcome measures were the number of antibiotics available for routine and restricted use, annual expenditure and defined daily doses (DDDs) of high expenditure antimicrobial agents. An antibiotic committee introduced a policy and formulary in the third year of the study which has had only limited success in controlling prescribing. This report updates the audit from 1992/3 to 1996/7. During this period 22 new antibiotics were considered for inclusion in the hospital formulary. Seventeen, including seven antiretroviral agents, were incorporated, all for restricted use only. Despite this, expenditure on antibiotics has more than trebled since 1986/7 and increased 50% since 1992/3, two-thirds of the latter increase being due to the use of new drugs, namely anti HIV drugs, lipid amphotericin derivatives and teicoplanin. Big increases in the use of co-amoxiclav, acyclovir, ciprofloxacin and cefotaxime account for the remainder of the increased expenditure. There was an overall increase of 16.9% in DDDs between 1992/3 and 96/7 to 424.0 DDDs/1000 patient days (393.4 DDDs for antibacterials). These findings highlight the current difficulty in controlling prescribing budgets, the increasing use of antibiotics and the consequent increase of antimicrobial-resistant microorganisms. PMID- 10837452 TI - Towards a common susceptibility testing method? PMID- 10837453 TI - In vitro study of the susceptibility of Escherichia coli to mecillinam. PMID- 10837454 TI - Typing of 154 clinical isolates of Stenotrophomonas maltophilia by pulsed-field gel electrophoresis and determination of the in vitro susceptibilities of these strains to 28 antibiotics. PMID- 10837455 TI - PER-1 beta-lactamase-producing Pseudomonas aeruginosa in an intensive care unit. PMID- 10837456 TI - Quantifying the impact of resistance for prescribers and drug developers-a function of natural resolution rates. PMID- 10837457 TI - Clinafloxacin serum concentrations in a patient on continuous veno-venous haemofiltration. PMID- 10837458 TI - Abnormalities in liver enzymes during simultaneous therapy with itraconazole and amphotericin B in leukaemic patients. PMID- 10837459 TI - Is a 5 day course of azithromycin enough for infections caused by Legionella pneumophila? PMID- 10837460 TI - Two RGD-independent alpha vbeta 3 integrin binding sites on tumstatin regulate distinct anti-tumor properties. AB - Vascular basement membrane is an important regulator of angiogenesis and undergoes many alterations during angiogenesis and these changes are speculated to influence neovascularization. Recently, fragments of collagen molecules have been identified to possess anti-angiogenic activity. Tumstatin (alpha3(IV)NC1 domain) is one such novel molecule with distinct anti-tumor properties and possesses an N-terminal (amino acids 54-132) anti-angiogenic and a C-terminal (amino acids 185-203) anti-tumor cell activity (Maeshima, Y., et al. 2000) J. Biol. Chem. 275, 21340-21348). Previous studies have identified the 185-203 amino acid sequence as a ligand for alpha(v)beta(3) integrin (Shahan, T. A., et al. (1999) Cancer Res. 59, 4584-4590). In the present study, we found distinct additional RGD-independent alpha(v)beta(3) integrin binding site within 54-132 amino acids of tumstatin. This site is not essential for inhibition of tumor cell proliferation but necessary for the anti-angiogenic activity. A fragment of tumstatin containing 54-132 amino acid (tum-2) binds both endothelial cells and melanoma cells but only inhibited proliferation of endothelial cells, with no effect on tumor cell proliferation. A similar experiment with fragment of tumstatin containing the 185-203 amino acid (tum-4) demonstrates that it binds both endothelial cells and melanoma cells but only inhibits the proliferation of melanoma cells. The presence of cyclic RGD peptides did not affect the alpha(v)beta(3) integrin-mediated activity of tumstatin, although significant inhibition of endothelial cell binding to vitronectin was observed. The two distinct RGD-independent binding sites on tumstatin suggest unique alpha(v)beta(3) integrin-mediated mechanisms governing the two distinct anti tumor properties of tumstatin. PMID- 10837461 TI - Mitochondria localization and dimerization are required for CIDE-B to induce apoptosis. AB - Cell death-inducing DFF45-like effector (CIDE)-B is a member of the novel family of apoptosis-inducing factors that share homology with the N-terminal region of DFF, the DNA fragmentation factor. The molecular mechanism of CIDE-B-induced apoptosis is unclear. We have shown here that CIDE-B protein is localized in mitochondria and forms homodimers and heterodimers with other family members. Serial deletion analyses suggest that the mitochondria localization signal and dimerization interface are overlapped and localized to the 30 amino acid residues at the C-terminal region of CIDE-B. Mitochondria localization and dimerization are both required for CIDE-B-induced apoptosis. Our study has thus revealed a mechanism for CIDE-B-induced apoptosis by localization to mitochondria and the formation of a high affinity homo- or heterodimeric complex. PMID- 10837462 TI - The beta 1,3-galactosyltransferase beta 3GalT-V is a stage-specific embryonic antigen-3 (SSEA-3) synthase. AB - We have previously reported the molecular cloning of beta1, 3 galactosyltransferase-V (beta3GalT-V), which catalyzes the transfer of Gal to GlcNAc-based acceptors with a preference for the core3 O-linked glycan GlcNAc(beta1,3)GalNAc structure. Further characterization indicated that the recombinant beta3GalT-V enzyme expressed in Sf9 insect cells also utilized the glycolipid Lc3Cer as an efficient acceptor. Surprisingly, we also found that beta3GalT-V catalyzes the transfer of Gal to the terminal GalNAc unit of the globoside Gb4, thereby synthesizing the glycolipid Gb5, also known as the stage specific embryonic antigen-3 (SSEA-3). The SSEA-3 synthase activity of beta3GalT V was confirmed in vivo by stable expression of the human beta3GalT-V gene in F9 mouse teratocarcinoma cells, as detected with the monoclonal antibody MC-631 by flow cytometry analysis and immunostaining of extracted glycolipids. The biological relation between SSEA-3 formation and beta3GalT-V was further documented by showing that F9 cells treated with the differentiation-inducing agent retinoic acid induced the expression of both the SSEA-3 epitope and the endogenous mouse beta3GalT-V gene. This study represents the first example of a glycosyltransferase, which utilizes two kinds of sugar acceptor substrates without requiring any additional modifier molecule. PMID- 10837463 TI - Transfer of iron-sulfur cluster from NifU to apoferredoxin. AB - Iron-sulfur proteins are present in a wide variety of organisms and are known to play important physiological roles, not only in electron transfer and metabolic reactions, but also in transcriptional regulation. However, little is known about how iron-sulfur clusters themselves are synthesized and assembled within polypeptides. Here we show that a [2Fe-2S] cluster-containing NifU of cyanobacterium Synechocystis PCC6803, SyNifU, possesses the ability to deliver its [2Fe-2S] cluster to an apoferredoxin without the aid of other proteinaceous or nonproteinaceous factor(s). Upon delivery the reconstituted holoferredoxin regained electron transfer ability. The [2Fe-2S] cluster contained within SyNifU was labile upon exposure to the iron-chelating reagent EDTA, suggesting that the iron-sulfur cluster is abnormally exposed to solvent. We propose that NifU serves as a scaffold for iron-sulfur cluster assembly and functions as a mediator for iron-sulfur cluster delivery. PMID- 10837464 TI - Rab15 differentially regulates early endocytic trafficking. AB - Rab GTPases play an important regulatory role in early endocytosis. We recently demonstrated that epitope-tagged Rab15 (HArab15) co-localizes with Rab4, -5, and 11 on early endosomal membranes in CHO cells (Zuk, P. A., and Elferink, L. A. (1999) J. Biol. Chem. 274, 22303-22312). To characterize the role of Rab15 in endocytosis, we prepared functional mutants of HArab15 and examined their effects on early endocytic trafficking. Wild-type HArab15 and its constitutively active, GTP-bound mutant (Q67L) reduce fluid phase and receptor-mediated endocytosis without affecting the rate of recycling from early endosomal compartments. Inhibition of early endocytosis appears to be due to a reduction in the rate of homotypic early endosome fusion. Conversely, mutations that constitutively inactivate HArab15 stimulate early endocytosis and the homotypic fusion of early endosomes in vitro. Unlike active forms of HArab15, constitutively inactive HArab15 mutants also affect recycling from early endosomal compartments. Moreover, the two constitutively inactive mutants, GDP-bound HArab15-T22N and the non-nucleotide binding mutant HArab15-N121I, differentially regulate the transit of fluid phase and receptor-mediated endocytic tracers through early/sorting endosomes. Together, these data suggest that HArab15 may counteract the reported stimulatory effect of Rab5 on early endocytosis. Consistent with this, overexpression of constitutively active HArab15-Q67L attenuates Rab5-stimulated endocytosis, whereas Rab5-stimulated endocytosis is augmented in cells overexpressing a constitutively inactive HArab15 mutant defective in guanine nucleotide binding (N121I). Our data indicate that HArab15 differentially regulates distinct steps in membrane trafficking through early/sorting and pericentriolar recycling endosomes. PMID- 10837465 TI - The NF-kappa B signaling pathway is not required for Fas ligand gene induction but mediates protection from activation-induced cell death. AB - Stimulation of T cells by antigens or mitogens triggers multiple signaling pathways leading to activation of genes encoding interleukin-2 and other growth regulatory cytokines. The same stimuli also activate the gene encoding an apoptosis-inducing molecule, Fas ligand (FasL), which contributes to activation induced cell death. It has been proposed that the signaling pathways involved in cytokine gene induction also contribute to activation-induced FasL expression; however, genetic evidence for this proposal is lacking. In the present study, the role of the NF-kappaB signaling pathway in FasL gene expression was examined using a mutant T cell line deficient in an essential NF-kappaB signaling component, IkappaB kinase gamma. These mutant cells have a blockade in signal induced activation of NF-kappaB but remained normal in the activation of NF-AT and AP-1 transcription factors. Interestingly, the NF-kappaB signaling defect has no effect on mitogen-stimulated FasL gene expression, although it completely blocks the interleukin-2 gene induction. We further demonstrate that NF-kappaB activation is required for protecting T cells from apoptosis induction by mitogens and an agonistic anti-Fas antibody. These genetic results suggest that the NF-kappaB signaling pathway is not required for activation-induced FasL expression but rather mediates cell growth and protection from activation-induced cell death. PMID- 10837466 TI - Theromin, a novel leech thrombin inhibitor. AB - We purified the most potent thrombin inhibitor described to date from the rhynchobdellid leech Theromyzon tessulatum. Designated theromin, it was purified to apparent homogeneity by gel permeation and anion exchange chromatography followed by two reverse-phase steps of high performance liquid chromatography. The primary sequence of theromin (a homodimer of 67 amino acid residues including 16 cysteine residues) was determined by a combination of reduction and s-beta pyridylethylation, Edman degradation, trypsin enzymatic digestion, and matrix assisted laser desorption mass spectrometry measurement. Theromin exhibits no sequence homology with any other thrombin inhibitors. Furthermore, theromin significantly diminishes, in a dose-dependent manner, the level of human granulocyte and monocyte activation induced by lipopolysaccharides. In summary, this potent thrombin inhibitor promises to have high biomedical significance. PMID- 10837467 TI - Refolding of target proteins from a "rigid" mutant chaperonin demonstrates a minimal mechanism of chaperonin binding and release. AB - One of the most interesting facets of GroEL-facilitated protein folding lies in the fact that the requirement for a successful folding reaction of a given protein target depends upon the refolding conditions used. In this report, we utilize a mutant of GroEL (GroEL T89W) whose domain movements have been drastically restricted, producing a chaperonin that is incapable of utilizing the conventional cyclic mechanism of chaperonin action. This mutant was, however, still capable of improving the refolding yield of lactate dehydrogenase in the absence of both GroES and ATP hydrolysis. A very rapid interconversion of conformations was detected in the mutant immediately after ATP binding, and this interconversion was inferred to form part of the target release mechanism in this mutant. The possibility exists that some target proteins, although dependent on GroEL for improved refolding yields, are capable of refolding successfully by utilizing only portions of the entire mechanism provided by the chaperonins. PMID- 10837468 TI - Aut7p, a soluble autophagic factor, participates in multiple membrane trafficking processes. AB - Aut7p, a protein recently implicated in autophagic events in the yeast Saccharomyces cerevisiae, exhibits significant homology to a mammalian protein, p16, herein termed GATE-16 (Golgi-associated ATPase Enhancer of 16 kDa), a novel intra-Golgi transport factor. Here we provide evidence for the involvement of Aut7p in different membrane trafficking processes. Aut7p largely substitutes for the activity of GATE-16 in mammalian intra-Golgi transport in vitro. In vivo, AUT7 interacts genetically with endoplasmic reticulum to Golgi SNAREs, specifically with BET1 and SEC22. Aut7p interacts physically with the following two v-SNAREs: Bet1p, which is involved in endoplasmic reticulum to Golgi vesicular transport, and Nyv1p, implicated in vacuolar inheritance. We suggest that, in addition to its role in autophagocytosis, Aut7p has pleiotropic effects and participates in at least two membrane traffic events. PMID- 10837469 TI - Crystal structure of human parathyroid hormone 1-34 at 0.9-A resolution. AB - The N-terminal fragment 1-34 of parathyroid hormone (PTH), administered intermittently, results in increased bone formation in patients with osteoporosis. PTH and a related molecule, parathyroid hormone-related peptide (PTHrP), act on cells via a common PTH/PTHrP receptor. To define more precisely the ligand-receptor interactions, we have crystallized human PTH (hPTH)-(1-34) and determined the structure to 0.9-A resolution. hPTH-(1-34) crystallizes as a slightly bent, long helical dimer. Analysis reveals that the extended helical conformation of hPTH-(1-34) is the likely bioactive conformation. We have developed molecular models for the interaction of hPTH-(1-34) and hPTHrP-(1-34) with the PTH/PTHrP receptor. A receptor binding pocket for the N terminus of hPTH (1-34) and a hydrophobic interface with the receptor for the C terminus of hPTH (1-34) are proposed. PMID- 10837470 TI - p38 mitogen-activated protein kinase mediates bid cleavage, mitochondrial dysfunction, and caspase-3 activation during apoptosis induced by singlet oxygen but not by hydrogen peroxide. AB - p38 mitogen-activated protein kinase is activated and involved in cleavage of caspase-3 during apoptosis induced by a number of stimuli. However, the signaling events triggered by p38 that result in caspase-3 activation are still unknown. In human leukemia cells, two reactive oxygen species, singlet oxygen and hydrogen peroxide (H(2)O(2)), selectively stimulated the phosphorylation of p38. Preincubation of cells with SB203580, a specific inhibitor of p38, dose dependently inhibited DNA fragmentation induced by singlet oxygen but not by H(2)O(2). Protection from apoptosis by SB203580 correlated with inhibition of caspase-3, and several events that are associated with caspase-3 activation, including Bid cleavage, decrease in mitochondrial transmembrane potential and release of cytochrome c from mitochondria, whereas caspase-8 cleavage was not affected by this inhibitor. In contrast, blockade of caspase-8 with Ile-Glu-Thr Asp-fluoromethyl ketone is sufficient to prevent formation of DNA fragments and to inhibit all the above signaling events, with exception of p38 phosphorylation, in both singlet oxygen- and H(2)O(2)-treated cells. These data suggest that caspase-3 activation is regulated through redundant signaling pathways that involve p38 and caspase-8 acting upstream of Bid during singlet oxygen-induced apoptosis, whereas the activation of caspase-3 by H(2)O(2) is only governed by a caspase-8-mediated apoptotic pathway. PMID- 10837471 TI - The role of alpha and beta chains in ligand recognition by beta 7 integrins. AB - Integrins alpha(E)beta(7) and alpha(4)beta(7) are involved in localization of leukocytes at mucosal sites. Although both alpha(E)beta(7) and alpha(4)beta(7) utilize the beta(7) chain, they have distinct binding specificities for E cadherin and mucosal addressin cell adhesion molecule-1 (MAdCAM-1), respectively. We found that mutation of the metal ion-dependent adhesion site (MIDAS) in the alpha(E) A-domain (D190A) abolished E-cadherin binding, as did mutation F298A on the A-domain surface near the MIDAS cleft. A docking model of the A-domain with E cadherin domain 1 indicates that coordination of the alpha(E) MIDAS metal ion by E-cadherin Glu(31) and a novel projection of Phe(298) into a hydrophobic pocket on E-cadherin provide the basis for the interaction. The location of the binding site on the alpha(E) A-domain resembles that on other integrins, but its structure appears distinctive and particularly adapted to recognize the tip of E cadherin, a unique integrin ligand. Additionally, mutation of the beta(7) MIDAS motif (D140A) abolished alpha(E)beta(7) binding to E-cadherin and alpha(4)beta(7) mediated adhesion to MAdCAM-1, and alpha(4) chain mutations that abrogated binding of alpha(4)beta(1) to vascular cell adhesion molecule-1 and fibronectin similarly reduced alpha(4)beta(7) interaction with MAdCAM-1. Thus, although specificity can be determined by the integrin alpha or beta chain, common structural features of both subunits are required for recognition of dissimilar ligands. PMID- 10837472 TI - Mechanism of inhibition of the class A beta -lactamases PC1 and TEM-1 by tazobactam. Observation of reaction products by electrospray ionization mass spectrometry. AB - The reactions of class A beta-lactamases PC1 and TEM-1 with tazobactam (TZB), a potent penicillanic sulfone inhibitor for class A beta-lactamases, were studied using electrospray ionization mass spectrometry (ESI/MS). Following inactivation of the beta-lactamases by TZB, new abundant high mass components were observed including three with molecular masses of 52, 70, and 88 Da greater than PC1 and TEM-1, respectively, and a component with a molecular mass of 300 Da greater than PC1. In addition, three TZB reaction products with molecular masses of 248, 264, and 280 Da were observed. High performance liquid chromatography (HPLC)/ESI/MS analysis of the TZB-PC1 adduct digested with Glu-C revealed three new components with masses 52, 70, and 88 Da greater than that of the peptide composed of amino acid residues 58-82 and one new component with a mass 70 Da greater than that of the peptide composed of amino acid residues 125-141. HPLC/ESI/MS/MS analysis of the two digested peptides whose masses increased by 70 Da indicated that Ser-70 and Ser-130 were the most likely TZB-modified amino acid residues. Based on these data, a mechanism for the inactivation of the class A beta-lactamases by TZB is proposed. In this scheme, initial acylation of Ser-70 by TZB and opening of the lactam ring are followed by one of several different events: (1) the rapid decomposition of TZB with loss of the enamine moiety to form the propiolylated enzyme, (2) an intramolecular nucleophilic displacement of the imine or enamine moiety by Ser-130 to form a cross-linked vinyl ether, and (3) hydrolysis of the imine or enamines to form a Ser-70-linked aldehyde. PMID- 10837473 TI - Growth factors inactivate the cell death promoter BAD by phosphorylation of its BH3 domain on Ser155. AB - The Bcl-2 family protein BAD promotes apoptosis by binding through its BH3 domain to Bcl-x(L) and related cell death suppressors. When BAD is phosphorylated on either Ser(112) or Ser(136), it forms a complex with 14-3-3 in the cytosol and no longer interacts with Bcl-x(L) at the mitochondria. Here we show that phosphorylation of a distinct site Ser(155), which is at the center of the BAD BH3 domain, directly suppressed the pro-apoptotic function of BAD by eliminating its affinity for Bcl-x(L). Protein kinase A functioned as a BAD Ser(155) kinase both in vitro and in cells. BAD Ser(155) was found to be a major site of phosphorylation induced following stimulation by growth factors and prevented by protein kinase A inhibitors but not by inhibitors of the phosphatidylinositol 3 kinase/Akt pathway. Growth factors inhibited BAD-induced apoptosis in both a Ser(112)/Ser(136)- and a Ser(155)-dependent fashion. Thus, growth factors engage an anti-apoptotic signaling pathway that inactivates BAD by direct modification of its BH3 cell death effector domain. PMID- 10837475 TI - Characterization of YpmQ, an accessory protein required for the expression of cytochrome c oxidase in Bacillus subtilis. AB - A search of the Bacillus subtilis genome identifies a potential homolog, ypmQ, of the inner mitochondrial membrane protein Sco1 from yeast. Sco1 has been found to aid the delivery of copper to cytochrome c oxidase. B. subtilis expresses two members of the cytochrome oxidase family, a cytochrome c oxidase that has two copper centers, Cu(A) and Cu(B), and a menaquinol oxidase that has only Cu(B). Deletion of ypmQ in B. subtilis depresses expression of cytochrome c oxidase but not menaquinol oxidase. Levels of cytochrome c oxidase recover when copper is added to the growth medium of the DeltaypmQ strain or when ypmQ is expressed from a plasmid. Neither treatment affects the amount or activity of menaquinol oxidase. YpmQ in which two conserved cysteines are replaced by serines and a conserved histidine is replaced by alanine do not complement the deletion of ypmQ even though these mutant forms are found in the membrane extract at a level similar to the wild type protein. We propose that the two cysteines and the histidine are critical for the function of YpmQ and suggest they are involved in copper exchange between YpmQ and the Cu(A) site of cytochrome c oxidase. PMID- 10837476 TI - An analysis of the interaction between mouse apolipoprotein B100 and apolipoprotein(a). AB - The assembly of lipoprotein(a) (Lp(a)) involves an initial noncovalent interaction between apolipoprotein (apo) B100 and apo(a), followed by the formation of a disulfide bond between apoB100 cysteine 4326 and apo(a) cysteine 4057. The structural features of apoB100 that are required for its noncovalent interaction with apo(a) have not been fully defined. To analyze that initial interaction, we tested whether apo(a) could bind noncovalently to two apoB proteins that lack cysteine 4326: mouse apoB100 and human apoB100-C4326G. Our experiments demonstrated that both mouse apoB and the human apoB100-C4326G bind noncovalently to apo(a). We next sought to gain insights into the apoB amino acid sequences required for the interaction between apoB100 and apo(a). Previous studies of truncated human apoB proteins indicated that the carboxyl terminus of human apoB100 (amino acids 4330-4397) is important for Lp(a) assembly. To determine whether the carboxyl terminus of mouse apoB100 can interact with apo(a), transgenic mice were produced with a mutant human apoB gene construct in which human apoB100 amino acids 4279-4536 were replaced with the corresponding mouse apoB100 sequences and tyrosine 4326 was changed to a cysteine. The mutant apoB100 bound to apo(a) and formed bona fide disulfide-linked Lp(a), but Lp(a) assembly was less efficient than with wild-type human apoB100. The fact that Lp(a) assembly was less efficient with the mouse apoB sequences provides additional support for the notion that sequences in the carboxyl terminus of apoB100 are important for Lp(a) assembly. PMID- 10837477 TI - Apg13p and Vac8p are part of a complex of phosphoproteins that are required for cytoplasm to vacuole targeting. AB - We have been studying protein components that function in the cytoplasm to vacuole targeting (Cvt) pathway and the overlapping process of macroautophagy. The Vac8 and Apg13 proteins are required for the import of aminopeptidase I (API) through the Cvt pathway. We have identified a protein-protein interaction between Vac8p and Apg13p by both two-hybrid and co-immunoprecipitation analysis. Subcellular fractionation of API indicates that Vac8p and Apg13p are involved in the vesicle formation step of the Cvt pathway. Kinetic analysis of the Cvt pathway and autophagy indicates that, although Vac8p is essential for Cvt transport, it is less important for autophagy. In vivo phosphorylation experiments demonstrate that both Vac8p and Apg13p are phosphorylated proteins, and Apg13p phosphorylation is regulated by changing nutrient conditions. Although Apg13p interacts with the serine/threonine kinase Apg1p, this protein is not required for phosphorylation of either Vac8p or Apg13p. Subcellular fractionation experiments indicate that Apg13p and a fraction of Apg1p are membrane-associated. Vac8p and Apg13p may be part of a larger protein complex that includes Apg1p and additional interacting proteins. Together, these components may form a protein complex that regulates the conversion between Cvt transport and autophagy in response to changing nutrient conditions. PMID- 10837478 TI - Oxidoreductases in lipoxin A4 metabolic inactivation: a novel role for 15 onoprostaglandin 13-reductase/leukotriene B4 12-hydroxydehydrogenase in inflammation. AB - The lipoxins (LX) are autacoids that act within a local inflammatory milieu to dampen neutrophil recruitment and promote resolution. 15-Hydroxyprostaglandin dehydrogenase (15-PGDH) and 15-oxoprostaglandin 13-reductase, also termed leukotriene B(4) 12-hydroxydehydrogenase (PGR/LTB(4)DH), are two enzymatic activities appreciated for their roles in the metabolism of prostaglandins and LTB(4). Here, we determined whether these oxidoreductases also catalyze the conversion of lipoxin A(4) (LXA(4)) and assessed the activities of these LXA(4) metabolites. 15-Oxo-LXA(4) was generated by incubating LXA(4) with 15-PGDH and NAD(+) for studies of its further conversion. PGR/LTB(4)DH catalyzed the NADH dependent reduction of 15-oxo-LXA(4) to yield 13,14-dihydro-15-oxo-LXA(4). With NADH as a cofactor, 15-PGDH acted as a 15-carbonyl reductase and catalyzed the conversion of 13,14-dihydro-15-oxo-LXA(4) to 13, 14-dihydro-LXA(4). Human polymorphonuclear leukocytes (PMN) exposed to native LXA(4), 15-oxo-LXA(4), or 13,14-dihydro-LXA(4) did not produce superoxide anions. At concentrations where LXA(4) and a metabolically stable LXA(4) analog potently inhibited leukotriene B(4)-induced superoxide anion generation, the further metabolites were devoid of activity. Neither 15-oxo-LXA(4) nor 13, 14-dihydro-LXA(4) effectively competed with (3)H-labeled LXA(4) for specific binding to recombinant LXA(4) receptor (ALXR). In addition, introducing recombinant PGR/LTB(4)DH into a murine exudative model of inflammation increased PMN number by approximately 2-fold, suggesting that this enzyme participates in the regulation of PMN trafficking. These results establish the structures of LXA(4) further metabolites and indicate that conversion of LXA(4) to oxo- and dihydro- products represents a mode of LXA(4) inactivation in inflammation. Moreover, they suggest that these eicosanoid oxidoreductases have multifaceted roles controlling the levels of specific eicosanoids involved in the regulation of inflammation. PMID- 10837479 TI - Each of the three binding sites on complement factor H interacts with a distinct site on C3b. AB - Factor H (fH) restricts activation of the alternative pathway of complement at the level of C3, both in the fluid phase and on self-structures, but allows the activation to proceed on foreign structures. To study the interactions between fH and C3b we used surface plasmon resonance analysis (Biacore(R)) and eight recombinantly expressed fH constructs containing fragments of the 20 short consensus repeat domains (SCRs) of fH. We analyzed the binding of these constructs to C3b and its cleavage products C3c and C3d. Three binding sites for C3b were found on fH. Site 1 was localized to the five amino-terminal SCRs (SCR1 5), and its reciprocal binding site on C3b was found to be lost upon the cleavage of C3b to C3c and C3d. Site 2 on fH was localized by exclusion probably within or near SCRs 12-14 (fragment SCR8-20 bound to C3b, C3c, and C3d; SCR8-11 did not bind to C3b at all; and SCR15-20 bound only to the C3d part of C3b). Site 3 on fH for C3b was localized to the carboxyl-terminal SCRs 19-20, and its reciprocal binding site was mapped to the C3d part of C3b. In conclusion, we confirmed and mapped three binding sites on fH for C3b and demonstrated that the three binding sites on fH interact with distinct sites on C3b. Multiple reciprocal interactions between C3b and fH can provide a basis for the different reactivity of the alternative pathway with different target structures. PMID- 10837480 TI - Molecular anatomy of the peroxin Pex12p: ring finger domain is essential for Pex12p function and interacts with the peroxisome-targeting signal type 1 receptor Pex5p and a ring peroxin, Pex10p. AB - The three peroxin genes, PEX12, PEX2, and PEX10, encode peroxisomal integral membrane proteins with RING finger at the C-terminal part and are responsible for human peroxisome biogenesis disorders. Mutation analysis in PEX12 of Chinese hamster ovary cell mutants revealed a homozygous nonsense mutation at residue Trp263Ter in ZP104 cells and a pair of heterozygous nonsense mutations, Trp170Ter and Trp114Ter, in ZP109. This result and domain mapping of Pex12p showed that RING finger is essential for peroxisome-restoring activity of Pex12p but not necessary for targeting to peroxisomes. The N-terminal region of Pex12p, including amino acid residues at positions 17-76, was required for localization to peroxisomes, while the sequence 17-76 was not sufficient for peroxisomal targeting. Peroxins interacting with RING finger of Pex2p, Pex10p, and Pex12p were investigated by yeast two-hybrid as well as in vitro binding assays. The RING finger of Pex12p bound to Pex10p and the PTS1-receptor Pex5p. Pex10p also interacted with Pex2p and Pex5p in vitro. Moreover, Pex12p was co immunoprecipitated with Pex10p from CHO-K1 cells, where Pex5p was not associated with the Pex12p-Pex10p complex. This observation suggested that Pex5p does not bind to, or only transiently interacts with, Pex10p and Pex12p when Pex10p and Pex12p are in the oligomeric complex in peroxisome membranes. Hence, the RING finger peroxins are most likely to be involved in Pex5p-mediated matrix protein import into peroxisomes. PMID- 10837481 TI - Nonhypoxic pathway mediates the induction of hypoxia-inducible factor 1alpha in vascular smooth muscle cells. AB - Hypoxia-inducible factor-1 (HIF-1) controls the expression of a number of genes such as vascular endothelial growth factor (VEGF) and Erythropoietin in low oxygen conditions (hypoxia). VEGF is strongly induced at both the mRNA and protein expression level by a number of hormones and growth factors in vascular smooth muscle cells (VSMC) independently of the oxygen environment. However, the role of HIF-1alpha in this induction has not been studied. We report here that HIF-1alpha protein levels are strongly increased by fetal calf serum in quiescent VSMC. More interestingly, Angiotensin II (Ang II), thrombin, platelet-derived growth factor, and other hormones can also increase HIF-1alpha in VSMC to levels that are substantially more elevated than the hypoxic treatment. HIF-1alpha induced by Ang II is located in the nucleus, binds to the hypoxic response element, and is transcriptionally active. The induction of HIF-1alpha by hormones is mediated through the production of reactive oxygen species (ROS), since it can be blocked by the ROS inhibitors, diphenyleneiodonium and catalase. Finally, strong induction of VEGF mRNA by Ang II can also be inhibited by these ROS inhibitors. These results implicate HIF-1alpha and HIF-1-dependent transcriptional activity in the induction of VEGF expression after agonist stimulation and define novel hypoxia-independent mechanisms that should play a major role in vascular remodeling. PMID- 10837482 TI - N-Linked oligosaccharides on the meprin A metalloprotease are important for secretion and enzymatic activity, but not for apical targeting. AB - The alpha and beta subunits of meprins, mammalian zinc metalloendopeptidases, are extensively glycosylated; approximately 25% of the total molecular mass of the subunits is carbohydrate. The aim of this study was to investigate the roles of the N-linked oligosaccharides on the secreted form of mouse meprin A. Recombinant meprin alpha and mutants in which one of the 10 potential Asn glycosylation sites was mutated to Gln were all secreted and sorted exclusively into the apical medium of polarized Madin-Darby canine kidney cells, indicating that no specific N-linked oligosaccharide acts as a determinant for apical targeting of meprin alpha. Several of the mutant proteins had decreased enzymatic activity using a bradykinin analog as substrate, and deglycosylation of the wild-type protein resulted in loss of 75-100% activity. Some of the mutants were also more sensitive to heat inactivation. In studies with agents that inhibit glycosylation processes in vivo, tunicamycin markedly decreased secretion of meprin, whereas castanospermine and swainsonine had little effect on secretion, sorting, or enzymatic properties of meprin. When all the potential glycosylation sites on a truncated form of meprin alpha (alpha-(1-445)) were mutated, the protein was not secreted into the medium, but was retained within the cells even after 10 h. These results indicate that there is no one specific glycosylation site or type of oligosaccharide (high mannose- or complex-type) that determines apical sorting, but that core N-linked carbohydrates are required for optimal enzymatic activity and for secretion of meprin alpha. PMID- 10837483 TI - Crocus sativus lectin recognizes Man3GlcNAc in the N-glycan core structure. AB - Crocus sativus lectin (CSL) is one of the truly mannose-specific plant lectins that has a unique binding specificity that sets it apart from others. We studied sugar-binding specificity of CSL in detail by a solution phase method (fluorescence polarization) and three solid phase methods (flow injection, surface plasmon resonance, and microtiter plate), using a number of different glycopeptides and oligosaccharides. CSL binds the branched mannotriose structure in the N-glycan core. Substitution of the terminal Man in the Manalpha(1-3)Man branch with GlcNAc drastically decreases binding affinity much more than masking of the terminal Man in the Manalpha(1-6)Man branch. Most interestingly, the beta Man-linked GlcNAc in N-glycan core structure contributes greatly to the binding. The effect of this GlcNAc is so strong that it can substantially offset the negative effect of substitution on the nonreducing terminal Man residues. On the other hand, the GlcNAc that is usually attached to Asn in N-glycans and the l-Fuc linked at the 6-position of the GlcNAc are irrelevant to the binding. A bisecting GlcNAc neither contributes to nor interferes with the binding. This unique binding specificity of CSL offers many possibilities of its use in analytical and preparative applications. PMID- 10837484 TI - Preparation of completely 6-O-desulfated heparin and its ability to enhance activity of basic fibroblast growth factor. AB - Although regioselective removal of 6-O-sulfate groups of heparin has been undertaken by several researchers, complete 6-O-desulfation with little side reaction has not been attained successfully. In this work, a modified method with a certain silylating reagent, N-methyl-N-(trimethylsilyl)trifluoroacetamide, has been established to produce completely 6-O-desulfated heparin with few other chemical changes. The degrees of 6-O-desulfation were estimated by means of chemical disaccharide analyses and/or (13)C NMR spectra. Although the completely 6-O-desulfated heparin lost about 20% of 2-O-sulfate groups, any other chemical changes and depolymerization were not detected. The completely 6-O-desulfated heparin displayed strong inhibition of COS-1 cell adhesion to basic fibroblast growth factor (bFGF)-coated well in a dose-dependent manner, as was clarified by the competitive cell-adhesion assay. Furthermore, the completely 6-O-desulfated heparin was shown to promote in vitro A31 fibroblast proliferation in a dose dependent manner in the presence of bFGF. These results suggest that signal transduction through bFGF/bFGF receptor in A31 cells occurs in the absence of 6-O sulfate groups in heparin. The involvement of 6-O-sulfate group(s) of heparin/heparan sulfate in the promotion of bFGF mitogenic activity was reported by several groups. This discrepancy between our results and those of other groups would be due to the differences in molecular size of heparin/heparan sulfate derivatives and/or cell species used for the assay. PMID- 10837485 TI - Recombinant fibrinogen studies reveal that thrombin specificity dictates order of fibrinopeptide release. AB - During cleavage of fibrinogen by thrombin, fibrinopeptide A (FpA) release precedes fibrinopeptide B (FpB) release. To examine the basis for this ordered release, we synthesized A'beta fibrinogen, replacing FpB with a fibrinopeptide A like peptide, FpA' (G14V). Analyses of fibrinopeptide release from A'beta fibrinogen showed that FpA release and FpA' release were similar; the release of either peptide followed simple first-order kinetics. Specificity constants for FpA and FpA' were similar, demonstrating that these peptides are equally competitive substrates for thrombin. In the presence of Gly-Pro-Arg-Pro, an inhibitor of fibrin polymerization, the rate of FpB release from normal fibrinogen was reduced 3-fold, consistent with previous data; in contrast, the rate of FpA' release from A'beta fibrinogen was unaffected. Thus, with A'beta fibrinogen, fibrinopeptide release from the beta chain is similar to fibrinopeptide release from the alpha chain. We conclude that the ordered release of fibrinopeptides is dictated by the specificity of thrombin for its substrates. We analyzed polymerization, following changes in turbidity, and found that polymerization of A'beta fibrinogen was similar to that of normal fibrinogen. We analyzed clot structure by scanning electron microscopy and found that clots from A'beta fibrinogen were similar to clots from normal fibrinogen. We conclude that premature release of the fibrinopeptide from the N terminus of the beta chain does not affect polymerization of fibrinogen. PMID- 10837486 TI - BAD Ser-155 phosphorylation regulates BAD/Bcl-XL interaction and cell survival. AB - The BH3 domain of BAD mediates its death-promoting activities via heterodimerization to the Bcl-XL family of death regulators. Growth and survival factors inhibit the death-promoting activity of BAD by stimulating phosphorylation at multiple sites including Ser-112 and Ser-136. Phosphorylation at these sites promotes binding of BAD to 14-3-3 proteins, sequestering BAD away from the mitochondrial membrane where it dimerizes with Bcl-XL to exert its killing effects. We report here that the phosphorylation of BAD at Ser-155 within the BH3 domain is a second phosphorylation-dependent mechanism that inhibits the death-promoting activity of BAD. Protein kinase A, RSK1, and survival factor signaling stimulate phosphorylation of BAD at Ser-155, blocking the binding of BAD to Bcl-XL. RSK1 phosphorylates BAD at both Ser-112 and Ser-155 and rescues BAD-mediated cell death in a manner dependent upon phosphorylation at both sites. PMID- 10837487 TI - Protease-activated receptors 1 and 4 are shut off with distinct kinetics after activation by thrombin. AB - Protease-activated receptors 1 and 4 (PAR1 and PAR4) mediate thrombin signaling in human platelets. Whether these receptors are redundant, interact, or serve only partially overlapping functions is unknown. We report that PAR1 and PAR4 signal with distinct tempos. In transfected fibroblasts, PAR4 triggered substantially more phosphoinositide hydrolysis per activated receptor than PAR1 and was shut off more slowly than PAR1. Shutoff and internalization of PAR1 depends upon phosphorylation of its carboxyl tail upon receptor activation. In contrast to PAR1, phosphorylation of PAR4 was undetectable, and activation dependent internalization of PAR4 was much slower than that seen for PAR1. Mutation of potential phosphorylation sites in the carboxyl tail of PAR1 enhanced PAR1 signaling, whereas analogous mutations in PAR4 had no effect. Thus PAR4 signaling is shut off less rapidly than PAR1, probably due to differences in receptor phosphorylation. PAR1 and PAR4 also signaled with distinct tempos in platelets. PAR1 triggered a rapid and transient increase in intracellular calcium, whereas PAR4 triggered a more prolonged response. Together, the tempo of these responses accounted for that triggered by thrombin. Thus differences in the rates at which PAR1 and PAR4 are shut off allow thrombin to trigger intracellular signaling with distinct temporal characteristics. PMID- 10837488 TI - Plasticity in the primary binding site of galactose/N-acetylgalactosamine specific lectins. Implication of the C-H...O hydrogen bond at the specificity determining C-4 locus of the saccharide in 4-methoxygalactose recognition by jacalin and winged bean (basic) agglutinin I. AB - It is currently believed that an unsubstituted axial hydroxyl at the specificity determining C-4 locus of galactose is indispensable for recognition by galactose/N-acetylgalactosamine-specific lectins. Titration calorimetry demonstrates that 4-methoxygalactose retains binding allegiance to the Moraceae lectin jacalin and the Leguminosae lectin, winged bean (basic) agglutinin (WBA I). The binding reactions were driven by dominant favorable enthalpic contributions and exhibited significant enthalpy-entropy compensation. Proton NMR titration of 4-methoxygalactose with jacalin and WBA I resulted in broadening of the sugar resonances without any change in chemical shift. The alpha- and beta anomers of 4-methoxygalactose were found to be in slow exchange with free and lectin-bound states. Both the anomers experience magnetically equivalent environments at the respective binding sites. The binding constants derived from the dependence of NMR line widths on 4-methoxygalactose concentration agreed well with those obtained from titration calorimetry. The results unequivocally demonstrate that the loci corresponding to the axially oriented C-4 hydroxyl group of galactose within the primary binding site of these lectins exhibit plasticity. These analyses suggest, for the first time, the existence of C-H.O type hydrogen-bond(s) in protein-carbohydrate interactions in general and between the C-4 locus of galactose derivative and the lectins jacalin and WBA I in particular. PMID- 10837489 TI - MCL-1S, a splicing variant of the antiapoptotic BCL-2 family member MCL-1, encodes a proapoptotic protein possessing only the BH3 domain. AB - MCL-1 (myeloid cell leukemia-1) is an antiapoptotic BCL-2 family protein discovered as an early induction gene during myeloblastic leukemia cell differentiation. This survival protein has the BCL-2 homology (BH) domains 1, 2, and 3 and a C-terminal transmembrane region. We identified a short splicing variant of the MCL-1 mRNA in the human placenta encoding a protein, termed MCL-1 short (MCL-1S), with an altered C terminus as compared with the full-length MCL-1 long (MCL-1L), leading to the loss of BH1, BH2, and the transmembrane domains. Analysis of the human MCL-1 gene indicated that MCL-1S results from the splicing out of exon 2 during mRNA processing. MCL-1S, unlike MCL-1L, does not interact with diverse proapoptotic BCL-2-related proteins in the yeast two-hybrid system. In contrast, MCL-1S dimerizes with MCL-1L in the yeast assay and coprecipitates with MCL-1L in transfected mammalian cells. Overexpression of MCL-1S induces apoptosis in transfected Chinese hamster ovary cells, and the MCL-1S action was antagonized by the antiapoptotic MCL-1L. Thus, the naturally occurring MCL-1S variant represents a new proapoptotic BH3 domain-only protein capable of dimerizing with the antiapoptotic MCL-1L. The fate of MCL-1-expressing cells could be regulated through alternative splicing mechanisms and interactions of the resulting anti- and proapoptotic gene products. PMID- 10837491 TI - Accumulation of GTP-bound RhoA during cytokinesis and a critical role of ECT2 in this accumulation. AB - We developed a new pull-down assay for GTP-Rho and examined its level during cell cycle. HeLa cells were arrested in the S phase by thymidine and were enriched in the prometaphase, metaphase, telophase, and G(1) phase by collecting at 0, 45, 90, and 180 min after the release from the nocodazole arrest, respectively. The level of GTP-Rho did not change significantly from the S phase to the prometaphase, but increased thereafter, peaking in the telophase, and returned to the original level in the G(1) phase. The GDP-GTP exchange activity for Rho measured in cell lysates in parallel increased also during the mitosis with a peak in the metaphase. Using this system, we examined a role of ECT2, an exchanger for Rho GTPases, suggested to be involved in cytokinesis (Tatsumoto, T., Xie, X., Blumenthal, R., Okamoto, I., and Miki., T. (1999) J. Cell. Biol. , 147, 921-928). Expression of the dominant negative form of ECT2 completely suppressed both the rise of GTP-Rho in the telophase and the increased GDP-GTP exchange activity in the mitotic cell extracts. These results suggest a critical role of ECT2 in Rho activation during cytokinesis. PMID- 10837490 TI - Novel gain-of-function mutations of platelet glycoprotein IBalpha by valine mutagenesis in the Cys209-Cys248 disulfide loop. Functional analysis under statis and dynamic conditions. AB - Platelet-type von Willebrand disease is a bleeding disorder resulting from gain of-function mutations of glycoprotein (GP) Ibalpha that increase its affinity for von Willebrand factor (vWf). The two known naturally occurring mutations, G233V and M239V, both enrich the valine content of an already valine-rich region within the Cys(209)-Cys(248) disulfide loop. We tested the effect of converting other non-valine residues in this region to valine. Of 10 mutants expressed in CHO cells as components of GP Ib-IX complexes, four displayed a gain-of-function phenotype (G233V, D235V, K237V, and M239V) based on (125)I-vWf binding and adhesion to immobilized vWf. The remainder displayed loss-of-function phenotypes. The gain-of-function mutants bound vWf spontaneously and had a heightened response to low concentrations of ristocetin or botrocetin, whereas the loss-of function mutants bound vWf more poorly than wild-type GP Ibalpha. No distinct gain- or loss-of-function conformations were identified with conformation sensitive antibodies. Compared with cells expressing wild-type GP Ibalpha, cells expressing the gain-of-function mutants rolled significantly more slowly over immobilized vWf under flow than wild-type cells and were able to adhere to vWf coated at lower densities. In aggregate, these data indicate that the region of GP Ibalpha bounded by Asn(226) and Ala(244) regulates the affinity for vWf. PMID- 10837492 TI - Receptor-mediated regulation of the nonselective cation channels TRPC4 and TRPC5. AB - Mammalian transient receptor potential channels (TRPCs) form a family of Ca(2+) permeable cation channels currently consisting of seven members, TRPC1-TRPC7. These channels have been proposed to be molecular correlates for capacitative Ca(2+) entry channels. There are only a few studies on the regulation and properties of the subfamily consisting of TRPC4 and TRPC5, and there are contradictory reports concerning the possible role of intracellular Ca(2+) store depletion in channel activation. We therefore investigated the regulatory and biophysical properties of murine TRPC4 and TRPC5 (mTRPC4/5) heterologously expressed in human embryonic kidney cells. Activation of G(q/11)-coupled receptors or receptor tyrosine kinases induced Mn(2+) entry in fura-2-loaded mTRPC4/5-expressing cells. Accordingly, in whole-cell recordings, stimulation of G(q/11)-coupled receptors evoked large, nonselective cation currents, an effect mimicked by infusion of guanosine 5'-3-O-(thio)triphosphate (GTPgammaS). However, depletion of intracellular Ca(2+) stores failed to activate mTRPC4/5. In inside out patches, single channels with conductances of 42 and 66 picosiemens at -60 mV for mTRPC4 and mTRPC5, respectively, were stimulated by GTPgammaS in a membrane confined manner. Thus, mTRPC4 and mTRPC5 form nonselective cation channels that integrate signaling pathways from G-protein-coupled receptors and receptor tyrosine kinases independently of store depletion. Furthermore, the biophysical properties of mTRPC4/5 are inconsistent with those of I(CRAC), the most extensively characterized store-operated current. PMID- 10837493 TI - MTABC3, a novel mitochondrial ATP-binding cassette protein involved in iron homeostasis. AB - Atm1p, a mitochondrial half-type ATP-binding cassette (ABC) protein in Saccharomyces cerevisiae, transports a precursor of the iron-sulfur (Fe/S) cluster from mitochondria to the cytosol. We have identified a novel half-type human ABC protein, designating it MTABC3 (mammalian mitochondrial ABC protein 3). MTABC3 mRNA is ubiquitously expressed in all of the rat and human tissues examined. MTABC3 protein is shown to be present in the mitochondria, as assessed by immunoblot analysis and confocal microscopic analysis of subcellular fractions of Chinese hamster ovary cells stably expressing MTABC3. Accumulation of iron in the mitochondria, mitochondrial DNA damage, and respiratory dysfunction in the yeast ATM1 mutant strain (atm1-1 mutant cells) were almost fully reversed by expressing MTABC3 in these mutant cells. These results indicate that MTABC3 is a novel ortholog of the yeast and suggest an important role in mitochondrial function. Interestingly, the human MTABC3 gene has been mapped to chromosome 2q36, a region within the candidate locus for lethal neonatal metabolic syndrome, a disorder of the mitochondrial function associated with iron metabolism, indicating that MTABC3 is a candidate gene for this disorder. PMID- 10837494 TI - Evidence against functional heteromultimerization of the KATP channel subunits Kir6.1 and Kir6.2. AB - K(ATP) channels consist of pore-forming potassium inward rectifier (Kir6.x) subunits and sulfonylurea receptors (SURs). Although Kir6.1 or Kir6.2 coassemble with different SUR isoforms to form heteromultimeric functional K(ATP) channels, it is not known whether Kir6.1 and Kir6.2 coassemble with each other. To define the molecular identity of K(ATP) channels, we used adenoviral gene transfer to express wild-type and dominant-negative constructs of Kir6.1 and Kir6.2 in a heterologous expression system (A549 cells) and in native cells (rabbit ventricular myocytes). Dominant-negative (DN) Kir6.2 gene transfer suppressed current through heterologously expressed SUR2A + Kir6.2 channels. Conversely, DN Kir6.1 suppressed SUR2B + Kir6.1 current but had no effect on coexpressed SUR2A + Kir6. 2. We next probed the ability of Kir6.1 and Kir6.2 to affect endogenous K(ATP) channels in adult rabbit ventricular myocytes, using adenoviral vectors to achieve efficient gene transfer. Infection with the DN Kir6.2 virus for 72 h suppressed pinacidil-inducible K(ATP) current density measured by whole-cell patch clamp. However, there was no effect of infection with the DN Kir6.1 on the K(ATP) current. Based on these functional assays, we conclude that Kir6.1 and Kir6.2 do not heteromultimerize with each other and that Kir6.2 is the sole K(ATP) pore-forming subunit in the surface membrane of heart cells. PMID- 10837495 TI - Dynamic interaction of plastocyanin with the cytochrome bf complex. AB - The interaction between plastocyanin and the intact cytochrome bf complex, both from spinach, has been studied by stopped-flow kinetics with mutant plastocyanin to elucidate the site of electron transfer and the docking regions of the molecule. Mutation of Tyr-83 to Arg or Leu provides no evidence for a second electron transfer path via Tyr-83 of plastocyanin, which has been proposed to be the site of electron transfer from cytochrome f. The data found with mutations of acidic residues indicate that both conserved negative patches are essential for the binding of plastocyanin to the intact cytochrome bf complex. Replacing Ala-90 and Gly-10 at the flat hydrophobic surface of plastocyanin by larger residues slowed down and accelerated, respectively, the rate of electron transfer as compared with wild-type plastocyanin. These opposing effects reveal that the hydrophobic region around the electron transfer site at His-87 is divided up into two regions, of which only that with Ala-90 contributes to the attachment to the cytochrome bf complex. These binding sites of plastocyanin are substantially different from those interacting with photosystem I. It appears that each of the two binding regions of plastocyanin is split into halves, which are used in different combinations in the molecular recognition at the two membrane complexes. PMID- 10837496 TI - Identification of an Fe protein residue (Glu146) of Azotobacter vinelandii nitrogenase that is specifically involved in FeMo cofactor insertion. AB - The Fe protein of nitrogenase has three separate functions. Much is known about the regions of the protein that are critical to its function as an electron donor to the MoFe protein, but almost nothing is known about the regions of the protein that are critical to its functions in either FeMo cofactor biosynthesis or FeMo cofactor insertion. Using computer modeling and information obtained from Fe protein mutants that were made decades ago by chemical mutagenesis, we targeted a surface residue Glu(146) as potentially being involved in FeMo cofactor biosynthesis and/or insertion. The Azotobacter vinelandii strain expressing an E146D Fe protein variant grows at approximately 50% of the wild type rate. The purified E146D Fe protein is fully functional as an electron donor to the MoFe protein, but the MoFe protein synthesized by that strain is partially ( approximately 50%) FeMo cofactor-deficient. The E146D Fe protein is fully functional in an in vitro FeMo cofactor biosynthesis assay, and the strain expressing this protein accumulates "free" FeMo cofactor. Assays that compared the ability of wild type and E146D Fe proteins to participate in FeMo cofactor insertion demonstrate, however, that the mutant is severely altered in this last reaction. This is the first known mutation that only influences the insertion reaction. PMID- 10837497 TI - Class A scavenger receptor up-regulation in smooth muscle cells by oxidized low density lipoprotein. Enhancement by calcium flux and concurrent cyclooxygenase-2 up-regulation. AB - Oxidative stress caused by phorbol esters or reactive oxygen up-regulates the class A scavenger receptor (SR-A) in human smooth muscle cells (SMC), which normally do not express this receptor. The increase in SR-A expression correlates with activation of the redox-sensitive transcription factors activating protein-1 c-Jun and CCAAT enhancer-binding protein beta. Here we show that coincubation of SMC with macrophages or oxidized low density lipoproteins (LDL) from macrophage conditioned medium activates these same regulatory pathways and stimulates SR-A expression. The increased SR-A gene transcription induced by cell-oxidized LDL up regulated SR-A mRNA and increased by 30-fold the uptake of acetyl LDL, a ligand for the SR-A. Copper-oxidized LDL also increased SR-A receptor expression. Oxidized LDL with a lipid peroxide level of 80-100 nmol/mg of LDL protein and an electrophoretic mobility approximately 1.5 times that of native LDL exhibited the greatest bioactivity. Inhibition of calcium flux suppressed SR-A induction by oxidized LDL. Conversely, calcium ionophore greatly enhanced SR-A up-regulation by oxidized LDL or other treatments that promote intracellular oxidative stress. This enhancement was dependent upon concurrent up-regulation of SMC cyclooxygenase-2 expression and activity and was blocked by the cyclooxygenase-2 inhibitors NS-398 and Resveratrol. In THP-1 cells, oxidized LDL induced monocyte to-macrophage differentiation and increased SR-A expression. These findings support a role for mildly oxidized LDL in the redox regulation of macrophage differentiation and SR-A expression and suggest that increased vascular oxidative stress may contribute to the formation of both SMC and macrophage foam cells. PMID- 10837498 TI - Molecular mechanisms of tumor necrosis factor alpha gene expression in monocytic cells via hyperglycemia-induced oxidant stress-dependent and -independent pathways. AB - Increased oxidative stress has been reported in vivo in the diabetic state via the production of reactive oxygen species (ROS). Such stress is bound to play a key role on activation of circulating monocytes, leading to the accelerated atherosclerosis observed in diabetics. However the exact molecular mechanisms of monocyte activation by high glucose is currently unclear. Here, we demonstrate that chronic high glucose (CHG) causes a dramatic increase in the release of the inflammatory cytokine tumor necrosis factor alpha (TNFalpha), at least in part through enhanced TNFalpha mRNA transcription, mediated by ROS via activation of transcription factors nuclear factor kappaB (NF-kappaB) and activating protein-1 (AP-1). TNFalpha accumulation in the conditioned media was increased 10-fold and mRNA levels were increased 11.5-fold by CHG. The following observations supported that both NF-kappaB and AP-1 mediated enhanced TNFalpha transcription by CHG: 1) A 295-base pair fragment of the proximal TNFalpha promoter containing NF-kappaB and AP-1 sites reproduced the effects of CHG on TNFalpha transcription in a luciferase reporter assay, 2) mutational analyses of both NF-kappaB and the AP-1 sites abrogated 90% of the luciferase activity, 3) gel-shift analysis using the binding sites showed activation of NF-kappaB and AP-1 in CHG nuclear extracts, and 4) Western blot analyses demonstrated elevated nuclear levels of p65 and p50 and decreased cytosolic levels of IkappaBalpha in CHG-treated monocytes. That ROS acted as a key intermediate in the CHG pathway was supported by the following evidence: 1) increased superoxide levels similar to those observed with PMA or TNFalpha, 2) increased phosphorylation of stress-responsive mitogen-activated protein kinases p38 and JNK-1, 3) counteraction of the effects of CHG on TNFalpha production, the 295TNFluc reporter activity, activation of NF-kappaB, and repression of IkappaBalpha by antioxidants and p38 mitogen-activated protein kinase inhibitors. The study suggests that ROS function as key components in the regulatory pathway progressing from elevated glucose to monocyte activation. PMID- 10837499 TI - Sialomucin complex (rat Muc4) is regulated by transforming growth factor beta in mammary gland by a novel post-translational mechanism. AB - Sialomucin complex (SMC, rat Muc4) is a heterodimeric glycoprotein complex consisting of a mucin subunit ASGP-1 (for ascites sialoglycoprotein-1) and a transmembrane subunit ASGP-2, produced from a single gene and precursor. SMC expression is tightly regulated in mammary gland; the level in lactating mammary gland is about 100-fold that in virgin gland. In rat mammary epithelial cells, SMC is post-transcriptionally regulated by Matrigel by inhibition of SMC precursor synthesis. SMC is also post-transcriptionally regulated by transforming growth factor-beta (TGFbeta). The repression of SMC expression by TGFbeta is rapid, is independent of TGFbeta-induced cell cycle arrest, and does not require new protein synthesis. Unlike Matrigel, TGFbeta does not reduce SMC protein synthesis, as SMC precursor accumulation is equivalent in TGFbeta-treated and untreated cells. Instead, SMC precursor in TGFbeta-treated cells is more persistent and does not become processed as rapidly into mature ASGP-1 and ASGP 2, indicating that TGFbeta disrupts processing of SMC precursor. These results indicate that SMC, a product of normal mammary gland and milk, is regulated by TGFbeta by a novel post-translational mechanism. Thus, SMC is regulated by multiple post-transcriptional mechanisms, which serve to repress potential deleterious effects of overexpression. PMID- 10837500 TI - Introducing the model systems series PMID- 10837501 TI - A systems approach to the cellular analysis of associative learning in the pond snail Lymnaea. AB - We show that appetitive and aversive conditioning can be analyzed at the cellular level in the well-described neural circuitries underlying rhythmic feeding and respiration in the pond snail, Lymnaea stagnalis. To relate electrical changes directly to behavior, the snails were first trained and the neural changes recorded at multiple sites in reduced preparations made from the same animals. Changes in neural activity following conditioning could be recorded at the level of motoneurons, central pattern generator interneurons and modulatory neurons. Of significant interest was recent work showing that neural correlates of long-term memory could be recorded in the feeding network following single-trial appetitive chemical conditioning. Available information on the synaptic connectivity and transmitter content of identified neurons within the Lymnaea circuits will allow further work on the synaptic and molecular mechanisms of learning and memory. PMID- 10837502 TI - Cellular and molecular mechanisms underlying learning and memory impairments produced by cannabinoids. AB - Why does smoking marijuana impair learning and memory? Behavioral studies suggest that a disruption of normal hippocampal function contributes to these deficits. In vitro experiments find that cannabinoid receptor activation reduces neurotransmitter release below the levels required to trigger long-term changes in synaptic strength in the hippocampus. Cannabinoids reduce glutamate release through a G-protein-mediated inhibition of the calcium channels responsible for neurotransmitter release from hippocampal neurons. These mechanisms likely play a role in the learning and memory impairments produced by cannabinoids and by endogenous cannabinoid receptor ligands. PMID- 10837503 TI - Operant conditioning in Lymnaea: evidence for intermediate- and long-term memory. AB - Aerial respiration of the pond snail, Lymnaea stagnalis, can be operantly conditioned; however, the parameters necessary to produce long-term (LTM) or intermediate term memory (ITM) have not previously been investigated. We conducted training using procedures that varied in the duration of the training session, the number of training sessions per day or the amount of time between subsequent training sessions (SI). We found that by varying the duration and frequency of the training session learning could be differentially produced. Furthermore, the ability to form LTM was dependent not only on the duration of the training session was also the interval between training sessions, the SI. Thus it was possible to produce ITM, which persists for up to 3 hr, and not form LTM, which persists at least 18 hr. Learning, ITM, and LTM can be differentially produced by altering the SI, the duration of the training session, or the number of training sessions per day. These findings may allow us to begin to elucidate the underlying neural mechanisms of learning, ITM, and LTM. PMID- 10837504 TI - Contextual modulation of memory consolidation. AB - We investigate olfactory memory consolidation in honeybees. Three experiments are reported that include 1024 animals in 28 experimental groups. After one pairing of odorant and sucrose reward, retention is typically nonmonotonic with a minimum 3 min after conditioning. This corresponds to the "Kamin effect" in vertebrates; the postminimum rise in retention is usually interpreted as reflecting memory consolidation. First, we test for the generality of this effect across four different odorants. The postminimum rise in retention was reproducibly observed for 1-hexanol but not for 1-octanol, limonene, or geraniol. Second, we investigate whether previous learning about the training context modulates subsequent memory consolidation. On the day before training, a reward was applied either upon placement into the future training context for 1 min, halfway during exposure or just before removal from the context. In the latter group, the 3-min minimum in retention was eliminated; thus, in that group, forward pairings of context and reward (i.e., context exposure begins before reward is applied) lead to an associative context memory that can modulate subsequent olfactory memory consolidation. Third, we found no evidence for a modulation of olfactory memory consolidation by pre-exposure to the odorant. PMID- 10837505 TI - Further evidence for a dissociation between different forms of mnemonic expressions in a mouse model of age-related cognitive decline: effects of tacrine and S 17092, a novel prolyl endopeptidase inhibitor. AB - It has been demonstrated previously on the radial maze that the emergence of an age-related mnemonic impairment is critically dependent on the form which the discrimination problems took. Hence, when the arms were presented one by one (i.e., successive go-no-go discrimination), both adult and aged mice learned to distinguish between positive (baited) and negative (unbaited) arms readily, as evidenced by their increased readiness to enter positive relative to negative arms (i.e., by a differential in arm-entry latencies). A selective impairment in the aged mice was seen when these arms were presented subsequently as pairs, such that the mice were confronted with an explicit choice (i.e., simultaneous 2 choice discrimination). When discriminative performance was measured by the differential run speed between positive and negative arms, aged mice were also impaired. This was particularly pronounced in the 2-choice discrimination condition. We examined the effects of tacrine (3mg/kg, subcutaneously) or S 17092 (10mg/kg, orally) in aged mice on the three behavioral indices of this 2-stage spatial discrimination paradigm. The results indicated that: (1) Tacrine, but not S 17092, enhanced the acquisition of go-no-go discrimination as reflected in arm entry latencies; (2) both drugs improved choice accuracy in simultaneous discrimination, although the effect of tacrine was less striking and, in particular, far from statistical significance in the very first 2-choice responses; and (3) neither drugs significantly affected run-speed performance. We conclude further that the specific patterns of drug effects on the three indices of discriminative performance might suggest that each index is associated with a distinct form of mnemonic expression relying on separate neural systems. PMID- 10837507 TI - Impaired emotional declarative memory following unilateral amygdala damage. AB - Case studies of patients with bilateral amygdala damage and functional imaging studies of normal individuals have demonstrated that the amygdala plays a critical role in encoding emotionally arousing stimuli into long-term declarative memory. However, several issues remain poorly understood: the separate roles of left and right amygdala, the time course over which the amygdala participates in memory consolidation, and the type of knowledge structures it helps consolidate. We investigated these questions in eight subjects with unilateral amygdala damage, using several different measures. For comparison, our main task used stimuli identical to those used previously to investigate emotional declarative memory in patients with bilateral amygdala damage. Contrasts with both brain damaged and normal control groups showed that subjects with left amygdala damage were impaired in their memory for emotional stimuli, despite entirely normal memory for neutral stimuli (because of a number of caveats, the findings from subjects with right amygdala damage were less clear). Follow-up experiments suggested that the normal facilitation of memory for emotional stimuli may develop over an extended time course (>30 min), consistent with prior findings, and that the specific impairment we report may depend in part on the lexical nature of the task used (written questionnaire). We stress the complex and temporally extended nature of memory consolidation and suggest that the amygdala may influence specific components of this process. PMID- 10837506 TI - Strain-dependent differences in LTP and hippocampus-dependent memory in inbred mice. AB - Many studies have used "reverse" genetics to produce "knock-out" and transgenic mice to explore the roles of various molecules in long-term potentiation (LTP) and spatial memory. The existence of a variety of inbred strains of mice provides an additional way of exploring the genetic bases of learning and memory. We examined behavioral memory and LTP expression in area CA1 of hippocampal slices prepared from four different inbred strains of mice: C57BL/6J, CBA/J, DBA/2J, and 129/SvEms-+(Ter?)/J. We found that LTP induced by four 100-Hz trains of stimulation was robust and long-lasting in C57BL/6J and DBA/2J mice but decayed in CBA/J and 129/SvEms-+(Ter?)/J mice. LTP induced by one 100-Hz train was significantly smaller after 1 hr in the 129/SvEms-+(Ter?)/J mice than in the other three strains. Theta-burst LTP was shorter lasting in CBA/J, DBA/2J, and 129/SvEms-+(Ter?)/J mice than in C57BL/6J mice. We also observed specific memory deficits, among particular mouse strains, in spatial and nonspatial tests of hippocampus-dependent memory. CBA/J mice showed defective learning in the Morris water maze, and both DBA/2J and CBA/J strains displayed deficient long-term memory in contextual and cued fear conditioning tests. Our findings provide strong support for a genetic basis for some forms of synaptic plasticity that are linked to behavioral long-term memory and suggest that genetic background can influence the electrophysiological and behavioral phenotypes observed in genetically modified mice generated for elucidating the molecular bases of learning, memory, and LTP. PMID- 10837508 TI - D2 dopamine receptor blockade immediately post-training enhances retention in hidden and visible platform versions of the water maze. AB - Considerable evidence shows that post-training administration of dopamine agonists can enhance memory through actions on consolidation processes, but relatively little is known regarding the effects of dopamine antagonists on consolidation. These experiments investigated the effects of post-training systemic administration of the D2 receptor antagonist sulpiride on consolidation of memory for two versions of the Morris water maze task. Rats trained in either the hidden (spatial) or visible (cued) platform version received a subcutaneous injection of sulpiride or vehicle immediately following training. Retention testing 48 hr later revealed that relative to vehicle controls, sulpiride reduced platform latencies in both task versions, suggesting that like dopamine agonists, sulpiride can also have memory-enhancing effects. PMID- 10837509 TI - Cloning, modeling, and chromosomal localization for a small leucine-rich repeat proteoglycan (SLRP) family member expressed in human eye. AB - PURPOSE: To examine a highly abundant novel transcript from human iris. METHODS: Expressed sequence tag (EST) analysis of an adult human iris cDNA library revealed an abundant (>0.7%) transcript for a novel member of the small leucine rich proteoglycan (SLRP) family. Other 3' ESTs from retina were also detected in dbEST. The structure of the leucine-rich repeat (LRR) domain was investigated by molecular modeling. Antisera were raised against a specific peptide and used in western blots of human and rat eye tissues. RESULTS: From its prevalence in the eye and its superfamily relationships, this SLRP protein has been given the names oculoglycan or opticin (Optc). Sequence analysis suggests that Optc has a signal peptide and two structural domains, the larger of which is the LRR domain. Modeling of the LRR domain reveals structural variability in the repeat motifs, forming potential interaction sites for binding partners. Antiserum to a specific peptide detected a protein of approximately 48 kDa, in human iris, ciliary body and retina while the major protein detected in rat ocular tissues was 37 kDa in size. This may reflect a species difference in post-translational modification. Radiation hybrid mapping shows that the gene for OPTC is located on chromosome 1q31, close to the inherited eye diseases ARMD1 and AXPC1. CONCLUSIONS: Optc is a newly identified SLRP family member, which appears to have eye-preferred expression. Molecular modeling reveals local deviations from the familiar LRR structure, which are candidates for specific interaction sites. Western blotting with a specific peptide antibody detects Optc in iris, ciliary body and retina in the human eye and suggests that the protein is post-translationally modified. In rat, the antibody detects Optc in several eye tissues and in brain but the protein appears to have undergone much less modification, suggesting that this is not essential for all aspects of function. Considering its eye-preferred expression, the OPTC gene has the potential for involvement in inherited eye disease. Indeed, it maps close to at least two disease loci for which no gene has so far been identified. PMID- 10837510 TI - The human gene for gammaS-crystallin: alternative transcripts and expressed sequences from the first intron. AB - PURPOSE: gammaS-crystallins are major components of adult vertebrate lenses. Here we examine the population of gammaS transcripts in adult human lens and the structure of the human CRYGS genes. METHODS: Adult lens human transcripts were obtained from NEIBANK, an Expressed Sequence Tag (EST) analysis of human eye tissues. The human CRYGS gene was isolated as a PAC clone and sequenced by direct and PCR-based methods. RESULTS: As judged by EST frequency, gammaS is one of the most abundant transcripts in the adult human lens, ranking just behind betaB2-, alphaB- and alphaA-crystallins. EST analysis reveals two transcript sizes resulting from alternative AATAAA and ATTAAA polyadenylation signals. In addition, one cDNA clone was found to contain a novel insert sequence that disrupted the open reading frame. Gene sequencing confirmed that this insert comes from intron 1 and is part of a sequence corresponding to a cluster of unidentified human transcripts in dbEST. Human and mouse gammaS gene proximal promoter sequences were compared and showed a high degree of evolutionary conservation, including consensus binding sites for transcription factors of the maf and SOX families. CONCLUSIONS: The human CRYGS gene can give rise to at least two transcripts through alternative polyadenylation. A minor transcript results from alternative splicing into sequences in intron 1. These sequences form part of a transcription unit (Mys) expressed in several non-lens tissues. The identity and function Mys of is not yet known, however, the cryptic splicing of CRYGS could produce a defective protein product, with potentially deleterious results for the adult human lens. PMID- 10837511 TI - Temperature-modulated high-performance liquid chromatography for detecting variation in human genome. AB - Recently, temperature modulated heteroduplex analysis (TMHA) based upon ion-pair reversed-phase high-performance liquid chromatography has been developed as a tool for polymorphisms and novel mutation detection in human genome. This new approach, with increased accuracy and throughput over traditional methods. This new approach is also known as denaturing high performance liquid chromatography(DHPLC). However, the terms of both TMHA and DHPLC should be considered carefully. Firstly, it is DNA molecular that is denatured in high performance liquid chromatography but not the method or the instrument. Secondly, the temperature in the liquid chromatograph is modulated according to Tm (melting temperature) of the DNA fragments. As it is known, Tm is the temperature at which a population of double-stranded nucleic acid molecules becomes half-dissociated into single strands, and under this temperature, half of the DNA molecules is still not denaturing. Thirdly, the targets for detection are both the homoduplexes and the mismatched heteroduplexes, which are double-stranded nucleic acid molecules and are annealing. The term of DHPLC may lead to confusion in annealing and denaturing. In addition, since TMHA can be carried out using gel electrophoresis and HPLC, there is a need to distinguish HPLC from gel electrophoresis. Therefore, we suggest a new term for this technology, which is TmHPLC (temperature-modulated high- performance liquid chromatography). This term not only provides a clear definition for this new method, but also includes both TMHA and HPLC. More importantly, TmHPLC implies that the temperature in HPLC is modulated according to Tm of the DNA fragments. We also review advances in detecting variation through TmHPLC. This new approach leads to significant reduction of sequencing efforts for searching novel mutations and polymorphisms, and because of its high sample throughput, to faster data generation. This implies that TmHPLC is a sensitive, accurate, and cost effective approach to screening sequence variation in human genome. PMID- 10837512 TI - Association analysis of variants in the core promoter region of angiotensinogen gene with essential hypertension in Tibetan population. AB - OBJECTIVE: To detect the variants in the core promoter region of angiotensinogen(AGT) gene, and to analyse the relationship between the AGT gene polymorphisms and essential hypertension in Tibetan population. METHODS: This is a case-control study consisting of 103 essential hypertensive subjects and 82 normotensive controls matched by age and sex. The variants in the AGT gene core promoter region were screened by polymerase chain reaction/single strand conformation polymorphism(PCR/SSCP) and further identified by automated sequencing. The A(-6)G polymorphism was determined in DNA extracted from leucocytes by polymerase chain reaction/restriction fragment length polymorphism (PCR/RFLP). RESULTS: (1) There were two different electrophoresis band patterns in PCR/SSCP analysis. PCR product direct sequencing showed that the two band patterns represented the AA, AC genotypes in the (-20) site of AGT gene respectively. The distribution of A(-20)C genotype was almost identical in essential hypertensive and normotensive groups (P>0.8). The A allele frequency was very high in both groups (control: 0.9175, hypertensive: 0.9124). (2)Distribution of genotype in the (-6) site of AGT gene was much different between the patient group and control group (P<0.005). The frequency of G allele was statistically higher in the patient group than in controls (0.374 vs 0.220, P<0.025). CONCLUSION: Both Tibetan hypertensives and normotensives have higher frequency of A allele in AGT gene (-20) site. The higher frequency of G allele in the AGT gene (-6) site in Tibetan hypertension patients suggests that this allele may be the genetic susceptibility factor in the proceeding of essential hypertension in the Tibetan population. PMID- 10837513 TI - Detailed allelic loss mapping on 7q32 in nasopharyngeal carcinoma. AB - OBJECTIVE: The present research group had found high frequency of loss of heterozygosity(LOH) of D7S500-D7S495 on chromosome band 7q32 in nasopharyngeal carcinoma, this study was conducted to further find the deletion region. METHODS: Higher density of loci on 7q32 in 30 tumors was studied by using microsatellite analysis. RESULTS: The frequency of LOH was 63.3%. The highest frequency of LOH was identified at the loci of D7S500-D7S509-D7S495, among which a common deletion of D7S509 was found. CONCLUSION: A putative tumor suppressor gene may be present around D7S509. PMID- 10837514 TI - A preliminary study of single nucleotide polymorphisms of lipoprotein lipase gene in coronary atherosclerotic heart disease. AB - OBJECTIVE: This study inquired into the relationship between single nucleotide polymorphisms(SNPs) of lipoprotein lipase gene (LPL) and coronary atherosclerotic heart disease(CHD). METHODS: Exon 6 and fragment of intron 6 in LPL gene from 102 cases of CHD and 110 normal subjects were analysed by polymerase chain reaction amplification, temperature-modulated high-performance liquid chromatography (TmHPLC) and DNA sequencing. RESULTS: Three new SNPs (4212t/c, 4509t/c, 4576a/c) were detected in the intron 6 of LPL gene. There was significant difference in frequencies of 4212t/c, and 4576a/c between health group and CHD group(P<0.05). CONCLUSION: This study has accumulated additional data on SNPs in LPL gene and provided new data for exploring the mechanism of CHD. PMID- 10837515 TI - An analysis of two polymorphic points of the 7th intron of human p53 gene. AB - OBJECTIVE: To detect polymorphic points of the 7th intron of human p53 oncogene. METHODS: Polymerase chain reaction (PCR) and double- strand DNA direct sequencing were used to analyse sequence alteration of p53 intron 7. One hundred and five cases of normal human peripheral blood samples with no genetic relation were investigated. RESULTS: There were two polymorphic points in the 7th intron of p53 gene. The first one was localized at 73 base pair (bp) to 3'-end of exon 7; the other one at 93. Three genotypes were found. Twenty-two cases were of type TG, 37 cases were of type CT, and the other 46 cases were of heterozygote. Because the first point was alteration of GGGCCC to GGGTCC or its heterozygote, it had a point alteration of enzyme Apa I. CONCLUSION: There are two polymorphic points in the 7th intron of human p53 gene, which may be of importance to identification of individual genetic relation and to judging of a case in forensic medicine. PMID- 10837516 TI - The effects of recombinant RA538 and antisense c-myc adenovirus on tumor cells and the molecular mechanism concerned. AB - OBJECTIVE: Compare the biological effects of recombinant RA538 and antisense c myc adenovirus on human gastric cancer cell line (SGC7901) and explore the molecular mechanism in vitro and in vivo. METHODS: SGC7901 cells were treated with Ad-RA538, Ad-AS c-myc or Ad-LacZ. MTT, DNA ladder, TUNEL and FCM, RT-PCR, Western blot analysis, the tumorigenicity in nude mice and experimental therapy of the nude mice were used. RESULTS: Ad-RA538 and Ad-ASc-myc could strongly inhibit cell growth and induce apoptosis of SGC7901 cells. The growth of the Ad RA538- and Ad-ASc-myc-infected SGC7901 cells were inhibited by 76.3% and 44.1% respectively. The over expression of RA538 and ASc-myc could down-regulate expression of c-myc, bcl-2 and cyclinD1 gene and up-regulate expression of bax gene, but it could not regulate expression of p53, p16, TGase, and ras gene. The tumorigenicity of Ad-RA538 or Ad-ASc-myc in nude mice showed that three of three mice failed to form tumor, compared with Ad-LacZ and parent SGC7901 cells from 7 to 30 days. Experimental therapy of the nude mice bearing subcutaneous tumor of SGC7901 cells showed that intratumor instillation of Ad-RA538 and Ad-ASc-myc inhibited the growth of the tumors. Ad-RA538- and Ad-ASc-myc-treated tumors were inhibited by 60.7% or 68.9% respectively, compared with the tumor injected with Ad-LacZ and mock. CONCLUSION: The expression of Ad-RA538 and Ad-ASc-myc can inhibit growth and induce apoptosis of gastric cancer cell in vitro and in vivo. RA538 and ASc-myc relate to c-myc, bcl-2, cyclinD1 and bax gene closely and have noticeable biologic effects on gastric cancer cells. PMID- 10837517 TI - Detection of point mutations in promoter and coding sequence of p21 gene in a variety of human malignancies. AB - OBJECTIVE: p21 is a general inhibitor of cyclin-dependent kinases which leads cells through restriction point of cell cycle. Since p21 has been identified to be a mediator of p53 tumor suppressor activity and directly up-regulated by p53, this study was designed to investigate whether mutations in promoter or coding sequence of p21 were associated with development or progress of tumor. METHODS: A 2.4kb DNA fragment, containing essential promoter, was amplified and sequenced. Furthermore, 346 DNA samples from 9 different types of primary tumors were examined for point mutations in promoter and coding sequence of p21 gene by using SSCP analysis and direct sequencing. RESULTS: Heterozygous or homozygous G to A transition at 5bp upstream and C to A transversion at 160bp downstream of p53 binding site were found. A 44bp heterozygous deletion was detected in a bladder cancer, which started at codon 34 and resulted in frameshift and a premature stop codon downstream. The most abundant variants in coding sequence of p21 gene were a C to A transversion at codon 31 which resulted in Ser to Arg and had been identified being polymorphism. In addition, four rare variants in coding sequence were also found. CONCLUSION: The evidence from this study indicates that mutation in either promoter or coding sequence of p21 gene is rare in a variety of human malignancies. PMID- 10837518 TI - A genetic study of the depressive respiratory responses to hypoxia in chronic obstructive pulmonary disease patients with type II respiratory failure. AB - OBJECTIVE: The depression of the ventilatory and P(0.1 )responses to hypoxia and hypercapnia may play an important role in the development of CO(2) retention in the patients with chronic obstructive pulmonary disease (COPD). The purpose of this study was to investigate if there is any linkage relationship between the depressed ventilatory and P(0.1) response phenotypes and the microsatellite markers on chromosome 6q21.1-21.2 among Chinese people. METHODS: P(0.1) and ventilatory responses to hypoxia were measured in six COPD patients with type II respiratory failure, as well as in their 21 normal adult offsprings. Polymerase chain reaction (PCR), denaturing polyacrylamide gel electrophoresis and silver staining techniques were used to study the amplified fragment length polymorphism. Linkage analysis was done with the LINKAGE program. RESULTS: Hypoxia response was low in 10 offsprings, and normal in the other 11 offsprings. Linkage analysis showed the maximum Lod score was 1.74(D6S276, theta = 0.10). CONCLUSION: The depressed hypoxia response might have been influenced by genetic factors. The model was in accord with autosomal dominant inheritance. The disease related gene was linked to D6S276 locus. PMID- 10837519 TI - Genetic epidemiology study of pathological myopia. AB - OBJECTIVE: To explore the genetic epidemiology of pathological myopia (PM), including hereditary and genetic model. METHODS: The simple segregation analysis was done by SEGRANB. The values of segregation ratio p and the proportion of sporadic cases x were estimated. The complex segregation analysis was performed using SAGE-REGD. The genetic model and gene frequency were estimated. The 62 pedigrees with PM were random samples from hospital patients. RESULTS: By simple segregation analysis, the genetic pattern of N*N is autosomal recessive and the frequency of sporadic cases is approximately 65.72%. The genetic pattern of A*N may be autosomal recessive (but autosomal dominant cannot be excluded), the frequency of sporadic cases is approximately 35.14%. By complex segregation analysis, the genetic model of PM is autosomal recessive and the gene frequency is 0.147385. CONCLUSION: PM is compatible with autosomal recessive inheritance (autosomal dominant not excluded ), the sporadic cases are existent and the gene frequency is 0.147385. PMID- 10837520 TI - Correlation of genetic polymorphism of cytochrome P4502D6 with dextromethorphan oxidative metabolism in Chinese. AB - OBJECTIVE: To determine the genetic mechanism of dextromethorphan oxidative polymorphism in Chinese subjects. METHODS: CYP2D6 genotype was analyzed by polymerase chain reaction (PCR) and Hph I restriction fragment length polymorphism (RFLP) techniques in 119 healthy Chinese subjects. RESULTS: The gene frequency of CYP2D6*10B was 58.4%, including 13(10.9%) of homozygous wild type(w/w), 33 (27.7%) of homozygous mutant(m/m), and 73(61.3%) of heterozygous genotypes(m/w). Twenty-nine subjects out of 42 intermediate metabolizers(70%) of dextromethorphan were homozygous for CYP2D6*10B. One poor metabolizer subject also showed an m/m genotype. Besides, 10 Chinese subjects were tested and excluded for the presence of any of the six mutant alleles associated with poor metabolism of CYP2D6 in Caucasians. CONCLUSION: The CYP2D6*10B allele containing the C(188)--> T mutation is the major cause of CYP2D6 polymorphism in relation to diminished dextromethorphan oxidative capacity in Chinese subjects. PMID- 10837521 TI - Genetic heterogeneity of non-specific mental retardation. AB - OBJECTIVE: To explore genetic patterns of non-specific mental retardation (NSMR). METHODS: Using segregation analysis, Finney method and Falconer method, the authors studied 157 pedigrees with NSMR selected in a population survey in Shandong province. RESULTS: Ux U multiplex families may accept the autosomal recessive inheritance, Ux U total families is the autosomal recessive inheritance while it may be the multifactorial inheritance with major-gene effect, the frequency of sporadic cases is approximately 46%; Ux A mating families may keep the incomplete penetrance in autosomal dominant inheritance. CONCLUSION: The genetic pattern of NSMR has the genetic heterogeneity. PMID- 10837522 TI - Linkage of the genes controlling natural killer cell activity to HLA-B. AB - OBJECTIVE: To study whether there is a relationship between HLA-B alleles and low natural killer(NK) cell activity in Chinese. METHODS: The subjects were 34 unrelated healthy individuals (10 males and 24 females), aged 25-60 years, and 31 individuals (17 males and 14 females) from 11 families, aged 22-70 years, previously genotyped for HLA class I, II, complotypes and haplotypes that were assigned based on family studies. The unrelated individuals were typed for HLA-A, -B antigen by a microcytotoxicity assay and all the subjects were tested for NK cell activity by lactate dehydrogenase (LDH) enzyme-release assay. RESULTS: Statistical analysis showed that 30 lysis unit(LU) was the cutoff value of low and high NK reactors. The frequency of low reactor individuals homozygous for HLA B was significantly higher than that for those for HLA-DR(P< 0. 05). The associated genes included HLA-B13, B40, B46, B7, B57, defined as NKB1 genes, and HLA-B44 as NKB2 gene. Family studies provide further evidence that the genes controlling low NK cell activity were located in the MHC region and it was a recessive trait. CONCLUSION: There were recessive genes associated with HLA-B controlling low NK cell activity in Chinese. The NKB alleles were partly different from those found in Caucasian. PMID- 10837523 TI - Study on DNA polymorphism at D1S8 locus in Hebei Han population. AB - OBJECTIVE: To study the polymorphism at D1S8 locus and to provide basic information for the construction of DNA fingerprint database and the application in forensic medicine. METHODS: Minisatellite variant repeat-polymerase chain reaction (MVR-PCR) and polyacrylamide gradient gel electrophoresis followed by silver staining were used to detect the variant repeat sequences at D1S8 locus of 240 unrelated individuals in Hebei Han population, and digital codes were obtained. RESULTS: Each individual obtained about 30 digital codes, in which none of two unrelated individuals had the same code. The probability of identity of 30 digital codes was 3.55x10(-11). The percentages of three repeat units, a-type, t type and o-type were 54.77%, 42.54% and 2.69% respectively. The heterozygosity (H) was 0.9837. The excluding probability of paternity(EPP) and polymorphism information content(PIC) were 0.9669 and 0.9833 respectively. CONCLUSION: Because of its high polymorphic nature, D1S8 locus is a valuable marker for forensic identification and paternity testing. PMID- 10837524 TI - Detection of mosaic chromosome 21 aneuploidy in vivo with CB-FISH method. AB - OBJECTIVE: To explore the mechanism by which the ratio of mosaic diploid cells in vivo increased in trisomy 21 cases. METHODS: Fluorescence in situ hybridization (FISH) with pericentric DNA probes specific to human chromosome 21 to the binucleated cells cytokinesis- blocked by cytochalasin-B(CB), namely, CB-FISH method. RESULTS: The mean frequency of diploid cells (1.69%+/-1.07%) was significantly higher than that of tetrasomy 21 cells(0.042%+/-0.064%)(P=0.000) in 12 trisomy 21 cases aged 0-9 years (mean 4 years) in vivo; whereas the incidence of spontaneous monosomy 21 cells (0. 265%+/-0.13%) was close to the frequency of trisomy 21 cells (0. 225%+/-0.184%) in 20 control individuals aged 0-10 years (mean 5 years). The frequency of cells undergone chromosome 21 nondisjunction was 0.822%+/-0.554% in cultured trisomy 21 cells, which was significantly higher than that in the cultured diploid cells (0.369%+/-0.25%). However, the rates of cells with chromosome 21 loss in the two cultured cell lines (0.01%+/-0.03% vs 0.043%+/ 0.049%) were not significantly different. In both cultured cell lines, chromosome 21 nondisjunction occurred more frequently than loss. CONCLUSION: The increased frequency of diploidies in trisomy 21 cases may result from the accumulation of diploid cells originating from malsegregation of chromosome 21 in trisomic cells rather than the increase of chromosome 21 loss. PMID- 10837525 TI - Genetic polymorphism of 4 STR loci on chromosome 17 in Chinese Han. AB - OBJECTIVE: To analyze genetic polymorphism of D17S1290, D17S1293, D17S1303 and D17S1308 in Chinese Hans. METHODS: Fifty unrelated individuals were analyzed by PCR amplification fragment length polymorphism analysis method. RESULTS: 9, 7, 5, 5 alleles were observed at these 4 STR loci respectively, the genotypes distributions in Chinese Hans were in accordance with Hardy-Weinberg equilibrium, the expected heterozygosities for these loci were 0.770, 0.828, 0.608, 0.669 respectively, the polymorphism information contents(PIC) were 0.763, 0.820, 0.602, 0.662 respectively. CONCLUSION: The results demonstrate these 4 STR loci can be used for genetic analysis. PMID- 10837526 TI - Analysis of temperature-modulated high-performance liquid chromatography as a tool for detection of single nucleotide polymorphism. AB - OBJECTIVE: To establish and evaluate temperature-modulated high-performance liquid chromatography(TmHPLC) as a rapid and efficient technique of detecting single nucleotide polymorphism(SNP). METHODS: Several TmHPLC parameters, such as the eluting gradient ranges and column temperatures used for analysing the amplicons have been studied. The size of DNA fragments suitable for analysis of SNP was studied. Also the sensitivity of the TmHPLC was evaluated. RESULTS: The suitable chromatographic condition of TmHPLC was found out. The elution peaks of dNTP and primers could be separated from those of polymerase chain reaction products of the genomic DNA. The heterozygotes of SNP from the sequenced DNA fragment with 178bp, 269bp and 272bp could be distinguished from the homozygotes of SNP clearly. CONCLUSION: The authors have established and evaluated the TmHPLC method, which is an efficient, sensitive and rapid method for screening out SNP in the DNA fragments of larger size. PMID- 10837527 TI - Preface. PMID- 10837528 TI - Preparation of peptide and protein powders for inhalation. AB - Innovations in biotechnology and recombinant techniques have led to a large increase in the number of macromolecule drugs developed over the last several years. These molecules are enabling new therapies for many previously untreated or poorly treated diseases. Due to poor oral absorption, most biopharmaceuticals are delivered by injection. Medical science, health care providers and consumers have been searching for alternative methods of delivering macromolecule drugs. Animal and human clinical studies have demonstrated that pharmacological doses of bioactive peptides and proteins pass, by a natural biological process, from the lungs into the bloodstream or local tissue. New macromolecule drugs are also being developed for treatment of respiratory diseases such as cystic fibrosis, adult and infant respiratory distress syndromes, asthma and emphysema. This paper reviews formulations for delivering therapeutic aerosols to the lungs. Particular emphasis will be placed on preparing fine powders containing macromolecule drugs. PMID- 10837529 TI - Characterisation of small changes in the physical properties of powders of significance for dry powder inhaler formulations. AB - In this paper we address the following issue: Why is surface characterisation important? All pharmaceutical processes (with the exception of mixing two gasses) involve interfacial contact, and, consequently, it is not surprising that surface energetics play an important role in determining the outcome of all events. For a dry powder inhaler system interfacial considerations may relate to drug-drug interactions (cohesion), drug-carrier or drug-device interactions (adhesion) and deaggregation phenomena during use. As all adhesive and cohesive interactions are interfacial phenomena it is reasonable to accept that the basis of interactions within dry powder inhalers is through interfacial forces, which can be divided into apolar (Lifshitz-van der Waals) and polar (electron donor-electron acceptor) components. Further to this it can be accepted that changes in the nature of any surface within the product (the drug, the carrier or the container) can be expected to result in changes in the surface interactions involving that phase. Thus, in essence, the success or failure of a formulated inhalation device is dependent upon the nature of the surface of the materials used, and, as such, measurement of these surfaces becomes of paramount importance. In this review comparatively little effort will be taken to prove the dominant role of surface energetics in inhalation products; this is primarily because much of the proof which exists is held as confidential by manufacturers. Consequently, this review will concentrate on surface characterisation of powders with respect to determination of surface energies and changes in solid-state properties. PMID- 10837530 TI - Descriptors of irregular particle morphology and powder properties. AB - The description of irregular particle morphology and powder properties may offer unique insights into the performance of materials intended for use in dry powder inhalers. A number of mathematically similar approaches have been used to study these phenomena, all based on evaluating data in terms of irregular oscillations around a mean value. The specific methods described are Fourier, fractal and chaos analysis, stochastic and percolation models. Data describing the morphology of disodium cromoglycate particles, the behaviour of lactose and sodium chloride powders and dispersion of Intal((R)) (disodium cromoglycate/lactose blend) utilizing a Spinhaler((R)) is presented to demonstrate the application of these approaches to dry powder inhaler systems. It is envisaged that these techniques may be used to characterize powders with a view to guiding formulation and that ultimately this may lead to a greater understanding of the nature and magnitude of performance variables. PMID- 10837531 TI - Aerosol particle transport and deaggregation phenomena in the mouth and throat. AB - The results of recent numerical simulations of aerosol particle transport and deaggregation phenomena in the mouth and throat are summarized. These results are used to comment upon the roles played by important factors influencing extrathoracic particle deposition and deaggregation phenomena. The results also provide a basis for comparing the performance of in vitro systems presently used for testing dry powder inhaler devices. Total deposition of aerosol particles in a 'physiologically faithful' three-dimensional (3D) reconstruction of the human mouth and throat closely mimics in vivo deposition data. On the other hand, aerosol particle deposition in a 3D 90 degrees -bend geometry does not accurately simulate aerosol filtration in the human mouth and throat. The numerically predicted regional deposition of aerosol particles closely matches that measured experimentally, in vitro, using the oropharyngeal cast from which the numerical reconstruction was made. A relatively weak dependence of particle deposition on flow-rate is observed with both 90 degrees -bend and 'real' oropharyngeal geometries, inconsistent with extrathoracic deposition in vivo. Two dimensional numerical simulations do not, in general, accurately simulate the conditions in the associated three-dimensional system. Quantitative differences between flow in the 90 degrees -bend and 'real' oropharyngeal geometries are especially significant in regards to the ability of the airstream to break apart particle agglomerates. For both geometries, it is observed that increasingly small particle agglomerates may be separated as the airflow rate increases from 30 to 200 lmin(-1). At flow-rates exceeding 60 lmin(-1), aggregated particles less than approximately 10-um diameter may be separated by the airflow. PMID- 10837532 TI - Review of dry powder inhalers. AB - The search for alternatives to metered-dose inhalers has accelerated recently in a bid to find effective products that do not use chlorofluorocarbon (CFC) propellants. This paper reviews the factors to be considered in developing dry powder inhalers (DPIs), particularly the formulation, metering design and flow path in the device. The advantages and disadvantages of current DPIs are discussed and possible future approaches outlined. PMID- 10837533 TI - A scintigraphic study to evaluate the deposition patterns of a novel anti-asthma drug inhaled from the Cyclohaler dry powder inhaler. AB - A scintigraphic study was performed, in 10 healthy volunteers, to determine the deposition patterns of a novel anti-asthma drug, LAS 31025, inhaled from a dry powder inhaler, the Cyclohaler. In addition, plasma concentrations of drug following inhalation were determined to investigate their relationship to the deposition patterns. Increasing doses of LAS 31025 were inhaled by the volunteers on four separate study days; 2.25, 4.5, 9.0 and 18.0 mg. The average lung deposition values corresponding to these dose levels were 19.1, 20.9, 19.5 and 17.5%, respectively. Deposition patterns within the lungs were similar on each of the four study days, with slightly more of the dose deposited in the central and intermediate regions compared to the peripheral lung region. The scintigraphic data were supported by the pharmacokinetic measurements as there was an increase in the maximum plasma concentration and area under the concentration-time curve corresponding to the increased doses of LAS 31025 on successive study days. PMID- 10837534 TI - Preface. PMID- 10837535 TI - Delivery of molecular and cellular medicine to solid tumors. AB - To reach cancer cells in a tumor, a blood-borne therapeutic molecule or cell must make its way into the blood vessels of the tumor and across the vessel wall into the interstitium, and finally migrate through the interstitium. Unfortunately, tumors often develop in ways that hinder each of these steps. Our research goals are to analyze each of these steps experimentally and theoretically, and then integrate the resulting information in a unified theoretical framework. This paradigm of analysis and synthesis has allowed us to obtain a better understanding of physiological barriers in solid tumors, and to develop novel strategies to exploit and/or to overcome these barriers for improved cancer detection and treatment. PMID- 10837536 TI - Inhibitors of glutathione S-transferases as therapeutic agents. AB - Glutathione S-transferases (GST) are a family of phase II detoxification enzymes with broad substrate specificities. They catalyze the conjugation of glutathione with many different types of xenobiotics, rendering the compound more water soluble and thus more easily eliminated. Resistance to cancer chemotherapeutic drugs, such as the alkylating agents, has been directly correlated with the overexpression of GSTs. Subsequently, a rationale has been established to utilize agents that inhibit GST in combination with alkylating agents to circumvent this resistance. Two such agents, ethacrynic acid (EA) and Terrapin 199 (TER 199), have been examined for this purpose. EA, an inhibitor of all classes of GST isozymes, has been used clinically in combination with thiotepa. More recently, TER 199, a glutathione analog-based GST inhibitor, has been modeled specifically to inhibit GST pi, an enzyme which is commonly found at high levels in human tumor biopsies. Furthermore, a therapeutic strategy has been designed to take advantage of GST pi activation of a prodrug, TER 286. Recent studies have investigated the molecular mechanisms involved in the cellular response to GST inhibitors and have employed techniques such as differential display to examine altered gene expression as well as to identify novel genes induced by these agents. Overall, this strategy may provide further insight into the action of these agents in the cell as well as prove useful in endeavors to modulate anticancer drug resistance. PMID- 10837537 TI - Inhibition of DNA repair as a means of increasing the antitumor activity of DNA reactive agents. AB - Chemotherapeutic alkylnitrosoureas (BCNU, CCNU, streptozotocin) and alkyltriazenes (DTIC, temozolomide) produce a cytotoxic lesion at the O(6) position of guanine. The DNA repair protein, O(6)-alkylguanine-DNA alkyltransferase removes damage from the O(6)-position in a single-step mechanism without co-factors. There is extensive evidence that this protein is one of the most important factors contributing to alkylnitrosourea and alkyltriazene treatment failure. There is an inverse correlation between the level of this protein and the sensitivity of cells to the cytotoxic effects of O(6)-alkylating agents. Attempts have been made to modulate AGT activity using anti-sense technology, methylating agents, O(6)-alkylguanines, and O(6)-benzylguanine analogs. O(6)-Benzylguanine and its analogs are clearly the most potent direct inactivators of the AGT protein. The mechanism involves O(6)-benzylguanine acting as a low-molecular weight substrate with transfer of the benzyl group to the cysteine residue within the active site of the repair protein. Pretreatment of cells with non-toxic doses of O(6)-benzylguanine results in an increase in the sensitivity to O(6)-alkylating agents. Animal studies revealed that the therapeutic index of BCNU increased when administered in combination with O(6) benzylguanine. This drug is currently in phase I clinical trials. Evidence from animal studies indicates that myelosuppression may be the dose-limiting toxicity, thus, efforts are aimed at improving the therapeutic index by the stable expression of O(6)-benzylguanine-resistant AGT proteins into targeted normal tissue such as bone marrow. The successful modulation of alkyltransferases brings on an exciting new era for alkylnitrosoureas and alkyltriazenes. PMID- 10837538 TI - Delivery of tumor suppressor genes to reverse the malignant phenotype. AB - Despite early enthusiasm and excitement, the treatment of cancer via gene therapy is a long way from reaching fruition. The objective of this review is to describe the rationale as to why the delivery of genes encoding functional proteins whose activity has been lost during the initiation and development of cancer may be a feasible therapeutic option. In addition we will evaluate the limitations of the current delivery systems and discuss how these limitations have impacted upon the progress of gene therapy. Finally, we will describe and discuss the most recent attempts to deliver tumor suppressor genes to rodent models of human cancer and perhaps more importantly to human patients. As will become apparent during this review the excitement and enthusiasm for gene therapy remains high, however, this should not diminish the challenges that must be overcome before gene therapy becomes routine. PMID- 10837539 TI - Toward development of a non-viral gene therapeutic. AB - Gene therapy is an emerging field that has reached the early clinical stages of development for some disease states. However, the demonstration of safety in animals and the introduction of gene-based formulations in humans hides the fact that numerous developmental and basic research questions remain. This article highlights progress and emerging issues in the area of liposome-based non-viral gene delivery. The colloidal nature of these formulations render them complicated at the physico-chemical and biological levels. Instrumentation and methodologies need to be developed to better understand the subtleties of plasmid DNA, complexing agents, delivery mode and the route of entry into the cell and the nucleus. Major hurdles to entry include membrane binding, endosomal release, nuclear uptake and decomplexation. Each 'stage' is poorly understood but numerous approaches are being directed to increase cellular delivery. These research efforts, coupled with sensible formulation research and a multi-disciplinary, long-term effort, are necessary for success. PMID- 10837540 TI - Antibody-directed enzyme prodrug therapy (ADEPT): a review. AB - Antibody-directed enzyme prodrug therapy (ADEPT) is a therapeutic strategy which aims to improve the selectivity of anticancer drugs. ADEPT is a two-step antibody targeting system that has benefits over a one-step chemo-, toxin- or radioimmunoconjugate. The basic principles of ADEPT are discussed alongside the requirements of the components: antibodies, enzymes and prodrugs. The design and syntheses of prodrugs are detailed particularly prodrug/drug systems of potential clinical use, the rationale behind their design and the in vitro and in vivo results obtained. The main features of ADEPT, such as targeting of cancer cells by the antibody-enzyme conjugates, enzymic activation of the prodrugs, selection of the prodrug/drug and enzyme/prodrug systems are reviewed. PMID- 10837541 TI - Gene directed enzyme prodrug therapy for cancer. AB - One strategy for gene therapy in malignant disease is gene directed enzyme prodrug therapy (GDEPT). An exogenous enzyme gene is delivered to tumour cells. The enzyme, when expressed, can convert a non-toxic prodrug into a cytotoxic species that is capable of killing the cell in which it has been produced. The most frequently used systems are HSV thymidine kinase with ganciclovir and E. coli cytosine deaminase with 5-fluorocytosine. The bystander effect is of key importance to GDEPT: This describes the local spread of active species from cells that express the enzyme to kill adjacent, untransduced cells. The ultimate success of GDEPT will depend on the ability to achieve efficient gene delivery to and expression in target cells, whilst minimising expression in other tissues. A variety of techniques exist to achieve this goal, including loco-regional administration, manipulation of tumour blood supply and use of tumour-specific promoters to drive enzyme gene expression. PMID- 10837542 TI - Electrochemotherapy: an emerging drug delivery method for the treatment of cancer. AB - The combined treatment consisting of a chemotherapeutic agent and pulsed electric fields has been termed electrochemotherapy. This relatively new treatment modality relies on the physical effects of locally applied electric fields to destabilize cell membranes in the presence of a drug. Membrane destabilization, electroporation, allows increased movement of molecules into the cytosol. Thus, the pulses are used to locally deliver drugs to the interior of cells. This type of treatment has principally been used to deliver bleomycin to tumor cells in vitro and in vivo. Marked antitumor effects have been reported in preclinical studies. In addition, electrochemotherapy clinical trials have been conducted for the treatment of head and neck squamous cell carcinoma, melanoma, and basal cell carcinoma. Objective response rates ranging from 72 to 100% have been reported from these trials. A review of the preclinical and clinical data for this novel drug delivery method is presented. PMID- 10837543 TI - Local delivery of chemotherapy: a supplement to existing cancer treatments. A case for surgical pastes and coated stents. AB - Local application or direct tumor injection of chemotherapeutic drugs has been proposed as a method by which local drug concentrations can be maximized in the immediate tumor environment while systemic exposure and non-target organ toxicity is minimized. Multiple opportunities are available to combine local drug delivery with widely practised, existing medical and surgical therapies. Surgical interventions, including both open and laparoscopic procedures, allow the physician to directly visualize and manipulate pathological tissues. Intraoperative placement of implantable therapeutic compounds (barriers to prevent adhesions, sustained-release antibiotics, tissue 'glues' and hemostatic agents) at or near the disease site is increasingly common in surgical practice. Less invasive therapies assisted by diagnostic imaging (fluoroscopy, ultrasound, CT and MRI scanning) have made accurate needle or catheter placement for drainage (abscesses, cysts, obstructions), injection (contrast media, pharmacological agents, embolic agents) and therapeutic purposes (endoluminal stents, venous filters) widely practised interventional medical procedures. This article describes a chemotherapeutic polymer-based paste we have developed for application at the time of surgery to reduce local recurrence of disease at tumor resection sites and a chemotherapeutic polymer-coated stent for use in the palliative management of malignant obstruction to improve the effective lifespan of the device (e.g., esophageal, biliary, prostate, and pulmonary disease). Despite the growth of local therapy in other disease states, regional cytotoxic drug therapy has not been widely deployed in the management of malignancy due to a clinical bias that local therapy will have limited utility in what is considered to be a systemic disease. In the above manner, local drug delivery could be incorporated into therapeutic protocols designed to enhance, not replace, the efficacy of existing treatment options. PMID- 10837544 TI - Polymeric implants for cancer chemotherapy. AB - Cancer chemotherapy is not always effective. Difficulties in drug delivery to the tumor, drug toxicity to normal tissues, and drug stability in the body contribute to this problem. Polymeric materials provide an alternate means for delivering chemotherapeutic agents. When anticancer drugs are encapsulated in polymers, they can be protected from degradation. Implanted polymeric pellets or injected microspheres localize therapy to specific anatomic sites, providing a continuous sustained release of anticancer drugs while minimizing systemic exposure. In certain cases, polymeric microspheres delivered intravascularly can be targeted to specific organs or tumors. This article reviews the principles of chemotherapy using polymer implants and injectable microspheres, and summarizes recent preclinical and clinical studies of this new technology for treating cancer. PMID- 10837545 TI - Selective boron drug delivery to brain tumors for boron neutron capture therapy. AB - Malignant glioma is one of the most deadly forms of cancer in humans and remains refractory to presently available treatments. Boron neutron capture therapy (BNCT) is a promising therapeutic modality for the treatment of malignant brain tumors. For successful BNCT, a sufficient quantity of boron atoms must be selectively delivered to individual brain tumor cells while at the same time the boron concentration in the normal brain tissue should be kept low to minimize the damage to normal brain tissue. However, the brain entry of drugs is restricted by the blood-brain barrier (BBB), even though the permeability of the pathological area of this barrier may be partially increased due to the present of brain tumors. Therefore, selective delivery of boron to tumor cells across the BBB is a major challenge to the BNCT of brain tumors. This review briefly discusses four main mechanisms responsible for drug transport across the BBB. Brain tumor localizing boron compounds are described, such as borocaptate sodium, p boronophenylalanine, boronated porphyrins and boronated nucleosides. Strategies employed to selectively deliver boron drug into brain tumors are reviewed including hyperosmotic BBB modification, biochemical opening of BBB, electropermeabilization and direct intracerebral delivery of boron drugs. Conjugation of boron drugs to macromolecules like monoclonal antibodies and epidermal growth factor are discussed for active tumor targeting. Boron delivery via microparticles such as liposomes, high density lipoproteins and nanoparticles is also covered for their potential utilization in BNCT of brain tumors. PMID- 10837546 TI - Preface. PMID- 10837547 TI - Self-amplifying vectors for gene delivery. AB - Current methods of gene transfer have a rather low efficiency, especially in vivo. Therefore, one tries to achieve the highest possible levels of expression in the few cells that do take up foreign DNA. One approach is to use self amplifying expression vectors. These vectors are based on the (+)-strand RNA viruses (alphaviruses) Sindbis virus and Semliki Forest virus. In these vectors, the viral capsid protein coding sequences are replaced with the gene of interest. After introduction into the target cells, the viral replication proteins will replicate the recombinant genome. The increased levels of mRNA generate very high transgene expression levels. Furthermore, spread throughout large cells (muscle, neurons) is much better compared to conventional expression cassettes. Self amplifying vectors can be introduced into target cells as RNA, DNA or virions. PMID- 10837548 TI - Cationic lipids used in gene transfer. AB - Gene transfer could represent an important advance in the treatment of both genetic and acquired diseases. Cationic lipid-mediated gene transfer have advantages over viral gene transfer because they are non-immunogenic, they are easy to produce and they are not oncogenic. The major limitation of cationic lipid-mediated gene transfer is its inefficiency. The development of new cationic lipids, and the combination of a cationic lipid with a neutral lipid has conferred improved efficiency to cationic lipid-mediated gene transfer. The mechanism of gene transfer involves entry into the cell via endocytosis, escape from the endocytic compartment, uncomplexing the DNA from the lipids, and transport of DNA to the nucleus. In vivo gene therapy trials for cancer and cystic fibrosis, have shown promising results both in safety and efficacy. PMID- 10837549 TI - Use of the herpes amplicon system as a vehicle for somatic gene transfer. AB - The study of herpes virus biology has produced two vector delivery strategies which exploit the highly evolved neuronal tropism and life cycle regulation exhibited by this class of viruses. Amplicon and recombinant herpes simplex virus systems show promise for use in gene transfer to the nervous system and although neither vector has been used in clinical trials to date, their application to gene delivery and perturbation of neuronal physiology have been well established. We will review the properties of the herpes amplicon system, and demonstrate its utility in neuroscience applications. PMID- 10837550 TI - Progress in development of herpes simplex virus gene vectors for treatment of rheumatoid arthritis. AB - Arthritis is presently incurable and poorly treatable, but there are good grounds for expecting gene therapy to improve matters considerably. Although local ex vivo delivery of anti-arthritic genes to the synovial lining of joints has shown considerable promise, intraarticular gene delivery may be desirable. Herpes simplex virus (HSV) may be a viable vector for in vivo transfer of anti-arthritic genes to joints. HSV has the advantages of high infectivity, large carrying capacity and high titer. The large packaging capacity would permit the inclusion of multiple anti-arthritic genes and necessary regulatory elements. Recombinant vectors produced by this laboratory infect synovial cells efficiently, permitting prolonged expression of transgenes in vitro and in vivo without evidence of cytotoxicity. Further improvements to this vector system include taking advantage of an endogenous HSV 'stealthing' gene, ICP47, which interferes with formation of antigen-class I complexes. Inclusion of inducible promoters to appropriately regulate expression of anti-arthritic genes should further improve this system. PMID- 10837551 TI - Extended release of adenovirus from polymer microspheres: potential use in gene therapy for brain tumors. AB - Current protocols for the treatment of intracerebral glioma are inadequate, with limited reduction in morbidity or mortality. As such, gene therapy paradigms have been developed. Initial progress in rodent models using in situ introduction of retroviral vector producer cells to transfer a cytotoxic gene product led to phase I clinical trials with limited success, due in part to immune responses to murine producer cells along with low level gene transfer. We and others have initiated studies to determine the effectiveness of adenoviral vectors to deliver cytotoxic and/or immuno-stimulatory gene products directly to tumor. Adenoviral vectors can be purified to high titers, are relatively stable upon formulation and storage, and can infect both dividing and non-dividing tumor cells. Also, they can be introduced in situ without helper virus or producer cells. However, gene transfer to glioma tissue with recombinant adenoviruses is not efficient, with multiplicities of infection greater than 50 infectious units/cell required for efficacy. At these doses the virus induces a potent immune response that further reduces gene transfer following re-administration. The inflammatory response to low doses of recombinant adenoviral vectors is less robust and does not preclude re-administration. Thus, strategies to increase efficiency coupled to low dose administration are desirable. To accomplish low dose administration we have developed a method to formulate recombinant adenoviral vectors in biodegradable microspheres. Poly (lactic-glycolic) acid (PLGA) microspheres containing recombinant adenovirus were prepared using a double emulsion technique, and viable virus released for greater than 10 days. PMID- 10837552 TI - Conditionally replicative adenoviruses for cancer therapy. AB - The delineation of the genetic etiology of cancer makes gene therapy a rational approach for the molecular treatment of cancer. Many gene delivery systems have been developed, with viral vectors being the most effective. Underlying cancer gene therapy protocols is the recognition that quantitative tumor transduction cannot be achieved with the vector systems available at the present time. One way to overcome this problem could be to amplify the transduction efficiency through the use of vectors capable of replicating specifically in tumor cells. We are currently developing an adenoviral vector in which viral replication will be restricted to the target tumor cells by limiting the expression of viral genes essential for the virus replication only to the tumor cells of interest. PMID- 10837553 TI - Adeno-associated virus as a gene delivery system. AB - Adeno-associated virus (AAV) has several characteristics which make it extremely attractive as a gene transfer vector: (1) no known pathogenicity; (2) high efficiency and the ability to remain latent; (3) a minimal number of antigens ensuring minimal immunogenicity; (4) the ability to transduce post-mitotic cells; (5) possible advantages of site-specific integration; and (6) a broad host and cell range. The human isolate, AAV-2, is the best studied and has been the focus for gene delivery experiments. This review will discuss recent in vivo experiments demonstrating the utility of AAV in animal models of neurodegenerative disease. PMID- 10837554 TI - Enzyme-catalyzed processes of first-pass hepatic and intestinal drug extraction. AB - Oral bioavailability of pharmacologically effective drugs is often limited by first-pass biotransformation. In humans, both hepatic and intestinal enzymes can catalyze the metabolism of a drug as it transits between the gastrointestinal lumen and systemic blood for the first time. Although a spectrum of drug biotransformations can occur during first-pass, the most common are oxidations catalyzed by cytochromes P450. It is the isozymes CYP2D6, CYP3A4, CYP1A2, CYP2C9 and CYP2C19 that are most often implicated in first-pass drug elimination. For any given substrate, enzyme specificity, enzyme content, substrate binding affinity and sensitivity to irreversible catalytic events all play a role in determining the overall efficiency, or intrinsic clearance, of elimination. Several models have been proposed over the past twenty-five years that mathematically describe the process of drug extraction across the liver. The most widely used, the well-stirred model, has also been considered for depiction of first-pass drug elimination across the intestinal wall. With these models it has been possible to examine sources of interindividual variability in drug bioavailability including, variable constitutive enzyme expression (both genetic and environmentally determined), enzyme induction by drugs, disease and diet, and intrinsic or acquired differences in plasma protein binding and organ blood flow (food and drug effects). In recent years, the most common application of hepatic clearance models has been the determination of maximum organ availability of a drug from in vitro derived estimates of intrinsic metabolic clearance. The relative success of the in vitro-in vivo approach for both low and highly extracted drugs has led to a broader use by the drug industry for a priori predictions as part of the drug selection process. A considerable degree of effort has also been focused on gut wall first-pass metabolism. Important pathways of intestinal Phase II first-pass metabolism include the sulfation of terbutaline and isoproterenol and glucuronidation of morphine and labetalol. It is also clear that some of the substrates for CYP3A4 (e.g., cyclosporine, midazolam, nifedipine, verapamil and saquinavir) undergo significant metabolic extraction by the gut wall. For example, the first-pass extraction of midazolam by the intestinal mucosa appears, on average, to be comparable to extraction by the liver. However, many other CYP3A substrates do not appear susceptible to a gut wall first-pass, possibly because of enzyme saturation during first-pass or a limited intrinsic metabolic clearance. Both direct biochemical and indirect in vivo clearance data suggest significant inter-individual variability in gut wall CYP3A-dependent metabolism. The source of this constitutive variability is largely unknown. Because of their unique anatomical location, enzymes of the gut wall may represent an important and highly sensitive site of metabolically-based interactions for orally administered drugs. Again, interindividual variability may make it impossible to predict the likelihood of an interaction in any given patient. Hopefully, though, newer models for studying human gut wall metabolic extraction will provide the means to predict the average extraction ratio and maximum first-pass availability of a putative substrate, or the range of possible inhibitory or inductive changes for a putative inhibitor/inducer. PMID- 10837555 TI - The effects of diet, aging and disease-states on presystemic elimination and oral drug bioavailability in humans. AB - Presystemic metabolism occurring in the intestinal epithelium and/or liver is frequently an important determinant of drug bioavailability after oral administration. Several factors are potentially involved in such a first-pass effect and their modulation may significantly contribute to intra- and interindividual variability in a drug's plasma concentration-time curve. For example, macronutrient intake and nutritional status may alter cytochrome P-450 (CYP) metabolism by the liver, and food per se in the form of a meal can also affect the first-pass metabolism of some drugs. More important changes, however, result from micronutrients and non-nutrients present in food. In the case of charcoal-broiled and smoked foods, polycyclic aromatic hydrocarbons formed during their preparation result in the induction of xenobiotic metabolizing enzymes, especially those regulated by the Ah-receptor, e.g. CYP1A, which are localized in the intestinal tract. A similar effect also occurs following the ingestion of cruciferous vegetables like brussels sprouts and cabbage, which contain indole related phytochemicals. Such induction can markedly reduce a drug's oral bioavailability. By contrast, the glucosinolate breakdown products of other vegetables produce inhibition of drug metabolism. In the case of phenethyl isothiocyanate-containing watercress, CYP2E1 activity is markedly impaired; however, other organo-sulfur compounds present in, for example, garlic appear to have essentially no effect on drug metabolism. Constituents of grapefruit juice also result in reduced first-pass metabolism, especially for drugs that are CYP3A substrates. Again, this dietary effect is more pronounced in the intestinal epithelium than the liver. A similar, but more generalized, phenomenon also appears to be associated with eating piperine- and capsaicin-containing spices. Possible future applications of such metabolic inhibition include the use of active phytochemicals as bioavailability enhancers for drugs exhibiting a large first-pass effect, and also as cancer chemoprotective agents where CYP-mediated procarcinogen activation is a critical initial step in carcinogenesis. Aging results in a number of physiological changes that potentially can alter drug metabolism and presystemic elimination. By far the most important of these is a reduction in drug metabolizing enzyme activity. Unfortunately, the extent of this effect appears to be unpredictable, both with respect to a specific drug as well as a particular individual. However, the greatest age-related change in oral bioavailability and plasma concentrations is likely to occur with drugs that exhibit a significant first-pass effect (>80%) in young subjects. A similar situation also appears to apply when liver disease is present, especially when this is severe. A further complication in such patients is the presence of vascular shunting, which leads to drug-containing blood by-passing functional enzymes. As a result, plasma levels of drugs that normally exhibit marked first pass metabolism may be many-fold higher in cirrhotic patients compared to those with normal liver function. PMID- 10837556 TI - The barrier function of CYP3A4 and P-glycoprotein in the small bowel. AB - CYP3A4 present in small bowel enterocytes can catalyze substantial metabolism of some orally administered drugs and, thus, exerts a first-pass effect. Recent data indicate that the P-glycoprotein (the MDR 1 gene product) in the enterocyte brush border also limits the bioavailability of many of the same drugs that interact with CYP3A. It has been proposed that P-glycoprotein and CYP3A4 may be functionally linked because (a) the two proteins are co-localized within the digestive tract and within enterocytes, (b) they share many of the same substrates and (c) they are co-inducible in response to at least some xenobiotics. There are several potential mechanisms whereby the functions of P glycoprotein and CYP3A4 could be complimentary. First, Pgp may limit absorption in the proximal small bowel, shifting it to more distal, less catalytically efficient segments that contain lower amounts of CYP3A4. Second, Pgp may function to prolong the duration of absorption. This might increase the duration of exposure of drug to and, hence, the extent of metabolism by enterocyte CYP3A4. Finally, Pgp may preferentially remove from the enterocyte primary drug metabolites that are themselves substrates for CYP3A4. This would limit product inhibition and facilitate primary metabolism catalyzed by CYP3A4. Characterization of the roles of CYP3A4 and Pgp in limiting oral drug availability may be aided by recent success in the development of human intestinal cell lines that stably express both CYP3A4 and Pgp. PMID- 10837557 TI - Impact of P450 genetic polymorphism on the first-pass extraction of cardiovascular and neuroactive drugs. AB - This review highlights the present knowledge on the CYP2D6 (sparteine/debrisoquine) and the CYP2C19 (mephenytoin) polymorphisms. The relevant mutations at genomic level affecting protein expression and function and consequences for first-pass metabolism and effects of cardiovascular and neuroactive drugs are highlighted. In vitro techniques for identification of metabolic steps catalyzed by polymorphic enzymes will be discussed as well as drug-drug interactions related to CYP2D6 and CYP2C19. The importance of the CYP2D6 polymorphism arises from the fact that this enzyme, which is involved in metabolism of more than 50 drugs, is not active in about 8% of a Caucasian population. This group is named poor metabolizers in contrast to the remainder of the population called extensive metabolizers. Depending on the pharmacokinetic and pharmacodynamic properties of the administered drug and its metabolites elevated concentrations of the parent compound can result in an increased risk of toxicity or loss of therapeutic effects in poor metabolizers. On the other hand ultrarapid metabolizers of CYP2D6 might require higher doses than recommended in order to achieve therapeutic drug levels. Moreover, consequences of polymorphic CYP2C19 expression, which is not active in 20% of Orientals and 3% of Caucasians, for drug disposition will be outlined. PMID- 10837558 TI - Contributions of hepatic and intestinal metabolism and P-glycoprotein to cyclosporine and tacrolimus oral drug delivery. AB - The objective of this section is to evaluate the contributions of hepatic metabolism, intestinal metabolism and intestinal p-glycoprotein to the pharmacokinetics of orally administered cyclosporine and tacrolimus. Cyclosporine and tacrolimus are metabolized primarily by cytochrome P450 3A4 (CYP3A4) in the liver and small intestine. There is also evidence that cyclosporine is metabolized to a lesser extent by cytochrome P450 3A5 (CYP3A5). Cyclosporine and tacrolimus are also substrates for p-glycoprotein, which acts as a counter transport pump, actively transporting cyclosporine and tacrolimus back into the intestinal lumen. Traditional teaching of clinical drug metabolism has been that hepatic metabolism is of primary importance, and other sites of metabolism play a relatively minor role. It appears as though intestinal metabolism plays a much greater role in the pharmacokinetics of orally administered drugs than previously thought. Intestinal metabolism may account for as much as 50% of oral cyclosporine metabolism. There are at least two components of intestinal metabolism for cyclosporine and tacrolimus, intestinal CYP3A4/CYP3A5 and intestinal p-glycoprotein activities. The quantity of intestinal enzymes, although highly variable, do not appear to be the key to explaining the variability of oral cyclosporine pharmacokinetics in kidney transplant patients. However, the quantity of intestinal p-glycoprotein accounts for approximately 17% of the variability in oral cyclosporine pharmacokinetics. It may be that p glycoprotein maximizes drug exposure to intestinal enzymes, thus decreasing the importance of enzyme quantity. Since cyclosporine's FDA approval in 1983, there have been many reports of clinically significant drug interactions of other agents when given concomitantly with cyclosporine. With the FDA approval of tacrolimus in 1994, a similar pattern of clinically significant drug interactions appears to be emerging. It seems that compounds that alter (either induce or inhibit) CYP3A4 and/or p-glycoprotein will alter the oral pharmacokinetics of cyclosporine and tacrolimus. It should be expected that, until further data are available, the drugs which interact with cyclosporine will also interact with tacrolimus. PMID- 10837559 TI - Human immunodeficiency virus protease inhibitors. From drug design to clinical studies. AB - The discovery of human immunodeficiency virus (HIV) protease inhibitors is an example in which pharmacokinetic evaluation was implemented early in the discovery phase to obtain optimal pharmacological and pharmacokinetic properties. Currently, three HIV protease inhibitors, saquinavir, indinavir and ritonavir are clinically available. As a family, these HIV protease inhibitors are characterized pharmacologically by their ability to inhibit the viral protease enzyme. Pharmacokinetically, they are quite different due to their dissimilarity in physicochemical properties. Bioavailability appears to be limited with saquinavir, but not with indinavir and ritonavir. Although all three drugs are metabolized extensively by cytochrome P-450, saquinavir and indinavir are high clearance drugs while ritonavir is a low clearance drug. Despite their significant differences in elimination clearance, all three HIV proteases are given at high oral doses (600-800 mg) either twice or three times daily. These HIV protease inhibitors show superior therapeutic activity and a more favorable safety profile than those reported for the established reverse transcriptase inhibitors. However, the potential for interactions with other drugs metabolized by the CYP 3A4 isoform appears to be considerable. In addition, repeated administration of enzyme inducers results in a substantial decrease of plasma concentrations of protease inhibitors. Therefore, co-administration of drugs, such as rifampicin and rifabutin, must be avoided. HIV protease inhibitors are promising in the treatment of AIDS. Although they are not a cure, they can significantly inhibit that viral replication and improve the quality of life for people who have HIV infection. PMID- 10837560 TI - Improvement of oral peptide bioavailability: Peptidomimetics and prodrug strategies. AB - Clinical development of orally active peptide drugs has been restricted by their unfavorable physicochemical properties, which limit their intestinal mucosal permeation and their lack of stability against enzymatic degradation. Successful oral delivery of peptides will depend, therefore, on strategies designed to alter the physicochemical characteristics of these potential drugs, without changing their biological activity, in order to overcome the physical and biochemical barrier properties of the intestinal cells. This manuscript will focus on the physiological limitations for oral peptide delivery and on various strategies using chemical modifications to improve oral bioavailability of peptide-based drugs. PMID- 10837561 TI - Preface. PMID- 10837562 TI - Biodegradation and biocompatibility of PLA and PLGA microspheres. AB - A fundamental understanding of the in vivo biodegradation phenomenon as well as an appreciation of cellular and tissue responses which determine the biocompatibility of biodegradable PLA and PLGA microspheres are important components in the design and development of biodegradable microspheres containing bioactive agents for therapeutic application. This chapter is a critical review of biodegradation, biocompatibility and tissue/material interactions, and selected examples of PLA and PLGA microsphere controlled release systems. Emphasis is placed on polymer and microsphere characteristics which modulate the degradation behaviour and the foreign body reaction to the microspheres. Selected examples presented in the chapter include microspheres incorporating bone morphogenetic protein (BMP) and leuprorelin acetate as well as applications or interactions with the eye, central nervous system, and lymphoid tissue and their relevance to vaccine development. A subsection on nanoparticles and nanospheres is also included. The chapter emphasizes biodegradation and biocompatibility; bioactive agent release characteristics of various systems have not been included except where significant biodegradation and biocompatibility information have been provided. PMID- 10837563 TI - Preparation of microspheres by the solvent evaporation technique. AB - The microencapsulation process in which the removal of the hydrophobic polymer solvent is achieved by evaporation has been widely reported in recent years for the preparation of microspheres and microcapsules based on biodegradable polymers and copolymers of hydroxy acids. The properties of biodegradable microspheres of poly(lactic acid) (PLA) and poly(lactic-co-glycolic acid) (PLGA) have been extensively investigated. The encapsulation of highly water soluble compounds including proteins and peptides presents formidable challenges to the researcher. The successful encapsulation of such entities requires high drug loading in the microspheres, prevention of protein degradation by the encapsulation method, and predictable release of the drug compound from the microspheres. To achieve these goals, multiple emulsion techniques and other innovative modifications have been made to the conventional solvent evaporation process. PMID- 10837564 TI - One- and three-month release injectable microspheres of the LH-RH superagonist leuprorelin acetate. AB - The biodegradable polymers poly(lactic/glycolic acid) (PLGA) and poly(lactic acid) (PLA) were used as wall materials in the preparation of microspheres (msp) containing the LH-RH superagonist leuprorelin (leuprolide) acetate. A novel W/O/W emulsion-solvent evaporation method was devised for the preparation of msp containing this water-soluble peptide. This method achieved high entrapment efficiency and sustained drug release over a long period predominantly due to polymer bioerosion. The msp are fine microcapsules with polycores containing the peptide at a high concentration and are easily injectable through a conventional fine needle. Leuprorelin msp made with PLGA(75/25)-14,000 or PLA-15,000 released the drug in a zero-order fashion, maintained constant serum drug levels and attained persistent objective suppression of the pituitary-gonadal system ('chemical castration') over 1 or 3 months after i.m. or s.c. injection into animals. These results indicate that depot formulations may be potentially useful in the therapy of endocrine diseases in humans. In this paper, studies on the formulation, drug release and pharmacological effects in animals for these leuprorelin depot formulations are reviewed. PMID- 10837565 TI - Recombinant human growth hormone poly(lactic-co-glycolic acid) microsphere formulation development. AB - The development of a sustained release formulation of recombinant human growth hormone (rhGH) has focused on a depot preparation using the biodegradable polymer, poly(lactic-co-glycolic acid) (PLGA), for microsphere production. These formulations have been designed to assure the maintenance of protein integrity both during the microencapsulation process and upon subsequent release in vitro and in vivo. In addition, animal models were developed to assess both the in vivo release kinetics and the potency of the released protein. These studies emphasized the importance of obtaining a correlation between the in vivo and in vitro release at an early stage of development. Juvenile rhesus monkey studies revealed that continuous rhGH administration resulted in a greater total insulin like growth factor-I (IGF-I) response than daily rhGH administration, indicating that a continuous rhGH dose may provide comparable efficacy to daily dosing at a lower total dose of rhGH. The use of a conventional water-in-oil-in-water process yielded a triphasic release of biologically active and non-immunogenic rhGH, while the novel cryogenic process achieved a continuous release of rhGH that is biologically active and non-immunogenic. The rhGH PLGA formulation produced by the novel cryogenic process was manufactured under aseptic GMP conditions and was shown to be safe in growth hormone-deficient adults. This protein and these studies should serve as a model for the future development of PLGA formulations for therapeutic proteins. PMID- 10837566 TI - Multiple emulsion technology for the design of microspheres containing peptides and oligopeptides. AB - This paper reviews the preparation and characterization of small poly(lactic-co glycolic acid) microspheres (mean size lower than 10 um) containing small peptides and prepared by a water-in-oil-in-water emulsion solvent evaporation technique. These microspheres were shown to encapsulate very efficiently a 33 amino acid peptide (V3 BRU) and in vitro release kinetics studies showed that such microspheres could be employed for both oral vaccination and controlled release. The encapsulation of a seven amino acid peptide (pBC 264) led on the contrary to a very low encapsulation efficiency. In order to increase the encapsulation of pBC 264, two strategies were adopted: (i) taking into account the solubility of pBC 264 at different pH, the inner aqueous phase was maintained at a basic pH where the peptide was soluble, while the external aqueous phase was acidic; (ii) ovalbumin was added to stabilize the inner emulsion. These two strategies allowed us to increase significantly the encapsulation rate of pBC 264. Nevertheless, the in vitro release kinetics of the peptide were strongly influenced by the presence of ovalbumin which seems to form pores in the microsphere structure. By contrast, when ovalbumin was replaced by Pluronic(R) F68, microspheres did not have pores, thus the release profile and the extent of the burst were much smaller. When microspheres were stereotactically implanted in the rat brain, in vivo release profiles were in good agreement with the release observed in vitro. In conclusion, these microspheres are well suited for the slow delivery of neuropeptides in the brain, a feature expected to facilitate the study of long term effects of these compounds. PMID- 10837567 TI - New advances in microsphere-based single-dose vaccines. AB - Polymer microspheres have shown great potential as a next generation adjuvant to replace or complement existing aluminum salts for vaccine potentiation. Microsphere-based systems can now be made to deliver subunit protein and peptide antigens in their native form in a continuous or pulsatile fashion for periods of weeks to months with reliable and reproducible kinetics, often obviating the need for booster immunizations in animal models. Microspheres have also shown potential as carriers for oral vaccine delivery due to their protective effects on encapsulated antigens and their ability to be taken up by the Peyer's patches in the intestine. The potency of these optimal depot formulations for antigen may be enhanced by the co-delivery of vaccine adjuvants, including cytokines, that are either entrapped in the polymer matrix or, alternatively, incorporated into the backbone of the polymer itself and released concomitantly with antigen as the polymer degrades. In this article we review the use of polymer microspheres for single-step immunization and discuss future applications for the improvement of vaccines and immunotherapies by utilizing encapsulation technology. PMID- 10837568 TI - Therapeutic effects of leuprorelin microspheres in prostate cancer. AB - Leuprorelin has demonstrated effectiveness comparable to orchiectomy and oral diethylstilboestrol for the palliation of advanced prostate cancer. Unlike orchiectomy, leuprorelin's effects are reversible; also leuprorelin is not associated with the cardiovascular or thromboembolic adverse effects of oestrogens. For these reasons, leuprorelin has been widely used as an alternative to surgical castration or to oestrogens in the treatment of metastatic prostate cancer. Sustained-release leuprorelin microsphere formulations have been developed which exhibit zero order release of active drug from the injection site, such that in the United States the 7.5 mg dosage strength is recommended to be administered once a month and the 22.5 mg dosage strength once every three months. Although most patients will have suppressed release of pituitary luteinizing hormone by the third or fourth week after the first dose of depot leuprorelin, 4-5% of treated patients have been reported to have delayed responses, taking many more weeks or months to respond. A transient biochemical hormone escape has also been reported, although worsening of clinical symptoms has not accompanied the elevation of serum testosterone levels during treatment. Usually, leuprorelin is initiated as monotherapy when patients with advanced prostate cancer become symptomatic. However, newer studies of combination therapy of luteinizing hormone releasing hormone analogs with antiandrogens suggest that early initiation of therapy, at the time of diagnosis of advanced disease, may be beneficial, particularly in a subgroup of patients with small volume disease and good performance status. Leuprorelin is also undergoing evaluation as neoadjuvant therapy prior to radical prostatectomy for localized prostate cancer. Preliminary studies suggest that neoadjuvant leuprorelin in combination with an antiandrogen may be effective in downstaging prostate tumours. Leuprorelin commonly produces several adverse effects: hot flashes, decreased libido and impotence, and tumour flare. PMID- 10837569 TI - Therapeutic effects of leuprorelin microspheres on endometriosis and uterine leiomyomata. AB - Uterine leiomyomata and endometriosis are two common hormone-dependent pathologies effecting women of reproductive age. Prior to the introduction of gonadotropin releasing hormone agonists (GnRHa), there was no effective medical therapy for leiomyomata, and the most effective pharmaceutical intervention for endometriosis had unacceptable side effects. Early intranasal and non-depot formulations of GnRHa were effective treatments for these diseases but patient acceptance and compliance was poor. The introduction of depot microspheres of leuprorelin (leuprolide) acetate has provided efficacious and safe medical management of both endometriosis and uterine leiomyomata which is well tolerated by most patients. PMID- 10837570 TI - Rational use of agonists and antagonists of luteinizing hormone-releasing hormone (LH-RH) in the treatment of hormone-sensitive neoplasms and gynaecologic conditions. AB - Analogues of luteinizing hormone-releasing hormone (LH-RH) have made possible new approaches to the treatment of some hormone-dependent cancers and diseases and conditions which result from inappropriate sex hormone levels. In the fields of both gynaecology and oncology, the development of sustained delivery depot systems has played a key role in the clinical use of LH-RH agonists and will be also essential for the LH-RH antagonists. Clinical results show that therapy with agonists of LH-RH is the preferred method of treatment for men with advanced prostate cancer. For prostate cancer and other indications, the new LH-RH antagonists such as Cetrorelix may offer an advantage based on the fact that they inhibit LH, FSH and sex-steroid secretion from the start of the administration and thus reduce the time of the onset of therapeutic effects. The use of antagonists would avoid the temporary clinical "flare-up" of the disease which can occur with the agonists in men with prostate cancer. The rapid shrinkage of the prostate and improvement in urinary symptoms obtained with Cetrorelix in men with benign prostatic hyperplasia (BHP) suggests that LH-RH antagonists offer a therapeutic alternative in patients who are considered poor surgical risks. Various experimental and clinical studies suggest that analogues of LH-RH might be useful for treatment of premenopausal women with oestrogen-dependent breast cancer. LH-RH antagonists such as Cetrorelix could be also considered for hormonal therapy of epithelial ovarian cancer which responds only marginally to the agonists, and for treatment of endometrial cancer. Many investigators have reported beneficial effects of LH-RH agonists in the treatment of patients with leiomyomas. LH-RH antagonists also appear to be promising for therapy of uterine leiomyomas, and in addition might be useful for treatment of endometriosis and polycystic ovarian disease (PCOD). LH-RH agonists have been employed in in vitro fertilization and embryo transfer (IVF-ET) programs to prevent a premature rise in LH and various results suggest that the use of antagonist Cetrorelix in assisted reproduction procedures, could be even more advantageous. For most of these indications, the use of sustained release depot preparations will be required. PMID- 10837571 TI - Preface. PMID- 10837572 TI - Intraruminal devices. AB - Intraruminal boluses are drug delivery devices designed to provide long term delivery of drug to the reticulo-rumen compartments of ruminant animals. This cavity is the site of the bacterial fermentation and breakdown of foodstuffs and represents an ideal site for the long term delivery of drugs and nutrients to these grazing animals. Often such animals are 'turned out' in the spring to spend much of the spring, summer and autumn seasons grazing open range. As such, the administration of drugs during the grazing season is complicated by the need to first retrieve the animals prior to drug administration. Long term delivery of drug circumvents this problem providing for decreased costs and improved efficiencies for the farmer. Numerous types of delivery devices have been developed to meet this need. This review summarizes the various approaches that have been developed, provides information on the critical design features and reviews the relevant anatomy and physiology of these animals. Some comments are provided regarding needed future developments for this class of drug delivery system. PMID- 10837573 TI - Rumen-stable delivery systems. AB - Ruminants have a distinct digestive system which serves a unique symbiotic relationship between the host animal and predominantly anaerobic rumen bacteria and protozoa. Rumen fermentation can be both beneficial by enabling utilization of cellulose and non-protein nitrogen and detrimental by reducing the nutritive value of some carbohydrates, high biological value proteins and by hydrogenating unsaturated lipids. In addition it can also result in the modification and inactivation of many pharmacologically active ingredients administered to the host animal via the oral route. The advances in ruminant nutrition and health demand a rumen-stable delivery system which can deliver the active ingredient post-ruminally while simultaneously meet efficacy, safety and cost criteria. In contrast to drug delivery systems for humans, the demand for low-cost has hindered the development of effective rumen-stable delivery systems. Historically, heat and chemical treatment of feed components, low solubility analogues or lipid-based formulations have been used to achieve some degree of rumen-stability, and products have been developed accordingly. Recently, a polymeric pH-dependent rumen-stable delivery system has been developed and commercialized. The rationale of this delivery system is based on the pH difference between ruminal and abomasal fluids. The delivery system is composed of a basic polymer, a hydrophobic substance and a pigment material. It can be applied as a coating to solid particles via a common encapsulation method such as air-suspension coating. In the future, the delivery system could be used to deliver micronutrients and pharmaceuticals post-ruminally to ruminant animals. A further possible application of the delivery system is that it could also be combined with other controlled delivery devices/systems in order to enhance slow release or to achieve targeted delivery needs for ruminants. This paper discusses the rumen protection and the abomasal release mechanism of the polymeric coating. It also reviews other rumen stable delivery systems and methods for evaluating their in vitro and in vivo performance. PMID- 10837574 TI - Ocular drug delivery in veterinary medicine. AB - This paper provides a comprehensive overview of the various approaches currently used in the development of ocular drug delivery systems for the treatment of ocular diseases in animals. It is obvious from the literature that most of the products that are currently available are derived from human medicine without consideration given to the differences which exist between the anatomy and physiology of the eye of various animal species which ultimately affect product design and performance. As a result, many of the products for animal use seem in many circumstances inappropriate for animal care. The article deals with some aspects of eye anatomy and physiology of different animals, and then provides an overview of the most commonly encountered pathologies. The paper then discusses the currently available drug products and finally reviews new delivery concepts. Several hundred references are included in the paper and provide access to further information on the subject. PMID- 10837576 TI - Preface. PMID- 10837575 TI - Conceptual and commercially available intravaginal veterinary drug delivery systems. AB - In the veterinary area the utilization of the vagina as a route for drug delivery has focused on the systemic delivery of hormonal steroids to control synchrony, ovulation and fertility in a variety of livestock. There are several reasons for administering drugs to modify the menstrual cycle in humans and the estrous cycle in animals and this paper provides a comparison of the objectives of drug intervention to control reproduction in humans and livestock. In addition, the types of hormones and their concentrations found during the menstrual cycle in humans and during the estrous cycle in animals are compared. The paper also reviews the intravaginal drug delivery systems developed for the control of the estrous cycle in livestock and both conceptual and commercially available intravaginal drug delivery systems that have been described in the literature are described. The history of some of the delivery systems is included and the future directions of this area of research are discussed. PMID- 10837577 TI - Anatomy, physiology and function of the nasal cavities in health and disease. AB - In studying drug absorption from the nasal mucous membrane, it is essential to have a clear understanding of anatomy and physiology of the nose, and how it relates to the characteristics of the delivery system used. The human nose is characterized by an individually varying shape and caliber, which might interfere with standard recommendations of intranasal medication. It is also of significance that there is a tendency for reflex- induced and profuse watery hypersecretion from glands, and for quick and considerable changes of mucosal thickness due to the presence of large venous sinusoids. These are factors which can interfere with pharmacokinetics. Also mucociliary transport rate has to be taken into consideration, as the drug is removed from the absorptive mucous membrane within 30 min. Intranasal drug distribution has been poorly studied in relation to nasal anatomy and pathology. In contrast to common believe, nasal inflammation does not seem to increase drug absorption. On the contrary, blockage, sneezing and rhinorrhoea might preclude the absorption. PMID- 10837578 TI - Nasal mucociliary clearance as a factor in nasal drug delivery. AB - The nasal mucociliary clearance system transports the mucus layer that covers the nasal epithelium towards the nasopharynx by ciliary beating. Its function is to protect the respiratory system from damage by inhaled substances. Impairment of nasal mucociliary clearance can result in diseases of the upper airways. Therefore, it is important to study the effects of drugs and drug excipients on nasal mucociliary clearance. A large number of methods are used to assess mucociliary clearance. These methods study the effects of drug and excipients on the mucociliary system in vitro or in vivo in animals and humans. In some cases, the results of different in vitro and in vivo measurements do not correlate well. In vitro methods, especially ciliary beat frequency measurements, have been demonstrated to be valuable tools for toxicity screening. However, in vivo studies are essential to confirm the safety of nasal drug formulations. Nasal mucociliary clearance also has implications for nasal drug absorption. Drugs are cleared rapidly from the nasal cavity after intranasal administration, resulting in fast systemic drug absorption. Several approaches are discussed to increase the residence time of drug formulations in the nasal cavity, resulting in improved nasal drug absorption. However, more experimental evidence is needed to support the conclusion that this improved absorption is caused by a longer residence time of the nasal drug formulation. PMID- 10837579 TI - Intranasal drug delivery. AB - In recent years the nasal route has received a great deal of attention as a convenient and reliable method for the systemic administration of drugs. Although this route is currently being marketed for the systemic administration of several drugs, it has only recently been studied. This chapter deals with animal techniques to study nasal absorption and the effect of physico-chemical and biopharmaceutical properties of drugs on the rate and extent of absorption. It also discusses factors affecting peptide absorption and methods to improve the nasal bioavailability of peptides. In the last part of the chapter the utility of the nasal route for the delivery of drugs to the brain is discussed. PMID- 10837580 TI - In vitro cell models to study nasal mucosal permeability and metabolism. AB - Whereas in vivo studies represent the most crucial test for any nasal drug application or formulation, mechanistic aspects of nasal absorption may be more clearly approached by well defined and controlled in vitro studies. In this review the progress of nasal in vitro models to investigate drug permeation and metabolism in the epithelium is summarized and their potential and limitations are discussed. The following subjects will be covered: (i) primary cell cultures of human nasal epithelium, including sampling techniques and culture conditions, (ii) human nasal cell lines (in particular the human nasal cell line RPMI 2650), and (iii) excised nasal epithelium (rabbit, bovine, ovine, canine, human), also summarizing suitable preparation techniques and tissue characterization, test media, tissue equilibration, viability testing, and integrity tests. Furthermore, an overview on the various experimental set-ups suitable for in vitro transport studies (permeation rates; identification of permeation pathways; mechanisms and toxicity of absorption enhancers) and for metabolism studies (rates, saturation and pathways of enzymatic cleavage) is presented. Some attention is given to identify potential endocytotic uptake mechanisms. To date, the permeation and metabolic barrier function of excised nasal tissue derived from various animals has shown to mimic the in vivo situation 'ex vivo' at the highest degree possible. Supply of human tissue will continue to be short. Therefore, further studies are necessary to evaluate and improve culture conditions, handling, performance and physiologic relevance of primary human cell and cell line cultures. PMID- 10837581 TI - Peptide hormones: Review of current and emerging uses by nasal delivery. AB - The family of clinically available peptide hormones (PHs) is expanding in an exponential way, and advancement of knowledge of the basic mechanisms of action of PHs has led to multiplication of the possible clinical indications of already known PHs, and appears even more promising for still unknown PHs. A common obstacle to a full routine use of PHs is represented by the fact that PHs cannot be administered by the oral route, since they undergo digestion and inactivation in the gastrointestinal tract and a significant first pass metabolism in the liver. One alternative is represented by intranasal administration of PHs. The intranasal route of administration of PHs is also very attractive because of its convenience, which should assure a good compliance by patients. Luteinizing hormone releasing hormone, the analogues, desmopressin, oxytocin and salmon calcitonin are already marketed for intranasal administration; for salmon calcitonin, studies about bioavailability have been scanty in the past, but should be re-considered in order to fully explore its clinical benefit.Intranasal peptide hormones not yet on the market are insulin, glucagon, growth hormone releasing hormone (GHRH) and GHRP, GH and somatostatin, but the scenario is likely to change in a short period of time. Hexarelin seems very effective and is at a promising stage of development; also, glucagon appears mature enough to undergo extensive clinical evaluation and possibly marketing. The concern is why other peptides have not been further evaluated, as is the case for somatostatin and its analogues. PMID- 10837582 TI - Effects of physicochemical properties and other factors on systemic nasal drug delivery. PMID- 10837583 TI - Optimization of systemic nasal drug delivery with pharmaceutical excipients. PMID- 10837584 TI - Prodrugs for nasal drug delivery. AB - Recently, the delivery of xenobiotics via the nasal route has received increasing attention as this offers several advantages, i.e. high systemic availability, rapid onset of action. Both charged and uncharged forms of drugs can be transported across the nasal epithelium. This mucosa is rich in various metabolizing enzymes such as aldehyde dehydrogenase, glutathione transferases, epoxide hydrolases, cyt-P450-dependent monooxygenases. The presence of these enzymes may make it possible for pharmaceutical scientists to design prodrugs for better absorption and high systemic availability. Recent advances in peptide nasal delivery through prodrug modification has been thoroughly discussed in this paper. Finally, nasally delivered therapeutic agents targeted to various disease states have been examined. PMID- 10837585 TI - Large molecule and particulate uptake in the nasal cavity: the effect of size on nasal absorption. AB - One of the characteristics influencing the increased interest in the nasal cavity as a site for systemic drug delivery is the ability of large molecules to permeate through the nasal mucosa into the systemic circulation. Compilations of data regarding the absorption of large therapeutic agents, peptides and proteins in particular, along with more systematic studies using polymeric compounds have shown that for compounds larger than 1000 Da, bioavailability can be directly predicted from a knowledge of molecular weight. In general, the bioavailability of these large molecules ranges from 0.5 to 5%. Particulate uptake also occurs in the nasal mucosa, and particles up to approximately 1 um have been shown to rapidly enter the bloodstream following intranasal administration. The unique barrier properties of this mucosal delivery site give it great promise as a route for the systemic administration of large molecules. PMID- 10837586 TI - Nasal delivery systems and their effect on deposition and absorption. AB - Due to nasal anatomy and physiology, with a non-ciliated area in the anterior part of the nasal cavity and a ciliated region in the more posterior part of the nose, the site of deposition is of importance for the nasal mucociliary clearance and retainment of a formulation in the nose. Many drug delivery devices for nasal application of liquid, semisolid and solid formulations were investigated in respect to their deposition in the nasal cavity. The site of deposition and the deposition area depend on several parameters which are related to the delivery device, such as mode of administration, particle size of the formulation and velocity of the delivered particles. Several in vitro and in vivo methods have been used to study distribution and clearance of intranasally delivered therapeutics. The relationship between deposition, absorption and related bioavailability of the nasally applied formulation has been shown. PMID- 10837587 TI - Mathematical modeling of deposition and disposition of drugs administered via the nose. AB - This articles reviews the mathematical models of deposition and disposition of drugs administered into the human nasal cavity for systemic activity. The modeling of the disposition kinetics includes drug release from carriers, translocation within the nasal cavity and into the gastrointestinal tract, drug decomposition, and drug absorption during the transit through the nose and the gastrointestinal tract. The uses of such mathematical models for design and analysis of nasal delivery systems are illustrated, with particular reference to the new therapeutic materials and excipients. PMID- 10837588 TI - Microspheres as nasal drug delivery systems. AB - All types of microspheres that have been used as nasal drug delivery systems are water-insoluble but absorb water into the sphere's matrix, resulting in swelling of the spheres and the formation of a gel. The building materials in the microspheres have been starch, dextran, albumin and hyaluronic acid, and the bioavailability of several peptides and proteins has been improved in different animal models. Also, some low-molecular weight drugs have been successfully delivered in microsphere preparations. The residence time in the cavity is considerably increased for microspheres compared to solutions. However, this is not the only factor to increase the absorption of large hydrophilic drugs. Microspheres also exert a direct effect on the mucosa, resulting in the opening of tight junctions between the epithelial cells. Starch and dextran microspheres have been administered repeatedly and can be classified as safe dosage forms. PMID- 10837589 TI - Preface. PMID- 10837590 TI - The biochemical and physiological characteristics of receptors. AB - Cell surface receptors play a central role in the regulation of both cellular and systemic physiology by mediating intercellular communication, facilitating protein trafficking, and regulating virtually all intracellular processes. Receptor expression is often cell specific and is determined by cellular lineage, genetics, and a variety of factors in the extracellular milieu. As receptors are generally localized on the plasma membrane and differentially expressed in certain cell types and tissues, they provide a potential target for drug delivery. However, since most receptors are integrally connected with intracellular signal transduction networks, targeting via these receptors may elicit a biological response. This review describes some established and emerging concepts regarding the structure and functions of receptors. In addition, some aspects related to the regulation and crosstalk between receptors are discussed. PMID- 10837591 TI - Endocytosis and transcytosis. AB - Vesicular coat proteins mediate the formation of nascent vesicles and select the cargo to be incorporated therein. As additional coat proteins are discovered that regulate vesicular traffic along very specific intracellular pathways, the possibility looms of regulating the intracellular trafficking and targeting of therapeutic agents by modulation of the action of vesicular coat proteins. Examples are provided of coat proteins thought to regulate the trafficking of pharmaceutically relevant molecules via clathrin-mediated endocytosis, caveolae mediated endocytosis, and transcytosis. PMID- 10837592 TI - Molecular motors and their role in membrane traffic. AB - Recent investigations support a role for the vesicle motor proteins (kinesin, cytoplasmic dynein, and myosin) in numerous membrane trafficking events including endocytosis and transcytosis. Kinesin and cytoplasmic dynein are responsible for movement of membrane vesicles along cellular microtubules to and from cellular membrane compartments, while certain members of the myosin family also appear to drive membrane vesicles along actin filaments to and from membrane compartments. In this review, our current understanding of the role of these vesicle motors in membrane trafficking is highlighted. Future areas of interest which may be able to make use of these vesicle motors as potential targets for drug delivery are also discussed. PMID- 10837593 TI - Receptor-mediated delivery of foreign genes to hepatocytes. AB - Naturally existing cell surface receptors provide convenient systems for transporting substances from the extracellular environment to the interior of cells. When such receptors are highly selective for certain cell types, the possibility exists for targeting biological agents of interest to specific tissues. In this review, the concepts, progress and problems in targeting genes and their subsequent expression in hepatocytes based on the asialoglycoprotein receptor in vitro and in vivo will be discussed. The use of protein carriers for hepatocyte specific delivery of double stranded DNA to introduce novel gene expression novel, as well as, delivery of single stranded DNA for inhibition of endogenous genes with examples of each are reviewed. PMID- 10837594 TI - Ligand-targeted liposomes. AB - Liposomes have gained increased attention as systemic drug delivery vehicles following recent regulatory approvals of several vesicle-formulated drugs. These products have demonstrated improved therapeutic indices over their corresponding conventional drugs by avoiding sensitive tissues and/or increasing delivery to specific targets in vivo. They have achieved these improvements primarily through physical means: (1) by retaining drug within vesicles while in the circulation, thus avoiding or minimizing uptake by sensitive normal tissues; and (2) by selectively extravasating into target tissues, releasing active drug. In order to improve upon these therapies in the future, clinically active liposome delivery systems most likely will need to include site-directed surface ligands to further enhance their selective delivery. This may be crucial for the in vivo transport and delivery of macromolecules, including antisense, oligonucleotide aptamers, and genes, which-unlike most conventional drugs-do not circulate well and often require cellular uptake by fusion, endocytosis, or other processes to reach their active sites. This manuscript reviews technologies applicable to directing liposomes and their contents to selected in vivo targets using surface-bound, site-specific ligands. Presented are the biological barriers to be overcome, criteria for selecting the determinants to be targeted, various targeting ligands and overall delivery system design considerations. Several novel targets as well as novel ligand constructs for site-directed therapy are reviewed and discussed. Systemic liposome therapy, which currently must be administered by the intravenous route, is the principal focus of this analysis. PMID- 10837595 TI - Receptor-mediated targeted drug or toxin delivery. AB - The new approach to the treatment of cancer or to immunomodulation is drug targeting. Cellular uptake of drugs bound to a targeting carrier or to a targetable polymeric carrier is mostly restricted to receptor-mediated endocytosis. Factors that influence the efficiency of receptor-mediated uptake of targeted drug conjugate are the affinity of the targeting moieties, the affinity and nature of the target antigen, density of the target antigen, the epitope of the target antigen, the type of cell target, the rate of endocytosis, the route of internalization of the ligand-receptor complex, the ability of the drug or toxin to release from its targeted carrier, the ability of the drug or toxin to escape from a vesicular compartment into the cytosol, the affinity of the carrier to the drug and the concentration of the carrier. Targeted chemotherapy is also significantly influenced by the antigenic modulation and/or immunoselection of tumor cells. The binding of drug (toxin) to targetable polymeric carrier considerably decreases unwanted side toxicity. PMID- 10837596 TI - Preface. PMID- 10837597 TI - Gene delivery systems: Bridging the gap between recombinant viruses and artificial vectors. AB - Although most research in the field of somatic gene therapy has investigated the use of recombinant viruses for transferring genes into somatic target cells, various methods for nonviral gene delivery have also been proposed. Both types of gene delivery systems have advantages and drawbacks. Schematically, viral vectors are particularly efficient for gene delivery, whereas nonviral systems are free of the difficulties associated with the use of recombinant viruses but need to be further optimized to reach their full potential. In order to bridge the gap between viral vectors and synthetic reagents, we discuss here some specific features of the viral vector systems of today that could advantageously be taken into account for the design of improved nonviral gene delivery systems. Indeed, although nonviral systems differ fundamentally from viral systems, one possible approach towards enhanced artificial reagents aims at developing 'artificial viruses' that mimic the highly efficient processes of viral infection. PMID- 10837598 TI - Assaying extrachromosomal gene therapy vectors that carry replication/persistence elements. AB - Persistence in the cell is a desirable property for most gene therapy vectors. For extrachromosomal vectors, persistence is limited in most cell types. To address this problem, we have developed vectors with the ability to replicate and be retained in the nucleus. These properties are conferred by specific elements present on the vectors and derived from genomic DNA and from Epstein-Barr virus. In order to begin evaluation of these vectors for use in gene therapy, we developed and present here two assays that measure the persistence of vector DNA in tissue culture cells under rapidly dividing and slowly dividing conditions. Our results indicate that inclusion of DNA replication and nuclear retention elements on a vector increases persistence of vector DNA in slowly dividing cells by at least 500%. Further improvement of the system is discussed. PMID- 10837599 TI - A novel RU486 inducible system for the activation and repression of genes. AB - We have developed an inducible system that consists of a transactivator and a target gene. The transactivator encodes a chimeric regulator that is responsive to RU486 (mifepristone, a progesterone receptor antagonist) but not to progestins and other hormones or endogenous ligands for activation. The target gene can be any gene under the control of Gal4 DNA binding sites. When the regulator is activated by RU486, it induces target gene expression by binding to the Gal4 recognition sequences upstream of the target. To verify this concept, we have successfully demonstrated the functionality of this system in tissue culture and in transgenic mice. Furthermore, for applications that require higher levels of a target gene, we also have generated regulators that can induce greater target gene expression. In addition, we also have constructed a modified regulator which can repress gene expression. The versatility of our system should prove useful for many applications in biology and gene therapy. PMID- 10837600 TI - Manufacturing and quality control of plasmid-based gene expression systems. AB - DNA plasmid-based gene expression systems are being widely investigated for the potential treatment of genetic and acquired disease and for DNA-based vaccination. A number of human clinical trials are in progress using plasmid based drugs. The regulatory framework that has been applied to biologicals such as recombinant DNA-derived proteins has proven to be generally applicable for regulating plasmid-based drugs as well. This was recently emphasized by the inclusion of therapeutic DNA plasmid products in the U.S. Food and Drug Administration's list of well-characterized biotechnology products. Present techniques for manufacturing and characterizing plasmids have been adapted from large-scale protein purification and from traditional molecular biology. Production of multi-gram quantities of plasmid, at purities of 95% or more, is currently possible, but further development of both manufacturing and analytical techniques is required. This review describes the approaches and methods currently used to manufacture and characterize DNA plasmids for pharmaceutical use, as well as recent changes in the regulatory environment that will impact future development and marketing of plasmids as human drugs. PMID- 10837601 TI - Interpolyelectrolyte and block ionomer complexes for gene delivery: physico chemical aspects. AB - Interpolyelectrolyte complexes formed as a result of the polyion coupling reaction between polynucleotide and polycation chains have been used for gene delivery. A general disadvantage of these systems is their reduced solubility. To overcome this problem, cationic copolymers comprised of polycation and poly(ethylene oxide) segments have been developed. These copolymers form block ionomer complexes with DNA that are water soluble even if all electrostatic charges are neutralized. This paper discusses physico-chemical aspects of formation and behavior of interpolyelectrolyte and block ionomer complexes and presents recent results on the use of block ionomer complexes for gene delivery. PMID- 10837602 TI - Chemistry and cellular aspects of cationic facial amphiphiles. AB - A series of compounds containing a bile acid core and a polyamine side chain have been synthesized to evaluate their ability to promote the uptake of DNA into cells. These compounds differ from conventional cationic lipids because they contain a positively charged chain attached to a facial amphiphile rather than to a hydrophobic moiety. Formulations of several of the designed compounds were found to dramatically increase the cellular uptake of both plasmid and oligonucleotide DNA. Moreover, initial experiments have shown that some of the compounds promote plasmid gene expression in various tissues after introduction into animals. These results have provided interesting information about structure activity relationships as well as clues to the mechanism of action that may lead to further improvements in the design. PMID- 10837603 TI - Recent advances in liposome technologies and their applications for systemic gene delivery. AB - The recent clinical successes experienced by liposomal drug delivery systems stem from the ability to produce well-defined liposomes that can be composed of a wide variety of lipids, have high drug-trapping efficiencies and have a narrow size distribution, averaging less than 100 nm in diameter. Agents that prolong the circulation lifetime of liposomes, enhance the delivery of liposomal drugs to specific target cells, or enhance the ability of liposomes to deliver drugs intracellularly can be incorporated to further increase the therapeutic activity. The physical and chemical requirements for optimum liposome drug delivery systems will likely apply to lipid-based gene delivery systems. As a result, the development of liposomal delivery systems for systemic gene delivery should follow similar strategies. PMID- 10837604 TI - Gene transfer with lipospermines and polyethylenimines. AB - It is an obvious and basic principle that to be efficient, gene therapy requires effective gene transfer followed by adequate gene expression. However, getting DNA, a pro-drug, into the cell and into the nucleus, remains a crucially limiting factor. Even recombinant viral methods still show poor performances in clinical situations and non-viral methods are considered classically to be of yet lower efficiency. Here, we consider the mode of action, the nature of the complexes formed with DNA and the transfection potentials of two categories of inert, cationic vectors, the lipospermines and polyethylenimine. Both are among the best vectors currently available for in vitro work. Moreover, polyethylenimine is proving to be a versatile and effective carrier for different in vivo situations, especially for delivering genes into the mammalian brain. PMID- 10837605 TI - Polylysine-based transfection systems utilizing receptor-mediated delivery. AB - Receptor-mediated gene transfer is a promising gene delivery technique. It employs a DNA-binding polycation, such as polylysine, to compact plasmid DNA to a size that can be taken up by cells (<100-200 nm). To allow internalization by receptor-mediated endocytosis, cell binding ligands, such as asialoglycoproteins or galactose for hepatocytes, anti-CD3 and anti-CD5 for T-cells, and transferrin, have been covalently attached to polylysine. Intracellular barriers for successful gene transfer include release of DNA complexes from endosomes or lysosomes, nuclear import of DNA complexes, and disassembly of the DNA-polylysine particles. Release of particles from internal vesicles has been achieved by the addition of lysosomotropic agents or glycerol to the transfection medium, or by the incorporation of endosomolytic compounds, such as viruses or membrane active peptides. This technique has already been used to transfect certain organs in vivo, including liver and lung. PMID- 10837606 TI - Synthetic peptide-based DNA complexes for nonviral gene delivery. AB - A major advantage of synthetic peptide-based DNA delivery systems is its flexibility. By design, the composition of the final complex can be easily modified in response to experimental results in vitro and in vivo to take advantage of specific peptide sequences to overcome extra- and intracellular barriers to gene delivery. The extreme heterogeneity which greatly complicates both the kinetics of DNA-poly(L-lysine) interaction and the thermodynamic stability of the final DNA complexes is avoided. Other unique features include the absence of biohazards related to the viral genome as well as the production of the viral vector and the absence of limitations on the size of the therapeutic genes that can be inserted in the recombinant viral vector. In principle, if the gene can be cloned into an expression plasmid, it can be delivered as a synthetic DNA complex. Since these synthetic delivery systems are composed of small peptides which may be poorly antigenic, they hold the promise of repeated gene administration, a highly desirable feature which will be important for gene targeting in vivo to endothelial cells, monocytes, hepatocytes and tumor cells. PMID- 10837607 TI - Pharmacokinetic considerations in somatic gene therapy. AB - It is anticipated that gene therapies will be useful for achieving long durations of action with little temporal fluctuation in the level of the therapeutic gene product, localized effects in specific tissues or cell types, and levels of biological products that can be regulated over time by drugs or physiological events. The effective clinical application of gene therapies will require detailed consideration of the pharmacokinetics of the gene and its gene product. This requires understanding not only the apparent kinetics of the bioactive gene product, but the intrinsic kinetics of each step involved in gene delivery, gene expression and the bioavailability of the gene product. Numerical models are described for three different approaches to gene therapy: (i) those that involve permanent integration of a transgene into the target cell, (ii) those that involve transient residence of the transgene within the cell, and (iii) those that allow control over gene expression by regulatory factors or administered drugs. Experimental studies describing the dynamics and kinetics of DNA in vivo and early pharmacokinetic studies of viral and non-viral systems are reviewed. PMID- 10837608 TI - Plasmid delivery to muscle: Recent advances in polymer delivery systems. AB - Preclinical studies involving intramuscular injection of plasmid into animals have revealed at least four significant variables that effect levels of gene expression (i.e., >fivefold effect over controls), including the formulation, injection technique, species and pretreatment of the muscle with myotoxic agents to induce muscle damage. The uptake of plasmid formulated in saline has been shown to be a saturable process, most likely via a receptor-mediated event involving the T tubules and caveolae. Pharmacokinetic studies have demonstrated that the bioavailability of injected plasmid to muscle cells is very low, due to rapid and extensive plasmid degradation by extracellular nucleases. We have developed protective, interactive, non-condensing (PINC) delivery systems designed to complex plasmids and to (i) protect plasmids from rapid nuclease degradation, (ii) disperse and retain intact plasmid in the muscle and (iii) facilitate the uptake of plasmid by muscle cells. PINC systems result in up to at least a one log increase in both the extent and levels of gene expression over plasmid formulated in saline. We have combined the PINC delivery systems with two different muscle-specific expression plasmids. After direct intramuscular injection of these gene medicines, we have shown both local myotrophic and neurotrophic effects of expressed human insulin-like growth factor (hIGF-I) and the secretion of biologically active human growth hormone (hGH) into the systemic circulation. PMID- 10837609 TI - Airway delivery of cationic lipid: DNA complexes for cystic fibrosis. AB - Cationic lipids are under active consideration as gene delivery vehicles for cystic fibrosis. Initial studies have shown cationic lipids to be effective agents of gene transfer to epithelial cells in vitro. Instillation of these vectors into animal models has led to widely different degrees of transfection of the airway epithelia. Newer generations of cationic lipids, with dramatically improved transfection efficiencies, appear to result in mostly alveolar gene delivery. Aerosol delivery of cationic lipid-DNA complexes has resulted in variable transfection of the airways in animal models. Initial human clinical trials using intranasal instillation have shown variable low levels of expression, accompanied by little toxicity. Recent developments in the formulation of cationic lipid-DNA complexes have resulted in an ability to aerosolize high concentrations of the complex, and should permit an evaluation of the efficacy of these delivery vehicles when aerosolized into cystic fibrosis patients. PMID- 10837610 TI - Arterial gene transfer of naked DNA for therapeutic angiogenesis: early clinical results. AB - Patients with critical limb ischemia constitute a potential target population for therapeutic angiogenesis. Because the growth of new collateral vessels can be achieved in a time interval of 1 month or less, these patients are suitable candidates for treatment with non-viral vectors intended to yield short-term gene expression. Accordingly, we applied naked plasmid DNA encoding for vascular endothelial growth factor, a secreted endothelial cell mitogen, to the hydrogel polymer coating of an angioplasty balloon. The balloon was then used to perform arterial gene transfer to the arterial circulation of the ischemic lower extremity. Using a dose-escalating strategy, it was possible to document that naked DNA was sufficient to generate evidence of new collateral growth by both magnetic resonance angiography and contrast angiography in the affected limb. These findings establish that the use of naked DNA may be suitable for gene therapy when the gene product is actively secreted from transfected cells. PMID- 10837611 TI - Pre-clinical trials using hepatic gene delivery. AB - The development of gene therapy as a potential technique for treating serious metabolic or infectious disorders has generated much interest. The general applicability of gene therapy depends on the efficient transfer of the desired gene to specific tissues and cells. One of the most attractive sites for gene transfer is the liver because it plays a major role in many metabolic processes and is involved in a large variety of diseases. Nonviral strategies have been conceived for delivering genes to the liver but this approach is still at the preclinical stage. This review outlines the more commonly used approaches and discusses the progress that has been made toward developing a widely applicable, clinically relevant gene transfer procedure for the liver. PMID- 10837612 TI - Cystic fibrosis clinical trials. AB - The ion transport abnormalities in cystic fibrosis are becoming increasingly well defined, although how these lead to lung pathology is still speculation. Correction of these defects could theoretically be achieved either through pharmacological means or via gene therapy. Pharmacological approaches include increasing the amount of CFTR protein that reaches its correct localisation in epithelial cells. Secondly, approaches have been suggested which could increase the function of the protein already present at this correct localisation. Finally, it may be possible to identify alternative channels which could subserve the function of CFTR. Gene therapy is theoretically an attractive proposition as it should circumvent each of the identified abnormalities in cystic fibrosis. The principal difficulty at present relates to delivering sufficient copies of the normal CFTR gene into the appropriate cell population in vivo. A number of clinical trials have now been undertaken and steady and encouraging progress has been made in moving this approach from theory to practice. PMID- 10837614 TI - Preface. PMID- 10837613 TI - Preclinical and clinical study of HER-2/neu-targeting cancer gene therapy. AB - Cationic liposomes have been used in many gene therapy approaches. The advantages of low toxicity, lack of immunological response, and easy preparation using cationic liposomes make multiple administrations possible, which may overcome the disadvantage of low transfection efficiency. Cationic liposomes, therefore, provide a promising procedure for delivering a therapeutic gene into cancer patients. Amplification or overexpression of the HER-2/neu oncogene is frequently found in breast and ovarian cancers and correlates with poor clinical prognosis. We have found that the adenovirus 5 E1A and the nontransforming simian virus 40 (SV40) large-T antigen mutant can inhibit HER-2/neu overexpression and reverse the HER-2/neu-mediated malignant phenotypes. By using the cationic liposome 3beta[N-(N',N'dimethylamino)ethanecarbamoyl]-cholesterol (DC-Chol), we successfully transferred E1A and/or large-T mutant into established orthotopic breast and ovarian cancer models. The survival of a treated group of mice was significantly prolonged and the expression of HER-2/neu oncogene was down regulated in vivo. A subsequent toxicity assay indicated that no significant toxicity was associated with the liposome-DNA complex administration even when we used ten times the dose needed to achieve a therapeutic effect. Based on these data, a phase I clinical trial of DC-Chol-mediated E1A gene therapy for ovarian and breast cancers that overexpress HER-2/neu has been initiated in our institute. In this article, we will review the development of HER-2/neu-targeting gene therapy using cationic liposomes. PMID- 10837615 TI - Creating and engineering human antibodies for immunotherapy. AB - Targeting in immunotherapy has traditionally been achieved by using monoclonal rodent antibodies. Despite gene-engineering, there are many problems and limitations associated with the non-human origin, the targeting specificity and the binding strength of these molecules. Now these issues may be addressed in a more rational way, by designing and then shaping, in vitro, the desired human antibodies. This review addresses how this may be achieved by the selection of monoclonal human antibodies from phage display libraries and the engineering of affinity and specificity thereafter. Phage display of antibody fragments has allowed access to large collections of different phage antibodies, created by cloning antibody V-genes from B-cells. Antibodies against any type of antigen may be derived from such repertoires, by rounds of enrichment on antigen and re amplification. This review presents the state of the art in rational antibody design and creation. It will highlight the strengths of this increasingly important field, which will aid in the generation of tailor-made targeting entities for immunotherapy. PMID- 10837616 TI - Production and selection of antigen-specific fully human monoclonal antibodies from mice engineered with human Ig loci. AB - The ability to produce highly specific fully human monoclonal antibodies to human antigens has potential significant applications to human therapy. This review describes the creation of novel mouse strains engineered to produce a diverse repertoire of fully human antibodies in the absence of mouse antibodies. These mouse strains have been generated by introducing megabase-sized human immunoglobulin loci, containing the majority of the human antibody gene repertoire, in nearly germline configuration, into mice deficient in mouse antibody production. The mice produce high levels of human IgMkappa and IgGkappa antibodies with a diverse adult-like repertoire. Upon immunization with multiple human antigens the mice generate high affinity, antigen-specific fully human monoclonal antibodies with neutralization activity. Comparison of these mice to other strains containing limited human antibody gene repertoire underscores the importance of the large number of variable genes for faithful reproduction of functional and diverse human antibody response in mice. PMID- 10837617 TI - Affinity reagents against tumour-associated extracellular molecules and newforming vessels. AB - Here we report some recent results of tumour targeting using extracellular matrix components of tumour stroma as targets. The possibility of using human recombinant antibodies in tumour targeting is also described. Preliminary results indicate that neovasculature markers can be targeted by recombinant antibodies and that they allow long residence time in tumours, thus exploiting the avidity properties of multivalent recombinant fragments. PMID- 10837618 TI - Immunotoxins for targeted cancer therapy. AB - Immunotoxins constitute a new modality for the treatment of cancer, since they target cells displaying specific surface-receptors or antigens. Immunotoxins contain a ligand such as a growth factor, monoclonal antibody, or fragment of an antibody which is connected to a protein toxin. After the ligand subunit binds to the surface of the target cell, the molecule internalizes and the toxin kills the cell. Bacterial toxins which have been targeted to cancer cells include Pseudomonas exotoxin and diphtheria toxin, which are well suited to forming recombinant single-chain or double-chain fusion toxins. Plant toxins include ricin, abrin, pokeweed antiviral protein, saporin and gelonin, and have generally been connected to ligands by disulfide-bond chemistry. Immunotoxins have been produced to target hematologic malignancies and solid tumors via a wide variety of growth factor receptors and antigens. Challenges facing the clinical application of immunotoxins are discussed. PMID- 10837619 TI - Targeted delivery of chemotherapeutics: tumor-activated prodrug therapy. AB - The potential of targeted delivery of chemotherapeutic drugs for the treatment of cancer has not yet been realized owing to the difficulty of delivering therapeutic concentrations to the target site. While in vivo studies in animal tumor models have produced very encouraging results, clinical studies with antibody-drug conjugates have been less successful. This paper will review the current status of the targeted delivery approach and analyze some of the reasons for the lack of success so far. Starting with a historical perspective, this review will end with a description of newer, more potent and specific antibody drug conjugates, which behave like tumor-activated prodrugs that may yet fulfil the promise of the targeted delivery approach for the treatment of cancer. PMID- 10837620 TI - Bispecific antibodies for treatment of cancer in experimental animal models and man. AB - Immunotherapy is a powerful anti-cancer treatment modality. However, despite numerous encouraging results obtained in pre-clinical studies, a definite breakthrough towards an established clinical treatment modality has as yet not occurred. Antibodies against tumor antigens have been shown to localise at the site of the tumor, but inadequate triggering of immune effector mechanisms have thwarted clinical efficacy thus far. Cellular immunotherapy has been hampered by limitations such as lack of specificity, down-regulation of major histocompatibility complex (MHC)-expression or Fas ligand up-regulation on tumor cells. This review focuses on the use of bispecific antibodies (BsAbs) for immunotherapy of cancer. Using BsAbs, it is possible to take advantage of the highly specific binding characteristics of antibodies and combine these with the powerful effector functions of cytotoxic immune effector cells. BsAbs share two different, monoclonal antibody-derived, antigen-recognizing moieties within one molecule. By dual binding, BsAbs reactive with a trigger molecule on an immune effector cell on the one hand and a surface antigen on a tumor target cell on the other are thus able to functionally focus the lytic activity of the immune effector cell towards the target cell. Over the last few years, the concept of BsAb-mediated tumor cell killing has been studied extensively both in preclinical models and in a number of phase I clinical trials. Promising pre-clinical results have been reported using tumor models in which diverse immune effector cell populations have been used. Despite this pre-clinical in vivo efficacy, the first clinical trials indicate that we are still not in a position to successfully treat human malignancies. This review discusses the production of BsAbs, the choice of trigger molecules in combination with potential effector cells and the preclinical models that have led to the current use of BsAbs in experimental clinical trials. It has become clear that appropriate immune cell activation and establishing a favourable effector-to-target cell ratio will have direct impact on the efficacy of the therapeutic approaches using BsAbs. New directions are discussed, i.e. finding appropriate dosage schemes by which immune effector cells become redirected without inducing hyporesponsiveness, defining possibilities for combining different immune effector cell populations and creating an in situ tumor environment that allows maximal tumoricidal activity PMID- 10837621 TI - Man-made superantigens: Tumor-selective agents for T-cell-based therapy. AB - Superantigens (SAgs) are a collection of bacterial and viral proteins with potent immunostimulatory properties. SAgs bind to Major Histocompatibility Complex Class II (MHC II) molecules of antigen presenting cells (APCs) and activate a high frequency of T lymphocytes. To target a T-cell attack against tumor cells we genetically linked tumor-specific antibody Fab fragments to the SAg Staphylococcal enterotoxin A (SEA). Fab-SEA fusion protein efficiently targeted to solid tumors and induced a T-cell-mediated eradication of established metastases in animal models. Successful therapy was T-cell-dependent and required tumor specificity of the Fab moiety of the Fab-SEA fusion protein. Due to the high affinity of SAg for MHC II, a limitation of this approach was retention of Fab-SEA proteins in normal tissues expressing MHC II, which caused systemic immune activation and dose limiting toxicity. We recently solved the structure of SEA and applied structure-based drug design to develop a novel generation of 'man made' SAg with improved pharmacological and pharmacokinetic properties. Mutation of the major MHC II binding site of SEA substantially reduced retention in MHC II(+) tissues and systemic toxicity, while local immune activation at targeted tumor sites was retained. The Fab-SEA mutants display a 10000-fold higher affinity for tumor tissue compared to normal tissue and the therapeutic window was improved >100-fold compared to native Fab-SEA protein. Thus protein engineering can be applied to convert harmful bacterial toxins into tolerable tumor-specific agents. PMID- 10837622 TI - Overview of clinical trials employing antibody-targeted superantigens. AB - Two Phase I clinical trials have been conducted using PNU-214565, a recombinant fusion protein of C242Fab and staphylococcal enterotoxin A (SEA). The initial escalating single dose trial was performed to determine safety and define toxicities. Cumulative doses determined to be safe as single doses were incorporated in an escalating, repeated dose regimen. Twenty-one patients were treated in the single dose and 27 in the repeated dose trials. Patient demographics were equivalent in both, as were the toxicities encountered - primarily fever and hypotension. Three patients in the single dose regimen treated at 0.5 ng/kg experienced grade 3 fever and/or hypotension, and one patient in the repeated dose trial had a dose-limiting grade 4 hypotension (2.75 ng/kg). TNFalpha and IL-2 induction in circulating blood preceded the development of clinical symptoms. One partial response was observed in the repeated dose trial. Pre-existing anti-SEA plasma antibodies protect patients against toxicity at a given drug dose. Based on these findings, a pharmacodynamically-based dosing scheme is currently being tested in a new repeated dose trial. PMID- 10837623 TI - Antibodies for targeted gene therapy: extracellular gene targeting and intracellular expression. AB - Antibody genes of human origin and human antibodies directed against human proteins have become widely available in recent years. These are valuable reagents for gene therapy applications, in which the use of human proteins and genes allows for increased therapeutic benefit. Engineered human antibodies can be used in gene therapy both as a component of a gene delivery system and as a therapeutic gene. As the targeting moiety of a gene delivery system, the antibody should meet certain criteria that have been previously determined from other clinical applications of antibodies. These include bioavailability, specificity for the target cell, and rapid clearance. In addition, if repeat delivery of therapeutic genes is going to be needed, then gene delivery vectors should be non immunogenic to allow repeated administration. The use of human antibodies in this application should therefore be superior to approaches which use rodent-derived antibodies. Another application of antibodies in gene therapy is the use of antibodies expressed inside the cell (intrabodies) as therapeutic agents. The power of the immune system to rearrange a limited set of genes to create recognition sites for any known molecule is well documented. The ability to harness this information and use these highly specific binding molecules as medicines to inhibit an unwanted cellular function is a promising advance in the field of molecular medicine, and in particular, in the field of intracellular immunization. PMID- 10837624 TI - Tyrosine kinase chimeras for antigen-selective T-body therapy. AB - Protein tyrosine kinases (PTKs) transmit activation signals in almost every cell type, including immune effector cells. The aberrant or constitutive activation of PTKs can often cause neoplastic transformation. The use of chimeric receptors based on PTKs may enable us to elucidate the signaling pathways of normal immune cells and other cell types, and the abnormal events that can lead to malignant transformation. In this review, we focus on antigen specific chimeric PTKs in which antibody-derived scFv are joined to the Syk family of PTKs. These chimeric receptors yielded reagents that can selectively redirect immune effector cells and specifically activate them to produce cytokines or lyse their target. The advantages of using such PTK-based chimeras to redirect lymphocytes to tumor targets and their potential as an immunotherapeutic approach to malignant disease is discussed. PMID- 10837625 TI - Protein release from polyphosphazene matrices. AB - Polyphosphazenes have been exploited as carriers for protein delivery due to versatility of molecular structures and sophisticated spectrum of chemical and physical properties. Ease of structural manipulations for this class of organometallic polymers allows efficient control over physico-chemical parameters of polyphosphazenes including their biodegradability and matrix permeability. Some polyphosphazenes offer additional advantages as protein delivery vehicles since microencapsulation of substrates in these systems can be achieved under remarkably mild physiological conditions. Because of these properties polyphosphazenes have tremendous potential as matrices for protein release as shown by studies both in vitro and in vivo. PMID- 10837626 TI - Temperature-responsive gels and thermogelling polymer matrices for protein and peptide delivery. AB - The explosive development of protein and peptide drugs has ushered in a great need for effective delivery systems for such drugs. Two decades of studies of protein delivery from polymeric systems has revealed the great potential of gels that respond to environmental stimuli, such as temperature, pH, etc. Protein and peptide release can be engineered to occur in a pulsatile mode. The mechanism of this release is greatly affected by polymer design. 'Intelligent' amphiphilic copolymers emerge as a novel trend in the application of thermodynamically stable self-assembling lyophilic colloids to protein and peptide delivery. PMID- 10837627 TI - Inclusion and release of proteins from polysaccharide-based polyion complexes. AB - The notion of a polyelectrolyte complex is well established for the complexation of two polymers one anionic, the other cationic. Electronic microscopy studies have shown the formation of a fibrilar structure. A method for the preparation of polyionic hydrogels from the complexation of chitosan and xanthan is reported. Electronic microscopy studies have shown the formation of a fibrilar structure. Stable hydrogels have been used to immobilize xylanase, lipase and protease. The immobilized xylanase and lipase activity was significantly higher than that of the free enzyme. PMID- 10837628 TI - Protein release from collagen matrices. AB - The effective delivery of protein drugs is an important research subject in the field of pharmacology, and to prolong the effect of protein drugs, many studies are being conducted to control the release of proteins from various carrier materials. Collagen is one of the most useful candidates for this purpose, and many studies have been reported; pharmaceutical formulations containing collagen in gel, film and sponge form are used to incorporate low-molecular-weight compounds such as antibiotics and carcinostatics, and the release of these compounds is controlled by the concentration of the gel as well as the shape and degree of crosslinking of the matrix. However, it is still difficult to retain protein drugs in the collagen. In this article, we report on the controlled release of protein drugs using collagen which exhibits good biocompatibility as a carrier, focusing on a new drug delivery system, the Minipellet, which we have developed. PMID- 10837629 TI - Protein release from alginate matrices. AB - There are a variety of both natural and synthetic polymeric systems that have been investigated for the controlled release of proteins. Many of the procedures employed to incorporate proteins into a polymeric matrix can be harsh and often cause denaturation of the active agent. Alginate, a naturally occurring biopolymer extracted from brown algae (kelp), has several unique properties that have enabled it to be used as a matrix for the entrapment and/or delivery of a variety of biological agents. Alginate polymers are a family of linear unbranched polysaccharides which contain varying amounts of 1,4'-linked beta-D-mannuronic acid and alpha-L-guluronic acid residues. The residues may vary widely in composition and sequence and are arranged in a pattern of blocks along the chain. Alginate can be ionically crosslinked by the addition of divalent cations in aqueous solution. The relatively mild gelation process has enabled not only proteins, but cells and DNA to be incorporated into alginate matrices with retention of full biological activity. Furthermore, by selection of the type of alginate and coating agent, the pore size, degradation rate, and ultimately release kinetics can be controlled. Gels of different morphologies can be prepared including large block matrices, large beads (>1 mm in diameter) and microbeads (<0.2 mm in diameter). In situ gelling systems have also been made by the application of alginate to the cornea, or on the surfaces of wounds. Alginate is a bioadhesive polymer which can be advantageous for the site specific delivery to mucosal tissues. All of these properties, in addition to the nonimmunogenicity of alginate, have led to an increased use of this polymer as a protein delivery system. This review will discuss the chemistry of alginate, its gelation mechanisms, and the physical properties of alginate gels. Emphasis will be placed on applications in which biomolecules have been incorporated into and released from alginate systems. PMID- 10837630 TI - Protein release from gelatin matrices. AB - Gelatin is a denatured, biodegradable protein obtained by acid and alkaline processing of collagen. This processing affects the electrical nature of collagen, yielding gelatin with different isoelectric points (IEPs). When mixed with positively or negatively charged gelatin, an oppositely charged protein will ionically interact to form a polyion complex. This review article describes protein release from charged gelatin matrices on the basis of this polyion complexation. The biodegradable hydrogel matrices are prepared by chemical crosslinking of acidic or basic gelatin and are enzymatically degraded in the body with time. The degradation is controllable by changing the extent of crosslinking, which, in turn, produces hydrogels with different water contents. The time course of protein release is in good accordance with the rate of hydrogel degradation. It is very likely that the protein drug complexed with gelatin hydrogel is released as a result of its biodegradation. This gelatin hydrogel system releases the protein drug under maintenance of biological activity. This article will focus on experimental data that sustained release of growth factor from the gelatin hydrogels is very effective in exerting the biological functions of the growth factor. PMID- 10837631 TI - Sustained release emphasizing recombinant human bone morphogenetic protein-2. AB - Bone homeostasis is a dynamic process involving a myriad of cells and substrates modulated by regulatory signals such as hormones, growth and differentiating factors. When this environment is damaged, the regenerative sequalae follows a programmed pattern, and the capacity for successful recovery is often dependent on the extent of the injury. Many bony deficits that are excessively traumatic will not result in complete recovery and require therapeutic intervention(s) such as autografting or grafting from banked bone. However, for numerous reasons, an unacceptably high rate of failure is associated with these conventional therapies. Thus, alternative approaches are under investigation. A class of osteogenic regulatory molecules, the bone morphogenetic proteins (BMPs), have been isolated, cloned and characterized as potent supplements to augment bone regeneration. Optimizing a therapeutic application for BMPs may be dependent upon localized sustained release which in kind relies on a safe and well characterized carrier system. This review will discuss the current status of BMPs in bone regeneration and specifically will present the potential for a clinical therapeutic role of recombinant human BMP-2 sustained release carrier systems. PMID- 10837632 TI - Interactions of liposomes and lipid-based carrier systems with blood proteins: Relation to clearance behaviour in vivo. AB - Liposomes and lipid-based drug delivery systems have been used extensively over the last decade to improve the pharmacological and therapeutic activity of a wide variety of drugs. More recently, this class of carrier systems has been used for the delivery of relatively large DNA and RNA-based drugs, including plasmids, antisense oligonucleotides and ribozymes. Despite recent successes in prolonging the circulation times of liposomes, virtually all lipid compositions studied to date are removed from the plasma compartment within 24h after administration by the cells and tissues of the reticuloendothelial system (RES). Plasma proteins have long been thought to play a critical role in this process but only a few efforts were made to evaluate the relevant importance of plasma protein-liposome interactions in the clearance process. Strategies to increase the bioavailability of liposomal drugs have included altering lipid compositions and charge, increasing lipid doses, and incorporating surface coatings. All of these modifications can influence membrane-protein interactions. In this article, we will focus on our experiences with liposome-blood protein interactions and how alterations in the chemical and physical properties of the carrier system influence the interactions with blood proteins and circulation times. PMID- 10837633 TI - The complement system in liposome clearance: Can complement deposition be inhibited? AB - The activation of complement results in the opsonization of particles for removal by the reticuloendothelial system. Experimental evidence suggests that complement mediated clearance of liposomal systems may significantly contribute to liposome biodistribution. Because of the multiplicity of complement activation mechanisms and the large number of proteins in the pathway, there are multiple opportunities to reduce or eliminate the opsonic effects of complement activation. This review addresses the state of our understanding of the interaction of liposomes with complement proteins and suggests some approaches to minimize complement activation. PMID- 10837634 TI - Structural and metabolic consequences of liposome-lipoprotein interactions. AB - The two major proposed uses for liposomes, i.e., drug delivery and mobilization of peripheral deposits of cholesterol, each impose requirements and restrictions on liposomal structure, particularly as it affects interactions with lipoproteins. This chapter focuses on the role of lipoproteins and apolipoproteins in (1) disrupting membrane structure and causing the leakage of liposomal contents by inducing disc formation and (2) marking liposomes for whole particle uptake by receptors involved in lipoprotein metabolism. Control of membrane stability and whole-particle half-life can be achieved by several strategies, such as membrane stiffening, shielding the membrane surface, and increasing the dose or predosing with "empty" liposomes. The rationales and applicabilities of these strategies are discussed in the contexts of liposomes as drug delivery vehicles and as antiatherogenic particles. Directions for further basic and applied research are also presented. PMID- 10837635 TI - Serum-mediated recognition of liposomes by phagocytic cells of the reticuloendothelial system - The concept of tissue specificity. AB - The mechanisms responsible for the clearance and recognition of intravenously injected liposomes by phagocytic cells of the reticuloendothelial system are presently not well understood. Here, we address a multiplicity of physicochemical and physiopathological factors which influence the clearance kinetics and tissue distribution of liposomes administered into the circulation. Emphasis is particularly drawn towards the concept of liposome-blood protein interaction and its role in liposome recognition by various phagocytes of the reticuloendothelial system. PMID- 10837636 TI - Identification of proteins mediating clearance of liposomes using a liver perfusion system. AB - The objective of this paper is to identify the principal blood components governing the fate of liposomes in blood circulation. Information based on an isolated perfused liver system in rats has revealed the central role of the complement system in enhancing the uptake of liposomes by the liver. A species difference was an important factor in determining the uptake mechanisms of liposomes by the liver. Limited evidence revealed the tendency that opsonin dependent hepatic uptake is principal in rats, while opsonin-independent or dysopsonin-dependent uptake governs in mice, although there are some exceptions. These studies provide us with important information for understanding the uptake mechanisms of liposomes by the liver, and useful insights in predicting the in vivo disposition of liposomes in humans. PMID- 10837637 TI - Receptor versus non-receptor mediated clearance of liposomes. AB - Numerous studies have appeared over the years dealing with liposome-cell interaction mechanisms, most of them performed under in vitro conditions with isolated cell populations or cell lines. It is remarkable that, nonetheless, there hardly seem to exist established and generally accepted views on how precisely liposomes interact with cells and by what parameters this is influenced. In this article we will summarize and discuss the most relevant studies (in our opinion) on this matter in relation to in vivo conditions and with special attention to the relation between scavenger, complement and PS receptors.Researchers in the field have long been aware of the interaction of liposomes with blood proteins and their potential involvement in the process of liposome elimination from the blood circulation. A few of these 'opsonizing' proteins have been identified, but it is not clear to what extent each of them determines the fate of the liposome in the blood stream and how liposomal parameters such as size, charge and rigidity play a role in this process. We will include in this article our own recent observations on a thus far largely ignored class of such liposomal 'opsonins', the apolipoproteins. This class of plasma proteins, which physiologically are instrumental in hepatic lipoprotein clearance and processing, has been shown to contribute specifically to hepatocyte-mediated uptake of liposomes.Separately, as opposed to the fate of plain liposomes, we briefly touch on the clearance of surface-modified liposomes, which are designed to actively target specific cells or tissues. Plasma proteins are not usually supposed to play a significant role in the clearance of such liposomes. We will summarize these studies and address in this connection the question of how plasma proteins may interfere with such active targeting attempts. PMID- 10837638 TI - Clearance properties of liposomes involving conjugated proteins for targeting. AB - This review examines methods of protein conjugation onto liposomes and the effects of surface bound protein on the liposomes' biological behavior. It is evident that the presence of a conjugated protein significantly alters the attributes of targeted liposomes. Specifically, protein conjugation can result in dramatic increases in liposome size, enhanced immunogenicity, and increased plasma elimination. Techniques are discussed for preventing some of the physical (size) and biological (immunogenic) alterations involving the use of PEG-lipids and drug loaded liposomes. In addition, the advantages of conjugating antibodies via carbohydrate moieties, to minimize changes in antibody binding and tertiary structure as well as effectively decreasing plasma elimination, are also discussed. It is, however, apparent that the accessibility of targeted liposomes to extravascular sites is a key step that will require further study and it is, therefore, anticipated that with the development of novel ligands and novel ligand-liposome interactions, the therapeutic utility of targeting strategies will likely be realized. PMID- 10837639 TI - Theoretical considerations of RES-avoiding liposomes: Molecular mechanics and chemistry of liposome interactions. AB - The development of long-circulating, RES-avoiding liposomes has become a remarkable milestone in the progress of contemporary pharmacology. Drugs incorporated in such liposomes are protected from fast metabolization and clearance, and can be further targeted to a desired tissue site. Ideally, future developments should result in drug carriers which can identify and act upon their targets with even higher efficiency and selectivity, preferably close to or exceeding that of the natural immune cells.Further increasing carrier 'inertness' with regard to the normal biological milieu is the major requirement for future success. The ability of natural blood components to circulate with blood for several days and weeks presents both the motivation and the challenge for further research. Today, even the best available preparations are inferior to natural proteins and cells with regard to their ability to remain in circulation by approximately two orders of magnitude.In view of the above, it seems vitally important to determine the mechanisms responsible for glycolipid- or polymer modified liposome protection against RES, and whether any potentially useful mechanisms have been underutilized. Furthermore, identification of quantitative dependencies between liposome structure and pharmacokinetics (and mechanisms underlying such dependencies) would benefit future research and reduce the cost of development.This paper discusses the relationships between liposome structure and circulation with respect to the theoretical mechanistic models of mass transfer, liposome interactions with cells and blood proteins, and boundary effects resulting from surface modification. Special attention is paid to the practical application and limitations of the models. PMID- 10837641 TI - Preface. PMID- 10837640 TI - Controlling liposome blood clearance by surface-grafted polymers. AB - The incorporation of polymer-lipid conjugates, initially using PEG and subsequently other selected flexible, hydrophilic polymers, into lipid bilayers gives rise to sterically stabilized liposomes that exhibit reduced blood clearance and concomitant changes in tissue distribution largely because of reduced, but not eliminated, phagocytic uptake. Changes in tissue distribution includes 'passive' targeting localization into sites of tumors, infection, inflammation characterized by presence of a 'leaky' vasculature which represent useful applications for drug delivery. The polymer forms a surface coating which has been characterized by physical measurements and it appears to function through steric inhibition of the protein binding and cellular interactions leading to phagocytic uptake. The current understanding of the physical and biological properties are reviewed. Ongoing work in the field involves interests to increase complexity such as addition of (1) selective targeting ligands by chemical conjugation to the exterior surface of the polymer coating, (2) capabilities for intracellular release of encapsulated agents into the cytoplasm, and (3) both simultaneously. PMID- 10837642 TI - Aluminum compounds as vaccine adjuvants. AB - Aluminum compounds are the only adjuvants used widely with routine human vaccines and are the most common adjuvants in veterinary vaccines also. Though there has been a search for alternate adjuvants, aluminum adjuvants will continue to be used for many years due to their good track record of safety, low cost and adjuvanticity with a variety of antigens. For infections that can be prevented by induction of serum antibodies, aluminum adjuvants formulated under optimal conditions are the adjuvants of choice. It is important to select carefully the type of aluminum adjuvant and optimize the conditions of adsorption for every antigen since this process is dependent upon the physico-chemical characteristics of both the antigens and aluminum adjuvants. Adsorption of antigens onto aluminum compounds depends heavily on electrostatic forces between adjuvant and antigen. Two commonly used aluminum adjuvants, aluminum hydroxide and aluminum phosphate have opposite charge at a neutral pH. The mechanism of adjuvanticity of aluminum compounds includes formation of a depot; efficient uptake of aluminum adsorbed antigen particles by antigen presenting cells due their particulate nature and optimal size (<10 um); and stimulation of immune competent cells of the body through activation of complement, induction of eosinophilia and activation of macrophages. Limitations of aluminum adjuvants include local reactions, augmentation of IgE antibody responses, ineffectiveness for some antigens and inability to augment cell-mediated immune responses, especially cytotoxic T-cell responses. PMID- 10837643 TI - Adjuvant activity of incomplete Freund's adjuvant. AB - The water-in-oil emulsion adjuvants developed by Jules Freund and coworkers have been, and continue to be, used in research and veterinary vaccines. Incomplete Freund's Adjuvant (IFA) has also been widely tested in humans where it was shown to augment antibody responses greater than other adjuvants, such as alum, and to be well tolerated. Toxicity associated with the use of IFA has been controlled by the use of high grade oils and purified surfactants and clinical testing is ongoing to day. One potential limitation of IFA is that it does not augment the types of cellular immune responses that thought to be critical to the control of many viral infections and tumors. However, IFA can be mixed with or used in conjunction with other adjuvant-active compounds, such as saponins and cytokines. These mixed formulations or vaccine protocols have proved successful in animal testing and may soon be tested in clinical trials. Thus, the toxicity and potential limited adjuvant activity of IFA can be controlled or corrected making IFA a useful product in the vaccine industry. PMID- 10837644 TI - Development and application of PROVAX adjuvant formulation for subunit cancer vaccines. AB - A major challenge facing the development of subunit vaccines comprised of well defined recombinant antigens is their weak immunogenicity and inability to induce effective cytotoxic T cell (CTL) responses. Adjuvants aimed at increasing the immunogenicity of recombinant antigens remain a focus in vaccine development. The potency of an adjuvant is linked to specific stimulation of T cell responses, involving TH1 and TH2 subsets of CD4(+) T helper cells and CD8(+) CTL and B cell mediated antibody responses. As a result of the existence of two distinct intra cellular pathways for antigen processing, immunization with exogenous antigens often shows a greater propensity for T helper and antibody responses, but not CD8(+) CTL responses. However, existing experimental evidence suggests that CD8(+) CTLs, which are critical in the elimination of viral-infected and neoplastic cells, can be elicited with soluble antigens when delivered in appropriate formulations or adjuvants. This review focuses on the properties of PROVAX adjuvant in inducing antigen-specific CTL responses, antibody responses and tumor regression in experimental models and its potential application for the development of recombinant cancer vaccines. PMID- 10837645 TI - Development of adjuvant-active nonionic block copolymers. AB - Nonionic block copolymers are surfactants synthesized using propylene oxide (PO) and ethylene oxide (EO) which are organized as 'blocks' of polyoxyethylene (POE) and polyoxypropylene (POP). These copolymers can be designed and synthesized using variable amounts of the PO and EO and with differential arrangement of the POP and POE blocks so that individual products have unique physicochemical properties. The copolymers that have been most thoroughly evaluated in vaccine research are linear with the polymer blocks organized as POE-POP-POE. Low molecular weight (MW) copolymers, 3-6 kDa, of this type have been used in oil based emulsion formulations, whereas high-MW copolymers, >9 kDa, can be used in aqueous formulations. The adjuvant activity of nonionic block copolymers is influenced greatly by the size of the POP core block. As the size of this block is increased so is the adjuvant activity of the copolymer; peak activity is achieved using copolymers with POP cores that are 12-15 kDa. However, adjuvant activity is also affected by the amount of POE with low concentrations, 5-10%, being optimal. The type of immune responses produced is also influenced by the POE content. Copolymers with 10% POE preferentially augment Type 2 helper T lymphocyte responses which support antibody responses, including mucosal antibody responses. Copolymers with <10% POE augment both Type 1 and Type 2 helper T lymphocyte responses, which support a broader range of antibody responses and cellular immune responses. This property may allow for vaccines to be 'customized' by using adjuvant-active nonionic block copolymers that will augment the most appropriate types of immune responses. PMID- 10837646 TI - Poly(lactide-co-glycolide) microparticles for the development of single-dose controlled-release vaccines. AB - With the exception of the provision of clean water supplies, vaccination remains the most successful public health intervention strategy for the control of infectious diseases. However, the logistics of delivering at least two to three doses of vaccines to achieve protective immunity are complex and compliance is frequently inadequate, particularly in developing countries. In addition, newly developed purified subunit and synthetic vaccines are often poorly immunogenic and need to be administered with potent vaccine adjuvants. Microparticles prepared from the biodegradable and biocompatible polymers, the poly(lactide-co glycolides) or (PLG), have been shown to be effective adjuvants for a number of antigens. Moreover, PLG microparticles can control the rate of release of entrapped antigens and therefore, offer potential for the development of single dose vaccines. To prepare single-dose vaccines, microparticles with different antigen release rates may be combined as a single formulation to mimic the timing of the administration of booster doses of vaccine. If necessary, adjuvants may also be entrapped within the microparticles or, alternatively, they may be co administered. The major problems which may restrict the development of microparticles as single-dose vaccines include the instability of vaccine antigens during microencapsulation, during storage of the microparticles and during hydration of the microparticles following in vivo administration. In the present review, we discuss the adjuvant effect of PLG microparticles, and also their potential for the development of single-dose vaccines through the use of controlled-release technology. PMID- 10837647 TI - ISCOMs and other saponin based adjuvants. AB - Saponins are chemically a heterogeneous group of sterol glycosides and triterpene glycosides which are common constituents of plants. One source of triterpenoid saponins obtained from the bark of Quillaia saponaria Molina (the soap bark tree) have been known to cause substantial enhancement of immune responses since the 1920s. Despite their use in animal vaccines, the development of saponin-based formulations for human vaccines has been impeded by their complexity and concerns about toxicity. This review briefly covers the use of saponins for animal vaccines but focuses mainly on the development of these adjuvants for use in man. Important aspects include preparation of purified or highly defined saponin fractions, improved understanding of the relationships between adjuvant activity, toxicity and structure of saponins and formulation of saponins into structures with reduced toxicity such as ISCOMs. Recent developments in the understanding of cellular interactions, cytokine induction and the in vivo localisation of saponin containing formulations will also be reviewed. PMID- 10837648 TI - Targeting immune response induction with cochleate and liposome-based vaccines. AB - The immune response generated by infection with a pathogenic organism, or by vaccination with a live attenuated or whole killed pathogen, often does not stimulate optimal protection against that organism. Lipid matrix-based subunit vaccines can be used to produce custom-designed vaccines, that elicit desired immune responses targeted to specific parts of the pathogen that are relevant to protection. Harmful or competitive responses can be minimized or avoided. Earlier work with liposomes has allowed the development of a new class of subunit vaccines called cochleate delivery vehicles, whose structure and properties are very different from liposomes. Protein and DNA cochleates are highly effective vaccines when given via mucosal or parenteral routes, including oral, intranasal, intramuscular, or subcutaneous. Strong, long-lasting, mucosal and circulating, antibody and cell-mediated responses are generated. Protection from challenge with live viruses following oral or intramuscular administration has been achieved. PMID- 10837649 TI - Cell-based tissue engineering therapies: the influence of whole body physiology. AB - A technology has been developed to isolate a developmentally and phenotypically homogeneous population of pluripotent human mesenchymal stem cells (hMSCs) from adult bone marrow and mitotically expand these cells in culture. These hMSCs have osteoblasts as one of their potential developmental end-stage phenotypes, and, in addition to their osteogenic potential, these hMSCs synthesize and secrete a variety of macromolecules that are known regulators of osteoclast differentiation and activity. In this review, data are presented that demonstrate the phenotypic and developmental homogeneity of the cells in hMSC cultures, as well as their ability to differentiate along multiple phenotypic pathways and serve as regulatory cells for hematopoietic and bone-resorbing cells. In addition, a logic and preliminary data are presented that support the use of hMSCs in the prevention and treatment of age-related and postmenopausal osteoporosis. Since hMSC differentiation and phenotypic expression are controlled by regulatory molecules synthesized and secreted by a variety of local and systemic mechanisms, the issue of whole organism physiology is addressed in considering tissue engineering logics. PMID- 10837650 TI - Culture of organized cell communities. AB - Cells cultured in vitro will tend to retain their differentiated phenotype under conditions that resemble their natural in vivo environment, for example, when cultured on polymer scaffolds in tissue culture bioreactors. In this chapter, we define organized cell communities as three-dimensional in vitro grown cell polymer constructs that display important structural and functional features of the natural tissue. We review representative studies in which the research goal was to culture organized cell communities resembling cartilage, bone, skeletal muscle or cardiac-like tissue. These constructs can potentially serve as tissue equivalents for in vivo transplantation or as a model system for the in vitro testing of cell and tissue-level responses to molecular, mechanical or genetic manipulations. PMID- 10837651 TI - Development of engineered cells for implantation in gene therapy. AB - Human gene therapy is based on the technology of genetic engineering of cells, either through ex vivo or in vivo methods of gene transfer. Many autologous cell types have been successfully modified to deliver recombinant gene products. An alternate form of gene therapy based on genetic modification of non-autologous cells is described. Protection within immuno-isolating devices would allow implantation of well-established recombinant cell lines in different allogeneic hosts, potentially offering a more cost-effective approach to gene therapy. Implantation with microencapsulated fibroblasts and myoblasts has resulted in successful recombinant product delivery in vivo. Correction of disease phenotypes in animal models of human genetic diseases has also been achieved. Cell types such as myoblasts which can differentiate terminally within the implantation device are particularly promising for the future development of this method of gene therapy. PMID- 10837652 TI - Development of engineered pancreatic beta-cell lines for cell therapy of diabetes. AB - Insulin-secreting pancreatic beta-cell lines represent a promising approach for treatment of insulin-dependent diabetes mellitus (IDDM). Our laboratory has developed a number of highly-differentiated beta-cell lines in transgenic mice. These cells produce insulin amounts comparable to normal pancreatic islets and release it in response to physiological insulin secretagogues. Using a reversible transformation system it has become possible to tightly regulate cell replication in these beta-cell lines both in culture and in vivo. By employing adenovirus genes which downreguate antigen presentation and increase cell resistance to cytokines mouse beta cells could be transplanted across allogeneic barriers. These approaches could be applied to the development of human beta-cell lines by genetic engineering of isolated human islets. PMID- 10837653 TI - Gene therapy for tissue repair and regeneration. AB - This review presents a current overview of the discipline of human gene therapy. In addition, a gene therapy method is described in which plasmid genes are transferred from a structural matrix carrier into fresh wound sites so as to enhance tissue repair and regeneration. The potential to develop a gene therapy for bone regeneration is discussed in detail. PMID- 10837654 TI - Materials for protein delivery in tissue engineering. AB - The ability of protein agents to modulate cellular behaviors, such as motility, proliferation, adhesion and function, is the subject of intense research; new therapies involving proteins will likely result. Unfortunately, many proteins have short half-lives and the potential for toxicity after systemic delivery, so traditional routes of administration are not appropriate. Alternate methods for sustained delivery of these agents to the desired cells and tissues in biologically active conformations and concentrations are necessary. Techniques similar to those long used in the controlled delivery of drugs have been used to administer certain growth factors to cells and tissues; although clinical success has been limited to date, studies in animal models suggest the potential for tremendous advances in the near future. This review outlines the basic technology of controlled protein delivery using polymeric materials, and discusses some of the techniques under investigation for the efficient administration of proteins in tissue engineering. PMID- 10837655 TI - Materials for immunoisolated cell transplantation. AB - Encapsulated cell therapy provides site-specific continuous delivery of cell synthesized molecules. Cell encapsulation therapy is based on the concept of immunoisolation. Foreign cells are surrounded with a semi-permeable membrane prior to transplantation to shield them from the host's natural defense system. This membrane is selectively permeable to transport of nutrients and therapeutic agents but relatively impermeable to larger molecules and cells of the hosts' immune system. Most encapsulation devices also utilize an internal matrix to keep cells suspended within the capsule. Proper choice of materials and materials processing techniques to formulate membrane and matrix components is essential to the success of these devices. A successful encapsulation device recreates the natural three-dimensional tissue environment that supports cell function and maintains cell viability. This review summarizes recent developments in materials development for cell encapsulation devices and highlights some ongoing challenges faced by those in the field. PMID- 10837656 TI - Host response to tissue engineered devices. AB - The two main components of a tissue engineered device are the transplanted cells and the biomaterial, creating a device for the restoration or modification of tissue or organ function. The implantation of polymer/cell constructs combines concepts of biomaterials and cell transplantation. The interconnections between the host responses to the biomaterial and transplanted cells determines the biocompatibility of the device. This review describes the inflammatory response to the biomaterial component and immune response towards transplanted cells. Emphasis is on how the presence of the transplanted cell construct affects the host response. The inflammatory response towards a biomaterial can impact the immune response towards transplanted cells and vice versa. Immune rejection is the most important host response towards the cellular component of tissue engineered devices containing allogeneic, xenogeneic or immunogenic ex vivo manipulated autologous cells. The immune mechanisms towards allografts and xenografts are outlined to provide a basis for the mechanistic hypotheses of the immune response towards encapsulated cells, with antigen shedding and the indirect pathway of antigen presentation predominating. A review of experimental evidence illustrates examples of the inflammatory response towards biodegradable polymer scaffold materials, examples of devices appropriately integrated as assessed morphologically with the host for various applications including bone, nerve, and skin regeneration, and of the immune response towards encapsulated allogeneic and xenogeneic cells. PMID- 10837657 TI - Biomechanics of cell interactions in shear fields. AB - Cellular interactions play a key role in diverse biological processes within the cardiovascular system. Targeting of leukocytes to sites of inflammation is viewed as a multistage process of sequential involvement of distinct adhesion molecules on the leukocyte and endothelial cell (EC) surface that is dictated by the local fluid dynamic environment. For neutrophils, the initial contact and rolling along the vessel wall are mediated primarily by selecting. Subsequent firm adhesion requires activation of neutrophil P, integrins and binding to their ligand ICAM-1 on the EC surface. The final step of this cascade of events includes neutrophil transmigration to extravascular tissue space. The neutrophil model of emigration in inflammation has been extended and refined to account for monocyte and T cell interactions with ECs. Platelet adhesion to thrombogenic surfaces (i.e. immobilized von Willebrand factor) under flow follows the general principles of leukocyte extravasation. More specifically, platelet glycoprotein (GP) Ib alpha appears to mediate an initial selectin-like tethering platelet-vWf interaction, followed by alpha II beta beta 3 integrin activation and firm adhesion. Some of the signaling mechanisms associated with cellular interactions in inflammatory and thrombotic processes are discussed. These basic principles are also discussed in the context of tissue engineering research. PMID- 10837658 TI - Transplantation of cells in matrices for tissue regeneration. AB - Tissue engineering is a field that has truly emerged in the last decade. It has brought together diverse technologies, e.g. cell culture, polymer chemistry and transplantation. The creation of matrices to guide tissue regeneration allows manipulation at several levels, i.e. the cells employed, the choice of polymer and the design of construct assembly methods. We present experience using such constructs to guide regeneration of diverse tissues, e.g. liver, intestine, urologic tissue, skin, cartilage, bone and cardiovascular structures. Emerging concepts in using cell/polymer constructs include the need for appropriate modeling of the micromechanical environments of different tissues, as well as the necessity of finding new strategies to achieve vascularization of tissues for transplant. Finally, the concept of applying tissue-engineered structures to non native sites is discussed. PMID- 10837659 TI - Pharmacodynamic aspects of sustained release preparations. AB - The sustained release (SR) mode of drug administration has certain features that have an important impact on the magnitude of the pharmacologic response: (a) it minimizes fluctuation in blood drug concentrations (i.e. between peak and trough). However, due to the pronounced non-linear relationship between drug concentration and pharmacologic effect (i.e. pharmacodynamics) the impact of this property differs considerably as a function of the shape of the pharmacodynamic profile and the position of the specific range of concentrations on the curve of this profile; (b) it produces a slow input rate which tends to minimize the body's counteraction to the drug's intervening effect on regulated physiological processes; and (c) it provides a continuous mode of drug administration. This important pharmacodynamic characteristic may produce, in certain cases, an opposite clinical effect than that attained by an intermittent (pulsatile) mode of administration of the same drug. For many drugs with non-concentration dependent pharmacodynamics, the exposure time, rather than the AUC, is the relevant parameter and it can therefore be optimized by SR preparations. The slow input function may minimize hysteresis in cases where the site of action is not in a rapid equilibrium with the blood circulation. The pharmacodynamics of the desired effect(s) and/or adverse effect(s) may also be influenced by the site of administration, especially in cases where the drug is delivered directly to its site of action. These factors demonstrate the important influence of the mode of administration on the pharmacological and clinical outcomes. In addition, they highlight the need to include these pharmacodynamic considerations in all stages from drug development to the optimization of their clinical use. PMID- 10837660 TI - Impact of pharmacodynamic variability on drug delivery(1). AB - Modern pharmaceutical delivery systems are intended to produce plasma drug concentration versus time profiles that result in optimum therapeutic efficacy and a minimum of drug concentration-dependent adverse effects. To accomplish this requires that the drug delivery rate and temporal profile be based on the pharmacokinetic and pharmacodynamic characteristics of the specific medicinal agent. Pharmacokinetic and pharmacodynamic parameters are subject to considerable interindividual variability. Whereas the importance of pharmacokinetic variability is generally recognized, the significance of pharmacodynamic variability (i.e., variability in the relationship between effect intensity and drug concentration) is not as widely appreciated. Pharmacodynamic variability is typically quite large, reproducible, and often substantially exceeds the relative magnitude of pharmacokinetic variability. This article consists of a review of how to assess pharmacodynamic variability, clinical examples of pharmacodynamic variability of drugs with a wide range of indications, and an outline of mechanisms of pharmacodynamic variability. PMID- 10837661 TI - Pharmacodynamics of variable patient compliance: implications for pharmaceutical value. AB - Pharmaceutical value, a multidimensional factor that includes proven efficacy, is now crucial to market success. A key parameter of value is reliability in use, determined by the several sources of variance in drug response, e.g. the Harter Peck model, based on linear pharmacometrics of theophylline, whose dose-response has an estimated 80% coefficient of variation, due mainly to variable pharmacokinetics and non-compliance. The main forms of non-compliance are multiday intervals between doses, the impact of which is difficult to assess because too little is known of the time-course of drug actions after dosing stops ('off-responses'). Omeprazole, the best-selling drug, is an exception, and its off-response data reveal markedly non-linear pharmacodynamics that appear to filter most of the variance that a linear model passes, projecting a big gain in reliability. The impact of variable compliance is attenuated by 'forgiveness', the post-dose duration of effective action (4-5 days for omeprazole) minus its recommended dosing interval. PMID- 10837662 TI - Role of dosage regimen in controlling indirect pharmacodynamic responses. AB - The role of drug delivery in controlling indirect pharmacodynamic responses was assessed via computer simulations and literature review. Simulations of responses related to basic indirect response mechanisms were performed for various drug input rates which allowed the importance of drug delivery rate on the overall pharmacodynamic response to be evaluated. Response versus time profiles of integrated or net responses and efficiency were examined. Rate of drug input has the greatest influence on the area under the effect curve when doses are larger and target drug concentrations are above the IC(50)/SC(50). The pharmacodynamics of drugs which elicit indirect pharmacologic responses such as corticosteroids, diuretics, growth hormone, erythropoietin and insulin indicate that sustained drug delivery enhances the therapeutic efficiency and pharmacodynamic availability. PMID- 10837663 TI - Pharmacodynamic aspects of drug administration. Tolerance development. AB - In the present investigations continuous intraduodenal infusion of levodopa has been given to patients with advanced Parkinson's disease during 6 months (seven patients) and 2.5 years (two patients). The plasma concentration and the effects on motility have been studied at the start, at 3 and 6 months and after 2.5 years. The individually titrated steady state plasma concentrations (C(ss)) for satisfactory mobility was found to be decreased by the time, and already after 1 month of intraduodenal infusion C(ss) was decreased by about 50%. The obtained data indicate an increased sensitivity with time to levodopa, interpreted as a consequence of disappearing tolerance by the controlled drug delivery. Blood pressure and heart rate were studied in spontaneous hypertensive rats during continuous administration of L-propranolol. A maximum reduction in heart rate of about 20% was seen, but no tolerance was observed. All animals showed, however, rebound effects that were at a maximum 3 days after drug withdrawal. Different PK/PD models have been fitted to the data and an indirect effect model seems to be the most appropriate. PMID- 10837664 TI - Pharmacodynamic aspects of peptide administration biological response modifiers. AB - Cytokines, growth factors and other recombinant proteins have been one of the most rapidly growing areas of pharmaceuticals. Further, the development of these bio-engineered drugs is occurring at an astonishing pace with rapid preclinical and clinical development and licensing by regulatory agencies. In addition, the availability of the gene sequences and rational drug design technologies have resulted in a rapid development of engineered genes, proteins and peptidomimetics. In contrast to traditional pharmacophores, which are developed based on the identification of the maximum tolerated dose (MTD), most recombinant proteins have abnormal biodistributions, and pharmacodynamic and pharmacokinetic attributes. Within this chapter, representative cytokines including interferon alpha (IFN-alpha), IFN-gamma and interleukin-2 are used to discuss the pharmacodynamic aspects of protein/peptide administration that are important in the development of these drugs. This includes the conceptual need for chronic immunoaugmentation for optimal therapeutic activity; the need to consider the pharmacokinetics of administration to optimize drug delivery and the nonlinear dose response relationship, which can result in a bell shaped dose response. Furthermore, these therapeutics have maximal potential in an adjuvant protocol and their development in combination with high-dose chemotherapy and stem cell rescue is discussed. The strategies for combination chemotherapy and immunotherapy, while holding great promise, require close attention to the pharmacodynamics of protein administration in order to impact on failure free and overall survival. PMID- 10837665 TI - An integrated pharmacokinetic-pharmacodynamic approach to optimization of R apomorphine delivery in Parkinson's disease. AB - R-apomorphine is a mixed dopamine D(1)/D(2) receptor agonist which is potentially useful in the management of Parkinson's disease. The delivery of R-apomorphine is complicated however by a number of pharmacokinetic and pharmacodynamic factors. This review describes the development of a transdermal iontophoretic delivery system for R-apomorphine on the basis of integrated pharmacokinetic pharmacodynamic (PK/PD) investigations in patients with idiopathic Parkinson's disease. The pharmacokinetics and metabolic pathways of R-apomorphine were determined following intravenous infusion of 30 ug kg(-1) in 15 min in 10 patients. A stepwise infusion protocol was used to determine the therapeutic window. A wide interindividual variability in both pharmacokinetics and pharmacodynamics and a narrow therapeutic concentration range were observed. This shows the need for individualized and carefully controlled delivery of R apomorphine in Parkinson's disease. Transdermal iontophoretic transport was studied both in vitro in human stratum corneum and dermatomed full skin and in vivo in patients with Parkinson's disease. These studies showed that the delivery of R-apomorphine by transdermal iontophoresis is feasible and furthermore that the rate of delivery can be carefully controlled by variation of the current density. It is concluded that the delivery of R-apomorphine by transdermal iontophoresis may be an attractive tool in future clinical pharmacological investigations in patients with Parkinson's disease aiming at characterization of the influence of chronic treatment and disease progression on the pharmacokinetics and pharmacodynamics. Ultimately these studies may result in a system which is suitable for clinical application. PMID- 10837666 TI - Future challenges for drug delivery research. PMID- 10837667 TI - Key issues in non-viral gene delivery. AB - The future of non-viral gene therapy depends on a detailed understanding of the barriers to delivery of polynucleotides. These include physicomechanical barriers, which limit the design of delivery devices, physicochemical barriers that influence self-assembly of colloidal particulate formulations, and biological barriers that compromise delivery of the DNA to its target site. It is important that realistic delivery strategies are adopted for early clinical trials in non-viral gene therapy. In the longer term, it should be possible to improve the efficiency of gene delivery by learning from the attributes which viruses have evolved; attributes that enable translocation of viral components across biological membranes. Assembly of stable, organized virus-like particles will require a higher level of control than current practice. Here, we summarize present knowledge of the biodistribution and cellular interactions of gene delivery systems and consider how improvements in gene delivery will be accomplished in the future. PMID- 10837668 TI - Application of membrane-active peptides for drug and gene delivery across cellular membranes. AB - Naturally occurring peptides and protein domains with amphipathic sequences play a dominant role in physiological, lipid membrane-reorganizing processes like fusion, disruption, or pore formation. More recently this capacity to modulate membrane integrity has been exploited for drug delivery into cells. Incorporation of synthetic membrane-active peptides into delivery systems has been found to enhance intracellular delivery of drugs including oligonucleotides, peptides, or plasmid DNA. In the majority of applications, the amphipathic peptides are designed to act after uptake by endocytosis, releasing the delivered agent from intracellular vesicles to the cytoplasm. Alternatively, peptides might mediate direct drug transfer across the plasma membrane. Although encouraging results have been obtained with the use of synthetic peptides to enhance cellular delivery of various compounds, the naturally evolved mechanisms observed in the entry of viruses or protein toxins are still far more efficient. For the development of improved synthetic peptides and carrier systems a better understanding of the molecular details of membrane-destabilization and reorganization will be essential. PMID- 10837669 TI - Lipid fusion in oligonucleotide and gene delivery with cationic lipids. AB - Delivery of oligonucleotides and genes to their intracellular targets is a prerequisite for their successful use in medical therapy. Cationic liposomes are among the most commonly used and promising delivery systems for oligonucleotides and genes. Lipid fusion plays an important role in the cationic liposome-mediated delivery of these compounds. Fusion is involved in the complex formation between the nucleotides and the lipids, in the interactions between extracellular materials with the complexes, as well as in the intracellular trafficking of the delivery system and its load. Since lipid fusion is such a crucial factor in polynucleotide delivery, its controlled use is important for the success in oligonucleotide and DNA delivery. In this article we are reviewing the current knowledge on lipid fusion phenomena associated with the delivery of oligonucleotides and genes. PMID- 10837670 TI - Nuclear import of polypeptides, polynucleotides and supramolecular complexes. AB - Nuclear import of macromolecules or particles is a strictly regulated phenomenon that is now understood in some depth at the molecular level. Transport across nuclear membranes is controlled by nuclear pores, with the involvement of cytoplasmic receptors and accessory molecules. Each nuclear pore complex is an assembly of multiple copies of at least 30 distinct proteins, with a total mass of approximately 10(7) kDa. Passive transport of small molecules through nuclear pores is unrestricted, but efficient uptake of macromolecules requires nuclear localization signals, which facilitate the interaction with cytoplasmic receptor proteins. The basic architecture of the nuclear pore and the mechanisms that regulate nuclear import are summarized, in relation to import of endogenous molecules and viruses. Subsequently, the significance of these mechanisms in controlling gene delivery is discussed. Access of DNA to the nucleus is a major barrier to the success of gene therapy, although viruses have evolved mechanisms to exploit the active transport machinery within the host cell. A key step in the future development of non-viral gene therapy will be the design and development of synthetic systems for active delivery of DNA to the nucleus. PMID- 10837671 TI - Mechanisms of enveloped virus entry into animal cells. AB - The ability of viruses to transfer macromolecules between cells makes them attractive starting points for the design of biological delivery vehicles. Virus based vectors and sub-viral systems are already finding biotechnological and medical applications for gene, peptide, vaccine and drug delivery. Progress has been made in understanding the cellular and molecular mechanisms underlying virus entry, particularly in identifying virus receptors. However, receptor binding is only a first step and we now have to understand how these molecules facilitate entry, how enveloped viruses fuse with cells or non-enveloped viruses penetrate the cell membrane, and what happens following penetration. Only through these detailed analyses will the full potential of viruses as vectors and delivery vehicles be realised. Here we discuss aspects of the entry mechanisms for several well-characterised viral systems. We do not attempt to provide a fully comprehensive review of virus entry but focus primarily on enveloped viruses. PMID- 10837672 TI - Extravasation of macromolecules. AB - Macromolecules can extravasate across the normal endothelium by transcapillary pinocytosis as well as by passage through interendothelial cell junctions, gaps or fenestrae. The main biological factors that control extravasation of a solute include regional differences in the capillary structures, the disease state of the organ or tissue, and the rate of blood and lymph supply. Physicochemical properties that are of profound significance in the extravasation of macromolecules are molecular size, shape, charge and hydrophilic/lipophilic balance (HLB) characteristics. Extravasation of small drugs, proteins, oligonucleotides and genes can be controlled by conjugating or forming complexes with macromolecular carriers. This requires a thorough understanding of the relationship between the chemical structures, physicochemical properties and the pharmacokinetics of both carrier and active molecules. This review article discusses the extravasation of macromolecules from the view points of pharmacokinetics and drug delivery systems, with the main emphasis on the extravasation across the liver, kidney and tumor capillaries. PMID- 10837673 TI - Control of tumour vascular permeability. AB - Several model tumour systems are now known to display increased vascular permeability compared with normal tissues, permitting their selective targeting using macromolecular drugs. Preliminary clinical observations suggest that this pathology may be reflected in at least some types of human cancer, and this may have important implications in facilitating macromolecular drug treatments, including antibody targeting and delivery of DNA for gene therapy. The enhanced permeability of tumour vasculature is thought to be regulated by tumour-secreted growth factors, with vascular permeability facor (VPF), also known as vascular endothelial growth factor (VEGF), emerging as a particularly likely candidate. VPF/VEGF is known to be an important regulator of tumour-angiogenesis in vivo, and it exerts its endothelium-specific effects via its receptors KDR/Flk-1 and Flt-1 on the endothelial cell membrane. Although the precise mechanism of VEGF's permeabilising action is not yet understood, it is likely to contribute to the enhanced permeability and retention (EPR) effect in tumours which is thought to underlie the anticancer activity of macromolecular drugs. PMID- 10837674 TI - Physiological considerations in the design of particulate dosage forms for oral vaccine delivery. AB - The gastrointestinal tract provides a variety of morphological (e.g. epithelial cells, mucus) and physiological (e.g. enzymes, pH, transporters) barriers to the absorption of peptides and proteins. Approaches to overcome these barriers have included the use of particulates which are taken up by specialized mechanisms present in M cells of the gastrointestinal tract. Due to its limited capacity, this approach has found particular application in the delivery of vaccines. In this review, morphological and physiological characteristics of the gastrointestinal tract which influence the design of particulates for oral delivery will be presented. Particulates have been designed to resist luminal factors responsible for limiting absorption and to target a specialized cell population, the M cells, within the gastrointestinal tract employing both physical and biological approaches (e.g. charge, size, hydrophobicity, surface ligands such as lectins). For vaccines, this approach may have 'particular' attraction due to the signal magnification which can be accomplished in the gut associated lymphoid tissue (GALT). Recent studies have demonstrated that epithelial cells can be converted to M cells following exposure to Peyer's patch lymphocytes. Future studies designed to identify the factor(s) responsible for transient conversion of epithelial cells to M cells could provide an approach to enhance efficiency of vaccine delivery. PMID- 10837675 TI - The effect of physical barriers and properties on the oral absorption of particulates. AB - This paper provides an in-depth review of the biological, immunological, and physicochemical barriers to the oral absorption of microparticles. The roles of the mucus and mucosal cell layers are discussed along with technologies and strategies that may be used to improve uptake, including attempts to determine the mechanisms of microparticulate uptake into enterocytes and membranous epithelial (M) cells. Considerations unique to the absorption of microparticles, such as particle size, surface characteristics, cell surface interactions, and the driving force for absorption of microparticles, are also addressed. Although the extent of absorption of microparticles is severely limited, this review provides insight into potential methods and technologies that have been and could be used to improve the oral absorption of microparticles. PMID- 10837676 TI - Polymeric carriers for oral uptake of microparticulates. AB - Delivery of vaccine antigens by the oral route is plagued with challenges. Much of the research has focused on the development of microparticles as antigen carriers to the gastrointestinal (GIT) mucosa. Polymers, either natural or synthetic, have been the class of compounds most often investigated for their ability to form microparticles containing antigen. A great deal of research has been performed using model microparticles composed of polystyrene. From this work it has become clear that microparticles are taken up and translocated across the GIT epithelium. Antigen carrying microparticles generated from both hydrophobic and hydrophilic polymers are able to induce significant immune responses after oral immunization. Although very little systematic work on the effects of the physicochemical properties of the polymer composing the microparticles has been done, enough is known to conclude that the surface of the polymeric microparticle can be decisive in determining the overall uptake of the microparticles. Charge and the hydrophobic/hydrophilic balance of the polymer are important physicochemical characteristics that determine the value of the polymer as a microparticulate carrier. This review examines the properties of polymeric matrices that make them viable candidates as oral vaccine delivery vehicles. PMID- 10837677 TI - Polymeric nano- and microparticles for the oral delivery of peptides and peptidomimetics. AB - Due to recent advances, numerous bioactive peptides are now available in large quantities. Administering these substances by the oral route appears as a formidable challenge due to their insufficient stability in the gastrointestinal tract and their poor absorption pattern. Several approaches have been investigated to improve their oral bioavailability. Among them, the use of polymeric particulate delivery systems (microparticles and nanoparticles) represents a promising concept. Encapsulating or incorporating peptides in particles should at least protect these substances against degradation and, in some cases, also enhance their absorption. The aim of this paper is to review the principal studies where peptide-loaded particles were administered by the oral route. The preparation methods and in vitro trials are presented and in vivo results are discussed with emphasis placed on the peptide blood levels reached or on the biological effects observed. Whether or not intact particles can be taken up and translocated to the systemic circulation is not the aim of this review. PMID- 10837678 TI - Specific and non-specific bioadhesive particulate systems for oral delivery to the gastrointestinal tract. AB - The oral route constitutes the preferred route for drug delivery. However, numerous drugs remain poorly available when administered by this route. In order to circumvent this problem, it has been proposed, successfully for several of them, to associate drugs to polymeric nanoparticulate systems (or small particles in the range of the micrometre in size) because of their propensity to interact with the mucosal surface. The present review focuses on the gastrointestinal bioadhesion of micro- and nanoparticles. Bioadhesion can be obtained by the building of either non-specific interactions with the mucosal surface, which are driven by the physicochemical properties of the particles and the surfaces, or specific interactions when a ligand attached to the particle is used for the recognition and attachment to a specific site at the mucosal surface. The relative merits of those systems are discussed. Their fate in the gastrointestinal tract, including at least three different pathways: (i) bioadhesion, (ii) translocation through the mucosa and (iii) transit and direct faecal elimination, is also presented. PMID- 10837679 TI - Evaluation of nano- and microparticle uptake by the gastrointestinal tract. AB - Numerous papers over the last two decades have demonstrated that particle uptake by the gastrointestinal tract is a reality. In addition, polymeric nano- and microparticles have proved to be useful delivery systems to enhance oral bioavailability of poorly absorbed drugs or to induce mucosal immune response. However, despite the amount of data available, no set criteria are available for the design of a good particulate carrier for oral delivery of peptides or antigens. This is partly due to the publication of conflicting and confusing data. The source of discrepancy is actually multiparametric (e.g., methodology, mode of evaluation, animal species) and is still not fully understood. The purpose of this review is to discuss the advantages and the limitations of the methodologies and the models used to evaluate gastrointestinal uptake of nano- and microparticles. PMID- 10837680 TI - Mucosal immunity of the gastrointestinal tract and oral tolerance. AB - To guard against disease, mucosal surfaces of the intestinal, respiratory and urogenital tracts are protected by a carefully regulated system of defenses known as the mucosal immune system. The hallmark of mucosal immunity is secretory IgA which can prevent infection and remove antigen crossing the mucosal barrier. IgE responses are also associated with mucosal immunity. In addition, a lymphocyte population resides within the mucosal epithelium, possibly forming a 'first line of defense' against infections. Oral administration of antigen can also induce antigen-specific peripheral immune tolerance, known as oral tolerance. Oral tolerance may prevent allergies, inflammation and other problems due to responses against food antigens or gut flora. It also has a tremendous potential in treating autoimmune and inflammatory diseases. The aim of this article is to present an overview of mucosal immunity. Specifically, the focus is on immunity along the gastrointestinal tract; however, similar principles are active in other mucosal tissues. PMID- 10837681 TI - Microparticle targeting to M cells. AB - M cells are epithelial cells specialized for sampling viruses and bacteria and perform a critical role in immune surveillance in mucosal tissues. The M cell apical surface is modified with less frequent microvilli and a thinner glycocalyx, and the basolateral membrane is invaginated to form an intraepithelial 'pocket' enclosing leukocytes. These alterations facilitate endocytosis and phagocytosis of microorganisms and transport across the epithelium. In certain tissues, M cell apical membranes exhibit distinct receptors which have been identified with monoclonal antibodies and lectins. Pathogenic bacteria and viruses can recognize the M cell surface and exploit its transcellular transport pathway to gain entry into host tissues. M cells sample and transport synthetic microspheres in a manner similar to microorganisms and thus play an essential role in the delivery of vaccines and drugs to mucosal lymphoid tissues. However, the efficiency of sampling orally delivered microparticles by M cells is low. Strategies are being developed to target microparticles to M cell apical surfaces using M cell-selective probes in order to increase sampling efficiency and bioavailability of drugs and vaccines. Such targeting strategies will likely be critical for the development of more effective vaccines and drugs administered by mucosal routes. PMID- 10837682 TI - The preparation and characterization of polymeric antigen delivery systems for oral administration. AB - Although polymeric delivery systems are well established for the oral administration of conventional drugs, they have not yet been commercially developed for vaccine delivery. The problems inherent with the oral route of delivery, including low pH, gastric enzymes, rapid transit and poor absorption of large molecules, has made the goal of oral delivery of antigens very challenging. Nevertheless, several polymeric delivery systems for the oral administration of vaccines are currently being evaluated, including microencapsulation in poly(lactide-co-glycolides), alginates, polyanhydrides, starch, polymethacrylates, polyamino acids and enteric coating polymers. These approaches are designed to protect the antigen in the gut, to target the antigen to the gut associated lymphoid tissue, or to increase the residence time of the antigen in the gut through bioadhesion. Each of these approaches is discussed in relation to antigen encapsulation and integrity, process reproducibility, ease of preparation and encapsulation efficiency. Potential problems associated with the scale-up of these approaches are also briefly addressed. Of particular relevance are the prospects for the application of these formulation processes for commercial development. PMID- 10837683 TI - Microparticles and polymers for the mucosal delivery of vaccines. AB - Because microparticles are taken up across the gastrointestinal tract following oral administration, they may be exploited for the oral delivery of labile compounds. For example, microparticles have been used for the oral delivery of peptide and protein drugs, and have resulted in improvements in bioavailability. In addition, microparticles have also been exploited for the oral delivery of vaccines, inducing potent immune responses and protective immunity. However, the extent of uptake of microparticles across the gut may limit their potential for oral delivery. Therefore, intranasal immunization is an attractive approach for the induction of mucosal immunity. Microparticles have also been used for the delivery of vaccines to the respiratory tract. In addition to the use of small microparticles to target antigens to mucosal lymphoid tissues, polymeric coatings may also be employed simply to protect antigens against degradation during transit in the gut. Hence, oral delivery using polymers is not necessarily dependent on the uptake of the delivery system across the gut. Recent studies have indicated that a number of polymeric delivery systems possess significant potential for the development of mucosally administered vaccines. However, further work is needed in a number of areas, including the stabilization of antigens within the polymeric delivery systems. PMID- 10837684 TI - Uptake and adjuvant activity of orally delivered saponin and ISCOM vaccines. AB - Saponins are a highly heterogenous group of glycosides which are common in plants and have been known to have adjuvant properties since the 1920s. The immunostimulating complex or ISCOM is a particulate adjuvant/antigen delivery system. ISCOMs are open cage-like complexes typically with a diameter of about 40 nm which are built up by cholesterol, lipid, immunogen and saponins from the bark of Quillaia saponaria Molina (soap bark tree). ISCOMs and saponins are used as adjuvants in some commercial veterinary vaccines and have been examined as adjuvants in a large number of human experimental vaccines. This review describes the current status and potential of saponin and ISCOMs as adjuvants for orally administered vaccines with special reference to the induction of local and systemic immune responses and interactions with the intestinal epithelium. The structure and composition of saponins and ISCOMs will also be reviewed. PMID- 10837685 TI - Oral particulate delivery: status and future trends. AB - Oral delivery of complex molecules such as peptides and proteins is one of the most intensively studied research subjects. However, its success has been hampered by the degradation of these molecules in the gastrointestinal tract. As a result, various types of particulate systems such as biodegradable microspheres and liposomes have been proposed as potential delivery vehicles to protect these drugs in the gastrointestinal tract. Unfortunately, these particulates generally display low oral absorption efficiencies. This renders most encapsulated drugs ineffective after oral administration and therefore remains as the main obstacle to their practical application as oral delivery vehicles. In attempts to improve particle absorption efficiency, strategies such as using mucoadhesive polymers and targeted delivery systems have been studied in animals. In this chapter, the usefulness of these strategies are briefly reviewed. Potential applications and future prospects of particulates in oral delivery of complex molecules are also highlighted. PMID- 10837686 TI - Electropermeabilization of cell membranes. AB - A position dependent modulation of the membrane potential difference is induced when an electric field is applied to a cell. When cells are submitted to short lived electric field pulses with an overcritical intensity, a local membrane alteration is induced, which may reseal. Its molecular definition remains unknown. A free exchange of hydrophilic molecules takes place across the membrane. A leakage of cytosolic metabolites is present. However, a loading of polar drugs into the cytoplasm is obtained. A short description of the processes affecting the cell membrane organization is given. Lipids appear as the primary target of the field effect as in the case of liposomes. Nevertheless membrane proteins appear to be affected by a direct or by a back effect. The permeabilized state is long lived. The cell metabolism plays indeed a critical role in the recovery. The cell viability can be nevertheless preserved. PMID- 10837687 TI - Theory of electrical creation of aqueous pathways across skin transport barriers. AB - Experimental studies have shown that application of electrical pulses to human skin that result in U(skin)>30 V for durations of about 1 ms or longer causes a large decrease in electrical resistance within microseconds, followed in seconds by an increase in molecular transport of water-soluble molecules. Local transport regions (LTRs), within which molecular transport is concentrated, mostly form away from the skin's appendages and rete pegs. Theoretical attempts to explain this behavior involve electrically created aqueous pathways ("pores"). For short (about 1 ms) "high voltage" (HV) pulses leading to about U(skin)>50 V, it was hypothesized that such pulses cause electroporation of the multilamellar lipid bilayer membranes of the skin's stratum corneum (SC). Much of the present experimental evidence supports the more specific hypothesis that such pulses create "straight through aqueous pathways", mostly within LTRs, that perforate the SC lipid bilayers and pass through the interiors of hydrated corneocytes. Theoretical estimates of the localized heating within LTRs predict relatively small temperature rises. The theory of LTR formation is incomplete, with both stochastic and deterministic models under consideration. Moderate voltage (MV) pulses leading to about 5400 Da) hydrophobic drugs in the brain, by performing active back-transport of these drugs to the blood. Extensive experiments with in vitro models and with knockout mice lacking blood-brain barrier P-glycoprotein or other animal models treated with blockers of P glycoprotein have fully confirmed this hypothesis. Absence of functional P glycoprotein in the blood-brain barrier leads to highly increased brain penetration of a number of important drugs. Depending on the pharmacological target of these drugs in the central nervous system (CNS), this can result in dramatically increased neurotoxicity, or fundamentally altered pharmacological effects of the drug. Given the variety of drugs affected by P-glycoprotein transport, it may be of tremendous therapeutic value to apply these insights to the development of drugs that should have either very poor or very good brain penetration, whichever is preferred for pharmacotherapeutic purposes. The clinical application of P-glycoprotein blockers should also be considered in order to improve the blood-brain barrier permeability of certain drugs that currently display insufficient brain penetration for effective therapy. PMID- 10837716 TI - The blood-brain barrier efflux transporters as a detoxifying system for the brain. AB - The role played by efflux transport systems across the blood-brain barrier (BBB) in the disposition of xenobiotics in the brain is described. Several drugs and organic anions are transported across the BBB via P-glycoprotein and other carrier-mediated efflux transport systems. Studies using in vitro cultured brain capillary endothelial cells, kinetic analysis, and mdr1a gene knock-out mice have shown that P-glycoprotein, located on the BBB, restricts the entry of vincristine and quinidine to the brain. Brain microdialysis studies have demonstrated that the brain interstitial fluid (ISF) concentrations of quinolone antibiotics are significantly lower than their corresponding unbound serum concentrations. A distributed model analysis supports the finding that efflux transport systems on the BBB restrict distribution of 3'-azido-3'-deoxythymidine (AZT), 2',3' dideoxyinosine (DDI), and quinolone antibiotics. A brain efflux index (BEI) method has been developed to provide direct evidence of an efflux transport system for carrying substrates from the cerebrum to the circulating blood across the BBB. The BEI method revealed the existence of carrier-mediated efflux organic anion transport systems for compounds such as p-aminohippuric acid, AZT, DDI, taurocholic acid, BQ-123, and estron sulfate. Moreover, cerebral neurotransmitters such as gamma-aminobutyric acid, L-glutamic acid, and L aspartic acid are transported from brain to the circulating blood in the intact form via a carrier-mediated efflux transport system. The BBB not only restricts nonspecific permeation from the circulating blood to the brain, but also functions as an active efflux transport system for xenobiotics. Accordingly, the BBB plays a very important role by pumping xenobiotics and some endogenous compounds out of the brain, acting as a central nervous system (CNS)-specific detoxifying system supporting and maintaining normal cerebral function. PMID- 10837717 TI - Microdialysis for pharmacokinetic analysis of drug transport to the brain. AB - The intracerebral microdialysis technique represents an important tool for monitoring free drug concentrations in brain extracellular fluid (brain(EcF)) as a function of time. With knowledge of associated free plasma concentrations, it provides information on blood-brain barrier (BBB) drug transport. However, as the implantation of the microdialysis probe evokes tissue reactions, it should be established if the BBB characteristics are maintained under particular microdialysis experimental conditions. Several studies have been performed to evaluate the use of intracerebral microdialysis as a technique to measure drug transport across the BBB and to measure regional pharmacokinetics of drugs in the brain. Under carefully controlled conditions, the intracerebral microdialysis data did reflect passive BBB transport under normal conditions, as well as changes induced by hyperosmolar opening or by the presence of a tumor in the brain. Studies on active BBB transport by the mdr1a-encoded P-glycoprotein (Pgp) were performed, comparing mdr1a(-/-) with wild-type mice. Microdialysis surgery and experimental procedures did not affect Pgp functionality, but the latter did influence in vivo concentration recovery, which was in line with theoretical predictions. It is concluded that intracerebral microdialysis provides meaningful data on drug transport to the brain, only if appropriate methods are applied to determine in vivo concentration recovery. PMID- 10837718 TI - Recent advances in the brain targeting of neuropharmaceuticals by chemical delivery systems. AB - Brain-targeted chemical delivery systems represent a general and systematic method that can provide localized and sustained release for a variety of therapeutic agents including neuropeptides. By using a sequential metabolism approach, they exploit the specific trafficking properties of the blood-brain barrier and provide site-specific or site-enhanced delivery. After a brief description of the design principles, the present article reviews a number of specific delivery examples (zidovudine, ganciclovir, lomustine benzylpenicillin, estradiol, enkephalin, TRH, kyotorphin), together with representative synthetic routes, physicochemical properties, metabolic pathways, and pharmacological data. A reevaluated correlation for more than 60 drugs between previously published in vivo cerebrovascular permeability data and octanol/water partition coefficients is also included since it may be useful in characterizing the properties of the blood-brain barrier, including active transport by P-glycoprotein. PMID- 10837719 TI - A novel chemical delivery system for brain targeting. AB - Two different chemical approaches for brain drug delivery and targeting are proposed in the present review. One is a chemical drug delivery using a ring closure reaction to the hydrophilic quaternary thiazolium compound in the brain. The other is a chemical drug targeting utilizing the nutrient receptor (transporter) system on the blood-brain barrier. The brain delivery system has been optimized and it was demonstrated that the brain delivery of three drugs, a drug for Parkinson's disease, an excitatory amino acid antagonist and a free radical scavenger, were improved by the conjugation with cis-2 formylaminoethenylthio derivatives in vivo. As for the brain targeting system, it was demonstrated that the conjugation with L-Glu improved the drug's brain distribution via the L-Glu excitatory and/or transport receptors in vitro and in vivo. These findings suggest that the concepts of two chemical approaches will contribute to the development of new central nervous system drugs. PMID- 10837720 TI - Carrier-mediated or specialized transport of drugs across the blood-brain barrier. AB - In the drug development process, it remains a difficult task to regulate the entry of the drugs. However, recent progress in studies of the transporter mediated influx and efflux of endogenous and exogenous compounds, including synthetic drugs, across the blood-brain barrier (BBB) is beginning to provide a rational basis for controlling drug distribution to the brain. This paper describes mechanisms established in the last decade for carrier-mediated influx and efflux of drugs and endocytosis of biologically active peptides across the BBB. The transport systems at the BBB described here are the uptake transporters for nutrients, such as amino acids and hexoses, monocarboxylates, amines, carnitine and glutathione and efflux transporters, such as P-glycoprotein and multiple organic anion transporters. Delivery of cationized peptides across the BBB via adsorptive-mediated endocytosis is also described. By utilizing such highly specific transport mechanisms, it should be possible to establish strategies to regulate the entry of candidate drugs, including peptides, into the brain. PMID- 10837721 TI - Penetration of neurotrophins and cytokines across the blood-brain/blood-spinal cord barrier. AB - Now that peptides are no longer considered too large to cross the blood-brain barrier, attention has turned to the possibility that larger substances like polypeptides might also enter the central nervous system (CNS). This review summarizes evidence showing that many cytokines and neurotrophins not only enter the brain but also enter the spinal cord, sometimes faster than into the brain. PMID- 10837722 TI - Vector-mediated drug delivery to the brain. AB - Vector-mediated drug delivery to the brain employs the chimeric peptide technology, wherein a non-transportable drug is conjugated to a blood-brain barrier (BBB) transport vector. The latter is a modified protein or receptor specific monoclonal antibody that undergoes receptor-mediated transcytosis through the BBB in vivo. Conjugation of drug to transport vector is facilitated with either chemical linkers, avidin-biotin technology, polyethylene glycol linkers, or liposomes. Multiple classes of therapeutics have been delivered to the brain with the chimeric peptide technology, including peptide-based pharmaceuticals, such as a vasoactive intestinal peptide analog or neurotrophins such as brain-derived neurotrophic factor, antisense therapeutics including peptide nucleic acids, and small molecules incorporated within liposomes. The successful delivery of a drug through the BBB in vivo requires special molecular formulation of the drug. Therefore, it is important to merge central nervous system drug discovery and delivery as early as possible in the overall CNS drug development process. PMID- 10837723 TI - Delivery of diagnostic agents: achievements and challenges. PMID- 10837724 TI - Delivery of gamma-imaging agents by liposomes. AB - Liposomes are spherical bilayers which spontaneously form when water is added to a dried lipid mixture. Much progress has been made in the use of liposomes as vehicles for the delivery of gamma imaging agents. These radiolabeled liposomes have a wide variety of potential diagnostic uses including the detection of sites of infection, inflammation, gastrointestinal bleeding, tumor, cardiac blood pool imaging and lymphoscintigraphy. The ability to modify the surface of the liposomes permits the customization of liposome formulations for each particular diagnostic use. PMID- 10837725 TI - Delivery of contrast agents for magnetic resonance imaging, computed tomography, nuclear medicine and ultrasound. AB - This review describes and provides an overview of, and recent developments in, the area of liposomal contrast agents as applied to the diagnostic specialities of magnetic resonance, computed tomography, nuclear medicine and ultrasonography. Following a brief background of the clinical context and role of contrast agents in these various modalities, the review presents, where appropriate, a discussion of the theoretical background, as well as the critical design considerations pertinent to the rational and optimal delivery of contrast using lipid vesicles. Each section describes recent applications not covered by earlier reviews and concludes with a brief discussion of questions that remain unanswered or unresolved, that would merit further study. PMID- 10837726 TI - Delivery of contrast agents for positron emission tomography imaging by liposomes. AB - Liposomes encapuslating positron emitters are applicable for diagnostic imaging and are useful to investigate the real-time liposomal trafficking in vivo. Long circulating liposomes encapsulaing [2-(18)F]-2-fluoro-2-deoxyglucose were administrated to tumor-bearing mice, and a PET scan was performed. Small-sized long-circulating liposomes (100 nm) tended to accumulate in tumor tissues of tumor-bearing mice as compared with conventional liposomes. Then the size effect on trafficking of long-circulating liposomes was investigated. Large-sized liposomes (>300 nm) accumulated in liver and spleen in a time dependent manner. On the contrary, small-sized ones (<200 nm) were transiently accumulated in the liver right after injection, but the accumulation decreased time dependently, suggesting that, although the majority of small long-circulating liposomes remain in bloodstream, some extravasate once into interstitial spaces in liver which re enter into bloodstream again. Next the trafficking of so-called long-circulating liposomes, i.e., liposomes modified with ganglioside GM1, palmityl glucuronide (PGlcUA), and polyethylene glycol (PEG), in tumor-bearing mice was examined. The accumulation of all three kinds of long-circulating liposomes in liver decreased time-dependently, and PGlcUA-liposomes could avoid liver-trapping the most efficiently. Tumor accumulation of liposomes was obvious for PGlcUA-liposomes and PEG-liposomes from immediately after injection, but not for GM1-liposomes. Finally, the trafficking of differently charged liposomes was investigated in normal mice. The accumulation of positively charged liposomes containing 1,2 dimyristyloxypropyl-3-dimethyl-hydroxyethyl bromide was different from that of neutral and negatively charged DCP-liposomes. The agglutinability of and serum protein ginding to positively charged liposomes were marked, suggesting that these factors affect the high accumulation of DMRIE-liposomes in liver. Non invasive PET analysis of liposomal trafficking is beneficial for obtaining information about liposomal drug delivery, and long-circulating liposomes might be useful for diagnostic tumor imaging by PET. PMID- 10837727 TI - Antibodies as delivery systems for diagnostic functions. AB - Antibodies are highly specific targeting agents. Therefore, they are invaluable for in vitro and in vivo diagnostic applications. With the advent of monoclonal antibody technology, the utilization of antibodies has increased dramatically in almost every field of biological sciences. The present review describes the utility of monoclonal antibodies primarily in the cardiovascular diseases. Monoclonal antimyosin antibodies have been developed for noninvasive scintigraphic imaging of equivocal acute myocardial infarction. They have been negative charge modified to provide quicker in vivo visualization of the targeted antibody, as well as being applied for diagnosis of other cardiomyopathies that have disruption of myocardial cell membrane as an obligatory component of the disease. The radiolabeling techniques developed initially for myocardial necrosis imaging have also been applied for imaging of intravascular blood clots and atherosclerotic lesions. The applications of antimyosin, antifibrin and anti atherosclerotic lesion specific monoclonal antibodies have all achieved initial clinical verification of their efficacy to target the respective lesions. However, to date, only antimyosin has been approved by the FDA for commercialization. Others must await additional studies to unequivocally verify the clinical utilities. PMID- 10837728 TI - Polymeric micelles as carriers of diagnostic agents. AB - This review deals with diagnostic applications of polymeric micelles composed of amphiphilic block-copolymers. In aqueous solutions these polymers spontaneously form particles with diameter 20-100 nm. A variety of diagnostic moieties can be incorporated covalently or non-covalently into the particulates with high loads. Resulting particles can be used as particulate agents for diagnostic imaging using three major imaging modalities: gamma-scintigraphy, magnetic resonance imaging and computed tomography. The use of polyethyleneoxide-diacyllipid micelles loaded with chelated (111)In/Gd(3+) as well as iodine-containing amphiphilic copolymer in percutaneous lymphography and blood pool/liver imaging are discussed as specific examples. PMID- 10837729 TI - Avidin-biotin system for delivery of diagnostic agents. AB - Radiolabeled monoclonal antibodies specific for tumor-associated antigens are used for the diagnosis and therapy of malignant tumors. However, the blood clearance of antibodies is slow and tumor-to-nontumor ratios of radioactivity in the current system are not high. To increase tumor-to-nontumor ratios, the concept of pretargeting has been proposed. The strategy is based on the separate administration of the monoclonal antibody and the radiolabel. Because avidin and streptavidin bind to biotin selectively with extremely high affinity, the avidin biotin system has been applied for pretargeting. With this method, high tumor-to nontumor ratios are reported, not only in animal experiments, but also in clinical trials. The avidin-biotin system is also used to reduce the background radioactivity of the directly labeled antibody as well as radioimmunoguided surgery. PMID- 10837730 TI - Delivery of diagnostic agents for gamma-imaging. AB - The administration of radiolabeled compounds with gamma-emitting radionuclides allows real time pursuit of in vivo behavior of the compounds by external counting of the radioactivity. If the compounds possess a specific localization mechanism to target tissue, the image reflects the specific function of the target. This technique has widely been applied in diagnostic nuclear medicine combined with appropriate radionuclides for clinical use. Two strategies have been developed to design radiolabeled compounds for targeted imaging. One constitutes tethering radiolabeled compounds to carrier molecules that possess specific localization mechanisms to target tissue. Another approach constitutes chemical design of radiolabeled compounds, especially metallic compounds, that possess specific localization characteristics to the target per se. In this manuscript, progress of chemical design of radiolabeled compounds for targeted imaging is briefly reviewed. Emphasis is laid on chemical design of radiolabeled antibodies for targeted imaging. PMID- 10837731 TI - Delivery of diagnostic agents for magnetic resonance imaging. AB - A review of contrast agents used for magnetic resonance imaging was made with regard to methods of drug delivery using published literature. Since the clinical approval of Gd-DTPA in 1988, there has been extensive research towards developing organ- and tissue-specific contrast agents. Targeting strategies have consistently improved along with improvements in nuclear medicine imaging, and a broad spectrum of potential agents has accumulated. Liver, blood-pool targeted, and, due to their inherent convenience of delivery, intraorally administered gastrointestinal agents have been developed or are being developed. For intravenous contrast agents, collective magnetic labels with modifications for some specificities results in the larger-sized agents which can be an obstacle for the agent in accessing the targeted cells. In conclusion, the next step in the development of specific contrast agents for clinical use is to improve non specific delivery to the extra-capillary space adjacent to targeted cells. PMID- 10837732 TI - Targeted delivery of gas-filled microspheres, contrast agents for ultrasound imaging. AB - Gas-filled microbubbles, with the size of several micrometres, are strong scatterers of ultrasound waves used in diagnostic imaging. Application of these microbubbles as ultrasound contrast materials is discussed, in view of the design of materials capable of selectively targeting the diseased tissues/organs. Methods of preparation, mechanisms of action, biodistribution and stability in vitro and in vivo are reviewed. Targeted microbubbles with various ligands (antibodies, peptides) attached to the shells have been prepared and tested in vitro and in vivo. Examples of specific application in diagnostic imaging and possible therapeutic use are discussed. PMID- 10837733 TI - Delivery of diagnostic agents in computed tomography. AB - Computed tomography is a reliable, widely available imaging modality with high spatial resolution. Except for trauma detection, in practically all cases contrast agents are administered for improving image quality. The presently available agents are nonspecific compounds with distribution in the extracellular space. Specificity (i.e. high contrast in selected tissues) is only possible when the pharmacokinetic behavior (peak levels at the region of the interest) is followed on a time scale which counts in seconds. If the peak time is missed, a second injection of contrast material is necessary. The objective of extensive research efforts have therefore been the search for tissue-specific contrast agents which should result both in higher and in longer lasting local concentrations. For the tissue targets, liver, spleen, lymph nodes and the blood pool, efficient contrast enhancing compounds have been described utilizing different active transport systems or passive diffusion-limiting molecular sizes. The agents include water-soluble metal chelates, iodinated polymeric compounds and contrast agent-carrying liposomes. The remaining task for the future will be to improve the tolerability of the contrast agents to such an extent that the side-effect incidence and severity is low enough to allow for the use of these agents in patients. PMID- 10837734 TI - Delivery of radioligands for positron emission tomography (PET) in the central nervous system. AB - Positron emission tomography (PET) is an imaging technique to monitor the delivery of tracers labeled with positron emitters ((11)C, (13)N, (15)O and (18)F). A wide variety of probes have been labeled to measure biochemical and physiological parameters in the central nervous system (CNS), such as glucose and oxygen metabolism, protein synthesis, blood flow, and neurotransmitter receptor functions. The delivery of labeled compounds to the target tissue, which directly reflect the distribution and kinetics patterns, especially to the neurotransmitter receptors is modulated by several factors, such as regional cerebral blood flow (rCBF), peripheral metabolism, and neurotransmitter concentration in the synaptic cleft. These factors provide misunderstanding of the apparent results, which do not reflect the true state of the CNS. The present paper will summarize several factors that affect the delivery of labeled compounds related to the neurotransmitter receptors in the CNS. PMID- 10837735 TI - Utilization of metabolic, transport and receptor-mediated processes to deliver agents for cancer diagnosis. AB - The use of radiopharmaceuticals for the non-invasive diagnosis of cancer has been established in diagnostic radiology over the last few decades. In particular, with the use of sophisticated imaging modalities such as PET and SPECT and a myriad of radioisotopes, advances have been made in the detection and treatment of cancer. This article focuses on three available methods of tumor targeting with radiopharmaceuticals: the utilization of metabolic, transport and receptor mediated processes to deliver agents for cancer diagnosis. With selected reference to both clinically approved drugs and drugs currently under development, methods of uptake are presented either in terms of flow, metabolic or receptor mediated uptakes. A section of this article is devoted to the monitoring of cancer therapy regimes using radiopharmaceuticals. This review also discusses some mechanistic approaches available in radiopharmaceutical chemistry to be able to effectively diagnose and treat sufferers of cancer in the future. PMID- 10837736 TI - Radionuclide imaging of cardiac pathology: a mechanistic perspective. AB - Cardiac imaging is performed for a variety of indications using different radiotracers and ligands. The most common is the use of thallium-201 and Tc labeled tracers for myocardial perfusion imaging. Its uses are manifold, and range from the diagnosis of coronary artery disease to the risk stratification of patients and prognostication. Ischemia is being imaged with nitroimidazole compounds, myocardial necrosis with antimyosin and myocardial infiltration with gallium-labeled leukocytes and somatostatin receptor analogs. The status of myocardial innervation has been evaluated with radiolabeled MIBG. Metabolism in the heart in the normal and ischemic state can be studied with PET and SPECT using fatty acid and glucose analogs. The expression of accessory molecules on the surface of cardiac myocytes in patients with cardiac allograft rejection is now possible. Clearly, much has been accomplished for imaging the heart and pathology related to it. The past has been satisfying, the present interesting and the future promises to be even more exciting! PMID- 10837737 TI - Liposomes for scintigraphic detection of infection and inflammation. AB - Liposomes are small vesicles consisting of one or more concentric lipid bilayers enclosing discrete aqueous spaces. Liposomes potentially serve as carriers for radiolabels to visualize pathological processes scintigraphically. Research performed during the last two decades has revealed insight on the characteristics of liposomes with regard to their tissue distribution after i.v. injection. Various approaches have been developed to radiolabel liposomes with gamma emitting radionuclides such as Ga-67, In-111 and Tc-99m. Preferably, liposomes are labeled according to the 'after-loading' method. The radionuclide can either be encapsulated in the aqueous interior or coupled to the lipid bilayer. The potential of radiolabeled liposomes to image infectious and inflammatory foci has been demonstrated in a variety of preclinical studies. Recently, a study in patients suspected of having infectious or inflammatory disease has been initiated. Preliminary clinically results exemplify the potential of Tc-99m-PEG liposomes to image infection and inflammation. PMID- 10837738 TI - Targeted imaging of infection. AB - The prompt identification, localization and characterization of focal sites of infection in the patients with fever is a critical step in clinical management, particularly, when localizing symptoms are not present. Although the classic signs of inflammation are suitable to localize injury at superficial sites or in the extremities, inflammation of internal structures, such as in the brain, chest and abdomen, can be difficult to localize without additional diagnostic procedures. Tissue injury, regardless of cause or anatomical site, results in a complex series of physiologic changes that we recognize as the inflammatory response. The inflammatory response is characterized by a series of biochemical events in the insulted cells and surrounding structures that results in the three major pathophysiological components: increased tissue perfusion, increased vascular permeability, and leukocytic exudation. Exploitation of the early attributes of the inflammatory process are not sufficient for the routine detection of inflammation. Currently, reagents for targeting infection represent cellular or protein components involved in the inflammatory process. Such approaches have met with some success as these agents comprise integral parts of the complex phenomena known as inflammation. This same fact also limits their utility. Improved agents for targeting infection will likely be based on small molecules whose diffusion into the lesion is not hindered by molecular size constraints and which bind to molecular targets at the site of infection/inflammation. In general, the lower molecular weight should also lead to enhanced blood clearance, avoiding elevated blood pool activity which contributes to background. New agents should also obviate the need to handle blood, as this represents potential hazards to both patient and the medical personnel alike. PMID- 10837739 TI - Delivery of imaging agents into brain. AB - Delivery of diagnostic agents to the central nervous system (CNS) poses several challenges as a result of the special features of CNS blood vessels and tissue fluids. Diffusion barriers exist between blood and neural tissue, in the endothelium of parenchymal vessels (blood-brain barrier, BBB), and in the epithelia of the choroid plexuses and arachnoid membrane (blood-CSF barriers), which severely restrict penetration of several diagnostic imaging agents. The anatomy of large vessels can be imaged using bolus injection of X-ray contrast agents to identify sites of malformation or occlusion, and blood flow measured using MRI and CT, while new techniques permit analysis of capillary perfusion and blood volume. Absolute quantities can be derived, although relative measures in different CNS regions may be as useful in diagnosis. Local blood flow, blood volume, and their ratio (mean transit time) can be measured with high speed tomographic imaging using MRI and CT. Intravascular contrast agents for MRI are based on high magnetic susceptibility agents such as gadolinium, dysprosium and iron. Steady-state imaging using agents that cross the BBB including (123)I- and (99m)Tc-labelled lipophilic agents with SPECT, gives a 'snapshot' of perfusion at the time of injection. Cerebral perfusion can also be measured with PET, using H(2)(15)O, (11)C- or (15)O-butanol, and (18)F-fluoromethane, and cerebral blood volume measured with C(15)O. Recent advances in MRI permit the non-invasive 'labelling' of endogenous water protons in flowing blood, with subsequent detection as a measure of blood flow. Imaging the BBB most commonly involves detecting disruptions of the barrier, allowing contrast agents to leak out of the vascular system. Gd-DTPA is useful in imaging leaky vessels as in some cerebral tumors, while the shortening of T(1) by MR contrast agents can be used to detect more subtle changes in BBB permeability to water as in cerebral ischemia. Techniques for imaging the dynamic activity of the brain parenchyma mainly involve PET, using a variety of radiopharmaceuticals to image glucose transport and metabolism, neurotransmitter binding and uptake, protein synthesis and DNA dynamics. PET methods permit detailed analysis of regional function by comparing resting and task-related images, important in improving understanding of both normal and pathological brain function. PMID- 10837740 TI - Approaches and agents for imaging the vascular system. AB - Several classes of vascular imaging agents are described: (1) liposome-based blood cell mimetics; (2) plasma protein mimetics; (3) small molecules that bind to plasma proteins in the circulation. The characteristic features of the different agents are described and critically compared, including the advantages and potential pitfalls of each individual type. PMID- 10837741 TI - Subcutaneous and intravenous delivery of diagnostic agents to the lymphatic system: applications in lymphoscintigraphy and indirect lymphography. AB - Lymph node status is important in the staging of many malignancies. Although tissue characterization by histologic analysis of biopsy samples may improve staging, noninvasive staging is more acceptable to both patients and clinicians. Several imaging techniques may serve this goal. Modern noninvasive techniques such as computed tomography and magnetic resonance detect lymph node abnormality by nodal enlargement, but that does not always imply malignant involvement. On the other hand, many nodes are infiltrated or replaced by tumour without change in size. This becomes a serious diagnostic defect by these modalities. Consequently, attention has been focused to develop contrast agents and radiolabelled complexes for better cancer detection as well as characterization of individual tumours in lymph nodes. For delivery of such materials to regional lymph nodes one can take advantage from the distinct physiological function of the lymphatic capillaries. The thin-walled and fenestrated lymphatic microvessel is easily penetrated by particulate and macromolecular agents after injection into the extracellular space. Once inside the vessel, materials that are transported with the lymph either specifically target certain nodal elements (e.g. neoplastic cells) or become cleared by macrophages located in the lymph nodes. Indeed, interstitial delivery of diagnostic agents have been of benefit in determining regional spread of cancer and assessing lymphatic function either by lymphoscintigraphy or indirect lymphography. On the other hand, development of contrast materials that can reach lymph nodes after a single intravenous injection is highly desirable because of the large number of lymph nodes in the body and access being difficult to most of them. Today, a number of contrast agents exist that can reach a vast array of lymph nodes in the body, particularly those that are not readily accessible for histologic evaluation, after a single intravascular injection to help distinguish between normal and tumour-bearing nodes or reactive and metastatic nodes with magnetic resonance. In this article we critically examine advantages and limitations of both subcutaneous and intravenous routes of injection for the delivery of diagnostic agents to the lymphatic system. PMID- 10837742 TI - Preface. PMID- 10837743 TI - Comparative placental structure. AB - The primary function of all placentas is to act as an interface between the mother and fetus that allows, and even promotes, appropriate metabolic exchanges. This function is accomplished by bringing maternal and fetal blood into close apposition while maintaining separation of the maternal and fetal circulatory systems. Despite the common physiological functions shared by placentas, however, examination of placental morphology from different animal groups reveals a remarkable diversity of species-specific structural organization.The separation of fetal and maternal blood is always maintained by an elaboration of extraembryonic fetal tissues that cover fetal blood vessels. In some species the outermost layer of this fetal tissue, the trophoblast, is in direct contact with maternal blood. In many other species uterine tissues also contribute to the selective barrier separating the two blood systems. In addition to morphological variation among placentas of different animal groups, placentas undergo substantial structural modifications during pregnancy in a single species. In some animals different types of placentas function successively, or concurrently during a single pregnancy.As a result of these myriad details of placental structure, effective evaluation of fetal-maternal transfer of drugs must consider not only the components of the interhemal barrier of the fully developed placenta characteristic for each species, but also the placental structures functioning at each gestational stage of the fetus. PMID- 10837744 TI - Techniques to study human placental transport. AB - Pregnancy in the 20th century involves women of many age groups from early teens to the fourth or fifth decade. Modern medicine and in vitro fertilization techniques have increased options for pregnancy and childbirth. Pregnancy is a dynamic state, and medical concerns may involve disorders of the fetus and mother requiring medications and special nutrients. Therefore, different techniques have been developed to evaluate the placental transfer of drugs and nutrients using tissues and cells derived from human placenta. These include (a) isolated tissues and cells to study placental transport, (b) primary and malignant trophoblast cell cultures and (c) biophysical methods for studying placental transport. Also, convenient study models have been developed to evaluate placental transfer of safe drugs in pregnant women. Some of the drugs studied by these techniques and models include (a) anesthetics and pain medications used during delivery, (b) antibiotics and anti-bacterials used to cure infections, (c) drugs abused by pregnant women and (d) nutrients required for proper fetal growth. Placental transfer and exchange mechanisms are complicated processes, and in vitro models reflect only partially the equilibria that exist among mother, placenta and fetus. The perfused cotyledon model is elegant and simple but gives only restricted information. Isolated placental tissues give useful information about the pharmacological effects of drugs. Metabolic studies using human placental models provide information on the metabolism of a drug during placental transfer and accumulation of the drug or its metabolite in the placenta or fetal circulation. Several studies on the transplacental passage of drugs exist but many questions regarding the transfer of drugs between the maternal and fetal circulations and clearance of drugs from fetal circulation have yet to be answered. This article reviews in vitro and in vivo methods for evaluation of transplacental transport of drugs and their current effectiveness to obtain clinically useful data. PMID- 10837745 TI - Nutrient transport across the placenta. AB - The placenta forms a selective barrier that functions to transport nutrients that are of critical use to the fetus. Nutrient transport across the placenta is regulated by many different active transporters found on the surface of both maternal and fetal facing membranes of the placenta. The presence of these transporters in the placenta has been implicated in the facilitation of nutrient diffusion and proper fetal growth. In this review, recent developments concerning nutrient transporters that regulate glucose, amino acid, fatty acid, and nucleoside transplacental movement are discussed. PMID- 10837746 TI - Peptide transport and metabolism across the placenta. AB - Advances made in the field of DNA and recombinant technology have led to the emergence of peptides and proteins as an important class of therapeutic compounds. While a significant amount of information exists regarding the transport and metabolism of peptides across different barriers (e.g. gastro intestinal, nasal, and blood-brain barrier), limited attention has been paid towards their transport and metabolism across the placental barrier. The mechanism of placental transport of peptides is of importance in assessing the exposure of these drugs to the fetus, particularly when the drugs potentially may have adverse effects on the developing fetus.The absence of a well accepted, simple and convenient animal model may be a reason for the limited information available on the placental transport and metabolism of peptides. Although several in vivo models have been utilized to study the transport and metabolism of drugs across the placenta, species differences in the placental physiology and anatomy of the animal models with regard to the human placenta have prevented their widespread use. The in vitro human placental cell culture models are morphologically similar to the trophoblasts and often express the enzymes and carrier systems found in the human placenta. They can provide an easy and rapid method to determine the mechanisms of transport and metabolism of drugs across the placental barrier. These in vitro models have been utilized in the determination of transport mechanisms of drugs of abuse across the placenta.This article overviews the available literature on the placental transport of peptides and describes the application of an in vitro cell culture model (BeWo) to determine the mechanisms of transport of opioid peptides and their analogues across the placenta. PMID- 10837747 TI - Transplacental drug delivery: gene and virus delivery to the trophoblast. AB - The development of successful strategies for delivering genes to the placenta may provide new opportunities for modifying trophoblast function in order to learn more about trophoblast physiology and to offer novel therapeutic options for complications of pregnancy that result from placental dysfunction. Replication deficient recombinant viral vectors are useful vehicles for introducing genes into cells in vitro and in vivo. Recombinant adenovirus and herpes simplex virus vectors are unable to efficiently infect and transduce terminally differentiated trophoblastic cells. However, recombinant adeno-associated virus vectors transduce terminally differentiated trophoblastic cells, and a Sindbis virus construct efficiently transduces and destroys trophoblastic cancer cells. We describe the features that make particular viral vectors attractive for gene transfer to trophoblastic cells. PMID- 10837748 TI - The expression and regulation of drug metabolism in human placenta. AB - The human placenta oxidizes several xenobiotics, although the spectrum of substrates and metabolic activities when compared with the liver appears restricted. Maternal cigarette smoking or PCB exposure increase the expression of CYP1A1. This induced activity is able to catalyze the activation of benzo(a)pyrene into DNA-bound adducts, both in vitro and in vivo. Studies with RT PCR technique have demonstrated that first trimester placentae express at the mRNA level CYP1A1, 1A2, 2C, 2D6, 2E1, 2F1, 3A4, 3A5, 3A7 and 4B1 and at full term CYP1A1, 2E1, 2F1, 3A3/4, 3A5 and 3A7. However, more detailed studies on cDNA probes or with specific antibodies or 'diagnostic' substrates for other than CYP1A1, 2E1 and 3A gene products have yielded negative results. Studies on human placenta and a chorioncarcinoma cell line, JEG 3 cells, boulster the concept that placental CYP1A1 and 1B1 - although their expression is Ah receptor and ARNT mediated - is controlled by distinct mechanisms. Aromatase, CYP19, and cholesterol side-chain cleaving, CYP11B, genes, proteins and activities are catalytically active in human placentae throughout the pregnancy and those parameters do not seem to be affected by maternal cigarette smoking but rather maternal health status. However, the substrate binding pocket of aromatase accepts as its substrate several xenobiotics and is responsible for constitutive xenobiotic biotransformations.Functional placental glutathione S-transferase, N acetyl transferase and epoxide hydrolase are expressed via one gene each and their function reflects the placenta as an endocrine organ rather than a xenobiotic-metabolizing unit. However, markers for oxidative stress can be detected in decreased glutathione S-transferase activities.Because human placenta has quite well defined metabolic characteristics, and obtaining placental samples will not meet any drastic ethical difficulties, it could be used more intensively as a source of metabolizing enzymes in in vitro studies during the course of a drug development program. The human placenta, or its subcellular organelles, could serve as a real alternative model for an extrahepatic tissue in replacing recombinant expression systems especially if CYP11, 19, 1A1 or potentially 2E1 are target enzymes for potential metabolic interactions. PMID- 10837749 TI - Drugs of abuse and placental transport. AB - The placenta provides the only link between the mother and the developing fetus. The function of the placenta as a transport organ is obligatory for fetal development because this process, mediated by a variety of transport systems, is responsible for the delivery of nutrients from the mother to the fetus. Some of the transport systems in the placenta also play a role in the clearance of vasoactive compounds, thus maintaining optimal blood flow to this organ. There is strong supporting evidence to indicate that several of these placental transport systems are either direct or indirect targets for the abusable drugs cocaine, amphetamines, nicotine, and cannabinoids. These drugs of abuse compromise the placental transport function and consequently produce detrimental effects on the developing fetus. PMID- 10837750 TI - Preface. PMID- 10837751 TI - Long-acting control of ectoparasites: a review of collar technologies for companion animals. AB - Collar technologies for the prolonged topical control of ectoparasites on companion animals are reviewed, beginning with the first vinyl collars to the most recent developments. Matrix collars, such as the vinyl collar, have been the most commonly used technology, in which the active agent is blended directly into the polymer itself. Reservoir technologies, in which an active agent is dispersed or dissolved in a vehicle which is, in turn, held in a solid or mesh-like casing, are also discussed. Mechanical systems are presented including ultrasonic devices which emit sound waves designed to disrupt the flea life cycle, pumps which topically deliver small quantities of a dissolved active agent, and chambered devices designed to physically entrap the flea inside the collar. The future directions for topical parasite control are considered in closing, reviewing in particular the advent of more potent compounds. PMID- 10837752 TI - Hormone containing growth promoting implants in farmed livestock. AB - Growth promoting implants have been used in the production of cattle and sheep for over 40 years. Implants improve growth rate (+10 to 30%), feed efficiency (+5 to 15%) and carcass leanness (+5 to 8%). The history of this technology is mainly one of optimizing dose and hormone combinations, although matrices to optimize delivery rates of hormones from implants has received some attention. Estrogens are the first requirement for the growth response and in combination with androgens, growth is further enhanced. Several implant matrices are used, affecting pay-out rate and delivery time. The delivery time of most compressed implants is approximately 120 days and reimplantation after 60-120 days gives an additional response. Blood concentrations of implant hormones are increased and there appears to be a threshold blood level below which a growth response is not observed. Several proposed mechanisms are reviewed. The somatotropic axis appears most plausible for estrogens. Androgens may occupy muscle corticosteroid receptors. Regulated and proper use of implants assures their safety. PMID- 10837753 TI - Pulsatile release from subcutaneous implants. AB - Pulsatile delivery of antigens and hormones from subcutaneous implants could have uses in the animal production and veterinary medicine. Development of single-shot vaccines which release both initial and booster antigen from a single administration and hormonal preparations that release in a similar manner to the natural secretion patterns are two areas with potential. Formulation approaches employed to produce subcutaneous implants with pulsatile release profiles are reviewed. PMID- 10837754 TI - Somatotropin delivery to farmed animals. AB - Bovine somatotropin (bST) is marketed worldwide for increased milk production in cows while porcine somatotropin (pST) is approved in one country for increasing growth in swine. Somatotropin physicochemical properties, animal production method limitations and the need for cost effectiveness each contribute to the type of formulation developed. Various somatotropin physicochemical properties made formulation design difficult: heat and enzyme lability, tendency to aggregate, pH dependent solubility and stability, complicated degradation pathways and rapid in vivo clearance. The main problem of improving chemical and physical stability has been partially solved using certain excipients and vehicles. Formulations design to prolong somatotropin release include implants (matrix, osmotic), oleaginous suspensions and microparticles. This article presents the current status of somatotropin delivery in farmed animals, reviews the challenges encountered with formulation development, summarizes formulation approaches and discusses future somatotropin uses. PMID- 10837755 TI - Vaccine delivery to animals. AB - For many years vaccination of animals has been practiced to prevent infectious diseases using inactivated organisms or modified live organisms. The live vaccines were effective but lacked safety. The vaccines made with inactivated organisms required an adjuvant to induce an immune response that was not as effective as either the clinical disease or live vaccines. An 'ideal' vaccine would induce effective immunity specific for the type of infection, have long duration, require minimal or no boosters, have impeccable safety, would not induce adverse reactions, and be easy to administer. The desire to meet these criteria, and especially safety, has resulted in the development of vaccines that do not depend on the use of the viable disease agent. The emphasis on subunit or inactivated vaccines that meet the desired criteria of a perfect vaccine has resulted in a critical need for better adjuvants and delivery systems. This has resulted in a technological innovation revolution with development of a wide array of different technologies to generate effective vaccines. This review will describe the historical relevance of adjuvants used for parenterally administered inactivated/subunit vaccines as well as describe some of the exciting technological advances including adjuvants (ISCOMS), delivery systems (recombinant vectors, microparticles), and novel approaches (transgenic plants, naked DNA) that are currently being, or will be used in the future, in the search for better, more effective vaccines that meet the current and future needs of veterinary medicine. PMID- 10837756 TI - Membrane destabilization for improved cystolic delivery. PMID- 10837757 TI - Membrane fusion. AB - Although catalyzed by different proteins, the energy barriers for lipid bilayer fusion in exocytosis, viral fusion, and trafficking seem to be the same as those for the fusion of protein-free phospholipid membranes. To minimize this energy, fusion will proceed through a minimal number of lipid molecules, probably localized in bent non-bilayer intermediates. Experiments on phospholipid bilayer membrane fusion show the pathway of contact, hemifusion, flickering fusion pore formation, and fusion pore enlargement caused by swelling of the vesicle. Lipid curvature determines the barriers to hemifusion and fusion pore formation, while swelling-induced membrane tension drives fusion pore enlargement. Experiments on viral protein-induced cell-cell fusion and exocytosis show the same pathway with the same fundamental effects of lipid curvature and membrane tension. Thus while proteins control these reactions, lipid energetics determine the basic reaction scheme for membrane fusion. PMID- 10837758 TI - Lipid vesicles and membrane fusion. AB - Membrane fusion is essential for cell survival and has attracted a great deal of both theoretical and experimental interest. Fluorescence (de)quenching measurements were designed to distinguish between bilayermerging and vesicle mixing. Theoretical studies and various microscopic and diffraction methods have elucidated the mechanism of membrane fusion. These have revealed that membrane proximity and high defect density in the adjacent bilayers are the only prerequisites for fusion. Intermediates, such as stalk or inverse micellar structures can, but need not, be involved in vesicle fusion. Nonlamellar phase creation is accompanied by massive membrane fusion although it is not a requirement for bilayer merging. Propensity for membrane fusion is increased by increasing the local membrane disorder as well by performing manipulations that bring bilayers closer together. Membrane rigidification and enlarged bilayer separation opposes this trend. Membrane fusion is promoted by defects created in the bilayer due to the vicinity of lipid phase transition, lateral phase separation or domain generation, high local membrane curvature, osmotic or electric stress in or on the membrane; the addition of amphiphats or macromolecules which insert themselves into the membrane, freezing or other mechanical membrane perturbation have similar effects. Lowering the water activity by the addition of water soluble polymers or by partial system dehydration invokes membrane aggregation and hence facilitates fusion; as does the membrane charge neutralization after proton or other ion binding to the lipids and intermembrane scaffolding by proteins or other macromolecules. The alignment of defect rich domains and polypeptides or protein binding is pluripotent: not only does it increase the number of proximal defects in the bilayers, it triggers the vesicle aggregation and is fusogenic. Exceptions are the bound molecules that create steric or electrical barriers between the membranes which prevent fusion. Membrane fusion can be non-leaky but it is very common to lose material from the vesicle interior during the later stages of membrane unification, that is, after a few hundred microseconds following the induction of fusion. PMID- 10837759 TI - Role of the N-terminal peptides of viral envelope proteins in membrane fusion. AB - Membrane fusion is an important biological process that is observed in a wide variety of intra and intercellular events. In this review, work done in the last few years on the molecular mechanism of viral membrane fusion is highlighted, focusing in particular on the role of the fusion peptide and the modification of the lipid bilayer structure. While the Influenza hemagglutinin is currently the best understand fusion protein, there is still much to be learned about the key events in enveloped virus fusion reactions. This review compares our current understanding of the membrane fusion activity of Influenza and retrovirus viruses. We shall be concerned especially with the studies that lead to interpretations at the molecular level, so we shall concentrate on model membrane systems where the molecular components of the membrane and the environment are strictly controlled. PMID- 10837760 TI - To fuse or not to fuse: the effects of electrostatic interactions, hydrophobic forces, and structural amphiphilicity on protein-mediated membrane destabilization. AB - The development of lipid-based delivery vehicles for therapeutic molecules has become a topic of intense research. Recently, much of this effort has been directed towards mimicking the characteristics of viruses that give them an advantage for the delivery of genetic medicines. One of the most desirable properties of viral-based vectors is the ability to promote the destabilization of the host cell membrane to allow the entry of the genetic medicine into the target cell. This has been found to be largely controlled by the coat proteins on the surface of enveloped viruses. Although the exact mechanism by which proteins involved in the fusion process are able to promote the destabilization of membranes has yet to be elucidated, much understanding based upon information gained from a wide variety of studies is advancing the state of knowledge in this area. Parameters such as hydrophobic and electrostatic interactions as well as structural amphiphilicity, control to a large extent, the nature of the interaction of proteins with membranes. Thus, membrane fusion is mediated primarily by these forces acting in concert with one another. Ultimately, the knowledge gained from these studies will help to develop the ideal delivery system for the next generation of therapeutics. PMID- 10837761 TI - Application of membrane-active peptides for nonviral gene delivery. AB - A variety of membrane-modifying agents including pH-specific fusogenic or lytic peptides, bacterial proteins, lipids, glycerol, or inactivated virus particles have been evaluated for the enhancement of DNA-polycation complex-based gene transfer. The enhancement depends on the characteristics of both the cationic carrier for DNA and the membrane-modifying agent. Peptides derived from viral sequences such as the N-terminus of influenza virus haemagglutinin HA-2, the N terminus of rhinovirus HRV2 VP-1 protein, and other synthetic or natural sequences such as the amphipathic peptides GALA, KALA, EGLA, JTS1, or gramicidin S have been tested. Ligand-polylysine-mediated gene transfer can be improved up to more than 1000-fold by membrane-active compounds. Other polycations like dendrimers or polyethylenimines as well as several cationic lipids provide a high transfection efficiency per se. Systems based on these polymers or lipids are only slightly enhanced by endosomolytic peptides or adenoviruses. Electroneutral cationic lipid-DNA complexes however can be strongly improved by the addition of membrane-active peptides. PMID- 10837762 TI - Biological barriers to cellular delivery of lipid-based DNA carriers. AB - Although lipid-based DNA delivery systems are being assessed in gene therapy clinical trials, many investigators in this field are concerned about the inefficiency of lipid-based gene transfer technology, a criticism directed at all formulations used to enhance transfer of plasmid expression vectors. It is important to recognize that many approaches have been taken to improve transfection efficiency, however because of the complex nature of the formulation technology being developed, it has been extremely difficult to define specific carrier attributes that enhance transfection. We believe that these optimization processes are flawed for two reasons. First, a very defined change in formulation components affects the physical and chemical characteristics of the carrier in many ways. As a consequence, it has not been possible to define structure/activity relationships. Second, the primary endpoint used to assess plasmid delivery has been transgene expression, an activity that is under the control of cellular processes that have nothing to do with delivery. Gene expression following administration of a plasmid expression vector involves a number of critical steps: (i) DNA protection, (ii) binding to a specific cell population, (iii) DNA transfer across the cell membrane, (iv) release of DNA into the cytoplasm, (v) transport through the cell and across the nuclear membrane as well as (vi) transcription and translation of the gene. The objective of this review is to describe lipid-based DNA carrier systems and the attributes believed to be important in regulating the transfection activity of these formulations. Although membrane destabilization activity of the lipid-based carriers plays an important role, we suggest here that a critical element required for efficient transfection is dissociation of lipids bound to the plasmid expression vector following internalization. PMID- 10837763 TI - Cytosolic drug delivery using pH- and light-sensitive liposomes. AB - A growing body of literature describes the development and applications of novel targeting and/or contents release triggering schemes to improve the therapeutic index of drugs encapsulated within liposomes. This review focuses on literature appearing between January 1995-December 1997 that report 1) antibody and receptor mediated targeting approaches for improving drug localization and 2) acid, enzymatic, thermal or photochemical triggering processes that destabilize membranes and improve drug bioavailability via cytoplasmic delivery of liposomal contents. PMID- 10837764 TI - Preface. PMID- 10837765 TI - Investigation of distribution, transport and uptake of anti-HIV drugs to the central nervous system. AB - The distribution of currently available anti-HIV drugs into the CNS is reviewed with a focus on transport mechanisms. Among these drugs, nucleoside analogs are most well studied for their CNS distribution. The average reported values of the CSF/plasma steady-state concentration or corresponding AUC ratios are 0.23 (AZT), 0.06 (ddI), 0.04 (ddC), 0.49 (d4T), and 0.08 (3TC). Active efflux transport out of the CNS appears to be a predominant mechanism limiting nucleoside access to the CNS, although poor penetration may contribute to some extent for some polar nucleosides. The nature of the efflux pump for these drugs is speculated to be MRP-like transporter(s) in blood-brain and blood-CSF barriers. For non-nucleoside and protease inhibitors, much research remains to be done on the extent, time course, and mechanisms of their CNS distribution. The CNS penetration of some protease inhibitors is restricted by P-glycoprotein. A better understanding of transport mechanisms of anti-HIV drugs in the CNS is essential to develop approaches to enhance CNS delivery of available drugs and to identify new drugs less subject to active efflux transporter(s) in the CNS. PMID- 10837766 TI - Role of pharmacokinetics in the discovery and development of indinavir. AB - The discovery of indinavir is a successful example in which pharmacokinetic and metabolic information were incorporated into drug design. The use of animal and in vitro human metabolic data in predicting the oral bioavailability and hepatic clearance in humans was critical in selecting indinavir as a drug candidate for development. In its development stage, pharmacokinetics continued to play an important role in identifying the key properties of indinavir in vivo, which allowed the characterization and prediction of the time course of drug action under physiological and pathological conditions. This review describes the role of pharmacokinetics and drug metabolism in the discovery and development of indinavir. PMID- 10837767 TI - Mechanisms of transport of nucleosides and nucleoside analogues in choroid plexus. AB - Nucleoside and nucleobase analogues are being used to treat a number of viral infections of the central nervous system (CNS) including herpes-simplex encephalitis, cytomegalovirus retinitis, and AIDS-related dementia complex. Delivery of nucleoside analogues to the CNS is considered a key challenge in the treatment of these diseases. In this review, we focus on the mechanisms of transport of nucleosides and nucleoside analogues in the choroid plexus. First, the structure and function of the choroid plexus are reviewed. Then, we focus on the mechanisms of nucleoside transport in mammalian cells. Specific emphasis is placed on the molecular and functional characteristics of various nucleoside transporters. A discussion is then devoted to the mechanisms of nucleoside transport in choroid plexus. Current knowledge of nucleoside transport systems in choroid plexus in several animal species is summarized followed by a short section on the mechanisms of transport of nucleobases in the choroid plexus. Finally, a brief section on future directions is included. PMID- 10837768 TI - Concepts for the design of anti-HIV nucleoside prodrugs for treating cephalic HIV infection. AB - The life cycle of HIV involves nine sequential stages. Of these, the reverse transcription (RT) process is a prime target for drug therapy, using both nucleoside and non-nucleoside inhibitors of RT. There are currently five marketed 2',3'-dideoxynucleoside RT inhibitors, but there is need for drugs with improved therapeutic efficacy, decreased development of resistance and broader spectrum to treat resistant strains.One approach to improve RT inhibitors is through chemical derivatization using metabolically-cleavable linkages that permit timely regeneration of the active nucleoside inside the body at the site of infection (prodrug formation). Four classes of prodrugs are now reviewed: 2',3' dideoxynucleoside masked phosphates, 5'-O-carboxylic acid esters of 2',3' dideoxynucleosides, 2',3'-dideoxycytidine N(4)-[(dialkylamino)methylene] prodrugs and 5-halo-6-alkoxy(azido or hydroxy)-5,6-dihydro 2',3'-dideoxynucleosides. Mutually-masking dual action (MMDA) prodrugs that release a nucleoside RT inhibitor and an abnormal N-myristoyl transferase substrate are presented as a special class of anti-HIV prodrugs that have the potential to interact with the life cycle of the virus at two distinct stages. PMID- 10837769 TI - Transport, metabolism and elimination mechanisms of anti-HIV agents. AB - Currently available anti-HIV drugs can be classified into three categories: nucleoside analogue reverse transcriptase inhibitors, non-nucleoside reverse transcriptase inhibitors, and protease inhibitors. Knowledge of these anti-HIV drugs in various physiological or pharmacokinetic compartments is essential for design and development of drug delivery systems for the treatment of HIV infection. The input and output of anti-HIV drugs in the biological systems are described by their transport and metabolism/elimination in this review. Transport mechanisms of anti-HIV agents across various biological barriers, i.e., gastrointestinal wall, skin, mucosa, blood cerebrospinal barrier, blood-brain barrier, placenta, and cellular membranes, are discussed. Their fates during and after systemic absorption and their metabolism-related drug interactions are reviewed. Many anti-HIV drugs presently marketed in the US bear some significant drawbacks such as relatively short half-life, low bioavailability, poor penetration into the central nervous system, and undesirable side effects. Efforts have been made to design drug delivery systems for the anti-HIV agents to: (1) reduce the dosing frequency; (2) increase the bioavailability and decrease the degradation/metabolism in the gastrointestinal tract; (3) improve the CNS penetration and inhibit the CNS efflux; and (4) deliver them to target cells selectively with minimal side effects. We hope to stimulate further interests in the area of controlled delivery of anti-HIV agents by providing current status of transport and metabolism/elimination of these agents. PMID- 10837770 TI - P-glycoprotein, secretory transport, and other barriers to the oral delivery of anti-HIV drugs. AB - Orally administered anti-HIV drugs must be adequately and consistently absorbed for therapy to be successful. This review discusses the barriers to achieving oral bioavailability for the currently available anti-HIV drugs. Most reverse transcriptase inhibitors have good oral bioavailabilities. Didanosine bioavailability could be reduced by acid instability, first-pass hepatic metabolism, and possibly poor intestinal permeation. Bioavailability of zidovudine is also reduced by first-pass metabolism. The non-nucleoside reverse transcriptase inhibitors have oral bioavailabilities most probably limited by poor aqueous solubility. For each of the currently marketed HIV protease inhibitors, solubility, intestinal permeability, and first-pass metabolism could contribute to reducing oral bioavailability. The intestinal permeabilities of these agents is influenced by secretory transport. In vitro, secretory transport, which appears to be P-glycoprotein-mediated, is much greater than permeation in the absorptive direction for indinavir, nelfinavir, ritonavir, and saquinavir. The mechanisms of secretory intestinal transport are reviewed, and the factors that may influence the impact of secretory transport in vivo are considered. PMID- 10837771 TI - Development and optimization of anti-HIV nucleoside analogs and prodrugs: A review of their cellular pharmacology, structure-activity relationships and pharmacokinetics. AB - Significant improvements in antiviral therapy have been realized over the past 10 years. Numerous nucleoside analogs, as well as prodrugs of active compounds, have been synthesized and tested for anti-HIV activity. In addition to the five nucleoside analogs currently used clinically for the treatment of HIV infection, a broad spectrum of anti-HIV nucleoside analogs (including 2',3' dideoxynucleoside analogs, oxathiolanyl 2',3'-dideoxynucleoside analogs, dioxolanyl 2',3'-dideoxynucleoside analogs, carbocyclic 2',3'-dideoxynucleoside analogs and acyclic nucleoside analogs) and their prodrugs (including ester prodrugs, phospholipid prodrugs, dihydropyridine prodrugs, pronucleotides and dinucleotide analogs), targeted at HIV reverse transcriptase, are reviewed with focus on structure-activity relationships, cellular pharmacology and pharmacokinetics. Several of these anti-viral agents show promise in the treatment of AIDS. PMID- 10837772 TI - Studies of macromolecular prodrugs of zidovudine. AB - The current problems in controlling severe viral infections such as AIDS as well as the lack of effective and safe therapeutic measures for such diseases have caused interest in systems such as macromolecular prodrugs potentially able to solve heavier drawbacks of conventional antiviral therapy. This review focuses on various approaches proposed in the literature in this field. Neoglycoproteins and synthetic protein-like structure polymers have been mainly proposed. In the first group, the possibility of incorporating into the polymeric structures a determined amount of sugar molecules make them interesting candidates for targeting of infected blood cells. The conjugate of zidovudine (AZT) and an anti transferrin receptor antibody OX-26 has been proposed for brain targeting. The conjugate of AZT with alpha,beta-poly(N-hydroxyethyl)-DL-aspartamide (PHEA) showed good release properties in a prolonged time. PMID- 10837773 TI - Improved uptake and retention of lipophilic prodrug to improve treatment of HIV. AB - Dideoxynucleosides currently in use for anti-HIV therapy have been found to be inefficient in passing through the blood-brain barrier to enter and maintain therapeutic drug levels in brain, a very significant reservoir of HIV. The low bioavailability of these drugs combined with the bone marrow toxicity of AZT (3' azido, 3'-deoxythymidine, Zidovudine), resulting in anemia and leukopenia, pancreatitis with ddI (2',3'-dideoxyinosine, Didanosine) and painful peripheral neuropathy in case of ddC (2',3-dideoxycytosine, Zalcitabine) are the limiting factors in their use. In addition, the emergence of strains of HIV resistant to AZT, the most commonly used drug, further restricts its use. Thus the control of AIDS and its complications, needs special therapeutic approaches to combat the disease. In order to overcome these limitations, AZT and ddI have been synthesized as ester-linked ceramide- and phosphatidylcholine-linked prodrugs possessing therapeutic attributes lacking in the parent compounds. There is greater uptake and longer retention of these prodrugs in NIH/3T3 cells in vitro. Pretreatment with our prodrugs blocked infection of these cells by Moloney murine leukemia virus (M-MuLV) for an extended period, which the parent drugs failed to do. When human CD4+ HeLa cells were continuously exposed to the AZT prodrug, subsequent infection of these cells by HIV was blocked. Similar results were obtained with NIH/3T3 cells exposed to M-MuLV. AE(6)C, a prodrug of AZT linked to ceramide via a cleavable ester bond and a six carbon linker, was less toxic to both mouse and human bone marrow progenitor cells than free AZT. Most significantly, the prodrugs concentration was greater and the retention longer, in well known sanctuaries for HIV, such as the brain, testes and thymus. PMID- 10837774 TI - Involvement of multiple transporters in the oral absorption of nucleoside analogues. AB - Many nucleoside analogues such as azt, ddI, ddC, d4T, 3TC, acv and vacv are currently being used in the treatment of patients infected with HIV, suffering from AIDS, or AIDS-related opportunistic infections. The transport of nucleoside analogues across the gastrointestinal tract is mediated by a number of transporters that fall into three broad categories, i.e., Na(+)-dependent concentrative transporters, Na(+)-independent equilibrative transporters and H(+)/peptide transporters. The first two transporter classes contain a large number of subtypes that are based on the substrate specificity. Recent studies have shown that most of the anti-HIV nucleoside analogues are transported by one or more of the nucleoside transporters. Furthermore, certain analogues, such as acv, appear to be absorbed by non-carrier-mediated diffusion, whereas vacv is apparently transported by non-nucleoside transporters (e.g., the oligopeptide transporter, PepT1 and possibly others). Thus, it is desirable to understand the precise nature of the absorption mechanism of these drugs to improve bioavailability and reduce the variability that is commonly observed in vivo in human patients. A complete understanding of the complex interactions of nucleoside analogues with the various transporters will help in designing better delivery systems and strategies to improve efficacy. In the current report, the mechanisms of nucleoside and nucleoside-analogue transport are reviewed. Also, methods of exploiting prodrugs to improve the bioavailability characteristics of drugs are highlighted. PMID- 10837775 TI - Oral absorption of the HIV protease inhibitors: a current update. AB - Although the human immunodeficiency virus (HIV) protease inhibitors are highly effective, they are characterized by low and/or variable bioavailability with limited penetration into the central nervous system (CNS). Their clinical use is limited by patient compliance and by drug-drug interactions. The effect of drug solubility on their oral absorption has been investigated but further evaluation of this relationship is required. First pass metabolism appears to be significant for the HIV protease inhibitors and they are extensively metabolized by cytochrome P450 (CYP) 3A4. Recent studies suggest that these drugs are substrates for the P-glycoprotein efflux pump, which can limit their intestinal absorption and their transport across the blood-brain barrier. Drugs inducing or inhibiting CYP3A4 and/or P-glycoprotein may influence the bioavailability of the HIV protease inhibitors. The low bioavailability, variable absorption and drug-drug interactions of the HIV protease inhibitors may be related to the variability of cytochrome P450 and P-glycoprotein expression and to possible CYP3A4/P glycoprotein interactions. To improve oral HIV protease inhibitor therapy, it is essential to mechanistically characterize the cell specific, tissue specific and regional intestinal dependencies of drug transport, secretory transport, metabolism and P-glycoprotein/CPY3A4 interactions. This report reviews the physicochemical characteristics and pharmacokinetics of the HIV protease inhibitors while considering the relationships between their hepatic and intestinal metabolism, low bioavailability, variable absorption and drug-drug interactions. PMID- 10837776 TI - The Pharmacokinetics of Liposomes in Tumor Targeting. PMID- 10837777 TI - Mechanisms and kinetics of liposome-cell interactions. AB - Although the possibility of targeting drugs to specific tissues and cells, as well as facilitating their uptake and cytoplasmic delivery has rendered liposomes a versatile drug carrier system with numerous potential applications in medicine, the molecular mechanisms of liposome-cell interactions are not understood well. Here we have reviewed the early and current concepts of liposome-cell interactions, including possible liposome receptors. Uptake of liposomes by cells can be modified by the lipid composition, particularly by the inclusion of steric stabilizers such as PEG-conjugated lipids. Such modifications also alter the circulation time and biodistribution of liposomes, which can thus be tailored for particular applications. The intracellular fate of encapsulated molecules can be modified by the use of pH-sensitive liposomes which can also be sterically stabilized. Cationic liposomes that can undergo lipid mixing with cellular membranes can deliver complexed DNA to cells, but most likely via an endocytotic process. Kinetic analysis of liposome-cell interactions can elucidate the numbers of liposome receptors of several types and the corresponding binding constants. It is likely that liposomes bind to different cell surface receptors on different cells, and that they utilize more than one type of receptor on a particular cell. The kinetic analysis also provides the rate constants of endocytosis and the percentages of liposomes that are bound or endocytosed. PMID- 10837778 TI - Pharmacokinetics of anticancer drugs, plasmid DNA, and their delivery systems in tissue-isolated perfused tumors. AB - To achieve an optimal chemotherapy or gene therapy against tumors or to realize rational design of delivery systems for cancer therapy, pharmacokinetic information in tumor should be obtained. A tissue-isolated tumor preparation is a useful experimental system to investigate the intratumoral disposition of drugs, carriers, and their complexes. The disposition of drugs in the solid tumor was analyzed in this system after intraarterial infusion (systemic route) or by intratumoral injection (topical route). Here the results of low-molecular weight drugs, their macromolecular prodrugs, lipid carriers like fat emulsions and liposomes, and plasmid DNA and its complexes, are addressed. Pharmacokinetic analyses in the tumor clearly indicate that the intratumoral fate of drugs and delivery systems are determined by (i) the anatomical and physiological properties of the tissue and (ii) the physicochemical characteristics of drugs and delivery systems such as molecular weight, size, lipophilicity, and electrical charge. These approaches are useful for designing and developing optimized drug delivery systems. PMID- 10837779 TI - Pharmacokinetic/pharmacodynamic modeling of antitumor agents encapsulated into liposomes. AB - Pharmacokinetic/pharmacodynamic (PK/PD) modeling of antitumor agents has been developed for doxorubicin (DOX) in order to predict the optimum conditions for a drug carrier to maximize the antitumor effect. A PK model was constructed for free and liposomal doxorubicin using a hybrid model wherein the disposition in the whole body is described by compartment models, which were linked to the tumor compartment via the blood flow rate. The PD model for doxorubicin was described by a cell-kill kinetic model, which represents the number of tumor cells quantitatively, as a function of the free concentration of doxorubicin in the tumor compartment. The influence of each parameter on the antitumor effects was examined by sensitivity analysis based on the PK/PD model, which clearly showed the importance of optimizing the release rate of DOX from liposomes. The validity of the model has been tested using animal experiments. Preliminary simulations were also performed for humans after scaling up the PK/PD model from rodents to humans. The optimum conditions in the rate of drug release from liposomes were different for rodents vis-a-vis humans, which indicates the limitations involved in extrapolating optimum conditions for experimental animals to those for humans. PMID- 10837780 TI - Anticancer therapy using glucuronate modified long-circulating liposomes. AB - Since conventional liposomes tend to be trapped by the reticuroendothelial systems (RES), their use as drug carriers is limited when the targets are not RES cells. Therefore, many attempts have been made to avoid the RES-trapping of liposomes. Favorable results were obtained by a modification of liposomes with a glucuronic acid derivative, PGlcUA, and polyethyleneglycol. These liposomes have a long-circulating character, and showed the further advantage for passive targeting to tumor tissues, since the vasculature in tumor tissues is leaky enough for small-sized liposomes to extravasate. Thus long-circulating liposomes are useful for tumor imaging and treatment. PGlcUA-modified liposomes were actually found to accumulate effectively in tumor tissue, and showed enhanced efficacy of antitumor agents, such as adriamycin and vincristine when they were encapsulated into the liposomes. Usefulness of PGlcUA liposomes as drug carriers was also observed in photodynamic therapy and in treatment of cancer by amphiphilic novel antitumor agents. PMID- 10837781 TI - The size of liposomes: a factor which affects their targeting efficiency to tumors and therapeutic activity of liposomal antitumor drugs. AB - The size of liposomes has been shown to be an important factor in the efficient delivery of an antitumor agent to a tumor. In this paper, the effects of the size of liposomes on the pharmacokinetics of liposomes and liposome-encapsulated drugs are discussed with reference to: (1) the circulation amount and residence time of liposomes in the blood, (2) the accumulation of liposomes in the tumor, and (3) in vivo drug release from liposomes. In addition, the effect of size on therapeutic activity (antitumor efficacy and toxicity) of a liposomal anticancer preparation is discussed. Finally we discuss the importance of liposome size in the design of a more effective liposomal antitumor preparation. PMID- 10837782 TI - Possibility of active targeting to tumor tissues with liposomes. AB - In terms of active targeting by immunoliposomes, two anatomical compartments are considerable for targeting sites. One is located a readily accessible site in intravascular, and another is a much less accessible target site located in the extravascular. However, it was made clear that the active targeting with immunoliposomes is determined by two kinetically competing processes, such as binding to the target site and uptake by the RES. To overcome these contradictions, we have designed a new type of long-circulating immunoliposome, which was PEG-immunoliposome attached antibodies at the distal end of PEG chain, so called the pendant type immunoliposome. The pendant type immunoliposome showed much higher targetability than the ordinary immunoliposomes to both targeting sites of lung endothelial cells and solid tumor tissue. This is due to the free PEG chains (not linked to the antibody) effectively avoiding the RES uptake of liposomes, resulting in elevated the blood concentration and enhanced the target binding of immunoliposomes. The presence of free PEG does not interfere with the binding of the terminally linked antibody to the antigen. For targeting to the vascular endothelial surface in the lung, 34A antibody, which is highly specific to mouse pulmonary endothelial cells, was conjugated to make the pendant type immunoliposomes (34A-PEG-ILP). 34A-PFG-ILP showed significantly higher targeting degree than the ordinary type of immunoliposomes. For targeting to the solid tumor tissue, Fab' fragment of 21B2 antibody which is anti-human CFA and transferrin (TF) were used. Both pendant type immunoliposomes (Fab'-PFG-ILP and TF-PEG-ILP) showed the low RES uptake and the long circulation time, and resulted in enhanced accumulation of the liposomes in the solid tumor. TF-PEG-ILP was internalized into tumor cells with receptor mediated endocytosis, after extravasation into tumor tissue. The pendant type immunoliposome can escape from the gaps between adjacent endothelial cells and openings at the vessel termini during tumor angiogenesis by passive convective transport much rather than ligand directed targeting. Active targeting to tumor tissue with the pendant type immunoliposome is particularly important for many highly toxic anticancer drugs for cancer chemotherapy. An ultimate goal of pendant type immunoliposome is the incorporation of a fusogenic molecule that would induce fusion of liposome following their binding to the target cells or their internalization by endocytosis. Such liposomal formulations should be useful for endocytotic internalization of plasmid DNA and other bioactive materials. PMID- 10837783 TI - Immunoliposomes for the targeted delivery of antitumor drugs. AB - This review presents an overview of the field of immunoliposome-mediated targeting of anticancer agents. First, problems that are encountered when immunoliposomes are used for systemic anticancer drug delivery and potential solutions are discussed. Second, an update is given of the in vivo results obtained with immunoliposomes in tumor models. Finally, new developments on the utilization of immunoliposomes for the treatment of cancer are highlighted. PMID- 10837784 TI - Preface PMID- 10837785 TI - Blood substitutes. AB - Red cell substitutes are a group of oxygen carriers designed to temporarily replace transfused blood. Each product is unique in its limitations and advantages. Research and development has been slow because of the far-reaching consequences of replacing an oxygen carrier outside of the red cell. Nevertheless, a number of products are in advanced clinical trials and nearing the market. When they are available for use it is likely that development will accelerate and even better products will substantially alleviate the world-wide shortage of blood for transfusion and enable the delivery of medical care to underserved populations. An important consequence of the development of these products has been a better understanding of how oxygen is delivered to tissues. PMID- 10837786 TI - Current status of artificial oxygen carriers. AB - Artificial oxygen carriers may be grouped into modified hemoglobin solutions and fluorocarbon emulsions. In animal experiments, both have been shown to be efficacious in improving tissue oxygenation and as substitutes for blood transfusions. Advantages and disadvantages are being discussed in this article as well as the latest steps in the clinical development. PMID- 10837787 TI - Purification and chemical modifications of hemoglobin in developing hemoglobin based oxygen carriers. AB - The efficacy of blood substitutes, as a whole, has been readily demonstrated, in animals as well as clinical studies. It is well known that stroma free hemoglobin (SF-Hb) is very toxic, due to effects on renal and coagulation functions and vascular tone. Several modifications have been made to the hemoglobin tetramer in an attempt to eliminate its toxicity. Conjugation, cross-linking, polymerization, and recombinant technology have all been used to reduce toxicity, while aiming to optimize the therapeutic value of hemoglobin based blood substitutes. The remaining issue seems to be the hypertensive response seen in many hemoglobin solutions. The cause of the hypertensive response, and hence what chemical modifications are suitable to alleviate it are still under debate. PMID- 10837788 TI - Tissue perfusion and oxygenation with blood substitutes. AB - As an alternative to transfusion of red blood cells, intravenously (iv) administered artificial oxygen (O(2)) carriers are intended to increase the reduced O(2) carrying capacity of blood in the case of acute severe anemia, i.e. hemorrhagic shock or extreme normovolemic hemodilution (ANH). Actually, two groups of artificial O(2) carriers are investigated: ultrapurified, stroma-free hemoglobin solutions (SFH) of human or bovine origin and synthetically produced perfluorocarbons (PFC). SFH may be administered in large amounts and are suitable for 1:1 replacement of blood losses in case of hemorrhage as well as for isovolemic exchange of blood during ANH. In both situations SFH solutions effectively restore (hemorrhagic shock) and maintain (extreme ANH) tissue oxygenation despite extremely low hematocrit values. The vasopressor property of the isolated Hb molecule leads to a species-dependent (rodent>pig>human) increase in systemic and pulmonary vascular resistance, but leaves overall distribution of cardiac output uninfluenced. Due to the particulate nature of PFC emulsions, iv administration has to be restricted to small doses (3-4.5 ml/kg body weight for the actually investigated 60% w/v perflubron emulsion) in order to avoid overload of the reticuloendothelial system. Thus PFC emulsions are unsuitable for isovolemic blood replacement in hemorrhagic shock or ANH. Low-dose iv PFC administration in already hemodiluted subjects, however, creates an additional margin of safety to guarantee adequate tissue oxygenation which allows for further, extreme ANH, without risking tissue hypoxia. PMID- 10837789 TI - Current aspects in pharmacology of modified hemoglobins. AB - Blood substitutes are products that are designed to replace whole blood (or) red blood cells in the field of transfusion medicine. There are two major classes that belong to this new therapeutics: (i) modified hemoglobins and (ii) perfluorocarbons. Modified hemoglobins have made tremendous progress in the past decade and are being considered for a wide variety of conditions like trauma, elective surgery, oxygenation of tumors to make them more sensitive to radiation therapy, stroke etc. Although, these agents are primarily used for oxygen delivery, their pharmacological actions have been significantly important. Several mechanisms are being explored to explain these pharmacological effects. Modified hemoglobins suffer several drawbacks including hypertension, renal toxicity, and pulmonary hypertension that restrict their development. This review deals with the clinical status and pharmacological actions of modified hemoglobins presently in advanced stages of development and some of the newer generation hemoglobin based therapeutics are also discussed. PMID- 10837790 TI - Site-specific modifications and toxicity of blood substitutes. The case of diaspirin cross-linked hemoglobin. AB - Safe and effective hemoglobin-based blood substitutes may be advantageous over conventional therapies for certain clinical settings requiring short term blood replacement such as emergency resuscitation and hemodilution in surgery. Many advances have been made in developing these oxygen therapeutics, however safety concerns continue to slow their clinical progress. An important and often overlooked consideration in evaluating the safety of modified hemoglobins is the impact of chemical and/or genetic modifications on the redox chemistry of these proteins. Diaspirin cross-linked hemoglobin (DBBF-Hb) has been extensively evaluated in vitro and in animal models, and thus represents a useful model to explore possible correlations between structural-functional alterations and toxicity of hemoglobin-based blood substitutes. PMID- 10837791 TI - Two future generations of blood substitutes based on polyhemoglobin-SOD-catalase and nanoencapsulation. AB - This paper discusses our research on two new generation blood substitutes. One is based on the crosslinking of hemoglobin, superoxide dismutase(SOD) and catalase (CAT) to form polyhemoglobin-SOD-CAT. This is being investigated for use in conditions with potential problems related to ischemia-reperfusion injuries as in severe hemorrhagic shock, stroke and other conditions. The second one is based on biodegradable polymeric nanocapsules containing hemoglobin and enzymes. In this form, the hemoglobin and enzymes are separated from the external environment. Furthermore, modifications of the polymeric membrane can result in increase in circulation time. PMID- 10837792 TI - Nerve regeneration in Wld(s) mice is normalized by actinomycin D. AB - Injured nerves of Wld(s) mice neither degenerate nor regenerate for several weeks. We have conjectured that Wld(s) axons have the ability to regenerate but its expression is impaired by the Schwann cells of the undegenerated distal stump. To test this conjecture, transcription was locally arrested with actinomycin D (ActD), nerves were crushed, and regrowth was evaluated. In normal CD1 nerves injected with ActD 3 days before the crush, the rate of elongation was not affected but the delay of regrowth was shortened. In sharp contrast, ActD normalized the elongation of Wld(s) axons. When Wld(s) nerves were crushed past the treated segment, axons did not regenerate. After 7, but not 4, days of treatment, intact CD1 and Wld(s) axons presented a local sprouting response. We conclude that Wld(s) axons can regenerate in a normal way but do not do so because the undegenerated Schwann cells of the distal stump repress the regrowth program. We present a model axon that includes a destruction program and a post transcriptional trophic regulation of its phenotype. PMID- 10837793 TI - Phospholipid composition and levels are altered in Down syndrome brain. AB - Phospholipid composition (mol %) and levels (nmol/mg protein) were determined in postmortem frontal cortical and cerebellar gray matter from older Down Syndrome (DS) patients (age range 38-68 years) and from control subjects. Neither DS nor control tissue exhibited any age-dependent alteration in phospholipid composition or levels. Total phospholipid content was significantly reduced approximately 20% in DS frontal cortex and cerebellum relative to these regions in control tissue. Individual phospholipid levels were also reduced in DS frontal cortex and cerebellum, including a specific 37% decrease in phosphatidylinositol (PtdIns) and a nearly 35% decrease in ethanolamine plasmalogen. Because of the large decrease in phospholipid content in DS brain, the cholesterol/phospholipid ratio was calculated for each group. There was no significant difference in this ratio between groups, indicative of compensatory changes to keep the cholesterol/phospholipid ratio constant. Despite the large changes in DS brain phospholipid levels, significant changes in composition were limited to a 18% decrease in PtdIns mol % and a 22% increase in the mol % of sphingomyelin. These results suggest either a decrease in membrane phospholipids due to a loss of dendrites and dendritic spines, or a general defect in brain lipid metabolism in older DS subjects. The proportionally greater alterations in PtdIns and PlsEtn levels, indicate that the metabolism of these two phospholipids was affected to a greater extent than the other phospholipids. Further, because these changes are found in both the frontal cortical and cerebellar gray matter, they likely are related to the Down syndrome condition rather than to Alzheimer neuropathology. PMID- 10837794 TI - Long-term effects on feeding and body weight after stimulation of forebrain or hindbrain CRH receptors with urocortin. AB - Research on the contribution of CRH receptor stimulation to energy homeostasis has focused on forebrain substrates. In this study, we explored the effects of caudal brainstem administration of the CRH receptor agonist, urocortin, on food intake and body weight, and on plasma glucose and corticosterone (CORT) in non deprived rats. Urocortin (0, 0.3, 1, 3 microg) delivered, respectively, to the fourth and lateral ventricles yielded substantial suppression of food intake measured 2, 4 and 24 h later. A significant but more modest anorexia was observed between 24 and 48 h after injection. Intake responses did not differ between the injection sites, but body weight loss measured 24 h after lateral-i.c.v. injection was substantially greater than that after fourth-i.c.v. injection. Fourth-i.c.v. urocortin administration (3 microg) produced substantial elevations in plasma glucose and CORT that were not distinguishable in magnitude and duration from responses to lateral-i.c.v. delivery. Unilateral microinjection of urocortin into the dorsal vagal complex significantly reduced 24-h food intake at a dose (0.1 microg) that was subthreshold for the response to ventricular administration, suggesting that fourth-i.c.v. effects are mediated in part by stimulation of CRH receptors in this region of the caudal brainstem. The results indicate that similar effects can be obtained from stimulation of anatomically disparate populations of CRH receptors, and that interactions between forebrain and hindbrain structures should be considered in the evaluation of CRH contributions to food intake and body weight control. PMID- 10837795 TI - An animal model of adolescent nicotine exposure: effects on gene expression and macromolecular constituents in rat brain regions. AB - Nearly all smokers begin tobacco use in adolescence, and approximately 25% of US teenagers are daily smokers. Prenatal nicotine exposure is known to produce brain damage, to alter synaptic function and to cause behavioral anomalies, but little or no work has been done to determine if the adolescent brain is also vulnerable. We examined the effect of adolescent nicotine exposure on indices of cell damage in male and female rats with an infusion paradigm designed to match the plasma levels found in human smokers or in users of the transdermal nicotine patch. Measurements were made of DNA and protein as well as expression of mRNAs encoding genes involved in differentiation and apoptosis (p53, c-fos) in cerebral cortex, midbrain and hippocampus. Following nicotine treatment from postnatal days 30 47.5, changes in macromolecular constituents indicative of cell loss (reduced DNA) and altered cell size (protein/DNA ratio) were seen across all three brain regions. In addition, expression of p53 showed region- and gender-selective alterations consistent with cell damage; c-fos, which is constitutively overexpressed after gestational nicotine exposure, was unaffected with the adolescent treatment paradigm. Although these measures indicate that the fetal brain is more vulnerable to nicotine than is the adolescent brain, the critical period for nicotine-induced developmental neurotoxicity clearly extends into adolescence. Effects on gene expression and cell number, along with resultant or direct effects on synaptic function, may contribute to increased addictive properties and long-term behavioral deficits. PMID- 10837796 TI - Substantia nigra pars reticulata lesion induces preconvulsive behavior and changes in glutamate receptor gene expression in the rat brain. AB - The substantia nigra pars reticulata (SNpr) has been proposed to play an important role in the control of the propagation and/or the generation of epileptic seizures. Earlier studies have shown differential effects of the lesion of the SNpr on seizure genesis that demonstrated a regional difference in the anterior and posterior parts of the SNpr in preconvulsive behavior induced by unilateral reticulata injection of dopamine (DA). This study was aimed to investigate some of the underlying mechanisms of the preconvulsive behavior elicited by unilateral SNpr DA injection by the study of changes in the gene expression of glutamate receptor subunits (GluR1, GluR2 and NMDAR1) and of changes in animal behavior following coinfusion of DA and a DA D1 antagonist SCH 23390 into the SNpr. Unilateral injection of exogenous DA into the anterior region of the SNpr induced rapid and short lasting preconvulsive behavior up to wet dog shakes stage and a significant reduction of gene expression for GluR1, GluR2 and NMDAR1 subunits in rat hippocampal subfields including CA1 through CA4 and dentate gyrus (DG) at 1 day after nigral DA injection. The effect was long lasting and persisted for at least 3 weeks. Both preconvulsive behavior and downregulation of glutamate receptor subunit genes were completely blocked by simultaneous coinfusion of DA and SCH 23390. The results suggest, for the first time, that DA D1 receptor in the SNpr may mediate the nigral-involved seizure development. Glutamate desensitization, and/or selective early neuronal damage might be responsible for the downregulation of glutamate receptor subunits by transient preconvulsive activity. PMID- 10837797 TI - Noradrenergic lesions differentially alter the expression of two subtypes of low Km cAMP-sensitive phosphodiesterase type 4 (PDE4A and PDE4B) in rat brain. AB - This study examined the effects of selective, central noradrenergic dennervation with 6-hydroxydopamine (6-OHDA) on the expression of type 4 phosphodiesterases (PDE4). Twenty-one days following i.c.v. injection of 6-OHDA (200 microg) hypothalamus, neostriatum, and cerebellum were dissected. Infusion of 6-OHDA reduced norepinephrine (NE) content in all the brain areas examined (to 17%, 76% and 16% of sham-operated controls in hypothalamus, striatum, and cerebellum, respectively). 6-OHDA injections also reduced dopamine levels in hypothalamus (53%) and neostriatum (68%). Administration of desipramine (20 mg/kg, i.p.) 30 min prior to 6-OHDA injection protected neostriatal and cerebellar noradrenergic neurons NE levels (110-122% of the control levels). Desipramine partially attenuated the 6-OHDA-mediated decrease in NE content of hypothalamus, but had little or no effect on either striatal or hypothalamic dopamine (DA) levels. Western blot analysis using a PDE4A-selective antibody revealed three major bands (109 kDa PDE4A5, 102 kDa PDE4AX and 76 kDa PDE4A1) in hypothalamus and striatum. Infusion of 6-OHDA decreased the expression of PDE4A5 and PDE4AX but not of PDE4A1 in hypothalamus, as determined by quantitative Western blotting. Pretreatment of rats with desipramine attenuated the 6-OHDA-induced down regulation of PDE4A5 and PDE4AX bands in hypothalamus. The PDE4B selective antibody K118 labels 5 major bands in all the brain regions studied. One hundred kDa PDE4B3, 86 kDa PDE4B2 and a 78 kDa PDE4B band was identified using recombinant proteins. Treatment of rats with 6-OHDA resulted in a 52% decrease in the PDE4B3 and 58% decrease in 78 kDa PDE4B variant in hypothalamus; administration of desipramine attenuated the 6-OHDA-induced down-regulation of both PDE4B variants. Neither 6-OHDA nor desipramine altered striatal PDE4A or PDE4B isozymes. In contrast, cerebellar PDE4B3 variant is up-regulated by 6-OHDA treatment and were partially normalized to control values by desipramine pretreatment. These data demonstrate that PDE4 subtypes are differentially regulated by presynaptic noradrenergic activity and may play an important role in the maintaining homeostasis of noradrenergic signal transduction in rat brain. PMID- 10837798 TI - Caspase inhibitors are functionally neuroprotective against oxygen glucose deprivation induced CA1 death in rat organotypic hippocampal slices. AB - We have explored the neuroprotective efficacy of the cell penetrant caspase inhibitor, Ac-YVAD-cmk, in a hippocampal slice model of neuronal cell death induced by oxygen and glucose deprivation. Organotypic hippocampal slice cultures were prepared from 8 to 10-day-old rats and maintained for 10 to 12 days in vitro. Pre-treatment with Ac-YVAD-cmk prior to 45 min oxygen and glucose deprivation was neuroprotective as measured by propidium iodide uptake, with an EC(50) between 1 and 10 micromol/l. Ac-YVAD-cmk was also able to preserve synaptic function in the organotypic hippocampal slice cultures 24 h after oxygen and glucose deprivation. Ac-YVAD-cmk prevented the increase in histone-associated DNA fragmentation induced by oxygen and glucose deprivation. Interleukin-1beta did not reverse the protective effect of Ac-YVAD-cmk, and interleukin-1 receptor antagonist alone was not protective. These results show that caspase inhibitors are neuroprotective in a hippocampal slice culture system, using structural, biochemical and electrophysiological endpoints, and that this effect is not a result of inhibition of interleukin-1beta production. PMID- 10837799 TI - DNA fragmentation factor 45 deficient mice exhibit enhanced spatial learning and memory compared to wild-type control mice. AB - Programmed cell death or apoptosis is a highly regulated physiological process that is critical in development, particularly in the central nervous system. The DNA fragmentation factor 45 (DFF45 or ICAD) is a subunit of a heterodimeric DNase complex that is crucial for DNA fragmentation and normal apoptosis. To examine the neurobiological consequences of lacking DNA fragmentation and timely apoptosis during mouse development in vivo, we compared spatial learning behaviors in DFF45 mutant and wild-type control mice. We found that DFF45 mutant mice exhibit enhanced spatial learning and memory compared to wild-type mice. Moreover, both the granule cell density and total granule cell number in the hippocampal dentate gyrus region are higher in the DFF45 mutant brains than in the wild-type brains. We propose that the increase in granule cell number in the dentate region due to the DFF45 mutation changes the neuronal network underlying spatial learning and memory in DFF45 mutant mice. PMID- 10837800 TI - E-64-d prevents both calpain upregulation and apoptosis in the lesion and penumbra following spinal cord injury in rats. AB - Calpain, a Ca(2+)-dependent cysteine protease, has been implicated in cytoskeletal protein degradation and neurodegeneration in the lesion and adjacent areas following spinal cord injury (SCI). To attenuate apoptosis or programmed cell death (PCD) in SCI, we treated injured rats with E-64-d, a cell permeable and selective inhibitor of calpain. SCI was induced on T12 by the weight-drop (40 g-cm force) method. Within 15 min, E-64-d (1 mg/kg) in 1.5% DMSO was administered i.v. to the SCI rats. Following 24 h treatment, a 5-cm long spinal cord section with the lesion in the center was collected. The spinal cord section was divided equally into five 1-cm segments (S1: distant rostral, S2: near rostral, S3: lesion or injury, S4: near caudal and S5: distant caudal) for analysis. Determination of mRNA levels by reverse transcriptase-polymerase chain reaction (RT-PCR) indicated that ratios of bax/bcl-2 and calpain/calpastatin were increased in spinal cord segments from injured rats compared to controls. Degradation of the 68-kD neurofilament protein and internucleosomal DNA fragmentation were also increased. All of these changes were maximally increased in the lesion and gradually decreased in the adjacent areas of SCI rats, while largely undetectable in E-64-d treated rats and absent in sham controls. The results indicate that apoptosis in rat SCI appears to be associated with calpain activity which can be attenuated by the calpain inhibitor E-64-d. PMID- 10837801 TI - Ectopic noradrenergic hyperinnervation does not functionally compensate for neonatal forebrain acetylcholine lesion. AB - Adult rats who have undergone neonatal 192 IgG-saporin induced lesions of forebrain acetylcholine (ACH) neurons are normal on many behavioral tasks. In this study we determined whether ectopic hippocampal ingrowths, a documented consequence of these neonatal cholinergic lesions, functionally compensate for ACH denervation in these rats. Neonatal rats underwent systemic 6-hydroxydopamine (6-OHDA) injections on postnatal days (PND) 1-3 to prevent the ingrowths, and/or intraventricular 192 IgG-saporin injections on PND 7. The 192 IgG-saporin profoundly reduced basal forebrain p75 neurotrophin receptor (p75(NTR)) immunoreactive (IR) neurons. The 6-OHDA treatment abolished hippocampal and cortical dopamine-beta-hydroxylase (DBH) IR terminals, indicating the absence of normal norepinephrine (NE) innervation. Ectopic DBH IR and p75(NTR) IR varicosities which occurred in the hippocampus of 192 IgG-saporin treated rats were also eliminated by 6-OHDA treatment. Behavioral testing in adulthood indicated no effect of the treatments on the Morris water maze. 192 IgG-saporin treatment caused perseveration during delayed spatial alternation (DSA) and increased working but not reference memory errors on the radial arm maze (RAM). The 6-OHDA plus 192 IgG-saporin treated rats did not differ from the 192 IgG saporin only rats on any task. These results indicate that ectopic hippocampal NE ingrowths do not functionally compensate for neonatal ACH lesions. Neonatal forebrain ACH lesion impairs working memory on the RAM but the absence of an effect on DSA contraindicates a basic dysfunction of short term memory. Despite severe combined neonatal loss of forebrain ACH and NE innervation, behavior is remarkably intact. PMID- 10837802 TI - c-Fos gene expression pattern in the hypothalamus and the preoptic area of defeminized rats. AB - The object of the present study was to determine the c-fos gene expression pattern in the hypothalamus (HYP) and the preoptic area (POA) after estradiol and testosterone priming during the critical period of sexual differentiation of the rat brain. Three-day-old female rats were injected s.c. with a single dose of 17beta-estradiol (200 microg), testosterone enantate (200 microg) or vehicle (corn oil). HYP and POA were dissected 2 h, 24 h and 14 days after treatments and on the day of vaginal opening (VO). Other animals, previously treated as above, were acutely injected with 17beta-estradiol (5 microg) on the day of VO; HYP and POA were obtained 3 h later. Total RNA was extracted and processed for semiquantitative RT-PCR. We observed that c-fos gene expression was markedly increased in POA of the animals treated with estradiol or testosterone 2 h after treatments, while a non-significant increase in c-fos gene expression was observed in the HYP of these animals. We found a significant increase in c-fos expression in HYP and POA on the day of VO in both estradiol and testosterone defeminized rats. Interestingly, the acute estradiol administration on the day of VO did not induce c-fos gene expression in either HYP or POA of defeminized animals, instead a diminution in its expression was observed in animals treated with testosterone in POA. The overall results suggest that estradiol and testosterone imprinting during critical postnatal period of sexual differentiation of the brain permanently modifies the regulation of c-fos gene expression. PMID- 10837803 TI - The contribution of the median preoptic nucleus to renal sympathetic nerve activity increased by intracerebroventricular injection of hypertonic saline in the rat. AB - The median preoptic nucleus (MnPO) of the hypothalamus is involved in the osmotic control of neurohypophysial hormone release and drinking behavior. At the same time, renal sympathetic nerves exert multiple effects on renal functions such as regulating renal blood flow and urinary sodium excretion. We made the hypothesis that the MnPO may also regulate body fluid balance by exerting an influence on renal sympathetic nerve activity (RSNA). In this study we examined the effect of electrical stimulation of the MnPO on RSNA and the contribution of the MnPO to the change of RSNA induced by intracerebroventricular injection of hypertonic saline in the male Wistar rat. Electrical stimulation of the MnPO and the paraventricular nucleus of the hypothalamus (PVN) elicited an increase in RSNA. This increase of RSNA elicited by electrical stimulation of the MnPO was reduced by microinjections (100 nl) of 10% lidocaine or 4 mM cobaltous chloride (a synaptic transmission blocking agent) bilaterally into the PVN. Both RSNA and the mean arterial pressure (MAP) were increased by the injection of 1.5 M NaCl into the third ventricle, although heart rate (HR) was not significantly changed. These responses of RSNA and MAP were diminished by microinjection of 10% lidocaine (100 nl) into the MnPO. Our results suggest that the MnPO is involved in body fluid regulation not only by controlling vasopressin secretion and water intake but also by modulating central sympathetic outflow which regulates body fluid balance through an effect on the kidney. PMID- 10837804 TI - Cyclopentenone prostaglandin 15-deoxy-Delta(12,14)-prostaglandin J(2) acts as a general inhibitor of inflammatory responses in activated BV-2 microglial cells. AB - 15-deoxy-Delta(12,14)-PGJ(2), a cyclopentenone derivative of PGD(2), was recently reported [Petrova et al., Proc. Natl. Acad. Sci. USA 96 (1999) 4668-4673] to suppress inducible nitric oxide synthase (iNOS) production in microglia and mixed glial cultures stimulated with lipopolysaccharide (LPS). We report here that in addition to suppressing iNOS production, 15d-PGJ(2) also decreases the production of tumor necrosis factor alpha (TNFalpha), interleukin-1 beta (IL-1beta) and cyclooxygenase-2 (COX-2) in LPS-stimulated BV-2 microglial cells, thereby acting as a general inhibitor of microglial activation. Concomitantly, 15d-PGJ(2) itself up-regulates the production of the antioxidant enzyme heme oxygenase-1 (HO-1) and increases intracellular total glutathione levels. To test if increased HO-1 levels were involved in the ability of 15d-PGJ(2) to block microglial activation, we used a HO-1 inhibitor that could block the activity of HO-1. The presence of the HO-1 inhibitor did not alter the 15d-PGJ(2)-induced inhibition of LPS stimulated iNOS and TNFalpha protein levels, and led to only a partial reduction in the protection offered by 15d-PGJ(2) against LPS-induced nitrite production. These results suggest that HO-1 upregulation by 15d-PGJ(2) is not the primary pathway responsible for the anti-inflammatory action of 15d-PGJ(2) in microglial cells. PMID- 10837805 TI - DARPP-32 knockout mice exhibit impaired reversal learning in a discriminated operant task. AB - The current study was conducted to examine the performance of mice with a targeted deletion of the gene for DARPP-32 in a discriminated operant task using food reinforcement. DARPP-32 plays a central role in regulating the efficacy of dopaminergic neurotransmission. Initially, wild-type and DARPP-32 knockout mice were trained to nose-poke for food on a continuous reinforcement schedule. The minimum response requirement was increased every 5 days until the animals were responding on an FR-15 schedule of reinforcement. At the completion of extensive operant training, reversal learning was assessed. Wild-type and DARPP-32 knockout mice exhibited equivalent performance during acquisition of this task, with both groups increasing operant responding as the schedule of reinforcement was raised. However, significant differences in discrimination learning were observed during the reversal phase, with DARPP-32 knockout mice requiring significantly more trials to reach criterion than wild-type controls. These results provide evidence for a functional role of DARPP-32 in the mediation of processes underlying learning and memory. PMID- 10837806 TI - Localization of 5-ht(5A) receptor-like immunoreactivity in the rat brain. AB - 5-Hydroxytryptamine exerts modulatory physiological effects on both the central and peripheral nervous systems by activation of discrete receptor families. 5 ht(5A) is among the recently cloned, novel 5-HT receptors and currently under investigation to identify its pharmacological characteristics and potential physiological function(s). In this study, antibodies raised to a 5-ht(5A) specific peptide were characterized using dot blot, sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and immunohistochemistry, and the distribution of 5-ht(5A)-like immunoreactive material determined in rat brain. A major band of 41 kDa was observed following SDS-PAGE, corresponding to the predicted size of this receptor. 5-ht(5A)-like immunoreactivity was detected in areas known to have significant serotonergic input, such as hypothalamic and amygdaloid nuclei. Interestingly, 5-ht(5A)-like immunoreactivity showed a predilection for the suprachiasmatic nucleus, suggesting its possible role in the regulation of circadian rhythms, a function previously ascribed to 5-HT(1A) and 5 HT(7) receptors. PMID- 10837807 TI - Block of hippocampal CAN channels by flufenamate. AB - Ca(2+)-activated non-selective cation (CAN) channels are activated by cytoplasmic Ca(2+) and I(CAN) underlies many slow depolarizing processes in neurons including a putative role in excitotoxicity. CAN channels in many non-neuronal cells are blocked by non-steroidal antiinflammatory drugs that are derivatives of diphenylamine-2-carboxylate (DPC). The DPC derivative flufenamate (FFA) has a complex effect on certain neurons, whereby it blocks CAN channels and increases [Ca(2+)](i). We report here that FFA, but not the parent compound, DPC, blocks CAN channels in hippocampal CA1 neurons. As was the case in other neurons, the effects of FFA are complex and include a maintained rise in [Ca(2+)](i). Furthermore, the CAN channel blocking ability of FFA persists even when the channels have been potentiated by a Ca(2+)-dependent process. The use of a CAN channel-blocking drug is important for delineating CAN channel-dependent processes and may provide a basis for therapy for CAN channel-dependent events in ischemia. PMID- 10837808 TI - Effect of streptozotocin-induced diabetes on NGF, P75(NTR) and TrkA content of prevertebral and paravertebral rat sympathetic ganglia. AB - Diabetic autonomic neuropathy results in significant morbidity and mortality. Both diabetic humans and experimental animals show neuroaxonal dystrophy of autonomic nerve terminals, particularly in the prevertebral superior mesenteric ganglia (SMG) and celiac ganglia (CG) which innervate the hyperplastic/hypertrophic diabetic small intestine. Previously, investigators suggested that disturbances in ganglionic nerve growth factor (NGF) content or transport might play a pathogenetic role in diabetic autonomic pathology. To test this hypothesis, we measured NGF content and NGF receptor expression, p75(NTR) (low affinity neurotrophin receptor) and trkA (high affinity NGF receptor), in control and diabetic rat SMG, CG and superior cervical ganglia (SCG). Surprisingly, rather than a decrease, we observed an approximate doubling of NGF content in the diabetic SMG and CG, a result which reflects increased NGF content in the hyperplastic diabetic alimentary tract. No change in NGF content was detected in the diabetic SCG which is relatively spared in experimental diabetic autonomic neuropathy. NGF receptor expression was not consistently altered in any of the autonomic ganglia. These observations suggest that increased NGF content in sympathetic ganglia innervating the diabetic alimentary tract coupled with intact receptor expression may produce aberrant axonal sprouting and neuroaxonal dystrophy. PMID- 10837809 TI - Nicotine administration enhances NPY expression in the rat hypothalamus. AB - Epidemiological studies have shown an inverse relationship between cigarette smoking and body weight. In rodents, a negative correlation between nicotine and body weight has been reported, but this observation was largely derived from studies where relatively high doses of nicotine ( approximately 12 mg/kg/day) were used. In the current study, we showed that a negative relationship also holds for low doses of nicotine that are comparable to that consumed by average human smokers (<6 mg/kg/day). We also demonstrated that 14 days of nicotine administration (4 mg/kg/day) reduced average daily food intake by 19.5% (P<0.01) in the free-feeding nicotine-treated group compared to saline controls. No significant differences in body weight were detected between the nicotine-treated and pair-fed groups. To determine whether the effects of nicotine on food intake and body weight were related to neuropeptide Y (NPY) expression, semi quantitative reverse transcription-polymerase chain reaction (RT-PCR) and radioimmunoassay were utilized to measure NPY mRNA and peptide levels in various regions of the hypothalamus. Significantly higher levels of NPY mRNA (ca. 20-50%) and peptide (ca. 24-69%) were only detected in the nicotine-treated groups. In addition, significantly higher NPY contents were also obtained in two hypothalamic areas of pair-fed control animals. In summary, our data suggest that the pharmacological effects of nicotine on food intake and body weight may be mediated by changes in hypothalamic NPY levels, a neuropeptide that is pivotal to the hypothalamic regulation of food intake. PMID- 10837810 TI - Reducing conditions differentially affect the functional and structural properties of group-I and -II metabotropic glutamate receptors. AB - We have examined the influence of reducing conditions on the activity of group-I or -II metabotropic glutamate receptors. In cultured cerebellar granule cells or in hippocampal slices, the reducing agent dithiothreitol (DTT) inhibited the stimulation of polyphosphoinositide (PPI) hydrolysis elicited by group-I mGlu receptor agonists without affecting responses to norepinephrine or carbamylcholine. Similarly, DTT reduced the increase in intracellular free Ca(2+) induced by glutamate in HEK-293 cells expressing mGlu5 receptors. In adult hippocampal slices, the selective group-II mGlu receptor agonist, (2S,1'R,2'R,3'R)-2-(2, 3-dicarboxycyclopropyl)glycine (DCG-IV) had no effect per se on PPI hydrolysis, but potentiated the response to quisqualate. Although DTT substantially attenuated the action of quisqualate, it did not affect the potentiation by DCG-IV, suggesting that group-II mGlu receptors are resistant to extracellular reduction. Accordingly, DTT did not affect the inhibition of forskolin-stimulated cAMP formation induced by maximally effective concentrations of group-II mGlu receptor agonists in hippocampal slices or in CHO cells expressing mGlu2 receptors. At structural level, DTT differentially affected the aggregation state of mGlu1a, -2/3 or -5 receptors. In immunoblots performed under non-reducing conditions, mGlu1a, -2/3 or -5 antibodies labeled exclusively a high molecular weight band, corresponding to receptor dimers. Under reducing conditions, mGlu1a or -5 receptors were detected as monomers, whereas a large proportion of mGlu2/3 receptors was still present in a dimeric form. We conclude that reducing conditions differentially influence the aggregation state of group I and -II mGlu receptors and suggest that dimerization affects the functional activity of native mGlu receptors. PMID- 10837811 TI - The effects of thrombin preconditioning on focal cerebral ischemia in rats. AB - Our previous studies have shown that prior intracerebral infusion of a low dose of thrombin (thrombin preconditioning; TPC) reduces the brain edema that follows a subsequent intracerebral infusion of a high dose of thrombin or an intracerebral hemorrhage. In vitro studies have also demonstrated that low concentrations of thrombin protect neurons and astrocytes from hypoglycemia and oxidative stress-induced damage. This study, therefore, examines the hypothesis that TPC would offer protection from ischemic brain damage in vivo. This was a blinded design study. The rat brain was preconditioned with 1 U thrombin by direct infusion into the left caudate nucleus. Seven days after thrombin pretreatment, permanent middle cerebral artery occlusion (MCAO) was induced. Twenty-four hours post-ischemia, neurological deficit was evaluated and infarction volume, brain water and ion contents were measured. Compared to saline treated rats, thrombin pretreatment significantly attenuated brain infarction in cortex (90+/-33 vs. 273+/-22 mm(3); P<0.05) and basal ganglia (56+/-17 vs. 119+/ 12 mm(3); P<0.05) that followed 24 h of permanent MCAO. TPC also reduced the brain edema in cortex and basal ganglia by 50 and 53% (P<0.05). Neurological deficit was improved in thrombin pretreatment group (P<0.05). These effects of TPC were, in part, prevented by co-injection of hirudin, a thrombin inhibitor, indicating that the protection was indeed thrombin mediated. Cerebral TPC significantly reduces ischemic brain damage, perhaps by activation of the thrombin receptor. This finding provides a new mechanism by which to study ischemic tolerance. PMID- 10837812 TI - Correlations of norepinephrine release in the paraventricular nucleus with plasma corticosterone and leptin after systemic lipopolysaccharide: blockade by soluble IL-1 receptor. AB - The purpose of the study was to investigate the effects of systemic lipopolysaccharide (LPS) on norepinephrine (NE) release in the paraventricular nucleus (PVN) and on plasma concentrations of corticosterone and leptin. Soluble IL-1 receptor (sIL-1R) was used to determine the role of interleukin-1 (IL-1) in these effects. Adult male rats were implanted with a push-pull cannula in the PVN and a jugular catheter to facilitate blood sampling. On the day of the experiment, after the collection of a pretreatment blood and perfusate sample, rats were injected (i.p.) with the vehicle for LPS (saline), 2.5 or 10 microg/kg BW LPS. Other groups of animals were treated i.p. with 25 microg of sIL-1R, or a combination of 10 microg/kg BW of LPS and 25 microg of sIL-1R, 5 min before and 90 min after LPS. Blood and perfusate samples were collected at 30-min intervals for 6 h. NE concentrations in the perfusate were measured using HPLC-EC and corticosterone and leptin levels in the plasma were measured using radioimmunoassay. NE release in the PVN was dose dependent and increased significantly within 90 min in response to the high dose of LPS and reached maximum levels around 180 min before declining gradually to pretreatment levels at 330 min. The corticosterone profile in LPS-treated animals was similar to the NE release profile in the PVN. In contrast, the LPS-induced increase in leptin levels reached a maximum at 210 min and remained elevated even at the end of the observation period. Treatment with sIL-1R completely blocked the LPS-induced effects. It is concluded that LPS stimulates NE release in the PVN and increases plasma concentrations of corticosterone and leptin and that these effects are mediated at least in part by IL-1. PMID- 10837813 TI - Octopamine is the synaptic transmitter between identified neurons in the buccal feeding network of the pond snail lymnaea stagnalis. AB - We report the pharmacological properties of synaptic connections from the three octopamine-containing OC interneurons to identified buccal feeding neurons in the pond snail, Lymnaea stagnalis. Intracellular stimulation of an OC interneuron evokes inhibitory postsynaptic potentials in the B3 motoneurons and N2 (d) interneurons, while the synapse between OC and N3 (phasic) interneurons has two components: an initial electrical excitation followed by chemical inhibition. All synaptic responses persist in a saline with elevated calcium and magnesium suggesting that the connections are monosynaptic. Local perfusion of 10(-4) M octopamine produces the same inhibitory membrane responses from these buccal neurons as OC stimulation. These responses also persist in high Mg(2+)/Ca(2+) saline indicating direct membrane effects. The similarities in reversal potentials for the synaptic hyperpolarization evoked on B3 neurons after OC stimulation (-89.0 mV, S.E.M.=14.1, n=10) and the octopamine response of the B3 neurons (-84.7 mV, S.E.M.=6.6, n=6) indicate that increased K(+)-conductance underlies both responses. Bath application of the octopaminergic drugs phentolamine (10(-6) M), epinastine (10(-6) M) or DCDM (10(-4) M) blocks the inhibitory synapse onto B3 or N2 neurons and the chemical component of the N3 response. They also block the octopamine-evoked inhibition of B3, N2 and N3 neurons. NC-7 (2x10(-5) M) has a hyperpolarizing agonist effect (like octopamine) on these neurons and also blocks their chemical synaptic input from the OC interneurons. These results provide pharmacological evidence that the neurotransmitter between the octopamine-immunopositive OC interneurons and its followers is octopamine. This is the first example of identified octopaminergic synaptic connections within the snail CNS. PMID- 10837814 TI - Medullary and intrathecal injections of 17beta-estradiol in male rats. AB - The following experiments were designed to investigate the role of estrogen in central autonomic nuclei on autonomic tone and reflex control of heart rate. Male Sprague-Dawley rats were anesthetized with sodium thiobutabarbital (100 mg/kg) and instrumented to record blood pressure and heart rate. Efferent vagal and renal nerve activities were recorded and used to assess changes in parasympathetic and sympathetic tone, respectively. The cardiac baroreflex was evoked using a single bolus injection of phenylephrine (0.1 mg/kg) both before and following either intrathecal injection of estrogen (0.5 microM; 1 microl) to influence sympathetic preganglionic neurons of the intermediolateral cell column or bilateral injection of estrogen (0.5 microM; 100 nl/side) into the nucleus tractus solitarius, rostral ventrolateral medulla or nucleus ambiguus. The cardiac baroreflex was significantly enhanced following both intrathecal and medullary injections of estrogen. Efferent vagal nerve activity was significantly increased following injection of estrogen into the nucleus tractus solitarius, nucleus ambiguus and the intrathecal space. Renal sympathetic nerve activity was significantly depressed following injection of estrogen into the nucleus tractus solitarius, rostral ventrolateral medulla and the intrathecal space. In all cases, simultaneous injection of estrogen with the selective estrogen receptor antagonist, ICI 182,780 (1 pM) blocked all previously observed changes in baroreflex function and autonomic tone. These results demonstrate a role for estrogen in the reflex control of heart rate and as a central modulator of autonomic tone in male rats. PMID- 10837815 TI - Involvement of the noradrenergic system in modulating the blink reflex in humans. AB - Several researches have shown that the spinal reflex transmission in animals, as well as humans, was inhibited by alpha(2)-agonists, due to a disfacilitation of tonic noradrenergic control on motoneuronal output. To understand better the mechanisms regulating certain aspects of motor activity, here we reinvestigated the possible role of noradrenergic systems in modulating reflex activity of the brainstem in humans. To this aim, blink reflex responses (R1 and R2) evoked by electrical stimulation of the supraorbital nerve were electromyographically recorded in healthy volunteers. Both R1 and R2 areas were measured at 10-min intervals before and after i.v. injection of alpha(2)-agonist clonidine (0.5 microg/kg). The substance induced consistent depression of R1, which reached its maximum 40 min after drug administration (-43% of the control values). Ipsilateral R2 area resulted little affected by clonidine (-15% at 50 min), whereas no effects were observed in contralateral R2. Blood pressure values were never altered by drug injections. These results, taken together with previous observations, support the hypothesis that alpha(2)-agonist substances may cause a transient inactivation of noradrenergic neurons, thus releasing neurons involved in the circuitry of the blink reflex from a facilitatory drive. Since clonidine differentially modulated blink reflex responses, it is likely to assume that such a disfacilitation concerns mostly pontine units mediating the R1. However, the complexity of clonidine's effects at multiple pre- and postsynaptic sites does not allow us to exclude that other systems are involved in the alpha(2)-mediated control of facial motoneurons. PMID- 10837816 TI - Brain multiparametric responses to carbon monoxide exposure in the aging rat. AB - The multiparametric monitoring system was applied to study the effects of 2000 ppm carbon monoxide (CO) on brain functions in vivo in the aging rat. The vasodilatory (non hypoxic) effects of CO on CBF in normal adult rats, which were shown in concentrations of 1000-2000 ppm involved the effect of nitric oxide (NO). Energy metabolism was evaluated by optical monitoring of CBF and mitochondrial function by fluorometry of NADH. Ionic homeostasis was evaluated by monitoring the extracellular level of K(+) and H(+) and the DC steady potential. Seven aging rats (24 months) were exposed to 2000 ppm for 60 min and 120 min of recovery, while five control rats were exposed to air under the same conditions. A comparison between the CO group and the control group showed that the changes in CBF, NADH and light reflectance were not statistically significant while extracellular K(+) was elevated and tissue pH became more acidic. Thus, the typical CO induced increase in CBF, was not recorded in the aging rats. We concluded that the brain vasodilatory response to CO was not active in the aging rat, while the ionic homeostasis responses were similar to those found in the adult rat. PMID- 10837817 TI - Ionic permeability of the opossum sciatic nerve perineurium, examined using electrophysiological and electron microscopic techniques. AB - A parallel electrophysiological and electron microscopic study was used to assess the ionic permeability of the sciatic nerve perineurium of the opossum Monodelphis domestica. The electrophysiological method was used to monitor permeability to K(+), followed by combined electron microscopy and X-ray probe analysis to monitor permeability to the electron-dense tracer lanthanum. Isolated but intact nerves were mounted in a 'grease gap' chamber for extracellular measurement of DC potential and compound action potential (CAP). Challenge with 100 mM [K(+)] Ringer was used to assess the K(+) permeability of the perineurium, since a change in DC potential (DeltaDC) under these conditions reflected changes in the axonal resting membrane potential. There was no detectable change in DC potential or CAP to the first K(+) challenge (n=71 nerves) indicating negligible K(+) permeability under control conditions. The inflammatory mediators histamine 0.1-40 mg/ml (1. 3-130 mM), bradykinin (0.1-4.7 mM) and 5HT (serotonin) 0.1-5.0 mg/ml (0.5-23.5 mM) caused no measurable DeltaDC on subsequent challenge with 100 mM [K(+)] Ringer, indicating no effect on perineurial K(+) permeability. In nerves exposed to the bile salt sodium deoxycholate (DOC, 6 min, 4 mM), challenge with elevated K(+) Ringer caused a dose-dependent DeltaDC in the range 10-100 mM [K(+)] (1.67+/-0.17 mV in 100 mM [K(+)], n=20), indicating increased perineurial permeability caused by DOC, but the response was smaller than that previously reported for the frog perineurium. Lanthanum was observed in the outer layers of the perineurium, but was not seen to penetrate the endoneurium in any of the nerves examined (n=51), even after DOC application. This study shows that the combined electrophysiological and electron microscopic technique for monitoring ionic permeability can be applied to mammalian nerve, and suggests that the opossum perineurium is more resistant to tight junction opening by chemical modulators than is the frog perineurium. PMID- 10837818 TI - Laser scanning and electron microscopic evidence for rapid and specific in vivo labelling of cholinergic neurons in the rat basal forebrain with fluorochromated antibodies. AB - Recently developed methods for the selective labelling of cholinergic basal forebrain neurons containing the low-affinity neurotrophin receptor p75 (p75(NTR)) in vivo and in vitro are based on carbocyanine 3 (Cy3)-tagged antibodies directed against p75(NTR). The present study focuses on the maintenance of this neuronal label after injection of such fluorescent antibodies into the cerebral ventricle. One, 3, and 10 days after injection this marker exclusively stains neurons immunoreactive for the cholinergic markers choline acetyltransferase and vesicular acetylcholine transporter in the rat medial septum, diagonal band and nucleus basalis. Thirty days after injection the in vivo labelling was nearly abolished. Predominant labelling of lysosomes was shown by electron microscopic analysis following photoconversion of the Cy3-label to an electron-dense reaction product. The pre-labelling of cholinergic neurons might facilitate pharmacological and electrophysiological approaches in living slices and cell culture systems as well as detailed investigations focused on the transport of neurotrophins in vivo and in animals with experimentally altered p75(NTR) expression. PMID- 10837820 TI - Effects of REM sleep deprivation on cholinergic receptor sensitivity and passive avoidance behavior in clomipramine model of depression. AB - This study investigated the effects of REM (rapid eye movement) sleep deprivation (RSD) on the activity of central cholinergic receptors and passive avoidance retention in rats treated neonatally with clomipramine. Male rat pups treated with clomipramine (15 mg/kg, s.c.) twice daily from postnatal day 5 to 21 were subjected to RSD procedure at three months of age, for 4 days consecutively. In the post-RSD phase, RSD-control rats showed a significantly enhanced cholinomimetic-induced hypothermia and an improved retention in passive avoidance task. However, these measures were not significantly different in RSD experimental group as compared to rats treated neonatally with saline. These results suggest that RSD reverses the sensitivity of central cholinergic receptors in rats given clomipramine neonatally, and this mechanism may be involved in mediating the antidepressant effects of RSD treatment in clomipramine model of depression. PMID- 10837819 TI - The dopamine D3 receptor antagonist nafadotride inhibits development of locomotor sensitization to amphetamine. AB - Behavioral sensitization is a well-studied model of behavioral plasticity mediated at least in part by dopaminergic systems believed to play an important role in several psychiatric conditions. In the rodent, locomotion is regulated by the opposing balance of D3 and D2 receptors, with D2 activation increasing and D3 stimulation inhibiting locomotion. However, receptor occupancy of D3 dopamine receptors is far greater than D2 or D1 occupancy at typical post-stimulant dopamine concentrations. We therefore hypothesized that tolerance of D3 receptor inhibition of locomotion contributes to the development of sensitization. To test this hypothesis, we examined the effect of the D3 receptor antagonist nafadotride on sensitization. As predicted, nafadotride inhibits augmentation of the locomotion response to repetitive amphetamine. This finding is consistent with the proposed model of adaptive down-regulation of D3 dopamine receptor function contributing to the development of behavioral sensitization. PMID- 10837821 TI - Differential modulatory roles of cholera toxin and pertussis toxin in the regulation of pain responses induced by excitatory amino acids administered intrathecally in mice. AB - The present study was designed to characterize the possible roles of spinally located cholera toxin (CTX)- and pertussis toxin (PTX)-sensitive G-proteins in excitatory amino acids induced pain response. Intrathecal (i.t.) injection of glutamate (20 microg), N-methyl-D-aspartic acid (NMDA; 60 ng), alpha-amino-3 hydroxy-5-methylisoxazole-4-propionic acid (AMPA; 13 ng), and kainic acid (12 ng) showed pain response. Pretreatment with CTX (0.05 and 0.5 microg, i.t.) attenuated pain response induced by glutamate, NMDA, AMPA and kainic acid administered i.t. in a dose-dependent manner. On the other hand, i.t. pretreatment with PTX further increased the pain response induced by glutamate, NMDA, AMPA and kainic acid administered i.t., especially at the dose of 0.5 microg. Our results suggest that, at the spinal cord level, CTX- and PTX sensitive G-proteins appear to play opposite roles in modulating the pain response induced by spinally administered. Furthermore, CTX- and PTX-sensitive G proteins appear to modulate pain response induced by stimuli of both NMDA and non NMDA glutamate receptors. PMID- 10837822 TI - D1 dopamine receptor activation reduces extracellular glutamate and GABA concentrations in the medial prefrontal cortex. AB - The present study examined effect of administration of a selective D1 dopamine receptor agonist, SKF38393 on extracellular concentrations of glutamate (Glu) and gamma-aminobutyric acid (GABA) in mPFC, by using in vivo microdialysis. Perfusion with SKF38393 via a dialysis probe reduced concentrations of both Glu and GABA dose-relatedly, and these effects were prevented by co-perfusion with a D1 dopamine receptor antagonist, SCH23390 (40 microM). These results suggested that the dopaminergic hyperactivity may lead to the hypofunction of glutamatergic and GABAergic systems in mPFC via D1 dopamine receptor stimulation. PMID- 10837823 TI - C-nociceptor sensitization by isoprostanes is cyclooxygenase dependent. AB - Isoprostane E(2) (8-iso-PGE) and F(2alpha) (8-iso-PGF) sensitize nociceptors and capsaicin-sensitive DRG neurons. In this study we investigated the cyclooxygenase dependence of isoprostane-induced C-nociceptor sensitization. Systemic pretreatment of rats with ketorolac (1 and 10 mg/kg) abolished 8-iso-PGF sensitization and reduced the effects of 8-iso-PGE. Ibuprofen (30 mg/kg) blocked all sensitizing effects. These data suggest that some algesic properties of isoprostanes are mediated via prostanoid synthesis. PMID- 10837824 TI - Kindling induces a transient suppression of afterhyperpolarization in rat subicular neurons. AB - To determine whether chronic epilepsy induces persistent cellular changes in subicular neurons intracellular recordings were used to compare membrane properties of control and kindled rats. In both, control and kindled preparations the subiculum contained regular firing cells and an extensive sub-population of bursting cells expressing amplifying membrane characteristics. Subicular cells showed a transient depression of the fast and slow AHP in the course of kindling that may contribute to the induction but not permanence of the kindled state. PMID- 10837825 TI - Similar poly(C)-sensitive RNA-binding complexes regulate the stability of the heavy and light neurofilament mRNAs. AB - The potential role of RNA processing in regulating neurofilament (NF) subunit expression and in mediating the neuropathic effects of NF transgenes was explored by determining whether similar regulatory elements and cognate binding factors are present in NF mRNAs. Gel-shift studies were used to compare RNA-binding complexes that assemble on the 3'UTR of the heavy (NF-H), mid-sized (NF-M) and light (NF-L) NF mRNAs when radioactive RNA probes are incubated with high-speed supernatants (S100) of rat brain homogenates. RNA-binding complexes were characterized by their rate of migration in non-denaturing gels and by their ability to be competed with specific homoribopolymers. Similar RNA-binding complexes formed on probes to the 3'UTRs of NF-L and NF-H mRNAs. The complexes were competed with poly(C) and are referred to as poly(C)-sensitive complexes. Their binding sites were localized to a 36 nt sequence in the mid-distal region of the NF-H 3'UTR and to a 45 nt sequence at the proximal edge of the 3'UTR of the NF-L transcript. Although the binding sites showed limited sequence homology, the complexes were cross-competed with unlabeled probes and radioactivity in either probe was cross-linked to a 43 kDa protein. The 43 kDa protein also bound directly to NF-L and NF-H probes in Northwestern blots. Functional studies showed that deletion of the binding sites markedly increased expression of a luciferase reporter gene containing the 3'UTR of NF-L or NF-H by stabilizing the fusion transcripts. Point mutations in the NF-H binding site which prevented formation of the poly(C)-sensitive complex also stabilized the fusion mRNA. The findings reveal a common destabilizing element in the 3'UTR of NF-L and NF-H mRNAs that may be important in coordinating NF subunit expression and in mediating the neuropathic effects of the NF-L and NF-H transgenes in transgenic mice. PMID- 10837826 TI - S100 proteins in Corpora amylacea from normal human brain. AB - Corpora amylacea (C.A.) also named polyglucosan bodies (P.B.) are one of the hallmarks of normal brain aging. Although their functions are not yet clear, C.A. increase in number in patients suffering from neurodegenerative diseases. C.A. contain 88% of hexoses and 4% of proteins. Most of the proteins in C.A. are aging or stress proteins such as heat shock proteins, ubiquitinated proteins and advanced glycation end products which are also proinflammatory products. Stimulated by the potential role played by some S100 proteins in the inflammatory process which may be triggered in C.A., we investigated, by immunohistochemistry, the presence of different S100 proteins (S100A1, S100A2, S100A3, S100A4, S100A5, S100A6, S100A8, S100A9, S100A12 and S100B) in C.A. from normal human brain. Among the ten S100 proteins analyzed, nine (S100A) were detected in C.A. Three S100 proteins (S100A8, S100A9, S100A12) which are highly expressed in activated macrophages and used as inflammatory markers were detected in C.A. S100A8 was, in addition, found in thick neuronal processes from the pons. One (S100B) could not be found in C.A. although it was highly expressed in astrocytes. In C.A., the staining intensity was estimated by computer-assisted microscopy and gave the following order: S100A1 congruent withS100A8 congruent with S100A9>S100A5> or =S100A4>S100A12>S100A6> S100A2=S100A3. The potential inflammatory role played by S100 proteins in C.A. is discussed. PMID- 10837827 TI - Center surround receptive field structure of cone bipolar cells in primate retina. AB - In non-mammalian vertebrates, retinal bipolar cells show center-surround receptive field organization. In mammals, recordings from bipolar cells are rare and have not revealed a clear surround. Here we report center-surround receptive fields of identified cone bipolar cells in the macaque monkey retina. In the peripheral retina, cone bipolar cell nuclei were labeled in vitro with diamidino phenylindole (DAPI), targeted for recording under microscopic control, and anatomically identified by intracellular staining. Identified cells included 'diffuse' bipolar cells, which contact multiple cones, and 'midget' bipolar cells, which contact a single cone. Responses to flickering spots and annuli revealed a clear surround: both hyperpolarizing (OFF) and depolarizing (ON) cells responded with reversed polarity to annular stimuli. Center and surround dimensions were calculated for 12 bipolar cells from the spatial frequency response to drifting, sinusoidal luminance modulated gratings. The frequency response was bandpass and well fit by a difference of Gaussians receptive field model. Center diameters were all two to three times larger than known dendritic tree diameters for both diffuse and midget bipolar cells in the retinal periphery. In one instance intracellular staining revealed tracer spread between a recorded cell and its nearest neighbors, suggesting that homotypic electrical coupling may contribute to receptive field center size. Surrounds were around ten times larger in diameter than centers and in most cases the ratio of center to surround strength was approximately 1. We suggest that the center-surround receptive fields of the major primate ganglion cell types are established at the bipolar cell, probably by the circuitry of the outer retina. PMID- 10837828 TI - Time course of chromatic adaptation for color appearance and discrimination. AB - Adaptation to a steady background has a profound effect on both color appearance and discrimination. We determined the temporal characteristics of chromatic adaptation for appearance and discrimination along different color directions. Subjects were adapted to a large uniform background made up of a CRT screen and a 45x64 degrees wall, illuminated by computer controlled lamps. After an instant change in background color along a red-green or blue-yellow color axis, we measured thresholds for the detection of increments along the same axes at fixed times between 25 ms and 121 s. Analogously, color appearance was determined using achromatic matching. Three components of adaptation could be identified by their temporal characteristics. A slow exponential time course of adaptation with a half-life of about 20 s was common to appearance and discrimination. A faster component with a half-life of 40-70 ms--probably due to photoreceptor adaptation- was also common to both. Exclusive for color appearance, there was a third, extremely rapid mechanism with a half-life faster than 10 ms. This instantaneous process explained more than 50% of total adaptation for color appearance and could be shown to act in a multiplicative manner. We conclude that this instantaneous adaptation mechanism for color appearance is situated at a later processing stage, after mechanisms common to appearance and discrimination, and is based on multiplicative spatial interactions rather than on local, temporal adaptational processes. Color appearance, and thus color constancy, seems to be determined in large part by cortical computations. PMID- 10837829 TI - Effects of roll and pitch components in retinal flow on heading judgement. AB - We investigated effects of roll (rotation around line of sight) and pitch (rotation around the horizontal axis) components of retinal flow on heading judgement from visual motion information. It was found that performance level of human observers for yaw (rotation around the vertical axis) plus pitch is little different from that for only yaw although there is bias in perceived heading toward the fixation point, and that heading judgement is fairly robust with respect to roll. It was also found that there are some observers who can perceive heading with pitch, yaw and roll at a roll rate of 11.5 degrees /s without extra retinal information. It suggests that there exist compensation mechanisms for roll in the human visual system. PMID- 10837830 TI - Comparison of thresholds for high-speed drifting vernier and a matched temporal phase-discrimination task. AB - For rapidly translating targets, vernier thresholds correspond to millisecond asynchronies between targets. The 'temporal hypothesis' is that these thresholds reflect the limiting sensitivity of asynchrony detectors. Previous studies showed that temporal thresholds are generally higher than vernier thresholds, but failed to reject the 'temporal hypothesis' because stimuli had differing spatiotemporal characteristics, and temporal thresholds depend strongly on stimulus and task. Here we use matched grating stimuli to test - and reject - the temporal hypothesis. Expressed as asynchrony, temporal phase discrimination was typically 10-fold poorer than vernier thresholds, and differed in dependence on spatial frequency, temporal frequency, contrast, and susceptibility to stroboscopic masks. PMID- 10837831 TI - Unequal retinal and extra-retinal motion signals produce different perceived slants of moving surfaces. AB - Eye movements introduce retinal motion to the image and so affect motion cues to depth. For instance, the slant of a plane moving at right-angles to the observer is specified by translation and a component of relative motion such as shear. To a close approximation, the translation disappears from the image when the eye tracks the surface accurately with a pursuit eye movement. However, both translation and relative-motion components are needed to estimate slant accurately and unambiguously. During pursuit, therefore, an extra-retinal estimate of translation must be used by the observer to estimate surface slant. Extra-retinal and retinal estimates of translation speed are known to differ: a classic Aubert-Fleischl phenomenon was found for our stimuli. The decrease in perceived speed during pursuit predicts a corresponding increase in perceived slant when the eye tracks the surface. This was confirmed by comparing perceived slant in pursuit and eye-stationary conditions using slant-matching and slant estimation techniques. Moreover, the increase in perceived slant could be quantified solely on the basis of the perceived-speed data. We found no evidence that relative-motion estimates change between the two eye-movement conditions. A final experiment showed that perceived slant decreases when a fixed retinal shear is viewed with increasing pursuit speed, as predicted by the model. The implication of the results for recovering metric depth estimates from motion based cues is discussed. PMID- 10837832 TI - Temporal dependencies in resolving monocular and binocular cue conflict in slant perception. AB - Observers viewed large dichoptic patterns undergoing smooth temporal modulations or step changes in simulated slant or inclination under various conditions of disparity-perspective cue conflict and concordance. After presentation of each test surface, subjects adjusted a comparison surface to match the perceived slant or inclination of the test surface. Addition of conflicting perspective to disparity affected slant and inclination perception more for brief than for long presentations. Perspective had more influence for smooth temporal changes than for step changes in slant or inclination and for surfaces presented in isolation rather than with a zero disparity frame. These results indicate that conflicting perspective information plays a dominant role in determining the temporal properties of perceived slant and inclination. PMID- 10837833 TI - Development of chromatic and luminance detection contours using the sweep VEP. AB - This study measured the development of contrast-sweep VEP thresholds to a range of chromatic and luminance stimuli. Subjects were 14-32 week-old infants (n=21) and three adults. Stimuli were 1 c/d sine gratings reversed at 5.6 Hz. Chromaticity was varied from the L-M axis to an achromatic axis. VEP thresholds when plotted in L- and M-cone contrasts showed that: (1) VEP thresholds did not consistently locate the psychophysical isoluminance match under the same stimulus conditions; (2) About 50% of the data were described by independent chromatic and luminance mechanism, however, thresholds were limited by the cone contrast of the stimulus, phase cancellation between visual mechanisms, and the proper sampling of thresholds in L- and M-cone contrast space and; (3) No significant changes occurred in VEP detection contours across ages but suprathreshold VEP amplitudes suggested complex developmental changes. Data from previous studies showed similar results. PMID- 10837834 TI - Increment and decrement detection on temporally modulated fields. AB - Increment and decrement probe thresholds were measured during the presentation of two types of temporal masking stimuli. In Experiment 1, we measured thresholds for increment or decrement rectangular probes presented during the presentation of an increment or decrement Gaussian masking stimulus. We find that thresholds are higher when the probe and the Gaussian mask are of the same sign (e. g. both increments). However, both types of Gaussian mask raised increment and decrement probe thresholds above steady state conditions. In Experiment 2, we presented increment or decrement probes at one of eight possible phases of a 1 Hz luminance modulated sine wave. For both increment and decrement probes, threshold variation with phase is non-sinusoidal in shape, but increment and decrement probe thresholds vary as a function of the sinusoid phase. These experiments show that increment and decrement thresholds vary as a function of the adaptation state of the visual system, and as a function of the direction of change in the adaptation state. Data from both experiments are discussed in terms of a recent neurophysiological model [Hood & Graham (1998). Threshold fluctuations on temporally modulated backgrounds: a possible physiological explanation based upon a recent computational model. Visual Neuroscience, 15 (5), 957-967]. We find that the predicted ON- and OFF-pathway responses do not correlate in a straightforward manner with the psychophysical thresholds, suggesting that detection of increment and decrement probes may not be performed exclusively by one pathway. Our data have implications for modeling visual performance under conditions where visual adaptation is dynamic, such as when scanning complex images or natural scenes. PMID- 10837835 TI - The effect of contrast on reading speed in dyslexia. AB - Contrast coding has been reported to differ between dyslexic and normal readers. Dyslexic readers require higher levels of contrast to detect sinewave gratings for certain spatiotemporal conditions, and dyslexic readers show faster visual search at low contrast. We investigated whether these differences in early contrast coding generalize to reading performance by measuring reading speed as a function of text contrast for dyslexic children and adults and for age-matched controls. Contrast affected reading performance of dyslexic and normal readers similarly. For both groups, reading speed was relatively constant between 100 and 2% contrast, and decreased rapidly below 2% contrast. This pattern of results held true for both children and adults, for text with and without sentence context, across a range of character sizes, and for reading aloud and reading silently. We conclude that earlier findings of group differences in contrast effects on grating detection or visual search tasks do not generalize to reading. PMID- 10837836 TI - A model of the Parkinsonian visual system: support for the dark adaptation hypothesis. AB - Considerable evidence suggests that some visual abnormalities in Parkinson's disease are mediated by disruption of dopaminergic processes in the retina. Since dopamine is thought to be involved in the process of dark adaptation, and some of these abnormalities are similar to the changes which accompany dark adaptation in normal subjects, it has been proposed that the parkinsonian retina behaves as though inappropriately dark-adapted. In Parkinson's disease, the apparent contrast of peripherally viewed medium and high spatial frequency gratings is reduced. In our first experiment, normal subjects were dark-adapted, and were required to match the apparent contrast of a peripherally viewed grating to that of a foveally viewed grating. The results showed an interaction between spatial frequency and dark adaptation, reflecting a greater reduction in the apparent contrast of peripheral high spatial frequency gratings. In a second experiment, no effect of dark adaptation was found on the apparent spatial frequency of a peripherally viewed grating required to match that of a foveally viewed grating. The first experiment supports the dark adaptation hypothesis of parkinsonian vision, and the second suggests that the changes in apparent contrast are mediated by different amounts of change in contrast gain in central and peripheral vision, rather than by differential changes in receptive field size. PMID- 10837837 TI - Editorial PMID- 10837838 TI - Food craving and food "addiction": a critical review of the evidence from a biopsychosocial perspective. AB - Although certain commonalities exist between eating and drug use (mood effects, external cue-control of appetites, reinforcement, etc. ), it is argued that the vast majority of cases of (self-reported) food craving and food "addiction" should not be viewed as addictive behavior. An explanation is proposed that instead gives a prominent role to the psychological processes of ambivalence and attribution, operating together with normal mechanisms of appetite control, the hedonic effects of certain foods, and socially and culturally determined perceptions of appropriate intakes and uses of those foods. Ambivalence (e.g., "nice but naughty") about foods such as chocolate arises from the attitude that it is highly palatable but should be eaten with restraint. Attempts to restrict intake, however, cause the desire for chocolate to become more salient, an experience that is then labelled as a craving. This, together with a need to provide a reason for why resisting eating chocolate is difficult and sometimes fails, can, in turn, lead the individual to an explanation in terms of addiction (e.g., "chocoholism"). Moreishness ("causing a desire for more") occurs during, rather than preceding, an eating episode, and is experienced when the eater attempts to limit consumption before appetite for the food has been sated. PMID- 10837839 TI - Reversal of caffeine withdrawal by ingestion of a soft beverage. AB - Followlng regular use, acute cessation of caffeine is associated with a characteristic withdrawal syndrome. Despite this, caffeine remains popular with its consumers. The aim of this study was to examine the physiologic and psychologic effects of small caffeine doses, administered in the form of a market leading soft drink, on healthy women who were acutely withdrawn from caffeine. After 48-h abstinence and overnight fast, 11 healthy (22 to 40 years) female volunteers, all regular caffeine users (daily consumption 143 to 773 mg) consumed using a double-blind. randomized, controlled cross-over design either 2 tins of regular or caffeine-free Diet Coke. On both visits a Mars bar was eaten to prevent hypoglycaemia. Thus, the caffeine load was 76 or 10 mg respectively. Following ingestion of regular Diet Coke, there was a l0% fall in middle cerebral artery velocity (95% CI [6%-l4%], p < 0.005 versus caffeine free) and improvement in feelings of pleasure (p < 0.046) and energy (p < 0.037). Intellectual function (4-choice reaction time) was unaffected by caffeine status. On both visits, ingestion of Diet Coke induced a pressor response (maximum rise in systolic pressure +15+/- 2 mm Hg with caffeine and +l2 +/- 2 mm Hg with caffeine-free beverage, both p < 0.001 compared with baseline). In conclusion, in women acutely withdrawn from caffeine, ingestion of a popular soft beverage containing modest amounts of caffeine is associated with demonstrable physiologic and psychologic effects. PMID- 10837840 TI - The acute physiological and mood effects of tea and coffee: the role of caffeine level. AB - The objective of this study was to determine the effect of caffeine level in tea and coffee on acute physiological responses and mood. Randomised full crossover design in subjects after overnight caffeine abstention was studied. In study 1 (n = 17) the caffeine level was manipulated naturalistically by preparing tea and coffee at different strengths (1 or 2 cups equivalent). Caffeine levels were 37.5 and 75 mg in tea, 75 and 150 mg in coffee, with water and no-drink controls. In study 2 (n = 15) caffeine level alone was manipulated (water, decaffeinated tea, plus 0, 25, 50, 100, and 200 mg caffeine). Beverage volume and temperature (55 degrees C) were constant. SBP, DBP, heart rate, skin temperature, skin conductance, and mood were monitored over each 3-h study session. In study 1, tea and coffee produced mild autonomic stimulation and an elevation in mood. There were no effects of tea vs. coffee or caffeine dose, despite a fourfold variation in the latter. Increasing beverage strength was associated with greater increases in DBP and energetic arousal. In study 2, caffeinated beverages increased SBP, DBP, and skin conductance and lowered heart rate and skin temperature compared to water. Significant dose-response relationships to caffeine were seen only for SBP, heart rate, and skin temperature. There were significant effects of caffeine on energetic arousal but no consistent dose-response effects. Caffeinated beverages acutely stimulate the autonomic nervous system and increase alertness. Although caffeine can exert dose-dependent effects on a number of acute autonomic responses, caffeine level is not an important factor. Factors besides caffeine may contribute to these acute effects. PMID- 10837841 TI - The influence of caffeine on sustained attention: an ERP study. AB - The present study investigated the effects of caffeine on sustained attention by measuring concentration and fatigue. Event-related potentials (ERPs) and behavioral measures were recorded from 12 participants who worked continuously for approximately 10 min in a self-paced reaction task under conditions of both caffeine (250 mg) and placebo. The ERP data revealed more positive frontal P2 and parietal P3 components in the caffeine condition. However, a combination of different indices of the behavioral data did not reveal any effects of caffeine intake. These results suggest that caffeine increases arousal, thereby reducing fatigue, as was observed in the ERP results. A probable explanation for the absence of any effects of caffeine in the behavioral data can be found in the demanding properties of the task that was used, thereby supporting evidence for more pronounced effects of caffeine in suboptimal conditions. In addition, these results appeal for an increase in the use of ERPs in drug research, in order to discover possible effects on the brain which do not necessarily result in behavioral changes. PMID- 10837842 TI - Caffeine-induced increases in the brain and plasma concentrations of neuroactive steroids in the rat. AB - The effects of caffeine, a naturally occurring stimulant, on the brain and plasma concentrations of neuroactive steroids were examined in the rat. A single intraperitoneal injection of caffeine induced dose- and time-dependent increases in the concentrations of pregnenolone, progesterone, and 3alpha-hydroxy-5alpha pregnan-20-one (allopregnanolone) in the cerebral cortex. The increases were significant at a caffeine dose of 25 mg/kg and greatest (+188, +388, and +71%, respectively) at a dose of 100 mg/kg in rats killed 30 min after caffeine administration. Caffeine also increased the plasma concentrations of pregnenolone and progesterone with a dose-response relation similar to that observed in the brain, whereas the caffeine-induced increase in the plasma concentration of allopregnanolone was maximal at a dose of 50 mg/kg. Caffeine increased the plasma concentration of corticosterone, but it had no effect on the brain or plasma concentrations of 3alpha, 21-dihydroxy-5alpha-pregnan-20-one and dehydroepiandrosterone. Moreover, the brain and plasma concentrations of pregnenolone, progesterone, and allopregnanolone were not affected by caffeine in adrenalectomized-orchiectomized rats. These results suggest that neuroactive steroids may modulate the stimulant and anxiogenic effects of caffeine. PMID- 10837843 TI - Alteration of the behavioral effects of nicotine by chronic caffeine exposure. AB - The prevalence of tobacco smoking and coffee drinking place nicotine and caffeine among the most used licit drugs in many societies and their consumption is often characterised by concurrent use. The pharmacological basis for any putative interaction between these drugs remains unclear. Some epidemiological reports support anecdotal evidence, which suggests that smokers consume caffeine to enhance the effects of nicotine. This paper reviews various aspects of the pharmacology of caffeine and nicotine, in humans and experimental animals, important for the understanding of the interactions between these drugs. In particular, recent experiments are reviewed in which chronic exposure to caffeine in the drinking water of rats facilitated acquisition of self-adminstration behavior, enhanced nicotine-induced increases in dopamine levels in the shell of the nucleus accumbens and altered the dopaminergic component of a nicotine discrimination. These studies provide evidence that the rewarding and subjective properties of nicotine can be changed by chronic caffeine exposure and indicate that caffeine exposure may be an important environmental factor in shaping and maintaining tobacco smoking. PMID- 10837844 TI - The role of the dorsal hippocampal serotonergic and cholinergic systems in the modulation of anxiety. AB - A review of the literature suggests that the dorsal hippocampal serotonergic system, and, in particular, the postsynaptic 5-HT(1A) receptor, mediates an anxiogenic response, whereas endogenous dorsal hippocampal cholinergic tone mediates an anxiolytic response. Accordingly, it has been shown that direct dorsal hippocampal administration of the 5-HT(1A) receptor agonist, 8-OH-DPAT, the nicotinic receptor antagonist, mecamylamine, and the M(1) muscarinic receptor antagonist, pirenzepine, all have anxiogenic effects in rats tested in the social interaction test. It is therefore surprising that nicotine also has an anxiogenic effect in this test following dorsal hippocampal administration. However, the anxiogenic effects of mecamylamine and nicotine in the dorsal hippocampus are blocked by coadministration of the 5-HT(1A) receptor antagonist, WAY 100635, suggesting that both of these compounds act by enhancing hippocampal serotonergic transmission, thereby stimulating postsynaptic 5-HT(1A) receptors. This conclusion is supported by the observation that both nicotine and mecamylamine stimulate basal [3H]-5-HT release from dorsal hippocampal slices. A possible mechanism by which nicotinic receptor ligands modulate hippocampal 5-HT release is discussed, and it is proposed that the dorsal hippocampal serotonergic and cholinergic systems are tightly coupled and function antagonistically in the modulation of anxiety, as measured in the social interaction test. These systems are relatively unimportant in controlling behaviour on trial 1 in the plus-maze. On trial 2 in the elevated plus-maze, a model of specific phobia, the endogenous cholinergic system, nicotine, and the M(1) receptor agonist, McN-A-343, all mediate an anxiolytic effect, whereas stimulation of 5-HT(1A) receptors mediates an anxiogenic effect. It is proposed that the hippocampus may predominantly control the avoidance components of phobic anxiety, with other regions, such as the dorsomedial hypothalamus, controlling the escape components. PMID- 10837845 TI - Depressive characteristics of FSL rats: involvement of central nicotinic receptors. AB - Antidepressant effects of acute or chronic nicotine treatments in swim test immobility of Flinders sensitive line (FSL) rats, an animal model of depression, were recently demonstrated (Tizabi et al. Psychopharmacology 142:193, 1999). In the present study we sought to determine whether the antidepressant effects of nicotine could be blocked by the nicotinic antagonist, mecamylamine (MEC). Moreover, the effects of chronic nicotine treatment on [3H]cytisine binding in discrete brain regions of FSL and their control Flinders resistant line (FRL) rats were also evaluated. Adult male FSL rats were treated with MEC (0.5 mg/kg) 20 min prior to an acute or chronic nicotine administration. MEC by itself did not affect the immobility in swim test. However, it completely blocked the acute or chronic nicotine effects. Daily nicotine injection (0.4 mg/kg/day for 14 days) resulted in an increase in [3H]cytisine binding primarily in the FRL rats. An increase in nicotinic receptor binding following chronic nicotine administration is believed to reflect desensitization of these receptors. These findings, coupled with previous observation of higher basal nicotinic receptors in FSL rats, further support the involvement of central nicotinic receptors in depressive characteristics of these rats. Moreover, the data suggest therapeutic potential for selective nicotinic receptor agonists in depressive disorders. PMID- 10837846 TI - The effects of nicotine on neural pathways implicated in depression: a factor in nicotine addiction? AB - The prevalence of tobacco smoking varies considerably between different groups within the community, tobacco smoking being particularly prevalent in patients with depressive disorder. This review will focus on results, derived from animal studies, which suggest that, in addition to its primary reinforcing properties, nicotine also exerts effects in stressful environments, which may account for its enhanced addictive potential in depressed patients. It focuses on the evidence that depression sensitises patients to the adverse effects of stressful stimuli, and that this can be relieved by drugs that stimulate dopamine release in the forebrain. This mechanism, it is proposed, contributes to the increased craving to smoke in abstinent smokers exposed to such stimuli, because they become conditioned to use this property of nicotine to produce rapid alleviation of the adverse effects of the stress. The review also explores the possibility that chronic exposure to nicotine elicits changes in 5-HT formation and release in the hippocampus which are depressogenic. It is postulated that smokers are protected from the consequences of these changes, while they continue to smoke, by the antidepressant properties of nicotine. However, they contribute to the symptoms of depression experienced by many smokers when they first quit the habit. PMID- 10837847 TI - Effect of acute nicotine on Fos protein expression in rat brain during chronic nicotine and its withdrawal. AB - To study the cholinergic regulation of hypothalamic paraventricular (PVN) and supraoptic (SON) nuclei and interpeduncular nucleus (IPN) we investigated the effects of acute nicotine (0.5 mg/kg, SC, 60 min) on Fos-like immunostaining (IS) during chronic nicotine and its withdrawal in rats. Nicotine or saline was infused to rats via osmotic minipumps (4 mg/kg/day) for 7 days; on the seventh day, the minipumps were removed surgically. In control rats, acute nicotine increased Fos IS significantly in all three brain areas studied. On the seventh day of nicotine infusion this effect partially persisted in IPN but was abolished in PVN and SON. After 72-h withdrawal nicotine-induced elevation of Fos IS was similar to that of control rats in all three areas. The observed attenuation of the response to acute nicotine during constant nicotine infusion in PVN and SON may be attributable to the desensitization of nicotinic acetylcholine receptors (nAChRs) mediating the effects of nicotine in these areas or in their input areas. IPN is connected to midbrain limbic system, so in agreement with our earlier observations, it seems that limbic nicotinic receptors do not very readily desensitize during chronic nicotine infusion. These findings support the suggestions that there are differences in the level of desensitization of nAChRs. PMID- 10837848 TI - Chronic nicotine administration in the drinking water affects the striatal dopamine in mice. AB - Although tobacco contains a large variety of substances, its addictive properties are most probably due to the reinforcing actions of nicotine that motivates continued tobacco use. Animals and humans self-administer nicotine, a response that appears to involve the mesolimbic dopamine system and to be common to other abused drugs. The present article reviews animal models to administer nicotine chronically. We also describe a new animal model in which nicotine is given to mice in drinking water as their sole source of fluid. This treatment produced nicotine plasma concentrations comparable to or above those found in smokers. We found that mice withdrawn from nicotine were tolerant to the effects of nicotine challenge on striatal dopamine metabolism as well as on body temperature and locomotor activity. Furthermore, 3H-nicotine binding in the cortex and midbrain was significantly increased in mice withdrawn from nicotine. The last part of the article will focus on the effects of this chronic nicotine treatment on striatal dopamine. Dopamine and its metabolites and locomotor activity were increased in the forenoon in mice still drinking nicotine solutions. We also report recent data in which chronic nicotine administration in the drinking water enhanced the effect of dopamine receptor agonist, quinpirole, on striatal metabolism. The animal model described appears to be a relevant method for studying the mechanisms that are thought to be involved in nicotine dependence. PMID- 10837849 TI - Effects of Hypericum perforatum extract on ethanol intake, and on behavioral despair: a search for the neurochemical systems involved. AB - The present study investigated the possible involvement of sigma receptors and of serotonergic mechanisms in the effects of Hypericum perforatum extract (HPE) on immobility time in the forced swimming test (FST) and on ethanol intake in Marchigian Sardinian alcohol-preferring rats. The HPE employed was a dry extract containing 0.3% hypericin and 3.8% hyperforin. Intraperitoneal pretreatment with 20 mg/kg of the sigma receptor antagonist rimcazole (RIM), 30 min prior to HPE, completely suppressed the antiimmobility effect of HPE (3 intragastric injections of 250 mg/kg). Intracerebroventricular pretreatment with 5, 7-dihydroxytryptamine (5,7-DHT), which produced a marked depletion of brain serotonin, reduced the antiimmobility effect, although this reduction was not as pronounced as that of RIM. On the other hand, the inhibitory effect of HPE on 10% ethanol intake was modified neither by 5,7-DHT nor by RIM pretreatment. These results suggest that the antidepressant-like effect of HPE in the FST may be mediated by interaction with sigma receptors and to some extent by increased serotonergic neurotransmission. On the other hand, these mechanisms appear to be unimportant for the effect of HPE on ethanol intake. PMID- 10837850 TI - An assessment of novelty-seeking behavior in alcohol-preferring and nonpreferring rats. AB - This study examined novelty-seeking behavior in rat populations selectively bred for high and low alcohol-drinking behavior. In Experiment 1, and "odor-enhanced" novel environment produced greater behavioral activation in P compared to NP rats. In Experiment 2, the activity of high alcohol-drinking P and HAD rats was enhanced to a greater extent following the presentation of novel odors in a familiar arena, compared to the NP and LAD rats. The results suggest that, when measuring locomotor activity, alcohol-preferring rats are more reactive to novelty than their nonpreferring counterparts. Experiments 3 and 4, however, did not support the hypothesis that novelty seeking is associated with genetic vulnerability to high alcohol-drinking behavior. When measuring nose-poking behavior in response to novel odors and preference for a novel vs. a familiar chamber, behavior of the preferring lines did not differ from that of the nonpreferring lines, although P rats were more active in the place-preference paradigm. The overall results indicate that the relationship between novelty and alcohol drinking is only modestly associated, and is observed under specific conditions. Moreover, this study underscores the importance of using multiple measures when assessing complex behaviors such as novelty seeking. PMID- 10837851 TI - Reinstatement of ethanol seeking in rats: behavioral analysis. AB - The reinstatement model has been repeatedly used to study relapse to heroin- or cocaine-seeking behaviour in rats. The aim of the present study was to evaluate basic behavioral parameters of cue-induced reinstatement of ethanol seeking in a within-session paradigm. Rats were trained to respond for ethanol in an oral self administration procedure where each lever press resulted in presentation of 0.1 ml of 8% ethanol from a liquid dipper. In the reinstatement paradigm operant behaviour was first extinguished for 20 or 60 min by switching the dipper off. Then, ethanol-associated stimuli were noncontingently delivered and reinstatement of responding was assessed. Deliveries of the empty dipper, i.e., visual/auditory cues only, did not result in any reinstatement. In contrast, 15 random presentations of the dipper containing either ethanol (4-8%; v/v) or water significantly reinstated ethanol seeking. In a control self-administration experiment responding dropped to nonsignificant levels when water was substituted for ethanol. The magnitude of reinstatement did not depend on the duration of the extinction phase. These results seem to indicate that in the present paradigm reinstatement of ethanol seeking is driven by a compound stimulus including the visual/auditory cues and some nonspecific sensory properties of liquid available in the dipper. PMID- 10837852 TI - Effect of 5-HT(1B) receptor ligands on self-administration of ethanol in an operant procedure in rats. AB - Recent evidence suggests that 5-HT(1B) receptor activation modifies ethanol's reinforcing, intoxicating and discriminative stimulus effects. The present study further explored the role played by 5-HT(1A/1B) receptors by examining their influence on oral ethanol self-administration. Male Wistar rats were trained on an FR 4 schedule to obtain a reinforcer of 0.1 12% w/v ethanol solution. Once responding was stable, the effect of the 5-HT(1A/1B) agonist RU24969 alone and in combination with the 5-HT(1B) antagonist GR127935 or the 5-HT(1A) antagonists (+) WAY100135 and (+) WAY100635 was assessed. The effect of RU24969 on ethanol's pharmacokinetic profile and on operant oral saline self-administration was also examined to assess if alterations in oral ethanol self-administration were due to nonspecific effects on level pressing. For comparison, we examined the effect of another 5-HT(1A/1B) agonist, CGS12066B, on oral ethanol self-administration. Both RU24969 (0.1 to 1 mg/kg) and CGS12066B (0.1 to 1 mg/kg) significantly suppressed oral ethanol self-administration. Administration of GR127935 (1 mg/kg), significantly reversed the effects elicited by RU24969, whereas neither WAY100635 (1 mg/kg) nor (+)WAY100135 (1 mg/kg) had any effect. The effects of lower doses of RU24969 on oral ethanol self-administration were selective as oral saline self administration and blood ethanol levels were not altered by these doses. These data demonstrate that 5-HT(1B) receptor activation suppresses oral ethanol self administration. These studies provide further evidence that 5-HT(1B) receptors play a modulatory role in ethanol's behavioral effects. PMID- 10837853 TI - Lead acetate potentiates brain catalase activity and enhances ethanol-induced locomotion in mice. AB - Several reports have demonstrated that acute lead acetate administration enhances brain catalase activity in animals. Other reports have shown a role of brain catalase in ethanol-induced behaviors. In the present study we investigated the effect of acute lead acetate on brain catalase activity and on ethanol-induced locomotion, as well as whether mice treated with different doses of lead acetate, and therefore, with enhanced brain catalase activity, exhibit an increased ethanol-induced locomotor activity. Lead acetate or saline was injected IP in Swiss mice at doses of 50, 100, 150, or 200 mg/kg. At 7 days following this treatment, ethanol (0.0, 1.5, 2.0, 2.5, or 3.0 g/kg) was injected IP, and the animals were placed in the open-field chambers. Results indicated that the locomotor activity induced by ethanol was significantly increased in the groups treated with lead acetate. Maximum ethanol-induced locomotor activity increase was found in animals treated with 100 mg/kg of lead acetate and 2.5 g/kg of ethanol. Total brain catalase activity in lead-pretreated animals also showed a significant induction, which was maximum at 100 mg/kg of lead acetate treatment. No differences in blood ethanol levels were observed among treatment groups. The fact that brain catalase and ethanol-induced locomotor activity followed a similar pattern could suggest a relationship between both lead acetate effects and also a role for brain catalase in ethanol-induced behaviors. PMID- 10837854 TI - Withdrawal symptoms in a long-term model of voluntary alcohol drinking in Wistar rats. AB - Long-term voluntary alcohol drinking with repeated alcohol deprivation episodes has been suggested as animal model for some aspects of alcoholism. Using a radiotelemetric system, the present study investigated the occurrence of withdrawal symptoms in long-term voluntarily alcohol drinking Wistar rats with (repeated alcohol deprivation group) and without (first alcohol deprivation group) prior alcohol deprivation experience. Six days after transmitter implantation, alcohol bottles were removed, and returned 4 days later. Alcohol deprivation induced hyperlocomotion in both groups. In the repeated alcohol deprivation group, hyperlocomotion was increased at the beginning of the alcohol deprivation phase and decreased during the following dark phase, suggesting that removal of the alcohol bottles might have become a conditioned withdrawal stimulus for this group. Both groups showed an enhanced alcohol intake after representation of alcohol bottles compared to preabstinence intakes (alcohol deprivation effect). However, alcohol intake of the repeated alcohol deprivation group was significantly increased compared to the first alcohol deprivation group at the end of the experiment. It is concluded that repeated alcohol deprivation experience might promote the development of alcohol addiction because of its latent stimulating effect on alcohol drinking that can be unveiled by (presumably mildly stressful) experimental situations. PMID- 10837855 TI - Prolonged changes in neurochemistry of dopamine neurones after chronic ethanol consumption. AB - The effects of 3 weeks of chronic ethanol consumption in mice on brain concentrations and turnover of monamine transmitters was examined. The measurements were made at 24 h, 6 days and 2 months after cessation of the ethanol intake to examine changes that might be relevant to relapse drinking. Increases in noradrenaline and dopamine concentrations, and decreases in the ratios of dopamine metabolites to dopamine, were seen in ventral tegmental tissue at 24 h after alcohol consumption. Increased noradrenaline was also evident at the 6-day interval, but no other changes were seen at this time. At the 2-month interval, the ventral tegmentum from ethanol-treated animals showed decreases in metabolite/dopamine ratios. No changes were seen in 5-hydroxytryptamine or its metabolite. In striatal tissue, none of these changes were seen, but at 24 h decreases occurred in the content of dopamine and its metabolites and a decrease in 5-hydroxyindoleacetic acid. The results indicate changes occur in monoamine turnover in the VTA as long as 2 months after cessation of chronic ethanol consumption; such changes may be related to the prolonged nature of alcohol dependence. PMID- 10837856 TI - Blood cholesterol levels of 32-year-old alcohol consumers are better than of nonconsumers. AB - Blood cholesterol levels are expected to be important factors in the causal pathway between alcohol consumption and CHD. The relation between alcohol consumption and blood cholesterol levels is investigated in 130 men and 145 women aged 32.4 years old (+/-1.0), from the Amsterdam Growth and Health Longitudinal Study. When controlled for gender, cholesterol levels at age 13.1 years, and lifestyle at adult age (smoking, physical activity, dietary habits), no significant differences were found for total cholesterol (TC) levels between alcohol consumers and nonconsumers. Serum high-density lipoprotein (HDL) cholesterol levels were 0.12 mmol/l higher in subjects consuming >/=100 grams of alcohol per week than in nonconsumers (p < 0.05). Regression coefficients of subjects consuming 10 to 50, or 50 to 100 g alcohol per week did not differ statistically from those of nonconsumers. The positive relation between alcohol consumption and serum HDL was modified by smoking (found in nonsmokers, but not in smokers). No differences between beer, wine, and spirits were found for their relation with serum HDL. In conclusion, 32.4-year-old nonsmoking subjects who consumed >/=100 g of alcohol per week had improved HDL levels compared with nonconsumers, whereas the protective effect of drinking smaller amounts of alcohol did not reach statistical significance. PMID- 10837857 TI - Motivations to control drinking behavior in abstainers, moderate, and heavy drinkers. AB - Motives and motivational concepts to control quantity (Q) and frequency (F) of alcohol consumption were investigated in 192 participants using a questionnaire of motives to control drinking (MCD-Q/F) developed earlier. The daily quantity of consumption was assessed by the "30 day by beverage questionnaire." Participants were grouped into heavy or moderate drinkers (limits: females 40, males 60 gram alcohol/day) and abstainers. Analyses of motives showed the importance of car driving, controlling appetite/desire, and prevention of impaired mental performance. The relative impact of motivational concepts were analyzed by cluster analysis of motives revealing three clusters: 1) emotional concepts/fear of addiction, 2) physiological sensations/interaction with concrete actual plans, 3) cognitive concepts (plans, self-control, no impairment/no desire). MANOVA of MCD-Q/F indicate highest scores for moderate drinkers and significant lower scores in heavy drinkers in controlling frequency of drinking (MCD-F). Group effect for emotional concepts failed significance, higher scores of physiological concepts were found in imbibers compared to abstainers, and scores of cognitive concepts were lower in heavy drinkers. Results indicate that for imbibers a reduced motivation to control frequency of drinking as well as low scores in cognitive concepts and a trend to high scores in emotional concepts are associated with heavy drinking. PMID- 10837858 TI - Different effects of nabilone and cannabidiol on binocular depth inversion in Man. AB - The physiological and pathophysiological roles of the central nervous endogenous cannabinoid system are not completely understood, but still represent a challenge in basic neurobiological, cognitive, and psychiatric research. The system has been implicated in the pathogenesis of schizophrenia. Binocular depth inversion, an illusion of visual perception, provides a model of impaired perception during psychotic states. Using this model the effects of nabilone, a psychoactive synthetic 9-trans-ketocannabinoid, and of cannabidiol, the main natural component of herbal cannabis, and a combined application of both substances on binocular depth inversion and behavioural states were investigated in nine healthy male volunteers. The time course of the effects of both substances on binocular depth inversion was analysed after oral administration using three different groups of natural stimuli. A significant impairment of binocular depth perception was found when nabilone was administered, but combined application with cannabidiol revealed somewhat reduced effects on binocular depth inversion. The influence of psychoactive cannabinoids on this perceptual model and the role of the endogenous cannabinoid system in visual information processing are discussed. PMID- 10837859 TI - Permissive role of dopamine D(2) receptors in the hypothermia induced by delta(9) tetrahydrocannabinol in rats. AB - Cannabinoids produce analgesia, hypomotility, catalepsy, cognitive deficits and positive reinforcement. Moreover, Delta(9)-tetrahydrocannabinol (9-THC) and synthetic cannabinoids stimulate dopaminergic neurons and increase dopamine release in different brain areas. In order to clarify the role of endogenously released dopamine in the hypothermic response to cannabinoids, the effect of D(1) and D(2) dopamine receptor agonists and antagonists on Delta(9)-THC-induced hypothermia was studied in rats. Delta(9)-THC (2.5 and 5 mg/kg intraperitoneally [IP]) decreased body temperature in a dose-related manner. This effect was antagonized not only as expected by the CB(1) cannabinoid receptor antagonist SR 141716A (0.5 mg/kg, IP) but also, unexpectedly, by the dopaminergic D(2) receptor antagonists S(-)-sulpiride (5 and 10 mg/kg, IP) and S(-)-raclopride (1 and 3 mg/kg, IP). Conversely, the hypothermic effect of Delta(9)-tetrahydrocannabinol was potentiated by the D(2) dopamine receptor agonists (-)-quinpirole (0.025 and 0.500 mg/kg, SC) and (+)-bromocriptine (0.5 and 1 mg/kg, IP). In contrast, the Delta(9)-THC-induced hypothermic effect was not modified by either by the D(1) dopamine agonist SKF 38393 (10 mg/kg SC) or by the D(1) dopamine antagonist SCH 23390 (0.5 mg/kg SC). These results suggest that the D(2) dopamine receptors have a permissive role in the hypothermic action of cannabinoids. PMID- 10837860 TI - Lack of specific effects of selective D(1) and D(2) dopamine antagonists vs. risperidone on morphine-induced hyperactivity. AB - In the present study, three different dopamine antagonists were challenged in order to counteract hyperactivity induced by 50 mg/kg of morphine. A wide range of doses of morphine (50, 25, 12.5, 6.25, or 3.12 mg/kg) were evaluated on spontaneous locomotor activity. A significant increase was observed only with the two higher doses tested (25 and 50 mg/kg). No decrease was found with any of the doses used at any period of time. After analyzing doses of SCH 23390 (0.5, 0.1, and 0.05 mg/kg), raclopride (0.5, 0.25, and 0.125 mg/kg) and risperidone (0.1, 0.05, and 0.025 mg/kg) administered alone, only the 0.5 mg/kg dose of SCH 23390 decreased locomotor activity. The three compounds counteracted morphine-induced hyperactivity, but with SCH 23390 it was only achieved with the dose of 0.5 mg/kg, which also decreased spontaneous locomotor activity and induced catalepsy. On the other hand, raclopride and risperidone neutralized morphine-induced hyperactivity at doses that did not affect locomotor activity, although the former induced catalepsy when administered with morphine. It is concluded that although the blockade of D(1) and D(2) DA receptors decreases morphine-induced hyperactivity, this action is not specific, contrary to the action of risperidone, which counteracts this hyperactivity without any other motor effects. PMID- 10837861 TI - Stereoselective morphine-like discriminative properties of a new alkylaminonaphthalenic derivative. AB - The morphine-like properties of a series of aminoalkyl- and cycloalkylamino naphtalenic derivatives of 17-methyl-17-azaequilenine were studied in rats trained to discriminate morphine (5.6 mg/kg IP) from vehicle in a two-lever operant behavioral procedure reinforced by water access. It was found that one of the compounds tested (i.e., A8; 1-ethyl-1-hydroxy-1-[2-(6-hydroxynaphthyl)]-2 methyl-3- dimethylaminopropane) fully generalized for the morphine stimulus. The discriminative effects of A8 were stereospecific, as indicated by the fact that (+)-(1R,2R)-A8 was three times more potent than the racemic compound and that the (-)-(1S,2S) enantiomer was completely inactive. (+)-(1R,2R)-A8 generalization for the morphine cue was inhibited by naloxone. None of the other five derivatives examined generalized for the morphine stimulus. In conclusion, the naphthalenic structure is a source of compounds with stereospecific and naloxone-reversible morphine-like properties. PMID- 10837862 TI - Lack of crosstolerance between morphine and morphine-6-glucuronide as revealed by locomotor activity. AB - Morphine-6beta-glucuronide is a major metabolite of morphine. We wanted to examine whether the effects related to opiate CNS stimulation could be mediated by different receptors for morphine and M6G by studying the development of crosstolerance between these two drugs. The effect studied was locomotor activity in C57BL/6JBom mice. We observed a dose-dependent development of tolerance to daily injections of morphine, with 20 micromol/kg giving the most rapid development of tolerance, apparent already on the second day of treatment. This was also observed for the same dose of M6G. Crosstolerance to M6G was measured both after 1 day pretreatment and 7 days pretreatment with morphine 20 micromol/kg, while the crosstolerance to morphine was tested only after 1 day pretreatment with M6G (20 micromol/kg). Lack of crosstolerance towards M6G after 1 day of morphine pretreatment was observed, whereas crosstolerance to M6G was observed after 7 days of exposure to morphine pretreatment. Crosstolerance after M6G pretreatment to morphine was observed. It was concluded that the main part of the effect caused by M6G was mediated through a specific M6G receptor. PMID- 10837863 TI - Effects of serotonergic manipulations on the behavioral sensitization and disinhibition associated with repeated amphetamine treatment. AB - This study investigated the effects of repeated amphetamine treatment on locomotor activity and behavioral inhibition in the elevated plus-maze, and the influence of serotonin (5-HT) neurotransmission on these behaviors. Acute administration of amphetamine (1.0 mg/kg subcutaneously [SC]) stimulated locomotor activity, which was attenuated by acute citalopram (5.0 mg/kg SC) pretreatment. Repeated daily treatment with amphetamine (15 days) sensitized the rats to the amphetamine-induced locomotor stimulation. Acute pretreatment with the 5-HT precursor l-5-hydroxytryptophan (5-HTP; 25 mg/kg IP) or chronic treatment with the selective 5-HT reuptake inhibitor citalopram (5.0 mg/kg SC, twice daily), did not alter the expression of amphetamine-induced locomotor sensitization. In the elevated plus-maze, animals subjected to repeated amphetamine treatment expressed behavioral disinhibition after amphetamine exposure (1.0 mg/kg SC; -35 min), which was antagonized both by acute 5-HTP and chronic citalopram treatment. In summary, these findings suggest that behavioral sensitization to amphetamine is associated with amphetamine-induced behavioral disinhibition, and that acute 5-HTP as well as chronic citalopram treatment counteract the expression of amphetamine-induced behavioral disinhibition, but not locomotor sensitization. It appears likely that the antagonistic effects of 5 HTP and citalopram on behavioral disinhibition derive from a drug-induced facilitation of brain 5-HT neurotransmission. PMID- 10837864 TI - Absence of cocaine-induced place conditioning in serotonin 1B receptor knock-out mice. AB - A large body of evidence suggests that genetic factors may affect the reinforcing properties of drugs of abuse. This study investigated the involvement of the serotonin 1B (5-HT1B) receptor in modulating cocaine-induced place conditioning by comparing the response of 5-HT1B receptor gene knock-out mice with wild type 129/Sv-ter mice. The rewarding effects of various doses of cocaine (0, 2.5, 5, 10, 20, and 40 mg/kg) were examined in both strains. Results clearly show that 5 HT1B receptor knock-out mice failed to display a conditioned place preference for stimuli paired with cocaine while wild type mice exhibited a conditioned place preference for the compartment paired with cocaine (5 and 20 mg/kg). As other studies showed that 5-HT1B knock-out mice self-administer cocaine, these results suggest a dissociation between the psychologic state linked to self administration and the one measured in conditioned place preference. PMID- 10837865 TI - The retrograde tracer fluoro-gold interferes with the expression of fos-related antigens. AB - The retrograde tracer, fluoro-gold (FG) has been used in combination with immediate early gene (IEG) immunohistochemistry to identify neural circuits activated by pharmacological, physiological or behavioral manipulations. However, since FG has been shown to be toxic to cell bodies, axons and terminals at the injection site, the question arises as to whether FG alters the detection of IEG products. To examine this question, FG was microiontophoresed unilaterally into the nucleus accumbens (NAc) of rats and Fos-related antigens (FRAs) were examined in both hemispheres 12 days later. Approximately half as many FRA-positive nuclei were observed in the tracer-injected NAc as were found in the contralateral NAc. Similar results were observed in the ventral subiculum of the hippocampus and the basolateral and central amygdaloid nuclei, but not in the lateral septum or lateral habenula. These results suggest that FG microiontophoresed into the NAc interferes with the expression of FRAs at the injection site and also at other ipsilateral limbic sites. PMID- 10837866 TI - The use of TaqMan RT-PCR assays for semiquantitative analysis of gene expression in CNS tissues and disease models. AB - TaqMan reverse transcription polymerase chain reaction (RT-PCR) is a recently developed technique which allows the measurement of an accumulating PCR product in real time. In the present study we have validated the use of TaqMan RT-PCR for mRNA localisation studies in human and rat tissues, and for the investigation of gene expression changes in CNS animal models. In human brain, D(2) receptor mRNA was enriched in caudate nucleus and putamen, whilst in rat brain, highest levels of D(2) receptor mRNA expression were observed in striatum and nucleus accumbens, consistent with the known distribution of this receptor in basal ganglia. In a rat model of permanent middle cerebral artery occlusion (pMCAO), endogenous interleukin-1 receptor antagonist (IL-1ra) mRNA was upregulated over 30-fold at 24 h post-lesion in both striatum and cortex ipsilateral to artery occlusion. Brain-derived neurotrophic factor (BDNF) mRNA was transiently upregulated 3.7 fold at 3 h, but not at 24 h or 3 days after induction of cortical spreading depression (CSD) in rats. Our observations in these two animal models using TaqMan RT-PCR were consistent with previous reports using other techniques. In conclusion, TaqMan RT-PCR assays provide a rapid and reliable method for semi quantitative analysis of gene expression in the nervous system. PMID- 10837867 TI - Optical light scatter imaging of cellular and sub-cellular morphology changes in stressed rat hippocampal slices. AB - Optical imaging, such as transmission imaging, is used to study brain tissue injury. Transmission imaging detects cellular swelling via an increase in light transmitted by tissue slices due to a decrease in scattering particle concentration. Transmission imaging cannot distinguish sub-cellular particle size changes from cellular swelling or shrinkage. We present an optical imaging method, based on Mie scatter theory, to detect changes in sub-cellular particle size and concentration. The system uses a modified inverted microscope and a 16 bit cooled CCD camera to image tissue light scatter at two angles. Dual-angle scatter ratio imaging successfully discriminated latex microsphere suspensions of differing sizes (0.6, 0.8, 1 and 2 microm) and concentrations. We applied scatter imaging to hippocampal slices treated with 100 microM N-methyl-D-aspartate (NMDA) to model excitotoxic injury or -40 mOsm hypotonic perfusion solution to cause edema injury. We detected light scatter decreases similar to transmission imaging in the CA1 region of the hippocampus for both treatments. Using our system, we could distinguish between NMDA and hypotonic treatments on the basis of statistically significant (P<0.0003) differences in the scatter ratio measured in CA1. Scatter imaging should be useful in studying tissue injuries or activity resulting in brain tissue swelling as well as morphological changes in sub cellular organelles such as mitochondrial swelling. PMID- 10837868 TI - 'Specific' oligonucleotides often recognize more than one gene: the limits of in situ hybridization applied to GABA receptors. AB - As exquisite probes for gene sequences, oligonucleotides are one of the most powerful tools of recombinant molecular biology. In studying the GABA receptor subunits in the neonatal hippocampus we have used oligonucleotide probes in in situ hybridization and cloning techniques. The oligonucleotides used and assumed to be specific for the target gene, actually recognized more than one gene, leading to surprising and contradictory results. In particular, we found that a GABA(A)-rho specific oligonucleotide recognized an abundant, previously unknown, transcription factor in both in situ and library screening, while oligos 'specific' for GABA(A) subunits were able to recognize 30 additional unrelated genes in library screening. This suggests that positive results obtained with oligonucleotides should be interpreted with caution unless confirmed by identical results with oligonucleotides from different parts of the same gene, or cDNA library screening excludes the presence of other hybridizing species. PMID- 10837869 TI - Evaluation of an optimal temperature for brain storage in delayed 2, 3,5 triphenyltetrazolium chloride staining. AB - This paper presents the determination of an optimal temperature for delayed 2,3,5 triphenyltetrazolium chloride (TTC) staining. Twenty-one rats were subjected to right middle cerebral artery embolic stroke and sacrificed 96 h following ischemia. The brains were harvested and stained immediately after sacrifice or stored for 8 h at 21-23 degrees C or 4 degrees C, respectively. The stained sections were scanned and infarct volume calculated. The quality of staining, distinction of borders between infarcted and non-ischemic tissue and ease of differentiating ischemic tissue in colored and grayscale images was assessed. The present study indicates that results of TTC obtained immediately after animal sacrifice, or delayed TTC staining while storing the brains at room temperature or 4 degrees C are comparable. PMID- 10837870 TI - Linear inverse filtering improves spatial separation of nonlinear brain dynamics: a simulation study. AB - We examined topographic variations in nonlinear measures based on scalp voltages, which were generated by two simulated current dipoles each placed in a different hemisphere of a spherical volume conductor (three-shell model). Dipole dynamics were that of a three-torus and the x-component of the Lorenz-system and scalp voltage were calculated for a configuration of 29 electrode positions. Although estimates for correlation dimension D2 and Lyapunov exponent L1 were close to the theoretical values for the original time series, the simulated scalp voltage data showed almost no topographic resolution of dipole positions. In order to enhance topographic differentiation, we constructed linear inverse filters, to focus on brain activity from a specified brain region. It turned out that the nonlinear measures for the inversely filtered time series were much closer to the expected values (with respect to the location of the dipoles used in the simulation) than when using unfiltered data. Our preliminary results indicate that inverse filtering can improve the topographic resolution of nonlinear scalp EEG estimates. PMID- 10837871 TI - A digital averaging method for removal of stimulus artifacts in neurophysiologic experiments. AB - Stimulation artifacts can greatly complicate the evaluation of short-latency responses in experiments in which anterograde stimulation techniques are used to investigate connections between different brain regions. For an experiment involving recording of the responses in the primate substantia nigra pars reticulata to stimulation at various sites in the striatum, a digital averaging technique was developed for removing stimulation artifacts from traces of neuronal signals. In the first of two stages of this procedure, an estimate of the average stimulus artifact is calculated from traces of multiple stimulations at the same site. In the second step, the resulting average stimulation artifact (after time- and amplitude-optimization) is subtracted from individual data segments that contain the artifact. The data presented here demonstrate that this technique, applied off-line, is highly effective in removing artifacts, and uncovering neuronal potentials superimposed on the artifact. Faster computers and optimized software may make on-line application of this technique feasible. PMID- 10837872 TI - Development and characterization of human and mouse specific antibodies to CuZn superoxide dismutase (SOD1). AB - Mutations in the copper/zinc superoxide dismutase (SOD1) gene are associated with 15-20% of the familial forms of motor neuron disease. Mice where a transgene has been incorporated that encodes for the human SOD1 mutation develop a form of motor neurone disease that closely resembles human forms of this disease. We have produced and characterized species-specific antibodies to epitopes in the SOD1 protein, amino acids 25-37, a region that distinguishes between the human and the mouse species of SOD1. The antisera generated were unable to immunoprecipitate the mouse or the human forms of SOD1 from tissue extracts unless the homodimeric complex of SOD1 was denatured. As SOD1 exists as a homodimeric complex in the cytoplasm of cells, this suggests that amino acids in position, 25-37 are close to the dimeric interface of SOD1. PMID- 10837873 TI - Quantitating silver-stained neurodegeneration: the neurotoxicity of trimethlytin (TMT) in aged rats. AB - This report describes the development of a histoanalytical procedure to measure the degree of neurodegeneration produced by the organometal toxicant trimethyltin (TMT). Based on a previous, non-quantitated experiment we hypothesized that the same dose of TMT would produce greater damage in animals of increasing age. Male rats aged 6, 12, 18, or 24 months at the time of dosing were given either 4.5 mg/kg TMT or saline (i.p.). One month after dosing, rats were perfused and their brains removed and processed to selectively silver-impregnate degenerating cell bodies as well as axon terminals and dendrites. Neurodegeneration was most prominent in the hippocampi (especially CA1 stratum radiatum) of TMT-treated rats, but not in the controls. Computer-assisted counting of the silver grains marking damage indicated greater neurotoxicity from the same dose of TMT when given to the older animals. Thus the grain density in the 6-month-old TMT-treated rats was not significantly elevated from the 6-month-old controls (P>0.10). The 12-month-old TMT-treated rats had significantly increased grain densities compared to their controls (P<0.05), but still larger increases of grain counts were observed in the 18- and 24-month-old rats (both P-values<0.01). Our findings with TMT are similar to previous, but nonquantitative, reports that the neurotoxic effects of kainic acid and methionine sulfoximine were also greater in older rats. An increased sensitivity to neurotoxicants might help explain the apparently spontaneous degeneration of cortical neurons in aging and in the neurological diseases of old age. The method we report here for quantitation of silver grains marking neurodegeneration should be adaptable to a wide range of histologically-based neurotoxicology investigations. PMID- 10837874 TI - Multiple site silicon-based probes for chronic recordings in freely moving rats: implantation, recording and histological verification. AB - This paper describes the procedure of assembling a miniature microdrive and silicon probe system for surgical implantation into the adult rat brain. Successful recordings of single and multiunit activity with parallel depth profiles of spontaneous and evoked field potentials are shown. The procedure for histological verification of the position of the silicon probe is described. PMID- 10837875 TI - The detection and quantitation of inflammation in the central nervous system during experimental allergic encephalomyelitis using the radiopharmaceutical 99mTc-RP128. AB - RP128 is a novel agent which readily chelates 99mTc to form a radiopharmaceutical which binds in vivo to the tuftsin receptor located specifically on neutrophils and monocyte-macrophages, therefore removing the need for in vitro cell labelling prior to intravenous administration. We have assessed the ability of 99mTc-RP128 to detect central nervous system (CNS) inflammation in experimental allergic encephalomyelitis (EAE), an animal model of the human disease multiple sclerosis. The radiopharmaceutical was recorded at significantly increased levels in all EAE diseased CNS tissues, compared to normal and control samples, at 0.5, 1 and 3 h post-injection using a dual radioisotope technique to correct for non extravasated tracer (P<0.05). Moreover, extravascular accumulation of the agent could be clearly demonstrated in inflammatory tissues with minimal loss of sensitivity when the secondary isotopic correction for blood volume was omitted. In addition, 99mTc-RP128 successfully monitored glucocorticoid suppression of inflammation (P<0.05), recording a typical dose-response to increasing steroid concentration. Clearly, 99mTc-RP128 can quantitatively detect CNS inflammation and assess responses to therapy indicating potential value as an imaging agent both clinically and as a research aid. Furthermore, the rapid in vivo labelling by 99mTc-RP128 of specific inflammatory cells combined with the ability to monitor the progress of anti-inflammatory therapeutics may recommend the agent for use in a variety of inflammatory conditions. PMID- 10837877 TI - Seasonal mood patterns in a northeastern college sample. AB - Given that seasonality can be conceptualized along a developmental continuum, it is surprising that the college population has been largely excluded from previous epidemiological research on Seasonal Affective Disorder (SAD). In this study, college students completed the Seasonal Pattern Assessment Questionnaire (SPAQ) and Beck Depression Inventory twice during either the Fall or Spring academic semester. SAD prevalence rates were similar to those of nearby adult samples. The SPAQ demonstrated high test-retest reliability when administered during different seasons; however, gender differences only emerged during winter SPAQ administrations. PMID- 10837878 TI - Anxiety, depression and psychosis in vascular dementia: prevalence and associations. AB - BACKGROUND: Little is known about psychiatric symptoms in Vascular dementia (VaD). METHOD: 92 patients with VaD, and 92 patients with Alzheimer's disease (AD) are reported. The evaluation included standardised measures of mood and psychosis. RESULTS: 72% of VaD patients and 38% of those with AD had two or more anxiety symptoms. VaD patients with severe dementia (94%) were the most likely to be anxious. Depression was also significantly more common in VaD patients (19% vs. 8%) whereas psychotic symptoms were prevalent in both dementias. CONCLUSION: Psychiatric symptoms are common in VaD, especially in patients with moderate or severe dementia. Rigorous assessment of psychiatric symptoms in VaD should be part of good clinical practice. PMID- 10837879 TI - Suicidal behavior in bipolar mood disorder: clinical characteristics of attempters and nonattempters. AB - OBJECTIVE: Bipolar Disorder is associated with a higher frequency of attempted suicide than most other psychiatric disorders. The reasons are unknown. This study compared bipolar subjects with a history of a suicide attempt to those without such a history, assessing suicidal behavior qualitatively and quantitatively, and examining possible demographic, psychopathologic and familial risk factors. METHODS: Patients (ages 18 to 75) with a DSM III-R Bipolar Disorder (n = 44) diagnosis determined by a structured interview for Axis I disorders were enrolled. Acute psychopathology, hopelessness, protective factors, and traits of aggression and impulsivity were measured. The number, method and degree of medical damage was assessed for suicide attempts, life-time. RESULTS: Bipolar suicide attempters had more life-time episodes of major depression, and twice as many were in a current depressive or mixed episode, compared to bipolar nonattempters. Attempters reported more suicidal ideation immediately prior to admission, and fewer reasons for living even when the most recent suicide attempt preceded the index hospitalization by more than six months. Attempters had more lifetime aggression and were more likely to be male. However, attempters did not differ from nonattempters on lifetime impulsivity. LIMITATIONS: The generalizability of the results is limited because this is a study of inpatients with a history of suicide attempts. Patients with Bipolar I and NOS Disorders were pooled and a larger sample is needed to look at differences. We could not assess psychopathology immediately prior to the suicide attempt because, only half of the suicide attempters had made attempts in the six months prior to admission. Patients with current comorbid substance abuse were excluded. No suicide completers were studied. CONCLUSIONS: Bipolar subjects with a history of suicide attempt experience more episodes of depression, and react to them by having severe suicidal ideation. Their diathesis for acting on feelings of anger or suicidal ideation is suggested by a higher level of lifetime aggression and a pattern of repeated suicide attempts. PMID- 10837880 TI - Fluoxetine versus sertraline and paroxetine in major depression: tolerability and efficacy in anxious depression. AB - BACKGROUND: Major depression with high levels of anxiety (anxious depression) is a common subtype of depression associated with greater psychosocial impairment and poorer response to antidepressant treatment. It is unclear whether in this population there are differences in efficacy or tolerability across selective serotonin reuptake inhibitors. For this reason, using head-to-head acute treatment comparison, we compared efficacy and tolerability of fluoxetine, sertraline, and paroxetine among depressed patients with high levels of anxiety. METHODS: Patients (N = 108) with DSM-IV major depression and high levels of anxiety (a HAM-D-Anxiety/Somatization Factor score > or =7) were randomized to fluoxetine, sertraline, or paroxetine treatment in a double-blind fashion. Changes in overall depression and anxiety were assessed. RESULTS: Patients demonstrated similar baseline-to-endpoint improvement in HAM-D-17 and HAM-D Anxiety/Somatization Factor scores. Patients also demonstrated similar change over-time improvement in HAM-D-17 and HAM-D-Anxiety/Somatization Factor scores, except at week one where fluoxetine- and sertraline-treated patients had statistically significantly greater improvement than paroxetine-treated patients in the HAM-D-Anxiety/Somatization Factor score. There were no significant differences across treatments in percentages of patients with substantial emergence, any worsening, or improvement at endpoint in individual HAM-D Items 9 (agitation), 10 (psychic anxiety), and 11 (somatic anxiety). Overall, all treatments were well tolerated. CONCLUSION: These data showed no significant differences in efficacy and tolerability of fluoxetine, sertraline, and paroxetine in patients with high levels of baseline anxiety symptoms during the acute treatment of major depression. Each treatment was similarly effective in improving depression in this subtype of patients with anxious depression. PMID- 10837881 TI - Risk factors for depression in later life; results of a prospective community based study (AMSTEL). AB - BACKGROUND: Depression in the elderly was found to be associated with a variety of risk-factors in cross sectional designs. Based on the vulnerability-stress model, etiologic pathways for depression have been suggested, with vulnerability modifying the effect of stress factors. The current prospective study tests an etiologic model for depression incidence, by assessing modifying effects of three types of vulnerability: genetic/familial vulnerability, organic vulnerability, and environmental vulnerability. METHODS: 1940 non-depressed community-living elderly were interviewed at baseline, and at follow-up three years later. Bivariate and multivariate relationships between risk factors and incident depression (GMS-AGECAT) were studied. RESULTS: Higher age, personal history of depression, death of spouse, health related factors and comorbid organic or anxiety syndrome showed significant bivariate associations with depression incidence. In multivariate analysis, the effect of stress factors on incident depression was not modified by a genetic/familial vulnerability, nor by an organic vulnerability. Effect modification by environmental factors was however evident; having a marital partner, and if unmarried having social support, significantly reduced the impact of functional disabilities on the incidence of depression. LIMITATIONS: The study consisted of two measurements with a three years interval, depressive episodes with a short duration may be under represented. CONCLUSIONS: In the elderly, the effect of stress on incident depression is modified by environmental vulnerability. No evidence was found of effect modification by either genetic/familial or organic vulnerability. The results have implications for both recognition and treatment of late-life depression. PMID- 10837882 TI - Age of onset in affective disorder: its correlation with hereditary and psychosocial factors. AB - BACKGROUND: Affective disorders probably have a multifactorial aetiology, both biological and psychosocial factors may be of importance at onset as well as at relapses. The aim of the study was to investigate how the age of onset of bipolar and unipolar disorder relates to family history of affective disorder, early parental separation and life events. A second purpose of this study was to analyze the importance of life events preceding the first and subsequent episodes of affective disorder. METHODS: The case records of 282 patients (161 females/121 males; mean age 56) were investigated. They all had a DSM-IV based diagnosis of either bipolar I/II (67%) or unipolar (33%) disorder. Variables, such as family history, early parental loss and life events according to Paykel life events scale, were examined. RESULTS: We found a significantly lower age of onset in bipolar patients with a family history of affective disorder (28.9 vs. 33.9 years). Bipolar patients with preceding life events had a higher age of onset (33.1 vs. 28.3 years). Moreover, bipolar patients with heredity, had less life events at onset. For the bipolar, as well as the unipolar group, life stressors more frequently preceded the first episode of affective disorder than the subsequent episodes. LIMITATIONS: The major limitation of this study is the retrospective approach, with e.g. difficulties to decide whether a life event plays a role in aetiology of affective disorder or is its consequence. CONCLUSIONS: Bipolar patients with high constitutional vulnerability have an earlier age of onset and need less stress factors to become ill. Better knowledge about the stress- and the vulnerability-factors in affective disorder might contribute to development of individually tailored therapeutic strategies in future. PMID- 10837883 TI - The treatment of depression in UK general practice: selective serotonin reuptake inhibitors and tricyclic antidepressants compared. AB - BACKGROUND: Antidepressants are commonly prescribed by general practitioners as treatment for depression. Controversy exists as to the effectiveness in everyday use of the older tricyclic antidepressants (TCAs) when compared to the newer selective serotonin reuptake inhibitors (SSRIs). AIM: To investigate the patterns of current prescribing of antidepressants for the treatment of depression and compare TCAs with the newer SSRIs. METHOD: The study population was patients attending 151 computerised general practices from throughout the United Kingdom between 1991 and 1996. Patients with new prescriptions for antidepressants and a diagnosis of depression were identified. Age and gender distributions, prescribed doses and drop-out rates were investigated. RESULTS: During the study period 9.8% of patients received a prescription for an antidepressant, there was a 40% increase in the prescribing rate of TCAs and a 460% increase in SSRI prescribing. TCAs were initially prescribed in sub-therapeutic doses. More than 50% of patients ceased taking their antidepressants within 6 weeks of starting treatment. Fluoxetine and paroxetine were more likely to be prescribed for a therapeutic period than were other antidepressants. CONCLUSIONS: General practitioners should prescribe a therapeutic dose of antidepressant for a recognised therapeutic period to ensure that patients with depression receive the most effective treatment. PMID- 10837884 TI - Alpha-1-acid glycoprotein in major depressive disorder. Relationships to severity, response to treatment and imipramine plasma levels. AB - BACKGROUND: Increased plasma levels of alpha-1-acid glycoprotein (AGP) were reported in major depressive disorder. However, the relationship between AGP levels, severity of depression, treatment response and antidepressant levels are still unclear. METHODS: Plasma AGP levels were measured in 36 subjects with major depressive disorder before and after a 6-week treatment with imipramine and in 30 controls. Free imipramine plasma levels of depressed patients were measured at 6 weeks. Comparative analysis between depressed patients and controls, between non responders (N = 12) and responders (N = 24), and between severely depressed patients (N = 14) and moderately depressed patients (N = 22) were made. RESULTS: Depressed patients had significantly higher mean values of AGP than control subjects. Imipramine non-responders and specially severely depressed patients had significantly greater increases of AGP levels during treatment than other depressed subgroups. There was no correlation between baseline AGP levels and severity of depression or free imipramine levels. LIMITATIONS: The most significant limitations of this study are the small sample size and the fact that all the subjects were out-patients. Results should not be generalized to in patient populations. CONCLUSIONS: Depressed patients showed high baseline concentrations of AGP. AGP levels did not predict either free imipramine plasma levels or differential response after 6 weeks of treatment with imipramine. A greater increase of AGP during treatment was associated with severity of depression and treatment non-response. CLINICAL IMPLICATIONS: The relationship between high plasma levels of AGP, severity of depression and lack of treatment response is clarified. The influence of imipramine levels is minimized. PMID- 10837885 TI - A case-control study of tattoos in young suicide victims as a possible marker of risk. AB - BACKGROUND: The goal was to examine tattooing in suicides, as tattoos have been associated with several risk factors for suicide. METHOD: A chart review of a three-year sample of 134 consecutive suicides in Mobile County, Alabama, was conducted. The prevalence of tattoos was compared between young (<30) white suicides and accidental deaths matched for age, gender and race, in a case control study. RESULTS: Tattoos were found in 21% of suicides. Fifty-seven percent of young white suicides were tattooed compared to 29% of matched accidental deaths. LIMITATIONS: Findings are preliminary due to the small sample size. The study methodology precluded obtaining information of psychiatric diagnoses prior to death. CONCLUSIONS: Tattoos may be possible markers for lethality from both suicide and accidental death in young people, presumably because of shared risk factors such as substance abuse and personality disorder. Affective disorders should receive further, more specific studies. The clinical value of inquiring about tattoos in young people at risk of suicide needs further study. PMID- 10837886 TI - The prolactin response to buspirone in poststroke depression: a preliminary report. AB - BACKGROUND: The issue of whether a serotonergic abnormality is involved in poststroke depression (PSD) was investigated in a sample of poststroke patients. METHODS: The severity of depression was assessed by Hamilton Rating Scale for Depression (HDRS). Buspirone was administered to 16 depressed poststroke (DPS), 10 non-depressed post-stroke (NDPS) patients, and 10 male healthy controls (HCs), to evaluate serotonin (5-HT) function. RESULTS: The prolactin (PRL) response was significantly blunted in DPS patients compared to HCs. There was no significant relationship between the severity of depression and lesion lateralization. Also, no significant differences in buspirone-induced PRL responses were found between DPS patients with right- and left-sided lesions. The severity of depression in DPS patients was not correlated with the time since stroke. CONCLUSIONS: Our results suggest that serotonergic dysfunction may involve in development of poststroke depression. LIMITATIONS: The relatively small sample size and the failure to adequately control for age are major limitations of this study. PMID- 10837887 TI - GM1 enhances the association of neuron-specific MAP2 with actin in MAP2 transfected 3T3 cells. AB - The ganglioside GM1 is a glycosphingolipid which enhances process formation of several neuronal lines and potentiates some growth factor-mediated responses. Previously we have shown that 24 h exposure of Neuro 2a cells to GM1 mobilized the neuron-specific microtubule-associated protein, MAP2, away from microtubule rich areas to areas of neurite sprouting where MAP2 was more closely associated with the subcortical actin network. To examine the role of GM1 in fostering the shift of the association of MAP2 from tubulin to actin, NIH 3T3 cells were co transfected with pHook-1, which expresses a surface antigen, and a construct expressing MAP2. Transfected cells were selected with magnetic beads coated with a hapten that binds to the expressed surface antigen and treated with 150 microg/ml GM1 for 18-24 h. Actin and MAP2 or tubulin and MAP2 were immunolocalized and examined with confocal microscopy. MAP2 was found throughout the cytoplasm as well as associated with actin filaments. As observed previously with Neuro 2a, GM1 treatment of transfected fibroblasts redistributed the MAP2 away from direct association with microtubules to peripheral areas where the association of MAP2 with actin was enhanced. GM1 did not induce neurite-like processes in MAP2-transfected cells. Treatment with cytochalasin B, which is reported to result in process formation, also did not induce neurite-like processes. These studies suggest that GM1's ability to mobilize MAP2 and promote its association with actin is not restricted to neurons. PMID- 10837888 TI - Appearance of central dipsogenic mechanisms induced by dehydration in near-term rat fetus. AB - Cellular dehydration of central osmoreceptors evokes an integration of behavioral (i.e. drinking) and endocrinologic (i.e. arginine vasopressin secretion) responses to maintain body fluid balance. These osmoregulatory mechanisms have been intensely investigated in adult models. However, there has been limited research of the fetal development of neural mechanisms regulating responses to dehydration. Although behavioral and neuroendocrine responses to dehydration have been demonstrated in utero in precocial species (e.g. ovine), there has been no study to date demonstrating that these responses develop before the neonatal period of altricial species (e.g. rat). This study is the first to use the near term rat fetus to investigate the effects of maternal subcutaneous hypertonic (2 M NaCl) or isotonic (0.15 M NaCl) saline injection on fetal plasma osmolality and brain FOS-immunoreactivity (FOS-ir). Maternal subcutaneous hypertonic saline significantly increased maternal and fetal plasma osmolality to similar levels (328+/-6 and 326+/-6 mosM/kg, respectively). In response to plasma hypertonicity, maternal and fetal brain FOS-ir increased significantly in the regions including the lamina terminalis, and the supraoptic and paraventricular nuclei (SON and PVN) of the hypothalamus. Together, these data indicate that central mechanisms for dipsogenic and arginine vasopressin secretory responses to hypertonicity are present and responsive in the fetal rat brain at near-term gestation. However, differences between fetal and maternal FOS-ir mapping suggest that fetal osmoreceptor development is not yet completed near term. PMID- 10837889 TI - Neonatal chlorpyrifos exposure targets multiple proteins governing the hepatic adenylyl cyclase signaling cascade: implications for neurotoxicity. AB - Chlorpyrifos has been hypothesized to interact with receptors and transduction proteins involved in the production of cyclic AMP, contributing to adverse effects on cell replication and differentiation. We studied the effects of neonatal chlorpyrifos exposure on hepatic adenylyl cyclase (AC) activity, as the liver accumulates the highest concentrations of chlorpyrifos and is the site for generation of its active metabolite, chlorpyrifos oxon. Newborn rats were given 1 mg/kg of chlorpyrifos s.c. on PN1-4. On PN5, 24 h after the last dose, AC catalytic activity was induced as assessed by the response to the direct AC stimulant, Mn(2+). In contrast, AC activation dependent upon interaction of the enzyme with G-proteins (forskolin) did not show any enhancement, suggesting impairment of G-protein function. This conclusion was confirmed by impaired responsiveness to fluoride, which directly activates G-proteins. In addition, the response of AC to hormonal signals was altered in a receptor-selective manner, with an enhanced response to glucagon but not to the beta-adrenoceptor agonist, isoproterenol. The effects of chlorpyrifos on AC signaling displayed a critical developmental period of vulnerability, as treatment of older rats (PN11-14) failed to cause substantial induction of AC or interference with G-protein signaling, although it did still enhance the glucagon response. In all cases, the effects of chlorpyrifos disappeared within a few days of discontinuing treatment. These results stand in contrast to the delayed deterioration of AC signaling seen in the brain after the same chlorpyrifos treatment. The temporal and organ selectivity of chlorpyrifos' effects on the AC cascade suggest that disruption of membrane signaling occurs consequent to selective effects on cell development, rather than representing a direct interaction between chlorpyrifos and signaling proteins. PMID- 10837890 TI - The development of the emetic reflex in the house musk shrew, Suncus murinus. AB - The emetic (retching and vomiting) reflex is an important component of the body's defence system against accidentally ingested toxins and emesis is also a common symptom of disease and a side-effect of a number of pharmacological therapies. The development of the reflex has been the subject of few systematic studies. The aim of this study was to characterise the development of the emetic reflex in Suncus murinus (the house musk shrew) using emetic stimuli acting via three different afferent pathways: motion via the vestibular system, pyrogallol via abdominal vagal afferents and resiniferatoxin (a capsaicin analog) via the brainstem. The emetic reflex was not present to any stimulus prior to postnatal day 10 but the onset of the response to motion lagged behind that to the other stimuli in not being present until postnatal day 15. Body weight was not a determinant of the presence of the reflex. It is proposed that the delayed presence of the emetic reflex in Suncus makes it an ideal species in which to investigate factors regulating its development. PMID- 10837891 TI - The course of later generated axons in the developing optic nerve of the chick embryo. A morphometric electron microscopic study. AB - The topographic position of growth cones (GCs) shows the course of ingrowing axons within the optic nerve and allows to draw conclusions with respect to the fiber order in this pathway. Therefore, the topographic distribution and frequency of GCs as well as the proximal and distal axon shaft segments were studied within cross-sections of the distal, middle, and prechiasmatic part of the nerve of 3-8-day-old embryos using electron microscopy. The ingrowth of GCs was not confined to a particular region. Initially, GCs were found near the ventral periphery. With increasing age, simultaneous ingrowth occurred within an area that expanded dorsally. In parallel, GCs also occurred in dorsal regions and eventually in the dorsal periphery. GCs intermingled everywhere with more mature axon profiles. However, youngest profiles predominated ventrally, oldest dorsally. Hence, maturity increased from ventral to dorsal. This indicated that the time of arrival of axons and the topographic position in the cross-section correlated significantly. It is concluded that axons are chronotopically organized, but in a probabilistic sense. The predominant ingrowth of axons in the ventral part may be associated largely with the first wave of neurogenesis of retinal ganglion cells. The ingrowth in dorsal regions of the cross section may be related to later generated axons that enter the nerve following older axons of the same retinal sector as well as axons of neighboring ganglion cells which continue to leave the mitotic cycle while the front of neurogenesis has spread into the periphery. PMID- 10837892 TI - Neonatal capsaicin treatment prevents the normal postnatal withdrawal of A fibres from lamina II without affecting fos responses to innocuous peripheral stimulation. AB - The development of spinal cord sensory pathways has been investigated in postnatal day (P) 21 rat pups following neonatal capsaicin treatment. Capsaicin induced destruction of C fibres was confirmed by 62% loss of Isolectin B4 (IB4) binding and an 86% loss of calcitonin gene-related peptide (CGRP)-immunoreactive small diameter dorsal root ganglion cells. Neonatal capsaicin treatment prevented the normal withdrawal of choleragenoid-horseradish peroxidase (B-HRP)-labelled A fibres from lamina II (substantia gelatinosa) to deeper laminae postnatally. A fibre terminals projected more dorsally, extending into 43% of lamina II compared to vehicle-treated littermates. A small cell loss in, and/or shrinkage of, substantia gelatinosa cannot account for this. These support the concept of a competitive interaction between A and C fibre afferents to establish final terminal fields. However the continued exuberant A fibre termination in capsaicin treated rats did not lead to continued c-fos induction in the superficial dorsal horn by innocuous stimulation. In normal development, exuberant A fibre terminals coincide with c-fos activation in lamina II by innocuous skin stimulation [23]. Despite the continued presence of exuberant A fibre terminals, c-fos was not induced by innocuous peripheral stimulation in P21 capsaicin-treated rats implying that these superficial terminals do not activate lamina II neurons in the same way as in the neonate. PMID- 10837893 TI - Developmental changes in mitochondrial activity and energy metabolism in fetal and neonatal rat brain. AB - Experiments were undertaken to investigate mitochondrial activity and energy metabolism in the developing rat brain from the late fetal stage to the neonatal stage. Samples of cerebral cortical tissue were obtained from fetuses at 14, 16, 18, and 20 days of gestation, and from pups at 1 h, 1 day and 7 days after birth. Mitochondrial respiration was measured polarographically using homogenates. Fetal and neonatal brains were frozen in situ and fluorometric enzymatic techniques were used for the analysis of ATP, ADP, AMP, and lactate. In the fetal brain, there was a gradual increase in stimulated (+ADP) and uncoupled respiratory rates using glutamate and malate as substrates, from 14 days to 20 days of gestation, together with a moderate increase in ATP concentration and in the sum total of adenine nucleotides, and a significant decrease in lactate. Since non-stimulated (-ADP) respiratory rates did not change with increasing gestational age, the respiratory control ratio appeared to increase over the same period. An increase in mitochondrial activity was more pronounced immediately after birth, together with a marked increase in ATP concentration and in the sum total of adenine nucleotides. The highest rate of mitochondrial respiration was observed in 1-hour old pups. These results indicate that, in the rat brain, there is maturation of oxidative metabolism in mitochondria that is initiated in late gestation. Acceleration in mitochondrial respiration occurs immediately after birth in order to maintain high-energy phosphate levels, and this may be crucial for the successful outcome of the newborn. PMID- 10837894 TI - Overexpression of mSim2 gene in the zona limitans of the diencephalon of segmental trisomy 16 Ts1Cje fetuses, a mouse model for trisomy 21: a novel whole mount based RNA hybridization study. AB - Trisomy 21 (Down syndrome) is the most common chromosomal abnormality associated with mental retardation in humans. Sim2, a human homologue of Drosophila sim gene, which acts as a master regulator of the early development of the fly central nervous system midline, is located on chromosome 21, in the Down syndrome critical region, and might therefore be involved in the pathogenesis of some of the morphological features and brain anomalies observed in Down syndrome. We report here the detailed expression pattern of murine mSim2 gene in Ts1Cje mice fetuses, a segmental trisomy 16 mouse model for trisomy 21, and its overexpression in the zona limitans of the diencephalon using a new quantitative method based on the whole-mount RNA hybridization technique. PMID- 10837895 TI - Effects of nitric oxide release in an area of the chick forebrain which is essential for early learning. AB - Extracellular recording techniques were used to study the effects of the nitric oxide releasing agents diethylamine-NO (DEA-NO) and S-nitroso-N-acetyl penicillamine (SNAP) on synaptic transmission in the intermediate and medial part of the hyperstriatum ventrale (IMHV), a part of the domestic chick forebrain that is essential for some forms of early learning. The field response evoked by local electrical stimulation was recorded in the IMHV in an in vitro slice preparation. DEA-NO (100-200 mgr) significantly depressed the field response in a concentration dependent and reversible manner. However, the depression produced by perfusion with 400 mgr DEA-NO, was not reversed following washout of the drug. With 400 mgr DEA-NO, NO reaches a maximum concentration of 10 mgr at 2 min of perfusion, and then declines slowly. SNAP (400 mgr) produced an effect similar to 400 mgr DEA-NO. Neither the immediate nor the longer-term depressive effect of NO is mediated by activation of guanylyl cyclase because in the presence of both low and high doses of ODQ, a potent and selective inhibitor of NO-stimulated guanylyl cyclase, NO produced the same depression of the field response. There is evidence however that the IMHV possesses c-GMP responsive elements since direct perfusion of 8-Br-cGMP (1 mM) produced a long-term but not an immediate depression. The long-term depression produced by 400 mgr DEA-NO was eliminated in the presence of either a selective adenosine A(1) receptor antagonist or an ADP ribosyltransferase inhibitor. It was also possible to prevent the long-term effect in the presence of tetraethyl ammonium a K(+)-channel blocker. These results suggest that the NO may be acting presynaptically in a synergistic fashion with the adenosine A(1) receptor to depress transmitter release. PMID- 10837896 TI - Regulation of glucose transporters during development of the retinal pigment epithelium. AB - The retinal pigment epithelium (RPE) separates the outer retina from its blood supply. To satisfy the retina's large requirement for glucose, the RPE expresses high levels of glucose transporters. In most rat cells, the transporter GLUT3 provides a basal level of transport, but the expression of GLUT1 can be regulated. The opposite is true in chicken (P. Wagstaff, H.Y. Kang, D. Mylott, P.J. Robbins, M.K. White, Characterization of the avian GLUT1 glucose transporter: differential regulation of GLUT1 and GLUT3 in chicken embryo fibroblasts, Mol. Biol. Cell 6 (1995) 1575-1589). We examined chick RPE to determine which isoform is regulated during development, and if the neural retina regulates GLUT expression. By RT-PCR, RPE expressed GLUT1 and GLUT3, but not GLUT2. Only the level of GLUT1 increased between E5 and E18. A corresponding increase in GLUT1 protein was observed by immunoblotting. Most of the increase occurred between E14 and E18, which corresponds to the late stage of tight junction development. A culture model of development was used to examine the intermediate phase, which extends from E7 to E14. While medium conditioned by the neural retina decreased paracellular diffusion across the tight junctions, it increased diffusion through the glucose transporters. Unlike mammals, chick upregulates different isoforms in quiescent RPE and proliferating fibroblasts. Further, the upregulation of glucose transport is coordinated with the development of tight junctions in the blood-retinal barrier. PMID- 10837897 TI - Ethanol-induced alterations in the expression of neurotrophic factors in the developing rat central nervous system. AB - Neonatal rats were exposed to ethanol throughout gestation, or during the early postnatal period (postnatal days 4-10 (P4-10)), and enzyme-linked immunoabsorbent assays were subsequently conducted in order to assess nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3) protein content in hippocampus, septum, cortex/striatum and cerebellum. These determinations revealed that following prenatal ethanol treatment, there were significant ethanol-induced increases in NGF in P1 cortex/striatum, but no changes in any of the three neurotrophic factors (NTFs) in the other brain regions. Cortex/striatal NGF protein returned to control levels by P10. Following early postnatal exposure, BDNF was elevated in hippocampus and cortex/striatum (assessed on P10), and NGF was also enhanced in cortex/striatum at this age. Hippocampal and cortex/striatal BDNF returned to control levels by P21, but cortex/striatal NGF levels remained enhanced at this age. This NTF did not differ in ethanol and control animals by P60, however. The possible significance of elevated levels of NTFs as a function of ethanol exposure is discussed, and it is speculated that while such alterations could play a protective role, increases in these substances during critical developmental periods could also prove to be deleterious, and could even contribute to certain of the neuropathologies which have been observed following developmental ethanol exposure. PMID- 10837898 TI - GAP-43 N-terminal translocation signal targets beta-galactosidase to developing axons in a pan-neuronal transgenic mouse line. AB - Tracing neural connectivity is important in understanding the intricacy of the nervous system as this represents the functional unit throughout the system. Here, we provide evidence that beta-galactosidase (beta-gal) linked to the N terminal axonal translocation signal of GAP-43 provides a reproducible and versatile reporter system for analyzing the developing nervous system in vivo. When expressed by the GAP-43 promoter in transgenic mice, the fusion protein is detected equally within the developing axons of the peripheral and the central nervous systems, directly reflecting the promoter activity. PMID- 10837899 TI - Paradoxical increase of tyrosine hydroxylase-immunoreactive retinopetal fibers in the weaver mouse. AB - Weaver mice undergo apoptosis of the granule cell precursors of the cerebellum and nonapoptotic death of mesencephalic dopaminergic cells during post-natal development. In contrast, the number of retinal dopaminergic cells was transiently increased in weaver compared to control mice [C. Savy, E. Martin Martinelli, A. Simon, C. Duyckaerts, C. Verney, C. Adelbrecht, R. Raisman-Vozari, J. Nguyen-Legros, Altered development of dopaminergic cells in the retina of weaver mice, J. Comp. Neurol. 1999;412:656-668]. While re-examining the retinas, we observed, in the nerve fiber layer, retinopetal tyrosine hydroxylase immunoreactive fibers, which were dramatically increased in number throughout development and adulthood in the weaver compared to control mice. PMID- 10837900 TI - Prenatal morphine exposure alters susceptibility to bicuculline seizures in a sex and age-specific manner. AB - Bicuculline was used to investigate seizure susceptibility in pre- and peripubertal male and female rats exposed prenatally to morphine. Morphine exposed males showed increased seizure susceptibility at prepubertal and decreased susceptibility at peripubertal ages. There was no difference in seizure susceptibility in morphine-exposed females at either age. Therefore, the present data suggest that males are more vulnerable than females to morphine-induced insults during prenatal brain development. PMID- 10837902 TI - Computers in diabetes - an introduction PMID- 10837901 TI - Evaluation of L-DOPA biotransformation during repeated L-DOPA infusion into the striatum in freely-moving young and old rats. AB - The aim of this study was to assess changes in L-3, 4-dihydroxyphenylalanine (L DOPA) biotransformation in response to two-pulse infusion of L-DOPA into the striatum of freely-moving young (3-4 month) and old (21-26 month) male Wistar rats. In addition, the effects of L-DOPA infusion on the vesicular dopamine (DA) store in young rats were also studied. Both L-DOPA-induced DA overflow and uptake of the perfused L-DOPA by the striatum were used to study L-DOPA biotransformation during microdialysis. High potassium-induced DA depletion was performed to assess the dynamics of the vesicular DA store following L-DOPA infusion. Concentric microdialysis probes were stereotaxically implanted in the lateral striatum of rats of both age groups and microdialysis was begun 24 h later. All rats received 2x20 min infusions of 3 mgr L-DOPA separated by an interval of 60 min. In the striatum of both groups, L-DOPA-induced DA overflow and uptake of exogenous L-DOPA were both significantly enhanced during the second infusion compared to the first. In young rats, when a 20-min infusion of 3 mgr L DOPA was given between 2x20 min infusions of 100 mM potassium, no increased DA release was seen at the second high potassium challenge compared with the first. Our results suggest that the enhancement of DA overflow induced by the second L DOPA infusion is, at least partially, due to an increase in L-DOPA biotransformation, and not simply to an enlarged DA pool. In contrast to the in vitro results, our own in vivo results show that L-DOPA utilization in the aging striatum does not deteriorate with age. PMID- 10837903 TI - The integration of digital camera derived images with a computer based diabetes register for use in retinal screening. AB - Exeter district provides a retinal screening service based on a mobile non mydriatic camera operated by a dedicated retinal screener visiting general practices on a 2-yearly cycle. Digital attachments to eye cameras can now provide a cost effective alternative to the use of film in population based eye screening programmes. Whilst the manufacturers of digital cameras provide a database for the storage of pictures, the images do not as yet interface readily with the rest of the patient's computer held data or allow for a sophisticated grading, reporting and administration system. The system described is a development of the Exeter diabetes register (EXSYST) which can import digitally derived pictures from either Ris-Lite TM and Imagenet TM camera systems or scanned Polaroids Pictures can be reported by the screener, checked by a consultant ophthalmologist via the hospital network, and a report, consisting of colour pictures, map of relevant pathology and referral recommendations produced. This concise report can be hard copied inexpensively on a high resolution ink-jet printer to be returned to the patient's general practitioner. Eye images remain available within the hospital diabetes centre computer network to facilitate shared care. This integrated system would form an ideal platform for the addition of computer based pathology recognition and total paperless transmission when suitable links to GP surgeries become available. PMID- 10837904 TI - Screening for diabetic retinopathy using computer based image analysis and statistical classification. AB - Diabetic retinopathy is one of the most common causes of blindness in Europe. However, efficient therapies do exist. An accurate and early diagnosis and correct application of treatment can prevent blindness in more than 50% of all cases. Digital imaging is becoming available as a means of screening for diabetic retinopathy. As well as providing a high quality permanent record of the retinal appearance, which can be used for monitoring of progression or response to treatment, and which can be reviewed by an ophthalmologist, digital images have the potential to be processed by automatic analysis systems. We have described the preliminary development of a tool to provide automatic analysis of digital images taken as part of routine monitoring of diabetic retinopathy in our clinic. Various statistical classifiers, a Bayesian, a Mahalanobis, and a KNN classifier were tested. The system was tested on 134 retinal images. The Mahalanobis classifier had the best results: microaneurysms, haemorrhages, exudates, and cotton wool spots were detected with a sensitivity of 69, 83, 99, and 80%, respectively. PMID- 10837905 TI - Northwest Herts diabetic management system. AB - The Diabetic Management System was set up in Hemel Hempstead within the Dacorum district (population 119515 with 2176 registered patients with diabetes). The objective was to create a clinical management system for all patients with diabetes in a district using a computerised clinical base. To achieve this a database of agreed clinical information was collected from all general practitioners within the district. From the computerised data we can identify those patients who do not have data recorded and can recall them for review. Consultant advice can be given on patients with special or multiple risks identified from interrogation of the database. The outcome has been a higher percentage of patients with diabetes receiving clinical review and achieving optimal clinical targets within the reviews. We already have recorded data on 85% for retinal screening and 75% for comprehensive annual reviews. Conclusions drawn from the Dacorum Diabetic Management System are that effective diabetic care to a population can be provided by a combination of hospital and primary care. Primary care needs to be supported by adequate consultant advice with continued monitoring of the whole population to ensure standards are met. This system achieves this without increased hospital referrals. PMID- 10837906 TI - Computer-aided learning for the education of patients and family practice professionals in the personal care of diabetes. AB - Diabetes Mellitus is approaching pandemic proportions across the globe. It is a disproportionately expensive condition, accounting for 5-9% of annual NHS expenditure. Family practices often play a huge role in the care of diabetic patients. Many GPs elect to play a larger role in diabetes care, but the increasing burden on the multidisciplinary secondary care team means that some of the burden has to fall to family practitioners. In order to provide a high standard of care, the practitioner requires access to continuing education regarding diabetes care. The value of patient education is undisputed. In light of this situation a computer-aided learning (CAL) system is being developed for the education of both patients and practitioners concerning diabetes and its care. The proposed system takes a two pronged approach, being aimed at both patient and practitioner. This interactive system employs multimedia technology to teach practical skills and promote and consolidate theoretical understanding. It is hoped this system will improve patient self-care, and in the long-term reduce the incidence of diabetic complications and their associated costs. PMID- 10837907 TI - Diabetic patients management exploiting case-based reasoning techniques. AB - In this paper we propose a case-based decision support tool, designed to help physicians in 1st type diabetes therapy revision through the intelligent retrieval of data related to past situations (or 'cases') similar to the current one. A case is defined as a set of variable values (or features) collected during a visit. We defined taxonomy of prototypical patients' conditions, or classes, to which each case should belong. For each input case, the system allows the physician to find similar past cases, both from the same patient and from different ones. We have implemented a two-steps procedure; (1) it finds the classes to which the input case could belong; (2) it lists the most similar cases from these classes, through a nearest neighbor technique, and provides some statistics useful for decision taking. The performance of the system has been tested on a data-base of 147 real cases, collected at the Policlinico S. Matteo Hospital of Pavia. The tool is fully integrated in the web-based architecture of the EU funded Telematic management of Insulin Dependent Diabetes Mellitus (T IDDM) project. PMID- 10837908 TI - A model of the endogenous glucose balance incorporating the characteristics of glucose transporters. AB - This paper describes the development and preliminary test of a model of the endogenous glucose balance that incorporates the characteristics of the glucose transporters GLUT1, GLUT3 and GLUT4. In the modeling process the model is parameterized with nine parameters that are subsequently estimated from data in the literature on the hepatic- and endogenous- balances at various combinations of blood glucose and insulin levels. The ability of the resulting endogenous balance to fit blood glucose measured from patients was tested on 20 patients. The fit obtained with this model compared favorably with the fit obtained with the endogenous balance currently incorporated in the DIAS system. PMID- 10837909 TI - Evaluation of DIABNET, a decision support system for therapy planning in gestational diabetes. AB - DIABNET is a knowledge-based system designed to aid doctors with therapy planning in gestational diabetes. The system core is a qualitative model, implemented by a Causal Probabilistic Network, that is able to detect the insulin effectiveness on a daily basis. DIABNET analyses monitoring data and proposes quantitative changes in insulin therapy and qualitative diet modifications. This paper proposes an evaluation methodology to assess the system performance when working in a real scenario. The methodology manages the absence of a gold standard and includes: a subjective analysis based on questionnaires and an objective analysis based on a quantitative comparison of the system's and experts' proposals. The paper also shows the results of two experiments in which expert diabetologists evaluated the therapeutical advice provided by DIABNET during the follow up of 9 patients with gestational diabetes. DIABNET detected the need of a therapy modification in 92% of the cases showing its appropriateness for automatic alarm generation. Around 80% of the proposals were accepted by experts. The evaluation results are encouraging and allow characterisation of the system's performance when proposing therapy modifications. Evaluation in its turn helps to refine the knowledge managed by DIABNET and enables us to look towards the further clinical use of DIABNET as a decision tool in gestational diabetes integrated in a telemedicine service. PMID- 10837910 TI - Models of subcutaneous insulin kinetics. A critical review. AB - Subcutaneous insulin kinetics is a complex process whose quantitation is needed for a reliable glycemic control in the conventional therapy of insulin-dependent diabetes. The major difficulties in modeling include accounting for the distribution in the subcutaneous depot and transport to plasma. A single model describing in detail the various processes for all the commercially available insulin preparations is not available. Several models however have been proposed which vary in the degree of complexity. Virtually all of them handle the regular insulin preparation while a few handle the intermediate acting and the novel insulin analogues. In this paper we critically review these models. PMID- 10837911 TI - A novel kinase, AATYK induces and promotes neuronal differentiation in a human neuroblastoma (SH-SY5Y) cell line. AB - Apoptosis Associated Tyrosine Kinase (AATYK), a novel protein recently isolated from differentiating 32D mouse myeloid cells, contains a putative tyrosine kinase domain and several binding motifs for src homology 2 (SH-2) and src homology 3 (SH-3) domain containing proteins. We observed that AATYK is expressed in different regions of the brain. Although it might play a role in normal nervous system development by modulating apoptosis, little is known regarding its function in the brain or its intracellular localization and kinase activity. Recognizing its homology with Insulin like growth factor-I (IGF-I) receptor (IGF IR) and the critical role of IGF-I in neuronal survival, we hypothesized that AATYK plays an important role in neuronal differentiation/apoptosis. To test this hypothesis, we transfected the human adrenergic neuroblastoma (NB):SH-SY5Y cells with AATYK cDNA under a tetracycline-repressible promoter and established stable cell lines that readily express AATYK on removal of tetracycline. AATYK immunoprecipitated from these cell lysates is an active kinase. Indirect immunofluorescent staining of the clones revealed AATYK to be localized in the cytoplasm. By itself, AATYK overexpression for short duration (2-3 days) did not induce differentiation in the stable SH-SY5Y clones. On the other hand, overexpression for longer periods (7-8 days) per se, significantly (P<0.05-0.001) increased the percent of differentiated cells as well as the neurite length. AATYK-induced differentiation was in the same range as the differentiation induced by agents like all-trans retinoic acid (RA), 12-O-Tetradecanoyl phorbol 13-acetate (TPA) and IGF-I. In addition, AATYK significantly promoted the neuronal differentiation induced by these agents. Our results demonstrate for the first time that AATYK is an active, non-receptor, cytosolic kinase which induces neuronal differentiation and also promotes differentiation induced by other agents in the SH-SY5Y cells. PMID- 10837912 TI - N-Methyl-D-aspartate receptor mediated toxicity in nonneuronal cell lines: characterization using fluorescent measures of cell viability and reactive oxygen species production. AB - Cells transfected with specific N-methyl-D-aspartate (NMDA) receptor subtypes undergo cell death that mimics glutamate-induced excitotoxicity pharmacologically. We have further characterized the mechanisms of cell death resulting from NMDA receptor activation in such cells through development of cell counting methods based on co-transfection with green fluorescent protein. When co transfected with NMDA receptors, GFP expression was limited to live cells as indicated by the observation that GFP was only detected in cells which were positive for markers of live cells, and was found in no cells which were trypan blue or propidium iodide positive. Using co-transfection with green fluorescent protein and cell counting of viable cells with a fluorescence activated cells sorter, we confirmed the subunit-specific profile of NMDA receptor-mediated cell death in cells transfected with NMDA receptors. Toxicity was greatest in the NR1A/2A receptor, less in the NR1A/2B receptor, and least in NR1A/2C receptors. Cell death also differed pharmacologically between subunit combinations. Cell death in cells transfected with NR 1A/2A was blocked by amino-phosphonovaleric acid at lower concentrations than in cells transfected with NR 1A/2B. In cells transfected with the NR1A/2A or NR1A/2B combinations but not NR1A/2C, cell death was also associated with production of reactive oxygen species. In addition, removal of the final 400 amino acids of the C-terminal region of NR2A decreased cell death. The use of GFP based cell counting provides a sensitive mechanism for assessing the mechanism of excitotoxicity in transfected cell models. PMID- 10837913 TI - Impairment of brain mitochondrial function by hydrogen peroxide. AB - Hydrogen peroxide, at concentrations comparable to those observed under some pathological conditions, produced a concentration-dependent inhibition of state 3 (ADP-stimulated) and uncoupled mitochondrial respiratory activity. The ADP:O ratio was also substantially reduced. In contrast, the organic peroxide, t butylhydroperoxide at the same concentrations produced no significant changes in respiratory activity. Intramitochondrial glutathione was oxidised to a similar extent in the presence of hydrogen peroxide or t-butylhydroperoxide. Thus, changes in this endogenous antioxidant apparently did not underlie the different responses to these peroxides. The effects of hydrogen peroxide were not altered by deferoxamine indicating that the extramitochondrial generation of hydroxyl radicals was not likely to be involved. However, modifications arising from the generation of hydroxyl radicals within the mitochondria remain a likely contributor to the observed deleterious effects on respiratory function. The inhibitory effects of hydrogen peroxide were greatest when pyruvate plus malate were present as respiratory substrates. Lesser inhibition was seen with glutamate plus malate and no significant inhibitory effects were detected in the presence of succinate. The findings suggest that mitochondrial components involved in pyruvate oxidation were particularly sensitive to the hydrogen peroxide treatment. However, no significant change was seen in activity of either the pyruvate dehydrogenase complex or NADH-ubiquinone oxidoreductase (complex I) when measured directly following treatment of the mitochondria with hydrogen peroxide. PMID- 10837914 TI - Analysis of the 5'-flanking region of the beta-amyloid precursor protein gene that contributes to increased promoter activity in differentiated neuronal cells. AB - To study the transcription control of the beta-amyloid precursor protein (betaAPP) in Alzheimer's disease (AD), we functionally characterized the betaAPP gene promoter in differentiated cells. PC12 cells were first differentiated with nerve growth factor (NGF) and then transient transfection analysis was done with a series of 5'-deletion constructs, that extended as far upstream as -7900 down to +104 base pair (bp) relative to the transcription start site (+1). The truncated regions of the promoter were linked upstream to a reporter gene, chloramphenicol acetyl transferase (CAT). The CAT assay was performed to compare promoter activity of different 5'-flanking and intronic regions of the betaAPP gene. Our results suggest that the longest (-7900/+104) and one of the shortest ( 47/+104) regions possessed significantly higher levels of promoter activity than the promoterless vector in NGF-differentiated PC12 cells. A deletion of about 7600 bp region from the -7900 to +104 construct resulted in 50% loss of original promoter activity. A deletion of all but 47 bp from the -7900 to +104 construct resulted in the loss of 66% (and retention of 34%) promoter activity. The region 3416/+104 bp displayed the strongest promoter activity whereas +1/+104 bp showed the least activity among all deletion constructs studied. The upstream region 5529 to -3416 contains a negative regulatory element and -3416 to -1131 contains a positive regulatory element. The very upstream region, -7900 to -3411, lacks independent functional activity. The 5'-UTR region (+1 to +104) showed minimum activity and the -75 to +104 region constitutes the basic promoter element. The first exon or a large part of the first intron (+99 to +6200) did not display any significant promoter activity. Thus, several positive and negative regulatory elements influence the basal level of betaAPP promoter activity in NGF differentiated PC12 cells. We speculate that any structural alteration(s) due to a specific mutation in these regulatory regions can potentially alter the transcriptional machinery, and that can perhaps affect the level of beta-amyloid protein involved in AD. PMID- 10837915 TI - Molecular cloning and characterisation of GPR74 a novel G-protein coupled receptor closest related to the Y-receptor family. AB - A novel gene product, GPR74, with homology to the seven transmembrane-domain receptor superfamily, has been cloned. GPR74 has been identified from the expressed sequence tags (EST) database. Subsequent PCR amplification of that sequence and screening of a human heart cDNA library led to the isolation of a 1.7-kb cDNA clone encoding a protein of 408 amino acids. GPR74 shows highest amino acid identity (33%) to the human neuropeptide Y-receptor subtype Y2. The human and mouse genes for GPR74 have been isolated and their exon-intron structures determined. In both species the gene consists of four exons spanning around 20 kb with the exon-intron borders being 100% conserved. Northern analysis of various human tissues reveals highest levels of mRNA expression in brain and heart. In situ hybridisation analysis of rat brain tissue confirms this result and identifies the hippocampus and amygdala nuclei as the brain areas with particular high expression of GPR74 mRNA. Fluorescence in situ hybridisation, PCR analysis on a radiation hybrid panel and interspecific mouse backcross mapping have localised the genes to human chromosome 4q21 and mouse chromosome 5. Expression of the human GPR74 cDNA as a GFP-fusion protein in various cell lines reveals the inability of the recombinant receptor protein to reach the cell surface. This is consistent with the lack of NPY specific binding in these cells and suggests that unknown factors are required for a full functional receptor complex. PMID- 10837916 TI - Expression and role of p27(kip1) in neuronal differentiation of embryonal carcinoma cells. AB - We examined the expression and the regulation of p21(waf1) and p27(kip1) cdk inhibitors in P19 mouse embryonal carcinoma (EC) cells following treatment with all-trans retinoic acid (ATRA) to induce neuronal differentiation. The levels of p27 mRNA and protein increased within 24 h of treatment with ATRA, reaching a plateau 4-5 days later prior to neurite formation. In contrast, levels of p21 expression remained low until after neurites were extensively formed. Induction of muscle differentiation from P19 cells by treatment with dimethyl sulfoxide caused only transient increases in p27 levels. In a mutant P19 cell line, RAC65, treatment with ATRA induced neither p27 accumulation nor neuronal differentiation, but p21 mRNA expression increased markedly. In contrast, treatment of RAC65 cells with 9-cis retinoic acid induced both p27 expression and neuronal differentiation. Correlation between p27 expression and neuronal differentiation was also observed in NT2/D1 human EC cells. Luciferase reporter assays showed that p27 promoter activity increased in ATRA-treated cells, consistent with the elevation of p27 mRNA levels. Arrest of neuronal differentiation of P19 cells by okadaic acid resulted in inhibition of p27 expression, whereas p21 mRNA expression was greatly enhanced. Conversely, inhibition of p27 expression by antisense p27 oligonucleotides resulted in blockade of neuronal differentiation. Taken together, these results strongly suggest that the expression of p27 is indispensable for neuronal differentiation of EC cells. PMID- 10837917 TI - Molecular activation of noradrenergic neurons in the rabbit brainstem after coitus. AB - Our previous studies indicate that coitus in female rabbits induces a gonadotropin-releasing hormone (GnRH) surge that is preceded by an increase in hypothalamic norepinephrine (NE) release. The additional findings of an enhanced tyrosine hydroxylase (TH) mRNA expression in the female brainstem after coitus, in addition to the appropriate topographic distribution of TH and dopamine-beta hydroxylase (DBH), lead us to hypothesize that coital signals are relayed to hypothalamic GnRH-secreting neurons via brainstem NE-containing perikarya. Here we analyzed coitally activated areas in the brainstem by in situ hybridization of the oncogene c-fos, as well as the expression of TH mRNA at 0, 30 and 60 min postcoitus using specific 35S-labeled probes for c-fos and TH. To establish the identity of activated brainstem neurons, we immunocytochemically double-labeled cells with specific antibodies against Fos protein and DBH at 90 min postcoitus. Both c-fos and TH mRNAs were present at 0 min (control) in the A1, A2 and A6 brainstem-noradrenergic areas. At 30 min after coitus the expression of both genes significantly increased (P<0.01) in the A1 and A2 areas. By 60 min postcoitus the expression of c-fos mRNA decreased to control levels, while that of TH mRNA remained stimulated. Double-labeling of Fos and DBH indicated that the number of dual-labeled neurons increased (P<0.05) over control levels only in the A1 and A2 areas (not in A6) at 90 min postcoitus. These findings support the hypothesis that coitus activates transcriptional/translational events within brainstem NE neurons that culminate in the release of hypothalamic NE and hence a GnRH surge. PMID- 10837918 TI - Wnt-1 inhibits nerve growth factor-induced differentiation of PC12 cells by preventing the induction of some but not all late-response genes. AB - The vertebrate Wnt-1 proto-oncogene is expressed transiently in embryonic brain and functions in the development of the central nervous system and neural crest. The role of Wnt-1 in neural crest development appears to be to increase the number of certain progenitor cells by preventing their premature differentiation. To study the mechanism by which this transient Wnt-1 expression inhibits differentiation we have constructed PC12 pheochromocytoma cells in which Wnt-1 expression levels were controlled by use of a tetracycline-responsive transactivator. Induction of Wnt-1 expression by tetracycline withdrawal was followed by activation of the Wnt-1 signalling pathway as shown by activation of the Lef-1/Tcf transcription factor. Wnt-1 expression by these cells resulted in reversible inhibition of NGF-induced neurite outgrowth, but it did not adversely affect the maintenance of previously formed NGF-induced neurites. Wnt-1 expression also partially blocked the ability of NGF to decrease the rate of cell multiplication. Wnt-1 decreased the NGF-induced expression of the late-response gene SCG10 but not of the immediate early genes, fos, Nur77 and UPAR (urokinase type plasminogen activator receptor) nor of the late-response genes GAP-43 and collagenase. The Wnt-1 expressing PC12 cells multiplied at a greater rate when they expressed Wnt-1 than they did in the absence of Wnt-1 expression, a result that is consistent with the proposal that Wnt-1 may also act as a mitogen. PMID- 10837919 TI - NMDAR1 isoforms in the rat superior olivary complex and changes after unilateral cochlear ablation. AB - Normal expression and deafness related changes in expression of NMDAR1 isoforms were examined in the rat superior olivary complex (SOC) using in situ hybridization with S35 labeled oligoprobes. Expression was assessed in three SOC nuclei, the lateral and medial superior olives (LSO, MSO) and the medial nucleus of the trapezoid body (MNTB). Silver grain labeling over principal cells of each region was assessed using METAMORPH image analysis system. Counts were made in ipsi- and contralateral sides after unilateral cochlear ablation and in treated and untreated animals. In the normal SOC, NMDAR1a expression was higher than 1b and 1-2 expression was followed by 1-4 and 1-1, with 1-3 below the level for detection. The levels and ratio were comparable in LSO, MSO and MNTB. Five days after cochlear ablation 1a, 1-1, 1-2 and 1-4 showed significant decreases in the ipsilateral LSO and 1-a and 1-2 showed significant decreases in the contralateral MNTB, with no significant changes in the MSO. At 20 days after deafening, no significant changes were seen for any isoform in any nucleus. The transient deafness-induced decreases in expression of NMDAR1 isoforms correlate with loss of excitation. PMID- 10837920 TI - Sequential increase in Egr-1 and AP-1 DNA binding activity in the dentate gyrus following the induction of long-term potentiation. AB - Establishment of long-term potentiation (LTP) at perforant path synapses is highly correlated with increased expression of Egr and AP-1 transcription factors in rat dentate gyrus granule cells. We have investigated whether increased transcription factor levels are reflected in increased transcription factor activity by assessing Egr and AP-1 DNA binding activity using gel shift assays. LTP produced an increase in binding to the Egr element, which was NMDA receptor dependent and correlated closely with our previously reported increase in Egr-1 (zif/268) protein levels. Supershift analysis confirmed involvement of Egr-1, but not Egr-2 in the DNA binding activity. AP-1 DNA binding was also rapidly elevated in parallel with protein levels, however, the peak increase in activity was delayed until 4 h, a time point when we have previously shown that only jun-D protein was elevated. These data indicate that binding of Egr-1 and AP-1 to their response elements is increased in two phases. This may result in activation of distinct banks of target genes which contribute to the establishment of persistent LTP. PMID- 10837921 TI - Identification of morphine in the rat adrenal gland. AB - Morphine was identified in rat adrenal extracts by reverse-phase HPLC, following liquid and solid extraction. All experiments were carefully performed to prevent exogenous morphine contamination. The morphine extracted from adrenal tissue (105.31 ng/g of wet adrenal gland) was identical to that of a morphine internal standard. The morphine corresponding HPLC fractions were further analyzed by gas chromatography-mass spectrometry and found to be identical to synthetic morphine. The study conclusively proves morphine is endogenous to the rat adrenal gland. PMID- 10837922 TI - N-myc and c-myc expression in Alzheimer disease, Huntington disease and Parkinson disease. AB - The present study examines N-myc and c-myc protein expression with Western blotting and single and double-labeling immunohistochemistry in the hippocampus in Alzheimer disease (AD), the striatum in Huntington disease (HD) and the substantia nigra in Parkinson disease (PD). No modifications in the N-myc and c myc expression are found in hippocampal neurons in AD, striatal neurons in HD, and pigmented neurons of the substantia nigra in PD. Yet punctate synaptic-like N myc immunoreactivity, matching enhanced synaptophysin expression, occurs in diffuse plaques, but not in dystrophic neurites of neuritic plaques. In contrast, c-myc immunoreactivity is found in dystrophic neurites, but not in aberrant sproutings of neuritic plaques, as shown by double-labeling immunohistochemistry to c-myc and phosphorylated tau or phosphorylated neurofilament epitopes, and to c-myc and GAP-43, respectively. Strong N-myc and c-myc are observed in reactive astrocytes in AD, HD and PD, as revealed by double-labeling with N-myc or c-myc and GFAP. Finally, no relationship is found between nuclear DNA fragmentation and increased N-myc or c-myc expression in individual cells. These results demonstrate that neuron death in AD, HD and PD is not associated with modifications in the steady-state expression of N-myc and c-myc in individual neurons, and that neurofibrillary degeneration and Lewy body formation are not accompanied by increased immunoreactivity to these transcription factors. Increased N-myc and c-myc expression in reactive astrocytes probably plays a role in reactive astrocytosis in human neurodegenerative disorders. PMID- 10837923 TI - Differential expression of the estrogen receptor-related receptor gamma in the mouse brain. AB - To elucidate estrogen functions, the expression of the estrogen receptor-related receptor ERRgamma, a novel orphan nuclear receptor regulating transcription via estrogen responsive elements, has been localized by in situ hybridization in adult murine brain. ERRgamma transcripts were abundantly present in the isocortex, the olfactory system, cranial nerve nuclei and major parts of the coordination centers, e.g. reticular formation and major parts of the extrapyramidal motor systems. In addition, ERRgamma expression was detected in trigeminal ganglion neurons. ERRgamma distribution was clearly distinguished from that described for ERRalpha, for ERRbeta, and for estrogen receptors (ER) pointing at functional differences between ERRgamma and these receptors. PMID- 10837924 TI - The human dopamine D2(Longer) receptor has a high-affinity state and inhibits adenylyl cyclase. AB - Brain dopamine D2 receptors are the main targets for antipsychotic and anti Parkinsonian drugs. The dopamine D2 receptor has three forms, D2(Short), D2(Long) and D2(Longer). D2(Longer) is a newly found splice variant which contains two additional amino acids (valine and glutamine) in the third cytoplasmic loop of the receptor. To determine whether D2(Longer) was functional, the cDNA was transfected into CHO cells. D2(Longer) revealed a high-affinity state for dopamine ( approximately 1.5 nM), and mediated dopamine-inhibited adenylyl cyclase. PMID- 10837925 TI - Murine aspartoacylase: cloning, expression and comparison with the human enzyme. AB - Canavan disease is caused by mutations in aspartoacylase, the enzyme that degrades N-acetylaspartate (NAA) into acetate and aspartate. Murine aspartoacylase (mASPA) was cloned using sequence information from mouse expressed sequence tags homologous to the human cDNA. The open reading frame was cloned into a thioredoxin fusion vector, overexpressed in bacteria, and the protein was purified using affinity chromatography to near homogeneity. Recombinant human ASPA (hASPA) was prepared by a similar method. Both recombinant enzymes were highly specific to NAA, with about 10% of the NAA activity toward N acetylasparagine. More interestingly, the product of N-acetylasparagine was aspartate but not asparagine, indicating that ASPA catalyzed deacetylation as well as hydrolysis of the beta acid amide. Our success in preparing the recombinant ASPA in high purity should permit multiple lines of investigations to understand the pathogenic mechanisms of Canavan disease and the functional roles of NAA. PMID- 10837926 TI - Detection of the interleukin 18 family in rat brain by RT-PCR. AB - Interleukin 18, an inflammatory cytokine, mediates its effects by interaction with its receptor complex, consisting of the IL-18 receptor (IL-18R) and receptor accessory protein (AcPL). A functional inhibitor of IL-18, the IL-18 binding protein (IL-18BP), has been identified recently. This study reports the detection of IL-18, IL-18R, AcPL and IL-18BP mRNA expression in the brain of normal adult rats using RT-PCR. PMID- 10837927 TI - Early phase collagen synthesis in lungs of rats exposed to bleomycin. AB - Skin wound healing exhibits type III collagen synthesis occurring transiently as early as 10 h after injury, with subsequent synthesis of type I to form a scar. We hypothesized that similar collagen type switching also occurred in the bleomycin model of lung fibrosis in the rat. We could measure elevated lung collagen synthesis rates as early as 4 days after administration of bleomycin. Collagen type I:III ratios in whole lung remained constant for the first 7 days at the control level of 2:1, then increased to as high as 5:1 at day 21. Procollagen mRNA content, expressed as a ratio of type I:III mRNAs, was consistent with the protein synthesis data and the observed ratio of collagen types being made by the lungs at the various time points evaluated. We conclude that a transient increase in type III relative to type I collagen does not occur in the bleomycin rat lung model. Therefore, the sequence of type-specific collagen expression and deposition in the skin wound healing model is not entirely analogous to this widely used animal model of pulmonary fibrosis. PMID- 10837928 TI - Gonadotropin-releasing hormone (GnRH) partially reverses the inhibitory effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin on ovulation in the immature gonadotropin treated rat. AB - Several studies have shown that 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) has inhibitory effects on ovulation. This action may be the result of either direct effect(s) of TCDD on ovarian function or via altered secretion of pituitary luteinizing hormone (LH) and follicle stimulating hormone (FSH) which regulate ovarian follicular development and ovulation. To further evaluate the effects of TCDD on pituitary gonadotropins and their regulation, the potential role of gonadotropin-releasing hormone (GnRH) was investigated in the current study. Immature (23-day-old) female Sprague-Dawley rats were dosed with TCDD (32 microg/kg) in corn oil or vehicle alone. Equine chorionic gonadotropin (eCG) was injected subcutaneously (5 IU, sc) 24 h later to induce follicular development. Immediately prior to the expected time of the LH/FSH surges, 54 h after eCG injection, half of TCDD- or corn oil-treated rats were injected with GnRH (2 microg/rat, sc). Blood and ovaries were collected at 54, 56, 58, 60 and 72 h after eCG. Serum concentrations of 17beta-estradiol (E(2)), progesterone (P(4)), LH, and FSH were determined by radioimmunoassay. An indication of ovulation rate was assessed at 72 h after injection of eCG by irrigating the ova from oviducts. TCDD reduced the number of ova in the oviducts by 70-80% (2-3 ova/rat) and this was confirmed by the number of corpora lutea. GnRH partially restored ovulation (6-7 ova/rat) in TCDD-treated rats without reversing its effect on ovarian weight reduction. In controls, the LH and FSH surges at 58 h after eCG were significantly reduced at that time in TCDD-treated rats. However, in rats treated with TCDD and GnRH, a huge LH/FSH surges occurred at 56 h after eCG injection. GnRH alone enhanced E(2) and P(4) serum levels at 56-58 h after eCG injection. In rats treated with both TCDD and GnRH, E(2) secretion was significantly lower at 58, 60, and 72 h when compared with GnRH alone, whereas serum P(4) was only decreased at 72 h after eCG injection. The results indicate that exogenous GnRH induces LH and FSH surges in TCDD-treated rats, but only partially restores the inhibitory effects of TCDD on ovulation. PMID- 10837929 TI - Simultaneous administration of diethylphthalate and ethyl alcohol and its toxicity in male Sprague-Dawley rats. AB - Phthalate esters have been implicated as xenoestrogens. One among them is di ethylphthalate (DEP), which is used as plasticizer, detergent base, and binder in incense sticks and after-shave lotions. DEP is one of the contaminants of freshwater and marine ecosystems. Incense stick workers are occupationally exposed to DEP and some workers are chronic alcoholics. Therefore, a study was undertaken to evaluate the interactive toxicity of DEP with ethyl alcohol (EtOH) in young male Sprague-Dawley rats. The rats were given 50 ppm DEP (w/v), 5% EtOH (v/v) and a combined dose of 50 ppm DEP (w/v)+EtOH (5% v/v) in water ad libitum for a period of 120 days and were maintained on normal diet. Control animals received normal diet and plain water. During the treatment rats were weighed every week and water consumption per day was measured. After the completion of treatment, liver weight/body weight, liver weight, body weight, serum enzymes and other biochemical parameters were assessed. It was found that there was no significant change observed in body weight, liver weight, liver weight/body weight and water consumption. It was observed that there was a significant decrease in liver aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels in EtOH, DEP and EtOH+DEP treated rats in the order of EtOH>DEP>EtOH+DEP as compared with control. Serum AST, ALT, acid phosphatase (ACP), alkaline phosphatase (ALP), succinate dehydrogenase (SDH) and liver ACP showed significant increase in DEP and EtOH+DEP treated rats in the order of DEP>EtOH+DEP as compared with control and EtOH treated rats. On the contrary, there was no significant change in liver ALP levels in treated rats. There was significant increase in liver SDH, glycogen, total triglyceride, total cholesterol and lipid peroxidation in DEP and EtOH+DEP treated rats, but no significant changes in the serum SDH, glucose and total triglyceride levels. Serum total cholesterol levels in DEP and EtOH+DEP treated rats were significantly high as compared to control and EtOH treated rats. These results show that there is no interaction of DEP with EtOH but DEP alone leads to severe impairment of lipid metabolism coupled with toxic injury to the liver as evident from significantly altered lipid and enzyme levels in the liver and serum. Long term simultaneous exposure to DEP and EtOH may have severe implications for humans who are occupationally exposed to these two xenobiotics. PMID- 10837930 TI - Kinetic analysis of the toxicological effect of tacrine (Cognex) on human retinal acetylcholinesterase activity. AB - For the first time, kinetic parameters of the effect of tacrine, an anti cholinesterase inhibitor of therapeutic potential in Alzheimer's disease has been studied on human retinal acetyl-cholinesterase (AChE). Tacrine inhibited the AChE activity in a concentration dependent manner, the IC(50) being about 45 nM. The Michaelis-Menten constant (K(m)) for the hydrolysis of acetylthiocholine iodide was found to be 0.120 mM and this value was increased by 4-52.8% in the presence of tacrine. V(max) was observed to be 2.23 micromol/h per mg protein for the control system, while it was decreased by 14.73-56.25% in the tacrine treated systems. Dixon as well as Lineweaver-Burk plots and their secondary replots indicated that the nature of the inhibition was of the mixed type, i. e. a combination of competitive and noncompetitive inhibition. The values of K(i) and K(I) were estimated to be as 37.76 and 64.36 nM, respectively. PMID- 10837931 TI - Implications of oxidative stress and inflammatory process in the cytotoxicity of capsaicin in human endothelial cells: lack of DNA strand breakage. AB - Capsaicin, a natural product of Capsicum species is known to induce excitation of nociceptive terminals involved in pain perception. Nevertheless, it is utilized by topical application in humans, giving rise to blood capsaicin concentration up to 10-20 microM. The effect of capsaicin on human endothelial cells ECV 304 has been investigated. The cytotoxicity and inflammatory properties of capsaicin were evaluated by measuring the capsaicin-stimulated release of soluble intercellular adhesion molecule-1 levels (sICAM-1) into the culture medium; production of reactive oxygen species measured by quantification of lipoperoxidation in endothelial cell membranes; and genotoxicity measured using the comet assay and the DNA fragmentation assay. The concentration inhibiting protein synthesis by 50% after 24-h incubation was found to be 175 microM. Capsaicin induced an increase of sICAM-1 release into the culture medium at concentration >/=100 microM. Lipoperoxidation measured by malondialdehyde production increased at capsaicin concentration >/=200 microM. The comet test and DNA fragmentation assay clearly suggested that capsaicin does not induce significant DNA strand breaks within the range of concentrations used. Because the inflammatory reaction and lipid peroxidation may affect cellular functions and lead to cell death, the present data may have important implications for the possible health threats of capsaicin, specially in the case of unreasonable use of capsaicin preparations in pathological situations. PMID- 10837932 TI - Suppression of autophagy by ethionine administration in male rat liver in vivo. AB - The administration of ethionine to male rats decreased the liver adenosine triphosphate (ATP) levels to about 20% of that in control rats and prevented leupeptin-induced intralysosomal accumulation of cytosolic proteins, including lactate dehydrogenase sequestered into lysosomes by autophagy, and leupeptin induced increase of lysosomal density. These actions of ethionine were reversed by the subsequent administration of adenine plus methionine. All these findings show that the administration of ethionine to male rats suppresses the autophagic sequestration of intracellular proteins into lysosomes, probably due to ethionine induced ATP depletion. PMID- 10837933 TI - In vitro aberrant gene expression as the indicator of lead-induced neurotoxicity in U-373MG cells. AB - Lead is an important neurobehavioral toxicant and may interfere with developmental processes in the brain resulting in impairment of its functions. U 373MG, a human glioma cell line, was cultured in Dulbecco's modified Eagles' medium supplemented with either 20 or 10% FBS (fetal bovine serum) to explore the possible indications for lead-induced toxicity. Although lead did not affect cell growth rate in concentrations ranging from 0.01 to 10 microM, it substantially altered gene expression analyzed by reverse-transcription polymerase chain reaction. With 10% FBS culture, lead affected the gene expression in a dose dependent relationship. It enhanced the expression of tumor necrosis factor-alpha (TNF-alpha), but decreased those of interleukin-1beta (IL-1beta), interleukin-6 (IL-6), gamma-aminobutyric acid (GABA) transaminase, and glutamine synthetase. With 20% FBS culture, lead also profoundly increased TNF-alpha and IL-1beta; however, it did not extensively affect the other genes examined above. Thus, the highly sensitive changes of gene expression of these cytokines or metabolic enzymes after treatments with lead acetate evidenced their usefulness as indicators for in vitro measurement of lead-induced neurotoxicity. PMID- 10837934 TI - Tumor radiosensitivity: it's the subpopulations that count. PMID- 10837935 TI - Towards multidimensional radiotherapy (MD-CRT): biological imaging and biological conformality. AB - PURPOSE: The goals of this study were to survey and summarize the advances in imaging that have potential applications in radiation oncology, and to explore the concept of integrating physical and biological conformality in multidimensional conformal radiotherapy (MD-CRT). METHODS AND MATERIALS: The advances in three-dimensional conformal radiotherapy (3D-CRT) have greatly improved the physical conformality of treatment planning and delivery. The development of intensity-modulated radiotherapy (IMRT) has provided the "dose painting" or "dose sculpting" ability to further customize the delivered dose distribution. The improved capabilities of nuclear magnetic resonance imaging and spectroscopy, and of positron emission tomography, are beginning to provide physiological and functional information about the tumor and its surroundings. In addition, molecular imaging promises to reveal tumor biology at the genotype and phenotype level. These developments converge to provide significant opportunities for enhancing the success of radiotherapy. RESULTS: The ability of IMRT to deliver nonuniform dose patterns by design brings to fore the question of how to "dose paint" and "dose sculpt", leading to the suggestion that "biological" images may be of assistance. In contrast to the conventional radiological images that primarily provide anatomical information, biological images reveal metabolic, functional, physiological, genotypic, and phenotypic data. Important for radiotherapy, the new and noninvasive imaging methods may yield three dimensional radiobiological information. Studies are urgently needed to identify genotypes and phenotypes that affect radiosensitivity, and to devise methods to image them noninvasively. Incremental to the concept of gross, clinical, and planning target volumes (GTV, CTV, and PTV), we propose the concept of "biological target volume" (BTV) and hypothesize that BTV can be derived from biological images and that their use may incrementally improve target delineation and dose delivery. We emphasize, however, that much basic research and clinical studies are needed before this potential can be realized. CONCLUSIONS: Whereas IMRT may have initiated the beginning of the end relative to physical conformality in radiotherapy, biological imaging may launch the beginning of a new era of biological conformality. In combination, these approaches constitute MD-CRT that may further improve the efficacy of cancer radiotherapy in the new millennium. PMID- 10837936 TI - A detrimental effect of a combined chemotherapy-radiotherapy approach in children with diffuse intrinsic brain stem gliomas? AB - PURPOSE: To compare the proportion of patients that survive at least 1 year following treatment with hyper-fractionated radiotherapy (HRT) to a dose of 70.2 Gy on Pediatric Oncology Group (POG) study #8495 with that of patients treated with similar radiotherapy plus cisplatinum given by continuous infusion on weeks 1, 3, and 5 of radiotherapy on POG #9239. METHODS AND MATERIALS: The eligibility criteria for the two studies were identical and included age 3 to 21 years, previously untreated tumor involving the brain stem of which two-thirds was in the pons, history less than 6 months, and clinical findings typical for diffuse intrinsic brain stem glioma, including cranial nerve deficits, long tract signs, and ataxia. The outcome of 57 patients who were treated at the 70.2 Gy dose level of POG #8495 between May 1986 and February 1988 was compared with that of 64 patients treated with identical radiotherapy plus cisplatinum on POG #9239 between June 1992 and March 1996. RESULTS: The number of patients accrued to POG #9239 was determined to guarantee that the probability was at least 0.80 of correctly detecting that the 1-year survival rate exceeded that of patients on POG #8495 by 0.2. However, the z value for this test was -1.564, giving a p value of 0.9411. That is, there is almost sufficient evidence to conclude that survival for patients receiving HRT plus cisplatinum on POG #9239 was worse than that for patients receiving the same radiotherapy alone on POG #8495. CONCLUSION: The finding that patients who received cisplatinum given as a radiosensitizing agent concurrent with HRT fared less well than those receiving the same dose of HRT alone was unexpected and is clearly a cause for concern as many current protocols for patients with diffuse intrinsic brain stem gliomas call for use of chemotherapeutic and/or biological agents given concurrent with radiotherapy. PMID- 10837937 TI - Transcranial sonography: integration into target volume definition for glioblastoma multiforme. AB - PURPOSE: Recent studies indicate that transcranial sonography (TCS) reliably displays the extension of malignant brain tumors. The effect of integrating TCS into radiotherapy planning for glioblastoma multiforme (GBM) was investigated herein. METHODS AND MATERIALS: Thirteen patients subtotally resected for GBM underwent TCS during radiotherapy planning and were conventionally treated (54 to 60 Gy). Gross tumor volumes (GTVs) and stereotactic boost planning target volumes (PTVs, 3-mm margin) were created, based on contrast enhancement on computed tomography (CT) only (PTV(CT)) or the combined CT and TCS information (PTV(CT+TCS)). Noncoplonar conformal treatment plans for both PTVs were compared. Tumor progression patterns and preoperative magnetic resonance imaging (MRI) were related to both PTVs. RESULTS: A sufficient temporal bone window for TCS was present in 11 of 13 patients. GTVs as defined by TCS were considerably larger than the respective CT volumes: Of the composite GTV(CT+TCS) (median volume 42 ml), 23%, 13%, and 66% (medians) were covered by the overlap of both methods, CT only and TCS only, respectively. Median sizes of PTV(CT) and PTV(CT+TCS) were 34 and 74 ml, respectively. Addition of TCS to CT information led to a median increase of the volume irradiated within the 80% isodose by 32 ml (median factor 1.51). PTV(CT+TCS) volume was at median 24% of a "conventional" MRI(T2)-based PTV. Of eight progressions analyzed, three and six occurred inside the 80% isodose of the plans for PTV(CT) and for PTV(CT+TCS), respectively. CONCLUSION: Addition of TCS tumor volume to the contrast-enhancing CT volume in postoperative radiotherapy planning for GBM increases the treated volume by a median factor of 1.5. Since a high frequency of marginal recurrences is reported from dose escalation trials of this disease, TCS may complement established methods in PTV definition. PMID- 10837938 TI - Prognostic factors for medulloblastoma. AB - PURPOSE: To evaluate prognostic factors for medulloblastoma. METHODS AND MATERIALS: One hundred and seventy-three consecutive patients with medulloblastoma, treated at King Faisal Specialist Hospital (KFSH) from 1988 1997, were reviewed. Eighty-four percent were children less than 15 years old. From 1988-1994, treatment was at the discretion of the investigator. From 1994 1998, patients entered a single-arm best practice protocol in which, in staged patients, the surgical intent was total resection, standard radiation treatment was defined, and adjuvant chemotherapy was given to a "high-risk" subset. RESULTS: For 150 patients who completed surgical and radiation treatment, the 5 year survival rate was 58%, compared with 0% for 16 patients who were unable to start or complete radiation treatment. For staged patients, the 5-year survival was M0 + M1, 78% and M2 + M3, 21% (p < 0.0001). Other favorable significant prognostic factors were age >14 years and gross cystic/necrotic features in the primary tumor. The size of the primary tumor, the degree of hydrocephalus at diagnosis, the presence of residual tumor in the post-operative CT/MRI, and the functional status of the patient prior to radiation treatment were not significant factors. CONCLUSIONS: Stage M0 + M1 was the most powerful favorable prognostic factor. In Saudi Arabia more patients present with advanced disseminated disease, 41% M2 + M3, than in the West, and this impacts adversely on overall survival. Total resection and standard radiation treatment were not sensitive prognostic factors in a treatment environment in which 78% of patients underwent at least 90% tumor resection and 60% received standard radiation treatment. In order to improve the proportion of patients able to complete radiation treatment, consideration should be given to limiting resection when the attainment of total resection is likely to be morbid, and to delaying rather than omitting radiation treatment in the patient severely compromised postoperatively. PMID- 10837939 TI - The significance of radiotherapy treatment duration in intracranial ependymoma. AB - PURPOSE: To determine if radiation therapy treatment duration has an impact on the outcome of intracranial ependymoma. METHODS AND MATERIALS: From 1965 to 1997, 34 patients with intracranial ependymoma were seen and treated with postoperative radiotherapy (RT). There were 26 male and 8 female patients with a median age of 14 years (range, 18 months to 60 years). Tumor location was infratentorial in 23 (67.6%) and supratentorial in 11 (32.4%). Pathology was low-grade in 25 (73.5%) and high-grade in 9 (26.5%). Two (5.9%) patients had M+ disease at initial diagnosis. Gross total resection was achieved in 12 (35.3%), subtotal resection in 19 (55. 9%), and biopsy alone in 3 (8.8%). Median RT dose to the primary site was 5400 cGy (range, 4500 cGy to 6600 cGy). The entire neuraxis was treated in 14 (41.2%), whole brain in 9 (26.5%), and the primary site alone in 11 (32.4%). Median radiotherapy treatment duration was 43 days (range, 26 to 122 days). Patients treated with craniospinal RT were more likely to have treatment duration of > or =50 days. Median follow-up was 108 months (range, 24 to 252 months). RESULTS: The 5-, 10-, and 15-year overall survival rates were 71.5%, 63.3%, and 55.4%. Local control rates at 5, 10, and 15 years were 61.8%, 57.7%, and 57.7%. For patients with RT treatment duration <50 days, the 5-, 10-, and 15-year overall survivals were 85.5%, 78.9%, and 65.7% whereas for those with treatment duration > or = 50 days, the corresponding rates were 45.5%, 36.4%, and 36.4% (p = 0.01, log-rank test). Local control rate at 5, 10, and 15 years was 70.6% for patients with RT treatment duration <50 days and 45.5%, 36.4%, and 36.4% for patients with RT treatment duration > or =50 days (p = 0.05, log-rank test). Age, gender, tumor location, tumor grade, degree of surgical resection, RT volume, and RT dose (4500-5399 cGy vs. 5400-6600 cGy) were not found to have a statistically significant impact on overall survival, disease-free survival, or local control. The most common site of failure was local, found in 13 (38.2%) patients. Three patients also had spinal failure in addition to a primary site failure. There were no isolated nonprimary site brain or spine failures. CONCLUSIONS: Radiation therapy treatment duration is an important prognostic factor for survival and local control with patients having treatment duration <50 days having a better outcome than those with treatment duration > or =50 days. Because the overwhelming site of failure was local, we recommend local field irradiation in patients with M0 disease. PMID- 10837940 TI - Stereotactic fractionated radiotherapy for chordomas and chondrosarcomas of the skull base. AB - PURPOSE: To investigate the treatment outcome of patients suffering from skull base chordoma or chondrosarcoma after fractionated stereotactic radiotherapy. METHODS AND MATERIALS: We report 45 patients treated for chordoma or chondrosarcoma with postoperative fractionated stereotactic radiotherapy between 1990 and 1997. Patients had CT and MRI for 3D treatment planning performed under stereotactic guidance. Median dose at isocenter was 66.6 Gy for chordomas and 64.9 Gy for chondrosarcomas. MRI imaging was obtained in intervals after therapy to evaluate local relapse. Survival was calculated according to the Kaplan-Meier method. RESULTS: All chondrosarcomas had achieved and maintained local control and recurrence-free status at follow-up of 5 years. Local control rate of chordomas was 82% at 2 years and 50% at 5 years. Survival was 97% at 2 years and 82% at 5 years. At maximum follow-up of 8 years local control and survival rate of chordomas was 40% (82%). Clinically significant late toxicity developed in one patient. CONCLUSIONS: Our results demonstrate the feasibility of fractionated photon beam therapy and its success in the treatment of skull base tumors. Modern 3D treatment techniques provide superior results compared to conventional techniques. The role of high-precision radiotherapy compared to particle beam therapy in the treatment of these tumors is not yet fully clear and further research is needed. PMID- 10837941 TI - Calculation of cranial nerve complication probability for acoustic neuroma radiosurgery. AB - PURPOSE: Estimations of complications from stereotactic radiosurgery usually rely simply on dose-volume or dose-diameter isoeffect curves. Due to the sparse clinical data available, these curves have typically not considered the target location in the brain, target histology, or treatment plan conformality as parameters in the calculation. In this study, a predictive model was generated to estimate the probability of cranial neuropathies as a result of acoustic schwannoma radiosurgery. METHODS AND MATERIALS: The dose-volume histogram reduction scheme was used to calculate the normal tissue complication probability (NTCP) from brainstem dose-volume histograms. The model's fitting parameters were optimized to provide the best fit to the observed complication data for acoustic neuroma patients treated with stereotactic radiosurgery at the University of Florida. The calculation was then applied to the remainder of the patients in the database. RESULTS: The best fit to our clinical data was obtained using n = 0.04, m = 0.15, and alpha/beta = 2.1 Gy(-1). Although the fitting parameter m is relatively consistent with ranges found in the literature, both the volume parameter, n, and alpha/beta are much smaller than the values quoted in the literature. The fit to our clinical data indicates that brainstem, or possibly a specific portion of the brainstem, is more radiosensitive than the parameters in the literature indicate, and that there is very little volume effect; in other words, irradiation of a small fraction of the brainstem yields NTCPs that are nearly as high as those calculated for entire volume irradiation. These new fitting parameters are specific to acoustic neuroma radiosurgery, and the small volume effect that we observe may be an artifact of the fixed relationship of acoustic tumors to specific regions of the brainstem. Applying the model to our patient database, we calculate an average NTCP of 7.2% for patients who had no cranial nerve complications, and the average NTCP for was 66% for patients who sustained a cranial neuropathy. For the entire patient population, the actual percentage of patients suffering either facial or trigeminal neuropathy was 14.7%, whereas the calculated average NTCP was 14.8%. DISCUSSION: NTCP calculations using brainstem dose-volume histograms can be used to estimate the rate of cranial neuropathies from acoustic neuroma radiosurgery. More clinical data and further study will lead to refinement of the model with time. PMID- 10837942 TI - Hypofractionated stereotactic radiotherapy as an alternative to radiosurgery for the treatment of patients with brain metastases. AB - PURPOSE: Modeling studies have demonstrated a potential biologic advantage of fractionated stereotactic radiotherapy for malignant brain tumors as compared to radiosurgery (SRS), even when only a few fractions are utilized. We prospectively evaluated the feasibility, toxicity, efficacy and cost of hypofractionated stereotactic radiotherapy (HSRT) in the treatment of selected radiosurgery eligible patients with brain metastases. METHODS AND MATERIALS: Patients with a limited number of brain metastases not involving the brainstem or optic chiasm underwent linac-based HSRT delivered in 3 fractions using a relocatable stereotactic frame. Depth-helmet and reference point measurements were recorded to address treatment accuracy. All patients underwent whole brain radiotherapy to a dose of 30 Gy. Toxicity, response, and survival duration were recorded for each patient. Prognostic factors were assessed by Cox regression analysis. Cost comparisons with a cohort of SRS treated patients were performed. RESULTS: Thirty two patients with 57 brain metastases were treated with HSRT. Twenty-three and 9 patients underwent HSRT for upfront and salvage treatment, respectively. The median dose delivered was 27 Gy, given in 3 fractions of 9 Gy. From 3328 depth helmet measurements, the absolute median setup deviation in AP, lateral, and vertical orientations was approximately 1.0 mm. No significant acute toxicity was seen. Late toxicities included seizures in four patients, and radionecrosis in two patients. The median survival duration from treatment was 12 months. KPS (p = 0.039) and RTOG-RPA class (p = 0.039) were identified as significant prognostic factors for survival. HSRT was $4119 less costly than SRS. CONCLUSION: HSRT, as delivered in this study, is more comfortable for patients and less costly than SRS in the treatment of selected patients with brain metastases. Proper dose selection and radiobiologic/toxicity trade-offs with SRS await further study. PMID- 10837943 TI - Four prognostic groups predict long-term survival from prostate cancer following radiotherapy alone on Radiation Therapy Oncology Group clinical trials. AB - PURPOSE: Gleason score (GS), T stage, and pathologic lymph node status have been described as major independent predictors of death due to prostate cancer in men treated with external beam radiotherapy (XRT). In this analysis we combine these three factors to define prognostic subgroups that correlate with disease-specific survival (DSS) death from prostate cancer. METHODS AND MATERIALS: Men entered on one of four Radiation Therapy Oncology Group (RTOG) Phase III randomized trials between 1975 and 1992, for clinically localized prostate cancer (CAP) (n = 1557), were selected for this analysis. Patients were included if: 1) they were evaluable, and eligible for the trial; 2) they received no hormonal therapy with their initial treatment; and 3) follow-up was available. For this study a DSS event was declared if: 1) death was certified as due to CAP; 2) death was due to complications of treatment; or 3) death was from unknown causes with active malignancy. The median follow-up for patients treated on early and late RTOG studies exceeded 11 and 6 years respectively. Subgroups were identified based on their pretreatment GS, T-stage, and lymph node such that patients with similar risk of dying from prostate cancer were combined. RESULTS: By combining patients with similar DSS, four subgroups were identified. Risk Group 1 patients had a GS = 2-6, and T1-2Nx; Group 2: GS = 2-6, T3Nx; or GS = 2-6, N+, or GS = 7, T1-2Nx; Group 3: T3Nx, GS = 7; or N+, GS = 7, or T1-2Nx, GS = 8-10; and Group 4 patients were T3Nx, GS = 8-10, or N+, GS = 8-10. The 5-, 10-, and 15-year DSS was 96%, 86%, and 72%; 94%, 75%, and 61%; 83%, 62%, and 39%; and 64%, 34%, and 27% for Groups 1 through 4, respectively. CONCLUSIONS: Recognition of these four risk groups provides a basis for estimating the long-term DSS for men treated with XRT alone and should facilitate the design of future prospective randomized trials. PMID- 10837944 TI - Predicting long-term survival, and the need for hormonal therapy: a meta-analysis of RTOG prostate cancer trials. AB - PURPOSE: To assess the impact of short-term and long-term androgen suppression on the disease-specific and overall survival of 2200 men treated with radiotherapy on one of 5 prospective randomized trials when stratified by prognostic risk groups. METHODS AND MATERIALS: Between 1975 and 1992, 2742 men were treated for clinically localized prostate cancer on one of 5 consecutive prospective Phase III randomized trials. Patients were selected for this analysis if they were deemed evaluable and eligible for the trial, and if follow-up information was available. For this analysis patients were stratified into four previously described prognostic risk groups: Group 1 patients had a Gleason score (GS) = 2 6, and T1-2Nx; Group 2: GS = 2-6, T3Nx; or GS = 2-6, N+, or GS = 7, T1-2Nx; Group 3: T3Nx, GS = 7; or N+, GS = 7, or T1-2Nx, GS = 8-10; and Group 4 patients were T3Nx, GS = 8-10, or N+, GS = 8-10. The median pretreatment prostate-specific antigen (PSA) was 25 ng/ml for the 434 evaluable patients for whom this information was available. The median follow-up times for patients treated on early studies exceeded 11 years, and for more recent studies 6 years. RESULTS: Risk group 2 patients with "bulky" or T3 disease appeared to have a disease specific survival benefit at 8 years with the addition of 4 months of goserelin and flutamide. Group 3 and 4 patients were noted to have an approximately 20% higher survival at 8 years with the addition of long-term hormonal therapy (p < or =0.0004). CONCLUSIONS: Based on this meta-analysis of RTOG trials, subsets of patients can be identified who either do not appear to benefit from the use of hormonal therapy, benefit from short-term hormonal therapy, or who benefit only from long-term hormonal therapy. These observations should be confirmed by prospective randomized trials before they can be considered conclusive. In the meantime, however, these observations provide rational guidelines for deciding who should receive hormonal therapy and for how long. PMID- 10837945 TI - Three-dimensional conformal therapy or standard irradiation in localized carcinoma of prostate: preliminary results of a nonrandomized comparison. AB - PURPOSE: We present preliminary results of a nonrandomized comparison of three dimensional conformal radiation therapy (3D CRT) and standard radiation therapy (SRT) in localized carcinoma of the prostate in two groups of patients with comparable prognostic factors treated during the same period. METHODS AND MATERIALS: Between January 1992 and December 1997, 146 patients were treated with 3D CRT and 131 with SRT alone for clinical stage T1c or T2 histologically confirmed carcinoma of the prostate. None of these patients received hormonal therapy. Mean follow-up for all patients is 3 years (range, 1-6 years). For 3D CRT, 7 intersecting fields were used (Cerrobend blocking or multileaf collimation) to deliver 68-73.8 Gy to the prostate; 3D dose distributions and dose-volume histograms (DVHs) of the planning target volume, bladder, and rectum were obtained. SRT consisted of bilateral 120 degrees rotational arcs, with portals with 2-cm margins around the prostate to deliver 68-70 Gy to the prostate. The criterion for chemical disease-free survival was a postirradiation prostate-specific antigen (PSA) (Tandem-R, Hybritech) value following the American Society for Therapeutic Radiology and Oncology guidelines. Symptoms during treatment were quantitated weekly, and late effects were assessed every 4 6 months. RESULTS: DVHs showed a two-thirds reduction in normal bladder or rectum receiving 70 Gy or more with 3D CRT. Higher 5-year chemical disease-free survival was observed with 3D CRT (91% for T1c and 96% for T2 tumors) compared with SRT (53% and 58%, respectively). There was no statistically significant difference in chemical disease-free survival in patients with Gleason score of 4 or less (p = 0.83), but with Gleason score of 5-7, the 5-year survival rates were 96% with 3D CRT and 53% with SRT (p < or = 0.01). In 111 patients with pretreatment PSA of 10 ng/mL or less, treated with 3D CRT, the chemical disease-free rate was 96% vs. 65% in 94 patients treated with SRT (p < or = 0.01). In patients with PSA of 10. 1-20 ng/mL, the chemical disease-free survival rate for 26 patients treated with 3D CRT was 88% compared with 40% for 20 patients treated with SRT (p = 0.05). The corresponding values were 71% and 26%, respectively, for patients with PSA levels of greater than 20 ng/mL (p = 0.30). On multivariate analysis, the most important prognostic factors for chemical failure were pretreatment PSA (p = 0. 023), nadir PSA (p = 0.001), and 3D CRT technique (p = 0.033). Moderate dysuria and difficulty in urinating were reported by 2-5% of patients treated with 3D CRT in contrast to 6-9% of patients treated with SRT; moderate urinary frequency and nocturia were reported by 18-24% treated with 3D CRT and 18-27% of patients in the SRT group. The incidence of moderate loose stools/diarrhea, usually after the 4th week of treatment, was 3-5% in the 3D CRT patients and 8-19% in the SRT group. Late intestinal morbidity (proctitis, rectal bleeding) was very low (1.7%) in the 3D CRT group in contrast to the SRT patients (8%). CONCLUSION: Three dimensional CRT spares more normal tissues, yields higher chemical disease-free survival, and results in less treatment morbidity than SRT in treatment of Stage T1-T2 prostate cancer. Longer follow-up is needed to confirm these preliminary observations. PMID- 10837946 TI - Radiotherapy of prostate cancer with or without intensity modulated beams: a planning comparison. AB - PURPOSE: To evaluate whether intensity modulated radiotherapy (IMRT) by static segmented beams allows the dose to the main portion of the prostate target to escalate while keeping the maximal dose at the anterior rectal wall at 72 Gy. The value of such IMRT plans was analyzed by comparison with non-IMRT plans using the same beam incidences. METHODS AND MATERIALS: We performed a planning study on the CT data of 32 consecutive patients with localized adenocarcinoma of the prostate. Three fields in the transverse plane with gantry angles of 0 degrees, 116 degrees, and 244 degrees were isocentered at the center of gravity of the target volume (prostate and seminal vesicles). The geometry of the beams was determined by beam's eye view autocontouring of the target volume with a margin of 1.5 cm. In study 1, the beam weights were determined by a human planner (3D-man) or by computer optimization using a biological objective function with (3D-optim-lim) or without (3D-optim-unlim) a physical term to limit target dose inhomogeneity. In study 2, the 3 beam incidences mentioned above were used and in-field uniform segments were added to allow IMRT. Plans with (IMRT-lim) or without (IMRT-unlim) constraints on target dose inhomogeneity were compared. In the IMRT-lim plan, target dose inhomogeneity was constrained between 15% and 20%. After optimization, plans in both studies were normalized to a maximal rectal dose of 72 Gy. Biological (tumor control probability [TCP], normal tissue complication probability [NTCP]) and physical indices for tumor control and normal tissue complication probabilities were computed, as well as the probability of the uncomplicated local control (P+). RESULTS: The IMRT-lim plan was superior to all other plans concerning TCP (p < 0.0001). The IMRT-unlim plan had the worst TCP. Within the 3D plans, the 3D-optim-unlim had the best TCP, which was significantly different from the 3D-optim-lim plan (p = 0.0003). For rectal NTCP, both IMRT plans were superior to all other plans (p < 0.0001). The IMRT-unlim plan was significantly better than the IMRT-lim plan (p < 0.0001). Again, 3D-optim-unlim was superior to the other 3D plans (p < 0. 0007). Physical endpoints for target showed the mean minimal target dose to be the lowest in the IMRT-unlim plan, caused by a large target dose inhomogeneity (TDI). Medial target dose, 90th percentile, and maximal target dose were significantly higher in both IMRT plans. Physical endpoints for the rectum showed the IMRT-unlim plan to be superior compared to all other plans. There was a strong correlation between the 65th percentile (Rp65) and rectal NTCP (correlation coefficient > or =89%). For bladder, maximal bladder dose was significantly higher in the IMRT-unlim plan compared to all other plans (p < or = 0.0001).P+ was significantly higher in both IMRT-plans than in all other plans. The 3D-optim-unlim plan was significantly better than the two other 3D plans (p < 0.0001). CONCLUSION: IMRT significantly increases the ratio of TCP over NTCP of the rectum in the treatment of prostate cancer. However, constraints for TDI are needed, because a high degree of TDI reduced minimal target dose. IMRT improved uncomplicated local control probability. In our department, IMRT by static segmented beams is planned and delivered in a cost-effective way. IMRT-lim has replaced non-modulated conformal radiotherapy as the standard treatment for prostate cancer. PMID- 10837947 TI - Defining the appropriate radiation dose for pretreatment PSA < or = 10 ng/mL prostate cancer. AB - PURPOSE: To investigate whether a dose response exists for biochemical no evidence of disease (bNED) control in prostate cancer patients with pretreatment prostate-specific antigen (PSA) < or = 10 ng/mL and to identify the patient subgroups affected. METHODS AND MATERIALS: Between 5/89 and 10/97, 488 T1-T3 NX-0 M0 prostate cancer patients with PSA < or = 10 ng/mL were treated with three dimensional conformal radiation therapy (3D-CRT) alone. Median and mean pretreatment PSA values were 6.3 and 6.2, respectively. Gleason scores of 2-6 and 7-10 were noted in 386 and 102 men, respectively. AJCC 1992 palpation T1-T2AB tumors were noted in 415 patients. Perineural invasion (PNI) was noted in 60 men. Mean and median age was 67 and 68 years, respectively. Dose to the center of the prostate ranged from 6260 cGy to 8409 cGy with a mean and median of 7423 cGy and 7278 cGy, respectively. Patients were stratified into three groups according to dose: <7250 cGy, 7250-7599 cGy, and > or =7600 cGy. Median dose in these three groups was 7067 cGy, 7278 cGy, and 7734 cGy, respectively. Univariate analysis was performed to determine differences in bNED control (American Society for Therapeutic Radiology and Oncology [ASTRO] Consensus Guidelines definition of failure) by dose group for the entire cohort, for 310 good prognosis patients (T1 T2A, Gleason score 2-6, absence of PNI), and for 178 poor prognosis patients (T2B T3 or Gleason score 7-10 or presence of PNI) (1). Multivariate analysis (MVA) was performed to determine if dose was an independent predictor of bNED control. Median follow-up was 36 months. RESULTS: A dose response was not demonstrated for the entire group of patients with pretreatment PSA < or =10 ng/mL. Doses of <7250 cGy, 7250-7599 cGy, and > or =7600 cGy were associated with 5-year bNED control rates of 73%, 86%, and 89%, respectively (p = 0.12). MVA demonstrated prognosis group (p = 0. 038) to be the only independent predictor of bNED control. Good prognosis patients had a 5-year bNED of 85% and no dose response was seen. The subgroup of poor prognosis patients demonstrated a 5-year bNED control rate of 81% and a dose response was seen for those receiving > or =7600 cGy, compared to the two lower dose groups (94% vs. 75% vs. 70%; p = 0.0062). MVA for the poor prognosis subset demonstrated dose (p = 0.01) to be the only independent predictor for improved bNED control. CONCLUSIONS: The poor prognosis subset of PSA < or =10 ng/mL prostate cancer patients benefit from dose escalation. A dose response is not demonstrated for prostate cancer patients with pretreatment PSA < or =10 ng/mL and other favorable features. PMID- 10837948 TI - Prostate position late in the course of external beam therapy: patterns and predictors. AB - PURPOSE: To examine prostate and seminal vesicles position late in the course of radiation therapy and to determine the effect and predictive value of the bladder and rectum on prostate and seminal vesicles positioning. METHODS AND MATERIALS: Twenty-four patients with localized prostate cancer underwent a computerized tomography scan (CT1) before the start of radiation therapy. After 4-5 weeks of radiation therapy, a second CT scan (CT2) was obtained. All patients were scanned in the supine treatment position with instructions to maintain a full bladder. The prostate, seminal vesicles, bladder, and rectum were contoured. CT2 was aligned via fixed bony anatomy to CT1. The geometrical center and volume of each structure were obtained and directly compared. RESULTS: The prostate shifted along a diagonal axis extending from an anterior-superior position to a posterior inferior position. The dominant shift was to a more posterior-inferior position. On average, bladder and rectal volumes decreased to 51% (+/-29%) and 82% (+/-45%) of their pretreatment values, respectively. Multiple regression analysis (MRA) revealed that bladder movement and volume change and upper rectum movement were independently associated with prostate motion (p = 0.016, p = 0. 003, and p = 0.052 respectively). CONCLUSION: Patients are often instructed to maintain a full bladder during a course of external beam radiation therapy, in the hopes of decreasing bladder and small bowel toxicity. However, our study shows that large bladder volumes late in therapy are strongly associated with posterior prostate displacement. This prostate displacement may result in marginal miss. PMID- 10837949 TI - The impact of postimplant edema on the urethral dose in prostate brachytherapy. AB - PURPOSE: The objective of this work is to determine the effect of timing of the postimplant CT scan on the assessment of the urethral dose. METHODS AND MATERIALS: A preimplant CT scan and two postimplant CT scans were obtained on 50 patients who received I-125 prostate seed implants. The first postimplant CT scan was obtained on the day of the implant; the second usually 4 to 9 weeks later (mean: 46 +/- 23 days; range: 27-135 days). The urethra was localized in each postimplant CT scan and a dose-volume histogram (DVH) of the urethral dose was compiled from each CT study. The relative decrease in the prostate volume between the first and second postimplant CT scans was determined by contouring the prostate in each CT scan. RESULTS: The prostate volume decreased by 27 +/- 9% (mean +/- SD) between the first and second postimplant CT scans. As a result, the averaged urethral dose derived from the second CT scan was about 30% higher. In terms of dose, the D(10), D(25), D(50), D(75), and D(90) urethral doses derived from the second CT scan were 90 +/- 56 Gy, 81 +/- 49 Gy, 67 +/- 42 Gy, 49 +/- 44 Gy, and 40 +/- 46 Gy higher, respectively. The increase in the urethral dose is correlated with the decrease in the prostate volume (R = 0.57, rho < 0.01). CONCLUSION: The assessment of the urethral dose depends upon the timing of the postimplant CT scan. The mean D(10) dose derived from the CT scans obtained at 46 +/- 23 days postimplant was 90 +/- 56 Gy higher than that derived from the CT scans obtained on the day of the implant. Because of this large difference, the timing of the postimplant CT scan needs to be specified when specifying dose thresholds for urethral morbidity. PMID- 10837950 TI - High-dose-rate brachytherapy as sole modality for early-stage endobronchial carcinoma. AB - PURPOSE: To evaluate exclusive high-dose-rate brachytherapy for localized early stage non-small-cell bronchial carcinoma; to develop new insights in treatment catheter positioning and tumor-volume assessment by computed tomography (CT) scan. METHODS AND MATERIALS: Between 1992 and 1996, 34 patients with non-small cell bronchial carcinoma were treated by brachytherapy alone. All patients were medically inoperable and had contraindications for external beam irradiation. The treatment protocol was six sessions of 5 Gy over 6 weeks. The treatment catheter was placed under fiberoscopy and was positioned with the help of spacer catheters or with a surrounding plastic tube; CT scan was performed in 50% of the cases to measure the spacing between the applicator and the bronchial wall. Dose prescription was individually based on clinical and radiologic evaluation of tumor volume. RESULTS: Local disease failure occurred in 5 patients (15%). With a median follow-up of 2 years, the local control rate was 85% and the survival rate 78%. No acute toxicity was found, except one pneumothorax. CONCLUSION: Brachytherapy alone can give an optimal therapeutic ratio in small endobronchial carcinomas without radiation-induced morbidity. Such results are achieved after careful tumor volume evaluation and individualized treatment catheter positioning. PMID- 10837951 TI - Treatment of roentgenographically occult endobronchial carcinoma with external beam radiotherapy and intraluminal low-dose-rate brachytherapy: second report. AB - PURPOSE: To evaluate the efficacy and toxicity of treatment with external beam radiotherapy and intraluminal low-dose-rate brachytherapy for roentgenographically occult endobronchial carcinoma (ROEC). METHOD AND MATERIALS: A total of 79 lesions (71 cases) of ROEC were treated with external beam radiotherapy and intraluminal low-dose-rate brachytherapy from July 1991 to December 1998. Of these lesions, 68 (64 cases) were treated with our standard dose (external beam radiotherapy of 40 Gy and intraluminal low-dose-rate brachytherapy of 25 Gy) and are the subject of this report. All 64 patients were males, and their ages ranged from 55 to 80 (median, 68) years. The histologic diagnosis was squamous cell carcinoma in all patients. RESULTS: In all cases, the scheduled treatment was carried out within 2 months. Follow-up period ranged from 4 to 91 (median, 44) months from the beginning of this treatment. Acute toxicity was tolerable. Grade 2 radiation pneumonitis was observed in 4 cases, and there was no case of greater than Grade 2 radiation fibrosis. Nineteen cases of bronchial stenosis and 23 cases of bronchial obstruction were observed on follow up bronchoscopy. However, no Grade 2 or greater deterioration of respiratory function due to radiotherapy, prolonged symptoms, or fatal toxicity was observed. Nine patients suffered recurrence, 5 of whom were rescued by surgery and external beam radiotherapy, and 4 of whom died of disease. The 5-year cause-specific survival, overall-survival, and disease-free rate were 96.1%, 72.3%, and 87.3%, respectively. CONCLUSION: Combined treatment with external beam radiotherapy and intraluminal low-dose-rate brachytherapy is effective and results in acceptable complications for ROEC. PMID- 10837952 TI - Radiation dose-effect relations and local recovery in perfusion for patients with non-small-cell lung cancer. AB - PURPOSE: To determine local dose-effect relations for lung perfusion and density changes due to irradiation for patients with non-small-cell lung cancer (NSCLC) and to quantify the effect of reperfusion. METHODS AND MATERIALS: For 25 NSCLC patients and a reference group of 81 patients with healthy lungs, registered single photon emission computed tomography (SPECT) lung perfusion and CT scans were made, before and after radiotherapy. Average dose-effect relations for perfusion and CT-density changes were calculated and compared with the dose effect relation of the reference group. On the basis of these dose-effect relations, the post-RT perfusion was predicted for each patient and compared to the measured post-RT perfusion. RESULTS: Well-perfused lung regions of the NSCLC patients showed the same dose-effect relation as the reference patients. By comparing predicted and measured post-treatment perfusion scans, regions of reperfusion could be determined for 18 of 25 NSCLC patients but for none of the reference patients. CONCLUSION: Well-perfused lung tissue of patients with NSCLC behaves like healthy lung tissue with respect to radiation. The dose-effect relation for perfusion and CT density was extended for doses up to 80 Gy. Radiation damage in poorly perfused lung regions was less than predicted as a consequence of local reperfusion. PMID- 10837953 TI - Response of global quality of life to high-dose palliative radiotherapy for non small-cell lung cancer. AB - PURPOSE: To estimate the impact of high-dose palliative radiotherapy treatment (RT) for inoperable non-small cell lung cancer (NSCLC) on the patient's quality of life (QoL) over the remaining survival period, and to compute the number of quality adjusted life days (QALDs) gained. METHODS AND MATERIALS: The QoL of an NSCLC patient is modelled as a function of the days left to live, the days since the start of treatment, a patient specific intercept, and a random error term. Least squares regression analysis is used to fit this model to 376 monthly QoL observations supplied by 42 prospectively enrolled high-dose palliative RT patients with NSCLC. Prediction analysis, based on the regression results and on previously published estimates of the survival response to high-dose palliative RT, is used to compute QALDs gained as a result of treatment. RESULTS: QoL improves steadily over the first 86 days after the start of treatment. This improvement then dissipates to 0 over the subsequent 140 days. Median survival after entry (266 days) yields 158.5 QALDs, of which 56.9 can be attributed treatment: 12.6 as a result of a higher daily QoL and 44.3 as a result of longer survival. CONCLUSION: Patients with inoperable NSCLC who received high-dose palliative RT, and survived the median 266 days after entry into the study, can attribute about one-third of their 158.5 QALDs to the QoL and survival responses to treatment. PMID- 10837954 TI - Radiation treatment for cervical esophagus: patterns of practice study in Canada, 1996. AB - PURPOSE: To assess the patterns of practice among Canadian radiation oncologists who treat esophageal cancers, using a trans-Canada survey, completed at the end of 1996. METHODS AND MATERIALS: One of 3 case presentations of different stages of cervical esophageal cancer was randomly assigned and sent to participating radiation oncologists by mail. Respondents were asked to fill in questionnaires regarding treatment techniques and to outline target volumes for the boost phase of radiotherapy. Radiation oncologists from 26 of 27 (96%) of all Canadian centers participated. RESULTS: High-energy X-rays (>/= 10 MV) were employed by 68% of the respondents in part of the treatment course. The majority (83%) of the radiation oncologists used at least two phases of treatment. Very few, 10 of 59 (17%), responses started with multifield treatment. The most frequently used prescription dose was 60 Gy/30 fractions/6 weeks, given with concurrent chemotherapy. Dose prescriptions were to the isocenter in 39 of 48 (81%) or to a particular isodose line in 9 of 48 (19%) of respondents. CONCLUSION: There was a variety of radiation treatment techniques in this trans-Canada survey. The majority of the patients had combined cisplatin-based chemoradiation. The isocenter was not used consistently as a dose prescription point. PMID- 10837956 TI - A pilot study of preoperative continuous infusion 5-fluorouracil, external microwave hyperthermia, and external beam radiotherapy for treatment of locally advanced, unresectable, or recurrent rectal cancer. AB - PURPOSE: To determine the feasibility of combining external beam radiotherapy, continuous infusion 5-fluorouracil (5-FU), and external microwave hyperthermia in patients with locally advanced, unresectable, or recurrent adenocarcinoma of the rectum. METHODS AND MATERIALS: From 7/95 through 2/99, 15 patients were enrolled in the study. The treatment regimen consisted of continuous infusion 5-FU 250 mg/m(2)/d 7 days/week beginning on day 1, external beam radiotherapy to the pelvis, 4500 cGy, 180 cGy/d 5 days/week using a 3 or 4-field technique, and external microwave hyperthermia on days 3, 8, 15, 22, and 29. Chemotherapy was stopped on the last day of radiotherapy. Surgical resection, if feasible, was scheduled 3-6 weeks after completing thermochemoradiotherapy. For this regimen to be considered feasible, no more than 2 of the 15 patients should fail to complete therapy due to life-threatening toxicity. Toxicity was scored using National Cancer Institute Criteria. RESULTS: All patients completed the chemoradiotherapy portion of the protocol. Eleven of the 15 patients completed all 5 hyperthermia treatments. Of the 4 patients who did not receive the full course of hyperthermia, only 1 patient had treatment stopped due to life-threatening toxicity. The other 3 patients did not complete hyperthermia due to scheduling errors (n = 2) or patient request (n = 1). Five of 15 patients required a treatment interruption due to toxicity > or = Grade 3. Seven patients experienced lesser degrees of toxicity which did not require treatment interruption. Three patients experienced no side effects. The most common toxicities were dermatitis and diarrhea. Of the 14 patients in whom surgery was planned, 11 (79%) were resectable. There was one pathologic complete response. CONCLUSIONS: It is feasible to deliver thermochemoradiotherapy, as prescribed in this study, to patients with locally advanced, unresectable, or recurrent rectal cancer. The therapy is moderately toxic, with one-third of patients requiring temporary treatment interruptions. The regimen appears active against rectal cancer, and appears to warrant further consideration as a treatment option for this patient population. PMID- 10837955 TI - Prospective trial of preoperative concomitant boost radiotherapy with continuous infusion 5-fluorouracil for locally advanced rectal cancer. AB - RATIONALE: To evaluate the response to a concomitant boost given during standard chemoradiation for locally advanced rectal cancer. METHODS AND MATERIALS: Concomitant boost radiotherapy was administered preoperatively to 45 patients with locally advanced rectal cancer in a prospective trial. Treatment consisted of 45 Gy to the pelvis with 18 mV photons at 1.8 Gy/fraction using a 3-field belly board technique with continuous infusion 5FU chemotherapy (300mg/m(2)) 5 days per week. The boost was given during the last week of therapy with a 6-hour inter-fraction interval to the tumor plus a 2-3 cm margin. The boost dose equaled 7.5 Gy/5 fractions (1.5 Gy/fraction); a total dose of 52.5 Gy/5 weeks was given to the primary tumor. Pretreatment tumor stage, determined by endorectal ultrasound and CT scan, included 29 with T3N0 [64%], 11 T3N1, 1 T3Nx, 2 T4N0, 1 T4N3, and 1 with TxN1 disease. Mean distance from the anal verge was 5 cm (range 0-13 cm). Median age was 55 years (range 33-77 years). The population consisted of 34 males and 11 females. Median time of follow-up is 8 months (range 1-24 months). RESULTS: Sphincter preservation (SP) has been accomplished in 33 of 42 (79%) patients resected to date. Three patients did not undergo resection because of the development of metastatic disease in the interim between the completion of chemoradiation (CTX/XRT) and preoperative evaluation. The surgical procedures included proctectomy and coloanal anastomosis (n = 16), low anterior resection (n = 13), transanal resection (n = 4). Tumor down-staging was pathologically confirmed in 36 of the 42 (86%) resected patients, and 13 (31%) achieved a pathologic CR. Among the 28 tumors (67%) located <6 cm from the anal verge, SP was accomplished in 21 cases (75%). Although perioperative morbidity was higher, toxicity rates during CTX/XRT were comparable to that seen with conventional fractionation. Compared to our contemporary experience with conventional CTX/XRT (45Gy; 1.8 Gy per fraction), improvements were seen in SP (79% vs. 59%; p = 0.02), SP for tumors <6 cm from the anal verge (75% vs. 42%; p = 0.003), and down staging (86% vs. 62%; p = 0.003). CONCLUSION: The SP rate with concomitant boost radiation has been highly favorable with rates of response which are higher than those previously reported for chemoradiation without administration of a boost. Further evaluation of this radiotherapeutic strategy appears warranted. PMID- 10837957 TI - Phase II trial of adjuvant radiation and intraperitoneal 5-fluorouracil for locally advanced colon cancer: results with 10-year follow-up. AB - PURPOSE: To determine the toxicity, disease-free survival, and overall survival for patients with Modified Astler-Coller (MAC) B2-3 or C1-3 colon cancer receiving adjuvant radiation and sequential intraperitoneal 5-fluorouracil (5 FU). METHODS AND MATERIALS: From August 1984 to June 1989, 45 patients were accrued to this Phase II trial and received a 21-week course of intraperitoneal 5 FU (20 mg/kg/d x 5) and external beam radiation. The radiation was delivered to the tumor bed and para-aortic lymph nodes in two split-courses of 22.5 Gy, alternating with the first two cycles of chemotherapy. All patients then received 4 additional cycles of intraperitoneal 5-FU. RESULTS: The therapy was well tolerated with 4 patients experiencing Grade 3 peritonitis. Four patients developed small bowel obstruction requiring surgery; in each instance, recurrent tumor was found at the time of laparotomy. The median and overall survivals at 10 years were 9.3 months and 53% respectively. Local failures were infrequent, occurring in only 11% of patients treated. CONCLUSIONS: Sequential intraperitoneal 5-FU and tumor-bed/para-aortic irradiation is tolerable in patients with resected colon cancer. Although the incidence of local and regional relapse appeared to be lower than anticipated, this did not appear to translate into improved survival. PMID- 10837958 TI - Salvage extended-field irradiation in follicular non-Hodgkin's lymphoma after failure of chemotherapy. AB - PURPOSE: To evaluate the efficacy of total abdominopelvic (TAI) and total body irradiation (TBI) in heavily pretreated follicular non-Hodgkin's lymphoma (NHL). PATIENTS AND METHODS: From 1983 to 1998, 34 patients received TAI (n = 22) or TBI (n = 12). All had Stage III or IV, Class B, C, D NHL in the working formulation and failed after receiving 1-5 regimens of chemotherapy. TAI was given at 20 Gy over a 3-week period. TBI was delivered in two successive half-body irradiations of 15 Gy over a 2-week period with a 4-week interval between each. RESULTS: Mean follow-up from TAI or TBI was 120 months (range, 6-180). Seventy-six percent of patients achieved complete response and 24% partial response. Median survival was 62 months, 5-year and 10-year overall survival was 59% and 41%, and disease-free survival was 56% and 30%, respectively. Grade III or IV toxicity was gastrointestinal in 38% of patients and hematologic in 30%. No toxic death or delayed complications were observed. CONCLUSION: Extended-field irradiation is feasible and efficient after failure of chemotherapy in follicular NHL. PMID- 10837959 TI - An alternative mantle irradiation technique using 3D CT- based treatment planning for female patients with Hodgkin's disease. AB - PURPOSE: For female patients, radiotherapy treatment for Hodgkin's disease invariably results in the irradiation of breast tissue that may lead to radiation induced secondary cancers. The risk for secondary breast cancer is correlated with dose. We have developed a technique in an attempt to increase breast sparing during mantle field irradiation for female patients. MATERIAL AND METHODS: To minimize the irradiated breast volume, a virtual simulation technique making use of a Styrofoam breast immobilization board has been developed whereby the patient lies prone with the breasts positioned in grooves within the board. The breast position is adjusted using Styrofoam wedges, and breast placement is verified using an AP CT-pilot view. A CT scan of the neck and thoracic regions is taken, and the lymph nodes, breast volume and critical structures are outlined. Virtual simulation of the mantle fields (typically AP/PA isocentric beams) is performed, and beam blocks are drawn on the digitally reconstructed radiographs (DRR) generated by the virtual simulation package. The shielding is designed to allow adequate margins around the lymph nodes while maximizing shielding of the lung and breast tissues. The para-aortic fields are also easily determined through virtual simulation, where multi-planar reconstructions (MPR) and 3D renderings of the patient's CT data are used to determine the field limits and beam gaps. In addition to allowing for the geometric optimization of the positioning of the breasts under the lung shields, the virtual simulation technique provides the necessary information for a 3D dosimetric analysis, including dose-volume histograms (DVHs) of the irradiated breast volume. RESULTS: The 3D breast sparing technique was qualitatively and quantitatively compared to non-CT-based techniques and other 3D techniques currently available to assess the protection of the breasts. In a preliminary analysis, virtual simulation images (DRRs, 3D rendering and multi-planar reconstruction) demonstrated the advantage of using the breast sparing technique. A further analysis of DVHs showed a reduction of at least 50% in the volume of breast tissue irradiated when using the breast positioning board and virtual simulation as compared to the conventional simulation techniques where a breast immobilization board was not used. CONCLUSIONS: The use of a breast immobilization board and of a virtual simulation technique is recommended for the planning and treatment of female patients with Hodgkin's disease. DVH analysis has shown that this leads to a decrease in the volume of breast irradiated. It is hoped that this approach will reduce the risk of secondary breast malignancies in female patients with Hodgkin's disease. PMID- 10837960 TI - A phase I/II study of hypofractionated whole abdominal radiation therapy in patients with chemoresistant ovarian carcinoma: Karnofsky score determines treatment outcome. AB - PURPOSE: Radiation therapy can provide useful palliation in chemorefractory ovarian cancer patients. The purpose of this study was to prospectively study the palliative effect of a hypofractionated radiation treatment regimen. Change in quality-of-life scores (Functional Assessment of Cancer Therapy [FACT], Karnofsky scale), pain score, and tolerance to therapy were also assessed. METHODS AND MATERIALS: A single-institution Phase I/II trial was initiated in patients with chemoresistant recurrent or progressive ovarian cancer. All patients had symptomatic and measurable intra-abdominal disease. Patients were treated with a single radiation fraction (700 cGy) or two fractions (300 cGy twice a day) to the whole abdomen over 1 day. Quality-of-life scale (FACT G version 2) was assessed at baseline and 1 and 3 months following treatment. Karnofsky scale and pain score were also evaluated in the same time frame. RESULTS: Sixteen patients were prospectively entered into this protocol between February 1996 and September 1998. Twelve patients received a single 700 cGy fraction and four 300 cGy twice a day. All were heavily pretreated and 9 (56%) had a poor performance status prior to treatment. Symptoms needing palliation included pain (14), ascites (10), and bleeding (2). Symptomatic improvement occurred in all patients with pain (5 complete response [CR] and 7 partial response [PR], all patients with bleeding (CR 2), and two (20%) with ascites. Five patients (31%) had a reduction in lesion size documented radiologically in three. The mean duration of response was 22 weeks in patients with a Karnofsky score >70. Thirteen patients developed transient nausea and vomiting which resolved in 48 hours in all. All patients developed a transient lymphopenia. Thirteen patients completed a follow-up quality-of-life scale. There was an improvement in the physical and functional components of the scale in patients with Karnofsky score of 90-100. There was no improvement in quality of life in patients with Karnofsky score <90 despite symptomatic response. Median survival was 3 months from the date of treatment. CONCLUSION: Hypofractionated radiation therapy is an effective palliative treatment for end-stage ovarian cancer patients. Karnofsky score can help determine the duration of response and survival for this high-risk population. PMID- 10837961 TI - Variability of the depth of supraclavicular and axillary lymph nodes in patients with breast cancer: is a posterior axillary boost field necessary? AB - PURPOSE: To determine the variability of the depth of supraclavicular (SC) and axillary (AX) lymph nodes in patients undergoing radiation therapy for breast cancer and to relate this variability with the patient's anterior/posterior (A/P) diameter. The dosimetric consequences of the variability in depth are explored and related to the need for a posterior axillary boost field. METHOD AND MATERIALS: In 49 patients undergoing treatment-planning computed tomography (CT) scanning in the treatment position, the maximum depth of the SC and AX lymph nodes was measured on CT images. The A/P diameter was measured at the location of the SC and AX, respectively. The relationship between the SC/AX lymph node depth and patient diameter was determined using linear regression. For an anterior SC and AX field, the relative dose to the SC and AX lymph nodes were calculated for a 6 MV photon beam. RESULTS: The maximum depth of the SC lymph nodes ranged from 2.4 to 9.5 cm (median, 4.3 cm). The depth was less than 3 cm in 4 patients, 3-6 cm in 39 (80%), and greater than 6 cm in 6 patients. There was a linear relationship between the SC lymph node depth and the A/P diameter. The depth of the SC lymph nodes in cm equals approximately one-half of the A/P diameter minus 3.5 (r(2) = 0.69). In 94% (46 of 49) of patients, the SC lymph node depth was between one-fifth and one-half of the A/P diameter. The depth of the axillary lymph nodes ranged from 1.4 to 8 cm (median, 4.3 cm). The depth was less than 3 cm in 8 patients, 3-6 cm in 32 (65%), and greater than 6 cm in 9 patients. The AX lymph node depth in cm equals approximately one-half of the A/P diameter minus 3 (r(2) = 0.81). In all patients, the AX lymph nodes were shallower than mid-depth. The depth of the SC and AX lymph nodes was within +/- 1 cm in 53% (26 of 49) of patients. The AX lymph nodes were located at >/= 1 cm shallower or greater depth than the SC in 24.5% (12 of 49) and 22.5% (11 of 49) of patients, respectively. If an anterior 6-MV beam only is used to treat the SC and AX lymph nodes in these 49 patients, the dose to the AX is within +/- 5% of the SC dose in 53% (26 of 49) patients and is 90% or more of the dose delivered in the SC in 90% (44 of 49) of patients. CONCLUSION: The maximum depth of the SC and AX lymph nodes varies widely and is related to the patient's size represented by the A/P diameter. In most patients, the AX lymph nodes lie at approximately the same depth or shallower than the SC. Therefore, the rationale for a posterior axillary boost field needs to be further assessed. When the AX and SC lymph nodes are deep, opposed supraclavicular and axillary fields and/or the use of a higher energy beam might be reasonable. PMID- 10837962 TI - Open magnetic resonance imaging using titanium-zirconium needles: improved accuracy for interstitial brachytherapy implants? AB - PURPOSE: To evaluate the benefit of using an open magnetic resonance (MR) machine and new MR-compatible needles to improve the accuracy of brachytherapy implants in pelvic tumors. METHODS AND MATERIALS: The open MR machine, foreseen for interventional procedures, allows direct visualization of the pelvic structures that are to be implanted. For that purpose, we have developed MR- and CT compatible titanium-zirconium (Ti-Zr) brachytherapy needles that allow implantations to be carried out under the magnetic field. In order to test the technical feasibility of this new approach, stainless steel (SS) and Ti-Zr needles were first compared in a tissue-equivalent phantom. In a second step, two patients implanted with Ti-Zr needles in the brachytherapy operating room were scanned in the open MR machine. In a third phase, four patients were implanted directly under open MR control. RESULTS: The artifacts induced by both materials were significantly different, strongly favoring the Ti-Zr needles. The implantation in both first patients confirmed the excellent quality of the pictures obtained with the needles in vivo and showed suboptimal implant geometry in both patients. In the next 4 patients, the tumor could be punctured with excellent accuracy, and the adjacent structures could be easily avoided. CONCLUSION: We conclude that open MR using MR-compatible needles is a very promising tool in brachytherapy, especially for pelvic tumors. PMID- 10837963 TI - Different mechanisms between premitotic apoptosis and postmitotic apoptosis in X irradiated U937 cells. AB - PURPOSE: Apoptosis is currently being evaluated for its importance as a pathway of radiation-induced cell death. However, the difference in the mechanisms between premitotic and postmitotic apoptosis following X-irradiation remains not well understood. We show here that the human monoblastoid cell line U937 can be induced to undergo these two different types of apoptosis. METHODS AND MATERIALS: U937 cells were irradiated at a dose of 5 or 20 Gy, and the DNA fragmentation rate was measured by both flow cytometric analysis and gel electrophoresis. Activation of caspase-3 was detected by Western blot analysis and fluorogenic assay using acetyl-Asp-Glu-Val-Asp-7-amino-4-methyl-coumarin (Ac-DEVD-AMC). Detection of mitochondrial transmembrane potential (DeltaPsi) was performed by using Rho123. Chasing of S-phase fraction following X-irradiation was performed after labeling with 5-bromo-2'-deoxyuridine (BrdU). Thymidine was used for synchronization of the cells. Inhibition of caspase-3 activity was achieved by Acetyl-Asp-Glu-Val-Asp-aldehyde (Ac-DEVD-CHO). RESULTS: Time courses of the apoptotic rates, caspase activation, and DeltaPsi indicated that two different types of cell death were induced by the different X-ray doses. High-dose X-ray (20 Gy) induced a rapid and strong apoptosis, whereas low-dose X-ray (5 Gy) induced a slow and mild apoptosis. Cell-cycle analyses revealed that there was cell death before cell division in the former apoptosis but the cells must be dying after cell division in the latter apoptosis. By means of cell-cycle synchronization, the S-phase cells proved to be the most sensitive fraction to premitotic apoptosis, but an obvious difference in the susceptibility to cell death among the cell-cycle phases was not observed in postmitotic apoptosis. Ac DEVD-CHO treatment effectively blocked caspase activity and premitotic apoptosis, but it failed to block postmitotic apoptosis. CONCLUSIONS: Irradiation of U937 cells at different X-ray doses induced two different types of apoptotic cell death, premitotic apoptosis and postmitotic apoptosis, which are characterized by the time course and cell-cycle specificity. Decision concerning these two types of apoptotic cell death may be made by the difference in the magnitude of cell damage following X-irradiation. PMID- 10837964 TI - Survival of human glioma cells treated with various combination of temozolomide and X-rays. AB - PURPOSE: To investigate the effect of temozolomide, a 3-methyl derivative of mitozolomide in combination with X-rays in human glioma-derived cell lines. METHODS AND MATERIALS: Glioma cell lines D384 and U251 were treated with temozolomide for various periods of time in combinations with X-rays. Temozolomide administration was repeated every 24 h for exposures up to 96 h. Cytotoxicity was determined with a clonogenic assay. RESULTS: Incubation of D384 cells with temozolomide during 24 h prior to or following irradiation results in a moderate enhancement of the cytotoxicity. Prolonged treatment with temozolomide, i.e., 48-96 h before X-rays, causes a stronger potentiation. In contrast, no enhancement is observed in irradiated U251 cells in combination with 24-96 h temozolomide treatment. In addition to single-dose irradiation, we investigated the effect of temozolomide in D384 cells with concomitant fractionated irradiation. A 96-h exposure to temozolomide with simultaneous doses of 2 Gy X-rays at 24-h intervals, causes a significant further reduction in cell survival as compared to fractionated irradiation only. CONCLUSION: Depending on the cell line, treatment of glioma cells with temozolomide and X-rays can have either an additional effect or potentiate cell killing. PMID- 10837965 TI - Radiosensitization produced in vivo by once- vs. twice-weekly 2'2'-difluoro-2' deoxycytidine (gemcitabine). AB - PURPOSE: Gemcitabine (2'2'-difluoro-2'-deoxycytidine, dFdCyd) is a potent radiosensitizer of rodent and human tumor cells. Our Phase I clinical trial using once-weekly dFdCyd as a radiosensitizer in the treatment of patients with Stage IV squamous cell head and neck cancer has produced a high rate of tumor response and significant normal mucosal toxicity. These findings raised the question of whether we are using dFdCyd in the optimal dose and schedule. In vitro studies suggest that twice-weekly dFdCyd has the potential to be more effective than once weekly dFdCyd when administered in combination with radiation (RT) given 5 days per week. Therefore, we have used a mouse model to assess whether the therapeutic ratio of combined modality therapy may be improved by using a twice-weekly drug regimen. We asked two questions: 1) Does a once-weekly or twice-weekly dFdCyd regimen cause more normal tissue radiosensitization? 2) Does a once-weekly or twice-weekly dFdCyd + RT regimen produce a better therapeutic index? METHODS AND MATERIALS: To assess normal tissue toxicity, C3H mice underwent mouth (60)Co RT (27.5 Gy in 5 daily fractions) +/- dFdCyd delivered intraperitoneally (IP) either once or twice weekly 6 hours prior to irradiation. Acute lip reactions were quantified according to a standard scoring system, and weight loss was measured. We measured tumor control using squamous cell carcinoma (SCC) VII murine squamous cell flank tumors (50-125 mm(3)) treated with the same regimens used in the mouth irradiation model. RESULTS: We found that dFdCyd delivered 800 mg/kg once weekly or 150 mg/kg twice weekly caused similar (and maximal tolerable) weight loss; therefore these regimens were chosen to test which schedule produced more acute lip radiosensitization. Twice-weekly dFdCyd + RT was somewhat more toxic by weight loss (800 mg/kg once weekly: 11.9%; 150 mg/kg twice weekly: 17.7%; p = 0.09). To assess therapeutic index, we treated SCC VII flank tumors with RT combined with isotoxic drug/RT regimens (dFdCyd 800 mg/kg once weekly or 100 mg/kg twice weekly). Tumors treated with twice-weekly dFdCyd + RT were significantly smaller than tumors treated with once-weekly drug + RT at 28 days from the start of treatment (p < 0.03). CONCLUSIONS: These findings demonstrate that equitoxic once- versus twice-weekly dFdCyd regimens cause differing levels of oral mucosal radiosensitization. This would suggest that each radiation-dFdCyd schedule will require its own dFdCyd dose escalation trial (which cannot be determined by the maximum tolerated dose (MTD) for dFdCyd alone using that schedule). In addition, our findings suggest that for head and neck cancers twice weekly dFdCyd may have a higher therapeutic index compared with once-weekly dFdCyd when combined with daily RT. PMID- 10837966 TI - X-rays vs. carbon-ion tumor therapy: cytogenetic damage in lymphocytes. AB - PURPOSE: To measure chromosomal aberrations in peripheral blood lymphocytes from cancer patients treated with X-rays or carbon ions (C-ions). METHODS AND MATERIALS: Blood samples from patients diagnosed for esophageal or uterine cervical cancer were obtained before, during, and at the end of the radiation treatment. The novel technique of interphase chromosome painting was used to detect aberrations in prematurely condensed chromosomes 2 and 4. The fraction of aberrant lymphocytes was measured as a function of the dose to the tumor volume. For comparison, blood samples were also exposed in vitro to X-rays or to carbon ions accelerated at the HIMAC. RESULTS: C-ions were more efficient than X-rays in the induction of chromosomal aberrations in vitro. In patients with similar pathologies, tumor positions, and radiation field sizes, however, C-ions induced a lower fraction of aberrant lymphocytes than X-rays during the treatment. The initial slope of the dose-response curve for the induction of chromosomal aberrations during the treatment was correlated to the relative decrease in the number of white blood cells and lymphocytes during the treatment. CONCLUSION: C ions induce a lower level of cytogenetic damage in lymphocytes than X-rays, reducing the risk of bone marrow morbidity. PMID- 10837968 TI - Preclinical studies with the photon radiosurgery system (PRS). AB - PURPOSE: Determine the radiobiological effectiveness (RBE) for low-energy X-rays (average energy of 23 KeV) produced by the Photon Radiosurgery System (PRS). METHODS AND MATERIALS: RBE values were assessed by comparison with survival data obtained for cells irradiated with either low-energy X-rays from a GE Maxitron 100 machine or high-energy photons from a clinically used Varian 6 MV LINAC. The output of the GE and PRS sources was determined using Baldwin-Farmer and Markus thin window ionization chambers calibrated with 50 kVp X-rays and cross-checked against figures supplied by Photoelectron Corporation. The dose-rate for the PRS was 1.2 Gy/min at a distance of 35 mm with a field flatness of +/-2%. RESULTS: The RBE for the PRS low-energy X-ray source (at 1-mm depth) was greater than either the GE or Varian machines and varied with cell survival. For Chinese hamster ovary (CHO) cells, the PRS was 1.25 and 3.3 times more effective than 90 kVp X-rays and 6 MeV photons at 0.5% cell survival, respectively; by comparison, the PRS was 1.2 and 1.9 times more effective at 0.05% cell survival, respectively. Similar RBE values of 1.4 and 1.2 were obtained for human U373 and T98 glioblastoma cells grown in vitro irradiated with the PRS or GE sources, respectively. Other studies showed that the RBE for the PRS low-energy X-ray source increased with depth. The RBEs for the PRS source at 1-mm and 4-mm depth were 1.2 and 2.5 (0.5% survival) and 1. 2 and 1.9 (0.05% survival). CONCLUSIONS: The biological and physical properties of the PRS low-energy X-rays offer, under the right conditions, a significant advantage for patient treatment over conventional external beam, stereotactic, or brachytherapy treatment. PMID- 10837967 TI - Combined effects of tirapazamine and mild hyperthermia on anti-angiogenic agent (TNP-470) treated tumors-reference to the effect on intratumor quiescent cells. AB - PURPOSE: To evaluate the efficacy of the use of tirapazamine (TPZ), especially combined with mild hyperthermia (40 degrees C, 60 min), in the treatment of solid tumors following an anti-angiogenic treatment with TNP-470. In addition, we assessed the effect of TPZ and/or mild hyperthermia (MHT) combined with conventional radiotherapy or chemotherapy on TNP-470 treated tumors. MATERIALS AND METHODS: C3H/He mice bearing SCC VII tumors subcutaneously received TNP-470 at two doses of 100 mg/kg after tumor cell inoculation. At the same time, the tumor-bearing mice received 5-bromo-2'-deoxyuridine (BrdU) continuously for 5 days via implanted mini-osmotic pumps to label all proliferating (P) cells. The mice then received TPZ administration combined with or without MHT, gamma-ray irradiation combined with or without TPZ and/or MHT, or cisplatin injection with or without TPZ and/or MHT. Another group of mice received a series of test doses of gamma-rays while alive or after being killed to obtain hypoxic fractions (HFs) in the tumors at various time points after the above-mentioned cytotoxic treatment point. After each treatment, the tumors were excised, minced, and trypsinized. The tumor cell suspensions thus obtained were incubated with cytochalasin-B (a cytokinesis blocker), and the micronucleus (MN) frequency in cells without BrdU labeling (or quiescent [Q] cells) was determined using immunofluorescence staining for BrdU. The MN frequency in the total (P + Q) tumor cells was determined from the tumors that were not pretreated with BrdU. For the measurement of the HFs, the MN frequency of BrdU-unlabeled cells was then used to calculate the surviving fraction of the unlabeled cells from the regression line for the relationship between the MN frequency and the surviving fraction of total tumor cells. RESULTS: TPZ administration combined with TNP-470 treatment and MHT increased the MN frequency more markedly than treatment with TPZ alone, and this tendency was more remarkable in Q cells than total cells. In both total and Q cells, combined treatment with TPZ and MHT produced significant increases in MN frequencies whether gamma-rays were delivered to TNP-470 treated tumors or cisplatin was injected into the TNP-470 administered mice. Although not significantly, the HFs of total and Q cell populations within solid tumors increased after TNP-470 treatment. CONCLUSION: Combined treatment with TPZ and MHT, whether other cytotoxic treatments such as gamma-ray irradiation or chemotherapy using cisplatin were combined or not, was useful for sensitizing tumor cells in vivo including Q cells even after TNP-470 treatment. PMID- 10837969 TI - Dosimetric and volumetric criteria for selecting a source activity and a source type ((125)I or (103)Pd) in the presence of irregular seed placement in permanent prostate implants. AB - PURPOSE: The dosimetric merit of a permanent prostate implant relies on two factors: the quality of the plan itself, and the fidelity of its implementation. The former factor depends on source type and on source strength, while the latter is a combination of skill and experience. The purpose of this study is to offer criteria by which to select a source type ((125)I or (103)Pd) and activity. METHODS AND MATERIALS: Given a prescription dose and potential seed positions along needles, treatment plans were designed for a number of seed types and activities, specifically for (125)I with activities ranging from 0.3 to 0.7 mCi, and for (103)Pd with activities in the range of 0.8 to 1.6 mCi. To avoid human planner bias, an automated computerized planning system based on integer programming was used to obtain optimal seed configurations for each seed type and activity. To simulate the effect of seed-placement inaccuracies, random seed displacement "errors" were generated for all plans. The displacement errors were assumed to be uniformly distributed within a cube with side equal to 2sigma. The resulting treatment plans were assessed using two volumetric and two dosimetric indices. RESULTS: For (125)I implants a coverage index (CI) of 98.5% or higher can be achieved for all activities (CI is the fraction of the target volume receiving the prescribed or larger dose). The external volume index (EI) (i.e., the amount of healthy tissue, as percentage of the target volume, receiving the prescribed or larger dose) increases from 13.9% to 20% as the activity increases from 0.3 to 0.7 mCi. For implants using (103)Pd, the external volume index increases from 10. 2% to 13.9% whenever CI exceeds 98.5%. Volumetric and dosimetric indices (coverage index, external volume index, D90, and D80) are all sensitive to seed displacement, although the activity dependence of these indices is more pronounced for (125)I than for (103)Pd implants. CONCLUSIONS: For both isotopes, the lower activities studied systematically result in lower EIs. If seeds can be placed within approximately 0.5 cm of their intended position (103)Pd should be preferred because its EI is lower than that of (125)I. For all activities the coverage indices and D90 are within the required range. If seed placement uncertainties are larger than 0.5 cm, (125)I provides slightly better target coverage; however, in terms of external volume (healthy tissue) covered, (103)Pd is superior to (125)I. PMID- 10837970 TI - Dosimetric evaluation of abutted fields using asymmetric collimators for treatment of head and neck. AB - PURPOSE: The objective of this study was to reevaluate the dose nonuniformity of abutted fields defined using asymmetric collimators and one isocenter for treatment of the head and neck region. METHODS AND MATERIALS: Bilateral parallel opposed fields abutted to the anterior field at one isocenter were implemented in the treatment of head and neck. The effect of digital display tolerance can produce dose nonuniformity at the junction of the abutted fields. The amount of dose nonuniformity was quantified using both mathematical summation of dose profiles and by direct measurement of doses at the junction of the two abutted fields. The dose nonuniformity was obtained by irradiating the superior part of a film using bilateral parallel-opposed fields and the inferior part by an anterior field with a gap or an overlap. Dose profiles were taken at the depth of maximum dose for the anterior field across the abutted fields. The dose nonuniformity was determined for the case where the asymmetric jaw was set at -2 mm, -1 mm, 0, +1 mm, and +2 mm from the beam central axis. RESULTS: The dose at the junction increases systematically as the abutment of the fields changes from a gap to an overlap. The dose nonuniformity with 1-mm gap and 1-mm overlap is about 15% underdose and overdose, respectively. CONCLUSION: Imperfect abutment of split fields due to digital display tolerance (+/-1 mm) of asymmetric collimator can cause an underdose or overdose of 15% of the delivered dose. PMID- 10837971 TI - Setup error in radiotherapy: on-line correction using electronic kilovoltage and megavoltage radiographs. AB - PURPOSE: We hypothesize that the difference in image quality between the traditional kilovoltage (kV) prescription radiographs and megavoltage (MV) treatment radiographs is a major factor hindering our ability to accurately measure, thus correct, setup error in radiation therapy. The objective of this work is to study the accuracy of on-line correction of setup errors achievable using either kV- or MV-localization (i.e., open-field) radiographs. METHODS AND MATERIALS: Using a gantry mounted kV and MV dual-beam imaging system, the accuracy of on-line measurement and correction of setup error using electronic kV and MV-localization images was examined based on anthropomorphic phantom and patient imaging studies. For the phantom study, the user's ability to accurately detect known translational shifts was analyzed. The clinical study included 14 patients with disease in the head and neck, thoracic, and pelvic regions. For each patient, 4 orthogonal kV radiographs acquired during treatment simulation from the right lateral, anterior-to-posterior, left lateral, and posterior-to anterior directions were employed as reference prescription images. Two dimensional (2D) anatomic templates were defined on each of the 4 reference images. On each treatment day, after positioning the patient for treatment, 4 orthogonal electronic localization images were acquired with both kV and 6-MV photon beams. On alternate weeks, setup errors were determined from either the kV or MV-localization images but not both. Setup error was determined by aligning each 2D template with the anatomic information on the corresponding localization image, ignoring rotational and nonrigid variations. For each set of 4 orthogonal images, the results from template alignments were averaged. Based on the results from the phantom study and a parallel study of the inter- and intraobserver template alignment variability, a threshold for minimum correction was set at 2 mm in any direction. Setup correction was applied by translating the treatment couch in the lateral, superior-to-inferior and vertical directions only. During treatment, kV open-field images were acquired for off-line treatment verification and analysis. Each patient study spanned 2-6 weeks. The 14 patient studies were completed with 8248 electronic images acquired and analyzed. RESULTS: Results from the phantom studies showed that the users were able to detect the applied translational shift to better than 2 mm, and mostly to within 1 mm. The intraobserver variability of template alignment was on the order of 1 mm using a sample of either MV or kV patient images. The difference between using MV or kV images was significant for only a few cases. However in most cases, interobserver alignment variability was larger when using MV images than kV. For on-line setup correction, the study procedure added 10 min. to conventional treatment time. Setup variation measured with either kV- or MV-localization images was similar. The initial magnitude of setup error was appreciable, with a mean displacement of about 6.6 +/- 2.4 mm for the 14 patients. On-line correction using either kV- or MV-localization images improved setup accuracy. Over all study patients, setup errors occurred with standard deviations greater than 2 mm in any direction with a frequency of 48% before correction, and were reduced to 16% after correction. On average, kV image-based correction reduced radial setup variation to 2.6 +/- 1.6 mm compared to the 3.3 +/- 1.8 mm attained using MV images. The difference detected between the kV and MV data was not statistically significant when averaged over all patients. However, for on-line corrections in the neck and thoracic regions, using kV-localization images reduced setup error significantly more than using MV images. CONCLUSIONS: In our anatomic template alignment study, interobserver variability was smaller using kV images than MV images. Intraobserver variability was smaller for alignments on kV images PMID- 10837972 TI - The impact of dose-specification policies upon nominal radiation dose received by breast tissue in the conservation treatment of breast cancer. AB - PURPOSE: In the context of breast conservation treatment, absorbed dose is influenced by (1) prescribed nominal dose, and (2) dose-specification characteristics employed. Breast doses are generally specified either at tangent isocenter, varying anatomical points within the breast, or at isodoses varying from 90% to 100%. Boost doses are generally specified at 80-100%. METHODS: An idealized axial slice of breast tissue at central axis is presented. Assuming varying dose-specification characteristics, absorbed doses are normalized and compared to those received by nominal prescriptions of 46 Gy to the breast and 20 Gy to the boost volume, both specified at 100%. RESULTS: Absorbed doses vary from the normalized total of 66 Gy (with specification of breast and boost at 100%) in gradations up to a maximum of 76.11 Gy (when breast dose is specified at the 90% isodose and boost dose at 80%), a 13.3% difference. CONCLUSION: The impact of dose specification is largely ignored in the breast irradiation literature and unappreciated in clinical practice. Its impact, however, is illustrated as dwarfing modest nominal dose escalations commonly recommended and prescribed among margin compromised patients. Progress in delineation of a dose-response relationship for treatment of breast cancer requires consensus as to dose specification. Arguments are offered that ICRU Report 50 dose-specification standards, as verified for reproducibility by the EORTC (22881/10882) trial group, constitutes the best data source currently available from which dose specification consensus may be reached (1, 2). Dose to PTV(1) (whole breast plus 1- to 2-cm margin) should be specified at the tangent beam intersection on the central plane or, where such point is irrelevant, at two-thirds distance from dorsal beam edge to skin along the perpendicular breast bisector. Where irradiated via electrons, dose to the boost PTV(2) (lumpectomy cavity plus 1- to 3-cm margins) should be specified at 90%. Electron energy sufficient to provide 85% isodose coverage to all aspects of PTV(2) is recommended. PMID- 10837974 TI - Induction of parturition with prostaglandin f2 alpha as a possible model to study impaired reproductive performance in the dairy cow. AB - Parturitions were induced in five cows, 2 weeks before term using prostaglandin (PG) F(2alpha). Two i.m. injections were performed with an interval of 24 h. All cows calved within 5 days (average 2.7 days) after the first injection of PGF(2alpha). Out of five cows, four had retained fetal membranes (RFM). Each animal was sampled for bacteriological examination using uterine biopsies twice a week during 42 days postpartum (PP). Jugular vein blood samples were withdrawn for PGF(2alpha)-metabolite and progesterone analyses five times per day during the first week PP and eight times per 24 h during the 2nd and 3rd weeks PP. From the 4th week, the sampling interval was reduced back to five times per day. From the 5th week PP, the sampling was reduced to two times per day and sampling was terminated after day 46 PP. Only morning samples were used for progesterone analyses. From day 10 PP, ultrasonography (US) was performed every 3rd day until day 39 PP for detection of ovarian activity and follicular dynamics. The highest incidence of bacteriological species was found during the first 3 weeks PP. After the 5th week of collection, all animals were free from bacteria. The species of bacteria found were Arcanobacterium (Actinomyces) pyogenes, Escherichia coli, alpha-hemolytic streptococcae and Pasteurella multocida. Immediately after parturition, very high levels of the PG-metabolite were seen in all animals, with a sharp decrease to line of significance around days 9-12 PP. Small increases above the line of significance were detected up to day 27 PP in cows with RFM, and after that time the levels were considered to be at baseline. Low levels of progesterone were seen in four animals during the whole experimental time. In one animal, an increase was seen on day 43 PP, which was maintained until the end of the experimental period on day 46 PP. Based on US, follicular waves were detected in all animals during the experimental period. In three animals, three non ovulatory follicular waves were detected and in two animals, four non-ovulatory follicular waves were detected during 39 days of ultrasound sessions. Based on progesterone levels, only one animal was considered to have ovulated around day 40 PP. Results from the present study indicate that reproductive performance of cows after PG-induced parturitions differs from those of spontaneous cases of RFM. Differences regarding the resumption of ovarian activity were also observed between previous studies of dexamethasone-induced parturitions and the present study. PMID- 10837975 TI - Local versus systemic effects of exogenous estradiol-17 beta on ovarian follicular dynamics in heifers with progestogen implants. AB - Two experiments were designed to determine if the suppressive effect of estradiol treatment on ovarian follicles in progestogen-implanted heifers is mediated directly at the ovary or systemically, at a higher level. The purpose of Experiment 1 was to determine a minimal effective dose of estradiol-17beta (E 17beta) that would induce follicle regression in progestogen-implanted heifers. Beef heifers were implanted with progestogen on Day 2 (Day 0=ovulation) and were assigned randomly to five groups: control (sesame seed oil, n=9); 0. 1 mg of E 17beta (n=8); 0.5 mg of E-17beta (n=8); 1 mg of E-17beta (n=8); or 5 mg of E 17beta (n=8) by intramuscular (im) injection on Day 3. Treatment with 5 and 1 mg of E-17beta resulted in smaller (P<0.05) day-to-day diameter profiles of the dominant follicle compared with controls, whereas 0.1 mg of E-17beta did not have an apparent effect on follicle growth. The effect of a dose of 0.5 mg was intermediate and tended (P<0.06) to result in a smaller diameter profile of the dominant follicle compared with control heifers. Experiment 2 was designed to utilize a subminimal dose of E-17beta (0.1 mg), locally, to determine whether estradiol treatment induces follicle regression through a direct action on the ovary. Beef heifers received a progestogen ear implant on Day 2 and were assigned randomly to five groups on Day 3: control (sesame seed oil, n=8); 5 mg of E 17beta im (n=8); 0.1 mg of E-17beta im (n=8); 0.1 mg of E-17beta given into the wall of the uterus, near the tip of the horn ipsilateral to the dominant follicle (intrauterine (iu), n=8); or 0.1 mg of E-17beta given into the stroma of the ovary, immediately adjacent to the dominant follicle (intraovarian (io), n=6). Local (iu and io) treatments were given via a transvaginal ultrasound-guided needle injection. Treatment with 5 mg of E-17beta im resulted in suppression of the dominant follicle of the first follicular wave and early emergence of the second follicular wave (P<0.05). Diameter profiles of the dominant follicle in heifers treated with 0.1 mg im or 0.1 mg iu differed from those of control heifers on Day 5, whereas diameter profiles of the dominant follicle in heifers treated with 0.1 mg io did not differ from the controls. Daily changes in diameter of the dominant follicle did not differ among the three groups treated with 0.1 mg of E-17beta (im, iu and io). Hourly changes in circulating concentrations of FSH and LH were not detected following estradiol treatment either before or after the results were combined for all estradiol-treated groups. Results are supportive of the hypothesis that the suppressive effect of estradiol in cattle is exerted indirectly through a systemic route rather than directly at the ovary. Although low plasma concentrations of FSH and LH were not detected, systemic treatments with high E-17beta dosages resulted in follicular suppression whereas local treatments with subminimal dosages, within the ovary bearing the dominant follicle, were without effect. PMID- 10837976 TI - Effectiveness of lipopolysaccharide as an intrauterine immunomodulator in curing bacterial endometritis in repeat breeding cross-bred cows. AB - Antibiotics are usually used to combat microbial infections of the uterus, responsible for hindering establishment of pregnancy in cross-bred cows. The major disadvantages of antibiotics are: development of bacterial resistance, high costs and diminishing uterine defense mechanisms (UDM). As an alternative therapy, intrauterine application of Escherichia coli Lipopolysaccharide (E. coli LPS) as a uterine defense stimulator was used in this study in confirmed clinical cases of repeat breeding associated with bacterial endometritis. In the treated group (n=12), on the day of estrus, 100 microg of E. coli LPS dissolved in 30-ml sterile phosphate buffer saline (PBS) was infused intrauterine; while in the control group (n=12), only 30 ml of PBS was infused. Six-hour post-treatment, in the treatment group uterine washings showed a 100-fold increase in the total leucocytic count (TLC). Out of the cellular contents, more than 80% of the cells were recognised as neutrophils; above 60% were alive and their phagocytic activity was five bacteria/neutrophil. Such a cellular response was maintained until 72-h post-treatment. At the subsequent estrus period, the cervicovaginal mucus (CVM) became clear in 9 out of 12 cows (75%) and showed no bacterial growth. In the control group, similar micro-organisms were present in CVM of all the 12 cows before and after the PBS infusions. During the subsequent estrus, all nine cows with sterile CVM in the treatment group conceived while only one cow conceived from the control group. It was concluded that, administration of intrauterine E. coli LPS as single infusion in cows with bacterial endometritis stimulated UDM and cleared the infection within one estrous cycle, and thereby restoring fertility. PMID- 10837977 TI - Effect of stress-like concentrations of cortisol on the feedback potency of oestradiol in orchidectomized sheep. AB - The effect of stress-like concentrations of cortisol on oestradiol-induced change in LH secretion and GnRH receptor expression was evaluated in orchidectomized sheep (wethers). Twenty-four wethers were assigned at random to one of the four treatment groups in a 2x2 factorial design (n=6 wethers/group). Wethers received cortisol (90 microg/kg/h; groups 2 and 4) or a comparable volume of cortisol delivery vehicle (groups 1 and 3) by continuous infusion for 48 h. During the final 24 h of infusion, wethers received oestradiol (6 ng/kg/h; groups 3 and 4) or oestradiol delivery vehicle (groups 1 and 2). The pattern of LH secretion was assessed during a 3-h period of intensive blood collection beginning 21 h after initiation of oestradiol infusion. Although neither cortisol nor oestradiol alone affected (P>0.05) mean serum concentration of LH or LH pulse frequency, serum LH and the frequency of secretory episodes of LH were significantly reduced (P<0.05) in wethers receiving cortisol and oestradiol in combination. Anterior pituitary tissue was collected at the end of the infusion period. Oestradiol increased (P<0.05) tissue concentrations of GnRH receptor and GnRH receptor mRNA. Although cortisol alone did not affect (P>0.05) basal concentrations of receptor or receptor mRNA, the magnitude of oestradiol-induced increase in GnRH receptor and GnRH receptor mRNA was significantly reduced in wethers receiving cortisol and oestradiol concurrently. Conversely, steady-state concentrations of mRNA encoding the LHbeta and FSHbeta subunits were increased (P<0.05) in wethers receiving cortisol. These observations demonstrate that stress-like concentrations of cortisol act in concert with oestradiol to suppress LH secretion. In addition, cortisol blocks oestradiol-dependent increase in pituitary tissue concentrations of GnRH receptor and GnRH receptor mRNA. PMID- 10837978 TI - Real-time ultrasonography of ovaries and breeding of the one-humped camel (Camelus dromedarius) during the early postpartum period. AB - Ovaries of 17 adult, pleuriparous, and lactating one-humped she-camels (Camelus dromedarius) were examined per rectum for uterine involution and for presence of follicles (>/=1.0 cm diameter) by real-time ultrasonography at the National Research Centre on Camel at Bikaner, India at 30, 45, 60, 75, and 90 days postpartum. Involution was completed from 25 to 30 days postpartum and follicles (>/=1.0 cm diameter) could be found in only nine camels (52.7%) from 34 to 70 days postpartum. These nine camels were mated with virile studs. Four conceived and were confirmed pregnant at 60 days. PMID- 10837979 TI - Effects of post-ovulatory food deprivation on the hormonal profiles, activity of the oviduct and ova transport in sows. AB - The objective of the present study was to investigate the effect of post ovulatory food deprivation on the hormonal profiles and consequently on the activity of the oviduct and ova transport in sows. Sows were randomly allocated to the control (C-group, n=6) or fasted (F-group, n=5) group. The F-group sows were fasted for four meals starting with the morning meal after detection of ovulation in the second oestrus after weaning. Ovulation was checked by transrectal ultrasonography. Blood was collected for the analyses of progesterone, oestradiol-17beta, prostaglandin F(2 approximately ) metabolite, insulin, free fatty acids and triglycerides. Oviductal isthmic motility was monitored on Polyview before and after ovulation until the time of slaughter. After slaughter, the isthmus opposite the side with transducer was divided into three equal segments and flushed separately and a third of the uterine horn part from the utero-tubal-junction (UTJ) was also flushed. A high proportion of ova in the F-group was found in the first and second parts of the isthmus. In the C group, a high proportion of ova was found in the third part of the isthmus and the uterus. The mean isthmic pressure in the C-group decreased significantly (P<0.05) during the period immediately after ovulation while in the F-group mean pressure remained unchanged. The frequency of phasic pressure fluctuations were significantly (P<0.05) higher in the F- than in the C-group 13 to 24 h after ovulation. No significant differences in progesterone concentrations were seen between the two groups of sows. Prostaglandin metabolite levels were significantly (P<0.05) higher in the F-group than in the C-group. Oestradiol 17beta levels significantly (p<0.05) decreased earlier in the F- than in the C group. Serum insulin levels were significantly (p=0.05) lower in the F- than in the C-group while free fatty acids were significantly (p<0.01) higher in the F- than in the C-group. There were no significant differences in the serum levels of triglycerides between the F- and the C-group. Therefore, it can be concluded in the present study that food deprivation is associated with changes in the hormonal profiles, activity of the oviduct and a delay of ova transport in sows. PMID- 10837980 TI - The use of transmigration and spermac stain to evaluate epididymal cat spermatozoa. AB - Epididymal spermatozoa of domestic cats were diluted with TEST medium and frozen. The parameters - estimated percentage of motile spermatozoa, concentration of spermatozoa, cell morphology and transmigration rate (TMR) - were evaluated before freezing and after thawing. Transmigration is a new method to measure the percentage of spermatozoa that consistently move forward, and has not been investigated with cat spermatozoa until now. Estimated percentage of motile spermatozoa averaged 65%, TMR was 76%, concentration of spermatozoa was 30,000 microl(-1) and the incidence of morphologically abnormal spermatozoa averaged 58% before freezing. After thawing, the estimated number of motile spermatozoa declined by 22%, but TMR remained at 76%. The TMR did not correlate with estimated motility but mostly was higher than the latter, which is postulated to be caused by the mobilizing effect of the countercurrent in the transmigration apparature. The estimated percentage of motile cells in the target chamber of the transmigration apparature was improved by using phosphate-buffered saline (PBS) as transmigration medium. Morphology was assessed both after fixation of spermatozoa in Hancock solution and after staining of smears with Spermac. Spermac did not stain all protoplasmic droplets but proved to be more suitable for the routine examination of acrosomal morphology after thawing. PMID- 10837981 TI - Sperm transport in the female reproductive tract of the brushtail possum, Trichosurus vulpecula, following superovulation and artificial insemination. AB - This study investigated sperm transport following superovulation and artificial insemination (AI) in the common brushtail possum, Trichosurus vulpecula. Females were superovulated by treatment with 15 IU pregnant mare serum gonadotrophin (PMSG) then 4 mg luteinizing hormone (LH) 78 h later. Inseminations were performed 27 h after LH (4 million motile spermatozoa/uterus). At 1.5, 3, 6, 9 and 12 h after AI (n=5 per group), females were euthanised and reproductive tracts removed for examination and flushed for sperm. No ovulations had occurred by 1.5 h, but 20% of animals had ovulated by 3 or 6 h, and 80% by 9 or 12 h. The mean numbers of spermatozoa recovered ranged from 249 to 275x10(3) in the uterus; 16-51x10(3) in the isthmus; 8-11x10(3) in the middle segment; and 6-16x10(3) in the ampulla at 1.5, 3 and 6 h after AI. Sperm numbers in all regions decreased at later times (P<0.05) except the isthmus, where 100x10(3) sperm were recovered by 12 h. Highly motile thumbtack sperm (a putative indicator of capacitation in marsupials), were recovered from the isthmus (20%), middle segment (50%) and ampulla (90%) at all sampling times, but not from the uterus. The epithelium of the oviduct segments contained mucus-secreting and ciliated cells and peak secretory activity was observed in the ampulla at 6 h. At 3, 6 and 12 h, many spermatozoa were found in epithelial folds within the isthmus. The present study has provided basic information on sperm transport and storage events within the female reproductive tract of T. vulpecula following superovulation and AI. It is concluded that this model may be useful to better understand pre-fertilization sperm maturation events in the possum, which could facilitate the development of IVF technology. PMID- 10837982 TI - Medicinal plants from nepal: evaluation as inhibitors of leukotriene biosynthesis. AB - The methanolic extracts of 25 different Nepalese medicinal plants were tested for their activity to inhibit the biosynthesis of leukotriene B(4) in bovine polymorphonuclear leukocytes. The selected indigenous plants are used in traditional herb remedies to treat inflammatory diseases such as asthma, bronchitis, rheumatism, and skin disorders presumed to be mediated by leukotrienes. The leaves of Zanthoxylum nepalensis were shown to be the most potent inhibitor with an IC(50) value of 11 microgram/ml. The extracts obtained from Astercantha longifolia and Hedychium ellipticum also exhibited potent inhibitory action with IC(50) values of 20 and 22 microgram/ml, respectively. PMID- 10837983 TI - Oldenlandia affinis (R&S) DC. A plant containing uteroactive peptides used in African traditional medicine. AB - A review of the geographical distribution, clinical use, biological activity and phytochemistry of Oldenlandia affinis (R&S) DC. is presented. During an inventory of medicinal plants in northern Congo/Brazzaville and south-western Central African Republic in 1962, 196 different species were registered, one of which was O. affinis used for the facilitation of childbirth. A medical team working in Luluabourg (Kananga) in Congo during the troubled period in 1960, discovered also the traditional use of the same plant as an oxitocic agent during labour. The plant was collected and the uterotonic substances isolated. Cyclic peptides (called Kalata-peptides) were described, and the main peptide, B1, was subjected to pharmacological and chemical investigations. Later the three-dimensional structure of the peptide was determined. Similar cyclic peptides have been isolated also from other plants in the Rubiaceae family like Chassalia pasvifoloia and Psychotria longipes, and from Viola species: Viola tricolor L. and Viola arvensis Murray. Some of these peptides, included Kalata-peptide B1, have been shown to hold antimicrobial activity. They have recently been synthesized, and they may represent a starting point for the design of new peptide antibiotics. PMID- 10837984 TI - Effect of papaya latex extract on gravid and non-gravid rat uterine preparations in vitro. AB - In search of uterotonic principles, papaya (Carica papaya, Caricaceae) latex extract (PLE) was tested on rat uterine preparations in vitro at various stages of the estrous cycle and gestation periods. Rat uterine contractile activity was remarkably increased by different doses of PLE in proestrus and estrus stages compared to metestrus and diestrus stages of the estrous cycles. The maximum contractile activity of the uterus was observed at the later stages of pregnancy which correspond with the peak level of estrogen in the plasma. A direct dose dependent spasmodic action with increased frequency and amplitude was observed with PLE in all non-gravid uterine preparations. Pretreatment of the tissue with phenoxybenzamine (PB) non-competitively blocked the effect of PLE. Blocking of the 5-HT receptors with methysergide partially blocked the excitatory response to PLE. Pretreating the tissue with Indomethacin, a cyclo-oxygenase inhibitor, had no effect on the response to PLE. The release of PLE induced mast cell degranulation and subsequent release of heparin, biogenic amines or prostaglandins (PGs) was ruled out by pretreating the tissue with sodium cromoglycate, a mast cell stabilizer. Pure papain induced uterine contractions were not sustained for a longer period and at higher concentrations the receptor proteins were affected by the enzymatic action of papain. From this study it is evident that the crude papaya latex contain a uterotonic principle which might be a combination of enzymes, alkaloids and other substances which can evoke sustained contraction of the uterus acting mainly on the alpha adrenergic receptor population of the uterus at different stages. PMID- 10837985 TI - Studies on anti-complementary activity of extracts and isolated flavones from Ligustrum vulgare and Phillyrea latifolia leaves (Oleaceae). AB - Polar fractions and flavones isolated from methanolic extracts of the leaves of Ligustrum vulgare and Phillyrea latifolia (Oleaceae), whose popular use as an anti-inflammatory is well-known in Mediterranean historical medicine and ethnobotany, showed significant in vitro complement inhibiting effect on the classical pathway of the complement system. Among the isolated flavonoidic structures, apigenin-7-O-glucoside, apigenin-7-O-rutinoside, luteolin-4'-O glucoside, luteolin-7-O-glucoside and ligustroflavone presented remarkable activity. PMID- 10837986 TI - Anti-ulcerogenic effects of Spartium junceum flowers on in vivo test models in rats. AB - Flowers of Spartium junceum L. (Fabaceae) are used for the treatment of gastric ulcers in Turkish folk medicine. Through bioassay-guided fractionation using chemical and chromatographical means and water immersion and restraint-induced stress ulcer model in rats, a saponin fraction was determined as the potent anti ulcerogenic ingredient. The active fraction was also highly effective in preventing ethanol- and pyloric ligation-induced gastric lesions as well as inhibiting gastric secretion volume, gastric pH and titratable acidity, but did not affect the hexosamine content of the gastric mucosa. A novel oleanen-type triterpenic saponin, named as spartitrioside, was isolated as the active principle by using chromatographical separation techniques. PMID- 10837988 TI - Medicinal plants and food medicines in the folk traditions of the upper Lucca Province, Italy. AB - An ethnopharmacobotanical survey of the medicinal plants and food medicines of the northern part of Lucca Province, north-west Tuscany, central Italy, was carried out. The geographical isolation of this area has permitted the survival of a rich folk phytotherapy involving medicinal herbs and also vegetable resources used by locals as food medicine. Among these are the uncommon use of Ballota nigra leaves as a trophic protective; the use of Lilium candidum bulbs as an antiviral to treat shingles (Herpes zoster); Parmelia sp. as a cholagogue; Crocus napolitanus flowers as antiseptic; Prunus laurocerasus drupes as a hypotensive; and the consumption of chestnut flour polenta cooked with new wine as bechic. Many wild gathered greens are eaten raw in salads, or in boiled mixtures, as 'blood cleansing' and 'intestine cleansing' agents. Of particular interest is the persistence of the archaic use of Bryonia dioica root against sciatica, and the use of ritual plant therapeuticals as good omens, or against the 'evil eye.' Over 120 species represent the heritage of the local folk pharmacopoeia in upper Garfagnana. Anthropological and ethnopharmacological considerations of the collected data are also discussed. PMID- 10837987 TI - Tecoma sambucifolia: anti-inflammatory and antinociceptive activities, and 'in vitro' toxicity of extracts of the 'huarumo' of peruvian incas. AB - Aqueous and alcoholic extracts of pods and flowers of Tecoma sambucifolia H.B.K. (Bignoniaceae) ('huarumo') were analysed to determine their anti-inflammatory activity (carrageenan-induced edema test), antinociceptive activity (acetic acid writhing test) and 'in vitro' toxicity in Chinese hamster ovary cells, human hepatome cells and human larynx epidermal carcinoma cells. The cytotoxic effects of both extracts were evaluated by two endpoint systems: neutral red uptake assay and tetrazolium assay. The results showed that all extracts have anti inflammatory and antinociceptive activity, but the highest potency is that of the alcoholic extracts. There were significant differences in cytotoxicity between extracts and among the response of cells to them. The highest cytotoxicity was noted with the alcoholic extract, and the human hepatome cell line was the most sensitive, especially to the alcoholic extract of flowers. The aqueous pod extract appeared to have the best pharmaco-toxicological profile, since it provided a significant reduction of both pain and inflammation together with the lowest cytotoxicity. PMID- 10837989 TI - Hypnotic, anticonvulsant and muscle relaxant effects of Rubus brasiliensis. Involvement of GABA(A)-system. AB - Rubus brasiliensis hexanic fraction induced anxiolysis in rodents, which was reversed by flumazenil, a specific GABA(A)-benzodiazepine receptor antagonist (Nogueira et al., 1998a,b). Then, we investigated if this hexanic fraction was able to induce hypnotic, anticonvulsant and muscle relaxant effects, and the involvement of GABA(A)-system. The hexanic fraction (50, 100, 150 and 300 mg/kg, vo) was administered to male Swiss mice, 30 min before the tests. Only the dose of 300 mg/kg of this fraction decreased the latency and increased sleeping time in the barbituric-hypnosis test (sodium pentobarbital, 30 mg/kg, ip), prevented the pentylenetetrazol seizures (70 mg/kg, ip) and induced muscle relaxant (inclined plane) in 100% of animals. These effects were reversed by flumazenil (3 mg/kg, ip). IN CONCLUSION: (1) R. brasiliensis hexanic fraction induced hypnotic, anticonvulsant and muscle relaxant effects, in mice, and the GABA(A) benzodiazepine receptor may play an important role in the effects of this fraction; (2) it is strongly suggested that this fraction contains a benzodiazepine-like principle. PMID- 10837990 TI - Traditional herbal drugs of southern Uganda, I. AB - One-hundred four plant species used medicinally by herbalists from three southern Ugandan tribes were collected and identified. The collection includes a large portion of the materia medica of the Abayanda of the southwest region, as well as species used by herbalists of the Baganda and Bakiga Tribes. Literature searches were performed in preparation for further collections, and for collaborative laboratory validation of in vitro antimicrobial activity. Literature data provide support for ethnomedical claims for a number of species used in Uganda for disease treatment. PMID- 10837991 TI - Isolation of hypotensive compounds from Solanum sisymbriifolium Lam. AB - The crude hydroalcoholic root extract (CRE) of Solanum sisymbriifolium Lam. has formerly been shown to have hypotensive activity both in normo-and hypertensive rats. Hypotensive activity-guided fractionation of the CRE was performed in anaesthetized normotensive rats, which led to the isolation of the active principles. The intravenous (i.v.) and intraperitoneal (i.p.) values of the CRE in mice were found to be, respectively, 343 and 451 mg/kg, and no lethal effect was caused by doses up to 5.0 g/kg when administered by oral route. Depression of locomotion, increase of breathing rate and piloerection was observed in a general behavior test with doses up to 200 mg/kg i.p., and 1000 mg/kg p.o., respectively. Increase in the gastrointestinal transit was found using 0.1 g/kg, whereas at doses of 0.5 and 1 g/kg, no significant activity was observed in comparison with the control mice. Hexanic and butanolic fractions induced a remarkable hypotension in anaesthetized normotensive rats in doses of 1, 5, 7.5 and 10 mg/kg i. v. Two compounds isolated from the butanolic fraction induced a significant decrease of the blood pressure, HR, amplitude of the ECG and breathing rate when injected in a dose of 1 mg/kg i.v; and both systofic and diastolic, blood pressures were affected in a proportional mode. The hypotensive effect of the two compounds were not influenced by pretreatment with atropine and propranolol; and the pressor response to noradrenaline was not affected by any of them which suggests that neither a direct muscarinic activity, beta-adrenoceptor activation nor decrease of sympathetic vascular tone (sympatholitic activity) are probably involved in the mechanism of hypotension. The present study shows that the CRE of S. sisymbriifolium contains at least two hypotensive compounds whose characterization is under way. PMID- 10837992 TI - Effect of Punica granatum Linn. (flowers) on blood glucose level in normal and alloxan-induced diabetic rats. AB - 'Gulnar farsi', male abortive flowers of Punica granatum L., are used for the treatment of diabetes mellitus in Unani medicine. Oral administration of its aqueous-ethanolic (50%, v/v) extract led to significant blood glucose lowering effect in normal, glucose-fed hyperglycaemic and alloxan-induced diabetic rats. This effect of the extract was maximum at 400 mg/kg, b.w. PMID- 10837993 TI - Evaluation of in-vivo wound healing activity of Hypericum patulum (Family: hypericaceae) leaf extract on different wound model in rats. AB - The methanol extract of Hypericum patulum Thumb. leaves were investigated for the evaluation of their wound healing potential on different experimental models of wounds in rats. The methanol extract of leaves (HPM), in the form of an ointment with two different concentrations (5% and 10% w/w ointment of leaf extract in simple ointment base) was evaluated for wound healing potential in an excision wound model and an incision wound model in rats. Both concentrations of the methanol extract ointment showed significant responses in both the wound types tested when compared with the control group. The effect produced by the extract ointment, in terms of wound contracting ability, wound closure time, regeneration of tissues at wound site, tensile strength of the wound and histopathological characteristics were comparable to those of a standard drug nitrofurazone ointment. PMID- 10837994 TI - Antinociceptive effect of the essential oil from Cymbopogon citratus in mice. AB - The essential oil (EO) from leaves of Cymbopogon citratus increased the reaction time to thermal stimuli both after oral (25 mg/kg) and intraperitoneal (25-100 mg/kg) administration. EO (50-200 mg/kg, p.o. or i.p.) strongly inhibited the acetic acid-induced writhings in mice. In the formalin test, EO (50 and 200 mg/kg, i.p.) inhibited preferentially the second phase of the response, causing inhibitions of 100 and 48% at 200 mg/kg, i.p. and 100 mg/kg, p.o., respectively. On the other hand, the opioid antagonist naloxone blocked the central antinociceptive effect of EO, suggesting that EO acts both at peripheral and central levels. PMID- 10837995 TI - Gastric cytoprotection of bolivian medicinal plants. AB - Several extracts obtained from Bolivian medicinal plants have been evaluated for cytoprotective activity on ethanol-induced ulcer formation in rats. Preliminary results suggest, that the majority of the plants tested showed a significant activity, the aqueous extracts of Phoradendron crassifolium and Franseria artemisioides being the most active, exerting a cytoprotective activity comparable to atropine. The analysis of the chemical constituents of the extracts studied showed the presence of tanins, saponins, flavonoids and coumarins. PMID- 10837996 TI - Evaluation of antifungal activity of extracts of two Cameroonian rutaceae: Zanthoxylum leprieurii Guill. et Perr. and Zanthoxylum xanthoxyloides Waterm. AB - Aqueous-ethanol 90% extracts of leaves, roots and stem barks of Zanthoxylum leprieurii and Zanthoxylum xanthoxyloides were examined for their antifungal properties against nine fungi by dilution methods on a solid medium and in a liquid medium. Our results indicate that these extracts, to varying extents, inhibit the in vitro growth of Candida albicans, Cryptococcus neoformans and seven filamentous fungi tested. Only the extracts obtained from the roots and stem barks of Z. xanthoxyloides showed antifungal activity on the germs studied, with minimal inhibitory concentration varying, respectively, from 0.5 to 1 mg/ml for the roots and from 0.125 to 1 mg/ml for the stem barks. PMID- 10837997 TI - Screening of some Palestinian medicinal plants for antibacterial activity. AB - Antibacterial activity of organic and aqueous extracts of 15 Palestinian medicinal plants were carried against eight different species of bacteria: Bacillus subtilis, two Escherichia coli species, Staphylococcus aureus (methicillin resistant), two S. aureus (methicillin sensitive) species, Pseudomonas aeruginosa, and Enterococcus fecalis. Of the 15 plants tested, eight showed antibacterial activity. Each plant species has unique against different bacteria. The most active antibacterial plants against both gram-positive and gram-negative bacteria were Thymus vulgaris and Thymus origanium. The organic and aqueous extract from the same plants showed different activities; the organic extract showed the same or greater activity than the aqueous extract. Finally, the hole-plate diffusion method showed larger activity than the disc diffusion method. PMID- 10837998 TI - Education and counseling in cancer: the neglected case of the prostate cancer patients. PMID- 10837999 TI - Motivations, satisfaction, and information of immediate breast reconstruction following mastectomy. AB - This study evaluated patients' motivations for, and satisfaction with, the treatment and information of immediate breast reconstruction (IBR) with a silicone prosthesis. It studied satisfaction more deeply by relating it to the quality of life, body-image and sexual functioning. Seventy-three patients who received mastectomy, followed by IBR with a subpectoral silicone prosthesis, completed a self-report questionnaire concerning their motivations for, perceived advantages of and satisfaction with IBR, the information received, quality of life, body image, and sexual functioning. Despite the fact that 50% of the reconstructions resulted in complications or complaints, 70% of the women were satisfied with the reconstruction and only 12% would never choose IBR again. Satisfaction was strongly correlated with the need for information. The higher the patient's expectations, the higher their need for information. The most common perceived advantage of IBR was the avoidance of an external prosthesis. A majority of patients were satisfied with the breast reconstruction. However, a sizeable proportion needed more information about breast reconstruction and the use of the silicone prosthesis. To avoid too high expectations more attention should be given to possible complications and the moderate cosmetic results. PMID- 10838000 TI - Psychological distress two years after diagnosis of breast cancer: frequency and prediction. AB - The present prospective study aimed at (1) investigating the frequency of high levels of psychological distress in women with early-stage breast cancer almost two years after diagnosis and (2) identifying characteristics associated with long-term distress. One hundred and seventy women participated on two occasions. Two months after surgery, patients completed questionnaires measuring psychosocial variables (e.g., stressful life-events, health complaints, sleep problems, social support, subjective distress, personality factors), demographic and biomedical variables (e.g., TNM status, type of surgery). At the second measurement, subjective distress was assessed for a second time by means of the Impact of Events Scale (IES). Almost two years after diagnosis, 16% of the women reported a high level of psychological distress as measured by the Intrusion scale (IES). Best predictors of a high level of distress were: intrusive thoughts about the disease, trait-anxiety, health complaints and problems with sleeping. No significant association was found between previous life-events, social support or biomedical variables and levels of distress. PMID- 10838001 TI - Group rehabilitation for cancer patients: satisfaction and perceived benefits. AB - Satisfaction with a group rehabilitation programme (GR) was evaluated in a heterogeneous group of cancer patients. Of the patients that were invited, 67% (N = 132) participated in the GR. The GR included eight sessions plus one booster session, each including information and/or cognitive-behavioural techniques combined with physical training and relaxation and started approximately 4 months after diagnosis. Men and women participated to the same extent. A mailed questionnaire was used to assess patients' satisfaction, perceived benefits and level of difficulty of the GR components. The majority of patients stated that the number of sessions and timing of the GR was adequate. The usefulness of the GR components were rated in the following descending order: relaxation, physical training, encountering others in the same situation, breathing exercises, information and cognitive-behavioural skills. Patients were more satisfied with diagnosis-specific group meetings than with those including several diagnoses. Assessment of patient satisfaction seems appropriate to elucidate patient priorities. PMID- 10838002 TI - Structuring psychosocial care in pediatric oncology. AB - The requirements for high quality psychosocial care for children with cancer and their families are steadily increasing. As an additional supplement to medical and nursing care, psychosocial work has to consider both the requirements of the medical treatment and the associated stress factors. It is essential to structure the different possibilities of intervention in a way which guarantees the practice of psychosocial care within the ongoing medical care. This requires a standardized psychosocial care manual. In July 1994, a 'Manual for Psychological Care in Pediatric Oncology' was put into practice and has been continually improved in the Department of Pediatric Hematology/Oncology of the University of Bonn. It contains special indications for different standards of psychosocial care, is oriented according to the phases and situations of medical treatment, and consists of a health education program as well as special care measurements. The latter are related to both the stressors primarily caused by the requirements of familial adherence to medical and nursing care and the stressors primarily caused by factors of the individual or the familial life circumstances. The theoretical basis of this manual and concrete information for its use are described in this paper. PMID- 10838003 TI - Attitude and knowledge of attention deficit hyperactivity disorder and learning disability among high school teachers. AB - The aims of the study were to investigate teachers' knowledge and attitudes towards attention deficit hyperactivity disorder (ADHD) and learning disabilities (LD). Forty-six high school teachers were interviewed in this regard. The 46 teachers were divided into two groups: 25 teachers taught at an academic school (School 1); and 21 teachers taught at special education school (School 2) and dealt with ADHD/LD cases regularly. General knowledge about ADHD (71%) and about LD (74%) was relatively low among both groups. Thirteen percent of all teachers considered LD to be the result of parental attitudes, namely 'spoiling' the children. The score for attitude and understanding of ADHD children was relatively low (72.5%) for both groups, whereas Group B teachers scored higher regarding LD cases. Almost 40% considered that ADHD children should be rebuked and/or punished in a manner similar to non-ADHD kids. Regarding long-term outcome, 45.7% of the teachers expected ADHD children to experience multiple difficulties in family life during adulthood. In relation to LD cases, the overall scoring for positive attitude was 75%. However, this score was higher for Group B teachers. Three-quarters of the teachers favored increasing peer awareness and comprehension as to the problems LD kids encounter at school. Ninety-five percent believed LD patients should enjoy a more lenient school education. There was no correlation between teachers, knowledge of ADHD and LD and their attitude. The main sources for this knowledge were: specialized textbooks, continuous education, TV shows, journals and newspapers, and medical personnel. PMID- 10838004 TI - Structured parent education in the management of childhood atopic dermatitis: the Berlin model. AB - Childhood atopic dermatitis (AD) is a common disease with the prevalence rates increasing. Its chronic course with frequent relapses puts a special burden on both children and their parents. To maximise positive long-term outcome in the management of AD it is important to support parents in dealing with the chronic condition of their child in addition to treating symptoms. In the present article, we describe in detail the goals, structure, and content of the Berlin education program for parents of children with AD. The program aims to contribute towards a comprehensive, family-oriented management of childhood AD. Its objective is to improve parent's self-management skills with regard to their child's disease and to positively impact the course of the disease as well as the family's quality of life. Medical, nutritional and psychological issues are covered in six group sessions which are conducted by a multiprofessional team of paediatricians, psychologists and dieticians. Preliminary data show that the program has a desirable effect on aspects of quality of life and coping. PMID- 10838005 TI - Change counselling in diabetes: the development of a training programme for the diabetes team. AB - Skills in counselling for behaviour change may help staff working in diabetes care to facilitate self management in people with diabetes. A feasibility study was conducted to define the essential competencies (attitudes, knowledge and skills), training methods, and assess whether this approach was practicable in a diabetes resource centre. Semi structured interviews with staff were routinely conducted throughout the duration of the study. The findings suggest that the stages of change model, motivational interviewing and behavioural techniques are relevant to work in this area. Acquiring the competencies was harder to achieve than anticipated, though most were evident after one years training. The most valued training methods were individual supervision and video examples. However, the competencies were difficult to apply in the clinical setting given time constraints, the strength of existing staff routines and the patients' readiness to change. The lessons learned and suggestions for future work are presented. PMID- 10838006 TI - Imaging outcomes from the National Library of Medicine's Visible Human Project. PMID- 10838007 TI - Exploring the Visible Human using the VOXEL-MAN framework. AB - In principle the Visible Human data sets are an ideal basis for building electronic atlases. While it is easy to construct such atlases by just offering the possibility of browsing through the 2D slices, constructing realistic 3D models is a huge project. As one rather easy way to establish 3D use, we have registered the Visible Human data to the already existing 3D atlas VOXEL MAN/brain. This procedure enables one to lookup anatomical detail in an atlas based on radiological images. Concerning the segmentation problem, which is the prerequisite for a real 3D atlas, we have developed an interactive classification method that delivers realistic perspective views of the Visible Human. As these volume based methods require high-end workstations, we finally have developed a multimedia program that runs on standard PCs and uses Quicktime VR movies. PMID- 10838008 TI - Virtual endoscopy: development and evaluation using the Visible Human datasets. AB - Virtual endoscopy (VE) is a new method of diagnosis using computer processing of 3D image datasets (such as CT or MRI scans) to provide simulated visualizations of patient specific organs similar or equivalent to those produced by standard endoscopic procedures. Conventional endoscopy is invasive and often uncomfortable for patients. It sometimes has serious side effects such as perforation, infection and hemorrhage. VE visualization avoids these risks and can minimize difficulties and decrease morbidity when used before actual endoscopic procedures. In addition, there are many body regions not compatible with real endoscopy that can be explored with VE. Eventually, VE may replace many forms of real endoscopy. There remains a critical need to refine and validate VE visualizations for routine clinical use. We have used the Visible Human Dataset from the National Library of Medicine to develop and test these procedures and to evaluate their use in a variety of clinical applications. We have developed specific clinical protocols to compare virtual endoscopy with real endoscopy. We have developed informative and dynamic on-screen navigation guides to help the surgeon or physician interactively determine body orientation and precise anatomical localization while performing the VE procedures. Additionally, the adjunctive value of full 3D imaging (e.g. looking "outside" of the normal field of view) during the VE exam is being evaluated. Quantitative analyses of local geometric and densitometric properties obtained from the virtual procedures ("virtual biopsy") are being developed and compared with other direct measures. Preliminary results suggest that these virtual procedures can provide accurate, reproducible and clinically useful visualizations and measurements. These studies will help drive improvements in and lend credibility to VE procedures and simulations as routine clinical tools. VE holds significant promise for optimizing endoscopic diagnostic procedures, minimizing patient risk and morbidity, and reducing health care costs. PMID- 10838009 TI - "True" color surface anatomy: mapping the Visible Human to patient-specific CT data. AB - The mapping of "true" color and texture information into traditional medical imaging modality data can add considerable information and aid in diagnostics. One of the goals of this work has been to create CT color lookup tables for all visually well-defined structures in the Visible Human male cryosection data set which then can be used to color patient-specific CT data. The primary goal has been to develop a method for stripping textures from a volumetric data set for polygonal models and non-uniform rational B-spline (NURBS) models generated from a volumetric data set. It is believed that these methods can eventually be used to provide clinicians with 3D models with physiologically accurate color textures. PMID- 10838010 TI - Development of a human body model for numerical calculation of electrical fields. AB - Knowledge of the distribution of electrical fields in the human body is of importance for scientists, engineers and physicians. This paper shows one way to achieve this knowledge by numerical calculation based on macroscopic models of the human body. An anatomical model is created by preprocessing, segmentation and classification of the digital images within the Visible Man data set. Conductivity models are derived, which describe the distribution of electrical conductivity in the human body. A conductivity model is applied to solve an exemplary forward problem in electrophysiology, which consist of the calculation of the electrical field distribution arising from cardiac sources. The cardiac sources are obtained by a model of the excitation process within the heart. The calculation of electrical fields is carried out numerically by employing the finite difference method. PMID- 10838011 TI - Semi-automated color segmentation of anatomical tissue. AB - We propose a semi-automated region-based color segmentation algorithm to extract anatomical structures, including soft tissues, in the color anatomy slices of the Visible Human data. Our approach is based on repeatedly dividing an image into regions using Voronoi diagrams and classifying the regions based on experimental classification statistics. The user has the option of reclassifying regions in order to improve the final boundary. Our results indicate that the algorithm can find accurate outlines in a small number of iterations and that manual interaction can markedly improve the outline. This approach can be extended to 3D color segmentation. PMID- 10838012 TI - Radiometric homogenization of the color cryosection images from the VHP lungs for 3D segmentation of blood vessels. AB - This work deals with the problem of radiometric inhomogeneities found on the physical color images of the anatomical cryosections from the Visible Human Project (VHP) male body. Our goal is to extract very thin structures, like the blood vessel tree from the lungs. Current segmentation methods applied to VHP color images are disturbed by discontinuous, inter-slice radiometric variations; we thus devised an adaptive correction that is propagated along a series of parallel slices, taking advantage of the structural coherence between consecutive slices. No blurring is introduced, and fine details and texture are respected. Results of 3D segmentation of fine blood vessels on the corrected volume are presented. PMID- 10838013 TI - 3D shape recovery of a newborn skull using thin-plate splines. AB - The objective of this paper is to construct a mesh-model of a newborn skull for finite element analysis to study its deformation when subjected to the forces present during labour. The current state of medical imaging technology has reached a level which allows accurate visualisation and shape recovery of biological organs and body-parts. However, a sufficiently large set of medical images cannot always be obtained, often because of practical or ethical reasons, and the requirement to recover the shape of the biological object of interest has to be met by other means. Such is the case for a newborn skull. A method to recover the three-dimensional (3D) shape from (minimum) two orthogonal atlas images of the object of interest and a homologous object is described. This method is based on matching landmarks and curves on the orthogonal images of the object of interest with corresponding landmarks and curves on the homologous or 'master'-object which is fully defined in 3D space. On the basis of this set of corresponding landmarks, a thin-plate spline function can be derived to warp from the 'master'-object space to the 'slave'-object space. This method is applied to recover the 3D shape of a newborn skull. Images from orthogonal view-planes are obtained from an atlas. The homologous object is an adult skull, obtained from CT images made available by the Visible Human Project. After shape recovery, a mesh model of the newborn skull is generated. PMID- 10838014 TI - Characterization of chitosan hydrochloride-mucin interaction by means of viscosimetric and turbidimetric measurements. AB - In the present work the interaction between chitosan hydrochloride (HCS) and two different types of mucin - one obtained from bovine submaxillary glands and the other from porcine stomach - was investigated. Two hydration media were tested: distilled water and 0. 1 M HCl. Intrinsic viscosity, which provides information about polymeric chain conformation, was assessed in both media for HCS and bovine submaxillary mucin. Changes in the specific viscosity of HCS-mucin mixtures were observed as a function of the polymer:mucin weight ratio. The formation of interaction products was indicated by a minimum in the specific viscosity. Such a minimum occurred at different polymer:mucin weight ratios depending on the hydration medium and mucin type. This suggested a different stoichiometry of interaction. Turbidimetric measurements were also effected in order to evidentiate the eventual precipitation of the polymer-mucin interaction products. While in distilled water the precipitation of the interaction product did occur, in acidic medium, although a minimum in specific viscosity was observed, no precipitate was formed. The two techniques employed, viscosimetric and turbidimetric, allowed us to investigate for both mucins the influence of hydration medium on the formation of the HCS-mucin interaction products and to conclude that a slightly acid-neutral pH favours the interaction between HCS and mucins. PMID- 10838015 TI - Binding of [3H]MSX-2 (3-(3-hydroxypropyl)-7-methyl-8-(m-methoxystyryl)-1 propargylxanthine) to rat striatal membranes--a new, selective antagonist radioligand for A(2A) adenosine receptors. AB - The present study describes the preparation and binding properties of a new, potent, and selective A(2A) adenosine receptor (AR) antagonist radioligand, [3H]3 (3-hydroxypropyl)-7-methyl-8-(m-methoxystyryl)-1-propargy lxanth ine ([3H]MSX-2). [3H]MSX-2 binding to rat striatal membranes was saturable and reversible. Saturation experiments showed that [3H]MSX-2 labeled a single class of binding sites with high affinity (K(d)=8.0 nM) and limited capacity (B(max)=1.16 fmol.mg( 1) of protein). The presence of 100 microM GTP, or 10 mM magnesium chloride, respectively, had no effect on [3H]MSX-2 binding. AR agonists competed with the binding of 1 nM [3H]MSX-2 with the following order of potency: 5'-N ethylcarboxamidoadenosine (NECA)>2-[4-(carboxyethyl)phenylethylamino]-5'-N ethylcarboxami doaden osine (CGS-21680)>2-chloroadenosine (2-CADO)>N(6) cyclopentyladenosine (CPA). AR antagonists showed the following order of potency: 8-(m-bromostyryl)-3, 7-dimethyl-1-propargylxanthine (BS-DMPX)>1, 3-dipropyl-8 cyclopentylxanthine (DPCPX)>(R)-5, 6-dimethyl-7-(1-phenylethyl)-2-(4-pyridyl)-7H pyrrolo[2, 3-d]pyrimidine-4-amine (SH-128)>3,7-dimethyl-1-propargylxanthine (DMPX)>caffeine. The K(i) values for antagonists were in accordance with data from binding studies with the agonist radioligand [3H]CGS21680, while agonist affinities were 3-7-fold lower. [3H]MSX-2 is a highly selective A(2A) AR antagonist radioligand exhibiting a selectivity of at least two orders of magnitude versus all other AR subtypes. The new radioligand shows high specific radioactivity (85 Ci/mmol, 3150 GBq/mmol) and acceptable nonspecific binding at rat striatal membranes of 20-30%, at 1 nM. PMID- 10838016 TI - The development of an instrumented tamp-filling capsule machine I. Instrumentation Of a Bosch GKF 400S machine and feasibility study. AB - The instrumentation of capsule filling machines operating by the tamp-filling principle has been described only once, using strain gauges. In the present paper the instrumentation of a Bosch GKF 400S tamp-filling machine has been described using a prototype of a pneumatic tamping head equipped with a piezoelectric force transducer. The pneumatic system replaces the conventional springs situated between tamping pins and upper part of the tamping head. Via a feedback switch valve, the air pressure inside the pneumatic chamber can be regulated. This provides a potential mechanism for feedback control of capsule fill weight during continuous capsule filling. It was found that the use of the pneumatic tamping head is limited to the control of fill weight during tamping. Major adjustments of fill weight at the set-up stage of the machine should be made by alteration of the tamping pin and powder bed height settings. The principles of capsule fill weight control by continuous monitoring of tamping forces have been established, but the transfer of the system to full industrial use requires further development of the prototype by the machine manufacturer. PMID- 10838017 TI - Development of a sequential linked pharmacokinetic and pharmacodynamic simulation model for ivabradine in healthy volunteers. AB - Ivabradine is a novel bradycardic agent that has been developed for the prevention of angina. Ivabradine has an active metabolite S-18982. The aim of the study was to develop a linked pharmacokinetic-pharmacodynamic simulation model for the description of exercise-induced heart rate. The pharmacodynamic data (heart rate) were pooled from two studies and included a total of 78 healthy subjects. The data consisted of multiple dose oral administration of ivabradine. The multiple dose regimens were administered every 12 h. There were eight active dosing levels and placebo, and a no-dose run in the period before each study. The modelling was performed using the NONMEM software. Both ivabradine and S-18982 possess bradycardic activity, although the extent of the activity of both could not be determined from the data available. A multiple ligand pharmacodynamic model provided the best fit to the data. The model was assessed in terms of its posterior predictive performance and was able to describe the original data adequately when used for simulation purposes. PMID- 10838018 TI - A pharmacokinetic simulation model for ivabradine in healthy volunteers. AB - Ivabradine is a novel bradycardic agent that has been developed for the prevention of angina. Ivabradine has an active metabolite S-18982. The aim of this study is to develop a pharmacokinetic simulation model. Pharmacokinetic data from two studies were pooled and included data from a total of 66 healthy male volunteers. The data were collected following single dose intravenous and multiple dose oral administration of ivabradine. The multiple dose regimens were administered every 12 h and there were seven active dosing levels. The modelling was performed using the NONMEM software. The model was assessed in terms of its ability to describe the original data set used in its construction and also data arising from a different clinical pharmacology study involving 12 additional subjects. The pharmacokinetics of ivabradine and S-18982 were best described by two linked two compartment intravenous bolus and first-order input, with first pass loss, and first-order output model. When the model was used for simulation it produced an adequate description of both the original data and data arising from a different clinical pharmacology study. PMID- 10838019 TI - Theoretical calculation and prediction of P-glycoprotein-interacting drugs using MolSurf parametrization and PLS statistics. AB - A method for the modelling and prediction of P-glycoprotein-associated ATPase activity using theoretically computed molecular descriptors and multivariate statistics has been investigated using 22 diverse drug-like compounds. The program MolSurf was used to compute theoretical molecular descriptors related to physicochemical properties such as lipophilicity, polarity, polarizability and hydrogen bonding. The multivariate partial least squares projections to latent structures (PLS) method was used to delineate the relationship between the P glycoprotein-associated ATPase activity and the theoretically computed molecular descriptors. The PLS analysis of the entire data set, with the exclusion of tamoxifen, resulted in one significant PLS component according to cross validation with R(2)=0.718, Q(2)=0. 695, S.D.=0.475, F=48.37, RMSE(tr)=0.452, p<0.001. Properties associated with the size of the molecular surface, polarizability and hydrogen bonding had the largest impact on the P-glycoprotein associated ATPase activity. All these properties should be high to promote high ATPase activity. PMID- 10838020 TI - Preparation and characterization of solid lipid nanospheres containing paclitaxel. AB - The study describes the development of stealth and non-stealth solid lipid nanospheres (SLNs) as colloidal carriers for paclitaxel, a drug with very low solubility. SLNs are constituted mainly of bioacceptable and biodegradable lipids, such as tripalmitin and phosphatidylcholine, and can incorporate amounts of paclitaxel up to 2.8%. Stealth and non-stealth loaded SLNs are in the nanometer size range and can be sterilized and freeze-dried. Thermal analysis (differential scanning calorimetry) showed that paclitaxel is not able to crystallize in the SLNs. Release of paclitaxel from SLNs is very low. Non-stealth and stealth SLNs are stable over time without precipitation of paclitaxel and can be proposed for its parenteral administration. PMID- 10838021 TI - Physicochemical characterization of solid dispersions of the antiviral agent UC 781 with polyethylene glycol 6000 and Gelucire 44/14. AB - The purpose of this study was to prepare and characterize solid dispersions of the antiviral thiocarboxanilide UC-781 with PEG 6000 and Gelucire 44/14 with the intention of improving its dissolution properties. The solid dispersions were prepared by the fusion method. Evaluation of the properties of the dispersions was performed using dissolution studies, differential scanning calorimetry, Fourier-transform infrared spectroscopy and X-ray powder diffraction. To investigate the possible formation of solid solutions of the drug in the carriers, the lattice spacings [d] of PEG 6000 and Gelucire 44/14 were determined in different concentrations of UC-781. The results obtained showed that the rate of dissolution of UC-781 was considerably improved when formulated in solid dispersions with PEG 6000 and Gelucire 44/14 as compared to pure UC-781. From the phase diagrams of PEG 6000 and Gelucire 44/14 it could be noted that up to approximately 25% w/w of the drug was dissolved in the liquid phase in the case of PEG 6000 and Gelucire 44/14. The data from the X-ray diffraction showed that the drug was still detectable in the solid state below a concentration of 5% w/w in the presence of PEG 6000 and Gelucire 44/14, while no significant changes in the lattice spacings of PEG 6000 or Gelucire 44/14 were observed. Therefore, the possibility of UC-781 to form solid solutions with the carriers under investigation was ruled out. The results from infrared spectroscopy together with those from X-ray diffraction and differential scanning calorimetry showed the absence of well-defined drug-polymer interactions. PMID- 10838022 TI - Haliotis tuberculata hemocyanin (HtH): analysis of oligomeric stability of HtH1 and HtH2, and comparison with keyhole limpet hemocyanin KLH1 and KLH2. AB - The multimeric/higher oligomeric states of the two isoforms of Haliotis tuberculata hemocyanin (HtH1 and HtH2) have been assessed by transmission electron microscopy (TEM) of negatively stained specimens, for comparison with previously published structural data from keyhole limpet hemocyanin (KLH1 and KLH2) [see Harris, J.R., Gebauer, W., Guderian, F.U., Markl, J., 1997a. Keyhole limpet hemocyanin (KLH), I: Reassociation from Immucothel followed by separation of KLH1 and KLH2. Micron, 28, 31-41; Harris, J.R., Gebauer, W., Sohngen, S.M., Nermut, M.V., Markl, J., 1997b. Keyhole limpet hemocyanin (KLH). II: Characteristic reassociation properties of purified KLH1 and KLH2. Micron, 28, 43 56; Harris, J.R., Gebauer, W., Adrian, M., Markl, J., 1998. Keyhole limpet hemocyanin (KLH): Slow in vitro reassociation of KLH1 and KLH2 from Immucothel. Micron, 29, 329-339]. In purified samples of both HtH isoforms, the hollow cylindrical ca 8MDa didecamer predominates together with a small number of decamers, but tri- and longer multidecamers are detectable only in the HtH2. The stability of the two HtH isoforms under varying ionic conditions have been monitored, thereby enabling conditions for the production of stable decamers to be established. The ability of these decamers to reform multimers in the presence of 10 and 100mM concentrations of calcium and magnesium ions in Tris-HCl buffer (pH 7.4), and also of individual HtH1 and HtH2 subunits (produced by pH 9.6 dissociation in glycine-NaOH buffer), to reassociate in the presence of calcium and magnesium ions, has been assessed. For the HtH1 decamers, the predominant multimeric product is the didecamer at 10 and 100mM calcium and magnesium concentrations, whereas for the HtH2 decamers, large numbers of multidecamers are produced, with the reaction proceeding more completely at the higher calcium and magnesium concentration. With the HtH1 subunit, reassociation in the presence of 10 and 100mM calcium and magnesium ions yielded an almost 100% conversion into didecamers, whereas the HtH2 subunit produced a mixture containing large numbers of short multidecamers and relatively few didecamers, together with a considerable number of smaller diameter helical/tubular polymers. The association properties of the HtH1 and HtH2 decamers, and the subunit reassociation, firmly indicate the integrity and structural competency of the protein under the experimental conditions used. Data on the association of KLH2 decamers is also presented, which together with previously published data on the association KLH1 decamers and the reassociation of KLH1 and KLH2 subunits, enables a detailed comparison of the two hemocyanin isoforms from both molluscan species to be made. Biochemical manipulation of the oligomer states and the subunit reassociation of molluscan hemocyanins can usefully be assessed by the study of negatively stained TEM specimens. PMID- 10838024 TI - The spalling of overgrowths during experimental freeze-thaw of a quartz sandstone as a mechanism of quartz silt production AB - The experimental frost weathering of a quartz sandstone is being carried out as part of a series of laboratory simulations to investigate geomorphic mechanisms capable of producing loess-sized quartz silt. The preliminary results are reported in this paper. Frost weathering simulations involved two different temperature regimes: (a) -5 degrees C-+15 degrees C, and (b) -12 degrees C-+15 degrees C. Debris production was observed for both temperature regimes with the blocks subjected to the more intense freezing regime experiencing the most disintegration. Scanning electron microscopic (SEM) examination and particle size analysis revealed that the debris produced by both these experiments included a significant silt sized fraction. Further SEM examinations of this debris demonstrated the presence of silt sized fragments of quartz overgrowths among the debris. It is suggested that overgrowth detachment occurs during frost weathering by either the penetration into and subsequent freezing of water in the gaps between overgrowths and host grains, or pressures applied to the outer surfaces of overgrowths during frost cycles causes them to collapse onto the underlying grain. PMID- 10838023 TI - A morphological study of the effect of chlorambucil during the S and G2 phases of the cell cycle of synchronized HEp-2 cancer cell populations using computerized morphometry. AB - Chlorambucil, a bisalkylating agent, used extensively in the treatment of autoimmune and neoplastic diseases, is known to affect DNA synthesis. However recent studies have revealed that it also affects the synthesis of other nuclear protein constituents, especially histones. Since histones play a major role in both the structural and functional integrity of chromatin, we have analyzed the morphological effects of this agent, using low dose conditions and synchronized populations of HEp-2 cancer cells in the S and G2 phases of the cell cycle. Analyses at the light and electron microscopy levels were undertaken using synchronous image analysis techniques. Computerized morphometry was used so as to evaluate various nuclear and cytological morphological parameters. It was found that chlorambucil affects the organization of chromatin, as well as other cellular parameters in a manner characteristic of decreased tumor aggressiveness. A finding of significance in this study was that chlorambucil exerted its influence on all these morphological parameters only when treatment was initiated at the beginning of the S phase and not during the second half of the S phase or the G2 phase. PMID- 10838025 TI - Antigen retrieval on formaldehyde-fixed paraffin sections: its potential drawbacks and optimization for double immunostaining. AB - It is well known that the major artifact induced by formaldehyde fixation is the masking of tissue antigens due to cross-linking of protein amino acid residues. Recently many antigen retrieval techniques have been devised to unmask the hidden antigen epitopes and recover immunoreactivity. In this study, some practical problems of two common unmasking techniques, i.e. heat-induced epitope retrieval and enzyme digestion have been reviewed in immunostaining of proliferating cell nuclear antigen (PCNA) on formaldehyde-fixed paraffin-embedded sections. As the heating conditions became more severe, false-positive staining and/or nonspecific background staining occurred. Based on the principle of protein inactivation/denaturation and the possible mechanisms of antigen retrieval, it has been suggested that the antigen retrieval itself can also denature proteins in tissues, just as many other protein inactivation processes. Thus, the total magnitude of protein conformational change caused by the overall unmasking procedure is in practice crucial. To prove this hypothesis and to overcome such undesirable drawbacks after antigen retrieval, a new combination technique of a mild heating condition (microwaved at 80 degrees C for 15-20min) and pepsin digestion was devised. This technique led to a strong specific immunoreactivity of PCNA, without any undesirable false positive or background staining. The procedure was also adapted for double immunostaining of PCNA together with alpha actin, bromodeoxyuridine, keratin, type IV collagen and vimentin. PMID- 10838026 TI - Detection of Au precipitates in a Mg-based alloy using electronically simulated hollow cone illumination AB - Images from electronically simulated hollow cone illumination (or rotating dark field) conditions, obtained under plane wave (or weak) diffraction conditions, are generally assumed to approximate to the compositionally weighted sum of atomic number squared for sufficiently large momentum transfers. However, even for large momentum transfer encountered with a semi-angle of 5 degrees and 300keV electrons, appropriate numerical integration over condenser and objective aperture configurations indicates that some thermal scattering component is still present. A Mg-Al alloy with minor additions of Zn and Mn, and to which 0.1at.% Au has been added, is shown to provide a good system for the detection of high Z (atomic number) precipitates within a low Z matrix and on which semi-quantitative calculations may be based. Correlation of absolute rather than relative intensities from small precipitates (3-10nm diameter) with calculations based on an Einstein model for (incoherent) thermal diffuse scattering show that the small precipitates consist predominantly of Au, a conclusion subsequently supported by EDX analysis and electron diffraction measurements. It is also demonstrated that this incoherent contrast mechanism is ideal for stereographic imaging in the TEM. PMID- 10838027 TI - Atomic force microscopy analysis of wool fibre surfaces in air and under water AB - Wool fibre surfaces have been treated by solvent cleaning which leaves the native covalently bound surface lipid layer intact, and by alcoholic alkali which removes the lipid layer. The resultant surfaces have been analysed by atomic force microscopy (AFM), with particular emphasis on force-distance (F-d) methods. Methodologies were developed for investigation in situ in water of both the surface topography and the characteristics of the lipid layer. Longitudinal surface texturing was resolved in images of wool fibre surfaces in air; the texturing remained prominent after exposure to water. High resolution F-d curves revealed features associated with the lipid layer. A simple formalism was used to show that the layer had a thickness of a few nm, and an effective stiffness of some 0. 12+/-0.01N/m. Strong adhesive interactions, equivalent to a pressure of 0.1MPa, acted on the tip at the tip-to-substrate interface. The methodology and formalism are likely to be relevant in the broad field of thin-film analysis and for fibre technology. PMID- 10838028 TI - Cryo-scanning electron microscopic study on freezing behavior of xylem ray parenchyma cells in hardwood species AB - Differential thermal analysis (DTA) has indicated that xylem ray parenchyma cells (XRPCs) of hardwood species adapt to freezing of apoplastic water either by deep supercooling or by extracellular freezing, depending upon the species. DTA studies indicated that moderately cold hardy hardwood species exhibiting deep supercooling in the XRPCs were limited in latitudinal distribution within the -40 degrees C isotherm, while very hardy hardwood species exhibiting extracellular freezing could distribute in colder areas beyond the -40 degrees C isotherm. Predictions based on the results of DTA, however, indicate that XRPCs exhibiting extracellular freezing may appear not only in very hardy woody species native to cold areas beyond the -40 degrees C isotherm but also in less hardy hardwood species native to tropical and subtropical zones as well as in a small number of moderately hardy hardwood species native to warm temperate zones. Cryo-scanning electron microscopic (cryo-SEM) studies on the freezing behavior of XRPCs have revealed some errors in DTA. These errors are originated mainly due to the overlap between exotherms produced by freezing of water in apoplastic spaces (high temperature exotherms, HTEs) and exotherms produced by freezing of intracellular water of XRPCs by breakdown of deep supercooling (low temperature exotherms, LTEs), as well as to the shortage of LTEs produced by intracellular freezing of XRPCs. In addition, DTA results are significantly affected by cooling rates employed. Further, cryo-SEM observations, which revealed the true freezing behavior of XRPCs, changed the previous knowledge of freezing behavior of XRPCs that had been obtained by freeze-substitution and transmission electron microscopic studies. Cryo-SEM results, in association with results obtained from DTA that were reconfirmed or changed by observation using a cryo-SEM, revealed a clear tendency of the freezing behavior of XRPCs in hardwood species to change with changes in the temperature in the growing conditions, including both latitudinal and seasonal temperature changes. In latitudinal temperature changes, XRPCs in less hardy hardwood species native to tropical and subtropical zones exhibited deep supercooling to -10 degrees C, XRPCs in moderately hardy hardwood species native to temperate zones exhibited a gradual increase in the supercooling ability to -40 degrees C from warm toward cool temperate zones, and XRPCs in very hardy hardwood species native to boreal forests exhibited extracellular freezing via an intermediate form of freezing behavior between deep supercooling and extracellular freezing. In seasonal temperature changes, XRPCs in hardwood species native to temperate zones changed their supercooling ability from a relatively low degree in summer to a high degree in winter. XRPCs in hardwood species native to boreal forests changed their freezing behavior from deep supercooling to -10 degrees C in summer to extracellular freezing in winter. These results indicate that the freezing behavior of XRPCs in hardwood species tends to shift gradually from supercooling of -10 degrees C, to a gradual increase in the deep supercooling ability to -40 degrees C or less, and finally to extracellular freezing as a result of cold acclimation in response to both latitudinal and seasonal temperature changes. It is thought that these temperature-dependent changes in the freezing behavior of XRPCs in hardwood species are mainly controlled by changes in cell wall properties, although no distinct changes were detected by electron microscopic observations in cell wall organization between hardwood species or between seasons. Evidence of temperature dependent changes in the freezing behavior of XRPCs in hardwood species provided by the results of studies using a cryo-SEM has indicated the need for further investigation to clarify cold acclimation-induced cell wall changes at the sub electron microscopic level in order to understand the mechanisms of freezing adaptation. PMID- 10838029 TI - Reconstruction principles of icosahedral virus structure determination using electron cryomicroscopy. AB - Electron cryomicroscopy is a useful tool for studying the three-dimensional structure of icosahedral viruses. This review is intended to provide beginners with an understanding of icosahedral virus structure determination focusing on the data processing aspects. We begin with an overview of the entire structure determination process and a brief summary of the sample preparation and imaging aspects. Next, we provide detailed descriptions of each data processing step leading to three-dimensional reconstruction, including application of image corrections, resolution assessment, and structure visualization. To aid in understanding this reconstruction process we provide a variety of illustrative examples. Last, we summarize future prospects for icosahedral virus structural studies. PMID- 10838030 TI - Towards reconciliation of structure with function in plasmodesmata-who is the gatekeeper? AB - Whilst the structure of higher plant plasmodesmata was first described by Robards (1963. Desmotubule-a plasmodesmatal substructure. Nature 218, 784), and despite many subsequent intensive investigations, there is still much that remains unclear relating to their ultrastructure and functioning in higher plants. We have examined chemically fixed plant material, and suggest that the conformational changes seen in plasmodesmatal substructure, particularly the deposition of electron-dense extra-plasmodesmal material, is linked to either manipulation of the hormonal balance (as in Avocado fruit), or of osmotic potential in leaf blade material. These changes result in the deposition of beta 1,3-glucan (callose) at the neck region of these plasmodesmata. This electron dense material is deposited at the neck region of plasmodesmata, and forms a collar-like structure. The formation of a collar is shown to be coupled with loss of lucence within the cytoplasmic sleeve. The formation of a collar at the plasmodesmatal orifice thus results in encapsulation and closure of the plasmodesmatal orifice. Closure of the orifice coincides with a loss of electron lucence and a lack of resolution of the desmotubule. These ultrastructural changes are potentially significant and could contribute to, result in, or assist in the down-regulation of cell to cell trafficking via plasmodesmata. PMID- 10838031 TI - Analysis of two major anti-M2 antibodies (anti-PDC-E2/anti-BCOADC-E2) in primary biliary cirrhosis: relationship to titers of immunofluorescent anti-mitochondrial antibody. AB - To analyze anti-M2 components in primary biliary cirrhosis (PBC) we measured two major anti-M2 antibodies (anti-PDC-E2 and anti-BCOADC-E2) by immunoblotting and ELISA, and compared the results between 38 immunofluorescent anti-mitochondrial antibody (AMA)-negative PBC patients (group A) and 39 strongly AMA-positive PBC patients (group B) with titers of 1:640. Using bovine heart mitochondrial fraction as antigen, the immunoblot positivity rate of anti-PDC-E2 in group B was significantly higher than that in group A, whereas the positivity rate of anti BCOADC-E2 was not significantly different between the two groups. This result was similar to that obtained by ELISA using recombinant fusion proteins. In group A there was a significant inverse correlation between ELISA optical density values of anti-PDC-E2 and of anti-BCOADC-E2, but in group B there was no correlation between the two values. Only three patients from group A and 21 from group B were positive for both antibodies. Taken together these results appear to indicate that the detection of anti-BCOADC-E2 is critical for the accurate serological diagnosis of AMA-negative PBC patients. The detection of anti-BCOADC-E2 may also help to distinguish between AMA-negative PBC and autoimmune cholangitis patients. PMID- 10838032 TI - Serum alpha-glutathione S-transferase: a new marker of hepatocellular damage associated with hepatectomy. AB - Serum concentrations of alpha-glutathione S-transferase (alphaGST) were determined before and after hepatectomy to examine the clinical usefulness of alphaGST as a marker of hepatocellular damage compared with the conventional liver function tests. Prior to hepatectomy, serum alphaGST concentrations correlated significantly with serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT). In patients who had a good postoperative course, serum alphaGST concentrations rose significantly to a peak immediately after surgery, followed by a rapid fall to the normal range within 1 week, yielding a monophasic pattern. Serum alphaGST concentrations reached a peak earlier than other parameters of liver function, and peak serum alphaGST concentrations correlated with peak serum concentrations of AST and ALT. The mean decrease rate of serum alphaGST concentration from peak values was significantly more rapid than that of serum AST and ALT, indicating an early return of alphaGST concentrations to the normal range. These findings suggest that serum alphaGST may be a more sensitive marker of hepatocellular damage than transaminases and may therefore be useful for rapid monitoring of the extent and persistence of liver injury after hepatectomy. PMID- 10838033 TI - Detection of HBV RNA in peripheral blood mononuclear cells in patients with and without HBsAg by reverse transcription polymerase chain reaction. AB - We examined RNA of hepatitis B virus (HBV) in peripheral blood mononuclear cells (PBMCs) by the reverse transcription polymerase chain reaction (RT PCR) in 61 patients associated with HBV infection, in order to analyze the relationship between the transcriptional activity of HBV in PBMCs and the clinical characteristics. The presence of HBV RNA in PBMCs was detected in 19/51(37.1%) patients with HBsAg positive and in 1/10 (10.0%) patient with HBsAg negative patients. Six healthy controls were all negative. The frequency of HBV RNA positivity was detected in patients with high ALT level (P<0.05), serum HBeAg positivity (P<0.01) and serum HBV DNA level>/=0.7 Meq/ml (P<0.05). Moreover, HBV RNA in PBMCs was detected in one patient followed up for 2 years after HBsAg disappearance in serum, who had not HBV DNA but anti-HBc IgG, in serum. These results suggested that the transcription of HBV in PBMCs, was frequently detected in the patients with higher replication of the virus, but HBV RNA in PBMCs might be detected in a few patients who had no evidence of HBV replication serologically. PMID- 10838034 TI - Irsogladine upregulates expressions of connexin32 and connexin26 in the rat liver. AB - A gap junction is the channel for cell-to-cell communication and plays an important role in the maintenance of tissue homeostasis, control of cell growth and differentiation, and prevention of experimental hepatocarcino-genesis. Irsogladine, an antiulcer drug, augments gap junctional intercellular communication in gastric mucosa, but the effect of irsogladine on the liver remains uncertain. In this study the effects of irsogladine on the liver were investigated from the viewpoints of gap junctional protein connexin (Cx)32 and Cx26 in rats. Twelve rats were divided into a control group (n=6) and the irsogladine group (n=6) in which irsogladine (20 mg/kg per day) was administered orally for 3 days before sample collection, and the two groups were compared in regard to liver enzymes (serum aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH)), serum- and tissue calcium concentrations, immunohistochemical expressions of Cx32 and Cx26, and RT PCR analysis. In immunohistochemistry, analyzed using an image processor for analytical pathology (IPAP), the number of Cx32-positive spots was higher and the area of Cx26-positive spots were larger in the irsogladine group than those in the control group (P=0.036 and P=0.00032, respectively). In RT-PCR analysis, the mRNA of Cx32 or Cx26 in the irsogladine group showed a tendency to be higher than in the control group, but not significantly (Cx32, P=0.70; Cx26, P=0.07). Another 30 rats were used for measurements of cyclic-adenosine monophosphate (c-AMP) of the liver. c-AMP concentration was increased 1 h after the administration of irsogladine, which partially explained how the Cxs were upregulated. These findings may suggest that irsogladine upregulates Cx32 and Cx26 expressions in the liver of rats. PMID- 10838035 TI - Investigation of TTV by in situ hybridization in patients with chronic hepatitis. AB - To clarify whether TT virus (TTV) was present in liver tissues, 12 liver tissue samples from patients with chronic hepatitis positive for TTV in their serum and 11 samples from serum-negative patients were obtained by needle biopsies and investigated using in situ hybridization. Positive staining was observed in nine (75%) of 12 cases positive for TTV (serum-positive group) and three (27.3%) of 11 cases negative for TTV (serum-negative group) (P=0.061). Three kinds of staining patterns were observed: nuclear, cytoplasmic and both. In 58.3% (7/12) of the patients positive for TTV staining, the stained areas were found in both the nucleus and cytoplasm. Only cytoplasmic staining was observed in three cases from the serum-positive group. Only nuclear staining was observed in two cases from the serum-negative group. No significant differences were found in the clinical background between the in situ hybridization-positive and -negative groups, and between the serum-positive and -negative groups. The present study shows that TTV exists in the liver tissue, especially in hepatocytes, of chronic hepatitis patients and that the localization of TTV in the cell is different from case to case, although why this is so remains to be clarified. PMID- 10838036 TI - Combined phlebotomy and ursodeoxycholic acid treatment in the patients with chronic hepatitis C. AB - As an option to interferon, either iron removal by phlebotomy or ursodeoxycholic acid administration has been recommended for patients with chronic hepatitis C. Some patients, however, show only partial responses to such monotherapy. In the present study, we investigated the effects of a combination of phlebotomy and ursodeoxycholic acid in the patients who did not show normalization of serum alanine aminotransferase levels by either phlebotomy or ursodeoxycholic acid monotherapy. The combination of these therapies in any order additively improved the biochemical parameter to the upper normal range. There were no statistically significant differences between the results of the two combination treatments. The combination treatment was also effective in decreasing serum alpha fetoprotein levels. PMID- 10838037 TI - An assessment of the clinical utility of serum ALT and AST in chronic hepatitis C. AB - Background: Alanine aminotransferase (ALT) is frequently used as the sole biochemical marker for chronic hepatitis C (CHC), however, its value may be normal in cases with active disease. Recently, aspartate aminotransferase (AST) has been suggested as a useful predictor of liver pathology and conflicting results were obtained by using AST/ALT ratio to predict cirrhosis. Aims: To evaluate clinical utility of serum ALT and AST in CHC. Methods: The charts of 133 patients with CHC, whose ALT and AST were simultaneously tested from 1994 to 1996, were reviewed. ALT and AST were analyzed for both the entire cohort of patients and subgroups stratified for histopathology, age, gender, alcohol consumption, and risk factors of transmission. In 53 patients, the AST/ALT ratio was evaluated during interferon treatment. Results: The elevation of ALT significantly correlated with that of AST (r=0.79). The AST/ALT ratio increased with liver histological progression. The ratio >/=1 was predominantly in cirrhotic patients. During treatment the ratio increased. The AST remained elevated in eight of the 33 patients in whom the ALT had returned to normal during and after treatment. Conclusions: Both ALT and AST are useful markers for CHC. However, the AST may elevate alone, suggesting that measuring AST may be useful when the ALT is consistently normal. The AST/ALT ratio varies in different patients but increases with histological progression of fibrosis. An AST/ALT ratio >/=1 is highly suggestive of the presence of cirrhosis. PMID- 10838038 TI - Inhibition of tumor growth with doxorubicin in a new orthotopically implanted human hepatocellular carcinoma model. AB - A number of human xenograft orthotopic models of hepatocellular carcinoma (HCC) have been previously established by growing histologically-intact patient specimens in nude mice. However, the availability of HBV and HCV negative human hepatocellular carcinoma specimens is scarce and the pattern of tumor growth in nude mice varies depending on the tumor type. In the present study, we have established a reproducible xenograft orthotopic model using a human HCC cell line designated HuT7-3 that was derived from two rounds of subcloning of the parental Huh-7 cell line. The tumor growth rate of the HuT7-3 cell line, grown at a primary subcutaneous site, was markedly higher than that of the Huh-7 parental cell line or the human hepatoblastoma Hep-G2 cell line. Furthermore, we have shown that doxorubicin, when administered intravenously, is efficient in inhibiting the development of subcutaneous tumor but leads to the regression of the orthotopic human HCC. Consequently, this novel HCC xenograft orthotopic model can be used for the evaluation of antitumor drugs. PMID- 10838039 TI - On what makes the gamma subunit spin during ATP hydrolysis by F(1). PMID- 10838040 TI - Analysis of the nucleotide binding sites of mitochondrial ATP synthase provides evidence for a two-site catalytic mechanism. PMID- 10838041 TI - Catalytic site forms and controls in ATP synthase catalysis. AB - A suggested minimal scheme for substrate binding by and interconversion of three forms of the catalytic sites of the ATP synthase is presented. Each binding change, that drives simultaneous interchange of the three catalytic site forms, requires a 120 degrees rotation of the gamma with respect to the beta subunits. The binding of substrate(s) at two catalytic sites is regarded as sufficing for near maximal catalytic rates to be attained. Although three sites do not need to be filled for rapid catalysis, during rapid bisite catalysis some enzyme may be transiently present with three sites filled. Forms with preferential binding for ADP and P(i) or for ATP are considered to arise from the transition state and participate in other steps of the catalysis. Intermediate forms and steps that may be involved are evaluated. Experimental evidence for energy-dependent steps and for control of coupling to proton translocation and transition state forms are reviewed. Impact of relevant past data on present understanding of catalytic events is considered. In synthesis a key step is suggested in which proton translocation begins to deform an open site so as to increase the affinity for ADP and P(i), that then bind and pass through the transition state, and yield tightly bound ATP in one binding change. ADP binding appears to be a key parameter controlling rotation during synthesis. In hydrolysis ATP binding to a loose site likely precedes any proton translocation, with proton movement occurring as the tight site form develops. Aspects needing further study are noted. Characteristics of the related MgADP inhibition of the F(1) ATPases that have undermined many observations are summarized, and relations of three-site filling to catalysis are assessed. PMID- 10838042 TI - The epsilon subunit of bacterial and chloroplast F(1)F(0) ATPases. Structure, arrangement, and role of the epsilon subunit in energy coupling within the complex. AB - Recent studies show that the epsilon subunit of bacterial and chloroplast F(1)F(0) ATPases is a component of the central stalk that links the F(1) and F(0) parts. This subunit interacts with alpha, beta and gamma subunits of F(1) and the c subunit ring of F(0). Along with the gamma subunit, epsilon is a part of the rotor that couples events at the three catalytic sites sequentially with proton translocation through the F(0) part. Structural data on the epsilon subunit when separated from the complex and in situ are reviewed, and the functioning of this polypeptide in coupling within the ATP synthase is considered. PMID- 10838043 TI - The rotary binding change mechanism of ATP synthases. AB - The F(0)F(1) ATP synthase functions as a rotary motor where subunit rotation driven by a current of protons flowing through F(0) drives the binding changes in F(1) that are required for net ATP synthesis. Recent work that has led to the identification of components of the rotor and stator is reviewed. In addition, a model is proposed to describe the transmission of energy from four proton transport steps to the synthesis of one ATP. Finally, some of the requirements for efficient energy coupling by a rotary binding change mechanism are considered. PMID- 10838044 TI - Synthase (H(+) ATPase): coupling between catalysis, mechanical work, and proton translocation. AB - Coupling with electrochemical proton gradient, ATP synthase (F(0)F(1)) synthesizes ATP from ADP and phosphate. Mutational studies on high-resolution structure have been useful in understanding this complicated membrane enzyme. We discuss mainly the mechanism of catalysis in the beta subunit of F(1) sector and roles of the gamma subunit in energy coupling. The gamma-subunit rotation during catalysis is also discussed. PMID- 10838045 TI - Molecular mechanisms of rotational catalysis in the F(0)F(1) ATP synthase. AB - Rotation of the F(0)F(1) ATP synthase gamma subunit drives each of the three catalytic sites through their reaction pathways. The enzyme completes three cycles and synthesizes or hydrolyzes three ATP for each 360 degrees rotation of the gamma subunit. Mutagenesis studies have yielded considerable information on the roles of interactions between the rotor gamma subunit and the catalytic beta subunits. Amino acid substitutions, such as replacement of the conserved gammaMet 23 by Lys, cause altered interactions between gamma and beta subunits that have dramatic effects on the transition state of the steady state ATP synthesis and hydrolysis reactions. The mutations also perturb transmission of specific conformational information between subunits which is important for efficient conversion of energy between rotation and catalysis, and render the coupling between catalysis and transport inefficient. Amino acid replacements in the transport domain also affect the steady state catalytic transition state indicating that rotation is involved in coupling to transport. PMID- 10838046 TI - ATP synthase: what we know about ATP hydrolysis and what we do not know about ATP synthesis. AB - In ATP synthase, X-ray structures, demonstration of ATP-driven gamma-subunit rotation, and tryptophan fluorescence techniques to determine catalytic site occupancy and nucleotide binding affinities have resulted in pronounced progress in understanding ATP hydrolysis, for which a mechanism is presented here. In contrast, ATP synthesis remains enigmatic. The molecular mechanism by which ADP is bound in presence of a high ATP/ADP concentration ratio is a fundamental unknown; similarly P(i) binding is not understood. Techniques to measure catalytic site occupancy and ligand binding affinity changes during net ATP synthesis are much needed. Relation of these parameters to gamma-rotation is a further goal. A speculative model for ATP synthesis is offered. PMID- 10838047 TI - The participation of metals in the mechanism of the F(1)-ATPase. AB - The Mg(2+) cofactor of the F(1)F(0) ATP synthase is required for the asymmetry of the catalytic sites that leads to the differences in affinity for nucleotides. Vanadyl (V(IV)=O)(2+) is a functional surrogate for Mg(2+) in the F(1)-ATPase. The (51)V-hyperfine parameters derived from EPR spectra of VO(2+) bound to specific sites on the enzyme provide a direct probe of the metal ligands at each site. Site-directed mutations of residues that serve as metal ligands were found to cause measurable changes in the (51)V-hyperfine parameters of the bound VO(2+), thereby providing a means by which metal ligands were identified in the functional enzyme in several conformations. At the low-affinity catalytic site comparable to beta(E) in mitochondrial F(1), activation of the chloroplast F(1) ATPase activity induces a conformational change that inserts the P-loop threonine and catch-loop tyrosine hydroxyl groups into the metal coordination sphere thereby displacing an amino group and the Walker homology B aspartate. Kinetic evidence suggests that coordination of this tyrosine by the metal when the empty site binds substrate may provide an escapement mechanism that allows the gamma subunit to rotate and the conformation of the catalytic sites to change, thereby allowing rotation only when the catalytic sites are filled. In the high-affinity conformation analogous to the beta(DP) site of mitochondrial F(1), the catch-loop tyrosine has been displaced by carboxyl groups from the Walker homology B aspartate and from betaE197 in Chlamydomonas CF(1). Coordination of the metal by these carboxyl groups contributes significantly to the ability of the enzyme to bind the nucleotide with high affinity. PMID- 10838048 TI - Important subunit interactions in the chloroplast ATP synthase. AB - General structural features of the chloroplast ATP synthase are summarized highlighting differences between the chloroplast enzyme and other ATP synthases. Much of the review is focused on the important interactions between the epsilon and gamma subunits of the chloroplast coupling factor 1 (CF(1)) which are involved in regulating the ATP hydrolytic activity of the enzyme and also in transferring energy from the membrane segment, chloroplast coupling factor 0 (CF(0)), to the catalytic sites on CF(1). A simple model is presented which summarizes properties of three known states of activation of the membrane-bound form of CF(1). The three states can be explained in terms of three different bound conformational states of the epsilon subunit. One of the three states, the fully active state, is only found in the membrane-bound form of CF(1). The lack of this state in the isolated form of CF(1), together with the confirmed presence of permanent asymmetry among the alpha, beta and gamma subunits of isolated CF(1), indicate that ATP hydrolysis by isolated CF(1) may involve only two of the three potential catalytic sites on the enzyme. Thus isolated CF(1) may be different from other F(1) enzymes in that it only operates on 'two cylinders' whereby the gamma subunit does not rotate through a full 360 degrees during the catalytic cycle. On the membrane in the presence of a light-induced proton gradient the enzyme assumes a conformation which may involve all three catalytic sites and a full 360 degrees rotation of gamma during catalysis. PMID- 10838049 TI - The IF(1) inhibitor protein of the mitochondrial F(1)F(0)-ATPase. AB - Recent studies on the IF(1) inhibitor protein of the mitochondrial F(1)F(0) ATPase from molecular biochemistry to possible pathophysiological roles are reviewed. The apparent mechanism of IF(1) inhibition of F(1)F(0)-ATPase activity and the biophysical conditions that influence IF(1) activity are summarized. The amino acid sequences of human, bovine, rat and murine IF(1) are compared and domains and residues implicated in IF(1) function examined. Defining the minimal inhibitory sequence of IF(1) and the role of conserved histidines and conformational changes using peptides or recombinant IF(1) is reviewed. Luft's disease, a mitochondrial myopathy where IF(1) is absent, is described with respect to IF(1) relevance to mitochondrial bioenergetics and clinical observations. The possible pathophysiological role of IF(1) in conserving ATP under conditions where cells experience oxygen deprivation (tumor growth, myocardial ischemia) is evaluated. Finally, studies attempting to correlate IF(1) activity to ATP conservation in myocardial ischemic preconditioning are compared. PMID- 10838050 TI - The second stalk of Escherichia coli ATP synthase. AB - Two stalks link the F(1) and F(0) sectors of ATP synthase. The central stalk contains the gamma and epsilon subunits and is thought to function in rotational catalysis as a rotor driving conformational changes in the catalytic alpha(3)beta(3) complex. The two b subunits and the delta subunit associate to form b(2)delta, a second, peripheral stalk extending from the membrane up the side of alpha(3)beta(3) and binding to the N-terminal regions of the alpha subunits, which are approx. 125 A from the membrane. This second stalk is essential for binding F(1) to F(0) and is believed to function as a stator during rotational catalysis. In vitro, b(2)delta is a highly extended complex held together by weak interactions. Recent work has identified the domains of b which are essential for dimerization and for interaction with delta. Disulphide cross linking studies imply that the second stalk is a permanent structure which remains associated with one alpha subunit or alphabeta pair. However, the weak interactions between the polypeptides in b(2)delta pose a challenge for the proposed stator function. PMID- 10838051 TI - The ATP synthase of Escherichia coli: structure and function of F(0) subunits. AB - In this review we discuss recent work from our laboratory concerning the structure and/or function of the F(0) subunits of the proton-translocating ATP synthase of Escherichia coli. For the topology of subunit a a brief discussion gives (i) a detailed picture of the C-terminal two-thirds of the protein with four transmembrane helices and the C terminus exposed to the cytoplasm and (ii) an evaluation of the controversial results obtained for the localization of the N terminal region of subunit a including its consequences on the number of transmembrane helices. The structure of membrane-bound subunit b has been determined by circular dichroism spectroscopy to be at least 75% alpha-helical. For this purpose a method was developed, which allows the determination of the structure composition of membrane proteins in proteoliposomes. Subunit b was purified to homogeneity by preparative SDS gel electrophoresis, precipitated with acetone, and redissolved in cholate-containing buffer, thereby retaining its native conformation as shown by functional coreconstitution with an ac subcomplex. Monoclonal antibodies, which have their epitopes located within the hydrophilic loop region of subunit c, and the F(1) part are bound simultaneously to the F(0) complex without an effect on the function of F(0), indicating that not all c subunits are involved in F(1) interaction. Consequences on the coupling mechanism between ATP synthesis/hydrolysis and proton translocation are discussed. PMID- 10838052 TI - Operation of the F(0) motor of the ATP synthase. AB - ATP, the universal carrier of cell energy is manufactured from ADP and phosphate by the enzyme ATP synthase using the energy stored in a transmembrane ion gradient. The two components of the ion gradient (DeltapH or DeltapNa(+)) and the electrical potential difference Deltapsi are thermodynamically but not kinetically equivalent. In contrast to accepted wisdom, the electrical component is kinetically indispensable not only for bacterial ATP synthases but also for that from chloroplasts. Recent biochemical studies with the Na(+)-translocating ATP synthase of Propionigenium modestum have given a good idea of the ion translocation pathway in the F(0) motor. Taken together with biophysical data, the operating principles of the motor have been delineated. PMID- 10838053 TI - Structural interpretations of F(0) rotary function in the Escherichia coli F(1)F(0) ATP synthase. AB - F(1)F(0) ATP synthases are known to synthesize ATP by rotary catalysis in the F(1) sector of the enzyme. Proton translocation through the F(0) membrane sector is now proposed to drive rotation of an oligomer of c subunits, which in turn drives rotation of subunit gamma in F(1). The primary emphasis of this review will be on recent work from our laboratory on the structural organization of F(0), which proves to be consistent with the concept of a c(12) oligomeric rotor. From the NMR structure of subunit c and cross-linking studies, we can now suggest a detailed model for the organization of the c(12) oligomer in F(0) and some of the transmembrane interactions with subunits a and b. The structural model indicates that the H(+)-carrying carboxyl of subunit c is located between subunits of the c(12) oligomer and that two c subunits pack in a front-to-back manner to form the proton (cation) binding site. The proton carrying Asp61 side chain is occluded between subunits and access to it, for protonation and deprotonation via alternate entrance and exit half-channels, requires a swiveled opening of the packed c subunits and stepwise association with different transmembrane helices of subunit a. We suggest how some of the structural information can be incorporated into models of rotary movement of the c(12) oligomer during coupled synthesis of ATP in the F(1) portion of the molecule. PMID- 10838054 TI - The structure of the H(+)-ATP synthase from chloroplasts and its subcomplexes as revealed by electron microscopy. AB - The electron microscopic data available on CF(0)F(1) and its subcomplexes, CF(0), CF(1), subunit III complex are collected and the CF(1) data are compared with the high resolution structure of MF(1). The data are based on electron microscopic investigation of negatively stained isolated CF(1), CF(0)F(1) and subunit III complex. In addition, two-dimensional crystals of CF(0)F(1) and CF(0)F(1) reconstituted liposomes were investigated by cryo-electron microscopy. Progress in the interpretation of electron microscopic data from biological samples has been made with the introduction of image analysis. Multi-reference alignment and classification of images have led to the differentiation between different conformational states and to the detection of a second stalk. Recently, the calculation of three-dimensional maps from the class averages led to the understanding of the spatial organisation of the enzyme. Such three-dimensional maps give evidence of the existence of a third connection between the F(0) part and F(1) part. PMID- 10838055 TI - Molecular models of the structural arrangement of subunits and the mechanism of proton translocation in the membrane domain of F(1)F(0) ATP synthase. AB - Subunit c of the proton-transporting ATP synthase of Escherichia coli forms an oligomeric complex in the membrane domain that functions in transmembrane proton conduction. The arrangement of subunit c monomers in this oligomeric complex was studied by scanning mutagenesis. On the basis of these studies and structural information on subunit c, different molecular models for the potential arrangement of monomers in the c-oligomer are discussed. Intersubunit contacts in the F(0) domain that have been analysed in the past by chemical modification and mutagenesis studies are summarised. Transient contacts of the c-oligomer with subunit a might play a crucial role in the mechanism of proton translocation. Schematic models presented by several authors that interpret proton transport in the F(0) domain by a relative rotation of the c-subunit oligomer against subunit a are reviewed against the background of the molecular models of the oligomer. PMID- 10838056 TI - Insights into ATP synthase assembly and function through the molecular genetic manipulation of subunits of the yeast mitochondrial enzyme complex. AB - Development of an increasingly detailed understanding of the eucaryotic mitochondrial ATP synthase requires a detailed knowledge of the stoichiometry, structure and function of F(0) sector subunits in the contexts of the proton channel and the stator stalk. Still to be resolved are the precise locations and roles of other supernumerary subunits present in mitochondrial ATP synthase complexes, but not found in the bacterial or chloroplast enzymes. The highly developed system of molecular genetic manipulation available in the yeast Saccharomyces cerevisiae, a unicellular eucaryote, permits testing for gene function based on the effects of gene disruption or deletion. In addition, the genes encoding ATP synthase subunits can be manipulated to introduce specific amino acids at desired positions within a subunit, or to add epitope or affinity tags at the C-terminus, enabling questions of stoichiometry, structure and function to be addressed. Newly emerging technologies, such as fusions of subunits with GFP are being applied to probe the dynamic interactions within mitochondrial ATP synthase, between ATP synthase complexes, and between ATP synthase and other mitochondrial enzyme complexes. PMID- 10838057 TI - Organisation of the yeast ATP synthase F(0):a study based on cysteine mutants, thiol modification and cross-linking reagents. AB - A topological study of the yeast ATP synthase membranous domain was undertaken by means of chemical modifications and cross-linking experiments on the wild-type complex and on mutated enzymes obtained by site-directed mutagenesis of genes encoding ATP synthase subunits. The modification by non-permeant maleimide reagents of the Cys-54 of mutated subunit 4 (subunit b), of the Cys-23 in the N terminus of subunit 6 (subunit a) and of the Cys-91 in the C-terminus of mutated subunit f demonstrated their location in the mitochondrial intermembrane space. Near-neighbour relationships between subunits of the complex were demonstrated by means of homobifunctional and heterobifunctional reagents. Our data suggest interactions between the first transmembranous alpha-helix of subunit 6, the two hydrophobic segments of subunit 4 and the unique membrane-spanning segments of subunits i and f. The amino acid residue 174 of subunit 4 is close to both oscp and the beta-subunit, and the residue 209 is close to oscp. The dimerisation of subunit 4 in the membrane revealed that this component is located in the periphery of the enzyme and interacts with other ATP synthase complexes. PMID- 10838058 TI - A model for the structure of subunit a of the Escherichia coli ATP synthase and its role in proton translocation. AB - Most of what is known about the structure and function of subunit a, of the ATP synthase, has come from the construction and isolation of mutations, and their analysis in the context of the ATP synthase complex. Three classes of mutants will be considered in this review. (1) Cys substitutions have been used for structural analysis of subunit a, and its interactions with subunit c. (2) Functional residues have been identified by extensive mutagenesis. These studies have included the identification of second-site suppressors within subunit a. (3) Disruptive mutations include deletions at both termini, internal deletions, and single amino acid insertions. The results of these studies, in conjunction with information about subunits b and c, can be incorporated into a model for the mechanism of proton translocation in the Escherichia coli ATP synthase. PMID- 10838059 TI - F(0)F(1)-ATP synthase: general structural features of 'ATP-engine' and a problem on free energy transduction. PMID- 10838060 TI - Reverse engineering a protein: the mechanochemistry of ATP synthase. AB - ATP synthase comprises two rotary motors in one. The F(1) motor can generate a mechanical torque using the hydrolysis energy of ATP. The F(o) motor generates a rotary torque in the opposite direction, but it employs a transmembrane proton motive force. Each motor can be reversed: The F(o) motor can drive the F(1) motor in reverse to synthesize ATP, and the F(1) motor can drive the F(o) motor in reverse to pump protons. Thus ATP synthase exhibits two of the major energy transduction pathways employed by the cell to convert chemical energy into mechanical force. Here we show how a physical analysis of the F(1) and F(o) motors can provide a unified view of the mechanochemical principles underlying these energy transducers. PMID- 10838061 TI - Entorhinal cortex pre-alpha cell clusters in schizophrenia: quantitative evidence of a developmental abnormality. AB - BACKGROUND: Previous studies using semiquantitative or qualitative techniques demonstrated abnormalities of positioning of clusters of neurons (pre-alpha cells) in the entorhinal cortex in schizophrenia, suggesting a developmental mechanism could contribute to the illness. Recent quantitative studies of laminar thickness and laminar cell counts have been less consistent, and several failed to replicate the finding. However, none of the quantitative studies focused on the position of the pre-alpha cell clusters. METHODS: To study pre-alpha cell position in detail, we examined the entorhinal cortex in serial sections from 21 control and 19 schizophrenic brains. Cluster position relative to the gray-white matter junction and cluster size were measured. RESULTS: Quantitative assessment of 1991 clusters indicated clusters were positioned relatively closer to the gray white matter junction in the anterior half of schizophrenic entorhinal cortices. In addition, the size of clusters in males with schizophrenia was reduced. CONCLUSIONS: These results support the model of schizophrenia as an illness in which brain development is impaired. The findings in males with schizophrenia may indicate the presence of more severe pathology, or an additional pathogenic mechanism. PMID- 10838062 TI - Reduced number of mediodorsal and anterior thalamic neurons in schizophrenia. AB - BACKGROUND: The thalamus is a brain region of interest in the study of schizophrenia because it provides critical input to brain regions such as the prefrontal, cingulate, and temporal cortices, where abnormalities have been repeatedly observed in patients with schizophrenia. Postmortem anatomic studies have rarely investigated the thalamus in this population. METHODS: Postmortem tissue was obtained from the left hemisphere of eight male schizophrenic patients and eight male age-matched control subjects. The optical dissector stereologic procedure was used to count neurons in the mediodorsal (MD) and anteroventral/anteromedial (AV/AM) nuclei of the thalamus. RESULTS: The number of neurons and volume of the MD were significantly reduced by 35% and 24%, respectively. The MD cell number reduction was a consistent finding; every control subject had more and every schizophrenic subject had fewer than 3.5 million neurons. Neuron number was also significantly reduced (16%) in the AV/AM nuclei. CONCLUSIONS: The present data indicate that schizophrenia is associated with robust reductions in nerve cell numbers in nuclei that communicate with the prefrontal cortex and limbic system. These thalamic anatomic deficits may be responsible, in part, for previous reports of such prefrontal cortical abnormalities as reduced synaptic density, reduced volume, and metabolic hypofunction. PMID- 10838063 TI - Reduced phosphodiesters and high-energy phosphates in the frontal lobe of schizophrenic patients: a (31)P chemical shift spectroscopic-imaging study. AB - BACKGROUND: (31)Phosphorous magnetic resonance spectroscopy has been widely used to evaluate schizophrenic patients in comparison to control subjects, because it allows the investigation of both phospholipid and energy metabolism in vivo; however, the results achieved so far are inconsistent. Chemical shift imaging (CSI) has the advantage that instead of only one or a few preselected voxels the tissue of a whole brain slice can be examined. The aim of the present investigation was to determine whether the results of previous studies of our group, showing that phosphodiesters (PDE) are decreased in the frontal lobe of schizophrenic patients as compared to control subjects, might be confirmed in an independent unmedicated patient sample using the CSI technique. METHODS: A carefully selected new cohort including 11 neuroleptic-free schizophrenic patients and 11 age- and gender-matched healthy control subjects was recruited. CSI was applied and an innovative analysis method for CSI data based on a general linear model was used. RESULTS: PDE, phosphocreatine, and adenosine triphosphate (ATP) were found to be significantly decreased in the frontal lobe of patients with schizophrenia. CONCLUSIONS: Because PDE was decreased in schizophrenic patients, the membrane phospholipid hypothesis of schizophrenia could not be corroborated. Further results indicate decreased ATP production in the frontal lobe of patients with schizophrenia. PMID- 10838064 TI - Differential preservation of cognitive functions in geriatric patients with lifelong chronic schizophrenia: less impairment in reading compared with other skill areas. AB - BACKGROUND: Our study examined the differential performance of cognitive skills in geriatric, cognitively impaired schizophrenic patients (n = 165) with a lengthy course of institutional stay and a poor overall functional outcome. Their relative deficits were compared with a sample of healthy elderly individuals. METHODS: Schizophrenic patients were matched one-to-one with healthy individuals of the same age and education and compared on a number of measures of cognitive functioning. The schizophrenic patients' old-learning performance was also compared with their educational level only. RESULTS: Mini-Mental State Examination (Folstein et al 1975) scores of the patients were in the moderately demented range (M = 20.36), and these patients underperformed healthy control subjects by more than 1 to slightly less than 3 standard deviations on measures of memory, praxis, and verbal skills. Wide Range Achievement Test-Revised word recognition reading scores were found to be at the 10th-grade level, although the patients on average had completed 11 years of formal education. CONCLUSIONS: These results suggest that even in schizophrenic patients with significant cognitive impairment, reading scores are relatively consistent with educational attainment. These data indicate that poor performance on measures of cognitive functioning in this population does not necessarily occur on measures of old learning. PMID- 10838065 TI - The P50 auditory event-evoked potential in adult attention-deficit disorder: comparison with schizophrenia. AB - BACKGROUND: Attention-deficit/hyperactivity disorder (ADHD) and schizophrenia are both conceptualized as disorders of attention. Failure to inhibit the P50 auditory event-evoked response, extensively studied in schizophrenia, could also occur in ADHD patients, if these two illnesses have common underlying neurobiological substrates. METHODS: This study examined the inhibition of the P50 auditory event-evoked potential in 16 unmedicated adults with ADHD, 16 schizophrenic outpatients, and 16 normal control subjects. Auditory stimuli were presented in a paired stimulus, conditioning-testing paradigm. RESULTS: The amplitude of initial or conditioning P50 response did not differ between the three groups; however, significant effects of psychiatric diagnosis on the amplitude of the test response and the ratio of the test to the conditioning response amplitudes were observed. Schizophrenic patients' P50 ratios and test amplitudes were higher than both the ADHD and normal groups. CONCLUSIONS: Adults with ADHD do not have the inhibitory deficit seen in patients with schizophrenia, suggesting that the mechanism of attentional disturbance in the two illnesses may be fundamentally different. PMID- 10838066 TI - Impaired modulation of the saccadic contingent negative variation preceding antisaccades in schizophrenia. AB - BACKGROUND: The contingent negative variation (CNV) is considered to reflect prefrontal functioning and can be observed before manual and ocular motor responses. Schizophrenic patients exhibit reduced CNV amplitudes in tasks requiring manual motor responses. A number of studies has also found normal prosaccades, but delayed antisaccades and an augmented rate of erroneous prosaccades during the antisaccade task in schizophrenia. In this study we examined the CNV during pro- and antisaccade tasks in schizophrenic patients and healthy control subjects. METHODS: Data of 17 medicated schizophrenics (ICD-10, F20) and 18 control subjects, matched with patients for age, gender, and education were analyzed. Horizontal pro- and antisaccades were elicited in four blocks, each consisting of 80 trials. Electroencephalogram was recorded from 32 channels with a DC amplifier. RESULTS: Patients exhibited delayed correct responses and more erroneous prosaccades during the antisaccade task than control subjects, but normal prosaccadic reaction times. In control subjects, the vertex predominant saccadic CNV was generally larger than in patients, and larger during the anti- than during the prosaccade task. This task-related amplitude augmentation was absent in patients. Analyses of additional components suggested specificity of impaired event-related potential modulation to the saccadic CNV. CONCLUSIONS: In accordance with the presumed prefrontal dysfunction, our results suggest deficient preparation and execution of antisaccades in schizophrenia. PMID- 10838067 TI - Muscle biopsy, macro EMG, and clinical characteristics in patients with schizophrenia. AB - BACKGROUND: In a previous study of motor unit properties in patients with schizophrenia, muscle fiber histologic and electrophysiologic abnormalities were observed. The present study was designed to compare patients with schizophrenia with healthy control subjects with regard to muscle fiber histology and motor unit function. A second objective was to relate these variables to clinical characteristics. METHODS: Twelve patients with first-episode schizophrenia and fifteen patients with chronic schizophrenia (DSM-III-R) and 27 matched control subjects were included in the study. Muscle biopsies were performed either in m. tibialis anterior or m. vastus lateralis. Electromyographic recordings (macro EMG) were made from the m. tibialis anterior motor units. Psychiatric ratings included the PANSS and extrapyramidal side effects. RESULTS: Seven of the muscle biopsy specimens from the patients and one from the control subjects were classified as abnormal (p =.049). The most frequent abnormality was atrophic muscle fibers. Eight patients and no control subjects exhibited pathological macro EMG (p =.032). The findings were present in chronic as well as in first episode patients with schizophrenia. CONCLUSIONS: In approximately 50% of the patients, neuromuscular abnormalities were found either in the muscle biopsy or the macro EMG investigations. The results indicate that either a common pathologic process or different pathological processes are at hand in the neuromuscular system in patients with schizophrenia. The findings are compatible with a disturbed cell membrane function. PMID- 10838068 TI - Olanzapine increases allopregnanolone in the rat cerebral cortex. AB - BACKGROUND: The neurosteroid allopregnanolone (3alpha-hydroxy-5alpha-pregnan-20 one) has anxiolytic and anticonvulsant properties, potentiating GABA(A) receptor chloride channel function with 20-fold higher potency than benzodiazepines. Behavioral studies demonstrate that olanzapine has anxiolyticlike properties in animals, but the mechanism responsible for these effects is not clear. We examined the effect of acute olanzapine administration on cerebral cortical allopregnanolone and its relationship to serum progesterone and corticosterone levels in rats. METHODS: Male Sprague-Dawley rats were habituated to intraperitoneal (IP) saline injection for 5 days. On the day of the experiment, rats were injected with olanzapine (0, 2.5, 5.0, or 10.0 mg/kg IP, 10-11 rats per condition). Rats were sacrificed 1 hour later, and cerebral cortical allopregnanolone levels and serum progesterone and corticosterone levels were measured by radioimmunoassay. RESULTS: Olanzapine increases cerebral cortical allopregnanolone up to fourfold, depending on dose. Positive correlations were observed between cerebral cortical allopregnanolone and serum progesterone levels and between cerebral cortical allopregnanolone and serum corticosterone levels. CONCLUSIONS: Olanzapine-induced increases in the potent GABA(A) receptor modulator allopregnanolone may alter GABAergic neurotransmission, possibly contributing to antipsychotic efficacy. If allopregnanolone alterations are linked to psychotic symptom relief, neurosteroids may represent molecules for pharmacologic intervention. PMID- 10838069 TI - Psychiatric sequelae of low birth weight at 11 years of age. AB - BACKGROUND: We examined the relationship between low birth weight (LBW) and psychiatric problems at age 11 years. METHODS: Random samples of 6-year-old LBW and normal birth weight (NBW) children from two socioeconomically disparate communities were identified, traced, and assessed. We targeted the 1983-1985 cohort of newborns who reached age 6 in 1990-1992, the scheduled period of fieldwork. Of the 1,095 in the target sample, 823 (75%) were assessed. Five years later, the sample was reassessed. Behavior problems were evaluated by standardized behavior problems scales rated by mothers and teachers. A multiple regression application that combines data from multiple informants was used. Prospective data were used to estimate the incidence of severe attention problems during the follow-up period. RESULTS: Information from mothers and teachers revealed that LBW was associated with an excess of attention problems at age 11 in the urban but not in the suburban children. In the urban setting, LBW children had a higher incidence of clinically significant attention problems than NBW children. Although LBW children scored higher than NBW children on externalizing problems, the effect was accounted for in large part by maternal smoking in pregnancy. CONCLUSIONS: The LBW-attention problems association observed in the urban community suggests an interaction between biologic vulnerability associated with premature birth and environmental risk associated with social disadvantage. Further research and replication are called for. PMID- 10838070 TI - Modulation of EWS/WT1 activity by the v-Src protein tyrosine kinase. AB - Desmoplastic small round cell tumor (DSRCT) is a malignant human cancer that is associated with a specific t(11;22) chromosome translocation, where 265 amino acids from the EWS amino-terminus are fused to the DNA binding domain of the WT1 tumor suppressor gene. We have noticed the presence of several SH3 interacting domains within the amino-terminus of EWS and have assessed the potential of EWS/WT1 to interact with such motifs. We find that EWS/WT1 can associate with the SH3 domain of several proteins, including v-Src. Ectopic expression of v-Src phosphorylates EWS/WT1 in vivo, as well as enhances the transactivation ability of the EWS amino-terminal domain. Structural alteration of the v-Src SH2 or SH3 domains produced mutants that could not interact with EWS/WT1 nor augment the transcriptional properties of EWS. Taken together, our results suggest the possibility that some transcriptional properties of EWS/WT1 may be regulated by a cytoplasmic signaling pathway. PMID- 10838071 TI - Differential incorporation of 1-beta-D-arabinofuranosylcytosine and 9-beta-D arabinofuranosylguanine into nuclear and mitochondrial DNA. AB - The anti-leukemic nucleoside analogs 1-beta-D-arabinofuranosylcytosine (araC) and 9-beta-D-arabinofuranosylguanine (araG) are dependent on intracellular phosphorylation for pharmacological activity. AraC is efficiently phosphorylated by deoxycytidine kinase (dCK). Although araG is phosphorylated by dCK in vitro, it is a preferred substrate of mitochondrial deoxyguanosine kinase. We have used autoradiography to show that araC was incorporated into nuclear DNA in Molt-4 and CEM T-lymphoblastoid cells as well as in Chinese hamster ovary cells. In contrast, araG was predominantly incorporated into mitochondrial DNA in the investigated cell lines, without detectable incorporation into nuclear DNA. These data suggest that the molecular targets of araG and araC may differ. PMID- 10838072 TI - Identification and light-induced expression of a novel gene of NADPH protochlorophyllide oxidoreductase isoform in Arabidopsis thaliana. AB - In Arabidopsis thaliana, we identified a novel gene of a NADPH protochlorophyllide oxidoreductase (POR) isoform, which catalyzes the light dependent protochlorophyllide a reduction in the chlorophyll (Chl) biosynthetic pathway. The deduced amino acid sequence of the novel POR isoform (PORC) showed significant identities ( approximately 75%) with the previously isolated two POR isoforms of A. thaliana. Contrasting with these POR isoforms, the PORC transcript increased in etiolated seedlings by illumination, and was dominantly expressed in immature and mature tissues. The present results demonstrated that Chl biosynthesis and chloroplast biogenesis in A. thaliana are controlled by three POR isoforms, which are differentially controlled by light and development. PMID- 10838074 TI - Phosphorylation-dependent activation of TAK1 mitogen-activated protein kinase kinase kinase by TAB1. AB - TAK1 is a mitogen-activated protein kinase kinase kinase (MAP3K) that is involved in the c-Jun N-terminal kinase/p38 MAPKs and NF-kappaB signaling pathways. Here, we characterized the molecular mechanisms of TAK1 activation by its specific activator TAB1. Autophosphorylation of two threonine residues in the activation loop of TAK1 was necessary for TAK1 activation. Association with TAK1 and induction of TAK1 autophosphorylation required the C-terminal 24 amino acids of TAB1, but full TAK1 activation required additional C-terminal Ser/Thr rich sequences. These results demonstrated that the association between the kinase domain of TAK1 and the C-terminal TAB1 triggered the phosphorylation-dependent TAK1 activation mechanism. PMID- 10838073 TI - A novel fluorescent probe diphenyl-1-pyrenylphosphine to follow lipid peroxidation in cell membranes. AB - Diphenyl-1-pyrenylphosphine (DPPP) was tested whether it could be used as a fluorescent probe to monitor lipid peroxidation in cell membranes. DPPP reacted with organic hydroperoxides and hydrogen peroxide stoichiometrically to give DPPP oxide (DPPP = O). DPPP incorporated into phosphatidylcholine liposomal membranes and polymorphonuclear leukocytes (PMNs) reacted with methyl linoleate hydroperoxide rapidly but not with hydrogen peroxide nor with tert-butyl hydroperoxide. This novel method revealed that lipid peroxidation proceeded within membranes of PMNs stimulated with phorbol 12-myristate 13-acetate, which is known to produce several kinds of free radicals. It was found that DPPP is a suitable fluorescent probe to monitor lipid peroxidation within cell membranes specifically. PMID- 10838075 TI - Synergistic activation of the mkp-1 gene by protein kinase A signaling and USF, but not c-Myc. AB - Mitogen-activated protein kinase phosphatase 1 (MKP-1) is negatively regulating mitogen-activated protein kinases and is therefore involved in early signaling processes. The expression of the mkp-1 gene is induced by growth factors and stress. The promoters of the human and murine mkp-1 genes contain several conserved DNA binding elements, including two cAMP response elements and an E box. We observed that the upstream stimulatory factor (USF), but not c-Myc, activated mkp-1. USF synergized with protein kinase A, thus providing evidence for a role of the E box, during signal-regulated stimulation of mkp-1. PMID- 10838076 TI - Role of the N-terminal region of staphylokinase (SAK): evidence for the participation of the N-terminal region of SAK in the enzyme-substrate complex formation. AB - Staphylokinase (SAK) forms an inactive 1:1 complex with plasminogen (PG), which requires both the conversion of PG to plasmin (Pm) to expose an active site in PG SAK activator complex and the amino-terminal processing of SAK to expose the positively charged (Lys-11) amino-terminus after removal of the 10 N-terminal amino acid residues from the full length protein. The mechanism by which the N terminal segment of SAK affects its PG activation capability was investigated by generating SAK mutants, blocked in the native amino-terminal processing site of SAK, and carrying an alteration in the placement of the positively charged amino acid residue, Lys-11, and further studying their interaction with PG, Pm, miniplasmin and kringle structures. A ternary complex formation between PG-SAK PG was observed when an immobilized PG-SAK binary complex interacted with free radiolabelled PG in a sandwich binding experiment. Formation of this ternary complex was inhibited by a lysine analog, 6-aminocaproic acid (EACA), in a concentration dependent manner, suggesting the involvement of lysine binding site(s) in this process. In contrast, EACA did not significantly affect the formation of binary complex formed by native SAK or its mutant derivatives. Furthermore, the binary (activator) complex formed between PG and SAK mutant, PRM3, lacking the N-terminal lysine 11, exhibited 3-4-fold reduced binding with PG, Pm or miniplasmin substrate during ternary complex formation as compared to native SAK. Additionally, activator complex formed with PRM3 failed to activate miniplasminogen and exhibited highly diminished activation of substrate PG. Protein binding studies indicated that it has 3-5-fold reduction in ternary complex formation with miniplasmin but not with the kringle structure. In aggregate, these observations provide experimental evidence for the participation of the N-terminal region of SAK in accession and processing of substrate by the SAK-Pm activator complex to potentiate the PG activation by enhancing and/or stabilizing the interaction of free PG. PMID- 10838077 TI - Inactivation of p53 and life span extension of human diploid fibroblasts by mot 2. AB - Normal human lung fibroblasts were transfected with expression plasmids encoding mot-2, an hsp70 family member that is associated with the immortal phenotype. After the empty vector-transfected controls had become senescent and positive for senescence-associated beta-galactosidase (SA-beta-gal), the mot-2-expressing cells continued to proliferate for an additional 12-18 population doublings and showed a young cell morphology and much lower SA-beta-gal activity. The tumor suppressor p53 was found to be transcriptionally inactivated in life span extended cells. We have thus shown for the first time that overexpression of mot 2 in normal human cells is able to permit their temporary escape from senescence. PMID- 10838078 TI - Covalently bound flavin in the NqrB and NqrC subunits of Na(+)-translocating NADH quinone reductase from Vibrio alginolyticus. AB - Na(+)-translocating NADH-quinone reductase (NQR) from the marine bacterium Vibrio alginolyticus is composed of six subunits (NqrA to NqrF). On SDS-PAGE of the purified complex, NqrB and NqrC subunits were found to give yellow-green fluorescent bands under UV illumination. Both the NqrB and NqrC, electroeluted from the gel, had an absorption maximum at 448 nm, and the fluorescence excitation maxima at 365 and 448 nm and the emission maximum at 514 nm. The electroeluted NqrB and NqrC, respectively, were identified from their N-terminal amino acid sequences. These results clearly indicated that the NqrB and NqrC subunits have covalently bound flavins. The two subunits were digested by protease and then the fluorescent peptide fragments were separated by a reversed phase high performance liquid chromatography. N-Terminal amino acid sequence analyses of the fluorescent peptides revealed that the flavin is linked to Thr 235 in the NqrB and Thr-223 in the NqrC subunits. This is the first example that the flavin is linked to a threonine residue. The amino acid sequence around the flavin-linked threonine was well conserved between NqrB and NqrC. Identification of the flavin group is in progress. PMID- 10838079 TI - Distinct carboxy-termini confer divergent characteristics to the mitogen activated protein kinase p38alpha and its splice isoform Mxi2. AB - The p38 family of mitogen-activated protein kinases is composed of several isoforms. Mxi2 is a splicing variant of p38alpha that harbors a unique carboxy terminus. Here we show that this sole divergence results in remarkable differences between Mxi2 and p38alpha. Mxi2 is distinctively activated by stress stimuli and potently activated by mitogens. Mxi2 affinity for bona fide p38 substrates is remarkably diminished and Mxi2 activity is largely unaffected by the phosphatase CL100. Also, Mxi2 sensitivity to inhibition by SB203580 is greatly reduced. Interestingly, we show that the p38 C-terminus is involved in conferring sensitivity to this compound. Overall, our results point to the p38 carboxy-terminus as a key determinant of the biochemical properties of this family of kinases. PMID- 10838080 TI - Augmentation of urea-synthetic capacity by inhibition of nitric oxide synthesis in butyrate-induced differentiated human hepatocytes. AB - We have recently developed an in vitro differentiation model of immortalized non transformed human hepatocytes using butyrate, and observed the induction of inducible NO synthase (iNOS). In this study, we analyzed the effect of NO on the urea-synthetic capacity of these cells. The inhibition of iNOS during butyrate treatment significantly increased the urea-synthetic capacity as compared to that of butyrate treatment alone, possibly through the further induction of ornithine transcarbamylase expression. Therefore, the inhibition of NO production might be useful for obtaining more differentiated hepatocytes in the process of in vitro induction of hepatocyte-specific differentiation. PMID- 10838081 TI - Phosphorylation of Hic-5 at tyrosine 60 by CAKbeta and Fyn. AB - Hic-5 is a CAKbeta-binding protein localized at focal adhesions. Here we show that overexpression of CAKbeta or Fyn, but not FAK, enhanced the tyrosine phosphorylation of coexpressed Hic-5 in COS-7 cells. These phosphorylations were further augmented by stimulating cells with osmotic stress. The Y60F mutant of Hic-5 was not phosphorylated, and Hic-5 phosphorylated on tyrosine 60 was bound specifically to the SH2 domain of Csk. Coexpression experiments revealed that the phosphorylation of Hic-5 by CAKbeta required the kinase activation of CAKbeta and binding of Hic-5 by CAKbeta. Specific phosphorylation of Hic-5 by CAKbeta and Fyn may activate a signaling pathway mediated by Hic-5. PMID- 10838082 TI - Priming of PC12 cells for semiquantitative microinjection studies involving Ras. AB - Nerve growth factor and activated Ras can induce differentiation of rat pheochromocytoma cells (PC12 cells) [Greene and Tischler (1976) Proc. Natl. Acad. Sci. USA 73, 2424-2428] from a chromaffin cell-like morphology into one that resembles sympathetic neurones. We developed a special treatment of PC12 cells which apparently synchronizes these cells such that they are more useful for semi quantitative microinjection studies for signal transduction pathways. This treatment leads to a faster and more reproducible differentiation which faithfully reproduces the involvement of Ras in the process and allows a comparison of the biological activity of different Ras mutants. It shows that G12V and Q61L oncogenic mutants are not equally potent in inducing differentiation. Partial loss-of-function mutations T35S, E37G and Y40C are inactive and even a triple combination of these does not restore full biological activity. PMID- 10838083 TI - Biochemical characterization of the Drosophila axin protein. AB - In the Wnt/Wingless pathway, accumulation of beta-catenin/Armadillo protein is a key regulatory step. Vertebrate Axin is a negative regulator of Wnt signaling, promoting glycogen synthase kinase-3beta-mediated phosphorylation of beta-catenin and thereby destabilizing beta-catenin. Using Drosophila cell culture systems, we demonstrated that a Drosophila homolog of Axin (Daxin) inhibits Wingless-induced Armadillo accumulation and Drosophila T-cell factor-dependent transcription induced by Wingless, Dishevelled, and Armadillo. The carboxy-terminal portion of Daxin is not essential for the inhibitory activity, but a mutant only consisting of this portion behaves as a dominant-negative protein. Moreover, interactions between Daxin and Zeste-white 3, Armadillo, Dishevelled, protein phosphatase 2A and Daxin itself were shown, providing direct evidence that Daxin is a scaffold protein in the Wingless pathway. PMID- 10838084 TI - The first peak of the UVB irradiation-dependent biphasic induction of vascular endothelial growth factor (VEGF) is due to phosphorylation of the epidermal growth factor receptor and independent of autocrine transforming growth factor alpha. AB - Ultraviolet B (UVB) irradiation, the major damaging component of sunlight, has earlier been reported to enhance cutaneous angiogenesis in chronically sun exposed skin. We herein provide first evidence for a biphasic induction of the vascular endothelial growth factor (VEGF) following UVB irradiation of the human epidermal cell line HaCaT. The first VEGF peak occurred on mRNA level at 1 h and on protein level at 4 h postirradiation and is fully mediated by the UVB dependent phosphorylation of the epidermal growth factor receptor, which subsequent to its phosphorylation also initiates at least in part the synthesis of transforming growth factor alpha that confers as shown previously the second late VEGF peak at 8 h on mRNA and at 24 h on protein level. PMID- 10838085 TI - The alpha2 and alpha5 integrin genes: identification of transcription factors that regulate promoter activity in epidermal keratinocytes. AB - We analysed the activity of the proximal promoters of the alpha2 and alpha5 integrin genes in human keratinocytes. An AP-1 site, found in the alpha5 but not the alpha2 promoter, bound c-Jun/c-Fos dimers and contributed strongly to promoter activity. Both promoters had a CCAAT/enhancer binding protein (C/EBP) binding site: the alpha5 C/EBP element enhanced activity, while the alpha2 site was a negative regulatory element. C/EBP overexpression repressed the activity of both promoters, but the effect was independent of occupancy of the identified C/EBP binding sites, suggesting interactions with additional transcription factors. We propose that upregulation of C/EBPs contributes to the inhibition of integrin transcription during keratinocyte terminal differentiation, while AP-1 factors play a role in the selective induction of the alpha5 gene during wound healing. PMID- 10838086 TI - Purification, structure determination and synthesis of covalitoxin-II, a short insect-specific neurotoxic peptide from the venom of the Coremiocnemis validus (Singapore tarantula). AB - Spider venoms contain toxins that specifically immobilize and kill insects. We report the purification and characterization of a new insect-specific toxin named covalitoxin-II (Cvtx-II; mass, 3406. 24+/-0.64), from Coremiocnemis validus (Singapore tarantula) venom. The complete 31 amino acid sequence of Cvtx-II has been determined and it shows less than 40% identity with spider toxins. However, Cvtx-II has conserved cystine motif analogous to other spider and omega conotoxins. Cvtx-II was chemically synthesized and identified with the native Cvtx-II. Synthetic Cvtx-II induced insect-specific non-lethal excitatory activity when injected into crickets, but not in cockroaches and mice. PMID- 10838087 TI - Human cytomegalovirus immediate early proteins upregulate endothelial p53 function. AB - Infected endothelial cells are found to be resistant to apoptosis possibly mediated by p53 cytoplasmic sequestration. We explored whether the immediate early 84 kDa protein (IE84) of cytomegalovirus (CMV) is responsible for p53 cytoplasmic sequestration. The endothelial cells were transfected with plasmids containing IE1 and 2 coding regions which are known to synthesize IE84 and 72 proteins. Our study found that p53 expression was significantly elevated in endothelial cells transfected with IE1 and 2 plasmids. However, p53 was only found in the nucleus rather than sequestered in the cytoplasm. We have demonstrated that IE84 and 72 are not responsible for p53 dysfunction caused by CMV infection, rather they upregulate p53 function and promote endothelial apoptosis. PMID- 10838088 TI - Protein kinases induced by osmotic stresses and elicitor molecules in tobacco cell suspensions: two crossroad MAP kinases and one osmoregulation-specific protein kinase. AB - Two protein kinases displaying mitogen-activated protein kinase (MAPK) properties are activated both by an hypoosmotic stress and by oligogalacturonides in tobacco cell suspensions [Cazale et al. (1999) Plant J. 19, 297-307]. Using specific antibodies, they were identified as the salicylic acid-induced protein kinase (SIPK) and wound-induced protein kinase (WIPK). The SIPK was also activated by an hyperosmotic stress, indicating that the same kinase may play a role both in hypo and hyperosmotic signalling pathways, in addition to its involvement in the transduction of elicitor signals. Using immunoprecipitation followed by two dimensional in-gel kinase assay, three molecular forms of the SIPK were observed, suggesting that additional modifications of the activated kinase may occur. In contrast to WIPK and SIPK, which are located at the crossroad of several transduction pathways initiated by elicitor or osmotic stimuli, a 44 kDa kinase, that would not belong to the MAPK family, appeared more specific to osmotic stress. PMID- 10838089 TI - Impaired stimulation of intestinal glucose absorption by portal insulin via hepatoenteral nerves in chronically ethanol-intoxicated rats. AB - In the isolated, jointly perfused small intestine and liver of rats insulin, infused into the portal vein, induced an increase in intestinal glucose absorption via hepatoenteral cholinergic nerves. The possible loss of function of these nerves due to ethanol-induced neuropathy was investigated with 6 weeks ethanol-fed rats. Portal insulin or arterial carbachol failed to increase intestinal glucose absorption but cAMP still did so. The intact stimulatory effect of cAMP indicated an undisturbed capacity of the enterocytes. The loss of action of portal insulin and of arterial carbachol can be explained by the impairment of the hepatoenteral nerves in line with an ethanol-induced neuropathy. PMID- 10838090 TI - EPR study of the dinuclear active copper site of tyrosinase from Streptomyces antibioticus. AB - The [Cu(I)-Cu(II)] half-met form of the dinuclear copper site of tyrosinase has been probed by continuous wave electron paramagnetic resonance (EPR) and hyperfine sublevel correlation (HYSCORE) spectroscopy in the presence and absence of inhibitors. In all cases the EPR spectrum is indicative of a d(x(2)-y(2)) ground state for the unpaired electron. From the cross-peaks observed in the HYSCORE spectra, proton hyperfine coupling constants were obtained that are compatible with a hydroxide ion in an equatorial coordination position of the paramagnetic copper. After changing the water solvent to D(2)O or after addition of the inhibitors p-nitrophenol or L-mimosine, the proton signals disappear. The relevance of these findings for understanding the catalytic cycle is discussed. PMID- 10838091 TI - Mutation of MyoD-Ser237 abolishes its up-regulation by c-Mos. AB - Recently we have shown that Mos could activate myogenic differentiation by promoting heterodimerisation of MyoD and E12 proteins. Here, we demonstrate that MyoD can be efficiently phosphorylated by in vitro kinase assay with purified Mos immunoprecipitated from transfected cells. Comparative two-dimensional tryptic phosphopeptide mapping combined with site-directed mutagenesis revealed that Mos phosphorylates MyoD on serine 237. Mutation of serine 237 to a non-phosphorylable alanine (MyoD-Ala237) abolished the positive regulation of MyoD by Mos following overexpression in proliferating 10T1/2 cells. Taken together, our data show that direct phosphorylation of MyoD-Ser237 by Mos plays a positive role in increasing MyoD activity during myoblast proliferation. PMID- 10838092 TI - Ferryl-oxo heme intermediate in the antimalarial mode of action of artemisinin. AB - Fourier transform infrared (FTIR) and resonance Raman (RR) spectroscopies have been employed to investigate the reductive cleavage of the O-O bond of the endoperoxide moiety of the antimalarial drug artemisinin and its analog trioxane alcohol by hemin dimer. We have recorded FTIR spectra in the nu(O-O) and nu(as)(Fe-O-Fe) regions of artemisinin and of the hemin dimer that show the cleavage of the endoperoxide and that of the hemin dimer, respectively. We observed similar results in the trioxane alcohol/hemin dimer reaction. The RR spectrum of the artemisinin/hemin dimer reaction displays a vibrational mode at 850 cm(-1) that shifts to 818 cm(-1) when the experiment is repeated with (18)O O(18) endoperoxide enriched trioxane alcohol. The frequency of this vibration and the magnitude of the (18)O-O(18) isotopic shift led us to assign the 850 cm(-1) mode to the Fe(IV) = O stretching vibration of a ferryl-xoxo heme intermediate that occurs in the artemisinin/hemin dimer and trioxane alcohol/hemin reactions. These results provide the first direct characterization of the antimalarial mode of action of artemisinin and its trioxane analog, and suggest that artemisinin appears to react with heme molecules that have been incorporated into hemozoin and subsequently the heme performs cytochrome P450-type chemistry. PMID- 10838093 TI - Identification of organic anion transporting polypeptide 4 (Oatp4) as a major full-length isoform of the liver-specific transporter-1 (rlst-1) in rat liver. AB - A novel organic anion transporting polypeptide (Oatp)4(1) was isolated from rat liver that is 35 amino acids longer than the reported rat liver specific organic anion transporter (rlst)-1 and exhibits a 64% amino acid sequence identity with the human OATP-C (LST-1/OATP2; gene symbol SLC21A6). When expressed in Xenopus laevis oocytes, Oatp4 (Slc21a10) mediated polyspecific uptake of a variety of organic anions including taurocholate (K(m) approximately 27 microM), bromosulfophthalein (K(m) approximately 1.1 microM) and steroid conjugates. Based on nuclease protection analysis Oatp4 appears to be the predominant transcript in rat liver indicating that rlst-1 plays a minor role in overall hepatic organic anion uptake. PMID- 10838094 TI - Varying effects of temperature, Ca(2+) and cytochalasin on fusion activity mediated by human immunodeficiency virus type 1 and type 2 glycoproteins. AB - We examined fusion mediated by the human immunodeficiency virus type 1 (HIV-1) and type 2 (HIV-2) envelope glycoproteins under various experimental conditions. Incubation of HeLa cells expressing HIV-2(ROD) and HIV-2(SBL/ISY) envelope glycoproteins with HeLa-CD4 target cells resulted in fusion at temperatures >/=25 degrees C whereas fusion with cells expressing HIV-1(Lai) occurred only at >/=31 degrees C. HIV-2 envelope glycoprotein-mediated fusion proceeded in the absence of Ca(2+) in the culture medium, whereas HIV-1 fusion required Ca(2+) ions for fusion. In contrast to HIV-2 envelope glycoprotein fusion, incubations in the presence of the 0.5 microM cytochalasin B completely inhibited HIV-1 envelope glycoprotein-mediated fusion. Our results suggest that in contrast to HIV-2, HIV 1 fusion is dependent on dynamic processes in the target membrane. PMID- 10838096 TI - Corrigendum to: key amino acids of vasopressin V1a receptor responsible for the species difference in the affinity of OPC-21268 (FEBS 23227). PMID- 10838095 TI - Does nitric oxide synthase catalyze the synthesis of superoxide? PMID- 10838105 TI - The therapeutics of melatonin: a paediatric perspective. AB - The production of melatonin by the pineal gland and its functions are considered, and then its possible uses in the treatment of children. Institutionalized children, and those with severe learning disorders, often have irregular sleep wake patterns, and there is evidence that melatonin can result in improvement to the benefit of both the child and the carers. The affected children can become less irritable, calmer, happier, and content. Also they may socialize better and become more attentive, with an improvement in their cognitive abilities. Another group of children who are likely to suffer from disturbed sleep are those who are visually handicapped. Melatonin given in the evening can improve their sleep patterns, and often their performance. No important side-effects have been reported. It is generally accepted that if a child is liable to epileptic seizures sleep deprivation may well exacerbate them. There is some evidence from clinical trials that in that event melatonin can be helpful. There are many other problems in which it is claimed that treatment with melatonin is justifiable. These are mentioned, but further confirmatory studies are needed in most of them. There is no doubt that melatonin can effect the circadian system, and shift the sleep-wake cycle; and that there are situations in which this can be desirable. PMID- 10838106 TI - Otocerebral anomalies associated with topical tretinoin use. AB - Topical preparations of tretinoin are used for the treatment of various skin conditions and for rejuvenation of the skin. Published information on pregnancy outcome following maternal exposure to topical tretinoin is limited to three case reports. We report a case of a patient with anomalies involving the ear and central nervous system with exposure to topical tretinoin during the first trimester. Though the potential link between the use of topical tretinoin and the existence of fetal malformations remains to be further documented by animal as well as epidemiological studies, we strongly recommend that the use of topical tretinoin during pregnancy should be discouraged, and effective contraception should be used in patients of childbearing age. PMID- 10838107 TI - Infantile bilateral striatal necrosis following measles. AB - A previously healthy 4-year-old boy presented with typical measles and demonstrated lesions confined to basal ganglia. The clinical symptoms were an abrupt onset, impaired consciousness and mutism, extrapyramidal signs and severe neurovegetative dysfunction. No modification of the cerebrospinal fluid was observed; laboratory tests were all normal with the exception of a positive serologic test for measles. Cranial magnetic resonance imaging showed abnormal signals in the striatum, affecting the putamen and the caudate nuclei bilaterally. Neurologic improvement occurred within 2 months, with regression of lesions on cranial imaging, suggesting that edema played an important role in the initial stage of the disease. PMID- 10838108 TI - On chronological changes in the basic EEG rhythm in persons with Down syndrome - with special reference to slowing of alpha waves. AB - The authors tried to know specificity of aging in persons with Down syndrome (DS) from the aspect of electroencephalograph (EEG) frequency changes through the cross-sectional and longitudinal studies, in comparison with normal persons as well as those with mentally retardation except the Down syndrome (non-DS MR). Subjects for a cross-sectional study were 265 persons with DS, 242 with non-DS MR and 239 healthy persons, and subjects for a follow-up study were 28 persons with DS and 14 with non-DS MR, whose EEGs were recorded repeatedly once a year during 8 or 9 years. Resting EEGs from the frontal, central and occipital regions were examined through power spectrum. In the cross-sectional study, the number of subjects with DS who showed dominant component within 8 Hz band of the basic rhythm reached maximum in its appearance rate at 40-44 years of age in the occipital area, but this slowing progressed already at 30-34 years of age. While in non-DS MR, the number of subjects who showed dominant component at 8 Hz reached maximum at 45-49 years of age, and this slowing of the basic rhythm was not so clear as in DS. In the follow-up study for subjects with DS, although the lowering in EEG frequency to 8 Hz took place in various years of age individually, earlier distinct decrease of the frequency was commonly noticed. These earlier steep lowering of EEG frequency was discussed in relation to the senile signs and to the decline of brain function referring to Alzheimer disease. PMID- 10838109 TI - Zonisamide - induced urinary lithiasis in patients with intractable epilepsy. AB - We report here three patients with intractable epilepsy who developed urinary lithiasis during zonisamide (ZNS) treatment. Abdominal pain due to left-sided hydronephrosis was the initial symptom in the first patient, and it was resolved after the excretion of a stone. The second patient, who had no specific symptoms, was found to have a thick sludge of calcium phosphate in the bladder when he suffered from aspiration pneumonia and dehydration. The third patient, who had a history of recurrent urinary obstruction, was also found to have a thick sludge of calcium oxalate in the bladder. The urinalysis of the three patients revealed alkaline urine and hypercalciuria. Although their urinary lithiasis was resolved by discontinuation of ZNS and supportive therapy, routine examination of urine parameters such as pH and sediments, and daily urine-output checks are thought to be necessary during treatment with ZNS, especially for patients who are bedridden for a long time and receive multiple antiepileptic drugs. PMID- 10838110 TI - Unique astrocytic inclusion in a 2 month-old baby showing Leigh-like brain lesions with lactic acidosis. AB - An unique cytoplasmic inclusion was found in astrocytes of a 2-month-old female baby who showed Leigh-like brain lesions with lactic acidosis, hypoglycemia and hepatomegaly. Although a defective enzyme was not determined, a metabolic disorder was suggested from clinicopathological observations. Symmetrically distributed lesions consisting of marked gliosis and proliferation of capillaries were observed in the basal ganglia, thalami and tegmentum. The astrocytic cytoplasmic inclusion was exclusively found in the cerebral and cerebellar white matter, where myelination was immature. The inclusion was round and eosinophilic, and positive for glial fibrillary acidic protein, vimentin, alphaB-crystallin, S 100 protein and microtubule associated protein 1B, immunohistochemically. An electron microscopic examination revealed an accumulation of intermediate filaments, ribosome and rough endoplasmic reticulum in the inclusion. The characteristic of this inclusion is different from that of other reported inclusions. The inclusion showed positive immunoreaction against CuZn superoxide dismutase, catalase, advanced glycation end-product and 4-hydroxy-2-nonenal antibodies, which suggest that oxidative stress is involved in the genesis of the inclusion. PMID- 10838111 TI - Midazolam as a first-line agent for status epilepticus in children. AB - Midazolam is a water-soluble benzodiazepine, and has recently emerged as a safe and effective treatment option after ordinary antiepileptic therapy in the management of status epilepticus. However, midazolam as a first-line agent for status epilepticus in children has not been fully investigated. Intravenous midazolam was used for status epilepticus in 27 children (38 convulsive episodes) from January 1997 to December 1999 in our hospital. Among them, 10 patients (16 convulsive episodes) were treated with intravenous midazolam as a first-line agent. The causes of the seizures varied. Midazolam was administered as an intravenous bolus dose (0.1-0.3 mg/kg), followed by continuous intravenous infusion (1-8 microg/kg per min). In all epileptic episodes but one, the seizures stopped within 1 min without any adverse effects. These results were compatible with the previously reported ones. It is important to terminate status epilepticus which can cause brain damage. Midazolam seems to be effective and safe as a first-line therapy for status epilepticus in children. PMID- 10838112 TI - Seizure-inducing paradoxical reaction to antiepileptic drugs. AB - We report on an 18-month-old girl with a seizure frequency of five/day, receiving an antiepileptic polytherapy consisting of primidone, clonazepam and phenytoin. Following discontinuation of clonazepam and primidone, the patient has been seizure-free under monotherapy for 2 years and shows marked developmental progress. Possible mechanisms of this paradoxical effect of antiepileptic drugs and the implications for antiepileptic therapy are discussed. PMID- 10838113 TI - Long-term medical, educational, and social prognoses of childhood-onset epilepsy: a population-based study in a rural district of Japan. AB - The long-term prognosis of childhood-onset epilepsy has rarely been studied in a general population. We examined the long-term medical, educational, and social outcomes in individuals with a history of childhood-onset epilepsy aged 20 years or older in a defined area of Japan. Furthermore, the patients' recognition of epilepsy as well as the parents' concerns about the prognosis of epilepsy were also surveyed. After a mean follow-up period of 18.9 years, we had sufficient data on 148 surviving patients aged 20-38 years (mean, 26. 0 years) and seven dead patients. The follow-up rate was 92.8%. In the overall group, the percentages of those who had had each of the following were as follows: (a) 5 year remission, 62.8%; (b) a relapse of seizures, 17.4%; (c) psychiatric complications, 2.7%; (d) mortality, 4.5%; (e) attendance at regular classes of an ordinary school during compulsory education, 71.6%; (f) entrance to high school, 65.5%; (g) employment, 67.4%; (h) marriage, 23.0%; and (i) acquisition of a driver's license, 54.7%. The educational and social variables of the control population were as follows: (e) 99.1, (f) 97.0, (g) 96.6, (h) 51.9, and (i) 94.8%. In the 99 patients of normal intelligence, the results of the same analysis were as follows: (a) 75.8, (b) 10.7, (c) 0, (d) 0.6, (e) 100, (f) 96.0, (g) 95.2, (h) 33.3, and (i) 77.8%, in contrast to the corresponding variables of the 49 patients with mental retardation, that is, (a) 36.7, (b) 44.4, (c) 8.2, (d) 12.2, (e) 14.3, (f) 6.1, (g) 20.4, (h) 2.0, and (i) 4.1%, respectively. The best predictors of seizure remission included an early response to therapy, a low frequency of seizures or an absence of status epilepticus prior to therapy, and normal mental development. As for the current awareness of epilepsy and its prognosis, nearly 40% of the patients did not know the true name of their illness, and the same proportion of parents were still anxious about the prognosis even if their children had been taken off medication. Our data show that the majority of patients with childhood-onset epilepsy have a favorable long term medical prognosis in terms of seizure remission and psychiatric complications, whereas they have a higher mortality rate and lower levels of educational background as well as employment and marital status compared with the general population. However, except for the low marriage rate for the younger age group and the low rate of holding a driver's license, patients of normal intelligence appear to have more favorable long-term prognoses in terms of educational and social aspects as well. In Japan, individuals with epilepsy may not seek a driver's license because of the law. PMID- 10838114 TI - Haloperidol-induced rhabdomyolysis without neuroleptic malignant syndrome in a handicapped child. AB - A 6-year-old severely handicapped boy with haloperidol-induced mild rhabdomyolysis without neuroleptic malignant syndrome is reported. He had suffered from frontal lobe epilepsy, severe mental retardation and involuntary movements. To reduce the involuntary movements, oral administration of haloperidol was started. Haloperidol was effective for the involuntary movements, and he became well. However, his urine was sometimes dark brown, and the myoglobin level in his urine was increased. After discontinuation of haloperidol, his urine never became dark brown. It was suspected that haloperidol might cause mild rhabdomyolysis without neuroleptic malignant syndrome. Neuroleptic malignant syndrome is a rare but severe adverse effect of neuroleptics, and is usually accompanied by rhabdomyolysis. Although there have been a few adult cases presenting rhabdomyolysis without neuroleptic malignant syndrome, this patient might be the first child case with such symptoms. We have to pay attention to this syndrome when using haloperidol even in children. PMID- 10838115 TI - Acute disseminated encephalomyelitis after live rubella vaccination. AB - We report here a case involving a 14-year-old boy who developed acute disseminated encephalomyelitis following live rubella vaccination. The patient became febrile and began to experience nuchal pain 16 days after the immunization. By 22 days after immunization, he experienced difficulty in walking. By 24 days, he had developed tetraparesis with retention of urine, and total sensory loss below the Th1 dermatomal level. He was febrile at this point and showed nuchal rigidity and Lhermitte's sign. Cerebrospinal fluid examination revealed elevated cell counts, protein level, and myelin basic protein. T2 weighted magnetic resonance imaging detected high intensity lesions in the bilateral cerebral white matter and cervical spinal cord. Following the administration of intravenous corticosteroids, the patient's clinical symptoms improved rapidly. PMID- 10838116 TI - Central core disease and congenital neuromuscular disease with uniform type 1 fibers in one family. AB - We report a family in which the father had central core disease and his son had congenital neuromuscular disease with uniform type 1 fibers. This is the first report of such a combination. Although they had no recognized mutation in the ryanodine receptor gene, it is highly likely that the son also had central core disease but without core structures. The absence of cores may be due to the muscle sample or the young age of the patient since core structures have been reported to increase with age. Although the prevalence of core structures in individual muscles is unknown, there is a possibility of sampling error. In some patients, congenital neuromuscular disease with uniform type 1 fibers is closely related to or identical with central core disease. PMID- 10838117 TI - Hypoxic-ischemic encephalopathy with cystic brain stem necroses and thalamic calcifications in a preterm twin. AB - A severe and rare ischemic brain lesion in a preterm twin boy is reported. The boy was born after two weeks of anhydramnios and amnionic infection at 24 weeks of gestation. Following a difficult Caesarean section and prolonged umbilical cord compression he developed prenatal acidosis with an umbilical cord pH of 6.96. At the age of 7 h, heart rate variability narrowed due to severely disturbed brain stem function and the patient developed clinical signs of hypoxic ischemic encephalopathy. Sonography demonstrated extensive symmetrical brain stem and basal ganglia lesions. After a prolonged comatose and apneic state, death occurred at the age of 25 days. Autopsy confirmed columnar bilateral cavitation of basal ganglia, diencephalon, brain stem and spinal gray matter, as well as focal calcifications in the palladium, thalamus, and brain stem. The findings highly resemble those observed after experimental or clinical cardiac arrest. PMID- 10838118 TI - Empty sella syndrome in nevoid basal cell carcinoma syndrome. AB - We reported the magnetic resonance imaging of four young patients (13 to 19 years) with nevoid basal cell carcinoma syndrome (NBCCS), which showed empty sella, agenesis of the corpus callosum and empty sella, an interhemispheric lipoma with callosal dysgenesis, and an arachnoid cyst in the posterior fossa, respectively. Calcification of the diaphragma sellae, which is a protective barrier against the pulsating action of the cerebrospinal fluid, may cause the empty sella in NBCCS. PMID- 10838119 TI - The oestrogenic potential of the phthalate esters. AB - The phthalate esters represent a class of chemicals used widely and diversely in industry. Concern that phthalates might be oestrogenic arose from observations that the diesters inhibited the binding of 17beta-estradiol to isolated oestrogen receptors and stimulated the expression of cellular oestrogen-sensitive endpoints (gene expression, mitosis) in vitro. However, conflicting results have been found in comparable studies, and those studies that have demonstrated oestrogen mimicry have generally done so at concentrations approaching, or above, the limit of water solubility for the phthalates. The monoesters (the primary metabolites of the diesters in vivo) are inactive in similar in vitro tests. Furthermore, the diesters have not shown any oestrogenic activity in numerous and diverse studies in vivo at doses eliciting systemic toxicity. Consequently, the oestrogenic activity of phthalates identified in in vitro studies is not relevant to humans or the environment. PMID- 10838120 TI - Detection of dopaminergic modulators in a tier I screening battery for identifying endocrine-active compounds (EACs). AB - Apomorphine (APO; D(2) receptor agonist), haloperidol (HAL; D(2) receptor antagonist), and reserpine (RES; a dopamine depletor that acts to lower brain dopamine levels by depleting central nervous system monoamines via disrupting storage vesicle function) have been examined in a Tier I screening battery, which has been designed to detect endocrine-active compounds (EACs). The Tier I battery incorporates two short-term in vivo tests (a 5-day ovariectomized female battery and a 15-day intact male battery using Sprague-Dawley rats) and an in vitro yeast transactivation system (YTS). In addition, two blood collection procedures were evaluated for their utility in detecting HAL-induced increases in serum prolactin (PRL) levels (i.e., the stress associated with each procedure). In the in vivo female battery, both HAL and RES increased serum PRL concentrations as expected, although the increase caused by RES was marginal. Increases in serum PRL levels are enhanced when daily dosages are administered via multiple-daily dosing of the test compound, which results in higher sustained blood levels of the test compounds. APO failed to decrease serum PRL concentrations in the female battery. In the in vivo male battery, HAL increased serum PRL concentrations as expected. However, APO and RES failed to affect serum PRL concentrations. The blood collection comparison experiment demonstrated that possible confounding of the data can occur with serum PRL concentrations when animals are exposed to stress. Basal levels of PRL were approximately fourfold higher in animals that were bled via the tail vein procedure when compared to PRL levels from animals that were bled under CO(2) anesthesia at euthanization. As a result of the higher basal PRL levels, the HAL-induced increase in serum PRL concentrations was completely attenuated in the tail-vein bled animals (1.3-fold). In contrast, HAL produced a fivefold increase in serum PRL in animals where blood was collected under CO(2) anesthesia at euthanization. Hence, collection of blood from animals under CO(2) anesthesia at euthanization is an acceptable approach for detection of compounds that increase PRL. In summary, HAL-like compounds would be identified in the Tier I male and female battery primarily via increased serum PRL concentrations. RES like compounds would be identified in the Tier I male battery via decreased gonadotropins and steroids and possibly in the Tier I female battery by a minimal increase in serum PRL concentrations. Compounds that produce a marginal increase in serum PRL when administered using single daily dosing can also be confirmed in an in vivo female battery with multiple dosing because this regimen increases the magnitude of the PRL increase. APO, a D(2) receptor agonist, was not detected in the in vivo male or female batteries, but in both instances the top dosage produced minimal decreases in body weight (99 to 96% of control). Hence, the proposed Tier I battery needs to be further evaluated with higher dosages of APO and other D(2) receptor agonists to determine whether it is capable of detecting such agents. PMID- 10838121 TI - Incorporation of S-9 activation into an ER-alpha transactivation assay. AB - We evaluated the feasibility of incorporating an exogenous metabolic activating system into an estrogen receptor-alpha transactivation assay. 17beta-estradiol (E2), and the proestrogenic pesticide methoxychlor (MXC) were evaluated for activity in the presence and absence of Aroclor-1254 induced rat liver S-9 fractions. Both E2 and MXC responded consistently in the assay with average EC(50) values of 9.6 x 10(-11) M and 1.2 x 10(-5) M, respectively. In the presence of a 0.1% S-9 fraction, the EC(50) for E2 was increased to 1.4 x 10(-9) M and that for MXC decreased to 4.9 x 10(-7) M, with both compounds demonstrating increased secondary metabolite formation as evidenced by HPLC analysis. Consistent with these data, metabolites of E2 and MXC exhibited decreased and increased potencies, respectively, in the assay system relative to the parent molecules. S-9 was compatible with the MCF-7 reporter assay and has the potential to enhance detection of proestrogenic materials. PMID- 10838122 TI - Functional expression of P-glycoprotein in primary cultures of human cytotrophoblasts and BeWo cells. AB - The objective of this study was to investigate the functional expression of the efflux transporter, P-glycoprotein (P-gp), in primary cultures of human cytotrophoblasts and BeWo cell monolayers. Uptake studies with primary cultures of human cytotrophoblasts or BeWo cells were conducted with calcein-AM and vinblastine (P-gp markers) or fluorescein (MRP marker) in the presence of specific P-gp or MRP inhibitors. Results showed that the accumulation of P-gp substrates calcein-AM and vinblastine by BeWo cells or primary cultures of human cytotrophoblasts was significantly enhanced in the presence of a typical P-gp inhibitor, cyclosporin-A, or other inhibitors such as quinidine, verapamil, and dipyridamole. MRP inhibitors had no effect on the accumulation of calcein or fluorescein by BeWo cells. Western blots confirmed the presence of multidrug resistant gene product 1 (MDR1) in both primary cultures of human cytotrophoblasts and BeWo cells. This study demonstrates functional P-gp in term human trophoblasts and further supports the use of primary cultures of human cytotrophoblasts and BeWo cells as in vitro models of the trophoblast to investigate mechanisms regulating drug distribution across the placenta. PMID- 10838123 TI - The antiestrogen ICI 182,780 abolishes developmental injury for murine embryos exposed in vitro to o,p'-DDT(1). AB - Previously, we reported that in vitro exposure of murine embryos to 0.1 microg/ml o,p'-DDT (an estrogenic pesticide) significantly reduced development to blastocyst and mean cell number per embryo, and increased percent cell death by 96 h of culture. The objective of the present study was to determine if developmental injury induced by o,p'-DDT resulted from estrogenic, antiestrogenic, or unrelated adverse biologic mechanisms. Toward this objective, pronuclear embryos from CD-1 mice were cultured 96 h in medium supplemented with 0.1% ethanol (control) or 0.1 microg/ml o,p'-DDT, 17beta-estradiol, or ICI 182,780 dissolved in ethanol as single agents or as paired mixtures. As single agents, development to blastocyst and mean cell numbers were significantly reduced and percent apoptosis was significantly increased for embryos cultured in the presence of o,p'-DDT or ICI 182,780. Development to blastocyst was significantly reduced for embryos cultured in the presence of 17beta-estradiol. Beneficial interaction occurred when the receptor antagonist ICI 182,780 was combined with either receptor agonist (o,p'-DDT or 17beta-estradiol). In contrast, interaction was not significant when the two agonists were combined. The results indicate that developmental injury due to the estrogenic pesticide o,p'-DDT was abolished by the addition of the receptor antagonist ICI 182,780 and not by the receptor agonist 17beta-estradiol. The findings underscore the utility of the model for uncovering mechanisms of developmental injury. PMID- 10838124 TI - Uteroplacental circulatory disturbance mediated by prostaglandin f2alpha in rats exposed to microwaves. hiro-n@po.incl.ne.jp. AB - To clarify the effects of microwaves on pregnancy, uterine or uteroplacental blood flow and endocrine and biochemical mediators, including corticosterone, estradiol, prostaglandin E(2) (PGE(2)), and prostaglandin F(2)alpha (PGF(2)alpha), were measured in rats exposed to continuous-wave (CW) microwave at 2 mW/cm(2) incident power density at 2450 MHz for 90 min. Colonic temperature in virgin and pregnant rats was not significantly altered by microwave treatment. Microwaves decreased uteroplacental blood flow and increased progesterone and PGF(2)alpha in pregnant, but not in virgin rats. Intraperitoneal (i.p.) administration of angiotensin II, a uteroplacental vasodilator, before microwave exposure prevented the reduction in uteroplacental blood flow and the increased progesterone and PGF(2)alpha in pregnant rats. Increased corticosterone and decreased estradiol during microwave exposure were observed independent of pregnancy and pretreatment with angiotensin II. These results suggest that microwaves (CW, 2 mW/cm(2), 2450 MHz) produce uteroplacental circulatory disturbances and ovarian and placental dysfunction during pregnancy, probably through nonthermal actions. The uteroplacental disturbances appear to be due to actions of PGF(2)alpha and may pose some risk for pregnancy. PMID- 10838125 TI - Developmental toxicity of acephate by gavage in mice. AB - Acephate (O,S - dimethyl acetyl phosphoramidothioate), an organophosphate insecticide, was evaluated for its potential to produce developmental toxicity in mice after oral administration. Pregnant ICR (CD-1) mice were given sublethal doses of 0 (distilled water), 7, 14, and 28 mg/kg/day acephate by gavage on Gestation Days 6 through 15. Maternal effects in the 28 mg/kg/day dose group included cholinergic signs, decreased body weight at 15 and 18 days of gestation, and decreased absolute and relative brain weight. Placental weight was also decreased and liver weight was increased in the high dose group. Absolute and relative brain weight was decreased in the 14 mg/kg/day group. No maternal effects were apparent in the 7 mg/kg/day dose group. Maternal exposure to acephate during organogenesis significantly affected the number of implantations, number of live fetuses, number of early resorptions, mean fetal weight, and the incidence of external and skeletal malformations in the 28 mg/kg/day dose group. No visceral malformations were observed. On the basis of the present results acephate showed maternal and developmental toxicity at 28 mg/kg/day. PMID- 10838126 TI - Interaction of estradiol and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in an ovulation model: evidence for systemic potentiation and local ovarian effects. AB - Immature rats were treated with estradiol cypionate, (ECP, 0, 0.1, 1, or 2 mg/kg s.c.) followed 24 h later by TCDD (0 or 10 microg/kg orally). Follicular development was induced with eCG [5 or 10 IU subcutaneously (s.c.)] followed by an ovulatory dose of hCG (10 IU s. c.). Inhibition of ovulation by TCDD was potentiated by ECP in hypophysectomized but not intact rats. Only hypophysectomized rats exposed systemically to TCDD and ECP exhibited weight loss. Pair feeding mimicked the combined effects of TCDD and ECP in hypophysectomized rats. In another experiment, intact rats received ECP s.c. (0 or 2 mg/kg) and TCDD into the ovarian bursa (0 or 250 ng). Another group of intact rats received TCDD orally (10 microg/kg) and ECP into the ovarian bursa (0 or 1.5 microg). Blockade of ovulation by systemic or local TCDD was alleviated by ECP pretreatment. Estrogen increased the systemic toxicity of TCDD in rats whereas antagonizing its direct ovarian effects. PMID- 10838127 TI - Effects of lonidamine on testicular and epididymal proteins in the rat. AB - The mechanism responsible for the antispermatogenic activity of lonidamine (LND) [1-(2,4-dichlorobenzyl)-1H-indazole-3-carboxylic acid], a drug with low systemic toxicity and lack of significant hormonal effects, is still unclear but may be related to alterations of Sertoli cell proteins. Here, we confirmed that a single oral dose of LND (100 mg/kg b.w.) to sexually mature Sprague-Dawley rats causes shrinkage and weight reduction of the testes after 48 h. These macroscopic changes correlated with histologic alterations revealed by light microscopy, consistent with partially reversible inhibition of spermatogenesis. When the testes and the epididymides of animals treated with or without LND were homogenized and analyzed by the Bradford assay, a significant increase of total protein content was observed after 24 and 48 h. When these homogenates were analyzed by concanavalin blotting, specific changes of the major rat macroglobulins, i.e. alpha(1)-inhibitor-3, alpha(2)-macroglobulin, and alpha(1) macroglobulin, were noted. In particular, LND caused a decrease of testicular alpha(1)-inhibitor-3, but not an increase of testicular alpha(2)-macroglobulin, indicating a mild local inflammatory response to the drug. PMID- 10838128 TI - 28-day toxicology test: indenopyridine RTI 4587-056 in male Sprague-Dawley rats. AB - The potential toxicity of RTI 4587-056, a hexahydroindenopyridine analog of SANDOZ 20-438, was examined in adult male Sprague-Dawley rats. Testicular, intestinal, and erythropoietic histology was assessed after 28 days of gavage treatment at 0, 10, and 100 mg/kg/day. During the first 10 days, dose-related clinical signs included mild to moderate lethargy shortly after dosing, lower consumption of feed and water, and body weight loss or decreased weight gain. Tolerance developed, such that lethargy disappeared and weight gains were equivalent to the control group during the second through fourth weeks. The compound did not affect intestinal epithelium or bone marrow. RTI 4587-056 was a highly effective antispermatogenic agent at both doses causing epididymal hypospermia and testicular atrophy. Based upon the Spermatogenic Index ratings, still lower doses would be effective male contraceptive agents. RTI 4587-056 has potential as a male contraceptive without overt side effects. Further testing is required. PMID- 10838129 TI - Octylphenol inhibits testosterone biosynthesis by cultured precursor and immature Leydig cells from rat testes. AB - 4-tert-octyphenol (OP) is a surfactant additive widely used in the manufacture of a variety of detergents and plastic products. OP has been reported to mimic the actions of estrogen in many cellular systems. The present studies evaluated the direct effects of OP on human chorionic gonadotropin (hCG)-stimulated testosterone biosynthesis by cultured precursor and immature Leydig cells from 23 day old (prepubertal) rats. Exposure to increasing OP concentrations (1 to 2000 nM) progressively decreased hCG-stimulated testosterone formation in both precursor and immature Leydig cells at higher OP concentrations (100 or 500 to 2000 nM). Testosterone levels were reduced approximately 30 to 70% below control at the highest concentration in both cell types. Similar reductions in testosterone associated with OP exposure were observed in cells stimulated with 1 mM 8-Br-cAMP, suggesting that the main actions of OP occur after the generation of cAMP. Increasing concentrations of 17beta-estradiol (1 to 1000 nM) had no effect on hCG-stimulated testosterone formation in both precursor and immature Leydig cells and the inclusion of 100 nM ICI 182,780, a pure estrogen antagonist, in precursor and immature Leydig cells exposed to OP and hCG, did not alter the inhibition by higher OP concentrations of testosterone formation in both cell types. These results suggest that OP is a hormonally active agent, but that some of its actions are distinct from those of 17beta-estradiol and are not mediated through the estrogen receptor alpha or beta pathway. To further localize the potential site(s) of action of OP, cultured precursor and immature Leydig cells were exposed to increasing concentrations of OP and hCG for 24 h. Next, fresh media containing 1 microM 22(R)-hydroxycholesterol, 1 microM pregnenolone, 1 microM progesterone, or 1 microM androstenedione was added, and the conversion of each substrate to testosterone was determined after incubation for 4 h. The conversion of androstenedione to testosterone was unaffected by exposure to OP, suggesting that the 17beta-hydroxysteroid dehydrogenase step is not inhibited. However, the conversion of 22(R)-hydroxycholesterol, pregnenolone and progesterone all were inhibited by prior exposure to OP and hCG. This finding suggests that the 17alpha-hydroxylase/c17-20-lyase step, which converts progesterone to androstenedione, is inhibited by OP, and that the cholesterol side-chain cleavage and 3beta-hydroxysteroid dehydrogenase -isomerase steps, which convert cholesterol to pregnenolone and pregnenolone to progesterone, respectively, are other potential sites of OP action. Because concomitant exposure to the antioxidants alpha-tocopherol or ascorbate did not alter the inhibition of testosterone formation by higher OP concentrations, it does not appear that OP is acting as a pseudosubstrate for the generation of free radicals, which can damage P450 enzymes. PMID- 10838130 TI - Involvement of DNA damage and repair in mutational spectra. PMID- 10838131 TI - Influence of DNA repair on mutation spectra in Salmonella. AB - This paper reviews the influence of DNA repair on spontaneous and mutagen-induced mutation spectra at the base-substitution (hisG46) and -1 frameshift (hisD3052) alleles present in strains of the Salmonella (Ames) mutagenicity assay. At the frameshift allele (mostly a CGCGCGCG target), DeltauvrB influences the frequency of spontaneous hotspot mutations (-CG), duplications, and deletions, and it also shifts the sites of deletions and duplications. Cells with pKM101+DeltauvrB spontaneously produce complex frameshifts (frameshifts with an adjacent base substitution). The spontaneous frequency of 1-base insertions or concerted (templated) mutations is unaffected by DNA repair, and neither mutation is inducible by mutagens. Glu-P-1, 1-nitropyrene (1NP), and 2-acetylaminofluorene (2AAF) induce only hotspot mutations and are unaffected by pKM101, whereas benzo(a)pyrene and 4-aminobiphenyl induce only hotspot in pKM101(-), and hotspot plus complex in pKM101(+). At the base-substitution allele (mostly a CC/GG target), the DeltauvrB allele increases spontaneous transitions in the absence of pKM101 and increases transversions in its presence. The frequency of suppressor mutations is decreased 4x by DeltauvrB, but increased 7. 5x by pKM101. Both repair factors cause a shift in the proportion of mutations to the second position of the CC/GG target. With UV light and gamma-rays, the DeltauvrB allele increases the proportion of transitions relative to transversions. pKM101 is required for mutagenesis by Glu-P-1 and 4-AB, and the types and positions of the substitutions are not altered by the addition of the DeltauvrB allele. Changes in DNA repair appear to cause more changes in spontaneous than in mutagen-induced mutation spectra at both alleles. There is a high correlation (r(2)=0.8) between a mutagen's ability to induce complex frameshifts and its relative base substitution/frameshift mutagenic potency. A mutagen induces the same primary class of base substitution in TA100 (DeltauvrB, pKM101) as it does in Escherichia coli, mammalian cells, or rodents as well as in the p53 gene of human tumors associated with exposure to that mutagen. Thus, a mutagen induces the same primary class of base substitution in most organisms, reflecting the conserved nature of DNA replication and repair processes. PMID- 10838132 TI - In vivo formation and repair of cyclobutane pyrimidine dimers and 6-4 photoproducts measured at the gene and nucleotide level in Escherichia coli. AB - In vivo formation and repair of the major UV-induced DNA photoproducts, cyclobutane pyrimidine dimers (CPDs) and 6-4 pyrimidine-pyrimidone photoproducts (6-4 PPs), have been examined at the gene and nucleotide level in Escherichia coli. Each type of DNA photoproduct has individually been studied using photoreactivation and two newly developed assays; the multiplex QPCR assay for damage detection at the gene level and the reiterative primer extension (PE) assay for damage detection at the nucleotide level. In the E. coli lacI and lacZ genes, CPDs and 6-4 PPs form in a 2:1 ratio, respectively, during UV irradiation. Repair of 6-4 PPs is more efficient than repair of CPDs since, on the average, 42% of 6-4 PPs are repaired in both genes in the first 40 min following 200 J/m(2) UV irradiation, while 1% of CPDs are repaired. The location, relative frequency of formation, and efficiency of repair of each type of photoproduct was examined in the first 52 codons of the E. coli lacI gene at the nucleotide level. Hotspots of formation were found for each type of lesion. Most photoproducts are at sites where both CPDs and 6-4 PPs are formed. Allowing 40 min of recovery following 200 J/m(2) shows that in vivo repair of 6-4 PPs is about fourfold more efficient than the repair of CPDs. Comparison of the lesion-specific photoproduct distribution of the lacI gene with a UV-induced mutation spectrum from wild-type cells shows that most mutational hotspots are correlated with sites of a majority of CPD formation. However, 6-4 PPs are also formed at some of these sites with relatively high frequency. This information, taken together with the observation that 6-4 PPs are repaired faster than CPDs, suggest that the cause of mutagenic hotspots in wild-type E. coli is inefficient repair of CPDs. PMID- 10838133 TI - Toward an understanding of the role of DNA adduct conformation in defining mutagenic mechanism based on studies of the major adduct (formed at N(2)-dG) of the potent environmental carcinogen, benzo[a]pyrene. AB - The process of carcinogenesis is initiated by mutagenesis, which often involves replication past damaged DNA. One question - what exactly is a DNA polymerase seeing when it incorrectly copies a damaged DNA base (e.g., inserting dATP opposite a dG adduct)? - has not been answered in any case. Herein, we reflect on this question, principally by considering the mutagenicity of one activated form of benzo[a]pyrene, (+)-anti-B[a]PDE, and its major adduct [+ta]-B[a]P-N(2)-dG. In previous work, [+ta]-B[a]P-N(2)-dG was shown to be capable of inducing>95% G-->T mutations in one sequence context (5'-TGC), and approximately 95% G-->A mutations in another (5'-AGA). This raises the question - how can a single chemical entity induce different mutations depending upon DNA sequence context? Our current working hypothesis is that adduct conformational complexity causes adduct mutational complexity, where DNA sequence context can affect the former, thereby influencing the latter. Evidence supporting this hypothesis was discussed recently (Seo et al., Mutation Res. [in press]). Assuming this hypothesis is correct (at least in some cases), one goal is to consider what these mutagenic conformations might be. Based on molecular modeling studies, 16 possible conformations for [+ta]-B[a]P-N(2)-dG are proposed. A correlation between molecular modeling and mutagenesis work suggests a hypothesis (Hypothesis 3): a base displaced conformation with the dG moiety of the adduct in the major vs. minor groove gives G-->T vs. G-->A mutations, respectively. (Hypothesis 4, which is a generalized version of Hypothesis 3, is also proposed, and can potentially rationalize aspects of both [+ta]-B[a]P-N(2)-dG and AP-site mutagenesis, as well as the so-called "A-rule".) Finally, there is a discussion of how conformational complexity might explain some unusual mutagenesis results that suggest [+ta] B[a]P-N(2)-dG can become trapped in different conformations, and why we think it makes sense to interpret adduct mutagenesis results by modeling ds-DNA (at least in some cases), even though the mutagenic event must occur at a ss/ds-DNA junction in the presence of a DNA polymerase. PMID- 10838134 TI - Mutation spectra in supF: approaches to elucidating sequence context effects. AB - Shuttle vectors carrying the supF suppressor tRNA gene were originally developed for mutagenesis experiments in primate and human cells. Since then, the supF gene has been used as a mutation reporter in other mammalian cells, yeast, Escherichia coli, and transgenic mice. The widespread use of the vector for studies of many DNA reactive agents has produced a large database of mutation spectra. These provide primary information on the kinds and distribution of mutations provoked by many agents and, in many instances, allow comparisons between related agents or the same agent in different cell backgrounds. In this review we will discuss some of these data with a primary focus on the interpretation of UV mutation spectra. We will also describe our development and application of custom supF marker genes as an approach to studying the effect of sequence context on mutation hotspots and cold spots. Our studies suggest that C-C photoproducts are not mutagenic in certain sequence contexts in which T-C photoproducts are mutation hotspots. In addition, we have found several examples of sequence context effects acting as much as 80 bases away from the site of mutation. We will consider some of the problems raised by these studies and the possible resolution of some of them offered by the newly discovered family of damage bypass DNA polymerases. PMID- 10838135 TI - Mutational spectra for polycyclic aromatic hydrocarbons in the supF target gene. AB - An SV40-based shuttle vector system was used to identify the types of mutational changes and the sites of mutation within the supF DNA sequence generated by the four stereoisomers of benzo[c]phenanthrene 3,4-dihydrodiol 1,2-epoxide (B[c]PhDE), by racemic mixtures of bay or fjord region dihydrodiol epoxides (DE) of 5-methylchrysene, of 5, 6-dimethylchrysene, of benzo[g]chrysene and of 7 methylbenz[a]anthracene and by two direct acting polycyclic aromatic hydrocarbon carcinogens, 7-bromomethylbenz[a]anthracene (7-BrMeBA) and 7-bromomethyl-12 methylbenz[a]anthracene (7-BrMe-12-MeBA). The results of these studies demonstrated that the predominant type of mutation induced by these compounds is the base substitution. The chemical preference for reaction at deoxyadenosine (dAdo) or deoxyguanosine (dGuo) residues in DNA, which is in general correlated with the spatial structure (planar or non-planar) of the reactive polycyclic aromatic hydrocarbon, is reflected in the preference for mutation at A&z.ccirf;T or G&z.ccirf;C pairs. In addition, if the ability to react with DNA in vivo is taken into account, the relative mutagenic potencies of the B[c]PhDE stereoisomers are consistent with the higher tumorigenic activity associated with non-planar polycyclic aromatic hydrocarbons and their extensive reaction with dAdo residues in DNA. Comparison of the types of mutations generated by polycyclic aromatic hydrocarbons and other bulky carcinogens in this shuttle vector system suggests that all bulky lesions may be processed by a similar mechanism related to that involved in replication past apurinic sites. However, inspection of the distribution of mutations over the target gene induced by the different compounds demonstrated that individual polycyclic aromatic hydrocarbons induce unique patterns of mutational hotspots within the target gene. A polymerase arrest assay was used to determine the sequence specificity of the interaction of reactive polycyclic aromatic hydrocarbons with the shuttle vector DNA. The results of these assays revealed a divergence between mutational hotspots and polymerase arrest sites for all compounds investigated, i.e., sites of mutational hotspots do not correspond to sites where high levels of adduct formation occur, and suggested that some association between specific adducts and sequence context may be required to constitute a premutagenic lesion. A site specific mutagenesis system employing a single-stranded vector (M13mp7L2) was used to investigate the mutational events a single benzo[a]pyrene or benzo[c]phenanthrene dihydrodiol epoxide-DNA adduct elicits within specific sequence contexts. These studies showed that sequence context can cause striking differences in mutagenic frequencies for given adducts. In addition, these sequence context effects do not originate only from nucleotides immediately adjacent to the adduct, but are also modulated by more distal nucleotides. The implications of these results for mechanisms of polycyclic aromatic hydrocarbon induced mutagenesis and carcinogenesis are discussed. PMID- 10838136 TI - Mutations induced by reactive nitrogen oxide species in the supF forward mutation assay. AB - Nitric oxide is an important bioregulatory molecule with a range of physiological functions. Nitric oxide can also react with oxygen species to produce a range of reactive nitrogen oxides that can damage DNA and lead to mutations of the DNA base sequence. The mutagenicity of a variety of reactive nitrogen oxide species and related DNA damaging agents in the supF assay are reviewed here, in the context of recent reports that relate to the nature of the DNA lesions responsible for the induced mutations. Mutations induced by nitric oxide in the supF assay are compared to those induced by N(2)O(3), nitrous acid, peroxynitrite and different reactive oxygen species. The effect of replication of the damaged pSP189 plasmid in human cells or Escherichia coli cells is also considered. PMID- 10838137 TI - Hydroperoxide-induced DNA damage and mutations. AB - Hydroperoxides (ROOH) are believed to play an important role in the generation of free radical damage in biology. Hydrogen peroxide (R=H) is produced by endogenous metabolic and catabolic processes in cells, while alkyl hydroperoxides (R=lipid, protein, DNA) are produced by free radical chain reactions involving molecular oxygen (autooxidation). The role of metal ions in generating DNA damage from hydroperoxides has long been recognized, and several distinct, biologically relevant mechanisms have been identified. Identification of the mechanistic pathways is important since it will largely determine the types of free radicals generated, which will largely determine the spectrum of DNA damage produced. Some mechanistic aspects of the reactions of low valent transition metal ions with ROOH and their role in mutagenesis are reviewed with a perspective on their possible role in the biological generation of DNA damage. A survey of hydroperoxide-induced mutagenesis studies is also presented. In vitro footprinting of DNA damage induced by hydroperoxides provides relevant information on sequence context dependent reactivity, and is valuable for the interpretation of mutation spectra since it represents the damage pattern prior to cellular repair. Efforts in this area are also reviewed. PMID- 10838138 TI - Mismatch repair deficient human cells: spontaneous and MNNG-induced mutational spectra in the HPRT gene. AB - We have determined both the spontaneous and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-induced mutational spectra in the HPRT gene of human cells (MT1) defective in the mismatch repair gene hMSH6 (GTBP). Eight of nine exons and nine of sixteen intronic flanking sequences were scanned, encompassing >900 bp of the HPRT gene. Mutant hotspots were detected and separated by differences in their melting temperatures using constant denaturant capillary electrophoresis (CDCE) or denaturing gradient gel electrophoresis (DGGE).A key finding of this work is that a high proportion of all HPRT inactivating mutations is represented by a small number of hotspots distributed over the exons and mRNA splice sites. Thirteen spontaneous hotspots and sixteen MNNG-induced hotspots accounted for 55% and 48% of all 6TG(R) point mutations, respectively. MNNG-induced hotspots were predominantly G:C-->A:T transitions. The spontaneous spectrum of cells deficient in hMSH6 contained transversions (A:T-->T:A, G:C-->T:A, A:T-->C:G), transitions (A:T-->G:C), a plus-one insertion, and a minus-one deletion. Curiously, G:C-->A:T transitions, which dominate human germinal and somatic point mutations were absent from the spontaneous hMSH6 spectra. PMID- 10838139 TI - Role of DNA repair in carcinogen-induced ras mutation. AB - In this contribution we discuss the gene- and cell type-specific repair of miscoding DNA alkylation products as a risk parameter in both mutation induction and malignant transformation by N-nitroso carcinogens. Upon exposure to N-nitroso compounds such as N-methyl-N-nitrosourea (MeNU) or N-ethyl-N-nitrosourea (EtNU), about a dozen different alkylation products are formed in cellular DNA. Among these are O(6)-methylguanine (O(6)-MeGua) and O(6)-ethylguanine (O(6)-EtGua), respectively, which differ only by one CH(2) group in their alkyl residue and, when unrepaired, cause G:C-->A:T transition mutations by anomalous base pairing during DNA replication. We have analyzed the global and gene-specific repair of O(6)-MeGua and O(6)-EtGua in target cell DNA, ras gene mutation frequencies, and tumor incidence, in the model of mammary carcinogenesis induced in 50-day-old female Sprague-Dawley rats by a single application of MeNU or EtNU. Both carcinogens induce histologically indistinguishable mammary adenocarcinomas at high yield. In the target mammary epithelia, O(6)-MeGua is repaired at similar slow rates in both transcriptionally active genes (Ha-ras, beta-actin), silent genes (lgE heavy chain), and in bulk DNA, by the one-step repair protein O(6) alkylguanine-DNA alkyltransferase (MGMT; low level of expression in the target cells). The slow repair of O(6)-MeGua translates into a high frequency of mutations at the central position of Ha-ras codon 12 (GGA) in MeNU-induced tumors. O(6)-EtGua, however, is removed approximately 20 times faster than O(6) MeGua selectively from transcribed genes via an MGMT independent, as yet uncharacterized excision mechanism. Accordingly, no Ha-ras codon 12 mutations are found in the EtNU-induced mammary tumors. Neither MeNU- nor EtNU-induced tumors exhibit mutations at codons 13 and 61 of Ha-ras or at codons 12, 13 and 61 of Ki ras. While a moderate surplus MGMT activity of the target cells - contributed by a bacterial MGMT transgene (ada) - significantly counteracts mammary tumorigenesis in MeNU-exposed rats, this is not the case in the EtNU-treated animals. Differential repair of structurally distinct DNA lesions in transcribed or (temporarily) silent genes thus determines the probability of mutation and, together with cell type-specific and interindividual differences in DNA repair capacity, influences carcinogenic risk. PMID- 10838140 TI - p53 mutational spectra and the role of methylated CpG sequences. AB - The occurrence of tumor-specific mutational spectra in the p53 mutation database provides indirect evidence that implicates certain exogenous and possibly endogenous mutagenic events in human carcinogenesis. In some cases, the distribution of DNA damage along the p53 gene caused by environmental carcinogens can be correlated with the mutational spectra, i.e. hotspots and types of mutations of certain cancers, most notably for nonmelanoma skin cancers and lung cancers in smokers. This concept has been validated by experiments with sunlight and cigarette smoke components representing the polycyclic aromatic hydrocarbon class of carcinogens. A disproportionally high number of mutations in p53 (and other genes) are found at methylated CpG dinucleotides. These sequences are particularly prone to mutagenesis involving endogenous events as well as modification by exogenous carcinogens. PMID- 10838141 TI - Mutagenesis and carcinogenesis in nucleotide excision repair-deficient XPA knock out mice. AB - Mice with a defect in the xeroderma pigmentosum group A (XPA) gene have a complete deficiency in nucleotide excision repair (NER). As such, these mice mimic the human XP phenotype in that they have a >1000-fold higher risk of developing UV-induced skin cancer. Besides being UV-sensitive, XPA(-/-) mice also develop internal tumors when they are exposed to chemical carcinogens. To investigate the effect of a total NER deficiency on the induction of gene mutations and tumor development, we crossed XPA(-/-) mice with transgenic lacZ/pUR288 mutation-indicator mice. The mice were treated with various agents and chemicals like UV-B, benzo[a]pyrene and 2-aceto-amino-fluorene. Gene mutation induction in several tumor target- and non-target tissues was determined in both the bacterial lacZ reporter gene and in the endogenous Hprt gene. Furthermore, alterations in the p53- and ras genes were determined in UV-induced skin tumors of XPA(-/-) mice. In this work, we review these results and discuss the applicability and reliability of enhanced gene mutant frequencies as early indicators of tumorigenesis. PMID- 10838142 TI - Studies of in vivo mutations in rpsL transgene in UVB-irradiated epidermis of XPA deficient mice. AB - We have established xeroderma pigmentosum group A (XPA) gene-knockout mice with nucleotide excision repair (NER) deficiency, which rapidly developed skin tumors when exposed to a low dose of chronic UV like XP-A patients, confirming that the NER process plays an important role in preventing UVB-induced skin cancer. To examine the in vivo mutation in the UVB-irradiated epidermis, we established XPA (-/-), (+/-) and (+/+) mice carrying the Escherichia coli rpsL transgene with which the mutation frequencies and spectra in the UVB-irradiated epidermal tissue can be examined conveniently. The XPA (-/-) mice showed a higher frequency of UVB induced mutation in the rpsL transgene with a low dose (150 J/m(2)) of UVB irradiation than the XPA (+/-) and (+/+) mice, while, at a high dose (900 J/m(2)) they showed almost the same frequency of mutation as the XPA (+/-) and (+/+) mice, probably because of cell death in the epidermis of the XPA (-/-) mice. However, CC-->TT tandem transition, a hallmark of UV-induced mutation, was detected at higher frequency in the XPA (-/-) mice than the XPA (+/-) and (+/+) mice at both doses of UVB. This rpsL/XPA mouse system will be useful for further analyzing the role of NER in the mutagenesis and carcinogenesis induced by various carcinogens. PMID- 10838143 TI - UV-specific mutations of the human patched gene in basal cell carcinomas from normal individuals and xeroderma pigmentosum patients. AB - Germline mutations of the human patched gene, PTCH, are responsible for the nevoid basal cell carcinoma (NBCC) syndrome or Gorlin's syndrome, characterized by multiple skin cancers, internal cancers and severe developmental abnormalities. The patched gene codes for a developmental regulator protein implicated in the sonic hedgehog (SHH) signalling pathway which plays an important role in oncogenic transformation. Patched exhibits tumor suppression function and has been shown to be mutated in skin cancers isolated from DNA repair-proficient patients or from xeroderma pigmentosum (XP), a DNA repair deficient syndrome. We have reviewed and analyzed in detail the different mutation spectra found on the PTCH gene in these various models. The type and distribution of mutations are quite different between germline, sporadic and XP cancers. Among the germline alterations, there is a preponderance (70%) of rearrangements compared to other tumour types analysed where less than 30% of rearrangements is observed. Typical UV-induced mutations of the patched gene are found prominently in XP basal cell carcinomas (BCCs) and in particular, a significantly higher level (63%) of the UV signature tandem mutations is found compared to sporadic BCC (11%). The location of mutations along the PTCH protein delineates several important functional domains implicated in the biology of this transmembrane receptor. PMID- 10838144 TI - Radiation specific patterns of loss of heterozygosity on chromosome 17q. AB - We and others have previously reported that the percentage of ionizing radiation induced TK(-) mutants exhibiting loss of heterozygosity (LOH) is not significantly different from those occurring spontaneously. In order to search further for a distinguishing feature of the X-ray-induced spectrum, and to characterize mechanisms of chromosomal scale mutagenesis, we used detailed mapping information to analyze the extent of LOH along chromosome 17q. Significant differences were observed when the extent of LOH tracts was considered. The representation of very long LOH tracts (>/=41 cM) was significantly (p=0.004) more common among spontaneous mutants, while relatively local LOH events, involving only markers in a 1-10 cM region surrounding the tk locus, are significantly (p=0.018) more prevalent among X-ray-induced mutants. Our data suggests that, although large deletions are recoverable, X-ray-induced autosomal deletions are not evenly distributed over the available size range. This indicates a mechanistic rather than biological restriction to the size of radiation-induced deletions, and demonstrates that the pattern of LOH may also be useful as a distinguishing component of the mutational spectrum. PMID- 10838145 TI - Hemispheric asymmetry in cortical control of memory-guided saccades. A transcranial magnetic stimulation study. AB - To study the temporal organisation of memory-guided saccade control we used single-pulse transcranial magnetic stimulation (TMS) over the left posterior parietal (PPC) and prefrontal cortex (PFC) in eight healthy subjects. TMS was applied either following presentation of a visual target, i.e. 160, 260, and 360 ms after the flashed point, or during the period of memorisation, i.e. between 700 and 1500 ms, or finally 100 ms after extinguishing of the central fixation point (i.e. 2100 ms after the target presentation). Latency of memory-guided saccades and the percentage of error in amplitude (PEA) was measured and compared with results without stimulation.TMS over the left PPC 100 ms after the extinguishing of the central fixation point significantly increased memory-guided saccade latency bilaterally. Furthermore, stimulation over the left PFC had a significant effect on the PEA of contralateral memory-guided saccades when applied during the period of memorisation, i.e. between 700 and 1500 ms.In a previous study using identical methodology [13: Muri RM, Vermersch SI, Rivaud S, Gaymard B, Pierrot-Deseilligny C. Effects of single-pulse transcranial magnetic stimulation over the prefrontal and posterior parietal cortices during memory guided saccades in humans. Journal of Neurophysiology 1996;76:2102-2106], we found that TMS over the right PPC increased the contralateral PEA when applied 260 ms after the flash, the effects on saccade latency after right PPC stimulation or on the PEA after right PFC stimulation being similar to those observed here. Taken together, these results show that (1) a hemispheric asymmetry in the preparation of memory-guided saccade amplitude during the early phase of sensorimotor integration exists, (2) memory-guided saccade triggering is controlled by PPC on both sides, and (3) PFC on both sides are involved in spatial working memory performance. PMID- 10838146 TI - Impaired planning but intact decision making in early Huntington's disease: implications for specific fronto-striatal pathology. AB - Previous neuropsychological data have suggested that deficits in early Huntington's disease (HD) include executive impairments, which often are linked with frontal-lobe dysfunction. This study sought to investigate the profile of cognitive deficits using two computerised tasks whose performance is known to rely on intact functions of separate areas of the prefrontal cortex. Twenty patients with early HD and 20 matched controls were given the one-touch Tower of London, a stringent measure of visuo-spatial planning, and a decision making task, which involved selecting and gambling on outcomes on the basis of their differing probabilities. Patients were significantly less accurate than controls on the planning test, which is sensitive to frontal lobe lesions and is strongly associated with the dorsolateral prefrontal cortex in functional imaging studies. On the decision making task, patients were unimpaired on the quality of their decision making, in contrast to previous reports of impairment on this task in patients with ventromedial prefrontal cortex lesions. This dissociation of performance is discussed in terms of the usual path of progression of HD through the striatum and the resultant pattern of disruption of the functioning of the different cortico-striatal functional loops. PMID- 10838147 TI - Alzheimer disease constricts the dynamic range of spatial attention in visual search. AB - A cued visual search task was used to examine the dynamic range over which spatial attention affects target identification during visual search. Precues varied in validity (valid, invalid, or neutral) and in precision (cue size) of target localization. Participants were "young-old" (65-74 years) and "old-old" (75-85 years) elderly adults and individuals in the mild stage of dementia of the Alzheimer type (DAT). For all participants, search was speeded as the precision with which a precue surrounding the location of a subsequently appearing target increased (precue size decreased). The cue size effect was evident in both feature and conjunction search, but was greatly reduced in both old-old and DAT groups compared to the young-old. However, whereas all non-demented adults showed a progressive modulation of search efficiency over the entire range of cue sizes, the dynamic range of spatial attention was restricted to the most precise cue in the DAT group. The restriction in the dynamic range of spatial attention may represent an underlying component of the impairment in perceptual and memory functioning found in early-stage DAT. PMID- 10838148 TI - Is the organisation of goal-directed action modality specific? A common temporal structure. AB - With the help of kinematic analysis, the temporal organization of the complex daily activity 'drinking from a bottle with a glass' was described in detail. The analysis focused on the sequential action structure, the prehensile acts, and the bimanual coordination as well as on the effect of different instruction modalities on these parameters to explore the underlying representation for this complex action. Movements of the two arms were recorded in three-dimensional space with the help of an optoelectronic device in 12 normal subjects under four conditions: (1) action pantomime after verbal instruction; (2) action imitation after observation of the action performed by the experimenter without the objects; (3) action pantomime while seeing, but not touching the objects; and finally (4) action execution with objects. Despite high execution variability, the temporal structure of the action could be precisely described by the relative duration and peak velocity of action segments, by the MGA-object size correlation, and by linear regression analysis between the onsets of functionally related action segments. A similar structure of the action as characterized by these kinematic parameters was retained across different instruction modalities. Only when the action was executed with the objects, the interval between the movement onsets of either hand and the peak velocity of the manipulative acts were reduced, while no change was observed across the other three instruction modalities. This stability of the temporal structure suggests the existence of a level in the representation of an action where all the modalities converge. PMID- 10838149 TI - Cortical activity during rotational and linear transformations. AB - Neuroimaging studies of cortical activation during image transformation tasks have shown that mental rotation may rely on similar brain regions as those underlying visual perceptual mechanisms. The V5 complex, which is specialised for visual motion, is one region that has been implicated. We used functional magnetic resonance imaging (fMRI) to investigate rotational and linear transformation of stimuli. Areas of significant brain activation were identified for each of the primary mental transformation tasks in contrast to its own perceptual reference task which was cognitively matched in all respects except for the variable of interest. Analysis of group data for perception of rotational and linear motion showed activation in areas corresponding to V5 as defined in earlier studies. Both rotational and linear mental transformations activated Brodman Area (BA) 19 but did not activate V5. An area within the inferior temporal gyrus, representing an inferior satellite area of V5, was activated by both the rotational perception and rotational transformation tasks, but showed no activation in response to linear motion perception or transformation. The findings demonstrate the extent to which neural substrates for image transformation and perception overlap and are distinct as well as revealing functional specialisation within perception and transformation processing systems. PMID- 10838150 TI - Vision and cognition in Alzheimer's disease. AB - Alzheimer's disease (AD) is known to affect visual pathways, but potential concomitant effects on vision and cognitive performance are not well understood. We studied 43 individuals with AD of mild severity and 22 individuals without dementia on a battery of tests designed to measure multiple aspects of basic and higher-order visual perception and cognition. All subjects performed on the same visual and cognitive test batteries. The results showed no differences between groups on tests of static visual acuity, stereoacuity, dynamic visual acuity or motion direction discrimination. However, individuals with AD performed significantly worse on tests of static spatial contrast sensitivity, visual attention, shape-from-motion, color, visuospatial construction and visual memory. Correlation analyses showed strong relationships between visual and cognitive scores. The findings show that AD affects several aspects of vision and are compatible with the hypothesis that visual dysfunction in AD may contribute to performance decrements in other cognitive domains. The pattern of involvement indicates that AD affects multiple visual neural pathways and regions. It is possible that better understanding of vision-related dysfunction could aid diagnosis and interventions to improve functional capacity in patients with dementia. PMID- 10838151 TI - A functional MRI case study of acquired cerebral dyschromatopsia. AB - Evidence from imaging studies suggests that primary visual cortex and multiple areas in ventral occipitotemporal cortex subserve color perception in humans. To learn more about the organization of these areas, we used structural and functional MRI (fMRI) to examine a patient with damage to ventral cortex. An art professor, KG, suffered a cerebrovascular accident during heart surgery that impaired his ability to perceive color. The Farnsworth-Munsell 100-Hue test was used to assess the extent of his deficit. When tested 12 months after the lesion, KG performed worse than 95% of age-matched normals on the 100-Hue test, but well above chance. Structural and functional MRI studies were conducted 3 years after the lesion to investigate the neuroanatomical correlates of KG'ss remaining color ability. Structural MRI revealed bilateral damage to ventral occipitotemporal cortex. In young and age-matched normal controls, an fMRI version of the 100-Hue reliably activated bilateral, color-selective regions in primary visual cortex and anterior and posterior ventral cortex. In subject KG, color-selective cortex was found in bilateral primary visual cortex. In ventral cortex, no color selective activity was observed in right ventral cortex, and only a small area of activity was observed in left anterior ventral cortex. However, significant color selective activity was observed in posterior left ventral cortex spared by the lesion. This posterior left ventral activation was similar in extent, position, and degree of color-selectivity to the posterior left posterior activation observed in normal controls, suggesting that this focus may be the cortical substrate underlying KG's remaining color perception. PMID- 10838152 TI - Drug abusers show impaired performance in a laboratory test of decision making. AB - A defining feature of drug addiction is persistent drug use despite long-term adverse consequences. This study examined the performance of drug abusers on a neuropsychological test that requires evaluation of long-term outcomes in the presence of a complex set of mixed reward/punishment contingencies (the Gambling Task). In order to control for generalized deficits related to choice and planning, subjects were also administered the Wisconsin Card Sorting Task. Thirty polysubstance abusers were compared to a comparison group of 24 subjects who did not use illicit drugs of abuse. Drug abusers performed much more poorly on the Gambling Task (net score = 10.2 +/- 4.7, mean +/- s.e.m.) than controls (26.0 +/- 5.3), but did not differ from controls on the Wisconsin Card Sorting Task. The results show that drug abusers are more likely to make maladaptive decisions in the Gambling Task that result in long-term losses exceeding short-term gains. These findings indicate that the Gambling Task may be a useful model in laboratory studies of cognitive dysfunctions associated with drug abuse. PMID- 10838153 TI - The effect of encoding manipulations on neural correlates of episodic retrieval. AB - The present experiments investigated whether the neural correlates of explicit (conscious) retrieval from episodic memory vary qualitatively according to conditions at encoding, as is predicted by current views of the neural basis of memory retrieval. Event-related potential (ERP) correlates of word stem (e.g. MOT_) cued recall were compared for items studied under different encoding conditions. In Experiment 1, encoding was either 'shallow' or 'deep' whereas in Experiment 2 the presentation modality of the study items was either visual or auditory. In both experiments robust ERP memory effects were observed for stems completed with explicitly retrieved items from each encoding condition. The effects varied in their magnitude, such that they were largest when elicited by the more memorable class of item in each experiment. The scalp distributions of the effects did not differ according to encoding condition, a finding which offers no support for the view that retrieval involves the literal reinstatement of neural activity engaged at the time of encoding. The findings instead point to the existence of a set of retrieval operations that are engaged regardless of the conditions under which retrieved information is encoded. PMID- 10838154 TI - Endothelial cell responses to hypoxia: initiation of a cascade of cellular interactions. AB - The origin of several vascular pathologies involves sudden or recurrent oxygen deficiency. In this review, we examine what the biochemical and molecular responses of the endothelial cells to the lack of oxygen are and how these responses may account for the features observed in pathological situations, mainly by modifications of cell-cell interactions. Two major responses of the endothelial cells have been observed depending on the degree and duration of the oxygen deficiency. Firstly, acute hypoxia rapidly activates the endothelial cells to release inflammatory mediators and growth factors. These inflammatory mediators are able to recruit and promote the adherence of neutrophils to the endothelium where they become activated. The synthesis of platelet-activating factor plays a key role in this adherence process. Secondly, longer periods of hypoxia increase the expression of specific genes such as those encoding some cytokines as well as for the growth factors platelet-derived growth factor and vascular endothelial growth factor. The transcriptional induction of these genes is mediated through the activation of several transcription factors, the most important one being hypoxia inducible factor-1. The link between our knowledge of the signalling cascade of the cellular and molecular events initiated by hypoxia and their involvement in several vascular pathological situations, varicose veins, tumor angiogenesis and pulmonary hypertension is discussed briefly. PMID- 10838155 TI - Store-operated Ca(2+) inflow in Reuber hepatoma cells is inhibited by voltage operated Ca(2+) channel antagonists and, in contrast to freshly isolated hepatocytes, does not require a pertussis toxin-sensitive trimeric GTP-binding protein. AB - The treatment of H4-IIE cells (an immortalised liver cell line derived from the Reuber rat hepatoma) with thapsigargin, 2, 5-di-(tert-butyl)-1,4 benzohydroquinone, cyclopiazonic acid, or pretreatment with EGTA, stimulated Ca(2+) inflow (assayed using intracellular fluo-3 and a Ca(2+) add-back protocol). No stimulation of Mn(2+) inflow by thapsigargin was detected. Thapsigargin-stimulated Ca(2+) inflow was inhibited by Gd(3+) (maximal inhibition at 2 microM Gd(3+)), the imidazole derivative SK&F 96365, and by relatively high concentrations of the voltage-operated Ca(2+) channel antagonists, verapamil, nifedipine, nicardipine and the novel dihydropyridine analogues AN406 and AN1043. The calmodulin antagonists W7, W13 and calmidazolium also inhibited thapsigargin induced Ca(2+) inflow and release of Ca(2+) from intracellular stores. No inhibition of either Ca(2+) inflow or Ca(2+) release was observed with calmodulin antagonist KN62. Substantial inhibition of Ca(2+) inflow by calmidazolium was only observed when the inhibitor was added before thapsigargin. Pretreatment of H4-IIE cells with pertussis toxin, or treatment with brefeldin A, did not inhibit thapsigargin-stimulated Ca(2+) inflow. Compared with freshly isolated rat hepatocytes, H4-IIE cells exhibited a more diffuse actin cytoskeleton, and a more granular arrangement of the endoplasmic reticulum (ER). In contrast to freshly isolated hepatocytes, the arrangement of the ER in H4-IIE cells was not affected by pertussis toxin treatment. Western blot analysis of lysates of freshly isolated rat hepatocytes revealed two forms of G(i2(alpha)) with apparent molecular weights of 41 and 43 kDa. Analysis of H4-IIE cell lysates showed only the 41 kDa form of G(i2(alpha)) and substantially less total G(i2(alpha)) than that present in rat hepatocytes. It is concluded that H4-IIE cells possess store operated Ca(2+) channels which do not require calmodulin for activation and exhibit properties similar to those in freshly isolated rat hepatocytes, including susceptibility to inhibition by relatively high concentrations of voltage-operated Ca(2+) channel antagonists. In contrast to rat hepatocytes, SOCs in H4-IIE cells do not require G(i2(alpha)) for activation. Possible explanations for differences in the requirement for G(i2(alpha)) in the activation of Ca(2+) inflow are briefly discussed. PMID- 10838156 TI - Differential activation and redistribution of c-Src and Fyn in platelets, assessed by MoAb specific for C-terminal tyrosine-dephosphorylated c-Src and Fyn. AB - Tyrosine kinases, c-Src and Fyn, in their active form, have their C-terminal tyrosine residue dephosphorylated. In this study, we used clone 28, a monoclonal antibody (MoAb) that recognizes dephosphorylated C-terminal tyrosine of c-Src and Fyn, to investigate the mode of activation and mobilization of these kinases. Independently of integrin alphaIIbbeta3 signaling, the Fyn activity increased by 8.3-fold 5 s after stimulation with 20 microM TRAP (thrombin receptor agonist peptide), while that of c-Src increased only by 2.9-fold 15 s after stimulation. Both c-Src and Fyn translocated to the Triton-insoluble cytoskeletal fraction in an aggregation-dependent manner. Five minutes after TRAP-stimulation, 85% of Fyn translocated to the cytoskeleton, while only about 20% of c-Src was recovered in this fraction. The Triton-insoluble fraction was further fractionated by RIPA (radioimmunoprecipitation assay) buffer containing 0.1% SDS. While active c-Src was predominantly present in the Triton-insoluble/RIPA-insoluble fraction, clone 28-negative c-Src was present in the Triton-insoluble/RIPA-soluble fraction. On the other hand, Fyn was present only in the Triton-insoluble/RIPA-insoluble fraction. These findings suggest that the mode of activation and redistribution into the cytoskeleton differs between c-Src and Fyn, and that clone 28 provides a useful tool for investigating the activation and mobilization of Src family tyrosine kinases. PMID- 10838157 TI - Release from apoptosis correlates with tumor progression in the AKR lymphoma. AB - Disturbance of apoptosis is an established factor in tumorigenesis. The role of apoptosis in tumor progression is not yet clear. In the present study we compared the tendency to spontaneous apoptosis (and the proliferative capacity) of tumor cells derived from primary (PT) and metastatic tumor (MT) cells of several AKR lymphoma variants. Apoptosis-related gene expression was also compared. Our results indicate that release from apoptosis has a role in the tumor progression of this T cell lymphoma. At the cellular level, a markedly lower apoptotic tendency was observed in MT than in PT cells. The existence of macrophages only in PT also supports the presence of apoptotic cells in local but not in MTs. By contrast, proliferative capacity does not determine tumor aggressiveness in this system. At the molecular level, we found a higher staining intensity for bcl-2 in MT than in PT cells, suggesting that bcl-2 might be responsible for the reduced apoptosis in MT compared to PT cells. Evidence for p53 overexpression was found in the MT cells of one of the variants but in none of the PT. Comparison of Fas receptor, unexpectedly showed an increased expression in MT versus PT cells, possibly indicating resistance to Fas-induced apoptosis in the MT cells. PMID- 10838158 TI - The effects of Tempol on ferritin synthesis and Fe metabolism in lens epithelial cells. AB - The nitroxide, Tempol, can protect tissue from oxidative damage by removing superoxide and by oxidizing Fe(II) to Fe(III), thus decreasing formation of the hydroxyl radical. However, long-term exposure to Tempol can damage cells. The oxidation of Fe could have profound effects on Fe metabolism in cells, yet this has not been previously studied. In the present investigation, the effects of Tempol on the synthesis of the Fe storage protein, ferritin, and its ability to store Fe were studied in cultured lens epithelial cells (LEC). In addition, the effects of short- and long-term Tempol treatment on the resistance of LEC to oxidative stress were determined. Tempol had a clear dose-dependent inhibitory effect on ferritin synthesis noted at 6 h. By 20 h, ferritin synthesis returned toward normal levels. However, Fe incorporation into ferritin was decreased by almost 90% by the highest dose of Tempol, even at the 20-h time point. The decrease in Fe incorporation into ferritin was accompanied by a significant increase in the LMW pool of Fe. After short-term (4 h) treatment with Tempol, LEC were protected against the toxic effects of tertiary butyl hydroperoxide. However, after longer term treatment (20 h), Tempol itself had a toxic effect and did not afford protection. Indeed, at the higher doses, Tempol significantly reduced the ability of the cells to withstand oxidative stress. The redistribution of Fe within the cell after 20 h of Tempol treatment appears to render the cells more vulnerable to oxidative stress. The deleterious effects of Tempol on LEC are likely due to its effects on Fe metabolism, perhaps by reducing the availability of Fe for incorporation into ferritin and Fe-dependent enzymes as well as enlarging a low molecular weight pool of Fe which may be capable of catalyzing damaging free radical reactions. PMID- 10838159 TI - Dexamethasone inhibits the induction of NAD(+)-dependent 15-hydroxyprostaglandin dehydrogenase by phorbol ester in human promonocytic U937 cells. AB - Pro-inflammatory prostaglandins are known to be first catabolized by NAD(+) dependent 15-hydroxyprostaglandin dehydrogenase (15-PGDH) to inactive metabolites. This enzyme is under regulatory control by various inflammation related agents. Regulation of this enzyme was investigated in human promonocytic U937 cells. 15-PGDH activity was found to be optimally induced by phorbol 12 myristate 13-acetate (PMA) at 10 nM after 24 h of treatment. The induction was blocked by staurosporine or GF 109203X indicating that the induction was mediated by protein kinase C. The induction by PMA was inhibited by the concurrent addition of dexamethasone. Nearly complete inhibition was observed at 50 nM. Other glucocorticoids, such as hydrocortisone and corticosterone, but not sex hormones, were also inhibitory. Inhibition by dexamethasone could be reversed by the concurrent addition of antagonist mifepristone (RU-486) indicating that the inhibition was a receptor-mediated event. Either induction by PMA or inhibition by dexamethasone the 15-PGDH activity correlated well with the enzyme protein expression as shown by the Western blot analysis. These results provide the first evidence that prostaglandin catabolism is regulated by glucocorticoids at the therapeutic level. PMID- 10838160 TI - Mechanism of transforming growth factor-beta1-induced expression of vascular endothelial growth factor in murine osteoblastic MC3T3-E1 cells. AB - Transforming growth factor-beta1 (TGF-beta1), an abundant growth factor in bone matrix, has been shown to be involved in bone formation and fracture healing. The mechanism of action of the osteogenic effect of TGF-beta1 is not clearly understood. In this study, we found that the addition of TGF-beta1 to murine osteoblastic MC3T3-E1 cells induced vascular endothelial growth factor (VEGF) mRNA production. VEGF mRNA levels reached a plateau within 2 h after the addition of TGF-beta1. The induction was superinduced by cycloheximide and blocked by actinomycin D. Ro 31-8220, a protein kinase C inhibitor, abrogated the induction. In addition, curcumin, an inhibitor for transcription factor AP-1, also blocked the induction. Electrophoretic mobility shift assay revealed an enhanced binding of transcription factors AP-1 and NF-kappaB. Transient transfection experiment showed that VEGF promoter activity increased 3.6-fold upon TGF-beta1 stimulation. Immunoblot analysis showed that the amount of secreted VEGF was elevated in the medium 4 h after TGF-beta1 stimulation. Our results therefore suggest that at least part of the osteogenic activity of TGF-beta1 may be attributed to the production of VEGF. PMID- 10838161 TI - Differential adaptation of growth and differentiation factor 8/myostatin, fibroblast growth factor 6 and leukemia inhibitory factor in overloaded, regenerating and denervated rat muscles. AB - Mice genetically deficient in growth and differentiation factor 8 (GDF8/myostatin) had markedly increased muscle fiber numbers and fiber hypertrophy. In the regenerating muscle of mice possessing FGF6 mutation, fiber remodeling was delayed. Although myostatin and FGF6 may be important for the maintenance, regeneration and/or hypertrophy of muscle, little work has been done on the possible role of these proteins in adult muscle in vivo. Using Western blot and immunohistochemical analysis, we investigated, in rats, the distribution of myostatin, FGF6 and LIF proteins between slow- and fast-type muscles, and the adaptive response of these proteins in mechanically overloaded muscles, in regenerating muscles following bupivacaine injection and in denervated muscles after section of the sciatic nerve. The amounts of myostatin and LIF protein were markedly greater in normal slow-type muscles. In the soleus muscle, myostatin and LIF proteins were detected at the site of the myonucleus in both slow-twitch and fast-twitch fibers. In contrast, FGF6 protein was selectively expressed in normal fast-type muscles. Mechanical overloading rapidly enhanced the myostatin and LIF but not FGF6 protein level. In the regenerating muscles, marked diminution of myostatin and FGF6 was observed besides enhancement of LIF. Denervation of fast type muscles rapidly increased the LIF, but decreased the FGF6 expression. Therefore, the increased expressions of myostatin and LIF play an important role in muscle hypertrophy following mechanical overloading. The marked reduction of FGF6 in the hypertrophied and regenerating muscle would imply that FGF6 regulates muscle differentiation but not proliferation of satellite cells and/or myoblasts. PMID- 10838162 TI - Prolactin stimulation of tyrosyl phosphorylation of Shc proteins in Nb(2) lymphoma cells, but not mammary tissues. AB - Prolactin (PRL) stimulates lactogenesis in mammary cells and mitogenesis in a variety of cell types including Nb(2) cells. Studies indicate that a different composite of signaling pathways is involved in the PRL stimulation of mitogenesis as compared to lactogenesis. In the present studies, PRL is shown to stimulate the tyrosyl phosphorylation of all three isoforms of Shc proteins in Nb(2) cells (mitogenesis), but not in the mammary gland. Maximal phosphorylation of the Shc proteins is expressed between 10 and 15 min after a 50-ng/ml PRL treatment. In addition, there is an increased association between the Grb2 protein and Shc proteins upon PRL stimulation. However, no increased association between JAK2 and Shc proteins was observed in either the Nb(2) cells or mammary tissues. PMID- 10838163 TI - Molecular cloning of a ferret angiotensin II AT(1) receptor reveals the importance of position 163 for Losartan binding. AB - A complementary DNA for the angiotensin II (AngII) type 1 (AT(1)) receptor from Mustela putorius furo (ferret) was isolated from a ferret atria cDNA library. The cDNA encodes a protein (fAT(1)) of 359 amino acids having high homologies (93 99%) to other mammalian AT(1) receptor counterparts. When fAT(1) was expressed in COS-7 cells and photoaffinity labeled with the photoactive analogue (125)I ?Sar(1), Bpa(8)AngII, a protein of 100 kDa was detected by autoradiography. The formation of this complex was specific since it was abolished in the presence of the AT(1) non-peptidic antagonist L-158,809. Functional analysis indicated that the fAT(1) receptor efficiently coupled to phospholipase C as demonstrated by an increase in inositol phosphate production following stimulation with AngII. Binding studies revealed that the fAT(1) receptor had a high affinity for the peptide antagonist ?Sar(1), Ile(8)AngII (K(d) of 5. 8+/-1.4 nM) but a low affinity for the AT(1) selective non-peptidic antagonist DuP 753 (K(d) of 91+/ 15.6 nM). Interestingly, when we substituted Thr(163) with an Ala residue, which occupies this position in many mammalian AT(1) receptors, we restored the high affinity of this receptor for Dup 753 (11.7+/-5.13 nM). These results suggest that position 163 of the AT(1) receptor does not contribute to the overall binding of peptidic ligands but that certain non-peptidic antagonists such as Dup 753 are clearly dependent on this position for efficient binding. PMID- 10838164 TI - Regulation of human PLD1 and PLD2 by calcium and protein kinase C. AB - Numerous studies show that PLD is activated in cells by calcium and by protein kinase C (PKC). We found that human PLD1 and PLD2 expressed in Sf9 cells can be activated by calcium-mobilizing agonists and by co-expression with PKCalpha. The calcium-mobilizing agonists A23187 and CryIC toxin triggered large increases in phosphatidylethanol (PtdEth) production in Sf9 cells over-expressing PLD1 and PLD2, but not in vector controls. PLD activation by these agonists was largely dependent on extracellular calcium. Membrane assays demonstrated significant PLD1 and PLD2 activity in the absence of divalent cations, which could be enhanced by low levels of calcium either in the presence or absence of magnesium. PLD1 but not PLD2 activity was slightly enhanced by magnesium. Treatment of Sf9 cells expressing PLD1 and PLD2 with PMA resulted in little PtdEth production. However, a significant and comparable formation of PtdEth occurred when PLD1 or PLD2 were co-expressed with PKCalpha, but not PKCdelta, and was further augmented by PMA. In contrast to PLD1, co-expressing PLD2 with PKCalpha or PKCdelta further enhanced A23187-induced PtdEth production. Immunoprecipitation experiments demonstrated that PLD1 and PLD2 associated with the PKC isoforms in Sf9 cells. Furthermore, in membrane reconstitution assays, both PLD1 and PLD2 could be stimulated by calmodulin and PKCalpha-enriched cytosol. The results indicate that PLD2 as well as PLD1 is subject to agonist-induced activation in intact cells and can be regulated by calcium and PKC. PMID- 10838165 TI - Xenobiotic-mediated production of superoxide by primary cultures of rat cerebral endothelial cells, astrocytes, and neurones. AB - Previous works of our group demonstrated that xenobiotic metabolism by brain microsomes or cultured cerebral cells may promote the formation of reactive oxygen species. In order to characterise the risk of oxidative stress to both the central nervous system and the blood-brain barrier, we measured in the present work the release of superoxide in the culture medium of rat cerebrovascular endothelial cells during the metabolism of menadione, anthraquinone, diquat or nitrofurazone. Assays were run in the same experimental conditions on primary cultures of rat neurones and astrocytes. Quinone metabolism efficiently produced superoxide, but the production of radicals during the metabolism of diquat or nitrofurazone was very low, as a probable result of their reduced transport inside the cells. In all cell types assayed, superoxide production was time- and concentration-dependent, and cultured astrocytes always produced the highest amounts of radicals. Superoxide formation by microsomes prepared from the cultured cells was decreased by immunoinhibition of NADPH-cytochrome P450 reductase or by its irreversible inhibition by diphenyliodonium chloride, suggesting the involvement of this flavoprotein in radical production. Cerebrovascular endothelial cells cultured on collagen-coated filters produced equivalent amounts of superoxide both at their luminal side and through the artificial basement membrane, suggesting that in vivo, endothelial superoxide production may endanger adjacent astrocytes and neurones. PMID- 10838166 TI - Oversulfated fucoidan inhibits the basic fibroblast growth factor-induced tube formation by human umbilical vein endothelial cells: its possible mechanism of action. AB - We have previously demonstrated that chemically oversulfated fucoidan (OSF) but not native fucoidan (NF) effectively suppresses the tube structure formation by human umbilical vein endothelial cells (HUVEC) on the basement membrane preparation, Matrigel. In this study, using more defined systems where basic fibroblast growth factor (bFGF) induces the tube formation by HUVEC on collagen gel, we investigated the mechanism responsible for the inhibition of angiogenesis by OSF in vitro. Unlike NF and desulfated fucoidan (desF), OSF potently inhibited the bFGF-induced HUVEC migration and tube formation. ELISA for tissue-type plasminogen activator (t-PA) and plasminogen activator inhibitor-1 (PAI-1) in the culture media indicated that OSF increased the bFGF-induced release of PAI-1 antigen, but not of t-PA antigen. Analyses of the binding of bFGF to HUVEC surfaces and the following protein tyrosine phosphorylation revealed that OSF could promote the cell binding and autophosphorylation of 140 and 160 kDa receptors. In heparitinase-treated HUVEC, contrarily, the bFGF binding and PAI-1 release were decreased by OSF. These results suggest that OSF is a highly sulfated unique polysaccharide that can promote the binding of bFGF to the heparan sulfate molecules required for binding to the high affinity receptors with tyrosine kinase activity. The resultant increase in PAI-1 release may play a key role for the prevention of cell migration accompanied by matrix proteolysis. PMID- 10838167 TI - Cell cycle-independent induction of D1 and D2 cyclin expression, but not cyclin Cdk complex formation or Rb phosphorylation, by IFNgamma in macrophages. AB - During G1 to S phase transition, D-type cyclins form complexes with cyclin dependent kinases (Cdk), which in turn phosphorylate retinoblastoma gene product (Rb) and inhibit its growth-inhibitory function, leading ultimately to cell proliferation. We report here a novel finding that D1 and D2 cyclins are induced in macrophages by antiproliferative factor gamma interferon (IFNgamma). The induction appears to be transcriptional activation of the D cyclin genes, since indirect events such as IFNgamma-induced colony-stimulating factor-1 (CSF-1) autocrine stimulation, alteration of D1 and D2 mRNA stability and lipopolysaccharide contamination in commercial IFNgamma preparations play no roles. In contrast to CSF-1, IFNgamma neither induces D1-Cdk4 complex formation and Rb hyperphosphorylation nor interferes with CSF-1-stimulated D1-Cdk4 interaction and Rb phosphorylation, while it completely blocks CSF-1-stimulated cell proliferation. This study suggests that induction of D1 and D2 cyclins is not necessarily associated with cell cycle progression, and D cyclins may have cell cycle-independent functions in response to IFNgamma. PMID- 10838168 TI - Peroxisome proliferator-activated receptor gamma1 (PPARgamma1) expresses in rat mesangial cells and PPARgamma agonists modulate its differentiation. AB - Thiazolidinediones, synthetic ligands of peroxisome proliferator-activated receptor gamma (PPARgamma), are reported to have direct beneficial effects on diabetic nephropathy without lowering blood glucose levels in human and rat. We hypothesized these effects of thiazolidinediones might be derived from PPARgamma activation of kidney cells, and we examined the expression of PPARgamma and the effect of PPARgamma agonists, troglitazone and 15-deoxy-delta-prostaglandin J2 (15d-PGJ2), on the proliferation and differentiation in rat mesangial cells. A single band of mRNA of PPARgamma with a predicted size was detected in reverse transcription-polymerase chain reaction products (RT-PCR) using established PCR probes of PPARgamma. PPARgamma protein in rat mesangial cells was identified as PPARgamma1 by a Western blot. In a gel mobility shift assay to determine a binding activity of PPARgamma, the nuclear protein from rat mesangial cells bound to a (32)P-labeled oligonucleotide probe, including PPAR response elements. A synthetic and a natural ligand of PPARgamma, troglitazone and 15d-PGJ2, decreased thymidine incorporation in a dose dependent manner. After 7 days incubation with troglitazone and 15d-PGJ2, alpha-smooth muscle actin expression, a marker of mesangial cell de-differentiation, was decreased significantly compared to that of control. These results indicate that PPARgamma1 is expressing in rat mesangial cells, and PPARgamma1 activation with its agonists modulates the proliferation and differentiation of cultured rat mesangial cells. PMID- 10838169 TI - A possible role for Ca(2+)/calmodulin-dependent protein kinase IV during pancreatic acinar stimulus-secretion coupling. AB - Ca(2+)/calmodulin-dependent protein kinases (CaMKs) are important intracellular mediators in the mediation of stimulus-secretion coupling and excitation contraction coupling in a wide variety of cell types. We attempted to identify and characterize the functional roles of CaMK in mediating pancreatic enzyme secretion. Immunoprecipitation and immunoblotting studies using a CaMKII or CaMKIV antibody showed that rat pancreatic acini expressed both CaMKII and CaMKIV. Phosphotransferase activities of CaMKs were measured by a radioenzyme assay (REA) using autocamtide II, peptide gamma and myosin P-light chain as substrates. Although CaMKII and CaMKIV use autocamtide II as a substrate, peptide gamma is more efficiently phosphorylated by CaMKIV than by CaMKII. Intact acini were stimulated with cholecystokinin (CCK)-8, carbachol (CCh) and the high affinity CCK-A receptor agonist, CCK-OPE, and the cell lysates were used for REA. CCK-8, CCh and CCK-OPE caused a concentration-dependent increase in CaMKs activities. When autocamtide II was used, maximal increases were 1.5-1.8-fold over basal (20.2+/-2.0 pmol/min/mg protein), with peaks occurring at 20 min after cell stimulation. In separate studies that used peptide gamma, CCK-8, CCh and CCK OPE dose-dependently increased CaMKIV activities. Maximal increases were 1.5-2.4 fold over basal (30.7+/-3. 2 pmol/min/mg protein) with peaks occurring at 20 min after cell stimulation. Peak increases after cell stimulation induced by peptide gamma were 1.8-2.8-fold higher than those induced by autocamtide II. CCK-8, CCh and CCK-OPE also significantly increased phosphotransferase activities of myosin light chain kinase (MLCK) substrate (basal: 4.4+/-0.7 pmol/min/mg protein). However, maximal increases induced by MLCK substrate were less than 10% of those occurring in peptide gamma. Characteristics of the phosphotransferase activity were also different between autocamtide II and peptide gamma. When autocamtide II was used, elimination of medium Ca(2+) in either cell lysates or intact cells resulted in a significant decrease in the activity, whereas it had no or little effect when peptide gamma was used. This suggests that Ca(2+) influx from the extracellular space is not fully required for CaMKIV activity and Ca(2+) is not a prerequisite for phosphotransferase activity once CaMKIV is activated by either intracellular Ca(2+) release or intracellular Ca(2+) oscillations. The specific CaMKII inhibitor KN-62 (50 microM) had no effect on the CaMKIV activity and pancreatic enzyme secretion elicited by CCK-8, CCh and CCK-OPE. The specific MLCK inhibitor, ML-9 (10 microM), also did not inhibit CCK-8-stimulated pancreatic amylase secretion. In contrast, wide spectrum CaMK inhibitors, K-252a (1 microM) and KT5926 (3 microM), significantly inhibited CaMKIV activities and enzyme secretion evoked by secretagogues. Thus, CaMKIV appears to be an important intracellular mediator during stimulus-secretion coupling of rat pancreatic acinar cells. PMID- 10838170 TI - Cloning and expression studies during vegetative and sexual development of Pbs1, a septin gene homologue from Pyrenopeziza brassicae. AB - A septin gene homologue designated Pyrenopeziza brassicae septin 1 (Pbs1) has been identified and cloned from the plant pathogenic fungus Pyrenopeziza brassicae and its expression analysed. Pbs1 is present in both mating types and in a single copy within each genome and is transcribed in proportionate levels during both vegetative and sexual growth. PMID- 10838171 TI - Plant stress-related uncoupling protein CSP 310 caused lipid peroxidation in winter wheat mitochondria under chilling stress. AB - The effect of CSP 310 on lipid peroxidation in winter wheat mitochondria was studied by the measurement of primary lipid peroxidation products - dienic conjugates. It was found that some concentrations of CSP 310 caused lipid peroxidation in isolated winter wheat mitochondria in all systems investigated at different concentrations during chilling stress. PMID- 10838172 TI - Extracting operative temperatures from temperatures of physical models with thermal inertia. AB - Temperatures of operative temperature models, particularly those of thick-walled models of larger ectotherms, lag behind and are more restricted in range than the operative temperatures they estimate.Algorithms are provided to extract estimates of instantaneous operative temperatures from model temperatures.A simple deconvolution method can be used when wind speeds are constant.An iterative estimation method must be used when wind speed varies during the monitoring period.The iterative method is sensitive to measurement error, and so uses a smoothing filter to limit instabilities. The smoothing also limits the short-term fluctuations in the estimated operative temperature.Iterative estimates of operative temperature suggested time lags of up to 90 min between predicted operative temperatures and model temperatures for desert tortoises (mass=3 kg). Differences this large could affect estimates of time available for foraging. PMID- 10838173 TI - Cold tolerance in Drosophila: adaptive variations revealed by the analysis of starvation survival reaction norms. AB - Three species of Drosophila were investigated for their capacity to survive without food (starvation tolerance) at seven different temperatures ranging from 0 to 25 degrees C. In all cases biphasic response curves (reaction norms) were observed, corresponding either to special deleterious effects of cold or to a progressive exhaustion of reserves proportional to metabolic rate. The temperature at which survival was longest was called the threshold temperature. The position of the threshold exhibited adaptive changes, either due to acclimation in the same species, or to genetic variations evidenced between species. In D. melanogaster, adults grown at lower temperature (12 degrees C) were more tolerant to cold than adults grown at higher temperatures (21, 25 or 30 degrees C). This acclimation process shifted, in an adaptive way, the position of the threshold temperature from 6.2 to 7.5 degrees C. A comparison of three different species grown at a single developmental temperature (21 degrees C) revealed similar but greater adaptive differences in their threshold temperature: 4.8 degrees C in the temperate D. subobscura, 7 degrees C in the cosmopolitan D. melanogaster and 14.6 degrees C in the tropical D. ananassae. PMID- 10838174 TI - Sorbitol accumulation in whiteflies: evidence for a role in protecting proteins during heat stress. PMID- 10838175 TI - Heat shock proteins in whiteflies, an insect that accumulates sorbitol in response to heat stress. PMID- 10838176 TI - Cardiac tissue function of the teleost fish Oreochromis niloticus under different thermal conditions. AB - The cardiac responses of Oreochromis niloticus acclimated to 25 degrees C were assessed using ventricle strips mounted for isometric force recording (Fc) and in vivo heart rate (f(H)). f(H) increased progressively from 25 to 40 degrees C. At extracellular Ca(2+) concentrations of 1.25 and 9.25 mM, a transition from 25 to 40 degrees C resulted in a decreased Fc. At both 25 and 40 degrees C, Fc rose when [Ca(2+)] was increased from 1.25 to 9.25 mM. Fc remained constant at 72 and 120 contractions.min(-1) at 25 and 40 degrees C, respectively, and declined thereafter. The post-rest potentiation was not influenced by ryanodine, indicating that the sarcoplasmic reticulum is not important to the excitation contraction coupling. PMID- 10838177 TI - Hyperthermia-induced changes in the vascular permeability of rats: a model system to examine therapeutic interventions. AB - Extravasation in the heart, liver, lung, kidney, spleen, gastrocnemius, and duodenum was quantified in normothermic and hyperthermic (core temperature (T(c))=41.5, 42, or 42.6 degrees C) rats. Following attainment of the target T(c), Evans blue (Eb) was administered via jugular cannula; the animals were anesthetized, exsanguinated, tissues removed and washed in saline, and Eb extracted with formamide. There was significantly (p<0.05) more Eb (ug/g of dry wt of tissue, mean+/-SD) in the tissues of severely hyperthermic (T(c)=42.6 degrees C) rats vs that of control rats: liver - 198+/-39 vs 125+/-28, kidney - 376+/-68 vs 176+/-60, and small intestine - 170+/-49 vs 106+/-20. This model may be useful in evaluating the efficacy of treatment modalities designed to sustain vascular integrity in the face of environmental insult. PMID- 10838178 TI - Hyperthermic effects on reticuloendothelial system particulate uptake. AB - Reticuloendothelial system (RES) particulate uptake (PU) of vascular debris influences survival from extreme hyperthermia. Little is known of the effect of extreme hyperthermia, unrelated to fever, on RES PU shortly after reaching a maximum core temperature (T(c)). Relative to normothermic rats (T(c)=38.0 degrees C), rats at T(c)=42.6 degrees C had significantly higher, while T(c)=42.0 degrees C rats had significantly lower total RES tissue (lung, liver, spleen) PU of fluorescent microspheres (1 u), when compared to rats at T(c)=42.6 or 38.0 degrees C. These findings suggest at T(c)=42.6 degrees C, rats were not actively thermoregulating. As such, more blood remained in the core than in the periphery, which resulted in greater core RES tissue PU. In contrast, to reduce or control core heat, rats at T(c)=42.0 degrees or 38.0 degrees C directed more blood to the periphery, which reduced core RES tissue PU. Blood flow patterns as directed by the state or degree of active thermoregulation is likely an influence of hyperthermia on RES PU. PMID- 10838179 TI - Body temperatures of house mice artificially selected for high voluntary wheel running behavior: repeatability and effect of genetic selection. AB - We studied rectal body temperatures of house mice (Mus domesticus) that had been artificially selected for high voluntary wheel running.1. At generation 17, mice from the four replicate selected lines ran, on average, 2.5-times as many revolutions/day as did mice from the four random-bred control lines.2. During the day, repeatability of individual differences in body temperature measured 4 days apart was low; at night, repeatability was statistically significant across three time scales (1 day, 1 week, 2 weeks).3. During the day, body temperatures of selected and control animals did not differ; at night, mice from selected lines had higher body temperatures. However, when amount of wheel running immediately prior to measurement was included as a covariate, the difference was no longer statistically significant.Higher body temperatures, associated with increased activity, might enhance locomotor abilities through Q10 effects, increase metabolic rate and food requirements, affect sleep patterns, and alter expression of heat-shock proteins. PMID- 10838180 TI - Immune response in cold exposures of different types. PMID- 10838181 TI - Expression of five iduronate-2-sulfatase site-directed mutations. AB - Five point mutations (R88H, R88P, T118I, 959delT, R468Q) previously identified in the iduronate-2-sulfatase (IDS) gene of Italian Hunter patients were expressed in COS cells to evaluate their functional consequence on enzyme activity, processing and intracellular localization. The 88 arginine residue belongs to the CXPSR pentapeptide conserved in all human sulfatases, where cysteine modification to formylglycine is required for enzyme activity. Substitution of arginine with histidine residue resulted in 13.7% residual enzyme activity, with an apparent K(m) value (133 microM) lower than that found for the normal enzyme (327 microM), indicating a higher affinity for the substrate; substitution of arginine with proline resulted in total absence of residual activity, in agreement with the phenotypes observed in patients carrying R88H and R88P mutations. For the four missense mutations, pulse-chase labelling experiments showed an apparently normal maturation; however, subcellular fractionation demonstrated poor transport to lysosomes. Therefore, residues 88, 118 and 468 appear to be not essential for processing but important for IDS conformation. PMID- 10838182 TI - Detection of antibody to sialyl-i, a possible antigen in patients with Meniere's disease. AB - An autoimmune hypothesis for the etiology of Meniere's disease has been proposed. In this study, we focused on gangliosides as potential antigens for autoantibodies in Meniere's disease patients. In an attempt to investigate ganglioside antigens which respond to the serum of patients with Meniere's disease, we analyzed gangliosides of human acoustic neurinomas, and used them as antigens to broadly explore gangliosides that react to serum. All the acoustic neurinoma samples used in the present study showed a similar ganglioside profile on TLC (thin-layer chromatography). For the microscale ganglioside analysis, a newly developed TLC blotting/secondary ion mass spectrometry (SIMS) system together with TLC immunostaining method was employed. Most of the ganglioside bands could be analyzed, and they were identified as GM3, GM2, SPG, GM1a, GD3, S i (sialyl-i ganglioside) and GD1a. GD1a was the predominant ganglioside and many neolactoseries gangliosides were recognized by immunological analysis. Next, the immune reactivity of serum samples, from patients with Meniere's disease, with the acoustic neurinoma gangliosides was studied by TLC immunostaining. The result showed that five of 11 patients with Meniere's disease and one of eight normal subjects reacted with a specific band, which was identified as S-i by the TLC blotting/SIMS system. The findings of the present study indicate that S-i ganglioside is an autoantigen and possibly involved in the pathogenesis of Meniere's disease. PMID- 10838183 TI - Inflammatory response following acute magnesium deficiency in the rat. AB - The importance of inflammatory processes in the pathology of Mg deficiency has been recently reconsidered but the sequence of events leading to the inflammatory response remains unclear. Thus, the purpose of the present study was to characterize more precisely the acute phase response following Mg deficiency in the rat. Weaning male Wistar rats were pair-fed either a Mg-deficient or a control diet for either 4 or 8 days. The characteristic allergy-like crisis of Mg deficient rats was accompanied by a blood leukocyte response and changes in leukocytes subpopulations. A significant increase in interleukin-6 (IL-6) plasma level was observed in Mg-deficient rats compared to rats fed a control diet. The inflammatory process was accompanied by an increase in plasma levels of acute phase proteins. The concentrations of alpha2-macroglobulin and alpha1-acid glycoprotein in the plasma of Mg-deficient rats were higher than in control rats. This was accompanied in the liver by an increase in the level of mRNA coding for these proteins. Moreover, Mg-deficient rats showed a significant increase in plasma fibrinogen and a significant decrease in albumin concentrations. Macrophages found in greater number in the peritoneal cavity of Mg-deficient rats were activated endogenously and appeared to be primed for superoxide production following phorbol myristate acetate stimulation. A high plasma level of IL-6 could be detected as early as day 4 for the Mg-deficient diet. Substance P does not appear to be the initiator of inflammation since IL-6 increase was observed without plasma elevation of this neuropeptide. The fact that the inflammatory response was an early consequence of Mg deficiency suggests that reduced extracellular Mg might be responsible for the activated state of immune cells. PMID- 10838184 TI - Phosphatidylserine-dependent adhesion of T cells to endothelial cells. AB - Phosphatidylserine (PS) was exposed at the surface of human umbilical vein endothelial cells (HUVECs) and cultured cell lines by agonists that increase cytosolic Ca(2+), and factors governing the adhesion of T cells to the treated cells were investigated. Thrombin, ionophore A23187 and the Ca(2+)-ATPase inhibitor 2, 5-di-tert-butyl-1,4-benzohydroquinone each induced a PS-dependent adhesion of Jurkat T cells. A23187, which was the most effective agonist in releasing PS-bearing microvesicles, was the least effective in inducing the PS dependent adhesion of Jurkat cells. Treatment of ECV304 and EA.hy926 cells with EGTA, followed by a return to normal medium, resulted in an influx of Ca(2+) and an increase in adhering Jurkat cells. Oxidised low-density lipoprotein induced a procoagulant response in cultured ECV304 cells and increased the number of adhering Jurkat cells, but adhesion was not inhibited by pretreating ECV304 cells with annexin V. PS was not significantly exposed on untreated Jurkat cells, as determined by flow cytometry with annexin V-FITC. However, after adhesion to thrombin-treated ECV304 cells for 10 min followed by detachment in 1 mM EDTA, there was a marked exposure of PS on the Jurkat cells. Binding of annexin V-FITC to the detached cells was inhibited by pretreating them with unlabelled annexin V. Contact with thrombin-treated ECV304 cells thus induced the exposure of PS on Jurkat cells and, as Jurkat cells were unable to adhere to thrombin-treated ECV304 cells in the presence of EGTA, the adhesion of the two cell types may involve a Ca(2+) bridge between PS on both cell surfaces. The number of T cells from normal, human peripheral blood that adhered to ECV304 cells was not increased by treating the latter with thrombin. However, findings made with several T cell lines were generally, but not completely, consistent with the possibility that adhesion to surface PS on endothelial cells may be a feature of T cells that express both CD4(+) and CD8(+) antigens. Possible implications for PS-dependent adhesion of T cells to endothelial cells in metastasis, and early in atherogenesis, are discussed. PMID- 10838185 TI - Apparent cooperativity in multivalent verotoxin-globotriaosyl ceramide binding: kinetic and saturation binding studies with [(125)I]verotoxin. AB - Verotoxin (VT) binding to the trisaccharide portion of globotriaosyl ceramide (Gb(3)) is believed to be a crucial step in the development of hemolytic uremic syndrome (HUS) commonly known as 'Hamburger disease'. This interaction is the initial step in the binding process and defines the specificity of verotoxin binding to cellular membranes. Although molecular modeling, co-crystallization and co-NMR studies with VT and the trisaccharide moiety of Gb(3) have indicated potential multiple sites for Gb(3) binding, little is known about their direct effects on kinetic and equilibrium binding. Here we describe how the binding of radiolabeled VT ([(125)I]VT1) to Gb(3) in a microtiter well format, is driven by two different association rate constants (k(+1a)=0.0075 and k(+1b)=0.275 min(-1) nM(-1)) with the high affinity site representing 15% of the total specific binding sites. Binding was reversible at room temperature, reached equilibrium after 2-3 h, and non-specific binding was less than 5%. Equilibrium binding studies defined by [(125)I]VT1 saturation binding to 15, 30, 60 and 120 ng Gb(3)/well, showed the presence of a single site with dissociation constants (K(d)s) ranging between 0.5 and 3 nM. However, the maximum density of specific [(125)I]VT1 binding sites (B(max)) did not directly correlate with the Gb(3) concentration per well: the most[(125)I]VT1 binding was observed for 60 ng Gb(3) (B(max)=1.28 nM; compared to 0. 23 nM for 30 ng Gb(3) and 0.65 nM for 120 ng Gb(3)). Furthermore, while Hill coefficients (n(H)) for 15, 30 and 120 ng Gb(3) were close to unity indicating single interactions, for the saturation isotherm for 60 ng Gb(3)/well n(H) was 1.4. Subsequent Scatchard analysis yielded a concave downward curve for [(125)I]VT1 binding to 60 ng Gb(3)/well, suggesting positive co-operativity. We present, for the first time, conclusive binding data confirming the presence of at least two discrete Gb(3) binding sites: these multivalent interactions between verotoxin VT-1 and Gb(3) were described by association reactions driven by two distinct rate constants, as well as by the positive co-operativity governing binding at a restricted receptor concentration. These results imply that the concentration of Gb(3) on the surface of target cells can have a complex, non-linear effect on verotoxin binding and thereby, on sensitivity to cytotoxicity. PMID- 10838186 TI - Identification of a novel aspartic-like protease differentially expressed in human breast cancer cell lines. AB - Four different human breast cancer cell lines were examined to search for genes associated with tumor growth and metastasis. Each of these cell lines, MDA-MB 453, MCF-7, MDA-MB-231 and MDA-MB-435, displays different phenotypic characteristics ranging from poorly to highly tumorigenic and metastatic. The differences in gene expression profiles of these cell lines generated by differential display technique should allow one to identify candidates as putative oncogenes or tumor/metastasis suppressor genes. A novel cDNA expressed in the highly tumorigenic and metastatic cell line, MDA-MB-435, was identified and isolated by this approach. The function for this gene, designated ALP56 (aspartic-like protease 56 kDa), in tumor progression is suggested by the homology of the encoded protein to aspartic proteases, such as cathepsin D. The amino acid residues in two catalytic domains of this family are highly conserved in those domains of ALP56. Northern hybridization indicated that the expression of ALP56 is associated with growth and metastasis of MDA-MB-435 tumors in immunodeficient mice. In situ hybridization of biopsies from breast cancer and colon cancer patients indicated that ALP56 is upregulated in human primary tumors and liver metastasis. These results suggest that this novel gene correlates with human tumor progression. PMID- 10838187 TI - Iron overload and kidney lysosomes. AB - Iron overload has been associated with damage of the liver and other organs of patients with primary or secondary increased iron load. In order to study the effect of iron overload on the pathophysiology of kidney lysosomes, experimentally induced iron overload models were employed. Iron overload was achieved through intraperitoneal injections of Fe-dextran (Imferon) in male rats, at different final iron concentrations (825 and 1650 mg/kg, single and double dose groups respectively). Controls were injected with dextran following a similar protocol. The animals were killed at different time points after the last injection. Subcellular fractionation studies of kidney homogenates were carried out by differential centrifugation and density gradient centrifugation. The kidney iron load was increased with both doses. Iron appeared to accumulate mainly in the lysosomes, bringing about distinct changes in the behaviour of the organelles as judged by subcellular fractionation studies. Lysosomes became more fragile and showed increased density. The extent of the above changes seemed to correlate with the extent and duration of iron accumulation and could be reversed when the iron load was reduced. PMID- 10838188 TI - The pyrrolopyrimidine U101033E is a potent free radical scavenger and prevents Fe(II)-induced lipid peroxidation in synaptosomal membranes. AB - The pyrrolopyrimidine U101033E is a therapeutic compound potentially useful in stroke, head injury and other oxidative stress conditions. Electron paramagnetic resonance (EPR) techniques of spin labeling and spin trapping in conjunction with measures of lipid and protein oxidation have been used to investigate the proposed antioxidant capacity of U101033E. We report potent antioxidant activity of this agent in aqueous cell-free solution as measured by spin trapping. U101033E significantly (P<0.005) reduces the formation of the EPR active spin trap N-t-butyl-alpha-phenylnitrone (PBN)-radical adduct by 17.1% at a concentration of 1 microM, four orders of magnitude less than the concentration of PBN. As measured by the decrease in signal intensity of lipid-resident nitroxide stearate spin probes, an EPR assay for lipid peroxidation, this pyrrolopyrimidine compound efficiently protected against hydroxyl radical-induced lipid peroxidation in cortical synaptosomal membranes deep within the membrane bilayer, but not closer to the membrane surface. In addition, U101033E partially prevents synaptosomal protein oxidation in the presence of Fe(II); however, U101033E demonstrates some protein oxidative effects itself. These results are supportive of the proposed role of U101033E as a lipid-specific antioxidant, especially for protection against lipid peroxidation that occurs deep within the membrane bilayer, but raise some potential concerns about the oxidative nature of this agent toward proteins. PMID- 10838189 TI - Colorectal cancer-associated nuclear antigen. AB - By using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting assays in the presence of polyclonal antiserum raised against electrophoretically specific polypeptides of colorectal cancer nuclear polypeptides with M(r) of 35-40 kDa, we have identified p36 protein whose expression accompanies tumorigenesis of large intestine. Immunological analysis of 35 nuclear protein preparations has indicated expression of p36 antigen in nine of 11 right-sided (81.8%) and 21 of 24 (87.5%) left-sided colorectal tumor cases, but not in any control tissue samples. In this study, we have identified p36 antigen in two colon tumor cell lines, i.e., SW620 and HT29 as well. Fractionation experiments based on selective extraction of nuclei isolated from cancerous specimens, which enables their separation into chromatin, nuclear matrix and its subfraction, i.e., internal and peripheral matrix have revealed the concentration of this particular antigen in the internal matrix. PMID- 10838190 TI - Stimulus- and cell-type-specific regulation of CCAAT-enhancer binding protein isoforms in glomerular mesangial cells by lipopolysaccharide and cytokines. AB - Binding sites for the CCAAT-enhancer binding protein (C/EBP) family are present in the promoter regions of several genes that are known to be expressed by mesangial cells (MC) during the pathogenesis of glomerular inflammatory diseases. The precise regulation of the C/EBP family by agents that are known to activate MC is, however, poorly understood. We report here the action of interleukin-1 (IL)-1 and, for the first time, lipopolysaccharide (LPS), platelet-derived growth factor (PDGF), IL-6, interferon-gamma (IFN-gamma) and tumour necrosis factor alpha (TNF-alpha) on the C/EBP expression profile and functional DNA binding activity in primary rat MC. Both cell-type- and stimulus-specific regulation of C/EBP mRNA expression and DNA binding activity were identified, with C/EBPalpha being induced by LPS, C/EBPbeta by LPS, IL-1, TNF-alpha and C/EBPdelta by LPS, IL 1, IFN-gamma, TNF-alpha and PDGF. Such differential regulation, particularly that of C/EBPbeta, may be responsible for the mediator-specific differences in the expression of C/EBP-regulated genes in MC. Additionally, the involvement of potential post-transcriptional mechanisms in the regulation of C/EBPdelta were identified. These studies provide novel insights into the stimulus-specific regulation of gene expression during renal diseases. PMID- 10838191 TI - Expression patterns of chondrocyte genes cloned by differential display in tibial dyschondroplasia. AB - Tibial dyschondroplasia (TD) appears to involve a failure of the growth plate chondrocytes within growing long bones to differentiate fully to the hypertrophic stage, resulting in a mass of prehypertrophic chondrocytes which form the avascular TD lesion. Many biochemical and molecular markers of chondrocyte hypertrophy are absent from the lesion, or show reduced expression, but the cause of the disorder remains to be identified. As differentiation to the hypertrophic state is impaired in TD, we hypothesised that chondrocyte genes that are differentially expressed in the growth plate should show altered expression in TD. Using differential display, four genes, B-cadherin, EF2, HT7 and Ex-FABP were cloned from chondrocytes stimulated to differentiate to the hypertrophic stage in vitro, and their differential expression confirmed in vivo. Using semi quantitative RT-PCR, the expression patterns of these genes were compared in chondrocytes from normal and TD growth plates. Surprisingly, none of these genes showed the pattern of expression that might be expected in TD lesion chondrocytes, and two of them, B-cadherin and Ex-FABP, were upregulated in the lesion. This indicates that the TD phenotype does not merely reflect the absence of hypertrophic marker genes, but may be influenced by more complex developmental mechanisms/defects than previously thought. PMID- 10838192 TI - Detection of pathological molecular alterations in scrapie-infected hamster brain by Fourier transform infrared (FT-IR) spectroscopy. AB - In this report a new approach for the identification of pathological changes in scrapie-infected Syrian hamster brains using Fourier transform infrared microspectroscopy is discussed. Using computer-based pattern recognition techniques and imaging, infrared maps with high structural contrast were obtained. This strategy permitted comparison of spectroscopic data from identical anatomical structures in scrapie-infected and control brains. Consistent alterations in membrane state-of-order, protein composition, carbohydrate and nucleic acid constituents were detected in scrapie-infected tissues. Cluster analysis performed on spectra of homogenized medulla oblongata and pons samples also reliably separated uninfected from infected specimens. This method provides a useful tool not only for the exploration of the disease process but also for the development of rapid diagnostic and screening techniques of transmissible spongiform encephalopathies. PMID- 10838193 TI - Metabolism of carnitine in phenylacetic acid-treated rats and in patients with phenylketonuria. AB - The effect of metabolites accumulating in phenylketonuria (PKU) was investigated on carnitine metabolism in rats and in patients with PKU. Of phenylacetic acid (PEAA), phenylpyruvic acid and homogentisic acid the PEAA was found to be the most effective in inhibiting carnitine biosynthesis in rats. Following 60 min, a single intraperitoneal dose of PEAA the relative conversion rate, i. e. the hydroxylation, of tracer [Me-(3)H]butyrobetaine to [Me-(3)H]carnitine decreased from 62.2+/-6.00% to 39.4+/-5.11% (means+/-S.E.M., P<0.01) in the liver, in the only organ doing this conversion in rats. The conversion of loading amount of unlabeled butyrobetaine to carnitine was also markedly reduced. The impaired hydroxylation of butyrobetaine was reflected by a reduced free and total carnitine levels in the liver and a reduced total carnitine concentration in the plasma. PEAA decreased the hepatic level of glutamic acid and alpha-ketoglutaric acid (alpha-KG), suggesting a mechanism for the reduced flux through the butyrobetaine hydroxylase enzyme, because alpha-KG is an obligatory co-enzyme. In the plasma and urine of PKU patients on unrestricted diet, markedly decreased total carnitine levels were detected. In the liver of PEAA-treated rats and urine of PKU patients, a novel carnitine derivative, phenacetyl-carnitine was verified by HPLC and gas chromatography-mass spectrometry. PMID- 10838194 TI - The dissociation between upregulated endothelins and hemodynamic responses during polymicrobial sepsis. AB - Polymicrobial sepsis is characterized by an early, hyperdynamic phase followed by a late, hypodynamic phase. Although studies have suggested that endothelins (ETs) contribute to the development of shock after a bolus injection of endotoxin, little is known about the role of ETs in the transition from the hyperdynamic phase to the hypodynamic phase of sepsis. To study this, male adult rats were subjected to sepsis by cecal ligation and puncture (CLP) followed by fluid resuscitation. Plasma levels of ET-1 and ET-2 were measured by radioimmunoassay at 2, 5, 10 h (i.e. the early stage of sepsis), and 20 h (late stage) following CLP or sham operation. Tissue levels of ET-1 and ET-2 were determined in the heart, lungs, small intestine, and spleen at 5 h after CLP or sham operation. In addition, preproendothelin-1 (precursor of ET-1) gene expression was analyzed by reverse transcription-polymerase chain reaction (RT-PCR) at 5 h in the heart, lungs, small intestine, spleen, and liver. The results indicate that plasma levels of ET-1 and ET-2 were not different from values of sham groups at 2 and 20 h, but were significantly higher than the sham values at 5 and 10 h after CLP. While there were no significant increases in tissue levels of ET-1 and ET-2 at 5 h post-CLP, RT-PCR analysis indicates a significant upregulation of preproendothelin-1 gene expression in the heart, spleen, and liver (but not in the lungs or small intestine) at 5 h after the onset of sepsis. These results indicate that the heart, spleen, and liver appear to be important ET-producing organs during the early stage of sepsis. The lack of significant increases in tissue ET levels could be due to the possibility that the newly converted peptide is quickly transferred to the bloodstream. Since the hyperdynamic phase of sepsis occurs at 2-10 h and the hypodynamic phase occurs at 20 h after CLP, the increased plasma levels of ET at 5 and 10 h suggest that mediators other than ETs (such as adrenomedullin) are responsible for producing the biphasic hemodynamic responses during the progression of polymicrobial sepsis. PMID- 10838195 TI - Characterization of an 84 kDa protein inducing an immediate hypersensitivity reaction in rabbits sensitized to Haemaphysalis longicornis ticks. AB - An immunogenic 84 kDa protein was isolated and purified from whole tick extracts of Haemaphysalis longicornis larvae by a combination of ion exchange, reverse phase and hydrophobic interaction chromatographies. The protein, when injected intradermally into rabbits exposed to repeated tick feeding, induces an immediate cutaneous hypersensitivity reaction. It has been purified to homogeneity as shown by sodium dodecyl sulphate polyacrylamide gel electrophoresis and silver staining. Amino acid sequences for two peptides derived from proteolytic cleavage of p84 were scanned against known proteins on the SWISS-PROT database. A 7 residue motif, ISGWGNT present in one of the two peptides appeared conserved in both vertebrate and invertebrate trypsin-like serine proteinases, while another 7 amino acid motif, HVPAGQI present in the second peptide showed homology to an Escherichia coli ATP-binding protein. We have discussed our findings in relation to isolation and characterization of target antigens for tick vaccine candidates. PMID- 10838196 TI - Characterization of two alpha-galactosidase mutants (Q279E and R301Q) found in an atypical variant of Fabry disease. AB - The mutant products Q279E ((279)Gln to Glu) and R301Q ((301)Arg to Gln) of the X chromosomal inherited alpha-galactosidase (EC 3.2.1. 22) gene, found in unrelated male patients with variant Fabry disease (late-onset cardiac form) were characterized. In contrast to patients with classic Fabry disease, who have no detectable alpha-galactosidase activity, atypical variants have residual enzyme activity. First, the properties of insect cell-derived recombinant enzymes were studied. The K(m) and V(max) values of Q279E, R301Q, and wild-type alpha galactosidase toward an artificial substrate, 4-methylumbelliferyl-alpha-D galactopyranoside, were almost the same. In order to mimic intralysosomal conditions, the degradation of the natural substrate, globotriaosylceramide, by the alpha-galactosidases was analyzed in a detergent-free-liposomal system, in the presence of sphingolipid activator protein B (SAP-B, saposin B). Kinetic analysis revealed that there was no difference in the degradative activity between the mutants and wild-type alpha-galactosidase activity toward the natural substrate. Then, immunotitration studies were carried out to determine the amounts of the mutant gene products naturally occurring in cells. Cultured lymphoblasts, L-57 (Q279E) and L-148 (R301Q), from patients with variant Fabry disease, and L-20 (wild-type) from a normal subject were used. The 50% precipitation doses were 7% (L-57) and 10% (L-148) of that for normal lymphoblast L-20, respectively. The residual alpha-galactosidase activity was 3 and 5% of the normal level in L-57 and L-148, respectively. The quantities of immuno cross reacting materials roughly correlated with the residual alpha-galactosidase activities in lymphoblast cells from the patients. Compared to normal control cells, fibroblast cells from a patient with variant Fabry disease, Q279E mutation, secreted only small amounts of alpha-galactosidase activity even in the presence of 10 mM NH(4)Cl. It is concluded that Q279E and R301Q substitutions do not significantly affect the enzymatic activity, but the mutant protein levels are decreased presumably in the ER of the cells. PMID- 10838197 TI - Increased cytogenetic damage in outdoor painters. AB - Painters are exposed to an extensive variety of hazardous substances such as organic solvents, lead-containing pigments and residual plastic monomers. In this particular case, workers used commercially available exterior paints and occasionally gasoline or thinner as solvents. The application or removal of paints was performed without protection (masks or gloves). To determine occupational exposure risk, a monitoring study was designed. Group selection was made after a questionnaire administration, which included questions about lifestyle and medical history to exclude exposure to other potential sources of genotoxics. Smoking and drinking habits were also considered. Blood and buccal cell samples were obtained from 25 public building male painters and from a similar number of age- and gender-matched controls. Lead levels were measured in paint samples and in individuals' blood. Organic solvents and/or its metabolites were also determined in blood. Chromosomal aberrations (CA) and sister chromatid exchanges (SCE) were determined in peripheral blood lymphocyte cultures. Also, the frequency of micronuclei (MN) in buccal cells was investigated. Painters had higher lead levels in blood (p<0.05); CA and SCE in lymphocytes and MN in epithelial cells were also elevated (p<0.05). Cytogenetic damage was significantly associated with occupational exposure time but not with the levels of lead found in blood. PMID- 10838198 TI - Chlorophyllin [CHLN] and the mutagenicity of monofunctional alkylating agents in Drosophila: the action of CHLN need not include an influence on metabolic activation. AB - The effect of chlorophyllin (CHLN) on the mutagenicity of four monofunctional alkylating agents (MFAAs) was evaluated in the wing spot test in Drosophila. Three of the compounds are direct-acting (ethylnitrosamine (ENU), methylnitrosourea (MNU), and methylmethanesulfonate (MMS)) and one indirect acting (diethylnitrosamine, DEN). Results indicate that the mutagenicity of all four compounds is strongly inhibited by CHLN. The findings are not in agreement with the conclusion of Romert et al. (1992) that CHLN has no effect on the mutagenicity of direct acting MFFAs inferred from their work with MNU and ethylmethanesulfonate (EMS) in the V79 and Salmonella in vitro test systems. The results suggest the possibility that the action of CHLN need not include an inhibiting effect on metabolic activation. PMID- 10838199 TI - Mutagenic evaluation of deltamethrin using rodent dominant lethal assay. AB - The dominant lethal test was used to analyse the mutagenic potential of deltamethrin, a synthetic pyrethroid insecticide, in Swiss albino mice. In the treated series, the animals were exposed orally to three different doses (0.36, 0.72 and 1.08 mg/kg body weight) of deltamethrin dissolved in corn oil. Following the treatment, each male of control, as well as of the treated series, was mated with untreated females, every week for a period of 6 weeks. All mated females were sacrificed on the 13th day of separation and their ovaries and uterus were examined. The results revealed that deltamethrin treatment did not impair the mating capacity and fertility of Swiss albino mice. Mutagenic index, pre- and post-implantation losses were assessed. No significant pre-implantation losses were observed either weekly or averagely. Post-implantation losses were observed at medium and high doses of deltamethrin. A slight increase in dominant lethal mutation rate was observed by increasing doses of deltamethrin in early weeks but decreased in later weeks, so an apparent dose response was not observed. PMID- 10838200 TI - Polymyxin permeabilization as a tool to investigate cytotoxicity of therapeutic aromatic alkylators in DNA repair-deficient Escherichia coli strains. AB - Chlorambucil (CLB; N,N-bis(2-chloroethyl)-p-aminophenylbutyric acid) and its biologically active beta-oxidation product phenylacetic acid mustard (PAM; N,N bis(2-chloroethyl)-p-aminophenylacetic acid) are bifunctional aromatic alkylators. CLB is in wide clinical use as an anticancer drug and also as an immunosuppressant. The chemical structures indicate that CLB and PAM are mutagenic, teratogenic and carcinogenic, but the mode of action has remained obscure. We have investigated the biological effects of CLB and PAM with DNA repair-deficient Escherichia coli strains. In contrast to MNNG (N-methyl-N'-nitro N-nitrosoguanine), CLB and PAM were not toxic to E. coli, but permeabilization of the outer membrane of the cells through use of polymyxin B nonapeptide (PMBN) rendered them susceptible to these compounds. The importance of DNA repair, shown by reversal of damage and attenuation of the toxicity of CLB and PAM, was indicated by the susceptibility of cells lacking O(6)-methylguanine-DNA methyltransferase I and II (ada ogt). Similarly, the protective role of base excision repair (BER) was substantiated by demonstration of an even more increased susceptibility to CLB and PAM of cells lacking 3-methyladenine-DNA glycosylase I and II (alkA1 tag-1). Cells deficient in mismatch repair (mutS) appeared to be slightly more sensitive than normal cells to CLB and PAM, although no such sensitivity to MNNG was observed. This implicates the role of mismatches in CLB- and PAM-related cytotoxicity. It is generally believed that bifunctional alkylating agents, like CLB and PAM, exert their cytotoxic action via DNA cross linking. Our results with O(6)-methyltransferase- and 3-methyladenine-DNA glycosylase-deficient cells indicate that removal of the adducts prior to the formation of cross-links is an important mechanism maintaining cell viability. We conclude that PMBN permeabilization provides a valuable tool to investigate genetically engineered E. coli cells, whose outer membrane is not naturally permeable to mutagens or other interesting compounds. PMID- 10838201 TI - Studies on a novel DNA polymerase inhibitor group, synthetic sulfoquinovosylacylglycerols: inhibitory action on cell proliferation. AB - Some chemically synthesized sulfoquinovosylmonoacylglycerols (SQMG) sulfoquinovosyldiacylglycerols (SQDG) have been reported to selectively and strongly inhibit the activities of mammalian DNA polymerases alpha and beta in vitro. In this study, using human cancer cell lines, we investigated the effects of SQMG-SQDG on the DNA polymerase in the cells. In the presence of n-decane, the IC(50) values on cell growth were approximately 1-5 microM for SQMG and about 0.3 1 microM for SQDG. The values were almost the same as the in vitro enzyme inhibitory levels. The cell lines were arrested in early S-phase by SQMG-SQDG at the concentrations of 0.1-4.7 microM in a manner dependent on incubation time, suggesting that SQMG-SQDG blocked the primary step of DNA replication by inhibiting DNA polymerase, possibly alpha-type. We also demonstrated the localization of SQMG in the cell using the fluorescent SQMG analog, SQMGalpha NBDD, which was synthesized in our laboratory. SQMGalpha-NBDD was localized in the nucleus and on the nuclear surface, but the binding site seemed not to be the DNA/chromatin, suggesting that the SQMG-SQDG might interact with molecules located close to the DNA/chromatin and on the nuclear surface. These results suggested a correlation between the in vitro biochemical action of the SQMG-SQDGs and their intracellular mode of action. PMID- 10838202 TI - Individual variation in somatic mutations of the glycophorin-A gene in neonates in relation to pre-natal factors. AB - The frequency of somatic mutations of the glycophorin-A (GPA) gene was measured in red blood cells from a series of newborn babies and related to various epidemiological and lifestyle factors in order to identify those factors that might influence the mutation rate before birth. Although there was substantial variation in the mutation frequencies between individual babies, no specific associations were found with any of the factors explored including smoking, age and social class of the parents, and gender and birth weight of the babies. It is concluded that these factors do not have a substantial effect on the mutational endpoint measured, although this does not necessarily mean that they have no effect on health risk to the offspring. The observed variation in GPA gene mutation frequency must, therefore, be due either to exposure to less obvious external influences or to intrinsic factors. PMID- 10838203 TI - Genotoxic potential of marine sediments from the North Sea. AB - The alkaline comet assay is a method for detecting DNA strand breaks and alkali labile sites in individual cells. An in vitro system was used to investigate the genotoxic potential of complex mixtures such as organic extracts of marine sediments. DNA damage was induced in leukocytes isolated from carp (Cyprius carpio) by exposure to organic sediment extracts from the North Sea or hydrogen peroxide as positive control, respectively. The minimum concentration for significant effects ranged from 1 to 40 mg sediment dry weight per milliliter assay volume. The sensitivity of the method was enhanced by using the DNA repair inhibitor, 1-beta-D-arabinofuranosylcytosine (ara C). From the results, it can be suggested that total organic carbon (TOC) as well as the different compositions of contaminants present in the sediment extracts may contribute to the genotoxic effects observed. The comet assay can be applied successfully as an in vitro bioassay for investigations on genotoxicity of marine sediment extracts. PMID- 10838204 TI - Chromosome aberration analysis in Concorde pilots. AB - Chromosomal aberrations, micronuclei, and sister chromatid exchanges have been analysed in human peripheral lymphocytes of 18 Concorde pilots and 10 controls. There was an eightfold significant increase of dicentric chromosomes in the Concorde group. The yield of micronuclei was also significantly elevated. Sister chromatid exchanges in the Concorde group did not differ from the control. Comparing the results to flight personnel from subsonic routes, the dicentric yield was higher in personnel from supersonic crews but the difference was not statistically significant. The overdispersion of dicentric chromosomes showed the influence of high LET cosmic radiation. The estimated mean dose per year ranged from 11 to 37 mSv depending on the radiation weighting factor for neutrons. It is recommended that actual and future high-speed transport should consider not only physical measurements, but also biological data like the frequencies of chromosomal aberrations because the latter reflect sensitively the high biological effectiveness of cosmic radiation. PMID- 10838205 TI - Induction of micronuclei and other nuclear abnormalities in European minnow Phoxinus phoxinus and mollie Poecilia latipinna: an assessment of the fish micronucleus test. AB - In this work, we have measured both micronuclei and other nuclear abnormalities in renal erythrocytes from European minnow Phoxinus phoxinus and mollie Poecilia latipinna, with the aim to contribute to the standardisation of the micronucleus test for fish species. Intraperitoneal injections of colchicine (10 mg/kg), cyclophosphamide (40 mg/kg), or mitomycin C (20 mg/kg) for 24 h induced diverse nuclear abnormalities in minnow erythrocytes, therefore nuclear abnormalities should be added to micronuclei as genotoxicity indicators in fish micronucleus tests. The adequacy of administration protocols based on intraperitoneal injections has been evaluated by injecting saline solution to both species: single or double injections have not induced neither micronuclei nor other nuclear abnormalities in any case. Finally, the differential sensitivity of both species to toxic heavy metals was evaluated by exposing individuals of both species to different doses (0.17, 1.7, 2x1.7, and 3.4 mg/kg) of cadmium and mercury for 24 h; we concluded that the mollie is sensitive to both metals whereas the minnow is not sensitive to mercury. PMID- 10838206 TI - The origins of the back-mutation assay method: a personal recollection. AB - The back-mutation assay method for determining the mutagenicity of various treatments was first developed a little over 50 years ago and has been in continuous use ever since. Shortly after the method was first used it became evident that certain factors of cell density, composition of media, etc., had to be carefully controlled to preserve an acceptable reliability of the method. A factor of particular importance was the suppression of growth of back-mutant prototrophic cells by the large number of auxotrophic cells present, a phenomenon which later became known as the "Grigg Effect." This review describes the origins of the back-mutation method and of the confounding competitive suppression phenomenon, the cause of competitive suppression, methods of diagnosing whether it is likely to bias the interpretation of a particular back-mutation experiment, and an experimental design which removes it entirely as a possible source of error. A number of other phenomena, such as phenotypic lag and coincident mutation associated with back-mutation, are also discussed as possible sources of error. PMID- 10838207 TI - Single cell gel electrophoresis assay: a new technique for human biomonitoring studies. AB - Human biomonitoring using the single cell gel electrophoresis (SCGE) or comet assay is a novel approach for the assessment of genetic damage in exposed populations. This assay enables the detection of various forms of DNA damage in individual cells with ease and speed and is, therefore, well suited to the analysis of a large group in a population. Here, application of SCGE assay in the identification of dietary protective factors, in clinical studies and in monitoring the risk of DNA damage resulting from occupational, environmental or lifestyle exposures is reviewed. Also, the comparative sensitivity of SCGE assay and conventional cytogenetic tests to detect genetic damage is discussed. Finally, strengths and shortcomings of the SCGE assay are addressed. PMID- 10838208 TI - Effects of chemical contaminants on genetic diversity in natural populations: implications for biomonitoring and ecotoxicology. AB - The conservation of genetic diversity has emerged as one of the central issues in conservation biology. Although researchers in the areas of evolutionary biology, population management, and conservation biology routinely investigate genetic variability in natural populations, only a handful of studies have addressed the effects of chemical contamination on population genetics. Chemical contamination can cause population reduction by the effects of somatic and heritable mutations, as well as non-genetic modes of toxicity. Stochastic processes in small populations, increased mutation load, and the phenomenon of mutational meltdown are compounding factors that cause reduced fitness and accelerate the process of population extirpation. Although the original damage caused by chemical contaminants is at the molecular level, there are emergent effects at the level of populations, such as the loss of genetic diversity, that are not predictable based solely on knowledge of the mechanism of toxicity of the chemical contaminants. Therefore, the study of evolutionary toxicology, which encompasses the population-genetic effects of environmental contaminants, should be an important focus of ecotoxicology. This paper reviews the issues surrounding the genetic effects of pollution, summarizes the technical approaches that can be used to address these issues, and provides examples of studies that have addressed some of them. PMID- 10838209 TI - Evaluation of fecal mutagenicity and colorectal cancer risk. AB - Colorectal cancer is one of the most common internal malignancies in Western society. The cause of this disease appears to be multifactorial and involves genetic as well as environmental aspects. The human colon is continuously exposed to a complex mixture of compounds, which is either of direct dietary origin or the result of digestive, microbial and excretory processes. In order to establish the mutagenic burden of the colorectal mucosa, analysis of specific compounds in feces is usually preferred. Alternatively, the mutagenic potency of fecal extracts has been determined, but the interpretation of these more integrative measurements is hampered by methodological shortcomings. In this review, we focus on exposure of the large bowel to five different classes of fecal mutagens that have previously been related to colorectal cancer risk. These include heterocyclic aromatic amines (HCA) and polycyclic aromatic hydrocarbons (PAH), two exogenous factors that are predominantly ingested as pyrolysis products present in food and (partially) excreted in the feces. Additionally, we discuss N nitroso-compounds, fecapentaenes and bile acids, all fecal constituents (mainly) of endogenous origin. The mutagenic and carcinogenic potency of the above mentioned compounds as well as their presence in feces, proposed mode of action and potential role in the initiation and promotion of human colorectal cancer are discussed. The combined results from in vitro and in vivo research unequivocally demonstrate that these classes of compounds comprise potent mutagens that induce many different forms of genetic damage and that particularly bile acids and fecapentaenes may also affect the carcinogenic process by epigenetic mechanisms. Large inter-individual differences in levels of exposures have been reported, including those in a range where considerable genetic damage can be expected based on evidence from animal studies. Particularly, however, exposure profiles of PAH and N-nitroso compounds (NOC) have to be more accurately established to come to a risk evaluation. Moreover, lack of human studies and inconsistency between epidemiological data make it impossible to describe colorectal cancer risk as a result of specific exposures in quantitative terms, or even to indicate the relative importance of the mutagens discussed. Particularly, the polymorphisms of genes involved in the metabolism of heterocyclic amines are important determinants of carcinogenic risk. However, the present knowledge of gene-environment interactions with regard to colorectal cancer risk is rather limited. We expect that the introduction of DNA chip technology in colorectal cancer epidemiology will offer new opportunities to identify combinations of exposures and genetic polymorphisms that relate to increased cancer risk. This knowledge will enable us to improve epidemiological study design and statistical power in future research. PMID- 10838210 TI - Immunogenetic influences on tick resistance in African cattle with particular reference to trypanotolerant N'Dama (Bos taurus) and trypanosusceptible Gobra zebu (Bos indicus) cattle. AB - In sub-Saharan Africa, tick infestation and tick-borne infections together with tsetse-transmitted trypanosomosis arguably constitute the main parasitological disease complex constraining livestock production. Resistance to tick attack and tick-borne micro-organisms (TBMs) varies among different breeds of cattle. The magnitude of losses due to these parasites is related to an extent to the degree of breed resistance. Generally, zebu (Bos indicus) cattle possess a higher resistance to ticks and TBMs than European (Bos taurus) cattle. The host's immune system would appear to be the single most important factor that regulates this resistance. This paper reports on the main effector immune mechanisms governing resistance against ticks and TBMs. The cellular immune response appears more effective and stable than humoral immunity in modulating resistance to ticks and TBMs. Similarities between the immune mechanisms employed by trypanotolerant N'Dama (B. taurus) cattle, when infected with trypanosomes, and those elicited by tick bites and TBMs seem to exist, particularly at the skin level in the early phases of parasitic invasion. Moreover, there is evidence that in the N'Dama breed, resistance against ticks per se also has a genetic basis. Therefore, the N'Dama appears to be a unique breed in that it exhibits resistance to several parasitic diseases and/or infections, including helminths, when compared to other cattle breeds in West Africa. It is concluded that the multi-parasite resistant traits of the N'Dama breed should be exploited in those areas where trypanosomosis, ticks and tick-borne diseases constrain animal production. This should be of benefit for low-input farming systems where the use of chemicals for prophylaxis and therapy is limited by their relatively high cost. Additionally, the potential contribution of multiple disease resistant N'Dama cattle should be considered in crossbreeding programmes with exotic dairy breeds for increasing milk production in West Africa. PMID- 10838211 TI - Measuring exposure to Schistosoma japonicum in China. III. Activity diaries, snail and human infection, transmission ecology and options for control. AB - We used activity diaries and snail detection to relate water contact and Schistosoma japonicum infection among a cohort of 178 residents on two islands in the Dongting Lake, China. Water exposure to each of 12 mapped water zones around the islands was calculated (m(2) min/day) for each subject. Infected Oncomelania hupensis hupensis snails in this area are focal and were found in only five of the 12 zones, with the highest rate being 5.7%. Thirty-one subjects (17%) were re infected with a mean intensity of 63.2 epg. Mean water contact was 7.9 m(2) min/day; 98% of water exposure was due to economic activity and only 2% due to swimming or bathing, washing and other necessities of daily life. Males had more exposure and infection than females (P<0.05). Infected subjects had more exposure (10.2 m(2) min/day) than those not infected (7.44 m(2) min/day) (P<0.05). Compared with uninfected subjects, those infected had 2.9 times more exposure in infected-snail zones (P<0.01). Also, human infection intensity (epg) correlated well with exposure to infected snail zones (r=0.552, P<0.01). People <20 years old had the highest re-infection (21.4%) and intensity (3.77 epg). Median exposure for 20-49-year-olds (9.00 m(2) min/day) was nearly double that of those aged <20 or >50 years old (5.5 m(2) min/day). We conclude that map-referenced water contact and snail evaluation boosts accuracy of activity-diary measurements in large transmission foci for the Asian schistosome. Protecting against faecal contamination of snail inhabited sites, and against occupational exposure for island residents, should be a priority of future research. Potential strategies for migrating buffaloes and families living on visiting fishing boats are explored. PMID- 10838212 TI - Development of an immunoenzymatic assay for the detection of human antibodies against Trypanosoma cruzi calreticulin, an immunodominant antigen. AB - We have developed an indirect immunoenzymatic assay (ELISA) for the detection of human antibodies against calreticulin (formerly known as Tc45), a dimorphic Trypanosoma cruzi antigen, described in our laboratory. PVC microtitration plates were sensitized with the monoclonal anti-calreticulin antibody (MoAb) and reacted with calreticulin present in a partially purified preparation. The presence of anti-T. cruzi calreticulin IgG in sera from infected individuals was tested. The data generated with this assay were validated by correlation, in a regression analysis, with those obtained by an indirect immunoradiometric assay (IRMA). From the 12 seropositive sera (as defined by a commercial test), eight came out positive and four negative in both assays. The 12 human sera were also analyzed in direct immunometric assays (ELISA and IRMA), where the solid phase was sensitized with a whole parasite extract. The direct ELISA and IRMA correlated positively (P<0.01). Further validation of this ELISA was achieved with an indirect immunofluorescense assay. The high degree of significance obtained when the indirect IRMA and ELISA systems were compared, indicated that the relatively small sample number used (12) was statistically satisfactory for the purposes of this investigation. Thus, the IRMA can be replaced by the ELISA, with advantages mainly derived from the cumbersome manipulation of radioactive wastes. The MoAb used as an antigen capture agent in the ELISA proposed here, recognizes a homologous protein in Trypanosoma rangeli, suggesting that individuals infected with this parasite might have crossreactive antibodies. However, the system retains its diagnostic interest, given the facts that the MoAb does not recognize a homologous protein in Leishmania mexicana, Leishmania donovani, or Crithidia fasciculata. PMID- 10838213 TI - Characterization of Leishmania isolated in Iran: 1. Serotyping with species specific monoclonal antibodies. AB - A total of 156 isolates of Leishmania from patients with cutaneous leishmaniasis and one isolate from gerbil were characterized using standard monoclonal antibodies (mAb) in ELISA and IFA test systems. The geographic distribution of the isolates was 30 isolates in Shiraz, 28 in Kerman and 98 in Tehran. A total of 63, 72, and three Leishmania promastigote isolates preferentially reacted with anti-Leishmania tropica mAb (A11), anti-Leishmania major mAb (T1) and anti Leishmania infantum mAb (D2), respectively. Of the three isolates which reacted with anti-L. infantum mAb, two were dermotropic strains of L. infantum and one was the organism isolated from gerbil. Mixed reactions with the monoclonal antibodies were also observed as six isolates reacted with both anti-L. tropica and anti-L. major mAbs, five reacted with anti-L. tropica and anti-L. infantum mAbs and two with anti-L. major and anti-L. infantum mAbs. The monoclonal antibodies used did not react with five of the isolates. The present investigation has identified the organisms from the known and newly emerged foci of cutaneous leishmaniasis in Iran. PMID- 10838214 TI - Morbidity and mortality in booked women who deliver outside orthodox health facilities in Calabar, Nigeria. AB - Women who booked for antenatal care at the University of Calabar Teaching Hospital (UCTH), Calabar, but delivered outside orthodox health facilities were studied. The aims were to determine the pattern of maternal morbidity and mortality in them and to compare this with the pattern in women who booked and delivered at UCTH. One hundred and eighteen of the defaulters traced (35.1%) had complications compared with 34 (10.1%) of the control. Only 32.2% of these defaulters with complications presented in orthodox health facilities for treatment. The major complications in the study group were: perineal tear (19.0%); primary postpartum haemorrhage (12.5%); and puerperal sepsis (5.4%). These were significantly higher in the study group than in the controls (P<0.001). Maternal mortality ratio of 6.0 per 1000 live births was recorded in the study group, but there was no death in the control. Health education and public enlightenment campaigns emphasising universal antenatal care along with delivery in orthodox health facilities are strongly advocated. PMID- 10838215 TI - Comparative sensitivity of dot-ELISA, PCR and dissection method for the detection of trypanosome infections in tsetse flies (Diptera: glossinidae). AB - A visually read dot-enzyme linked immunosorbent assay (dot-ELISA) developed for the detection of trypanosomes in tsetse flies (Glossina spp.) was evaluated in the laboratory and under field conditions. In the evaluation, the fly dissection method was used as a standard technique and compared to the polymerase chain reaction (PCR). In laboratory studies, 133 and 126 tsetse flies were experimentally infected with different stocks of Trypanosoma brucei and T. congolense, respectively. Twenty-five days after infection, the flies were dissected and tested for the presence of trypanosomes using dot-ELISA and PCR. Dot-ELISA detected 98.4% of T. brucei and 94% of T. congolense infections in tsetse midguts, while PCR detected 97.6% of T. brucei and 96% of T. congolense tsetse midgut samples. For field evaluation of dot-ELISA, 700 tsetse flies were caught and screened for trypanosome infections by dissection. Seven of these (1%) had trypomastigotes in the midgut, 23 (3.3%) in the proboscis and none had trypanosomes in the salivary glands. All the flies with midgut infections also had trypanosomes in their proboscides. Five of the seven flies (71.4%) with midgut infections revealed by dissection, were also positive for T. congolense by the dot-ELISA and PCR techniques. Dot-ELISA detected T. congolense infections in an additional 86 (12.4%) of the 700 flies dissected. Of the 23 infections in the proboscis, 16 were T. vivax. Dot-ELISA detected 13 of the 16 (81%) while PCR detected 15 of 16 (94%) T. vivax infections. No T. brucei infection was detected by any of the methods in all the 700 tsetse flies examined. The results obtained from both the laboratory and field studies indicate that the dot-ELISA and PCR techniques are sensitive and species-specific in revealing trypanosome infections in tsetse flies. While dot-ELISA required a single test to detect T. congolense, several primer pairs were needed for PCR. The potential use of dot-ELISA as a tool for studying the epidemiology of trypanosomosis, while considering its field applicability and relatively lower cost is discussed. PMID- 10838216 TI - Development and sexual maturation of Echinococcus vogeli in an alternative definitive host, Mongolian gerbil (Meriones unguiculatus). AB - An alternative definitive host model for Echinococcus vogeli was developed using Mongolian gerbils. The animals treated with prednisolone tert-butylacetate were orally inoculated with protoscoleces of E. vogeli and the development of worms in the small intestine was observed until 50 days post infection (DPI). The parasites were recovered most frequently from the anterior part of the small intestine. The band formation (the sign of proglottisation) of the worm was observed at 7 DPI, the second proglottid and tests formation at 14 DPI, spermatozoa in seminal receptacle and cleavage of ova in uterus at 21 DPI, the hook formation of oncospheres at 28 DPI and the embryophore formation at 34 DPI. Eggs in the feces were first detected at 35 DPI. The coproantigen detection method developed for E. multilocularis could detect the E. vogeli coproantigen early in the infection, at 7 DPI during the prepatent period. Therefore, this technique could be used for the diagnosis of E. vogeli, replacing ordinary fecal egg examination. PMID- 10838217 TI - Comparative analysis of two different subunits of antigen B from Echinococcus granulosus: gene sequences, expression in Escherichia coli and serological evaluation. AB - Two different Echinococcus granulosus antigen B subunits (AgB8/1 and AgB8/2) were characterized and the structure of the genes encoding these two proteins were compared. DNA sequences were expressed in Escherichia coli and the antigens' diagnostic value was then assessed. The genomic sequence of AgB8/1 has a 92 bp intron in the position corresponding to amino acid 16; the AgB8/2 genomic sequence presents a 68 bp intron in the position corresponding to amino acid 20. Both introns are located between the putative N-terminal hydrophobic sequence and the secreted peptide. A comparison between the AgB8/1 and AgB8/2 nucleotide sequences showed a 53.5% identity among exons and a 50% identity between introns. According to the molecular diversity analysis, the elapsed time since both genes shared a common ancestor would be around 4.2x10(7) years. When the native AgB and the two recombinant antigens (rAgB8/1 and rAgB8/2) were tested in an anti-IgG ELISA, the sensitivity of the native antigen B was 77.41% and its specificity was 81.9%, while rAgB8/1 showed 54.84% of sensitivity and 80.17% of specificity and rAg138/2 had an 83.87% sensitivity and a 98.28% specificity. Statistical analysis confirms that rAgB8/2 has a better performance than rAgB8/1 and native AgB in ELISA. PMID- 10838218 TI - Tegumental changes in 21-day-old Schistosoma mansoni harboured in mice treated with artemether. AB - Alterations in the tegument of 21-day-old Schistosoma mansoni, caused by artemether administered to the infected mice, were studied using scanning electron microscopy (SEM). Mice were infected with S. mansoni cercariae, and after 21 days a single dose of artemether (400 mg/kg) was administered intragastrically. After 24, 72 h and 7 days groups of three mice were killed and the schistosomules collected by perfusion, fixed and processed routinely, and examined by SEM. After 24 h, all male and female worms examined showed alterations in the tegument, characterised by swelling, vesiculation and fusion of tegumental ridges; peeling, erosion and collapse of damaged tegumental surface, and also destruction of the oral sucker and acetabulum. After 72 h, severe damage to the tegument was seen, usually including extensive peeling, swelling and vesiculation, and host leukocytes were adhered to the damaged surface. Some worms were surrounded by clusters of host leukocytes or had even disintegrated. Seven days after treatment, some schistosomules still showed severe tegumental damage, but in some cases the damage was less than at earlier times, which suggested that those schistosomules that had survived were beginning to recover. The ability of artemether to cause severe damage to the tegument correlates with its high efficacy in killing 21-day-old schistosomules. PMID- 10838219 TI - Plasmodium falciparum: red blood cell binding studies of peptides derived from histidine-rich KAHRP-I, HRP-II and HRP-III proteins. AB - Histidine-rich proteins have been associated with Plasmodium falciparum infected red blood cells (RBC) cytoadherence, and RBC rosetting; these phenomena may cause clogging of the post-capillary venules, this being one of the main causes of severe cerebral malaria. They may also participate in parasite mature stages' evasion of the immune system and their subsequent destruction in the spleen. Non overlapping synthetic peptides, corresponding to entire amino acid sequences reported for the KAHRP-I, HRP-II and HRP-III proteins, were used in RBC binding assays. Peptides with high and low binding activity were recognized. The KAHRP-I protein shows 3 peptides with high binding affinity to RBCs, two of them variable (peptide 6783, sequence 321QNYVHPWSGYSAPYGVPHGA(340) and peptide 6789, sequence 441KKREKSIMEKNHAAKKLTKK(460)) and the other conserved (peptide 6786, sequence 381KSKKHKDHDGEKKKSKKHKD(400)) having affinity constant of around 190 nM and 1000 binding sites per cell. Interestingly, this peptide shares aminoacid sequences with one reported as being recognized by malaria exposed human antibodies. The HRP-I protein also presents one conserved peptide (peptide 6800, sequence 24NNSAFNNNLCSKNAKGLNLN(43)) with high affinity, located in the amino terminal region of the native protein, having 210 nM affinity constant and 6000 receptor sites. The HRP-III protein only contains peptides with low binding activity. Treatment of red blood cells with neuraminidase reduces the binding of the conserved high binding 6786 and 6800 peptides. Anti-glycophorins A, B and C antibodies inhibit the binding of the conserved high binding 6786 and 6800 peptides. Furthermore, the specific determination of glycoproteins by chemioluminescenoe, in SDS/PAGE western blot, suggests that these glycophorins could be the receptor for these high binding peptides. High binding peptides' critical amino acids, involved in RBC binding were determined by means of competition binding assays. PMID- 10838220 TI - Spatial and temporal dynamics of the secretory pathway during differentiation of the Plasmodium yoelii schizont. AB - A specialized complex of apical organelles facilitates Plasmodium merozoite invasion into the erythrocyte. Even though the apical organelles are crucial to the invasion process, relatively little is known about how they function or their biosynthesis during asexual replication. MAEBL is an erythrocyte binding protein located in the rhoptries and on the surface of mature merozoites and is expressed at the beginning of schizogony before the first nuclear division. Therefore, we have characterized MAEBL as a marker for the biosynthetic pathway of the rhoptry apical organelle during the final phase of intraerythrocytic development and as a marker for the nascent rhoptry vesicle in the immature schizont. An extensive proliferation of the endoplasmic reticulum occurred at the onset of schizogony and was seen as a complex but transient tubule array near the parasite surface. Both the rhoptry protein MAEBL and surface protein MSP-1 appeared to be present in this tubular reticular network together with endoplasmic reticulum markers. MAEBL then transits through Golgi bodies positioned near the parasite plasma membrane, directly adjacent to the network. Rhoptry organelle precursors are seen at the three to four nuclei stage of schizont development, remaining near the plasma membrane throughout schizogony. These studies constitute the first direct evidence that proteins of the rhoptry organelles transit through compartments of the 'classical' secretory pathway. PMID- 10838221 TI - Animal propagation and genomic survey of a genotype 1 isolate of Cryptosporidium parvum. AB - Human cryptosporidiosis is attributed to two major Cryptosporidium parvum genotypes of which type 1 appears to be the predominant. Most laboratory investigations however are performed using genotype 2 isolates, the only type which readily infects laboratory animals. So far type 1 has only been identified in humans and primates. A type 1 isolate, obtained from an individual with HIV and cryptosporidiosis, was successfully adapted to propagate in gnotobiotic piglets. Genotypic characterization of oocyst DNA from this isolate using multiple restriction fragment length polymorphisms, a genotype-specific PCR marker, and direct sequence analysis of two polymorphic loci confirmed that this isolate, designated NEMC1, is indeed type 1. No changes in the genetic profile were identified during multiple passages in piglets. In contrast, the time period between infection and onset of fecal oocyst shedding, an indicator of adaptation, decreased with increasing number of passages. Consistent with other type 1 isolates, NEMC1 failed to infect mice. A preliminary survey of the NEMC1 genome covering approximately 2% of the genome and encompassing 200 kb of unique sequence showed an average similarity of approximately 95% between type 1 and 2 sequences. Twenty-four percent of the NEMC1 sequences were homologous to previously determined genotype 2 C. parvum sequences. To our knowledge, this is the first successful serial propagation of genotype 1 in animals, which should facilitate characterization of the unique features of this human pathogen. PMID- 10838222 TI - An Entamoeba histolytica rab-like encoding gene and protein: function and cellular location. AB - We identified here a 576 bp rab-like gene (EhrabB) in Entamoeba histolytica. EhrabB is located 332 bp upstream from the start codon of the Ehcp112 encoding gene, but is transcribed from the complementary strand. The EhrabB open reading frame predicts a 192 amino acid polypeptide (EhRabB) with 40-42% identity to Rab proteins, involved in vesicle docking regulation in endo and exocytic pathways of eukaryotic cells. Transcripts of 0.6 and 0.97 kb were detected by the EhrabB probe in northern blot assays. Using specific antibodies, EhRabB was located in small cytoplasmic vesicles by confocal microscopy. During phagocytosis, EhRabB was initially translocated to the plasma membrane and to the phagocytic mouths. The protein diminished after 10 min phagocytosis, suggesting that EhRabB could be participating in the regulation of the endocytosis process. PMID- 10838223 TI - Molecular cloning and immunological characterization of phosphoglycerate kinase from Clonorchis sinensis. AB - The parasite Clonorchis sinensis was determined to utilize a large amount of external glucose to carry its energy metabolism. Phosphoglycerate kinase (PGK), a glycolytic enzyme, found in many parasites, has been identified as one of the candidate molecules distinguished from human counterparts for vaccine and drug developments. A cDNA clone purified by screening a C. sinensis cDNA library using a heterologous cDNA probe encoded a putative peptide of 415 amino acids with over 60% identities with PGKs from a number of animals. The putative peptides revealed domains corresponding to 12 beta-sheets and inner loops forming a substrate binding cleft of animal PGKs. The gene product was overexpressed in Escherichia coli and showed a PGK-like enzyme activity. A polyclonal antibody raised against the recombinant C. sinensis PGK was specific to native C. sinensis PGK and localized it to the muscular tissue and tegument of the adult flukes. The C. sinensis PGK elicited antibodies in C. sinensis-infected rabbits. Therefore, it is proposed that C. sinensis PGK could be used as an immunoreagent in the serodiagnosis for clonorchiasis. PMID- 10838224 TI - Isolation and characterisation of sex-specific transcripts from Oesophagostomum dentatum by RNA arbitrarily-primed PCR. AB - In light of the lack of molecular data on the sexual differentiation, maturation and interaction of parasitic nematodes of livestock, the present study investigated sex-specific gene expression in the nodule worm, Oesophagostomum dentatum (Strongylida). Using the technique of RNA arbitrarily-primed polymerase chain reaction (RAP-PCR), 31 expressed sequence tags (ESTs) differentially displayed between the sexes were cloned. Northern blot analysis proved ten ESTs to be expressed exclusively in males (adults and fourth-stage larvae), while two were expressed solely in female stages. None of the ESTs were expressed in infective third-stage larvae. Sequence analysis and subsequent database searches revealed two male-specific ESTs to have significant similarity to Caenorhabditis elegans (predicted) proteins, a protein containing an EGF-like cysteine motif and a serine/threonine phosphatase. Another two male-specific ESTs had similarity to non-nematode sequences. The two female-specific ESTs had similarity to vitellogenin-5 and endonuclease III (predicted) from C. elegans. The remaining ESTs had no similarity to any nucleic acid or protein sequences contained in the databases. The isolation and characterisation of sex-specific ESTs from O. dentatum provides a unique opportunity for studying the reproductive biology of parasitic nematodes at the molecular level, with a view toward novel approaches for parasite control. PMID- 10838225 TI - SmMAK16, the Schistosoma mansoni homologue of MAK16 from yeast, targets protein transport to the nucleolus. AB - The SmMAK16 gene from Schistosoma mansoni was cloned by chance when an adult worm cDNA library was probed with antiserum to affinity-purified S. mansoni GSH S transferases. SmMAK16 encodes a hydrophilic protein of 259 amino acids with a molecular mass of 31 kDa. The protein shares 43% sequence identity and 66% similarity to the nuclear protein MAK16 of Saccharomyces cerevisiae that has been implicated both in cell cycle progression and biogenesis of 60S ribosomal subunits. Both proteins display a similar degree of sequence similar to the hypothetical protein CeMAK16 from Caenorhabditis elegans. These proteins share a number of apparent protein motifs, including two nuclear localization signals (NLS), multiple sites for phosphorylation by protein kinase CK2 and four conserved cysteine residues that resemble a zinc binding domain. SmMAK16 mRNA is more highly expressed in adult female worm than males. Recombinant SmMAK16 was phosphorylated by human protein kinase CK2. When chimeric constructs containing SmMAK16 fused the green fluorescent protein (GFP) were transiently transfected into COS-7s cells, the reporter was localized not in nuclei, but exclusively in nucleoli. The yeast and nematode homologues were likewise able to direct nucleolar accumulation of the fluorescent reporter. The high degree of sequence conservation together with the ability to direct nucleolar protein transport supports the hypothesis that MAK16 proteins play a key role in the biogenesis of 60S subunits. PMID- 10838226 TI - Plasmodium falciparum erythrocyte membrane protein 1 is anchored to the actin spectrin junction and knob-associated histidine-rich protein in the erythrocyte skeleton. AB - A distinctive pathological feature of Plasmodium falciparum malaria is the endothelial attachment of erythrocytes infected with mature asexual-stage parasites in microvessels of the major organs. Electron-dense protrusions described as knobs are displayed on the surface of parasitized erythrocytes and act as attachment points in cytoadherence. Parasite-encoded knob-associated histidine-rich protein (KAHRP) is a major component of knobs found on the cytoplasmic side of the host cell membrane. P. falciparum erythrocyte membrane protein 1 (PfEMP1) is a family of parasite-encoded cytoadherence receptors localized to knobs on the surface of parasitized erythrocytes. Despite its high antigenic diversity, PfEMP1 has a remarkably conserved cytoplasmic domain. We demonstrate in this study that the cytoplasmic domain of PfEMP1 (VAR(CD)) binds to host spectrin and actin and to full-length KAHRP in vitro. Apparent dissociation constants determined for VAR(CD)/F-actin and VAR(CD)/KAHRP interactions are 44.9+/-6.4 and 10. 7+/-2.2 nM, respectively. Further, we provide evidence that KAHRP polypeptides self-associate in solution to form structures similar to knobs and show binding of self-associated KAHRP clusters to spectrin actin-protein 4.1 complexes. Findings in this study suggest that PfEMP1 is localized to the knob in P. falciparum-infected erythrocytes by binding to the host spectrin-actin junction and to self-associated KAHRP through its conserved cytoplasmic domain. PMID- 10838228 TI - Heparin can regulate the binding of Plasmodium falciparum circumsporozoite protein. PMID- 10838227 TI - Blasticidin resistance: a new independent marker for stable transfection of Leishmania. PMID- 10838229 TI - Plasmodium vivax reticulocyte binding protein-2 (PvRBP-2) shares structural features with PvRBP-1 and the Plasmodium yoelii 235 kDa rhoptry protein family. PMID- 10838230 TI - Cloning and high level expression in Escherichia coli of an Anisakis simplex tropomyosin isoform. PMID- 10838231 TI - The lumen of the flagellar pocket of Trypanosoma brucei contains both intact and phospholipase C-cleaved GPI anchored proteins. PMID- 10838232 TI - In this issue 45/1 PMID- 10838233 TI - Charles Augustus Leale and the resuscitation of Abraham Lincoln. PMID- 10838234 TI - Randomised controlled trials of staged teaching for basic life support. 1. Skill acquisition at bronze stage. AB - We have investigated a method of teaching community CPR in three stages instead of in a single session. These have been designated bronze, silver, and gold stages. The first involves only opening of the airway and chest compression with back blows for choking, the second adds ventilation in a ratio of compressions to breaths of 50:5, and the third is a conversion to conventional CPR. In a controlled randomised trial of 495 trainees we compared the performance in tests immediately after instruction of those who had received a conventional course and those who had had the simpler bronze level tuition. The tests were based on video recordings of simulated resuscitation scenarios and the readouts from recording manikins. Differences occurred as a direct consequence of ventilation being required in one group and not the other, some variation probably followed from unforeseen minor changes in the way that instruction was given, whilst others may have followed from the greater simplicity in the new method of training. A careful approach was followed by slightly more trainees in the conventional group whilst appreciably more in the bronze group remembered to shout for help (44% vs. 71%). A clear advantage was also seen for bronze level training in terms of those who opened the airway as taught (35% vs. 56%), for checking breathing (66% vs. 88%), and for mentioning the need to phone for an ambulance (21% vs. 32%). Little difference was observed in correct or acceptable hand position between the conventional group who were given detailed guidance and the bronze group who were instructed only to push on the centre of the chest. The biggest differences related to the number of compressions given. The mean delay to first compression was 63 s and 34 s, and the mean duration of pauses between compressions was 16 s and 9 s, respectively. Average performed rates were similar in the two groups, but more in the conventional group compressed too slowly whereas more in the bronze group compressed too rapidly. Observations were made for only three cycles of compression, but extrapolating these to the 8 min often considered a watershed for chances of survival for victims of cardiac arrest, an average of 308 compressions would be expected from those using conventional CPR compared with 675 for those using bronze level CPR. The implications of this difference are discussed. PMID- 10838235 TI - Utstein style reporting of in-hospital paediatric cardiopulmonary resuscitation. AB - STUDY OBJECTIVE: To report paediatric in-hospital cardiac arrest data according to Utstein style and to determine the effectiveness of cardiopulmonary resuscitation (CPR) in hospitalized children. DESIGN: Retrospective 5-year case series. SETTING: Urban, tertiary-care children's hospital. PARTICIPANTS: All patients who sustained cardiopulmonary arrest. RESULTS: Altogether 227 patients experienced a cardiopulmonary arrest during the study period, 109 (48.0%) were declared dead without attempted resuscitation, and CPR was initiated in 118 (52.0%). The incidence of cardiac arrest was 0. 7% of all hospital admissions and 5.5% of PICU admissions; the incidence of CPR attempts was 0.4 and 2.5%, respectively. Most of the CPR attempts (64.4%) took place in the PICU and the most frequent aetiology was cardiovascular (71.2%). The 1-year survival rate was 17.8%. Short duration of external CPR was the best prognostic factor associated with survival. With few exceptions, the Paediatric Utstein Style was found to be applicable for reporting retrospective data from in-hospital cardiac arrests in children. CONCLUSIONS: In-hospital cardiopulmonary resuscitation was shown to be an uncommon event in children; the survival rate was similar to earlier studies. PMID- 10838236 TI - Arterial blood-gases with 500- versus 1000-ml tidal volumes during out-of hospital CPR. AB - The correct tidal volume during cardiopulmonary resuscitation (CPR) is presently debated. While the European Resuscitation Council (ERC) and American Heart Association (AHA) previously recommended a tidal volume of 800-1200 ml, the ERC has recently reduced this to 400-600 ml. In a prospective, randomised study of 17 non-traumatic out-of-hospital cardiac arrest patients intubated and mechanically ventilated 12 min(-1) with 100% oxygen, we have therefore compared arterial blood gases generated with tidal volumes of 500 and 1000 ml. Mean time from cardiac arrest to arrival of the ambulance was 13+/-8 and 14+/-8 min in the two groups, respectively. Arterial blood samples were taken percutaneously 5 and 10-15 min after onset of the mechanical ventilation and analysed instantly. Pa(CO(2)) was significantly higher for a tidal volume of 500 than 1000 ml at both 5 and 10-15 min, 7.48+/-2.23 versus 3.70+/-0.83 kPa (P=0.002) and 7. 45+/-1.19 versus 3.98+/ 1.58 kPa (P<0.001). The pH was lower for 500 than 1000 ml at 10-15 min, 7.01+/ 0.10 versus 7.20+/-0.17 (P=0.034), with a strong trend in the same direction at 5 min (P=0.06). There was adequate oxygenation with no differences in Pa(O(2)) or BE at any time between the two groups, and no significant differences in any blood gas variables between the 5- and 10-15-min samples. We conclude that arterial normocapnia is not achieved with either tidal volume during advanced life support with non-rebreathing ventilation at 12 min(-1). What ventilation volume is required for CO(2) removal and oxygenation during basic life support with mouth-to-mouth ventilation cannot be extrapolated from the present data. In that situation the risk of gastric inflation, regurgitation and aspiration must also be taken into account. PMID- 10838238 TI - An overview of protective action decision-making for a nuclear power plant emergency. AB - Protection of the public in a nuclear power plant emergency requires decision makers to balance the time requirements from two chains of events: the events associated with a radiological release and the events involved with the response to that release. The management of these events is distributed among personnel at the nuclear plant, in the local community, and in state and federal agencies. All of these parties must coordinate their response to the emergency to assure that timely and effective protective response can be made by the risk area population. This article describes the process by which protective action recommendations (PARs) are developed in nuclear power plant emergency exercises and provides recommendations from research on emergency response in other types of natural and technological hazards. PMID- 10838237 TI - Reperfusion injury after focal myocardial ischaemia: polymorphonuclear leukocyte activation and its clinical implications. AB - The only way to rescue ischaemic tissue is to re-instate the oxygen supply to the tissue. However reperfusion of the ischaemic area not only oxygenates the tissue but also initiates a cascade of processes, which may in some cases result in temporary dysfunction of the myocardium. In order to devise protective measures, it is essential to understand the mechanisms and the triggers of this reperfusion phenomenon. In this review we will mainly focus on the inflammatory response caused by reperfusion. We will cover the different steps of polymorphonuclear leukocyte activation and will briefly discuss the molecular biology of the receptors involved. The currently used pharmacological medications in acute cardiology will be reviewed and in particular their actions on polymorphonuclear leukocyte activation, adhesion and degranulation. This review is a compilation of the current knowledge in the field and the therapeutic progress in the prevention of reperfusion injury made today. PMID- 10838239 TI - Selection of protective action guides for nuclear incidents. EPA. AB - In 1991 the Environmental Protection Agency (EPA) published its revised Manual of Protective Action Guides and Protective Actions for Nuclear Incidents. The protective action guides contained in the manual represent EPA's formal recommendations to Federal, State, and local emergency response officials for protecting public health and safety during a nuclear incident. These guides are expressed in terms of the projected dose at which action(s) should be taken to reduce or eliminate that dose. In determining the appropriate values for the protective action guides, the Agency considered the following four principles: (1) acute health effects should be avoided, (2) the risk of delayed health effects should be minimized, (3) the values should not be higher than justified by a cost-benefit analysis, and (4) the risk to health from implementing the protective action should not be greater than the risk from the dose avoided. This paper examines each of these principles and their application in the selection of the evacuation and sheltering protective action guides for the early, or immediate, phase of a nuclear incident. PMID- 10838240 TI - Protective action recommendations based upon plant conditions. AB - Analyses conducted by the Nuclear Regulatory Commission (NRC) indicate that timely and effective protective action would be necessary to protect the public in a major nuclear power plant accident. Given the large amount of time required to implement an evacuation around most reactor sites, protective action recommendations (PARs) must be based upon specific plant indicators regarding the status of the core and systems that protect the core. This article describes the assumptions made, and the analyses conducted, by the NRC in developing its procedures for PARs based upon plant conditions. PMID- 10838241 TI - Evacuation time estimates for nuclear power plants. AB - Evacuation time estimate (ETE) analyses are conducted to accomplish three objectives. First, they provide data to emergency decision-makers that indicate if evacuation can be implemented in time to significantly reduce radiation exposures. Second, they can be used to determine if ETEs are significantly affected by uncontrollable events such as adverse weather. Third, they indicate whether traffic management actions would significantly reduce evacuation times and provide information relevant to the development of effective traffic management plans. PMID- 10838242 TI - The social psychology of public response to warnings of a nuclear power plant accident. AB - This article reviews the process of public response to warnings of an impending nuclear power plant emergency. Significant evidence exists to suggest that people engage in protective action in response to warnings based upon the substance and course through which emergency warning information is disseminated. The three basic components of a warning system are defined, and the elements of public response to warnings are summarized. Popular myths about public response to warnings are outlined and dispelled based upon current research verification. The conclusion provides an overview and synthesis of the warning response process. PMID- 10838244 TI - Preface PMID- 10838243 TI - Emergency response training: strategies for enhancing real-world performance. AB - Emergency response training poses three problems that are not encountered in training for routine operations. The first of these is a need to remember the provisions of emergency plans and procedures over long periods of time until an emergency occurs. The second problem is a need to generalize from the specific conditions under which training occurred to the potentially very different conditions of an actual emergency. The third problem is a need to develop effective mechanisms for teamwork under conditions that limit retention and generalization. This article identifies nine ways that emergency response training programs can be modified to improve the effectiveness of nuclear power plant personnel who must respond to accident conditions. PMID- 10838245 TI - Nuclear proteins and cell death in inherited neuromuscular disease. AB - X-linked Emery-Dreifuss muscular dystrophy is caused by mutations in emerin, a novel nuclear membrane protein. Other major inherited neuromuscular diseases have now also been shown to involve proteins which localize and function at least partly in the cell nucleus. These include lamin A/C in autosomal dominant Emery Dreifuss muscular dystrophy, SMN in spinal muscular atrophy, SIX5 in myotonic dystrophy, calpain3 in type 2A limb-girdle muscular dystrophy, PABP2 in oculopharyngeal dystrophy, androgen receptor in spinal and bulbar muscular atrophy and the ataxins in hereditary ataxias. This review compares the molecular basis for these various disorders and considers the role of cell death, including apoptosis, in their pathogenesis. PMID- 10838246 TI - Emery-Dreifuss muscular dystrophy - a 40 year retrospective. AB - Emery-Dreifuss muscular dystrophy (EDMD) was delineated as a separate form of muscular dystrophy nearly 40 years ago, based on the distinctive clinical features of early contractures and humero-peroneal weakness, and cardiac conduction defects. The gene, STA at Xq28, for the commoner X-linked EDMD encodes a 34 kD nuclear membrane protein designated 'emerin', and in almost all cases on immunostaining is absent in muscle, skin fibroblasts, leucocytes and even exfoliative buccal cells, and a mosaic pattern in female carriers. The gene, LMNA at 1q21, for the autosomal dominant Emery-Dreifuss muscular dystrophy encodes other nuclear membrane proteins, lamins A/C. The diagnosis (at present) depends on mutation analysis rather than protein immunohistochemistry. It is still not at all clear how defects in these nuclear membrane proteins are related to the phenotype, even less clear that LMNA mutations can also be associated with familial dilated cardiomyopathy with no weakness, and even familial partial lipodystrophy with diabetes mellitus and coronary heart disease! What began as clinical studies in a relatively rare form of dystrophy has progressed to detailed research into the functions of nuclear membrane proteins particularly in regard to various forms of heart disease. PMID- 10838247 TI - Therapeutic screening in the mdx mouse. PMID- 10838248 TI - Pre-clinical screening of drugs using the mdx mouse. AB - The genetically dystrophin-deficient mdx mouse, with its characteristic and regular exercise-induced loss of strength, is a useful experimental platform on which to screen potential drug therapies in the treatment of some dystrophic diseases. Pharmacological agents of several chemical and functional classes were examined in their ability to reduce the loss of muscular strength in young exercised mdx mice. Therapeutic intervention over the period 4-10 weeks of age was evaluated in weekly tests of whole-body strength. This age period represents the most severe manifestation of disease in these animals. Significant improvements in whole-body strength were brought about by treatment with the immunosuppressive and anti-inflammatory drugs prednisone (at low dose only, 1 mg/kg body weight), pentoxifylline (100 mg/kg) and tinset (100 mg/kg). The anabolic hormone insulin-like growth factor-1 (5 mg/kg), as well as the amino acids/metabolites glutamine (10 mg/kg), glutamine plus alanine (each 10 mg/kg), and creatinine (10 mg/kg) all improved strength test performance. The mdx mouse is a responsive system for the screening of potential therapeutic treatments for the muscular dystrophies. PMID- 10838249 TI - A new locus for autosomal recessive limb-girdle muscular dystrophy in a large consanguineous Tunisian family maps to chromosome 19q13.3. AB - Autosomal recessive limb-girdle muscular dystrophies represent a genetically heterogeneous group of diseases characterized by a progressive involvement of skeletal muscles. They show a wide spectrum of clinical courses, varying from very mild to severe. Eight loci responsible for autosomal recessive limb-girdle muscular dystrophies have been mapped and six defective genes identified. In this study, we report the clinical data, muscle biopsy findings and results of genetic linkage analysis in a large consanguineous Tunisian family with 13 individuals suffering from autosomal recessive limb-girdle muscular dystrophy. Clinical features include variable age of onset, proximal limb muscle weakness and wasting predominantly affecting the pelvic girdle, and variable course between siblings. CK rate was usually high in younger patients. Muscle biopsy showed dystrophic changes with normal expression of dystrophin and various proteins of the dystrophin-associated protein complex (sarcoglycan sub-units, dystroglycan, and sarcospan). Genetic linkage analysis excluded the known limb-girdle muscular dystrophies loci as well as ten additional candidate genes. A maximum LOD score of 4.36 at θ=0.00 was obtained with marker D19S606, mapping this new form of autosomal recessive limb-girdle muscular dystrophy to chromosome 19q13.3. PMID- 10838250 TI - A homozygous nonsense mutation in delta-sarcoglycan exon 3 in a case of LGMD2F. AB - We present the first Turkish family with delta-sarcoglycanopathy (LGMD2F). A novel truncating mutation (E93X) in exon 3 was identified in the gene. The index case showed a severe course and there was no cardiac involvement. LGMD2F seems to be rare in our population. PMID- 10838251 TI - Calpain3 expression during human cardiogenesis. AB - Transcripts of calpain3, the gene involved in limb girdle muscular dystrophy type 2A, appear in organs other than the skeletal muscle during human development, the first of which being the early embryonic heart. We examined more precisely the spatio-temporal transcription pattern of calpain3 during human cardiogenesis and the appearance of its protein in fetal tissues, and correlated it to titin expression. Different events of the heart's maturation can be recognized: (i) the presence of titin RNA or protein constitute very precocious developmental cardiac markers appearing before the fusion of the two lateral endocardial tubes; (ii) the disappearance of calpain3 RNA from the ventricular compartment later in the embryonic heart. Finally, although calpain3 transcripts are present in the heart, the corresponding protein is not detected elsewhere than in skeletal muscle. PMID- 10838252 TI - A randomized comparative study of two methods for controlling Tendo Achilles contracture in Duchenne muscular dystrophy. AB - A 30-month prospective randomized study of 27 Scandinavian boys with confirmed diagnosis of Duchenne muscular dystrophy was done to compare the effect of passive stretching combined with the use of night splints (group A) or passive stretching (group B) on the evolution of Tendo Achilles contractures. Assessments were based on the methodology of Scott et al. (Muscle Nerve 1982;5:291 301)Analysis of the pattern and mechanism of dropout was done to eliminate bias between the two groups. Logistic regression showed that Tendo Achilles contracture was the most important variable (P=0.0020) for dropout. Methods of statistical analysis for longitudinal data avoiding induced serial correlations were used in the analysis. The expected annual change in Tendo Achilles contracture was found to be 23% less in group A than in group B after equalization for total muscle strength (%MRC). PMID- 10838253 TI - Minicore myopathy in children: a clinical and histopathological study of 19 cases. AB - Minicore myopathy is a congenital myopathy characterized by multifocal areas of degeneration in muscle fibres. Genetic heterogeneity expected on the basis of clinical variability awaits further resolution. We reviewed 19 cases in order to further delineate the phenotype. Marked hypotonia was the predominant presenting feature, with evidence of antenatal onset in 30% of cases. Weakness was most pronounced axially and proximally, often more severely affecting the shoulder girdle. Mild facial involvement was frequent. Varying degrees of scoliosis were obvious in all patients older than 10 years. In addition, two patients who were also the most severely affected had complete external ophthalmoplegia. One patient showed marked distal involvement. Respiratory failure developed in half of all patients after 10 years of age and correlated strongly with the degree of scoliosis. Cardiac involvement occurred mainly secondary to respiratory impairment. The course appeared static in most cases. Loss of independent walking was observed only in one case at the age of 10 years. On ultrasound scan, differential involvement within the quadriceps was documented in several patients. Variability in fibre size, type 1 predominance and atrophy with occasional type 2 hypertrophy were prominent but nonspecific histological changes. Apart from typical minicores, a marked increase in internal nuclei was the most prominent histological feature. With the exception of one family in which two generations were affected, inheritance appeared autosomal-recessive or sporadic in all cases. PMID- 10838254 TI - Copies of the survival motor neuron gene in spinal muscular atrophy: the more, the better. PMID- 10838255 TI - Regenerative capacity and the number of satellite cells in soleus muscles of normal and mdx mice. AB - Satellite cells are potential myogenic cells that participate in repair and growth of muscle fibres. In this investigation, the change in the number of satellite cells following severe muscle damage was monitored in soleus muscle of age-matched mdx and C57Bl/10 mice. Satellite cells were identified immunohistochemically in the light microscope by their association with a recently described marker protein, M-cadherin, and their location between the muscle fibre's sarcolemma and the surrounding basal lamina. In cross-sections of untreated soleus muscle of C57Bl/10 mice at 11-14. 5 months of age, nuclei of M cadherin positive satellite cells on average amounted to 3.4% of the total number of myonuclei. Surprisingly, significantly higher numbers of satellite cell nuclei, both in absolute numbers (mean 24+/-11 versus 40+/-11 satellite cells per section) and relative to the total number of myonuclei (5. 3%), were found in similarly aged animals in which severe muscle damage had been inflicted 3-6 months before. Cross-sectional area, muscle tissue area and myonuclei counts had recovered to control values. In untreated muscles of age-matched mdx mice satellite cell counts were not different (2.7% of myonuclei) from C57Bl/10 mice. However, regeneration showed marked deficits, as there was a loss of about 36% total cross-sectional area, about 48% total muscle fibre area and about 43% myonuclei per section compared to the untreated mdx muscles. Furthermore, the absolute number of satellite cells decreased from 20+/-11 to 12+/-8 per section. The relative number of satellite cell nuclei remained comparable to, but did not exceed, the undamaged muscles. The poor recovery of muscle and the missing post regeneration rise in satellite cell numbers may indicate the reproductive limits of the satellite pool. PMID- 10838256 TI - Modulation of disease severity in mice with targeted disruption of the acid alpha glucosidase gene. AB - Glycogen storage disease type II (GSDII) is a recessively inherited disorder caused by defects in lysosomal acid alpha-glucosidase. In an attempt to reproduce the range of clinical manifestations of the human illness we have created null alleles at the acid alpha-glucosidase locus (GAA) with several gene targeting strategies. In each knockout strain, enzyme activity was completely abolished and glycogen accumulated at indistinguishable rates. The phenotypes, however, differed strikingly. Acid alpha-glucosidase deficiency on a 129xC57BL/6 background resulted in a severe phenotype with progressive cardiomyopathy and profound muscle wasting similar to that in patients with glycogen storage disease type II. On a 129/C57BL/6xFVB background, homozygous mutants developed a milder phenotype with a later age of onset. Females were more affected than males irrespective of genetic background. As in humans with glycogen storage disease type II, therefore, other genetic loci affect the phenotypic expression of a single gene mutation. PMID- 10838257 TI - High resolution magnetic resonance imaging of the brain in the dy/dy mouse with merosin-deficient congenital muscular dystrophy. AB - Magnetic resonance imaging (MRI) abnormalities in the cerebral white matter are a consistent feature of merosin-deficient human congenital muscular dystrophy, a disease caused by a primary defect in the expression of the laminin alpha2 chain of merosin. To investigate the relationship between imaging changes and merosin deficiency we undertook a MRI study in the dy/dy mouse, an animal model for this form of human congenital muscular dystrophy. High resolution in vivo imaging was performed on anaesthetized animals (two homozygous dy/dy mutants and two heterozygous dy/DY controls, aged 2.5 months) in a dedicated 11.7T magnetic resonance imaging scanner. T(1) and T(2) weighted images were normal in all mice and white matter changes were not seen at a stage of maturity when MRI changes are already very striking in human patients. Cerebral MRI abnormalities do not appear to be a feature of dy/dy mice, despite the virtual absence of merosin expression in the dy/dy mouse brain. Possible causes for this absence of MRI changes, and implications for the pathogenesis of the MRI changes in humans are reviewed. PMID- 10838258 TI - Report of the 70th ENMC International Workshop: nemaline myopathy, 11-13 June 1999, Naarden, The Netherlands. PMID- 10838259 TI - 'An artefact gone awry': identification of the first case of nemaline myopathy by Dr R.D.K. Reye. AB - In 1963, Shy et al. (Brain 1963;86:793-810) and Conen et al. (Can Med Assoc J 1963;89:983-986) published the first description of a novel myopathy characterized by the aggregation of rods (nemaline bodies) in the muscle fibres. This disorder was subsequently known as nemaline myopathy. Dr Douglas Reye, an Australian pathologist, described a patient with 'rod myopathy' five years earlier, in 1958. Here we present Dr Reye's original description of nemaline myopathy, and details of the 'second opinion' which concluded that the rod were a 'processing artifact', so that the case was never published. Detailed histological and immunocytochemical studies of this original case demonstrate the typical features of nemaline myopathy, and a mutation in skeletal muscle alpha actin has recently been identified in this patient. Not only was Dr Reye the first to use the term 'rod' in relation to muscle disease, he also made observations that are relevant to the pathogenesis of nemaline myopathy. PMID- 10838260 TI - 75th European Neuromuscular Centre International Workshop: 2nd workshop on the treatment of muscular dystrophy 10-12 December, 1999, Naarden, The Netherlands. PMID- 10838261 TI - Nutritional support of the cancer patient: issues and dilemmas. AB - Malnutrition in cancer patients results from multifactorial events and is associated with an alteration of quality of life and a reduced survival. A simple nutritional assessment program and early counselling by a dietitian are essential to guide nutritional support and to alert the physician to the need for enteral (EN) or parenteral nutrition (PN). A daily intake of 20-35 kcal/kg, with a balanced contribution of glucose and lipids, and of 0.2-0.35 g nitrogen/kg is recommended both for EN and PN, with an adequate provision of electrolytes, trace elements and vitamins. EN, always preferable for patients with an intact digestive tract, and PN are both safe and effective methods of administering nutrients. The general results in clinical practice suggest no tumor growth during nutritional support. The indiscriminate use of conventional EN and PN is not indicated in well-nourished cancer patients or in patients with mild malnutrition. EN or PN is not clinically efficacious for patients treated with chemotherapy or radiotherapy, unless there are prolonged periods of GI toxicity, as in the case of bone marrow transplant patients. Severely malnourished cancer patients undergoing major visceral surgery may benefit from perioperative nutritional support, preferably via enteral access. Nutritional support in palliative care should be based on the potential risks and benefits of EN and PN, and on the patient's and family's wishes. Research is currently directed toward the impact of nutritional pharmacology on the clinical outcome of cancer patients. Glutamine-supplemented PN is probably beneficial in bone marrow transplant patients. Immune diets are likely to reduce the rate of infectious complications and the length of hospital stay after GI surgery. Further studies are needed to determine the efficacy of such novel approaches in specific populations of cancer patients, and should also address the question of the overall cost-benefit ratio of nutritional pharmacology, and the effect of nutritional support on length and quality of life. PMID- 10838262 TI - Mini-invasive surgery may be advised in oncology. AB - In the last decade mini-invasive surgery has consistently developed with good results, but also with some unjustified clinical applications. This review is aimed at defining evidence based indications and procedures ('clinical practice') and those still worthy of controlled studies in oncologic centers with expertise in mininvasive surgery ('clinical research'). At present, diagnostic and staging laparoscopy and thoracoscopy represent the 'standard' for different tumors. Conversely, therapeutic indications according to evidence based medicine criteria are still limited. Tumors treatment by mini-invasive surgery requires 'expertise' on the part of the surgical team; this can be achieved by extensive training of a correct use of instruments and methods following the general surgical principles of traditional 'open surgery'. PMID- 10838263 TI - Gemcitabine in locally advanced and/or metastatic bladder cancer. AB - Gemcitabine is a promising new drug in patients with locally advanced and/or metastatic transitional cell carcinoma of the urothelium. The drug has been tested as a single-agent in one phase I study and four phase II studies. Gemcitabine was administered on days 1, 8 and 15 every 28 days with a dose in the phase II studies ranging from 1000 to 1250 mg/m(2). Response rates for single agent gemcitabine in as well previously untreated as cisplatin-based pretreated patients ranged from 23 to 29% with CR rates between 4 and 13%. Toxicities were mild to modest and generally without grade 4 toxicities. The combination of gemcitabine and cisplatin has been tested in three phase II studies. Gemcitabine was administered in a dose of 1000 mg/m(2) on days 1, 8 and 15 every 28 days whereas the cisplatin dose and schedule varied. In one study, cisplatin was given in a dose of 35 mg/m(2) on days 1, 8, and 15 together with gemcitabine; in the two other studies in a dose of 70-75 mg/m(2) on day 1 or 2 in each treatment course. The response rates ranged from 42 to 66% with CR rates of 18, 21 and 28%. Median survival was reported in two of the studies, 12.5 and 13.2 months, respectively. Toxicities were generally manageable although the weekly schedule of cisplatin resulted in a high degree of grade 3-4 neutropenia and thrombocytopenia. Thus, the schedule has been optimized by use of monthly cisplatin in a dose of 70 to 75 mg/m(2). The two-drug combination of gemcitabine and cisplatin has also been compared with MVAC in a randomized phase III trial. Gemcitabine was administered in a dose of 1000 mg/m(2) on days 1, 8 and 15 and cisplatin in a dose of 70 mg/m(2) on day 2 every 28 days. The study was initiated late in 1996 and the planned recruitment of 400 patients was reached at the end of October 1998. The results are now eagerly awaited. Preliminary results for gemcitabine tested in two- and three-drug combinations with new agents such as paclitaxel have indicated response rates of up to 79% and these combinations should be further explored. PMID- 10838264 TI - Function of fibroblast growth factors and vascular endothelial growth factors and their receptors in angiogenesis. AB - Angiogenesis, formation of new vessels from pre-existing ones, results from stimulation of endothelial cells, which line the vessel wall. These cells will leave their resting state and start to digest the basement membrane, proliferate, migrate and eventually differentiate to form a hollow tube. All these steps can be induced by growth factors and this review will focus on two important types of angiogenic growth factors, vascular endothelial growth factor (VEGF; also denoted vascular permeability factor, VPF) and fibroblast growth factor (FGF). Both types of factors bind to cell surface expressed receptors, which are ligand stimulatable tyrosine kinases. Binding of the growth factors to their receptors leads to activation of the intrinsic tyrosine kinase and signal transduction to downstream signalling cascades. This results in transcriptional changes and biological responses. The molecular aspects of signalling cascades critical for endothelial cell proliferation and migration are beginning to be delineated. In contrast, signalling cascades leading to endothelial cell differentiation remain to be determined. Angiogenesis is essential for a number of physiological events such as embryonic development, ovulation, and wound healing. It has become increasingly clear that a number of diseases depend on angiogenesis. For future development of therapeutic tools, it is important to understand the molecular mechanisms that regulate angiogenesis. PMID- 10838265 TI - Radiotherapy in head and neck cancer in the elderly: a challenge. AB - Elderly patients represent the most rapidly growing subgroup of the patient population in France and in the majority of industrialized countries. The effect of age in terms of the prognosis and response to treatment remains unclear. The management strategy (curative versus palliative) for head and neck cancer in the elderly has given vent to divergent opinions and controversies in several respects (the type and quality of treatment, quality of life and economic consequences). This review only focuses on the radiotherapy schedule and head and neck cancers. We compare aged patients with head and neck cancer to younger patients in terms of clinical features, tumor biology, type of treatment, side effects and response. We conclude that if the patient is in a good general condition following a complete evaluation of the cancer, physicians should propose curative treatment with radiotherapy because retrospective trials demonstrate that response in older patients when treated aggressively is comparable to that of younger patients. However, specific trials concerning aged patients with head and neck cancer, quality of life and radiotherapy are warranted. PMID- 10838266 TI - Asymptomatic bacteriuria in human immunodeficiency (HIV)-infected women. AB - Objective: To determine the prevalence of bacteriuria in HIV- infected women versus healthy non-HIV-infected controls.Methods: A prospective cohort study was undertaken between January 1996 and March 1997. One hundred nineteen ambulatory, premenopausal women from the Women's HIV Clinic and 191 similar non-infected Gynecology Clinic controls were recruited. Information recorded from the medical record and patient questionnaire included: age, prior history of UTI, antibiotic usage, sexual frequency, diaphragm/spermicide use, and CD4 count. A clean catch urine culture was collected on all patients. Statistical analysis was via SPSSPC.Results: The mean age of HIV-infected women was 33 vs 26 for non-infected. The mean CD4 count was 341. A prior history of UTI (mean # per lifetime) was significantly higher for HIV women (2.8) vs controls (1.8) [P =.01]. The mean weekly sexual frequency was significantly higher in HIV-negative (2.1) vs HIV infected (1.2) [P <.001]. Diaphragm/spermicidal use was uncommon but equal in both groups. The prevalence of any growth of uropathogens (>10(2) cfu/mL) was 30% in the HIV-infected vs 47% in controls. The percentage of asymptomatic bacteriuria for each range of uropathogen titer (cfu/mL) in urine was:Chronic antibiotic usage (39% of HIV-infected vs 4% of controls) was associated with a decreased prevalence of bacteriuria, but there was not a significant difference in prevalence of bacteriuria between groups. Regardless of titer ranges of uropathogens evaluated, the prevalence of bacteriuria was not different between CD4 counts of <200 and >/=200. As compared to controls, HIV-infected individuals demonstrated shifts toward nosocomial pathogens: Enterococcus, S. aureus and non E. coli gram-negative rods.Conclusion: The prevalence of bacteriuria was not greater in HIV-infected women versus non-HIV-infected women versus HIV non infected controls and was not significantly affected by antibiotic usage or CD4 count. Uropathogens responsible for asymptomatic bacteriuria in HIV-infected women are different from "healthy" controls and are more representative of nosocomial flora. PMID- 10838267 TI - Yield of postoperative fever evaluation. AB - Introduction: In women undergoing major gynecologic surgeries, we wish to determine the frequency and yield of blood cultures, urine cultures, and chest x ray as part of postoperative fever evaluation.Methods: A retrospective review of 537 consecutive patients undergoing major gynecologic surgery. Postoperative fever was any temperature >/=38 degrees C. In patients who developed postoperative fever, it was determined if blood cultures, urine cultures, and chest x-rays were performed and how frequently these yielded positive results.Results: Two hundred eleven patients (39%) developed postoperative fever. Blood cultures were obtained in 77 of 211 (37%) febrile patients, urine cultures in 106 of 211 (50%) febrile patients, and chest x-rays in 54 of 211 (26%) febrile patients. Zero of 77 blood cultures were positive, 11 of 106 (10%) urine culture were positive, and 5 of 54 (9%) chest x-rays were positive. Logistic regression revealed that late onset fever predicted for positive urine cultures and early onset fever and advanced age predicted for pneumonia. Eighty percent of patients with pneumonia were symptomatic.Conclusion: Although postoperative fevers are routinely evaluated by blood cultures, urine cultures, and chest x-rays, they rarely yield positive results. PMID- 10838268 TI - A prospective, randomized, clinical study to compare the clinical safety, effectiveness, and cost of oral ofloxacin/clindamycin vs intravenous clindamycin/gentamicin for the treatment of postpartum endomyometritis. AB - Objective: The primary objective of this prospective, randomized, clinical study was to compare the safety, clinical and microbiologic efficacy, and cost of oral ofloxacin in combination with clindamycin vs intravenous (IV) clindamycin/gentamicin in the early empiric treatment for hospitalized patients with mild to moderate postpartum endomyometritis. The secondary objective is to reduce total hospital and patient treatment cost. Postpartum endomyometritis is a major cause of infectious morbidity in the obstetric patient. It is the most common complication associated with cesarean delivery. Careful timing and amniotomy, limited vaginal examinations, and prophylactic antibiotics for cesarean section delivery may help to reduce the incidence and severity of endomyometritis. Endomyometritis is caused by bacteria that compose the normal cervicovaginal flora. These are anaerobic gram-positive cocci (Peptostreptococcus and Peptococcus), aerobic streptococci (Group B Streptococci and enterococci), Enterobacteriaceae, Bacteroides (B. fragilis, B. bivius, and B. disiens), and clostridium species.Ofloxacin is a synthetic broad-spectrum antibacterial agent for intravenous and oral administration. Following oral administration, the bioavailability in tablet form is 98% with maximum serum concentrations in 1 to 2 hours. Steady state concentrations are achieved after 4 doses. Ofloxacin usually is bactericidal in action. A synthetic broad-spectrum antibacterial agent for intravenous and oral administration. Ofloxacin inhibits DNA topoisomerase (ATP hydrolyzing), commonly referred to as DNA-gyrase. DNA-gyrase causes double stranded DNA breakage; it inhibits duplication, transcription, and repair of bacterial DNA.Methods: This is a preliminary study that has enrolled 19 evaluable patients towards the overall enrollment of 60 patients for statistical significance. Patients clinically diagnosed as having postpartum endomyometritis who meet the inclusion/exclusion criteria were entered into the trial. Patients were examined for the presence of fever (102.2 degrees F), pelvic pain, and foul lochia. A medical history, physical examination, and laboratory analysis were obtained prior to the first dose of antibiotic treatment. A signed consent was obtained prior to the study enrollment and randomization. Appropriate endometrial, blood, and urine culture specimens were obtained prior to the initiation of antibiotic therapy.Patients in Group 1 were treated with oral therapy using ofloxacin 400 mg q12h plus clindamycin 900 mg q8h until 24 hrs of afebrility. In Group 2, patients were treated with clindamycin 900 mg IV q8h plus gentamicin IV 5mg/kg/d q 8h until afebrility. Antibiotic therapy was continued for at least 48 hours unless significant clinical deterioration occurred necessitating the withdrawal of the patient from the study.Results:Conclusions: We found in our preliminary study that oral ofloxacin in combination with oral clindamycin was equally as efficacious, well tolerated, and safe as the combination of intravenous therapy with clindamycin and gentamicin for the treatment of postpartum endomyometritis. PMID- 10838269 TI - Simplified cesarean section: a strategic surgical approach to minimize postoperative infectious morbidity. AB - Objective: A simplified method of cesarean delivery aimed at minimizing postoperative morbidity is illustrated.Methods: Two hundred consecutive cesarean deliveries were performed by the authors' simplified cesarean technique. Mean patient age was 27 years (range 17-46), and mean weight was 169 pounds (range 112 414). Indications for cesarean delivery included dystocia or failure to progress in labor (38%), repeat cesarean (32%), malpresentation (11.5%), fetal distress (9.5%), and other (9%).Results: Simplified cesarean delivery was successfully completed in all cases. Mean operating time was 16 minutes (range 9-33), mean blood loss was 460 mL (range 100-1150), and mean postsurgical hospitalization time was 72 hours (range 36-120). No bowel, bladder, or vascular injuries occurred. Postoperative febrile morbidity occurred in one patient (0.5%), ileus occurred in one patient (0.5%), and blood transfusion was administered to one patient (0.5%). No cases of wound infection, wound dehiscence, hematoma, or incisional hernia occurred. All patients were ambulatory on the first postoperative day. All but one patient (99.5%) tolerated a regular diet on the first postoperative day.Conclusions: The authors' technique of cesarean section appears to be a safe and efficient method for cesarean delivery associated with minimal postoperative infectious morbidity and rapid resumption of bowel and ambulatory function. PMID- 10838270 TI - Single-dose antibiotic prophylaxis during cesarean section. AB - Objective: To compare single-dose antibiotic prophylaxis (cefotetan 1 g vs cefoxitin 2 g) in various subpopulations based upon risk factors for postsurgical infection following cesarean section.Methods: Patients undergoing cesarean section from April 1993 through March 1994 were included in a retrospective analysis if either of the above antibiotics were administered, surgery was non emergent, gestational age was less than 32 weeks, absence of fever or prior antibiotics therapy within 72 hours, and no history of organ transplantation or HIV. Cases classified as high risk for infection: IDDM, obesity, autoimmune disease, sickle cell disease, or corticosteroid use. Cases classified as high risk for endometritis (any 2 factors): labor >12 hours, >4 vaginal examinations, ruptured membranes >9 hours, and internal fetal monitor. Cases were separated into 4 groups: elective vs non-elective, low vs high surgical risk. A chi(2) analysis was used to test for differences in infection rates between groups (P <.05).Results: Of 1383 cesarean sections, 385 met criteria for inclusion. Non elective cases accounted for 77% of cases. Postsurgical infection rate was greater in non-elective cases, 7.4%, vs elective cases, 3.0% (P =.056) as was the rate of endometritis (3.2% vs 1.2%, P =.185). No differences were noted based on antibiotic regimen. Postsurgical infection rate was greater for 28 cases at high risk for both surgical infection and endometritis (17.9%) when compared to all 357 other cases (4.5%), P =.003. No difference was noted for endometritis. Of the 28 cases 28.6% of patients treated with cefoxitin and 7.1% of cases treated with cefotetan developed postsurgical infection (P =.13).Conclusion: Overall cefoxitin and cefotetan provided equivalent clinical outcome. A small subset of patients with multiple risk factors for infection may benefit from cefotetan. PMID- 10838271 TI - Chorioamnionitis: the association between clinical and histological diagnosis. AB - Objective: To determine the association between clinical and histological diagnoses of chorioamnionitis.Methods: A retrospective 9-year review of hospital records for all perinatal deaths (N = 804) occurring at a Western New York regional perinatal center between 1988 and 1996 was conducted. Information concerning clinical indicators of chorioamnionitis were recorded including: maternal fever, leukocytosis, uterine tenderness, foul smelling discharge, and fetal tachycardia. Histological examination of the placenta was conducted in 797 of 804 perinatal deaths and staged by a standardized method. Type and number of clinical signs and symptoms were correlated with severity of histologic grade to determine their relation.Results: Chorioamnionitis was diagnosed either clinically or histologically in 243 (30.5%) of 797 perinatal deaths. Of these, 72 cases (23.5%) occurred in the 307 fetal deaths and 171 (34.9%) occurred in the 490 neonatal deaths. Five cases were diagnosed only on autopsy. In 51 of 243 cases (21%) there was histologic confirmation of chorioamnionitis without any clinical indicators present. In 13 of 243 cases (5.4%) there were 1 or more clinical indicators of chorioamnionitis without histologic confirmation. Sensitivity and specificity for 1 or more clinical indicators was 77.8% and 97.7%, respectively.Conclusion: Chorioamnionitis was present in 30.5% of perinatal deaths in this cohort. Identification of one or more clinical signs or symptoms was found to be both a sensitive and specific screening method for a presumptive diagnosis of chorioamnionitis when using histologic confirmation as the gold standard. PMID- 10838272 TI - Febrile morbidity and hospital stay in high-risk cesarian section patients at a non-teaching hospital. AB - Objective: The objective of the study was to determine the incidence of postcesarian febrile morbidity and relate this to hospital stay in a high-risk indigent population treated at a private non-teaching hospital.Methods: This was a retrospective chart review of all patients done between January 1995 and December 1996. Discharge summaries, antepartums, progress notes, and labs were reviewed for each patient. Of 257 charts reviewed, 5 were inadequate for various reasons. Board-certified surgeons performed and assisted in the operations. Twenty-one patients had scant prenatal care and 6 had no prenatal care. All patients had the abdomen scrubbed with Betadine soap prior to painting. No shaving was done. Gloves were changed after closure of uterine incision. The pelvis was copiously irrigated with 3-4 L of saline. The subcutaneous layer was irrigated from a height of 6-12" with 12 to 1 L of fluid. After this step, this layer is not touched by anything from the operating field.Results: Of 162 patients with primary cesarian, 20 had postoperative fever, 18 with endometritis, 2 with wound infections. All but 5 of these patients had labor as ruptured membranes of 12 hours or more. Four had prolonged 2nd stage. Of 28 failed VBACs, 2 had fever vs none for 59 elective repeat cesarians. The average hospital stay for febrile patients was 4.4 days vs 2.7 for afebrile patients. The incidence of wound infection was 0.8%. The incidence of fever was 12.2% for primary cesarians and 8.8% in the total study group of 249 patients.Conclusion: This study demonstrates that the adoption of simple measures presented in Methods can dramatically decrease the incidence and severity of postcesarian fever, especially wound infection, thereby allowing safe, early hospital discharge. PMID- 10838273 TI - Strategies to decrease costs associated with GBS prophylaxis in preterm gestations. AB - Objective: 1) To evaluate the costs associated with a practice of routinely culturing women for Group B beta-Streptococcus (GBS) who are at risk for preterm delivery. 2) To evaluate the charges associated with a policy of empiric antibiotic therapy in women suspected of imminent delivery prior to 37 weeks gestation.Methods: In our hospital, women who present with preterm contractions (PTC), preterm labor (defined as uterine contractions resulting in cervical change, PTL), and preterm rupture of the membranes (PROM) were cultured for GBS. Women with PTL and PROM received parenteral antibiotic therapy pending availability of culture results (48 hours). We reviewed the records of women who participated in a study to determine the sensitivity and specificity of a rapid test for GBS (Am J Obstet Gynecol 1997;176:S35). Results of the Lim broth culture, our gold standard, were used for this analysis. The charges for performing cultures ($32/culture) and administering intravenous ampicillin ($54/dose) were calculated.Results: There were 118 women with PTC, 52 women with PTL, and 64 with PROM. Thirty-four (14.5%) of the cultures were positive. The results are presented below.In women with PTC, the incidence of GBS was 12.7%. The culture results were available and guided care in only the 6 women who delivered between 2 and 7 days. In women with PTL the incidence of positive cultures was 11.5%. All of the women received antibiotics pending results of cultures. The majority of the women had delivery delayed 48 hours until results were available. Care was modified in the 7 women who delivered between 2 and 7 days. In women with PROM the incidence of GBS was 20.3%. The culture results were available and guided therapy in 11 women.Conclusions: Our findings suggest that GBS cultures have limited clinical value in women with PTC. Only 6 women, 5.5%, had their care modified as a result of the cultures, at a cost of $3776. In women with PTL, we over-treated the majority of women with antibiotics, most of whom delivered after culture results were available. Cultures had minimal impact on management. Antibiotics should be reserved for women whose labor is progressing on tocolysis. In women with PROM, most delivered prior to availability of culture results. We would have saved $2048 had no cultures been obtained. These results are probably irrelevant, if routine antibiotic administration is utilized in PROM to prolong latency intervals regardless of GBS status. PMID- 10838274 TI - Rubella and pregnancy: failed prevention policies. AB - Objective: Postpartum rubella immunization can prevent 33-50% of all cases of congenital rubella. Our purpose was to assess missed opportunities for postpartum rubella vaccination.Methods: A retrospective prenatal record review of 304 women attending a high risk obstetrics clinic was done. The rubella status on the first prenatal visit was recorded. Rubella status was determined by immunoassay (ELISA) and reported as immune, uncertain, or nonimmune. For rubella nonimmune women their ethnicity, age, parity, and place of delivery were recorded.Results: Fifteen (4.9%) of the 304 women were nonimmune to rubella. Of the 15 women, 6 were white, 4 were Afro-American, and 5 were Hispanic. The mean age was 30.1 +/- 6.2 y (range 20-42 y). Thirteen had given birth at least once (range 1-4), while two were nulliparous. Eleven of the 13 women gave birth in the USA, while two gave birth in South America. These eleven women gave birth 20 times without receiving rubella counseling or vaccination.Conclusion: Recommendation for postpartum rubella vaccination of nonimmune women should be adhered to in order to decrease the risk of congenital rubella syndrome. PMID- 10838275 TI - Potential for routine concurrent determination of chlamydia and cervical abnormalities by single fluid-based sampling. AB - Objective: The FDA recently approved adjunctive HPV testing from the vial of PreservCyt(R) Solution used for the ThinPrep(R) Pap Test. We have now evaluated the potential for routine chlamydia testing from cellular material collected for primary Pap testing using the fluid-based ThinPrep method.Methods: Cervical scrapings were collected for a conventional Pap smear and residual material adhering to the sampling implement after slide preparation was suspended in PreservCyt Solution. Chlamydia testing was performed on this residual material using two methods. 1) A slide was prepared using the ThinPrep 2000 System for a determination by direct fluorescence. 2) One mL of the cell suspension was removed and processed for determination by an amplified DNA probe technique. A separate conventional endocervical swab sampling from the same patient was used for testing by an independent direct DNA probe technique (reference method). One hundred specimens were selected and intentionally biased, 19 positive and 81 negative based on the independent swab results, for comparison to the slide and amplified DNA probe methods.Results: The slide-based method gave a sensitivity of 84% (16/19) and a specificity of 100% (81/81). Of 3 discrepant specimens, 2 tested positive by the amplified probe method, while all 3 remained negative by repeat DFA. The amplified probe method gave a sensitivity of 95% (18/19) and a specificity of 99% (80/81) after analysis of discrepant samples by repeat testing.Conclusions: Feasibility has been demonstrated for accurate and effective routine testing for chlamydia with either DFA or DNA probe methods, from a single, fluid-based sample already approved for primary Pap screening. PMID- 10838276 TI - The association of asymptomatic bacterial vaginosis with upper genital tract infection. AB - Objective: To determine whether asymptomatic bacterial vaginosis (BV) is associated with upper genital tract inflammation or bacterial colonization.Methods: Fifty nonpregnant women with intact uteri who planned to undergo gynecologic surgery and had no recent symptoms of vaginal infection were enrolled. We obtained a vaginal swab for Gram stain, endocervical swab for chlamydia and gonorrhea DNA probe testing, and Pipelle endometrial biopsy for aerobic and anaerobic cultures and histology. We correlated surgical findings and histology of available surgical specimens with the microbiologic results. The diagnosis of BV was made according to Speigel's criteria. Bacteria isolated from the uterus were classified as high virulence versus low virulence. Contingency tables were analyzed using the chi-square or Fisher Exact tests.Results: Twenty one of 50 patients had BV on Gram stain, 3 had intermediate BV, and 3 had unreadable slides. Eleven patients had histologic endometritis, including 7 of the 21 with BV. The association between BV and endometritis was not statistically significant. Thirty-eight patients had a positive endometrial culture; 18 of these also had BV and 3 had intermediate BV. Of patients with a positive endometrial culture, isolates from those who also had BV or intermediate BV were more likely to be highly virulent pathogens compared to those who tested negative for BV (P <.05). No patients had positive gonorrhea or chlamydia tests. Of the 37 patients who underwent abdominal surgery, 20 had visible adhesions. The association between adhesions and endometritis, BV, or positive endometrial cultures was not significant.Conclusion: Patients with asymptomatic BV are more likely than patients without BV to have highly virulent bacteria isolated from the endometrium. BV may predispose to upper genital tract infection. PMID- 10838277 TI - Safety and efficacy comparison of two dosing regimens of MetroGel-Vaginal(R) in the treatment of bacterial vaginosis: a multicenter, randomized, single-blind, parallel comparison. AB - Objective: To compare the safety and efficacy of 0.75% metronidazole vaginal gel (MGV) administered QD for 5 days to MGV administered BID for 5 days for the treatment of bacterial vaginosis (BV).Methods: Nonpregnant women 18 years or older with a diagnosis of BV were enrolled into this trial. Patients were determined to have BV if their vaginal discharge had >/=20% clue cells and if they had at least 2 of the following criteria: homogeneous discharge, positive KOH amine odor test, or vaginal pH >/=4.7. In addition, microbiologic confirmation of BV at baseline was required to be evaluable. Patients were evaluated at baseline and at 7-12 days after completion of therapy. If patients remained free of BV after therapy, they were evaluated at a follow-up visit 28-35 days after therapy.Results: A total of 514 patients were enrolled in the trial; 252 in the QD treatment group and 262 in the BID treatment group. Clinical cure rates based on patients considered evaluable at first return visits were 77% and 80% for QD and BID dosing regimens, respectively. Clinical cure rates based on patients considered evaluable at the final visit were 58% and 61% for QD and BID dosing regimens, respectively. Clinical cure rates at the final return visit based on intent-to-treat analysis were 53% and 57% for the QD and BID regimens, respectively. Two adverse events in two patients were reported as serious. Both adverse events resolved and neither event was judged to be related to therapy.Conclusion: MetroGel-Vaginal administered QD for 5 days was found to be as effective as MetroGel-Vaginal administered BID for 5 days in the treatment of bacterial vaginosis 28-35 days after therapy. The number of patients reporting adverse events judged to be related to therapy was also statistically comparable in both treatment groups. PMID- 10838278 TI - Can a multimedia educational computer-based program on genital herpes teach patients about their disease? AB - Objective: Education of patients with genital herpes about their disease is time consuming. To evaluate the effectiveness of an educational computer program, we developed a multimedia interactive presentation to teach patients about genital herpes. Such programs can supplement clinician visits for patients with genital herpes, or those at risk for HSV acquisition.Methods: Patients seeking care for genital herpes, or those at risk for HSV acquisition, were asked to participate in the program during routine clinic visits at 5 physician's offices nationwide. A self-administered 7 item herpes knowledge questionnaire was given before and after participation. An additional questionnaire evaluating the satisfaction with the program was also self-administered at completion.Results: 428 patients were enrolled and completed the pre- and post-knowledge questionnaire and 332 patients completed the satisfaction survey. On the pre-test, 20.1% of patients answered all questions correctly, 65.4% answered correctly 4 to 6 questions, and 14.5% 3 or less. On the post-test, 32.9% of patients answered all questions correctly, 61.5% answered correctly 4 to 6 questions, and 5.6% 3 or less (P <.001 for pre- and post-test comparison). A positive change in knowledge between pre- and post test was seen on 6 of 7 items (P <.001 for all 6). The overall satisfaction with the program was high: the mean rating was 6.2 on a scale 1 (poor) to 7 (excellent).Conclusion: Computer-based education programs about genital herpes may provide a useful adjunct to teaching in physician offices and result in knowledge gain about the disease, at least short-term. Such programs may assist in management of chronic sexually transmitted infections. PMID- 10838279 TI - Percutaneous penetration of Aldara cream, 5% during the topical treatment of genital and perianal warts. AB - Objective: The objective of this single-center, open-label trial was to evaluate the percutaneous penetration of Aldara (imiquimod) cream, 5% when applied topically to patients with anogenital warts using a more frequent/aggressive dosing regimen.Methods: Ten otherwise healthy males and six otherwise healthy, nonpregnant, nonlactating females with histology results suggestive of, or diagnostic of, human papilloma virus/condyloma acuminata were enrolled. Females were required to be practicing an acceptable form of contraception control. Patients applied cream daily (8 +/- 2 hours) until complete wart clearance, or for a maximum of 16 weeks. Following the initial dose, at approximately week 4, and at the end-of-treatment, patients were confined for 42 hours in order to obtain a series of blood and urine samples. These samples were analyzed for levels of imiquimod and two metabolites, S-26704 and S-27700. Biological marker levels were not included as a part of this trial.Results: No quantifiable (>/=5 ng/mL) levels of imiquimod or the two metabolites were observed in any of the serum samples collected. Five patients had quantifiable (>/=10 ng/mL) imiquimod, S-26704, or both in urine. No quantifiable levels of S-27700 were observed. Complete clearance of warts occurred in 40% of male patients and 83% of female patients. Erythema was the most frequently observed local skin reaction and was moderate in intensity, although 6 of 16 patients reported a severe erythema reaction at some point in the study. Application site reactions (itching, burning and pain) were the most frequently reported adverse events.Conclusion: The lack of quantifiable levels of imiquimod or metabolites in serum, together with sporadically occurring quantifiable but low levels in urine, indicate that systemic exposure, after daily application of Aldara cream to genital/perianal skin, may occur but is minimal; however, pharmacological (immune marker) effects were not evaluated because cytokine measurements were not obtained. A future trial assessing cytokine levels after topical Aldara therapy with minimal systemic levels of imiquimod would help assess systemic drug and pharmacological effects and utility of this product in pregnant women. PMID- 10838280 TI - Mechanism of action of imiquimod 5% cream in the treatment of anogenital warts. AB - Objective: The objective of this randomized, double-blind, placebo-controlled trial was to evaluate the mechanism of action of imiquimod cream in the treatment of anogenital warts, and to apply the findings to the results of previously conducted safety and efficacy trials.Methods: Imiquimod (16 patients) or placebo (3 patients) cream was applied 3 times a week for up to 16 weeks; cream remained on the skin overnight for 8 +/- 2 hours. Wart biopsies were taken at prestudy, week 6, and the end of treatment (just prior to clearance or at week 16) and analyzed using PCR for HPV/DNA and RT-PCR for mRNA to identify cytokines, cellular markers, markers of proliferation and differentiation, and viral gene products. Efficacy was assessed based on wart area regression as documented by wart area measurements and photographs.Results: All patients enrolled in the trial had HPV type 6/11. All imiquimod-treated patients experienced a >/=75% clearance in baseline/target wart area. Imiquimod treatment stimulated significant increases in mRNA for IFN-alpha, 2'5' AS and IFN-gamma. Increases in mRNA for CD4, CD8, and TNF-alpha were also observed, suggesting activation of a T helper type-1 cell mediated response. During the trial one of the vehicle treated patients also experienced spontaneous wart clearance; comparisons of the cytokine levels for this patient were similar to those observed for the imiquimod treated patients.Conclusions: The results of this mechanism of action trial indicate that the stimulation of local cytokines and cellular infiltrates by imiquimod leads to a reduction of HPV types 6 and 11 viral load with subsequent wart regression and normalization of keratinocyte proliferation without evidence of scarring. In two previous randomized vehicle-controlled trials evaluating patients with anogenital warts, the majority of patients had HPV-DNA types 6 or 11 as assessed by in situ hybridization. These results provide additional insight into the mechanism of total clearance for these otherwise healthy patients. The Th1 response demonstrated in this trial also explains the lower total clearance rates demonstrated in HIV-positive and AIDS patients. PMID- 10838281 TI - Assessment of vulvovaginal complaints: agreement between phone and office management. AB - Objective: To examine the agreement between nursing phone and provider office management of vulvovaginitis.Methods: Patients who called the Kaiser Permanente Phone Call Center with vulvovaginal complaints were evaluated by a registered nurse, who took a standardized history, and made an assessment and plan; additionally, she made a judgment regarding whether she would have treated the patient over the phone. These patients were seen in the office the same day and examined and treated by a clinician following a protocol that included cultures for typical vaginal and cervical pathogens. Providers also made a judgment about phone treatment. Nurses and providers were blinded to each others' diagnosis and therapy. Kappa coefficients were used to evaluate the interexaminer agreement between nurses on the phone and providers in the office with respect to diagnosis and treatment.Results: Four hundred eighty-five patients underwent phone evaluation. Of these, 253 (52%) completed the study protocol and were considered in the final analysis. Kappa values for the diagnosis of trichomoniasis (0.05), bacterial vaginosis (0.12), candidiasis (0.22), chlamydia, herpes simplex, and urinary tract infections (0.05), and "other" (0.05) demonstrated poor agreement between nurses and providers. There was also poor agreement between phone nurses and providers regarding the necessity of an office visit (0.14).Conclusion: This prospective study challenges the notion that the telephone is an effective tool for diagnosing and treating vulvovaginal complaints. Patients with these complaints should be evaluated in the office. Alternatively, programs should be developed and analyzed to educate nurses involved in telephone triage of vulvovaginitis. PMID- 10838282 TI - Vulvodynia: in situ hybridization analysis for human papillomavirus. AB - Objective: To determine the incidence of human papillomavirus (HPV) DNA by in situ hybridization analysis in women with vulvodynia and suspected HPV infection as compared to normal controls.Methods: A chart review was performed to identify women with vulvodynia and both colposcopic and histologic evidence of HPV infection. Control patients of women with vulvar biopsies for benign, non-HPV related conditions were also identified from the chart review. Vulvar biopsies from 11 women with vulvodynia and 12 control patients were compared by histology and by in situ hybridization for HPV DNA. In situ hybridization was performed with DNA probes for common HPV subtypes: 6,11,16,18,30,31,33,34,45,51, and 52. Positive controls were performed for each HPV subtype.Results: All 11 patients with vulvodynia and both clinical and histologic evidence of HPV infection were negative for all types of HPV DNA by in situ hybridization. The 12 control patients who had no clinical or histologic evidence of active HPV were also negative for HPV DNA by in situ hybridization.Conclusions: DNA from the tested HPV subtypes was not identified in patients with vulvodynia and suspected HPV infection. While it is possible that other HPV subtypes may be involved in vulvodynia, these data suggest that DNA from common HPV subtypes is not identified in greater frequency in patients with vulvodynia as compared to normal controls and suggest that the role of HPV in vulvodynia may be overestimated. PMID- 10838283 TI - The semi-quantitative assessment of menstrual flow in relation to clinical and hematological parameters. AB - The hemostasis of menstruation has not been extensively studied particularly since the development of laboratory tests for von Willebrand factor, a glycoprotein that plays a critical role platelet aggregation.Objective: To determine the relationship of menstrual flow to clinical and hematological parameters.Methods: We studied 155 (56 blood type O/99 blood type non-O) normal female volunteers that met the following criteria: age 12-50; not taking an oral contraceptive, estrogen, or Coumadin; not pregnant; regular periods. A questionnaire was administered regarding the patient's medical history. A pictorial chart score of menstrual blood flow was obtained using standard tampons/pads for all participants; this semi-objective measure has previously been validated to be diagnostic of menorrhagia at a score of >185. Menstrual coagulation sampling was within the first 3 days of the cycle. We then analyzed by multiple linear regression analysis the pictorial chart score assuming a log normal distribution as a dependent variable with the following clinical and laboratory parameters as independent variables.Results: The mean age was 35.7 +/- 7.4 years. The mean pictorial chart score was 119.1 +/- 76.3.Conclusion: A history of prior births and laboratory evidence of a low von Willebrand factor antigen appear to predict for increased menstrual flow; studies are ongoing in applying these parameters in women presenting with menorrhagia. PMID- 10838284 TI - The substantial impact of von Willebrand disease on menstruation. AB - von Willebrand disease (vWd) is the most common of all congenital bleeding disorders with an estimated prevalence of 1-3% in the general population. However, the gynecological complications have not been thoroughly described.Objective: To compare the clinical and quality of life aspects of vWd in menstruating women in relation to a cohort of menstruating women in the general population.Methods: A patient questionnaire and provider survey of the medical and quality of life aspects of menstruation was administered to 81 menstruating vWd patients registered at four geographically linked Hemophilia Treatment Centers. The questionnaire was also administered to 150 menstruating women volunteers that comprised a control group used to determine normal coagulation levels in menstruating women. We assessed the impact on quality of life of menses in both of the cohorts by a Likert scale of 1-10 with 10 being "most significant impact" using 7 quality of life parameters with those comparisons by Wilcoxon rank sum test.Results: 88% of the vWd patients (pts) had type I vWd, the remaining Type II or unknown. The mean age of the vWd patients was 31.6 +/- 10.3; the mean age of the control group was 35.5 +/- 7.6. The following comparisons were made using chi(2) and Wilcoxon rank sum test:Conclusions: vWd markedly diminishes the quality of life during menses. This observation warrants efforts to reduce the attendant morbidities of vWd in menstruating women. PMID- 10838285 TI - Use of adenosine for pharmacological cardioversion of SVT in pregnancy. AB - Objective: To evaluate the success and safety of adenosine use in treating maternal supraventricular tachycardia (SVT) in pregnancy.Methods: Hemodynamically stable pregnant women diagnosed with supraventricular tachycardia admitted to Jackson Memorial hospital from 1990 to 1995 and treated with adenosine were included in the study. Adenosine was given intravenously with an initial dose of 6 mg, followed by two 12 mg doses as needed. Mothers and fetuses were followed prospectively to assess short-term and long-term effects of the treatment.Results: A total of four pregnant patients were treated with adenosine for medical cardioversion of SVT. All patients were hemodynamically stable upon arrival to the maternity ward and were successfully treated with adenosine. Both mothers and fetuses were monitored during treatment. Adenosine successfully converted the SVT to sinus rhythm, in all four patients. One patient was treated successfully twice, at 15 weeks and subsequently at 24 weeks. No complications occurred to either the mothers or the fetuses during the treatment. Five and ten minute APGAR scores at birth were greater than seven. Mothers and babies were followed from 1 month to 5 years after treatment and no contributory deleterious effects have been found. One patient was lost to follow-up.Conclusion: Our four patient cases, added to the limited published cases in the literature, support adenosine's safety and efficacy for the medical treatment of supraventricular tachycardia in the hemodynamically stable pregnant woman. PMID- 10838286 TI - The effects of cigarette smoking on pulmonary function tests during pregnancy. AB - Objective: The objective of this study was to determine what changes occur in pulmonary function testing in pregnant women who smoke versus nonsmokers.Methods: This was a prospective study. Pregnant women were recruited in all trimesters of pregnancy. A total of 34 women (15 smokers, 19 nonsmokers) were recruited. Measurements of forced vital capacity (FVC), forced expiratory volume in one second (FEVI), the ratio of FEVI to FVC, and maximum voluntary ventilation (MVV) were obtained.Results:Conclusions: This study confirms other studies that have seen no significant changes in FVC, FEVI, FEVI/FVC in pregnant smokers and nonsmokers. A significant worsening in MVV was seen in pregnant women who smoke. The pathophysiologic impact of this effect is unclear. However, since MVV is a measure of the bellows action of the chest, this may lead to a decrease in oxygenation during the active phase and the second stage of labor. This may lead to more abnormal fetal monitoring strips and possibly lower Apgar scores. Further investigation will be necessary to clarify the effects on pregnancy of lower MVV in patients who smoke. PMID- 10838287 TI - Normative oxygen saturation values for pregnant women at sea level. AB - Objective: To develop normative ranges of maternal arterial oxygen saturation (SpO(2)) for women residing at sea level throughout gestation and postpartum.Materials and Methods: Healthy, nonsmoking, non-laboring pregnant women, with no obstetrical risk factors, were recruited into this longitudinal study performed at The Stamford Hospital (elevation sea level). Maternal SpO(2) was measured in the dominant index finger while breathing room air with a Nelcor N-200 pulse oximeter and DS-100A sensor (Nellcor Puritan Bennett, Pleasanton, CA) at set antepartum (AP) and postpartum (PP) intervals. Oxygen saturation was measured in the following positions: sitting, lying on the left then right side, knee-chest, and supine. Five minutes were allowed between measurements. Measurements were taken at 12, 24, and 36 weeks of gestation and then at 12 weeks postpartum for each patient.Results: Thirty-six patients were initially enrolled. Three were excluded from this study, secondary to loss of follow-up (1), preterm delivery at 35 weeks (1), and complicated placenta previa (1). Of the 33 patients who remained in the study 5 have delivered and undergone postpartum evaluation. SpO(2) values (%) are summarized as mean +/- 2 SD in the Table.Conclusion: Preliminary data in this ongoing descriptive longitudinal study suggest that maternal SpO(2) is at least 97% throughout pregnancy for healthy, non-smoking women residing at sea level. These normative data may be useful in managing pregnant women at risk for hypoxia. PMID- 10838288 TI - Hemodynamic and hormonal changes during pneumoperitoneum and trendelenburg positioning for operative gynecologic laparoscopy surgery. AB - Objective: The number and types of surgical procedures being preformed using laparoscopic technique is increasing due to technological advances. Recent studies suggest the carbon dioxide pneumoperitoneum and patient positioning causes hemodynamic alterations, respiratory acidosis, and a release of stress hormones. However, to date, no studies have investigated the physiological effect of laparoscopic procedures lasting more than 60 minutes on the stress response and the effect of Trendelenburg positioning. The purpose of this study was to identify the physiological effect of pneumoperitoneum and positioning during prolonged laparoscopy on hemodynamic (cardiac index, mean arterial pressure, heart rate, systemic vascular resistance, and stroke volume), metabolic (arterial blood gases), and hormone (arginine vasopressin, aldosterone, and plasma renin activity) parameters. We hypothesized that pneumoperitoneum and patient positioning will alter the hemodynamic, hormone, and metabolic parameters.Methods: The study was longitudinal in design and sampled a total of 31 healthy subjects having a gynecologic oncologic laparoscopic procedure at Hurley Medical Center, Flint, Michigan. The subjects were randomly assigned one of three groups receiving an initial insufflation pressure of either 10, 15, or 18 mmHg. After obtaining informed consent hemodynamic, metabolic, and hormone measurements were obtained at the following times: 1) pre-induction, 2) post induction, 3) post-insufflation, 4) post-Trendelenburg 5 minutes and at 30, 60, 90, and 120 minutes post-insufflation. The results were analyzed using multivariate analysis of variance for repeated measures with a P <.05. A power of 0.9 was obtained to identify changes over time.Results: During the time course of the study the hemodynamic, metabolic, and hormonal parameters showed significant alterations. The most dramatic hemodynamic changes occurred post-insufflation characterized by a decrease in cardiac index and stroke volume with a concurrent increase in systemic vascular resistance. The metabolic parameters showed a significant decrease in pH and corresponding increase in PaCO(2). However, the pH and PaCO(2) remained within normal limits through the study. As part of the study's protocol the investigators increased minute volume to control for a rise in PaCO(2) during the procedure. A significant increase was noted in aldosterone and arginine vasopressin at post-insufflation and Trendelenburg positioning. Plasma renin activity showed a dramatic increase following post-insufflation. None of the subjects developed any post anesthetic complications.Conclusion: Our study demonstrated that pneumoperitoneum and Trendelenburg positioning cause statistically significant elevations in the stress hormones and concurrently cause a decrease in hemodynamic parameters. A healthy patient may tolerate these changes but a patient with cardiovascular disease or pulmonary problems may not be able to compensate as efficiently. PMID- 10838289 TI - Incidence, clinical characteristics, and timing of objectively diagnosed venous thromboembolism during pregnancy. AB - Objective: To determine the incidence, timing, and associated clinical characteristics of objectively diagnosed pregnancy-associated venous thromboembolism (VTE).Methods: A retrospective review of VTE cases occurring between 1978 and 1996 was performed. Cases of deep venous thrombosis (DVT) and pulmonary embolism (PE) were identified by ICD-9 discharge diagnosis code and review of antepartum and coagulation laboratory databases. Study inclusion criteria required the objective diagnosis of VTE with either Doppler ultrasound, impedance plethysmography, pulmonary angiography, ventilation-perfusion scanning, or CT/MRI.Results: Among 268,525 deliveries there were 165 (0.06%) episodes of VTE (1/1627 births). There were 127 cases of DVT and 38 cases of PE. Only 14% (23/165) had a prior history of DVT or PE. Most DVTs occurred in the left leg (104/127, 81.9%). Nearly three quarters of the DVTs (95/127, 74.8%) occurred in the antepartum period. Among the antepartum DVT cases, half were detected prior to 15 weeks of gestation (47/95, 49.5%), with only 28 cases occurring after 20 weeks (P <.0001). The majority of the PEs occurred in the postpartum period (23/38, 60.5%). There were only 3 maternal deaths due to PE, all associated with cesarean section. Only 1 patient developed PE while on heparin therapy for DVT while 11 others had complications attributable to heparin use.Conclusion: Most pregnancy-related VTE occurs in the antepartum period. The risk of deep venous thrombosis appears to begin early in pregnancy, even before the second trimester. The highest risk period for pulmonary embolism is after cesarean delivery. Maternal complications of heparin anticoagulation during pregnancy are rare. PMID- 10838290 TI - Effect of oral contraceptives containing 20 and 35 ug ethinyl estradiol on urinary prostacyclin and thromboxane levels in smokers and non-smokers. AB - Objective: Several observational epidemiological studies have shown an increased risk of myocardial infarction (MI) in oral contraceptive (OC) users who smoke cigarettes. Nearly a quarter of all OC users are smokers. The etiology of MI in OC users has been shown to be thrombotic, and the thrombogenic effect of OCs appears to be due to the estrogen component of the formulation. There are epidemiologic data that indicate that decreasing the estrogen dosage in OCs decreases the risk of thrombosis. Smoking may enhance thrombosis in OC users by decreasing prostacyclin levels. The objective of the present study was to investigate the interaction between smoking and OC use with respect to thrombogenesis by studying the effects of OCs and smoking on urinary levels of prostacyclin PGI and thromboxane A(2) TX in smokers and non-smokers.Study Design: A total of 60 healthy women, aged 19-32 years, who were not taking any hormonal treatment for at least 3 months prior to initiating the study were divided into three equal groups: A) OC users who smoked (N = 20), B) OC users who did not smoke (N = 20), and a control group of 10 smokers and 10 non-smokers. To be eligible to participate in the study, all smokers had to smoke at least one pack of cigarettes per day. Each OC treatment group was randomized to receive either norethindrone (NET) acetate (1 mg)/ethinyl estradiol (EE(2)) (0.035 mg) (N = 10) or NET acetate (1 mg)/EE(2) (0.02 mg) (N = 10) daily for 3 months. Overnight urine collections and fasting blood samples were obtained at baseline and at 3 months prior to taking the last OC. Serum levels of NET and EE(2), as well as urinary levels of cotinine and the stable metabolites of prostacyclin and thromboxane A(2) (6-keto-prostaglandin F(1alpha) and thromboxane B(2), respectively), were measured by specific immunoassays.Results: Study subjects showed compliance with respect to smoking and OC use as determined by the levels of serum NET, EE(2), and cotinine. Comparing mean levels of PGI metabolites in the two groups of OC smokers, those taking 35 ug EE(2) showed a significant decrease in PGI. Comparing the ratio of PGI/TX in all smokers, only the smokers on the 35 ug formulation showed a significant decrease in the ratio, while there was no change in the 20 ug group. No significant changes in the ratio PGI/TX during the same time interval in smoking and non-smoking controls and OC non smokers were observed.Conclusions: Women smokers taking 20 ug EE(2) OC had no thrombophilic changes of PGI and TXA(2) metabolites. Women smokers taking 35ug EE(2) oral contraceptive had thrombophilic changes in PGI/TXA metabolite ratio. Women smokers who use oral contraceptives may have less thrombotic risk taking formulations with 20ug EE(2) than 35ug EE(2). PMID- 10838291 TI - New reference levels for CA125 in pre- and postmenopausal women. AB - Objective: CA125 is a glycoprotein commonly produced by the endometrium. Serum CA125 is widely useful as a marker for ovarian cancer screening, with 35 U/mL the generally accepted upper limit of normal. Since CA125 is known to fluctuate with vaginal bleeding and menopausal status, the purpose of this study was to develop different reference ranges for normal women of differing physiologic states.Methods: We retrospectively analyzed 1,407 patients with serum CA125 levels taken at Cleveland Clinic Florida between 1992 and 1997. Patients with obvious elevation of CA125 from gynecologic cancers were excluded. Student t test was used to determine statistical significance.Results: Among 1,407 patients studied, 256 were premenopausal, 46 perimenopausal, 957 postmenopausal, and 148 excluded due to known gynecologic cancers. There was a significant difference between the mean CA125 values of premenopausal (19.3 +/- 15.6) and postmenopausal women (11.7 +/- 9.2) with P.7 x 10(-6). Among premenopausal women, there was a statistically significant difference between CA125 levels during menses (21.4 +/- 19.3) and luteal phase (14.0 +/- 9.1) with P =.03. Among the subgroup of postmenopausal women with known bleeding history, the mean CA125 values for women with vaginal bleeding was 12.49 +/- 11.5 compared to those without bleeding, 9.62 +/- 4.6 (P =.017).Conclusions: For normal premenopausal women, the overall upper limit of CA125 should be 50 U/mL. However, if menstrual status is known, the upper normals should be: 62 during menses, 51 for proliferative phase, and 32 for luteal phase. For postmenopausal women, the CA125 levels should be no more than 35 for those with vaginal bleeding and 20 for those without bleeding. The stratified CA125 values may help provide more specific ovarian cancer screening. PMID- 10838292 TI - Management of Krukenberg tumors: an 11-year experience and review of the literature. AB - Purposes: The Montefiore Medical Center (MMC) experience with Krukenberg tumors (gastrointestinal cancer metastatic to the ovary) was reviewed 1) in order to determine whether it could be used to make clinical management recommendations, eg, differences in treatment based on menopausal status, secondary debulking, prophylactic oophorectomy; 2) to compare the MMC experience to the reported literature, since one of our patients survived 1912 months beyond the generally accepted mean of this rare tumor.Methods: 1) Prospective study from January 1985 April 1996 of 5 patients followed at MMC with a diagnosis of gastrointestinal (GI) cancer metastatic to the ovary; 2) retrospective chart review of all female patients admitted to MMC with a diagnosis of GI cancer during the same time period; 3) computerized literature review from 1966 to 1996.Results: Five patients were followed prospectively. Retrospective chart review identified 1,016 female patients admitted with gastric (231) or colon (785) cancer, 2 additional cases of Krukenberg tumor were identified. Of the 7 patients with Krukenberg tumors the primary tumor was colon in 5 and gastric in 2. The average age at presentation was 49.3 years (range 35-80); 5 were premenopausal, 2 were postmenopausal, and 2 (28.6%) were postpartum. The average survival of these 7 patients was 12.3 months (range 4 days to 26 months). The 5 patients who had ovarian metastases removed at first laparotomy survived an average of 10.8 months. The 1 patient who had a secondary debulking survived longest (26 months). The 1 patient who had no surgery lived 6 months. These numbers are not statistically significant, but do suggest that further study is warranted. Including the prospectively studied patients, 1,021 patients were evaluated. Seven of 1,021 (0.7%) had Krukenberg tumors. Two of 11 (18.2%) premenopausal gastric cancer patients had a Krukenberg tumor, whereas none of the 222 postmenopausal gastric cancer patients did. Similarly, 3 of 41 (7.3%) premenopausal colon cancer patients had a Krukenberg tumor compared to 2 of 747 (0.3%) postmenopausal colon cancer patients. This difference in menopausal status is significant by Fisher's Exact test (P <.001). Nineteen patients underwent oophorectomy at the time of primary colon cancer surgery: 11 had normal or atrophic ovaries, 2 thecomas, 4 simple cysts, and 2 Krukenberg tumors. Three of 19 (15.8%) were premenopausal.Conclusions: Review of the literature has identified a number of diagnostic and management issues that appear to impact on survival. These include timing of definitive diagnosis of Krukenberg tumors, ie, before, after, or at the same time as diagnosis of the GI primary tumor; menopausal status; concurrent pregnancy; role of debulking; prophylactic oophorectomy. The prognosis worsens when the primary tumor is identified after the metastasis to the ovary is discovered. Krukenberg tumor is more common in premenopausal women than in postmenopausal women. The number of patients with Krukenberg tumors discovered postpartum in our study was significantly increased compared to the other series in the literature. The number of patients who received prophylactic oophorectomy or secondary debulking in our study was inadequate to draw conclusions regarding a benefit of these procedures. However, a benefit may be suggested for debulking, as survival appears to be increased. More importantly, there may be a role for prophylactic oophorectomies in both pre and postmenopausal patients, as this would eliminate the need for a repeat laparotomy. A randomized trial is needed to evaluate the role of prophylactic oophorectomy and debulking. Since the clinical and pathologic details in the literature vary widely, it is extremely difficult to compare studies, particularly the treatment and survival of patients with Krukenberg tumors. Reports should include age, site of GI primary, time from diagnosis of primary to ovarian metastasis, and overall survival as well as survival from the time of diagnosis and treatment of the Krukenberg. We wish to alert the clinician that persistent GI symptoms always warrant investigation. Pelvic inflammatory disease, pregnancy, and postpartum endometritis may mask the GI symptoms. Delays in diagnosis should be avoided. During surgery, the gynecologic surgeon must do a complete upper abdominal exploration, and the general surgeon must do a complete pelvic evaluation. Since Krukenberg tumors are rare, a national registry should be started to gather information on these patients; this might lead to better diagnosis and treatment. PMID- 10838293 TI - Gene therapy of epithelial ovarian cancer using adenoviral vectors. AB - Ovarian cancer is the fourth most common cause of death in women. Gene therapy using the herpes simplex virus thymidine kinase (HSV-tk) gene followed by ganciclovir (GCV) treatment has been successfully applied in the treatment of different cancers in experimental animals and in humans. In a recent report, we have demonstrated that the HSV-tk/GCV system can be used efficiently to kill human epithelial ovarian cancer cells (Gynecol Obstet Invest 1997;43:268-75). In this work, we wanted to test the ability of the HSV-tk/GCV to treat ovarian cancer in an animal model.The immune-deficient nude mice model was employed, and mice were injected intraperitoneally with the human epithelial ovarian cancer cell line OVCAR3, 10(8) cell/mouse. The mice were divided into three different groups, groups 1 and 2 were treated by intraperitoneal injection of adenovirus carrying the HSV-tk gene (ad-tk) on day 3 after cell implantation. Group 1 received 2 x 10(8) pfu/mouse; group 2 received 20 x 10(8) pfu/mouse. Group 3 did not receive any viral injection and served as our negative control. All mice received GCV 10 mg/kg IP bid for 6 days. All mice were hosted in the same facilities and had access to food and water ad libitum. Mice in group 3 started to show clinical manifestations of disease by day 10, and all mice were dead by day 21 (16 +/- 1.5). At this point mice in groups 1 and 2 appeared perfectly healthy. Autopsy done on group 3 mice demonstrated multiple cancer implants in the abdominal cavity plus hemorrhagic ascitis. In contrast, autopsy on sample mice from groups 1 and 2 at the same time point failed to demonstrate any macroscopic or microscopic cancer.On further follow-up, mice in groups 1 and 2 started to show cancer-related signs, eg, weight loss, movement difficulty, poor reflex response, and finally death. Survival varied between 50 and 101 days with a mean of 66 +/- 17 days for group 1 and 74 +/- 13 days for group 2. Autopsy done on treated mice demonstrated multiple cancer implants and ascitis. In conclusion, a single injection of ad-tk/GCV was able to improve survival in an ovarian cancer mouse model from an average of 16 days to 74 days. Trials with multiple injection in a novel immune-competent mouse model of ovarian cancer are underway in our laboratory. PMID- 10838294 TI - Factors leading to delay in diagnosis of invasive cervical cancer. AB - Objective: Screening for cervical cancer has been one of the major successes in American health care. The reasons for screening failure have not been well described. We have encountered 24 patients in the past year on our gynecologic oncology service who have presented with grossly visible stage IB or higher cervical cancers. A questionnaire was developed to determine why these patients presented with invasive cervical cancer, a theoretically preventable disease.Methods: All patients seen at our hospital center with grossly visible cervical cancer were offered entry into this prospective study, approved by the IRB. After informed consent was obtained, a questionnaire was administered to the patients by a physician. The questions involved sexual and gynecologic history, symptom review, social history, and personal concerns involving gynecologic exams. Twenty-four patients were approached for the study and 20 were enrolled. Four patients were unable to participate secondary to medical or social conditions that did not allow them to complete the questionnaire.Results: Of the 20 patients enrolled in this study, 14 of 20 (70%) presented with bleeding irregularities. Upon further questioning, 19 of 20 (95%) reported a history of abnormal bleeding. Only 3 of 20 (15%) patients reported a history of an abnormal Papanicolaou smear, and 7 of 20 (35%) reported not having had a Papanicolaou in the past 5 years. Two patients had never had a Papanicolaou smear. Seventeen of 20 (85%) of all patients in the study had medical insurance at the time of diagnosis, and only 2 patients reported lack of insurance as the reason they had not been seen by a gynecologist in the past 2 years. The majority of the study patients, 17 of 20 (85%) had contact with medical personnel in the 2 years prior to diagnosis. Sixteen of the 20 patients had been seen by a physician, 5 of 20 had surgery or other invasive procedures, and 6 of 20 had been seen in an emergency department. Only 5 of 20 patients knew that a Papanicolaou smear is a screening test for cervical cancer. Five of the 20 thought it to be a test for infections, and 10 of 20 did not know the purpose of a Papanicolaou smear. The main reason women gave for not having had a Papanicolaou smear in the past 2 years was that they felt healthy and had no symptoms (12/20).Conclusion: The majority of patients in our study (17/20) reported contact with a physician within 2 years of diagnosis of their invasive cervical cancer. It appears that failure to screen and lack of understanding regarding Papanicolaou smear screening, rather than lack of access to medical care, is a major cause for delay in diagnosis of cervical cancer. PMID- 10838295 TI - Small cell carcinoma of the cervix: a clinical study of 15 patients and review of the literature. AB - Objective: Small cell carcinoma of the cervix is a very rare and aggressive tumor. Most gynecologic oncology centers have little experience with this tumor. The purpose of the study was to evaluate the University of Kansas' experience with this tumor and compare findings with those found in current literature.Methods: Fifteen patients with small cell carcinoma of the cervix were treated at or in association with Kansas University Medical Center between 1977 and 1997. Clinical data including age of patient, pregnancy history, stage of tumor, recurrence, type of therapy, presenting symptoms, location of metastases, and overall survival were studied.Results: Age at diagnosis ranged from 20 to 83 years with a mean of 47. Two patients were nulliparous, 2 were primiparous, and 11 were multiparous. Five patients (33%) were stage I, 3 (20%) were stage II, 1 (7%) was stage III, and 6 (40%) were stage IV at diagnosis. Four patients (27%) had progressive courses without good response to treatment, 7 (47%) recurred at an average of 15 months. The patients were treated with surgery, radiation, chemotherapy, or a combination thereof. Extrapelvic metastases developed in 5 of 8 patients with stage I or stage II disease with distant nodes, liver, lung, and brain being common sites. Three patients (20%) developed brain metastases. Tumor lysis syndrome was encountered in one patient. One patient was alive and well 80 months after diagnosis and one patient was lost to follow-up. The remaining 13 died of their disease. Mean survival was 22.3 months for stage Ib, 40 months for stage II, 27 months for stage IIb, 8 months for stage III, and 19.2 months for stage IV.Conclusions: Small cell carcinoma of the cervix is a rare and aggressive variant of cervical cancer. Our experience with this tumor raises the question of increased incidence of central nervous system metastases with small cell carcinoma. Present therapy has not significantly improved outcome with this tumor. There is conflicting evidence regarding the etiologic role of an infectious agent. Tumor lysis syndrome is a possible risk when treating these patients. PMID- 10838296 TI - Endometrial carcinoma under the age of 40: reappraisal for oophorectomy in stage I disease. AB - Objective: To reappraise the rationale of castration and the necessity of retroperitoneal lymph node biopsy in patients with endometrial carcinoma under the age of 40 years.Methods: A retrospective review of 30 patients under the age of 40 among 815 patients who had primary endometrial cancer treatment was carried out. The review consisted of clinical data, surgical pathology, and survival rates, as well as a comparative study of the literature.Results: The average age was 35.1 years. Three-quarters of the patients (76.6%) had stage I disease. Almost half of them had risk factors for endometrial carcinoma. The pathologic type was adenocarcinoma in 83% and grade 1 in 57%. Among 23 patients with stage I disease, 7 (30.4%) had no residual disease. Even with 16 patients found to have residual disease, 10 had it confined to the endometrium. Ovarian malignancy was only seen in 2 instances. Five demonstrated corpus luteum. Only 3 (13%) had lymph node metastasis and all in advanced disease. Six (20%) of these patients had delayed treatment more than 6 months. The follow-up was 5 months to 19 years. All are living and free of disease in stage I and stage II. Both patients with stage III disease died.Conclusions: In view of low risks in these young women with stage I disease, a thorough surgical staging including lymph node resection is desirable and hysterectomy with ovarian preservation is the treatment of choice. Oophorectomy might be considered in patients with cancer of the ovaries or in advanced stages of disease. PMID- 10838297 TI - Primary malignant melanoma of the uterine cervix: case report and review of the literature. AB - Objective: To present a case of malignant melanoma of the uterine cervix and review the medical literature, evaluate the clinical presentations and prognosis, and establish a plan of management for this rare pathology.Methods: A case of cervical malignant melanoma treated at our institution was prospectively followed until her death. An extensive MEDLINE search was then performed to obtain all previous case reports on this entity. Information regarding age, clinical presentation, staging, pathology, lymphnode status, treatment, and survival was reviewed.Results: Our patient is a 70-year old white female who presented with a 4-month history of irregular vaginal bleeding. She was diagnosed with malignant melanoma of the cervix, stage IIa, for which she underwent a modified radical hysterectomy, partial vaginectomy, and pelvic and para-aortic lymph node dissection. She received adjuvant radiation therapy to the pelvis, total dose of 4500 cGy. The patient's survival was 29 months. There have been only 24 published cases of primary cervical melanoma in the medical literature. The patients' ages ranged from 39 to 78 years old. The main presenting symptom was vaginal bleeding (67%). The majority of the patients, 58%, presented in early stages (Ia to IIa). Treatment was varied, ranging from a simple excision of a mass to radical hysterectomy with lymph node dissection and adjuvant radiation or chemotherapy. Survival was widespread, even within the same stage.Conclusion: Cervical melanoma is a rare disease of which no prospective or retrospective studies exist, only case reports. Treatment controversies exist. We recommend radical surgical excision to obtain negative margins and lymphadenectomy only for grossly positive nodes. The prognosis is poor and unpredictable. PMID- 10838298 TI - A randomized prospective study of the use of endometrial ablation for prevention of recurrent endometrial polyps in breast cancer patients receiving tamoxifen. AB - Objective: To assess the roles of endometrial ablation in prevention of recurrence of tamoxifen-associated endometrial polyps in breast cancer patients.Design: A randomized prospective study of tamoxifen-treated patients who underwent hysteroscopic removal of endometrial polyps with or without simultaneous resection of the endometrium.Materials and Methods: Twenty consecutive women (aged 43-61 years) undergoing hysteroscopic removal of tamoxifen-associated endometrial polyps were randomized via a computer-generated random table to undergo or not to undergo concomitant endometrial ablation. All patients had undergone endometrial sampling prior to the procedure. The patients were followed for at least 18 months (range 18-24 months). The follow-up included transvaginal ultrasonography every 6 months and hysteroscopy when endometrial irregularity was noted. The main outcome variable was recurrence of endometrial polyps while the occurrence of uterine bleeding was noted.Results: In the 10 study group women, who underwent endometrial ablation, only 1 patient had a 1 x 1 cm endometrial polyp diagnosed and removed during the follow-up period. Seven of the study women remained amenorrheic, and 3 experienced spotting a few days every month. In the control group, a recurrent endometrial polyp, necessitating hysteroscopic removal, was diagnosed postoperatively in 6 women (two-tailed Fisher's Exact test; P <.06).Conclusion: Recurrence of endometrial polyps, one of the most common problems in breast cancer patients receiving long-term treatment with tamoxifen, can be significantly reduced by performing endometrial ablation at the time of hysteroscopic removal of the polyp. The possible risk of occult endometrial cancer is yet to be determined. PMID- 10838299 TI - Positive margins after cervical conization as an indicator of residual dysplasia. AB - Objective: To determine the significance of positive margins of resection after cervical conization as an indicator of residual dysplasia.Materials and Methods: A retrospective analysis for patients who underwent cervical conization either by loop electrosurgical excision procedure or cold knife conization between 1986 and 1997 at Montefiore Medical Center and North Central Bronx Hospital. The factors evaluated included grade of dysplasia with respect to positive or negative margins and post-procedure follow-up. Differences among groups were evaluated using the chi(2) and Fisher's Exact test.Results: Of a total of 179 patients, 99 had positive margins of resection and 80 had negative margins of resection. Twenty-six patients had CIN I, of which 10 had positive margins of resection and 16 had negative margins of resection. There were 30 patients with CIN II, of which 13 had positive margins and 17 had negative margins. One hundred fourteen patients had CIN III/carcinoma in situ (CIS), of which 68 had positive margins of resection while 46 had negative margins of resection. Nine patients had microinvasive disease (MIC), of which 8 had positive margins of resection and 1 had negative margin of resection. The correlation between higher grades of dysplasia and the likelihood of having positive margins was noted to be statistically significant (P =.02). Patients were followed up from a period of 6 weeks to 5 years.Of the patients with CIN I and positive margins, 5 had a normal post-cone Papanicolaou smear while 2 had an abnormal post-cone Papanicolaou smear. Seven of 9 patients with CIN II and positive margins had normal initial post-procedure Papanicolaou smear while only 2 had abnormal initial post follow up Papanicolaou smear. Twenty-five patients with CIN III/CIS and positive margins had normal Papanicolaou smears at their initial post-procedure follow-up while 36 patients had an abnormal initial follow-up. Six of 8 patients with MIC and positive margins had documented follow-up. Of these 6, 2 had normal post procedure Papanicolaou smears while 4 had abnormal post-procedure Papanicolaou smears. This approaches statistical significance. Additionally, the incidence of residual disease was analyzed in hysterectomy specimens with respect to grade of dysplasia. No patients with CIN I and positive margins were treated with hysterectomy. Of the 3 patients with CIN II treated with hysterectomy, 2 had residual dysplasia. Of the 21 patients with CIN II/CIS who underwent hysterectomy, 10 had residual disease. Of the 5 patients with MIC who underwent hysterectomy, 3 had residual disease.Conclusion: The likelihood of positive margins of resection increases with higher dysplasia. The incidence of abnormal initial post-procedure Papanicolaou smear appears to be increased with increasing grade of dysplasia. Interestingly, there does not appear to be an increase in the incidence of residual dysplasia when hysterectomy is performed for positive margin of resection after conization for high grade dysplasia. PMID- 10838300 TI - Poor correlation of high-grade cervical dysplasia with referral cytology: clinical implications. AB - Objective: We questioned the diagnostic utility of cervical cytology by studying whether women with colposcopically guided biopsy proven high-grade cervical dysplasia or cancer had referral cytology results that correlated highly with their disease.Methods: All women with any cytologic abnormality or visible lower tract lesion(s) are referred for colposcopic examination. Ten thousand visits to our colposcopy clinic were prospectively collected, tracked, and reviewed, which revealed 566 cases of high-grade dysplasia and 8 cancers on biopsy. The proportion of high grade disease that was discovered by high grade cytology was measured.Results: The diagnosis of high grade cervical disease was not preceded by high-grade cytology (HGSIL) or "rule out cancer" cytology result (n = 95, 16.8%) in the majority of cases (P <.0001, chi(2)). High-grade disease was more often preceded by low-grade cytology (LGSIL) (n = 224, 39.5%), followed by atypical Papanicolaou smears (ASCUS) (n = 220, 38.9%) and visible cervical or vaginal lesion(s) (n = 27, 4.8%). A very high percentage of cancer cases had preceding HGSIL cytology (6 of 8 cases at least HGSIL, 75%).Conclusion: High grade cervical neoplasia was more likely to be discovered following colposcopic triage of "minor" Papanicolaou smear abnormalities (ASCUS or LGSIL). The false negative rate of a single Papanicolaou smear to detect dysplasia may be as high as 65% (with specificity at the 90% range) (Fahey MT, Irwig L, Macaskill P. Meta analysis of Pap test accuracy. Am J Epidemiol 1995;141:7:680-9), making cytology an inaccurate diagnostic replacement for colposcopy. We should rethink the significance of minimally abnormal Papanicolaou smears when designing triage protocols that delay or eliminate colposcopy based on the Bethesda Grading System alone. PMID- 10838301 TI - Evaluation of the cone biopsy excisor compared with the large loop for electrosurgical excision of cervical lesions. AB - Data incorporated from August 1994 to July 30, 1997.Objective: To compare a newly designed triangular electrode, the Cone Biopsy Excisor to the loop electrode with respect to the margin evaluation, fragmentation, and thermal damage of the conization specimen.Methods: After approval by the Institutional Review Boards of Hartford Hospital, St. Francis Hospital, and New Britain General Hospital/University of Connecticut, patients were randomly assigned to undergo conization with the Cone Biopsy Excisor or with the large loop. Inclusion criteria included biopsy proven CIN II, CIN III, inadequate colposcopy, positive endocervical curettage, or cytohistological discrepancy. Exclusion criteria included pregnancy, undiagnosed uterine bleeding, and invasive carcinoma. The procedures were performed by senior residents on clinic patients at the respective institutions. In addition, after FDA approval November 18, 1996, private OB/GYN practitioners performed cases and added those results to the study. Seventy-eight patients were randomized to the Cone Biopsy Excisor, while 77 patients were randomized to the loop electrosurgical excision procedure (LEEP). To obtain the surgical specimen, Force 2 Valley Lab generators were used with wattage ranging from 35 to 50 according to the size of the instrument chosen, 40/60 blend of coagulation, and cutting current.Pathology reports were reviewed to determine the amount of fragmentation that occurred during the procedure and for tissue diagnosis. Specimens were then evaluated by two gynecologic pathologists blinded to the instrument used. The specimens were analyzed for the ability to evaluate the margins and for degree of thermal damage. A thermal damage score was assigned to each specimen. The scoring system results in 4 levels of thermal damage, from one (minimal thermal damage) to four (heavy thermal damage). Follow-up data was obtained from the patients' medical and pathology records.Results: Seventy-two of 78 (92%) Cone Biopsy Excisor cases vs 10 of 77 (13%) LEEP cases submitted one specimen to pathology, P <.001 based on a chi(2) test. Mean number of specimens submitted to pathology per case in the Cone Biopsy Excisor group was 1.1 +/- 0.5 vs 2.2 +/- 0.8 in the LEEP group, P <.001 based on a t test. Margins were unable to be interpreted because of thermal damage in 2 of 78 (3%) in the Cone Biopsy Excisor group vs 14 of 77 (18%) in the LEEP group, P <.003 based on a chi(2) test.Conclusion: The Cone Biopsy Excisor provided a cervical specimen that had less fragmentation and less thermal damage with margins that were less likely to be indeterminate than those obtained with the large loop electrosurgical procedure. PMID- 10838302 TI - Potential hazards of following atypical and low-grade cervical cytology without colposcopy. AB - Objective: To determine the frequency of high-grade cervical intraepithelial neoplasia in patients with atypical and low-grade cervical cytology and to assess the optimal evaluation and follow-up.Methods: Prospective observational study of 367 of 7,651 private patients who had cytologic, virologic, or colposcopic changes suggesting cervical intraepithelial neoplasia or cervical carcinoma. The study was performed to determine the frequency of cervical intraepithelial neoplasia in the various cytologic groups and to assess the effect of testing for human papillomavirus on the sensitivity, specificity, and positive predictive value of these tests.Results: Papanicolaou smears that included all non-negative tests (high- and low-grade squamous intraepithelial lesions and atypical squamous or glandular epithelial cells of undetermined significance) had the maximal sensitivity (89%) for high-grade cervical intraepithelial neoplasia and cancer. The combined cytologic categories of high- and low-grade squamous intraepithelial lesions had a sensitivity of 58%, this was reduced to 24% if only high-grade squamous intraepithelial lesions were considered relevant for additional evaluation. If we had not evaluated the patients with atypical squamous and glandular cells of undetermined significance, we would have missed diagnosing one third of high-grade and one half of low-grade cervical intraepithelial neoplasia. Cervical cytology was false negative in 8% of patients with high-grade and in 14% of those with low-grade cervical intraepithelial neoplasias. High-risk human papillomavirus deoxyribonucleic acid was detected in 40% of women with high-grade and in 24% of those with low-grade grade cervical intraepithelial neoplasias. The positive predictive value of cytology with atypical squamous cells of undetermined significance increased from 5% to 38% for high-grade and from 30 to 85% for high- and low-grade cervical intraepithelial neoplasias in patients with detectable high-risk human papillomavirus deoxyribonucleic acid. Virologic studies produced no significant improvement on these diagnoses in women with high or low-grade cytology.Conclusions: Because of the poor sensitivity of cytology suggesting high-grade squamous intraepithelial lesions, we recommend that all women with atypical or low-grade cytology be recalled for colposcopy and high risk human papillomavirus deoxyribonucleic acid testing, if available. Decisions whether to perform a biopsy should be based on the result of colposcopic examination. Performing colposcopy only on those patients who have cytologic high grade squamous intraepithelial lesions and following those with lower grade cytologic anomalies without colposcopic diagnosis appears inadequate to rule out high-grade cervical intraepithelial neoplasia. PMID- 10838303 TI - Neural network-assisted ("NNA") analysis of cervical smears: pooled effectiveness results and economic analysis. AB - Objective: To determine the sensitivity of cervical cancer smear screening with neural network-assisted ("NNA") rescreening.Methods: The Papnet system of NNA analysis of cervical smears has been in clinical use worldwide for over 3 years and has been the subject of over 22 published manuscripts reporting on data from over 202,000 smears. This investigation reviewed the results of these studies and classified each study according to study design using a systematic protocol based on reference validation, diagnostic threshold for abnormal, and outcome metric. This classification taxonomy allowed for weighted (based on number of cases in each study) pooling of studies for each study design class. The pooled effectiveness metrics were used to derive the sensitivity of cervical cancer screening with NNA rescreening, using a baseline unassisted screening sensitivity of 85%. Other effectiveness metrics determined by this analysis include NNA's sensitivity as a primary screener, comparisons with primary unassisted screening, and comparisons of NNA rescreening and unassisted rescreening.Results: Analyses of the weighted, pooled mean estimates for each of the principal outcome metrics indicate the sensitivity of cervical cancer screening with NNA ranges from 90% to 99%; most pooled estimates fall in the range of 97-99%. An economic analysis using the APL-based "Cervical Cancer Screen" computer model developed by Eddy (Eddy DM. Screening for cervical cancer. Ann Intern Med 1990;113:214-26) and these effectiveness estimates as inputs showed that NNA analysis involves an accepted level of resource expenditure (approximately $40,000 per life year saved) when added to unassisted screening on a triennial basis.Conclusion: The sensitivity of cervical cancer screening with NNA rescreening using the Papnet system yields sensitivities in excess of 90% and approaching 99%. PMID- 10838304 TI - Comparison of conventional Papanicolaou smears and a fluid-based, thin-layer system for the detection of abnormal cervical lesions: systematic review and meta analysis. AB - Objective: The primary objective of this project was to determine the diagnostic accuracy of a fluid-based, thin-layer system (ThinPrep) compared to conventional Papanicolaou smears for the detection of abnormal (cancerous and pre-cancerous) cervical lesions.Methods: Five comparative studies including 12 study sites, screening centers, and high-risk hospital centers and 10,377 patients were included in a systematic review and meta-analysis examining the cytological diagnoses of ThinPrep with conventional smears using odds ratios (ORs) and risk differences (RDs). All abnormal samples (squamous intraepithelial neoplasia [SIL] or more severe abnormalities) classified by an independent pathologist reviewer were analyzed. Sensitivity analyses using jackknife techniques examined the robustness of the results. Logistic regression models were fit to the data using study level information and the individual patient data to examine the effect of baseline covariates on the results.Results: ThinPrep results were significantly better than conventional smears. For all abnormal diagnoses, the OR = 1.95 (95% CI: 1.40-2.70, P <.0001). For each SIL diagnostic subgroup, the results also strongly favored ThinPrep: LGSIL (OR = 2.20, 95% CI: 1.50-3.22, P =.001) and HGSIL (OR = 1.33, 95% CI: 0.85-2.07, P =.091). Analyses with RDs were similar and also statistically significant. Sensitivity analyses showed the studies to be homogeneous (using Cochran's Q statistic = NS). Logistic regression models examining the meta-analysis results controlling for individual patient level and study level covariates increased the significance of the ThinPrep results.Conclusion: In a meta-analysis of the results of five comparative studies, the ThinPrep system showed significantly higher diagnostic accuracy for the detection of abnormal cervical lesions overall and by SIL diagnostic subgroups. PMID- 10838305 TI - ThinPrep cytology in clinical practice. AB - Objective: To compare ThinPrep cytology to conventional cytology in a gynecologic oncology practice in a university hospital laboratory setting.Methods: From March to September 1997, all Papanicolaou smears in the Division of Gynecologic Oncology were collected with a cervical broom device. A conventional cytology slide was prepared. The broom was then immersed in a Preserv-Cyt and processed. As required for ThinPrep cytology. Managed care patients are not included in this analysis. All cytology specimens were evaluated by the same physician (PE). Increased detection of abnormalities was calculated by the formula increased detection=ThinPrep+-conventional+x100%conventional+Results: A total of 172 split samples were available for analysis. Twenty-two patients (12.7%) had LGSIL or worse. Forty-three patients (25%) had ASCUS or worse cytology.Conclusion: ThinPrep cytology demonstrated marginally improved detection of LGSIL or worse. The extremely high rate of cytologic abnormality and high quality cytologic service in our institution may explain the minimal improvement seen. PMID- 10838306 TI - A large-scale investigator-sponsored field study of the test performance of the ThinPrep papanicolaou test in a low-risk routine outpatient setting. AB - The performance of the ThinPrep Papanicolaou test (TP)(CYTYC Corp., Boxborough, MA) was evaluated in a low- risk outpatient population in order to assess its efficacy in routine clinical use. A total of 18,246 cervical cytology samples were prepared using TP during a 6-month evaluation period from January through June 1997. Results of TP interpretation, assessed for specimen quality and diagnostic information as reported according to the Bethesda system terminology, were reviewed and compared with corresponding results from 2 historical control series of 35,347 and 35,409 conventional cervical smears (Papanicolaou smears). Archived conventional smears obtained during the first 6 months of 1995 and 1996 from the same general patient population by the same group of gynecologists and general practitioners were selected for analysis. Positive cytologic diagnoses in the TP series were reviewed and cytologic-histologic correlations were analyzed when biopsy material was available. The results demonstrated that the proportion of cytologic examinations classified as "satisfactory" was significantly greater in the TP series as compared to conventional controls (88.8% vs 71.1%, P <.0001). There was a significant increase in the number of TP examinations with an adequate endocervical component as compared to conventional controls (90.2% vs 86.9%, P <.005) and a significant decrease in the number of TP examinations that were classified as "satisfactory but limited by" either poor fixation or obscuring factors, such as blood, neutrophils, and contaminants (1.1% TP vs 14.1% nonTP, P <.0001). The overall sensitivity of detection for abnormal cells as indicated by the number of positive diagnoses was increased in the TP series (5.7% vs 2.4%, P <.01). The cytologic diagnoses of ASCUS, LSIL, and HSIL were respectively 1.6, 3.6, and 3 times more frequent in the TP series than in controls. Cytologic-histologic correlations were available for 110 (27%) of the 403 cytologic diagnoses of LSIL and for 90 (83%) of the 121 cytologic diagnoses of HSIL. The presence of a histologic lesion (LSIL or HSIL) was confirmed in 71 (65%) biopsies for LSIL and in 81 (90%) biopsies for HSIL. In conclusion, both sample quality and diagnostic sensitivity were significantly improved using the ThinPrep Papanicolaou test under routine conditions in a low-risk population. PMID- 10838307 TI - Diagnosis of invasive cervical carcinoma with fluid-based, thin-layer slide preparation method. AB - Objectives: Although the main role of the Papanicolaou smear is to detect precursors of cervical cancer, the detection of invasive disease is important to identify women at the earliest stage of their disease. This study assesses the ability of the ThinPrep Papanicolaou test to identify invasive cervical cancer as compared to the Papanicolaou smear.Methods: A meta-analysis of five studies previously reviewed by the Food and Drug Administration was performed. There were 61 cases of cervical carcinoma processed by both conventional Papanicolaou smear and ThinPrep methods. Each case was reviewed by cytotechnologists and cytopathologists without knowledge of prior diagnoses. In a sub-study, the filtrate resulting from the preparation of the ThinPrep was analyzed to determine if diagnostic cells passed through the filter membrane.Results: With the ThinPrep method, a diagnosis of cancer was made in 54 cases (88.5%) as compared to a diagnosis of cancer in 52 of these cases with the conventional Papanicolaou smear method (85.2%). In those cases where diagnosis of invasive cancer was not made with the ThinPrep method, all cases were classified as abnormal, ranging from atypical glandular cells of undetermined significance to high-grade squamous intraepithelial lesions. The ThinPrep slides contained tumor diathesis and other contextual clues. Analysis of the ThinPrep filtrate demonstrated that cancer cells did not pass through the filter.Conclusions: These studies indicate that the ThinPrep method is at least equivalent to the conventional Papanicolaou smear method for the demonstration of squamous cell carcinoma, adenocarcinoma, and other cervical malignancies. Diagnostically important cells are not lost through the filter membrane during the preparation of the ThinPrep slide, because they are larger than the 8 um pore size of the filter. PMID- 10838308 TI - Postmenopausal hormone replacement and obesity. AB - Objective: To compare the association of postmenopausal hormone replacement therapy and lipid and lipoprotein levels in obese and nonobese women.Methods: We studied 4,851 postmenopausal women participating in a population-based observational study. Using cross-sectional data, women were classified into four groups according to their hormone use: current users of estrogen only, current users of estrogen and progestin, never users of hormone replacement therapy, and former users of hormone replacement therapy. Body mass index was categorized as: normal (BMI < 27.3 kg/m(2)), overweight (27.3 kg/m(2) 32.3 kg/m(2)).Results: Hormone use was associated with lower total cholesterol and low-density lipoprotein levels and higher triglyceride levels among all categories of body mass index. However, there was an interaction between hormone use and body mass index for the outcome variable high-density lipoprotein. Among users of estrogen and progestin, high-density lipoprotein levels decreased as body mass index increased. Obese users of combination therapy had a mean high-density lipoprotein level similar to that in obese never users and obese former users of hormone replacement therapy.Conclusion: Our results suggest that, in general, hormone replacement therapy is associated with a favorable lipid profile regardless of body mass index. However, among obese women on combination estrogen and progestin therapy, hormone use was not associated with higher high-density lipoprotein concentrations compared to nonusers. This observation needs further study. PMID- 10838309 TI - Hormone replacement therapy and the risk of colorectal cancer: a meta-analysis. AB - Objective: To review systematically the association between hormone replacement therapy and colorectal cancer incidence and mortality.Methods: We conducted a literature search of English-language studies using MEDLINE, Current Contents, CancerLit, and bibliographies of selected studies. The exposures of interest were ever, recent, or former use of hormone replacement therapy (HRT) and the main outcome measures were colon and rectal cancer incidence and mortality. To reduce the risk of a "healthy estrogen user" bias, we defined recent HRT usage as use either at time of assessment or within the previous year. Studies were evaluated independently by two of the authors. We included studies with adequate controls and retrievable risk estimates. We excluded letters, abstracts, and multiple publications of the same data. The most adjusted risk estimates were extracted. We used the empirical Bayes method, a random-effects model, to calculate summary relative risks (RR) and confidence intervals (CI).Results: Recent use of HRT was associated with a significantly decreased risk of colon cancer (RR = 0.69, 95% CI 0.58-0.81). Protection was limited to recent users; the risk of colon cancer with any use of HRT was 0.94 (95% CI 0.79-1.12). Duration of use was not significant. Three studies addressed colon cancer mortality; the summary RR for death from colon cancer in HRT users was 0.72 (95% CI 0.64-0.81). Rectal cancer incidence and mortality were not associated with HRT use.Conclusion: The risk of colon cancer may be decreased among recent postmenopausal HRT users. Although data are limited, the risk of death from colon cancer may also be lower in HRT users. PMID- 10838310 TI - Clinically favorable effects of raloxifene on bone mineral density and serum lipids: population assessment via bivariate analysis. AB - Introduction: Raloxifene is a nonsteroidal benzothiophene derivative that is classified as a selective estrogen receptor modulator (SERM). It has clinical promise because of its actions as an estrogen-agonist on bone and serum lipids and an estrogen-antagonist on the breast and uterus.Objectives: To determine the percentage of women who respond to raloxifene therapy with regard to bone mineral density (BMD) and serum low density lipoprotein cholesterol (LDL-C) concentration in a large population of healthy postmenopausal women.Methods: Pooled 24-month interim data from two multicenter, placebo-controlled, double-blind phase III studies were used in this analysis. The study population included 1145 healthy women, 45-60 years of age and 2-8 years postmenopausal, randomized to receive either placebo, raloxifene (RLX) 30 mg/day, RLX 60 mg/day, or RLX 150 mg/day. Changes in BMD, measured by dual x-ray absorptiometry, and serum LDL-C concentration, measured by enzymatic assay, were analyzed simultaneously by plotting the median percentage change (baseline to endpoint) in BMD of the lumbar spine and total hip on the y-axis against the corresponding median percentage change in serum LDL-C on the x-axis for each subject in the placebo group and each of the raloxifene groups. The data localized to one of four quadrants on the plot, the upper-left quadrant representing clinically favorable shifts (increased BMD and decreased LDL-C) and the lower-right quadrant representing clinically unfavorable shifts (decreased BMD and increased LDL-C). The bivariate mean, 50th and 95th percentile ellipses, and the percentage of patients improving in one, both, or neither of the measured parameters are displayed.Results: At the end of 24 months, the bivariate mean and the 50th and 95th percentile ellipses of the placebo group for lumbar spine BMD and LDL-C were shifted to the clinically unfavorable quadrant of the plot, reflecting a decrease in BMD and an increase in LDL-C. In contrast, the bivariate mean for all raloxifene groups had shifted to the clinically favorable quadrant of the plot, reflecting an increase in BMD and a decrease in LDL-C. There was a statistically significant difference (Hotelling Lawley test, P <.001) in the bivariate mean for all doses of raloxifene compared to placebo. Only 19.1% of subjects in the placebo group exhibited improvement in both lumbar spine BMD and LDL-C, whereas 43.5%, 47.4%, and 49.8% of subjects in the RLX 30, RLX 60, and RLX 150 group, respectively, exhibited improvement in both of these variables. Furthermore, 35.9% of subjects in the placebo group exhibited no improvement in either lumbar spine BMD or LDL-C, whereas only 13.7%, 10.1%, and 11.6% of subjects in the RLX 30, RLX 60, and RLX 150 groups, respectively, exhibited no improvement in either of these variables. Similar results were obtained comparing total hip BMD and LDL-C.Conclusions: Compared with placebo, raloxifene has beneficial effects on BMD and LDL-C in a large sample of healthy postmenopausal women. Thus, insofar as these intermediate endpoints predict disease outcomes, long-term therapy with raloxifene may be beneficial in the prevention of osteoporosis and cardiovascular diseases. PMID- 10838311 TI - Raloxifene effects on vasomotor and other climacteric symptoms in postmenopausal women. AB - Introduction and Objectives: Raloxifene, a novel selective estrogen receptor modulator (SERM), is under investigation for the prevention of osteoporosis in postmenopausal women. Like traditional estrogen replacement therapy, raloxifene has beneficial effects on bone and on serum lipids whereas, in contrast to estrogen's adverse effects in the breast and uterus, raloxifene is an estrogen antagonist in the breast and is nonstimulatory in the uterus. This study examines the effects of raloxifene 60 mg/day compared with placebo on: 1) the incidence of vasomotor symptoms: hot flashes (flushing) and sweating (including night sweats), 2) the severity and time course of hot flashes, and 3) the relation of hot flashes to baseline subject characteristics and study discontinuations. Additionally, the study explores the effects of raloxifene 60 mg/day compared with placebo on other climacteric symptoms that affect the quality of life of postmenopausal women, such as depression, insomnia, mood lability and genitourinary complaints.Methods: Integrated data from five randomized, placebo controlled studies involving 1,165 healthy, postmenopausal women, with up to 30 months of study drug exposure, were analyzed. The incidence and severity of hot flashes and other climacteric symptoms were compared in patients treated with placebo or raloxifene (60 mg/day) via open-ended, non-directed subject self assessment questionnaires. Data were analyzed for subgroup-by-therapy interactions using many baseline subject characteristics such as age, body mass index, smoking, alcohol, and years post-menopause, as well as preexisting conditions such as hot flashes, sweating, insomnia, depression, and history of hysterectomy. The overall incidence of other climacteric symptoms were reported as adverse events.Results: The increase in overall incidence of hot flashes in raloxifene-treated (24.6%) and placebo-treated (18.3%) subjects was modest, but statistically significant. However, this difference was significant only during the first 6 months of therapy, raloxifene (20.1%) compared with placebo (14.4%). After 6 months of treatment, there was no statistically significant difference in the incidence of hot flashes between the two treatment groups. The majority of hot flashes in raloxifene-treated subjects were subject-assessed as "mild-to moderate" in severity (89%). The incidence of hot flashes reported as "severe" did not differ significantly in raloxifene- or placebo-treated subjects. Subgroup analyses revealed the overall incidence of hot flashes to be highest for both raloxifene and placebo-treated subjects, in younger (age < 55 years) women (P =.004), in women who had previously experienced hot flashes (P =.031), and in women having had hysterectomies (P <.001). Within each of these subgroups, there was no statistical difference in the incidence of hot flashes between the raloxifene and placebo groups. Between the two treatment groups, there was no difference in the overall incidence of subject discontinuations from study due to hot flashes. The occurrence of the other common vasomotor symptom, sweating (which includes night sweats), was not statistically different for the raloxifene or placebo-treated subjects.Genitourinary complaints are often symptoms related to vaginal dryness, such as dyspareunia and decreased libido, as well as other symptoms of vaginitis and leukorrhea. No statistically significant differences occurred for raloxifene- or placebo-treated subjects in reports of these genitourinary symptoms. Similarly, for the other common climacteric symptoms; depression, insomnia, and mood lability, no significant differences in incidence between the raloxifene and placebo treatment groups were observed.Conclusions: Raloxifene (60 mg/day) treatment modestly increased the incidence of hot flashes compared with placebo, however, this difference was only statistically significant during the first 6 months of treatment. There were no differences in the severity of hot flashes between treatment groups, and this symptom did not adversely affect subjects' study participation. In both the raloxifene and placebo treatment groups, young postmenopausal women (age < 55), those with baseline hot flashes, and those with histories of hysterectomy were most likely to experience hot flashes. Raloxifene therapy did not affect the occurrence of other climacteric symptoms commonly affecting the quality of life of women after menopause. PMID- 10838312 TI - Raloxifene HCl is not stimulatory in the endometrium as assessed by the blaustein criteria and an estrogenicity scoring system. AB - Introduction: Raloxifene, a selective estrogen receptor modulator (SERM), acts as an estrogen agonist in the bone and on serum lipids and an estrogen antagonist in breast tissue. The effect of raloxifene on the endometrium of postmenopausal women is a key factor in determining its clinical application.Objectives: The objectives are to determine and compare the histologic outcomes of endometrial samples from healthy postmenopausal women receiving either a high dose of raloxifene or hormone replacement therapy (HRT).Design: The current study is a 24 month, multicenter, double-blind, randomized study of 136 healthy postmenopausal women randomized to receive either raloxifene 150 mg/day (RLX) or continuous combined HRT (Premarin(R) 0.625 mg/day and Provera(R) 2.5 mg/day).Materials and Methods: Endometrial samples obtained by Pipelle biopsy at baseline and endpoint were evaluated using Blaustein's criteria (Kurman RJ, editor Blaustein's pathology of the female genital tract. 1994), which is composed of descriptive diagnostic categories and a newly developed estrogenicity scoring system to quantify subtle morphologic changes in the postmenopausal endometrium (Boss et al. Am J Obstet Gynecol, in press). All subjects with adequate baseline and post baseline endometrial samples were included in the analyses. The results from the 12-month interim analyses are reported and the Blaustein's criteria and the estrogenicity scoring system are compared.Results: Overall, 95.2% of the baseline biopsies were normal. At endpoint 30.6% of the subjects in the HRT group with normal baseline biopsies developed proliferative endometrium and 2.8% developed a polyp, while none in the RLX group developed either. Histological evaluation of stromal and glandular features revealed substantially lower estrogenicity scores in the RLX group. As shown in the table, a high degree of agreement was observed between the estrogen effect grades and the Blaustein's descriptive diagnostic categories at endpoint as shown by a Spearman correlation coefficient of 0.75 and a Kappa coefficient of 0.91.Conclusion: Results from the 12-month interim analyses reveal that, in contrast to HRT, raloxifene does not have stimulatory effects on the endometrium. Also, there is high degree of agreement between Blaustein's criteria and the estrogenicity scoring system used to evaluate endometrial histology. PMID- 10838313 TI - Pregnancy rates following hysteroscopic polypectomy, myomectomy, and a normal cavity in infertile patients. AB - Objective: To assess the reproductive benefits of hysteroscopic myomectomy and polypectomy for infertility when compared to infertile couples with a normal cavity at hysteroscopy.Material and Methods: All patients with a diagnosis of infertility who underwent hysteroscopic evaluation by a single surgeon between 1975 and 1996 were sent a questionnaire as routine follow-up regarding their reproductive history. All 100 subjects who were located responded to the questionnaire, and 78 subjects met the inclusion criteria; age <45 years, 12 months of infertility, and 18 months of follow-up with attempts to conceive including in vitro fertilization in patients with bilateral tubal occlusion.Results: Of the 78 subjects, 36 had undergone a myomectomy, 23 a polypectomy, and 19 had a normal cavity. Among the three groups there was no significant difference in their ages, types of infertility, length of infertility, or follow-up after the procedure. Using the Cox proportional hazard model, and adjusting for age, polypectomy patients had a significantly higher pregnancy rate (RR 3.89, P <.01) and a higher live birth rate (RR 2.42, P =.06) than patients with a normal cavity. Patients who had undergone a myomectomy also had a higher pregnancy rate (RR 2.02, P =.11) and live birth rate, but this did not achieve statistical significance. Pregnancy following a hysteroscopic myomectomy was associated with a larger fibroid resection (3.15 cm vs 2.5 cm P =.05). The spontaneous abortion rate following the myomectomy, polypectomy, or a normal study was equivalent, 28.1%, 23.1%, and 29.2%, respectively.Conclusions: Both hysteroscopic polypectomy and hysteroscopic myomectomy appear to enhance fertility when compared to infertile patients with a normal cavity. Despite concern that hysteroscopic resection of a large myoma may ablate a large surface area of the endometrial cavity, patients with larger myomas were more likely to conceive following resection. PMID- 10838314 TI - Tubal surgery vs in vitro fertilization for the treatment of infertility due to distal tubal occlusion. AB - Background: The optimal treatment of infertility due to tubal occlusion has not been established. Many practitioners feel that the success of tubal repair exceeds that of in vitro fertilization (IVF); however, previous studies of pregnancy after tubal surgery have been limited by bias in patient selection, follow-up, or surgical expertise. The purpose of the present study was to determine the outcome after repair of distal tubal occlusion performed by experienced surgeons in an unselected patient population with consistent follow up.Design: Chart review with telephone contact of patients lost to follow up.Methods: The records of all tubal surgery performed between 1989 and 1996 at the University of Alabama Hospital and The Kirklin Clinic outpatient surgery facility were reviewed. All women with infertility due to distal tubal occlusion, with or without pelvic adhesions, who had no other significant infertility factors were included for study. Details of the infertility history, operative procedure, and postoperative course were recorded. Patients lost to follow-up within 1 year after surgery were contacted by telephone for information regarding subsequent testing and treatment and pregnancy outcome.Results: Eighty-three women aged 19-39 years met the entry criteria for this study. Follow-up of at least 1 year was obtained in all but 11 patients. Tubal surgery was accomplished by laparotomy in 19 women; 64 women underwent tubal repair by laparoscopy. Within 1 year of surgery, 9 hysterosalpingograms, 51 clomiphene cycles, and 20 gonadotropin cycles were performed on the study group. Pregnancy was achieved within 1 year in 13 women; of these, there were 6 live births (9.6% birth rate per surgery), 2 spontaneous abortions, and 3 ectopic pregnancies. There were no live births among women who underwent tubal repair by laparotomy. None of the postoperative gonadotropin cycles resulted in pregnancy. Seven women underwent IVF within 1 year after surgery because of extensive tubal damage noted at surgery. Based on current charges for the infertility treatments performed, the cost of a live birth with tubal surgery exceeded $120,000, versus less than $50,000 per live birth with IVF using results obtained nationally or at UAB.Conclusions: The cost-effectiveness of reconstructive surgery in unselected patients with distal tubal occlusion is less than that of IVF. Empiric use of gonadotropins for ovarian stimulation does not improve pregnancy rates after tubal surgery. In our series, laparoscopic tubal repair seemed to give results superior to that of laparotomy. PMID- 10838315 TI - Outcome of in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) using human sperm prepared via a new standardized swim-up technique fit for office use. AB - Objective: Recent developments in the assisted reproductive technology (ART) areas necessitate the use of new and more efficient and acceptable modes of gamete in vitro manipulation techniques. As established via the use of various forms of ART, the natural process of fertilization has been largely bypassed and the gametes are obtained and manipulated outside of the normal means of conception. For the male, improvements in sperm quality have involved the use of various techniques that aim to recover a small percentage of healthy spermatozoa that can be further used for the different ART procedures. Previous IVF results showed no differences in fertilization rates between ZSC and Percoll recovered sperm (Zarmakoupis-Zavos et al. ACOG, 1997). The current study was designed to study the fertilizing capacity of ZSC recovered spermatozoa and subsequent embryonic development and pregnancy rates established via IVF and ICSI procedures.Methods: Sperm specimens (n = 30) were prepared via the use of the standardized ZSC swim-up technique, which was designed to harvest almost all overlayered media at the end of the procedure to maximize the sperm recovery. The ZSC device consists of a column with a conical cavity where the semen is placed, and from where the sperm swim-up into the overlayered media and are subsequently recovered at the end of the procedure. Semen specimens were initially assessed and then prepared (0.5 mL each) via the ZSC swim-up technique. The enhancement of the routine semen parameters was assessed and the recovered specimens were used for either conventional IVF or ICSI.Results: The results of the fertilization and pregnancy via IVF and ICSI are shown below:The generated results point out that ZSC recovered specimens yielded acceptable levels of fertilization and pregnancy rates in our IVF and ICSI program. Furthermore, the IVF inseminations yielded higher fertilization rates with a slight increase in polyspermic oocytes when compared to the ICSI inseminations.Conclusion: The data generated in this study tend to point out that the ZSC, besides being a simple one-step standardized system, can also be effectively employed in an ART program and can generate adequate fertilization and pregnancy rates. We have previously shown that the quality and fertilizing capacity of ZSC recovered sperm is comparable to that recovered via Percoll gradients. However, the ZSC when compared to the Percoll method was less time consuming (5 minutes of actual labor) and less tedious by eliminating the dilution and centrifugation steps of Percoll preparation, since it washes the sperm while simultaneously selecting it. The ZSC is a standardized semen preparation method and could be the method of choice in an ART program. PMID- 10838316 TI - The effects of in vitro fertilization on subsequent pregnancy outcome. AB - Objective: The purpose of this study was to look at the effects of assisted reproductive technology on subsequent pregnancy outcome variables.Methods: This retrospective study was performed using the Regional Perinatal Data System, a population-based birth registry. The data system is used by all 23 hospitals in the 14-county region of Central New York. 42,656 births were analyzed from January 1, 1995 through December 31, 1996. The associations between in vitro fertilization and premature rupture of membranes (PROM), preterm delivery (PTD), age, gravidity, parity, and race were evaluated using chi(2) and Fisher's two tailed Exact analyses. Odds ratios represent the magnitude of the association.Results:Women undergoing in vitro fertilization were significantly more likely to experience PTD and PROM. They were significantly more likely to be nulliparous, primigravidas, or over the age of 34.Conclusions: This analysis shows a strong association between women who have undergone in vitro fertilization and adverse pregnancy outcomes including PROM and preterm delivery. This study suggests a need for intensified obstetric care and the assignment of high risk pregnancy status to those women who have undergone in vitro fertilization. This association has not been previously reported and underscores the need for a comprehensive data system to analyze adverse events within a region and develop strategies for prevention. PMID- 10838317 TI - Human motile sperm recovery after cryopreservation: freezing in nitrogen vapor vs the direct plunge technique. AB - Objectives: The following will be tested: 1) Freezing in nitrogen vapor prior to submerging will have a higher motile sperm recovery rate compared to directly submerging in liquid nitrogen, 2) The length of time a sample spends in nitrogen vapor will not affect the motile sperm recovery, and 3) The volume of sample to be frozen will not affect the recovery of motile sperm.Design: This is a prospective study performed through a university infertility clinic. The semen samples were obtained from 20 male partners of couples being screened for in vitro fertilization.Materials and Methods: Semen was collected and washed with 5 volumes of modified Ham's F10 medium and centrifuged at 120xg for 10 minutes. The pellet was resuspended in 5 mL of medium and centrifuged in a similar manner. The swim-up sperm was prepared with the second pellet overlaid with 1 mL of medium, containing 1% human serum albumin, and incubated for 1 hour at 37 C. Equal volumes of sperm suspension and cryoprotectant were mixed by gentle stirring. Sperm density and motility were recorded before freezing. Each vial contained 1 mL of the sperm/cryoprotectant mixture. All vials were eventually submerged in liquid nitrogen for 40 minutes and thawed at room temperature for 1 hour. The percent of motile sperm recovered was assessed. Hypothesis 1: Comparing freezing in nitrogen vapor versus direct plunge, one vial was frozen for 6 minutes in the nitrogen vapor 2 cm above the liquid surface prior to submerging. The paired vial was kept at room temperature for 6 minutes before submerging. Hypothesis 2: Two vials were frozen in vapor for 6 and 12 minutes prior to submerging them. Hypothesis 3: To assess the effect of sample volume on motile sperm recovery, three separate vials were prepared, each containing either 1.0, 0.5, or 0.25 mL of the sperm/cryoprotectant mixture. Each vial was suspended in vapor for 6 minutes prior to submerging into the liquid. Paired t test analysis was used for objectives 1 and 2, with repeated measures analysis of variance used to determine the significance of objective 3.Results: Pure motile sperm density and motility were assessed for 20 samples after the addition of cryoprotectant. The sperm density and motility ranged from 0.9 to 120 million sperm/mL (33.2, SD +/- 35.0) and 78-100% (86.5, SD +/- 12.4), respectively. The mean motile sperm recovered for vapor freezing prior to plunge was 54.9% (SD +/- 15.4), compared to submerging directly, which was 21.5% (SD +/- 10.0); this was significant with a P <.05. The initial sperm density, <20 or >/=20 million/mL, did not influence the percent of motile sperm recovered (P >.05). Motile sperm recovered from samples in vapor for 6 and 12 minutes was 49.7% (SD +/- 8.71) and 51.8% (SD +/- 10.4), respectively; this was not significant. The volume of purified motile sperm frozen did not significantly alter the percent of motile sperm recovered.Conclusion: Directly submerging a sample from room temperature into liquid nitrogen has shown a reduction of motile sperm recovery by 65.0%, compared to exposing the sample to liquid nitrogen vapor prior to submerging, which only showed a reduction of 31.7%. Motility appears to be influenced by the rate at which the sample is frozen. There was no significant impact on motile sperm recovery compared to the amount of sperm preserved, length of time the sample was in liquid nitrogen vapor, or the initial sperm density. This is the first study to isolate the motile faction from semen and examine the recovery rate between the two cryopreservation techniques. The clinical application implies that increasing the recovery of motile sperm could improve pregnancy rates and this simplified technique is possible without obtaining special equipment that could impose a significant financial burden with similar results. PMID- 10838318 TI - Viability of human semen specimens cryostored and transported at 5 C using the Bio-Tranz shipping system. AB - Objective: Transport of unprocessed human semen specimens from the production site to distant laboratories for andrological evaluation and clinical use requires the development of proper protocols and devices for the shipment and maintenance of sperm viability during transport. Factors such as maintenance of proper temperature and the specific diluent used are considered to affect the viability of semen specimens during transport. The Bio-Tranz shipper (ZDL, Inc., Lexington, KY, USA), which was designed to cool specimens (5 C) during transport, consists of a properly refrigerated Styrofoam box, TEST-yolk Buffer (TYB), a nonspermicidal condom-shaped semen collection kit (Hygene Kit; ZDL, Inc.) and test tubes (15.0 mL). The viability of semen specimens stored from the time of collection to the time at which the specimens were to be processed and used (24 hours post-collection) was evaluated using the Bio-Tranz shipping technology.Methods: Semen specimens (n = 30) were assessed for percentage and grade of motility (0-4), and for the sperm membrane functional integrity as measured by the hypoosmotic swelling (HOS) test at collection time and 24 hours after storage in the Bio-Tranz shipper. The specimens were produced at intercourse via the use of the Hygene collection kit, assessed for sperm characteristics, split into two aliquots and then transferred to 15.0 mL test tubes. Aliquot 1 was mixed 1:1 (v/v) with TYB media and aliquot 2 was used as raw (unprocessed). The specimens were then placed and secured in the Bio-Tranz shipper and assessed for sperm qualitative characteristics following 24 hours of storage.Results: The results of the sperm parameters assessed among the various seminal treatments are shown below:Sperm characteristics were significantly improved when preparing the specimens using TYB (0 h; P <.05). Significant differences in all sperm qualitative characteristics assessed were noticed between raw specimens and those prepared via TYB after 24 hours of storage (P <.05). Most interestingly, sperm characteristics between raw specimens (0 h) and specimens prepared using TYB and stored via the use of the Bio-Tranz shipper for 24 hours were not different (P <.05).Conclusion: The results obtained show that collection and shipment of semen specimens via the Bio-Tranz shipper system is possible. The Bio-Tranz shipper maintains adequate sperm viability after 24 hours of cryostorage. The use of the Bio-Tranz shipper is extremely convenient for patients that request semen processing services such as semen cryostorage, semen evaluation, semen preparation for IUI purposes, or other assisted reproductive technologies. The technique could be of significant clinical and economic importance to the patient and to the treating physician at locations across the United States. PMID- 10838319 TI - Depo-provera: an excellent contraceptive for those who continue to use it. AB - Objective: This paper focuses on the user profile, side effects, and discontinuation rates of Depo-Provera users at Grady Memorial Hospital, a large inner-city hospital in Atlanta, Georgia.Methods: Between July 1993 and April 1996 baseline and follow-up interviews were conducted with African-American and Caucasian women who were using a contraceptive method. Women had to choose a method that they had not used in the previous 3 months.Results: Depo-Provera was one of the top two contraceptive methods chosen at Grady Memorial Hospital. Convenience and effectiveness were the main reasons for its selection. Of the total Grady Hospital sample (n = 1,346), 404 women (30%) selected Depo-Provera as their method of contraception. Approximately 70% of the Depo-Provera users were aged 16-25 years, African American (98.3%), had never been married (88%), were on Medicaid (73.5%), and had had at least one pregnancy (94.3%). Depo-Provera users experienced menstrual (92.6%) and non-menstrual (67.6%) side effects. Menstrual side effects included amenorrhea, irregular cycles, spotting, and long menses. The most prevalent non-menstrual side effects were weight gain and headaches.The 12-month discontinuation rate of Depo-Provera was 49.2%, compared to oral contraceptives (66%) and Norplant (15%). The main reason cited for discontinuation of Depo-Provera was non-menstrual side effects (35.6%), menstrual side effects (23.1%), and inconvenience (12.0%). Of all women who initiated Depo Provera use, 11.0% were pregnant at 12 months and 16.5% became pregnant by the first follow-up survey (average of 17 months).Conclusions: At Grady Memorial Hospital, Depo-Provera was a popular birth control method with high discontinuation rates. Menstrual, non-menstrual side effects, and inconvenience were the chief discontinuation factors. The impact of Depo-Provera discontinuation upon the pregnancy rate is substantial. PMID- 10838320 TI - Experience and acceptability of emergency hormonal contraception. AB - Objective: The study was designed to evaluate the experience and acceptability of emergency hormonal contraception service for inner-city women in Planned Parenthood clinics.Methods: In 1995, emergency hormonal contraception was introduced as a new service in three clinics of New York City's Planned Parenthood that served low-income women. Shortly after inception of this service, the agency undertook a survey to investigate patient experience and acceptability of the service. The survey addressed side effects and patients attitudes about emergency hormonal contraception. It also addressed suggestions for improving the service.The indication for the use of this method was unprotected intercourse within the previous 72 hours of first dosage of certain birth control pills. We used Lo/Ovral (norgestrel 0.3 mg + ethinyl estradiol 30 ug) 4 tablets given stat and second dose 12 hours later.Results: There were 118 patients who responded to the survey. The average patient age was 25 years. Approximately half of the women were white and half were other. The majority were nulliparous, and 42% had one or more abortions in the past. Side effects such as nausea, vomiting, and fatigue and their prevalence were noted. Two women reported method failure. Both patients elected abortion. The vast majority (92%) stated that they would use the method again. Patients also made several recommendations for enhancing the service.Conclusion: As a result of this survey, Planned Parenthood of New York City completely revised its emergency hormonal contraception protocols in order to increase patient satisfaction while maintaining high medical standards to ensure lower method failure and drug side effects in more than 1,000 patients so far. PMID- 10838321 TI - Differences in contraceptive methods used by women with physical disabilities compared to women without disabilities. AB - Women with physical disabilities face unique challenges often not addressed by gynecologists regarding choices of contraceptive method. Interactions between some hormonal methods of contraception and disability-related medications, inability to use barrier methods because of limitations in manual dexterity, potentially elevated risk for DVT, and need for menstrual management are factors that complicate decisions about contraceptive methods for this growing population.Objective: This study assessed similarities and differences in the use of and satisfaction with different methods of birth control between women with disabilities and women without disabilities, while controlling for age at onset and severity of disability.Methods: As part of a national survey, 616 women of childbearing age (315 with a variety of physical disabilities, 301 without disabilities) reported their use of and satisfaction with various forms of contraception.Results: Women with physical disabilities were significantly less likely than women without disabilities to use hormonal (11.7%/19.6%) or barrier methods (11.1%/17.6). They were significantly more likely to have had a hysterectomy (8.9%/4.3%) or use no method (41.9%/33.4%). There were no significant differences in the use of surgical methods (22.9%/22.6%) or natural methods (3.5%/2.7%). Women with more severe disabilities were significantly more likely to have had hysterectomies or use surgical or no method of contraception. Women with disabilities were least satisfied with barrier methods.Conclusion: Women with disabilities are more likely to use permanent methods or no method of contraception. Further research is needed to understand the impact of functional limitations and other disability-related factors on the use of other contraceptive methods in this population. PMID- 10838322 TI - A multicenter, randomized comparative trial of the clinical effects on cycle control of two 21-day regimens of oral contraceptives containing 20 ug EE. AB - Purpose: This multicenter, randomized, open-label comparative study was designed to evaluate the effects on cycle control of the only two oral contraceptives (OCs) containing 20 ug ethinyl estradiol (EE) that are available in North America.Methods: Women of childbearing age who desired OCs for birth control were eligible to enroll in this study. Sixty-two subjects were randomly assigned to receive Alesse (levonorgestrel 0.1 mg and EE 20 ug) or Loestrin Fe 1/20(R) (norethindrone acetate 1.0 mg and EE 20 ug).Results: At the time of the interim analysis data cut-off 32 subjects had completed 3 cycles of Alesse and 30 had completed 3 cycles of Loestrin Fe 1/20. The groups were similar with regard to age, weight, height, race, and prior OC usage. A total of 22% of the subjects in the levonorgestrel group and 27% of those subjects in the norethindrone group had never used OCs. The remainder had not used OCs for at least 3 months before entering the trial. During the third cycle of use, the percentage of normal cycles was significantly greater in the levonorgestrel group (P <.05). Moreover, the rate of intermenstrual bleeding (breakthrough bleeding and/or spotting) in the norethindrone group (62.9%) was more than double that in the levonoregestrel group (29.0%). Cumulatively, from cycles 1 to 3, there was a higher incidence of absence of withdrawal bleeding with norethindrone when compared with levonorgestrel (27.7% vs 5.3%).Conclusion: The results of this study demonstrate that good cycle control can be achieved with an OC containing 20 ug EE. The superior cycle control of Alesse compared with Loestrin Fe 1/20 is consistent with that found in comparative studies of the cycle control of levonorgestrel, norethindrone, and norethindrone acetate containing OCs that had a higher dose of EE. PMID- 10838323 TI - The tolerability of a triphasic norgestimate/EE-containing OC: results from a double-blind, placebo-controlled trial. AB - Objective: To compare adverse event data collected during administration of a triphasic norgestimate ethinyl estradiol (EE) containing oral contraceptive (OC) [ORTHO TRI-CYCLEN(R)] or placebo.Methods: Four-hundred fifty-two females between the ages of 15 and 49 years were enrolled in two multicenter, double-blind, randomized, placebo-controlled studies. These studies were conducted to evaluate the effectiveness of triphasic norgestimate/EE in the treatment of acne vulgaris (Redmond G, Olson W, Lippman J, et al. Norgestimate and ethinyl estradiol in the treatment of acne vulgaris: a randomized placebo-controlled trial. Obstet Gynecol 1997;89:X-X; Lucky A, Henderson T, Olson W, et al. The effectiveness of norgestimate and ethinyl estradiol in treating acne vulgaris. J Am Acad Dermatol, in press). Participants returned to the study centers monthly for evaluation. At each visit, interviews were conducted to elicit and record the occurrence of adverse events. Reports were derived by nondirected questioning.Results: The table below describes the number and percentage of subjects reporting the event during the 6-month period. Note that, over the range of percentages observed, the differences detectable with a power of 0.80 range from approximately 4% to 12%.Conclusion: Excellent tolerability for the triphasic norgestimate/EE OC is demonstrated by a low rate of side effects that did not differ in a statistically significant manner from placebo. PMID- 10838324 TI - An analysis of risk factors associated with blood transfusion in ectopic pregnancy. AB - Objective: To determine the risk factors associated with blood transfusion in ectopic pregnancy.Methods: A retrospective chart review of the presentation and hospital course of ectopic pregnancies managed over 5 years at two hospitals was undertaken. Thirty-two variables, including demographics, presenting signs and symptoms, and intraoperative findings, were examined in univariate and multivariate logistic modeling.Results: Among 185 patients with histologically confirmed ectopic pregnancies who were managed surgically, 8.6% or 16 women required transfusion. Multivariate analysis of risk factors for blood transfusion demonstrated a statistically significant association with 1) initial hemoglobin <10 g/dL (odds ratio [OR] 38.8, 95% confidence interval [CI] 6.0-356.8) and 2) hCG levels >/= 6500 mIU (OR 18.1, 95% CI 3.6-158.1), as well as 3) abnormal bleeding on presentation (OR 0.08, 95% CI 0.007-0.42. Presence of two of these factors has a sensitivity of 82% (95% CI 48-98%) and a positive predictive value of 33% (95% CI 16-54%). No case had all three factors present.Conclusion: This report is, to our knowledge, the first regression analysis of risk factors for transfusion associated with ectopic pregnancy. It demonstrates that initial hemoglobin and hCG levels as well as abnormal bleeding on presentation are independent risk factors for blood transfusion in ectopic pregnancy. PMID- 10838325 TI - Management of separation pain after single-dose methotrexate therapy for ectopic pregnancy. AB - Objective: To review the success of conservative management of moderate to severe abdominal/pelvic pain occurring after treatment of ectopic pregnancy with systemic methotrexate, to evaluate prognostic factors for success, and to determine if the overall resolution time was shorter in such patients.Methods: A retrospective chart review of all single-dose methotrexate patients treated from January 1, 1992 to January 1, 1997 who were admitted for observation or evaluated and subsequently discharged after an episode of increased abdominal/pelvic pain unrelieved within 1 hour by 800 mg oral ibuprofen. Before 1992, patients developing such pain generally underwent surgery. Candidates for conservative management were hemodynamically stable and had no more than moderate (confined to the pelvis) free fluid. Mild rebound was not an exclusion. Hospitalized patients had serial abdominal examinations, hematocrits, and hCG titers. Hematocrits, ultrasound findings, hCG levels, time for hCG levels to reach /= 2.1 ng/mL) predicted delivery prior to 37 weeks gestation (P <.0003; OR 3.4, 95% CI 1.7-6.8). If a rescreen test at one week was also >/=2.1 ng/mL (8.9% of all subjects) then relative risks of preterm birth increased to 6.86 CI (3.2-14.5) P <.0001. Having two consecutive positive tests was associated with a mean time to delivery of 2.3 weeks (negative predictive value of 97%). 2) Survival analysis of women delivering at term and preterm demonstrated that sE(3) >/= 2.1 ng/mL predicted onset of labor and birth in approximately 3 weeks:Conclusion: Meta-analysis of these two large prospective trials confirms that 1) an sE(3) >/= 2.1 ng/mL provides timely warning of a presumed endocrine signal for impending preterm and term parturition, 2) the 2-3 week mean time to delivery after a positive signal allows time for intervention, and 3) lack of a positive sE(3) signal allows a confident prediction (NPV) that delivery will not occur within 2-3 weeks. PMID- 10838335 TI - Perinatal outcome of triplet gestation: does prophylactic cerclage make a difference? AB - Objective: To compare the perinatal outcome of triplet gestations with and without prophylactic cerclage.Methods: A retrospective chart review of all triplet gestations delivered between January 1988 and June 1997 was performed. Only women initiating prenatal care before 15 weeks gestation were included. Management guidelines for triplets were uniform with the exception of use of prophylactic cerclage placed at 14 +/- 1 weeks gestation. Hospitalization was reserved for maternal medical and obstetric conditions, or fetal indications. Tocolytics were initiated only for persistent uterine contractions with intact membranes. All deliveries were effected by cesarean section at 37 weeks or less for maternal or fetal indications. All neonates were cared for at our institution. The cerclage group was compared to the no cerclage group for gestational age at delivery and maternal and neonatal complications. Descriptive statistics, Student t test, and Mann-Whitney U test were used for analysis.Results: Fourteen of the 51 (27.4%) sets of triplets had cerclage. There were no differences between groups when compared for maternal age, length of hospital stay, preterm labor rate, steroid use, histologic or clinical chorioamnionitis, and postoperative complications. In addition, there were no differences in mean birth weight, Apgar scores, respiratory distress syndrome, intraventricular hemorrhage/periventricular leucomalacia, NICU stay, and neonatal mortality. Although the mean gestational age at delivery for the cerclage group (33.1 +/- 2.7 weeks) was not different from the no cerclage group (31.7 +/- 3.7 weeks), the proportion of pregnancies delivered at 31 weeks or more was significantly higher in the cerclage group (13/14 vs 23/37, P =.03).Conclusions: Prophylactic cerclage may influence the proportion of triplet gestations delivered at 31 weeks or more. Lack of differences in the other parameters may be reflective of the small sample size. PMID- 10838336 TI - Effects of magnesium sulfate tocolysis on maternal mental status. AB - Objective: Magnesium sulfate is the most commonly used agent for tocolysis in the management of preterm labor. Anecdotally both clinicians and patients have noted alterations in mental status of women receiving high doses of magnesium infusion, and central nervous system depression including lethargy and depression of deep tendon reflexes has been documented. Our hypothesis is that intravenous magnesium sulfate at tocolytic doses significantly affects maternal mental status.Materials and Methods: Eligibility criteria included admission or transport to University Hospital between 25 and 34 weeks gestation with an initial episode of spontaneous premature labor. Patients were consented and given an initial mental status examination prior to magnesium sulfate infusion. Those patients transported to University Hospital diagnosed with premature labor and already on magnesium were consented and given an initial mental status examination at the time of arrival. Once a therapeutic level of magnesium was documented a repeat mental status examination was performed, with a third examination performed 24 hours after the magnesium infusion has been discontinued. As a control group pregnant women hospitalized for premature rupture of the membranes not in labor had the examinations performed initially, after 24 hours of hospitalization, and again after 72 hours of hospitalization. Exclusion criteria included underlying mental illness, administration of other medications that might affect mental status, cervical dilation greater than 4 cm, clinical evidence of chorioamnionitis, or the presence of any significant abnormalities in the fetal heart rate tracing. The mental status examination consisted of the mini mental state exam, the comprehension portion of the Wechsler Adult Intelligence Scale, and the Bender Gestalt Indicator. The results of these examinations were scored in a blinded fashion by a psychiatrist.Results: There were 22 patients in the study group and 9 patients in the control group. There were no differences in the age, gravidity, parity, or gestational age of the two groups. Out of a possible 102 points, the mean mental status scores were as follows:The mean serum magnesium level at the time of therapy was 5.1 mg/dL. The time required for response to the Wechsler test was significantly different although the scoring of the comprehension was unchanged (15.2 +/- 3.6 min vs 22.3 +/- 4.7 min, P <.05).Conclusions: This prospective blinded study reveals no differences in maternal mental status during magnesium sulfate infusion at the levels recorded in our study. There was an increase in the length of time required to answer the comprehension and judgment portion of the examination. These findings have significant clinical implications suggesting that patients on magnesium sulfate can make appropriate judgments and can, therefore, participate in clinical discussions and trials. PMID- 10838337 TI - Risk factors in premature rupture of membranes. AB - Objective: This retrospective study was undertaken to investigate risk factors in women who have preterm premature rupture of membranes. This information will aid the clinician in targeting at-risk women for intensified obstetric care and entry into prevention programs.Methods: 28,725 deliveries were analyzed over a 16-month time frame (January 1, 1995-April 30, 1996). These data were collected via a 14 county, 23 hospital population based Perinatal Data System. The associations between premature rupture of membranes and risk factors were analyzed using chi(2) and Fisher's Exact test analyses. Odds ratios show the magnitude of these associations.Results: Risk factors included vaginal bleeding, tobacco usage, multiple births, polyhydramnios, maternal age <20 years and >34 years, congenital malformations, parity, race, in vitro fertilization, sexually transmitted disease, prior preterm delivery, and incompetent cervix. Premature rupture of membranes shows a significant association with prior preterm delivery, prior history of a low birth weight infant, incompetent cervix, tobacco use (12 pack per day), multiple births, parity, in vitro fertilization, sexually transmitted diseases, and viral diseases, P <.001 (for all of these risk factors), polyhydramnios and non-white race, P <.01.Conclusions: These associations help to develop a picture of the patient at risk for premature rupture of membranes. Strategies can be better developed to enter high-risk patients into prevention programs to maximize pregnancy outcomes. PMID- 10838338 TI - Preterm premature rupture of the membranes is associated with asymmetric intrauterine growth retardation and smaller placentas. AB - Objective: To show that infants delivered prematurely because of preterm premature rupture of the membranes (PPROM) show a tendency for asymmetric intrauterine growth retardation (IUGR). At the same time, to demonstrate that these pregnancies exhibit nutritional deprivation by the presence of correspondingly smaller placentas, as demonstrated by the placental-fetal ratio.Study Design: A prospective study was performed over 2 years comparing the tendency for IUGR in infants born of pregnancies with PPROM (N = 86), as compared to normal term controls (N = 351). The tendency for growth restriction in the neonate was determined using the ponderal index at birth. Exclusion criteria included pregnancies complicated by diabetes, hypertension, preeclampsia, multiple gestations, genetic, and other recognized causes of IUGR. Four controls were selected for each study patient who delivered the same day. The mean ponderal index (PI(m)) was compared, as were the mean placental-fetal weight ratios (PWt/PI)(m), for both groups. A comparison of the PI(m) and (PWt/PI)(m) for infants delivered prematurely with and without rupture of membranes was also noted. Analysis was by the paired Student's t test and chi(2) test.Results: The newborn PI was found to be independent of maternal age, gravidy, and parity. The number of male and female infants were not significantly different in the control and study groups. The PI(m) for the PPROM group (PI(m) = 2.33, SD = 0.29) was significantly smaller than that for the controls (PI(m) = 2.52, SD = 0.23) (P <.0005). Similarly the (PWt/PI)(m) for the PPROM group (mean = 194.8, SD = 49.9) was significantly smaller than that for the control group (mean = 273.4, SD = 54.2) (P <.0005). Mean values of PI and PWt/PI for preterm deliveries with, as compared to without rupture of membranes, showed no significant differences.Conclusions: Using the ponderal index to detect IUGR, a significant association of IUGR is noted in infants delivered subsequent to PPROM. This growth impairment is accompanied by significantly smaller placentas. PMID- 10838339 TI - A randomized trial comparing prostaglandin E(2) vaginal insert (Cervidil) to vaginal gel for cervical ripening before induction of labor. AB - Objective: To compare the efficacy and safety of a controlled release vaginal insert (Cervidil) with hospital compounded prostaglandin E(2) vaginal gel for cervical ripening before labor induction.Methods: A prospective randomized study was performed between July 1996 and August 1997 on 200 patients undergoing cervical ripening before induction of labor. The efficacy and safety of a 12-hour controlled release 10-mg vaginal insert (Cervidil, Forest Pharmaceuticals) was compared with two 4-mg doses of prostaglandin E(2) gel.Results: The study groups did not differ by parity, estimated gestational age, indications for induction, and initial Bishop scores. No differences were noted in Bishop score at 12 hours or change in Bishop score. No statistically significant differences were noted in the successful inductions, additional days of induction, and the number of patients discharged undelivered. There were no differences in mean time to active labor, cesarean section, meconium staining, 1 and 5 minute Apgar scores, cord pH values, NICU admissions, and incidence of hyperstimulation.Conclusions: Two 4-mg doses of prostaglandin vaginal gels are equally effective to a Cervidil vaginal insert for preinduction cervical ripening. PMID- 10838340 TI - Comparative cost analysis of prostaglandin analogues dinoprostone and misoprostol as labor preinduction agents. AB - Objective: To compare the relative cost efficacy of three commercially available prostaglandin analogues, misoprostol (Cytotec(R), PGE(1)), dinoprostone gel (Prepidil(R), PGE(2)), and dinoprostone pessary (Cervidil(R), PGE(2)), as labor preinduction agents.Methods: The investigation was conducted as a prospective randomized, blinded phase III clinical trial. One hundred twelve gravid females undergoing induction of labor with an unfavorable cervix (Bishop score /=6) after the initial preinduction interval in the Cytotec(R) (79.0%) and Cervidil(R) groups (60.5%) as compared to Prepidil(R) (40.0%) (P <.05). The average relative cost per patient for prostaglandin preinduction was significantly less with Cytotec(R) ($0.42) as compared to both Cervidil(R) ($168.00) and Prepidil(R) ($184.17) (P <.0001). A greater percentage of patients treated with Cytotec(R) (42.1%) achieved active labor and delivered without the use of oxytocin when compared to Cervidil(R) (15.8%) or Prepidil(R) treatment groups (2.9%) (P <.05). Additionally, time to delivery (+/-SD) was significantly shorter in the Cytotec(R) group (24.0 +/- 10.8 h) as compared to the Cervidil(R) (32.2 +/- 14.7 h) or Prepidil(R) groups (33.9 +/- 16.2 h) (P <.05). This reduction in time resulted in a significantly lower overall mean cost per patient incurred by labor induction in the Cytotec(R) group ($723.66) as compared to the Cervidil(R) ($1058.90) or Prepidil(R) treatment ($1124.84) (P <.01). No significant differences were noted with respect to rate of cesarean delivery.Conclusion: Misoprostol is more cost-effective than the comparable commercial dinoprostone prostaglandin preparations as an adjuvant to labor induction in patients with an unfavorable cervix. PMID- 10838341 TI - Attempted vaginal birth after cesarean section: a multicenter comparison of outpatient prostaglandin E(2) gel with expectant management. AB - Objective: To compare the clinical effectiveness and safety of outpatient administration of an intracervical prostaglandin (PG) E(2) gel with expectant management for women with an unfavorable cervix who wish to attempt a vaginal birth after cesarean section.Study Design: This outpatient study was a randomized, multicenter investigation involving pregnant women at term with one previous low transverse cesarean section. Each had an unfavorable cervix (Bishop score 35% and is an important consideration when evaluating dysfunctional labor. PMID- 10838348 TI - The frequency of extramembranous placement of intrauterine pressure catheters. AB - Objective: To determine, and confirm, the frequency of extramembranous placement of intrauterine pressure catheters.Methods: It is estimated that 15% of all deliveries in the United States (600,000 deliveries/year) receive intrauterine pressure catheters (IUPCs). This study quantifies the frequency of inadvertently placing IUPCs between the chorioamnionic membranes and the myometrium. An attempt was made to aspirate amniotic fluid during placement. If amniotic fluid was obtained, the catheter was considered to be within the amniotic cavity. If amniotic fluid was not obtained but the catheter appeared to be clinically working (possibly extramembranous), normal clinical management was continued until just before the catheter was removed. At that time, 2 mL of indigo carmine was injected. The chorioamnionic membrane was examined to confirm the location of dye (intraamnionic vs extramembranous).Results: Approximately 40 Koala (Clinical Innovations, Inc.) and 40 Intran Plus catheters (Utah Medical Products, Inc.) were placed. In 70% of all first insertions, amniotic fluid was not obtained. When multiple attempts (2-5) were made, amniotic fluid was obtained. There was no statistically significant difference between IUPC brands. Dye confirmed extramembraneous placement.Conclusions: IUPCs are frequently (inadvertently) placed extramembraneous. Upon first insertion it is possible that as many as 30% of IUPCs do not go into the amniotic space. If fluid is not obtained upon placement, a different location should be tried since if amniotic fluid is visualized in the catheter the percentage of intraamnionic placement increases. Adverse results of artefactual waveforms, placental perforation, and abruption with or without amnioinfusion can occur when IUPCs are placed extramembraneous. PMID- 10838349 TI - Transverse arrest: a review of outcomes of rotational forceps and cesarean delivery at a single center. AB - Objective: To assess whether Kielland forceps rotational delivery causes a significant increase in maternal or perinatal morbidity in comparison with cesarean delivery in the clinical situation of transverse arrest. Rotational forceps delivery has been reported to have unacceptably high maternal and neonatal complication rates when compared with either spontaneous vaginal delivery or non-rotational forceps delivery. Clinically, however, we considered the more relevant comparison between Kielland rotational forceps delivery and cesarean delivery as these are generally the two modes of delivery for the fetus in transverse arrest.Study Design: The charts of all the mothers delivered by Kielland forceps or cesarean delivery for the main indication of transverse arrest between July 1995 and June 1996 were reviewed. The following were evaluated: Apgar scores at 1 and 5 minutes, cord pH, birth weight, neonatal superficial facial laceration or bruising, bone or nerve injuries, maternal injuries, 3rd and 4th degree perineal or vaginal lacerations, post-partum fever. Statistical analysis was by chi(2) (for Apgar scores), two-tailed Fisher Exact test (for post-partum fever), and t test (for birth weight and cord pH). A P value <.05 was considered statistically significant.Results: Fifty-two Kielland forceps and 30 cesarean deliveries were performed. One-minute Apgar scores of less than or equal to 7 were noted in 11.5% and 6.7% of the forceps and cesarean section groups, respectively (P = NS); mean arterial cord pH was 7.26 for both groups; no cases of post-partum fever were noted in the forceps versus 16% in the cesarean group (P <.05); the mean weight was 3,316 and 3,569 g for the forceps and cesarean group, respectively (P <.05). The Kielland group was also characterized by the following: 19 facial bruising and 2 temporary facial nerve injury (resolved by the time of discharge), 3 perineal lacerations, and 3 vaginal tears. No blood transfusions were required for either group.Conclusions: To our knowledge, this is the first study that compares rotational forceps delivery for transverse arrest with its true clinical alternative, cesarean delivery.No significant difference in cord pH and Apgar scores were noted between the two groups. The birth weight was greater and post-partum fever was more common in the cesarean section group. Neonatal facial bruising, temporary facial nerve palsy, and maternal perineal and vaginal lacerations were more common in the forceps groups. This study suggests that rotational forceps may be a reasonable alternative to cesarean delivery in the management of transverse arrest. PMID- 10838350 TI - Outcome predictors for patients delivered by cesarean section for fetal distress. AB - Objective: To determine predictive variables identifying infants admitted to neonatal intensive care (NIC) following cesarean section for fetal distress in labor at term.Methods: Two hundred eight patients were studied. Sixty-six patients delivered by cesarean section at term for fetal distress were compared to 142 term patients not diagnosed as fetal distress. The outcome indicator was admission to NIC. Patients diagnosed as fetal distress were studied to determine variables that increased prediction of adverse outcome. Variables studied were patient age, induction of labor, augmentation of labor, epidural anesthesia, birth weight, antepartum complications, and intrapartum complications. Comparisons were by Fishers Exact text and logistic regression.Results: Twenty six infants were admitted to NIC. Eleven had a diagnosis of fetal distress and 21 had a diagnosis of antepartum complications. Fetal distress was not associated with admission to NIC (P =.26) and had a low sensitivity (42%) and positive predictive value (17%). Antepartum complications, intrapartum complications, and birth weight were associated with admission to NIC (P =.00001) (P =.04) (P =.05). Antepartum complications had a sensitivity of 81% and a positive predictive value of 33% for admission to NIC. The presence of both fetal distress and antepartum complications increased the positive predictive value to 91%. Only one infant was admitted to NIC with a diagnosis of fetal distress without antepartum complications. Positive predictive value 2.4%, negative predictive value 96%. Birth weight when dichotomized at the 5th percentile (2,606 g) had a sensitivity of 20% and a positive predictive value of 50% for admission to NIC. Five of 10 infants with a birth weight below the 5% percentile were admitted to NIC.Conclusion: Antepartum complications coupled with fetal distress in labor are a strong predictor of adverse outcome, which is not altered by cesarean section. There are two groups of patients with fetal distress in labor at term, and the group with antepartum complications or subtle reduction in birth weight are at extreme risk for adverse outcome. It appears that cesarean section for fetal distress may rescue infants without antepartum problems but may not rescue those with a challenging antepartum course, lending credence to the feeling that fetal distress in labor may be an indicator of prior stress in many patients. PMID- 10838351 TI - Elective cesarean hysterectomy vs elective cesarean section followed by remote hysterectomy: reassessing the risks. AB - Objective: The purpose of this study was to compare the risks of elective cesarean hysterectomy with the risks of elective cesarean section followed by remote hysterectomy.Methods: A census of elective cesarean hysterectomies (n = 31) and a random sample of 200 cesarean sections and 200 hysterectomies performed by the authors between 1987 and 1996 were evaluated. Only elective repeat and primary cesarean section patients without labor were selected for study (n = 86). Total abdominal hysterectomies were drawn from the sample (n = 60), excluding cancer cases, patients over 50 years old, and those with ancillary procedures other than adnexectomy and lysis of adhesions. General probability theory was used to calculate a predicted complication rate of cesarean section followed by TAH from the complication rates of the component procedures done independently. This predicted combined complication rate was then compared to the observed rate of complications from cesarean hysterectomy to evaluate the risks of the two alternative treatment regimens.Results: Elective cesarean section and total abdominal hysterectomy had complication rates of 12.8% and 13.4%, respectively. The predicted combined complication rate for elective cesarean section followed by TAH was 24.5%. The observed rate of complications for elective cesarean hysterectomy was much lower (16.1%). Although bleeding complications were similar for the two regimens, the rate of transfusion was higher for cesarean hysterectomy (13.0%) than for cesarean section (0%) and TAH (3.4%) alone. Eighty percent of the cesarean hysterectomy patients would have been candidates for autologous blood donation, had it been available.Conclusions: Elective cesarean hysterectomy has a lower risk of complications than elective cesarean section followed by remote abdominal hysterectomy and should be preferred. Transfusion risks are higher for cesarean hysterectomy but can be decreased by the use of autologous blood. PMID- 10838352 TI - Cesarean hysterectomy for placenta previa accreta, increta, and percreta: a strategic approach to minimize surgical hemorrhage. AB - Objective: A new approach to cesarean hysterectomy aimed at minimizing surgical hemorrhage in difficult cases of placenta previa accreta is introduced.Methods: Diagnoses of placenta percreta with bladder invasion, and of placenta increta, respectively, were made by magnetic resonance imaging in two third-trimester patients with multiple previous cesarean deliveries and symptomatic placenta previa. Elective cesarean hysterectomy was planned in both cases. A novel approach to mobilization of the bladder and lower uterine segment at cesarean hysterectomy was applied in both cases.Results: Cesarean hysterectomy by a modified approach was accomplished uneventfully in both cases. Blood loss was 900 mL and 1,100 mL respectively. Neither patient required blood component therapy.Conclusions: A novel approach to cesarean hysterectomy result was associated with limited blood loss and without the need for blood component therapy in two patients at risk for surgical hemorrhage. PMID- 10838353 TI - Vaginal birth after cesarean delivery: a group practice's approach to minimizing failed trial of labor. AB - Objective: Among women attempting a trial of labor (TOL) after a prior abdominal delivery, 60-80% accomplish a vaginal birth after cesarean (VBAC). McMahon and coworkers (N Engl J Med, 1996) have indicated that at a 60% success level for TOL, the remaining 40% incurred enough major complications that the scheduled repeat cesarean section group was less morbid overall. The same authors speculated that a success rate of 80% might be necessary for the TOL group's morbidity to be superior. We sought to review our group's patient selection experience during an interval when successful TOL consistently exceeded 80%.Methods: The study interval ranged from January 1995 through June 1997 and was limited to patients with one previous low transverse cesarean section. Rather than using administrative or charge-related diagnoses, we analyzed a departmental database that included each delivering physician's selection of one of four VBAC categories: successful VBAC, unsuccessful VBAC, patient declined trial of labor, or physician advised against trial of labor. All deliveries were at a single institution and were performed by one of seven obstetricians in a group practice.Results: During the study interval, 332 women provided a history of a single previous cesarean delivery. Of these 332, a total of 173 attempted a TOL and 150 of the 173 (87%) were successful. Fifty-eight of the 332 (18%) declined a trial of labor despite being assessed as excellent candidates, and 101 (30%) were advised against a TOL by their physician. Most common reasons for physicians discouraging labor included malpresentation, fetal macrosomia, and clinically small pelvis. Complications for the 23 of 173 (13%) experiencing a failed TOL included 1 asymptomatic partial separation of a uterine scar and 4 cases of puerperal fever; neither transfusion nor hysterectomy was required.Conclusions: This study demonstrates that in a population of women with one prior cesarean delivery, it is possible for a group practice to achieve a TOL success rate exceeding 80%. In our opinion the managing physician selecting out those patients least likely to attain a successful TOL contributes to a low failed TOL rate. We speculate that an analysis for best practice patterns within our group might reveal information of value for future practice guidelines. PMID- 10838354 TI - Elective repeat cesarean delivery vs trial of labor: a comparison of morbidities in a community hospital setting. AB - Objective: In an attempt to reduce the cesarean delivery rate nationally, many obstetricians are offering a trial of labor to their patients who have had a prior low transverse cesarean delivery. Many studies have demonstrated the success and safety of a vaginal birth after cesarean. However, few studies have actually compared elective repeat cesareans with a trial of labor. Recent articles suggest that the morbidities associated with a failed trial of labor (TOL) may be more serious than that associated with an elective repeat cesarean delivery (RC/D). Our objective was to review and compare the morbidities and complications of repeat elective cesarean delivery versus patients attempting a trial of labor in our institution.Methods and Materials: A retrospective or nonconcurrent cohort study was conducted. Inclusion criteria included all women at our institution who had delivered from July 1993 through March 1997 and had a prior cesarean delivery. Patients were eligible for a trial of labor according to the recommendations of ACOG Practice Patterns. Exclusion criteria were nonvertex presentation, prior classical or T-incision, placenta previa, previous myomectomy, or multiple gestation. The patients were divided into those who underwent an elective repeat delivery and those who consented to a trial of labor. Outcomes studied included major complications (uterine rupture or operative injury) and minor complications (puerperal fever, postpartum hemorrhage requiring transfusion or operative intervention, or abdominal wound infection). Data were ascertained from medical records, the delivery log book, and the Quality Improvement data base. Prior to performing this study, a power analysis was conducted using an alpha of 0.05 and beta of 0.20. The required number in each arm was 2,280 to determine a 50% difference in outcome.Results: There were 1,148 women who had a repeat cesarean delivery during the study time period. Of that, 174 were excluded by the criteria, leaving 973 eligible. There were 1,030 women who underwent a trial of labor. Seven hundred seventy-three women successfully delivered vaginally (75%). The overall maternal morbidity was 2.4%; 0.8% had a major complication and 1.5% had a minor complication. The total complication rate was similar for both the trial of labor group (TOL) and the elective repeat cesarean delivery group (RC/D): 2.33% and 2.57%, respectively. However, major complications were more frequent among those that attempted a trial of labor than those who underwent an elective repeat cesarean delivery; 1.65% versus 0.1% (relative risk 16.08 with a 95% CI of 2.14-120.57; P =.00024). The relative risk for minor complications in the TOL group compared to the RC/D group was 0.28 with a 95% CI 0.12-0.64 (P =.001). Stratified analysis of the TOL group was performed. Compared to the RC/D group, those who failed VBAC had a relative risk for major complications of 56.85 using a 95% CI of 7.45-428.37. The successful VBAC group did not have statistically higher major complication rate (RR = 2.52, CI 0.23-27.74, P =.4).Conclusions: At our institution, there is a trend that major maternal complications are more frequent among those who have a trial of labor compared to those who have an elective repeat cesarean delivery. This trend was even more evident in those who failed a VBAC trial. When VBAC was successful, complications did not seem to be higher than repeat cesarean delivery. Our study did not have the power because of insufficient numbers in each arm. However, data collection is continuing to achieve the power to this study. This prompted us to reconsider the safety for a trial of labor. Perhaps more stringent criteria for selection of VBAC candidates and determination of when a trial of labor should be abandoned need to be defined. Ongoing study is warranted in this area. PMID- 10838355 TI - Home subcutaneous metochlopramide therapy for hyperemesis gravidarum. AB - Objective: To evaluate the efficacy, safety, and economics of home subcutaneous metochlopramide (Reglan) therapy for hyperemesis gravidarum.Methods: All charts of patients who underwent home therapy via a standard protocol utilizing a pump to administer metochlopramide subcutaneously between January 1994 and December 1995 were reviewed. Data were collected on the resolution of symptoms, reason for discontinuance of therapy, additional antiemetic therapy before, during, and after subcutaneous metochlopramide therapy, and side effects.Results: A total of 301 patients received home subcutaneous therapy with metochlopramide: 195 (64.8%) patients had complete resolution of symptoms, 32 (10.6%) patients discontinued therapy because of side effects, and 43 (14.3%) discontinued therapy because of worsening symptoms. A total of 16 (5.2%) patients discontinued therapy for other/nonspecific reasons. Two hundred twenty-two (73.8%) patients received one or more antiemetics prior to subcutaneous metochlopramide therapy. This number decreased to 6 (3%) of the group successfully treated who required any maintenance antiemetic after therapy was completed. One hundred sixty-four (54.5%) patients experienced some side effects. The vast majority of these were mild and did not require discontinuation of therapy. Eleven patients experienced extra pyramidal symptoms, 9 of which were discontinued therapy. The average daily costs for each home therapy patient was $265.00/day. This compares to the average daily costs of $1,370.00 for inpatient therapy for the same Diagnosis Related Group (DRG) in a Midwest five-hospital system.Conclusion: Home subcutaneous metochlopramide therapy appears to be a safe and effective treatment for hyperemesis gravidarum. The home setting results in decreased costs compared to inpatient hospitalization and may constitute a therapeutic, less stressful environment. PMID- 10838356 TI - Effects of gestational and overt diabetes on placental cytochromes P450 and glutathione S-transferase. AB - Objective: Animal and in vivo human studies have observed that diabetes alters the expression of hepatic metabolizing cytochrome P450 (CYP) and glutathione S transferase (GST) enzymes. The placenta has the ability to metabolize a number of xenobiotic and endogenous compounds by processes similar to those seen in the liver. Our objective was to compare placental xenobiotic metabolizing activity in diabetics to matched non-diabetic controls to determine if the presence of diabetes alters placental xenobiotic metabolizing activity.Methods: The catalytic activities of 7-ethoxyresorufin-O-deethylation [EROD] (CYP1A1), chlorzoxazone 6 hydroxylation (CYP2E1), dextromethorphan N-demethylation (CYP3A4), dextromethorphan O-demethylation (CYP2D6), and 1-chloro-2,4-dinitrobenzene (CDNB) conjugation with glutathione (GST) from placentas of diet controlled (class A1) and insulin-dependent (class A2) gestational diabetics and overt diabetics were compared to matched controls.Results: No differences in EROD activity were observed among overt or gestational diabetics and their respectively matched controls. CYP2E1, 2D6, and 3A4 enzyme activity were not detected in human placentas. In contrast, GST activity was significantly reduced by 30% (P <.05) in overt diabetics as compared to their matched controls and gestational diabetics.Conclusion: Pregnant women with overt diabetes have reduced GST activity in the placenta, which could potentially result in exposure of the fetus to harmful reactive electrophilic metabolites. PMID- 10838357 TI - Combining human placental lactogen with routine glucose challenge tests. AB - Objective: To test the hypothesis that among gravidas with abnormal 1 hour glucose challenge tests (GCT), human placental lactogen (HPL) is higher for those found to have gestational diabetes.Study Design: Coincident with the GCT, serum was used to measure HPL by radioimmunoassay technique. Interassay and intraassay coefficients of variance were 6.0% and 1.4%, respectively. All GCT > 130 mg% were followed by a 100 g oral GTT (Carpenter criteria).Results: Two hundred fifty seven women whose mean age was 29.4 years were studied. Twenty-two percent (57/257) had an elevated GCT. Forty-nine percent (25/57) of those with a positive screen were found to have an abnormal GTT. The mean HPL for this group of 25 women with gestational diabetes was 5.85 +/- 2.55 mg/mL and was significantly higher than the 3.38 +/- 1.40 mg/mL level in the 32 (32/57) women who had an abnormal GCT but a normal GTT, P =.034. The mean HPL for women who had a normal GCT screen was 4.68 +/- 1.64 mg/mL and was not statistically different from the HPL in women with an abnormal GCT but who had a normal GTT, P =.1. However, among the 11 women with a normal GCT but whose infants weighed >4000 g, the mean HPL was 5.83 +/- 1.29 mg/mL, which was similar to the mean HPL found in women with gestational diabetes.Conclusion: Our findings suggest that HPL levels are lower in groups of women with positive GCT but a negative GTT when compared to groups with a positive GCT but an abnormal GTT. Adjuvant HPL measurements may improve the positive predictive value of GCT. PMID- 10838358 TI - Screening for gestational diabetes mellitus in adolescent Hispanics. AB - Objective: To determine the incidence of gestational diabetes mellitus in an adolescent Hispanic population and calculate the cost of screening.Methods: A retrospective chart review of deliveries to adolescent (<20 years old) Hispanic women from January 1991 through May 1996 was performed. The incidence of gestational diabetes mellitus (GDM) was determined and the cost of screening calculated.Results: Three hundred twenty-six adolescent Hispanic women were screened for GDM with a 1-hour 50-g oral glucose tolerance test at an average gestational age of 26 weeks (range 11-39 weeks), at a cost of $26 per test. Thirty (9.2%) had positive screens defined as a plasma glucose level of >/=140 mg%. Three-hour 100-g oral glucose tolerance tests were performed on those with positive screens, at a cost of $124 per test. Five women were diagnosed with GDM, an incidence of 1.5%. The cost to diagnose each case of GDM was $2,569.Conclusion: The incidence of GDM in this population is low, and therefore universal screening of Hispanic adolescents may be unwarranted and cost prohibitive. Large prospective trials evaluating maternal and neonatal outcomes would provide the best means to assess the value of screening. PMID- 10838359 TI - Monitoring birth outcomes in the Sumatriptan Pregnancy Registry. AB - Background: Health professionals often need to make medical management decisions by weighing potential risk versus benefit. Because 50% of pregnancies in the United States are unplanned, the likelihood for unintentional first-trimester exposure to medications is high and increases when women of childbearing potential (ages 15-44) are taking medications used to treat chronic or recurrent conditions, such as migraine. Prospective collection of pregnancy exposure and outcome data is one method for providing data to the medical community as a tool for making risk assessments.Methods: The Sumatriptan Pregnancy Registry is an international, exposure-registration and follow-up study to prospectively collect prenatal sumatriptan exposure and outcome data. An advisory committee reviews data and assists in disseminating information. Committee members include independent scientists with expertise in obstetrics, pediatrics, neurology, internal medicine, epidemiology, clinical genetics, and teratology.Results: As of April 30, 1997, 181 outcomes were prospectively reported from pregnancies involving use of sumatriptan. Of the 171 outcomes where earliest exposure was in the first trimester there were 143 infants without birth defects, 4 live infants with birth defects, 1 stillbirth with birth defects [12 spontaneous pregnancy losses, 2 stillbirths, and 9 induced abortions without birth defects]. The proportion of outcomes involving birth defects, 5/148, 3.4% (95% CI 1.3%, 8.1%), does not differ from the expected proportion in the general population.Conclusions: To date, no pattern has emerged in either the prospectively or retrospectively reported defects. Registry data are disseminated to interested health care providers who are then in a more informed position when making medical management decisions. However, more data are needed before definitive conclusions can be drawn about the safety of sumatriptan use in pregnancy. PMID- 10838360 TI - Monitoring pregnancy outcomes following prenatal drug exposure through prospective pregnancy registries and passive surveillance: a pharmaceutical company commitment. AB - Objectives: Glaxo Wellcome (G.W.) becomes aware of prenatal exposures to its medications from as early as the clinical trial phase of development. An international process for monitoring prenatal exposure to all G.W. medicines has been developed utilizing a passive surveillance system and, for specific products, pregnancy registries. Additionally, G.W. jointly sponsors the multi company Antiretroviral and North American Antiepileptic Drug (AED) Pregnancy Registries.Study Design: The registries are observational, case-registration and follow-up studies designed to detect evidence of teratogenicity associated with specific medications. Pregnancies are registered prospectively following prenatal exposure to the registry medication. An advisory committee for each registry reviews data and assists in dissemination of information. Committee members include independent scientists with expertise in fields such as obstetrics, teratology, epidemiology, pediatrics, and the relevant therapeutic areas.Results: The following data are from the prospective first-trimester exposures in each registry. Through December 1996, the proportion of outcomes in the Acyclovir Pregnancy Registry with birth defects (n = 17/505) is 3.4% (95% CI 2.0%, 5.4%). Through March 1997, the proportion of outcomes in the Lamotrigine Pregnancy Registry with birth defects (n = 4/76) is 5.3% (95% CI 1.7%, 13.6%). Through April 1997, the proportion of outcomes in the Sumatriptan Pregnancy Registry with birth defects (n = 5/148) is 3.4% (95% CI 1.3%, 8.1%). The newer Valacyclovir and Bupropion Pregnancy Registries have insufficient data for analysis. None of the registries have provided a risk estimate exceeding that expected in the general population, and no pattern of defects has been observed.Conclusions: The outcomes accumulated to date represent a sample of insufficient size for reaching conclusions regarding the possible teratogenic risk of using these drugs in pregnancy. Data obtained through these registries are shared with the medical community as a supplement to animal toxicology studies and to assist in weighing potential risks and benefits of treatment for individual patients. The success of the registries depends on the continued willingness of the OB/GYN community to notify the registries of prenatal exposures. PMID- 10838361 TI - Acute fatty liver in the second trimester of pregnancy. AB - Objective: Acute fatty liver of pregnancy (AFLP) is an uncommon, potentially fatal disorder that usually occurs in the late third trimester of pregnancy. We present the first reported case of acute fatty liver in the second trimester of pregnancy.Methods: We report the clinical and laboratory findings in a patient with AFLP who presented in the second trimester of pregnancy.Results: A 37-year old G5P4 woman presented at 22 weeks gestation (by 18 weeks ultrasound) with nausea and vomiting. She was normotensive, had no proteinuria, had elevated SGOT and SGPT (266 and 261, respectively), negative hepatitis studies and a normal platelet count. She was managed conservatively for presumed cholelithiasis until 24 weeks gestation when she was transferred to our facility because of worsening SGPT and SGPT (368 and 505, respectively), jaundice (total bilirubin of 8.9 mg/dL), hypoglycemia, and laboratory evidence of disseminated intravascular coagulation (DIC) (PT = 18.6, PTT = 56, hypofibrinogenemia and presence of fibrin split products). Ultrasound showed singleton fetus (EFW 450 g) with total placenta previa. Computed tomography scan of the abdomen revealed decreased hepatic density consistent with AFLP. Delivery of a nonviable fetus was effected after transfusion of fresh frozen plasma. Postoperatively, the patient had rapid resolution of DIC, jaundice, and hypoglycemia; liver transaminases normalized 5 days postoperatively and the patient was discharged home in good condition 5 days later.Conclusion: It has been traditionally stated that AFLP occurs in the late third trimester of pregnancy. This case demonstrates that, even in the second trimester of pregnancy, the diagnosis of AFLP should be considered as a cause of deteriorating liver function, jaundice, and DIC. PMID- 10838362 TI - Acute fatty liver of pregnancy: an experience in diagnosis and management of eight cases. AB - Objective: To investigate the early recognition and management of acute fatty liver of pregnancy (AFLP) to improve the maternal and fetal survival.Study Design: Eight cases presenting with AFLP managed in our hospital during the past 212 years were studied retrospectively with emphasis on presenting symptoms, laboratory findings, and the time for liver biopsy. Also, to report the maternal and fetus outcomes in such cases.Result: The mean gestational age at onset was 34 +/- 2 weeks (range 30-37 weeks). All cases were primigravida. In the early stages, all presented with malaise, nausea, vomiting, and epigastric distress followed by jaundice in the third trimester of pregnancy. Three of eight presented with polyuria and polydipsia. Laboratory findings: all had raised transaminases and serum bilirubin (2.9-29.9 mg/dL), hypoalbuminemia (22.4-30 g/L), hypofibriogenemia (< 180 mg/dL), prolonged prothrombin time, and prolonged partial thromboplastin time. Maternal complication was frequent, including hepatic encephalopathy (6), ascites (6), hypoglycemia (5), hematemesis (2), postpartum hemorrhage (5), and preeclampsia (4). Cesarean was performed in 3 cases. One mother died of fulminant hepatic failure, the other cases were survival. There were no fetal deaths. Liver biopsy was done in 8 cases. It is suggested that percutaneous liver biopsy should not be done until the coagulation tests become normal, the amounts of ascites decrease and platelet counts increase after delivery.Conclusion: With increasing awareness, especially in the early recognition of AFLP cases and prompt progressive management, including early termination of pregnancy, and using large-dose infusion of fresh frozen plasma or albumine alternatively, the prognosis of AFLP is obviously improved. PMID- 10838363 TI - Abdominal pregnancy: to remove or not to remove the placenta. AB - Objective: The deleterious effect of abdominal pregnancy on the mother and fetus is in part related to the morbidity of the surgical interventions utilized in its treatment. The purpose of this study is to review outcome in abdominal pregnancy after surgical intervention.Study Design: Charts of patients diagnosed with abdominal pregnancy at our institution between 1984 and 1997 were reviewed. The identified cases were categorized as group I, placenta removed at surgery (n = 10), and group II, placenta left in situ (n = 4). Gestational age, maternal death, duration of hospital stay, blood transfusions, organ excisions, and postoperative readmissions were recorded. Student t test was used for statistical analysis with a P <.05 being significant.Results: Fourteen cases were identified ranging from 7 to 36 weeks of gestation. The diagnosis was made before laparotomy in 6 patients by imaging studies. There were no maternal deaths. Among the 9 in whom placenta was removed, 2 had salpingo-oophorectomy (S-O), 4 had total abdominal hysterectomy and bilateral S-O, and 5 received blood transfusions. One developed DIC requiring massive transfusion after a 7-week placenta was excised from the mesentery. This patient was hospitalized postoperatively for 5 months. In contrast, the 4 patients in whom the placenta was left in situ had neither blood transfusions nor removal of pelvic organs. Their hospital stay was shorter, group II, mean 9 +/- 6 days versus group I, 34 +/- 64 days, P =.0007. This difference was accounted for by the one prolonged hospitalization in group I. No patients in either group were readmitted.Conclusion: The diagnosis of abdominal pregnancy is often not made until laparotomy. Regardless of gestational age, placental excision can cause hemorrhage. Leaving the placenta in situ is potentially less costly and less morbid, and appears to shorten operative time and hospital stay while lowering risk of blood transfusion and of surgical menopause. PMID- 10838364 TI - Comparison of the accuracy of gestational age estimation from fetal heart rate and crown-rump length. AB - Objective: To compare the accuracy of gestational age (GA) estimation predicted by fetal heart rate (FHR) versus crown-rump length (CRL) measurements in early pregnancies.Methods: A total of 145 infertile women with singleton intrauterine pregnancies conceived through in vitro fertilization underwent vaginal ultrasonographic examination in early stage of pregnancy. We performed 197 FHR and 398 CRL measurements. The relationship between GA (counted with the day of oocyte retrieval used as day 14) and the FHR or CRL was explored with regression analysis by means of least squares. Polynomials up to the third order were analyzed, and the coefficient of determination (R(2)) was calculated.Results: Fetal cardiac activity could be detected as early as 38 days, gradually increased to 188 beats per minute at 62 days of gestation. Regression analysis with GA as dependent variable resulted in R(2) values of 0.906, 0.907, and 0.908 for the first, second, and third order polynomials, respectively. The standard deviations around the regression line were 2.90, 2.81, and 2.84 days, respectively. The second order polynomial to describe GA (days) estimated by FHR (beats per min): GA = 3.02E-4*FHR(2) + 1.60E-1*FHR + 2.22E+1. On the other hand, the CRL was measured at 38 days to 62 days of gestation. Regression analysis resulted in R(2) values of 0.827, 0.839, and 0.841, respectively. The deviations were 4.11, 3.83, and 3.89 days, respectively. The second order polynomial to describe GA estimated by CRL (mm): GA = -2.10E-2*CRL(2) + 1.49E+0*CRL + 3.86E+1. There was no difference between the R(2) values and no difference between the standard deviations around the regression lines.Conclusion: Our results indicate no statistically significant difference in the accuracy of GA estimation predicted by FHR versus CRL measurements. But better mathematical fits were obtained when the data of FHR were used for GA estimation. These findings suggest that the FHR measurement may be a new method for early ultrasound dating. This study mentions for the first time the possibility of "FHR dating" with functional data. PMID- 10838365 TI - Correlation of maternal triple screen values with fetal liver length. AB - Objective: To determine if a relationship exists between fetal liver length and any of the three serum markers (AFP, uE3, and hCG) that constitute the maternal triple marker screen.Methods: From March to November of 1996, 124 randomly selected women between 17.0 and 20.3 weeks gestation underwent serum triple screen testing on the same day that an anatomy sonogram was performed by a single sonographer. Fetal liver size was measured between the right hemidiaphragm and the tip of the right hepatic lobe. All triple marker screen results were obtained from the same commercial lab. Utilizing linear regression analysis, liver length adjusted for gestational age was plotted versus AFP, uE3, and hCG measurements (expressed as multiples of the median).Results: Of the 124 subjects, 17 (14%) were white and 97 (78%) were black; the remaining 10 (8%) were unknown or other. Neither hCG nor AFP measurements had a statistically significant association with liver length after taking into account gestational age (linear regression: R = 0.018, P =.848 and R = 0.100, P =.269 for hCG and AFP, respectively). Interestingly, a statistically significant correlation was identified between uE3 levels and fetal liver length (R = 0.205, P =.023).Conclusions: No statistical significance was identified between either maternal serum hCG or AFP and fetal liver size. A statistically significant correlation between maternal serum uE3 and liver size was observed. Although preliminary, sonographic evaluation of fetal liver size has little role in elucidating the etiology of abnormal hCG or AFP levels, but may be relevant in the interpretation of abnormal uE3 values. PMID- 10838366 TI - Accurate prediction of term birth weight from prospectively measurable maternal characteristics. AB - Objective: To determine whether accurate prediction of term birth weight is possible based on maternal characteristics routinely measured remote from term, and to compare this technique to more expensive methods requiring obstetrical ultrasound examinations.Methods: Two hundred fifty-nine normal, non-smoking, non diabetic Caucasian gravidas with uncomplicated term gestations were studied. Seven maternal characteristics (age, parity, height, weight, level of obesity, rate of pregnancy weight gain, and glucose screening test result) and two fetal characteristics (fetal gender and length of gestation) were evaluated alone and in combination for their predictive values in determining birth weight. A cross validated split-sample multiple regression analysis was used to determine which combinations of these variables were significant and a birth weight prediction equation was developed. Predictive accuracy was assessed using a jackknifing procedure, and results were compared to similar types of birthweight predictions obtained both via previous algorithms based upon maternal characteristics and those developed for use with fetal ultrasonographic biometric data.Results: Significant predictors of term birth weight were gestational age, fetal gender, parity, maternal height, maternal weight, and third trimester maternal weight gain rate. These prospectively measurable variables explained 33% of the variance in birth weight and predicted birth weight to within 10.8%. These results were compared to those obtained from other previously published algorithms and were more accurate than all others derived from either maternal characteristics or fetal ultrasonographic data. Our term birth weight prediction equation is: birth weight (g)=gestational age (days)x[9.40+0.255xgender+ 0.000232xheight (cm)xmaternal weight at 26 wk (kg)+ 4.89x3rd trimester weight gain rate (kg/d)x(parity+1)]where: gender=-1 for females;+1 for males;0 for unknown gender gestational age=conceptual age (days)+14Conclusion: Term birth weight can be accurately predicted using routinely measurable maternal characteristics. Birth weight estimates using our equation are both prospectively derivable starting from the end of the second trimester and more accurate than any previously devised algorithms, including methods based upon ultrasonographic fetal biometric data. PMID- 10838367 TI - Hysterosonography as an adjunct to transvaginal sonography in the evaluation of intraluminal lesions of the uterine cavity. AB - Objective: Hysteroscopy is a simple technique that can be used to outline the contours of the uterine cavity under real-time ultrasound guidance. This study was undertaken to determine whether hysterosonography can be used as an adjunct to transvaginal sonography in predicting either the presence or absence of intraluminal lesions in the uterine cavity and in distinguishing these lesions from subendometrial or myometrial lesions.Methods: In this retrospective analysis, 1,467 transvaginal sonograms performed in the office of a single gynecologic oncology practice from March 1996 through June 1997 were reviewed. Of these, 38 sonograms were considered to have possible intraluminal lesions of the uterine cavity and thus further evaluated by hysterosonography. The principle investigator performed all the hysterosonograms by infusing 15-30 mL of sterile saline directly into the uterine cavity using a Soules 5.3F transcervical catheter. Real-time transvaginal sonography was performed simultaneously, and representative images were compared with the initial transvaginal sonograms.Results: Successful hysterosonograms were obtained in 36 patients (97.3%). One patient had cervical stenosis from a previous conization, precluding access to the uterine cavity. The hysterosonography was tolerated well by the patients with minimal discomfort. There was no incidence of endometritis. The median age was 56.2 years. Postmenopausal bleeding was the most common presenting symptom (30.0%). The indications for hysterosonography were abnormal findings on the initial transvaginal sonograms including lesions suggestive of intraluminal mass (73.0%), thickened endometrial stripe (13.5%), and increased echogenicity (10.8%). Hysterosonography revealed presence of intraluminal masses in only 11 of the 31 cases (35.5%) that had been suggested to contain either a mass or thickened endometrial stripe. Subsequent hysteroscopy, dilatation, and curettage confirmed the presence of endometrial polyp in all 11 cases. The remaining 65.5% contained no intraluminal masses or thickened endometrial stripe.Conclusion: Hysterosonography can be a valuable adjunct to transvaginal sonography and may be used in selected cases to eliminate the need for hysteroscopy, dilatation, and curettage. PMID- 10838368 TI - Experience with collaborative management of symptomatic fibroids by gynecology and interventional radiology. AB - Objective: The purpose of this paper is to review the initial collaborative experience of a gynecologist and an interventional radiologist in the utilization of embolotherapy in the management of symptomatic uterine myomas.Methods: One hundred five consecutive subjects treated by embolotherapy were selected for review. All patients derived from a single gynecologic practice and were evaluated by the same examiner, using standardized criteria. Embolotherapy was carried out in all subjects by the same interventional radiologist using 500-700 um polyvinyl alcohol particles injected bilaterally into the uterine arteries utilizing uniform technique and endpoints.Results: Size reduction of 20-85% confirmed by follow-up ultrasound was achieved. Ninety percent of patients noted improvement in their overall symptoms. Eighty percent of patients were able to resume normal activity 4 days after therapy, the remainder of 20% within 2 weeks. One patient subsequently underwent hysterectomy. At 1-year follow-up, changes in size and menorrhagia persisted.Conclusion: The collaborative treatment of fibroids by gynecologists and interventional radiologists appears to offer an effective alternative to women with symptomatic uterine fibroids. PMID- 10838369 TI - Comparison of bromfenac and naproxen sodium in the management of primary dysmenorrhea. AB - Objective: To compare a new analgesic with an established standard and placebo in the treatment of primary dysmenorrhea.Methods: Bromfenac (B) 25 or 50 mg prn, up to 4 doses daily, was compared with naproxen sodium (N) (550/275 mg) and placebo (P) in a double-blind crossover randomized trial where 54 women were treated over 4 menstrual periods. Each treatment lasted up to 3 days. The first-dose analysis was based on a 6-hour observation period. Total pain relief (TOPAR) and summed pain intensity difference (SPID) scores were calculated from hourly collected data as areas under the respective time curves.Results: All 3 active treatments were superior to placebo in all summary variables (SPID and TOPAR for 3 and 6 hours) and corresponding peak pain relief and peak PID, duration of pain half gone, and global assessment. Both B doses were superior to P in providing significant pain relief between hours 1 and 6, whereas N was superior to P at hours 1, 3, and 4. Good to excellent global rating was achieved by 74% of patients on B 50 mg, 59% on B 25 mg, 57% on N and 33% on P (P <.001) after the first dose. Patients rated the trial medication superior to their usual treatment as follows: B 50 mg: 65%, B 25 mg: 55%, N: 51%, P: 29% (P <.001). Adverse events were infrequent and not significantly different among the 4 treatments.Conclusion: Both doses of bromfenac were significantly superior to placebo and at least as efficacious as naproxen sodium in management of primary dysmenorrhea. PMID- 10838370 TI - Polysomnographic characteristics of sleep disorders in chronic pelvic pain. AB - Objective: We performed a pilot study to determine if patients with chronic pelvic pain have a consistent or unique sleep disorder by a polysomnographic sleep study.Methods: Overnight polysomnography was performed prospectively in 11 women with chronic pelvic pain (ages 33-44 years, mean 35). None of the women were taking sedating or psychoactive medications in the month prior to testing. Sleep indices were derived using polysomnography scoring guidelines currently recommended by the American Sleep Disorder Association.Results: Statistical comparisons with age-matched normative data (Williams and Karacan, 1974) revealed increased stage 1 sleep (P < 10(-7)), reduced REM sleep (P < 10(-6)), increased slow wave sleep (P <.001), and REM sleep onset latencies (P <.01) in women with chronic pelvic pain. One woman had periodic leg movements, but none had obstructive sleep apnea.Conclusion: Previously, sleep disorders in patients with chronic pelvic pain (CPP) have been characterized by history alone. We have characterized sleep disturbances in individuals with CPP by polysomnographic sleep studies. Patients with CPP have different sleep patterns than patients with depression. Our preliminary findings and review of the literature conclude that patients with CPP have a unique sleep disorder. We are currently studying the effects of low doses of amitriptyline in altering sleep patterns to determine if there is a clinical response. PMID- 10838371 TI - A food interaction study of bromfenac, naproxen sodium, and placebo in cesarean section patients. AB - Objective: This double-blind study was to compare the effect of food on the analgesic response to bromfenac, naproxen sodium, and placebo.Methods: Single doses of bromfenac (BRO) 25 mg, naproxen Na (NAP) 550 mg, or placebo (PLA) were given to 284 patients with moderate or severe pain following cesarean section. A standard breakfast was provided for the "fed" patients. "Fasted" patients received no food 3 h before and 1 h after the dose. Treatments were compared over 8 h using standard scales for pain intensity and pain relief. Plasma levels of BRO were measured in 7 fasted and 12 fed patients.Results: BRO and NAP were significantly superior to PLA; food did not affect the response to any treatment: As expected, mean peak plasma levels of bromfenac were reduced by food by about 65%.Conclusion: Food reduces the bioavailability of bromfenac but has no effect on the analgesic response. PMID- 10838372 TI - Effect of lidocaine vs ketorolac tromethamine on postoperative laparoscopy pain control: a randomized study. AB - Objective: To evaluate and compare the effectiveness of intraperitoneal lidocaine and intramuscular ketorolac tromethamine for immediate and delayed postoperative pain management in patients undergoing laparoscopy.Methods: Design-Seventy-four patients were prospectively randomized into four groups: intraperitoneal lidocaine; intramuscular ketorolac; both drugs; neither drug.Setting-Community Teaching Hospital Outpatient Surgery Center.Patients-Private and clinic patients needing operative laparoscopy.Intervention-One hundred milligrams of lidocaine was instilled into the peritoneal cavity just before the laparoscopy was completed. Sixty milligrams of ketorolac tromethamine was administered intramuscularly approximately 30 minutes before laparoscopy was completed.Main Outcome Measurements-The amount of analgesic used, the time required in postanesthesia recovery area, and a pain intensity score were used to evaluate the postoperative pain.Results: Pain intensity scores reported by those women receiving ketorolac and both medications were less than scores of untreated controls. Groups receiving either ketorolac, lidocaine, or both drugs required less morphine postoperatively.Conclusion: Both intraperitoneal lidocaine or intramuscular ketorolac are effective in reducing the perception of pain and the amount of morphine required postoperatively. PMID- 10838373 TI - A randomized, placebo-controlled comparison of EMLA() and dorsal penile nerve block for pain relief during neonatal circumcision. AB - Objective: To evaluate the relative efficacies of eutetic mixture of local anesthetics (EMLA()) cream and dorsal penile nerve block (DPNB) for pain relief during neonatal circumcision.Methods: After parental informed consent, appropriate-for-gestational age, term, healthy newborns were randomized to receive either EMLA cream and placebo saline DPNB or placebo cream and 1% lidocaine DPNB. Placebo and EMLA cream were prepared by the pharmacy and applied by study nurses. Preloaded syringes of saline or 1% lidocaine were also prepared by the pharmacy for the DPNB injections. This ensured blinding of the surgeons and assistants at the time of circumcisions. Circumcisions were performed with the Gomco clamp technique by one of three obstetrics and gynecology residents. Pain levels were assessed by measuring pulse and respiratory rates and determining Brazelton states at each step of the circumcision. Videotapes were made of each procedure to allow later scoring by a single blinded observer for Brazelton states. Brazelton state scores increase with increased infant pain and distress, as do heart and respiratory rates. Means were compared by t test and proportions by chi(2). A P value of.05 was considered statistically significant.Results: A total of 60 infants were randomized, 29 to DPNB and 31 to EMLA. Preoperative Brazelton state scores, heart rates, and respiratory rates were similar in both groups. Brazelton state scores were lower in the DPNB group at each step of the procedure-injection, dissection of the foreskin, clamp placement, and clamp removal-and postoperatively and overall, but the differences only reached statistical significance during dissection and clamp removal. Heart and respiratory rates also were lower at all surgical steps in the DPNB group but only reached statistical significance during postoperative observation.Conclusions: Although both EMLA and DPNB have been shown in other studies to decrease pain during neonatal circumcisions, DPNB has greater effectiveness in relieving the pain of circumcision. PMID- 10838374 TI - Are the long-term adverse effects of laparoscopic presacral neurectomy for the management of central pain associated with endometriosis acceptable? AB - Objective: To assess the long-term genitourinary and gastrointestinal complaints following presacral neurectomy.Design: A prospective postoperative follow-up of patients who underwent laparoscopic presacral neurectomy and treatment of endometriosis.Materials and Methods: The mean follow-up of the 67 women (mean age 27.5 years, range 16-58 years) was an average of 36.8 months with a range of 6-69 years. Main outcome variables include diarrhea, constipation, bladder and urinary complaints, vaginal dryness, dyspareunia, and orgasm. The degree of pain and dysmenorrhea after surgery was also elevated.Results: Diarrhea was reported to have improved after surgery in 39.1% of the patients and none reported any worsening. Constipation improved in 28.6% and worsened in 12.5%. Only one patient suffered from debilitating constipation. Bladder and urinary problems were improved on 25.0% and worsened in 19.2%. A similar proportion of women (19.6%) reported improvement and worsening vaginal dryness. Pain during intercourse improved in 58.9% and worsened in 8.9%. The ability to achieve orgasm improved in 21.6% and worsened in 2.7%. Postoperatively, pain was improved by 80-100% in 46.6% of the women, by 50-80% in 36.5%, by less than 50% in 6.4%, and did not improve in 9.5%. Dysmenorrhea was improved by 80-100% in 35.2% of the women, by 50-80% in 38.8%, by less than 50% in 14.9%, and did not improve in 11.1%. Twelve of 16 patients trying to become pregnant were successful following surgery, two with the aid of in vitro fertilization.Conclusion: After laparoscopic presacral neurectomy, constipation and bladder and urinary problems were reported to have worsened in only a minority of patients. However, diarrhea and dyspareunia improved in a large proportion of patients. Since pelvic pain was relieved by more than 50% in 83.1%, the procedure seems to be associated with an acceptable rate of long-term side effects. PMID- 10838375 TI - Epidural anesthesia performed during labor by obstetricians: outcome analysis. AB - Objective: The objective of this outcome analysis was to determine the frequency, effectiveness, and complications of epidural anesthesia performed during labor by attending obstetricians at Minden Medical Center from January 1993 through December 1995.Methods: Each of the 1,851 obstetrical patient charts for the 3 year time period was reviewed and data were obtained from the 1,565 charts of the patients who received epidural anesthesia in labor.Results: During the 3-year period studied 1,851 patients had obstetrical delivery at Minden Medical Center. There were 147 planned cesareans and 1,704 patients who experienced labor. Among the patients who experienced labor 1,565 (91.8%) received epidural anesthesia performed by their attending obstetrician. Epidural anesthesia was effective for adequate pain relief in 1,484 patients (94.8%). Hypotension requiring ephedrine occurred in 24 patients (1.5%). Subarachnoid puncture requiring a blood patch for treatment of postural headache occurred in 4 patients (0.26%) and inadvertent spinal anesthesia occurred in 3 patients (0.2%). All of the inadvertent spinals were recognized with the test dose of 5 mL of 1.5% lidocaine and no high or total spinals resulted. A high level of anesthesia, above T5 dermatome, occurred in 16 patients (1.0%). No seizures, neurological sequelae, toxicity to the anesthesia agent, anaphylaxis, or deaths occurred. There were no cases of urinary retention secondary to an epidural anesthetic requiring patient use of a retention catheter on discharge from the hospital.Conclusion: Outcome analysis of patients who received epidural anesthesia performed during labor by obstetricians at Minden Medical Center indicated obstetricians at this hospital provided epidural anesthesia for a high percentage (91.8%) of laboring patients effectively (94.8% had adequate pain relief), and with no serious complications. PMID- 10838376 TI - A randomized blinded trial of preemptive local anesthesia in laparoscopy. AB - Objective: Laparoscopy, while routinely performed in the outpatient setting, is associated with considerable postoperative discomfort. Continuing pain experienced after surgery is due to post-traumatic functional changes in both the peripheral nervous system (hyperalgesia) and the central nervous system (hyperexcitability). Local anesthetic infiltrated at time of incision closure has limited effect because hypersensitivity and hyperexcitability have already developed. Preemptive analgesia refers to the blockage of afferent nerve fibers, before painful stimulus, which prevents or reduces subsequent pain even beyond the effect of the block. We tested the hypothesis that local anesthetic administered before skin incision, an example of preemptive analgesia, reduces postoperative pain for women undergoing laparoscopy, as compared to postincisional local anesthetic or placebo.Materials and Methods: Seventy-five patients undergoing laparoscopy for pelvic pain, infertility, or sterilization were randomized to one of three treatment groups. Two 10 mL syringes, labeled "Pre" and "Post," were prepared at time of laparoscopy and contents blinded to anesthesiology, surgeons, and the patient. For treatment group A (preincisional), the presyringe contained 10 mL of 0.5% bupivacaine (50 mg) and the postsyringe contained 10 mL of 0.9% saline. For treatment group B (postincisional) patients, the presyringe contained 10 mL of 0.9% saline and the postsyringe contained 10 mL of 0.5% bupivacaine. For treatment group C (control) patients, both syringes contained 10 mL of 0.9% saline. All patients underwent a standardized general anesthetic induction and maintenance. After the patient was properly positioned and draped, 5 mL of the presyringe was infiltrated into the umbilical incision site. The remaining 5 mL was infiltrated in a similar fashion at the suprapubic trocar placement site. After laparoscopy and immediately prior to closure of the incisions, the postsyringe was infiltrated into both incisions above and below the fascia in a diamond-shaped pattern.For postoperative pain, oral ibuprofen was given, as needed, with 30 mg intramuscular ketorolac tromethamine given if the patient was unable to tolerate oral pain medication. All patients were discharged with 800 mg ibuprofen tablets and asked to take as needed for pain relief. The modified McGill Present Pain Intensity scale was evaluated by nurse interview at 30 minutes, 2 hours, 4 hours, and 24 hours after incision closure. Statistical analysis was accomplished using chi(2) tests for proportional data and ANOVA for pain scores and other parametric data.Results: Fifty-seven patients completed the study protocol. Age, weight, height, race, indication, and operating time did not vary significantly between the three groups. Patients in treatment group A (n = 20) could tolerate a significantly longer time delay to their first analgesic medication. (A: 486.7 +/- 435.3 minutes; B: 229.4 +/- 330.4; C: 143.1 +/- 156.7, P <.001). Their 24-hour pain scores were also significantly lower than either treatment group B (n = 19) or C (n = 18) (A: 0.50 +/- 0.9; B: 1.61 +/- 1.3; C: 1.2 +/- 1.2, P <.02). Although statistical significance was not reached, patients in treatment group A required less total doses of analgesic than either treatment group B or C (A: 2.4 +/- 1.6 doses; B: 3.1 +/- 1.5; C: 3.1 +/- 1.2, P =.07).Conclusions: Preemptive local anesthesia in patients undergoing laparoscopy results in a longer time before analgesic is required and significantly lower pain 24 hours after surgery. PMID- 10838377 TI - A prospective randomized trial of the use of epidural anesthesia during external cephalic version. AB - Objective: To determine if the use of epidural anesthesia improves the success rate of external cephalic version (ECV).Design: Women identified with a singleton fetus in a breech or transverse presentation at a gestational age of >/=37 weeks gestation were offered enrollment in this prospective randomized trial. Inclusion criteria included maternal age of 18 years or older, a non-vertex presentation confirmed by ultrasound, a reactive fetal non-stress test, and an estimated fetal weight of 2,000-4,000 g. A 1:1 randomization was accomplished through a computer generated random numbers table with group assignments sealed in sequentially numbered opaque envelopes. Women in the epidural ECV group had a lumbar epidural catheter inserted through which 2% lidocaine and 100 ug of fentanyl were infused.Results: There were no statistically significant differences between the two groups in gestational age at ECV, placental location, fetal lie, gravidity or parity, estimated fetal weight, or amniotic fluid index. The ECV was successful in 26 of 45 (58%) women with epidural anesthesia compared to 16 of 48 (33%) with no anesthesia (relative risk 1.7, 95% confidence interval 1.1-2.8, P <.05). Fetal bradycardia resulting in discontinuation of the version efforts occurred in two patients in the epidural group and three women in the control population, a non statistically significant difference. There were no maternal complications noted in the study population related to the epidural anesthesia.Conclusions: The use of epidural anesthesia increases the success rate of external cephalic version. PMID- 10838378 TI - The "Femassist": a new device for the treatment of female urinary incontinence. AB - Objective: This study was undertaken in order to determine if the "Femassist" device is a safe and effective treatment for women with the diagnosis of urinary incontinence.Methods: The Femassist is a medical-grade silicon dome-shaped device, worn over the urethra and held securely via suction and a commercially available adhesive lotion. Women with a chief complaint of urinary incontinence responding to local newspaper advertisements were screened for inclusion. Potential candidates underwent medical history, physical (including gynecologic) examination, Papanicolaou test, urine culture and cytology, and multichannel urodynamic testing (including abdominal leak-point pressure measurements). A total of 38 women with documented genuine stress urinary incontinence (GSUI) or mixed incontinence were ultimately recruited into the study and fitted with either the standard or petite-sized Femassist device, according to their individual anatomy. Subjects were assessed before and after 1 month's use. Subjective assessment included quality of life questionnaires, daily voiding and activity diaries, as well as ongoing patient comments retrieved through daily telephone contact with the study nurse. Objective assessment included blinded evaluation of bacteriuria and urinary infection rates and vulvar irritation and ulceration rates.Results: To date, of the 38 women who have completed the study, over 50% reported an improvement in their quality of life including comfort, convenience, and overall satisfaction. In total for all patients studied, the device was worn for a total of 886 days; 82% of these were dry days. Similar results were obtained for women with GSUI and mixed incontinence. Factors associated with successful experience with the device included degree of tissue estrogenization (either naturally or via a topical estrogen preparation), manual dexterity, and degree of motivation. One in five women reported vulvar irritation or urethral discomfort at some point; this was not correlated with percentage of dry days. There have been no reported significant increases in bacteriuria or urinary tract infection rates over patients' baseline experiences.Conclusion: Our preliminary study suggests that the Femassist device is a safe and effective method for the management of female urinary incontinence. PMID- 10838379 TI - The outpatient Closed Burch-M.M. procedure for treatment of genuine stress urinary incontinence with no laparotomy or laparoscopy by newly invented bladder saver device. AB - Objective: To identify a more effective, less invasive surgical technique and tool for treatment of genuine stress urinary incontinence. The new outpatient procedure is one of the most effective ways to treat the patient without sacrificing safety and outcome.Method: The Closed Burch Procedure is a retropubic, trocar and ligature carrier suspension by newly invented bladder saver device that is a major solution to the problem of female genuine stress incontinence. As originally described by Dr. John C. Burch in 1961, the operation entails use of the vaginal wall as an endogenous suburethral sling. The vagina is elevated bilaterally at the urethrovesical junction, thus repositioning the proximal urethra within the abdominal cavity toward Cooper ligament by permanent sutures.Result: After confirmation of diagnosis, 11 patients underwent the new outpatient procedure. The new approach showed no failure of technique or any major complications. It also showed no postoperative urinary retention, no recurrence of stress urinary incontinence up to this time, no wound infection, no hematoma, no rejection of sutures, no nerve damage, no osteitis, and no vaginal enervation.Conclusion: The new outpatient Closed Burch technique is an effective treatment for genuine stress incontinence. The procedure is outpatient with logical anatomical restoration. It is fast and can be done under local anesthesia for some patients.Complications are minimum to none. There is no urinary retention, no self-catheterization, and no major or expensive system needed. The procedure has shown very impressive results. PMID- 10838380 TI - Vaginal paravaginal repair in the surgical treatment of anterior vaginal wall prolapse. AB - Objective: To describe our technique and report our clinical experience with the vaginal paravaginal repair in the surgical treatment of displacement cystocele.Methods: Forty-five patients with bilateral paravaginal support defects underwent vaginal paravaginal repair during a 2-year period at our institution. The technique of vaginal paravaginal repair will be illustrated through the use of anatomic photographs taken at the time of surgery. Demographic data were collected for each patient. Preoperative evaluation, intraoperative parameters, and postoperative course were reviewed.Results: The technique of vaginal paravaginal repair was standardized with transvaginal entrance into the retropubic space bilaterally. The paravaginal defects were identified and repaired using permanent suture in a 3-point closure incorporating the pubocervical fascia, arcus tendineous, and vaginal wall. Concurrent repairs were performed as appropriate.Mean age of the patients was 65.9 +/- 2 years (range 35 76). Thirty-eight patients had advanced prolapse of the anterior vaginal wall beyond the introitus, and 21 patients had coexisting stress incontinence. Postoperatively, the length of stay was 2.6 +/- 1.1 days (range 1-6) and urethral catheterization was required for a median 7 days. Intraoperative complications included 1 case of bilateral ureteral obstruction, and postoperative morbidity included 1 retropubic hematoma requiring re-exploration, 2 vaginal abscesses, and 2 postoperative transfusions.Conclusion: The vaginal paravaginal repair is a safe and effective technique in the surgical correction of vaginal wall prolapse due to a displacement cystocele. The vaginal approach provides adequate exposure to the relevant anatomy and good clinical results. PMID- 10838381 TI - Laparoscopic ureteroureterostomy: a prospective follow-up of 9 patients. AB - Objective: To report the technique and long-term outcome of laparoscopic ureteroureterostomy of injured ureter.Design: A prospective follow-up study.Materials and Methods: Follow-up of nine patients, aged 30-43 years, who acquired intentional ureteral injury during operative laparoscopy for treatment of endometriosis. They were treated between September 1991 and September 1997 by ureteral transection or resection with primary laparoscopic repair by ureteroureterostomy (8 procedures) and ureteroneocystostomy (2 procedures).Results: All operations were successfully completed by operative laparoscopy without need to convert to laparotomy. Estimated blood loss related to the ureteral portion of the procedure was less than 100 mL. Duration of follow up was between 2 months and 6 years. Follow-up intravenous pyelogram revealed complete healing in seven patients. One patient had mild ureteral stricture, which resolved with transvesical ureteral dilatation, and she is doing well 4 years postoperatively. Another patient with severe endometriosis had recurrent ureteral stricture due to fibrosis distal to the anastomosis site. She underwent laparoscopic ureteroneocystostomy psoas hitch and is doing well 3 months postoperatively.Conclusions: This is the first series of laparoscopic ureter reanastomosis reported to date. The outcome was excellent with low postoperative morbidity typically associated with laparoscopy. PMID- 10838382 TI - Long-term urodynamic evaluation of a continent urinary reservoir. AB - Objective: To characterize urodynamic changes among patients who underwent a continent urinary reservoir (Florida Pouch).Methods: Patients who had a continent urinary diversion performed between January 1988 and December 1991 were asked to undergo sequential urodynamic evaluation to compare early (24 mo) changes in reservoir function. The difference in reservoir function was evaluated by defining change in maximum enterocystometric capacity of >/=100 mL, changes in pressure of 5 cm H(2)O, changes in segmental contractions were >/=3. High pressure contractions were those >/=30 mm H(2)O.Results: Seventeen patients underwent sequential urodynamic evaluation. The mean timing of the early study was 12.2 months (range 3-24 mo). The mean timing of the late study was 47.1 months (range 30-58 mo). The mean capacity of the reservoir initially was 698 mL (range 474-1000 mL). On long-term study the mean was 793 mL (range 400-1000 mL). The capacity remained unchanged or increased in 88% of patients. The pressure within the reservoir remained unchanged in seven patients, decreased in eight, and increased in two. Segmental contractions originally occurred in 15 patients with 11 low pressure and 4 high pressure type contractions. On long-term evaluation, 13 patients demonstrated contractions with 10 low pressure and 3 high pressure in character. Of note, 75% of those with initial high pressure contractions had no (1) or low pressure (2) contractions on subsequent evaluation.Conclusion: In long-term urodynamic evaluation of the continent urinary reservoir the reservoir capacity remained unchanged or increased in 88% of patients, while reservoir pressure remained unchanged or decreased in 88%. This long-term evaluation, therefore, demonstrates the continent urinary reservoir to be a low pressure system of adequate capacity thereby minimizing the risk of reflux. PMID- 10838383 TI - Abdominal sacral colpopexy for massive genital prolapse. AB - Objective: A retrospective study to determine the success and complications of abdominal sacral colpopexy in correcting massive genital prolapse over an 8-year period between September 1989 and January 1997.Methods: The charts were reviewed for 3 patients with massive procidentia and 15 patients with symptomatic posthysterectomy vaginal vault prolapse, who desired preservation of sexual function and underwent abdominal sacral colpopexy with Marlex mesh at two community teaching hospitals.Results: In 16 of the 18 patients, one or more concurrent procedures were performed at the same time, including three Burch colpocystourethropexies and one Raz bladder neck suspension, which successfully controlled urinary stress incontinence. In three cases, staging procedures were done for ovarian neoplasia. There were no intraoperative complications. One patient developed a superficial abdominal wound infection, one patient had a deep venous thrombosis in her left leg 7 days postoperatively, and one patient experienced a 1 cm area of graft erosion 10 months postoperative requiring partial resection. Duration of follow-up has varied from 8 months to 5 years. One patient died 43 months after surgery of unrelated causes. No patients developed recurrent prolapse.Conclusions: Abdominal sacral colpopexy is a successful operation for the correction of prolapse. Serious complications are infrequent. Photographs of the technique and a review of the literature are presented. PMID- 10838384 TI - Risk of complications following gynecological laparoscopic surgery. AB - Objective: To determine the incidence and predictors of risk for operative complications, conversions to laparotomy, and postoperative admissions following laparoscopic procedures.Methods: We obtained demographic information and medical history on all 843 women who underwent laparoscopic procedures at the Brigham and Women's Hospital between January and December 1994. All major complications following surgery were recorded. Major operative complications were defined as bowel, bladder, ureter, or vascular injuries, or significant abdominal wall or other internal bleeding. Categorical analysis was used to compare differences in the rates of operative complications, conversions to laparotomy, and postoperative admissions following laparoscopy. We also estimated the influence of specific laparoscopic procedures on the risk of adverse complications following surgery.Results: Operative complications and conversion to laparotomy occurred at rates of 19.0 and 47.4 per 1,000 laparoscopic procedures, respectively. Of 843 women studied, complications included 4 bowel, 2 bladder, 1 ureteral, 2 vascular, and 5 abdominal wall injuries. There were 165 (19.6%) patients admitted postoperatively. Aside from the type of operative procedure, age was the single most important predictor of complications. Women with government or HMO insurance were somewhat more likely to have their laparoscopy converted to a laparotomy than women with private insurance. Relative to all other operative procedures, women receiving laparoscopic-assisted treatment of endometriosis and women undergoing ovarian cystectomy had generally low rates of operative complications, conversions to laparotomy, and postoperative admissions. In contrast, 12.5% of women receiving laparoscopic-assisted vaginal hysterectomy experienced operative injuries or abdominal bleeding and 90.0% were hospitalized postoperatively.Conclusion: Serious operative complications after major gynecologic laparoscopy were rare in this data set. The complexity of the laparoscopic procedure is directly proportional to the rate of operative complications, conversions to laparotomy, and postoperative admissions to the hospital. PMID- 10838385 TI - Risk adjustment for complications of hysterectomy: utility of routinely collected administrative data. AB - Objective: To determine the ability of risk adjustment methods that use routinely collected administrative data to explain variability in complication rates after hysterectomy.Methods: Discharge data on all non-radical hysterectomies performed in North Carolina between January 1988 and September 1994 were obtained from the North Carolina Medical Database Commission. Complications were categorized as 1) surgical (eg, hemorrhage, organ injury) and 2) medical (eg, myocardial infarction, pneumonia). Comorbidities included peritoneal adhesions and chronic medical problems. Hospital charges were adjusted for inflation. Univariate analyses were performed using the Kruskall-Wallis test for skewed continuous variables and chi(2) tests for categorical variables. Multivariate analysis was performed using unconditional logistic regression, with complication rate as the dependent variable.Results: There were 107,648 cases performed at 134 hospitals, with an overall complication rate of 9.5%. When cancer and pregnancy cases were excluded, the surgical complication rate was 5.0%, medical 3.2%. Patients with surgical complications were significantly younger (median age 42 vs 46) and had significantly higher total hospital charges (median $8,127 vs $7,496) than patients with medical complications. Complication rates for individual hospitals varied from 1.5% to 29.3%, with rates highest for academic medical centers (24.3% vs 7.2% for non-teaching hospitals). Significant predictors of complications in univariate analyses included type of hysterectomy, indication, age >/=65 years, insurance status, and teaching hospital status. Coded comorbid conditions were variable in their association with complications. Adjusted odds ratios, controlled for indication and type of procedure, for age >/=65, Medicaid or no insurance, and teaching hospital status were all greater for medical complications than for surgical complications in multivariate analysis (table).The predictive ability of multivariate analysis was better for medical complications than for surgical complications (C-statistic for medical complication model 0.763 vs 0.644 for surgical complications).Conclusions: Surgical complications of hysterectomy are more common, occur in younger women, and are associated with higher hospital charges than medical complications. Complication rates vary widely between hospitals, with teaching hospitals having the highest rates. This difference persists after adjustment for coded comorbidities. Possible explanations for the inability of multivariate analysis to explain the wide range in observed complication rates include 1) shortcomings in the available data or models, such as variability in coding practices between hospitals or variability in surgical difficulty between hospitals that is not captured with ICD-9-CM codes (eg, uterine size, cancer stage) and 2) variability in the quality of care between hospitals. Further research is needed to help determine the role of each of these explanations. Risk adjustment methods that use administrative data based on currently available coding standards are poor predictors of surgical complication rates after hysterectomy and should not be used to reach conclusions about quality of care. PMID- 10838386 TI - Functional outcomes and satisfaction after abdominal hysterectomy. AB - Objective: To compare urinary, lower gastrointestinal (GI), and sexual function and to describe patients' expectations and satisfaction before and after hysterectomy.Methods: Forty-three women completed questionnaires before and about 1 year after abdominal hysterectomy for benign gynecologic conditions. Symptoms related to urinary, lower GI, and sexual function were assessed. Women responded with "agree," "disagree," or "neutral" to statements about treatment options, the decision for hysterectomy, expectations about surgery and recovery, and satisfaction with their relationship with their doctor and their treatment. Comparisons were made between preoperative and postoperative responses. Statistical significance was accepted for P =.002 to account for multiple comparisons.Results: The mean age was 45.4 +/- 6.7 years. Sixty percent were white, and 88% were premenopausal. The most common indication for hysterectomy was myomas in 76%. There were no statistically significant changes in urinary or bowel symptoms before and after hysterectomy with preoperative symptoms resolving in some women after surgery, and developing in others. Fewer women experienced abdominal bloating after hysterectomy than before. Frequency of intercourse and satisfaction with their sexual relationship did not change in the 34 sexually active women. The level of satisfaction with their treatment and their relationship with their doctor was very high.Conclusion: Women experience a high degree of satisfaction with treatment 1 year after abdominal hysterectomy for benign gynecologic conditions. Symptoms related to urinary, GI, or sexual function occur frequently before and after surgery, but hysterectomy does not result in consistent changes. PMID- 10838387 TI - Hysterectomy in a decade of change: more than a weighty matter. AB - Objective: Hysterectomy is the second most common surgical procedure performed in the United States. Interest to decrease use of hysterectomy has focused on developing explicit criteria for surgery, peer review and second opinion, as well as changing reimbursement incentives. The purpose of this study is to describe the change in the number of hysterectomies, the indications for hysterectomies, and the histopathology of hysterectomy specimens over the last 10 years at one private, nonprofit, academic, community hospital that does not have a formal system for monitoring hysterectomy use.Methods: Retrospective study utilizing administrative data matched with pathology reports for nonmalignant hysterectomies performed in 1985, 1990, and 1995. Uterine weight and histopathologic diagnoses were recorded. When pathology reports were unavailable, cases were reviewed to determine if the indication was related to prolapse or urinary incontinence. Outcome measures included number of procedures performed, indications for procedures and uterine pathology (uterine weight, histopathology). We utilized the Cochran-Mantel-Haenszel chi(2) and the Kruskal Wallis rank sum statistic (P <.05).Results: The number of hysterectomies declined from 631 in 1985 to 461 in 1995 (P =.007). The median uterine weight increased (P =.0007), and there was a decrease in the number of specimens with absent histopathology, decreasing from 6% in 1985 to 3% in 1995 (P =.013). This difference was explained by changes in the number of procedures performed on patients with private insurance and HMO reimbursement (P =.018).Conclusions: While the proportion of surgeries performed for specific indications did not change, the number of hysterectomies performed decreased, and there were fewer normal specimens. PMID- 10838388 TI - Prediction of endometrial ablation success by preoperative findings. AB - Objective: To determine the influence of preoperative findings on the outcome of hysteroscopic endometrial ablation.Design: We performed a retrospective chart review of 80 women between the ages of 25 and 50 years who underwent an endometrial ablation for menorrhagia or menometrorrhagia from 1992 to 1996, through a university reproductive endocrinology clinic.Materials and Methods: All eligible patients had a detailed subjective history obtained for duration of dysmenorrhea (Dys) and menorrhagia or menometrorrhagia (Men). Each had a preoperative transvaginal pelvic ultrasound with documentation of the uterine cavity contour and if an intramural myoma was present. A gynecological examination was ascertained from the patients clinical record with regard to uterine size. A benign Papanicolaou smear within 1 year of surgery and a normal endometrial biopsy was required for inclusion. Those with evidence of carcinoma or menopausal symptoms were excluded. Each patient had endometrial preparation with either danazol, GnRHa, or progestin. All ablations were performed by the same surgeon with "rollerball" electrocautery. Those patients who required medical management or additional surgery to control their vaginal bleeding during follow-up were designated as ablation failures. The use of stepwise logistic regression with ablation outcome as the dependent measure was used along with univariate analyses via chi(2) and t test to compare successes and failures on specific characteristics.Results: The sample was divided into two groups, success (group 1) or failure (group 2), and were matched for gravity, parity, and uterine size. Between the two groups there were no significant differences in the duration of menorrhagia/menometrorrhagia or dysmenorrhea. Additional therapy was required in 41% of the study group, designated as failures. The length of follow up was 36 months for group 1 and 27.7 months for group 2. The results are summarized in the table below.The difference in age between the two groups strongly suggests a tendency toward failure with increasing age (P =.06). The diagnosis of a normal uterine cavity preoperatively shows a trend for a successful outcome (P =.07) when compared with the presence of an intracavity lesion, fibroid, or polyp. Those patients with an intramural fibroid had a tendency toward a higher failure rate (P =.10). Comparing the medications used to prepare the endometrium, patients treated with danazol had a trend toward a higher success rate (P =.09) than GnRHa or progestins.Conclusion: Preoperative findings can provide additional information with regard to endometrial ablation success. It appears that the trend toward failure is increased in patients with increased age, the diagnosis of an abnormal uterine cavity by ultrasound, and the presence of an intramural fibroid. Danazol administration, to prepare the endometrium, appears to offer a lower failure rate compared to GnRH agonists or progestins. Patients at greater risk of endometrial ablation failure based on age >/=41 years, abnormal intrauterine cavity, or the presence of intramural fibroids should be counseled about the higher failure rate and consider an alternative procedure such as hysterectomy. PMID- 10838389 TI - Intraoperative blood loss and gestational age at pregnancy termination. AB - Objective: To establish the relationship of measured intraoperative blood loss to gestational age at pregnancy termination, and to determine which factors, if any, affect the risk of bleeding.Methods: A single-operator series of 363 consecutive women undergoing pregnancy termination between 5 and 24 weeks gestational age, as dated by ultrasound, was prospectively evaluated. All pregnancies under 13 weeks gestation were terminated by mechanical dilation and suction curettage without preoperative cervical ripening. All pregnancies between 13 and 24 weeks gestation were terminated by preoperative osmotic cervical dilation with laminaria tents and subsequent uterine evacuation by a combination of suction curettage, sharp curettage, and Bierer forceps extraction. All patients over 12 weeks gestation received a postoperative oxytocin infusion. Whenever possible, amniotic fluid and blood were collected and measured separately. Patients were excluded from the data analysis for pregnancy demise, PPROM, Potter's syndrome, or inability to separate blood establish their relationship. After adjustment for gestational age, the results were analyzed to determine if blood loss was related to maternal age, smoking history, body habitus, or operative indication.Results: A curvilinear relationship between blood loss and gestational age was observed. Mean blood loss at 24 weeks exceeded 800 mL. After adjustment for gestational age, no factors significantly affected blood loss at dilation and aspiration of first trimester pregnancies. In those patients undergoing dilation and evacuation in the second trimester, both simple and stepwise regression analyses showed obesity (BMI >/=32.3) to be significantly associated with increased blood loss (P <.05). Neither age, parity, previous cesarean section, nor smoking history were significantly associated with increased blood loss at dilation and evacuation.Conclusions: With advancing gestational age, intraoperative blood loss increases in curvilinear fashion. Termination providers should be advised that, although blood loss is unaffected by many factors, obese patients are at risk for increased bleeding at dilation and evacuation of pregnancies beyond 12 weeks gestation. PMID- 10838390 TI - Clinical outcomes of OPERA, out-patient endometrial resection/ablation. AB - Objective: To evaluate the clinical outcomes of 18-month follow-up for OPERA, Out Patient Endometrial Resection/Ablation using the (OPERA)Star tissue aspirating resectoscope.Methods: Patients suffering from abnormal uterine bleeding (AUB) and/or submucosal fibroids who where indicated for surgical intervention underwent OPERA for treatment of their condition using the (OPERA)Star tissue aspirating resectoscope. This resectoscope contains a tissue aspiration and morcellation channel for continuous removal of tissue strips. Follow-up assessments were conducted at 1 month, 3 months, 6 months, 12 months, and 18 months to evaluate long-term clinical results of the procedure. Data collected include current level of bleeding, subsequent procedures to manage condition, and patient's level of satisfaction.Results: A total of 33 patients presented with AUB requiring surgical intervention for relief of their symptoms. Uterine cavity averaged 10.7 cm (+/- 2.2), range 6.7-13.4 cm). Complete resection and ablation of the endometrium and submucosal fibroid(s) if present, was performed. Current 18-month follow-up data indicate that 21 (68%) patients achieved amenorrhea and 8 (26%) patients achieved hypomenorrhea. Two patients were lost to follow-up. Two patients underwent a subsequent procedure; one repeated OPERA and one hysterectomy for fibroids. Patient's overall satisfaction rate with the results of their procedure was 97%.Conclusion: OPERA is an effective minimally invasive method for management of abnormal uterine bleeding. At 18 months, clinical outcomes of OPERA indicate an optimal level of patient satisfaction for management of their bleeding with 94% of patients achieving either amenorrhea or hypomenorrhea. Continuous aspiration and morcellation during resection keeps the operative field clear of debris and simplifies the procedure. PMID- 10838391 TI - Clinical experience with the (OPERA)StarSL using saline irrigation fluid for OPERA. AB - Objective: To present clinical results using a new hysteroscopic resectoscope, the (OPERA)Star(SL) with PEARL technology, which cuts and coagulates uterine tissue in a physiologic, conductive fluid environment in order to perform OPERA, Out-Patient Endometrial Resection/Ablation. Use of physiologic normal saline in resectoscopy has not previously been feasible. Use of normal saline prevents complications such as hyponatremia and cerebral edema due to excessive absorption of traditional nonphysiologic fluids.Methods: Patients suffering from abnormal uterine bleeding with or without submucosal fibroids who were scheduled for surgical intervention underwent OPERA for treatment of their condition. The (OPERA)Star(SL) resectoscope with PEARL technology was used with a 0.9% saline distention medium. The PEARL technology insulates the active electrode so that a noninsulating fluid, such as 0.9% saline or lactated ringers, can be used while maintaining the ability to cut and coagulate tissue using a standard monopolar electrosurgery unit.Operative data recorded include uterine size, presence and size of fibroids, pretreatment with endometrial thinning agents, operative time, cutting/coagulating power levels, estimated blood loss, and calculated fluid deficit. Additional data include the ability to cut and coagulate tissue as compared to standard resectoscopes used in the physician's clinical practice.Results: A total of 22 patients underwent operative hysteroscopy with the described resectoscope. Average age of all patients was 45 years (range 30 70), and average weight was 158 pounds (range 106-258). Eighteen (82%) patients underwent OPERA while the remaining 4 (18%) patients underwent fibroid resections. The average uterine size was 7 weeks (range 4-14). Average operative time was 26 minutes, and all procedures were successfully completed without complications. Fluid deficit averaged 480 mL (range 0.0-1.54 L) as reported by the Flo-Stat Fluid Management System.A Likert scale was used to rate the device's ability to cut and coagulate tissue with results indicating the (OPERA)Star(SL) accomplishes the same clinical results to standard resectoscopic devices used in the physician's clinical practice.Conclusion: Complete uterine tissue resection and ablation can now be accomplished in a saline environment. The unique PEARL technology with the (OPERA)Star(SL) resectoscope is effective in cutting and coagulating uterine tissue. Operating in a saline environment eliminates the most prevalent potential complication of operative hysteroscopy when using nonphysiologic irrigation solutions, dilutional hyponatremia, and secondary cerebral edema. PMID- 10838392 TI - The BirthPlace collaborative practice model: results from the San Diego Birth Center Study. AB - Objective: The search for quality, cost-effective health care programs in the United States is now a major focus in the era of health care reform. New programs need to be evaluated as alternatives are developed in the health care system. The BirthPlace program provides comprehensive perinatal services with certified nurse midwives and obstetricians working together in an integrated collaborative practice serving a primarily low-income population. Low-risk women are delivered by nurse-midwives in a freestanding birth center (The BirthPlace), which is one component of a larger integrated health network. All others are delivered by team obstetricians at the affiliated tertiary hospital. Wellness, preventive measures, early intervention, and family involvement are emphasized. The San Diego Birth Center Study is a 4-year research project funded by the U.S. Federal Agency for Health Care Policy and Research (#R01-HS07161) to evaluate this program. The National Birth Center Study (NEJM, 1989; 321(26): 1801-11) described the advantages and safety of freestanding birth centers. However, a prospective cohort study with a concurrent comparison group of comparable risk had not been conducted on a collaborative practice-freestanding birth center model to address questions of safety, cost, and patient satisfaction.Methods: The specific aims of this study are to compare this collaborative practice model to the traditional model of perinatal health care (physician providers and hospital delivery). A prospective cohort study comparing these two health care models was conducted with a final expected sample size of approximately 2,000 birth center and 1,350 traditional care subjects. Women were recruited from both the birth center and traditional care programs (private physicians offices and hospital based clinics) at the beginning of prenatal care and followed through the end of the perinatal period. Prenatal, intrapartum, postpartum and infant morbidity and mortality are being compared along with cost-effectiveness and acceptance of the model by patients. Data collection occurred primarily through medical record abstraction with the addition of two patient questionnaires. Comparability of the cohorts was established by using a validated methodology to determine medical/perinatal risk and birth center eligibility, which included assessment by two CNMs and an independent blind review by a perinatologist. The cost analysis uses a resource utilization approach and new methodologies such as activity-based-costing to compare costs from both the perspective of the payor and the health care provider. Patient satisfaction was measured using a self-administered patient questionnaire.Results: Current preliminary results from approximately 38% of the final expected study sample are available. Crude and adjusted analysis have been conducted. Overall, the preliminary results suggest similar morbidity and mortality in the two groups. Fetal deaths are 0.75% in the index and 0.64% in the comparison group, with early neonatal deaths at 0.26% and 0.23%, respectively. The traditional care group showed adjusted rate differences of 5.83% more major maternal intrapartum complications and 9% more NICU admissions. While the birth center group showed adjusted rate differences of 5.5% more low birth weight and 0.95% more preterm birth. For other outcomes, the birth center group showed an adjusted rate difference of 22.34% more exclusive breastfeeding at discharge. Also, there was less utilization of cesarean section and assisted delivery in the birth center group as compared to the traditional care group. The adjusted rate difference for normal spontaneous vaginal deliveries in nulliparas was 10.23% more in the birth center group, with similar results in multiparas with and without history of cesarean (28.88% and 7.84%, respectively). Preliminary results also show that the average total cost for pregnancy-related services paid by California Medicaid was $4,550 for the birth center and $5,535 for the traditional care group. Final results based on the full study sample (full data available February 1998) details of payor costs such as provider, facility, NICU, and ancillary along with costs from the health care system perspective and patient satisfaction results will be presented.Conclusion: Current results suggest similar morbidity and mortality between the birth center program and traditional care groups, with less resource utilization translating to lower costs in the collaborative practice model. Results suggest that collaborative practice using a freestanding birth center as an adjunct to an integrated perinatal health care system may provide a quality, lower-cost alternative for the provision of perinatal services. PMID- 10838393 TI - The use of midwives as first assistants in cesarean section. AB - Introduction: In 1997, medical insurance gives limited financial reimbursement to physicians who first assist cesarean sections. Therefore, a formal program was developed at our community teaching hospital using midwives in place of physicians as first assistant in cesarean section. Midwives on the midwifery service were taught the principles and procedures of obstetrical anesthesia, anatomy, and physiology of the gravid abdomen.A team approach using obstetrician/gynecologist, anesthesiologist, midwives, and operating nurses performed the tracking in a 1-day course. A 28-minute video was developed to depict the role of the midwife as first assistant. Additionally, the principles of operation room set-up and instrumentation were taught.Result: All 24 midwives on the midwifery service were involved in the teaching of first assisting an attending obstetrician. After taking the course, nurse-midwives received formal certification from the department of obstetrics and gynecology and were approved by risk management. They also received clinical privileges outlining these guidelines. Their malpractice rates have not increased. The time to complete a cesarean section has not increased for the physician, and an emergency cesarean section is no longer delayed by waiting for an assistant to arrive. Patient satisfaction has increased because of decreased waiting time for a cesarean section and increased familiarity with the entire operating team.Discussion: Midwives can be trained to first assist the obstetrician/gynecologist in a cesarean section. Since the duration of a cesarean section is not prolonged, the patient will not experience increased blood loss or infection secondary to a prolonged procedure. The patient is more relaxed since they know that their provider during labor is also one of their surgeons for the cesarean section. PMID- 10838394 TI - The impact of prenatal care coordination on birth outcome. AB - Objective: Care coordination is a Medicaid benefit program designed to assist low income women in gaining access to pregnancy related services. This study examines the relationship between prenatal care coordination and its impact on access to prenatal services and birth outcome.Methods: Ninety-four women who received prenatal care coordination and gave birth at Wishard Memorial Hospital in 1996 comprised the study population. A control group of 190 non care coordination participants was obtained from women who gave birth immediately before and after the women who was a care coordination participant. The prenatal and birth records from both groups were retrospectively reviewed. Data collected included information on birth weight, gestational age at delivery, WIC participation, number of prenatal visits, smoking and substance abuse. Perinatal complications were also recorded. Statistics was by t test and chi(2) with significance at.05.Results: Women enrolled in care coordination received 10 prenatal visits versus 8 for controls (P =.0005). Preterm labor occurred in 1.1% of women in care coordination compared to 5.8% of controls (P =.063). There was no difference in birth weight or the incidence of birth weight <2500 g between care coordination and controls. WIC participation was greater in the care coordination group, 95.7% versus 78% for controls, P <.05. In addition, women with care coordination were less likely to smoke during pregnancy.Conclusions: Care coordination does improve access to prenatal services as evidenced by an increase in prenatal visits and enrollment in WIC services. However, this study failed to demonstrate a direct relationship between care coordination and improved birth outcome. PMID- 10838395 TI - Cost consequences of implementation of an early obstetrical discharge program in a military teaching hospital. AB - Objective: To evaluate the cost consequence of an early obstetrical discharge program in a military teaching hospital.Methods: The study involved a control group of routine obstetrical discharge patients with uncomplicated vaginal delivery from March to August 1994 and the study group of early obstetrical discharge (24-48 hours) patients with uncomplicated vaginal delivery from March to August 1996.Results: There were 1,042 total control patients with routine vaginal delivery from March to August 1994 totaling 2,668 hospital days with a mean number of hospital days of 2.56 per patient. The study group of early obstetrical discharge patients from March to August 1996 with uncomplicated vaginal delivery encompassed 1,050 patients with 1,965 days with mean hospital days of 1.87 per patient. The total cost of admissions (cost calculation of $1,221/hospital day) fell from $3,257,628 in the routine discharge group to $2,399,625 in the early discharge cohort showing a total cost savings of $858,003 over the 6-month study period. The average cost per admission fell from $3,126/day to $2,285/day without an increase in the postpartum pediatric or maternal adverse outcomes.Conclusion: An early obstetrical discharge program at a military teaching hospital showed significant cost savings without concomitant increase in pediatric or maternal adverse outcomes. PMID- 10838396 TI - Applying no-fault compensation criteria to obstetric malpractice claims. AB - Objective: To determine in obstetric malpractice cases the incidence of adverse events (AE), defined in the litigation literature as an injury caused by medical management, and to determine the proportion of cases that would meet no-fault criteria.Methods: In a retrospective cohort design, all neonatal outcome claims (n = 47) occurring between January 1992 and December 1993 were reviewed from carriers in Massachusetts, Colorado, and Utah. Two obstetricians determined whether an AE occurred, as defined above. The cases were then reevaluated according to Swedish Compensatory Event (SCE) criteria, which compensate AEs due to avoidable or substandard care, and to Florida Neurological Injury Compensation Association (NICA) criteria, which compensate neurologic injury following peripartum oxygen deprivation or mechanical injury.Results: An AE occurred in only 23/47 claims, representing 49% (95% CI 34-64%). SCE criteria were met in all 23 of the cases with an AE. NICA criteria were met in 9/23, or 39% (95% CI 19 61%), of the cases with an AE. Neither SCE nor NICA criteria were satisfied in any of the 24 cases without an AE. There were strong relationships between an occurrence of an adverse event and fulfilling criteria for SCE (P <.001) and NICA (P <.001).Conclusion: Neonatal injury caused by medical management occurs in less than half of the obstetric malpractice claims. No-fault compensation criteria can be successfully applied to claims, and both the SCE and NICA methods are specific in avoiding compensation to claims without an AE. The SCE criteria are more sensitive than the NICA criteria for compensating AEs. Applying the SCE system to claims would result in a larger proportion of claims successfully meeting no fault criteria than using the NICA system. PMID- 10838397 TI - The evaluation of the health care needs of women with disabilities. AB - There are 27 million disabled women in the United States. These women face many unique challenges in accessing medical care and frequently report an insensitivity to their specific health care needs. Disabled women also have a wide array of health-related issues, including multiple concurrent medical disorders, psychosocial issues, and socioeconomic concerns. Their overall experience is that of a fragmented health care delivery system that often is inadequate in many important areas such as reproductive health, post-reproductive health, sexuality, etc.In order to address these issues, the departments of neurology and gyn/ob formed a multidisciplinary Women's Health Collaborative Research Group (WHCRG). Its mission is to develop a wellness model of comprehensive women-centered care that meets the special health care requirements of women with chronic disabling conditions. The Women's Health Issues Survey is a 72-item self-report needs assessment that was administered to over 300 women with multiple sclerosis, stroke, and other neurologic disorders. The results of the survey were critical to the planning and implementation of a center for women with disability.Approximately 50% of the women (median age 45 years) were moderately to severely disabled, as measured by the Kurtzke Expanded Disability Status Scale. Over 20% of those surveyed reported a history of uterine fibroids, endometriosis or ovarian cysts, irregular menstrual periods, and deterioration of their neurologic condition (eg, multiple sclerosis) during menstruation. Forty percent were sexually inactive. Seventy percent of the postmenopausal cohort were not taking hormone replacement therapy.The ability to identify and survey a representative sample of disabled women and develop a database linking demographic, physical disability, economic, and health care access information is a key step in implementing a cross-functional approach toward improving the level and scope of health care for disabled women. PMID- 10838398 TI - A multicenter randomized trial of DIAC for cyclical mastopathy. AB - Cyclical mastopathy (CM) (ie, fibrocystic disease) is a commonly occurring condition with severe breast pain reportedly affecting up to 70% of women. DIAC is a new, non-hormonal treatment for CM that is administered as a capsule taken orally once each day. Earlier trials of DIAC have yielded promising results with resolution of the signs and symptoms of CM. We report the largest randomized clinical trial published to date on therapy for CM.Objective: To evaluate the safety and efficacy of different doses of DIAC compared to placebo given for 6 months to women with CM.Methods: A double-blind, randomized, dose-response, parallel-group clinical trial design was used. All enrolled women had moderate or severe cyclical mastalgia for at least 12 of the prior 24 months associated with nodularity and/or fibrosis on clinical exam. After obtaining informed consent, women were examined to exclude breast cancer or suspicious breast masses, thyroid disease, or other cancers. No one had recent breast surgery nor change in the use of hormones or adjuvant medication for breast pain for at least 3 months. A randomly generated number sequence was used to create a block randomization scheme. Women were assigned to receive either one of 5 daily doses of DIAC or placebo. One of the doses of DIAC was expected to be too low to provide relief of mastalgia. Women kept diary data daily. Clinical examinations and quality-of-life questionnaires were completed during the luteal phase of the menstrual cycle before and after 1, 2, 4, and 6 months of study therapy. Women were contacted by phone during the luteal phase in months 3 and 5 for additional data retrieval.Endpoints of the study included breast examination graded by quadrant for tenderness, size/amount of nodularity, and fibrosis. These data were converted to a standardized score for tenderness (TS) and nodularity (NS). Daily pain assessment from the diary was converted to a standardized pain score (PS). A total CM score represented the sum of these three measures. Changes in quality of life during and after DIAC therapy were also evaluated. The study had the power to detect at least a 30% improvement in the total CM score with beta = 0.80 and alpha = 0.05.Results: Twenty-six sites in 16 states within the United States randomized a total of 385 women with CM. The mean age of women enrolled was 42 years (range 21-73). Over two-thirds of the women had completed study drug intervention as of September 1997. All study drug therapy was to be completed by December 1997 with unblinding in January 1998. No serious or life-threatening side effects had been observed in the trial as of September 1997.Conclusions: DIAC is a promising new therapeutic for CM. The unblinded data from the completed trial assessing safety, efficacy, and quality of life will be presented. PMID- 10838399 TI - The Place of Whole-Body PET FDG for the Diagnosis of Distant Recurrence of Breast Cancer. AB - Purpose: To study the role of positron emission tomography 18F-fluorodeoxyglucose (PET FDG) imaging in patients with a suspicion of breast cancer recurrence.Procedures: Whole-body PET FDG was performed in 39 women. Thirty-four were included because of asymptomatic tumor marker increase. PET findings were confirmed by oriented imaging or by biopsy. Follow-up data were collected over a period of at least 12 months.Results: PET FDG depicted 37/39 sites in 31/33 patients with recurrence. PET missed one locoregional recurrence and in one patient peritoneal carcinomatosis developed 6 months after a negative PET. False positive PET FDG corresponded to lung infection, degenerative bone disease, and reconstruction artifact. The conventional imaging work-up depicted sites of recurrence in 6/33 patients.Conclusion: Whole-body PET FDG is highly sensitive for the detection of distant breast cancer recurrence. Prospective studies are mandatory to address its potential impact on patient management and survival. PMID- 10838400 TI - Evaluation of the Effect of Radiation Therapy to Nasopharyngeal Carcinoma by Positron Emission Tomography with 2- AB - Purpose: This investigation evaluated the effectiveness of positron emission tomography with 2-[F-18]fluoro-2-deoxy-D-glucose (PET-FDG) in assessing residual tumor or tumor recurrence in postradiation nasopharyngeal carcinoma (NPC) patients.Procedures: Forty-six patients with histologically proven NPC who received radiotherapy were included. PET-FDG images were analyzed by a semiquantitative method, metabolic ratio (tumor to cerebellum ratio).Results: The overall sensitivity and specificity of PET-FDG to detect residual tumors and recurrent lesions in the postradiation patients were 80% (12/15) and 87% (27/31), respectively. In patients with PET-FDG 6 months after radiation therapy, the sensitivity and specificity raised to 92% (11/12) and 100% (20/20), respectively.Conclusions: PET-FDG is effective in the evaluation of NPC treated with radiation. The optimal timing in assessing residual tumor or tumor recurrence in postradiation patients should be 6 months or later. PMID- 10838401 TI - Tumor Burden Assessment with Positron Emission Tomography with AB - Objective: In patients with advanced cancer, total tumor burden affects the likelihood of tumor response and has important implications for prognosis. The aim of this study was to select the optimum 2-[F-18]fluoro-2-deoxy-D-glucose positron emission tomography (FDG PET) tumor uptake parameter to accurately measure tumor burden in advanced metastatic renal cell cancer, in comparison with volumes measured with computed tomography (CT), as a reference test.Materials and Methods: Six patients with metastatic renal cell carcinoma measurable on CT were studied. CT and FDG PET scans were carried out on all patients within 4 weeks prior to their entry into a phase I-II radioimmunotherapy trial. CT-based evaluation of disease extent (tumor volume) and 4 PET-based measurements (standardized uptake value[SUVmax], SUVav, volume, and total lesion glycolysis [TLG]) were performed independently by a radiologist (VN) and a nuclear medicine physician (TA). The degree of correlation between conventional (CT) extent of disease and parameters describing tumor concentration of FDG was then determined.Results: Fifty-seven CT-measurable metastatic lesions in lung, abdomen, and scalp were evaluated in 6 patients. There was a high correlation between CT and FDG PET volume estimates for lesions greater than 5 cm(3) in size. However, a PET-derived parameter that embodies both FDG uptake and lesion size, the TLG, correlated better with CT-derived tumor volume than did FDG PET volume alone.Conclusion: Using CT volume as a gold standard, the optimal PET-based estimate of total tumor burden in patients with metastatic renal cancer is the sum over all lesions of the total lesion glycolysis. PMID- 10838402 TI - The Status of Positron Emission Tomography in China. PMID- 10838403 TI - F-18 FDG Accumulation in an Octreotide Negative Merkel Cell Tumor. AB - Regional positron emission tomography (PET) imaging with F-18 Fluorodeoxyglucose (FDG) was performed in a patient with pathologically proven Merkel cell tumor around the right knee region. Prior to the PET imaging, whole-body Indium-111 octreotide scan was performed in this patient but was negative. F-18 FDG was offered as an attempt to image this somatostatin-receptor negative Merkel cell tumor. The PET images delineate a series of focal abnormal uptake along the right lower extremity without evidence of distant metastasis. Patient was treated locally. The positive accumulation of F-18 FDG in Merkel cells may offer a tool for defining the extent of this rare neuroendocrine tumor. PMID- 10838404 TI - Temporal Lobe Hypometabolism Ipsilateral to a Hypothalamic Mass. Relationship to Gelastic Seizures. AB - The purpose of this study was to investigate metabolic changes associated with a right hypothalamic mass in a 26-year-old gelastic seizure patient. Positron emission tomography (PET) imaging of the brain was performed in the interictal state using 18F-fluorodeoxyglucose (18F-FDG) in this patient. Temporal lobe hypometabolism was noted ipsilateral to the hypothalamic lesion. The mass itself had little to no uptake of 18F-FDG. This is the first known PET imaging report of temporal lobe hypometabolism ipsilateral to a presumed hypothalamic hamartoma causing gelastic seizures. Further studies are needed in other patients to test whether interictal PET imaging may help plan the removal of epileptogenic hypothalamic lesions. PMID- 10838405 TI - Response of Osteosarcoma to Chemotherapy. Evaluation with F-18 FDG-PET Scans. AB - Objective: Positron emission tomography (PET) using fluorine-18-fluoro-2-D deoxyglucose (FDG) is increasingly being used to evaluate and manage oncology patients. Several reports have documented its utility in diagnosis, staging, response to treatment, and tumor viability assessment. There is, however, a paucity of literature on PET scanning in patients with osteosarcoma. We report results of serial F-18 FDG-PET scans in 16 untreated patients with osteosarcoma who underwent chemotherapy prior to surgical resection of the primary tumor site.Procedure: Changes in tumor fluoro-2-D-deoxyglucose (FDG) uptake were correlated with percent tumor necrosis on histopathology. PET studies were analyzed by visual assessment of tumor uptake of FDG by 3 independent observers, calculating a tumor to normal background activity ratio (TBR) by drawing regions of interest (ROIs) around the tumor and background activity in the contralateral normal limb, and percent change in TBR values between baseline and presurgical study.Results: All patients had positive baseline scans. Baseline TBRs ranged between 2.5-8.7 and visual assessment of intensity of FDG uptake was 2-3 on a scale of 0-3. At histopathologic examination, 8 patients were classified as good responses with more than 90% tumor necrosis and 8 patients as poor responses with less than 90% necrosis. Tumor necrosis was accurately predicted on PET scan in 15/16 patients by visual assessment, 14/15 patients by final TBR value on presurgery scans, and 7/15 patients using percent change of TBR on serial scans.Conclusions: The results of this small series suggest that FDG-PET scanning is fairly accurate in evaluating the response of osteosarcoma to chemotherapy. Visual assessment and TBR are more accurate in predicting tumor necrosis than percent change in TBR on serial scans. PMID- 10838406 TI - High Throughput Scintillation Proximity Assay for the Identification of FKBP-12 Ligands. AB - A high throughput scintillation proximity assay (SPA) was developed to identify novel ligands of FKBP-12, an immunophilin with peptidyl prolyl isomerase (rotamase) activity. Recombinant histidine-tagged FKBP-12 was expressed in Escherichia coli, purified by metal ion affinity chromatography, and immobilized to SPA beads by an antibody that recognizes the histidine tag of the recombinant protein. Using 1 nM [3H] FK506, a well-known macrolid ligand of FKBP-12, specific binding was saturable and accounted for 95% of total binding. Analysis of saturation and homologous displacement isotherms indicated the existence of a single binding site with a Kd value of 1.6 nM. The specificity of [3H] FK506 binding was demonstrated in displacement experiments and showed that rapamycin, another macrolid, was as active as FK506 (IC50 of 3.5 and 3.2 nM, respectively), whereas GPI-1046, a prototype of small molecular compounds with neurotrophic properties and affinity for FKBP-type immunophilins, was more than 1000-fold less active. The high signal-to-noise ratio of 30, together with small standard deviations, makes this novel assay well suited for automated high throughput screening. PMID- 10838407 TI - A Melanophore-Based Screening Assay for Erythropoietin Receptors. AB - A rapid, functional assay in frog melanophore cells for the erythropoietin receptor (EPOR), a member of the cytokine receptor family, is demonstrated. A chimeric receptor that comprised the extracellular portion of the murine EPOR and the transmembrane and intracellular domains of the human epidermal growth factor receptor (EGFR) was subcloned into the expression vector pJG3.6. When the full length EGFR was expressed in melanophores, EGF but not EPO mediated pigment dispersion in a time- and dose-dependent manner with an EC50 of 12.6 6 2.9 pM. However, when the chimeric EPOR/EGFR was expressed, EPO but not EGF stimulated pigment dispersion in a time- and dose-dependent manner with an EC50 of 380 6 107 pM. Neither EGF nor EPO had any effect on pigment dispersion in wild-type melanophores. EGF- and EPO-mediated pigment dispersion was blocked by the bis indolylmaleimide protein kinase C inhibitor Ro 31-8220. This study extends the use of the melanophore-based bioassay to include cytokine receptors in addition to G protein- and tyrosine kinase-coupled receptors. It represents a potentially powerful method for screening of combinatorial libraries to identify novel small molecule agonists and antagonists to this clinically important class of binding sites as well as performing studies of functional ligand-receptor interactions. PMID- 10838408 TI - Natural Products in High Throughput Screening: Automated High-Quality Sample Preparation. AB - At present, compound libraries from combinatorial chemistry are the major source for high throughput screening (HTS) programs in drug discovery. On the other hand, nature has been proven to be an outstanding source for new and innovative drugs. Secondary metabolites from plants, animals, and microorganisms show a striking structural diversity that supplements chemically synthesized compounds or libraries in drug discovery programs. Unfortunately, extracts from natural sources are usually complex mixtures of compounds, often generated in time consuming and, for the most part, manual processes. Because quality and quantity of the provided samples play a pivotal role in the success of HTS programs, this poses serious problems. In order to make samples of natural origin competitive with synthetic compound libraries, we devised a novel, automated sample preparation procedure based on solid-phase extraction (SPE). By making use of modified Zymark (Hopkinton, MA) RapidTrace(r) SPE workstations, we developed an easy-to-handle and effective fractionation method that generates high-quality samples from natural origin, fulfilling the requirements for an integration in high throughput drug discovery programs. PMID- 10838409 TI - High Throughput Quantitation of cAMP Production Mediated by Activation of Seven Transmembrane Domain Receptors. AB - Impairment of G protein-coupled seven-transmembrane (7 TM) receptor function has been implicated in a variety of different pathologic conditions, suggesting that the discovery of specific antagonists may lead to the development of successful therapeutic agents. The effect of different agents on receptor-ligand interaction is often measured directly in a receptor binding assay; however, this assay format can be time consuming and does not detect agents that interact at sites distal to the native ligand binding site. Cyclic adenosine monophospate (cAMP) represents a ubiquitous second messenger generated in response to ligand binding to many 7 TM receptors. The present study describes the practical adaptation of scintillation proximity methodology, using FlashPlate (NEN Life Sciences, Boston, MA) technology to evaluate cAMP production. The bioassay is based on competition between endogenously produced cAMP and exogenously added radiolabeled [125I] cAMP. Cyclic AMP capture is mediated through an anti-cAMP antibody onto a microplate well surface. Removal of unbound radioligand is not necessary because only ligand within #20 mm of the plate surface is detected due to the proximity effect. The data indicate that the use of scintillation proximity technology allows accurate and specific evaluation of G protein-coupled receptor function as measured by cAMP production and is suitable for high throughput screening. PMID- 10838410 TI - Development of an Immunofluorometric, High-Capacity, Cell-Based Assay for the Measurement of Human Type I and Type II Interferons. AB - We have developed a cell-based 96-well microtiter plate, high throughput assay for measuring both type I and type II interferon (IFN) activity on human cells. This assay makes use of a previously described IFN-specific reporter stably expressed in human HT 1080 cells. The induction of the reporter by IFN is determined by measuring the IFN-dependent expression of CD2 on the cell surface. The cytokine-induced expression of CD2 occurs within 48 h and is measured using a time-resolved fluorometric immunoassay. The limit of detection for type I IFN is.0.4 IU/ml. Interassay and intraassay coefficients of variation were 1.1% and 1.3% for the medium control (31 IU IFNb1b/ml), respectively. The limit of detection for type II IFN is.8 IU/ml, and the assay coefficients of variation are similar to those determined for type I IFNs. The level of sensitivity for this assay is comparable to other assays commonly used to measure IFN activity on cells. The current assay has an advantage over antiviral and antiproliferative assays, in that there is no requirement for the use of pathogenic virus or for determining viable cell numbers. The current assay is ideally suited for increasing sample screening and high-capacity automation, making it an excellent tool for drug discovery. PMID- 10838411 TI - Automation Basics: Robotics vs. Workstations. PMID- 10838412 TI - Allegrotrade mark: Moving the Bar Upwards. PMID- 10838413 TI - Agrochemical Research and Development in the 21(st) Century: Bloom or Bust. AB - Economic, environmental, and political issues arising in the last two decades have changed agricultural practice worldwide. While the agrochemical market has diminished, time and cost of R & D and regulatory restrictions have increased, thereby reducing profit margins dramatically. This has led to a change of strategy from screening whole organisms to screening diverse compounds, with high throughput screening processes similar to those used in drug discovery. For large scale high-throughput screening to efficiently facilitate storage, retrieval, preparation and tracking of compounds, the sample management process needs to be highly automated. The introduction of modern automated technology will help to address the problems faced by the agrochemical industry. PMID- 10838414 TI - A Simple Statistical Parameter for Use in Evaluation and Validation of High Throughput Screening Assays. AB - The ability to identify active compounds (3hits2) from large chemical libraries accurately and rapidly has been the ultimate goal in developing high-throughput screening (HTS) assays. The ability to identify hits from a particular HTS assay depends largely on the suitability or quality of the assay used in the screening. The criteria or parameters for evaluating the 3suitability2 of an HTS assay for hit identification are not well defined and hence it still remains difficult to compare the quality of assays directly. In this report, a screening window coefficient, called 3Z-factor,2 is defined. This coefficient is reflective of both the assay signal dynamic range and the data variation associated with the signal measurements, and therefore is suitable for assay quality assessment. The Z-factor is a dimensionless, simple statistical characteristic for each HTS assay. The Z-factor provides a useful tool for comparison and evaluation of the quality of assays, and can be utilized in assay optimization and validation. PMID- 10838415 TI - Quantification of G-Protein Coupled Receptor Internatilization Using G-Protein Coupled Receptor-Green Fluorescent Protein Conjugates with the ArrayScantrade mark High-Content Screening System. AB - Many G-protein coupled receptors (GPCRs) undergo ligand-dependent homologous desensitization and internalization. Desensitization, defined as a decrease in the responsiveness to ligand, is accompanied by receptor aggregation on the cell surface and internalization via clathrin-coated pits to an intracellular endosomal compartment. In this study, we have taken advantage of the trafficking properties of GPCRs to develop a useful screening method for the identification of receptor mimetics. A series of studies were undertaken to evaluate the expression, functionality, and ligand-dependent trafficking of GPCR-green fluorescent protein (GFP) fusion conjugates stably transfected into HEK 293 cells. These GPCR-GFP expressing cells were then utilized in the validation of the ArrayScantrade mark (Cellomicstrade mark, Pittsburgh, PA), a microtiter plate imaging system that permits cellular and subcellular quantitation of fluorescence in whole cells. These studies demonstrated our ability to measure the internalization of a parathyroid hormone (PTH) receptor-GFP conjugate after ligand treatment by spatially resolving internalized receptors. Internalization was time- and dose-dependent and appeared to be selective for PTH. Similar results were obtained for a beta(2)-adrenergic receptor (beta(2) AR)-GFP conjugate stably expressed in HEK 293 cells. The internalized GFP-labeled receptors were visualized as numerous punctate 3spots2 within the cell interior. An algorithm has been developed that identifies and collects information about these spots, allowing quantification of the internalization process. Variables such as the receptor-GFP expression level, plating density, cell number per field, number of fields scanned per well, spot size, and spot intensity were evaluated during the development of this assay. The method represents a valuable tool to screen for receptor mimetics and antagonists of receptor internalization in whole cells rapidly. PMID- 10838416 TI - A Fluorescence-Based High Throughput Screen for the Transporter Associated with Antigen Processing. AB - The transporter associated with antigen processing (TAP) is essential for antigen presentation by major histocompatibility complex (MHC) class I molecules. Traditional methods used to analyze peptide transport mediated by TAP require radioactive labeling of peptides and time-consuming manipulation of Concanavalin A-Sepharose. Drug discovery research requires rapid and reliable evaluation of large number of samples for bioactivity. To meet these requirements a nonradioactive, HTS assay for peptide transport activity of TAP has been developed. The radioactive label in the traditional assays has been replaced by a fluorescent label without compromising the transport efficiency of labeled peptide or the sensitivity of the assay. The use of multiscreen filtration plates has facilitated higher throughput and eliminated the centrifugation steps used in traditional TAP assays. The HTS assay shows similar kinetic characteristics as compared to the traditional assay. The HTS assay has been adapted on a Quadratrade mark 96-320 96-channel pipetting station (Tomtec, Hamden, CT) by optimizing time course, dose response of TAP to peptides and adenosine triphosphate (ATP), signal/noise ratio, reproducibility, and reagent stability. This HTS system has been utilized to screen a multiplexed compound library with a maximum of throughput 17,600 compounds per week. PMID- 10838417 TI - A Target-Specific Cellular Assay for Screening of Topoisomerase I Inhibitors. AB - We have developed a cellular, target-specific high-throughput assay for the detection of topoisomerase I inhibitors. Topoisomerase I is a nonessential enzyme involved in controlling DNA topology. Topoisomerase I is the target of anticancer drugs such as camptothecin as well as a candidate target for new antifungal drugs. A wild-type Saccharomyces cerevisiae strain and its isogenic topoisomerase I deletion mutant (DeltatopI) were labeled with S65T and wild-type green fluorescent protein (GFP), respectively. We showed that the growth of such a pair of S. cerevisiae strains labeled with this GFP combination can be independently quantified after both strains were mixed. When growth of the mixture of wild-type and DeltatopI strain was monitored in the presence of compounds, only growth of the wild-type strain was inhibited by the topoisomerase I-specific drug camptothecin. In contrast, amphotericin B, a broad-spectrum antifungal drug, inhibited growth of both strains. The two strains were used to screen compound libraries. While 0.9% of all compounds inhibited growth of both strains, only 0.06% inhibited the wild-type but not the DeltatopI strain. Thus, by using a DeltatopI strain as internal control in the same assay mixture, the number of candidate topoisomerase I inhibitors to be retested could be reduced by more than 90%. Further applications of this type of S. cerevisiae-based cellular high throughput assays will be discussed. PMID- 10838419 TI - What is the Future of High Throughput Screening? PMID- 10838418 TI - What is the Future of High Throughput Screening? PMID- 10838420 TI - Digital Imaging as a Detection Method for a Fluorescent Protease Assay in 96-Well and Miniaturized Assay Plate Formats. AB - The demand to increase throughput in HTS programs, without a concomitant addition to costs, has grown significantly during the past few years. One approach to handle this demand is assay miniaturization, which can provide greater throughput, as well as significant cost savings through reduced reagent costs. Currently, one of the major challenges facing assay miniaturization is the ability to detect the assay signal accurately and rapidly in miniaturized formats. Digital imaging is a detection method that can measure fluorescent or luminescent signals in these miniaturized formats. In this study, an imaging system capable of detecting the signal from a fluorescent protease assay in multiple plate formats was used to evaluate this detection method in an HTS environment. A direct comparison was made between the results obtained from the imaging system and a fluorescent plate reader by screening 8,800 compounds in a 96-well plate format. The imaging system generated similar changes in relative signal for each well in the screen, identified the same active compounds, and yielded similar IC(50) values as compared to the plate reader. When a standard protease inhibitor was evaluated in 96-, 384-, 864-, and 1536-well plates using imaging detection, similar IC(50) values were obtained. Furthermore, similar dose response curves were generated for the compound in 96- and 384-well assay plates read in a plate reader. These results provide support for digital imaging as an accurate and rapid detection method for high-density microtiter plates. PMID- 10838421 TI - Development of a p38 Kinase Binding Assay for High Throughput Screening. AB - p38 is a member of the mitogen-activated protein kinase (MAPK) family of serine/threonine kinases, which is activated by cellular stressors and has been shown to be a critical enzyme in the synthesis and action of proinflammatory cytokines, tumor necrosis factor-a (TNF-alpha) and interleukin-1beta (IL-1beta). A group of pyridinyl imidazole compounds such as SB202190 have been identified as selective inhibitors of p38 that bind directly to the ATP pocket of the enzyme. These compounds inhibit the p38 kinase activity, block TNF-alpha and IL-1beta secretion both in vivo and in vitro and are found to be effective in animal models of arthritis, bone resorption, and endotoxin shock. We postulated that other classes of compounds capable of competing the binding of pyridinyl imidazole with p38 enzyme could have efficacy in the treatment of inflammatory diseases. Therefore, a simple and robust assay was developed to measure the ability of small molecules to inhibit the binding of tritium-labeled pyridinyl imidazole, SB202190, to recombinant p38 kinase. For assay development, the human p38 gene was cloned in baculovirus and then expressed in insect cells. Tritiated SB202190 was synthesized and used as the p38 ligand for a competitive filter binding assay. This assay has been used successfully to screen both synthetic and combinatorial chemical libraries for other classes of p38 kinase inhibitors. PMID- 10838422 TI - Miniaturization of a Mammalian Cell-Based Assay: Luciferase Reporter Gene Readout in a 3 Microliter 1536-Well Plate. AB - The combined efforts of the fields of combinatorial chemistry and genomics have significantly increased the number of compounds and therapeutic targets available for screening. The number of compounds will reach into the million range in the near future and provide vast chemical diversity for drug discovery. However, this reservoir of chemical diversity creates downstream hurdles for any screening effort. Properly examining this number of compounds increases investments dramatically, both in the number of dollars spent and amount of limited reagents depleted. Traditional HTS techniques, such as the use of 96-well microtiter plates, have paved the way for faster processing speeds, but are being rapidly overwhelmed by screening demands. Miniaturization of such assays will allow for greater throughput, while concurrently reducing cost. To date, miniaturization efforts have been most successfully applied to bacterial and soluble protein based assays. Questions about the ability to deliver microquantities of mammalian cells without disruption of the cell membrane and/or activation of stress responses have been raised. An assay has been developed in which a human T-cell screen has been adapted to a 1536-well plate format. Through the use of a luciferase reporter gene system, it is shown that a mammalian cell-based assay may be successfully performed in 3 ul and potent inhibitors of the target of interest identified. PMID- 10838423 TI - The High Throughput Screening Infrastructure: The Right Tools for the Task. AB - HTS is a key component of pharmaceutical lead identification process. Over recent years, the pharmaceutical industry has experienced significant increases in the throughput capabilities of its HTS functions. In those companies where HTS has been effectively deployed, it is now possible to screen the entire corporate compound collection against a pharmacological target within a timescale of several weeks to a few months. This capability has been realized, not as a result of the purchase of any one particular piece of hardware, but rather through the development of a truly effective HTS infrastructure that matches the needs of the parent organization. Central to this is the need to understand how to effectively combine the use of the different types of hardware available to the HTS specialist. The use of both modular workstations and single-arm robotic systems have underpinned most HTS groups operations. Recent advances in the field of multiple-arm robotic systems and dedicated automation systems offer even further potential for increasing productivity. This article describes our experience with the use of a dedicated automation system for HTS applications. PMID- 10838424 TI - Monitoring Inositol-Specific Phospholipase C Activity Using a Phospholipid FlashPlate(R). AB - Inositol-specific phospholipase Cs(PLCs) are a group of enzymes involved in the signal transduction pathway of many plasma membrane receptor mediated events. We developed a modified solid surface to capture [(3)H] PIP(2) onto the Basic FlashPlate(R) in order to monitor PLC activity. Our results clearly demonstrate the utility of [(3)H] PIP(2)-Coated Phospholipid FlashPlate(R) microtiter plates for assessing PLC activity for HTS of receptor-coupled functional assays. The results show that PLC activity can be measured easily from a variety of sources including purified recombinant enzyme preparations, crude HL60 cell lysates and permeabilized A431 human carcinoma cells. Moreover, this format provides a surface comparable to that used for classical solution based radiolabeled mixed phospholipid micelle studies and illustrates the feasibility of this assay for measuring PLC activation in a variety of different drug screening assays. PMID- 10838426 TI - LANCEtrade mark: Homogeneous Assay Platform for HTS. PMID- 10838425 TI - Introduction LANCEtrade mark vs. HTRF(R) Technologies (or Vice Versa). PMID- 10838427 TI - HTRF(R) Technology. PMID- 10838428 TI - Rapid Solubility Determination Using Vapor-Phase Osmometry. AB - Because of the need for resource-sparing assays of the solubility of new drug candidates, we sought to develop and validate a rapid method for determining the solubility of nonvolatile pharmaceutical solids in water. Vapor-phase osmometry was used to determine the concentration of drugs in saturated solutions prepared by a rapid ultrasound-mediated dissolution protocol. The osmolality of saturated solutions as measured by the vapor-phase osmometer is an excellent predictor of the solubility of pharmaceutical solids in water. Each osmolality measurement requires less than 10 ul of saturated solution and takes less than 2 min to complete. For small-molecule drugs with solubilities greater than 10 g/kg, osmometry may prove to be a rapid and accurate method for determining the water solubilities of drugs. PMID- 10838429 TI - A Screening Strategy Based on Differential Binding of Ligand to Receptor and to Binding Proteins: Screening for Compounds Interacting with Corticotrophin Releasing Factor-Binding Protein. AB - The ligand-receptor interaction has been commonly used in development of high throughput screening assays for new drugs. In some cases, an endogenous ligand interacts not only with membrane receptors but also with soluble binding proteins. Corticotrophin-releasing factor (CRF) is an important stress neurotransmitter/hormone involved in both the central and peripheral nervous systems. CRF exerts its function by interacting with CRFR1 and CRFR2 receptors. In addition, CRF-binding protein (CRF-BP) binds CRF with high affinity. Accordingly, CRF-BP has been suggested to play an important role in modulating CRF function. Based on the potential involvement of CRF-BP in many neurological disorders, it is desirable to develop a screening assay to look for drugs that either mimic or interfere with CRF binding to CRF-BP. An assay was developed to monitor the interactions of radiolabeled CRF with human/rat CRF-BP and the mouse CRFR1 (mCRFR1) receptor. By carefully examining the binding characteristics of radiolabeled CRF to mCRFR1, the assay was able to identify compounds that bind to CRF-BP with high affinity and have little or no affinity for mCRFR1 receptors. Based on a mathematical model, we have verified the screening system with several well-characterized CRF ligands that all have different affinities for CRF receptors and CRF-BP. PMID- 10838430 TI - 3,6-Fluorescein Diphosphate: A Sensitive Fluorogenic and Chromogenic Substrate for Protein Tyrosine Phosphatases* AB - A highly sensitive and continuous protein tyrosine phosphatase (PTPase) assay using 3,6-fluorescein diphosphate (FDP) is described. Leukocyte phosphatase CD45 (leukocyte common antigen), protein tyrosine phosphatase-1B, and leukocyte common antigen-related protein LAR preferentially hydrolyze FDP to fluorescein monophosphate (FMP) with V(max) and K(m) values comparable with those of phosphotyrosine peptide substrates. Further hydrolysis of FMP to fluorescein was less efficient because of increased K(m) values compared with those of FDP. FMP absorbs strongly at 445 nm and fluoresces intensely near 515 nm, both of which are insensitive to pH perturbations above pH 6. Its high catalytic efficiency, coupled with the highly sensitive dual detection in the visible wavelength region and wider pH operating range, make FDP the substrate of choice for PTPase inhibitor screening in HTS format and assay miniaturization. PMID- 10838431 TI - Single Molecule Detection Technologies in Miniaturized High Throughput Screening: Fluorescence Correlation Spectroscopy. AB - Fluorescence assay technologies used for miniaturized high throughput screening are broadly divided into two classes. Macroscopic fluorescence techniques (encompassing conventional fluorescence intensity, anisotropy [also often referred to as fluorescence polarization] and energy transfer) monitor the assay volume- and time-averaged fluorescence output from the ensemble of emitting fluorophores. In contrast, single-molecule detection (SMD) techniques and related approaches, such as fluorescence correlation spectroscopy (FCS), stochastically sample the fluorescence properties of individual constituent molecules and only then average many such detection events to define the properties of the assay system as a whole. Analysis of single molecular events is accomplished using confocal optics with an illumination/detection volume of approximately 1 fl (10( 15) L) such that the signal is insensitive to miniaturization of HTS assays to 1 ul or below. In this report we demonstrate the general applicability of one SMD technique (FCS) to assay configuration for target classes typically encountered in HTS and confirm the equivalence of the rate/equilibrium constants determined by FCS and by macroscopic techniques. Advantages and limitations of the current FCS technology, as applied here, and potential solutions, particularly involving alternative SMD detection techniques, are also discussed. PMID- 10838432 TI - High Throughput Screening with Multiphoton Excitation. AB - Fluorescence detection is extensively used in high throughput screening. In HTS there is a continuous migration toward higher density plates and smaller sample volumes. In the present report we describe the advantages of two-photon or multiphoton excitation for HTS. Multiphoton excitation (MPE) is the simultaneous absorption of two long-wavelength photons to excite the lowest singlet state of the fluorophore. MPE is typically accomplished with short but high-intensity laser pulses, which allows simultaneous absorption of two or more photons. The intensity of the multiphoton-induced fluorescence is proportional to the square, cube, or higher power of the instantneous photon flux. Consequently, two-photon or multiphoton excitation only occurs at the focal point of the incident beam. This property of two-photon excitation allows the excited volume to be very small and to be localized in the center of each well in the HTS plate. We show that two photon-induced fluorescence of fluorescein can be reliably measured in microwell plates. We also show the use of 6-carboxy fluorescein as a pH probe with two photon excitation, and measure 4'-6-diamidino-2-phenylindole (DAPI) binding and two-photon-induced fluorescence. In further studies we measure the time-dependent intensity decays of DAPI bound to DNA and of calcium-dependent fluorophores. Finally, we demonstrate the possibility of three-photon excitation of several fluorophores, including indole, in the HTS plate. These results suggest that MPE can be used in high-density multiwell plates. PMID- 10838433 TI - Development of an In Vitro Screening Assay for Compounds that Increase Bone Formation. AB - Although parathyroid hormone (PTH) has the ability to stimulate bone growth in both rats and humans, its mechanism of action is not fully understood at the molecular level. An in vitro marker that reflects the in vivo anabolic actions of PTH would facilitate the discovery of small-molecule compounds that stimulate bone growth. We therefore compared the patterns of gene expression in three cell lines treated with PTH. The levels of c-fos, collagenase, interleukin-6 (IL-6), and collagen mRNA were determined by reverse transcription-polymerase chain reaction (RT-PCR) in three osteoblast-like cell lines. The most responsive marker was c-fos, which was induced 5-10-fold after 1 h of PTH treatment in the UMR106 01 cell line. Because it is a critical early response gene in bone growth, we investigated the possibility of using c-fos stimulation as a method to screen for compounds that can stimulate bone formation. A highly sensitive, medium throughput RT-PCR assay for c-fos mRNA expression was established using the Taqmantrade mark Detection System (Perkin Elmer, Mississauga, Ontario). Cells were treated with a series of compounds to determine the specificity of c-fos stimulation. Of the compounds tested, only PTH, prostaglandin E(2), 8-bromo-cAMP, and forskolin induced c-fos mRNA levels, indicating that this assay was specific for compounds that are known to induce cAMP and stimulate bone growth. These results indicate that a simple in vitro assay for c-fos may be a reliable method for the screening of compounds that stimulate bone growth in vivo. PMID- 10838434 TI - Stacker Modules Used in a High-Capacity Robotics System for High Throughput Screening Compound Replication. AB - High throughput screening is now established as a key component of the pharmaceutical lead identification process in many pharmaceutical companies. Over recent years, thanks to advances in assay technology, process automation, and logistics control, the throughput capacity of HTS groups has increased significantly. It is now entirely possible to screen corporate compound collections against an individual pharmacological target within a timescale of several weeks. Despite these improvements, many HTS groups find that their capacity is limited by the rate at which they can provide test compounds in a "screen-ready" format. This limitation is usually imposed by the capacity and productivity of the single-armed robotic systems utilized. We have recently constructed a robotic system aimed at overcoming this particular problem. This system uses purpose-built microplate stacker units that provide high-capacity microplate storage and, importantly, provide an easy and fast interface between the robotic system and the human operators. This paper describes this automation project and the benefits that have resulted from its deployment. PMID- 10838435 TI - What is the Future of High Throughput Screening? PMID- 10838436 TI - High Throughput Screening: Myths and Future Realities. PMID- 10838437 TI - High Throughput Screening for Drug Discovery: Continually Transitioning into New Technology. AB - Those working in HTS laboratories, pressured to find increasing numbers of drug leads while containing costs, are seeking larger compound sets, more automated systems to screen them faster, and an integrated set of equipment and consumables. Enabling technologies are continually being developed and suppliers are teaming up to supply integrated equipment and consumable sets. Miniaturization, microfluidic chips, subnanoliter dispensing, fluorescence, homogeneous assays for HTS, and virtual screening are just some of the evolving tools that HTS experts are continually evaluating and incorporating into drug discovery operations. PMID- 10838438 TI - Mechanism-Based Inhibitors: Development of a High Throughput Coupled Enzyme Assay to Screen for Novel Antimalarials. AB - Identifying potent enzyme inhibitors through a robust HTS assay is currently thought to be the most efficient way of searching for lead molecules. We have developed a HTS assay that mimics a crucial step in an essential metabolic pathway, the purine salvage pathway of the malarial parasite Plasmodium falciparum. In this assay we have used purified recombinant enzymes: hypoxanthine guanine phosphoribosyl transferase (HGPRT) and inosine monophosphate dehydrogenase (IMPDH) from the malarial parasite and the human host, respectively. These two enzymes, which work in tandem, are used to set up a coupled assay that is robust enough to meet the stringent criteria of an HTS assay. In the first phase of our screen we seem to have identified novel inhibitors that kill the parasite by inhibiting the salvage pathway of the parasite. PMID- 10838439 TI - Homogeneous Cell- and Bead-Based Assays for High Throughput Screening Using Fluorometric Microvolume Assay Technology. AB - High throughput drug screening has become a critical component of the drug discovery process. The screening of libraries containing hundreds of thousands of compounds has resulted in a requirement for assays and instrumentation that are amenable to nonradioactive formats and that can be miniaturized. Homogeneous assays that minimize upstream automation of the individual assays are also preferable. Fluorometric microvolume assay technology (FMAT) is a fluorescence based platform for the development of nonradioactive cell- and bead-based assays for HTS. This technology is plate format-independent, and while it was designed specifically for homogeneous ligand binding and immunological assays, it is amenable to any assay utilizing a fluorescent cell or bead. The instrument fits on a standard laboratory bench and consists of a laser scanner that generates a 1 mm(2) digitized image of a 100-umm deep section of the bottom of a microwell plate. The instrument is directly compatible with a Zymark Twistertrade mark (Zymark Corp., Hopkinton, MA) for robotic loading of the scanner and unattended operation in HTS mode. Fluorescent cells or beads at the bottom of the well are detected as localized areas of concentrated fluorescence using data processing. Unbound flurophore comprising the background signal is ignored, allowing for the development of a wide variety of homogeneous assays. The use of FMAT for peptide ligand binding assays, immunofluorescence, apoptosis and cytotoxicity, and bead based immunocapture assays is described here, along with a general overview of the instrument and software. PMID- 10838441 TI - HTS: Productivity or Oblivion? PMID- 10838440 TI - A Homogenous 384-Well High Throughput Screen for Novel Tumor Necrosis Factor Receptor: Ligand Interactions Using Time Resolved Energy Transfer. AB - The herpes virus entry mediator (HVEM) receptor and its ligand, HVEM-L, are involved in both herpes simplex virus type-1 (HSV-1) herpes simplex virus type-2 (HSV-2) infection, and in T-cell activation such that antagonists of this interaction are expected to have utility in viral and inflammatory diseases. In this report we describe the configuration of a homogeneous 384-well assay based on time-resolved energy transfer from a europium chelate on the HVEM receptor to an allophycocyanin (APC) acceptor on the ligand. Specific time resolved emission from the acceptor is observed on receptor:ligand complex formation. The results of various direct and indirect labeling strategies are described. Several assay optimization experiments were necessary to obtain an assay that was robust to automation and file compound interference while sensitive to the effect of potential inhibitors. The signal was stable for more than 24 h at room temperature using the Eu(3+) chelates, suggesting no dissociation of the lanthanide ion. The 384-well assay was readily automated and was able to identify more than 99.5% of known positive controls in the validation studies successfully. Screening identified both a series of known potent inhibitors and several structural classes of hits that readily deconvoluted to yield single compound inhibitors with the desired functional activity in secondary biological assays. The equivalence of the data in 384- and 1536-well formats indicates that routine implementation of 1536-well chelate-based energy transfer screening appears to be primarily limited by liquid handling rather than detection issues. PMID- 10838442 TI - HTS 2010: A Retrospective Look at Screening in the First Decade of the New Millennium. PMID- 10838443 TI - Faculty or Factory? Why Industrializing Drug Discovery is Inevitable. PMID- 10838444 TI - Time-Resolved Fluorescence Energy Transfer DNA Helicase Assays for High Throughput Screening. AB - DNA helicases are responsible for the unwinding of double-stranded DNA, facilitated by the binding and hydrolysis of 5'-nucleoside triphosphates. These enzymes represent an important class of targets for the development of novel anti infective agents particularly because opportunity exists for synergy with existing therapies targeted at other enzymes involved in DNA replication. Unwinding reactions are conventionally monitored by low throughput, gel-based radiochemical assays; to overcome the limitations of low throughput to achieve comprehensive characterization of adenosine triphosphate (ATP)-dependent unwinding by viral and bacterial helicases and the screening for unwinding inhibitors, we have developed and validated homogeneous time-resolved fluorescence energy transfer (TRET) assays. Rapid characterization and screening of DNA helicase has been performed in 96- and 384-well plate densities, and the ability to assay in 1536-well format also demonstrated. We have successfully validated and are running full high throughput runs using 384-well TRET helicase assays, culminating in the identification of a range of chemically diverse inhibitors of viral and bacterial helicases. For screening in mixtures, we used a combination of quench correction routines and confirmatory scintillation proximity (SP) assays to eliminate false-positives due to the relatively high levels of compound quenching (unlike other Ln(3+)-based assays). This strategy was successful yet emphasised the need for further improvements in helicase assays. PMID- 10838445 TI - Analysis of Apparent Noncompetitive Responses to Competitive H(1)-Histamine Receptor Antagonists in Fluorescent Imaging Plate Reader-Based Calcium Assays. AB - We have examined the utility of high throughput fluorescent imaging plate reader (FLIPR)-based calcium assays for pharmacological characterization of G-protein coupled receptors (GPCRs) using recombinant and native human H1-histamine receptors (H(1)-HR), expressed in HEK293 and HeLa S3 cells, respectively, as model systems. For stably transfected HEK293 cell lines, the potency of histamine for elevating intracellular calcium increased (pD(2), 7.13 and 7.86) with increased H(1)-HR density (about 0.8 and 14 pmol/mg protein, respectively), though histamine binding affinities were similar. The classic H(1)-HR competitive antagonists diphenhydramine and chlorpheniramine appeared noncompetitive by causing depressions of the maximal histamine responses along with rightward shifts of histamine concentration-response curves, thus precluding Schild analysis. Applying the generalized Cheng-Prusoff equation to antagonist concentration-response curves for inhibition of fixed histamine concentrations yielded apparent pK(b) values that were consistent among recombinant and native receptors at different expression levels. These pK(b) values for diphenhydramine and chlorpheniramine (e.g., 7.83 and 8.77, respectively) were in good agreement with binding pK(i) values (e.g., 7.98 and 8.52, respectively). Apparent antagonist affinities determined from FLIPR calcium and competition binding assays were also consistent for the competitive antagonists mepyramine, tripelennamine, and promethazine. In phosphoinositide hydrolysis assays, chlorpheniramine exhibited insurmountable inhibition of histamine calcium responses, although to a lesser extent than that observed in calcium assays; pK(b) values were similar. These results demonstrate that competitive antagonist potencies can be attained from FLIPR-derived data by application of the generalized Cheng-Prusoff equation, despite apparent noncompetitive antagonism under these assay conditions. Apparent noncompetitive antagonist effects may in part be attributable to a lack of equilibrium of histamine and antagonists with H(1)-HR within the short duration of rapid transient effects of histamine on intracellular calcium. PMID- 10838446 TI - Toxicity Screening of a Combinatorial Library: Correlation of Cytotoxicity and Gene Induction to Compound Structure. AB - Combinatorial chemistry has increased the number of compounds available for efficacy and safety assessment by several orders of magnitude and has made high throughput assays essential. To test whether higher throughput toxicity assays could be of utility in screening compounds in early development, a selected set of combinatorial chemistry compounds was screened for induction of 70-Kd heat shock protein (HSP70) and 45-Kd growth arrest and DNA damage protein (GADD45) mRNA levels as well as cytotoxicity, in HepG2 cells, using a 96-well microtiter plate format. Both assays, the branched DNA (Quantigene) assay for mRNA levels and MTT for cytotoxicity, were robust enough to be incorporated into a screening format using a single replicate and a single concentration of compound. Significantly, a structure/toxicity correlation was established with this set of compounds with cytotoxicity and gene induction patterns linked to compound structure. Therefore, this type of early screening may be useful in identifying toxic substituents, enabling the design of libraries with less potential for toxicity. While structure/toxicity correlations were observed, no relationship was observed between GADD45 gene induction and mutagenesis as measured by the Ames bacterial reverse mutation assay. PMID- 10838447 TI - Amphibian Melanophore Technology as a Functional Screen for Antagonists of G Protein Coupled 7-Transmembrane Receptors. AB - Xenopus laevis melanophores stably expressing 7-transmembrane G-protein-coupled receptors were established and evaluated, either as a primary screening utility for antagonists of the human calcium receptor, or as a screen to assign function to binding inhibitors of human cannabinoid receptors. Stably or transiently expressing melanophores responded selectively to respective effectors of the human calcium, cannabinoid, and neurokinin-1 receptors. Several selective cannabinoid receptor-binding inhibitors of known potency were characterized as agonists or antagonists of the human peripheral cannabinoid (CB(2)) receptor. The results were consistent with changes in cAMP content of hCB(2)-transfected human embryonic kidney (HEK) cells challenged with the same CB(2)-binding antagonists. A stable melanophore cell line expressing the human calcium receptor was used to screen a compound collection directly for functional antagonists, several of which were confirmed as antagonists in secondary screens by stimulating parathyroid hormone (PTH) secretion from bovine parathyroid cells. The percentage of hits in this cell-based screen was reasonably low (1.2%), indicating minimal interference due to toxic effects and validating melanophores as a primary screening modality. Also described is the development of a novel procedure for cryopreservation and reconstitution of cells retaining functional human receptors. () PMID- 10838448 TI - Epsilon-aminocaproic acid inhibits the activity of factor VIII inhibitors in patients with severe haemophilia A in vivo and in vitro. AB - Haemophilia patients with inhibitors pose a formidable challenge for patient management. This is particularly problematic in developing countries, where porcine factor VIII, FEIBA, factor VIIa or immunoadsorption column are generally unavailable or unaffordable. Under these circumstances, any effective modality of affordable treatment is welcome. We investigated, both in vivo and in vitro, the effect of epsilon-aminocaproic acid (EACA) on the inhibitory activity of factor VIII inhibitor. It was found that in vitro EACA (final concentration 1.25-5 mg/ml) substantially inhibited the activity of the inhibitors, while the same concentration of EACA had no effect on other immunological reactions like red cell agglutination and immunofluorescence. The inhibitory action of EACA on factor VIII inhibitor was also confirmed in an improvised antigen-binding ELISA system. Further, the inhibitory activity of EACA was confirmed in 2 patients, in whom the inhibitory activity persisted for 15 min following infusion of EACA (100 mg/kg over 10 min). EACA was found to be even more effective in local wound application in patients of haemophilia A with inhibitors. EACA at the concentration cited did not act as an inhibitor of factor VIII inhibitor through occupancy of lysine binding sites. The inhibitory activity of EACA on factor VIII inhibitor was equally seen with recombinant factor VIII also; hence this action cannot be explained by its antifibrinolytic activity. PMID- 10838449 TI - Possible association between cytomegalovirus infection and gastrointestinal bleeding in hemophiliac patients. AB - Cytomegalovirus (CMV) infection is of major concern in immunocompromised and immunosuppressed patients. Prior to the introduction of HIV-1 antibody screening and efficient virucidal processes to inactivate viruses, individuals with a factor VIII or factor IX deficiency had a high risk of contracting HIV-1 infection through the infusion of contaminated blood products. In addition, blood products were also frequently associated with alterations in immune function. This study investigated the frequency of active CMV infection and its clinical relevance in Brazilian hemophiliacs. One hundred hemophiliacs were screened for the presence of CMV-DNA in their blood using nested PCR. Twenty-five out of 100 patients (25%) were positive for CMV-DNA and 24 of these 100 patients (24%) were HIV-1 positive; 6 of these 24 (25%) were positive for CMV-DNA. A similar frequency was observed among HIV-1-negative patients. In 60 hemophiliacs, the clinical relevance of the CMV infection was assessed. Twenty-one patients were positive for CMV-DNA. Of these, 10 had gastrointestinal bleeding compared to only 9 of 39 patients who were CMV-DNA negative (p = 0.05; chi(2) test). These data indicate a high prevalence of active CMV infection in Brazilian hemophiliac patients, irrespective of whether the patients were or were not infected by HIV 1. There was a possible association between the presence of CMV and the occurrence of gastrointestinal bleeding. PMID- 10838450 TI - Basic fibroblast growth factor modulates the surface expression of effector cell molecules and primes respiratory burst activity in human neutrophils. AB - Basic fibroblast growth factor (b-FGF) mediates a variety of biological responses such as angiogenesis and hematopoiesis. We examined the effect of b-FGF on human neutrophil functions in vitro. The surface expression of effector cell molecules on neutrophils was determined by flow cytometry and monoclonal antibodies. b-FGF increased the expression of CD11b leukocyte integrin and complement receptor type 1 on neutrophils and decreased the expression of L-selectin on neutrophils in a dose- and time-dependent manner. We also examined the effect of b-FGF on the respiratory burst activity in neutrophils. Although b-FGF alone did not induce intracellular oxidative product formation by neutrophils, it enhanced H(2)O(2) production in neutrophils stimulated by N-formyl-methionyl-leucyl-phenylalanine or phorbol myristate acetate. These findings suggest that b-FGF may participate in the inflammatory process via modulating the surface expression of effector cell molecules and enhancing respiratory burst activity in neutrophils. PMID- 10838451 TI - Differential diagnosis of Hb EE and Hb E-beta(0)-thalassemia by protein and DNA analyses. AB - DNA analysis was used to confirm the Hb EE genotype and to differentiate from the possible genotype of Hb E-beta(0)-thalassemia in two Malay patients. The first patient was a 13-year-old Malay girl, whose parents were available for family studies. The second patient was a 69-year-old Malay woman with no living family members. The presence of Hb E in both propositi was confirmed by: (1) its characteristic electrophoretic mobilities on alkaline/acid gels; (2) its chromatographic properties on anion/cation exchangers, and (3) its mildly insoluble properties. However, differential diagnosis of Hb EE and Hb E-beta(0) thalassemia was challenging in these two cases. In the former, this was because of the possible interactions of the parents' phenotypes; i.e., the mother had a similar phenotype. In the latter, it was due to the lack of any living family members for family studies. In this communication, we present the protein and DNA analyses, including data on the use of the restriction enzyme Mnl I, for the definitive diagnosis of the Hb EE genotype in the propositi of these two Malay families. PMID- 10838452 TI - Evidence of lymphocyte alveolitis by bronchoalveolar lavage in thalassemic patients with pulmonary dysfunction. AB - Pulmonary dysfunction represents one of the least studied complications in thalassemic patients. Probably, it is due to the absence of pulmonary symptoms. There are few works in the literature, and contradictory results have been published. The aim of this study was to define the spirometric pattern and the possible causes of lung impairment by testing bronchoalveolar lavage (BAL) with pathological pulmonary function tests (PFTs). Furthermore, diffusion capacity tests for carbon monoxide corrected for hemoglobin value (Dco*) were performed. We studied 48 thalassemic patients (27 F and 21 M), with an age range from 8 to 23 years, divided into two groups on the basis of PFTs results. Thus, group A was formed by 16 patients with restrictive spirometric patterns of whom 14 had also reduced Dco* values and group B consisted of 32 patients with normal PFTs and Dco* values. Patients of group A underwent chest high-resolution computing tomography (CHRCT) and BAL whose fluid was analyzed by microbiologic and cytological assays. A pathological CHRCT picture was present in 8 patients. Nine out of 16 patients who accepted to undergo BAL had a chronological age greater than 17 years with a mean bone age of 13.9 years. BAL results showed lymphocyte alveolitis in 6 patients and a normal cytogram in 3, while alveolar iron-laden macrophages were present in 4 out of 6 patients with alveolitis and 2 out of 3 patients with normal cytogram. Moreover, all examined BAL fluids showed a normal CD4/CD8 ratio, while only 2 patients showed an altered serum CD4/CD8 ratio. We demonstrated the presence of (1) lung-restrictive syndrome in 16 of the oldest thalassemic patients; (2) lymphocyte alveolitis in 6 patients, and (3) a picture of interstitial fibrosis by CHRCT in 8 of them. All these data are suggestive of a diagnosis of interstitial lung disease secondary to thalassemia. BAL helped to identify the presence of alveolar iron-laden macrophages that represented a local defense mechanism against free iron. This latter finding therefore might be the primary cause of the lung impairment promoting an oxidative damage. Further studies are needed to investigate this hypothesis and therapeutical potentials. PMID- 10838453 TI - Type I and type II T cell profiles in chronic myelogenous leukemia. AB - T cell immunity is considered to play an important role in the control of cell growth in chronic myelogenous leukemia (CML) although information regarding the characteristics of T lymphocytes in CML patients is limited. Using flow cytometric analysis of intracellular cytokine expression at the single-cell level, we analyzed helper T and cytotoxic T subsets in 8 CML patients. The percentage of interferon-gamma (IFN-gamma) single-positive CD4 cells (Th1) and that of interleukin-4 (IL-4) single-positive CD4 cells (Th2) was markedly decreased in pretreated CML patients compared to normal controls. In addition, the percentage of IFN-gamma/IL-4 double-positive CD4 cells (Th0) was also reduced. Consequently, the percentage of IFN-gamma/IL-4 double-negative CD4 cells was markedly increased. Similarly, a reduction in IFN-gamma single-positive CD8 cells (Tc1) and IFN-gamma/IL-4 double-positive CD8 cells (Tc0) and an increase in IFN-gamma/IL-4 double-negative CD8 cells were observed in pretreated CML patients. Imbalance of these parameters was markedly improved following cytoreduction therapy. Our findings directly indicate anergic states in CML patients. Determination of the factors that affect Th and Tc profiles may lead to further understanding of immunological states and the development of effective immunotherapy. PMID- 10838454 TI - Diagnosis of granulocytic sarcoma by fine-needle aspiration cytology. AB - Granulocytic sarcoma (GS) occurs in patients with chronic myeloproliferative disorders as well as in patients with acute myeloid leukemia (AML). These tumorous masses can occur anywhere in the body and have to be differentiated from lymphoma, carcinoma or infectious processes. We report the results of fine-needle aspiration cytology (FNAC) in 26 patients with GS. Seventeen patients suffered from AML and 9 from chronic myeloid leukemia (CML) blast crisis. In 5 patients with AML, GS was the initial presentation of hematological malignancy, in the remaining 21 patients, FNAC confirmed relapse of AML or extramedullary blast crisis of CML. In 8 patients, GS was located in the skin, in 17 in the lymph node and in another patient in the spinal canal. This study demonstrates the clinical utility and diagnostic accuracy of FNAC in the evaluation of GS from multiple sites. PMID- 10838455 TI - New rearrangement pattern after treatment of hairy-cell leukemia with 2 chlorodeoxyadenosine. AB - Leukemic hairy cells are clonally proliferating B-lymphoid cells with clonal rearrangements of genes for immunoglobulin chains. We describe a patient with a new hairy-cell clone after treatment with 2-chlorodeoxyadenosine (2-CdA). In this patient, a single course of 2-CdA resulted in good partial remission of hairy cell leukemia, but Southern blot analysis of bone marrow biopsies and polymerase chain reaction using seminested amplifications with consensus primers revealed a new rearranged band 4 months after therapy with 2-CdA. Four years after therapy, the patient is in complete clinical remission and both bands disappeared during follow-up. The new rearranged band might have been related to prior treatment of hairy-cell leukemia with 2-CdA. PMID- 10838456 TI - Mycoplasma-pneumoniae-induced thrombotic thrombocytopenic purpura. AB - Thrombotic thrombocytopenic purpura (TTP) is a fatal disease characterized by widespread platelet aggregation, hemolytic anemia and fever with renal and neurological involvement. Different factors have been associated with the development of TTP, e.g. infections, pregnancy, chemotherapy, drug therapy and bone marrow transplantation. Recent data imply that all these different causes may induce the disease by decreasing the activity of the plasma von Willebrand factor-cleaving protease resulting in unusually large von Willebrand factor multimers that later on initiate the cascade of TTP. In this communication, we present a unique association between infection with Mycoplasma pneumoniae and TTP. We believe that the emergence of antibodies that cross-react with Mycoplasma and the protease might elucidate in this case the pathogenesis of TTP. PMID- 10838457 TI - Secondary cutaneous infiltration in B cell chronic lymphocytic leukemia. AB - We describe a patient presenting with B cell chronic lymphocytic leukemia (B-CLL) who subsequently developed cutaneous infiltrates. Specimens of the blood, bone marrow and cutaneous infiltrations all showed the same heavy-chain gene rearrangement. Following failure of conventional chemotherapy, and in view of the similarity of the disease to cutaneous T cell lymphoma, interferon-alpha therapy was employed with satisfactory results. Introduction of this cytokine to the therapeutic modalities for secondary cutaneous B-CLL would hopefully change the poor outcome of this entity, or at least could produce a better quality of life. Loss of histidine decarboxylase activity in the infiltrating cells - in contrast to circulating lymphocytes - may be associated with the transformation of B-CLL to a more aggressive infiltrative form, offering a possible explanation for tissue invasiveness. The changing character of the disease raises the possibility of a second mutational event in the course of B-CLL. PMID- 10838459 TI - Elevation of factor VIII during decreased oxygen pressure. PMID- 10838458 TI - Clinical diversity of sickle cell disease in western india - influence of genetic factors. PMID- 10838460 TI - Growth in human milk-Fed very low birth weight infants receiving a new human milk fortifier. AB - BACKGROUND/AIMS: Human milk fortification has been advocated to enhance premature infants' growth. We, therefore, undertook this study of a new human milk fortifier containing more protein than a reference one. METHODS: Open, randomized, controlled, multiclinic trial, with weekly growth parameters and safety evaluations in premature infants <1,500 g. RESULTS: The 2 groups did not differ in demographic and baseline characteristics. The adjusted daily milk intake was significantly higher in the infants fed reference human milk fortifier (n = 29; 154.2 +/- 2.1 vs. 144.4 +/- 2.5 ml/kg/day, mean +/- SE; p < 0.05). Both human milk fortifiers produced increases over baseline in weight, length, and head circumference, with greater gains observed in the new human milk fortifier fed infants for the former two parameters (weight gain 26.8 +/- 1.3 and 20.4 +/- 1.2 g/day, p < 0.05; head circumference 1.0 +/- 0.1 and 0.8 +/- 0.1 cm/week; length 0.9 +/- 0.1 and 0.8 +/- 0.1 cm/week, respectively). Serum chemistries were normal and acceptable for age. Study events were typical for premature infants and similar in both groups. CONCLUSIONS: This new human milk fortifier had comparable safety to the reference human milk fortifier and promoted faster weight gain and head circumference growth. PMID- 10838461 TI - Effect of glycemic control and vitamin E supplementation on total glutathione content in non-insulin-dependent diabetes mellitus. AB - Thirty patients with non-insulin-dependent diabetes mellitus were selected for the study. 15 age-matched healthy volunteers served as controls. Serum malonaldehyde, total glutathione, and vitamin E levels were estimated before and after glycemic control and after 4 weeks of vitamin E supplementation. Both total glutathione and vitamin E levels increased after glycemic control and showed an increase after vitamin E supplementation. Malonaldehyde levels lowered after glycemic control, but remained higher than controls. Since vitamin E supplementation significantly decreased oxidative stress in the present study, it may play a role in reducing free-radical-induced oxidant injury in diabetes mellitus. PMID- 10838462 TI - Nutrient intake and growth of preschool children from different socioeconomic regions in the city of oaxaca, Mexico. AB - Dietary intake and anthropometric measurements were determined in 124 preschool children, aged 4-6 years, living in the city of Oaxaca, Mexico. The children were classified by socioeconomic status, depending upon their living conditions and family income. A composite diet representing the typical diet according to socioeconomic group was formulated and chemically evaluated. Corn tortillas and beans were the primary protein and energy sources for the children of this study. The inclusion of dairy products, animal products, and fruits and vegetables was not common. The children in the low-income group, which represents the majority of the children of this area, received only 63% of the recommended intake. Also for the low-income group, protein and iron recommendations were met but by plant based sources. Protein contributed 10% of the energy, 78% were from carbohydrates and 12% from fat. Calcium intake was low, meeting only 34% of the recommendation and zinc intake was 51%. Fiber intake was very high, coming primarily from beans and tortillas. In the low- income group, Z scores for height and age were -1.60 for boys and -1.75 for girls. Weight-for-age Z scores were -0.90 and -0.97 for boys and girls, respectively. The weight-for-height Z scores for this group were 0.33 and 0.12 for boys and girls, respectively. Although energy needs were not met, extreme stunting was not detected. PMID- 10838463 TI - Effect of a hypocaloric diet, increased protein intake and resistance training on lean mass gains and fat mass loss in overweight police officers. AB - We compare the effects of a moderate hypocaloric, high-protein diet and resistance training, using two different protein supplements, versus hypocaloric diet alone on body compositional changes in overweight police officers. A randomized, prospective 12-week study was performed comparing the changes in body composition produced by three different treatment modalities in three study groups. One group (n = 10) was placed on a nonlipogenic, hypocaloric diet alone (80% of predicted needs). A second group (n = 14) was placed on the hypocaloric diet plus resistance exercise plus a high-protein intake (1.5 g/kg/day) using a casein protein hydrolysate. In the third group (n = 14) treatment was identical to the second, except for the use of a whey protein hydrolysate. We found that weight loss was approximately 2.5 kg in all three groups. Mean percent body fat with diet alone decreased from a baseline of 27 +/- 1.8 to 25 +/- 1.3% at 12 weeks. With diet, exercise and casein the decrease was from 26 +/- 1.7 to 18 +/- 1.1% and with diet, exercise and whey protein the decrease was from 27 +/- 1.6 to 23 +/- 1.3%. The mean fat loss was 2. 5 +/- 0.6, 7.0 +/- 2.1 and 4.2 +/- 0.9 kg in the three groups, respectively. Lean mass gains in the three groups did not change for diet alone, versus gains of 4 +/- 1.4 and 2 +/- 0.7 kg in the casein and whey groups, respectively. Mean increase in strength for chest, shoulder and legs was 59 +/- 9% for casein and 29 +/- 9% for whey, a significant group difference. This significant difference in body composition and strength is likely due to improved nitrogen retention and overall anticatabolic effects caused by the peptide components of the casein hydrolysate. PMID- 10838464 TI - Long-term oral beta-carotene supplementation in patients with cystic fibrosis - effects on antioxidative status and pulmonary function. AB - To investigate the efficacy of long-term oral beta-carotene supplementation for optimizing the antioxidant status and pulmonary function in patients with cystic fibrosis (CF), 24 patients (aged 12. 8 +/- 6.3 years) were randomized to a CF supplementation or to a CF placebo group. As controls 14 healthy age-matched subjects (aged 14. 7 +/- 6.2 years) were studied. Patients of the CF supplementation group received 1 mg beta-carotene/kg body weight (BW)/day (maximally 50 mg beta-carotene/day) for the first 12 weeks; during the following 12 weeks, dosage was reduced to 10 mg beta-carotene/day. At study entry, plasma beta-carotene concentrations were significantly lower in CF patients than in controls (p < 0.001). In the CF supplementation group, plasma beta-carotene concentrations were significantly increased (baseline: 0.08 +/- 0.04 micromol/l) at the end of high-dose treatment (12th week; 0.6 +/- 0.4 micromol/l; p < 0.001), but decreased again during supplementation with 10 mg beta-carotene/day to 0.3 +/ 0.2 micromol/l at the end of the study (p < 0.001). beta-Carotene supplementation did not affect plasma concentrations of other carotenoids and retinol, but an increase in plasma alpha- and gamma-tocopherol concentrations was noticed. During high-dose treatment, a significant decrease in TBA-MDA complexes and a correction of total antioxidative capacity was observed. During the treatment, pulmonary exacerbation could be corrected significantly (p < 0.05). We conclude that CF patients can be efficiently supplemented with 1 mg beta carotene/kg BW/day (maximally 50 mg beta-carotene/day) to achieve plasma concentrations of healthy control subjects and to minimize oxidative stress, improving the quality of life of CF patients. PMID- 10838465 TI - Dietary n-3 polyunsaturated fatty acid and status of immunocompetent cells involved in innate immunity in female rats. AB - The aim of this study was to estimate the contributions of dietary n-3 polyunsaturated fatty acid (PUFA), a representative dietary immunosuppressant, to the activity of both alveolar macrophages (AM) and natural killer (NK) cells, and compare them to those of n-6 PUFA. Twelve 5-week-old female Sprague-Dawley rats were divided into two dietary groups, one fed a 10% fat diet for 9 weeks enriched with n-3 PUFA (n-3 diet) and the other an n-6 PUFA (n-6 diet). AM reduced the release of nitric oxide, monocyte chemoattractant protein 1 and tumor necrosis factor alpha in the rats fed the n-3 diet, compared with rats fed the n-6 diet. NK cell activity was reduced by consumption of the n-3 diet. This study suggests that consumption of n-3 PUFA can ameliorate pulmonary inflammatory disorders which are affected by the reduction of not only proinflammatory cytokines but also chemokine released from AM. PMID- 10838466 TI - Artificial cell biotechnology for medical applications. AB - Artificial cells are prepared in the laboratory for medical and biotechnological applications. The earliest routine clinical use of artificial cells is in the form of coated activated charcoal for hemoperfusion. Implantation of encapsulated cells are being studied for the treatment of diabetes, liver failure and the use of encapsulated genetically engineered cells for gene therapy. We recently found that daily orally administered artificial cells containing a genetically engineered microorganism can lower the elevated urea level in uremic rats to normal levels and increase the survival of the animal. Furthermore, this can remove potassium, phosphate, uric acid and other waste metabolites from uremic plasma. Blood substitutes based on modified hemoglobin are already in phase-III clinical trials in patients with as much as 20 units infused into each patient during trauma surgery. Artificial cells containing enzymes are being developed for clinical trials in hereditary enzyme deficiency diseases and other diseases. Artificial cells are also being investigated for drug delivery and other uses in biotechnology, chemical engineering and medicine. PMID- 10838467 TI - Plasma amino acid profile on nocturnal hemodialysis. AB - BACKGROUND/AIMS: The purpose of this study was to examine the effect of nocturnal hemodialysis (NHD) on serum amino acid (AA) profile. METHODS: In a cross-over trial, we studied the AA profile in 11 patients who were switched from conventional hemodialysis to NHD. Plasma levels of AA were measured at monthly intervals for a period of 1 year. RESULTS: Concentrations of AA isoleucine, threonine, phenylalanine, valine, glycine, glutamine and taurine increased on switch to NHD. Total AA (2,907.2 +/- 542.2 vs. 3,870.2 +/- 715.2 micromol/l, p = 0.03), essential AA (EAA, 842.2 +/- 259.4 vs. 1,125. 8 +/- 235.3 micromol/l, p = 0.03), nonessential AA (NEAA, 2,307.2 +/- 399.1 vs. 2,879.5 +/- 581.2 micromol/l, p = 0.03), and branched chain AA (315.5 +/- 90.8 vs. 386.0 +/- 114.3 micromol/l, p = 0.05) concentrations were significantly higher on NHD. However, abnormalities in the ratios of EAA/NEAA, valine/glycine and tyrosine/phenylalanine persisted during NHD. CONCLUSIONS: NHD alleviated some abnormalities in the aminogram of uremia, but other metabolic derangements lingered. PMID- 10838468 TI - Measurement of fluid volume shifts during hemodialysis by A-mode ultrasonography. AB - BACKGROUND: The rate of intercompartmental fluid volume changes during hemodialysis (HD) is a major determinant of dialysis-induced hypotension and lacks direct monitoring. The aim of the study was to evaluate the feasibility of tissue thickness (TT) measurement in monitoring the mobilization of interstitial fluids during HD. METHODS: We studied the intradialytic changes in forehead TT and inferior vena cava diameter (IVCD) in 20 patients. Plasma refilling was calculated from changes in hematocrit (Hct) and ultrafiltration rates. RESULTS: During ultrafiltration of 2,437 +/- 117 ml (mean +/- SEM), Hct increased significantly from 27.9 +/- 0.7 to 30.0 +/- 0.9%. IVCD decreased significantly from 9.7 +/- 0.2 to 6.1 +/- 0.4 mm/m(2). We found a simultaneously pronounced reduction in TT from 4. 46 +/- 0.12 to 3.78 +/- 0.12 mm (> or =15.3%) with a significant correlation to plasma refilling (0.613). CONCLUSION: Volume changes in the peripheral shell tissues during HD can be monitored directly and noninvasively by A-mode ultrasound. PMID- 10838469 TI - Systematic monitor disinfection is effective in limiting HCV spread in hemodialysis. AB - BACKGROUND: Patients on chronic hemodialysis are at high risk of HCV infection due to nosocomial transmission. The strict adhesion to universal precautions is the first step in prevention, but other simple tools such as systematic monitor disinfection and the use of separate machines for anti-HCV-positive patients need to be evaluated. METHODS: A 5-year prospective study was carried out in 4 dialysis centers enrolling 135 patients. General precautions were adopted, but anti-HCV-positive patients were not isolated. In period A, lasting 24 months, monitor disinfection was performed after each dialysis session with sodium hypochlorite; peracetic acid was also used 3 times a week. In period B, lasting 36 months, 3 dialysis units (77 patients) prolonged the same preventive protocol of period A, while another unit (58 patients) also adopted the use of separate machines for anti-HCV-positive subjects. A third-generation ELISA anti-HCV test was performed every 2 months throughout the study. RESULTS: Anti-HCV antibodies were initially detected in 43 patients (31.8%), prevalence rate ranging from 25 to 39.4%. One seroconversion occurred in period A, with an overall seroconversion rate of 0.54%/year. Also in period B one seroconversion occurred (unit 2), seroconversion rate of 0.36%/year. Therefore the mean seroconversion rate throughout the 5 years was 0.43%/year. CONCLUSION: Systematic monitor disinfection may be a simple and quite effective tool to avoid nosocomial transmission of HCV infection in the hemodialysis setting. In our opinion its use is mandatory. The use of separate machines for anti-HCV-positive patients seems unnecessary. PMID- 10838470 TI - Soluble adhesion molecules and cytokines in patients with myasthenia gravis treated by plasma exchange. AB - Plasma exchange (PE) is an effective therapeutic method used in patients with myasthenia gravis (MG) refractory to common therapy and/or with life-threatening respiratory complications. Except for acetylcholine receptor antibodies (AChRAbs), some other inflammatory mediators possibly activated in MG may also be removed during PE. Serum levels of soluble adhesion molecules (sICAM-1 and sVCAM 1), IL-6 and soluble receptors for IL-2 (sIL-2R), IL-6 (sIL-6R) and TNF alpha (sTNF-R II) were measured in 20 MG patients assigned to treatment with PE. On the basis of the serum levels of AChRAb the patients were subdivided into 2 groups (8 patients with low AChRAb, 12 patients with high AChRAb). Soluble adhesion molecules and cytokines were measured before the first and last PE, at the end of the first PE and in the samples of plasma filtrate obtained during the first PE. Before the first PE patients with MG had higher serum levels of sICAM-1, sVCAM-1, sIL-2R and sTNF-R II than controls. Both after the first PE and during the course of PE, a substantial decrease in serum levels of AChRAb, sICAM-1 and sVCAM-1 was recorded. However, serum levels of sIL-2R and sTNF-R II were not significantly influenced by either a single treatment or during the course of PE. There were high levels of AChRAb, soluble adhesion molecules and soluble cytokine receptors in plasma filtrates too. Patients with high circulating AChRAb had higher serum levels of sICAM-1 and sVCAM-1 than patients with low AChRAb. Increased serum levels of soluble adhesion molecules and soluble cytokine receptors in patients with MG suggest some systemic activation of the immune response which is more pronounced in patients with high circulating AChRAb. PE led to the decrease in serum AChRAb and soluble adhesion molecules due to their effective filtration but, on the other hand, serum levels of soluble cytokine receptors were not influenced by PE, in spite of their effective filtration which is probably counteracted by their increased production, possibly stimulated by the contact of the blood with the synthetic membrane. PMID- 10838471 TI - Leukocyte sequestration in isolated guinea pig lungs during extracorporeal circulation: effects on microvascular function. AB - Neutrophils accumulate in patient lungs during clinical hemodialysis and in isolated blood-perfused guinea pig lungs due to the contact between blood and extracorporeal system. However, it is unclear how these sequestered and partly activated neutrophils affect the lung microvasculature. We, therefore, studied pulmonary vascular resistance, vascular permeability, gas exchange, and oxygen free radical production in isolated guinea pig lungs during perfusion with whole blood containing partly 'activated' neutrophils in comparison with perfusions using leukopenic blood. We also connected a Cuprophan hemodialysis membrane to the whole-blood perfusion system in order to investigate whether a dialyzer, which may further activate leukocytes, affects lung microvascular permeability, vascular resistances, and reactive oxygen species production. The sequestered neutrophils did not seem to markedly affect the lung microvascular function, since neither the leukocyte-free perfusion nor the hemodialysis membrane altered any of the measured variables as compared with whole-blood perfusion in a system without a dialyzer. We conclude that neutrophils, whether activated by a perfusion system or by a dialysis membrane, can accumulate in isolated lungs without adversely affecting the microvascular function. PMID- 10838473 TI - Long-term changes of plasma trace element concentrations in chronic hemodialysis patients. AB - BACKGROUND: Hemodialysis (HD) patients are at risk of developing trace element imbalances. METHODS: The 12 trace elements Cd, Co, Cs, Cu, La, Mg, Mo, Pb, Rb, Sr, Tl and Zn were determined in the plasma (n = 52) of 6 chronic HD patients before and after HD sessions by inductively coupled plasma mass spectrometry. Plasma trace element concentrations were monitored for 6 months. Baseline data have been compared to the concentrations at the end of the observation period to identify a potential reduction or accumulation of trace elements in HD patients. RESULTS: Plasma Cd, Co and Pb levels were about 10 times higher than in healthy adults. Concentrations of Co and Pb increased during HD sessions, whereas plasma Co and Cd increased during the study period of 6 months. Plasma Cs, Mg, Mo and Rb continuously decreased in all patients. For plasma Cu and Zn, a statistically significant rise of their plasma concentrations during HD and during the period of 6 months could be established. Concentrations of La and Tl did not change distinctly. CONCLUSION: This study revealed that plasma trace element concentrations in HD patients are distinctly different compared to that of healthy adults. Elements such as Cs, Mg, Mo and Rb are reduced and Cd, Co and Pb are accumulated in HD patients. Further studies are needed to elucidate the clinical impact of these trace element imbalances. PMID- 10838472 TI - One-year immunological evaluation of chronic hemodialysis in end-stage renal disease patients. AB - BACKGROUND: Much research has been devoted to the determination of acute leukocyte activation as well as acute cytokines production during and after blood hemodialysis membrane interaction. In contrast, few studies deal with chronic immunological evaluation of T-cell activation markers in hemodialysis. METHODS: We evaluated different immune parameters using a modified cellulose low-flux hemophan vs. synthetic high-flux polyamide membrane during 1 year in 35 stable chronic hemodialysis patients. Leukocyte counts, lymphocyte subpopulations, T cell activation markers (CD69, CD25, HLA-DR, CD54, CD62L, CD45RO, CD11a, CD28), complement-activation products (C3a) and serum elastase were measured at 0, 3, 6 and 12 months in the two patient groups and compared to 13 healthy control subjects. RESULTS: Over dialysis time, all patients showed a significant level elevation of CD69/CD3 (p < 0.005) and CD25/ CD3 (p < 0.005) phenotypes. In contrast, HLA-DR and CD45RO remained unchanged suggesting a truncated pattern of activation. T lymphocyte subset analysis showed in both hemodialyzed groups a significant decrease in the expression of CD54 (ICAM-1) when compared to controls (p < 0.005). C3a and elastase measurements showed a significant upward trend with dialysis time in both hemodialyzed groups. CONCLUSION: Although the immunological changes seen in chronic hemodialyzed patients must be interpreted in conjunction with their basal uremic states and the membrane permeability properties, our study suggests that 1-year immunological evaluation of hemodialysis membranes biocompatibility is associated with changes in the pattern of chronic T-cell activation, which is in part related to the use of a particular membrane type. Moreover, some key molecules (CD54) are affected in patients with end-stage renal disease undergoing hemodialysis. PMID- 10838474 TI - Haematopoietic progenitors from umbilical cord blood. AB - BACKGROUND/AIM: The aim of this study is to improve the obstetrician-based cord blood collection system and an efficient recovery of CD34+ haematopoietic progenitor stem cells. METHODS: CD34+ cells were purified from total blood using a positive selection enrichment method, called Mini-Macs. RESULTS: The final yield of CD34+ cells we obtained was 10(4) cells/ml, with a CD34+ purity of 99%. CONCLUSION: Our results confirm that, by using this method, it is possible to get a significant stem cell number, thus improving transplanting both peripheral stem cells and umbilical cord ones. PMID- 10838475 TI - Is there a future for adsorption techniques in sepsis? AB - During sepsis toxins released from, e.g., bacteria induce reactions of various cascade systems that may cause progression of the patient into septic shock, disseminated intravascular coagulation, multiorgan dysfunction syndrome and subsequent death. The use of conventional treatments using antibiotics, fluid substitution, inotropic drugs, respiration aid and dialysis is not enough to reverse the serious prognosis. The addition of various other drugs such as antibodies against various cytokines and cytokine receptors, pentoxiphylline, immunoglobulins or high doses of steroids is usually without benefit for the prognosis of the patient. Another possibility to reduce the extent of toxins and other harmful compounds in the circulation is the use of apheresis (removal by technical devices). This can be done either in a nonselective way (plasma exchange, plasmapheresis) or more selectively using various adsorbers such as polymyxin B. The survival in studies varies between 50 and 80%. Besides the use of nonselective apheresis, the development of various selective adsorption techniques may be one approach to improve survival of these severely ill patients. PMID- 10838476 TI - Extracorporeal removal of circulating immune complexes: from non-selective to patient-specific. AB - The classical immune complex-mediated disease, termed serum sickness, developed a short time after the injection of horse anti-tetanus toxin. Antibodies against circulating horse plasma proteins lead to the formation of immune complexes within the blood circulation (CIC). The inflammatory response, including systemic complement activation and vasculitis, seriously affected the function of all organs, including the most susceptible kidney. Meanwhile CIC have been detected in almost every systemic disease, including autoimmune disorders and also cancer and infections. This brief review will focus on the rationale and the equipment for extracorporeal elimination of CIC. PMID- 10838477 TI - Neural connections of the anterior hypothalamus and agonistic behavior in golden hamsters. AB - In male golden hamsters, offensive aggression is regulated by an interaction between arginine-vasopressin and serotonin at the level of the anterior hypothalamus. The present studies were conducted to study a neural network underlying this interaction. The connections of the anterior hypothalamus were examined by retrograde and anterograde tracing in adult male hamsters. Several limbic areas were found to contain both types of tracing suggesting reciprocal connections with the anterior hypothalamus. Their functional significance relating to the consummation of aggression was tested by comparing neuronal activity (examined through quantification of c-Fos-immunolabeling) in two groups of animals. Experimental animals were sacrificed after attacking an intruder. Control animals were sacrificed after exposure to a woodblock carrying the odor of an intruder that elicited behaviors related to offensive aggression without its consummation. An increased density of Fos-immunoreactivity was found in experimental animals within the medial amygdaloid nucleus, ventrolateral hypothalamus, bed nucleus of the stria terminalis and dorsolateral part of the midbrain central gray. These data suggest that these areas are integrated in a neural network centered on the anterior hypothalamus and involved in the consummation of offensive aggression. Finally, c-Fos-immunoreactivity was combined with labeling of serotonin and vasopressin neurons to identify sub populations particularly associated with offensive aggression. Vasopressin neurons in the nucleus circularis and medial division of the supraoptic nucleus showed increased neuronal activity in the fighters, supporting their role in the control of offensive aggression. PMID- 10838478 TI - Hypothalamic arginine vasotocin mRNA abundance variation across sexes and with sex change in a coral reef fish. AB - Gonadal hormones are important mediators of sexual and aggressive behavior in vertebrates. Recent evidence suggests that the peptide hormones arginine vasotocin (AVT) and its mammalian homologue arginine vasopressin (AVP) often critically mediate these gonadal hormone effects on behavior and have direct influences on behavioral variation. Behavioral differences between sexes, across reproductive states, and even among closely related species are correlated with differences in central AVT/AVP systems in many species. We report differences in hypothalamic AVT mRNA levels between distinct alternate male phenotypes and with female-to-male sex change in the bluehead wrasse (Thalassoma bifasciatum), a teleost fish. The aggressively dominant and strongly courting male phenotype has greater numbers of AVT mRNA producing cells in the magnocellular preoptic area of the hypothalamus than females. Levels of AVT mRNA within these cells in dominant males are also approximately three times female levels whereas the non-aggressive male phenotype has AVT mRNA levels approximately twice female levels. Behavioral sex change is very rapid in this species and is not dependent on the presence of gonads. Conversely, rapid increases in sexual and aggressive behavior during sex change are closely paralleled by approximate fourfold increases in hypothalamic AVT-mRNA levels. The behavioral plasticity shown by bluehead wrasses in response to social environment might be mediated in part by a neuropeptide, AVT, with changes in the gonads and gonadal hormones as the result rather than the cause of behavioral dominance. PMID- 10838479 TI - Neuromuscular correlates to the evolution of flapping flight in birds. AB - The neuromotor pattern (i.e. the onset/offset of muscle contraction within the locomotor cycle) is conserved for some homologous muscles of the tetrapod shoulder but not others in the transition from terrestrial locomotion to flight. Here we test for three shoulder muscles of the European starling (Sturnus vulgaris) to determine whether retention of, or deviation from, a conserved neuromotor pattern can be predicted on the basis of the location of the muscle's motor nucleus within the motor column and the histochemical profile of its constituent muscle fibers. The M. supracoracoideus, the major wing elevator, illustrates a neuromotor pattern that has shifted in its timing within the limb movement cycle. Of the two heads of the triceps, the electrical activity pattern of M. humerotriceps is conserved during the transition, whereas that of the M. scapulotriceps is not. We reacted serial sections of each muscle for myosin adenosine triphosphotase (ATPase), nicotinamide adenine dinucleotide diaphorase (NADH-D), and alpha-glycerophosphate dehydrogenase (alpha-GPD) to characterize all muscles into two fiber types: fast glycolytic (FG) and fast oxidative glycolytic (FOG). We used retrograde axonal tracers to determine the longitudinal distribution and topographical organization of the motoneurons within the motor column in the spinal cord. The histochemical profile of each muscle studied is unique and is statistically different from its homologue in non-avian tetrapods. Compared to non-avian tetrapods, the spatial location of the motor nucleus of the supracoracoideus is conserved. The topology of the two heads of the triceps is fundamentally conserved relative to the other test muscles, but relative to one another there is some spatial segregation which might reflect their respective functional specializations. These data indicate that an evolutionary change in neuromotor pattern can occur without a corresponding topological reorganization of a muscle's motor nucleus within the motor column. Nor can the histochemical profile of homologous muscles be used to predict their neuromotor pattern in the transition from terrestrial locomotion to flight. These findings suggest that evolutionary change in neuromotor outflow relates to altered synaptic input from supraspinal or segmental sources or by alteration of factors intrinsic to individual motoneurons. PMID- 10838480 TI - Phyletic distribution of crypt-type olfactory receptor neurons in fishes. AB - The olfactory epithelium of teleost fishes contains ciliated and microvillous olfactory receptor neurons intermingled with supporting cells. Recently the crypt cell, a third type of olfactory receptor neuron (ORN), was described for two ostariophysans. This type of ORN bears apical microvilli as well as occult cilia extending into a crypt at the apex of the cell. The present study used scanning and transmission electron-microscopic methods to examine how widespread this cell type is in other groups of fish. We investigated the olfactory epithelia of 18 species, freshwater and marine, including various actinopterygian fish as well as 2 species of lungfishes belonging to the sarcopterygians. Crypt cells were detected in 13 species of actinopterygian fish, but in none of the sarcopterygian lungfishes. Crypt cells are present in basic as well as in highly derived actinopterygians. We conclude that crypt cells are a common feature of actinopterygian fish. PMID- 10838481 TI - Historical roots of intolerance in Austrian society. PMID- 10838482 TI - '... As the universities in Austria were more pillars of our movement than those in the old provinces of the Reich'. The University of Vienna from Nazification to de-Nazification. PMID- 10838483 TI - Nazi child 'euthanasia' in Vienna and the scientific exploitation of its victims before and after 1945. PMID- 10838484 TI - The medical profession in Vienna and the Nazi regime. PMID- 10838485 TI - Recollections of an errand boy. PMID- 10838486 TI - Address at the unveiling of the memorial plaque in the courtyard of the University of Vienna. PMID- 10838487 TI - Systemic mastocytosis. AB - Mastocytosis is a disease characterized by an abnormal increase in mast cells. Rare in occurrence, protean in its manifestations, it is a disease which is very seldom thought of and hence, possibly even overlooked. The last few decades have witnessed an upsurge in the understanding of the physiology and pathobiology of mast cells. Better means for diagnosis and follow-up have become available. Once a diagnosis is established histamine antagonists remain the mainstay of treatment. Most patients live and die with the disease rather than of it. The objective of this review is to discuss this entity with a special focus on diagnosis and treatment. PMID- 10838488 TI - Are constipated children different from constipated adults? AB - Constipation represents a diagnostic category of high prevalence both in children and adults, but with low disability and hospitalization rates. This review is written to be a relatively practical guide to physicians providing information for the evaluation and management of one of the most common chronic digestive disorders. In this article we emphasize the differences between constipation in children and adults. The approach to childhood constipation needs to focus on the behavioral nature of this disorder. Diagnostic techniques often used in constipated adults may not provide useful data when applied to children. Treatment of childhood constipation requires a team effort involving the physician, parents and child. PMID- 10838489 TI - A report of a gastrinoma localized preoperatively by endoscopic ultrasound only and a review of the approach to imaging in Zollinger-Ellison syndrome. AB - The addition of somatostatin receptor scintigraphy and endoscopic ultrasound to the preoperative assessment of patients with Zollinger-Ellison syndrome has improved the ability to localize gastrinomas. We report a patient with Zollinger Ellison syndrome with a gastrinoma localized preoperatively by endoscopic ultrasound only. We review the literature regarding the sensitivity of somatostatin receptor scintigraphy and endoscopic ultrasound and discuss the approach to imaging in Zollinger-Ellison syndrome. PMID- 10838490 TI - Clinical image. Unusual endoscopic appearance of a bleeding duodenal ulcer. PMID- 10838491 TI - Sequential trimetrexate, 5-fluorouracil and folinic acid are effective and well tolerated in metastatic colorectal carcinoma. The phase II study group of the AIO. AB - Trimetrexate (TMTX) is a new antifolate which avoids competition for cellular uptake with folinic acid (FA). A regimen of sequential TMTX, FA and 5 fluorouracil (5-FU) has shown efficacy in patients with colorectal cancer. Therefore, we treated 34 previously untreated patients with metastatic colorectal cancer with a weekly chemotherapy regimen consisting of 110 mg/m(2) of TMTX intravenously, then 24 h later 200 mg/m(2) of FA (i.v.) and 500 mg/m(2) of 5-FU (i. v.). Thereafter, 7 doses of oral FA (15 mg) were given at 6-hourly intervals. A treatment cycle consisted of 6 weeks of treatment, then 2 weeks of rest. All patients were treated as outpatients unless complications arose. Thirty-three patients were assessable for tumor response, and all 34 patients were assessable for toxicity. Twelve patients (36%; 95% confidence interval: 25-49%) achieved a partial response. The median duration of response was 8.5 months, and median survival was 14 months. The most common toxicity was diarrhea of grade 3/4, observed in 22% of treatment cycles; this decreased to 8% with early loperamide treatment. Hematologic toxicity was mild. The sequential administration of TMTX, FA and 5-FU is an active regimen in the first-line treatment of metastatic colorectal cancer and warrants further studies. PMID- 10838492 TI - Trends in the clinical characteristics of patients with papillary thyroid carcinoma in Taiwan. AB - The incidence of thyroid cancer is influenced by many factors including socioeconomic status. As economic conditions have improved in Taiwan, the increased frequency of medical examinations in the general population has led to earlier diagnosis of this indolent malignancy. The purpose of this retrospective study was to compare the clinical characteristics of cases of papillary thyroid cancer diagnosed over a 6-year period from 1993 to 1998 with those patients diagnosed from 1977 to 1992 at a single medical center. Of the 1,485 pathologically verified cases of thyroid cancer from 1977 to 1998, 1, 093 had papillary thyroid carcinoma. The mean age of these patients was 40.4 +/- 14.6 years. In order to identify trends in the characteristics of patients with thyroid cancer, patients were divided into those diagnosed before and those diagnosed after 1993. Patients diagnosed in these two time periods were also categorized into disease-free or non-disease-free groups depending on their status at the end of 1998. Actuarial survival rates were calculated using the Kaplan-Meier method. Multivariate analysis was performed to assess the independent effect of these variables using the Cox model. By December 1998, 61 (5.6%) of the 1,093 patients with papillary thyroid carcinoma had died. Among them, only 35 (3.2%) patients had died of thyroid cancer. The 5-year Greenwood survival probabilities for the groups diagnosed before and after 1993 were 0. 9412 and 0.9817, respectively. The patients diagnosed after 1993 received more aggressive surgical procedures, had smaller tumor size, lower postoperative thyroglobulin levels, less advanced clinical stage at the time of diagnosis, showed more disease-free survival, and a lower mortality rate. In conclusion, the results of this study show that patients with a diagnosis of papillary thyroid cancer after 1993 had a smaller tumor size and a better prognosis than those diagnosed before 1993. This finding emphasizes the importance of early detection in thyroid cancer. PMID- 10838493 TI - Anthracycline-based chemotherapy of primary non-Hodgkin's lymphoma of the testis: the hellenic cooperative oncology group experience. AB - Testicular non-Hodgkin's lymphoma is an uncommon disease and its outcome following chemotherapy and/or radiotherapy has been variable. A retrospective analysis was performed on 26 patients with primary testicular lymphoma treated predominantly with anthracycline-based chemotherapy between 1984 and 1999. The patients' median age was 60 years (range 19-82 years) with 17 (65.4%) patients being older than 60 years. Four (15.4%) patients had constitutional B symptoms. There were 11 (42.3%) patients with high grade lymphoma, 12 (46.2%) with intermediate grade, 1 (3.8%) with low grade and 2 (7.7%) were not classified. According to the Ann-Anbor staging system, 18 patients (69.2%) had early (stage I/II) and 8 (30.8%) advanced (stage III/IV) disease. Chemotherapy was administered to 24 patients including 22 patients who received anthracycline based chemotherapy. Two stage IEA patients were treated with orchidectomy and adjuvant radiotherapy to the regional lymph nodes without systemic chemotherapy. Chemotherapy alone resulted in a complete remission (CR) in 14 (58.3%) of 24 patients and partial remission in 1 (4.2%), amounting to an overall response rate (RR) of 62.5%. Of the 5 stage I patients who had chemotherapy on an adjuvant basis, 4 (80%) had CR/no evidence of disease. Of the 11 stage II patients, 8 (72.7%) achieved CR and 1 (9.1%) PR (overall RR of 81.8%). CR was obtained in 2 (25%) of 8 stage III/IV patients. Both patients remain disease free for 26 and 65 months. Excluding the 5 stage I patients, chemotherapy resulted in a CR in 10/19 (52.6%) patients and a PR in 1/19 (5.2%), inducing an overall RR of 57.8%. The mean duration of response was 75 months (range 8-145.5+ months). After a median follow-up of 87 months (range 0.13-145.5+ months) the median survival time was 31 months (range 0.13-145.5+ months) and the median time to progression (TTP) 17 months (range 0.13-145.5+ months). The median TTP was significantly higher in early disease compared to that of advanced disease (52 vs. 3 months, p = 0.02). Of the 3 patients who relapsed following disease-free status, CNS involvement occurred in 2 stage II patients and contralateral testis involvement in 1 stage IEA, respectively. The latter remained disease free for 2 years following orchidectomy alone. The other 2 patients who relapsed did not respond to salvage chemotherapy and died. There was no significant relationship between the values of LDH and beta(2)-microglobulin with the outcome except for ESR which was significantly related with the CR (p = 0.005) or RR (p = 0.005). In conclusion, patients with primary testicular lymphoma have a poor outcome, despite the treatment with anthracycline-containing regimens. Treatment with anthracycline based chemotherapy is recommended in patients at early stages. In advanced disease, more intensive or investigational regimens should be considered. Because the relapse rate in the CNS and contralateral testis is quite high in most studies, prophylactic CNS treatment and radiotherapy to the other testis should be included in the management of testicular lymphoma. PMID- 10838494 TI - Evaluation of the therapeutic effect of transcatheter arterial embolization for hepatocellular carcinoma. AB - Transcatheter arterial embolization (TAE) has been widely performed for patients with hepatocellular carcinoma (HCC). However, the method of evaluating the therapeutic effect of TAE has not been established. We examined the rate of necrotic area to whole tumor (TN) by CT, the tumor regression rate (TR) and the reduction rate in serum alpha-fetoprotein (AFP) levels in patients with HCC who received hepatic resection within 3 months after TAE. In the evaluation of TN, the lipiodol accumulation in tumor was regarded as being necrotic. Rates of necrotic area, which were also examined pathologically (PN) in resected tumors, were compared with TN, TR and AFP reduction rates, respectively. Eighty-eight patients were enrolled in this study, and there was a significant positive correlation between TN and PN (r = 0.80, p < 0.001). Although TR significantly correlated to PN (p = 0.001), the correlation coefficient between them was low (r = 0.34). The correlation coefficients between AFP reduction rate and PN was 0.76 (p < 0.001) in 26 patients (30%) with an AFP level >/=200 ng/ml before TAE. The evaluation method using lipiodol accumulation in CT is the most useful for assessing the therapeutic effect of TAE, particularly when a sufficiently long interval exists between TAE and the evaluation, because of the highest correlation coefficient between TN and PN, and the availability of TN for all patients. The reduction rate in serum AFP levels was also useful in patients with AFP levels >200 ng/ml before treatment. PMID- 10838495 TI - Primary small cell neuroendocrine carcinoma of the vagina. Marked effect of combination chemotherapy: a case report. AB - Primary small cell neuroendocrine carcinoma of the vagina is extremely rare and very aggressive. Nineteen patients with this cancer have been reported in the English-language literature to date. We report a 51-year-old woman with this cancer. Punch biopsy specimens of the vaginal tumor were studied. Electron microscopy demonstrated neuroendocrine granules in the cytoplasm. Tumor cells were strongly stained for cytokeratin, neuron-specific enolase, chromogranin A and serotonin. These findings were consistent with small cell neuroendocrine carcinoma. The patient was treated with combination chemotherapy. The cancer mass completely disappeared after five cycles of the therapy. The patient is still alive without metastasis 41 months after diagnosis. This is the first report of such a remarkable effect of combination chemotherapy on small cell neuroendocrine carcinoma of the vagina. PMID- 10838496 TI - Microsatellite instability in thyroid papillary carcinoma and multinodular hyperplasia. AB - Microsatellite instability (MSI) is a molecular landmark of mutations in DNA mismatch repair genes. The impaired efficiency of DNA repair mechanisms promotes carcinogenesis as well as contributes to tumour progression. Until now, only few studies on MSI in thyroid tumours have been published. Therefore, the aim of the present study was to investigate MSI as a possible characteristic feature of thyroid tumours. The analysis of 12 thyroid papillary carcinomas and 17 multinodular hyperplasias at 13 microsatellite loci showed MSI and loss of heterozygosity (LOH) in both types of lesion, with more alterations noted in the papillary thyroid carcinomas (in 65%) than in multinodular hyperplasia (in 35%). In carcinomas, LOH occurred more frequently than MSI, while in multinodular hyperplasia the LOH/MSI ratio is almost equal. PMID- 10838497 TI - Relative expression of the RET9 and RET51 isoforms in human pheochromocytomas. AB - Activating mutations of the RET oncogene cause the inheritance of multiple endocrine neoplasia type 2 (MEN2). The RET pre-mRNA is spliced into several transcripts coding for multiple isoforms, including Ret9 and Ret51. When harboring activating mutations in the cytoplasmic region, the Ret51 protein displays a higher in vitro transforming efficiency as compared to the corresponding Ret9 isoform. We investigated whether a more transforming isoform was preferentially expressed in MEN2 tumors as compared to normal tissues or sporadic pheochromocytomas. By quantitative RNases protection assays, we measured the absolute abundance of the 3' splice variants in pheochromocytomas and in normal tissues. The proportion of RET51 transcripts was highly dispersed between tumors and normal tissues. In familial tumors the proportion of RET51 transcripts was significantly larger (48.1%) than in sporadic tumors (36.75%). This result suggests that the preferential expression of the Ret51 protein isoform, even though moderate, is a growth advantage for MEN2 tumors. PMID- 10838499 TI - A pilot trial of suramin in metastatic breast cancer to assess antiangiogenic activity in individual patients. AB - Suramin is a polysulfonated naphthylurea with multiple potential mechanisms of action against tumors, including the ability to bind growth factors known to promote tumor angiogenesis. Using an established fixed dosing scheme for the administration of suramin in patients, a pilot study was conducted in patients with progressive, metastatic breast cancer. The primary objective of this trial is to define the effect of suramin on the angiogenic activity in individual patients using in vitro laboratory assays. The secondary objective was to assess the antitumor effect of suramin in a population of metastatic breast cancer patients. No objective tumor responses were observed in any of the 9 patients who received treatment with suramin, however 1 patient did maintain stable disease status. The strength of angiogenic activity present in patient samples was assessed by testing patient plasma in the capillary endothelial cell migration assay. Angiogenic activity followed over time was lowest in patients with the highest suramin concentrations and highest in patients with the lowest suramin concentrations. We conclude that it is feasible to continually monitor the activity of antiangiogenic agents in individual patients without relying on clinical tumor response. PMID- 10838498 TI - RT-PCR suggests human skeletal muscle origin of alveolar soft-part sarcoma. AB - In 1952, Christopherson et al. proposed that alveolar soft-part sarcoma (ASPS) was a distinct entity with unique clinical and pathological features. Since their report, its histogenesis has not been determined. In order to clarify the histogenesis of ASPS, a study using reverse transcription polymerase chain reaction using cDNAs from MyoD1 and myogenin, and the actin filament from human skeletal-muscle-related mRNAs has been performed in 5 cases of ASPS. The expression of MyoD1 and myogenin was determined in 5 and 2 cases of the 5 cases, respectively. Moreover, expression of the many mRNAs from the actin filament of skeletal muscle was also found in ASPS. According to these findings, it is now postulated that ASPS is of skeletal muscle origin. PMID- 10838500 TI - Clinical significance of expression of estrogen receptor alpha and beta mRNAs in ovarian cancers. AB - Novel human estrogen receptor (ER)-beta was identified in cDNA libraries from human testis. ER-beta specifically expresses in testis, ovary, thymus, spleen, osteoblasts and fetus. ER-beta might not conserve the same physiological functions as does ER-alpha. Therefore, the clinical significance of the expression of ER-alpha and ER-beta mRNAs in ovarian cancers was investigated. The percentage of ER-beta mRNA to ER-alpha mRNA ranged from 1.5 to 10% in normal ovaries. On the other hand, the ratios of ER-beta mRNA to ER-alpha mRNA were in a wide range in ovarian cancers. There was no significant difference in the ratios among ovarian cancers classified according to histological types or clinical stages. In a 48-month survival rate, the patient prognosis in ovarian cancers with a low or high ratio of ER-beta mRNA to ER-alpha mRNA (<1.5 or >10% of ER beta mRNA to ER-alpha mRNA) was significantly worse than that in ovarian cancers with a medium ratio (>==1.5 to <==10% of ER-beta mRNA to ER-alpha mRNA). In conclusion, the intact synchronized expression of ER-beta mRNA interacting with ER-alpha mRNA might be damaged in some ovarian cancers, which might lead to poor patient prognosis. PMID- 10838501 TI - Aminopeptidase A expression in cervical neoplasia and its relationship to neoplastic transformation and progression. AB - Aminopeptidase A (AP-A) is a cell surface metallopeptidase which specifically cleaves the amino-terminal acidic residue from peptide substrates such as angiotensin II. AP-A is identical to the differentiation-related antigen, murine BP-1 or human kidney gp160, and is involved in regulating cell differentiation and/or neoplastic transformation of certain normal and transformed cells. We examined expression of AP-A in premalignant and malignant lesions of the uterine cervix, and investigated whether its expression was related to disease progression and neoplastic transformation. Formalin-fixed, paraffin-embedded tissue sections including 14 cervical intraepithelial neoplasms (CIN) and 23 invasive squamous cell carcinomas (SCC) were immunohistochemically evaluated. AP A was localized in the basal cell layer in normal squamous epithelium. In CIN, AP A expression was found on dysplastic cells, and increased with the severity of the precancerous lesions. In invasive cancer, 18 of 19 non-keratinizing-type SCCs and none of 4 keratinizing-type SCCs expressed AP-A. In addition, AP-A immunoreactivity was significantly correlated with proliferating cell nuclear antigen expression in both CIN and SCC cases. Furthermore, angiotensin II type 1 receptor was present in all AP-A-positive SCCs. These results indicate that AP-A is upregulated as the lesion progresses toward carcinoma in the cervical epithelium, and suggest that AP-A may play a regulatory role in neoplastic transformation and disease progression in cervical neoplasms and may serve as a potential tumor marker during cervical neoplasia development. PMID- 10838502 TI - Pediatric neurosurgery: pride and prejudice. AB - Pediatric neurosurgery now exists as a member of the family of neurosurgery with its own training programs, process of accreditation, national and international conferences and scientific journals. The relentless expansion of science relevant to the practice of neurosurgery and the changing patterns of neurosurgical practice have driven and continue to drive the juggernaut of evolutionary process which sometimes necessitates the birth of new specialties of practice. The history and the development of neurosurgery as they relate to children are presented. There is no more reason to think that the established specialty of pediatric neurosurgery or the patients under the care of pediatric neurosurgeons would benefit from the collapsing of pediatric neurosurgery back into the general neurosurgical fold than to think that all of neurosurgery, and hence all patients cared for by neurosurgeons, would benefit from the return of organized neurosurgery to its general surgical parent. Just as mankind benefits from the steady advancement of all aspects of neurosurgery, children benefit from the existence and steady advancement of pediatric neurosurgery. PMID- 10838503 TI - Pump-regulated cerebrospinal fluid drainage. AB - The drainage of cerebrospinal fluid (CSF) from the lumbar subarachnoid space is an effective technique for the treatment of CSF fistula and control of intracranial pressure in children and adults. The use of the lumbar drain poses unique challenges, however, in the pediatric population. We present a safe and effective method of pump-controlled lumbar subarachnoid drainage. This technique allows accurate titration of CSF removal while providing a closed system which is not sensitive to position changes or patient activity. Four case histories are reviewed. PMID- 10838504 TI - Ectopic choroid plexus within a juvenile arachnoid cyst of the cerebellopontine angle: cause of cyst formation or reason of cyst growth. AB - The unusual and rare case of a 6-year-old boy is reported who presented with an arachnoid cyst located in the cerebellopontine angle incorporating an ectopic piece of choroid plexus tissue. A microneurosurgical cyst wall resection was performed and the plexus tissue identified and removed. The rare occurrence of ectopic choroid plexus tissue within cysts of the CNS is discussed. PMID- 10838505 TI - Virtual endoscopy for planning endoscopic third ventriculostomy procedures. AB - Virtual endoscopy was used to plan 10 endoscopic third ventriculostomy procedures in 9 patients suffering from occlusive hydrocephalus due to idiopathic aqueductal stenosis and tumors. The patients were 4 children (4-14 years, mean age: 9 years) and 5 adults (21-38 years, mean age: 29 years). The aim of the study was to preoperatively evaluate the individual intraventricular and vascular endoscopic anatomy at the floor of the third ventricle based on virtual endoscopic images. The virtual views were correlated with the real endoscopic images, intraoperatively obtained by a standard ventriculoscope (Wolf, Knittlingen, Germany) during endoscopic third ventriculostomy procedures. Contrast-medium enhanced MR images (3D-MPRAGE; Siemens, Germany) were semiautomatically segmented with a surface-rendering technique ('Navigator' software; General Electric Medical, Buc, France) to produce the virtual endoluminal views. The virtual endoscopic images were comparable with the real intraoperative endoscopic view in 8 of 9 patients. Virtual endoscopy can display the position of the basilar artery, the posterior cerebral arteries and the posterior communicating arteries in their relationship to the mammillary bodies and the clivus. Preoperative virtual endoscopy planning can intraoperatively assist the neurosurgeon to find a safe location for third ventriculostomy. PMID- 10838506 TI - Terminal Myelocystocele:an unusual presentation. AB - Terminal myelocystocele is an unusual form of occult spinal dysraphism. It consists of a cystic dilatation of a low-lying terminal cord herniated posteriorly through a skin-covered lumbosacral spina bifida. An arachnoid-lined meningocele, continuous with the spinal subarachnoid space, is traversed by the hydromyelic cord. Clinically, this presents with a skin-covered lumbosacral mass, but often no neurological deficit is present. We present a case of terminal myelocystocele in a child born without deficit and without an obvious back mass. Diagnosis was delayed until sphincter disturbance and lower limb inequalities developed. We discuss the presentation, imaging and operative findings in this case. PMID- 10838507 TI - Long-term outcome of terminal myelocystocele patients. AB - INTRODUCTION: A terminal myelocystocele, a closed form of a neural tube defect (NTD), can present as a large, fully epithelialized, cystic lumbosacral mass containing fat, cerebrospinal fluid (CSF) and neural tissue. The spinal cord terminates at a neural placode wherein the central canal opens into a CSF-filled cavity that is distinct from fluid in the subarachnoid space surrounding the spinal cord. This form of NTD, in our experience, was only associated with major caudal cell mass abnormalities, as these patients often have maldevelopment of the lower spine, pelvis, genitalia, bowel, bladder, kidney and the abdominal wall. This study will describe the clinical manifestations, surgical management and long-term outcome of our terminal myelocystocele patients. METHODS: To characterize this rare entity, a 13-year retrospective review was undertaken at our institution. RESULTS: Nine patients with terminal myelocystoceles were identified. In all cases, there were multiple congenital defects including cloacal exstrophy, imperforate anus, omphalocele, pelvic deformity, equinovarus or renal abnormality. Only 1 out of 9 patients has required a shunt for hydrocephalus. The main goal of neurosurgical intervention was to reduce the size of the mass, which can slowly enlarge over time. The spinal cord was also untethered, although these patients have no chance of bowel or bladder control. With a mean follow-up of 63 months, all patients remained neurologically stable. Impairment of lower extremity function is usually severe. However, some patients were ambulatory with the aid of a walker or orthotic device. All patients required a prolonged hospital stay as well as multiple operations prior to initial discharge. CONCLUSION: Experienced, multispecialty care is needed to optimize the long-term outcome of these complex patients. PMID- 10838508 TI - Stroke associated with marijuana abuse. AB - We present the case of a 15-year-old with a cerebellar infarct that involved multiple arterial territories. It was temporally related to, and probably caused by, heavy marijuana use. While the mechanism of marijuana-associated stroke is unclear, the drug is known to cause hypotension and to impair peripheral vasomotor reflexes. We suspect that the child had diminished cerebral autoregulatory capacity and developed the stroke during a period of hypotension. PMID- 10838509 TI - Craniocervical neurenteric cyst without associated abnormalities. AB - Spinal neurenteric (NE) cyst is an uncommon congenital cyst and frequently found in the cervical region. The clinical symptoms associated with this entity depend on the site of the lesion and are not typical for all such cysts. A definitive diagnosis can only be made by biopsy and histological examination. MRI can confirm these cystic masses and is the method of choice for their imaging investigation. They are often connected by a fibrous tract, fistula or cleft to structures derived from the primitive gut in the thoracic or abdominal cavities and are commonly associated with anterior spina bifida or other vertebral anomalies. We report a case of craniocervical NE cyst without associated abnormalities and discuss the implications for clinical diagnosis and management by a thorough review of the literature. PMID- 10838510 TI - A 14-year-Old young woman with a five-week history of back pain. PMID- 10838511 TI - An unusual split cord malformation. PMID- 10838512 TI - Posterior fossa craniotomy: an alternative to craniectomy: an alternative to craniectomy. PMID- 10838513 TI - Effects of handling and storage of blood on the stability of hepatitis C virus RNA: implications for NAT testing in transfusion practice. AB - BACKGROUND AND OBJECTIVES: To determine the stability of hepatitis C virus (HCV) RNA during transport and storage of blood samples from donors, prior to screening for HCV by nucleic acid amplification technology. MATERIALS AND METHODS: Various blood and plasma sample types were stored for up to 120 h at different temperatures and the HCV RNA level was measured using an in house quantitative reverse transcription-polymerase chain reaction. RESULTS: No decline in HCV RNA level was observed after 72 h of storage of whole blood at 4 degrees C in EDTA tubes (Greiner) and Plasma Preparation Tubes (PPT; Becton Dickinson), while insignificant declines of 0.2 log10 and 0. 25 log10 occurred at 25 degrees C after 72 h in the EDTA tubes and PPT tubes, respectively. When whole blood was stored with mixed anticoagulants CPDA-1 and EDTA for up to 120 h, no decline in HCV RNA level was observed at 4 degrees C and 25 degrees C, while a significant decline of 0.37 log10 occurred at 37 degrees C after 120 h. The temperature during transportation was investigated with a 12-hour period at 25 degrees C and 37 degrees C before storage at 4 degrees C for 108 h. Neither temperature resulted in any loss of HCV RNA in comparison with 120 h of storage at 4 degrees C. CONCLUSION: Whole blood anticoagulated with EDTA or CPDA-1/EDTA may be stored at up to 25 degrees C (room temperature) for up to 5 days without any significant loss in plasma HCV RNA level. PMID- 10838514 TI - Assessment of liver disease and biochemical and immunological markers in Swedish blood donors with isolated GB virus C/hepatitis G virus viremia. AB - OBJECTIVE: To investigate signs of liver disease, and biochemical and immunological markers in blood donors with isolated GBV-C/HGV viremia. METHODS: Eighteen donors with isolated GBV-C/HGV viremia were followed up 3-5 years after initial identification. Testing for GBV-C/HGV RNA, GBV-C/HGV-E2 antibodies and a range of biochemical and immunological tests was performed. Thirteen donors consented to liver biopsy. RESULTS: Twelve donors remained GBV-C/HGV viremic at follow-up. Five donors had developed E2 antibodies. Liver biopsies revealed mild portal inflammatory lesions in 6/11 individuals with persistent viremia, and steatosis in 10/13 biopsied donors. CONCLUSION: Steatosis and mild portal inflammatory lesions were found in liver biopsies from several blood donors with isolated GBV-C/HGV viremia. PMID- 10838515 TI - Results of viral marker screening of unpaid blood donations and probability of window period donations in 1997. EPFA Working Group on Quality Assurance. AB - BACKGROUND AND OBJECTIVES: To monitor the safety of the blood supply and evaluate the potential benefits of additional measures, the likelihood of virus transmission must be assessed. The European Plasma Fractionation Association and its member organisations have therefore developed a surveillance system to monitor infection markers among unpaid blood and plasma donors. We report and analyse the results of this surveillance for 1997. MATERIALS AND METHODS: Data from the screening of unpaid donations for anti-HIV, anti-HCV and HBsAg during 1997 were collected retrospectively by EPFA member organisations. We identified seroconverters and estimate the probability of window period donations. RESULTS: Data included screening results from 11 million unpaid donations in Europe, the USA and Australia. Prevalence of viral markers varied, with marker rates from repeat donations in Europe and Australia being significantly lower than in the USA. For first-time donations, in contrast, prevalence of HBsAg in the USA was within the ranges of those measured in Europe and Australia. Screening data of about 5 million European and 0.5 million Australian repeat donations were used to identify seroconverters. From the seroconverters that were detected among the European organisations, we estimated that 1 in every 2,323,778 repeat donations (range: 707,090-20,922,520) was made during the window period of anti-HIV screening. One in 620,754 (201,216-2,316,805) and 1 in 398,499 (155,209 to >1,088,511) repeat donations were made during the anti-HCV or HBsAg window period, respectively. Probabilities of window period donations in Australia were within the ranges of those measured in Europe. CONCLUSIONS: The collated surveillance data of 1997 illustrate the high degree of safety in blood and plasma products from unpaid donors. PMID- 10838516 TI - Continuous monitoring of arterial blood gases and pH during intraoperative rapid blood administration using a Paratrend sensor. AB - BACKGROUND AND OBJECTIVES: The aim of this study was to determine the effects of rapid transfusion of packed red cells on the arterial blood gases and acid-base status of the recipient. MATERIALS AND METHODS: We studied 16 patients (mean age 66.3+/-9.9 years) who received rapid transfusion of 632.8+/-287.2 g of packed red cells in CPDA-1, stored before use for a period of 15.2+/-4.4 days. During transfusion, monitoring of pH, PCO2 and PO2 was continuous using an intra arterial multiparameter sensor (Paratrend 7, Biomedical Sensors, UK). RESULTS: The rate of the transfusion was 73.1+/-9.6 g/min and the duration of observation was 35.8+/-12.8 min. Arterial pH decreased from 7.446+/-0.023 to 7.385+/-0.034 (p<0.001) and PCO2 increased from 32.31+/-1.35 to 36.41+/-1.86 mmHg (p<0.001). Delta pH and delta PCO2 showed significant correlation to the weight and the age of the transfused blood (p<0.001 for both dependent variables). The rate of pH change was positively but insignificantly correlated to the rate of the transfusion. Base excess was significantly decreased and end-tidal CO2 (PetCO2) was increased from 25.8+/-2.0 to 28.1+/-2.3 mmHg (p<0.05), significantly correlating to the amount and age of the administered component (p<0.05). PetCO2 was not elevated when PCO2 changes were minimal. Alterations in PO2 were not specific and our clinical impression was that they were related to unmeasured parameters. CONCLUSION: Our findings suggest that the fall in pH and the elevation in PCO2 which occur during rapid transfusion of packed red cells may go undetected or be misinterpreted if the acid-base status of the recipient is not monitored continuously. These alterations are mainly of metabolic character and depend on the amount and age of the transfused component. Our data suggest that arterial sampling is essential during massive transfusions. PMID- 10838517 TI - Preparation of leukocyte-poor platelet concentrates via a short, hard spin of a pool of buffy coats. AB - BACKGROUND AND OBJECTIVES: A new method for the preparation of leukocyte-poor platelet concentrates was developed, based on a short, hard spin of a pool of 5 buffy coats (BCs) combined with automated collection of the platelets. MATERIALS AND METHODS: The characteristics of platelet concentrates (PCs) were studied as a function of the total g force applied to a pool of 5 BCs. Pools of BCs were centrifuged for 1 min with a total g force ranging from about 3,300 to 5,000 gmin (n = 7-9 per applied g force). Deceleration took place without the means of a brake. The total centrifugation time was about 11 min. The platelet-rich plasma (PRP) fraction above the cell layer was separated by an automated component preparation device. RESULTS: A short, hard spin with a total g force of between 3,400 and 4,600 gmin resulted in PCs that contained on average more than 290x10(9) platelets and less than 5x10(6) leukocytes without the use of a leukocyte filter, provided that the transfer of PRP was electronically checked and terminated. The cell concentrations in the PCs are a function of the total g force. Both the platelet and leukocyte levels in the concentrate decreased with an increase in the total g force applied to the pool. CONCLUSION: The preparation of PCs via a short hard, spin of BCs, combined with automated collection of the PRP, may be an alternative method for the preparation of leukocyte-poor PCs. PMID- 10838518 TI - Are all leucodepleted platelet concentrates equivalent? Comparison of Cobe LRS Turbo, Haemonetics MCS+ LD, and filtered pooled buffy-coat-derived platelets. AB - BACKGROUND AND OBJECTIVES: A number of technologies are available for the production of leucocyte-depleted platelet concentrates (PCs). This study compared the characteristics of PCs prepared by three commonly used techniques. MATERIALS AND METHODS: In total, fifteen units of leucocyte-depleted PCs prepared by the Cobe LRS Turbo apheresis system, Haemonetics MCS+ with in-line filter and filtration of PCs derived from pooled buffy coats (BCs) were transferred into the same standard container. Markers to assess status/activation and microvesiculation of platelets as well as platelet injury were measured. RESULTS: pH was well maintained in all types of PCs. The expression of CD62P was higher in Cobe LRS Turbo on day 1 but became equivalent between the three methods on day 5. A significant correlation was found between the expression of CD62P on platelet surface and soluble CD62P in the plasma. The degree of phosphatidylserine (PS) exposure was slightly higher in Cobe LRS Turbo and BC-PCs than Haemonetics MCS+ on day 1. However, on day 5 both apheresis PC values were higher than BC-PCs. A significant correlation was found between PS exposure and microvesiculation. CONCLUSION: Leucodepleted PCs prepared by the three methods were different in terms of storage lesion and microvesiculation. The clinical significance of these findings remains to be investigated. PMID- 10838519 TI - Storage of platelets in additive solutions: effects of phosphate. AB - BACKGROUND AND OBJECTIVES: In a previous study, low adenine nucleotide levels and a reduced rate of glycolysis were found in platelet concentrates (PCs) prepared by apheresis and stored in a platelet additive solution (PAS). Our objective was to investigate whether the use of PAS with or without phosphate can influence platelet metabolism in a similar way. MATERIALS AND METHODS: The in vitro effects of storage in either plasma or a PAS (T-Sol or PAS-III, both containing citrate, acetate and sodium chloride, PAS-III containing also phosphate) of buffy-coat derived pooled platelet concentrates (BC-PCs) and apheresis platelets were investigated. The use of PAS implies inclusion of some plasma (20 or 35%). Paired studies over 7 days included investigation of cell counts, pH, PO2, PCO2, bicarbonate, glucose, lactate, adenine nucleotides, and extracellular adenylate kinase activity as a marker for disintegration of platelets. The expected concentration of phosphate in T-Sol is 0.6-1.8 mmol/l (with CPD plasma) and 0.2 0.6 mmol/l (with ACD plasma), and in PAS-III, 15-25 mmol/l (calculated values). RESULTS: BC-PCs were compared during storage in 35% CPD plasma and 65% PAS (T-Sol or PAS-III) (experiment 1), or alternatively 20% CPD plasma and 80% PAS (T-Sol or PAS-III) (experiment 3). In both studies, PAS-III shows similar and significantly higher rates of glycolysis in terms of consumption of glucose (0.06 vs. 0.04 mmol/day/10(11) platelets) and production of lactate (0.11 vs. 0.07 mmol/day/10(11) platelets) compared with T-Sol. Levels of pH and adenine nucleotides were similar when 35% plasma was used. With only 20% plasma, significantly higher levels of adenine nucleotides were found with PAS-III compared to T-Sol. The storage of apheresis platelets in 35% ACD plasma and 65% PAS (either T-Sol or PAS-III) (experiment 5) gave significantly higher values for PAS-III compared to T-Sol with regard to consumption of glucose (0.08 vs. 0.06 mmol/day/10(11) platelets), production of lactate (0.14 vs. 0.11 mmol/day/10(11) platelets) and adenine nucleotide levels. CONCLUSION: With respect to apheresis PCs stored in media containing ACD plasma, our results suggest that the differences found are related to the concentration of phosphate. The results for BC-PCs stored in media containing CPD plasma suggest that PAS-III is preferable to T-Sol as the PAS at plasma concentrations below 35%. The mechanism behind the phenomena observed with BC-PCs is not known. PMID- 10838520 TI - Flow-cytometric method for the quantitation of the Fc function of intravenous immunoglobulin preparations. AB - OBJECTIVES: We have developed and optimised a new flow-cytometric method for the measurement of the Fc function of intravenous immunoglobulin (IVIg) preparations, which is important in predicting the effector function of immunoglobulin (Ig) in such preparations. MATERIALS AND METHODS: Ig was bound to a monocytic cell line, THP-1 with Fc gammaRI and Fc gammaRII cell surface receptors, and the bound Ig detected by FITC-conjugated F(ab)2 fragment of rabbit anti-human IgG. RESULTS: Validation studies showed that Ig bound to the cell line through the Fc portion. The method detected alterations in Fc function caused by reduction with dithiothreitol or by storage. The method was reproducible (CV<11%) and a limited comparison study showed that it correlated with the European Pharmacopoeia reference method. CONCLUSIONS: This technically simple method is suitable for the quantitation of the Fc function of Ig preparations. PMID- 10838521 TI - D(Va) category phenotype and genotype in Japanese families. AB - BACKGROUND AND OBJECTIVES: The genetic background of the D(Va) category phenotype has been described in two Caucasian individuals. We were interested in the RHD sequence of 7 Japanese D(Va) individuals and their families. MATERIALS AND METHODS: With SSP-PCR we tested exons 4, 5 and 7 of the RHD gene and used restriction enzymes for testing nucleotide associated with the D(Va) phenotype. RESULTS: A single RHD G667 C697 allele was present in 5 individuals with D(Va) category phenotype, and in 2 individuals we found a D-CE-D hybrid gene (exon 5 had been replaced). The D(Va)Ce gene complex was found in all families. CONCLUSION: The changes of the RHD gene described in European D(Va) individuals were also observed in Japanese families. PMID- 10838522 TI - Anti-HEV in blood donors: what is test and what is virus? PMID- 10838523 TI - The detection of alloantibodies against red cells in patients with warm-type autoimmune haemolytic anaemia. PMID- 10838524 TI - Controlled postcardioplegia reperfusion: mechanism for attenuation of reperfusion injury. AB - OBJECTIVE: Controlled reperfusion and secondary cardioplegia are used to minimize reperfusion injury. The mechanisms for their benefit are incompletely defined and may include attenuation of myocyte sodium uptake. METHODS: Pigs had 1 hour of cardioplegic arrest followed by reperfusion with blood (control) or warm cardioplegic solution followed by blood (test). Reperfusion injury in the control and test groups was quantified by measuring changes of intramyocyte ion content with atomic absorption spectrometry and by analyzing electrophysiologic recovery from recordings of reperfusion arrhythmias. RESULTS: Control animals had an increase in intramyocyte sodium content at 5 minutes after initiating reperfusion (+20.2 micromol/g dry weight, P <.04), whereas the test group had an insignificant decrease (-14.0 micromol/g dry weight, P =.33). The first rhythm after initiating reperfusion was more often ventricular fibrillation in the control group (100% vs 50%, P <.02), and the control group required more defibrillations to establish a nonfibrillating rhythm (4.5 +/- 1.2 vs 1.1 +/- 0.3, P <.03). CONCLUSIONS: Controlled reperfusion eliminated the increase in intramyocyte sodium that was observed in the control group at 5 minutes after cardioplegic arrest. This improvement in myocyte ion homeostasis during postcardioplegia reperfusion was associated with fewer reperfusion arrhythmias. These data support the hypothesis that attenuation of myocyte sodium gain during postischemic reperfusion is a mechanism by which controlled reperfusion and secondary cardioplegia are beneficial. PMID- 10838525 TI - Effects of retrograde cardioplegia on myocardial perfusion and energy metabolism in immature porcine myocardium. AB - OBJECTIVES: Retrograde cardioplegia has been widely used for the protection of adult hearts during cardiac operations. Its efficacy to protect immature myocardium is still unclear. This study was designed to assess the effects of retrograde cardioplegia on myocardial perfusion and energy metabolism in immature hearts. METHODS: Piglet hearts were divided into 3 groups. Hearts in group 1 were used to assess myocardial perfusion of retrograde cardioplegia by means of magnetic resonance imaging. Hearts in groups 2 and 3 were used to assess the effects of retrograde cardioplegia on myocardial energy metabolism by use of phosphorus 31 magnetic resonance spectroscopy. RESULTS: Magnetic resonance images showed that perfusion with retrograde cardioplegic solution was heterogeneous. A perfusion defect was noted during retrograde cardioplegia in the right ventricular wall and in a portion of the posterior wall of the left ventricle in 4 of 6 hearts. Phosphorus 31 spectra showed that at the end of 45-minute retrograde cardioplegia, myocardial intracellular pH was 6.83 +/- 0.17 and phosphocreatine was 53.5% +/- 27% of its prearrest value. The adenosine triphosphate level, however, remained normal throughout the retrograde cardioplegia period. Last, the hearts subjected to retrograde cardioplegia or antegrade cardioplegia showed similar and complete metabolic and functional recovery during reperfusion. CONCLUSIONS: Retrograde cardioplegia provides heterogeneous perfusion. Its ability to protect the right ventricular myocardium is poor and varies between individuals. Myocardial perfusion provided by retrograde cardioplegia is slightly less than that needed to sustain normal myocardial energy metabolism under normothermic conditions. PMID- 10838526 TI - Early results of the fontan procedure in one hundred consecutive patients with hypoplastic left heart syndrome. AB - OBJECTIVES: The purpose of this study was to review a large, single institutional experience with the Fontan procedure for patients with hypoplastic left heart syndrome. METHODS: One hundred consecutive patients with "classic" hypoplastic left heart syndrome underwent Fontan palliation between February 1992 and April 1998. Patient demographic, morphologic, and procedural variables were examined and analyzed. In particular, two different surgical techniques were used: technique I (February 1992 to December 1995) employed cardiopulmonary bypass and moderate systemic hypothermia, and technique II (December 1995 to April 1998), profound hypothermia and circulatory arrest. A retrospective review of medical records was performed and variables were examined and analyzed. RESULTS: Hospital survival for the entire cohort was 89% (95% CI 83%-95%). The technique of operation, cardiopulmonary bypass time, and aortic crossclamp time were each strongly associated with survival. Survival for patients treated by technique I was 79% (95% CI 68-91%; n = 48) and for those treated by technique II, 98% (95% CI 94%-100%; n = 52). Cardiopulmonary bypass and crossclamp times were also highly correlated with time to extubation and length of intensive care unit stay. Preoperative pulmonary artery pressure was correlated with survival; preoperative oxygen saturation, right atrial pressure, pulmonary vascular resistance, pulmonary artery size, extent of aortopulmonary artery collaterals, and echocardiographic estimates of ventricular function and tricuspid regurgitation were not correlated with survival. CONCLUSIONS: Our recent experience with Fontan palliation for patients with hypoplastic left heart syndrome suggests that it is attended by low perioperative mortality. The precise operative technique used appears to be an important determinant of outcome, with the duration of cardiopulmonary bypass and crossclamping being particularly significant. PMID- 10838527 TI - Prognostic value of the quantified expression of p185 in non-small cell lung cancer. AB - BACKGROUND: We sought to assess the relationship between tissue concentration of erb -b-2 or neu oncogene-encoded protein (p185(neu)) with overall survival in patients with non-small cell lung cancer. METHODS: Levels of protein p185(neu) were determined in 102 patients with the diagnosis of non-small cell lung cancer. Concentration of p185(neu) protein was determined by using enzyme immunoassay and evaluated by using several variables. The relative prognostic importance of this marker and its influence on other prognostic factors was evaluated by using the Cox regression model. RESULTS: The mean p185(neu) value in these samples was 250 +/- 200 U/mg (95% confidence interval, 210-290). This distinguished two groups within the tumoral population: those with less than 350 U/mg and those with 350 U/mg or greater (80th percentile). Multivariable analysis established an independent prognostic value for protein p185(neu). Patients with p185(neu) values of the 80th percentile or greater had a risk of death that was 2.11-fold (95% confidence interval, 1.10-4.05) that of patients with values of less than 350 U/mg (P =.03), and increases in the neu oncogene of 100 U/mg increased the probability of death by 17% (P =.02; 95% confidence interval, 1.04-1.31). CONCLUSION: This study shows that the p185(neu) expression is an objective and comparable variable for the assessment of phenotypic aggressivity in non-small cell lung cancer, and in the future, it could be included in daily clinical practice. PMID- 10838528 TI - Effect of operative volume on morbidity, mortality, and hospital use after esophagectomy for cancer. AB - OBJECTIVE: We sought to evaluate the effect of operative volume, hospital size, and cancer specialization on morbidity, mortality, and hospital use after esophagectomy for cancer. METHODS: Data derived from the Health Care Utilization Project was used to evaluate all Medicare-reimbursed esophagectomies for treatment of cancer from 1994 to 1996 in 13 national cancer institutions and 88 community hospitals. The complications of care, length of stay, hospital charges, and mortality were assessed according to hospital size (>/=600 beds vs <600 beds), cancer specialization (national cancer institution vs community hospital), and operative volume (esophageal [>/=5 Medicare esophagectomies per year vs <5 Medicare esophagectomies per year] and nonesophageal operations [>/=3333 cases per year vs <3333 cases per year]). RESULTS: Mortality was lower in national cancer institution hospitals (4.2% [confidence interval, 2.0%-6.4%] vs 13.3% [confidence interval, 4.2%-26.2%], P =. 05) and in hospitals performing a large number of esophagectomies (3. 0% [confidence interval, 0.09%-5.1%] vs 12.2% [confidence interval, 4.5%-19.8%], P <.05). Multivariate analysis revealed that the independent risk factor for operative mortality was the volume of esophagectomies performed (odds ratio, 3.97; P =.03) and not the number of nonesophageal operations, hospital size, or cancer specialization. Hospitals performing a large number of esophagectomies also showed a tendency toward decreased complications (55% vs 68%, P =.06), decreased length of stay (14.7 days vs 17.7 days, P =.006), and decreased charges ($39,867 vs $62, 094, P <.005). CONCLUSIONS: These results demonstrate improved outcomes and decreased hospital use in hospitals that perform a large number of esophagectomies and support the concept of tertiary referral centers for such complex oncologic procedures as esophagectomies. PMID- 10838529 TI - The role of computed tomographic scanning in diagnosing mediastinal node involvement in non-small cell lung cancer. AB - OBJECTIVES: The reliability of computed tomographic scanning in evaluating mediastinal node involvement is controversial because of the high false result rate. We attempted to identify significant factors responsible for false-positive and false-negative scans. METHODS: From August 1992 through April 1997, 401 patients with lung cancer who underwent major lung resection and systematic lymph node dissection were enrolled in this study. We retrospectively examined mediastinal node size, tumor location, maximum tumor dimension, the presence or absence of obstructive pneumonia, atelectasis, pulmonary fibrosis, and lymph node calcification on contrast-enhanced computed tomographic scans. We identified clinical and radiologic factors responsible for the false results by using univariate and multivariable analysis. RESULTS: Central tumor location proved to be a significant factor of false-positive scans. Elevated carcinoembryonic antigen level and larger tumor dimension were significant factors of false negative scans. In patients with a peripheral tumor smaller than 40 mm and normal levels of serum carcinoembryonic antigen, sensitivity, specificity, positive predictive value, and negative predictive value were 6%, 93%, 8%, and 90%, respectively. The reliability of computed tomographic scanning in this low-risk subgroup was high in detecting N0-1 disease but low in diagnosing N2 disease. CONCLUSION: It is not possible to accurately diagnose N2 disease by using lymph node size on computed tomographic scanning alone, especially in patients with a central tumor, an elevated serum carcinoembryonic antigen level, or a tumor of 40 mm or larger. A preoperative invasive staging procedure is indicated in these populations and may not be indicated in the population with normal computed tomographic scan results without any of these risk factors. PMID- 10838530 TI - Results of surgical treatment of non-small cell lung cancer: validation of the new postoperative pathologic TNM classification. AB - OBJECTIVE: Prognostic relevance of the current TNM stage grouping for lung cancer is still a matter of debate. METHODS: To validate the new pathologic TNM classification for non-small cell lung cancer, we analyzed the survival data of 586 patients who underwent complete pulmonary resection and pathologic staging at one institution. RESULTS: The current TNM stage grouping well reflected the long term prognostic hierarchy. There was a good distinction between new substages IA and IB (5-year survivals of 66% and 53%, respectively). The subdivision of stage II led to an under-representation of stage IIA (6 patients [1.0%]), and therefore the appropriateness of this modification could not be verified. Five-year survival in the T3 N0 category (30%) was significantly better than that found in the new stage IIIA (15%). No difference was found between T3 N0 and T2 N1, the categories constituting new stage IIB. Within stage IIIA there was a significant survival difference between T3 N2 (6%) and the remaining T and N designations (18%). Moreover, the 5-year survival in the T3 N1 category (35%) was similar to that found in the new stage IIB (27%) and better than in any T N2 tumors (12%). CONCLUSION: Most of our findings confirmed prognostic relevance of the current pTNM stage grouping in patients with resectable non-small cell lung cancer. However, despite recent modifications, there is still a significant heterogeneity that flaws stage IIIA. PMID- 10838531 TI - Factors determining outcome after surgical resection of T3 and T4 lung cancers of the superior sulcus. AB - BACKGROUND: The treatment of superior sulcus lung cancers is evolving and preoperative chemotherapy is increasingly used. To establish a historical benchmark against which new therapies can be assessed, we reviewed our 24-year experience with patients undergoing thoracotomy for lung cancers of the superior sulcus. METHODS: Data were acquired through retrospective chart review. Overall survival was calculated by the method of Kaplan and Meier, and prognostic factors were examined by log rank and Cox proportional hazards modeling. RESULTS: From 1974 to 1998, 225 patients underwent thoracotomy. The patients included 144 men (64%) and 81 women with a median age of 55 years. The majority of patients (55%) received preoperative radiation, but 35% did not have any preoperative treatment. Tumor stages were IIB (T3 N0) in 52%, IIIA in 15%, and IIIB in 27% of patients. Complete resection was achieved in 64% of T3 N0 tumors, 54% of T3 N2 tumors, and 39% of T4 N0 tumors. Operative mortality was 4%. Median survival was 33 months for stage IIB and 12 months for both stages IIIA and IIIB. Actuarial 5-year survivals were 46% for stage IIB, 0% for stage IIIA, and 13% for stage IIIB. By univariate and multivariable analyses, T and N status and complete resection had a significant impact on survival. Locoregional disease was the most common form of relapse. CONCLUSIONS: Our results provide a benchmark against which new treatment regimens can be evaluated. Control of locoregional disease remains the major challenge in treating lung cancers of the superior sulcus. The potential benefit of preoperative chemotherapy or chemoradiotherapy must be assessed by whether it leads to higher rates of complete resection and a lower risk of local relapse. PMID- 10838532 TI - Ewing sarcoma of the rib: results of an intergroup study with analysis of outcome by timing of resection. AB - OBJECTIVE: We sought to establish the outcome and optimal therapeutic sequence for patients with nonmetastatic Ewing sarcoma/primitive neuroectodermal tumor of the chest wall. METHODS: Patients 30 years of age or younger with nonmetastatic Ewing sarcoma/primitive neuroectodermal tumor of the bone were randomly assigned to receive vincristine, doxorubicin, cyclophosphamide, and dactinomycin or those drugs alternating with ifosfamide and etoposide. Local control was obtained with an operation, radiotherapy, or both. RESULTS: Fifty-three (13.4%) of 393 patients had primary tumors of the chest wall (all rib). Event-free survival at 5 years was 57% for the chest wall compared with 61% for other sites (P >.2). Ifosfamide and etoposide improved outcome in the overall group (5-year event-free survival, 68% vs 54%; P =.002), and a similar trend occurred in chest wall lesions (5-year event-free survival, 64% vs 51%). Patients with chest wall lesions had more attempts at initial surgical resection (30%) than those with other primary tumor sites (8%, P <.01). The attempt at initial resection for chest wall lesions did not correlate with size. Initial resections at other sites were restricted to smaller tumors. Initial resection resulted in negative pathologic margins in 6 of 16 patients, whereas the delayed resection resulted in negative margins in 17 of 24 patients (P =.05). Although there was no difference in survival by timing of the operation in rib lesions, a higher percentage of patients with initial surgical resection received radiation than those with resection after initial chemotherapy (P =. 13). CONCLUSIONS: Although rib primary tumors are significantly larger than tumors found in other sites, their outcome is similar. We favor delayed resection whenever possible to minimize the number of patients requiring radiation therapy. PMID- 10838533 TI - Carinal reconstruction with a Y-shaped collagen-conjugated prosthesis. AB - BACKGROUND: Carinal reconstruction by direct suturing is associated with a high mortality because high tension at the anastomosis can lead to tracheobronchial fistula. A new tracheal prosthesis was therefore designed for reconstruction of the tracheal bifurcation and applied for experimental carinal replacement in dogs. METHODS: The main frame of the new prosthesis consists of a Y-shaped Marlex mesh tube (C.R. Bard, Inc, Billerica, Mass) reinforced with polypropylene spirals, to which collagen extracted from porcine skin is chemically conjugated to provide biocompatibility and airtightness. This conjugated collagen is composed of amorphous and sponge collagen layers. The tracheobronchial bifurcation was replaced with the prosthesis in 10 beagle dogs. RESULTS: Eight dogs survived the postoperative period, and 2 dogs died within 4 days after the operation. Bronchoscopic examination revealed that the tracheal prosthesis was covered with smooth whitish tissue and that no stenosis or dehiscence was present in the 8 dogs even 6 months after the operation. Slight mesh exposure was recognized in 1 dog. Histologic examination revealed that approximately one half of the luminal surface was covered with ciliated columnar epithelium or nonciliated squamous epithelium. In the remaining lumen, especially in the middle portion of the prosthesis, connective tissue without epithelium was observed. CONCLUSIONS: These long-term results indicate that our bifurcated tracheal prosthesis can be used for reconstruction of the tracheobronchial bifurcation with long-term safety. PMID- 10838534 TI - Comparison of the effects of fetal cardiomyocyte and skeletal myoblast transplantation on postinfarction left ventricular function. AB - OBJECTIVES: Transplantation of fetal cardiomyocytes improves function of infarcted myocardium but raises availability, immunologic, and ethical issues that justify the investigation of alternate cell types, among which skeletal myoblasts are attractive candidates. METHODS: Myocardial infarction was created in rats by means of coronary artery ligation. One week later, the animals were reoperated on and intramyocardially injected with culture growth medium alone (controls, n = 15), fetal cardiomyocytes (5 x 10(6) cells, n = 11), or neonatal skeletal myoblasts (5 x 10(6) cells, n = 16). The injections consisted of a 150 microL volume and were made in the core of the infarct, and the animals were immunosuppressed. Left ventricular function was assessed by echocardiography immediately before transplantation and 1 month thereafter. Myoblast-transplanted hearts were then immunohistologically processed for the expression of skeletal muscle-specific embryonic myosin heavy chain and cardiac-specific connexin 43. RESULTS: The left ventricular ejection fraction markedly increased in the fetal and myoblast groups from 39.3% +/- 3.9% to 45% +/- 3.4% (P =.086) and from 40.4% +/- 3.6% to 47.3% +/- 4.4% (P =.034), respectively, whereas it decreased in untreated animals from 40.6% +/- 4% to 36.7% +/- 2.7%. Transplanted myoblasts could be identified in all animals by the positive staining for skeletal muscle myosin. Conversely, clusters of connexin 43 were not observed on these skeletal muscle cells. CONCLUSIONS: These results support the hypothesis that skeletal myoblasts are as effective as fetal cardiomyocytes for improving postinfarction left ventricular function. The clinical relevance of these findings is based on the possibility for skeletal myoblasts to be harvested from the patient himself. PMID- 10838536 TI - Commentary PMID- 10838535 TI - Insulin cardioplegia for elective coronary bypass surgery. AB - BACKGROUND: Improved methods of myocardial preservation are required to reduce the morbidity and mortality of coronary bypass surgery for high-risk subgroups. Metabolic stimulation with insulin, glucose solutions, or both has been proposed as a method to preserve the ischemic myocardium. We performed a prospective, double-blind, randomized trial to evaluate the effects of insulin and glucose as cardioplegic additives when used as part of a tepid continuous blood cardioplegic strategy. METHODS: We randomized 56 male patients undergoing elective isolated coronary bypass surgery to 1 of 4 cardioplegic groups containing either 42 or 84 mmol/L glucose with or without 10 IU/L of insulin. Perioperative assessments of myocardial metabolism and left ventricular function were performed. RESULTS: Insulin-enhanced cardioplegia was associated with beneficial effects on both myocardial metabolic and functional recovery after cardioplegic arrest. Insulin's effect was independent of the ambient glucose concentration. CONCLUSIONS: Cardioplegic formulations containing a 42 mmol/L concentration of glucose and a 10 IU/L concentration of insulin provide significant benefit to patients undergoing isolated coronary bypass surgery. The clinical effect of these formulations will need to be assessed in high-risk subgroups of patients, such as those with unstable angina, recent myocardial infarction, or poor left ventricular function. PMID- 10838538 TI - Commentary PMID- 10838537 TI - Aortic valve replacement with the freestyle stentless bioprosthesis with respect to spacial orientation of patient coronary ostia. AB - OBJECTIVE: This study evaluates our results for safety and efficacy of aortic valve replacement using the Freestyle bioprosthesis (Medtronic, Inc, Minneapolis, Minn) with a new modified subcoronary implantation technique. This technique takes into account the spacial orientation of the stentless bioprosthesis in the aortic root with respect to the patient's coronary ostia rather than the native commissures. METHODS: Fifty-two consecutive patients with predominant aortic valve stenosis underwent aortic valve replacement with a Freestyle bioprosthesis by means of the described modified subcoronary technique over a 15-month period. Fifty of them were followed up by means of echocardiography at discharge, 6 months, and 1 year. There were 19 men and 31 women, with a mean age of 76 +/- 7 years (range, 58-87 years). Valve size ranged from 21 to 27 mm. RESULTS: Patients with bicuspid aortic valves had a significantly larger angle between both coronary ostia than patients with tricuspid aortic valves (P =.0001). The peak and mean systolic gradients decreased significantly during the first postoperative year for each valve size (P .1). The cerebral oxygen metabolism fell to 66.0% +/- 22.3% of the level before deep hypothermic circulatory arrest, which was greater than the recovery in the control animals but not significantly so (P =.5). After deep hypothermic circulatory arrest, the renal blood flow fell to 81.0% +/- 29.5% of the value before deep hypothermic circulatory arrest (P =.06). Improvement in renal blood flow in the leukocyte filter group was not significantly greater than the recovery to 70.2% +/- 26.3% in control animals (P =.47). CONCLUSIONS: After a period of deep hypothermic circulatory arrest, there is a significant reduction in cerebral blood flow, cerebral oxygen metabolism, and renal blood flow. Leukocyte depletion with an in-line arterial filter does not appear to significantly improve these findings in the neonatal piglet. PMID- 10838547 TI - Stimulation of neutrophil integrin expression during coronary artery bypass grafting: comparison of crystalloid and blood cardioplegic solutions. AB - OBJECTIVES: This study was designed (1) to evaluate the influence of plasma obtained from patients undergoing coronary artery bypass grafting on L-selectin, CD11b, and CD18 expression on human neutrophils and (2) to determine the influence of the use of crystalloid or blood cardioplegia during bypass grafting on plasma-mediated expression of adhesion molecules on polymorphonuclear neutrophils. PATIENTS AND METHODS: Patients undergoing coronary artery bypass grafting were divided into 2 groups to receive crystalloid or blood cardioplegic solutions. Peripheral vein, radial artery, and coronary sinus blood samples were drawn at aortic crossclamping, aortic crossclamp release, and 30 minutes after reperfusion. Human neutrophils were incubated with patients' plasma, and the expression of CD11b, CD18, and L-selectin was determined with flow cytometry. RESULTS: In patients receiving crystalloid cardioplegic solutions, plasma samples collected from the coronary sinus at aortic clamp release and 30 minutes thereafter induced significantly higher expression of neutrophil CD11b and CD18 than plasma samples obtained from a peripheral vein or artery at the same time points. The expression of L-selectin on polymorphonuclear neutrophils was significantly reduced with plasma obtained 30 minutes after reperfusion as compared with samples collected at aortic crossclamp release. In the group receiving blood cardioplegia, no significant differences in CD11b, CD18, or L selectin expression were found. CONCLUSIONS: (1) Ischemia/reperfusion after coronary artery bypass grafting is associated with the release of factors capable of neutrophil activation from myocardium into the circulating blood. (2) The release of soluble stimuli for neutrophils during bypass grafting may be modified by the cardioplegic solution. PMID- 10838548 TI - A quick assay for monitoring recombinant hirudin during cardiopulmonary bypass in patients with heparin-induced thrombocytopenia type II : adaptation of the ecarin clotting time to the act II device. AB - BACKGROUND: Recombinant hirudin is increasingly advocated as a promising alternative anticoagulation for patients with heparin-induced thrombocytopenia type II during cardiopulmonary bypass. This requires monitoring of the ecarin clotting time. No commercial ecarin clotting time assay is available for clinical use. We adapted the ecarin clotting time to the easy-to-handle ACT II device. METHODS: Three different concentrations of the ecarin reagent (20, 10, 5 U/mL) were investigated as preliminary studies. Standard calibration curves were constructed for concentrations of recombinant hirudin ranging from 0 to 5 microgram/mL. In vivo samples were collected from patients with heparin-induced thrombocytopenia type II who underwent cardiopulmonary bypass, and the values were compared with the values obtained by the chromogenic method. The final concentration for the assay of 5 IU/mL ecarin was further assessed in vitro for reproducibility and the influence of variations in hematocrit, platelet count, and procoagulants. RESULTS: All three concentrations of ecarin revealed linearity to 5 microgram/mL concentrations of recombinant hirudin. The ecarin concentration of 5 U/mL revealed the best correlation (0.87) to the laboratory method, was reproducible over the whole recombinant hirudin range, and was not influenced by the variations in the in vitro setup. CONCLUSIONS: The ACT II/ecarin clotting time with an ecarin concentration of 5 U/mL is a simple and reliable assay for monitoring recombinant hirudin during cardiopulmonary bypass. Use of this assay allows a wider use of recombinant hirudin in patients with heparin-induced thrombocytopenia type II during bypass and thereby may contribute to the safer management of these patients. PMID- 10838549 TI - Aortopulmonary window associated with complete atrioventricular septal defect. PMID- 10838550 TI - Successful local excision and long-term survival for invasive pulmonary aspergillosis during neutropenia after bone marrow transplantation. PMID- 10838551 TI - Tumor implantation after computed tomography-guided biopsy of lung cancer. PMID- 10838552 TI - Combined sleeve lobectomy and Ivor lLwis esophagectomy for synchronous primary carcinoma of the lung and Barrett esophagus. PMID- 10838553 TI - Second operation for upper mediastinal lymphadenopathy after complete resection for cancer of the left lung. PMID- 10838554 TI - Patch closure of the aortic anulus in a recipient of a ventricular assist device. PMID- 10838555 TI - Paraplegia after coronary artery bypass operations: relationship to severe hypertension and vascular disease. PMID- 10838557 TI - Use of intravenous methylene blue for the treatment of refractory hypotension after cardiopulmonary bypass. PMID- 10838558 TI - Notes about axillary cannulation. PMID- 10838559 TI - Is neck or chest anastomosis preferable during esophageal resection? PMID- 10838561 TI - p38 MAP kinases: beyond the stress response. PMID- 10838562 TI - Evolution of a metabolic pathway for degradation of a toxic xenobiotic: the patchwork approach. AB - The pathway for degradation of the xenobiotic pesticide pentachlorophenol in Sphingomonas chlorophenolica probably evolved in the past few decades by the recruitment of enzymes from two other catabolic pathways. The first and third enzymes in the pathway, pentachlorophenol hydroxylase and 2,6 dichlorohydroquinone dioxygenase, may have originated from enzymes in a pathway for degradation of a naturally occurring chlorinated phenol. The second enzyme, a reductive dehalogenase, may have evolved from a maleylacetoacetate isomerase normally involved in degradation of tyrosine. This apparently recently assembled pathway does not function very well: pentachlorophenol hydroxylase is quite slow, and tetrachlorohydroquinone dehalogenase is subject to severe substrate inhibition. PMID- 10838563 TI - How do presynaptic PLA2 neurotoxins block nerve terminals? AB - Snake presynaptic neurotoxins with phospholipase A2 activity block nerve terminals in an unknown way. Here, we propose that they enter the lumen of synaptic vesicles following endocytosis and hydrolyse phospholipids of the inner leaflet of the membrane. The transmembrane pH gradient drives the translocation of fatty acids to the cytosolic monolayer, leaving lysophospholipids on the lumenal layer. Such vesicles are highly fusogenic and release neurotransmitter upon fusion with the presynaptic membrane, but cannot be retrieved because of the high local concentration of fatty acids and lysophospholipids, which prevents vesicle neck closure. PMID- 10838564 TI - Asparagine repeats are rare in mammalian proteins. PMID- 10838565 TI - Functionally unrelated signalling proteins contain a fold similar to Mg2+ dependent endonucleases. PMID- 10838566 TI - Evidence of domain swapping within the jumonji family of transcription factors. PMID- 10838567 TI - Transcriptional regulation through Mediator-like coactivators in yeast and metazoan cells. AB - A novel multiprotein complex has recently been identified as a coactivator for transcriptional control of protein-encoding genes by RNA polymerase II in higher eukaryotic cells. This complex is evolutionarily related to the Mediator complex from yeast and, on the basis of its structural and functional characteristics, promises to be a key target of diverse regulatory circuits. PMID- 10838568 TI - LNB-TM7, a group of seven-transmembrane proteins related to family-B G-protein coupled receptors. AB - A number of unusual seven-transmembrane molecules have recently been characterized that have significant amino acid sequence similarity within the membrane-spanning hydrophobic regions and intervening loops to members of G protein-coupled receptor family B. However, in contrast to the family-B G-protein coupled receptors, these molecules have unusually large N-terminal extracellular domains that contain a number of well- characterized protein modules. The range of cell types expressing these complex molecules and their potential roles in cell adhesion and signalling have become a major focus of research in a number of biological systems. PMID- 10838569 TI - Connecting transcription to messenger RNA processing. AB - The production of messenger RNA by gene transcription requires at least three RNA processing mechanisms: capping, splicing and polyadenylation. All three reactions occur in intimate association with the elongating polymerase complex through the C terminus of the largest subunit of RNA polymerase II. The processing of mRNA is therefore orchestrated to act on the nascent RNA as soon as it emerges from the polymerase complex. PMID- 10838570 TI - ARID proteins come in from the desert. AB - Members of the recently discovered ARID (AT-rich interaction domain) family of DNA-binding proteins are found in fungi and invertebrate and vertebrate metazoans. ARID-encoding genes are involved in a variety of biological processes including embryonic development, cell lineage gene regulation and cell cycle control. Although the specific roles of this domain and of ARID-containing proteins in transcriptional regulation are yet to be elucidated, they include both positive and negative transcriptional regulation and a likely involvement in the modification of chromatin structure. PMID- 10838571 TI - The bioinformatics resource PMID- 10838572 TI - Cn3D: sequence and structure views for Entrez. PMID- 10838573 TI - From 'reactive C2 units' to acetyl coenzyme A: a long trail with an acetyl phosphate detour. PMID- 10838574 TI - Peptides, Pseudomonas aeruginosa, polysaccharides and lipopolysaccharides- players in the predicament of cystic fibrosis patients. PMID- 10838575 TI - Peptides, pseudomonas aeruginosa, polysaccharides and lipopolysaccharides - players in the predicament of cystic fibrosis patients: response PMID- 10838577 TI - The dynamics of receptor recognition by human rhinoviruses: response PMID- 10838576 TI - The dynamics of receptor recognition by human rhinoviruses. PMID- 10838578 TI - The role of Chlamydia pneumoniae in atherosclerosis--recent evidence from animal models. PMID- 10838579 TI - The role of chlamydia pneumoniae in atherosclerosis - recent evidence from animal models: response PMID- 10838581 TI - The structural basis of CEACAM-receptor targeting by neisserial opa proteins: response PMID- 10838580 TI - The structural basis of CEACAM-receptor targeting by neisserial Opa proteins. PMID- 10838582 TI - Infectious clones of herpesviruses: a new approach for understanding viral gene function. PMID- 10838583 TI - Measuring virulence without a target. PMID- 10838584 TI - Playing second fiddle: second-strand processing and liberation of transposable elements from donor DNA. AB - Retroviruses and many transposons of both prokaryotes and eukaryotes share similar chemical reactions in their transposition. Some elements remain attached to donor DNA during transposition and their translocation results in a fusion between target and donor replicons. However, many elements are separated from their flanking donor DNA prior to their insertion into a target site, which requires processing of both strands at both ends of the element. A variety of strategies have been adopted for cleavage of the second, complementary strand to liberate the transposon. PMID- 10838585 TI - The bactericidal action of penicillin: new clues to an unsolved mystery. AB - The Staphylococcus aureus lrgAB operon was recently shown to inhibit extracellular murein hydrolase activity and increase tolerance to penicillin. Further characterization of this operon could provide novel insight into the dynamics of S. aureus cell wall metabolism and the mechanism of penicillin induced lethality. PMID- 10838586 TI - Chromosome replication, nucleoid segregation and cell division in archaea. AB - Recent progress in cell cycle analysis of archaea has included the identification of putative chromosome replication origins, novel DNA polymerases and an unusual mode of cell cycle organization featuring multiple copies of the chromosome and asymmetric cell divisions. Genome sequence data indicate that in crenarchaea, the 'ubiquitous' FtsZ/MinD-based prokaryotic cell division apparatus is absent and division therefore must occur by unique, as-yet-unidentified mechanisms. The evolutionary and functional relationships between the archaeal Cdc6 protein and bacterial and eukaryal replication initiation factors are discussed. PMID- 10838587 TI - Why aren't foamy viruses pathogenic? AB - Foamy viruses are complex retroviruses that lead to either highly cytopathic or persistent infections in vitro, but to non-pathogenic lifelong infections in naturally or accidentally infected hosts. Factors that could contribute to these benign persistent infections include regulated transcription from the two viral promoters, the functions of the Bet accessory protein and the host immune response. PMID- 10838588 TI - Potassium-channel closure taken to TASK. PMID- 10838589 TI - Protein targeting: altering receptor kinase function in the brain. AB - Gene targeting has proved to be one of the most powerful techniques with which one can investigate molecular mechanisms that underlie complex phenomena such as learning and memory. Despite its popularity, however, concerns have been raised about this technique and alternative approaches have been sought. One such approach is protein targeting, which is based on the application of immunoadhesins, genetically engineered fusion proteins that exhibit functionally relevant target specificity. These immunoadhesins modulate the activity of not only a single receptor but of all receptors with homologous binding sites, which thereby eliminates the possibility of compensation by sister receptors. Furthermore, immunoadhesins can be used not only to impair but also to improve receptor function in the brain. Initial studies using immunoadhesins suggest that protein targeting might be a useful approach for analyzing the molecular mechanisms of brain function and behavior. PMID- 10838590 TI - Neuronal cell death in Huntington's disease: a potential role for dopamine. AB - Huntington's disease is an inherited neurodegenerative disorder, the cause of which is unknown. Excitotoxicity, mitochondrial dysfunction and oxidative stress are all likely to contribute to the striatal cell death that occurs in this disorder. There are accumulating data indicating that under specific circumstances, dopamine, which occurs in high concentrations in the basal ganglia, might be neurotoxic. In this article, the current models used to study Huntington's disease are reviewed and the recent findings that implicate dopamine in the pathophysiology of this progressive disorder are discussed. Although many questions remain unanswered, the dopaminergic system could contribute to striatal vulnerability in Huntington's disease and provide a novel avenue for the development of new therapies. PMID- 10838591 TI - Clinical trials with neuroprotective drugs in acute ischaemic stroke: are we doing the right thing? PMID- 10838592 TI - Reply PMID- 10838593 TI - Low-level memory processes in vision. AB - Psychophysical studies of the short-term memory for attributes or dimensions of the visual stimulus that are known to be important in early visual processing (spatial frequency, orientation, contrast, motion and color) identify a low-level perceptual memory mechanism. This proposed mechanism is located early in the visual processing stream, prior to the structural description system responsible for shape priming but beyond primary visual cortex (V1); it is composed of a series of parallel, special-purpose perceptual mechanisms with independent but limited processing resources. Each mechanism is devoted to the analysis of a single dimension and is coupled to a memory store. PMID- 10838594 TI - Seasonal plasticity in the adult brain. AB - Seasonal plasticity of structure and function is a fundamental feature of nervous systems in a wide variety of animals that occupy seasonal environments. Excellent examples of seasonal brain changes are found in the avian song control system, which has become a leading model of morphological and functional plasticity in the adult CNS. The volumes of entire brain regions that control song increase dramatically in anticipation of the breeding season. These volumetric changes are induced primarily by vernal increases in circulating sex steroids and are accompanied by increases in neuronal size, number and spacing. In several species, these structural changes in the song control circuitry are associated with seasonal changes in song production and learning. Songbirds provide important insights into the mechanisms and behavioral consequences of plasticity in the adult brain. PMID- 10838595 TI - Cytosolic phospholipase A2 and the distinct transcriptional programs of astrocytoma cells. AB - Astrocytes constitute the most abundant cell type in the nervous system. Under physiological conditions, they respond to the stimuli to which neurons are also responsive. The use of astrocytoma cell lines with well-defined morphological and functional markers has been helpful for addressing the mechanisms of signal transduction that operate in the nervous system. On the basis of the effects produced by agonists of different types of receptor (muscarinic ACh receptors, thrombin receptors, phospholipases A2 receptors and tumor necrosis factor alpha receptors), several different transcriptional programs that involve the MAP kinase-cytosolic phospholipase A2 system and the transcription factor NF-kappaB have been described. PMID- 10838596 TI - Recovery recapitulates ontogeny. AB - Several studies support the hypothesis that after stroke, specific features of brain function revert to those seen at an early stage of development, with the subsequent process of recovery recapitulating ontogeny in many ways. Many clinical characteristics of stroke recovery resemble normal development, particularly in the motor system. Consistent with this, brain-mapping studies after an ischemic insult suggest re-emergence of childhood organizational patterns: recovery being associated with a return to adult patterns. Experimental animal studies demonstrate increased levels of developmental proteins, particularly in the area surrounding an infarct, suggesting an active process of reconditioning in response to cerebral ischemia. Understanding the patterns of similarity between normal development and stroke recovery might be of value in its treatment. PMID- 10838597 TI - Arbitrary associations between antecedents and actions. AB - The arbitrary linkage of sensory cues to actions and goals represents one of the most-flexible capabilities in the behavioral repertoire of mammals. This ability has been termed 'conditional motor learning', 'conditional discrimination' or, more recently, 'arbitrary visuomotor mapping'. Unlike other forms of visuomotor guidance, in arbitrary mapping the location of the sensory cue lacks any systematic spatial relationship with the action or its goal. Recent work has identified much of the neural network that underlies this behavior. It consists of parts of the frontal cortex, hippocampal system and basal ganglia, each of which has neurons whose activity undergoes systematic evolution during learning. PMID- 10838598 TI - Endocannabinoids and multiple sclerosis: a blessing from the 'inner bliss'? PMID- 10838599 TI - Reply: a sanguine approach to cannabis PMID- 10838600 TI - Schizophrenia, the dopamine hypothesis and alpha2-adrenoceptor antagonists. PMID- 10838601 TI - Maximal function of minimal K+ channel subunits. PMID- 10838602 TI - Central 5-HT1A receptors and vagal tone to the airways. PMID- 10838603 TI - Reply: 5-HT1A receptors and control of airways PMID- 10838604 TI - Anandamide and the question of its functional role for activation of capsaicin receptors. PMID- 10838605 TI - Zinc and immunoresistance to infection in aging: new biological tools. AB - Infections can cause mortality when the immune system is damaged. The catalytic, structural (in zinc-finger proteins) and regulatory roles of zinc mean that this ion is involved in the maintenance of an effective immune response. Both zinc deficiency and impaired cell-mediated immunity combine during aging to result in increased susceptibility to infection. Dietary supplementation with the recommended daily allowance of zinc for between one and two months decreases the incidence of infection and increases the survival rate following infection in the elderly. This article reviews the biochemical pathways through which zinc might act to increase immunoresistance to infection in the elderly. PMID- 10838606 TI - The contribution of pharmacology to research on the mechanisms of memory formation. AB - Pharmacological studies of memory are motivated by the hope or hypothesis that behavioural findings, considered together with knowledge of the mechanisms of drug action, will help to elucidate the neurobiological bases of memory. There is now considerable evidence that this hope is justified. PMID- 10838608 TI - Nicotinic receptor function: new perspectives from knockout mice. AB - Knockout mice, in which one or more genes of interest are silenced, provide unique opportunities to analyse diverse aspects of gene function in vivo. In particular, the contribution of the encoded protein(s) in complex behaviours can be assessed. Since the first targeted disruption in 1995 of the gene encoding the beta2-subunit of the nicotinic acetylcholine receptor (nAChR), all but a few of the mammalian nAChR subunits have been disrupted (i.e. alpha7, alpha4, alpha3, alpha9, beta4 and beta3). Recent advances brought by genetically modified mice to our understanding of the endogenous composition and role of nAChRs in the nervous system, and of the diverse pharmacological actions of nicotine regarding learning, analgesia, reinforcement, development and aging in the brain will be discussed. PMID- 10838609 TI - The endocannabinoid system as a target for therapeutic drugs. AB - Cannabinoid receptors, the molecular targets of the cannabis constituent Delta9 tetrahydrocannabinol, are present throughout the body and are normally bound by a family of endogenous lipids - the endocannabinoids. Release of endocannabinoids is stimulated in a receptor-dependent manner by neurotransmitters and requires the enzymatic cleavage of phospholipid precursors present in the membranes of neurons and other cells. Once released, the endocannabinoids activate cannabinoid receptors on nearby cells and are rapidly inactivated by transport and subsequent enzymatic hydrolysis. These compounds might act near their site of synthesis to serve a variety of regulatory functions, some of which are now beginning to be understood. Recent advances in the biochemistry and pharmacology of the endocannabinoid system in relation to the opportunities that this system offers for the development of novel therapeutic agents will be discussed. PMID- 10838611 TI - The response of epidermal cells to contact. PMID- 10838610 TI - Ischaemic preconditioning of the vasculature: an overlooked phenomenon for protecting the heart? AB - Exposing the heart to brief episodes of ischaemia protects the myocardium and vascular endothelial cells against functional damage and cell death caused by subsequent prolonged ischaemia. Elucidation of the mechanisms that are involved in this phenomenon known as 'ischaemic preconditioning' and identification of drugs that mimic the protective response have the potential to improve the prognosis of myocardial infarction and other cardiac syndromes dramatically. This article focuses on recent findings on the effects of ischaemic preconditioning of the coronary vasculature, which highlight the endothelium as an important target for a successful therapeutic approach to myocardial ischaemia-reperfusion injury. PMID- 10838612 TI - Breeding mercury-breathing plants for environmental cleanup. AB - In an elegant study, Richard Meagher's research group has succeeded in introducing a bacterial mercury detoxification pathway into plants. The resulting plants show enhanced mercury tolerance and can convert highly toxic forms of organic mercury to less toxic elemental mercury, which is volatile. The significance of this study is that it could lead to the more efficient and affordable cleanup of environmental mercury pollution, and in a broader context, it proves the power of genetic engineering for phytoremediation. PMID- 10838613 TI - The retinoblastoma protein - a bridge to heterochromatin. PMID- 10838614 TI - The role of calcium and activated oxygens as signals for controlling cross tolerance. AB - Plants are confronted on a regular basis with a range of environmental stresses. These include abiotic insults caused by, for example, extreme temperatures, altered water status or nutrients, and biotic stresses generated by a plethora of plant pathogens. Many studies have shown that the cellular responses to these environmental challenges are rather similar, which might be why plants resistant to one stress are sometimes cross-tolerant to others. To understand this phenomenon and to be able to take full advantage of it in agriculture, we must determine whether the individual biochemical pathways that make up the responses to each external stimulus are activated by unique, overlapping or redundant signalling systems. We discuss the potential role of signalling molecules, such as calcium and activated oxygen species, in underlying cross-tolerance. PMID- 10838615 TI - Colinearity and gene density in grass genomes. AB - Grasses are the single most important plant family in agriculture. In the past years, comparative genetic mapping has revealed conserved gene order (colinearity) among many grass species. Recently, the first studies at gene level have demonstrated that microcolinearity of genes is less conserved: small scale rearrangements and deletions complicate the microcolinearity between closely related species, such as sorghum and maize, but also between rice and other crop plants. In spite of these problems, rice remains the model plant for grasses as there is limited useful colinearity between Arabidopsis and grasses. However, studies in rice have to be complemented by more intensive genetic work on grass species with large genomes (maize, Triticeae). Gene-rich chromosomal regions in species with large genomes, such as wheat, have a high gene density and are ideal targets for partial genome sequencing. PMID- 10838616 TI - Inositol signaling and plant growth. AB - Living organisms have evolved to contain a wide variety of receptors and signaling pathways that are essential for their survival in a changing environment. Of these, the phosphoinositide pathway is one of the best conserved. The ability of the phosphoinositides to permeate both hydrophobic and hydrophilic environments, and their diverse functions within cells have contributed to their persistence in nature. In eukaryotes, phosphoinositides are essential metabolites as well as labile messengers that regulate cellular physiology while traveling within and between cells. The stereospecificity of the six hydroxyls on the inositol ring provides the basis for the functional diversity of the phosphorylated isomers that, in turn, generate a selective means of intracellular and intercellular communication for coordinating cell growth. Although such complexity presents a difficult challenge for bench scientists, it is ideal for the regulation of cellular functions in living organisms. PMID- 10838617 TI - Patchy stomatal conductance: emergent collective behaviour of stomata. AB - Until recently, most scientists have tacitly assumed that individual stomata respond independently and similarly to stimuli, showing minor random variation in aperture and behaviour. This implies that stomatal behaviour should not depend on the scale of observation. However, it is now clear that these assumptions are often incorrect. Leaves frequently exhibit dramatic spatial and temporal heterogeneity in stomatal behaviour. This phenomenon, in which small 'patches' of stomata respond differently from those in adjacent regions of the leaf, is called 'patchy stomatal conductance'. It appears to represent a hitherto unknown type of emergent collective behaviour that manifests itself in populations of stomata in intact leaves. PMID- 10838618 TI - Transport and action of ascorbate at the plant plasma membrane. AB - The plasmalemma is both a bridge and a barrier between the cytoplasm and the outside world. It is a dynamic interface that perceives and transmits information concerning changes in the environment to the nucleus to modify gene expression. In plants, ascorbate is an essential part of this dialogue. The concentration and ratio of reduced to oxidized ascorbate in the apoplast, for example, possibly modulates cell division and growth. The leaf apoplast contains millimolar amounts of ascorbate that protect the plasmalemma against oxidative damage. The apoplastic ascorbate-dehydroascorbate redox couple is linked to the cytoplasmic ascorbate-dehydroascorbate redox couple by specific transporters for either or both metabolites. Although evidence about the mechanisms driving ascorbate or dehydroascorbate transport remains inconclusive, these carrier proteins potentially regulate the level and redox status of ascorbate in the apoplast. The redox coupling between compartments facilitated by these transport systems allows coordinated control of key physiological responses to environmental cues. PMID- 10838619 TI - A piece of my mind: paying attention. PMID- 10838620 TI - JAMA 100 years ago: treatment of measles with red light PMID- 10838622 TI - JAMA 100 years ago: treatment of scarlatinal nephritis PMID- 10838621 TI - JAMA 100 years ago: bathing treatment in chronic kidney affections PMID- 10838624 TI - JAMA 100 years ago: treatment of valve pneumothorax PMID- 10838623 TI - JAMA 100 years ago: methylene blue in grave malaria cachexia PMID- 10838625 TI - Will evidence-based practice help span gulf between medicine and law? PMID- 10838626 TI - IOM report a blueprint for elimination of TB. Institute of Medicine. PMID- 10838627 TI - Once again, 140/90 is the goal. PMID- 10838628 TI - Medicine in uniform: turnover at the Top PMID- 10838629 TI - Medicine in uniform: improved medical helicopter PMID- 10838630 TI - Medicine in uniform: getting enough ZZZs PMID- 10838631 TI - Medicine in uniform: teams terror-ready PMID- 10838632 TI - Medicine in uniform: battle of mogadishu PMID- 10838633 TI - Improving the conduct and reporting of clinical trials. PMID- 10838634 TI - Improving the conduct and reporting of clinical trials. PMID- 10838635 TI - Improving the conduct and reporting of clinical trials. PMID- 10838636 TI - Improving the conduct and reporting of clinical trials. PMID- 10838637 TI - Improving the conduct and reporting of clinical trials PMID- 10838638 TI - Improving the conduct and reporting of clinical trials PMID- 10838639 TI - Out-of-hospital endotracheal intubation of children. PMID- 10838640 TI - Out-of-hospital endotracheal intubation of children. PMID- 10838641 TI - Out-of-hospital endotracheal intubation of children. PMID- 10838643 TI - Out-of-hospital endotracheal intubation of children PMID- 10838642 TI - Out-of-hospital endotracheal intubation of children. PMID- 10838644 TI - Evidence-based assessment of medical technology. PMID- 10838645 TI - Evidence-based assessment of medical technology PMID- 10838646 TI - False-positive tuberculin skin test results among health care workers. PMID- 10838647 TI - Safety, immunogenicity, and induction of immunologic memory by a serogroup C meningococcal conjugate vaccine in infants: A randomized controlled trial. AB - CONTEXT: Neisseria meningitidis is a common cause of meningitis and septicemia in infants worldwide. Whether a meningococcal C conjugate vaccine protects infants against the serogroup C strain is unknown. OBJECTIVES: To determine whether a meningococcal C conjugate vaccine is safe and immunogenic and induces immunologic memory in infants. DESIGN: Single-center, double-blind, randomized controlled trial in 1995 and 1996. SETTING: Community, Oxfordshire, England. PARTICIPANTS: One hundred eighty-two healthy infants. INTERVENTIONS: Participants were randomly assigned to receive vaccination with 0. 5-mL doses of 1 of 2 lots of meningococcal C conjugate vaccine (groups 1 and 2; n=60 in each group) or a hepatitis B control vaccine (group 3; n=62), administered with routine immunizations at 2, 3, and 4 months of age. Approximately half of each group received meningococcal C conjugate vaccine and half received plain meningococcal polysaccharide vaccine (MPS) at 12 months of age. MAIN OUTCOME MEASURES: Serum antibodies to meningococcal C polysaccharide, assayed by enzyme-linked immunosorbent assay, and serum bactericidal activity (SBA), at 2, 3, 4, 5, 12, and 13 months of age; local and systemic reactions, recorded for 6 days after each vaccination, compared by intervention group. RESULTS: Meningococcal C conjugate vaccine was well tolerated. After 3 doses, children in groups 1 and 2 achieved significantly higher meningococcal C IgG geometric mean concentrations (21 and 17 U/mL, respectively, vs 0.20 U/mL; P<.001) and SBA titers (629 and 420, respectively, vs 4.1; P<. 001) than controls. At 12 months, antibody concentrations had decreased in all groups but remained significantly higher in children vaccinated with meningococcal C conjugate vaccine (SBA, 24 and 16 in groups 1 and 2, respectively, vs 4.2 in group 3; P<.001). Following vaccination with MPS at 12 months of age, SBA in the meningococcal C conjugate vaccine group was significantly higher than in controls (SBA, 789 vs 4.5; P<.001). CONCLUSIONS: Our data indicate that meningococcal C conjugate vaccine is safe and immunogenic and results in immunologic memory when given with other routinely administered vaccines to infants at 2, 3, and 4 months of age. JAMA. 2000;283:2795-2801 PMID- 10838648 TI - Effectiveness of a nurse-based outreach program for identifying and treating psychiatric illness in the elderly. AB - CONTEXT: Elderly persons with psychiatric disorders are less likely than younger adults to be diagnosed as having a mental disorder and receive needed mental health treatment. Lack of access to care is 1 possible cause of this disparity. OBJECTIVE: To determine whether a nurse-based mobile outreach program to seriously mentally ill elderly persons is more effective than usual care in diminishing levels of depression, psychiatric symptoms, and undesirable moves (eg, nursing home placement, eviction, board and care placement). DESIGN: Prospective randomized trial conducted between March 1993 and April 1996 to assess the effectiveness of the Psychogeriatric Assessment and Treatment in City Housing (PATCH) program. SETTING: Six urban public housing sites for elderly persons in Baltimore, Md. PARTICIPANTS: A total of 945 (83%) of 1195 residents in the 6 sites underwent screening for psychiatric illness. Among those screened, 342 screened positive and 603 screened negative. All screen-positive subjects aged 60 years and older (n=310) and a 10% random sample of screen-negative subjects aged 60 years and older (n=61) were selected for a structured psychiatric interview. Eleven subjects moved or died; 245 (82%) of those who screened positive and 53 (88%) of those who screened negative were evaluated to determine who had a psychiatric disorder. Data were weighted to estimate the prevalence of psychiatric disorders at the 6 sites. INTERVENTION: Among the 6 sites, residents in 3 buildings were randomized to receive the PATCH model intervention, which included educating building staff to be case finders, performing assessment in residents' apartments, and providing care when indicated; and residents in the remaining 3 buildings were randomized to receive usual care (comparison group). MAIN OUTCOME MEASURES: Number of undesirable moves and scores on the Montgomery-Asberg Depression Rating Scale (MADRS), a measure of depressive symptoms, and the Brief Psychiatric Rating Scale (BPRS), a measure of psychiatric symptoms and behavioral disorder, in intervention vs comparison sites. RESULTS: Based on weighted data, at 26 months of follow-up, psychiatric cases at the intervention sites had significantly lower (F(1)=31.18; P<.001) MADRS scores (9.1 vs 15.2) and significantly lower (F(1)=17.35; P<.001) BPRS scores (27.4 vs 33.9) than those at the nontreatment comparison sites. There was no significant difference between the groups in undesirable moves (relative risk, 0.97; 95% confidence interval, 0. 44-2.17). CONCLUSIONS: These results indicate that the PATCH intervention was more effective than usual care in reducing psychiatric symptoms in persons with psychiatric disorders and those with elevated levels of psychiatric symptoms. JAMA. 2000;283:2802-2809 PMID- 10838649 TI - Calcification of the aortic arch: risk factors and association with coronary heart disease, stroke, and peripheral vascular disease. AB - CONTEXT: Calcium deposits in coronary and extracoronary arterial beds may indicate the extent of atherosclerosis. However, the incremental predictive value of vascular calcification, beyond traditional coronary risk factors, is not clearly established. OBJECTIVE: To evaluate risk factors for aortic arch calcification and its long-term association with cardiovascular diseases in a population-based sample. DESIGN AND SETTING: Cohort study conducted at a health maintenance organization in northern California. PARTICIPANTS: A total of 60,393 women and 55,916 men, aged 30 to 89 years at baseline who attended multiphasic health checkups between 1964 and 1973 and for whom incidence of hospitalizations and/or mortality data were ascertained using discharge diagnosis codes and death records through December 31, 1997 (median follow-up, 28 years). MAIN OUTCOME MEASURE: Hospitalization for or death due to coronary heart disease, ischemic stroke, hemorrhagic stroke, or peripheral vascular disease, as associated with aortic arch calcification found on chest radiograph at checkup from 1964-1973. RESULTS: Aortic arch calcification was present in 1.9% of men and 2.6% of women. It was independently associated with older age, no college education, current smoking, and hypertension in both sexes, but it was inversely related to body mass index and family history of myocardial infarction. In women, aortic arch calcification was also associated with black race and elevated serum cholesterol level. After adjustment for age, educational attainment, race/ethnicity, cigarette smoking, alcohol consumption, body mass index, serum cholesterol level, hypertension, diabetes, and family history of myocardial infarction, aortic arch calcification was associated with an increased risk of coronary heart disease (in men, relative risk [RR], 1.27; 95% confidence interval [CI], 1.11-1.45; in women, RR, 1. 22; 95% CI, 1.07-1.38). Among women, it was also independently associated with a 1.46-fold increased risk of ischemic stroke (95% CI, 1.28-1.67). CONCLUSION: In our population-based cohort, aortic arch calcification was independently related to coronary heart disease risk in both sexes as well as to ischemic stroke risk in women. JAMA. 2000;283:2810-2815 PMID- 10838650 TI - A clinical decision support system for prevention of venous thromboembolism: effect on physician behavior. AB - CONTEXT: Computer-based clinical decision support systems (CDSSs) have been promoted for their potential to improve quality of health care. However, given the limited range of clinical settings in which they have been tested, such systems must be evaluated rigorously before widespread introduction into clinical practice. OBJECTIVE: To determine whether presentation of venous thromboembolism prophylaxis guidelines using a CDSS increases the proportion of appropriate clinical practice decisions made. DESIGN: Time-series study conducted between December 1997 and July 1999. SETTING: Orthopedic surgery department of a teaching hospital in Paris, France. PARTICIPANTS: A total of 1971 patients who underwent orthopedic surgery. INTERVENTION: A CDSS designed to provide immediate information pertaining to venous thromboembolism prevention among surgical patients was integrated into daily medical practice during three 10-week intervention periods, alternated with four 10-week control periods, with a 4-week washout between each period. MAIN OUTCOME MEASURE: Proportion of appropriate prescriptions ordered for anticoagulation, according to preestablished clinical guidelines, during intervention vs control periods. RESULTS: Physicians complied with guidelines in 82.8% (95% confidence interval [CI], 77.6%-87.1%) of cases during control periods and in 94.9% (95% CI, 92.5%-96.6%) of cases during intervention periods. During each intervention period, the appropriateness of prescription increased significantly (P<.001). Each time the CDSS was removed, physician practice reverted to that observed before initiation of the intervention. The relative risk of inappropriate practice decisions during control periods vs intervention periods was 3.8 (95% CI, 2.7-5.4). CONCLUSIONS: In our study, implementation of clinical guidelines for venous thromboembolism prophylaxis through a CDSS used routinely in an orthopedic surgery department and integrated into the hospital information system changed physician behavior and improved compliance with guidelines. JAMA. 2000;283:2816-2821 PMID- 10838651 TI - Effect of calcium carbonate on the absorption of levothyroxine. AB - CONTEXT: The effect of calcium carbonate on the absorption of levothyroxine has not been studied systematically. Such a potential drug interaction merits investigation because concurrent treatment with both drugs is common, particularly in postmenopausal women. OBJECTIVE: To investigate the potential interference of calcium carbonate in the absorption of levothyroxine. DESIGN: Prospective cohort study conducted from November 1998 to June 1999, supplemented with an in vitro study of thyroxine (T(4)) binding to calcium carbonate. SETTING: Veterans Affairs Medical Center in West Los Angeles, Calif. PATIENTS: Twenty patients (age range, 27-78 years; n=11 men) with hypothyroidism who were taking a stable long-term regimen of levothyroxine were included in the study. All patients had serum free T(4) and thyrotropin values in the normal range before beginning the study. INTERVENTION: Subjects were instructed to take 1200 mg/d of elemental calcium as calcium carbonate, ingested with their levothyroxine, for 3 months. MAIN OUTCOME MEASURES: Levels of free T(4), total T(4), total triiodothyronine (T(3)), and thyrotropin, measured in all subjects at baseline (while taking levothyroxine alone), at 2 and 3 months (while taking calcium carbonate and levothyroxine), and 2 months after calcium carbonate discontinuation (while continuing to take levothyroxine). RESULTS: Mean free T(4) and total T(4) levels were significantly reduced during the calcium period and increased after calcium discontinuation. Mean free T(4) levels were 17 pmol/L (1.3 ng/dL) at baseline, 15 pmol/L (1.2 ng/dL) during the calcium period, and 18 pmol/L (1.4 ng/dL) after calcium discontinuation (overall P<.001); mean total T(4) levels were 118 nmol/L (9.2 microg/dL) at baseline, 111 nmol/L (8.6 microg/dL) during the calcium period, and 120 nmol/L (9.3 microg/dL) after calcium discontinuation (overall P=.03). Mean thyrotropin levels increased significantly, from 1.6 mIU/L at baseline to 2.7 mIU/L during the calcium period, and decreased to 1. 4 mIU/L after calcium discontinuation (P=.008). Twenty percent of patients had serum thyrotropin levels higher than the normal range during the calcium period; the highest observed level was 7.8 mIU/L. Mean T(3) levels did not change during the calcium period. The in vitro study of T(4) binding to calcium showed that adsorption of T(4) to calcium carbonate occurs at acidic pH levels. CONCLUSIONS: This study of 20 patients receiving long-term levothyroxine replacement therapy indicates that calcium carbonate reduces T(4) absorption and increases serum thyrotropin levels. Levothyroxine adsorbs to calcium carbonate in an acidic environment, which may reduce its bioavailability. JAMA. 2000;283:2822-2825 PMID- 10838652 TI - Effectiveness of automatic shoulder belt systems in motor vehicle crashes. AB - CONTEXT: Approximately 10 million cars with automatic shoulder belt systems are currently in use in the United States. However, reports on the effectiveness of such restraints have yielded conflicting results. OBJECTIVE: To determine the effectiveness of automatic shoulder belt systems in reducing the risk of injury and death among front-seat passenger vehicle occupants. DESIGN, SETTING, AND SUBJECTS: Analysis of data collected from the 1993-1996 National Highway Traffic Safety Administration Crashworthiness Data System on front-seat occupants involved in 25,811 tow-away crashes of passenger cars, light trucks, vans, and sport utility vehicles. MAIN OUTCOME MEASURES: Death and serious injury to specific body areas by use of manual lap and shoulder belts, automatic shoulder belts with manual lap belts, or automatic shoulder belts without lap belts, compared with no restraint use. RESULTS: Use of automatic shoulder belts without lap belts was associated with a decrease in the risk of death vs no restraint use but was not statistically significant for all crashes (odds ratio [OR], 0.66; 95% confidence interval [CI], 0.42-1.06) or for frontal crashes (OR, 0.71; 95% CI, 0.38-1.35) after adjustment for occupant age, sex, vehicle year, air-bag deployment, estimated change in vehicle speed during the crash, and principal direction of force. This association was significantly weaker than the 86% lower risk observed for use of automatic shoulder belts with lap belts (OR, 0.14; 95% CI, 0.07-0.26 vs no restraint; P<.05). Use of automatic shoulder belts without lap belts was associated with an increased risk of serious chest (OR, 2.66; 95% CI, 1.11-6.35) and abdominal (OR, 2.06; 95% CI, 1.004-4.22) injuries for all crashes. CONCLUSIONS: These data indicate that improperly used automatic restraint systems may be less effective than properly used systems and are associated with an increased risk of serious chest and abdominal injuries. Given the continued widespread use of these automatic systems, educational programs may be warranted. JAMA. 2000;283:2826-2828 PMID- 10838653 TI - Users' guides to the medical literature: XX. Integrating research evidence with the care of the individual patient. Evidence-Based Medicine Working Group. AB - Clinicians can use research results to determine optimal care for an individual patient by using a patient's baseline risk estimate, clinical prediction guidelines that quantitate an individual patient's potential for benefit, and published articles. We propose that when clinicians are determining the likelihood that treatment will prevent the target event (at the expense of adverse events) in a patient that they also incorporate the patient's values. The 3 main elements to joint clinical decision making are disclosure of information about the risks and benefits of therapeutic alternatives, exploration of the patient's values about both the therapy and potential outcomes, and the actual decision. In addressing the patient's risk of adverse events without treatment and risk of harm with therapy, clinicians must recognize that patients are rarely identical to the average study patient. Differences between study participants and patients in real-world practice tend to be quantitative (differences in degree of risk of the outcome or responsiveness to therapy) rather than qualitative (no risk or adverse response to therapy). The number needed to treat and number needed to harm can be used to generate patient-specific estimates relative to the risk of the outcome event. Clinicians must consider a patient's risk of adverse events from any intervention and incorporate the patient's values in clinical decision making by using information about the risks and benefits of therapeutic alternatives. JAMA. 2000;283:2829-2836 PMID- 10838654 TI - Public health law in a new century: part I: law as a tool to advance the community's health. AB - Statutes, regulations, and litigation are pivotal tools for creating conditions for people to lead healthier and safer lives. Law can educate, create incentives, and deter; mandate safer product design and use of property; and alter the informational, physical, or economic environment. This article defines public health law as the power and duty of the state to ensure conditions for people to be healthy and limitations on the state's power to constrain autonomy, privacy, liberty, and proprietary interests of individuals and businesses. The 5 essential characteristics of public health law discussed are (1) the government's responsibility to defend against health risks and promote the public's health; (2) the population-based perspective of public health, emphasizing prevention; (3) the relationship between government and the populace; (4) the mission, core functions, and services of the public health system; and (5) the power to coerce individuals, professionals, and businesses for the community's protection. JAMA. 2000;283:2837-2841 PMID- 10838655 TI - New opportunities for prevention of meningococcal disease. PMID- 10838656 TI - The public health model for mental health care for the elderly. PMID- 10838657 TI - Quantification of tumor-specific T lymphocytes with the ELISPOT assay. AB - The characterization of tumor-associated antigens and of human leukocyte antigen (HLA) class I molecule-binding peptide epitopes derived from these antigens has prompted the initiation of various vaccination trials aimed at inducing tumor specific CD8+ T cells in persons with cancer. Sensitive and easy-to-perform T cell assays that assess the frequency of tumor-reactive T cells are crucial for the evaluation and further development of vaccination approaches. This review focuses on a novel ELISPOT technique that allows quantification of tumor-specific T lymphocytes from peripheral blood by detecting antigen-induced cytokine secretion. Various ELISPOT methods using different antigen-presenting cells and different cytokines as read-out are described. T-cell analyses performed using the standard chromium release assay and the ELISPOT assay are also compared. Results from various clinical trials, including peptide and whole tumor cell vaccination and cytokine treatment, are now available and show the suitability of the ELISPOT assay for monitoring T-cell responses. To establish a basis for standardization and to further improve this technique, the first comparative quality assurance studies analyzing T-cell frequencies in different laboratories with the ELISPOT assay are being performed. PMID- 10838658 TI - Induction of human leukocyte antigen-A26-restricted and tumor-specific cytotoxic T lymphocytes by a single peptide of the SART1 antigen in patients with cancer with different A26 subtypes. AB - Peptide antigens available for use in specific immunotherapy of patients with cancer have not been fully determined. Although the authors have reported the SART1 gene encoding epitopes recognized by HLA-A2601-restricted and tumor specific cytotoxic T lymphocytes (CTLs), the HLA-A26 allele is mainly subdivided into A2601, A2602, and A2603 subtypes. In this study, the authors attempted to determine whether the SART1-derived peptide at position 736-744 (KGSGKMKTE) is suitable to induce HLA-A26-restricted and tumor-specific CTLs in patients with cancer who have these subtypes. This peptide induced the HLA-A26 subtype restricted and tumor-specific CTLs in HLA-A2601+ or HLA-A2603+ peripheral blood mononuclear cells, respectively. It also induced the HLA-A26-restricted CTL activity in HLA-A2602+ peripheral blood mononuclear cells. Therefore, this peptide could be useful for specific immunotherapy of patients with cancer who have any of the three HLA-A26 subtypes. PMID- 10838659 TI - Targeting of natural killer-like T immunologic effector cells against leukemia and lymphoma cells by reverse antibody-dependent cellular cytotoxicity. AB - Recently, highly efficient natural killer-like T immunologic effector cells called cytokine-induced killer (CIK) cells have been described. Most interestingly, CIK cells have been shown to eradicate established human lymphoma cells in a severe combined immunodeficient (SCID) mouse xenograft model in vivo. The current study was aimed at increasing the sensitivity of leukemia and lymphoma cells to CIK cells. In particular, the authors wanted to target CIK cells to leukemia and lymphoma cells via reverse antibody-dependent cellular cytotoxicity. Binding of an anti-CD3 monoclonal antibody to CIK cell cultures derived from patients with lymphoma was shown using flow cytometric analysis. For the target side, several B-cell lines were found to express CD19 on the cell surface. There was an impressive increase in sensitivity to CIK-mediated lysis of various lymphoma and leukemia cell lines by preincubation of the targets with a monoclonal antibody against CD3. This increase could be partially blocked by preincubation with anti-CD16 (Fc receptor III) and anti-CD32 (Fc receptor II) antibodies. These data suggest that the increase in cytotoxic activity is caused by Fc receptor-mediated antibody binding. Cytotoxic activity could be further increased by adding an anti-CD28 antibody in addition to anti-CD3. Finally, there was a further increase in sensitivity to CIK-mediated lysis of CD19+ malignant cells using the bispecific OKT3xHD37 antibody with specificity against CD3 and CD19. Interestingly, preincubation of malignant cells with an anti-CD3 monoclonal antibody followed by addition of the bispecific OKT3xHD37 antibody led to a further increase of cytotoxic sensitivity compared with the addition of the bispecific antibody alone. In conclusion, these data suggest that cytotoxic activity of immunologic effector cells can be increased not only by using the bispecific antibody OKT3xHD37 in vitro but also by preincubation of CD19+ leukemia and lymphoma cells with a monoclonal antibody against CD3. In addition, the immunostimulatory effect of the bispecific antibody OKT3xHD37 can be further increased by adding a monoclonal antibody against CD3. PMID- 10838660 TI - Granulocyte-macrophage colony-stimulating factor, interleukin-2, and interleukin 12 synergize with calcium ionophore to enhance dendritic cell function. AB - The authors previously showed that monocytes treated with calcium ionophore (CI) acquire characteristics of mature dendritic cells (DC) in part through a calcineurin-dependent pathway. In this study, the authors evaluated the ability of granulocyte-macrophage colony stimulating factor (GM-CSF), interleukin-2 (IL 2), and interleukin-12 (IL-12) alone or in combination with CI to induce DC characteristics in peripheral blood monocytes. Monocytes obtained by leukapheresis and countercurrent centrifugal elutriation were cultured with calcium, cytokines, or both, profiled by flow cytometry, and assessed for antigen uptake and sensitization of autologous CD8+ T cells to antigen. Monocytes treated with the combination of GM-CSF, IL-2, and IL-12 resulted in immunophenotypic and antigen uptake profiles typical of immature DC, including loss of surface CD14 expression, de novo low-level expression of B7.1, negligible CD83 expression, marked enhancement of CD40 and ICAM-1, and high major histocompatibility complex class I and II levels. A high level of antigen uptake by macro-pinocytosis was observed. In contrast, CI treatment significantly up-regulates B7.1, B7.2, CD40, CD54, and CD83 and substantially down-regulates CD14 and macro-pinocytosis, a profile consistent with mature DC. Many CI-induced modulations, but none resulting from cytokine treatment alone, were inhibited by the calcineurin phosphatase inhibitor cyclosporin A. Compared with monocytes treated with CI alone, combined treatment of monocytes with GM-CSF, IL-2, IL-12, and CI augmented B7.1 and CD83 expression and enhanced sensitization of autologous CD8+ T cells to melanoma-antigen-derived peptides. These results suggest that several independent pathways of DC activation can cooperatively enhance the function of monocyte derived DC. PMID- 10838661 TI - Intensified regression of colon cancer liver metastases in mice treated with irinotecan and the immunomodulator JBT 3002. AB - The authors recently reported that tumoricidal activation of macrophages by a new synthetic bacterial lipopeptide, JBT 3002, can augment chemotherapy-mediated tumor-cell killing. The aim of this study was to identify the mechanism responsible for the destruction of metastatic cells. Three daily oral doses of JBT 3002 before once-weekly intraperitoneal injections of 100 mg/kg irinotecan for 3 weeks significantly increased the eradication of established CT-26 murine colon cancer liver metastases compared with treatment with irinotecan alone. Immunohistochemical analyses revealed that the hepatic metastases in mice given combination therapy contained infiltrating CD8+ lymphocytes and a dense infiltrate of macrophages expressing both inducible nitric oxide synthase (iNOS) and interleukin-15. In vitro treatment of peritoneal macrophages with JBT 3002 plus interferon-gamma induced the expression of iNOS and the production of nitric oxide. In the presence of a low (subtoxic) dose of irinotecan, these activated macrophages produced significant lysis of CT-26 cells. The high level of cytotoxicity was inhibited by the specific inducible nitric oxide synthase inhibitor, NG-methyl-L-arginine. In contrast, irinotecan-mediated lysis of normal intestinal epithelial IEC-6 cells was not increased by activated macrophages. Scanning electron microscopy revealed that activated macrophages bound to CT-26 tumor cells but not to normal IEC-6 cells, confirming that nitric oxide-mediated cytotoxicity is specific for tumor cells. Collectively, the results suggest that augmentation of the therapeutic efficacy of irinotecan against colon cancer liver metastases by immunomodulation with JBT 3002 may be associated with elevated inducible nitric oxide synthase and endogenous interleukin-15 in tumor infiltrating macrophages. PMID- 10838662 TI - Dynamics of tumor cell killing by human T lymphocytes armed with an anti carcinoembryonic antigen chimeric immunoglobulin T-cell receptor. AB - Chimeric immunoglobulin T-cell receptors (IgTCR) join the antigen-binding portion of an antibody to one of the signaling chains of the TCR. A previous report described the construction and functional testing of an IgTCR gene directed against the carcinoembryonic tumor antigen (CEA). These preclinical studies showed the proper assembly and cell surface expression of anti-CEA IgTCR molecules, specific target antigen binding, and activation of T-cell effector functions. Although IgTCR-modified T cells function well in vitro, therapeutic applications in humans may be complicated by various factors, such as the availability of appropriate T-cell cytokines, high systemic levels of antagonistic soluble CEA, and antigenic diversity in tumor cell populations. The current study analyzes tumor cell killing by IgTCR-modified human T cells under conditions that more closely model those that may be encountered in persons with cancer. This analysis shows that 1) depriving IgTCR-modified T cells of interleukin-2 does not diminish anti-CEA cytotoxic T lymphocyte activity, but does eliminate killing by lymphokine-activated killer cells; 2) high levels of soluble CEA do not significantly inhibit tumor cell killing even when approximately 80% of the chimeric receptors are blocked; and 3) CEA+ tumor cells that can down-regulate cell surface CEA evade immune destruction by IgTCR modified T cells. These results have important implications for application strategies and protocol design considerations for early clinical testing of IgTCR anti-tumor therapies. PMID- 10838663 TI - Anti-tumor vaccination in heterozygous congenic F1 mice: presentation of tumor associated antigen by the two parental class I alleles. AB - Peptide vaccination of homozygous mice against syngeneic tumors using single major histocompatibility complex (MHC) class I-restricted cytotoxic T lymphocyte (CTL) epitopes elicits effective immune responses against metastatic growth. So far, single-peptide vaccination of patients against their autologous tumors seems to elicit less satisfactory results. In this study, the authors tried to determine whether effective anti-metastatic immunity requires the presentation of peptides restricted by the two parental class I major histocompatibility complex alleles in heterozygous hosts. The immune response against the H-2b-derived 3LL Lewis lung carcinoma was evaluated in heterozygous recombinant congenic F1 mice (Kk x K(b)) and (Kd x K(b)). Vaccination of such heterozygous animals with dendritic cells expressing the two parental H-2K alleles, pulsed with total tumor extract, elicited a potent anti-metastatic response. A comparable response was obtained after vaccination with tumor cells genetically modified to express the two class I alleles. In contrast, vaccination of the heterozygous mice with dendritic cells expressing only one of the parental F1 H-2K alleles or with tumors expressing only one H-2K allele failed to elicit effective immunity against tumor metastasis in recombinant congenic F1 mice. It appears, therefore, that to achieve effective anti-metastatic immunotherapy in heterozygous organisms, presentation of cytotoxic T lymphocyte epitopes restricted by the two parental class I alleles is required. PMID- 10838664 TI - Restoration of alloreactivity of melanoma by transduction with B7.1. AB - Melanoma cells are unusual because, unlike most epithelial tumors, constitutive expression of human leukocyte antigen (HLA) class II molecules is common. To elucidate the role of HLA class II expression in the immunopathogenesis of melanoma, the authors compared HLA class II+ melanoma cells to autologous B cells with respect to their ability to stimulate primary (naive) histoincompatible lymphocytes and T-cell clones (antigen experienced). Using primary lymphocytes (peripheral blood lymphocytes [PBLs]), melanoma cells were nonstimulatory when compared to autologous B cells. To determine whether this was caused by defective antigen processing, the authors used alloreactive T-cell clones, which require alloantigen presentation by a histocompatible stimulator cell but not costimulation. Melanoma cells stimulated the alloreactive T-cell clones in two of three clones tested, indicating that they processed and presented alloantigen. To determine whether the failure of melanoma cells to stimulate primary lymphocytes was caused by their inability to costimulate the T cells, the authors transduced the melanoma cells with B7.1 and achieved stable expression in more than 95% of the cells. The transduced cells were highly stimulatory, eliciting a 17- to 25 fold increase in proliferation by the peripheral blood lymphocytes compared with controls. Indeed, B7-expressing melanoma cells were more stimulatory than autologous B cells, which elicited an 11- to 15-fold increase compared with controls. These data indicate that melanoma cells fail to stimulate primary lymphocytes because they do not deliver costimulatory signals. Engineering HLA class II+ melanoma cells to express high levels of B7.1 may provide a way to elicit primary T-cell responses to melanoma-associated antigens. PMID- 10838665 TI - Monoclonal antibodies directed against the T-cell activation molecule CD137 (interleukin-A or 4-1BB) block human lymphocyte-mediated suppression of tumor xenografts in severe combined immunodeficient mice. AB - A monoclonal antibody specific for the human analog of the murine T-cell activation molecule 4-1BB was generated and is shown here to react selectively with activated human CD4+ and CD8+ T lymphocytes. Treatment of these T cells in a one-way mixed lymphocyte culture with the anti-h4-1BB antibody enhanced the cell proliferation of the allostimulated lymphocytes. Previous studies in the mouse have shown that treatment of tumor-bearing mice with antibodies to 4-1BB augments anti-tumor immunity that is mediated by both CD4+ and CD8+ T cells. The authors consider the possibility that a similar approach may be efficacious for human cancer immunotherapy. This question was addressed by evaluating the effect of an anti-h4-1BB monoclonal antibody on human lymphocyte-mediated suppression of a human tumor xenograft in SCID mice. Mice treated with a control antibody and co injected with the tumor and peripheral blood lymphocytes exhibited a lymphocyte dose-dependent suppression of tumor growth. In mice treated with the anti-h4-1BB antibody, the lymphocyte-mediated tumor suppression was completely eliminated and tumors grew progressively (as was observed in mice inoculated with tumors without lymphocytes). This monoclonal antibody specific for anti-h4-1BB, which augments the proliferation of allostimulated cells in vitro, blocks T-cell anti-tumor activity in vivo. These results suggest that although 4-1BB plays a role in the human peripheral blood lymphocyte-mediated suppression of tumor growth, antibodies to this molecule on human cells fail to stimulate anti-tumor activity, as was observed in tumor-bearing mice treated with an antibody to murine 4-1BB. PMID- 10838666 TI - Generation of monocyte-derived dendritic cells from patients with renal cell cancer: modulation of their functional properties after therapy with biological response modifiers (IFN-alpha plus IL-2 and IL-12). AB - The combination of interferon-alpha (IFN-alpha) plus interleukin (IL-2) has been accepted in the treatment of metastatic renal cell carcinoma (MRCC), whereas vaccines based on IL-12 or dendritic cells (DCs) are still being investigated. Here the authors analyzed 1) the feasibility to generate functional monocyte derived DCs (MDDCs) from patients treated with biological response modifiers (BRMs) who have MRCC, 2) the phenotypic modulations of these MDDCs during BRM treatment. Eight and 13 MRCC patients received IL-2 plus IFN-alpha or IL-12 immunotherapy, respectively. The adherent fraction of mononuclear cells from patients' blood drawn before, during, and after immunotherapy was incubated in clinically approved culture medium supplemented with 5% autologous serum, rhu granulocyte macrophage colony-stimulating factor, and rhuIL-4 for a week. At day 7 or 8 of culture, floating cells were examined in flow cytometric and functional assays (alloreactivity, proliferation assays in the presence of tetanus toxoid or tumor peptides, IL-12 secretion). In all patients except two, MDDCs could be generated but at a lower rate compared with healthy volunteers. Morphologic and phenotypical analyses revealed immature DCs with low levels of CD1a or CD83 expression throughout therapy with BRMs. Capacities in mixed leukocyte reactions were similar to those of healthy volunteers and stable during immunotherapy, whereas presentation of major histocompatibility complex class II tetanus toxoid peptide complexes was slightly enhanced during and after IL-12 therapy. IL-12 expression levels under IFN-gamma and CD40L stimulation were significantly lower in MDDC cultures from patients with MRCC compared with healthy volunteers. Overall, peripheral blood mononuclear cells from a cohort of 21 patients with metastatic disease who were treated with BRMs maintained their ability to differentiate into functional MDDCs with no selective quantitative or qualitative advantage. PMID- 10838667 TI - Development of a polynucleotide vaccine from melanoma antigen recognized by T cells-1 and recombinant protein from melanoma antigen recognized by T cells-1 for melanoma vaccine clinical trials. AB - MART-1, a melanoma antigen recognized by T cells-1, is a melanocyte lineage differentiation antigen expressed only in melanocytes and melanoma cells. This protein is recognized by many T-lymphocyte lines that are human leukocyte antigen (HLA)-A2 restricted and melanoma reactive. These observations have culminated in an array of clinical trials of MART-1 immunization using recombinant viruses or MART-1 immunodominant peptides. Polynucleotide immunization is a promising alternative to recombinant viral vaccines that allows delivery of the full-length cDNA encoding all potential peptide epitopes in a vector that is uncompromised by anti-viral immunity. In preparation for a phase I clinical trial of MART-1 polynucleotide immunization in patients with resected melanoma who were at significant risk for recurrence, the authors constructed a plasmid DNA encoding the MART-1 cDNA under transcriptional regulatory control of the cytomegalovirus immediate early promoter-enhancer and partially deleted intron A. This plasmid directs high-level MART-1 expression in transduced myoblasts and maturing myocytes diffusely throughout the cytoplasm. Immunization of mice with this construct by intramuscular injection elicited MART-1-specific immune responses in all animals. Previous trials of MART-1 immunization have been unable to examine the humoral immune response to MART-1 because of a lack of sufficient, highly purified protein. We have produced and purified Escherichia coli recombinant MART 1 protein using a glutathione-S-transferase fusion protein expression system. Protein staining of a sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed a band of MART-1 protein at approximately 20 kD; and Western immunoblotting with an anti-MART-1 monoclonal antibody confirmed a doublet at approximately 20 kD. These findings are consistent with previous reports using different expression systems for recombinant MART-1. This protein preparation functioned well in enzyme-linked immunosorbent assays (ELISAs) to detect anti MART-1 antibody responses in a mouse model; and a panel of healthy donor human sera showed minimal binding to ELISA plates coated with the protein, supporting its utility in monitoring human anti-MART-1 antibody responses. The glutathione-S transferase fusion method yielded approximately 200 micrograms MART-1 per 2-L bacterial culture, enough to coat 100 ELISA plates. PMID- 10838668 TI - Decreased tolerance to interleukin-2 with repeated courses of therapy in patients with metastatic melanoma or renal cell cancer. AB - High-dose interleukin-2 (IL-2) therapy has a response rate of approximately 20% in patients with metastatic melanoma and renal cell cancer. Animal models have shown that the anti-tumor effects of IL-2 are dose and schedule dependent, and one report has shown that patients with melanoma who responded to IL-2 therapy received more doses of IL-2 than did those who did not respond. The current study evaluated patients' tolerance to IL-2 over multiple courses of therapy and the factors that affected the number of doses delivered. Patients with metastatic melanoma or renal cell cancer who received at least two consecutive courses of high-dose intravenous IL-2 alone from October 1, 1985 through December 31, 1996 were evaluated. Patients served as their own controls in paired analyses. The number of doses tolerated from one course to the next and the reasons for stopping therapy were analyzed. One hundred fifty-nine patients received two or more courses of therapy during the study. The median number of doses of high-dose IL-2 decreased from course 1 (15 doses) to course 2 (12 doses) (p2 = 0.0001). Pretreatment factors were not found to significantly influence the decrease in the number of doses delivered. Only 2 of 33 separate toxic effects resulting in discontinuation of IL-2 dosing were found to be significantly different between courses. After adjusting for multiple tests of statistical significance, serum aspartate aminotransferase elevations were more likely to stop course 1 (p2 = 0.0033) and creatinine elevations were more likely to stop course 2 (p2 = 0.00007). The influence of renal toxicity was further assessed by comparing the median creatinine value at the time IL-2 dosing was discontinued. This difference was found to be significant when cycle 1 of course 1 (1.5 mg/dL) was compared with cycle 1 of course 2 (1.8 mg/dL; p2 = 0.0001). When pretreatment factors were analyzed, male sex (p2 = 0.006), a diagnosis of renal cell cancer (p2 = 0.008), previous nephrectomy (p2 = 0.001), and older age (p2 = 0.0055) were significantly associated with the development of renal toxicity that resulted in discontinuation of IL-2 therapy. Furthermore, the same four patient subsets had higher baseline creatinine values in individual univariate analyses. This study identified subsets of patients who tolerated less IL-2 with repeated courses. The decreasing tolerance to IL-2 was associated primarily with elevations in creatinine. Finding ways to ameliorate the renal toxicity seen during IL-2 therapy in this patient population may allow an increase in the number of IL-2 doses administered and potentially an increase in clinical response. PMID- 10838669 TI - Comparison of the PTSD Symptom Scale-Interview Version and the Clinician Administered PTSD scale. AB - The Clinician-Administered PTSD Scale (CAPS) is one of the most frequently used measures of posttraumatic stress disorder (PTSD). It has been shown to be a reliable and valid measure, although its psychometric properties in nonveteran populations are not well known. One problem with the CAPS is its long assessment time. The PTSD Symptom Scale--Interview Version (PSS-I) is an alternative measure of PTSD severity, requiring less assessment time than the CAPS. Preliminary studies indicate that the PSS-I is reliable and valid in civilian trauma survivors. In the present study we compared the psychometric properties of the CAPS and the PSS-I in a sample of 64 civilian trauma survivors with and without PTSD. Participants were administered the CAPS, the PSS-I, and the Structured Clinical Interview for DSM-IV (SCID) by separate interviewers, and their responses were videotaped and rated by independent clinicians. Results indicated that the CAPS and the PSS-I showed high internal consistency, with no differences between the two measures. Interrater reliability was also high for both measures, with the PSS-I yielding a slightly higher coefficient. The CAPS and the PSS-I correlated strongly with each other and with the SCID. Although the CAPS had slightly higher specificity and the PSS-I had slightly higher sensitivity to PTSD, overall the CAPS and the PSS-I performed about equally well. These results suggest that the PSS-I can be used instead of the CAPS in the assessment of PTSD, thus decreasing assessment time without sacrificing reliability or validity. PMID- 10838670 TI - Dimensional analysis of the Impact of Event Scale using structural equation modeling. AB - The dimensionality of the Impact of Event Scale (IES) was analyzed using structural equation modeling (SEM). Responses from 321 individuals (62% response) who had experienced a mass murder of seven people 8 months earlier were obtained. A model with a general factor and three subordinate specific factors- Intrusiveness, Avoidance, and Sleep Disturbance--was developed. Scores on the original IES subscales and the SEM factors were related to scores on the General Health Questionnaire. It was concluded that the original subscales could, to a certain degree, be regarded as a reflection of negative affectivity. A more differentiated pattern emerged using the specific latent variables from the SEM analysis. PMID- 10838671 TI - Effects of posttraumatic stress and acculturation on marital functioning in Bosnian refugee couples. AB - Forty Bosnian refugee couples living in the United States completed a translated version of the PTSD Symptom Scale--Self Report, the Behavioral Acculturation Scale, the Marital Satisfaction Inventory--Revised, and a demographic questionnaire. Posttraumatic stress disorder (PTSD) symptomatology was the best predictor of marital functioning and was related negatively to acculturation. After controlling for PTSD, acculturation did not predict marital functioning. Wives' marital satisfaction was best predicted by husbands' PTSD, husbands' acculturation, and their own PTSD. Husbands' marital satisfaction was not predicted significantly by any of these variables. These findings suggest several implications for mental health professionals dealing with refugees and other traumatized populations. PMID- 10838672 TI - Coping with domestic violence: control attributions, dysphoria, and hopelessness. AB - We investigated the influence of control judgments and coping style on emotional reactions to domestic violence utilizing the framework of hopelessness theory. We assessed abuse severity, control attributions, coping, dysphoric symptoms, and hopelessness in 70 battered women recruited from 12 domestic violence agencies. Respondents reported dysphoria but not hopelessness. Increased reports of dysphoria were associated with higher levels of self-blame and avoidance coping and lower levels of problem-focused coping. Increased problem-focused coping was associated with decreased hopelessness. Perceived control over current abuse was not related to dysphoria. High expectations for control over future events were associated with decreased dysphoria. We discuss our results in terms of their application to attributional accounts of emotional reactions to battering. PMID- 10838673 TI - Olfactory identification in combat-related posttraumatic stress disorder. AB - Recent neuropsychological conceptualizations of posttraumatic stress disorder (PTSD) implicate dysfunction of the fronto-limbic system, a brain system thought to be involved in the mediation of emotion. However, few studies have examined fronto-limbic subregions, such as the orbitofrontal cortex, in PTSD. As a measure of orbitofrontal integrity, olfactory identification was assessed in 26 Vietnam War veterans with PTSD, 25 Vietnam War veterans without mental disorders, and 17 Vietnam-era, non-war-zone veterans without mental disorders. Relative to veterans without PTSD, those diagnosed with PTSD were less proficient in odor identification and verbal learning but not on other cognitive tests sensitive to dorsolateral prefrontal and mesial temporal functioning. Results bolster prior research indicating fronto-limbic dysfunction in PTSD, and suggest involvement of the orbitofrontal region. PMID- 10838674 TI - Prevalence of personality disorders among combat veterans with posttraumatic stress disorder. AB - Many combat veterans with PTSD have co-occurring symptoms of other forms of psychopathology; however, there have been limited studies examining personality disorders among this population. The few extant studies typically have assessed only two or three personality disorders or examined a small sample, resulting in an incomplete picture and scope of comorbidity. This study assessed all DSM-III-R personality disorders in 107 veterans in a specialized, inpatient unit. Using the Structured Clinical Interview for DSM-III-R Personality Disorders, 79.4% of the participants were diagnosed with at least one personality disorder: 29.9% received only one diagnosis, 21.5% had two, 15.9% had three, and 12.1% had four or more. The most frequent single diagnoses were Avoidant (47.2%), Paranoid (46.2%), Obsessive-Compulsive (28.3%), and Antisocial (15.1%) personality disorders. PMID- 10838675 TI - Outcomes of single versus multiple trauma exposure in a screening sample. AB - Most studies ignore prior trauma exposure when evaluating outcomes of target events. This study explored symptom severity associated with different types of traumatic experiences occurring alone and with multiple exposure. The Stressful Life Events Screening Questionnaire categorized 1,909 sophomore women into groups including no trauma exposure, exposure to a serious non-Criterion A event only, exposure to several unique noninterpersonal and interpersonal events, and exposure to multiple interpersonal events. Women with noninterpersonal trauma did not differ from those without trauma on the Trauma Symptom Inventory. Only interpersonal trauma and non-Criterion A events were associated with elevated symptoms; multiple-exposure participants had significantly higher symptoms than all other groups. Complex trauma histories should be accounted for, even in studies of one target event. PMID- 10838676 TI - PTSD scale of the Child Behavior Checklist: concurrent and discriminant validity with non-clinic-referred sexually abused children. AB - Several assessment instruments include measures that are purported to assess characteristics of posttraumatic stress disorder (PTSD). Although these measures are used often by researchers and clinicians, few are supported by extensive validity data. The PTSD scale of the Child Behavior Checklist (CBCL) is one that has not yet encountered significant challenges to its validity. We examine the concurrent and discriminant validity of the CBCL-PTSD scale. Participants included 63 non-clinic-referred sexually abused (SA) children, 60 non-SA psychiatric outpatient children, and 61 non-SA, non-clinic-referred schoolchildren. Results revealed questionable concurrent validity for this scale, and suggest poor discriminant validity between SA children and non-SA psychiatric outpatients. PMID- 10838677 TI - Symptom structure of PTSD following breast cancer. AB - Identification of posttraumatic stress disorder (PTSD) symptoms and diagnoses in survivors of cancer is a growing area of research, but no published data exist regarding the symptom structure of PTSD in survivors of malignant disease. Findings from investigations of the PTSD symptom structure in other trauma populations have been inconsistent and have not been concordant with the re experiencing, avoidance/numbing, and arousal symptom clusters specified in DSM IV. The present study employed confirmatory factor analysis to evaluate the extent to which the implied second-order factor structure of PTSD was replicated in a sample of 142 breast cancer survivors. PTSD symptoms were measured using the PTSD Checklist--Civilian Version (PCL-C). Fit indices reflected a moderate fit of the symptom structure implied by the DSM-IV. These findings provide some tentative support for the DSM-IV clustering of PTSD symptoms and for the validity of cancer-related PTSD. PMID- 10838678 TI - Relationship between attachment style and posttraumatic stress symptomatology among adults who report the experience of childhood abuse. AB - This study examines the relationship between adult attachment style and posttraumatic stress symptomatology in a volunteer sample of adults who reported the experience of childhood abuse. Sixty-six individuals completed measures of abuse history, attachment style, and posttraumatic stress symptomatology. Results indicated that 76% of participants endorsed one of the three insecure attachment styles (dismissing, fearful, or preoccupied). Analyses of variances revealed that those who displayed fearful and preoccupied attachment styles, which represent a negative view of the self, had the highest mean scores on posttraumatic symptoms. Correlational analyses revealed a significant positive relationship between negative view of self and posttraumatic stress symptomatology, but not between negative view of other and posttraumatic stress symptomatology. Regression analyses indicated that having a negative view of self was most highly associated with posttraumatic stress symptoms, followed by a history of physical abuse. The regression analysis further indicated that negative view of other was unrelated to posttraumatic stress symptoms. PMID- 10838680 TI - Psychosocial resource loss as a mediator of the effects of flood exposure on psychological distress and physical symptoms. AB - This study used the Conservation of Resources stress model to examine the role of psychosocial resource loss in the aftermath of Midwest flooding. Questionnaires were distributed through churches and completed by 131 adults in flood-affected communities 6 weeks and 6 months after the flood's crest. Frequent psychosocial losses included losses of routine, sense of control, sense of optimism, accomplishing goals, and time with loved ones. Path analysis revealed that psychosocial resource loss mediated the effects of flood exposure on both psychological distress and physical symptoms at 6 months post-flood. The findings suggest that interventions designed to prevent psychosocial resource loss may reduce the long-term effects of disasters. PMID- 10838679 TI - Randomized clinical trial of brief eclectic psychotherapy for police officers with posttraumatic stress disorder. AB - The authors report on a randomized, controlled clinical trial on the treatment of posttraumatic stress disorder (PTSD), comparing manualized psychotherapy to wait list control. This is the first study to evaluate Brief Eclectic Psychotherapy (BEP), which combines cognitive-behavioral and psychodynamic approaches within one treatment method. Forty-two police officers with the diagnosis of PTSD participated in the study; 22 were randomly assigned to the treatment group and 20 to the wait-list control group. Assessments of PTSD and comorbid conditions were made 1 week before treatment, after treatment session 4, upon termination of treatment (16 sessions), and at follow-up 3 months later. As expected, no significant differences between groups were observed at pretest or at session 4. At posttest and at follow-up, BEP had produced significant improvement in PTSD, in work resumption, and in some comorbid conditions. PMID- 10838681 TI - Accident cognitions and subsequent psychological trauma. AB - This study used a sample of 72 consecutive attendees to hospital following motor vehicle accidents. It aimed to assess the relationship between demographic variables, details of the accident and cognitions about the accident recorded soon afterward, and degree of psychological trauma 3 and 6 months later. Psychological trauma was assessed using the General Health Questionnaire, Impact of Event Scale, and Posttraumatic Stress Disorder Interview. A structured interview was used to gain information about demographic, accident, and accident cognition variables. Results showed that initial cognitions such as perceived threat to life, rather than demographic or accident variables, had the strongest relationships to subsequent trauma. PMID- 10838682 TI - [3D power Doppler ultrasound: new possibilities in the diagnosis and documentation of tumors of the base of the tongue]. AB - BACKGROUND: To evaluate a novel 3-D power Doppler ultrasound technique (3-D PDUS) in the diagnosis and documentation of tumors of the base of the tongue. PATIENTS AND METHODS: Twenty patients with tumors of the base of the tongue (2T1, 6T2, 6T3, and 6T4 carcinomas) prospectively underwent two-dimensional gray scale and power Doppler ultrasound and 3-D PDUS (3-Scape). All examinations were performed with a 7.5 MHz transducer and the Sonoline Elegra Advanced. Two independent observers compared the examinations regarding clear tumor margins, maximal diameter of the tumor, midline crossing, infiltration of the mylohyoid muscle, infrahyoidal tumor, contact to the lingual artery, vascularization in the tumor and in surrounding tissue, and evaluated the documentary use of the 3-D reconstruction. RESULTS: All relevant pretreatment parameters were accessible in the 3-D data; results of both observers were almost identical. An acquisition and reconstruction of the 3-D data takes only a couple of minutes. In addition to two dimensional ultrasound 3-D PDUS allows reconstruction of axial images of the base of the tongue similar to CT images and a 3-D visualization of tumor vascularization. CONCLUSIONS: 3-Scape is a novel user-friendly method that allows uninterrupted acquisition of large 3-D volumes, and digital storage and transmission of ultrasound studies without loss of information. US images can be reconstructed in any desired plane or visualized in 3-D. In the future, additional software will permit exact determination of the volume and degree of vascularization of tumors. PMID- 10838683 TI - [Results of pretherapeutic lymph node diagnosis in head and neck tumors. Clinical value of 18-FDG positron emission tomography (PET)]. AB - BACKGROUND: Histological studies demonstrate that there is a high percentage of occult nodal metastasis in head and neck malignomas. Patients with positive lymph nodes have a comparatively worse prognosis. A neck dissection is required in these cases. By demonstrating morphological abnormalities, imaging procedures like ultrasound, computer tomography (CT), and magnetic resonance imaging (MRI) can provide important initial informations about possible malignant alteration of the lymph nodes. Positron emission tomography (PET) allows functional metabolic imaging of a suspected tumor site. The aim of this study was a comparative evaluation of different diagnostic procedures with special emphasis on the value of PET in the pretherapeutic diagnosis of nodal spread in head and neck cancer. PATIENTS AND METHODS: Forty patients (28 male and 12 female) with a suspected malignoma in the head and neck region underwent clinical examination including palpation of the neck sides, ultrasound, CT, and PET to detect a nodal spread of the malignancy. Fifty neck dissections were performed in 28 patients. Lymph node biopsies were performed in the remaining patients. The results of the diagnostic procedures were compared to the histology and the clinical course of the patients. The mean follow-up period was 12.5 months. RESULTS: A nodal metastasis was verified in 35% of all cases. Sensitivity of all imaging procedures including PET was 82%. Palpation had a sensitivity of only 61%. Specificity was 85% for ultrasound, 94% for CT and palpation, and 87% for PET. PET produced false negative results in 13.4% of all cases. Inflammation was detected in these cases. The positive predictive value was marginally better for PET than for ultrasound (77% vs. 75%). It proved to be lower than the values for palpation (86%) and CT (88%). Negative predictive value was 90-91% for all imaging procedures. CONCLUSION: In the primary diagnosis of nodal alterations in the head and neck region, a PET scan has the same diagnostic value as ultrasound or CT. By imaging the metabolism of a suspected nodal metastasis, PET can help to improve the assessment of regions with uncertain anatomic features. To avoid false positive results, acute and chronic inflammatory alterations have to be ruled out before the PET imaging. PMID- 10838684 TI - [Diagnosis of recurrences of head and neck carcinoma with the tumor marker SSC antigen]. AB - BACKGROUND: The follow-up of squamous cell carcinomas of the head and neck is often challenging. Due to tissue alteration and anatomic changes after primary treatment or submucosal tumor growth, recurrences are sometimes detected very late. The tumor marker SCC-Antigen (SCC-Ag) may provide additional information for early detection of such tumor recurrences. PATIENTS: Serum levels of SCC-Ag in 578 patients with primary squamous cell carcinomas of the head and neck were assayed by SCC-RIA and IMx-SCC before treatment and every 2-3 months during follow-up. During the observation period of 30-84 (mean 50) months, 179 recurrences were verified by histologic examination. RESULTS: Seventy-seven patients (43%) with tumor recurrence showed elevated serum levels of SCC-Ag (< 2.0 ng/ml). Fifty-eight (32%) of them exhibited elevated levels of SCC-Ag up to 11 months (mean 6.1 months) prior to histopathologic diagnosis. This mainly became evident in 48 (83%) patients whose SCC-Ag serum levels were elevated before treatment. CONCLUSION: Use of SCC-Antigen in head and neck tumors follow up can provide early evidence of almost one third of all recurrences of squamous cell carcinomas of the head and neck. For clinical purposes, we recommend an initial analysis of the SCC-Ag serum level in every patient with primary squamous cell carcinoma of the head and neck. The SCC levels of all SCC positive patients should be closely monitored. Elevated SCC should be regarded as a potential early sign for recurrence and therefore indicates the need for intensified follow-up. Depending on the individual situation this should include ultrasonography, CT, MRI and especially frequent endoscopy in general anesthesia with multiple biopsies of suspicious regions. PMID- 10838685 TI - [Locally recurring extramedullary plasmacytoma of the upper aerodigestive tract]. AB - BACKGROUND: Extramedullary plasmacytomas (EMP) are plasma cell tumors in which by definition the primary tumor is extramedullary. Most of them are found in the upper aerodigestive tract. PATIENTS: In this study we describe 3 patients with EMP. The first case is a locally recurrent EMP with recurrent involvement of cervical lymph nodes. The first manifestation of EMP was 31 years ago. Hence, this is one of the longest clinical courses of recurrent EMP ever described in the literature. Case 2 is a locally aggressive recurrent EMP. Case 3 is a localized solitary EMP that could be successfully treated by surgery alone. RESULTS AND CONCLUSIONS: Different classifications of EMP described in the literature are discussed. If these classifications are applied to our cases neither the tumor stage nor the histological picture allow definite conclusions about the prognoses to be drawn. Metastases in regional lymph nodes do not necessarily mean a worse prognosis. Overall, compared to MM with a ten year survival rate of 18% the prognosis is more favorable in EMP with a ten year survival rate of 50%. After a generalized plasma cell neoplasia has been excluded EMP in the head and neck should be treated like a locally aggressive and potentially metastatic tumor. From our experiences we recommend a primary surgical therapy followed by radiation therapy if necessary. PMID- 10838686 TI - [Experience with endonasal endoscopic surgery of inverted papilloma of the nose and paranasal sinuses]. AB - BACKGROUND: The inverted papilloma of the nasal cavity and the paranasal sinuses is a benign but locally aggressive neoplasm with a high recurrence rate and an unknown risk of malign transformation. This unsteadiness of biological behaviour requires a permanent control of the outcome of the available surgical treatments to ensure the utmost reliability for the patients. METHODS: In our investigation we analysed the surgical results in 54 patients with inverted papilloma of the last 30 years with an average followup of 55 months. 25 of them were endoscopically treated. The other group of 29 patients was treated by traditional surgical techniques using an extra-nasal approach. RESULTS: Using the endonasal endoscopic technique we observed a recurrence rate of 48% whereas the other group treated by an extra-nasal approach reached a recurrence rate of only 24% and did not show any multiple recurrences. A malign transformation was found in two patients (< 5%) within the first 8 months after the first resection. CONCLUSION: In consequence patients with inverted papilloma have to be informed of the different surgical techniques and their recurrence rates. Especially an endonasal endoscopic treatment of the maxillary sinus has to be carefully considered. PMID- 10838687 TI - [Diagnosis and treatment modalities in sinonasal inverted papillomas]. AB - BACKGROUND: Today, etiology and pathogenesis of sinunasal inverted papillomas (IP) remain unknown. Surgical approaches and extension of surgical resections are still controversial. PATIENTS AND METHODS: In a retrospective study we reviewed the charts of 90 patients with sinunasal IP. Using external (n = 78) or endonasal (n = 12) approaches the tumors were completely resected in all cases. 57 cases were investigated for human papilloma virus (HPV) by immunohistochemistry. RESULTS: 60% of IP extended beyond one single paranasal sinus or the nasal cavity. 7% infiltrated adjacent anatomical structures (subcutis, orbit, anterior skull base). The recurrence rate was 19% after a mean interval of 8.2 month. Associated malignancy was found in 7 cases (7.7%). HPV was detected in 9% of IP. Postoperative complaints were noticed by 27% of endonasal and 35% of trans-facial operated patients. CONCLUSIONS: Because of the advanced tumor stage in the majority of cases, usually a radical trans-facial surgical excision of IP is required. Endonasal approaches are feasible in selected cases, but surgery has to be more extensive compared to surgery for sinunasal polyposis. In the literature and in our experience IP is rarely associated with carcinoma (10% in literature) and the incidence of malignancy is not increased in recurrent IP. Using PCR, the prevalence of HPV ranges from 7 to 69% in the literature. Our immunohistochemical results support a rather low prevalence of HPV in IP. PMID- 10838688 TI - [Subcutaneous preservation of an amputated auricle. Morphological changes]. AB - BACKGROUND: Microsurgical replantation of an avulsed auricle remains a challenge in reconstructive surgery. Secondary reconstruction of a traumatic lost auricle is usually performed using a costal cartilage framework according to well documented techniques or with a prosthesis. In order to minimize donor-site morbidity, various efforts can be undertaken to preserve the amputated auricle by implanting the de-epithelialized cartilage framework in a subcutaneous pocket on the surface of the mastoid. Where preservation is successful, this original cartilage could be used for reconstructive treatment. PATIENT AND RESULTS: This study describes the histologic and immunohistologic changes in a complete traumatic avulsion of the auricle with subsequent cartilage conservation for eight months within a skin pocket. Trauma, preparation and preservation were accompanied by morphologic changes that included generation of local ossification centers and infiltration of fibrous tissue. We compared the macroscopic and microscopic morphology of the amputated part to native elastic cartilage following maximal denutrition and temporary heterotopic implantation in conjunction with atypical tension and pressure properties of the retroauricular pocket. CONCLUSION: In this case, the limited success of cartilage conservation in the subcutaneous pocket required conventional auricle reconstruction with autologous costal cartilage. PMID- 10838689 TI - [History of injections. Pictures from the history of otorhinolaryngology highlighted by exhibits of the German History of Medicine Museum in Ingolstadt]. AB - BACKGROUND: Injections are part of the arsenal of all medical disciplines. In addition to this common ground, each specialty has its own particular aspects; the historical development of these are presented here with respect to otorhinolaryngology. INTRAVENOUS INJECTIONS: The first experiments with intravenous injections were carried out in 1642 by a gentleman's hunting servant in eastern Germany. Similar experiments were done in 1656 by Christopher Wren, the astronomer, mathematician, and architect in Oxford, England, and a group of scientists around the physicist Robert Boyle. These experiments were prompted by new knowledge about blood circulation provided by William Harvey in 1628. The first books on the applications of intravenous infusions in humans were published in Germany by Major 1664 (Chirurgia Infusoria) and Elsholtz 1667 (Clysmatica Nova). Bladders of animals or enema syringes were used as instruments. Because of lethal accidents the infusions soon fell from favour. Kohler in Germany in 1776 eliminated a large bolus impacted in a patient's esophagus by an intravenous infusion of tartar emetic thus inducing violent vomiting. After this crucial experiment, foreign bodies in the esophagus were the most important indication for applying intravenous injections until Killian introduced extraction by esophagoscopy in 1990. CALIBRATED SYRINGES AFTER PRAVAZ: The French surgeon C. Pravaz in Lyon in 1853 invented a small syringe, the piston of which could be driven by a screw thus allowing exact dosage. A sharp needle with a pointed trocar could be introduced into the vessel making a dissection unnessessary. Pravaz used his syringe for obliteration of arterial aneurysms by injection of ferric sesquichlorate. Pravaz's syringe initiated the invention of a great number of various calibrated syringes made of glass or metal combined with glass. SUBCUTANEOUS INJECTION AND LOCAL ANAESTHESIA: The calibrated syringes were commonly used in the treatment of syphilis by mercurialization. In otorhinolaryngology, they had and still have their primary application in local anaesthesia, which was introduced by Carl Ludwig Schleich in Berlin in 1892. PARAFFIN-INJECTIONS: Around 1900 the injection of liquid paraffin for closing defects in subcutaneous tissues came into use (Gersuny in Vienna, Delangre in Tournai). This technique was immediately applied to rhinological indications such as a saddle nose (Stein 1901). This gave rise to the invention of special syringes and modifications of paraffin with different hardness and melting points. Around the middle of this century, paraffin was abandoned for this application because of serious complications, and new substances were introduced such like teflon, silicone and collagen. The historical development of these techniques of injections is described in details with many literature citations and figures. PMID- 10838690 TI - [The interesting case No. 33. Initial manifestation of HIV-1 infection]. PMID- 10838691 TI - [Molecular cytogenetic characterization of squamous cell carcinoma of the head and neck region]. PMID- 10838692 TI - Centenarians and the genetics of longevity. PMID- 10838693 TI - Coordination of metabolic activity and stress resistance in yeast longevity. AB - The genetic analysis of longevity in yeast has revealed the importance of metabolic control and resistance to stress in aging. It has also shown that these two physiological processes are interwoven. Molecular mechanisms underlying the longevity effects of metabolic control and stress resistance, as well as genetic stability, are emerging. The yeast RAS genes play a substantial role in coordinating at least the first two of these processes. Numerous correlates can be found between the physiological processes involved in yeast aging and aging in Caenorhabditis elegans and in Drosophila, and the dietary restriction paradigm in mammals. PMID- 10838694 TI - Current issues concerning the role of oxidative stress in aging: a perspective. AB - The main tenet of the oxidative stress hypothesis of aging is that accrual of molecular oxidative damage is the principal causal factor in the senescence related loss of ability to maintain homeostasis. This hypothesis has garnered a considerable amount of supportive correlational evidence, which is now being extended experimentally in transgenic Drosophila over-expressing antioxidative defense enzymes. Some of these studies have reported extensions of life span, while others have not. Interpretation of life spans in poikilotherms is complicated by a number of factors, including the interrelationship between metabolic rate and longevity. The life spans of poikilotherms can be extended multi-fold by reducing the metabolic rate but without affecting the metabolic potential, i.e., the total amount of energy expended during life. A hypometabolic state in poikilotherms also enhances stress resistance and activities of antioxidative enzymes. It is emphasized that extension of life span without simultaneously increasing metabolic potential is of questionable biological significance. PMID- 10838695 TI - Regulation of gene expression during aging. PMID- 10838697 TI - Contributions of cell death to aging in C. elegans. PMID- 10838696 TI - Crossroads of aging in the nematode Caenorhabditis elegans. PMID- 10838698 TI - Stress response and aging in Caenorhabditis elegans. PMID- 10838699 TI - Oxidative stress and aging in Caenorhabditis elegans. PMID- 10838700 TI - Mutation accumulation in vivo and the importance of genome stability in aging and cancer. PMID- 10838701 TI - Delayed aging in Ames dwarf mice. Relationships to endocrine function and body size. PMID- 10838702 TI - Stem cells and genetics in the study of development, aging, and longevity. PMID- 10838703 TI - [Sonography in acute leg thrombosis--the lower thigh is included!]. PMID- 10838704 TI - [Intraluminal sonography in the diagnosis of gastrointestinal diseases]. AB - Endosonography is an imaging method whereby a high frequency ultrasound probe is inserted into a body cavity with or without under endoscopic control. Examination of the gastrointestinal tract is performed using special echo-endoscopes or trans endoscopic mini-probes. The gastrointestinal wall, mediastinum, pancreas, bile ducts, retroperitoneum, and other structures surrounding the gastrointestinal tract are target organs for endosonography. A detailed image of pathological processes can thus be obtained. The method can be used both for primary diagnosis of lesions and in follow-up of gastrointestinal diseases. It is accurate in local staging of cancer and in detecting small lesions. There are some limitations for optimal examination like stenoses or other factors prohibiting a precise positioning of the ultrasound transducer. The clinical importance of endo sonographic examinations must be continuously evaluated on the basis of new technical modalities and changes in therapeutic procedures. PMID- 10838705 TI - [B-flow--a new method for detecting blood flow]. AB - A new ultrasound-based blood flow detection method, B-Flow is introduced. The amplitude of scatterers in flowing blood is imaged by a subtraction mode of two to four vectors along one line. Noise reduction is gained by using digital encoded US pulses. Signal enhancement is performed by using a pulse compression method (coded excitation). Compared to Color Doppler (CDI) Imaging B-Flow has a higher time (frame rate) and better spatial resolution. Typical CDI artifacts like color blooming or aliasing are missed. B-Flow does not allow velocity measurements. B-Flow has its clinical value in imaging perfused arterial vessels, simultaneous imaging of high and low velocities and delineating plaque morphology with high resolution. PMID- 10838706 TI - [Sonographic diagnosis of thrombosis of the calf muscle veins and the risk of pulmonary embolism]. AB - AIM: To determine the frequency of deep vein thrombosis in the veins of the calf muscles. The risk of embolism in relation to the localisation of thrombosis was also evaluated. METHOD: 357 consecutive patients submitted for colour Doppler sonography of the lower extremities were prospectively examined for deep vein thrombosis (DVT). Both axial and muscular calf veins were investigated. 184 of these patients underwent additional investigation for pulmonary embolism. RESULTS: Diagnosis of DVT was made by means of colour Doppler sonography in 179 patients. Soleal veins (n = 88), peroneal veins (n = 84), the popliteal vein (n = 69), and the superficial femoral vein (n = 53) were the most common sites of thrombosis. Thrombosis of the gastrocnemial veins (n = 49) occurred less frequently. 85 patients (47% of all patients having DVT) showed isolated calf vein thrombosis, in 45 patients (25%) the gastrocnemial and/or soleal veins were the only site of thrombosis. 60% of patients with symptomatic DVT also had pulmonary embolism. The embolic frequency for isolated calf vein thrombosis and muscular calf vein thrombosis was 48% and 50%, respectively. CONCLUSION: The veins of the calf muscles are a common site of acute DVT and a source of pulmonary embolism. They should always be investigated in patients with suspected DVT of the calf, and in patients with pulmonary embolism. PMID- 10838707 TI - [Value of high-resolution ultrasound in the evaluation of finger injuries in extreme sport climbers]. AB - AIM: To determine the value of high-resolution ultrasound in the evaluation of finger injuries and changes due to strain in the fingers of extreme rock climbers. METHODS: High-frequency ultrasound was performed on 208 fingers of 52 extreme rock climbers (mean age: 29.7 yrs) and on 80 fingers of 20 healthy volunteers (mean age: 28.5 yrs). The following parameters were sonographically assessed: thickness of the pulley-system A2, distance between phalanx and tendon (PS distance), gliding ability of the flexor tendons, and the periarticular and peri-tendinous space. The examination was performed on the handling in a supinated position with extended fingers, followed by active and passive flexion of about 40 degrees. All climbers also underwent clinical examination. RESULTS: The pulley-system of climbers showed a significantly increased thickness of 0.17 (+/- 0.09) cm compared with the healthy volunteers (p < 0.001). PS-distances of up to 0.51 (+/- 0.15) cm were found only in symptomatic climbers and proved to be a sign of tendon bow-stringing. No impairment of gliding ability was seen in both groups. Tendon sheath cysts were detected in 76% (62) of symptomatic fingers of the climbers. CONCLUSIONS: Non-invasive high-resolution ultrasound examination of fingers proved to be a very helpful method for diagnosing changes due to strain as well as finger injuries in rock climbers, especially in cases where the clinical examination was difficult to perform. PMID- 10838708 TI - Age-related reference ranges for fetal foot length. AB - AIM: The aim of this study was to establish age-dependent reference ranges for fetal foot length with gestation as well as for the femur/foot length ratio, based on a mathematical growth model. METHOD: In a prospective cross-sectional study of 610 pregnancies with sonographically confirmed gestational age (< 12 weeks' gestation), fetal biometry was performed in addition to measurements of fetal foot length between 12 and 42 completed weeks of gestation. Reference ranges for the fetal foot and for the femur/foot length ratio were constructed with an overall coverage of 90% of the measured data, using a previously established method of determining reference bands for growth parameters. RESULTS: Foot length was noted to level off slightly up to 24 weeks of gestation and it continued in a nearly linear fashion thereafter. The femur/foot length ratio was associated with a mean value of approximately 1 up to 24 weeks of gestation, falling to below 1 after this time point to a value of 0.85 at 41 weeks' gestation. The comparison of the reference ranges established in this study with anatomic and sonographic reference ranges for the fetal foot demonstrates good conformity with the anatomic reference range published by Streeter as early as 1920, and a large degree of conformity with the sonographic reference range developed by Mercer and co-workers. CONCLUSION: The proposed statistical approach offers reliable reference ranges for the fetal foot and for the femur/foot-ratio as well. PMID- 10838709 TI - [Multiple circular liver foci in chronic hepatic porphyria: two sonographic case reports]. AB - Appearance of liver foci in patients with chronic hepatic porphyria has been described in literature several times since 1988. We present two impressive examples of multiple coin-like lesions in the liver of patients with till than unknown porphyria. A 61-year-old women was hospitalized with suspected liver metastases: sonography showed multiple ring-like liver foci up to 2 cm. In a 52 year-old man sonography incidentally detected multiple liver foci, in this case homogeneous hyper-echogenic. In both patients a liver tumor was excluded by biopsy, the histologic examination of the specimens only showed uncharacteristic alterations. DISCUSSION: Sonographic findings in hepatic porphyria most times are only uncharacteristic alterated. But 10% of the patients show multiple hyper echogenic hepatic foci: these lesions typically have a well marked rim, some times they even appear like a ring. In color doppler examination hepatic vessels are not affected by the lesions. The lesions themselves do not show an increased vascularisation. This allows to discriminate from other liver tumors, especially from liver metastases. Histologic findings are not pathognomonic, laboratory research is subsequently required. Strictly avoiding alcoholic drinks the presented discoveries are in principle reversible. PMID- 10838710 TI - [Polymeric chondroitin sulfate vs. monomeric glucosamine for the treatment of osteoarthritis]. AB - Two potentially chondro-protective substances are compared with each other: polymeric chondroitin sulfate and monomeric glucosamine. Chondroitin-sulfate is characterized by tropism for cartilage, i.e. it is preferentially incorporated into cartilagenous tissue. This quality could be of interest in the SPECT analysis of osteoarthritic knee joints. In contrast to the classic agents Ibuprofen and Indomethacin, the antiinflammatory action of chondroitin-sulfate is less prominent, but the drug does not show any toxic side effects. Glucosamine on the other hand is a multifunctional precursor of the glycosaminoglycan synthesis in general. It lacks the cartilage tropism of chondroitin-sulfate. Into what sort of a glycosaminoglycan glucosamine is eventually integrated, depends therefore on the nature of the cells participating in the biosynthesis. PMID- 10838711 TI - [Gastritis localization and GERD incidence in patients with Cag A(+) and Cag A(-) Helicobacter pylori infection]. AB - The relationship between Helicobacter pylori (HP) and gastroesophageal reflux disease (GERD) is very complex and still not well understood. However, many issues, even those as basic as whether HP induces or protects against GERD, are still controversial. The gastritis location could play also a role in the induction of GERD. Some investigators suggested that infection by the Cag A strain of HP seems to have a positive association with reflux esophagitis. Therefore, the aim of this study was to investigate the relationship between GERD, gastritis location, HP infection and Cag A positivity. PMID- 10838712 TI - [Some aspects of focal infections of the head and neck region]. AB - The so-called focal infections are today considered to be poly-etiologic manifestations, in which there is a summation of various aggressions. The concept of focal infection is still not exactly defined. Bacterial products, toxic or antigenic substances originating from different foci are but one of the elements susceptible of unleashing the disease. This explains why there are so many foci and so few results after their elimination. The diagnosis of chronic tonsillitis or occult sinusitis especially concerning a focal disease is sometimes difficult. The ENT-specialist is responsible for both the detection of a focus in the head and neck region and its operative treatment. The interrelationship between a focal infection and its actual secondary disease must be diagnosed by the primarily inquired doctor. The diagnosis and the treatment of focal infections of the head and neck region are an interdisciplinary problem. PMID- 10838713 TI - [Bad breath--etiological, diagnostic and therapeutic problems]. AB - Oral malodor has many etiologies and is a clinical problem for many people. This paper reviews the causes and management of oral malador. In the majority of cases the problem has been shown to originate in the oral cavity. Oral malodor, a generic descriptor term for foul smells emanating from the mouth, encompasses ozostomia, stomatodysodia, halitosis (both pathological halitosis and physiological halitosis) and fetor oris or fetor ex ore. These latter terms, in turn, denote different sources of oral malodor. All conditions that favour the retention of anaerobic, mainly gram-negative, bacteria will predispose for the development of bad breath. In addition to periodontal pockets, the most important retention site is the dorsum of the tongue with its numerous papillae. During the night and between meals the conditions are optimal for odour production. Systemic pathological states, such as diabetes mellitus, uremia and hepatic diseases, induce metabolic products that are detectable as oral smells. It is always easy to recognize halitosis, but identifying the exact cause is more complex. The clinical labelling and interpretation of different oral malodors both contribute to the diagnosis and treatment of underlying disease. Treatment is directed at the underlying cause. PMID- 10838714 TI - [Fetal cells and the pathogenesis of scleroderma--microchimerism and mechanisms of alloimmunity]. AB - Fetal cells can be detected during and after pregnancy in the maternal blood circulation. Since the majority of patients suffering from autoimmune diseases are female, and scleroderma (a systemic autoimmune disease) and chronic graft versus-host disease appear to have similar symptoms, the theory emerged that these fetal cells could have some role in the pathogenesis of scleroderma. These cells could drive an aggressive immune reaction against the tissues of the patient, wherein not only auto- but alloimmune mechanisms could participate. In this brief review we try to make a short synopsis of the most important aspects of this fascinating theory. PMID- 10838715 TI - [Early-summer meningo-encephalitis (ESME) and ESME-vaccination: status 2000]. AB - Tick-borne encephalitis (TBE) is a public health problem very well under control in Austria because of a vaccination programme using a safe, efficient and well tolerated vaccine and a carefully designed social marketing concept. The Austrian vaccine underwent another technological updating and is now marketed under a new brand name (TicoVac) on the basis of an EU registration. A second product is also available (Encepur), but some limitations of use have to be taken into account. To improve the epidemiological situation even further (only 41 hospital cases in 1999) special attention has to be given to the age group 50 years and older as this is the segment of the population where the majority of cases is observed. TBE is a growing international health problem as awareness increases and cases are identified in many European countries, even in regions where TBE so far was not diagnosed. An "International Scientific Working-group on Tick-borne encephalitis (ISW-TBE)" was established to coordinate research and public health activities. PMID- 10838716 TI - Prophylactic efficiency of 3-weekly benzathine penicillin G in rheumatic fever. AB - Benzathine penicillin G (BPG) is effective for secondary prophylaxis of rheumatic fever (RF). However, interval between injections a remains a controversial matter. In a study population of 74 patients, following the initial diagnosis of RF, 3-weekly BPG (1.2 million units) regimen was started. During the first three week period, serum penicillin concentrations were examined on the 7th, 14th and 21st days and throat culture done for group-A b hemolytic streptococcal (GABHS) infection. Ten patients (13.5%) at 21st day of injection had low serum penicillin concentration after the first BPG. GABHS was isolated in 5 patients during this period. Although two of these 5 patients had symptoms of respiratory tract infection, according to laboratory data, the other three were accepted as carriers. All 74 patients were then followed-up for rheumatic recurrence (RR) during long-term period (6 to 60 months, mean 25 +/- 5 months). There was no RR among regular (missing no more than one injection a year) group. We concluded that 3-weekly BPG regimen was satisfactory for secondary prophylaxis in RF, even though serum penicillin level was inadequate during the third week in some of the patients. PMID- 10838717 TI - Clinico-bacteriological study of neonatal septicemia in Hubli. AB - Septicemia is a leading cause of neonatal morbidity and mortality in India. In a study of 242 infants with septicemia conducted between March 1996 & June 1997 at Hubli, Karnataka, 43.39% infants had 'very early onset' sepsis (VOS), 40.08%, had 'early onset' sepsis (EOS), and 16.53% 'late onset' sepsis (LOS). 54.55% neonates had birth weight below 2000 g and 39.67% were born before 37 weeks of gestation. The cardiorespiratory signs and jaundice were the most frequent clinical features. The blood culture positivity rate was 64.87%. Klebsiella species was the commonest causative pathogen found and multidrug resistance was frequent. The overall mortality rate was 47.52% and the case fatality rate in LOS was higher than in VOS and EOS (p < 0.001). The mortality was significantly higher in neonates with lower birth weight and lower gestational age (p < 0.001). The study underlines the importance of monitoring the various features of neonatal septicemia, as well as the drug resistance of the pathogens from the nurseries. PMID- 10838718 TI - Growth hormone levels in relation to birth weight and gestational age. AB - Growth hormone levels were measured in 33 umbilical cord blood samples collected from babies born at JIPMER Hospital during April and May-1998. The study was done to evaluate the growth hormone profile in relation to birth weight and gestational age. There was statistically significant difference in the cord blood growth hormone levels between babies weighing > 2500 gms (28.1 +/- 12.83 ng/dl) and low birth weight babies (76.8 +/- 55.7 ng/dl). The difference in growth hormone levels between term babies weighing > 2500 gms and preterm babies (72.5 +/- 29.4 ng/dl) was also statistically significant. However, there was no significant difference in the cord blood growth hormone levels between term low birth weight and preterm babies. Growth hormone levels were higher in preterm babies and low birth weight babies as compared to term babies weighing > 2500 gms indicating that growth hormone has an important role to play in intrauterine growth along with other growth promoting factors. PMID- 10838719 TI - Assessment of availability and working components of school health services in Delhi. AB - A total of 204 schools of different types and geographical zones were surveyed. Principals of the selected schools were interviewed to generate data regarding availability and working components of school health during last one year. School health services were non existent in 56 (27.45%) of the total schools and in another 27%, the visits made by school health team were only three or less during last one year. The situation was much better in NDMC and MCD schools but in Govt. of Delhi group, 40 of the 82 schools did not have any school health services. In most of the schools having some form of school health services (govt. or private), all the essential areas of health appraisal viz. anthropometry, general health check-up, vision testing, ENT check-up and dental examination were covered. In 21.62% of such schools no health education activity was done by the school health team. Even in the schools having some form of school health services, referral-follow up system did not seem to be working well. In this situation health check-ups merely satisfy administrative requirements and end up with some data for storage. School health services in Delhi, like other health services, are administered through multiple agencies with somewhat overlapping areas and responsibilities. This has more disadvantages than benefits. An integration of various school health schemes in Delhi with uniform system and standards should be attempted. PMID- 10838720 TI - Structural and organizational features of school health schemes in Delhi. AB - The magnitude of work and existence of multiple agencies makes the task of providing school health services to all pupils of the State of Delhi with uniform standards, a very complex job. The present article looks at the matter from the provider's end, illustrates the structural and organizational features of various school health schemes in Delhi and tries to come out with some suggestions for improvement. This study was based on qualitative research methods. Besides the three major agencies involved i.e. Govt. of Delhi, MCD and NDMC, a host of private and other agencies were also contributing significantly in covering a total of approx. 3700 schools with around 19 lakh children. Organizational and other features of school health services varied widely according to the providing agency. There is a need for integration between various school health schemes in Delhi. Since MCD school health scheme appeared to be better organized, its features can be adopted while developing uniform standards. About the components of services, promotive inputs and follow-up of health appraisals are the areas that need to be strengthened. Proper maintenance of cumulative health records, central documentation and timely dissemination of reports are also equally important and need strengthening. PMID- 10838721 TI - Nutritional status of rural pre-school children of Haryana state. AB - The present investigation was conducted on 90 rural pre-school children (1-3 years) in summer and winter from arid (Bhiwani), semi-arid (Hisar) and wet (Kurukshetra) zones of Haryana State, India. Mean daily food intake of cereals, pulses, green leaf vegetables, other vegetables, roots and tubers, milk products, fats and oils, sugar and jaggery and fruits was found lower than their respective recommended dietary intake (RDI) in summer season whereas in winter season mean daily food intake of milk and milk products provided 6, 7, and 32 per cent more than RDI in the diets of pre-schoolers of Hisar, Bhiwani and Kurukshetra zones, respectively, Similar trend was also found in nutrient intakes. Mean height and weight of few children were found lower in Hisar and Bhiwani and higher in Kurukshetra compared to their reference values. On the basis of weight for age and height for age criteria as well as clinical examinations, majority of children were found normal in Kurukshetra. PMID- 10838722 TI - Stethoscopes and nosocomial infection. AB - Stethoscopes are an essential tool of the medical profession and can become a source of nosocomial infection. A study conducted in the Department of Pediatrics, Kasturba Medical College and Hospital showed a high carriage of methicillin resistant staphylococcus (69.76%) and multi-drug resistant Gram negative bacilli (20.93%) on regularly used stethoscopes. The antibiogram of the bacterial isolates strongly suggested these to be nosocomial strains. A verbal survey revealed that regular cleaning is not common among doctors. A policy regarding cleaning of stethoscopes with an effective disinfectant may be helpful in reducing hospital-associated infections. PMID- 10838723 TI - Optimal use of laboratory tests in pediatric endocrine practice. AB - Optimal use of the laboratory in the practice of pediatric endocrinology begins with careful consideration of the patient's history and physical findings. After a review of the differential diagnosis, an orderly plan of laboratory investigation can be devised. This paper outlines some of the more common laboratory tests used to evaluate some common pediatric endocrine disorders. The intent is to provide a concise, logical approach to choose the most efficient approach. PMID- 10838724 TI - Recent advances in the diagnosis and treatment of precocious puberty. AB - In the last two decades, the diagnosis and treatment of precocious puberty has undergone important changes. The use of supersensitive assays to determine gonadotropins and gonadal hormones has increased the sensitivity and decreased the number of blood samples required to assess the diagnosis. The introduction of gonadotropin-releasing hormone (GnRH) agonists produced a revolution in the diagnosis and treatment of this disorder. Recently, the use of long acting GnRH agonists improved the adherence of patients to medical treatment and decreased the need for uncomfortable repeated doses. The medications in the treatment of the GnRH independent causes of precocious puberty, and the important revelations in the pathophysiology of these disorders, have advanced our knowledge and management of the affected children. PMID- 10838725 TI - Neonatal endocrinology. AB - Neonatal endocrinology is a diverse topic. Several chapters could be devoted to the endocrinology of fetal transition alone. The next several pages contain a brief overview of some pertinent illnesses. It is intended not to give an absolute map in the care of these patients but to help guide the physician in tailoring an approach for each patient based on current theories and practice parameters. It could also aid in improving the physician's understanding of screening laboratories used to identify those infants at risk of preventable, treatable and potentially disastrous diseases (i.e. congenial hypothyroidism). These metabolic screens are discussed due to their efficacy in the United States. In our experience, depending on the prevalence of a specific disease a few simple procedures allow for an efficient and economic way to reach ill children in a timely fashion. Other topics included in this article were reported based on their common occurrence, the lethality of illnesses if undiagnosed or their unique treatment. In the neonate hypoglycemia, either iatrogenic or secondary to sepsis, a congenital disorder of neisidioblastosis can have severe implications on the development of the CNS if not promptly treated and prevented. Some of these disorders require an experienced endocrinologist or neonatologist to treat and supervise conscientiously (i.e. CAH). However, as most of us know, it is sometimes hard to find such an individual in a community based practice. Therefore, it becomes of paramount value that each of us pays attention to the treatment of these illnesses for the sake of the children we care for. PMID- 10838726 TI - Linear catch-up growth. AB - Chronic diseases and malnutrition cause growth failure in childhood and adolescence. Correction of the cause of a growth deficiency is usually followed by catch-up growth. Capacity to catch-up is not only variable in different phases of growth, it is also different in different diseases and among different individuals suffering from the same disease. Catch-up growth is of three types. In type 1 catch-up growth, height deficit is swiftly eliminated after the growth restriction ceases. In type 2, after growth restriction ceases growth continues for longer than usual and growth arrest is compensated. Type 3 is a mixture of type 1 and type 2. Repeated episodes of growth inhibitory conditions result in lower catch-up rates in the subsequent periods. Although the exact mechanism regulating catch-up growth still remains a mystery, monitoring catch-up growth remains an important measure of the efficacy of the therapy and therefore of immense clinical importance. PMID- 10838727 TI - Goldenhar syndrome with rare associations. AB - Goldenhar syndrome is a malformation complex involving the structures arising from first and second branchial arches, the first pharyngeal pouch, first branchial cleft and primordia of the temporal bone. Though the syndrome itself is not very rare, the presence of polydactyly and hydrocephalus, which are rare associations, prompted us to report this case. PMID- 10838728 TI - Orbital granulocytic sarcoma in acute myelogenous leukemia. AB - A five year old boy presented with progressively increasing proptosis of the left eye. CT scan showed bilateral extra-conal homogeneously enhancing soft tissue masses, larger on the left side. A possibility of granulocytic sarcoma of the orbit was considered. Diagnosis was confirmed by peripheral smear and bone marrow aspiration. Patient responded to chemotherapy. PMID- 10838729 TI - Posterior mediastinal teratomas. AB - Posterior mediastinal teratomas are rare. We report this entity in a five year old boy. A review of literature of eighteen cases is presented. PMID- 10838730 TI - Infantile hookworm disease. PMID- 10838731 TI - Correlation of serum and CSF magnesium levels of normal newborns with maternal serum magnesium. PMID- 10838732 TI - [Determinants of health and health policy. Part 2. Classification and effectiveness of the determinants]. AB - Determinants of the health and disease are factors of the human biology, environment, system of health and the human behaviour. For the clinical practice not only their identification, but also their effectiveness becomes important. Modern epidemiological gnoseology serves to this purpose using methods both on the individual and the population levels. At the same time, all indexes of the population's health status respond sensitively to two factors of the social sphere: Differences in the economical standard of the population, classified according the social status or family income, and on the psychosocial markers of the population groups. Relation between poverty and diseases is apparent, while relation between the wealth and health is in the modern societies less obvious. Complicated and comparatively fast sociopolitical, cultural, and psychological changes brought about the feeling that poverty and wealth are not taken absolutely (though differences among the two has risen) but more relatively. People appreciate more the quality of life, which is based more on the psycho socio-cultural "capital", then on purely materialistic values. PMID- 10838733 TI - [Endocrinology 1998-1999]. AB - In the cascade hormone--second messenger--cellular G-proteins (GTP binding proteins), impairment can occur also at the last step: Mutant G-proteins may amplify the response (e.g. hypophyseal and thyroid adenomas) or reduce it (pseudohypoparathyreosis, testitoxicosis). Other new group of diseases appears to be anexinopathy: Among anexins belong also lippocortins and impairments occur in the hemocoagulation. "Reverse endocrinology" is a process description when the recognition of receptor (called an "orphan receptor") comes earlier than that of the hormone: Such receptors are known for several steroid hormones, retinoids and eicosanoids and it appears they are important also in the metabolism of cholesterol. A single antigen--glutamic acid decarboxylase (GAD), can cause autoimmune disease as the immuno-dependent diabetes (IDDM). Treatment of the skin T-cell lymphoma by some retinoids can result in hypothyroidism. Retrotransposones are example of the human genome modification with yet unknown clinical manifestations. Hepatocytal growth factor reveals to be the hope for treatment of cirrhosis. Search for effective peroral insulin substitutes is at present based on testing of various metabolites of fungi. Antibodies against TNF (tumor necrosis factor) become tested as "anti-cytokine therapy" in patients with rheumatoid arthritis. Some other suggestions for new ways of treatment is also listed, including the intranasal administration of estradiol. PMID- 10838734 TI - [Thrombosis in childhood--etiologic role of congenital thrombophilic conditions]. AB - BACKGROUND: Increasing frequency of thrombosis in podiatry brings about high morbidity and mortality. From published sets of clinical cases with thromboembolic complications can be concluded, that contrary to adults, origin of thrombosis in children is more frequently based on congenital thrombophilic states. The main of the work is: 1. To identify prevalence of the congenital thrombophilic states in the set of patients with venous and arterial thrombosis. 2. Formulate recommendations for the laboratory investigation. 3. Evaluate results of the thrombosis treatment in our set of patients. METHODS AND RESULTS: Set of 24 patients of the average age 6.7 years at the time of thrombosis (16 time venous, 8 times arterial) was retrospectively investigated for the presence of the factor V-Leiden mutation, prothrombine 20210A mutation, deficiency of C and S protein, and antithrombin III. Presence of acquired risk factors was also evaluated. Congenital thrombophilic state was identified in 5 patients (31.2%) with venous thrombosis and in 1 patient (12.5%) with arterial thrombosis. Mutation of the factor V-Leiden was found most frequently. It was identified at 3 patients (18.7%) with venous thrombosis and 1 patient (12.5%) with arterial thrombosis. The central venous catheter was the most frequent acquired risk of thrombosis (50%). In 1 patient with venous thrombosis and in 4 patients with arterial thrombosis no acquired or congenital risks of thrombosis were identified. Results of treatment confirmed beneficial effects of heparinisation and subsequent wafarinszation for the period of increased risk of thrombosis. Systemic thrombolysis was done 3 times without complications. CONCLUSION: Congenital thrombophilic states play significant role in the manifestations of thromboses in children. In majority of children with manifesting thrombosis at least one risk factor was identified. Cerebral infarcts in infants remain largely unrevealed. PMID- 10838735 TI - [Development of smoking habits in the population of the Czech Republic from 1985 to 1997/98]. AB - BACKGROUND: Tobacco smoking belongs to high risk factors for the circulation diseases. Aim of the present study is to identify and describe smoking habits of the population in nine districts in Czech republic in years 1997/98 and in six of these districts to analyze smoking trends during the period of 1985-1997/98. METHODS AND RESULTS: Information on the smoking habits were collected in years 1985, 1988, and 1992 in six districts which took part in the international project WHO MONICA. In 1997/98 data collection was extended into three other districts. New randomly selected samples of 1% of the population were explored each time. 5293 males and 5610 females 25-64 years old were questioned during a controlled talk with a health-officer. In 1997/98 in nine districts the prevalence of actual smokers was 38%, that of former smokers was 24% and 38% of non-smokers. The group of females consisted of 27% of actual smokers, 10% of former smokers, and 63% of non-smokers. The average daily consumption was 16.4% (+/- 8.6) cigarettes per day in males and 11.3 (+/- 7.0) cigarettes per day in females. In 1985-1997/98 the smoking prevalence of males aged 25-64 years decreased in six districts from 49% to 37% (p < 0.001). No changes were detected in females of the same age group (28% in 1985, 26% in 1997/98). Decreasing tendency was observed in both males and females up to 45 years old, in males also in the age group 55-64 years. In females older than 45 years the smoking prevalence increased. Significant changes in the smoking prevalence were found when samples were analyzed according to the education level. In males with basic education and among skilled workers the smoking prevalence decreased in years 1985-1997 from 53% to 42% (p < 0.05), among males with secondary education smoking prevalence decreased from 45% to 33% (p < 0.01) and in graduates from 34% to 23% (p < 0.01). In females with basic education the smoking prevalence increased from 25% to 31% (p < 0.05), among females with secondary education it decreased from 34% to 21% (p < 0.001), in graduate females it decreased from 31% to 18% (p < 0.05). CONCLUSIONS: Present situation and 13 years long development of smoking habits differs in males and females. Higher prevalence was found in males as well as the daily consumption of cigarettes. Since 1985 the male smoking prevalence has decreased in age groups 25-64 years and in age and education level subgroups. Only partial decrease of the female smoking prevalence was observed in age group till 45 years and in subgroups with secondary education and graduates. Significant increase in the smoking prevalence among females with basic education as well as the finding that women do not quit smoking with increasing age my become an important information for preventive programmes. PMID- 10838737 TI - [Fatal wounding of the journalist and writer, Rudolf Tesnohlidek in 1928 in the light of new findings]. AB - Presented paper describes the tragic end of the writer Rudolf Tesnohlidek, the author of the well-known and congenial masterpiece Liska Bystrouska. In archives of the Brno municipal clinics new documents have been found which may bring a new light on the circumstances of his tragic death. PMID- 10838736 TI - [Selective photodynamic destruction of leukemic cells]. AB - BACKGROUND: Residual leukemic cells if present in autologous bone marrow grafts or CD34+ concentrates obtained from peripheral blood may increase the risk of relapse after autotransplantation. We are presenting the employment of a new method which was introduced into the photodynamic therapy, namely enhancement of synthesis of the photosensitizing compound, protoporphyrin IX, in cancer cells, following application of its metabolic precursor, 5-aminolevulinic acid, for the specific destruction of leukemic cells. METHODS AND RESULTS: By determining cell viability using tetrazolium salt reduction (MTT), by flow cytometrypropidium iodide assay and by determining cell proliferation using bromodeoxyuridine incorporation we studied the effect of photodynamic therapy based on the application of 5-aminolevulinic acid on the cells of leukemic cell lines HL60 (human promyelocytic leukemia), HEL (erythroleukemia), DAUDI (B-cell leukemia), JURKAT (T-cell lymphoma), blast cells of patients with acute myelogenous leukemia as well as on normal lymphocytes and normal human bone marrow progenitors. In in vitro experiments photodynamic therapy based on an administration of 5 aminolevulinic acid (1 mM, 4 h, 18 J/cm2) lowered the number of viable leukemic cells by over 2 orders (with the exception of HEL cells) and eliminated blast cells in mononuclear cell preparations of six out of seven patients with acute myelogenous leukemia. On the other hand the viability of normal resting lymphocytes was little affected by photodynamic therapy (number of necrotic cells increased from 6 to 11%) and also the clonogenic activity of the progenitor cells of normal bone marrows did not decrease substantially (CFU-GM to 60% and BFU-E to 55% of the original activity). CONCLUSIONS: Photodynamic therapy based on the application of 5-aminolevulinic acid is a perspective method for the specific destruction of leukemic cells in autologous transplants. PMID- 10838738 TI - [Determinants of health and health policies. Part I. Causality in medicine and its modeling]. AB - Causality is the relation between the antecedent and consequence. Association between those two notions represents causal determinacy allowing understanding the subject, and the theory of probability, which deals with supposed contradiction of the chance and necessity. Understanding how health can be impaired enables to precede the disease, to change its natural history and to cure it. Causality in medicine is based on the conception of complex action of biological, psychological and social factors, which may have either positive or negative effects on our health. Empirical data forming basis for the search of aetiology represent a conglomerate of causal and indifferent elements. To identify them, various models are employed (black box, Rubik's cube, Chinese box, model of multiple accidental phenomena and others). Appropriate epidemiological methods enable not only to determine the preference, but also to signalize fallacies of looking for the origin of disease. Permanent problem of the medicine reveals the existence of the objective accident and the uncertainty in the professional decision. PMID- 10838739 TI - [Enzymatic and nonenzymatic linking elements, their development and significance in physiologic, pathologic and gerontologic changes in the body]. AB - Between the aggregated and in tissues properly deposited collagen molecules crosslinking elements, e.g. pyridinoline and deoxy-pyridinoline, are enzymatically formed. These triple-functional crosslinks are located at specific sites of the collagen chains, bind covalently its molecules, and contribute to the stability of collagen structure. Analogical quadruple-functional crosslinks are formed in the elastin, i.e. desmosine and isodesmosine, which have essential physiological functions in the body. When either collagen or elastin becomes resorbed, all the crossling elements are released into blood, and they concentrate in urine. Assessment of these elements in the body fluids can be used as an indicator of the disintegration kinetics of both tissues. Practically in all long-living tissues of the organism, a cascade of non-enzymatic chemical reactions between reducing sugars and free amino-groups generates the so-called AGE-derivatives. This family of substances, represented by pentosidine, acts in the body exclusively negatively. Their determination in tissues and in body fluids represents important markers in the clinical investigation of various diseases. The possible mechanisms of the enzymatically and non-enzymatically formed crosslinks are discussed, and the relation of AGE-derivatives to various diagnostical findings is mentioned. PMID- 10838740 TI - [Autonomic neuropathy in patients with type 2 diabetes and microalbuminuria]. AB - BACKGROUND: Microalbuminuria (MAU) represents in patients with type 2 diabetes a risk factor for total and cardiovascular mortality and morbidity, whose principal pathogenic mechanism is the development of atherosclerosis. Other factors may also participate, e.g., cardiovascular vegetative neuropathy, which is supposed to be an independent risk factor. The aim of the study was the analysis of the cardiovascular autonomic regulations in patients with type 2 diabetes and microalbuminuria. METHODS AND RESULTS: 16 patients with type 2 diabetes and microalbuminuria and 23 healthy controls were included in the study. Heart rate variability was tested (during short-term recording at rest, deep breathing, orthostasis and Valsalva manoeuvre) and spectral analysis of telemetric records of heart rate in three positions (lying--standing--lying) was employed. In the group of patients with type 2 diabetes and MAU, in comparison with patients without MAU and controls, significant differences in heart rate variability during deep breathing were found. In comparison to controls, differences were found also during the Valsalve manoeuvre. In parameters of reaction of the heart rate to orthostasis, both groups of diabetic patients differed from controls. When comparing patients with MAU and controls significant differences were also found in spectral analysis of the heart rate variability, namely in total spectral power and the power of the low frequency band in both recumbent positions. In the same parameters, significant differences were found also between patients with and without MAU. The later were not different from the controls. CONCLUSIONS: The presented results indicate the existence of a significant impairment of the autonomic nervous system in patients with type 2 diabetes and microalbuminuria. This fact may contribute to the higher cardiovascular risk in this group of diabetic patients. PMID- 10838741 TI - [Withdrawal of prednisone from a triple combination of immunosuppressive agents after kidney transplantation]. AB - BACKGROUND: Triple-drug immunosuppression may not be necessary in a majority of stabilized patients over 1 year after kidney transplantation. In contrary steroid withdrawal may be beneficial for the patient by elimination of side-effects. The primary aim of this study was assessment of the risk of rejection after the prednisone withdrawal. METHODS AND RESULTS: 88 patients 1 year after the first renal transplantation with stable graft function and serum creatinine < 160 mumol/l treated with cyclosporine-A, azathioprine and prednisone were randomized into group A (n = 46) with a prednisone withdrawal and group B (n = 42) on triple drug therapy without change. At the time of randomization, fine-needle biopsy was carried out in all of the patients. In group A, the dose of prednisone was gradually reduced to zero in the course of six months and the patients were followed up for the next 12 months. In the group B, patients on triple-drug therapy were followed for the corresponding period of time. 3 patients (6.6%) in group A, and 2 (4.8%) in group B experienced rejection (NS). Mean values of serum creatinine were in the course of follow-up in both groups without any statistical difference. Suspect immunological activity or proved immunological activity in aspiration biopsy was present in 4 patients in each group, but one of them rejected the graft. In comparison with group B, a significant decrease of cholesterol and leukocytes was observed in group A. Prednisone withdrawal had no influence on hypertension and triglyceride. CONCLUSIONS: Gradual withdrawal of steroids is not associated with higher risks of rejection and has a beneficial effect on cholesterol levels. Aspiration biopsy was of no use for the prediction of rejection. PMID- 10838742 TI - [Latent autoimmune (type 1) diabetes mellitus in patients originally classified as type 2. Divergence of etiologic markers]. AB - BACKGROUND: To assess the prevalence of markers of autoimmune insulitis (AII) in patients classified originally as having Type-2 diabetes mellitus (Type-2 DM). 386 patients subdivided according to the BMI, C-peptide and type of treatment. METHODS AND RESULTS: Age, BMI, C-peptide, Glutamic acid decarboxylase autoantibodies (GADA), HLA-DR/,-DQ alleles. Prevalence of GADA varied from < 5% in obese patients with normal/increased C-peptide to > 30% in non-obese patients with low C-peptide. In majority of GADA positive patients, the Type-1 DM high risk HLA-DRB1*, HLA-DQB1* alleles have been found. Among them HLA-DRB1*0302 and HLA-DRB1*0201 were more frequent than HLA-DRB1*040x and HL:A-DQB1*0302. CONCLUSIONS: Significant fraction of patients classified initially as Type-2 DM may have in fact Type-1 DM. Such patients can be recognized on the basis of assessment of serological (GADA) and immuno-genetical (HLA-DR/,-DQ alleles) markers. In some patients clinical, metabolic, immune, and immunogenetic markers may disagree. This divergence stresses multifactorial genesis of diabetes. Moreover, it can also suggest that both autoimmune insulitis and insulin resistance may coexist in parallel. PMID- 10838743 TI - [Cytologic diagnosis of pulmonary cancer in the Czech Republic]. AB - BACKGROUND: Cytological diagnostic of the lung cancer belongs to routine examinations. The aim of our study was to describe the development of the method in the Czech Republic during the last 20 years, and to evaluate the contribution of clinical cytologist and pathologists in this diagnostical method of the bronchogenic carcinoma. METHODS AND RESULTS: 61 stations of bronchology in the Czech Republic were requested (interrogatory method), evaluable responses were obtained from 58 stations (95.0%). Samples for micromorphological examination were send to laboratories of clinical cytology (20) or pathology (23). Samples from 15 stations are send to both departments. From the total of 91,477 of evaluated samples 64,404 were examined at departments of clinical cytology (70.4%) and 27,073 at departments of pathology (29.6%). Results from departments of clinical cytology arrive earlier (in average after 2.3 days) than from pathologies (4.9 days). Out of 4547 micro-morphologically proved lung carcinomas, 3913 (86%) were diagnosed at departments of clinical cytology and 634 (14%) at departments of pathology. CONCLUSIONS: At present, 70% of the micromorphological diagnostics is being done at laboratories of clinical cytology, 30% at departments of pathology. 86% of diagnoses of the bronchogenic cancer verified micro-morphologically come from laboratories of clinical cytology, 14% from departments of pathology. PMID- 10838744 TI - [Anorectics and pulmonary hypertension]. AB - Anorectics (appetite-suppressant drugs) are frequently requested by patients. Their usage, however, can have serious, life-threatening side effects, such as pulmonary hypertension and valve defects. The association of anorexigen use with pulmonary hypertension was first detected at the end of the sixties. Back than, the incidence of pulmonary hypertension, diagnosed as primary, increased soon after an anorexigen, aminorex was introduced. After aminorex was recalled several years latter, the incidence of the disease returned to the usual low levels. A recent epidemiological study proved that a newer anorexigen, fenfluramine (or its stereoisomer, dexfenfluramine) considerably increases the risk of pulmonary hypertension. Currently, it is unclear how the anorectics contribute to the development of pulmonary hypertension. One possibility may be the increase in plasma serotonin concentration. Serotonin is a pulmonary vasoconstrictor in many species. However, even if this mechanism plays any role in humans, it cannot completely explain the influence of anorectics on the pulmonary circulation. The anorectics cause membrane depolarization of the pulmonary vascular smooth muscle cells by inhibiting potassium channel activity. The depolarization activates voltage-operated calcium channels, thus increasing intracellular calcium ion concentration, which is the well-known stimulus for vasoconstriction. The increase in vascular tension can be especially significant when there is a deficiency in mechanisms acting against vasoconstriction, such as endothelial production of nitric oxide (NO). Such pre-existing defects may be the reason why only a fraction of patients using anorectics actually develop pulmonary hypertension. PMID- 10838745 TI - [Maternal bonding and a healthy life style during pregnancy]. AB - Maternal bonding in pregnancy appears to be an important factor, which may influence psychical and physical development of the child both before and after the birth. One of the significant determining factors of such influence is change in the general life style of the pregnant woman, which correspond with the strength of the maternal bonding. The study brings information on the basis of an empirical research, which was done, in ninetieths at a Research Institute of the Child Health in Brno. The investigated group comprised of 481 pregnant women who were all living in Brno. Results of our research bring the information about relations between the strength of maternal bonding and occupational position of the pregnant women, intensity of working activities and the way of spending their leisure time. Beside it, out research informs about some health risk activities in pregnancy and their relations to the formation of the emotional bonding towards unborn child. PMID- 10838747 TI - [Relation between cardiovascular and gastrointestinal neuropathy in diabetics]. AB - BACKGROUND: The presence of autonomic neuropathy impairs the quality of life (orthostatic hypotension, impotence, gastroparesis) or endangers the life of diabetics (sudden death, unawareness of hypoglycemia). The purpose of the investigation was: 1. To assess the presence of the autonomic neuropathy of the cardiovascular and the gastrointestinal systems and their mutual relationship. 2. To assess the relationship of found autonomic neuropathy and the subjective symptoms which are typical to affected particular systems (cardiovascular, gastrointestinal, genitourinary, sudomotor systems and the syndrome of unawareness of hypoglycemia). METHODS AND RESULTS: The group comprised of 25 type 1 diabetic patients (12 women and 13 men) mean age 40.5 +/- 11.6 (range 21-57 years) with a mean duration of diabetes of 17.8 +/- 7.9 (range 4-35 years), treated with intensified insulin regimens. The cardiovascular autonomic neuropathy was automatically examined by the VariaPulse TF 3 computer system. Scintigraphy was used to investigate the gastric emptying time of 99mTc labelled rice. The information about the subjective symptoms we collected from the questionnaire. For statistic analysis we used Spearmen correlations and ANOVA. RESULTS: 1. A statistically significant correlation was found between the presence of the autonomic neuropathy of cardiovascular and gastrointestinal systems (r = 0.634, p < 0.0007). 2. We didn't find any relation among the cardiovascular autonomic neuropathy and the subjective symptoms of cardiovascular system, respectively the gastrointestinal neuropathy (impair gastric emptying) and the subjective symptoms of gastrointestinal system. We found a significant correlation between cardiovascular and gastrointestinal neuropathy and erectile dysfunction (r = 0.48, p < 0.0078), (r = 0.42, p < 0.0388) and with the syndrome of hypoglycemia unawareness (r = 0.49, p < 0.0057), (r = 0.52, p < 0.0075). CONCLUSIONS: The evidence of the cardiovascular autonomic neuropathy is warning signal for the affected autonomic neuropathy in other systems which are more complicated for diagnostic. The subjective symptoms don't correlate with the presence of the visceral neuropathy. PMID- 10838746 TI - [Humoral profile of patients with severe aortic stenosis]. AB - BACKGROUND: While the effect of neurohormones is often studied in congestive heart failure, their role in aortic stenosis needs to be elucidated. METHODS: 54 consecutive patients with symptomatic aortic stenosis without overt heart failure were studied at the age of 64.4 +/- 9.3 yrs with echocardiography, x-ray and catheterization. Levels of circulating atrial natriuretic factor endothelin-1, catecholamines, plasma renin activity, immunoreactive insulin and C-peptide were assessed, related to hemodynamic data and compared to those in 23 healthy controls, aged 59.2 +/- 12.8 yrs. RESULTS: Patients had significantly higher plasma levels of endothelin-1 (z-value 0.64 +/- 1.19, p = 0.019), atrial natriuretic factor (z-value 2.46 +/- 2.46, p < 0.001) and dopamine (z-value 0.91 +/- 2.33, p = 0.02). Levels of endothelin-1 and ANF positively correlated with mean (r = 0.631, p < 0.001) and wedged pulmonary artery pressures and with left atrial diameter index (r = 0.602, p < 0.001). Endothelin-1 levels correlated negatively with aortic valve area (r = -0.306, p = 0.041). No correlation was found between neurohumoral plasma concentrations and left ventricular mass index. CONCLUSIONS: In patients with symptomatic aortic stenosis without overt heart failure, elevated plasmatic levels of endothelin-1, atrial natriuretic factor and dopamine were documented. The increase of ET-1 levels is related to pulmonary hypertension and severity of the disease. Left ventricular hypertrophy is not related to neurohormonal levels. Neither circulating system renin-angiotensin nor noradrenaline are activated in these patients. PMID- 10838748 TI - [Smoking in the United States and Europe]. AB - Article reports on new findings in smoking prevalence, incidence of cancer and preventive actions against smoking in the United States and some countries in Europe. Situation in the Czech Republic is described and data from the National Cancer Registry are included. PMID- 10838749 TI - [Beta carotene in the prevention of civilization-related diseases]. AB - The objective of the article is to provide a concise overview of the conclusions of most important studies assessing the role of beta-carotene in the prevention of civilization diseases. Beta-carotene seemed to be a very promising agent in the prevention of civilization diseases in the early 1980s. However, several large-scale studies concluded that the preventive effects of beta-carotene may have been overestimated or that beta-carotene may even have harmful effects previously not anticipated. Current nutritional recommendations promote increased consumption of vegetables and fruits, but beta-carotene supplementation is not recommended. PMID- 10838750 TI - [Ethics and statistics: difficult approaches to data]. AB - The objective of the paper is to draw attention to incorrectness of various statistical approaches to research data. We critically assess the situation when first the decision on the hypothesis to be proved is done and then data are made to fit the hypothesis. We demonstrate that data cannot be satisfactorily interpreted without close attention to the manner of their collection. Finally we criticize opportunistic data torturing when researcher explores the data until a significant result is found and then devises a biologically plausible hypothesis. PMID- 10838751 TI - [Pathologic anatomy in my life and work. A greeting at the commencement of the 24th nationwide Congress of Czech Pathologists held 17 June 1999 at the Medical School of Charles University in Motol]. PMID- 10838752 TI - [T and NK cell peripheral tumors]. AB - Peripheral T lymphomas are formed by cells with CD3 antigen which contain TCR alpha/beta. A minor part of the non-tumourous population contains TCR gamma/delta. The position is similar in T cell tumours. In the tumours however TCR may be lacking .NK ("natural killer") cells are CDR3 negative, rearrangement of the gene TCR does not occur and they are TCR alpha/beta and TCR gamma/delta negative. Contrary to T cells they contain antigen CD56 ("NK-associated antigen"). In these cells and in tumours cytotoxic proteins are usually present: TIA-1, granzyme-B, perforin. There is a continuous transition between T and NK cells. The prognosis of T cell tumours cannot be assessed from the cell size. T/NK small cell lymphomas are highly aggressive tumours. The prognosis of patients is very bad. From the morphological aspect they can resemble very much non-tumorous lymphocytes. Conversely, large cell T lymphomas of the skin which are CD30 positive take a very mild course and their prognosis is favourable. The author mentions also some more recent entities of extra-nodular T lymphomas: angiocentric NK/T lymphoma of the nose (and other regions), malignant T cell lymphoma in enteropathies, T cell lymphoma resembling subcutaneous panniculitis, hepatosplenic gamma/delta T lymphoma. These tumours are rare in this country and some, e.g. T/NK lymphoma of the nose, are found usually only in certain regions of the world. They may be however encountered also in Europe, in particular in immunosuppressed subjects and patients after organ transplantation. PMID- 10838753 TI - [Biopsy of the heart valves. 872 cases]. AB - In this study, 872 heart valves surgically excised from 810 patients during a period of 5 years (1994 through 1998) were examined pathologically. There was a predominance of aortic (506 patients) versus mitral valves (246 pts.). While aortic valves came more often from men (364) than from women (142), in mitral valves the M:F ratio is 82/164. Isolated calcific aortic stenosis appeared as the most frequent valvular disease (418 pts.), with predominance of its sclerotic senile type (238 pts.). Mitral stenosis (185 pts.) remains the classical post rheumatic disease. The relative frequency of a subvalvular stenosing mitral lesion is stressed. The "pure" incompetence of both aortic (70 pts.) and mitral (56 pts.) valve was usually based on valvular myxoid degeneration. An aorto mitral disease requiring replacement of both valves (51 pts.) presented typically as a post-rheumatic lesion, however, a combination of a post-rheumatic mitral with a degenerative-sclerotic aortic valve disease may be possible. In 30 patients, the valvular replacement was performed for infective endocarditis or a post-IE lesion, mostly of the aortic valve. With the almost non-existence of acute rheumatic fever and with the increasing average age of population in this country, we may expect a long-term decline in mitral valve disease and an increase in aortic valve disease, particularly in the sclerotic type of aortic stenosis. PMID- 10838754 TI - [Fetal rhabdomyoma of the mucosa of the lip (case report)]. AB - In a 15-year-old male the foetal type of rhabdomyoma was diagnosed in the mucosa of the lower lip. Diagnosis of this relatively rare tumour prevents possible mistaking for a malignant process and an inadequate therapeutic procedure. PMID- 10838755 TI - [Angiogenesis in tumors. Part I. Its role in the determination of biological characteristics of tumors; factors controlling the onset and development of angiogenesis]. AB - The tumor stroma gains increasing attention in recent years. Angiogenesis (AG) is defined as formation of new vessels from the endothelium of the preexisting vasculature. The vascular bed does not only provide supply of nutrition factors and oxygen as well as elimination of metabolic waste, but the neo-formed vessels represent also a gate for lymphogenous and hematogenous metastatic spread of the tumor. For tumor growth beyond the size of several mm, AG must be started to form vascular supply of tumor cells. After the switch to angiogenic phenotype, the tumor starts to exhibit rapid, almost exponential growth. AG is a complex process, involving degradation of the basement membrane of preexisting vessel, proliferation of endothelial buds or solid strands towards the angiogenic stimulus, maturation of endothelial cells with formation of a luminized capillary, connection of such capillaries with creation of vascular loops, and finally formation of a functional vessel, surrounded by basement membrane and pericytes. The most intensive AG is observed at the periphery of the tumor. AG is regulated by numerous angiogenic and anti-angiogenic factors, which can be synthesized and released by tumor cells themselves, stromal elements, as well as inflammatory cells, namely macrophages. The issues of regulation of AG and importance of AG for tumor growth, progression and metastasis are discussed. PMID- 10838757 TI - [In Process Citation] PMID- 10838756 TI - [Proliferative activity in pulmonary carcinoids]. AB - AIM: On the basis of known Ki-67 dependence on tumor malignancy in some lesions, we compared this marker expression quantitatively in pulmonary typical and atypical carcinoid tumors and attempted to predict their biological behavior especially in cases associated with tumorous lymphadenopathy, satellites, and carcinoid tumorlets. METHOD: Using material from surgically treated patients, we examined 54 cases of pulmonary carcinoids divided into five groups. 1. Forty-two typical carcinoids (TC), 2. Twelve atypical carcinoids (AC) diagnosed according to modified Arrigoni's criteria (Travis et al., 1998), 3. Thirty-two TC without metastases, satellites, and tumorlets (M, S, T), 4. Eight AC without M, S, T, and 5. Fourteen TC and AC associated with M, S, T. Groups 3, 4, and 5 were formed of cases selected from group 1 and 2. The proliferate activity was evaluated by Ki 67 (MIB-1, Immunotech France, 1:25). Its nuclear labeling was counted in more than 50 HPF and calculated as a number of positive nuclei in 10 HPF. The Fisher exact test was used for statistical analysis. RESULTS: The Ki-67 nuclear expression was found in 19 (45%) out of 42 TC and in 9 (75%) out of 12 AC. In the set of TC without metastases (M, S, T), the Ki-67 positive labeling was found in 14 (44%) out of 32 cases (group III) and in six (75%) out of eight AC (group IV). In all TC and AC tumors with M, S, T (group V), the Ki-67 expression was encountered in 8 (57%) out of 14 cases. The Fisher exact test showed no significant difference between all examined groups. CONCLUSION: No statistically significant difference was found in Ki-67 expression in pulmonary typical and atypical carcinoids. It appears to be a factor which can not be used for tumor prognosis prediction or adjuvant therapy indication in surgically treated patients. PMID- 10838758 TI - [The method of DNA isolation can affect the rate of preferential amplification of alleles by the polymerase chain reaction]. AB - Today, polymerase chain reaction is a common part of approaches serving for identification of individuals in legal medicine. This method is easily practicable, however attention must be paid to the optimization of reaction conditions and to the interpretation of results. From the literature, such cases are known, in which during amplification of extremely small amount of DNA (e.g. from one cell) the polymerase chain reaction preferably amplifies only one of two in the template DNA present alleles. If the amplified fragments differ in length, the shorter one is amplified preferably, and it may be cause of false results. In the presented study, DNA from 23 stains of male blood on different fabrics was isolated by two different methods (by treatment with proteinase K and boiling and by treatment with Chelex 100). The obtained DNA samples were amplified using primers, they are complementary to the amelogenin gene sequences. The system is suitable for sex determination, because amplification of the X-chromosomal sequence provides a fragment in length of 632 bp, amplification of the Y chromosomal one a fragment in length of 443 bp. The isolation method based on proteinase K led in 17.38% of samples to the very intensive preferential amplification of the longer allele, and therefore to a false result. The isolation method based on Chelex 100 provided in all cases correct results with clearly recognizable preferential amplification of the shorter allele. The reported results accentuate the meaning of choice of the appropriate isolation method, the need of accurate PCR optimization, and the careful interpretations of its outputs. PMID- 10838759 TI - [Paraffin-embedded tissues as a source of control DNA]. AB - During elucidation of crime cases, the need of a sample of DNA of the victim of the violent attack appears very often. Regarding to the obligatory archiving of histopathological preparations from sectioned persons, the paraffin-embedded tissues are easily achievable material suitable for the given purpose. Such a modification of an isolation method was elaborated which allows the yield of the sufficient amount of DNA from one histological section, and its usefulness was tested on tissues of different types and from different persons. The method was used successfully during solving of concrete cases. PMID- 10838760 TI - [Views of forensic medicine and criminology on the pathodynamics of homicide]. AB - Trauma and violence represent the domains of forensic medical expertise. The objective finding on the victim of the homicide makes it possible to reconstruct the way and mechanism of the injury connected with it and thus to determine the cause of death. The traces of violence contain many indices pointing at specific personal characteristics of the culprit, his motivation to the crime and his state of mind at the time of the homicide. To judge the penal responsibility and the guilt of the culprit it is important for the court to have the analysis of the dynamics and of the causal background of the crime, especially the synthetic evaluation of subjective, situation, or eventually psychological factors of violence with the mortal effect. The interdisciplinary forensic-medical, forensic psychological and psychophytological approach will make it possible to provide the court with a complex forensic expertise. PMID- 10838761 TI - [Evaluation of methods of trace analysis of various opiates including hydrocodone and hydromorphone in the blood and urine using gas chromatography-mass spectrometry]. AB - A GC-MS method suitable to conduct practical trace toxicological analyses of various opiates including hydrocodone or hydromorphone is presented both in scan and selected ion monitoring modes. Silylation can be a convenient reaction to derivatize unknown analytes in toxicological samples. However, silylation of drugs with tautomeric keto/enol groups in molecular structure can cause analytical problems due to poor reproducibility of conversion degree. After using N-Methyl-N-trimethyl-silyltrifluoracetamide (MSTFA) acceptable results were achieved. PMID- 10838762 TI - [Very distal finger replantation]. AB - The paper presents the authors' experiences with very distal replantations of fingers. The results of surgical treatment in eleven cases of finger or thumb amputations at distal interphalangeal joint level or more distal are reviewed. There were 3 female and 8 male patients aged 13-45 years (mean age 25 years), with 7 amputations of the thumb and 4 amputations of other fingers. In 9 cases (82%)--8 complete amputations and 1 incomplete amputation--the replanted finger survived. In 2 failures cases, neuro-vascular flaps were used for pulp reconstruction (the authors' own modification). In 10 cases a very good functional and cosmetic result was achieved. PMID- 10838763 TI - [Results of treatment of traumatic elbow dislocation in children]. AB - Between 1987 and 1997 the authors treated 49 children with traumatic dislocation of the elbow joint. The study presents the results of treatment of 26 children, mean age of 10.7 years (ranging from 5.5 to 14.5 years) at the time of trauma. The mean follow-up time was 4.3 years (ranging from 1 to 10.5 years). In one case an anterior dislocation was observed, while in the remaining cases posterior dislocation was diagnosed. Twenty-five children were treated with closed reduction of the dislocation, followed by immobilization in a plaster cast. One case with a chronic dislocation was treated surgically. Concurrent injuries were observed in 18 (69.2) children. The most common concurrent trauma was a fracture of the medial epicondyle (44% of cases with additional trauma). The following additional fractures were also noted: fracture of the lateral epicondyle, fractures of the forearm, of the neck of the radius, of the olecranon, epiphyseolysis of the distal epiphysis of the radius with a concomitant fracture of the medial epicondyle. In 19 cases very good result was achieved. In 7 cases sequels of the dislocation were noted. These were: in 3 cases flexion contractures (5 degrees, 10 degrees, 15 degrees) of the elbow joint, in two cases periarticular ossification (in 1 case this entity was treated surgically with very good result), in 1 case an asymptomatic non-union of the medial epicondyle with a 30 degrees flexion contracture in the elbow joint, and in 1 case (treated surgically) a 90 degrees flexion contracture with a movement range up to 105 degrees with degenerative changes visible on X-ray in the joint was noted. PMID- 10838764 TI - [Principles and results of treatment of forearm shaft fractures with Rush pins stabilization]. AB - Rush pins are used for stabilization of forearm shaft fractures after surgical reduction. The pins are bent intraoperatively to conform to the natural curvature of the radius. Introduction of the pin into the radius takes place between the extensors of the index, the extensor digitus communis and the extensor pollicis longus. Safe and equally effective introduction of the pins is also possible through the olecranon when treating fractures of the proximal part of the ulna or from distal end in fractures of the distal half of the ulna. The clinical material in this paper consisted of 21 patients treated surgically with Rush pins because of fractures of the forearm. In two cases primary consolidation was not achieved. Union of the fracture did occur after periostage and iliac bone grafting. Long term follow-up of 19 patients demonstrated a full range of motion in the involved extremity in 14 cases. Limited rotation of the forearm was noted in 5 patients. PMID- 10838765 TI - [Development of the normal infantile hip joints assessed by MRI]. AB - The paper provides an overview of the time of appearance of the secondary ossification centers and closure of the growth plates of the acetabulum and proximal epiphysis of the femur: the triradiate cartilage, the acetabular roof growth cartilage, the subcapital growth cartilage, the growth cartilage of the major trochanter, the growth cartilage of the minor trochanter. The study is based upon 62 MRI scans of healthy hips in 45 patients aged 3-21. The examined hips showed no pathologic traits--neither in the MRI scan nor in X-ray investigation. In Spin Echo and Turbo Spin Echo sequential imaging all obtained slices were used, on GRADIENT ECHO: FISP 3D, FLASH 2D, and FLASH 3D FAT SAT only chosen slices were included in the study. This way the following results were obtained: the ossification center of the major trochanter appears at the age of 3 in girls and at the age of 6 in boys, while the ossification center of the minor trochanter appears at the age of 6 in both sexes. The times of complete ossification of following growth cartilages were observed: for the triradiate cartilage ossification was observed at age 12-15 in girls and 15-16 in boys; for the cartilage of the acetabular roof ossification was noted at age 12-15 in girls and 15-18 in boys; ossification in the subcapital growth cartilage occurred at age 15-17 in girls and 16-18 in boys; the major trochanter growth cartilage ossifies at age 15-16 in girls and 16-18 in boys; for the minor trochanter ossification of the growth cartilage occurs at age 14-16 in girls and at age 16 18 in boys. The secondary ossification center of the pubic bone appears at age 9 11 in girls and 13-16 in boys and the secondary ossification center of the acetabular roof appears at age 13-17 in girls and boys. This study expand our knowledge on the development of the hip joint and facilitate the assessment of hip pathology. PMID- 10838766 TI - [Inter-observer variation of the Caterall and Herring classification in Perthes disease]. AB - The reliability of the Caterall and the Herring classifications (inter-observer study) was compared using radiographs of 63 hips in 58 patients. The radiographs were reviewed by three independent observers with various clinical experience. The results were analyzed using kappa statistics and the percentage of inter observer agreement. The less experienced observers seemed to be more prone to classify the hips as more affected than they actually were. They also seemed to produce a higher agreement rate. Disagreement was most common in the Caterall groups II and III. The Herring classification appears to be more reliable, also in children older than six, which perhaps makes it more useful in deciding between conservative or operative treatment. PMID- 10838767 TI - [Clinical evaluation of the hip joint after implantation of cementless Mittelmeier type and Parhofer-Monch type hip prosthesis]. AB - Total hip replacement is the most common procedure in the treatment of severe degenerative changes in the hip joint. The authors present clinical evaluation of 129 patients (151 hips) who underwent total hip replacement using either the Mittelmeier type or the Parhofer-Monch type hip prothesis. The age of the patients ranged at the time of the operation between 17 and 74 years (average 44.9). The follow-up ranged from 24 to 143 months (average 61.2 months). The P-M prothesis was implanted in 53 hips, the P-M "Plasmapore" prothesis in 42 patients, Mittelmeier Autophor 900 prothesis in 49 patients and the Mittelmeier Autophor 900S prothesis in 7 patients. Clinical evaluation was performed according to the d'Aubigne-Postel method with the Charnley modification. The results were graded as very good in 17 cases, good in 64 cases, satisfactory in 53 cases and poor in 17 cases. Very good and good results were mainly observed after implantation of the P-M type prothesis and P-M "Plasmapore" type prothesis. Worse results were observed in patients, who at the time of the operation were aged 45 years or less and who had congenital hip dysplasia. P-M type prothesis gives better clinical results in the treatment of degenerative changes of the hip joint. PMID- 10838768 TI - [A comparison of kinesitherapy with and without continuous passive motion (CPM) after the entire allograft surgery of the knee]. AB - A prospective randomized single blind investigation after total knee arthroplasty was performed to evaluate two rehabilitation protocols (one with CPM and the other without CPM). A series of 87 consecutive total knee arthroplasties in 76 patients (72 women and 4 men) was analyzed. The average age of the patients was 64.7 years. Kinesitherapy with CPM was performed in 30 (34.5%) cases, where as in 57 (65.5%) cases kinesitherapy without CPM was performed. Early functional results according to the HSS scale were assessed on the 14th day after surgery. In the group treated with CPM there were 26.7% excellent results, 70.0% good results and 3.3 satisfactory results. The group treated without CPM there were 26.3% excellent results, 70.2% good results and 3.5% satisfactory results. No statistically significant differences were noted between the results yielded by these two rehabilitation protocols. PMID- 10838770 TI - [Results of the disk microsurgery of lumbar herniated nucleus pulposus]. AB - Results of microsurgical discectomy (MSD) carried out in 151 patients with lumbar disc herniations are presented. The advantage of microsurgical discectomy is reduction of surgical trauma, better differentiation of anatomical structures, gentler manipulation around nerve roots and the dural sac, and the possibility of a meticulous hemostasis preventing profuse bleeding. Further advantages of MSD are: possibility of direct insight in intervertebral space, limited risk of peri- and postoperative complications and infection, reduction of nursing care, favourable psychological and cosmetic effects. Also possibility of improving microsurgical technique be performing these frequent in neurosurgical practice procedures is not insignificant. Final results were evaluated 6 months after MSD using the Functional Economic Rating Scale developed by Prolo et al. Very good outcome (score 8-10) was achieved in 129 patients (85.4%), good (score 6-7) in 18 (11.9%) and unsatisfactory only in 4 (2.7%). PMID- 10838769 TI - [Retrospective evaluation of the knee function after partial resection of the fat body of the knee]. AB - A series of 64 male patients aged 16-58 years (average 37 years) underwent in the years 1992-1997 a partial resection of the Hoffa pad. The results of this procedure were evaluated. The patients underwent surgery for diagnosed tear of the medial meniscus. At arthrotomy the meniscus was found to be intact and the only visible pathology appeared to be hypertrophic Hoffa pad impinging between the articular surfaces of the joint. The authors attempted to answer the following question: is partial resection of the fat body of the knee a therapeutic procedure or an excuse to justify surgery? The results presented in this paper confirm the therapeutic usefulness of this procedure. PMID- 10838771 TI - [Lateral lumbar disk herniation]. AB - The authors present the clinical evaluation of 41 patients with extreme lateral disc herniation in the lumbar spine. This group of patients represented 3.1% of all patients operated to to discopathy of the lumbar spine at the Department of Neurosurgery of the Medical Academy of Bialystock in the years 1988-1997. Males constituted 95.1% of the study series, with an age range 41-60 years. In 92.7% of the patients extreme lateral herniation was diagnosed either at L4-L5 level or L5 S1 level. In 78.1% of the cases different types of motoric disturbances were observed, while in 65.8% radicular type disorders in superficial sensibility were noted. PMID- 10838772 TI - [Distal transfer of the greater trochanter among other surgical procedures for the treatment of hip deformity]. AB - Distal greater trochanteric transfer is a rarely used procedure for treatment of hip deformity. Surgical technique is described. Potential for changing morphology of proximal end of the femur is underlined and pitfalls indicated. The main advantage of the procedure is reconstruction of the femoral neck. It is biomechanically more sound than valgus osteotomy done frequently in similar cases. Advantages of trochanteric transfer are confirmed by other authors who after this procedure achieved clinical and radiological improvement in patients with degenerative disease of the hip. PMID- 10838773 TI - [Choice of correct stabilization in long bone fractures]. AB - The paper presents an algorithm to facilitate the choice of the correct stabilization system in fractures of long bones. The algorithm is based on fracture morphology. The indications and contractions for surgical treatment are discussed. PMID- 10838774 TI - [Morphology of a lithic lesion of the bone treated with cement filling with the addition of methotrexate]. AB - The results of histopathologic examination of a lithic bony lesion and of the surrounding soft tissues obtained from a metastatic site from 2 patients are presented. Both patients had been previously treated surgically with a 40 g bone cement filling with 2 g of methotrexate. Revision surgical intervention was necessary because of loss of stability at the fracture site in a 68 year old female after with a metastasis from breast cancer after 8 weeks and in a 72 year old male with metastasis from cancer of the prostate after 10 weeks. During revision surgery the metal plates and the cement were removed. Tissues samples from the fracture site were taken for histopathologic examination. Fibrosis of connective tissue and haemosiderin accumulation were noted in the soft tissues surrounding the fractures site. Where there was direct contact between the bone cement and bone, necrosis of bone trabeculae and granuloma like bodies were noted. Revision surgery 10 weeks after the first procedure revealed more advanced bony necrosis. No cancerous tissue was observed in any of the cases. PMID- 10838775 TI - [Diskectomy of the lumbar spine: indication and contraindications]. AB - The authors present an overview of the principles of surgical treatment of lumbar disc herniation which are currently applied at spinal surgery ward by the Orthopedic Department of the Poznan School of Medical Sciences. Most of these principles are valid in spinal surgery centres in USA and most European countries. The authors reviewed surgical technique, indications, contrindications and complications related to discectomy in the lumbar spine. Although the articles deals mainly with "classical" discectomy, other surgical techniques (microdiscectomy, percutaneous discectomy, chemonucleolysis, and endoscopy assisted discectomy) are also briefly reviewed. Careful selection of patients, meticulous analysis of radiographic findings and proper surgical technique yield good results which allow most patients to improve their life conditions and lead a pain-free existence. PMID- 10838776 TI - [Characteristics of coagulase-negative staphylococci isolated from hemocultures]. AB - The authors investigated 352 strains of coagulase-negative staphylococci isolated from hemo-cultures of patients with assumed bacteraemia who were hospitalized in standard hospital departments and intensive care units. The strains were identified using STAPHYtest 16 and 14 different species and subspecies of coagulase-negative staphylococci were found. Most frequent were strains of Staphylococcus epidermidis (53%), strains of S. hominis subsp. hominis (22%) and S. haemolyticus (13%). In the strains the production of slime and delta haemolysin was investigated. The formation of these two important factors of virulence was recorded in 95% strains (incl. major slime production in 13.4%), and 50% strains resp. The resistance to 13 antibiotics was also assessed. A total of 91% strains were resistant to penicillin, 63% to oxacillin and amoxicillin with clavulanic acid, 67% to gentamicin and 66% to erythromycin. In the group 15 strains of a recently described subspecies S. hominis subsp. novobiosepticus was identified. These isolates displayed most frequently resistance: they were resistant in 100% to five antibiotics (penicillin, oxacillin, erythromycin, clindamycin and amoxicillin with clavulanic acid) and with a single exception to another three antibiotics (chloramphenicol, gentamicin and ciprofloxacin). In these strains most frequently slime as well as delta-haemolysin was found. The authors assume, consistent with previous work, that it is important to focus attention on the incidence of strains of S. hominis subsp. novobiosepticus which very probably play an important role as agents in hospital infections. The authors describe also detection of one strain of S. cohnii subsp. urealyticum which was resistant to 10 of 13 antibiotics. The authors are convinced that also classical phenotypic analysis can contribute in an effective manner to the solution of the problem of infections caused by coagulase-negative staphylococci. PMID- 10838777 TI - [Health and economic benefits of mandatory regular vaccination in the Slovak Republic. II. Poliomyelitis]. AB - The authors compared intervention strategy (vaccination) and non-intervention strategy (no vaccination). For the evaluation of health and financial effectiveness of regular polio vaccination the authors used the modelling of treatment of polio cases in 1957, and vaccination costs in 1996--the most recent year with available statistical data from the Slovak Republic. From these model years they calculated: the cost saving (11,919,106 Sk), cost-effectiveness ratio (79,944 Sk) and the net cost of vaccination (3,047,786 Sk). Simultaneously, they present a breakdown of cost-benefit analysis. PMID- 10838778 TI - [Filamentous fungal infections in man. Alphabetical overview of mycotic agents with data on their systematic classification and on the localization of individual diseases]. AB - The authors present a survey of filamentous microscopic fungi causing human mycoses. The outline is based on an alphabetical list of genera and their classification into higher taxonomic categories, i.e. orders, classes and divisions. The name of a relevant mycosis is followed by enumeration of the individual species and localizations of diseases produced by a given agent. To facilitate orientation in the system of myco-pathologically important agents, examples of classification of individual genera are presented in the text in a hierarchical arrangement of higher taxonomic categories. PMID- 10838779 TI - [Experimental Trichophyton rubrum infection in animals]. AB - The objective of the work was to induce experimental infection with the anthropophil dermatophyte Trichophyton rubrum in animals and to confirm it by clinical, mycological and histopathological examination. For preparation of the inoculum the authors used a T. rubrum culture isolated from patients suffering from dermatomycosis. The inoculum was cultivated under conditions of intensive aeration (shaking). Its density was 2.0 x 10(6)/ml of germinative spores and hyphal fragments. On the animal's back shoved and irritated by scarification (area 4 cm2) 2 ml inoculum were rubbed in. Experimental mycotic infection was induced in one of four guinea pigs and one of two domestic rabbits. The first clinical manifestations of mycotic infection were found on the 6th and 9th day after inoculation. From the focus T. rubrum was cultivated. From the unaffected hair in the close vicinity and at more remote sites numerous contaminants were isolated, other fungi as well as dermatophytes. In the histopathological material the authors found in the shed layers of the stratum corneum PAS positive septate fibres. Numerous PAS positive septate fibres were found also in the hair follicles. In the corium a mixed inflammatory infiltration was present with a predominance of histiocytes and polymorphonuclear leukocytes. No morphological changes were found after passaging the dermatophyte T. rubrum via animals. Guinea pigs and the domestic rabbit are useful animals for inducing experimental infection with the dermatophyte Trichophyton rubrum. PMID- 10838780 TI - [Use of the results of monitoring in the practice of district public health personnel]. AB - The author presents her experience with the introduction and implementation of the system of monitoring of the health status of the population and environmental factors on the territory of the Karvina district after 1989. It provides evidence that the results of monitoring need not be conceived only as systematic collection and analysis of data and evaluation of trends but also as valuable material for supervision by organs of the hygiene service. It provides also information for authorities of the state administration and self-administration, the public, teaching and training of students incl. theses etc. It draws attention to the necessity of organizational changes in the activities of the District hygiene station, to increased demands on the professional standard of workers and the need of close collaboration of field and laboratory departments. PMID- 10838781 TI - [History of the J.E. Purkyne Czech Medical Society]. AB - The author presents some basic data from the time of the foundation of the Czech Physicians Society (1862), the predecessor of the present Czech J. E. Purkyne Medical Association. Its beginnings date back to the year 1949. The author mentions also data associated with the foundation of the present Society of Epidemiology and Microbiology of the Czech J. E. Purkyne Medical Association. PMID- 10838782 TI - Synthetic aspects of iridoid chemistry. PMID- 10838783 TI - The defensive chemistry of ants. PMID- 10838784 TI - The Alpbach Public Health Workshop--a possibility for open interdisciplinary communication. PMID- 10838785 TI - [Suffering without meaning? Illness, pain and death. Development of evolutionary medicine]. AB - The human species is an optimized product of evolution. This realization and knowledge of the highly sensitive and sensible biocybernetical processes in our bodies is now largely out of sight in modern medicine. To develop an evolutionary and cross-cultural perspective of human life and its biological, psychic and social conditions is therefore particularly important for the health sciences. Birth behaviour, intensive care for premature babies, post-partum dysphoria, sudden infant death, hip dysplasia, cry babies, eating disorders, stress-induced diseases, depression, ageing and dying serve as examples to sketch this new approach. It may probably facilitate diagnostic and therapeutic schemes which will be better adapted to human needs and capacities and can, therefore, be more successful than the ones in the past. PMID- 10838786 TI - [Drug use: a life choice or health threat]. AB - The term "enjoyment" is used to denote a pleasurable, utility-creating activity or an act of consumption, which, however, may often be accompanied by subsequent risks, health-care costs or reduction in the quality of life. If viewed from the vantage point of health policy, we may quote several examples of consumer behaviour of this type, ranging first and foremost from the consumption of alcohol and nicotine to the use of "hard" drugs. But certain sports activities or sexual behaviour that may prove damaging to other persons must also be mentioned in this context. All these forms of consumption often lead to direct or indirect consequences to health and thereby result in increased health spending amounting to several thousand million DM. The decline in quality of life resulting from the above-mentioned forms of behaviour could be avoided through health promotion and prevention or, at the very least, through measures of rehabilitation, above all, information, counselling, and education, which represent medium- or long-term measures to that effect. Financial incentives or disincentives, for example, via an increase in co-payment in case of illness caused by risky behaviour, have not been as effective as had been expected by many proponents. Product-specific levies, risk-related payments and other financial incentives and disincentives are generally also not as effective as desired. Health policy measures aimed at influencing behaviour via financial measures are particularly ineffective in case of persons addicted to particular types of consumables, especially to drugs: Information, counselling and education represent non-financial forms of influencing and guiding behaviour with a greater scope for success. PMID- 10838787 TI - [European health systems and the integration problem of modern societies]. AB - With reference to the national health systems in Germany and the UK we must acknowledge that it was in particular Bismarck's Reform, originally directed toward a solidarity among the socially weak, which entailed in its development a marked redistribution via progressive health fees and standardized health services. In view of Alfred Marshall's original expectations this has resulted in a specific integration of the socially weak and with some difference for nationally tax-financed and social security financed health systems to a genuine contribution towards integration of modern society. An open research question is whether as a consequence of solidarity and integration through health systems there is a decline of social inequality for health. Equally open is the question as to the socio-structural and economic consequences the expansion of modern health systems has. PMID- 10838788 TI - [Multi-morbidity--fate or chance?]. AB - The increase of life expectancy in post-industrial societies also entails an increase in multimorbidity. Self-determination of the population and prevention through health organisations are imperative. In this connection different health concepts are discussed, although in most cases theoretical concepts concerning the health and illness behaviour of the population are missing. Present data show that the sphere of social epidemiology must be re-orientated. To this end, the behavioural science must also be taken into consideration in an appropriate manner. Many of the present, often activistic health-programmes are ineffective with regard to long-term impacts. There is a need to sound a warning in respect of an exaggerated economization of the health system: Self-responsibility and self-participation of patients alone cannot substantially stop the largely self generating costs, which result from medical-technological progress on the one hand and from pretentious demands on the other hand. PMID- 10838789 TI - [1000 illnesses--one health?]. PMID- 10838790 TI - [Personal responsibility--prevention]. PMID- 10838791 TI - [Personal responsibility--prevention]. AB - The multifarious structural web of modern health systems is endangered by political pressure, which leads to changing priorities and this renders it difficult, if not impossible to formulate an appropriate and long-term health policy. Particular risk factors are isolated from the complex connections and used as guidelines for prevention and sanction-measures. Considering the rising multimorbidity we should rather speak of risk indicators. There is an urgent need for scientific observation and foundation of preventive measures to minimise, or if possible stop, misinterpretations of data from short-term health research. Comprehensive health research is non-existent, responsibilities being unsettled and the necessity of long-term thinking has not been realised so far. Therefore, epidemiological studies considering pathogenetic and salutogenetic aspects to the same extent, are mandatory. PMID- 10838792 TI - [Personal responsibility--prevention]. PMID- 10838794 TI - [1,000 illnesses--one health? Multi-morbidity as responsibility]. PMID- 10838793 TI - [Personal responsibility and prevention in the tension field of health care systems]. AB - Thesis 1. The health care system determines the framework in each case. Thesis 2. Traditional health care systems impede self-responsibility and prevention. Thesis 3. New system elements are essential and possible. Thesis 4. In a given overall frame new systems have to set up with the responsibility of the individual promulgator. Thesis 5. These are the "clever" systems in the sense of European Managed Care. PMID- 10838795 TI - [The physician's image 2000]. PMID- 10838796 TI - [The physician's image 2000--requirements of education]. AB - The duties of the "Physician 2000 A.D." will continue to be determined by the needs of curative medicine. Therefore, in spite of many changes in medicine and in society as a whole, care for the individual patient will remain the predominant obligation of the future medical doctor. In order to comply with these obligations, the doctor must command a great power of judgement. To develop this faculty, apart from the acquisition of factual knowledge, empathy and a careful training in the art of critical observation are required. The present organisation of medical education in Germany over-states the acquisition of factual knowledge to some extent and under-rates the importance of empathy and the need for systematic training of the faculty of critical observation. This faculty can only be fully developed if it is trained right from the beginning of a medical student's education. In Germany changes in the regulations governing medical education are long overdue. This is particularly so with respect to the present system of examinations which over-emphasizes the results obtained by answering multiple-choice questions. PMID- 10838797 TI - [The physician's image 2000]. PMID- 10838798 TI - [The substance of medical education, graduate and continuing education, especially regarding specialty boards--a contribution to the physician's image of tomorrow]. PMID- 10838799 TI - [Multi-morbidity as a responsibility: 1,000 illnesses--one health. The physician's image 200]. AB - The prototype physician 2000 A.D. will have to be able to recognise multimorbidity and its individual importance in a general medical and social sense. There will be a specifically challenge in the fields of individual prophylaxis, interdisciplinary communication, integration of all nursing and treating partners and the ethical judgement between individually useful and theoretically possible treatment options. Medical education and social systems will have to provide enough knowledge and skills as well as opportunities to allow this necessary development of our future physicians 2000 A.D. PMID- 10838800 TI - [Managing quality problems in public health: role of the consumers]. PMID- 10838801 TI - [Health sciences--health occupations: lay system--professional system]. PMID- 10838802 TI - [Significance of quality assurance as an instrument for improving integrating and coordinating health care]. PMID- 10838803 TI - [Health sciences--health occupations: lay systems--professional systems]. PMID- 10838804 TI - [In Process Citation] PMID- 10838805 TI - Improvement of accuracy of chromosome aberration analysis for biological radiation dosimetry. AB - The frequency of chromosome aberrations in circulating lymphocytes is accepted as being the most reliable indicator of the absorbed dose of radiation. Researches done to improve the accuracy of cytogenetic analysis are described in this review. These include investigations of in vitro factors that affect the yield of radiation-induced aberrations and of in vivo factors that affect the chromosomal radiosensitivity of individuals. Improved chromosome-painting methods for accurate judgment of dicentrics and translocations are introduced. The practicality of these advanced cytogenetic techniques is shown by examinations of individuals exposed in the radiation accident at Tokaimura in 1999. PMID- 10838806 TI - Positional cloning and functional analysis of the gene responsible for Nijmegen breakage syndrome, NBS1. AB - Nijmegen breakage syndrome (NBS) is a rare autosomal recessive disorder characterized by microcephaly, combined immunodeficiency, and a high incidence of lymphoid tumor. Cells from NBS patients show chromosomal instability, hypersensitivity to ionizing radiation and abnormal p53-mediated cell cycle regulation. We cloned the underlying gene for NBS, designated NBS1, by complementation-assisted positional cloning from the candidate region 8q21. Large genomic sequencing, as well as a search using computer programs, provides a powerful approach for identifying the underlying gene for a disease. The NBS1 gene encodes a protein of 754 amino acids that has FHA and BRCT domains which often are conserved in cell-cycle checkpoint proteins. The gene has weak homology to the yeast (Saccharomyces cerevisiae) Xrs2 protein in the N-terminus region. Like yeast Xrs2, the NBS1 protein forms a complex with hRAD50/hMRE11, and the complex is condensed as foci in the nucleus after irradiation, indicative that this triple-complex is a crucial factor in DNA repair. Functional analysis of the NBS1 protein is in progress and it should provide further clues to understanding the repair mechanism of radiation-induced DNA double-strand breaks. PMID- 10838807 TI - Purification and characterization of a novel DNA repair enzyme from the extremely radioresistant bacterium Rubrobacter radiotolerans. AB - Rubrobacter radiotolerans is an extremely radioresistant bacterium. It exhibits higher resistance than the well-known radioresistant bacterium Deinococcus radiodurans, but the molecular mechanisms responsible for the radio-resistance of R. radiotolerans remain unknown. In the present study, we have demonstrated the presence of a novel DNA repair enzyme in R. radiotolerans cells that recognizes radiation-induced DNA damages such as thymine glycol, urea residues, and abasic sites. The enzyme was purified from the crude cell extract by a series of chromatography to an apparent physical homogeneity. The purified enzyme showed a single band with a molecular mass of approximately 40 kDa in SDS-polyacrylamide gel electrophoresis, and was designated as R-endonuclease. R-Endonuclease exhibited repair activity for thymine glycol, urea residues, and abasic sites present in plasmid DNA, but did not act on intact DNA, UV-irradiated DNA and DNA containing reduced abasic sites. The substrate specificity together with the salt and pH optima suggests that R-endonuclease is a functional homolog of endonuclease III of Escherichia coli. PMID- 10838808 TI - Thyroid abnormality trend over time in northeastern regions of Kazakstan, adjacent to the Semipalatinsk nuclear test site: a case review of pathological findings for 7271 patients. AB - From 1949 through 1989 nuclear weapons testing carried out by the former Soviet Union at the Semipalatinsk Nuclear Test Site (SNTS) resulted in local fallout affecting the residents of Semipalatinsk, Ust-Kamenogorsk and Pavlodar regions of Kazakstan. To investigate the possible relationship between radiation exposure and thyroid gland abnormalities, we conducted a case review of pathological findings of 7271 urban and rural patients who underwent surgery from 1966-96. Of the 7271 patients, 761 (10.5%) were men, and 6510 (89.5%) were women. The age of the patients varied from 15 to 90 years. Overall, a diagnosis of adenomatous goiter (most frequently multinodular) was found in 1683 patients (63.4%) of Semipalatinsk region, in 2032 patients (68.6%) of Ust-Kamenogorsk region and in 1142 patients (69.0%) of Pavlodar region. In the period 1982-96, as compared before, there was a noticeable increase in the number of cases of Hashimoto's thyroiditis and thyroid cancer. Among histological forms of thyroid cancer, papillary (48.1%) and follicular (33.1%) predominated in the Semipalatinsk region. In later periods (1987-96), an increased frequency of abnormal cases occurred among patients less than 40 years of age, with the highest proportion among patients below 20 in Semipalatinsk and Ust-Kamenogorsk regions of Kazakstan. Given the positive findings of a significant cancer-period interaction, and a significant trend for the proportion of cancer to increase over time, we recommend more detailed and etiologic studies of thyroid disease among populations exposed to radiation fallout from the SNTS in comparison to non exposed population. PMID- 10838809 TI - High incidence of micronuclei in lymphocytes from residents of the area near the Semipalatinsk nuclear explosion test site. AB - The Semipalatinsk area is highly contaminated with radioactive fallout from 40 years of continuous nuclear testing. The biological effects on human health in this area have not been studied. Significant remaining radioactivities include long-lived radioisotopes of 238,239,400Pu, 137Cs and 90Sr. To evaluate the long term biological effects of the radioactive fallout, the incidence of micronuclei in lymphocytes from residents of the area was observed. Blood was obtained from 10 residents (5 females and 5 males, aged 47 to 55 years old) from each of the 3 areas of Znamenka, Dolon and Semipalatinsk, which are about 50-150 km from the nuclear explosion test site. For micronucleus assay, PHA-stimulated lymphocytes were cultured for 72 h and cytochalasin B was added at 44 h for detecting binuclear lymphocytes. Five thousand binuclear lymphocytes in each resident were scored. The means of micronucleus counts in 1,000 lymphocytes in residents of Semipalatinsk, Dolon and Znamenka were 16.3, 12.6, and 7.80, respectively, which were higher than those of the normal Japanese persons (4.66). These values were equivalent to the results obtained from 0.187-0.47 Gy of chronic exposure to gamma-rays at a dose rate of 0.02 cGy/min. The high incidence of micronuclei in residents of the Semipalatinsk nuclear test site area was mainly caused by internal exposure rather than external exposure received for the past 40 years. PMID- 10838810 TI - The role of the sympathetic nervous system in radiation-induced apoptosis in jejunal crypt cells of spontaneously hypertensive rats. AB - To evaluate the effect of the sympathetic nervous system on radiation-induced apoptosis in jejunal crypt cells, apoptosis levels were compared in spontaneously hypertensive rats (SHR), animals which are a genetic hyperfunction model of the sympathetic nervous system, and normotensive Wistar-Kyoto rats (WKY). SHR and WKY were exposed to whole body X-ray irradiation at doses from 0.5 to 2 Gy. The apoptotic index in jejunal crypt cells was significantly greater in SHR than in WKY at each time point after irradiation and at each dose. WKY and SHR were treated with reserpine to induce sympathetic dysfunction, and were subsequently exposed to irradiation. Reserpine administration to SHR or WKY resulted in a significant suppression of apoptosis. p53 accumulation was detected in the jejunum in both WKY and SHR after irradiation by Western blotting analysis. There were no significant differences in the levels of p53 accumulation in irradiated intestine between WKY and SHR. These findings suggested that hyperfunction of the sympathetic nervous system is involved in the mechanism of high susceptibility to radiation-induced apoptosis of the jejunal crypt cells. PMID- 10838811 TI - [Changes in some of the kallikrein-kinin system indices in patients with acute pancreatitis]. AB - Proceeding from the major role played by kinins in the pathophysiology of endogenic intoxication among acute pancreatitis patients (AP), and the conflicting and scarce literature data on the issue, the changes in the level of prekallikrein, high-molecular kininogen, alpha 2-macroglobulin, alpha 1 antitrypsin, plasminogen and carboxypeptidase N in the blood are studied in dynamics at 1, 3, 5, 7, 9 and 14 days after admission of 48 patients with mild, and 121 with severe form of acute pancreatitis. Forty-eight individuals are used for control purpose. PK, KG and plasminogen are assayed using the colorimetric method of the Boehringer Company--Mannheim, KG--by chronometric test of the Sigma Diagnostics Company, CPO N--after Folk's method, as modified by Erdos, alpha 2-MG -by radial immunodiffusion according to Mancini, and alpha 1-AJ--by immunoturbidometric method. As shown by the results, in acute pancreatitis KKS activation occurs, demonstrated by the reduced PK, KG and alpha 2-MG values, and by the statistically significant enhancement of alpha 1-DJ, COP N and plasminogen activity. In patients presenting mild forms the aforementioned changes are rather weakly manifested and transient, while in the serious forms they are markedly expressed and persisting. In either form the deviations are rather pronounced in the first three days of disease. Coinciding with a clinical course characterized by cardiovascular changes similarly strongly manifested. PMID- 10838812 TI - [The early results of autogenous plastic reconstruction of the breast using the TRAM and LDM methods in patients with breast carcinoma. Transverse rectus abdominis muscle. Latissimus dorsi muscle]. AB - It is the purpose of the study to assess the early results of TRAM (transversus rectus abdominis musculocutaneous flap) and LDM (latissimus dorsi musculocutaneous flap) techniques used in mammary gland reconstruction following mastectomy in breast carcinoma patients. Over a two-year period (Nov 1995 through Nov 1997), nineteen patients presenting breast carcinoma undergo treatment. In ten immediate reconstruction of the breast is undertaken, and in nine the operation is deferred for varying periods of time. TRAM technique is used in 18 cases, and LDM--in one. The mean postoperative follow-up period is 12 months (range 1 to 24 months). Evaluation of results is done with a reference to local relapses and distant metastases, cosmetic outcome of the reconstruction and complications. There is conclusive evidence that in a considerable number of patients autogenous mammary gland reconstruction is practicable as totally complying with the oncologic tenet concerning life style improvement in this high risk contingent of patients. PMID- 10838813 TI - [The surgical treatment of pulmonary and associated echinococcosis in childhood]. AB - Over a nine-year period (1990-1998), a total of 196 children with pulmonary and associated hydatid disease, aged 2 to 16 years, undergo treatment in the Department of Pediatric Thoracic Surgery--Emergency Medicine Institute "N. I. Pirogov"--Sofia. One hundred thirty children (62.1%) present isolated, and sixty six (38.8%)--associated hydatid disease, of which with localization in the liver (63 cases), brain (1), kidney (1) and spleen (1). The pulmonary cysts are located in the left lung in 96 cases (48.9%), right lung--79 (40.3%) and bilateral involvement--21 (10.8%). In 74 children the disease runs an asymptomatic course, in 128 (65.3%)--with fever and cough, and in 65 (33.1%)--with thoracic pain. On admission 183 children (94.4%) are in a satisfactory and good general condition; 13 with complications in the cyst and pleural cavity are in a serious general condition accompanied by respiratory failure. In 129 children diagnosis is made on the ground of x-ray study, in 40 CAT study is additionally performed, and in 24--ultrasonography. Of 22 children with bilateral pulmonary location, 13 (59.9%) undergo operation in two stages, and nine (40.1%)--in one stage, with bilateral thoracotomy done in seven, and sternotomy in two instances. Intraoperatively more than one echinococcus cysts are discovered in 23 cases (11.7%). In 105 children (53.5%) echinococcotomy is performed, in 67 (34.2%)--atypical resection, in 8 (4.1%)--lobectomy, in 15 (7.6%)--segmentectomy and in one--pneumonectomy. In the early postoperative period, one child develops hemothorax from a bleeding intercostal artery, and another one--pneumothorax from unsutured bronchus in the cyst bed necessitating emergency re-thoracotomy. The outcome is fatal in one patient with malignant hyperthermia, and the remainder are discharged clinically cured. PMID- 10838814 TI - [The pathomorphology of complicated colorectal cancer and the stages in the development of the tumor process]. AB - A comprehensive pathomorphological assessment is done in 385 patients presenting complicated colorectal carcinoma, covering the period 1990 through 1995. The patients are distributed in 4 groups according to stage, with those in stage III being the most numerous--141 cases. The growth pattern of the neoplasm is as follows: exophytic growth--110 cases, ulcerous growth--57, cup-like growth--13, and endophytic growth--205 cases. On establishing metastatic lymph nodes within I and II lymph collectors, prophylactic postoperative chemo- and immunotherapy are undertaken. In case of lymph nodes present in III lymph collector, the operation is taken to be relatively radical, and for that reason chemo- and immunotherapy assume a therapeutic character. PMID- 10838815 TI - [The video-assisted thoracoscopic treatment of spontaneous pneumothorax--our experience with 42 patients]. AB - We present our experience with video-assisted thoracoscopic surgical (VATS) treatment of spontaneous pneumothorax (SP) in 42 consecutive patients. We performed 45 video-thoracoscopic procedures during 44 hospitalisations. The most common procedures were: Endo-stapling of bullae in 12 pts, 4 ligations of bullae, 8 suturing of bullae, 27 bleb ablations and catheterizations. In all patients a parietal pleural abrasion was performed. We describe a new variation of mechanical pleurodesis for prevention of recurrences--"linear pleurotomy"- performed in 15 patients. No major complication occurred during surgery. The mean operative time was 67 minutes (range 30 to 160 min). We have 2 conversions to thoracotomy in patients with secondary SP. Two early recurrences were treated one by thoracotomy and the other by re-thoracoscopy. One case of postoperative bleeding required re-thoracoscopy. Four patients had a prolonged air-leak (more than 7 days), but recovered without additional treatment. The mean duration of intercostal drainage was 4.64 days, the mean postoperative hospital stay--6.8 days. We report 95.6% success rate after VATS treatment of SP with mean follow-up period of 21.1 months (2-49 months). PMID- 10838816 TI - [Francke's operation--a resolution for deep small echinococcal cysts of the liver]. AB - The operation of Francke in hydatid disease treatment is described--puncture, aspiration, sterilization--with emphasis laid on the technical details. The indications for its practical implementation are specified--small echinococcus cysts, deeply located within the parenchyma of the organ, or diffuse abdominal echinococcosis. In diagnosing such cysts intraoperative ultrasonography is considered as absolutely necessary and mandatory. The operation is practicable in handling first-stage cysts according to Volynkin, containing thin echinococcus liquid only, whereas in the presence of daughter vesicles it is not indicated. Postoperatively, dispensary outpatient observation and chemotherapy of the hydatid disease are recommended with a view to promote radical cure. PMID- 10838817 TI - [The long-term results in complicated colorectal cancer (survival for over 3 and 5 years)]. AB - Colorectal cancer survival depends on: stage of primary neoplastic process development, clinical complication pattern and clinical course of the disease, type (radicalism) of the surgical intervention done and histological verification of blastoma. As shown by the results, postoperative lethality in complicated colorectal carcinoma cases amounts to 17.1 per cent, whereas in those undergoing radical surgery for uncomplicated carcinoma it amounts to 6.8 per cent. Patients presenting complicated colorectal carcinoma are admitted to the clinic with diagnosis advanced stage of development of the disease which explains the lower survival rate. The studies performed show that five-year survivorship in patients with complicated colorectal carcinoma amounts to 33.3 per cent, while among those operated for uncomplicated colorectal carcinoma it is 62.7 per cent in the average. PMID- 10838818 TI - [The topographic anatomical aspects of some kidney anomalies]. AB - On 50 cases of the dissecting material of the Department of Anatomy and Histology were worked out the kidney's vessels, the basin and the ureters, as the varieties in number and situation of the vessels, the cases of nephric ectopy and comparatively rare case of combined anomaly as well were traced and described. This anomaly is expressed in bilateral nephric ectopy, bilateral double ureter, which begins from independent calices set, and four pairs of polar kidney arteriae, which are situated bilateral as well. In the right they pass before v. cava inf. The vessels are equally calibrated, as the three pairs are branches of aorta abdominalis and the fourth of a. iliaca comm. The cases which were described attract the surgeons' interest because the zone of the kidney, its vessels and the way of the ureter are an object of frequent surgical interventions. PMID- 10838819 TI - [The diagnostic and surgical problems in ureter fissum]. PMID- 10838820 TI - [The pathomorphological changes of the retroperitoneum in acute pancreatitis]. AB - Attention is focused on the development of a news idea. A comparative assessment of the body tissue tegmina--peritoneum and skin--is performed using a precise measuring unit--the square centimeter. Rating of pathological changes in the retroperitoneum in acute pancreatitis, similar to the one in thermal or chemical burns of skin, expressed in percents corresponding to the square centimeters of pathologically altered surface, is proposed. PMID- 10838822 TI - [A case of combined echinococcosis with a clinically unclear impaired renal drainage function]. PMID- 10838821 TI - [Intrascrotal nontesticular tumors]. AB - This is a review of literature data on rarely met with intrascrotal non testicular tumors still not well enough clarified in clinical practice. Histologically these tumors derive from: 1. Epididymis. 2. Funiculus spermaticus and/or tunica vaginalis. 3. Tissues within the scrotum (fatty, fibrous, muscular, lymphatic, nervous). 4. Metastases. The individual types of neoplasms, some of them representing casuistic rarity, so far described as sporadic observations in the pertinent literature, are discussed. Adenomatoid tumor of epididymis and rhabdomyosarcoma are the most frequently encountered. Clinically paratesticular tumors do not lend themselves to differentiation from the ones involving the testis regardless of the fact that for some of them data on specific laboratory and serum or tissue tumor markers have been duely reported. Diagnosis is made histologically following operative management--orchiepididymectomy. According to histological pattern, if necessary the treatment proceeds with radio- or chemotherapy. Finally, two personal observations are described. PMID- 10838823 TI - [Rather rare cases of echinococcal cyst location]. PMID- 10838824 TI - [Lipoma of the temporal lobe]. AB - This is a case report on a 16-year-old girl presenting lipoma located in the temporal cerebral lobe, in conjunction with an ever increasing frequency of epileptic seizures, unresponsive to the anticonvulsive treatment carried out. A comprehensive analysis is done of the data concerning clinical course, diagnostic examinations' specificity and microsurgical removal of the lipoma with ensuing optimal postoperative results--total abatement of epileptic seizures against the background of reduced anticonvulsive therapy throughout the 3.5 years observation period. The literature survey shows that intracranial lipomas with the location described represent as casuistic rarity. PMID- 10838825 TI - [Acute surgical abdomen in gallbladder cancer]. PMID- 10838826 TI - [A rare combination of hepatic echinococcosis, cholelithiasis and gangrenous cholecystitis]. PMID- 10838827 TI - [Clinico-morphological problems in peliosis hepatis]. PMID- 10838828 TI - [Complications in gallstones]. PMID- 10838829 TI - [Chronic heart failure. Diagnosis and therapy]. PMID- 10838830 TI - [Interventional treatment of coronary heart diseases. Feasibility of percutaneous catheterization techniques]. PMID- 10838831 TI - [Burning mouth syndrome]. PMID- 10838832 TI - [Hepatitis B. Recognition, treatment and prevention]. PMID- 10838833 TI - Plasma levels of E-selectin in normolipemic and hyperlipemic arteriopathic patients after vasoactive and lipid-lowering treatment. AB - BACKGROUND: The authors studied the plasma levels of E-selectin in a group of arteriopathic patients, before and after vasoactive and lipid-lowering treatment. METHODS: The series consisted of 73 subjects (53 males, 20 females, aged 54 +/- 9 suffering from occlusive peripheral arteriopathy; 21 subjects with total cholesterol (TC) below 200 mg/dl were considered as normolipemics (group A); 24 subjects with TC between 200 and 240 mg/dl, mild hypercholesterolemics (group B); 18 with TC above 240 mg/dl, severe hypercholesterolemic (group C); 10 subjects who had high triglyceride values (above 200 mg/dl), (group D); 12 normal controls were also considered. All patients underwent a vasoactive treatment for 15 days; group B also underwent a standard hypolipidic diet (phase I NCEP, lipid 30% die) groups C and D underwent the same diet associated respectively with simvastatin (200 mg/die) and bezafibrate (400 mg/die). For each sample of plasma before and after treatment the determination of levels of E-selectin was carried out by an immunoenzymatic method (kit ELISA Amersham). RESULTS: In groups A-B-C-D a reduction of the plasma levels of E-selectin was found, which was significant (p < 0.05), for group C and D, compared to controls. In groups A-B-D significant changes of E-selectin were not found after treatment; in group C the difference between the values after treatment and at baseline was significant (p < 0.05). CONCLUSIONS: The reduction of the E-selectin plasma levels was proportional to the values of total cholesterol and triglycerides; the chronicity and the dyslipidemia may be responsible for the basal diminished biosynthetic endothelial function; in the severe hypercholesterolemics the lipid-lowering treatment caused a significant decrease of E-selectin, due to a probable reduced endothelial irritation dependent on the hyperlipemic stress. PMID- 10838834 TI - Pulse wave velocity. A new calculation method. AB - BACKGROUND: The increase in aortic Pulse Wave Velocity (PWV) is considered a surrogate marker of vascular disease; it can be non-invasively assessed by means of an indirect method calculating the time that the pulse wave takes to travel a definite distance along the vasculature; the distance/time ratio corresponds to the velocity measure. The presentation of a new calculation method is the aim of the present study. METHODS: A duplex scanner at the common carotid artery, the abdominal aorta at the prebifurcation site and the femoral common artery levels, was performed on 127 out-patients with risk factors, 38 of which were affected by clinical vascular disease, and on 50 healthy subjects (control group). The spectral analysis from these three sites was registered simultaneously with an ECG trace and the interval between the R wave apex and the spectral complex systolic foot was measured. The Transit Time (TT) was calculated by the difference between the values obtained from the proximal and distal measurement sites and PWV dividing the distance between them by the TT (PWV = Distance/TT); statistical significance and intra and inter observer variation coefficient, expressed as mean +/- standard deviation, were calculated by the analysis of variance and Turkey test, the correlation with the major risk factors and the intima-media thickness by the multivariate analysis. RESULTS: The PWV is increased in the patients group compared to control group (p < 0.001). Multivariate analysis shows a positive correlation with age, hypercholesterolemia, arterial hypertension, diabetes mellitus, intima-media thickness and arterial disease, no correlation was found with the smoking habit. The "patients" group shows an increased PWV in those with atherosclerotic plaques and/or clinical vascular disease compared to non atherosclerotic patients with risk factors (p < 0.001); the intra and inter observer variation coefficient ranged between 4.87 +/- 1.82% and 8.06 +/- 3.16% respectively. CONCLUSIONS: The proposed PWV measurement method is simple, quick, reproducible and repeatable, it can separate healthy subjects from patients with risk factors, atherosclerotic plaques and clinical vascular disease. Due to the strong correlation with age, normal values in different age groups are necessary before the PWV can be used as a marker of vascular disease. PMID- 10838835 TI - Color-Doppler sonography in ophthalmology. AB - BACKGROUND: With the present study we considered ocular blood flow by the use of color Doppler ultrasound. We compared the most clinically important arteries of both eyes. We valued the central retinal artery, the posterior long ciliary arteries, the posterior brief ciliary arteries and the central retinal vein. METHODS: We submitted to an ecd examination of the ophthalmic arteries 50 patients, of whom 30 were healthy, 4 presented a pallor of the optic disk, 6 were affected by chronic simple glaucoma and 10 were affected by arterial hypertension. RESULTS: This study has allowed us to define the normal and pathologic anatomic findings at the ecd tracing therefore qualitative and quantitative standards of this examination, repeatable and comparable. CONCLUSIONS: Currently the different techniques used to study ocular blood flow, do not allow its exact detection, but only a morphological evaluation of the eye artero-venous system. Acquisition of morphological and velocimetric data, at the same time, allows a more exact evaluation of the pathology. Therefore the ecd is a simple rapid and non-invasive approach, useful in the acquisition of important data concerning diagnostic confirmation, prognosis, and above all pharmacological follow-up of many pathologies such as diabetes, glaucoma and arterial hypertension. The diagnosed vascular alterations are correlated to the degree retinopathy and to the effectiveness of the pharmacological treatment. PMID- 10838836 TI - Long-term follow-up after syncope. A group of 183 patients observed for 5 years. AB - BACKGROUND: To determine the incidence of recurrent syncope and mortality rate in a group of patients hospitalized for syncope. METHODS: A 5 years follow-up of 183 patients hospitalized for syncope. A collaborative study between the Departments of Cardiology and Neurology. RESULTS: The etiological diagnosis of syncope was the following: unknown causes 21.86%, cardiovascular causes 72.67%, non cardiovascular cause 5.46%. The general mortality rate was 26.77% (51.94% in those aged > = 70 years, 8.49% in < 70 years). The mortality rate of syncope of unknown causes was 30% among all patients (61.11% in those aged > = 70 years and 4.54% < 70 years). Syncope of cardiac cause (prevalent arrhythmias) and syncope of iatrogenic cause had a high mortality rate (respectively 63.33% and 42.10%); both in young people (28.53%, 26.53%) and in old people (66.66%, 50%). Syncope of reflex cause has a mortality rate of 5.79% (4/69) among all patients (14.28% in those aged > = 70 years, 3.63 in < 70 years). Age, arterial hypertension, ischemic heart disease and cerebrovascular events are significantly associated with mortality rate. Recurrences were quite common: 24.59% of all patients (45/183); 29/45 syncope (64.44%) were observed in the first year of follow-up with a prevalence for patients with a syncope of reflex cause (26.09%) and for those with a syncope of unknown cause (37.56%). Recurrences were less common among patients with syncope of cardiac cause, however more frequent during the first year of follow-up, with an increased risk of mortality. CONCLUSIONS: A. The cause of syncope is most frequently established on the basis of history and clinical examination. B. A collaboration between Departments with a common interest for this pathology is recommended. C. Cardiac syncope has the worst prognosis and therefore needs recurrent clinical examinations and prompt treatment. D. Syncope itself is not a risk factor for increased overall and cardiac mortality or cardiovascular events. E. Underlying diseases such as hypertension, ischemic heart diseases, congestive heart failure, cerebrovascular events are the major risk factors for mortality. PMID- 10838837 TI - Cardiac damage following therapeutic chest irradiation. Importance, evaluation and treatment. AB - Radiation induced heart disease, with its clinical manifestations, is becoming a growing problem. Its prevalence is increasing, keeping pace with the increased survival of many malignancies. The majority of patients with radiation induced heart disease is constituted by Hodgkin's disease survivors, followed by non Hodgkin's disease, esophageal carcinoma, thymoma, lung cancer, breast cancer and metastatic seminoma. Pericardial disease is the most well known expression of radiation induced heart disease, although the whole cardiac structure is compromised because of the structural and consequently functional impairment. Myocardial damage can lead to a congestive heart failure, typically due to a restrictive cardiomyopathy. Coronary artery obstructive disease frequently involves ostial coronary segments and the left main, for this reason it does appear particularly harmful. All patients undergoing chest irradiation require serial cardiological evaluation. Important risk factors of radiation induced heart disease are previous chemotherapy, radiation exposition exceeding 4000 Rad, administration next to the heart and on the left side of the chest must be taken into particular consideration. The cardiac damage limitation basically is founded on prevention. Significant results have been obtained with fractional exposition, high energy utilization and "split" zone covering. The radiotherapeutic technical improvement with the comprehensive individual patient risk evaluation will provide a substantial benefit for the future. The consultant cardiologist should cooperate with the oncologist and the radiotherapist, providing specific competence and continuative care. PMID- 10838838 TI - Aortic dissection after superior mesenteric artery percutaneous stenting. Case report. AB - We report an unusual case of aortic dissection after superior mesenteric artery percutaneous stenting. A 44-year-old patient, who suffered from back pain and fever, was diagnosed as having an aortic dissection. Aortic dissection, extending from the aortic arch (just after left subclavian artery origin) to the aortic carrefour, was successfully diagnosed by means of Duplex scan and CT scan examination. Two pathogenetic hypotheses, malformative and iatrogenic, were discussed. PMID- 10838839 TI - Green and brown colour effects in tremellaceous yeast fungi on Staib agar. AB - Fifty-eight species of basidiomycetous yeast fungi were examined for green and brown colour effects (GCE, BCE) on Staib agar (Guizotia abyssinica creatinine agar). In addition to 9% of Cryptococcus laurentii strains tested, only Cryptococcus podzolicus was GCE-positive. Out of 14 strains of this species, 13 showed a GCE and four showed a BCE. All GCE-positive strains utilized creatinine and creatine as a source of nitrogen but not of carbon. None of these strains could grow at 37 degrees C. Unlike Cr. laurentii, no strains of Cr. podzolicus assimilated glycine as a source of carbon. No association was found to exist between GCE and killer activity. Some tremellaceous creatinine-negative species (Cryptococcus gastricus) showed a BCE whereas some sporidiobolaceous creatinine positive yeasts (Leucosporidium, Rhodotorula) produced neither GCE nor BCE. PMID- 10838840 TI - Occurrence of Cryptococcus spp. in excreta of pigeons and pet birds. AB - In pooled samples of faeces from 25 pet bird flocks in Thuringia, a high rate of contamination with Cryptococcus neoformans var. neoformans was found. The prevalence of Cr. neoformans in the bird-breeding establishments correlated with the numbers of the different pet bird species in these flocks. The differentiation between varieties of Cr. neoformans by means of proline assimilation and canavanine resistance detection as well as with the aid of Cr. neoformans factor sera, polymerase chain reaction (PCR) fingerprinting, sequencing of PCR products as well as Fourier transform infrared spectroscopy showed uniform results which also corresponded to the serological differentiation between serovars A and D. A predominance of serovar A could be observed among the pet bird breeding flocks. This corresponded to the frequency distribution of serovars A and D in cases of human diseases in Germany. In 50% of the samples of pigeon excreta examined (n = 30) in Innsbruck (Austria), Cryptococcus albidus could be isolated but not Cr. neoformans. However, this Cryptococcus species is of minor pathogenetic importance for man. Cryptococcus albidus may be clearly distinguished from Cr. neoformans by means of microbiological methods, PCR and Fourier transform infrared spectroscopy. PMID- 10838841 TI - Inhibitory effect of essential oils on apical growth of Aspergillus fumigatus by vapour contact. AB - The inhibitory effect of seven essential oils on the apical growth of hyphae of Aspergillus fumigatus was studied using a bio cell tracer by vapour contact in a sealed vessel. Based on the inhibitory pattern, these essential oils were classified into three groups. The first group, composed of citron, lavender and tea tree oils, stopped the apical growth in a loading dose of 63 micrograms ml-1 air, but allowed the regrowth of the hyphae after removal of the vapour, indicating fungistatic action. The second group, consisting of perilla and lemon grass oils, stopped the apical growth in a loading dose of 6.3 micrograms ml-1 air, and did not allow the regrowth after gaseous contact at 63 micrograms ml-1 air, indicative of fungicidal action. The third group, consisting of cinnamon bark and thyme oils, retarded the growth in a dose of 6.3 micrograms ml-1 air, stopped it in a dose of 63 micrograms ml-1 air, and incompletely suppressed regrowth of the hyphae. Gas chromatographic analysis revealed that vapours of essential oils were absorbed on fungal mycelia and agar medium most abundantly by the first group, followed by the second and third groups, reflecting the volatility of the respective groups. Suppression of the apical growth by vapour contact was ascribed to the direct deposition of essential oils on fungal mycelia, together with an indirect effect via the agar medium absorbed. PMID- 10838842 TI - Evaluation of the growth dynamics of Trichophyton rubrum cultures by morphometry and non-linear curve fitting analysis. AB - To determine the size of dermatophyte thalli, a point-counting procedure with a square lattice grid is proposed instead of using the one-dimensional diameter as the size parameter. For an analysis of the growth dynamics of such colonies, the area values might be subjected to a non-linear curve-fitting procedure. To demonstrate the method, samples of 15 Trichophyton rubrum strains, isolated from human skin and cultured concomitantly on Sabouraud and Selective agar during a period of 8 weeks, were used. The morphometric assessment of the area of the thalli, subjected to non-linear curve fitting, disclosed only a slight difference in the growth dynamics. PMID- 10838843 TI - Non-dermatophytic onychomycosis. An understimated entity? A study of 51 cases. AB - The aim of our study was to evaluate the incidence, the clinic characteristics, the therapeutic antifungal response and the evaluation of side-effects in 51 non dermatophytic onychomycosis cases which were observed in a case-study of 1012 patients, affected by different types of onychopathy, attending the Mycology Ambulatory of San Paolo Hospital, Milan, Italy during the period 1994-97. PMID- 10838844 TI - Intermittent pulse therapy with itraconazole for onychomycosis. Experience in Hong Kong Chinese. AB - Forty-four Hong Kong Chinese adults comprising 10 cases of fingernail and 42 cases of toenail onychomycosis were recruited for a 3-weekly itraconazole pulse therapy. Each pulse consisted of seven consecutive daily 400 mg doses with a 3 week interval between treatments. All patients in the fingernail group and 37 in the toenail group completed the study. The clinical cure, clinical response and mycological cure rates at week 36 were 70, 90 and 90% for fingernail and 35, 81 and 68% for toenail groups, respectively. Side-effects including biochemical abnormalities were minimal, and returned to normal upon cessation of treatment. We conclude that itraconazole pulse therapy is very effective, safe and well tolerated for fingernail and toenail onychomycosis. PMID- 10838845 TI - The frequency of tinea pedis in patients with tinea cruris in Tehran, Iran. AB - The objective of this study is to define the frequency of tinea pedis in patients with tinea cruris. Sixty patients (46 males and 14 females) entered our study. Epidermophyton floccosum was the most frequently isolated fungus in tinea cruris. Four patients (6.4%) had concurrent tinea pedis and the only isolate from the foot was Trichophyton mentagrophytes. In three of them, the corresponding dermatophyte in the groin was E. floccosum. PMID- 10838846 TI - Adhesion of dermatophytes to healthy feet and its simple treatment. AB - At several public baths, we isolated dermatophytes from the soles of healthy volunteers by a new direct isolation method (foot-press culture method). We confirmed that a public bath is one of major sources of infection of dermatophytes. We showed that simple treatments such as (i) wiping the sole with a towel; (ii) washing with soap; (iii) 100 steps on another mat; and (iv) holding the foot up for an hour, significantly reduced the fungi on the soles of six healthy volunteers. These treatments may be effective for prevention of tinea pedis. PMID- 10838847 TI - Mycological examination of the hair samples of 11 school-going Nepalese children suspected of tinea capitis. AB - Among 150 students from a semi-urban school in Kathmandu (Nepal), 30 from various age groups and both sexes with symptoms suggestive of scalp ringworm, were examined clinically. Hair samples were collected for mycological examination, using the hair brush method, from 11 of the 30 children (36.7%), aged between 5 and 14 years, who were clinically diagnosed. Four of the cultured samples were positive for isolates of Trichophyton violaceum. This minor study provides evidence that tinea capitis might constitute a substantial infectious dermatological problem in Nepal. The study also indicated T. violaceum as the causative organism of tinea capitis. PMID- 10838848 TI - Isolation of dermatophytes and other keratinophilic fungi from soils of Mysore India. AB - In Mysore, India, 88 soil samples were collected from various areas and habitats and screened for the presence of dermatophytes and related keratinophilic fungi. PMID- 10838849 TI - Case report. Mixed systemic mycosis with fatal outcome in a patient with acute myeloblastic leukaemia. AB - Fungal infections represent an increasing problem in immunocompromised patients. The majority of cases are caused by one single fungal pathogen and infections with more than one fungus are very rare. Here we describe a case of combined infection with Aspergillus and a zygomycete species, involving the lungs, spleen and the brain and leading to fatal outcome in spite of early antimycotic therapy. PMID- 10838850 TI - Case report. Fatal Aspergillus flavus pericarditis in a patient with acute myeloblastic leukaemia. AB - We report a case of Aspergillus flavus pericarditis treated with fluconazole for oral candidosis. The patient with acute myeloblastic leukaemia developed tachypnoea after antileukaemic chemotherapy. Pericardial effusion was seen in the echocardiogram. Aspergillus flavus was isolated from the pericardial fluid. The patient died from aspergillosis, before the antimycotic treatment could be changed to amphotericin B. PMID- 10838851 TI - Case report. Meningitis due to Cryptococcus neoformans var. neoformans serotype AD associated with pulmonary tuberculosis. AB - A case of meningitis in a 38-year old male with pulmonary tuberculosis is described. Cryptococcus neoformans was isolated from the cerebrospinal fluid. The isolate belonged to variety neoformans serotype AD. The patient died before any antimycotic therapy could be instituted. PMID- 10838852 TI - Case report. Granuloma caused by Cryptococcus neoformans. AB - A case of cerebral cryptococcal granuloma is reported. The diagnosis was based on histological examination and rDNA-PCR identification. Complete cure was achieved with fluconazole. PMID- 10838853 TI - Case report. An unusual case of cutaneous sporotrichosis and its response to weekly fluconazole. AB - Cutaneous sporotrichosis is clinically divided into two main types: lymphocutaneous and fixed plaque type. Our report represents an unusual case with both types simultaneously. Fluconazole 150 mg once weekly was used as the treatment. The fixed type lesions responded very well and were healed after 4 months, but the lymphocutaneous lesions were not controlled even after 6 months of the treatment. PMID- 10838854 TI - Case report. A case of chromoblastomycosis effectively treated with terbinafine. Characteristics of chromoblastomycosis in the Kitasato region, Japan. AB - A 38-year-old male with history of trauma in the left gluteal region 20 years ago presented with a dark red skin eruption at the traumatized area. It gradually grew to form an erythematous plaque with a well-defined border. Clinical findings and mycological cultures resulted in the diagnosis of chromoblastomycosis due to Fonsecaea pedrosoi. After initial administration of 5-fluorocytosine and local heat an almost complete cure was achieved with terbinafine combined with local heat therapy. A review is given on the chromoblastomycosis cases observed in the Kitasato region in Japan. PMID- 10838855 TI - Case report. Subcutaneous phaeohyphomycosis due to Cladosporium cladosporioides. AB - A case of subcutaneous phaeohyphoymycosis caused by Cladosporium cladosporioides in a 25-year-old male is described. The clinical presentation was an elevated scaly suppurating lesion with sinuses on the right leg. The lesion healed completely with oral fluconazole therapy. The present report constitutes the first record of cutaneous or subcutaneous infection due to Cl. cladosporioides from India. PMID- 10838856 TI - Case report. Onychomycosis due to Chaetomium globosum successfully treated with itraconazole. AB - We have experienced a case of toenail infection caused by Chaetomium globosum which we treated with itraconazole 100 mg day-1 for 6 months, after which time the nail lesions were almost cured. Our case is the first reported case of onychomycosis caused by Ch. globosum in Japan, and the seventh in the world. PMID- 10838857 TI - Case Reports. Pityriasis amiantacea as manifestation of tinea capitis due to Microsporum canis. AB - We report three patients, two infants and an adult, in whom their tinea capitis clinically appeared as pityriasis amiantacea. The mycological studies showed infection due to Microsporum canis in all cases. Correct diagnosis as well as adequate antimycotic chemotherapy of this atypical manifestation of tinea capitis remain a challenge to the clinician. Epidemiological aspects must be regarded also. PMID- 10838858 TI - [Orthodontics in the 21st century. Authoritative conference 21 May 1998]. AB - The lecturer develops his ideas on the future of the special field emphasizing the importance that communications, perfection of new techniques and the realization of a more complete diagnosis concerning the human being as a whole, will assume. PMID- 10838859 TI - [Analysis of the development of the arch form after treatment using "ARCAD'Image" software]. AB - The dental arch post-therapeutic modification plays a significant role in relapse phenomenon. This article describes the design work of the dental arches used as a base for the study of modifications that have arisen during orthodontic treatment then during the retention stage using pre-formed arch wires. ARCAD'Image software tries to design the patients dental arches by submitting a linear regression mathematical design on some characteristic landmarks of a photograph of the buccal impression. The designed dental arch can then be considered as being the closest/nearest to the patients morphology. This model enables the practitioner to bend the arch wire and is used as a base for the study of arch shape modifications. Our study shows that the term "relapse" appears to be used excessively; it would be more matter of evolution due to changes in neuro muscular balance along with facial aging. PMID- 10838860 TI - [The temporomandibular joint: recentering...some debate!]. AB - In comparison with hip pathology, the authors emphasize the importance of the biomechanical factor in the stomatognathic systems physiology and pathology. They denounce a syllogism which is set as a reference for the justification of orthodontics and dentistry. PMID- 10838861 TI - [Could mouth breathing lead to obstructive sleep apnea syndromes. A preliminary study]. AB - The aim of this preliminary work is to determine an easy method to diagnose "buccal breather" children and "nasal breather" children. Then, to establish a possible connection with the syndrome of obstructive sleep apnea. 22 children agreed to participate. Clinical, orthophonic, orthodontic, postural and polysomnographical exams have been carried out. The proposed clinical exam turns out to be a good means of diagnosing between buccal breathers and nasal breathers. The aerophonoscope reveals velar inadequacies in buccal breathers. The latter also present osseous discrepancies mainly in the mandible. The polysomnography reveals a higher apnea/hypopnea index and more agitated sleep in buccal breathers. Mandibular lowering movements are more frequent and similar to those of adults suffering from apnea. These elements similar to those encountered in adults suffering from apnea make us think that buccal breathing could be the origin of obstructive sleep apnea, several decades later. PMID- 10838862 TI - [Muscular equilibrium and orthognathic surgery. A preliminary electromyographic study]. AB - Few studies have been written about the effects of orthodontic preparation and of the consequences of surgical movements on mandibular motricity. The aim of this preliminary report is to evaluate the modifications of muscular activity between the pre-surgical phase and its evolution during the year after surgery, together with the skeletal modifications found. The muscular activity of six patients with serious vertical discrepancies requiring rehabilitation of the lower facial height has been studied using electromyographical recordings. A longitudinal study of the EMG data has been established in order to discover a muscular adaptation method towards physiological equilibrium following the realization of a new facial diagram. The reduction of the lower facial height obtained through a surgical procedure including a maxillary impaction, appears to indicate that, at rest, the masseter muscles activity tends to normalize, whereas that of the temporal muscles may sometimes increase. During maximal contraction, facial hyper divergence reduction would lead to a sharp decrease in temporal and masseter muscular activity which would subsequently return to normal. The increase in vertical dimension would also cause significant changes in muscular tonus depending on the sagittal direction of the associated mandibular osteotomy. It would take longer to achieve muscular activity stability. These few results show: The existence of significant modifications following orthognathic surgery, possible reduction of high rest muscular activities, often revealing a cranio mandibular dysfunction, after vertical typology rehabilitation. The electromyographic recordings during treatment then enable to perfect the therapeutic re-evaluation of these major vertical discrepancies. PMID- 10838864 TI - [Neuro-occlusal rehabilitation and selective grinding: results after 1 year]. AB - The aim of this work is to illustrate the results, one year after selective grindings have been used in "Neuro-Occlusal Rehabilitation" (N.O.R.). These selective grindings have been described by Pr Planas and are used to free mandibular lateral movements, in order to facilitate correct development of the stomatognathic system. PMID- 10838863 TI - [Developmental tendencies of the sagittal tooth axis between 10-13 years of age and the end of life; impact on retention?]. AB - In this study, 854 lateral cephalograms of normal toothed subjects from 10 to 87 years old who had no orthodontic treatment are analyzed. The posterior angles established by molars and sagitally examined maxillo-mandibular incisor-canine groups during occlusion are measured with age. These modifications are quantified in a general manner and then depending on facial typology. PMID- 10838866 TI - [Variation of the position and inclination of incisors in the sagittal plane using 2 modes of retention]. AB - The evolution of the incisor position and inclination during retention depends on many factors. It specifically depends on: the amount of movements realized during treatment, the position obtained at the end of treatment, the type of retention used, the facial type, patient sex and age. PMID- 10838865 TI - [Recurrence: mechanisms and treatments]. AB - An original analysis by computer has been imagined. Etiological diagnosis in recurrence's cases is based on asymmetry. Different levels were investigated: dental arch, ridge, basal bone and mandibular position. Dental shift, deformed bone, eccentric mandibular and occlusal distortion were found. Each molar is independently moving. Mandibular basal bone is stabilized in the transverse dimension. Mandibular deviation induces compression or stretching in temporo mandibular articulation. Occlusal interferences involve distortion of cuspal plans. To reach stable objectives, molars require symmetrical position below one millimeter discrepancy, molar locking is to be had for the disto-vestibular cusp and ovoid arch. A special anchor apparatus allows to correct every wrong position of each individual molar, in each three-directional space. Mini-positioner or mini-splint perfect anterior misalignment. Sometimes, thermical memory archs are required. Ten fundamental rules that regulate occlusal and mandibular stability have been checked after treatment. PMID- 10838867 TI - [The French Society of Dento-facial Orthopedics. Commission of Terminology]. PMID- 10838868 TI - [The eclectic cellular tropism of cytomegalovirus]. PMID- 10838869 TI - [Autopsy today: an obsolete practice or an instrument for improving the quality of health care?]. AB - INTRODUCTION: Autopsies are a fundamental moment of clinical audit that have been progressively decaying. MATERIALS AND METHODS: The autoptic rates of 6 hospitals of Piedmont Region (1639 autopsies, years 1995-97) were calculated. The Positive Predictive Value and Sensitivity (indicators of concordance between clinical and autoptic diagnosis) were calculated for each diagnosis; all discrepancies were classified as type I (adverse impact on patient's survival) and type II (uncertain impact) clinical errors. The diagnostic performance of medical, surgical and emergency staffs were compared with one another. RESULTS: Autopsies are rarely performed on the patients who died in hospital (7.13%). Independently from the diagnosis, the global concordance expresses a Positive Predictive Value of 37.22% and a Sensitivity of 47.71%. 401 type I errors and 307 type II errors were found. The best performance is reached by emergency staff. AIM: To evaluate the use and the meaning of autopsies in today's Italian healthcare organisations. PMID- 10838870 TI - [Biomorphological features of "minimal" carcinomas selected from a series of 2077 breast cancer cases]. AB - Two hundred eighty nine pT1a/pT1b (less than < or = 1 cm in diameter) mammary carcinomas were selected from a series of 2077 consecutive cases of breast carcinoma. When compared with carcinomas of a larger size, they were significantly associated with a lower histological grade (SBR), a lower growth fraction (Ki-67 antigen in less than 20% of neoplastic cells) and a lower number of positive cases (more than 10% of neoplastic cells) as far as p53, c-erbB-2 oncoproteins and EGF-R, as detected by immunohistochemical methods, are concerned. Moreover, a significantly higher number of estrogen and progesterone receptors positive cases (more than 10% of positive cells), and a higher frequency of "other" histotypes with a favourable prognosis, was detected. No difference between pT1a and pT1b cases was noted so that "1 cm size" only stratifies a class with a better prognosis. It is possible that small cases, although invasive, did not undergone sufficient mytotic cycles to produce the sequence of genetic changes that characterize the "no return" phase of breast cancer. A 17% of pT1a and pT1b cases displayed lymph node metastases (mean 26 lymph node/case examined): the number is so high that cautions about simple lumpectomies and about sentinel lymphadenectomy, even in cases of small cancers, are necessary, until a larger number of studies will become available. PMID- 10838871 TI - [The anatomic tradition in Venice]. AB - Venice had a long tradition and great reputation in the study of anatomical science dating back to the 1300's. The "Serenissima" Republic favoured the study and practice of anatomy as part of medical professional formation. Before the construction of the anatomical theater of San Giacomo dell'Orio, which took place in 1671, anatomical dissections were performed in churches, convents, hospitals and private homes. Even though Venice was not a University seat, it boosted numerous Venetian anatomists, among whom Benedetti, Massa, Santorini, and the medical activity of illustrious professors at the nearby University of Padua such as Vesalio, Falloppio, Spigelio, Vislingio and Morgagni. PMID- 10838872 TI - [Hypertrophic and polypoid gastropathy associated with juvenile adenomatous polyposis coli]. AB - Juvenile Polyposis is a syndrome with gastrointestinal polyps and increased cancer risk. The commonest form of this syndrome is inherited as autosomal dominant trait and presents as Familial Juvenile Polyposis Coli. Another variant involves mainly the stomach and another is generalized throughout the gastrointestinal tract. We present the case of two brothers with polyposis coli complicated by colonic cancer. The polyps were of juvenile, adenomatous and mixed types. The two patients after a decade of colonic endoscopic polypectomies presented gastric involvement by polyps and needed multiple endoscopic gastric resections. One brother underwent total gastrectomy. This stomach showed diffuse polyposis of hyperplastic and fundic gland types within an unexpected background of foveolar and glandular hypertrophic gastropathy. The patients at present are followed up with endoscopic procedures. PMID- 10838873 TI - [Oncocytic meningioma. Case report]. AB - Among the histological variants of meningiomas the oncocytic subtype is rarely observed. Up-today, only six cases of oncocytic meningioma are described. This subtype of meningiomas shows an aggressive behavior and recurrences are more frequent. We describe a case of oncocytic meningioma in a 78-years-old woman. The patient had a history of breast cancer diagnosed 9 years before the brain biopsy; bilateral mastectomy and adjuvant chemotherapy was performed. She had a right frontal tumour measuring 3 cm in diameter. The patient is alive and well eleven months after surgery. The tumour was composed by large polygonal neoplastic cells with finely granular eosinophilic cytoplasm. Neoplastic cells were arranged in sheets and nests delimited by thin fibrous septa rich in vessels. Psammomatous bodies were also present. Mitoses were rare and necrosis was absent. Oncocytic differentiation was demonstrated by conventional histology and immunohistochemistry. Immunohistochemistry revealed a strong and diffuse positivity for antimitochondrial antiserum, vimentin and EMA; a focal reactivity for cytokeratin was observed. The rarity of oncocytic meningiomas is underlined with only six cases described in the world literature. The immunophenotypic profile and the differential diagnosis of the neoplasm is discussed and the concept of oncocytic meningioma as a distinct entity of tumour is emphasized. PMID- 10838874 TI - [Mucinous adenocarcinoma of the prostate: description of a case]. AB - INTRODUCTION: We report an occasional biopsy of primary mucinous adenocarcinoma of the prostate with review of the literature and discussion about all criteria used to classify this clinical-pathological entity. MATERIALS AND METHODS: Histochemical (Alcian Blue and P.A.S.) and immunohistochemical (P.A.P. and P.S.A.) stainings were performed. PMID- 10838875 TI - [Benign hemangiopericytoma of the bladder: description of a case and review of the literature]. AB - Hemangiopericytoma (HPC) of the viscera is very rare and only six of these tumors arising in the urinary bladder have been previously reported. The differentiation of HPC from other neoplasms with prominent vascular pattern may be a diagnostic problem. The difficulty is further enhanced by the lack of distinctive immunohistochemical features of this tumor. Here is presented a case of benign HPC occurring in the right lateral wall of the bladder of a 37-year-old woman. The mass is completely transurethrally resected. Histologically, the tumor is characterized by a monomorphic population of polygonal or spindle-shaped cells packed around branching vascular channels, with "staghorn" configuration. There are no foci of necrosis or hemorrhage. The mitotic count is 2 per 10 HPF. Immunohistochemically, the tumor cells react with anti-bodies against Vimentin. Cytokeratin, Actin HHF35, S-100, CD 34, Factor VIII-related antigen are not expressed within the lesion. Ultrastructurally, the tumor cells surround endothelium-lined vascular channels. There are pinocytotic vesicles on the cell borders. The cells are separated from one another by deposits of basal lamina like material and collagen. On the basis of clinical, histological, immunohistochemical and ultrastructural observations, we believe that the case reported is a primary benign HPC of the bladder. There is no evidence of recurrence or metastasis 32 months later. PMID- 10838876 TI - Chromatin texture by computer-assisted image analysis. PMID- 10838877 TI - [The bug in the brain: a quinquagenarian fantasy]. PMID- 10838878 TI - [Preparation technique for studying muscle biopsies]. PMID- 10838879 TI - [Soft-tissue neoplasms in children]. PMID- 10838880 TI - [Myofibroblastic tumors]. PMID- 10838881 TI - [Rhabdomyosarcoma]. PMID- 10838882 TI - [Ewing's sarcoma and related conditions]. PMID- 10838883 TI - [Undifferentiated sarcomas: role of electron microscopy in their differential diagnosis]. PMID- 10838884 TI - [Ultrastructural study of ancient human tissues]. PMID- 10838885 TI - [Handling of material, including paraffin-embedded specimens, for diagnosis by electron microscopy]. PMID- 10838886 TI - [Celiac disease or chylomicron retention disease?]. PMID- 10838887 TI - [Wolman's disease: a case with malabsorption and 2 cases with virus- negative fatty liver cirrhosis]. PMID- 10838888 TI - [Glomerulocystic kidney in advanced hemolytic-uremic syndrome]. PMID- 10838889 TI - [Intestinal stromal tumors with skenoid fibers in patients with type I neurofibromatosis: histological and ultrastructural evaluation of a case]. PMID- 10838890 TI - [Chordoid tumor of the 3d ventricle: immunohistochemical and ultrastructural study of 3 cases]. PMID- 10838891 TI - [HMB45-negative, clear-cell amelanotic melanoma with aberrant melanosomes]. PMID- 10838892 TI - [Small-cell osteosarcoma]. PMID- 10838893 TI - [Polymorphism of poorly differentiated rhinopharyngeal carcinoma: description of 3 cases]. PMID- 10838894 TI - [Diagnostic considerations in a case of poorly differentiated carcinoma of the mediastinum]. PMID- 10838895 TI - [What is it? Electron microscopy quiz]. PMID- 10838896 TI - [Economic expediency and medical efficiency of methods for active detection of tuberculosis]. AB - The data collected in the Novgorod region show a significant deterioration of the tuberculosis epidemiological situation. The economic and medical efficiencies of the current methods of active detection of tuberculosis are comparatively assessed. There is evidence for high effectiveness of enzyme immunoassay in detection of blood tuberculosis antibodies. A new concept of a differentiation system for of organization of goal-oriented prophylactic population surveys for tuberculosis. PMID- 10838897 TI - [Dynamics of major constituents of tuberculosis morbidity in the population of Novosibirsk region]. AB - After processing the dynamic series of tuberculosis morbidity in the population of the Novosibirsk region by the smooth component methods, two major constituents were defined in morbidity dynamics. In total, they make up about half the total variability of tuberculosis morbidity. The first constituent is due to socioeconomic causes that are common for the whole region. The second one shows rather evident 10-year fluctuations. The same 10-year fluctuations are observed in the dynamics of tuberculosis morbidity among the cattle of the Novosibirsk region by out-stripping that in the population in the northern and eastern areas by 1-3 years. PMID- 10838898 TI - [Tuberculosis morbidity in the Sverdlovsk region]. AB - Childhood and adolescence tuberculosis morbidity rates are higher in girls than in boys. Adult males fall ill 5.3 times more frequently than adult females. Males fall ill with destructive (6.1 times), bacillary (4.1 times), pulmonary tuberculosis (5.3 times) more frequent than females. Females experience extrapulmonary tuberculosis 1.6 times as high as males. Convicts and those under investigation are ascertained to have earlier forms of respiratory tuberculosis than in the general population. At the same time, the facilities of the RF Ministry of Justice do not record extrapulmonary tuberculosis. Goal-oriented work with these facilities has caused a reduction in tuberculosis morbidity in the whole Sverdlovsk region. PMID- 10838899 TI - [Tuberculosis morbidity in a male population by the climatic, geographic and administrative rural areas of Dagestan]. AB - The epidemiological features of tuberculosis among rural males in Daghestan were studied. Its mortality in males was found to greatly vary by climatic and geographic areas by the east-west and north-south axes. 1990 to 1997 were marked by a rise in tuberculosis morbidity by both ecological and administrative areas. The data are given in the paper. PMID- 10838900 TI - [Role of social status of new cases of tuberculosis in choosing a therapy regimen]. AB - Two hundred and sixty-seven new cases of pulmonary tuberculosis were examined to study their social characteristics of wishes in choosing a therapy regimen. It has been found that the males who are 30-59 years old, live in rural areas, unemployed, have a low education level, bad habits, and no family prefer to treat at a day-and-night hospital. Those who are 20-59 years, have secondary and higher education wish to treat at a day hospital and in a polyclinic. Pension-age individuals choose to treat at a day-and-night hospital nearly equally frequently. The patients stopped treating at a day-and-night hospital 2 times more frequently than those in a polyclinic or at a day hospital. PMID- 10838901 TI - [Magnetic laser therapy in combination with lymphotropic drugs administration in treating teenagers with common forms of tuberculosis]. AB - The paper shows the efficiency of combined use of magnetic laser therapy (MLT) and lymphotropic drug therapy in 40 teenagers with common forms of tuberculosis. The use of laser makes it possible to modify methods of regional lymphotropic therapy, to prolong its use and increase indications for it. MLT in combination with lymphotropic drug therapy accelerates positive changes and hence enhances the efficiency of tuberculosis treatment. PMID- 10838902 TI - [Humoral and cellular immunity in meningitis of various etiology]. AB - The parameters of humoral and cellular immunity were studied in patients with meningitis due to tuberculosis and other diseases. Peripheral blood and spinal fluid were measured in 91 and 76 patients with tuberculous meningitis, in 75 and 59 with purulent meningitis, and in 54 and 33 with serous meningitis, respectively. Comparing assessment of the findings revealed profound changes in the content of T lymphocytes and their subpopulations. With this, impaired cellular immunity appeared not only as changes in the relative count of immuno complement cells, but in their different functional activity detectable by blast cell formation. Thus, in tuberculous meningitis, 18.11 +/- 0.95% of peripheral lymphocytes and 15.1 +/- 1.2% spinal fluid were exposed to the specific blast cell transformation antigen PPD whereas they were 1.94 +/- 0.3 and 2.4 +/- 0.9% (p < 0.001) and 1.86 +/- 0.28 and 3.1 +/- 0.8% (p < 0.001), respectively. Elevated spinal fluid levels of beta 2-microglobulin and IgE in patients are shown to be a differential diagnostic sign typical of tuberculous meningitis. PMID- 10838903 TI - [Immunity parameters in children with prior intrathoracic tuberculosis]. AB - Fifty-four children who had experienced intrathoracic tuberculosis were clinically and immunologically studied 3-10 years following chemotherapy. The count of T cells and their functional activity were found to be normal in 90.7% of children. Tuberculosis antibodies were identified in 62% of children, which suggests only long preserved specific sensitization. The tuberculosis antigen titer 1:16 was detected in 5.5% of children with residual changes, which generates the necessity of examining such children for a latent active process. PMID- 10838904 TI - [Immediate outcomes of medical therapy in patients with caseous pneumonia]. AB - The immediate outcomes of treatment were studied in 119 patients with caseous pneumonia whose age was 20 to 64 years. In most cases, caseous pneumonia was found to preserve its classic traits by characterizing by acute onset (85.8%), prompt progression, high proportion of fatal outcomes in the early postoperative period (54.1% within 3 early postoperative months). There is an increase in cases of more "benign" course of this form of tuberculosis (subacute onset and prolonged slow progression) at the same time. The use of current methods for combined intensive antibiotic and pathogenetic therapies substantially improves the outcomes in this most severe group of patients, reduces the initiation of a phase of relative stabilization of a specific process and of cessation of bacterial isolation. PMID- 10838905 TI - [Manifestations of pulmonary tuberculosis concurrent with diabetes mellitus and various haptoglobin phenotypes]. AB - For diagnosis, detection of the specific manifestations of pulmonary tuberculosis (PT) concurrent with diabetes mellitus, 48 patients with insulin-dependent diabetes (IDD) and 132 with noninsulin-dependent diabetes (NIDD), who carry various haptoglobin (Hp) phenotypes were studied. It has been found that PT develops in IDD mainly in 5-10 years and in NIDD in 1-4 years. The gravest course of both types of diabetes is frequently encountered in those having Hp 2-2 phenotypes and slightly less frequently in those with Hp 1-1. The patients having these phenotypes have abnormalities in the levels of glycaric hemoglobin, 2.3 diphosphoglycerol phosphate, in the activity of the enzymes lactate dehydrogenase and glucose-6-phosphate dehydrogenase and acid-alkali imbalance. It is expedient to determine Hp phenotypes to evaluate the severity and prognosis and to choose a treatment policy for comorbidity and the proposed biochemical indices should be more widely used to evaluate carbohydrate metabolic disturbances in PT concurrent with diabetes mellitus. PMID- 10838906 TI - [Effectiveness of treatment in patients with pulmonary tuberculosis and chronic bronchitis by exposure on vertebro-osteo-sternal joints]. AB - A hundred and twenty six patients with infiltrative destructive pulmonary tuberculosis were examined. In 62 patients of them, the vertebroosteosternal joints were exposed as part of multimodality treatment in order to arrest an exacerbation of chronic bronchitis. A control group included 64 patients. The exposure of vertebroosteosternal joints was found to enhance the efficiency of treatment for chronic bronchitis in patients with pulmonary tuberculosis. This treatment promotes lower inflammatory intensity, diminished antigenemia and anti protein antibody production. To arrest lesions in the bronchial tree with the proposed package of methods also favours a higher efficiency of treatment of a tuberculous process. PMID- 10838907 TI - [Acute pancreatitis in patients with pulmonary tuberculosis]. AB - The data on 33 patients with pulmonary tuberculosis who had developed acute pancreatitis due to being treated with antituberculous drugs are analyzed. Emphasis is laid on the fact that all these patients had been treated for pancreatitis or suffered from diseases of the organs adjacent to the pancreas. This category of patients are recommended drugs consequently given from small to routine doses for 3-5 days to prevent the adverse pancreatic effect of antituberculous drugs. If there are signs of pancreatic failure, it is necessary to withdraw chemotherapy temporarily and to initiate pancreatic treatment. PMID- 10838908 TI - [Altered functional and biochemical parameters during treatment of patients with bronchial asthma with inhaled gaseous superoxide]. AB - To investigate the impact of adaptive oxidative training with inhaled gaseous superoxide (GS) on endogenous oxidative stress (EOS) and lung function in asthmatics and healthy volunteers, short-term GS inhalation was repeated. The study involved 27 patients (median age 42 (34 to 44) years with atopic bronchial asthma and a median disease duration of 130 (120 to 180) months prior to the investigation and 8 healthy volunteers whose median age was 20.5 (18 to 25 years). The rates of GS generation at a distance of 1 cm from the source was 0.25 mumol/min. The examinees inhaled GS nasally over 15 minutes per session, on an average of 20 times over 2 periods of 4 weeks each. Spirometric studies, a mathacholine challenge test, salbutamol test and blood cell count were performed, and blood antioxidative components were measured. There is evidence that GS inhalations in asthmatics cause an adaptive oxidative training and promote reductions in EOS, as well as activation of antiinflammatory mechanisms and improved spirometric parameters. PMID- 10838909 TI - [Immune defense in hormone-dependent bronchial asthma]. AB - The study was undertaken to examine the clinical and immunological features of hormone-dependent bronchial asthma. Long-term corticosteroid therapy was found to have a pronounced immunodepressive effect on both lymphocytic subpopulations and opsonophagocytosis. The sequelae of immunological disorders are lower anti infective defense, which contributes to a more severe course of the disease generally running in the presence of chronic obstructive bronchitis. The use of immunomodulators, extracorporeal techniques, which favours to reduce the adverse immunosuppressive effect of long-term therapy, is an alternative to corticosteroid dosage increment. PMID- 10838910 TI - [Comparative study of various diagnostic techniques efficiency for tuberculosis]. AB - Thirty-one patients with active pulmonary tuberculosis were examined. The efficiencies of modified luminescence microscopy (with superfine iron added), enzyme immunoassay (using two test systems made in Moscow and Saint Petersburg (Russia) and the MycoDot rapid diagnosis (USA) were compared. The highest sensitivity was found in the modified luminescence microscopy (45.2%) and enzyme immunoassay [Moscow test system (48.4%)]. The enzyme immunoassay (Saint Petersburg test system) and Mycodot were noted for its less sensitivity, but its high specificity. PMID- 10838911 TI - [Pre-seeding treatment of pathological materials at tuberculosis dispensary]. AB - For preceding treatment of pathological materials, the authors propose a decontaminating agent that contains plivasept at a concentration of 0.08%, the enzyme imozymase and sodium bicarbonate versus decontaminating agents containing chlorohexidine bigluconicum or plivasept alone. The authors' agent increases the proportion of Coch bacterial positive cultures and increases the informative value of luminescence microscopy by reducing the proportion of samples having single mycobacteria in the agent. The use of the proposed decontaminating agent saves the time of attending personnel by reducing the stages of preceding treatment of pathological materials. PMID- 10838912 TI - [Clinical features of concurrent respiratory tuberculosis and syphilis]. PMID- 10838913 TI - [Tuberculosis in HIV-infected patients]. PMID- 10838914 TI - [Complicated course of lymphatic system tuberculosis]. PMID- 10838915 TI - [A case of sarcoidosis in a patient with persistent granular Mycobacteria followed up over 18 years]. PMID- 10838916 TI - [Methodological aspects of management in phthisiology]. PMID- 10838917 TI - [Analytic review of dissertation studies in specialty 14.00.26 "Phthisiology" approved by the Higher Certifying Commission of the Russian Federation in 1998]. PMID- 10838918 TI - Variation of density, species composition and dominance of rotifers at a shallow tropical reservoir (Broa reservoir, SP, Brazil) in a short scale time. AB - Rotifers are the predominant group of zooplankton found in the majority of reservoirs, constituting more than 60% of the total zooplankton present. Being opportunistic organisms, they easily adapt to changes in environmental conditions, and it is therefore difficult to establish a constant level of composition and occurrence of a species in specific areas at specific times. In order to establish how changes in composition and predominance of a species occurs within a dynamic system, the daily variation in the total density of rotifers and that of the dominant species were studied at short intervals during a period of 20 days at Lobo (Broa) reservoir. Filinia pejleri and Keratella americana were the predominant species, making up 64.3% of the total rotifers. However, during the first five days of the study, there was a predominance of two genera, Conochilus (C. coenobasis and C. unicornis) and Keratella (K. americana and K. cochlearis), with the genus Conochilus being substituted by the genus Filinia from the seventh day onwards, which, together with the genus Keratella, was predominant until the end of the study period. The increase in density of Filinia pejleri and Keratella americana was found to be associated with increased wind speed. The resulting turbulence could cause changes in the behavior and metabolism of the organisms, increasing the rate of reproduction, which in turn could be attributed to the greater availability of food through the suspension of material from the lake bed. PMID- 10838919 TI - Preliminary assessment of heavy metal levels in Mytella falcata (Bivalvia, Mytilidae) from Bacanga River estuary, Sao Luis, state of Maranhao, northeastern Brazil. AB - The concentrations of Cu, Zn, Pb, and Cd were determined in the mussel Mytella falcata from the Bacanga River estuary, in order to evaluate the potential role of domestic effluents released "in natura" on the quality of the estuarine environment with consequences to shellfish resources. Cu concentrations ranged from 5.2 mg.g-1 to 13.1 mg.g-1, while Zn concentrations ranged from 49.1 mg.g-1 to 76.3 mg.g-1. Pb and Cd showed concentrations lower than 2 mg.g-1. The results showed low concentrations of Cu, Pb and Cd in the filter-feeding Mytella falcata in the study area in comparison with the maximum values stablished to foods by the Brazilian Government (Decree Law No 55,871). Finally, Zn presented slightly higher values than the maximum ones stablished by the Brazilian government. PMID- 10838920 TI - Reproductive ecology of the blue crab, Callinectes danae Smith, 1869 in the Conceicao Lagoon system, Santa Catarina Isle, Brazil. AB - Abundance of ovigerous females, size of the first gonadal maturation and the possible migration, route of the blue crab Callinectes danae from the Conceicao Lagoon system, Santa Catarina Isle, Brazil, are described. This lagoon is connected with the coastal area through a canal. A total of 1,124 crabs was caught during a 19 month sampling period. The reproduction and recruitment of juveniles occurred all year-round, with two peaks of abundance (February and September), correlated with the presence of ovigerous females (June and January). The mean carapace width at which the crabs attained gonadal maturity for the first time was 9.4 cm in males and 8.4 cm in females. The Conceicao Lagoon is a growth, reproduction, and spawning area for the species. However, egg eclosion occurs outside the lagoon following migration of ovigerous females to the open sea. After hatching the eggs, some females return to the lagoon, but males stay there for most of their life cycle. PMID- 10838921 TI - Foraging behavior of bee pollinators on the tropical weed Triumfetta semitriloba: flight distance and directionality. AB - We studied flight distance and directionality of bee pollinators on the tropical shrub weed Triumfetta semitriloba Jacq. (Tiliaceae), addressing (1) within- and between-plant movement pattern; (2) distances flown between plants; (3) flight directionality. Flowering plants were distributed in well-delimited clumps, in each of two pasture areas (A1 and A2) and one area of forest gap (A3), in Vicosa, southeastern Brazil. Five solitary bee species, Augochlorella michaelis, Augochloropsis cupreola, Pseudocentron paulistana, Ceratinula sp., Melissodes sexcincta, and two social bee, Plebeia droryana, P. cf. nigriceps were observed. All species moved mainly to the nearest flower on the same individual plant and, in between-plant movements, to the first or second nearest neighbor. All species moved non-randomly, presenting a flight directionality in departures (maintenance of flight direction), but with a high frequency of turn angles. It is suggested that this foraging behavior pattern occurred because of the resource quantity and quality (pollen or nectar), and environmental characteristics such as flower density and resource distribution. PMID- 10838922 TI - Clustering behavior of hermit crabs (Decapoda, Anomura) in an intertidal rocky shore at Sao Sebastiao, southeastern Brazil. AB - The clustering behavior and cluster composition of hermit crabs as well as the patterns of shell utilization of clustered and scattered individuals were studied. This study was conducted in the intertidal region of Grande Beach, Sao Sebastiao, southeastern Brazil. Samples were taken both in randomized transects and 1 m2 quadrats during low tide periods. Crabs were counted, measured (shield length), and sexed. Shells were identified and had their adequacy and condition (physical damage and incrustation) recorded. Clusters occurred mainly in air exposed areas and were dominated or composed only by Clibanarius antillensis. Other species like Paguristes tortugae, Pagurus criniticornis, and Calcinus tibicen were also present in these clusters, but in small numbers. Only one monospecific aggregation composed by individuals of P. criniticornis was recorded in tide pools. Almost all crabs were inactive, despite some that were submerged in tide pools. Most of the individuals of C. antillensis were clustered (70.88%). Scattered individuals were larger than clustered ones and occupied mainly shells of Tegula viridula, which seemed to be the most adequate shell to the crabs. Clustered individuals used less incrusted shells than isolated ones. In general, clustering in Grande Beach presented the same patterns of size and sex distribution, and shell utilization than others already studied, with the exception of the smaller cluster size registered in this area. PMID- 10838923 TI - Comparison of artificial diets for rearing Corcyra cephalonica (Stainton, 1865) (Lep., Pyralidae) for Trichogramma mass production. AB - The objective of this research was, based on biological studies, to determine and adequate diet for rearing Corcyra cephalonica (Stainton, 1865) in the laboratory so as to permit the rearing of this factitious host for Trichogramma mass production. The research was conducted at 25 +/- 1 degrees C, RH 60 +/- 10% and photophase of 14 hours. Six artificial diets were compared: a) whole wheat flour (48.5%), ground rice (48.5%) and sugar (3%); b) ground rice (97%) and sugar (3%); c) whole wheat flour (48.5%), rice flour (48.5%) an sugar (3%); d) whole wheat flour (97%) and yeast (3%); e) wheat germ (97%) and yeast (3%); f) rice bran (94%), sugar (3%) and yeast (3%); f) rice bran (94%), sugar (3%) and yeast (3%). All of the diets studied permitted the development of C. cephalonica although the diets with wheat germ and yeast and that consisting of rice bran, sugar and yeast proved to be the most adequate for rearing the moth. These diets reduced the total (egg-adult) cycle, shortened the egg laying period, and produced heavier adults. Studies on the fertility life tables showed that higher net reproduction rates (Ro) and finite ratio of increase (lambda) were obtained from adults reared on these diets. PMID- 10838924 TI - Low courtship song variation in south and southeastern Brazilian populations of Drosophila meridionalis (Diptera, Drosophilidae). AB - Drosophila meridionalis is a cactus-breeding species with a wide distribution in South America. Most populations of this species are geographically isolated, what provides a promising scenario for studying evolution. Former studies of this species revealed a remarkable karyotypic variation among its populations. Up to six distinct metaphase chromosomes were described, showing that this species is polymorphic at least at the chromosomal level. In order to elucidate the taxonomic status of populations showing different metaphase chromosomes, we analyzed the courtship song of five populations of D. meridionalis in South and Southeastern Brazil. In addition, we analyzed the metaphase chromosomes of each population. Our results show that, despite the two karyotype observed, most courtship song parameters did not vary among the populations. Altogether, our results suggest that D. meridionalis from South and Southeastern Brazil represents one species with an inter-population chromosomal variability. PMID- 10838925 TI - Factors affecting biochemical composition of seston in an eutrophic reservoir (Pampulha Reservoir, Belo Horizonte, MG). AB - Sestonic biochemical composition (lipids, proteins and carbohydrates) may change with varying environmental fluctuations. These changes and the effects in the nutritional status of algae consists of an actual increasing source of interest. The aims of this work were to establish the relationship between biochemical composition of seston and 1) a range of physical, chemical and climatological factors, and 2) the natural fluctuation in the species composition in phytoplankton assemblages of the Pampulha Reservoir, an urban eutrophic lake located in Belo Horizonte, MG. Seasonal changes in the biochemical composition were observed in this study. None of the considered abiotic factors alone seem to affect the biochemical composition. So the effects could only be understood by interactions among different environmental factors. On the other hand, the dominant algal groups probably have some influence in the observed changes in biochemical composition of seston. PMID- 10838926 TI - Variation and life strategies of the Trichoptera (Insecta) larvae community in a first order tributary of the Paquequer River, southeastern Brazil. AB - The Trichoptera fauna was sampled in a first order tributary of Paquequer River, in the district of Teresopolis, Rio de Janerio State. Quantitative samples of litter from pools, litter from riffles, sand, and stones were taken on each season from February 1991 to February 1992. The life cycles of common genera were categorized into three groups of biological strategies. The largest group represented the general profile of the community, in which ultimate instar larvae were predominant in spring and the younger ones in summer. Probably, emergence and oviposition occur in spring. PMID- 10838927 TI - Population dynamics and net primary production of the aquatic macrophyte Nymphaea rudgeana C. F. Mey in a lotic environment of the Itanhaem River basin (SP, Brazil). AB - In this paper we evaluated the population dynamics and obtained estimates of the net primary production of the aquatic macrophyte Nymphaea rudgeana in an arm of the Itanhaem River (Sao Paulo State, Brazil). This species presents, in the studied area, a broad seasonal variation of biomass. As from November (13.1 g DW/m2) we observed a gradual increase of biomass that reached a maximum in February (163.1 g DW/m2). Then, the biomass decreased, maintaining low levels until a new growth period. The reduction of biomass is associated to the development of floating aquatic macrophytes (Pistia stratiotes and Salvinia molesta) and, subsequently to environmental factors (higher salinity values) that are unfavorable to their development. The net primary production of N. rudgeana was estimated from the biomass data, and the annual productivity value was estimated between 3.02 and 3.82 t/ha/year. PMID- 10838928 TI - Dental development and ontogenetic diet shifts of Roeboides paranensis pignalberi (osteichthyes, Characinae) in pools of the upper Rio Parana floodplain (state of Parana, Brazil). AB - Species of the characid genus Roeboides are known for their habit of tearing off and ingesting scales from other fishes. Specimens of Roeboides paranensis were taken monthly from March 1992 through February 1993 in five pools of the upper Rio Parana floodplain and in one site in the Rio Parana itself, with the aim of relating the dental development to ontogenetic diet shifts. Between 15-22 mm SL, fish had teeth with hypertrophied bases (mammiform) that moved to the outside of the mouth in both maxillas. During ontogeny, the diet shifted, with the smallest individuals eating more microcrustaceans and the larger ones eating more scales. Lepidophagy is non-obligatory, because the ingestion of insects and other invertebrates occurred at all sizes. Spatial variation in diet was large, however diet similarity was great for Roeboides from three floodplain pools that had similar proportions of the main diet categories. PMID- 10838929 TI - Comparative study of the zooplankton composition of six lacustrine ecosystems in central Brazil during the dry season. AB - Zooplankton community from six lacustrine ecosystems located in Federal District (Central Brazil) was studied based on samples collected during the dry season (July to September). A total of 71 taxa were recorded: 44 rotifers, 17 cladocerans and 10 copepods. The highest number of zooplankton species was recorded in oligotrophic Bonita Pond (32 species) and the lowest number in hypertrophic waste stabilisation ponds (7 species). This tendency of decreasing the diversity with increasing trophic level was consistent with a cluster analysis of the samples based on Sorensen index of similarity. From the overall similarity dendrogram, two groups of ecosystems were distinguished: one containing the natural ponds Bonita and Formosa and the other comprising the reservoirs Santa Maria, Descoberto and Paranoa. The role of morphometric features in determining the zooplankton community in such lacustrine ecosystems was also discussed. PMID- 10838930 TI - Growth of Nile tilapia Oreochromis niloticus fed with different levels of alcohol yeast. AB - Two hundred and forty 45-day-old fingerlings of Nile tilapia (Oreochromis niloticus), sexually reverted with an initial average weight of 1.25 +/- 0.14 g, distributed in a totally casualized delineation, during 330 days. The effect of the substitution of 10%, 20% and 30% of the ration by distillery yeast (Saccharomiyces cerevisae) in balanced experimental rations on the development in the breeding of tilapias did not show a harmful effect up to the maximum tested level of 30%, showing that the choice of the yeast in the ration for these fishes depends on the availability and occasional cost. The yeast provides fish with good resistance to unfavorable environmental conditions, which could be verified since there were no statistical differences among the treatments with different levels of yeast and the witness. The long growth rate in terms of length and weight can be attributed to the little available space for each specimen, not obeying the limit of populational density, as well as reduction of food ingestion, which coincide with the winter season, above of the absence of natural feeding. PMID- 10838931 TI - Antennal malformations in light ocelli drones of Apis mellifera (Hymenoptera, Apidae). AB - Malformed antennae of Apis mellifera light ocelli drones were drawn, dissected and mounted permanently on slides containing Canada balsam, in order to count the olfactory discs present in each segment, in comparison with the number of those structures in normal antennae of their brothers. Some drones presented morphological abnormalities in a single segment of the right or left antenna, but others had two or more malformed segments in a same antenna. Drones with malformations in both antennae were also observed. The 4th and 5th flagellum segments were the most frequently affected. In a low number of cases the frequency of olfactory discs in malformed segments did not differ from that one recorded for normal segments. However, in most cases studied, the antennal malformations brought about a significant reduction in the number of olfactory discs from malformed segments. PMID- 10838932 TI - Reproductive phenological patterns of cerrado plant species at the Pe-de-Gigante Reserve (Santa Rita do Passa Quatro, SP, Brazil): a comparison between the herbaceous and woody floras. AB - A natural vegetation area, with 1269 ha, composed mainly by cerrado, located at Santa Rita do Passa Quatro Municipality, Sao Paulo State, southeastern Brazil (21 degrees 36-39'S, 47 degrees 36-38'W), was studied. From September 1995 to February 1997, a floristic survey was carried out, when all vascular plant species at reproductive stage were collected. The flowering and fruiting patterns of the community were studied, comparing the herbaceous and the woody species. In the herbaceous component, 239 species were found and, in the woody one, 108 species. The woody species flowered mainly at the beginning of the rainy season, while the herbaceous ones produced flowers generally at the end of that season. The proportion of anemo and autochorous species was greater in the herbaceous component. The zoochorous ones, on the contrary, were more frequent in the woody component. At the dry season, when their dispersion is more efficient, the proportion of anemo and autochorous species producing fruits was higher. During the rainy season, when their fruits become attractive for longer time, the zoochorous species fruited more intensely. PMID- 10838933 TI - [Variation of above-ground biomass of Allagoptera arenaria (Gomes) O. Kintze (Arecaceae) at a palm shrub community on the Marambaia beach ridge, Rio de Janeiro, Brazil]. AB - Variation of above-ground biomass of Allagoptera arenaria (Gomes) O. Kuntze (Arecaceae) along five topographic profiles perpendicular to the ocean was examined in a palm scrub community on Marambaia beach ridge, Rio de Janeiro State, Brazil. Aerial biomass was positively correlated with distance from the sea (F = 39.57; R2 = 0.69; P < 0.01) as was detritus cover (F = 525.92; R2 = 0.92; P < 0.01). A. arenaria growth is closely related to the topography of the beach area. Dense populations of this palm enrich the soil by increasing organic matter under the plants through dead leaf material. This promotes the accumulation of nutrients and the creation of micro-climates that favor the establishment of other species. PMID- 10838934 TI - Patterns of gall-forming in Ossaea confertiflora (Melastomataceae) by Lopesia brasiliensis (Diptera: Cecidomyiidae) in an area of Atlantic Rainforest in southeastern Brazil. AB - Patterns of galling by the gall midge Lopesia brasiliensis (Diptera: Cecidomyiidae) were studied in Ossaea confertiflora (Melastomataceae) in an Atlantic forest site at Ilha Grande, RJ. Out of the 81 plants surveyed, 55 (67.9%) bore galls. The number of galls per galled individual ranged from 1 to 261 and 94.4% of the galls were in leaves. The number of galls per galled leaf varied from 1 to 25. Total gall number was positively correlated with plant height. Larger and more ramified plants tended to have a smaller percentage of their leaves with galls and a lower density of galls per leaf than smaller plants. Plants that were close to other individuals of the same species tended to have more galls per leaf than relatively isolated plants. The observed patterns may be linked to strategies of optimization in the use of resources (i.e. oviposition sites) and predation avoidance by the gall midges. PMID- 10838935 TI - Histology of the trachea and lung of Siphonops annulatus (Amphibia, Gymnophiona). AB - The structure of the trachea and lung of Siphonops annulatus was studied in ten specimens of routinely fed animals. The trachea is constituted mainly by incomplete cartilage rings lined by a respiratory epithelium (ciliated and mucous cells) with variable morphology according to the region observed. A rich vascularization of this organ suggests its participation in blood-air gas exchange. The right lung in this species is developed and the left one is atrophied. This organ is constituted mainly by longitudinal septa formed by connective tissue, smooth muscle cells and blood capillaries. These structures are covered by pneumocytes of one type only, which present cytoplasmic particles that have been related with surfactant activity described in the lung of Gymnophiona. PMID- 10838936 TI - Biology of Corecoris dentiventris Berg, 1884 (Hemiptera, Coreidae) in tobacco culture (Nicotiana tabacum). AB - Corecoris dentiventris Berg, 1884 (HEM., Coreidae) represents a pest in the tobacco culture in Southern Brazil. Nymphs and adults cause the wilting and twisting of the tobacco leaves due to their sucking habit. There are very few works about this species and these are restricted to concise description of the adult morphology, records of host plants and enumeration of injuries. The present investigation aims to study the life cycle of this bug. The experiment was carried out in a tobacco culture (Virginia type, var. K 326) implanted in the experimental area of the Departamento de Fitossanidade of the Universidade Federal do Rio Grande do Sul, in Porto Alegre, RS. The life cycle was studied in field conditions, in nylon cages, from egg clusters oviposited by females of the colonizing generation. The preferred oviposition site was the main vein in the abaxial side of the leaf. The average incubation period was 13.7 +/- 0.04 days and the egg viability, 99.63%. The average periods of the different instars of the bug, in days, were: 5.4 +/- 0.08, 9.7 +/- 0.19, 4.5 +/- 0.08, 5.3 +/- 0.08, 9.0 +/- 0.13 respectively for the first, second, third, fourth and fifth instars. The first instar presented the highest mortality (13.86%). The accumulated mortality of the nymphal phase was 35.29%. The sex ratio was 0.97 males: 1 female. PMID- 10838937 TI - [10 years' of surgical practice in a free state]. PMID- 10838938 TI - [Subadventitial rupture of the internal carotid artery]. AB - The author describes the case of a 21-year-old patient who contracted an injury of the internal carotid by collision with the tip of a metal rod. The injury led to the development of a sub-adventitial rupture of the carotid artery and its subsequent thrombosis. The patient was admitted to the Clinic 12 hours after he lost consciousness. CT examination of the brain did not reveal malacia. The patient was operated--resection of the injured portion of the carotid artery was performed, carotid thrombectomy and reconstruction of the artery. After surgery the patient improved rapidly and gradually his condition returned to normal. PMID- 10838939 TI - [Percutaneous aspiration thrombectomy in the popliteal and calf area--a 4-year experience]. AB - PURPOSE: Retrospective analysis of results of percutaneous aspiration thrombectomy (PAT) in the popliteal and tibioperoneal arteries in patients with primary and iatrogenic peripheral embolization and during intraarterial thrombolysis. MATERIAL AND METHOD: Percutaneous aspiration thrombectomy was used in 41 patients (42 arteries). In 12 cases this method was used as the first choice in treatment of the peripheral artery disease, in 14 cases during intraarterial thrombolysis and 16 times after iatrogenic embolization secondary to balloon angioplasty. An antegrade approach to the common femoral artery was performed in all cases. In 40 patients the aspiration thrombectomy was performed from the popliteal region and proximal parts of tibioperoneal arteries. RESULTS: Overall success (n = 42) was achieved in 67%, partial success in 23%. Aspiration thrombectomy was unsuccessful in. 10%. PAT was successful in 88% after iatrogenic peripheral embolization (n = 16), in patients during thrombolysis (n = 14) a good result was achieved in 40%, partial success in 50% and this method failed in 10%. Primarily performed aspiration thrombectomy (n = 12) in occlusion of peripheral arteries was successful in 50%. No major complications were recorded. CONCLUSION: PAT is a highly effective method in treatment of iatrogenic peripheral embolizations after endovascular procedures and effective accessory method in combination with intraarterial thrombolysis. This method can be used primarily in acute occlusion of distal popliteal artery. PMID- 10838940 TI - [Treatment of esophageal achalasia]. AB - During the past years the authors performed at the First Surgical Clinic, Faculty Hospital Olomouc laparoscopic cardiomyotomy with Dor's plastic operation in 17 patients with achalasia of the oesophagus. From the above number 15 patients were relieved of their complaints, two patients were reoperated. The cause of failure of the operation was in one case previous treatment with botulotoxin, in the other case achalasia grade IV, refractory to this type of treatment. The authors consider laparoscopic oesophago-cardiomyotomy a safe procedure, effective in particular in the earlier stages of the disease. They warn against excessive protraction of surgical treatment, frequently caused by selection of ineffective non-surgical therapy. PMID- 10838941 TI - [Sentinel node biopsy in the staging of breast carcinoma. Radiation-guided surgery of sentinel nodes]. AB - During the period between January and August 1999 the authors performed in 26 patients with breast cancer stage T1 to T3, N0 to N1b preoperative identification and histological examination of the sentinel node as compared with examination of samples of the axillary lymph nodes. In the examined group the sentinel node and all other nodes were oncologically negative in 57.6% of cases, both were oncologically positive in 23% cases In 3.8% the sentinel node was oncologically positive and the other nodes were oncologically negative. In 15.4% it did not prove possible to identify the sentinel node. The authors did not record any case of false negativity of the sentinel node. In the authors opinion the use of the sentinel node for staging of breast cancer in early stages is a perspective method which makes it possible to eliminate standard dissection of the axilla and thus permanent sequelae for the patient. The authors are aware that the group of their patients is small but the results do not differ from those reported recently in the literature abroad. PMID- 10838942 TI - [Gastroplegia after elective intestinal surgery]. AB - The postoperative gastric atony presents a regular early complication after each laparotomy. The purpose of this study was to evaluate the impact of gastroduodenal decompression on the development of postoperative gastroplegia, the association of gastric atony with risk of other postoperative complications and the influence of nasogastric decompression on early postoperative feeding. 236 patients who underwent elective small bowel and colorectal surgery were evaluated according to the fact whether a prophylactic nasogastric tube was applied or not. Neither a higher incidence of gastric atony nor a rate of other postoperative complications in the patients with prophylactic gastroduodenal decompression was detected. The difference in the beginning of enteral feeding was statistically significant. The patients with gastroplegia had a higher incidence of the laparotomy wound dehiscence and of postoperative bronchopneumonia. The authors recommend a selective gastroduodenal decompression after elective bowel surgery only. PMID- 10838943 TI - [Carcinoma of the colon invading the duodenum and pancreas]. AB - In the submitted case-history the authors present the case of a 60-year-old woman with an extensive tumour of the transverse colon penetrating into the duodenum and head of the pancreas with an intimate relation to the upper mesenteric vein. An extensive curative en bloc resection included right-sided hemicolectomy, hemi pancreatoduodenectomy and partial parietal resection of the upper mesenteric vein. The pathologist identified in the resected tissue a medium differentiated adenocarcinoma of the large intestine damaging the head of the pancreas and infiltrating the duodenal wall. The lymph nodes were negative and no metastatic affections were revealed. The postoperative course was without serious complications. The few cases of this type reported in the literature illustrate the possibility of successful resection and thus a substantially improved prognosis of the affected patient. PMID- 10838944 TI - [Anastomosis using the Valtrac ring--pro and con]. AB - Authors present their experiences with the gastrointestinal anastomosis construction with the biofragmentable ring (Valtrac). Between May 1995 and June 1999 they used it in the group of 75 patients with mean age 58.4 (range 19-59) years. They used Valtrac most often--in 32 patients--to construct the anastomosis between small and large bowel after right hemicolectomy. One enterocutaneous fistula and one intestinal obstruction due to adhesions occurred in this group. Two patients had the signs of fecal impaction in postoperative period, which disappeared after the ring fragmentation. According to their experiences authors claim, that the anastomosis with Valtrac is a safe procedure with a few number of postoperative complications. Disadvantage in our conditions is its high price. PMID- 10838945 TI - [The syndrome of the common channel of the choledochal duct and the duct of Wirsung]. AB - The problem of the rare anomaly of pancreatic ad bile ducts--common channel syndrome is discussed. On the demonstrated case the necessity of ERCP in obscure etiology of chronic pancreatitis is suggested and the principle++ of surgical treatment--disconnection of the bile duct and the formation of choledochojejunoanastomosis on the excluded loop--is demonstrated which solved the problem. PMID- 10838946 TI - [Current views on pancreatic carcinoma]. AB - The modern methods of diagnosis can detect cancer of pancreas in stage in which possibility of operation is so high. The contribution of mini-invasive surgery is to prepare patient to operate on with success and low risk. We give an account of view to surgical problems with adenocarcinoma of exocrine pancreas which is developed in course of hundred years of surgical practice. We present retrospective study (not closed) of patients with that diagnosis during the last 5 years, complications, algorithm of diagnosis, procedures results. PMID- 10838947 TI - [Role of laparoscopic surgery in the treatment of abdominal pain in children]. AB - Principal indications of the coelioscopy in pediatric surgery is the laparoscopic appendectomy. In compare with classic techniques it has several advantages but as well as some limitations. During decision about eventual laparoscopic intervention in children is important to take into the consideration personal and material facilities of the unit or hospital for pediatric coelioscopic interventions, their accessibility as well as longer operating time and higher price of the operation. It is amoral to use the laparoscopic appendectomy as a method of teaching surgeon and team to the technique. The authors consider the laparoscopic exploration of the peritoneal cavity with subsequent appendectomy to be a very suitable technique for examination and treatment of pre-pubertal and pubertal girls or young women with chronic or repeated abdominal pain of the uncertain origin, however, its indication in indubitable acute appendicitis of a boy is questionable. PMID- 10838948 TI - [Significance and possibilities of cytostatic treatment of malignant tumors: testing the effects of cytostatics]. AB - The authors present an account on the initiation of a study concerned with the administration of cytostatics according to the sensitivity of the tumour cells. To assess the sensitivity the MTT test was used. The method is described in the paper. Four groups of tumours were examined by the MTT test: breast cancer, carcinoma of the colon and rectum, of the lungs and oesophagus + stomach. Six cytostatics were tested: 5-fluorouracil, cisplatinum, daunorubucin, paclitaxel, vincristine, and vepeside. Evaluation of the sensitivity of different tumours was based on the median of TCS50. The results of the MTT test were not used so far in the therapeutic protocol in all patients where the examination was made, as in solid tumours, contrary to haemo-blastomas, usually common empirically tested protocols are used. The authors reflect whether individually administered cytostatics according to the MTT test can improve the prognosis of patients with malignant diseases. They assume that long-term follow-up of the patients may provide favourable results in particular because more and more effective cytostatics are becoming available. PMID- 10838949 TI - [Iatrogenic injury of the pelvic artery during hip joint replacement and errors in the treatment of this complication]. AB - The authors present the case-history of a serious injury of the pelvic artery in a 70-year-old female patient during replacement of the hip joint, incl. some errors made in the treatment of this complication. The authors discuss medical and organizational aspects of the solution of this case. PMID- 10838950 TI - [The concept of surgery as a profession]. PMID- 10838951 TI - The value of ophthalmoscopy in the diagnosis of arterial hypertension in patients with ischemic brain disease. AB - Beside dyslipoproteinemia, one of the key risk factors for the onset of brain atherosclerosis, as well as ischemic brain disease (IBD) is arterial hypertension. Significant number of patients is not aware of their hypertension, and a paradoxical blood pressure decrease can occur at the onset of IBD, due to the failure of autoregulation mechanisms. Likewise, valid anamnestic data can not frequently be obtained due to difficulties in communication with patients. Regarding these facts, our hypothesis was that ophthalmoscopy in patients with IBD had the greatest sensitivity in the diagnosis of hypertensive disease, its duration and severity. For that reason, the purpose of this study was to determine the significance of ocular fundus examination in those patients with IBD who were not aware of their hypertension, or the high blood pressure was not registered at the admission. Study comprised 140 IBD patients selected upon the following criteria: ophthalmoscopy was performed by the same ophthalmologist, and IBD was diagnosed according to clinical criteria and by brain computerized tomography. Results of the study demonstrated that 26 (18.6%) patients, although not aware of having hypertensive disease, had grade I hypertonic fundus, 14 (10%) had grade II, and 8 (5.5%) had grade III hypertonic fundus, which indicated the high sensitivity of ophthalmoscopy in the diagnosis of hypertensive disease, as well as its duration and severity. This is particularly important in patients with negative history of hypertension, and also suggests the significance of routine ophthalmoscopy in normotensive patients. PMID- 10838952 TI - Brainstem auditory evoked potentials following head injury. AB - Brainstem auditory evoked potentials (BAEP) were examined in 40 patients with subjective disorders following closed head injury (CHI), with the established degree of recovery and performed CT-scan of the head. For all BAEP parameters the interval of normality was defined as 3 SD above and below mean value in the control group comprised of 20 healthy subjects. The upper limits of thus defined intervals of normality enabled the formation of four types of findings: type 1- normal finding that was registered in 23 (57.5%) patients; type 2 was a sum of individual findings with the prolonged interpeak latencies, but without the change of relative amplitude V:I--7 (17.5%) recordings; type 3--the findings where the fall of relative amplitude V:I was registered together with the prolongation of interpeak latency. It comprised of 4 (10%) recordings and the type 4 included 6 (15%) individual recordings with registered low RA V:I (0.8 or lower). The explanation of the most probable genesis of registered changes was presented. PMID- 10838953 TI - [Frequency, etiology, localization and surgical treatment of war injuries with tissue defects]. AB - The retrospective analysis of 1,514 cases treated at the Clinic for Plastic Surgery and Burns of the Military Medical Academy in the period between 1991 and 1995, established that the percentage of the injuries caused by gunshots and those caused by explosives during the conflict in former Yugoslavia, was more or less the same. The injuries caused by gunshot more often occurred on the head, neck, arms and trunk. The injuries of the legs caused by the explosives were more frequent, and they occurred in 83% of the cases. All the plastic surgeons who took part in the treatment of patients and in preparing the surgeons of other specialties for the treatment applied the original classification of the war injuries according to the structure of the defects that had occurred, to standardize the approach to the planning of treatment and the treatment itself of the wounded. In the delayed primary or secondary treatment of the injuries with the tissue defects all known plastic and reconstructive methods were applied. In the cases requiring the covering of the the tissue defect with the full thickness skin, local skin, fasciocutaneous, fascioadipose or muscle flap was chosen. Distant pedicled direct flaps were used in cases when it was not possible to use a more suitable reconstructive method. Free skin, myocutaneous or complex microvascular flaps were applied in cases of more extensive defects or if a more suitable solution could not be found. Our experience in surgical treatment of war injuries with skin defects during the civil war in former Yugoslavia has shown that over 50% of all the injured patients required the treatment of a plastic surgeon in a definite surgical treatment of a war injury. A multidisciplinary approach is necessary in the majority of the injured, and the surgical team is composed according to the affected area and the extent of the injury. PMID- 10838954 TI - [Sebaceous gland function in transplanted microvascular flaps]. AB - Sebaceous glands produce sebum that contains: triglycerides, esters of cholesterol, and sometimes even pure cholesterol, squalene and esters of wax. Microvascular transplantation exposed the tissue of free flaps to the temporary ischemia and anaerobic metabolism, inflammatory, denervative and reinnervative processes. The sebaceous glands of the free flaps were also exposed to the action of physiopathologic processes during the micro vascular transfer. Because of that, the function of the sebaceous glands of the transplanted free flaps was estimated compared to the function of the sebaceous glands of the recipients and donor's region environment. The investigation of the sebaceous glands of free flaps function has been performed in 32 wounded patients at the Clinic for Plastic Surgery and Burns, and Institute for Medical Biochemistry of the Military Medical Academy. All the wounded have undergone the microvascular transplantation of the cutaneous, myocutaneous and osteocutaneous flaps. Dry residue in either of the soluble substances was measured in the period from six to thirty six months after the transplantation. The obtained results indicated that the function of the sebaceous glands of the transferred flaps 25.36 months after the transplantation was weaker than the function of the sebaceous glands of the recipients and donor's region's environment and amounted to 98% of the function of the sebaceous glands of the recipient is region's environment. PMID- 10838955 TI - [Use of the internal thoracic artery in patients with stenosis of the left main branch of the coronary artery]. AB - Internal thoracic artery (ITA) is the graft of choice in myocardial revascularization. However, superiority of the ITA graft in patients (pts) with left main coronary artery disease is still a matter of debate. PATIENTS: In the period from November 1986 through February 1999, ITA graft was used for myocardial revascularization in 2860 pts. Stenosis of the left main stem was present in 229 patients (8.0%); there were 39 women (17.0%) and 26 diabetics (11.4%). Severely depressed left ventricular function was present in 32 pts (14.0%), while the average age was 56.4 +/- 7.2 years. RESULTS: Operative mortality was 1.3% and postoperative morbidity was 5.2%. Average postoperative hospital stay was 7.9 days (6-29). There were no differences in analyzed parameters between patients who received ITA graft and similar group of 240 patients who received vein grafts only, during the same period of time. All 16 patients operated on in the period 1986-1992 (6-12 years follow-up) are alive. CONCLUSION: It is safe to use ITA graft for myocardial revascularization in patients with left main coronary artery stenosis. Early operative results are favorable and these patients should not be denied the benefit of the ITA graft, since long-term results are proved to be good as well. PMID- 10838956 TI - [Significance of QT dispersion in the onset of ventricular arrhythmia in patients with heart failure]. AB - Prolonged QT dispersion which has been proposed as a marker of repolarisation inhomogeneity, may predispose to ventricular arrhythmias in a variety of cardiac disorders. The aim of this study was to compare some indices of QT dispersion in patients with heart failure compared to normal subjects. We have also tested the hypothesis that QT dispersion is a useful method for identifying the patients at high risk for ventricular arrhythmias. METHODS: There were 84 patients, divided into two groups. In the first group there were 62 patients with heart failure, in the sinus rhythm, while in second group there were 22 sex- and age-matched healthy subjects. Simultaneous 12-channel ECGs were recorded at a paper speed 50 mm/sec. Ventricular arrhythmias were quantified by 24-h Holter ECG and classified according to the Lown classification system. Only those patients with a class IVa, IVb, and V arrhythmia were considered to have complex ventricular premature contractions (PVCs). Measurements of QT, JT, and RR intervals were performed manually. Heart rate corrected QT and JT intervals (QTc and JTc) were calculated by Bazett's formula. RESULTS: RR intervals were similar in both groups (862 +/- 120 vs 840 +/- 86; ns). QT dispersion and rate corrected QT dispersion were significantly greater in heart failure patients than in controls (76 +/- 13 ms vs 37 +/- 11 ms and 89 +/- 21 ms vs 40 +/- 17 ms; p < 0.05). When, on the basis of the existing complex PVCs, heart failure patients were divided into two subgroups, QT dispersion and rate corrected QT dispersion were significantly greater in the subgroup with complex PVCs compared to patients without complex PVCs (84 +/- 14 ms vs 61 +/- 18 ms and 98 +/- 26 ms vs 66 +/- 21 ms; p < 0.05). CONCLUSION: All indices of QT dispersion were significantly higher in heart failure patients. QT dispersion is useful, noninvasive method for identifying heart failure patients at high risk for ventricular arrhythmia. PMID- 10838957 TI - [Characteristics of the use and abuse of psychoactive substances]. AB - The aim of the paper was to review the basic characteristics of drug abuse. Our questionnaire research included eighth-grade students of seven primary schools and first/second grade students of nine secondary schools in Kragujevac. In average, every fifth student took part in the investigation. Unfortunately, the research HAVE SHAWN that this phenomenon was wide-spread among adolescents; also, that their environment was well-aware of this but sadly, little was done to prevent the occurrence and spreading of drug abuse. Male adolescents, who usually show greater incidence of alcohol and cigarette consumption, also take some sort of drugs (52.9%) more frequently. Every thirtieth subject (or one student in each class, in average) used a psychoactive substance. The fact that every third participant knew somebody who took drugs implied that the phenomenon was spread among students. Commonly, they started taking drugs at the age of fourteen or fifteen, but every seventh or eighth of them did that so when he/she was about ten years old. It may be concluded, from the presented results, that the phenomenon is widespread, i.e. most frequently polytoxicomania. Also, the parents of the drug abusing students were frequently aware of the fact, but they responded in different ways. Regrettably, every fifth parent was not interested in the situation. Influence of all social subjects is crucial in prevention of this extremely important socio-medical problem, but the role of the family is essential and the hardest. The school and other subjects must also be included. PMID- 10838958 TI - [Importance of disposable medical materials and instruments in the prevention of intrahospital infections]. AB - Possibility for transmission of infectious diseases from patients to medical staff or vice versa is significant. Protective measures for patients as well as physicians must be applied and controlled. The aim was to investigate the microbiological status of disposable and reusable instruments and materials in order to establish the difference between them in protection from microorganisms. From the 122 samples of wet swab from reusables, 70.5% were bacteriologically negative and 29.5% positive. Seventy percent of isolated bacteria were a pathogens, and 30% were pathogens. All 80 samples of wet swab from disposable products were sterile. The frequency of bacteriologically negative samples among disposable products is highly significant (p < 0.01). These results confirm that the level of protection is higher if disposable products are used. Reusables should be replaced by disposable materials to decrease the incidence of nosocomial infections. PMID- 10838959 TI - [Importance of early detection of carcinoma of the large intestine]. PMID- 10838960 TI - [The most common viral infections after transplantation]. PMID- 10838961 TI - [Medical aspects of mobile cell phone use]. PMID- 10838962 TI - [When is "poor" left ventricular function really poor?]. PMID- 10838963 TI - [Immunologic and inflammatory aspects of atherosclerosis]. PMID- 10838964 TI - [Resection of post-intubation tracheal stenosis]. AB - The management of a patient with post-intubation circumferential stenosis and tracheostomy was presented. Tracheal resection and reconstruction were performed. The length of resection was four centimeters. The tracheostomy was included in the resected part. The problems of tracheal dissection and mobilization and approximation of the tracheal ends due to the anastomosis were presented. The possibilities of the intubation and adequate ventilation across the operative field were shown. PMID- 10838965 TI - [Myositis ossificans in recurrent synovial hemangioma of the knee]. AB - A case of myositis ossificans in 15-year old boy that occurred as a complication of the recurrent synovia hemangioma of the left knee was presented. The alteration was diagnosed as: osteosarcoma, chondrosarcoma and chondromyxoid fibroma by the pathologists from other institutions. The diagnosis of myositis ossificans was established using numerous histochemical and immunochemical methods (PAS, PAS diastasis, Alcian blue, Masson trichrom, von Kossa, Azur A, Toluidin blue, Goldner, method by Peris, Citokeratin, S-100, NSE and Vimentin). The presence of zonal phenomenon, as the one observed in the presented case, is one of the most significant criteria for differentiation of myositis ossificans from osteosarcoma. PMID- 10838966 TI - [The military physician, Dr. Zivojin Stojadinovic, author of the first Yugoslav manual for clinical laboratory diagnosis]. PMID- 10838967 TI - President's message. Governance of ANNA. PMID- 10838968 TI - Hearing the silence of patients who did not receive an anticipated organ transplant: "being second in line". AB - Admitting more than one potential organ recipient for a single available kidney is a common practice, resulting in one patient leaving the hospital without an anticipated organ transplant. The purpose of this study was to understand the life-world of end stage renal disease (ESRD) patients and their family members who experienced the phenomenon of "being second in line" for an anticipated renal transplant. After informed consent was given, indepth, face-to-face interviews were conducted with ESRD patients or family members. Interviews with 18 ESRD patients or family members who had experienced the phenomenon of leaving the hospital without an anticipated organ transplant were included in this study. The study was conducted in an mid-South university-based urban clinic that provided a variety of treatment services for dialysis and transplant patients. The narratives were tape recorded and transcribed verbatim by the researchers or a trained medical secretary. Hermeneutical analysis was used to bring forward themes found in these narratives. These narratives allowed patients and families to describe for themselves the lived experience of this event. Themes from the narratives included (a) knowing and not knowing; (b) having high hopes for a life without dialysis; (c) wanting the transplant, but not at the expense of someone else; and (d) having no voice for your experience. The final theme presented itself in three ways: (a) silencing the experience, (b) reframing by others as a "learning experience," and (c) reframing by others as a "misunderstanding." Data from this study do not support stopping the procedure of notifying more than one potential organ recipient about an available kidney, only that this is an important and meaningful experience unique to ESRD patients and their families. PMID- 10838969 TI - Acquired cystic kidney disease in ESRD. AB - Acquired cystic kidney disease (ACKD) in patients undergoing dialysis has been associated with life-threatening complications. Potential malignant transformation of the cysts has prompted concern regarding the need to screen patients for ACKD. The assumed increased morbidity associated with malignant transformation has not been clearly documented in the literature and may require further long-term analysis to adequately assess. Many reports on ACKD refer to the risk for malignancy, but other complications associated with ACKD may also arise. The development of ACKD has been shown to increase with increasing time on dialysis. Patients on dialysis appear to have a longer life expectancy and, consequently, more will develop ACKD. Complications related to ACKD may increase, which bolster efforts to establish specific guidelines related to the screening and management of ACKD. Nephrology nurses should be aware of symptoms suggesting a complication related to ACKD and confer with the nephrologist regarding evaluation. This article will discuss implications and management related to an increasing incidence of ACKD. PMID- 10838970 TI - An analysis of body weight and hemodialysis adequacy based on the urea reduction ratio. AB - The purpose of this study was to evaluate inadequate hemodialysis risk in patients with body weights in the upper quartile of a sample distribution using the urea reduction ratio (URR). Using a case-control design and a simple random sample (n = 315) of hemodialysis (HD) patients, postdialysis weights were divided into quartiles based with a cut-off value at the upper quartile, which was equal to > 81 kilograms (kg). The dichotomous outcome, URR > 65% and URR < 65% constituted the classification for inadequate dialysis risk. The odds ratio (OR) was used to evaluate inadequate dialysis risk based on this outcome. A multivariate logistic regression model was used to adjust for confounding variables and validated for goodness of fit. Those in the > 81 kg group were given more dialysis session time in minutes and used higher efficiency dialyzers as indicated by the coefficient of ultrafiltration (KUf), yet were more likely to have URRs < 65% compared to other patients in the sample (p < 0.001). This finding persisted in the logistic regression model when simultaneously fitting both gender and dialysis session time. Patients weighing > 81 kg experienced an increased risk of inadequate dialysis when compared to all others (OR 4.02, 95% CI [confidence interval] 2.217-7.29). A postdialysis weight > 81 kg increased the risk of inadequate dialysis for patients in this sample. This effect was confounded by a dialysis time x weight interaction term. Also, women were found to have a lower inadequate dialysis risk compared to men after adjusting for weight classification. We, therefore, conclude that patients who weigh > 81 kg may experience inadequate dialysis despite longer, more efficient dialysis sessions. Longer dialysis sessions may benefit some patients, but the effect in larger patients may not be a uniform response. PMID- 10838971 TI - The relationship between dialysis adequacies and sleep problems in hemodialysis patients. AB - This research examined the relationship between the incidence of sleep problems and hemodialysis adequacy (Kt/V) in hemodialysis patients. In addition, this study identified demographic variables that may be related to sleep problems of hemodialysis patients. The research design was cross-sectional, descriptive, and correlational. A convenience sample (n = 50) included 25 male and 25 female hemodialysis subjects from a private, for profit ambulatory dialysis clinic. The primary researcher interviewed subjects using a questionnaire that included demographics and questions regarding sleep habits. Two significant results were found. For subjects in the study aged 65 and older, increased dialysis adequacies were associated with a decreased number of awakenings at night. Second, women, regardless of age and education, reported using more sleep medications than men. Overall, this study did not find a relationship between sleep problems and dialysis adequacies. Further research is needed to increase awareness and understanding of the complexities of sleep problems in renal patients. PMID- 10838972 TI - Limitations of ferritin as a marker of anemia in end stage renal disease. AB - Management of anemia in patients with end stage renal disease (ESRD) requires careful monitoring of iron status. Although serum ferritin is used as an indicator of iron stores, its value is limited because ferritin is an acute-phase reactant; its level increases sharply in the presence of inflammation and infection. This article presents information on the value and limitations of serum ferritin as a marker of storage iron in healthy individuals and in patients with ESRD. It also discusses the need to consider the percentage of transferrin saturation, total iron-binding capacity, and immature reticulocyte fraction as markers of iron storage, particularly in patients with infection and inflammation. PMID- 10838973 TI - Aluminum toxicity in the 1990s. PMID- 10838974 TI - Methicillin-resistant staphylococcus aureus infection. PMID- 10838976 TI - The three Cs: change, conflict, and challenge. PMID- 10838975 TI - Managing anemia using laboratory trend analysis. Case study of the anemic patient. AB - Proper management of the anemia of end-stage renal disease (ESRD) requires chronic monitoring of an interrelated set of variables that can affect the erythrokinetic response. In most cases, therapeutic interventions should be determined on the basis of serial trends in laboratory values, thereby providing a historical pattern of clinical response. This article reviews the rationale for using laboratory trend analysis to manage anemia. A methodology for categorizing patterns in hemoglobin and hematocrit response to identify probable causes of hypo- or hyper-response to Epoetin alfa therapy is provided. PMID- 10838977 TI - Will nephrology nursing survive managed care? PMID- 10838978 TI - Manipulating survival and life quality outcomes in heart failure through disease state management. AB - Progressive DSM programs in the outpatient setting have positively influenced HF patient QOL as reflected in improved exercise tolerance and a reduction in emergency care and hospital admissions. For programs to be effective, pharmacologic and nonpharmacologic therapies must be incorporated. Nurses are active team members and can influence the success of the program, especially if they are knowledgeable and technically capable in management processes. Use of practice guidelines, standards of care, or algorithms allow for increased nurse autonomy and improve patient access to care and response times. The need to expand the DSM concept throughout the continuum of care is great. Future program development and outcomes research is necessary to provide direction in the implementation of high-quality, cost-effective programs. PMID- 10838979 TI - Evidence-based clinical outcome management in interventional cardiology. AB - In conclusion, through the use of pathways and case management, evidence-based clinical outcome management has occurred. The author's institution now has a process that enables it to accomplish three objectives: (1) the ability to track outcomes, (2) the ability to obtain information about opportunities for improvement and develop action plans for this, and (3) the ability to judge through continued variance analysis whether the actions taken made a difference or whether more changes are necessary. Based on this evidence, supported by the literature, the author's institution has been able to offer quality patient care at a reasonable cost. PMID- 10838980 TI - Rapid progression following cardiac surgery. AB - This article examines one institution's transition into rapid progression after cardiac surgery. This includes clinical guidelines and the integral use of outcomes measures to govern care. Fast-track initiatives are examined in the scope of cost containment. A comprehensive, multidisciplinary approach is emphasized throughout. PMID- 10838981 TI - Outcomes improvement following minimally invasive direct coronary artery bypass surgery. AB - The most progressive movement in the standardization of MIDCAB has occurred within the past 5 years. Standardization of care and continuous quality improvement are essential to improve outcomes and reduce costs for MIDCAB. At the authors' institution, perioperative clinical outcomes demonstrated no significant differences among a traditional single-vessel CABG and a MIDCAB in myocardial infarction rates, reoperations for bleeding, and cerebrovascular accidents. Differences were found in new-onset atrial fibrillation, extubation in the operating room, required transfusions, length of stay in the critical care unit, and overall length of hospital stay. Optimal perioperative critical care recovery may result from an evidence-based approach in the design and delivery of patient care. Standardized nursing interventions may be designed to improve efficiency and reduce inappropriate variations in perioperative care. Because MIDCAB is a palliative intervention for single-vessel CAD, multifactorial CRF management is a necessary adjunct for the achievement of optimal long-term outcomes. CRFs must be managed to maintain long-term arterial conduit patency rates (e.g., 20 y) and to prevent further progression of CAD in the native coronary arteries for MIDCAB patients. Nurse-managed, physician-directed CRF management programs are the avenue to provide such services. The reported clinical outcomes are appropriate variables to track for continuous quality improvement. These clinical outcomes are meaningful, measurable, and appropriate for evaluating the effectiveness of care but do not address quality of life, patient satisfaction, and efficacy of pain management. Nursing care must continue to evolve as more data become available. PMID- 10838983 TI - Management strategies for improving outcome following severe head injury. AB - Severe head injury is a major cause of disability, death, and economic cost. Significant reductions in mortality and morbidity can be achieved in patients with severe head injury through the use of evidence-based protocols and guidelines. Although no set standard is available for the management of patients with severe head injury, improved outcomes following severe head injury are seen with complete and rapid physiologic resuscitation and specific strategies that decrease ICP. PMID- 10838982 TI - Unplugging the mystery of carotid endarterectomy patient care. AB - The first two goals of health care must always be quality care and achievement of patient outcomes. In today's health care environment, these goals are achieved with an eye on the financial picture. Cost-saving efforts by decreasing LOS, decreasing the use of ICUs, and lowering laboratory and radiologic expenses without affecting the quality of care are requirements in today's setting. The process of creating a clinical pathway for patients undergoing CEA can help to examine your care and determine evidence-based practice. PMID- 10838984 TI - Early enteral feeding after surgery. AB - The GI tract is an important line of defense in the body's immunologic system. Without the proper care and treatment, this defense fails, rapidly increasing critically ill patients' susceptibility to added infections and system failures. Through the initiation of enteral nutrition, the GI tract can be preserved, and thus possible avoidance of some of the complications of critical illness may occur. A review of the research has demonstrated that the initiation of enteral nutrition in critically ill or postoperative patients is feasible and cost effective. As more research is completed, the role of the GI tract in critical illness and measures to maintain its integrity will be identified. In the meantime, the initiation of enteral nutrition is an important initiative. PMID- 10838985 TI - Management of patients with acute respiratory distress syndrome. AB - Acute respiratory distress syndrome is a complex clinical syndrome of respiratory failure that presents a challenge to every critical care team. Since the first clear description by Ashbaugh et al more than 30 years ago, much has been learned about the pathophysiologic process that occurs within the lungs after they suffer either a direct or indirect injury. Unfortunately, little success has been achieved in improving outcomes; however, hope is on the horizon. Current research evaluating optimal ventilator management, ECMO, the use of inhaled nitric oxide, and other experimental management strategies will hopefully combine to produce improved outcomes. PMID- 10838986 TI - Successful outcomes for the ventilator-dependent patient. AB - The challenges of working with the ongoing ventilator-dependent population are many. All have survived some element of critical illness to arrive at the point of step-down care. The team challenge is to craft a plan of care unique to their needs. Working through patients' pain of loss of control, the team helps patients and their families accept what is ahead--finding meaning in the smallest accomplishments and making peace. If the patient is to return home, achieving quality of life for whatever time is left is important. The ventilator dependent individual is the outlier, the variance, and the exception. Such is the challenge for the team caring for the ventilator-dependent population. It is our opportunity to make a difference. PMID- 10838987 TI - Developing an emergency department team for treatment of stroke with recombinant tissue plasminogen activator. AB - The EMS, ED, neurology, radiology, nursing, laboratory, and pharmacy departments are integral parts of the stroke team system. These are the key departments responsible for the support of the stroke team, or nucleus, which emanates from the departments of neurology and emergency medicine in the author's medical center. In the same way that cardiac victims are treated for "heart attack," so stroke victims should be treated for "brain attack." The emphasis on public awareness to community outreach and senior citizens groups through the aid of the public affairs department of the institution, the American Heart Association, and the National Stroke Association (NSA) has been a tremendous lift in this, the decade of the brain. The Public Broadcasting System and other television channels have helped enormously in publicizing stroke signs and symptoms, new treatment of stroke, and concurrent risk factors. New clinical research for thrombolytics and neuro-protective agents are now in progress to determine the best treatment for the damaged brain. The emphasis is changing: Time is brain. We hope to change the expenditure attributable to rehabilitation and managed care after stroke to the more hopeful prospects of prevention, early treatment, and fast recovery. We can then see that our friends and family members, as victims of stroke, may proceed to independence rather than long-term care facilities or nursing homes. This will be possible only if hospitals and medical centers nationwide adequately gear themselves for the treatment of ischemic stroke through the creation of a core stroke team, finely-honed interdepartmental cooperation, and the development of an efficient ED team that is fully immersed in the ethics and protocol of "brain attack." PMID- 10838988 TI - The effect of the clinical nurse specialist on patient outcomes. AB - The review of the literature shows a beginning body of evidence to support the positive effects of CNSs on patient outcomes. The most commonly examined patient outcomes were length of stay, complications, use of health care services, cost, and mortality rates. The study done by the author shows that patients with TKR operations on units with CNSs had better patient outcomes than patients on units without CNSs. Patients on units with CNSs had an overall shorter TLOS and fewer complications than did patients on units without CNSs, which is consistent with the literature. Health care providers are making continuing efforts to decrease operating costs. Because of economic pressures on health care administrators, a need to examine various job positions exists. Decisions regarding which positions are retained or eliminated should be based on well-designed research data. The effect of CNSs on patient outcomes can mean improved quality of care and cost reduction. As more research is conducted to show the effectiveness of CNSs on patient outcomes, health care administrators and consumers may become more convinced that such a person is a valuable member to have on the health care team. PMID- 10838989 TI - Implementation of a nurse practitioner role in an acute care setting. AB - During the implementation, the authors strived to clearly identify a person to focus on patient outcomes. Thus, they limited the ARNP's involvement in central functions and direct management of the staff. The overall implementation of the demonstration project has benefited patients, staff, and the health care team. The continuity provided allows the patient and family to interact with a consistent person. The ARNP functions as the key to directing patient care in a holistic manner while facilitating staff development. The demonstration project has given the authors an opportunity to evaluate the management structure and redefine roles to achieve those outcomes in the management arena. PMID- 10838990 TI - Outcomes-based education in a critical care nursing course. AB - Educational institutions are being asked to develop more meaningful ways to measure student learning. The critical care nursing faculty at this baccalaureate institution have delineated a process of outcomes-based education that they use as evidence of successful learning in their course. The faculty regard this as an ongoing project in which this evidence can only lead to continuing improvement. PMID- 10838991 TI - Pressure support ventilation: reducing the work of breathing during weaning. AB - Pressure support ventilation decreases the work of breathing by providing the patient with positive airway pressure during the inspiratory phase. The use of this type of ventilatory support is likely to increase over the next few years for patients, especially during the weaning period. By understanding how pressure support ventilation works and what patient parameters need to be monitored, the critical care nurse can help patients decrease respiratory muscle fatigue during weaning and thus decrease the weaning time for these patients. PMID- 10838993 TI - Tobacco withdrawal in CCU patients. AB - This project examined the hypothesis that smokers, undergoing forced abstinence from tobacco in the Cardiac Care Unit (CCU) setting, would have higher anxiety and more withdrawal symptoms than nonsmoking patients. The investigators found that anxiety was not higher in smokers than in nonsmokers, but that smokers did have more psychological withdrawal symptoms on the first day after admission. This article provides the critical care nurse with guidelines to assess patients for nicotine withdrawal symptoms, and offers suggestions for nursing care of these patients. PMID- 10838992 TI - Non-pharmacologic pain control for the CABG patient. AB - CABG patients have unique anatomical and physiological causes for postoperative pain that provide the critical care nurse with the challenge of identifying the most effective interventions for pain control. This article analyzes research reports on non-pharmacologic pain control for CABG patients and recommends state of-the-art nursing interventions based on this body of research. PMID- 10838995 TI - Saying "goodbye". PMID- 10838994 TI - Radiofrequency catheter ablation--Part 1: Pre- and post-procedure nursing responsibilities. AB - Radiofrequency ablation as a treatment for supraventricular and ventricular tachycardias has increased in popularity as a safe and effective mode of therapy. The critical care nurse's role includes pre-procedure, peri-procedure, and post procedure observations and interventions that contribute to positive patient outcomes. PMID- 10838996 TI - Identifying opportunities to ask patients about their treatment wishes. AB - Here is a challenging ethical case study about a women who when faced with multiple complications wondered if she could go on. Without specific information on her wishes, an ethical situation ensued when she became unresponsive with combined cardiac, renal, respiratory, and infective complications from cardiac bypass surgery and chronic COPD. Part I describes this case submitted by DCCN readers and Part 2 provides a case analysis by an ethical consultant with suggestions for nurses in similar situations. PMID- 10838997 TI - The implantable cardioverter defibrillator: patient and family education. AB - Patients undergoing implantation of implantable cardioverter defibrillators (ICD) have many questions and concerns about the device and how it will affect their lives. Teaching them about the ICD to prepare them for discharge is a nursing challenge. PMID- 10838998 TI - Origins: international perspectives, then and now. AB - It was because a number of people took time to listen to patients and families facing mortal illness that the Hospice Movement has grown world-wide since it began in the 1960s. The addition of new skills in pain and symptom control, the understanding of the problems faced by families and the need for research and teaching has brought the old traditions in care and caring into the present day. It has shown that it can be relevant in many settings and cultures and in countries with widely different resources. PMID- 10838999 TI - Origins: an American perspective. AB - The birth of hospice in the United States was fostered by the work of Florence Wald, former Dean of the Yale School of Nursing. Her activities are emblematic of the dedication of many other hospice volunteers who made hospice a reality in the United States. Nurturer, humanitarian and visionary, we salute Florence Wald and the many others who have contributed to the change in how end-of-life care is rendered in the United States. Congratulations and well done. PMID- 10839000 TI - History of the National Hospice Organization. AB - The National Hospice Organization grew out of efforts by the founders of the earliest hospice programs in the United States to protect their emotional investments in hospice care, to advocate for hospice interests in Congress and other public policy forums, to define standards for the fledgling movement, and to provide education on the nuts and bolts of running hospice programs for others who were interested in starting hospices in communities from coast to coast. Unlike the model of St. Christopher's Hospice in England, which began as a free standing in-patient facility and later added home care services, most U.S. hospices started as home care-based programs, often largely manned by volunteers. Among the crucial issues that have dominated the work of NHO during its first 21 years were passage and maintenance of the Medicare hospice benefit, ideological battles over the hospice philosophy, and efforts to extend hospice care to other populations, such as people with AIDS. PMID- 10839002 TI - Spirituality. AB - At the present time, there is a widening search for spirituality as distinct from organized religion, particularly as it relates to well-being, wholeness, and healing. In both professional and lay contexts, spirituality has come to the forefront of public consciousness. The place of spirituality within the hospice movement is not unaffected by this shift in popular priorities. Once the prerogative of chaplains and clergy, the nurturing of spiritual journeys is now becoming a common concern. Experiencing sensory spirituality can provide both caregivers and those for whom they care a blessed respite for bodies, minds, and spirits. PMID- 10839001 TI - Symptom control in hospice--state of the art. AB - There are a myriad of physical symptoms which can complicate the care of patients with advanced disease. Without knowledge of and attention to these distressing symptoms, the rest of the work of the interdisciplinary hospice team is greatly hampered. In this article, we review the management of ten prevalent symptoms in hospice care and to identify areas of clinical investigation underway and point of future areas ripe for investigation. PMID- 10839003 TI - Access to care. AB - In the twenty years since the National Hospice Organization began, hospice has grown tremendously. However, it still only serves a small percentage of terminally ill patients. This is because access to hospice services is limited by various restrictions to care. These barriers to care include societal attitudes towards death, diversity issues, socioeconomic issues, and eligibility issues. In order to develop and serve more of the population, hospice agencies must be flexible, creative, and use ingenuity to bridge the gaps that occur for some terminally ill patients. PMID- 10839004 TI - End of life care and decision making: how far we have come, how far we have to go. AB - While enormous progress has been made in improving the quality of care and the decision-making process for patients at the end of life, as a society we still have far to go to ensure that dying patients and their families have a comfortable and dignified death. In particular, reexamination and reconfiguration of our current decision framework is essential as our elderly population with chronic disease and slowly fatal conditions expands. With less certain disease paths and more complex and ambiguous choices, the growth of this geriatric population challenges us to develop a broader conceptualization of end of life care planning, so that end of life considerations are integrated into a larger anticipatory framework addressing options and needs as patients gradually decline. Within this framework hospice becomes a natural, integrated option along a continuum of care planning, rather than an abrupt alternative at a late stage of illness. End of life care planning must positively anticipate a robust array of needs and concerns well beyond the dramatic decisions to withhold or withdraw life-prolonging technologies usually found in advance directives. To embrace this broader framework it is critical that primary care physicians as well as disease specialists receive training in fundamental aspects of both geriatric and palliative care. Professionals from both of these disciplines must share expertise with each other, and should collaborate in advocacy efforts to effectuate changes in the clinical, policy and legislative arenas. PMID- 10839005 TI - End-of-life care: challenges and opportunities for health care professionals. AB - The failings of the American Health Care System in meeting the comprehensive needs of the seriously and terminally ill have led to both professional and public efforts to improve end-of-life care. Following a discussion of the shortcomings of end-of-life in America, this article describes the goals and philosophy of palliative care, while highlighting current innovative programs in end-of-life needs and insure quality of life for patients and families experiencing incurable, progressive illness. Health care professionals are called to respond to the challenges and opportunities of end-of-life care as individual health care providers, as members of professions, and as members of interdisciplinary teams committed to improving the care of the dying in America. PMID- 10839006 TI - Hospice vs. palliative care. AB - A greater interest in end of life care has been emerging in the United States. A description of the evolution of hospice and palliative care is reviewed with issues such as studies and reports on the care of the dying, physician assisted suicide, medical education, the author's two hospital audit surveys and discussion of the challenges that face both hospice and palliative care. PMID- 10839007 TI - The NHO Medical Guidelines for Non-Cancer Disease and local medical review policy: hospice access for patients with diseases other than cancer. AB - For much of its history, hospice focused on problems related to malignant disease. Recently, however, non-cancer diagnoses such as congestive heart failure, emphysema and Alzheimer's disease have comprised an increasing proportion of hospice referrals. This paper details criteria published by NHO and adopted by the US Health Care Financing Administration for hospice eligibility for common non-cancer diagnoses. A provisional list of domains for documenting "evidence of rapid decline," by which patients with advanced disease who do not meet criteria can still be certified for the Medicare Hospice Benefit, is also outlined. PMID- 10839008 TI - Hospice care and palliative care: a perspective from experience. AB - The establishment of the first Department of Pain Medicine and Palliative Care in a Medical Center in the United States is noteworthy. Since the design of the Department integrates a full-functioning hospice program within it, that has both a dedicated inpatient unit and extensive home care program, this Department represents a milestone in the development of the hospice movement, with full interrelationship between palliative care and hospice care. This paper will explore this interrelationship, its implications, and some of the background. PMID- 10839009 TI - Putting patient and family voice back into measuring quality of care for the dying. AB - Quality of care and quality of life change substantially for those with a serious chronic illness and nearing the end of their lives. As one dies, life takes on new shape-values change and things once ignored become more important. Existing quality of care measures do not attend to the changes in priorities or to dimensions that acquire new significance (e.g., Spirituality and transcendence). An important impediment to addressing the inadequacies in the evidence base for palliative care, improving shortcomings of care, and holding institutions or health care systems accountable for the quality of care is the lack of valid and reliable measurement tools. In this article, an overview is presented of an ongoing research effort to develop measurement tools which will utilize the patient and family perspective to measure the quality of care. PMID- 10839010 TI - Documenting the impact of hospice. AB - Hospice care has had an impact at many levels--on individual patients and families, on the health care industry, and on society. However, no comprehensive body of evidence has been generated that documents the impact of hospice care in terms that are meaningful to competitors, referral sources, and consumers. In part, this is because of the many challenges for evaluating hospice care. This paper describes recent efforts in the documentation of the value of hospice which have focused on outcomes measurement by individual providers rather than on large scale studies. Several groups are working to develop reliable measurement tools, to support standardized measurement in large numbers of hospices, and to collect information for benchmarking and comparison. Measurement of the impact of hospice care will set standards for other providers of end-of-life care and will document the expertise and knowledge of hospice professionals. Once established as centers of excellence in care of the dying, hospices will be well positioned for whatever delivery models may evolve for end-of-life care. PMID- 10839011 TI - New initiatives transforming hospice care. AB - Hospice care has been successful in serving a large segment of the terminally ill population in the United States. This article addresses a number of significant trends that may impact the future of hospice care. It is proposed that as many as one-third of those who die will not be in a position to make use of any end-stage program of care. Of the remaining, some will have difficulty being served by hospices due to uncertain prognosis and continued efforts at curative treatment. New models of caring for chronically terminally ill persons are being developed and are reviewed. A clearer definition of who ought to be served by hospice programs is encouraged. PMID- 10839012 TI - Reflections on death in America. AB - The project on Death in America was established to promote a better understanding of the experience of dying and bereavement and by doing so help transform the culture surrounding death. The Faculty Scholars Program provides three year fellowships for projects that explore critical aspects of the care of the dying, for those who will become academic leaders on the issue, as well as, role models and mentors to future generations of health professionals. A Grants Program broad enough to cover every aspect of the culture of dying allocates funding for innovative projects. A new grants program is underway to support projects aimed at enhancing the role of the humanities in transforming the culture of death and dying in America. PMID- 10839013 TI - Personal reflections. AB - A hospice pioneer and veteran of 23 years reflects on the early days of the hospice movement in the United States. The political, social and economic forces, which shaped the industry, are addressed from a local state and national perspective. Current challenges are briefly referenced with a call for hospice leaders to recognize the need for and identify where hospice fits into the broad end of life discussions. PMID- 10839014 TI - Reflections on the history of occupational stress in hospice/palliative care. AB - The concept of hospice and palliative care emerged a quarter of a century ago out of recognition of the unmet needs of dying persons and the social issues of the 1960s and 1970s. The issues of the day included the sexual revolution; a questioning of social values; an increased awareness of death resulting from the murder of the Kennedy brothers and Martin Luther King and daily television exposure to deaths in the Vietnam War, feminism, consumerism, reclaiming a more humanized role in the birth process, and hence in the process of death. The history of the hospice movement and the stress experienced by staff is traced from the early developmental days through to the present. Initially there was sometimes a struggle to integrate the concepts of relief of physical symptoms with meeting the psychosocial and emotional needs of patients and families, caregivers were expected to sacrifice much of their personal life for work, emotional intensity was high and supports were developed to ease some of the stress experienced by caregivers. From the early days team stress and burnout have been issues of concern. In the 1980s issues involved establishing funding sources, dealing with the new crisis of AIDS, and dealing with the gap between the ideal and the real. In the 1990s the economic climate has escalated some of the tensions that have always existed as hospice attempts to position itself within mainstream care with diminishing fiscal resources. These are issues that confront us as we move into the next century. PMID- 10839015 TI - Hospice reminiscences and reflections--an 18 year personal and professional love affair. PMID- 10839016 TI - The moment of death: is hospice making a difference? AB - The moment of death was a compelling image and dominant concept through much of history. In recent years this term has become destabilized by technological advances and changes in clinical practice. Perhaps even more significantly, the meanings previously associated with the death-bed scene and the final breath have become increasingly marginalized. Hospice programs continue to demonstrate that enlightened and dedicated care can markedly reduce the suffering of terminally ill people and their families. The vast experience acquired by hospice programs, however, has not yet been translated into a vision of the moment of death and the death-bed scene for our times. Several reasons are identified for the limited interest and even more limited hospice-based research into the death-bed scene and the moment of death. Hospice programs could contribute much to our understanding of the final moments of life if this should ever become a priority. PMID- 10839017 TI - Jennifer Mills. Interview by Melissa Ganon. PMID- 10839018 TI - Collective bargaining and workplace advocacy. Three states present their views. PMID- 10839019 TI - How to choose the right first job. PMID- 10839020 TI - Pediatric nursing. PMID- 10839021 TI - What's new in the OR? A look at perioperative nursing. PMID- 10839022 TI - A career in critical care nursing. PMID- 10839023 TI - Emergency nursing. More than meets the eye. PMID- 10839024 TI - 6 tips on preparing for the NCLEX-RN exam. PMID- 10839025 TI - Nursing organizations. PMID- 10839026 TI - Preach what you practice. PMID- 10839027 TI - 'A leg to stand on': an existential-phenomenological analysis. AB - The analysis of a paradigm-case (a person with an injured leg, an autobiographical history related by Dr Sacks) presented here is an illustration of how existential-phenomenological analysis can be done. The aim of existential phenomenological research is to analyse and to describe some dimensions of being. The person with an injured leg appears 'to bescotomized'. Well then, 'being scotomized' can be analysed and described as a particular mode of 'being', even as a particular mode of 'being-ill'. 'Being' has been studied (in the philosophical tradition of Husserl, Heidegger, Merleau-Ponty and Sartre) as that existential movement that brings an individual, the 'self', towards the world, meanwhile 'affecting' the personal body, personal time and space, the objects in a person's life-world, the individual's life-world and, finally, coexistence. The study of 'being-ill' is the study of the particular way the self, the body, time and space, the objects, the life-world, and coexistence are affected when being ill. 'Being-scotomized' may appear as affecting: (i) the self, isolating the subject and leading to a personal disintegration; (ii) the body to which he maintains an ambiguous relationship: being a body and having a body, and not having a part of the body any more; (iii) time and space being vanished with the vanished leg. The subject is 'at' a nowhereness and 'at' a motionless time; there is no meaningful future; (iv) the things in the person's life-world, to which it appears impossible to give the 'right' sense; (v) the life-world, revealing itself as a noland, characterized by silence and motionlessness; (vi) the others in this life-world that appears no-man's-land, uninhabitable nearly by definition. Coexistence reveals not to be possible. PMID- 10839028 TI - The extended role of the nurse: the United Kingdom perspective. AB - There has been considerable discussion and debate in the United Kingdom regarding the role and function of the nurse. This article outlines the main tenets of the debate, namely the extended/expanded role and function, and gives a brief overview of the situation to date. Professional guidance from the United Kingdom Central Council for Nurses, Midwives and Health Visitors is explored and the implications this has for the nurse in the exercise of accountability. The legal implications of the nursing function in an expanded role capacity is discussed, in particular in relation to legal accountability, with reference to existing case law. Recommendations are made for the development of the expanded role. PMID- 10839029 TI - Older women's experience of living with chronic leg ulceration. AB - In this study the authors sought to gain insight into the lives of older women, to focus on the experience of living with leg ulcers and to explore women's views by talking to them about their experiences. Twelve English-speaking women aged over 70 years who had experienced leg ulceration for 3 years or more were interviewed. Analysis of the interview text revealed two overarching themes: (i) gaining and maintaining control over vulnerable limb(s); and (ii) lifestyle consequences of chronic leg ulceration and impaired mobility. These themes contained several subthemes including: (i) nagging pain; (ii) self-expertise and infection; (iii) leakage, smell and embarrassment; (iv) fighting for skin and limb integrity; (v) wearing non-preferred apparel; (vi) loneliness; and (vii) coping, determination and hope. The findings of this research show that elderly women who live with leg ulcers experience multiple consequences. While the everyday problems of living with an unhealed would are addressed by the community nurse, other more subtle consequences may be overlooked. Recognition of the complex and sometimes hidden concerns of these women could help to avert the sense of helplessness which currently exists. PMID- 10839030 TI - The patient's perceived caring needs: measuring the unmeasurable. AB - The aim of this study is to elucidate the phenomenon of the caring need in the life-world of the patient and, on the basis of this, to compare patients' perceived caring needs with the six areas of need contained in a new instrument for patient classification, the Oulu Patient Classification (OPC). A total of 75 patients were interviewed and the data were analysed by means of a phenomenological-hermeneutical method. Seventeen perceived caring needs emerged from the process of interpretation. The patients experienced themselves as an indivisible unit that contained existential/spiritual needs and desires. The patients' caring needs are constituted from their problems, needs and desires, where human desire for life, love and meaningfulness may, express itself as bodily, psychical and existential/spiritual needs. A comparison between the patients' perceived caring needs and the OPC shows that patients' existential/spiritual needs do not emerge clearly enough and that the instrument should be supplemented by a caring perspective. PMID- 10839031 TI - Self-efficacy in chronic illness: the juxtaposition of general and regimen specific efficacy. AB - Changes in lifestyle are difficult for most people but necessary for those with a chronic illness, for whom changes may involve, among other adjustments, learning new behaviours and/or modifying one's lifestyle. The ease with which such changes occur depends on the person's efficacy beliefs and outcome expectations. This paper will discuss the conceptual issues related to self-efficacy: general, domain, and specific. Examples will be drawn from the health-related behaviour changes required to manage diabetes and rheumatoid arthritis. For this paper, regimen-specific or task-specific behaviour refers to the multiple tasks that the person carries out for management of their chronic illness. Confounding the issue of perceived efficacy (general, domain or specific), is the fact that compliance with all aspects of a recommended self-care regimen will not necessarily result in metabolic control for the person with type 1 diabetes mellitus, weight loss for the person with type 2 diabetes mellitus, or pain control for the arthritic person. PMID- 10839032 TI - A tentative model for developing strategic and clinical nursing quality indicators: postoperative pain management. AB - The purpose of the study was to evaluate the usefulness of a tentative model, based on important aspects of surgical nursing care, for designing strategic and clinical quality indicators. Objective postoperative pain management was chosen for the model because it is a priority area in surgical nursing care. Items within a questionnaire were designed by using the tentative model as a base and by means of a literature review. The questionnaire, directed to clinical nurses (n = 233), was compiled to establish the validity and the usefulness of the indicators. Fourteen items were assessed as essential for achieving high quality outcomes in postoperative pain management (11 as realistic to carry out, and 13 as possible for nurses to influence) with mean scores > or = 4 (on a 5-point scale). The conclusion reached was that the tentative model combined with a literature search was found to be effective for designing items that might be useful as strategic and clinical indicators of quality in postoperative pain management. PMID- 10839033 TI - A story: nursing-damaged lives. AB - This paper presents a story that captures forever a 'difficult', 'horrible' but in many respects totally 'normal' nursing moment. It is a short story of only one person's reality. On that fateful night in which many lives were changed forever, there were, of course, many realities, all of which hold their own truth. This tale is offered in the spirit of sharing and in the hope that others may find it useful. I believe that 'story telling' allows us to revisit and review our practice. In doing so, stories facilitate the discovery of nursing knowledge and the self. Ultimately this contributes to the development of expert practice. Nursing stories, then, become an excellent medium for nursing inquiry, from both an academic and a clinical perspective. PMID- 10839034 TI - Improving health care through innovative practice. PMID- 10839035 TI - Luck: what the nurse should know about it and how it affects nursing situations. AB - Luck permeates every aspect of human behaviour. Thus, luck is an aspect of nursing care and client belief of which the nurse should be aware. Beliefs about luck will influence client compliance with recommendations for actions as well as influence actions the client selects in relation to health. Beliefs about luck will also influence actions the nurse may take when responding to clients. PMID- 10839036 TI - Life control of members of the Kainuu Martha Organization in Finland. AB - The purpose of this study was to describe the life control of members of the Kainuu Martha Organization (the Marthas), based on their written autobiographic material. The study material consisted of written thematized autobiographies of members of the organization (n = 41). The material was analysed using qualitative content analysis with concepts of the health portrait. The Marthas were classified into three groups (member, participating and leading) on the basis of their participation in Martha activities. According to the generational categorization of Roos, the member Marthas represented the Finnish generations of well-being, structural change and shortage. They did not participate actively in the Martha activities. Their feeling of life control was based on the sense life made to them. The member Marthas emphasized the meaning of family in their lives. The participating Marthas represented the generations of structural change and shortage. They participated in the Martha activities as observers or by attending the functions of their local organizations. Their feeling of life control was related to a favourable attitude towards life and concern for their human relations. The leading Marthas represented the generation of shortage and participated actively in the Martha activities. They believed in the continuity of the Martha Organization in Kainuu. They had a strong feeling of life control. This manifested as an enterprising spirit in everyday life, which included a skill to live life as it occurred. The lives of the leading Marthas were characterized by a favourable attitude, a feeling of responsibility and initiative. PMID- 10839037 TI - The manifestation and nursing management of agitation in institutionalised residents with dementia. AB - This participatory action research study is the first formal research study undertaken by nurses in a nursing development unit. The study emerged as the result of nurses brainstorming issues that they perceived to be problematic in their nursing practice and their unit environment. The nurses of the psycho geriatric nursing development unit, Wallsend Aged Care Facility, New South Wales, Australia identified that the management of agitated behaviour manifested by their severely demented clients was a major challenge in their practice. As a result, a pilot participatory action research study was designed to measure how agitation manifested, to measure the severity of agitation and to identify current nursing practices used to manage it and their outcome. Results of the study demonstrated that nurse actions triggered the majority of most highly rated episodes of agitated behaviour in this group of elderly residents, and that the majority of these nurse actions related to those involved in carrying out activities of daily living for the residents. PMID- 10839038 TI - Truth-telling versus hope: a dilemma in practice. AB - This paper presents a brief outline of the attributes and therapeutic nature of hope in the context of truth-telling. It is suggested that, in some circumstances, withholding the truth to protect hope can be considered a morally acceptable option when truth-telling has the potential to destroy hope's therapeutic effects. A clinical vignette that highlights how the practitioner can be confronted with a clash of principles is presented. The possible consequences of both truth-telling and the withholding of truth are discussed and the moral positions of the health-care professionals involved are examined. This article is intended to provide a framework for discussion/group work, particularly at undergraduate level. PMID- 10839039 TI - The future of palliative care nursing research in Australia. AB - Nursing research in palliative care is becoming more important as cost effectiveness and evidence-based practice are becoming mandatory. Fifty-nine published and unpublished projects relating to nursing research on palliative care in Australia between 1990 and 1996 were reviewed. Findings indicated that nurses working in palliative care in Australia are interested in researching a range of topics using varied methods. The nurses' primary interests appear to be their own professional issues and management of the patient's pain. Little research on families/carers of palliative care patients was evident. There is a need for future research to focus on projects that justify the nurse's role in the palliative care team, show that nursing interventions for symptom control affect patient outcomes and prove nurses are integral to the psychosocial and spiritual wellbeing of the patient and family during the palliative care journey. PMID- 10839040 TI - European snapshot homeless survey: results of questions asked of passers-by in 11 European cities. AB - Passers-by were interviewed outside McDonalds restaurants in 11 European cities during November and December 1998. Eight questions were asked, mainly exploring the stigma levels of a particularly socially and economically excluded group (homeless people) and also the preferred remedies for homelessness. Answers varied, especially between the countries in Western Europe and those in the former Warsaw Pact countries. Stigma levels were very high in Bucharest, Kiev and Zagreb, presumably making resettlement work difficult. People in these cities saw homeless people as 'dangerous': repositories of infectious disease and likely to make unprovoked attacks. Most passers-by also believed homeless people were under threat, especially from the police, sometimes from other passers-by, from exposure to the winters and, in Cambridge, Vienna and Zagreb, from malnutrition. Suggested remedies were increased employment, improved training and increased affordable housing, rather than the imprisonment of beggars. PMID- 10839041 TI - Perceptions of registered nurses working with assistive personnel in the United Kingdom and the United States. AB - This study examines registered nurse perceptions of their role in acute care hospitals that use nursing care assistants (NCA) and unlicensed assistive personnel (UAP). Also studied was registered nurse (RN) satisfaction with nursing care assistants and unlicensed assistive personnel in the United Kingdom (UK) and the United States of America (USA). The purpose of this study is to assist RNs and managers in the re-design of health-care delivery systems by investigating: 1. The differences and similarities of registered nurses in the UK and the USA in the perceptions of changes in the RN role when working with nursing care assistants or unlicensed assistive personnel. 2. The differences between and similarities of registered nurses in the UK and the USA in perceptions of NCA and UAP abilities to perform delegated duties, to communicate pertinent clinical information and to provide more time for professional nursing activities. Registered nurse perceptions in the UK were compared with the findings of a previous study of RN role changes and satisfaction in the USA. Registered nurses in the UK did not perceive a profound change in their role when working with UAP and were more satisfied with their use than were RNs in the USA. PMID- 10839042 TI - Transformation of story to practice: an innovative approach to long-term care. AB - The negative effects of institutionalization in a long-term care facility include depersonalization, decreased self-concept and self-esteem, and decreased psychological well being. To modify these effects, an innovative narrative approach to long-term care in the format of story telling and listening is proposed. Narrative practice, in the format of story telling and listening, leads to an ethics-based care that prioritizes the resident's personal meanings, values staff-patient relationships, and permits the resident to define quality care. The purpose of this article is to demonstrate how the functions of storytelling can be applied in the long-term care setting using the themes that emerge in the life story of a particular resident. These themes, findings of a phenomenological study, were incorporated into the resident's care plan by staff and, consequently, quality of care improved. Recognizing the value of story and incorporating the themes that emerge into a personalized care plan transforms nursing practice. PMID- 10839043 TI - Quality of life and successful aging in long-term care: perceptions of residents. AB - This study explores factors related to quality of life and successful aging in long-term care from the perspective of residents. Thirty-two residents from a long-term care facility rated factors related to quality of life and responded to open-ended items on quality of life and successful aging. Although experiencing health problems and disability, residents were optimistic about their health and quality of life. Factors identified as important to their quality of life included: interaction with family and friends, personal qualities, "room and board" items, and aspects of well-being. Notably, two factors--enjoying nature and being helpful to others--have not been widely reported in the literature but were identified here. Future research might explore other factors not traditionally associated with quality of life in long-term care. PMID- 10839044 TI - Use of the cognitive abilities screening instrument to assess elderly persons with schizophrenia in long-term care settings. AB - This article presents data evaluating the cognitive status of elderly persons with schizophrenia using the Cognitive Abilities Screening Instrument (CASI). Comparisons between the Mini-Mental State Exam (MMSE) and the CASI demonstrate greater specificity of the CASI in determining level of cognitive function. The authors argue that the CASI provides a comprehensive profile including nine cognitive domains. Therefore, for this understudied population in nursing homes, CASI is a more useful outcome measure for psychiatric nursing interventions designed to improve cognitive function in elderly residents with schizophrenia. Psychiatric nurses cannot hope to understand and improve what they cannot accurately assess. PMID- 10839045 TI - Effect of self-care ADLs on self-esteem of intact nursing home residents. AB - This 26-week repeated measures designed study was conducted on two units of a nursing home in a southern U.S. state. Two nursing approaches were compared on their effectiveness to foster self-care activities of daily living (ADLs) and improve self-esteem in dependent residents who, upon admission, were cognitively and physically intact and able to perform ADLs independently (intact residents). Twenty subjects, 10 on each unit, participated in the study. Each unit was randomly assigned to a nursing condition: a combination of the educative supportive system of care and behavior modification (Condition 1) and routine nursing care (Condition 2). Inservice training was provided to staff in Condition 1 but not in Condition 2. Following a 2-week baseline period, nursing staff encouraged subjects to do their targeted ADLs independently. When the groups were compared on self-care ADLs and self-esteem at the end of the study, subjects in Condition 1 did significantly more self-care and had significantly higher self esteem than those in Condition 2. PMID- 10839046 TI - The use of physical restraints to psychogeriatric patients in Hong Kong. AB - The use of physical restraint has been a controversial intervention in the nursing management of hospitalized elderly patients in many countries. This ethnographic study was conducted in one psychogeriatric ward in Hong Kong in order to explore what determines psychiatric nurses' decisions to use restraints on their elderly patients. By comparing the findings of three data sources, comprising semistructured interviews, observations, and clinical records, five main themes were identified with regard to the nurses decision of restraint use. They included the rationale of physical restraint, consideration of alternative measures, consideration of adverse consequences, ethical considerations, and policy and documentation of restraint use. The findings of this study demonstrate that nurses must question the established practice myths about restraint use being the best way to maintain patient safety. Most importantly, nurses need cognitive and ethical preparation to face different situations in which physical restraint may be used. PMID- 10839047 TI - Planning cognitive-behavioral management programs for long-term care. AB - Recent changes in the nature and scope of nursing home practices challenge long term care nurses to develop treatment programs that are both successful in enhancing residents' remaining quantity and quality of life, as well as cost effective in treating specific problems. The characteristics of behavioral therapies make them ideal for the open, community environment that characterizes many long-term care (LTC) settings. Under the guidelines of the Omnibus Budget Reconciliation Act of 1987, LTC facilities are expected to initiate behavioral management programs and to train staff in behavioral management practices. The purpose of this article is to discuss five key issues that should be considered in planning behavioral management programs for LTC facilities. PMID- 10839048 TI - Altruism and creative expression in a long-term older adult psychotherapy group. AB - This article describes the use of altruism and creative expression in an older adult psychotherapy group. These interventions, for which theoretic rationale and clinical examples are provided, are designed to decrease feelings of powerlessness that accompany life changes; to facilitate insight into feelings, coping, and preventative strategies for adaptation to change and loss; and to combat stereotypes, false beliefs, and myths imposed by a youth-oriented society. An integrated therapeutic model, combining psychodynamic, supportive, and cognitive/behavioral approaches, was used in the design of this group intervention for older adults. The authors, using rich clinical examples coupled with discussion of related theoretic background, provide the psychotherapist with an interesting perspective on this unique use of group psychotherapy with older adults. Such groups can provide a context within which to offer these patients the hope of relatedness, the opportunity to gain independence, and a heightened sense of integrity and possibility in one's later years. PMID- 10839049 TI - Shifting paradigms to neuropsychiatric nursing. PMID- 10839050 TI - Ethnicity, culture, and neuropsychiatry. AB - The purpose of this paper is to review the research on the relationships among ethnicity, culture, neuropsychiatric diagnosis, and treatment. Psychiatric nurses provide care to an ethnically and culturally diverse group of clients. Knowledge of ethnic and cultural differences are essential to diagnosis and treatment. Ethnic diversity affects psychiatric diagnosis. Cross-ethnic differences in genetics, diet, environmental exposure, and fetal, childhood, and adolescent development may result in varied experiences of psychiatric illness among ethnic groups. Ethnic diversity also affects psychiatric treatment. There are dramatic ethnic differences in the metabolism of psychotropic medications and the effects of drugs on target organs. These differences are again due to genetic variation, exposure to different diets and environments, and other medications in use. Cultural diversity influences both diagnosis and treatment. Cultural forces shape symptom formation and the expression of distress, creating many sources for misdiagnosis based on DSM-IV criteria. The culture-bound syndromes represent unique illness forms with a natural history distinct from DSM classification. Culture also influences treatment expectations, therapeutic compliance, family involvement, and the interpretation of side effects, all of which help determine whether or not treatment will be effective. Neuropsychiatric nurses can contribute to research by studying cross-ethnic differences and similarities in biological markers of mental illness. A second significant area for research is that of ethnicity and psychotropic drug metabolism and pharmacodynamics. PMID- 10839051 TI - Neuroreceptor function and psychopharmacologic response. AB - Psychiatric diagnoses are imprecise and that imprecision flows from current diagnostic criteria. Essentially psychiatric conditions are viewed, reported, and studied from a syndromal perspective. That is, if sufficient signs and symptoms are present, then a specific diagnosis can be made. This relative "softness" contrasts with more pathophysiology-based diagnostic criteria used by our medical surgical colleagues. As psychiatry and psychiatric nursing move toward a more scientific base for diagnosis, a greater emphasis has been placed on psychobiological dimensions of illness. The most aggressive approach to treating the psychobiological "causes" of mental disorders is the psychopharmacological approach. Essentially all psychotropic drugs affect neurotransmitter systems, however it is becoming clear that effects on receptors may be the most significant aspect of drug therapy. This article reviews receptor function and specific receptors known to be important in both the pathology and treatment of mental disorders. PMID- 10839052 TI - Neurocognitive disorders seen in HIV disease. AB - Despite dramatic improvements in the therapeutic management of clients with human immunodeficiency virus (HIV) disease, cognitive disorders still appear as clinical manifestations of the illness trajectory. The development of neuro cognitive impairment is associated with high levels of HIV activity and the resultant severe degree of immunosuppression. Although many clinicians almost exclusively associate HIV-related cognitive dysfunction with HIV encephalopathy and dementia, the etiologic conditions are numerous and include not only HIV infection affecting the brain, but also infections and neoplasms, as well as adverse effects of prescribed therapies. Treatment strategies include pharmacologic interventions, alternative and complementary therapies, and milieu management. PMID- 10839053 TI - The neurobiology of anxiety disorders: implications for psychiatric nursing practice. AB - This era of neurobiological advances challenges psychiatric nurses to develop innovative practice models that address the needs of clients with anxiety disorders. Technological discoveries and molecular and genetic research provide a wealth of evidence-based data that serve as the basis of symptom management, prevention, and health promotion and maintenance in clients experiencing anxiety disorders. This article provides an overview of current data-based studies and conceptual models of various anxiety disorders and their key components. Underpinnings include the interrelationship among neurotransmitter systems, neuroendocrinological processes, and neuroanatomical structures and their role in mediating stress, normal anxiety, and various anxiety disorders. Treatment considerations are also an integral part of this article. The discussion of pharmacologic and nonpharmacological interventions reflect dysregulation of intricate neurobiologic processes and concurrent behaviors and individual client needs. PMID- 10839054 TI - The biological basis of behavioral symptoms in dementia. AB - This article describes the pathophysiology of dementia and differentiates between cognitive and noncognitive symptoms that characterize this devastating illness. Relationships between brain anatomic and neurochemical systems and behavioral symptoms of dementia are discussed. An overview of the etiologies and neuro pathologies of dementia are presented as they relate to impairments in memory and intellectual abilities, personality changes, and behavioral symptoms. Recent genetic and molecular discoveries that have advanced our understanding of this complex spectrum of disorders and their treatment(s) are also highlighted. PMID- 10839055 TI - Perceptual reactance, drug preference, and electrodermal activity in treatment seeking substance abusers. AB - Perceptual reactance (PR) was explored as a contributing variable to drug abuse. PR is defined as the general tendency of an individual's central nervous system to perceptually modulate incoming sensory stimuli. Using an adaptive model of drug use, it was hypothesized that drug preference in substance abusers would vary with PR. Specifically, perceptual reducers, who subjectively dampen afferent sensation, would prefer experience-expanding drugs (i.e., cocaine and amphetamines) to heighten perception. Conversely, perceptual augmenters would prefer sensory-restricting drugs (i.e., opioids and sedative-hypnotics) that attenuate or modulate environmental stimuli. Utilizing subjective (Reducing Augmenting Scale) and objective (electrodermal responses) measures of PR, the relationship was tested in a sample of treatment-seeking substance abusers (n = 122). The sample was stratified on whether or not they were abstinent. Interestingly, a higher than anticipated rate of electrodermal non-responsivity was found across the sample and the implications of this finding are discussed. The study hypothesis was only supported in the electrodermally-responsive group. PMID- 10839056 TI - The family and schizophrenia. AB - There has been controversy about the role of family in the etiology and course of schizophrenia for almost 70 years. Psychoanalysts and family therapists have proposed theories about the development of schizophrenia that overtly blamed parents, and recently, expressed emotion (EE) research has been criticized as implicating families once again. However, the study of schizophrenia as a brain disorder has resulted in new understandings of the influence of the family. This article reviews recent research revealing a unique vulnerability to stress in persons with schizophrenia and suggesting that communication difficulties may be due to a shared genetic heritage. Advanced practice mental health nurses who have a solid foundation in neurobiology are ideally suited to help the person with schizophrenia and his or her family. Knowledge about the neurobiological basis of schizophrenia has become very sophisticated and complex, but that knowledge is nevertheless essential to understand the otherwise puzzling patterns of behavior shown by persons with schizophrenia. PMID- 10839057 TI - Intimate partner violence. PMID- 10839058 TI - Attachment theory and adjustment difficulties in siblings of children with cancer. AB - The demands of the childhood cancer experience on children and their parents has been investigated for a number of years. Despite this research, very little emphasis has been placed on well siblings. In the health care profession today, there is a growing perception that the psychosocial needs of the healthy siblings of children with cancer are less sufficiently met than those of other members of the family system. Previous research proposes that well siblings are especially susceptible to a number of adjustment difficulties (such as depression, anger, anxiety, feelings of guilt, and social isolation) (Murray, 1999). Given these findings, the question arises as to whether the adjustment difficulties seen in siblings are a result of the loss of, or separation from, the attachment figure- the mother who is busy caring for the child with cancer. The purpose of this article is to use attachment theory as a conceptual framework to try to understand the effects of the childhood cancer experience on siblings. Recent findings regarding siblings of children with cancer and some speculations regarding clinical implications are provided. PMID- 10839059 TI - A biobehavioral framework for examining altered sleep-wake patterns in homeless women. AB - The interactions among psychosocial and environmental stressors and the hypothalamus-pituitary-adrenal-ovarian-immune axes may provide a framework for examining altered sleep-wake patterns in women. Based on this biobehavioral framework, homeless women represent a subgroup of the female population who have many lifestyle factors that might make them vulnerable to sleep disturbances. This article presents a study designed to investigate the self-reported sleep patterns and lifestyle factors associated with the sleep of 50 homeless women, 18 44 years of age, residing in the downtown area of Los Angeles. Results indicated that almost half of the women slept six or less hours a day and had a day/night sleep pattern. A majority of the women reported restless sleep. Drinking beer, using cocaine, anxiety, depression, loneliness, and concerns about safety and money were factors significantly related to altered sleep patterns. PMID- 10839060 TI - Stress and mental health: a biobehavioral perspective. AB - The influence of stress on all aspects of health and the importance of understanding the complex interaction of the mind and body has increasingly become an issue of worldwide concern. This article offers an overview of the stress response, emphasizing the interdependence of the neurobiological components--neurologic, neuroendocrine, and endocrine axes--with emotional, behavioral, and cognitive responses. Common measurements of stress are presented, including instruments that assess stressors, cognitive/affective dimensions, biological systems, and allostatic load. Understanding the role of perceived stress and the relationship between biological and psychosocial dimensions of stress and mental disorders expands the potential for effective interventions by mental health nurses. PMID- 10839061 TI - Owls, larks and the significance of morningness/eveningness rhythm propensity in psychiatric-mental health nursing. AB - In the last decade there has been an upsurge in the research focusing on the interplay between the human circadian timing system and behavioral patterns in health and illness. Of particular interest in this area of inquiry is the overlay of what has been termed chronotype. What this refers to is the propensity of biological rhythms to express themselves in certain patterns of behavior. Commonly, these patterns have received names such as owl (evening chronotype) or lark (morning chronotype). Many people are neither a strong morning nor evening chronotype. If illness represents a change in the way a person's body functions within a given environment, then it is reasonable to believe that an "owl's" symptom presentation may vary significantly from the patterns of a "lark" who becomes ill. Recognizing that psychiatric nurses at both the generalist and the advanced practice levels have a strong interest in patterns of behavior, it stands to reason that using a lens that incorporates notions of the body's clock becomes essential. The interplay between the body's timing system and the thousands of other psychobiological rhythmic functions occurring everyday and within every human being is referred to as chronobiology. This article provides a primer for psychiatric nurses on issues of chronobiology related to morningness and eveningness rhythm propensity. PMID- 10839062 TI - Memory improvement in assisted living elders. AB - Part of the Cognitive Behavioral Model of Everyday Memory (CBMEM), an eight session cognitive enhancement program, entitled "Memories, Memories, Can We Improve Ours?" was tested with older adults living in an assisted living facility in the midwest. The aims of this quasi-experimental study were: to improve everyday memory, memory self-efficacy, and meta-memory. A total of 19 older adults (14 female, 5 male) with an average age of 83 years participated. For the pretest there were 16 individuals in the experimental group. The experimental group was post-tested one week after completing the intervention. At posttest memory self-efficacy scores significantly increased in the experimental group (M1 = 52.13, M2 = 68.50, where M1 represents pretest and M2 represents posttest). Total memory performance scores were not significantly different at posttest; however the prospective memory items of asking for an appointment (M1 = .56, M2 = 1.25), asking for a belonging (M1 = .62, M2 = .88), and delivering a message (M1 = 1.00, M2 = 1.19) significantly improved. PMID- 10839063 TI - Nurses' knowledge of pain management: implications for staff education. AB - Among the 217 nurses responding (one third of those queried), average scores on the NKAS were slightly higher (68%) than that reported in the literature but still inadequate. Nurses' greatest knowledge deficits were related to pharmacotherapeutics. Knowledge scores were uncorrelated with self-assessed level of knowledge about pain. Strategies for re-dressing these nurses' knowledge through Department of Nursing Education interventions are explored. PMID- 10839064 TI - The challenges and rewards of institutional collaboration in distance education. AB - Faculty from five nursing schools created an innovative RN to BSN program, which is offered via distance education technology. The development of a distance education academic program leading to a degree involved working to overcome many barriers; however, this strategy for delivering an academic degree has a promising future. PMID- 10839065 TI - Developing meaningful age-appropriate competencies for clinical services. AB - The process used by one large urban hospital to develop a comprehensive program for all patient care services related to the delivery of age appropriate care is discussed. The experience of this organization in identifying age appropriate competencies, educating staff about these competencies, ensuring that all staff providing direct patient care were able to demonstrate these competencies in their practices, and documenting each step of the process can serve as a guide to other organizations facing similar challenges. PMID- 10839066 TI - Fall risk assessment of hospitalized adults and follow-up study. AB - One-hundred seventy-seven randomly selected hospitalized adult patients were assessed for their risk of falls by 24 registered nurses (RNs). Twelve specific indices of fall risk assessment were studied. Patients' scores on three separate fall risk-assessment scales also were calculated. Only one of the fall risk assessment scale scores was significantly associated with documented implementation of the hospital fall prevention protocol. A follow-up study of most of these patients' medical records by seven RNs resulted in seven patients being found to have had at least one documented inpatient fall. These findings demonstrate the importance of: (1) further identifying and defining indices of fall risk assessment, (2) investing staff energy and resources in a comprehensive program of education and fall prevention, and (3) identifying intervention strategies that can be documented to prevent inpatient falls. PMID- 10839067 TI - Fairy tales of storyboarding. AB - Once upon a time, there were two creative, intelligent, staff development educators who worked in a large hospital in Dayton, Ohio, and who wanted to tell a story about how to share performance improvement initiatives with others. To narrate their story, they used a problem-solving method, a coordinating theme, bright colors, graphics, clip art, photography, data collection, and a board to display their story for "The Fundamentals of Storyboarding." A properly designed poster/storyboard can successfully attract attention, stimulate interest, and impart information. Poster presentations are effective tools for educators to share their knowledge and expertise as well as demonstrate performance improvement initiatives in their organization. Join these storytellers as they open their book and present the chapters to help create a powerful poster presentation. PMID- 10839068 TI - Building RN confidence for delegation decision-making skills in practice. AB - Delegation and coordination of patient care challenge registered nurses (RNs) practicing in capitated, managed care environments. Complicating the RN role is the fact that most RNs were not taught delegation skills while in nursing school. This study addressed the effect of a structured teaching intervention on RN delegation decision making and its enhancement of knowledge and confidence in delegation to team members. The intervention significantly increased RN delegation knowledge and overall confidence in decision making. Teaching strategies are explicated for staff development educators who are charged with providing substantive leadership approaches for staff and management nurses who have been out of nursing school and in the practice setting. The role of the RN in directing the workforce has become increasingly complex in capitated, managed care systems. PMID- 10839069 TI - Pain management in perianesthesia nursing is a human endeavor. PMID- 10839070 TI - Perioperative care of patients undergoing an epidural spinal cord stimulator for angina. AB - This article explains the use of spinal cord stimulation to control angina pain. Spinal cord stimulation for control of angina pain is a new and rapidly expanding area in which nurses are often involved. The patient selection process is outlined as is the perioperative care of patients involved in the spinal cord stimulation program. This article will be of special interest to nurses working in the areas of cardiology, day-stay surgery, and neuromodulation. PMID- 10839071 TI - Aromatherapy in perianesthesia nursing. AB - Complementary therapies are being used by nurses across America to enhance patient care. Aroma-therapy is a tool for holistic nursing that appears to enhance pain control and could prove to be a useful addition to peri-anesthesia nursing practice. This report addresses the emergence of non-conventional therapies and examines the potential role for aroma-therapy in peri-anesthesia pain management. Tables provide information about essential oils and application methods. PMID- 10839072 TI - Neurogenic pain. AB - Effective preoperative communication and an awareness of the mechanisms and treatment of neurogenic pain by nursing staff directly influence the choice of intraoperative and postoperative pain management. This article describes the case study of a young woman with neurogenic pain and highlights the importance of having a working knowledge of this type of pain, its assessment, and treatment. PMID- 10839073 TI - Pain assessment and management in the newborn infant. AB - Managing the pain of a newborn is both complex and challenging because of the subtlety of pain expression in these patients and their vulnerability. This article provides an overview of the physiology of pain in the newborn, addresses pain assessment and pain-assessment tools, explores both nonpharmacological and pharmacological approaches to pain management, and finally, lays the responsibility for pain management in the newborn squarely in the lap of the professional. PMID- 10839074 TI - The effect of paracetamol, fentanyl, and systematic assessments on children's pain after tonsillectomy and adenoidectomy. AB - Various clinical protocols are used to manage early postoperative tonsillectomy and/or adenoidectomy (T&A) pain in children. Although believed to be effective, these protocols are not evidenced-based. Therefore, a double-blind, randomized, placebo controlled (2 x 2) factorial design was used (1) to evaluate the effectiveness of 2 pain protocols used interchangeably to manage early postoperative T&A pain and (2) to investigate whether nurses' systematic pain assessments improve pain management. In the first protocol children receive a loading dose (30 to 50 mg/kg) of paracetamol (acetaminophen) Formularium der Nederlandse Apothekers (Formulary of the Dutch Royal Society for the Advancement of Pharmacy) intraoperatively, followed by regular doses (70 to 100 mg/kg/24 hours) of paracetamol. In the second protocol children receive the first protocol, plus intramuscular fentanyl citrate (1 microgram/kg) intraoperatively. Subjects were 83 healthy children between the ages of 3 and 12 years, admitted for T&A as an outpatient procedure. The child's pain was measured using observation scales (Children's Hospital of Eastern Ontario Pain Scale and Face Legs Activity Cry Consolability Scale), a visual analogue scale, and self-report measures (Faces Pain Scale and Oucher). Neither pain protocol sufficiently relieved early postoperative T&A pain, and systemic pain assessments did not improve the effectiveness of analgesics. Further research evaluating the effectiveness of pain management protocols is needed. PMID- 10839075 TI - Pain management in the elderly. AB - Assessment and management of pain are among the skills that perianesthesia nurses need to deliver safe and effective care. The principles of pain management remain fairly consistent across all age groups. However, managing the pain of elderly surgical patients presents unique challenges. Many elderly people have painful chronic conditions. Consequently, during the perioperative period, the nurse must manage both the patient's chronic and acute pain. As a group, the elderly are more vulnerable to inadequate pain assessment and treatment and are more likely to have serious side effects of analgesic therapy. Therefore, this report highlights the unique aspects of pain management in elderly patients. PMID- 10839076 TI - Preoperative strategies for managing postoperative pain at home after day surgery. AB - Patients undergoing surgery, whether major or minor, require postoperative care and pain management. Most of the postoperative care for day surgery patients continues at home. The amount of care they will need, who will care for them, and the pain management strategies needed are important issues to address before surgery. The preoperative assessment nurse provides education and practical advice about pain management at home. Then, on the day of surgery, the day surgery staff reinforces written and verbal home care instructions with the patients and their home caregivers. PMID- 10839077 TI - Perianesthesia care in Kenya: issues in managing pain. AB - A complex interplay of logistic difficulties and socioeconomic factors have created a huge gap between what is known (and taught) and what is done (and practiced) about pain and its management in Kenya. This report explores this gap between theory and practice and other issues that influence the practical management of pain in Kenya. PMID- 10839078 TI - Jordanian nurses' roles in the management of postoperative pain in the postanesthesia care unit. AB - One of the most important patient care issues for nurses is pain because of its significant impact on patients' well-being. Therefore, the main purpose of this exploratory study was to describe the role of Jordanian nurses in the management of postoperative pain in the PACU. A convenience sample of 42 postoperative patients was included in this study. Forty-two nurse-patient encounters involving 20 different nurses were observed during data collection. Findings from this study indicated that nurses in Jordan pay little attention to the area of assessment and management of postoperative pain. Cultural issues were discussed in light of the perception of pain and management of postoperative pain among Jordanian patients. Recommendations related to nursing practice, education, and research were developed to improve the quality of nursing care provided to postoperative patients in Jordan. PMID- 10839079 TI - Accountability and responsibility underpins successful pain management. AB - Does having a registered nurse as a peri-anesthesia nurse make a difference to patient outcomes? Although technicians may have the technical expertise, they lack the versatility and educational preparation of a registered nurse to provide holistic care for the patient at the most crucial periods of peri-anesthesia care. This report addresses the accountability and responsibility of the peri anesthesia nurse in pain management, highlighting the need for a knowledgeable and competent practitioner so that quality patient care is given and a positive patient outcome is achieved. PMID- 10839080 TI - Pain management: ethical issues for the perianesthesia nurse. AB - Ethical aspects of pain management in the peri-anesthesia practice setting revolve around issues of competence. Nurses must have knowledge relevant to the current scope of nursing practice, changing issues and concerns, and ethical concepts and principles. Nurses and other health care providers must engage in ongoing discussion and deliberation about ethics and culturally sensitive care. Additionally, to ensure optimal outcomes, the care provided must address those aspects that are age specific. It is important not only to develop competence as related to tasks and skills, but also to develop critical thinking and decision making. All nurses need to be able to articulate a personal philosophy. They need to understand their own values and be able to anticipate the impact that ethical choice will have on their professional practice. PMID- 10839081 TI - A position statement on pain management. American Society of PeriAnesthesia Nurses. PMID- 10839082 TI - Toward a better understanding of ASPAN: its mission for 2000 and beyond. PMID- 10839083 TI - Separation and induction behaviors in children: are parents good predictors? AB - In busy surgical outpatient settings, it is important to quickly yet thoroughly assess a child's preoperative needs to determine whether a premedication or perhaps a parent's presence is warranted to facilitate a smooth anesthetic induction. This observational study of 252 children evaluated factors related to separation and induction behaviors of children. Six percent of children had difficulty separating from their parents, and 22% were uncooperative with induction of general anesthesia via face mask. Logistic regression showed that parent prediction was the best indicator of separation behavior, whereas younger age and separation behavior were both predictive of induction behavior. Findings suggest that parents are good predictors of their child's preoperative behaviors and may, therefore, provide valuable input when perioperative nurses and anesthesiologists are making decisions about potential preoperative interventions. PMID- 10839084 TI - Same-day surgery managed care: monitoring patient and system variances. AB - The Same-Day Surgery (SDS) nursing staff at a Midwestern medical center tracked the causative factors for prolonged stays (> 2 hours) in SDS. A review of retrospective data identified that postoperative patient stays averaged between 2 and 3 hours. The top three variances for prolonged stays in SDS were pain, nausea and transportation home. Based on this data, several practice changes that were incorporated to address these variances are reported in this article. PMID- 10839085 TI - Postoperative bladder distention: measurement with bladder ultrasonography. AB - Bladder distention is a common postoperative occurrence. A process improvement project was conducted at a Midwestern Veteran Affairs Medical Center to determine whether a new method for detecting bladder distention, bladder ultrasonography, was more effective than manual palpation in the perianesthesia setting. Data were collected on 494 men over a 9-month period using bladder ultrasonography. Of those patients, 19.4% had postoperative bladder distention with greater than 400 mL of urine. This compared with 1.4% of patients who had bladder distention detected during the previous year using manual palpation. Data from the project supported the use of bladder ultrasonography as being more effective than manual palpation in the assessment of postoperative bladder distention in the PACU. PMID- 10839086 TI - Latex allergy update. AB - A recent challenge in the hospital environment is the care of patients with latex sensitivity. Since the introduction of universal precautions in the late 1980s, reports of latex sensitivity have soared. The sensitization process and development of latex allergy and its effect on the hospitalized patient are currently under much discussion. Medical personnel must stay abreast of new data and product information to provide up-to-date care for patients, as well as protection for themselves. Statistics have shown a dramatic increase in the incidence of allergy to latex-containing products. Depending on the source, the incidence is thought to range from 15% to 17% for health care workers, as opposed to 1% to 3% for the general population. Other high-risk groups report as much as a 73% incidence of latex allergies. This article explores the issue of latex allergy and its impact on patient care. Signs and symptoms, as well as the etiology of allergic reactions to latex are examined. The basics of the manufacturing and packaging processes are explored, along with elements of diagnosis and screening, with emphasis on the role of the peri-anesthesia nurse. PMID- 10839087 TI - The challenging role of the perianesthesia nurse in the office-based surgical suite. AB - Office-based surgery is increasing in popularity because of technological advances, economic benefits, and convenience to the patient and surgeon. The peri anesthesia nurse practicing in the office surgical suite assumes an increasingly complex role which may include (1) quality assurance oversight, (2) office staff education, (3) implementing and monitoring infection control measures, (4) providing medical equipment upkeep by means of scheduled maintenance, and (5) overseeing of medical supplies inventory. These responsibilities may be expected in addition to the traditional role of the nurse during the preoperative, intraoperative, and postoperative period. Recent recommendations will be reviewed regarding practice considerations applicable for the peri-anesthesia nurse designed to improve patient care and safety. PMID- 10839088 TI - Nystagmus: benign or significant? AB - Nystagmus is a broad symptom describing unusual ocular movements. The peri anesthesia nurse may identify nystagmus as a preoperative condition, as an event during recovery from general anesthesia, or as a pathological finding. Understanding the physiology of nystagmus provides a basis for nursing assessment and intervention. PMID- 10839089 TI - Tissue viability on the eve of the 21st century. PMID- 10839090 TI - A clinical evaluation of an alternating-pressure mattress replacement system in hospital and residential care settings. AB - An observational study was carried out to compare wound healing on alternating pressure mattress replacement systems (APMRS) and other surfaces in an elderly population in acute and residential care settings. Subjects were assessed for the reduction in their pressure ulcers at approximately two weeks and per day, and a visual analogue scale was used to record the patients' comfort. Seven and ten subjects were allocated to the Nimbus III APMRS (Huntleigh Healthcare Ltd) in the hospital and residential care settings respectively. There was no significant difference in the healing of the subjects' sores in the two areas either at two weeks or per day. Five people were allocated to control surfaces in the hospital setting (mainly APMRS; Pegasus Airwave, Pegasus Egerton) and ten in the residential care setting (mainly alternating-pressure overlays; AlphaXcell, Huntleigh Healthcare Ltd). There was no significant difference in the healing of subjects' sores in the two areas, either at two weeks or per day. The trial APMRS was found to be equally comfortable in either setting, and in both settings the control surfaces were not regarded as significantly different in terms of comfort. These findings, from a small sample, promote discussion about the use of pressure-relieving equipment in settings where there are older people who may be at particular risk from pressure damage and where nursing interventions are less intensive and routine. PMID- 10839091 TI - Assessment of alternating-pressure air mattresses using a time-based pressure threshold technique and continuous measurements of transcutaneous gases. AB - Alternating-pressure air mattresses (APAMs) are used increasingly for the prevention and treatment of pressure sores. Laboratory evaluation techniques have centred largely on interface pressure (IP) measurement, typically analysing discrete maximum and minimum levels, or average pressure. However, since pressure relief varies with time, a time-based analysis technique has been developed for performance assessment. IP was recorded continuously, and the duration of pressures below three thresholds (30, 20 and 10 mmHg) was calculated automatically using a computerized rig. Fifteen healthy volunteers were used to evaluate the pressure-relieving characteristics of four APAMs, including one overlay. Results indicated significant differences (P < 0.001) between products when durations below the 20 and 10 mmHg thresholds were analysed, showing that some devices were only capable of momentarily relieving pressure. Maximum contact pressures on the sacrum were significantly lower (P < 0.0001) on devices whose inflation pressure was adjusted according to subject's body mass. This technique could assist in the selection of alternating or dynamic surfaces of any description with further clinical validation. PMID- 10839092 TI - Assessment of peripheral neuropathy in the diabetic foot. AB - Peripheral neuropathy is associated with major morbidity in the diabetic population. The detection of whether peripheral neuropathy is simply present or not in patients with diabetes is insufficient. What is required is a reliable measure of both the extent and severity of neuropathy, as this enables the appropriate allocation of resources and improves the quality of care for the patients who need it most. Standardized approaches involving the use of a neurothesiometer and a 10 g monofilament can provide a reliable and objective measure of the extent and severity of neuropathy, and should therefore be adopted into protocols for best practice in the assessment of neuropathy. PMID- 10839094 TI - Care of venous leg ulcers. PMID- 10839093 TI - Pressure sore risk assessment in palliative care. AB - Pressure sore prevention in palliative care is recognized as being an essential element of holistic care, with the primary goal of promoting quality of life for patient and family. Little is known about the incidence of pressure sore development and the use of pressure sore risk assessment tools in palliative care settings. The development of a risk assessment tool specifically for palliative care patients in a 41-bedded specialist palliative care unit is described. The risk assessment tool was developed as part of a tissue viability practice development initiative. The approach adopted in the validation of the Hunters Hill Marie Curie Centre pressure sore risk assessment tool was the comparative analysis of professional judgment of experienced palliative care nurses with the numerical scores achieved during the assessment of risk on 291 patients (529 risk assessment events). This comparative analysis identified the threshold for different degrees of risk for the patient group involved: low risk, medium risk, high risk and very high risk. Further work is being undertaken to evaluate the inter-rater reliability of the new tool. A number of issues are explored in this paper in relation to pressure sore prevention in palliative care: the role of risk assessment tools, the sometimes conflicting aims of trying to ensure comfort and prevent pressure sore damage, and the uncertainties faced by palliative care nurses when they are trying to maintain quality of life for the dying. PMID- 10839095 TI - Risk assessment tools in the prevention of pressure ulcers. PMID- 10839096 TI - An update on horizontal support surfaces. PMID- 10839097 TI - [Exhaustion resulting from association with and treatment of demented patients]. PMID- 10839098 TI - [Self-preservation therapy according to Romero]. PMID- 10839099 TI - [Interprofessional collaboration in ambulatory gerontopsychiatry]. PMID- 10839100 TI - [Management of time in nursing care]. PMID- 10839101 TI - [First aid and emergency personnel--a therapeutic team?]. PMID- 10839102 TI - [Working abroad? Why not The Netherlands!]. PMID- 10839103 TI - [Effects of pharmaceutical product regulations on health- and nursing-care]. PMID- 10839104 TI - [Critical discussion on preoperative nursing visits. Report of an experience]. PMID- 10839105 TI - [Acute stroke station--stroke unit]. PMID- 10839106 TI - [Three months as "Scrub Nurse" in London]. PMID- 10839107 TI - [Nursing research studies in the USA. Experience of a scholarship student]. PMID- 10839108 TI - [Homelessness as self care problem]. PMID- 10839109 TI - [The self-image of nursing and its historical development]. PMID- 10839110 TI - [Nursing and its socio-political mission]. PMID- 10839111 TI - [Situation and significance of nursing theories and nursing philosophy]. PMID- 10839112 TI - ["I was always with body and soul a dedicated nurse"]. PMID- 10839113 TI - [Hilde Steppe--a consolation to the next generation and honour to Germany]. PMID- 10839114 TI - [In memory of Hilde Steppe]. PMID- 10839115 TI - [Nursing during national socialism. Appreciation of the activities of Prof. Dr. Hilde Steppe]. PMID- 10839116 TI - Foreign-trained Jewish nurses in Palestine and Israel 1900-1965. PMID- 10839117 TI - Preparation of methyl 2,3-di-O-mesyl-4,6-thioanhydro-alpha-D-galactopyranoside and methyl 2-O-mesyl-4,6-thioanhydro-alpha-D-gulopyranoside. AB - Two 2-oxa-7-thiabicyclo[4.2.0]octane derivatives, 4 and 10, with the D-galacto and D-gulo configuration, respectively, were obtained from methyl alpha-D glucopyranoside. The thietane cyclization involved a thio-Mitsunobu reaction resulting in a 6-thioacetate, which underwent selective base-catalyzed intramolecular nucleophilic substitution at a C-4 mesylate. The structures of 4 and 10 were elucidated by X-ray diffraction analysis. PMID- 10839118 TI - Synthesis of the tetrasaccharide related to the repeating unit of the antigen from Shigella dysenteriae type 5. AB - Starting from L-rhamnose, D-mannose and 2-amino-2-deoxy-D-glucose hydrochloride, two disaccharide blocks, namely, ethyl 2,4-di-O-benzyl-3-O-[(R)-1 (methoxycarbonyl)ethyl]-alpha-L-rhamnopyranos yl-(1-->3)-2-O-acetyl-4,6-di-O benzyl-1-thio-alpha-D-mannopyranoside and 2-(trimethylsilyl)ethyl 2-O-acetyl-3,6 di-O-benzyl-alpha-D-mannopyranosyl-(1-->3)-4,6-di-O-benzy l-2-deoxy-2-phthalimido beta-D-glucopyranoside, were synthesised and then allowed to react in the presence of N-iodosuccinimide and trifluoromethane sulfonic acid to give a tetrasaccharide derivative. This compound was converted into 2 (trimethylsilyl)ethyl 2,4-di-O-benzyl-3-O-[(R)-1-(methoxycarbonyl)ethyl]-alpha-L rhamno- pyranosyl-(1-->3)-2-O-acetyl-4,6-di-O-benzyl-alpha-D-mannopyranosyl-(1-- >4)-2-O-acetyl-3,6-di-O-benzyl-alpha-D-mannopyranosyl-(1-->3)-2-acetamid o-4,6-di O-benzyl-2-deoxy-beta-D-glucopyranoside, which on hydrogenolysis, afforded the methyl ester 2-(trimethylsilyl)ethyl glycoside of the tetrasaccharide related to the repeating unit of the O-antigen from Shigella dysenteriae type 5. PMID- 10839119 TI - Synthesis of galactofuranosyl-containing oligosaccharides corresponding to the glycosylinositolphospholipid of Trypanosoma cruzi. AB - The oligosaccharide beta-D-Galf-(1-->3)-alpha-D-Manp-(1-->2)-[beta-D-Galf- (1- >3)]-alpha-D-Manp-(1-->2)-alpha-D-Manp corresponds to the terminal end of the glycosylinositolphospholipid oligosaccharide of the protozoan Trypanosoma cruzi, the causative agent of Chagas' disease. Syntheses of methyl or ethylthio glycosides of the terminal disaccharide, trisaccharide, tetrasaccharide, and pentasaccharide corresponding to this structure are described. These syntheses employ the selective activation of a phenyl 1-selenogalactofuranoside or a phenyl 1-selenomannopyranoside donor over ethyl 1-thioglycoside acceptors with NIS-TfOH. PMID- 10839120 TI - A novel synthesis of alpha-D-Galp-(1-->3)-beta-D-Galp-1-O-(CH2)3-NH2, its linkage to activated matrices and absorption of anti-alphaGal xenoantibodies by affinity columns. AB - Pig organs transplanted into primates are rapidly rejected because of the interaction between Gal alpha(1-->3)Gal epitopes carried by the graft and natural antibodies (anti-alphaGal antibodies) present in the blood of the recipient. This report describes a simplified synthesis of the xenogeneic disaccharide and its linkage to activated gel matrices. The digalactosides alpha-D-Galp-(1-->3) alpha,beta-D-Galp-OAll were synthesized by the condensation of the trichloroacetimidoyl 2,3,4,6-tetra-O-benzyl-beta-D-galactopyranoside donor with the 3,4-unprotected allyl 2,6-di-O-benzyl-alpha- or beta-D-galactopyranoside acceptor precursor. Deallylation and hydrogenolysis led to the free digalactoside, whereas hydrogenolysis alone resulted in the 1-O-propyl digalactoside. Both products were tested by inhibition ELISA of natural anti-Gal alpha(1-->3)Gal antibodies. The alpha-D-Galp-(1-->3)-beta-D-Galp-OPr was found to be the best inhibitor. Thus, the allyl group of the partially benzylated alpha-D Galp-(1-->3)-beta-D-Galp-OAll was engineered, via the hydroxy-, the tosyloxy- and the azidopropyl intermediates, into an aminopropyl group amenable to binding to N hydroxysuccinimide-activated agarose gel matrices in order to obtain specific immunoabsorption columns. Columns made of gel substituted with 5 micromol of disaccharide per milliliter were found efficient for the immunoabsorption of anti alphaGal antibodies from human plasma. PMID- 10839121 TI - Factors determining the formation of complexes between alpha-cyclodextrin and alkylated substances in aqueous solutions: a calorimetric study at 25 degrees C. AB - The formation of complexes of alpha-cyclodextrin with cycloalkanediols, monoalkylamines and 1-alkanols has been studied calorimetrically at 25 degrees C in water, in phosphoric acid, pH 1.3, and in phosphate buffer, pH 5.5, respectively. When a complex is formed, calorimetry enables the calculation of both the enthalpy and the association constant, from which the free energy and the entropy of the process can be obtained. A model is proposed to explain the unusual trend of the association parameters for substances having alkyl chains longer than six-seven carbon atoms. The main role played by the different functional groups, and the forces involved in the association process, are discussed in the light of the signs and values of the thermodynamic parameters obtained. The effect of the variation of the aqueous medium on the hydration of the interacting substances and the consequent changes in the association parameters have been investigated. To this end, the thermodynamic parameters for the formation of the complexes between the cyclodextrin and 1-pentanol were determined at increasing concentrations of phosphate buffer. There is an increase in the association constant due to a positive entropy contribution originating from the relaxation of water molecules from the hydrophobic hydration cosphere of the alkanol to an increasingly disordered bulk. Deaquation of the interacting substances is the main factor determining the stability of the inclusion complex. PMID- 10839122 TI - The system of low-molecular-weight carrageenans and agaroids from the room temperature-extracted fraction of Kappaphycus alvarezii. AB - The room-temperature-extracted fraction from the red seaweed Kappaphycus alvarezii consists mainly of low-molecular-weight carrageenans, with structural dispersion around a basic kappa-pattern. This dispersion results from: (a) low percentages of 3,6-anhydrogalactose and the presence of precursor units; (b) important quantities of 6-O-methyl beta-D-galactose (4-sulfate) residues; (c) significant amounts of iota-repeating structure, and (d) small amounts of non sulfated and disulfated beta-D-galactose residues. Significant quantities of alpha-L-galactose units suggest the presence of agaroids, as it has been reported in several other carrageenophytes. PMID- 10839123 TI - Oligosaccharides related to xyloglucan: synthesis and X-ray crystal structure of methyl alpha-L-fucopyranosyl-(1-->2)-beta-D-galactopyranosyl-(1-->2)-alpha-D-x ylopyranoside and the synthesis of methyl alpha-L-fucopyranosyl-(1-->2)-beta-D galactopyranosyl-(1-->2)-beta-D-xy lopyranoside. AB - Trisaccharides, methyl alpha-L-fucopyranosyl-(1-->2)-beta-D-galactopyranosyl-(1- >2)-alpha-D-xy lopyranoside and methyl alpha-L-fucopyranosyl-(1-->2)-beta-D galactopyranosyl-(1-->2)-beta-D-xyl opyranoside, which are related to the side chain of xyloglucan have been synthesised. The beta-galactopyranosyl linkage of each was constructed using silver trifluoromethanesulfonate-promoted glycosylations of 2-O-acetyl-3,4,6-tri-O-benzyl-beta-D-galactopyranosyl chloride and the corresponding anomer of methyl 3,4-tri-O-benzyl-D-xylopyranoside. The resulting disaccharides were deacetylated and fucosylated using assisted halide reactions with tri-O-benzyl-alpha-L-fucopyranosyl bromide. Hydrogenolytic debenzylation of the resulting protected trisaccharides gave the methyl glycosides of the fucose-containing xyloglucan side chain. The structure of methyl alpha-L-fucopyranosyl-(1-->2)-beta-D-galactopyranosyl-(1-->2)-alpha-D-xy lopyranoside as the monohydrate was confirmed by an X-ray crystallographic study. PMID- 10839124 TI - myo-Inositol 4,6-carbonate: an easily prepared small molecule with three syn axial hydroxyl groups. AB - myo-Inositol 4,6-carbonate, a compound having three syn-axial hydroxyl groups, was synthesized in four steps suitable for gram-scale preparation. It readily forms complexes with cations. PMID- 10839125 TI - Aldol reactions on 1-deoxy-3,4:5,6-di-O-isopropylidene-L-fructose as a route to higher-carbon carbohydrates. AB - With a view to preparing higher-carbon carbohydrates, crossed-aldol reactions of the methyl ketone 1-deoxy-3,4:5,6-di-O-isopropylidene-L-fructose with a representative series of aldehydes have been investigated, and the feasibility has been demonstrated of constructing a C-11 unit containing some of the key functionality found in the carbohydrate component of the herbicidins. PMID- 10839126 TI - Monitoring hybridization during polymerase chain reaction. AB - The polymerase chain reaction (PCR) is usually analyzed by gel electrophoresis for size separation of PCR products. Additional separation techniques, such as single-stranded conformational polymorphism (SSCP), denaturing gradient gel electrophoresis (DGGE), temperature gradient gel electrophoresis (TGGE) and denaturing high-performance liquid chromatography (DHPLC), can also be used to scan for sequence alterations. These techniques are all based on the effect of PCR product hybridization on mobility. Hybridization can also be monitored with fluorescence during PCR without chromatographic or electrophoretic separation. Continuous monitoring of PCR allows the detection, quantification and sequence specificity of PCR products to be assessed, often without any need for further analysis. In such a closed system, PCR quantification with sensitivity to the single copy level can be achieved using either double-stranded DNA binding dyes or fluorescently labeled allele-specific oligonucleotide (ASO) probes. Melting curve analysis with ASO probes can be used to genotype various alleles, including single base alterations. The integration of rapid cycle PCR and ASO probes in an automated system greatly facilitates research and clinical applications of nucleic acid analysis in genetics, oncology, and infectious disease. PMID- 10839127 TI - Genetic typing and HIV-1 diagnosis by using 96 capillary array electrophoresis and ultraviolet absorption detection. AB - Current high-throughput approaches to the analysis of PCR products are based primarily on electrophoretic separation and laser-excited fluorescence detection. We show that capillary array electrophoresis can be applied to HIV-1 diagnosis and D1S80 VNTR genetic typing based simply on UV absorption detection. The additive contribution of each base pair to the total absorption signal provides adequate detection sensitivity for analyzing most PCR products. Not only is the use of specialized and potentially toxic fluorescent labels eliminated, but also the complexity and cost of the instrumentation are greatly reduced. PMID- 10839128 TI - Phytic acid as an efficient low-molecular-mass displacer for anion-exchange displacement chromatography of proteins. AB - Phytic acid, inositol-hexaphosphoric acid, molecular mass 650, a low-molecular mass compound, has been identified as a nearly ideal displacer in anion-exchange displacement chromatography for the concentration and purification of model protein mixture. The concentration of low-molecular-mass displacer is a very important parameter for successful separation by displacement chromatography. Displacer concentration influences the formation of the isotachic train and the yield and recovery of the displacement chromatographic process. There is an optimum displacer concentration in which the yield and recovery are highest. PMID- 10839129 TI - Single-cell analysis avoids sample processing bias. AB - Microscale separation tools such as capillary chromatography and capillary electrophoresis (CE) allow the study of metabolism in individual cells. In this work, we demonstrate that single-cell analysis describes metabolism more accurately than analysis of cellular extracts. We incubated HT29 cells (human colon adenocarcinoma) with a fluorescently labeled metabolic probe. This disaccharide, LacNAc, was labeled with a fluorescent dye, tetramethylrhodamine (TMR). The probe was taken up by the cells and metabolized to a number of products that retained the fluorescent label. We then split the cells into two batches. A cellular extract was prepared from one batch and analyzed by CE with laser-induced fluorescence (LIF) detection. The cells from the second batch were used for single-cell analysis by CE-LIF. Separation and detection conditions were identical for extract and single-cell analyses. We found that the electropherogram obtained by averaging the results from a number of single cells differed significantly from the cell extract electropherogram. Differences were due to sample processing during extract preparation. Disruption of the cells liberated enzymes that were compartmentalized within the cell, which allowed non metabolic reactions to proceed. The accumulation of these non-metabolic products introduced a bias in the cell extract assay. During single-cell analysis, cells were lysed inside the capillary and the separation voltage was applied immediately to separate the enzymes from their substrates and prevent non metabolic reactions. This paper is the first to report that CE analysis of single cells provides more accurate metabolic information than the CE analysis of a cellular extract. PMID- 10839130 TI - Capillary electrophoretic separation of proteins and peptides using Pluronic liquid crystals and surface-modified capillaries. AB - Separation of model mixtures of peptides/proteins carried out in a hydrophilically coated capillary in 10 mmol/l Tris and 75 mmol/l phosphate buffer containing 7.5% (w/w) Pluronic F127 copolymer (apparent pH 2.9) revealed that the separation is predominantly driven by the charge/mass ratio with little or no sieving effect. Using a coated capillary helped to remove current fluctuations that are observed in the fused-silica capillaries in the presence of the Pluronic copolymer. With peptides bearing distinct positive charge (polylysine of Mr around 3300) molecular sieving helps more detailed separation of individual species. Polyamino acids carrying negative charge can be brought to the detector window in the reversed polarity mode, however, no detailed separation of the individual species involved was observed under the conditions used. With a naturally occurring mixture of collagen fragments released by CNBr treatment of the protein the sequence of emerging peptides (positive polarity mode) with no relation to the rel. mol. mass could be revealed. It is concluded that separation of proteins/peptides in the presence of Pluronic in the background electrolyte occur on the charge/mass ratio basis with molecular sieving effects acting as a secondary partition mechanism. PMID- 10839131 TI - External electric field control of electroosmotic flow in non-coated and coated fused-silica capillaries and its application for capillary electrophoretic separations of peptides. AB - The influence of an external electric field on the electroosmotic flow in the noncoated (bare) fused-silica capillaries and in the fused-silica capillaries with covalent coating of the inner surface by the polymer of a new acrylamido derivative, N-(acryloylaminoethoxy)ethyl-beta-D-glucopyranose, has been tested in the capillary electrophoretic separations of peptide analytes. The effect of magnitude and polarity of the external electric field on the flow-rate of the electroosmotic flow, the migration times of charged analytes and the separation efficiency and resolution of separations of synthetic oligopeptides, diglycine, triglycine, glycyl-proline and prolyl-glycine, by capillary zone electrophoresis has been evaluated. The effect of the external electric field on the velocity of the electroosmotic flow was much higher in the bare fused-silica capillaries than in the coated capillaries. Better separation of the analyzed peptides was achieved in the coated fused-silica capillaries. An external electric field proved to be an effective tool for control of the electroosmotic flow and for optimization of the speed and resolution of capillary electrophoretic separations of synthetic peptides. PMID- 10839132 TI - Chromatographic assay of pharmaceutical compounds under column overloading. AB - A HPLC assay method utilizing overloaded chromatography and dual-wavelength detection was developed for a pharmaceutical formulation containing an antibacterial (clotrimazole) and a steroid (mometasone furoate) at widely different concentrations. In order to meet the limit of quantitation (LOQ) objective of not less than 0.05% of assay concentration simultaneously for both actives in the HPLC assay method, the assay concentration of the antibacterial falls into the non-linear range of its equilibrium isotherm, but still in the linear dynamic range of an ultraviolet detector. Although the analytical column is overloaded with the antibacterial and a non-symmetric elution peak is obtained, the HPLC assay method exhibits good linearity, recovery and reproducibility. PMID- 10839133 TI - Capillary electrophoretic separation of vitamins in sodium dodecyl sulfate containing buffers with lower aliphatic alcohols and n-hexane as organic modifiers. AB - The effect of lower organic alcohols as co-surfactants (methanol, ethanol, n propanol, isopropanol, propanediol, n-butanol and isoamylalcohol) and n-hexane as an organic modifier in 12.5 mol/l phosphate buffer with varying SDS concentration was investigated using a set of vitamins and p-hydroxybenzoic acid as the test mixture. It was demonstrated that optimum separations can be achieved particularly at high concentrations of the surfactant; the selectivity can be changed by adding a co-surfactant; while propanol and isopropanol show the same properties as co-surfactants, the most efficient alcohols were isoamylalcohol and propanediol. n-Butanol was capable of selective separation of p-hydroxybenzoic acid in the test mixture. Addition of ethanol appears most effective at higher concentrations (while all the other alcohols are effective already at 5% concentration, the best results with ethanol were obtained when it constituted 20% of the background electrolyte). 5% Concentration of methanol resulted in poor separation of the test mixture, however if 300 microl/10 ml of hexane were added to 20 mmol/l SDS containing phosphate buffer, the resulting separation was practically the same as with 50 mmol/l SDS. PMID- 10839134 TI - Determination of asymmetrical dimethylarginine by capillary electrophoresis-laser induced fluorescence. AB - Asymmetric dimethyl-L-arginine (ADMA) is a naturally occurring analogue of L arginine (L-Arg), the substrate of nitric oxide synthase (NOS). ADMA is a potent endogenous inhibitor of NOS and accumulates in the plasma of patients with renal failure, with peripheral arterial occlusive disease or with clinically asymptomatic hypercholesterolemia. We measured circulating concentrations of L arginine, symmetric and asymmetric dimethylarginine (SDMA and ADMA, respectively) in human serum. We developed a new method for the rapid determination of these molecules using capillary electrophoresis and laser-induced fluorescence (CE LIF). All methylated arginines were labeled with fluorescein isothiocyanate (FITC) prior to analysis. Under the capillary electrophoresis (CE) conditions used, methylated arginine derivatives were well separated, with a migration time of around 10 min. These migration times were smaller than the ones of other amino acids which do not have the same charge at pH 10. Consequently, such basic amino acids were well separated from most of the other amines or amino acids. Moreover, CE allowed one to separate all the analogues of fluorescein thiocarbamyl arginine. The results indicated that CE-LIF is useful as a selective, rapid, cheap and sensitive tool for the determination of methylated arginine products. This new technology might appreciate the endogenous substrate for NO synthase and facilitate the knowledge of the physiological and pathophysiological regulation of NO synthesis. PMID- 10839135 TI - Reaction rate, activation energy, and detection limit for the reaction of 5 furoylquinoline-3-carboxaldehyde with neurotransmitters in artificial cerebrospinal fluid. AB - The fluorogenic reagent 5-furoylquinoline-3-carboxaldehyde (FQ) has proven valuable in the analysis of proteins and small neurotransmitters. We monitored the reaction rate between this reagent and five neurotransmitters at 40 degrees and 65 degrees C in artificial cerebrospinal fluid. The reactions followed pseudo first order kinetics. The activation energy for the reaction of FQ was 10.6, 10.7, 22.0, 31.4, and 34.4 kJ mol(-1) for alanine, taurine, gamma-aminobutyric acid (GABA), glutamine, and glutamic acid, respectively. At 65 degrees C, the reaction rate was quite similar for alanine, taurine, glutamine, and glutamic acid (1.8 x 10(-3) s(-1)) but was twice as fast for GABA. A reaction time of nearly 1 h was required to quantitatively convert these neurotransmitters to their fluorescent products at 65 degrees C. Detection limits for the labeled neurotransmitters were 10(-9)-10(-8) M, which corresponded to 0.3-7 amol injected onto the capillary. PMID- 10839136 TI - Routine analysis of short-chain fatty acids for anaerobic bacteria identification using capillary electrophoresis and indirect ultraviolet detection. AB - The diagnosis of anaerobes can be difficult to perform, using classical biochemical tests. Characterization of metabolic end-products such as short-chain fatty acids (SCFA) was often used because of their reproducible biosynthesis. Despite this, SCFA are difficult to study using gas chromatography, due to their high volatility. Furthermore, the treatment of the samples are long and fastidious. Capillary electrophoresis and indirect UV detection (CE-indirect UV) is a well-known analytical method to study inorganic or organic anions. In this work, we validate the analysis of SCFA using CE-indirect UV detection. To do this, we studied the culture media of 98 anaerobic strains for the detection and quantitation of the following acids: succinic, pyruvic, acetic, lactic, propionic, 2-hydroxybutyric, butyric, 2-hydroxyvaleric, isovaleric, isocaproic, and 3-phenylpropionic. We verified that the CE-indirect UV detection analysis of SCFA for taxonomical data can be used as a mean for rapid identification for the study of anaerobes. PMID- 10839137 TI - Enantiomer separation of drugs by capillary electromigration techniques. AB - The review summarizes the most recent developments in the field of enantioseparation of chiral drugs using capillary electromigration techniques. The basic principles of enantioseparations in CE are discussed. Recent developments in sample introduction, separation and detection in capillary electrophoresis and capillary electrochromatography are summarized. The applications are arbitrarily divided into the following three groups: (a) racemates and artificial mixtures of enantiomers, (b) drug forms and (c) chiral drugs and their metabolites in biological fluids. Among the various techniques involved the relatively new developments such as CEC in aqueous and nonaqueous buffers, on-line CE-MS coupling, etc. are emphasized. PMID- 10839138 TI - Enantioselective determination of drugs in body fluids by capillary electrophoresis. AB - During the past decade, chiral capillary electrophoresis (CE) emerged as a promising, effective and economic approach for the enantioselective determination of drugs in body fluids, hair and microsomal preparations. This review discusses the principles and important aspects of CE-based chiral bioassays, provides a survey of the assays developed and presents an overview of the key achievements encountered. Applications discussed encompass the pharmacokinetics of drug enantiomers, the elucidation of the stereoselectivity of drug metabolism and bioanalysis of drug enantiomers of toxicological and forensic interest. PMID- 10839139 TI - Chiral separation of amino acids and peptides by capillary electrophoresis. AB - Chiral separation of amino acids and peptides by capillary electrophoresis (CE) is reviewed regarding the separation principles of different approaches, advantages and limitations, chiral recognition mechanisms and applications. The direct approach details various chiral selectors with an emphasis on cyclodextrins and their derivatives, antibiotics and chiral surfactants as the chiral selectors. The indirect approach deals with various chiral reagents applied for diastereomer formation and types of separation media such as micelles and polymeric pseudo-stationary phases. Many derivatization reagents used for high sensitivity detection of amino acids and peptides are also discussed and their characteristics are summarized in tables. A large number of relevant examples is presented illustrating the current status of enantiomeric and diastereomeric separation of amino acids and peptides. Strategies to enhance the selectivity and optimize separation parameters by the application of experimental designs are described. The reversal of enantiomeric elution order and the effects of organic modifiers on the selectivity are illustrated in both direct and indirect methods. Some applications of chiral amino acid and peptide analysis, in particular, regarding the determination of trace enantiomeric impurities, are given. This review selects more than 200 articles published between 1988 and 1999. PMID- 10839140 TI - Enantioselective determination by capillary electrophoresis with cyclodextrins as chiral selectors. AB - This review surveys the separation of enantiomers by capillary electrophoresis using cyclodextrins as chiral selector. Cyclodextrins or their derivatives have been widely employed for the direct chiral resolution of a wide number of enantiomers, mainly of pharmaceutical interest, selected examples are reported in the tables. For method optimisation, several parameters influencing the enantioresolution, e.g., cyclodextrin type and concentration, buffer pH and composition, presence of organic solvents or complexing additives in the buffer were considered and discussed. Finally, selected applications to real samples such as pharmaceutical formulations, biological and medical samples are also discussed. PMID- 10839141 TI - Enantiomeric separations of drugs using mixtures of charged and neutral cyclodextrins. AB - An overview on the use of mixtures of neutral and charged cyclodextrins as chiral additives for the enantioseparation of drugs by capillary electrophoresis is presented. These so called dual cyclodextrin systems can often provide unique selectivities. A brief theoretical background illustrating the influence of the chiral discrimination ability and the effective mobility of the two cyclodextrins on the overall selectivity of the enantiomeric separation is given. Typical examples of applications in the pharmaceutical field, based on the simultaneous use of a charged (cationic or anionic) and neutral cyclodextrins, are described. PMID- 10839142 TI - Separation of drug enantiomers by capillary electrophoresis in the presence of neutral cyclodextrins. AB - This is a selected review, highlighting our results obtained in an extended screening program ("The German-Chinese Drug Screening Program"), with a focus on a set of original data obtained with heptakis(2,3,6-tri-O-methyl)-beta cyclodextrin (TM-beta-CD) as the chiral solvating agent (CSA). The enantioseparation of 86 drugs by capillary zone electrophoresis in the presence of this CSA was successful for 47 drugs. The migration separation factors (alpham) and the migration retardation factors (Rm) were compared with those found for native beta-cyclodextrin (beta-CD). The patterns thus obtained were also compared with those observed for hexakis(2,3,6-tri-O-methyl)-alpha-CD (TM alpha-CD) and octakis(2,3,6-tri-O-methyl)-gamma-CD (TM-gamma-CD), respectively. From the statistical data, it can be concluded that there is a remarkable influence of the analyte structure on the electrophoretic data. A substructure 4H was found in the analyte structure that has a significant influence on the analytes' behaviour. Thus, analytes bearing the substructure 4H do not only have a strong affinity to the CDs but also a high rate of success of chiral separation in all systems reviewed. In light of this, the different ring sizes of native cyclodextrins (alpha-, beta- and gamma-CD) readily explain their behaviour towards a limited test set of chiral drugs. Sterical considerations point to the significance of side-on-binding versus inclusion in the cavity of the host. In addition to the findings from the screening program, numerous references to the literature are given. PMID- 10839143 TI - Enantiomer separation of drugs by micellar electrokinetic chromatography using chiral surfactants. AB - A review surveying enantiomer separations by micellar electrokinetic chromatography (MEKC) using chiral surfactants is described. MEKC is one of the most popular techniques in capillary electrophoresis, where neutral compounds can be analyzed as well as charged ones, and the use of chiral micelles enable one to achieve the enantioseparation. The chiral MEKC systems are briefly reviewed according to the types of chiral surfactants along with typical applications. As chiral micelles or pseudostationary phases in MEKC, various natural and synthetic chiral surfactants are used, including several low-molecular-mass surfactants and polymerized surfactants or high-molecular-mass surfactants. Cyclodextrin modified MEKC using chiral micelles is also considered. PMID- 10839144 TI - Enantiomeric separations by use of polymeric surfactant electrokinetic chromatography. AB - This review surveys the enantiomeric separation of drugs by electrokinetic chromatography using polymeric chiral surfactant pseudostationary phases. These phases have recently been shown to provide better mass transfer and increased rigidity and stability than regular micelles in micellar capillary electrophoresis. Characterization of the polymeric chiral surfactants is presented. Solution interactions of the pseudostationary phases via thermodynamics and fluorescence probe studies are evaluated. Also, case studies of enantiomeric separation of drugs using a single amino acid surfactant and the synergistic effect of the addition of gamma-cyclodextrin to the buffer is discussed. The use of dipeptide surfactants for chiral drug separations is described as well. PMID- 10839145 TI - Chiral glycosidic surfactants for enantiomeric separation in capillary electrophoresis. AB - Several glycosidic surfactants (GSs) have been shown useful in the separation of enantiomers by capillary electrophoresis. The virtue of GSs is that they can be used as (i) neutral chiral additives in the running electrolyte for the enantioseparation of charged chiral solutes by capillary zone electrophoresis, (ii) as in situ charged micelles for the enantioseparation of neutral and charged chiral solutes by micellar electrokinetic capillary chromatography (MECC), (iii) as anionic chiral surfactants in the MECC mode upon covalently attaching negatively charged groups to their sugar head groups, and (iv) as neutral and anionic chiral surfactants mixed with achiral micelles (e.g., sodium dodecyl sulfate) for MECC of enantiomers. This review article is to provide a comprehensive description of GSs in the chiral separation of various enantiomers over a wide range of operating conditions. PMID- 10839146 TI - Enantiomer separation of drugs by capillary electrophoresis using proteins as chiral selectors. AB - The separation of drug enantiomers using proteins as the chiral selectors in capillary electrophoresis (CE) is considered in this review. The proteins used include albumins such as bovine serum albumin, human serum albumin and serum albumins from other species, glycoproteins such as alpha1-acid glycoprotein, crude ovomucoid, ovoglycoprotein, avidin and riboflavin binding protein, enzymes such as fungal cellulase, cellobiohydrolase I, pepsin and lysozyme and other proteins such as casein, human serum transferrin and ovotransferrin. Protein based CE is carried out in two modes: in one proteins are immobilized or adsorbed within the capillary, or protein-immobilized silica gels are packed into the capillary (affinity capillary electrochromatography mode), and in the other proteins are dissolved in the running buffer (affinity CE mode). Furthermore, the advantages and limitations of the two modes and the factors affecting the chiral separations of various drugs by protein-based CE are discussed. PMID- 10839147 TI - Enantiomer separation of chiral pharmaceuticals by capillary electrochromatography. AB - Enantiomer separation of chiral pharmaceuticals by capillary electrochromatography (CEC) is achieved with open-tubular capillaries (o-CEC), with packed capillaries (p-CEC) or with monolithic capillaries. In o-CEC, capillaries are coated with a thin film containing cyclodextrin derivatives, cellulose, proteins, poly-terguride or molecularly imprinted polymers as chiral selectors. In p-CEC, typical chiral HPLC stationary phases such as silica-bonded cyclodextrin or cellulose derivatives, proteins, glycoproteins, macrocyclic antibiotics, quinine-derived and 'Pirkle' selectors, polyacrylamides and molecularly imprinted polymers are used as chiral selectors. Chiral monolithic stationary phases prepared by in situ polymerization into the capillary were also developed for electrochromatographic enantiomer separation. PMID- 10839148 TI - Enantiomeric separations by nonaqueous capillary electrophoresis. AB - This paper reviews the recent advances in enantioseparations by nonaqueous capillary electrophoresis (NACE) and the effect of organic solvents on mobility of enantiomers, separation selectivity and resolution. In general, the enantioseparation systems in NACE are similar to those of aqueous capillary electrophoresis (CE) except pure organic solvents are used. The influence of important parameters such as concentration and type of chiral selectors, apparent pH, ionic strength, temperature, and control of electroosmotic flow is discussed. In addition, the reported applications of NACE separations of racemates are presented. PMID- 10839149 TI - New amphiphilic aminosaccharide derivatives as chiral selectors in capillary electrophoresis. AB - Two amphiphilic aminosaccharide derivatives were investigated as chiral selector additives in capillary electrophoresis. Each substance has a glucosamine backbone carrying three hydrocarbon chains as the hydrophobic region and three carboxylic groups as the hydrophilic region, which is an artificial biologically active compound. Using each compound as a chiral selector, the optical resolution of dansylated amino acids or new quinolone antibacterial agents (NQs) was observed. Increasing the concentration of the chiral selector or the ionic strength of running solution led to successful optical resolution. In consideration of the chemical structure of each selector and the migration behavior of the enantiomers, the resolution seemed to be based on micellar electrokinetic chromatography mode. Both selectors differed in their enantioselectivity for dansylated amino acids or NQs although the chemical structures were similar. PMID- 10839150 TI - Application of ligand-exchange capillary electrophoresis to the chiral separation of alpha-hydroxy acids and beta-blockers. AB - The application of the principle of ligand-exchange capillary electrophoresis to two substance classes is described. As chiral selector N-(2-hydroxyoctyl)-L-4 hydroxyproline-copper(II) complex was used. This principle was applied to the chiral separation of alpha-hydroxy acids and drugs containing amino alcohol structure such as beta-blockers. The enantioselectivity was found to be strongly dependent on pH corresponding to the optimal conditions for complex formation for each structure class. PMID- 10839151 TI - Enantiomeric separation by capillary electrophoresis with an electroosmotic flow controlled capillary. AB - Perfect control of electroosmotic flow (EOF) was achieved by dovetailing successive multiple ionic-polymer layer (SMIL) coated capillaries. The direction and magnitude of the EOF was perfectly controllable over the pH range 2-13. Zone diffusion was not observed, even if the inner wall of the dovetailed capillary was discontinuous, or if the sample zone passed through the connected part of the capillary because the RSDs of migration time, theoretical plates, symmetry factor and S/N of the marker were almost the same when seamless capillary and dovetailed capillary were compared. The dovetailed capillary was applied to cyclodextrin modified capillary zone electrophoresis. The control of the EOF enabled us to control both the resolution and the migration order of the enantiomers. The migration time was also controllable and, therefore, the best condition between separation and migration time could be determined by controlling the EOF. Partial filling affinity electrokinetic chromatography with a protein used as a chiral selector was also studied. The migration of the pseudostationary phase was controllable by EOF, and detection of the solute at 214 nm was possible. Therefore, the EOF-controlled dovetailed capillary has great potential to expand the application of the separation technique. PMID- 10839152 TI - Separation of enantiomers by capillary electrophoresis-mass spectrometry employing a partial filling technique with a chiral crown ether. AB - Enantiomer separations were performed by capillary electrophoresis-mass spectrometry (CE-MS) with (+)-(18-crown-6)-2,3,11,12-tetracarboxylic acid (18C6H4) as a chiral selector. In order to prevent the introduction of the nonvolatile chiral, selector, 18C6H4, into the nozzle of the CE-MS interface and/or the orifice plate, a partial filling technique was employed in this study. By the partial filling technique, the contamination caused by the nonvolatile chiral selector was avoided not only during the analysis but also during the washing of capillary with the separation solution prior to the run. Several racemic compounds having a primary amino group were successfully separated. Racemic 3-aminopyrrolidine and racemic alpha-amino-epsilon-caprolactam have no strong UV absorption, but such compounds were detected with a high sensitivity by MS detection. In this paper, the effects of the length of separation zone and those of the 18C6H4 concentration were described. As the length of the separation zone was longer or as the concentration of 18C6H4 was higher, the enantiomer resolution was enhanced more and more. However, the optimization of 18C6H4 concentration was practically enough to obtain the baseline separation. PMID- 10839153 TI - Separation of enantiomers by packed capillary electrochromatography on a cellulose-based stationary phase. AB - Separation of enantiomers was performed by applying packed capillary electrochromatography (CEC). Fused-silica capillaries of different lengths with an inner diameter of 100 microm were packed with a cellulose derivative immobilized onto macroporous silica gel. Parameters such as content of modifier in the mobile phase, concentration and pH of the buffer were varied for a set of test capillaries to determine their influence on enantioselectivity. In packed CEC the highest influence on resolution of the test racemates was found by changing the acetonitrile content, while variation of the buffer concentration mostly affects the electroosmotic velocity. The performance of packed CEC and nano-LC was also compared. Packed CEC showed much better column efficiency and enantioselectivity under similar flow/electroosmotic velocity. PMID- 10839154 TI - Investigation of an enantioselective non-aqueous capillary electrochromatography system applied to the separation of chiral acids. AB - A weak anion-exchange type chiral stationary phase (CSP) based on tert. butylcarbamoylquinine as chiral selector and silica as chromatographic support was applied to non-aqueous capillary electrochromatography. The mobile phases used consisted of acetonitrile and methanol as organic solvents, and acetic acid and triethylamine were added as background electrolytes. The influence of several experimental parameters (electrolyte concentration, acetic acid-triethylamine ratio, acetonitrile-methanol ratio and temperature) was evaluated in order to obtain improved enantioselectivity and efficiency as well as short run times for the enantiomeric separation of negatively charged chiral analytes including benzyloxycarbonyl, N-(3,5-dinitrobenzyloxycarbonyl, 9-fluorenylmethoxycarbonyl, benzoyl, acetyl and N-(2,4-dinitrophenyl) derivatized amino acids and profens. Solvent composition of acetonitrile-methanol (80:20) and enhanced electrolyte concentrations up to 600 mM acetic acid at a constant acid-base ratio of 100:1 with high applied voltages of -25 kV proved to be optimum regarding short retention times and improved efficiencies. For example, the enantiomers of Fmoc Leu could be separated in less than 10 min with a resolution factor of 6.9 and about 100000 theoretical plates per meter. PMID- 10839155 TI - Non-aqueous capillary electrophoretic enantioseparation of N-derivatized amino acids using cinchona alkaloids and derivatives as chiral counter-ions. AB - A non-aqueous capillary electrophoretic method developed with quinine and tert. butyl carbamoylated quinine as chiral selectors for the enantioseparation of N protected amino acids was applied to the investigation of other quinine derivatives as chiral additives. The optimum composition of the background electrolyte was found to be 12.5 mM ammonia, 100 mM octanoic acid and 10 mM chiral selector in an ethanol-methanol (60:40, v/v) mixture. Under these conditions, a series of chiral acids, as various benzoyl, 3,5-dinitrobenzoyl and 3,5-dinitrobenzyloxycarbonyl amino acid derivatives were investigated with regards to selectand-selector relationships and enantioselectivity employing quinine, quinidine, cinchonine, cinchonidine, tert.-butyl carbamoylated quinine, tert.-butyl carbamoylated quinidine, dinitrophenyl carbamoylated quinine and cyclohexyl carbamoylated quinine as chiral selector. PMID- 10839156 TI - Enhancement of chiral recognition by formation of a sandwiched complex in capillary electrophoresis. AB - For chiral primary amino compounds not separable by cyclodextrins alone, chiral recognition was successfully achieved by the formation of a sandwiched complex of the non-chiral 18-crown-6, the chiral amine and cyclodextrin (CD) [18-crown 6+amino compound+CD]. The separation of 1-methyl-3-phenylpropylamine and 1,2,3,4 tetrahydro-1-naphthylamine racemates showed the special function of the non chiral 18-crown-6 on chiral recognition. By formation of the sandwiched complex, the chiral center of 1-methyl-3-phenylpropylamine was successfully recognized, and resolution of 1,2,3,4-tetrahydro-1-naphthylamine dramatically increased. In these studies, the mobility differences of the enantiomers were evaluated as a function of the concentration of cyclodextrins with and without the 18-crown-6, and as a function of the concentration of the 18-crown-6. In addition, the separations by this method were compared to those by the chiral 18-crown-6 reagent. PMID- 10839157 TI - Direct chiral resolution of lactic acid in food products by capillary electrophoresis. AB - Chiral resolution of native DL-lactic acid was performed by capillary electrophoresis using 2-hydroxypropyl-beta-cyclodextrin as a chiral selector. Various factors affecting chiral resolution, migration time, and peak area of lactic acid were studied. The running conditions for optimum separation of lactic acid were found to be 90 mM phosphate buffer (pH 6.0) containing 240 mM 2 hydroxypropyl-beta-cyclodextrin with an effective voltage of -30 kV at 16 degrees C, using direct detection at 200 nm. In order to enhance the sensitivity, sample injection was done under a pressure of 50 mbar for 200 s. On-line sample concentration was accomplished by sample stacking. With this system, D- and L lactic acids in food products were analyzed successfully. PMID- 10839158 TI - Enantioseparation of 3,4-dihydroxyphenylalanine and 2-hydrazino-2-methyl-3-(3,4 dihydroxyphenyl)propanoic acid by capillary electrophoresis using cyclodextrins. AB - The enantiomeric separations of 3,4-dihydroxyphenylalanine (dopa) and 2-hydrazino 2-methyl-3-(3,4-dihydroxyphenyl)propanoic acid (carbidopa) by capillary electrophoresis were studied using several native, neutral and anionic cyclodextrins as chiral additives and uncoated fused-silica capillaries. The effect of the type and concentration of the cyclodextrin added to 20 mM phosphate buffer (pH 2.5) on enantioseparation and migration times was studied. A high resolution value of 15.63 was obtained for dopa enantiomers with a buffer containing 20 mM single isomer, heptakis(2,3-diacetyl-6-sulfato)-beta cyclodextrin. The enantiomers of carbidopa were separated using 20 mM carboxymethyl-beta-cyclodextrin as a chiral resolving agent. Both methods allowed the determination of 0.1% of the D-enantiomer (second migrating) in the presence of the L-enantiomer (first migrating) of dopa and carbidopa with a good precision. These methods also gave good results in terms of precision for both peak area, migration time, linearity and accuracy. PMID- 10839159 TI - Chiral capillary electrophoretic analysis of the enantiomeric purity of a pharmaceutical compound using sulfated beta-cyclodextrin. AB - A practical chiral capillary electrophoresis method using randomly sulfated beta cyclodextrin was developed for the quantitative determination of the chiral purity of a pharmaceutical compound. A systematic method development approach was conducted by modifying selected parameters such as the concentration of the chiral selectors, buffer pH, organic modifiers, buffer concentrations and type, temperature and applied voltage. The results of the investigation permitted an improved understanding of the separation mechanism. Two facile strategies for the reversal of the enantiomer elution order are also described. The optimized method was validated in terms of variability of the chiral selector, linearity, sensitivity, accuracy, recovery, ruggedness, and precision. PMID- 10839160 TI - Fast chiral separations using sulfated beta-cyclodextrin and short-end injection in capillary electrophoresis. AB - The general applicability of sulfated beta-cyclodextrin as chiral selector and short-end injection in capillary electrophoresis (CE) as a powerful screening tool for fast and efficient chiral separation of Ormeloxifene enantiomers and racemic Ormeloxifene analogues is demonstrated. Using the short-end injection procedure, all of the 16 racemic compounds studied were successfully separated with high efficiencies and with analysis times of less than 1.2 min. Furthermore, long-end injections of eight analogues named C1-C8 afforded separations with extremely high efficiencies. A statistical evaluation of the resolution values obtained in short-end and long-end injections of compounds C1-C8 showed that the sensitivity of the CE method towards structural changes in the studied molecules is intact when the chiral analysis is performed with short-end injection compared to conventional long-end injection. PMID- 10839161 TI - Design of experiments for capillary electrophoretic enantioresolution of salbutamol using dermatan sulfate. AB - Statistical experimental design was used for the optimization and for robustness evaluation of a capillary electrophoretic method developed for the enantioresolution of salbutamol. Dermatan sulfate was used as chiral selector. The goal of the study was to obtain an efficient and fast separation. An eight run Plackett-Burman matrix was used during the optimization process for the screening of the factors and to adjust the experimental domain under study. Response surface methodology was adopted after the screening phase to obtain information about how the factors percentage of chiral selector, pH and voltage affected the considered responses resolution and analysis time. The Derringer desirability function, which makes it possible to combine results obtained for properties measured on different scales, was used to simultaneously optimize the two responses. Robustness testing was carried out using a Plackett-Burman matrix. The method was found robust as regards the response resolution while voltage and chiral selector were found to be critical factors for the robustness of analysis time response. The proposed CE method permitted the complete enantioseparation of racemic salbutamol and was applied to its chiral resolution in spiked urine samples. PMID- 10839162 TI - Optimization of the cyclodextrin-assisted capillary electrophoresis separation of the enantiomers of phenoxyacid herbicides. AB - An ethylcarbonate derivative of beta-cyclodextrin (beta-CD) with three substituents per molecule, hydroxypropyl-beta-CD and native alpha-CD have been tested as resolving agents in the capillary zone electrophoresis (CZE) separation of the four enantiomers of the herbicides mecoprop and dichlorprop. The performances of the three compounds have been quantified by means of two-levels full factorial design and the inclusion constants were calculated from CZE migration time data. Possible structure of inclusion complexes have been proposed, on the basis of molecular mechanics simulations. PMID- 10839163 TI - Calculation of stability constants for the chiral selector-enantiomer interactions from electrophoretic mobilities. AB - Interactions of chiral selectors with enantiomers or with other chiral analytes, underlying their electrophoretic separations, are particular cases of interactions of dissolved species. The interaction model that describes these reversible, fast interactions is simple. Equations describing the model are also simple and applying equations for the calculation of respective equilibrium constants to experimental data is easy using computers. Obtaining the experimental mobility data, necessary for the calculation, is the critical step on the way to stability constants quantitating the strengths of interactions of chiral selectors with enantiomers and other analytes. These data are decisive for both the accuracy and precision of the calculated constants. The meaning and applicability of the particular constant depend on the type of the constant. The common method for the determination of stability constants from electrophoretic migration data is reasonable for low and medium stability constants. For stronger complexing, characterised by stability constants of the order of 10(4) l/mol, typical of affinity chiral selectors, the method becomes unreliable. For strong complexing giving constants of the order of 10(5) or higher of or higher the method is not applicable in its commonly used form. PMID- 10839164 TI - Enantioselective binding analysis of verapamil to plasma lipoproteins by capillary electrophoresis-frontal analysis. AB - Capillary electrophoresis coupled with frontal analysis was applied to the study of enantioselective binding of verapamil (VER) to plasma lipoproteins. The drug lipoprotein mixed solution, which had been in the binding equilibrium, was hydrodynamically introduced into a non-coated fused-silica capillary. Since VER is positively charged in the neutral run buffer (pH 7.4), the unbound VER enantiomers migrated toward the cathodic end much faster than negatively charged lipoproteins and their bound forms. Once unbound VER migrated apart from lipoprotein, the bound VER was quickly released from the protein to maintain the binding equilibrium. Thus, VER migrated as a zone through the capillary and gave a trapezoidal peak with a plateau region on the electropherogram. The VER concentration in this plateau region was equal to the unbound VER concentration in the initial sample solution. It was found that the bindings of VER to high density lipoprotein (HDL), low-density lipoprotein (LDL) and oxidized LDL were not site-specific and not enantioselective. Partition-like binding to lipid part of these lipoproteins seemed to be dominant. The total binding affinities of LDL to VER were about seven-times stronger than those of HDL, and the oxidation of LDL by copper ion enhanced the binding affinities significantly. PMID- 10839165 TI - Mechanistic study of opposite migration order of dimethindene enantiomers in capillary electrophoresis in the presence of native beta-cyclodextrin and heptakis(2,3,6-tri-O-methyl)-beta-cyclodextrin. AB - The possible mechanisms of the opposite affinity pattern of the enantiomers of dimethindene [(R,S)-N,N-dimethyl-3[1(2-pyridyl)ethyl]indene-2-ethylamine] (DIM) towards native beta-cyclodextrin (beta-CD) and heptakis(2,3,6-tri-O-methyl-)-beta CD (TM-beta-CD) were studied using capillary electrophoresis (CE), NMR spectrometry, electrospray ionization mass spectrometry (ESI-MS) and X-ray crystallography. NMR spectrometry allowed to estimate the stoichiometry of the complex and to determine the binding constants. As found using ESI-MS, together with more abundant 1:1 complex, a complex with 1:2 stoichiometry may also be present in a rather small amount in a solution of DIM and beta-CD. One dimensional ROESY experiments indicated that the geometry of the complexes of DIM with native beta-CD depends on the ratio of the components in the solution. In the 1:1 solution of DIM and beta-CD the complex may be formed by inclusion of the indene moiety of DIM into the cavity of beta-CD on the primary side and into the cavity of TM-beta-CD into the secondary side. The most likely structural reason for lower affinity of the enantiomers of DIM towards the cavity of TM-beta-CD compared to native beta-CD could be elucidated. The indene moiety does not enter the cavity of TM-beta-CD as deeply as the cavity of beta-CD. This may be the most likely explanation of significantly higher affinity constants of DIM enantiomers towards the latter CD compared to the former one. The marked difference between the structure of the complexes may also be responsible for the opposite affinity pattern of the DIM enantiomers towards beta-CD and TM-beta-CD. PMID- 10839166 TI - Separation of brompheniramine enantiomers by capillary electrophoresis and study of chiral recognition mechanisms of cyclodextrins using NMR-spectroscopy, UV spectrometry, electrospray ionization mass spectrometry and X-ray crystallography. AB - Opposite migration order was observed for the enantiomers of brompheniramine [N [3-(4-bromphenyl)-3-(2-pyridyl)propyl]-N,N-dimethylamine] (BrPh) in capillary electrophoresis (CE) when native beta-cyclodextrin (beta-CD) and heptakis(2,3,6 tri-O-methyl)-beta-CD (TM-beta-CD) were used as chiral selectors. NMR spectrometry was applied in order to obtain information about the stoichiometry, binding constants and structure of the selector-selectand complexes in solution. The data were further confirmed by UV spectrometry and electrospray ionization mass spectrometry. The structure of the complexes in the solid state was determined using X-ray crystallography performed on the co-crystals precipitated from the 1:1 aqueous solution of selector and selectand. This multiple approach allowed an elucidation of the most likely structural reason for a different affinity (binding strength) of BrPh enantiomers towards beta-CD and TM-beta-CD. However, the question about a force responsible for the opposite affinity pattern of BrPh enantiomers towards these CDs could not be answered definitely. PMID- 10839167 TI - Controlling stereoselectivity with the aid of a reagent-directing group: hydroformylation, cuprate addition, and domino reaction sequences AB - The specific introduction of an appropriately designed reagent-directing group into an organic substrate allows the more efficient use of substrate direction to allow high levels of acyclic stereocontrol in both rhodium-catalyzed hydroformylation and cuprate addition to enoates. This provides access to major building blocks of the polyketide class of natural products. Incorporation of these directed reactions into sequential transformations holds promise for new particularly efficient synthetic methods. PMID- 10839168 TI - Effects of pi-bonding on stability and reactivity in AB - Recent synthetic and theoretical investigations of organometallic compounds containing the CpCr(NO) fragment have shown an interdependence of the electronic nature of ancillary ligands and the stability of a given Cr oxidation state. Understanding the correlation between ligand pi-bonding properties and the nature of the metal-based frontier orbitals permits the rationalization of observed reactivity patterns, and the identification and preparation of new classes of target molecules. PMID- 10839169 TI - Directed evolution of the fatty-acid hydroxylase P450 BM-3 into an indole hydroxylating catalyst. AB - The self-sufficient cytochrome P450 BM-3 enzyme from Bacillus megaterium catalyzes subterminal hydroxylation of saturated long-chain fatty acids and structurally related compounds. Since the primary structure of P450 BM-3 is homologous to that of mammalian P450 type II, it represents an excellent model for this family of enzymes. During studies on the directed evolution of P450 BM-3 into a medium-chain fatty-acid hydroxylase, several mutants, in particular the triple mutant Phe87Val, Leu188Gln, Ala74Gly, were observed to hydroxylate indole, producing indigo and indirubin at a catalytic efficiency of 1365 M(-1)s(-1) (kcat=2.73 s(-1) and Km=2.0 mM). Both products were unequivocally characterized by NMR and MS analysis. Wild-type P450 BM-3 is incapable to hydroxylate indole. These results demonstrate that an enzyme can be engineered to catalyze the transformation of substrates with structures widely divergent from those of its native substrate. PMID- 10839170 TI - Hydrogenation of AB - The course of the hydrogenation of [5]- and [6]metacyclophane (1b and 1c) and their thermochemistry is described. Both compounds are hydrogenated rapidly (within 10 s) to furnish the bridgehead olefins 13b and 12c. The accompanying hydrogenation enthalpies are -220 and -141 kJmol(-1), respectively. Strain energies (SE) and olefinic strains (OS) of a number of bridgehead olefins have been evaluated by DFT calculations; it was concluded that 13b belongs to the class of hyperstable olefins which correlates nicely with its reluctance to undergo hydrogenation. By combining experimental hydrogenation enthalpies and DFT calculations, SE of 187 and 121 kJmol(-1) were derived for 1b and 1c. PMID- 10839171 TI - Facilitated tip-positioning and applications of non-electrode tips in scanning electrochemical microscopy using a shear force based constant-distance mode. AB - In scanning electrochemical microscopy (SECM) a microelectrode is usually scanned over a sample without following topographic changes (constant-height mode). Therefore, deconvolution of effects from distance variations arising from non flat sample surface and electrochemical surface properties is in general not possible. Using a shear force-based constant distance mode, information about the morphology of a sample and its localized electrochemical activity can be obtained simultaneously. The setup of the SECM with integrated constant-distance mode and its application to non-flat or tilted surfaces, as well as samples with three dimensional surface structures are presented and discussed. The facilitated use of non-amperometric tips in SECM like enzyme-filled glass capillaries is demonstrated. PMID- 10839172 TI - Tautomer-dependent Bergman cyclization of novel uracil-enediyne chimeras AB - Uracil-enediyne chimeras 4, 7, and 8 were prepared and examined for their propensity to undergo Bergman cyclization. Kinetic experiments showed lactam tautomers 7 and 8 reacted up to 25 times faster than lactim ether 4. Determination of the activation energy for each cycloaromatization reaction, along with radical trapping agent dependent studies, indicate the rate differences result from different ground state energies of the starting enediynes. PMID- 10839173 TI - A new family of sequence-specific DNA-cleaving agents directed by triple-helical structures: benzopyridoindole-EDTA conjugates. AB - Sequence-specific DNA recognition can be achieved by oligonucleotides that bind to the major groove of oligopyrimidine x oligopurine sequences. These intermolecular structures could be used to modulate gene expression and to create new tools for molecular biology. Here we report the synthesis and biochemical characterization of triple helix-specific DNA cleaving reagents. It is based on the previously reported triplex-specific ligands, benzo[e]pyridoindole (BePI) and benzo[g]pyridoindole (BgPI), covalently attached to ethylenediaminotetraacetic acid (EDTA). In the presence of iron, a reducing agent and molecular oxygen, BgPI EDTA x FeII but not BePI-EDTA x FeII induced a double-stranded cut in a plasmid DNA at the single site where a triplex-forming oligonucleotide binds. At single nucleotide resolution, it was found that upon triplex formation BePI-EDTA x FeII led to cleavage of the pyrimidine strand and protection of the purine strand. BgPI-EDTA x FeII cleaved both strands with similar efficiency. The difference in cleavage efficiency between the two conjugates was rationalized by the location of the EDTA x FeII moiety with respect to the grooves of DNA (major groove: BePI EDTA x FeII, minor groove: BgPI-EDTA x FeII). This work paves the way to the development of a new class of triple helix directed DNA cleaving reagents. Such molecules will be of interest for sequence-specific DNA cleavage and for investigating triple-helical structures, such as H-DNA, which could play an important role in the control of gene expression in vivo. PMID- 10839174 TI - Enzymatic protecting group techniques for glyco- and phosphopeptide chemistry: synthesis of a glycophosphopeptide from human serum response factor. AB - The covalent modification of proteins by phosphorylation and by glycosylation with GlcNAc residues are important regulatory processes which mediate biological signal transduction. For the study of such biological phenomena in molecular detail characteristic peptides which embody both types of modification may serve as efficient tools. However, their synthesis is complicated by their pronounced acid and base lability as well as their multifunctionality. For this purpose the enzyme-labile choline ester was developed. The choline ester can be removed selectively and in high yields from various GlcNAc-glycopeptides and phosphopeptides at pH 6.5 and 37 degrees C. The conditions under which the enzymatic deprotections proceed are so mild that no undesirable side reactions are observed (i.e., no cleavage or anomerization of the glycosidic bonds and no beta-elimination of the phosphate or the carbohydrate occur). The specificity of the biocatalyst guarantees that neither the peptide bonds nor the other protecting groups present are being attacked. When this enzymatic protecting group technique was combined with the enzyme-labile 4 (phenylacetoxy)benzyloxycarbonyl (PhAcOZ) urethane protecting group a complex glycophosphopeptide could be built up. The glycopeptide is equipped with a biotin label by which it can be traced in biological systems. This peptide represents a characteristic partial structure of a glycosylated and phosphorylated sequence from the transactivation domain of serum response factor (SRF), a widely occurring human transcription factor. PMID- 10839175 TI - Interaction of 4-hydroxybiphenyl with cyclodextrins: effect of complex structure on spectroscopic and photophysical properties. AB - The structures, spectroscopic and photophysical properties of the inclusion complexes between 4-OH-biphenyl and cyclodextrins (CD) in water were studied theoretically and experimentally. The complex structures were predicted using a dynamic Monte Carlo procedure including solvation effects and analyzed via computed and experimentally determined circular dichroism. The interpretation of the induced circular dichroism spectra indicated the formation of stable 2:2 complexes between the probe and alpha-CD, while 1:1 structures account for the circular dichroism induced by beta- and gamma-CD. Fluorescence emission quantum yields and lifetimes, measured as a function of the CD concentration, confirmed the formation of this higher order complex with alpha-CD. Prototropic equilibration in the singlet excited state was found to be depressed in 2:2 complexes due to the hydrophobic environment of the OH groups, while it remained unperturbed in 1:1 complexes, where the substituent is exposed to the aqueous environment. Triplet-triplet absorption and triplet quenching data supported this interpretation. The photophysical properties of both the 1:1 and the 2:2 complexes are characterized by a significant reduction of the nonradiative decay rates. PMID- 10839176 TI - Substituent effects on two-center three-electron bonds and hydrogen bonds involving unsaturated organic functional groups and an ammonia radical cation- the resonance contribution AB - A theoretical investigation of the substituent effects on the two-center, three electron (2c-3e) bond involved between unsaturated functional groups and an amine nitrogen is presented. The competitive hydrogen-bonded complexes are also studied. In both cases, the bond energies are found to be in the range of 20-30 kcal mol(-1). The variation of these energies is discussed with respect to the electron-donating effect of the substitutents, as well as with respect to the alpha-bonded atom of the organic functional group (O, S, NH). For the 2c-3e bonds, the results are discussed on the basis of the differences of the ionization potential (delta IP) of the separated fragments and can be rationalized through the valence bond theory. For the hydrogen bonds, the substituent influence is discussed by using the differences of the proton affinities (delta PA) of the substrates. The resonating nature of the hydrogen bond in these cationic systems is investigated and is found to account for most of the binding energy. Marcus theory is compared with the proposed resonating model. PMID- 10839177 TI - Valence fluctuations and metallic behavior in K6Cu12U2S15, a new quaternary sulfide with a unique three-dimensional cubic framework AB - Mixed S2-/S1- oxidation states have been discovered in the new quaternary compound K6Cu12U2S15. Synthesized from the reaction of Cu and U in a molten alkali metal/polysulfide flux, the compound crystallizes in the cubic space group Ia3d (no. 230) with a = 18.642(7) angstroms. Its complex structure is built from [US6] octahedra connected into one-dimensional columns with [CuS3] trigonal planar units, which also serve as inter-column connection points for the extended three-dimensional structure. Magnetic susceptibility measurements reveal Curie Weiss paramagnetic behavior at temperatures above 100 K with a microeff of 2.96 microB per formula. This corresponds to 2.1 microB per U atom, and it is insufficient to distinguish between U4+ and U5+ paramagnetic ions. The conductivity of hot-pressed polycrystalline pellets is approximately 1500 Scm(-1) at room temperature, and increasing conductivity with decreasing temperature is observed, indicating metallic behavior for the material. Small and positive values for the material's thermopower (3-4 microVK(-1) from 100-300 K) also confirm p-type metallic behavior. PMID- 10839178 TI - High-frequency EPR spectra of AB - A detailed multifrequency high-field-high-frequency EPR (95-285 GHz) study has been performed on the single-molecule magnet of formula [Fe8O2(OH)12(tacn)6]Br8 x 9H2O, in which tacn = 1,4,7-triazacyclononane. Polycrystalline powder spectra have allowed the estimation of the zero-field splitting parameters up to fourth order terms. The single-crystal spectra have provided the principal directions of the magnetic anisotropy of the cluster. These results have been compared with an evaluation of the intra-cluster dipolar contribution to the magnetic anisotropy; this suggests that single-ion anisotropy is the main contributor to the magnetic anisotropy. The role of the transverse magnetic anisotropy in determining the height of the barrier for the reversal of the magnetization is also discussed. PMID- 10839179 TI - Which electron count rules are needed for four-center three-dimensional aromaticity? AB - A series of charged and neutral four-center n-electron (4c-ne, n = 1-4) molecules based on the adamantane framework, but which include combinations of boron, nitrogen, and phosphorus atoms at bridgehead positions, were studied computationally at the B3LYP/6-31G* level of density functional theory (DFT). The three-dimensional aromaticity, observed earlier for the 1,3,5,7 bisdehydroadamantane dication (1), is found to be general for 4c-2e electron systems. The degree of electron delocalization, evaluated by energetic, geometric, and various magnetic criteria, is quite independent of the molecular symmetry (point groups vary from Td to Cs), the degeneracy of the orbitals, the molecular charges, and the nature of the atoms participating in the delocalized bonding. Although the multiple positive (e.g., in 1 and some of the heteroatom systems) and multiple negative charges are strongly repulsive, the rigid adamantane frameworks help hold the bridgehead atoms within bonding distances with the fewer available electrons. The corresponding 4c-1e doublets are approximately half as aromatic as the 4c-2e singlets based on the same criteria. However, the three-electron systems may either adopt distorted but still four center delocalized structures, or alternative 3c-2e two-dimensional arrangements in which the fourth bridgehead atom is more distant. There is no need to derive special rules for each point group for 4c-ne systems. Although the three dimensional stabilization is computed to be quite appreciable, ranging between 10 and 50 kcalmol(-1), this delocalization energy is generally not sufficient to overcome distortion due to strain in higher homologues of 1 and in analogous noncage systems. Among the various 4c-2e homoadamantanedehydro dications studied, only the 1,8-dehydrohomoadamandiyl-3,6-dication forms a three-dimensional aromatic system. PMID- 10839180 TI - Charge-transfer interactions in face-to-face porphyrin-fullerene systems: solvent dependent luminescence in the infrared spectral region AB - The cyclophane-type molecular dyads 1 x 2H and 1 x Zn, in which a doubly bridged porphyrin donor adopts a close, tangential orientation relative to the surface of a fullerene acceptor, were prepared by Bingel macrocylization. The porphyrin derivatives 2 x 2H and 2 x Zn with two appended, singly linked C60 moieties were also formed as side products. NMR investigations revealed that the latter compounds strongly prefer conformations with one of the carbon spheres nesting on the porphyrin surface, thereby taking a similar orientation to that of the fullerene moiety in the doubly bridged systems. Cyclic voltammetric measurements showed that the mutual electronic effects exerted by the fullerene on the porphyrin and vice versa are only small in all four dyads, despite the close proximity of the donor and acceptor components. The steady-state and time resolved absorption and luminescence properties of 1 x Zn and 2 x Zn were investigated in toluene solution and it was shown that, upon light excitation, both the porphyrin- and the fullerene-centered excited states are deactivated to a lower-lying CT state, emitting in the IR spectral region (lambda max = 890 and 800 nm at 298 and 77 K, respectively). In the more polar solvent benzonitrile, this CT state is still detected but, owing to its very low energy (below 1.4 eV), is not luminescent and shorter-lived than in toluene. The remarkable observation of similar photophysical behavior of 1 x Zn and 2 x Zn suggests that a tight donor-acceptor distance cannot only be established in doubly bridged cyclophane type structures but also in singly bridged dyads, by taking advantage of favourable fullerene-porphyrin ground-state interactions. PMID- 10839181 TI - Macrocyclization of alpha-(alkynyloxy)silyl-alpha-diazoacetates by inter /intramolecular AB - Thermally induced intra-/intermolecular [3 + 2] cycloaddition reaction sequences of alpha-(alkynyloxy)silyl-alpha-diazoacetates 1 lead to [3.3](1,4)pyrazolophanes (2)2 and higher cyclooligomers thereof [(2)n, n > 2]. In most cases, the cyclodimer was isolated by crystallization, while a complete separation of the mixture of the higher cyclooligomers was not possible. Solid state structures of cyclodimers (2b)2 and (2c)2, cyclotrimer (2b)3, and cyclotetramer (2e)4 were determined by X-ray diffraction analysis. Field-desorption mass spectra were used to characterize the cyclooligomer mixtures. The relative amounts of the cyclooligomers depend on the substitution pattern of the diazo compound. The cyclooligomerization reactions reported herein demonstrate, for the first time, the involvement of diazo functions in macrocyclization reactions via 1,3-dipolar cycloaddition. PMID- 10839182 TI - Antibody-catalyzed removal of the p-nitrobenzyl ester protecting group: the molecular basis of broad substrate specificity. AB - Antibody catalysts for the removal of the p-nitrobenzyl ester protecting group have been generated to accommodate a broad range of substrates. Antibody 7B9, which was elicited against p-nitrobenzyl phosphonate 1, catalyzed the hydrolyses of p-nitrobenzyl monoesters of nonsubstituted, and beta- and gamma-substituted glutaric acids with almost identical Km and kcat values. In addition, 7B9 displayed substrate tolerance towards the a-substituents and accepted the p nitrobenzyl esters of Leu, Norleu, and Phe. To define the molecular basis of the broad substrate tolerance, we have cloned and sequenced the antibody and constructed a model of the active-site-hapten complex. The model showed a relatively shallow pocket of the antigen-combining site that accommodates the p nitrobenzyl moiety, and this is consistent with the observed substrate specificity. Thus, in the antibody-catalyzed reaction, the alpha-, beta-, and gamma-substituents of the substrates should be outside the combining site and ignored by the antibody recognition. A structural comparison of 7B9 with antibody D2.3, elicited against the structurally similar haptenic phosphonate, suggests the significance of the linker moiety in hapten design, which endows antibody catalysts with broad substrate specificity. These investigations provide new strategies for the generation of catalytic antibodies that accept a broad range of substrates for practical applications in organic synthetic chemistry. PMID- 10839183 TI - Pi-conjugated ligand polymers entwined around copper centres AB - We describe conjugated polymers entwined around Cu1 with alternating alpha quaterthienyl (poly[Cu(T2)2]) or 3',4',3'''',4''''-tetrahexyl-alpha-sexythienyl (poly[Cu(T3)2]) moieties and 1,10-phenanthroline complexing sites. Our strategy is to synthesise the 2,9-bis(oligothienyl)-1,10-phenanthroline precursors, then to assemble these ligands by means of Cu1 templating followed by electropolymerisation. Poly[Cu(T2)2] shows separate electroactivities for Cu redox centres and conjugated backbones, whereas the electroactivities overlap in the case of poly[Cu(T3)2]. An X-ray absorption study on these polymers in their reduced state at the Cu-K edge identifies, in both cases, four nitrogen atoms as the closest copper(I) neighbours. For poly[Cu(T2)2], the Cu1 environment is a distorted tetrahedron similar to a monomer model compound, but with a slightly higher number of steric constraints. The Cu1 environment for poly[Cu(T3)2] is a less distorted tetrahedron with an unusually short Cu1-N average bond length. Cu1 removal in poly[Cu(T2)2] induces an irreversible collapse of the structure, whereas the reversibility of Cu1 binding is almost perfect for poly[Cu(T3)2], as the hexyl chains prevent irreversible gliding of the wires after copper removal. Combined electrochemical and resistance measurements reveal that the Cu centres in poly[Cu(T2)2] play the role of mechanical support for the structure with no significant electronic interactions with the conjugated backbone, whereas in the case of poly[Cu(T3)2] copper centres contribute to the conductivity of the structure. PMID- 10839184 TI - [1]rotaxanes and pretzelanes: synthesis, chirality, and absolute configuration [In Process Citation] AB - The synthesis of aliphatically bridged [1](n)rotaxanes and (n)pretzelanes in preparative yields and the dependency of their chiroptical properties on the length (n) of their bridge are reported. A cycloenantiomeric bis(sulphonamide)[2]rotaxane with a sulphonamide group in its axle and its wheel was intramolecularly dialkylated by homologous bifunctional oligomethylene reagents to form chiral [1](n)rotaxanes bearing bridges of different lengths (n) between the axle and the wheel. Intramolecular dialkylation by 1,omega dibromoalkanes of a topologically chiral bis(sulphonamide)[2]catenane with a sulphonamide group in both of the macrolactam rings leads to pretzel shaped molecules ((n)pretzelanes) with homologous bridges between the two macrocycles. Their yields decrease with decreasing length of the bridge. The shortest bridge isolated so far in reasonable amounts consists of six methylene groups ((6)pretzelane). Remarkably, a covalent connection of axle and wheel in a [2]rotaxane was successful even with much shorter bridges-down to only three methylene groups ([1](3)rotaxane). The structural changes of the [1](n)rotaxanes with decreasing bridge length is expressed by an increasing high-field shift in the 1H NMR spectra. Enantiomeric resolution of the racemates of both series was achieved in seven cases for the [1](n)rotaxanes and two for the (n)pretzelanes by use of chiral HPLC columns. The circular dichrograms of both compound families show a strong dependency on the length of the bridge. However, the shortest bridges displayed some additional unexpected deviations. A new specification of the absolute configuration of supramolecules, such as [n]catenanes, [n]rotaxanes and (n)pretzelanes is introduced together with some nomenclature additions. PMID- 10839185 TI - Toward new materials for organic electroluminescent devices: synthesis, structures, and properties of a series of 2,5-diaryl-3,4-diphenylsiloles AB - A series of 2,5-diaryl-3,4-diphenylsiloles, with various mono-substituted phenyl groups, extended pi-conjugated groups, and heteroaryl groups as aryl groups at the 2,5-positions, has been prepared by a one-pot synthesis from bis(phenylethynyl)silanes based on the intramolecular reductive cyclization followed by the palladium-catalyzed cross-coupling with aryl halides. Crystal structures and chemical reactivities toward the alkaline desilylation reactions have been studied on the 2,5-bis(p-mono-substituted phenyl)silole derivatives to elucidate the effects of the p-substituents. The UV-visible absorption and fluorescence spectra, and cyclic voltammetry of the 2,5-diarylsiloles have been systematically evaluated. Their photophysical properties as well as their electronic structures significantly depend on the nature of the 2,5-aryl groups. PMID- 10839186 TI - Naked Au55 clusters: dramatic effect of a thiol-terminated dendrimer AB - Reaction of the thiol-terminated fourth-generation dendrimer 2-G4 (96 SH groups) with the gold cluster compound Au55(PPh3)12Cl6 in a 3:1 molar ratio in dichloromethane results in the formation of bare Au55 clusters. The cuboctahedrally shaped Au55 particles coalesce to well-formed microcrystals (Au55) infinity. The role of the dendrimer is not only to remove the phosphine and chlorine ligands but also to act as an ideal matrix for perfect crystal growth. Transmission electron microscopy (TEM), small- and wide-angle X-ray diffraction (SAXRD and WAXRD) measurements indicate a structure where rows of edge-linked Au55 building blocks form a distorted cubic lattice. The X-ray data fit best if a 5% reduction of the Au-Au bond length in the Au55 clusters is assumed, in agreement with previous extended X-ray absorption fine structure (EXAFS) measurements. Energy-dispersive X-ray spectroscopy (EDX) analyses and IR investigations show the absence of PPh3 and Cl in the microcrystals. PMID- 10839187 TI - Redox states and associated interchain processes of thienylenevinylene oligomers AB - The electronic structure of successive redox states of two series of thienylenevinylene oligomers (nTVs, n=4, 5, 6, 8, and 12) that carry hexyl substituents at the alpha- or beta-positions in order to increase the solubility was investigated in detail by means of UV/ Vis/near-IR and ESR spectroscopy in solution. The nTV redox states have been fully characterized up to the dication for the shorter oligomers (n < or = 6) and up to the tetracation for the longer oligomers (n > or = 8). While the monocation radicals of the nTVs exhibit two dipole-allowed electronic transitions in the Vis/near-IR region, all the higher oxidized states invariably show a single strong absorption in the near-IR region. The electronic transitions of the various oxidized states and those of their vibronic replicas shift to lower energies with increasing conjugation length. The ESR spectra, recorded as a function of the degree of oxidation, provide evidence for the presence of electron spin in the odd-charged and the absence of spin in even-charged cations. Variable-temperature UV/Vis/near-IR and ESR spectroscopy establish that the tendency of the nTV monocation radicals to form spin-less alpha dimers in solution strongly depends on the number of solubilizing hexyl groups. While the oligomers that carry two hexyl chains at the a-positions of the terminal thiophenes (alpha-nTVs) readily form pi dimers at low temperature, the oligomers that carry hexyl groups on the beta- and beta'-positions of every thiophene ring (beta-nTVs) do not form pi dimers. Low-temperature UV/ Vis/near-IR and ESR experiments on solutions in which neutral and singly oxidized nTVs are simultaneously present, reveal the occurrence of interchain interactions between these two species, accompanied by a pronounced change in the existing disproportionation equilibrium. PMID- 10839188 TI - Stimulatory effect of regucalcin on proteolytic activity is impaired in the kidney cortex cytosol of rats with saline ingestion. AB - The effect of regucalcin (RC) on neutral proteolytic activity in the cytosol of rat kidney cortex was investigated. Proteolytic activity was significantly increased by the presence of RC (0.01-0.10 microM) in the enzyme reaction mixture. This increase was completely abolished by the addition of anti-RC monoclonal antibody (150 ng/ml). When the renal cortex cytosol was incubated without RC addition, the degradation of globin of substrate was demonstrated by sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis. This degradation was clearly inhibited by the addition of anti-RC antibody (150 ng/ml), indicating that protein degradation results partly from the cytosolic endogenous RC. Meanwhile, proteolytic activity was significantly decreased in the renal cortex cytosol of rats with saline ingestion for 2, 7, and 14 days. The effect of RC (0.1 microM) in increasing proteolytic activity was weakened in the kidney cortex cytosol of saline-ingested rats. The present study suggests that endogenous RC plays a role in the activation of proteases in the renal cortex cytosol, and that the RC effect is impaired in saline-ingested rats. PMID- 10839189 TI - Insulin resistance in fat cells from obese Zucker rats--evidence for an impaired activation and translocation of protein kinase B and glucose transporter 4. AB - The effect of insulin on glucose transport, glucose transporter 4 (Glut4) translocation, and intracellular signaling were measured in fat cells from lean and obese Zucker rats of different ages. Insulin-stimulated glucose transport was markedly reduced in adipocytes from old and obese animals. The protein content of Glut4 and insulin receptor substrates (IRS) 1 and 2 were also reduced while other proteins, including the p85 subunit of PI3-kinase, Shc and the MAP kinases (ERK1 and 2) were essentially unchanged. There was a marked impairment in the insulin stimulated tyrosine phosphorylation of IRS-1 and 2 as well as activation of PI3 kinase and PKB in cells from old and obese animals. Furthermore, insulin stimulated translocation of both Glut4 and PKB to the plasma membrane was virtually abolished. The phosphotyrosine phosphatase inhibitor, vanadate, increased the insulin-stimulated upstream signaling including PI3-kinase and PKB activities as well as rate of glucose transport. Thus, the insulin resistance in cells from old and obese Zucker rats can be accounted for by an impaired translocation process, due to signaling defects leading to a reduced activation of PI3-kinase and PKB, as well as an attenuated Glut4 protein content. PMID- 10839190 TI - Antioxidant associated chemoprevention by selenomethionine in murine tumor model. AB - Effectiveness of selenium in different forms like sodium selenite, selenocysteine and selenomethionine has been compared in four different doses, namely 4, 6, 8 and 10 ppm of each, in terms of their bioavailability and prolongation of survival of Dalton's lymphoma (DL) bearing mice. Selenomethionine, at a dose of 8 ppm, was found to be the most bioavailable and least-cytotoxic form that was capable of increasing the life span of the tumour bearing hosts maximally (almost two-fold). Beneficiality of selenomethionine has also been studied by observing continuous changes brought about by this compound on the glutathione (GSH) level, glutathione peroxidase (GPx) activity and extent of lipid peroxidation in the hepatic tissue of the tumour bearing hosts, which are indispensable for a cell to function normally and are found to exhibit significantly altered behaviour in neoplastic cells. Selenomethionine caused the maintenance of high steady state GSH level and a normal GPx activity during the fist phase of tumour growth. It also controlled lipid peroxidation during the first 15-20 days following tumour transplantation. These conditions helped in the maintenance of intracellular redox balance, cellular integrity and metabolic rhythms of cells in DL bearing mice receiving selenomethionine. PMID- 10839191 TI - Molecular and kinetic properties of sperm specific LDH after radiation inactivation. AB - Radiation inactivation of sperm specific lactate dehydrogenase-C4 (LDH-C4) has been studied and compared with the somatic LDH in aqueous solution. D37 of C isozyme was 470 Gy and that of B isozyme was 520 Gy. Semi-log plots of log N/No versus dose suggested that the inactivation of two LDH isozymes in presence of normal saline follows a single hit kinetics. Target molecular weight calculated by radiation analysis was found as 1.52 x 10(5) gm/mole for LDH-C4 and 1.38 x 10(5) gm/mole for LDH-B4. SDS-PAGE of irradiated enzymes showed a band of 35 kDa but did not indicate the presence of any other extra band, when compared with sham-irradiated enzymes. Chemical kinetics of residual activity following irradiation at D37 showed decrease in Vmax with coenzymes and primary substrates. However, decrease in Km was seen with pyruvate as increasing substrate. Nevertheless, K did not change when NAD+ was the leading substrate for LDH-B4 or LDH-C4. A hyperchromicity in intrinsic fluorescence and a blue shift in lambdamax over sham-irradiated LDH-C4 revealed the exposure of buried tryptophan residues to the surface after radiation inactivation. Results suggest that inspite of presence of variant amino acids, the conformations of two isozymes are stabilized by similar forces which behave in a similar way for radiation inactivation in aqueous phase. PMID- 10839192 TI - Termination of asynchronous contractile activity in rat atrial myocytes by n-3 polyunsaturated fatty acids. AB - A protective effect of the n-3 polyunsaturated fatty acids (PUFAs) in preventing ventricular fibrillation in experimental animals and cultured cardiomyocytes has been demonstrated in a number of studies. In this study, a possible role for the n-3 PUFAs in the treatment of atrial fibrillation (AF) was investigated at the cellular level using atrial myocytes isolated from young adult rats as the experimental model. Electrically-stimulated, synchronously-contracting myocytes were induced to contract asynchronously by the addition of 10 microM isoproterenol. Asynchronous contractile activity was reduced following acute addition of the n-3 PUFAs docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) at 10 microM, compared with no fatty acid addition (from 99.0+/-1.0% to 30.7+/-5.2% (p < 0.05) for DHA and 23.8+/-2.8% (p < 0.01) for EPA), while the saturated fatty acid, docosanoic acid (DA) and the methyl ester of DHA (DHA m.e.) did not exert a significant effect on asynchronous contractile activity. Asynchronous contractile activity was also reduced to 1.7+/-1.7% in the presence of the membrane fluidising agent, benzyl alcohol (p < 0.001 vs no fatty acid addition). Cell membrane fluidity was determined by steady state fluorescence anisotropy using the fluorescent probe, TMAP-DPH. Addition of DHA, EPA or benzyl alcohol significantly increased sarcolemmal membrane fluidity (decreased anisotropy, r(ss)) of atrial myocytes compared with no addition of fatty acid (control) (from r(ss) = 0.203+/-0.004 to 0.159+/-0.004 (p < 0.01) for DHA, 0.166+/-0.001 (p < 0.01) for EPA and 0.186+/-0.003 (p < 0.05) for benzyl alcohol, while DA and DHA m.e. were without effect. It is concluded that the n-3 PUFAs exert anti-asynchronous effects in rat atrial myocytes by a mechanism which may involve changes in membrane fluidity. PMID- 10839193 TI - Differential responses of Bcl-2 family genes to etoposide in chronic myeloid leukemia K562 cells. AB - Etoposide is a potent anticancer agent that is used to treat various tumors. We have investigated the dose-dependent effect of etoposide on apoptosis using chronic myeloid leukemia K562 cells treated with low (5 microM) or high (100 microM) concentrations of the drug. At a low concentration, etoposide induced little apoptosis at 24 h, while about 20% of the cells showed apoptosis morphologically at a high concentration. Processing of caspase-3 was slightly detected from 12 h and became obvious at 24 h with 100 microM etoposide. Caspase 3-like protease activity was detected at 24 h with a high concentration. Moreover, these changes were accompanied by cleavage of poly ADP ribose polymerase (PARP). Changes of the mRNA levels of most apoptosis-regulating genes were not prominent at both concentrations, except for the rapid induction of c IAP-2/HIAP-1 and the down-regulation of Bcl-X(L) by 100 microM etoposide. The downregulation of Bcl-X(L) protein occurred from 6 h, while Bax protein conversely showed a slight increase from 6 h. Taken together, the present findings show that the dose-dependent apoptotic effect of etoposide is based on a change in the balance between Bcl-X(L) and Bax, which precedes the activation of caspase-3. PMID- 10839194 TI - Urate attenuates oxidation of native low-density lipoprotein by hypochlorite and the subsequent lipoprotein-induced respiratory burst activities of polymorphonuclear leukocytes. AB - Oxidation converts native low-density lipoprotein (LDL) into a signal molecule promoting inflammatory processes during atherogenesis. The exact contribution of different antioxidants in prevention of LDL oxidation is not known. Uric acid efficiently scavenges oxidants including hypochlorite. We investigated the effect of different urate concentrations (25-500 micromol/l) on the oxidation of isolated native LDL by sodium hypochlorite (1000 micromol/l). While relative electrophoretic mobility declined continuously with increasing urate concentrations in the oxidation medium, lipid peroxidation as measured by TBARS was blunted only at high molar urate/NaOCl ratios. By decreasing oxidative modifications, urate dose-dependently (beginning with a urate/NaOCl ratio of 1:40) diminished stimulatory effects of oxidized LDL on the respiratory burst of resting polymorphonuclear leukocytes (PMNL). Protecting effects of urate against the proinflammatory action of oxidized LDL on activated cells were evident only at a molar urate/NaOCl ratio of 1:2 suggesting different sensitivities of PMNL to LDL oxidation state in dependence on their activity state. PMID- 10839195 TI - Implication of tissue transglutaminase and desmoplakin in cell adhesion mechanism in human epidermis. AB - The distribution patterns of both tissue and keratinocyte transglutaminases (TGase), as well as that of desmoplakin (DP), have been immunohistochemically investigated in human skin cultured in the absence or presence of cystamine and enalapril, two acantholytic agents. In the control samples, tissue TGase is predominantly expressed in lower layers of the epidermis and is located intercellularly. Conversely, in tissues cultured with cystamine or enalapril, a diffuse cytoplasmatic staining was observed. Similarly, DP, detected on the cell membrane in the control, shifts into the cytosol of the keratinocytes following treatment. The distribution pattern of the keratinocyte enzyme in the acantholytic epidermis was identical to that observed in the normal one. Since cystamine and enalapril are TGase inhibitors and DP was shown to act as a TGase substrate in vitro, we suggest that DP and tissue enzyme may participate in cell adhesion at the intraepidermal level. PMID- 10839196 TI - Selective down-regulation of the G(q)alpha/G11alpha G-protein family in tumour necrosis factor-alpha induced cell death. AB - Investigations into the regulation of heterotrimeric GTP-binding protein alpha subunits in models of tumour necrosis factor-alpha (TNF)-induced cell death, revealed the selective down-regulation of the G(q)alpha/G11alpha family of G proteins. The human HeLa and murine L929 cells treated with recombinant human TNF for up to 24 h displayed down-regulated G(q)alpha/G11alpha family protein levels, but not G(s)alpha, G(i)alpha and G(o)alpha protein levels as determined by Western analyses. This effect of TNF was observed in a concentration--and time dependent manner, consistent with the profiles of TNF-induced cell death observed. Moreover, the functioning of G(q)alpha/G11alpha family proteins were found to be impaired in TNF-treated cells, as measured by agonist-induced [Ca2+]i release. In contrast, G(s)alpha activity was unaltered by TNF-treatment, determined by measurement of agonist-induced intracellular cyclic AMP generation. These findings in TNF-induced cytotoxic models, indicate a novel 'cross-talk' mechanism by which TNF alters Ca2+-signalling mechanisms, which may contribute towards the apoptotic and necrotic cell death. PMID- 10839198 TI - Purification, characterization and thermostability of lipase from a thermophilic Bacillus sp. J33. AB - A thermostable lipase produced by a thermophilic Bacillus sp. J33 was purified to 175-fold with 15.6% recovery by ammonium sulphate and Phenyl Sepharose column chromatography. The enzyme is a monomeric protein having molecular weight of 45 kDa. It hydrolyzes triolein at all positions. The fatty acid specificity of lipase is broad with little preference for C12 and C4. The Km and Vmax for lipase with pNP-laurate as substrate was calculated to be 2.5 mM and 0.4 microM min(-1) ml(-1) respectively. The immobilized enzyme was stable for 12 h at 60 degrees C. Polyhydric alcohols such as ethylene glycol (2.5 M), sorbitol (2.5 M) and glycerol (2.5 M) were used as thermostabilizers. Lipase acquired a remarkable stability, since no deactivation occurred at 70 degrees C for 150 min in the presence of additives. PMID- 10839197 TI - Differential agonist-induced desensitization of P2Y2 nucleotide receptors by ATP and UTP. AB - The equal potency and efficacy of the agonists, ATP and UTP, pharmacologically distinguish the P2Y2 receptor from other nucleotide receptors. Investigation of the desensitization of the P2Y2 receptors is complicated by the simultaneous expression of different P2 nucleotide receptor subtypes. The co-expression of multiple P2 receptor subtypes in mammalian cells may have led to contradictory reports on the efficacy of the natural agonists of the P2Y2 receptor to induce desensitization. We decided to investigate the desensitization of human and murine isoforms of the P2Y2 receptor, and to rigorously examine their signaling and desensitization properties. For these purposes, we used 1321N1 astrocytoma cells stably transfected with the human or murine P2Y2 receptor cDNA, as well as human A431 cells that endogenously express the receptor. The mobilization of intracellular calcium by extracellular nucleotides was used as a functional assay for the P2Y2 receptors. While ATP and UTP activated the murine and human P2Y2 receptors with similar potencies (EC50 values were 1.5-5.8 microM), ATP was approximately 10-fold less potent (IC50 = 9.1-21.2 microM) than UTP (IC50 = 0.7 2.9 microM) inducing homologous receptor desensitization in the cell systems examined. Individual cell analyses of the rate and dose dependency of agonist induced desensitization demonstrated that the murine receptor was slightly more resistant to desensitization than its human counterpart. To our knowledge, this is the first individual cell study that has compared the cellular heterogeneity of the desensitized states of recombinant and endogenously expressed receptors. This comparison demonstrated that the recombinant system conserved the cellular regulatory elements needed to attenuate receptor signaling by desensitization. PMID- 10839199 TI - Pro and antioxidant responses to repeated administration of diazepam in rat brain. AB - The role of pro/antioxidative processes during a low, subtoxic dose schedule of diazepam (3 mg/kg/day i.p.) for 7 days and its withdrawal in subcellular preparations of rat brain regions was studied in detail. The results indicated heterogeneity in the regional responses as well as in subcellular compartments. After 7 days of exposure to the drug, a decrease in the Mn-SOD activity was observed in the 3 regions studied while a significant increase in Cu/Zn-SOD activity was seen in cerebellum (CBL) and brain stem (BS) along with that of mitochondrial glutathione reductase. The post-mitochondrial fraction (PMF) showed a significant increase in GR activity in cerebrum. Enhancement of total and free thiol levels was observed in cerebrum and cerebellum whereas in BS free thiols were not enhanced. It was interesting to note that in the animals withdrawn from the drug and sacrificed after an interval of 7 days, the level of TBARS showed a highly significant increase in mitochondria of CB and CBL and 89% increase in BS. Similar trend was observed in the post-mitochondrial fractions of all the 3 regions whereas the activity of isozymes of SOD decreased (p < 0.001) in CBL and BS and to a lesser extent in CB. The GR activity was significantly decreased only in the mitochondria of cerebrum with a 34% rise in cerebellum and no change in BS. The PMFs showed a decrease in CB and CBL but a 20% rise in BS. Thus, the data show modulation of antioxidant responses during short-term administration of diazepam, and a lowering of peroxidative decomposition of polyunsaturated fatty acids of membranes. However, after withdrawal of the drug, PUFAs were found to be more vulnerable to peroxidative decomposition and changes in the antioxidant defenses were also observed, which did not come back to normal level during the study. PMID- 10839200 TI - Secretion of MCP-1 and IL-6 by cytokine stimulated production of reactive oxygen species in endothelial cells. AB - Endothelial cells are known to produce reactive oxygen species by several mechanisms. Functional consequences of increased production of reactive oxygen species were investigated in vitro after stimulation with several proinflammatory cytokines. Time dependent increases in DCF-fluorescence as a measure of reactive oxygen load were quantified in single cells after incubation with TNF-alpha, IL-1 and IFN-gamma. The increased DCF-fluorescence was inhibited by cell permeant antioxidative substances Tiron and Tempol. NMMA, an inhibitor of nitric oxide synthase reduced endothelial DCF-fluorescence only marginally, indicating a minor participation of nitric oxide production in this detection system. Cytokine induced endothelial DCF-fluorescence increased in the presence of NADH, whereas coincubation with NADPH or xanthine was without effect. Flavoenzyme inhibitor diphenyliodonium abolished stimulated DCF-fluorescence. Cytokine induced release of MCP-1 and IL-6 by endothelial cells was completely inhibited in the presence of Tiron and Tempol, whereas NMMA was less effective. Collectively these data indicate that cytokine stimulated endothelial cells increase their reactive oxygen species production probably via NADH oxidase and this production may critically be involved in the secretion of MCP-1 and IL-6. PMID- 10839201 TI - Contribution of de novo protein synthesis to the hypertrophic effect of IGF-1 but not of thyroid hormones in adult ventricular cardiomyocytes. AB - BACKGROUND: Enhanced expression of IGF-1 occurs in left ventricular hypertrophy (LVH) associated with systemic hypertension. Cardiac dysfunction accompanied by LVH is also observed in hyperthyroidism. OBJECTIVE: to assess the relative contributions of de novo protein synthesis and attenuated protein degradation to increased protein mass associated with cardiomyocyte hypertrophy elicited by IGF 1 and thyroid hormones (tri-iodo thyronine T3, and l-thyroxine T4), respectively. METHODS: total mass of protein, and both the incorporation, and removal of previously incorporated l-U-14C-phenylalanine, indices of protein synthesis and degradation, respectively, were assessed in quiescent adult rat ventricular cardiomyocytes maintained in short-term culture, and corrected for DNA content, as a index of cell number. RESULTS: IGF-1 (1 pM-100 nM) increased cell protein significantly, maximally at 1 nM and by 38% above basal value after 24 h. T3 (10 pM-2 microM) and T4 (10 pM-2 microM) increased cell protein significantly maximally at 1 microM and by 33.2 and 30.5%, respectively, above basal value. IGF 1 (< or = 10 pM), T3 (10 pM-2 microM) and T4 (10 pM-2 microM) did not increase incorporation of l-U-14C-phenylalanine above basal values. IGF-1 (100 pM-100 nM) increased incorporation of radiolabel significantly maximally at 100 nM and by 56%. T4 (100 pM) and IGF-1 (10 pM), concentrations that did not stimulate de novo protein synthesis, attenuated the degradation of radiolabelled protein by 13.6 and 11.8%, respectively, compared to control values after 48 h. CONCLUSION: These data indicate that the acute hypertrophic response to (i) thyroid hormones cannot be attributed to initiation of de novo protein synthesis; (ii) IGF- 1 comprises two components; the response elicited by IGF-1 (< 10 pM) is independent of, while the response elicited by IGF-1 (> 100 pM) is due to de novo protein synthesis. PMID- 10839202 TI - Antioxidant properties of (-)-epicatechin-3-gallate and its inhibition of Cr(VI) induced DNA damage and Cr(IV)- or TPA-stimulated NF-kappaB activation. AB - Electron spin resonance (ESR) spin trapping was utilized to investigate the scavenging effects on hydroxyl radicals (*OH) and superoxide radicals (O2*-) by ( )-epigallocatechin-3-gallate (EGCG), one of the major anticancer compounds in tea. The spin trap used was 5,5-dimethyl-pyrroline N-oxide (DMPO). The Fenton reaction (Fe2+ + H2O2-->Fe3+ + *OH + OH-) was used as a source of *OH radicals. EGCG efficiently scavenges *OH radicals with reaction rate of 4.62 x 10(11) M( 1)sec(-1), which is an order of magnitude higher than several well recognized antioxidants, such as ascorbate, glutathione and cysteine. It also scavenges O2*- radicals as demonstrated by using xanthine and xanthine oxidase system as a source of O2*- radicals. Through its antioxidant properties, EGCG exhibited a protective effect against DNA damage induced by Cr(VI). EGCG also inhibited activation of nuclear transcription factor NF-kappaB induced by Cr(IV) and 12-o tetradecanoylphorbol-13-acetate (TPA). The present studies provide a mechanistic basis for the reported anticarcinogenic properties of EGCG and related tea products. PMID- 10839203 TI - An accumulation of IGF-I and IGF-binding proteins in human umbilical cord. AB - It is known that extracellular matrix components (ECM) may serve as a storage site to concentrate and stabilize growth factors in the vicinity of cells. IGF-I is expressed in most fetal tissues and it is involved in anabolic effects on protein and sulphated glycosaminoglycans biosynthesis, cell proliferation and differentiation. We demonstrated that human umbilical cord (UC) tissues contain large amounts of IGF-I and IGF-I-binding proteins (BP-3 and BP-1). Particularly Wharton's jelly appears to be an abundant reservoir of IGF-I and BPs. Relatively low amount of cells and large amounts of collagen and glycosaminoglycans in UC tissues (especially in Wharton's jelly) suggest that IGF-I may play a major role in stimulation of these cells to produce ECM components. The specific BPs in these tissues may be important modulators of IGF-I action during fetal development. PMID- 10839204 TI - Protective effect of secoisolariciresinol diglucoside against streptozotocin induced diabetes and its mechanism. AB - OBJECTIVES: Reactive oxygen species (ROS) have been implicated in the development of streptozotocin (STZ)-induced diabetes mellitus. Secoisolariciresinol diglucoside (SDG) isolated from flaxseed is an antioxidant. An investigation was made of the effects of SDG on the development of STZ-induced diabetes in rat, to determine if SDG can prevent/reduce the development of diabetes and if this prevention/reduction is associated with reduction in oxidative stress. DESIGN AND METHODS: The rats were divided into 4 groups: Group I, Control; Group II, SDG (22 mg/kg body wt, orally) for 24 days; Group III, STZ (80 mg/kg intraperitoneally); Group IV, SDG in the dose similar to Group II three days prior to STZ and 21 days thereafter. Oxidative stress was assessed by measuring serum and pancreatic lipid peroxidation product malondialdehyde (MDA), pancreatic antioxidant reserve (pancreatic-CL) and oxygen free radical producing activity of white blood cells (WBC-CL). A diagnosis of diabetes was made on the basis of glucosuria and was confirmed at the time of sacrifice (21 days after STZ treatment) by the presence of hyperglycemia. At the end of the protocol blood samples were collected for estimation of glucose, MDA and WBC-CL, and pancreas were removed for estimation of MDA and antioxidant reserve. RESULTS: Incidence of diabetes was 100% in Group III and 25% in Group IV. SDG prevented the development of diabetes by 75%. Development of diabetes was associated with an increase in serum and pancreatic MDA, and in WBC-CL, and a decrease in pancreatic antioxidant reserve. Prevention of diabetes by SDG was associated with a decrease in serum and pancreatic MDA and WBC-CL and an increase in pancreatic antioxidant reserve. CONCLUSIONS: These results suggest that STZ-induced diabetes is mediated through oxidative stress and that SDG is effective in reducing the STZ-induced diabetes mellitus. PMID- 10839205 TI - Propionyl-L-carnitine effects on postischemic recovery of heart function and substrate oxidation in the diabetic rat. AB - Previous studies have shown that propionyl-L-carnitine (PLC) can exert cardiac antiischemic effects in models of diabetes. In the nonischemic diabetic rat heart, PLC improves ventricular function secondary to stimulation in the oxidation of glucose and palmitate. Whether this increase in the oxidation of these substrates can explain the beneficial effects of PLC in the ischemic reperfused diabetic rat heart has yet to be determined. Diabetes was induced in male Sprague-Dawley rats by an intravenous injection of streptozotocin (60 mg/kg). Treatment was initiated by supplementing the drinking water with propionyl-L-carnitine at the concentration of 1 g/L. After a 6-week treatment period, exogenous substrate oxidation and recovery of mechanical function following ischemia were determined in isolated working hearts. In aerobically perfused diabetic hearts, compared with those of controls, rates of glucose oxidation were lower, but those of palmitate oxidation were similar. Diabetes was also characterized by a pronounced decrease in heart function. Following treatment with by propionyl-L-carnitine, however, there was a marked increase in rates at which glucose and palmitate were oxidized by diabetic hearts and a significant improvement in heart performance. Postischemic recovery of function in diabetic hearts was also improved with PLC. This improvement in contractile function was accompanied by an increase in both glucose and palmitate oxidation. Our findings show that postischemic diabetic rat heart function can be improved following chronic PLC treatment. This beneficial effect of propionyl-L-carnitine can be explained, in part, by an improvement in the oxidation of glucose and palmitate. PMID- 10839206 TI - Nitric oxide effect on colonocyte metabolism: co-action of sulfides and peroxide. AB - Luminal levels of nitric oxide/nitrite are high in colitis. Whether nitric oxide is injurious or protective to human colonocytes is unknown and the role of nitric oxide in the genesis of colitis unclear. The aims were to establish whether nitric oxide was injurious to oxidation of substrates (n-butyrate and D-glucose) in isolated human and rat colonocytes both alone and in the presence of hydrogen sulfide and hydrogen peroxide, agents implicated in cell damage of colitis. Nitric oxide generation from S-nitrosoglutathione was measured by nitrite appearance. Colonocytes were isolated and incubated with [1-14C] butyrate or [6 14C] glucose and 2.6 microM nitric oxide, 1.5 mM sodium hydrogen sulfide or 2.5 mM hydrogen peroxide. Acyl-CoA esters were measured by high performance liquid chromatography, 14CO2 radiochemically and lactate/ketones by enzymic methods. Results indicate that nitric oxide very significantly (p < .001) reduced acyl-CoA formation but did not impair 14CO2 generation. Peroxide and sulfide with nitric oxide resulted in significant reduction (p < 0.01) of substrate oxidation to CO2. Sulfide significantly stimulated release of nitric oxide from S nitrosoglutathione. The principal conclusion is that nitric oxide diminishes CoA metabolism in colonocytes. CoA depletion has been observed in chronic human colitis for which a biochemical explanation has been lacking. For acute injurious action in human colonocytes nitric oxide requires co-action of peroxide and sulfide to impair oxidation of substrates in cells. From current observations treatment of colitis should aim to reduce simultaneously nitric oxide, peroxide and sulfide generation in the colon. PMID- 10839207 TI - Differential regulation of testosterone vs. 5alpha-dihydrotestosterone by selective androgen response elements. AB - There are two major physiological androgens, testosterone (T), and 5alpha dihydrotestosterone (DHT), which induce different responses in mammals. These androgens regulate the target gene transcription via binding to and activating the same androgen receptor (AR). The molecular mechanisms that differ between these two very close androgens through the same AR protein to target the distinct genomic responses remain unknown. Using yeast genetic selection, we identified two kinds of androgen response elements (ARE), which could respond differentially to T vs. DHT. These two AREs also show different T- vs. DHT-induced AR transactivation in mammalian Chinese hamster ovary (CHO) cells in terms of copy number and comparisons with the classic mouse mammary tumor virus ARE. Together, our results suggest that the selective ARE sequence may play an important role in the differential T- vs. DHT-induced AR transactivation. PMID- 10839208 TI - Anti-diabetic and toxic effects of vanadium compounds. AB - Compounds of the trace element vanadium have been shown to mimic insulin in in vitro and in vivo systems. These compounds have been found to exert anti-diabetic effects in rodent models of type 1 and type 2 diabetes mellitus as well as in a limited number of studies in human diabetic subjects. Thus, vanadium compounds have emerged as agents for potential use in diabetes therapy. However, treatment of diabetic animals with inorganic vanadium salts has also been associated with some toxic side-effects such as gastrointestinal discomfort and decreased body weight gain. In addition, vanadium salts have been reported to exert toxic effects on the liver and kidney. More recently, it was shown that organic vanadium compounds were much safer than inorganic vanadium salts and did not cause any gastrointestinal discomfort, hepatic or renal toxicity. This review briefly summarizes the anti-diabetic and toxic effects of vanadium compounds. PMID- 10839209 TI - A double-blind, placebo-controlled pilot study of the stimulating and adaptogenic effect of Rhodiola rosea SHR-5 extract on the fatigue of students caused by stress during an examination period with a repeated low-dose regimen. AB - The objective was to investigate the stimulating and normalizing effect of the adaptogen Rhodiola rosea extract SHR-5 in foreign students during a stressful examination period. The study was performed as a double-blind, randomized and placebo-controlled with low repeated dose regime. The study drug and the placebo were taken for 20 days by the students during an examination period. The physical and mental performance were assessed before and after the period, based on objective as well as on subjective evaluation. The most significant improvement in the SHR-5 group was seen in physical fitness, mental fatigue and neuro-motoric tests (p <0.01). The self-assessment of the general well-being was also significantly (p < 0.05) better in the verum group. No significance was seen in the correction of text tests or a neuro-muscular tapping test. The overall conclusion is that the study drug gave significant results compared to the placebo group but that the dose level probably was suboptimal. PMID- 10839210 TI - Endothelium-derived nitric oxide is involved in the hypotensive and vasorelaxant responses induced by the aqueous fraction of the ethanolic extract of the leaves of Albizia inopinata (Harms) G. P. Lewis in rats. AB - The acute cardiovascular effects of an aqueous fraction of the ethanolic extract of the leaves (AFL) of Albizia inopinata (Harms) G. P. Lewis (Leguminosae) were studied in rats using a combined in vivo and in vitro approach. In conscious, unrestrained rats, AFL (5, 10 and 20 mg/kg(-1) body wt. i.v., randomly) produced a significant and dose-dependent hypotension associated with increases in heart rate and cardiac output, and with a strong reduction in total peripheral resistances. The hypotensive response to AFL (20 mg/kg(-1) body wt.) was attenuated significantly after nitric oxide (NO) synthase blockade (L-NAME, 20 mg/kg(-1) body wt. i.v.). Furthermore, under these conditions, the associated tachycardia was inhibited completely. In isolated rat aortic rings, increasing concentrations of AFL (10, 20, 40 and 80 microg/ml(-1)) were able to antagonize the effects of phenylephrine- (1 microM) and KCl- (80 mM) induced contractions (IC50 value 65 +/- 4 and 54 +/- 6 microg/ml(-1), respectively). The smooth muscle relaxant activity of AFL was inhibited similarly either removal of the vascular endothelium or by L-NAME (10 and 100 microM), but was not affected significantly by atropine (1 microM) or indomethacin (10 microM). In isolated rat atrial preparations, AFL (30, 100, 300 and 500 microg/ml(-1)) produced concentration related negative inotropic and chronotropic effects (IC50 value = 274 +/- 53 and 335 +/- 23 microg/ml(-1), respectively). These results suggest that in rats, the hypotensive effect of AFL is due to a peripheral vasodilation, at least partly secondary to the release of NO by the vascular endothelium. The direct cardio depressant actions of AFL are of little importance in the systemic effects of the extract. PMID- 10839211 TI - Hypotensive action of Nangapiry, a Paraguayan natural medicine, in rodents. AB - Hypotensive action mechanism of a cation exchange resin adsorbate (IR-120A) separated from a Paraguayan Natural Medicine, Nangapiry, was investigated. Blood pressures of normal and pithed rats and contractions of isolated thoracic aorta and atria of mice were measured. The blood pressure on normal rats was reduced by an intravenous injection of IR-120A (5 mg/kg). The hypotensive effect on the pithed rat appeared more lasting than that on normal rats by IR-120A. The IR-120A (100 microg-3 mg/ml) concentration-dependently depressed prostaglandin (PG) F2alpha (10 microM)- or KCl (40 mM)-induced aortic contractions and electrically evoked contraction of left atria, and at a lesser extent spontaneous beating rate of right atria. The 50% inhibitory concentrations (IC50) for the PGF2alpha- and KCl-induced aortic contractions were 713 and 828 microg/ml, respectively, and the IC50 values for the muscle contraction and the beating rate were 1.04 and >3 mg/ml, respectively. These results suggest that the hypotensive action of IR-120A are peripherally elicited by the dilatation of artery and the depression of heart contraction, but not the reduction of heart rate. PMID- 10839212 TI - Anti-sickling, analgesic and anti-inflammatory properties of 3,5-dimethoxy-4 hydroxy benzoic acid and 2,3,4-trihydroxyacetophenone. AB - Effects of 3,5-dimethoxy-4-hydroxybenzoic acid and 2,3,4-trihydroxyacetophenone were studied on haemoglobin S (Hb S) polymerisation, analgesia and inflammation using Hb S solution, rats and mice. UV spectrophotometric procedure was used to monitor the polymerization of the Hb S. Acetic acid induced writhing in mice and egg albumin induced rat paw edema procedures were used to evaluate analgesic and anti-inflammatory activities of the compounds respectively. The results indicate that both drugs inhibit the process of polymerization significantly, possibly by direct action on the Hb S molecules. The drugs inhibited acetic acid induced pain and decreased egg albumin induced oedema. It is concluded that 3,5-dimethoxy-4 hydroxybenzoic acid and 2,3,4-trihydroxyacetophenone may have some value in the management of sickle cell disease. PMID- 10839213 TI - Analysis of the antinociceptive properties of marrubiin isolated from Marrubium vulgare. AB - We have shown previously that Marrubium vulgare, a medicinal plant employed frequently in folk medicine to treat a variety of ailments, exhibits antispasmodic and antinociceptive effects in different experimental models. This work describes the antinociceptive profile of marrubiin, the main constituent of this plant, which was analysed in some models of nociception in mice. The results showed that marrubiin exhibits potent and dose-related antinociceptive effects, whose calculated ID50 values (micromol/kg, i.p.) were the following: 2.2 in the writhing test, 6.6 (first phase) and 6.3 (second phase) in the formalin-induced pain test and 28.8 when evaluated in the capsaicin test. It was more potent than some well-known analgesic drugs. The antinociception produced by the marrubiin was not reversed by naloxone when analyzed against the writhing test. In the hot plate test, marrubiin did not increase the latency period of pain induced by the thermal stimuli. Its exact mechanism of action remains to be determined, but the results suggest that marrubiin, like hydroalcoholic extract of M. vulgare, does not interact with opioid systems. PMID- 10839214 TI - Effect of Maytenus aquifolium extract on the pharmacokinetic and antiinflammatory effectiveness of piroxicam in rats. AB - This study explored the interference by Maytenus aquifolium leaves hydroalcoholic (MALHE) extract, administered orally, on the pharmacokinetic and antiinflammatory activity of piroxicam in rats. The results showed no significant difference in piroxicam bioavailability with simultaneous application of MALHE. MALHE also had no effect on the inhibitory effect of piroxicam on inflammatory processes induced by carrageenan and complete Freund adjuvant. PMID- 10839215 TI - Anti-inflammatory, antioxidant and antimicrobial activity of Ophthacare brand, an herbal eye drops. AB - In the present study, the herbal preparation of Ophthacare brand eye drops was investigated for its anti-inflammatory, antioxidant and antimicrobial activity, using in vivo and in vitro experimental models. Ophthacare brand eye drops exhibited significant anti-inflammatory activity in turpentine liniment-induced ocular inflammation in rabbits. The preparation dose-dependently inhibited ferric chloride-induced lipid peroxidation in vitro and also showed significant antibacterial activity against Escherichia coli and Staphylococcus aureus and antifungal activity against Candida albicans. All these findings suggest that Ophthacare brand eye drops can be used in the treatment of various ophthalmic disorders. PMID- 10839216 TI - Chemopreventive effects of sage oil on skin papillomas in mice. AB - Salvia libanotica (sage) extract is a popular plant remedy used by Middle Eastern people to treat common complaints such as colds and abdominal pain. In this study, the chemopreventive effects of sage oil on 7,12 dimethylbenz[a]anthracene (DMBA)-initiated and 12-O-tetradecanoylphorbol-13-acetate (TPA)-promoted skin papillomas was investigated. Furthermore, its growth inhibitory and cytotoxic effects on a mouse papilloma-derived cell line (SP-1) were studied using 3H thymidine incorporation, cell count and trypan blue dye exclusion assays. Sage oil was either applied topically to mouse skin at concentrations of 5, 50 and 100% in acetone, injected intraperitoneally at concentrations of 4 (37 mg/ml) and 8% (75 mg/ml) in saline or given by gavage at 100% twice per week for 20 weeks, 20 minutes prior to each promotion treatment with TPA. The topically applied 100% oil extract delayed tumor appearance by 4 weeks and inhibited tumor incidence and yield by 19 and 61%, respectively, at week 20. Topical application of 50% and 5% sage oil inhibited tumor yield by 41% at week 20. Tumor weight was decreased by 75% and 80% following treatment of mouse skin with 50% and 100% oil, respectively. Intraperitoneal injections and gavage treatments failed to inhibit the promotion of tumors in mouse skin, but significantly decreased tumor weight and volume. Sage oil displayed strong growth inhibitory effects on the SP-1 papilloma derived cell line following 24 hrs of treatment with estimated IC50 of 50 microg/ml. This observed growth inhibition was due to cytostatic and not cytotoxic effects. Our results suggest that the oil extract of the sage plant has potent suppressive activities against tumor promotion in mouse skin and thus could be an effective chemopreventive agent against skin cancer. PMID- 10839217 TI - Treatment of chemotherapy-induced leukopenia in a rat model with aqueous extract from Uncaria tomentosa. AB - The Uncaria tomentosa water extracts (C-Med-100) depleted of indole alkaloids (< 0.05%, w/w) have been shown to induce apoptosis and inhibit proliferation in tumor cells in vitro and to enhance DNA repair, mitogenic response and white blood cells in vivo. In this study, the effect of C-Med-100 in the treatment of chemically induced leukopenia was evaluated in a rat model. W/Fu rats were treated first with doxorubicin (DXR) 2 mg/kg x 3 (i.p. injection at 24 hour intervals) to induce leukopenia. Twenty-four hours after the last DXR treatment, the rats were daily gavaged with C-Med-100 for 16 consecutive days. As a positive control, Neupogen, a granulocyte colony stimulator was also administered by subcutaneous injection at a dose of 5 and 10 microg/ml for 10 consecutive days. The results showed that both C-Med-100 and Neupogen treatment groups recovered significantly sooner (p < 0.05 by Duncan test) than DXR group. However, the recovery by C-Med-100 treatment was a more natural process than Neupogen because all fractions of white blood cells were proportionally increased while Neupogen mainly elevated the neutrophil cells. These results were also confirmed by microscopic examination of the blood smears. The mechanism of the C-Med-100 effect on WBC is not known but other data showing enhanced effects on DNA repair and immune cell proliferative response support a general immune enhancement. PMID- 10839218 TI - Cytotoxic effect and electrophysiological activity of (S)-bgugaine, an alkylpyrrolidine alkaloid against MRC-5 fibroblasts. AB - (+)-S-bgugaine [1], is an alkaloid prepared by enantioselective synthesis. This alkaloid is an isomer of R-bgugaine [2], an alkaloid isolated from Arisarum vulgare, an Araceae toxic plant of Morocco. The cytotoxic effect and the electrophysiological activity of (+)-S-bgugaine [1] against MRC-5 fibroblasts of (+)-S-bgugaine 1, were studied. (+)-S-bgugaine [1] showed a cytotoxic potential at 40 microg/ml against these MRC-5 cells. The electrophysiological study on MRC 5 cells was carried out using the technique of patch-clamp and showed that the activity of compound 1 involved a reduction of outward potassic current at the concentration of 100 microM (28.1 microg/ml) and was accentuated by 200 microM (56.2 microg/ml). In this study we show that S-bgugaine [1], decreases the outward potassic current. PMID- 10839219 TI - Beta-carotene prolongs survival, decreases lipid peroxidation and enhances glutathione status in transplantable murine lymphoma. AB - Carotenoids of dietary origin have recently been the subject of renewed research interest because of epidemiological evidence indicating an inverse relationship between intake of carotenoids-rich plant substances and risk of certain cancers. This study was attempted to understand the biological actions of dietary beta carotene (BC) on Dalton's lymphoma (DL), a rapidly proliferating transplantable tumor, in effecting the survival of the lymphoma-bearing mice. The glutathione (GSH) level and the extent of lipid peroxidation in the liver, kidney and brain were monitored in BC-treated (100 mg/kg food) mice transplanted with DL. These markers showed substantial alterations during the whole length of tumor progression in lymphoma-bearing mice without BC supplementation. When treated with BC, both malondialdehyde contents (evidence of lipid peroxidation) and the GSH levels in different organs were found to be closer to normal values in the initial period of tumor progression. BC-mediated protection against lipid peroxidation was maximally found to be in hepatic tissue throughout the study following DL transplantation. This was fairly reflected in the higher BC concentration in hepatic tissue of BC-treated lymphoma group compared to untreated lymphoma control. Significantly higher survival time (51-55 days) was observed in BC-treated animals in comparison to their untreated DL counterparts (35-38 days). The prolonged survival observed in the BC-supplemented animals may be attributed to the higher resistance offered by animals receiving BC towards lipid peroxidation-related tissue injury. PMID- 10839220 TI - Reduction of membrane fluidity by antibacterial sophoraflavanone G isolated from Sophora exigua. AB - Sophoraflavanone G (5,7,2',4'-tetrahydroxy-8-lavandulylflavanone) has been referred as a phytochemical with the intensive antibacterial activity. To elucidate the pharmacological mechanism underlying an antibacterial action, sophoraflavanone G was isolated from Sophora exigua, thereafter its effect on membrane fluidity was studied using model membranes and compared with less active naringenin lacking 8-lavandulyl and 2'-hydroxyl groups. Highly purified sophoraflavanone G of 0.05-5 microg/ml, corresponding to the minimum growth inhibitory concentrations against various bacteria, significantly increased fluorescence polarization of the liposomes prepared from 1,2-dipalmitoyl-L-alpha phosphatidylcholine and 1-palmitoyl-2-oleoyl-L-alpha-phosphatidylcholine. Such increases were found in both liposomes measured with two fluorescent probes to indicate an alteration of membrane fluidity in hydrophilic and hydrophobic regions, suggesting that sophoraflavanone G reduces the fluidity of outer and inner layers of membranes. Although naringenin also showed the membrane effect, it needed concentrations over 2.5 microg/ml to induce a significant reduction of membrane fluidity. Sophoraflavanone G is considered to exert an antibacterial effect by reducing the fluidity of cellular membranes. PMID- 10839221 TI - In vitro antimicrobial activity of six medicinal plants traditionally used for the treatment of dysentery and diarrhoea in Democratic Republic of Congo (DRC). AB - Twenty-four crude extracts derived from six medicinal plants highly valued as antidiarrhoeal agents in Congolese folk medicine were screened for antimicrobial activity against several enteric pathogens. The results of this study indicated that the methanolic and aqueous extracts derived from three of them (Roureopsis obliquifoliolata, Epinetrum villosum and Cissus rubiginosa) possessed prominent antibacterial activity, therefore supporting the ethnomedical uses of these species. In addition, phytochemical analysis of these medicinal plants showed that 1/6 plant sample contained alkaloids, 6/6 triterpenes and/or sterols, 4/6 flavonoids, 3/6 tannins and 5/6 saponins. Anthraquinones were not detected in any of these plants. PMID- 10839222 TI - Effect of bioactive tannoid principles of Emblica officinalis on iron-induced hepatic toxicity in rats. AB - The tannoid principles of the fruits of the plant Emblica officinalis Gaertn comprising of emblicanin A. emblicanin B, punigluconin and pedunculagin, have been reported to exhibit antioxidant activity in vitro and in vivo. In the present study, an emblicanin A (37%) and B (33%) enriched fraction of fresh juice of Emblica fruits (EOT), administered prophylactically (10, 20 and 50 mg/kg, p.o.) for 10 consecutive day, was found to inhibit acute iron overload (30 mg/kg, i.p.) hepatic lipid peroxidation and the increase of serum levels of alanine aminotransferase, aspartate aminotransferase and lactate dehydrogenase, used as markers of the induced hepatic dysfunction. A similar effect was produced by silymarin (20 mg/kg, p.o.), an antioxidant hepatoprotective agent. The results support the use of Emblica fruits for hepatoprotection in Ayurveda. PMID- 10839223 TI - The effects of wet suits on physiological and biomechanical indices during swimming. AB - The objectives of this study were to verify the effects of wet suits (WS) on the performance during 1500m swimming (V1500), on the velocity corresponding to the anaerobic threshold (VAT) and on the drag force (AD) as well as its coefficient (Cx). 19 swimmers randomly completed the following protocols on different days (with and without WS): 1) maximal performance of 1500m swimming; 2) VAT in field test, with fixed concentration of blood lactate (4 mM) and 3) determination of hydrodynamic indices (AD and Cx). The results demonstrated significant differences (p < 0.05) in the VAT (1.27 +/- 0.09; 1.21 +/- 0.06 m.s-1), and in the V1500 (1.21 +/- 0.08; 1.17 +/- 0.08 m.s-1), with and without WS, respectively. However the AD, and its Cx did not present significant differences (p>0.05) for the respective maximal speeds of swimming. In summary, we can conclude that WS allows swimmers to reach greater speeds in both, long- and short course swims. This improvement can be related to the decrease of the AD, since with higher speeds (with WS) the subjects presented the same resistance, as they did when compared to speeds without a WS. Moreover, these data suggest that the methodology used in this study to determine the Cx is unable to detect the improvement caused by WS. PMID- 10839224 TI - The safety practices of sporting clubs/centres in the city of Hume. AB - Sports injuries are a significant public health problem in Australia. However, little information is available about community level sports injuries, or about the sports safety policies and practices of community level sports organisations in Australia. The aim of this paper is to present the results of a survey of local clubs and sporting centres in the City of Hume, a local council in Victoria. This is the first reported survey of safety practices of sporting clubs/centres at the community level in Australia. Sixty-four clubs/centres participated in the survey, which involved face-to-face interviews with representatives from the participating clubs/centres. A major finding was that whilst sports bodies perform certain activities typically associated with preventing sports injuries, they often do not have formal policies or written objectives which recognise the safety of their participants as an important goal. The sports safety measures reported to be adopted by the surveyed clubs/centres included use of protective equipment, accredited coaches, sports trainers, encouraging warm-ups, modified rules for juniors and checking of playing areas and facilities for environmental hazards. The provision of first aid services (including personnel and equipment) varied across the sporting clubs/centres. The major barriers towards improving sports safety were reported to be a lack of funds, the media's attitude towards sports injuries and the role of the local council as the owner of sporting facilities. There is also a clear role for researchers to improve the dissemination of key findings from their injury prevention research in a form that can be readily used at the grass roots of sports participation. PMID- 10839225 TI - Physiotherapists' and male professional athletes' views on psychological skills for rehabilitation. AB - In the literature on the psychological aspects of rehabilitation from athletic injury, several studies in North America have focused on athletic trainers' roles as physical and psychological caregivers for injured athletes. Wiese, Weiss, and Yukelson (1991) examined the views of athletic trainers on the psychological characteristics that distinguished athletes who coped well, versus those who coped poorly with rehabilitation, and also sought athletic trainers opinions on the role of psychological skills in injury rehabilitation. This study replicated the Wiese et al. (1991) study with Australian physiotherapists and extended it to include the viewpoints of professional basketball players. The results indicated the importance attached to communication and motivation by both physiotherapists and athletes in the rehabilitation process. Both athletes and physiotherapists did not, however, think psychological skills such as relaxation or imagery techniques to be particularly useful tools in the recovery process. PMID- 10839226 TI - Efficacy of thigh protectors in preventing thigh haematomas. AB - Thigh haematomas are extremely common in Australian Rules Football (ARF). This is in contrast to contact sports overseas, the likely reason is the increased use of thigh protectors as part of team uniforms in contact sports such as American football. Thigh haematomas can have a significant impact on an athlete's performance, ranging from short term performance impairment, muscle deconditioning and compartment syndromes, to long term problems, such as career threatening myositis ossificans and possibly muscle tears. To assess the efficacy of thigh protectors made for Australian Football, a prospective study was undertaken involving two teams in the elite junior (U 18) VSFL competition in SE Australia. One team wore thigh protectors over the course of the season while the other team acted as controls and did not wear thigh protectors. The control group suffered nine thigh haematomas, while the protected group had none (p<0.01). The possible de-conditioning effect of the haematomas was evidenced by two of the control group suffering torn quadriceps within four weeks of the haematoma. The protectors were generally well tolerated by all but one player, except in hot conditions, when they were uncomfortable. PMID- 10839227 TI - The effects of a repeated bout of eccentric exercise on indices of muscle damage and delayed onset muscle soreness. AB - This study examined markers of muscle damage following a repeated bout of maximal isokinetic eccentric exercise performed prior to full recovery from a previous bout. Twenty non-resistance trained volunteers were randomly assigned to a control (CON, n=10) or experimental (EXP, n=10) group. Both groups performed 36 maximal isokinetic eccentric contractions of the elbow flexors of the non dominant arm (ECC1). The EXP group repeated the same eccentric exercise bout two days later (ECC2). Total work and peak eccentric torque were recorded during each set of ECC1 and ECC2. Isometric torque, delayed onset muscle soreness (DOMS), flexed elbow angle and plasma creatine kinase (CK) activity were measured prior to and immediately following ECC1 and ECC2. at 24h intervals for 7 days following ECC1 and finally on day 11. In both groups, all dependent variables changed significantly during the 2 days following ECC1. A further acute post-exercise impairment in isometric torque (30 +/- 5%) and flexed elbow angle (20 +/- 4%) was observed following ECC2 (p<0.05), despite EXP subjects producing uniformly lower work and peak eccentric torque values during ECC2 (p<0.05). No other significant differences between the CON and EXP groups were observed throughout the study (p>0.05). These findings suggest that when maximal isokinetic eccentric exercise is repeated two days after experiencing of contraction-induced muscle damage, the recovery time course is not significantly altered. PMID- 10839228 TI - Neutrophil oxidative activity is differentially affected by exercise intensity and type. AB - The differential effects of exercise intensity and type on neutrophil activation were assessed in eight well-trained male runners. Each subject undertook, on different days, three separate 40 min interval (8 x 5 min) treadmill bouts: an intense uphill run (90% VO2 max), a moderate-intensity near-level run and an eccentrically-biased downhill run (both at 52% VO2 max). Blood granulocyte count increased (p< 0.05) after all three treadmill bouts (range 25-108%). Chemiluminescence activity of isolated neutrophils decreased (p< 0.05) immediately after (-58%) and 1-h after (-72%) uphill running, but became significantly elevated (p< 0.05) at 6-h after the near-level (+71%) and downhill (+84%) runs. The ability of neutrophils to release the superoxide anion radical was reduced (p< 0.05) immediately after near-level (-29%) and uphill (-21%) running in cells stimulated with opsonized zymosan. Epinephrine concentration increased by 430% (p=0.01) after uphill but not with near-level or downhill running. The plasma concentration of elastase increased (p< 0.05) immediately after uphill and near-level running, and one hour after uphill running. These results suggest that a population of neutrophils mobilised into the circulation became directly activated in response to exercise, and that neutrophil oxidative activity is affected differentially by both the intensity and type of exercise undertaken. PMID- 10839229 TI - A multistage shuttle swim test to assess aerobic fitness in competitive water polo players. AB - A 10m multistage shuttle swim test (MSST) was designed for the assessment of aerobic fitness of competitive water polo players. Test-retest reliability was determined using a sample of 22 female and 22 male trained water polo players. An intraclass correlation coefficient of 0.99 (p>0.05) was calculated between the two test scores. The technical error of measurement for the test was 2.3 shuttles or 5.0%. The validity of the test was determined using a sample of 13 female and 12 male water polo players. A validation correlation coefficient of 0.88 resulted between the number of shuttles completed during the MSST and VO2max [litres/body surface area/ minute (l x BSA-1 x min-1)] measured during an incremental tethered swim test to exhaustion. A stepwise multiple regression revealed that VO2max (l x BSA-1 x min-1) accounted for approximately 78% of the MSST variance. It was concluded that the 10m multistage shuttle swim test is a reliable and valid field test of aerobic fitness for use with trained water polo players. PMID- 10839230 TI - Preventing injuries to competitive and recreational adult golfers: what is the evidence? AB - Although injuries to golfers are not common, both survey studies and hospital emergency department records have provided objective evidence that golfers are sometimes at risk of injury. While many golf injury studies describe the associated mechanisms and types of injuries, less attention has been given to research relating to the various injury prevention measures for this sport. This paper provides a critical review of the range of countermeasures to prevent golf injuries and highlights areas to be considered for future research, development, and implementation. In particular, it focuses on the strength of the evidence for the effectiveness of these measures. This review concludes that there needs to be more formal evaluation of the suggested countermeasures to prevent golf injuries. Particular attention should be given to evaluations of the golf swing and its relationship to particular injuries such as low-back, wrist, and shoulder problems. These countermeasures and associated strategies are recommended to reduce the incidence of injury in this popular and accessible sport. PMID- 10839231 TI - Beta-hydroxy beta-methylbutyrate (HMB) supplementation does not influence the urinary testosterone: epitestosterone ratio in healthy males. AB - Six healthy, recreationally active, males undertook two weeks supplementation with beta-Hydroxy beta-Methylbutyrate (HMB). Supplementation was in capsule form with 3 g consumed each day in three even doses of 1 g at main meals. Mid stream urine samples were collected prior to, as well as, after one and two weeks of supplementation and subsequently analysed for testosterone and epitestosterone. The testosterone: epitestosterone ratio was not affected by 2 weeks of HMB supplementation (mean +/- SD baseline 1.02 +/- 0.68; week one 0.98 +/- 0.61; week two 0.92 +/- 0.62). Our results support the claim that supplementation with HMB at the doses recommended will not influence the urinary testosterone: epitestosterone ratio and thus not breach doping policies of the International Olympic Committee for exogenous testosterone or precursor administration. PMID- 10839232 TI - What exercise and sport scientists don't understand. AB - The power of research design in studies published in the Australian Journal of Science and Medicine in Sport (AJSMS: now the Journal of Science and Medicine in Sport) for the years 1996 and 1997 were analysed for their ability to detect small, medium, and large effects according to Cohen's (1988) conventions. Also examined were the reporting and interpreting of effect sizes and the control for experiment-wise (EW) Type I error rates. From the two years of articles, 29 studies were analysed, and power was computed on 108 different tests of significance. The median power of the studies to detect small, medium, and large effects were .14, .65 and .97, respectively. These results suggest that exercise and sport science research, at least as represented in AJSMS, is probably underpowered and may be limited in detecting small effects, has a better, but still underpowered, chance of detecting medium effects, and has adequate power principally for detecting large effects. The reporting of effect sizes was rare, and adequate interpretation of them was even rarer. The mean EW Type I error rate for all studies was .49. The analyses conducted suggest that much research in exercise science may have substantial Type I and Type II errors. An appeal is made for exercise scientists to conduct power analyses, control for EW error, exercise caution in the interpretation of nonsignificant results, and examine, report, and interpret effect sizes rather than solely rely on p values to determine whether significant changes occurred or significant relationships exist. PMID- 10839233 TI - Comparative study of propofol or propofol and ketamine for the induction of anaesthesia in dogs. AB - The effects of propofol alone or propofol and ketamine for the induction of anaesthesia in dogs were compared. Thirty healthy dogs were premedicated with acepromazine and pethidine, then randomly allocated to either treatment. Anaesthesia was induced with propofol (4 mg/kg bodyweight intravenously) (group 1), or propofol and ketamine (2 mg/kg bodyweight of each intravenously) (group 2). Anaesthesia was maintained with halothane, delivered in a mixture of oxygen and nitrous oxide (1:2) via a non-rebreathing Bain circuit. Various cardiorespiratory parameters were monitored at two, five, 10, 15, 20, 25 and 30 minutes after induction, and the animals were observed during anaesthesia and recovery, and any adverse effects were recorded. During anaesthesia, the heart rate, but not the systolic arterial pressure, was consistently higher in group 2 (range 95 to 102 beats per minute) than in group 1 (range 73 to 90 beats per minute). Post-induction apnoea was more common in group 2 (11 of 15) than in group 1 (six of 15). Muscle twitching was observed in three dogs in each group. Recovery times were similar in both groups. Propofol followed by ketamine was comparable with propofol alone for the induction of anaesthesia in healthy dogs. PMID- 10839234 TI - Anatomical, cytogenetic and behavioural studies of freemartin ewes. AB - The external phenotype and reproductive behaviour of 21 freemartin sheep and two male pseudohermaphrodite sheep were recorded with the aim of identifying any characteristics which might be used to predict a failure to breed. The size and shape of the vulva and clitoris, the length of the vagina, the size of the teats, the presence or absence of inguinal gonads, and the ultrasonographic characteristics of the inguinal gonads were recorded. A subjective assessment of the masculinity of each animal's body form was also made, and its behavioural responses to a virile ram and to an oestrus ewe were recorded. A number of physical and behavioural abnormalities were detected but the only consistent finding in all 23 animals was a short vagina which varied in length from 3.1 to 7.0 cm, compared with 10 to 14 cm in normal animals. PMID- 10839235 TI - Study of the hyaloid apparatus in the neonatal thoroughbred foal. AB - Over 80 per cent of 169 neonatal thoroughbred foals had some part of the hyaloid artery present bilaterally and its degree of completeness, colour and character was statistically symmetrical. The complete artery was present in over 60 per cent of the foals. The incidence of the hyaloid artery and its completeness, colour and character were related to the age of the foal. The hyaloid system atrophies in the early stages of the neonatal period and generally its presence did not cause any significant ocular problems. The presence and morphology of the posterior pupillary membrane was highly correlated with the presence, completeness, colour and character of the hyaloid artery. PMID- 10839236 TI - Bovine intestinal spirochaetosis with dysentery. PMID- 10839237 TI - Natural case of bovine herpesvirus 1 meningoencephalitis in an adult cow. PMID- 10839238 TI - Isolations of H1N2 influenza A virus from pigs in Belgium. PMID- 10839239 TI - Rabies vaccinations. PMID- 10839240 TI - Scouring in lambs following treatment with Vecoxan. PMID- 10839241 TI - Porcine circovirosis in Spain. PMID- 10839242 TI - Unseasonal snake bites in dogs. PMID- 10839243 TI - Bovine TB. PMID- 10839244 TI - The impact of hospital restructuring on social work services: a case study of a large, university-affiliated hospital in Canada. AB - The study investigates the impact of organizational restructuring on social work practice at a large, university-affiliated hospital. The article first reviews relevant literature on the patient-focused care model and presents an overview of the study setting. Several hypotheses are then presented that informed the research process. Workload measurement, interview, and questionnaire data are analyzed to test these hypotheses. The results indicate that the shift to program management has had some intended effects, particularly regarding increases in direct services supporting patients. Social workers reported somewhat lower levels of job satisfaction than expected, which partly reflects the dismantling of the professional infrastructure. Finally, the authors discuss implications for the future of the social work profession under the program management model. PMID- 10839245 TI - Community-dwelling cognitively impaired frail elders: an analysis of social workers' decisions concerning support for autonomy. AB - Home health care practice with cognitively impaired elders and their families often raises difficult clinical and ethical issues for social workers. This descriptive study examined the influence of safety, caregiver burden, and diagnostic labels on social workers' support for autonomy. All three factors were found to be significantly related to support for autonomy. There were also interactive effects between safety, caregiver burden and diagnosis. Especially important was the identification of the influence of diagnostic labels on social workers' support for autonomy. Respondents supported less autonomy for elders depicted as diagnosed with Alzheimer's disease than those diagnosed with stroke. PMID- 10839246 TI - Understanding variation in resident needs and services in homes for adults. AB - Long term care in the U.S. has been influenced greatly by the emergence and growth of alternatives to traditional nursing homes. Adult homes provide personal care and assistance to an increasing number of elderly and represent a growth sector of long term care services. However, adult homes have not been discussed widely in the healthcare literature. This paper summarizes findings from a study of adult homes and profiles their operating characteristics, services, and resident needs. Findings indicate differences between licensed and unlicensed homes for adults in terms of the needs of their residents, use of external resources and referral to other sources of care. Implications for coordination of care for the elderly are addressed. PMID- 10839247 TI - When the care giver needs care. AB - Case studies are used to illustrate family dysfunction that can occur when the wife and/or mother who has assumed the role of primary care giver is diagnosed and treated for cancer. These women were treated by the liaison psychiatrist and social workers at a large cancer center. The cases are discussed from a framework of family systems theory. Treatment implications for social workers in health care are also presented. PMID- 10839248 TI - Advancing social work practice in end-of-life care. AB - Insufficient training of health professionals has often been cited as a major barrier to improving the system of care for dying patients and for the bereaved. Although specific problems have been identified for physicians and nurses, the problems of social work in this substantive area have only recently been explored. This study used a practitioner survey, focus groups, and a survey of faculty of schools of social work to broaden the information base. Results suggested that not unlike the professions of medicine and nursing, social work knowledge and skill development in the care of the dying is uneven and not integrated sufficiently with theoretical concepts and research. Social workers felt unprepared for this work by their master's level training and unsupported by continuing education programs. They recognized few social work scholars who could function as role models by providing comprehensive training, knowledge building, innovation, and advocacy. A program for leadership development was created to test new approaches to professional development in the care of the dying and the bereaved. PMID- 10839249 TI - Imaging correlates of successful endoscopic third ventriculostomy. AB - OBJECT: The goal of this study was to determine and compare imaging correlates in pediatric patients who underwent successful or failed endoscopic third ventriculostomies (ETVs). To this end, the authors measured ventricular size changes and the presence of cerebrospinal fluid (CSF) flow void in both groups of children following ETV. METHODS: Images obtained in children with hydrocephalus immediately before and at least 30 days after having undergone ETV were reviewed by four independent observers (two blinded and two nonblinded). Each observer independently measured the frontal and occipital horn ratio ([FOR], a reliable and valid measure of ventricular size) and provided a subjective assessment of the presence of a flow void at the ETV site, the degree of periventricular edema, and the amount of CSF over the cerebral hemispheres. There were 29 children whose mean age was 6.6 years at the time of ETV and who had a mean postoperative follow up period lasting 1.6 years. Postoperatively, the mean reduction in ventricular size (as measured using the FOR) was 7% (95% confidence interval [CI] 3-11%) in cases that were deemed failures (eight patients) and 16% (95% CI 12-20%) in clinically successful cases (21 patients). This reduction was significantly greater in cases of clinical success compared with those that were deemed failures (p = 0.03, t-test). There were no substantial differences between blinded and nonblinded assessments. Flow void was present in 94% of successes and absent in 75% of failures (p = 0.01, Fisher's exact test). The other subjective assessments were not significantly different between the groups of successes and failures. CONCLUSIONS: Ventricular size appears to be somewhat reduced in both groups of patients who underwent clinically successful and failed ETV; however, the reduction is significantly greater among clinically successful cases. The presence of a flow void also appears to correlate with clinical success and its absence with clinical failure. PMID- 10839250 TI - Asymptomatic Chiari Type I malformations identified on magnetic resonance imaging. AB - OBJECT: Chiari Type I malformation (CMI) is a congenital disorder recognized by caudal displacement of the cerebellar tonsils through the foramen magnum and into the cervical canal. Frequently, associated findings include abnormalities of nearby bony and neural elements as well as syringomyelia. Cerebellar tonsillar ectopia is generally considered pathological when greater than 5 mm below the foramen magnum. However, asymptomatic tonsillar ectopia is an increasingly recognized phenomenon, the significance of which is poorly understood. METHODS: The authors retrospectively reviewed the records of all brain magnetic resonance (MR) images obtained at our hospital over a 43-month period in an attempt to ascertain the relative prevalence and MR imaging characteristics of asymptomatic CMIs. Of 22,591 patients who underwent MR imaging of the head and cervical spine, 175 were found to have CMIs with tonsillar herniation extending more than 5 mm below the foramen magnum. Of these, 25 (14%) were found to be clinically asymptomatic. The average extent of ectopia in this population was 11.4 +/- 4.86 mm, and was significantly associated with a smaller cisterna magna. Syringomyelia and osseous anomalies were found in only one asymptomatic patient. CONCLUSIONS: The authors suggest that the isolated finding of tonsillar herniation is of limited prognostic utility and must be considered in the context of all available clinical and radiographic data. Strategies for treating patients with asymptomatic CMIs are discussed. PMID- 10839251 TI - In vivo intracranial pressure dynamics in patients with hydrocephalus treated by shunt placement. AB - OBJECT: With the commercial availability of a variety of shunt systems, there is considerable controversy over the choice of the most appropriate shunt valve for each individual with hydrocephalus. Although the performance characteristics of all shunt systems are well documented in the laboratory setting, there is little description of the in vivo dynamics of intracranial pressure (ICP) after implantation of commonly used shunt systems in humans. The authors coupled telemonitoring devices to several different shunt systems to measure the performance characteristics of these valve systems with respect to intraventricular pressure (IVP) at increments of head elevation. METHODS: Twenty five patients with different shunt systems and three control patients without shunts were studied for IVP at 0 degrees, 15 degrees, 30 degrees, 45 degrees, 60 degrees, 75 degrees, and 90 degrees of head elevation, and the resultant curves were analyzed for the best-fit regression coefficient. For purposes of analysis the authors grouped shunt valve systems by design characteristics into three groups: differential-pressure valves (r = -0.321 +/- 0.061; 11 patients), nonsiphoning systems (r = -0.158 +/- 0.027; 10 patients), and flow-regulated valves (r = -0.16 +/- 0.056; four patients); there were three control patients without shunts (r = -0.112 +/- 0.037). CONCLUSIONS: The authors found that differential-pressure valves always caused ICP to drop to 0 by 30 degrees of head elevation, whereas all other valve systems caused a more gradual drop in ICP, more consistent with pressures observed in the control patients without shunts. Not surprisingly, the differential-pressure valve group was found to have a significant difference in mean regression coefficient when compared with those in whom nonsiphoning shunts (p < 0.023) or no shunts were placed (p < 0.049). These data provide a basis for evaluating shunt valve performance and for predicting valve appropriateness in patients in whom characteristics such as pressure and flow dynamics are weighed in the choice of a specific valve for implantation. PMID- 10839252 TI - Pathophysiology of long-standing overt ventriculomegaly in adults. AB - OBJECT: Long-standing overt ventriculomegaly in adults (LOVA) is a unique form of hydrocephalus that develops during childhood and manifests symptoms during adulthood. The aim of the present study was to analyze the specific pathophysiological characteristics of LOVA. METHODS: The specific diagnostic criteria for LOVA include severe ventriculomegaly in adults that is associated with macrocephalus measuring more than two standard deviations in head circumference and/or neuroradiological evidence of a significantly expanded or destroyed sella turcica. Twenty patients who fulfilled these criteria, 14 males and six females, were retrospectively studied. These patients' ages at diagnosis ranged from 15 to 61 years (mean 39.4 years). All had symptoms and/or signs indicating that hydrocephalus first occurred at birth or during infancy in the absence of any known underlying disease. The authors performed a pathophysiological study that included specific variations of magnetic resonance (MR) imaging, such as fluid-attenuated inversion recovery and cardiac-gated cine mode imaging; intracranial pressure (ICP) monitoring; three-dimensional computerized tomography (CT) scanning; and other techniques. Hydrocephalus was caused by aqueductal stenosis in all patients. Severe ventriculomegaly involving the lateral and third ventricles was associated with a marked expansion or destruction of the sella turcica in 17 cases. Cardiac-gated cine-MR imaging did not reveal any significant movements of cerebrospinal fluid in the aqueduct. Three-dimensional CT ventriculography confirmed that the expanded third ventricle protruded into the sella and, sometimes, extended a diverticulum. Fourteen patients revealed symptoms and signs that indicated increased ICP with prominent pressure waves. Dementia or mental retardation was seen in 11 patients, gait disturbance in 12, and urinary incontinence in eight; all three of these symptoms were observed in seven patients. Thirteen patients experienced visual disturbance. Nine patients underwent ventriculoperitoneal shunt implantation as the initial treatment, leading to postoperative subdural hematoma in all seven cases in which a differential pressure valve was used. Nine patients, three of whom were initially treated by shunt placement, underwent a neuroendoscopic procedure, mainly for third ventriculostomy. Postoperatively, ICP returned to normal, and marked to-and-fro pulsatile movements at the site of ventriculostomy were recognized on cine-MR imaging in patients treated endoscopically. However, the ventriculomegaly was little improved. Consequently, all patients eventually demonstrated improvement in response to either a shunt equipped with a pressure programmable valve or an endoscopic procedure; however, depression appeared in six patients, who required psychiatric consultation or medication. CONCLUSIONS: Such remarkably decreased intracranial compliance but relatively high ICP dynamics are the pathophysiological characteristics of LOVA. The therapeutic regimen should be determined based on the individual's specific pathophysiological makeup. PMID- 10839253 TI - Seven years of clinical experience with the programmable Codman Hakim valve: a retrospective study of 583 patients. AB - OBJECT: The goal of this study was to assess the value of the Codman Hakim programmable valve to settings in the range of 30 to 200 mm H2O. This valve can be adjusted noninvasively for cerebrospinal fluid (CSF) drainage. METHODS: The authors conducted a single-center retrospective study of 583 patients (421 adults and 162 children) suffering from hydrocephalus of various causes (379 patients), normal-pressure hydrocephalus (174 patients), arachnoid cyst (14 patients), and pseudotumor cerebri (16 patients). In all cases a Codman Hakim programmable valve was implanted; in 82.8% of cases it was included during the patient's first shunt implantation. In 42.4% of the cases valve pressure adjustment was required at least once (mean number of adjustments 1.2, maximum 23). The patients' clinical status improved after 64.6% of pressure adjustments. Accidental resetting of opening pressure, other than that caused by magnetic resonance (MR) imaging, was uncommon. Because MR imaging caused resetting in 26.8% of cases in which it was used, it was deemed mandatory to obtain an x-ray film after MR imaging. Valve malfunction, blockage, or adjustment difficulties occurred in 2% of valves implanted, and nontraumatic subdural fluid collections were demonstrated in 5.1% of patients (13 of whom were treated by valve pressure adjustment alone). Five year shunt survival was 53.1% for first-time shunt implantations. The shunt infection rate was 8.5% of valve implantations. Catheter-related complications and shunt-related infections were the main reasons for surgical revision and the major cause of shunt failure. At follow-up review, 97% of children and 90% of adults had improved. CONCLUSIONS: Because one cannot know in advance which case will turn out to be complicated, the authors' preference is to use the Codman Hakim programmable valve for all conditions in which CSF should be drained. PMID- 10839254 TI - Intracerebral steal phenomenon associated with global hyperemia in moyamoya disease during revascularization surgery. AB - OBJECT: The collateral vessels in moyamoya disease appear to retain their ability to constrict during hypocapnia but not to dilate during hypercapnia. It has been claimed that hypercapnia, as well as hypocapnia, decreases the blood flow in regions perfused by collateral vessels, presumably because of intracerebral steal. If this holds true, the decrease in blood flow may be proportional to the global hyperemia in the brain. To establish appropriate hemodynamic control during revascularization surgery, the authors monitored the jugular bulb oxygen saturation (SjO2) intraoperatively, a method that could sensitively detect global hyperemia. METHODS: A total of 17 patients, most of whom presented with transient ischemic attacks or fluctuating neurological deficits, underwent intraoperative monitoring of their SjO2 and end-tidal carbon dioxide (ETCO2) after a state of anesthesia had been induced with isoflurane (Group 1) or propofol (Group 2). In eight of these patients, the regional cerebral blood flow (rCBF) of the collateral vessel territory was also monitored by laser Doppler flowmetry during the period of cortical exposure, and a total of 113 data sets (averaged values during 2.5-minute intervals) was collected. There was fluctuation in the ETCO2 levels ranging from 36 to 44 mm Hg. The mean SjO2 level was clearly greater (p < 0.01) in Group 1 (71.8 +/- 2.2%) than in Group 2 (63.3 +/- 2.1%). An episodic fall in rCBF was observed in association with a transient increase in SjO2. Such an event was not uncommon in Group 1 and there was a greater risk of rCBF decreasing when SjO2 exceeded a cutoff level of 76% (p < 0.01). This level could sometimes be reached at a broad range of ETCO2 readings (37-44 mm Hg). In Group 2, similar events sometimes occurred when SjO2 increased beyond 70%. However, this level could be reached only with a higher ETCO2 (42-44 mm Hg). The rCBF level was negatively correlated to SjO2 (p < 0.01), but not always to ETCO2, indicating that the episodic fall in rCBF was closely related to global hyperemia rather than the absolute level of hypercapnia. CONCLUSIONS: The observed association between a fall in rCBF and global hyperemia supports the intracerebral steal hypothesis and indicates that it is prudent to avoid excessive global hyperemia. The optimal range of CO2 for isoflurane is more restricted than that for propofol, presumably because isoflurane induces hyperemia by itself. Monitoring of SjO2 appears to represent the most practical technique for detecting global hyperemia as well as global ischemia, both of which may cause ischemic complications in moyamoya disease. PMID- 10839255 TI - Preoperative embolization of cerebral arteriovenous malformations with silk sutures: analysis and clinical correlation of complications revealed on computerized tomography scanning. AB - OBJECT: The aim of this study was to determine the incidence and clinical significance of complications related to preoperative embolization of cerebral arteriovenous malformations (AVMs) with silk sutures as documented on postprocedure computerized tomography (CT) scans. METHODS: The CT scans were obtained within 12 to 24 hours after 221 (96%) of 230 consecutive embolizations in 70 patients. These CT scans were evaluated for the presence of ischemia, infarction, hemorrhage, or contrast agent extravasation. Adverse patient outcomes were determined after each embolization and were correlated with CT findings. New abnormalities demonstrated on CT scans were also correlated with the Spetzler Martin AVM grade, degree of arteriovenous shunting, and location. New abnormalities, the majority of them infarcts, resulted from 29 (13%) of 221 embolization procedures. In 11 (38%) of 29 cases of new CT findings, patients were asymptomatic, including 10 with new infarcts on CT scans. New neurological deficits occurred in 20 (8.7%) of 230 total embolization procedures in 19 patients, including one death. Permanent deficits occurred in nine patients (3.9% per embolization procedure, 12.8% per patient). Of the patients with new neurological deficits, 18 (90%) of 20 embolization procedures resulted in new abnormalities on CT scans. Two patients with new transient neurological deficits had no new findings on CT scans. Spetzler-Martin grade, AVM location, degree of arteriovenous shunting, and higher numbers of procedures were not statistically associated with a higher incidence of abnormalities on CT scans or new permanent neurological deficits. CONCLUSIONS: Silk sutures are an effective and relatively safe embolic agent. After brain AVM embolization with silk sutures, new abnormalities were found on CT scans obtained in one of eight procedures. When a new CT finding occurred, the patient had roughly equal chances of having no new symptoms, having new transient neurological deficits, or having new permanent neurological deficits. PMID- 10839256 TI - Repeated radiosurgery for incompletely obliterated arteriovenous malformations. AB - OBJECT: The goal of this study was to define treatment results of repeated arteriovenous malformation (AVM) radiosurgery, namely AVM obliteration and complications. METHODS: The authors analyzed their experience with repeated AVM radiosurgery performed in 41 patients for whom follow-up review lasted at least 2 years. The median duration of follow up was 34 months (range 7-65 months) after repeated radiosurgery in this group. The residual nidus was located within the area of focus (in field) of the initial radiosurgery in 28 patients (68%). Initial doses to the margin varied from 12.5 to 20 Gy (median 18 Gy). During repeated treatment the dose to the margin varied from 12.5 to 20 Gy (median 17 Gy) and the retreated volumes ranged from 0.4 to 7 cm3 (median 2.1 cm3). Follow up angiography performed at least 2 years postradiosurgery revealed complete AVM obliteration in 21 (70%) of 30 patients. The estimated overall 2-year obliteration rate, based on findings on magnetic resonance imaging (eight of 11 obliterated) and angiography (29 of 41 obliterated) was 71%. Obliteration rates correlated with margin doses (p = 0.0045) with a trend toward higher rates in cases with in-field nidus persistence (p = 0.0637). The dose-response curve for AVM nidus obliteration was not significantly different from that of the initial radiosurgery. In two patients (5%) intracranial AVM hemorrhage developed within 125.9 risk years after repeated radiosurgery (1.6% per patient year). Persistent symptomatic adverse radiation effects developed in two (5%) of 41 patients following repeated radiosurgery. Postradiosurgical imaging changes were identified in 11 (27%) of 41 patients, which correlated with a 12-Gy volume from repeated surgery (p = 0.019). CONCLUSIONS: When necessary, repeated AVM radiosurgery achieves obliteration with an acceptable risk. Despite the effects of previous irradiation, repeated radiosurgery required similar or slightly higher radiation doses to achieve the same in-field obliteration rates as those needed to obliterate an AVM that had not been treated by radiation previously. PMID- 10839257 TI - Adverse effects of limited hypotensive anesthesia on the outcome of patients with subarachnoid hemorrhage. AB - OBJECT: This study was aimed at clarifying the effect of intraoperative hypotensive anesthesia on the outcome of early surgery in patients with subarachnoid hemorrhage (SAH) caused by saccular cerebral aneurysms. Other factors were also screened for possible effects on the outcome. METHODS: Hospital charts in 84 consecutive patients with SAH who underwent aneurysm clipping by Day 4 were examined. Possible factors affecting the outcome were analyzed using multiple logistic regression with the dichotomous Glasgow Outcome Scale score as the outcome variable. The relationship between the intraoperative hypotension and the occurrence and severity of vasospasm was studied using both single- and multivariate analyses. CONCLUSIONS: Intraoperative hypotension had a significantly adverse effect on the outcome of SAH. Hypotension was also related to more frequent and severe manifestations of vasospasm. A long-lasting effect of brain retraction is possibly the cause of this phenomenon. The data contained in this study preclude the use of intraoperative hypotension even in a limited form. PMID- 10839258 TI - Intrasellar chordomas mimicking pituitary adenoma. AB - OBJECT: Whereas chordomas involving the sellar region are uncommon, largely or entirely intrasellar examples are rare. The goal in this study was to present examples of these rare tumors as a guide to their proper diagnosis and treatment. METHODS: The authors report three cases in which the chordomas filled the pituitary fossa and presented as nonfunctioning pituitary adenomas. All lesions exhibited the typical histological patterns and immunophenotype of chordoma. One tumor, studied ultrastructurally and subjected to DNA analysis, was shown to have a diploid histogram. The authors present a clinicopathological study of these three cases and review the literature on intrasellar chordomas. CONCLUSIONS: Although these tumors are easily misdiagnosed and therefore may not receive optimal treatment, aggressive surgical resection can yield a favorable prognosis in lesions with a limited extent. PMID- 10839259 TI - Long survival and therapeutic responses in patients with histologically disparate high-grade gliomas demonstrating chromosome 1p loss. AB - OBJECT: Allelic loss of chromosome 1p is a powerful predictor of tumor chemosensitivity and prolonged survival in patients with anaplastic oligodendrogliomas. Chromosome 1p loss also occurs in astrocytic and oligoastrocytic gliomas, although less commonly than in pure oligodendroglial tumors. This observation raises the possibility investigated in this study that chromosome 1p loss might also provide prognostic information for patients with high-grade gliomas with astrocytic components. METHODS: The authors report on seven patients with high-grade gliomas composed of either pure astrocytic or mixed astrocytic-oligodendroglial phenotypes, who had remarkable neuroradiological responses to therapy or unexpectedly long survivals. All of the tumors from these seven patients demonstrated chromosome 1p loss, whereas other genetic alterations characteristic of high-grade gliomas (p53 gene mutations, EGFR gene amplification, chromosome 10 loss, chromosome 19q loss, or CDKN2A/p16 deletions) were only found in occasional cases. The authors also assessed the frequency of chromosome 1p loss in a series of anonymous high-grade astrocytoma samples obtained from a tumor bank and demonstrate that this genetic change is uncommon, occurring in only 10% of cases. CONCLUSIONS: Although any prognostic importance of chromosome 1p loss in astrocytic or mixed astrocytic oligodendroglial gliomas can only be determined in larger and prospective series, these findings raise the possibility that some high-grade gliomas with chromosome 1p loss, in addition to pure anaplastic oligodendrogliomas, may follow a more favorable clinical course. PMID- 10839260 TI - Hydrocephalus: comparison of clipping and embolization in aneurysm treatment. AB - OBJECT: In this retrospective study conducted at Atkinson Morley's Hospital and Middlesbrough General Hospital, the authors analyzed 100 matched patients who had suffered subarachnoid hemorrhage (SAH) to determine whether the technical procedure by which aneurysms are treated affects the development of chronic hydrocephalus. METHODS: Four hundred seventy-five patients presented with SAH between 1995 and 1998. Exclusion criteria included posterior circulation aneurysms, multiple aneurysms, electively clipped or embolized aneurysms, angiographically undetected SAH, patients who died within 1 month of neurosurgical intervention, and patients with the same aneurysm location but a different Fisher grade. The authors matched 50 patients who underwent embolization of their aneurysms with another 50 who had similar Fisher grades and aneurysm types and underwent clipping of their aneurysms. The maximum incidence of ruptured aneurysms occurred in patients who were between 41 and 60 years of age, with women preponderant in both study groups. In each group, 27 patients had anterior communicating artery aneurysm, 13 had posterior communicating artery aneurysm, seven had middle cerebral artery aneurysm, and three had internal carotid artery aneurysm. The lesions in three patients in each group were Fisher Grade I, in 23 patients they were Fisher Grade II, in 14 they were Fisher Grade III, and 10 patients had Fisher Grade IV SAH. Nine patients among those with clipped aneurysms and eight of the patients who underwent embolization had hydrocephalus for which they needed intervention. These interventions included lumbar puncture, ventricular drainage, and ventriculoperitoneal (VP) shunt placement; three patients in each group needed VP shunt placement. CONCLUSIONS: The technical procedure used to treat aneurysms, whether clipping or embolization, does not significantly affect the development of chronic hydrocephalus. However, a larger sample of patients is needed for accurate comparisons and stronger conclusions. PMID- 10839261 TI - Clinical significance of pedicle aneurysms on feeding vessels, especially those located in infratentorial arteriovenous malformations. AB - OBJECT: The diminishing threshold for the application of neuroimaging leads to an increasingly frequent diagnosis of previously asymptomatic arteriovenous malformations (AVMs). In such a context, it is warranted to define the criteria that make a lesion potentially hazardous so that neurosurgeons and patients reach a decision concerning how to manage the AVM. In addition to the proposed grading system for AVMs, which assesses the risk of an actual treatment procedure, several studies have been concerned with the evaluation of angioarchitectural features. The goal of the present study is to demonstrate the significance of feeding vessel pedicle aneurysms, especially those found in infratentorial AVMs. METHODS: To determine the incidence of associated aneurysms, the authors reviewed an unselected cohort of 242 consecutive patients with AVMs managed between 1989 and 1999. Within this group were 240 patients who were treated by surgery, endovascular techniques, or a combination of both. Of these patients, 216 harbored a supratentorial and 24 an infratentorial AVM. Two additional patients with supratentorial AVMs underwent treatment of ruptured aneurysms without treatment of the AVMs. In six of the patients with supratentorial AVMs, proximal flow-related aneurysms were found on major feeding arteries, only one of which had caused hemorrhage. In only one patient were there additional distal feeding vessel pedicle aneurysms near the AVM, one of which had caused a major hemorrhage. In contrast, four of 24 patients with infratentorial AVMs had distal feeding artery pedicle aneurysms. Three of these aneurysms had caused hemorrhage. CONCLUSIONS: Pedicle aneurysms on feeding vessels are frequently associated with hemorrhage (four of five cases in this series). In our cohort of 242 treated patients (240 treated for AVM and two for an aneurysm), feeding vessel pedicle aneurysms appear to occur more frequently in conjunction with infratentorial AVMs, which justifies aggressive management to prevent incidences of morbidity associated with rupture of the aneurysm. PMID- 10839262 TI - Dependence of efficient adenoviral gene delivery in malignant glioma cells on the expression levels of the Coxsackievirus and adenovirus receptor. AB - OBJECT: Recombinant adenovirus is used as a competent vector in a wide spectrum of cancer gene therapies because of its high efficiency in gene delivery. To study the feasibility of gene therapy in malignant gliomas, the authors examined the antiproliferative effect of the adenovirally transduced wild-type p53 tumor suppressor gene by using 15 different high-grade glioma cell lines. METHODS: Although growth suppression in association with a high adenoviral p53 transduction efficiency was seen in five of 15 cell lines, it was not observed in the remaining 10 cell lines. To clarify the underlying mechanism, we examined the expression levels of the Coxsackievirus and adenovirus receptor (CAR), which is the primary receptor for adenovirus, and of the integrins alpha vbeta3 and alpha vbeta5, which promote adenoviral internalization. The expression level of the CAR gene showed a close correlation to adenoviral gene transduction efficiency in the tested cell lines, whereas the expression levels of the integrins did not. The CAR expression was decreased by wild-type p53 transduction in U251MG cells harboring mutant p53 and increased by antisense inhibition of p53 in LN443 cells with endogenous wild-type p53. CONCLUSIONS: The results of this study indicate that CAR expression is a critical determinant of transduction efficiencies in adenovirus-based gene therapy for human malignant gliomas. PMID- 10839263 TI - Improvement in cerebral blood flow and metabolism following subarachnoid hemorrhage in response to prophylactic administration of the hydroxyl radical scavenger, AVS, (+/-)-N,N'-propylenedinicotinamide: a positron emission tomography study in rats. AB - OBJECT: The hydroxyl radical scavenger (+/-)-N,N'-propylenedinicotinamide (AVS) has been shown to ameliorate the occurrence of vasospasm following experimental subarachnoid hemorrhage (SAH) and to reduce the incidence of delayed ischemic neurological deficits (DINDs) in patients with SAH. The authors investigated whether prophylactic administration of AVS could improve cerebral blood flow (CBF) and cerebral glucose utilization (CGU) following SAH in rats. METHODS: Anesthetized rats were subjected to intracisternal injection of blood (SAH group) or saline (control group). Either AVS (1 mg/kg/min) or saline (vehicle group) was continuously injected into the rat femoral vein. Forty-eight hours later, positron emission tomography scanning was used with the tracers 15O-H2O and 18F-2 fluoro-D-glucose to analyze quantitatively CBF and CGU, respectively, in the frontoparietal and occipital regions (12 regions of interest/group). In SAH rats receiving only vehicle, CBF decreased significantly (p < 0.05, Tukey's test) and CGU tended to decrease, compared with values obtained in control (non-SAH) rats receiving vehicle. In rats that were subjected to SAH, administration of AVS significantly (p < 0.05, Tukey's test) improved CBF and CGU in both the frontoparietal and occipital regions compared with administration of vehicle alone. CONCLUSIONS: Prophylactic administration of AVS improves CBF and CGU in the rat brain subjected to SAH, and can be a good pharmacological treatment for the prevention of DINDs following SAH. PMID- 10839264 TI - Complement activation in the brain after experimental intracerebral hemorrhage. AB - OBJECT: Brain edema formation following intracerebral hemorrhage (ICH) appears to be partly related to erythrocyte lysis and hemoglobin release. Erythrocyte lysis may be mediated by the complement cascade, which then triggers parenchymal injury. In this study the authors examine whether the complement cascade is activated after ICH and whether inhibition of complement attenuates brain edema around the hematoma. METHODS: This study was divided into three parts. In the first part, 100 microl of autologous blood was infused into the rats' right basal ganglia, and the animals were killed at 24 and 72 hours after intracerebral infusion. Their brains were tested for complement factors C9, C3d, and clusterin (a naturally occurring complement inhibitor) by using immunohistochemical analysis. In the second part of the study, the rats were killed at 24 or 72 hours after injection of 100 microl of blood. The C9 and clusterin proteins were quantitated using Western blot analysis. In the third part, the rats received either 100 microl of blood or 100 microl of blood plus 10 microg of N acetylheparin (a complement activation inhibitor). Then they were killed 24 or 72 hours later for measurement of brain water and ion contents. It was demonstrated on Western blot analysis that there had been a sixfold increase in C9 around the hematoma 24 hours after the infusion of 100 microl of autologous blood. Marked perihematomal C9 immunoreactivity was detected at 72 hours. Clusterin also increased after ICH and was expressed in neurons 72 hours later. The addition of N-acetylheparin significantly reduced brain edema formation in the ipsilateral basal ganglia at 24 hours (78.5 +/- 0.5% compared with 81.6 +/- 0.8% in control animals, p < 0.001) and at 72 hours (80.9 +/- 2.2% compared with 83.6 +/- 0.9% in control animals, p < 0.05) after ICH. CONCLUSIONS: It was found that ICH causes complement activation in the brain. Activation of complement and the formation of membrane attack complex contributes to brain edema formation after ICH. Blocking the complement cascade could be an important step in the therapy for ICH. PMID- 10839265 TI - Histopathological study of venous aneurysms in patients with dural arteriovenous fistulas. AB - OBJECT: Of all intracranial dural arteriovenous fistulas (DAVFs), those with cortical venous drainage associated with cortical venous ectasia or varices are predisposed to an aggressive course and produce progressive neurological symptoms or hemorrhages. The authors undertook a histological examination of venous aneurysms and arterialized veins in the proximity of these aneurysms that had been surgically removed in patients with DAVFs. METHODS: Surgical specimens were obtained in eight patients. The excised venous aneurysms and the arterialized veins in their proximity were stained using hematoxylin and eosin, van Gieson's elastic, and Masson's trichrome stain. Immunostaining was also performed for alpha smooth-muscle actin, desmin, and factor VIII antigen. Five of the patients had presented with venous hypertension, and three had intracranial hemorrhages. The arterialized vein obtained in the proximity of the venous aneurysm exhibited local irregular intimal thickening; the internal elastic lamina (IEL) was grossly preserved. All venous aneurysms in patients with venous hypertension manifested medial thickening and local intimal thickening with loss of IEL; the thickness of the wall was relatively uniform. In contrast, the wall thickness of venous aneurysms in patients with hemorrhage was extremely irregular and there was no clear delineation between the media and the intima. In media with complete disappearance of IEL, there was scant muscle tissue. CONCLUSIONS: Degenerative changes in venous aneurysms in patients with hemorrhage were much greater than in patients with venous hypertension, possibly because hemorrhages result from a more complicated interplay of anatomical, hemodynamic, and degenerative factors. PMID- 10839266 TI - Resection of suprasellar tumors by using a modified transsphenoidal approach. Report of four cases. AB - Generally accepted contraindications to using a transsphenoidal approach for resection of tumors that arise in or extend into the suprasellar region include a normal-sized sella turcica, normal pituitary function, and adherence of tumor to vital intracranial structures. Thus, the transsphenoidal approach has traditionally been restricted to the removal of tumors involving the pituitary fossa and, occasionally, to suprasellar extensions of such tumors if the sella is enlarged. However, conventional transcranial approaches to the suprasellar region require significant brain retraction and offer limited visualization of contralateral tumor extension and the interface between the tumor and adjacent structures, such as the hypothalamus, third ventricle, optic apparatus, and major arteries. In this paper the authors describe successful removal of suprasellar tumors by using a modified transsphenoidal approach that circumvents some of the traditional contraindications to transsphenoidal surgery, while avoiding some of the disadvantages of transcranial surgery. Four patients harbored tumors (two craniopharyngiomas and two hemangioblastomas) that arose in the suprasellar region and were located either entirely (three patients) or primarily (one patient) within the suprasellar space. All patients had a normal-sized sella turcica. Preoperatively, three of the four patients had significant endocrinological deficits signifying involvement of the hypothalamus, pituitary stalk, or pituitary gland. Two patients exhibited preoperative visual field defects. For tumor excision, a recently described modification of the traditional transsphenoidal approach was used. Using this modification, one removes the posterior portion of the planum sphenoidale, allowing access to the suprasellar region. Total resection of tumor was achieved (including absence of residual tumor on follow-up imaging) in three of the four patients. In the remaining patient, total removal was not possible because of adherence of tumor to the hypothalamus and midbrain. One postoperative cerebrospinal fluid leak occurred. Postoperative endocrinological function was worse than preoperative function in one patient. No other new postoperative endocrinological or neurological deficits were encountered. This study demonstrates the feasibility of using a modified transsphenoidal approach for resection of certain suprasellar, nonpituitary tumors. PMID- 10839267 TI - Neuroendoscopic aqueductal stent placement procedure for isolated fourth ventricle after ventricular shunt placement. Case report. AB - Isolated fourth ventricle (IFV) is a rare complication in patients who undergo shunt placement, and it is not easily corrected by surgical procedures. The authors report a case of IFV that was successfully treated with an aqueductal stent placed under direct visualization by using a neuroendoscope. This 36-year old suffered meningitis after partial resection of a brainstem pilocytic astrocytoma, and subsequently developed hydrocephalus for which a ventriculoperitoneal shunt was placed. Nine months later, the patient presented with progressive cerebellar ataxia, and magnetic resonance imaging revealed slitlike supratentorial ventricles and a markedly enlarged fourth ventricle, which were compatible with the diagnosis of IFV. The surgical procedure described was performed under visualization through a styletlike slim optic fiberscope inserted into a ventricular catheter. The catheter, with the endoscope inside it, was passed through the foramen of Monro and then through the aqueduct to reach the enlarged fourth ventricle, where membranous occlusion of the foramen of Magendie was clearly visualized. The tip of the catheter was placed in the fastigium of the fourth ventricle. After the procedure, the size of the fourth ventricle was reduced and the patient's symptoms improved. Thus, it is concluded that endoscopic aqueductal stent placement is a simple and safe surgical procedure for treatment of IFV. PMID- 10839268 TI - Flushing in relation to a possible rise in intracranial pressure: documentation of an unusual clinical sign. Report of five cases. AB - This report documents clinical features in five children who developed transient reddening of the skin (epidermal flushing) in association with acute elevations in intracranial pressure (ICP). Four boys and one girl (ages 9-15 years) deteriorated acutely secondary to intracranial hypertension ranging from 30 to 80 mm Hg in the four documented cases. Two patients suffered from ventriculoperitoneal shunt malfunctions, one had diffuse cerebral edema secondary to traumatic brain injury, one was found to have pneumococcal meningitis and hydrocephalus, and one suffered an intraventricular hemorrhage and hydrocephalus intraoperatively. All patients were noted to have developed epidermal flushing involving either the upper chest, face, or arms during their period of neurological deterioration. The response was transient, typically lasting 5 to 15 minutes, and dissipated quickly. The flushing reaction is postulated to be a centrally mediated response to sudden elevations in ICP. Several potential mechanisms are discussed. Flushing has clinical importance because it may indicate significant elevations in ICP when it is associated with neurological deterioration. Because of its transient nature, the importance of epidermal flushing is often unrecognized; its presence confirms the need for urgent treatment. PMID- 10839269 TI - Selective transvenous liquid embolization of a Type 1 dural arteriovenous fistula at the junction of the transverse and sigmoid sinuses. Case report. AB - The authors describe the case of a 51-year-old man with a Type 1 dural arteriovenous fistula (AVF) located at the junction of the transverse and sigmoid sinuses. The dural AVF developed after the patient underwent a craniotomy for an acute extradural hematoma. The patient suffered pulsatile tinnitus 3 months after surgery. After several attempts at transarterial embolization (TAE), the venous channel located close to the skull fracture was accessed via a transfemoral transvenous approach and was embolized by administering a liquid nonadhesive agent. Successful embolization of the dural AVF was achieved both clinically and radiologically without causing considerable hemodynamic alterations. This procedure, either alone or combined with TAE, would seem to be an alternative treatment for dural AVFs in this location, without causing compromise of flow within the affected sinuses, when selective venous access is available. PMID- 10839270 TI - Modified stereotactic insertion of the Ommaya reservoir. Technical note. AB - Ommaya reservoirs are used primarily for the repeated injection of intrathecal chemotherapy for leptomeningeal metastasis from hematopoietic and solid malignancies. Insertion of this device in a relatively large nondisplaced ventricle is not a difficult task, but challenges arise when the ventricle is small and/or displaced. Different techniques have been developed to overcome this difficulty, most of which include the use of stereotactic frames. Further improvements would be beneficial. The technique described in this paper depends on a stereotactic frame; however, the modification proposed by the authors removes the arc system from the surgical field before the actual surgical procedure is begun. Removal of the arc improves access to the surgical field as well as preparation and draping of the surgical site and minimizes potential breaks in sterile technique, which ultimately reduces the incidence of infection. A twist-drill hole along the path of the chosen trajectory becomes an external guide for the ventricular catheter. The technique is easy, user friendly, and results in an unencumbered sterile field and reliable cannulation of small ventricles. A simple stereotactic technique for Ommaya reservoir insertion has been described. It should lower the chance of infection in this group of patients, most of whom have suppressed immune systems. PMID- 10839271 TI - A transparent sheath for endoscopic surgery and its application in surgical evacuation of spontaneous intracerebral hematomas. Technical note. AB - The authors advocate the use of a transparent sheath for guiding an endoscope, a simple and unique tool for endoscopic surgery, and describe preliminary results of its application in the evacuation of hypertensive intracerebral hematomas. This sheath is a 10-cm-long tube made of clear acrylic plastic, which greatly improves visualization of the surgical field through a 2.7-mm nonangled endoscope inserted within. Between April 1997 and December 1998, the authors performed endoscopic evacuation of intracerebral hematomas by using this sheath inserted into the patients' heads through a burr hole. In nine consecutive cases in which the hematoma was larger than 40 ml in volume, nearly complete evacuation (86 100%) of the lesion was achieved without complication. Excellent visualization of the border between the brain parenchyma and the hematoma facilitated accurate intraoperative orientation, and also allowed easy identification of the bleeding point. Thus, this combination of sheath and endoscope achieves both minimal invasiveness and the maximum extent of hematoma removal with secure hemostasis. This tool will reduce the inherent disadvantage of endoscopic procedures and may expand their application in other areas of neurosurgical management. PMID- 10839272 TI - Origin of the Drake fenestrated aneurysm clip. AB - The development of the Drake fenestrated aneurysm clip is a study in the history of ideas. This communication outlines the conception and solution of a surgical problem involved with the clipping of large basilar tip aneurysms. Dr. Charles G. Drake's ability to modify old ideas and experiment with new ones was instrumental to the conceptual idea of a fenestrated clip. Dr. Frank H. Mayfield and Mr. George Kees, Jr. played essential roles in bringing the idea to a reality. The development of the fenestrated clip has added substantially to the armamentarium of the aneurysm surgeon in dealing with large and complex aneurysms. PMID- 10839273 TI - Tuberculosis of the skull. Case illustration. PMID- 10839274 TI - Cyst of the transverse ligament. PMID- 10839275 TI - Nerve transfer. PMID- 10839276 TI - Iatrogenic pneumocephalus. PMID- 10839277 TI - Pathophysiology of syringomyelia. PMID- 10839278 TI - Pathophysiology of syringomyelia. PMID- 10839279 TI - Pathophysiology of syringomyelia. PMID- 10839280 TI - Endoscopic approach. PMID- 10839281 TI - Intracranial hemorrhage and ventriculoperitoneal shunts. PMID- 10839282 TI - Hypothermia and head injury. PMID- 10839283 TI - Blood-blister-like aneurysms. PMID- 10839284 TI - Intraoperative monitoring of electrooculograms. PMID- 10839285 TI - Magnetic source imaging. PMID- 10839286 TI - High-grade gliomas with chromosome 1p loss. PMID- 10839287 TI - Phase II study of second-line therapy with DTIC, BCNU, cisplatin and tamoxifen (Dartmouth regimen) chemotherapy in patients with malignant melanoma previously treated with dacarbazine. AB - This study assessed response rates to combination dacarbazine (DTIC), BCNU (carmustine), cisplatin and tamoxifen (DBPT) chemotherapy in patients with progressive metastatic melanoma previously treated with DTIC, as an evaluation of DBPT as a second-line regimen, and as an indirect comparison of DBPT with DTIC. Thirty-five consecutive patients received DBPT. The patients were divided into two groups. Group 1 comprised 17 patients with progressive disease (PD) on DTIC + tamoxifen therapy who were switched directly to DBPT. Group 2 comprised 18 patients not immediately switched to DBPT and included patients who had either a partial response (PR; one patient) or developed stable disease (SD; four patients) with DTIC, or received adjuvant DTIC (nine patients). All except four patients had received tamoxifen at the time of initial DTIC treatment. Median times since stopping DTIC were 22 days (range 20-41) and 285 days (range 50 1,240) in Groups 1 and 2 respectively. In Group 1, one patient developed SD for 5 months and the remainder had PD. In Group 2, there were two PRs, four patients with SD (4, 5, 6, and 6 months), and 11 with PD. These results indicate that the DBPT regimen is not of value in melanoma primarily refractory to DTIC. There were responses in patients not directly switched from DTIC to DBPT, suggesting combination therapy may be of value in a small subgroup of melanoma patients. PMID- 10839288 TI - Boron microlocalization in oral mucosal tissue: implications for boron neutron capture therapy. AB - Clinical studies of the treatment of glioma and cutaneous melanoma using boron neutron capture therapy (BNCT) are currently taking place in the USA, Europe and Japan. New BNCT clinical facilities are under construction in Finland, Sweden, England and California. The observation of transient acute effects in the oral mucosa of a number of glioma patients involved in the American clinical trials, suggests that radiation damage of the oral mucosa could be a potential complication in future BNCT clinical protocols, involving higher doses and larger irradiation field sizes. The present investigation is the first to use a high resolution surface analytical technique to relate the microdistribution of boron 10 (10B) in the oral mucosa to the biological effectiveness of the 10B(n,alpha)7Li neutron capture reaction in this tissue. The two boron delivery agents used clinically in Europe/Japan and the USA, borocaptate sodium (BSH) and p-boronophenylalanine (BPA), respectively, were evaluated using a rat ventral tongue model. 10B concentrations in various regions of the tongue mucosa were estimated using ion microscopy. In the epithelium, levels of 10B were appreciably lower after the administration of BSH than was the case after BPA. The epithelium:blood 10B partition ratios were 0.2:1 and 1:1 for BSH and BPA respectively. The 10B content of the lamina propria was higher than that measured in the epithelium for both BSH and BPA. The difference was most marked for BSH, where 10B levels were a factor of six higher in the lamina propria than in the epithelium. The concentration of 10B was also measured in blood vessel walls where relatively low levels of accumulation of BSH, as compared with BPA, was demonstrated in blood vessel endothelial cells and muscle. Vessel wall:blood 10B partition ratios were 0.3:1 and 0.9:1 for BSH and BPA respectively. Evaluation of tongue mucosal response (ulceration) to BNC irradiation indicated a considerably reduced radiation sensitivity using BSH as the boron delivery agent relative to BPA. The compound biological effectiveness (CBE) factor for BSH was estimated at 0.29 +/- 0.02. This compares with a previously published CBE factor for BPA of 4.87 +/- 0.16. It was concluded that variations in the microdistribution profile of 10B, using the two boron delivery agents, had a significant effect on the response of oral mucosa to BNC irradiation. From a clinical perspective, based on the findings of the present study, it is probable that potential radiation induced oral mucositis will be restricted to BNCT protocols involving BPA. However, a thorough high resolution analysis of 10B microdistribution in human oral mucosal tissue, using a technique such as ion microscopy, is a prerequisite for the use of experimentally derived CBE factors in clinical BNCT. PMID- 10839289 TI - Gemcitabine combined with continuous infusion 5-fluorouracil in advanced and symptomatic pancreatic cancer: a clinical benefit-oriented phase II study. AB - Gemcitabine and 5-fluorouracil are the only two compounds with reproducible activity against advanced pancreatic cancer (APC). We have evaluated a novel combination of gemcitabine and 5-fluorouracil on the clinical benefit response (CBR) end point. Eleven consecutive patients with symptomatic APC were entered in a two-stage phase II trial. Gemcitabine was administered by intravenous (i.v.) bolus injection at the dose of 1,000 mg m(-2) on days 1, 8, 15 and 5-fluorouracil 500 mg m(-2) was given by continuous i.v. infusion on days 1-5. Treatment was repeated every 28 days. A CBR was achieved in 7/11 patients. The mean time to loss of CBR was 26.5 weeks (range 14-18, median 22). Toxicity was mild and no APC patient experienced WHO grade 3 toxicity. The gemcitabine/5-fluorouracil combination is well tolerated and produces a symptomatic relief in the majority of APC patients. PMID- 10839291 TI - Reasons for accepting or declining to participate in randomized clinical trials for cancer therapy. AB - This paper reports on the reasons why patients agreed to or declined entry into randomized trials of cancer following discussions conducted by clinicians in both District General and University Hospitals. Two hundred and four patients completed a 16-item questionnaire following the consultation, of these 112 (55%) were women with breast cancer. Overall results showed that 147 (72.1%) patients accepted entry to a randomized clinical trial (RCT). The main reasons nominated for participating in a trial were that 'others will benefit' (23.1%) and 'trust in the doctor' (21.1%). One of the main reasons for declining trial entry was that patients were 'worried about randomization' (19.6%). There was a significantly higher acceptance rate for trials providing active treatment in every arm 98 (80.6%) compared with those trials with a no treatment arm 46 (60.5%), chi2 test P= 0.003. The study outlines a number of factors that appear to influence a patient's decision to accept or decline entry into an RCT of cancer therapy. An important factor is whether or not the trial offers active treatment in all arms of the study. Communication that promotes trust and confidence in the doctor is also a powerful motivating influence. PMID- 10839290 TI - Phase II study of the oxygen saturation curve left shifting agent BW12C in combination with the hypoxia activated drug mitomycin C in advanced colorectal cancer. AB - BW12C (5-[2-formyl-3-hydroxypenoxyl] pentanoic acid) stabilizes oxyhaemoglobin, causing a reversible left-shift of the oxygen saturation curve (OSC) and tissue hypoxia. The activity of mitomycin C (MMC) is enhanced by hypoxia. In this phase II study, 17 patients with metastatic colorectal cancer resistant to 5 fluorouracil (5-FU) received BW12C and MMC. BW12C was given as a bolus loading dose of 45 mg kg(-1) over 1 h, followed by a maintenance infusion of 4 mg kg(-1) h(-1) for 5 h. MMC 6 mg m(-2) was administered over 15 min immediately after the BW12C bolus. The 15 evaluable patients had progressive disease after a median of 2 (range 1-4) cycles of chemotherapy. Haemoglobin electrophoresis 3 and 5 h after the BW12C bolus dose showed a fast moving band consistent with the BW12C oxyhaemoglobin complex, accounting for approximately 50% of total haemoglobin. The predominant toxicities--nausea/vomiting and vein pain--were mild and did not exceed CTC grade 2. Liver 31P magnetic resonance spectroscopy of patients with hepatic metastases showed no changes consistent with tissue hypoxia. The principle of combining a hypoxically activated drug with an agent that increases tissue hypoxia is clinically feasible, producing an effect equivalent to reducing tumour oxygen delivery by at least 50%. However, BW12C in combination with MMC for 5-FU-resistant colorectal cancer is not an effective regimen. This could be related to drug resistance rather than a failure to enhance cytotoxicity. PMID- 10839292 TI - Survival benefit of chemotherapy in metastatic colorectal cancer: a meta-analysis of randomized controlled trials. AB - To estimate the magnitude of benefit of chemotherapy in prolonging survival for patients with metastatic colorectal cancer, a meta-analysis of randomized controlled trial was performed. A systematic search was performed to identify randomized trials comparing chemotherapy with observation or supportive care alone. Trials were assessed for quality of reporting, publication bias and heterogeneity. Relative risks for outcomes from published data were pooled using a random-effects model. Seven trials with 614 patients were included. All trials used fluoropyrimidine-based chemotherapy, through a variety of routes and schedules, including intravenous, intra-portal and hepatic arterial infusion. Compared with the 'no-chemotherapy' arm, chemotherapy significantly reduced 1 year mortality (risk ratio 0.69; 95% confidence interval (CI) 0.60-0.81, P < 0.00001). The mortality at 2 years was not significantly different (risk ratio 0.93; 95% CI 0.87-1.00, P = 0.053). Between-trial comparisons demonstrated benefit with a variety of routes and schedules. Chemotherapy significantly prolongs 1-year survival for patients with metastatic colorectal cancer, and should be offered to those with good performance status. PMID- 10839293 TI - Expression of transforming growth factor beta-1 in gastric cancer and in the gastric mucosa of first-degree relatives of patients with gastric cancer. AB - Transforming growth factors beta (TGF-beta) constitute a family of polypeptide growth factors that control cell growth, cell differentiation and migration, as well as the formation of the extracellular matrix. Recent analyses revealed the overexpression of TGF-beta1 in human gastric cancers and demonstrated increased cell proliferation in the stomach of patients with gastric cancer and their first degree relatives. Using human gastric tissues obtained from patients with gastric cancer (n = 19), biopsies from healthy first-degree relatives of gastric cancer patients (n = 18) and healthy individuals (n = 19), we analysed the expression of TGF-beta1 using the reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemistry. Fifteen of 19 patients with gastric cancer expressed TGF beta1 in the tumour. In 11 of these 15 cases TGF-beta1 mRNA was also detectable in the non-tumourous stomach. Interestingly, all but two individuals with a first degree relative diagnosed with gastric cancer exhibited TGF-beta1 expression in either the antrum or corpus biopsy or both. In contrast, only one of 19 individuals without a family history of gastric cancer expressed TGF-beta1 in the stomach (P< 0.0001). TGF-beta1 expression is detectable in a large proportion of gastric cancers and in the stomach of healthy first-degree relatives of gastric cancer patients. Since individuals without gastric cancers in their family express TGF-beta1 only in one of 19 cases, the induction of TGF-beta1 expression in first-degree relatives of patients with gastric cancer points to the presence of specific molecular alterations in a subgroup of individuals with an increased risk of developing gastric cancer that may precede the development of gastric cancers. PMID- 10839294 TI - Detailed deletion mapping of chromosome band 14q32 in human neuroblastoma defines a 1.1-Mb region of common allelic loss. AB - Neuroblastoma (NB) is a well-known malignant disease in infants, but its molecular mechanisms have not yet been fully elucidated. To investigate the genetic contribution of abnormalities on the long arm of chromosome 14 (14q) in NB, we analysed loss of heterozygosity (LOH) in 54 primary NB samples using 12 microsatellite markers on 14q32. Seventeen (31%) of 54 tumours showed LOH at one or more of the markers analysed, and the smallest common region of allelic loss was identified between D14S62 and D14S987. This region was estimated to be 1-cM long from the linkage map. Fluorescence in situ hybridization also confirmed the loss. There was no statistical correlation between LOH and any clinicopathologic features, including age, stage, amplification of MYCN and ploidy. We further constructed a contig spanning the lost region using bacterial artificial chromosome and estimated this region to be approximately 1.1-Mb by pulsed-field gel electrophoresis. Our results will contribute to cloning and characterizing the putative tumour-associated gene(s) in 14q32 in NB. PMID- 10839295 TI - Human tumour-associated cell adhesion protein MN/CA IX: identification of M75 epitope and of the region mediating cell adhesion. AB - MN/CA IX is a cell surface protein, strongly associated with several types of human carcinomas. It exerts activity of carbonic anhydrase and capacity of binding to cell surface receptors. In the present work, we used affinity purified MN/CA IX protein to demonstrate that the cells adhere to immobilized MN/CA IX and that the monoclonal antibody M75 abrogates cell attachment to MN/CA IX. Using synthetic oligopeptides, we identified M75 epitope and located it in the proteoglycan domain, which contains a sixfold tandem repeat of six amino acids GEEDLP. From phage display library of random heptapeptides we identified and chemically synthesized those which compete for the epitope with M75 and inhibit adhesion of cells to MN/CA IX. These heptapeptides might serve as lead compounds for drug design. PMID- 10839296 TI - Clinical usefulness of microsatellite instability for the prediction of gastric adenoma or adenocarcinoma in patients with chronic gastritis. AB - To assess a role of microsatellite instability (MSI) in the development of gastric adenocarcinoma or adenoma from chronic gastritis, we analysed mutations of five microsatellite loci in gastritis, adenoma and adenocarcinoma retrospectively. Gastric mucosa was biopsied from the same area in each patient at different periods and examined for MSI. Only one of 55 patients with chronic gastritis revealed MSI-H phenotype and the other 54 patients showed microsatellite stable (MSS) phenotypes. In six of 17 patients with gastric adenoma or well-differentiated adenocarcinoma, MSI-positive phenotypes were demonstrated. Interestingly, all of six patients showing MSI, including three high-level MSI (MSI-H) cases and three low-level (MSH-L) cases, had already revealed MSI at the stage of chronic gastritis. In two of three MSI-H cases, the identical MSI patterns had been observed at the stage of gastritis 1.5-7 years before the final diagnosis of adenocarcinoma. The adjacent gastritis mucosa within 10 mm from the carcinoma demonstrated MSI as well. MSI was not found in any of 35 patients with Helicobacter pylori infection, but found in one of 30 patients without infection. Moreover, two of three cases of gastric adenoma or well-differentiated adenocarcinoma with MSI-H at the stage of chronic gastritis showed no evidence of Helicobacter pylori infection throughout the observation periods. These results indicate that MSI in biopsy specimens at the stage of chronic gastritis may predict the risk of the progression to adenoma and well differentiated adenocarcinoma, and that Helicobacter pylori infection itself may not induce MSI directly in the gastric mucosa. PMID- 10839297 TI - Up-regulation of telomerase activity in human pancreatic cancer cells after exposure to etoposide. AB - Telomerase plays a critical role in the development of cellular immortality and oncogenesis. Activation of telomerase occurs in a majority of human malignant tumours, and the relation between telomerase and vulnerability to drug-mediated apoptosis remains unclear. In this study, we demonstrate, for the first time, up regulation of telomerase activity in human pancreatic cancer cells treated with etoposide, a topoisomerase II inhibitor. Exposure of MIA PaCa-2 cells to etoposide at various concentrations (1-30 microM) resulted in two- to threefold increases in telomerase activity. Up-regulation was detectable 24 h after drug exposure and was accompanied by enhanced expression of mRNA of the human telomerase reverse transcriptase. Telomerase activation was also observed in AsPC 1 and PANC-1 cells but not in KP-3 and KP-1N cells. Furthermore, we found a negative correlation between increased telomerase activity and the percentage of dead cells after etoposide treatment. These findings suggest the existence of an anti-apoptotic pathway through which telomerase is up-regulated in response to DNA damage. This telomerase activation pathway may be one of the mechanisms responsible for the development of etoposide resistance in certain pancreatic cancer cells. PMID- 10839298 TI - Activation of SAPK/JNK by camptothecin sensitizes androgen-independent prostate cancer cells to Fas-induced apoptosis. AB - We have previously shown that the androgen-independent prostate cancer cells DU145, despite expressing Fas and FasL, were resistant to anti-Fas-induced apoptosis, and that this resistance could be overcome by pretreating the cells with sublethal doses of camptothecin. Here, we provide evidence that SAPK/JNK activity is required for camptothecin sensitization to anti-Fas-induced apoptosis. Camptothecin, but not Fas ligation, was shown to activate SAPK/JNK in a time-dependent manner, and to induce c-Jun expression. The effects were more prominent in cells treated with both camptothecin and anti-Fas. The expression levels of MKP-1, a phosphatase which regulates SAPK/JNK and which has been implicated in prostate cancer resistance to apoptosis, remained unchanged. Inhibition of caspases had no effect on the SAPK/JNK activation, suggesting that this activation is an upstream event in the Fas-signalling pathway, and is independent of caspase activity. Antisense oligonucleotides targeted to JNK1 and JNK2 reversed the effect of camptothecin. These results suggest that stress kinase activation can significantly influence the fate of androgen-independent prostate cancer cells following Fas receptor ligation. PMID- 10839300 TI - Development and characterization of a tamoxifen-resistant breast carcinoma xenograft. AB - A human tamoxifen-resistant mammary carcinoma, MaCa 3366/TAM, originating from a sensitive parental xenograft 3366 was successfully established by treatment of tumour-bearing nude mice with 1-50 mg kg(-1) tamoxifen for 3 years during routine passaging. Both tumours did not differ significantly in OR- and PR-positivity, however, when compared with the sensitive tumour line, the mean OR content of the TAM-resistant subline is slightly lower. An OR-upregulation following withdrawal of oestradiol treatment was observed in the parental tumours but not in the resistant xenografts. Following long-term treatment with tamoxifen, the histological pattern of the breast carcinoma changed. The more differentiated structures being apparent after treatment with 17beta-oestradiol in the original 3366 tumour were not induced in the resistant line. Tamoxifen failed to induce a tumour growth inhibition in comparison to the tamoxifen-sensitive line. The pure anti-oestrogen, ICI 182 780, revealed cross-resistance. Sequence analysis of the hormone-binding domain of the OR of both lines showed no differences, suggesting that either mutations in other regions of the OR are involved in the TAM resistance phenotype or that mechanisms outside of this protein induced this phenotype. Oestrogen and anti-oestrogen regulate pS2 and cathepsin D expression in 3366 tumours as in the human breast cancer cell line MCF-7. The resistant 3366/TAM tumours have lost this regulation. The established breast cancer xenografts 3366 and 3366/TAM offer the possibility of investigating mechanisms of anti-oestrogen resistance in an in vivo situation. They can be used to test novel approaches to prevent, or to overcome, this resistance in a clinically related manner. PMID- 10839299 TI - Nitric oxide production by tumour tissue: impact on the response to photodynamic therapy. AB - The role of nitric oxide (NO) in the response to Photofrin-based photodynamic therapy (PDT) was investigated using mouse tumour models characterized by either relatively high or low endogenous NO production (RIF and SCCVII vs EMT6 and FsaR, respectively). The NO synthase inhibitors Nomega-nitro-L-arginine (L-NNA) or Nomega-nitro-L-arginine methyl ester (L-NAME), administered to mice immediately after PDT light treatment of subcutaneously growing tumours, markedly enhanced the cure rate of RIF and SCCVII models, but produced no obvious benefit with the EMT6 and FsaR models. Laser Doppler flowmetry measurement revealed that both L NNA and L-NAME strongly inhibit blood flow in RIF and SCCVII tumours, but not in EMT6 and FsaR tumours. When injected intravenously immediately after PDT light treatment, L-NAME dramatically augmented the decrease in blood flow in SCCVII tumours induced by PDT. The pattern of blood flow alterations in tumours following PDT indicates that, even with curative doses, regular circulation may be restored in some vessels after episodes of partial or complete obstruction. Such conditions are conducive to the induction of ischaemia-reperfusion injury, which is instigated by the formation of superoxide radical. The administration of superoxide dismutase immediately after PDT resulted in a decrease in tumour cure rates, thus confirming the involvement of superoxide in the anti-tumour effect. The results of this study demonstrate that NO participates in the events associated with PDT-mediated tumour destruction, particularly in the vascular response that is of critical importance for the curative outcome of this therapy. The level of endogenous production of NO in tumours appears to be one of the determinants of sensitivity to PDT. PMID- 10839301 TI - Deficient activation of CD95 (APO-1/Fas)-mediated apoptosis: a potential factor of multidrug resistance in human renal cell carcinoma. AB - The pronounced resistance of human renal cell carcinoma (RCC) to anticancer induced apoptosis has primarily been related to the expression of P-glycoprotein and effective drug detoxification mechanisms. Because the CD95 system has recently been identified as a key mediator of anticancer drug-induced apoptosis, we analysed the contribution of the CD95 system to chemotherapy-induced apoptosis in four newly established RCC cell lines. Here, we demonstrate that all RCC cell lines expressed CD95-receptor and -ligand. Exposure to agonistic anti-CD95 antibodies resulted in induction of apoptosis and significant (P < 0.05) reduction of cell number in three out of four cell lines, indicating that the essential components for CD95-mediated apoptosis were present and functionally intact in the majority of these RCC cell lines. Moreover, treatment of cultures with bleomycin or topotecan, a novel topoisomerase I inhibitor with little substrate affinity for P-glycoprotein, led to induction of apoptosis and significant (P < 0.05) dose-dependent reduction of cell number in all RCC cell lines. Both anticancer drugs also induced upregulation of CD95 ligand expression in all cell lines. Additionally, augmentation of CD95 receptor expression was found in three RCC cell lines, including one p53-mutated cell line, whereas another p53-mutated cell line showed no or only a weak CD95 receptor upregulation after exposure to topotecan or bleomycin, respectively. Despite this upregulation of CD95 receptor and ligand, antagonistic antibodies directed against CD95 receptors or ligands could not inhibit induction of apoptosis by topotecan and bleomycin in any cell line. Thus, although a functionally intact CD95 signalling cascade is present in most RCC cell lines, the anticancer drugs topotecan and bleomycin that induce upregulation of CD95 receptor and ligand fail to effectively activate CD95-mediated apoptosis. This deficient activation of CD95 mediated apoptosis might be an important additional factor for the multidrug resistance phenotype of human RCCs. PMID- 10839302 TI - Menopause and colorectal cancer. AB - Post-menopausal women who have never used hormone replacement therapy have a higher risk of colon, but not rectal, cancer than do premenopausal women of the same age, socio-cultural class and dietary habits. Such risk increase seems to last about 10 years and to be restricted to lean women, a group who have lower levels of oestradiol after ovarian function ceases after menopause. PMID- 10839303 TI - Rural factors and survival from cancer: analysis of Scottish cancer registrations. AB - In this survival study 63,976 patients diagnosed with one of six common cancers in Scotland were followed up. Increasing distance from a cancer centre was associated with less chance of diagnosis before death for stomach, breast and colorectal cancers and poorer survival after diagnosis for prostate and lung cancers. PMID- 10839304 TI - Ethnic differences in post-menopausal plasma oestrogen levels: high oestrone levels in Japanese-American women despite low weight. AB - Breast cancer incidence in Japanese-American women is approaching that of US Whites. We investigated whether this shift is paralleled by similar post menopausal plasma hormone levels in the two ethnic groups. We also included African-American and Latina women to further our understanding of possible ethnic differences in oestrogen metabolism. We measured androstenedione (A), oestrone (E1) and oestradiol (E2) in 30 Japanese-American, 39 non-Latina White ('White'), 66 African-American and 58 Latina women. The (age-adjusted) geometric mean E1 levels were 34 pg ml(-1) in Japanese-Americans, 28 pg ml(-1) in Whites, 35 pg ml( 1) in African-Americans and 31 pg ml(-1) in Latinas. After adjustment for body mass index, Japanese-Americans had the highest mean E1 value of all groups and this was statistically significantly greater than the value for Whites (P(t-test) = 0.05). The geometric mean A concentrations were also highest in Japanese Americans. There was little ethnic difference in E2 levels. In conclusion, post menopausal plasma oestrogen levels in Japanese-American women are at least as high as those in Whites. PMID- 10839305 TI - Risk factors for leukoplakia and malignant transformation to oral carcinoma: a leukoplakia cohort in Taiwan. AB - The effects of betel nut chewing, smoking and alcohol on the occurrence of leukoplakia and its malignant transformation to oral carcinoma were quantified in a leukoplakia cohort (n = 435) from one medical centre between 1988 and 1998 in Taiwan. Sixty oral carcinomas were ascertained in this cohort. A case-control study within the leukoplakia cohort was used to study, risk factors. Using the Weibull survival model, the incidence of malignant transformation of leukoplakia was shown to increase with follow-up years. After adjustment for other relevant risk factors, betel nut chewing (adjusted odds ratio (OR) = 4.59; 95% confidence interval (CI) 1.25-16.86) remained a significant risk factor for malignant transformation. Results from the case-control study showed that the adjusted odds ratios for betel nut chewing and smoking on the occurrence of leukoplakia were 17.43 (95% CI 1.94-156.27) and 3.22 (95% CI 1.06-9.78), respectively. Similar findings were observed when daily frequency and duration were taken into account. This implies that cessation of smoking may reduce by 36% leukoplakia cases, while elimination of betel nuts may prevent 62% of leukoplakia and 26% of malignant transformation to oral carcinoma in the underlying population. PMID- 10839306 TI - Retinoblastoma incidence and sunlight exposure. AB - To evaluate positive findings from an earlier report, we studied the relation between retinoblastoma incidence and ultraviolet (UV-B) radiation levels in the Surveillance, Epidemiology, and End Results (SEER) programme areas of the USA using weighted regression, as well as in international data after adjusting for race, economic development, and climate. The association was not statistically significant within the USA (P > 0.20). At an international level, the relation was significant overall and after adjusting for economic development, but it was not significant after adjusting for race and tropical climate, suggesting that environmental factors other than UV-B may be responsible for the geographic patterns of retinoblastoma. PMID- 10839307 TI - Effects of soy foods on ovarian function in premenopausal women. AB - It has been proposed that the high intake of soy foods among Asians may partly explain their lower rates of breast cancer, perhaps by lowering endogenous oestrogen levels, although this has been inadequately studied. Twenty healthy cycling premenopausal women (ten Asians and ten non-Asians) participated in a 7 month soy intervention study which was designed to investigate the effect of supplementation on ovarian function. Asian soy foods (tofu, soymilk, green soybean peas) in the amount of approximately 32 mg of isoflavones per day were added to the women's diets for three menstrual cycles. The women's baseline (two cycles) serum hormone levels were compared to levels during soy intervention (three cycles) and levels after intervention (two cycles). During the entire study period, subjects provided almost daily overnight urine samples and blood specimens during specified days of their menstrual cycles. The day of urinary luteinizing hormone (LH) peak was used as a marker for the day of ovulation. Knowledge of day of ovulation allowed comparison of hormone measurements at baseline to those obtained during intervention and recovery cycles with standardization of day of cycle. Soy intervention was associated with a statistically significant reduction in serum luteal oestradiol level (-9.3%, P < 0.05), but there were no significant changes in follicular phase oestradiol, follicular or luteal phase progesterone, sex hormone-binding globulin or menstrual cycle length. This significant reduction in luteal phase oestradiol was, however, observed only among Asian (-17.4%) but not among non-Asian (-1.2%) participants; urinary excretion of isoflavones was higher among Asians than non Asians (29.2 vs 17.1 micromol day(-1), P= 0.16) during the intervention period. Thus, supplementation using traditional soy foods reduced serum oestradiol levels among Asian participants in this study. Differences in the type of soy products (i.e. traditional soy foods versus soy protein products), amount of isoflavones, and race/ethnicity of participants may have contributed to the divergent results. Larger soy intervention studies designed specifically to include participants of different race/ethnicities and using both traditional soy foods and soy protein products providing comparable doses of isoflavones are needed to definitively determine the effect of soy on ovarian function. PMID- 10839308 TI - Mortality from cutaneous melanoma: evidence for contrasting trends between populations. AB - In recent years several reports have been published concerning trends in melanoma mortality in different countries, some of which have indicated that rates are beginning to fall. Many of these reports, however, have been based on small populations and have used different forms of statistical analysis. Our objective was to analyse systematically to what degree the epidemic of melanoma mortality had evolved similarly in different populations and whether there were any divergent trends that might increase our understanding. Instead of using all available data, we focused on countries with a minimum time series of 30 years and a minimum of 100 deaths annually in at least one sex from melanoma. We first inspected sex-specific age-standardized mortality rates and then performed age period-cohort modelling. We found that the increase in mortality observed after 1950 was more pronounced in the age group 60-79. Statistical modelling showed a general increase in mortality rates in generations born after the turn of the century. Downturns in mortality, essentially in women and starting with generations born just before World War II, were found in Australia (where the earliest decreases were noted), the Nordic countries and the USA. Small decreases in rates in more recent generations were found in the UK and Canada. However, in France, Italy and Czechoslovakia, mortality rates were seen to be still increasing in recent cohorts. Our analysis suggests that populations are at different places on the melanoma mortality epidemic curve. The three trend patterns we observed are in agreement with time differences between populations with respect to the promotion of sun protection and the surveillance of pigmented skin lesions. PMID- 10839309 TI - Alcohol consumption and cigarette smoking in relation to high frequency of p53 protein accumulation in oesophageal squamous cell carcinoma in the Japanese. AB - We investigated levels of p53 protein expression in Japanese patients with oesophageal squamous cell carcinoma. A significantly larger proportion of heavy alcohol drinkers and cigarette smokers was evident in the p53-positive group. The combination of drinking and smoking was associated with a high frequency of p53 protein accumulation. PMID- 10839310 TI - Serum vascular endothelial growth factor load and interleukin-6 in cancer patients. PMID- 10839313 TI - The effect of exposure to a commercial 2,4-D herbicide formulation during gestation on urethan-induced lung adenoma formation in CD-1 mice. AB - Female CD-1 mice were exposed to a commercial amine formulation of 2,4 dichlorophenoxyacetic acid (2,4-D) on days 6-16 of gestation in drinking water at concentrations ranging from 0 to 1.0% of the formulated product, equivalent to approximately 0-650 mg/kg/d expressed as the amine derivative. The effect of 2,4 D on urethan-induced pulmonary adenoma formation was evaluated in female offspring 19 w after birth. Urethan-induced sleeping times observed following ip injection of 1.5 mg urethan/g bw 7 w after birth were not altered by 2,4-D (p = 0.10), indicating that 2,4-D did not affect the rate of urethan elimination. 2,4 D exposure did not affect the number of tumors produced (p = 0.58), but did reduce the mean tumor diameter in the highest dose group (p < 0.01). This minor antineoplastic activity of 2,4-D may be related, in part, to inhibitory effects of 2,4-D on various enzymatic or metabolic pathways, essential for cellular growth and tissue development. Since exposure to 2,4-D during pregnancy had little impact of tumor production, it is unlikely that persistent alteration to developing immune cells involved in the cell-mediated immunosurveillance mechanisms occurred. The subtle alteration in tumor size and the mild impairment of growth in the offspring were observed almost exclusively in the highest treatment group. Since this level of exposure is well in excess of those associated with normal application of 2,4-D, the hazard to non-target mammalian populations appears minimal. PMID- 10839312 TI - Beyond speech lateralization: a review of the variability, reliability, and validity of the intracarotid amobarbital procedure and its nonlanguage uses in epilepsy surgery candidates. AB - While the intracarotid amobarbital procedure (IAP) was originally utilized to lateralize speech functions as an aid in the surgical treatment of epilepsy, additional uses for the IAP have emerged including: (1) the use of the IAP to predict post-surgical memory changes, including both global amnesia and smaller, yet significant, material-specific memory deficits; (2) the use of the IAP to provide confirmatory evidence of lateralization of seizure focus; and (3) the use of the IAP to predict post-surgical relief from seizures. While the literature on the IAP is extensive and growing, its utility is marred by the wide variability associated with the procedure itself from epilepsy center to center. This variability renders comparisons among IAP studies problematic and conclusions about IAP efficacy difficult. The variability associated with the amobarbital procedures, as well as the reliability and the validity of the IAP in its nonlanguage uses is reviewed here. A special emphasis is devoted to research conducted in the last decade. Also discussed is the future of the IAP including anticipated research directions. PMID- 10839311 TI - Does smoking by pregnant women influence IQ, birth weight, and developmental disabilities in their infants? A methodological review and multivariate analysis. AB - Neuropsychologists are asked frequently to address the issue of the cause of a variety of central nervous system problems that may affect higher cortical function. One such issue is the relationship of maternal smoking to adverse reproductive outcomes involving neocortical insult including mental retardation, learning disabilities, attention-deficit hyperactivity disorder, and other insults that may be related to prolonged hypoxic states in utero. The instant paper develops the issue of causation as a scientific inquiry, reviews several traditional, applicable models, and critiques these models. An additional model of motility is proposed and discussed. The issue of the relationship of maternal smoking to adverse reproductive outcomes is then addressed from a review perspective along with new empirical analyses, the latter demonstrating that researchers tend to draw causal conclusions independent of whether the respective design of their studies would support conclusions about the causation of an event. Causal conclusions in the absence of causal designs have often lead to incomplete and incorrect conclusions. It is necessary to match conclusions not only to the outcomes of a research project but also to its design and accompanying limitations. PMID- 10839314 TI - Effects of pearl millet (Pennisetum typhoides), and fermented and processed fermented millet on Nubian goats. AB - The effects of Pearl millet, fermented millet, or processed fermented millet were investigated in 15 goats assigned as untreated controls, Pearl millet-fed (166.6g/animal/d), fermented millet-fed (166.6 g/animal/d or 83.3g/animal/d), and processed fermented millet-fed (166.6g/ animal/d). Pearl millet caused goiter within 72-108 d, and changes in thyroid follicles were correlated with clinical manifestations and alterations in serum and tissue iodine and selenium concentrations. Neither fermented nor processed fermented millet produced goiter, but their toxicity was characterized by ruminal bloat, dyspnea, diarrhea, ataxia, and enterohepatonephrotoxicity with death within 5-15 d. Organ lesions correlated with changes in serum enzymes and other serum constituents, hematology, and tissue and serum iodine and selenium concentrations. PMID- 10839315 TI - Plant poisonings from herbal medication admitted to a Tunisian toxicologic intensive care unit, 1983-1998. AB - To collect data on Tunisian herbal toxicity we analysed 56 medical records of patients admitted to the toxicological intensive care unit from 1983 to 1998 following the ingestion of plants. The sex ratio of patients was 1:2, the mean age of patients was 26y (2 to 75y) and II species of plants were involved with Atractylis gummifera (32%), Datura stramonium (25%), Ricinus communis (9%), Nerium oleander (7%) and Peganum harmala (7%) most commonly infested. Poisonings involved neurological (91%), gastrointestinal (73%) and cardiovascular systems (18%). Treatment was mainly symptomatic. The only lethal cases of liver failure involved 16 Atractylis gummifera poisonings. PMID- 10839316 TI - Isolation of steroidal sapogenins implicated in experimentally induced cholangiopathy of sheep grazing Brachiaria decumbens in Brazil. AB - As part of an experimental study, crystal-associated cholangiopathy was induced in 9 sheep by grazing pure pastures of Brachiaria decumbens in Brazil. One of these sheep showed characteristic lesions of photosensitization. The analysis of the B decumbens samples by acidic hydrolysis followed by TLC and infrared spectrum revealed diosgenin as the principal sapogenin present in the plant. In the rumen contents samples from the B decumbens-grazing group were identified by TLC, 1H and 13C NMR and EIMS as epismilagenin, episarsasapogenin, and a mixture of smilagenin and sarsasapogenin. In the bile samples from the B decumbens grazing group, TLC analysis demonstrated 2 compounds similar to epismilagenin and episarsasapogenin. However, by this same method, those compounds were not observed in the rumen contents and bile from 2 sheep which served as control animals. The P chartarum spore counts remained very low during the experimental period. PMID- 10839317 TI - Acute acetaminophen toxicity in transgenic mice with elevated hepatic glutathione. AB - Previous studies demonstrated that elevation of hepatic glutathione (GSH) concentrations protect against acetaminophen (APAP) hepatotoxicity in mice. Employing transgenic mice overexpressing glutathione synthetase, this study was conducted to determine if sustained elevation of hepatic GSH concentrations could ameliorate or prevent APAP toxicity. International Cancer Research transgenic mouse males and matched (ie same strain, sex, and age) control nontransgenic mice were pretreated ip with GSH synthetase substrate gamma-glutamylcysteinyl ethyl ester (gamma-GCE) or with saline. After a 16-h fast, mice received a single dose of 500 mg APAP/kg bw in saline ip and were sacrificed 4 h later. Other mice similarly pretreated were killed without APAP challenge. The elevated GSH concentrations in transgenic mice livers did not lessen APAP hepatotoxicity. Instead higher degrees of hepatotoxicity and nephrotoxicity were observed in transgenic mice than in controls as indicated by higher serum alanine aminotransferase activity and more severe histopathological lesions in transgenic mice livers and kidneys. Pretreatment with gamma-GCE did not affect either initial or post-APAP treatment tissue GSH concentrations or observed degrees of toxicity. Detection of a higher level of serum APAP in transgenic mice and the histopathological lesions found in transgenic mice kidneys together with no observable nephrotoxicity in control mice indicated early kidney damage in transgenic mice. Our findings suggest that high levels of GSH-APAP conjugates resulting from increased GSH concentrations in the livers of transgenic mice caused rapid kidney damage. Compromised excretory ability may have caused retention of APAP, which, in effect, elicited higher hepatotoxicity than that observed in nontransgenic mice. PMID- 10839318 TI - Detecting Taxus poisoning in horses using liquid chromatography/mass spectrometry. AB - A method is described for the analysis of taxine alkaloids by liquid chromatography/mass spectrometry. It is applicable to the detection of taxine alkaloids in the stomach contents of horses in which Taxus poisoning is suspected. Analysis of a leaf extract of Taxus baccata revealed unreported alkaloids of the same relative molecular mass as taxine B and isotaxine B. PMID- 10839319 TI - Neuronal vacuolation of the trigeminal nuclei in goats caused by ingestion of Prosopis juliflora pods (mesquite beans). AB - Three groups of 6 goats each were fed a ration containing 30, 60, or 90%, on a dry matter base, of Prosopis juliflora pods. A control group of 4 goats ingested only the basic ration. Two hundred and ten days after the start of the experiment 3 goats that ingested 60% pods in and 4 that ingested 90% had mandibular tremors, mainly during chewing. All animals were killed after 270 d of ingestion. No gross lesions were observed. Histologic lesions were characterized by fine vacuolation of the pericaryon of neurons from the trigeminal nuclei. Occasionally neurons of the oculomotor nuclei were also affected. Wallerian degeneration was occasionally observed in the mandibular and trigeminal nerves. Denervation atrophy of the masseter, temporal, hypoglossus, genioglossus, styloglossus, medial pterygoid and lateral pterygoid muscles was seen. The clinical signs from feeding the P juliflora pods were caused by a selective toxicity to neurons of some cranial nerve nuclei. PMID- 10839320 TI - Changes in hepatic and renal glutathione-dependent enzyme activities in rabbits and lambs subchronically treated with triphenyltin acetate. AB - To gain insight into the biochemical mechanisms of organotin toxicity, the effects of oral subchronic exposure (70 d) to triphenyltin acetate (TPTA) on hepatic and renal enzymes involved in glutathione metabolism were investigated in rabbits and lambs. Rabbits were offered a diet fortified with 15, 75 or 150 ppm TPTA, whereas lambs were daily given 1 or 7.5 mg/kg TPTA On the whole, rabbits were more susceptible than lambs and in both species hepatic enzymes were affected to a greater extent than renal enzymes. In rabbit liver, glutathione S transferase activity toward 1,2-dichloro-4-nitrobenzene (DCNB) was enhanced at 15 ppm and depressed at 150 ppm TPTA, whereas selenium-dependent glutathione peroxidase (Se-GPX) decreased in a dose-related manner; glyoxalase II (GII) activity increased to the same extent at 15 or 75 ppm TPTA but was unaffected at 150 ppm TPTA. For renal enzyme activities in rabbits, only GPX activity was significantly inhibited at 150 ppm TPTA. The only statistically significant changes in lambs were a fall in both hepatic GST accepting DCNB as substrate at 7.5 mg/kg and Se-GPX at 1 or 7.5 mg/kg TPTA, and an increase in renal GII activity at 7.5 mg/kg TPTA. These results suggest that depression of important antioxidant enzymes such as GST and GPX are part of the complex mechanism of organotin toxicity. PMID- 10839321 TI - Botulism-like syndrome after injections of botulinum toxin. AB - Botulinum type A toxin (BTA) is an orphan drug used to treat several disorders of muscle spasticity. We report the first known case of systemic botulism-like syndrome induced by BTA therapy which resulted in respiratory arrest. Clinicians should be aware that systemic effects may occur with localized BTA therapy and may be life-threatening. PMID- 10839322 TI - Marked elevation of urinary zinc levels and pleural-friction rub in metal fume fever. AB - Zinc fume is a well-recognized cause of metal fume fever, characterized by acute or subacute symptoms of respiratory tract inflammation, myalgias and fever (similar to influenza) associated with a variety of metal oxide fumes. A welder of galvanized steel developed metal fume fever, a pleural friction rub and markedly elevated urinary zinc excretion despite work-site surveillance data indicating exposures close to the Occupational Safety and Health Administration, Permissible Exposure Limit. This patient suggests that exposure data may be misleading. This case had the highest urine zinc levels reported in the literature and a previously unreported coexistent pleural friction rub. The symptoms resolved with administrative and engineering controls to decrease exposure. PMID- 10839323 TI - Mushroom toxicity in a horse with meningioangiomatosis. AB - We describe a fatal case of mushroom intoxication in an 18-y-o horse presumably due to Amanita verna. Horses are normally regarded as too fastidious to eat the ill-flavored toadstools. In this case, the horse had a rare benign brain tumor, meningioangiomatosis, which may have altered the horse's normal eating behavior resulting in consumption of the mushrooms. PMID- 10839324 TI - Association of sympathomimetic drugs with malformations. AB - We observed 2 cases of severe limb defects in infants following the use of sympathomimetic drugs during pregnancy. The mother of I had taken large doses of Primatene (ephedrine, theophylline, phenobarbital) as tablets and mist throughout pregnancy. The infant was born with oligoectrosyndactyly. In the other infant, maternal ingestion of sympathomimetic drugs including Triaminic (pseudoephedrine, phenylephrine, phenylpropanolamine) was associated with distal limb defects. Experimental studies in pregnant rabbits using Primatene in both low and high dosage resulted in limb reduction defects and other malformations in a significant number of the offspring compared with controls. Limb defects in the offspring of chicks following exposure to sympathomimetic drugs had previously been observed. We suggest that these agents may be teratogenic in the human and should be used with great caution or avoided during pregnancy. PMID- 10839325 TI - Congestive heart failure and toxic digoxin levels: role of cholestyramine. AB - We report 2 patients with congestive heart failure and concomitant toxic serum digoxin levels in whom cholestyramine administration increased clearance of digoxin and decreased serum digoxin elimination half-lives. Cholestyramine's gastrointestinal binding of enterohepatically-recycled and enterally-secreted digoxin enhanced elimination, allowing a more rapid return to therapeutic drug levels. PMID- 10839326 TI - Environmental factors associated with blood lead levels in Venezuelan children. AB - A preliminary study explored the relative contribution of residential sources of lead exposure on mentally challenged children who attend "special education" institutions (GI) compared to a group of age and sex matched school children (G2). We captured descriptive information and analyzed demographic variables, personal and household information, medical effects, environmental exposure factors, and children habits. Home paint, dust, soil, and water sampling was conducted and blood lead (BPb) levels determined. Eighteen G1 and 20 G2 children were studied. The mean G1 BPb was 16.9 +/- 7.9 microg/dl and was significantly higher than that in G2. Fifty percent of G1 children had PbB >20 microg/dl and 72.2% were >10 microg/dl. Low muscular strength, decreased osteotendinose reflexes, fine and gross motricity, deficient equilibrium, and hipotonic muscular tone coincided with >18 microg/dl BPb levels. In 61.1% of G1 homes paint lead levels were higher than permissible levels and 33.3% had dust lead exceeding that level. The high BPb levels in G1 probably resulted from ingestion of household paint, dust, and soil via "hand-to-mouth" activity. Environmental exposure to lead can be an important source of lead intake by infants and children and could affect neurological development. This study provides new insights currently unavailable for these children in Venezuela. PMID- 10839327 TI - Dealing with "risk" at the millennium. PMID- 10839328 TI - Fatal privet toxicosis in Tennessee cows...continued. PMID- 10839329 TI - The plant is Urginea maritima (L) Baker. PMID- 10839330 TI - Decisional capacity for informed consent in schizophrenia research. AB - BACKGROUND: The adequacy of subjects' informed consent to research is the focus of an important public and professional debate. The potential impairment of decisional capacity in persons with schizophrenia is central to the discussions. This study ascertains the decisional capacity for informed consent in schizophrenic research subjects, to determine if reduced capacity relates to specific aspects of psychopathologic features and to test the hypothesis that reduced capacity can be remediated with an educational informed consent process. METHODS: Decisional capacity was assessed for 30 research subjects with schizophrenia and 24 nonill (normal) comparison subjects. Measures of psychopathologic features and cognition were obtained for the subjects with schizophrenia. Subjects who performed poorly on the decisional capacity measure received an educational intervention designed to improve their ability to provide informed consent and were then retested. RESULTS: The patient group did not perform as well as the controls on initial decisional capacity assessment. Poor performance was modestly related to the extent of symptoms but robustly related to cognitive impairments. Following the educational intervention, the performance of subjects with schizophrenia was equal to that of the nonill comparison group. CONCLUSIONS: Many persons with schizophrenia may be challenged by the cognitive demands of an informed consent process for research participation. In many cases, their reduced capacity can be compensated by a more intensive educational intervention as part of the informed consent process. PMID- 10839331 TI - Evidence-based ethics and informed consent in mental illness research. PMID- 10839332 TI - Marijuana and medicine: assessing the science base: a summary of the 1999 Institute of Medicine report. AB - In response to public pressure to allow the medical use of marijuana, the Office of National Drug Control Policy, Washington, DC, funded a study by the Institute of Medicine evaluating the scientific evidence for benefits and risks of using marijuana as a medicine. The report used scientific reviews, public hearings, and reports from other agencies, and was evaluated by knowledgeable advisors and reviewers. It called for heavier investment in research on the biology of cannabinoid systems, careful clinical studies of cannabinoids in clinical syndromes, analysis of cannabinoids' psychological effects on symptoms, and evaluations of the health consequences of heavy marijuana use; recommends against the use of smoked marijuana in medicine and for the development of a medical cannabinoid inhaler; and recommends that compassionate use of marijuana be considered under carefully reviewed protocols. Finally, the report evaluates the abuse potential, tolerance, withdrawal, and gateway risks of medical use of cannabinoid drugs. PMID- 10839333 TI - A positron emission tomography study of quetiapine in schizophrenia: a preliminary finding of an antipsychotic effect with only transiently high dopamine D2 receptor occupancy. AB - BACKGROUND: Quetiapine is a new atypical antipsychotic medication. As such, relatively little has been published regarding its in vivo effects at the dopamine type 2 (D2) and serotonin type 2a (5-HT2a) receptor systems. The following study was undertaken to explore these effects across the clinical dose range and relate this information to its clinical profile. METHODS: Twelve patients with schizophrenia were randomly assigned to doses of 150 to 600 mg/d (n=3, at 150, 300, 450, and 600 mg/d) of quetiapine. After 3 weeks of treatment, D2 and 5-HT2a occupancy were measured using positron emission tomography (PET) imaging, 12 to 14 hours after the last dose. Clinical efficacy and adverse effect ratings were obtained at baseline, at the time of PET scanning, and at 12 weeks. Two additional patients were included to examine the effects of the drug 2 to 3 hours after last dose. RESULTS: Quetiapine was an effective antipsychotic and improved the extrapyramidal symptoms and prolactin level elevation noted at baseline. It achieved these results with minimal (0%-27%) D2 occupancy 12 hours after the last dose. Study of the additional subjects revealed that quetiapine does give rise to transiently high (58%-64%) D2 occupancy 2 to 3 hours after a single dose that then decreases to minimal levels in 12 hours. CONCLUSIONS: Quetiapine shows a transiently high D2 occupancy, which decreases to very low levels by the end of the dosing interval. Quetiapine's low D2 occupancy can explain its freedom from extrapyramidal symptoms and prolactin level elevation. The data suggest that transient D2 occupancy may be sufficient for its antipsychotic effect. Future studies controlling for nonpharmacological effects as well as activities on other receptors will be necessary to confirm this suggestion. PMID- 10839334 TI - Children's well-being 11 years after the Chornobyl catastrophe. AB - BACKGROUND: The psychological effects of technological disasters have rarely been studied in children. This study assessed the aftermath of the 1986 Chornobyl disaster in children evacuated to Kyiv from the contaminated zone surrounding the nuclear power facility. METHODS: In 1997, we evaluated three hundred 10- to 12 year-old children in Kyiv who were in utero or infants at the time of the disaster and who had resided near Chornobyl (evacuees) and 300 sex-matched homeroom classmates who had never lived in a radiation-contaminated area. Response rates were 92% (evacuees) and 85% (classmates). Data were obtained from children, mothers, and teachers using standard measures of well-being and risk factors for childhood psychopathology. The children also received physical examinations and basic blood tests. RESULTS: The evacuees and classmates perceived their mental health similarly except for Chornobyl-related anxiety symptoms and perceived scholastic competence. No differences were found on the Iowa Conners' Teacher Rating Scale. Although the physical examination and blood test results were normal, the evacuee mothers rated their children's well-being as significantly worse, especially with respect to somatic symptoms on the Children's Somatization Inventory and Child Behavior Checklist. The most important risk factors for these ratings were maternal somatization and Chornobyl related stress. CONCLUSIONS: Given the multiple stressful experiences to which evacuee families were exposed, the small differences in the children's self reports suggest that there are protective factors in the lives of these children. The trauma experienced by the mothers was reflected in their perceptions of their children's well-being, particularly somatic symptoms, but was not transmitted to the children themselves. PMID- 10839335 TI - Melatonin suppression by light in euthymic bipolar and unipolar patients. AB - BACKGROUND: Previous studies have suggested that bipolar patients are supersensitive to light suppression of melatonin and that this may be a trait marker for genetic vulnerability. The present study was an attempt to replicate and extend this observation. Propranolol hydrochloride effects were compared with light effects because of the documented influence of beta-adrenergic receptors on melatonin production. Nighttime levels of corticotropin and cortisol were also examined as potential trait vulnerability markers. METHODS: Melatonin levels in euthymic bipolar patients (n= 29) were tested before and after 500-lux light was administered between 2 and 4 AM and on a separate night in the dark. Results were compared with those of a group of patients with unipolar depression (n= 24) and with those of a group of non-psychiatrically ill control subjects (n= 50). Lithium effects and propranolol effects were tested in subgroups. RESULTS: No group differences were seen in light suppression among bipolar patients, unipolar patients, and controls; an analysis of the whole group did not reveal differences in propranolol effect, differences in corticotropin or cortisol levels, or evidence for a lithium effect. However, patients with bipolar I affective disorder showed the following: (1) significantly lower melatonin levels on the light night, at baseline and following light exposure; and (2) a later peak time for melatonin on the dark night. CONCLUSIONS: The general hypothesis of increased light sensitivity in bipolar patients was not supported. However, melatonin secretion abnormalities were confirmed in the subgroup with bipolar I disorder. Further assessments of circadian rhythm disruption as a vulnerability marker in bipolar illness are indicated. PMID- 10839336 TI - The effects of electroconvulsive therapy on memory of autobiographical and public events. AB - BACKGROUND: Retrograde amnesia is the most persistent cognitive adverse effect of electroconvulsive therapy (ECT); however, it is not known whether ECT has differential effects on autobiographical vs impersonal memories. This study examined the short- and long-term effects of differing forms of ECT on memory of personal and impersonal (public) events. METHODS: Fifty-five patients with major depression were randomly assigned to right unilateral (RUL) or bilateral (BL) ECT, each at either low or high electrical dosage. The Personal and Impersonal Memory Test was administered by blinded raters at baseline, during the week after ECT, and at the 2-month follow-up. Normal controls were tested at matched intervals. RESULTS: Shortly after ECT, patients recalled fewer events and event details than controls, with the deficits most marked for impersonal compared with personal events. Bilateral ECT caused more marked amnesia for events and details than RUL ECT, and especially for impersonal memories. These effects were independent of electrical dosage and clinical outcome. At the 2-month follow-up, patients had reduced retrograde amnesia, but continued to show deficits in recalling the occurrence of impersonal events and the details of recent impersonal events. CONCLUSIONS: The amnestic effects of ECT are greatest and most persistent for knowledge about the world (impersonal memory,) compared with knowledge about the self (personal memory), for recent compared with distinctly remote events, and for less salient events. Bilateral ECT produces more profound amnestic effects than RUL ECT, particularly for memory of impersonal events. PMID- 10839337 TI - Retrograde amnesia with electroconvulsive therapy: characteristics and implications. PMID- 10839338 TI - Congruence of diagnoses 2 years after a first-admission diagnosis of psychosis. AB - BACKGROUND: Diagnostic changes may reflect evolution of an illness, emergence of newly disclosed information, or unreliability of assessment. This study evaluates the stability of research diagnoses in a heterogeneous first-admission sample with psychosis. METHODS: A group of 547 subjects initially diagnosed with a psychosis were reassessed 6 and 24 months after enrollment. The DSM-IV consensus diagnoses were formulated by psychiatrists blind to previous research diagnoses. The analysis focuses on agreement over time and the effects of demographic, family history, and clinical variables on the shift from a nonschizophrenia diagnosis to schizophrenia. RESULTS: Seventy-two percent of 6- and 24-month diagnoses were congruent. The most temporally consistent 6-month categories were schizophrenia (92%), bipolar disorder (83%), and major depression (74%); the least stable were psychosis not otherwise specified (44%), schizoaffective disorder (36%), and brief psychosis (27%). The most frequent shift in diagnosis at 24 months was to schizophrenia spectrum (n=45). These 45 subjects had a similar illness course after 6 months as the 171 subjects in this category at both assessments, but their prior clinical functioning was better. Risk factors predicting change to a schizophrenia spectrum diagnosis include facility variables (schizophrenia diagnosis, longer stays, and given antipsychotic medication on hospital discharge); prehospital features (psychotic > or =3 months before admission, poorer adolescent adjustment, lifetime substance disorder); and negative symptoms. CONCLUSIONS: Changes in diagnosis, particularly to schizophrenia, are mostly attributable to the evolution of the illness. Rigid adherence to DSM-IV requirements may have led to underdiagnosis of schizophrenia. The findings support the need for a longitudinally based diagnostic process in incidence samples. PMID- 10839339 TI - Prevalence of depression and its treatment in an elderly population: the Cache County study. AB - BACKGROUND: Previous estimates of the prevalence of geriatric depression have varied. There are few large population-based studies; most of these focused on individuals younger than 80 years. No US studies have been published since the advent of the newer antidepressant agents. METHODS: In 1995 through 1996, as part of a large population study, we examined the current and lifetime prevalence of depressive disorders in 4,559 nondemented individuals aged 65 to 100 years. This sample represented 90% of the elderly population of Cache County, Utah. Using a modified version of the Diagnostic Interview Schedule, we ascertained past and present DSM-IV major depression, dysthymia, and subclinical depressive disorders. Medication use was determined through a structured interview and a "medicine chest inventory." RESULTS: Point prevalence of major depression was estimated at 4.4% in women and 2.7% in men (P= .003). Other depressive syndromes were surprisingly uncommon (combined point prevalence, 1.6%). Among subjects with current major depression, 35.7% were taking an antidepressant (mostly selective serotonin reuptake inhibitors) and 27.4% a sedative/hypnotic. The current prevalence of major depression did not change appreciably with age. Estimated lifetime prevalence of major depression was 20.4% in women and 9.6% in men (P<.001), decreasing with age. CONCLUSIONS: These estimates for prevalence of major depression are higher than those reported previously in North American studies. Treatment with antidepressants was more common than reported previously, but was still lacking in most individuals with major depression. The prevalence of subsyndromal depressive symptoms was low, possibly because of unusual characteristics of the population. PMID- 10839340 TI - Prepulse inhibition of the startle response in men with schizophrenia: effects of age of onset of illness, symptoms, and medication. AB - BACKGROUND: Prepulse inhibition of the startle reflex response refers to the ability of a weak prestimulus to transiently inhibit the response to a closely following strong sensory stimulus. This effect represents an operational index of sensorimotor gating and is found to be deficient in schizophrenia. Prepulse inhibition deficits in schizophrenia seem to be partially normalized by typical antipsychotics and more fully by some atypical antipsychotics. Early onset of schizophrenia, particularly in men, has been associated with abnormal brain maturation, profound neuropsychological deficits, and less responsiveness to antipsychotic medication. We evaluated the effects of the age of onset of illness, current positive and negative symptoms, and the type of medication (typical vs atypical) on prepulse inhibition of the startle response in schizophrenia. METHODS: Thirty-eight male schizophrenic patients and 20 healthy male controls underwent testing for prepulse inhibition of the acoustic startle response. RESULTS: Earlier onset of illness was associated with reduced prepulse inhibition, while adult onset of illness was not. No significant relationships occurred between current symptoms and prepulse inhibition. Patients given typical, but not atypical, antipsychotics exhibited less prepulse inhibition compared with healthy controls. CONCLUSION: Early onset of illness is associated with profound deficits in prepulse inhibition of the startle response in men with schizophrenia. PMID- 10839341 TI - Recurrent depression and stressful life events. PMID- 10839342 TI - Hippocampal and superior temporal gyrus volume in first-episode schizophrenia. PMID- 10839343 TI - Waltzing mice add a new twist to the function of EphB2. PMID- 10839344 TI - Does motor protein intelligence contribute to neuronal polarity? PMID- 10839345 TI - Orphanin FQ/Nociceptin and the mystery of pain. PMID- 10839346 TI - Neurotrophic rescue of photoreceptors: are Muller cells the mediators of survival? PMID- 10839347 TI - Highwire, rpm-1, and futsch: balancing synaptic growth and stability. PMID- 10839348 TI - Putting the spinal cord together again. PMID- 10839349 TI - What rest in flies can tell us about sleep in mammals. PMID- 10839350 TI - Experience-dependent development of spinal motor neurons. AB - Locomotor activity in many species undergoes pronounced alterations in early postnatal life, and environmental cues may be responsible for modifying this process. To determine how these events are reflected in the nervous system, we studied rats reared under two different conditions-the presence or absence of gravity-in which the performance of motor operations differed. We found a significant effect of rearing environment on the size and complexity of dendritic architecture of spinal motor neurons, particularly those that are likely to participate in postural control. These results provide evidence that neurons subserving motor function undergo activity-dependent maturation in early postnatal life in a manner analogous to sensory systems. PMID- 10839351 TI - Autoimmunity to gephyrin in Stiff-Man syndrome. AB - Stiff-Man syndrome (SMS) is a rare disease of the central nervous system (CNS) characterized by chronic rigidity, spasms, and autoimmunity directed against synaptic antigens, most often the GABA-synthesizing enzyme glutamic acid decarboxylase (GAD). In a subset of cases, SMS has an autoimmune paraneoplastic origin. We report here the identification of high-titer autoantibodies directed against gephyrin in a patient with clinical features of SMS and mediastinal cancer. Gephyrin is a cytosolic protein selectively concentrated at the postsynaptic membrane of inhibitory synapses, where it is associated with GABA(A) and glycine receptors. Our findings provide new evidence for a close link between autoimmunity directed against components of inhibitory synapses and neurological conditions characterized by chronic rigidity and spasms. PMID- 10839352 TI - Highwire regulates synaptic growth in Drosophila. AB - The formation, stabilization, and growth of synaptic connections are dynamic and highly regulated processes. The glutamatergic neuromuscular junction (NMJ) in Drosophila grows new boutons and branches throughout larval development. A primary walking behavior screen followed by a secondary anatomical screen led to the identification of the highwire (hiw) gene. In hiw mutants, the specificity of motor axon pathfinding and synapse formation appears normal. However, NMJ synapses grow exuberantly and are greatly expanded in both the number of boutons and the extent and length of branches. These synapses appear normal ultrastructurally but have reduced quantal content physiologically. hiw encodes a large protein found at presynaptic terminals. Within presynaptic terminals, HIW is localized to the periactive zone surrounding active zones; Fasciclin II (Fas II), which also controls synaptic growth, is found at the same location. PMID- 10839353 TI - Regulation of presynaptic terminal organization by C. elegans RPM-1, a putative guanine nucleotide exchanger with a RING-H2 finger domain. AB - Presynaptic terminals contain highly organized subcellular structures to facilitate neurotransmitter release. In C. elegans, the typical presynaptic terminal has an electron-dense active zone surrounded by synaptic vesicles. Loss of-function mutations in the rpm-1 gene result in abnormally structured presynaptic terminals in GABAergic neuromuscular junctions (NMJs), most often manifested as a single presynaptic terminal containing multiple active zones. The RPM-1 protein has an RCC1-like guanine nucleotide exchange factor (GEF) domain and a RING-H2 finger. RPM-1 is most similar to the Drosophila presynaptic protein Highwire (HIW) and the mammalian Myc binding protein Pam. RPM-1 is localized to the presynaptic region independent of synaptic vesicles and functions cell autonomously. The temperature-sensitive period of rpm-1 coincides with the time of synaptogenesis. rpm-1 may regulate the spatial arrangement, or restrict the formation, of presynaptic structures. PMID- 10839354 TI - rpm-1, a conserved neuronal gene that regulates targeting and synaptogenesis in C. elegans. AB - Little is known of mechanisms regulating presynaptic differentiation. We identified rpm-1 in a screen for mutants with defects in patterning of a presynaptic marker at certain interneuronal synapses. The predicted RPM-1 protein contains zinc binding, RCC1, and other conserved motifs. In rpm-1 mutants, mechanosensory neurons fail to accumulate tagged vesicles, retract synaptic branches, and ectopically extend axons. Some motor neurons branch and overgrow; others show altered synaptic organization. Expression of RPM-1 in the presynaptic mechanosensory neurons is sufficient to rescue phenotypes in these cells. Certain rpm-1 phenotypes are temperature sensitive, revealing that RPM-1 function can be bypassed by maintaining mutants at the permissive temperature at stages commensurate with synapse formation in wild-type animals. These results indicate that RPM-1 functions cell autonomously during synaptogenesis to regulate neuronal morphology. PMID- 10839355 TI - Drosophila Futsch/22C10 is a MAP1B-like protein required for dendritic and axonal development. AB - Here we report the description of the Drosophila gene futsch, which encodes a protein recognized by the monoclonal antibody 22C10 that has been widely used to visualize neuronal morphology and axonal projections. The Futsch protein is 5327 amino acids in length. It localizes to the microtubule compartment of the cell and associates with microtubules in vitro. The N- and C-terminal domains of Futsch are homologous to the vertebrate MAP1B microtubule-associated protein. The central domain of the Futsch protein is highly repetitive and shows sequence similarity to neurofilament proteins of which no Drosophila homologs have been reported. Loss-of-function analyses demonstrate that during embryogenesis Futsch is necessary for dendritic and axonal growth. Gain-of-function analyses demonstrate a functional interaction of Futsch with other MAPs. In addition, we show that during development, futsch expression is negatively regulated in nonneuronal tissues. PMID- 10839356 TI - Drosophila Futsch regulates synaptic microtubule organization and is necessary for synaptic growth. AB - We present evidence that Futsch, a novel protein with MAP1B homology, controls synaptic growth at the Drosophila neuromuscularjunction through the regulation of the synaptic microtubule cytoskeleton. Futsch colocalizes with microtubules and identifies cytoskeletal loops that traverse the lateral margin of select synaptic boutons. An apparent rearrangement of microtubule loop architecture occurs during bouton division, and a genetic analysis indicates that Futsch is necessary for this process. futsch mutations disrupt synaptic microtubule organization, reduce bouton number, and increase bouton size. These deficits can be partially rescued by neuronal overexpression of a futsch MAP1B homology domain. Finally, genetic manipulations that increase nerve-terminal branching correlate with increased synaptic microtubule loop formation, and both processes require normal Futsch function. These data suggest a common microtubule-based growth mechanism at the synapse and growth cone. PMID- 10839358 TI - Radial glial identity is promoted by Notch1 signaling in the murine forebrain. AB - In vertebrates, Notch signaling is generally thought to inhibit neural differentiation. However, whether Notch can also promote specific early cell fates in this context is unknown. We introduced activated Notch1 (NIC) into the mouse forebrain, before the onset of neurogenesis, using a retroviral vector and ultrasound imaging. During embryogenesis, NIC-infected cells became radial glia, the first specialized cell type evident in the forebrain. Thus, rather than simply inhibiting differentiation, Notch1 signaling promoted the acquisition of an early cellular phenotype. Postnatally, many NIC-infected cells became periventricular astrocytes, cells previously shown to be neural stem cells in the adult. These results suggest that Notch1 promotes radial glial identity during embryogenesis, and that radial glia may be lineally related to stem cells in the adult nervous system. PMID- 10839357 TI - rax, Hes1, and notch1 promote the formation of Muller glia by postnatal retinal progenitor cells. AB - We are interested in the mechanisms of glial cell development in the vertebrate central nervous system. We have identified genes that can direct the formation of glia in the retina. rax, a homeobox gene, Hes1, a basic helix-loop-helix gene, and notch1, a transmembrane receptor gene, are expressed in retinal progenitor cells, downregulated in differentiated neurons, and expressed in Muller glia. Retroviral transduction of any of these genes resulted in expression of glial markers. In contrast, misexpression of a dominant-negative Hes1 gene reduced the number of glia. Cotransfection of rax with reporter constructs containing the Hes1 or notch1 regulatory regions led to the upregulation of reporter transcription. These data suggest a regulatory heirarchy that controls the formation of glia at the expense of neurons. PMID- 10839359 TI - A developmentally regulated switch directs regenerative growth of Schwann cells through cyclin D1. AB - Sciatic nerve axons in cyclin D1 knockout mice develop normally, become properly ensheathed by Schwann cells, and appear to function normally. However, in the Wallerian degeneration model of nerve injury, the mitotic response of Schwann cells is completely inhibited. The mitotic block is Schwann cell autonomous and developmentally regulated. Rescue analysis (by "knockin" of cyclin E) indicates that D1 protein, rather than regulatory elements of the D1 gene, provides the essential Schwann cell function. Genetic inhibition of the Schwann cell cycle shows that neuronal responses to nerve injury are surprisingly independent of Schwann cell mitotic responses. Even axonal regrowth into the distal zone of a nerve crush injury is not markedly impaired in cyclin D1-/- mice. PMID- 10839360 TI - EphB2 guides axons at the midline and is necessary for normal vestibular function. AB - Mice lacking the EphB2 receptor tyrosine kinase display a cell-autonomous, strain specific circling behavior that is associated with vestibular phenotypes. In mutant embryos, the contralateral inner ear efferent growth cones exhibit inappropriate pathway selection at the midline, while in mutant adults, the endolymph-filled lumen of the semicircular canals is severely reduced. EphB2 is expressed in the endolymph-producing dark cells in the inner ear epithelium, and these cells show ultrastructural defects in the mutants. A molecular link to fluid regulation is provided by demonstrating that PDZ domain-containing proteins that bind the C termini of EphB2 and B-ephrins can also recognize the cytoplasmic tails of anion exchangers and aquaporins. This suggests EphB2 may regulate ionic homeostasis and endolymph fluid production through macromolecular associations with membrane channels that transport chloride, bicarbonate, and water. PMID- 10839361 TI - A critical role for a Rho-associated kinase, p160ROCK, in determining axon outgrowth in mammalian CNS neurons. AB - We tested the contribution of the small GTPase Rho and its downstream target p160ROCK during the early stages of axon formation in cultured cerebellar granule neurons. p160ROCK inhibition, presumably by reducing the stability of the cortical actin network, triggered immediate outgrowth of membrane ruffles and filopodia, followed by the generation of initial growth cone-ike membrane domains from which axonal processes arose. Furthermore, a potentiation in both the size and the motility of growth cones was evident, though the overall axon elongation rate remained stable. Conversely, overexpression of dominant active forms of Rho or ROCK was suggested to prevent initiation of axon outgrowth. Taken together, our data indicate a novel role for the Rho/ROCK pathway as a gate critical for the initiation of axon outgrowth and the control of growth cone dynamics. PMID- 10839362 TI - Regulation of neuregulin signaling by PSD-95 interacting with ErbB4 at CNS synapses. AB - Neuregulins (NRGs) and their receptors, the ErbB protein tyrosine kinases, are essential for neuronal development, but their functions in the adult CNS are unknown. We report that ErbB4 is enriched in the postsynaptic density (PSD) and associates with PSD-95. Heterologous expression of PSD-95 enhanced NRG activation of ErbB4 and MAP kinase. Conversely, inhibiting expression of PSD-95 in neurons attenuated NRG-mediated activation of MAP kinase. PSD-95 formed a ternary complex with two molecules of ErbB4, suggesting that PSD-95 facilitates ErbB4 dimerization. Finally, NRG suppressed induction of long-term potentiation in the hippocampal CA1 region without affecting basal synaptic transmission. Thus, NRG signaling may be synaptic and regulated by PSD-95. A role of NRG signaling in the adult CNS may be modulation of synaptic plasticity. PMID- 10839363 TI - SNAREs contribute to the specificity of membrane fusion. AB - Intracellular membrane fusion is mediated by the formation of a four-helix bundle comprised of SNARE proteins. Every cell expresses a large number of SNARE proteins that are localized to particular membrane compartments, suggesting that the fidelity of vesicle trafficking might in part be determined by specific SNARE pairing. However, the promiscuity of SNARE pairing in vitro suggests that the information for membrane compartment organization is not encoded in the inherent ability of SNAREs to form complexes. Here, we show that exocytosis of norepinephrine from PC12 cells is only inhibited or rescued by specific SNAREs. The data suggest that SNARE pairing does underlie vesicle trafficking fidelity, and that specific SNARE interactions with other proteins may facilitate the correct pairing. PMID- 10839364 TI - The role of selective transport in neuronal protein sorting. AB - To assess whether selective microtubule-based vesicle transport underlies the polarized distribution of neuronal proteins, we expressed green fluorescent protein- (GFP-) tagged chimeras of representative axonal and dendritic membrane proteins in cultured hippocampal neurons and visualized the transport of carrier vesicles containing these proteins in living cells. Vesicles containing a dendritic protein, transferrin receptor (TfR), were preferentially transported into dendrites and excluded from axons. In contrast, vesicles containing the axonal protein NgCAM (neuron-glia cell adhesion molecule) were transported into both dendrites and axons. These data demonstrate that neurons utilize two distinct mechanisms for the targeting of polarized membrane proteins, one (for dendritic proteins) based on selective transport, the other (for axonal proteins) based on a selectivity "filter" that occurs downstream of transport. PMID- 10839365 TI - Long-term depression of the cerebellar climbing fiber--Purkinje neuron synapse. AB - In classic Marr-Albus-Ito models of cerebellar function, coactivation of the climbing fiber (CF) synapse, which provides massive, invariant excitation of Purkinje neurons (coding the unconditioned stimulus), together with a graded parallel fiber synaptic array (coding the conditioned stimulus) leads to long term depression (LTD) of parallel fiber-Purkinje neuron synapses, underlying production of a conditioned response. Here, we show that the supposedly invariant CF synapse can also express LTD. Brief 5 Hz stimulation of the CF resulted in a sustained depression of CF EPSCs that did not spread to neighboring parallel fiber synapses. Like parallel fiber LTD, CF LTD required postsynaptic Ca2+ elevation, activation of group 1 mGluRs, and activation of PKC. CF LTD is potentially relevant for models of cerebellar motor control and learning and the developmental conversion from multiple to single CF innervation of Purkinje neurons. PMID- 10839366 TI - Ca2+-induced Ca2+ release supports the relay mode of activity in thalamocortical cells. AB - Ca2+ ions play an important role during rhythmic bursting of thalamocortical neurons within sleep. The function of Ca2+ during the tonic relay mode of these neurons during wakefulness is less clear. Here, we report that tonic activity in thalamocortical cells results in an increase in the intracellular Ca2+ concentration and subsequent release of Ca2+ from intracellular stores mediated via ryanodine receptors (RyRs). Blockade of Ca2+ release shifted the regular firing of single action potentials toward the generation of spike clusters. Regular spike firing and intracellular Ca2+ release thus appear to be functionally coupled in a positive feedback manner, thereby supporting the relay mode of thalamocortical cells during wakefulness. Regulatory influences may be coupled to this system via the cyclic ADP ribose pathway. PMID- 10839367 TI - Drosophila CRY is a deep brain circadian photoreceptor. AB - cry (cryptochrome) is an important clock gene, and recent data indicate that it encodes a critical circadian photoreceptor in Drosophila. A mutant allele, cry(b), inhibits circadian photoresponses. Restricting CRY expression to specific fly tissues shows that CRY expression is needed in a cell-autonomous fashion for oscillators present in different locations. CRY overexpression in brain pacemaker cells increases behavioral photosensitivity, and this restricted CRY expression also rescues all circadian defects of cry(b) behavior. As wild-type pacemaker neurons express CRY, the results indicate that they make a striking contribution to all aspects of behavioral circadian rhythms and are directly light responsive. These brain neurons therefore contain an identified deep brain photoreceptor, as well as the other circadian elements: a central pace-maker and a behavioral output system. PMID- 10839368 TI - A TIMELESS-independent function for PERIOD proteins in the Drosophila clock. AB - The mutation timeless(UL) generates 33 hr rhythms, prolonged nuclear localization of PERIOD/TIMELESS(UL) protein complexes, and protracted derepression of period (per) and timeless (tim) transcription. Light-induced elimination of TIM(UL) from nuclear PER/TIM(UL) complexes gives strong downregulation of per and tim expression. Thus, in the absence of TIM, nuclear PER can function as a potent negative transcriptional regulator. Two additional studies support this role for PER: (1) Drosophila expressing PER that constitutively localizes to nuclei produce dominant behavioral arrhythmicity, and (2) constitutively nuclear PER represses dCLOCK/CYCLE-mediated transcription of per in cultured cells without TIM. Conversion of PER/TIM heterodimers to nuclear PER proteins appears to be required to complete transcriptional repression and terminate each circadian molecular cycle. PMID- 10839369 TI - A cellular mechanism for the bidirectional pain-modulating actions of orphanin FQ/nociceptin. AB - Orphanin FQ/nociceptin (OFQ/N) and its receptor share substantial structural features and cellular actions with classic opioid peptides and receptors, but have distinct pharmacological profiles and behavioral effects. Currently there is an active debate about whether OFQ/N produces hyperalgesia or analgesia. Using a well-defined brainstem pain-modulating circuit, we show that OFQ/N can cause either an apparent hyperalgesia by antagonizing mu opioid-induced analgesia or a net analgesic effect by reducing the hyperalgesia during opioid abstinence. It presumably produces these two opposite actions by inhibiting two distinct groups of neurons whose activation mediates the two effects of opioid administration. OFQ/N antagonism of the hyperalgesia may have significance for the treatment of opioid withdrawal and sensitized pain. PMID- 10839370 TI - Peripheral demyelination and neuropathic pain behavior in periaxin-deficient mice. AB - The Prx gene in Schwann cells encodes L- and S-periaxin, two abundant PDZ domain proteins thought to have a role in the stabilization of myelin in the peripheral nervous system (PNS). Mice lacking a functional Prx gene assemble compact PNS myelin. However, the sheath is unstable, leading to demyelination and reflex behaviors that are associated with the painful conditions caused by peripheral nerve damage. Older Prx-/- animals display extensive peripheral demyelination and a severe clinical phenotype with mechanical allodynia and thermal hyperalgesia, which can be reversed by intrathecal administration of a selective NMDA receptor antagonist We conclude that the periaxins play an essential role in stabilizing the Schwann cell-axon unit and that the periaxin-deficient mouse will be an important model for studying neuropathic pain in late onset demyelinating disease. PMID- 10839371 TI - Modification of glial-neuronal cell interactions prevents photoreceptor apoptosis during light-induced retinal degeneration. AB - Prolonged or high-intensity exposure to visible light leads to photoreceptor cell death. In this study, we demonstrate a novel pathway of light-induced photoreceptor apoptosis involving the low-affinity neurotrophin receptor p75 (p75NTR). Retinal degeneration upregulated both p75NTR and the high-affinity neurotrophin receptor TrkC in different parts of Muller glial cells. Exogenous neurotrophin-3 (NT-3) increased, but nerve growth factor (NGF) decreased basic fibroblast growth factor (bFGF) production in Muller cells, which can directly rescue photoreceptor apoptosis. Blockade of p75NTR prevented bFGF reduction and resulted in both structural and functional photoreceptor survival in vivo. Furthermore, the absence of p75NTR significantly prevented light-induced photoreceptor apoptosis. These observations implicate glial cells in the determination of neural cell survival, and suggest functional glial-neuronal cell interactions as new therapeutic targets for neurodegeneration. PMID- 10839372 TI - Intubation and survival in severe paediatric blunt head injury. AB - The majority of severe childhood injuries are due to head injuries. We studied the impact of emergency intubation in a cohort of children suffering severe blunt head trauma. A 10-year retrospective case note analysis was performed on 176 children (age < 16 years) with severe blunt head trauma (abbreviated injury scale > or =4) in Southern Finland, who required intensive care in a level 1 trauma centre, or who died despite initiation of life supporting measures at the scene. Children in whom emergency intubation was performed either at the scene, or in the emergency room (ER) were analysed. Of the 59 children who fulfilled the study criteria, 20 had an isolated head injury. Most injuries (56/59) were caused by road traffic accidents. Field-intubation was performed in 24 children, and emergency intubation in the ERs of regional hospitals or the level 1 trauma centre, in 13 and 22 children respectively. Mortality was 54.2% (32/59), and was highest in children intubated in regional hospital ERs or in the field. Children intubated at the scene or in the ER of regional hospitals, had significantly worse AIS (head/neck), injury severity score (ISS), and Glasgow coma (GCS) scores than those children intubated in the ER of the level 1 trauma centre. Survival was better in field-intubated children compared with those intubated in regional hospital ERs, despite similar trauma scores (p = 0.05). It is concluded that although children with severe (AIS > or =4) head injury who require emergency intubation have a high overall mortality, field-intubation may improve survival, compared with 'scoop and run' with BLS airway management and deferred emergency intubation. PMID- 10839373 TI - Aetiologic diagnosis of ischaemic stroke in the emergency department: relevance for triage and clinical management. AB - Effective strategies for the aetiologic diagnosis in patients with ischaemic stroke can be implemented based on simple clinical criteria and instrumental tests which can be performed in a modern emergency room (ER) within 24 hours from admission. This may bear prognostic and therapeutic relevance for patients with acute stroke. Therefore, in this study we set out to establish the feasibility and accuracy of the aetiologic diagnosis of ischaemic stroke in an ER. A total of 136 consecutive patients (mean age 72+/-10 years, 60 females) with first ever ischaemic stroke admitted during 1996-1997 were evaluated with assessment of clinical features, CT scan, ECG, ultrasonography of the extracranial arteries, transthoracic echocardiography, and, in selected patients, transoesophageal echocardiography. Patients were classified into two major categories defined as stroke of determined origin and stroke of undetermined origin (a stroke with two or more possible causes or with a negative evaluation), according to the TOAST criteria. Ninety-six patients were considered affected by stroke of determined origin (70.5%), (22.7% with large artery atherosclerosis, 19.1% with cardioembolic stroke, 26.4% with lacunar stroke and 1.4% with other aetiology). The remaining 40 patients (29.4%) had stroke of undetermined origin: of these, 13 patients (9.5%) had a totally negative evaluation, 15 patients (12.5%) showed cardioembolism among the two or more possible causes of stroke and seven patients (5.1%) had atherothrombotic or lacunar aetiology. Additional work-up with transoesophageal echocardiography succeeded in demonstrating aortic embolism in five patients (3.6%; i.e. four patients with aortic plaques more than 4 mm in thickness and one patient with ulcerated plaques). In conclusion, the subtype classification system for ischaemic stroke allowed the aetiological diagnosis in 70.5% of patients while in the ER. Stroke of undetermined origin represented one third of patients in a consecutive population with acute onset neurologic deficit of ischaemic origin. In approximately half of the patients with negative standard evaluation, cardiogenic or aortic arch embolic sources could be identified by transoesophageal echocardiography. Thus, the latter is indicated in patients with stroke of undetermined origin with negative first-line evaluation in order to identify embolic sources in the aortic arch. PMID- 10839374 TI - Has an intensified treatment in the ambulance of patients with acute severe left heart failure improved the outcome? AB - The aim of this study was to evaluate short- and long-term outcome prior to and after the introduction of a more intensified treatment in the ambulance of patients with acute severe heart failure. Consecutive patients with acute severe heart failure transported by the mobile coronary care unit (MCCU) in the community of Goteborg prior to and after the introduction of an intensified treatment (nitroglycerine, continuous positive airway pressure (CPAP) and furosemide). One hundred and fifty-eight patients were evaluated during each period. The median age was 77 and 76.5 years, respectively, and 52% and 42% were women. The proportion of patients given nitroglycerine in the ambulance was 4% and 68% in the two periods; the proportion of patients treated with furosemide was 13% and 84%, respectively. CPAP was used in less than 1% during period 1 and in 91% during period 2. On admission of the ambulance 60% had fulminant pulmonary oedema during period 1 versus 78% during period 2 (p<0.0001). On admission to hospital the opposite was found, 93% during period 1 versus 76% during period 2 (p<0.0001). The median serum creatinine kinase (CK-MB) maximum activity was 13 microkat/l during period 1 and 8 microkat/l during period 2 (p = 0.007). However, the mortality during the first year remained high during both periods (39.2% and 35.8%, p = 0.64). It is concluded that a more intensive treatment in the ambulance of patients with acute severe heart failure seems to have resulted in an improvement in symptoms during transport and less myocardial damage. However, no significant improvement in long-term mortality was observed. PMID- 10839375 TI - Effectiveness of imaging studies in acute appendicitis: a simplified decision model. AB - Computed tomography (CT) and ultrasonography (US) have been proposed as non invasive diagnostic aids in patients with suspected acute appendicitis. Currently, clear guidelines about the indications to use these techniques are lacking. Using the concepts of decision analysis, a model was created to calculate the effectiveness of US and CT in patients with clinically suspected appendicitis. The perspective chosen was the health of individual patients. The model makes use of published data and provides a critical threshold probability (ctp). Decisions in individual cases can be made by comparing ctp with the clinical probability of disease. The calculated ctp-values for US and CT were 0.58 and 0.74, respectively. In other words, on average, US (CT) is indicated only if the expert clinician considers the probability of disease to be smaller than 58% (74%). It is concluded that in patients with suspected acute appendicitis, selective rather than routine use of imaging studies is recommended. The exact value of the decision threshold should be determined in function of the local situation. PMID- 10839376 TI - Upper limb in injuries in dry ski slope skiing--a continuing problem. AB - The aim of this study was to assess how common injuries to the upper limbs are from dry ski slope skiing, to document the pattern of injury to the upper limbs from dry ski slope skiing and to discuss preventative measures. Skier's experience was also determined. The study was carried out on 174 patients attending the accident and emergency department over a period of 3 years who had sustained injuries from dry ski slope skiing. Prospective registration was used to collect the data. Sixty-seven per cent of the patients had received instructions prior to skiing. Upper limb injuries outnumbered lower limb injuries by a ratio of 7.9 to 1. Thumb injuries outnumbered other injuries in the upper limb. The mechanism of injury was due to a fall in 94.2% of patients. It is concluded that the lattice network on dry ski slopes significantly contributes to the platform of injuries to the upper limbs, and ways in which the lattice network may be improved are recommended. PMID- 10839377 TI - The 'three in one' block as locoregional analgesia in an emergency department. AB - The 'three in one' block is a well-known locoregional technique for per- and post operative anaesthesia. This 'three in one' block was investigated in an emergency department with three combinations of local anaesthetics as a locoregional way of pain relief for patients with fractures of the upper femur. A double-blind prospective study was conducted with 61 consecutive patients. A good regression of the linear visual analogue scale (LVAS), good haemodynamic stability and great patient satisfaction were found for all groups of patients at different times after installation of the block. Side effects were few and are avoidable by reducing the amount of local anaesthetic given to elderly patients with a low bodyweight. The technique failed in only four patients due to technical reasons. It was concluded that the 'three in one' block is a fast, safe, reliable and easy technique for pain relief in an emergency department. Good analgesia was achieved for patients with fractures of the upper femur with no need to change the position of the patient. PMID- 10839378 TI - A national survey of the Polish population's cardiopulmonary resuscitation knowledge. AB - The teaching of cardiopulmonary resuscitation (CPR) has had a positive public health impact beyond resuscitating cardiac arrest victims to include recognition of signs and symptoms and prevention of heart attacks. To study the general knowledge and degree of preparedness in the Republic of Poland, a national survey was conducted. The survey was designed to assess general knowledge and concern regarding CPR training in the Polish population. Complete demographic information was collected to determine any difference in responses in the population. The results indicate that (1) approximately 75% of the Polish population has received CPR training; (2) the majority of the surveyed population assess their CPR ability as inadequate; (3) the surveyed population believe that CPR training needs to be expanded and improved. Taking into consideration results of the CPR knowledge assessment questions it can be concluded that CPR knowledge in Poland is low and a standardized curriculum for CPR training is needed. PMID- 10839379 TI - Victims of violence in an emergency department. AB - The aim of this study was to identify the risk profile of the victims of violent accidents. During 1997, we studied 1100 patients suffering from injuries produced by violent actions and treated in the emergency department of the largest teaching hospital in the Canary Islands, Spain. Seventy per cent of the patients were male and 30% were female. The mean age was 28 and 30 years, respectively. In the distribution by age, the main group was that between 18 and 36 years. Seventy five per cent of violent accidents were public, 20% domestic and 5% remain unknown. The face-neck was the anatomical location with the highest number of severe injuries, followed by the upper extremities (42% vs. 26%, p<0.001). Eight per cent of the injured men presented severe injuries, in contrast with 1% of the women (p<0.001). Most patients (94%) were discharged from hospital after being treated. None of the victims died as a result of violence. The admission of victims occurred predominantly during the night hours and 35% of the patients were treated during the weekend. It is concluded that the typical victim of violent accidents in an emergency department of a Spanish teaching hospital is a young male with severe injuries, which occurred during late hours at weekends. PMID- 10839380 TI - Does witnessed cardiopulmonary resuscitation alter perceived stress in accident and emergency staff? AB - The aim of this study was to ascertain if the presence of patients' relatives during cardiopulmonary resuscitation altered perceived symptoms of stress in accident and emergency personnel participating in resuscitation attempts. An anonymous structured questionnaire survey of all accident and emergency staff participating in non-traumatic adult cardiopulmonary resuscitation was designed to elicit symptoms of an acute stress reaction within 24 hours based on ICD-10 diagnostic criteria. One hundred and fourteen staff replies were received, a reply rate of 89%. Twenty-five replies had two or more symptoms of an acute stress reaction. The grade or role of the staff member had no influence on the presence of stress symptoms. There was no difference in rates of reporting between staff resuscitating in the presence or absence of relatives. It is concluded that the presence of relatives witnessing resuscitation attempts does not affect self-reported stress symptoms in staff participating in resuscitation attempts. PMID- 10839381 TI - Helicopter emergency medical service in Canary Islands, Spain. AB - This is a report on our first 2 years' experience of operating a helicopter emergency medical service in the Canary Islands, Spain. The two advanced life support helicopters are staffed full time by a physician and a nurse. For the transport protocol, inter-hospital transport patients (secondary missions) were classified into three groups: group A, minor illnesses or injuries; group B, modified or middle critical condition; and group C, critical condition. On-scene patients (primary missions) were also divided into critical and non-critical condition. Cardiovascular and respiratory stabilization were necessary before transport. One thousand and fifty-four patients were transported, 19% of whom were primary missions and 81% of whom were secondary missions. Thirty per cent of the first group were in critical condition. The distribution of secondary missions was group A 16%, group B 44% and group C 40%. In group C, 60% of patients were mechanically ventilated, 70% needed cardiovascular drug support and 84% needed stabilization before transport. Thirty-two per cent were trauma patients and 12% neonates. The overall mortality rate was 0.8%. The cost per mission was US$2300. In the interests of safety and rationalization of the use of resources, transport of non-critical patients should be reduced. The presence of a trained physician and nursing crew and stabilization before transport could be responsible for the low mortality rate. PMID- 10839382 TI - Certification of an emergency department according to UNI EN ISO 9002 criteria. AB - Over the past decade, the philosophy of total quality has appeared in all fields of industry in Japan and the USA. This philosophy is now present all over Europe as well, Italy included. In 1993 the Arcispedale Santa Maria Nuova of Reggio Emilia, a town in Northern Italy, began to apply the total quality management philosophy, thus embarking on a course of continuous quality improvement. The present article describes the main steps which led to an external accredited team granting the emergency department of this hospital a quality system certificate in 1997 according to the international quality system requirements UNI EN ISO 9002. The importance of this work lies in the fact than an industrial quality standardization system, whose requirements have little in common with a traditional view of medical practice, can nevertheless be applied to a public health care department. PMID- 10839383 TI - Advice on writing an abstract for a scientific meeting and on the evaluation of abstracts by selection committees. PMID- 10839384 TI - Surgical emphysema of the face and neck due to repeated Valsalva following a laceration to the buccal mucosa. PMID- 10839385 TI - Spontaneous hepatic rupture in pregnancy. AB - The HELLP-syndrome (haemolysis, elevated liver enzymes, low platelets) is associated with pre-eclampsia and may cause subcapsular liver haematomas. When hepatic rupture occurs the mortality of mother and unborn is high. Rupture remains a surgical emergency with control of bleeding based on trauma principles. We report a case and discuss the diagnosis and management. PMID- 10839386 TI - Foreign body aspiration in childhood: management algorithm. PMID- 10839387 TI - More data about patients who leave emergency department without being seen by a doctor in European hospitals. PMID- 10839388 TI - Microsurgical strategies in 74 patients for restoration of dynamic depressor muscle mechanism: a neglected target in facial reanimation. PMID- 10839389 TI - A magnetic resonance imaging investigation of potential subclinical complications after in situ cranial bone graft harvest. AB - The in situ harvest of cranial bone grafts, when assessed by clinical observation, has been shown to be a safe procedure when performed by experienced surgeons. However, might there be adverse sequelae from bone graft harvest that are not severe enough to produce observable changes and thus are missed by studies that rely on clinical outcomes? This study was designed to determine the incidence of "clinically silent" complications following routine bone graft harvest, such as punctate intracranial bleeding and cerebral contusions. This prospective clinical study included 20 patients. All patients had in situ cranial bone graft harvest performed by a single surgeon and underwent postharvest magnetic resonance imaging within 24 hours. Coronal computed tomographic scans were found to be the most helpful in the preoperative determination of donor sites. The average area of bone graft harvest was just under 14 cm2 (range, 3 to 30 cm2). No patients who had not previously undergone skull surgery were noted to have full-thickness breaches. Three patients identified with full-thickness breaches at surgery had harvest sites located in areas in which a previous craniotomy had occurred, suggesting that extreme care be used when harvesting bone from this subset of patients. All 20 postoperative magnetic resonance scans were reviewed by a neuroradiologist and determined to be completely normal. In conclusion, not only is in situ cranial bone graft harvest a safe procedure as assessed by clinical outcomes, but no subclinical complications were identified by post-operative magnetic resonance imaging. PMID- 10839390 TI - An evaluation of hard palate mucosa graft as a lining material in alar reconstruction: a 7-year experience applied to the full-thickness alar defect. AB - The authors present their experience with 25 hard palate mucosa grafts used as lining material in the reconstruction of full-thickness alar defects. Good "take" was obtained in 22 grafts; the other three grafts incurred necrosis of the overriding skin flaps and postoperative infection. Degree of shrinkage was 11 to 15 percent of grafted size in patients with the type of defect that did not include the alar margin; shrinkage was 26 to 35 percent in patients with the type that included more than 50 percent of the alar margin. In all patients who had a good graft take, the nasal cavities were maintained and there was no nasal obstruction or collapsing during strong breathing. The healing time of the palate donor site varied from 7 days to 5 weeks, depending on the size of the defect. No patients experienced any symptoms at the donor site after healing. The authors concluded that hard palate mucosa can be considered a useful material in alar reconstruction because of the ease in graft harvesting and its support features. When the defect is large enough to involve the total unilateral ala nasi, even though the degree of postoperative shrinkage is comparatively high, hard palate mucosa may be the most suitable material to ensure good take of the graft and less possibility of donor-site morbidity. PMID- 10839391 TI - Treatment of facial wounds with botulinum toxin A improves cosmetic outcome in primates. AB - Surgeons have constantly sought to achieve the most aesthetic scar. A major factor determining the final cosmetic appearance of a cutaneous scar is the tension acting on the wound edges during the healing phase. Since Theodor Kocher pioneered the alignment of skin incisions with Langer's lines in 1892, surgical techniques that attempt to overcome closing tension have become standard. Yet, no treatment has been available to minimize underlying muscle contractions, which are the major cause of this tension. Botulinum toxin A is a potent drug that produces temporary muscular paralysis when injected locally. It has proven to be safe and effective in the treatment of a variety of disorders, including hyperkinetic facial lines. The objective of this randomized, double-blind, placebo-controlled primate study was to investigate the efficacy of a single injection of botulinum toxin A to improve the cosmetic appearance of cutaneous scars. Symmetric pairs of standardized excisions were performed on either side of the forehead of six primates. The half foreheads were randomized to the botulinum toxin A treatment side versus the placebo injection side. A panel of three blinded facial surgeons assessed the cosmetic appearance of the mature scars 3 months postoperatively. The wounds that had been immobilized with botulinum toxin A were rated as significantly better in appearance than the control wounds (p < 0.01). Histologic examination confirmed that all scars were mature. Blinded, randomized, placebo-controlled human clinical trials are presently under way at the Mayo Clinic. PMID- 10839392 TI - Sentinel lymph node biopsy for melanoma: experience with 234 consecutive procedures. AB - Sentinel lymph node biopsy is increasingly used to identify occult metastases in regional lymph nodes of patients with melanoma. Selection of patients for sentinel lymph node biopsy and subsequent lymphadenectomy is an area of debate. The purpose of this study was to describe a large clinical series of these biopsies for cutaneous melanoma and to identify patients most likely to gain useful clinical information from sentinel lymph node biopsy. The Indiana University Melanoma Program computerized database was queried to identify all patients who underwent this procedure for clinically localized cutaneous melanoma. It was performed using preoperative technetium Tc 99m lymphoscintigraphy and isosulfan blue dye. Pertinent demographic, surgical, and histopathologic data were recorded. Univariate and multivariate logistic regression and classification table analyses were performed to identify clinical variables associated with sentinel node and nonsentinel node positivity. In total, 234 biopsy procedures were performed to stage 291 nonpalpable regional lymph node basins. Mean Breslow's thickness was 2.30 mm (2.08 mm for negative sentinel lymph node biopsy, 3.18 mm for positive). The mean number of sentinel nodes removed was 2.17 nodes per basin (range, 1 to 8). Forty-seven of 234 melanomas (20.1 percent) and 50 of 291 basins (17.2 percent) had a positive biopsy. Positivity correlated with AJCC tumor stage: T1, 3.6 percent; T2, 8.1 percent; T3, 27.4 percent; T4, 44 percent. By univariate logistic regression, Breslow's thickness (p = 0.003, continuous variable), ulceration (p = 0.003), mitotic index > or = 6 mitoses per high power field (p = 0.008), and Clark's level (p = 0.04) were significantly associated with sentinel lymph node biopsy result. By multivariate analysis, only Breslow's thickness (p = 0.02), tumor ulceration (p = 0.02), and mitotic index (p = 0.02) were significant predictors of biopsy positivity. Classification table analysis showed the Breslow cutpoint of 1.2 mm to be the most efficient cutpoint for sentinel lymph node biopsy result (p = 0.0004). Completion lymphadenectomy was performed in 46 sentinel node positive patients; 12 (26.1 percent) had at least one additional positive nonsentinel node. Nonsentinel node positivity was marginally associated with the presence of multiple positive sentinel nodes (p = 0.07). At mean follow-up of 13.8 months, four of 241 sentinel node-negative basins demonstrated same-basin recurrence (1.7 percent). Sentinel lymph node biopsy is highly reliable in experienced hands but is a low-yield procedure in most thin melanomas. Patients with melanomas thicker than 1.2 mm or with ulcerated or high mitotic index lesions are most likely to have occult lymph node metastases by sentinel lymph node biopsy. Completion therapeutic lymphadenectomy is recommended after positive biopsy because it is difficult to predict the presence of positive nonsentinel nodes. PMID- 10839393 TI - Outcome following implantation of a peripheral nerve stimulator in patients with chronic nerve pain. AB - This study evaluated the usefulness of the implanted peripheral nerve stimulator in patients with pain following injury to a peripheral nerve. The patient sample (n = 17) consisted of 7 men and 10 women with a mean age of 48 years (SD = 18 years). The mean follow-up time since implantation of the stimulator was 21 months (SD = 15 months). Workers' compensation and/or litigation were involved in 11 cases. Peripheral nerve stimulators were placed in the upper extremity in 12 patients and in the lower extremity in 5 patients. Pain relief following implantation was rated as excellent by five patients, good by six patients, fair by four patients, and poor by two patients. A statistically significant decrease in reported pain level was found postoperatively (p < 0.0003). There was no statistically significant difference in postoperative pain level between men and women (p = 0.30), between cases involving workers' compensation or litigation and those not involving these issues (p = 1.0), or between patients who received an upper-extremity implant and those who received a lower-extremity implant (p = 0.56). Of the 12 patients who were unable to work before the operation, 6 returned to work after the operation. In conclusion, peripheral nerve stimulators can be useful in decreasing pain in carefully selected patients with severe neurogenic pain. PMID- 10839394 TI - Treatment of burn scar depigmentation by carbon dioxide laser-assisted dermabrasion and thin skin grafting. AB - Permanent depigmentation occasionally develops after deep partial-thickness and full-thickness burn injuries, which heal by secondary intention. This problem can be solved by dermabrasion and thin split-thickness skin grafting. However, mechanical dermabrasion is a bloody procedure that risks exposing medical professionals to infectious diseases transmitted by blood products, and it is difficult to assess the extent of tissue ablation. In this study, dermabrasion of depigmented burn scar area was performed by using flash-scanned carbon dioxide laser treatment, followed by thin split-thickness skin grafting. This method was applied to 13 patients on whom burn scar depigmentation sites were located as follows: two in the facial area, four on the trunk, and seven on the extremities. Skin graft take was excellent in all patients except for one. The follow-up period for these patients ranged from 1 to 12 months, with an average of 8 months. Repigmentation appeared soon after grafting, and no depigmentation occurred again in the treated areas. In conclusion, depigmented burn scar areas can be dermabraded in a short time; depth of tissue ablation can be well controlled; and a bloodless and smooth raw surface can be created by using a flash-scanned carbon dioxide laser. These raw surfaces sustain thin skin grafts well. PMID- 10839395 TI - Silicone gel-filled breast implant integrity: a retrospective review of 478 consecutively explanted implants. AB - Concern has been expressed over the long-term integrity of silicone gel breast implants. There are no large series representing experience with these implants outside of the United States. A retrospective case note review of explanted silicone breast implants was performed; 478 implants have been explanted during the past 11 years and relate to the use of these devices since 1971. Loss of implant integrity was not simply related to its age in vivo. Failure was more likely with implants of the late 1970s and early 1980s (second generation) and with subpectoral placement. Implant failure was independent of capsular contracture as the indication for removal (p = 0.09). There is no evidence that the currently used textured silicone gel breast implants are subject to the same loss of integrity as previous examples of these devices. The life span of these implants, the first of which are approaching 10 years in vivo, is at present unknown. Information concerning the integrity of silicone gel breast implants is essential in the current climate for counseling of both new and old implant recipients. PMID- 10839396 TI - Neophalloplasty in female-to-male transsexuals with the island tensor fasciae latae flap. AB - In the past 60 years, several different procedures have attempted to achieve a postoperative neophallus that is as aesthetic and as functional as possible after penile amputation or sex reassignment. Recently, with improvements in free tissue transfer and microvascular technique, many free flap procedures have been developed with the goal of an aesthetically acceptable neophallus of adequate bulk that enables urination in a standing position and sexual intercourse, with minimal functional and aesthetic donor-site defects. Most authors currently agree that the method of choice for penile reconstruction is microsurgical free tissue transfer, although it does not always fulfill all of the aforementioned goals in a predictable manner. In fact, complete urethroplasty, penile rigidity, and donor site disfigurement remain challenges, thus making this operation one of the most difficult in plastic surgery. The vascular anatomy of the lateral circumflex femoral artery, which we studied in 1991 with the anatomic dissection of 27 cadavers, gave us the idea to use a long tensor fasciae latae neurovascular island flap as a donor source for neophalloplasty. Grounds for the procedure and its surgical planning have been carefully evaluated with 10 additional fresh cadaver dissections. Since 1991, we have performed five neophalloplasties using this procedure; all patients were female-to-male transsexuals. In four cases, the healing was uneventful; in one case, there was a marginal necrosis of the flap because of poor venous drainage, probably from a twisting of the pedicle. The island tensor fasciae latae provides a safe and sensate flap for phalloplastic procedure and leaves a less conspicuous donor scar. PMID- 10839397 TI - Surgical treatment of the cleft foot. AB - Between 1970 and 1997, we treated a total of 32 cleft feet in 21 patients (11 male and 10 female). We classified cases of cleft foot on the basis of the number of central ray deficiencies. Fourteen patients with 22 cleft feet were followed up for more than 1 year postoperatively (9 feet had no or one central ray deficiency, and 13 feet had two or three central ray deficiencies). The mean follow-up period was 8.8 years. The objective of this study was to evaluate the results of operative treatment of cleft foot. We evaluated the results of three methods: simple closure of the cleft, application of a double-pedicled flap, and insertion of a silicone block. Cosmetic complications, including widening of the foot, hypertrophic scarring, pigmentation of the grafted skin, and overlapping of the toes, were observed in patients with two or three central ray deficiencies. Few functional complications were observed: None of the patients experienced gait disturbances, although one patient complained of pain following walking. Roentgenography showed that the distance between the first and fifth metatarsals was 86 percent of that of the contralateral foot. When treating patients with no or one central ray deficiency, satisfactory results can be expected with simple closure of the cleft. However, in patients with two or three central ray deficiencies, it is difficult to obtain satisfactory results with simple closure of the cleft or application of a double-pedicled flap. Therefore, silicone block insertion to correct the defect is recommended when there is more than one central ray deficiency. PMID- 10839398 TI - Skeletal muscle reinnervation by reduced axonal numbers results in whole muscle force deficits. AB - Patients sustaining a peripheral nerve injury will frequently experience residual muscle weakness after muscle reinnervation, even if the nerve repair is performed under optimal circumstances to allow rapid muscle reinnervation. The mechanisms responsible for this contractile dysfunction remain unclear. It is hypothesized that after peripheral nerve injury and repair, a reduced number of axons are available for skeletal muscle reinnervation that results in whole muscle force and specific force deficits. A rat model of peroneal nerve injury and repair was designed so that the number of axons available for reinnervation could be systematically reduced. In adult rats, the peroneal nerve to the extensor digitorum longus muscle was either left intact (sham group, n = 8) or divided and repaired with either 50 percent (R50 group, n = 7) or 100 percent (R100 group, n = 8) of the axons in the proximal stump included in the repair. Four months after surgery, maximal tetanic isometric force was measured and specific force was calculated for each animal. Mean tetanic isometric force for extensor digitorum longus muscles from R50 rats (2765.7 +/- 767.6 mN) was significantly lower than sham (4082.8 +/- 196.5 mN) and R100 (3729.0 +/-370.2 mN) rats (p < 0.003). Mean specific force calculations revealed significant deficits in both the R100 (242.1 +/- 30 kN/m2) and R50 (190.6 +/- 51.8 kN/m2) rats compared with the sham animals (295.9 +/- 14 kN/m2) (p < 0.0005). These data support our hypothesis that after peripheral nerve injury and repair, reinnervation of skeletal muscle by a reduced number of axons results in a reduction in tetanic isometric force and specific force. The greater relative reduction in specific force compared with absolute force production after partial nerve repair may indicate that a population of residual denervated muscle fibers is responsible for this deficit. PMID- 10839399 TI - Sutural expansion osteogenesis for management of the bony-tissue defect in cleft palate repair: experimental studies in dogs. AB - A series of experimental studies on sutural expansion osteogenesis for management of the bony-tissue defect in cleft palate repair was performed between 1995 and 1997. Forty-five young dogs in weaning were used in four experiments that were divided into two parts. Part I probed the possibility of closing the surgically constructed hard palate cleft not only with mucoperiosteum but also with bony tissue by the technique of sutural expansion of lateral palatine sutures. Part II explored the possibility of pushing the palatine bone posteriorly and advancing the maxillary segment anteriorly by transverse palatine suture expansion. In Part I, a ring-shaped suture expander made of nickel-titanium shape memory alloy was used to expand the lateral suture of palatine bones. Expansion forces of 200 G, 360 G, and 480 G were used for the first experiment. A force of 360 G was chosen for two other experiments; this force is equivalent to the distraction rate of 0.5 mm per day of a jackscrew device. The ring-shaped suture expander was opened and its two feet were fixed in the medial sides of residual horizontal plates of the palatine bones immediately after a hard palate cleft was constructed surgically under endotracheal general anesthesia. At the eighth postoperative day, under the traction of 360 G, the two sides of the 8-mm-wide hard palate cleft were brought into contact with each other, and 8 or 9 days later the closed palatal cleft had healed completely with mucosal tissue. This experiment was repeated twice and yielded the same results. Sutural expansion osteogenesis was evaluated physically, fluorescently, histologically, and ultrastructurally to examine the deposition of the regenerated bone in the suture areas. Additionally, the influence of sutural expansion osteogenesis of the palatal bones on other facial bones was also studied cephalometrically. In Part II, a bow-shaped suture expander made of nickel-titanium shape memory alloy was applied to expand either the left or the right side of the transverse palatal suture of each of the experimental dogs. At the postoperative week 4 to 6, the maxillary segment was moved forward 5 to 6 mm on the expanded side, and the palatal bone was pushed backward 5 mm. The changes of bone position were assessed radiographically and cephalometrically. Tissue response of circum-maxillary sutures was examined histologically. These experiments led to the following conclusions: (1) Bony closure of the surgically constructed hard palate cleft with a ring-shaped suture expander made of nickel-titanium shape memory alloy is possible. (2) Anterior advancement of the maxillary segment and posterior lengthening of the hard palate using a bow-shaped suture expander made of nickel-titanium shape memory alloy applied at the palatomaxillary suture (transverse palatal suture) of the hard palate are also possible. Thus, in humans, a new approach for cleft palate repair may be a worthwhile investigation. PMID- 10839400 TI - Gene expression of TGF-beta, TGF-beta receptor, and extracellular matrix proteins during membranous bone healing in rats. AB - Poorly healing mandibular fractures and osteotomies can be troublesome complications of craniomaxillofacial trauma and reconstructive surgery. Gene therapy may offer ways of enhancing bone formation by altering the expression of desired growth factors and extracellular matrix molecules. The elucidation of suitable candidate genes for therapeutic intervention necessitates investigation of the endogenously expressed patterns of growth factors during normal (i.e., successful) fracture repair. Transforming growth factor beta1 (TGF-beta1), its receptor (Tbeta-RII), and the extracellular matrix proteins osteocalcin and type I collagen are thought to be important in long-bone (endochondral) formation, fracture healing, and osteoblast proliferation. However, the spatial and temporal expression patterns of these molecules during membranous bone repair remain unknown. In this study, 24 adult rats underwent mandibular osteotomy with rigid external fixation. In addition, four identically treated rats that underwent sham operation (i.e., no osteotomy) were used as controls. Four experimental animals were then killed at each time point (3, 5, 7, 9, 23, and 37 days after the procedure) to examine gene expression of TGF-beta1 and Tbeta-RII, osteocalcin, and type I collagen. Northern blot analysis was used to compare gene expression of these molecules in experimental animals with that in control animals (i.e., nonosteotomized; n = 4). In addition, TGF-beta1 and T-RII proteins were immunolocalized in an additional group of nine animals killed on postoperative days 3, 7, and 37. The results of Northern blot analysis demonstrated a moderate increase (1.7 times) in TGF-beta1 expression 7 days postoperatively; TGF-beta1 expression returned thereafter to near baseline levels. Tbeta-RII mRNA expression was downregulated shortly after osteotomy but then increased, reaching a peak of 1.8 times the baseline level on postoperative day 9. Osteocalcin mRNA expression was dramatically downregulated shortly after osteotomy and remained low during the early phases of fracture repair. Osteocalcin expression trended slowly upward as healing continued, reaching peak expression by day 37 (1.7 times the control level). In contrast, collagen type IalphaI mRNA expression was acutely downregulated shortly after osteotomy, peaked on postoperative days 5, and then decreased at later time points. Histologic samples from animals killed 3 days after osteotomy demonstrated TGF-beta1 protein localized to inflammatory cells and extracellular matrix within the fracture gap, periosteum, and peripheral soft tissues. On postoperative day 7, TGF-beta1 staining was predominantly localized to the osteotomized bone edges, periosteum, surrounding soft tissues, and residual inflammatory cells. By postoperative day 37, complete bony healing was observed, and TGF-beta1 staining was localized to the newly formed bone matrix and areas of remodeling. On postoperative day 3, Tbeta-RII immunostaining localized to inflammatory cells within the fracture gap, periosteal cells, and surrounding soft tissues. By day 7, Tbeta-RII staining localized to osteoblasts of the fracture gap but was most intense within osteoblasts and mesenchymal cells of the osteotomized bone edges. On postoperative day 37, Tbeta-RII protein was seen in osteocytes, osteoblasts, and the newly formed periosteum in the remodeling bone. These observations agree with those of previous in vivo studies of endochondral bone formation, growth, and healing. In addition, these results implicate TGF-beta1 biological activity in the regulation of osteoblast migration, differentiation, and proliferation during mandibular fracture repair. Furthermore, comparison of these data with gene expression during mandibular distraction osteogenesis may provide useful insights into the treatment of poorly healing fractures because distraction osteogenesis has been shown to be effective in the management of these difficult clinical cases. PMID- 10839401 TI - The effect of superpulsed carbon dioxide laser energy on keloid and normal dermal fibroblast secretion of growth factors: a serum-free study. AB - An in vitro model was used to determine the effect of superpulsed CO2 laser energy on normal dermal and keloid-producing fibroblast proliferation and release of growth factors. Growth factors assayed included basic fibroblast growth factor (bFGF) and transforming growth factor beta1 (TGF-beta1). bFGF is mitogenic, inhibits collagen production, and stabilizes cellular phenotype. TGF-beta1 stimulates growth and collagen secretion and is thought to be integral to keloid formation. Growth in a serum-free medium allowed measurement of these growth factors without confounding variables. Keloid and normal dermal fibroblasts cell lines were established from facial skin samples using standard explant techniques. Samples consisted of three separate keloid and three separate normal dermal fibroblast cell lines. Cells were used at passage 4 to seed 24-well trays at a concentration of 6 x 10(4) cells per milliliter in serum-free medium. At 48 hours, 18.8 percent of each cell well was exposed to a fluence of 2.4, 4.7, and 7.3 J/cm2 using the superpulsed CO2 laser. Cell viability and counts were established at four time points: 0 (time of superpulsed CO2 laser treatment), 24, 72, and 120 hours. Supernatants were collected and assessed for bFGF and TGF beta1 using a sandwich enzyme immunoassay. All cell lines demonstrated logarithmic growth through 120 hours (conclusion of experiment), with a statistically significant shorter population doubling time for keloid fibroblasts (p < 0.05). Use of the superpulsed CO2 laser shortened population doubling times relative to that of controls; the differences were statistically significant in keloid dermal fibroblasts when fluences of 2.4 and 4.7 J/cm2 were used (p < 0.05 and 0.01, respectively). bFGF was present in greater levels in normal dermal fibroblasts than in keloid dermal fibroblasts. Application of superpulsed CO2 demonstrated a trend toward increased bFGF secretion in both fibroblast types; the increase was significant in the keloid group at 4.7J/cm2. A consistent trend in suppression of TGF-beta1 was seen in both groups exposed to superpulsed CO2, with the maximal effect occurring at 4.7 J/cm2. Serum-free culture sustains logarithmic cell growth and allows growth factor measurement without confounding variables from serum-containing media. Superpulsed CO2 enhances fibroblast replication and seems to stimulate bFGF secretion and to inhibit TGF-beta1 secretion. Given the function of these growth factors, the application of superpulsed CO2 may support normalized wound healing. These findings may explain the beneficial effects of laser resurfacing on a cellular level and support the use of superpulsed CO2 in the management of keloid scar tissue. PMID- 10839402 TI - Soft-tissue augmentation with injectable alginate and syngeneic fibroblasts. AB - Tissue engineering, a field that combines polymer scaffolds with isolated cell populations to create new tissue, may be applied to soft-tissue augmentation-an area in which polymers and cell populations have been injected independently. We have developed an inbred rat model in which the subcutaneous injection of a hydrogel, a form of polymer, under vacuum permits direct comparison of different materials in terms of both histologic behavior and their ability to maintain the specific shape and volume of a construct. Using this model, we compared three forms of calcium alginate, a synthetic hydrogel, over an 8-week period-standard alginate that was gelled following injection into animals (alginate post-gel), standard alginate that was gelled before injection into animals (alginate pre gel) and alginate-RGD, to which the cell adhesion tripeptide RGD was linked covalently (RGD post-gel). Parallel groups that included cultured syngeneic fibroblasts suspended within each of these three gels were also evaluated (alginate post-gel plus cells, alginate pre-gel plus cells, and RGD post-gel plus cells). The study used 54 inbred Lewis rats (n = 9 for each of the six groups). Construct geometry was optimally maintained in the alginate post-gel group in which 58 percent of the original volume was preserved at 8 weeks and increased to 88 percent at 8 weeks when syngeneic fibroblasts were included within the gel. Volume was not as well preserved in the RGD post-gel group (25 percent of original volume at 8 weeks), but again increased when syngeneic fibroblasts were included (41 percent of original volume at 8 weeks). Maintenance of volume was poorest in the alginate pre-gel group (31 percent of original volume at 8 weeks) and failed to be augmented by the addition of fibroblasts (19 percent of original volume at 8 weeks). Histologically, the gel remained a uniform sheet surrounded by a fibrous capsule in the alginate post-gel groups. In the alginate pre-gel and RGD post-gel groups, there was significant ingrowth of a fibrovascular stroma into the gel with fragmentation of the construct. In constructs in which syngeneic fibroblasts were included, cells were visualized throughout the gel but did not extend processes or appear to contribute to new tissue formation. Material compression testing indicated that the alginate and RGD post-gel constructs became stiffer over a 12-week period, particularly in the cell containing groups. Our results suggest that calcium alginate could be a suitable agent for soft-tissue augmentation when gelled subcutaneously following injection. The addition of syngeneic fibroblasts enhanced the ability of the gel to maintain the volume of a construct; this seems to be mediated by increased gel stiffness rather than by de novo tissue formation. Our animal model, in combination with material testing data, permits rigorous comparison of different materials used for soft-tissue augmentation. PMID- 10839403 TI - Efficacy of intervention strategies in a brachial plexus global avulsion model in the rat. AB - The treatment of brachial plexus avulsion lesions invariably involves the use of neurotization procedures. Although some of these therapeutic strategies have been used for the past 20 years to restore selective function to the injured extremity, the individual efficacy of these nerve transfers has not been measured objectively, thereby rendering the prognostication of outcomes for these techniques a major problem. Using a true global avulsion model, the present study compares outcomes of the various neurotization procedures for the first time. The strength of this experimental model is that each nerve transfer tested leads to a common terminal pathway involving a single target-namely, the biceps muscle. Thus, quantitative measurements of biceps restoration will provide strong clues to the power of axonal regrowth of that particular motor pool. This study also introduces the Terzis grooming test, a modified behavioral test that can be quantified and that can provide an overall functional scale in the assessment of outcome. Thirty-five Sprague-Dawley rats were divided into seven groups, with each group testing a different motor donor for biceps reinnervation. The ipsilateral brachial plexus was globally avulsed in all animals, with the exception of the ipsilateral C7 group, and the respective motor donor coapted in an end-to-end fashion to the musculocutaneous nerve. Functional outcomes were measured by the Terzis grooming test, electromyography, biceps muscle force measurements, motor end plate counts, and quantitative axonal morphometry. The values of the different parameters were expressed as a standard score on a common scale. The relative standings of each group on each parameter were compared. Superior outcome was observed in the phrenic, the hypoglossal, and the ipsilateral C7 groups. PMID- 10839404 TI - Use of a modified oxygen microelectrode and laser-Doppler flowmetry to monitor changes in oxygen tension and microcirculation in a flap. AB - Flap failure is a clinical problem in free tissue transfer, and there is no reliable device for monitoring the tissue. Differentiating between an arterial occlusion and venous congestion is also a problem. A study was undertaken to monitor viability in a pedicled groin flap and to compare two different monitoring methods. The oxygen tension in the flap, measured with a modified Clark-type microelectrode (tip diameter = 3 to 8 microm; 90 percent response within 2.6 +/- 0.5 seconds), was compared with changes in blood flow in the flap, measured with a laser-Doppler probe. In 11 Sprague-Dawley rats, the changes in oxygen tension and blood flow in the pedicled groin flap were studied after clamping and subsequent reperfusion of the artery or vein. After occlusion of the artery to the flap, oxygen tension decreased to a stable value (i.e., the recording level remained unchanged for 30 seconds), from 19.7 +/- 1.8 to 0.3 +/- 0.1 mmHg, after 193 +/-25 seconds; blood flow decreased to a stable value, from 117 +/- 21 to 54 +/- 18 perfusion units, after 26 +/- 6 seconds. Clamping of the vein resulted in a decrease in oxygen tension, from 17.1 +/- 1.8 to 1.4 +/- 0.7 mmHg, after 416 +/- 67 seconds, and blood flow decreased to a stable value, from 90 +/- 14 to 35 +/- 6 perfusion units, after 107 +/- 27 seconds. The results of this study show that there is a difference in oxygen tension and blood flow responses between arterial and venous occlusion and that it may be possible with both methods to distinguish arterial from venous occlusion. However, although oxygen tension measurements are slightly slower in response than laser-Doppler measurements, the values are more reliable as a diagnostic tool for interpretation of a vessel occlusion. PMID- 10839405 TI - Ectopic bone formation after temporal muscle transposition for facial paralysis. AB - This report describes the case of a patient with congenital bilateral facial paralysis in whom ectopic bone formation developed following temporal muscle transposition. Ectopic bone formation was first noticed 4 years after surgery. Whether the ossification is a result of the transfer of periosteum or the osteogenic capacity of muscular tissue is still unknown. PMID- 10839406 TI - Shoulder reconstruction by latissimus dorsi myocutaneous flap based on the serratus branch after advanced soft-tissue sarcoma excision. PMID- 10839407 TI - Mirror foot. AB - Mirror image polydactyly is a rare congenital abnormality that may occur in isolation, or in association with multiple congenital anomalies. A case of unilateral mirror foot with an ipsilateral short, broad tibia is described. The clinical, radiologic, and operative findings are presented, and current theories of embryo-pathogenesis are reviewed. PMID- 10839408 TI - Intraneural perineurioma involving the median nerve. AB - Intraneural perineurioma is a rare clinical entity, which tends to affect major nerve trunks in the upper extremities. On light microscopy, numerous pseudo-onion bulb structures having a central clear area are surrounded by concentric layers of eosinophilic elongate cells having spindled nuclei. Immunohistochemistry of concentric cells stains positive for epithelial membrane antigen but negative for S100 protein. Because of the small number of cases, no consensus has been made on proper treatment of this entity. Although none of the patients who have had excision of tumor with nerve grafting have had sensory nerve recovery, we believe each patient should be individualized until more data are available regarding this tumor. PMID- 10839409 TI - Excision of the submandibular gland by an intraoral approach. AB - To improve the outcome in patients with benign diseases of the submandibular gland, we have developed an entirely intraoral technique for excision of the submandibular gland. This procedure is anatomically safe and can be performed with minimal morbidity. We believe the essential surgical steps are as follows: (1) infiltration with Xylocaine plus epinephrine with an adequate waiting period for hemostasis; (2) careful identification of the submandibular duct/lingual nerve relationship; (3) anterior retraction of the mylohyoid muscle to expose the superficial lobe; (4) superiorly directed, extraoral, manipulation of the submandibular gland; and (5) close and blunt dissection to the gland laterally to avoid injury to the facial artery and vein. PMID- 10839410 TI - Method for intraoperative positioning of the nipple-areola complex in vertical scar reduction mammaplasty. AB - The desired shape and position of the nipple-areola complex may be difficult to achieve in vertical scar reduction mammaplasty when using the standard technique of preoperative marking of the so-called mosque-shaped areolar pattern of excision. We describe our modified approach of intraoperative final positioning of the nipple-areola complex by hiding the nipple-areola complex behind the closed vertical incision. Individual positioning at the final part of the operation allows for more predictable results and also for a calculated lower positioning, which enables balance of the potential bottoming-out of the breast, particularly in previously large ptotic breasts. We believe that this modification helps to further improve the results of vertical scar reduction mammaplasty by adding more possibilities for shaping and "last-minute" modifications intraoperatively. PMID- 10839411 TI - Prepuce pollicization: a reminder of an alternate donor. PMID- 10839412 TI - Protective clear shield for dorsal surgical wound sites in the rat. PMID- 10839413 TI - Cutaneous melanoma of the head and neck: advances in evaluation and treatment. PMID- 10839414 TI - Cleft palate repair at 3 to 7 months of age. AB - We report the speech outcome in 90 children with complete unilateral cleft lip and palate who underwent soft palate repair either between 3 and 7 months of age (n = 40) or later than 7 months of age (n = 50). In all patients, palatoplasty was performed by one of two experienced surgeons using a modification of the Furlow technique, and speech evaluations were conducted using the Pittsburgh Weighted Values for Speech Symptoms Associated with Velopharyngeal Incompetence by two speech pathologists with high inter-rater reliability. There were no differences between the groups with respect to resonance, nasal air emission, and articulation. Velopharyngeal function, as measured by the total speech score, was similar between the two groups of patients, as were the rates of secondary pharyngoplasty. These results suggest that very early closure of the soft palate may not offer significant benefit over repair later in infancy with respect to speech outcome. PMID- 10839415 TI - Microsurgical replantation of the amputated nose. AB - A case of successful replantation of the nose is presented. Two arteries and one vein were anastomosed, providing a stable framework for direct revascularization of the amputated nasal segment. This resulted in complete survival of the nose, with an excellent aesthetic result. However, despite successful microsurgical arterial and venous repair, significant postoperative blood loss still occurred as a result of anticoagulation. In cases of the amputation of specialized structures, the improved functional and cosmetic result obtained with replantation must be weighed against the risk of blood-borne disease transmission when postoperative transfusion is required. Recognizing the potential need for postoperative transfusion in these cases is important in allowing the surgeon to exercise appropriate judgment in deciding whether replantation should be performed. PMID- 10839416 TI - The anti-aging revolution: an evolving role for plastic surgery. PMID- 10839417 TI - Saline-filled breast implant safety and efficacy: a multicenter retrospective review. AB - Our center undertook an industry-funded, outcomes-based, multicenter, retrospective review to evaluate the safety and efficacy of saline-filled breast implants. Our review was part of a pre-market approval review process mandated by the U.S. Food and Drug Administration. The design of our review was modeled on a Plastic Surgery Educational Foundation outcomes study previously conducted by our center. For this study, several significant changes were made to our previous protocol, including improved patient tracking, stronger biostatistical support, and a mandatory 10-year minimum patient follow-up period. Physician-reported and patient-reported data on 450 patients with 882 saline-filled breast implants placed between January 1, 1980, and June 30, 1986, were obtained. Mean patient follow-up period was 13.0 years. Most implants (93.9 percent) were placed for breast augmentation. Seventy-four percent were placed in a submammary position; 25.6 percent, subpectorally; and 0.2 percent, subcutaneously. The overall complication rate was 20.2 percent. Reoperation for capsular contracture or implant deflation was necessary in 104 of 450 patients (23.1 percent). Deflation occurred in 73 implants (8.3 percent) and was underreported according to the physicians' record review alone. This deflation rate is higher than the 5.5 percent previously reported by our center. However, 26 of these 73 deflations (35.6 percent) occurred in a single cohort of patients at one center using Surgitek saline implants. If this cohort is excluded, the deflation rate drops to 5.8 percent, a figure more consistent with data published in the literature and found in our previous study. Of the 73 deflations, spontaneous deflation was reported for 50 (74.6 percent), and the remainder were iatrogenic. Actuarial survival of the non-Surgitek implants was 98.4 percent to 99.8 percent at 5 years and 96.9 percent to 98.9 percent at 10 years (95 percent confidence interval). Risk factors for implant deflation included the use of Surgitek saline-filled implants (odds ratio = 17.5, p < 0.01), use of Heyer-Schulte and Mentor model 1800 implants (odds ratio = 3.0, p < 0.01), and implant size greater than 450 cc (odds ratio = 1.01, p < 0.02). Risk factors for capsular contracture included submammary implant position (odds ratio = 2.05, p = 0.03) and implant size greater than 450 cc (odds ratio = 1.01, p < 0.01). Overall, satisfaction was high: 93 percent of patients were "satisfied" or "very satisfied" with their implants. As in our earlier study, risk factors for patient dissatisfaction were reconstruction after mastectomy (odds ratio = 7.6, p = 0.011), significant breast firmness (odds ratio = 6.2, p < 0.001), and patient desire for smaller implants (odds ratio = 3.0, p = 0.020). In conclusion, our review provides additional outcomes-based evidence that saline-filled breast implants remain a safe, effective alternative to silicone gel-filled models. PMID- 10839418 TI - Comparative lipoplasty analysis of in vivo-treated adipose tissue. AB - A comparative histologic and chemical analysis was undertaken of adipose tissue treated in vivo with traditional, ultrasound-assisted, and external ultrasound assisted lipoplasty. A series of six healthy women undergoing elective liposuction according to the superwet technique using a 1:1 infiltration ratio with the estimated quantity of fat to be removed was included in the study. Four separate regions on each patient were treated independently in vivo with traditional liposuction, internal ultrasound-assisted liposuction, or external ultrasound-assisted liposuction for 7 minutes. External massage was used as a control. Four separate specimens of adipose tissue from each patient were assessed for cellular disruption using blinded histologic evaluation. The remainder of tissue was centrifuged to separate the aqueous phase from the cellular components and then spectrophotometrically analyzed for creatinine kinase and glycerol 3-phosphate dehydrogenase activity as markers of cellular disruption. Histologic analysis confirmed 70 to 90 percent cellular disruption with internal ultrasound-assisted liposuction. Suction-assisted and external ultrasound-assisted liposuction showed 5 to 25 percent disruption, whereas massage controls showed only 5 percent. Only internal ultrasound-assisted liposuction showed 5 to 20 percent thermal liquefaction. Absorbance analysis showed creatine kinase activity (sigma units) greatest in ultrasound-exposed tissue. Both external and internal ultrasound-assisted liposuction gave creatine kinase levels 28 to 33 percent greater than suction-assisted liposuction, which varied only 10 percent from controls. Glycerol 3-phosphate dehydrogenase activity was 44 percent greater for internal ultrasound-assisted liposuction than that detected with suction-assisted liposuction. Glycerol 3-phosphate dehydrogenase activity with external ultrasound-assisted liposuction and massage did not vary much from each other, at only 14 percent and 11 percent activity compared with internal ultrasound-assisted liposuction, respectively. Histologic and enzyme analysis of the different types of liposuction and their effect on adipocyte cellular disruption revealed no significant effect of external ultrasound or massage on the adipocytes. Further experimental studies are necessary to evaluate the role and efficacy of alternative techniques for body contouring. PMID- 10839419 TI - Ultrasound-assisted lipectomy using the solid probe: a retrospective review of 100 consecutive cases. AB - Ultrasound-assisted lipectomy using the solid probe is a predictable and safe method that can yield excellent results. This method is particularly useful and is indicated when the subdermis must be approached to smooth out surface irregularities and/or to stimulate skin retraction. The authors present their method using the solid probe and a retrospective study of their first 100 consecutive cases. The indications for using the solid probe, its advantages and disadvantages, associated complications, and representative cases are presented. PMID- 10839420 TI - Occlusion following laser resurfacing promotes reepithelialization and wound healing. AB - One of the critical parameters that has not been examined carefully following laser skin resurfacing is the effect of eschar on the wound healing process. Because occlusive dressings minimize the occurrence of eschar, the present study was undertaken to evaluate the effect of occlusion following laser resurfacing. It is clear that CO2 lasers promote epidermal cell loss and variable amounts of dermal injury. To characterize the wound repair process after laser treatment, biopsy specimens were obtained 2 to 4 days after treatment. Specimens from 15 patients were examined; the preauricular biopsy specimens were paired such that one specimen was from skin that had been occluded and the other specimen (from the same patient) was from skin treated without occlusion. Skin specimens were examined by indirect immunofluorescence using antibodies to specific epidermal and dermal antigens. The results indicate that the keratinocytes that repopulate the epidermis migrate from the hair follicles and express keratin 17, an intermediate filament protein expressed in keratinocytes during the early stages of wound healing. The migration of keratin 17-expressing cells begins 48 hours following laser resurfacing in skin treated with occlusion, whereas cell migration from the follicles of skin treated without occlusion is delayed. In summary, occlusion promotes enhanced cell migration and diminished eschar formation, resulting in more rapid healing. PMID- 10839421 TI - Lengthening of the postoperative short nose: combined use of a gull-wing concha composite graft and a rib costochondral dorsal onlay graft. AB - Several causes of short nose are known: congenital anomaly, developmental problem, trauma, and various types of rhinoplasty-the postoperative short nose being one of the most difficult problems to correct in plastic surgery. Contracted skin envelope, tissue deficiency of cartilage and mucosal lining, and poor circulation make postoperative short nose difficult to lengthen and susceptible to recurrence. Thus, for effective lengthening and long-term maintenance of it, specific grafts should be used to supplement the missing lining and cartilage and a mechanical support also is needed to withstand the skin contraction. The nose consists of three structural layers: the outer skin envelope, middle osteocartilaginous framework, and inner mucosal lining. Many methods have been proposed to correct short nose deformity. Those procedures lengthen the nose slightly, but none of them take into account the unique characteristics of postoperative causes and the structural concept of the nose. The procedures have resulted in only limited success. On the basis of the above clinical findings and the structural concept, we developed a surgical technique to correct postoperative short nose according to the structural layers. Our method consists of three main surgical maneuvers: (1) a gull-wing concha chondrocutaneous composite graft to supplement the deficient middle and inner layers, (2) a rib costochondral onlay graft on the dorsum to reinforce the framework, and (3) wide dissection of the outer skin envelope to cover the lengthened framework without tension. We prefer a closed surgical approach rather than an open approach to avoid too much tension on the columellar incision site and to allay patients' fear of an additional scar. From 1988 to 1998, we performed our lengthening technique on six female patients. All six patients demonstrated a significant lengthening and improved appearance postoperatively. After the lengthening procedure, the average nasolabial angle improved from 116 degrees to 104 degrees. The mean follow-up period was 8.7 months, with a range of 3 to 17 months. Sometimes, epidermal sloughing in the vertical strut of the gull wing PMID- 10839422 TI - Breast augmentation: choosing the optimal incision, implant, and pocket plane. AB - A retrospective study of 220 patients was performed to review surgical design in breast augmentation. Three specific issues were studied: incision site, implant variables, and pocket plane selection. The influence of these three factors on aesthetic results in both primary and secondary cases was the focus of the analysis. No attempt was made to address long-term issues such as capsular contracture or saline implant deflation rates. In 77 primary augmentation patients and 80 unilateral augmentations for symmetry in breast reconstruction, there were the following untoward results: 11 revisions for unilateral malposition, change to a different implant shape, or change to a larger implant size; four deflations of saline implants requiring replacement; and four conversions of saline to silicone gel implants. In 63 secondary cases, there were two hematomas and two infections requiring implant removal and subsequent replacement. Operative technique in breast augmentation is described, as are recommendations for each of the options associated with the three variables studied. PMID- 10839423 TI - Intraoperative injection of botulinum toxin A into orbicularis oculi muscle for the treatment of crow's feet. AB - The purpose of this investigation was to evaluate the degree of efficacy of eliminating crow's feet by means of direct injection of botulinum toxin A into orbicularis oculi muscles under direct surgical vision during either blepharoplasty or face lift operations. Eighteen patients were injected with Botox A-14 in each orbicularis oculi muscle. Dilution was obtained by adding 4 ml of preservative-free saline to 100 IU of Botox A. Doses ranged from 15 to 50 IU in each muscle, varying according to the severity of wrinkles and intensity of muscle contraction. In 10 patients (56 percent), the Botox was injected throughout the outer surface of both orbicularis oculi dissected during a face lift operation. In eight other patients (44 percent), the toxin was injected into the inner surface of both orbicularis oculi exposed during classic blepharoplasty procedures. Most authors have demonstrated that the effect produced by transcutaneous Botox lasts between 4 and 6 months; the paralysis obtained by direct muscular injection was effective for 9 months in 14 patients (78 percent) and 10 months in the other 4 patients (22 percent). Results were documented by means of preinjection and postinjection photographs, videotapes, and electromyographs. Neither local nor general adverse effects were noted. The improvement obtained in crow's feet was satisfactory to the patient and to us. The use of Botox intraoperatively permitted at the same time not only the treatment of crow's feet by paralysis of orbicularis oculi muscles but also the correction of senile changes in the lids and face by means of either blepharoplasty or face-lift operations. PMID- 10839424 TI - The Turkish delight: a pliable graft for rhinoplasty. AB - In nose surgery, carved or crushed cartilage used as a graft has some disadvantages, chiefly that it may be perceptible through the nasal skin after tissue resolution is complete. To overcome these problems and to obtain a smoother surface, the authors initiated the use of Surgicel-wrapped diced cartilage. This innovative technique has been used by the authors on 2365 patients over the past 10 years: in 165 patients with traumatic nasal deformity, in 350 patients with postrhinoplasty deformity, and in 1850 patients during primary rhinoplasty. The highlights of the surgical procedure include harvested cartilage (septal, alar, conchal, and sometimes costal) cut in pieces of 0.5 to 1 mm using a no. 11 blade. The fine-textured cartilage mass is then wrapped in one layer of Surgicel and moistened with an antibiotic (rifamycin). The graft is then molded into a cylindrical form and inserted under the dorsal nasal skin. In the lateral wall and tip of the nose, some overcorrection is performed depending on the type of deformity. When the mucosal stitching is complete, this graft can be externally molded, like plasticine, under the dorsal skin. In cases of mild-to moderate nasal depression, septal and conchal cartilages are used in the same manner to augment the nasal dorsum with consistently effective and durable results. In cases with more severe defects of the nose, costal cartilage is necessary to correct both the length of the nose and the projection of the columella. In patients with recurrent deviation of the nasal bridge, this technique provided a simple solution to the problem. After overexcision of the dorsal part of deviated septal cartilage and insertion of Surgicel-wrapped diced cartilage, a straight nose was obtained in all patients with no recurrence (follow-up of 1 to 10 years). The technique also proved to be highly effective in primary rhinoplasties to camouflage bone irregularities after hump removal in patients with thin nasal skin and/or in cases when excessive hump removal was performed. As a complication, in six patients early postoperative swelling was more than usual. In 16 patients, overcorrection was persistent owing to fibrosis, and in 11 patients resorption was excessive beyond the expected amount. A histologic evaluation was possible in 16 patients, 3, 6, and 12 months postoperatively, by removing thin slices of excess cartilage from the dorsum of the nose during touch-up surgery. This graft showed a mosaic-type alignment of graft cartilage with fibrous tissue connection among the fragments. In conclusion, this type of graft is very easy to apply, because a plasticine-like material is obtained that can be molded with the fingers, giving a smooth surface with desirable form and long-lasting results in all cases. The favorable results obtained by this technique have led the authors to use Surgicel-wrapped diced cartilage routinely in all types of rhinoplasty. PMID- 10839425 TI - Large-volume liposuction complicated by retroperitoneal hemorrhage: management principles and implications for the quality improvement process. AB - Large-volume liposuction can be associated rarely with major medical complications and death. The case of exsanguinating retroperitoneal hemorrhage that led to cardiopulmonary arrest in an obese 47-year-old woman who underwent large-volume liposuction is described. Extensive liposuction is not a minor procedure. Performance in an ambulatory setting should be monitored carefully, if it is performed at all. Reporting of adverse events associated with outpatient procedures performed by plastic surgeons should be mandated. Hemodynamic instability in the early postoperative period in an otherwise healthy patient may be due to fluid overload, lidocaine toxicity, or to hemorrhagic shock and must be recognized and treated aggressively. Guidelines for the safe practice of large volume liposuction need to be established. PMID- 10839426 TI - Aesthetic perspectives regarding physically and mentally challenged patients. AB - Cosmetic surgery was undertaken on a limited number of physically or mentally challenged patients. Motivation for surgery seemed to mimic those of average cosmetic surgery patients, although the request for surgery was more difficult for some patients in view of their physical or mental challenges. Patient and surgeon satisfaction was extremely high. In each case, expectations were realistic. Physical or mental variations should not cloud the indications for cosmetic surgery; on the contrary, it may be a wonderful way for the plastic surgeon to contribute to bettering the life of those less fortunate. PMID- 10839428 TI - Self-promotion with commotion. PMID- 10839427 TI - Dynamics in rhinoplasty. AB - Nasal dynamics were studied on 87 patients undergoing rhinoplasty of one zone or two distant nasal zones. Statistical analysis of the results revealed that reduction of the nasion area, besides setting the soft tissue back, gave the appearance of increased intercanthal distance and lengthened the nose. Reduction of the nasal bridge resulted in a wider appearance on frontal view and a cephalically rotated tip on profile. Augmentation of the bridge affected the nose reversely. Tip cephalad rotation was achieved by resecting one of the three areas: the cephalad portion of the lower lateral cartilages (affecting the rims more), the caudal septum (affecting the central portion more), and the caudal portion of the medial crura of the lower lateral cartilages (affecting the central portion only). Resection of the alar base not only narrowed the nostrils but also moved the alar rim caudally. Furthermore, it reduced tip projection when a large alar base reduction was done. Reduction of the nasal spine increased the upper lip length on profile and reduced tip projection when a large reduction took place. Significant reduction in caudal nose projection resulted in widening of the alar base. PMID- 10839429 TI - Why climb a ladder when you can take the elevator? PMID- 10839430 TI - Breast milk lidocaine levels in tumescent liposuction. PMID- 10839431 TI - Liposuction with standing technique. PMID- 10839432 TI - Importance of the pedicle length measurement in reduction mammaplasty. PMID- 10839433 TI - Refined version of the tuberous breast classification. PMID- 10839434 TI - Coagulated blood within a replaced intact silicone gel breast implant. PMID- 10839435 TI - Latissimus dorsi donor-site quilting: less is more? PMID- 10839436 TI - Witch's chin: a progressive three-step technique. PMID- 10839437 TI - The "chemical leech" revisited. PMID- 10839438 TI - Risk of transmission of agents associated with Creutzfeldt-Jakob disease and bovine spongiform encephalopathy. PMID- 10839439 TI - A pedicle-lengthening technique for abductor hallucis muscle flap. PMID- 10839440 TI - What's wrong with Dermabond? PMID- 10839441 TI - Breast augmentation: compression--a very important factor in preventing capsular contracture. PMID- 10839442 TI - Maintenance of nipple projection using auricular cartilage. PMID- 10839443 TI - Nipple reconstruction and mastectomy scars. PMID- 10839444 TI - An inexpensive and efficient method to keep the difficult anastomotic site clear in microvascular surgery. PMID- 10839445 TI - A "painful" olive. PMID- 10839446 TI - Postoperative monitoring of microvascular tissue transplants with an implantable Doppler probe. PMID- 10839447 TI - February sees an increase in carcase submissions to the SAC. PMID- 10839448 TI - Cost-effective antimicrobial drug selection for the management and control of respiratory disease in European cattle. AB - Respiratory disease in growing cattle has both animal welfare and economic implications, but the use of antimicrobial drugs to treat and control it is under public scrutiny owing to concerns that their use in food-producing animals may be detrimental to human health. This paper outlines criteria for the selection of appropriate and cost-effective drugs, based on good dinical practice and sound economic principles. It also suggests that these principles should be integrated into quality assurance schemes, and that the monitoring of antimicrobial resistance patterns among known bacterial respiratory pathogens should be improved. PMID- 10839449 TI - Gender, age and breed pattern of diagnoses for veterinary care in insured dogs in Sweden during 1996. AB - More than 200,000 dogs insured for veterinary care by one Swedish company at the beginning of 1996 were included in a retrospective cross-sectional study. Data from receipts for visits to veterinarians that were part of reimbursed veterinary care events, whose cost exceeded the deductible cost, were used to calculate the risk of cause-specific morbidity, both in total and stratified by gender, age and breed. Diagnoses were described by organic system, for example, cardiovascular, and by disease process, for example, neoplastic The most commonly affected system was the integument followed by the gastrointestinal and genital systems. The genital system was the most commonly affected system in females, within which 50 per cent of the affected bitches had pyometra. Inflammatory processes were most common, followed by symptomatic conditions, traumatic injuries and neoplastic conditions. PMID- 10839450 TI - Diagnosis of atrophic rhinitis in a Simmental heifer by computed tomography. AB - Atrophic rhinitis was diagnosed in a 10-month-old heifer with the assistance of computed tomography. The aetiology of the condition in cattle is uncertain and the clinical disease may be overlooked owing to the lack of signs such as the shortening or deformity of the snout observed in pigs. PMID- 10839451 TI - Breed predisposition to ureteral ectopia in bitches in the UK. PMID- 10839452 TI - Clinical research and antimicrobial resistance. PMID- 10839453 TI - Retired members of the RCVS. PMID- 10839454 TI - Unseasonal snake bites in dogs. PMID- 10839455 TI - Structural heterogeneity in intramolecular DNA triple helices. AB - Oligodeoxynucleotides designed to form intramolecular triple helices are widely used as model systems in thermodynamic and structural studies. We now report results from UV, Raman and NMR experiments demonstrating that the strand polarity, which also determines the orientation of the connecting loops, has a considerable impact on the formation and stability of pyr x pur x pyr triple helices. There are two types of monomolecular triplexes that can be defined by the location of their purine tract at either the 5'- or 3'-end of the sequence. We have examined four pairs of oligonucleotides with the same base composition but with reversed polarity that can fold into intramolecular triple helices with seven base triplets and two T4 loops under appropriate conditions. UV spectroscopic monitoring of thermal denaturation indicates a consistently higher thermal stability for the 5'-sequences at pH 5.0 in the absence of Mg2+ ions. Raman spectra provide evidence for the formation of triple helices at pH 5 for oligomers with purine tracts located at either the 5'- or 3'-end of the sequence. However, NMR measurements reveal considerable differences in the secondary structures formed by the two types of oligonucleotides. Thus, at acidic pH significant structural heterogeneity is observed for the 3'-sequences. Employing selectively 15N-labeled oligomers, NMR experiments indicate a folding pattern for the competing structures that at least partially changes both Hoogsteen and Watson-Crick base-base interactions. PMID- 10839456 TI - Interdependence between a 55 kDa protein (p55) and a 12 kDa protein (p12) in facilitating the nuclear entry of goat uterine estrogen receptor under cell-free conditions. AB - A 55 kDa nuclear localization signal binding protein (p55) is involved in the transport of the goat uterine estrogen receptor from the cytoplasm to the nuclear pore complex (NPC). p55 forms a complex with a 12 kDa protein (p12) which in turn becomes 'docked' at the NPC. The present study reports on the purification and functional characterization of p12. Both p55 and p12 are Mg2+-dependent ATPases. The protein-protein interactions that take place between these two molecules at the NPC cause an enhancement in the net ATPase activity associated with the protein complex. Presumably, this enhanced ATPase function helps in the final nuclear entry of the estrogen receptor; p55 remains associated with p12 at the nuclear entry site under these conditions. PMID- 10839457 TI - Guanosine nucleotides regulate B2 kinin receptor affinity of agonists but not of antagonists: discussion of a model proposing receptor precoupling to G protein. AB - The effect of nucleotides on binding of the B2 kinin (BK) receptor agonist [3H]BK and the antagonist [3H]NPC17731 to particulate fractions of human foreskin fibroblasts was studied. At 0 degrees C, particulate fractions exhibited a single class of binding sites with a Kd of 2.3 nM for [3H]BK and a Kd of 3.8 nM for the antagonist [3H]NPC17731. Incubation with radioligands at 37 degrees C for 5 min gave a reduction of agonist, as well as antagonist, binding that was between 0 40% depending on the preparation, even in the absence of guanosine nucleotides. As shown by Scatchard analysis, this reduction in specific binding was due to a shift in the affinity of at least a fraction of the receptors. The presence at 37 degrees C of the guanine nucleotides GTP, GDP and their poorly hydrolyzable analogs left [3H]NPC17731 binding unaffected, but reduced the receptor affinity for [3H]BK to a Kd of about 15 nM. The maximal number of receptors, however, was unchanged. This affinity change was strongly dependent on the presence of bivalent cations, in particular Mg2+. It was reversed by incubation at 0 degrees C. The rank order of the guanosine nucleotides for [3H]BK binding reduction was GTP[gammaS] = Gpp[NH]p > GTP = GDP > GDP[betaS]. GMP, ATP, ADP and AMP showed no influence on agonist binding. A model for the interaction of the B2 kinin receptor with G proteins is discussed. PMID- 10839458 TI - Regulation of PTHrP and PTH/PTHrP receptor by extracellular Ca2+ concentration and hormones in the breast cancer cell line 8701-BC. AB - It was previously reported that 8701-BC breast tumour cells express the gene for parathyroid hormone-related peptide (PTHrP) and PTH/PTHrP receptor (PTHrP-R) and release immunoreactive PTHrP (iPTHrP) into the extracellular medium. Since the regulation of PTHrP and PTHrP-R by breast cancer cells has been poorly investigated so far, we have chosen the 8701-BC cell line as a model system to investigate whether alterations in the extracellular Ca2+ concentration ([Ca2+]e) and treatment with some well-known differentiation agents for breast cells, such as dimethyl sulfoxide, hydrocortisone, progesterone, prolactin, all-trans retinoic acid and transforming growth factor-beta1 might (i) modulate quantitatively the release of iPTHrP, (ii) affect the PTHrP promoter usage and mRNA splicing patterns, and (iii) modify the expression of PTHrP-R. The data obtained indicate that 8701-BC cells are potentially able to utilise different start sites and mRNA splicing patterns for PTHrP transcription, and respond to variations of [Ca2+]e and to the addition of two hormones, hydrocortisone and progesterone, with modifications in the extracellular amount of iPTHrP. Moreover, expression of PTHrP-R is also modulated by changes of [Ca2+]e or treatment with hydrocortisone. This indicates that the 8701 -BC cell line is a suitable in vitro model for further studies on the complex molecular regulation of the PTHrP/PTHrP R pair in breast cancer. PMID- 10839459 TI - Effect of vanadyl ions on calcineurin and its A subunit. AB - Calcineurin (CN) exhibits a bimodal regulation by different concentrations of vanadyl ions (VO2+) in the presence of Mn2+. Low concentrations of VO2+ inhibit the enzyme, with 50 microM VO2+ completely inhibiting CN activity, while high concentrations, up to 500 microM VO2+, stimulate the CN activity. A similar bimodal regulation of CN was not observed with either calcium or vanadate under the same conditions. X-band electron spin resonance spectroscopy, used to study the binding of VO2+ to the catalytic subunit A of calcineurin, show that there are two kinds of binding sites in the A subunit. PMID- 10839460 TI - Action mechanism of tyrosinase on meta- and para-hydroxylated monophenols. AB - The relationship between the structure and activity of meta- and para hydroxylated monophenols was studied during their tyrosinase-catalysed hydroxylation and the rate-limiting steps of the reaction mechanism were identified. The para-hydroxylated substrates permit us to study the effect of a substituent (R) in the carbon-1 position (C-1) of the benzene ring on the nucleophilic attack step, while the meta group permits a similar study of the effect on the electrophilic attack step. Substrates with a -OCH3 group on C-1, as p-hydroxyanisol (4HA) and m-hydroxyanisol (3HA), or with a -CH2OH group, as p hydroxybenzylalcohol (4HBA) and m-hydroxybenzylalcohol (3HBA), were used because the effect of the substituent (R) size was assumed to be similar. However, the electron-donating effect of the -OCH3 group means that the carbon-4 position (C 4) is favoured for nucleophilic attack (para-hydroxylated substrates) or for electrophilic attack (meta-hydroxylated substrates). The electron-attracting effect of the -CH2OH group has the opposite effect, hindering nucleophilic (para) or electrophilic (meta) attack of C-4. The experimental data point to differences between the maximum steady-state rate (V(M)Max) of the different substrates, the value of this parameter depends on the nucleophilic and electrophilic attack. However, differences are greatest in the Michaelis constants (K(M)m), with the meta-hydroxylated substrates having very large values. The catalytic efficiency k(M)cat/K(M)m is much greater for thepara-hydroxylated substrates although it varies greatly between one substrate and the other. However, it varies much less in the meta-hydroxylated substrates since this parameter describes the power of the nucleophilic attack, which is weaker in the meta OH. The large increase in the K(M)m of the meta-hydroxylated substrates might suggest that the phenolic OH takes part in substrate binding. Since this is a weaker nucleophil than the para hydroxylated substrates, the binding constant decreases, leading to an increase in K(M)m. The catalytic efficiency of tyrosinase on a monophenol (para or meta) is directly related to the nucleophilic power of the oxygen of the phenolic OH. The oxidation step is not limiting since if this were the case, the para and meta substrates would have the same V(M)max. The small difference between the absolute values of V(M)max suggests that the rate constants of the nucleophilic and electrophilic attacks are on the same order of magnitude. PMID- 10839461 TI - Urethanyl-3-amidinophenylalanine derivatives as inhibitors of factor Xa. X-ray crystal structure of a trypsin/inhibitor complex and modeling studies. AB - Hydrophobic urethanyl derivatives of 3-amidinophenylalanine methyl ester were found to be relatively potent and selective factor Xa inhibitors. These compounds consist of the arginine-mimetic 3-benzamidino group as P1 residue and of hydrophobic residues as potential interaction partners for the S3/S4 aryl binding site of the enzyme. Attempts to possibly identify their binding mode to factor Xa via the X-ray crystal structure of a trypsin/inhibitor complex and analogy modeling on the crystal structure of factor Xa failed. However, synthesis of enantiomerically pure (R)- and (S)-derivatives, combined with modeling experiments, led to an hypothetical non-substrate like binding mode, which was fully confirmed by the remarkably enhanced inhibitory potency of derivatives in which the methyl ester was replaced by arylamides for interactions with the S3/S4 enzyme binding subsites. With adamantyloxycarbonyl-(R)-3-amidinophenylalanine phenethylamide+ ++ a nanomolar inhibiton was obtained, thus indicating this new class of factor Xa inhibitors as a highly promising lead structure. PMID- 10839462 TI - Cloning and characterization of the human PAX7 promoter. AB - PAX7, a member of the PAX transcription factor gene family, is normally expressed at high levels during development in the neural tube and in skeletal muscle precursor cells. Interestingly, PAX7 expression was also identified in tumor cells developing from these cell types. To date not much is known about the molecular mechanisms controlling the regulation of PAX7 expression. Therefore, we have cloned and sequenced part of the proximal 5'-flanking region of the human PAX7 gene. Computer-based sequence analysis identified putative binding sites for basic transcription factors. Analysis of a series of deletion constructs in different cell types suggested that a distal region containing several E-boxes might be involved in muscle-specific expression of PAX7, and that a distinct proximal region can enhance basal PAX7 expression in tumor cells. PMID- 10839463 TI - Genomic organization, expression analysis, and chromosomal localization of the mouse PEX3 gene encoding a peroxisomal assembly protein. AB - The peroxin Pex3p has been identified as an integral peroxisomal membrane protein in yeast where pex3 mutants lack peroxisomal remnant structures. Although not proven in higher organisms, a role of this gene in the early peroxisome biogenesis is suggested. We report here the cDNA cloning and the genomic structure of the mouse PEX3 gene. The 2 kb cDNA encodes a polypeptide of 372 amino acids (42 kDa). The gene spans a region of 30 kb, contains 12 exons and 11 introns and is located on band A of chromosome 10. The putative promoter region exhibits characteristic housekeeping features. PEX3 expression was identified in all tissues analyzed, with the strongest signals in liver and in testis, and could not be induced by fenofibrate. The data presented may be useful for the generation of a mouse model defective in PEX3 in order to clarify the yet unknown functional impact of disturbances in early peroxisomal membrane assembly. PMID- 10839464 TI - Expression and functional characterization of a pHis-tagged human bradykinin B2 receptor in COS-7 cells. AB - A polyHis-tagged bradykinin (BK) B2 receptor (pHis-BKR) cDNA was constructed and expressed in COS-7 cells. The pHis-BKR is suitable for both immunoprecipitation and immunoblotting with anti-polyHis antibodies and can be easily purified using Ni-NTA columns. Immunochemical detection revealed a molecular mass of approximately 66 kDa. The pHis-BKR is capable of mediating BK-induced stimulation of inositol phosphate formation as well as of mitogen-activated protein kinase (MAPK) activity. Compared with the wild-type receptor (WT-BKR) the tagged receptor showed a slightly enhanced affinity towards BK but a reduced expression level. Despite these modified pharmacological properties the pHis-tagged BKR may be a useful tool for studying BKR modifications and signaling. PMID- 10839465 TI - Purification, characterisation and cDNA sequencing of pyruvate decarboxylase from Zygosaccharomyces bisporus. AB - Cells of the wild-type yeast strain Zygosaccharomyces bisporus CBS 702 form alpha hydroxy ketones from aromatic amino acid precursors during fermentation. Pyruvate decarboxylase (PDC, E.C. 4.1.1.1), the key enzyme of this biotransformation catalysing the non-oxidative decarboxylation of pyruvate and other 2-oxo-acids, was purified and characterised. The active enzyme is homotetrameric (alpha4) with a molecular mass of about 244 kDa. Activation of PDC by its substrate pyruvate results in a sigmoidal dependence of the reaction rate from substrate concentration (apparent Km value 1.73 mM; Hill coefficient 2.10). A cDNA library was screened using a PCR-based procedure, and a 1856 bp cDNA of PDC was identified and sequenced. The cDNA encodes a polypeptide of 563 amino acid residues (monomeric unit). Sequence alignments demonstrate high homologies (> 80%) to PDC genes from Saccharomyces cerevisiae, Kluyveromyces lactis and Kluyveromyces marxianus. PMID- 10839466 TI - Trospium chloride: an effective option for medical treatment of bladder overactivity. PMID- 10839467 TI - Variation of CYP1A2-dependent caffeine metabolism during menstrual cycle in healthy women. AB - BACKGROUND AND OBJECTIVES: The activity of the human cytochrome P450 CYP1A2 is decreased by female sex hormones during pregnancy or treatment with oral contraceptives. However, the influence of menstrual cycle on CYP 1A2 activity is not clear. METHODS: CYP1A2 activity was monitored in 15 women (13 with confirmed ovulatory cycles, 2 smokers, age (mean +/- SD) 27.8 +/- 3.8 years, body mass index 23.8 +/- 3.8 kg x m-2) using the specific substrate caffeine (mean doses 149 mg). After a run-in period started one week prior to expected onset of menses, daily saliva samples were taken 7.3 +/- 0.7 hours after caffeine intake throughout the cycle, and caffeine clearance was estimated from the paraxanthine to caffeine ratio therein. Ovulation was confirmed by progesterone serum concentration above 3 ng/ml in the second half of the cycle. RESULTS: Initial (day 2) caffeine clearance (n = 15, geometric mean) was 1.37 ml/min/kg body weight (coefficient of variation (CV) 48%). The ratio of caffeine clearance for the luteal (day -9 to -4 prior to onset of the next menses) to the follicular phase (days 5-10) was (n = 13, point estimate) 1.03 (90% CI 0.95-1.12), indicating that there was no difference in CYP1A2 activity between these cycle phases. The median intraindividual CV in ovulatory cycles (n = 13) was 23% (range 11% to 39%). As an additional finding, there was evidence for long-term fluctuations of CYP1A2 activity in most individuals. CONCLUSIONS: A dose adaptation according to the phase of menstrual cycle based on pharmacokinetics is not required for CYP1A2 substrates. PMID- 10839468 TI - Effect of chronic renal failure on the disposition of highly hepatically metabolized drugs. AB - OBJECTIVE: The objective of this study was to investigate the effect of renal impairment on the disposition of an extensively metabolized drug, i.e., drug X. Drug X has a hepatic extraction ratio of less than 0.1 and free fraction in plasma of less than 1% in healthy volunteers. METHODS: Pharmacokinetic (PK) parameters of drug X were obtained from subjects with normal renal function (I, n = 6), as well as in subjects with mild (II, n = 5), moderate (III, n = 7) and severe renal impairment (IV, n = 5). Disease-PK models were developed to describe the changes of PK parameters with respect to renal function measured by creatinine clearance. While experimentally observed data are presented for drug X, additional simulations were performed for other drugs that are extensively metabolized (extensive metabolism is defined as metabolism that accounts for more than 90% of total drug elimination). The simulated scenarios included drugs that have a low extraction ratio (ER) and with high plasma protein binding (PPB), low ER and with low PPB, high ER and with high PPB, or high ER and with low PPB. RESULTS: Systemic clearance of drug X, a low ER and high PPB drug, in renal patients depended on the simultaneous effects of renal disease on protein binding and intrinsic metabolic clearance. Protein binding of drug X was related to creatinine clearance in an inverse hyperbolic relationship, while the unbound intrinsic metabolic clearance declined linearly with creatinine clearance. Because the disease effects on these two factors offset each other in terms of total systemic clearance, the lowest total systemic clearance was not observed in the severely renal impairment patients, but rather in the moderately impaired group. Additional simulations showed that for low ER drugs that are highly metabolized, the pattern and magnitude of systemic clearance change in renal patients depended on how the disease affected PPB and/or intrinsic metabolic clearance. But the systemic clearance of high ER drugs would not be as susceptible to the effect of renal disease as that of low ER drug. CONCLUSIONS: Chronic renal disease should not be considered as an isolated event that affects only renally excreted drugs. Uremia may also modify the disposition of a highly metabolized drug by changes in plasma protein binding and/or hepatic metabolism. PMID- 10839469 TI - Pharmacokinetics of reboxetine in elderly patients with depressive disorders. AB - OBJECTIVES: To examine the pharmacokinetic characteristics of the selective norepinephrine reuptake inhibitor, reboxetine, in elderly patients with depression. PATIENTS: Twelve female inpatients (mean age 80 +/- 4 years) with major depressive or dysthymic disorder were enrolled in a 4-week uncontrolled study of oral reboxetine 2-8 mg/day. METHODS: After a one-week washout period, patients were randomized into two groups (groups A and B, n = 6/group). Reboxetine was given twice daily, starting with 2 mg/day during week 1 and increasing by 2 mg/day each week to 8 mg/day in week 4. Pharmacokinetic evaluations were carried out at two dosage levels in each group: at the end of weeks 1 and 3 in group A (2 and 6 mg/day), and at the end of weeks 2 and 4 in group B (4 and 8 mg/day). Blood and urine samples were taken for determination of reboxetine pharmacokinetics. RESULTS: Reboxetine displayed linear pharmacokinetics, with dose-proportional changes, in elderly depressed patients. Mean total urinary recovery ranged from 4.06 to 6.17%. The mean area under the plasma concentration-time curve (AUCtau) and the maximum plasma drug concentration (Cmax) showed considerable variation between patients; at a dosage of 4 mg/day, AUCtau was 1,466-6,866 ngxh/ml and Cmax ranged from 169 to 663 ng/ml. CONCLUSIONS: The pharmacokinetics of reboxetine are linear across the dosage range of 2-8 mg/day in elderly depressed patients, although Cmax and AUCtau values are higher (and more variable) than in young adults. These results support the use of a lower starting dose (4 mg/day) of reboxetine in the elderly. PMID- 10839470 TI - Ethnic differences in response to non-selective beta-blockade among racial groups in Malaysia. AB - OBJECTIVE: To determine whether racial differences in response to blockade of beta receptors occur among racial groups in Malaysia that are the Malays, Indians and Chinese. SUBJECTS, MATERIALS AND METHOD: 35 healthy male volunteers representing the 3 main racial groups in Malaysia (12 Malays, 12 Chinese and 11 Indians) were studied in a randomized, placebo-controlled, crossover and single blind design. Propranolol 80 mg 12-hourly was given orally for 48 hours. Six hours after the last dose subjects attended an exercise session where resting and exercise heart rate, blood pressure, plasma potassium and glucose levels, resting FEV1 and plasma propranolol concentrations were recorded. RESULTS: No significant difference in plasma propranolol (mean +/- SEM) levels was seen between races six hours after the last dose (Malays, 59.7 +/- 8.8 ng/ml, Indians, 67.6 +/- 19.3 ng/ml, Chinese, 58.4 +/- 7.9 ng/ml). Chinese were least sensitive to the bradycardic and hypotensive effects of propranolol at rest and exercise. Indians and Malays had significant reduction of supine systolic blood pressure with propranolol but not Chinese. Comparison of percentage reductions of systolic blood pressure at supine, sitting and exercise by repeated measure analysis showed the Malays to have significantly higher change compared to the Chinese (p = 0.022). Similarly, comparison of percentage reductions of heart rate at supine, sitting and exercise by repeated measure analysis showed the Malays to have significantly higher change compared to the Chinese (p = 0.040). Average change in potassium concentrations at peak exercise and recovery showed the Indians to have significantly higher increase in potassium levels with propranolol compared to the Malays (p = 0.038). However, no significant interethnic difference was seen in the reduction of glucose levels at rest, peak exercise or recovery. Also, no significant interethnic difference was seen in reduction of FEV1 values. CONCLUSION: We, therefore, conclude that ethnic differences in response to blockade of beta-receptors exist among racial groups in Malaysia. These differences were seen at similar plasma drug levels between races suggesting ethnic differences in drug sensitivity, rather than differences in drug disposition. PMID- 10839471 TI - Fluvastatin increases prostacyclin and decreases endothelin production by human umbilical vein endothelial cells. AB - Fluvastatin, an agent of a class of lipid-lowering drugs, the "statins", significantly enhanced prostacyclin synthesis at the concentrations of 0.1 microM and 1 microM and significantly reduced endothelin production at the concentrations 0.01, 0.1 and 1 microM in cell cultures of human umbilical endothelial veins. Since prostacyclin is a vasodilator and endothelin a vasoconstrictor, fluvastatin may have a significant effect on hemodynamics by favoring the balance towards vasodilation. This mechanism may contribute to the prevention of cardiovascular diseases. PMID- 10839472 TI - The carcinogenicity of the biocide ortho-phenylphenol. AB - The biocides ortho-phenylphenol and its sodium salt (OPP and SOPP) are widely used as fungicides and antibacterial agents for commercial and consumer purposes. The carcinogenicity of OPP/SOPP toward the urinary bladder was demonstrated when rats were chronically fed concentrations of 0.5%-4% in their diet. Other species tested so far did not develop tumours. Understanding the mechanisms underlying OPP/SOPP-induced bladder carcinogenesis is critical to determine whether risks observed at high doses in rats are of relevance to humans exposed at much lower levels. This overview details experimental studies of carcinogenicity, genotoxicity as well as metabolism/toxicokinetics and other mechanistic studies which bear on cancer hazard and risk evaluation of exposure to humans. Based on the presently available knowledge, it is concluded that reactive quinoid metabolites exhibiting redox cycling activities are the crucial factors. At certain concentration levels, these metabolites are able to produce cytotoxic events with concomitant enhanced cell proliferation of the target tissue. Further important risk factors are probably promutagenic lesions induced by oxidative stress and a higher urinary pH. Supposed that these mechanisms are the basis for the tumourigenicity observed, then suitable low doses of OPP/SOPP will practically pose no cancer risk. PMID- 10839473 TI - New methods for determination of 2-butoxyethanol, butoxyacetaldehyde and butoxyacetic acid in aqueous systems, with special reference to cell culture conditions. AB - Ethylene glycol ethers, especially 2-ethoxyethanol and 2-butoxyethanol (BE) are frequently used in industry and household as solvents and detergents because of their excellent hydrophilic and lipophilic properties. BE and its oxidation products, butoxyacetaldehyde (BAL) and butoxyacetic acid (BAA), are mainly associated with haemolytic toxicity. No method to determine BAL in aqueous systems (e.g. urine or blood) has been published up to now. BAL was synthesized by dehydration of BE and identified by gas chromatography-mass spectrometry. For determination of BAL and BE with head space-capillary gas chromatography, water and HCl or sodium dihydrogen phosphate were added to the sample. No further extraction or derivatization were necessary. For BAA determination after adding HCl and sodium dihydrogen phosphate the samples were extracted with ethyl acetate and derivatized with 2,2,2-trichloroethanol/HCl. The analytical methods presented here are reliable, sensitive and rapid. The new methods were developed for mammalian cell culture systems, because such in vitro systems are especially useful for metabolic studies and have the advantage of choosing species and organ specificity. In the cell culture experiments presented here it was demonstrated that Opossum kidney cells are able to metabolize BAL to BAA within 24 h. After this interval, in the cells neither BAL nor BAA were accumulated, whereas BAA was found in the cell culture media. PMID- 10839474 TI - Combined effects of okadaic acid and cadmium on lipid peroxidation and DNA bases modifications (m5dC and 8-(OH)-dG) in Caco-2 cells. AB - Okadaic acid (OA) is a marine toxin, a tumour promoter and an inducer of apoptosis. It mainly inhibits protein-phosphatases, protein synthesis and enhances lipid peroxidation. Cadmium (Cd) is known to be carcinogenic in animals and humans (group 1 according to the International Agency for Research on Cancer (IARC) classification). Cd also induces oxidative stress in living organisms. Since they are sometimes found simultaneously in mussels, we have evaluated in the present investigation, the lipid peroxidation, as malondialdehyde (MDA) production, in the variation of the ratios of 8-(OH)-dG/10(5)dG and m5dC/(dC + m5dC) induced by OA and/or Cd in Caco-2 cells. When cells were treated exclusively by OA (15 ng/ml) or Cd (0.625 and 5 microg/ml) for 24 h, protein synthesis was inhibited (by 42 +/- 5%, 18 +/- 13%, and 90 +/- 4% respectively) while MDA production was 2,235 +/- 129, 1710 +/- 20, and 11,496 +/-1,624 pmol/mg protein respectively. In addition, each toxicant induced modified bases in DNA; increases in oxidised bases and methylated dC. The combination of OA and cadmium was more cytotoxic and caused more DNA base modifications; the ratio m(5)dC/(m(5)dC + dC) was increased from 3 +/- 0.15 to 9 +/- 0.15 and the ratio 8 (OH)-dG/10(5) dG also (from 36 +/- 2 to 76 +/- 6). The combination of OA and Cd also increased the level of MDA (1,6874 +/- 2,189 pmole/mg protein). The present results strongly suggest that DNA damage resulting from the oxidative stress induced by these two toxicants may significantly contribute to increasing their carcinogenicity via epigenetic processes. PMID- 10839475 TI - Species differences in response to diethylhexylphthalate: suppression of apoptosis, induction of DNA synthesis and peroxisome proliferator activated receptor alpha-mediated gene expression. AB - Diethylhexylphthalate (DEHP) is a phthalate plasticizer that belongs to the peroxisome proliferator (PP) class of rodent nongenotoxic hepatocarcinogens. Previously, we have shown that MEHP (a principal metabolite of DEHP and the proximal PP) induced DNA synthesis and suppressed apoptosis in rat but not in human hepatocytes in vitro. Here, we present further studies of species differences in response to DEHP. In rats, 4 days of exposure to DEHP (950 mg/kg per day by gavage) induced peroxisomal beta-oxidation, DNA synthesis and suppressed apoptosis. In contrast, there was no response of guinea pig liver to DEHP. In rat hepatocytes in vitro, MEHP (250, 500 and 750 microM) induced peroxisomal beta-oxidation, DNA synthesis and suppressed apoptosis. In contrast to the pleiotropic response noted in rat hepatocytes, there was no response of human hepatocytes to 250, 500 or 750 microM MEHP. PPs activate the peroxisome proliferator activated receptor alpha (PPARalpha) that binds to DNA at peroxisome proliferator response elements (PPREs) within the promoters of PP-responsive genes such as rat acyl CoA oxidase (ACO). However, the human ACO gene promoter differs at three bases within the PPRE from the rat ACO promoter and appears refractory to PPs. To address species differences in response to DEHP at the molecular level, we used promoter-reporter gene assays to compare the ability of MEHP to induce gene expression from the rat or the human ACO promoter. MEHP gave a concentration-dependent increase in reporter gene expression from the rat ACO gene promoter with either mouse or human PPARalpha. In contrast, the human ACO promoter was unable to drive MEHP-induced gene transcription irrespective of the species origin of PPARalpha. These data provide further weight of evidence at the cellular and molecular levels for a lack of risk to human health from the phthalate DEHP. PMID- 10839476 TI - Effects of xenoestrogen bisphenol A on uterine and pituitary weight, serum prolactin levels and immunoreactive prolactin cells in ovariectomized Wistar rats. AB - Considerable attention has currently been focused on bisphenol A (BPA), an environmental endocrine disrupting chemical that has oestrogenic activity. In vitro and in vivo short-term assays have shown that BPA is weakly estrogenic. In addition, the issue of species- and strain-differences in susceptibility to BPA was raised. The treatment of ovariectomized (OVX) Wistar rats with BPA at doses of 11-250 mg/kg per day, s.c., for 7 days, resulted in significant dose-dependent regrowth of uterus in uterotrophic assay. Additionally, the stimulation of anterior pituitary gland growth and induction of hyperprolactinaemia, as determined by wet organ weight and radioimmunoassay (RIA), respectively, were also dose-dependent (at 128 and 250 mg/kg per day, P < 0.05). Prolactin immunostaining of anterior pituitary glands revealed that BPA at a dose of 250 mg/kg per day increased the number of prolactin-immunopositive cells by 63% compared to OVX rats. These results demonstrate that the reproductive tract and neuroendocrine axis of Wistar rats are able to respond to BPA. Furthermore, the pituitary gland hypertrophy and hyperprolactinaemia can be mediated, at least partly, by increase in number of prolactin-immunoreactive cells. The long-term consequences of this proliferation are yet unknown but neoplasm formation is an obvious possibility. PMID- 10839478 TI - Inhibitory effects of subcutaneous dexamethasone treatment on rat pulmonary toxicity of KW-2149, a new mitomycin C analogue. AB - An experimental model for pulmonary toxicity of KW-2149, a new mitomycin C analogue, was established and the inhibitory effects of dexamethasone (DM) were investigated. KW-2149 was given to male rats 3 or 5 times at weekly intervals by intravenous injection of 3.28 or 8.2 mg/kg. As a suitable model for pulmonary toxicity, the dose of 3.28 mg/kg per week for 3 weeks was selected, this causing exudative pleural effusion in all animals but no deaths. For preventing this toxicity, DM was injected subcutaneously 3 times every week at 0.5, 1.0, 2.5 or 5.0 mg/kg. The 0.5 mg/kg dose was sufficient to completely prevent development of pleural effusions. Combined DM treatment may be an effective chemotherapy for KW 2149 induced pulmonary toxicity. PMID- 10839477 TI - Metabolism and cytotoxicity of bisphenol A and other bisphenols in isolated rat hepatocytes. AB - The relation between the metabolism and the cytotoxic effects of bisphenol A (BPA, 2,2-bis(4-hydroxyphenyl)propane) has been studied in freshly isolated rat hepatocytes and isolated hepatic mitochondria. The incubation of hepatocytes with BPA (0.25-1.0 mM) elicited a concentration- and time-dependent cell death, accompanied by losses of intracellular ATP and total adenine nucleotide pools. BPA at a low-toxic level (0.25 mM) in the hepatocyte suspensions was rapidly converted to its major conjugate, BPA-glucuronide, and other minor products without marked loss of cell viability, although at a toxic level (0.5 mM), more than 65% of the compound presented in an unaltered form 2 h after the incubation. Addition of salicylamide (2 mM), non-toxic to hepatocytes during the incubation period, enhanced BPA-induced cytotoxicity and reduced the loss of BPA and the formation of BPA-glucuronide. The addition of BPA to isolated hepatic mitochondria caused a concentration (0-0.5 mM)-dependent increase in the rate of state 4 oxygen consumption in the presence of an FAD-linked substrate (succinate), indicating an uncoupling effect, whereas the rate of state 3 oxygen consumption was inhibited by BPA. Further, the addition of BPA (0.25 mM) reduced state 3 respiration with NAD+-linked substrates (pyruvate plus malate) and/or with the FAD-linked substrate, whereas state 3 respiration with ascorbate plus tetramethyl-p-phenylenediamine (cytochrome oxidase-linked respiration) was not significantly affected by BPA. A comparative study of the toxic effects of BPA and some bisphenols on cell viability (at 1.0 mM) and mitochondrial respiration (at 0.25 mM) revealed that 4,4'-(1,2-diethyl-1,2-ethenediyl)bisphenol (diethylstilbestrol) was more toxic than BPA, followed by 4,4'-methylenediphenol and 4,4'-biphenol. These results indicate that the onset of cytotoxicity caused by BPA may depend on the intracellular energy status and that mitochondria are important targets of the compound. The toxicity caused by the inhibition of ATP synthesis may be related to the concentration of unmetabolised free BPA remaining in the cell suspensions. In addition, the toxic potency of bisphenols to hepatocytes and mitochondria depends on the relative elongation and/or molecular size of the hydrocarbon bridge between the phenolic groups. PMID- 10839479 TI - Prevention by vitamin E of DNA fragmentation and apoptosis induced by fumonisin B1 in C6 glioma cells. AB - Fumonisin B1 (FB1), produced by the fungus Fusarium moniliforme, belongs to a class of sphingosine analogue mycotoxins that occur widely in the food chain. Epidemiological studies have associated consumption of Fusarium moniliforme contaminated food with human oesophageal cancer in China and South Africa. FB1 also causes equine leucoencephalomalacia. Evidence for induction of apoptosis by FB1 was first obtained when C6 glioma cells were incubated with fumonisin B1 (3 27 microM) causing DNA fragmentation profiles showing DNA laddering in gel electrophoresis and apoptotic bodies revealed by chromatin staining with acridine orange and ethidium bromide. Further confirmation experiments and comet assays have been performed under similar conditions. The results of the comet test show that FB1 at 9 and 18 microM induces respectively 50 +/- 2% and 40 +/- 1% of cells with a comet with an increased tail length of 93 +/- 9 microm and 102 +/- 17 microm respectively. Under these concentrations, FB1 induced DNA fragmentation and laddering and many apoptotic bodies. Pre-incubation of the cells with vitamin E (25 microM) for 24 h before FB1 (18 microM) significantly reduced DNA fragmentation and apoptotic bodies induced by FB1. PMID- 10839480 TI - Embryo lethality and teratogenicity of a new fluoroquinolone antibacterial DW-116 in rats. AB - DW-116, 1-(5-fluoro-2-pyridyl)-6-fluoro-7-(4-methyl-l-piperazinyl)-1, 4-dihydro-4 oxoquinolone-3-carboxylic acid hydrochloride, is a newly developed fluoroquinolone antibacterial. The potential of DW-116 to induce developmental toxicity was, investigated in Sprague-Dawley rats. DW-116 was administered by gavage to pregnant rats from days 6 to 16 of gestation at dose levels of 0, 31.3, 125, and 500 mg/kg per day. All dams were subjected to caesarean section on day 20 of gestation and their fetuses were examined for external, visceral and skeletal abnormalities. At 500 mg/kg, toxic effects including clinical signs of toxicity, suppressed body weight and decreased food intake were found in dams. An increase in the resorption rate, a decrease in the litter size, a reduction in the fetal weight, and a decrease in the placental weight were also seen. In addition, various types of external, visceral, and skeletal malformations occurred at an incidence of 17.9, 74.2 and 8.3%, respectively. Characteristic malformations included oedema, cleft palate, dilated cerebral ventricle, hypoplasia of lung and ventricular septum defect. A dramatic increase in the incidence of skeletal variations (55.6%) and retardations (94.4%) and a decrease in the number of ossification centres of sternebra, metacarpals, metatarsals and sacrocaudal vertebra were also observed. At 125 mg/kg, a reduction in the placental weight and an increase in the incidence of skeletal variations were found. There were no signs of maternal toxicity or embryotoxicity at 31.3 mg/kg. These results indicate that the fluoroquinolone antibacterial DW-116 is embryotoxic and teratogenic at minimally maternally toxic dose and is minimally embryotoxic at nonmaternally toxic dose in rats. PMID- 10839481 TI - Familial idiopathic facial palsy. AB - The etiology of idiopathic facial palsy (IFP) is still not defined. Familial inheritance has been found in 4-14% of cases. Among 116 patients with IFP admitted to the otolaryngology and neurology clinics at Kutahya State Hospital, 12 had positive family histories involving 10 different families. Review of the patients' histories showed that 13 members of families previously had IFP. In all, the 25 patients with familial IFP consisted of 12 males and 13 females and had an average age at onset of 34.3 years. Class I human leukocyte antigen (HLA) was investigated in 11 patients with familial IFP belonging to four separate families. The follow-up period was approximately 2 months. All cases except one recovered completely, with the latter patient having minimal contracture and facial paresis. However, HLA showed no significant increases in class I antigens. No notable difference was found in the clinical courses of patients with familial and non-familial IFP. PMID- 10839482 TI - Detection of human papillomavirus in cholesteatomas. AB - The presence of human papillomavirus DNA in cholesteatoma may have some role in the development of middle ear cholesteatoma as well as in papilloma. In the present study, polymerase chain reaction and in situ hybridization with human papillomavirus (HPV)-6 and -11 DNA probes were used to detect the presence of HPV DNA in 32 human middle ear cholesteatomas. Only one specimen contained HPV-6 DNA. Although its occurrence may have been coincidental, it is also possible that the hyperproliferative epithelium of cholesteatomas might have some relationship with HPV infections. PMID- 10839483 TI - Hearing loss after cardiopulmonary bypass surgery. AB - Hearing loss (HL) is a rare complication following cardiac surgery with extracorporal circulation (CSWEC) or other non-otologic surgery under general anesthesia, as is HL caused by loss of cerebrospinal fluid during neurosurgery or spinal anesthesia. The incidence of HL after CSWEC is not known since preoperative hearing testing is not commonly done and a perioperative HL may occur unnoticed. We present four cases of profound sensorineural hearing loss following CSWEC for coronary artery bypass or cardiac valve surgery. The hearing loss was noticed immediately on waking from anesthesia in three of the patients, whereas the fourth patient noticed decreased hearing during the first postoperative week. In all patients audiological investigation suggested a cochlear etiology of the HL. The possible mechanisms for HL are discussed. HL after CSWEC is most likely caused by a microembolism generated by atheromatous material. PMID- 10839484 TI - The effect of impulse noise exposure on distortion product otoacoustic emissions in the awake guinea pig. AB - Distortion product otoacoustic emissions (DPOAE) are a sensitive detector of outer hair cell (OHC) function and were monitored in awake guinea pigs before and after impulse noise damaging the cochlea (peak intensity 153 dB SPL, rise time < 0.1 ms). Animals had stable DPOAE levels before noise exposure. In the first hours after noise exposure DPOAE levels were reduced significantly. Three different patterns of recovery of DPOAE were seen in the post-exposure period:restitution exceeding controls, partial recovery and no recovery. In general, DPOAE levels declined and types of recovery closely corresponded to changes in amplitudes of cochlear microphonics after noise exposure. These data suggest that the monitoring of DPOAE is a suitable method for diagnosing impaired OHC function. PMID- 10839485 TI - The canalith repositioning maneuver in patients with benign positional vertigo. AB - The canalith repositioning maneuver (CRM), as defined by Epley, can be an effective treatment for benign paroxysmal positional vertigo (BPPV). The staff at Baskent University's Ear Nose and Throat Clinic performed CRM on 68 cases of canalithiasis in 64 BPPV patients from June 1996 to August 1997. Symptoms resolved after the first session in 49 patients (72%) and after the second session in 11 cases (16.2%). It was necessary to repeat the maneuver three times in two cases (2.9%) and four times in one patient (1.5%). Discounting three patients who were lost to follow-up, only two patients in our study did not respond to CRM treatment. There was no co-existing pathology found in all but two of the patients studied. Our experience indicates that unless there is no response to CRM or there is suspicion of an incorrect diagnosis, it is not necessary to perform diagnostic studies routinely for differentiating other neuro otologic disorders prior to using CRM in BPPV patients diagnosed by the Dix Hallpike test. PMID- 10839486 TI - HLA patterns in patients with nasal polyposis. AB - The etiology of nasal polyposis is still unknown, although risk factors include Aspirin intolerance, asthma, cystic fibrosis and primary ciliary dyskinesia. We studied frequencies of HLA A, B, DR and DQ in patients with nasal polyposis in order to determine a possible genetic component in the multifactorial pathogenesis of nasal polyps. Previous work has suggested an association of HLA A1B8 with nasal polyposis and Aspirin intolerance. We investigated 89 patients with nasal polyposis, 11 of whom had Aspirin-intolerance, 19 asthma and 22 allergies to inhalation allergens. HLA patterns of these patients were compared to the ones of 1,070 healthy controls. No significant association of HLA-A1B8 was found with nasal polyps in the Aspirin-sensitive subgroup of our patients, but a significant association was seen with HLA-A74 and nasal polyposis. PMID- 10839487 TI - Efficacy and safety of cefdinir in the treatment of maxillary sinusitis. AB - Cefdinir is a new, extended-spectrum, orally active, third-generation cephalosporin that is resistant to bacterial beta-lactamase production. To evaluate efficacy and safety of the antibiotic in maxillary sinusitis, its use was compared with amoxicillin/clavulanate (amox/clav), which is a well-accepted beta-lactamase-resistant antibiotic. In this investigator-blinded multicenter phase III clinical study, 569 patients were randomly assigned to one of three treatment regimens: one daily dose of cefdinir 600 mg (OD), cefdinir 300 mg every 12 h (BD), and amox/clav 500/125 mg every 8 h. All antibiotics were administered orally for 10 days. Maxillary sinusitis was documented by typical clinical signs and symptoms and was confirmed by X-ray imaging. Before treatment, the genus and species of any pathogens were determined from sinus aspirates. Cultures were tested for beta-lactmase production and in vitro resistance to cefdinir and amox/clav. The effectiveness of antibiotic treatment was evaluated 7-14 days after therapy and whether or not recurrent clinical symptoms or persistent infection was determined 21-35 days post-therapy. The appearance of any adverse events was classified as associated or not associated with the medication of the study. Present findings showed that the in vitro susceptibility of pathogens to cefdinir and amox/clav was similar. Cefdinir OD or BD was therapeutically as effective as or better than amox/clav, although cefdinir BD was not as useful as amox/clav clinically. Cefdinir OD and BD and amox/clav were well tolerated. The statistical incidence of adverse events was the same among the three treatment groups, although cefdinir OD treatment had significantly fewer treatment discontinuations due to adverse events than BD and amox/clav. PMID- 10839488 TI - Analysis of aromatic DNA adducts in laryngeal biopsies. AB - Epidemiological studies have confirmed the correlation between tobacco smoking, environmental pollution and the incidence of cancers of the respiratory tract. The occurrence of laryngeal cancer in Poland is relatively high compared to other European countries. Since 1969 the mortality related to larynx cancer appears to be increasing. Tobacco smoke contains an abundance of such carcinogenic compounds as polycyclic aromatic hydrocarbons (PAH), aromatic amines and N-nitrosoamines, which can react with DNA and form adducts. We analyzed aromatic DNA adducts in laryngeal tissues from patients with primary laryngeal, which was confirmed histopathologically to be squamous cell carcinoma. The group consisted of 33 patients (5 women and 28 men). Total laryngectomy was performed in patients. A detergentphenol method was used for DNA isolation. Aromatic DNA adducts were analyzed by a 32P-postlabelling technique with butanol extraction and high performance liquid chromatography. The presence of aromatic DNA adducts was demonstrated in all tissues. Large interindividual differences of DNA adduct levels were seen in each tissue studied. There was a higher mean level of DNA adducts in interarytenoid area non-tumors (51.96/10(8) +/- 91.71 NN) than in non tumor tissue elsewhere (46.91/10(8) +/- 46.36 NN) and tumor tissue (43.52/10(8) +/- 45.88 NN). Adduct levels were correlated with age, sex, cigarette smoking and TNM stage. PMID- 10839489 TI - Basaloid squamous cell carcinoma of the supraglottic larynx. AB - Basaloid squamous cell carcinoma (BSC) is regarded as a variant of squamous cell carcinoma, but displays distinct morphological and biological features as well as a different clinical course. The tumor is frequently seen in the head and neck and is preferentially located in the larynx, especially in supraglottic sites. Ten patients with BSC of the supraglottic larynx were treated from 1991 to 1995 at the Medical Faculty of the University of Istanbul. Results of treatment were compared retrospectively with a control group consisting of 44 patients with well differentiated squamous cell carcinomas. Ages, localizations, stages and treatment procedures were similar. In both groups mean survival, nodal involvement and distant metastases were comparable although the local (laryngeal) recurrence rate in patients with early supraglottic (T2) disease in the BSC group after conservative partial surgery was distinct compared to the control group (P < 0.05). These results indicate that conservative surgery should be assessed with caution in patients with BSC, and postoperative irradiation be taken into consideration. PMID- 10839491 TI - Sialolithiasis in children as a diagnostic dilemma. AB - Sialolithiasis is an uncommon disorder in childhood. Initially asymptomatic, symptoms may appear gradually. These can vary from moderate discomfort to severe pain with large glandular swelling accompanied by trismus. The correct interpretation of symptoms and a proper investigation for localization of salivary stones are important for effective treatment. A case of submandibular sialolithiasis in a 9-year-old girl is used to exemplify the problems had in clinical diagnosis. PMID- 10839490 TI - A potential role for arginase in recurrent infections and hypertrophy of tonsillar and adenoidal tissue. AB - Since arginase has been found to be an arginine-depleting and nitric oxide synthase-regulating enzyme, the present study was devised to examine hypertrophied and infected tonsil and adenoid arginase activity in relation to a metabolic arginase-nitric oxide pathway and its association with disease processes. Tissues were taken from 32 children undergoing adenotonsillectomy. There was a statistically significant difference between the two tissue enzyme activities, with tonsillar arginase activity being higher than the corresponding adenoidal tissue (P < 0.005). This suggests a potential role for tissue arginase activity as an outcome module and a contributing factor in chronic recurrent infection and hypertrophy of tonsillar and adenoidal tissues. PMID- 10839492 TI - A comparison of radiotherapy or radiochemotherapy with symptomatic treatment alone in patients with advanced head and neck carcinomas. AB - The choice of palliative treatment and the prognostic factors in unresectable head and neck cancer cases continue to be controversial. In the present study we compared the survival rates of untreated stage IV head and neck cancer patients with cases managed prospectively at A.C. Camargo Hospital for Cancer with neoadjuvant chemotherapy, concomitant chemotherapy or radiotherapy alone. Previous results had shown that while the type of treatment did not influence survival rates (P = 0.706), tumor response to treatment (whether complete, partial or none) significantly influenced survival (P = 0.00002). In the present study we compared the survival rates in the groups with untreated patients (who remained untreated until death) with the same demographic and clinical characteristics of patients receiving treatment. We found that there was a significant difference between the survival rates of the untreated group and those of the treated groups that was independent of the type of treatment performed (P < 0.00001) or the tumor response to treatment (P < 0.0001). PMID- 10839493 TI - The pathology and management of subglottic cancer. AB - Because there is still considerable controversy concerning the anatomical boundaries separating the three regions of the larynx, cancer of the subglottis remains difficult to manage. We have reviewed the numerous differences in the anatomical definitions used in the literature and the consequent differences in reported findings on the incidence of subglottic cancer and its classification. We have also summarized the pathology of subglottic malignant neoplastic lesions, their presenting symptoms and tendency for spread, and the use of imaging methods in its diagnosis. Suitable forms of treatment are discussed, as are considerations on prognosis. PMID- 10839494 TI - Metabolism of styrene by human liver and lung. AB - In mice, styrene is pneumotoxic, and there is some evidence of tumorigenicity. This toxicity is thought to be related to its bioactivation to styrene oxide in lung. To determine if human tissues have this capacity, the metabolism of styrene to styrene oxide was measured in human liver and lung microsomal preparations. Hepatic microsomes metabolized styrene to styrene oxide, but lung microsomes had essentially no activity. However, microsomes from both tissues metabolized benzene to phenol. The data suggest that human lung has low styrene metabolizing activity and may be much less of a target organ than in mouse. PMID- 10839495 TI - Behavior of free-ranging and captive American kestrels under electromagnetic fields. AB - Wild birds, particularly raptors, commonly use electrical transmission structures for nesting, perching, hunting, and roosting. Consequently, birds are exposed to electromagnetic fields (EMFs). The amount of time that wild reproducing American kestrels (Falco sparverius) were exposed to EMFs was determined, and the effects of EMFs on the behavior of captive reproducing kestrels were examined. Wild kestrels were exposed to EMFs for 25% to 75% of the observed time. On a 24-h basis, estimated EMF exposure of wild kestrels ranged from 71% during courtship, to 90% during incubation, similar to that experienced by captive kestrels (88% of a 24-h period). Additionally, captive kestrels were exposed to EMF levels experienced by wild kestrels nesting under 735-kV power lines. Captive EMF females were more active, more alert, and perched on the pen roof more frequently than control females during courtship. EMF females preened and rested less often during brood rearing. EMF male kestrels were more active than control males during courtship, and more alert during incubation. Increased activity of kestrels during courtship may be linked to changes in corticosterone, but likely not melatonin. Observed behavioral changes were unlikely to directly result in the better growth of nestlings and fledging success, or poorer hatching success, of the EMF group, as previously reported. Behavioral changes of captive EMF kestrels may be observed in wild kestrels. PMID- 10839496 TI - Methodology for characterizing distributions of incremental body burdens of 2,3,7,8-TCDD and DDE from breast milk in North American nursing infants. AB - A clear picture of ranges of doses of breast-milk contaminants experienced by nursing infants in North America has not yet been described, resulting in a significant gap in our understanding of potential health risks to infants from those contaminants. While point estimates of incremental dose have appeared in the published literature, these do not account for the wide variability in exposures experienced by nursing infants. This research expands on the current state of understanding of breast-milk contaminant exposure by characterizing distributions, rather than point estimates, of dose. Distributions of milk intake by nursing infants were characterized to examine intake of 2,3,7,8 tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD) and dichlorodiphenyl dichloroethane (DDE). The results indicate that, despite the uncertainties inherent in modeling incremental body burdens of chemicals from nursing, estimating incremental infant body burdens of lipophilic chemicals from breastfeeding using point estimates may result in overly conservative estimates of the contribution of breastfeeding to long-term body burdens of those chemicals in children. To develop reliable estimates of incremental body burden from nursing, depuration via lactation and half-life in the infant should be considered. Further, incremental infant body burdens of lipophilic chemicals increase rapidly at the start of lactation, but decrease after approximately 5 to 6 mo; by 2 yr postpartum, incremental body burdens have decreased substantially. Given the benefits afforded to infants who breastfeed, and because breastfeeding does not necessarily lead to significantly increased long-term body burdens in infants, breastfeeding should be encouraged and promoted. PMID- 10839497 TI - Effects of trimetazidine on oxidant/antioxidant status in trinitrobenzenesulfonic acid-induced chronic colitis. AB - Trimetazidine (TMZ), an anti-ischemic agent with proposed antioxidant properties, was used in a chronic colitis model in order to evaluate its effectiveness as a therapeutic agent in chronic colitis. Treatment of male Swiss Albino rats with ethanol (50%) and trinitrobenzenesulfonic acid (TNBS) (30 mg/kg) produced colitis as evidenced by histopathologic damage and inflammatory alterations, lipid peroxidation [increased malondialdehyde (MDA) levels], and enhanced neutrophil infiltration [increased myeloperoxidase (MPO) activity] without marked change in glutathione status. Administration of TMZ (5 mg/kg) to TNBS-treated rats failed to affect the TNBS-induced changes in histopathology and MPO activities. Unexpectedly, intrarectal (i.r.) administration of TMZ significantly elevated colonic MDA levels to a greater extent than TNBS alone. Intraperitoneal (i.p.) TMZ treatment seemed to increase total glutathione (tGSH), GSH, and GSH/GSSG values. In conclusion, our results demonstrated that (a) i.r. administration of ethanol and TNBS is an effective way of inducing a chronic colitis model, (b) inflammation and lipid peroxidation augment tissue damage in the chronic colitis model, (c) i.p. TMZ treatment significantly inhibits MDA production in the chronic colitis model, (d) TMZ treatment is more effective via the i.p. compared to i.r. route, and (e) TMZ seems to show its antioxidant effect via preserving the tissue's GSH/GSSG ratios. PMID- 10839499 TI - Powerful visions for astronomy PMID- 10839498 TI - Determination of biokinetic interactions in chemical mixtures using real-time breath analysis and physiologically based pharmacokinetic modeling. AB - Regulatory agencies are challenged to conduct risk assessments on chemical mixtures without full information on toxicological interactions that may occur at real-world, low-dose exposure levels. The present study was undertaken to investigate the pharmacokinetic impact of low-dose coexposures to toluene and trichloroethylene in vivo in male F344 rats using a real-time breath analysis system coupled with physiologically based pharmacokinetic (PBPK) modeling. Rats were exposed to compounds alone or as a binary mixture, at low (5 to 25 mg/kg) or high (240 to 800 mg/kg) dose levels. Exhaled breath from the exposed animals was monitored for the parent compounds and a PBPK model was used to analyze the data. At low doses, exhaled breath kinetics from the binary mixture exposure compared with those obtained during single exposures, thus indicating that no metabolic interaction occurred with these low doses. In contract, at higher doses the binary PBPK model simulating independent metabolism was found to underpredict the exhaled breath concentration, suggesting an inhibition of metabolism. Therefore the binary mixture PBPK model was used to compare the measured exhaled breath levels from high- and low-dose exposures with the predicted levels under various metabolic interaction simulations (competitive, noncompetitive, or uncompetitive inhibition). Of these simulations, the optimized competitive metabolic interaction description yielded a Ki value closest to the Km of the inhibitor solvent, indicating that competitive inhibition is the most plausible type of metabolic interaction between these two solvents. PMID- 10839500 TI - Values of the abstract PMID- 10839502 TI - France sets up elite Internet school PMID- 10839501 TI - US astronomers draw up their wish list for a decade of funding PMID- 10839503 TI - Britain asks to join European Southern Observatory PMID- 10839504 TI - Mathematicians chase the seven million-dollar proofs PMID- 10839505 TI - BioMed Central boosted by editorial board. PMID- 10839506 TI - Russian PM supports science PMID- 10839507 TI - Congress wakes up to climate change PMID- 10839508 TI - French unions upset by plans for physiology centre. PMID- 10839510 TI - Tax blow leaves Australian science reeling PMID- 10839509 TI - Bacterial AIDS vaccine ready for testing. PMID- 10839511 TI - Sanger Centre. Changing of the guard. PMID- 10839512 TI - EarthScope. Deep thoughts PMID- 10839513 TI - It's time to work together and stop duplicating conservation efforts... PMID- 10839514 TI - Following Africa's lead in setting priorities. PMID- 10839516 TI - Many paths to enlightenment PMID- 10839515 TI - Community groups could show Unesco the way PMID- 10839517 TI - Brochure. The apocalypse: a great day out for the whole family PMID- 10839518 TI - Loophole for snowball Earth. PMID- 10839519 TI - Cancer. New link in a web of human genes. PMID- 10839520 TI - Star formation. Three's a crowd PMID- 10839522 TI - Seismology. Inner core takes another turn PMID- 10839523 TI - Carbon fixation. PMID- 10839521 TI - Genomic imprinting. Silence across the border. PMID- 10839524 TI - Applied mathematics. The power of design PMID- 10839525 TI - Self-regulating synapses. PMID- 10839526 TI - DNA repair. The bases for Cockayne syndrome. PMID- 10839527 TI - Wireless capsule endoscopy. PMID- 10839528 TI - Non-thermal heat-shock response to microwaves. PMID- 10839529 TI - Structural biology. Proton-powered turbine of a plant motor. PMID- 10839530 TI - Roles of PPARs in health and disease. AB - In developed societies, chronic diseases such as diabetes, obesity, atherosclerosis and cancer are responsible for most deaths. These ailments have complex causes involving genetic, environmental and nutritional factors. There is evidence that a group of closely related nuclear receptors, called peroxisome proliferator-activated receptors (PPARs), may be involved in these diseases. This, together with the fact that PPAR activity can be modulated by drugs such as thiazolidinediones and fibrates, has instigated a huge research effort into PPARs. Here we present the latest developments in the PPAR field, with particular emphasis on the physiological function of PPARs during various nutritional states, and the possible role of PPARs in several chronic diseases. PMID- 10839531 TI - Neoproterozoic 'snowball Earth' simulations with a coupled climate/ice-sheet model. AB - Ice sheets may have reached the Equator in the late Proterozoic era (600-800 Myr ago), according to geological and palaeomagnetic studies, possibly resulting in a 'snowball Earth'. But this period was a critical time in the evolution of multicellular animals, posing the question of how early life survived under such environmental stress. Here we present computer simulations of this unusual climate stage with a coupled climate/ice-sheet model. To simulate a snowball Earth, we use only a reduction in the solar constant compared to present-day conditions and we keep atmospheric CO2 concentrations near present levels. We find rapid transitions into and out of full glaciation that are consistent with the geological evidence. When we combine these results with a general circulation model, some of the simulations result in an equatorial belt of open water that may have provided a refugium for multicellular animals. PMID- 10839532 TI - A triplet of differently shaped spin-zero states in the atomic nucleus 186Pb AB - Understanding the fundamental excitations of many-fermion systems is of significant current interest. In atomic nuclei with even numbers of neutrons and protons, the low-lying excitation spectrum is generally formed by nucleon pair breaking and nuclear vibrations or rotations. However, for certain numbers of protons and neutrons, a subtle rearrangement of only a few nucleons among the orbitals at the Fermi surface can result in a different elementary mode: a macroscopic shape change. The first experimental evidence for this phenomenon came from the observation of shape coexistence in 16O (ref. 4). Other unexpected examples came with the discovery of fission isomers and super-deformed nuclei. Here we find experimentally that the lowest three states in the energy spectrum of the neutron deficient nucleus 186Pb are spherical, oblate and prolate. The states are populated by the alpha-decay of a parent nucleus; to identify them, we combine knowledge of the particular features of this decay with sensitive measurement techniques (a highly efficient velocity filters with strong background reduction, and an extremely selective recoil-alpha-electron coincidence tagging methods). The existence of this apparently unique shape triplet is permitted only by the specific conditions that are met around this particular nucleus. PMID- 10839533 TI - Microdomain patterns from directional eutectic solidification and epitaxy AB - Creating a regular surface pattern on the nanometre scale is important for many technological applications, such as the periodic arrays constructed by optical microlithography that are used as separation media in electrophoresis, and island structures used for high-density magnetic recording devices. Block copolymer patterns can also be used for lithography on length scales below 30 nanometres (refs 3-5). But for such polymers to prove useful for thin-film technologies, chemically patterned surfaces need to be made substantially defect-free over large areas, and with tailored domain orientation and periodicity. So far, control over domain orientation has been achieved by several routes, using electric fields, temperature gradients, patterned substrates and neutral confining surfaces. Here we describe an extremely fast process that leads the formation of two-dimensional periodic thin films having large area and uniform thickness, and which possess vertically aligned cylindrical domains each containing precisely one crystalline lamella. The process involves rapid solidification of a semicrystalline block copolymer from a crystallizable solvent between glass substrates using directional solidification and epitaxy. The film is both chemically and structurally periodic, thereby providing new opportunities for more selective and versatile nanopatterned surfaces. PMID- 10839534 TI - Large-scale synthesis of a silicon photonic crystal with a complete three dimensional bandgap near 1.5 micrometres AB - Photonic technology, using light instead of electrons as the information carrier, is increasingly replacing electronics in communication and information management systems. Microscopic light manipulation, for this purpose, is achievable through photonic bandgap materials, a special class of photonic crystals in which three dimensional, periodic dielectric constant variations controllably prohibit electromagnetic propagation throughout a specified frequency band. This can result in the localization of photons, thus providing a mechanism for controlling and inhibiting spontaneous light emission that can be exploited for photonic device fabrication. In fact, carefully engineered line defects could act as waveguides connecting photonic devices in all-optical microchips, and infiltration of the photonic material with suitable liquid crystals might produce photonic bandgap structures (and hence light-flow patterns) fully tunable by an externally applied voltage. However, the realization of this technology requires a strategy for the efficient synthesis of high-quality, large-scale photonic crystals with photonic bandgaps at micrometre and sub-micrometre wavelengths, and with rationally designed line and point defects for optical circuitry. Here we describe single crystals of silicon inverse opal with a complete three dimensional photonic bandgap centred on 1.46 microm, produced by growing silicon inside the voids of an opal template of dose-packed silica spheres that are connected by small 'necks' formed during sintering, followed by removal of the silica template. The synthesis method is simple and inexpensive, yielding photonic crystals of pure silicon that are easily integrated with existing silicon-based microelectronics. PMID- 10839535 TI - Deep convective clouds with sustained supercooled liquid water down to -37.5 degrees C AB - In cirrus and orographic wave clouds, highly supercooled water has been observed in small quantities (less than 0.15 g m(-3)). This high degree of supercooling was attributed to the small droplet size and the lack of ice nuclei at the heights of these clouds. For deep convective clouds, which have much larger droplets near their tops and which take in aerosols from near the ground, no such measurements have hitherto been reported. However, satellite data suggest that highly supercooled water (down to -38 degrees C) frequently occurs in vigorous continental convective storms. Here we report in situ measurements in deep convective clouds from an aircraft, showing that most of the condensed water remains liquid down to -37.5 degrees C. The droplets reach a median volume diameter of 17 microm and amount to 1.8 gm(-3), one order of magnitude more than previously reported. At slightly colder temperatures only ice was found, suggesting homogeneous freezing. Because of the poor knowledge of mixed-phase cloud processes, the simulation of clouds using numerical models is difficult at present. Our observations will help to understand these cloud processes, such as rainfall, hail, and cloud electrification, together with their implications for the climate system. PMID- 10839536 TI - Past temperature and delta18O of surface ocean waters inferred from foraminiferal Mg/Ca ratios. AB - Determining the past record of temperature and salinity of ocean surface waters is essential for understanding past changes in climate, such as those which occur across glacial-interglacial transitions. As a useful proxy, the oxygen isotope composition (delta18O) of calcite from planktonic foraminifera has been shown to reflect both surface temperature and seawater delta18O, itself an indicator of global ice volume and salinity. In addition, magnesium/calcium (Mg/Ca) ratios in foraminiferal calcite show a temperature dependence due to the partitioning of Mg during calcification. Here we demonstrate, in a field-based calibration experiment, that the variation of Mg/Ca ratios with temperature is similar for eight species of planktonic foraminifera (when accounting for Mg dissolution effects). Using a multi-species record from the Last Glacial Maximum in the North Atlantic Ocean we found that past temperatures reconstructed from Mg/Ca ratios followed the two other palaeotemperature proxies: faunal abundance and alkenone saturation. Moreover, combining Mg/Ca and delta18O data from the same faunal assemblage, we show that reconstructed surface water delta18O from all foraminiferal species record the same glacial-interglacial change--representing changing hydrography and global ice volume. This reinforces the potential of this combined technique in probing past ocean-climate interactions. PMID- 10839537 TI - Slow differential rotation of the Earth's inner core indicated by temporal changes in scattering AB - The finding that the Earth's inner core might be rotating faster than the mantle has important implications for our understanding of core processes, including the generation of the Earth's magnetic field. But the reported signal is subtle--a change of about 0.01 s per year in the separation of two seismic waves with differing paths through the core. Subsequent studies of such data have generally supported the conclusion that differential rotation exists, but the difficulty of accurately locating historic earthquakes and possible biases induced by strong lateral variations in structure near the core-mantle boundary have raised doubt regarding the proposed inner-core motion. Also, a study of free oscillations constrained the motion to be relatively small compared to previous estimates and it has been proposed that the interaction of inner-core boundary topography and mantle heterogeneity might lock the inner core to the mantle. The recent detection of seismic waves scattered in the inner core suggests a simple test of inner-core motion. Here we compare scattered waves recorded in Montana, USA, from two closely located nuclear tests at Novaya Zemlya, USSR, in 1971 and 1974. The data show small but coherent changes in scattering which point toward an inner core differential rotation rate of 0.15 degrees per year--consistent with constraints imposed by the free-oscillation data. PMID- 10839538 TI - Invasion sequence affects predator-prey dynamics in a multi-species interaction. AB - Ecologists seek to understand the rules that govern the assembly, coexistence and persistence of communities of interacting species. There is, however, a variety of sequences in which a multi-species community can be assembled--unlike more familiar one- and two-species systems. Ecological systems can exhibit contrasting dynamics depending on initial conditions, but studies have been focused on simple communities initiated at different densities, not on multi-species communities constructed in different sequences. Investigations of permanence and convergence in ecological communities have been concerned with the flux of whole species (presence or absence) but have not addressed the central issues concerning the dynamics exhibited by individual species in particular interactions. Here we examine data for replicated three-species systems and demonstrate that the dynamic trajectories of both a predator and its prey within the system are determined by the sequence in which it is constructed, and that for one construction-sequence alternative dynamic patterns are possible. PMID- 10839539 TI - Chromosomal evolution in Saccharomyces. AB - The chromosomal speciation model invokes chromosomal rearrangements as the primary cause of reproductive isolation. In a heterozygous carrier, chromosomes bearing reciprocal translocations mis-segregate at meiosis, resulting in reduced fertility or complete sterility. Thus, chromosomal rearrangements act as a post zygotic isolating mechanism. Reproductive isolation in yeast is due to post zygotic barriers, as many species mate successfully but the hybrids are sterile. Reciprocal translocations are thought to be the main form of large-scale rearrangement since the hypothesized duplication of the whole yeast genome 10(8) years ago. To test the chromosomal speciation model in yeast, we have characterized chromosomal translocations among the genomes of six closely related species in the Saccharomyces 'sensu stricto' complex. Here we show that rearrangements have occurred between closely related species, whereas more distant ones have colinear genomes. Thus, chromosomal rearrangements are not a prerequisite for speciation in yeast and the rate of formation of translocations is not constant. These rearrangements appear to result from ectopic recombination between Ty elements or other repeated sequences. PMID- 10839540 TI - Synaptic activity at calcium-permeable AMPA receptors induces a switch in receptor subtype. AB - Activity-dependent change in the efficacy of transmission is a basic feature of many excitatory synapses in the central nervous system. The best understood postsynaptic modification involves a change in responsiveness of AMPAR (alpha amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptor)-mediated currents following activation of NMDA (N-methyl-D-aspartate) receptors or Ca2+-permeable AMPARs. This process is thought to involve alteration in the number and phosphorylation state of postsynaptic AMPARs. Here we describe a new form of synaptic plasticity--a rapid and lasting change in the subunit composition and Ca2+ permeability of AMPARs at cerebellar stellate cell synapses following synaptic activity. AMPARs lacking the edited GluR2 subunit not only exhibit high Ca2+ permeability but also are blocked by intracellular polyamines. These properties have allowed us to follow directly the involvement of GluR2 subunits in synaptic transmission. Repetitive synaptic activation of Ca2+-permeable AMPARs causes a rapid reduction in Ca2+ permeability and a change in the amplitude of excitatory postsynaptic currents, owing to the incorporation of GluR2-containing AMPARs. Our experiments show that activity-induced Ca2+ influx through GluR2 lacking AMPARs controls the targeting of GluR2-containing AMPARs, implying the presence of a self-regulating mechanism. PMID- 10839541 TI - Vagus nerve stimulation attenuates the systemic inflammatory response to endotoxin. AB - Vertebrates achieve internal homeostasis during infection or injury by balancing the activities of proinflammatory and anti-inflammatory pathways. Endotoxin (lipopolysaccharide), produced by all gram-negative bacteria, activates macrophages to release cytokines that are potentially lethal. The central nervous system regulates systemic inflammatory responses to endotoxin through humoral mechanisms. Activation of afferent vagus nerve fibres by endotoxin or cytokines stimulates hypothalamic-pituitary-adrenal anti-inflammatory responses. However, comparatively little is known about the role of efferent vagus nerve signalling in modulating inflammation. Here, we describe a previously unrecognized, parasympathetic anti-inflammatory pathway by which the brain modulates systemic inflammatory responses to endotoxin. Acetylcholine, the principle vagal neurotransmitter, significantly attenuated the release of cytokines (tumour necrosis factor (TNF), interleukin (IL)-1beta, IL-6 and IL-18), but not the anti inflammatory cytokine IL-10, in lipopolysaccharide-stimulated human macrophage cultures. Direct electrical stimulation of the peripheral vagus nerve in vivo during lethal endotoxaemia in rats inhibited TNF synthesis in liver, attenuated peak serum TNF amounts, and prevented the development of shock. PMID- 10839542 TI - Targeted destabilization of HY5 during light-regulated development of Arabidopsis. AB - Arabidopsis seedlings display contrasting developmental patterns depending on the ambient light. Seedlings grown in the light develop photomorphogenically, characterized by short hypocotyls and expanded green cotyledons. In contrast, seedlings grown in darkness become etiolated, with elongated hypocotyls and dosed cotyledons on an apical hook. Light signals, perceived by multiple photoreceptors and transduced to downstream regulators, dictate the extent of photomorphogenic development in a quantitative manner. Two key downstream components, COP1 and HY5, act antagonistically in regulating seedling development. HY5 is a bZIP transcription factor that binds directly to the promoters of light-inducible genes, promoting their expression and photomorphogenic development. COP1 is a RING-finger protein with WD-40 repeats whose nuclear abundance is negatively regulated by light. COP1 interacts directly with HY5 in the nucleus to regulate its activity negatively. Here we show that the abundance of HY5 is directly correlated with the extent of photomorphogenic development, and that the COP1-HY5 interaction may specifically target HY5 for proteasome-mediated degradation in the nucleus. PMID- 10839543 TI - Genomic rearrangement in NEMO impairs NF-kappaB activation and is a cause of incontinentia pigmenti. The International Incontinentia Pigmenti (IP) Consortium. AB - Familial incontinentia pigmenti (IP; MIM 308310) is a genodermatosis that segregates as an X-linked dominant disorder and is usually lethal prenatally in males. In affected females it causes highly variable abnormalities of the skin, hair, nails, teeth, eyes and central nervous system. The prominent skin signs occur in four classic cutaneous stages: perinatal inflammatory vesicles, verrucous patches, a distinctive pattern of hyperpigmentation and dermal scarring. Cells expressing the mutated X chromosome are eliminated selectively around the time of birth, so females with IP exhibit extremely skewed X inactivation. The reasons for cell death in females and in utero lethality in males are unknown. The locus for IP has been linked genetically to the factor VIII gene in Xq28 (ref. 3). The gene for NEMO (NF-kappaB essential modulator)/IKKgamma (IkappaB kinase-gamma) has been mapped to a position 200 kilobases proximal to the factor VIII locus. NEMO is required for the activation of the transcription factor NF-kappaB and is therefore central to many immune, inflammatory and apoptotic pathways. Here we show that most cases of IP are due to mutations of this locus and that a new genomic rearrangement accounts for 80% of new mutations. As a consequence, NF-kappaB activation is defective in IP cells. PMID- 10839544 TI - Functional link between ataxia-telangiectasia and Nijmegen breakage syndrome gene products. AB - Ataxia-telangiectasia (A-T) and Nijmegen breakage syndrome (NBS) are recessive genetic disorders with susceptibility to cancer and similar cellular phenotypes. The protein product of the gene responsible for A-T, designated ATM, is a member of a family of kinases characterized by a carboxy-terminal phosphatidylinositol 3 kinase-like domain. The NBS1 protein is specifically mutated in patients with Nijmegen breakage syndrome and forms a complex with the DNA repair proteins Rad50 and Mrel1. Here we show that phosphorylation of NBS1, induced by ionizing radiation, requires catalytically active ATM. Complexes containing ATM and NBS1 exist in vivo in both untreated cells and cells treated with ionizing radiation. We have identified two residues of NBS1, Ser 278 and Ser 343 that are phosphorylated in vitro by ATM and whose modification in vivo is essential for the cellular response to DNA damage. This response includes S-phase checkpoint activation, formation of the NBS1/Mrel1/Rad50 nuclear foci and rescue of hypersensitivity to ionizing radiation. Together, these results demonstrate a biochemical link between cell-cycle checkpoints activated by DNA damage and DNA repair in two genetic diseases with overlapping phenotypes. PMID- 10839545 TI - ATM phosphorylation of Nijmegen breakage syndrome protein is required in a DNA damage response. AB - Nijmegen breakage syndrome (NBS) is characterized by extreme radiation sensitivity, chromosomal instability and cancer. The phenotypes are similar to those of ataxia telangiectasia mutated (ATM) disease, where there is a deficiency in a protein kinase that is activated by DNA damage, indicating that the Nbs and Atm proteins may participate in common pathways. Here we report that Nbs is specifically phosphorylated in response to gamma-radiation, ultraviolet light and exposure to hydroxyurea. Phosphorylation of Nbs mediated by gamma-radiation, but not that induced by hydroxyurea or ultraviolet light, was markedly reduced in ATM cells. In vivo, Nbs was phosphorylated on many serine residues, of which S343, S397 and S615 were phosphorylated by Atm in vitro. At least two of these sites were underphosphorylated in ATM cells. Inactivation of these serines by mutation partially abrogated Atm-dependent phosphorylation. Reconstituting NBS cells with a mutant form of Nbs that cannot be phosphorylated at selected, ATM-dependent serine residues led to a specific reduction in clonogenic survival after gamma radiation. Thus, phosphorylation of Nbs by Atm is critical for certain responses of human cells to DNA damage. PMID- 10839546 TI - Methylation of a CTCF-dependent boundary controls imprinted expression of the Igf2 gene. AB - The expression of the insulin-like growth factor 2 (Igf2) and H19 genes is imprinted. Although these neighbouring genes share an enhancer, H19 is expressed only from the maternal allele, and Igf2 only from the paternally inherited allele. A region of paternal-specific methylation upstream of H19 appears to be the site of an epigenetic mark that is required for the imprinting of these genes. A deletion within this region results in loss of imprinting of both H19 and Igf2 (ref. 5). Here we show that this methylated region contains an element that blocks enhancer activity. The activity of this element is dependent upon the vertebrate enhancer-blocking protein CTCF. Methylation of CpGs within the CTCF binding sites eliminates binding of CTCF in vitro, and deletion of these sites results in loss of enhancer-blocking activity in vivo, thereby allowing gene expression. This CTCF-dependent enhancer-blocking element acts as an insulator. We suggest that it controls imprinting of Igf2. The activity of this insulator is restricted to the maternal allele by specific DNA methylation of the paternal allele. Our results reveal that DNA methylation can control gene expression by modulating enhancer access to the gene promoter through regulation of an enhancer boundary. PMID- 10839547 TI - CTCF mediates methylation-sensitive enhancer-blocking activity at the H19/Igf2 locus. AB - The Insulin-like growth factor 2 (Igf2) and H19 genes are imprinted, resulting in silencing of the maternal and paternal alleles, respectively. This event is dependent upon an imprinted-control region two kilobases upstream of H19 (refs 1, 2). On the paternal chromosome this element is methylated and required for the silencing of H19 (refs 2-4). On the maternal chromosome the region is unmethylated and required for silencing of the Igf2 gene 90 kilobases upstream. We have proposed that the unmethylated imprinted-control region acts as a chromatin boundary that blocks the interaction of Igf2 with enhancers that lie 3' of H19 (refs 5, 6). This enhancer-blocking activity would then be lost when the region was methylated, thereby allowing expression of Igf2 paternally. Here we show, using transgenic mice and tissue culture, that the unmethylated imprinted control regions from mouse and human H19 exhibit enhancer-blocking activity. Furthermore, we show that CTCF, a zinc finger protein implicated in vertebrate boundary function, binds to several sites in the unmethylated imprinted-control region that are essential for enhancer blocking. Consistent with our model, CTCF binding is abolished by DNA methylation. This is the first example, to our knowledge, of a regulated chromatin boundary in vertebrates. PMID- 10839548 TI - Lowering the age for influenza vaccination. PMID- 10839549 TI - Genetic testing. PMID- 10839550 TI - Combination antihypertensive drug therapy: a therapeutic option long overdue. PMID- 10839551 TI - Child abuse: the physician's role in alleviating a growing problem. PMID- 10839552 TI - Treatment of prostatitis. AB - The term prostatitis is applied to a series of disorders, ranging from acute bacterial infection to chronic pain syndromes, in which the prostate gland is inflamed. Patients present with a variety of symptoms, including urinary obstruction, fever, myalgias, decreased libido or impotence, painful ejaculation and low-back and perineal pain. Physical examination often fails to clarify the cause of the pain. Cultures and microscopic examination of urine and prostatic secretions before and after prostatic massage may help differentiate prostatitis caused by infection from prostatitis with other causes. Because the rate of occult infection is high, a therapeutic trial of antibiotics is often in order even when patients do not appear to have bacterial prostatitis. If the patient responds to therapy, antibiotics are continued for at least three to four weeks, although some men require treatment for several months. A patient who does not respond might be evaluated for chronic nonbacterial prostatitis, in which nonsteroidal anti-inflammatory drugs, alpha-blocking agents, anticholinergic agents or other therapies may provide symptomatic relief. PMID- 10839553 TI - Age-related macular degeneration: update for primary care. AB - Age-related macular degeneration is the leading cause of severe vision loss among the elderly. In this condition, central vision is lost, but peripheral vision almost always remains intact. Affected persons rarely require canes or guide dogs. The diagnosis of age-related macular degeneration is based on symptoms and ophthalmoscopic findings, and the disease can be classified into atrophic and exudative forms. The two currently proven treatments are laser photocoagulation and photodynamic therapy, but these measures are effective in only a small fraction of eyes with the exudative form of macular degeneration. Vision rehabilitation can help patients maximize their remaining vision and adapt so that they can perform activities of daily living. Families need encouragement in providing support and helping patients adjust to being partially sighted. PMID- 10839554 TI - Combination antihypertensive drugs: recommendations for use. AB - The recommendation for first-line therapy for hypertension remains a beta blocker or diuretic given in a low dosage. A target blood pressure of less than 140/90 mm Hg is achieved in about 50 percent of patients treated with monotherapy; two or more agents from different pharmacologic classes are often needed to achieve adequate blood pressure control. Single-dose combination antihypertension therapy is an important option that combines efficacy of blood pressure reduction and a low side effect profile with convenient once-daily dosing to enhance compliance. Combination antihypertensives include combined agents from the following pharmacologic classes: diuretics and potassium-sparing diuretics, beta blockers and diuretics, angiotensin-converting enzyme (ACE) inhibitors and diuretics, angiotensin-II antagonists and diuretics, and calcium channel blockers and ACE inhibitors. PMID- 10839555 TI - Evaluation of physical abuse in children. AB - Family physicians who are involved in the care of children are likely to encounter child abuse and should be able to recognize its common presentations. A history that is inconsistent with the patient's injuries is the hallmark of physical abuse. A pattern of physical findings, including bruises and fractures in areas unlikely to be accidentally injured, patterned bruises from objects, and circumferential burns or bruises in children not yet mobile, should be viewed as suspicious for child abuse. Family physicians who suspect physical abuse are mandated to make a report to the state child protective services agency and to assure the ongoing safety of the child. PMID- 10839556 TI - Evaluation of pregnant women exposed to respiratory viruses. AB - Prenatal patients are often exposed to respiratory viruses at home and at work. Understandably, these patients may be concerned and want immediate answers and advice from their physicians. While most women who are exposed to chickenpox are immune, serologic testing can be performed and susceptible patients can be treated with varicella-zoster immune globulin. If the prenatal patient is infected with the varicella-zoster virus, the risk of fetal manifestations is less than 2 percent. Women who have been exposed to fifth disease can undergo serologic testing to determine the likelihood of infection. If the prenatal patient becomes infected with fifth disease during the first 20 weeks of gestation, the risk of fetal manifestations is about 9 percent and includes nonimmune hydrops and death. Cytomegalovirus, which is the most common congenital infection, is generally asymptomatic in the mother. Infected fetuses have a 25 percent chance of developing early or late neurologic manifestations. The evidence of harm from other common respiratory viruses is inconsistent. PMID- 10839557 TI - The painful shoulder: part I. Clinical evaluation. AB - Family physicians need to understand diagnostic and treatment strategies for common causes of shoulder pain. We review key elements of the history and physical examination and describe maneuvers that can be used to reach an appropriate diagnosis. Examination of the shoulder should include inspection, palpation, evaluation of range of motion and provocative testing. In addition, a thorough sensorimotor examination of the upper extremity should be performed, and the neck and elbow should be evaluated. PMID- 10839558 TI - Diagnosis and treatment of atrophic vaginitis. AB - Up to 40 percent of postmenopausal women have symptoms of atrophic vaginitis. Because the condition is attributable to estrogen deficiency, it may occur in premenopausal women who take antiestrogenic medications or who have medical or surgical conditions that result in decreased levels of estrogen. The thinned endometrium and increased vaginal pH level induced by estrogen deficiency predispose the vagina and urinary tract to infection and mechanical weakness. The earliest symptoms are decreased vaginal lubrication, followed by other vaginal and urinary symptoms that may be exacerbated by superimposed infection. Once other causes of symptoms have been eliminated, treatment usually depends on estrogen replacement. Estrogen replacement therapy may be provided systemically or locally, but the dosage and delivery method must be individualized. Vaginal moisturizers and lubricants, and participation in coitus may also be beneficial in the treatment of women with atrophic vaginitis. PMID- 10839559 TI - Clinical review of recent findings on the awareness, diagnosis and treatment of depression. PMID- 10839560 TI - Synergistic disaggregation of platelets by the products of endothelial cells or their analogs. AB - It is known that some products of endothelial cells or their analogs can attenuate the platelet aggregation response and initiate the platelet disaggregation response. Since platelets are involved in the initiation of many clinically important occlusive vascular diseases, we hypothesized that the endothelial cell products act synergistically to disperse platelet aggregates. In this study we examined the synergistic platelet disaggregating effects among the products of endothelial cells. We used urokinase, prostaglandin I2 (PGI2), and sodium nitroprusside (SNP) (which is the chemical substitute as nitric oxide(NO) donor) for endothelium-derived relaxing factor (EDRF). Platelet disaggregation rate was increased in a dose-dependent manner and decreased in a time-dependent manner, and the combined use of two or three agents had synergistic effects on platelet disaggregation. Furthermore, flow cytometric analysis showed decreases in the binding of fibrinogen to activated platelets by the addition of PGI2 or SNP. These data revealed that these products or their analogs could inactivate the activated platelets or aggregated platelets by detaching fibrinogen from platelets. In addition our data revealed that PGI2 and SNP can act synergistically with fibrinolytic agents. These findings suggest a potential strategy for improving the efficacy of thrombolytic therapy by a combination of these products or their substitutes. PMID- 10839561 TI - Effects of obstructive jaundice on the antioxidative capacity of human red blood cells. AB - Transient haemolysis and shortened erythrocyte lifespan are reported in association with extrahepatic biliary tract obstruction. An increase in lipid peroxidation has been noted as evidence of oxidative damage in red cells due to cholestasis. The influence of surgical relief on the antioxidative capacity of the erythrocyte is less well defined. The ability of erythrocytes to regenerate the antioxidative capacity after side-to-side choledo-choduodenostomy was assessed by measuring the two principal antioxidant enzymes, namely superoxide dismutase (SOD) and catalase (CAT), as well as the glutathione (GSH) content in the red blood cells (RBC) taken from patients with obstructive jaundice. A comparison of patients and healthy volunteers revealed a consistent decrease in enzyme activities (pSOD = 0.01, pCAT = 0.0002) and glutathione concentrations (PGSH = 0.0000) in cholestatic patients. Statistical analysis proved a clear correlation between the surgical relief of common bile duct obstruction and restored antioxidative capacity of red cells. These observations suggest that the red cells from patients with multiple common bile duct stones almost completely regenerated their antioxidative capacity four weeks after side-to-side choledochoduodenostomy. PMID- 10839562 TI - Effects of endothelial cells and mononuclear leukocytes on platelet aggregation. AB - Although normal vascular endothelium prevents adhesion and aggregation of platelets by the release of nitric oxide (NO) and prostacyclin, the endothelium exposed to oxidized low density lipoprotein (LDL) may be damaged, damage that may include a reduction in its secretory capacity. On the other hand, mononuclear leukocytes (ML) can also release NO to inhibit platelet aggregation. Platelet aggregation was measured with a Chrono-Log aggregometer (USA) in the presence of cultured endothelial cells (EC) or isolated ML, stimulated by collagen. Platelet aggregation was markedly inhibited (28.5 +/- 5.0 versus control 92.2 +/- 1.7%), when EC were added to platelets. Pre-incubation of the EC with 10 micromol/l of indomethacin or 300 micromol/l of L-NG-monomethyl-arginine (L-NMMA) for 6 h substantially reduced their anti-aggregating activity (72.8 +/- 5.1 and 76.6 +/- 7.7%, respectively). However, incubation of the EC in culture with 10 micromol/l of lysophosphatidyl choline (LPC) for 6 h did not affect the anti-aggregating capacity of EC (LPC 28.1 +/- 5.5 vs. EC 28.5 +/- 5.0%). Non-stimulated ML also inhibited (43.2 + 5.9 vs. control 69.2 3.2%) the platelet aggregation induced by collagen. The inhibition was dependent on the number of ML added. In addition, it was enhanced by increasing the concentration of collagen from 2.5 to 10 microg/ml. Platelet aggregation is inhibited mainly by the EC, however, blood ML also inhibit platelet aggregation. Incubation of the EC in culture with LPC did not affect the anti-aggregating capacity of the EC. PMID- 10839563 TI - Investigation of platelet aggregation by impedance and optic methods in children with iron deficiency anaemia. AB - Although it is known that platelet count is altered in iron deficiency anaemia (IDA), the qualitative extent of this interference is not well documented. In the present study we investigated platelet aggregation (PA) by impedance and optic methods in IDA. Forty-seven patients (plasma group: 16 boys, 9 girls and whole blood group: 11 boys, 11 girls) with IDA and thirty-one healthy children (plasma group: 6 boys, 10 girls and whole blood group: 6 boys, 9 girls) were enrolled into the study. Template bleeding times were measured by the Ivy method in all children. In the control group whole blood count, serum iron levels, bleeding time and PA were determined. After basal PA was determined in the patients and controls, ferrous sulphate was orally administered to the patients at a dose of 6 mg/kg/24 h for three months. Then, PA tests were performed again in the IDA (test group) patients. Ristocetin-induced PA was suppressed in both plasma and whole blood groups. Inhibition by both collagen (p < 0.05) and ristocetin (p < 0.001) induced PA was determined by the optic method. Similarly in PA measured by the impedance method a suppression to adenosine diphosphate (p < 0.001) and to ristocetin (p < 0.01) was found. However, no significant alteration was observed in the bleeding time. All defective responses were reversed by the iron supplementation therapy. In addition, a significant correlation was found between some parameters of PA and several haematological values. In conclusion, although defective PA responses cannot be clinically demonstrated in patients with IDA, this suppression of PA may be detected by laboratory examination. Therefore, it is advised that care should be taken when using anti-aggregant agents in IDA. PMID- 10839564 TI - The incidence of pseudothrombocytopenia in automatic blood analyzers. AB - The aim of the present work was to undertake an assessment of the incidence of pseudothrombocytopenia (PTCP) in patients referred for evaluation of thrombocytopenia in an outpatient hematology clinic. METHODS: Prospective assessment of 60 consecutive cases with platelet count < 100 x 10(9)/l in a hematology clinic during a 2-year period. RESULTS: PTCP was the second most common cause for low platelet count, with an incidence of 17%. Platelet count of patients with PTCP at presentation was 42 +/- 22 x 10(9)/l, and when re-analyzed on fresh samples, 208 +/- 39 x 10(9)/l. The relatively high prevalence of pseudothrombocytopenia in our series was due to a lack of microscopic inspection of the blood smear in the primary care laboratories and considerable delay in sample processing. CONCLUSIONS: PTCP should be considered in the assessment of low platelet count. While decreasing the transfer time of blood specimens may decrease PTCP incidence, microscopic inspection of the blood smear may avoid erroneous diagnosis of thrombocytopenia. PMID- 10839565 TI - Novel rosette formation of murine spleen cells with autologous red blood cells. AB - Spleen cells of normal BALB/c mice formed rosettes with autologous red blood cells, and the formation was calcium ion dependent. Peritoneal exudate cells, bone marrow cells and thymocytes did not form such rosettes. Spleen cells were passed over a Sephadex G-10 column or incubated on a plastic surface in order to eliminate adherent cells from them. Cells obtained by both these methods were unable to form rosettes. B cell-, T cell- and natural killer cell-enriched fractions in spleen cells were unable to form rosettes either. Some of the mouse IgG subclasses suppressed rosette formation when added to its site. These are monoclonal antibodies whose specificities are directed against Aspergillus niger glucose oxidase. Moreover, Aspergillus niger glucose oxidase suppressed the rosette formation when spleen cells had been treated with it as well as when it had been added to the site of rosette formation. These findings suggest that some murine spleen cells have receptors to a structure on autologous red blood cells, which is recognized by an anti-Aspergillus niger glucose oxidase monoclonal antibody. PMID- 10839566 TI - Intense eosinophilia with abnormal ultrastructure as presenting manifestation of acute lymphoblastic leukemia. AB - A patient with acute lymphoblastic leukemia presented with intense eosinophilia. Under the light microscope these eosinophils showed smaller eosinophilic granules and were detected as neutrophils by Coulter Gen-S cell counter. This counter identifies cell morphology by size and forward and right angle light scatter of cells. Under electron microscopy these eosinophils had smaller and fewer granules and very few crystalloid structures, thereby explaining the inability of the cell counter to identify them as eosinophils. Eosinophilia subsided at 6 months of treatment, i.e. 5 months after the patient went into morphological remission; cytogenetic and bone marrow analyses revealed no abnormality. PMID- 10839568 TI - No child should ever be hit. PMID- 10839567 TI - Primary malignant lymphoma of the central nervous system presenting with ascites and pleural effusion. AB - A 70-year-old woman was admitted to our hospital owing to ascites and pleural effusion. Though malignant cells (B-cell type lymphoma) were detected in both the ascites and pleural effusion, neither lymph node swelling nor a tumor was detected upon chest, abdominal and pelvic computed tomography (CT). After weekly THP-COP therapy for 8 weeks, the ascites and pleural effusion completely disappeared. Two years after the first admission, she was re-admitted because of a disturbance of consciousness, and a brain tumor was detected on CT scan. The immunohistological and genetic data for the brain tumor were identical to those of the malignant cells in the pleural effusion and ascites detected 2 years previously. Whereas the symptoms at onset of a primary lymphoma of the central nervous system (CNS) are usually neurological ones, in this rare case of primary CNS lymphoma, the symptoms at onset were the ascites and pleural effusion without neurological symptoms. PMID- 10839569 TI - Nutrition in pregnancy. PMID- 10839570 TI - Long-term effects of labor analgesia. PMID- 10839571 TI - Rediscovering the "M" in "MCH": maternal health promotion after childbirth. AB - Although maternal mortality is not a major health concern in the United States, evidence is accruing that after 6 weeks postpartum mothers continue to face mental and physical health, lifestyle, and parenting concerns. Exemplar areas for enhanced maternal health promotion after childbirth include (a) lifestyle changes in exercise, nutrition, and smoking, and (b) psychosocial well-being, particularly mood and body image. Research on health of women after childbirth supports rethinking the scope and duration of maternal health promotion. PMID- 10839572 TI - Ogilvie's syndrome after cesarean delivery. AB - Ogilvie's syndrome is a rare postsurgical complication that can be associated with cesarean delivery. It is characterized by massive dilation of the colon, much like that which occurs with an obstruction but in the absence of a mechanical obstruction. Early detection and intervention are necessary to avoid serious morbidity and/or mortality. Conservative treatment is effective in many cases, but surgical intervention may be required. Nursing assessment of the gastrointestinal system in the postsurgical patient is reviewed using a case report of a patient who developed Ogilvie's syndrome after a cesarean delivery. PMID- 10839573 TI - Mothers' performing creamatocrit measures in the NICU: accuracy, reactions, and cost. AB - OBJECTIVE: To determine whether mothers of infants in the neonatal intensive-care unit could be taught to perform creamatocrits (CRCTs) accurately on own mothers' milk (OMM). These mothers' reactions to performing CRCTs also were measured. SAMPLE: Twenty-six mothers and four advanced-practice nurses (RNs) participated in this study. DESIGN: For Phase One of this blinded trial, mothers were taught to perform the CRCT by one of the two instructional RNs. For Phase Two, mothers and one of the two validation RNs performed CRCTs simultaneously and independently on the same OMM sample, and the mother completed a Maternal Reactions questionnaire. RESULTS: Mothers' CRCT measures were highly accurate. The mean absolute difference between RNs' and mothers' CRCTs was 0.69%, with 50% and 84.6% of these differences, respectively, < or = 0.5% and < or = 1.0% CRCT. A strong linear correlation was noted between RNs' and mothers' CRCTs. Ninety-six percent of the mothers reported that the CRCT was easy to learn, they felt comfortable performing the procedure, and it made them feel more involved in infant care. A mean of 23.6 minutes was spent teaching the mother to perform CRCT, a figure that reflects the cost-effectiveness of the approach. CONCLUSION: Mothers can be taught to perform CRCTs accurately and easily on their OMM. This practice exemplifies high quality, cost-effective care that maximizes maternal involvement and satisfaction. PMID- 10839574 TI - Exercise during pregnancy and type of delivery in nulliparae. AB - OBJECTIVE: To assess the association between participation in aerobic exercise during the first two trimesters of pregnancy and type of delivery in nulliparous women. DESIGN: Nonexperimental, retrospective. SETTING: A large metropolitan area in the southwestern United States. PARTICIPANTS: 137 nulliparous women. OUTCOME MEASURES: Method of delivery. RESULTS: An unadjusted odds ratio showed that sedentary women (n = 93) were 2.05 times more likely to deliver via cesarean section than active women (n = 44), but this relationship was not statistically significant. Through logistic regression analysis with control for the mother's prepregnancy exercise program, age, use of epidural anesthesia, change in prepregnancy to delivery body mass index, labor length, whether labor was induced, and the hospital of birth, the odds of cesarean delivery were found to be 4.5 times greater for sedentary women than for active women. CONCLUSION: Regular participation in physical activity during the first two trimesters of pregnancy may be associated with reduced risk for cesarean delivery in nulliparous women. PMID- 10839575 TI - Teaching new mothers: priorities of nurses and postpartum women. AB - OBJECTIVE: To identify the most important content areas to include when teaching postpartum women, either in the hospital or at home, from the perspective of both nurses and postpartum women. DESIGN: Descriptive design using scaled response surveys. SETTING: Data for nurses were collected through mailed surveys. Data for new mothers were collected in the hospital within 24 hours after delivery and through mailed surveys. PARTICIPANTS: Seventy-one registered nurses who provided in-hospital maternity nursing care; 53 registered nurses who provided postpartum follow-up home care visits; and 103 low-risk postpartum women. MAIN OUTCOME MEASURES: Perceived importance of teaching topics related to care of mothers and their newborns. Comparisons between responses of nurses and new mothers were analyzed. RESULTS: Statistically significant differences were noted between the perceptions of nurses and new mothers regarding the priority of teaching content for new mothers. Nurses give priority to teaching about infant care, whereas new mothers give priority to their own care. CONCLUSIONS: These data suggest that new mothers may have opinions that differ from those of nurses as to what is important to learn about caring for themselves and for their newborns during the early postpartum period. PMID- 10839576 TI - Characteristics of Norplant users. AB - OBJECTIVE: To determine whether a profile can be developed to indicate which clients are the best candidates for Norplant by describing the characteristics of clients who use this contraceptive for 5 years and those who have it removed early. DESIGN: A descriptive study of data collected from a convenience sample. SELLING: A state-supported university hospital's obstetric and gynecology outpatient clinic serving primarily indigent clients in the southern United States. PARTICIPANTS: Women (N = 256), primarily with low incomes, being seen for removal of Norplant contraceptive capsules. Of these, 166 women had completed the recommended 5 years. MAIN OUTCOME MEASURES: Characteristics of the clients who retained the contraceptive for the recommended 5 years and characteristics of clients who came for early removal. RESULTS: Clients who retained the contraceptive for 5 years differed from those who did not on only three characteristics: race, age, and parity. No significant differences were found in education or marital status. CONCLUSIONS: No clear profile of successful Norplant users emerged. Instead, thorough counseling of each individual is essential to the client's completion of 5 years with Norplant. Advantages and disadvantages must be explained and the client's tolerance of possible side effects must be assessed, thereby helping the client to make an informed choice. PMID- 10839577 TI - Breastfeeding and adolescents. AB - OBJECTIVES: To critically review research on adolescent infant-feeding decision making and breastfeeding experiences. DATA SOURCES: Articles published between 1980 and 1999 on breastfeeding, infant feeding, and adolescence were located using CINAHL, MEDLINE, and PsycLit databases. STUDY SELECTION: English language research dealing with the subject was chosen. DATA EXTRACTION: Study findings were categorized: incidence of breastfeeding among adolescent mothers, physiologic aspects of lactation among teens, nonpregnant adolescents' breastfeeding knowledge and attitudes, factors related to infant-feeding decisions, and the adolescent breastfeeding experience. DATA SYNTHESIS: Most studies were descriptive in nature, with only two intervention studies published. Studies indicated that adolescents breastfeed less often than adults; teen mothers' breast milk is comparable to adults'; nonpregnant, pregnant, and delivered teens hold positive and negative attitudes toward breastfeeding that influence decision-making; and the breastfeeding teen has positive and negative experiences, some of which appear unique to her social situations and developmental level. CONCLUSIONS: There are considerable data on infant-feeding attitudes and decision-making among teens. Intervention research is indicated to improve breastfeeding initiation and duration. Further research is needed on actual infant feeding experiences of teens. Finally, descriptive research to identify health care professionals' beliefs and practices related to teens' choice and maintenance of breastfeeding is needed. PMID- 10839578 TI - History and development of fetal heart assessment: a composite. AB - Methods of assessing the fetal heart remained unchanged for approximately 150 years until the first commercial monitor suitable for clinical practice was sold in 1968. The impact and events of the last 30 to 40 years surrounding fetal heart assessment are revealed in perspectives of the past, present, and near future. Assessment practices have been shaped by the development of biotechnology, unrealistic expectations, interpretation disagreement, consumer response, and the practice and educational resources written by nursing and medicine. PMID- 10839579 TI - Fetal heart rate auscultation: current and future practice. AB - Intermittent auscultation (IA) has been reported as equivalent to electronic fetal monitoring (EFM) as a fetal surveillance method in terms of neonatal outcomes based on randomized controlled trials and meta-analyses. Despite recommendations to include IA as a primary method for fetal evaluation, EFM use predominates. Understanding the equipment, method, benefits and limitations, and strategies for implementing IA may assist nurses in providing informed choices for low-risk pregnant women. PMID- 10839580 TI - Retrofitting technology to nursing: the case of electronic fetal monitoring. AB - OBJECTIVES: To describe how nurses put electronic fetal monitoring to use in the 1960s and 1970s and the dilemmas this caused. DESIGN: Social history. RESULTS: Nurses used electronic fetal monitoring to improve the watchful and comfort care of childbearing women, and they saw it as validating nursing. They retrofitted, or worked to reconcile machine monitoring with natural, prepared, and participative childbirth, and with attentive and embodied nursing. CONCLUSION: Electronic fetal monitoring was another in a long line of technological innovations that fell to nurses to put into use and to make work. The remarkably rapid way electronic fetal monitoring became routine in the United States depended, in large part, on the articulation work of nurses. However, like all such work, what nurses did to make electronic fetal monitoring work for patients, physicians, hospitals, and manufacturers was largely invisible. Retrofitting efforts often entail unrecognized innovation and risks for nursing. PMID- 10839581 TI - Limited obstetric ultrasound in nursing practice. AB - During the past decade, nurses have begun to incorporate limited ultrasound into obstetric nursing practice. Guidelines have been established that provide the recommended content for the didactic and clinical preparation needed before implementing this technique. Fetal sonography provides useful information on fetal status and complements the fetal heart assessment. With appropriate education and institutional policies, nurses can enhance their role with this new skill. Acquiring an understanding of the educational and clinical guidelines for limited sonography is the first step for perinatal nurses who want to incorporate this skill into their clinical practice. PMID- 10839582 TI - Perioperative assessment of fetal heart rate and uterine activity. AB - Improvements in surgical techniques and anesthesia allow women the option to schedule needed surgery during pregnancy. However, perioperative monitoring of the fetus and uterine activity remains a matter of controversy. Monitoring may allow rapid improvement of the fetal status or uterine activity when early compromise or contractions are detected. The reassurance and decreased medicolegal risks provided by perioperative monitoring may offset the cost of a perinatal nurse and use of monitoring equipment even though the drug and anesthetic effect on the fetal heart limit the benefits of monitoring. Simply providing adequate maternal respiratory support during surgery may improve the fetal pattern but will not eliminate external surgical effects. The need for additional research is described, and the role of the perinatal nurse is detailed in a suggested protocol. PMID- 10839583 TI - Maximum suppression of HIV replication leads to the restoration of HIV-specific responses in early HIV disease. AB - OBJECTIVES: It is predicted that HIV-infected individuals in early HIV disease are the most likely group to achieve immune reconstitution following highly active antiretroviral treatment. We assessed whether suppression of HIV replication in this group would improve immune function. METHODS: Seventeen antiretroviral-naive patients in early HIV disease were evaluated for immune function and lymphocyte phenotyping using standard immunological assays. RESULTS: Absolute CD4+ T-cell number increased from a median of 550 to 800 x 10(6) cells/l while CD8+ T-cell numbers were reduced. The decrease in CD8+ cells correlated with a decrease in the CD8+ memory phenotype. Kinetics of CD4+ naive and memory T cell rise indicated that 80% of the maximum CD4+ naive increase was achieved within 18 weeks whereas maximum CD4+ memory T-cell rise was achieved within 36 weeks. Activation markers (HLA-DR, CD38) and an apoptosis-related marker (CD95) were reduced on CD4+ and CD8+ T cells. Lymphocyte proliferation responses to tetanus toxoid, alloantigen, and anti-CD3/CD28 were restored in patients that were initially unresponsive. At baseline, 31% of the patients responded to HIV p24, which increased to 69% post-therapy. The inducible RANTES response was normalized following therapy whereas inducible interferon-gamma, interleukin (IL) 12, and MIP1beta were elevated. The depressed inducible IL-10 response, however, was not altered after therapy. CONCLUSIONS: This is one of the first studies to demonstrate the restoration of HIV-1 specific responses in non-acute HIV infection, suggesting early intervention with potent antiretroviral therapy may reverse immune-mediated damage not seen with treated patients who have more advanced disease. PMID- 10839584 TI - Accelerated replicative senescence of the peripheral immune system induced by HIV infection. AB - OBJECTIVES: HIV induces rapid turnover of T lymphocytes but whether this leads to replicative senescence of CD4+ and CD8+ cells and contributes to AIDS symptoms is unclear. The aim of this study was to address this question by analyzing telomere length in blood cell populations as a measure of replicative history in a significant number of patients infected with HIV. DESIGN: Total peripheral blood mononuclear cells (PBMCs), CD4+ or CD8+ cells were isolated from blood collected from a total of 73 HIV patients and 27 controls. Samples were isolated to measure telomere length, telomerase activity and proliferative ability, and analyses were carried out in a blind experimental protocol. METHODS: PBMCs isolated on Ficoll Hypaque gradients were washed and prepared for additional fractionation into CD4+ and CD8+ cells using antibody-bound magnetic beads. Total PBMCs, CD4+ and CD8+ cells were used for cell cycle analysis, for telomerase activity assays and were measured for telomere length using the terminal restriction fragment assay. RESULTS: Telomere analyses in this study show a clear (P < 0.0001) inverse relationship between telomere length and progression of immunosuppression, with HIV infection resulting in a five-fold or greater acceleration of aging of the circulating PBMC component of the immune system. Patients who are 37 years old showed telomere lengths similar to uninfected 75-year-olds. Telomere loss correlated well with progression of AIDS and with reduced proliferative ability of patient PBMCs but was unrelated to telomerase activity. Mean telomere length was shorter in both CD4+ and CD8+ cells, with three-fold higher rates of telomere loss for CD8+ lymphocytes. CONCLUSIONS: These data provide strong support for the occurrence of accelerated replicative aging of the peripheral immune system, possibly resulting in a loss of T cells leading to AIDS symptoms. PMID- 10839585 TI - The role of abacavir (ABC, 1592) in antiretroviral therapy-experienced patients: results from a randomized, double-blind, trial. CNA3002 European Study Team. AB - OBJECTIVE: To compare the antiviral activity of abacavir (ABC) with stable background therapy (SBG) and SBG alone in antiretroviral therapy-experienced subjects as demonstrated by the proportion of subjects with plasma HIV-1 RNA < or = 400 copies/ml, plasma HIV-1 RNA and CD4 cell count profiles, and safety and tolerance of the two regimens over 16 weeks. DESIGN: One-hundred and eighty-five HIV-1 infected adults, with CD4 cell counts > or = 100 x 10(6)/l and plasma HIV-1 RNA of 400-50,000 copies/ml and who had received SBG therapy for at least 12 weeks, were randomized to receive ABC (300 mg twice daily) or placebo in a double blind, multi-centre study. METHODS: Antiretroviral activity was assessed by measuring changes in plasma HIV-1 RNA levels and CD4 cell counts. Genotypic and phenotypic resistance was determined at baseline and week 16. Evaluation of safety and tolerance was based on clinical adverse events and laboratory analyses. RESULTS: At week 16 significantly more subjects receiving ABC + SBG had plasma HIV-1 RNA < or = 400 copies/ml (36/92, 39%) than subjects receiving SBG alone (7/93, 8%; P < 0.001). A similar response was observed in both the lamivudine naive and lamivudine-experienced subjects. The presence of the M184V mutation did not preclude an antiviral response to ABC; 73% of subjects with the M184V mutation alone experienced a > or = 1.0 log10 copies/ml reduction in plasma HIV-1 RNA or had a value of < or = 400 copies/ml by week 16. CONCLUSIONS: ABC was generally well tolerated and exerted significant antiviral effect when added to combination antiretroviral therapy over 16 weeks. PMID- 10839586 TI - Salvage therapy with abacavir plus a non-nucleoside reverse transcriptase inhibitor and a protease inhibitor in heavily pre-treated HIV-1 infected patients. Swiss HIV Cohort Study. AB - BACKGROUND: Highly active antiretroviral therapy (HAART) may fail, especially in pre-treated patients. OBJECTIVE: To examine retrospectively whether heavily pre treated patients not responding to HAART at least once respond to a salvage therapy with abacavir, a nucleoside reverse transcriptase inhibitor (NRTI) plus a non-nucleoside analogue reverse transcriptase inhibitor (NNRTI) and one or two protease inhibitors (PI). PATIENTS: We retrospectively identified and analysed patients followed in the Swiss HIV Cohort Study with > 1000 HIV RNA copies/ml on HAART, naive to abacavir who were switched to abacavir plus one NNRTI (efavirenz or nevirapine) and one or two PI which had not been used in the previous HAART. RESULTS: Of 23 identified HIV-infected patients with four (median) therapy changes before salvage, 10 patients (43%) achieved a decrease of plasma HIV RNA > 0.5 log10 at 6 months of therapy. After 6 months only two patients had an HIV-1 RNA < 500 copies/ml, one of them < 50 copies/ml. Seven patients increased their CD4 cell counts by > 30% above baseline. Three patients, all with CD4 cell counts < 100 x 10(6)/l before salvage therapy had a > 30% decline in CD4 cell count. An extended number of resistance-associated mutations was found in almost all patients at baseline. One patient had two new AIDS-defining events. Five patients (22%) discontinued treatment because of side-effects, mainly occurrence of a rash. CONCLUSION: Salvage therapy in intensively pre-treated patients has a low virological success rate despite usage of abacavir and NNRTI. Nevertheless, this did not correlate with immunological and clinical course. This study emphasizes the difficulty of second-line treatment in HIV-1 infection and stresses the need for new compounds. PMID- 10839587 TI - Decreased exposure to saquinavir in HIV-1-infected patients after long-term antiretroviral therapy including ritonavir and saquinavir. AB - OBJECTIVE: To explore whether steady-state plasma pharmacokinetics of ritonavir and saquinavir change during long-term treatment in HIV-1-infected patients on antiretroviral treatment including ritonavir and saquinavir. METHODS: The pharmacokinetics of ritonavir and saquinavir were assessed during an 8-h period on two occasions in six HIV-1 infected patients on stable twice daily treatment with ritonavir 400 mg, saquinavir 400 mg and stavudine 40 mg with or without lamivudine 150 mg twice daily. RESULTS: The first study day was 4-12 months (median 7 months) after the start of the current regimen. The second study day was 9-15 months (median 10 months) later. No significant differences were observed for the ritonavir pharmacokinetics between the first and second study day. However, median change in plasma trough level of saquinavir between the two study days was -30% (range -79 to +11%; P = 0.06). Median change in maximum plasma concentration was -40% (range -62 to +34%; P = 0.09). The median change in area under the plasma concentration versus time curve over 0-8 h was -33% (range 53 to +21%; P = 0.06). CONCLUSION: The exposure to saquinavir decreased over time in HIV-infected patients on stable antiretroviral therapy. These data suggest that regular monitoring of plasma drug concentrations should become part of routine patient care even in apparently compliant patients. PMID- 10839588 TI - Risks and benefits of replacing protease inhibitors by nevirapine in HIV-infected subjects under long-term successful triple combination therapy. AB - OBJECTIVE: To analyse the safety and efficacy of replacing the protease inhibitor (PI) by nevirapine (NVP) in subjects experiencing a long-term control of virus replication under a triple PI-containing antiretroviral combination. DESIGN: Prospective evaluation of 138 HIV-positive subjects with plasma viral load below 50 HIV-RNA copies/ml for the last 6 months under a triple PI-containing regimen, who were randomly assigned to either replace the PI by NVP (n = 104) or continue on the same treatment (n = 34). METHODS: Viral load, CD4 count, lipid profile, body-shape features, and quality of life parameters were all assessed at the time of randomization and every 3 months thereafter. RESULTS: In an intent-to-treat analysis, a rebound in viral load occurred in 11% of subjects during the first 6 months after replacing the PI by NVP, whereas it appeared in 29% of those who remained on PI (P = 0.007). Treatment failure was related to lack of adherence in 90% of subjects on PI, but only in 22% of those receiving NVP (P = 0.006). The CD4 cell count outcome did not differ significantly comparing both groups at 6 months, although in patients receiving NVP an average reduction of 35 x 10(6) cells/l was observed, whereas in those on PI a positive trend was still recorded (+54 x 10(6) cells/l). At the time of randomization, 77.5 and 57.5% of subjects had cholesterol and triglyceride values above 200 mg/dl, respectively. No significant changes in the lipid profile were observed in any of the groups thereafter. Body-shape abnormalities were recorded in 70% of persons at the time of randomization, and partially reversed at 6 months in 50% of subjects who replaced the PI by NVP. A quality of life score recorded a significant improvement in subjects who switched to NVP compared with those who continued on PI. CONCLUSIONS: The replacement of PI by NVP seems to be safe both virologically and immunologically, provides a significant improvement in the quality of life and in half of patients ameliorate lipodystrophic body-shape changes at 6 months, although serum lipid abnormalities still remain unmodified. PMID- 10839589 TI - Viral genetic heterogeneity in HIV-1-infected individuals is associated with increasing use of HAART and higher viremia. AB - OBJECTIVE: To assess the correlation between the outgrowth of mutant viruses (viral genetic heterogeneity), highly active antiretroviral therapy (HAART), and plasma HIV-1 RNA in a population-based observational cohort study. DESIGN: The study population consisted of 42 HIV-1-infected individuals receiving at least two nucleotide reverse transcriptase (RT) inhibitors and one or more protease inhibitors at study entry. There were no restrictions on antiretroviral therapy after enrollment. METHODS: Plasma samples were obtained from subjects at baseline, at therapy changes, and at quarterly intervals for quantitation of HIV 1 RNA levels and for sequence determination of the entire protease coding region and the first 235 codons of the reverse transcriptase coding region. Data were analyzed using the generalized estimating equation method for longitudinal data and using linear regression analysis. RESULTS: With increased time on HAART there were significant increases in the number of total HIV-1 mutations in the regions sequenced (P = 0.010). There were significant correlations between the increases in the plasma HIV-1 RNA levels and the numbers of total mutations and reverse transcriptase mutations (P = 0.007 and 0.021, respectively). CONCLUSIONS: The number of HIV-1 mutations increased over time. Failure of HAART in this study population was correlated with outgrowth of virus with numerous mutations in the reverse transcriptase and protease coding regions. Phenotypic results correlated with genotypic results, showing decreased susceptibility to antiretrovirals over time in the majority of this population during HAART. Both synonymous and non synonymous mutations were observed, with a higher incidence of non-synonymous mutations occurring at codons associated with drug resistance. PMID- 10839590 TI - Effect of chemokine receptor gene polymorphisms on the response to potent antiretroviral therapy. AB - BACKGROUND: Both the natural history of HIV infection and the response to antiretroviral therapy are heterogeneous. Polymorphisms in chemokine receptor genes modulate the natural history of HIV-1 infection. In comparison with subjects with other genotypes, the prognosis for HIV-1-infected CCR5-delta32 heterozygotes is more favorable and that for CCR5 promoter allele 59029A homozygotes is less favorable. METHODS: HIV-1-infected adults with a CD4+ lymphocyte count > or = 200 cells x 10(6)/l and a plasma HIV RNA level > or = 1000 copies/ml were treated with indinavir, zidovudine and lamivudine for 6 months. HIV RNA levels were measured at 4-week intervals. Genotyping for chemokine receptor gene polymorphisms (CCR5-delta32, CCR5 59029A/G, CCR2-641) was performed. We examined whether the time to first HIV RNA < 200 copies/ml, frequency of viral suppression failure (HIV RNA > or = 200 copies/ml between weeks 16 and 28 of therapy), or reduction from the pre-treatment HIV RNA level differed by genotype. RESULTS: Time to first HIV RNA < 200 copies/ml was not predicted by genotype. Among 272 Caucasian patients, viral suppression failure was more common among patients with the CCR5 +/+ ? CCR2+/+ ? CCR5-59029 A/A genotype (28%) than among all other subjects combined (relative risk, 2.0; P = 0.06). After 24 weeks of therapy, genotype groups differed in the reduction of the HIV RNA level from baseline (P = 0.02); patients with the CCR5 +/+ ? CCR2+/+ ? CCR5-59029 A/A genotype had a mean reduction of 2.12 log10 copies/ml compared to 2.64 log10 copies/ml among all other groups combined. CONCLUSION: Polymorphisms in chemokine receptor genes may explain some of the heterogeneity in sustaining viral suppression observed among patients receiving potent antiretroviral therapy. PMID- 10839591 TI - Clinical course of cardiomyopathy in HIV-infected patients with or without encephalopathy related to the myocardial expression of tumour necrosis factor alpha and nitric oxide synthase. AB - OBJECTIVE: To define whether the development of encephalopathy influences the clinical course of HIV-associated cardiomyopathy (HIV-DCM) in relation to the myocardial expression of tumour necrosis factor-alpha (TNF-alpha) and inducible nitric oxide synthase (iNOS). DESIGN: Prospective study. SETTING: University hospitals and AIDS centres. METHODS: 115 HIV-infected patients with echocardiographic diagnosis of HIV-associated cardiomyopathy (34 with encephalopathy and 81 without encephalopathy) were followed for a mean of 24 +/- 3.2 months. All patients underwent endomyocardial biopsy for determination of myocardial immunostaining intensity of TNF-alpha and iNOS. Cerebrospinal fluid (CSF) from patients with encephalopathy was examined for the presence of viruses. Patients underwent clinical examination every 3 months and echocardiographic examination every 6 months. The intensity of TNF-alpha and iNOS immunostaining was also evaluated on postmortem cerebral tissue of patients who died of congestive heart failure (CHF). RESULTS: A greater impairment of echocardiographic parameters was observed in patients with HIV-associated cardiomyopathy after development of encephalopathy. These parameters tended to worsen progressively during the follow-up period and were inversely correlated with HIV-1 viral load, CD4 cell count, mini mental status score and the intensity of myocardial and cerebral TNF-alpha and iNOS staining. CSF specimens were available in 29 patients with encephalopathy. HIV-1 sequences were detected in CSF of all these patients with cytomegalovirus sequences in two. The mortality rate for CHF was greater among patients with encephalopathy (73% versus 12%). CONCLUSIONS: The development of encephalopathy has an adverse effect on the clinical course of HIV-associated cardiomyopathy. In the relationship between cardiomyopathy and encephalopathy, the activation of iNOS by TNF-alpha may have a significant pathogenetic role in HIV disease. PMID- 10839592 TI - Efficacy and safety of combination therapy with interferon-alpha2b and ribavirin for chronic hepatitis C in HIV-infected patients. AB - OBJECTIVES: To evaluate the efficacy and safety of a combination therapy of interferon-alpha2b (IFN) and ribavirin for the treatment of chronic hepatitis C in HIV-seropositive patients. DESIGN: Open prospective trial. METHODS: Twenty patients co-infected with hepatitis C virus (HCV) and HIV, with a mean CD4 cell count of 350 +/- 153 x 10(6)/l were treated with IFN (3 MU three times per week) in combination with ribavirin (500 mg or 600 mg twice a day) for 6 months. Tolerance and efficacy were monitored at weeks 12 (month 3) and 24 (month 6). The primary endpoint was a complete virological response, as defined by the lack of detectable HCV RNA in serum. RESULTS: Baseline values of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were 121 +/- 72 IU/l and 75 +/- 67 IU/l, respectively. The total Knodell score was 10.4 +/- 2.4, with nine patients showing histological evidence of active cirrhosis (45%). All patients exhibited circulating HCV RNA. The treatment was well tolerated, with no impact on the course of HIV infection. After 6 months of combination therapy with IFN and ribavirin, 10 patients (50%) exhibited no further detectable HCV RNA viraemia, seven of whom achieved undetectable viraemia at month 3. Levels of ALT and AST decreased after 6 months of treatment from a mean of 121 +/- 72 to 51 +/- 40 IU/l and from a mean of 129 +/- 58 IU/l to 68 +/- 61 IU/l, respectively (P < 0.0002 and P < 0.0001). CONCLUSION: Our results indicate that combination therapy with IFN and ribavirin is effective in 50% of cases in clearing serum HCV RNA and may thus provide effective means of therapy in HIV-HCV-coinfected patients as initial treatment or in patients who have previously failed IFN monotherapy. PMID- 10839593 TI - Amphotericin B oral suspension for fluconazole-refractory oral candidiasis in persons with HIV infection. Adult AIDS Clinical Trials Group Study Team 295. AB - OBJECTIVE: To determine the efficacy and safety of amphotericin B oral suspension (ABOS) for the treatment of fluconazole refractory oral candidiasis in persons with HIV infection. DESIGN AND SETTING: A prospective, multicenter, open label trial at 25 study centers within the AIDS Clinical Trials Group. PATIENTS AND METHODS: Individuals with diffuse oral candidiasis after 14 days of treatment with 200 mg of fluconazole daily (more than five plaques or a single plaque > 3 cm largest length) were treated with ABOS, 100 mg/ml, 5 ml swish and swallow, four times daily for 14 days. Thereafter incomplete or non-responders received an additional 14 days of therapy and responders received maintenance ABOS twice daily for up to 6 months. Relapses during maintenance ABOS were treated by increasing the dose to four times daily. MAIN OUTCOME MEASURES: To demonstrate an ABOS clinical response rate > 33% and a treatment-limiting toxicity rate < 50%. Clinical response was defined as the absence of mouth pain and the presence of less than five oral plaques, the largest being < 3 cm largest dimension. RESULTS: Fifty-eight subjects with a median age of 39 years and a median CD4 count of 10 x 10(6) cells/l were enrolled. Four subjects were excluded from the analysis because of inadequate follow-up after randomization (n = 3) or the presence of active esophageal disease (n = 1). Of the remaining 54 subjects, 23 (42.6%; 95% lower confidence interval, 31.1%) were classified as responders after 28 days. Five subjects (9%) stopped treatment due to toxicity. Relapse occurred in 16 responders (70%). CONCLUSIONS: Amphotericin B oral suspension is well tolerated but has limited efficacy for the treatment of fluconazole refractory oral candidiasis. PMID- 10839594 TI - CD4 cell counts in adults with newly diagnosed HIV infection: results of surveillance in England and Wales, 1990-1998. CD4 Surveillance Scheme Advisory Group. AB - OBJECTIVES: To describe the distribution and changes in CD4 cell counts (both initial and subsequent) in HIV-infected persons over time and determine the factors influencing these counts. DESIGN: Reports were requested from laboratories measuring CD4 cell counts in England and Wales. Initial counts were analysed and median counts were followed over time. METHODS: Time trends and the relationship between initial CD4 cell count and age, sex, and HIV risk category were studied using quantile regression methods or chi-square tests. RESULTS: Between 1990 and 1998, 9553 adults were newly diagnosed with HIV infection and had a CD4 cell count within 6 months of HIV diagnosis. Over 50% of initial CD4 cell counts in each major risk category were below 350 cells/mm3. Older age (P < 0.001), male sex (P < 0.013) and heterosexual risk (P < 0.001) were independently associated with lower initial CD4 cell counts. For heterosexually infected adults, the median initial CD4 cell count was significantly negatively associated with the year of diagnosis (P = 0.03) and the median age increased through the time period examined (P < 0.001), whereas for men who have sex with men (MSM), there was no significant change in these values over time. For each year cohort of newly diagnosed individuals, the median CD4 cell count in subsequent years decreased until 1996 and then increased thereafter, consistent with a treatment effect. CONCLUSION: Across all major risk groups, a large proportion of HIV infected adults are being diagnosed late in the course of HIV disease. For the heterosexually infected, the data suggest an ageing cohort effect, whereas for MSM the data are consistent with continuing transmission. PMID- 10839595 TI - Serum albumin as a predictor of survival in HIV-infected women in the Women's Interagency HIV study. AB - BACKGROUND: The level of serum albumin is associated with mortality in a wide variety of chronic diseases. However, few studies have examined the relationship between serum albumin and survival in HIV-1 infection. OBJECTIVES: To determine whether the serum albumin level is associated with survival in HIV-1 infected women. DESIGN: Prospective cohort study. Patients were interviewed and examined at 6 month intervals. SETTING: A North American multi-institutional cohort of HIV infected women from five geographical areas. PARTICIPANTS: A total of 2056 HIV infected women at various stages of disease. MEASUREMENTS: Mortality during the first 3 years of follow-up. The relative risk of death by serum albumin level was estimated using a proportional hazards ratio adjusted for CD4 cell count, HIV-1 RNA level and other relevant covariates. RESULT: Three year mortality for women in the lowest serum albumin category (< 35 g/l) was 48% compared with 11% in the highest category (> or = 42 g/l; P < 0.001). The adjusted relative hazard (RH) of death was 3.1 times greater for those in the lowest albumin category (P < 0.01). The excess risk associated with lower serum albumin levels remained when subjects with moderate to severe immunosuppression and abnormal kidney and liver function were excluded (P < 0.01). CONCLUSION: The baseline serum albumin level is an independent predictor of mortality in HIV-1-infected women. The serum albumin level may be a useful additional marker of HIV-1 disease progression, particularly among asymptomatic women with little or no evidence of immunosuppression. PMID- 10839596 TI - Trends in HIV seroprevalence among patients with sexually transmitted diseases in 17 European sentinel networks, 1990-1996. European Study Group. AB - OBJECTIVE: To monitor trends in HIV seroprevalence among sentinel populations of patients with new sexually transmitted disease episodes in 17 networks of 15 European countries. METHODS: Time trends were estimated by logistic regression for homo-/bisexual men, injecting drug users (IDU) and non-IDU heterosexuals. The networks were included as independent variables to account for different prevalence levels across Europe. Socio-demographic and behavioural data were also included in the model. The interactions of time and each factor in the model were evaluated to determine whether trends differed across networks or population subgroups. RESULTS: Overall, more than 150,000 sexually transmitted disease episodes were registered in this study. In the European network as a whole, the prevalence decreased significantly among IDU [n = 2619; odds ratio (OR) for annual change, 0.86; 95% confidence interval (CI), 0.80-0.93]. It also decreased among homo-/bisexual men (n = 11,809; OR, 0.92; 95% CI, 0.89-0.94). In both groups, trends did not differ statistically between networks. Among non-IDU heterosexuals (n = 114,024) the prevalence increased significantly (OR, 1.08; 95% CI, 1.04-1.13), especially among women (OR, 1.13) and the time trends differed statistically across networks. A significant increase was observed in four networks, whereas no specific change was detected in the others. CONCLUSIONS: By applying a standardized protocol, trends in HIV prevalence could be compared across networks and estimated at a more global level. For the validity of HIV trends in such surveys, it is essential to minimize the number of patients not being tested in networks where voluntary testing is required. PMID- 10839597 TI - A decade of HIV/AIDS prevention in Sweden: changes in attitudes associated with HIV and sexual risk behaviour from 1987 to 1997. AB - OBJECTIVE: The objective of the present study was to monitor the changes in determinants of HIV-related attitudes and behaviour over a 10 year period. DESIGN/METHODS: In the past decade, over 11,000 individuals in Sweden participated in this repeated cross-sectional study to assess the changes in attitudes and behaviour associated with HIV. Changes in the fear of persons who are HIV seropositive, optimism that scientists will find a cure or vaccine for HIV/AIDS, engagement in discussions concerning HIV/AIDS as well as condom use, the number of sexual partners, and frequency of unprotected casual sex were assessed during four study years, 1987, 1989, 1994, and 1997. RESULTS: The findings of this study suggest that changes in attitudes regarding HIV were more robust whereas only modest changes in sexual behaviour were observed. The fear of HIV-seropositive persons, optimism that a cure or vaccine would be developed, and engagement in discussions concerning HIV all decreased over the 10 year period. A trend in increased condom use was observed, particularly in younger participants, but little change in sex with multiple partners and unprotected casual sex was observed between 1987 and 1997. CONCLUSION: Changes in attitudes regarding HIV are usually more often observed than changes in sexual behaviour. However, increased condom use in younger participants may be reflective of secular changes in sexual behaviour that may facilitate the prevention of HIV in Sweden. PMID- 10839598 TI - CD8 T cell anti-HIV activity as a complementary protective mechanism in vaccinated chimpanzees. PMID- 10839599 TI - Stromal-derived factor-1 chemokine gene variant is associated with the delay of HIV-1 disease progression in two longitudinal cohorts. PMID- 10839600 TI - HIV transmission between two siblings in Africa. PMID- 10839601 TI - Widespread circulation of a B/F intersubtype recombinant form among HIV-1 infected individuals in Buenos Aires, Argentina. PMID- 10839602 TI - Valproic acid induces human herpesvirus 8 lytic gene expression in BCBL-1 cells. PMID- 10839603 TI - Antiretroviral drug removal by haemodialysis. PMID- 10839604 TI - Protease inhibitor therapy-associated lipodystrophy, hypertriglyceridaemia and diabetes mellitus. PMID- 10839605 TI - Lipodystrophy and long-term therapy with nucleoside reverse transcriptase inhibitors. PMID- 10839606 TI - Detection of drug resistance associated mutations in HIV primary infection within the UK. PMID- 10839607 TI - Maternal vitamin A status and mother-to-child transmission of HIV in West Africa. DITRAME Study Group. PMID- 10839608 TI - Polymerase chain reaction on sputum for the diagnosis of pulmonary toxoplasmosis in AIDS patients. PMID- 10839609 TI - Mixed endocrine-exocrine tumors of the gastrointestinal tract. AB - Mixed endocrine-exocrine tumors of the gastrointestinal tract are rare neoplasms, which have been reported in the literature mainly as case reports and have been designated with a various and rather confusing terminology. In this review, on the basis of personally studied cases and of the analysis of cases reported in the literature, we have tried to identify types of mixed endocrine-exocrine tumors showing different clinicopathologic and biological characteristics. We have also tried to group the different clinicopathologic entities in prognostic classes which include: benign, low-grade, intermediate grade, and high-grade malignant mixed endocrine-exocrine tumors. The criteria for identifying the various types of mixed endocrine-exocrine tumors are extensively discussed. PMID- 10839610 TI - Mixed exocrine-endocrine tumors of the pancreas. AB - Neoplasms of the pancreas usually show either ductal, acinar, or endocrine differentiation. Mixed exocrine-endocrine pancreatic neoplasms, in which the endocrine component is significant and comprises one-third to one-half of the tumor tissue, are rare. Truly mixed tumors have to be distinguished from exocrine neoplasms with scattered endocrine cells. In ductal adenocarcinomas, the scattered endocrine cells seem to be nonneoplastic. In other malignancies such as acinar cell carcinoma and pancreatoblastoma, scattered endocrine cells most likely represent an integral component of the tumor. PMID- 10839611 TI - Thyroid carcinomas with mixed follicular and C-cell differentiation patterns. AB - Divergent endocrine-neuroendocrine differentiation in thyroid carcinoma occurs in mixed medullary-follicular carcinomas (MMFC). Less than 40 cases of MMFC have been reported having highly heterogeneous patterns of growth. Classical medullary carcinoma areas may be intermingled with follicles or papillae or even oxyphilic and solid areas. Calcitonin and thyroglobulin are expressed in different cell populations. Presence of the latter suggests a potential usefulness of radioiodine treatment. The clinical behavior of MMFC does not differ from that of ordinary medullary carcinoma. The histogenesis of MMFC is controversial. The genetic analysis of the 2 neoplastic components showed that they are not derived from a common precursor, but rather display remarkable differences in the genetic profile (RET mutations and allelic losses). In addition, in some cases the follicular component was found to be oligo/polyclonal and therefore possibly hyperplastic rather than neoplastic. The follicular cells may have grown into the medullary carcinoma, after acquiring some molecular defect, being "hostage" of the true neoplastic (medullary) component. PMID- 10839612 TI - Divergent differentiation in neuroendocrine tumors of the adrenal gland. AB - Composite tumors of the adrenal medulla usually consist of pheochromocytoma admixed with ganglioneuroma or ganglioneuroblastoma. These neoplasms reflect phenotypic plasticity shown by primitive sympathetic cells and mature chromaffin cells in vitro. They may give rise to metastatic neuroblastoma in adults and may cause signs and symptoms attributable to both catecholamine and neuropeptide production. Schwann cells and sustentacular cells are typically numerous in these tumors but it is not known whether they are neoplastic. Immunohistochemical staining for catecholamine biosynthetic enzymes, secretory vesicle proteins and S 100 protein tends to recapitulate staining of the normal adrenal medulla or sympathetic ganglia. Sparsity of chromogranin A in the cell bodies of immature and mature neurons is a diagnostically useful characteristic. PMID- 10839613 TI - Expression of the neuroendocrine phenotype in carcinomas of the breast. AB - Neuroendocrine (NE) features are detectable in carcinomas of the breast either as scattered cells immunoreactive for NE markers in carcinoma of the usual type (NOS), or as special type of tumors where the vast majority of the cells display NE characteristics. The former type of lesions, whose biological and diagnostic significance is not clear yet, might reproduce the same phenomenon known to occur in carcinomas of the gastrointestinal tract and pancreas. In the present review we focus on the latter type of lesions, a spectrum of breast tumors largely composed of NE cells. These carcinomas, that we consider the "NE differentiated carcinomas of the breast," are here distinguished from "breast carcinomas NOS with NE differentiation." The diagnostic and histogenetic features of the various types of "NE differentiated carcinomas of the breast," their histological and cytological features and the role and value of ancillary diagnostic techniques, are reviewed. Data of the literature are discussed and related to a relatively large personal series. In addition, divergent differentiation in NE carcinomas of the breast, which is a relatively frequent phenomenon of diagnostic interest but of unknown significance (mainly involving mucinous intra- and extracellular production) is discussed. PMID- 10839614 TI - Divergent differentiation in neuroendocrine lung tumors. AB - The classification of neuroendocrine (NE) lung tumors has been revised in the 1999 World Health Organization (WHO) classification of lung tumors, allowing sharp morphological definition of typical versus atypical carcinoids, and atypical carcinoids versus large cell neuroendocrine carcinoma (LCNEC), a newly described class of high-grade NE lung tumors which differs from small-cell lung cancer by a large-cell phenotype. Divergent differentiation accounts for the high frequency of glandular differentiation with mucin production, and ultrastructural features in carcinoids and LCNEC, and low frequency of squamous differentiation in both LCNEC and SCLC. Specific NE markers (chromogranin, synaptophysin, neural cell adhesion molecule) and epithelial markers consistently negative in neuroendocrine components (cytokeratins 1, 5, 10, 14; epidermal growth factor (EGF)-receptor, human leukocyte antigen beta 2 (HLA-beta2) microglobuline) help to recognize divergent differentiation in NE tumors. At morphological level, divergent differentiation in NE tumors is recognized in WHO classification as variants: combined SCLC and combined LCNEC. The derivation of all lung tumors from a common endodermal stem cell and adoption of amine precursor uptake and decarboxylation properties by this endodermal stem cell explains divergent differentiation in NE lung tumors and the occurrence of NE subsets in NSCLC. PMID- 10839615 TI - Divergent neuroendocrine differentiation in prostatic carcinoma. AB - A rich variety of neuroendocrine cells are present in the normal prostate gland. Prostatic carcinoma may show divergent differentiation towards a neuroendocrine phenotype in the form of neuroendocrine small cell carcinoma or carcinoid-like tumors. Much more common is focal neuroendocrine differentiation in prostatic adenocarcinoma which may be pronounced in approximately 10% of adenocarcinomas. The prognostic significance of focal neuroendocrine differentiation in prostatic carcinoma is controversial but current evidence suggests an influence on prognosis related to hormone resistant tumors and/or a role in the conversion to a hormonal resistant phenotype. Chromogranin A appears to be the best overall tissue and serum marker of neuroendocrine differentiation. Chromogranin A serum levels may be useful in the assessment of the emergence of and/or progression of hormone resistant cancer. PMID- 10839616 TI - Divergent differentiation in endocrine and nonendocrine tumors of the skin. AB - In the skin, endocrine tumors showing areas with nonendocrine features and nonendocrine tumors showing endocrine differentiation are present. (1) Neuroendocrine carcinomas with nonendocrine differentiation: Merkel cell carcinoma (MCC) of the skin has been frequently described in association with squamous cells carcinoma (SCC) which can arise separately (as synchronous or metachronous lesions) from MCC as well as closely intermixed. In the first event the possibility that the lesions are sustained by same causative factors (among which sun exposure is the most probable) is suggested. In cases of lesions closely intermixed the possibility of an origin from a common precursor is suggested. Furthermore, cases of MCC have been described to contain glandular, melanocytic, striated muscle, and lymphoepithelioma-like features. These latter findings further support the hypothesis of tumors showing divergent differentiations. (2) Nonendocrine tumors showing endocrine differentiation: Basal cell carcinoma (BCC) was the first cutaneous nonendocrine tumor described to contain neuroendocrine granules. Presence of endocrine features were subsequently confirmed with immunohistochemical studies. Endocrine features were then described in sweat gland apocrine and eccrine carcinomas. Endocrine elements present in BCC and in sweat gland carcinomas do not show morphological and immunohistochemical features of Merkel cells. Thus the possibility that these tumors develop an immature Merkel cell or a new type of endocrine cell of the skin is suggested. Tumors with follicular differentiation such as trichoblastomas and trichofolliculomas contain a high number of Merkel cells. As Merkel cells are numerous in hair follicles of human fetal skin, the possibility that these tumors recapitulate the human skin embryogenesis is suggested. PMID- 10839617 TI - Utilization of the Streptoalloteichus hindustanus resistance determinant ShBle as a protein framework: effect of mutation upon ShBle dimerization and interaction of C-terminal displayed peptide epitopes. AB - We have selected the Streptoalloteichus hindustanus bleomycin-resistance protein ShBle, a 28-kDa homodimer, as a scaffold for the display of bioactive peptides and other peptide epitopes. To create a monomeric scaffold, we investigated the effect of mutating residue proline 9 to glycine. This residue plays a critical role in ShBle dimerization by affecting the position of the eight N-terminal residues which secure the interaction between the monomeric subunits. We demonstrate that this mutation weakens the dimerization interaction, resulting in establishment of a stable equilibrium between monomeric and dimeric ShBle species in solution. Circular dichroism and SDS-PAGE data indicate that the Pro9Gly mutation does not disrupt the structure of the molecule. Production of a fully monomeric form of ShBle required complete removal of the eight-residue N-terminal peptide, and the interaction across the now solvent-exposed hydrophobic interface of the ShBle monomer was insufficient to drive dimerization. To demonstrate efficient display of epitope tags on the ShBle protein, we displayed dual octapeptide FLAG tags at the protein C-terminus. These additions did not interfere with protein folding or activity. The resulting ShBle scaffold was used to compare the efficiency of two commercial FLAG-specific antibodies by biosensor. PMID- 10839618 TI - Fluorescence quenching studies of Trp repressor-operator interaction. AB - Steady-state quenching and time-resolved fluorescence measurements of L tryptophan binding to the tryptophan-free mutant W19/99F of the tryptophan repressor of Escherichia coli have been used to observe the coreperessor microenvirnment changes upon ligand binding. Using iodide and acrylamide as quenchers, we have resolved the emission spectra of the corepressor into two components. The bluer component of L-tryptophan buried in the holorepressor exhibits a maximum of the fluorescence emission at 336 nm and can be characterized by a Stern-Volmer quenching constant equal to about 2.0-2.3 M(-1). The second, redder component is exposed to the solvent and possesses the fluorescence emission and Stern-Volmer quenching constant characteristic of L tryptophan in the solvent. When the Trp holorepressor is bound to the DNA operator, further alterations in the corepressor fluorescence are observed. Acrylamide quenching experiments indicate that the Stern-Volmer quenching constant of the buried component of the corepressor decreases drastically to a value of 0.56 M(-1). The fluorescence lifetimes of L-tryptophan in a complex with Trp repressor decrease substantially upon binding to DNA, which indicates a dynamic mechanism of the quenching process. PMID- 10839619 TI - The cDNA-derived amino acid sequence of hemoglobin I from Lucina pectinata. AB - The tropical clam Lucina pectinata contains a unique hemoglobin (HbI) which serves to transport H2S to autotrophic bacteria. The cDNA-derived amino acid sequence was obtained from overlapping clones containing the cDNA that codes for HbI. The reverse transcriptase-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) methods were employed to synthesize the cDNA fragments. An initial 354-bp cDNA clone encoding 118 amino acid residues of HbI was amplified from total RNA by RT-PCR using degenerate oligonucleotides. Gene specific primers derived from the HbI-partial cDNA sequence were used for obtaining the 5' and 3' ends of the cDNA by RACE. The length of the HbI cDNA, estimated from sequence analysis of overlapping clones, was 1322 bp for the full length cDNA. The coding region of the full-length cDNA codes for 143 amino acid residues. The most conserved amino acid residues in HbI from Lucina pectinata were identified by a multiple alignment with nonvertebrate globin sequences. PMID- 10839620 TI - Identification and partial purification of DnaK homologue from extremely halophilic archaebacteria, Halobacterium cutirubrum. AB - The levels of synthesis of six proteins were increased at elevated growth temperature of the extremely halophilic archaebacterium Halobacterium cutirubrum. One of these proteins, with an apparent molecular mass of 97 kDa on sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE), bound to an ATP agarose column in the presence of 4 M NaCl, but not in the absence of salt, indicating that this protein retained its ATP-binding activity only at high salt concentration. The NH2-terminal sequence of this protein and the internal sequences of the tryptic peptides covering 1/3 of the total number of residues coincided with that deduced from the nucleotide sequence of the dnaK gene isolated from H. cutirubrum. The results strongly suggest that this apparent 97 kDa protein is the gene product of dnaK, although the molecular mass calculated from the nucleotide sequence is only 68,495, much smaller than the value of this protein determined by SDS-PAGE. Ferguson plot analysis indicated that this protein showed anomalous mobility on SDS-PAGE. We have purified DnaK homologue to greater than 90% homogeneity with stepwise elution from an ATP-agarose column. PMID- 10839621 TI - Nucleic acid-binding properties of low-density lipoproteins: LDL as a natural gene vector. AB - The role of apo B-100 as a transcription factor is indicated by the presence of regions in its primary structure that are similar to the DNA-binding domains of the transcription factors ISGF3gamma, STATs, IRFs, and SREBPs as well as by the presence of 11 RNA-binding KH domains. The Apo B-100 sequence also contains numerous bipartite nuclear localization sequences (NLS). A modified gel shift assay was used to show binding of highly purified preparations of human LDLs to fragmented genomic DNA, plasmid DNA, synthetic oligonucleotides (ISRE, 5' GGGAAACCGAAACTG and E/C, E-box motif and CCAAT, adipocyte-specific genes promoter site), and total RNA from human liver. LDL was observed to bind preferentially to plasmid DNA containing the hCMV IE2 promoter region. In experiments using human liver total RNA, RNA for five different genes was recovered from LDL and VLDL bands. Gene transfection experiments using human skin fibroblast cells were used to study the gene transfer capacity of LDL. Cells transfected with a pEGFP-N1 plasmid DNA and LDL expressed functional GFP, as indicated by fluorescence, at approximately 3 hrs after transfection. Our results strongly support an alternative role for apo B-100, in toto or perhaps as functional fragments, in the control of gene expression and as gene transfer vector. PMID- 10839622 TI - Identification of a glutathione-S-transferase effector domain for inhibition of jun kinase, by molecular dynamics. AB - We have recently found that the glutathione-S-transferase pi-isozyme (GST-pi), a cellular detoxification enzyme, potently and selectively inhibits activation of jun protein by its upstream kinase, jun kinase (JNK). This newly identified regulatory activity of GST-pi is strongly inhibited by a group of agents that inhibit its enzymatic activity. Since loss of enzymatic activity in general does not correlate with loss of regulatory activity, it is likely that inhibitor binding induces changes in the structure of one or more domains of GST that block its interaction with JNK. To identify regions of GST that change conformation on the binding of inhibitors, we have performed molecular dynamics calculations on GST-pi to compute its average structure in the presence and absence of the inhibitor, glutathione sulfonate. Superposition of the two average structures reveals that several regions change local structure depending upon whether the inhibitor is bound or not bound. Two of these regions, residues 36-50 and 194 201, are highly exposed. We have synthesized peptides corresponding to these two segments and find that the 194-201 sequence strongly inhibits the ability of GST pi to block the in vitro phosphorylation of jun by JNK. These results suggest that this region of GST-pi is critical to its functioning as a newly discovered regulator of signal transduction. PMID- 10839623 TI - Molecular dynamics analysis of the structures of ras-guanine nucleotide exchange protein (SOS) bound to wild-type and oncogenic ras-p21. Identification of effector domains of SOS. AB - The X-ray crystal structure of the ras oncogene-encoded p21 protein bound to SOS, the guanine nucleotide exchange-promoting protein, has been determined. We have undertaken to determine if there are differences between the three-dimensional structures of SOS bound to normal and oncogenic (Val 12-p21) proteins. Using molecular dynamics, we have computed the average structures for both complexes and superimposed them. We find four domains of SOS that differ markedly in structure: 631-641, 676-691, 718-729, and 994-1004. Peptides corresponding to these sequences have been synthesized and found to be powerful modulators of oncogenic p21 in cells as described in an accompanying paper. We find that the SOS segment from 809-815 makes contacts with multiple domains of ras-p21 and can facilitate correlated conformational changes in these domains. PMID- 10839624 TI - Inhibition of oncogenic and activated wild-type ras-p21 protein-induced oocyte maturation by peptides from the guanine-nucleotide exchange protein, SOS, identified from molecular dynamics calculations. Selective inhibition of oncogenic ras-p21. AB - In the preceding paper we performed molecular dynamics calculations of the average structures of the SOS protein bound to wild-type and oncogenic ras-p21. Based on these calculations, we have identified four major domains of the SOS protein, consisting of residues 631-641, 676-691, 718-729, and 994-1004, which differ in structure between the two complexes. We have now microinjected synthetic peptides corresponding to each of these domains into Xenopus laevis oocytes either together with oncogenic (Val 12)-p21 or into oocytes subsequently incubated with insulin. We find that the first three peptides inhibit both oncogenic and wild-type p21-induced oocyte maturation, while the last peptide much more strongly inhibits oncogenic p21 protein-induced oocyte maturation. These results suggest that each identified SOS region is involved in ras stimulated signal transduction and that the 994-1004 domain is involved uniquely with oncogenic ras-p21 signaling. PMID- 10839625 TI - Identification of the site of inhibition of oncogenic ras-p21-induced signal transduction by a peptide from a ras effector domain. AB - We have previously found that a peptide corresponding to residues 35-47 of the ras-p21 protein, from its switch 1 effector domain region, strongly inhibits oocyte maturation induced by oncogenic p21, but not by insulin-activated cellular wild-type p21. Another ras-p21 peptide corresponding to residues 96-110 that blocks ras-jun and jun kinase (JNK) interactions exhibits a similar pattern of inhibition. We have also found that c-raf strongly induces oocyte maturation and that dominant negative c-raf strongly blocks oncogenic p21-induced oocyte maturation. We now find that the p21 35-47, but not the 96-110, peptide completely blocks c-raf-induced maturation. This finding suggests that the 35-47 peptide blocks oncogenic ras at the level of raf; that activated normal and oncogenic ras-p21 have differing requirements for raf-dependent signaling; and that the two oncogenic-ras-selective inhibitory peptides, 35-47 and 96-110, act at two different critical downstream sites, the former at raf the latter at JNK/jun, both of which are required for oncogenic ras-p21 signaling. PMID- 10839626 TI - Dichain structure of botulinum neurotoxin: identification of cleavage sites in types C, D, and F neurotoxin molecules. AB - Botulinum neurotoxin (NT) is synthesized by Clostridium botulinum as about a 150 kDa single-chain polypeptide. Posttranslational modification by bacterial or exogenous proteases yielded dichain structure which formed a disulfide loop connecting a 50-kDa light chain (Lc) and 100-kDa heavy chain (Hc). We determined amino acid sequences around cleavage sites in the loop region of botulinum NTs produced by type C strain Stockholm, type D strain CB16, and type F strain Oslo by analysis of the C-terminal sequence of Lc and the N-terminal sequence of Hc. Cleavage was found at one or two sites at Arg444/Ser445 and Lys449/Thr450 for type C, and Lys442/Asn443 and Arg445/Asp446 for type D, respectively. In culture fluid of mildly proteolytic strains of type C and D, therefore, NT exists as a mixture of at least three forms of nicked dichain molecules. The NT of type F proteolytic strain Oslo showed the Arg435 as a C-terminal residue of Lc and Ala440 as an N-terminal residue of Hc, indicating that the bacterial protease cuts twice (Arg435/Lys436 and Lys439/Ala440), with excision of four amino acid residues. The location of cleavage and number of amino acid residue excisions in the loop region could be explained by the degree of exposure of amino acid residues on the surface of the molecule, which was predicted as surface probability from the amino acid sequence. In addition, the observed correlation may also be adapted to the cleavage sites of the other botulinum toxin types, A, B, E, and G. PMID- 10839627 TI - Lysine: N6-hydroxylase: stability and interaction with ligands. AB - Recombinant lysine:N6-hydroxylase, rIucD, which is isolated as an apoenzyme, requires FAD and NADPH for its catalytic function. rIucD preparations have been found to undergo time-dependent loss in monooxygenase function due to aggregation from the initial tetrameric state to a polytetrameric form(s), a process which is reversible by treatment with thiols. Ligand-induced conformational changes in rIucD were assessed by monitoring its CD spectra, DSC profile, and susceptibility to both endo- as well as exopeptidases. The first two methods indicated the absence of any significant conformational change in rIucD, while the last approach revealed that FAD, and its analog ADP, can protect the protein from the deleterious action of proteases. NADPH was partially effective and L-lysine was ineffective in this regard. Deletion of the C-terminal segment, either by treatment with carboxypeptidase Y or by mutagenesis of iucD, results in the loss of rIucD's monooxygenase activity. These findings demonstrate the crucial role of the C-terminal segment in maintaining rIucD in its native conformation. PMID- 10839628 TI - Glut-1 expression and its response to hypoglycemia in the embryonic mouse heart. AB - The embryonic heart depends on glucose during early organogenesis. Glut-1 functions in constitutive glucose uptake in adult tissues and is the predominant glucose transporter in embryonic and fetal tissues. This study focuses on Glut-1 expression in the heart during normal organogenesis using immunohistochemistry for Glut-1 distribution, Western analysis for Glut-1 protein levels, and reverse transcriptase polymerase chain reaction for Glut-1 mRNA levels. The role of Glut in glucose uptake response to hypoglycemia in the embryonic heart is evaluated using the Glut inhibitor cytochalasin B. Cardiac Glut-1 expression is also evaluated after in vitro hypoglycemic exposure. Glut-1 levels are highest on gestational days 9-10, intermediate on gestational day 10.5, and lowest on gestational days 11.5-13.5 in the normal embryonic heart. Cardiac Glut-1 mRNA levels similarly decline between gestational days 9.5 and gd 13.5. Cytochalasin B produces a dose-dependent decrease in glucose uptake in hearts exposed to hypoglycemia for 30 min or 6 h, implicating Glut in this response. Glut-1 protein expression is unchanged after 2 or 6 h but increased after 12 and 24 h of hypoglycemia in the gestational day 9.5 heart. Thus, Glut-1 expression is prominent in the embryonic heart and is correlated with changes in cardiac glucose requirements during normal organogenesis. Glut activity increases in response to acute hypoglycemia and the expression of Glut-1 increases in response to prolonged hypoglycemia. These results support the importance of Glut-1 during normal cardiogenesis and in response to hypoglycemia in the embryonic heart. PMID- 10839629 TI - Expression of 3 beta-hydroxysteroid/dehydrogenase/delta5-delta4/isomerase in the tracheal cartilage of the rat. AB - The enzyme complex 3beta-hydroxysteroid dehydrogenase isomerase/delta5-delta4 (3beta-HSD) is involved in the biosynthesis of all classes of active steroids. In this study, the presence of 3beta-HSD was defined in rat tracheal cartilage. The expression of the 3beta-HSD gene was examined by Northern blot analysis from 30 day-old rats. Western blot and immunohistochemical localization were also performed with antibodies raised against purified human placental 3beta-HSD to obtain further information on the expression of 3beta-HSD protein during fetal and postnatal periods of development in rat cartilage. Northern blot analysis using an oligonucleotide common to the 4 known 3beta-HSD isoforms showed 3beta HSD mRNA corresponding to a transcript of 1.7 kb. Furthermore, a 42 KDa protein band was detected in the tracheal cartilage extracts by Western blot analysis. Immunostaining for 3beta-SD was observed in chondrocytes. The first expression was detected on the 17th day of fetal life by Western blot and immunohistochemistry. Immunoreactivity of 3beta-HSD showed a significant increase at 7 and 15 days after birth, and then remained unchanged through adulthood, in agreement with the data of the Western blot. Our results demonstrated the expression for 3beta-HSD in the tracheal cartilage at both the mRNA and protein levels during fetal life and postnatal development of the rat. These results suggest that 3beta-HSD may synthesize certain steroids which play major roles in differentiation and maintenance of function during development of rat cartilage. PMID- 10839630 TI - Cutaneous glands in the Australian hylid Litoria caerulea (Amphibia, Hylidae). AB - Ultrastructure of cutaneous glands is described in the Australian hylid Litoria caerulea. Three main types of glands could be distinguished in both ventral and dorsal skin: mucous, serous or granular, and lipid glands. Both mucous, and to some extent, serous glands show a PAS-positive reaction. Some of the granular serous glands react to lipid staining. In addition, a very large gland confined to the dorsal skin of the head reacts to lipid staining. Apparently more than one type of dermal gland is involved in lipid secretion. The subject of skin lipid secretion is discussed in relation to the ecophysiological adaptations of this xeric-inhabiting frog. PMID- 10839631 TI - Expression of proto-oncogenes in bovine preimplantation blastocysts. AB - Proto-oncogenes are involved in the regulation of gene expression, for example after ligand binding to growth factor receptors. Expression of the proto oncogenes c-fos, c-jun, c-ha-ras and c-myc was studied in in vivo grown and in vitro cultured bovine preimplantation blastocysts employing RT-PCR, ribonuclease protection assay and immunohistochemistry. Thirteen- and 14- day-old preimplantation blastocysts, i.e. stages before and during trophoblast elongation, were used. In in vivo-grown blastocysts c-fos, c-jun and c-ha-ras transcripts as well as c-Fos, c-Jun and c-Myc proteins were detected in all stages studied. Cultured blastocysts were treated with 10 nM epidermal growth factor and 10 nM transforming growth factor-alpha simultaneously. Epidermal growth factor and transforming growth factor-alpha treatment induced c-fos mRNA and c-Myc protein expression. The induction of downstream targets of the epidermal growth factor receptor by epidermal growth factor and transforming growth factor-alpha indicates a functional epidermal growth factor signal transduction pathway in elongating bovine blastocysts. PMID- 10839632 TI - Advanced oviductal development, transport to the preferred implantation site, and attachment of the blastocyst in captive-bred, short-tailed fruit bats, Carollia perspicillata. AB - The final stages of embryonic development in the oviduct, transport of the embryo to the uterus, and the initial stages of implantation have been examined in captive-bred Carollia perspicillata at the light and electron microscopic levels. Development progressed to the expanded, zona pellucida-free, blastocyst stage in the oviduct. The abundance of microvilli on the exterior of the trophoblast varied with the degree of blastocyst expansion and cell shape, and may function in part as a membrane reservoir. Cells of the blastocyst also typically contained many lipid droplets and prominent areas of cytoplasm occupied by finely granular material (probably glycogen) instead of organelles. In most females, closure of the uterine lumen occurred prior to, or around the time of, transport of the blastocyst to the usual implantation site and appeared to play a role in preventing transport of the blastocyst too far distally in the uterus. This was associated with increased endometrial edema, particularly in the fundic region of the simplex uterus, and the extravasation of many erythrocytes into the endometrial stroma. Both of these changes began while the blastocyst was still being held in the oviduct and became pronounced during implantation. Engulfment of these erythrocytes by processes of the endometrial stromal cells and their phagocytosis by macrophages was also observed. Implantation was usually initiated within narrow tubular segments, lined by endometrium, that were located between the end of each oviduct and the main cavity of the uterus, or from immediately adjacent areas of the main cavity. During the early stages of implantation, the blastocyst was clasped by the endometrium at the implantation site, and this was associated with extensive interdigitation of the microvilli of the trophoblast and adjacent uterine epithelial cells. Initial adhesion of the trophoblast, which was still cellular rather than synctial, occurred over the apical intercellular junctions of the uterine epithelial cells. PMID- 10839633 TI - Arteether-induced brain injury in Macaca mulatta. I. The precerebellar nuclei: the lateral reticular nuclei, paramedian reticular nuclei, and perihypoglossal nuclei. AB - Malaria poses a threat across several continents: Eurasia (Asia and parts of Eastern Europe), Africa, Central and South America. Bradley (1991) estimates human exposure at 2,073,000,000 with infection rates at 270,000,000, illnesses at 110,000,000, and deaths at 1,000,000. Significant mortality rates are attributed to infection by the parasite Plasmodium falciparum, with an estimated 90% among African children. A worldwide effort is ongoing to chemically and pharmacologically characterize a class of artemisinin compounds that might be promising antimalarial drugs. The U.S. Army is studying the efficacy and toxicity of several artemisinin semi-synthetic compounds: arteether, artemether, artelinic acid, and artesunate. The World Health Organization and the U.S. Army selected arteether for drug development and possible use in the emergency therapy of acute, severe malaria. Male Rhesus monkeys (Macaca mulatta) were administered different daily doses of arteether, or the vehicle alone (sesame oil), for a period of either 14 days, or 7 days. Neuropathological lesions were found in 14 day arteether treated monkeys in the precerebellar nuclei of the medulla oblongata, namely: (1) the lateral reticular nuclei (subnuclei magnocellularis, parvicellularis, and subtrigeminalis), (2) the paramedian reticular nuclei (subnuclei accessorius, dorsalis, and ventralis), and the perihypoglossal nuclei (n. intercalatus of Staderini, n. of Roller, n. prepositus hypoglossi). The data demonstrate that the simina meduallry precerebellar nuclei have a high degree of vulnerability when arteether is given for 14 days at dose levels between 8mg/kg per day and 24 mg/kg per day. The neurological consequences of this treatment regimen could profoundly impair posture, gait, and autonomic regulation, while eye movement disorders might also be anticipated. PMID- 10839634 TI - Monoclonal antibody stains oligodendrocytes and Schwann cells in zebrafish (Danio rerio). AB - We prepared a monoclonal antibody that recognizes oligodendrocytes and Schwann cells in zebrafish. On immunoblots, the antibody mainly recognized three protein bands of 34 kDa in a membrane fraction from adult zebrafish brain. Medaka fish (Oryzias latipes) also possessed the same protein bands in a membrane fraction. The antibody did not stain neurons, but stained cells in fiber tracts and cranial and spinal nerves. In order to determine the nature of these cells, the staining pattern of the monoclonal antibody was compared with that of a myelin basic protein antiserum. Both antibodies stained oligodendrocytes and Schwann cells in fixed sections from the adult zebrafish. Both antigens were also co-localized in cultured glial cells. Taken together, these results indicate that the new monoclonal antibody recognizes myelinating glial cells in zebrafish and will be useful for the analysis of piscine glia. PMID- 10839635 TI - Parvalbumin immunoreactive Cajal-Retzius and non-Cajal-Retzius neurons in layer I of different cortical regions of human newborn. AB - Neurons of layer I play an important role in the development of the basic structural and functional organization of the mammalian cerebral cortex. Basic data, however, concerning the spatial and temporal distribution of the neuron populations in layer I are still limited, especially for human material. The present study investigates the distribution of Cajal-Retzius (CR) and non Cajal Retzius (NCR) neurons in thirteen cortical areas in the newborn human in terms of their relative density and possible subtypes. Neuronal populations were identified by immunohistochemistry for parvalbumin. Three main results are reported. First, parvalbumin-immunoreactive (Parv-ir) CR cells were observed in all of the neocortical areas examined. These areas also had a Parv-ir horizontal fiber plexus in deep layer I, confirming to the horizontal plexus classically associated with CR neurons. Second, many Parv-ir CR cells showed clear signs of degeneration. Third, in addition to the large CR cells, smaller Parv-ir NCR neurons occurred in many of the neocortical areas examined. These were morphologically heterogeneous and may represent several subtypes. By sampling across several areas, we were able to establish that these NCR cells occurred at higher density in primary sensory areas 3, 1, 17, and 41. Because of this variability in density of Parv-ir NCR cells, the ratio of Parv-ir CR to Parv-ir NCR cells is selectively lower in primary sensory areas. Recent investigations in somatosensory cortex of early postnatal rat report complex spatiotemporal patterns of correlated spontaneous activity among neurons in layer I (Schwartz et al. 1998). An interesting possibility is that regional variability in this activity may play a major role in the organization of cortical circuitry in different areas. PMID- 10839636 TI - A novel method for analysis of the periodicity of chondrogenic patterns in limb bud cell culture: correlation of in vitro pattern formation with theoretical models. AB - To experimentally examine whether the pattern changes predicted by theoretical models of pattern formation actually occur in a limb bud cell culture system, we developed a practical method to automatically measure the periodicity of chondrogenic patterns in vitro. The method utilizes binary image processing to quantify the total number of peak and valley pixels in a pattern to obtain the average interval between stripes in the chondrogenic pattern, and we named it the peak length method. The reliability of the peak length method was examined by using computer simulation results. The peak length method enabled us quantitatively obtain the average interval between chondrogenic islands, and the values obtained by this method were closely correlated with the average intervals obtained by manual measurement and two-dimensional Fourier transformation. The average intervals obtained by the peak length method were shown to be stable over a wide range of pattern variations that are frequently observed in actual experiments. By applying the peak length method to actual experimental data, we compared the validity of two theoretical models of pattern formation (cell sorting model and reaction-diffusion model) and it was concluded that the peak length method is a useful tool to quantitatively analyze chondrogenic patterns in limb micromass culture and to relate theoretical predictions and experimental results of pattern formation. PMID- 10839637 TI - The teleost kidney expresses Trk neurotrophin receptor-like proteins. AB - Neurotrophins are involved in the development and maintenance of some neuronal populations, acting through tyrosine protein kinase (Trk) receptors, TrkA, TrkB and TrkC. In addition, recent studies have demonstrated the occurrence of Trk receptors in a wide variety of adult and embryonic non-neuronal tissues in vertebrates, including kidney. Nevertheless no data are available on Trk proteins distribution in teleost kidney. The present study, by using an immunoperoxidase technique, analyses Trk receptor protein distribution in the kidney of three species of freshwater and saltwater teleost. TrkA-like immunoreactivity was the only one detected in all examined species, while TrkB-like immunoreactivity was completely absent and TrkC-like immunoreactivity was detected only in goldfish. The TrkA immunoreactive cells were mainly localised in the collecting duct system, as this system was the only one to also display TrkC. Such data could provide new clues to Trk in fish and aid assessment of the role of Trk protein receptors during vertebrate evolution. PMID- 10839638 TI - Does being female affect one's sleep? PMID- 10839639 TI - Bone marrow transplant for breast cancer is questioned on basis of incomplete data. PMID- 10839640 TI - Public education for women's health. PMID- 10839641 TI - Pregnancy Risk Assessment Monitoring System (PRAMS): unintended pregnancy among women having a live birth. PRAMS Working Group. PMID- 10839642 TI - Skin care--part I. PMID- 10839643 TI - Toward optimal health: the experts respond to skin cancer. Interview by Jodi Godfrey Meisler, M.S., R.D.. PMID- 10839644 TI - Nutrition--the best opportunity for improving the health of women. PMID- 10839645 TI - Treatment of depression during pregnancy. AB - Increasing numbers of patients are being treated for mood disorders. The majority of these patients, particularly with the diagnosis of major depression, are women of childbearing years. Concerns about fetal exposure to medication, both planned and unplanned, are becoming more pressing in the clinical practices of both psychiatrists and primary care physicians. There are relatively few study data available to guide clinicians in the use of psychotropic medications during pregnancy because of obvious problems in designing studies of the effects of medication on pregnant women, fetuses, and infants. Clinicians in all specialties receive little or no formal training in this area of psychopharmacology. This article gathers clinically relevant studies and practice information and provides suggestions regarding the approach to treatment of mood disorders during pregnancy, based on a risk assessment model. PMID- 10839646 TI - Osteoporosis and African American women. AB - The incidence of osteoporosis and related fractures in African American women is half that of Caucasian women. African American women who sustain osteoporosis related fractures have increased disability and decreased survival. Given the exponential increase in hip fracture rate among African American women over the age of 70 years, the risk of osteoporosis among this population may be underestimated. This review focuses on racial differences in women's bone mineral density (BMD) and bone metabolism and on various explanations for these observed differences. Environmental risk factors for osteoporosis and related fractures among African American women and modalities for prevention and treatment of osteoporosis are discussed. African American women begin menopause with higher BMD and have lower rates of women's bone loss after menopause, which account for their decreased incidence of osteoporosis and related fractures. The risk factors for osteoporosis among African American women are similar to those found in Caucasian women. Lifestyle interventions, such as calcium and vitamin D supplementation, smoking cessation, and increased physical activity, should be encouraged to enhance peak bone mass and to decrease bone loss. These interventions and other treatment modalities, such as hormone replacement therapy, bisphosphonates, and selective estrogen receptor modulators, should be studied further in African American women. PMID- 10839647 TI - Coronary risk factors in women one year after coronary artery bypass grafting. AB - Given the importance of risk factor management to long-term outcome following coronary artery bypass grafting (CABG) and the paucity of information on risk factor changes in women, a study was undertaken to examine the coronary risk factor status of women before and 1 year after CABG. This study was a prospective investigation of 130 women who underwent first-time, isolated CABG between February 1992 and October 1993. Lipid profiles, blood pressure, weight, smoking status, and other lifestyle behaviors were measured at the time of surgery and again 12 months later. The sample was 24% African American and had a mean age of 65 years and an average of 11 years of education. Substantial favorable changes in risk factor status occurred in the prevalence of smoking and the number of cigarettes smoked per day among smokers. Although the women experienced weight loss, 58% continued to be obese, and the self-reported dietary intake of fat, saturated fat, and cholesterol remained above the recommended levels of the National Cholesterol Education Program's Step II diet. Mean systolic and diastolic blood pressures significantly increased, and a substantial number of patients (54%) continued to exhibit hypertension at 1 year. No significant changes in plasma lipid concentrations were observed. At 1 year, one third of the women exceeded recommended levels for triglycerides, 78% for total cholesterol, and 92% for low-density lipoproteins. These findings indicate that women continue to have multiple coronary risk factors after CABG, putting them at high risk for future coronary heart disease events. Healthcare professionals need to target these women for effective secondary prevention. PMID- 10839648 TI - The influence of a low-fat diet on incidence and severity of migraine headaches. AB - Migraine headaches are a common, debilitating syndrome causing untold suffering and loss of productivity. A review of the literature indicates that high levels of blood lipids and high levels of free fatty acids are among the important factors involved in triggering migraine headaches. Under these conditions, platelet aggregability, which is associated with decreased serotonin and heightened prostaglandin levels, is increased. This leads to vasodilation, the immediate precursor of migraine headache. A high-fat diet is one factor that may directly affect this process. This study, undertaken to evaluate the impact of dietary fat intake on incidence and severity of migraine headache, was conducted over a 12-week period on 54 previously diagnosed migraine headache patients. During the first 28 days, the study subjects recorded all food consumption in a diet diary and maintained a headache diary. At the conclusion of this 28-day baseline period, subjects were individually counseled to limit fat intake to no more than 20 g/day. A 28-day run-in period was allowed for adaptation to the low fat diet. Results are reported on the final 28-day postintervention period. Subjects significantly decreased the ingestion of dietary fat in grams between baseline (mean 65.9 g/day, p < 0.0001) and the postintervention period (mean 27.8 g/day). The decreased dietary fat intervention was associated with statistically significant decreases in headache frequency, intensity, duration, and medication intake (all p < 0.0001). There was a significant positive correlation between baseline dietary fat intake and headache frequency (r = .44, p = 0.02). This study indicates that a low-fat diet can reduce headache frequency, intensity, and duration and medication intake. PMID- 10839649 TI - Sex differences in renal transplantation in Canada. AB - To determine if a patient's sex influences access to renal transplantation in Canada, transplant recipient data for first cadaveric unrelated renal transplants were obtained from the Canadian Organ Replacement Register (CORR) for the period 1985-1992. There were 4683 first unrelated cadaveric transplant recipients during this time. Differences in the proportion of men and women registered with CORR who received a renal transplant were analyzed. In Canada between 1985 and 1992, 25% of males 40 years and older on dialysis received renal transplants compared with 18% of females (p < 0.0001, RR 1.54, 95% CI 1.40-1.67). There was no difference in the rates of transplants in males and females who were under 40 years of age. Adjusting for panel-reactive antibody data did not change the significance of the difference in transplant rates between the sexes. In Canada from 1985 to 1992, male patients with end-stage renal disease received proportionately more transplants than females. PMID- 10839650 TI - A comparison of the effect of estrogen with or without progesterone on mood and physical symptoms in postmenopausal women. AB - Using a randomized, placebo-controlled design, this study assessed the effects of estrogen alone (ES) or in combination with cyclic progesterone (EP) on daily ratings of mood and physical symptoms before and after 6 months of daily hormone treatment. Fifty-four postmenopausal women were recruited from the community at large and specifically selected as being asymptomatic at the time of enrollment and without significant psychiatric history. Ratings were obtained every day for 30 days prior to treatment and again every day during the last 30 days of treatment. Results revealed that when compared with pretreatment levels, women randomized to EP (n = 16) exhibited statistically significant increases in daily depression, cramping, and breast tenderness and a marginally significant increase in daily anxiety. However, these increases were mild, not clinically significant, and did not interfere with normal functioning. Women randomized to ES (n = 20) showed no significant change in daily mood measures, although they did experience an increase in breast tenderness with estrogen. A significant placebo (n = 18) effect was observed, as there was a reliable reduction in daily irritability ratings (p < 0.05) with placebo. These findings suggest that for most postmenopausal women, the use of hormones will not be associated with clinically significant changes in mood or physical symptoms, which weighs favorably into the cost-benefit ratio for women considering hormone replacement therapy. PMID- 10839651 TI - Menstrual history and bone density in young women. AB - Adequate levels of reproductive and pituitary hormones are needed for the initiation and maintenance of regular menstrual cycles as well as for the achievement of peak bone mineral density (BMD). Therefore, in the absence of direct hormone measures, menstrual history may serve as a surrogate for the adequacy of hormonal functioning and be a marker for bone status in young women. In our cross-sectional study of white college women aged 19-26 years, we examined the association of six characteristics of menstrual history with bone density at the lumbar spine and the femoral neck. To characterize associations, we used multiple linear regression models that also accounted for the contribution of body mass index, dietary calcium intake, height, level of physical activity, smoking, and alcohol use. The associations between each of the six menstrual characteristics and BMD were stronger at the lumbar spine than at the femoral neck. Age at menarche explained the most variance at both the lumbar spine (partial r2 x 100 = 5.9%) and the femoral neck (partial r2 x 100 = 2.1%). For each year that menarche was delayed, bone density was lower by -0.023 g/cm2 (p = 0.0024) at the lumbar spine and -0.0129 g/cm2 (p = 0.0565) at the femoral neck. At the lumbar spine, a higher number of lifetime menstrual cycles was also significantly associated with increased bone density (adjusted beta = 0.0010, p = 0.0052, partial r2 x 100 = 4.4%). This association was not significant after adjusting for age at menarche. Neither reproductive years (age - age at menarche) nor a history of irregular cycles (either at menarche, in the past year, or ever) was associated with bone density at either site. Menstrual function appears to affect the bone density of these young women. Studies that include measures of reproductive and pituitary hormones are needed to further explore the role of hormones in the potential link between menstrual history and bone density. PMID- 10839652 TI - HIV and AIDS relative to other health, social, and relationship concerns among low-income urban women: a brief report. AB - Our objective was to determine the priorities of low-income women regarding health, relationship, and social concerns. Street-intercept surveys were conducted with 161 low-income urban women (mean = 27 years, 85% African American, 80% single mothers) regarding their perceptions of the threat associated with 48 health and social problems. Women rated acquired immune deficiency syndrome (AIDS) as their most important health, social, or relationship concern. Nearly one half of all women indicated that they would attend risk reduction programs to learn how to avoid infection with human immunodeficiency virus (HIV). AIDS is perceived as a serious threat to women's health, and interventions to reduce risk are welcomed. PMID- 10839653 TI - Is premenstrual dysphoric disorder a distinct clinical entity? AB - Does the evidence now available support the concept of premenstrual dysphoric disorder (PMDD) as a distinct clinical disorder such that the relative safety and efficacy of potential treatment can be evaluated? In a roundtable discussion of this question, a wealth of information was reviewed by a panel of experts. The key characteristics of PMDD, with clear onset and offset of symptoms closely linked to the menstrual cycle and the prominence of symptoms of anger, irritability, and internal tension, were contrasted with those of known mood and anxiety disorders. PMDD displays a distinct clinical picture that, in the absence of treatment, is remarkably stable from cycle to cycle and over time. Effective treatment of PMDD can be accomplished with serotinergic agents. At least 60% of patients respond to selective serotonin reuptake inhibitors (SSRIs). In comparison with other disorders, PMDD symptoms respond to low doses of SSRIs and to intermittent dosing. Normal functioning of the hypothalamic-pituitary-adrenal (HPA) axis, biologic characteristics generally related to the serotonin system, and a genetic component unrelated to major depression are further features of PMDD that separate it from other affective (mood) disorders. Based on this evidence, the consensus of the group was that PMDD is a distinct clinical entity. Potential treatments for this disorder can now be evaluated on this basis to meet the clear need for effective therapy. PMID- 10839654 TI - Women's Health Australia: on the progress of the main cohort studies. AB - More than 41,000 women aged 18-23, 45-50, and 70-75 years in 1996 are participating in the Australian Longitudinal Study on Women's Health (Women's Health Australia). Baseline surveys were conducted for all three cohorts in 1996, and the first follow-up survey of the mid-age group in 1998 has achieved a response rate exceeding 90%. The main findings so far reflect the large differences in the life experiences of the three cohorts. The young women report high levels of stress. The physical and mental health of those with young children is worse than for those without children, but confounding by sociodemographic characteristics may account for the differences. Two thirds of young women in the healthy weight or underweight range would like to weigh less, and early onset of dieting is associated with poorer physical and mental health. Most of the women in the mid-age group have multiple roles--in paid work, home duties, and caring for children and other dependents. The potential of the study to investigate the long-term impact of such busy lives on health outcomes is considerable. At this stage, the main health issues for these women relate to tiredness, weight gain, and menopause. The older cohort presents a picture of positive aging. These women are heavier users of health services than the mid-age and younger women, and they are also more satisfied with these services. Although their physical health is poorer, their mental health is better, and they report less stress than women in the other two cohorts. The follow-up survey of this group, planned for 1999, will focus on the coping strategies used by these women. An overall goal of the project is to understand the interactions among social roles, life events, and women's health in order to provide a basis for improved health policies and services. Analysis of these interactions, which relies on both quantitative and qualitative data, poses many challenges that will be addressed as the longitudinal data become available. PMID- 10839655 TI - Autoimmune diseases. PMID- 10839656 TI - Inhibition of murine AIDS by alternate administration of azidothymidine and fludarabine monophosphate. AB - Anti-HIV-1 combination therapies, including protease and reverse transcriptase inhibitors, can reduce plasma viremia to undetectable levels within the first 2 weeks of treatment. This reduction is followed by a slower decline that primarily results from the presence of viral reservoirs such as CD4+ memory cells, dendritic cells, and macrophages. For this reason, we evaluated a new drug combination therapy that includes a lympholytic drug: (2-fluoro-ara-AMP, fludarabine) to eliminate cells already infected and an antiviral drug (azidothymidine [AZT]) to protect cells not yet infected. We used C57BL/6 mice infected with the retroviral complex LP-BM5, which developed severe immunodeficiency (i.e., murine AIDS), to select the most effective fludarabine regimen to inhibit disease progression, and then to evaluate the efficacy and toxicity of the fludarabine and AZT combinations. The results obtained show that intraperitoneal administration of fludarabine at 3 mg/mouse twice a day for 4 weeks is the most effective regimen in reducing splenomegaly, lymphadenopathy, hypergammaglobulinemia, and proviral DNA content in spleen and lymph nodes and in restoring the architecture of lymph nodes. Subsequently, we evaluated the combined or sequential administration of fludarabine and AZT. The data reported in this paper show that the sequential administration of the two drugs provides additive antiviral effects that reduce spleen and lymph node weights to normal values and proviral DNA content by approximately 95% in all infected organs; the phenotypes of blood T and B cells moved toward control values, although the number of B cells was significantly reduced by fludarabine treatment. Finally, we evaluated the outcome of the disease after suspension or continuation of different treatment regimens. In all treatment groups, the disease progressed and increased proviral DNA content was found in infected organs, but animals receiving the sequential administration of fludarabine and AZT were less affected than those receiving only fludarabine or the simultaneous administration of both. The results obtained suggest that fludarabine could be part of a new therapeutic approach aiming at eradicating HIV from those cells that have been already infected and that are not protected by highly active antiretroviral therapy (HAART). PMID- 10839657 TI - Efavirenz- and adefovir dipivoxil-based salvage therapy in highly treatment experienced patients: clinical and genotypic predictors of virologic response. AB - OBJECTIVE: To determine the impact of prior nonnucleoside reverse transcriptase inhibitor (NNRTI) therapy, genotypic resistance, and other variables on response to efavirenz (EFV)- and adefovir dipivoxil (ADV)-based salvage therapy. DESIGN: Retrospective clinical cohort study. SETTING: One university and one community based HIV clinic. STUDY SUBJECTS: All 33 patients who were coenrolled in both the EFV and ADV expanded access programs. INTERVENTIONS: Patients received EFV 600 mg/day and ADV 120 mg/day in addition to other antiretroviral agents. OUTCOME MEASURE: HIV viral load (<500 copies/ml) at 12 and 24 weeks. RESULTS: 10 of 33 (30%) patients at 12 weeks and 8 of 33 (24%) patients at 24 weeks had viral loads <500 copies/ml. Prior NNRTI use and a history of any NNRTI-associated mutations predicted failure. Patients with Y181C or G190A single mutations had an initial greater magnitude of viral load suppression than those with K103N, but this advantage was short lived. No one with any NNRTI mutations responded with a viral load <500 copies/ml at 12 or 24 weeks. CONCLUSIONS: EFV/ADV-based salvage yielded viral load suppression at 24 weeks in 42% (8 of 19) of patients who were highly NRTI and protease inhibitor experienced but NNRTI naive. NNRTI-experienced study subjects had a poor response regardless of the specific NNRTI resistance mutation they harbored. PMID- 10839658 TI - Phase II placebo-controlled trial of fozivudine tidoxil for HIV infection: pharmacokinetics, tolerability, and efficacy. AB - Fozivudine tidoxil (FZD) is a thioether lipid-zidovudine (ZDV) conjugate with anti-HIV activity demonstrated in vitro and in pilot phase I studies. To assess its safety, efficacy and pharmacokinetics, we conducted a multicenter, randomized, double-blind, placebo-controlled trial of FZD monotherapy in 72 HIV infected patients who had not previously received antiretroviral therapy. In each dosage group (200 mg daily, 400 mg daily, 200 mg twice daily, 800 mg daily, 400 mg twice daily, and 600 mg twice daily), 12 patients were randomized to receive in a 10:2 ratio either FZD or a placebo for 4 weeks. Overall, FZD was well tolerated in all dosage groups; only 1 patient discontinued the drug, because of a moderate rise in aminotransaminase activity. HIV viral load fell in all the patients who were receiving FZD, except in the 200 mg daily group. The largest decrease (-0.67 log10) was observed in the 600 mg twice daily group. The plasma half-life was significantly longer (approximately 3.8 hours) than that of the parent drug ZDV. Exposure to ZDV, as reflected by the area under the time concentration curve, was much lower after FZD than after ZDV administration. FZD thus appears to be as effective as and potentially better tolerated than ZDV during short-term administration and has the advantage of once daily intake. PMID- 10839659 TI - Incidence of adverse reactions in HIV patients treated with protease inhibitors: a cohort study. Coordinamento Italiano Studio Allergia e Infezione da HIV (CISAI) Group. AB - OBJECTIVE: To assess the probability that protease inhibitor (PI) therapy might be discontinued because of adverse events (AE) and to evaluate the incidence rate of adverse reactions during PI treatment. DESIGN: A prospective cohort, multicenter study on HIV-positive patients starting treatment with at least one PI. SETTING: Ten departments of infectious diseases in Northern Italy. PATIENTS: A total of 1207 patients who started PI therapy in September 1997 and were consecutively observed up to April 1999. MAIN OUTCOME MEASURES: Adverse reactions following initiation of PI therapy, and time to therapy discontinuation due to AE. RESULTS: During the study period, 35.9% patients presented adverse reactions of any grade, whereas 9.7% presented at least one serious AE. After 12 months of treatment, the percentage of patients who had interrupted treatment was 36% of ritonavir-treated patients, 14.2% of those treated with indinavir, 13.6% of ritonavir-saquinavir hard gel capsules (HGC)-treated patients, and 8.5% and 2.1%, respectively, for those treated with nelfinavir and saquinavir HGC. Women and patients with hepatitis experienced a significantly greater number of adverse events compared with other categories. Gastrointestinal events were more frequently observed in patients treated with either ritonavir alone or in combination with saquinavir HGC, as well as in patients receiving nelfinavir, although in this group serious events were rare. Here again, neurologic, metabolic, and hepatic toxicity occurred more frequently in ritonavir and ritonavir-saquinavir HGC treated patients. Allergic reactions were more often observed in patients receiving nelfinavir. Indinavir-treated patients presented the highest incidence of renal toxicity. CONCLUSION: Ritonavir is the drug associated with the largest number of reactions, which appear during the first few months of treatment. Saquinavir HGC and nelfinavir are the best tolerated drugs in a clinical setting. PMID- 10839660 TI - Randomized trial of vitamin supplements in relation to vertical transmission of HIV-1 in Tanzania. AB - BACKGROUND: Observational studies suggest that poor nutritional status among HIV infected pregnant women is associated with a higher risk of vertical transmission of HIV. METHODS: We randomized 1083 pregnant women infected with HIV-1 in a double-blind, placebo-controlled trial to examine the effects of supplements of vitamin A and/or multivitamins (excluding vitamin A) using a 2-x-2 factorial design. We report the effects of the supplements on HIV infection defined using polymerase chain reaction (PCR), or death up to 6 weeks postpartum. RESULTS: Of babies in the multivitamin arm 38, (10.1%) were HIV-positive at birth compared with 24 (6.6%) in the no-multivitamin arm (relative risk [RR] = 1.54; 95% CI, 0.94-2.51; p = .08). Of babies born to mothers in the vitamin A arm, 38 (10.0%) were HIV-positive at birth compared with 24 (6.7%) in the no-vitamin A arm (RR, 1.49; 95% CI, 0.91-2.43; p = 0.11). Neither multivitamins nor vitamin A had an effect on HIV status at 6 weeks among those who were HIV-negative at birth (RR = 1.04; 95% CI, 0.65-1.66; p = 0.88) and (RR = 1.30; 95% CI, 0.80-2.09; p = .29, respectively). Similarly, neither supplement was associated with being either HIV infected or dead at birth (RR, 0.98; 95% CI, 0.76-1.27; p = .89 and RR, 1.01; 95% CI, 0.78-1.31; p = .95, respectively. A beneficial effect of multivitamins on birth weight was limited to babies who were HIV-negative at birth; babies in the multivitamin arm weighed +94 g more compared with those in the no-multivitamin arm (p = .02). Among babies who were HIV-positive at birth, the corresponding difference was -31 g (p = .82). CONCLUSIONS: Vitamin A and multivitamins did not affect the risk of vertical transmission of HIV in utero nor during the intrapartum and early breastfeeding periods. Multivitamins resulted in a significant improvement in birth weight of babies who were HIV-negative at birth but had no effect among those who were HIV-positive. The effect of vitamin supplements on HIV transmission through breastfeeding and on clinical progression of HIV disease is yet to be ascertained. PMID- 10839661 TI - Dual nucleoside regimens in nonadvanced HIV infection: prospective follow-up of 130 patients, Aquitaine Cohort, 1996 to 1998. Groupe d'Epidemiologie Clinique du SIDA en Aquitaine (GECSA). AB - OBJECTIVE: To describe the response to combinations of two nucleoside reverse transcriptase inhibitors (NRTIs) initiated early in the course of HIV infection under routine circumstances and to research prognostic factors indicating good virologic response. SETTING: Patients of the Aquitaine Cohort, a hospital-based open cohort that had been recruiting since 1987 in five public hospitals of the Aquitaine region in southwestern France. METHODS: Prospective cohort study of antiretroviral-naive patients with CD4+ cell counts >0.350 x 10(9)/L who started dual NRTI therapy between January 1996 and June 1997. Intent-to-treat analysis and multivariate logistic regression were used with data collected up to March 31, 1998. RESULTS: In this study, 130 patients were enrolled with a median follow up of 14 months. At the time of first prescription, 79% were in U. S. Centers for Disease Control and Prevention (CDC) group A, 16% in group B, and 5% in group C; median CD4+ cell count was 0.466 x 10(9)/L and median HIV RNA level was 4.52 log10 copies/ml. The two main combinations used were zidovudine (AZT) plus zalcitabine (ddC; 38%) and AZT plus didanosine (ddI; 37%). At week 52, median CD4+ and HIV RNA responses were, respectively, +80 cells and -1.6 log; the proportions of patients with HIV RNA level <5000 and <500 copies/ml were 70% and 45%, respectively, and 96% of the patients had a CD4+ cell count >0.350 x 10(9)/L at that time. At their last follow-up, 3 patients had reached been diagnosed with full-blown AIDS and the AIDS-free survival probability at 1 year was 98.2% (95% confidence interval [CI], 93.1-99.6); 1 death had occurred. The only significant variable associated with an undetectable HIV RNA level at 1 year was a lower HIV RNA level at the first prescription of dual therapy. CONCLUSION: Our data indicate that dual nucleoside combinations could be a therapeutic option for patients diagnosed and observed during follow-up in the early course of HIV infection. PMID- 10839662 TI - Serum triglycerides, HIV infection, and highly active antiretroviral therapy, Aquitaine Cohort, France, 1996 to 1998. Groupe d'Epidemiologie Clinique du Sida en Aquitaine (GECSA). AB - The aim of this study was to identify factors associated with serum triglyceride (TG) evolution in HIV-1-infected patients when highly active antiretroviral treatment (HAART) with or without protease inhibitors (PI) was introduced. Among 3191 patients of the Aquitaine Cohort (multirisk, both genders, multiple treatment patterns) observed during 1996 through 1998, 1429 had at least two measurements of TG, viral load, and CD4 cell count. Median follow-up was 21 months (interquartile range [IQR], 11-26) and median number of TG measures was 6 (IQR, 3-10). Median TG at baseline was 1.32 mmol/L (IQR, 0.91-2.05) and increased significantly over time (+2.5% for 100 days; 95% confidence interval [CI], 1.9 3.1). Longitudinal analysis of variations of TG was performed using mixed models. In crude analysis, baseline TG was higher in men, in those aged over 36 years, and in homosexuals. The following time-dependent variables were associated with an increase of TG: body weight increasing to >65 kg, diagnosis of AIDS, CD4 cell count falling to <50 cells/mm3, viral load falling to <500 cp/ml, and introduction of nucleoside analogues and PIs. In multivariate analysis, age >36 years (change of +17% of the TG level; 95% CI, 11-24), homosexuals (+13%; 95% CI, 4-23), AIDS stage (+12%; 95% CI, 5-19), weight >65 kg (+7%; 95% CI, 2-12) and PI (+21%; 95% CI, 17-27) remained significant. Factors identified before the availability of PI remain important but HAART with PI is a new major contributing factor to increased TG levels. PMID- 10839663 TI - Combination therapies for HIV and sexual risk behavior among gay men. AB - OBJECTIVE: To examine optimism in the light of recent advances in HIV treatment among gay men and its association with sexual risk behavior. METHODS: An anonymous questionnaire was completed by gay men who visited gyms in central London in March and April 1998 regarding their HIV status, unprotected anal intercourse (UAI) in the previous 3 months, and their response on a five-point linear scale to two measures of optimism: "I am less worried about HIV now that treatments have improved," and "I believe that new drug therapies make people with HIV less infectious." RESULTS: Two thirds of the men (67.5%, 522 of 773) did not agree with the statement, "I am less worried about HIV now that treatments have improved," and only 42 (5.4%) said they agreed quite a lot or a lot. HIV positive men were more likely to agree with this statement than HIV-negative men (p = .001) and men who had never been tested (p < .001). There was no association between agreement with this statement and frequency of UAI among HIV-positive or never-tested men (p > .3); there was, however, a positive association among HIV negative men who reported UAI with a partner of unknown or discordant status (p = .003). The vast majority of men (81.4%; 634 of 779) did not agree with the statement, "I believe that new drug therapies make people with HIV less infectious." Regardless of HIV status, no significant association was seen between agreement with this statement and frequency of UAI (p > .1 for all comparisons). CONCLUSION: Most gay men surveyed in central London gyms did not endorse the optimism statements concerning improved treatments or reduced infectivity. Although HIV-positive gay men were more likely to be optimistic than other men, there was no association between their optimism and sexual risk behavior. Among HIV-negative men, optimism around improved treatments (but not around reduced infectivity) was associated with UAI with a partner of unknown or discordant HIV status. For some HIV-negative men in London, optimism in the light of recent advances in HIV treatment may have triggered, or have been used as a justification for, sexual risk-taking. PMID- 10839664 TI - Correlates of risky behaviors among young and older men having sexual relations with men in Montreal, Quebec, Canada. Omega Study Group. AB - OBJECTIVE: To assess characteristics associated with 1) age and 2) recent unprotected anal sex with casual partners among men having sexual relations with men (MSM) participating in the Omega Cohort, Montreal, Quebec, Canada. METHODS: The Omega Cohort is a study of the incidence and psychosocial determinants of HIV infection among MSM living in Montreal. MSM complete a questionnaire and are tested for HIV every 6 months. RESULTS: Thirteen percent of young MSM (< 30 years of age, n = 355) and 12% of older MSM (> or = 30 years of age, n = 455) reported recent unprotected anal sex with casual partners. The predictors of this latter behavior were: not living with a male sexual partner, unprotected anal sex with regular partner, > 5 casual partners, alcohol/drug use before anal sex, and having difficulties with procedures needed for safe sex. Among young MSM, additional predictors were: to have been living in Montreal for less than 1 year and to have exchanged money for sex. Among older MSM, additional predictors were: female sexual partners, unprotected anal sex with an HIV-infected partner, and feeling invulnerable to AIDS. CONCLUSION: Young Omega participants do not have more risky behaviors than older participants. Some predictors of recent risk behaviors with casual partners were different between the two groups. Prevention programs should be adapted consequently. PMID- 10839665 TI - Emergence of zidovudine resistance in HIV-infected patients receiving stavudine. PMID- 10839666 TI - Evaluation of three rapid/simple tests for detection of HIV-2 antibodies. PMID- 10839667 TI - Persistent human herpesvirus 8 viremia associated with coinfection with human T cell lymphotropic virus type I and myelofibrosis. PMID- 10839668 TI - Development of a needs assessment system in rehabilitation. AB - The aim of this project was to develop a standardized, valid and reliable instrument for determining needs of patients and quality of care as perceived by patients. A questionnaire was constructed based upon the Service Quality Model. This model, patient expectations and experiences all play an important role in the questionnaire. The questionnaire contains a general part which includes ten determinants of care, the information the patient got and the perceived quality of care in general. The questionnaire also contains a specific part which includes questions about the actual care for diagnosis related problems. Twenty four spinal cord injured patients, staying at a Dutch rehabilitation centre, participated in this study. It is concluded that with the developed questionnaire the perceived quality of care according to the Service Quality Model can be assessed. Major contributions of the questionnaire to rehabilitation care will be its ability to highlight patient needs related to identified symptoms, and to provide a tool to signal aspects in the organization that can be improved. Thus it becomes a tool for quality management. Recommendations have been made for further research to make future use of the questionnaire in other settings and populations possible. PMID- 10839669 TI - Vocational rehabilitation--early versus delayed. The effect of early vocational rehabilitation compared to delayed vocational rehabilitation among employed and unemployed, long-term sick-listed people. AB - The aim of the present study was to examine whether the wait before vocational rehabilitation affected the outcome for employed and unemployed, long-term sick listed people. The study is based on sick-leave cases 90 days or longer, which started during 1992-1994 in a rural area in Sweden. The study included all sick leavers with non-malignant musculoskeletal diagnoses from the neck, shoulder and back regions and who had undergone vocational rehabilitation. Of these, 391 were employed and 78 unemployed. Our hypothesis was that those who undergo early vocational rehabilitation, irrespective of employment status, often get well sooner, have fewer sick days and lower benefit levels after vocational rehabilitation than those who have to wait a long time for their rehabilitation. Our hypothesis was supported only regarding the employed who mainly reported well at 6 and 12 months after vocational rehabilitation. In the longer term (24 months) the effect was however no longer evident. Earlier studies indicate that more factors than an early start to rehabilitation are important for a successful outcome. A more important factor is probably that the right measures have been initiated at the right time, that the measures are of high quality and that the sick-leaver is mentally prepared for the measure. Although early vocational rehabilitation did not prove to be a determining factor for a successful outcome there is still reason to emphasize early involvement in the case to guarantee rehabilitation of high quality. PMID- 10839670 TI - Changes in family functioning for stroke rehabilitation patients and their families. AB - This study charted changes over time in the family functioning of a group of stroke rehabilitation patients and their families. Participants were 60 12-month stroke survivors who had undergone an inpatient rehabilitation programme, 46 spouses, and 25 other family members. Participants were assessed at the time of admission to and discharge from rehabilitation, and 6 and 12 months after discharge. The study was undertaken in the rehabilitation unit at Repatriation General Hospital, in Adelaide, South Australia. Family functioning was assessed using the McMaster Family Assessment Device, and functional outcome was monitored with the competence scale of the Australian ADL Index. Longitudinal change in family functioning was observed in problem solving, family communication, role definition, behaviour control, and affective involvement. There were no significant differences between the family functioning ratings provided by patients and their spouses. It is important that consideration be given to means by which the increase in family dysfunction following stroke can be addressed. PMID- 10839672 TI - Rehabilitation outcome following traumatic brain injury--the Singapore experience. AB - The objectives of this study were to define (1) demographic and injury characteristics for acute traumatic brain injury (TBI) patients admitted to rehabilitation; (2) admission and discharge functional status; (3) discharge disposition and vocation status at follow-up. The charts of 80 consecutive TBI admissions (64 males, 16 females) were reviewed. Outcome measures included admission and discharge Modified Barthel Index (MBI) scores, length of rehabilitation stay, discharge disposition and return-to-work status at 1 year post-injury. Fifty-seven (71.3%) of patients were aged 40 years or less. Motor vehicle accidents accounted for 70.0% (56) of injuries. Sixty-one (76.3%) had severe head injuries. Diffuse cerebral oedema was present in 41.3% (33) and 52.5% (42) had intracranial haematoma. Admission and discharge MBI scores were 49.6 +/- 33.7 and 76.9 +/- 25.6 respectively. This difference was significant (P < 0.001). Memory and planning/organization deficits were present in 68.6% and 70.5% of patients (51 tested). Multiple regression analysis showed that admission MBI score had significant influence on discharge functional outcome (Multiple R 0.66, R2 0.40, P < 0.001). Seventy-two (90%) were discharge home and 25% (20) returned to work at 1 year post-injury. We conclude that this cohort of TBI patients showed significant improvement in functional status after rehabilitation, and admission functional status had the most impact on discharge functional outcome. PMID- 10839671 TI - Podiatric care for diabetic patients with foot problems: an observational study. AB - The aims of this study were to describe podiatric care for diabetic patients with foot problems and to explore the changes in knowledge, self-care behaviour and physical functioning after podiatric care. The treatment characteristics of 26 diabetic patients referred to podiatry were assessed. Prior to the first podiatric visit (T1) and 20 weeks later (T2) these patients filled in a structured questionnaire and performed a six-minute walking test. In half the number of patients preventive goals were set and strived for by general education about the diabetic foot and advice on footwear and self-care behaviour. With regard to treatment, reduction of pain was the most frequently selected goal. To achieve this reduction, a variety of interventions was applied. After podiatric care, patients reported having less severe foot pain and some improvements in functional ability and self-care behaviour were found. This study offers clues to start controlled clinical trials on the effectiveness of podiatry for diabetic patients. Trials should not only be directed to (the role of podiatry in) ulcer healing; it may be even more significant to study its effectiveness for the purpose of prevention and treatment of early-stage diabetic foot symptoms. PMID- 10839673 TI - Delinquency and social changes in United Arab Emirates: a comparison study between delinquent males and females on the family functioning questionnaire. AB - The purpose of this study was to specify descriptive elements of juvenile delinquents in United Arab Emirates (UAE); a comparison between male and female young offenders was also conducted using the Family Functioning Questionnaire (FFQ). The results revealed that some of the characteristics of young offenders were: school dropouts, had parents with no or some elementary education, lived in the city, and came from low income families. The statistical analysis revealed that there were significant differences between males and females on the FFQ subscales. That is, there was a differential treatment of male and female delinquents in the UAE society. Suggestions, implications and limitations for the potential use of the results reported in this study are also discussed. PMID- 10839674 TI - Treatment based on H-reflexes testing improves disability status in patients with cervical radiculopathy. AB - BACKGROUND: Postural modification in patients with lumbosacral radiculopathy either causes further H-reflex suppression, indicating increased root compression, or it effects recovery, indicating decompression of the spinal root. The posture that effects maximum recovery of the H-reflex amplitude is called optimum spinal posture (OSP) and is suggested as a therapeutic exercise to decompress the compromised nerve root. The focus of this study was to identify the OSP that effects the maximum recovery of the flexor carpi radialis (FCR) H reflex and to study its effect on the disability status in patients with cervical radiculopathy. SUBJECTS AND METHODS: Fourteen patients (46 +/- 12 y) with confirmed symptoms of C7 radiculopathy for the previous 6 months volunteered for the study. The FCR H-reflex was elicited by electrical stimulation of the median nerve at the cubital fossa (0.5 ms, 0.2 pps at H-max). Signals from the FCR muscle were recorded using a Cadwell 5200A EMG unit. The FCR H-reflex was recorded in natural sitting position with the head in natural position and in the OSP. Four traces of the H-reflex were recorded and averaged. The disability status was evaluated, using the Neck Disability Index (NDI), before exercising in the OSP and after 2 days of exercise in the OSP. DATA ANALYSIS: Paired t-test and Spearman's correlation coefficients were used. RESULTS: The H-reflex amplitude and latency were significantly different in the OSP and with the head in a natural position (P < 0.004; P < 0.011). Larger reflex amplitude and shorter latency were recorded in the OSP. The NDI scores were considerably improved after exercising in the OSP (P < 0.001). Spearman's correlation coefficient showed negative association between the H-reflex amplitude and the NDI scores (r = -0.64 to -0.54; P < 0.05). CONCLUSION: Exercising in the OSP increased the H-reflex amplitude and decreased latency of the compromised cervical root. It resulted in decreasing the disability status in this group of patients. PMID- 10839675 TI - The organization of vocational rehabilitation: the structure of success. AB - Focusing on vocational rehabilitation goals provides an understanding of the underlying structural and behavioural factors affecting successful employment outcomes. Two types of vocational centres having contrasting rehabilitation employment orientations are examined. Both types have similar end goals but organizational structures shaping divergent means toward the gainful employment of disabled persons. Utilizing a split sample of 14 vocational rehabilitation programmes in Israel, 7 sheltered and 7 supported programmes, a structured questionnaire of key administrative personnel and employees was generated. Success was based on six consensus measures. Clear differences between administrative, organizational and programme goals emerge, which affect goal priority and source of success measurement. Employing regression models, three keys to success are detected which stress the interplay between internal and external organization reference criteria in affecting success attainment. The results suggest that the organization of vocational rehabilitation has a direct impact on rehabilitation success. PMID- 10839677 TI - Well-being and disability: self-reports from students at a tertiary institution. PMID- 10839676 TI - New blind mobility aid devices based on the ultrasonic Doppler effect. AB - An original approach to electronic blind mobility aid combined with ultrasonic wave processing and spatial hearing has allowed the design of new blind mobility aid devices. These devices convert inaudible ultrasound echoes into audible sound via the Doppler effect, preserving even slight modification of the high frequency wave, so as to provide the user with higher acuity information regarding his movement, as well as object movement and their spatial location. PMID- 10839678 TI - Pedestrian skills training for children with learning difficulties. PMID- 10839679 TI - Recognizing child disability. PMID- 10839680 TI - Treatment of self-injurious behaviour by differential reinforcement and physical restraint. PMID- 10839681 TI - Better scholarship "on" teaching. PMID- 10839682 TI - Managed care education: a needs assessment of employers and educators of nurses. PMID- 10839683 TI - Jumping through hoops and walking on egg shells or discrimination, hazing, and abuse of students with disabilities? PMID- 10839684 TI - Influencing aspects in nursing education on Swedish nursing students' choices of first work area as graduated nurses. AB - It is difficult to recruit RNs to positions in various areas of elder care. The aim of this study is to understand the meaning of Swedish nursing students' reasoning during education about where in the health care system they would like to work as RNs after graduation. The students were interviewed using the same guide at the end of each of their three academic years. In the second and the third year the students kept diaries about their clinical education. A phenomenological-hermeneutic method of analysis was used, and eight themes appeared vital for the students' choice of work area after graduation. The deeper interpretations of the results imply that the students received contradictory messages during the education in elder care. Students found that nurses working in this field were often isolated with no apparent support system, which in turn reinforced their own ambivalence and reluctance towards future work in elder care. PMID- 10839685 TI - An evidence-based project for evaluating strategies to improve knowledge acquisition and critical-thinking performance in nursing students. AB - This longitudinal, quasi-experimental study with 142 junior nursing students focused on measurement of learning outcomes in two areas: acquisition of knowledge and development of critical thinking skills. The variation in clinical teaching strategy (structured versus unstructured health pattern assessment) was the independent variable. Results indicated significant gains in both knowledge and critical thinking performance from the beginning to the end of the semester. The significant gains in critical thinking performance provides support to the assertion that domain-specific measures of critical thinking are needed in nursing education. Additionally, our results suggested that it was the interaction between learning strategy and the characteristics of the learner that was more significant in determining knowledge improvement than the particular strategy. As a result of this study, it is recommended that faculty develop and use an evidence-based model to support their decision making regarding teaching methodologies. This seems especially relevant for large, introductory clinical courses that use team teaching to achieve educational goals related to improvement in critical thinking or knowledge. PMID- 10839686 TI - Preparing nursing students for contemporary practice: restructuring the psychomotor skills laboratory. AB - The restructured laboratory experience offered a safe environment that supported student experimentation with psychomotor skills and self-initiated approaches to problem solving. Restructuring psychomotor laboratory experiences with emphasis on communication and conceptualization of principles supported students to begin addressing clinical problems with flexibility, creativity, and the premise for lifelong skill acquisition. Students who have skills that extend beyond technique will inevitably be better prepared to meet the demands of health care systems and patients now and in the future. PMID- 10839687 TI - The skills of coach are an essential element in clinical learning. AB - Students learn about the nursing profession through experience and close association with expert practitioners. It has been found repeatedly that expert nurses as clinical teachers "coach" students to realize their full potential. The steps of coaching have been described here to provide guidelines for clinicians new to the teaching role. The activity of coaching in clinical education is worthy of further investigation. Not only do students develop confidence in psychomotor skills, coaching appears to facilitate scholarship in the clinical setting. Skilled coaches encourage students to form meaningful, coherent theoretical frameworks for the information gathered. Without the support of a coach, students would continue to adhere to familiar views of the world and accept the clinical world as it presents itself without healthy scepticism or the ability to be creative. Cognitive coaching is not a separate activity to psychomotor coaching. Together both strategies offer a unique, and to date a poorly described, method to communicate practice knowledge to the nursing student. The ultimate benefit of coaching is the ability to instill in the student the motivation to improve performance and strive for excellence. PMID- 10839688 TI - Predicting academic success of transfer nursing students. PMID- 10839689 TI - Comparing videotaped and faculty-present return demonstrations of clinical skills. PMID- 10839690 TI - Redressing the future. PMID- 10839691 TI - Caution, balance needed in recommending herbals over hormone replacement. PMID- 10839692 TI - Sexual health in mature women. PMID- 10839693 TI - Nurses' study disproves link of fat to breast cancer. PMID- 10839694 TI - The women's health research coalition: a new advocacy network. AB - The Society for the Advancement of Women's Health Research is a nonprofit, nonpartisan organization committed to improving the health of women through research. The Society was founded in 1990 to bring national attention to the problem of the exclusion of women from clinical trials and the need for greater federal funding for diseases and conditions prevalent in and unique to women. Today, the Society works closely with members of the public and private sectors and academic institutions to promote a women's health research agenda. The Society seeks both to shape public policy and improve public and professional education. The Journal of Women's Health is the official publication of the Society. We welcome your views. Contact us by fax at (202) 833-3472, or visit our website at http://www.womens-health.org/. PMID- 10839695 TI - von Willebrand Disease in women: the need for recognition and understanding. PMID- 10839696 TI - Patient education: allergy care. PMID- 10839697 TI - Toward optimal health: the experts respond to allergies. Interviews conducted by Jodi Godfrey Meisler. PMID- 10839698 TI - In pursuit of a healthy diet: directions and detours. PMID- 10839699 TI - IPPF Charter on Sexual and Reproductive Rights. International Planned Parenthood Federation. AB - For most of human existence and in most societies, women have been considered to be property and subject to men. Throughout history, with such notable exceptions as Queen Boadicea, Eleanor of Aquitaine, Elizabeth I of England, and Catherine the Great of Russia, women had little or no power until early in the 20th century when the women's suffrage movement was successful in the United States and in some European countries. As women have gained political rights, groups of women have sought sexual and reproductive rights, as exemplified by the feminist movement of the past few decades in the United States. Although marked strides toward achievement of reproductive choice have been taken in high-income countries, there remain major strictures to reproductive freedom for women in low income countries. This area, which is replete with ethical and moral issues, has been addressed by the International Planned Parenthood Foundation (IPPF), which has worked to improve the sexual and reproductive health of women throughout the world. The IPPF Charter on Sexual and Reproductive Rights is a paradigm for both women's rights and human rights. Karen Newman is policy adviser with the IPPF and has codrafted the IPPF Charter on Sexual and Reproductive Rights together with two lawyers. She has held several positions within the IPPF, including medical researcher, press officer, and programme adviser in Europe, where she had responsibility for working with new family planning associations (FPAs) in the Czech Republic and Slovakia. At present, she is working to increase the capacity of IPPF member FPAs to undertake human rights-based advocacy for sexual and reproductive health and rights. Judith F. Helzner is director of Sexual and Reproductive Health at International Planned Parenthood Federation/Western Hemisphere Region, Inc., where she has worked since 1987. She holds M.A. degrees from the University of Pennsylvania in International Relations and Demography. Her previous employment includes the Pathfinder Fund and the International Women's Health Coalition. PMID- 10839700 TI - Should women have a different risk assessment from men for primary prevention of coronary heart disease? PMID- 10839701 TI - Papanicolaou smear: can we make a good test better? Technical and interpretive challenges for the practitioner. AB - The Papanicolaou smear has proven to be an excellent screening test for cervical cancer and has resulted in a significant decrease in mortality from invasive cervical cancer. A relatively high false-negative rate of Pap smears might cause failure to diagnose preinvasive disease, especially when coupled with an increased screening interval. This review focuses on the causes of and methods to reduce the number of false-negative Pap smears. In addition, we address the clinical significance of ambiguous Pap smear diagnoses, such as atypical squamous/glandular cells of undetermined significance, and suggest a scheme for management of patients with these diagnoses. Although efforts to reduce false negative Pap smears are important, the most effective way of reducing mortality from invasive cervical cancer is universal, organized, population-based screening of all women. PMID- 10839702 TI - The psychological ramifications of weight management. AB - It has long been believed that food restriction leads to psychological disturbances, including depression, preoccupation with food, and binge eating. However, recent studies suggest that comprehensive weight loss programs that incorporate behavioral treatment, diet change, and encouragement of physical activity in fact can improve the psychological state, including mood. A study conducted on subjects participating in the Weight Watchers program demonstrated positive psychological changes and improved quality of life. These changes may help motivate overweight people to maintain the physical activity and nutritional practices necessary to lose and maintain weight. Programs that include group support, like Weight Watchers, have been associated with psychological benefits independent of the amount of weight lost. Furthermore, dieters who regain lost weight do not appear to experience adverse psychological consequences. The development or exacerbation of bulimia has been linked by some authors to strict dieting, but more moderate weight control programs do not appear to produce disordered eating and may help reduce binge eating among overweight people. Individuals who successfully lose and maintain weight have been shown to experience improved mood, self-confidence, and quality of life. Additionally, decreasing levels of psychological and behavioral symptoms have been associated with increasing duration of weight loss maintenance. It can be concluded that quality of life and other psychological measures improve in individuals on comprehensive weight management programs. PMID- 10839703 TI - The treatment of obesity: what's new, what's recommended. AB - The prevalence of obesity in the United States has increased, with approximately one third of American men and women considered obese. Obese individuals who seek weight reduction frequently lose weight with the goal of returning to their ideal body weight. For the majority of obese persons, however, reaching their ideal body weight is an unattainable goal, few people are able to maintain even more modest weight losses over the long term. The result is that many obese persons end up feeling frustrated, if not defeated, by their weight loss efforts. Recent weight loss treatment recommendations have focused on helping patients lose 5% 10% of their body weight and maintaining these losses for longer durations. These more modest losses frequently are associated with reductions in obesity-related comorbidities and improvements in psychological status. The practitioner assisting obese individuals with weight reduction is presented with a variety of treatment options, including behavioral programs, pharmacotherapy, and low calorie diets. Each of these approaches has been shown to be effective in producing the 5%-10% weight losses now recommended. A combination of these approaches, such as the use of pharmacotherapy with a behavioral modification program, ultimately may prove to be the most effective treatment for obese persons. We review the new weight loss recommendations and provide suggestions for assessing patients for weight loss treatment. We also discuss both existing and new treatment options and focus on their application in a primary care setting. PMID- 10839704 TI - Eclampsia: a 13-year experience at a United States tertiary care center. AB - Eclampsia, the occurrence of a grand mal seizure in the setting of hypertension in pregnancy, remains a major women's health issue and an important cause of maternal and fetal morbidity in the United States. We reviewed the incidence, management, and outcome of cases of eclampsia during a 13-year period at a major maternity hospital. We confirmed 33 cases of eclampsia seen during that period and have evaluated risk factors in this population. Medical records were reviewed to obtain demographic and clinical data. Characteristics of the eclamptic women were compared with those of the general obstetric population during the same time period. The overall incidence of eclampsia at this tertiary care center was 0.028%. The majority of eclamptic women (75%) had four or more prenatal visits. Young age (< or = 20 years) and first pregnancy remained important risk factors for eclampsia. Although many women with eclampsia had preceding hypertension or elevated urine protein levels or both, some experienced eclampsia as their first disease manifestation. Although the occurrence of eclampsia was low, eclampsia continues to complicate pregnancy in this large U.S. obstetric population. PMID- 10839705 TI - The association of body fat distribution with lifestyle and reproductive factors in women aged 45-54 in Kinmen County, Republic of China. AB - This community-based cross-sectional study was conducted in Kinmen County, R.O.C. The purpose of this study was to investigate the factors that influence Chinese women's body fat distribution, including demographic, lifestyle, reproductive, and anthropometric variables. Participants were 1310 women aged 45-54 years. We measured waist/hip ratio (WHR), age, education level, occupation, alcohol consumption, diet, physical exercise, smoking, age at menarche, parity, menopausal status, age at menopause, and obesity (body mass index, BMI). Lower BMI, premenopausal status, and higher education level were primary correlated factors of gynecoid pattern body fat distribution. Higher BMI and postmenopausal status were the main correlated factors of android pattern body fat distribution. Lifestyle factors did not show any significant association with body fat distribution. As in Western countries, overall obesity and menopausal status are the determinants of body fat distribution. Controlling overall obesity is the key factor that can be addressed in public education programs to reduce the risk of high WHR, which plays an important role in the development of some chronic diseases. PMID- 10839706 TI - Confusion about mammography: prevalence and consequences. AB - Over the last decade, there has been significant controversy about the schedule on which women, particularly women in their 40s, should have mammograms. The purpose of the analysis reported here was to assess whether women in their 40s and 50s were confused as a result of the controversy following the January 1997 National Institutes of Health Consensus Development Conference on Breast Cancer Screening For Women Ages 40-49. We also examined if confusion was related to being off schedule for mammography. The study sample included 1287 women recruited from a random sample of 2165 Blue Cross/Blue Shield of North Carolina members. The data described in this analysis were derived from a baseline telephone interview conducted as part of a larger intervention trial. Study measures included a variety of sociodemographic, medical, belief, and behavioral variables. Overall, 28% of women were confused, and 35% were off schedule. Although a higher proportion of women in their 40s than 50s were confused, more women in their 50s were off schedule. Confusion was a significant predictor for the outcome being off schedule. Predictors of confusion included several belief variables, risk perceptions, age (40s), whether the woman had a regular physician, and whether she had enough information about mammography. Healthcare providers should ask some simple questions to determine if women are confused and then seek to meet their information needs. PMID- 10839707 TI - Decisions about prenatal testing for chromosomal disorders: perceptions of a diverse group of pregnant women. AB - We conducted a study to elucidate factors influencing women's decisions regarding prenatal genetic screening for and diagnosis of chromosomal disorders and to learn about their experiences with these tests and with the medical system. Using focus group interviews and questionnaire assessments, we obtained detailed impressions of a diverse group of 75 pregnant women. Participants varied with respect to race/ethnicity, religious background, and reproductive history, as well as in their decisions about use of prenatal screening and diagnostic testing. Substantial variation surfaced in attitudes toward testing. Factors influencing women's views included available resources, feelings about having a child with Down syndrome, moral beliefs, family and social influences, perceptions of one's own health, the difficulty of becoming pregnant, and willingness to put the fetus at elevated miscarriage risk. Such findings indicate that age-based policies regarding access to prenatal diagnoses that, among other reasons, are based on the balance of risks between bearing a child with a chromosomal abnormality versus procedure-related loss are incompatible with the range of concerns that women bring to this decision and the weight individual women may assign to the outcomes. PMID- 10839708 TI - Differences in menstrual bleeding characteristics, functional status, and attitudes toward menstruation in three groups of women. AB - This report examines differences in bleeding characteristics, functional status, and attitudes toward menstruation among three groups of women: (1) women who complain of abnormal uterine bleeding (AUB), (2) women who have similar menstrual patterns as those complaining of AUB but who do not perceive themselves to have abnormal bleeding, and (3) women without evidence of prolonged or excessive bleeding. Women who complain of AUB and women with heavy bleeding but not complaining of AUB, although similar on two important menstrual symptoms (very heavy bleeding or episodes of unusually heavy bleeding) differ on a number of other menstrual characteristics, including the frequency of short cycles, the probability of having an abnormally long period, and reporting of unusually heavy bleeding lasting longer than 1 day. Whether women reported concerns with menstruation or not, the majority of women in this analysis had fairly negative attitudes toward menstruation. However, this negativity toward menstruation did not translate into women wanting a hysterectomy, even for those with heavy bleeding. The major difference among the three groups of women was the strong negative effect of AUB on functional status. A majority of women complaining of AUB reported that the bleeding interfered significantly with their daily routine, making them unable to function at work and at home. These results suggest that although the main complaint of women with AUB is very heavy bleeding, a number of other specific menstrual characteristics differentiate women with AUB from other women with very heavy bleeding who do not perceive the bleeding to be problematic. The complaint of AUB appears to be related to how significantly bleeding affects daily functioning. Therefore, an important factor to assess when considering treatment of AUB is the extent to which bleeding symptoms significantly affect functional status. PMID- 10839709 TI - Use of counseling by abused pregnant Hispanic women. AB - The purpose of this study was to determine if there are characteristics of abused women that are associated with the women's use of the services of counseling to help end the abuse. The research design was a prospective, descriptive analysis of 216 abused pregnant Hispanic women receiving prenatal care in urban public health clinics. Women who were abused by their intimate male partner were offered unlimited access to the services of a bilingual English/Spanish-speaking counselor experienced in abuse whose office was located in the public health clinic. Baseline demographic characteristics, severity of abuse experienced by the women, and the women's previous use of community resources were assessed. Analyses were conducted to examine associations between baseline characteristics and the number of visits the abused women made to the counselor. The results of the study show that when an abused pregnant Hispanic woman had an average of two or more children, she was more likely to use the services of an abuse prevention counselor. The study also showed that the women who had used the police most during the previous 12 months made the fewest number of visits to the counselor. Prenatal care may provide a convenient and safe setting for low-income, ethnic minority women with numerous children to conveniently and safely access abuse prevention services. Research is needed on the effectiveness of incorporating abuse intervention services into nontraditional settings, such as neighborhood clinics. PMID- 10839710 TI - Outcomes and health status of socially disadvantaged women during pregnancy. AB - We sought to examine the health status of disadvantaged pregnant women more broadly and to consider if poor maternal health predisposes a woman to an adverse birth outcome. We surveyed 250 women recruited from six health centers in the greater Boston area during their third trimester. The main predictor variables were maternal physical functioning (PF), emotional health (EH), and overall health status in the month prior to pregnancy. The main outcome variables were the decline of maternal PF and EH during pregnancy and adverse birth outcomes. Mean PF scores fell from 91.9 prior to pregnancy to 63.7 during the third trimester (mean scores transformed 0 to 100, where a higher score represents better health). EH remained unchanged during pregnancy. After adjustment, women with a preexisting medical condition reported a lower PF score prior to pregnancy (87.8 versus 94.5, p < 0.05). Poor PF prior to pregnancy or during the third trimester was associated with an increased risk of preterm labor (odds ratio 2.02, 95% confidence interval 1.03-3.97). This study is the first to employ general health status measures to examine changes in health during pregnancy. Our findings support the use of preconception care to improve the health status of disadvantaged women with pre-existing conditions. This study suggests that poor maternal health may predispose a woman to an increased risk of preterm labor. PMID- 10839712 TI - Women's health literaturewatch. PMID- 10839711 TI - Cholesterol. PMID- 10839713 TI - The 500 Dalton rule for the skin penetration of chemical compounds and drugs. AB - Human skin has unique properties of which functioning as a physicochemical barrier is one of the most apparent. The human integument is able to resist the penetration of many molecules. However, especially smaller molecules can surpass transcutaneously. They are able to go by the corneal layer, which is thought to form the main deterrent. We argue that the molecular weight (MW) of a compound must be under 500 Dalton to allow skin absorption. Larger molecules cannot pass the corneal layer. Arguments for this "500 Dalton rule" are; 1) virtually all common contact allergens are under 500 Dalton, larger molecules are not known as contact sensitizers. They cannot penetrate and thus cannot act as allergens in man; 2) the most commonly used pharmacological agents applied in topical dermatotherapy are all under 500 Dalton; 3) all known topical drugs used in transdermal drug-delivery systems are under 500 Dalton. In addition, clinical experience with topical agents such as cyclosporine, tacrolimus and ascomycins gives further arguments for the reality of the 500 Dalton rule. For pharmaceutical development purposes, it seems logical to restrict the development of new innovative compounds to a MW of under 500 Dalton, when topical dermatological therapy or percutaneous systemic therapy or vaccination is the objective. PMID- 10839714 TI - Novel keratin 16 mutations and protein expression studies in pachyonychia congenita type 1 and focal palmoplantar keratoderma. AB - Pachyonychia congenita type 1 (PC-1) is an autosomal dominant ectodermal dysplasia characterized by nail dystrophy, focal non-epidermolytic palmoplantar keratoderma (FNEPPK) and oral lesions. We have previously shown that mutations in keratin 16 (K16) cause fragility of specific epithelia resulting in phenotypes of PC-1 or FNEPPK alone. Here, we report 2 novel mutations in K16 causing distinct phenotypes. A heterozygous missense mutation (L124R) was detected in a kindred with PC-1. In a family where mild FNEPPK was the only phenotype, a 23 bp deletion and a separate 1 bp deletion downstream were found in exon 6: [1244-1266del; 1270delG]. At the protein level, these mutations remove 8 residues and substitute 2 residues in the helix termination motif (HTM) of the K16 polypeptide. The HTM sequence is conserved in all known intermediate filament proteins and for convenience, this complex mutation was designated deltaHTM. Transient expression of K16 cDNAs carrying either the L124R or the deltaHTM mutation in epithelial cell line PtK2 produced aggregation of the keratin cytoskeleton. However, the aggregates observed with the deltaHTM mutation were morphologically different and appeared to be less disruptive to the endogenous cytoskeleton. Therefore, loss of the HTM sequence may render this mutant K16 less capable of contributing to filament assembly and decrease its dominant-negative effect, resulting in the milder FNEPPK phenotype. PMID- 10839715 TI - Alternative subcellular locations of keratinocyte basonuclin. AB - Basonuclin is a zinc finger protein present in the basal cell layer of the epidermis and in hair follicles. Human basal epidermal cells are often heterogeneous with respect to a nuclear or cytoplasmic location of basonuclin and the protein may be concentrated in either compartment. In mouse and rat epidermis, although clusters of basonuclin may be seen in some basal cell nuclei, the protein is mainly concentrated in the cytoplasm. When epidermis whose basal cells contain predominantly cytoplasmic basonuclin is disaggregated and the cells are cultivated in the presence of supporting 3T3 cells, the basonuclin of the growing keratinocyte colonies is strongly concentrated in the cell nuclei. Transfer of the cells to culture medium without supporting 3T3 cells results in a predominantly cytoplasmic concentration of the basonuclin. This translocation is reversible, since addition of supporting 3T3 cells restores most basonuclin to the nucleus. The nuclear location is associated with more rapid cell growth. We conclude that different states of the keratinocyte require greater or less activity of basonuclin, and the subcellular location of the protein is probably related to the magnitude of its action on the cells. PMID- 10839716 TI - Quantitative analysis of UVB-induced apoptosis in human epidermis. AB - The quantitative measurement of the induction of apoptosis in cells grown in vitro can be accomplished using a variety of proven methods. However, the quantitative assay of apoptosis within an intact tissue is very laborious and the results can be misleading. We have established a method to quantitatively analyze the induction of apoptosis in human epidermis following UVB irradiation. The assay is based on the activation of the apoptotically induced enzyme caspase 3, using a synthetic caspase 3 substrate. The activation of caspase 3 was shown to correlate with the induction of apoptosis in human keratinocytes cultures as a monolayer. We then demonstrated that the activation of caspase 3 could be measured from UVB-irradiated whole skin. The induction of apoptosis was confirmed by cellular morphology and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling. Therefore, we concluded that the measurement of caspase 3 specific activity in UVB-irradiated human epidermis was an efficient, inexpensive, and accurate method to quantitate UVB-induced apoptosis in vivo. PMID- 10839717 TI - Cross-talk between epidermal growth factor receptor and protein kinase C during calcium-induced differentiation of keratinocytes. AB - The induction of epidermal differentiation by extracellular Ca2+ involves activation of both tyrosine kinase and protein kinase C (PKC) signaling cascades. To determine if the differentiation-dependent activation of tyrosine kinase signaling can influence the PKC pathway, we examined the tyrosine phosphorylation status of PKC isoforms in primary mouse keratinocytes stimulated to terminally differentiate with Ca2+. Elevation of extracellular Ca2+ induced tyrosine phosphorylation of PKC-delta, but not the other keratinocyte PKC isoforms (alpha, epsilon, eta, zeta). We have previously demonstrated that activation of the epidermal growth factor receptor (EGFR) pathway induces PKC-delta tyrosine phosphorylation in basal keratinocytes (Denning M F, Dlugosz A A, Threadgill D W, Magnuson T, Yuspa S H (1996) J Biol Chem 271: 5325-5331). When basal keratinocytes were stimulated to differentiate by Ca2+, the level of cell associated transforming growth factor-alpha (TGF-alpha) increased 30-fold, while no increase in secreted TGF-alpha was detected. Furthermore, Ca2+-induced tyrosine phosphorylation of PKC-delta and phosphotyrosine-association of the receptor adapter protein Shc was diminished in EGFR -/- keratinocytes, suggesting that EGFR activation may occur during keratinocyte differentiation. Tyrosine phosphorylated PKC-delta was also detected in mouse epidermis, suggesting that this differentiation-associated signaling pathway is physiological. These results establish a requirement for the EGFR in Ca2+-induced tyrosine phosphorylation of PKC-delta, and document the production of cell-associated TGF-alpha in differentiated keratinocytes which may function independent of its usual mitogenic effects. PMID- 10839718 TI - Calcium release activated calcium entry in a human skin derived cell line (HaCaT). AB - Using isolated cells from a spontaneously immortalized human keratinocyte cell line (HaCaT) loaded with Fura-2 the regulation of intracellular Ca2+ concentration ([Ca2+]i) was studied. The continuous presence of ATP induced a biphasic response in high external Ca2+. The first component reflected the release of calcium from intracellular stores since it was present after the removal of external calcium with ethylene-glycol-bis-N,N,N',N'-tetraacetic acid (EGTA). The second phase of [Ca2+]i increase was not detectable in the absence of external calcium and raising the extracellular [Ca2+] increased the rate of rise in [Ca2+]i suggesting that it was influenced by the external environment. Furthermore, after adenosine triphosphate (ATP) had emptied the intracellular store in a calcium-free milieu, the elevation of external Ca2+ induced a secondary increase in [Ca2+]i that did not require the presence of ATP. Depleting the intracellular calcium store with a Ca-ATP-ase inhibitor (cyclopiasonic acid, 10 microM) also induced calcium entry. The depletion induced calcium entry was inhibited by econazole (100 microM). These findings indicate the presence of a calcium release activated calcium influx pathway in HaCaT keratinocytes. PMID- 10839719 TI - Lanceolate hair-J (lahJ): a mouse model for human hair disorders. AB - Lanceolate hair-J (lahJ) arose spontaneously in 1994 on the DBA/1LacJ inbred background at The Jackson Laboratory. Mutant mice were runted, alopecic, and lacked vibrissae. As they aged, their skin wrinkled. Affected mice developed a noninflammatory, proliferative skin disease with follicular dystrophy. Hair fibers developed a number of abnormalities including periodic nodules along the shaft (trichorrhexis nodosa), compaction resembling trichorrhexis invaginata, spiral fractures, broken tips, and lance-shaped tips. This mutation exhibits some characteristics that resemble an autosomal recessive ichthyosiform disease that occurs in humans characterized in part by peculiar, invaginating, multinodal, hair shaft abnormalities known as Netherton's syndrome. Periodic nodules also resemble the human genetic based disease monilethrix. This autosomal recessive mouse mutation, allelic with lanceolate hair (lah), based on breeding studies, is located on mouse Chromosome 18, within a cluster of genes coding for adhesion molecules. Homozygotes for either of these allelic mouse mutations have elevated serum IgE levels, a feature also common with human Netherton's syndrome. PMID- 10839720 TI - Electrophysiological characterization of human keratinocytes using the patch clamp technique. AB - The living, not-cornified part of the epidermis consists mainly of keratinocytes. The control mechanisms of proliferation and differentiation are only partly understood. Similarly, the influence of ionic channels of the cell membrane on the proliferation of keratinocytes remains unclear. Preliminary investigations point to a relation between the opening of ionic channels and keratinocyte proliferation. Therefore, voltage clamp experiments were performed to gain further knowledge of the electrophysiological characteristics of human keratinocyte cellular membranes. In-vitro cultured keratinocytes of the cell line HaCaT were characterized by means of the voltage-clamp technique. As measured in the whole-cell configuration, changing the extracellular K+ or Cl--concentration shifted the membrane potential of HaCaT cells. Application of the patch-clamp technique in the cell-attached and inside-out configuration revealed an ionic channel with a conductance of multiples of 200 pS. The reversal potential of the single channel current was shifted by substituting of intracellular Cl- by aspartate-. Channel openings disappeared after addition of 0.1 mM of the anion channel blocker 4',4' diisothiocyanato-stilbene-2-2' disulfonic acid (DIDS). It is concluded that this channel contributes to the Cl--conductance of the cellular membrane and is a determinant of the membrane potential of human keratinocytes. This channel may represent a target for pharmacological manipulation of the membrane potential and possibly the growth of human keratinocytes in dermatological proliferation disorders. PMID- 10839721 TI - Identification of desmoglein 1 as autoantigen in a patient with intraepidermal neutrophilic IgA dermatosis type of IgA pemphigus. AB - In a 51-year-old female patient with intraepidermal neutrophilic IgA dermatosis (IEN) type of IgA pemphigus, circulating IgA, but not IgG, autoantibodies were detected to bind to the cell surface of the whole epidermis, being much stronger in the upper epidermis. In the patient's skin a heavy intraepidermal IgA staining was observed throughout the whole epidermis, accompanied by a weak IgG and a more prominent C3 staining. IgA from the patient's serum showed no reactivity either with epidermal proteins by immunoblot analysis, or with COS 7 cells transiently transfected with mammalian cell expression constructs containing full length human Dsc1, Dsc2 and Dsc3. Our patient's IgA specifically reacted with conformational epitopes of human desmoglein (Dsg) 1 but not Dsg 3, when studied in a previously established, here for IgA antibody detection modified enzyme linked immunoabsorbent assay (ELISA) of baculovirus expression system. The immunoreactivity against keratinocyte cell surface was completely removed from the serum of the patient by pre-incubation with recombinant Dsg1 baculoprotein. This finding indicates that the sera possess only IgA antibodies against the extracellular domain of Dsg1 baculoprotein, but no antibodies against components of keratinocyte cell surface other than Dsg1. This is the first case of IgA pemphigus where Dsg1 has been identified as the autoantigen. PMID- 10839722 TI - Preface. PMID- 10839723 TI - Chlamydia pneumoniae and cardiovascular disease: an evolutionary perspective on infectious causation and antibiotic treatment. AB - Evolutionary considerations implicate infectious causation of atherosclerosis and help to resolve different risk factors as parts of an overall process of disease causation. An evolutionary approach also provides insight for the timing of research efforts to provide better control of pathogen evolution. In particular, evolutionary considerations emphasize the need to understand the transmissibility of Chlamydia pneumoniae from systemic infections in order to control the evolution of antibiotic resistance. PMID- 10839724 TI - Background and current knowledge of Chlamydia pneumoniae and atherosclerosis. AB - Attributes of Chlamydia pneumoniae of potential importance to a relationship with atherosclerosis are described. Among these are that C. pneumoniae is not new. It is unique. It is a pathogen with which everyone is infected, and it is difficult to treat. It causes immunopathology, myocarditis, and endocarditis and chronicity is a hallmark of Chlamydia infection. Current knowledge of the relation of C. pneumoniae and atherosclerosis comes from observational (e.g., seroepidemiology and tissue studies) and experimental studies. The limitations of the serologic studies of chronic infection are noted as is the conclusive demonstration of an association of C. pneumoniae and atherosclerosis by the repeated and frequent finding of the organism in atherosclerotic tissue. Experimental studies are needed to determine if the association is causal. Such studies should include animal models, basic mechanisms, and secondary prevention antibiotic treatment trials. PMID- 10839725 TI - Epidemiologic association of Chlamydia pneumoniae and atherosclerosis: the initial serologic observation and more. AB - About 70% of persons with acute myocardial infarction (AMI) show a seroresponse to a chlamydial lipopolysaccharide (LPS) epitope. Elevated titers against Chlamydia pneumoniae in sera from such patients point to an exacerbation in a chronic infection as does a change in the nature of immune complexes containing chlamydial LPS. The presence of antibodies to C. pneumoniae proteins in immune complexes suggests an intimate association of the pathogen with the vascular system. In the first prospective study, elevated antibody titers or immune complexes containing chlamydial LPS were an independent significant risk factor (odds ratio, /=29 days were 1.0 (reference), 0.93 (0.81-1.07), 0.99 (0.81-1.20), and 1.03 (0.84-1.26), respectively. These results suggest little or no association between past use of erythromycin or tetracycline antibiotics and the risk of first MI among this population. PMID- 10839760 TI - Inflammation, infection, or both in atherosclerosis: the ROXIS trial in perspective. AB - The role of infection in the instability of atherosclerotic plaques has been questioned because of discrepancies in the results of clinical trials that tested antibiotics in acute coronary syndromes. The results of the Randomized Trial of Roxithromycin in Non-Q-Wave Coronary Syndromes (ROXIS) are summarized and contrasted with two other pilot studies of antibiotic therapy of coronary artery disease. Relevant characteristics of patients enrolled and rationales for these trials are discussed and serologic results are presented. PMID- 10839761 TI - The ACADEMIC study in perspective (Azithromycin in coronary artery disease: elimination of myocardial infection with Chlamydia). AB - Chlamydia pneumoniae, a common cause of respiratory infection, is vasotropic and frequently found in human atheromas. Whether it plays a causal role in coronary artery disease (CAD) is uncertain. The effects of 3 months of azithromycin treatment or placebo were tested in 302 patients with chronic CAD seropositive to C. pneumoniae at 3-6 months. Azithromycin reduced a global rank sum score of 4 inflammatory markers (C-reactive protein [CRP], interleukin [IL]-1, IL-6, tumor necrosis factor-alpha; P=.011) and a global rank sum change score (+/-SD) (from 535+/-201 to 587+/-190; P=.027) at 6 (but not 3) months. Change scores for CRP and IL-6 and median IL-1 levels were lower. C. pneumoniae IgG and IgA antibody titers were unchanged. Clinical cardiovascular events at 6 months did not differ between groups (azithromycin, 9; placebo, 7). Infections were reduced and drug was well tolerated. Thus, azithromycin caused modest but significant reductions in markers of inflammation, but differences in clinical events were not evident at 6 months. However, power was limited and conclusions should await results of the 2-year evaluation and larger studies. PMID- 10839762 TI - Rationale and design of a secondary prevention trial of antibiotic use in patients after myocardial infarction: the WIZARD (weekly intervention with zithromax [azithromycin] for atherosclerosis and its related disorders) trial. AB - Mounting evidence supports the contention that atherosclerosis is an inflammatory disease. Recently a possible role for infectious microorganisms has gathered attention. Chlamydia pneumoniae is one possible pathogen. If C. pneumoniae is a target organism, antibiotics with antichlamydial activity may be able to ameliorate plaque instability. The WIZARD trial is a secondary prevention study that is assessing the impact of a 3-month course of azithromycin compared with placebo on the progression of clinical coronary heart disease. The study will enroll 3300 patients who have had a prior myocardial infarction and who have a C. pneumoniae IgG titer of >/=1:16. The primary end point is a composite of time to either recurrent myocardial infarction, death, a revascularization procedure, or hospitalization for angina. This study is the first of a series of adequately powered clinical trials that will attempt to bridge insights from preclinical investigations to interventions applicable to patient care. PMID- 10839763 TI - Description and status of the azithromycin and coronary events study (ACES). AB - The Azithromycin and Coronary Events Study is a randomized, double-blind, placebo controlled trial of azithromycin among adults with stable coronary artery disease. The study is based on the hypothesis that infection with Chlamydia pneumoniae may be causally associated with cardiovascular disease and therefore that treatment directed against this organism may reduce the risk of subsequent coronary events. Participants randomized to treatment will receive 600 mg of azithromycin orally once a week for 1 year and will be followed a mean of 4 years for the composite primary outcome of coronary heart disease death, nonfatal myocardial infarction, hospitalization for unstable angina, and coronary revascularization. Secondary objectives include those related to a better understanding of the relationship between antibody titer and inflammatory markers with treatment status and outcome; therefore, all participants will have blood specimens obtained at enrollment and a random 25% will have additional specimens collected periodically during follow-up. PMID- 10839764 TI - Collaborative multidisciplinary workshop report: clinical antimicrobial trials for primary and secondary prevention of atherosclerotic cardiovascular disease. AB - The task assigned to the working group on Clinical Antimicrobial Trials for Primary and/or Secondary Prevention of Atherosclerotic Cardiovascular Disease was to evaluate the need for additional clinical antibiotic trials of a primary or secondary nature for the treatment of atherosclerotic heart disease and to suggest possible designs for future trials. In addition, the working group was to define the role of collaboration in answering research questions. PMID- 10839765 TI - What is needed to prove that Chlamydia pneumoniae does, or does not, play an etiologic role in atherosclerosis? PMID- 10839766 TI - How much "UFC" is really cortisol? PMID- 10839767 TI - Protein glycosylation and diseases: blood and urinary oligosaccharides as markers for diagnosis and therapeutic monitoring. AB - BACKGROUND: N- and O-oligosaccharide variants on glycoproteins (glycoforms) can lead to alterations in protein activity or function that may manifest themselves as overt disease. APPROACH: This review summarizes those diseases that are known to be the result of an inherited or acquired glycoprotein oligosaccharide structural alteration and that are diagnosed in blood or urine by chemical characterization of that oligosaccharide alteration. CONTENT: The biochemical synthesis steps and catabolic pathways important in determining glycoprotein function are outlined with emphasis on alterations that lead to modified function. Clinical and biochemical aspects of the diagnosis are described for inherited diseases such as I-cell disease, congenital disorders of glycosylation, leukocyte adhesion deficiency type II, hereditary erythroblastic multinuclearity with a positive acidified serum test, and Wiskott-Aldrich syndrome. We also review the laboratory use of measurements of glycoforms related to acquired diseases such as alcoholism and cancer. CONCLUSIONS: Identification of glycoprotein glycoforms is becoming an increasingly important laboratory contribution to the diagnosis and management of human diseases as more diseases are found to result from glycan structural alterations. PMID- 10839768 TI - Quantitative Beutler test for newborn mass screening of galactosemia using a fluorometric microplate reader. AB - BACKGROUND: The Beutler enzyme spot test is an effective assay for newborn mass screening of galactosemia, but it is qualitative and relies on visual interpretation. We describe a quantitative, instrumental modification of the assay. METHODS: We modified the macroscopic visual Beutler enzyme spot test by adding extraction of blood components from filter paper, deproteinization with acetone-methanol, and quantification and recording by a fluorescent microplate reader and personal computer. All handling was performed in microplates. The measurement time was 90 min. RESULTS: Fluorescence intensity (FI) of healthy controls correlated with hematocrit and galactose-1-phosphate uridyltransferase (GALT) activity. Patients with GALT deficiency were distinguished clearly from healthy subjects and heterozygous carriers by FI. FI decreased to 75% of the initial activity after storage at 25 degrees C for 3 days and to 40% after storage at 37 degrees C for 7 days. Screening of 46 742 newborns yielded 1 false positive result (in a heterozygous carrier), 1 patient with glucose-6-phosphate dehydrogenase deficiency, and no apparent false negatives as judged by concurrent measurements of galactose and galactose-1-phosphate. CONCLUSIONS: The quantitative Beutler test can provide precise GALT activity in newborn mass screening, and can take into consideration the influence of high temperature and humidity, duration between sampling and testing, and anemia. This method is clinically useful, simple, automated, and highly reliable for newborn mass screening of galactosemia. PMID- 10839769 TI - Detection of the Finnish-type congenital nephrotic syndrome by restriction fragment length polymorphism and dual-color oligonucleotide ligation assays. AB - BACKGROUND: Congenital nephrotic syndrome of Finnish type (NPHS1) is an autosomal recessive disorder characterized by severe proteinuria of intrauterine onset. Ninety-four percent of the Finnish NPHS1 chromosomes have been reported to carry either a 2-bp deletion in exon 2 (Fin(Major)) or a nonsense mutation in exon 26 (Fin(Minor)) of the NPHS1 gene. The high prevalence of only two mutations in the Finnish population enables the use of molecular techniques in the diagnosis of NPHS1 and for carrier screening. METHODS AND RESULTS: We describe two different molecular methods for the detection of the NPHS1 mutations: a PCR-restriction fragment length polymorphism (PCR-RFLP) and a dual-color oligonucleotide ligation assay (OLA). The dual-color OLA, which enables simultaneous detection of the NPHS1 Fin(Major) and Fin(Minor) mutations, can be used for rapid analysis of large sets of samples. The analysis of 2004 Finnish blood samples revealed 34 carriers of the Fin(Major) mutation and 1 carrier of the Fin(Minor) mutation, indicating a carrier frequency of 1:59 (95% confidence interval, 1:89-1:44) for the NPHS1 Fin(Major) mutation and 1:2004 (95% confidence interval, 0 to 1:677) for the NPHS1 Fin(Minor) mutation, respectively. CONCLUSION: PCR-RFLP and dual color OLA are suitable for molecular diagnosis and carrier screening of the major mutations that cause NPHS1. PMID- 10839770 TI - Troponin T and I assays show decreased concentrations in heparin plasma compared with serum: lower recoveries in early than in late phases of myocardial injury. AB - BACKGROUND: Heparinized plasma samples allow more rapid analysis than serum samples, but preliminary studies showed lower cardiac troponin T (cTnT) results in plasma. We undertook a multicenter study to characterize this effect for cTnT and cardiac troponin I (cTnI). METHODS: Blood samples were collected with and without heparin at five hospitals. cTnT was measured by a "third generation" assay (Elecsys((R))), and cTnI was measured by a commercial immunoassay (IMMULITE((R))). RESULTS: Mean cTnT was 15% lower in heparin sampling tubes than in serum. Measured concentrations of cardiac troponins also decreased with increasing heparin concentrations added to sera. Heparin-induced losses were greater in early than in late phases after onset of chest pain. Addition of heparin ( approximately 100 IU/mL) to serial samples from nine acute myocardial infarction patients produced mean cTnT losses of 33% at 1-12 h after onset of chest pain, 17% at 13-48 h, and 7% after 48 h. The changing heparin effects were seen for both cTnT and cTnI during time courses of individual patients with myocardial infarction. CONCLUSION: We suggest that binding of heparin to troponins decreases immunoreactivity, especially in early phases of myocardial injury. The resulting losses may depend on the antibodies used in each troponin assay. PMID- 10839771 TI - Age-related increases in plasma phosphatidylcholine hydroperoxide concentrations in control subjects and patients with hyperlipidemia. AB - BACKGROUND: The basal lipid peroxide concentration in the plasma of patients with hyperlipidemia may be related to atherosclerosis. Quantitative determination of lipid peroxides in the plasma is an important step in the overall evaluation of the biochemical processes leading to oxidative injury. Unfortunately, the currently available methods for lipid peroxidation lack specificity and sensitivity. METHODS: Hyperlipidemic patients (44 males and 50 females), ages 12 82 years (mean +/- SE, 53 +/- 2.3 years for males, 58 +/- 2.0 years for females, and 56 +/- 14 years for total cases), and normolipidemic volunteers (controls, 32 males and 15 females), ages 13-90 years (49 +/- 4 years for males, 65 +/- 4 years for females, and 55 +/- 24 years for total cases), were recruited in the present study. Plasma phosphatidylcholine hydroperoxide (PCOOH) was determined by chemiluminescence-HPLC (CL-HPLC). RESULTS: Plasma PCOOH concentrations increased with age in both controls and hyperlipidemic patients. However, the mean plasma PCOOH concentration in patients with hyperlipidemia (331 +/- 19 nmol/L; n = 94) was significantly (P <0.001) higher than in the controls (160 +/- 65 nmol/L; n = 47). Plasma PCOOH concentrations were similar in three hyperlipidemic phenotypes: hypercholesterolemia (IIa), hypertriglyceridemia (IV), and combined hyperlipidemia (IIb). The mean plasma PCOOH in patients with treatment-induced normalized plasma lipids was 202 +/- 17 nmol/L. There was no significant correlation between plasma PCOOH concentration and total cholesterol, triglycerides, or phospholipids in hyperlipidemic patients. For all subjects, there was a significantly positive correlation between plasma PCOOH and each lipid (total cholesterol, P = 0.0002; triglycerides, P = 0.0137; and phospholipids, P <0.0001). Analysis of fatty acids composition of plasma phosphatidylcholine showed significantly low concentrations of n-6 fatty acids moieties (linoleic acid and arachidonic acid) in patients compared with controls. CONCLUSIONS: Our results suggest that an increase in plasma PCOOH in patients with hyperlipidemia may be related to the development and progression of atherosclerosis, particularly in the elderly. Measurement of plasma PCOOH is useful for in vivo evaluation of oxidative stress. PMID- 10839772 TI - HPLC analysis of lipid-derived polyunsaturated fatty acid peroxidation products in oxidatively modified human plasma. AB - BACKGROUND: Lipid peroxidation is a prominent manifestation of free radical activity and oxidative stress in biological systems. Diverse methodologies have been developed that measure a variety of lipid peroxidation products used as markers of lipid peroxidation processes. METHODS: Hydroxy and hydroperoxy polyunsaturated fatty acid (PUFA) peroxidation products were analyzed in human blood plasma by reversed-phase HPLC after liquid-liquid extraction of total lipids and alkaline hydrolysis of lipid esters to liberate free PUFAs. An isocratic mobile phase containing 1 g/L acetic acid-acetonitrile-tetrahydrofuran (52:30:18, by volume) over 60 min duration, with ultraviolet absorbance detection at 236 nm by photodiode array, enabled the resolution and quantification of 13 regioisomeric hydroxy and hydroperoxy PUFAs. RESULTS: As little as 250 microL of human plasma was utilized with an analytical range of 0.033-1.6 micromol/L for each compound. Intra- and interassay CVs for all compounds detected in normal or oxidatively modified human plasma were 3.2-11% and 4.7-12%, respectively. Analytical recoveries were 87-103%. Analysis of human plasma exposed to artificial oxidation with Cu(2+) ion and hydrogen peroxide, a free radical generating reaction, showed marked increases in hydroxy and hydroperoxy PUFA concentrations. CONCLUSION: Lipid-derived hydroxy and hydroperoxy PUFAs may be useful as clinical markers of lipid peroxidation and oxidative stress in the peripheral circulation. PMID- 10839773 TI - Determination of gentamicins C(1), C(1a), and C(2) in plasma and urine by HPLC. AB - BACKGROUND: Gentamicin is an aminoglycoside antibiotic complex containing gentamicins C(1), C(1a), and C(2). Few methods have been described for analysis of the three gentamicin components separately in biological fluids, and none has been used in pharmacokinetic studies. Determination of the three gentamicins separately may have pharmacokinetic and toxicological implications. The present study describes development of an HPLC method for the analysis of gentamicin C(1), C(1a), and C(2) components in plasma and urine. METHODS: The three components were isolated by preparative chromatography and their identities verified by thin-layer chromatography, HPLC, mass spectrometry, nuclear magnetic resonance spectroscopy, and melting point determination. The gentamicins were extracted from the biological matrix by use of Tris buffer and polymer phase solid-phase extraction. Derivatization was carried out in the solid-phase extraction cartridge with 1-fluoro-2, 4-dinitrobenzene. The 2,4-dinitrophenyl derivatives were separated with reversed-phase HPLC and quantified by the ultraviolet absorbance at 365 nm. RESULTS: The detector response was linear from the limit of quantification to 50 mg/L for the individual components. The limit of quantification was 0.07 mg/L for gentamicin C(1) and 0. 1 mg/L for gentamicins C(2) and C(1a). The recovery of the gentamicin components was 72% from plasma and 98% from urine. The method was validated for human and dog plasma and urine. CONCLUSIONS: The method was repeatable and enabled the analysis of gentamicins C(1), C(1a), and C(2) in plasma and urine in concentrations covering the therapeutic range of the drug, thus being suitable for therapeutic drug monitoring and pharmacokinetic studies. PMID- 10839774 TI - Circulating immunoreactive proANP(1-30) and proANP(31-67) in sedentary subjects and athletes. AB - BACKGROUND: Atrial natriuretic peptide (ANP) is synthesized and stored in myocytes as prohormone(1-126), which upon release is cleaved into proANP(1-98) and alpha-ANP(99-126). In addition, cleavage of proANP(1-98) produces proANP(1 30), proANP(31-67), and proANP(79-98) fragments. ProANP(1-30) and proANP(31-67) have roles in fluid and electrolyte homeostasis. The aim of the present study was to develop a plasma assay for proANP(1-30) and proANP(31-67) and to compare results in trained athletes and sedentary subjects. METHODS: Two competitive enzyme immunoassays were established with affinity-purified sheep antiserum against synthetic ANP fragments. The immunoreactivity (ir) of proANP(1-30) and proANP(31-67) was measured in 10-microL plasma samples without extraction in a microwell-based assay. Plasma concentrations in sedentary male subjects (n = 22) and male endurance athletes (n = 14) were examined. RESULTS: In the assay for ir proANP(1-30) and ir-proANP(31-67), the concentrations at 95% B/B(0) were 4.7 and 14.2 pmol/L, respectively. Within-run CVs were 4-6% and 5-6%, and between-run CVs were 9% for both assays. Both assays were linear on dilution (y = 0.9945x - 0. 7291 and y = 1.0001x - 3.428), and the recoveries were 102-112% and 102-106%, respectively. In the sedentary and athletic groups, the ir-proANP(1-30) concentrations were similar: 318 +/- 38 pmol/L and 312 +/- 25 pmol/L (mean +/- SE), respectively, whereas the ir-proANP(31-67) was higher in the rowers (713 +/- 81 pmol/L) than in the sedentary subjects (387 +/- 71 pmol/L; P <0.005). CONCLUSIONS: The proANP fragment assays are precise (CV <10%) and exhibit nearly quantitative recovery (102-112%). Only ir-proANP(31-67) responds to physical training. PMID- 10839775 TI - Branched-chain keto-acids and pyruvate in blood: measurement by HPLC with fluorimetric detection and changes in older subjects. AB - BACKGROUND: Measurement of keto-acids is important in various clinical situations. The aim of the present work was to develop a rapid HPLC method for the determination of keto-acids in human serum and to assess the concentrations of these acids in young adults and institutionalized elderly adults. This method was applied to the determination of blood keto-acid concentrations of young adults and institutionalized elderly people, divided into age groups METHODS: Four keto-acids (alpha-ketoisocaproate, alpha-ketoisovalerate, alpha-keto-beta methylvalerate, and pyruvate) were derivatized with o-phenylenediamine to give fluorescent derivatives. After the sample preparation step (75 min to prepare 20 samples), the derivatives were separated chromatographically on a reversed-phase column using a binary gradient. RESULTS: The fluorometric detection of the four keto-acids was rapid, <12 min. The method is repeatable and reproducible: the CVs were <6% and <11%, respectively, for each of the keto-acids. We found no significant difference between males and females. Concentrations of the branched chain keto-acids decreased after age 60 years, especially alpha-ketoisocaproate, which decreased approximately 40%. CONCLUSIONS: The proposed method allows rapid and reliable measurement of keto-acids. The data demonstrate that changes in branched-chain keto-acids concentrations in serum occur with age. PMID- 10839776 TI - Leptin in cerebrospinal fluid from children: correlation with plasma leptin, sexual dimorphism, and lack of protein binding. AB - BACKGROUND: Previous studies in adults have established that leptin is present at very low concentrations in cerebrospinal fluid (CSF), but few data exist concerning CSF leptin in children. Current evidence suggests that CSF leptin concentrations interact with hypothalamic centers controlling food intake. Serum leptin concentrations manifest a sexual dimorphism that arises during puberty. METHODS: Leptin concentrations were determined in CSF from 42 pre- and postpubertal children who had been objectively classified into non-neurological disease or aseptic meningitis groups. Multivariate analysis of the dependence of CSF leptin on gender, pubertal state, body mass index (BMI), presence of aseptic meningitis, and CSF protein concentration was performed. RESULTS: CSF leptin concentrations correlated with log-transformed plasma leptin concentrations in concomitantly collected samples (r = 0.582; P = 0.029). BMI and gender were significant determinants of CSF leptin in postpubertal children, but only BMI was significant in prepubertal children. Analysis with HPLC to separate protein-bound and free forms of leptin found only free leptin in CSF. CONCLUSIONS: CSF leptin concentrations in children reflect plasma leptin concentrations, including the advent of sexual dimorphism at puberty. Only free leptin is detectable in CSF, suggesting that it is the biologically active form. PMID- 10839777 TI - Serum and urinary prostate-specific antigen and urinary human glandular kallikrein concentrations are significantly increased after testosterone administration in female-to-male transsexuals. AB - BACKGROUND: The genes that encode prostate-specific antigen (PSA) and human glandular kallikrein (hK2) are up-regulated by androgens and progestins in cultured cells, but no published studies have described the effect of androgen administration in women on serum and urinary PSA or hK2. METHODS: We measured serum and urinary PSA and hK2 before, and 4 and 12 months post testosterone treatment by immunofluorometric methods in 32 female-to-male transsexuals. RESULTS: Mean serum PSA increased from 1.1 ng/L to 11.1 ng/L and then to 22 ng/L by 4 and 12 months post treatment, respectively; the corresponding mean values in urine were 17, 1420, and 18 130 ng/L, respectively. Serum hK2, another kallikrein closely related to PSA, remained undetectable at the three time points. However, urinary hK2 concentration rose from below the detection limit (<6 ng/L) before treatment to 18 and 179 ng/L by the 4th and the 12th month of treatment, respectively. All changes were statistically significant (P <0.001) at 4 months. CONCLUSIONS: Testosterone administration increases serum and urinary PSA and urinary hK2 in women. These measurements may be useful as indicators of androgenic stimulation in women. PMID- 10839778 TI - Serum concentrations of intact parathyroid hormone in healthy children. PMID- 10839780 TI - Nonparametric estimation of reference intervals by simple and bootstrap-based procedures. PMID- 10839779 TI - Effects of hemoglobin C and S traits on seven glycohemoglobin methods. PMID- 10839781 TI - Estimation of serum apolipoprotein B by a modified homogeneous assay for HDL cholesterol. PMID- 10839782 TI - Latex-enhanced immunoturbidimetry allows D-dimer determination in plasma and serum samples. PMID- 10839783 TI - New approaches to cyclosporine monitoring raise further concerns about analytical techniques. PMID- 10839784 TI - Interference by glycolic acid in the Beckman synchron method for lactate: a useful clue for unsuspected ethylene glycol intoxication. PMID- 10839785 TI - Sensitive ELISA for human immunoglobulin D measurement in neonate, infant, and adult sera. PMID- 10839786 TI - Urinary free cortisol is unrelated to physiological changes in urine volume in healthy women. PMID- 10839787 TI - Rapid detection of angiotensinogen M/T235 polymorphism by fluorescence probe melting curves. PMID- 10839788 TI - Erythrocyte sedimentation rate by the test-1 analyzer. PMID- 10839789 TI - Improved sensitivity of capillary electrophoresis for detection of bisalbuminemia. PMID- 10839790 TI - Silver staining of denaturing gradient gel electrophoresis gels. PMID- 10839791 TI - Macroprolactin reactivities in prolactin assays: an issue for clinical laboratories and equipment manufacturers. PMID- 10839792 TI - Plasma ferritin in acute hepatocellular damage. PMID- 10839793 TI - A high factor II/Factor X functional ratio is not a useful predictor of the FII G20210A gene mutation in thromboembolic patients undergoing oral anticoagulant treatment. PMID- 10839794 TI - Genistein: a potent natural antiandrogen. PMID- 10839798 TI - Revising B cell receptors. PMID- 10839799 TI - Assessment of caspase activities in intact apoptotic thymocytes using cell permeable fluorogenic caspase substrates. AB - To detect caspase activities in intact apoptotic cells at the single cell level, cell-permeable fluorogenic caspase substrates were synthesized incorporating the optimal peptide recognition motifs for caspases 1, 3/7, 6, 8, and 9. Caspase activities were then assessed at various times after in vitro treatment of mouse thymocytes with dexamethasone or anti-Fas antibody. Dexamethasone induced the following order of appearance of caspase activities as judged by flow cytometry: LEHDase, WEHDase, VEIDase, IETDase, and DEVDase. Since the relative order of caspases 3 (DEVDase) and 6 (VEIDase) in the cascade has been controversial, this caspase activation order was reexamined using confocal microscopy. The VEIDase activity appeared before DEVDase in every apoptotic cell treated with dexamethasone. In contrast, anti-Fas stimulation altered this sequence: IETDase was the first measurable caspase activity and DEVDase preceded VEIDase. In an attempt to determine the intracellular target of the potent antiapoptotic agent carbobenzoxy-valyl-alanyl-aspartyl(beta-methyl ester)-fluoromethyl ketone (Z VAD[OMe]-FMK), we examined its ability to inhibit previously activated intracellular caspases. However, no significant reductions of these activities were observed. These fluorogenic caspase substrates allow direct observation of the caspase cascade in intact apoptotic cells, showing that the order of downstream caspase activation is dependent on the apoptotic stimulus. PMID- 10839800 TI - Signaling via beta(2) integrins triggers neutrophil-dependent alteration in endothelial barrier function. AB - Activation of polymorphonuclear leukocytes (PMNs) and adhesion to the endothelial lining is a major cause of edema formation. Although known to be dependent on the function of beta(2) integrins (CD11/CD18), the precise mechanisms by which adherent PMNs may impair endothelial barrier capacity remain unclear. Here, the role of transmembrane signaling by beta(2) integrins in PMN-induced alterations in tight junctional permeability of cultured endothelial cell (EC) monolayers was investigated. PMN activation, in the absence of proinflammatory stimuli, was accomplished through antibody cross-linking of CD11b/CD18, mimicking adhesion dependent receptor engagement. CD18 cross-linking in PMNs added to the EC monolayer provoked a prompt increase in EC permeability that coincided with a rise in EC cytosolic free Ca(2+) and rearrangement of actin filaments, events similar to those evoked by chemoattractant PMN activation. Cell-free supernatant obtained after CD18 cross-linking in suspended PMNs triggered an EC response indistinguishable from that induced by direct PMN activation, and caused clear cut venular plasma leakage when added to the hamster cheek pouch in vivo preparation. The PMN-evoked EC response was specific to beta(2) integrin engagement inasmuch as antibody cross-linking of l-selectin or CD44 was without effect on EC function. Our data demonstrate a causal link between outside-in signaling by beta(2) integrins and the capacity of PMNs to induce alterations in vascular permeability, and suggest a paracrine mechanism that involves PMN derived cationic protein(s) in the cellular crosstalk between PMNs and ECs. PMID- 10839801 TI - Protective immunity to Bordetella pertussis requires both B cells and CD4(+) T cells for key functions other than specific antibody production. AB - To investigate the fundamental nature of protective immunity to Bordetella pertussis, we studied intranasal immunization of adult mice with formalin-fixed B. pertussis (FFBP), followed by aerosol B. pertussis challenge. Mice given two doses of FFBP intranasally completely cleared a subsequent pertussis aerosol challenge from tracheae and lungs (defined as protection), but there was no correlation between levels of specific antibody and clearance of bacteria. Further, transfer of immune serum before aerosol challenge had minimal effects on bacterial burdens. However, pertussis-specific T cells producing interferon gamma but not interleukin 4 or interleukin 10 were detected in draining lymph nodes of FFBP-immunized mice. Significantly, repeated immunization of B cell knockout (BKO) mice resulted in partial protection, and complete protection was reconstituted by transfer of pertussis-immune B cells; reconstituted BKO mice had little if any detectable antipertussis antibodies. Immunization of mice lacking all T cells or lacking CD4(+) T cells did not lead to protection; in contrast, CD8(-) mice were protected. Mice depleted of CD4(+) T cells after immunization but before aerosol challenge, which thus had normal amounts of specific antibodies, were not optimally protected. Taken together, these data indicate that protective immunity to pertussis is dependent on both CD4(+) T cells and B cells, and both cell types provide significant functions other than specific antibody production. PMID- 10839802 TI - Viral escape by selection of cytotoxic T cell-resistant variants in influenza A virus pneumonia. AB - Antigenic variation is a strategy exploited by influenza viruses to promote survival in the face of the host adaptive immune response and constitutes a major obstacle to efficient vaccine development. Thus, variation in the surface glycoproteins hemagglutinin and neuraminidase is reflected by changes in susceptibility to antibody neutralization. This has led to the current view that antibody-mediated selection of influenza A viruses constitutes the basis for annual influenza epidemics and periodic pandemics. However, infection with this virus elicits a vigorous protective CD8(+) cytotoxic T lymphocyte (CTL) response, suggesting that CD8(+) CTLs might exert selection pressure on the virus. Studies with influenza A virus-infected transgenic mice bearing a T cell receptor (TCR) specific for viral nucleoprotein reveal that virus reemergence and persistence occurs weeks after the acute infection has apparently been controlled. The persisting virus is no longer recognized by CTLs, indicating that amino acid changes in the major viral nucleoprotein CTL epitope can be rapidly accumulated in vivo. These mutations lead to a total or partial loss of recognition by polyclonal CTLs by affecting presentation of viral peptide by class I major histocompatibility complex (MHC) molecules, or by interfering with TCR recognition of the mutant peptide-MHC complex. These data illustrate the distinct features of pulmonary immunity in selection of CTL escape variants. The likelihood of emergence and the biological impact of CTL escape variants on the clinical outcome of influenza pneumonia in an immunocompetent host, which is relevant for the design of preventive vaccines against this and other respiratory viral infections, are discussed. PMID- 10839803 TI - T cell receptor-induced calcineurin activation regulates T helper type 2 cell development by modifying the interleukin 4 receptor signaling complex. AB - The activation of downstream signaling pathways of both T cell receptor (TCR) and interleukin 4 receptor (IL-4R) is essential for T helper type 2 (Th2) cell development, which is central to understanding immune responses against helminthic parasites and in allergic and autoimmune diseases. However, little is known about how these two distinct signaling pathways cooperate with each other to induce Th2 cells. Here, we show that successful Th2 cell development depends on the effectiveness of TCR-induced activation of calcineurin. An inhibitor of calcineurin activation, FK506, inhibited the in vitro anti-TCR-induced Th2 cell generation in a dose-dependent manner. Furthermore, the development of Th2 cells was significantly impaired in naive T cells from dominant-negative calcineurin Aalpha transgenic mice, whereas that of Th1 cells was less affected. Efficient calcineurin activation in naive T cells upregulated Janus kinase (Jak)3 transcription and the amount of protein. The generation of Th2 cells induced in vitro by anti-TCR stimulation was inhibited significantly by the presence of Jak3 antisense oligonucleotides, suggesting that the Jak3 upregulation is an important event for the Th2 cell development. Interestingly, signal transducer and activator of transcription (STAT)5 became physically and functionally associated with the IL-4R in the anti-TCR-activated developing Th2 cells that received efficient calcineurin activation, and also in established cloned Th2 cells. In either cell population, the inhibition of STAT5 activation resulted in a diminished IL-4-induced proliferation. Moreover, our results suggest that IL-4 induced STAT5 activation is required for the expansion process of developing Th2 cells. Thus, Th2 cell development is controlled by TCR-mediated activation of the Ca(2+)/calcineurin pathway, at least in part, by modifying the functional structure of the IL-4R signaling complex. PMID- 10839804 TI - Receptor revision of immunoglobulin heavy chain variable region genes in normal human B lymphocytes. AB - Contrary to the general precepts of the clonal selection theory, several recent studies have provided evidence for the secondary rearrangement of immunoglobulin (Ig) genes in peripheral lymphoid tissues. These analyses typically used transgenic mouse models and have only detected secondary recombination of Ig light chain genes. Although Ig heavy chain variable region (V(H)) genes encode a substantial element of antibody combining site specificity, there is scant evidence for V(H) gene rearrangement in the periphery, leaving the physiological importance of peripheral recombination questionable. The extensive somatic mutations and clonality of the IgD(+)Strictly-IgM(-)CD38(+) human tonsillar B cell subpopulation have now allowed detection of the first clear examples of receptor revision of human V(H) genes. The revised VDJ genes contain "hybrid" V(H) gene segments consisting of portions from two separate germline V(H) genes, a phenomenon previously only detected due to the pressures of a transgenic system. PMID- 10839805 TI - In vivo identification of glycolipid antigen-specific T cells using fluorescent CD1d tetramers. AB - The CD1 family of major histocompatibility complex (MHC)-like molecules specializes in presenting lipid and glycolipid antigens to alpha/beta T lymphocytes, but little is known about the size of the CD1-restricted T cell population or the frequency of T lymphocytes specific for a given glycolipid antigen. Here, we report the generation and use of mouse CD1d1-glycolipid tetramers to visualize CD1d-restricted T cells. In contrast with previous BIAcore based estimates of very short half-lives for CD1d-glycolipid complexes, we found that the dissociation rate of several different CD1d-glycolipid complexes was very slow. Fluorescent tetramers of mouse CD1d1 complexed with alpha galactosylceramide (alphaGalCer), the antigen recognized by mouse Valpha14 Jalpha281/Vbeta8 and human Valpha24-JalphaQ/Vbeta11 natural killer T (NKT) cell T cell receptors (TCRs), allowed us for the first time to accurately describe, based on TCR specificity, the entire population of NKT cells in vivo and to identify a previously unrecognized population of NK1.1-negative "NKT" cells, which expressed a different pattern of integrins. In contrast, natural killer (NK) cells failed to bind the tetramers either empty or loaded with alphaGalCer, suggesting the absence of a CD1d-specific, antigen-nonspecific NK receptor. Mouse CD1d1-alphaGalCer tetramers also stained human NKT cells, indicating that they will be useful for probing a range of mouse and human conditions such as insulin dependent diabetes mellitus, tumor rejection, and infectious diseases where NKT cells play an important role. PMID- 10839806 TI - Rho family GTPase Cdc42 is essential for the actin-based motility of Shigella in mammalian cells. AB - Shigella, the causative agent of bacillary dysentery, is capable of directing its movement within host cells by exploiting actin dynamics. The VirG protein expressed at one pole of the bacterium can recruit neural Wiskott-Aldrich syndrome protein (N-WASP), a downstream effector of Cdc42. Here, we show that Cdc42 is required for the actin-based motility of Shigella. Microinjection of a dominant active mutant Cdc42, but not Rac1 or RhoA, into Swiss 3T3 cells accelerated Shigella motility. In add-back experiments in Xenopus egg extracts, addition of a guanine nucleotide dissociation inhibitor for the Rho family, RhoGDI, greatly diminished the bacterial motility or actin assembly, which was restored by adding activated Cdc42. In N-WASP-depleted extracts, the bacterial movement almost arrested was restored by adding exogenous N-WASP but not H208D, an N-WASP mutant defective in binding to Cdc42. In pyrene actin assay, Cdc42 enhanced VirG-stimulating actin polymerization by N-WASP-actin-related protein (Arp)2/3 complex. Actually, Cdc42 stimulated actin cloud formation on the surface of bacteria expressing VirG in a solution containing N-WASP, Arp2/3 complex, and G-actin. Immunohistological study of Shigella-infected cells expressing green fluorescent protein-tagged Cdc42 revealed that Cdc42 accumulated by being colocalized with actin cloud at one pole of intracellular bacterium. Furthermore, overexpression of H208D mutant in cells interfered with the actin assembly of infected Shigella and diminished the intra- and intercellular spreading. These results suggest that Cdc42 activity is involved in initiating actin nucleation mediated by VirG-N-WASP-Arp2/3 complex formed on intracellular Shigella. PMID- 10839807 TI - Effects of in vivo CD8(+) T cell depletion on virus replication in rhesus macaques immunized with a live, attenuated simian immunodeficiency virus vaccine. AB - The role of CD8(+) T lymphocytes in controlling replication of live, attenuated simian immunodeficiency virus (SIV) was investigated as part of a vaccine study to examine the correlates of protection in the SIV/rhesus macaque model. Rhesus macaques immunized for >2 yr with nef-deleted SIV (SIVmac239Deltanef) and protected from challenge with pathogenic SIVmac251 were treated with anti-CD8 antibody (OKT8F) to deplete CD8(+) T cells in vivo. The effects of CD8 depletion on viral load were measured using a novel quantitative assay based on real-time polymerase chain reaction using molecular beacons. This assay allows simultaneous detection of both the vaccine strain and the challenge virus in the same sample, enabling direct quantification of changes in each viral population. Our results show that CD8(+) T cells were depleted within 1 h after administration of OKT8F, and were reduced by as much as 99% in the peripheral blood. CD8(+) T cell depletion was associated with a 1-2 log increase in SIVmac239Deltanef plasma viremia. Control of SIVmac239Deltanef replication was temporally associated with the recovery of CD8(+) T cells between days 8 and 10. The challenge virus, SIVmac251, was not detectable in either the plasma or lymph nodes after depletion of CD8(+) T cells. Overall, our results indicate that CD8(+) T cells play an important role in controlling replication of live, attenuated SIV in vivo. PMID- 10839808 TI - Unexpected rearrangement and expression of the immunoglobulin lambda1 locus in scid mice. AB - In severe combined immunodeficient (scid) mice, V(D)J recombination is severely impaired due to a recessive mutation (scid). Thus, we were surprised to find in this study that Vlambda1-Jlambda1 rearrangement is routinely detectable in scid fetal liver, adult bone marrow, and spleen in the apparent absence of completed VH-DJH and Vkappa-Jkappa rearrangements. Particularly surprising, we found the level of Vlambda1-Jlambda1 rearrangement in scid fetal liver to be comparable to that in fetal liver of wild-type mice. The majority of scid Vlambda1-Jlambda1 rearrangements contained abnormal deletions at the VJ junction, consistent with the known effect of scid. However, approximately 15% of Vlambda1-Jlambda1 rearrangements lacked abnormal deletions. Productive lambda1 transcripts resulting from in-frame rearrangements were readily detectable in scid adult bone marrow and spleen, consistent with our ability to detect lambda1-expressing cells by flow cytometry in the spleens of bcl-2-transgenic scid mice. Strikingly, lambda1 transcripts from individual scid mice often showed VJ junctional sequences with the same recurring palindromic (P) additions of three, four, or five nucleotides. To account for these findings, we suggest that (a) nonhomologous end joining of Vlambda1 and Jlambda1 coding ends in fetal B lineage cells may not be (severely) impaired by scid; (b) recurring P additions in scid lambda1 transcripts may reflect certain molecular constraints imposed by scid on the resolution of Vlambda1 and Jlambda1 hairpin coding ends; and (c), scid lymphocytes with productively rearranged Vlambda1 and Jlambda1 elements may differentiate into recombinase-inactive cells and emigrate from bone marrow to spleen. PMID- 10839809 TI - Enhanced growth of primary tumors in cancer-prone mice after immunization against the mutant region of an inherited oncoprotein. AB - One major objective of tumor immunologists is to prevent cancer development in individuals at high risk. (TG.AC x C57BL/6)F1 mice serve as a model for testing the feasibility of this objective. The mice carry in the germline a mutant ras oncogene that has an arginine at codon 12 instead of glycine present in the wild type, and after physical (wounding) or chemical promotion, these mice have a high probability for developing papillomas that progress to cancer. Furthermore, F1 mice immunized with Arg(12) mutant ras peptide in complete Freund's adjuvant (CFA) develop T cells within 10 d that proliferate in vitro on stimulation with the Arg(12) mutant ras peptide. Within 14 d, these mice have delayed-type hypersensitivity to the peptide. Immunization with CFA alone or with a different Arg(12) mutant ras peptide in CFA induced neither response. To determine the effect of immunization on development of tumors, mice immunized 3 wk earlier were painted on the back with phorbol 12-myristate 13-acetate every 3 d for 8 wk. The time of appearance and the number of papillomas were about the same in immunized and control mice, but the tumors grew faster and became much larger in the mice immunized with the Arg(12) mutant ras peptide. Thus, the immunization failed to protect against growth of papillomas. The peptide-induced CD4(+) T cells preferentially recognized the peptide but not the native mutant ras protein. On the other hand, mice immunized with Arg(12) mutant ras peptide and bearing papillomas had serum antibodies that did bind native mutant ras protein. Together, these studies indicate that active immunization of cancer-prone individuals may result in immune responses that fail to eradicate mutant oncogene expressing tumor cells, but rather induce a remarkable enhancement of tumor growth. PMID- 10839810 TI - Receptor-mediated uptake of antigen/heat shock protein complexes results in major histocompatibility complex class I antigen presentation via two distinct processing pathways. AB - Heat shock proteins (HSPs) derived from tumors or virally infected cells can stimulate antigen-specific CD8(+) T cell responses in vitro and in vivo. Although this antigenicity is known to arise from HSP-associated peptides presented to the immune system by major histocompatibility complex (MHC) class I molecules, the cell biology underlying this presentation process remains poorly understood. Here we show that HSP 70 binds to the surface of antigen presenting cells by a mechanism with the characteristics of a saturable receptor system. After this membrane interaction, processing and MHC class I presentation of the HSP associated antigen can occur via either a cytosolic (transporter associated with antigen processing [TAP] and proteasome-dependent) or an endosomal (TAP and proteasome-independent) route, with the preferred pathway determined by the sequence context of the optimal antigenic peptide within the HSP-associated material. These findings not only characterize two highly efficient, specific pathways leading to the conversion of HSP-associated antigens into ligands for CD8(+) T cells, they also imply the existence of a mechanism for receptor facilitated transmembrane transport of HSP or HSP-associated ligands from the plasma membrane or lumen of endosomes into the cytosol. PMID- 10839811 TI - Cross-presentation of glycoprotein 96-associated antigens on major histocompatibility complex class I molecules requires receptor-mediated endocytosis. AB - Heat shock proteins (HSPs) like glycoprotein (gp)96 (glucose-regulated protein 94 [grp94]) are able to induce specific cytotoxic T lymphocyte (CTL) responses against cells from which they originate. Here, we demonstrate that for CTL activation by gp96-chaperoned peptides, specific receptor-mediated uptake of gp96 by antigen-presenting cells (APCs) is required. Moreover, we show that in both humans and mice, only professional APCs like dendritic cells (DCs), macrophages, and B cells, but not T cells, are able to bind gp96. The binding is saturable and can be inhibited using unlabeled gp96 molecules. Receptor binding by APCs leads to a rapid internalization of gp96, which colocalizes with endocytosed major histocompatibility complex (MHC) class I and class II molecules in endosomal compartments. Incubation of gp96 molecules isolated from cells expressing an adenovirus type 5 E1B epitope with the DC line D1 results in the activation of E1B-specific CTLs. This CTL activation can be specifically inhibited by the addition of irrelevant gp96 molecules not associated with E1B peptides. Our results demonstrate that only receptor-mediated endocytosis of gp96 molecules leads to MHC class I-restricted re-presentation of gp96-associated peptides and CTL activation; non-receptor-mediated, nonspecific endocytosis is not able to do so. Thus, we provide evidence on the mechanisms by which gp96 is participating in the cross-presentation of antigens from cellular origin. PMID- 10839812 TI - Metabolic depletion of ATP by fructose inversely controls CD95- and tumor necrosis factor receptor 1-mediated hepatic apoptosis. AB - Hepatocyte apoptosis is crucial in several forms of liver disease. Here, we examined in different models of murine liver injury whether and how metabolically induced alterations of hepatocyte ATP levels control receptor-mediated apoptosis. ATP was depleted either in primary hepatocytes or in vivo by various phosphate trapping carbohydrates such as fructose. After the activation of the tumor necrosis factor (TNF) receptor or CD95, the extent of hepatocyte apoptosis and liver damage was quantified. TNF-induced cell death was completely blocked in ATP depleted hepatocyte cultures, whereas apoptosis mediated by CD95 was enhanced. Similarly, acute TNF-induced liver injury in mice was entirely inhibited by ATP depletion with ketohexoses, whereas CD95-mediated hepatotoxicity was enhanced. ATP depletion prevented mitochondrial cytochrome c release, loss of mitochondrial membrane potential, activation of type II caspases, DNA fragmentation, and cell lysis after exposure to TNF. The extent of apoptosis inhibition correlated with the severity of ATP depletion, and TNF-induced apoptosis was restored when ATP was repleted by increasing the extracellular phosphate concentration. Our study demonstrates that TNF-induced hepatic apoptosis can be selectively and reversibly blocked upstream of mitochondrial dysfunction by ketohexose-mediated ATP depletion. PMID- 10839813 TI - Modulation of susceptibility to HIV-1 infection by the cytotoxic T lymphocyte antigen 4 costimulatory molecule. AB - CD4 T cells activated in vitro by anti-CD3/28-coated beads are resistant to infection by CC chemokine receptor 5 (CCR5)-dependent HIV-1 isolates. In vivo, antigen-presenting cells (APCs) activate CD4 T cells in part by signaling through the T cell receptor and CD28, yet cells stimulated in this manner are susceptible to HIV-1 infection. We show that cytotoxic T lymphocyte antigen 4 (CTLA-4) engagement counteracts the CD28 antiviral effects, and that the ratio of CTLA-4 to CD28 engagement determines the susceptibility of HIV-1 infection. Furthermore, unopposed CTLA-4 signaling provided by CD28 blockade promotes vigorous HIV-1 replication, despite minimal T cell proliferation. Finally, CTLA-4 antibodies decrease the susceptibility of antigen-activated CD4 T cells to HIV, suggesting a potential approach to prevent or limit viral spread in HIV-1-infected individuals. PMID- 10839814 TI - Fcalpha receptor (CD89) mediates the development of immunoglobulin A (IgA) nephropathy (Berger's disease). Evidence for pathogenic soluble receptor-Iga complexes in patients and CD89 transgenic mice. AB - The pathogenesis of immunoglobulin A (IgA) nephropathy (IgAN), the most prevalent form of glomerulonephritis worldwide, involves circulating macromolecular IgA1 complexes. However, the molecular mechanism(s) of the disease remain poorly understood. We report here the presence of circulating soluble FcalphaR (CD89) IgA complexes in patients with IgAN. Soluble CD89 was identified as a glycoprotein with a 24-kD backbone that corresponds to the expected size of CD89 extracellular domains. To demonstrate their pathogenic role, we generated transgenic (Tg) mice expressing human CD89 on macrophage/monocytes, as no CD89 homologue is found in mice. These mice spontaneously developed massive mesangial IgA deposition, glomerular and interstitial macrophage infiltration, mesangial matrix expansion, hematuria, and mild proteinuria. The molecular mechanism was shown to involve soluble CD89 released after interaction with IgA. This release was independent of CD89 association with the FcRgamma chain. The disease was induced in recombination activating gene (RAG)2(-/-) mice by injection of serum from Tg mice, and in severe combined immunodeficiency (SCID)-Tg mice by injection of patients' IgA. Depletion of soluble CD89 from serum abolished this effect. These results reveal the key role of soluble CD89 in the pathogenesis of IgAN and provide an in vivo model that will be useful for developing new treatments. PMID- 10839815 TI - CD40-CD40 ligand interactions in vivo regulate migration of antigen-bearing dendritic cells from the skin to draining lymph nodes. AB - Whereas CD40-CD40 ligand interactions are important for various dendritic cell (DC) functions in vitro, their in vivo relevance is unknown. We analyzed the DC status of CD40 ligand -/- mice using a contact hypersensitivity (CHS) model system that enables multiple functions of DCs to be assessed in vivo. Immunohistochemistry of skin sections revealed no differences in terms of numbers and morphology of dendritic epidermal Langerhans cells (LCs) in unsensitized CD40 ligand -/- mice as compared with wild-type C57BL/6 mice. However, after contact sensitization of CD40 ligand -/- mice, LCs failed to migrate out of the skin and substantially fewer DCs accumulated in draining lymph nodes (DLNs). Furthermore, very few antigen-bearing DCs could be detected in the paracortical region of lymph nodes draining sensitized skin. This defect in DC migration after hapten sensitization was associated with defective CHS responses and decreased cutaneous tumor necrosis factor (TNF)-alpha production and was corrected by injecting recombinant TNF-alpha or an agonistic anti-CD40 monoclonal antibody. Thus, CD40 CD40 ligand interactions in vivo regulate the migration of antigen-bearing DCs from the skin to DLNs via TNF-alpha production and play a vital role in the initiation of acquired T cell-mediated immunity. PMID- 10839816 TI - Role of antigen-presenting cells in mediating tolerance and autoimmunity. AB - The mechanisms that determine whether receptor stimulation leads to lymphocyte tolerance versus activation remain poorly understood. We have used rat insulin promoter (RIP)-gp/P14 double-transgenic mice expressing the lymphocytic choriomeningitis virus (LCMV) glycoprotein (gp) on pancreatic beta-islet cells together with T cells expressing an LCMV-gp-specific T cell receptor to assess the requirements for the induction of autoimmunity. Our studies have shown that administration of the gp peptide gp33 leads to the activation of P14-transgenic T cells, as measured by the upregulation of activation markers and the induction of effector cytotoxic activity. This treatment also leads to expansion and deletion of P14 T cells. Despite the induction of cytotoxic T lymphocyte activity, peptide administration is not sufficient to induce diabetes. However, the administration of gp peptide together with an activating anti-CD40 antibody rapidly induces diabetes. These findings suggest that the induction of tolerance versus autoimmunity is determined by resting versus activated antigen-presenting cells. PMID- 10839818 TI - Maintenance and integrity of the mitochondrial genome: a plethora of nuclear genes in the budding yeast. AB - Instability of the mitochondrial genome (mtDNA) is a general problem from yeasts to humans. However, its genetic control is not well documented except in the yeast Saccharomyces cerevisiae. From the discovery, 50 years ago, of the petite mutants by Ephrussi and his coworkers, it has been shown that more than 100 nuclear genes directly or indirectly influence the fate of the rho(+) mtDNA. It is not surprising that mutations in genes involved in mtDNA metabolism (replication, repair, and recombination) can cause a complete loss of mtDNA (rho(0) petites) and/or lead to truncated forms (rho(-)) of this genome. However, most loss-of-function mutations which increase yeast mtDNA instability act indirectly: they lie in genes controlling functions as diverse as mitochondrial translation, ATP synthase, iron homeostasis, fatty acid metabolism, mitochondrial morphology, and so on. In a few cases it has been shown that gene overexpression increases the levels of petite mutants. Mutations in other genes are lethal in the absence of a functional mtDNA and thus convert this petite-positive yeast into a petite-negative form: petite cells cannot be recovered in these genetic contexts. Most of the data are explained if one assumes that the maintenance of the rho(+) genome depends on a centromere-like structure dispensable for the maintenance of rho(-) mtDNA and/or the function of mitochondrially encoded ATP synthase subunits, especially ATP6. In fact, the real challenge for the next 50 years will be to assemble the pieces of this puzzle by using yeast and to use complementary models, especially in strict aerobes. PMID- 10839819 TI - Life history and developmental processes in the basidiomycete Coprinus cinereus. AB - Coprinus cinereus has two main types of mycelia, the asexual monokaryon and the sexual dikaryon, formed by fusion of compatible monokaryons. Syngamy (plasmogamy) and karyogamy are spatially and temporally separated, which is typical for basidiomycetous fungi. This property of the dikaryon enables an easy exchange of nuclear partners in further dikaryotic-monokaryotic and dikaryotic-dikaryotic mycelial fusions. Fruiting bodies normally develop on the dikaryon, and the cytological process of fruiting-body development has been described in its principles. Within the specialized basidia, present within the gills of the fruiting bodies, karyogamy occurs in a synchronized manner. It is directly followed by meiosis and by the production of the meiotic basidiospores. The synchrony of karyogamy and meiosis has made the fungus a classical object to study meiotic cytology and recombination. Several genes involved in these processes have been identified. Both monokaryons and dikaryons can form multicellular resting bodies (sclerotia) and different types of mitotic spores, the small uninucleate aerial oidia, and, within submerged mycelium, the large thick-walled chlamydospores. The decision about whether a structure will be formed is made on the basis of environmental signals (light, temperature, humidity, and nutrients). Of the intrinsic factors that control development, the products of the two mating type loci are most important. Mutant complementation and PCR approaches identified further genes which possibly link the two mating type pathways with each other and with nutritional regulation, for example with the cAMP signaling pathway. Among genes specifically expressed within the fruiting body are those for two galectins, beta-galactoside binding lectins that probably act in hyphal aggregation. These genes serve as molecular markers to study development in wild-type and mutant strains. The isolation of genes for potential non-DNA methyltransferases, needed for tissue formation within the fruiting body, promises the discovery of new signaling pathways, possibly involving secondary fungal metabolites. PMID- 10839820 TI - A functional-phylogenetic classification system for transmembrane solute transporters. AB - A comprehensive classification system for transmembrane molecular transporters has been developed and recently approved by the transport panel of the nomenclature committee of the International Union of Biochemistry and Molecular Biology. This system is based on (i) transporter class and subclass (mode of transport and energy coupling mechanism), (ii) protein phylogenetic family and subfamily, and (iii) substrate specificity. Almost all of the more than 250 identified families of transporters include members that function exclusively in transport. Channels (115 families), secondary active transporters (uniporters, symporters, and antiporters) (78 families), primary active transporters (23 families), group translocators (6 families), and transport proteins of ill defined function or of unknown mechanism (51 families) constitute distinct categories. Transport mode and energy coupling prove to be relatively immutable characteristics and therefore provide primary bases for classification. Phylogenetic grouping reflects structure, function, mechanism, and often substrate specificity and therefore provides a reliable secondary basis for classification. Substrate specificity and polarity of transport prove to be more readily altered during evolutionary history and therefore provide a tertiary basis for classification. With very few exceptions, a phylogenetic family of transporters includes members that function by a single transport mode and energy coupling mechanism, although a variety of substrates may be transported, sometimes with either inwardly or outwardly directed polarity. In this review, I provide cross-referencing of well-characterized constituent transporters according to (i) transport mode, (ii) energy coupling mechanism, (iii) phylogenetic grouping, and (iv) substrates transported. The structural features and distribution of recognized family members throughout the living world are also evaluated. The tabulations should facilitate familial and functional assignments of newly sequenced transport proteins that will result from future genome sequencing projects. PMID- 10839821 TI - Type I restriction systems: sophisticated molecular machines (a legacy of Bertani and Weigle). AB - Restriction enzymes are well known as reagents widely used by molecular biologists for genetic manipulation and analysis, but these reagents represent only one class (type II) of a wider range of enzymes that recognize specific nucleotide sequences in DNA molecules and detect the provenance of the DNA on the basis of specific modifications to their target sequence. Type I restriction and modification (R-M) systems are complex; a single multifunctional enzyme can respond to the modification state of its target sequence with the alternative activities of modification or restriction. In the absence of DNA modification, a type I R-M enzyme behaves like a molecular motor, translocating vast stretches of DNA towards itself before eventually breaking the DNA molecule. These sophisticated enzymes are the focus of this review, which will emphasize those aspects that give insights into more general problems of molecular and microbial biology. Current molecular experiments explore target recognition, intramolecular communication, and enzyme activities, including DNA translocation. Type I R-M systems are notable for their ability to evolve new specificities, even in laboratory cultures. This observation raises the important question of how bacteria protect their chromosomes from destruction by newly acquired restriction specifities. Recent experiments demonstrate proteolytic mechanisms by which cells avoid DNA breakage by a type I R-M system whenever their chromosomal DNA acquires unmodified target sequences. Finally, the review will reflect the present impact of genomic sequences on a field that has previously derived information almost exclusively from the analysis of bacteria commonly studied in the laboratory. PMID- 10839823 TI - The SWAL-QOL outcomes tool for oropharyngeal dysphagia in adults: I. Conceptual foundation and item development. AB - In the past two decades, noteworthy advances have been made in measuring the physiologic outcomes of dysphagia, including measurement of duration of structure and bolus movements, stasis, and penetration-aspiration. However, there is a paucity of data on health outcomes from the patients' perspective, such as quality of life and patient satisfaction. A patient-based, dysphagia-specific outcomes tool is needed to enhance information on treatment variations and treatment effectiveness. We present the conceptual foundation and item generation process for the SWAL-QOL, a quality of life and quality of care outcomes tool under development for dysphagia researchers and clinicians. PMID- 10839817 TI - Translational control of viral gene expression in eukaryotes. AB - As obligate intracellular parasites, viruses rely exclusively on the translational machinery of the host cell for the synthesis of viral proteins. This relationship has imposed numerous challenges on both the infecting virus and the host cell. Importantly, viruses must compete with the endogenous transcripts of the host cell for the translation of viral mRNA. Eukaryotic viruses have thus evolved diverse mechanisms to ensure translational efficiency of viral mRNA above and beyond that of cellular mRNA. Mechanisms that facilitate the efficient and selective translation of viral mRNA may be inherent in the structure of the viral nucleic acid itself and can involve the recruitment and/or modification of specific host factors. These processes serve to redirect the translation apparatus to favor viral transcripts, and they often come at the expense of the host cell. Accordingly, eukaryotic cells have developed antiviral countermeasures to target the translational machinery and disrupt protein synthesis during the course of virus infection. Not to be outdone, many viruses have answered these countermeasures with their own mechanisms to disrupt cellular antiviral pathways, thereby ensuring the uncompromised translation of virion proteins. Here we review the varied and complex translational programs employed by eukaryotic viruses. We discuss how these translational strategies have been incorporated into the virus life cycle and examine how such programming contributes to the pathogenesis of the host cell. PMID- 10839824 TI - The SWAL-QOL outcomes tool for oropharyngeal dysphagia in adults: II. Item reduction and preliminary scaling. AB - The SWAL-QOL outcomes tool was constructed for use in clinical research for patients with oropharyngeal dysphagia. The SWAL-QOL was constructed a priori to enable preliminary psychometric analyses of items and scales before its final validation. This article describes data analysis from a pretest of the SWAL-QOL. We evaluated the different domains of the SWAL-QOL for respondent burden, data quality, item variability, item convergent validity, internal consistency reliability as measured by Cronbach's alpha, and range and skewness of scale scores upon aggregation and floor and ceiling effects. The item reduction techniques outlined reduced the SWAL-QOL from 185 to 93 items. The pretest of the SWAL-QOL afforded us the opportunity to select items for the ongoing validation study which optimally met our a priori psychometric criteria of high data quality, normal item distributions, and robust evidence of item convergent validity. PMID- 10839825 TI - An important tool for measuring quality of life. PMID- 10839828 TI - Structural displacements in normal swallowing: a videofluoroscopic study. AB - Dynamic videofluoroscopic swallow studies were performed on 60 normal adult volunteers to establish normative data for displacement of upper aerodigestive tract structures during deglutition. Variables evaluated included hyoid bone displacement, larynx-to-hyoid bone approximation, pharyngeal constriction, and the extent of pharyngoesophageal sphincter (PES) opening during liquid swallows of 1, 3, and 20 cc. Results showed direct relationships between bolus size and hyoid displacement, between bolus size and PES opening, and between bolus size and pharyngeal constriction. Only hyoid-to-larynx approximation remained unchanged across bolus sizes. Sex differences were noted for all variables except PES opening. Reliability for most measurement variables was excellent. To our knowledge, normative data for pharyngeal constriction and larynx-to-hyoid approximation have not previously been described. PMID- 10839827 TI - Effects of three techniques on maximum posterior movement of the tongue base. AB - Effects of three techniques designed to improve maximum range of posterior movement of the base of tongue were investigated under videofluoroscopy in 20 subjects. Retraction of the tongue base during 3-ml pudding swallows, tongue pull back, yawn, and gargle tasks was measured in millimeters, with the second cervical vertebra as a reference point and was judged subjectively as well. The gargle task was the most successful in eliciting most tongue base retraction for the group of subjects, although not in every subject. Gargle also resulted in greater tongue base movement than swallow more often than the other two voluntary tasks. Clinicians' subjective judgment of "most" retracted tongue base position was generally reliable in comparison with actual measurements. The number of repeat swallows on each bolus correlated significantly with the approximate percentage of residue in the valleculae as judged clinically. PMID- 10839826 TI - Clinical utility of the modified barium swallow. AB - The purpose of this investigation was to evaluate the immediate and clinically relevant information gained from the modified barium swallow study and to determine the impact of the procedure on patient management. A database containing a nonrandom sample of 608 swallowing studies was reviewed. Results showed that only 10.4% of the studies were classified as normal examinations and aspiration occurred in 32.4%. However, swallowing abnormality without aspiration was recorded in 57.2% of the studies. Five additional outcome variables were assessed: referrals made to other specialties, effectiveness of applied compensatory strategies, treatment recommendations, mode of intake change, and diet grade change. Nearly 83% of the 608 studies showed change in at least one of the variables: needed referral to a specialist was identified on 26.3%; compensatory strategies that improved swallow physiology were identified on 48.4%; swallowing therapy was recommended on 37.2%; changes in mode of intake occurred on 31.4%; and diet texture changes were recommended on 43.8%. The low percentage of normal studies coupled with the high percentage of change in measurable variables indicate high clinical utility for the modified barium swallow study. The misguided tendency to refer to the modified barium study only as a tool for identifying aspiration and the appropriate utilization of the examination for identification of underlying abnormality in swallowing physiology are explained. PMID- 10839829 TI - Deep laryngeal penetration as a predictor of aspiration. AB - Very little has been written about laryngeal penetration as a separate diagnostic event from aspiration. Laryngeal penetration has been described as an infrequent event in adult individuals without swallowing problems. This study describes the incidence of laryngeal penetration in 125 dysphagic children ranging in age from 7 days to 19 years who were seen over a 6-month period at The Children's Hospital in Denver, Colorado. Laryngeal penetration was identified in 60% of the study group, with 31% demonstrating deep laryngeal penetration. Of the children exhibiting deep laryngeal penetration, 85% aspirated, suggesting a strong correlation between these two events. It was noted that children exhibiting deep laryngeal penetration often began to aspirate further into their feedings. Use of extended feedings during videofluoroscopy is discussed as a diagnostic strategy in the presence of deep laryngeal penetration. PMID- 10839830 TI - Swallowing apraxia: a disorder of the Praxis system? AB - The purpose of this review is to evaluate the disorder of swallowing apraxia and determine how it fits into the praxis system. Swallowing apraxia, a proposed disorder of lingual, labial, and mandibular coordination, has been observed before bolus transfer during the oral stage of swallowing. Although frequently discussed anecdotally in dysphagia literature, the possible mechanisms and neural networks of swallowing apraxia have not been elucidated. Similarities and differences of swallowing apraxia with buccofacial, speech, and limb apraxias are evident. Critical review of the literature has identified possible similarities as greater occurrence upon command, transitive nature of the action, and evidence of spatial errors. Conversely, differences such as hemispheric lateralization and multiple gesture assessment may exist between swallowing apraxia and more traditional forms of apraxia. Until discrete error patterns of swallowing apraxia are identified and precisely measured, the nature of this disorder and its relationship with the praxis system will continue to remain elusive. PMID- 10839833 TI - Leadership: A special "dedication" PMID- 10839822 TI - Acetylation of histones and transcription-related factors. AB - The state of chromatin (the packaging of DNA in eukaryotes) has long been recognized to have major effects on levels of gene expression, and numerous chromatin-altering strategies-including ATP-dependent remodeling and histone modification-are employed in the cell to bring about transcriptional regulation. Of these, histone acetylation is one of the best characterized, as recent years have seen the identification and further study of many histone acetyltransferase (HAT) proteins and their associated complexes. Interestingly, most of these proteins were previously shown to have coactivator or other transcription-related functions. Confirmed and putative HAT proteins have been identified from various organisms from yeast to humans, and they include Gcn5-related N-acetyltransferase (GNAT) superfamily members Gcn5, PCAF, Elp3, Hpa2, and Hat1: MYST proteins Sas2, Sas3, Esa1, MOF, Tip60, MOZ, MORF, and HBO1; global coactivators p300 and CREB binding protein; nuclear receptor coactivators SRC-1, ACTR, and TIF2; TATA binding protein-associated factor TAF(II)250 and its homologs; and subunits of RNA polymerase III general factor TFIIIC. The acetylation and transcriptional functions of these HATs and the native complexes containing them (such as yeast SAGA, NuA4, and possibly analogous human complexes) are discussed. In addition, some of these HATs are also known to modify certain nonhistone transcription related proteins, including high-mobility-group chromatin proteins, activators such as p53, coactivators, and general factors. Thus, we also detail these known factor acetyltransferase (FAT) substrates and the demonstrated or potential roles of their acetylation in transcriptional processes. PMID- 10839831 TI - Esophageal biopsy does not predict clinical outcome after percutaneous endoscopic gastrostomy in children. AB - Clinically symptomatic gastroesophageal reflux may occur after percutaneous endoscopic gastrostomy (PEG). Preoperative evaluation for gastroesophageal reflux does not reliably predict those individuals who will develop reflux unresponsive to medical management after PEG. Esophageal histology at the time of PEG might be used to identify patients at risk for developing intractable gastroesophageal reflux. The study aim was to correlate the clinical outcome after PEG with esophageal histology at the time of PEG insertion. A retrospective review of 68 consecutive children who had an esophageal biopsy obtained at the time of PEG insertion was undertaken. Preoperative evaluation, esophageal histology, and clinical outcomes were compared. Preoperative gastroesophageal reflux was present in 23% of upper gastrointestinal series performed, in 10% of pH probe studies, and in 29% of reflux scans. Histology was normal in 57% of esophageal biopsies obtained at the time of PEG insertion. Symptomatic gastroesophageal reflux requiring antireflux surgery or conversion to gastrojejunostomy developed in 10% of patients after PEG placement. Only one of these patients had esophagitis on biopsy. In conclusion, preoperative esophageal histology does not reliably predict the development of symptomatic gastroesophageal reflux after PEG placement. PMID- 10839835 TI - Telephone triage: protocols and clinical judgment are not mutually exclusive PMID- 10839834 TI - On mandatory overtime and wearing blue ribbons. PMID- 10839838 TI - More on successful use of LET for wound repair PMID- 10839837 TI - More on international emergency nursing PMID- 10839839 TI - Pagers successfully reduce ED noise PMID- 10839841 TI - More on ambulatory payment classifications PMID- 10839840 TI - Eliminating triage a very bad idea PMID- 10839843 TI - More on ambulatory payment classifications PMID- 10839845 TI - What happens to "antique" nurses? PMID- 10839846 TI - Fatal aspiration of a cheese cube by a 2-year-old boy. PMID- 10839847 TI - Attitudes of emergency nurses regarding patient assaults. AB - INTRODUCTION: The purpose of this study was to investigate the frequency of patient assaults on emergency nurses and to explore nurses' attitudes about patient assaults. METHOD: A convenience sample of ED nurses (N = 55) from 2 hospitals in the mid-south region of the United States was used. All subjects completed a 31-item questionnaire that assessed frequency of assaults and attitudes about patient assaults. RESULTS: Eighty-two percent of nurses surveyed had been assaulted during their careers. In the preceding year, 56% of nurses had been assaulted; 29% of these assaults were unreported. The majority of nurses (73%) believed that being assaulted "goes with the job." Only 2 nurses (3.6%) felt safe from the possibility of patient assault at work "all of the time." DISCUSSION: The high rate of patient assault and the subsequent underreporting of patient assault identified in our study are consistent with rates reported by other researchers. It is alarming that, although many of these other studies were conducted 10 or more years ago, the assault rates are nearly the same. Of the 45 nurses in this study who had been the victim of patient assaults, only 9 believed that reporting their assaults had been beneficial. Lack of support for nursing, whether it be from other nurses, management, the institution, or society, appears to be responsible for the vast underreporting of patient assaults. Nurses themselves are also unsure of how to proceed. Whereas 91% of the surveyed nurses stated that they believed they had a right to take legal action against an assaultive patient, only 65% stated that it was ethically appropriate and even less (33%) stated that they would actually press charges against an assaultive patient. PMID- 10839848 TI - Usefulness of the modified 0-10 Borg scale in assessing the degree of dyspnea in patients with COPD and asthma. AB - INTRODUCTION: Rapid assessment and monitoring is essential for patients with acute bronchospasm. However, tools for measuring dyspnea or the state of being short of breath are often limited to peak flow, blood gas analysis, and asking patients multiple questions about their breathing at a time when they find speaking difficult. We thus decided to examine a tool called the modified Borg scale (MBS) that had the potential to provide quick, easy, and rapid information about a patient's subjective state of dyspnea. This 0 to 10 rated scale gave our ED patients a device they could use to measure and evaluate their dyspnea. For this reason, we added it to the triage assessment practice and included it in all posttreatment assessment notes on patients with exacerbations of asthma or chronic obstructive pulmonary disease (COPD) who were seen in the emergency department and urgent care clinic. STUDY QUESTIONS: (1) Can patients with acute bronchospastic asthma or COPD adequately communicate their level of dyspnea using the MBS? (2) Does subjective improvement in the patient's dyspnea using the MBS correlate with improvements in pulmonary functions as measured by the peak flow meter and cutaneous oxygen saturation (Sao(2))? METHODS: Routine and triage assessment of subjective dyspnea using the MBS was instituted at a hospital emergency department serving adult veterans. Concurrently, the MBS was added to our standardized treatment protocol for management of patients with bronchospasm. ED and urgent care records were reviewed to collect baseline and postrespiratory treatment data on peak expiratory flow rates (PEFR), MBS scores, and Sao(2) percentages. RESULTS: Four hundred male veterans aged 24 to 87 years presented with a chief complaint of dyspnea. The assessing physician identified 102 of these patients as having acute bronchospasm; 42 were diagnosed with asthma, and 60 were diagnosed with COPD. All study patients with acute bronchospasm were able to use the MBS to rate their perception of severity of dyspnea. As the peak flow measurements increased, the MBS scores of difficulty breathing decreased. For the asthma groups, the mean MBS score decreased from 5.1 at triage baseline to 2.4 after treatment. This finding indicated that a significant correlation existed between the change in MBS scores and the change in PEFR from pretreatment to posttreatment scores (r = -.31, P <.05). As the peak flow increased, the MBS scores decreased. Sao(2) only slightly improved in the asthma group compared with the COPD group. For patients with COPD, the mean MBS score decreased from 6.0 at triage baseline to 3.0 after treatment. This finding indicated that a significant correlation also existed between the change in MBS scores and the change in PEFR from pretreatment to posttreatment scores (r = -.42, P <.001). Cutaneous oxygen saturation also improved in the COPD group after treatment. The modality of treatment ordered by the physician was metered dose inhaler or nebulizer. These treatment modalities had no effect on the aforementioned results in the asthma or COPD group. CONCLUSIONS: The MBS is a valid and reliable assessment tool for dyspnea. This study demonstrated that it correlated well with other clinical parameters and could be useful when assessing and monitoring outcomes in patients with acute bronchospasm. Patients who used the MBS rated it with a high degree of satisfaction on ease of use and found that the language in this scale adequately expressed their dyspnea. The ED triage and primary care nursing staff rated the MBS as highly satisfactory, stating that it was quick and easy to use. Respiratory assessment in the triage notes and nursing notes were streamlined to consistently include 3 respiratory measures: PEFR, MBS, and Sao(2). Long respiratory narratives were found to be unnecessary in many cases. In addition, the MBS helped to include an important element of subjective assessment when evaluating the severity of dyspnea. PMID- 10839849 TI - Reducing risk with telephone follow-up of patients who leave against medical advice or fail to complete an ED visit. PMID- 10839850 TI - Can a self-rating 0-10 scale for dyspnea yield a common language that is understood by ED nurses, patients, and their families? PMID- 10839851 TI - Every now and then...a pat on the back: one emergency department's attempt to improve morale and patient satisfaction. PMID- 10839852 TI - Clinical nurses forum PMID- 10839853 TI - Outcomes of prehospital care: do we really make a difference? PMID- 10839854 TI - Using or misusing the Internet at work. PMID- 10839856 TI - Managers forum PMID- 10839855 TI - Just "sleeping it off". PMID- 10839857 TI - Fine dining or fast food? Setting the table with an effective speaker introduction. PMID- 10839858 TI - Pediatric immunizations: are you up-to-date? PMID- 10839859 TI - Research with elderly ED patients: do we have any answers? PMID- 10839860 TI - A 16-year-old with abdominal pain. PMID- 10839861 TI - Seven strategies to decrease the risk of missed injuries. PMID- 10839862 TI - A 64-year-old woman with a swollen and painful leg. PMID- 10839863 TI - Paydirt. PMID- 10839864 TI - The chronic disease of childhood obesity: the sleeping giant has awakened PMID- 10839865 TI - Helicobacter pylori infection in children: to test or not to test...what is the evidence? PMID- 10839866 TI - Clinical approach to inhaled nitric oxide therapy in the newborn with hypoxemia. PMID- 10839867 TI - Prevalence of abnormal serum aminotransferase values in overweight and obese adolescents. AB - OBJECTIVES: (1) To determine the prevalence of abnormal liver enzymes in overweight and obese adolescents and (2) to determine the relationship of alcohol ingestion and serum antioxidants to the presence of abnormal liver enzymes in overweight and obese adolescents. METHODS: Serum alanine aminotransferase (ALT) and gamma-glutamyl transpeptidase levels were measured in 2450 children between the ages of 12 and 18 years, enrolled in the National Health and Examination Survey, cycle III (NHANES III). Obesity was defined as a body mass index >95th percentile for age and sex. Overweight was defined as a body mass index >85th percentile for age and sex. Nutritional intake was assessed by 24-hour dietary recall and food frequency questionnaires. Serum antioxidants were measured by high-pressure liquid chromatography. RESULTS: Sixty percent of adolescents with elevated ALT levels were either overweight or obese. Overall, 6% of overweight adolescents had elevated ALT levels (odds ratio: 3.4 [95% CI: 3.5-12.8]). Ten percent of obese adolescents had elevated ALT levels (odds ratio: 6.7 [95% CI: 3.5-12.8]). In addition, approximately 1% of obese adolescents demonstrated ALT levels over twice normal. Approximately 50% of of obsese adolescents who reported modest alcohol ingestion (4 times per month or more) had elevated ALT levels (odds ratio: 10.8, 95% CI: 1.5-77). Other factors associated with elevated ALT levels in overweight and obese adolescents include increased age, elevated glycolated hemoglobin, elevated triglycerides, and decreased levels of serum antioxidants-vitamin E, beta-carotene, and vitamin C. CONCLUSION: Overweight and obesity are the most common findings in adolescents with elevated ALT levels. Even modest alcohol consumption may significantly increase the likelihood of obese adolescents developing obesity-related liver disease. PMID- 10839868 TI - Vitamin E treatment of nonalcoholic steatohepatitis in children: a pilot study. AB - AIM: To determine whether supplemental oral vitamin E is effective in lowering serum aminotransferase and alkaline phosphatase levels in children with nonalcoholic steatohepatitis (NASH) associated with obesity. STUDY DESIGN: Open label pilot study enrolling all children <16 years old with chronically elevated serum aminotransferase (alanine aminotransferase and aspartate aminotransferase) levels for greater than 3 months, who demonstrated a diffusely echogenic liver on ultrasonography, had no demonstrable reason for abnormal serum chemistry values other than obesity, and therefore were diagnosed to have NASH. Patients were prescribed oral vitamin E between 400 and 1200 IU per day. Serum chemistry values were monitored monthly during treatment. RESULTS: Eleven subjects with a mean age of 12.4 years were enrolled; treated patients were followed up for 4 to 10 months. The body mass index did not change significantly before and after treatment (32.8 +/- 3.8 kg/m(2) vs 32.5 +/- 4.4 kg/m(2), respectively). Serum alanine aminotransferase decreased from 175 +/- 106 IU/L to 40 +/- 26 IU/L (P <.001, paired Student t test), serum aspartate aminotransferase decreased from 104 +/- 61 IU/L to 33 +/- 11 IU/L (P <.002), and alkaline phosphatase decreased from 279 +/- 42 IU/L to 202 +/- 66 IU/L (P <.003) during treatment. Serum aminotransferase levels remained normal during treatment but returned to abnormal in those electing to stop treatment. Serum alpha-tocopherol levels were within the normal range before the commencement of therapy and increased significantly with supplementation. The liver remained diffusely echogenic during therapy, at the time serum aminotransferase levels were reduced. CONCLUSIONS: Daily oral vitamin E administration normalized serum aminotransferase and alkaline phosphatase levels in children with NASH. Obese children with NASH should be encouraged to lose weight as part of a comprehensive weight reduction program and to consider taking supplemental alpha-tocopherol. PMID- 10839869 TI - Lack of efficacy of ursodeoxycholic acid for the treatment of liver abnormalities in obese children. AB - OBJECTIVE: To determine whether ursodeoxycholic acid (UDCA) is effective for treatment of obesity-related liver abnormalities in children. STUDY DESIGN: Thirty-one children (21 bboys; mean age, 8.7 years) had obesity-related persistent elevation of aminotransferase levels, which was associated with ultrasonographic images of bright liver in 27. A preliminary interview determined which patients were (n = 18) or were not (n = 13) likely to comply with a balanced low-calorie diet. Four subgroups emerged: patients who followed the diet (n = 11), patients treated with UDCA (10 mg/kg/d) given alone (n = 7) or added to the diet (n = 7), and untreated control patients (n = 6). RESULTS: Diet alone determined weight loss and resolved biochemical liver abnormalities in all patients. Addition of UDCA to the diet was no more efficacious than weight loss alone. UDCA alone was ineffective for the treatment of liver abnormalities in all cases, and results did not differ from those observed in the untreated control group. Improvement of ultrasonographic abnormalities was observed in patients who lost weight, irrespective of UDCA administration. CONCLUSIONS: UDCA is not effective for the treatment of obesity-related liver abnormalities in children. PMID- 10839870 TI - Acquisition of Helicobacter pylori infection in a high-risk population occurs within the first 2 years of life. AB - OBJECTIVE: To elucidate age at acquisition of Helicobacter pylori infection, we conducted a study in a population of children known to be at high risk for H pylori infection. METHOD: Children with no symptoms who were of Turkish nationality and living in the city of Ulm, Germany, and nearby communities and on whom routine health screening examinations were performed at age 1 year (56 children), 2 years (55 children), or 4 years (69 children) were included in this cross-sectional study. Stool samples were collected by the parents and sent by mail to the University of Ulm. An H pylori antigen enzyme immunoassay for the detection of H pylori in stool was used to define current infection status. Sensitivity was 84.6% (95% CI 63. 1% to 94.7%) and specificity 97.7% (95% CI 86.2% to 99.9%) in the 4-year-old children in whom the stool test was compared with the (13)C-urea breath test. RESULTS: The prevalence of infection was 8. 9% (95% CI 3.0% to 19.6%) among the 1-year-old children, 36.4% (95% CI 23.8% to 50.4%) among the 2-year-old children, and 31.9% (95% CI 21.2% to 44.2%) among the 4-year-old children. CONCLUSION: In this high-risk group of Turkish children living in Germany, H pylori acquisition seems to occur mainly between the first and the second years of life. Therefore preventive measures such as vaccination necessitate application early in infancy. Further studies are now required to ascertain the mechanisms for transmission in this age group. PMID- 10839871 TI - Prevalence and clinical correlates of glomerulopathy in children with sickle cell disease. AB - OBJECTIVES: Glomerular disease and renal failure cause substantial morbidity for patients with sickle cell disease (SCD). Proteinuria is an early manifestation of sickle nephropathy, but the prevalence of proteinuria and its clinical correlations in children with SCD are unknown. STUDY DESIGN: Data were collected prospectively on children with SCD for 10 years including physical measurements, laboratory test results, and clinical complications. Persistent proteinuria was defined as > or =1+ protein on urinalysis for at least 6 months. The glomerular filtration rate was estimated with serum creatinine concentration and height. Proteinuria was correlated with other variables by chi(2) analysis. RESULTS: Proteinuria occurred in 20 of 442 pediatric patients including 15 (6.2%) with sickle cell anemia. Proteinuria increased with age, affecting 12% of older teenagers with sickle cell anemia. Proteinuria was significantly associated with lower hemoglobin concentration, higher mean corpuscular volume, and higher leukocyte count. For children of some ages, proteinuria was associated with complications including stroke, acute chest syndrome, cholelithiasis, and hospitalizations. Glomerular filtration rate hyperfiltration occurred early in life, followed by normalization. CONCLUSIONS: Sickle nephropathy, manifested as persistent proteinuria, begins early in life, occurs in all forms of SCD, and is associated with severity of disease. Early detection of proteinuria may allow therapy to prevent progressive renal insufficiency. PMID- 10839872 TI - Urodynamic dysfunction in infants with vesicoureteral reflux. AB - PATIENTS AND METHODS: Vesicoureteral reflux (VUR) may result from inadequate length of the intravesical ureteric tunnel and from urodynamic dysfunction. VUR and symptomatic urinary tract infection (UTI) are both more common in male infants and may be related to urodynamic dysfunction and functional intravesical obstruction. We therefore conducted urodynamic studies in 75 infants with primary VUR and retrospectively evaluated their voiding cystourethrogram (VCUG) for evidence of inadequate sphincteric relaxation. RESULTS: High voiding detrusor pressure (>70 cm H(2)O), increased postvoid residual (>2 mL/kg body weight), or detrusor-sphincter dyssynergy were observed in 79% of infants with primary VUR without significant differences between male and female infants or in infants with or without UTI. Infants with bilateral grade IV-V VUR showed a higher prevalence of detrusor hyper-reflexia and higher voiding detrusor pressure than infants with lower grades of VUR or unilateral grade IV-V VUR. On VCUG, inadequate external urethral sphincter relaxation during voiding was noted in 10 of 47 male infants. CONCLUSIONS: The findings suggest that high intravesical pressures contribute to the development and severity of VUR, and that high voiding detrusor pressure in some infants with primary VUR may be related to inadequate relaxation of the external urethral sphincter. PMID- 10839873 TI - Vascular retinal abnormalities in neonates of mothers who smoked during pregnancy. AB - OBJECTIVE: To investigate whether maternal smoking during pregnancy causes retinal abnormalities in the newborn. STUDY DESIGN: One hundred sixty-two neonates of smoking mothers and 162 matched neonates of nonsmoking mothers (112 appropriate for gestational age [AGA], 30 small for gestational age [SGA], 20 large for gestational age [LGA] in each group) were studied. RESULTS: Retinal arterial narrowing and straightening (RANS) was observed in 52 and 10 eyes of the newborns of smoking and nonsmoking mothers, respectively (P <. 000001) in association with elevated blood pressure in the neonates. The frequency of RANS was more than 3-fold greater in the SGA neonates than in the AGA and LGA neonates of the smoking mothers. Retinal venous dilatation and tortuosity (RVDT) was found in 100 and 36 eyes of neonates of smoking and nonsmoking mothers, respectively (P <.000001). The frequency of RVDT in the SGA neonates of the smoking mothers was 2.5-fold and 4.2-fold greater than in the AGA infants and the LGA infants, respectively. Also, intraretinal hemorrhages were found in 61 and 31 eyes of neonates of smoking and nonsmoking mothers, respectively (P =.0007) in association with elevated hematocrit and RVDT, whereas no intraretinal hemorrhages were found when RANS was present. All retinal abnormalities resolved by 6 months in infants of smoking mothers and by 2 months in infants of nonsmoking mothers. CONCLUSIONS: Maternal smoking during pregnancy causes increased frequency of RANS, RVDT, and intraretinal hemorrhages; but these retinal abnormalities resolve by 6 months of age. PMID- 10839874 TI - Gestational age and birth weight effects on plasma clearance of fentanyl in newborn infants. AB - OBJECTIVE: To provide a rational basis for the dosage of fentanyl in newborn infants by determining clearance in the first days of life. STUDY DESIGN: A continuous infusion of fentanyl for 2 to 3 days (10. 5 microg/kg over a 1-hour period followed by 1.5 microg/kg/h) was administered to 38 newborn infants who had undergone ventilation (gestational ages 26 to 42 weeks and birth weights 835 to 3550 g). Fentanyl concentrations were measured in arterial blood samples collected at 2, 12, 24, 48, and 60 hours after the start of fentanyl infusion. Fentanyl levels were correlated with a pain score. RESULTS: The mean (+/-SD) steady-state fentanyl concentration of 2.5 (+/-1) ng/mL achieved between 24 and 48 hours of infusion correlated significantly with the concomitant pain score (r = -0.57, P <.01). The clearance, 11.5 (+/-4.0) mL/min/kg, correlated significantly with the gestational age (r = 0.46, P <.01) and birth weight (r = 0. 48, P <.01). CONCLUSIONS: Because plasma fentanyl clearance increases with maturity, gestational age should be taken into account when fentanyl is administered to newborn infants. PMID- 10839876 TI - The influence of a pacifier on infants' arousals from sleep. AB - OBJECTIVE: The risk of sudden infant death during sleep was postulated to decrease with the use of a pacifier and by conditions facilitating arousals from sleep. We evaluated the influence of a pacifier on arousal from sleep in healthy infants. STUDY DESIGN: Fifty-six healthy infants were studied with polygraphy during 1 night; 36 infants used a pacifier regularly during sleep, and 20 never used a pacifier. Thumb users or occasional pacifier users were not included in the study. The infants were recorded at a median age of 10 weeks (range 6 to 19 weeks). To evaluate auditory arousal thresholds, the infants were exposed to white noise of increasing intensity during rapid eye movement sleep. RESULTS: Polygraphic arousals occurred at significantly lower auditory stimuli in pacifier users than in nonusers (P =.010). Compared with nonusers, pacifier users were more frequently bottle-fed than breast-fed (P =. 036). Among infants sleeping without a pacifier, breast-fed infants had lower auditory thresholds than bottle fed infants (P =.049). CONCLUSIONS: Infants using pacifiers during sleep had lower auditory arousal thresholds than those who did not use a pacifier during sleep. Breast-feeding could be a further factor contributing to lower arousal thresholds. These findings could be relevant to the occurrence of sudden infant deaths during sleep. PMID- 10839875 TI - A new blue light-emitting phototherapy device: a prospective randomized controlled study. AB - OBJECTIVE: To evaluate the efficacy of a new phototherapy light source with a narrow luminous blue spectrum. The device, made with high-intensity gallium nitride light-emitting diodes (LEDs), was compared with conventional phototherapy at similar light intensities. SETTING: Two university-affiliated community hospitals in Jerusalem. DESIGN: Prospective open randomized study. PARTICIPANTS: Sixty-nine jaundiced, but otherwise healthy, term infants who met the entry criteria for phototherapy set by the American Academy of Pediatrics' Practice Parameter. MAIN OUTCOME MEASURES: The duration of phototherapy and the rate of decrease in total serum bilirubin (TSB) concentration. RESULTS: The mean TSB concentrations at initiation and termination of treatment did not differ between newborns receiving LED and those receiving conventional phototherapy. The duration of phototherapy and the rate of decrease in TSB concentration were not statistically different in the 2 groups. The average rate of decrease in TSB after adjustment by a linear regression analysis for confounding factors was 3.16 micromol/L/h (95% confidence limits -4.81, -1.51) in newborns receiving LED phototherapy compared with -2.19 micromol/L/h (-3.99, -0.40) in those treated with conventional phototherapy (P <.14). No side effects were noted in any of the newborns. CONCLUSIONS: The blue gallium nitride LED device is as effective as conventional phototherapy and is readily accepted by nursing staff. Future LED phototherapy devices can provide much higher irradiance, and thus greater efficacy, and offer a new highly versatile approach to the treatment of jaundice. PMID- 10839877 TI - Clinical and virologic response to combination treatment with indinavir, zidovudine, and lamivudine in children with human immunodeficiency virus-1 infection: a multicenter study in the Netherlands. On behalf of the Dutch Study Group for Children with HIV-1 infections. AB - OBJECTIVE: To evaluate the clinical, immunologic, and virologic response to indinavir, zidovudine, and lamivudine in children with human immunodeficiency virus-1 (HIV-1) infection. STUDY DESIGN: Twenty-eight HIV-1-infected children (3 months to 16 years of age) with or without prior treatment with reverse transcriptase inhibitors and a HIV-1 RNA >5000 copies/mL and/or a CD4 cell count less than the lower limit of the age-specific reference value were treated with indinavir, zidovudine, and lamivudine. Pharmacokinetics of indinavir were determined in each child. RESULTS: The combination treatment was well tolerated in the majority of patients. Clinical improvement was seen in all patients. After 6 months of therapy, 70% of the patients had an HIV-1 RNA load below 500 copies/mL, whereas 48% of the children had a viral load below 40 copies/mL. Relative CD4 cell counts in relation to the lower limit of the age-specific reference value increased significantly from a median value of 79% at baseline to 106% after 6 months of therapy. The doses of indinavir necessary to achieve area under the curve values comparable to adult values varied from 1250 mg/m(2)/d to 2450 mg/m(2)/d. CONCLUSIONS: Highly active antiretroviral therapy consisting of indinavir, zidovudine, and lamivudine in children reduced HIV-1 RNA to less than 500 copies/mL in 70% of the children within 6 months. Improved CD4 cell counts were observed in most patients, as was a better clinical condition (no invasive or opportunistic infections, increased weight gain). Side effects of the triple therapy were mild. Highly active antiretroviral therapy can be used as successfully in children as in adults. PMID- 10839879 TI - Liver transplantation and chemotherapy for hepatoblastoma and hepatocellular cancer in childhood and adolescence. AB - OBJECTIVE: To describe our experience with total hepatectomy and liver transplantation as treatment for primary hepatoblastoma (HBL) and hepatocellular carcinoma (HCC) in children. STUDY DESIGN: A retrospective analysis of the perioperative course of 31 children with unresectable primary HBL (n = 12) and HCC (n = 19) who underwent transplantation between May 1989 and December 1998. Systemic (n = 18) and intraarterial (n = 7) neoadjuvant chemotherapy were administered; follow-up ranged from 1 to 185 months. RESULTS: For HBL, 1-year, 3 year, and 5-year posttransplantation survival rates were 92%, 92%, and 83%, respectively. Intravenous invasion, positive hilar lymph nodes, and contiguous spread did not have a significant adverse effect on outcome; distant metastasis was responsible for 2 deaths. Intraarterial chemotherapy was effective in all patients treated. For HCC, the overall 1-year, 3-year, and 5-year disease-free survival rates were 79%, 68%, and 63%, respectively. Vascular invasion, distant metastases, lymph node involvement, tumor size, and gender were significant risk factors for recurrence. Intraarterial chemotherapy was effective in 1 of 3 patients. Six patients died of recurrent HCC, and 3 deaths were unrelated to recurrent tumor. CONCLUSION: Liver transplantation for unresectable HBL and HCC can be curative. Risk factors for recurrence were significant only for HCC, with more advanced stages amenable to cure in the HBL group. PMID- 10839880 TI - Risk of gastrointestinal bleeding during adolescence and early adulthood in children with portal vein obstruction. AB - OBJECTIVE: To estimate the risk of bleeding during adolescence and early adulthood in a group of children with portal vein obstruction who had not undergone an effective treatment beforehand. STUDY DESIGN: Children (n = 44) were followed up from age 12 years to a mean age of 20 years (range, 15-34 years). Actuarial risk of bleeding, related to previous occurrence of gastrointestinal bleeding and to pattern of varices at age 12, was calculated yearly. RESULTS: Twenty-four children presented with gastrointestinal bleeding after age 12, and 20 did not bleed. The overall actuarial probability of bleeding was 49% at age 16 and 76% at age 24. Probability of bleeding at age 23 was higher in children who had bled before age 12 than in children who had not bled (93% vs 56%; P =.007). Probabilities of bleeding at age 18 and at age 23 were 60% and 85%, respectively, in patients who had grade II or III esophageal varices at age 12. The 9 children without varices or with grade I varices only on endoscopy did not bleed between the ages of 12 and 20 years. CONCLUSIONS: Children with portal vein obstruction have a >50% risk of bleeding during adolescence; the pattern of varices on endoscopy at age 12 may have a prognostic value. PMID- 10839878 TI - Lidocaine-prilocaine patch decreases the pain associated with the subcutaneous administration of measles-mumps-rubella vaccine but does not adversely affect the antibody response. AB - BACKGROUND: Topical lidocaine 2.5% and prilocaine 2.5% (EMLA) is effective in decreasing the pain associated with minor procedures including immunization, although the effect on the antibody response to vaccine constituents has not been assessed. OBJECTIVE: To measure the antibody response to measles-mumps-rubella (MMR) vaccine, as well as pain reduction associated with the use of the EMLA patch. PARTICIPANTS AND SETTING: One hundred sixty healthy infants at least 12 months old undergoing their first MM immunization in an ambulatory setting. DESIGN AND INTERVENTIONS: Randomized, double-blind, controlled trial of EMLA patch (5%-1 g) or placebo before MMR immunization. Blood sampling before and 28 to 35 days after immunization. OUTCOME MEASURES: The primary outcome measure was the antibody response to measles by plaque reduction neutralization and to mumps and rubella by enzyme immunoassay. The secondary outcomes were pain scores by the Modified Behavioral Pain Scale and drug- and vaccine-associated adverse events. RESULTS: There was no difference in the antibody response between the EMLA- and placebo-treated groups. The response rates in the EMLA group were 89.7%, 88.3%, and 92.3% to measles, mumps, and rubella, respectively, compared with rates of 91.1%, 94.9%, and 93.7% in the placebo group (P >.05 for all comparisons). EMLA recipients had less pain after immunization (mean Modified Behavioral Pain Scale score increase 3.1 compared with 3.8; P =.043) and less irritability (16% compared with 31%; P =.040) than did placebo recipients. CONCLUSIONS: The EMLA patch has no adverse effect on the antibody response to MMR vaccine and significantly reduces the pain associated with the subcutaneous administration of the vaccine. PMID- 10839881 TI - The mother-child interaction and clinical judgment during acute pediatric illnesses. AB - OBJECTIVES: For acutely ill children living in less than optimal environments, mothers and pediatricians may have a heightened perception of illness severity, a lower specificity of clinical judgments, and a tendency to over-utilize resources. We examined the mother-child interaction in order to understand the relation of less optimal environments to clinical judgment and resource use. STUDY DESIGN: At the 2-week and 6-, 15-, and 24-month well child visits of 316 children, the mother-well child interaction was assessed by using the Biringen's Emotional Availability Scales (EAS). Data were gathered regarding maternal depression and sense of competence, infant temperament, maternal social support, life events, the home environment, and demographics. At ill visits, the mother ill child interaction was assessed by using the EAS, and mothers and pediatricians independently assessed illness severity using the Acute Illness Observation Scales. Resource use during the illness was evaluated. RESULTS: One thousand nine hundred eight-three acute illnesses were assessed. A less optimal mother-child interaction was significantly (P <.05 for all comparisons) associated with poorer reliability of mothers' judgments, lower specificity of mothers' judgments (71% vs 85%) and pediatricians' judgments (92% vs 97%), and greater use of resources (eg, for hospitalizations, 2.6% of visits vs 0.7%). Adverse maternal, infant, and demographic characteristics were associated with a less optimal mother-well child (r = 0.68) and mother-ill child (r = 0.80) interaction, a heightened perception of illness severity, and greater resource use. CONCLUSION: Less optimal environments adversely affect the mother-child interaction; a poor mother-child interaction is correlated with low specificity of clinical judgment and over-utilization of resources. PMID- 10839882 TI - Clinicopathologic exercise: hypoglycemia in a young woman with amenorrhea. PMID- 10839883 TI - Accurate diagnosis of Helicobacter pylori infection by stool antigen test and 6 other currently available tests in children. AB - Invasive and noninvasive tests have been developed for the diagnosis of Helicobacter pylori infection. Because H pylori infection is acquired in childhood and adolescence, accurate diagnosis of the infection in the pediatric population is important. We conducted a study to compare invasive tests: culture, biopsy urease test, histology, and polymerase chain reaction on gastric biopsy specimens, with noninvasive tests: serology, (13)C-urea breath test, and a new diagnostic modality, stool antigen test to diagnose H pylori infection. A total of 53 children with symptoms were enrolled in this study, and all had completed the 7 diagnostic tests for H pylori. All the diagnostic tests except serology were excellent methods of diagnosing H pylori infection in children; the diagnostic accuracy was as follows: stool antigen test 96.2%, biopsy urease test 96.2%, histology 98.1%, polymerase chain reaction 94.3%, culture 98.1%, (13)C urea breath test 100%, and serology 84.9%. The stool antigen test, being highly sensitive and specific, will be potentially very helpful in diagnosing H pylori infection in children. PMID- 10839884 TI - Children with ocular motor apraxia type Cogan carry deletions in the gene (NPHP1) for juvenile nephronophthisis. AB - Congenital ocular motor apraxia type Cogan is characterized by impairment of horizontal voluntary eye movements, ocular attraction movements, and optokinetic nystagmus. Two patients with congenital ocular motor apraxia type Cogan exhibited a newly recognized association with nephronophthisis type 1, an autosomal recessive kidney disease. Both patients possess large deletions of the NPHP1 gene. The deletion occurred on both chromosomes 2q13 in one patient and heterozygously in combination with a point mutation of the NPHP1 gene in the other. The findings will help to elucidate the pathogenetic processes involved. PMID- 10839885 TI - Thoracotomy for persistent bronchopleural fistula in the very low birth weight infant. AB - Although conservative treatment is appropriate for most very low birth weight infants with bronchopleural fistulas, early surgical closure may improve survival in properly selected patients. We report our experience with successful surgical closure in 3 consecutive neonates weighing <800 g. PMID- 10839886 TI - Bilateral cystic pleuropulmonary blastoma in early infancy. AB - We report 2 cases of multifocal cystic (type 1) pleuropulmonary blastoma, diagnosed during the first 6 months of life. This rare entity must be recognized before evolution into the prognostically unfavorable type 2 or type 3 pleuropulmonary blastoma. PMID- 10839887 TI - Paravertebral and intraspinal malposition of transfemoral central venous catheters in newborns. AB - We report permanent tetraplegia in a newborn resulting from intraspinal malposition of a transfemoral catheter. In 2 other neonates paravertebral malposition of indwelling Silastic lines was detected. We suggest that left-sided transfemoral catheterization and conditions enhancing collateral flow through the vertebrolumbar pathway may predispose to inadvertent paravertebral catheter placement. PMID- 10839888 TI - Compliance with gluten-free diet in adolescents with screening-detected celiac disease: a 5-year follow-up study. AB - After 5 years of treatment, 22 patients with celiac disease, diagnosed by means of serologic mass screening (mean age, 17.9 years), showed a lower compliance with a gluten-free diet and frequent positivity of serum anti-endomysium antibodies (32%) in comparison with a group of 22 age-matched patients diagnosed because of "typical" symptoms during childhood. PMID- 10839889 TI - Isolated pancreatic amylase deficiency: probable error in maturation. AB - A boy with failure to thrive and isolated pancreatic amylase deficiency is described. Immunoprecipitation confirmed only salivary isoamylase in duodenal fluid at ages 20 and 33 months. Because normal pancreatic amylase messenger RNA was detected by reverse-transcriptase polymerase chain reaction in the fluid, failure of the normal maturation of pancreatic amylase secretion may explain the deficiency. PMID- 10839890 TI - Advances in the diagnosis and management of pediatric seizure disorders in the twentieth century. PMID- 10839891 TI - Optic nerve hypoplasia and congenital hypopituitarism. PMID- 10839892 TI - Adrenal function in preterm infants treated with beclomethasone. PMID- 10839893 TI - Reply PMID- 10839894 TI - Neonatal hypoglycemia: sweet and sour. PMID- 10839895 TI - Reply PMID- 10839896 TI - Red stools and omnicef. PMID- 10839897 TI - Topography of clonidine-induced electroencephalographic changes evaluated by principal component analysis. AB - BACKGROUND: Principal component analysis is a multivariate statistical technique to facilitate the evaluation of complex data dimensions. In this study, principle component analysis was used to reduce the large number of variables from multichannel electroencephalographic recordings to a few components describing changes of spatial brain electric activity after intravenous clonidine. METHODS: Seven healthy volunteers (age, 26 +/- 3 [SD] yr) were included in a double-blind crossover study with intravenous clonidine (1.5 and 3.0 microg/kg). A spontaneous electroencephalogram was recorded by 26 leads and quantified by standard fast Fourier transformation in the delta, theta, alpha, and beta bands. Principle component analysis derived from a correlation matrix calculated between all electroencephalographic leads (26 x 26 leads) separately within each classic frequency band. The basic application level of principle component analysis resulted in components representing clusters of electrodes positions that were differently affected by clonidine. Subjective criteria of drowsiness and anxiety were rated by visual analog scales. RESULTS: Topography of clonidine-induced electroencephalographic changes could be attributed to two independent spatial components in each classic frequency band, explaining at least 85% of total variance. The most prominent effects of clonidine were increases in the delta band over centroparietooiccipital areas and decreases in the alpha band over parietooccipital regions. Clonidine administration resulted in subjective drowsiness. CONCLUSIONS: Data from the current study supported the fact that spatial principle component analysis is a useful multivariate statistical procedure to evaluate significant signal changes from multichannel electroencephalographic recordings and to describe the topography of the effects. The clonidine-related changes seen here were most probably results of its sedative effects. PMID- 10839898 TI - Field evaluation of experimental cardiopulmonary resuscitation techniques. PMID- 10839899 TI - Erythrocyte salvage during cesarean section. PMID- 10839900 TI - Efficacy of continuous insufflation of oxygen combined with active cardiac compression-decompression during out-of-hospital cardiorespiratory arrest. AB - BACKGROUND: During experimental cardiac arrest, continuous insufflation of air or oxygen (CIO) through microcannulas inserted into the inner wall of a modified intubation tube and generating a permanent positive intrathoracic pressure, combined with external cardiac massage, has previously been shown to be as effective as intermittent positive pressure ventilation (IPPV). METHODS: After basic cardiorespiratory resuscitation, the adult patients who experienced nontraumatic, out-of-hospital cardiac arrest with asystole, were randomized to two groups: an IPPV group tracheally intubated with a standard tube and ventilated with standard IPPV and a CIO group for whom a modified tube was inserted, and in which CIO at a flow rate of 15 l/min replaced IPPV (the tube was left open to atmosphere). Both groups underwent active cardiac compression decompression with a device. Resuscitation was continued for a maximum of 30 min. Blood gas analysis was performed as soon as stable spontaneous cardiac activity was restored, and a second blood gas analysis was performed at admission to the hospital. RESULTS: The two groups of patients (47 in the IPPV and 48 in the CIO group) were comparable. The percentages of patients who underwent successful resuscitation (stable cardiac activity; 21.3 in the IPPV group and 27.1% in the CIO group) and the time necessary for successful resuscitation (11.8 +/- 1.8 and 12.8 +/- 1.9 min) were also comparable. The blood gas analysis performed after resuscitation (8 patients in the IPPV and 10 in the CIO group) did not show significant differences. The arterial blood gases performed after admission to the hospital and ventilation using a transport ventilator (seven patients in the IPPV group and six in the CIO group) showed that the partial pressure of arterial carbon dioxide (PaCO2) was significantly lower in the CIO group (35.7 +/- 2.1 compared with 72.7 +/- 7.4 mmHg), whereas the pH and the partial pressure of arterial oxygen (PaO2) were significantly higher (all P < 0.05). CONCLUSIONS: Continuous insufflation of air or oxygen alone through a multichannel open tube was as effective as IPPV during out-of-hospital cardiac arrest. A significantly greater elimination of carbon dioxide and a better level of oxygenation in the group previously treated with CIO probably reflected better lung mechanics. PMID- 10839901 TI - Amniotic fluid removal during cell salvage in the cesarean section patient. AB - BACKGROUND: Cell salvage has been used in obstetrics to a limited degree because of a fear of amniotic fluid embolism. In this study, cell salvage was combined with blood filtration using a leukocyte depletion filter. A comparison of this washed, filtered product was then made with maternal central venous blood. METHODS: The squamous cell concentration, lamellar body count, quantitative bacterial colonization, potassium level, and fetal hemoglobin concentration were measured in four sequential blood samples collected from 15 women undergoing elective cesarean section. The blood samples collected included (1) unwashed blood from the surgical field (prewash), (2) washed blood (postwash), (3) washed and filtered blood (postfiltration), and (4) maternal central venous blood drawn from a femoral catheter at the time of placental separation. RESULTS: Significant reductions in the following parameters were seen when the postfiltration samples were compared to the prewash samples (median [25th-75th percentile]): squamous cell concentration (0.0 [0.0-0.1 counts/high-powered field (HPF)] vs. 8.3 counts/HPF [4. 0-10.5 counts/HPF], P < 0.05); bacterial contamination (0.1 [0.0 0. 2] vs. 3.0 [0.6-7.7] colony-forming units (CFU)/ml, P < 0.01); and lamellar body concentration (0.0 [0.0-1.0] vs. 22.0 [18.5-29.5] thousands/microl, P < 0.01). No significant differences existed between the postfiltration and maternal samples for each of these parameters. Fetal hemoglobin was in higher concentrations in the postfiltration sample when compared with maternal blood (1.9 [1.1-2. 5] vs. 0.5% [0.3-0.7] ). Potassium levels were significantly less in the postfiltration sample when compared with maternal (1.4 [1.0-1.5] vs. 3.8 mEq/l [3.7-4.0]). CONCLUSIONS: Leukocyte depletion filtering of cell-salvaged blood obtained from cesarean section significantly reduces particulate contaminants to a concentration equivalent to maternal venous blood. PMID- 10839902 TI - Intensive care utilization during hospital admission for delivery: prevalence, risk factors, and outcomes in a statewide population. AB - BACKGROUND: During childbirth, the maternal need for intensive care unit (ICU) services is not well-defined. This information could influence the decision whether to incorporate ICU services into the labor and delivery suite. METHODS: This study reports (1) ICU use and mortality rates in a statewide population of obstetric patients during their hospital admission for childbirth, and (2) the risk factors associated with ICU admission and mortality. A case-control design using patient records from a state-maintained anonymous database for the years 1984-1997 was used. Outcome variables included ICU use and mortality rates. RESULTS: Of the 822,591 hospital admissions for delivery of neonates during the study period, there were 1,023 ICU admissions (0.12%) and 34 ICU deaths (3.3%). Age, race, hospital type, volume of deliveries, and source of admission independently and in combination were associated with ICU admission (P < 0.05). The most common risk factors associated with ICU admission included cesarean section, preeclampsia or eclampsia, and postpartum hemorrhage (P < 0.001). Black race, high hospital volume of deliveries, and longer duration of ICU stay were associated with ICU mortality (P < 0.05). The most common risk factors associated with ICU mortality included pulmonary complications, shock, cerebrovascular event, and drug dependence (P < 0.05). CONCLUSIONS: This study shows that ICU use and mortality rate during hospital admission for delivery of a neonate is low. These results may influence the location of perinatal ICU services in the hospital setting. PMID- 10839903 TI - The dose-response of intrathecal sufentanil added to bupivacaine for labor analgesia. AB - BACKGROUND: Regional analgesia for labor often is initiated with an intrathecal injection of a local anesthetic and opioid. The purpose of this prospective, randomized, blinded study was to determine the optimal dose of intrathecal sufentanil when combined with 2.5 mg bupivacaine for labor analgesia. METHODS: One hundred seventy parous parturients with cervical dilation between 3-5 cm were randomized to receive intrathecal 0 (control), 2.5, 5.0, 7.5, or 10.0 microg sufentanil combined with 2.5 mg bupivacaine, followed by a lidocaine epidural test dose, for initiation of analgesia (34 patients in each group). Visual analog scores and the presence of nausea, vomiting, and pruritus were determined every 15 min until the patient requested additional analgesia. Fetal heart rate tracings were compared between groups. RESULTS: Groups were similar for age, height, weight, oxytocin dose, duration of labor, and baseline visual analog scores. Duration of action was significantly shorter for control patients (39 +/- 25 min [mean +/- SD]) compared with those administered sufentanil, all doses (93 +/- 32, 93 +/- 47, 94 +/- 33, 97 +/- 39 min), but was not different among groups administered 2.5, 5.0, 7.5, or 10.0 microg sufentanil. More patients who received 10 microg sufentanil reported nausea and vomiting than did control patients. The severity of pruritus increased with administration of 7.5 and 10.0 microg sufentanil. There was no difference in fetal heart rate changes among groups. CONCLUSIONS: Intrathecal bupivacaine (2.5 mg) without sufentanil did not provide satisfactory analgesia for parous patients. However, bupivacaine combined with 2.5 microg sufentanil provided analgesia comparable to higher doses, with a lower incidence of nausea and vomiting and less severe pruritus. PMID- 10839904 TI - Pharmacokinetics of human cerebral opioid extraction: a comparative study on sufentanil, fentanyl, and alfentanil in a patient after severe head injury. AB - BACKGROUND: The pharmacodynamic differences in time to onset and dissipation of effect of sufentanil, fentanyl, and alfentanil probably result from different rates of blood-brain equilibration. The authors investigated this hypothesis in humans. METHODS: After simultaneous central venous bolus application of sufentanil (10 microg), fentanyl (100 microg), and alfentanil (1,000 microg), arterial and jugular bulb blood samples were drawn simultaneously at 20, 30, 45, 60, 75, 90, 105, 120, 140, 160, 180, 210, 240, 300, 360, and 420 s from 19 patients during the postacute stage of head injury with normal intracranial pressure, cerebral perfusion pressure, and cerebral oxygen metabolism during normocapnia. RESULTS: Peak brain concentration, indicated by equilibrium between arterial and jugular bulb opioid concentrations, was achieved for alfentanil at 45 s, for sufentanil at 5 min, and for fentanyl at 6 min. The corresponding median time intervals (fifth and ninety-fifth percentiles) to reach 50% of peak brain concentration were 15 (14-18), 25 (18-38) and 35 (25-45) s, respectively. Uptake was highest 20 s after bolus and decreased continuously for fentanyl and sufentanil, whereas alfentanil uptake was biphasic. The ratio of the relative amounts of sufentanil, fentanyl, and alfentanil retained in the brain at peak brain concentration was 1x:x6x:x90. CONCLUSIONS: The differences in the time lag between changes in serum concentrations and drug effect after bolus application of nearly equipotent doses of sufentanil, fentanyl, and alfentanil originate from the different times required to reach blood-brain equilibration, mainly depending on different levels and different time profiles of arterial blood concentrations caused by the different tissue distribution volumes. PMID- 10839905 TI - Intraperitoneal and retroperitoneal carbon dioxide insufflation evoke different effects on caval vein pressure gradients in humans: evidence for the starling resistor concept of abdominal venous return. AB - BACKGROUND: The authors hypothesized that intraperitoneal and retroperitoneal carbon dioxide insufflation during surgical procedures evoke markedly different effects on the venous low-pressure system, induce different inferior caval vein pressure gradients at similar insufflation pressures, and may provide evidence for the Starling resistor concept of abdominal venous return. METHODS: Intra- and extrathoracic caval vein pressures were measured using micromanometers during carbon dioxide insufflation at six cavity pressures (baseline and 10, 15, 20, and 24 mmHg and desufflation) in 20 anesthetized patients undergoing laparoscopic (supine, n = 8) or left (n = 6) or right (n = 6) retroperitoneoscopic (prone position) surgery. Intracavital, esophageal, and gastric pressures also were assessed. Data were analyzed for insufflation pressure-dependent and group effects by one-way and two-way analysis of variance for repeated measurements, respectively, followed by the Newman-Keuls post hoc test (P < 0.05). RESULTS: Intraperitoneal, unlike retroperitoneal, insufflation markedly increased, in an insufflation pressure-dependent fashion, the inferior-to-superior caval vein pressure gradient (P < 0.00001) at the level of the diaphragm. In contrast to what was observed with retroperitoneal insufflation, transmural intrathoracic caval vein pressure increased at 10 mmHg insufflation pressure, but the increase flattened with an insufflation pressure of more than 10 mmHg, and pressure decreased with an inflation pressure of 20 mmHg (P = 0.0397). These data are consistent with a zone 2 or 3 abdominal vascular condition during intraperitoneal and a zone 3 abdominal vascular condition during retroperitoneal insufflation. CONCLUSIONS: Intraperitoneal but not retroperitoneal carbon dioxide insufflation evokes a transition of the abdominal venous compartment from a zone 3 to a zone 2 condition, presumably impairing venous return, supporting the Starling resistor concept of abdominal venous return in humans. PMID- 10839906 TI - The influence of drug-induced low plasma cholinesterase activity on the pharmacokinetics and pharmacodynamics of mivacurium. AB - BACKGROUND: The short duration of action of mivacurium results from its rapid hydrolysis by plasma cholinesterase. Bambuterol, an oral bronchodilator, has an inhibiting effect on plasma cholinesterase. The purpose of this study was to evaluate the effect of bambuterol-induced low plasma cholinesterase activity on the pharmacokinetics and pharmacodynamics of mivacurium. METHODS: Fourteen patients received 20 mg bambuterol and 14 patients received placebo orally 2 h before induction of anesthesia. During anesthesia the neuromuscular block was monitored at the thumb using train-of-four nerve stimulation every 12 s and mechanomyography. The times to different levels of neuromuscular recovery after 0.2 mg/kg mivacurium were measured. The concentrations in venous blood of the three isomers and the metabolites of mivacurium were measured using high performance liquid chromatography. RESULTS: Plasma cholinesterase activity was inhibited a median of 90% (range, 67-97%) after bambuterol. The time to first response to train-of-four nerve stimulation was 15 min (range, 9-21 min) and 59 min (range, 32-179 min) in patients receiving placebo and bambuterol, respectively. The estimated clearances of the isomers were significantly lower and the elimination half-lives of all three isomers significantly prolonged in patients receiving bambuterol. No difference was seen in elimination half-lives of the metabolites. The elimination rate constant from the effect compartment and the potency of mivacurium was not affected by bambuterol. CONCLUSION: A 90% inhibition of plasma cholinesterase activity significantly reduced clearance of the isomers of mivacurium. Correspondingly, the duration of action of 0.2 mg/kg mivacurium was prolonged three- to fourfold, compared with patients not administered bambuterol. PMID- 10839907 TI - Ropivacaine, 0.1%, plus sufentanil, 0.5 microg/ml, versus bupivacaine, 0.1%, plus sufentanil, 0.5 microg/ml, using patient-controlled epidural analgesia for labor: a double-blind comparison. AB - BACKGROUND: This study compared the administration of 0.1% ropivacaine and 0.5 microg/ml sufentanil with that of 0.1% bupivacaine and 0.5 microg/ml sufentanil via patient-controlled epidural analgesia route during labor. METHODS: Two hundred healthy pregnant women at term with a single fetus with a vertex fetal presentation were randomized in a double-blind fashion to receive either 0.1% ropivacaine and 0.5 microg/ml sufentanil or 0.1% bupivacaine and 0.5 microg/ml sufentanil using a patient-controlled epidural analgesia pump (5-ml bolus dose, 10-min locked-out period, no basal infusion). Pain score on a visual analog scale, Bromage score (0-3), level of sensory block, patient-controlled epidural analgesia ratio, drug use, supplemental boluses, and side effects were recorded at 30 min and then hourly. Mode of delivery, duration of first and second stages of labor, umbilical cord pH, Apgar scores of the newborn, and a measure of maternal satisfaction were recorded after delivery. RESULTS: No differences were seen between the two groups for pain scores on a visual analog scale during labor, volume of anesthetic solution used, mode of delivery, or side effects. Motor block during the first stage of labor was significantly less in the ropivacaine group than in the bupivacaine group (no motor block in 97.8 of patients vs. 88.3%, respectively; P < 0.01). Duration of the second stage of labor was shorter in the ropivacaine group (1.3 +/- 1.0 vs. 1.5 +/- 1.2 h [mean +/- SD]; P < 0.05). Maternal satisfaction was greater in the bupivacaine group (91 +/- 13 mm for contraction, 89 +/- 19 mm for delivery on a visual scale: 0 = not satisfied at all, 100 = fully satisfied) than in the ropivacaine group (84 +/ 21 and 80 +/- 25 mm; P < 0.0001). Patients in the ropivacaine group requested more supplemental boluses to achieve analgesia during the second stage of labor than those in the bupivacaine group (29.7 vs. 19.8%, respectively, requested one or more supplemental boluses; P < 0.05). CONCLUSIONS: Delivered as patient controlled epidural analgesia, 0.1% ropivacaine and 0.5 microg/ml sufentanil produce less motor block but are clinically less potent than 0.1% bupivacaine and 0.5 microg/ml sufentanil. PMID- 10839909 TI - Response surface model for anesthetic drug interactions. AB - BACKGROUND: Anesthetic drug interactions traditionally have been characterized using isobolographic analysis or multiple logistic regression. Both approaches have significant limitations. The authors propose a model based on response surface methodology. This model can characterize the entire dose-response relation between combinations of anesthetic drugs and is mathematically consistent with models of the concentration-response relation of single drugs. METHODS: The authors defined a parameter, theta, that describes the concentration ratio of two potentially interacting drugs. The classic sigmoid Emax model was extended by making the model parameters dependent on theta. A computer program was used to estimate response surfaces for the hypnotic interaction between midazolam, propofol, and alfentanil, based on previously published data. The predicted time course of effect was simulated after maximally synergistic bolus dose combinations. RESULTS: The parameters of the response surface were identifiable. With the test data, each of the paired combinations showed significant synergy. Computer simulations based on interactions at the effect site predicted that the maximally synergistic three-drug combination tripled the duration of effect compared with propofol alone. CONCLUSIONS: Response surfaces can describe anesthetic interactions, even those between agonists, partial agonists, competitive antagonists, and inverse agonists. Application of response surface methodology permits characterization of the full concentration-response relation and therefore can be used to develop practical guidelines for optimal drug dosing. PMID- 10839908 TI - Reliability of the heparin management test for monitoring high levels of unfractionated heparin: in vitro findings in volunteers versus in vivo findings during cardiopulmonary bypass. AB - BACKGROUND: The authors assessed the heparin management test in vitro in volunteers and in vivo during cardiopulmonary bypass. METHODS: In vitro, the heparin management test was analyzed for heparin levels between 0 and 6 IU/ml using variations in hematocrit, platelets, procoagulants, and storage time. The in vivostudies consisted of two groups: In group I (cardiopulmonary bypass /= 180 min, with aprotinin) included use (n = 10) and nonuse of coumadin (n = 10) and anticoagulation according to the automated heparin dose-response assay. Tests were performed in duplicate (whole blood, two heparin management test analyzers) and compared with anti-Xa activity (plasma). RESULTS: In vitro, the results of the heparin management test (n = 1,070) correlated well with heparin concentration (r2 = 0.98). Dilution and storage time did not affect the heparin management test; a hematocrit of 60% and reduced procoagulants (10%) prolonged clotting time. In vivo, the correlation (heparin management test vs. anti-Xa) was strong in group I (r2 = 0.97 [with aprotinin] and 0.96 [without aprotinin]; n = 960) and group II without coumadin (r2 = 0.89, n = 516). In group II with coumadin, the overall correlation was r2 = 0.87 and 0.79 (n = 484), although the range varied widely (0.57-0.94, between-analyzer differences 0-47%). CONCLUSIONS: The results of the heparin management test were influenced by hematocrit, plasma coagulation factors, and the heparin level, but not by use of aprotinin. The heparin management test provided reliable values in vitro in group I, and in group II without coumadin but was less reliable in group II with coumadin. PMID- 10839910 TI - Multiport epidural catheters: does the air test work? AB - BACKGROUND: Multiport epidural catheters are popular; however, the reliability of the air test has not been evaluated with this catheter design. The authors determined the effectiveness of aspirating for blood and the air test in detecting intravascular multiorifice epidural catheters. METHODS: Three hundred women in labor underwent placement of a blunt-tip, three-hole, 20-gauge, lumbar epidural catheter. If there were no signs of spinal anesthesia, 3 ml lidocaine or bupivacaine was injected and the patient was observed for signs of spinal anesthesia. If there were no signs of spinal anesthesia, the authors injected 1 ml air through the epidural catheter while listening to the maternal precordium using a Doppler fetal heart rate monitor. Catheters through which blood was aspirated were air-tested and replaced. Patients with air-test-positive, blood aspiration-negative catheters received 100 mg lidocaine through the catheter and were questioned about toxicity symptoms. The authors injected bupivacaine fentanyl through aspiration-negative,air-test-negative catheters and recorded the sensory analgesic level 20 min later. RESULTS: The authors aspirated cerebrospinal fluid through one catheter and documented intravascular placement in 11 catheters. Results of the air test and blood aspiration were positive for eight catheters. Blood could not be aspirated from one air-test-positive catheter; perioral numbness developed in the patient after lidocaine injection. Blood was freely aspirated from two air-test-negative catheters. In the remaining 288 catheters, bupivacaine-fentanyl injection produced epidural analgesia in 279 patients and no effect in 9 patients. CONCLUSIONS: The authors obtained false negative results with both catheter aspiration and the air test. Fractionating the local anesthetic dose is important when using multiorifice epidural catheters. PMID- 10839911 TI - Pressure support ventilation versus continuous positive airway pressure with the laryngeal mask airway: a randomized crossover study of anesthetized adult patients. AB - BACKGROUND: The authors tested the hypothesis that pressure support ventilation (PSV) provides more effective gas exchange than does unassisted ventilation with continuous positive airway pressure (CPAP) in anesthetized adult patients treated using the laryngeal mask airway. METHODS: Forty patients were randomized to two equal-sized crossover groups, and data were collected before surgery. In group 1, patients underwent CPAP, PSV, and CPAP in sequence. In group 2, patients underwent PSV, CPAP, and PSV in sequence. PSV comprised positive end expiratory pressure set at 5 cm H2O and inspiratory pressure support set at 5 cm H2O above positive end expiratory pressure. CPAP was set at 5 cm H2O. Each ventilatory mode was maintained for 10 min. The following data were recorded every minute for the last 5 min of each ventilatory mode and the average reading taken: end tidal carbon dioxide, oxygen saturation, expired tidal volume, leak fraction, respiratory rate, noninvasive mean arterial pressure, and heart rate. RESULTS: In both groups, PSV showed lower end tidal carbon dioxide (P < 0.001), higher oxygen saturation, (P < 0.001), and higher expired tidal volume (P < 0.001) compared with CPAP. In both groups, PSV had similar leak fraction, respiratory rate, mean arterial pressure, and heart rate compared with CPAP. In group 1, measurements for CPAP were similar before and after PSV. In group 2, measurements for PSV were similar before and after CPAP. CONCLUSION: The authors concluded that PSV provides more effective gas exchange than does unassisted ventilation with CPAP during LMA anesthesia while preserving leak fraction and hemodynamic homeostasis. PMID- 10839912 TI - Preemptive analgesia by intravenous low-dose ketamine and epidural morphine in gastrectomy: a randomized double-blind study. AB - BACKGROUND: Morphine and ketamine may prevent central sensitization during surgery and result in preemptive analgesia. The reliability of preemptive analgesia, however, is controversial. METHODS: Gastrectomy patients were given preemptive analgesia consisting of epidural morphine, intravenous low-dose ketamine, and combinations of these in a randomized, double-blind manner. Postsurgical pain intensity was rated by a visual analog scale, a categoric pain evaluation, and cumulative morphine consumption. RESULTS: Preemptive analgesia by epidural morphine and by intravenous low-dose ketamine were significantly effective but not definitive. With epidural morphine, a significant reduction in visual analog scale scores at rest was observed at 24 and 48 h, and morphine consumption was significantly lower at 6 and 12 h, compared with control values. With intravenous ketamine, visual analog scale scores at rest and morphine consumption were significantly lower at 6, 12, 24, and 48 h than those in control subjects. The combination of epidural morphine and intravenous ketamine provided definitive preemptive analgesia: Visual analog scale scores at rest and morphine consumption were significantly the lowest at 6, 12, 24, and 48 h, and the visual analog scale score during movement and the categoric pain score also were significantly the lowest among the groups. CONCLUSION: The results suggest that for definitive preemptive analgesia, blockade of opioid and N-methyl-d-aspartate receptors is necessary for upper abdominal surgery such as gastrectomy; singly, either treatment provided significant, but not definitive, postsurgical pain relief. Epidural morphine may affect the spinal cord segmentally, whereas intravenous ketamine may block brain stem sensitization via the vagus nerve during upper abdominal surgery. PMID- 10839913 TI - The fiberscopic findings of the epidural space in pregnant women. AB - BACKGROUND: The spread of epidural analgesia is facilitated by pregnancy. Changes in the epidural structure during pregnancy may affect the spread of analgesia in pregnant women. To investigate the changes in the epidural space produced by pregnancy, the authors performed epiduroscopy in pregnant women. METHODS: Using a flexible fiberscope, the authors evaluated the epidural space in 73 women undergoing lumbar epidural anesthesia. Patients were classified into three groups: a nonpregnant group (n = 21), a first trimester pregnant group (8-13 weeks, n = 23), and a third-trimester pregnant group (27-39 weeks, n = 29). A 17 gauge Tuohy needle was inserted using the paramedian technique and the loss-of resistance method with 5 ml air. The epiduroscope was introduced into the lumbar epidural space via the Tuohy needle and was advanced approximately 10 cm in a cephalad direction from the needle tip within the epidural space. The differences in the epidural space among the three groups then was evaluated. RESULTS: The epiduroscopy showed that the epidural pneumatic space, after injection of a given amount of air, was narrower and the density of the vascular network greater in the third-trimester group than in the other two groups. The amount of engorged blood vessels was greater in the third and first trimester groups than in the nonpregnant group. The amount of bleeding at the needle tip and the amount of fatty and fibrous connective tissue did not differ among the three groups. CONCLUSIONS: Epidural blood vessels become engorged in the first trimester; the density of the vascular networks increase in the third trimester. These changes in the epidural space during pregnancy may affect the spread of epidural analgesia in pregnant women. PMID- 10839914 TI - Port-access minimally invasive cardiac surgery increases surgical complexity, increases operating room time, and facilitates early postoperative hospital discharge. AB - BACKGROUND: Proposed advantages of port-access cardiac surgery have yet to be substantiated. The authors retrospectively compared patients undergoing port access cardiac surgery with a matched group undergoing conventional cardiac surgery. METHODS: Forty-six patients who underwent port-access cardiac surgery were matched with 46 who underwent conventional cardiac surgery. Absolute criteria for matching included morning-of-surgery admission, procedure undergone, and care being delivered by one of two surgeons. If possible, matching included care delivered by one of two anesthesiologists. Patients were matched as closely as possible for preoperative demographic and clinical characteristics. RESULTS: All 46 pairs of patients were matched for procedure and admitted the morning of surgery. All 92 operations were performed by one of two surgeons, and 89% were performed by one of two anesthesiologists. Preoperative demographic and clinical characteristics were equivalent between groups. Compared with conventional cardiac surgery, port-access cardiac surgery increased surgical complexity (it almost tripled cardiopulmonary bypass time during coronary artery bypass grafting and increased it almost 40% during mitral valve procedures) and increased total operating room time (P < 0.0001). Port-access cardiac surgery had no beneficial effect on earlier postoperative extubation, decreased incidence of atrial fibrillation, or intensive care unit time, yet it decreased postoperative duration of stay (P = 0.029, all patients), a benefit observed primarily in patients undergoing coronary artery bypass grafting (P = 0.002). CONCLUSIONS: This retrospective analysis revealed that port-access cardiac surgery increases surgical complexity, increases operating room time, has no effect on earlier postoperative extubation or decreased incidence of atrial fibrillation or intensive care unit time, and may facilitate postoperative hospital discharge (primarily in patients undergoing coronary artery bypass grafting). Properly designed prospective investigation is necessary to ascertain whether port-access cardiac surgery truly offers any benefits over conventional cardiac surgery. PMID- 10839915 TI - Acute severe isovolemic anemia impairs cognitive function and memory in humans. AB - BACKGROUND: Erythrocytes are transfused to prevent or treat inadequate oxygen delivery resulting from insufficient hemoglobin concentration. Previous studies failed to find evidence of inadequate systemic oxygen delivery at a hemoglobin concentration of 5 g/dl. However, in those studies, sensitive, specific measures of critical organ function were not used. This study tested the hypothesis that acute severe decreases of hemoglobin concentration alters human cognitive function. METHODS: Nine healthy volunteers, age 29 +/- 5 yr (mean +/- SD), were tested with verbal memory and standard, computerized neuropsychologic tests before and after acute isovolemic reduction of their hemoglobin to 7, 6, and 5 g/dl and again after transfusion of their autologous erythrocytes to return their hemoglobin concentration to 7 g/dl. To control for duration of the experiment, each volunteer also completed the same tests on a separate day, without alteration of hemoglobin, at times of the day approximately equivalent to those on the experimental day. RESULTS: No test showed any change in reaction time or error rate at hemoglobin concentration of 7 g/dl compared with the data at the baseline hemoglobin concentration of 14 g/dl. Reaction time, but not error rate, for horizontal addition and digit-symbol substitution test (DSST) increased at hemoglobin 6 g/dl (mean horizontal addition, 19%; 95% confidence interval [CI], 4 34%; mean DSST, 10%; 95% CI, 4-17%) and further at 5 g/dl (mean horizontal addition, 43%; 95% CI, 6-79%; mean DSST, 18%; 95% CI, 4-31%). Immediate and delayed memory was degraded at hemoglobin 5 g/dl but not at 6 g/dl. Return of hemoglobin to 7 g/dl returned all tests to baseline, except for the DSST, which significantly improved, and returned to baseline the following morning after transfusion of all autologous erythrocytes. CONCLUSION: Acute reduction of hemoglobin concentration to 7 g/dl does not produce detectable changes in human cognitive function. Further reduction of hemoglobin level to 6 and 5 g/dl produces subtle, reversible increases in reaction time and impaired immediate and delayed memory. These are the first prospective data to demonstrate subtle degraded human function with acute anemia of hemoglobin concentrations of 6 and 5 g/dl. This reversibility of these decrements with erythrocyte transfusion suggests that our model can be used to test the efficacy of erythrocytes, oxygen therapeutics, or other treatments for acute anemia. PMID- 10839916 TI - Pain relief in complex regional pain syndrome due to spinal cord stimulation does not depend on vasodilation. AB - BACKGROUND: Spinal cord stimulation (SCS) is known to relieve pain in patients with complex regional pain syndrome (CRPS) and, in general, to cause vasodilation. The vasodilatory effect of SCS is hypothesized to be secondary to inhibition of sympathetically mediated vasoconstriction, or through antidromic impulses resulting in release of vasoactive substances. The aim of the present study was to assess whether pain relief in CRPS after SCS is, in fact, dependent on vasodilation. In addition, we tried to determine which of the potential mechanisms may cause the vasodilatory effect that is generally found after SCS. METHODS: Twenty-four of 36 patients with unilateral CRPS responded to the test of SCS. Twenty-two of these 24 responders (hand, n = 14; foot, n = 8) who had undergone previous sympathectomy were enrolled for the study. In addition, 20 control subjects (10 controls for each extremity) were studied. By means of laser Doppler flowmetry, the skin microcirculation of the patients was measured bilaterally while the SCS system was switched off and while it was activated. Control subjects (n = 20) were tested once only. The ratio of the rest flow at heart level and the dependent position was defined as the vasoconstriction index. RESULTS: Both in affected hands and feet, patients were found to have lower vasoconstriction indices (P < 0.01) as compared with controls, indicating a decreased sympathetic tone. Applying SCS did not result in any microcirculatory change as compared with baseline or the contralateral clinically unaffected side. CONCLUSIONS: The current study failed to show that SCS influences skin microcirculation in patients with CRPS and a low sympathetic tone. Therefore, we may conclude that pain relief in CRPS due to SCS is possible without vasodilation. Because sympathetic activity was greatly decreased in our patients, these results support the hypothesis that the vasodilation that is normally found with SCS is due to an inhibitory effect on sympathetically maintained vasoconstriction. PMID- 10839917 TI - Area under the plasma concentration-time curve of inorganic fluoride following sevoflurane anesthesia correlates with CYP2E1 mRNA level in mononuclear cells. AB - BACKGROUND: Because the amount of inorganic fluoride released after anesthesia with sevoflurane depends on the dose of administered sevoflurane and cytochrome P450 (CYP) 2E1 activity in the liver, a reliable and noninvasive probe for CYP2E1 would be useful for predicting plasma inorganic fluoride levels after anesthesia. In this study, the authors evaluated the relation between plasma concentration of inorganic fluoride after sevoflurane anesthesia and CYP2E1 mRNA level in mononuclear cells. METHODS: Twenty patients (American Society of Anesthesiologists physical status I), aged 20-68 yr undergoing body surface surgery with general anesthesia with sevoflurane were enrolled. One milliliter of blood was obtained before administration of sevoflurane and mononuclear cells were obtained. Levels of CYP2E1 mRNA in mononuclear cells were measured by competitive reverse transcription polymerase chain reaction with a specific primer and competitor for CYP2E1 mRNA. RESULTS: There was a significant correlation between level of CYP2E1 mRNA in mononuclear cells and the area under the plasma concentration-time curve of plasma inorganic fluoride from the beginning of sevoflurane administration to infinity in uninduced and uninhibited patients (r2 = 0.56; P < 0.01). CONCLUSIONS: Area under the plasma concentration time curve of inorganic fluoride after sevoflurane anesthesia correlates with CYP2E1 mRNA in mononuclear cells in peripheral blood. PMID- 10839918 TI - Differential block of fast and slow inactivating tetrodotoxin-sensitive sodium channels by droperidol in spinal dorsal horn neurons. AB - BACKGROUND: Dorsal horn neurons of the spinal cord participate in neuronal pain transmission. During spinal and epidural anesthesia, dorsal horn neurons are exposed to local anesthetics and opioids. Droperidol is usually given with opioids to avoid nausea and vomiting. A recently developed method of "entire soma isolation" has made it possible to study directly the action of droperidol on different components of Na+ current in dorsal horn neurons. METHODS: Using a combination of the whole-cell patch-clamp recording from spinal cord slices and the entire soma isolation method, we studied the direct action of droperidol on two types of Na+ currents in dorsal horn neurons of young rats. RESULTS: The tetrodotoxin-sensitive Na+ current in isolated somata consisted of a fast inactivating (tauF, 0.5-2 ms; 80-90% of the total amplitude) and a slow inactivating (tauS, 6-20 ms; 10-20% of the total amplitude) component. Droperidol, at concentrations relevant for spinal and epidural anesthesia, selectively and reversibly suppressed the fast component with a half-maximum inhibiting concentration (IC50) of 8.3 microm. The slow inactivating component was much less sensitive to droperidol; the estimated IC50 value was 809 microm. CONCLUSIONS: Droperidol selectively blocks fast Na+ channels, the fast and slow components of the Na+ current in dorsal horn neurons are carried through pharmacologically distinct types of Na+ channels, and the effects of droperidol differ from those of local anesthetics and tetrodotoxin, which equipotently suppress both components. Droperidol may be suggested as a pharmacologic tool for separation of different types of inactivating tetrodotoxin-sensitive Na+ channel. PMID- 10839919 TI - Morphine inhibits NF-kappaB nuclear binding in human neutrophils and monocytes by a nitric oxide-dependent mechanism. AB - BACKGROUND: The transcription factor NF-kappaB plays a pivotal role in gene expression of inflammatory mediators such as cytokines or adhesion molecules. NF kappaB-mediated transcriptional activation of these genes is inhibited by nitric oxide (NO) in a variety of cells, including monocytes. Morphine mediates NO release in a naloxone antagonizable manner in monocytes and neutrophils. METHODS: The influence of morphine on NF-kappaB activation was investigated in a whole blood flow cytometric assay. A specific antibody against the p65 subunit of NF kappaB was used and detected by fluoresceine-isothiocyanate-labeled anti immunoglobulin G. Nuclei were stained with propidium iodide. Leukocyte subpopulations were evaluated by gating on neutrophils and monocytes. The median fluorescence channel was determined. Different morphine concentrations (50 nm, 50 microm, 1 mm) and incubation intervals (10-150 min) were used. RESULTS: Morphine inhibits lipopolysaccharide-induced NF-kappaB nuclear binding in human blood neutrophils and monocytes in a time-, concentration-, and naloxone-sensitive dependent manner. Similar effects were achieved with the NO donor S-nitroso-N acetyl-pencillamine and the antioxidant N-acetyl-cysteine. The NO synthase inhibitors Nomega-nitro-l-arginine-methyl-esther and Nomega-nitro-l-arginine completely abolished the morphine-induced attenuation of NF-kappaB nuclear binding, demonstrating that the inhibitory action is mediated by NO release. CONCLUSION: Morphine causes immunosuppression, at least in part, via the NO stimulated depression of NF-kappaB nuclear binding. PMID- 10839920 TI - Intravenous lidocaine inhibits visceral nociceptive reflexes and spinal neurons in the rat. AB - BACKGROUND: Systemically administered local anesthetics and other sodium channel blockers produce analgesia in patients with hypersensitivity disorders. To assess whether these agents have a role in the treatment of visceral pain, the present study examined the effects of intravenous lidocaine on neuronal and reflex responses to colorectal distension. METHODS: In decerebrate, cervical spinal cord transected male rats, the lumbosacral spinal cord was exposed by a laminectomy. Dorsal horn neurons demonstrating excitatory responses to colorectal distension were identified using microelectrodes. Sequential doses of lidocaine were administered intravenously. In chronically instrumented, unanesthetized rats, visceromotor responses, pressor responses, and increases in heart rate were elicited by colorectal distension and sequential doses of lidocaine. RESULTS: Intravenous lidocaine dose-dependently inhibited visceromotor and cardiovascular reflexes and the evoked and spontaneous activity of neurons excited by colorectal distension. There were statistically greater effects on one of the neuronal subgroups (sustained neurons) than on another subgroup (abrupt neurons.) CONCLUSIONS: Intravenous lidocaine had dose-dependent, inhibitory effects on two spinal neuronal populations excited by colorectal distension and dose-dependently inhibited reflex responses to the same stimulus. This suggests there may be utility of sodium channel blockers in the treatment of pain of visceral origin. PMID- 10839921 TI - Isoflurane, but not halothane, induces protection of human myocardium via adenosine A1 receptors and adenosine triphosphate-sensitive potassium channels. AB - BACKGROUND: Volatile anesthetics produce differing degrees of myocardial protection in animal models of ischemia. The purpose of the current investigation was to determine the influence of isoflurane and halothane on myocardial protection in a human model of simulated ischemia and the role of adenosine A1 receptors and adenosine triphosphate-sensitive potassium (KATP) channels in the anesthetic pathway. METHODS: Human atrial trabecular muscles were superfused with oxygenated Krebs-Henseleit buffer and stimulated at 1 Hz, with recording of maximum contractile force. Fifteen minutes before a 30-min anoxic insult, muscles were pretreated for 5 min with either anoxia, the A1 agonist N6 cyclohexyladenosine, 1% halothane or 1.2% isoflurane. These treatments were also performed in the presence of either the KATP channel antagonist glibenclamide or the A1 receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX). Anesthetic effects were also determined on KATP currents in isolated whole cell voltage clamped human atrial myocytes. RESULTS: Recovery of force (recorded 60 min after anoxia) in isoflurane-pretreated muscles was reduced from 76.6 +/- 7.5% of baseline to 43.7 +/- 7.1% by pretreatment with glibenclamide, and to 52.5 +/- 6.2% by pretreatment with DPCPX. Halothane treatment provided no cardioprotection and seemed to inhibit protection by anoxic preconditioning. Halothane decreased whole cell KATP currents in atrial myocytes, whereas isoflurane had no effects. CONCLUSIONS: This study demonstrates the cardioprotective effects of isoflurane in contrast to the effects of halothane. Furthermore, A1 receptors and KATP channels seem to mediate the beneficial effects of anoxia and isoflurane in human myocardium. PMID- 10839922 TI - Sildenafil is a pulmonary vasodilator in awake lambs with acute pulmonary hypertension. AB - BACKGROUND: Phosphodiesterase type 5 (PDE5) hydrolyzes cyclic guanosine monophosphate in the lung, thereby modulating nitric oxide (NO)/cyclic guanosine monophosphate-mediated pulmonary vasodilation. Inhibitors of PDE5 have been proposed for the treatment of pulmonary hypertension. In this study, we examined the pulmonary and systemic vasodilator properties of sildenafil, a novel selective PDE5 inhibitor, which has been approved for the treatment of erectile dysfunction. METHODS: In an awake lamb model of acute pulmonary hypertension induced by an intravenous infusion of the thromboxane analog U46619, we measured the effects of 12.5, 25, and 50 mg sildenafil administered via a nasogastric tube on pulmonary and systemic hemodynamics (n = 5). We also compared the effects of sildenafil (n = 7) and zaprinast (n = 5), a second PDE5 inhibitor, on the pulmonary vasodilator effects of 2.5, 10, and 40 parts per million inhaled NO. Finally, we examined the effect of infusing intravenous l-NAME (an inhibitor of endogenous NO production) on pulmonary vasodilation induced by 50 mg sildenafil (n = 6). RESULTS: Cumulative doses of sildenafil (12.5, 25, and 50 mg) decreased the pulmonary artery pressure 21%, 28%, and 42%, respectively, and the pulmonary vascular resistance 19%, 23%, and 45%, respectively. Systemic arterial pressure decreased 12% only after the maximum cumulative sildenafil dose. Neither sildenafil nor zaprinast augmented the ability of inhaled NO to dilate the pulmonary vasculature. Zaprinast, but not sildenafil, markedly prolonged the duration of pulmonary vasodilation after NO inhalation was discontinued. Infusion of l-NAME abolished sildenafil-induced pulmonary vasodilation. CONCLUSIONS: Sildenafil is a selective pulmonary vasodilator in an ovine model of acute pulmonary hypertension. Sildenafil induces pulmonary vasodilation via a NO dependent mechanism. In contrast to zaprinast, sildenafil did not prolong the pulmonary vasodilator action of inhaled NO. PMID- 10839923 TI - Multiple agents potentiate alpha1-adrenoceptor-induced conduction depression in canine cardiac purkinje fibers. AB - BACKGROUND: Halothane more so than isoflurane potentiates an alpha1-adrenoceptor (alpha1-AR)-mediated action of epinephrine that abnormally slows conduction in Purkinje fibers and may facilitate reentrant arrhythmias. This adverse drug interaction was further evaluated by examining conduction responses to epinephrine in combination with thiopental and propofol, which "sensitize" or reduce the dose of epinephrine required to induce arrhythmias in the heart, and with etomidate, which does not, and responses to epinephrine with verapamil, lidocaine, and l-palmitoyl carnitine, a potential ischemic metabolite. METHODS: Action potentials and conduction times were measured in vitro using two microelectrodes in groups of canine Purkinje fibers stimulated at 150 pulses/min. Conduction was evaluated each minute after exposure to 5 microm epinephrine (or phenylephrine) alone or with the test drugs. Changes in the rate of phase 0 depolarization (Vmax) and the electrotonic spread of intracellular current were measured during exposure to epinephrine with octanol to evaluate the role of inhibition of active and passive (intercellular coupling) membrane properties in the transient depression of conduction velocity. RESULTS: Lidocaine (20 microm) and octanol (0.2 mm) potentiated alpha1-AR-induced conduction depression like halothane (0.4 mm), with maximum depression at 3-5 min of agonist exposure, no decrease of Vmax, and little accentuation at a rapid (250 vs. 150 pulses/min) stimulation rate. Thiopental (95 microm), propofol (50 microm), and verapamil (2 microm) similarly potentiated epinephrine responses, whereas etomidate (10 microm) did not. Between groups, the decrease of velocity induced by epinephrine in the presence of (10 microm) l-palmitoyl carnitine (-18%) was significantly greater than that resulting from epinephrine alone (-6%; 0.05 10 and <100 cfu/ml. Eucaryotic DNA rich normal mouse lung homogenate did not interfere among these samples. The method amplified the 216 bp product also among cfu positive tissues of naturally infected mice. Finally, in a mouse model of dormancy, direct RT-PCR detected a signal among multiple tissues that were negative for cfus and hence non culturable. Ag85B is abundantly secreted by MTB and hyper-expressed under stress conditions. Thus the method to identify its mRNA message may be useful to detect viable but dormant bacteria. PMID- 10839971 TI - Early events and implication of F-actin and annexin I associated structures in the phagocytic uptake of Brucella suis by the J-774A.1 murine cell line and human monocytes. AB - Brucella spp. are facultative, intracellular pathogenic bacteria that cause brucellosis, a zoonosis affecting mammalian species. Brucella entry into myelomonocytic cell lines is highly enhanced by opsonization. Few studies have been undertaken to unravel the first interactions between these bacteria and their host cells. This paper deals with early events following contact of Brucella suis with the J-774A.1 phagocytic cell line and differentiated monocytes. Phagocytic uptake of bacteria was documented under a fluorescence microscope using GFP-expressing B. suis. Unlike entry in the J-774A. 1 cell line, non-opsonized Brucella entered differentiated human monocytes as efficiently as opsonized bacteria. However, following 1 h infections, a mean of only three bacteria were phagocytized and the whole monocyte population was only infected after a 4 h infection. Contact of non-opsonized Brucella with phagocytes did not induce marked structural changes at the cell surface, as revealed by scanning electron microscopy. Contact of Brucella (opsonized or not) elicited transient local recruitment of F-actin, revealed by phalloidin labelling, and of annexin I associated structures, revealed by immunofluorescence staining. Finally, bacteria appeared to be rapidly internalized in monocytes once they had adhered to the cell surface. A low percentage of infected cells and few adhered and/or internalized bacteria following short-term infections could have resulted either from the fact that there were few sites of entry or the weak bacterial initial interactions with the host-cell membrane or the bacterial receptor. PMID- 10839972 TI - Protease-resistant prion protein in brain and lymphoid organs of sheep within a naturally scrapie-infected flock. AB - The hallmark of transmissible spongiform encephalopathies (TSE), such as scrapie in sheep, is the accumulation in tissues of an insoluble and protease resistant form (PrPres) of the cellular prion protein. In this study, we evaluated whether the diversity in both the clinical pattern and the PrP genotypes of scrapied sheep from the same flock was connected with different levels and/or glycoform patterns of the PrPres in the brain and lymphoid organs of the animals. Whereas the PrPres levels in spleen, lymph nodes and tonsils from sheep of different PrP genotypes and clinical status appeared comparable, they were highly variable in brain, particularly in the brain stem and the cerebellum. PrPres was only detected in sheep bearing at least one VRQ allele, including three asymptomatic sheep and the highest PrPres load was found in the cerebellum of VRQ/VRQ animals. All together, levels of PrPres in brain did not necessarily correlate with the severity of the clinical disease but might depend on the PrP genotype of the animals. Different brain regions from a given sheep displayed a similar glycopattern of PrPres, whereas the apparent molecular sizes of the unglycosylated and diglycosylated forms of the protein differed between brain and lymphoid tissues. We did not find any notifiable differences in the glycopattern of PrPres in brain from sheep of different PrP genotypes or different clinical status and this PrPres glycotype was also similar to that found in brain from four cattle BSE. PMID- 10839973 TI - Characterization of surface layer proteins from different Clostridium difficile clinical isolates. AB - In a previous study we suggested that two surface proteins of a Clostridium difficile strain were involved in the formation of a regularly assembled surface layer (S-layer) external to the cell wall. In the present paper six C. difficile strains isolated from cases and healthy carriers were studied. By using freeze etching and negative staining techniques two superimposed structurally different lattices were detected on the cell surface of the different C. difficile strains. In each strain, the outer S-layer lattice was arranged in a square symmetry and the inner S-layer lattice in hexagonal symmetry. The S-layer proteins from the different strains were isolated and characterized. Each strain showed two distinct S-layer glycoproteins ranging in molecular mass 36-56 kDa. Antigenic cross-reactivity among the S-layer proteins of higher molecular masses extracted from each strain was demonstrated whereas no antigenic relationship was observed among the different S-layer proteins of lower molecular masses. N-terminal sequence analysis showed the presence of common structural motifs conserved among the high S-layer proteins as well as among the low S-layer proteins. These data indicate that the presence of S-layer on C. difficile strains is common and that its glycoprotein subunits show a certain degree of heterogeneity. PMID- 10839975 TI - Volume contents and index PMID- 10839974 TI - Evaluation of invasion-conferring genotypes and antibiotic-induced hyperinvasive phenotypes in multiple antibiotic resistant Salmonella typhimurium DT104. AB - Antibiotic resistance in pathogenic bacteria is a problem in both industrialized and developing countries. This is especially evident in Salmonella typhimurium, a foodborne pathogen that causes gastrointestinal and systemic disease throughout the world. S. typhimurium DT104 further poses a major health concern due to its apparent enhanced ability to acquire multiple antibiotic resistance genes and its putative hypervirulent phenotype. Recently, we demonstrated that multiresistant S. typhimurium do not appear to be more invasive than non-resistant cohorts. In the present study, we evaluated the presence of Salmonella pathogenicity island 1 (SPI1) flanking and internal sequences in over 400 isolates of multiresistant S. typhimurium. With these same isolates, we also used a tissue culture invasion assay to evaluate a potential relationship between antibiotic exposure and a hyperinvasive phenotype. Our studies revealed that SPI1 flanking sequences are similar in multiresistant and non-resistant S. typhimurium. Furthermore, we failed to identify any isolates that were hyperinvasive in the presence of any of the 14 antibiotics evaluated. These results further indicate that the putative hypervirulence of multiresistant S. typhimurium is not likely to occur at the level of invasion. PMID- 10839976 TI - Novel HOXA13 mutations and the phenotypic spectrum of hand-foot-genital syndrome. AB - Hand-foot-genital syndrome (HFGS) is a rare, dominantly inherited condition affecting the distal limbs and genitourinary tract. A nonsense mutation in the homeobox of HOXA13 has been identified in one affected family, making HFGS the second human syndrome shown to be caused by a HOX gene mutation. We have therefore examined HOXA13 in two new and four previously reported families with features of HFGS. In families 1, 2, and 3, nonsense mutations truncating the encoded protein N-terminal to or within the homeodomain produce typical limb and genitourinary abnormalities; in family 4, an expansion of an N-terminal polyalanine tract produces a similar phenotype; in family 5, a missense mutation, which alters an invariant domain, produces an exceptionally severe limb phenotype; and in family 6, in which limb abnormalities were atypical, no HOXA13 mutation could be detected. Mutations in HOXA13 can therefore cause more-severe limb abnormalities than previously suspected and may act by more than one mechanism. PMID- 10839977 TI - Split-hand/split-foot malformation is caused by mutations in the p63 gene on 3q27. AB - Split-hand/split-foot malformation (SHFM), a limb malformation involving the central rays of the autopod and presenting with syndactyly, median clefts of the hands and feet, and aplasia and/or hypoplasia of the phalanges, metacarpals, and metatarsals, is phenotypically analogous to the naturally occurring murine Dactylaplasia mutant (Dac). Results of recent studies have shown that, in heterozygous Dac embryos, the central segment of the apical ectodermal ridge (AER) degenerates, leaving the anterior and posterior segments intact; this finding suggests that localized failure of ridge maintenance activity is the fundamental developmental defect in Dac and, by inference, in SHFM. Results of gene-targeting studies have demonstrated that p63, a homologue of the cell-cycle regulator TP53, plays a critically important role in regulation of the formation and differentiation of the AER. Two missense mutations, 724A-->G, which predicts amino acid substitution K194E, and 982T-->C, which predicts amino acid substitution R280C, were identified in exons 5 and 7, respectively, of the p63 gene in two families with SHFM. Two additional mutations (279R-->H and 304R-->Q) were identified in families with EEC (ectrodactyly, ectodermal dysplasia, and facial cleft) syndrome. All four mutations are found in exons that fall within the DNA-binding domain of p63. The two amino acids mutated in the families with SHFM appear to be primarily involved in maintenance of the overall structure of the domain, in contrast to the p63 mutations responsible for EEC syndrome, which reside in amino acid residues that directly interact with the DNA. PMID- 10839979 TI - Auto-catalysed progression of aneuploidy explains the Hayflick limit of cultured cells, carcinogen-induced tumours in mice, and the age distribution of human cancer. AB - Evidence continues to accumulate that aneuploidy, an imbalance in the number of chromosomes, is responsible for the characteristic phenotypes of cancer, including the abnormal cellular size and morphology of cancer cells, the appearance of tumour-associated antigens, as well as the high levels of membrane bound and secreted proteins responsible for invasiveness and loss of contact inhibition. Aneuploidy has also been demonstrated to be the self-perpetuating source of the karyotypic instability of cancer cells. Here it is shown that the auto-catalysed progression of aneuploidy explains the kinetics of the finite lifetime of diploid cells in culture, the time course of the appearance of papillomas and carcinomas in benzo[a]pyrene-treated mice, and the age-dependence of human cancers. Modelling studies indicate that the ease of spontaneous transformation of mouse cells in culture may be due to a chaotic progression of aneuploidy. Conversely, the strong preference towards senescence and resistance to transformation of human cells in culture may be the result of a non-chaotic progression of aneuploidy. Finally, a method is proposed for quantifying the aneuploidogenic potencies of carcinogens. PMID- 10839978 TI - Retinoid-binding proteins: mediators of retinoid action. AB - Active vitamin A metabolites, known as retinoids, are essential for multiple physiological processes, ranging from vision to embryonic development. These small hydrophobic compounds associate in vivo with soluble proteins that are present in a variety of cells and in particular extracellular compartments, and which bind different types of retinoids with high selectivity and affinity. Traditionally, retinoid-binding proteins were viewed as transport proteins that act by solubilizing and protecting their labile ligands in aqueous spaces. It is becoming increasingly clear, however, that, in addition to this general role, retinoid-binding proteins have diverse and specific functions in regulating the disposition, metabolism and activities of retinoids. Some retinoid-binding proteins appear to act by sequestering their ligands, thereby generating concentration gradients that allow cells to take up retinoids from extracellular pools and metabolic steps to proceed in energetically unfavourable directions. Other retinoid-binding proteins regulate the metabolic fates of their ligands by protecting them from some enzymes while allowing metabolism by others. In these cases, delivery of a bound retinoid from the binding protein to the 'correct' enzyme is likely to be mediated by direct and specific interactions between the two proteins. One retinoid-binding protein was reported to enhance the ability of its ligand to regulate gene transcription by directly delivering this retinoid to the transcription factor that is activated by it. 'Channelling' of retinoids between their corresponding binding protein and a particular protein target thus seems to be a general theme through which some retinoid-binding proteins exert their effects. PMID- 10839980 TI - Glycosylation-site-selective synthesis of N-acetyl-lactosamine repeats in bis glycosylated human lysozyme. AB - We have studied the elongation of oligosaccharides containing N-acetyl lactosamine repeats using glycosylated human lysozyme mutants as a model. We reported previously that a combination of glycosylation sites at the 49th (site IV) and 68th (site II) amino acid residues of the protein particularly stimulates the synthesis of N-acetyl-lactosamine repeats [Melcher, Grosch, Grosse and Hasilik (1998) Glycoconjugate J. 15, 987-993]. In the present study we show that it is the carbohydrate attached to site IV that is selectively affected. It contains more N-acetyl-lactosamine repeats when site II is glycosylated in the same molecule. As a corollary of the glycosylation at site II, the synthesis of a third antenna at site IV is increased. The triantennary oligosaccharides at site IV contain more N-acetyl-lactosamine repeats than the biantennary ones. Thus placing a carbohydrate at site II stimulates the branching and the elongation of the carbohydrate at the other site. PMID- 10839981 TI - Multiple conserved segments of E1 small nucleolar RNA are involved in the formation of a ribonucleoprotein particle in frog oocytes. AB - E1/U17 small nucleolar RNA (snoRNA) is a box H/ACA snoRNA. To identify E1 RNA elements required for its assembly into a ribonucleoprotein (RNP) particle, we have made substitution mutations in evolutionarily conserved sequences and structures of frog E1 RNA. After E1 RNA was injected into the nucleus of frog oocytes, assembly of this exogenous RNA into an RNP was monitored by non denaturing gel electrophoresis. Unexpectedly, nucleotide substitutions in many phylogenetically conserved segments of E1 RNA produced RNPs with abnormal gel electrophoresis patterns. These RNA segments were at least nine conserved sequences and an apparently conserved structure. In another region needed for RNP formation, the requirement may be sequence(s) and/or structure. Base substitutions in each of these and in one additional conserved E1 RNA segment reduced the stability of this snoRNA in frog oocytes. Nucleolar localization was assayed by fluorescence microscopy after injection of fluorescein-labelled RNA. The H box (ANANNA) and the ACA box are both needed for efficient nucleolar localization of frog E1 RNA. PMID- 10839982 TI - Involvement of cytosolic phospholipase A2, and the subsequent release of arachidonic acid, in signalling by rac for the generation of intracellular reactive oxygen species in rat-2 fibroblasts. AB - Although there have been a number of recent studies on the role of Rac in the generation of reactive oxygen species (ROS), details of the signalling pathway remain unclear. In the present study we analysed the extent to which the activation of cytosolic phospholipase A(2) and the resultant release of arachidonic acid (AA) are involved in the Rac-mediated generation of ROS. Transfection of Rat-2 cells with RacV12, a constitutively active form of Rac1, induced elevated levels of ROS, as reflected by increased H(2)O(2)-sensitive fluorescence of 2', 7'-dichlorofluorescein. These effects could be blocked by inhibiting phospholipase A(2) or 5-lipoxygenase but not by inhibiting cyclo oxygenase. The application of exogenous AA increased levels of ROS but the effect was dependent on the further metabolism of AA to leukotrienes C(4)/D(4)/E(4) by 5 lipoxygenase. Indeed, the exogenous application of a mixture of leukotrienes C(4)/D(4)/E(4) elicited transient elevations in the levels of ROS that were blocked by catalase. These findings indicate that phospholipase A(2) and subsequent AA metabolism by 5-lipoxygenase act as downstream mediators in a Rac signalling pathway leading to the generation of ROS. PMID- 10839983 TI - G-protein coupling of mu-opioid receptors (OP3): elevated basal signalling activity. AB - To determine mu-opioid receptor (OP(3)) signalling activity, guanosine 5'-[gamma [(35)S]thio]triphosphate (GTP[(35)S]) binding to G-proteins was measured in the membranes of human embryonic kidney cells (HEK-293) transfected with mu-opioid receptor (HEK-mu). GTP[(35)S] binding to HEK-mu membranes was significantly elevated compared with HEK-293 control membranes (without OP(3)), and this was abolished by pertussis-toxin pretreatment. The irreversible antagonist beta chlornaltrexamine (beta-CNA) dose-dependently decreased elevated basal G-protein coupling of HEK-mu to control levels in cells devoid of OP(3). This characterizes beta-CNA as an inverse OP(3) agonist. Immunoprecipitation of solubilized G proteins with G(i3)alpha antisera demonstrated that basal GTP[(35)S] binding to G(i3)alpha was also substantially elevated in HEK-mu membranes over the control, whereas G(i3)alpha protein levels were unchanged. Basal GTP[(35)S] binding to G(i1)alpha/G(i2)alpha and G(o)alpha was also increased twofold in HEK-mu membranes over the control. Morphine further increased coupling to each of these Galpha proteins with similar potency, but not to G(q)/(11)alpha or G(s)alpha. These results indicate that the wild-type OP(3) can couple constitutively to endogenously expressed G(i3)alpha, G(i1)alpha/G(i2)alpha and G(o)alpha subunits of G-proteins in HEK-293 cells. PMID- 10839984 TI - A novel lysine 2,3-aminomutase encoded by the yodO gene of bacillus subtilis: characterization and the observation of organic radical intermediates. AB - The yodO gene product of Bacillus subtilis has been cloned and overexpressed in Escherichia coli and purified. The nucleotide sequence encodes a protein of 471 amino acids with a calculated molecular mass of 54071 Da. The translated amino acid sequence is more than 60% identical to that of the lysine 2,3-aminomutase from Clostridium subterminale SB4. Analytical HPLC gel-permeation chromatography leads to an estimate of an over all molecular mass of 224000+/-21000 Da, which corresponds to a tetrameric protein. The purified protein contains iron, sulphide and pyridoxal 5'-phosphate (PLP) and displays an optical absorption band extending to 700 nm, suggesting the presence of an iron-sulphide cluster. After reductive incubation with L-cysteine anaerobically, the protein catalyses the transformation of L-lysine into beta-lysine in the presence of S adenosylmethionine (AdoMet) and sodium dithionite. The K(m) value for L-lysine is estimated to be 8.0+/-2.2 mM. The iron-sulphur centre is stable in air,allowing aerobic purification. EPR spectroscopy at 10 K of the purified enzyme revealed an EPR signal similar to that of the [4Fe-4S](3+) cluster observed in the clostridial lysine 2, 3-aminomutase. Incubation with cysteine under anaerobic conditions converts the iron-sulphur centre into the EPR-silent [4Fe-4S](2+). Unlike the clostridial enzyme, the fully reduced [4Fe-4S](+) could not be characterized by further reduction with dithionite in the presence of AdoMet, although both dithionite and AdoMet were required to activate the enzyme. Upon addition of L-lysine, dithionite and AdoMet to the reduced enzyme and freezing the solution to 77 K, the EPR spectrum revealed the presence of an organic free radical signal (g=2.0023), which displayed multiple hyperfine transitions very similar to the spectrum of the beta-lysine-related radical in the mechanism of the clostridial lysine 2,3-aminomutase. Experiments with isotopically substituted L-lysine and lysine analogues verified the association of spin density with the carbon skeleton of lysine. The data indicate that the protein encoded by the yodO gene of B. subtilis is a novel lysine 2,3-aminomutase. The E. coli homologue of clostridial lysine 2,3-aminomutase was also expressed in E. coli and purified. This protein contained ironand sulphide but not PLP, it did not display lysine 2,3-aminomutase activity, and addition of PLP did not induce 2,3-aminomutase activity. PMID- 10839986 TI - Atomic force microscopy of gastric mucin and chitosan mucoadhesive systems. AB - Atomic force microscopy has been utilized to probe, at a molecular level, the interaction between purified pig gastric mucin (PGM) and a mucoadhesive cationic polymer, chitosan (sea cure 210+), with a low degree (approx. 11%) of acetylation. Images were produced detailing the structures of both PGM and chitosan in 0.1 M acetate buffer (pH 4.5), followed by the complex of the two structures in the same buffer. PGM in 0.1 M acetate buffer revealed long linear filamentous structures, consistent with earlier electron microscopy and scanning tunnelling micoscopy studies. The chitosan molecules also adopted a linear conformation in the same buffer, although with a smaller average length and diameter. They appeared to adopt a stiff-coil conformation consistent with earlier hydrodynamic measurements. The complexes formed after mixing PGM and chitosan together revealed large aggregates. In 0.1 M ionic strength buffer they were of the order of 0.7 microm in diameter, consistent with previous electron microscopy studies. The effect of ionic strength of the buffer on the structure of the complex was also studied and, together with molecular hydrodynamic data, demonstrates that the interaction is principally electrostatic in nature. PMID- 10839985 TI - Ligand binding directly stimulates ubiquitination of the inositol 1, 4,5 trisphosphate receptor. AB - Down-regulation of the Ins(1,4,5)P(3) receptor is an adaptive response to the activation of certain phosphoinositidase C-linked cell-surface receptors. It is manifested as a profound decline in cellular Ins(1,4,5)P(3) receptor content, occurs with a half-time of 0.5-2 h and is due to accelerated proteolysis. It has been shown that this process is mediated by the ubiquitin/proteasome pathway and is therefore initiated by Ins(1,4,5)P(3) receptor ubiquitination. To investigate the role of ligand binding in Ins(1,4,5)P(3) receptor ubiquitination, we expressed 'exogenous' wild-type and ligand-binding-defective mutant type I Ins(1,4,5)P(3) receptors in human neuroblastoma SH-SY5Y cells, in which muscarinic receptor activation elicits Ins(1,4,5)P(3) receptor down-regulation. We found (1) that exogenous wild-type Ins(1,4,5)P(3) receptors are efficiently ubiquitinated in response to muscarinic receptor stimulation, (2) that exogenous ligand binding-defective mutant Ins(1,4,5)P(3) receptors are resistant to ubiquitination, (3) that this resistance is not caused by the removal of potential ubiquitin-conjugating sites in the mutated region, and (4) that in heterotetramers of exogenous mutant receptors and 'endogenous' receptors, only the latter are targeted for ubiquitination. These results indicate that the binding of Ins(1,4,5)P(3) directly stimulates ubiquitination of the Ins(1,4,5)P(3) receptor and that the targeting of Ins(1,4,5)P(3) receptors for ubiquitination is a highly specific process. We therefore propose that an Ins(1,4, 5)P(3)-binding-induced conformational change in the receptor exposes a degradation signal that leads to ubiquitination. PMID- 10839987 TI - Annexin II is associated with mRNAs which may constitute a distinct subpopulation. AB - Protein-mRNA interactions affect mRNA transport, anchorage, stability and translatability in the cytoplasm. During the purification of three subpopulations of polysomes, it was observed that a 36-kDa protein, identified as annexin II, is associated with only one specific population of polysomes, namely cytoskeleton associated polysomes. This association appears to be calcium-dependent since it was sensitive to EGTA and could be reconstituted in vitro. UV irradiation resulted in partial, EGTA-resistant cross-linking of annexin II to the polysomes. Binding of (32)P-labelled total RNA to proteins isolated from the cytoskeleton bound polysomes on a NorthWestern blot resulted in a radioactive band having the same mobility as annexin II and, most importantly, purified native annexin II immobilized on nitrocellulose specifically binds mRNA. The mRNA population isolated from cytoskeleton-bound polysomes binds to annexin II with the highest affinity as compared with those isolated from free or membrane-bound polysomes. Interestingly, the annexin II complex, isolated from porcine small intestinal microvilli was a far better substrate for mRNA binding than the complex derived from transformed Krebs II ascites cells. When cytoskeleton-associated polysomes were split into 60 S and 40 S ribosomal subunits, and a peak containing mRNA complexes, annexin II fractionated with the mRNAs. Finally, using affinity purification of mRNA on poly(A)(+)-coupled magnetic beads, annexin II was only detected in association with messenger ribonucleoproteins (mRNPs) present in the cytoskeletal fraction (non-polysomal mRNPs). These results, derived from both in vitro experiments and cell fractionation, suggest that annexin II binds directly to the RNA moiety of mRNP complexes containing a specific population of mRNAs. PMID- 10839988 TI - Enzymic methylation of arginyl residues in -gly-arg-gly- peptides. AB - N(G)-Methylation of arginine residues in many nucleic-acid-binding proteins are formed post-translationally, catalysed by S-adenosylmethionine:protein-arginine N methyltransferase in their glycine-rich and arginine-rich motifs. The amino acid sequences of the stimulator of HIV-1 TAR (Tat-responsive element) RNA-binding protein (SRB) and fibronectin also show the presence of the internal -Gly-Arg-Gly (-GRG-) sequence, which is potentially methylatable by the methyltransferase. To investigate the sequence requirement for methylation of these proteins, several synthetic oligopeptides with different chain lengths and sequences similar to the -GRG- regions of SRB and fibronectin were synthesized. Whereas the heptapeptide AGGRGKG (residues 16-22 in SRB) served as the methyl acceptor for the methyltransferase with a K(m) of 50 microM, the 19-mer peptide (residues 10-28 in SRB) was methylated with a K(m) of 8.3 microM, indicating that a greater peptide chain length yields a better methyl acceptor. Product analysis of the methylated [methyl-(14)C]SRB-peptide by HPLC indicated the formation of N(G) monomethylarginine and N(G),N(G)-dimethyl(asymmetric)arginine. Synthetic peptides containing the cell attachment sequence [Arg-Gly-Asp ('RGD')] in fibronectin, GRGDSPK, GGRGDSPK and GGGRGDSPK, were also studied; whereas GRGDSPK was a poor methyl acceptor, the longer peptides were better methyl acceptors. To provide an understanding of the effect of methylation on fibronectin peptide, arginine unmethylated and methylated GGRGDSPK were compared for their effect on the mitogenesis induced by beta-hexosaminidase A and an agonistic antibody (mAb(15)) in bovine tracheal smooth-muscle cells; whereas the former inhibited 35-67% of mitogenesis at a concentration of 5-10 microM, the latter did not block mitogenesis. This lack of inhibition by the insertion of a methyl group on the arginyl residue of the cell attachment sequence might be due to the hindrance of the binding of fibronectin peptide to integrins. PMID- 10839989 TI - Contrasting effects of N5-substituted tetrahydrobiopterin derivatives on phenylalanine hydroxylase, dihydropteridine reductase and nitric oxide synthase. AB - Tetrahydrobiopterin [(6R)-5,6,7,8-tetrahydro-L-biopterin, H(4)biopterin] is one of several cofactors of nitric oxide synthases (EC 1.14.13.39). Here we compared the action of N(5)-substituted derivatives on recombinant rat neuronal nitric oxide synthase with their effects on dihydropteridine reductase (EC 1.6.99.7) and phenylalanine hydroxylase (EC 1.14.16.1),the well-studied classical H(4)biopterin dependent reactions. H(4)biopterin substituted at N(5) with methyl, hydroxymethyl, formyl and acetyl groups were used. Substitution at N(5) occurs at a position critical to the redox cycle of the cofactor in phenylalanine hydroxylase/dihydropteridine reductase. We also included N(2)'-methyl H(4)biopterin, a derivative substituted at a position not directly involved in redox cycling, as a control. As compared with N(5)-methyl H(4)biopterin, N(5) formyl H(4)biopterin bound with twice the capacity but stimulated nitric oxide synthase to a lesser extent. Depending on the substituent used, N(5)-substituted derivatives were redox-active: N(5)-methyl- and N(5)-hydroxyl methyl H(4)biopterin, but not N(5)-formyl- and N(5)-acetyl H(4)biopterin, reduced 2,6 dichlorophenol indophenol. N(5)-Substituted H(4)biopterin derivatives were not oxidized to products serving as substrates for dihydropteridine reductase and,depending on the substituent, were competitive inhibitors of phenylalanine hydroxylase: N(5)-methyl- and N(5)-hydroxymethyl H(4)biopterin inhibited phenylalanine hydroxylase, whereas N(5)-formyl- and N(5)-acetyl H(4)biopterin had no effect. Our data demonstrate differences in the mechanism of stimulation of phenylalanine hydroxylase and nitric oxide synthase by H(4)biopterin. They are compatible with a novel, non-classical, redox-active contribution of H(4)biopterin to the catalysis of the nitric oxide synthase reaction. PMID- 10839990 TI - A new method for the selection of protein interactions in mammalian cells. AB - In the present study we present a new method that allows for the selection of protein interactions in mammalian cells. We have used this system to verify two interactions previously characterized in vitro. (1) The interaction between human TATA-binding protein 1 and nuclear factor kappaB and (2) the association of Homo sapiens nuclear autoantigen SP100B with human heterochromatin protein 1alpha, a protein implicated in chromatin remodelling. We observe for the first time that these interactions also occur in vivo. One protein was fused to the N-terminal half of ubiquitin, while the interacting partner was fused to the C-terminal half of ubiquitin, that was itself linked to guanine phosphoryltransferase 2 (gpt2) modified to begin with an arginine residue. Upon interaction of both proteins, ubiquitin is reconstituted, and its association with the Rgpt2 reporter is subsequently cleaved off by ubiquitin-processing enzymes. The presence of arginine in the Rgpt2 gene product leads to the degradation of the product by the N-end rule pathway. In the human fibroblast cell line HT1080HPRT(-) (that is deficient in the enzyme for hypoxanthine-guanine phosphoribosyltransferase) cells in which interaction between both proteins of interest occurs can then be selected for by hypoxanthine/aminopterin/thymine medium and counterselected against by 6-thioguanine medium. This method provides a suitable alternative to the yeast two-hybrid system and is generally applicable. PMID- 10839991 TI - Repression of transforming-growth-factor-beta-mediated transcription by nuclear factor kappaB. AB - Activation of transforming growth factor-beta (TGF-beta) and activin receptors leads to phosphorylation of Sma- and Mad-related protein 2 (Smad2) and Smad3, which function as transcription factors to regulate gene expression. Smad7 is a regulatory protein which is able to inhibit TGF-beta and activin signalling in a negative-feedback loop, mediated by a direct regulation by Smad3 and Smad4 via a Smad-binding element (SBE) in the Smad7 promoter. Interestingly, we found that the Smad7 promoter was also regulated by nuclear factor kappaB (NF-kappaB), a transcription factor which plays an important role in inflammation and the immune response. Expression of NF-kappaB p65 subunit was able to inhibit the Smad7 promoter activity, and this inhibition could be reversed by co-expression of IkappaB, an inhibitor of NF-kappaB. In addition, the inhibitory activity of p65 was observed in a minimal promoter that contained only the Smad7 SBE and a TATA box, without any consensus NF-kappaB binding site. This inhibitory effect appeared to be common to other TGF-beta- and activin-responsive promoters, since p65 also inhibited the forkhead-activin-signal-transducer-2-mediated activation of a Xenopus Mix.2 promoter, as well as the Smad3-mediated activation of 3TP-lux which contains PMA-responsive elements and a plasminogen-activator-inhibitor-1 promoter. Activation of endogenous NF-kappaB by tumour necrosis factor-alpha (TNF alpha) was also able to inhibit the Smad7 promoter in human embryonic kidney 293 cells. In human hepatoma HepG2 cells, TNF-alpha was able to inhibit TGF-beta- and activin-mediated transcriptional activation. Furthermore, overexpression of the transcription co-activator p300 could abrogate the inhibitory effect of NF-kappaB on the Smad7 promoter. Taken together, these data have indicated a novel mode of crosstalk between the Smad and the NF-kappaB signalling cascades at the transcriptional level by competing for a limiting pool of transcription co activators. PMID- 10839992 TI - Determinant of the extracellular location of the N-terminus of human multidrug resistance-associated protein. AB - Multidrug-resistance-associated protein (MRP) is a member of the ATP-binding cassette (ABC) membrane-transport superfamily and is responsible for multidrug resistance in cancer cells. Distinct from other members of the ABC superfamily, MRP has three membrane-spanning domains (MSDs) and the N-terminus is located extracellularly. It has been shown that the first MSD (MSD1) with an extracellular N-terminus is important for MRP function. To address what ensures the generation of this structural organization of MRP and to understand in general the molecular mechanism of membrane folding of polytopic proteins with extracellular N-termini, the biogenesis of MSD1 in human MRP1 was examined using an in vitro expression system. Surprisingly, the second transmembrane segment (TM2) in MSD1 was found to play a critical role in the correct membrane translocation and folding of MSD1 in human MRP1. TM2 not only plays an essential role to ensure the N-terminus-outside/C-terminus-inside orientation of TM1 with an extracellular N-terminus, it can also translocate into membranes post translationally in a signal-recognition particle and ribosome-dependent manner to provide an additional insurance for correct folding of MSD1 in MRP. These findings suggest that TM2 in a polytopic membrane protein with an extracellular N terminus may play a critical role in controlling correct membrane translocation and folding of the protein in general. PMID- 10839993 TI - Evidence that metformin exerts its anti-diabetic effects through inhibition of complex 1 of the mitochondrial respiratory chain. AB - Although metformin is widely used for the treatment of non-insulin-dependent diabetes, its mode of action remains unclear. Here we provide evidence that its primary site of action is through a direct inhibition of complex 1 of the respiratory chain. Metformin(50 microM) inhibited mitochondrial oxidation of glutamate+malate in hepatoma cells by 13 and 30% after 24 and 60 h exposure respectively, but succinate oxidation was unaffected. Metformin also caused time dependent inhibition of complex 1 in isolated mitochondria, whereas in sub mitochondrial particles inhibition was immediate but required very high metformin concentrations (K(0.5),79 mM). These data are compatible with the slow membrane potential-driven accumulation of the positively charged drug within the mitochondrial matrix leading to inhibition of complex 1. Metformin inhibition of gluconeogenesis from L-lactate in isolated rat hepatocytes was also time- and concentration-dependent, and accompanied by changes in metabolite levels similar to those induced by other inhibitors of gluconeogenesis acting on complex 1. Freeze-clamped livers from metformin-treated rats exhibited similar changes in metabolite concentrations. We conclude that the drug's pharmacological effects are mediated, at least in part, through a time-dependent, self-limiting inhibition of the respiratory chain that restrains hepatic gluconeogenesis while increasing glucose utilization in peripheral tissues. Lactic acidosis, an occasional side effect, canal so be explained in this way. PMID- 10839994 TI - Dynamics of haem oxygenase-1 expression and bilirubin production in cellular protection against oxidative stress. AB - The inducible isoform of haem oxygenase (HO-1) has been proposed as an effective system to counteract oxidant-induced cell injury. In several circumstances, this cytoprotective effect has been attributed to increased generation of the antioxidant bilirubin during haem degradation by HO-1. However, a direct implication for HO-1-derived bilirubin in protection against oxidative stress remains to be established. In the present study, we examined the dynamics of HO-1 expression and bilirubin production after stimulation of vascular smooth-muscle cells with hemin, a potent inducer of the HO-1 gene. We found that hemin-mediated increase in HO-1 protein expression and haem oxygenase activity is associated with augmented bilirubin levels. The majority of bilirubin production occurred early after exposure of cells to hemin. Hemin pre-treatment also resulted in high resistance to cell injury caused by an oxidant-generating system. Interestingly, this protective effect was manifest only when cells were actively producing bilirubin as a consequence of increased haem availability and utilization by HO 1. Tin protoporphyrin IX, an inhibitor of haem oxygenase activity, significantly reduced bilirubin generation and reversed cellular protection afforded by hemin treatment. Furthermore, addition of bilirubin to the culture medium markedly reduced the cytotoxicity produced by oxidants. Our findings provide direct evidence that bilirubin generated after up-regulation of the HO-1 pathway is cytoprotective against oxidative stress. PMID- 10839995 TI - Characterization of beta-galactosidase mutations Asp332-->Asn and Arg148-->Ser, and a polymorphism, Ser532-->Gly, in a case of GM1 gangliosidosis. AB - We have identified and characterized three missense mutations in a patient with type 1 G(M1) gangliosidosis, namely a substitution of G for A at nucleotide position 1044 (G1044-->A; in exon 10) on one allele, which converts Asp(332) into asparagine, and both a mutation (C492-->A in exon 4, leading to the amino acid change of Arg(148)-->Ser) and a polymorphism (A1644-->G in exon 15, leading to a change of Ser(532)-->Gly) on the other allele. This patient had less than 1% residual beta-galactosidase activity and minimally detectable levels of immunoreactive beta-galactosidase protein in fibroblasts. To account for the above findings, a series of expression and immunolocalization studies were undertaken to assess the impact of each mutation. Transient overexpression in COS 1 cells of cDNAs encoding Asp(332)Asn, Arg(148)Ser and Ser(532)Gly mutant beta galactosidases produced abundant amounts of precursor beta-galactosidase, with activities of 0, 84 and 81% compared with the cDNA clone for wild-type beta galactosidase (GP8). Since the level of vector-driven expression is much less in Chinese hamster ovary (CHO) cells than in COS-1 cells, and we knew that exogenous beta-galactosidase undergoes lysosomal processing when expressed in these cells, transient expression studies were performed of Arg(148)Ser and Ser(532)Gly, which yielded active forms of the enzyme. In this case, the Arg(148)Ser and Ser(532)Gly products gave rise to 11% and 86% of the control activity respectively. These results were not unexpected, since the Arg(148)Ser mutation introduced a major conformational change into the protein, and we anticipated that it would be degraded in the endoplasmic reticulum (ER), whereas the polymorphism was expected to produce near-normal activity. To examine the effect of the Asp(332)Asn mutation on the catalytic activity, we isolated CHO clones permanently transfected with the Asp(332)Asn and Asp(332)Glu constructs, purified the enzymes by substrate-analogue-affinity chromatography, and determined their kinetic parameters. The V(max) values of both mutant recombinant enzymes were markedly reduced (less than 0.9% of the control), and the K(m) values were unchanged compared with the corresponding wild-type enzyme isolated at the same time. Both the Arg(148)Ser beta-galactosidase in CHO cells and Asp(332)Asn beta galactosidases (in COS-1 and CHO cells) produced abundant immunoreaction in the perinuclear area, consistent with localization in the ER. A low amount was detected in lysosomes. Incubation of patient fibroblasts in the presence of leupeptin, which reduces the rate of degradation of lysosomal beta-galactosidase by thiol proteases, had no effect on residual enzyme activity, and immunostaining was again detected largely in the perinuclear area (localized to the ER) with much lower amounts in the lysosomes. In summary, the Arg(148)Ser mutation has no effect on catalytic activity, whereas the Asp(332)Asn mutation seriously reduces catalytic activity, suggesting that Asp(332) might play a role in the active site. Immunofluorescence studies indicate the expressed mutant proteins with Arg(148)Ser and Asp(332)Asn mutations are held up in the ER, where they are probably degraded, resulting in only minimum amounts of the enzyme becoming localized in the lysosomes. These results are completely consistent with findings in the cultured fibroblasts. Our results imply that most of the missense mutations described in G(M1) gangliosidosis to date have little effect on catalytic activity, but do affect protein conformation such that the resulting protein cannot be transported out of the ER and fails to arrive in the lysosome. This accounts for the minimal amounts of enzyme protein and activity seen in most G(M1) gangliosidosis patient fibroblasts. PMID- 10839996 TI - Possible interference between tissue-non-specific alkaline phosphatase with an Arg54-->Cys substitution and acounterpart with an Asp277-->Ala substitution found in a compound heterozygote associated with severe hypophosphatasia. AB - Tissue-non-specific alkaline phosphatase (TNSALP) with an Arg(54)-->Cys (R54C) or an Asp(277)-->Ala (D277A)substitution was found in a patient with hypophosphatasia [Henthorn,Raducha, Fedde, Lafferty and Whyte (1992) Proc. Natl. Acad. Sci. U.S.A.89, 9924-9928]. To examine effects of these missense mutations onproperties of TNSALP, the TNSALP mutants were expressed ectopically inCOS-1 cells. The wild-type TNSALP was synthesized as a 66-kDa endo-beta-N acetylglucosaminidase H (Endo H)-sensitive form, and processed to an 80-kDa mature form, which is anchored to the plasma membrane via glycosylphosphatidylinositol (GPI). Although the mutant proteins were found to be modified by GPI, digestion with phosphatidylinositol-specific phospholipase C, cell-surface biotinylation and immunofluorescence observation demonstrated that the cell-surface appearance of TNSALP (R54C) and TNSALP (D277A) was either almost totally or partially retarded respectively. The 66-kDa Endo H-sensitive band was the only form, and was rapidly degraded in the cells expressing TNSALP (R54C). In contrast with cells expressing TNSALP(R54C), where alkaline phosphatase activity was negligible, significant enzyme activity was detected and, furthermore, the 80 kDa mature form appeared on the surface of the cells expressing TNSALP (D277A). Analysis by sedimentation on sucrose gradients showed that a considerable fraction of newly synthesized TNSALP (R54C) and TNSALP(D277A) formed large aggregates, indicating improper folding and incorrect oligomerization of the mutant enzymes. When co-expressed with TNSALP (R54C), the level of the 80-kDa mature form of TNSALP (D277A)was decreased dramatically, with a concomitant reduction in enzyme activity in the co-transfected cell. These findings suggest that TNSALP(R54C) interferes with folding and assembly of TNSALP (D277A) intrans when expressed in the same cell, thus probably explaining why a compound heterozygote for these mutant alleles developed severe hypophosphatasia. PMID- 10839997 TI - The molecular basis of phosphatidylcholine preference of human group-V phospholipase A2. AB - Human group-V phospholipase A(2) (hVPLA(2)) is a secretory phospholipase A(2) (PLA(2)) that is involved in eicosanoid formation in such inflammatory cells as macrophages and mast cells. We showed that hVPLA(2) can bind phosphatidylcholine membranes and hydrolyse phosphatidylcholine molecules much more efficiently than human group-IIa PLA(2), which accounts for its high activity on the outer plasma membrane of mammalian cells. To understand the molecular basis of the high phosphatidylcholine specificity of hVPLA(2), we mutated several residues (Gly-53, Glu-56 and Glu-57) that might be involved in interaction with an active-site bound phospholipid molecule. Phospholipid head-group specificities of mutants determined using polymerized mixed-liposome substrates indicate that a small glycine residue in position 53 is important for accommodating a bulky choline head group. Also, results indicated that two anionic residues, Glu-56 and Glu-57, favourably interact with cationic head groups of phosphatidylcholine and phosphatidylethanolamine. Together, these steric and electrostatic properties of the active site of hVPLA(2) allow for effective binding and hydrolysis of a bulky cationic choline head group of phosphatidylcholine, which is unique among mammalian secretory PLA(2)s. PMID- 10839998 TI - Defensin-like peptide-2 from platypus venom: member of a class of peptides with a distinct structural fold. AB - The venom of the male Australian duck-billed platypus contains a family of four polypeptides of appox. 5 kDa, which are referred to as defensin-like peptides (DLPs). They are unique in that their amino acid sequences have no significant similarities to those of any known peptides; however, the tertiary structure of one of them, DLP-1, has recently been shown to be similar to beta-defensin-12 and to the sodium neurotoxin peptide ShI (Stichodactyla helianthus neurotoxin I). Although DLPs are the major peptides in the platypus venom, little is known about their biological roles. In this study, we determined the three-dimensional structure of DLP-2 by NMR spectroscopy, with the aim of gaining insights into the natural function of the DLPs in platypus venom. The DLP-2 structure was found to incorporate a short helix that spans residues 9-12, and an antiparallel beta sheet defined by residues 15-18 and 37-40. The overall fold and cysteine-pairing pattern of DLP-2 were found to be similar to those of DLP-1, and hence beta defensin-12; however, the sequence similarities between the three molecules are relatively small. The distinct structural fold of the DLP-1, DLP-2, and beta defensin-12 is based upon several key residues that include six cysteines. DLP-3 and DLP-4 are also likely to be folded similarly since they have high sequence similarity with DLP-2. The DLPs, and beta-defensin-12 may thus be grouped together into a class of polypeptide molecules which have a common or very similar global fold. The fact that the DLPs did not display antimicrobial, myotoxic, or cell-growth-promoting activities implies that the nature of the side chains in this group of peptides is likely to play an important role in defining the biological function(s). PMID- 10839999 TI - Modulation of the smooth-muscle L-type Ca2+ channel alpha1 subunit (alpha1C-b) by the beta2a subunit: a peptide which inhibits binding of beta to the I-II linker of alpha1 induces functional uncoupling. AB - Modulation of the smooth-muscle Ca(2+) channel alpha1C-b subunit by the auxiliary beta2a subunit was studied in the HEK 293 (cell line from human embryonic kidney cells) expression system. In addition, we tested whether the alpha1-beta interaction in functional channels is sensitive to an 18-amino-acid synthetic peptide that corresponds to the sequence of the defined major interaction domain in the cytoplasmic I-II linker of alpha1C (AID-peptide). Ca(2+) channels derived by co-expression of alpha1C-b and beta2a subunits exhibited an about 3-fold higher open probability (P(o)) than alpha1C-b channels. High-P(o) gating of alpha1C-b.beta2a channels was associated with the occurrence of long-lasting channel openings [mean open time (tau)>10 ms] which were rarely observed in alpha1C-b channels. Modulation of fast gating by the beta2a subunit persisted in the cell-free, inside-out recording configuration. Biochemical experiments showed that the AID-peptide binds with appreciable affinity to beta2 subunits of native Ca(2+) channels. Binding of the beta2 protein to immobilized AID-peptide was specifically inhibited (K(i) of 100 nM) by preincubation with free (uncoupled) AID-peptide, but not by a corresponding scrambled peptide. Administration of the AID-peptide (10 microM) to the cytoplasmic side of inside-out patches induced a substantial reduction of P(o) of alpha1C-b.beta2a channels. The scrambled control peptide failed to affect alpha1C-b. beta2a channels, and the AID-peptide (10 microM) did not modify alpha1C-b channel function in the absence of expressed beta2a subunit. Our results demonstrate that the beta2a subunit controls fast gating of alpha1C-b channels, and suggest the alpha1-beta interaction domain in the cytoplasmic I-II linker of alpha1C (AID) as a possible target of modulation of the channel. Moreover, our data are consistent with a model of alpha1-beta interaction that is based on multiple interaction sites, including AID as a determinant of the affinity of the alpha1-beta interaction. PMID- 10840000 TI - Activity of phosphatidylinositol transfer protein is sensitive to ethanol and membrane curvature. AB - Phosphatidylinositol transfer protein (PITP) is critical for many cellular signalling and trafficking events that are influenced by ethanol. The influence of ethanol and membrane curvature on the activity of recombinant mouse PITP-alpha in vitro is evaluated by monitoring the transfer of phosphatidylinositol (PtdIns) from rat hepatic microsomes to unilamellar vesicles. Acute exposure to pharmacological levels of ethanol enhanced the function of PITP. Chloroform shared a similar ability to enhance function when both drug concentrations were normalized to their respective octanol/water partition coefficients, indicating that the effect is not unique to ethanol and might be common to hydrophobic solutes. Neither the PITP activity nor its ethanol enhancement was altered by using thermally pretreated (denatured) or protease-treated microsomes, indicating that the native microsomal protein structure was unlikely to be a determinant of transfer. Kinetic analyses indicated that ethanol acted by increasing the PITP mediated flux of PtdIns from both microsomal and liposomal surfaces. The activity of PITP was strongly dependent on the lipid structure, with a steep dependence on the expressed curvature of the membrane. Activity was greatest for small, highly curved sonicated vesicles and decreased markedly for large, locally planar unilamellar vesicles. Ethanol enhanced PITP-mediated PtdIns transfer to all vesicles, but its effect was much smaller than the enhancement due to curvature, which is consistent with ethanol's comparatively modest ability to perturb membrane lipids. The ethanol efficacy observed is as pronounced as any previously described lipid-mediated ethanol action. In addition, these observations raise the possibility that PITP specifically delivers PtdIns to metabolically active membrane domains of convex curvature and/or low surface densities of lipid. PMID- 10840002 TI - June 6, 2000 PMID- 10840003 TI - Harold T. Dodge, MD PMID- 10840001 TI - Sequence of the 5'-flanking region and promoter activity of the human mucin gene MUC5B in different phenotypes of colon cancer cells. AB - Control of gene expression in intestinal cells is poorly understood. Molecular mechanisms that regulate transcription of cellular genes are the foundation for understanding developmental and differentiation events. Mucin gene expression has been shown to be altered in many intestinal diseases and especially cancers of the gastrointestinal tract. Towards understanding the transcriptional regulation of a member of the 11p15.5 human mucin gene cluster, we have characterized 3.55 kb of the 5'-flanking region of the human mucin gene MUC5B, including the promoter, the first two exons and the first intron. We report here the promoter activity of successively 5'-truncated sections of 956 bases of this region by fusing it to the coding region of a luciferase reporter gene. The transcription start site was determined by primer-extension analysis. The region upstream of the transcription start site is characterized by the presence of a TATA box at bases -32/-26, DNA-binding elements for transcription factors c-Myc, N-Myc, Sp1 and nuclear factor kappaB as well as putative activator protein (AP)-1-, cAMP response-element-binding protein (CREB)-, hepatocyte nuclear factor (HNF)-1-, HNF 3-, TGT3-, gut-enriched Kruppel factor (GKLF)-, thyroid transcription factor (TTF)-1- and glucocorticoid receptor element (GRE)-binding sites. Intron 1 of MUC5B was also characterized, it is 2511 nucleotides long and contains a DNA segment of 259 bp in which are clustered eight tandemly repeated GA boxes and a CACCC box that bind Sp1. AP-2alpha and GATA-1 nuclear factors were also shown to bind to their respective cognate elements in intron 1. In transfection studies the MUC5B promoter showed a cell-specific activity as it is very active in mucus secreting LS174T cells, whereas it is inactive in Caco-2 enterocytes and HT-29 STD (standard) undifferentiated cells. Within the promoter, maximal transcription activity was found in a segment covering the first 223 bp upstream of the transcription start site. Finally, in co-transfection experiments a transactivating effect of Sp1 on to MUC5B promoter was seen in LS174T and Caco-2 cells. PMID- 10840004 TI - Echocardiographic destruction of albumin microbubbles directs gene delivery to the myocardium. AB - BACKGROUND: The noninvasive, tissue-specific delivery of therapeutic agents to the heart would be a valuable clinical tool. This study addressed the hypothesis that albumin-coated microbubbles could be used to effectively deliver an adenoviral transgene to rat myocardium by ultrasound-mediated microbubble destruction. METHODS AND RESULTS: Recombinant adenovirus containing beta galactosidase and driven by a constitutive promoter was attached to the surface of albumin-coated, perfluoropropane-filled microbubbles. These bubbles were infused into the jugular vein of rats with or without simultaneous echocardiography. Additional controls included ultrasound of microbubbles that did not contain virus, virus alone, and virus plus ultrasound. One group underwent ultrasound-mediated destruction of microbubbles followed by adenovirus infusion. Rats were killed after 4 days and examined for beta-galactosidase expression. The hearts of all rats that underwent ultrasound-mediated destruction of microbubbles containing virus showed nuclear staining with 5-bromo-4-chloro-3 indolyl-beta-D-galactopyranoside substrate, indicating expression of the transgene. None of the control animals showed myocardial expression of the beta galactosidase transgene. By quantitative analysis, beta-galactosidase activity was 10-fold higher in the treated group than in controls (P<0.0001). CONCLUSIONS: Ultrasound-mediated destruction of albumin-coated microbubbles is a promising method for the delivery of bioactive agents to the heart. PMID- 10840005 TI - Predictors of outcome in patients with acute coronary syndromes without persistent ST-segment elevation. Results from an international trial of 9461 patients. The PURSUIT Investigators. AB - BACKGROUND: Appropriate treatment policies should include an accurate estimate of a patient's baseline risk. Risk modeling to date has been underutilized in patients with acute coronary syndromes without persistent ST-segment elevation. METHODS AND RESULTS: We analyzed the relation between baseline characteristics and the 30-day incidence of death and the composite of death or myocardial (re)infarction in 9461 patients with acute coronary syndromes without persistent ST-segment elevation enrolled in the PURSUIT trial [Platelet glycoprotein IIb/IIIa in Unstable angina: Receptor Suppression Using Integrilin (eptifibatide) Therapy]. Variables examined included demographics, history, hemodynamic condition, and symptom duration. Risk models were created with multivariable logistic regression and validated by bootstrapping techniques. There was a 3.6% mortality rate and 11.4% infarction rate by 30 days. More than 20 significant predictors for mortality and for the composite end point were identified. The most important baseline determinants of death were age (adjusted chi(2)=95), heart rate (chi(2)=32), systolic blood pressure (chi(2)=20), ST-segment depression (chi(2)=20), signs of heart failure (chi(2)=18), and cardiac enzymes (chi(2)=15). Determinants of mortality were generally also predictive of death or myocardial (re)infarction. Differences were observed, however, in the relative prognostic importance of predictive variables for mortality alone or the composite end point; for example, sex was a more important determinant of the composite end point (chi(2)=21) than of death alone (chi(2)=10). The accuracy of the prediction of the composite end point was less than that of mortality (C index 0.67 versus 0.81). CONCLUSIONS: The occurrence of adverse events after presentation with acute coronary syndromes is affected by multiple factors. These factors should be considered in the clinical decision-making process. PMID- 10840006 TI - Relationship of Chlamydia pneumoniae infection to severity of human coronary atherosclerosis. AB - BACKGROUND: Infection with Chlamydia pneumoniae has been postulated to play a pathogenic role in atherosclerosis. We examined the role of infection with C pneumoniae in relation to the extent of coronary atherosclerosis. METHODS AND RESULTS: Coronary atherosclerosis was graded microscopically on a postmortem basis in a blinded fashion in 60 subjects as mild (n=18) or severe (n=42) atherosclerosis. Serum antibodies to C pneumoniae were measured by microimmunofluorescence test. Paraffin-embedded coronary artery specimens were examined for the presence of chlamydia by use of a genus-specific direct immunofluorescence monoclonal antibody. Frozen coronary artery specimens were examined by immunoperoxidase for the presence of C pneumoniae by use of a specific monoclonal antibody RR-402. Direct immunofluorescence was reactive in 86% of cases with severe atherosclerosis but in only 6% of cases with mild atherosclerosis (P<0.01), whereas immunoperoxidase staining was reactive in 80% and 38% of cases with severe and mild atherosclerosis, respectively (P<0. 01). Elevated IgG and IgA levels against C pneumoniae were not different in cases with severe and mild atherosclerosis (61% and 30% for severe atherosclerosis and 67% and 42% for mild atherosclerosis, respectively). CONCLUSIONS: This study supports the hypothesis that intracellular infection with C pneumoniae may relate to the severity of atherosclerosis in some subjects. Serum antibody titers against C pneumoniae do not differentiate between severe and mild atherosclerosis. PMID- 10840007 TI - Hormone replacement therapy and incidence of acute myocardial infarction. A population-based nested case-control study. AB - BACKGROUND: Epidemiological studies suggest a decreased risk of coronary heart disease (CHD) in healthy women taking hormonal replacement therapy (HRT). Whether this effect is shared by oral and transdermal preparations is unknown. METHODS AND RESULTS: We conducted a population-based case-control study nested in a cohort of women 50 to 74 years of age without cardiovascular disease history in the United Kingdom. Among 164 769 women from the General Practice Research Database (January 1, 1991, to December 31, 1995), we identified 1242 first acute myocardial infarctions (AMI) and confirmed 1013 after medical record review. We randomly selected 5000 age-frequency-matched control subjects. AMI incidence was 1.6 per 1000 person-years; 13% and 17% of cases and control subjects used HRT within 6 months before the index date. Risk factor and comorbidity-adjusted OR of AMI for current-recent HRT users compared with nonusers was 0.72 (95% CI 0.59 to 0.89). The OR was similar within 30 days before the index date. The beneficial effect was present after 1 year of use (OR 0. 68; 95% CI 0.53 to 0.86), with no increase in risk within the first year. ORs for unopposed and opposed therapy were 0.52 (95% CI 0.35 to 0.78) and 0.79 (95% CI 0. 59 to 1.08); 79% and 21% used oral and transdermal therapy. The protective effect was present at medium-high doses of estrogens with ORs for oral and transdermal therapy of 0.63 (95% CI 0.46 to 0.86) and 0.62 (95% CI 0.37 to 1.06) and ceased after 2 to 3 years since stopping HRT. CONCLUSIONS: Results are consistent with those previously reported in women without CHD who were taking oral HRT and, although based on few users, suggest that transdermal therapy might have similar cardioprotective effects. PMID- 10840008 TI - Extent of cardiac sympathetic neuronal damage is determined by the area of ischemia in patients with acute coronary syndromes. AB - BACKGROUND: Prior studies have demonstrated that acute ischemic injury causes sympathetic neuronal damage exceeding the area of necrosis. The aim of this study was to test the hypothesis that sympathetic neuronal damage measured by (123)I metaiodobenzylguanidine (MIBG) imaging would be determined by the area of ischemia as reflected by area at risk in patients undergoing reperfusion therapy for acute coronary syndromes. METHODS AND RESULTS: In 12 patients, the myocardium at risk was assessed by (99m)Tc-sestamibi SPECT before reperfusion, and infarct size was measured by follow-up (99m)Tc-sestamibi SPECT 1 week later. All patients also underwent (123)I-MIBG SPECT within a mean of 11 days after onset. The SPECT image analysis was based on a semiquantitative polar map approach. Defect size on the (123)I-MIBG or (99m)Tc-sestamibi SPECT was measured for the left ventricle (LV) with the use of a threshold of -2.5 SD from the mean value of a normal database and was expressed as %LV. The (123)I-MIBG defect size (47+/-18%LV) was larger than the infarct size (27+/-23%LV, P<0. 001) but was similar to the risk area (49+/-18%LV, P=NS). Furthermore, the (123)I-MIBG defect size was closely correlated with the risk area (r=0.905, P<0.001). CONCLUSIONS: Sympathetic neuronal damage measured by (123)I-MIBG SPECT is larger than infarct size and is closely related to risk area, suggesting high sensitivity of neuronal structures to ischemia compared with myocardial cells. PMID- 10840009 TI - Distinct patterns of dystrophin organization in myocyte sarcolemma and transverse tubules of normal and diseased human myocardium. AB - BACKGROUND: Genetic mutations of dystrophin and associated glycoproteins underlie cell degeneration in several inherited cardiomyopathies, although the precise physiological role of these proteins remains under discussion. We studied the distribution of dystrophin in relation to the force-transducing vinculin-rich costameres in left ventricular cardiomyocytes from normal and failing human hearts to further elucidate the function of this protein complex. METHODS AND RESULTS: Single- and double-label immunoconfocal microscopy and parallel high resolution immunogold fracture-label electron microscopy were used to localize dystrophin and vinculin in human left ventricular myocytes from normal (n=6) and failing hearts (idiopathic dilated cardiomyopathy, n=7, or ischemic heart disease, n=5). In control cardiomyocytes, dystrophin had a continuous distribution at the peripheral sarcolemma, with concentrated bands corresponding to the vinculin-rich costameres. Intracellular labeling extended along transverse (T) tubule membranes. Fracture-label confirmed this distribution, showing significantly greater label on plasma membrane fractures overlying I-bands (I band 4.1+/-0.3 gold particles/micrometer A-band 3.3+/-0.2 gold particles/micrometer mean+/-SE, P=0.02). Hypertrophied myocytes from failing hearts showed maintenance of this surface distribution except in degenerating cells; there was a clear increase in intracellular dystrophin label reflecting T tubule hypertrophy. CONCLUSIONS: Dystrophin partially colocalizes with costameric vinculin in normal and hypertrophied myocytes, a distribution lost in degenerating cells. This suggests a primarily mechanical role for dystrophin in maintenance of cell membrane integrity in normal and hypertrophied myocytes. The presence of dystrophin in the cardiac T-tubule membrane, in contrast to its known absence in skeletal muscle T-tubules, implies additional roles for dystrophin in membrane domain organization. PMID- 10840010 TI - Left ventricular concentric remodeling rather than left ventricular hypertrophy is related to the insulin resistance syndrome in elderly men. AB - BACKGROUND: Associations between left ventricular (LV) geometry and the insulin resistance syndrome have been found, mostly in small studies of middle-aged hypertensives. The purpose of this study was to elucidate these associations through the use of a large sample of elderly men. METHODS AND RESULTS: We investigated 475 men (157 hypertensives) 71 years of age who were attending a population-based health survey in Uppsala County with echocardiography, oral glucose tolerance test (OGTT), hyperinsulinemic euglycemic clamp, and lipid and 24-hour ambulatory blood pressure monitoring. LV relative wall thickness was significantly related to clamp insulin sensitivity index (r=-0.14), fasting insulin, 32-33 split proinsulin, triglycerides, nonesterified fatty acids, OGTT glucose and insulin levels, waist-to-hip ratio, body mass index, 24-hour blood pressure, and heart rate (r=0.10 to 0.22). Only 24-hour systolic pressure (r=0. 15), OGTT 2-hour insulin (r=-0.10), and heart rate (r=-0.14) were significantly related to LV mass index. Comparing subjects with various LV geometry (normal, concentric remodeling and concentric and eccentric hypertrophy) showed that 24 hour heart rate, OGTT glucose and insulin levels, waist-to-hip ratio, and body mass index were significantly higher (P<0.001 to 0.05) and clamp insulin sensitivity index was significantly lower (P<0.01) in the concentric remodeling geometry group than in the normal LV geometry group. The 24-hour blood pressure was significantly higher in the concentric hypertrophy group than in the normal LV geometry group (P<0.001). CONCLUSIONS: Several components of the insulin resistance syndrome were related to thick LV walls and concentric remodeling but less to LV hypertrophy in this population-based sample of elderly men. PMID- 10840011 TI - Local pulse pressure and regression of arterial wall hypertrophy during long-term antihypertensive treatment. AB - BACKGROUND: Local pulse pressure (PP) is an independent determinant of carotid artery wall thickness, stronger than mean blood pressure (BP). The present study was designed to assess whether a beta-adrenoceptor antagonist-based or an ACE inhibitor-based treatment was able to reduce carotid artery wall hypertrophy through a reduction in carotid PP rather than by lowering mean BP and whether the influence of local PP reduction could also be detected at the site of a muscular artery, the radial artery. METHODS AND RESULTS: Ninety-eight essential hypertensive patients were randomized to 9 months of double-blind treatment with either celiprolol or enalapril. Arterial parameters were determined with high resolution echo-tracking systems. PP was measured locally with applanation tonometry and independently of mean BP. After 9 months of treatment, mean BP, carotid PP, and intimal-medial thickness (IMT) decreased significantly, with no difference between the 2 groups. The reduction in carotid PP but not in mean BP was a major independent determinant of the reduction in carotid IMT. Radial artery IMT and PP decreased significantly with both treatments. However, the reduction in radial artery IMT was not related to the changes in radial artery PP. CONCLUSIONS: The regression of carotid artery wall hypertrophy during long term antihypertensive treatment was dependent on the reduction in local PP rather than on the lowering of mean BP. The effect of PP lowering on IMT reduction was observed at the site of an elastic artery but not at the site of a muscular artery. PMID- 10840012 TI - Effects of the maze operation on health-related quality of life in patients with atrial fibrillation. AB - BACKGROUND: Maze surgery for atrial fibrillation (AF) is a curative therapy, but its effect on health-related quality of life has not been studied. METHODS AND RESULTS: Maze operations were performed in 48 patients with drug-refractory AF. The majority of patients (80%) had lone AF, and the primary indication for surgery in all patients was AF. The SF-36 Health Survey was used to assess quality of life before operation and at 6 months and 1 year after surgery. Twenty five patients were available for the 1-year follow-up and completed all questionnaires. Before maze surgery, the SF-36 scores were significantly lower than in the general Swedish population, reflecting significant impairment in well being, physical and social functioning, and mental health. After maze surgery, the quality of life was significantly improved at 6 months and at 1 year on all scales except for bodily pain, which, however, was not significantly decreased before surgery. At both 6 months and 1 year after maze surgery, quality of life, measured by the SF-36, reached the levels of the general Swedish population. CONCLUSIONS: The maze operation can significantly improve the health-related quality of life in selected groups of patients with both paroxysmal and chronic AF refractory to antiarrhythmic therapy. PMID- 10840013 TI - Angiotensin II antagonist prevents electrical remodeling in atrial fibrillation. AB - BACKGROUND: The blockade of angiotensin II (Ang II) formation has protective effects on cardiovascular tissue; however, the role of Ang II in atrial electrical remodeling is unknown. The purpose of this study was to investigate the effects of candesartan and captopril on atrial electrical remodeling. METHODS AND RESULTS: In 24 dogs, the atrial effective refractory period (AERP) was measured before, during, and after rapid atrial pacing. Rapid atrial pacing at 800 bpm was maintained for 180 minutes. The infusion of saline (n=8), candesartan (n=5), captopril (n=6), or Ang II (n=5) was initiated 30 minutes before rapid pacing and continued throughout the study. In the saline group, AERP was significantly shortened during rapid atrial pacing (from 149+/-11 to 132+/-16 ms, P<0.01). There was no significant difference in AERP shortening between the saline group and the Ang II group. However, in the candesartan and captopril groups, shortening of the AERP after rapid pacing was completely inhibited (from 142+/-9 to 147+/-12 ms with candesartan, from 153+/-15 to 153+/-14 ms with captopril, P=NS). Although rate adaptation of the AERP was lost in the saline group, this phenomenon was preserved in the candesartan and captopril groups. CONCLUSIONS: The inhibition of endogenous Ang II prevented AERP shortening during rapid atrial pacing. These results indicate for the first time that Ang II may be involved in the mechanism of atrial electrical remodeling and that the blockade of Ang II may lead to the better therapeutic management of human atrial fibrillation. PMID- 10840014 TI - High glucose-induced apoptosis in human endothelial cells is mediated by sequential activations of c-Jun NH(2)-terminal kinase and caspase-3. AB - BACKGROUND: Diabetes mellitus causes multiple cardiovascular complications. High glucose can induce reactive oxygen species and apoptosis in endothelial cells. Little is known about the molecular mechanisms in high glucose-induced endothelial cell apoptosis. METHODS AND RESULTS: We elucidated the signaling pathway of high glucose-induced apoptosis in human umbilical vein endothelial cells (HUVECs). HUVECs were treated with media containing 5.5, 19, or 33 mmol/L of glucose in the presence or absence of an antioxidant, ascorbic acid. The level of intracellular H(2)O(2) was measured by flow cytometry. For detection of apoptosis, the cell death detection ELISA assay and the morphological Hoechst staining were used. High glucose was capable of inducing the activity of c-Jun NH(2)-terminal kinase (JNK) but not extracellular signal-regulated kinase 1/2 or p38 mitogen-activated protein kinase during the treatment periods, as evidenced by immunocomplex kinase assay. Moreover, we found that the interleukin 1beta converting enzyme (ICE)/CED-3 family protease (caspase-3) became activated in high glucose-induced apoptosis. Caspase-3/CPP32-specific inhibitor, Ac-DEVD-CHO, could inhibit high glucose-induced apoptosis. Furthermore, we found that JNK1 specific antisense oligonucleotide could suppress caspase-3 activity but not affect H(2)O(2) generation and could block apoptosis induced by high glucose. Also, H(2)O(2) generation, JNK activity, caspase-3 activity, and the subsequent apoptosis induced by high glucose could be suppressed by ascorbic acid. CONCLUSIONS: The present study indicates that reactive oxygen species induced by high glucose may be involved in JNK activation, which in turn triggers the caspase-3 that facilitates the apoptosis in HUVECs. PMID- 10840015 TI - Rapid electrical stimulation of contraction reduces the density of beta adrenergic receptors and responsiveness of cultured neonatal rat cardiomyocytes. Possible involvement of microtubule disassembly secondary to mechanical stress. AB - BACKGROUND: Although tachycardia is commonly present in patients with congestive heart failure, its role in the development of congestive heart failure remains unclear. We studied the effect of rapid electrical stimulation of contraction on beta-adrenergic receptor (beta-AR) signal pathway in cultured cardiomyocytes of neonatal rats. METHODS AND RESULTS: Contraction of cardiomyocytes was induced by electrical stimulation at 50 V with twice the threshold pulse width. beta-ARs were identified by [(3)H]CGP-12177 and [(3)H]dihydroalprenolol. Electrical stimulation reduced cell-surface but not total beta-AR density; the effect was dependent on pacing frequency (a reduction of 11%, 28%, and 18% in cells paced at 2.5, 3. 0, and 3.3 Hz, respectively). This reduction was apparent at 3 hours, in contrast to reduced beta-AR density after exposure to isoproterenol (ISP) for 1 hour. The fraction and inhibition constant of beta-AR binding agonist with high affinity were not affected by rapid electrical stimulation. In cardiomyocytes paced at 3.0 Hz for 24 hours, the response to ISP decreased compared with unpaced cells, 142% versus 204% of baseline with 1 micromol/L ISP, whereas the responses to forskolin or acetylcholine were not different. Treatment of cardiomyocytes with 2,3-butanedione monoxime (10 mmol/L) or taxol (10 micromol/L) inhibited the rapid pacing-induced reduction in beta-AR density. CONCLUSIONS: Our results suggest that contractile activity is involved in regulation of cardiac function by modulating the beta-AR system independently of hemodynamic and neurohormonal factors. This may help to elucidate the role of mechanical stress in the development of heart failure. PMID- 10840016 TI - Effects of experimental heart failure on atrial cellular and ionic electrophysiology. AB - BACKGROUND: Congestive heart failure (CHF) is frequently associated with atrial fibrillation (AF), but little is known about the effects of CHF on atrial cellular electrophysiology. METHODS AND RESULTS: We studied action potential (AP) properties and ionic currents in atrial myocytes from dogs with CHF induced by ventricular pacing at 220 to 240 bpm for 5 weeks. Atrial myocytes from CHF dogs were hypertrophied (mean+/-SEM capacitance, 89+/-2 pF versus 71+/-2 pF in control, n=160 cells per group, P<0.001). CHF significantly reduced the density of L-type Ca(2+) current (I(Ca)) by approximately 30%, of transient outward K(+) current (I(to)) by approximately 50%, and of slow delayed rectifier current (I(Ks)) by approximately 30% without altering their voltage dependencies or kinetics. The inward rectifier, ultrarapid and rapid delayed rectifier, and T type Ca(2+) currents were not altered by CHF. CHF increased transient inward Na(+)/Ca(2+) exchanger (NCX) current by approximately 45%. The AP duration of atrial myocytes was not altered by CHF at slow rates but was increased at faster rates, paralleling in vivo refractory changes. CHF created a substrate for AF, prolonging mean AF duration from 8+/-4 to 535+/-82 seconds (P<0.01). CONCLUSIONS: Experimental CHF selectively decreases atrial I(to), I(Ca), and I(Ks), increases NCX current, and leaves other currents unchanged. The cellular electrophysiological remodeling caused by CHF is quite distinct from that caused by atrial tachycardia, highlighting important differences in the cellular milieu characterizing different clinically relevant AF substrates. PMID- 10840017 TI - Calcium-activated Cl(-) current contributes to delayed afterdepolarizations in single Purkinje and ventricular myocytes. AB - BACKGROUND: The ionic mechanism underlying the transient inward current (I(ti)), the current responsible for delayed afterdepolarizations (DADs), appears to be different in ventricular myocytes and Purkinje fibers. In ventricular myocytes, I(ti) was ascribed to a Na(+)-Ca(2+) exchange current, whereas in Purkinje fibers, it was additionally ascribed to a Cl(-) current and a nonselective cation current. If Cl(-) current contributes to I(ti) and thus to DADs, Cl(-) current blockade may be potentially antiarrhythmogenic. In this study, we investigated the ionic nature of I(ti) in single sheep Purkinje and ventricular myocytes and the effects of Cl(-) current blockade on DADs. METHODS AND RESULTS: In whole-cell patch-clamp experiments, I(ti) was induced by repetitive depolarizations from -93 to +37 mV in the presence of 1 micromol/L norepinephrine. In both Purkinje and ventricular myocytes, I(ti) was inward at negative potentials and outward at positive potentials. The anion blocker 4,4'-diisothiocyanatostilbene-2,2' disulfonic acid (DIDS) blocked outward I(ti) completely but inward I(ti) only slightly. The DIDS-sensitive component of I(ti) was outwardly rectifying, with a reversal close to the reversal potential of Cl(-) currents. Blockade of Na(+) Ca(2+) exchange by substitution of extracellular Na(+) by equimolar Li(+) abolished the DIDS-insensitive component of I(ti). DIDS reduced both DAD amplitude and triggered activity based on DADs. Conclusions-In both Purkinje and ventricular myocytes, I(ti) consists of 2 ionic mechanisms: a Cl(-) current and a Na(+)-Ca(2+) exchange current. Blockade of the Cl(-) current may be potentially antiarrhythmogenic by lowering DAD amplitude and triggered activity based on DADs. PMID- 10840018 TI - Time course and mechanism of myocardial catecholamine release during transient ischemia in vivo. AB - BACKGROUND: Elevated concentrations of norepinephrine (NE) have been observed in ischemic myocardium. We investigated the magnitude and mechanism of catecholamine release in the myocardial interstitial fluid (MIF) during ischemia and reperfusion in vivo through the use of microdialysis. METHODS AND RESULTS: In 9 anesthetized pigs, interstitial catecholamine concentrations were measured in the perfusion areas of the left anterior descending coronary artery (LAD) and the left circumflex coronary artery. After stabilization, the LAD was occluded for 60 minutes and reperfused for 150 minutes. During the final 30 minutes, tyramine (154 nmol. kg(-1). min(-1)) was infused into the LAD. During LAD occlusion, MIF NE concentrations in the ischemic region increased progressively from 1. 0+/-0.1 to 524+/-125 nmol/L. MIF concentrations of dopamine and epinephrine rose from 0.4+/-0.1 to 43.9+/-9.5 nmol/L and from <0.2 (detection limit) to 4.7+/-0.7 nmol/L, respectively. Local uptake-1 blockade attenuated release of all 3 catecholamines by >50%. During reperfusion, MIF catecholamine concentrations returned to baseline within 120 minutes. At that time, the tyramine-induced NE release was similar to that seen in nonischemic control animals despite massive infarction. Arterial and MIF catecholamine concentrations in the left circumflex coronary artery region remained unchanged. CONCLUSIONS: Myocardial ischemia is associated with a pronounced increase of MIF catecholamines, which is at least in part mediated by a reversed neuronal reuptake mechanism. The increase of MIF epinephrine implies a (probably neuronal) cardiac source, whereas the preserved catecholamine response to tyramine in postischemic necrotic myocardium indicates functional integrity of sympathetic nerve terminals. PMID- 10840019 TI - Tissue factor overexpression in rat arterial neointima models thrombosis and progression of advanced atherosclerosis. AB - BACKGROUND: Tissue factor located in the atherosclerotic plaque might cause the clinically significant thrombotic events associated with end-stage disease. It might also affect intimal area by increasing matrix accumulation and stimulating smooth muscle cell (SMC) migration and proliferation. To test this hypothesis, we overexpressed tissue factor in a rat model of the human fibrous plaque. METHODS AND RESULTS: A neointima was generated by seeding tissue factor-overexpressing rat SMCs onto the luminal surface of a balloon-injured syngeneic rat carotid artery. The cells attached and expressed tissue factor over the long term. Mural thrombus accumulation was present at 4 and 7 days and increased neointimal SMC numbers and area by 2-fold at 2 and 4 weeks. Tissue factor overexpression accelerated reendothelialization compared with controls at 2 weeks and 1 month. Tissue factor-overexpressing SMCs exhibited increased migration both in vitro and in vivo. The increased migration by tissue factor-overexpressing SMCs in vitro was not dependent on activation of the coagulation cascade and could be blocked by an inhibitor of tissue factor. CONCLUSIONS: These results suggest that tissue factor plays a direct role in neointimal development by coagulation-dependent and -independent pathways. PMID- 10840020 TI - Orally administered unfractionated heparin with carrier agent is therapeutic for deep venous thrombosis. AB - BACKGROUND: Orally administered heparin (OHEP) is unreliable because of poor absorption. Sodium N-(8[2-hydroxybenzoyl]amino) caprylate (SNAC) is an amido acid that facilitates the gastrointestinal absorption of heparin. We evaluated the effectiveness of OHEP combined with SNAC (OHEP/SNAC) in the treatment of deep vein thrombosis (DVT). METHODS AND RESULTS: An internal jugular DVT was produced in 54 male Sprague-Dawley rats. Animals were assigned to 6 different groups for 7 days of treatment: untreated control, subcutaneous heparin (SC HEP) (300 U/kg SC TID), SNAC only (300 mg/kg PO TID), OHEP only (30 mg/kg PO TID), low-molecular weight heparin (LMWH) (enoxaparin 5 mg/kg SC QD), and OHEP/SNAC (30 mg/kg:300 mg/kg PO TID). The activated partial thromboplastin time (aPTT) and anti-factor X (anti-Xa) levels were measured. The incidence of residual DVT after 1 week of treatment was 100% (9 of 9) in the control group versus 10% (1 of 10) in the OHEP/SNAC and 10% (1 of 10) in the LMWH groups (P<0.001). There was also a significant reduction in clot weights between these groups. Compared with controls, there were no significant differences in the residual DVT in the SNAC only (6 of 6), OHEP-only (9 of 9), or SC HEP (8 of 10) groups. Combination OHEP/SNAC was as effective in the resolution of the clot and reducing clot weight as LMWH. The aPTT levels in the OHEP/SNAC group peaked at 30 minutes and were significantly higher than in all other groups (P<0.01). Anti-Xa levels were elevated at 15 minutes after dosing in the OHEP/SNAC group and remained significantly elevated at 4 hours (P<0.001). CONCLUSIONS: OHEP combined with a novel carrier agent (SNAC) successfully treated DVT in this rat model. PMID- 10840021 TI - Luigi Luciani and the earliest graphic demonstration of Wenckebach periodicity. AB - Using an isolated frog heart preparation with ligatures around the atria, Luigi Luciani, an Italian physiologist working in 1873 in Carl Ludwig's famous laboratory in Leipzig, was the first to demonstrate cardiac group beating, which he named periodic rhythm. He attributed this to increased resistance to impulse propagation between the atria and the ventricle. Karel F. Wenckebach, in his 1899 landmark report of group beating in a patient in which he also used pulse tracings, credited Luciani with this discovery. Wenckebach referred to the phenomena as "Luciani periods." With the advent of electrocardiography in the early 20th century, this form of group beating became known as Wenckebach periodicity and then as Mobitz type I atrioventricular block. We reanalyzed Luciani's original paper and pulse tracings, and we show that periodic rhythm does indeed meet the criteria of second-degree atrioventricular block as established by Wenckebach. We also reviewed the career of Luciani, who was an important investigator, outstanding teacher and mentor, and distinguished leader of 19th-century physiology. We conclude that Wenckebach still deserves to have his name eponymously attached to this type of atrioventricular block because he was the first to unravel the complicated relationship between atrial and ventricular conduction. PMID- 10840022 TI - Heartbreaking roadwork. PMID- 10840023 TI - Mental versus physical stress, QT prolongation, and the autonomic nervous system. PMID- 10840024 TI - Meeting highlights: highlights of the 72nd Scientific Sessions of the American Heart Association. PMID- 10840025 TI - Numbers of uninsured increases. PMID- 10840026 TI - Medicare prescription drug battle heats up. PMID- 10840027 TI - Activation of heterotrimeric G-protein signaling by a ras-related protein. Implications for signal integration. AB - Utilizing a functional screen in the yeast Saccharomyces cerevisiae we identified mammalian proteins that activate heterotrimeric G-protein signaling pathways in a receptor-independent fashion. One of the identified activators, termed AGS1 (for activator of G-protein signaling), is a human Ras-related G-protein that defines a distinct subgroup of the Ras superfamily. Expression of AGS1 in yeast and in mammalian cells results in specific activation of Galpha(i)/Galpha(o) heterotrimeric signaling pathways. In addition, the in vivo and in vitro properties of AGS1 are consistent with it functioning as a direct guanine nucleotide exchange factor for Galpha(i)/Galpha(o). AGS1 thus presents a unique mechanism for signal integration via heterotrimeric G-protein signaling pathways. PMID- 10840028 TI - Plant chimeric Ca2+/Calmodulin-dependent protein kinase. Role of the neural visinin-like domain in regulating autophosphorylation and calmodulin affinity. AB - Chimeric Ca(2+)/calmodulin-dependent protein kinase (CCaMK) is characterized by a serine-threonine kinase domain, an autoinhibitory domain, a calmodulin-binding domain and a neural visinin-like domain with three EF-hands. The neural visinin like Ca(2+)-binding domain at the C-terminal end of the CaM-binding domain makes CCaMK unique among all the known calmodulin-dependent kinases. Biological functions of the plant visinin-like proteins or visinin-like domains in plant proteins are not well known. Using EF-hand deletions in the visinin-like domain, we found that the visinin-like domain regulated Ca(2+)-stimulated autophosphorylation of CCaMK. To investigate the effects of Ca(2+)-stimulated autophosphorylation on the interaction with calmodulin, the equilibrium binding constants of CCaMK were measured by fluorescence emission anisotropy using dansylated calmodulin. Binding was 8-fold tighter after Ca(2+)-stimulated autophosphorylation. This shift in affinity did not occur in CCaMK deletion mutants lacking Ca(2+)-stimulated autophosphorylation. A variable calmodulin affinity regulated by Ca(2+)-stimulated autophosphorylation mediated through the visinin-like domain is a new regulatory mechanism for CCaMK activation and calmodulin-dependent protein kinases. Our experiments demonstrate the existence of two functional molecular switches in a protein kinase regulating the kinase activity, namely a visinin-like domain acting as a Ca(2+)-triggered switch and a CaM-binding domain acting as an autophosphorylation-triggered molecular switch. PMID- 10840029 TI - Apoptosis induced by the nuclear death domain protein p84N5 is associated with caspase-6 and NF-kappa B activation. AB - Although the mechanisms involved in responses to extracellular or mitochondrial apoptotic signals have received considerable attention, the mechanisms utilized within the nucleus to transduce apoptotic signals are not well understood. We have characterized apoptosis induced by the nuclear death domain-containing protein p84N5. Adenovirus-mediated N5 gene transfer or transfection of p84N5 expression vectors induces apoptosis in tumor cell lines with nearly 100% efficiency as indicated by cellular morphology, DNA fragmentation, and annexin V staining. Using peptide substrates and Western blotting, we have determined that N5-induced apoptosis is initially accompanied by activation of caspase-6. Activation of caspases-3 and -9 does not peak until 3 days after the peak of caspase-6 activity. Expression of p84N5 also leads to activation of NF-kappaB as indicated by nuclear translocation of p65RelA and transcriptional activation of a NF-kappaB-dependent reporter promoter. Changes in the relative expression level of Bcl-2 family proteins, including Bak and Bcl-Xs, are also observed during p84N5-induced apoptosis. Finally, we demonstrate that p84N5-induced apoptosis does not require p53 and is not inhibited by p53 coexpression. We propose that p84N5 is involved in an apoptotic pathway distinct from those triggered by death domain-containing receptors or by p53. PMID- 10840030 TI - Detection of a novel quiescence-dependent protein kinase. AB - We have identified a cell quiescence-specific 33-kDa cytoplasmic protein kinase (p33(QIK), Quiescence-Induced Kinase) based on induction of p33(QIK)-specific kinase activity of cells growth-arrested in the quiescent phase and deactivation upon entry into the cell cycle. Blockage of macromolecular synthesis prevents p33(QIK) from deactivation, indicating a requirement of newly synthesized regulators for deactivation of p33(QIK) during G(0)/G(1) transition. Stress shock induces additional increases of p33(QIK) activity in a quiescence-dependent manner that correlates with induction of apoptosis. Using a specific antibody to Krs1/Mst2 protein, we found that p33(QIK) is related to p63(Krs1) and is distinguishable from a 36-kDa protein kinase, which is induced through proteolytic modification of activated p63(Krs1) in proliferating cells undergoing apoptosis. p33(QIK) is constantly expressed in quiescent, proliferating, and apoptotic quiescent cells. Regulation of p33(QIK) activity involves protein phosphorylation/dephosphorylation in a proteolysis-independent manner. Regulation of p33(QIK) and related p63(Krs1) and p36 appears to involve distinct pathways in quiescent and proliferating cells, respectively. Our results illustrate the relevance of p33(QIK) activity for cell quiescence that may provide a new insight into signaling pathways regulated in cells during quiescence and quiescence related apoptosis. PMID- 10840031 TI - Complexes of the G protein subunit gbeta 5 with the regulators of G protein signaling RGS7 and RGS9. Characterization in native tissues and in transfected cells. AB - A novel protein class, termed regulators of G protein signaling (RGS), negatively regulates G protein pathways through a direct interaction with Galpha subunits and stimulation of GTP hydrolysis. An RGS subfamily including RGS6, -7, -9, and 11, which contain a characteristic Ggamma -like domain, also has the unique ability to interact with the G protein beta subunit Gbeta(5). Here, we examined the behavior of Gbeta(5), RGS7, RGS9, and Galpha in tissue extracts using immunoprecipitation and conventional chromatography. Native Gbeta(5) and RGS7 from brain, as well as photoreceptor-specific Gbeta(5)L and RGS9, always co purified as tightly associated dimers, and neither RGS-free Gbeta(5) nor Gbeta(5) free RGS could be detected. Co-expression in COS-7 cells of Gbeta(5) dramatically increased the protein level of RGS7 and vice versa, indicating that cells maintain Gbeta(5):RGS stoichiometry in a manner similar to Gbetagamma complexes. This mechanism is non-transcriptional and is based on increased protein stability upon dimerization. Thus, analysis of native Gbeta(5)-RGS and their coupled expression argue that in vivo, Gbeta(5) and Ggamma-like domain-containing RGSs only exist as heterodimers. Native Gbeta(5)-RGS7 did not co-precipitate or co purify with Galpha(o) or Galpha(q); nor did Gbeta(5)L-RGS9 with Galpha(t). However, in transfected cells, RGS7 and Gbeta(5)-RGS7 inhibited Galpha(q) mediated Ca(2+) response to muscarinic M3 receptor activation. Thus, Gbeta(5)-RGS dimers differ from other RGS proteins in that they do not bind to Galpha with high affinity, but they can still inhibit G protein signaling. PMID- 10840032 TI - Characterization and gene expression of high conductance calcium-activated potassium channels displaying mechanosensitivity in human odontoblasts. AB - Odontoblasts form a layer of cells responsible for the dentin formation and possibly mediate early stages of sensory processing in teeth. Several classes of ion channels have previously been identified in the odontoblast or pulp cell membrane, and it is suspected that these channels assist in these events. This study was carried out to characterize the K(Ca) channels on odontoblasts fully differentiated in vitro using the patch clamp technique and to investigate the HSLO gene expression encoding the alpha-subunit of these channels on odontoblasts in vivo. In inside-out patches, K(Ca) channels were identified on the basis of their K(+) selectivity, conductance, voltage, and Ca(2+) dependence. In cell attached patches, these channels were found to be activated by application of a negative pressure as well as an osmotic shock. By reverse transcription polymerase chain reaction, a probe complementary to K(Ca) alpha-subunit mRNA was constructed and used for in situ hybridization on human dental pulp samples. Transcripts were expressed in the odontoblast layer. The use of antibodies showed that the K(Ca) channels were preferentially detected at the apical pole of the odontoblasts. These channels could be involved in mineralization processes. Their mechanosensitivity suggests that the fluid displacement within dentinal tubules could be transduced into electrical cell signals. PMID- 10840033 TI - Mutations in the estrogen receptor ligand binding domain discriminate between hormone-dependent transactivation and transrepression. AB - The estrogen receptor (ER) suppresses transcriptional activity of the RelA subunit of nuclear factor-kappaB in a hormone-dependent manner by a mechanism involving both the receptor DNA binding domain and ligand binding domain (LBD). In this study we examine the role of the ER LBD in mediating ligand-dependent RelA transrepression. Both ERalpha and ERbeta inhibit RelA in response to 17beta estradiol but not in the presence of antihormones. We have identified residues within the ERalpha LBD that are responsible for receptor dimerization and show that dimerization is necessary for transactivation and transrepression. Moreover we have generated mutant receptors that have lost their ability to inhibit RelA but retain their capacity to stimulate transcription and conversely mutants that are transcriptionally defective but capable of antagonizing RelA. Overexpression of p160 and cAMP-response element-binding protein-binding protein/p300 co activators failed to relieve repression of RelA, which is consistent with the demonstration that RelA inhibition can occur independently of these co activators. These findings suggest it is unlikely that sequestration of these cofactors required for ER transcriptional activation can account for hormone dependent antagonism of RelA. The identification of ER mutants that discriminate between transactivation and transrepression implies that distinct surfaces within the LBD are involved in mediating these two receptor functions. PMID- 10840034 TI - The Rho family GTPase Cdc42 regulates the activation of Ras/MAP kinase by the exchange factor Ras-GRF. AB - The Ras guanine-nucleotide exchange factor Ras-GRF/Cdc25(Mn) harbors a complex array of structural motifs that include a Dbl-homology (DH) domain, usually found in proteins that interact functionally with the Rho family GTPases, and the role of which is not yet fully understood. Here, we present evidence that Ras-GRF requires its DH domain to translocate to the membrane, to stimulate exchange on Ras, and to activate mitogen-activated protein kinase (MAPK). In an unprecedented fashion, we have found that these processes are regulated by the Rho family GTPase Cdc42. We show that GDP- but not GTP-bound Cdc42 prevents Ras-GRF recruitment to the membrane and activation of Ras/MAPK, although no direct association of Ras-GRF with Cdc42 was detected. We also demonstrate that catalyzing GDP/GTP exchange on Cdc42 facilitates Ras-GRF-induced MAPK activation. Moreover, we show that the potentiating effect of ionomycin on Ras-GRF-mediated MAPK stimulation is also regulated by Cdc42. These results provide the first evidence for the involvement of a Rho family G protein in the control of the activity of a Ras exchange factor. PMID- 10840035 TI - beta 2-adrenergic receptor activates extracellular signal-regulated kinases (ERKs) via the small G protein rap1 and the serine/threonine kinase B-Raf. AB - G protein-coupled receptors can induce cellular proliferation by stimulating the mitogen-activated protein (MAP) kinase cascade. Heterotrimeric G proteins are composed of both alpha and betagamma subunits that can signal independently to diverse intracellular signaling pathways including those that activate MAP kinases. In this study, we examined the ability of isoproterenol, an agonist of the beta(2)-adrenergic receptor (beta(2)AR), to stimulate extracellular signal regulated kinases (ERKs). Using HEK293 cells, which express endogenous beta(2)AR, we show that isoproterenol stimulates ERKs via beta(2)AR. This action of isoproterenol requires cAMP-dependent protein kinase and is insensitive to pertussis toxin, suggesting that Galpha(s) activation of cAMP-dependent protein kinase is required. Interestingly, beta(2)AR activates both the small G proteins Rap1 and Ras, but only Rap1 is capable of coupling to Raf isoforms. beta(2)AR inhibits the Ras-dependent activation of both Raf isoforms Raf-1 and B-Raf, whereas Rap1 activation by isoproterenol recruits and activates B-Raf. beta(2)AR activation of ERKs is not blocked by expression of RasN17, an interfering mutant of Ras, but is blocked by expression of either RapN17 or Rap1GAP1, both of which interfere with Rap1 signaling. We propose that isoproterenol can activate ERKs via Rap1 and B-Raf in these cells. PMID- 10840036 TI - Identification and substrate specificity of beta -ketoacyl (acyl carrier protein) synthase III (mtFabH) from Mycobacterium tuberculosis. AB - The long-chain alpha-alkyl-beta-hydroxy fatty acids, termed mycolic acids, which are characteristic components of the mycobacterial cell wall are produced by successive rounds of elongation catalyzed by a multifunctional (type I) fatty acid synthase complex followed by a dissociated (type II) fatty acid synthase. In bacterial type II systems, the first initiation step in elongation is the condensation of acetyl-CoA with malonyl-acyl carrier protein (ACP) catalyzed by beta-ketoacyl-ACP III (FabH). An open reading frame in the Mycobacterium tuberculosis genome (Rv0533c), now termed mtfabH, was 37.3% identical to Escherichia coli ecFabH and contained the Cys-His-Asn catalytic triad signature. However, the purified recombinant mtFabH clearly preferred long-chain acyl-CoA substrates rather than acyl-ACP primers and did not utilize acetyl-CoA as a primer in comparison to ecFabH. In addition, purified mtFabH was sensitive to thiolactomycin and resistant to cerulenin in an in vitro assay. However, mtFabH overexpression in Mycobacterium bovis BCG did not confer thiolactomycin resistance, suggesting that mtFabH may not be the primary target of thiolactomycin inhibition in vivo and led to several changes in the lipid composition of the bacilli. The data presented is consistent with a role for mtFabH as the pivotal link between the type I and type II fatty acid elongation systems in M. tuberculosis. This study opens up new avenues for the development of selective and novel anti-mycobacterial agents targeted against mtFabH. PMID- 10840037 TI - Importance of protein kinase C targeting for the phosphorylation of its substrate, myristoylated alanine-rich C-kinase substrate. AB - We visualized the translocation of myristoylated alanine-rich protein kinase C substrate (MARCKS) in living Chinese hamster ovary-K1 cells using MARCKS tagged to green fluorescent protein (MARCKS-GFP). MARCKS-GFP was rapidly translocated from the plasma membrane to the cytoplasm after the treatment with phorbol ester, which translocates protein kinase C (PKC) to the plasma membrane. In contrast, PKC activation by hydrogen peroxide, which was not accompanied by PKC translocation, did not alter the intracellular localization of MARCKS-GFP. Non myristoylated mutant of MARCKS-GFP was distributed throughout the cytoplasm, including the nucleoplasm, and was not translocated by phorbol ester or by hydrogen peroxide. Phosphorylation of wild-type MARCKS-GFP was observed in cells treated with phorbol ester but not with hydrogen peroxide, whereas non myristoylated mutant of MARCKS-GFP was phosphorylated in cells treated with hydrogen peroxide but not with phorbol ester. Phosphorylation of both MARCKS-GFPs reduced the amount of F-actin. These findings revealed that PKC targeting to the plasma membrane is required for the phosphorylation of membrane-associated MARCKS and that a mutant MARCKS existing in the cytoplasm can be phosphorylated by PKC activated in the cytoplasm without translocation but not by PKC targeted to the membrane. PMID- 10840038 TI - 14-3-3zeta is an effector of tau protein phosphorylation. AB - Neurofibrillary tangles associated with Alzheimer's disease are composed mainly of paired helical filaments that are formed by the aggregation of abnormally phosphorylated microtubule-associated protein tau. 14-3-3, a highly conserved protein family that exists as seven isoforms and regulates diverse cellular processes is present in neurofibrillary tangles (Layfield, R., Fergusson, J., Aitken, A., Lowe, J., Landon, M., Mayer, R. J. (1996) Neurosci. Lett. 209, 57 60). The role of 14-3-3 in Alzheimer's disease pathogenesis is not known. In this study, we found that the 14-3-3zeta isoform is associated with tau in brain extract and profoundly stimulates cAMP-dependent protein kinase catalyzed in vitro phosphorylation on Ser(262)/Ser(356) located within the microtubule-binding region of tau. 14-3-3zeta binds to both phosphorylated and nonphosphorylated tau, and the binding site is located within the microtubule-binding region of tau. From brain extract, 14-3-3zeta co-purifies with microtubules, and tubulin blocks 14-3-3zeta-tau binding. Among four 14-3-3 isoforms tested, beta and zeta but not gamma and epsilon associate with tau. Our data suggest that 14-3-3zeta is a tau protein effector and may be involved in the abnormal tau phosphorylation occurring during Alzheimer's disease ontogeny. PMID- 10840040 TI - Paxillin localizes to the lymphocyte microtubule organizing center and associates with the microtubule cytoskeleton. AB - Paxillin is a focal adhesion-associated protein that functions as a multi-domain adapter protein, binding several structural and signaling molecules. alpha Tubulin was identified as an interacting protein in a two-hybrid screen using the paxillin C-terminal LIM domain as a bait. In vitro binding assays with glutathione S-transferase-paxillin demonstrated an interaction of alpha-tubulin with the C terminus of paxillin. Another member of the tubulin family, gamma tubulin, bound to both the N and the C terminus of paxillin. The interaction between paxillin and both alpha- and gamma-tubulin in vivo was confirmed by co immunoprecipitation from human T lymphoblasts. Immunofluorescence studies revealed that, in adherent T cells, paxillin localized to sites of cell-matrix interaction as well as to a large perinuclear region. Confocal microscopy revealed that this region corresponds to the lymphocyte microtubule organizing center, where paxillin colocalizes with alpha- and gamma-tubulin. The localization of paxillin to this area was observed in cells in suspension as well as during adhesion to integrin ligands. These data constitute the first characterization of the interaction of paxillin with the microtubule cytoskeleton, and suggest that paxillin, in addition to its well established role at focal adhesions, could also be associated with the lymphocyte microtubule network. PMID- 10840041 TI - Functional analysis of tail domains of Acanthamoeba myosin IC by characterization of truncation and deletion mutants. AB - Acanthamoeba myosin IC has a single 129-kDa heavy chain and a single 17-kDa light chain. The heavy chain comprises a 75-kDa catalytic head domain with an ATP sensitive F-actin-binding site, a 3-kDa neck domain, which binds a single 17-kDa light chain, and a 50-kDa tail domain, which binds F-actin in the presence or absence of ATP. The actin-activated MgATPase activity of myosin IC exhibits triphasic actin dependence, apparently as a consequence of the two actin-binding sites, and is regulated by phosphorylation of Ser-329 in the head. The 50-kDa tail consists of a basic domain, a glycine/proline/alanine-rich (GPA) domain, and a Src homology 3 (SH3) domain, often referred to as tail homology (TH)-1, -2, and -3 domains, respectively. The SH3 domain divides the TH-3 domain into GPA-1 and GPA-2. To define the functions of the tail domains more precisely, we determined the properties of expressed wild type and six mutant myosins, an SH3 deletion mutant and five mutants truncated at the C terminus of the SH3, GPA-2, TH-1, neck and head domains, respectively. We found that both the TH-1 and GPA-2 domains bind F-actin in the presence of ATP. Only the mutants that retained an actin binding site in the tail exhibited triphasic actin-dependent MgATPase activity, in agreement with the F-actin-cross-linking model, but truncation reduced the MgATPase activity at both low and high actin concentrations. Deletion of the SH3 domain had no effect. Also, none of the tail domains, including the SH3 domain, affected either the K(m) or V(max) for the phosphorylation of Ser-329 by myosin I heavy chain kinase. PMID- 10840042 TI - Superagonistic activation of ErbB-1 by EGF-related growth factors with enhanced association and dissociation rate constants. AB - Epidermal growth factor (EGF) and transforming growth factor-alpha (TGFalpha) are mitogenic hormones that exert their activity primarily by binding to the EGF receptor, also known as ErbB-1. We have recently characterized a set of EGF/TGFalpha chimeric molecules with similar high affinity for ErbB-1 as EGF and TGFalpha and shown that three of these chimeras induce mitogenic cell stimulation at already a 10-fold lower concentration than their wild-type counterparts (Lenferink, A. E., Kramer, R. H., van Vugt, M. J., Konigswieser, M., DiFiore, P. P., van Zoelen, E. J., and van de Poll, M. L. (1997) Biochem. J. 327, 859-865). In the present study we show that these so-called superagonistic chimeras do not differ from EGF and TGFalpha in their ability to induce ErbB-1 tyrosine phosphorylation but are considerably more potent in activation of mitogen activated protein kinase phosphorylation. Direct cell binding studies and analysis of ligand-receptor interaction by surface plasmon resonance measurements revealed that both the association rate constant (k(on)) and the dissociation rate constant (k(off)) of these superagonists is 3-5-fold higher in comparison with the wild-type ligands and nonsuperagonistic chimeras. These data indicate that the dynamic on and off rate constants for receptor binding may be more specific parameters for determining the mitogenic activity of peptide hormones than their constants for equilibrium receptor binding. PMID- 10840043 TI - Structural evidence for ligand specificity in the binding domain of the human androgen receptor. Implications for pathogenic gene mutations. AB - The crystal structures of the human androgen receptor (hAR) and human progesterone receptor ligand-binding domains in complex with the same ligand metribolone (R1881) have been determined. Both three-dimensional structures show the typical nuclear receptor fold. The change of two residues in the ligand binding pocket between the human progesterone receptor and hAR is most likely the source for the specificity of R1881 to the hAR. The structural implications of the 14 known mutations in the ligand-binding pocket of the hAR ligand-binding domains associated with either prostate cancer or the partial or complete androgen receptor insensitivity syndrome were analyzed. The effects of most of these mutants could be explained on the basis of the crystal structure. PMID- 10840044 TI - Sequestration of the active site by interdomain shifting. Crystallographic and spectroscopic evidence for distinct conformations of L-3-hydroxyacyl-CoA dehydrogenase. AB - l-3-Hydroxyacyl-CoA dehydrogenase reversibly catalyzes the conversion of l-3 hydroxyacyl-CoA to 3-ketoacyl-CoA concomitant with the reduction of NAD(+) to NADH as part of the beta-oxidation spiral. In this report, crystal structures have been solved for the apoenzyme, binary complexes of the enzyme with reduced cofactor or 3-hydroxybutyryl-CoA substrate, and an abortive ternary complex of the enzyme with NAD(+) and acetoacetyl-CoA. The models illustrate positioning of cofactor and substrate within the active site of the enzyme. Comparison of these structures with the previous model of the enzyme-NAD(+) complex reveals that although significant shifting of the NAD(+)-binding domain relative to the C terminal domain occurs in the ternary and substrate-bound complexes, there are few differences between the apoenzyme and cofactor-bound complexes. Analysis of these models clarifies the role of key amino acids implicated in catalysis and highlights additional critical residues. Furthermore, a novel charge transfer complex has been identified in the course of abortive ternary complex formation, and its characterization provides additional insight into aspects of the catalytic mechanism of l-3-hydroxyacyl-CoA dehydrogenase. PMID- 10840045 TI - Membrane-embedded synaptotagmin penetrates cis or trans target membranes and clusters via a novel mechanism. AB - The synaptic vesicle protein synaptotagmin I has been proposed to serve as a Ca(2+) sensor for rapid exocytosis. Synaptotagmin spans the vesicle membrane once and possesses a cytoplasmic domain largely comprised of two C2 domains designated C2A and C2B. We have determined how deep the Ca(2+)-binding loops of Ca(2+).C2A penetrate into the lipid bilayer and report mutations in synaptotagmin that can uncouple membrane penetration from Ca(2+)-triggered interactions with the SNARE complex. To determine whether C2A penetrates into the vesicle ("cis") or plasma ("trans") membrane, we reconstituted a fragment of synaptotagmin that includes the membrane-spanning and C2A domain (C2A-TMR) into proteoliposomes. Kinetics experiments revealed that cis interactions are rapid (< or =500 micros). Binding in the trans mode was distinguished by the slow diffusion of trans target vesicles. Both modes of binding were observed, indicating that the linker between the membrane anchor and C2A domain functions as a flexible tether. C2A-TMR assembled into oligomers via a novel N-terminal oligomerization domain suggesting that synaptotagmin may form clusters on the surface of synaptic vesicles. This novel mode of clustering may allow for rapid Ca(2+)-triggered oligomerization of the protein via the membrane distal C2B domain. PMID- 10840046 TI - Barnacle cement proteins. Importance of disulfide bonds in their insolubility. AB - Barnacles produce a cement that is a proteinaceous underwater adhesive for their secure attachment to the substratum. The biochemical properties of the cement have not previously been elucidated, because the insolubility of the cement proteins hampers their purification and characterization. We developed a non hydrolytic method to render soluble most of the cement components, thereby allowing the proteins to be analyzed. Megabalanus rosa cement could be almost completely rendered soluble by its reduction with 0.5 m dithiothreitol at 60 degrees C in a 7 m guanidine hydrochloride solution, the high concentration of dithiothreitol being indispensable to achieve this. The effectiveness of this reduction treatment was confirmed by the detachment of the barnacle from the substratum. Three proteins comprising up to 94% of the whole cement were identified as the major cement components. The cDNA clone of one of these major proteins was isolated, and the site-specific expression of the gene in the basal portion of the adult barnacle, where the cement glands are located, was demonstrated. A sequence analysis revealed this cement component to be a novel protein of 993 amino acid residues, including a signal peptide. This is the first report of the major component of the barnacle cement protein complex. PMID- 10840047 TI - Negative regulation of central nervous system myelination by polysialylated neural cell adhesion molecule. AB - Many factors have been shown to promote myelination, but few have been shown to be inhibitory. Here, we show that polysialylated-neural cell adhesion molecule (PSA-NCAM) can negatively regulate myelin formation. During development, PSA-NCAM is first expressed on all growing fibers; then, axonal expression is down regulated and myelin deposition occurs only on PSA-NCAM-negative axons. Similarly, in cocultures of oligodendrocytes and neurons, PSA-NCAM expression on axons is initially high, but decreases as myelination proceeds. Importantly, if expression of PSA-NCAM is prematurely decreased in cultures, by either antibody mediated internalization or enzymatic removal of the PSA moieties with endoneuraminidase N (endo-N), myelination increases 4- to 5-fold. In the optic nerve, premature cleavage of PSA moieties by intravitreous injection of endo-N also induces a transient increase in the number of myelinated internodes, but does not interfere with the onset of myelination. Previously, we showed that axonal electrical activity strongly induced myelination, which could be prevented by tetrodotoxin (TTX), an action potential blocker. Interestingly, removal of PSA moieties does not reverse the inhibition of myelination by TTX. Together, this suggests that myelination is tightly controlled by both positive (electrical activity) and negative (PSA-NCAM expression) regulatory signals. PMID- 10840049 TI - Do lampreys have lymphocytes? The Spi evidence. AB - It is generally accepted that living jawless vertebrates (lampreys and hagfishes) lack the capability of mounting an adaptive immune response. At the same time, however, there are reports describing histological evidence for the presence in agnathan tissues of lymphocytes, the key players in adaptive immunity. The question therefore arises whether the cells identified morphologically as lymphocytes are true lymphocytes in terms of their genetic developmental program. In this study, evidence is provided that the lampreys express a member of the purine box 1 (PU.1)/spleen focus-forming virus integration B (Spi-B) gene family known to be critically and specifically involved in the differentiation of lymphocytes in jawed vertebrates. The lamprey gene is expressed in the lymphocyte like cells of the digestive tract and inexplicably also in the ovary. PMID- 10840050 TI - Multidrug-resistance protein 5 is a multispecific organic anion transporter able to transport nucleotide analogs. AB - Two prominent members of the ATP-binding cassette superfamily of transmembrane proteins, multidrug resistance 1 (MDR1) P-glycoprotein and multidrug resistance protein 1 (MRP1), can mediate the cellular extrusion of xenobiotics and (anticancer) drugs from normal and tumor cells. The MRP subfamily consists of at least six members, and here we report the functional characterization of human MRP5. We found resistance against the thiopurine anticancer drugs, 6 mercaptopurine (6-MP) and thioguanine, and the anti-HIV drug 9-(2 phosphonylmethoxyethyl)adenine (PMEA) in MRP5-transfected cells. This resistance is due to an increased extrusion of PMEA and 6-thioinosine monophosphate from the cells that overproduce MRP5. In polarized Madin-Darby canine kidney II (MDCKII) cells transfected with an MRP5 cDNA construct, MRP5 is routed to the basolateral membrane and these cells transport S-(2,4-dinitrophenyl)glutathione and glutathione preferentially toward the basal compartment. Inhibitors of organic anion transport inhibit transport mediated by MRP5. We speculate that MRP5 might play a role in some cases of unexplained resistance to thiopurines in acute lymphoblastic leukemia and/or to antiretroviral nucleoside analogs in HIV infected patients. PMID- 10840051 TI - Multiple differences in gene expression in regulatory Valpha 24Jalpha Q T cells from identical twins discordant for type I diabetes. AB - Quantitative and qualitative defects in CD1d-restricted T cells have been demonstrated in human and murine autoimmune diseases. To investigate the transcriptional consequences of T cell receptor activation in human Valpha24JalphaQ T cell clones, DNA microarrays were used to quantitate changes in mRNA levels after anti-CD3 stimulation of clones derived from identical twins discordant for type 1 diabetes and IL-4 secretion. Activation resulted in significant modulation of 226 transcripts in the IL-4 secreting clone and 86 in the IL-4-null clone. Only 28 of these genes were in common. The differences observed suggest both ineffective differentiation of diabetic Valpha24JalphaQ T cells and a role for invariant T cells in the recruitment and activation of cells from the myeloid lineage. PMID- 10840052 TI - Stepping rotation of F1-ATPase visualized through angle-resolved single fluorophore imaging. AB - Orientation dependence of single-fluorophore intensity was exploited in order to videotape conformational changes in a protein machine in real time. The fluorophore Cy3 attached to the central subunit of F(1)-ATPase revealed that the subunit rotates in the molecule in discrete 120 degrees steps and that each step is driven by the hydrolysis of one ATP molecule. These results, unlike those from the previous study under a frictional load, show that the 120 degrees stepping is a genuine property of this molecular motor. The data also show that the rate of ATP binding is insensitive to the load exerted on the rotor subunit. PMID- 10840053 TI - Long-term enzyme correction and lipid reduction in multiple organs of primary and secondary transplanted Fabry mice receiving transduced bone marrow cells. AB - Fabry disease is a compelling target for gene therapy as a treatment strategy. A deficiency in the lysosomal hydrolase alpha-galactosidase A (alpha-gal A; EC ) leads to impaired catabolism of alpha-galactosyl-terminal lipids such as globotriaosylceramide (Gb3). Patients develop vascular occlusions that cause cardiovascular, cerebrovascular, and renal disease. Unlike for some lysosomal storage disorders, there is limited primary nervous system involvement in Fabry disease. The enzyme defect can be corrected by gene transfer. Overexpression of alpha-gal A by transduced cells results in secretion of this enzyme. Secreted enzyme is available for uptake by nontransduced cells presumably by receptor mediated endocytosis. Correction of bystander cells may occur locally or systemically after circulation of the enzyme in the blood. In this paper we report studies on long-term genetic correction in an alpha-gal A-deficient mouse model of Fabry disease. alpha-gal A-deficient bone marrow mononuclear cells (BMMCs) were transduced with a retrovirus encoding alpha-gal A and transplanted into sublethally and lethally irradiated alpha-gal A-deficient mice. alpha-gal A activity and Gb3 levels were analyzed in plasma, peripheral blood mononuclear cells, BMMCs, liver, spleen, heart, lung, kidney, and brain. Primary recipient animals were followed for up to 26 weeks. BMMCs were then transplanted into secondary recipients. Increased alpha-gal A activity and decreased Gb3 storage were observed in all recipient groups in all organs and tissues except the brain. These effects occurred even with a low percentage of transduced cells. The findings indicate that genetic correction of bone marrow cells derived from patients with Fabry disease may have utility for phenotypic correction of patients with this disorder. PMID- 10840054 TI - A downstream element in the human beta-globin promoter: evidence of extended sequence-specific transcription factor IID contacts. AB - We describe here the identification and characterization of a functional downstream element in the human adult beta-globin promoter. The existence of this element was indicated by two mutations at +22 and +33 downstream of the beta globin transcriptional start site in humans with beta-thalassemia. In vitro transcriptional analysis of these mutants, plus a third at +13, indicates that all three decrease transcription from the beta-globin promoter. Scanning mutagenesis from +10 to +45 indicates that this region contains a functional cis element(s) in vitro, and we designated this element the DCE (downstream core element). The DCE functions in concert with the beta-globin CATA box and initiator element, as well as in a heterologous, TATA-less context. A second set of mutants indicates that a particular geometry of the DCE and core promoter is necessary for promoter function. Lastly, DCE mutants show reduced affinity for transcription factor IID (TFIID). These data indicate that TFIID makes sequence specific contacts to the DCE and that TFIID binding is necessary for DCE function. PMID- 10840055 TI - Peripheral infection with adenovirus causes unexpected long-term brain inflammation in animals injected intracranially with first-generation, but not with high-capacity, adenovirus vectors: toward realistic long-term neurological gene therapy for chronic diseases. AB - Although adenoviral vectors provide prolonged gene expression in the brain by comparison to peripheral organs, expression is eliminated by a severe inflammatory infiltration (i.e., activated macrophages/microglia and T lymphocytes) after peripheral infection with adenovirus. Here, we demonstrate that high-capacity adenoviral (HC-Ad) vectors succeed in maintaining long-term transgene expression in the brain, even in the presence of an active peripheral immunization with adenovirus that completely eliminates expression from first generation vectors within 60 days. Importantly, even 60 days after the peripheral infection, brains injected with first-generation vectors exhibited evidence of a chronic infiltration of CD8(+) cells, macrophage/microglial activation, and up regulation of brain MHC-I expression. No inflammation was observed in the brains injected with the HC-Ad vector. Thus, these results demonstrate that HC-Ad vectors will allow safe, stable, and long-term transgene expression in the brain, even in the presence of peripheral infection with adenovirus. This markedly improves the prospects for the use of adenoviral vectors for long-term gene therapy of neurological disorders. PMID- 10840056 TI - Opiates inhibit neurogenesis in the adult rat hippocampus. AB - Recent work implicates regulation of neurogenesis as a form of plasticity in the adult rat hippocampus. Given the known effects of opiates such as morphine and heroin on hippocampal function, we examined opiate regulation of neurogenesis in this brain region. Chronic administration of morphine decreased neurogenesis by 42% in the adult rat hippocampal granule cell layer. A similar effect was seen in rats after chronic self-administration of heroin. Opiate regulation of neurogenesis was not mediated by changes in circulating levels of glucocorticoids, because similar effects were seen in rats that received adrenalectomy and corticosterone replacement. These findings suggest that opiate regulation of neurogenesis in the adult rat hippocampus may be one mechanism by which drug exposure influences hippocampal function. PMID- 10840057 TI - True optical resolution beyond the Rayleigh limit achieved by standing wave illumination. AB - During the last decade, various efforts have been undertaken to enhance the resolution of optical microscopes, mostly because of their importance in biological sciences. Herein, we describe a method to increase the resolution of fluorescence microscopy by illuminating the specimen with a mesh-like interference pattern of a laser source and electronic postprocessing of the images. We achieve 100-nm optical resolution, an improvement by a factor of more than 2 compared with standard fluorescence microscopy and of 1.5 compared with confocal scanning. PMID- 10840058 TI - Transposon stability and a role for conjugational transfer in adaptive mutability. AB - Lac(+) revertants of Escherichia coli that occur after prolonged nonlethal selection display a high frequency of transposon loss when the transposon Tn10 and the reverting lacI33 allele are linked on an F'128 episome. As many as 20% of the Lac(+) revertants are sensitive to tetracycline, about half because of transposon loss, nearly all by precise excision, and the remainder because of amplification of both the transposon and the linked lac allele. Lethality of the amplified products in the presence of tetracycline is a peculiarity of the tetA gene at high gene dosage. The selective conditions on lactose medium result in 10% transposon-free revertants, whether or not a requirement for conjugal DNA transfer is imposed. In addition, a similar fraction, about 5% of Lac(-) unreverted colonies that are products of transfer between cells experiencing nonlethal selection are also tetracycline-sensitive, and all are attributable to loss of the Tn10 transposon. These results suggest the possibility that the high frequency of transposon loss is a consequence of conjugal transfer, making this loss a marker for that transfer. We suggest that conjugal DNA transfer may be a prominent feature in the mutability process that occurs during nonlethal selection and that the subset of bacteria displaying hypermutability are those that experience such transfer. PMID- 10840059 TI - DNA delivery by phage as a strategy for encapsulating toroidal condensates of arbitrary size into liposomes. AB - We report a strategy for encapsulating and condensing DNA. When T5 phage binds to its membrane protein receptor, FhuA, its double stranded DNA (120,000 bp) is progressively released base pair after base pair in the surrounding medium. Using cryoelectron microscopy, we have visualized the structures formed after T5 phage DNA is released into neutral unilamellar proteoliposomes reconstituted with the receptor FhuA. In the presence of spermine, toroidal condensates of circumferentially wrapped DNA were formed. Most significantly, the sizes of these toroids were shown to vary, from 90 to 200 nm in their outer diameters, depending on the number of DNA stands transferred. We have also analyzed T5 DNA release in bulk solution containing the detergent-solubilized FhuA receptor. After DNA release in a spermine containing solution, huge DNA condensates with a diameter of about 300 nm were formed containing the DNAs from as many as 10-20 capsids. At alkaline pH, the condensates appeared as large hollow cylinders with a diameter of 200 nm and a height of 100-200 nm. Overall, the striking feature of our experiments is that, because of the progressive release of DNA from the phage capsid, the mechanism of toroid formation is fundamentally different from that in the classical studies in which highly dilute, "naked" DNA is condensed by direct addition of polyvalent cations; as a consequence, our method leads to toroids of arbitrary size. PMID- 10840060 TI - Noninvasive gene targeting to the brain. AB - Gene therapy of the brain is hindered by the presence of the blood-brain barrier (BBB), which prevents the brain uptake of bloodborne gene formulations. Exogenous genes have been expressed in the brain after invasive routes of administration, such as craniotomy or intracarotid arterial infusion of noxious agents causing BBB disruption. The present studies describe the expression of an exogenous gene in brain after noninvasive i.v. administration of a 6- to 7-kb expression plasmid encoding either luciferase or beta-galactosidase packaged in the interior of neutral pegylated immunoliposomes. The latter are conjugated with the OX26 mAb to the rat transferrin receptor, which enables targeting of the plasmid DNA to the brain via the endogenous BBB transferrin receptor. Unlike cationic liposomes, this neutral liposome formulation is stable in blood and does not result in selective entrapment in the lung. Luciferase gene expression in the brain peaks at 48 h after a single i.v. administration of 10 microg of plasmid DNA per adult rat, a dose that is 30- to 100-fold lower than that used for gene expression in rodents with cationic liposomes. beta-Galactosidase histochemistry demonstrated gene expression throughout the central nervous system, including neurons, choroid plexus epithelium, and the brain microvasculature. In conclusion, widespread gene expression in the brain can be achieved by using a formulation that does not employ viruses or cationic liposomes, but instead uses endogenous receptor mediated transport pathways at the BBB. PMID- 10840061 TI - Recombinant tobacco mosaic virus movement protein is an RNA-binding, alpha helical membrane protein. AB - The 30-kDa movement protein (MP) is essential for cell-cell spread of tobacco mosaic virus in planta. To explore the structural properties of MP, the full length recombinant MP gene was expressed in Escherichia coli, and one-step purification from solubilized inclusion bodies was accomplished by using anion exchange chromatography. Soluble MP was maintained at >4 mg/ml without aggregation and displayed approximately 70% alpha-helical conformation in the presence of urea and SDS. A trypsin-resistant core domain of the MP had tightly folded tertiary structure, whereas 18 aa at the C terminus of the monomer were rapidly removed by trypsin. Two hydrophobic regions within the core were highly resistant to proteolysis. Based on results of CD spectroscopy, trypsin treatment, and MS, we propose a topological model in which MP has two putative alpha-helical transmembrane domains and a protease-sensitive carboxyl terminus. PMID- 10840062 TI - Cloning of the Arabidopsis WIGGUM gene identifies a role for farnesylation in meristem development. AB - Control of cellular proliferation in plant meristems is important for maintaining the correct number and position of developing organs. One of the genes identified in the control of floral and apical meristem size and floral organ number in Arabidopsis thaliana is WIGGUM. In wiggum mutants, one of the most striking phenotypes is an increase in floral organ number, particularly in the sepals and petals, correlating with an increase in the width of young floral meristems. Additional phenotypes include reduced and delayed germination, delayed flowering, maturation, and senescence, decreased internode elongation, shortened roots, aberrant phyllotaxy of flowers, aberrant sepal development, floral buds that open precociously, and occasional apical meristem fasciation. As a first step in determining a molecular function for WIGGUM, we used positional cloning to identify the gene. DNA sequencing revealed that WIGGUM is identical to ERA1 (enhanced response to abscisic acid), a previously identified farnesyltransferase beta-subunit gene of Arabidopsis. This finding provides a link between protein modification by farnesylation and the control of meristem size. Using in situ hybridization, we examined the expression of ERA1 throughout development and found it to be nearly ubiquitous. This extensive expression domain is consistent with the pleiotropic nature of wiggum mutants and highlights a broad utility for farnesylation in plant growth and development. PMID- 10840063 TI - Replication-induced transcription of an autorepressed gene: the replication initiator gene of plasmid P1. AB - The replication origin of plasmid P1 contains an array of five repeats (iterons) that bind the plasmid-encoded initiator RepA. Within the array lies the repA promoter, which becomes largely repressed on RepA binding (autorepression). One might expect that extra iterons produced on plasmid replication would titrate RepA and release the repression. The promoter, however, is induced poorly by extra iterons. The P1 copy number is reduced by extra iterons in the presence of the autorepressed repA gene but not when additional RepA is provided from constitutive sources. It has been proposed that the iteron-bound RepA couples with the promoter-bound RepA and thereby maintains repression. Although not the product of replication, we find that the act of replication itself can renew RepA synthesis. Replication apparently cleans the promoter of bound RepA and provides a window of opportunity for repA transcription. We propose that replication induced transcription is required to ensure initiator availability in a system that is induced poorly when challenged with additional initiator binding sites. PMID- 10840064 TI - Human centromere protein A (CENP-A) can replace histone H3 in nucleosome reconstitution in vitro. AB - Centromere protein A (CENP-A) is a variant of histone H3 with more than 60% sequence identity at the C-terminal histone fold domain. CENP-A specifically locates to active centromeres of animal chromosomes and therefore is believed to be a component of the specialized centromeric nucleosomes on which the kinetochores are assembled. Here we report that CENP-A, highly purified from HeLa cells, can indeed replace histone H3 in a nucleosome reconstitution system mediated by nucleosome assembly protein-1 (NAP-1). The structure of the nucleosomes reconstituted with recombinant CENP-A, histones H2A, H2B, and H4, and closed circular DNAs had the following properties. By atomic force microscopy, "beads on a string" images were obtained that were similar to those obtained with nucleosomes reconstituted with four standard histones. DNA ladders with repeats of approximately 10 bp were produced by DNase I digestion, indicating that the DNA was wrapped round the protein complex. Mononucleosomes isolated by glycerol gradient sedimentation had a relative molecular mass of approximately 200 kDa and were composed of 120-150 bp of DNA and equimolar amounts of CENP-A, and histones H4, H2A, and H2B. Thus, we conclude that CENP-A forms an octameric complex with histones H4, H2A, and H2B in the presence of DNA. PMID- 10840065 TI - The RecD subunit of the Escherichia coli RecBCD enzyme inhibits RecA loading, homologous recombination, and DNA repair. AB - The RecBCD enzyme is required for homologous recombination and DNA repair in Escherichia coli. The structure and function of RecBCD enzyme is altered on its interaction with the recombination hotspot Chi (5'-GCTGGTGG-3'). It has been hypothesized that the RecD subunit plays a role in Chi-dependent regulation of enzyme activity [Thaler, D. S., Sampson, E., Siddiqi, I., Rosenberg, S. M., Stahl, F. W. & Stahl, M. (1988) in Mechanisms and Consequences of DNA Damage Processing, eds. Friedberg, E. & Hanawalt, P. (Liss, New York), pp. 413-422; Churchill, J. J., Anderson, D. G. & Kowalczykowski, S. C. (1999) Genes Dev. 13, 901-911]. We tested the hypothesis that the RecD subunit inhibits recombination by deleting recD from the nuclease- and recombination-deficient mutant recB(D1080A)CD. We report here that the resulting strain, recB(D1080A)C, was proficient for recombination and DNA repair. Recombination proficiency was accompanied by a change in enzyme activity: RecB(D1080A)C enzyme loaded RecA protein onto DNA during DNA unwinding whereas RecB(D1080A)CD enzyme did not. Together, these genetic and biochemical results demonstrate that RecA loading by RecBCD enzyme is required for recombination in E. coli cells and suggest that RecD interferes with the enzyme domain required for its loading. A nuclease dependent signal appears to be required for a change in RecD that allows RecA loading. Because RecA loading is not observed with wild-type RecBCD enzyme until it acts at a Chi site, our observations support the view that RecD inhibits recombination until the enzyme acts at Chi. PMID- 10840067 TI - Absence of toxicity of Bacillus thuringiensis pollen to black swallowtails under field conditions. AB - A single laboratory study on monarch butterflies has prompted widespread concern that corn pollen, engineered to express Bacillus thuringiensis (Bt) endotoxin, might travel beyond corn fields and cause mortality in nontarget lepidopterans. Among the lepidopterans at high potential risk from this technology is the black swallowtail butterfly, Papilio polyxenes, whose host plants in the midwestern U. S. are located principally in narrow strips between roads and crop fields. A field study was performed to assess whether mortality of early instar black swallowtails was associated either with proximity to a field of Bt corn or by levels of Bt pollen deposition on host plants. Potted host plants were infested with first instar black swallowtails and placed at intervals from the edge of a field of Bt corn (Pioneer 34R07 containing Monsanto event 810) at the beginning of anthesis. We confirmed by ELISA that pollen from these plants contained Cry1Ab endotoxin (2.125 +/- 0.289 ng/g). Although many of the larvae died during the 7 days that the experiments were run, there was no relationship between mortality and proximity to the field or pollen deposition on host plants. Moreover, pollen from these same plants failed to cause mortality in the laboratory at the highest pollen dose tested (10,000 grains/cm(2)), a level that far exceeded the highest pollen density observed in the field (200 grains/cm(2)). We conclude that Bt pollen of the variety tested is unlikely to affect wild populations of black swallowtails. Thus, our results suggest that at least some potential nontarget effects of the use of transgenic plants may be manageable. PMID- 10840068 TI - New strategies for the treatment of acute myelogenous leukemia: differentiation induction--present use and future possibilities. AB - A differentiation block and an accumulation of immature myeloid cells characterize acute myelogenous leukemia (AML). However, native AML cells usually show some morphological signs of differentiation that allow a classification into different subsets, and further differentiation may be induced by exposure to various soluble mediators, for example, all-trans retinoic acid (ATRA) and several cytokines. Combination therapy with ATRA and chemotherapy should now be regarded as the standard treatment of the acute promyelocytic leukemia (APL) variant of AML. Although several agents can also induce leukemic cell differentiation for other AML subgroups, in vitro studies as well as clinical data have demonstrated that these agents often have heterogeneous effects on the leukemic progenitors. This makes the clinical impact of differentiation induction therapy for individual patients difficult to predict. However, differentiation induction should be regarded as a promising therapeutic approach, especially as a part of immunotherapy or in combination with intensive chemotherapy to increase the susceptibility of AML blasts to drug-induced apoptosis. Although the morphology-based French-American-British classification was used to identify APL as an AML subset that required a special treatment, it seems unlikely that this classification alone can be used to identify new subsets of AML patients with special therapeutic requirements. Future studies on differentiation induction in AML should therefore focus on A) the identification of therapeutic agents with more predictable effects; B) the use of clinical and laboratory parameters to define new subsets of AML patients in which differentiation induction has a predictable and beneficial effect, and C) the characterization of how AML blast sensitivity to drug-induced apoptosis is altered by differentiation induction. PMID- 10840066 TI - Regulation of oxidative stress-induced calcium release by phosphatidylinositol 3 kinase and Bruton's tyrosine kinase in B cells. AB - Hydrogen peroxide stimulates a tyrosine kinase-dependent calcium release from intracellular stores, which is assumed to be achieved through the activation of phospholipase Cgamma2 (PLCgamma2) via a tyrosine phosphorylation mechanism in B cells. Here we show that H(2)O(2) induces both tyrosine phosphorylation on PLCgamma2 and the activation of phosphatidylinositol 3-kinase (PI3K) in B cells, and that the phosphatidylinositol 3-kinase inhibitor, Wortmannin, partially inhibited the H(2)O(2)-induced calcium release without affecting tyrosine phosphorylation on PLCgamma2. Overexpression of human Bruton's tyrosine kinase (Btk), which was activated by H(2)O(2), almost completely overcame the inhibition of calcium release by Wortmannin. The reversal of Wortmannin's inhibition by enhancing Btk concentration seemed unique to the H(2)O(2)-mediated effect, because Btk failed to overcome the inhibition of Wortmannin on B cell receptor triggered calcium mobilization. Immunoblot analysis revealed that Btk formed stable complexes with several tyrosine-phosphorylated proteins, including PLCgamma2, only in Btk-overexpressed cells on H(2)O(2) stimulation. Together, our data are consistent with the notion that PIP3 and/or a high concentration of Btk target the activated PLCgamma2 to its substrate site for maximal catalytic efficiency. PMID- 10840069 TI - The use of thymidylate synthase inhibitors in the treatment of advanced colorectal cancer: current status. AB - The combination of 5-fluorouracil (5-FU) and leucovorin has been the unofficial "standard" therapy for patients with colorectal cancer for over a decade. Recently, however, a number of new agents targeted against the enzyme thymidylate synthase have been synthesized and are in various stages of development. The currently available thymidylate synthase inhibitors are discussed. Enormous efforts have been made over the years to improve the efficacy of 5-FU, the most popular of these agents. Biochemical modulation by leucovorin has been the most successful so far. Continuous infusion schedules also appear to be advantageous over bolus administration. However, marked intra- and interpatient variability, combined with nonlinear elimination kinetics and erratic oral bioavailability are relative limitations to further development of 5-FU. New oral 5-FU prodrugs such as UFT, S-1, and Capecitabine may help to overcome some of these difficulties. Eniluracil, a potent inhibitor of the enzyme dihydropyrimidine dehydrogenase, may also help by overcoming potential 5-FU resistance mechanisms, in addition to increasing its bioavailability. Of the antifolate-based inhibitors, Tomudex is in the most advanced stage of development. Similar efficacy with 5-FU and a convenient schedule may suggest a role in future combination regimens. It is quite likely that even the most optimal thymidylate synthase inhibition will have limitations in terms of clinical efficacy. Novel combinations of 5-FU or its analogs with agents that have different mechanisms of action (e.g., oxaliplatin, irinotecan) could provide important new opportunities for improving the outlook of patients with colorectal cancer. PMID- 10840070 TI - Maturation and lineage-specific expression of the coxsackie and adenovirus receptor in hematopoietic cells. AB - Adenovirus vectors have been used to transfer genes into both hematopoietic progenitor cells and tumor cells, including carcinoma cells that have metastasized to bone marrow (BM). However, the relative susceptibility of different subsets of hematopoietic cells is unknown. In permissive cells adenoviral-mediated gene transfer is mediated by the coxsackievirus and adenovirus receptor (CAR) protein and alpha(v) integrins expressed on the cell surface of the target cells. This prompted us to investigate the expression of CAR on subpopulations of hematopoietic cells, determine whether this protein played a role in adenovirus-mediated gene transfer of hematopoietic cells and whether we could modulate CAR to enhance gene transfer efficiency. In this report we show that CAR is expressed on approximately 40% of all human BM cells, including erythroid and myeloid cells, but not lymphoid cells. Of the CD34(+) cells, 10%-15% expressed CAR, but this did not include most colony-forming progenitor cells, nor the most primitive CD38(-) subpopulation. The presence of CAR correlated well with gene transfer efficiency, but we were unable to induce CAR expression on immature, noncommitted progenitor cells. In conclusion, our results show that primitive hematopoietic progenitor cells lack CAR expression, but that expression is acquired during erythroid and myeloid differentiation. PMID- 10840071 TI - Murine stromal cell line HESS-5 maintains reconstituting ability of Ex vivo generated hematopoietic stem cells from human bone marrow and cytokine-mobilized peripheral blood. AB - Human bone marrow (BM) or mobilized peripheral blood (mPB) CD34(+) cells have been shown to loose their stem cell quality during culture period more easily than those from cord blood (CB). We previously reported that human umbilical CB stem cells could effectively be expanded in the presence of human recombinant cytokines and a newly established murine bone marrow stromal cell line HESS-5. In this study we assessed the efficacy of this xenogeneic coculture system using human BM and mPB CD34(+) cells as materials. We measured the generation of CD34(+)CD38(-) cells and colony-forming units, and assessed severe-combined immunodeficient mouse-repopulating cell (SRC) activity using cells five days after serum-free cytokine-containing culture in the presence or the absence of a direct contact with HESS-5 cells. As compared with the stroma-free culture, the xenogeneic coculture was significantly superior on expansion of CD34(+)CD38(-) cells and colony-forming cells and on maintenance of SRC activity. The PKH26 study demonstrated that cell division was promoted faster in cells cocultured with HESS-5 cells than in cells cultured without HESS-5 cells. These results indicate that HESS-5 supports rapid generation of primitive progenitor cells (PPC) and maintains reconstituting ability of newly generated stem cells during ex vivo culture irrespective of the source of samples. This xenogeneic coculture system will be useful for ex vivo manipulation such as gene transduction to promote cell division and the generation of PPC and to prevent loss of stem cell quality. PMID- 10840072 TI - Low numbers of megakaryocyte progenitors in grafts of cord blood cells may result in delayed platelet recovery after cord blood cell transplant. AB - Delayed platelet recovery is an inherent problem with cord blood cell transplantation (CBCT). To investigate this problem, the number of human megakaryocyte (MK) progenitor cells in cord blood (CB; n = 24) was measured and compared with that in G-CSF-mobilized peripheral blood stem cells (PBSC; n = 25). The median numbers of colony-forming units for MK (CFU-MK) that were detected by a serum-free assay system in CB and peripheral blood (PB) were 26 (range, 6 102)/10(5) nucleated cells (NC) and 37 (2-540)/10(5) mononuclear cells (MNC), respectively. The numbers of colony-forming units for granulocyte/macrophage (CFU GM) were 88 (33-241)/10(5) NC in CB and 138 (6.3-1,250)/10(5) MNC in PB. The frequencies of CD34(+) cells in CB and PB were, respectively, 0.44% (0.10-1.07) and 0.98% (0.05-20.8). The numbers of CFU-MK in CB and PBSC were correlated with those of CD34(+) cells. The estimated number of infused CFU-MK in CBCT was 1/15 that of PBSC transplantation (PBSCT), based upon the above data and the widely used standard doses for both types of transplants. Further, the numbers of infused CFU-MK in patients who received allogeneic PBSCT at our institute were inversely correlated with the speed of platelet recovery. These data indicate that delayed platelet recovery after CBCT is simply due to the low number of CFU MK contained in grafts. PMID- 10840073 TI - In vitro proliferation potential of AC133 positive cells in peripheral blood. AB - AC133 antigen is a novel marker for human hematopoietic stem/progenitor cells. In this study, we examined the expression and proliferation potential of AC133(+) cells obtained from steady-state peripheral blood (PB). The proportion of AC133(+) cells in the CD34(+) subpopulation of steady-state PB was significantly lower than that of cord blood (CB), although that of cytokine-mobilized PB was higher than that of CB. The proliferation potential of AC133(+)CD34(+) and AC133( )CD34(+) cells was examined by colony-forming analysis and analysis of long-term culture-initiating cells (LTC-IC). Although the total number of colony-forming cells was essentially the same in the AC133(+)CD34(+) fraction as in the AC133( )CD34(+) fraction, the proportion of LTC-IC was much higher in the AC133(+)CD34(+) fraction. Virtually no LTC-IC were detected in the AC133( )CD34(+) fraction. In addition, the features of the colonies grown from these two fractions were quite different. Approximately 70% of the colonies derived from the AC133(+)CD34(+) fraction were granulocyte-macrophage colonies, whereas more than 90% of the colonies derived from the AC133(-)CD34(+) fraction were erythroid colonies. Furthermore, an ex vivo expansion study observed expansion of colony forming cells only in the AC133(+)CD34(+) population, and not in the AC133( )CD34(+) population. These findings suggest that to isolate primitive hematopoietic cells from steady-state PB, selection by AC133 expression is better than selection by CD34 expression. PMID- 10840074 TI - Hematopoietic repopulating ability of cord blood CD34(+) cells in NOD/Shi-scid mice. AB - Although umbilical cord blood (CB) is increasingly being used as an alternative to bone marrow (BM) as a source of transplantable hematopoietic stem cells (HSC), information on the hematopoietic repopulating ability of CB HSC is still limited. We recently established a xenotransplantation system in NOD/Shi-scid mice to evaluate human stem cell activity. In the present study, we transplanted 5 to 10 x 10(4) CB CD34(+) cells into six NOD/Shi-scid mice treated with anti-asialo GM1 antiserum to investigate the hematopoietic repopulating ability of CB. The BM of all recipients contained human CD45(+) cells 10 to 12 weeks after the transplantation (43.8 +/- 17.7%). Clonal culture of the recipient BM cells revealed the formation of various types of human hematopoietic colonies, including myelocytic, erythroid, megakaryocytic, and multilineage colonies, indicating that CB HSC can differentiate into hematopoietic progenitors of various lineages. However, the extent of the differentiation and maturation differed with each lineage. CD13(+)/CD14(+)/CD33(+) myelocytic cells were mainly repopulated in BM and peripheral blood (PB). While CD41(+) megakaryocytic cells and platelets were present, few glycophorin A(+)CD71(+) or hemoglobin alpha containing erythroid cells were detected. CD19(+) B cells were the most abundantly repopulated in NOD/Shi-scid mice, but their maturational stage differed among the hematopoietic organs. Most of the BM CD19(+) cells were immature B cells expressing CD10 but not surface immunoglobulin (Ig) M, whereas more mature CD19(+)CD10(-) surface IgM(+) B cells were predominantly present in spleen and PB. CD3(+) T cells were not detected even in the recipient thymus. The transplantation to the NOD/Shi-scid mouse may provide a useful tool for evaluating the repopulating ability of transplantable human HSC. PMID- 10840075 TI - Mechanisms mediating the inhibitory effect of all-trans retinoic acid on primitive hematopoietic stem cells in human long-term bone marrow culture. AB - All-trans retinoic acid (RA) has generally been found to stimulate late committed (colony-forming unit- granulocyte, macrophage [CFU-GM]) and inhibit early (CFU Blast) normal human myeloid progenitor cells. The present study provides the first evidence that the pharmacological concentration of 1 microM RA, exerts an inhibitory effect on the proliferation of functional human primitive hemopoietic stem cells (cobblestone area-forming cell [CFAC]) in long-term bone marrow cultures. Treatment of four-week confluent bone marrow culture with 1 microM RA for five days significantly reduced week 4 CAFC from 88 +/- 10 in control cultures to only 52 +/- 12 per 10(5) cells, p < 0.01. Quantitative enzyme-linked immunosorbent assay measurement of interleukin 6 (IL-6) and IL-11 produced from the four-week bone marrow stroma culture revealed only a slight and moderate increase of IL-6 and IL-11 production after treatment with RA. On the other hand, treatment with RA profoundly increased the soluble receptor gp130 released from the four-week bone marrow stroma by 7.5-fold from only 145 +/- 2.1 pg per ml in control cultures to 1,069.9 +/- 3.8 pg per ml in RA-treated cultures. A similar marked increase in the soluble adhesion molecules ICAM-1, and to a lesser extent VCAM-1, released from the four-week bone marrow stroma was observed after RA treatment. IL-6 has been implicated in the inhibitory effect of RA in several human hemopoietic and nonhemopoietic cells. The common transducing signal chain gp130, for all receptors of the IL-6 cytokine family, is expressed in most primitive human hemopoietic CD34(+) cells and its signaling was shown to synergize with other hemopoietic cytokines to expand primitive human hemopoietic stem cells. Recently, soluble gp130 was shown to be a natural potent antagonist of the human IL-6 cytokine family by binding the ligand and thereby reducing its bioavailability. The profound and rapid 7.5-fold increase in the natural antagonist of human IL-6 cytokine family after RA treatment could abrogate the gp130 signaling required for proliferation and/or expansion of human primitive hemopoietic stem cells and lead to the observed inhibitory effect of RA on CAFC. Both adhesion molecules VCAM-1 and ICAM-1 mediate human hemopoietic stem cell adhesion to marrow stroma. The present significant increase in the soluble form of these adhesion molecules after RA treatment could exert a significant antagonist effect on their function and hence may impair CAFC adhesion to marrow stroma. In conclusion, the RA inhibitory effect on the proliferation of primitive human hemopoietic stem cells could be mediated through: A) an impaired hemopoietic stem cell adhesion due to the significant increase in soluble adhesion molecules released from the marrow stroma after RA treatment, and B) a significantly reduced gp130 signaling that is necessary for stem cell proliferation due to the natural antagonistic effect of the profoundly increased level of soluble gp130 released from the marrow stroma after treatment with RA. PMID- 10840076 TI - Cellular suicide therapy of malignant disease. AB - Adoptive cellular therapy is developing as a supplement or alternative to chemotherapy and/or radiation for malignant disease. Our focus is two ongoing clinical studies with transgeneic (genetically altered) cellular therapy; one uses allogeneic (from another person) lymphocytes to treat leukemia, and the second uses xenogeneic (from another species) fibroblast cells genetically altered to contain a toxin-producing suicide gene to treat ovarian cancer. Allogeneic donor lymphocyte infusions (DLI) are known to induce remission of hematologic malignancies. However, the toxicity associated with DLI is related to graft-versus-host-disease, which is due to donor lymphocytes attacking normal tissue in the recipient. Therefore, we have taken the approach of infusing DLI that have been modified to contain a latent suicide gene to treat leukemia. To treat ovarian cancer, we used xenogeneic nonimmune fibroblast-derived cells to deliver a tumor-directed cytotoxic gene to carcinoma cells. These cells release HStk transgene retroviruses that in turn transduce replicating tumor cells but not quiescent epithelium, rendering the tumor selectively susceptible to ganciclovir-mediated killing. These initial trials summarize the early stage of allogeneic/xenogeneic adoptive cellular therapy for cancer, and although the data are limited, it is encouraging to see some patients with evidence of antitumor responses. Advances in our understanding of the basic science of these treatments, together with improvements in the technology of vector design, will be required to streamline these methodologies into broader application. PMID- 10840077 TI - The molecular perspective: cyclooxygenase-2. PMID- 10840078 TI - Lowering PSA cutoffs to enhance detection of curable prostate cancer. PMID- 10840079 TI - A PSA threshold of 4.0 ng/mL for early detection of prostate cancer: the only rational approach for men 50 years old and older. PMID- 10840080 TI - Fibroblast growth factor 2: its structure and property, paracrine function, tumor angiogenesis, and prostate-related mitogenic and oncogenic functions. PMID- 10840081 TI - Laparoscopic donor nephrectomy: current role in renal allograft procurement. PMID- 10840082 TI - Sildenafil effects on sexual and cardiovascular responses in women with spinal cord injury. AB - OBJECTIVES: Sexual dysfunction is common in women with spinal cord injuries (SCIs) and other neurologic conditions. Sildenafil has previously been shown to be safe and effective in the treatment of erectile dysfunction due to SCI. This study is the first to evaluate the sexual and cardiovascular effects of sildenafil in women with SCIs in a controlled, laboratory setting. METHODS: Nineteen premenopausal women with SCIs were randomly assigned to receive either sildenafil (50 mg) or placebo in a double-blind, crossover design study. Physiologic and subjective measures of sexual response, heart rate, and blood pressure were recorded during baseline and sexual stimulation conditions. Adverse events were also recorded. RESULTS: Significant increases in subjective arousal (SA) were observed with both drug (P <0.01) and sexual stimulation conditions (P <0.001), and a borderline significant (P <0.07) effect of drug administration on vaginal pulse amplitude (VPA) was noted. Maximal responses occurred when sildenafil was combined with visual and manual sexual stimulation. Cardiovascular data showed modest increases in heart rate (+/-5 bpm) and mild decreases in blood pressure (+/-4 mm Hg) across all stimulation conditions, consistent with the peripheral vasodilatory mechanism of the drug. Sildenafil was well tolerated with no evidence of significant adverse events. CONCLUSIONS: Findings suggest that sildenafil may partially reverse the sexual dysfunction commonly associated with SCI in women. Consistent with previous findings in men, the sexual effects of the drug were most evident under conditions of optimal stimulation. Mild, clinically insignificant cardiovascular effects were also noted. Further large-scale studies of sildenafil's effects in women with neurogenic sexual dysfunction are strongly indicated. PMID- 10840083 TI - Determination of stone composition by noncontrast spiral computed tomography in the clinical setting. AB - OBJECTIVES: Several investigators have evaluated noncontrast computed tomography (NCCT) in predicting stone composition in vitro. We assessed NCCT in predicting stone composition in patients presenting to our emergency room with flank pain and stone disease. METHODS: One hundred twenty-nine patients presenting to our university hospital with flank pain underwent renal colic protocol NCCT scans at the request of the emergency room physicians. A General Electric, high-speed advantage CT scanner was used at 120 kV, 200 mA, and 1.4:1 pitch, with collimation varying between 3 and 5 mm. Ninety-nine patients with predominantly (greater than 50%) calcium oxalate or uric acid composition after either stone passage or stone removal were identified. Each scan was analyzed by one of two radiologists, who determined the predominant attenuation for each stone. Stones once passed or retrieved were analyzed by Urocor Laboratories. The attenuation and attenuation/size ratio (peak attenuation/size in millimeters) were compared with the results of the stone analysis. RESULTS: Eighty-two calculi predominantly composed of calcium oxalate and 17 calculi predominantly composed of uric acid were identified in 99 patients. The calculi ranged in size from 1 to 28 mm. A significant difference (P = 0.017, unpaired t test) was found between the Hounsfield measurement of uric acid calculi (mean 344 +/- 152 HU) and the Hounsfield measurement of calcium oxalate calculi (mean 652 +/- 490 HU). If only the Hounsfield units from stones 4 mm or larger were compared, the data were even more compelling (P = 0.002). However, using an attenuation/size ratio cutoff of greater than 80, the negative predictive value was 99% that a stone would be predominantly calcium oxalate. CONCLUSIONS: Using peak attenuation measurements and the attenuation/size ratio of urinary calculi from NCCT, we were able to differentiate between uric acid and calcium oxalate stones. PMID- 10840084 TI - Seminal vesicle-sparing radical prostatectomy: a novel concept to restore early urinary continence. AB - OBJECTIVES: Urinary incontinence after radical prostatectomy continues to be a distressing problem, even with preservation of the neurovascular bundles and meticulous apical dissection. Recent studies suggest that motor and sensory components of the pelvic nerve may be affected by surgery, since both components are anatomically located in intimate contact with the seminal vesicles. We propose seminal vesicle-sparing radical prostatectomy to preserve pelvic innervation and improve the rate of urinary continence. METHODS: Fifty-four patients were enrolled in this prospective study. A standard retropubic radical prostatectomy was performed in 34 patients. A seminal vesicle-sparing radical prostatectomy was performed in a pilot series of 20 consecutive patients. The seminal vesicle tip and surrounding tissue were preserved and carefully handled. In all patients, a modified pad test and posterior urethral sensory threshold test were performed preoperatively and 6 weeks and 6 months postoperatively and correlated with urinary continence. RESULTS: The intraoperative preservation of the seminal vesicle tip was possible in all patients in this pilot series (n = 20). In the seminal vesicle-sparing radical prostatectomy group, the continence rate was 60% after 6 weeks and 95% after 6 months. These rates were significantly higher than the continence rates in the standard prostatectomy group (18% and 82% at 6 weeks and 6 months, respectively). The sensory threshold levels in the seminal vesicle-sparing group were similar to the preoperative values and were significantly lower than the postoperative threshold levels in the standard prostatectomy group. CONCLUSIONS: Seminal vesicle tip-sparing radical prostatectomy may be a surgical option to preserve pelvic innervation and maintain urinary continence after radical prostatectomy. Further randomized studies are necessary to elucidate the impact of seminal vesicle-sparing radical prostatectomy on restoration of urinary continence. PMID- 10840085 TI - Impact of body weight on urinary electrolytes in urinary stone formers. AB - OBJECTIVES: Obesity increases the risk of developing chronic medical conditions such as diabetes mellitus, hypertension, and coronary artery disease. We performed a retrospective review of a large data base on urinary stones to determine if differences are found in urine and serum chemistries among obese and nonobese stone-forming patients. The effect of body weight on stone recurrence among urinary stone formers was also determined. METHODS: A national data base containing serum biochemical profiles, 24-hour urine specimens, and standardized questionnaires was retrospectively evaluated from 5942 consecutive patients with urinary stone disease. Stone-forming patients were classified by body weight: nonobese men, less than 100 kg and nonobese women, less than 85 kg; intermediate men, 100 to 120 kg and intermediate women, 85 to 100 kg; and obese men, more than 120 kg and obese women, more than 100 kg. RESULTS: Obese stone formers comprised 6.8% (n = 404) of the patient population. The mean weight in the nonobese and obese groups was 81 kg versus 134 kg, respectively, for men and 64 kg versus 112 kg, respectively, for women. Obese patients represented 3.8% of the male and 12.6% of the female population. Obese patients had increased urinary excretion of sodium, calcium, magnesium, citrate, sulfate, phosphate, oxalate, uric acid, and cystine; obesity was associated with increased urinary volumes and urine osmolality compared with the nonobese patients. Obese men had increased concentration of urinary sodium, oxalate, uric acid, sulfate, and phosphate when corrected for urinary volume. Obese women had increased concentrations of sodium, uric acid, sulfate, phosphate, and cystine. The mean number of stone episodes in nonobese versus obese men was similar (3.55 and 3.56), whereas mean stone episodes were 2.93 and 3.38 (P = 0.045) for nonobese versus obese women. CONCLUSIONS: Among known stone formers, obesity is associated with unique changes in both serum and urinary chemistries. These changes are associated with an increased incidence of urinary stone episodes in obese women but not in obese men. PMID- 10840086 TI - Survey of neuromuscular injuries to the patient and surgeon during urologic laparoscopic surgery. AB - OBJECTIVES: Laparoscopy may be complicated by neuromuscular injuries, both to the patient and to the surgeon. We used a survey to estimate the incidence of these injuries during urologic laparoscopic surgery, to assess risk factors for these injuries, and to determine preventive measures. METHODS: A survey of neuromuscular injuries associated with laparoscopy submitted to 18 institutions in the United States was completed by 18 attending urologists from 15 institutions. RESULTS: From among a total of 1651 procedures, there were 46 neuromuscular injuries in 45 patients (2.7%), including abdominal wall neuralgia (14), extremity sensory deficit (12), extremity motor deficit (8), clinical rhabdomyolysis (6), shoulder contusion (4), and back spasm (2). Neuromuscular injuries were twice as common with upper retroperitoneal as with pelvic laparoscopy (3. 1% versus 1.5%). Among patients with neuromuscular injuries, those with rhabdomyolysis were heavier (means 91 versus 80 kg) and underwent longer procedures (means 379 versus 300 minutes), and those with motor deficits were older (means 51 versus 42 years of age). Of the surgeons, 28% and 17% reported frequent neck and shoulder pain, respectively. CONCLUSIONS: Although not common, neuromuscular injuries during laparoscopy do contribute to morbidity. Abdominal wall neuralgias, injuries to peripheral nerves, and joint or back injuries likely occur no more frequently than during open surgery, but risk of rhabdomyolysis may be increased. Positioning in a partial rather than full flank position may reduce the incidence of some injuries. Measures to reduce neuromuscular strain on the surgeon during laparoscopy should be considered. PMID- 10840087 TI - Delayed repair of post-traumatic posterior urethral distraction injuries: long term results. AB - OBJECTIVES: There is still controversy regarding the treatment of post-traumatic posterior urethral distraction injuries. Initial suprapubic cystostomy and delayed perineal urethral reconstruction has been considered the reference standard. In this report, we review our experience with delayed perineal urethral reconstruction, with a focus on the long-term outcome and complications. METHODS: A total of 77 men with posterior urethral distraction injury due to pelvic trauma underwent reconstruction with delayed perineal approach. In all cases, the area of fibrosis was aggressively excised, the corpus spongiosum was mobilized, and a tension-free, spatulated end-to-end anastomosis was achieved by splitting the corporeal bodies in 66.2% and by an additional perineally performed inferior pubectomy in 49.3% of the patients. The median time from injury to surgical repair was 12 months. The preoperative evaluation consisted of combined antegrade and retrograde cystourethrograms and cystourethrography. A detailed sexual history was obtained in 58 patients (75.3%). RESULTS: After a mean follow-up of 47 months (range 15 months to 14 years), the urethral continuity was adequate in 94. 8%; however, 2 patients required a perineal surgical revision (total of 79 operations). Postoperative incontinence was observed in 7 (9. 1%) of 77 patients. Postoperative erectile dysfunction was noted in 16.2% of patients who were known to be potent by history before surgery. CONCLUSIONS: Our results support the belief that delayed perineal reconstruction with extensive excision of fibrosis and a tension-free, spatulated end-to-end anastomosis is a successful treatment alternative for posterior urethral distraction defects, with acceptable morbidity. PMID- 10840088 TI - Evaluation of male sexual function after pelvic trauma by the International Index of Erectile Function. AB - OBJECTIVES: To assess the multifaceted male sexual function after pelvic ring fracture. METHODS: A cross-sectional study of male sexual function after pelvic ring fractures was conducted, using the International Index of Erectile Function (IIEF). The department of traumatology database was scanned (June 1996 to April 1999) for 30 to 70-year-old male patients with pelvic fracture. Seventy-six consecutive, literate patients were then contacted by mail. IIEF domain scores were calculated for all responders. Cappelleri's method for identification and grading of erectile dysfunction was applied for patients sexually active within the past 4 weeks. Student's t test was used to compare the domain scorings of patients with those of the control population used for the IIEF psychometric validation. Relationships between IIEF results and patient characteristics were sought by Spearman's rank correlation coefficient for quantitative variables and Student's t test for classes. RESULTS: Forty-six patients answered (60.1% response rate). Thirty-seven patients had experienced sexual intercourse in the past 4 weeks during which 11 patients (29.7%) had exhibited various degrees of impaired erection. As a whole, compared with the published controls a significant decrease in overall satisfaction (P <0.05) was demonstrated. Pubic diastasis was further related to impaired erectile function and overall satisfaction; we suggest that cavernosal nerves might be damaged at the time of diastasis. CONCLUSIONS: This study evidenced the impairment of sexual overall satisfaction after pelvic trauma and the specific decrease in erectile function and erection firmness and confidence associated with pubic diastasis. The IIEF questionnaire might be considered at the time of rehabilitation to identify those patients that could benefit from supportive treatments. PMID- 10840089 TI - Thoracoabdominal radical nephrectomy: is a postoperative thoracostomy tube necessary? AB - OBJECTIVES: To report our results of patients undergoing thoracoabdominal radical nephrectomy without intraoperative placement of a thoracostomy tube. It has been routine in our hospital to not place a thoracostomy tube in patients undergoing thoracoabdominal radical nephrectomy since 1988. METHODS: We conducted a retrospective review of 47 thoracoabdominal radical nephrectomies performed from January 1988 through November 1998 at our institution. Of the 47 patients, 39 did not have a thoracostomy tube placed intraoperatively; the other 8 patients did. The development of all postoperative complications, length of hospital stay, and hospital charges were noted. RESULTS: No postoperative mortality was noted in our study. Of the 47 patients in the study, 20 patients had a total of 29 complications. The overall number of complications was not increased in the group without a thoracostomy tube compared with the group with a thoracostomy tube (P = 0.104). No patient treated without a thoracostomy tube required subsequent placement of a tube for persistent pneumothorax. The mean length of hospital stay in patients with a thoracostomy tube after radical nephrectomy was 9.14 +/- 2.65 days; in patients without a thoracostomy tube, the mean length of stay was 7.07 +/- 3.97 days (P = 0.071). CONCLUSIONS: In patients without parietal pleural injury, thoracoabdominal radical nephrectomy without the placement of a thoracostomy tube can be performed safely and effectively, with a low risk of postoperative complications and a decrease in the overall hospital stay and hospital charges. PMID- 10840090 TI - Continent urinary reconstruction versus ileal conduit: a contemporary single institution comparison of perioperative morbidity and mortality. AB - OBJECTIVES: To compare postoperative morbidity and mortality in a concurrent and contemporary series of patients who underwent radical cystectomy with ileal conduit versus orthotopic neobladder. METHODS: The data of 198 patients were reviewed, 117 with orthotopic reconstruction and 81 with ileal conduit during a 5 year time frame. Thirty-day morbidity, mortality, reoperative rates, and parameters associated with the surgical procedures were obtained from chart review. RESULTS: No perioperative or postoperative deaths occurred in either group. The median operative time for the ileal conduit was 201 minutes (range 140 to 373), and for the orthotopic neobladder, it was 270 minutes (range 230 to 425). The median blood loss was 389 and 474 mL, respectively. The median length of hospitalization was 8 days for the ileal conduit group and 7 days for the orthotopic neobladder group. Diversion-related complications recognized within 30 days that ultimately required a return to the operating room occurred in 3.4% of those with a neobladder and 1.2% of those with an ileal conduit. CONCLUSIONS: The orthotopic neobladder is a longer and technically more complex procedure than the ileal conduit procedure. However, no demonstrable difference in morbidity or perioperative complications were found between the two procedures in our review. PMID- 10840092 TI - Editorial comment PMID- 10840091 TI - Collagen injection therapy in elderly women: long-term results and patient satisfaction. AB - OBJECTIVES: To evaluate long-term results and patient satisfaction using collagen injection therapy in elderly women. METHODS: Periurethral injection of collagen using local anesthesia was performed on 58 women 65 years old or older (range 65 to 86, mean 73. 2) to treat stress urinary incontinence. All patients underwent urodynamic evaluation. Forty-nine patients (84.5%) had intrinsic sphincteric deficiency; 9 patients (15.5%) had genuine stress urinary incontinence. Twenty one patients (36.2%) had no urethral hypermobility using Q-tip testing, and 37 (63.8%) had urethral hypermobility. RESULTS: At 2 months after injection, the initial response was assessed: 28 patients (48.3%) were totally dry and 18 (31.0%) were socially continent. Therapy was unsuccessful in 12 (20. 7%). To achieve continence, 1 to 4 injections (mean 1.9) were required. The average total volume to achieve success was 14.6 mL. No significant differences were observed in outcome, volume injected, or number of injections in patients with versus without urethral hypermobility. At a mean follow-up of 24.4 months (range 8 to 43), of the 46 patients who achieved continence, 19 (41.3%) developed recurrent leakage and required reinjection. The average interval to recurrence was 7.9 months (range 2 to 16). Of the 19 patients reinjected, only 8 (42.1%) regained continence. The long-term success rate after repeated injections was 35 (60.3%) of 58. An independent examiner contacted 40 patients for telephone interview. To date, 25 of the patients contacted noted a moderate or maximal level of symptom improvement, and 18 reported continued improvement in quality of life. Thirty-six patients noted minimal difficulty with the procedure, and 34 would recommend the treatment. CONCLUSIONS: Collagen is a safe, moderately effective alternative to manage stress urinary incontinence in elderly women. Elderly patients should be counseled that approximately 40% will experience recurrent leakage, which may not resolve with reinjection. PMID- 10840093 TI - Reply by the authors PMID- 10840094 TI - Health-related quality of life with orthotopic neobladder versus ileal conduit according to the SF-36 survey. AB - OBJECTIVES: To compare health-related quality of life (HRQOL) in patients with a neobladder and in patients with an ileal conduit. METHODS: HRQOL was assessed using the SF-36 survey, supplemented with a questionnaire concerning micturition status. RESULTS: Patients (n = 36) with a neobladder were significantly younger at time of surgery and time of survey than patients with an ileal conduit (n = 20). Mean (+/- SD) follow-up periods for patients with a neobladder and with an ileal conduit were 31.3 +/- 33.1 and 44.8 +/- 30.7 months, respectively. No significant difference was apparent in any scale score between neobladder and ileal conduit groups. Role-physical functioning (RP) and role-emotional functioning (RE) scale scores in both neobladder and ileal conduit patients appeared to be below the general U.S. population norm. Patients with neobladder 65 years old or older (n = 17) showed significantly lower RP and RE scores than those younger than 65 years (n = 19; P <0.05). Duration of follow-up was not related to scale scores. Continence status did not measurably affect HRQOL. CONCLUSIONS: All scales concerning HRQOL except RP and RE were favorable with both neobladder and ileal conduit, and no significant differences were observed between these two types of urinary tract reconstruction. Generally, patients with a neobladder or an ileal conduit were satisfied with their overall quality of life and health. PMID- 10840095 TI - Comparison of biomechanical properties of periosteal suture fixation and bone anchor fixation to the pubic bone. AB - OBJECTIVES: To compare the relative strength of fixation using bone anchors (BAs) compared with direct suture placement into the periosteum. METHODS: The anterior bony pelvis was harvested from 21 female cadavers. In each pelvis, BA suture fixation was performed using Cinch anchors on one side of the pubic bone and direct periosteal suture fixation (PSF) on the contralateral side of the same pelvis. We used No. 1 polyproprolene suture for all cases. Using a hydraulic mechanical testing machine, all specimens were loaded in uniaxial tension until failure. RESULTS: Failure modes for BA-fixed pelves were as follows: 11 BA pull out, 1 midsuture failure, and 9 suture cut by BA. Failure modes for the PSF pelves were as follows: 6 suture pull-outs through the bone, 14 midsuture failures, and 1 suture cut at the bone. PSF pelves required significantly higher loads to induce failure compared with BA pelves (PSF 92.63 +/- 22.62 N, BA 71.32 +/- 19.76 N, P <0.0002). In many cases, both PSF and BA were adequate points of fixation, and the major mechanism of failure was suture rupture. In pelves with suture failure, the load to induce failure was significantly higher in the PSF group (PSF 105.06 +/- 12.55 N, BA 86.06 +/- 7.78 N, P <0.0025). When the suture failed, PSF was better because BA fixation actually broke some sutures. The load required to induce failure was higher in the PSF groups in 19 (90.5%) of 21 pelves. CONCLUSIONS: Biomechanical testing using permanent monofilament suture did not demonstrate a superiority of BA suture fixation to PSF fixation. PSF appears superior, since BAs induced suture failure in many cases. PMID- 10840096 TI - Editorial comment PMID- 10840097 TI - Reply by the authors PMID- 10840098 TI - Comparative evaluation of the BTAstat test, NMP22, and voided urine cytology in the detection of primary and recurrent bladder tumors. AB - OBJECTIVES: This prospective study was undertaken to evaluate the diagnostic efficacy of the BTAstat test and nuclear matrix protein (NMP22) compared with voided urine cytology (VUC) in the detection of primary and recurrent bladder cancer. METHODS: A total of 147 patients provided a single voided urine sample for the BTAstat test, NMP22, and cytology prior to cystoscopy. Eighty-five of them had no bladder cancer history, whereas the remaining 62 were monitored for superficial bladder cancer. A group of 21 healthy age-matched volunteers were also enrolled in the study. RESULTS: Bladder cancer was confirmed histologically in 99 patients, of which 62 had primary tumors and 37 had recurrent ones. The overall sensitivity and specificity were 71.7% and 56.5% for the BTAstat test, 62.6% and 73. 9% for NMP22, and 38.4% and 94.2% for VUC. The optimal threshold value for NMP22 calculated with receiver operating characteristics curve, was 8 U/mL. BTAstat test was significantly more sensitive than VUC in detecting bladder cancer in all stage and grade subgroups, except GIII. On the contrary, NMP22 was significantly more sensitive than VUC only in stage Ta, grade I and II patients. BTAstat test had higher but not significantly different sensitivity than NMP22. CONCLUSIONS: Our data indicate a superiority of both BTAstat test and NMP22 over VUC in the detection of bladder cancer. Comparing BTAstat test with NMP22, the former proved to be more sensitive, whereas the latter was more specific. Ruling out diseases with potential interference can increase the overall specificity of both tests. False-positive results of either test in patients followed up for bladder cancer seem to correspond to future recurrences. PMID- 10840099 TI - Gender differences in stage-adjusted bladder cancer survival. AB - OBJECTIVES: Gender differences have been observed in the prognosis of patients with bladder cancer. It has also been suggested that these differences are caused by a worse stage distribution at diagnosis among women. The purpose of this study was to evaluate whether women with bladder cancer have a worse prognosis even after adjustment for disease stage at first presentation. METHODS: Data on patients with bladder cancer diagnosed between 1973 and 1996 and registered by one of the nine population-based Surveillance, Epidemiology, and End Results (SEER) cancer registries in the United States (n = 80,305) were obtained from the National Cancer Institute public domain SEER*Stat 2.0 package. Similar data on patients with bladder cancer diagnosed between 1987 and 1994 and registered by two population-based registries in the Netherlands (n = 1722) were obtained through the Comprehensive Cancer Centers, Amsterdam and South. Survival rates adjusted for mortality owing to other causes (ie, relative survival) were calculated for men and women within each category of the American Joint Committee on Cancer (SEER data) and TNM (Netherlands data) stage groupings.Results. In the United States, the 5-year relative survival rate of male patients with bladder cancer was calculated to be 79.5% (95% confidence interval 79.0% to 80.0%). Among women, the 5-year relative survival rate was significantly worse: 73.1% (95% confidence interval 72.2% to 74.0%). The male versus female 5-year survival rate among stage groups I, II, III, and IV was 96.5% versus 93.7%, 65.5% versus 59.6%, 58.8% versus 49.6%, and 27.1% versus 15.2%, respectively. The (sparser) data from the Netherlands were less conclusive. Women with Stage II and Stage IV disease fared worse than men but the reverse seemed to be true in Stage I disease. CONCLUSIONS: Female patients with bladder cancer have a worse prognosis than male patients. It is unlikely that the difference can explained entirely by the more frequent diagnosis of higher stages at first presentation among women. PMID- 10840100 TI - Seminal oxidative stress in patients with chronic prostatitis. AB - OBJECTIVES: An association between prostatitis and male infertility has been suspected, yet is poorly understood. Prostatitis is often associated with granulocytes in the prostatic fluid that generate reactive oxygen species (ROS), known to impair male fertility. We compared ROS, the total antioxidant capacity (TAC), and a novel index of oxidative stress (ROS-TAC score) in patients with chronic prostatitis and in healthy controls. METHODS: Semen specimens from 36 men with chronic prostatitis (National Institutes of Health category IIIa), 8 men with prostatodynia (National Institutes of Health category IIIb), and 19 controls attending our urologic clinic were examined according to the World Health Organization criteria. Leukocytospermia was measured by the Endtz test (myeloperoxidase assay). ROS and TAC production was measured by chemiluminescence assay. A composite ROS-TAC score was also calculated in patients and controls. RESULTS: The sperm concentration, percentage of motility, and morphology among the groups did not differ. The mean +/- standard error log-transformed ROS level was significantly higher in patients with leukocytospermia (3.2 +/- 0.6) than in patients without leukocytospermia (1.8 +/- 0.2; P = 0.04) and controls (1.3 +/- 0.3, P = 0.01). TAC was significantly lower in patients with or without leukocytospermia (859.69 +/- 193.0 and 914.9 +/- 65.2, respectively) than in controls (1653.98 +/- 93.6, P = 0.001). The mean ROS-TAC score of controls (50.0 +/- 4.1) was significantly higher than those of patients with chronic prostatitis and leukocytospermia (8.2 +/- 9.2) and those without leukocytospermia (34.2 +/- 2.9; P <0.001). CONCLUSIONS: Men with chronic prostatitis or prostatodynia have seminal oxidative stress, irrespective of their leukocytospermia status. These observations may help shed light on the long-standing controversy surrounding prostatitis and infertility. PMID- 10840101 TI - Prospective randomized study of transurethral vaporization resection of the prostate using the thick loop and standard transurethral prostatectomy. AB - OBJECTIVES: Transurethral vaporization resection of the prostate (TUVRP) is a recent modification of the standard transurethral prostatectomy (TURP). The procedure uses one of the novel, thick resection loops coupled to augmented electrocutting energy. We evaluated the safety and efficacy of TUVRP in comparison with TURP. METHODS: Sixty-eight patients with prostatic outflow obstruction were prospectively randomized between equal TUVRP and TURP treatment groups. Safety parameters evaluated included changes in serum hemoglobin, hematocrit, and sodium 1 and 24 hours after resection. Operative time, catheterization time, and incidence of complications were noted. Efficacy parameters included evaluation by the International Prostate Symptom Score and maximum flow rate. RESULTS: Patients of both groups were balanced for the different baseline variables. One hour after TURP, patients had significantly lower levels of hemoglobin, hematocrit, and sodium (P = 0.03, 0.03, and 0. 01, respectively). The prostate resection weight was similar in both groups; however, the difference in the mean operative time was significant (TUVRP group 42.4 minutes and TURP group 35.9 minutes, P = 0.02). The postoperative catheterization time was significantly shorter for the TUVRP group (23.1 +/- 10.3 versus 36 +/- 17.3 hours, P <0.0001). All patients were followed up for an average of 9 months. The International Prostate Symptom Score was 4 +/- 3.4 and 5.6 +/- 3.1 and the maximum flow rate was 19 +/- 6.5 and 15.2 +/- 10 mL/s for the TUVRP and TURP groups, respectively; these differences were statistically significant (P = 0.03 and 0.01, respectively). Complications included urethral strictures (6 patients) and delayed hemorrhage with clot retention (2 patients); no differences in the incidence of complications were noted between the two groups. CONCLUSIONS: The results of the present study have demonstrated that TUVRP is as safe and efficacious as TURP in the treatment of men with prostatic outflow obstruction. The shorter catheterization time observed after TUVRP may be clinically significant, considering the demand for lower morbidity profiles by patients. The longer operative time in TUVRP was related to the slower motion of the Wing electrode needed to add the advantages of electrovaporization. PMID- 10840102 TI - Editorial comment PMID- 10840103 TI - Reply by the authors PMID- 10840104 TI - Correlation of histological inflammation in needle biopsy specimens with serum prostate- specific antigen levels in men with negative biopsy for prostate cancer. AB - OBJECTIVES: To reveal the possible contribution of histological inflammation within the prostate to the abnormal elevation of serum prostate-specific antigen (PSA) levels in patients with needle biopsy negative for prostate cancer. METHODS: We reviewed negative needle biopsy specimens obtained in 93 patients. The degree of acute and chronic inflammation as evaluated histologically was compared with serum PSA levels in conjunction with age and prostate volume. RESULTS: Both age (P <0.01) and prostate volume (P <0.0001) correlated significantly with serum PSA levels and were significantly greater in patients with abnormal serum PSA levels (greater than 4.0 ng/mL) than in those with normal serum PSA levels (4.0 ng/mL or less) (P <0.01). The presence of histological inflammation within the prostate also correlated significantly with serum PSA levels. Multiple regression analysis demonstrated prostate volume to be the only independent determinant of serum PSA levels (P <0.01). In patients with a prostate volume larger than 25 mL, only prostate volume correlated significantly with serum PSA levels (P <0. 05). On the other hand, the degree of acute inflammation as represented by polymorphonuclear leukocyte infiltration was the only parameter correlating significantly with serum PSA levels (P <0.05) in patients with a prostate volume smaller than 25 mL. CONCLUSIONS: Histologically defined acute inflammation within the prostate is a significant contributor to elevated serum PSA levels, especially in patients with small prostates. In the assessment of needle biopsy results negative for prostate cancer, it might be helpful to evaluate the degree of histological inflammation, especially in terms of the necessity of subsequent repeated biopsies. PMID- 10840105 TI - Impact of location and multifocality of positive surgical margins on disease-free survival following radical prostatectomy: a comparison between African-American and white men. AB - OBJECTIVES: Although the rate of positive surgical margins is higher in African American men (AAM) than in white men (WM), the impact of this difference on survival is not clear. Furthermore, it is unknown whether there are racial differences in the distribution of the positive surgical margins after radical retropubic prostatectomy (RRP). We investigated the differences between AAM and WM in terms of the site and multifocality of the positive surgical margins and their effect on disease-free survival (DFS) following RRP. METHODS: Between January 1991 and December 1995, 493 patients (288 WM and 205 AAM) were treated with RRP as monotherapy. Positive surgical margins were observed in 179 patients (86 WM and 93 AAM). Patients were divided in two groups: group 1 = WM and group 2 = AAM. The incidence and location of the positive surgical margins and their correlation with DFS were determined and compared. RESULTS: Overall, AAM had a higher rate of positive surgical margins than WM (48% versus 33%, respectively, P = 0.001). There was no significant difference in the frequency of multifocality of the positive margins (P = 0.4). Positive surgical margins were located significantly more often at the base in AAM (P = 0.015); however, the location of the positive surgical margins did not impact on DFS between groups. In those with multifocal positive surgical margins, AAM had a worse DFS compared with WM (P = 0.03). CONCLUSIONS: Race is an independent prognostic factor for DFS in patients with positive surgical margins. There were no differences in DFS between WM and AAM based on the margin location. In WM, prognostic factors for DFS in those with positive surgical margins were preoperative serum prostate-specific antigen, Gleason score, and pathologic stage. Conversely, in AAM none of these parameters were significant predictors of failure. PMID- 10840106 TI - Bladder neck-sparing modification of radical prostatectomy adversely affects surgical margins in pathologic T3a prostate cancer. AB - OBJECTIVES: To determine whether the bladder neck-sparing (BNS) modification of radical retropubic prostatectomy (RRP) alters the likelihood of positive surgical margins and postsurgical prostate cancer recurrence. METHODS: Surgical outcomes, as measured by pathologic margin status and progression-free survival, were evaluated in 751 consecutive RRP cases, among whom 222 underwent BNS technique. To reduce selection bias, comparison of positive margin rates between BNS and standard RRP was stratified by pathologic stage. Differences in surgical margin rates were assessed using the chi-square test, and effects of bladder neck preservation on prostate-specific antigen (PSA)-free survival were assessed, using multivariable Cox proportional hazards analysis. RESULTS: The clinical stage, Gleason score, and preoperative serum PSA profiles were similarly distributed between patients undergoing standard RRP and those undergoing the BNS modification. Surgical margins in the unstratified entire cohort were positive at rates similar to prior reports (28% BNS, 27% standard RRP). However, stratification by pathologic stage revealed that among pT3a cancers, BNS surgery was associated with significantly higher rates of positive surgical margins than was standard RRP (47% versus 20%; chi- square = 6.32, P = 0.01). Differences in positive margin rates were not seen between the two groups at other pathologic stages. The adverse effect of BNS technique on pT3a surgical margins was associated with a trend toward an adverse effect on PSA-free survival (Cox proportional hazards P = 0.016). CONCLUSIONS: The BNS modification of RRP can be associated with an increased rate of positive surgical margins specifically in cancers that have focally penetrated through the prostatic capsule (pT3a), with an associated trend toward decreased PSA-free survival in this group. BNS surgery may, therefore, compromise the ability to completely remove a subset of cancers focally penetrating the prostatic capsule. PMID- 10840107 TI - Probability of prostate cancer detection based on results of a multicenter study using the AxSYM free PSA and total PSA assays. AB - OBJECTIVES: The determination of the percentage of free prostate-specific antigen (%fPSA) enhances the specificity of prostate cancer (CaP) detection. This study was undertaken to assess the performance of %fPSA in differentiating benign prostate disease from CaP and to determine the CaP probability estimates using the AxSYM Free PSA and AxSYM Total PSA assays. METHODS: In this prospective study, 297 men, 50 years old or older, with a total PSA level between 4 and 10 ng/mL and a nonsuspicious digital rectal examination were enrolled at 10 clinical sites. All subjects underwent at least sextant prostate biopsies to establish the diagnosis. fPSA and total PSA (tPSA) levels were determined using the AxSYM Free PSA and AxSYM Total PSA assays. Percent fPSA values were compared with tPSA values to determine the appropriate cutoffs for prostate biopsy and to calculate the CaP probability estimates. RESULTS: The strongest predictor of CaP in a logistic regression model was %fPSA (odds ratio 2.29), which contributed significantly more than age or tPSA to the predictive model. In this study population, a %fPSA cutoff of 26.4% would have detected 96% of subjects with CaP (sensitivity) and would have eliminated 27.4% of unnecessary biopsies (specificity). CaP probability estimates ranged from 9% to 69% and increased as the %fPSA value decreased. Men with a %fPSA level of 10% or lower had a 69% probability of CaP, and men with a %fPSA level of greater than 26% had a 9% probability of CaP. CONCLUSIONS: Percent fPSA values can help differentiate CaP from benign prostate disease and reduce unnecessary biopsies in 27% of men 50 years old or older whose digital rectal examination was normal and whose tPSA level was between 4 and 10 ng/mL. A %fPSA result can assist the physician and patient in determining the probability of CaP and assessing the need for prostate biopsy. PMID- 10840108 TI - Anesthetic block of the dorsal penile nerve inhibits vibratory-induced ejaculation in men with spinal cord injuries. AB - OBJECTIVES: [corrected] We investigated which nerve pathways are necessary to achieve ejaculation using penile vibratory stimulation (PVS) in men with spinal cord injury (SCI). METHODS: Eight men with SCI were selected based on the presence of a bulbocavernosus reflex (BCR) and consistent antegrade ejaculation with PVS. Level of injury was cervical (4), upper thoracic (4), and lower thoracic (1). Mean age was 30.4 years (range 22 to 38). Usual responses to PVS included autonomic dysreflexia (4), erection (4), and consistent somatic responses such as abdominal contractions (8). Local anesthesia of the dorsal penile nerves (penile block) was achieved using 1% plain lidocaine injection. Effective penile block was confirmed by loss of the BCR. Two PVS ejaculation trials were performed: one trial during the penile block and one trial when the penile block had worn off. In 4 subjects, the bladder contents were analyzed for retrograde ejaculation. RESULTS: With the penile block, ejaculation was inhibited in 100% of the subjects. None of the bladder washings demonstrated sperm, indicating absence of retrograde ejaculation. None of the subjects exhibited their usual erectile response, somatic responses, or signs of autonomic dysreflexia. After the penile block wore off, PVS induced ejaculation in all subjects. If subjects usually had erection, somatic responses, or signs of autonomic dysreflexia, these also returned. CONCLUSIONS: Our data suggest that ejaculatory response to PVS in SCI men requires the presence of intact dorsal penile nerves. PMID- 10840109 TI - No-scalpel vasal sperm aspiration and in vitro fertilization for the treatment of anejaculation. AB - OBJECTIVES: To evaluate the utility of no-scalpel vasal sperm aspiration and in vitro fertilization for the treatment of male infertility due to anejaculation. METHODS: Vasal aspiration was performed using a no-scalpel technique, and the sperm were used for conventional in vitro fertilization in 1 patient and intracytoplasmic sperm injection in 6 patients. RESULTS: The average total sperm count was 6 x 10(6), with the motility between 20% and 95%. No complications have been observed to date. A high fertilization rate (74.6%) and pregnancy rate (5 of 7) was achieved with subsequent intracytoplasmic sperm injection or in vitro fertilization. CONCLUSIONS: No-scalpel vasal sperm aspiration is a simple, effective alternative to sperm retrieval, especially for anejaculatory males. PMID- 10840110 TI - Antioxidant activity in the semen of fertile and infertile men. AB - OBJECTIVES: To evaluate catalase- and superoxide dismutase (SOD)-like activities in the seminal plasma of fertile and infertile men. METHODS: Semen samples were obtained from consecutive men presenting for vasectomy (n = 12) and infertility evaluation (n = 105) at our institution. Catalase-like activity was measured by the decrease in hydrogen peroxide after incubation with seminal plasma. SOD-like activity was measured as the inhibition of nitroblue tetrazolium reduction due to superoxide anion generation by xanthine plus xanthine oxidase. RESULTS: Mean seminal catalase-like activity (+/-SEM) in fertile men was not significantly different from that of infertile men (369 +/- 49 versus 326 +/- 17 U/mL, respectively). Mean SOD-like activity in the semen of infertile men was significantly greater than in the semen of fertile controls (46.7 +/- 1.5 versus 37.0 +/- 2.8 U/mL, respectively, P <0.05). CONCLUSIONS: Our data show that infertile men do not have deficient seminal plasma SOD- and/or catalase-like activity (two key antioxidants). These findings suggest that the high semen ROS levels in some infertile men are likely due to excessive generation of ROS rather than deficient ROS scavenging activity in semen. PMID- 10840111 TI - Is a second injection of vasoactive medication necessary during color duplex doppler evaluation of young patients with veno-occlusive erectile dysfunction? AB - OBJECTIVES: To assess the contribution of a second injection of vasoactive medication into the corpus cavernosum during the duplex Doppler study for evaluation of young patients with vasculogenic erectile dysfunction (ED). METHODS: Thirty-five patients, 23 to 50 years old (median age 43), clinically diagnosed as having veno-occlusive ED underwent a color duplex Doppler study. The peak systolic velocity, end-diastolic velocity, resistance index, and arterial diameter measurements were obtained from both cavernosal arteries before and 2, 5, 10, and 20 minutes after an intracavernous injection of a vasoactive drug. All patients were reinjected with similar doses of the same medication, and all measurements were repeated. The erection quality was estimated 20 minutes after each injection. RESULTS: The peak systolic velocity after the second injection was significantly higher statistically than after the first injection (P <0.02). No differences in end-diastolic velocity, resistance index, arterial diameter, or the evaluated side between the first and second injections were found. CONCLUSIONS: A single injection of vasoactive drugs can provide sufficient information on the arterial and veno-occlusive mechanism during color duplex Doppler evaluation of young patients with vasculogenic ED without compromising the quality of the evaluation. Limiting the number of injections to only one may prevent adverse reactions caused by the second injection and reduce the cost and duration of this test. PMID- 10840112 TI - Role of interferon-alpha-2B in prevention of testicular atrophy with unilateral mumps orchitis. AB - OBJECTIVES: To determine the effectiveness of intravenously administered interferon-alpha-2B for the prevention of testicular atrophy in postpubertal men with mumps orchitis. METHODS: Eighteen patients with mumps orchitis were evaluated. The patients were hospitalized, and 3 x 1,000,000 IU intravenous interferon-alpha-2B was administered daily for a total of 7 days. All patients were evaluated by testicular biopsy on the 12th month after interferon treatment. RESULTS: Biopsy results demonstrated total atrophy of the seminiferous tubules in seven (38.8%), 10% atrophy in three (16.6%), and no apparent histopathologic alterations except an arrest in spermatogenesis in eight (44.6%) testes. CONCLUSIONS: Systematic treatment with interferon-alpha-2B does not seem completely effective in preventing testicular atrophy after mumps orchitis. Further investigations involving larger populations are needed. PMID- 10840113 TI - Transient increase in mitogen-induced lymphoproliferative responses in patients with testicular cancer after BEP chemotherapy. AB - OBJECTIVES: To investigate the impact of polychemotherapy on cellular immunity in patients with testicular cancer. METHODS: Lymphocyte subpopulations, lymphoproliferative responses to mitogenic stimulation, and mitogen-induced release of soluble interleukin-2 receptor from peripheral blood mononuclear cells were investigated in 15 patients with testicular germ cell tumors a median of 61 months (range 7 to 73) after polychemotherapy with bleomycin, etoposide, and cisplatin (BEP). RESULTS: The numbers of peripheral blood T cells (CD3+), CD4+ and CD8+ subsets, and lymphoproliferative responses to pokeweed mitogen, phytohemagglutinin, and concanavalin A in patients were comparable to those of healthy control subjects. When two groups of patients were formed according to elapsed time from BEP polychemotherapy and study onset (group A, 12 months and group B, 69 months after termination of BEP), a significant increase in lymphoproliferative response to concanavalin A (P <0.05) was found in group A 1 year after chemotherapy. CONCLUSIONS: BEP chemotherapy administered to patients with testicular cancer does not result in impairment of cellular immunity but rather leads to a significant increase in the capacity of patients' lymphocytes to respond to mitogenic stimulation up to 1 year after polychemotherapy. Moreover, the increased T-cell activity found after BEP therapy may contribute to the high rate of long-term complete remission. PMID- 10840114 TI - Vesicoureteral reflux and videourodynamic studies: results of a prospective study. AB - OBJECTIVES: To study the influence of bladder instability on the conservative management and surgical treatment of children with vesicoureteral reflux (VUR), 102 children were included in a prospective study. METHODS: During a 5-year period all children suspected to have VUR underwent a videourodynamic study to determine VUR grade and bladder function. This resulted in a group of 36 boys and 66 girls who were followed up for well over 5 years. RESULTS: Bladder instability was found in 41 of 102 children (40%). The 102 children were either treated conservatively or surgically. Of the 77 children who were treated conservatively, bladder instability was found in 35 patients. In the conservatively treated group with bladder instability, reflux resolved in 57%; whereas in those with normal bladder function, reflux resolved in 67%. Of the 25 patients who were treated surgically, the operation was successful in 91%. Breakthrough infections occurred in 22 girls and 3 boys, including 14 of 41 patients with bladder instability (34%) and 11 of 61 patients with normal bladder function (18%). CONCLUSIONS: Bladder instability is a frequent finding and an important factor in the treatment of children with VUR. To determine if a patient has VUR and bladder instability a videourodynamic study has proved to be an easy and efficient diagnostic tool. When bladder instability is treated with anticholinergic medication, almost the same results can be expected from conservative treatment as from surgical treatment compared to children with a normal bladder function. Breakthrough urinary tract infections occur more often in girls and tend to occur more often in children with bladder instability. PMID- 10840115 TI - A new tool to aid the urologist in the placement of stents for impacted ureteral stones or strictures: the glide catheter. AB - We describe a technique one can use when only a lubricious wire is able to pass an impacted stone that could otherwise not be passed, over which a new hydrophilic "Glide Catheter is then passed beyond the stone. The lubricious wire can then be exchanged for a stiffer wire that will aid the retrograde passage of a ureteral catheter, stent, or other endourologic device in both a safe and effective manner. PMID- 10840116 TI - Incidental bilateral ureteroceles presenting during investigation for hypertension. PMID- 10840117 TI - An unusual case of impalement injury to the urinary bladder. AB - Isolated impalement injury to the urinary bladder is a rare occurrence. We present a case in which the delay in the diagnosis reveals the potential pitfalls in the management of such injuries. PMID- 10840118 TI - The retained penile prosthesis reservoir: a risk. AB - Complications associated with penile prostheses include malfunction, infection, and patient dissatisfaction. We present a rare complication of a retained abdominal reservoir eroding into the bladder and causing irritative voiding symptoms several years after the removal of the penile components of a three piece penile prosthesis. PMID- 10840119 TI - Primary testicular seminoma in a patient with a history of extragonadal non seminomatous germ cell carcinoma. AB - Extragonadal germ cell carcinoma represents between 3% and 5% of all germ cell carcinomas. A metachronous primary germ cell carcinoma is exceedingly rare in these patients. We report the eighth case, which occurred in a 29-year-old man who presented with testicular seminoma 7 years after his initial presentation with extragonadal non-seminomatous germ cell carcinoma. The seven other patients also presented with extragonadal non-seminomatous germ cell carcinoma, followed subsequently by testicular seminoma in 6 patients and non-seminomatous germ cell carcinoma in the seventh. The mean time to presentation was 8 years. Although rare, this case emphasizes the need for long-term surveillance, including testicular evaluation of patients with a history of extragonadal germ cell carcinoma. PMID- 10840120 TI - Concordant prune belly syndrome in monozygotic twins. AB - We report 2 cases of concordant prune belly syndrome occurring in monozygotic twins. In addition to suggesting a genetic basis for this disease, our 12-year follow-up of these cases illustrates that these patients with an otherwise poor prognosis can have normal growth, development, and renal function with appropriate treatment. PMID- 10840121 TI - Nephrogenic adenomas occurring in a patient with simultaneous kidney-pancreas transplant. AB - We present a female patient who developed multiple nephrogenic adenomas of the bladder about 3 years after a simultaneous kidney-pancreas transplant. The patient was initially treated with endoscopic resection but, due to recurrences, she later underwent a conversion of pancreatic drainage from the bladder to the small bowel and has since done well. We believe this to be the first such case reported in the literature. PMID- 10840122 TI - Primary malignant bladder carcinoid--a diagnostic and management dilemma. AB - A middle-aged man who presented with gross hematuria and irritative voiding symptoms was found to have an invasive infiltrating primary bladder carcinoid after radical cystoprostatectomy. Hurricane chest secondaries were prominent in the follow-up evaluation. The immunohistochemical markers for neuroendocrine differentiation were substantiated. The published reports of primary bladder carcinoid tumors are discussed. PMID- 10840123 TI - Sildenafil citrate-associated priapism. AB - We report the first case of priapism associated with the use of sildenafil citrate. The patient was a 28-year-old man with mild erectile dysfunction after penile trauma who self-treated with a 100-mg dose of sildenafil citrate, resulting in low-flow priapism. After aspiration and intracorporal injection of alpha-agonists, the priapism resolved. The patient's erectile function returned to his prepriapism baseline. PMID- 10840124 TI - Antitumor effects of Scutellariae radix and its components baicalein, baicalin, and wogonin on bladder cancer cell lines. AB - OBJECTIVES: To investigate the antitumor effects of Scutellariae radix and its components baicalein, baicalin, and wogonin on human bladder cancer cell lines (KU-1 and EJ-1) and a murine bladder cancer cell line (MBT-2). METHODS: Bladder cancer cells were incubated with various concentrations of the agents. Antiproliferative activity against the bladder cancer cell lines was examined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diplenyl tetrazolium bromide assay. In an in vivo experiment, the mice were subcutaneously injected with MBT-2 cells, and Scutellariae radix was orally administered at a dose of 2 or 10 mg per mouse one time daily for 10 days from day 11 to day 20. RESULTS: All the drugs inhibited cell proliferation in a dose-dependent manner, but baicalin exhibited the greatest antiproliferative activity. The concentration of baicalin necessary to obtain 50% inhibition was 3.4 microg/mL for KU-1, 4.4 microg/mL for EJ-1, and 0.93 microg/mL for MBT-2. For KU-1 and MBT-2, the percentage of cell survival significantly decreased (P <0.05) at a baicalin concentration of 1 microg/mL. In an in vivo experiment, antitumor effects of Scutellariae radix on C3H/HeN mice implanted with MBT-2 were investigated. All the control mice showed a progressive increase in tumor volume, reaching 2.81 +/- 0.18 cm(3) on day 20 and 5.36 +/- 0.44 cm(3) on day 25. However, when Scutellariae radix was orally administered at a dose of 10 mg per mouse one time daily for 10 days from day 11 to day 20, the tumor volume was 1.99 +/- 0.19 cm(3) on day 20 and 3.86 +/- 0.26 cm(3) on day 25, a significant inhibition of tumor growth (P <0.05). Conclusions. These results suggest that Chinese herbal medicines may become an attractive and promising treatment for bladder cancer. PMID- 10840125 TI - Influences of external urethral sphincter relaxation induced by alpha bungarotoxin, a neuromuscular junction blocking agent, on voiding dysfunction in the rat with spinal cord injury. AB - OBJECTIVES: To determine whether external urethral sphincter (EUS) relaxation induced by alpha-bungarotoxin, a highly selective neuromuscular junction blocking agent, could ameliorate voiding dysfunction after spinal cord injury (SCI) in rats. METHODS: The effects of intravenous alpha-bungarotoxin (333 microg/kg) were evaluated during cystometry in decerebrate, unanesthetized female Sprague-Dawley rats (250 to 300 g) with spinal cords chronically transected at T7-9 (n = 7) or with normal spinal cords (NSC) (n = 7). Parameters measured included voided volume (VV), residual volume (RV), volume threshold for inducing micturition (VT), voiding efficiency (VE), micturition pressure (MP), pressure threshold for inducing micturition (PT), bladder contraction duration (BCD), and compliance (CP). RESULTS: In SCI rats, treatment with alpha-bungarotoxin improved voiding. The toxin increased VE (31%) and reduced RV (42%), MP (52%), BCD (14%), and VT (31%). VV, PT, and CP were not altered. In NSC rats, alpha-bungarotoxin decreased VE (23%), increased RV (63%), and decreased MP (36%), VV (38%), and VT (20%) but did not change BCD and CP. CONCLUSIONS: The results of our study demonstrated that alpha-bungarotoxin improved voiding in SCI rats by reducing urethral outlet resistance. However, in NSC rats, the toxin reduced voiding, probably by suppressing high-frequency phasic sphincter activity, necessary for efficient urine elimination in normal animals. The present results provide further support for the view that drugs that depress striated muscle activity can be useful in the treatment of voiding dysfunction after SCI. PMID- 10840126 TI - Circulating adhesion molecules and neutral endopeptidase enzymuria in patients with urolithiasis and hydronephrosis. AB - OBJECTIVES: To measure the concentrations of adhesion molecules (intercellular adhesion molecule-1 ?ICAM-1, E-selectin, and L-selectin) in the serum of patients with renal stones and patients with hydronephrosis caused by obstructive ureteral stones. Renal tubular enzymes were examined from their urine samples to evaluate whether neutral endopeptidase (NEP) behaved as traditional tubular enzyme markers (N-acetyl-beta-glucosaminidase ?NAG and beta-galactosidase ?beta-GAL) in their disease state. METHODS: Three groups were studied. Group 1 included 15 normal volunteers, group 2 included 12 patients with ureteral stones and ipsilateral hydronephrosis, and group 3 included 17 patients with renal stones in one kidney without hydronephrosis or hydrocalycosis. A single, overnight fasting blood and urine sample was collected from each subject. Serum levels of ICAM-1, E-selectin, and L-selectin were measured by enzyme-linked immunosorbent assay. NEP, NAG, and beta-GAL were measured from urine samples, and the enzyme activities were expressed per gram of creatinine. RESULTS: Serum levels of ICAM-1 were higher in groups 2 and 3 (522 +/- 95 and 329 +/- 42 ng/mL, respectively), but the differences were only significant between group 2 and group 1 (263 +/- 32 ng/mL) and group 2 and group 3 (P <0.05). Serum levels of L-selectin were lower in group 3 and were significantly different when compared with groups 1 and 2 (P <0. 05). The serum levels of E-selectin were not significantly different among these three groups. Urinary levels of NEP were lower in group 2, although the levels of NAG and beta-GAL were more elevated than in group 1. CONCLUSIONS: Serum levels of ICAM-1 were elevated in patients with unilateral hydronephrosis caused by a ureteral stone, and E-selectin and L-selectin levels did not change significantly. These findings suggest that ICAM-1 may play a role in renal immune injury to ureteral obstruction. The urinary NEP values were lower in patients with hydronephrosis caused by ureteral stones, and the traditional lysosomal enzymes were increased. The lower urinary NEP values might suggest impairment of ipsilateral renal function. Renal stones per se resulted in no significant changes in serum adhesion molecule levels, although the levels of L-selectin were significantly decreased. PMID- 10840127 TI - Relation between hippocampal gamma waves and behavioral disturbances induced by phencyclidine and methamphetamine. AB - The relationship between hippocampal electrical activity and behavioral hyperactivity induced by either phencyclidine (PCP) or methamphetamine (MAP) was examined in freely behaving rats. The EEGs at the hippocampal CA1 region were simultaneously recorded with the animal's behavior for 2 h after administration of either PCP or MAP. PCP (10 mg/kg, intraperitoneal (i.p.)) significantly increased locomotor activity including rearing, walking, head-weaving and circling. Spectral analysis of the EEG showed that hippocampal gamma waves (30-70 Hz), but not other frequency bands, were significantly increased from 5 to 120 min after systemic injection of PCP. Inactivation of the medial septum with muscimol, a gamma-aminobutyric acid receptor A agonist, 15 min prior to injection of PCP, suppressed both hippocampal gamma waves and locomotor activity. MAP (1.5 mg/kg, i.p.) also increased locomotor activity for longer than 2 h. During the behavioral hyperactivity induced by MAP, hippocampal EEG showed θ and gamma rhythms that were not significantly different from those during walking before MAP. However, MAP-induced behavioral activity was suppressed by pre-injection of muscimol in the medial septum, which also decreased hippocampal gamma activity. It is suggested that the medial septum plays a role in mediating behavioral disturbances induced by both PCP and MAP through control of the hippocampal electrical activity, and that hippocampal gamma waves may play a permissible role in the expression of behaviors. PMID- 10840128 TI - Electrophysiological correlates of cross-linguistic speech perception in native English speakers. AB - The present study examined the electrophysiological responses that Native English speakers display during a passive oddball task when they are presented with different types of syllabic contrasts, namely a labial /ba/-dental /d a/, a Hindi dental /d a/-retroflex /da/ and a within-category (two /ba/ tokens) contrasts. The analyses of the event-related potentials obtained showed that subjects pre attentively perceive the differences in all experimental conditions, despite not showing such detection behaviourally in the Hindi and within-category conditions. These results support the notion that there is no permanent loss of the initial perceptual abilities that humans have as infants, but that there is an important neural reorganisation which allows the system to overcome the differences detected and only be aware of contrasts that are relevant in the language which will become the subjects native tongue. We also report order asymmetries in the ERP responses and suggest that the percepts and not only the physical attributes of the stimuli have to be considered for the evaluation of the responses obtained. PMID- 10840129 TI - Circadian rhythms, aging and memory. AB - In human beings and animal models, cognitive performance is often impaired in natural and experimental situations where circadian rhythms are disrupted. This includes a general decline in cognitive ability and fragmentation of behavioural rhythms in the aging population of numerous species. There is some evidence that rhythm disruption may lead directly to cognitive impairment; however, this causal link has not been made for effects due to aging. We have tested this link by examining rhythms and performance on contextual conditioning with the conditioned place preference task, in elderly, age-matched hamsters. Young healthy hamsters developed a preference for a context that is paired with the opportunity to engage in wheel-running (experiment 1). Aged animals with consolidated locomotor rhythms developed similar degrees of preference, whereas the age-matched hamsters with fragmented rhythms did not (experiment 2). The degree of preference was also correlated with activity amplitude. These results support the notion that age related rhythm fragmentation contributes to the age-related memory decline. PMID- 10840130 TI - Dopamine release in the rat globus pallidus characterised by in vivo microdialysis. AB - Brain microdialysis has been used to examine the in vivo effects of potassium and calcium on dopamine release in the dorsal globus pallidus (GP) of rats. Furthermore, the effects of food presentation and consumption on dopamine release in the GP were investigated. Basal dopamine levels in the GP were below the detection limit, therefore nomifensine (30 microM) was added to the perfused artificial cerebrospinal fluid (aCSF). A prominent increase of dopamine release to 370% was observed after perfusion with elevated potassium (100 mM), while perfusion with calcium-free aCSF produced a significant decrease of dopamine efflux to 36% of control levels. Furthermore, presentation and consumption of food resulted in a rapid increase of extracellular dopamine to 130%. The present experiments demonstrate that in the GP extracellular dopamine can be measured by in vivo brain microdialysis. The data suggest that the dopamine release in the GP can be stimulated by a depolarising agent and involves a partially calcium dependent release mechanism. The data further suggest that dopamine in basal ganglia structures downstream the striatum as the GP is involved in signalling of important stimuli in the environment, e.g. food. PMID- 10840131 TI - Motor learning and Parkinson's disease: refinement of within-limb and between limb coordination as a result of practice. AB - Even though the basal ganglia have been assigned a role in motor learning, few studies have addressed motor learning capabilities in Parkinson patients. In the present experiment, improvement of bimanual figure drawing across practice was compared between Parkinson patients and normal age-matched controls. At regular intervals during acquisition, performance was assessed under normal vision and blindfolded conditions. At initiation of practice, the typical signs associated with Parkinson's disease became evident, such as bradykinesia and hypometria. Moreover, reduced synchronization between the force-time specifications of both limbs was observed. When vision was withdrawn, Parkinson patients showed a larger drift of drawing performance across the workspace, indicative of a decline in proprioception. In spite of the aforementioned deficits, Parkinson patients made marked improvements in the speed of execution, the consistency of the spatial trajectories, and the synchronization between the limbs across practice, even though they never reached the performance levels obtained in elderly controls. The findings demonstrate that Parkinson patients do benefit from practice to refine their upper limb control and to alleviate their most basic motor deficits. PMID- 10840132 TI - Perceptual alternation in obsessive compulsive disorder--implications for a role of the cortico-striatal circuitry in mediating awareness. AB - Mounting evidence suggests that obsessive compulsive disorder (OCD) results from functional aberrations of the fronto-striatal circuitry. However, empirical studies of the behavioral manifestations of OCD have been relatively lacking. The present study employs a behavioral task that allows a quantitative measure of how alternative percepts are formed from one moment to another, a process mimicking the brain state in which different thoughts and imageries compete for access to awareness. Eighteen patients with OCD, 12 with generalized anxiety disorder, and 18 normal subjects participated in the experiment, in which they viewed one of the three Schroder staircases and responded by pressing a key to each perceptual reversal. The results demonstrate that the patients with OCD have a higher perceptual alternation rate than the normal controls. Moreover, the frequency of perceptual alternation is significantly correlated with the Yale-Brown obsessive compulsive and the Hamilton anxiety scores. The increase in the frequency of perceptual reversals cannot easily be accounted for by learning or by different patterns of eye fixations on the task. These results provide further evidence that an impairment of the inhibitory function of the cortico-striatal circuitry might underlie the etiology of OCD. The implications of the results for a general role of the cortico-striatal circuitry in mediating awareness are discussed. PMID- 10840133 TI - Hemispheric interaction, metacontrol, and mnemonic processing in split-brain macaques. AB - These experiments explored the interactions remaining between the cerebral hemispheres in two split-brain macaques. The 'split' was earlier confirmed by showing that one hemisphere was incapable of identifying visual images seen by the other. The critical tests for residual interactions were intermingled with control trials in a continuous recognition task. These tests were of two kinds: 'parallel processing', to determine how simultaneous viewing by both hemispheres affected subsequent recognition by one of them alone; and 'conflict', where opposite responses were demanded from the two hemispheres, thus assessing the issue of metacontrol. Two types of stimuli were also employed: ART, in which each hemisphere saw essentially the same image; and BIPARTITE, in which images were entirely different for each hemisphere. Since, with either type of stimulus, performance was best when viewed by both hemispheres at both encoding and retrieval, 'parallel processing' was highly efficient. However, when both hemispheres viewed initially and only one was subsequently queried, performance was significantly worse than when each hemisphere acted alone on each occasion. It is thus reasoned that when both hemisphere view together, the resultant memory trace somehow reflects the bilaterality, a conclusion concordant with observations of Marcel on blindsight. Processing different images (BIPARTITE) was somewhat more disruptive in this regard than if the same image was viewed by each hemisphere. This was particularly true in the conflict situation, where for one hemisphere the item seen was NEW and for the other it was OLD. A response of 'OLD' was, at first, consistently rewarded. When this well-established protocol was changed, the hemispheres in each animal were gradually able to revise their joint behavior. This, together with the effect of disparate images, and the deficiency evoked when the animals were forced to recognize unilaterally an image first viewed under bilateral conditions, all manifest considerable, and complex, interaction between the hemispheres despite absence of the forebrain commissures. The superior colliculus seems a likely focal point for such interhemispheric effects. PMID- 10840134 TI - Stress induces rapid changes in serotonergic activity: restraint and exertion. AB - Rapid activation of central serotonergic systems occurs in response to the social stress of aggression in dominant lizards. The most rapid expression of serotonergic activity occurs in nucleus accumbens, hippocampus and brainstem. To compare previously measured responses induced by social stressors with those provoked by physical stress, serotonergic activity was examined following restraint stress (handling) and forced physical exertion. After handling, some male Anolis carolinensis were placed on a race track and either run until there was no movement following 1 min of prodding, or half that time. Controls were killed without treatment. Lizards stressed by handling showed rapid (25 s) increases in serotonergic activity (5-HIAA/5-HT) in striatum, dorsal cortex, locus ceruleus, and nucleus accumbens. Other changes in serotonergic systems caused by stress occurred in raphe and hippocampus. Serotonergic changes induced by handling stress were reversed by exercise (to 50% maximal exertion time) in subiculum, striatum and nucleus accumbens. The serotonergic profile of lizards run until they would no longer respond to prodding (maximal exertion time) was significantly different from that for more acute exertion in hippocampus, subiculum, striatum, medial amygdala, locus ceruleus, area postrema, and raphe. Physical stress (handling) mimicked social stress by producing rapid serotonergic changes in hippocampus, subiculum, nucleus accumbens and locus ceruleus. In contrast, the medial amygdala, which has previously been demonstrated to respond serotonergically to social stress only after a temporal delay, did not show a rapid response to restraint stress. PMID- 10840135 TI - Different hippocampal molecular requirements for short- and long-term retrieval of one-trial avoidance learning. AB - Rats were trained in one-trial step-down inhibitory avoidance and tested either 3 h or 31 days later. Ten minutes prior to the retention test, through indwelling cannulae placed in the CA1 region of the dorsal hippocampus, they received 0.5 microl infusions of: saline, a vehicle (2% dimethylsulfoxide in saline), the glutamate NMDA receptor blocker, aminophosphonopentanoic acid (AP5) (5.0 microg), the AMPA/kainate receptor blocker, cyanonitroquinoxaline dione (CNQX) (0.25 or 1.25 microg), the metabotropic receptor antagonist, methylcarboxyphenylglycine (MCPG) (0.5 or 2.5 microg), the inhibitor of calcium/calmodulin-dependent protein kinase II (KN62) (3.5 microg), the inhibitor of cAMP-dependent protein kinase (PKA), Rp-cAMPs (0.1 or 0.5 microg), the stimulant of the same enzyme, Sp-cAMPs (0.1 or 0.5 microg), or the inhibitor of the mitogen-activated protein kinase (MAPK) kinase, PD098059 (10 or 50 microM). CNQX, KN62 and PD098059 were dissolved in the vehicle; the other drugs were dissolved in saline. All these drugs, at the same doses, had been previously found to affect short- and long-term memory formation of this task. Retrieval measured 3 h after training (short-term memory) was blocked by CNQX and MCPG, and was unaffected by all the other drugs. In contrast, retrieval measured at 31 days was blocked by MCPG, Rp-cAMPs and PD098059, enhanced by Sp-cAMPs, and unaffected by CNQX, AP5 or KN62. The results indicate that, in CA1, glutamate metabotropic receptors are necessary for the retrieval of both short- and long-term memory; AMPA/kainate receptors are necessary for short-term but not long-term memory retrieval, and NMDA receptors are uninvolved in retrieval. Both the PKA and MAPK signalling pathways are required for the retrieval of long-term but not short-term memory. PMID- 10840136 TI - Pituitary-adrenal axis and behavioural responses of maternally deprived juvenile rats to the open field. AB - Adult rats submitted to maternal deprivation (DEP) on post-natal day (pnd) 11 show smaller corticosterone (CORT) response to a saline injection than non deprived (NDEP) rats, mainly at 30 days of age. In the present study we sought to investigate the pituitary-adrenal axis response of 30-day-old DEP rats to a 5-min open-field session, with lights and sound on. Hormone levels were assessed immediately or 20 min after the end of the stress, and were compared to basal levels. The immediate ACTH response of DEP females was the highest; this difference was no longer observed at 20 min, but hormone levels were still higher than basal. Both males and females showed an augmented CORT secretion immediately after the open field session, although only the response of NDEP females was higher than that of male counterparts. Moreover, DEP females showed a lower CORT response than their NDEP counterparts immediately after the stress. The CORT stress response remained equally elevated in males and females at the 20-min time point. Finally, DEP animals, regardless of the gender, ambulated more in the centre of the open field and displayed less grooming behaviour than NDEP pups, suggesting that DEP rats are less emotional than NDEP animals. PMID- 10840137 TI - Specificity of memory measures in an adjusting delayed nonmatching-to-position task for rats. AB - While the delayed nonmatching-to-position (DNMTP) behavioral paradigm has often been used by neuroscientists to assess working memory in rats, its measure of working memory is compromised by floor and ceiling effects. Specifically, these floor and ceiling effects undermine the ability to detect a significant interaction in a two-factor repeated-measures ANOVA, which is required in order to conclude that impaired performance has resulted from disrupted working memory and not from a change in another psychological process (i.e. that the impairment is 'specific' to working memory). The present study was conducted to evaluate if these limitations could be overcome in a DNMTP by adjusting the length of time that the rat was required to remember (the 'delay') so as to avoid the floor and ceiling. The general procedure for two experiments presented subjects with trials where there was either a minimal (1 s) delay or a longer delay of varying length, with the goal of maintaining nonmatch-to-position accuracy near 75%. The procedure was such, that if the average accuracy was at or above 75%, then the next trial would be a long-delay trial. If it were below 75%, then the next trial would be a 1 s delay trial. In the first experiment, the subjects were presented with trials where the value of the longer delay was systematically varied between sessions. This was done to simulate the faster rate of forgetting found in persons with amnesia. DNMTP accuracy diminished at the longer delay but not the short delay as the second interval was lengthened. However, other measures, including accuracy at the 1 s delay, discrimination accuracy during the sample phase, and the number of trials completed per session, did not change. This experiment suggested that this DNMTP could precisely measure small changes in the rate of forgetting. In a second experiment, a potential non-mnemonic confound, the level of motivation, was directly manipulated by giving the water-restricted subjects access to water immediately prior to the start of the session. The number of trials completed per session diminished, but the accuracy at both the 1 s and the longer delays did not change. These results indicated that the measures of working memory in this DNMTP were insensitive to changes in motivation. Together, these experiments confirmed that adjusting the delay in the DNMTP improves the validity of the measures in this task. PMID- 10840138 TI - Neuropeptide Y in brains of the Flinders Sensitive Line rat, a model of depression. Effects of electroconvulsive stimuli and d-amphetamine on peptide concentrations and locomotion. AB - Neuropeptide Y (NPY), has been implicated in the pathophysiology of depression and the mechanisms of action of electroconvulsive treatment (ECT). In this series of experiments, we explored whether there are differences between Flinders Sensitive Line (FSL) rats, an animal model of depression, and controls, Flinders Resistant Line (FRL) in (1) baseline brain NPY-LI concentrations, (2) effects of ECS on locomotion and brain neuropeptides, (3) amphetamine effects on behavior, and (4) effects of ECS pretreatment on subsequent effects of amphetamine on behavior. Both strains were divided into two groups, receiving eight ECS or ShamECS. Twenty-four hours after the last session, animals were habituated in activity boxes for 45 min before given d-amphetamine (1.5 mg.kg(-1), subcutaneously) or vehicle. Locomotor activity was then recorded for an additional 45 min. Twenty-four hours later, rats were sacrificed by microwave irradiation, the brains dissected into frontal cortex, occipital cortex, hippocampus, hypothalamus and striatum, and the neuropeptides extracted and measured by radioimmunoassay. No differences between FSL and FRL rats in baseline locomotor activity were found. FSL compared to FRL animals showed a significantly larger locomotion increase following saline and a significantly smaller increase following amphetamine. ECS pretreatment significantly decreased the saline effects on locomotion in the FSL and the amphetamine effects in the FRL rats. 'Baseline' NPY-like immunoreactivity (LI) concentrations were lower in the hippocampus of the 'depressed' rats. ECS increased NPY-LI in frontal cortex, occipital cortex and hippocampus of both strains. The hippocampal NPY-LI increase was significantly larger in the FSL compared to FRL animals. PMID- 10840139 TI - Individual and combined manipulation of muscarinic, NMDA, and benzodiazepine receptor activity in the water maze task: implications for a rat model of Alzheimer dementia. AB - Recent evidence indicates that Alzheimer disease typically involves different degrees of impairment in a variety of neurotransmitter systems, behaviors, and cognitive abilities in different patients. To investigate the relations between neurotransmitter system, behavioral, and cognitive impairments in an animal model of Alzheimer disease we studied spatial learning in a Morris water maze in male Long-Evans rats given neurochemical agents that targeted muscarinic cholinergic, NMDA, or benzodiazepine systems. Naive rats given a single agent or a combination of agents were severely impaired in place responding and had behavioral strategy impairments. Rats made familiar with the required water maze behavioral strategies by non-spatial pretraining performed as well as controls if given a single agent. Non-spatially pretrained rats with manipulation of both muscarinic cholinergic and NMDA or muscarinic cholinergic and benzodiazepine systems had a specific place response impairment but no behavioral strategy impairments. The results suggest that impairment of both muscarinic cholinergic and NMDA, or muscarinic cholinergic and benzodiazepine systems may model some aspects of human Alzheimer disease (impairments in navigation in familiar environments), but not other aspects of this disorder (global dementia leading to general loss of adaptive behavior). Previous research suggests that impairment of both muscarinic cholinergic and serotonergic systems may provide a better model of global dementia. The water maze testing and detailed behavioral analysis techniques used here appear to provide a means of investigating the contributions of various combinations of neurotransmitter system impairments to an animal model of Alzheimer disease. PMID- 10840140 TI - Unilateral lesions of the medial agranular cortex impair responding on a lateralised reaction time task. AB - The present study assessed the behavioural sequalae of unilateral excitotoxic cortical lesions made either in the medial agranular cortex (AGm) or in the medial prefrontal cortex (mPFC) using a visual reaction time task. The task required animals to sustain a nose-poke in a central hole, until a brief light stimulus was presented in either of two holes which were located on the same side of the box: this enabled performance on each side of the rat's body to be assessed independently. Lesions of the AGm impaired performance on the contralateral side, with rats biasing their responding to the nearer of the two response locations. Analysis of the deficit revealed that rats were able to discriminate between the two stimuli and suggested that AGm lesions disrupted the control of contralateral responding. Lesions of the mPFC produced similar response-related deficits, but these were more transient in nature. Neither AGm lesions nor mPFC lesions impaired performance on the ipsilateral side, consistent with the concept of an egocentrically coded deficit. PMID- 10840141 TI - Unconditioned anxiety and social behaviour in two rat lines selectively bred for high and low anxiety-related behaviour. AB - Individuals of high anxiety-related behaviour (HAB) and low anxiety-related behaviour (LAB) rat lines were selectively bred for differences in anxiety related behaviour on the elevated plus-maze. The goal of this study was to investigate whether this behavioural difference is restricted to the test used as the selection criterion or whether it is a stable and robust trait also in other conflict or non-conflict situations. Therefore, behaviour of male and female HAB and LAB rats was examined in two further tests of unconditioned anxiety: the black-white box and the social interaction test. Furthermore, behaviour of group housed male HAB and LAB rats was studied in their home cages. In addition to standard statistics, discriminant analyses were performed. The difference in anxiety-related behaviour between the two lines was highly consistent in all tests of unconditioned anxiety. There were also differences in home cage behaviour, LAB rats being more active than HAB rats; this is likely to be a consequence of the LAB rats displaying a higher aggressiveness in social behaviour, compared to HAB rats. In all tests used HAB and LAB rats were clearly distinguished by discriminant analysis. However, while in the elevated plus-maze and the black-white box test the most important parameters for discrimination between the two lines were mainly those generally seen as closely related to anxiety, the discrimination in the social interaction paradigm was primarily due to differences in locomotor activity. PMID- 10840142 TI - Postnatal intracerebroventricular administrations of NGF alter spatial memory in adulthood. AB - The present work assessed the effects of intracerebroventricular injections (2x5 mg/2.5 ml) of recombined human nerve growth factor (rhNGF) at postnatal days 2 and 3 upon the development of spatial learning capacities in rats. The treated rats were trained at the age of 22 days to escape onto an invisible platform at a fixed position in space in a Morris navigation task. For half of the subjects, the training position was also cued, a procedure aimed at facilitating escape and reducing attention to the distant spatial cues. At the age of 2 months all the rats were retrained in the same task. Treatment effects were found in both immature and adult rats. The injection of NGF induced a slight alteration of the immature rats' performance. In contrast, a marked impairment of spatial abilities was shown in the 2-month-old rats. The most consistent effects were a significant increase in the escape latency and a decrease bias towards the training platform area during probe trials. The reduction of spatial memory was particularly marked if the subjects had been trained in a cued condition. Taken together, these experiments reveal that an acute pharmacological treatment that leads to transient modifications during early development might induce a behavioural change long after treatment. Thus, the development and the maintenance of an accurate spatial representation are tightly related to the development of brain structures that could be altered by precocious NGF administrations. PMID- 10840143 TI - L-type calcium channels selectively control the defensive behaviors induced by electrical stimulation of dorsal periaqueductal gray and overlying collicular layers. AB - The present study reports the involvement of L-type calcium channels in the control of defensive behaviors produced by electrical stimulation of dorsal periaqueductal gray and overlying collicular layers. Rats that had chemitrodes in the dorsal midbrain and which stimulation produced freezing or flight behaviors with less than 55 microA were selected for drug experiments. Stimulation was repeated the day after the screening session 20 min following the microinjection into the dorsal periaqueductal gray of 15 nmol of either verapamil, a selective L type calcium channel antagonist, or cobalt chloride (CoCl(2)), a calcium-specific channel modulator. Post-drug sessions were performed 48 h after. Threshold functions were obtained by logistic fitting of accumulated response frequencies. Verapamil and CoCl(2) significantly attenuated the output of immobility, exophthalmus, running and jumping. Although to a lesser degree, verapamil also attenuated defecation. Because CoCl(2) had no effect on defecation, the attenuation of this response by verapamil suggests a non-specific action of this drug. Neither verapamil nor CoCl(2) changed the output of micturition. Finally, whereas there was a complete recovery of defensive thresholds following the microinjection of verapamil, the attenuating effects of CoCl(2) were still present 48 h after. These results support an important role of L-type calcium channels in the neurogenesis of dorsal periaqueductal gray-evoked immobility, exophthalmus, running and jumping, but not defecation and micturition responses. PMID- 10840144 TI - Distinct patterns of behavioural impairments resulting from fornix transection or neurotoxic lesions of the perirhinal and postrhinal cortices in the rat. AB - The present study provides evidence that lesions of the fornix (FNX) and of the perirhinal/postrhinal cortex (PPRH), which both disconnect the hippocampus from other brain regions, can lead to distinct patterns of behavioural impairments on tests of spatial memory and spontaneous object recognition. For example, whereas FNX lesions impaired allocentric spatial delayed alternation in a T-maze but generally spared a test of spontaneous object recognition, PPRH lesions produced the opposite pattern of results. Indeed, on the T-maze task PPRH animals significantly outperformed controls when the retention delay was increased to 60 s. In addition, some evidence was found that contributions from both the fornix and perirhinal/postrhinal cortex may be required when object and spatial information must be integrated. In an object-in-place test, for example, PPRH animals failed according to two measures, and FNX animals failed according to one measure, to discriminate objects that had remained in fixed locations from those that had exchanged locations with other objects. Neither lesion, however, affected performance of a visuospatial conditional task, a Pavlovian autoshaping task, or a one-pair pattern discrimination task. It is suggested that the perirhinal/postrhinal cortex, rather than being specialised for a particular type of associative learning, is important for processing complex visual stimuli. PMID- 10840145 TI - Withdrawal-associated changes in peripheral nitrogen oxides and striatal cyclic GMP after chronic haloperidol treatment. AB - The irreversible nature of haloperidol-induced tardive dyskinesia suggests a neurotoxic etiology, although the causes are unknown. Since nitric oxide demonstrates neurotoxic as well as neuroprotectant properties, and antipsychotics can inhibit nitric oxide (NO) synthase in vitro, this study investigates the NO cGMP pathway as a pre-determining factor in chronic haloperidol-associated dyskinesia in rats. Sprague-Dawley rats were administered either water, oral haloperidol (0.25 mg/kg per day po), the guanylyl cyclase-nNOS inhibitor, methylene blue (MB; 5 mg/kg per day ip) or haloperidol plus MB for 3 weeks. In a second protocol, rats received water or haloperidol orally for 17 weeks, followed by 3 weeks withdrawal. Either saline (ip) or MB (ip) was administered for 3 weeks prior to haloperidol withdrawal. Vacous chewing movements (VCMs) were continuously monitored, followed by the determination of serum nitrogen oxides (NO(x)) and striatal cGMP at week 20. Chronic haloperidol engendered significant VCMs, with acute withdrawal resulting in significantly reduced plasma NO(x) and striatal cGMP. Furthermore, NO(x) and cGMP suppression was amplified by pre withdrawal MB administration. Sub-acute haloperidol similarly induced incremental VCMs, but without effect on NO(x) or cGMP. However, haloperidol plus MB also induced significantly greater VCMs with decreased cGMP compared to haloperidol alone. Thus, NO(x)-cGMP inhibition persists pronounced after long-term haloperidol treatment and withdrawal. MB potentiation of these effects suggests that haloperidol inhibits a NO-dependent neuro-protective response to oxidative stress in the striatum that may pre-determine TD development. PMID- 10840146 TI - Conditional discrimination learning in rats with global ischaemic brain damage. AB - Hippocampal cell loss was induced by the four-vessel occlusion (4VO) method, a model of global ischaemia. Global ischaemia for 15 min induced a selective damage to the CA1 subfield. Occlusion for 25 min produced a larger cell loss within the CA1 and more variably the CA2, CA3, the striatum and cortex. Ischaemic and sham control groups were assessed on two conditional discrimination tasks (presenting the conditional cues either in the choice arms or the start arm) and two spatial tasks (water maze and a simple spatial discrimination task). No significant effects were found on either of the spatial tasks (apart from the speed measure on the water maze). However, on the conditional discrimination task with the cues in the choice arms, animals with 25 min ischaemia learned the task significantly more slowly than the 15 min ischaemic and control groups. Results for the task with cues presented in the start arm differed according to choice of criterion for learning. With a standard criterion of 90% accuracy on one session controls were significantly superior to both ischaemic groups. However, in this task rats with 15 min occlusion showed the greatest impairment, and were significantly worse than both the controls and the 25 min occlusion group. These results suggest that hippocampal ischaemic damage disrupts the learning of conditional discrimination but not simple spatial tasks. No clear relationship between the extent of hippocampal cell loss and behavioural impairment was evident. These results highlight the critical importance of procedural factors in the assessment of cognitive impairment. PMID- 10840147 TI - Effects of transcutaneous electrical nerve stimulation (TENS) on cognition and behaviour in aging. AB - In previous studies, transcutaneous electrical nerve stimulation (TENS) improved cognition and behaviour in patients with Alzheimer's disease (AD). The rationale underlying these studies was that TENS could activate, e.g. the septo-hippocampal region and the hypothalamus through direct and indirect pathways. As these areas are also affected in normal aging, the present study examined the effects of TENS on cognition and behaviour in nondemented elderly persons. The results suggest an improvement in visual short-term and verbal long-term (recognition) memory, and semantic verbal fluency. Moreover, stimulated elderly persons felt less depressed. Limitations are discussed. PMID- 10840148 TI - Mechanisms underlying hypoxia-induced neuronal apoptosis. AB - In vivo models of cerebral hypoxia-ischemia have shown that neuronal death may occur via necrosis or apoptosis. Necrosis is, in general, a rapidly occurring form of cell death that has been attributed, in part, to alterations in ionic homeostasis. In contrast, apoptosis is a delayed form of cell death that occurs as the result of activation of a genetic program. In the past decade, we have learned considerably about the mechanisms underlying apoptotic neuronal death following cerebral hypoxia-ischemia. With this growth in knowledge, we are coming to the realization that apoptosis and necrosis, although morphologically distinct, are likely part of a continuum of cell death with similar operative mechanisms. For example, following hypoxia-ischemia, excitatory amino acid release and alterations in ionic homeostasis contribute to both necrotic and apoptotic neuronal death. However, apoptosis is distinguished from necrosis in that gene activation is the predominant mechanism regulating cell survival. Following hypoxic-ischemic episodes in the brain, genes that promote as well as inhibit apoptosis are activated. It is the balance in the expression of pro- and anti-apoptotic genes that likely determines the fate of neurons exposed to hypoxia. The balance in expression of pro- and anti-apoptotic genes may also account for the regional differences in vulnerability to hypoxic insults. In this review, we will examine the known mechanisms underlying apoptosis in neurons exposed to hypoxia and hypoxia-ischemia. PMID- 10840149 TI - Evidence for transcortical reflex pathways in the lower limb of man. AB - The existence of transcortical reflex pathways in the control of distal arm and hand muscles in man is now widely accepted. Much more controversy exists regarding a possible contribution of such reflexes to the control of leg muscles. It is often assumed that transcortical reflex pathways play no, or only a minor, role in the control of leg muscles. Transcortical reflex pathways according to this view are reserved for the control of the distal upper limb and are seen in close relation to the evolution of the primate hand. Here we review data, which provide evidence that transcortical reflexes do exist for lower limb muscles and may play a significant role in the control of at least some of these muscles. This evidence is based on animal research, recent experiments combining transcranial magnetic stimulation with peripheral electrical and mechanical stimulation in healthy subjects and neurological patients. We propose that afferent activity from muscle and skin may play a role in the regulation of bipedal gait through transcortical pathways. PMID- 10840150 TI - Implication of cysteine proteases calpain, cathepsin and caspase in ischemic neuronal death of primates. AB - Although more than 8000 papers of apoptosis are published annually, there are very few reports concerning necrosis in the past few years. A number of recent studies using lower species animals have suggested that the cornu Ammonis (CA) 1 neuronal death after brief global cerebral ischemia occurs by apoptosis, an active and genetically controlled cell suicide process. However, the studies of monkeys and humans rather support necrosis, the calpain-mediated release of lysosomal enzyme cathepsin after ischemia conceivably contributes to the cell degeneration of CA1 neurons. This paper provides an overview of recent developments in ischemic neuronal death, presents the cascade of the primate neuronal death with particular attentions to the cysteine proteases, and also indicates selective cathepsin inhibitors as a novel neuroprotectant. Furthermore, the possible interaction of calpain, cathepsin, and caspase in the cascade of ischemic neuronal death is discussed. PMID- 10840151 TI - Neuromedin B. AB - Neuromedin B (NMB) is one of the bombesin (BN)-related peptides in mammals. It was originally purified from pig spinal cords, and it has been shown to be present in central nervous system as well as in gastrointestinal tract. BN and its related peptides have various physiological effects. These include regulation of exocrine and endocrine secretions, smooth muscle contraction, feeding, blood pressure, blood glucose, body temperature and cell growth. NMB exerts its effect by binding to the cell surface receptor. A high affinity receptor, NMB receptor (NMB-R) has been identified. This is a G-protein coupled receptor with seven membrane-spanning regions. Upon agonist binding, several intracellular signaling cascades including phospholipase activation, calcium mobilization and protein kinase C (PKC) activation lead to expression of several genes, DNA synthesis or cellular effects such as secretion. Existence of NMB-R has been demonstrated in several brain regions, notably in olfactory and thalamic regions, and in gastrointestinal tracts. Recent analysis using NMB-R-deficient mice, generated by gene-targeting technique, enables to distinguish functional properties of NMB-R from GRP-R. In this review, molecular characterization, anatomical distribution and pharmacological properties of NMB and NMB-R will be presented. Moreover, physiological roles of NMB and its receptor demonstrated by the analysis of NMB-R deficient mice will be reported. Comparison with GRP/GRP-R system will provide important information about BN-like peptide systems in mammals. PMID- 10840152 TI - Molecular mechanisms of recovery from vestibular damage in mammals: recent advances. AB - The aim of this review is to summarise and critically evaluate studies of vestibular compensation published over the last 2 years, with emphasis on those concerned with the molecular mechanisms of this process of lesion-induced plasticity. Recent studies of vestibular compensation have confirmed and extended the previous findings that: (i) compensation of the static ocular motor and postural symptoms occurs relatively rapidly and completely compared to the dynamic symptoms, many of which either do not compensate substantially or else compensate variably due to sensory substitution and the development of sensori motor strategies which suppress or minimize symptoms; (ii) static compensation is associated with, and may be at least partially caused by a substantial recovery of resting activity in the ipsilateral vestibular nucleus complex (VNC), which starts to develop very quickly following the unilateral vestibular deafferentation (UVD) but does not correlate perfectly with the development of some aspects of static compensation (e.g., postural compensation); and (iii) many complex biochemical changes are occurring in the VNC, cerebellum and even areas of the central nervous system like the hippocampus, following UVD. However, despite many recent studies which suggest the importance of excitatory amino acid receptors such as the N-methyl-D-aspartate receptor, expression of immediate early gene proteins, glucocorticoids, neurotrophins and nitric oxide in the vestibular compensation process, how these various factors are linked and which of them may have a causal relationship with the physiological changes underlying compensation, remains to be determined. PMID- 10840153 TI - p53 expression in fallopian tube carcinomas. AB - Sixty-three women treated for primary carcinoma of the fallopian tube (PFTC) from 1980-1995 were retrospectively analyzed to study the impact of p53 expression on survival in primary carcinoma of the fallopian tube. The mean age of the patients was 61.2 years (range 37.3-80.2). Twenty-four (38%) patients were FIGO stage I, 11 (18%) stage II, 19 (30%) stage III and nine (14%) stage IV. Complete radical resection was achieved in 45 (71%) patients. In 56 (89%) women, surgery involved removal of the uterus, the adnexa, and/or the omentum or lymph nodes. Adjuvant therapy consisted of either chemotherapy (n: 31; 49%) or irradiation (n: 21; 33%). The 5-year survival rate for all cases was 43%. For stages I+II and III+IV the 5-year survival rate was 59 and 19%, respectively (P<0.00001). Twelve samples (19%) were p53-negative (tumours with <10% of nuclear staining) and 51 (81%) samples were p53 positive tumours with >10% of nuclear staining. The median survival for the p53-negative group was 40 and 21 months for the p53 positive group. No statistical significance between p53 expression and different FIGO stages was observed, however, a trend for a slightly better survival for the p53 negative group was observed. PMID- 10840154 TI - Inverse correlation between KAI1 mRNA levels and invasive behaviour in bladder cancer cell lines. AB - We have previously shown that levels of KAI1 mRNA are dramatically reduced in invasive human bladder cancers. To further investigate the role of KAI1 in bladder cancer, we have examined the relationship between KAI1 mRNA levels and cell behaviour in 18 bladder cancer cell lines and a virus-transformed uro epithelial cell line. We found that low KAI1 mRNA levels correlated with increased in vitro invasive ability, reduced Ca(2+)-dependent and -independent cell-cell adhesion and reduced adhesion to fibronectin. These data support the idea that loss of KAI1 expression is an important factor in tumour cell invasive behaviour. PMID- 10840155 TI - Chromosomal mapping of human genes by radioactive hybridization of cDNAs to Centre d'Etude du Polymorphisme humain high density gridded filter sets. AB - Chromosomal assignment of human transcribed sequences has been done mainly by high throughput genome analysis in specialized genome centres and, in a more classical fashion, by fluorescence in-site hybridization (FISH) analysis. Not every laboratory has the ability to map cDNAs by FISH analysis. We here report a rapid mapping approach that is based on the hybridization of cDNA probes to high density gridded Centre d'Etude du Polymorphisme Humain filters followed by subsequent computational analysis by database searches in the internet. Not only transcribed sequences but also genomic DNA could be subjected to this mapping approach. The presented approach allows to map human transcribed and genomic DNAs within 1-3 days and with a high level of resolution that will constantly increase in line with the incorporation of data deriving from high throughput genome mapping. PMID- 10840156 TI - Peroxiredoxin I expression in oral cancer: a potential new tumor marker. AB - This study investigates the applicability of the novel antioxidant protein, peroxiredoxin (Prx) I as a marker for tumor status in oral squamous cell carcinoma (SCC). Samples from 53 patients with SCC in the oral cavity were examined by immunohistochemistry. Correlations between the expression level of Prx I and proliferating cell nuclear antigen (PCNA), the clinical features of tumors, and their histopathological classifications were statistically analyzed. Cases exhibiting low Prx I expression level included significantly more with larger tumor mass cases (T-category, P=0.004), positive lymph node metastasis (N category, P=0.015), advanced stage (P=0.002), and poorly differentiated cells (P=0.020). There was no significant difference between Prx I expression and the other indices. PMID- 10840157 TI - Effects of pinostrobin on estrogen metabolism and estrogen receptor transactivation. AB - The interaction between the estrogen receptor and 5-hydroxy-7-methoxyflavanone (pinostrobin) was studied in the presence or absence of estradiol or dehydroepiandrosterone sulfate (DHEAS), respectively, using a stably transfected human breast cancer cell line (MVLN). We also evaluated its action on the proliferation in estrogen-dependent (MCF-7) human breast cancer cells in the same conditions than the estrogen receptor assay. On the other hand pinostrobin was evaluated for their effects on the human placental aromatase, 3beta hydroxysteroid dehydrogenase Delta(4)/Delta(5) isomerase and 17beta hydroxysteroid dehydrogenase activities. Pinostrobin did not possess antiestrogenic activity but presented anti-aromatase activity and decreased the growth of MCF-7 cells induced by DHEAS and E(2). This study provides particularly evidence of the potential biological interest of pinostrobin among the flavonoids. PMID- 10840158 TI - CYP17 polymorphism in the groups of distinct breast cancer susceptibility: comparison of patients with the bilateral disease vs. monolateral breast cancer patients vs. middle-aged female controls vs. elderly tumor-free women. AB - The CYP17 gene encodes an enzyme involved in several critical steps of steroidogenesis. The promoter region of the CYP17 displays a single-nucleotide polymorphism, which is suspected to modulate the expression of the gene and thus may contribute in the interindividual variations of hormonal background. In agreement with this functional hypothesis, the MspA1+ allele (designated as A2) of the CYP17 was shown to render an increased risk of breast cancer (BC). However, the latter observation was disputed by a series of negative reports. Here, we re-evaluated the role of CYP17 MspA1 polymorphism in the BC susceptibility, using a non-traditional design of a case-control study. In addition to randomly selected 183 BC patients and 107 female middle-aged donors, we examined the groups with apparently extreme characteristics of either BC risk or BC resistance, namely the 57 bilateral breast cancer (biBC) patients and 75 elderly (>/=75 years old) tumor-free women. Neither BC nor biBC patients showed increased prevalence of 'unfavorable' A2 allele as compared with the non-affected cohorts. Moreover, the A2 variant was not significantly associated with the tumor size, nodal involvement and menopausal status in the patients either with the monolateral or bilateral disease. Thus, our data argue against the earlier reported role of the CYP17 in BC predisposition and progression. In addition, usual distribution of the CYP17 alleles in the elderly group indicates a neutral effect of this polymorphism on the longevity in females. PMID- 10840159 TI - Formation of abasic sites in DNA by t-butyl peroxyl radicals: implication for potent genotoxicity of lipid peroxyl radicals. AB - We investigated abasic site formation in calf thymus DNA after exposure to a model compound of lipid-derived peroxyl radical that was generated by the reaction of tert-butyl hydroperoxide (t-BuOOH) with hemoglobin. Abasic site density in DNA was quantified by use of an enzyme-linked immunosorvent assay-like assay. In the presence of 10 mM t-BuOOH and 12.5 or 25 microM hemoglobin, 0.6-1.0 abasic sites/10(4) nucleotides were formed. However, abasic sites were not detected after replacing hemoglobin with nonheme iron, e.g. EDTA/Fe(2+), which initiates the production of alkyl and alkoxyl radicals. Therefore, the present results suggest that lipid peroxyl radicals may have a genotoxic potential through unique reactions, including depurination and depyrimidination, which lead to DNA strand breakage. PMID- 10840160 TI - Inhibitory effect of a prostaglandin E receptor subtype EP(1) selective antagonist, ONO-8713, on development of azoxymethane-induced aberrant crypt foci in mice. AB - We previously reported that prostaglandin E(2) contributes to colon carcinogenesis through its binding to the prostaglandin E receptor subtype EP(1) using a genetic approach in EP(1)-knockout mice and a pharmacological approach with the EP(1) selective antagonist, ONO-8711. In the present study, we examined the effects of another more selective EP(1) receptor antagonist, ONO-8713, on development of azoxymethane (AOM)-induced aberrant crypt foci (ACFs) in male C57BL/6J mice treated i.p. with 10mg/kg body weight AOM once a week for 3weeks. Administration of ONO-8713 at doses of 250, 500 and 1000ppm in diet during and post-AOM treatment for 5weeks resulted in a dose-dependent reduction of ACF formation, being 15, 30 and 36% inhibition of the control value, respectively. The level of inhibition was similar to that with ONO-8711. Moreover, ONO-8713 suppressed the development of ACF when administered at post-AOM, as in the case of ONO-8711. The data confirm EP(1) receptor involvement in colon carcinogenesis. PMID- 10840161 TI - p53-dependent expression of PIG3 during proliferation, genotoxic stress, and reversible growth arrest. AB - The p53-inducible gene 3 (PIG3) was recently identified in a screen for genes induced by p53 before the onset of apoptosis. PIG3 shares significant homology with oxidoreductases from several species. In this study, PIG3-specific antibodies were used to analyze cellular PIG3 protein levels under control and genotoxic stress conditions. PIG3 protein was localized to the cytoplasm and induced in primary, non-transformed, and transformed cell cultures after exposure to genotoxic agents. The induction of PIG3 was p53-dependent and occurred with delayed kinetics as compared with other p53 downstream targets, such as p21 and MDM2. Using a p53-inducible cell model system, in which p53-mediated growth arrest is reversible, we found that PIG3 levels were increased during p53 mediated growth arrest. Interestingly, elevated levels of PIG3 were maintained in cells that resumed cycling in the absence of ectopic p53 expression, suggesting that PIG3 is a long-lived reporter, which may be useful for detecting transient activation of p53. PMID- 10840162 TI - Association between GSTM1*0 and squamous dysplasia of the esophagus in the high risk region of Linxian, China. AB - Individuals with specific phase I and phase II enzyme polymorphisms may be at increased risk for squamous cell carcinoma of the esophagus. However, to our knowledge there has been only one previous report that evaluates a potential role for these polymorphisms in increasing risk for preneoplastic squamous lesions of the esophagus. To explore this further, we examined polymorphisms in CYP1A1, CYP2E1, GSTM1 and GSTT1, both independently and in combination, for potential associations with the risk of biopsy-proven squamous dysplasia of the esophagus in asymptomatic adults from Linxian, a high risk region in China. Cases consisted of 56 individuals from an esophageal cancer screening study with an endoscopic biopsy diagnosis of mild or moderate squamous dysplasia. Each case was matched on age (+/- 1 year) and gender to a control. Controls were defined as screening study participants with an endoscopic biopsy diagnosis of normal mucosa or esophagitis. DNA was extracted from frozen cell samples obtained by cytologic balloon examination and genotyped using standard methods. Individuals who were GSTM1 null (homozygous for GSTM1*0) were found to have a tendency for an increased risk of esophageal squamous dysplasia (odds ratio=2.6, 95% CI, 0.9 7.4). No excess risks were observed for inheritance of other putative at risk genotypes CYP1A1*2B, CYP2E1*6 or GSTT1*0. The risk associated with the inheritance of combined genotypes was not significantly different than the risk estimates from the univariate analysis. These results are consistent with the notion that exposure to environmental carcinogens that are detoxified by GSTM1, such as polycyclic aromatic hydrocarbons, may contribute to the etiology of esophageal cancer in Linxian. PMID- 10840163 TI - Granulocyte-macrophage colony-stimulating factor upregulates matrix metalloproteinase-2 (MMP-2) and membrane type-1 MMP (MT1-MMP) in human head and neck cancer cells. AB - Matrix metalloproteinase-2 (MMP-2) and membrane type 1-MMP (MT1-MMP) play an important role in the invasion and metastasis of head and neck squamous cell carcinoma (HNSCC), but the mechanism of their regulation is not clearly understood. Recently, granulocyte-macrophage colony-stimulating factor (GM-CSF) has been shown to be associated with cancer invasion and metastasis. We hypothesized that GM-CSF may upregulate MMP-2 and/or MT1-MMP expression in HNSCC cells, and may thereby influence their ability to invade and metastasize. We studied the effects of GM-CSF on the production of MMP-2 and MT1-MMP in HNSCC cell lines SAS and HSC-2. Gelatin zymography of conditioned media derived from HNSCC cells revealed a major band of 68 kDa, which was characterized as proMMP-2. GM-CSF stimulated the production of proMMP-2 in both cell lines in a dose dependent manner. Treatment with 50 ng/ml GM-CSF for 24 h increased the proMMP-2 activity 3.4-fold in SAS cells and 2.3-fold in HSC-2 cells compared with untreated controls. Northern blot analyses demonstrated that GM-CSF led to elevated mRNA levels of MMP-2 and MT1-MMP in both cell lines. The results identify GM-CSF as a regulator of MMP-2 and MT1-MMP expression in certain types of HNSCC, and suggest that GM-CSF may contribute to the invasiveness of HNSCC through the regulation of MMP-2 and MT1-MMP expression. PMID- 10840164 TI - Glutathione S-transferase-pi gene expression and platinum drug exposure in human lung cancer. AB - We examined the association between the gene expression levels of glutathione S transferase-pi (GST-pi) and platinum drug exposure in human lung cancer. First we monitored GST-pi gene expression levels in two lung cancer cell lines and in peripheral mononuclear cells of ten previously untreated lung cancer patients after platinum drug exposure. Next we examined GST-pi gene expression levels in 40 lung cancer autopsy specimens. The GST-pi gene expression levels were assessed by the quantitative reverse transcription-polymerase chain reaction or Northern blot analysis. The GST-pi gene expression was not induced by platinum drugs either in vitro and in vivo within 24 h of exposure. In contrast, GST-pi gene expression levels in lung cancer tissues of patients who had been exposed to platinum drugs at least 1 month before death were significantly higher than that in those of patients who had not been exposed. These results suggest that GST-pi gene expression is associated with chronic exposure to platinum drugs in lung cancer and/or the stress response to xenobiotics. PMID- 10840165 TI - Induction of the angiogenic factor VEGF in the uterus by the antiprogestin onapristone. AB - Onapristone (also referred to as ZK 98,299) is an antiprogestin that shares a number of structural similarities to mifepristone (RU-486) and other drugs in this class. While investigating the actions of antiprogestins on steroid hormone induced gene expression of angiogenic factors such as vascular endothelial growth factor (VEGF), we noted that onapristone alone induces VEGF transcript levels in the immature, ovariectomized rat uterus. In addition, onapristone induces expression of c-fos mRNA, which is induced by estrogens but not progestins in this target tissue. This induction of VEGF and c-fos by onapristone is inhibited by the antiestrogen ICI 182,780, but not by the antiprogestin RU-486. Both transcripts are very rapidly induced by onapristone, with maximal mRNA levels observed 3-6 h after in vivo administration of the drug. This time course is similar to that for induction of these genes by estrogenic hormones. Dose response studies show that both these genes are maximally induced by a 2.5 mg/kg dose of onapristone following intra peritoneal injection. These results indicate that onapristone rapidly upregulates several genes normally under estrogenic regulation in the immature rat uterus. Importantly, this is the first report of the induction of a major angiogenic factor by an antiprogestin. Since an increase in vascularity increases tumor expansion and metastasis, the induction of angiogenesis and its regulatory factors such VEGF may be an important end-point to consider in the development and use of antiprogestins for the treatment of neoplastic disease. PMID- 10840166 TI - Highest microvessel count as a long-term prognostic factor in Japanese breast cancer patients. AB - The aim of this study was to determine whether microvessel density (MVD) could add useful information in predicting the prognosis of breast cancer patients. In our study, MVD was calculated by counting microvessels per x200 field in the highest neovascularized area of the tumor (highest microvessel count, HMC). HMC significantly increased according to the increased number of positive nodes. Higher HMC significantly correlated with worse relapse-free survival (RFS) of patients with negative node, one to three positive nodes in the axilla or with stage I and II tumors. HMC, however, was not predictive for RFS of patients with four or more positive nodes or with stage III tumors. Multivariate analysis revealed that HMC was second only to nodal status and tumor size as being predictive for RFS. These results suggest that HMC could be used in selection of patients with early-stage breast cancer who are at high risk for having occult metastasis. PMID- 10840167 TI - Acute stress, memory, attention and cortisol. AB - An investigation was conducted to explore the relationship between acute changes in cortisol and memory and attention in the context of an acute naturalistic stressor, namely, examination stress. Sixty students (36 male, 24 female) participated in an assessment of self-reported levels of stress, salivary cortisol, short term memory, selective and divided attention and auditory verbal working memory. Assessments were conducted during a non-exam and exam period. The results revealed that the exam period was associated with an increase in perceived levels of stress, but also a significant reduction in levels of salivary cortisol, compared with the non-exam period. This reduction in cortisol was associated with enhanced short-term memory (as measured by the total number of words recalled in a free recall task), impaired attention and an impairment in the primacy effect (a hippocampal-specific index of short term memory), but no significant effects on auditory verbal working memory. It was concluded that the results support the view that cortisol can modulate cognitive processes and that the effects of corticosteroids on cognitive function are selective. PMID- 10840168 TI - Beta-endorphin (1-31) in the plasma of male volunteers undergoing physical exercise. AB - beta-Endorphin is an opioid peptide representing the C-terminal 31 amino acid residue fragment of proopiomelanocortin (POMC). The release of beta-endorphin from the pituitary into the cardiovascular compartment under physical or emotional stress has been frequently reported. However, besides beta-endorphin (1 31), nine acetylated or non-acetylated beta-endorphin analogues exist - in addition to N-terminally elongated beta-endorphin derivatives such as beta lipotropin (beta-LPH). Since conventional radioimmunoassays (RIAs) and even commercially available two site-RIAs pick up at least some of those beta endorphin derivatives, only "beta-endorphin immunoreactive materials" and not authentic beta-endorphin have been determined in those studies. We have developed a highly specific two site-RIA for beta-endorphin (1-31), which does not cross react with all beta-endorphin derivatives known to occur as yet. Using this RIA as well as further assays for determination of beta-endorphin (1-31), beta endorphin immunoreactive material (IRM), ACTH and Cortisol in the plasma of 14 volunteers upon intensive physical exercise, we found authentic beta-endorphin only in about 50% of the plasma samples, representing therein only a minor portion of the beta-endorphin IRM. PMID- 10840169 TI - Effects of testosterone administration on selective aspects of object-location memory in healthy young women. AB - Previous work has indicated that object-location memory is sensitive to sex differences as well as variations in the menstrual cycle. The goal of the present study was to further examine the hormonal basis of human spatial memory by assessing the effects of a single dose of exogenous testosterone in healthy young women on three recall conditions: positional reconstruction; object-to-position assignment; and the combined condition in which subjects both have to reconstruct the precise locations and to link the different objects to the correct places. In the latter condition, delayed recall (3 min delay) improved with testosterone. Although the effects were only small and need further substantiation, they support the idea that testosterone may have an activational effect on selective aspects of cognitive functioning. PMID- 10840170 TI - Conscious and preconscious selective attention to social threat: different neuroendocrine response patterns. AB - This study was designed to investigate the relationship between selective attention to social threat and neuroendocrine activity. Selective attention to social threat was measured using a supraliminal (unmasked) and a subliminal (masked) version of a pictorial emotional Stroop task, comparing color-naming latencies of neutral and angry faces. Neuroendocrine activity was assessed as (pre-task to post-task) increases in salivary cortisol and testosterone. Forty subjects were randomly assigned to the unmasked or masked version of the task. Analyses for the unmasked task revealed that post-task cortisol levels were significantly increased in subjects showing selective attention to angry faces. Results for the masked task indicated that post-task cortisol and testosterone levels were significantly increased in subjects showing preconscious selective attention to angry faces. The difference in neuroendocrine activity between tasks is suggested to depend on cortical (i.e. prefrontal) control in the unmasked task. Thus, psychological affective regulatory processes were involved in the unmasked task, whereas the neuroendocrine response patterns in the masked task indicates a biologically prepared mechanism. PMID- 10840171 TI - Elevated sorbitol concentration in the cerebrospinal fluid of patients with mood disorders. AB - This study was undertaken to test the hypothesis that a specific pathophysiological mechanism of diabetic neuropathy, namely increased polyol pathway flux, could be operative in patients with bipolar and unipolar mood disorders. Numerous studies have shown abnormalities of carbohydrate metabolism, including high rates of diabetes mellitus, in patients with mood disorders. Several studies have found that peripheral neuropathy is a risk factor for depression in diabetics. Furthermore, increased polyol pathway flux results in elevated sorbitol concentrations in peripheral tissues and cerebrospinal fluid (CSF) of diabetics with neuropathy. The purpose of this study was to determine whether sorbitol concentration is elevated in the CSF of non-medically ill patients with mood disorders. Lumbar punctures were performed on 30 subjects - 10 with bipolar mood disorder, 10 with unipolar mood disorder, and 10 age-matched normal controls, and CSF sorbitol concentrations were measured, using a gas chromatographic-mass spectroscopic technique. The mean+/-standard deviation of CSF sorbitol concentrations differed among the three groups as follows: bipolar (22.9+/-4.6 micromoles/l) > unipolar (19.0+/-2.8 micromoles/l)>normal control (15. 6+/-1.9 micromoles/l). One-way ANOVA showed significant (P=0.0002) differences among the three groups. Post-hoc tests indicated a significant (P<0.05) difference between bipolars and normal controls, bipolars and unipolars, and unipolars and normal controls. Further investigation is needed to determine the pathophysiological significance of this novel finding of elevated sorbitol concentration in the CSF of patients with mood disorders. PMID- 10840172 TI - Low dose zolmitriptan as a 5-HT neuroendocrine challenge agent in humans. AB - The 5-HT1B/D agonist sumatriptan has been used in a number of studies as a neuroendocrine challenge agent. Whether its neuroendocrine effects are centrally mediated is unclear, however, since sumatriptan shows minimal penetration of the central nervous system. Zolmitriptan shows a greater penetration into the central nervous system than sumatriptan, and has recently been shown to be an effective challenge agent. In order to determine the neuroendocrine, temperature and side effects of a 2.5 mg oral dose of zolmitriptan, 17 healthy volunteers underwent a placebo controlled, repeated measures, double blind neuroendocrine challenge. Zolmitriptan or placebo were administered, and cortisol, growth hormone, prolactin, blood pressure and temperature, were measured over four hours after the dose of zolmitriptan. Zolmitriptan at this dose was well tolerated by all subjects, with minimal side effects and only minor effects on blood pressure. There was a significant increase in serum growth hormone after zolmitriptan compared to placebo, however there were no significant effects on cortisol, prolactin or oral temperature. The neuroendocrine effects of 2.5 mg of orally administered zolmitriptan are similar to previously reported effects of sumatriptan, with minimal side effects. PMID- 10840173 TI - Consequences of the presence of the mother or unfamiliar adult female on cortisol, ACTH, testosterone and behavioral responses of periadolescent guinea pigs during exposure to novelty. AB - Periadolescent guinea pigs were exposed to a novel environment for 10, 30, 60, or 90 min while either alone or with either their biological mother or an unfamiliar adult female. Both classes of females reduced plasma cortisol, ACTH and vocalization responses to the novel environment, and did so to an equivalent degree. However, behavioral interactions with the mother and unfamiliar female differed considerably. The offspring exhibited more defensive behavior and were the targets of more agonistic acts when with the unfamiliar female, though one agonistic behavior, kicking, was observed more often in tests with the mother. Males displayed more social/courtship behavior and tumescence with the unfamiliar female. Further, males exhibited higher plasma testosterone levels when with the unfamiliar female than when either with the mother or alone. These data in conjunction with earlier findings suggest that the ability of unfamiliar adult females to moderate hypothalamic-pituitary-adrenal activity fully emerges during the periadolescent period, is not dependent on nurturant behavior by the adult female, and may facilitate the redirection of social behavior from the mother to unrelated adults. Further, mothers appear to inhibit maternally directed sexual behavior and plasma testosterone elevations in their periadolescent sons, effects which likely serve to inhibit inbreeding. PMID- 10840174 TI - Neurohormonal responses to cholecystokinin tetrapeptide: a comparison of younger and older healthy subjects. AB - We recently found that, compared with younger healthy subjects, older healthy subjects had less symptomatic and cardiovascular response to the panicogenic agent cholecystokinin tetrapeptide (CCK-4). As an exploratory part of that study, we also evaluated the effect of aging on neurohormonal responses to CCK-4. These hormonal data are the focus of this article. Forty healthy volunteers aged 20-35 years and 40 healthy volunteers aged 65-81 years, divided equally between men and women, were compared on their hormonal responses (maximum change from baseline in growth hormone [GH], prolactin, adrenocorticotropic hormone [ACTH], and cortisol) to the intravenous administration of 50 microg of CCK-4 or placebo. Blood samples for serum hormone determination were collected at 2 minutes prior to the intravenous challenge (baseline) and at 2, 5, and 10 minutes after the challenge. In both age groups, maximum increase in prolactin, ACTH and cortisol was significantly greater with CCK-4 than with placebo. Following administration of CCK-4, younger and older groups did not significantly differ in maximum increase in prolactin, ACTH, or cortisol. Older subjects had a statistically significant smaller increase in GH compared with younger subjects but the magnitude of the difference was small and of doubtful clinical relevance. Older subjects who had a panic attack had significantly greater elevations of all hormones compared with those who did not panic and younger panickers had a significantly greater elevation of GH compared with young nonpanickers. For the most part, maximum changes in hormonal levels were not correlated with symptom severity, suggesting that other factors may have contributed to the differential effect of panic on the HPA axis. PMID- 10840175 TI - Prenatal exposure of rats to valproic acid reproduces the cerebellar anomalies associated with autism. AB - Abnormalities in anatomy and function of the cranial nerve motor nuclei have been demonstrated in some people with autism and can be modeled in rats by exposure to valproic acid during neural tube closure. Reductions in Purkinje cell number and cerebellar volume, particularly of the posterior lobe, have also been reported in people with autism. Thus, a stereological examination of cerebellar morphology was undertaken in valproate-exposed rats. Compared to controls, rats exposed to a single dose of 600-mg/kg sodium valproate on embryonic day 12.5 had significantly fewer Purkinje cells in the cerebellar vermis and a reduction short of significant in the hemispheres. The diminished cell numbers reflect reductions in tissue volume throughout the cerebellum, rather than cell density, which was unaffected in all regions. Within the vermis, the reduction in volume was significantly greater in the posterior lobe than in the anterior lobe. The results parallel those reported for human cases of autism. PMID- 10840176 TI - Effects of prenatal marijuana exposure on child behavior problems at age 10. AB - This is a prospective study of the effects of prenatal marijuana exposure on child behavior problems at age 10. The sample consisted of low-income women attending a prenatal clinic. Half of the women were African-American and half were Caucasian. The majority of the women decreased their use of marijuana during pregnancy. The assessments of child behavior problems included the Child Behavior Checklist (CBCL), Teacher's Report Form (TRF), and the Swanson, Noland, and Pelham (SNAP) checklist. Multiple and logistic regressions were employed to analyze the relations between marijuana use and behavior problems of the children, while controlling for the effects of other extraneous variables. Prenatal marijuana use was significantly related to increased hyperactivity, impulsivity, and inattention symptoms as measured by the SNAP, increased delinquency as measured by the CBCL, and increased delinquency and externalizing problems as measured by the TRF. The pathway between prenatal marijuana exposure and delinquency was mediated by the effects of marijuana exposure on inattention symptoms. These findings indicate that prenatal marijuana exposure has an effect on child behavior problems at age 10. PMID- 10840177 TI - Effects of chronic lead exposure on learning and reaction time in a visual discrimination task. AB - Long-Evans rats exposed chronically to lead (Pb) acetate (0, 75, or 300 ppm) were tested as adults on an automated, three-choice visual discrimination task as part of a larger study designed to elucidate the cognitive effects of developmental Pb exposure. Median adult BPb levels for the groups were <5, 20, and 36 microgram/dl. The pattern of results suggested a linear effect, with increasing lead dose producing progressively slower learning and an increased incidence of "impaired" individuals. This latter measure proved to be slightly more sensitive than the former, suggesting individual differences in susceptibility to Pb neurotoxicity. Additional analyses revealed that the impairing effect of Pb was seen in both the chance and post-chance learning phases, indicating that the deficit was not limited to (but could include) attentional function. Reaction time on incorrect trials was reduced in the 300-ppm group, whereas no Pb effect was seen for correct trials. The present findings suggest that chronic developmental Pb exposure produces an associative deficit as well as a tendency to respond rapidly, but does not affect information-processing speed. PMID- 10840178 TI - Lead absorption and psychological function in Zagreb (Croatia) school children. AB - A cross-sectional study was performed on 275 pupils from the third and fourth grade of three elementary schools (three urban areas with different traffic conditions) in Zagreb. Lead exposure was environmental, mostly through leaded gasoline. The difference in traffic density around the schools was consistent with biological indicators of lead absorption. The aim of the study was to clarify the relationship between characteristic biological indicators of lead absorption including indicators of hematological status with some psychological functions. Lead absorption in pupils was relatively low (mean blood lead: 70.8 +/ 17.88 microgram/L). Pupils' socio-economic status was evaluated by parents' education. The results obtained indicate that gender and school were associated with both biological and psychological variables. After adjusting for age, parental education, and gender, lead appears to have no association with cognitive or psycho-motor measures. The nonstandardized regression coefficients for blood lead-as a measure of the size of lead effect on VIQ, NIQ, and IQ-were 0.016, -0.031, and -0.025, respectively, all nonsignificant. PMID- 10840179 TI - The influence of route of administration on the acute cardiovascular effects of cocaine in conscious unrestrained pregnant rats. AB - The intravenous route of administration, accessed via a subcutaneous vascular access port, has been recently suggested as an animal model for studying the developmental effects of maternal cocaine abuse in the pregnant and/or group housed rat. The present study (1) assessed the cardiovascular effects of intravenous (IV) cocaine, delivered via bolus injection, in chronically catheterized near-term pregnant rats, and (2) compared the IV cardiovascular responses to those following cocaine delivered via the commonly employed subcutaneous (SC) and intragastric (IG) routes of administration. Pregnant gestation day 15 (GD15) young adult female Sprague-Dawley rats (n = 21) were anesthetized and catheters surgically implanted into the carotid artery, jugular vein, fundus of the stomach, and a subcutaneous pouch. On GD17-19, heart rate (HR) and mean arterial pressure (MAP) were assessed, using a within-subjects design, prior and subsequent to IV (3 mg/kg), IG (60 mg/kg), and SC (40 mg/kg) cocaine. An interval of 6 h separated IV and IG cocaine administration and an interval of 18 h separated IG and SC cocaine administration. The peak responses of HR (23% downward arrow) and MAP (37% upward arrow) following IV cocaine were noted within 0.5 min. In contrast, the peak responses of HR (4% downward arrow, 6% downward arrow) and MAP (2% upward arrow, 15% downward arrow) after IG (23 min) or SC (26 min) cocaine, respectively, were significantly smaller and markedly delayed. No significant change in aortic blood flow velocity was detected following cocaine via any route of administration, although phasic flow velocities (PFV) were differentially sensitive to route of administration (PFV(dias) not PFV(sys)); IV cocaine increased (55% upward arrow) whereas IG or SC cocaine decreased approximately 35% downward arrow) PFV(dias). The pressor effects of an equimolar dose of IV cocaine methiodide (3.9 mg/kg) were indistinguishable from those of IV cocaine (38% upward arrow vs. 37% upward arrow), as were the effects on PFV(dias) (83% upward arrow vs. 55% upward arrow). The lack of an effect of cocaine methiodide on HR was consistent with the bradycardia effect of cocaine attributable to central mediation of the baroreflex. Finally, the pressor effects of IV cocaine paralleled the rapidly peaking arterial plasma levels of cocaine noted within 30 s after the initiation of drug injection. In sum, prominent effects of IV cocaine on maternal cardiovascular physiology are noted; as such, the recent reports of a lack of maternal/fetal toxicity following daily (3-6mg/kg) IV cocaine during GD8-21 are not due to use of an ineffective drug dose. It was equally clear that the SC and IG routes of exposure did not reproduce the cardiovascular component(s) of the expected physiological response to cocaine. PMID- 10840180 TI - Prenatal exposure to anti-HIV drugs. long-term neurobehavioral effects of lamivudine (3TC) in CD-1 mice. AB - The present study was aimed at investigating the long-term effects of prenatal exposure to lamivudine (3TC), an antiretroviral drug used in clinical practice alone or in combination with zidovudine (AZT) to prevent mother-to-child transmission of the HIV virus. Pregnant CD-1 mice were given per os twice daily either 3TC at different doses (125, 250, or 500 mg/kg) or vehicle solution (NaCl 0. 9%) from pregnancy day 10 to delivery. Offspring behavior was examined on postnatal day 35 in a 20-min social interaction test. At adulthood different behavioral endpoints were analyzed, including locomotor activity and exploration in an open field following administration of the muscarinic antagonist scopolamine (2 mg/kg), spatial learning in either radial arm or Morris water maze, virgin female behavior in a maternal induction test, and pain sensitivity in a hot-plate test (52 +/- 0.1 degrees C). Our findings confirm the low neurotoxicity of 3TC in comparison to AZT. However some significant behavioral alterations were found, namely (1) a decrease in immobility in the open field test, (2) an increase in the responsiveness to scopolamine shown by the 500-mg/kg 3TC mice (sniffing behavior) in the open field, and (3) a longer escape latency in the first day of the reversal phase in the Morris task (particularly marked in the 250-mg/kg treatment group). No significant changes in either pain sensitivity, social/affiliative, or maternal behavior were found, although a higher occurrence of aggressive behavior toward foster pups was noted in both 125 and 500-mg/kg 3TC females. PMID- 10840181 TI - Age-related impairment of long-term depression in area CA1 and dentate gyrus of rat hippocampus following developmental lead exposure in vitro. AB - Chronic developmental lead exposure is known to be associated with cognitive dysfunction in children. Impairment of the induction of long-term depression (LTD) has been reported in area CA1 and dentate gyrus (DG) of rat hippocampus following chronic lead exposure. The present study was carried out to investigate age-related alterations of LTD in area CA1 and DG of rat hippocampus following developmental lead exposure in vitro. Neonatal Wistar rats were exposed to lead from parturition to weaning via milk of dams drinking 0.2% lead acetate solution. Field excitatory postsynaptic potentials (EPSPs) were recorded in hippocampal slices at various postnatal ages: postnatal day (PND) 17-23, 27-33, and 57-63. Following low-frequency stimulation (LFS, 900 pulses/1 Hz), the average magnitude of LTD is age related. In the controls, LTD magnitude in area CA1 decreased with age, whereas in DG it increased with age. In the lead-exposed groups, the magnitude of LTD declined during development in both area CA1 and DG. The differences of LTD magnitude between the control and lead-exposed rats were 27.26 +/- 9.15% (PND 17-23), 21.59 +/- 12.93% (PND 27-33), and 16.96 +/- 9.33% (PND 57 63) in area CA1, and were 6.95 +/- 9.26%, 17.60 +/- 3.91%, and 33.63 +/- 10.47% in DG, respectively. These results demonstrated that the lead-induced impairment of LTD magnitude was an age-related decline in area CA1 and an age-related increase in area DG of rat hippocampus. Published by Elsevier Science Inc. PMID- 10840182 TI - Effects of cholinesterase inhibitors on a two-component chained schedule performance in rats. AB - The acetylcholinesterase (AChE) inhibitors physostigmine (PHY), tacrine (THA), and heptylphysostigmine (HEP) have been evaluated as potential therapeutics for treatment of Alzheimer's disease (AD) and as prophylactics against organophosphate (OP) poisoning. The above medical applications are based upon the neurochemical principles of elevation of transient levels of acetylcholine (ACh) in brain and reversible inhibition of AChE in blood and brain, respectively. The present study was undertaken to evaluate the effects of these drugs on performance of a two-component chained schedule of differential-reinforcement-of high-rate (DRH) reward/differential-reinforcement-of-low-rate (DRL) nonreward contingencies, for water reinforcement in 2-h experimental sessions in rats. Both PHY (0.031-0.25 mg/kg, SC) and HEP (0.625-10.0 mg/kg, SC) decreased overall reinforcement rate and nonreinforced response in a dose-related and parallel manner, whereas THA (0.625-5.0 mg/kg) decreased the overall reinforcement rate in a dose-related manner, but did not significantly affect nonreinforced response. The least significant doses of PHY (0.625 mg/kg), THA (1.25 mg/kg), and HEP (1. 25 mg/kg) on the behavioral performance were associated with oral movements and/or muscle fasciculation. Moderate to high doses of these drugs (i.e., PHY >/= 0.624, THA >/= 1.25, and HEP >/= 1.25 mg/kg) produced behavioral suppression, which resulted mostly from the cessation of responding in the presence of cholinergic adverse events. The ED(50) values of the behavioral disruption (as documented by overall reinforcer loss) for PHY, THA, and HEP were 0. 081, 3.87, and 2.89 mg/kg, respectively, and the behavioral-deficit-free (BDF) doses were 0.031, 0.625, and 0.625 mg/kg, respectively. Preclinical data revealed that the BDF doses of both PHY and HEP have moderate inhibition of AChE, which leads to an efficacious elevation of ACh level in the brain, whereas the BDF dose of THA shows no significant AChE inhibition or elevation of brain ACh level; however, they all have cognition enhancing effects at their respective safety doses. The above data suggest that the BDF doses of PHY and HEP may have prophylactic efficacy against OP poisoning, whereas THA may not. The BDF doses of these three drugs are comparable to the maximum tolerated doses in clinical practice, suggesting that the present rat model may have potential value in predicting the clinical safety of AChE inhibitors developed for therapy of AD and prophylaxis against OP poisoning as well. PMID- 10840183 TI - Comparison of neurobehavioral changes in three inbred strains of mice prenatally exposed to methylmercury. AB - Pregnant mice of three inbred strains (BALB/c, C57BL/6J, C57BL/6Cr) were orally given methylmercury (MMC; 3 x 3 mg/kg body weight) or the equivalent volume of phosphate-buffered saline during days 12-14 of gestation and allowed to deliver. The behaviors of their male offspring were evaluated in an open field and their home cage and in a Morris water maze. In the open field test, the BALB/c and C57BL/6Cr MMC groups exhibited less total locomotor activity than did their respective control groups. However, there was no significant difference observed between the MMC and control C57BL/6J strain. In the BALB/c strain, the MMC group exhibited significantly more central locomotion and significantly less peripheral locomotion than did the control group. These results indicated that the prenatal exposure to MMC caused decreases in open-field activity in the C57BL/6Cr and BALB/c strains, concomitantly with a change in emotional status in BALB/c strain. For spontaneous activity in their home cage, all groups moved more actively in the dark phase than in the light phase except BALB/c MMC group. The BALB/c MMC group moved in the light phase as much as in the dark phase, indicating a disturbance of nocturnal rhythm of spontaneous activity. In the Morris water maze, the C57BL/6Cr and C57BL/6J control groups perform very well over the 5 consecutive days. The prenatal exposure to MMC caused significantly prolonged latency in the C57BL/6Cr and C57BL/6J, but not in BALB/c strain. This result indicated that the prenatal exposure to MMC impaired the performance in the Morris water maze differently among the strains. This study provides a basis for evaluating strain-specific neurobehavioral changes when the widely used three inbred strains of mice are chronically exposed to MMC. PMID- 10840184 TI - Postnatal behavior in hatano high- and low-avoidance rats following prenatal exposure to low-dose methylazoxymethanol. AB - The hypothesis that genetic factors influence behavioral effects was tested in rats exposed prenatally to methylazoxymethanol (MAM). We examined whether baseline behavior is an important factor influencing behavioral effects, and whether a behaviorally selected strain was useful for study of neurobehavioral teratology. Pregnant high- and low-avoidance animals (HAAs and LAAs) of the Hatano strain, selectively bred for high and low shuttlebox avoidance responses, respectively, were given an IP injection of a low dose of MAM (15 mg/kg) on day 14 of gestation. The offspring of these animals were subjected to behavioral tests for locomotor activity (running-wheel and open-field tests) and learning ability (Biel maze and shuttlebox avoidance tests). There were no significant effects of MAM on running-wheel activity or shuttlebox avoidance learning, whereas the number of errors in the Biel maze was increased in the MAM offspring of both strains. Interestingly, open-field activity of the MAM offspring was markedly decreased in LAAs but not in HAAs. Therefore, an additional experiment was performed to determine plasma levels of ACTH and corticosterone following open-field exposure. When compared to control offspring of the respective strains, plasma levels of ACTH and corticosterone were not altered by prenatal MAM treatment in LAAs. Instead, the MAM offspring in HAAs exhibited decreased ACTH levels in absence of behavioral alterations. These results demonstrated that prenatal exposure to low doses of MAM may alter postnatal behavior and endocrine response of the offspring, although to a differing degree in HAAs and LAAs. Our observations suggested that behaviorally selected strains are sensitive to neurobehavioral teratogens such as MAM. PMID- 10840185 TI - Minimal behavioral effects from developmental cerebellar stunting in young rats induced by postnatal treatment with alpha-difluoromethylornithine. AB - Postnatal treatment with alpha-difluoromethylornithine (DFMO), a potent inhibitor of ornithine decarboxylase, reduces polyamine levels in rats. Because polyamines are critically involved in growth and development, body and/or brain weights are often decreased by DFMO treatment. Here, rats were injected subcutaneously with 0, 250, 500, or 750 mg/kg of DFMO on postnatal days (PNDs) 5-10. Behavioral assessments included righting reflex, negative geotaxis, forelimb hanging, open field activity, and rotarod performance. Additionally, day of eye opening was recorded and on PND 28, whole and regional brain weights were measured. Cerebellar/whole-brain ratio was decreased in a dose-dependent manner whereas frontal cortex/whole-brain ratio was increased. Eye opening was delayed to a similar extent in all treated groups whereas body weight was unaffected. alpha difluoromethylornithine treatment had no significant effects on the assessed behaviors. These results indicate that 6 days of DFMO treatment can substantially impact cerebellar development, but this appears to have few effects on these early assessed behaviors. However, potential behavioral alterations may not be apparent until adulthood. Published by Elsevier Science Inc. PMID- 10840186 TI - Effects of age and gender but not prenatal cocaine on random ratio and delayed spatial alternation responding in rats. AB - This investigation employed a longitudinal analysis of rat operant behavior under two different schedules of reinforcement following prenatal exposure to cocaine. Offspring were derived from four maternal exposure groups: 50 mg/kg cocaine, their pair-fed controls, 25 mg/kg cocaine, and freely fed controls. Cocaine was administered via gavage from gestation day 6-20. A maternal fostering procedure was used. Pairs of male and female littermates were assigned to a 7-, 14-, or 21 month cohort and at the appropriate age were trained to respond on one lever in a two-lever operant chamber. Reinforcement was delivered with a series of random ratio (RR) schedules where the RR value was increased across sessions. After RR training, animals were examined with a delayed spatial alternation (DSA) procedure in the same chambers. Male offspring responded at higher rates than females during high-probability RR schedules, whereas advancing age was associated with lower response rates during low-probability RR schedules in both males and females. Prenatal cocaine exposure exerted only limited effects on RR responding during transition and did not affect DSA behavior. The results of this longitudinal analysis suggest that prenatal cocaine does not exert global or far reaching learning deficits in prenatally exposed rats. PMID- 10840187 TI - Temporal development of 2',3'-dideoxyinosine (ddI)-induced peripheral myelinopathy. AB - The anti-HIV therapeutic dideoxyinosine (ddI) has been reported to produce a painful, dose-limiting peripheral myelinopathy in HIV-infected patients after chronic administration. We have previously demonstrated ddI-induced myelinopathy in a non-HIV-infected rat model after 20 weeks of dosing, characterized by myelin splitting and intramyelin edema. The present study examined the time course needed to produce the ddI-induced neuropathy. Adult male Sprague-Dawley rats were gavaged with vehicle or 415 mg/kg ddI twice daily for up to 20 weeks. Groups of treated (n = 6-8) and control (n = 3-5) animals were killed after 5, 10, 15, and 20 weeks of dosing and the distal end of the sciatic nerve was removed. The nerve was postfixed by immersion in neutral phosphate-buffered formalin, dehydrated in graded alcohols, and embedded in plastic embedding media. One-micrometer-thick sections were cut and stained with toluidine blue and basic fuchsin. Plasma levels of ddI on the day the animals were killed were greater than 10 microgram/ml within the first hour after dosing and fell rapidly to less than 1 microgram/ml (clinical range 1-2 microgram/ml) within 3 h after dosing. The abnormalities observed in the sciatic nerve were few, if any, after 5 or 10 weeks, but very prominent after 15 weeks of dosing. Four of the six ddI-treated rats exhibited abnormal morphology as evidenced by myelin splitting and ballooned myelin sheaths. Although abnormal morphology was present at 20 weeks of dosing, the effect was not as robust as at 15 weeks. This suggests that the nerve may partially recover from the effects of ddI with time. Published by Elsevier Science Inc. PMID- 10840188 TI - Editorial PMID- 10840189 TI - Modern bioavailability, bioequivalence and biopharmaceutics classification system. New scientific approaches to international regulatory standards. AB - In the last decade, the regulatory bioequivalence (BE) requirements of drug products have undergone major changes. The introduction of the biopharmaceutics drug classification system (BCS) into the guidelines of the Food and Drug Administration (FDA) is a major step forward to classify the biopharmaceutical properties of drugs and drug products. Based on mechanistic approaches to the drug absorption and dissolution processes, the BCS enables the regulatory bodies to simplify and improve the drug approval process. The knowledge of the BCS characteristics of a drug in a formulation can also be utilized by the formulation scientist to develop a more optimized dosage form based on fundamental mechanistic, rather than empirical, information. This report gives a brief overview of the BCS and its implications. PMID- 10840190 TI - Modelling during drug development. AB - With the advancement of both biological and computer sciences, new drug development faces the challenge to integrate a huge amount of knowledge accumulated from the very early quantitative structure-activity relationship investigations of the candidate molecule to the large scale clinical trials in patients. Whereas pharmacokinetics and pharmacodynamics are fields in which modelling has long demonstrated its value, its potential in many other areas of drug development has recently been the object of intensive scientific activity. The present review places emphasis on these newer applications; it includes the opinion of many experts in often highly specialised areas such as in vitro to in vivo extrapolation, toxicokinetics, non-continuous response models, population approaches and computer assisted simulation of clinical trials. It is most probable that in the near future many of these areas of research will be the objects of intensive and interesting developments. This will undoubtedly lead to improve developmental strategies for new drugs as well as more individualised pharmacological strategies for patients. PMID- 10840191 TI - Hydrogels in pharmaceutical formulations. AB - The availability of large molecular weight protein- and peptide-based drugs due to the recent advances in the field of molecular biology has given us new ways to treat a number of diseases. Synthetic hydrogels offer a possibly effective and convenient way to administer these compounds. Hydrogels are hydrophilic, three dimensional networks, which are able to imbibe large amounts of water or biological fluids, and thus resemble, to a large extent, a biological tissue. They are insoluble due to the presence of chemical (tie-points, junctions) and/or physical crosslinks such as entanglements and crystallites. These materials can be synthesized to respond to a number of physiological stimuli present in the body, such as pH, ionic strength and temperature. The aim of this article is to present a concise review on the applications of hydrogels in the pharmaceutical field, hydrogel characterization and analysis of drug release from such devices. PMID- 10840192 TI - Improving drug solubility for oral delivery using solid dispersions. AB - The solubility behaviour of drugs remains one of the most challenging aspects in formulation development. With the advent of combinatorial chemistry and high throughput screening, the number of poorly water soluble compounds has dramatically increased. Although solid solutions have tremendous potential for improving drug solubility, 40 years of research have resulted in only a few marketed products using this approach. With the introduction of new manufacturing technologies such as hot melt extrusion, it should be possible to overcome problems in scale-up and for this reason solid solutions are enjoying a renaissance. This article begins with an overview of the historical background and definitions of the various systems including eutectic mixtures, solid dispersions and solid solutions. The remainder of the article is devoted to the production, the different carriers and the methods used for the characterization of solid dispersions. PMID- 10840193 TI - HPMA copolymer-anticancer drug conjugates: design, activity, and mechanism of action. AB - The design, synthesis and properties of N-(2-hydroxypropyl)methacrylamide (HPMA) copolymers as carriers of anticancer drugs are reviewed. Macromolecular therapeutics based on HPMA copolymers are biocompatible, preferentially accumulate in tumors, and possess a higher anticancer efficacy than low molecular weight drugs. Novel designs of HPMA copolymer carriers resulted in long circulating conjugates and gene and oligonucleotide delivery systems. HPMA copolymer based macromolecular therapeutics were active against numerous cancer models and are in clinical trials. The data obtained indicated that macromolecular therapeutics activated different signaling pathways and possessed a different mechanism of action than free drugs. This bodes well for the success of future research aimed at identification of new intracellular molecular targets as a basis for the design of the second generation of macromolecular therapeutics. PMID- 10840194 TI - Local delivery of antimicrobial agents for the treatment of periodontal diseases. AB - Periodontitis is an inflammatory disease of the supporting tissues of the teeth caused by groups of specific microorganisms. Aggressive forms of periodontitis can be localized or generalized. The concept that localized problem sites may be treated by local drug delivery appears attractive as the antimicrobial agent is delivered within periodontal pockets and the therapy is targeted on specific pathogenic microorganisms. Local delivery of antimicrobial agents using controlled release systems should be considered as adjunctive to mechanical debridement for the treatment of localized forms of periodontal destruction. This article reviews various types of delivery systems evaluated in practical periodontal therapy. Despite the large number of studies showing an enhanced effectiveness of local antibiotherapy, there are insufficient comparative data to support any of the local delivery system. PMID- 10840195 TI - Cellular delivery of antisense oligonucleotides. AB - Antisense oligonucleotides can be successfully employed to inhibit specifically gene expression. However, many oligonucleotide classes are polyanions and cannot passively transit the cell membrane. Thus, the use of naked oligonucleotides for antisense purposes poses some rather stringent challenges, and it is not a trivial task to appropriately interpret the data derived from experiments in which they have been used. Multiple methods have been developed to improve intracellular, and in particular, intranuclear oligonucleotide delivery, and in doing so, to maximize the performance of the antisense technologies that are currently available. This review discusses the use of cationic lipids, protein and peptide delivery agents, and several novel chemical and viral methods that have recently been explored as delivery vehicles, focussing not only on their strengths, but also on their limitations. PMID- 10840196 TI - Poly(ortho esters) - their development and some recent applications. AB - Poly(ortho esters) have been under development since the early 1970s and four families of such polymers have been described. Of most interest are poly(ortho ester) III and poly(ortho ester) IV. Poly(ortho ester) III is a semisolid material that has been shown to be highly biocompatible and is currently being investigated as an adjunct to glaucoma filtering surgery and other ocular applications. However, the polymerization is difficult to control and is not readily scaled up. Poly(ortho ester) IV can be easily prepared in a highly reproducible manner, is very stable provided moisture is rigorously excluded and has also been shown to be highly biocompatible. It is currently under development for a variety of applications, such as ocular delivery, protein release, post operative pain treatment and post-operative cancer treatment. PMID- 10840197 TI - Revisiting PLA/PLGA microspheres: an analysis of their potential in parenteral vaccination. AB - Poly(lactide) and poly(lactide-co-glycolide) microspheres have been studied for controlled antigen delivery and immune response enhancement for more than a decade. Early developments of such vaccines were basically technology-driven, stemming from the well-established biocompatibility of these polymers in concert with their innate properties to tailor rates of bioerosion and release. More recently, other features have become equally or even more appealing, such as the adjuvancy of such microspheres and their ability to elicit cellular effector responses, so-called cytotoxic T-cell responses, in addition to antibody responses observed already in the very early studies. In this review, we intended to revisit microsphere-based vaccines designed for the parenteral route and attempted to outline major developmental issues, as well as to analyze immunological fundamentals and data associated with antigen delivery by microspheres. PMID- 10840198 TI - Biodegradable nanoparticles for oral delivery of peptides: is there a role for polymers to affect mucosal uptake? AB - Numerous authors have demonstrated uptake of micro- and nanospheres, consisting of natural or synthetic polymeric materials from the gastrointestinal tract over the past two decades. The exploitation of particulate carrier systems for the delivery of peptides and other hydrophilic macromolecules via the oral route remains a challenging task due to morphological and physiological absorption barriers in the gastrointestinal tract. This review examines recent progress in the field of nanoparticle uptake from this site of administration. Since most studies have been performed with poly(styrene) particles of different sizes relatively little is known about both the effect of physicochemical particle properties critical for absorption after peroral application, and the mechanisms of gastrointestinal particle uptake. Apart from particle size, type and composition of the polymers used for micro- or nanoencapsulation are crucial for an uptake and transport across mucosal barriers. Factors such as particle surface charge and hydrophilic/hydrophobic balance of these polymeric materials have not been investigated systematically since adjustment of these particle properties is almost impossible without synthetic modification of the polymers. The current findings will be reviewed and compared to those obtained with nanoparticles consisting of a novel class of charged comb polyesters, poly(2-sulfobutyl-vinyl alcohol)-graft-poly(D,L-lactic-co-glycolic acid), SB-PVAL-g-PLGA, allowing adjustment of physicochemical nanoparticle properties with a single class of polymers. PMID- 10840199 TI - Solid lipid nanoparticles (SLN) for controlled drug delivery - a review of the state of the art. AB - Solid lipid nanoparticles (SLN) introduced in 1991 represent an alternative carrier system to traditional colloidal carriers, such as emulsions, liposomes and polymeric micro- and nanoparticles. SLN combine advantages of the traditional systems but avoid some of their major disadvantages. This paper reviews the present state of the art regarding production techniques for SLN, drug incorporation, loading capacity and drug release, especially focusing on drug release mechanisms. Relevant issues for the introduction of SLN to the pharmaceutical market, such as status of excipients, toxicity/tolerability aspects and sterilization and long-term stability including industrial large scale production are also discussed. The potential of SLN to be exploited for the different administration routes is highlighted. References of the most relevant literature published by various research groups around the world are provided. PMID- 10840201 TI - Reconstructive surgery for deep venous reflux: a report on 144 cases. AB - This retrospective study was conducted on 144 lower extremities (133 patients) with deep venous reflux treated with surgery to restore venous valvular function. Clinically, 51% of patients were in class C5-C6, and based on etiology, patients were equally divided into primary and secondary venous disease. Four surgical procedures were used: valvuloplasty (n=85), transposition (n=18), transplantation (n=32), or Psathakis' technique II (n=9). The procedure chosen was determined mainly by the feasibility of the technique in the above-mentioned preferred order. Thus, 76% of valvuloplasties were performed for primary venous insufficiency. A postoperative venography routinely performed soon after surgery demonstrated a large number of segmental thromboses (20.3%). Their number was statistically different in primary and secondary (PTS) venous disease, respectively 8.8 vs 32.3%. Clinical and hemodynamic results were evaluated (duration of follow-up: 12-168 months) based on etiology and type of procedure. A correlation was established between clinical result (venous ulcer) and efficacy of valvular reconstruction. The latter was satisfactory in valvuloplasties (P=0.005) but not in venous transfer (P=0.35). Overall results were better for primary venous insufficiency than in postthrombotic syndromes (P=0.03). PMID- 10840200 TI - Self-dispersing lipid formulations for improving oral absorption of lipophilic drugs. AB - The main purpose of this review is to provide a current and general overview of the existing self-dispersing formulations resulting from dilution into emulsions, microemulsions and surfactant dispersions. The systematic approach used and the presentation of the various physico-chemical and biopharmaceutical aspects should facilitate the comprehension of this interesting field and clarify the main considerations involved in designing and characterizing a specific self dispersing drug delivery system. Studies have shown that the self-emulsification process is specific to the nature of the oil/surfactant pair, surfactant concentration, oil/surfactant ratio and temperature at which self-emulsification occurs. It was suggested that the ease of emulsification could be associated with the ease by which water penetrates into the various liquid crystalline (LC) or gel phases formed on the surface of the droplet. Numerous bioavailability studies carried out in animals and humans, reviewed in the present study, suggest that hydrophobic drugs are better absorbed when administered in self-dispersing lipid formulations (SDLFs). Examples which illustrate the beneficial use of SDLFs for drug absorption enhancement are presented. This review outlines SDLFs as one of the most promising approaches to overcome the formulation difficulties of these hydrophobic/lipophilic drugs. PMID- 10840202 TI - Complications of operative treatment of injuries of peripheral arteries. AB - In 1991 and 1992, a total of 97 patients with 106 peripherial arterial injuries underwent surgery at the Institute for Cardiovascular Diseases, Clinical Centre of Serbia. Civilian injuries accounted for 53 (54.6%) patients (94.3% males, age range: 16-63 yr, mean: 35.2), and 44 patients had war injuries (93.2% males, age range: 19-61 yr, mean: 34.8). The injuries affected the superfitial femoral artery in 31 (29.24%); the popliteal artery in 28 (26.41%); the brachial artery in 17 (16.04%); the posterior tibial artery in 6 (5.66%); the axillary artery in 5 (4.72%); the anterior tibial artery in 5 (4.72%); the tibioperoneal trunk in 4 (3. 77%); the common femoral artery in 4 (3.77%); the external iliac artery in 2 (1.89%); the profound femoral artery in 2 (1.89%); the radial artery in 1 (0.94%); and ulnar artery in 1 (0.94%).A total of 98 reconstructive procedures were used to treat these patients. Graft interposition carried out in 50 (51.02%); by pass in 25 (25. 51%); end-to-end anastomosis in 9 (9.18%); suture in 8 (8.16%); ligation in 4 (4.08%); and patch-angioplasty in 2 (2.04%). Primary reconstruction of injured arteries was performed in 72.2% and secondary repair in 27.8% cases. Infection developed in 51 (52.57%) patients, and it was significantly (P<0.05) more common in the war injuries (70.45%) and in secondary repairs (88.89%). The presence of associated lesions (69.56%) was also correlated with a greater rate of infection. Amputation was necessary in 21 (21.65%) of our patients, and was significantly (P<0.05) more often performed after secondary (44.44%) than primary operations (12.86%) and in the presence of associated injuries (32.61%). PMID- 10840203 TI - Identification of dendritic cells in ePTFE grafts explanted from humans. AB - Following implantation different cell types interact with synthetic vascular prostheses resulting in a complex immuno-inflammatory response. Dendritic cells are responsible for activating the primary T-lymphocyte immune response in various pathological conditions by their role as antigen-presenters. This study aimed at examining whether dendritic cells accumulate within small diameter expanded polytetrafluoroethylene (ePTFE, Goretex(R)) grafts explanted from humans. Segments of expanded polytetrafluoroethylene were explanted from 11 patients (6 male, 5 female), aged between 60 and 83 years (mean 70.7 years) at secondary or revision bypass operation. The graft implant duration varied from 4 months to 12 years (mean 40.5 months). Dendritic cells were identified immunohistochemically using S-100 antibody, as well as by electron microscopy. Immunohistochemical examination showed that all 11 explanted synthetic grafts contained S-100(+) cells colonising both the synthetic matrix itself, and the adjacent perigraft tissue. Electron microscopic analysis confirmed the presence of cells with a characteristic dendritic cell morphology within the grafts. Dendritic cells which accumulated within synthetic grafts were found to co localise with T-lymphocytes. Based on these observations, we speculate that dendritic cells may be involved in the immuno-inflammatory responses following the implantation of synthetic vascular prostheses through their interaction with T-lymphocytes. PMID- 10840204 TI - Paraanastomotic aneurysms of the abdominal aorta: a 15-year experience review. AB - The aim of the study is a retrospective review of clinical presentation and management of paraanastomotic aneurysms of the abdominal aorta (PAAA) surgically treated in our Department. From January 1984 to December 1998, 2183 aortic prosthetic grafts were implanted. During the same period, 24 patients were treated for PAAA, 19 false and five true aneurysms. Symptoms were present in 10 patients. Surgical management included tube grafting interposition (14), aortobifemoral bypass (2), graft removal with extraanatomic bypass (2) and with in situ revascularization by arterial homograft (4). Nine patients died during operation or in the early postoperative period, six died during follow-up. Mortality in symptomatic patients was 70%, while in asymptomatic group was 14% (P=0.01). Rupture of false PAAA was very frequent (47% of cases). PAAA are infrequent complications of proximal aortic graft revascularization and tend to be asymptomatic until rupture occurs. The incidence of mortality is very different in asymptomatic versus symptomatic group; rupture is particularly frequent in false PAAA, which must soon undergo surgery when diagnosed. Since PAAA may develop at any time after surgery, their incidence increase in relationship with the length of postoperative interval: therefore, all patients submitted to abdominal graft revascularization need a lifetime surveillance program. PMID- 10840205 TI - Delayed treatment of isthmic aortic rupture. AB - Traumatic rupture of the thoracic aorta is a life-threatening injury with a high mortality, and is difficult to manage in polytraumatized patients. Between 1980 and 1998, 50 patients were admitted to our Department with acute traumatic aortic rupture (TAR). The site of lesion was usually isthmic (86% of patients). From 1980 to 1992, 21 patients (Group I) underwent emergency surgical repair; from 1992 to January 1998, 29 patients (Group II) underwent intensive medical treatment, except one who was haemodynamically unstable and died 8h after the trauma from a massive haemothorax before an emergency thoracotomy could be attempted. The aortic rupture was followed up by MRI or CT scan. Twenty-one patients in Group II underwent aortic repair an average of 8.6 months after the injury. In two patients the operation was expedited because of an enlarging aortic aneurysm. In Group I the postoperative mortality was 19%, three patients developed postoperative paraplegia and one acute renal failure. In Group II there were no postoperative deaths and no major complications. PMID- 10840206 TI - Anomalous origin of the right coronary artery from the left coronary sinus: case report and review of surgical treatments. AB - An anomalous right coronary artery arising from the left sinus of Valsalva is a rare but potentially lethal abnormality. We present a case report and literature review of this anomaly as well as its surgical management in the face of unobstructed distal coronary arteries. Furthermore we report the use of intraoperative transesophageal stress echocardiography to evaluate adequacy of graft flow. PMID- 10840207 TI - An unusual cause of femoral embolus. AB - Acute lower limb ischaemia is associated with high morbidity and mortality. The authors describe an unusual cause of arterial occlusion two weeks following aorto coronary bypass surgery. This case should make vascular surgeons aware of the possibility of prosthetic materials acting as emboli. Furthermore, they should also be mindful that the presenting symptom may be at a site and time remote from the original intervention. PMID- 10840208 TI - Lack of compressibility of the common femoral vein: an unequivocal sign of proximal deep venous thrombosis on duplex ultrasound? AB - The authors report a case of false positive diagnosis of iliac vein thrombosis on duplex ultrasound. The misreading of the duplex scan was based on the presence of an incompressible common femoral vein. External compression on the iliac veins, caused by enormous postoperative lymphoceles, was responsible for the false positive diagnosis. PMID- 10840209 TI - Chronic disseminated intravascular coagulopathy cured by abdominal aortic aneurysm repair. AB - Disseminated intravascular coagulopathy (DIC) may rarely be caused by a previously asymptomatic abdominal aortic aneurysm (AAA). The authors describe a recent case where repair of the AAA provided a complete cure for the patient's bleeding tendency. The multidisciplinary management of this patient is presented, and the evidence for the rare causal role of AAA in DIC is discussed. Coagulation disorders in aneurysm patients are probably under-reported, and warrant careful perioperative assessment. PMID- 10840210 TI - Toxic shock syndrome. AB - Toxic shock syndrome (TSS) is an acute febrile illness accompanied by watery diarrhea, sunburn rash, vomiting, hypotension, and multiple organ involvement. When it was initially recognized 20 years ago, TSS was primarily associated with menstruation. Since then, the frequency of menstrual TSS has decreased dramatically. Staphylococcus aureus is the cause of menstrual TSS and is also responsible for about half of all cases of nonmenstrual TSS. Since the late 1980s, Streptococcus pyogenes has been shown to cause a nonmenstrual TSS-like illness, with a higher mortality rate. The mechanism of TSS is attributable to bacterial endotoxins that impede the body's immune system and cause the release of cytokines, which are responsible for the clinical manifestations of the disease. Early recognition and treatment of TSS are important factors in improving patient survival. Patients should be treated with parenteral antibiotics that are highly effective against staphylococci and streptococci. They should also receive fluid resuscitation and vasopressors. Some patients require intubation, dialysis, or immunoglobulin therapy. PMID- 10840211 TI - Venous thromboembolic complications in obstetrics and gynecology with a focus on the role of low molecular weight heparin. AB - Venous thromboembolism is a major cause of morbidity, mortality, and economic expense in the field of obstetrics and gynecology. Without prophylaxis, as many as 30% of at-risk patients will suffer deep vein thrombosis, and nearly 1% of these patients will succumb to a fatal pulmonary embolism. Both deep venous thrombosis and pulmonary embolism manifest few specific symptoms, and the presentation of these entities is often clinically silent. Therefore, a prophylactic approach is preferred. We review the pathophysiology, risk factors, prophylactic choices, and treatment of venous thromboembolic complications for the general obstetric and gynecology population. We focus on the role of low molecular weight heparin because of its convenient dosing and favorable risk/benefit profile. PMID- 10840212 TI - Access and utility as reflections of cultural constructions of pregnancy. AB - Health care providers can give their patients better care if they understand how their patients view pregnancy and birth. This article provides some examples of how women from various cultural backgrounds understand pregnancy and how these beliefs affect women's decisions to seek prenatal care and to utilize prenatal services regularly throughout their pregnancies. These concepts-access and utility-provide the frame for this article, and the case studies from diverse ethnic groups provide examples of a variety of cultural beliefs and women's decisions to seek and to value biomedical direction during pregnancy. The conclusion includes several recommendations that health care providers can employ to enhance the quality and effectiveness of care. PMID- 10840213 TI - Diagnosis and treatment of bipolar disorder in pregnant women. AB - Bipolar disorder is relatively rare in obstetrics and gynecology (ob/gyn) practice compared with depressive and anxiety disorders, but there is a high risk for poor outcomes for patients and their offspring. Ob/Gyn physicians are assuming increasing responsibility for the care of these patients in today's managed care environment, working independently or in collaboration with a psychiatrist. The clinical presentation of bipolar patients may include mania and/or depression, in addition to more minor mood fluctuations that accompany the emotional and physical changes of pregnancy. The key issue in the differential diagnosis is to rule out medical, surgical, medication, and substance etiologies of mania that are potentially reversible. Indications for routine, urgent, and emergent referrals to a psychiatrist are reviewed. Treatment for bipolar women considering pregnancy includes prepregnancy planning education, involving the patient, family, psychiatrist, ob/gyn physician, and maternal-fetal medicine specialist. Treatment for pregnant bipolar women includes an individualized risk/benefit assessment regarding medication, monitoring levels and adherence to medication if it is used, ongoing patient education, and collaboration between the ob/gyn physician, maternal-fetal medicine specialist, and psychiatrist. PMID- 10840214 TI - When and how to use mood stabilizers during breastfeeding. AB - For many women with bipolar disorder, treatment with mood stabilizers is indicated to decrease the risk of a recurrence of depression, mania, or psychosis during the postpartum period. Because breastfeeding during this time exposes infants to these medications, a risk-benefit analysis is crucial for the mother and her physician when deciding whether to breastfeed. This article reviews the risks and benefits to mothers and nursing infants with regard to untreated mental illness, breastfeeding, and medication exposure during breastfeeding. Reports of breast milk and infant serum drug concentrations are summarized. Adverse events reported with lithium, valproate, and carbamazepine are also described. The current available data and their limitations are further summarized to allow physicians to assess more accurately the risks and benefits of the use of mood stabilizers during breastfeeding. Finally, a list of practical recommendations is provided. PMID- 10840215 TI - Cervical cancer screening in developing countries. AB - In developing countries, cervical cancer remains a clinical problem of public health proportions. Eighty percent of the approximately 400,000 new cases of cervical cancer each year occur in such settings. Primarily as a result of the introduction and success of screening programs, cervical cancer rates in developed countries have dropped remarkably. However, in most developing countries, screening programs are virtually nonexistent. Reasons for this relate primarily to lack of public awareness and the fact that screening via cytology is an untenable proposition for mass screening in such low-resource settings. Matching the resources available to provide cervical cancer screening in such settings requires alternative means of providing testing and treatment. Recent evidence indicates that, depending on local conditions and locally-made decisions, visual inspection of the cervix with acetic acid wash, coupled with a benign form of therapy, such as cryotherapy, could hold promise as a means of testing and treatment. By adapting resource-appropriate technologies and building local consensus regarding clinically driven public health approaches that are safe and feasible, the challenge of reducing the mortality from cervical cancer in developing countries can yet be met. PMID- 10840216 TI - Erectile dysfunction: a review of a common problem in rapid evolution. AB - Erectile dysfunction (ED) has been defined by the National Institutes of Health Consensus Conference in 1993 as the inability to achieve and/or maintain an erection adequate for penetration and completion of satisfactory intercourse.1 Erectile dysfunction, the preferred term, is more accurate and less pejorative than impotence.An estimated 20-30 million American men are affected with ED of varying degrees of severity. The Massachusetts Male Aging Study published in 1994 reviewed 1,211 men between the ages of 40 and 70; 52% reported ED with 9.6% having mild, 22.2% moderate, and 17.2% complete or severe ED.2 The National Health and Social Life Survey, authored by Laumann et al.,3 reviewed a population of men and women between the ages of 18 and 59. Of note, female sexual dysfunction exceeded male sexual dysfunction, with 43% of women complaining of sexual function problems. Interestingly, except for lubrication, this seemed to decrease with age in women. On the other hand, 31% of the men complained of sexual problems that increased with age.Erectile dysfunction is an age-dependent problem. Approximately 2% of men are affected at 40 years of age and about 25% or more at 65 years of age.4 However, ED is not an inevitable result of aging. Rather, as a man matures it is most likely that he will experience more of the neurovascular insults resulting in ED.The development of an erection and satisfactory sexual function is a complex process. As suggested by Melman et al.,5 "Erection is truly at least a sensory-motor-neuro-hormonal-vascular-psycho social-cultural-interpersonal event." There are two main classifications of ED, psychogenic and organic. Current thinking suggests that up to 80% of ED is primarily of organic etiology. Yet, there is always a psychogenic factor.6 Erectile dysfunction may signal serious underlying and potential life-threatening diseases, such as diabetes, hypertension, cardiovascular disease, peripheral vascular disease, and other neurologic and endocrine disorders. Therefore, questions regarding sexual function should be part of routine medical evaluation. PMID- 10840217 TI - Organic phosphates in the red blood cells of fish. AB - Fish are dependent on aerobic metabolism. They respond to changes in oxygen availability by a wide spectrum of compensatory and respiratory adjustments to safeguard tissue oxygenation. Such adjustments are directed to facilitate both oxygen uptake at the gas exchange surfaces and oxygen unloading to tissues. The importance of erythrocytic organic phosphates as regards oxygen transfer has been recognised since 1967 when the 'dramatic' effect of 2,3DPG on human haemoglobin was first reported. The present review examines the appearance of all the major erythrocytic organic phosphates during the evolutionary radiation of fish. In addition, it provides examples illustrating qualitative and quantitative ontogenetic changes of organic phosphates in the red blood cell of several fish species and describes their effects on oxygen affinities. The interaction of the organic phosphates with haemoglobins and divalent cations are also examined. Of particular interest is the regulation of erythrocytic organic phosphates according to both environmental and physiological conditions. PMID- 10840218 TI - Metabolic responses of the white shrimp, Penaeus vannamei, to ambient ammonia. AB - Juvenile shrimp were individually exposed during 24 h to 0.007 (control), 0.36, 1.07, and 2.14 mmol/l total ammonia-N at 28 degrees C and 39 ppt salinity. After 22 h of ammonia-N exposure, oxygen consumption was measured for 2 h, and then hemolymph, hepatopancreas, and muscle tissues were sampled. Oxygen consumption, and levels of lactate and glycogen in the hepatopancreas increased significantly at the highest ammonia-N concentration. Concentration of oxyhemocyanin, acylglycerol, and cholesterol in hemolymph, and lactate in muscle decreased significantly in the group exposed to the highest ammonia levels. The changes observed in hemolymph and tissue metabolic fuels suggest a reduced use of carbohydrate through anaerobic metabolism and an increase in the use of lipids to satisfy the metabolic demand. PMID- 10840219 TI - Protease activity in fractionated blood cells of the vanadium accumulating ascidian Phallusia mammillata. AB - Proteolytic activity was studied in the fractionated blood cells of the vanadium accumulating ascidian P. mammillata by separating the cells before measuring their activity. Cells were separated to avoid vanadocyte breakdown and subsequent vanadium diffusion into the assay medium. Our study revealed activity in the morula cell extract that was obtained by sonication and Centricon concentration. After removing part of the extract for enzyme activity assay the remainder was kept at 0 degrees C; it was later found that much of the protein in this latter fraction formed a sediment whereas the protease remained in solution. The serine protease substrate specificity of the enzyme was measured and the results are discussed in relation to serine protease involvement in immune defense. PMID- 10840220 TI - Comparisons of different stages of chick embryonic development by the physiological regulatory response to hyposmotic challenge. AB - Cardiac myocytes isolated and cultured from 11 day chick embryos present a Ca(2+) dependent regulatory volume decrease (RVD) when exposed to hyposmotic stimulus. The RVD of myocytes from different embryonic stages were analyzed to evaluate their physiological performance through development. Among the several embryonic stages analyzed (6, 11, 16 and 19 days) only 19 day cardiac myocytes present a greater RVD when compared with 11 day (considered as control), the other ages showed no difference in the regulatory response. As it is known that RVD is Ca(2+) dependent, we decided to investigate the transient free Ca(2+) response during the hyposmotic swelling of the 11 and 19 day stages. The 11 day cardiac myocyte showed a transient 40% increase in intracellular free Ca(2+) when submitted to hyposmotic solutions, and the free Ca(2+) returned to baseline levels while the cells remained in hyposmotic buffer. However, the intracellular free Ca(2+) transient in the 19 day cells during hyposmotic challenge increases 100% and instead of returning to baseline levels, declines to 55% above control, well after the 11 day transient has returned to baseline. Also, quantitative fluorescence microscopy revealed that 19 day cardiac myocytes have more sarcoplasmic reticulum (SR) Ca(2+) ATPase sites per cell as compared to the 11 day cells. Our findings suggest that 19 day cells have more developed intracellular Ca(2+) stores (SR). By evoking the mechanism of Ca(2+) induced Ca(2+) release, the cells have more free Ca(2+) available for signaling the RVD during hyposmotic swelling. PMID- 10840221 TI - Effect of somatostatin on electrogenic ion transport in the duodenum and colon of the mouse, Mus domesticus. AB - In this study, we have used the mouse intestine and the Ussing short circuit technique to compare the effects and mechanism of action of somatostatin (SST, 0.1 microM) on cAMP- and Ca(2+)-mediated ion secretion in the duodenum and colon of the Swiss-Webster mouse. The cAMP-dependent secretagogues, prostaglandin E(2) (1 microM) and dibutyryl-cAMP (150 microM) increased short circuit current (I(sc)) in both regions, but only the colonic response was inhibited by SST. This inhibition was independent of enteric nerves, suggesting a direct action on the epithelial cells. The Ca(2+)-dependent secretagogue carbachol (10 microM) stimulated a transient increase in I(sc) in both intestinal segments. In the duodenum, SST partially inhibited this increase in I(sc) and both the responses to carbachol and SST were independent of enteric nerves. In the colon, while SST inhibited the carbachol induced increase in I(sc), pre-treatment with tetrodotoxin (750 nM) profoundly inhibited the carbachol induced increase in I(sc), thus markedly reducing the inhibitory effect of SST. This indicates an involvement of the enteric nervous system in the response to carbachol and the action of SST in the colon. These data indicate marked regional differences within the mouse intestine of the effects of SST on ion secretion and demonstrate different mechanisms of action of SST in the duodenum and colon. PMID- 10840222 TI - Monoamines and their metabolites in the freshwater snail Biomphalaria glabrata. AB - Metabolism of the major monoamines and their functions were studied in the freshwater snail Biomphalaria glabrata. In both juvenile and adult snails, the plasma (cell-free hemolymph) appears to act as a reservoir for most of these monoamines and their metabolites including among others, L-dopa and dopamine as major constituents. Significant quantities of L-tryptophan, precursor of indoleamines, also was found in the plasma. L-dopa, serotonin, homovanillic acid and dopamine were prominently represented in the central nervous system of the snail, while serotonin and its metabolites, 5-hydroxyindole acetic acid and 5 hydroxytryptophol were found in the ovotestis. Catecholamines such as L-dopa, dopamine and homovanillic acid were identified in the albumen gland. Functional aspects of both dopamine and serotonin were studied using in vitro cultures of albumen glands, the site of perivitelline fluid and galactogen synthesis in B. glabrata. Dopamine was found to stimulate the release of secretory proteins when exogenously added to gland cultures and this process was inhibited by chlorpromazine, a dopamine receptor antagonist. Similarly, exogenous serotonin stimulated in vitro protein secretion by albumen glands. Thus, these results suggest that monoamines may play important roles in regulating reproductive activity of this snail and provides an excellent model for studying neurotransmitter function and metabolism in molluscs. PMID- 10840223 TI - Demineralization of the vertebral skeleton in Atlantic salmon Salmo salar L. during spawning migration. AB - In Atlantic salmon, Salmo salar, the mineral rate of vertebrae in a given fish varies according to the position of the vertebra along the rachidian axis. Indeed, the mean rate goes from 49% in the anterior vertebrae and raises to 51% in post-truncal vertebrae. Although no significant difference in the mineral rate was noticed between males and females either in the lower river basin or after spawning, the mineral rate of vertebral bone decreased significantly (1-2%) during spawning migration. Vertebrae, like scales, are an important reservoir of calcium from which fasting salmon draws the minerals and organic materials necessary for the substantial remodeling of cranial bones in males and for sexual maturation. We hypothesize that mineral decrease in vertebrae may be the result of a halastasic demineralization of the vertebral tissues. PMID- 10840224 TI - C-reactive protein in the hemolymph of Achatina fulica: interrelationship with sex steroids and metallothionein. AB - C-reactive protein in Achatina fulica (ACRP) is a normal component of the hemolymph. Its concentration varied from 1mg/ml in the newly hatched male, 3-5 mg/ml in the most active hermaphrodite and 1.5-2.8 mg/ml in the sedentary female showing a direct relationship of the protein with the active phase of the animal. ACRP has a molecular mass of 400 kDa and showed high absorbance in the region of 200-230 nm. It has four subunits with relative molecular masses of 110, 90, 62 and 60 kDa, respectively. Interestingly, rat platelet aggregation in vitro was significantly enhanced by ACRP in presence of 10 microM ADP and 2 mM Ca(2+) suggesting a probable role of ACRP in the aggregation of amoebocytes during the formation of plug in injured tissue. Like other vertebrate CRPs, ACRP also acts as a scavenger of chromatin fragments as evidenced by its binding to poly-L arginine. Among the sex steroids, 4-androstenedione induces ACRP synthesis in the newly hatched male reaching the level found in the most active hermaphrodite phase (4 mg/ml). A very high molar ratio (5) of mercury binding to ACRP confirmed its sequestration property of heavy metals as observed in vertebrates. The level of metallothionein (MT) in the hemolymph gradually increased from the male to the hermaphrodite to the female, a pattern distinctly different from that of the ACRP titer. Since both MT and ACRP can sequester inorganic mercury, the high level of MT compensates functionally for the low titer of ACRP in the sedentary female. PMID- 10840225 TI - Use of isolated digestive-gland cells in the study of biochemical and physiological processes in gastropod molluscs. AB - We describe a method for preparation and maintenance of isolated digestive-gland cells in the abalone, Haliotis kamtschatkana. Viability of the isolated cells was confirmed by the fact that 18 h after preparation the cells exhibited less than 5% staining with trypan blue and actively synthesized glycogen following the addition of glucose substrate. Use of the method in a 15-month study of metabolic activity of the digestive gland of H. kamtschatkana showed significant differences in oxygen consumption of isolated-cell preparations correlated with seasonal differences in somatic and gametogenetic growth, and with relative size of the digestive gland. PMID- 10840226 TI - Heterogeneity in growth and differentiation characteristics in male and female satellite cells isolated from turkey lines with different growth rates. AB - The effects of growth- and gender-related differences on satellite cell proliferation and differentiation were investigated using satellite cells isolated from the pectoralis major muscle of a turkey line selected for increased 16-week body weight (F-line) and its unselected randombred control (RBC2-line). Proliferation rates within the F- and RBC2-lines did not differ between sexes. The F-line male and female satellite cells when compared to the RBC2-line male and female satellite cells proliferated at a faster rate. Differentiation rates were increased for the F-line male cells compared to both the F-line female and RBC2-line male satellite cells. No difference in differentiation rate was noted within the RBC2-line satellite cells. For satellite cells from females, the RBC2 line differentiated faster than the F-line. Morphological data on myotube length and the number of nuclei per myotube supported the differentiation data in that F line male satellite cells had the longest myotubes with the most nuclei, there was no significant difference between myotubes within the RBC2-line, and female derived myotubes from the RBC2-line were longer than those of the F-line by 96 h of fusion. These data are suggestive of both growth- and gender- related differences in satellite cell proliferation and differentiation. PMID- 10840227 TI - Species specificity of barnacle settlement-inducing proteins. AB - We previously isolated a larval settlement-inducing protein complex (SIPC) from adult extracts of the barnacle, Balanus amphitrite using a nitrocellulose membrane settlement assay. In the present study, we found that the extracts of other adult barnacles, Megabalanus rosa and Balanus eburneus, also induced the settlement of B. amphitrite cyprids although the inductive activity was slightly lower than that of conspecific extracts. Furthermore, we examined reactivity to anti-SIPC antibody in adult extracts from six species of Japanese barnacles other than B. amphitrite, brine shrimp and eight marine sessile organisms besides barnacles. The results showed that all barnacles examined contained SIPC-like proteins with slightly different molecular weight, while the other animals did not react to the antibody by immunoblot analysis. These findings suggest that species specificity in settlement-inducing proteins of barnacles is not so strict, but these proteins are characteristic to barnacle species. PMID- 10840228 TI - Model analogues in the study of cephalic circulation. AB - Simple laboratory models are useful to demonstrate cardiovascular principles involving the effects of gravity on the distribution of blood flow to the heads of animals, especially tall ones like the giraffe. They show that negative pressures cannot occur in collapsible vessels of the head, unless they are protected from collapse by external structures such as the cranium and cervical vertebrae. Negative pressures in the cerebral-spinal fluid (CSF) can prevent cerebral circulation from collapsing, and the spinal veins of the venous plexus can return blood to the heart in essentially rigid vessels. However, cephalic vessels outside the cranium are collapsible, so require positive blood pressures to establish flow; CSF pressure and venous plexus flow are irrelevant in this regard. Pressures in collapsible vessels reflect pressures exerted by surrounding tissues, which may explain the observed pressure gradient in the giraffe jugular vein. Tissue pressure is distinct from interstitial fluid pressure which has little influence on pressure gradients across the walls of major vessels. PMID- 10840230 TI - Editorial PMID- 10840229 TI - Short chain fatty acids but not lactate or succinate stimulate mucus release in the rat colon. AB - BACKGROUND: Short chain fatty acids (SCFAs) affect various intestinal functions. Mucus is an important physiological component of the intestinal mucosal barrier. However, the effect of SCFAs or other organic acids on the intestinal mucus release is poorly understood. The aim of this study was to investigate whether lumen SCFA stimulates mucus release into the rat colon. METHODS: A solution of SCFA, lactate or succinate was infused into the colon of anesthetized rats, and we then measured the hexose content of the effluent. We also examined the influence of cholinergic antagonists on the effects of SCFA. RESULTS: A SCFA mixture (75 mM acetate, 35 mM propionate and 20 mM butyrate) or individual SCFAs (130 mM) increased the mucus release into the colon in a similar manner. The individual SCFAs, but not lactate or succinate, stimulated colonic mucus secretion in similar concentration-dependent manners. Butyrate stimulated colonic mucus secretion at 20 mM, but acetate, propionate, lactate and succinate at this concentration did not. Pretreatment with an anti-cholinergic agent diminished the stimulatory effects of SCFAs on mucus secretion. CONCLUSIONS: Lumen SCFAs, but not lactate or succinate, stimulate mucus release from the rat colon via a cholinergic nerve mechanism. PMID- 10840231 TI - This month in jan PMID- 10840232 TI - Barriers to learning from reflection: a study of the use of groupwork with post registration nurses. AB - There are few studies which describe the use of groups to facilitate reflection and even fewer which evaluate the effectiveness of such groups. Much of the literature discussing the techniques used to facilitate reflection suggests that learners willingly engage in such processes and find them enjoyable. This paper reports on a qualitative study of the use of groups to develop learning through reflection on a part-time post-registration diploma programme for nurses. The findings suggest that there are many barriers to learning which must be overcome before practitioners can use the opportunities to reflect on and learn from their experience. It is also contended that such learning may not necessarily be an enjoyable experience. The findings suggest that previous educational experience and the current culture in which nurses and midwives work, impose tremendous barriers to reflecting on, and learning from, experience. In particular these affect the willingness of learners to expose themselves to the judgement of others and their ability to be open to taking responsibility for their own learning. Other barriers to learning in groups are the effects of other group members, their commitment or resistance to shared learning, the ways in which group members interact with each other and facilitation styles. PMID- 10840233 TI - Reflection: nursing's practice and education panacea? AB - Nurses are now directed to 'reflect effectively and practice reflectively'. When there is scant evidence to support the use of reflection, why does the United Kingdom Central Council, English National Board for Nursing Midwifery and Health Visiting and institutes of nurse education insist that nurses, at all levels of experience, reflect? Before developing the debate in relation to the robustness or otherwise of the concept of 'reflection' and its application to practice, a discussion as to what constitutes nursing knowledge, followed by an examination of the main theories related to adult learning, is presented. The paper develops to encompass a critical examination of both theoretical and applied perspectives on reflection. Conceptual issues and assumptions related to reflection are explored. Key areas considered are whether the practice of reflection facilitates the improvement of one's knowledge, helps to generate theory and results in one becoming a more critical practitioner with resulting benefits for patients. It will be argued that reflective theory and practice has not yet been adequately tested and that there is a pressing need for evidence to demonstrate irrefutably the effectiveness of reflection on nursing practice, particularly with respect to patient outcomes. PMID- 10840234 TI - Pedagogical implications of approaches to study in distance learning: developing models through qualitative and quantitative analysis. AB - The need for flexibility in the delivery of nurse education has been identified by various initiatives including: widening the entry gate; continuous professional development; and the specialist practitioner. Access to degree level programmes is creating the need to acquire academic credit through flexible learning. The aim of this study was to further develop relationships between the need for guidance, materials design and learning styles and strategies and how these impact upon the construction of meaning. The study is based on interviews of 20 female community nurses purposively selected from the 96 respondents who had previously completed a survey questionnaire. The interviews were underpinned by theories relating to learning styles and approaches to study. Of particular concern was how these variables are mediated by student context, personal factors and materials design, to influence the need for support and guidance. The interview transcripts were first analysed using open and axial coding. Three approaches to study emerged from the data - systematic waders, speedy-focusers and global dippers - which were linked to other concepts and categories. Categories were then assigned numerical codes and subjected to logistical regression analysis. The attributes of the three approaches to study, arising from both qualitative and quantitative analysis, are explained in detail. The pedagogical implications of the three approaches to study are explained by their predicted relationships to other variables, such as support and guidance, organization of study, materials design and role of the tutor. The global dipper approach is discussed in more detail due to its association with a variety of predictor variables, not associated with the other two approaches to study. A feedback model is then developed to explore the impact of guidance on the global dipper approach. The paper makes recommendations for guidance to students using different approaches to study in distance learning. PMID- 10840235 TI - Neurological rehabilitation nursing: a review of the research. AB - Neurological rehabilitation is increasingly seen as a specialist form of rehabilitation requiring specialist nursing knowledge. However, as in many areas of nursing, nurses in this field recognize that there is a need to increase their knowledge base to ensure that they provide the best and most up-to-date quality care for their patients and their families. To achieve high levels of competence, neurological rehabilitation nurses need to be aware of the existing body of research in this field so that they can make sound decisions regarding their practice and future research endeavours. This review aims to validate the existing knowledge base in this area by identifying and critically analysing research conducted in the area of neurological rehabilitation nursing during the decade 1988-1998 and by identifying any gaps and weaknesses that may be addressed by future research. Studies were included in the review if they examined an aspect of rehabilitation nursing, were concerned predominately with the neurological patient and were conducted in an in-patient setting. Research with a medical focus was not included and only research published in English was reviewed. Relevant research located examined the following areas: (1) the role of the rehabilitation nurse; (2) specific problems concerning the neurological rehabilitation patient; (3) specific nursing interventions; (4) rehabilitation nursing practice; (5) the delivery of rehabilitation programmes and team working; and (6) the family of the rehabilitation patient. The review concludes that research in this speciality remains limited both in quantity and quality. Most of the studies lack rigour and, because they were conducted at a single site, their generalizability to other clinical sites may be limited. PMID- 10840236 TI - The evolving educational needs of nurses caring for the older adult: a literature review. AB - Recent changes in long-term care policies in the United Kingdom have resulted in many more older patients/clients, previously nursed in long-term hospital facilities, now being cared for in the community. This change has had a significant impact on nurses, forcing many to make the transition from working in hospital to within the community. This transition calls for appropriate professional educational preparation to enable these nurses to undertake their new roles effectively. The literature search that forms the basis for this paper revealed relatively little material focusing specifically on the educational needs of such nurses in transition. However, literature that addresses the needs of nurses caring for the older individual in the acute setting, and in the community environment, was found and is explored. PMID- 10840237 TI - The incidence of smoking amongst nurses: a review of the literature. AB - Over the years there has been an ongoing debate about the implications of the smoking behaviour of nurses in relation to their own health and in the context of their professional role and responsibilities. However it is difficult to determine with any degree of accuracy the current incidence of smoking amongst nurses and most of the available research is based on small samples which embrace nurses of different levels and status. Very few recent studies in the United Kingdom or elsewhere have focused on this issue. A confused picture therefore emerges. This paper presents a critical overview of the literature and where appropriate provides data about the incidence of smoking in comparable groups of the general population. PMID- 10840238 TI - Factors associated with students' orientations to nursing. AB - Factors associated with students' orientations to nursing This paper presents the results of a study focusing on the factors associated with orientations to nursing. Students' orientations to nursing have not as yet been a focus of nursing research. In some other professions, however, professional orientation has been associated with learning motivation and study performance, and has been seen as a predictor of work satisfaction. In this study, students' orientations to nursing were defined in terms of caring, nursing expertise and life orientation. The hypothesis of whether students' pre-educational experiences of nursing, gender, choice of nursing specialty, problems with nursing studies and intention to stay in nursing were associated with different orientations was tested. The extent to which students were orientated to caring, nursing expertise and their own life was also examined. The orientation to nursing measurement tool, which has been developed on the basis of a qualitative study, was used to collect the data. Nurse teachers collected the data from nursing students (n=184) who were studying in three different nursing programmes in Finland. Non parametric assessments (Mann-Whitney U-test and Kruskal-Wallis test) of the differences between the students' orientations were carried out. A majority of the students were highly life-orientated, and two-thirds had average nursing expertise or caring orientation scores. The results supported the study hypothesis of an association between students' orientations and their gender, choice of nursing speciality, problems with nursing studies and intention to stay in nursing. However, the hypothesis of an association between students' pre educational nursing experiences and orientation to nursing was not supported. The contradictions between students' orientation to nursing and the philosophy of nursing underlying the study programme may be a source of motivational problems and dissatisfaction with nursing education. Therefore, nurse educators are challenged to discuss curriculum matters and student supervision in order to promote flexibility in planning personal study programmes. PMID- 10840239 TI - School nursing: costs and potential benefits. AB - BACKGROUND: Previous reports that variations in school nursing resources across the UK had no relationship to deprivation; controversy about the changing role of the school nursing service. OBJECTIVES: To measure the resources allocated to school nursing, determine whether the variations can be explained by deprivation, and assess whether the allocation of school nursing time to a range of tasks is in line with current evidence and perceptions of changing needs. STUDY DESIGN: Quantitative economic analysis; qualitative descriptive study. SETTING: Detailed study of four English districts with diverse characteristics; staffing and service questionnaire and telephone survey of 62 districts. MAIN MEASURES: Staff resources and their salaries; measures of population and deprivation; activity statistics. RESULTS: There were wide variations in the cost of the school nursing service, but in contrast to previous reports 24% of the variance was explained by deprivation. There were no clear associations with any other social or educational variables. The greatest allocation of time was in routine screening and surveillance tasks. Relatively little time was allocated to other activities such as health promotion, support of special needs or unwell children, or teenage clinics. The expenditure on school nursing is only loosely related to deprivation and the results of this study offer guidance on what districts should spend to achieve equity of provision. CONCLUSIONS: The current allocation of resources to school nursing in between districts comparisons is not equitable and the use of school nursing time is out of step with current evidence of need and effectiveness. PMID- 10840240 TI - Fit for purpose: the relevance of Masters preparation for the professional practice of nursing. A 10-year follow-up study of postgraduate nursing courses in the University of Edinburgh. AB - Continuing education is now recognized as essential if nursing is to develop as a profession. United Kingdom Central Council for Nursing, Midwifery and Health Visiting (UKCC) consultations are currently seeking to establish appropriate preparation for a 'higher level of practice' in the United Kingdom. The relevance of Masters level education to developing professional roles merits examination. To this end the results of a 10-year follow-up study of graduates from the Masters programme at the University of Edinburgh are reported. The sample was the entire cohorts of nurses who graduated with a Masters degree in the academic sessions from 1986 to 1996. A postal questionnaire was designed consisting of mainly closed questions to facilitate coding and analysis but also including some open questions to allow for more qualitative data to be elicited. The findings indicated clearly that the possession of an MSc degree opened up job opportunities and where promotion was not identified, the process of study at a higher level was still perceived as relevant to the work environment. This applied as much to the context of clinical practice as to that of management, education or research. The perceived enhancement of clinical practice from a generic Masters programme was considered a significant finding. Also emerging from the data was an associated sense of personal satisfaction and achievement that related to the acquisition of academic skills and the ultimate reward of Masters status. The concept of personal growth, however, emerged as a distinct entity from that of satisfaction and achievement, relating specifically to the concept of intellectual sharing, the broadening of perspectives and the development of advanced powers of reasoning. PMID- 10840241 TI - Managing the unmanageable? Nurse Executive Directors and new role developments in nursing. AB - This paper offers a unique insight into the factors affecting the functioning of the Nurse Executive Director(s) and their views on the realities of nursing management in the new National Health Service in England. It is based on the findings of the Exploring New Roles in Practice (ENRiP) project which was carried out for the Department of Health. As part of this project interviews were undertaken with Nurse Executive Directors from a 20% sample of acute hospital trusts in England. The interviews were designed to explore the Nurse Executive's perceptions of the process of new role development. The findings highlight a tension between the drive for professional development and the pragmatics of service delivery in a health care system which lacks dedicated funding for nursing development. It was evident that most Nurse Executives vacillated between responding either in an ad hoc way to internal and external pressures or leading and supporting managed development. PMID- 10840242 TI - Blurring boundaries: professional perspectives of the emergency nurse practitioner role in a major accident and emergency department. AB - The role of the emergency nurse practitioner (ENP) has increasingly become part of mainstream healthcare delivery in major accident and emergency departments in the United Kingdom. Although some research data are available in this field, there has been little attempt to evaluate the impact of the implementation of the ENP role from the perspective of those healthcare professionals most closely involved at local level. This paper describes one part of a case study evaluation of the role in an accident and emergency department in the South Thames English region. Nine face-to-face semi-structured interviews were carried out with the key multidisciplinary stakeholders in the organization. Five major themes emerged from the data analysis: blurring role boundaries; managing uncertainty; individual variation; quality vs. quantity; and the organizational context. Whilst some professional consensus was evident regarding the benefits of the role, such as improved waiting times and patient satisfaction, there appeared also to be a degree of ambivalence, particularly regarding current role configuration, value for money, and the extent to which the role should be expanded in the future. These issues are discussed in terms of professional identity, changing role boundaries, and professional personhood. It is argued that the benefits and pitfalls of the ENP role need to be considered within the context of local service provision. The growing emphasis on clinical governance reinforces the need for ongoing audit of role effectiveness in order to meet the challenges and uncertainties of increasingly blurred professional boundaries. PMID- 10840243 TI - A review of health professional attitudes and patient perceptions on 'inappropriate' accident and emergency attendances. The implications for current minor injury service provision in England and Wales. AB - Increasing attendances in accident and emergency (A and E) departments in the United Kingdom have been attributed to a greater number of patients presenting with minor injuries. A and E staff believe this type of patient is suitable for primary care, and is 'inappropriate' for A and E management. Thus, A and E staff find 'inappropriate' attenders time-consuming and unrewarding, and are less motivated to help them, whilst 'inappropriate' patients believe they have attended the appropriate service for their medical needs and expectations. This review examines research into health professional and patient attitudes towards 'inappropriate' attendances in accident and emergency. It identifies a discrepancy between health professional and patient perspectives regarding 'inappropriate' attendances. However, the change in accident and emergency services with the development of minor injury units and nurse practitioners within A and E to treat minor injury patients away from the mainstream A and E service, appears to be based on the professional attitude of what constitutes an appropriate A and E attendance, and not on the patients' perspective. As negative attitude formation towards 'inappropriate' A and E attendances has occurred, there is concern that such attitudes could remain or develop again in the new units. Patients are generally not medically trained and may experience difficulty in ascertaining the severity of their own condition and attending the 'appropriate' service, as defined by trained professionals. This is exacerbated by the unclear boundaries and roles of minor injury units, nurse practitioners and general practitioners in minor injury care. Therefore research is required into current attendances in minor injury units, A and E departments and general practice, in order to develop clear roles and boundaries for these services. More importantly, research is warranted into the attitudes of all minor injury care providers towards attending patients, and into patient perceptions of the services offered. PMID- 10840245 TI - Clinical effectiveness in the post-anaesthesia care unit: how nursing knowledge contributes to achieving intended patient outcomes. AB - Clinical effectiveness in the post-anaesthesia care unit: how nursing knowledge contributes to achieving intended patient outcomes This paper reports part of an ongoing study on how nursing knowledge develops in practice and influences patient outcomes. The practice focus is post-anaesthesia nursing, an area which has been under-represented in nursing research. A qualitative approach was used to explore narrative data, collected by in-depth interviews, with 32 experienced post-anaesthesia nurses. Data analysis and interpretation were informed by a phenomenological perspective consistent with a research focus on individual experiences. Interpretation of the narratives gave rise to a description of knowledge development which occurred as nurses gained experience, drew on the available knowledge base, and used knowledge in practice. This led to the identification and description of 'referential' and 'effective' types of knowledge. Referential knowledge comprises the variety of external knowledge sources available to inform practice. When referential knowledge was incorporated into the personal knowledge of the individual nurse in everyday practice, it was transformed into effective knowledge which was used by nurses to achieve desired patient outcomes. The study findings indicate that nurses used effective knowledge as evidence for decision making and interventions in everyday practice. The key finding of the study is that effective nursing knowledge requires practice for its development. PMID- 10840244 TI - Nursing scripts and the organizational influences on critical thinking: report of a study of neonatal nurses' clinical reasoning. AB - During 1995-1997 a study was undertaken to explore the extent to which theoretical knowledge acquired through a distance education programme in neonatal nursing was brought to bear in the real-world clinical reasoning of course participants. The study utilized a think aloud technique and included both concurrent (on-the-job) and retrospective verbal reports at 0, 6 and 12 months into the programme. Participants (n=4) were also interviewed individually on completion of the study. Results indicated that important inconsistencies existed between participants' theoretical knowledge and their practice; they also pointed to some organizational influences on these theory-practice inconsistencies. Script (or schema) theory provided a useful explanatory framework for these results. The paper includes a brief description of data collection and analysis techniques; its main emphasis, however, is on these theory-practice inconsistencies and their explanation in terms of the nature and acquisition of nursing practice scripts. The implications of nursing scripts for the promotion of critical thinking and evidence-based practice are discussed. PMID- 10840246 TI - Exploring reflective thinking in nursing practice. AB - While it is claimed in the nursing literature that reflective thinking is the approach par excellence for learning and advancing the art and practice of nursing, few empirical studies have been undertaken in this area to date. Sense Making, a qualitative research method, was utilized to obtain and analyse data from interviews with 10 registered nurses in order to study reflective thinking in actual nursing practice. Ten non-routine nursing situations were analysed for the presence of reflective thinking. Time-line interviews of the events resulted in a total of 59 micro-moments, each of which was explored in terms of the thinking processes utilized to make sense of the situation as well as the focus of their thought. 'Pre-perceptions' played an important part in how the respondents perceived their situation. Reflective thinking was extensively manifest, especially in moments of doubt and perplexity, and consisted of such cognitive activities as comparing and contrasting phenomena, recognizing patterns, categorizing perceptions, framing, and self-questioning in order to create meaning and understanding. Self-questioning was identified as a significant process within reflective thinking. By exploring and analysing the type of questions respondents were asking themselves, the study uncovered three hierarchical levels of reflective thinking. Respondents most often engaged in reflective thinking-for-action which centred on the here and now in order to act. Reflective thinking-for-evaluation focused on creating wholeness and contributed to the realization of multiple perceptions and multiple responses. Reflective thinking-for-critical-inquiry could not be demonstrated in the study sample. The findings of this study resulted in the development of a model of reflective thinking, which is discussed in terms of the implications for learning in nursing practice. PMID- 10840247 TI - Alteration to will as an experience of technology and nursing. AB - This paper presents one finding to arise from a recent qualitative research study that examined ways of understanding technology in surgical nursing, and examines its implications for nursing and health care practice. Although the research reported identified eight qualitatively different ways of understanding technology, this paper examines the specific experience that technology can alter will (volition). The experience described is a new area of knowledge and is in need of further examination and research. Nonetheless, it is significant to understanding technology, contemporary nursing practice and the provision of health care services. The paper concludes with some suggested approaches for curtailing the experience and a discussion related to challenges that arise from the finding that technology can alter the free will of nurses. PMID- 10840248 TI - Factors affecting the administration of analgesia to patients following repair of a fractured hip. AB - Fracture of the neck of femur, or hip, has become common in older females and the resulting pain is often a factor in inhibiting early mobilization following surgical repair. Since the administration of analgesia is dependant upon decision making of the nurse, this study concentrated on what influences nurses in the administration of analgesia to patients following surgical repair of a fractured hip. A review of the literature identifies the main influencing factors as nurses' lack of knowledge and the drug prescription, and suggests that education is the key enabler. However, much of this research is limited to controlled situations outside the clinical setting with a focus on the internal processes of decision-making. This ethnographic multiple-case study aimed to analyse factors which influence nurses' decision-making in the clinical setting and define behaviours in context. Following periods of participant observation and observation of documentation, semi-structured interviews were conducted to explore further the rationale behind nurses' decisions. Factors found to influence decision-making were: time, organization of care, influence of shift worked, impact of the multidisciplinary team (MDT), concerns over the use of opioid analgesia, and information giving and collection. These influences were found to be dependant on location (ward), and shift, with the factor of time providing a tension between all influences. The study findings suggest that education alone will not improve the administration of analgesia. It recommends that the voice of patients is heard more clearly, that the administration of analgesia is separated from the routine drug round, that attention is paid to how decision-making by nurses is supported in clinical settings, and that education be organized on a multidisciplinary basis. In addition, further research should be carried out in the clinical setting. PMID- 10840249 TI - The development of pressure ulcers in patients with hip fractures: inadequate nursing documentation is still a problem. AB - The aims of the study were to investigate, on a daily basis: (i) the development and progress of pressure ulcers, (ii) the documented nursing interventions for prevention and treatment of pressure ulcers, and (iii) when nursing interventions regarding prevention and treatment of pressure ulcers were documented, in relation to patient risk status and the development of pressure ulcers. The study design was prospective, comparative and descriptive. A total of 55 patients with hip fracture were included. To facilitate the nurse's assessment, a 'pressure ulcer card' was developed, consisting of the Modified Norton Scale (MNS) and descriptions of the four stages of pressure ulcers. The incidence of pressure ulcers was 55%. The mean rank of the lowest MNS score was significantly lower for patients who developed pressure ulcers than for patients without pressure ulcers. The majority of the pressure ulcers occurred between admission to the ward and the fourth day after surgery. Documented interventions regarding prevention and treatment were: repositioning, overlays, cushions, use of lotion and observation. The mean number of interventions per patient was 2.2 for patients who developed pressure ulcers during their hospital stay. The comprehensiveness and quality of the nursing record was unsatisfactory, and only three nursing records reached the level required by Swedish law. Preventive interventions such as repositioning were documented when the pressure ulcer had already occurred. The lack of nursing documentation regarding prevention and treatment of pressure ulcers may indicate that nurses did not identify pressure ulcers as a prioritized nursing problem for this patient group. The Modified Norton Scale could be a valuable tool for nurses, both identifying the patient at risk and acting as a guide for nursing interventions. The study was approved by the ethics committee of the Faculty of Medicine at Uppsala University. PMID- 10840250 TI - Managing post-discharge care at home: an analysis of patients' and their carers' perceptions of information received during their stay in hospital. AB - This small-scale study carried out in a Melbourne metropolitan hospital explored patients' and their carers' perceptions of information, adequacy of information, and their utilization of information concerning post-discharge care received from health professionals during their stay in hospital. The research design consisted of two stages. Stage one involved a qualitative approach using focused interviews of five pairs of patients and their carers, 2 weeks after discharge from hospital. Five main themes emerged from the content analysis of the interview transcripts: information given by health professionals to patients and carers, patients' and carers' psychological well-being, activities of daily living, caring tasks of the patients, and community linkages. A quantitative approach was used for stage two involving two sets of questionnaires, one for the patient and one for the carer, developed from the themes identified in stage one. A pilot study was conducted on three pairs of patients and their carers, 2 weeks after discharge from hospital. The main study consisted of a convenience sample of 40 pairs of patients and their carers who completed the questionnaires 2 weeks post discharge. Data analysis of stage two of the study consisted of descriptive statistics and cross-tabulations. The main findings suggested that carers received very little information from health professionals concerning their patients' health problems and care at home. The carers' health and employment states were often not considered in their patients' discharge plan. Carers who were present with their patients when they received information concerning post discharge care experienced a decrease in anxiety during their patients' convalescence at home, greater satisfaction with the information they received, and their patients experienced fewer medical problems post-discharge. The implications for nursing practice and research include recommendations for a more effective system of discharge planning, and further research to include a larger population with a more varied group of participants. PMID- 10840251 TI - An evaluation of information cards as a means of improving communication between hospital and primary care for women with breast cancer. AB - This paper describes an intervention study aimed at improving communication between hospital services and the primary health care team. A series of information cards were developed by breast specialist secondary care professionals for members of the primary health care team. Women with breast cancer were involved in the communication pathway and were asked to take the information cards to their own general practitioner (GP) practice. It was envisaged that women would be more likely to utilize the primary health care team for information if they were aware that the primary health care team was in receipt of information specific to the treatment they had received. Women newly diagnosed with breast cancer were allocated to either an intervention (n=38) or non-intervention (n=38) group. Patient interviews were carried out around the time of diagnosis and at 4 months from diagnosis. Interviews were also carried out with 31 GPs to ascertain their views on the provision of information for women with breast cancer, and on the information cards if relevant. The study findings were interesting although not significant in terms of the direction anticipated. The cards did not impact on the utilization of the primary health care team and women in the intervention group were no more likely to utilize primary care sources of information than women in the non-intervention group. Factors such as the long-standing relationship women had with their GP, the perceived lack of specialist knowledge on the part of GPs and district nurses, and the women's perception that information seeking was not a tangible reason for primary care contact had an impact on information-seeking behaviour. PMID- 10840252 TI - New pharmacological treatments for Alzheimer's disease: implications for dementia care nursing. AB - This paper addresses developments regarding new pharmacological treatments for people with Alzheimer's disease such as Donepezil and Rivastigmine. Following a brief discussion of the experience of people in the early stages of Alzheimer's disease, the pharmacology associated with these new treatments is outlined. The implications of these treatments for nurses working with this client-group are fully discussed within the context of assessment, treatment and cessation. It is argued that nurses have an important contribution in the management of these treatments and recommends that descriptive and evaluative research should be undertaken relating to this new specialist area of nursing care. PMID- 10840253 TI - Characteristics of severely mentally ill patients in and out of contact with community mental health services. AB - Community mental health nurses have been criticized for failing to prioritize work with people with a severe or enduring mental illness, many of whom have no contact with specialist mental health services and rely entirely on their general practitioner and primary care team. It is important to ensure that those in contact with specialist services actually need this level of input but, conversely, that those in contact with only primary care receive a service that is equipped to meet their needs. This study examines the differences between these two groups of patients. A sample of 253 community-based patients with a severe or enduring mental illness was divided into those with mental health service contact and those without. Differences in the demographic characteristics of the two groups were assessed and further, more detailed, comparisons were made with a sub-sample of 49 individuals randomly drawn from this larger sample. Instruments adopted for these purposes comprised the Camberwell Assessment of Need and the Life Fulfilment Scale. It was found that patients with a psychotic disorder were more likely than those with neurosis to be in contact with mental health services and patients with schizophrenia were significantly more likely to be on the active caseload of a community mental health nurse than those from all other diagnostic groups. However, patients' levels of need, unmet need and quality of life did not differ in relation to their service contact. Whilst the study provides limited evidence that community mental health nurses are targeting people with the most serious disorders, questions remain about the large proportion not receiving specialist care. Because primary care plays a significant role in the care of severely mentally ill people living in the community, the further mental health training of general practitioners and practice nurses is becoming increasingly important. PMID- 10840254 TI - Beliefs about voices and their effects on coping strategies. AB - Cognitive behavioural techniques are increasingly used as adjuncts to medication in the treatment of auditory hallucinations for people with schizophrenia. There are now literally hundreds of nurses trained in the use of cognitive behavioural interventions for psychosis. However, there is still disagreement about the nature of the cognitive processes that lead to deficits or biases in patients' processing of information about their psychotic experiences. Using Chadwick & Birchwood's Beliefs About Voices Questionnaire (BAVQ), the investigator collected data regarding voices from a sample of men and women being treated for schizophrenia by secondary mental health services. The investigator then carried out a cross-lagged panel analysis of the data. The investigator found, as predicted, positive relationships between a resistive coping style and an attribution of malevolence to voices, and between an engaging coping style and an attribution of benevolence to voices. Coping and attributional styles were not necessarily stable over time. There was a non-significant difference between women's and men's attributions and coping styles. There was less fluctuation over time in the women's scores on the BAVQ. This research shows that one cannot assume that either coping or attributional style becomes more stable over time. However, while there are strong relationships between attributions and coping styles, and particularly between malevolence and resistance and benevolence and engagement, these relationships are not necessarily mutually exclusive and some people in the study believe their voices to be both malevolent and benevolent. These findings suggest that clinicians need to make a very careful assessment of attribution and coping with regard to hallucinations and that systematic reassessment is very important. Further research is necessary in both the phenomenology of attribution and coping, but also to relate these variables to other aspects of schizophrenic illnesses. PMID- 10840255 TI - Enabling young 'ventilator-dependent' people to express their views and experiences of their care in hospital. AB - Discussion in this paper is drawn from a phenomenological study carried out in England describing the views and experiences of young 'ventilator-dependent' people aged 6-18 years, regarding their health and social care, education, and aspirations for the future. This study presented many methodological and ethical challenges in order to elicit and present their views in a meaningful way. Data were elicited through focused face-to-face interviews with 18 young people. Innovative methods of data collection were used with younger children and those who had a range of communication impairments. In addition, family members of 15 young people were interviewed. Presentation of findings is limited to views and experiences of the young people's care in hospitals. Findings reveal that a significant number of Articles in the United Nations (UN) Convention on the Rights of the Child were not respected or upheld. Young 'ventilator-dependent' people were discriminated against when trying to access health services because of their need for assisted ventilation. They were particularly excluded from making important decisions about their lives and were not always offered the full protection of the Children Act 1989 (England). Some young people were not able to maintain adequate contact with their families, first language, culture, nationality and religion. Almost all spent prolonged periods of time (in some cases years) in hospital when they no longer wanted or needed to be there. All those interviewed wanted to be discharged home far sooner. The generalizability of findings is unknown; however, important questions are posed in relation to nursing practice and research regarding the overall management of this group of young people. Issues include: respecting and upholding human rights; and challenging the boundaries of current nursing practice in order to move towards child-centred, appropriate and effective models of care. PMID- 10840256 TI - 'You feel helpless, that's exactly it': parents' and young people's control beliefs about bed-wetting and the implications for practice. AB - 'You feel helpless, that's exactly it': parents' and young people's control beliefs about bed-wetting and the implications for practice Young people wet the bed when they fail to wake up to a full bladder. The causes of bed-wetting are far from certain and this uncertainty is reflected in the diversity of treatments on offer and the lack of any guarantee that treatment will work in a particular case. Most young people are sad and ashamed about the bed-wetting and want it to stop, but they vary widely in their belief in their own capacity to influence the situation, and in their optimism about what the future holds. The problem can persist into adolescence or even adulthood, with far reaching social and emotional consequences, both for the young people and their families. In an ethnographic study involving 19 families 'perceived helplessness' emerged as a key issue permeating the whole system and often activating a downward spiral, leading to abdication of effort and responsibility by the young people themselves, by their parents and sometimes by health care professionals. Informed by the insights gained from this study and an extensive review of the literature on perceived control the Family Perspectives on Bed Wetting Questionnaire has been developed to explore family members' feelings, degree of concern and dimensions of perceived control relating to: effort, ability, luck, important others and the unknown. This questionnaire was used as a basis for structured interviews with family members in a longitudinal survey, involving 40 families attending one of nine community-based, nurse-led enuresis clinics in Greater Glasgow. It was found that only 38% of the young people were entering into treatment with the belief that they had the ability to be dry at night. This became self-fulfilling, with only 33% achieving initial success of 14 consecutive dry nights in a 16-week period (chi-squared test, P=0.029). Seventy per cent of young people felt that luck was important, while most parents felt that luck had little part to play. Implications for practice include the need to assess the families' readiness to engage in treatment, to create opportunities for effective control and to encourage realistic expectations. PMID- 10840257 TI - Researching children: issues arising from a phenomenological study with children who have diabetes mellitus. AB - In Britain the incidence of diabetes mellitus in children is increasing. These children are health care consumers and will continue to be throughout their lives. It is important that their views are sought regarding their health care condition. A phenomenological study was undertaken with six children aged 7-12 years regarding their experience of living with diabetes. This research highlighted several issues regarding the conduct of research with children. Access to children required negotiation with the ethics committee and the parents. It was also important to seek the consent of the children via the signing of a consent form, as well as an on-going process throughout the conduct of the research. In addition, appropriate communication methods had to be utilized, acknowledging the age and abilities of individual children. The use of both verbal and non-verbal communication skills were important, but the use of non-verbal skills appeared to be the most significant. Researchers should not be bound by the supposed abilities of children and should take care not to underestimate the awareness and maturity that some children possess when addressing issues of concern to themselves. When given the opportunity, children were keen to express their views and were very articulate in doing so. There is much scope for future research studies seeking children's perceptions about their health care condition. PMID- 10840258 TI - An exploration of the epistemological intricacies of using qualitative data to develop a quantitative measure of user views of health care. AB - Nurse researchers are increasingly combining qualitative and quantitative methods in order to understand more fully the world of research subjects. Qualitative data are often used to explore the subjective meanings behind survey responses and to develop quantitative measures and scales. Insights from qualitative data help researchers to design instruments which are more sensitive to respondents' meanings and interpretations. The aim of this paper is to highlight the epistemological and methodological complexities involved in this enterprise through drawing on our own experience of developing an instrument to examine person-centredness in health care from a qualitative study of dissatisfaction. The intricacies of this project relate to: epistemological continuity and inconsistency; research roles; reflexivity; confirmation; and completeness. Through discussing the literature around integrating methods, we suggest that researchers could be assisted in their attempts to develop conceptually sound quantitative measures by extending the concept of reflexivity (used in qualitative research) to the quantitative components of mixed method studies. This would aid conceptual clarity by making explicit the social, cultural, and political construction of knowledge, and would also encourage researchers to reflect upon the ethical and political consequences of their research. PMID- 10840259 TI - The use of simulation and post-simulation interview to examine the knowledge involved in community nursing assessment practice. AB - This paper describes the development of an innovative research approach which used the complementary methods of simulation and post-simulation interview to examine the knowledge-base involved in community nursing assessment practice in the United Kingdom. The study commenced in 1994 and the main phase of data gathering took place over a 3-week period in 1995. Having outlined the study's aim, context and theoretical background, this paper focuses on the two main methods of data-gathering used. Detailed description of the simulation method and the post-simulation interview and the rationales for their use are followed by critical discussion which identifies their particular strengths and weaknesses. Threats to validity are also considered. It is argued that the combined use of a simulated assessment and a post-simulation structured interview has great potential as a means of exploring the knowledge involved in community nursing assessment practice. PMID- 10840260 TI - Videotaped recording as a method of participant observation in psychiatric nursing research. AB - This paper describes videotaped recording as a data collection method when conducting participant observation in a psychiatric nursing study. The videotaped episodes were part of the daily life of psychiatric nursing in a hospital environment. The advantages and limitations of using videotaped recording in nursing research will be discussed. This paper is based on two studies. The data consisted of 21 videotaped episodes of nursing report sessions or interdisciplinary team meetings in the psychiatric clinic of a university hospital. The participants consisted of patients, their significant others, nurses, doctors, social workers and physiotherapists. All videotaped material was transcribed verbatim. An essential advantage of videotaping is that most potentially useful interaction and behaviour can be captured. The advantage in terms of the credibility of videotaping was that the investigator was able to review the same videotaped situations again and again. Videotaped material is rich and provides several possibilities for analysing the data. In these studies data and source triangulation enabled the researchers to reduce personal influence on the results. The investigator must also be aware of the limitations concerning this method. The most essential limitations are mechanical problems and the influence of videotaping on behaviour. Careful ethical considerations are important concerning personal privacy, informed consent and respect for the self determination of psychiatric patients. PMID- 10840261 TI - Is the complexity of care a paradox? AB - This paper looks at the phenomenon known as care and the medium through which it is expressed - caring. It explores some of the meanings of these terms but focuses particularly on nursing care. Superficially, nurses and society have a broad understanding of what 'care' means but common usage of the word belies its complexity. When examined alongside the writings of scholars the inconsistent nature of care and caring emerges. We reflect on the difficulties this presents for both the nurse and the cared for when, on the one hand care is promoted as the essence of nursing, while on the other there is no acceptable definition of care on which to base this claim. Encompassed within our discussion is the underlying theme that although care is an appropriate ideal for nursing it does not capture all of the day-to-day realities and hence it is not an overriding ideal. Care in nursing in this sense is contradictory and we describe it as paradoxical. PMID- 10840262 TI - Contributions of general internal medicine teaching units: a national survey. AB - OBJECTIVE: To identify and describe general internal medicine teaching units and their educational activities. DESIGN: A cross-sectional mailed survey of heads of general internal medicine teaching units affiliated with U.S. internal medicine training programs who responded between December 1996 and December 1997. MEASUREMENTS AND MAIN RESULTS: Responses were received from 249 (61%) of 409 eligible programs. Responding and nonresponding programs were similar in terms of university affiliation, geographic region, and size of residency program. Fifty percent of faculty received no funding from teaching units, 37% received full time (50% or more time), and 13% received part-time (under 50% time) funding from units. Only 23% of faculty were primarily located at universities or medical schools. The majority of faculty were classified as clinicians (15% or less time spent in teaching) or clinician-educators (more than 15% time spent in teaching), and few were clinician-researchers (30% or more time spent in research). Thirty six percent of faculty were internal medicine subspecialists. All units were involved in training internal medicine residents and medical students, and 21% trained fellows of various types. Half of the units had teaching clinics located in underserved areas, and one fourth had teaching clinics serving more than 50% managed care patients. Heads of teaching units reported that 54% of recent graduating residents chose careers in general internal medicine. CONCLUSIONS: General internal medicine teaching units surveyed contributed substantial faculty effort, much of it unfunded and located off-campus, to training medical students, residents, and fellows. A majority of their graduating residents chose generalist careers. PMID- 10840263 TI - Recent care of common mental disorders in the United States : prevalence and conformance with evidence-based recommendations. AB - OBJECTIVE: To relate the presence of recent mental disorders to use of mental health services. DESIGN: Cross-sectional survey. STUDY POPULATION: The study population was 3,032 respondents to the Midlife Development in the United States (MIDUS) survey, a nationally representative telephone-and-mail survey conducted in 1996. Twelve-month diagnoses according to the Diagnostic and Statistical Manual of Mental Disorders, Revised, Third Edition, of major depressive episode, panic disorder, generalized anxiety disorder, and serious mental illness were made using a structured assessment. Information was obtained on 12-month treatment for mental health problems in the general medical, mental health specialty, human services, and self-help sectors. Definitions of treatments consistent with evidence-based recommendations were developed using available practice guidelines. MEASUREMENTS AND MAIN RESULTS: Crude and adjusted likelihoods of receiving any mental health care and guideline-concordant care were measured. Although 53. 8% of respondents with at least one 12-month mental disorder received any mental health care in the previous year, only 14.3% received care that could be considered consistent with evidence-based treatment recommendations. Even among those with the most serious and impairing mental illness, only 25% received guideline-concordant treatment. Predictors of receiving guideline-concordant care included being white, female, severely ill, and having mental health insurance coverage. CONCLUSIONS: An epidemic of untreated and poorly treated mental disorders exists in the United States, especially among vulnerable groups such as African Americans and the underinsured. Cost-effective interventions are needed to improve both access to and quality of treatment. PMID- 10840264 TI - Randomized trial of case-finding for depression in elderly primary care patients. AB - OBJECTIVE: To determine the effect of case-finding for depression on frequency of depression diagnoses, prescriptions for antidepressant medications, prevalence of depression, and health care utilization during 2 years of follow-up in elderly primary care patients. DESIGN: Randomized controlled trial. SETTING: Thirteen primary care medical clinics at the Kaiser Permanente Medical Center, an HMO in Oakland, Calif, were randomly assigned to intervention conditions (7 clinics) or control conditions (6 clinics). PARTICIPANTS: A total of 2,346 patients aged 65 years or older who were attending appointments at these clinics and completed the 15-item Geriatric Depression Scale (GDS). GDS scores of 6 or more were considered suggestive of depression. INTERVENTIONS: Primary care physicians in the intervention clinics were notified of their patients' GDS scores. We suggested that participants with severe depressive symptoms (GDS score >/= 11) be referred to the Psychiatry Department and participants with mild to moderate depressive symptoms (GDS score of 6 -10) be evaluated and treated by the primary care physician. Intervention group participants with GDS scores suggestive of depression were also offered a series of organized educational group sessions on coping with depression led by a psychiatric nurse. Primary care physicians in the control clinics were not notified of their patients' GDS scores or advised of the availability of the patient education program (usual care). Participants were followed for 2 years. MEASUREMENTS AND MAIN RESULTS: Physician diagnosis of depression, prescriptions for antidepressant medications, prevalence of depression as measured by the GDS at 2-year follow-up, and health care utilization were determined. A total of 331 participants (14%) had GDS scores suggestive of depression (GDS >/= 6) at baseline, including 162 in the intervention group and 169 in the control group. During the 2-year follow-up period, 56 (35%) of the intervention participants and 58 (34%) of the control participants received a physician diagnosis of depression (odds ratio [OR], 1.0; 95% confidence interval [CI], 0.6 to 1.6; P =.96). Prescriptions for antidepressants were received by 59 (36%) of the intervention participants and 72 (43%) of the control participants (OR, 0.8; 95% CI, 0.5 to 1.2; P =.3). Two-year follow-up GDS scores were available for 206 participants (69% of survivors): at that time, 41 (42%) of the 97 intervention participants and 54 (50%) of the 109 control participants had GDS scores suggestive of depression (OR, 0.7; 95% CI, 0.4 to 1.3; P =.3). Comparing participants in the intervention and control groups, there were no significant differences in mean GDS change scores (-2.4 +/- SD 3.7 vs -2.1 SD +/- 3.6; P =.5) at the 2-year follow-up, nor were there significant differences in mean number of clinic visits (1.8 +/- SD 3.1 vs 1.6 +/ SD 2.8; P =.5) or mean number of hospitalizations (1.1 +/- SD 1.6 vs 1.0 +/- SD 1.4; P =.8) during the 2-year period. In participants with initial GDS scores > 11, there was a mean change in GDS score of -5.6 +/- SD 3.9 for intervention participants (n = 13) and -3.4 +/- SD 4.5 for control participants (n = 21). Adjusting for differences in baseline characteristics between groups did not affect results. CONCLUSIONS: We were unable to demonstrate any benefit from case finding for depression during 2 years of follow-up in elderly primary care patients. Studies are needed to determine whether case-finding combined with more intensive patient education and follow-up will improve outcomes of primary care patients with depression. PMID- 10840265 TI - Effects of depressive symptoms on health-related quality of life in asthma patients. AB - OBJECTIVE: To assess the effects of depressive symptoms on asthma patients' reports of functional status and health-related quality of life. DESIGN: Cross sectional study. SETTING: Primary care internal medicine practice at a tertiary care center in New York City. PATIENTS: We studied 230 outpatients between the ages of 18 and 62 years with moderate asthma. MEASUREMENTS AND MAIN RESULTS: Patients were interviewed in person in English or Spanish with two health-related quality-of-life measures, the disease-specific Asthma Quality of Life Questionnaire (AQLQ) (possible score range, 1 to 7; higher scores reflect better function) and the generic Medical Outcomes Study SF-36 (general population mean is 50 for both the Physical Component Summary [PCS] score and Mental Component Summary [MCS] score). Patients also completed a screen for depressive symptoms, the Geriatric Depression Scale (GDS), and a global question regarding current disease activity. Stepwise multivariate analyses were conducted with the AQLQ and SF-36 scores as the dependent variables and depressive symptoms, comorbidity, asthma, and demographic characteristics as independent variables. The mean age of patients was 41 +/- SD 11 years and 83% were women. The mean GDS score was 11 +/- SD 8 (possible range, 0 to 30; higher scores reflect more depressive symptoms), and a large percentage of patients, 45%, scored above the threshold considered positive for depression screening. Compared with patients with a negative screen for depressive symptoms, patients with a positive screen had worse composite AQLQ scores (3.9 +/- SD 1.3 vs 2.8 +/- SD 0.8, P <.0001) and worse PCS scores (40 +/- SD 11 vs 34 +/- SD 8, P <.0001) and worse MCS scores (48 +/- SD 11 vs 32 +/- SD 10, P <.0001) scores. In stepwise analyses, current asthma activity and GDS scores had the greatest effects on patient-reported health-related quality of life, accounting for 36% and 11% of the variance, respectively, for the composite AQLQ, and 11% and 38% of the variance, respectively, for the MCS in multivariate analyses. CONCLUSIONS: Nearly half of asthma patients in this study had a positive screen for depressive symptoms. Asthma patients with more depressive symptoms reported worse health-related quality of life than asthma patients with similar disease activity but fewer depressive symptoms. Given the new emphasis on functional status and health-related quality of life measured by disease-specific and general health scales, we conclude that psychological status indicators should also be considered when patient-derived measures are used to assess outcomes in asthma. PMID- 10840266 TI - Antidepressant treatment and health services utilization among HIV-infected medicaid patients diagnosed with depression. AB - OBJECTIVE: To characterize the prevalence and predictors of diagnosed depression among persons with HIV on Medicaid and antidepressant treatment among those diagnosed, and to compare utilization and costs between depressed HIV-infected individuals treated with and without antidepressant medications. DESIGN: Merged Medicaid and surveillance data were used to compare health services utilized by depressed individuals who were or were not treated with antidepressant medications, controlling for other characteristics. SETTING AND PARTICIPANTS: The study population comprised Medicaid recipients in New Jersey who were diagnosed with HIV or AIDS by March 1996 and received Medicaid services between 1991 and 1996. MEASUREMENTS AND MAIN RESULTS: Logistic regression and ordinary least squares regressions were employed. Women were more likely and African Americans were less likely to be diagnosed with depression. Women and drug users in treatment were more likely to receive antidepressant treatment. Depressed patients treated with antidepressants were more likely to receive antiretroviral treatment than those not treated with antidepressants. Monthly total expenditures were significantly lower for individuals diagnosed with depression and receiving antidepressant therapy than for those not treated with antidepressants. After controlling for socioeconomic and clinical characteristics, treatment with antidepressant medications was associated with a 24% reduction in monthly total health care costs. CONCLUSIONS: Depressed HIV-infected patients treated with antidepressants were more likely than untreated subjects to receive appropriate care for their HIV disease. Antidepressant therapy for treatment of depression is associated with a significantly lower monthly cost of medical care services. PMID- 10840267 TI - Differences in the quality of care for women with an abnormal mammogram or breast complaint. AB - OBJECTIVE: To examine factors associated with variation in the quality of care for women with 2 common breast problems: an abnormal mammogram or a clinical breast complaint. DESIGN: Cross-sectional patient survey and medical record review. SETTING: Ten general internal medicine practices in the Greater Boston area. PARTICIPANTS: Women who had an abnormal radiographic result from a screening mammogram or underwent mammography for a clinical breast complaint (N = 579). MEASUREMENTS AND MAIN RESULTS: Three measures of the quality of care were used: (1) whether or not a woman received an evaluation in compliance with a clinical guideline; (2) the number of days until the appropriate resolution of this episode of breast care if any; and (3) a woman's overall satisfaction with her care. Sixty-nine percent of women received care consistent with the guideline. After adjustment, women over 50 years (odds ratio [OR], 1.58; 95% [CI], 1.06 to 2.36) and those with an abnormal mammogram (compared with a clinical breast complaint: OR, 1.75; 95% CI, 1.16 to 2.64) were more likely to receive recommended care and had a shorter time to resolution of their breast problem. Women with a managed care plan were also more likely to receive care in compliance with the guideline (OR, 1.72; 95% CI, 1.12 to 2.64) and have a more timely resolution. There were no differences in satisfaction by age or type of breast problem, but women with a managed care plan were less likely to rate their care as excellent (43% vs 53%, P <.05). CONCLUSIONS: We found that a substantial proportion of women with a breast problem managed by generalists did not receive care consistent with a clinical guideline, particularly younger women with a clinical breast complaint and a normal or benign-appearing mammogram. PMID- 10840268 TI - Effects of primary care coordination on public hospital patients. AB - OBJECTIVE: To evaluate the effect of primary care coordination on utilization rates and satisfaction with care among public hospital patients. DESIGN: Prospective randomized gatekeeper intervention, with 1-year follow-up. SETTING: The Adult General Medical Clinic at San Francisco General Hospital, a university affiliated public hospital. PATIENTS: We studied 2,293 established patients of 28 primary care physicians. INTERVENTION: Patients were randomized based on their primary care physician's main clinic day. The 1,121 patients in the intervention group (Ambulatory Patient-Physician Relationship Organized to Achieve Coordinated Healthcare [APPROACH] group) required primary care physician approval to receive specialty and emergency department (ED) services; 1,172 patients in the control group did not. MEASUREMENTS AND MAIN RESULTS: Changes in outpatient, ED, and inpatient utilization were measured for APPROACH and control groups over the 1 year observation period, and the differences in the changes between groups were calculated to estimate the effect of the intervention. Acceptability of the gatekeeping model was determined via patient satisfaction surveys. RESULTS: Over the 1-year observation period, APPROACH patients decreased their specialty use by 0.57 visits per year more than control patients did ( P =.04; 95% confidence interval [CI] -1.05 to -0.01). While APPROACH patients increased their primary care use by 0.27 visits per year more than control patients, this difference was not statistically significant (P =.14; 95% CI, -0.11 to 0.66). Changes in low acuity ED care were similar between the two groups (0. 06 visits per year more in APPROACH group than control group, P =. 42; 95% CI, -0.09 to 0.22). APPROACH patients decreased yearly hospitalizations by 0.14 visits per year more than control patients (P =.02; 95% CI, -0.26 to -0.03). Changes in patient satisfaction with care, perceived access to specialists, and use of out-of network services between the 2 groups were similar. CONCLUSIONS: A primary care model of health delivery in a public hospital that utilized a gatekeeping strategy decreased outpatient specialty and hospitalization rates and was acceptable to patients. PMID- 10840270 TI - Managing patients with depression: is primary care up to the challenge? PMID- 10840269 TI - Telephone medicine for internists. AB - The role of the telephone in medical practice is important, but often problematic. Mistakes in telephone diagnosis and triage can have severe consequences. An effective office system can reduce liability risks, and in some cases telephone contact can substitute for office visits. Internists feel unprepared to provide telephone care. Therefore, residency education needs to focus on documentation, consultant availability, and performance feedback. Research should focus on improving outcomes, reimbursement issues, and technologic advances. This article describes internists' telephone interactions with ambulatory patients, preparation for telephone medicine, and aspects of office telephone systems and makes comparisons with other primary care fields. PMID- 10840271 TI - Who is doing the teaching? PMID- 10840272 TI - Searching for the best of primary care. PMID- 10840273 TI - Traumatic axonal injury: practical issues for diagnosis in medicolegal cases. AB - In the 25 years or so after the first clinicopathological descriptions of diffuse axonal injury (DAI), the criterion for diagnosing recent traumatic white matter damage was the identification of swollen axons ('bulbs') on routine or silver stains, in the appropriate clinical setting. In the last decade, however, experimental work has given us greater understanding of the cellular events initiated by trauma to axons, and this in turn has led to the adoption of immunocytochemical methods to detect markers of axonal damage in both routine and experimental work. These methods have shown that traumatic axonal injury (TAI) is much more common than previously realized, and that what was originally described as DAI occupies only the most severe end of a spectrum of diffuse trauma-induced brain injury. They have also revealed a whole field of previously unrecognized white matter pathology, in which axons are diffusely damaged by processes other than head injury; this in turn has led to some terminological confusion in the literature. Neuropathologists are often asked to assess head injuries in a forensic setting: the diagnostic challenge is to sort out whether the axonal damage detected in a brain is indeed traumatic, and if so, to decide what - if anything - can be inferred from it. The lack of correlation between well documented histories and neuropathological findings means that in the interpretation of assault cases at least, a diagnosis of 'TAI' or 'DAI' is likely to be of limited use for medicolegal purposes. PMID- 10840274 TI - Laminar specific loss of isocortical presenilin 1 immunoreactivity in Alzheimer's disease. Correlations with the amyloid load and the density of tau-positive neurofibrillary tangles. AB - Presenilin 1 has been shown to be mutated in a high proportion of cases of familial Alzheimer's disease. Immunoreactive epitopes of the protein have been found mainly in neurones devoid of neurofibrillary tangles - an observation that has led to the conclusion that presenilin 1 could have a protective role. In this study, the relationship between deposits of Abeta peptide (both the 40 and 42 isoforms), tau positive neurofibrillary tangles and presenilin 1-positive neuronal profiles were analysed in three cases of presenilin 1 mutation, four cases of sporadic Alzheimer's disease and five controls. Immunohistochemistry was performed in a sample from the supramarginal gyrus. The proportion of volume occupied by the Abeta1-40 and Abeta1-42 deposits (amyloid load) was evaluated by a point-counting technique. Tau-positive neurofibrillary tangles, and presenilin 1-positive neuronal profiles were directly counted. The location of the lesions in the thickness of the cortex was recorded. The density of PS1-positive neuronal profiles in Alzheimer's disease cases was lower than in the controls. The deficit was significant only in the upper layers of the cortex. The density of presenilin 1 neuronal profiles was negatively correlated with Abeta1-40 and Abeta1-42 loads, and with the density of tau-positive neurofibrillary tangles. Multivariate analysis showed that the Abeta1-42 load was the best determinant of the decrease in presenilin 1-positive neuronal profiles. Presenilin 1-positive neurones appear to be lost rather than protected in the course of Alzheimer disease. PMID- 10840275 TI - beta-amyloid (Abeta)42(43), abeta42, abeta40 and apoE immunostaining of plaques in fatal head injury. AB - beta-Amyloid (Abeta) deposits are found in the brains of approximately one-third of patients who die within days after a severe head injury; their presence correlating strongly with possession of an apolipoprotein E (apoE)-epsilon4 allele. The aim of the study was to investigate the relationship between Abeta42, Abeta40 and apoE immunostaining of Abeta plaques in the cerebral cortex and the relevance of apoE genotype in 23 fatally head-injured patients. These cases were known to have Abeta deposits from a previous study in which they were examined and semiquantified and related to apoE genotype. In the present study, the temporal cortex was probed using four different antibodies that recognize Abeta42(43), Abeta40 and an antibody to apoE. Abeta42(43)-positive plaques were observed in all of the 23 cases and Abeta40 immunoreactivity in only 11 of the 23 cases. In addition, semiquantitative analysis showed that relatively fewer plaques were detected with anti-Abeta40 than anti-Abeta42(43). ApoE immunoreactive plaques were identified in 18 of the 23 cases. The number of plaques stained for apoE was relatively less than for Abeta42(43) but greater than for Abeta40. Furthermore, the density of Abeta plaques detected using either Abeta42(43), Abeta40 or apoE antibodies was associated with possession of apoE epsilon4 in an allele dose-dependent manner. The results are consistent with Abeta42(43) as the initially deposited species in brain parenchyma and provide evidence that apoE is involved in the early stages of amyloid deposition. Further, the findings may be of relevance to the role of apoE genotype in influencing outcome after acute brain injury. PMID- 10840276 TI - Expression of the interferon-gamma-inducible chemokines IP-10 and Mig and their receptor, CXCR3, in multiple sclerosis lesions. AB - The recruitment of leucocytes to sites of inflammation is an important feature of multiple sclerosis (MS) pathology. Chemokines are involved in the activation and specific directional migration of monocytes and T-lymphocytes to sites of inflammation. Using immunocytochemistry, the expression of the alpha-chemokines, interferon (IFN)-gamma-inducible protein-10 (IP-10) and monokine induced by IFN gamma (Mig), and their receptor CXCR3 have been examined in post-mortem central nervous system (CNS) tissue from MS cases at different stages of lesion development. In actively demyelinating lesions both IP-10 and Mig protein were predominantly expressed by macrophages within the plaque and by reactive astrocytes in the surrounding parenchyma. CXCR3 was expressed by T cells and by astrocytes within the plaque. Interferon-gamma may stimulate glial cells to express IP-10 and Mig, which continue the local inflammatory response by selectively recruiting activated T-lymphocytes into the CNS. PMID- 10840277 TI - Dendritic and synaptic alterations of hippocampal pyramidal neurones in scrapie infected mice. AB - Neurone damage and eventual loss may underlie the clinical signs of disease in the transmissible spongiform encephalopathies (TSEs). Although neurone death appears to be through apoptosis, the trigger for this form of cell death in the TSEs is not known. Using two different murine scrapie models, hippocampal pyramidal cells were studied through microinjection of fluorescent dye, and synaptic integrity, using p38-immunoreactivity (p38-IR), both visualized using confocal laser scanning microscopy. Intradendritic distensions and dendritic spine loss were found to co-localize to areas of vacuolar and prion protein pathology in the hippocampus of mice infected with ME7 or 87 V scrapie. A significant reduction in p38-IR was found concomitantly in the hippocampus in ME7 scrapie mice. These results indicate that both pre- and post-synaptic sites are altered by scrapie infection; this would disrupt neuronal circuitry and may initiate apoptotic cell death, giving rise to the neurological disturbances manifested in clinical TSE cases. PMID- 10840279 TI - Tubulin immunoreactive neuronal intranuclear inclusions in the human brain. AB - Intranuclear filamentous and crystalline inclusion bodies have been described in the nuclei of a variety of cells in both normal and pathological states. The functional significance of these structures remains to be elucidated. Moreover, although the proteinaceous nature of these inclusions has been inferred in some histochemical studies, the identity of their constituent proteins remains to be determined. In the present study, immunohistochemistry was used to investigate the presence of intranuclear inclusions in neurones of the human brain which are intensely immunoreactive for the neuronal cytoskeletal protein class III beta tubulin. The ability to label these structures immunohistochemically was exploited to investigate the topographic pattern of distribution of these inclusions in the human brain. Intranuclear inclusions were rod-shaped, polygonal, or irregular in shape. They were present in neurones and ependymal cells. Intranuclear inclusion-bearing neurones were distributed in an anatomically heterogeneous pattern in the brain. Areas exhibiting relatively high densities of inclusions included the substantia inominata and anterior olfactory nucleus, dentate gyrus, substantia nigra, inferior olivary nucleus, and dentate nucleus of the cerebellum. In addition, intranuclear inclusions were prevalent in neurones in layers II, V, and VI of the cerebral cortex. They were particularly prevalent in the mesial basal temporal neocortex. The relationship of these structures to the intranuclear rods and sheets of the classical microscopists is uncertain. The demonstration that they are composed, at least in part, of tubulin, a major cytoskeletal protein, provides important clues regarding the mechanisms underlying their formation and provides a springboard for developing hypotheses regarding their functional significance. Furthermore, the ability to demonstrate these inclusions immunohistochemically provides an avenue for further studies directed at elucidating the potential involvement of these inclusions in various pathological settings. PMID- 10840278 TI - Motor neuronal death in sporadic amyotrophic lateral sclerosis (ALS) is not apoptotic. A comparative study of ALS and chronic aluminium chloride neurotoxicity in New Zealand white rabbits. AB - Whether diseased motor neurones in sporadic amyotrophic lateral sclerosis (ALS) die via apoptosis is unknown. Because this relates primarily to difficulties in utilizing post-mortem tissue from end-stage disease, motor neurone degeneration in ALS spinal cord was compared with that of a model of a chronic motor neurone degeneration. Degenerating motor neurones in ALS, identified by ubiquitin immunoreactivity, did not demonstrate the morphological characteristics of apoptosis and were not c-Jun immunoreactive or TUNEL positive. A temporal analysis of spinal motor neurone death in the chronic AlCl3 neurotoxicity model of motor neurone degeneration was also undertaken. AlCl3 was administered intracisternally every 4 weeks and, at intervals of 51, 107, 156 and 267 days, evidence of apoptosis was sought by morphology, TUNEL hybridization or DNA laddering. Double-labelling immunostudies were also performed with antibodies to either c-Jun, ubiquitin or high molecular weight neurofilament (NFH) with TUNEL hybridization. Although significant neurone loss was evident, apoptosis was not found. These studies demonstrate a lack of apoptosis in ALS spinal motor neurones and suggest that this observation does not relate to the utilization of post mortem tissue in which apoptotic neurones may have been lost. PMID- 10840280 TI - Cryopreservation of viable human glioblastoma xenografts. AB - Human tumour xenografts maintained in nude mice are a valuable research tool. The passaging and maintenance of human tumour xenografts in immune-deficient animals are expensive and labour-intensive. This study presents a protocol that permits long-term cryopreservation of viable glioblastoma xenograft tissue pieces in liquid nitrogen. Twenty different human glioblastoma xenografts that have been successfully transplanted and repeatedly passaged in nude mice were cryopreserved to validate the method. Different passages were cryopreserved for up to 40 months. On retransplantation of the tumours, all cases except one grew successfully. In order to ensure that the individual tumours did grow (not induced mouse tumours) restriction fragment length polymorphism analysis with variable number of tandem repeats (VNTR) probes was carried out. The method permits the long-term storage of viable, retransplantable glioblastoma xenografts. It minimizes animal use for the maintenance of xenografts and permits cryo-back-up of valuable tumours, thus markedly reducing the cost and increasing the accessibility of human tumour xenografts as a research tool in biology and genetics. PMID- 10840282 TI - Leadership, change, and free will. PMID- 10840281 TI - Correspondence: aesthesioneuroblastoma arising in pituitary gland. PMID- 10840283 TI - Identification of cardiovascular risk: use of a cardiovascular-specific genogram. AB - Cardiovascular disease (CVD) affects nearly 50 million Americans of all ages, races, and educational levels. Many of the risk factors for CVD are modifiable and public health nurses (PHNs) are in unique position to impact this major health problem because of their access to individuals, families, and groups. Addressing this major health problem requires early identification of those at risk for CVD. This article describes the implementation of a cardiovascular specific genogram (CVSG) which can be used to identify persons at risk for CVD. Rationale for the development of this disease-specific tool and suggestions for its clinical application are discussed. The genogram was distributed to the parents of 100 6th grade students. All of the respondents reported cardiovascular risk factors present in at least one of three generations examined. The risk factors in the two younger generations were at the primary and secondary levels of prevention and were modifiable with intervention. Only the older generation in this sample had tertiary level risk factors. The CVSG can easily be incorporated into all aspects of public health nursing practice, from assessment for case finding to planning and implementing disease management strategies. The CVSG can identify individuals, families, and groups at risk for CVD allowing the nurse to focus attention on those most at risk, and to implement proactive assessment, screening, and educational programs. PMID- 10840284 TI - Community based and community focused: nursing education in community health. AB - Nurses have always cared for individuals, families, groups, and communities in their practice. Recently there has been an increase in the focus on nurses working outside of the hospital, primarily in community-based settings that focus on individuals and families. There is also increasing emphasis on community focused nursing care with the community as the client. In some nursing programs, nurse educators have tried to adjust to this change by increasing the amount of time that nursing students spend in the community. The focus of this experience ranges from individuals to populations. This article describes a one-semester community health course in a baccalaureate nursing program that gives students both theoretical and practical knowledge in caring for individuals, families, groups, and communities. This allows for practice and understanding of both community-based and community-focused nursing care. PMID- 10840285 TI - Health promotion practices of the older adult. AB - The purposes of this study were to explore the overall primary and secondary health promotion activities of a group of older adults who resided in a life care community. This was a descriptive study, and included 194 out of 200 residents who participated in a single face-to-face interview. The majority of the participants were female (80%), white (98%), and cognitively intact (Mini Mental Status Exam [MMSE] scores of 27.0 +/- 4.1). Those in the oldest age group were less likely to monitor their cholesterol intake, have their stools checked for occult blood, or have a mammogram or Pap test over the previous 2 years. The most common reasons for not having these tests were advanced age, not being told to by their primary health care provider, and no interest in pursuing abnormal findings. Health care providers should consider an individualized approach to health promotion and disease prevention and help the older adult make his or her own decisions regarding participation in these activities. PMID- 10840286 TI - Impact of the Caring for Aging Relatives Group program: an evaluation. AB - Caregiver support programs have been developed with the goal of alleviating the stress associated with the demanding role of caring for an older person. The majority of these programs, however, have not been formally evaluated. This study is a formal program evaluation which assessed the impact of a caregiver support program on its participants. Both the intervention and matched comparison groups included a convenience sample of 23 female caregivers. A quasi-experimental pretest-posttest design was used to measure morale, social support, and information. Findings showed there was a positive relationship between morale and social support. In the comparison group only, there was a positive relationship between morale and information in both the pretest and posttest. Within the intervention group, there was a significant increase (t = 2.79, p = 0.01) in the information scores between the pretest and posttest. Caregivers in the intervention group reported that the Caring for Aging Relatives Group (CARG) provided them with social support. The results of this study partially supported the usefulness of a caregiver support program; that is, information was gained, morale was maintained, and caregivers perceived the support program as helpful. Future research is needed to address what factors lead caregivers to attend a support program and what type of social support they receive from attending such a program. PMID- 10840287 TI - Shopping center wheelchair accessibility: ongoing advocacy to implement the Americans with Disabilities Act of 1990. AB - Although the regulations of the Americans with Disabilities Act (ADA) of 1990 were phased in by 1992, monitoring and enforcement continue to be problematic. This study of three large shopping centers in the Southwest included one mall that was opened in the mid-1990s, and two malls that were constructed prior to the law (but have undergone recent renovations). Use of the ADA Accessibility Guidelines Checklist for Buildings and Facilities (Architectural and Transportation Barriers Compliance Board [ATBCB], 1992) generated data which were analyzed descriptively to determine the frequency and percent compliance in: parking lots, entrances, ramps, elevators, telephones, restrooms, food courts, and 12 specific store-types. No mall was fully compliant in any area, other than telephone specifications. In other areas, compliance ranged from 0% (ramp slopes in the newer mall) to many areas of 100% compliance (for example, outdoor curb ramps and food court seating spaces and aisles). The implications are that shoppers who are wheelchair mobile cannot count on complete compliance and cannot predict which physical architectural barriers they will find in shopping centers. PMID- 10840288 TI - Gender differences for the predictors of depression in young adults with genital herpes. AB - Genital herpes is a chronic, stigmatizing, sexually transmitted disease (STD), which is increasing despite efforts to control its spread. Depression is commonly reported among people diagnosed with genital herpes and differences in depression by gender have been reported. Therefore, the purpose of this study was to identify gender differences in the predictors of depression in young adults with genital herpes by secondary analyses of baseline data from a randomized clinical trial (RCT). For the RCT, young adults (193 females, 59 males) with genital herpes were recruited from newspaper advertisements. Participants completed questionnaires measuring illness burden, attitudes toward herpes, stress symptoms, mood states, depression, self-concealment, self-disclosure, substance use, and demographics. Univariate analyses and multiple regression techniques were used to identify variables predictive of depression in this sample. In women, increased anger, decreased vigor, increased confusion, a negative attitude toward herpes, self-concealment, and stress symptoms from genital herpes predicted more depression (R2 = 0.63). In men, increased depression was predicted by increased anger, a negative attitude toward herpes, and a decreased willingness to share personal information with a stranger (R2 = 0.51). Findings suggest that future psychoeducational interventions should address anger as a predictor of depression in this population. Gender-specific interventions need to be developed in order to assist young adults who are living with genital herpes. PMID- 10840289 TI - Performing a program evaluation in a family case management program: determining outcomes for low birthweight deliveries. AB - Program evaluations have become an integral part of public health practice. To illustrate the program evaluation process, an evaluation of clinical outcomes for clients of the Family Case Management (FCM) program administered by the nursing division of the DuPage County Health Department, Wheaton, Illinois, is presented. Birth outcomes for 1996 clients of the program who had a previous low-birthweight (LBW) infant and a subsequent birth using the prenatal services of the FCM program were analyzed. A purposive sample of clients was selected from the FCM prenatal program. Birth outcomes including infant gestational age and birthweight, maternal weight gain, and other key variables were compared to provision of health department services. Descriptive statistical methods were used to explore the relationship between birth outcomes and prenatal care between the first pregnancy and the second pregnancy. Analysis of the data showed improvements in maternal weight gain, infant birthweight and gestational age, prenatal care access, public health nurse (PHN) contacts, and physician visits. A positive association was shown between the early entry and increased use of prenatal services to improved birth outcomes in the subsequent pregnancy. PMID- 10840290 TI - A successful approach to immunizing men who have sex with men against hepatitis B. AB - This paper reports the success of outreach efforts to immunize adults at high risk of acquiring hepatitis B virus (HBV) infection. Local health department testers recruited clients when they sought anonymous HIV testing. Interested clients were serologically tested for HBV infection. Susceptibles were offered a free hepatitis B vaccine. Of 189 clients who were tested, 161 (86%) were susceptible to HBV infection and 140 (74%) were men who have sex with men (MSM). Of the susceptibles, 82 (51%) started the hepatitis B vaccine series. Sixty-five (80%) received three doses of hepatitis B vaccine and an additional 10 (12%) received two doses. Completing the hepatitis B vaccine series was associated with being a MSM (odds ratio [OR] 8.8, confidence interval [CI] 1.5-56.0) and with not being an injection drug user (IDU) (OR 0.2, CI 0.02-1.0). One way to provide hepatitis B vaccine to MSM is to attach an immunization program to a successful anonymous HIV testing program. The feasibility of implementing such programs would increase if public vaccine policymakers made more federally purchased hepatitis B vaccine available for adults. PMID- 10840291 TI - In harm's way: recognizing and addressing alcohol risk for rural disadvantaged pregnant mothers. AB - The following article describes the efforts of a group of rural Vermont maternal child health (MCH) nurses who addressed alcohol risk for the pregnant mothers they served through the Women, Infants, and Children (WIC) Supplemental Nutrition program. As front-line providers, MCH nurses are in a position to both improve the detection of problem drinking during pregnancy and to strengthen maternal alcohol prevention and intervention initiatives. Results showed that a more refined screening for alcohol provided an opportunity to promote harm reduction. PMID- 10840292 TI - Breastfeeding triplets: the at-home experience. AB - The unprecedented rise in higher-order multiple (HOM) births and the subsequent increase in parents who want to breastfeed their babies presents community health nurses (CHNs) with complex challenges. The extraordinary diversity of the triplet breastfeeding experience once all infants are settled at home is illustrated through findings from a survey of nine mothers of triplets. Parents revealed how they managed the feedings over the multiples' first year: type of feedings (breast, expressed breast milk [EBM], formula); scheduled and demand feedings; adequacy of milk supply; frequency and duration of feedings; consecutive and simultaneous feedings; nighttime with three; weaning; effects of breastfeeding on their bodies and well-being; challenges and stresses; and spousal, family, and health professional attitudes and support. A number of strategies that CHNs can utilize are suggested. These include working with individual families as well as forming partnerships with parents of multiples' support groups, multiple birth associations, interested health professionals, and the community sector. It is through these actions that the accessibility, coordination, and quality of health, multiple birth education, and social support services can be strengthened and healthy public policies implemented which address the unique and enormous demands experienced by HOMs' families. PMID- 10840293 TI - Economic evaluation of a metropolitan-wide, school-based hepatitis B vaccination program. AB - The results of an evaluation of a large metropolitan-wide, school-based hepatitis B vaccination program provide further evidence that such programs are effective and cost-beneficial. The percentage of 6th grade students fully immunized against hepatitis B rose from approximately 8% to 82% in the program. Administering the vaccine at school was $1.46 per dose less than traditional methods. Over $24 million of potential health care costs have been avoided through the program. PMID- 10840294 TI - Colonic neoplasia in patients with nonspecific GI symptoms. AB - BACKGROUND: The purpose of this study was to determine the prevalence rate of colonic polyps or masses 1 cm or greater in diameter in patients with nonspecific abdominal symptoms, from diverse practice settings, using a national endoscopic database. METHODS: Consecutive patients undergoing colonoscopy were included based on procedure indication. Endoscopic data were generated with a computer database at each practice site, transmitted to a central data bank and merged with data from multiple sites for analysis. Group 1 patients had nonspecific abdominal symptoms, which were defined as pain, constipation and diarrhea. Group 2 patients had a positive fecal occult blood test. Group 3 patients were asymptomatic, undergoing screening colonoscopy. Serious colon pathology was defined as a polyp or mass greater than 9 mm in size. RESULTS: Data were collected from 31 practice sites in 21 states during a period of 18 months. Of the 20,745 colonoscopy examinations, 9.2% were performed to evaluate patients with nonspecific abdominal symptoms, excluding other indications. Among patients with nonspecific symptoms 7.27% had polyp(s) 1 cm or greater in diameter compared with 17.05% of patients with positive fecal occult blood test (odds ratio 2.12: CI [1.73, 2.60]; p < 0.001). Patients with nonspecific symptoms had similar rates of large polyps as asymptomatic patients (7.27% vs. 6.45%, p = 0.32). Multivariate analysis identified several independent variables including increasing age, male gender and practice site at a Veterans Affairs Medical Center. CONCLUSIONS: In diverse, practice-based settings, patients with nonspecific abdominal symptoms who are referred for colonoscopy do not have a higher risk of serious colonic pathology than asymptomatic patients. PMID- 10840295 TI - Endoscopic sclerotherapy plus propranolol versus propranolol alone in the primary prevention of bleeding in high risk cirrhotic patients with esophageal varices: a prospective multicenter randomized trial. AB - BACKGROUND: Analysis of primary prevention studies of the use of beta-blockers has shown clear reductions in variceal bleeding in cirrhotic patients with varices. In contrast, the usefulness of prophylactic endoscopic sclerotherapy, alone or in combination with propranolol, in the management of these patients is still under investigation. The present study compared the efficacy of combined sclerotherapy and propranolol versus propranolol alone in the primary prevention of hemorrhage in cirrhotic patients with varices and high (greater than 18 mm Hg) intraesophageal variceal pressure. METHODS: Patients were randomly assigned to propranolol (42 patients) or to propranolol plus sclerotherapy (44 patients). The mean duration of follow-up was 26.8 +/- 7.7 and 24.6 +/- 9.8 months, respectively. RESULTS: During this period 23% of the patients in the combination group experienced at least 1 episode of bleeding due to varices or congestive gastropathy as compared with 14% in the propranolol group (not significant). Twenty-three patients (52%) in the combination group developed complications as compared with 8 (19%) in the propranolol group (p = 0.002). The mortality rate was similar in both groups (14% and 18%, respectively). The only independent factor predictive of survival was the level of serum albumin. CONCLUSIONS: Endoscopic sclerotherapy should not be used for the primary prevention of hemorrhage in cirrhotic patients at high risk of variceal bleeding who are undergoing treatment with propranolol. PMID- 10840296 TI - Ablation of Barrett's epithelium by endoscopic argon plasma coagulation in combination with high-dose omeprazole. AB - BACKGROUND: Barrett's esophagus is a premalignant condition induced by gastroesophageal reflux. The aim of this prospective study was to assess the efficacy of argon plasma coagulation in combination with high-dose omeprazole therapy to ablate nondysplastic Barrett's epithelium. METHODS: In 73 patients with histologically confirmed Barrett's epithelium, argon plasma coagulation was used in combination with maximal acid suppression (omeprazole 40 mg three times a day). Histologic and endoscopic changes were evaluated at 6- and 12-month intervals. RESULTS: In 69 of 70 patients (98.6%) complete squamous regeneration was achieved after a median of 2 argon plasma coagulation sessions (range 1 to 5). During a median follow-up of 12 months (range 2 to 51 months) there has been no relapse or evidence of the development of dysplasia under continuous acid suppression. Three patients (4.3%) developed a mild stricture of the distal esophagus that resolved after a single session of bougie dilation. CONCLUSIONS: In our experience, argon plasma coagulation in combination with high-dose omeprazole treatment is an effective and safe technique for complete ablation of nondysplastic Barrett's epithelium. Restoration of squamous mucosa after argon plasma coagulation appears to be long-lasting. PMID- 10840297 TI - Endoscopic biopsy requirements for post-treatment diagnosis of Helicobacter pylori. AB - BACKGROUND: Data on sensitivities of biopsy tests for Helicobacter pylori diagnosis after modern eradication therapy are limited. We assessed diagnostic yield of endoscopic biopsy tests before and after therapy in 2 U.S. multicenter double-blind trials of 10-day proton pump inhibitor-based triple therapy versus dual antibiotic therapy. METHODS: Three hundred one patients with duodenal ulcer and H pylori infection had endoscopy at baseline and at 8 weeks. Four antral and 3 body biopsies were taken at both endoscopies: 1 antral biopsy for a rapid urease test (CLOtest), 2 antral and 2 body biopsies for histologic examination (Genta stain), and 1 antral and 1 body biopsy for culture. RESULTS: The 2 same site biopsies (antral or body) for histologic examination were in agreement in 97% of cases before treatment and 100% after triple therapy. Histologic examination of antral biopsies without body biopsies missed H pylori infection in 2% of patients before treatment and 5% after triple therapy. Posttreatment sensitivities for triple therapy were significantly lower than pretreatment sensitivities for all tests (e. g., 18% decrease in sensitivity in antral histology, 22% decrease in antral culture); decreases in sensitivity were greater after triple therapy than after the less effective dual therapy. CLOtest plus histology had a post-treatment sensitivity of 96% in the triple therapy group. CONCLUSIONS: A single antral biopsy for histology provides excellent sensitivity for H pylori in untreated patients, but, after effective therapy, sensitivities of biopsy tests decrease. Use of more than one method of testing may increase diagnostic yield when assessing post-treatment H pylori status with endoscopy, whereas the addition of multiple biopsies for each type of test is of more limited value. PMID- 10840298 TI - Carbon urea breath test is not as accurate as endoscopy to detect Helicobacter pylori after gastrectomy. AB - BACKGROUND: This study was conducted to determine (1) whether Helicobacter pylori infection decreases in conjunction with time elapsed after gastrectomy and (2) the diagnostic efficacy of (13)C urea breath test (UBT) for H pylori in patients after gastrectomy. METHODS: From January 1997 to June 1998, 86 patients who had undergone gastrectomy and 180 patients with dyspepsia without gastrectomy were enrolled. A UBT for the analysis of excess (13)CO(2)/(12)CO(2) ratio (ECR) was obtained for each patient. Each patient also underwent endoscopy to obtain gastric biopsies for histology and H pylori culture. The presence of H pylori by either histology or culture served as the standard to test the efficacy of UBT. The 86 patients with a prior gastrectomy were categorized into 3 subgroups (I, less than 1 year; II, 1 to 3 years; III, greater than 3 years), according to the interval between surgery and UBT. The initial H pylori status of these 86 patients was determined by histologic evaluation of the resected stomach. RESULTS: At trial initiation, the postgastrectomy group had a lower H pylori infection rate (52.3%) as compared with the dyspeptic control group (80%). The initial H pylori status among subgroups I, II, and III was similar. There was a trend for the presence of H pylori in the stomach to decrease with increasing time elapsed after surgery (I to III: 68.8%, 48.3%, 36%, respectively; p < 0.05). The maximum UBT sensitivity and specificity achieved were 82.2% and 87.8% in the gastrectomy group and 97.2% and 96.3% in the dyspeptic group, with cutoff points of 2.5 and 4.0, respectively. CONCLUSION: The prevalence of H pylori diminishes with time elapsed after gastrectomy. UBT for detection of H pylori is more effective in patients without prior gastrectomy than in patients who have undergone gastrectomy and is less effective than endoscopy for patients who have had a gastrectomy. PMID- 10840299 TI - Colonoscopic polypectomy with cutting current: is it safe? AB - BACKGROUND: Coagulation and blended electrosurgical current are currently recommended for colonoscopic polypectomy, whereas pure cut current is believed to be associated with a higher risk of bleeding. However, the outcome of polypectomy performed with a cut current has not been evaluated in a large case series. Our objective was to study the incidence and nature of complications when polypectomy is performed with a pure cut current. METHODS: Among 9555 colonoscopic examinations, polypectomy cases were retrospectively reviewed for complications. The electrosurgical current applied was always the cutting waveform. RESULTS: Electrosurgical polypectomy using pure cut current was performed to remove 4735 lesions. Hemoclips were applied to the excision site after polypectomy to prevent bleeding in 12% of the cases. Hemorrhage occurred in 1.1% of the polypectomies (3.1% of patients). The incidence of bleeding with the different methods was snare polypectomy 0.9%, endoscopic mucosal resection 1.6%, "hot" biopsy 0.4%, and piecemeal polypectomy 7.3%. Bleeding was immediate in 66.1% of episodes and delayed in 33.9%. Patients with delayed postpolypectomy bleeding were significantly younger than those with immediate bleeding (50.5 and 64.7 years, respectively, p < 0.001). There was 1 case of transmural burn, but no perforations. CONCLUSION: Polypectomy can be performed with pure cut current with a bleeding rate comparable to that seen with the use of coagulation or blended current, provided that hemoclip placement can be used readily. Expertise in hemoclip placement is advisable if this method of polypectomy is to be used. PMID- 10840300 TI - Mistakes on EUS staging of colorectal carcinoma: error in interpretation or deception from innate pathologic features? AB - BACKGROUND: Because endoscopic ultrasound (EUS) accuracy for staging gastrointestinal tract tumors is limited by many factors, this study was designed to analyze potential sources of error in the EUS staging of colorectal carcinoma. METHODS: All patients referred for EUS evaluation of colorectal carcinoma were staged prospectively by one ultrasonographer and retrospectively by two others with EUS videotape recordings. Pathologic staging was done independently in a blinded fashion. Deceptive pathologic features were defined for T staging by presence of inflammation extending beyond tumor or microscopic spread without inflammation extending to a level consistent with the next stage, and for N staging by large (> or = 10 mm) benign lymph nodes or small (< 10 mm) malignant lymph nodes. RESULTS: Of 22 patients entered into the study, an inflammatory reaction around microscopic tumor spread thought to actually enhance detection by EUS was present in 57.1% of cases. Nine deceptive pathologic lesions were present in 36.4% (8 of 22) of patients (5 T stage, 4 N stage lesions). Of 40 T and N stage mistakes made by the three physicians, 45% were made in the presence and 55% in the absence of deceptive pathologic lesions. Accuracy increased significantly from the presence to absence of deceptive pathologic lesions, from 53.3% to 83.7% (p = 0.029) for T stage, and 8.3% to 73. 1% for N stage (p = 0.0001). Confidence of T staging correlated significantly with accuracy, increasing from 63.3% when unsure to 88. 2% with staging certainty (p = 0.017), an effect not seen for N staging. CONCLUSIONS: Inflammation and desmoplasia around colorectal carcinoma are often present, but may actually enhance EUS detection of microscopic tumor spread. Deceptive pathologic lesions are present in only one third of patients, but account for almost half (45%) of the errors in T and N staging by EUS. Diagnostic accuracy for EUS was increased with confidence in T stage assessment (but not N stage) and in the absence of deceptive pathologic lesions. Errors in interpretation still accounted for the majority of mistakes (55%) made in EUS staging of colorectal carcinoma. PMID- 10840301 TI - Postpolypectomy lower GI bleeding: descriptive analysis. AB - BACKGROUND: Postpolypectomy hemorrhage may warrant intensive care monitoring, transfusions, and surgery. We sought factors predicting significant bleeding requiring blood transfusion and the benefits of critical care monitoring. METHODS: Patients with postpolypectomy bleeding between April 1989 and November 1996 were identified from a comprehensive GI bleeding database. Data included age, gender, medical history, medications, polyp characteristics, and polypectomy technique. Outcomes assessed included bleeding cessation, transfusion requirements, recurrent bleeding, length of stay, and death. RESULTS: There were 83 patients with a median age of 73 years (range 18 to 88 years; 56 men, 27 women). Comorbid conditions were common (71.1% cardiovascular, 43.4% musculoskeletal, 14.5% hematologic, 6.0% renal). Within 3 days of presentation, 32.5% had taken aspirin, 10.8% nonsteroidal anti-inflammatory drugs, 12.0% warfarin, and 12.0% corticosteroids; and within 1 day, 10.8% intravenous heparin, 7.2% subcutaneous heparin, and 7.2% dipyridamole. Fifty-seven percent of patients were hemodynamically stable. Sessile cecal polyps greater than 2 cm in diameter bled more commonly. The median number of units transfused was equal between critical care and noncritical care patients. Using age in the logistic regression model, no other variable was predictive of transfusion. Eighty patients (96.4%) received endoscopic therapy, 1 required embolization and 2 hemicolectomy. There was no significant difference in outcomes for patients managed in an intensive care unit versus a general medical floor. CONCLUSIONS: Postpolypectomy bleeding appears to have a predictable presentation and outcome. Advanced age seems to be predictive of transfusion requirement. Patient monitoring in an intensive care setting is not absolutely necessary. PMID- 10840302 TI - Endoscopic piecemeal resection with submucosal saline injection of large sessile colorectal polyps. AB - BACKGROUND: Because endoscopic en bloc resection of large, sessile colorectal polyps is technically difficult, they are usually resected piecemeal. However, piecemeal resection makes it difficult to evaluate the completeness of the resection histopathologically. In this study the efficacy of endoscopic piecemeal resection of large, sessile colorectal polyps was investigated after follow-up greater than 1 year. METHODS: We removed 56 sessile colorectal polyps 2 cm or greater in diameter in 56 patients by using an endoscopic submucosal saline injection technique. Endoscopic examinations were repeated at 3, 6, and 12 months and longer after initial endoscopic resection. If no residual tumor was found endoscopically and histologically, the patient was considered to be "cured." RESULTS: Of the 56 polyps, 14 (25%) were resected en bloc, and 42 (75%) were resected piecemeal. Of the 42 patients treated with piecemeal resection, 23 (55%) required additional endoscopic or surgical interventions. In patients followed 1 year or longer after initial treatment, the cure rate by en bloc resection was 100% (14 of 14) and that by piecemeal resection was 83% (35 of 42). Arterial bleeding occurred in 4 patients (7%) during or after endoscopic resection. In 3 of them, bleeding was stopped by endoscopic clipping, but 1 patient required emergent laparotomy. CONCLUSIONS: Endoscopic piecemeal resection after submucosal saline injection with an intensive follow-up program is a safe and effective treatment for large, sessile colorectal polyps. PMID- 10840303 TI - Inhaled patient-administered nitrous oxide/oxygen mixture does not impair driving ability when used as analgesia during screening flexible sigmoidoscopy. AB - BACKGROUND: Entonox (nitrous oxide/oxygen mixture) has an analgesic effect, but its use is currently contraindicated in patients who plan to drive after its administration. We assessed the effect on driving ability of Entonox used as analgesia in patients undergoing screening flexible sigmoidoscopy. METHODS: Forty men and 40 women scheduled for screening flexible sigmoidoscopy who had not driven to the examination site were recruited together with a matched control subject from the same list. Before the examination complex motor skills were assessed in both groups using an adaptive tracking task, previously demonstrated to be the most sensitive test for detecting impairment of driving ability due to drugs. During the examination Entonox analgesia was available to the study group only, and after the screening examination complex motor function was retested on the same computer. RESULTS: Both groups improved their computer score on repeat testing, by 0.53 (range -1.9 to 4.6) in the study group and by 0.44 (range -0.6 to 2.5) in the control group. CONCLUSIONS: These data strongly support the view that Entonox analgesia does not impair driving ability. In the context of screening flexible sigmoidoscopy its use may lead to an increase in compliance. PMID- 10840304 TI - Prospective multicenter study comparing a standard reusable sphincterotome with a disposable triple-lumen sphincterotome. AB - BACKGROUND: Few data are available on the comparative performance of different types of sphincterotome. The aim of this study was to compare the efficacy of endoscopic sphincterotomy performed with either a reusable, single-lumen sphincterotome or a disposable triple-lumen instrument. METHODS: Seventy-seven consecutive adults requiring endoscopic sphincterotomy were prospectively and randomly assigned treatment with either a standard reusable single-lumen sphincterotome (group A, n = 38) or a disposable triple-lumen sphincterotome with a guidewire (group B, n = 39). The success rate, complications, and cost of the procedure per patient were compared. RESULTS: Deep cannulation was successful in 87% of cases in each group and sphincterotomy was achieved in 76% and 84% of cases in group A and B, respectively (NS). In the endoscopists' opinion the two instruments performed almost equally well. Twelve procedure-related complications occurred: 5 (all hemorrhages) in group A and 7 (1 hemorrhage, 4 cases of pancreatitis and 2 of cholangitis) in group B (NS). Eight reusable sphincterotomes were used with a cost of $61 per patient, compared with 39 disposable sphincterotomes with a cost of $241 per patient (p = 0.02). CONCLUSIONS: A standard reusable sphincterotome is satisfactory for most endoscopic sphincterotomies, and yields a substantial cost savings without compromising the success or safety of the procedure. PMID- 10840305 TI - Telomerase activity in pure pancreatic juice for the diagnosis of pancreatic cancer may be complementary to K-ras mutation. AB - BACKGROUND: The usefulness of K-ras mutation in pancreatic juice for the diagnosis of pancreatic cancer is questionable. Telomerase is positive in pancreatic cancer but rarely in benign pancreatic diseases. We conducted this study to determine the usefulness of K-ras mutation and telomerase activity in pancreatic juice for the diagnosis of pancreatic cancer. METHODS: Pancreatic juice collected during endoscopic retrograde cholangiopancreatography was examined in 31 patients: 12 with pancreatic cancer, 11 with chronic pancreatitis, and 8 control patients. The K-ras gene was detected by using the restriction fragment length polymorphism method. Telomerase activity was detected by using the telomeric repeat amplification protocol. RESULTS: K-ras mutation was positive in 75% (9 of 12) of pancreatic cancers and in 27% (3 of 11) of cases of chronic pancreatitis but in none of the control patients. Telomerase activity was detected in 92% (11 of 12) of pancreatic cancers and in 18% (2 of 11) of cases of chronic pancreatitis. The diagnostic value in pancreatic cancer was comparable between K-ras mutation and telomerase when evaluated separately. However, by combining these 2 methods, the specificity rose to 100%. CONCLUSIONS: For the diagnosis of pancreatic cancer, telomerase activity in pancreatic juice may possibly be complementary to K-ras mutation because it may decrease the rate of false-positive diagnosis. PMID- 10840306 TI - Reevaluation of duodenal endoscopic markers in the diagnosis of celiac disease. AB - BACKGROUND: Loss or reduction of duodenal folds, scalloping of Kerkring folds and a micronodular or mosaic duodenal mucosal pattern have been described in celiac disease (CD), endoscopic findings that are considered reliable in the diagnosis of this disorder. However, most data have been obtained in patients with suspected or certain disease. We assessed the accuracy of the above markers in diagnosing CD in patients with nonulcer dyspepsia. METHODS: In this prospective study, in 705 consecutive dyspeptic patients (284 men, 421 women, mean age 51 +/- SD 15.8 years) duodenal biopsies were obtained only in the presence of typical endoscopic markers, whereas in another 517 (207 men, 310 women, mean age 49.9 +/- SD 16 years) duodenal biopsies were done irrespective of macroscopic findings. CD was diagnosed histologically and on the basis of positive antiendomysium antibody. RESULTS: Endoscopic markers were found in 4 patients of the first group but CD was ruled out. In the second group 5 patients had an endoscopic pattern that was consistent and CD was diagnosed in 3, whereas 3 others with normal endoscopic findings were eventually diagnosed as having CD. Endoscopic markers had a sensitivity of 50% and a specificity of 99.6% (95% CI [11.8, 88.2 and 98.6, 99.9], respectively) with positive and negative predictive values of 60% and 99.4%, respectively. CONCLUSION: The accuracy of endoscopic markers in the diagnosis of CD must be reevaluated in relation to the characteristics of the population studied. PMID- 10840307 TI - All that scallops is not celiac disease. AB - BACKGROUND: Scalloping of duodenal folds as well as a mosaic mucosal pattern, decreased folds, and increased vascularity are markers of duodenal mucosal injury, the most common cause being celiac disease. We have recognized scalloping in patients with a variety of conditions other than celiac disease. METHODS: Clinical, endoscopic and histologic data were reviewed from selected patients with endoscopically visualized scalloped folds along with testing for endomysial antibodies. Biopsy specimens were examined histologically for villous:crypt ratio, intraepithelial lymphocytes, and inflammation. RESULTS: Thirteen patients with scalloped folds underwent endoscopy for the following reasons: family history of celiac disease and osteoporosis, gastrointestinal bleeding, dyspepsia (2), B(12)/ folate deficiency (4), and diarrhea (8). Histologic examination was abnormal in all but 1 patient. Villous atrophy or flattening as evidenced by reduced villous:crypt ratio was seen in 11 of 13 patients. Other abnormalities were edematous or broadened villi (10), intraepithelial lymphocytosis (7), and infiltration of lamina propria (6). An infectious organism was identified in 6 patients (46%). Celiac disease was excluded by the lack of specific biopsy findings combined with endomysial antibody testing. Final diagnoses were normal (1), eosinophilic enteritis (1), giardiasis (1), tropical sprue (4), human immunodeficiency virus-related diseases (6) including human immunodeficiency virus enteropathy (1). CONCLUSION: We conclude that scalloping is not specific for celiac disease but rather a predictor of mucosal disease as evidenced by villous atrophy, widening, and edema. PMID- 10840308 TI - Gastric cancer with calcification. PMID- 10840309 TI - Paracoccidioidomycosis masquerading as Crohn's disease. PMID- 10840310 TI - Aortoenteric fistula. PMID- 10840311 TI - Colonic intussusception. PMID- 10840312 TI - Wireless endoscopy. AB - BACKGROUND: Miniaturization of electronic components may allow the construction of new types of endoscopes that no longer require external wires, cables, or optical fibers. Our aim was to assess the feasibility of wireless endoscopy and to construct experimental prototypes using miniature charge-coupled device cameras, light sources, microwave transmitters, and batteries. METHODS: Feasibility, dimensions of miniature components, and power requirements were assessed. Prototypes were constructed and tested using cameras, transmitters, and halogen lamps powered by small batteries; 10.6 and 0.187 GHz transmitters were used to transmit the video signal. RESULTS: Moving television images were transmitted through models, post-mortem and live porcine stomachs, to the external receiver. Transmission of images through the abdomen was tested by placing the device in a microwave-impermeable box behind a volunteer's back and the receiver in front of his abdomen. In other experiments the endoscope was used inside the human mouth. The device was placed surgically in the stomachs of 150 kg pigs in vivo and good-quality color television image reception was achieved. CONCLUSIONS: These experiments demonstrate the feasibility of constructing a new type of endoscope that can transmit moving color television images from the GI tract without requiring fiberoptic or electrical cables. PMID- 10840313 TI - A novel approach to facilitate dilation of complex non-traversable esophageal strictures by efficient wire exchange using a stent pusher. AB - BACKGROUND: Dilation of non-traversable complex strictures is frequently difficult. For a tight stricture, over-the-wire dilation is difficult using a flexible biliary guidewire as opposed to a more rigid guidewire with a spring tip made for use with polyvinyl over-the-wire dilators (Savary). However, passage of the more rigid, spring-tipped wire is not always possible. A new method is discussed here whereby a biliary guidewire is exchanged consistently with a rigid, spring-tipped wire to facilitate dilation of complex strictures. METHODS: The wire exchange was carried out using a 10F stent pusher. The latter is readily available and semi-rigid, and its large lumen easily accommodates a Savary guidewire. Furthermore, because it can be passed through an endoscope, it can be passed without difficulty through an extremely tight stricture because the device does not buckle in the proximal esophagus or mouth. RESULTS: Dilation was successful in 6 patients using this technique with no complications. Four patients required placement of the stent pusher through a therapeutic endoscope. CONCLUSION: A method is described that facilitates dilation of complex, tight esophageal strictures by exchange of a flexible biliary guidewire with a more rigid Savary wire using a stent pusher. PMID- 10840314 TI - Endoscopic hemostasis using fibrin sealant for postsphincterotomy bleeding: report of two cases. PMID- 10840315 TI - Lymphangioma of the major duodenal papilla presenting as acute pancreatitis: treatment by endoscopic snare papillectomy. PMID- 10840316 TI - Closure of an esophageal perforation due to fish bone ingestion by endoscopic clip application. PMID- 10840317 TI - Esophageal metastasis of hepatocellular carcinoma. PMID- 10840318 TI - Synchronous multiple gastric cancers associated with familial gastric cancers. PMID- 10840319 TI - Stromal tumor as a pitfall in EUS-guided fine-needle aspiration cytology. PMID- 10840320 TI - Endoscopic resection of early-stage esophageal cancer accompanied by esophageal varices. PMID- 10840321 TI - Gastric mucosa-associated lymphoid tissue lymphoma with a focal high-grade component diagnosed by EUS and endoscopic mucosal resection for histologic evaluation. PMID- 10840322 TI - Combined laparoscopic and endoscopic retrieval of a migrated self-expanding metal stent. PMID- 10840323 TI - Intramural esophageal hematoma: a diagnostic dilemma. PMID- 10840324 TI - More on endoscopy databases. PMID- 10840325 TI - Excellence in endoscopy. PMID- 10840326 TI - Intubation and sedation in patients who have emergency upper GI endoscopy for GI bleeding. PMID- 10840327 TI - Laparoscopically assisted panenteroscopy for small bowel diseases: trans enterotomy versus peroral approach. PMID- 10840328 TI - A matter of semantics: hemorrhoids are a normal part of human anatomy and differ from hemorrhoidal disease. PMID- 10840329 TI - Unsedated peroral ultrathin videoendoscopy-assisted delivery of an esophageal manometry catheter. PMID- 10840330 TI - Response PMID- 10840331 TI - Treatment of Boerhaave's syndrome using the ultraflex self-expandable stent. PMID- 10840332 TI - Endoscopic cytology of biliary and pancreatic lesions. PMID- 10840333 TI - Response PMID- 10840334 TI - Guidelines for colorectal cancer screening and surveillance. PMID- 10840335 TI - Guidelines for training non-specialists in screening flexible sigmoidoscopy. PMID- 10840336 TI - Technology status evaluation report: ultrathin endoscopes esophagogastroduodenoscopy: March 2000. PMID- 10840337 TI - Technology status evaluation report: endoscopy simulators: May 1999. PMID- 10840338 TI - Technology status evaluation report: computerized endoscopic medical record systems: November 1999. PMID- 10840339 TI - 4th Decennial International Conference on nosocomial and healthcare-associated infections: a challenge for change. PMID- 10840340 TI - Hospital safety climate and its relationship with safe work practices and workplace exposure incidents. AB - BACKGROUND: In the industrial setting, employee perceptions regarding their organization's commitment to safety (i.e., safety climate) have been shown to be important correlates to both the adoption and maintenance of safe work practices and to workplace injury rates. However, safety climate measures specific to the hospital setting have rarely been evaluated. This study was designed to develop a short and effective tool to measure hospital safety climate with respect to institutional commitment to bloodborne pathogen risk management programs and to assess the relationship between hospital safety climate and (1) employee compliance with safe work practices and (2) incidents of workplace exposure to blood and other body fluids. METHODS: A questionnaire, which included 46 safety climate items, was developed and tested on a sample of 789 hospital-based health care workers at risk for bloodborne pathogen exposure incidents. RESULTS: A 20 item hospital safety climate scale that measures hospitals' commitment to bloodborne pathogen risk management programs was extracted through factor analysis from the 46 safety climate items. This new hospital safety climate scale subfactored into 6 different organizational dimensions: (1) senior management support for safety programs, (2) absence of workplace barriers to safe work practices, (3) cleanliness and orderliness of the work site, (4) minimal conflict and good communication among staff members, (5) frequent safety-related feedback/training by supervisors, and (6) availability of personal protective equipment and engineering controls. Of these, senior management support for safety programs, absence of workplace barriers to safe work practices, and cleanliness/orderliness of the work site were significantly related to compliance (P<.05). In addition, both senior management support for safety programs and frequent safety-related feedback/training were significantly related to workplace exposure incidents (P<.05). Thus the most significant finding in terms of enhancing compliance and reducing exposure incidents was the importance of the perception that senior management was supportive of the bloodborne pathogen safety program. CONCLUSIONS: Hospital safety climate with regards to bloodborne pathogens can be measured by using a short, 20-question scale that measures 6 separate dimensions. Whereas all 6 dimensions are essential elements of overall safety climate, 3 dimensions are significantly correlated with compliance, and 1 dimension (senior management support) is especially significant with regard to both compliance and exposure incidents. This short safety climate scale can be a useful tool for evaluating hospital employees' perceptions regarding their organization's bloodborne pathogens management program. In addition, because this scale measures specific dimensions of the safety climate, it can be used to target problem areas and guide the development of intervention strategies to reduce occupational exposure incidents to blood and other body fluids. PMID- 10840342 TI - Competency standards in the context of infection control. AB - BACKGROUND: To ensure quality patient care and enhance career development, competency levels of infection control professionals (ICPs) need to be identified and strengthened so that high standards of practice are established and maintained. OBJECTIVE: The purpose of this study was to apply a modified version of Benner's (1984) "Novice to Expert" model of skill acquisition to levels of competency and to seek to measure practices of the specialist practitioner in the context of infection control. METHOD: A self-administered questionnaire was developed and mailed to 464 members of the Infection Control Association, New South Wales (NSW) Inc, Australia. RESULTS: Seventeen percent of respondents reported full-time infection control responsibilities, 78% part time, and 5% unknown. The sample comprised 4 groups of ICPs. The largest groups were registered nurses, representing 37% of total respondents and "Other health care professionals" representing 35%. Forty-nine percent of the sample had completed a basic or advanced infection control certificate course, 21% had completed a bachelor of nursing or health science degree, and 21% had completed other studies not related to the specialty. Key findings of this study indicated that the clinical nurse consultant group rated their individual levels of skills and knowledge as proficient (competent). Although a small percentage from this group had completed higher level education, years of service within the specialty contributed to an increase in competent performance. This was also applicable to the other groups studied. CONCLUSION: The findings highlighted the need for a framework to be developed on which to build a model to measure and reflect progression of infection control competence at the beginner, advanced beginner, competent, and expert levels. Continuing education as a means of achieving competence needs to be further developed, maintained, and nurtured so that the ICP can acquire appropriate specialty knowledge and skills. PMID- 10840341 TI - Association between implementation of CDC recommendations and ventilator associated pneumonia at selected US hospitals. AB - BACKGROUND: To assess whether selected recommendations in the Centers for Disease Control and Prevention "Guideline for Prevention of Nosocomial Pneumonia" were being implemented and having an impact on the occurrence of ventilator-associated pneumonia (VAP) at US hospitals, we surveyed hospitals participating in the National Nosocomial Infections Surveillance (NNIS) system. METHODS: We mailed a questionnaire to the infection control practitioner of each NNIS hospital in 1995 and used data from the NNIS system to calculate annual rates of VAP. RESULTS: Of the 188 hospitals surveyed, 179 (95%) returned completed questionnaires. Of these, 175 (98%) had implemented the recommended change of mechanical-ventilator breathing circuits at 48-hour or greater intervals. Of 110 hospitals using the hygroscopic condenser-humidifiers or heat-moisture exchangers with ventilators, 102 (93%) changed the hygroscopic condenser-humidifiers or heat-moisture exchangers routinely, and of 98 hospitals using bubbling humidifiers, 96 (98%) used sterile water to fill these humidifiers. Other practices for which the guideline provides no recommendation and their frequency of use by NNIS hospitals include use of hygroscopic condenser-humidifiers or heat-moisture exchangers (110/179 [61%]) and use of bacterial filters in anesthesia machines (128/171 [61%]). There was a significant decrease in the VAP rate from 1987 to 1998. CONCLUSION: Most NNIS hospitals had implemented selected recommendations in the Centers for Disease Control and Prevention "Guideline for Prevention of Nosocomial Pneumonia" before the final publication of the revised guideline. Further studies are needed to assess the impact of these recommendations on the occurrence of VAP. PMID- 10840343 TI - Two-step tuberculin testing of passengers and crew on a commercial airplane. AB - OBJECTIVES: We investigated the risk of tuberculosis transmission from a person with highly infectious pulmonary tuberculosis to fellow passengers and crew members on a 14-hour commercial flight. The 2-step tuberculin testing was used to minimize the effects of the booster phenomenon. METHODS: Passengers and flight crew members identified from airline records were contacted by letter, telephone, or both to notify them of their potential exposure to Mycobacterium tuberculosis. The subjects were advised to undergo Mantoux tuberculin skin testing within the required time period to assess a conversion. In addition, information regarding tuberculosis history and other sources of potential exposure was solicited by means of a questionnaire. RESULTS: Of the 277 passengers and crew members on the aircraft, 225 (81.2%) responded. Of these, 173 (76.9%) had positive tuberculin results on the first test (induration > 10 mm). Thirteen subjects with negative results refused further testing; 11 (28%) of the remaining 39 exhibited the booster phenomenon on the second test. Subjects who exhibited the booster phenomenon were significantly more likely to have received previous BCG vaccination. Nine contacts with negative results on the initial test had positive results on a third test administered at 12 weeks after the flight exposure Of these, 6 contacts had previous BCG vaccination, old tuberculosis, or a family member with tuberculosis; the remaining 3 reported on other risk factors for positive reactions. None of these 3 contacts had sat in the same section of the plan as the index patient. CONCLUSIONS: The 2-step tuberculin testing procedure is an effective tool for minimization of the booster effect, thus allowing accurate monitoring of subsequent tuberculin conversion rates. Moreover, the clustering of tuberculin skin test conversions among passengers in this study demonstrates the possible risk of M tuberculosis transmission during air travel. PMID- 10840344 TI - Streptococcus pneumoniae: bacteremia in an era of penicillin resistance. AB - BACKGROUND: The proportion of penicillin-resistant Streptococcus pneumoniae isolates and associated risk factors varies by geographic area in the United States. We conducted a retrospective study to determine the extent of penicillin nonsusceptible S pneumoniae bacteremia and associated risk factors in a tertiary care medical center in San Diego. METHODS: Patients with S pneumoniae bacteremia at the University of California, San Diego Medical Center from September 15, 1991, through July 31, 1998, were identified by hospital-based computerized microbiology records. Hospital records included demographic information, patient data, and antibiotic prescription records for patients with bacteremia as a result of S pneumoniae. Univariate and multivariate analyses were used to determine risk factors for penicillin-nonsusceptible S pneumoniae bacteremia. RESULTS: Of 281 isolates of S pneumoniae identified, 192 (68%) were from hospitalized patients. After controlling for other factors, patients from 1 to 5 years of age (P = .01; odds ratio [OR] = 3.96; 95% CI, 1.50 to 10.44), 6 to 18 years of age (P =.04; OR = 6.42; 95% CI, 1.13 to 36.51), and HIV seropositive patients (P =.002; OR = 5.12; 95% CI, 1.83 to 14.32) were more likely to have penicillin-nonsusceptible S pneumoniae bacteremia. There was a significant increasing trend of penicillin-nonsusceptible S pneumoniae bacteremia from 14% in 1991 to 42% in 1998 (P = .001; OR = 1.42; 95% CI, 1.16 to 1.73); this included only 2 isolates that were highly resistant to penicillin. There was no increase in mortality in patients who had penicillin-nonsusceptible S pneumoniae bacteremia. CONCLUSION: With the increase in S pneumoniae resistance to penicillin, it is important to continue surveillance of infections caused by S pneumoniae. Hospital-based studies are useful for tracking epidemiologically important pathogens. PMID- 10840345 TI - Increasing antimicrobial resistance in gram-negative bacilli isolated from patients in intensive care units. AB - BACKGROUND: We investigated gram-negative bacilli from patients in intensive care units to determine whether antimicrobial resistance was increasing. METHODS: Minimal inhibitory concentrations were determined by broth microdilution on 334 gram-negative bacilli collected in 1990, 1995, and 1998. RESULTS: During the 3 study years, the types of gram-negative bacilli encountered in our intensive care units changed with proportional increases of Pseudomonas sp and decreases of inducible enterics. Dramatic increases in resistance for ceftazidime, cefotaxime, and piperacillin were paralleled between respiratory-tract isolates and inducible enterics. By 1998, ticarcillin was more active than piperacillin against most isolates except Escherichia coli and Klebsiella sp, and most isolates became more resistant to gentamicin and tobramycin. CONCLUSIONS: Continuous changes in the types of gram-negative bacilli and antimicrobial resistance complicate empirical selection of antimicrobials in the intensive care units. These complications will place more emphasis on communication and strategy formations among health care workers (nurses, physicians, laboratorians, and pharmacists) in an effort to treat infections in a timely and effective manner. PMID- 10840346 TI - Microbicidal activity of MDI-P against Candida albicans, Staphylococcus aureus, Pseudomonas aeruginosa, and Legionella pneumophila. AB - BACKGROUND: MDI-P (Medical Discoveries, Inc-Pharmaceutical, Layton, Utah) is a clear, colorless liquid generated by electrolysis of preservative-free and endotoxin-free, nonpyrogenic, sterile, injection saline (0.9% NaCl, wt/vol). It contains numerous highly reactive chlorine and oxygen species, including HOCl( 1,) OCl-(1), Cl(-1), Cl(2), O(2-)(1), and O(3). This report presents data on the in vitro microbicidal activity of MDI-P against 4 clinically relevant microbial pathogens that are often difficult to eradicate. METHODS: MDI-P was generated from injection saline by using a patented electrolysis instrument. It was then tested for microbicidal activity at concentrations ranging from 0.01% to 50% against Staphylococcus aureus, Pseudomonas aeruginosa, Legionella pneumophila, and Candida albicans (10(5) to 10(9) colony-forming units/mL). The effect of serum (50% and 90%) and pH on MDI-P activity were also tested. The morphologic effects of MDI-P on microbial cells were studied by light microscopy of cells stained by Gram's method and by transmission electron microscopy. Morbidity, mortality, and the effect of MDI-P on tissues were studied by using a mouse model. RESULTS: The microbicidal activity of MDI-P occurred within the first minute of exposure for all the organisms tested. When 50% MDI-P was tested against cell titers of 10(5) or 10(7) colony-forming units/mL, all test organisms were killed within 1 minute; at lower MDI-P concentrations, C albicans was the most sensitive organism, and L pneumophila was the most resistant. Even with beginning cell titers of 10(9) colony-forming units/mL, killing by 50% MDI-P was >99.9% for all test strains. Furthermore, at the same beginning cell titer, killing of C albicans by MDI-P diluted to 50% with normal human serum rather than injection saline was only slightly reduced. No acute morbidity, mortality, or tissue damage was detected in mice that were intravenously given 17 mL/kg of undiluted MDI-P. CONCLUSIONS: MDI-P is a very fast-acting, broad-spectrum microbicidal material. The lack of evidence for acute morbidity, mortality, or tissue injury, ease of preparation, and low cost suggest that it may be useful for various sterilization and disinfection applications. PMID- 10840347 TI - Enterobacter cloacae sepsis outbreak in a newborn unit caused by contaminated total parenteral nutrition solution. AB - OBJECTIVE: The study aimed to investigate an outbreak caused by Enterobacter cloacae in a neonate intensive care unit. DESIGN: A descriptive study of an outbreak of sepsis in high-risk neonates was used. SETTING: The study was set in a tertiary care university teaching hospital. PATIENTS: The patients were 11 neonates infected with Enterobacter cloacae whose symptoms and signs of sepsis developed during a 16-hour period. All but one neonate received parenteral nutrition. Isolates from blood cultures, in-use parenteral nutrition solutions, and control aliquots of parenteral nutrition solution were typed by pulsed-field gel electrophoresis. RESULTS: Enterobacter cloacae was found in the refrigerated aliquots of parenteral nutrition solution, in blood cultures from infected newborns, and from in-use parenteral nutrition solutions. All these strains of Enterobacter cloacae had the same antibiotic susceptibility pattern and the same genomic DNA profile. The strain isolated from the one patient who did not receive parenteral nutrition presented a different susceptibility profile and genotype. CONCLUSION: The source of the nosocomial sepsis was the parenteral nutrition solution in 10 neonates. This contamination apparently occurred during preparation of the parenteral solution. PMID- 10840348 TI - HIV and diarrhea in the era of HAART: 1998 New York State hospitalizations. AB - BACKGROUND: This study reflects an attempt to identify the causes of diarrheal illness in hospitalized HIV patients in light of therapeutic advancements in HIV management. METHODS: The study identifies the various etiologies associated with diarrhea among HIV patients hospitalized in New York State in 1998. Data for this study were extracted from the New York State Department of Health Statewide Planning and Research Cooperative System. Pathogens recognized to cause diarrhea in persons with HIV and general codes identifying diarrhea were examined by using the principal and all secondary diagnoses based on the International Classification of Diseases 9th Revision Clinical Modification codes. RESULTS: Based on the Statewide Planning and Research Cooperative System data set, more than 15,000 patients with HIV were hospitalized in 1998. Among the HIV patients hospitalized, 2.8% were admitted with a diarrheal diagnosis. The following diagnoses occurred the most frequently among HIV patients hospitalized with a diarrheal illness: Clostridium difficile (51.3%), other protozoal diseases (18.1%), and other organisms, not elsewhere specified (11.7%). CONCLUSIONS: In the era of highly active antiretroviral therapy, diarrhea is still an occurring symptom in HIV patients. Despite the relatively small percentage of hospitalizations attributed to diarrhea, clinicians must remember that even "mild" to "moderate" diarrhea can have a debilitating impact among persons with the symptom. PMID- 10840349 TI - Evaluation of a satellite education program on the prevention and control of antimicrobial resistance. PMID- 10840350 TI - Nasal colonization by methicillin-resistant coagulase-negative staphylococcus in community skilled nursing facility patients. AB - BACKGROUND: Methicillin-resistant coagulase-negative staphylococci (MRCNS) are increasing nosocomial pathogens in acute care hospital patients. However, there is little information on the epidemiology of MRCNS in skilled nursing facilities (SNFs). We report a pilot survey of the prevalence of MRCNS colonization in SNF patients. METHODS: Anterior nasal swabs were plated on oxacillin salt screening agar for selection of MRCNS. Suspected MRCNS were confirmed by coagulase and catalase tests and standard disc-diffusion antimicrobial susceptibility tests. RESULTS: The overall prevalence of MRCNS was 40% for in-house continuing SNF patients, 49% for newly admitted patients, and 60% for SNF nursing personnel. The prevalence was 13% in a "control" group of nonmedical personnel. Forty-six percent of MRCNS were resistant to ciprofloxacin. The frequency of colonization with MRCNS increased over time. After an average 17 months of facility stay, 32% of noncarriers acquired MRCNS. High frequency of colonization was associated with greater disability. CONCLUSION: Colonization with MRCNS is common among SNF patients, who can serve as a reservoir for transfer of such strains to acute care hospitals. Careful infection control practice, including judicious use of antibiotics with frequent handwashing, will remain critical policies for limiting spread of such strains. PMID- 10840351 TI - Hand hygiene rates unaffected by installation of dispensers of a rapidly acting hand antiseptic. AB - OBJECTIVE: The objective of the study was to improve health care workers' compliance with hand hygiene after patient contact by use of an alcohol-based hand antiseptic. DESIGN AND METHODS: Six commercially available alcohol-based hand antiseptics were evaluated. The one most pleasing to the evaluators' hands was selected for the study. Baseline handwashing rates were assessed on 2 medical wards. Alcohol dispensers were mounted by every door on the 2 wards. An educational campaign was conducted with 4 weekly visits to these floors to remind and reinstruct staff about the use of the alcohol dispensers and to address questions. After 2 months handwashing rates were reassessed. SETTING: The study was set in a university hospital. RESULTS: The baseline handwashing rate was 60% (76/126). Physicians were most compliant (83%), followed by nurses (60%), technologists (56%), and housekeepers (36%). Two months later overall hand hygiene rates had decreased to 52% (P = .26). Nurses were most compliant (67%), followed by technologists (57%), physicians (29%), and housekeepers (25%). Physician compliance was associated with compliance by attending physicians whose example was usually followed by all other physicians on rounds. CONCLUSIONS: A brief educational campaign and installation of dispensers containing a rapidly acting hand hygiene product near hospital rooms did not affect hand hygiene compliance. The behavior of attending physicians was predictive of handwashing rates for all others in the attending's retinue. Compliance with handwashing after half of all patient contacts was a result of perfect compliance by some and total noncompliance by others being observed. PMID- 10840352 TI - Blood substitutes in surgery. PMID- 10840353 TI - Clinical significance of microscopic tumor venous invasion in patients with resectable hepatocellular carcinoma. AB - BACKGROUND: Tumor venous invasion in patients with resectable hepatocellular carcinoma (HCC) is frequent and can be macroscopic and microscopic or microscopic alone. Although macroscopic invasion is a well-established prognostic indicator, the clinical significance of microscopic invasion remains unclear. METHODS: There were 322 patients enrolled who had undergone curative resection for HCC. The clinicopathologic factors and prognostic significance associated with macroscopic and microscopic venous invasion were analyzed. RESULTS: Macroscopic invasion was observed in 50 patients (15.5%) and microscopic invasion in 190 (59.0%). The larger the tumor, the more the incidence of venous invasion. There were 140 patients with microscopic invasion only (Group 1). Patients with macroscopic invasion (Group 2, n = 50) also had microscopic invasion. Compared with patients without venous invasion (Group 3, n = 132), Group 1 had a higher alpha fetoprotein level, a larger tumor size, and more tumors without encapsulation. For group 1, the 1-, 3-, and 5-year disease-free survival rates were 65.6%, 41.6%, and 30.8%, respectively. The 1-, 3-, and 5-year overall survival rates were 87. 8%, 60.0%, and 52.7%, respectively. The survival rates of group 1 were lower than those of group 3 and higher than those of group 2 (P <.05). Multivariate analysis indicated that microscopic and macroscopic venous invasion, surgical margin, indocyanine-green retention, and tumor size and number were significant predictors of postresectional survival. CONCLUSIONS: In HCC patients, microscopic venous invasion is frequent and related independently to postresectional outcome. PMID- 10840354 TI - Hepatobiliary scintigraphy is superior to abdominal ultrasonography in suspected acute cholecystitis. AB - BACKGROUND: Hepatobiliary scintigraphy is a very accurate test in the diagnosis of acute cholecystitis. However, ultrasonography is extensively used for the diagnosis of this disease. In this study, we directly compare the diagnostic accuracy of these techniques for acute cholecystitis. MATERIALS AND METHODS: The diagnostic accuracy of scintigraphy and ultrasonography was evaluated in 107 consecutive patients with suspected acute cholecystitis who underwent both imaging modalities within one day. The incremental diagnostic value of each modality was determined. RESULTS: The sensitivity, specificity, positive and negative predictive values, and accuracy for the diagnosis of acute cholecystitis in the entire cohort were superior for scintigraphy compared with ultrasonography. The accuracy was 92% for scintigraphy and 77% for ultrasonography. Similarly, if only surgically treated patients were considered, the accuracy of scintigraphy was 91% versus 61% for ultrasonography. The diagnostic value of scintigraphy for the entire cohort was significantly superior to ultrasonography (global, chi(2) = 58.1 vs 9.7, respectively); the addition of the information derived from the latter did not further improve the diagnostic value of scintigraphy (global, chi(2) = 58.2). CONCLUSIONS: Hepatobiliary scintigraphy has superior diagnostic accuracy for acute cholecystitis compared with ultrasonography. The addition of ultrasonography does not further improve the diagnostic accuracy of scintigraphy alone. PMID- 10840355 TI - Liver function and encephalopathy after partial vs direct side-to-side portacaval shunt: a prospective randomized clinical trial. AB - BACKGROUND: The aim of this study was to determine, in a prospective randomized clinical trial, whether the partial portacaval shunt offers any advantage in terms of liver function and encephalopathy rate when compared with direct side-to side direct portacaval shunt. METHODS: Forty-six "good risk" patients with cirrhosis and with documented variceal hemorrhage were randomly assigned to either a partial shunt procedure (achieved by 10-mm diameter interposition portacaval H-graft) or direct small-diameter side-to-side portacaval anastomosis. RESULTS: Operative mortality was zero in both groups. During the follow-up period, encephalopathy developed in 3 patients in the partial shunt group and 9 in the direct shunt group (P =.04). Kaplan-Meier analysis demonstrated that encephalopathy-free survival was significantly longer in the partial shunt group (P =.025). Direct shunt patients had significant hepatic functional deterioration postoperatively compared with the partial shunt group. CONCLUSIONS: The partial portacaval shunt effectively controls variceal hemorrhage. Compared with direct side-to-side portacaval shunt, partial shunt preserves long-term hepatic function and minimizes postoperative encephalopathy. We conclude that the partial portacaval shunt is the preferred approach over direct shunts for patients with cirrhosis and with variceal bleeding. PMID- 10840356 TI - Polypoid lesions of the gallbladder: report of 100 cases with special reference to operative indications. AB - BACKGROUND: The nature of polypoid lesions of the gallbladder is difficult to define before operation, and surgical indications still remain controversial. The aim of this study was to identify characteristics of each type of polypoid lesion of the gallbladder and indications for surgery. METHODS: Clinical data were retrospectively correlated with the histopathologic characteristics of polypoid lesions in 100 patients who had cholecystectomy. RESULTS: There were 74 benign polypoid lesions, including 39 cholesterol polyps, 20 adenomas, and 15 with adenomyomatous hyperplasia and 26 malignant polypoid lesions. Twenty-seven percent of patients with benign polyps and 73 percent of patients with malignant polyps were over 60 years of age. Polypoid lesions of the gallbladder were diagnosed by preoperative ultrasonography in only 36 patients (36%). All types of polypoid lesions of the gallbladder, whether benign or malignant, were frequently solitary, and gallstones coexisted in the majority of patients with all polypoid lesions of the gallbladder except cholesterol polyps. The lesions were > 10 mm in 88% of the malignant polyps and in only 15% of the benign polyps. CONCLUSIONS: The risk factors for malignancy were the age of the patient ( >60 years), the coexistence of gallstones, and the size of the polypoid lesions (>10 mm in diameter). In asymptomatic patients, cholecystectomy can be justified if there are risk factors for malignancy. PMID- 10840357 TI - Local ampullary resection with careful intraoperative frozen section evaluation for presumed benign ampullary neoplasms. AB - BACKGROUND: Frozen section evaluation has been reported to be inaccurate in detecting foci of adenocarcinoma within adenomas of the ampulla of Vater, leading many authors to advocate pancreaticoduodenectomy as the method of treatment for these neoplasms. The authors hypothesized that (1) ampullary resection is less morbid than pancreaticoduodenectomy, and (2) frozen section evaluation following ampullary resection is accurate and allows for a selective application of pancreaticoduodenectomy to those with carcinoma or benign lesions too large to be locally resected. METHODS: A retrospective review of a single-surgeon experience was conducted. Thirty-eight patients who underwent ampullary resection and pancreaticoduodenectomy (39 procedures) for benign and malignant ampullary neoplasms were identified. Our technique of step-frozen section analysis is described. RESULTS: Twenty-one ampullary resections were performed for preoperative diagnoses of benign (16) and malignant (5) ampullary neoplasms. Frozen section evaluation accurately predicted the final histology in all patients undergoing ampullary resection. Ampullary resection (vs pancreaticoduodenectomy) was associated with a statistically lower operative time (169 minutes vs 268 minutes), estimated blood loss (192 mL vs 727 mL), mean length of stay (10 days vs 25 days), and overall morbidity (29% vs 78%). CONCLUSIONS: Frozen section evaluation of ampullary neoplasms is accurate. Because ampullary resection is less morbid than pancreaticoduodenectomy and frozen section evaluation is accurate, ampullary resection with frozen section evaluation is our current approach to the treatment of small benign ampullary neoplasms. PMID- 10840358 TI - Surgical risks and outcome of pancreas retransplants. AB - BACKGROUND: The increased popularity of pancreas transplants has led to an increased number of potential candidates for retransplants after the initial graft has been lost to technical failure or rejection. We studied a group of recipients who underwent pancreas transplants at a single center to determine whether retransplant recipients were at higher risk of complications. METHODS: Between June 1, 1994, and Dec 31, 1997, a total of 213 pancreas transplants were performed at the University of Minnesota. Of these, 187 were primary transplants and 26 were retransplants. Demographically, the two groups were not significantly different. We analyzed and compared the two groups with respect to incidence of surgical complications, graft survival rates, and patient survival rates. RESULTS: Surgical complications such as bleeding and vascular thrombosis were slightly more common after retransplants, but this trend did not quite reach statistical significance. Infectious complications and leaks were equivalent between the two groups. The incidence of acute rejection was higher after retransplants (P =.02). At 3 years posttransplant, patient survival was no different between the two groups, but pancreas graft survival was lower after retransplants (P =.08). The incidence of early graft loss (by 6 months posttransplant) was significantly higher in retransplant recipients (27% vs 14%, P =.04). CONCLUSIONS: Pancreas retransplants can be performed with a minimal increase in surgical complications. However, graft survival after retransplants is slightly inferior to that after primary transplants, probably for both immunologic and nonimmunologic reasons. Retransplants can be offered to suitable candidates, but they may require more aggressive monitoring for rejection. PMID- 10840359 TI - Macrophage-derived transforming growth factor-beta1 induces hepatocellular injury via apoptosis in rat severe acute pancreatitis. AB - BACKGROUND: The mechanism of acute pancreatitis-induced hepatocellular injury is unclear. We have observed hepatocyte apoptosis in rat acute necrotizing pancreatitis. These studies were designed to determine the mediator(s) responsible for hepatocyte apoptosis and to clarify the significance of macrophages as its source. METHODS: A rat sodium deoxycholate-induced pancreatitis model was used. Immunohistochemical studies for apoptosis-inducing mediators on hepatocytes were examined in the liver and on the peritoneal macrophages. The levels of transforming growth factor-beta1 (TGF-beta1) were also evaluated quantitatively with an enzyme-linked immunosorbent assay. Induction of apoptosis on the hepatocytes was evaluated by in situ nick-end labeling and tissue DNA fragmentation enzyme-linked immunosorbent assay. Finally, the effects of TGF-beta1 neutralization and macrophage depletion were examined. RESULTS: In the liver and the peritoneal macrophages, strong expression of TGF-beta1 was detected early in the course of pancreatitis. In sodium deoxycholate-induced pancreatitis, the levels of TGF-beta1 were also elevated in the plasma (9.2 +/- 0.8 ng/mL), in the pancreatitis-associated ascitic fluid (11.5 +/- 0.6 ng/mL), and in the liver homogenate (2.8 +/- 0.3 ng/g of liver tissue). Moreover, the amount of fragmented DNA of the liver with pancreatitis was 290% +/- 20% of that with a sham operation and serum alanine aminotransferase levels elevated to 248.2 +/- 67.0 IU/L. TGF-beta1 neutralization partly blocked the positive labeling on the nuclei of the hepatocytes, the elevation of the amounts of fragmented DNA (205% +/- 10% of sham operation), and the serum alanine aminotransferase level (144.2 +/- 14.9 IU/L). On the other hand, the macrophage depletion caused a marked decrease in the TGF-beta1 protein level in the plasma (4.8 +/- 1.2 ng/mL) or in the pancreatitis-associated ascitic fluid (8.0 +/- 1.0 ng/mL). Moreover, the macrophage depletion completely inhibited the elevation of the TGF-beta1 protein level in the liver homogenate (1.5 +/- 0.4 ng/g of liver tissue), and thereafter decreased the amounts of the positive labeling on the nuclei of the hepatocytes and decreased the amount of fragmented DNA (120% +/- 18% of sham operation) and the serum alanine aminotransferase elevation (119.2 +/- 24.2 IU/L). CONCLUSIONS: In a model of sodium deoxycholate-induced pancreatitis, macrophages are responsible for pancreatitis-induced hepatocellular injury by means of apoptosis, and macrophage-derived TGF-beta1 is one of the major factors inducing the hepatocyte apoptosis. PMID- 10840360 TI - Role of hyaluronan in acute pancreatitis. AB - BACKGROUND: The connective tissue component hyaluronan is accumulated locally in the damaged tissue during various inflammatory conditions. Owing to the strong water-binding capacity of this glycosaminoglycan, increased tissue content of hyaluronan is paralleled by the development of interstitial edema. The aim with the current experiment was to investigate whether hyaluronan is accumulated in acute pancreatitis and if increased levels of hyaluronan can be correlated to the inflammation of the pancreatic tissue. METHODS: Acute pancreatitis was induced in Sprague-Dawley rats by the administration of supramaximal doses of the cholecystokinin analogue caerulein. The animals were followed for 5 hours (n = 4), 24 hours (n = 6), or 48 hours (n = 5), and the pancreata were then investigated for hyaluronan and water content, hyaluronan distribution, general morphology and the presence of CD44-positive cells, macrophages, and T lymphocytes. RESULTS: Hyaluronan accumulated in the edematous interstitium during acute pancreatitis. Twenty-four hours after the induction of pancreatitis, the hyaluronan content of the pancreata had increased by more than 100%. Simultaneously, CD44-positive cells infiltrated the tissue. However, no correlation between hyaluronan and water was seen at any time point. CONCLUSIONS: This study shows that acute pancreatitis is associated with a strong but transient increase in interstitial hyaluronan and an infiltration of CD44 positive cells located mainly in the same region as the accumulated hyaluronan. PMID- 10840361 TI - Invited commentary: does hyaluronan play a role in acute pancreatitis? PMID- 10840362 TI - A novel hydroxyl radical scavenger, nicaraven, protects the liver from warm ischemia and reperfusion injury. AB - BACKGROUND: Reactive oxygen species have been considered to be involved in liver injury at the procurement, preservation, and transplantation from donors without beating hearts. A novel hydroxyl radical scavenger, nicaraven with hydrophilic and lipophilic properties, infiltrates both intracellular and extracellular spaces where it effectively scavenges reactive oxygen species. Protection by nicaraven against ischemia and reperfusion damage of the brain, heart, and kidneys has been shown. The effect of this agent on the liver remains unclear. METHODS: Two-hour total hepatic vascular exclusion was used. Eighteen beagle dogs were randomly assigned to 2 groups: 12 animals were not treated (group I) and 6 were treated with nicaraven (group II). Nicaraven was administered intravenously (2mg/kg/min) for 60 minutes before ischemia and for 3 hours, starting 30 minutes before reperfusion. RESULTS: Two-week survival rates were 25% in group I and 100% in group II (P <.01). Nicaraven inhibited lipid peroxidation in the liver, improved hepatic and systemic hemodynamics and energy metabolism, and suppressed liver enzyme release, endothelin-1 elevation in hepatic venous blood, histologic damage, and neutrophil infiltration into the liver. CONCLUSIONS: Nicaraven exerted hepatic protection against warm ischemia and reperfusion injury. This may indicate nicaraven as a potential candidate to attenuate liver injury from warm ischemia and preservation in transplantation from donors without beating hearts. PMID- 10840363 TI - Adenovirus-mediated insulin gene transfer improves nutritional and post hepatectomized conditions in diabetic rats. AB - BACKGROUND: Impaired nutritional conditions in patients with diabetes are significant risk factors after major abdominal surgery. We constructed recombinant adenovirus vector carrying the human insulin gene (AxCAIns) for in vivo insulin gene transfer to improve metabolic impairments after a major operation in patients with diabetes. We tested the effects of AxCAIns on nutritional and post-hepatectomized conditions in rats with diabetes treated with streptozotocin (STZ). METHODS: AxCAIns was injected into the spleen in diabetic rats treated with STZ. Blood levels of glucose, total protein, albumin, and C peptide of human proinsulin were measured and the expression of transferred human insulin gene was analyzed in various organs. Diabetic rats underwent 70% partial hepatectomy with or without AxCAIns injection, and post-hepatectomized conditions were analyzed. RESULTS: STZ-induced hyperglycemia was reduced by AxCAIns injection. Decreased serum levels of total protein and albumin in diabetic rats were significantly restored to normal levels by AxCAIns injection, and human C peptide was detected in the sera of AxCAIns-treated rats. Human preproinsulin messenger RNA, which represented the expression of transferred insulin gene, was detected in the liver and the spleen, but not in other organs. Serum albumin levels, remnant liver weight, and ratios of postoperative body weight to preoperative body weight were significantly increased by AxCAIns in hepatectomized diabetic rats. CONCLUSIONS: AxCAIns injection to the spleen efficiently transferred the human insulin gene mainly into the hepatocytes and produced enough human proinsulin to improve nutritional impairments and post hepatectomized conditions in diabetic rats. Insulin gene transfer with intrasplenic injection of AxCAIns may be available to improve metabolic impairment after major abdominal surgery in patients with diabetes. PMID- 10840364 TI - Gender does not impact infrainguinal vein bypass graft outcome. AB - BACKGROUND: The percentage of women requiring infrainguinal bypass graft operations continues to increase, whereas the effect of gender on postoperative outcome remains unclear. The purpose of this study was to assess the influence of gender on patient selection and outcome in patients requiring infrainguinal vein bypass grafting procedures. METHODS: This retrospective study reviewed 217 infrainguinal vein bypass grafts performed over an 8-year period. Medical records and patient interviews were used to determine study measures and outcomes. Gender and multiple covariables affecting patient survival were analyzed; postoperative complications and graft patencies were examined. Bivariate and life-table analyses were conducted, followed by multivariate analysis with the Cox proportional hazards model. RESULTS: No statistical differences existed between men and women for age, diabetes, cardiac disease, tobacco use, hypertension, stroke, renal disease, or prior contralateral bypass or major amputation. Women were more likely to be black (P =.014) and have a spliced vein graft (P =.035). No differences were noted between the 2 groups for 30-day morbidity rates-except women had more incisional complications (P =.01)-or for survival (P =.45), primary-patency (P =.57), secondary-patency (P =. 79), or limb-salvage rates (P =.40). Multivariate analysis showed that gender had no role in affecting survival rates. CONCLUSIONS: Gender does not affect graft patency, limb salvage, or survival rates. There should be no introduction of a gender bias into management of infrainguinal occlusive disease. PMID- 10840365 TI - Insulin and glucocorticoid-dependent suppression of the IGF-I system in diabetic wounds. AB - BACKGROUND: Growth-promoting polypeptides, including insulin-like growth factor-I (IGF-I), orchestrate different biochemical events that culminate in the restoration of functional integrity of wounded skin. The nonhealing cutaneous wound is a well-documented phenomenon in experimental and clinical diabetes. Accordingly, we undertook this study to ascertain whether diabetes impairs the healing process by suppressing the wound microenvironmental IGF-I system (eg, IGF I; IGF-I receptor [IGF-I R]; and IGF-I binding protein [IGF-BP(3)]). METHODS: The induction of diabetes was achieved by the intravenous injection of streptozotocin at a dose of 55 mg/kg. Subcutaneously implanted polyvinyl alcohol sponge and stainless steel mesh chamber models were used to study wound healing. Nondiabetic and diabetic animals received, respectively, subcutaneous 30-day time-release pellets of glucocorticoid (200 mg) and mifepristone (RU-486, 25 mg). Corresponding control animals received placebo pellets. Polyvinyl alcohol sponge and wound fluid expression of the IGF-I system were evaluated by using ligand blotting, radioimmunoassay, and reverse transcriptase polymerase chain reaction based techniques. RESULTS: Polyvinyl alcohol sponge contents of messenger RNA (mRNA) transcripts encoding for IGF-I, IGF-I R, and IGF-BP(3) were reduced in diabetic and glucocorticoid-treated control animals. A similar pattern of changes in protein levels of IGF-I and IGF-BP(3) occurred in wound fluid collected from these animals. Partial normalization of the associated hyperglycemic and hypercortisolemic states of diabetes with insulin (hyperglycemia) and the glucocorticoid receptor blocker RU-486 (hypercortisolemia) ameliorated the diabetes-related decrease in the IGF-I system during wound healing. CONCLUSIONS: The current data, together with data garnered from the literature, support the concept that the state of hypercortisolemia in diabetes mellitus impairs the healing process, at least in part, by suppressing the wound microenvironmental IGF-I system. Confirmation regarding this premise awaits further investigation. PMID- 10840366 TI - Vitronectin deficiency is associated with increased wound fibrinolysis and decreased microvascular angiogenesis in mice. AB - BACKGROUND: Vitronectin has several putative functions including regulating hemostasis, cell adhesion, and cell migration. However, the targeted deletion of vitronectin in mice results in normal development and normal coagulation parameters. To determine whether vitronectin may be necessary for nondevelopmental processes, we examined the response to tissue injury in vitronectin-null mice. METHODS: We examined wound healing in control and vitronectin-null mice by healing rate, zymography, reverse zymography, and Western blots. RESULTS: We found that dermal wound healing was slightly delayed in mice lacking vitronectin. More importantly, we found extensive areas of delayed hemorrhage near the sprouting tips of microvessels between days 7 and 14, which temporally coincided with increased urokinase-type plasminogen activator and tissue-type plasminogen activator activity by zymography. Though Western blots confirmed the presence of plasminogen activator inhibitor-1 protein throughout wound repair and reverse zymograms showed decreased plasminogen activator inhibitor-1 activity between days 7 and 14. CONCLUSIONS: Loss of vitronectin in mice was associated with changes in the fibrinolytic balance, and this may have led to focal sites of delayed hemorrhage. The mechanism that resulted in decreased angiogenesis and the formation of larger blood vessels in response to tissue injury remains unknown. This study suggests that vitronectin may have several distinct functions that are not required for normal development but are manifested in response to tissue injury. PMID- 10840367 TI - Surgical ethics in a day's work: a personal vignette. PMID- 10840368 TI - Mucinous cystadenocarcinoma of the mesentery. PMID- 10840369 TI - Mycetoma - an unusual site. PMID- 10840370 TI - A replaced common hepatic artery running through the pancreatic parenchyma. PMID- 10840371 TI - Psychiatric presentation of carotid stenosis. PMID- 10840373 TI - Minimal pulmonary embolism detected after abdominal insufflation for laparoscopic cholecystectomy. PMID- 10840372 TI - Thrombin injection for treating a subclavian artery pseudoaneurysm. PMID- 10840374 TI - Gene therapy trials face further scrutiny. PMID- 10840375 TI - Why genetic testing of adoptive children should be restricted. PMID- 10840376 TI - Better access to clinical trials information. PMID- 10840377 TI - Medicines from safer GMOs? PMID- 10840378 TI - New approaches to treating haemophilia. PMID- 10840379 TI - Rapid update PMID- 10840380 TI - HLA-B27 misfolding: a solution to the spondyloarthropathy conundrum? AB - Compelling evidence indicates that HLA-B27 is directly involved in the etiopathogenesis of the spondyloarthropathies (SpAs). Several hypotheses based on its native antigenic structure, the peptides it presents and mimicry with bacterial epitopes, have been proposed. However, these potential mechanisms remain largely unsupported by human studies and transgenic animal models. Recent work demonstrating that HLA-B27 misfolds offers a novel alternative hypothesis. Here, we review this new information on the folding and assembly of HLA-B27, and discuss consequences of misfolding that could be relevant to the pathogenesis of SpAs. PMID- 10840381 TI - A cardiovascular EST repertoire: progress and promise for understanding cardiovascular disease. AB - The application of expressed sequence tag (EST) technology has proven to be an effective tool for gene discovery and the generation of gene expression profiles. The generation of an EST resource for the cardiovascular system has revealed significant insights into the changes in gene expression that guide heart development and disease. Furthermore, an important genetic resource has been developed for cardiovascular biology that is valuable for data mining and disease gene discovery. PMID- 10840383 TI - Towards a molecular therapy for glycogen storage disease type II (Pompe disease). AB - Glycogen storage disease type II (GSD-II), also known as Pompe disease, is a fatal genetic muscle disorder caused by a deficiency of acid alpha-glucosidase, a glycogen-degrading lysosomal enzyme. Currently, there is no treatment for this fatal disorder. However, several lines of research suggest the possibility of future treatment. Enzyme replacement strategies hold the greatest hope for patients currently affected by GSD-II, but future strategies could include in vivo or ex vivo gene therapy approaches and/or mesenchymal stem cell or bone marrow transplantation approaches. Each of the approaches might eventually be combined to further improve the overall clinical efficacy of any one treatment regimen. The lessons learned from GSD-II research will also benefit a great number of individuals affected by other genetic disorders. PMID- 10840382 TI - Genetic clues to the biological basis of autism. AB - Autism, the prototypical pervasive developmental disorder, is characterized by impaired communication and social interaction, and by repetitive interests and behaviours. The core disorder probably affects around 5:10 000 individuals, of whom some three-quarters are male. Onset is in the first three years of life, and the disorder is associated with lifelong disabilities. Because of the clear evidence that idiopathic autism has a strong genetic basis, many groups are undertaking whole genome screens to identify susceptibility loci. We review the first results, and briefly consider the implications of molecular genetic findings for future research, diagnosis and management. PMID- 10840385 TI - Small is beautiful. PMID- 10840384 TI - Neurofibromatosis type II: mouse models reveal broad roles in tumorigenesis and metastasis. PMID- 10840386 TI - Glioblastoma: encouraging the body to fight back. PMID- 10840388 TI - Bilateral brain stimulation for advanced Parkinson's disease. PMID- 10840389 TI - Therapeutic potential for orally administered type 1 interferons. AB - Interferons (IFNs), with annual global sales valued at more than US$4 billion, have therapeutic value in the treatment of viral, neoplastic and autoimmune diseases. Parenteral administration by high-dose injection can, however, cause serious side effects. Significant improvement in the therapeutic index of IFNs could be achieved with oral administration. Using this route, dose-related side effects are not seen, and efficacy is maintained in both animal studies and human clinical trials. Oral IFN administration appears to mimic a natural innate immune response. As such, it may represent an alternative delivery strategy to make better use of these critical cytokines. PMID- 10840387 TI - Augmenting cancer vaccine strategies with dendritic cell boosts. PMID- 10840390 TI - Swellable matrices for controlled drug delivery: gel-layer behaviour, mechanisms and optimal performance. AB - The majority of oral drug delivery systems (DDS) are matrix-based. Swellable matrices are monolithic systems prepared by compression of a powdered mixture of a hydrophilic polymer and a drug. Their success is linked to the established tabletting technology of manufacturing. Swellable matrix DDS must be differentiated from true swelling-controlled delivery systems. This review focuses on hydrophilic swellable matrix tablets as controlled DDS. Gel-layer behaviour, front movement and release are described to show the dependence of the release kinetics on the swelling behaviour of the system. In vivo behaviour of matrix systems is also considered. PMID- 10840391 TI - Liposomal delivery of camptothecins. AB - The use of liposomal drug delivery systems to improve the therapeutic index of pharmaceutical agents is exemplified by camptothecin-based drugs. This highly active class of anticancer agents possesses a unique mechanism of action with some inherent shortcomings, which might have been solved by liposomal formulation. Recent studies have revealed the protective action, increased tumor delivery and prolonged plasma exposure of these liposomal formulated drugs. These advances in pharmaceutical development could increase the levels of activity of these agents, as well as increase their clinical utility as new emerging anticancer therapies. PMID- 10840392 TI - Monitor: progress and profiles. PMID- 10840393 TI - Comparison of intrinsic optical signals associated with low Mg2+-and 4 aminopyridine-induced seizure-like events reveals characteristic features in adult rat limbic system. AB - PURPOSE: To analyze the intrinsic optical signal change associated with seizure like events in two frequently used in vitro models-the low-Mg2+ and the 4 aminopyridine (4-AP) models-and to monitor regions of onset and spread patterns of these discharges by using imaging of intrinsic optical signals (IOS). METHODS: Combined hippocampal-entorhinal-cortex slices of adult rats were exposed to two different treatments: lowering extracellular Mg2+ concentrations or application of 100 microM 4-AP. The electrographic features of the discharges were monitored using extracellular microelectrodes. Optical imaging was achieved by infrared transillumination of the slice and analysis of changes in light transmission using a subtraction approach. The electrographic features were compared with the optical changes. Regions of onset and spread patterns were analyzed in relevant anatomic regions of the slice. RESULTS: Both lowering extracellular Mg2+ concentrations and application of 4-AP induced seizure-like events. The relative duration of the intrinsic optical signal change associated with seizure-like events in the low-Mg2+ model was significantly longer compared with that seen with those occurring in the 4-AP model, although duration of field potentials did not differ significantly in the two models. Seizure-like events of the low-Mg2+ model originated predominantly in the entorhinal cortex, with subsequent propagation toward the subiculum and neocortical structures. In contrast, no consistent region of onset or spread patterns were seen in the 4-AP model, indicating that the seizure initiation is not confined to a particular region in this model. CONCLUSIONS: We conclude that different forms of spontaneous epileptiform activity are associated with characteristic optical signal changes and that optical imaging represents an excellent method to assess regions of seizure onset and spread patterns. PMID- 10840394 TI - Magnetic resonance imaging follow-up of progressive hippocampal changes in a mouse model of mesial temporal lobe epilepsy. AB - PURPOSE: Hippocampal sclerosis (HS) is the most frequent lesion found in mesial temporal lobe epilepsy (mTLE). MR imaging is considered to be the most sensitive and specific method to detect HS. Despite extensive studies performed on humans and except in a recent study, the morphologic pattern of HS is usually analyzed when the disease has already fully developed, thus not allowing any insight into the mapping of the progressive morphologic changes inducing the development of mTLE. We have recently characterized a model of mTLE that reproduces the unilateral pattern of HS, induced by intrahippocampal injection of low doses of kainate (KA) in mice. METHODS: In this study, we monitored the temporal evolution of the development of HS in this model of mTLE by using T2-weighted sequence, T2 relaxation time measurements, and T1-weighted spin-echo technique after injection of gadolinium, from 1 h to 120 days after KA injection. RESULTS: HS induced by intrahippocampal KA injection occurred in two phases. First, we observed a transient hyperintense T2-weighted signal in the cortex above the injected hippocampus, most likely indicative of vasogenic edema partly due to the neurotoxic effect of KA. The concomitant increase in the T2 signal in the injected hippocampus and ipsilateral amygdala likely reflects the phase of cytotoxic edema occurring probably in relation to the excitotoxic consequences of both KA and seizure activity. Second, from 15 days on, a persistent unilateral increased T2 signal was detected in the hippocampus, which most probably reflects gliosis. CONCLUSIONS: Our findings indicate that longitudinal follow-up would permit a better understanding of the mechanisms underlying the constitution of HS in humans and eventually development of prevention strategies. PMID- 10840395 TI - Changes in benzodiazepine binding in a subkindling situation. AB - PURPOSE: A low dose of the benzodiazepine receptor inverse agonist methyl beta carboline-3-carboxylate (beta-CCM) (1 mg/kg) was used to assess [3H]-flumazenil binding in a subkindling situation in Swiss mice. METHODS: The brains were removed, and benzodiazepine receptor binding was studied every second day over 14 days of administration. RESULTS: With each successive trial, Bmax values showed a steady and significant decrease, whereas Kd values showed a steady and significant increase. Behavioral data showed that at this low dose, actual kindling (seizuring) was not reached at the behavioral level. CONCLUSIONS: The findings suggest that decreased gamma-aminobutyric acid (GABA) inhibition may occur even if behavioral effects of kindling are not observed. PMID- 10840396 TI - Effect of long-term vigabatrin administration on the immature rat brain. AB - PURPOSE: To determine whether the neuropathologic changes produced by vigabatrin (VGB; gamma-vinyl GABA) administration in the developing rat brain are reversible. METHODS: We injected rats daily with VGB (25-40 mg/kg/day, s.c.) from age 12 days for 2 weeks followed by 2 weeks of a drug-free period. Behavioral testing, magnetic resonance (MR) imaging, biochemical assays, and histologic technique were used to assess the adverse effect of VGB in developing brain and its reversibility. RESULTS: At the end of 2 weeks' VGB administration: (a) there was a hyperactivity and a shortened latency to escape out of cool water; (b) white matter appeared hyperintense in T2 and diffusion-weighted MR images with 4 15% increases in T2; (c) microvacuolation, TUNEL-positive nuclei, and swollen axons were observed in the corpus callosum; (d) myelin staining indicated a reduction in myelination, as did the reduction in activities of myelin and oligodendrocyte-associated enzymes and the decrease in myelin basic protein on Western blots. Two weeks after stopping VGB administration: (a) MR images were normal, and microvacuolation was no longer in the white matter; (b) reduction in myelination reversed partially; (c) the T2 relaxation time remained elevated in the hypothalamus; and (d) the behavioral response remained abnormal. CONCLUSIONS: Long-term VGB administration to young rats causes brain injury, which recovers partially on its cessation. The observed cell death, disrupted myelination, and alterations in behavior indicate a need for further safety assessment in infants and children. PMID- 10840397 TI - Spatial extent of neuronal metabolic dysfunction measured by proton MR spectroscopic imaging in patients with localization-related epilepsy. AB - PURPOSE: To assess the spatial extent of the decrease in the neuronal marker N acetyl-aspartate (NAA) relative to creatine (Cr) in patients with localization related epilepsy, and to assess clinical differences between patients with and without widespread NAA/Cr reduction. METHODS: We studied 51 patients with localization-related epilepsy. Patients were divided into three groups according to the EEG investigation: (a) temporal lobe epilepsy (TLE, n = 21), (b) extratemporal lobe epilepsy (extra-TLE, n = 20), and (c) multilobar epilepsy (patients with a wider epileptogenic zone, n = 10). We acquired proton magnetic resonance (MR) spectrocopic imaging (1H-MRSI) of temporal and frontocentroparietal regions in separate examinations for both patients and controls. NAA/Cr values 2 standard deviations below the mean of normal controls were considered abnormal. RESULTS: Twenty-three (45%) patients including 12 with TLE had normal MR imaging including volumetric studies of the hippocampus. Forty nine (96%) patients had low NAA/Cr, indicating neuronal dysfunction in either temporal and/or extratemporal 1H-MRSIs; 38% of patients with TLE and 50% of patients with extra-TLE also had NAA/Cr reduction outside the clinical and EEG defined primary epileptogenic area. The NAA/Cr reduction was more often widespread in the multilobar group [six (60%) of 10] than in temporal or extratemporal groups [five (31%) of 16]. Nonparametric tests of (a) seizure duration, (b) seizure frequency, and (c) lifetime estimated seizures showed no statistically significant difference (p > 0.05) for TLE and extra-TLE patients with or without NAA/Cr reduction outside the seizure focus. CONCLUSIONS: Of patients with localization-related epilepsy, 40-50% have neuronal metabolic dysfunction that extends beyond the epileptogenic zone defined by clinical-EEG and/or the structural abnormality defined by MRI. PMID- 10840398 TI - Effects of gabapentin on brain GABA, homocarnosine, and pyrrolidinone in epilepsy patients. AB - PURPOSE: Gabapentin (GBP) was introduced as an antiepileptic drug (AED) and has been used in the management of neuropathic pain. We reported that daily dosing increased brain gamma-aminobutyric acid (GABA) in patients with epilepsy. This study was designed to determine how rapidly brain GABA and the GABA metabolites, homocarnosine and pyrrolidinone, increase in response to the first dose of GBP. METHODS: In vivo measurements of GABA, homocarnosine, and pyrrolidinone were made of a 14-cc volume in the occipital cortex by using a 1H spectroscopy with a 2.1 Tesla magnetic resonance spectrometer and an 8-cm surface coil. Six patients (four women) were studied serially after the first oral dose (1,200 mg) of GBP. Five patients (three women) taking a standard daily dose (range, 1,200-2,000 mg) of GBP were rechallenged with a single high dose (2,400 mg). RESULTS: The first dose of GBP increased median brain GABA by 1.3 mM (range, 0.4-1.8 mM) within 1 h. Homocarnosine and pyrrolidinone did not change significantly by 5 h. Daily GBP therapy increased GABA (0.5 mM; 95% CI, 0.2-0.9), homocarnosine (0.3 mM; 95% CI, 0.2-0.4), and pyrrolidinone (0.10 mM; 95% CI, 0.06-0.14). Rechallenging patients taking GBP daily increased median brain GABA by 0.4 mM (range, 0.3-0.5) within 1 h. CONCLUSIONS: GBP promptly elevates brain GABA and presumably offers partial protection against further seizures within hours of the first oral dose. Patients may expect to experience the anticonvulsant effects of increased homocarnosine and pyrrolidinone with daily therapy. PMID- 10840399 TI - Functional mapping of the insular cortex: clinical implication in temporal lobe epilepsy. AB - PURPOSE: We report the results of 75 intracortical electrical stimulations of the insular cortex performed in 14 patients during stereo-electroencephalography (SEEG) investigation of drug-resistant partial epilepsy. The insular cortex was investigated on electroclinical arguments suggesting the possibility of a perisylvian spread or a rapid multilobar diffusion of the discharges during video EEG. METHODS: In these 14 patients, 27 stereotactically implanted transopercular electrodes reached the insular cortex (11 the right insula, 16 the left insula). Square pulses of current were applied between the two deepest adjacent contacts of each transopercular electrode using low (1 Hz) or high-frequency (50 Hz) stimulation. Only symptoms evoked in the absence of afterdischarges were analyzed. RESULTS: Clinical responses were evoked in 10 of the 14 patients (in 20 of the 27 insular sites) and showed a clear topographic specificity inside the insular cortex. Viscerosensitive and visceromotor responses, similar to those evoked by temporomesial stimulation, were evoked by anterior insular stimulation and somesthetic sensation, similar to those evoked by opercular cortex stimulation, by posterior insular stimulation. CONCLUSIONS: The topographic organization of the induced responses within the insular cortex suggest that two different cortical networks, a visceral network extending to the temporomesial structures and a somesthetic network reaching the opercular cortex, are disturbed with stimulation of the anterior or the posterior insula, respectively. Thus ictal symptoms associated with the spread of the epileptic discharges to the insular cortex might be difficult to distinguish from those usually reported during temporomesial or opercular discharges. PMID- 10840401 TI - Cortical excitability in cryptogenic localization-related epilepsy: interictal transcranial magnetic stimulation studies. AB - PURPOSE: To assess whether single-and paired-pulse transcranial magnetic stimulation (TMS) can measure the interictal brain excitability of medicated patients with cryptogenic localization related epilepsy (CLE). Changes in the balance between excitation and inhibition are the core phenomena in focal epileptogenesis. TMS can assess this balance in the primary motor cortex. METHODS: We selected 18 patients with CLE and similar clinical features in whom we located the epileptogenic area reliably, with 11 age-and sex-matched healthy controls. For both motor cortices, we determined the threshold to TMS, the duration of the cortical silent period, and the corticocortical inhibition and facilitation curve. RESULTS: TMS was safe. The more antiepileptic drugs (AEDs) taken by the patients, the higher their threshold to TMS. The silent period duration failed to show significant changes. On paired TMS, a cluster analysis identified a homogeneous subgroup of patients (n = 7) who showed a significantly defective corticocortical inhibition and excess facilitation. With respect to the epileptogenic area, the phenomenon was bilateral in four of these patients, ipsilateral in two, and contralateral in one. The phenomenon was independent of AEDs and many other clinical variables. However, this patient group had a higher seizure frequency and a higher proportion of electroencephalograms (EEGs) showing interictal generalized epileptic discharges than the rest of the patients. CONCLUSION: Paired TMS provided a valuable pathophysiologic insight into the interictal excitatory state of the cortex in CLE. This method can potentially supply useful prognostic clinical information. PMID- 10840400 TI - Distinct behavioral and EEG topographic correlates of loss of consciousness in absences. AB - PURPOSE: To describe the behavioral and EEG topographic correlates of absences with 3-Hz generalized spike-waves and partitioned impairment of consciousness. METHODS: Two adult women had so-called "phantom" absences, characterized by brief and mild impairments of consciousness that were previously inconspicuous to both patient and physician. Neuropsychological examination was performed under video EEG monitoring during absence status. EEG topographic mapping of spike-wave discharges was obtained in the two cases. RESULTS: Only mild attentional and executive disturbances were observed during absence status despite prolonged discharges. Spike-wave bursts were associated with selective impairment in the initiation of response and self-generated action, whereas short-term storage of external information during discharges was fully preserved. This is consistent with a predominant involvement of frontomesial cortex demonstrated by topographic mapping of spike-wave discharges in the two cases. By contrast, in two other patients with typical absences and a complete lack of retention for information given during the discharges, topographic mapping found a more lateral frontal involvement by spike-wave activity. CONCLUSIONS: Different types of absence seizures may impair distinct components of conscious behavior. A predominant involvement of frontomesial thalamocortical circuitry may underlie an "inconspicuous" disorder of consciousness as seen in phantom absences with selective loss of initiation and goal-oriented behavior, whereas involvement of more lateral frontal areas in typical absences may additionally disrupt working memory processes. PMID- 10840404 TI - Early recognition of benign partial epilepsy in infancy. AB - PURPOSE: The aim of this study is to determine how precisely we can recognize the outcome in infants with epilepsy beginning in the first year of life. METHODS: We performed a prospective 5-year follow-up study on 63 patients who developed epilepsy in the first year of life. We first judged that patients met the criteria of "possible benign partial epilepsy in infancy (BPEI)" on enrollment in this study. At 2 years of age, we reevaluated the seizure and developmental outcome in the patients who were diagnosed as having "possible BPEI." We finally judged that patients met the criteria of "definite BPEI" at age 5 years. "Possible BPEI" was defined as epilepsy meeting all the following conditions: (a) complex partial seizures and/or secondarily generalized seizures; (b) normal psychomotor development and neurologic findings before onset; (c) normal interictal electroencephalograms; (d) normal cranial computed tomography (CT) and magnetic resonance imaging (MRI) findings; and (e) no seizures during the first 4 weeks of life. "Definite BPEI" was defined as epilepsy meeting all the following criteria in addition to those of "possible BPEI": (a) normal psychomotor development beyond age 5 years, and (b) no seizures beyond age 2 years. RESULTS: Thirty-two of the 63 patients met the inclusion criteria completely and were included in the "possible BPEI" group. Twenty-five of the 32 patients completed the 5-year follow-up. At age 2 years, four patients were excluded from the "possible BPEI" group because of seizure recurrence and/or delayed development. By age 5 years, one had a recurrence of seizures, and another exhibited mildly delayed psychomotor development. We finally diagnosed 19 patients as having "definite BPEI." "Definite BPEI" accounted for 76% of the patients diagnosed as having "possible BPEI" at the first presentation and 90% of those who met the conditions on reevaluation at age 2 years. CONCLUSIONS: Recognition of BPEI is possible, to some extent, at the first presentation, and reevaluation at age 2 years is useful for a more precise diagnosis. PMID- 10840402 TI - Markedly elevated nitrate/nitrite levels in the cerebrospinal fluid of children with progressive encephalopathy with edema, hypsarrhythmia, and optic atrophy (PEHO syndrome). AB - PURPOSE: To compare the levels of brain nitric oxide production in patients with PEHO or PEHO-like syndrome and in controls with other neurologic disease. METHODS: Nitric oxide metabolites, nitrates, and nitrites (NNx), were measured in the cerebrospinal fluid (CSF) of children with PEHO syndrome or PEHO-like syndrome, and in controls with other neurologic diseases. RESULTS: The NNx levels were markedly higher in both PEHO (mean, 48 microM; p < 0.001) and PEHO-like (22 microM; p < 0.003) patients as compared with the controls (6 microM), but did not correlate with age or with brain atrophy or CSF levels of insulin-like growth factor-1 (IGF-1). CONCLUSIONS: Our findings suggest that in PEHO syndrome, production of nitric oxide is markedly increased, suggesting that nitric oxide is involved in the pathologic phenomena (i.e., seizures and neurodegeneration) of the disease. PMID- 10840403 TI - Lamotrigine in pregnancy: pharmacokinetics during delivery, in the neonate, and during lactation. AB - PURPOSE: To investigate the pharmacokinetics of lamotrigine (LTG) during delivery, during the neonatal period, and lactation. METHODS: High-performance liquid chromatography was used to determine plasma and milk levels of LTG in nine pregnant women with epilepsy treated with LTG, and plasma levels in their 10 infants. Samples were obtained at delivery, the first 3 days postpartum, and at breast-feeding 2-3 weeks after delivery. RESULTS: At delivery, maternal plasma LTG concentrations were similar to those from the umbilical cord, indicating extensive placental transfer of LTG. There was a slow decline in the LTG plasma concentration in the newborn. At 72 h postpartum, median LTG plasma levels in the infants were 75% of the cord plasma levels (range, 50-100%). The median milk/maternal plasma concentration ratio was 0.61 (range, 0.47-0.77) 2-3 weeks after delivery, and the nursed infants maintained LTG plasma concentrations of approximately 30% (median, range 23-50%) of the mother's plasma levels. Maternal plasma LTG concentrations increased significantly during the first 2 weeks after parturition, the median increase in plasma concentration/dose ratio being 170%. CONCLUSIONS: Our data demonstrate a marked change in maternal LTG kinetics after delivery, possibly reflecting a normalization of an induced metabolism of LTG during pregnancy. LTG is excreted in considerable amounts in breast milk (the dose to the infant can be estimated to >/=0.2-1 mg/kg/day 2-3 weeks postpartum), which in combination with a slow elimination in the infants, may result in LTG plasma concentrations comparable to what is reported during active LTG therapy. No adverse effects were observed in the infants, however. PMID- 10840407 TI - The coordinated psychosocial and neurologic care of children with seizures and their families. AB - SUMMARY: As the medical and surgical management of epilepsy continues to advance, issues associated with the quality of life of patients and their families can be addressed. Whenever associated with other handicaps, such as learning disabilities, attentional or behavioral disorders, and problems in psychological adjustment, dual-diagnosis issues must be identified. To provide comprehensive care for children with epilepsy, a team approach to psychosocial assessment and treatment must be provided and coordinated with neurologic care. When the age related needs in the life stage of the individual and family are identified, the best possible adaptation of the patient and his or her family can be supported. PMID- 10840405 TI - Focal cortical-subcortical calcifications (FCSCs) and epilepsy in the Indian subcontinent. AB - PURPOSE: A focal cortical-subcortical calcification (FCSC) is a common finding on computed tomography (CT) in individuals with focal or generalized seizures in the Indian subcontinent. We sought to determine the relation of FCSCs to epilepsy by comparing the lobe of seizure origin by electroclinical and CT evaluations and to study the nature and severity of epilepsy associated with FCSCs. METHODS: The relation of these FCSCs to epilepsy/seizures was studied in 40 patients, seen for the first time to the neurology outpatient department of a tertiary care hospital. An attempt was made to classify seizures and determine their lobe of origin based on clinical-electroencephalographic (EEG) criteria of the International League Against Epilepsy (ILAE). The clinical lobe of origin was compared with the location of the FCSC on CT scan. In addition, records of the CT unit of the same hospital were reviewed retrospecitvely, to identify cases with an FCSC and their referral diagnoses. RESULTS: Thirty-one (77.5%) patients with FCSCs were considered to have localization-related epilepsy (frontal lobe epilepsy, 20; temporal lobe epilepsy, three; parietal lobe epilepsy, one; occipital lobe epilepsy, three; and definitely localization related but having ambiguous localization features, four) based on ictal semiology and EEG studies. Other ILAE categories in the cohort included epilepsy without unequivocal focal or generalized features (four patients; 10%), isolated seizures (one patient; 2.5%), juvenile absence epilepsy (one patient; 2.5%), and insufficient data to classify epilepsy (three patients; 7.5%). Radiologic sites for FCSCs included frontal (20; 50%), temporal (six; 15%), parietal (seven; 17.5%), and occipital (seven; 17.5%). Electroclinical and radiologic data were congruent in localizing and lateralizing seizures in 22 (55%) patients. The FCSC was truly incidental in one patient with juvenile absence epilepsy. Discordance between the clinical and radiologic localizations was noted in five (12.5%) instances. Magnetic resonance imaging (MRI) did not reveal additional lesions corresponding to lobes of origin as determined by electroclinical analysis. Discordance was surmised to be a result of seizure spread from a silent region to symptomatic cortex. In 12 (30%) patients, electroclinical and radiologic congruence could not be ascertained because ictal descriptions were either inadequate or ambiguous, and EEG findings were noncontributory. Review of 4,452 CT scans of brain performed in the CT unit revealed 29 (0.65%) cases with FCSCs in individuals with nonseizure disorders, that could be labeled as incidental. CONCLUSIONS: An FCSC is an important radiologic finding in localization-related epilepsy in the Indian subcontinent. The severity of epilepsy ranges from asymptomatic cases to daily seizures. PMID- 10840406 TI - An assessment of epilepsy patients' knowledge of their disorder. AB - PURPOSE: Patient education is an effective component of comprehensive care. Studies assessing patient's knowledge of their epilepsy are scarce. We report the first objective study evaluating knowledge of epilepsy patients referred to an American tertiary care center. METHODS: Two hundred twenty epilepsy patients referred to an epilepsy center completed a knowledge questionnaire. The questionnaire included topics related to safety, compliance, and legal issues of driving and employment. Questionnaire scores were correlated with demographics, number of years with epilepsy, and educational background. RESULTS: Of 220 patients, 175 were included in study analysis. Thirteen percent (n = 28) were excluded because of the diagnosis of nonepileptic seizures, and 8% (n = 17) were excluded because of having a diagnosis other than epilepsy. The average age and number of years with epilepsy was 34.7 +/- 13 and 14. 4 +/- 13.1, respectively. Neither age (r = 0.20, p /=50% of their patients, but a minority of physicians (n = 122) refer <50% of their patients. Differences between the two groups existed in three of the four research questions asked: who initiates AED therapy, comfort level, and percentage of patients referred to a neurologist. Influence of managed care on decision making was not different between the two groups. CONCLUSIONS: A minority of primary care physicians rate themselves very comfortable with seizure patients. These same physicians refer a minority of their patients to a neurologist. As a whole, however, primary care physicians refer a majority of their seizure patients to a neurologist. Neurologists evaluate most seizure patients because most primary care physicians claim not to be extremely comfortable with evaluation and treatment of seizures. We conclude that neurologists play an essential role in the treatment of most seizure patients. PMID- 10840409 TI - Health-related quality of life outcome in medically refractory epilepsy treated with anterior temporal lobectomy. AB - PURPOSE: A prospective study to investigate health-related quality of life (HRQOL) outcome in patients with temporal lobe epilepsy treated with anterior temporal lobectomy (ATL). METHODS: The majority of the patients with medically refractory focal epilepsy had Quality of Life in Epilepsy-89 (QOLIE-89) assessment at the time of prolonged video/EEG monitoring as part of their presurgical evaluation. Thirty-seven patients who were not treated surgically constituted the control group, and 53 patients who underwent ATL made up the surgery group. Both control and surgery groups had HRQOL assessment repeated at approximately 1-and 2-year intervals. Repeated measures analysis of variance (ANOVA) was used to test for differences between the two groups. RESULTS: For the overall score and almost every scale, the surgery group had a higher baseline mean than the control group. Because of this baseline difference, change scores were used in further analysis. The overall score and 10 of 17 scales in QOLIE-89 showed significant HRQOL improvement after ATL, and the improvement was significant relative to score changes of the nonsurgical comparison group. Scores improved in overall QOL, emotional well-being, attention/concentration, language, social isolation, health perception, role limitations-physical, work/drive/social, health discouragement, and seizure worry. For the first five scales, there was group-time interaction; the improvement was significantly more on the 2-year than on the 1-year follow-up. When the surgery patients were divided into four categories (class IA-, completely seizure free; class IA+, seizure free with aura; class II, rare seizures; class III, worthwhile improvement in seizure control; and class IV, no improvement), the improved HRQOL in the surgery group was almost entirely contributed by the class IA- outcome patients who were totally seizure free. The class IA+ patients with continuing aurae and class II/III/IV patients had no significant improvement in their overall HRQOL scores at 1-or 2-year follow-up. CONCLUSIONS: Overall score and 10 of the 17 scales of QOLIE-89 significantly improved in patients with medically refractory temporal lobe epilepsy after ATL. For some scales, there was delay in the improvement to manifest. The HRQOL improvement was related to achieving an entirely seizure-free status (i.e., no seizures or aurae postoperatively). PMID- 10840410 TI - Relationships between seizure severity and health-related quality of life in refractory localization-related epilepsy. AB - PURPOSE: To evaluate relationships between self-report measures of seizure severity and health-related quality of life (HRQOL) in people with refractory localization-related epilepsy. METHODS: A sample of 340 adults enrolled in a seven-center, prospective study of resective epilepsy surgery completed baseline questionnaires that included the Quality of Life in Epilepsy (QOLIE)-89 and a seven-item adaptation of the National Hospital Seizure Severity Scale. Associations between QOLIE-89 summary measures and both the total seizure severity scale score and individual seizure severity items were assessed, after adjustment for seizure frequency. RESULTS: The seizure severity measure had adequate scale score variability and reliability in this sample. Correlations between individual items in the scale did not exceed 0. 43. Product-moment partial correlations between the seizure severity scale and QOLIE-89 summary measures ranged from -0.17 to -0.29 (all p values <0.01). Of the seven seizure severity items, the average time before individuals perceived they were "really back to normal" after their seizures was broadly related to all domains of HRQOL (r values ranged from -0.16 to -0.30; p values <0.01). Severity of injury during seizures was the only other item having more than minimal associations with HRQOL, and it was selectively related to the physical health measure. Higher frequency of falls during seizures was modestly related to less employment. CONCLUSIONS: This seizure severity measure assesses constructs that are generally distinct from HRQOL, except for moderate and broad associations between HRQOL and patient's perceptions of the average duration of recovery time after seizures. Recovery time may potentially be a useful clinical indicator of seizure severity that reflects meaningful impairment of HRQOL in adults with frequent seizures. PMID- 10840411 TI - Validation of a new quality of life measure for children with epilepsy. AB - PURPOSE: There is no adequate measure of health-related quality of life (HRQOL) specifically for children with epilepsy. The aim of this study was to develop an epilepsy-specific HRQOL questionnaire for children, covering five domains: physical function, emotional well-being, cognitive function, social function, and behavior. Second, we aimed to demonstrate the instrument's reliability and validity, and its sensitivity to differences in epilepsy severity. METHODS: The subjects were guardians of children with refractory epilepsy, whose syndrome had been defined by using video-EEG monitoring. Each family completed the developed epilepsy-specific HRQOL scale for children and two standard, generic measures of HRQOL. RESULTS: The results indicated that each of the scales of the questionnaire had good internal consistency reliability. Furthermore, each scale correlated more highly with theoretically similar scales on established, generic health measures than with theoretically dissimilar scales (construct validity). The sensitivity of the questionnaire to differences in epilepsy severity also was demonstrated. As seizure severity increased, HRQOL subscale scores decreased, independent of age, gender, age of seizure onset, and IQ. Further, there was a negative relation between the number of antiepileptic medications taken and measures of memory and language performance, which was independent of age, gender, age of seizure onset, IQ, and seizure severity. CONCLUSIONS: This study demonstrated that the developed HRQOL instrument is a reliable and valid measure and is sensitive to differences in epilepsy. These results indicate that this new instrument may be a viable medical or surgical outcome measure for children with epilepsy. PMID- 10840412 TI - Complete androgen blockade for prostate cancer: what went wrong? AB - PURPOSE: We summarized and critically assessed all available data from phase III clinical trials on complete androgen blockade versus surgical or medical castration alone. MATERIALS AND METHODS: Published results in journals and abstracts of phase III trials, and published meta-analyses were reviewed. We also reviewed quality of life and toxicity issues associated with the addition of antiandrogens to medical or surgical castration. Finally, we discuss the original rationale for complete androgen blockade in the context of current knowledge. RESULTS: A total of 27 clinical trials using various combinations of androgen deprivation were identified, of which 3 showed a statistically significant benefit for the complete androgen blockade arm. There were 5 publications of meta analyses that each used different selection criteria for the inclusion of studies in the final analysis. Toxicity and quality of life have not been widely investigated in prospective fashion but the available data suggest a higher toxicity rate and decreased quality of life with complete androgen blockade. CONCLUSIONS: The extensive body of data does not support routine use of antiandrogens in combination with medical or surgical castration as first line hormonal therapy in patients with metastatic prostate cancer. PMID- 10840413 TI - Nocturnal enuresis in men with an orthotopic ileal reservoir: urodynamic evaluation. AB - PURPOSE: We evaluated the urodynamic features of enuretic and continent patients with an orthotopic neobladder. MATERIALS AND METHODS: Included in our study were 100 men with an orthotopic hemi-Kock or W neobladder and a minimum followup of 1 year. Of the patients 50 were completely continent day and night, and 50 had enuresis without evidence of an underlying organic etiology, such as stones, reflux or urethral stricture. RESULTS: Univariate analysis showed significantly higher pressure and a larger volume of post-void residual urine in the men with enuresis. In addition, maximum urethral pressure, maximum flow and compliance were decreased in the enuretic group compared to the continent group. Multivariate analysis revealed that post-void residual urine volume, frequency and maximum amplitude of uninhibited contractions are the most critical parameters affecting nocturnal continence. CONCLUSIONS: The cystometric parameters significantly associated with nocturnal enuresis in patients with an orthotopic reservoir are post-void residual urine volume, frequency and maximum amplitude of uninhibited contractions. Urethral pressure and flow parameters did not sustain significance on multivariate analysis. PMID- 10840414 TI - Laparoscopic partial adrenalectomy in patients with hereditary forms of pheochromocytoma. AB - PURPOSE: Patients with von Hippel-Lindau disease are predisposed to multiple bilateral adrenal pheochromocytoma. In these patients partial adrenalectomy may preserve adrenocortical function and avoid the morbidity associated with medical adrenal replacement. We report our experience with such cases. MATERIALS AND METHODS: Laparoscopic partial adrenalectomy was performed in patients with von Hippel-Lindau disease and pheochromocytoma when there was evidence of normal adrenocortical tissue on preoperative imaging or intraoperative examination. Suture ligature or a harmonic scalpel was used to excise the tumors, leaving a 2 to 3 mm. margin of normal tissue. RESULTS: Two patients underwent laparoscopic partial adrenalectomy and 1 laparoscopic bilateral partial adrenalectomy with preservation of normal adrenocortical tissue. Seven pheochromocytomas were removed. Laparoscopic ultrasound was essential for localizing 2 pheochromocytomas that were not visualized by the camera. Median operative time was 324 minutes, blood loss 100 cc and parenteral narcotic requirement 22 mg. morphine equivalents. No patient required hydrocortisone replacement. There has been no pheochromocytoma recurrence during short-term followup. CONCLUSIONS: Laparoscopic partial adrenalectomy is technically feasible in patients with a hereditary form of pheochromocytoma, and may preserve adrenocortical function. Laparoscopic ultrasound was necessary to identify 2 of the 7 pheochromocytomas removed. PMID- 10840415 TI - Caliceal diverticular calculi: is there a role for metabolic evaluation? AB - PURPOSE: We report our experience with the treatment and incidence of metabolic abnormalities in patients presenting with caliceal diverticular stones. MATERIALS AND METHODS: We retrospectively evaluated 49 patients with caliceal diverticular stones (group 1) and 44 with simple renal stones (group 2). Each group successfully underwent percutaneous treatment. Mean stone size was 1.7 and 2.5 cm. in groups 1 and 2, respectively. Metabolic evaluation was available in 25 group 1 and 22 group 2 patients. Mean followup was 73.2 and 70. 8 months, respectively. RESULTS: We achieved a stone-free rate of 95. 9% in group 1 and 100% in group 2. There was no metabolic abnormality in 75% of the group 1 patients, while 12% had type II absorptive hypercalciuria, 8% hyperuricosuric hypercalciuria and 4% hyperoxaluria. There were no metabolic abnormalities in 22.7% of the group 2 patients, while 9%, 18% and 9% had types I to III absorptive hypercalciuria, respectively, 13.6% hyperuricosuric hypercalciuria, 13.6% hyperoxaluria, 4.5% hypocitruria and 9% type II absorptive hypercalciuria associated with hypocitruria. CONCLUSIONS: Our results reveal a low incidence of associated metabolic abnormalities in patients with caliceal diverticular stones. Thus, we believe that metabolic abnormalities do not promote caliceal diverticular calculous formation. PMID- 10840416 TI - Geographic variation and environmental risk factors for the incidence of initial kidney stones in patients with spinal cord injury. AB - PURPOSE: Although geographic variability in cases of kidney stones, primarily calcium stones, is reported in the general population, little is known about geographic variability in subjects with spinal cord injury, in whom struvite stones predominate. We examined regional variation in the incidence rate of initial kidney stones and clarified the contributing factors in a spinal cord injury cohort. MATERIALS AND METHODS: We used data from the national spinal cord injury database between 1986 and 1999 on 7,784 participants from 21 spinal cord injury centers with 24,492 person-years of followup accumulated and 286 incident stone cases. A multilevel Poisson model was constructed to evaluate the ecological effects of latitude, air temperature, water hardness and sunlight index on stone formation while controlling for individual factors, including participant age, race, gender, severity of injury and bladder management. RESULTS: The incidence rate was significantly greater in the southeast and tended to increase with decreasing latitude, similar to the geographic association with kidney stones in the general population. This finding was not explained by differences in individual risk factors. Decreasing water hardness had the strongest effect on stones during year 1 after injury (relative risk 0.6, p <0. 001), whereas average annual temperature had the strongest association with stones after year 1 (relative risk 1.1 per 1C increase, p = 0.03). The sunlight index had no association. CONCLUSIONS: Our study implies that the increased stone risk in spinal cord injured subjects is potentially preventable by modifying environmental exposure. Etiological factors may be similar to those for kidney stones in the general population. PMID- 10840417 TI - Selective nonoperative management of blunt grade 5 renal injury. AB - PURPOSE: We determined the feasibility of a nonoperative approach to blunt grade 5 renal injury. MATERIALS AND METHODS: We retrospectively reviewed the records of all patients with grade 5 renal injury who presented to our level 1 trauma center from 1993 to 1998. Those treated nonoperatively and surgically were assigned to groups 1 and 2, respectively. Each group was compared with respect to the initial emergency department evaluation, computerized tomography findings, associated injuries, duration of hospital stay and intensive care unit stay, transfusion requirements, complications and followup imaging. RESULTS: Of 218 renal injuries evaluated 13 were grade 5. In group 1, 6 patients were treated nonoperatively and in group 2, 7 underwent exploration. Each group had similar average hospitalization (12.0 and 12.8 days, respectively). Patients in group 1 had fewer intensive care unit days (4.3 versus 9.0), significantly lower transfusion requirements (2.7 versus 25.2 units, p = 0.0124) and fewer complications during the hospital course. Followup computerized tomography of nonoperatively managed cases revealed functioning renal parenchyma with resolution of retroperitoneal hematoma. CONCLUSIONS: Conservative management of blunt grade 5 renal injury is feasible in patients who are hemodynamically stable at presentation. PMID- 10840418 TI - Comparison of retroperitoneoscopic nephrectomy with open surgery for tuberculous nonfunctioning kidneys. AB - PURPOSE: We describe, define and evaluate the role of retroperitoneoscopic nephrectomy for tuberculous nonfunctioning kidneys, and compare the results with those of open nephrectomy in similar cases in a nonrandomized study. MATERIALS AND METHODS: Beginning in July 1994, 9 patients underwent retroperitoneoscopic nephrectomy for tuberculous nonfunctioning kidneys at our center. Data obtained from the records of these patients were compared with those of 9 who underwent open nephrectomy for a similar indication during the same period. Retroperitoneoscopic nephrectomy was initially performed by kidney dissection followed by ligation of the hilar vessels. The technique was subsequently modified and the vessels controlled before dissecting the kidney. Various parameters were compared and statistical analysis was done. RESULTS: The 2 groups were similar in regard to patient age, gender and side of disease. Retroperitoneoscopic nephrectomy was successful in 7 of the 9 patients. Although 2 of our initial patients required conversion to open surgery, the remaining 7 successfully underwent retroperitoneoscopic nephrectomy after modifying the technique. Mean operative time was slightly greater in the retroperitoneoscopy than in the open surgery group (103.3 versus 92.2 minutes). Mean blood loss was less in the retroperitoneoscopy group (101.4 versus 123.3 ml.), mean hospital stay plus or minus standard deviation was significantly shorter (3.2 +/- 0.83 versus 8.88 +/- 3.37 days) and mean time to return to work was significantly less (3 versus 7 weeks). Mean analgesic requirement for opioids and diclofenac sodium was also lower in the retroperitoneoscopic nephrectomy group (0 versus 1.44 +/- 0.72 and 3.8 +/- 1.3 versus 4.3 +/- 1.2 doses, respectively). Minor complications developed in only 2 retroperitoneoscopy cases. CONCLUSIONS: Tuberculosis has been considered a contraindication to retroperitoneoscopic nephrectomy due to a high conversion rate. However, we believe that our modified technique of retroperitoneoscopic nephrectomy is a viable option for managing tuberculous nonfunctioning kidneys. The conversion rate is lower than previously reported. Comparing our results with those of open nephrectomy shows that retroperitoneoscopic nephrectomy is beneficial in all respects except for slightly longer operative time. Because of the benefits of minimally invasive surgery, this approach should be considered in such cases. PMID- 10840419 TI - Influence of tumor stage, size, grade, vascular involvement, histological cell type and histological pattern on multifocality of renal cell carcinoma. AB - PURPOSE: We assessed the influence of clinical and pathological factors on multifocality of renal cell carcinoma. MATERIALS AND METHODS: Between June 1995 and September 1999 radical nephrectomy was performed in 71 men and 32 women with a mean age of 56.5 years. The 103 removed kidneys with renal cell carcinoma were sectioned at 3 mm. intervals and inspected microscopically for satellite carcinomas. We evaluated pathological stage, grade, cell type, histological pattern, vascular involvement, tumor size and the incidence of multifocality. To determine cell type we used several classification systems. RESULTS: The primary tumor was 2 to 20 cm. (mean plus or minus standard deviation 7.10 +/- 3.48). Overall satellite carcinomas were present in 22 of the 103 cases (21.4%). When the predominant lesion was 5 cm. or smaller, the incidence of multifocality was 19%. The incidence of multifocality was statistically higher in patients with stage pT3 than in those with stage pT1 or pT2 disease (p = 0.022). Multiple logistic regression analysis demonstrated that only primary tumor pathological stage was a significant predictor of renal cell carcinoma multifocality in stages T3 versus T1 and T3 versus T2 cancer (odds ratio 3.45, 95% confidence interval 1.15 to 10.39 and 5.75, 1.31 to 25.29, respectively). Other parameters, such as tumor size, grade, vascular invasion, cell type and histological pattern, did not correlate with multifocality. CONCLUSIONS: Our results imply that primary tumor stage is a significant factor for multifocal disease. Therefore, more precise preoperative staging of the primary lesion is required if nephron sparing surgery is indicated. PMID- 10840420 TI - Clinical efficacy of combined lithoclast and lithovac stone removal during ureteroscopy. AB - PURPOSE: Pneumatic lithotripsy has proved to be an extremely safe, efficient and low cost intracorporeal fragmentation modality. Unfortunately proximal migration of fragments into inaccessible areas in the intrarenal collecting system is a potential limitation during ureteroscopic procedures. Moreover, the lack of an efficient mechanism of stone retrieval further limits the widespread application of this technique. The newly developed Lithovacdagger suction device was designed to be combined with pneumatic lithotripsy during ureteroscopic stone removal. We evaluated the effectiveness of this combination for overcoming these limitations. MATERIALS AND METHODS: Between February and December 1998, 21 patients underwent pneumatic lithotripsy of ureteral calculi combined with use of the Lithovac suction probe. Stone area was 20 to 320 mm.2 (mean 84). Of the stones 71% and 29% were in the distal, and proximal and/or mid ureter, respectively. We used a 0.8 mm. pneumatic lithotripsy probe placed through a 4.8Fr Lithovac probe at a pulse frequency of 12 Hz. and pressure of 2 atmospheres. RESULTS: Mean operative time was 42 minutes and the stone fragmentation rate was 100%. There were no complications in our series and no proximal migration of fragments. The overall stone-free rate was 95% at 3-month followup. CONCLUSIONS: Using the Lithovac suction device greatly facilitates pneumatic lithotripsy during ureteroscopic stone removal. This combination not only prevents fragment migration, but also aids in maintaining a clear endoscopic field of view, allowing efficient, safe and effective stone fragmentation. PMID- 10840421 TI - Computerized tomography cystography for the diagnosis of traumatic bladder rupture. AB - PURPOSE: We present our experience with computerized tomography (CT) cystography for diagnosing bladder rupture in patients with blunt abdominal and pelvic trauma, and compare the results of CT cystography with those of surgical exploration. MATERIALS AND METHODS: We identified all patients with blunt trauma diagnosed with bladder rupture from 1992 to September 1998. We reviewed the radiology computerized information system for all CT cystography performed to evaluate blunt trauma during the same period. We also reviewed the medical records and pertinent radiographic studies of patients with bladder rupture who underwent CT cystography as part of the hospital admission evaluation. Operative and radiographic findings were compared. RESULTS: CT cystography was performed in 316 patients as part of the initial evaluation of blunt trauma. Of the 44 patients with the ultimate diagnosis of bladder rupture CT cystography revealed bladder rupture in 42, while 23 of the 28 (82%) who underwent formal bladder exploration had operative findings that exactly matched the CT cystography interpretation in terms of the presence and type of rupture. In the 316 patients CT cystography detected bladder rupture with an overall sensitivity and specificity of 95% and 100%, respectively. For intraperitoneal rupture sensitivity was 78% and specificity was 99%. CONCLUSIONS: CT cystography provides expedient evaluation of bladder rupture due to blunt trauma and has accuracy comparable to that reported for plain film cystography. We recommend CT cystography over plain film cystography in patients undergoing CT for other injuries associated with blunt trauma. PMID- 10840422 TI - Weekly mitomycin C followed by monthly bacillus Calmette-Guerin or alternating monthly interferon-alpha2B and bacillus Calmette-Guerin for prophylaxis of recurrent papillary superficial bladder carcinoma. AB - PURPOSE: We evaluated alternatives to bacillus Calmette-Guerin (BCG) monotherapy using a new combination of chemotherapy and immunotherapy for recurrent superficial bladder carcinoma. MATERIALS AND METHODS: A total of 236 patients with frequently recurrent stage Ta or T1 bladder tumors were enrolled in our prospective, randomized, multicenter Finnbladder IV study. The initial mitomycin C instillation was instilled in all patients perioperatively after transurethral resection, followed by 4 weekly instillations of mitomycin C. Thereafter patients were randomized to receive monthly for up to 1 year BCG only or interferon alpha2b and BCG alternating monthly. Primary end points were time to initial recurrence, recurrence rate (number of recurrences per patient-year) and recurrence index (number of recurrent tumors per patient-year). RESULTS: Of the 236 randomized patients 205 were eligible for study with a median overall followup of 30.7 months. Monthly BCG was superior to alternating monthly interferon-alpha and/or BCG with respect to time to initial recurrence (log rank test p <0.00001) as well as recurrence rate (0.4 versus 0.9, p <0.00001) and index (0.9 versus 3.0, p <0.00001). Side effects were limited. CONCLUSIONS: Monthly BCG given for up to 1 year preceded by perioperative and an additional 4 weekly mitomycin C instillations is a well tolerated mode of instillation therapy, providing excellent tumor control comparable to that of the best reported instillation regimens. No benefit was obtained by alternating interferon alpha2b with BCG. PMID- 10840423 TI - Gemcitabine plus cisplatin for advanced transitional cell carcinoma of the urinary tract: a phase II multicenter trial. AB - PURPOSE: We determined the activity and toxicity of gemcitabine plus cisplatin in patients with inoperable or metastatic transitional cell carcinoma of the urinary tract. MATERIALS AND METHODS: A total of 54 patients with transitional cell carcinoma, measurable disease and Eastern Cooperative Oncology Group performance status 2 or greater were enrolled in this multicenter phase II trial. Previous adjuvant or neoadjuvant therapy for locally advanced disease was acceptable if it had been completed more than 1 year before study entry. Every 4 weeks patients received 1,000 mg./m.2 gemcitabine intravenously on days 1, 8 and 15, and 70 mg./m.2 cisplatin intravenously on day 2. RESULTS: All patients were evaluable for response and toxicity. Notably only 7 of the 54 patients (13%) previously received chemotherapy in an adjuvant or neoadjuvant setting. Overall we observed 26 objective responses (48%), of which 15% were complete. Median time to progression was 23 weeks and median survival was 54 weeks. Treatment was well tolerated. The main toxicities were leukopenia (grade 3 in 28% and grade 4 in 11% of patients), anemia (grade 3 in 34% and grade 4 in 6%) and thrombocytopenia (grade 3 in 14% and grade 4 in 6%). Other relevant side effects were nausea and vomiting in 20% of cases, fever in 24%, alopecia in 22%, renal failure in 7.4% and mucositis in 2%. CONCLUSIONS: Combined cisplatin plus gemcitabine is highly active in advanced transitional cell carcinoma of the urinary tract with manageable toxicity. The response rate, time to treatment failure and overall survival appeared to be comparable to those achieved with combined methotrexate, vinblastine, doxorubicin and cisplatin. Conversely toxicity appeared lower. Evaluation of this regimen in randomized studies with methotrexate, vinblastine, doxorubicin and cisplatin is strongly suggested. PMID- 10840424 TI - Scrotal dog bites. AB - PURPOSE: Dog bites to the scrotum are rare but they potentially result in morbidity if improperly managed. MATERIALS AND METHODS: Between 1991 and 1999 we treated 4 men and 3 boys with dog bites to the scrotum. All 7 patients presented to the emergency department shortly after the injury. Of the 4 adults 3 were ingesting alcohol and 2 were obviously intoxicated, and 1 had a T4 spinal cord injury and was bitten during sleep. Of the children 2 were apparently bitten without provocation, while a 5-year-old boy was bitten when the family dog was disturbed while eating. RESULTS: All wounds were explored, irrigated and debrided. There was no involvement of the testes or spermatic cord. Each wound was closed primarily and 5 healed without sequelae. The spinal cord injured man had partial dehiscence of the incision and in another man superficial hematoma required drainage. Each patient received antibiotics and tetanus prophylaxis but none required rabies inoculation. CONCLUSIONS: Although there are reports of devastating scrotal injuries from dog bites, most such wounds may be treated by careful inspection for intrascrotal injuries followed by debridement and closure. Antimicrobial prophylaxis should be administered, as for any bite wound. PMID- 10840425 TI - A prospective randomized trial comparing transurethral resection of the prostate and laser therapy in men with chronic urinary retention: The CLasP study. AB - PURPOSE: We assessed the effectiveness of laser therapy versus transurethral prostatic resection in men with symptomatic chronic urinary retention secondary to benign prostatic enlargement. MATERIALS AND METHODS: This trial was multicenter, pragmatic and randomized. Analysis was done by intent to treat. Laser therapy involved neodymium:YAG noncontact visual prostate ablation, while transurethral prostatic resection was performed by standard electroresection. Patients were included in our study if they reported moderate to severe lower urinary tract symptoms with an International Prostate Symptom Score (I-PSS) of 8 or more, benign prostatic enlargement and a persistent post-void residual urine volume of more than 300 ml. Followup was 7.5 months. Primary outcome measures included the I-PSS, I-PSS quality of life score, maximum urinary flow and post void residual urine volume. Secondary outcome measures included treatment failure, complications, hospital stay and catheterization time. RESULTS: A total of 82 patients agreed to be randomized to receive laser therapy (38) or transurethral prostatic resection (44). There were significant improvements in all primary outcomes in each group from randomization to followup. Transurethral prostatic resection was significantly better than laser therapy for I-PSS and maximum urinary flow values (p = 0.035 and 0.029, respectively) but there were no differences in post-void residual urine volume and I-PSS quality of life score between the groups. We noted significantly more treatment failures with laser therapy than resection (8 versus 0, p = 0.0014), although only 3 patients required resection after laser therapy because of persistent symptoms. In addition, hospital stay after resection was 2-fold that after laser therapy (ratio of geometric means 2.01, 95% confidence interval 1.54 to 2.61, p <0.0001). However, time to catheter removal was 9 times longer in the laser therapy group (p <0. 0001). Complication rates were significantly higher for transurethral prostatic resection (chi-square 5.05, 1 df, p = 0.025). CONCLUSIONS: Transurethral prostatic resection is more effective than laser ablation in men with chronic urinary retention in terms of symptom score, maximum urinary flow and failure. However, men who underwent resection had significantly more treatment complications and were hospitalized longer than those who received laser therapy. This finding implies that laser ablation therapy may have a role in patients at higher risk who are willing to accept a lower level of effectiveness in exchange for decreased complication rates and hospital stay. PMID- 10840427 TI - Comparison of therapies for lower urinary tract symptoms. PMID- 10840426 TI - A randomized trial comparing transurethral resection of the prostate, laser therapy and conservative treatment of men with symptoms associated with benign prostatic enlargement: The CLasP study. AB - PURPOSE: We evaluated the effectiveness of a new technology (noncontact laser therapy) versus that of standard surgery (transurethral prostatic resection) and conservative management for lower urinary tract symptoms associated with benign prostatic enlargement. MATERIALS AND METHODS: Men with uncomplicated lower urinary tract symptoms, that is no acute or chronic urinary retention, were randomized to receive laser therapy with a noncontact, side firing neodymium:YAG probe, standard transurethral prostatic resection or conservative management, including monitoring without active intervention, in a large multicenter pragmatic randomized controlled trial called the CLasP study. Primary outcomes were International Prostate Symptom Score (I-PSS), maximum urinary flow rate, a composite measure of success based on I-PSS and maximum urinary flow rate categories, I-PSS quality of life score and post-void residual urine volume. Secondary outcomes included treatment failure, hospital stay and major complications. Followup was 7.5 months after randomization. Intent to treat analysis was done using analysis of covariance, proportional odds models and the Newman-Keuls multiple comparisons procedure. RESULTS: Of symptomatic patients 117, 117 and 106 were randomized to receive laser therapy, transurethral prostatic resection and conservative management, respectively. Baseline characteristics were similar. All primary outcomes indicated that transurethral prostatic resection and laser therapy were superior to conservative management, and resection was superior to laser therapy. As measured by combined improved symptoms and maximum urinary flow, a successful outcome was achieved in 81%, 67% and 15% of men who underwent transurethral prostatic resection, laser therapy and conservative management, respectively. Hospital stay was significantly shorter and complications fewer for laser therapy than for resection but catheters were in place significantly longer. Men treated conservatively did not have deterioration or treatment failure. CONCLUSIONS: Laser therapy and transurethral prostatic resection are effective for decreasing lower urinary tract symptoms and post-void residual urine volume as well as improving quality of life and maximum urinary flow in the short term in men presenting with moderate to severe symptoms. Transurethral prostatic resection is superior to laser therapy in terms of effectiveness but some patients may elect laser therapy due to the shorter hospital stay and lower risk of complications. Conservative management may be acceptable and safe in men with lower urinary tract symptoms since we observed no marked deterioration in the short term. PMID- 10840428 TI - Bacteremia and bacteriuria after transrectal ultrasound guided prostate biopsy. AB - PURPOSE: We determined the incidence and predisposing factors of bacteremia and bacteriuria after prostate biopsy with specific emphasis on the value of a pre biopsy enema. MATERIALS AND METHODS: We randomized 50 men undergoing ultrasound and biopsy to rule out prostate cancer to receive a preoperative (25) or no (25) enema. Preoperatively urine was obtained for culture, and questionnaires regarding urological history and voiding symptoms were completed. The initial prostate biopsy, biopsy needle, and postoperative urine and blood specimens were cultured. The following day a symptom questionnaire was completed. RESULTS: Bacterial growth in post-procedure cultures did not correlate with the number of biopsies, prostate specific antigen, obstructive voiding symptoms, prostate volume, cancer or post-biopsy hematuria. Bacteriuria was noted in 44% of the cases and bacteremia was present in 16% of the patients, of whom 87.5% did not receive an enema (p = 0.0003). Only 1 patient had chills and fever greater than 37.5C, requiring additional antibiotics. On the followup questionnaire 12% of patients described dysuria, including 84% with bacteriuria after biopsy. CONCLUSIONS: Bacteremia and bacteriuria after multiple biopsies are common but usually asymptomatic. Bacteria is apparently introduced into the urine and/or blood from the rectum via the biopsy needle, which may be minimized by a pre biopsy enema. Dysuria or a history of urinary tract infection did not predict problems after biopsy. PMID- 10840429 TI - Patterns of treatment of patients with prostate cancer initially managed with surveillance: results from The CaPSURE database. Cancer of the Prostate Strategic Urological Research Endeavor. AB - PURPOSE: We determined the demographic and clinical profile of men who elect surveillance as the initial management of prostate cancer as well as the incidence and predictors of secondary treatment of these patients. MATERIALS AND METHODS: The Cancer of the Prostate Strategic Urological Research Endeavor (CaPSURE) is a national disease registry of patients with various stages and treatments of prostate cancer. Using this database of 4,458 men we identified 329 (8.2%) who elected surveillance as the initial management of prostate cancer. Patients choosing watchful waiting were compared to other CaPSURE participants using the chi-square test. The likelihood of treatment initiation in the watchful waiting group was calculated using the Kaplan-Meier method. After adjusting for patient age, race, prostate specific antigen (PSA) at diagnosis, clinical T stage and total Gleason score the Cox proportional hazards regression model was used to determine significant predictors of treatment initiation. RESULTS: Compared with others in the database, patients on watchful waiting were more likely to be 75 years old or older (51% versus 16%, p <0.001), white (93% versus 85%, p <0.001), and have lower serum PSA (p <0.001), organ confined disease (97% versus 88%, p <0.001) and a total Gleason score of 7 or less (97% versus 88%, p <0.001). In the watchful waiting group there was a 52% likelihood of treatment initiation within 5 years of the diagnosis. Significant predictors of secondary treatment were age younger than 65 years and elevated serum PSA at diagnosis. Neither race, extraprostatic stage cT3 disease nor higher total Gleason score was a significant predictor of treatment. CONCLUSIONS: Men who elect initial watchful waiting for prostate cancer tend to be older, have lower serum PSA and more favorable disease characteristics than those who seek treatment. PSA at diagnosis is the dominant factor for predicting secondary treatment. PMID- 10840430 TI - Ability of the 1992 and 1997 American Joint Committee on Cancer staging systems for prostate cancer to predict progression-free survival after radical prostatectomy for stage T2 disease. AB - PURPOSE: In a revised 1997 edition the American Joint Committee on Cancer (AJCC) staging for prostate cancer reduced the subdivision of T2 disease from 3 (T2a, T2b and T2c) to 2 substages by combining single lobe disease (T2a and T2b) into a single stage, now termed T2a. We retrospectively reviewed a large series of men with clinically organ confined prostate cancer who underwent surgery by a single surgeon to determine how this change affected the ability of the staging system to predict outcome. MATERIALS AND METHODS: Of 2, 293 cases treated with radical prostatectomy from 1982 to 1998, 1, 314 were clinically staged to have T2 disease. We analyzed the freedom from biochemical prostate specific antigen (PSA) progression after radical prostatectomy according to the 1992 and 1997 AJCC staging criteria. RESULTS: Overall the actuarial 5 and 10-year biochemical progression-free survival rates for the whole group were 82% and 71%, respectively. The actuarial recurrence-free rate was significantly different for patients with 1992 AJCC T2a (T2a92) versus T2b (T2b92) disease (p <0.0001) but similar for those with 1992 T2b versus T2c (T2c92) disease (p = 0.1165). The actuarial recurrence-free rate was significantly different for patients with 1997 AJCC T2a (T2a97) versus T2b (T2b97) disease (p = 0.0001). However, the overall recurrence-free rate was lower in the T2a97 group compared with the T2a92 group. CONCLUSIONS: The 1992 AJCC staging of T2 prostate cancer yielded a significant difference in the outcome of patients with single lobe disease (T2a92 and T2b92), which is obscured by the 1997 staging criteria. Therefore, we believe that the next revision of the AJCC staging system should revert back to the 1992 AJCC system of subdividing a single lobe disease category into T2a and T2b. Alternatively the committee may want to merge T2b92 and T2c92 stages into a new staging entity, since there was no statistical difference in the outcome of these 2 groups. PMID- 10840431 TI - Management of a positive surgical margin after radical prostatectomy: decision analysis. AB - PURPOSE: We created and tested a decision analysis model to help determine the preferred management of a positive surgical margin(s) after radical prostatectomy. MATERIALS AND METHODS: We constructed a decision tree modeling surveillance versus immediate prophylactic adjuvant radiation in patients with a positive surgical margin(s) after radical prostatectomy. Literature and institution based estimates were determined for certain factors, including the probability of undetectable prostate specific antigen (PSA) in patients followed expectantly postoperatively and those treated with immediate adjuvant radiotherapy, complications of radiotherapy after prostatectomy and probability of undetectable PSA in those treated with therapeutic radiation for detectable PSA postoperatively. A panel of experts assigned utilities to the various outcomes. Sensitivity analysis was performed to determine threshold values required to change the model outcome. RESULTS: Using average probability estimates from a literature review the decision model recommended initial surveillance. Sensitivity analysis demonstrated that the model depended on the probability of disease recurrence in men followed expectantly after surgery as well as the efficacy of therapeutic radiation. We tested the decision model again for patient groups based on tumor grade, pathological stage, preoperative PSA and number of positive margins. The model recommended initial radiation for patients with low to intermediate grade disease, no evidence of seminal vesicle invasion and multiple positive margins. CONCLUSIONS: The results of our decision analysis imply that immediate radiation may be appropriate for patients with a positive surgical margin(s) and a high likelihood of recurrent local rather than distant disease. This model may be useful to physicians and patients who use individual probability estimates and utility values to determine the preferred course of management after surgery. PMID- 10840432 TI - Long-term hazard of progression after radical prostatectomy for clinically localized prostate cancer: continued risk of biochemical failure after 5 years. AB - PURPOSE: Cure from malignancy is commonly defined as a disease-free state lasting 5 years after treatment. We analyzed clinical and biochemical progression rates after radical prostatectomy for men with clinically localized prostate cancer with particular attention to recurrence beyond 5 years. Annual hazard rates of progression were calculated to determine the probability of recurrence at specific intervals following surgery. MATERIALS AND METHODS: The records of 2,782 men with clinically localized prostate cancer (cT1-T2) undergoing radical prostatectomy between 1987 and 1993 were reviewed. All patients were treated in the prostate specific antigen (PSA) era so that serial followup PSA values were available from the time of surgery. Analysis was limited to patients who did not receive adjuvant treatment within 90 days of radical prostatectomy. Disease progression was defined as documented local recurrence, systemic progression and/or PSA 0.4 ng./ml. or greater. Lymph node positive cases were eliminated from analysis since almost all received adjuvant hormonal therapy. Annual hazard rates for progression were calculated using the formula: [No. events / No. patients at risk] x 100. Progression-free survival probabilities were determined using the Kaplan-Meier method. RESULTS: Pathological stage was pT2a-b, N0 (68%), pT3a, N0 (21%) and pT3b, N0 (11%). Biochemical progression-free survival at 5 and 10 years was 76% and 59%, respectively, for the entire study population while those with pathologically organ confined (pT2, N0) cancers had progression-free survival rates of 82% and 68% at 5 and 10 years, respectively. A total of 819 patients (29%) eventually had disease progression, including 160 (6%) with progression after 5 years. Annual hazard rates were highest during the first 2 years after radical prostatectomy for the entire population. Patients with adverse prognostic features (pT3b, PSA 10 ng./ml. or greater, Gleason score 8-10 and nondiploid cancers) had high initial hazard rates that decreased with time to lower levels. Those with pathologically organ confined cancer had low but constant hazard rates throughout followup. CONCLUSIONS: Although progression after radical prostatectomy usually occurs early, reflecting the impact of clinical under staging, a significant number of men, including those with organ confined cancers, will continue to have disease progression after 5 years. Patients undergoing radical prostatectomy should be subjected to long-term followup to allow the option of early intervention should progression occur. PMID- 10840433 TI - Urologists at work. Method permitting easier resection of bladder tumors in the anterior midline. PMID- 10840435 TI - Detrusor pressure uroflowmetry studies in women: effect of a 7Fr transurethral catheter. AB - PURPOSE: We evaluated whether a 7Fr transurethral catheter affects urinary flow in women undergoing pressure flow studies for voiding symptoms. MATERIALS AND METHODS: We reviewed a urodynamic database of 600 consecutive women referred for the evaluation of voiding symptoms. Before urodynamics all patients voided privately using a standard toilet and free flow was recorded. Urodynamics were performed using a 7Fr double lumen transurethral catheter. At functional bladder capacity patients were asked to void in the sitting position and pressure flow studies were performed. All uroflowmetry tracings were inspected and analyzed manually. Only patients who voided similar volumes varying by less than 20% on the free and pressure flow studies were assessed. Free and pressure flow parameters were compared according to voided volume category, main urodynamic diagnosis, uroflowmetry pattern and pre-void bladder volume. RESULTS: A similar volume was voided on the free and pressure flow studies of 100 women. In each voided volume category and urodynamic diagnosis pressure flow parameters were significantly different from the equivalent free flow parameters in all but 4 cases. Specifically the maximum flow rate was significantly less and flow time was significantly longer on pressure versus free flow studies (each p <0.01). An intermittent flow pattern was more common on pressure than in free flow measurements (43% versus 9%). CONCLUSIONS: A 7Fr transurethral catheter may adversely affect uroflowmetry parameters in women undergoing pressure flow studies for lower urinary tract symptoms. This finding may have further clinical implications regarding the interpretation of these parameters as well as establishment of an accurate diagnosis. PMID- 10840434 TI - Penile straightening with crural graft of the corpus cavernosum. AB - PURPOSE: We present a surgical alternative to penile straightening in Peyronie's disease that avoids penile shortening by using tissue from the crural segment of the corpora cavernosa as a graft. MATERIALS AND METHODS: We treated 7 patients with stable Peyronie's disease and satisfactory penile rigidity but significant curvature that precluded intercourse with this technique. An incision is made in the fibrous area and after stretching the gap is covered with a graft removed from the crural segment of the corpora cavernosa. RESULTS: In 6 of the 7 men straightening was satisfactory but in 1 it was not complete. No patient reported worse penile rigidity postoperatively. Donor site healing was uneventful. CONCLUSIONS: Use of tissue from the corpus cavernosum to correct penile curvature is effective. However, our results are preliminary, and long-term effectiveness and safety must be confirmed in a larger number of cases. PMID- 10840436 TI - Urodynamics in spinal cord injured patients walking with reciprocating gait orthosis. AB - PURPOSE: We evaluated the effects of walking with reciprocating gait orthosis on bladder function and upper urinary tract status in spinal cord injured patients. MATERIALS AND METHODS: Seven patients using reciprocating gait orthosis underwent urodynamics before and during walking. Abdominal ultrasound was performed before and after urodynamics. RESULTS: Baseline urodynamics showed detrusor areflexia in 3 patients and hyperreflexia in 4. During walking urodynamics revealed hyperreflexia in 6 patients with worse uninhibited detrusor contraction amplitude (38.2 +/- 41.2 versus 73.7 +/- 44.4 cm. water, p = 0.006) and duration (2.1 +/- 2.1 versus 6.8 +/- 5.3 minutes, p = 0.023). After walking ultrasound demonstrated bilateral pyelectasia in 2 patients. CONCLUSIONS: Spinal cord injured patients using reciprocating gait orthosis have worse urodynamic measurements and upper urinary tract disorders during walking. They require close urological followup. PMID- 10840437 TI - Stone granuloma causes ureteropelvic junction obstruction after percutaneous nephrolithotomy and antegrade endopyelotomy. PMID- 10840439 TI - Use of the nonrefluxing native ureter for urinary diversion. PMID- 10840438 TI - Upper urinary tract cholesteatoma misdiagnosed as a ureteral tumor. PMID- 10840440 TI - A case of urethrocutaneous fistula with the koro syndrome. PMID- 10840441 TI - Development of urethrovaginal fistula following periurethral collagen injection. PMID- 10840442 TI - Pure seminoma in a male phenotype 46,XX true hermaphrodite. PMID- 10840444 TI - Re: Complications of retrograde balloon cautery endopyelotomy. PMID- 10840443 TI - Resolution of the chronic pelvic pain syndrome after renal transplantation. PMID- 10840445 TI - Vaginal calculi secondary to partial vaginal outlet obstruction in pediatric patients. PMID- 10840446 TI - Standardized ultrasound method for assessing detrusor muscle thickness in children. AB - PURPOSE: We standardized transabdominal ultrasound measurement of bladder wall thickness in children and evaluated its reliability. MATERIALS AND METHODS: Using ultrasonography we measured the thickness of the low echogenic layer of the ventral and dorsal bladder walls at increasing degrees of bladder fullness in the setting of voiding cystourethrography. For assessing intra-observer variability an observer blinded to the numerical values of the measurements studied 38 patients 0.2 to 13.7 years old (median age 1. 5). Measurements were performed as pairs of replicates at well-defined sites with reference to the urachal remnant, and rectal impression and/or contour. For assessing interobserver variability 3 independent observers measured the same child. Eight children 0.3 to 10.5 years old (median age 6.4) were evaluated at 2 degrees of bladder fullness per child. RESULTS: The thickness of the low echogenic layer of the ventral and dorsal walls was 0.4 to 2.9 (median 0.9) and 0.4 to 2.8 mm. (median 1.1). The dorsal wall was slightly thicker than the ventral wall. The intra-observer and interobserver variability of measurements (standard deviation) was 0. 2 mm. for each wall part. There was a small systematic difference among observers. CONCLUSIONS: It is possible to determine the thickness of the low echogenic layer of the bladder wall with a systematic and anatomically defined method of acceptably reliable measurements. The ventral and dorsal walls should continue to be measured until more is known about their pathological appearance. PMID- 10840447 TI - Urodynamic findings in boys with posterior urethral valves after treatment with primary valve ablation or vesicostomy and delayed ablation. AB - PURPOSE: Primary valve ablation and temporary vesicostomy with delayed valve ablation are alternative initial management procedures in neonates and infants with posterior urethral valves. To investigate whether initial vesicostomy followed by delayed valve ablation and simultaneous vesicostomy closure may lead to more alterations in bladder function than primary valve ablation only we retrospectively compared postoperative urodynamic findings in 2 small groups of patients. MATERIALS AND METHODS: From 1980 to 1990, 15 male infants 19 days to 34 months old with posterior urethral valves were treated with 1 of 2 initial surgical approaches, including valve ablation only in 8 (group 1), and primary vesicostomy and delayed valve ablation associated with concomitant vesicostomy closure in 7 (group 2). Mean age at valve ablation and vesicostomy in groups 1 and 2 was 10.8 +/- 11.2 months (range 1 to 35) and 55.4 +/- 43.3 days (range 19 to 151), respectively. Average duration of vesicostomy diversion was 33.6 +/- 18.8 months (range 14 to 70). All patients underwent conventional urodynamics postoperatively using normal saline at room temperature. In groups 1 and 2 mean age at followup was 11.5 +/- 6.6 (range 5 to 16.2) and 9. 4 +/- 3.1 (range 4.10 to 14) years, respectively. Controls comprised 46 age matched males who underwent urodynamics using similar methodology. RESULTS: Postoperative urodynamic assessment of maximum cystometric bladder capacity and the incidence of detrusor instability in each treatment group were not statistically different. In group 1 bladder capacity was significantly higher than that in controls (p <0.0001). In group 2 mean end filling detrusor pressure was increased compared with that in group 1 (29 cm. water, range 15 to 60 versus 8, range 4 to 21). Compliance was significantly lower in group 2 than in group 1 (p <0.0005). Analysis of detrusor voiding pressure at maximum flow was not significantly different in the 2 groups. We noted detrusor under activity in 1 group 1 and 2 group 2 cases. In these patients post-void residual urine volume was 8% to 66% of cystometric bladder capacity. However, only 1 of these 3 patients who required augmentation cystoplasty needed intermittent catheterization. Urodynamic patterns of outflow obstruction developed in 1 patient in each group, including urethral stricture and bladder neck obstruction. At followup we observed no difference in renal function impairment in the 2 groups. CONCLUSIONS: Our retrospective study of rapid filling cystometry suggests that primary valve ablation for posterior urethral valves is associated with a better bladder function outcome than that in patients treated with vesicostomy and delayed valve ablation. Therefore, although cutaneous vesicostomy may be performed as initial management of posterior urethral valves, primary valve ablation is the most effective surgical option in these cases. PMID- 10840449 TI - Posterior urethral valves and the 11th Commandment. PMID- 10840448 TI - Temporary high diversion for posterior urethral valves. AB - PURPOSE: Temporary high diversion for posterior urethral valves remains controversial. Even in the most severe cases some physicians deny the efficacy of this treatment. They assert that high diversion does not change the outcome of kidney function and has an iatrogenic, deleterious effect on the bladder. We believe that these 2 assertions may be inaccurate. MATERIALS AND METHODS: We evaluated 17 of 120 boys with posterior urethral valves who underwent temporary high diversion via Sober-en-T ureterostomy. This procedure immediately decompresses the upper urinary tract and leaves the bladder functional. Mean duration of diversion was 13 months. Bladder function results were good and diversion clearly had no deleterious effect. However, our series was too small to conclude with certainty that renal function improved due to diversion and not to valve resection only. Rapid improvement in creatinine was noted in all cases after diversion. RESULTS: In this study we reviewed opposing opinions. It appears clearly evident that physicians who believe that ureterostomy creates a valve bladder are considering only loop diversion, which temporarily defunctionalizes the bladder. On the contrary, Sober-en-T diversion preserves bladder cycling. In addition, those who report poor kidney function after high diversion fail to mention that this procedure is usually performed only in the most severe cases. CONCLUSIONS: For severe cases of posterior urethral valves one should not hesitate to create temporary high diversion, which may possibly improve renal function. Sober-en-T ureterostomy does not damage the bladder. PMID- 10840450 TI - Preservation of the thin distal urethra in hypospadias repair. AB - PURPOSE: We report our experience with preserving the thin distal urethra lacking corpus spongiosum for the treatment of hypospadias. MATERIALS AND METHODS: From January 1997 to October 1999 we treated primary hypospadias in 77 boys with a mean age of 4 years 10 months. After degloving the penile skin a segment of thin distal urethra lacking corpus spongiosum was noted in 18 patients (23.4%) with a mean age of 3 years 5 months. The thin distal urethra was preserved and incorporated as part of urethroplasty (group 1). The procedure was primarily completed by tubularized incised plate urethroplasty. We also performed tubularized incised plate urethroplasty in 31 boys (40.3%) with a mean age of 5 years 2 months who had normal coverage of the corpus spongiosum of a hypospadiac urethra (group 2). We compared the results of treatment in these 2 groups. The remaining 28 patients (36.4%) treated with other methods were excluded from study. RESULTS: In group 1 the distance from the original meatus to the urethra covered by healthy corpus spongiosum was 4 to 20 mm. (mean 8.2). If the thin distal urethra had been excised, the urethral meatus would have been relocated more proximal in these boys. Mean followup in groups 1 and 2 was 9.9 and 7.6 months, respectively. Postoperatively there were 2 (11.1%) urethrocutaneous fistulas in group 1 and 4 (12.9%) in group 2. Tubularized incised plate repair was successful in all 10 cases (100%) of distal hypospadias in group 2, and in 15 of 18 (83.3%) and 17 of 21 (81%) of proximal hypospadias cases in groups 1 and 2, respectively. There was no statistically significant difference in the success rate of hypospadias repair in the groups. CONCLUSIONS: We noted a significantly thin distal urethra in 23.4% of our cases of primary hypospadias. Mean length of the thin distal urethra was 8.2 mm. Preserving the thin distal urethra may simplify the operative procedure without compromising the surgical results of tubularized incised plate urethroplasty. PMID- 10840451 TI - The application of magnetic resonance imaging for the preoperative localization of nonpalpable testis in obese children: an alternative to laparoscopy. PMID- 10840452 TI - Single scrotal incision orchiopexy for the palpable undescended testicle. AB - PURPOSE: We prospectively evaluated the Bianchi single high scrotal incision technique for orchiopexy in boys with palpable undescended testis distal to the external inguinal ring. MATERIALS AND METHODS: A total of 60 Bianchi orchiopexies were performed in 48 patients. We identified a group with primary undescended and secondary ascended testis, and a trapped testicle associated with previous inguinal surgery, respectively. Testicular position and size were assessed at 6 weeks of followup. RESULTS: We evaluated group 1-34 patients with bilateral (8), right (17) and left (9) primary undescended testis, group 2-3 with secondary ascended testis and group 3-11 with bilateral (4), right (4) and left (3) trapped testis. Average surgical time was 15, 35 and 35 minutes, respectively. Four patients required conversion to a traditional inguinal incision to achieve adequate cord length. All but 2 patients had a palpable testicle of acceptable size compared with the contralateral mate in the dependent scrotum at the end of the operation and 6 weeks postoperatively. Those returning for 1-year followup had identical findings. In groups 1 and 2 a single testicle normal to palpation resided in a high scrotal position and 1 testis required secondary orchiectomy for infarction, respectively. Scrotal hematoma resolved spontaneously without injury to the testis in 1 case. No hernias were evident. CONCLUSIONS: A single high scrotal incision for palpable primary, secondary ascended or even trapped testis is well tolerated, cosmetically pleasing and associated with a short operative time. The complication rate is acceptably low. PMID- 10840453 TI - Orchiopexy using 2 mm. laparoscopic instruments: 2 techniques for delivering the testis into the scrotum. AB - PURPOSE: We describe modified techniques of laparoscopic orchiopexy using 2 mm. instruments that enable straightforward delivery of the testis into the scrotum. MATERIALS AND METHODS: The preferred technique involves a radially dilating trocar advanced over the 2 mm. grasper placed antegrade through the new internal ring. The other method is similar but Amplatz dilators are used when a radially dilating trocar system is not available. RESULTS: We used these techniques with complete success to deliver 18 testes into the scrotum. CONCLUSIONS: The testis may be safely and effectively delivered to the scrotum using 2 mm. instruments and disposable fascial dilators or a radially dilating trocar system. PMID- 10840454 TI - A study of the etiology of idiopathic calcium urolithiasis in children: hypocitruria is the most important risk factor. AB - PURPOSE: To determine the association of metabolic risk factors with pediatric calcium urolithiasis we compared metabolic evaluation data on children with idiopathic calcium stones and those on healthy children. MATERIALS AND METHODS: Metabolic evaluation was done in 78 calcium stone formers 1 to 15 years old (mean age 7.2) who were free of urinary tract infection, anatomical abnormalities, and metabolic, endocrinological and intestinal disorders, and in 24 healthy children. Evaluation included serum biochemistry, and measurement of daily excretion of urinary calcium, oxalate, urate, phosphorus, citrate and magnesium. RESULTS: Demographic characteristics, serum parameters, and daily excretion of calcium, urate, phosphorus and magnesium did not differ statistically in the 2 groups. However, urinary oxalate was significantly higher and urinary citrate was significantly lower in stone formers than in controls (p = 0.002 and 0.028, respectively). Hypocitruria and hyperoxaluria were 4.3 and 3-fold more common in stone formers than in controls, respectively. Multivariate analysis using logistic regression showed that hypocitruria was the only significant risk factor for idiopathic calcium stones (p = 0.008). CONCLUSIONS: Hypocitruria was the most important risk factor in our patients. Hyperoxaluria was also common and accompanied hypocitruria in many stone formers. In contrast to many previous reports, we failed to show that hypercalciuria is an important metabolic defect for idiopathic calcium stones, possibly because our study evaluated a different population. PMID- 10840455 TI - Review article: The molecular era of bladder research. Transgenic mice as experimental tools in the study of outlet obstruction. AB - PURPOSE: To review the crucial role of transgenic mice as experimental tools in the study of outlet obstruction. MATERIALS AND METHODS: We reviewed the literature for studies that have used mice as models for outlet obstruction. RESULTS: The combination of genetic manipulations and cellular physiology defines state-of-the-art experiments that explore the reciprocal mesenchymal-epithelial interactions that regulate bladder cell mechanisms. CONCLUSIONS: The use of transgenic mice in bladder research has provided important data with respect to the molecular signals that drive bladder development, homeostasis, and the response to injury. PMID- 10840456 TI - Experimental diabetes-induced regression of the rat prostate is associated with an increased expression of transforming growth factor-beta. AB - PURPOSE: Transforming growth factor-beta (TGF-beta), a potent inhibitor of cell growth, plays an important role in the androgen-dependent processes of the prostate through a complex network of growth factors. TGF-beta expression in the prostate is under negative regulatory control of androgen. As experimental diabetes causes a regression of the prostate and decrease in serum testosterone levels in rats, we examined TGF-beta alterations at the mRNA and protein levels in the diabetic rat prostate. MATERIALS AND METHODS: The expression of TGF-beta1 and TGF-beta2 and their respective mRNAs in prostates from streptozotocin (STZ) induced diabetic, insulin-treated diabetic and age-matched control rats were investigated, using relative multiplex RT-PCR, semi-quantitative Western blotting, and immunohistochemistry. RESULTS: Induction of diabetes caused a significant reduction in prostatic weight and in serum testosterone levels in rats. Both mRNA and protein levels of TGF-beta1, and mRNA level of TGF-beta2 were up-regulated in the diabetic rat prostate. Insulin-treatment normalized changes observed in prostatic weight and serum testosterone levels, and reversed the alterations in the TGF-beta1 and TGF-beta2 expression at the gene transcript and protein levels to control levels. Immunohistochemical studies demonstrated that TGF-beta1 is localized to prostatic stromal cells, whereas TGF-beta2 is located in both epithelial and stromal cells. CONCLUSION: These results suggest that TGF beta1 and TGF-beta2 may be involved in the diabetes-induced regression of the prostate gland. PMID- 10840457 TI - Angiotensin II type 1 receptor antagonist (losartan) down-regulates transforming growth factor-beta in experimental acute pyelonephritis. AB - PURPOSE: To study the effect of an angiotensin II type 1 receptor antagonist, losartan, on cytokine expression, kidney growth and renal scarring in experimental acute pyelonephritis. MATERIALS AND METHODS: Female Bki NMRI mice, 8 weeks old were infected with E. coli CFT 073 via the urethra. Mice were divided into four groups; either left untreated; or treated with NaCl 0.9%; or an angiotensin II type 1 receptor antagonist, losartan, in doses of 1 mg. or 40 mg. /kg. body weight. The treatment was given daily i.p. for 48 hours, 3 weeks or 8 weeks respectively. Kidneys were weighed and sectioned for histo-pathology and in situ hybridization for mRNA of IL-1beta, TNF-alpha, IL-4, IL-6, IL-10, IL-12, TGF beta and IFN-gamma. Homogenized kidneys were used for EIA of TGF-beta and bacterial growth. RESULTS: The mRNA expression of the studied cytokines generally peaked at 48 hours in all four groups. In animals treated with losartan, kidney TGF-beta, IFN-gamma and IL-6 decreased significantly at 3 and 8 weeks as compared with controls, untreated or those treated with NaCl, (p <0.005 respectively). Infection was associated with a declining kidney weight, also in the presence of losartan. A 50% reduction of the spread of renal scarring was observed in the losartan treated group, but this did however not reach significance. The proportion of kidneys showing bacterial growth was not influenced by losartan although in these kidneys the mean bacterial counts at 3 weeks were significantly higher in the losartan treated mice (p <0.006). CONCLUSIONS: Losartan is associated with downregulation of TGF-beta, IFN-gamma and IL-6 and may, in combination with antimicrobial therapy, reduce the risk of cortical renal scarring in recurrent acute pyelonephritis in infants. PMID- 10840458 TI - Loss of heterozygosity at 12P12-13 in primary and metastatic prostate adenocarcinoma. AB - PURPOSE: Our laboratory has recently identified a 1 to 2 Mb homozygous deletion at 12p12-13 in a prostate cancer specimen and determined that the p27/kip1 gene lies within the deletion. While immunohistochemical analysis has implicated p27/kip1 in prostate carcinoma, no previous studies had identified genetic abnormalities at this locus. Here, we examined primary and metastatic prostate tumors to determine if allelic loss occurs at this locus in localized disease and if it increases the risk of metastatic, high stage or high-grade disease. MATERIALS AND METHODS: DNA was extracted from prostate tumors and normal tissue of 99 patients. 60 tumors were primary, 20 were metastatic pelvic lymph nodes, and 19 were distant metastases. Multiple metastases were analyzed from 11 of 19 patients with metastatic disease. Polymorphic markers spanning our region of interest were PCR amplified from tumor and normal DNA. PCR products were then scored for allelic loss. RESULTS: Loss of heterozygosity (LOH) was identified in 14/60 (23%) primary tumors, 6/20 (30%) lymph node metastasis, and 9/19 (47%) distant metastases. The difference between primary and distant metastatic disease was statistically significant (p = 0.045, Fisher's exact test). The pattern of LOH was identical in all metastatic sites obtained from individual patients, indicating that genetic loss occurred prior to metastasis. Subset analysis of the 60 primary tumors demonstrated no association between LOH and adverse pathological feature [nodal involvement, seminal vesicle invasion, margin positivity, high Gleason score (7-10)]. CONCLUSIONS: Demonstrating that 12p12-13 LOH is a prominent feature of primary prostate tumors and that multiple metastatic foci have an identical LOH pattern, provides evidence that gene inactivation in this region occurs prior to metastasis. In addition, the strong association between LOH and distant metastasis raises the possibility that mutational inactivation of a gene at 12p12-13, possibly p27/kip1, plays a pivotal role in the development of metastatic disease. PMID- 10840459 TI - Role of neurokinin receptors in the behavioral effect of intravesical antigen infusion in guinea pig bladder. AB - PURPOSE: To characterize a guinea pig behavior model of bladder pain due to intravesical antigen infusion and to determine the role of neurokinin receptor subtypes in mediating this behavior. MATERIALS AND METHODS: The influence of subtype-selective neurokinin receptor antagonists on increased abdominal licking behavior in response to intravesical antigen infusion in guinea pigs immunized with ovalbumin (OA) was determined. RESULTS: Intravesical OA infusion for 30 minutes induced a significantly greater frequency (about 3-fold) of abdominal licking behavior than during either the 30 minutes pre-challenge or post challenge saline infusions. Treatment with IP capsaicin 7 to 10 days before OA challenge abolished the intravesical antigen-induced behavior. IP injection of the NK1 receptor antagonist CP 99994 (10 mg./kg. or 30 mg./kg.), 30 minutes pretreatment, inhibited the increase in the average number of abdominal licks during antigen infusion. The 30 mg./kg., but not the 10 mg./kg. dose increased the percent of animals showing antinociceptive activity (defined as 4 or less abdominal licks during the antigen infusion). The NK2 receptor antagonist SR 48968 reduced the antigen-induced abdominal licking behavior at IP doses of 3 and 10 mg./kg. but was ineffective at 1 mg./kg. The NK3 receptor antagonist SB 235375 (30 mg./kg., IP) did not reduce this behavior. CONCLUSIONS: These results suggest a role for activation of NK1 and NK2, but not NK3 receptors, by tachykinins released from capsaicin-sensitive nerves, in the increased abdominal licking behavior response of guinea pigs to intravesical antigen infusion. PMID- 10840460 TI - Cyclophosphamide-induced cystitis in freely-moving conscious rats: behavioral approach to a new model of visceral pain. AB - PURPOSE: To develop a model of visceral pain in rats using a behavioral approach. Cyclophosphamide (CP), an antitumoral agent known to produce toxic effects on the bladder wall through its main toxic metabolite acrolein, was used to induce cystitis. MATERIALS AND METHODS: CP was administered at doses of 50, 100 and 200 mg./kg. i.p. to male rats, and their behavior observed and scored. The effects of morphine (0.5 to 4 mg./kg. i.v.) on CP-induced behavioral modifications were tested administered alone and after naloxone (1 mg./kg. s.c.). In addition, 90 minutes after CP injection, that is, at the time of administration of morphine, the bladder was removed in some rats for histological examination. Finally, to show that the bladder is essential for the CP-induced behavioral modifications, female rats also received CP at doses of 200 mg./kg. i.p. and of 20 mg. by the intravesical route, and acrolein at doses of 0.5 mg. by the intravesical route and of 5 mg./kg. i.v. RESULTS: CP dose-relatedly induced marked behavioral modifications in male rats: breathing rate decrease, closing of the eyes and occurrence of specific postures. Morphine dose-dependently reversed these behavioral disorders. A dose of 0.5 mg./kg. produced a reduction of almost 50% of the behavioral score induced by CP 200 mg./kg. This effect was completely prevented by pretreatment with naloxone. At the time of administration of morphine, histological modifications of the bladder wall, such as chorionic and muscle layer edema, were observed. In female rats, CP 200 mg./kg. i.p. produced the same marked behavioral modifications as those observed in male rats. Administered at the dose of 20 mg. intravesically, CP did not produce any behavioral effects, whereas acrolein at 0.5 mg. intravesically induced behavioral modifications identical to those under CP 200 mg./kg. i.p., with the same maximal levels. Conversely, acrolein 5 mg./kg. i.v. did not produce any behavioral effects at all. CONCLUSIONS: Overall, these results indicate that this experimental model of CP-induced cystitis may be an interesting new behavioral model of inflammatory visceral pain, allowing a better understanding of these painful syndromes and thus a better therapeutic approach to them. PMID- 10840461 TI - Polymorphisms of N-acetyltransferase 2, glutathione S-transferase mu and theta genes as risk factors of bladder cancer in relation to asthma and tuberculosis. AB - PURPOSE: We investigated whether the polymorphisms of N-acetyltransferase 2 (NAT2), glutathione S-transferase-mu (GSTM1) and glutathione S-transferase-theta (GSTT1) genes were risk factors of bladder cancer among Korean people in relation to other risk factors. MATERIALS AND METHODS: In this case-control investigation of 113 patients with primary bladder cancer and 221 control subjects, we compared the association of bladder cancer with genetic polymorphisms of NAT2, GSTM1 and GSTT1, demographic characteristics, smoking status, and medical histories in a molecular epidemiological way. RESULTS: The risk of bladder cancer was significantly increased in patients with a medical history of tuberculosis (OR 3.61, 95% CI 1.57 to 8.26) and bronchial asthma (OR 4.15, 95% CI 1.61 to 10.75), while smoking history turned out to be insignificant. GSTM1 null genotype was a significant risk factor of bladder cancer (OR 1.81, 95% CI 1.12 to 2.93). On the other hand, slow acetylator and GSTT1 genotypes were insignificant. Also, we could not find any association between GSTM1, GSTT1, slow acetylator genotypes and bladder cancer risk among smokers. The rapid acetylator, GSTM1 null and GSTT1 null genotypes with a medical history of asthma or tuberculosis combinations were significant risk factors in Korean subjects. CONCLUSIONS: Among Korean subjects, GSTM1 null genotype was a significant risk factor for bladder cancer. The reason why bronchial asthma and tuberculosis are risk factors in Korean subjects is yet unknown, but a variety of factors, including enzyme activities for detoxification, medication for these diseases and immunological background might be involved. PMID- 10840462 TI - IL-1beta and TNF-alpha in prostatic secretions are indicators in the evaluation of men with chronic prostatitis. AB - PURPOSE: Chronic Prostatitis, or Chronic Pelvic Pain Syndrome [CPPS], is a common disorder characterized by pelvic pain and varying degrees of inflammation in expressed prostatic secretions (EPS). In search of markers to more clearly define CPPS, we compared proinflammatory cytokines interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) levels in EPS from men with CPPS, to healthy men and men with Benign Prostatic Hyperplasia (BPH). METHODS: 78 men: controls (n = 16), BPH (n = 14), CPPS IIIA [>/=10 white blood cells per high power field (WBC/hpf) in EPS] (n = 18), CPPS IIIB [<10 WBC/hpf in EPS] (n = 20), and asymptomatic inflammatory prostatitis (AIP) (n = 10) were evaluated for EPS WBC, and IL-1beta and TNF-alpha by ELISA. RESULTS: IL-1beta and TNF-alpha levels in EPS were usually detectable in men with CPPS IIIA (89% and 45%, respectively) or AIP (90%; 100%), but less often in controls (31%; 17%), BPH (57%; 15%), and CPPS IIIB (35%; 15%) respectively. IL-1beta and TNF-alpha levels were higher in CPPS IIIA versus CPPS IIIB, and in AIP versus controls or BPH (p's <0.001). Cut points for IL-1beta and TNF-alpha discriminated AIP from controls (predictive values = 94% and 83%, respectively) and CPPS IIIA from CPPS IIIB (predictive values 84% and 100%). Overall, there was a correlation between IL-1beta and TNF alpha (p <0.003), but no correlation between WBC and IL-1beta (p <0.1) or TNF alpha (p <0.50). CONCLUSIONS: Cytokines are frequently present and elevated in the EPS from men with CPPS IIIA and AIP and provide a novel means for identification, characterization and potential management of men with CPPS that differs from traditional methods based on WBC. PMID- 10840463 TI - Urea modifies the permeability of the mammalian urothelium. AB - PURPOSE: This study investigated the effects of mucosal (urine side) and serosal (blood side) urea on the permeability properties of the in vitro mammalian urinary bladder epithelium. MATERIALS AND METHODS: The permeability properties of the rabbit urinary bladder epithelium were studied in modified Ussing chambers using electrophysiological techniques. RESULTS: Addition of two molar urea to the mucosal solution did not cause a significant change in the short circuit current (Isc, a measure of the ion transport capacity of the epithelium), or the transepithelial conductance (Gt, a measure of the ability of ions to diffuse across the epithelium). In contrast, addition of 0.5 M urea to the serosal solution caused an increase in Gt of approximately 35 microS/cm.2 as well as an increase in Isc over a 5 minute period. The site of the conductance increase by short-term serosal urea was at the apical membrane and not at the tight junctions. The urea-induced conductance completely reversed upon removal of urea, was non-selective, and the magnitude was voltage dependent. Long term serosal urea (greater than 30 minutes) resulted in an irreversible increase in transepithelial conductance. Mucosal urea altered the time course but not the magnitude of the serosal urea-induced conductance. CONCLUSIONS: The ion permeability of the mammalian urinary bladder is increased by serosal urea. At short times the increase is at the apical membrane, while at long times the increase is at the tight junctions. The presence of mucosal urea slows the loss of urothelial barrier function caused by serosal urea. PMID- 10840464 TI - Free radical scavengers, catalase and superoxide dismutase provide protection from oxalate-associated injury to LLC-PK1 and MDCK cells. AB - PURPOSE: Current studies have provided evidence that exposure of renal epithelial cells to oxalate and calcium oxalate crystals induces lipid peroxidation and injures the cells. Since oxidant/antioxidant balance is likely to play a critical role, we determined the effect of antioxidant scavengers on production of free radicals and injury to LLC-PK1 and MDCK cells from exposure to oxalate (Ox) or Ox + calcium oxalate monohydrate (COM) crystals. MATERIALS AND METHODS: LLC-PK1 and MDCK cells were grown in monolayers and exposed to 1.0 mmol. Ox or 1.0 mmol. Ox + 500 microg. /ml. COM crystals for 120 or 240 minutes. We measured the release of lactate dehydrogenase (LDH) as a marker for cell injury and malondialdehyde (MDA) as a marker of lipid peroxidation. Superoxide and hydroxyl radicals were measured in the presence or absence of 400 U/ml. catalase, or superoxide dismutase (SOD). RESULTS: Exposure of LLC-PK1 cells to Ox resulted in a significant increase in MDA and release of LDH, which was further elevated when COM crystals were added. MDCK cells responded similarly to both challenges, but showed significantly less impact when compared with LLC-PK1 cells. Both treatments were associated with significant increase in the generation of hydroxyl and superoxide radicals by both cell types. In both cell lines, the addition of catalase or SOD significantly reduced the increase of MDA and release of LDH. CONCLUSIONS: Results of the present study indicate that both Ox and COM crystals are injurious to renal epithelial cells and the injury is associated with generation of free radicals. Cells of proximal tubular origin are more susceptible than those of distal tubules and collecting ducts. Free radical scavengers, catalase and SOD provide significant protection. PMID- 10840466 TI - Re: Ileal neobladder and local recurrence of bladder cancer: patterns of failure and impact on function in men. PMID- 10840467 TI - Re: Editorial: transrectal ultrasound guided prostate biopsy-defining a new standard. PMID- 10840465 TI - Changes in urine macromolecular composition during processing. AB - PURPOSE: To determine the urinary crystallization inhibitory activity, urine is generally centrifuged and/or filtered. These preparative procedures may result in a total or partial removal of many macromolecular constituents implicated in crystallization. The main purpose of this study was to investigate the changes in urinary macromolecular composition following centrifugation and filtration. MATERIALS AND METHODS: Twenty-four hour urine samples were collected from human volunteers. Each was divided into 4 aliquots; one was filtered, the other was centrifuged, another was centrifuged and filtered. The control sample was neither filtered nor centrifuged. Total protein and lipid contents of each sample were determined. Proteins were analyzed using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Western blot analysis was performed using antibodies against osteopontin (OPN), prothrombin (PT) related proteins, inter alpha-inhibitor (IalphaI) related proteins, Tamm-Horsfall protein (THP), and albumin (ALB). The effect of processing on incorporation of urinary proteins in crystal matrices was also examined. Calcium oxalate crystals were produced in processed and unprocessed urine samples by the addition of sodium oxalate. Crystals were harvested, de-mineralized and their proteins analyzed by SDS-PAGE and Western blotting. RESULTS: Processing reduced the amounts of both proteins and lipids in the urine. Previously we identified phospholipids in the matrix of calcium oxalate crystals as well as the filtrate and retentate removed during filtration and centrifugation. Phospholipids have a high affinity for calcium containing crystals. In the case of proteins, those with high molecular weights appeared to be clearly affected by filtration and centrifugation. Processing also appeared to influence the incorporation of proteins in the crystals. The matrix of crystals produced in processed urine contained less THP than those made in unprocessed urine, apparently a result of the loss of this higher molecular weight protein during processing. Incorporation of PT-related proteins, particularly fragment 1, was increased. CONCLUSIONS: We propose that selective inclusion of macromolecules is a result of an increase in available binding sites on crystal surfaces because of the removal of certain calcium binding substances such as phospholipids and proteins. Removal of larger macromolecules from the milieu may also provide a better access to the crystal surfaces. PMID- 10840469 TI - Re: Erosion of woven polyester pubovaginal sling. PMID- 10840468 TI - Re: Prostate histopathology and the chronic prostatitis/chronic pelvic pain syndrome: a prospective biopsy study. PMID- 10840470 TI - Re: A technique of transrectal ultrasound guided transperineal random prostate biopsy in patients with ulcerative colitis and an ileal pouch. PMID- 10840471 TI - Change of body composition over an eight year period among Japanese university students. AB - This study examined the change of body composition in Japanese university students. Subjects were university students divided into two groups by sex for two different time periods: 67 males and 46 females for 1986-1987 and 47 males and 64 females for 1994-1995. Body height, weight, and underwater weight were measured to estimate the percentage of body fat. The fat mass index (FMI) was applied after adjusting fat mass and the fat-free mass index (FFMI) applied after adjusting fat-free mass for body physique by dividing (body height)2. The mean body mass index (BMI) increased from 1986-1987 to 1994-1995 in males and decreased in females, although there were no statistical differences between two time periods in both sexes. The FMI indicates that in 1994-1995 males had significantly more fat adjusted for body height than in 1987. Females in 1994 1995 had significantly less FFMI than those in 1986 despite FMI similar to that of 1986. Our results thus warn against a trend toward excessive thinness in collegiate females and insufficiency of evaluating body composition using the BMI alone. PMID- 10840472 TI - Body composition, smoking and physical activity in 75-year-old men and women in three Nordic localities with special reference to diagnosed diseases. AB - PURPOSE: To describe and compare anthropometric characteristics among populations of 75-year-olds in three Nordic localities and to investigate possible relationships with chronic heart disease (CHD), chronic lung disease (CLD), diabetes mellitus, arthrosis and life-style factors such as smoking and physical activity. MATERIAL AND METHODS: Anthropometric data were measured in 104 men and 191 women in Jyvaskyla (Finland), 196 men and 209 women in Glostrup (Denmark) and in 127 men and 167 women in Goteborg (Sweden). Variables assessed were body height, body weight, BMI, waist/hip ratio, skinfolds from several sites and percent body fat by bioelectrical impedance. The diagnosis of diseases were made by a physician based on the individual's medical history, drugs and medical examination. Physical activity was recorded by self-rating. RESULTS: Men in Goteborg were taller and had a higher body weight while there was no difference in BMI among the men. Among women, those in Goteborg were tallest, while those in Jyvaskyla had the highest body weight, BMI, percent of body fat and waist/hip ratio. Biceps and triceps skinfolds were highest in men from Goteborg, while in women triceps and subscapular skinfolds were highest in those from Jyvaskyla. CHD was most common in those from Jyvaskyla, and women with CHD had a higher body weight, BMI and lean body mass in all three localities. Lean body mass was lower in men and women with CLD and women with CLD were also shorter, with a lower body weight, BMI, and percent body fat. Diabetes mellitus was associated with a higher body weight, BMI, percent body fat and lean body mass in women, but not in men. Both men and women with arthrosis had a higher BMI, while smoking was only associated with CHD in those from Jyvhskyla. In men with CHD the proportion of persons with low physical activity was higher in all three localities. The physical activity was also lower among men with CLD and diabetes mellitus in Goteborg and Glostrup. CONCLUSION: There were anthropometric differences among 75 year-olds in the three Nordic localities. CHD and CLD were associated with various anthropometric variables. These findings may reflect either cause and effect relationships between diseases and anthropometric characteristics or differences in life-style factors influencing morbidity. PMID- 10840473 TI - "Weight loss preoccupation in aging women": a review. AB - Preoccupation with body weight leading to frequent dieting has been found to be common in young women of developed countries. Little is known however about body image preoccupation or the prevalence of dieting for weight control purposes in elderly women. The few available reports suggest that preoccupation with weight remain high in elderly women and that pressure to be thin drives normal weight older women to recurrent dieting. After a reminder of the nutritional vulnerability of the elderly recommendations to the health care professionals and health authorities are provided. An increased awareness of eating habits and weight preoccupation in elderly women is needed, since attitudes towards weight may influence the effectiveness of medical advice and health promotion campaigns. PMID- 10840474 TI - Interleukin-2, its receptor and nutrition in older adults: a review. AB - It is well recognized that malnutrition can impair immune function. Conversely, immune activity may influence measures of malnutrition, such as serum albumin and prealbumin. Interleukin-2 (lL-2) and its receptor are key components of immune function. Recent evidence has expanded our understanding of the interaction between nutrition and this cytokine system. particularly in older adults. (A cytokine is a protein that acts as a "hormone" regulator of the immune system.) This paper will summarize more recent findings regarding the relationship between nutrition and the IL-2/IL-2 receptor system. PMID- 10840475 TI - Enteral and parenteral nutrition in the seriously ill, hospitalized patient: a critical review of the evidence. AB - Providing nutritional support has become a standard component of managing seriously ill patients. While many studies have documented that providing nutrition support can change nutritional outcomes (amino acid profile, weight gain, nitrogen balance, etc.), there are limited da ta that demonstrate that nutrition support actually influences clinicall y important endpoints. The purpose of this paper is to systematically r eview and critically appraise the literature to examine the relationship between nutritional support and morbidity and mortality in the seriously ill patient. As a product of this critical appraisal of the evidence, clinical recommendations based on rules of evidence are put forward. PMID- 10840476 TI - Management of malnutrition in the elderly and the appropriate use of commercially manufactured oral nutritional supplements. AB - The risk of protein calorie malnutrition among the community dwelling elderly is high. If undetected or untreated malnutrition can raise risks of morbidity and mortality. Therefore, improving screening, assessment and treatment of malnutrition, particularly in ambulatory settings that serve the elderly is imperative. The aim of this paper is to briefly detail the diagnostic criteria for protein calorie malnutrition and discuss the special considerations necessary in evaluating risk and causes of malnutrition in the elderly. We conclude with a review of current management options and discuss the cost effectiveness and potential inappropriate use of commercially manufactured oral nutritional supplements. PMID- 10840477 TI - Clinical aspects of long-term enteral nutrition via percutaneous endoscopic gastrostomy (PEG). AB - Adequate enteral nutrition is a major medical goal in patients with inadequate oral food intake caused by various underlying diseases of benign and malignant origin. Due to its safe and technically simple route of placement the PEG has become the preferred enteral route of choice for enteral long-term nutrition world-wide. The incidence of PEG placement is still rapidly increasing. Based on the results of recently published large clinical studies the technical success rate is >99 % with a procedure-related lethal rate of about 0 %. Prospective clinical studies clearly prove the excellent individual subjective acceptance of this method by the patients. Indications, contra-indications, complications and technical procedures of PEG placement are described in detail in the present review. In general, long-term enteral feeding via PEG is accepted as a safe, effective, easy to practice and highly acceptable method with excellent long-term results and distinct improvement of nutritional status. Individual decisions for PEG placement should be considered much earlier and more frequently in appropriate patients to save and further improve quality of life in patients with inadequate oral food intake caused by various benign and malignant diseases. PMID- 10840478 TI - Dietary supplement use at the population level: recent experience from the 1994-5 British National Diet and Nutrition Survey: people aged 65 years and over. AB - The purpose of this article is to alert readers of this issue to the ongoing National Diet and Nutrition Survey series in mainland Britain, and to draw attention to the existence of information on the use of dietary supplements by different age-groups in the UK, and to some specific issues which need to be addressed, in order to achieve reliable estimates of supplement use from surveys of this type. PMID- 10840480 TI - Gastrostomy feeding in the elderly after acute dysphasgic stroke. PMID- 10840479 TI - Nutrition in the elderly hospital patient Nottingham studies. AB - Forty percent of elderly hospital admissions in the UK are undernourished, half severely so. Most continue to lose weight in hospital. not only because of disease, but also because of failure to identify and treat malnutrition and due to shortcomings in hospital food provision, upon which most patients depend for their nutritional care. Our studies have shown that more than 40% of food set before patients is left, and therefore wasted. This means that elderly patients are taking less than 70% of their energy (30-35 kcal/kg/day), and protein (1 dram/kg/day) requirements. Catering strategies, such as provision of smaller volume, high energy and protein density meals with snacks and, if necessary, proprietary oral supplements, have been shown not only to improve nutritional status of patients, but to result in improved clinical outcome. Our work has shown a relationship between malnutrition and loss of thermoregulation, which is reversed by appropriate feeding. We have also described the beneficial effects of overnight nasogastric tube feeding in undernourished patients with fractured femur. Like others, we have used a percutaneous endoscopic gastrostomy in the management of elderly patients with cerebrovascular and motor neurone disease, and have published audits of outcome in this field. PMID- 10840482 TI - Open invitation from the International Poverty and Health Network to all healthcare professionals. PMID- 10840481 TI - Signet ring cell carcinoma of a pulled-through sigmoid colon mimicking a primary invasive bladder tumor: case report and review of the literature. AB - Primary signet ring cell carcinoma of the urinary bladder and colon are rare disease entities that are aggressive, difficult to manage, and portend a poor prognosis. We present a case report of a 25-year-old man born with an imperforate anus who developed signet ring cell carcinoma of the pulled-through sigmoid colon that mimicked a primary invasive bladder tumor. Despite radical surgery and adjuvant radiation, the patient died of his disease 7 months after surgery. PMID- 10840483 TI - Reducing the burden of injury. PMID- 10840484 TI - Safety first: reducing exposure to allogeneic blood. PMID- 10840486 TI - A new medical school for regional Australia. PMID- 10840485 TI - Mortality from cardiovascular disease is too high outside capital cities. PMID- 10840488 TI - Excess coronary mortality among Australian men and women living outside the capital city statistical divisions. AB - OBJECTIVES: To compare rates of mortality from coronary heart disease (CHD) between populations living within and outside Australian capital city statistical divisions. DESIGN AND SETTING: Descriptive epidemiological study based on data for all residents of Australia aged 30-69 years who died between 1986 and 1996 in all States and Territories of Australia. MAIN OUTCOME MEASURES: Standardised mortality rates from all causes and coronary heart disease as coded by the Australian Bureau of Statistics, and estimated excess deaths in populations living outside capital city statistical divisions. RESULTS: Between 1986 and 1996, mortality from CHD declined by 46% in men and 51% in women, and accounted for 61% of the decline in mortality from all causes in men and 48% in women. More deaths than expected from acute myocardial infarction resulted in mortality rates from CHD up to 30% higher in men and 21% higher in women living outside the capital city statistical divisions, and accounted for an overall estimated excess of 3835 deaths from CHD in men (32% of excess deaths from all causes), and 1385 deaths from CHD in women (27% of excess deaths from all causes) over the 11-year study period. CONCLUSIONS: Although there were impressive declines in coronary mortality in all Australian States and Territories from 1986 to 1996, populations living outside capital cities continue to have higher death rates from CHD. These differences in mortality rates indicate a need for further research into factors which may influence mortality rates for CHD in rural and remote areas, and immediate measures to ensure optimal treatment of coronary risk factors and acute coronary events in such populations. PMID- 10840489 TI - Who's overweight? Comparison of the medical definition and community views. AB - OBJECTIVE: To investigate the extent to which people who are medically defined as overweight perceive themselves to be overweight. DESIGN: Secondary data analysis of the National Health Survey and the National Nutrition Survey conducted by the Australian Bureau of Statistics in 1995. PARTICIPANTS: 10,652 people aged 18 years and over (5076 men, 5576 women) in a multistage cluster sample of households throughout Australia. MAIN OUTCOME MEASURES: Body mass index (BMI) based on measured height and weight; self-reported perception of body weight (underweight, acceptable weight, or overweight). RESULTS: Among people with a measured BMI > or = 25, 49.3% of men (95% CI, 48.1%-50.5%) and 72.0% of women (95% CI, 70.8%-73.1%) considered themselves overweight. Among those with a measured BMI < 25, 3.4% of men (95% CI, 2.8%-4.1%) and 12.4% of women (95% CI, 11.4%-13.3%) considered themselves overweight. Older women were less likely to perceive themselves as overweight than younger women. The lowest BMI at which at least half the respondents considered themselves overweight was 26 to < 27 for women aged 18-59 years, and 28 to < 29 for older women and men. CONCLUSION: For many people, particularly men and older women, the meaning of "overweight" differs from the medical definition. Clinical and public health weight reduction programs which do not take this into account are unlikely to be successful. PMID- 10840487 TI - Use of interventions to minimise perioperative allogeneic blood transfusion in Australia. A survey by the International Study of Perioperative Transfusion (ISPOT) Study Group. AB - OBJECTIVE: To investigate use of interventions to minimise need for perioperative transfusion of allogeneic blood in surgical units in Australia. DESIGN: Two questionnaire-based surveys of practice. SETTING: All hospitals in Australia, 1996-1997. PARTICIPANTS: Survey 1: all Australian hospitals that have at least 50 beds and undertake surgery; Survey 2: surgical units identified as using the interventions. MAIN OUTCOME MEASURES: Reported rates of use of the various interventions (preoperative autologous donation, acute normovolaemic haemodilution [ANH], cell salvage, and drugs); use of guidelines; and perceptions about the appropriateness of current levels of use. RESULTS: Survey 1 was returned by 349 of 400 hospitals (87%) and Survey 2 by 324 of 578 surgical units (56%). Preoperative autologous donation was most widely used (70% of hospitals), most commonly in units performing orthopaedic or vascular surgery (65% and 37%, respectively). Cell salvage and ANH were used by 27% and 24% of hospitals, respectively, most often in units performing cardiothoracic (40% and 44%, respectively) and vascular surgery (29% and 15%, respectively). These three interventions were used significantly more in private than in public hospitals (P < 0.05). Use of printed guidelines was uncommon. Respondents considered that autologous transfusion techniques should be used more widely because of their perceived efficacy and concerns about safety of allogeneic blood. Perceived barriers to greater use included lack of surgeon or physician interest, uncertain scheduling of surgery in public hospitals and cost (cell salvage). Drugs to minimise blood loss were used by fewer than 10% of hospitals. CONCLUSIONS: Interventions to minimise the need for perioperative allogeneic blood transfusion (apart from drugs) are widely used in Australia. However, enthusiasm for intraoperative techniques of re-infusing autologous blood needs to be assessed against the evidence of their efficacy and cost-effectiveness. PMID- 10840490 TI - Social determinants of health: from observation to policy. AB - Differences in the health of individuals reflect inherent features of the societies in which they live. The way a society organizes itself and delivers wellbeing to its members are major determinants of health. Social, economic and political factors have an important influence on health and longevity. Social position and lifestyle only partially explain ill health. Psychosocial factors, such as a sense of isolation, deprivation or loss of control, are also important. Governments can reduce health inequalities by ensuring that all policies are assessed for their potential impact on the health of all sectors of society. PMID- 10840491 TI - Automatic external defibrillators: changing the way we manage ventricular fibrillation. AB - OBJECTIVE: To discuss recent developments in automatic defibrillation and to review the evidence that first-responders equipped with automatic external defibrillators (AEDs) improve survival from out-of-hospital cardiac arrest. DATA SOURCES: MEDLINE search from 1966 to 1999 (articles in English only) and examination of bibliographies. STUDY SELECTION: Published studies of out-of hospital cardiac arrest and first-responders equipped with AEDs. Studies had to have a control group and to report survival to hospital discharge from ventricular fibrillation (VF). DATA EXTRACTION: Six studies met the selection criteria (two prospective randomised trials, two prospective controlled trials, and one cohort study and one retrospective study, both with historical controls). DATA SYNTHESIS: A random effects meta-analysis of odds ratios for survival from VF. CONCLUSIONS: Meta-analysis suggests that equipping first-responders with AEDs increases the probability of survival to hospital discharge after out-of-hospital cardiac arrest (odds ratio, 1.74; 95% CI, 1.27-2.38; P < 0.001). However, most of the studies lacked sufficient power to draw definitive conclusions. Until the impact of wide deployment of AEDs is fully understood, first-responder defibrillation in Australia should only occur as part of coordinated multicentre research studies. PMID- 10840492 TI - Perspectives from a surgeon turned hospital chaplain. PMID- 10840493 TI - Neurological disorders. PMID- 10840494 TI - Transient ischaemic attacks and stroke. AB - Stroke is the third most common cause of death and a major cause of disability in Australia. Effective prevention is the most powerful strategy for reducing the burden of stroke. Major modifiable causal risk factors for stroke include hypertension, cigarette smoking, diabetes, atrial fibrillation, and carotid stenosis. Atrial fibrillation, in particular, is under-treated in the community; almost all patients should be prescribed warfarin or aspirin, depending on their absolute risk of stroke and risk of bleeding complications. Patients with suspected acute stroke should be referred immediately to a specialist stroke unit for urgent assessment and care by an interested, organised, multidisciplinary team of stroke experts. They should undergo immediate computed tomography brain scan and, if intracranial haemorrhage is excluded, be given aspirin (160-300 mg). Rehabilitation and secondary prevention of recurrent stroke should begin on day one after stroke. PMID- 10840495 TI - Risk management: how doctors, hospitals and MDOs can limit the costs of malpractice litigation. PMID- 10840496 TI - Risk management: how doctors, hospitals and MDOs can limit the costs of malpractice litigation. PMID- 10840497 TI - Risk management: how doctors, hospitals and MDOs can limit the costs of malpractice litigation. PMID- 10840498 TI - "Ute surfing": a novel cause of severe head injury. PMID- 10840499 TI - Patient attitudes to commonly promoted medical interventions. PMID- 10840500 TI - Vitamin D insufficiency and hyperparathyroidism in Perth blood donors. PMID- 10840501 TI - Universal periconceptional folate supplementation: chasing a dream? PMID- 10840502 TI - Australian adolescents in custody. PMID- 10840503 TI - Multilingualism and dyslexia: challenges for research and practice. AB - Over the last two decades there has been an expansion of activity and substantial progress in research on dyslexia and research on bilingualism and multilingualism. But the study of dyslexia has generally focused on monolingual learners and the study of bilingualism has tended to focus on speakers who do not have special educational needs. This paper will review the strands of research to date that have a bearing on multilingualism and dyslexia and attempt to identify the major challenges that face researchers and teachers. A satisfactory response cannot be developed without a full understanding of the impact that dyslexia has on language learning and the impact that multilingualism has on literacy learning. PMID- 10840504 TI - Issues in the assessment of reading disabilities in L2 children--beliefs and research evidence. AB - In bilingual and multilingual settings one is constantly challenged by the difficulty of teasing apart phenomena associated with normal second language (L2) reading acquisition from authentic warning signs of reading failure. The bulk of this paper focuses on a critical discussion of a cluster of beliefs that pertain to the issues concerning the diagnosis of reading disability in multilingual and bilingual settings among school children. Findings from available research on reading acquisition among bilingual children and research focusing specifically on the assessment of English-as-a-second language (ESL) children who might be at risk for reading disability are used to evaluate the validity of these beliefs. While some beliefs are supported by research, others are not. In particular, the research suggests that reliable diagnosis of dyslexia among ESL children can be achieved by examining within-language differences on various indices of basic reading skills such as phonological processing, and by noting a significant gap between oral and reading comprehension. PMID- 10840505 TI - An audit of the processes involved in identifying and assessing bilingual learners suspected of being dyslexic: a Scottish study. AB - The Commission for Racial Equality (Special Educational Needs Assessment in Strathclyde: Report of a Formal Investigation, CRE, London, 1996) highlighted the significant under-representation of bilingual children among pupils assessed as having specific learning difficulties/dyslexia. In this present study an audit was undertaken in order to explore issues arising from the Commission's report, initially using 53 schools from one education authority. This revealed an extremely low incidence of suspected dyslexia among bilingual pupils. A second study was carried out in a further nine education authorities, surveying 91 schools with bilingual pupils. The incidence of suspected dyslexia in bilingual pupils was found to be extremely low. Twenty-seven cases were examined. Most cases concerned pupils aged 7:0-9:0. Difficulties associated with conventional indicators of dyslexia are discussed. A wide variety of assessment approaches were reported and the use of first language (L1) assessment varied. The process of assessment tended to be lengthy and inconclusive. However, this report suggests that caution is necessary when considering dyslexia in the early stages of second language (L2) development. PMID- 10840506 TI - Dyslexia screening measures and bilingualism. AB - A series of measures used in a number of dyslexia screening tests was administered to groups of 7-8-year old English monolinguals and Sylheti/English bilinguals. Within these groups a subgroup of children was distinguished by poor spelling and reading in the absence of general ability, sensory, emotional or behavioural problems, i.e. specific literacy difficulties (SpLD). General ability (assessed by Raven's matrices), chronological age, male/female ratio and mono/bilingualism were controlled between SpLD and control groups. Screening measures assessed phonological skills, rapid naming, the ability to recite or repeat sequences of verbal and non-verbal stimuli, and visual and motor skills. Sample sizes were small owing to the selection criteria used and the small number of bilingual SpLD children identified. However, the results were encouraging in differentiating SpLD bilinguals from their peers, with the phonological measures in particular presenting consistent findings across bilingual and monolingual groupings. Those differences found between bilinguals and monolinguals are discussed in terms of a bilingual influence on the skills assessed or the reduced reliability of the measure. PMID- 10840507 TI - Systematic identification and intervention for reading difficulty: case studies of children with EAL. AB - Literacy underpins education. There is now very widespread concern over standards of literacy for children from multi-cultural backgrounds, who are learning English as a second or subsequent language, and who may have special educational needs. Research evidence suggests that the earlier children's difficulties can be identified, the more effective (and cost-effective) intervention will be, provided that the intervention is tailored to the child's abilities and skills. Nicolson and Fawcett have developed systematic procedures for identifying children at risk for reading difficulty, together with systematic teaching strategies to overcome reading difficulty. In this paper we present case studies of children with EAL (English as an additional language) drawn from a controlled study using computer interventions with secondary school children. Our findings indicate that children with EAL may be more resistant to remediation than some children with learning difficulties. The prognosis is more problematic for children with both EAL and dyslexia. PMID- 10840508 TI - Dynamic assessment and instructional strategies for learners who struggle to learn a foreign language. AB - In this paper the authors discuss how the concept of dynamic (cognitive) assessment and instruction might relate to the assessment and instruction of at risk foreign/second language learners. They describe its relevance to a diagnostic/prescriptive approach to instruction for teaching a foreign language to students with identified dyslexia and other at-risk students. They explain how to assess learners' knowledge of the native/foreign/second language through questions and guided discovery. Examples in German and English illustrate its application to foreign/second language instruction. PMID- 10840509 TI - Reflections on foreign language study for students with language learning problems: research, issues and challenges. AB - The study of foreign language (FL) learning for individuals who have found learning to read and write in their first language extremely problematic has been an under-researched area throughout the world. Since the 1980s, Leonore Ganschow and Richard Sparks have conducted pioneering research into the nature of difficulties, why they are encountered and how they can be minimized. In this paper the authors trace the development of their research on foreign language difficulties for students with language learning problems. They provide a summary of their findings and suggest new questions and directions for the field. PMID- 10840510 TI - College students with dyslexia: persistent linguistic deficits and foreign language learning. AB - The first of these two studies compared college students with dyslexia enrolled in modified Latin and Spanish classes and non-dyslexic students enrolled in regular foreign language classes on measures of foreign language aptitude, word decoding, spelling, phonological awareness and word repetition. The groups did not differ on age or grade point average. Analyses indicated that students with dyslexia performed significantly poorer on the foreign language aptitude measures as well as on both phonological tasks, reading and spelling. In the second study, students with learning disabilities who were enrolled in a modified Latin class were not significantly different from their peers in a regular Latin class on grade point average or on performance on a proficiency examination at the end of the second semester. The data suggest that while phonological processing deficits persist into adulthood, students with dyslexia are able to acquire appropriate skills and information to successfully complete the University's foreign language requirement in classes modified to meet their needs. PMID- 10840511 TI - Dyslexia and the learning of a foreign language in school: where are we going? AB - The difficulties which many dyslexic students encounter in the learning of the English language often extend to the learning of a foreign language in school. Although this problem has been acknowledged for some time, and although the learning of a modern foreign language is a core element in the Scottish curriculum, there has been little research into how modern languages can be presented to offer the best learning opportunities to dyslexic students. Dyslexic students are likely to benefit from a multisensory approach to the learning of a modern foreign language, and it seems likely that they will need to utilize similar strategies to those used for learning their first language. Strategies are discussed with a view to making modern language learning more appropriate for students with difficulties in learning. PMID- 10840512 TI - Teaching a foreign language using multisensory structured language techniques to at-risk learners: a review. AB - An overview of multisensory structured language (MSL) techniques used to teach a foreign language to at-risk students is outlined. Research supporting the use of MSL techniques is reviewed. Specific activities using the MSL approach to teach the phonology/orthography, grammar and vocabulary of the foreign language as well as reading and communicative activities in the foreign language are presented. PMID- 10840513 TI - Phonological representations, reading development and dyslexia: towards a cross linguistic theoretical framework. AB - This paper attempts to integrate recent research findings in phonological development, reading development and dyslexia into a coherent theoretical framework that can provide a developmental account of reading and reading difficulties across languages. It is proposed that the factors governing phonological development across languages are similar, but that important differences in the speed and level of phonological development are found following the acquisition of alphabetic literacy. The causal framework offered is at the level of a cognitive model, which may prove useful in organizing future cross-linguistic developmental work. PMID- 10840514 TI - Is English a dyslexic language? AB - McGuinness has suggested that there 'is no diagnosis and no evidence for any special type of reading disorder like dyslexia', and that poor teaching accounts for low levels of English literacy performance, rather than inherent personal deficits. Implicit in this is the assumption that some languages have simple grapheme-phoneme codes in which there is a one-to-one mapping, making them easy to teach and learn, while others have more complicated structures and are more difficult for teachers and students. There is now an increasing number of studies which demonstrate that readers in more transparent orthographies such as Italian, Spanish, Turkish, Greek and German have little difficulty in decoding written words, while English children have many more problems. Increasingly, lack of orthographic transparency in English is seen as having a powerful negative effect on the development of reading skills in English-speaking children. There is evidence that English-speaking children who fail to acquire reading skills may fall into two distinct categories: those who would succeed in languages, other than English, that have greater orthographic consistency; and those who would still have problems even with perfect orthographic transparency. The first, larger, group is let down by the interaction of poor teaching methods and an incomprehensible system of orthography. The present study examines word factors associated with poor spelling and reading that have been identified. Three factors account for the relative ease with which pupils can spell words: frequency of the word in the English language; length of the word; and the presence of 'tricky' letters or letter combinations. Data are presented illustrating the predictive model of spelling and reading which enables word difficulty to be calculated from the characteristics of English words. The implications the model has for teaching and learning English are elaborated, with reference to the possible benefits to be derived from mother-tongue teaching in British schools. PMID- 10840515 TI - Troglitazone withdrawn from market. PMID- 10840516 TI - Zonisamide approved for partial seizures. PMID- 10840517 TI - Cisapride to be taken off market, placed in controlled-access plan. PMID- 10840518 TI - July implementation set for hospital outpatient PPS. PMID- 10840519 TI - FDA commissioner notes actions against Internet sites selling prescription drugs. PMID- 10840520 TI - Effects of DTC ads on care examined, policies suggested. PMID- 10840521 TI - Glucosamine, chondroitin studies may overstate benefits in osteoarthritis, say authors. PMID- 10840522 TI - Supreme court strikes down FDA tobacco regulation. PMID- 10840523 TI - Managed care undervalues beneficial nonprescription drugs. PMID- 10840524 TI - Effect of a highly restrictive formulary on the average prescription cost at a community health center. PMID- 10840525 TI - Ethics and managed care formularies. PMID- 10840526 TI - Fortresses and formularies: drugs, ethics, and managed care. PMID- 10840529 TI - Therapeutic interchange in community pharmacies in Virginia. PMID- 10840527 TI - Risperidone for control of agitation in dementia patients. AB - Currently available research on the use of risperidone to manage agitation in patients with dementia is discussed. Dementia affects up to 70% of nursing-home patients, and more than 90% of them exhibit aggressive or agitated behavior or severe depression. Agitation includes combativeness, hyperactivity, disinhibition, wandering, and restlessness. Environmental interventions are preferred for mild symptoms; medications are the treatment of choice for severe manifestations. Traditional neuroleptics have been the mainstay of treatment for agitated behavior in persons with dementia, but these agents have limited efficacy and are associated with high rates of adverse effects, including worsening of already poor cognitive functioning. Although the literature on the use of risperidone in elderly patients with dementia consists largely of uncontrolled trials, case reports, and chart reviews, it appears that this agent is effective for managing agitation in this population and does so with a low frequency of extrapyramidal symptoms (EPS). Risperidone may also be useful for treating acute agitation in patients with a high risk of EPS and for long-term treatment of "sundowning" (agitation and confusion starting in the late afternoon and worsening at night). A low initial dosage that is gradually adjusted upward is recommended. Risperidone appears effective in controlling agitation in patients with dementia and has a relatively benign adverse-effect profile, but more clinical trials are needed to elucidate its role for this indication. PMID- 10840528 TI - Antimicrobial treatment patterns, resource utilization, and charges associated with acute sinusitis in asthma patients. AB - Antimicrobial treatment patterns, resource utilization, and charges associated with acute sinusitis in patients with asthma were studied. Asthma patients with at least one claim for acute sinusitis were identified by International Classification of Diseases, 9th Revision code in data for 1992-1994 from a large health care insurer in New England. A sinusitis episode was defined on the basis of the pattern of antimicrobial prescribing. Antimicrobials used to treat sinusitis, rates of resource utilization, and overall charges for therapy were determined. A total of 2,633 sinusitis episodes were identified in the records of 34,348 asthma patients. The most frequently prescribed antimicrobial for initial treatment of acute sinusitis was amoxicillin trihydrate (32.2% of patients). Initial therapy was successful in 2,199 episodes (83.5%), initial therapy failed in 250 episodes (9.5%), and relapse occurred in 184 episodes (7.0%). Successfully treated patients had fewer outpatient visits on average (1.42 per episode) and fewer laboratory and diagnostic tests (0.046) than did patients in whom initial therapy failed (1.76 and 0.056 per episode, respectively) or patients who relapsed (1.68 and 0.049). The mean total charge for sinusitis care was $147.61 per episode for successfully treated patients, $242.95 per episode for patients unresponsive to treatment, and $205.49 per episode for patients who relapsed. Antimicrobial treatment of acute sinusitis varied widely in patients with asthma. Resource utilization was associated with the success of treatment. PMID- 10840530 TI - Clinical trials in pharmacogenetics and pharmacogenomics: methods and applications. AB - Clinical and other aspects of pharmacogenetics and pharmacogenomics are discussed. Pharmacogenetics is the study of the impact of heritable traits on pharmacology and toxicology. An extension of pharmacogenetics is the discovery that genetic polymorphisms have the potential to affect a drug's action. The interplay of genotype and drug efficacy has been defined as pharmacogenomics. For most drugs, variations in patient response have until recently been considered a result of pharmacokinetic rather than pharmacodynamic differences. However, it now seems that pharmacodynamic variability in humans is large, reproducible, and usually more pronounced than pharmacokinetic variability. Some examples of the impact of pharmacogenomics on pharmacokinetics involve cytochrome P-450 isoenzymes, dihydropyrimidine dehydrogenase, and thiopurine methyltransferase; some examples of the impact on pharmacodynamics involve cholesteryl ester transfer protein, angiotensin-converting enzyme, and serotonin transporter. There are no specific statistical techniques for analyzing data from pharmacogenomic clinical trials. However, a tabulated relationship for the determination of the maximum possible gain in response rate for the highest-responding genotypic subgroup of patients is provided as an aid to determining whether it is worth having a pharmacogenomic strategy for a given drug. Ethical issues in pharmacogenomics tend to be based on the general concern that the ability to diagnose a genetic disorder before any treatment is available does more harm than good to the patient. Pharmacogenomic approaches to drug discovery and delivery have been recognized by FDA. Pharmacogenomics cannot improve the efficacy of a given drug, but it helps in selecting patients who are likely to respond well. Pharmacogenomics provides a view of drug behavior and sensitivity useful to improving the efficacy of drug development and utilization. PMID- 10840531 TI - Comparing medication use in two hemodialysis units against national dialysis databases. PMID- 10840532 TI - An intranet-based formulary management system. PMID- 10840533 TI - [Trauma patients--establishing comparable data sets]. PMID- 10840534 TI - [Recommendations for uniform documentation according to trauma severity--the Utstein style. An international initiative of the International Trauma Anesthesia and Critical Care Society (ITACCS)]. PMID- 10840535 TI - [Use of the sitting position in neurosurgery. Results of a 1998 survey in Germany]. AB - OBJECTIVES: This 1998 survey was carried out on the use of the sitting position for neurosurgical procedures in the posterior fossa and operations of the craniospinal and cervical spine region by the dorsal approach. In addition, anesthetic management of the sitting position and the compliance with recommendations of the Neuroanesthesia Study Group of the German Society of Anesthesiology and Intensive Care Medicine (DGAI) published in 1995 were investigated and compared to results of a 1995 survey. METHODS: A questionnaire was sent to 152 departments of anesthesiology in Germany providing anesthesia for neurosurgical procedures. 85 institutions (56%) responded to the survey, data from 78 hospitals were enrolled into the study. The sitting position was preferred for posterior fossa surgery by 45% of the neurosurgeons, for craniospinal operations by 35% and for cervical spine surgery by the dorsal approach by 39%. To 97% of the institutions the recommendations of the Neuroanesthesia Study Group of the DGAI were well known, 19% modified their anesthetic approach due to these recommendations. Recommendations of the Study Group on neuro-monitoring, in particular on the use of ultrasound (precordially or transoesophageally) for the detection of venous air embolism were followed by all institutions. 45% of the participants of the study preoperatively undertook diagnostic measures to preclude a probe-patent foramen ovale which predisposes the patient to paradoxical air embolism. CONCLUSIONS: The survey demonstrates that the use of the sitting position in German neurosurgery is still high when compared to other Western countries, but a tendency for decline over last 3 years can be observed from our data. In addition, our data appears to indicate a positive effect of the Study Group's recommendations on anesthetic management of the sitting position in neurosurgery. PMID- 10840536 TI - [Tropisetron for prevention of nausea and vomiting in children undergoing tonsillectomy and/or adenoidectomy]. AB - BACKGROUND: Postoperative nausea and vomiting (PONV) after tonsillectomy is a common problem in children. Tropisetron is a new 5HT3 receptor antagonist and is successfully used in paediatric patients receiving cancer therapy. The aim of the study was to assess efficacy and safety of a single intravenous dose of tropisetron for prevention of PONV in paediatric patients at risk for postoperative vomiting. METHODS: In a randomised, double-blind, placebo controlled trial, we studied 98 children aged 2-12 years undergoing tonsillectomy or adenotonsillectomy. Patients received placebo or tropisetron 0.1 mg (= 0.1 ml)/kg body weight immediately after induction of anesthesia. A standard general anesthetic technique (Sevoflurane/N2O/O2 without neuromuscular blockers or opioids) was used. Perioperative vital signs, grade of sedation and episodes of postoperative nausea and vomiting were recorded. RESULTS: No vomiting episodes occurred in 65.3% of the tropisetron treated patients compared to 34.7% of the placebo group (p = 0.0024). Only 10.2% of the tropisetron treated patients vomited more than 3 times compared to 22.4% of the control patients (p = 0.0004). The need for antiemetic rescue medication was significantly lower in the study group (10.4%) compared to 28.6% (p = 0.025). No significant adverse effects of the study medication were shown. CONCLUSION: A single intravenous prophylactic dose of tropisetron effectively reduces the incidence of PONV during the first 24 postoperative hours after tonsillectomy and/or adenoidectomy. Because of the low incidence of adverse effects, the prophylactic use of tropisetron seems to be safe and justified in paediatric surgical patients at high risk for postoperative vomiting. PMID- 10840539 TI - [From isoflurane to perfluorohexane? Perfluorocarbons--therapeutic strategies in acute lung failure]. AB - The introduction of Perfluorochemicals into medicine and especially into the treatment of severe lung injury is a fascinating scientific task. Many recall the famous experiments from Clark et al. in 1966 when he demonstrated "liquidventilation with perfluorocarbons" in the mammal species for the first time. After this hallmark, perfluorocarbons were subsequently introduced in research of acute lung injury by the techniques of Total- and Partial-Liquid Ventilation (TLV; PLV). Perfluorocarbons (saturated organofluorids) have unique chemical and physical properties which made them attractive substances for intraalveolar application. The strong C-F bindings in the perfluorocarbon molecules are responsible for their chemical stability, biochemical inertness, high capacity to dissolve respiratory gases, low surface tension and high vapor pressures. Furthermore, the high density of the PFC lead to radio-opacity and their distribution to dependent lung areas. The efficacy of PFC liquid, applied by TLV/PLV has been demonstrated in numerous animal studies using different models of acute lung injury. Currently, several mechanisms of action of perfluorocarbon fluids in acute lung injury are discussed: recruitment of atelectatic alveoli, prevention of endexpiratory collapse of alveoli ("liquid PEEP"), redistribution of perfusion, oxygen transport, surfactant like effects and decrease of inflammation. Since total liquid ventilation has been used only in experimental models of lung injury, partial liquid ventilation has been introduced successfully into clinical trials (phase I-II). However, the results of the first randomised, controlled study of PLV in 90 adult patients suffering from severe respiratory failure (ALI/ARDS) showed no differences between PLV and conventional treatment. Furthermore, the instillation of relatively large amounts of liquid into the lungs poses several technical challenges and may be associated with complications such as liquithoraces, pneumothoraces and hypoxia. Since mammal lungs are evolutionary specialised to gas exchange using atmospheric oxygen, the application of liquids, even if they transport respiratory gases very well is not physiologic. To overcome these unwanted side effects, we developed a technique of perfluorocarbon vaporisation in analogy to the application of inhalation anaesthetic agents. After resolving some technical issues, this application technique was used successfully in an animal model of acute lung injury. Vaporisation of perfluorohexane in a concentration of 18 Vol.% of inspired gas improved significantly oxygenation and lung compliance. Though these results are promising, mechanisms of action, dose-efficacy relation, surfactant perfluorocarbon interaction or anti-inflammatory effects of vaporised perfluorohexane are still unclear. These questions need to be clarified before this technique can be applied clinically. However, the inhalation of vapor, a technique already familiar to anaesthesiologists should avoid risks of large amounts of fluids in the bronchoalveolar space. Furthermore, this technique can be administered by established anaesthetic equipment with the advantage of exact dosing, continuous monitoring, and demand application in a way near to clinical routine. PMID- 10840537 TI - [The effect of aprotinin and tranexamic acid on fibrinolysis and thrombin generation during cardiopulmonary bypass]. AB - OBJECTIVE: Antifibrinolytic drug therapy has proved to be effective in reducing blood loss associated with cardiac surgery and cardiopulmonary bypass (CPB). Concerns remain regarding the risk of enhancing thrombosis. In the present study we investigated the effect of aprotinin (AP) and tranexamic acid (TA) on fibrinolysis and thrombin generation during CPB. METHODS: 60 patients undergoing coronary artery bypass graft surgery were randomised in 3 groups. They received either aprotinin ("high-dose-scheme"), tranexamic acid (2 g/h) or no antifibrinolytic therapy (control group). Collection of blood was performed at 7 pre-, intra- and postoperatively predetermined intervals. Fibrinolytic activity was determined by measuring concentrations of D-dimer, thrombin generation by the measurement of thrombin-antithrombin III complex (TAT). RESULTS: There was no significant increase of D-dimers in the AP or TA group. D-dimer concentration in the control group increased significantly after starting CPB. Comparing with the control group, thrombin generation in the AP group was significant less, while TA group produced significantly higher values. CONCLUSION: After the administration of AP for cardiac surgery we observed reductions in both intraoperative fibrinolysis and thrombin generation. In case of TA suppression of fibrinolytic activity in the absence of concomitant reduction in thrombin generation occurred. These results suggest that TA could potentiate a hypercoagulable state with the risk of thrombosis in the perioperative setting. PMID- 10840538 TI - [Cognitive impairment in the early postoperative period after remifentanil propofol and sevoflurane-fentanyl anesthesia]. AB - OBJECTIVE: In ambulatory anaesthesia the time required to recover from cognitive impairment should be as short as possible. The aim of this study was to compare the early cognitive recovery after remifentanil/propofol (R/P) and sevoflurane/fentanyl (S/F) anaesthesia. METHODS: Sixty patients scheduled for elective gynaecological laparoscopy and 24 female volunteers tested for the assessment of learning effects were investigated. After praemedication with midazolam anaesthesia was induced with propofol, atracurium and either 1 microgram/kg fentanyl or 1 microgram/kg remifentanil. For maintenance 0.25 microgram/kg/min remifentanil and 0.6 mg/kg/min propofol (R/P) or 1.7 vol% sevoflurane (S/F) were given. Both groups were ventilated with 30% oxygen in air and received metamizol for postoperative analgesia. Verbal Learning Test, Stroop Colour and Word Interference Test, Digit Symbol Substitution Test and Four Boxes Test were performed the day before surgery and 30 min, 1 h, 2 h and 4 h after termination of anaesthesia. RESULTS: For remifentanil/propofol cognitive function was still impaired 2 h (Verbal Learning) and 4 h (Stroop, Digit Symbol Substitution and Four Boxes Test) after termination of anaesthesia. After sevoflurane/fentanyl anaesthesia cognitive impairment lasted the same duration in Four Boxes Test, but shorter in Stroop and Digit Symbol Substitution and could not be found in Verbal Learning Test. CONCLUSION: The duration of cognitive impairment in the early postoperative period differed by the test procedures and the anaesthetic procedures used in this investigation. Recovery appeared to be faster after sevoflurane/fentanyl than after remifentanil/propofol at least in aspects of cognitive function. PMID- 10840541 TI - [Induction of anesthesia for a patient with sleep apnea syndrome]. AB - Based on a case report, we offer brief guidelines on the perioperative management of patients with Sleep-Apnea-Syndrome (SAS) who present with a high incidence of a difficult airway and a high risk of respiratory depression during the perioperative period. A 39 year old male patient with a body mass index of 34.22 kg/m2 and receiving continuous-positive-airway-pressure-(CPAP) therapy for known SAS was scheduled for elective plastic surgery. After induction of anaesthesia and direct laryngoscopy no adequate airway could be established and the patient became hypoxic, hypercapnic and developed hypotension and bradycardia. With the use of a laryngeal mask airway the patient was stabilized and did not show neurologic sequale after immediate awakening. The following fiberoptic intubation of the awake patient, still showing tendency of upper airway obstruction, confirmed the difficult anatomical structures. The subsequent general anesthesia was uneventful. The patient received CPAP therapy and was monitored during the first postoperative night in the Intensive Care Unit. He made an uneventful recovery. He was advised to have regional anaesthesia or planned fiberoptic intubation, where possible, in the case of further anesthetic intervention. SAS has major implications for the anaesthesiologist and whenever patients exhibiting the high risk factors (obesity, male sex, history of intense snoring, impaired daytime performance, nonrefreshing daytime naps) are presented for surgery this condition should be considered. Elective surgery should be postponed until after adequate examination and treatment when necessary. Patients with SAS should always be suspected of having cardiopulmonary dysfunctions such as hypertension, cardiac dysrhythmia or cor pulmonale. It is most important to avoid sedative premedication, to initiate CPAP therapy preoperatively, to encourage regional anaesthesia if possible and to ensure close monitoring over the complete perioperative period. Planned fiberoptic intubation, preferably with surgical personnel available for an emergency airway, is a safe method for the induction of anaesthesia. Postoperatively, patients are at high risk from respiratory depression, even in the awake state. Postoperative opioid analgesia, no matter what route, should only be given under close monitoring. Independently of regional or general anaesthesia there is an increased risk of respiratory depression in the middle of the first postoperative week, suspected to be caused by the catching up on lost REM-sleep, due to shifts in the normal sleep pattern during the first postoperative days. PMID- 10840540 TI - [Severe diving accidents: physiopathology, symptoms, therapy]. AB - Decompression injuries are potentially life-threatening incidents, generated by a rapid decline in ambient pressure. Although typically seen in divers, they may be observed in compressed air workers and others exposed to hyperbaric environments. Decompression illness (DCI) results from liberation of gas bubbles in the blood and tissues. DCI may be classified as decompression sickness (DCS) or arterial gas embolism (AGE), depending on where the gas bubbles lodge. DCS occurs after longer exposures to a hyperbaric environment with correspondingly larger up-take of inert gas. DCS may be classified into type 1 with cutaneous symptoms and musculoskeletal pain only or type 2 with neurologic and/or pulmonary symptoms as well. AGE usually results from a pulmonary barotrauma, and with cerebral arterial involvement, the symptoms are similar to a stroke. The most important therapy, in the field, is oxygen resuscitation with the highest possible concentration and volume delivered. The definitive treatment is rapid recompression with hyperbaric oxygen therapy. Additional therapeutic measures are discussed. PMID- 10840542 TI - [Arterial air embolism as a complication to blunt thoracic trauma. Remarks on the paper of S. Gotz-Albrod et al. Anaesthesist (1999) 48:452-454]. PMID- 10840543 TI - [In Process Citation] PMID- 10840544 TI - [Clearing the epidural catheter in the delivery room]. PMID- 10840545 TI - [Head-brain injury and cerebral hypoxia. Diagnosis--monitoring--therapy]. AB - The main reason for posttraumatic secondary brain damage is cerebral hypoxia. Both, severity and duration of hypoxia are crucial in determining wether irreversible cerebral infarction will occur or not. For the clinical routine, the diagnosis of hypoxia is indirectly made by low CPP, low jugular-venous oxygen saturation (SjO2) or low tissue PO2. To minimize misleading false negative SjO2, the CT-Approach for the side of monitoring and calculation of arterial-jugular venous lactate content for detection of anaerobic metabolism is recommended. Targeted treatment of hypoxia according to the underlying cause is mandatory. Primary goal is to increase cellular oxygen delivery by correction of low arterial oxygen content and elevation of regional CBF. Within the autoregulatory range decreasing CPP causes vasodilation and increasing CPP vasoconstriction with increasing or decreasing cerebral blood volume respectively. Initially elevation of the lower autoregulatory threshold often requires CPP 70 mmHg. Targeted treatment of intracranial hypertension must avoid decreasing CPP. In the early posttraumatic phase prevention of cerebral hypoxia relies on management of CBF by means of CPP and cerebral vascular resistance. Thereafter targeted treatment of intracranial hypertension caused by cerebral edema and hypervolemia are increasingly important. PMID- 10840547 TI - [Treatment of anesthesia-induced atelactasis]. AB - During general anaesthesia even healthy lungs tend to collapse. Thus, up to 20% of previously functional lung tissue may be lost for gas exchange. It should be advantageous to treat this pathologic condition. After explaining the clinical problem of lung collapse, the concept of opening these lungs and keeping them open will be discussed. Some results of the first randomized clinical trials on intraoperative lung recruitment will be presented. Finally, a systematic description of all treatment steps tries to provide the anaesthesiologist with a useful practical guide for applying the "alveolar recruitment strategy" in their daily care of patients undergoing general anaesthesia. PMID- 10840546 TI - [Cerebral protection. The role of nutritional therapy]. AB - Prevention of secondary cerebral insults has the highest priority as far as therapeutic interventions of the patient with brain lesions are concerned. Patients with cerebral lesions have to overcome both, cerebral and systemic insults. The intensity of the neuronal injury determines the grade of hypermetabolism. Optimal metabolic and nutritional therapy for patients with cerebral lesions should be accomplished to minimize secondary brain damage. Frequently, the systemic hypermetabolic response is associated with cerebral ischemic metabolism. Therefore systemic blood glucose levels should be less than 150 mg/dl to prevent intracellular anaerobic accumulation of lactate. Individual utilization capacity of substrates is determined by the grade of hypermetabolism. Substrate load has to be adapted to the individual utilization capacity to avoid side effects of nutritional therapy like substrate and volume overload, imbalances of electrolytes as well as enhanced application of excitatoric substrates. In addition, whenever possible enteral nutrition should be applied to profit from reduced bacterial translocation and improved glucose hemostasis. Oxygen radical production and lipid peroxidation are important pathophysiologic mechanisms concerning cerebral lesions. More recent data show reduced antioxidative status in patients with brain injuries which favors lipid peroxidation. Further studies must be carried out to evaluate the potential neuro protective effect of an antioxidative nutritional regimen in patients with cerebral lesions. PMID- 10840548 TI - [Acute kidney failure. Physiopathology--clinical diagnosis--therapy]. AB - Acute renal failure (ARF) is characterized by an acute decrease in glomerular filtration rate (GFR). ARF complicates 4% to 23% of intensive care unit admissions, and is associated with a mortality of approximately 50% among critically ill patients. In the intensive care setting the term ARF is usually applied to acute tubular necrosis (ATN), a form of intrinsic ARF caused by ischemia or nephrotoxins. Pathophysiological mechanisms involved in the decline in GFR include tubular obstruction caused by detachment of tubular epithelial cells from the basement membrane and back-leak of glomerular filtrate as a consequence of disruption of the epithelial cell layer. Vascular mechanisms involved in the pathophysiology of ATN are vasoconstriction due to an imbalance between vasoconstrictive and vasodilatory mediators and vascular obstruction caused by cell aggregation. Currently, there is no real time method to monitor renal function comparable to the real time monitoring of blood pressure or arterial oxygen saturation. Urinary output does not reflect glomerular filtration which may be critically reduced despite normal urine volumes and creatinine clearance still provides the clinically most applicable estimate of GFR. Tubular function can be assessed using the fractional excretion of sodium or the ratio of urinary and serum osmolality; both parameters can be obtained from spot samples of urine and serum and no urinary sampling period is necessary. However, both parameters are strongly affected by the administration of loop diuretics and high fluid and sodium inputs which are common in the intensive care unit. We determined the day to day variability of creatinine clearance, fractional excretion of sodium and the urinary to serum osmolality ratio in critically ill patients without renal dysfunction (i.e. creatinine clearance in the normal range) and found differences of 16% for creatinine clearance, 79% for fractional excretion of sodium and 22% for urinary to serum osmolality ratio. Treatment of ARF is mainly supportive and there is no clinically accepted therapy that attenuates the course of ATN. Treatment of the underlying disease and renal replacement therapy are the main options for the treatment of patients with ARF. In critically ill patients continuous venovenous hemo(dia)filtration is the first choice because it provides more hemodynamic and metabolic stability than intermittent therapy. Acute life-threatening hyperkalemia is an indication for intermittent hemodialysis because of the higher efficacy of dialysis in the clearance of low molecular weight substances. PMID- 10840549 TI - [Strategies for the use of amino acids. Catabolism and retention]. AB - Extensive Catabolism is a hallmark of patients with acute renal failure (ARF) complicating critical illnesses. Catabolism is due to dysregulation of protein metabolism as well as a consequence of diminished renal excretion and renal replacement therapy (RRT). Inadequate nutritional support predisposes patients to malnutrition and increased mortality risk. Since the catabolic rate varies widely in ARF patients and can not be predicted by clinical parameters, direct quantification of the protein catabolic rate should be performed regularly. The urea nitrogen appearance rate (UNA) is a valid and reproducible estimate of nitrogen balance in critically ill patients undergoing continuous RRT. Amino acid losses up to 50 g/day and protein losses up to 15 g/day occur during continuous RRT and need to be compensated for. In order to achieve neutral or positive nitrogen balance a nutritional regimen providing 1.5-2.0 g protein/kg/day and 25 35 kcal/kg/day may be required. Since glutamine losses during continuous RRT may exceed 4 g/day, glutamine supplementation (0.3-0.5 g/kg/day) appears to be recommendable. PMID- 10840550 TI - [Burn trauma. Preclinical and clinical care from an anesthesiologist's point of view]. PMID- 10840551 TI - [Neuromuscular monitoring: toy or indispensible tool]. PMID- 10840552 TI - [Intraoperative monitoring: necessary, meaningful or superfluous]. PMID- 10840553 TI - [Standards of various national societies and their actual use in practice]. PMID- 10840554 TI - [The choice of test muscles]. PMID- 10840555 TI - [The choice of stimulation patterns]. PMID- 10840556 TI - [Neuromuscular monitoring following anesthesia induction]. PMID- 10840557 TI - [Intraoperative monitoring of relaxation]. PMID- 10840558 TI - Residual neuromuscular blockade. Incidence and relevance. AB - Residual neuromuscular block is a major risk factor behind critical events in the immediate postoperative period. Residual weakness due to muscle relaxants is seen in more than thirds of postoperative patients with ventilatory failure and hypoxia. Residual neuromuscular block should therefore be regarded as a serious adverse event in the same way as we regard ventilatory depression due to opioids and anaesthetic agents. This presentation aim to clarify our present knowledge and shortcomings in the field of residual neuromuscular blockade. PMID- 10840559 TI - [Monitoring of residual neuromuscular block]. PMID- 10840560 TI - [Objective monitoring in everyday clinical use]. PMID- 10840561 TI - [Neuromuscular monitoring in myasthenia gravis]. PMID- 10840562 TI - [Neuropathies: the effect of relaxography and neuromuscular function]. PMID- 10840563 TI - Radiation dose to patients from radiopharmaceuticals (addendum 2 to ICRP publication 53). AB - A joint Task Group of ICRP Committees 2 (Doses from Radiation Exposures) and 3 (Protection in Medicine; lead Committee for this report) has prepared a compilation of data on radiation dose to patients from radiopharmaceuticals. The report provides biokinetic models, absorbed doses, and effective doses, using ICRP Publication 60 dosimetry, for 10 new radiopharmaceuticals: [Methyl 11C]thymidine; [2-11C]thymidine; 14C urea (incl. carbon dioxide and bicarbonate); 15O water; 99mTc HIG, Pertechnegas, Technegas, and tetrofosmin; and 111In HIG and octreotide. It also provides recalculated dose data for the 19 most frequently used radiopharmaceuticals from ICRP Publication 53, using ICRP Publication 60 dosimetry, viz. 18F FDG; 51Cr EDTA; 67Ga citrate; 75Se SeHCAT; 99mTc DMSA, DTPA, RBC, IDA, large colloids, WBC, MAA, non-absorbable markers, pertechnetate, and phosphates and phosphonates; 123I Hippuran and MIBG; 131I Hippuran and NP59; and 201Tl thallous ion. Printing errors detected in ICRP Publication 53 are also listed. Furthermore, the report reproduces with minor corrections and updates, and therefore supersedes, the information on 6 radiopharmaceuticals given in Addendum 1 to ICRP Publication 53: 3H neutral fat and fatty acids; 14C neutral fat and fatty acids; 68Ga EDTA; and 99mTc HM-PAO, MAG3, and MIBI. There is an integrated index to all radiopharmaceuticals treated in ICRP publications so far, including a listing of effective doses per unit activity administered to adults. This issue of the Annals of the ICRP also includes an Addendum to ICRP Publication 72 concerning age-dependent doses to members of the public from intakes of radionuclides. PMID- 10840564 TI - Quarantine defences in Australia's north. PMID- 10840565 TI - Docking is painful. PMID- 10840566 TI - Generalised demodicosis in dogs, clinical perspective. PMID- 10840567 TI - Treatment of canine adult-onset demodicosis. PMID- 10840568 TI - Generalised demodicosis in the dog: the unresponsive or recurrent case. PMID- 10840569 TI - Zygomycotic proventriculitis and ventriculitis in ostriches (Struthio camelus) with impaction. AB - Three young ostriches (Struthio camelus), aged 4 months, were found to have zygomycotic proventriculitis and ventriculitis associated with impaction. Clinical signs were anorexia, chronic weight loss, weakness and lethargy followed by scant faeces for seven days. Proventriculi and ventriculi from birds were full of masses of hay, grass, leaves and other fibrous materials in combination with sand, gravel and plastic. Erosions and haemorrhagic ulcers of varying number and severity were present in the mucosae of both organs involved. Mucosal lesions were characterized by haemorrhagic necrosis. Throughout the affected mucosae there were 5 to 12 microns wide rarely-septated fungal hyphae with non-parallel walls, irregular branching and occasional globoid distentions, typical of Zygomycetes. Zygomycotic proventriculitis and ventriculitis secondary to impaction was diagnosed. PMID- 10840570 TI - Association between milk production, somatic cell count and bacterial dermatoses in three dairy cattle herds. AB - OBJECTIVE: To determine the correlations between three bacterial dermatoses in cattle, milk production and bulk-milk somatic cell count (BMSCC). DESIGN: Field observations in three dairy cattle herds. METHODS: Milk production, BMSCC, fertility and all herd diseases were recorded by computerised dairy management systems. Each herd was visited twice weekly and the clinical signs, course of diseases and morbidity and culling rates were noted. Bulk-tank milk was sampled twice monthly and analysed for somatic cell count. Bacteriological and histological examinations were carried out from samples collected from affected animals in the respective herds. RESULTS: The acute exudative form of dermatophilosis was diagnosed only in first-calving cows. The morbidity rate was 53% and the culling rate was 16% of the affected animals. The BMSCC increased by a factor of 2.4 times, and there was an average loss of milk production of 30%/cow/day in affected animals. Ulcerative lymphangitis was diagnosed in first calving cows (22%) and older cows (15%). The culling rate was 28%. The BMSCC increased by a factor of 17.3 times, and the average loss of milk production was 5.5%/affected animal/day. Papillomatous digital dermatitis (PDD) was diagnosed in first-calving cows (25%) and in older cows (18%). The culling rate was 8%. The BMSCC increased by a factor of two times, and the average loss of milk production was 1.7%/affected animal/day. CONCLUSIONS: The correlations between three skin diseases (ulcerative lymphangitis, dermatophilosis, papillomatous digital dermatitis), milk production and BMSCC have been found to be unfavourable. PMID- 10840572 TI - Cutaneous phycomycosis in two horses. PMID- 10840571 TI - Acute bronchopneumonia associated with Mycobacterium fortuitum infection in a dog. AB - Mycobacterium fortuitum was isolated in a sample of bronchial fluid collected by transtracheal aspiration from a 1-year-old Corgi dog with a productive cough of 10 days' duration and with radiographic and cytological features of acute suppurative bronchopneumonia. The dog responded favourably to intravenous gentamicin and cephalexin for three days and a six week course of oral ciprofloxacin. Saprophytic mycobacterial pneumonia should be considered in cases of severe pulmonary consolidation in young dogs. PMID- 10840573 TI - Neospora caninum infections in Australia and New Zealand. AB - OBJECTIVE: To review the current state of knowledge of Neospora caninum infections with particular reference to Australia and New Zealand. PROCEDURE: Several databases were searched electronically including Medline, Current Contents, Vet CD using several key words (Neospora caninum, neosporosis, abortion, cattle, dogs) and authors names. References in original articles were also traced and use made of the author's own original research in the field. CONCLUSION: N caninum is recognised worldwide and is also widespread, in particular in dairy cattle, in Australia and New Zealand. It has been reported in both countries retrospectively (in dogs) from the early 1970s. Abortion storms in dairy herds appear to be the most common feature. Recent reports indicate that the dog is the definitive host, in which the sexual cycle is completed. Further studies are however required to establish important aspects of the epidemiology, such as mode of transmission. In the absence of an effective vaccine, the most effective control strategy is selective culling of infected animals and the prevention of access of dogs to expelled placentas and foetuses and, possibly, to raw beef. PMID- 10840574 TI - Seroprevalence of Neospora caninum infection following an abortion outbreak in a dairy cattle herd. AB - OBJECTIVE: To investigate the seroprevalence of Neospora caninum infection in a commercial dairy cattle herd, 15 months after detection of an abortion outbreak. PROCEDURE: Sera from the whole herd (n = 266) were examined for N caninum antibodies by indirect fluorescent antibody test (IFAT) and immunoblot analysis. Herd records were reviewed to collate serological results with abortion history, proximity to calving, and pedigree data. RESULTS: The seroprevalence of N caninum infection was 24% (63/266) for IFAT titre > or = 160, 29% (78/266) for immunoblot positive (+ve), and 31% (82/266) for IFAT > or = 160 and/or immunoblot +ve; 94% (59/63) of animals with IFAT > or = 160 were immunoblot +ve. The association between seropositivity (IFAT > or = 160 and/or immunoblot +ve) and history of abortion was highly significant (P < 0.001); the seroprevalence was 86% (18/21) in aborting cows, compared with 30% (50/164) in non-aborting animals. The abortion rate for seropositive cows was 26% (18/68) compared with 3% (3/117) for seronegative animals. IFAT titres of infected cows were higher within 2 months of calving than at other times (P < 0.001). The association between seropositivity in dams and daughters was highly significant (P = 0.009). CONCLUSIONS: The abortions were associated with N caninum infection and there was evidence of reactivation of latent infection close to calving and congenital transmission of infection. Immunodominant antigens identified by immunoblots may prove useful for improved diagnostic tests. PMID- 10840575 TI - Use of multilocus enzyme electrophoresis to distinguish clinically important strains of Staphylococcus intermedius from the skin of dogs. AB - AIMS: To use multilocus enzyme electrophoresis to determine the genetic structure of Staphylococcus intermedius from normal skin of dogs and those isolated from a variety of disease conditions and to distinguish clinically important strains in dogs. METHODOLOGY: The diversity amongst 129 isolates of S intermedius from the skin and mucosa of 32 healthy dogs and 120 isolates from diseased sites in 120 individual dogs was examined using multilocus enzyme electrophoresis. Associations among ETs were examined to determine the diversity of isolates. RESULTS: Twenty two ETs were distinguished comprising 21 containing isolates from diseased sites and 11 containing isolates from normal dogs. The majority of isolates (171 of 249; 69% were located in two ETs (ET1 and ET 4), that were not distinguishable phenotypically. ET 1 contained 94 isolates (54 isolates from healthy dogs and 40 isolates from diseased sites) and ET 4 contained 77 isolates (46 from healthy dogs and 31 isolates from diseased sites). Further, 77.5% of isolates from healthy dogs were present in ET 1 and ET 4 and 59% of isolates from diseased dogs belonged to the same two ETs. There was only a small difference in genetic diversity among isolates taken from healthy dogs (11 ETs; H = 0.182) and those isolates taken from clinical specimens from diseased dogs (21 ETs; H = 0.218). Of the 21 ETs from diseased sites, ET 16 contained all six isolates from Staphylococcal Scalded Skin Syndrome in racing Greyhounds. CONCLUSIONS: The small difference in genetic diversity between isolates from the skin and mucosa of healthy dogs and isolates from various diseases, as well as the presence of the majority of isolates in two ETs, is consistent with the role of S intermedius as an opportunistic pathogen. The confinement of all Staphylococcal Scalded Skin Syndrome isolates within one ET is confirmation of this entity as a distinct disease of dogs. PMID- 10840576 TI - Occurrence of different strains of Dichelobacter nodosus in new clinical lesions in sheep exposed to footrot associated with multi-strain infections. AB - OBJECTIVE: To investigate the occurrence of S1, U1 and T strains of Dichelobacter nodosus in new clinical lesions in sheep exposed to footrot associated with multi strain infections. DESIGN: Seventy-seven donor sheep were grazed with 84 recipients for 33 weeks. The donor sheep were Merinos with a history of clinically virulent footrot associated with protease type S1, U1 and T strains of D nodosus that hybridised with gene sequences pJIR314B, pJIR318 and/or pB645-335. The recipient sheep were Merinos with no history of footrot. PROCEDURE: Each fortnight, all feet were examined, their lesion scores were recorded and samples of lesion material were taken for laboratory tests. RESULTS: Eighty-nine percent (299 of 336) of feet of recipient sheep developed new clinical lesions. S1, U1 and T strains of D nodosus were recovered from 58%, 22% and 18%, respectively, of these lesions at a ratio that remained constant during two apparent peaks in footrot transmission. Gene sequences homologous to pJIR314B and pB645-335 were detected in 56% (93 of 166) and 29% (48 of 166), respectively, of S1 strains of D nodosus at a ratio that was not constant during the experiment. CONCLUSIONS: S1 was the dominant protease type of D nodosus in new clinical lesions. The occurrence of S1 strains did not increase relative to U1 and T strains of D nodosus during the experiment. S1, U1 and T strains of D nodosus remained in equilibrium despite changes in environment, genetic types in the population of S1 strains, and host resistance to footrot. PMID- 10840577 TI - Incidence of equine herpesvirus 1 infection in thoroughbred weanlings on two stud farms. PMID- 10840578 TI - A fatal case of Hendra virus infection in a horse in north Queensland: clinical and epidemiological features. PMID- 10840580 TI - Effects of spinach powder fat-soluble extract on proliferation of human gastric adenocarcinoma cells. AB - Four kinds of assays were used to study the effect of a fat-soluble extract of spinach powder (SPFE) on the proliferation of human gastric adenocarcinoma cell line (SGC-7901) in vitro. These studies included: (i) cell growth assay, (ii) colony forming assay, (iii) MTT colorimetric assay, and (iv) 3H-TdR incorporation assay. The concentrations of SPFE expressed as the level of beta-carotene in the medium were 2 x 10(-8), 2 x 10(-7) and 2 x 10(-6) mol/L beta-carotene in assay (i)-(iii), but 4 x 10(-8), 4 x 10(-7) and 4 x 10(-6) mol/L beta-carotene in assay (iv) respectively. The results indicated that SPFE inhibited the proliferation and colony forming ability of SGC-7901 cells. And in MTT assay, SPFE inhibited the viability of SGC-7901 cells, but no inhibitory effect of SPFE was observed on the viability of lymphocytes in peripheral blood of healthy people. Finally, in the 3H-TdR incorporation test, both SPFE and beta-carotene showed significant inhibitory effects on DNA synthesis in SGC-7901 cells, but SPFE was more effective than beta-carotene. PMID- 10840579 TI - Identification and molecular characterization of Hendra virus in a horse in Queensland. PMID- 10840581 TI - Preliminary validation of tumor cell attachment inhibition assay for developmental toxicants with mouse S180 cells. AB - This study was designed to explore the possibility of using ascitic mouse sarcoma cell line (S180) to validate the mouse tumor cell attachment assay for developmental toxicants, and to test the inhibitory effects of various developmental toxicants. The results showed that 2 of 3 developmental toxicants under consideration, sodium pentobarbital and ethanol, significantly inhibited S180 cells attachment to Concanavalin A-coated surfaces. Inhibition was dependent on concentration, and the IC50 (the concentration that reduced attachment by 50%), of these 2 chemicals was 1.2 x 10(-3) mol/L and 1.0 mol/L, respectively. Another developmental toxicant, hydrocortisone, did not show inhibitory activity. Two non-developmental toxicants, sodium chloride and glycine were also tested and these did not decrease attachment rates. The main results reported here were generally similar to those obtained with ascitic mouse ovarian tumor cells as a model. Therefore, this study added further evidence to the conclusion that cell specificity does not limit attachment inhibition to Con A-coated surfaces, so S180 cell may serve as an alternative cell model, especially when other cell lines are unavailable. Furthermore, after optimal validation, it can be suggested that an S180 cell attachment assay may be a candidate for a series of assays to detect developmental toxicants. PMID- 10840582 TI - Application of YLD calculation in assessing disease data--an analysis of 4 diseases in 2 regions. AB - The objective of the current study is to discuss the problems related to how data is used to calculate Years Lost with Disability (YLD) with the method recommended by the World Bank. The study includes collecting useful data, estimating disease duration and average age of disease onset, adjusting incidence and prevalence data by means of a software programme, DISMOD (Harvard University Incidence & Prevalence Model), and assessing the importance of YLD calculation for different diseases. Remission and fatality rates of 3 diseases were estimated by experts at 2 round consultations. Incidence rates, disease duration and average age of disease onset were calculated and adjusted by DISMOD. YLD due to schizophrenia is the highest among 4 diseases in two regions. YLD is 18.88% in disability adjusted life year for 4 diseases in Xiacheng District, and 19.97% in Fuyang County. Available data can be used for the calculation of YLD after being adjusted. DISMOD is a useful instrument to test the internal consistency of incidence, prevalence, remission and fatality rate. The adjusted data are acceptable to experts and DISMOD. To get rational remission and fatality rates, we can use a cohort method through expert consultations. To reflect overall burden of disease, YLD calculation should be used. PMID- 10840583 TI - Urban-rural comparison of nutrient intake by adult women in Shaanxi Province, China. AB - Triplet surveys were conducted in the city of Xi' an and two villages (one in the vicinity and the other at a distance) in Shaanxi Province in China in October November (when agricultural activities were low), 1997, to elucidate nutrient intakes with a focus on possible urban-rural differences. Total food duplicate samples were collected from non-smoking and non-habitually drinking adult healthy women (about 50 subjects per site and 149 in total). The nutrient intakes were estimated from the weight of food items in reference to national food composition tables. On average, the women took 1873 kcal energy, 54 g protein and 37 g lipid per day, with a lipid energy ratio of 18.4%. Both excess and insufficient energy intake was observed as a result of food intake analysis and body mass index determination. With regard to minor nutrient intakes, insufficiency was serious in the case of calcium, vitamin A and vitamin B2, but not with iron. Whereas dependency on plant foods for sources of energy and protein was common to the three regions, Xi' an people consumed more animal foods than those in the villages. Intake of fish and shellfish was quite low throughout the three regions. Among the four types of cereals, wheat was consumed most substantially in the three regions and in three meals (except for the village where people essentially did not take lunch in reflection of low agricultural activities), whereas rice was consumed more in Xi' an than in the two villages. Maize consumption was higher in the two villages (especially for breakfast) than in the city. In contrast, foxtail millet (although in small amounts) was taken primarily in Xi' an and only at the time of breakfast. PMID- 10840584 TI - Influence of combined therapeutic potential of meso 2,3-dimercaptosuccinic acid and calcium disodium edetate on lead-induced testicular alterations in rats. AB - The therapeutic efficacy of a combination of meso 2,3-dimercaptosuccinic acid (DMSA) and calcium disodium EDTA in protecting testicular disorders in chronic lead intoxication was investigated. The results indicate that two five-days courses of the combined therapy produced a more effective recovery in the lead induced biochemical and histopathological disorders compared to conventional single 5 days therapy. No adverse effect of the chelators, when administered individually or in combination, was noticed in the testes of control (without lead exposure) animals. PMID- 10840585 TI - Induction of chromosomal aberrations by propoxur in mouse bone marrow cells. AB - Propoxur is a widely used dithiocarbamate insecticide. In this study, the clastogenic effect of propoxur has been evaluated using chromosomal aberration assay in mouse bone marrow cells. Single i.p. administration of propoxur, at 25 mg/kg b.wt., a maximum tolerated dose (MTD) and 12.5 mg/kg b.wt (50% of MTD) have significantly induced different types of aberrations after 24 h of treatment. The aberrations were dose and time dependent and reached a maximum after 24 h of exposure. The results suggest a genotoxic potential of propoxur. PMID- 10840586 TI - Preparation and immunogenicity of serogroup B meningococcal OS-OMPC conjugates. AB - The objective of the present study is to purify oligosaccharides (OSs) from serogroup B meningococci (3407 or 542852) with common lipooligosaccharide (LOS) immunotypes prevalent in China and to successfully conjugate them to their outer membrane protein complexes (OMPCs). Conjugates possessing broad cross-reactivity with different serogroup B meningococci were obtained. Both LOSs from the above two strains were purified on a Sephadex G75 column and hydrolyzed with acetic acid, and then the pure OSs were obtained by using a Sephadex G50 chromatographic column. The pure OSs were conjugated to corresponding OMPCs by carbodiimide mediated condensation. Mice were immunized with the OS-OMPC conjugates to produce antibodies. Immunogenicities of the conjugates were tested by ELISA, bactericidal test and Western-blotting. The titers of antibodies against OS and LOS elicited by the two OS-OMPC conjugates increased respectively by 2.5 and 2 to 10 times as compared with those of antibodies against unconjugated OS and LOS. The sera elicited by the conjugates possessed a stronger bactericidal activity to the strains (3407, 542852, 29021) with the same LOS immunotype than to the strain with different LOS immunotype (86155). The results determined by ELISA indicated that the sera against the conjugates showed strong response to the strains with the same or different LOS immunotypes. It was confirmed by western-blotting analysis that the sera elicited by the two conjugates reacted with 42, 39 and 26 kDa proteins from the OMPC. Among the reactive bands, the 42 kDa proteins were class 1 OMPs. The two serogroup B meningococcal OMPCs were strong immunogens and good carriers coupling with OSs as well. The immunogenicity of the conjugated OS was increased and the OS-OMPC conjugates of the serogroup B meningococci showed a broad antigenic response to the strains tested. PMID- 10840587 TI - The need of a physiologic and pathophysiologic definition of stress. PMID- 10840588 TI - Individual reactivity and physiology of the stress response. AB - The biological response to stress involves the activation of two main neuroendocrine components, the hypothalamic-pituitary-adrenal axis and the sympathoadrenal medullary systems. Looking at the responses to stressors in a laboratory setting, e.g., cortisol production after exercising on a treadmill, is a valid and controlled way to study how people react to psychological and physical stressors. A common finding in such studies is that individuals respond bimodally to stress. More recently, researchers have been interested in the possible reasons why healthy individuals exhibit differential reactivity to stressors. The literature on the neuroendocrine responses to stress, with a particular focus on investigations of individual reactivity to psychological and physical stressors, is reviewed. PMID- 10840589 TI - Melanocortins and feeding behavior. AB - The melanocortin (ACTH/MSH) peptides exert a number of central effects. In the eighties, we described for the first time a role for melanocortins in the central control of appetite. We showed that the injection of ACTH-(1-24) into a brain lateral ventricle reduced food intake up to 76.6% in starved rats. Injections into the ventromedial hypothalamus during the nocturnal feeding phase also markedly inhibited food intake. These effects were also confirmed in mice and rabbits. Targeted disruption of the MC4 receptor resulting in obesity in mice explained the role of this receptor in mediating effects of melanocortins on food intake. Administration of MC4 receptor agonists leads to acute reduction in food intake and body weight, while the reverse effects are observed after administration of selective MC4 receptor antagonists, confirming the role of the melanocortins in mediating a tonic inhibition on feeding behavior. Moreover, immobilization stress-induced anorexia may be partially reversed by single and repeated intracerebroventricular administration of selective MC4 receptor antagonists. It is thus evident that MC4 receptor blockage can reduce stress induced anorexia and that repeated injections of selective MC4 receptor antagonists have a sustained effect on food intake without any sign of tachyphylaxis. However, we have also shown that the behavioral effects of CRF (anorexia and grooming) are not influenced by MC4 receptor blockage. These effects of CRF are thus not due to an indirect mechanism caused by an increased release of melanocortins acting on the central MC receptors. PMID- 10840590 TI - Environmental stress and psychiatric illness. AB - It has long been recognized that environmental stress plays a pivotal role in the pathogenesis of psychiatric disorders. The relationship is complex and the neurobiological mechanisms that mediate the contribution of stressful experiences to the manifestation of illness are not well understood. In considering this relationship, it is important to differentiate between the role of environmental stressors as vulnerability factors that predispose the individual to psychiatric illness and may be temporally distant from its clinical onset, and their role as direct precipitants of the illness. Furthermore, environmental stressors must be considered in the context of constitutional vulnerability factors, such as genetic predisposition, with which such stressors may interact. Genetic predisposition may influence not only vulnerability to illness but also the nature of the individual's response to stress and the likelihood of exposure to stressful events. In this paper, we focus on two areas that illustrate the complexity of the field and the important findings that have emerged--the role of early parental loss (EPL) in adult psychopathology, particularly major depression, and the relationship between recent significant life events and depressive episodes. We conclude with a preliminary conceptual framework for considering the relationship between genetic susceptibility and environmental stress in the pathogenesis of psychiatric illness. PMID- 10840591 TI - Stress system alterations and mood disorders in suicidal patients. A review. AB - Stress system alterations, in particular HPA axis hyperactivations, are fairly well documented in suicide victims and in depressed suicide attempters who subsequently complete suicide. In suicide attempters with no documented completed suicide, the results are more inconsistent. This may depend on differences in studies due to diagnostic heterogeneity among suicide attempters, and the time between the suicide attempt and the examination. Recent data suggests differences in the stress system between depressed patients with a recent suicide attempt and depressed patients in general. The field merits further examination, with thorough examinations of genotypes, actual suicide attempts and stress in combination with examinations of the stress system. PMID- 10840592 TI - On the track of a human circulating mesenchymal stem cell of neural crest origin. AB - The neural markers present in the normal circulating monocytoid cells able, in pathological situations, to trans-differentiate into different mesenchymal-type cells, confirm the hypothesis previously raised that these cells derive from the neural crest. In culture, the normal cells display a great plasticity very reminiscent of microglial cells in culture. Almost a quiescent cell in normal individuals, this monocytoid cell shows its division potentialities in pathological situations of fibrosis and cancer (chondrosarcoma) where it is found to spontaneously proliferate. While the normal neofibroblasts are rapidly recognized and destroyed by fibrophagic T-lymphocytes, the pathological cells escape this control and, as a result, they accumulate in vitro giving rise to a tissue sometimes organized as nodules. Although basically the transdifferentiation process is similar in all the pathological situations of fibrosis and cancer studied so far, the end-result phenotype evokes the pathology the patient is suffering from. It evokes osteoblasts in a case of osteomyelosclerosis, chondroidocytes in a case of chondrosarcoma, myelofibroblasts in a case of fibrosis of lung and kidney in a patient under ciclosporine treatment. Hence, this circulating monocytoid cell is a multipotent cell with great division potentiality. These are characteristics of stem/preprogenitor cells. Since this circulating monocytoid cell also bears the neural markers we called it a monocytoid ectomesenchymal stem/preprogenitor cell. Therefore, the existence of an ectomesenchymal system is discussed here. The circulating monocytoid ectomesenchymal stem/preprogenitor cell might be involved in the normal cicatrisation process while the fibrophagic T lymphocytes might be involved in its termination. Impairment of this controlled mechanism might result in the development of fibrosis and/or cancer such as chondrosarcoma in vivo. Interestingly, at least in vitro, proliferation is restricted to the monocytoid cell before transdifferentiation takes place. In this model, fibrosis and cancer might share some common steps going from the proliferation of the monocytoid cells to their transdifferentiation into mesenchymal-type cells and the accumulation of these transdifferentiated cells in the tissues. Then, cancer might be distinguished from fibrosis by the additional acquisition of the ability to proliferate by the transdifferentiated cells. The monocytoid ectomesenchymal stem/preprogenitor cell might also be involved in brain neurodegenerative diseases characterized by an accumulation of microglia. The circulating monocytoid ectomesenchymal stem/preprogenitor cell appears as a target for gene therapy in pathological situations of fibrosis and/or cancer where it proliferates out of control. If the normal cell can be expanded and if its transdifferentiation can be directed, the circulating monocytoid ectomesenchymal stem/preprogenitor cell may become a useful tool for cellular therapy, in case of failure in wound healing and tissue regeneration. PMID- 10840593 TI - HBsAg as the antigen component of circulating immune complexes in HIV-infected patients. AB - Seeing the same transmission pattern of HIV and HBV coinfection by these two agents is not an uncommon feature. Immunity impairment due to HIV infection can be the cause of a higher rate of HBV replication with less intensive liver damage and less effective immune response to HBV, while the pathological course in both infections involves elevated levels of circulating immune complexes (CIC). These were the reasons for us to examine the frequency of HBsAg involvement as the antigen component of circulating immune complexes formed in sera of HIV-infected patients in different stages of HIV disease. We tested 67 sera of HIV-positive patients in different stages of HIV disease for the presence of HBsAg and HIV antigen p24 (with and without acid dissociation of immune complexes), for the presence of anti-Hbc antibodies and circulating immune complexes. HBsAg was positive in 13.8% sera prior to and 33.8% after acid pretreatment. Anti-HBc antibodies were present in 76.9% serum samples tested. Fifty percent of sera were positive for both HBsAg and p24 antigen after dissociation of immune complexes. The level of CIC was elevated in 65.9% of sera. Our results suggest that HBsAg is commonly associated in immune complexes formed in the sera of HIV-infected patients and that they may simultaneously contain HIV and HBsAg in patients coinfected with both agents. This may contribute to their mutual interaction and influence the diagnosis and follow-up of patients. PMID- 10840594 TI - Non-steroidal anti-inflammatory drugs might also be pro-inflammatory by increasing tumor necrosis factor. PMID- 10840595 TI - Anion exchange purification of plasmid DNA using expanded bed adsorption. AB - Recent developments in gene therapy with non-viral vectors and DNA vaccination have increased the demand for large amounts of pharmaceutical-grade plasmid DNA. The high viscosity of process streams is of major concern in the purification of plasmids, since it can cause high back pressures in column operations, thus limiting the throughput. In order to avoid these high back pressures, expanded bed anion exchange chromatography was evaluated as an alternative to fixed bed chromatography. A Streamline 25 column filled with 100 ml of Streamline QXL media, was equilibrated with 0.5 M NaCl in TE (10 mM Tris, 1 mM EDTA, pH = 8.0) buffer at an upward flow of 300 cmh-1, E. coli lysates (obtained from up to 3 liters of fermentation broth) were injected in the column. After washing out the unbound material, the media was allowed to sediment and the plasmid was eluted with 1 M NaCl in TE buffer at a downward flow of 120 cmh-1. Purification factors of 36 +/- 1 fold, 26 +/- 0.4 plasmid purity, and close to 100% yields were obtained when less than one settled column volume of plasmid feed was injected. However, both recovery yield and purity abruptly decreased when larger amounts were processed-values of 35 +/- 2 and 5 +/- 0.7 were obtained for the recovery yield and purity, respectively, when 250 ml of feedstock were processed. In these cases, gel clogging and expansion collapse were observed. The processing of larger volumes, thus larger plasmid quantities, was only possible by performing an isopropanol precipitation step prior to the chromatographic step. This step led to an enhancement of the purification step. PMID- 10840596 TI - Bovine serum albumin-hemoglobin fractionation: significance of ultrafiltration system and feed solution characteristics. AB - This work investigates the fractionation of similar molecular weight proteins bovine serum albumin (69 kD) and bovine hemoglobin (67 kD) by ultrafiltration. Three different membranes, viz. regenerated cellulose, poly(sulfone) and surface modified poly(acrylonitrile), each with a nominal molecular cutoff rating of 100 kD, were examined. The experiments were conducted in dead end, crossflow and vortex flow filtration modes and the separation was studied as a function of feed pH and ionic strength. Under similar system hydrodynamics, the surface modified poly(acrylonitrile) membrane displayed the highest resolution with minimum membrane fouling. The separation could be improved further by operating at low applied pressure (40 kPa) and high mass transfer (> 20 x 10(-6) m/s) in a vortex flow module. Under these conditions, the highest separation factor of 40 was obtained at the pI of hemoglobin. PMID- 10840597 TI - Expanded bed chromatography of proteins in small diameter columns. I. Scale down and validation. AB - The use of large columns for expanded-bed chromatography in protein adsorption and purification can pose limitations in method scouting due to the high volumes of consumables involved in optimisation runs. Scaling down this technique would provide a practical and necessary first step for demonstrating its feasibility in very small beds. The performance of three columns of diameters 5.0, 1.0 and 0.5 cm were compared in terms of the bed expansion, hydrodynamics and breakthrough for lysozyme adsorption onto STREAMLINE-SP. This represented a scale-down factor of a 100-fold from the 5-cm column and the success was judged by the insignificant changes in performance based on the selected criteria. Bed characterisation and breakthrough runs indicated good plug flow behaviour, despite the high particle size to column diameter ratio in the smaller columns. The column efficiency was found to be sensitive to the vertical alignment, making it an important issue in scale down. The results of these investigations show that small diameter columns can be effectively used for mimicking the behaviour in scale up systems providing a useful tool for method scouting studies. PMID- 10840598 TI - Expanded bed chromatography of proteins in small-diameter columns. II. Methods development and scale up. AB - The scaled down system developed in Part I of this series was further validated by using a 1-cm diameter column for method development studies for the separation of two model proteins, alcohol dehydrogenase and alpha-glucosidase, from unclarified yeast homogenate by hydrophobic interaction expanded bed chromatography based on the STREAMLINE matrix. The efficacy of solids removal and establishment of optimal binding and separation condition by stepwise elution were investigated. Equilibration of the EBA column and loading at high salt strengths affected the subsequent recovery of the two target proteins. Although good resolution between the target proteins could be achieved, peak tailing was found to be a consistent problem. The optimised separation protocol was scaled up 25-fold to a column diameter of 5.0 cm. The results were in good agreement with the run conducted in the 1-cm column, indicating the potential of using the small columns as an viable approach for method scouting and development studies. PMID- 10840599 TI - Contamination of an anion-exchange membrane by glutathione. AB - Electrodialysis, which can separate electrolytes under mild conditions by using ion-exchange membranes, is a strong candidate for separation of GSH from yeast extracts, because GSH is unstable and easily oxidized forming a disulfide bond especially under alkali conditions. In this paper, sorption behavior of GSH on an anion-exchange membrane, in the pH 3-6 region that is expected to be the most preferable for its electro-dialytic separation, was examined. Sorption of GSH on a Selemion-AMV anion-exchange membrane was accelerated as the pH of the membrane contact solution increased, and there was a good correlation between the sorbed amounts and the molar fraction of monovalent anionic species of GSH. However, the amounts of GSH desorbed from the membrane by a NaCl desorbing solution were much lower than the initial sorbed amounts, and the difference between them was enlarged with increasing pH. The GSH which was lost could be recovered by the addition of DTT in the membrane-contact and desorbing solutions. Similar results were also obtained with Cys. We thus concluded that an anion-exchange membrane would be contaminated by thiol compounds, such as GSH and Cys, through oxidative binding of the thiol group with the membrane, the local OH- concentration in which was enhanced due to attraction by the positively charged anion-exchange membrane. PMID- 10840600 TI - A new method for yeast recovery in batch ethanol fermentations: filter aid filtration followed by separation of yeast from filter aid using hydrocyclones. AB - In the Melle-Boinot process for alcohol production, centrifuges are normally used for yeast recovery at the end of a batch fermentation. Centrifuges are expensive equipment and represent an impressive part of the equipment costs in alcohol industries. In the present work, an alternative method for yeast recovery using less expensive equipment was studied. Instead of using centrifuges, yeast was separated from the fermented broth by filter aid filtration, followed by separation of yeast from the filter aid using hydrocyclones. A stainless steel plate-and-frame filter of filtration area 1.14 m2 and two 30 mm hydrocyclones, which followed the Bradley and Rietema recommended proportions, were used in this work. The filter aid was perlite. Tests of direct separation of yeast from the fermented broth using the Bradley hydrocyclone proved to be completely unfeasible, since the maximal reduced total efficiency obtained was only 1%. When the hydrocyclones were used to separate perlite from the resuspended filtration cake, the perlite total separation efficiency obtained in the underflow was as high as 95% when using the Bradley hydrocyclone with an underflow diameter of 3 mm. To show the feasibility of the proposed new method of yeast recovery, a complete cycle of experiments, which included fermentation, yeast separation, and new fermentation using the recycled cells, was performed with good results. PMID- 10840601 TI - [Diagnosis and therapy of abnormal bleeding in surgery]. AB - Abnormal surgical bleeding may be caused by congenital or acquired plasmatic and/or thrombocytic coagulation disorders. They may be known at the time of operation, or they may occur during the perioperative course for the first time. A detailed identification of haemostatic defects can be performed by a diagnostic spectrum of multiple tests. However, in clinical practice global screening tests are used first. In diagnostic routine three groups of patients have to be considered: (1) patients with no personal or family history of bleeding and no operative bleeding risk; (2) patients with no history, but increased bleeding risk by the planned procedure; (3) patients with a known haemorrhagic diathesis in their own history. In all difficult situations a specialist in haemostasiology should be consulted, because only rational diagnosis and therapeutic monitoring achieve an optimal and cost-effective operative result. The surgeon should always be aware that surgical bleeding from an operative cause must be considered in the differential diagnosis. PMID- 10840602 TI - [Bleeding problems in liver surgery and liver transplantation]. AB - In liver resection, severe bleeding can be prevented by appropriate surgical techniques. These include adequate access and mobilisation, vascular occlusion, controlled dissection of the parenchyma, prevention of venous "over-filling" and secure hemostasis of the resection surface. Excessive bleeding, both in liver resection and liver transplantation, poses a major risk for the development of postoperative complications. In liver transplantation the surgeon is most often confronted with patients with chronic liver disease and portal hypertension. Coagulation disorders are the rule, and the surgery itself is more demanding because of fragile venous collaterals as a consequence of portal hypertension. With the use of extracorporeal venovenous bypass or newer techniques with preservation of the vena cava, some of these difficulties can be overcome. Pharmacological therapies like administration of aprotinin can reduce the fibrinolysis inherent in liver transplantation. However, surgical skill and experience are probably still the most important predictors of blood loss during surgery. PMID- 10840603 TI - [Bleeding problems in open heart surgery]. AB - Across centers, transfusion can vary eightfold for the same mediastinal drainage after adult cardiac operations. Excessive blood drainage resulting in increased transfusions occurs in 29% of patients. Various strategies have been proposed to decrease bleeding and allogenic transfusion requirements in the perioperative period of heart surgery. Blood conservation methods were reviewed critically. Avoidance of preoperative anemia, tolerance of low hemoglobin concentrations, the use of autologous blood, and adherence to a strict transfusion protocol will reduce the use of allogenic transfusions. Perioperatively, maintenance of normothermia contributes to improved hemostasis. PMID- 10840604 TI - [Epidemiology, diagnosis and prevention of viral hepatitis]. AB - Viral hepatitis is one of the most important infectious diseases worldwide. It is frequent in tropical and subtropical areas, but also plays still a significant role in the industrialized countries of Northern and Western Europe and the USA. In Germany more than 100,000 people are infected by viral hepatitis each year. Whereas hepatitis A and E are transmitted fecal-orally, hepatitis B, C and D are transmitted by the parenteral route. Most hepatitis A infections seen in Germany are acquired abroad. Hepatitis B is still a nosocomial disease; the majority of infections in Germany, however, seems to be transmitted sexually. For the serological diagnosis of viral hepatitis a battery of sensitive tests for specific antibodies and antigens is available; usually testing of one serum sample allows an unambiguous diagnosis. Prophylaxis of hepatitis A and B is possible using safe and efficient vaccines. PMID- 10840605 TI - [Clinical aspects and therapy of viral hepatitis]. AB - Acute hepatitis can be caused by the enterically spread hepatitis A and E viruses and the parenterally spread hepatitis B, C or D viruses. The clinical features of acute viral hepatitis are similar among the five viruses and include non-specific symptoms and icterus. In general, a specific therapy is not necessary, but patients with fulminant hepatitis may require liver transplantation. For acute hepatitis C, the effect of interferon-alpha on the risk of chronicity is evaluated in clinical trials. Chronic hepatitis is defined as inflammatory reaction in the liver that continues without improvement for at least 6 months after infection with hepatitis B, C or D viruses. Hepatitis B resolves in more than 90% of the patients, but chronic infection can lead to liver cirrhosis and hepatocellular carcinoma. Chronic hepatitis C is an insidious disease, because early diagnosis is missed easily due to asymptomatic presentation and about 70% of infected patients develop chronic hepatitis. The benefits of interferon-alpha and/or nucleoside analogues have been proven in recent clinical trials that show sustained responses in more than a third of all patients with chronic viral hepatitis. The future treatment of chronic viral hepatitis will likely include immunomodulation and gene therapy. PMID- 10840606 TI - [Transmission of HBV, HCV and HIV by infectious medical personnel--presentation of an overview]. AB - This report analyzes 46 cases of personnel-to-patient transmissions of hepatitis B (HB), hepatitis C (HC) and HIV in health care settings. Similar circumstances were found for transmission of HB (40 cases, 404 infected patients), HC (4 cases, 224 infected patients) and HIV (2 cases, 7 infected patients). Cases with the highest number of transmissions (one anesthesiologist with 217 HC transmissions, and one EEG technologist with 75 HB transmissions) were attributed to poor infection control practices. As long as infected health care workers (HCW) adhere to general infection control measures, a risk for transmission to patients exists only from infected surgeons who perform 'exposure-prone invasive procedures'. Whether changes in duties of infected HCW are necessary should be decided on an individual basis. Often, the infected personnel were assumed to have acquired the disease occupationally. Medical practices and devices bearing a risk of infection should constantly be reviewed with regard to risk for patients and personnel. PMID- 10840607 TI - [European and North American regulations on employing HBV-, HCV- and HIV-infected persons in health care]. AB - In the United States of America (1991), in Canada (1998), in the UK (1993, 1994) and in Germany (1999) recommendations have been published concerning performance of "exposure-prone procedures" (EPP) by persons infected with Hepatitis-B virus (HBV), Hepatitis-C virus (HCV) or HIV. This review summarises the definition of EPP, methods to reduce the transmission of blood-borne pathogens, immunisation and screening policies, constitution of an advisory panel and the question of trace-back investigations described in these regulations. PMID- 10840608 TI - [Hepatitis and liver transplantation]. AB - Chronic hepatic dysfunction and acute liver failure due to viral hepatitis represent the most frequent indications for liver transplantation. Liver grafting is the only available life-saving intervention for most of these patients. Reinfection of the graft is still a central problem. In hepatitis B, reinfection frequently leads to cirrhosis with subsequent dysfunction of the graft. Prophylaxis of HBV reinfection with polyclonal human antibodies against HBsAg alone or in combination with the nucleoside analogue lamivudine is effective but costly. Reinfection in hepatitis C is a regular event in all patients and has a far better prognosis than HBV reinfection of the graft. Effective prophylaxis of HCV reinfection has still not been established and requires further studies. After HCV reinfection the prognosis of allograft survival is negatively affected; however, the 5-year survival rate in patients who have undergone liver transplantation for HCV-related liver disease is not lower than for other non viral benign indications. PMID- 10840609 TI - [Detection of isolated disseminated tumor cells of colorectal carcinomas in lymph nodes]. AB - The 5-year survival of patients with colorectal cancer UICC stage I and II ranges from 70% to 80%. Therefore, at least 20% to 30% of these patients develop recurrent disease, caused by hematogenic or lymphogenic tumor cell dissemination. Using immunohistochemical or molecular biological methods, disseminated tumor cells can be detected in lymph nodes in a high proportion of patients in UICC stage I or II. The detection of disseminated tumor cells could help to identify a subgroup of patients at risk for disease relapse who could benefit from adjuvant therapy. Lymphogenic tumor cell dissemination seems to be a very common event in early colorectal cancer, preceding hematogenic tumor cell dissemination. Although these observations appear to confirm the therapeutic importance of meticulous lymph node dissection, their prognostic relevance has yet to be determined in prospective studies. PMID- 10840610 TI - [Health care reform 2000--effects on inpatient care]. AB - The health reform 2000 in Germany has little impact on the structural deficits of the German health service. The monopolies of hospitals for inpatient and of contract doctors for outpatient care are not changed. The budgeting of hospitals remains. In future there will be a catalog of operations that may only be carried out on an outpatient basis. The duty for quality assurance is specified and intensified. Implementation of innovative medical treatments in hospital will to be done only under strict control of a new countrywide committee. A new DRG-based price system for hospitals will be established. PMID- 10840611 TI - [Current status of the future surgical continuing education structure]. PMID- 10840612 TI - [Serous cystadenoma of the pancreas]. AB - INTRODUCTION: Serous cystadenomas of the pancreas are rare tumors and thought to be almost always benign. METHODS: We report our experience in the diagnosis and surgical treatment of 12 patients with these tumors. RESULTS: Between October 1993 and December 1998, 41 patients with cystic tumors of the pancreas underwent surgical resection; in 12 cases (11 women, 1 man) a serous cystadenoma (10 micro cystic, 2 oligo-microcystic) was found. Only 6 (50%) patients had symptoms. The mean tumor size was 4.8 (2.7-10) cm. Ultrasound, CT and MRT usually could detect the mass, but differentiation with other cystic lesions was not reliable. All tumors were resected: 4 Whipple procedures, 7 distal pancreatectomies and 1 segmental resection were performed. No patient died after surgery and none had to be reoperated on. CONCLUSIONS: Because of the difficulty in reliably differentiating benign and malignant lesions of the pancreas, we believe that cystic tumors of the pancreas should be resected. PMID- 10840613 TI - [Diagnosis and therapy of well-differentiated neuroendocrine lung tumors]. AB - INTRODUCTION: Neuroendocrine tumours of the lung are uncommon malignant neoplasms. They represent a heterogeneous spectrum of disease, encompassing the well-differentiated neuroendocrine tumours through to small-cell lung cancer, which has a poor prognosis and only palliative therapeutic options for most patients. Due to the low incidence of well-differentiated lung tumours there is as yet little experience. METHODS: Clinical records of 14 patients with well differentiated neuroendocrine tumours of the lung were assessed retrospectively for patients' initial symptomatology, diagnostic procedures, therapy and results. RESULTS: Eight lobectomies, 2 bronchoplastic lobectomies, 2 segmental resections, 1 bilobectomy and 1 wedge resection were performed; seven of them without regional lymphadenectomy. All patients were alive after a mean follow-up of 38 (2 96) months. Thirteen of 14 patients were free of complaints, twelve had no recurrences and returned to work. CONCLUSIONS: Well-differentiated NET of the lung have an excellent prognosis after curative resection. Limited resections even without lymphadenectomy can be performed in some cases. PMID- 10840614 TI - [Conventional aortic surgery after endovascular aortic reconstruction]. AB - INTRODUCTION: Endovascular therapy of aortic aneurysms is an attractive alternative to conventional aortic surgery. A number of patients, however, will fail endovascular therapy and require open surgical repair. METHODS: We report our experience with conversion to open aortic repair. Of 306 patients given endovascular therapy for an infrarenal aortic aneurysm, 30 (9.8%) required conversion to conventional repair. RESULTS: Thirteen patients (43.3%) suffered complications, 3 (10%) of which were fatal. Conversion to open aortic repair after endovascular therapy is technically more demanding than primary open repair, particularly because of clamp techniques and stent-extraction. CONCLUSIONS: Emergent conversion operations, especially when preceded by significant retroperitoneal bleeding from vascular tears due to endovascular manipulations are complicated by significant morbidity and mortality. PMID- 10840615 TI - [Hereditary colonic carcinoma without polyposis (HNPCC) without satisfying the Amsterdam criteria]. AB - Epidemiologic data suggest that an underlying genetic disposition can be detected in up to 10% of all colorectal cancer patients and autosomal dominantly inherited hereditary non-polyposis colorectal cancer (HNPCC) is the entity most frequently identified. It was described first by A. Warthin in 1895 in "Family G" and is characterized by a predisposition to an early onset of colorectal cancer and other intestinal or genitourinary tumors. We report the case of a 61-year-old woman with five different cancers. Although the strict Amsterdam Criteria were not fulfilled, molecular analysis revealed HNPCC; further genetic testing in the family confirmed that the 36-year-old and so far healthy son had inherited the germline mutation of his affected mother. Genetic testing in clinically suspected HNPCC cases is recommended for patients with colorectal cancer meeting the Amsterdam Criteria. In patients meeting one of Bethesda Criteria 2-7 without meeting the Amsterdam Criteria, germline mutation analysis is recommended only in MSI-positive tumors. PMID- 10840616 TI - [Mason vertical gastroplasty in treatment of morbid obesity. Results of a prospective clinical study]. AB - Morbid obesity (body mass index > 40 kg/m2) is a risk factor for cardiovascular, pulmonary, metabolic, neoplastic, and psychologic sequelae. In the present prospective clinical study 65 patients (11 men, 54 women) underwent vertical banded gastroplasty (Mason procedure) from June 1994 to October 1997. The median age was 41 +/- 5.3 years (range 18-69; n = 65). Preoperative body weight was 135 +/- 23 kg (96-229; n = 65), excess body weight in kg was 75 +/- 6.9 (44-155; n = 65) or in % 126 +/- 10 (78-223; n = 65) and BMI was 49 +/- 7.4 kg/m2 (39-69; n = 65). Mean hospital stay was 9.7 +/- 2.4 days (6-18; n = 65). Hospital mortality was 0% (0/65). Early complications were vomiting (30%) and problems in wound healing (15%; n = 65). Late complications (> 30 days) were incisional hernias (13.8%) and staple-line disruptions (12.3%; n = 65) with a reoperation rate of 23% (15/65). Median follow-up was 15.0 +/- 5.2 months (2-42) with a follow up rate of 100%. Mean weight loss after 12 months was 38.5 +/- 17 kg (30-98; n = 34) (P < 0.0001) and loss of excessive body weight 65 +/- 10% (57-86; n = 34), respectively (P < 0.0001). Cardiovascular risk factors (hypertension, diabetes, hyperlipidemia) were significantly improved within 12 months (n = 34). Vertical banded gastroplasty (Mason procedure)--well established for 20 years--is a good, safe therapy for morbid obesity if strict indications for operation are observed and if there is multidisciplinary long-term follow-up. Comorbid risk factors are considerably reduced and a long-term weight loss of more than 50% can be achieved without the risk of pathological metabolic changes. PMID- 10840617 TI - [Laparoscopic latero-lateral pancreaticojejunostomy]. AB - Side-to-side pancreatojejunostomy for patients with chronic pancreatitis and dilated pancreatic duct is an established procedure, but so far there has been no report of this operation being performed as minimally invasive surgery. A 23-year old woman with a history of seven years of recurrent pancreatitis with pancreas divisum underwent a latero lateral pancreatojejunostomy with Braun anastomosis. For access, three ports were used; the anastomoses were produced with Endo-GIA staplers. The operation time was 115 min and the postoperative course was uneventful. During 8 months of follow-up no dilatation of the pancreatic duct or signs of recurrent pancreatitis were seen. The patient feels fine. Using improved and newly developed surgical devices (i.e., ultrasonic shears, endo-staplers) more difficult procedures can be done in a reasonable operation time. Under ideal conditions, patients will have full benefits with minimally invasive surgery. PMID- 10840618 TI - [Laparoscopic closure of a Bochdalek hernia in the adult]. AB - We report the case of a 27-year-old female patient with a symptomatic Bochdalek hernia operated on laparoscopically. We discuss the diagnostic strategy and options in therapy. PMID- 10840619 TI - [Budd-Chiari syndrome--a rare manifestation of hereditary thrombophilia]. AB - Budd-Chiari syndrome is a rare manifestation of hereditary or acquired thrombophilia. We saw a case of Budd-Chiari syndrome in a 30-year-old woman leading to initial diagnostic difficulties. She underwent surgical side-to-side shunt and 9 weeks later an almost normal liver could be demonstrated on computerized tomography. Budd-Chiari syndrome should be considered if the Chiari triad with abdominal pain, hepatomegaly and ascites occurs in a patient. If necessary, invasive diagnostic procedures (e.g. angiography) must be performed. Therapeutic options are anticoagulative therapy and porto-systemic shunt, either as a TIPS or a surgical shunt. If severe liver failure occurs or liver cirrhosis is present, orthotopic liver transplantation is an additional option which also cures hereditary thrombophilia. PMID- 10840620 TI - [The window in the falciform ligament. A rare case of hernia of the small intestine through the falciform ligament]. AB - The most common causes for small-bowel obstructions with subsequent strangulation are intra-abdominal adhesions and external hernias. We report an unusual case of strangulated small-bowel hernia through a congenital window in the falciform ligament. The diagnosis and surgical treatment are discussed. PMID- 10840621 TI - [Pseudoaneurysm of the pediatric popliteal artery after arthroscopic meniscus resection]. AB - Arterial injuries following arthroscopic procedures are extremely rare, but may have dramatic consequences without early diagnosis and appropriate treatment. We report a case of popliteal artery pseudo-aneurysm a following arthroscopic meniscectomy in a child. Non-invasive diagnostic workup included Doppler flow color imaging and computed tomography with contrast and 3D workup. The arterial lesion was repaired surgically by an posterior approach using a vein patch plasty. The popliteal artery is in close relationship to the posterior capsule of the knee joint. During knee flexion the vessel is positioned forward and placed even closer to the horn of the lateral meniscus. For this reason arthroscopic surgical manipulation in the posterior aspects of the knee joint must be performed under direct visualization. Indistinct swelling in the popliteal fossa and calf following arthroscopic surgery should arouse suspicion of an arterial injury. PMID- 10840622 TI - [Surgical specialty department in the world wide web. Tribute to contemporary life style or information network?]. AB - An increasing number of surgical departments is creating its own home-page in the World Wide Web (WWW). We investigated 184 presentations concerning their quality in content and technical realization. Our research revealed that most presentations regard patients as their main target group in order to meet the patients' demand for information and transparency. However, universities are especially concentrating on scientific development and neglecting the patients. Only a few presentations contain means for web-based training. In general, technical realization seems to be much better than the content itself. As patients tend to appear as customers who will choose the hospital they feel comfortable with, every clinical department that is a competitor should create a communication and information base for their customers. Thus, they will be able to keep in touch, present their treatment types and to enable patients to compare a multitude of surgical departments. The usefulness for colleagues or cooperating institutions must be improved. The WWW has become an important center of information in surgery. PMID- 10840623 TI - [Victor von Hacker (1852-1933)--recollections of the life and works of an important Billroth student]. AB - The year of 1999 marks the 75th anniversary of Viktor von Hacker's retirement as chairman of the Department of Surgery at Karl-Franzens University School of Medicine in Graz. He was a favored pupil of Theodor Billroth (1829-1894). When he took his professorship at Graz in 1903, the present hospital, then as now one of the largest in Europe, was still in planning, and he immediately involved himself in all the subsequent developmental stages. Now we are again living through a stage of major reconstruction and expansion within the context of the "LKH 2000" project which also encompasses the surgical department and wards, most of which have generally remained unchanged since Hacker's time. During his 21 years as chairman of the department, Hacker made major contributions in the fields of gastro-intestinal surgery, esophageal surgery and especially esophagoscopy, as well as plastic surgery. Towards the end of his career, 70 years ago, Viktor von Hacker was named an honorary member of the German Society of Surgery. For these reasons, the man and his work should be recalled. PMID- 10840624 TI - [Ankle joint fractures in adults]. PMID- 10840625 TI - [Computerized prognosis--Riyadh in medical and legal ethical evaluation. Legal aspects of prognosis and treatment responsibility]. PMID- 10840626 TI - The history of the Electroencephalography and Clinical Neuroscience Society (ECNS). Part I: A brief history of the American Medical Electroencephalographic Association (AMEEGA). PMID- 10840627 TI - The History of the Electroencephalography and Clinical Neuroscience Society (ECNS). Part II: The American Psychiatric Electrophysiology Association (APEA): history and mission. PMID- 10840628 TI - Somatosensory evoked spikes and epileptic seizures: a study of 385 cases. AB - We examined 385 children whose EEG showed high voltage potentials evoked by taps applied to one or both feet or hands (SES). The relationship between characteristics of SES and the occurrence of epileptic seizures and the characterization of epileptic syndromes were studied. Ninety-one children (23.6%) had epilepsy, 42 (10.9%) had only febrile convulsions and 252 children had other complaints. Epilepsy occurred in a higher proportion of cases when: SES by foot tapping were multiphasic, with high amplitude or SES were obtained by hand stimulation and there was spontaneous epileptiform activity in the EEG. The following epileptic syndromes were diagnosed: benign childhood epilepsy with centrotemporal spikes in 21 cases, benign epilepsy of childhood with occipital paroxysms in 2, benign psychomotor epilepsy in 1, "partial idiopathic others" in 43, generalized idiopathic in 8, symptomatic epilepsies in 13 and undetermined in 3 cases. In most cases SES were observed in children without evidence of cerebral organic lesion, suggesting the existence of an age-related, functional mechanism. Some characteristics of SES and the occurrence of spontaneous epileptiform activity showed a positive association with epileptic seizures. SES occurred in different types of partial and generalized epilepsies of childhood but in nearly 50% of the cases with epilepsy, there was a benign condition involving mainly the parietal lobe with versive, unilateral and sleep-generalized seizures. PMID- 10840629 TI - Electroclinical features in children and adolescents with epilepsy and/or migraine, and occipital epileptiform EEG abnormalities. AB - This study attempted to better define clinical and EEG features for differential diagnosis between epilepsy and migraine in children with occipital epileptiform EEG abnormalities. We studied 126 children (57 males, 69 females; age 4-18 years) suffering from epilepsy (63), migraine (43) or both (20). Patients were selected because of the presence of epileptiform abnormalities in the occipital regions on their EEG at rest. Differences among groups were statistically analyzed (Pearson chi square; ANOVA) for sex, age at onset of seizures and migrainous attacks, family history, ictal signs and symptoms, EEG at rest (unilateral vs bilateral distribution of epileptiform abnormalities), and EEG during Hyperventilation (HV) and Intermittent Photic Stimulation (IPS). Significant differences were found in family history, ictal signs and symptoms, EEG at rest and during activation tests. A family history of epilepsy, visual symptoms such as colored hallucinations and micro/macropsias, frequently associated with clinical signs in the visual system (eye deviation, nystagmus), unilateral EEG abnormalities, and abnormal response to IPS were closely related to diagnosis of epilepsy. On the other hand, a family history of migraine, visual symptoms such as amaurosis and scotomata, without evident clinical signs, bilateral EEG abnormalities, and no changes during IPS were significantly related to migraine. In conclusion, these clinical and EEG differences should be considered in the differential diagnosis between epilepsy and migraine in children with occipital epileptiform EEG abnormalities. PMID- 10840630 TI - Investigation of the cerebral response to flicker stimulation in patients with headache. AB - Migraine affects 10% of the population, yet there is no proven diagnostic test. To date the most promising neurophysiological diagnostic technique has been the analysis of cerebral responses to photic stimulation or flicker (the "H" response). This test has proved to be sensitive in patients with "classic" migraine, but has not been re-evaluated since the introduction of the 1988 International Headache Society (IHS) classification system. In 33 migraineurs, we found that the "H" response was sensitive (86.4%) and specific (97.5%) in those with migraine defined according to IHS Committee as type 1.2.1 (typical aura) and type 1.1 (without aura), but not in basilar migraine (1.2.4). PMID- 10840631 TI - Reduction of auditory P50 gating response in marihuana users: further supporting data. AB - This report attempts to replicate our recent finding of a significantly reduced sensory gating response in medically and psychiatrically normal chronic marihuana users. After exclusions, 10 normal heavy marihuana users (> or = 3 times per week) and 10 normal non-user controls were tested with the paired auditory P50 sensory gating procedure. Sensory gating ratios were significantly higher (i.e., impaired suppression) for THC users as compared to controls. Using combined data from the current and previous report, the degree of sensory gating impairment among THC users was significantly correlated with the frequency of marihuana use per week. Suggestions for further research are offered. PMID- 10840632 TI - The Mozart effect: distinctive aspects of the music--a clue to brain coding? AB - The goal of this study was to determine distinctive aspects of Mozart music that may explain the "Mozart Effect," specifically, the decrease in seizure activity. As many as 81 musical selections of Mozart, but also 67 of J.C. Bach, 67 of J.S. Bach, 39 of Chopin and 148 from 55 other composers were computer analyzed to quantify the music in search of any distinctive aspect and later to determine the degree to which a dominant periodicity could be found. Long-term periodicity (especially 10-60 sec, mean and median of 30 sec), was found often in Mozart music but also that of the two Bachs, significantly more often than the other composers and was especially absent in the control music that had no effect on epileptic activity in previous studies. Short-term periodicities were not significantly different between Mozart and the Bachs vs. the other composers. The conclusion is that one distinctive aspect of Mozart music is long-term periodicity that may well resonate within the cerebral cortex and also may be related to coding within the brain. PMID- 10840633 TI - Changes in functional coupling between neural networks in the brain during maturation revealed by omega complexity. AB - To study age-dependent changes in coupling between cortical neural networks we applied a new method (omega complexity) to determine overall coherence of EEGs of 34 subjects ranging in age from 3 months to 16 years. We found that the functional coupling between different brain regions is low at birth and increases significantly in the first two decades of life. We suggest that this coupling depends critically upon the system of associational and callosal fibers which is unmyelinated at birth, and which only finishes myelinization in the second or third decade. Thus age-dependant changes in omega complexity may reflect maturation of brain structures underlying higher cerebral functions. If these results can be replicated, preferably in prospective, cohort rather than transectional type studies, omega complexity might prove to be clinically useful as an objective, quantitative measure of brain maturation. PMID- 10840634 TI - Effects of scopolamine on interhemispheric EEG coherence in healthy subjects: analysis during rest and photic stimulation. AB - The present study of coherence analysis, in 16 healthy male volunteers, aged 24 31 years, showed that the administration of 0.25 mg of scopolamine significantly reduced interhemispheric coherence in the delta and beta-1 bands in the resting state. Scopolamine also caused a significant increase both in EEG coherence during PS and in PS-related coherence reactivity in the beta band. In addition, this compound significantly reduced total WMS scores. These findings suggest that, in addition to causing cognitive impairments, central cholinergic dysfunction can alter interhemispheric functional connectivity under both nonstimulus and stimulus conditions. PMID- 10840635 TI - Early detection of neurophysiological abnormalities in infection by human immunodeficiency virus. AB - The human immunodeficiency virus causes serious, progressive and irreversible deterioration of the immunocompetence system and of the nervous system, so neurological pathology in infected patients is frequent (30-40%), affecting both the central and the peripheral nervous systems. There are different clinical and laboratory indicators of bad prognosis, considering the important neurotropism of the virus. This study attempts to evaluate which neurophysiological parameters are altered during the initial phases of infection by HIV. A total of 46 individuals were studied, 30 seronegative and 16 seropositive in stage A of the CDC-93 classification. Motor and sensory conduction studies were carried out on all of them on the upper and lower extremities, as well as visual, somatosensory and auditory evoked potentials and endogenous potentials, mainly P300. The analysis of the neurophysiological parameters evaluated in our series, showed alterations of the conduction velocity of the sural nerve, latency of N1 of the SSEP of median and posterior tibial nerves and P300 in the initial phases of the infection even in the absence of clinical symptomatology. PMID- 10840636 TI - Spectroscopic properties of a novel neutral proteinase from Saccharomonospora canescens. AB - A neutral proteinase (NPS) was purified from the culture broth of Saccharomonospora canescens sp. novus, strain 5, using DEAE cellulose and a POROS HQ/M 4.6 x 100 mm column. The stability towards thermal and chemical (guanidine hydrochloride, Gdn.HCl) denaturation of NPS was investigated by kinetic and equilibrium studies. The unfolding processes were monitored by circular dichroism and fluorescence spectroscopy. The free energy of stabilization in water was calculated to be 2.1 kcal mol-1. The thermostability was determined by the critical temperature Tc from fluorescence measurements (69 degrees C) and the melting temperature Tm (70 degrees C) from (1) measurements. Quenching with acrylamide, iodide and cesium gives information about the microenvironment of intrinsic protein fluorophores. The Ksv constant for NPS is 4.6 and classifies the emitting tryptophans as 'buried' in the hydrophobic interior of the investigated protein. PMID- 10840637 TI - Correlations between cadmium treatment, oxygen uptake and metallothionein response in liver and kidney from two mice strains. AB - Oxygen uptake was determined in liver and kidney slices from Balb/C and C3H/HeJ mice at various time points related to a 4-day cadmium (Cd) i.p. injection regime, the latter strain being not only the higher metallothionein (MT) responder, but also more Cd sensitive. In both strains Cd injections caused an increase in O2 uptake in the tissues, being more expressed in the higher MT responsive C3H/HeJ strain. An ability for fast, additional increase of O2 uptake in response to subsequent Cd exposure was also induced during the injection regime. Maximal obtainable MT levels in the two strains were estimated by forced overwhelming pneumococcal infection challenge, giving induction values above those obtained for the Cd injections in the Balb/C strain and equal to those obtained for Cd in the more responsive C3H/HeJ strain. Together with direct defendants against harmful oxygen radicals, MT may act complementary as a radical scavenger and by Cd inactivation, but to a higher degree in the C3H/HeJ compared to the Balb/C strain. PMID- 10840638 TI - The rat visceral yolk sac internalizes maternal transferrin and secretes hydrolyzed products towards the fetus. AB - The uptake of transferrin by the rat visceral yolk sac membranes, and the fate of this protein, were measured in a two-chambered system which allowed access to both surfaces of these membranes, i.e. that facing the maternal compartment and that facing the fetal compartment. 125I-labeled transferrin was internalized by the maternal surface of the visceral yolk sac but not by the fetal surface. Following internalization, this transferrin was degraded and the amino acids were secreted exclusively towards the fetal compartment. Transcytosis of intact transferrin was not detected in either direction. These results suggest that transport across the rat visceral yolk sac bound to maternally derived transferrin is not a major mechanism of iron transport in vivo. These results support a role for the visceral yolk sac in fetal metabolism, or supplying the fetus with amino acids derived from degradation of specific maternal plasma proteins, in this case, transferrin. PMID- 10840639 TI - Structural characterization and transcriptional pattern of two types of carp rhodopsin gene. AB - This work characterizes the genomic structures of two types of carp (Cyprinus carpio) rhodopsin (cRh) gene, i.e. type I (cRh-I) and type II (cRh-II). Two types of cRh gene share only 45.6% polynucleotide identity in the upstream region from nucleotide -3436 to +97. However, three conserved regions are found. Homologies to the consensus recognition sites for transcription factors, Crx and Nrl, which are involved in photoreceptor-specific expression, are also observed in cRh genes. With specific polymerase chain reaction (PCR) primers, the two types of cRh gene can be clearly discriminated from each carp genome. Most carps exhibit both types of cRh gene, however, there are still carps possessing either cRh-I or cRh-II. Both cRh-I and cRh-II mRNAs are expressed at an approximately equal level in both eyes extracted from a carp carrying both types of cRh gene. PMID- 10840640 TI - Foreign gene expression by in vivo gene electroporation in the quail testis. AB - To investigate whether or not foreign gene expression is attained in the testis of living Japanese quails, a firefly luciferase reporter gene was transfected by in vivo electroporation (EP), and transcriptional activity of different promoters was compared. In addition, the effect of the Epstein-Barr virus self-replication sequence was also tested. The results showed that luciferase activity in the testis reached almost a plateau value at 50 V. Under this EP condition, no difference was found in transcriptional activity between the simian virus 40 (SV40) and miw promoters. The reporter gene expression in the quail testis was observed over 28 days after in vivo gene EP, although the activity gradually decreased, and the presence of the Epstein-Barr virus self-replication sequence in the SV40 promoter did not significantly prolong the luciferase activity. These results suggest that in vivo gene EP confers strong, though transient, foreign gene expression in the Japanese quail, and it may provide a new powerful approach for studies on transcriptional regulation of genes during proliferation and differentiation of spermatogenic cells in the quail testis. PMID- 10840641 TI - Synthesis and organization of vitellogenin and vitellin molecules from the land crab Potamon potamios. AB - We have previously reported that vitellogenin (Vg) of some female animals contained four polypeptides with molecular mass of 181, 115, 105 and 85 kDa, whereas Vg of most animals contained three polypeptides with molecular mass of 115, 105 and 85 kDa. In the present investigation, we examined whether the 181 kDa polypeptide is the precursor of 115 and 105 kDa Vg and vitellin (Vn) polypeptides. Labeling studies, using [35S]methionine on normal vitellogenic animals, showed that the radioactivity was distributed first among the 181 and 85 kDa polypeptides. SDS-PAGE analysis of purified hemolymph Vg from eyestalk ablated female animals revealed in most animals two polypeptides with an apparent molecular mass of 181 and 85 kDa. These results from in vivo experiments corroborated the view that the 115 and 105 kDa Vg and Vn polypeptides are derived from heaviest 181 kDa polypeptide. In addition it was demonstrated that hepatopancreas and ovary of Potamon potamios incubated in vitro with [35S]methionine synthesized five polypeptides with apparent molecular mass of 224, 181, 115, 105, and 85 kDa while the hepatopancreas appeared to secrete the 181, 115, 105 and 85 kDa polypeptides. The major 115, 105 and 85 kDa polypeptides were found to be components of egg Vn, while the 224 kDa polypeptide was found to be minor component of Vg and Vn from hepatopancreas and ovary extracts, respectively. We conclude that the Vn polypeptides produced by ovary are similar to those produced by hepatopancreas. PMID- 10840642 TI - beta-Hydroxybutyrate in developing nauplii of brine shrimp (Artemia franciscana K.) under feeding and non-feeding conditions. AB - Body content of beta-hydroxybutyrate, and individual dry mass, carbon content, and survival rate, were studied in developing nauplii of the brine shrimp Artemia franciscana K. from hatching to 96-97 h post hatching at 27 +/- 1 degrees C. The effect of two diets was studied in the experiment: Super Selco (SS) with a high lipid content; and Protein Selco (PS) with a high protein content. A starving group (S) was used as reference. The level of beta-hydroxybutyrate at hatching was 0.6 nmol.ind-1; it increased to 1.0-1.5 nmol.ind-1 in the SS- and S-groups, while in the PS-group it remained stable between 0.6-0.8 nmol.ind-1. At 60-80 h post hatch in the SS- and S-groups, the levels of beta-hydroxybutyrate were similar to the initial levels. The survival rate remained higher than 95% until 24 h post hatching in all groups. At the end of the experiment, the survival rate was 63% in the PS-group, 13% in the S-group and 3% in the SS-group. The Artemia nauplii individual dry mass and carbon content remained relatively stable in the SS-group; both parameters showed a significant increase in the PS-group and a significant decrease in the S-group. The results suggest that Artemia nauplii utilise ketone bodies as a fuel during development and growth, but that ketone catabolism may be overloaded by excessive lipid feeding resulting in increased mortality and possibly a ketotic acidosis. PMID- 10840643 TI - Effects of temperature on the respiration rates and the kinetics of citrate synthase in two species of Idotea (Isopoda, Crustacea). AB - The two species of isopods, Idotea baltica (Pallas) and Idotea emarginata (Fabricius), co-occur frequently near Helgoland, North Sea, occupying different ecological niches. Respiration rates and kinetic properties of citrate synthase (CS) were compared in these species in order to identify possible mechanisms of temperature adaptation. Specimens were acclimated to 5 and 15 degrees C prior to further investigations. Respiration rates were measured under normoxic conditions at 5, 10 and 15 degrees C. CS was partly purified chromatographically and influences of temperature, pH, substrate saturation and ATP-concentration on enzyme activity were examined. In both species, rising temperatures led to linearly increasing oxygen consumption, with estimated Q10 values between 3.2 and 4.2. Only I. baltica showed an effect of short term acclimation: warm adapted animals had always higher respiration rates than cold adapted ones. In I. emarginata, the acclimation temperature had no effect on oxygen consumption. Furthermore, its CS slightly indicates higher affinity to oxaloacetic acid when specimens were adapted to 15 degrees C compared to those maintained at 5 degrees C. Any effect of the experimental temperature on CS in I. baltica was negligible. The results are discussed in view of the different habitats occupied by the species compared. PMID- 10840644 TI - Neutral calcium-activated proteases from European sea bass (Dicentrarchus labrax L.) muscle: polymorphism and biochemical studies. AB - Calcium-dependent proteinases or calpains were studied in fish muscle. Hydrophobic chromatography, followed by anion-exchange chromatography of the soluble fraction of sea bass white muscle proteins, resulted in three peaks of calcium-dependent protease activity at neutral pH (A, B and C). They are all neutral cysteine calcium-activated proteinases and can, therefore, be classified as calpain-like enzymes. From the Ca2+ concentration required for activity, A is a mu-calpain, and B and C are m-calpains. They share many properties with calpains from other vertebrate cells but differ in native mass, subunit composition, and the unusual numbers in which they are present. Their specific pattern of expression throughout the year could be of great importance to the resulting rate and extent of degradation of fish flesh after death. PMID- 10840645 TI - Ribose 1,5-bisphosphate inhibits fructose-1,6-bisphosphatase in rat kidney cortex. AB - Fructose-1,6-bisphosphatase is one of the regulatory enzymes of gluconeogenesis in kidney cortex. The effect of ribose 1,5-bisphosphate on fructose-1,6 bisphosphatase purified from rat kidney cortex was studied. Rat kidney cortex, fructose-1,6-bisphosphatase exhibited hyperbolic kinetics with regard to its substrate, but the activity was inhibited by ribose 1,5-bisphosphate at nanomolar concentrations. The inhibitory effect of ribose 1,5-bisphosphate on the fructose 1,6-bisphosphatase was enhanced in the presence of AMP, one of the inhibitors of fructose-1,6-bisphosphatase. Fructose-2,6-bisphosphate, which is an inhibitor of fructose-1,6-bisphosphatase, inhibited rat kidney cortex fructose-1,6 bisphosphatase activities at a low concentration of fructose-1,6-bisphosphate but a high concentration of fructose-1,6-bisphosphate relieved fructose-1,6 bisphosphatase from fructose-2,6-bisphosphate-dependent inhibition. On the contrary, fructose-1,6-bisphosphate was not effective for the recovery of fructose-1,6-bisphosphatase from ribose 1,5-bisphosphate-dependent inhibition. These results suggest that ribose 1,5-bisphosphate is a potent inhibitor and is involved in the regulation of fructose-1,6-bisphosphatase in rat kidney cortex. PMID- 10840646 TI - The effects of hydrostatic pressure on pertussis toxin-catalyzed ribosylation of G proteins from deep-living macrourid fishes. AB - To test the effects of hydrostatic pressure on the coupling of receptors to guanyl nucleotide binding reglatory proteins (G proteins) in transmembrane signaling, pertussis toxin (PTX)-catalyzed [32P]ADP-ribosylation was used to probe the guanyl nucleotide-binding proteins Gi and G(o) in brain membranes from four marine teleosts. These macrourids, Coryphaenoides pectoralis, Coryphaenoides cinereus, Coryphaenoides filifer and Coryphaenoides armatus, span depths from 200 to 5400 m. Pertussis toxin specifically labelled proteins of 39-41 kDa. The PTX catalyzed [32P]ADP-ribosylation reaction was linear for 7 h. Added guanyl nucleotides (guanosine 5'-diphosphate (GDP) and guanosine 5'-O-(3 thiotriphosphate)(GTP[S])) at concentrations up to 1000 microM did not affect ribosylation at atmospheric pressure. Under basal conditions the Gi/G(o) protein population appears to be uncoupled from receptors and bound with GDP. Pressures up to 476 atm were tested in the absence and presence of added guanyl nucleotides, 100 microM GDP and 100 microM GTP[S]. [32P]ADP-ribosylation in brain membranes from the deeper-occurring C. cinereus, C. filifer and C. armatus was not inhibited by increased pressure in the presence of 100 microM GDP. Increasing pressure decreased ribosylation in brain membranes of C. pectoralis. In the presence of 100 microM GTP[S], increased pressure inhibited ribosylation in all species. Pressure appears to enhance the efficacy of GTP[S] in dissociating the heterotrimeric holoprotein. PMID- 10840647 TI - Allozymic variation of Hynobius kimurae Dunn (Amphibia, Caudata). AB - An electrophoretic survey was conducted to examine genetic divergence among 21 populations of a lotic-breeding salamander Hynobius kimurae from Honshu, Japan. Genetically H. kimurae proved to be specifically distinct from H. naevius. Hynobius kimurae is divided genetically into two groups of local populations, and the five populations from the eastern area are genetically distinct from the remaining populations of the central and western areas. Less prominent genetic differentiation was revealed between the western and central populations. Separation of the eastern and central-western groups are discussed in relation to the formation of the Japanese mainland. PMID- 10840648 TI - Molecular analysis of two genes, DD9A and B, which are expressed during the postmolt stage in the decapod crustacean Penaeus japonicus. AB - In decapod crustaceans, deposition of calcium carbonate crystals (calcification) in the exoskeleton takes place during the postmolt phase of the molt cycle. In an attempt to identify proteins which regulate the calcification process, the differential display technique was used to identify genes which were specifically expressed in the integument during the postmolt stage in the penaeid prawn Penaeus japonicus. One of the genes thus identified, named DD9A, was expressed in the epithelial cells of the tail fan. DD9A encoded a putative precursor of a secreted protein of 113 amino acids which exhibited sequence similarities to a group of crustacean and insect cuticular proteins, suggesting that DD9A was a protein component of the exoskeleton. Another gene, DD9B, which was also transcribed specifically during the postmolt period was identified based on its sequence similarity to DD9A. Potential roles of the DD9A protein in the calcification of the exoskeleton will be discussed. PMID- 10840649 TI - Purification and characterization of a fibrinolytic enzyme and identification of fibrinogen clotting enzyme in a marine green alga, Codium divaricatum. AB - A fibrinolytic enzyme was isolated from a marine green alga, Codium divaricatum, and designated C. divaricatum protease (CDP). This protease effectively hydrolyzed fibrinogen A alpha chain, while it had very low hydrolyzing efficiency for B beta and gamma chains. This property was similar to that of alpha fibrinogenase isolated from snake venom. Protease activity peaked at pH 9, and was completely inhibited by diisopropyl fluorophosphate (DFP) and phenylmethylsulfonyl fluoride (PMSF), identifying it as a serine protease. Its molecular form was single polypeptide structure and molecular weight was estimated as 31,000 by SDS-PAGE. Fibrinogen clotting enzyme was also identified in a fraction by ion-exchange chromatography. Analysis of clots formed by the enzyme and by thrombin by SDS-PAGE showed that the fibrinogen clotting enzyme would act like thrombin and have high substrate specificity. PMID- 10840650 TI - Reliability and validity of the Psychosocial Risk Factors Questionnaire (PRFQ). AB - The Psychosocial Risk Factors Questionnaire (PRFQ) was developed to measure perceived attractiveness, concern about physical appearance, and social pressure for thinness. These variables have been identified as correlates of eating disorder symptoms. The PRFQ has four subscales: Social Pressure for Thinness, Media Pressure for Thinness, Concern for Physical Appearance, and Perception of Physical Appearance. This study assessed its reliability and validity. Seventy two women enrolled in undergraduate psychology courses completed the PRFQ and measures selected to assess the concurrent validity of its four subscales. Its test-retest reliability was tested in a subsample of 60 women. Support was found for the test-retest reliability, internal consistency and construct validity of all four subscales. PMID- 10840651 TI - Global negative self-evaluations, weight and eating concerns and depressive symptoms: a prospective study of adolescents. AB - A longitudinal study with four sequential cohorts of girls and boys in early to mid-adolescence (n = 607) was used to assess adolescent changes in global negative self-evaluation, depressive symptoms and weight and eating concerns. The effects of these areas on one another over time were also assessed. Over the six months between the two data collection times, both the girls and the boys experienced an improvement in depressive symptoms, while the boys also experienced a decrease in weight concerns. The results indicated that global negative self-evaluations could predict weight and eating concerns in boys and girls, while weight concerns could predict depressive symptoms in girls. These results are of particular interest, since these relationships have not been previously examined in a longitudinal study. PMID- 10840652 TI - Eating disorders and family therapy. Why, how and when? AB - This article is based on a lecture delivered to the 2nd Nordic Congress on Eating Disorders in Bergen, Norway, 1998, updated to 2000. It examines how earlier ages looked upon and therapeutically evaluated family interaction with regard to the young anorexic patient. The author presents several foreground figures in the field of family therapy thinking, submits his own research in the field, outlines his current therapeutic practice, and concludes with his own answers to the questions: why, how and when? PMID- 10840653 TI - Daily hunger sensation monitoring as a tool for investigating human circadian synchronization. AB - This study investigates within-day hunger sensation (HS) variability in Clinically Healthy Subjects Adapted to Living in Antarctica (CHSALA), as compared to their coeval subjects living in their mother country. The aim is to detect how the orectic stimulus behaves in those environmental conditions and occupational schemes, in order to investigate the individual synchronization to sleep-wake alternation and meal time schedule. HS was estimated via a self-rating score of its intensity on a Visual Analog Scale, repeating the subjective perception every 30 min, unless sleeping. The individual HS time-qualified scores (orexigram) were analyzed according to conventional and chronobiological procedures. The orexigrams of the CHSALA were seen to show a more cadenced intermittence during the diurnal part of the day, strictly related to the meal timing, and a preserved circadian rhythm as well. In addition, these orexigrams were resolved in a spectrum of harmonic components which indicated a subsidiary number of ultradian formants. These findings are convincing evidence that the individual orexigram may be used to investigate whether or not a single subject is synchronized to sleep-wake cycle, meal time schedule and socio-occupational routines, instead of using more complex and expensive techniques, involving automated equipments and biohumoral assays. PMID- 10840654 TI - Eating attitudes and behavior throughout the menstrual cycle in obese women: a case-control study. AB - Premenstrual variations of eating behavior are reported in several studies, but their relationship with mood is unclear. Eating behavior and physical and psychological complaints during the menstrual cycle were studied in 107 obese patients and 93 matched controls using retrospective (Weekly Bulimic Test Edinburgh, W-BITE and Premenstrual Assessment Form, PAF) and prospective (Daily Rating Form, DR) questionnaires. Eating disorder symptoms increased in the premenstrual phase, as shown by the W-BITE scores both in patients (6.2 +/- 5.3 premenstrual week vs 4.9 +/- 4.4 postmenstrual week, p < 0.05) and in control subjects (4.9 +/- 4.1 premenstrual week vs 4.2 +/- 3.0 postmenstrual week, p < 0.05) and were correlated to premenstrual complaints in control subjects (r = 0.5; p < 0.05) but not in obese women (r = 0.2; p = NS). A close relationship between physical and psychological premenstrual disturbances was observed in obese patients only. Premenstrual variation of eating behavior could be the target of specific treatment. PMID- 10840655 TI - Body image and eating restraint: a structural modeling analysis. AB - Body-image disturbance and its link with eating behaviors are poorly understood and controversial phenomena. The model proposed by Cash (1996) is perhaps the most unified current attempt to analyze this link. Its central features are a separation of historical and proximal influences and the link between body image emotions and adjustive, emotion regulating actions. This model, however, and in particular the link between body dissatisfaction and eating restraint have not been empirically tested. A series of different causal models for directly observed variables (LISREL 7) have therefore been used to determine the soundness of this link in a study of 130 female undergraduates. The goodness-of-fit of the models suggests that a causal link exists. It is clear that body image therapy should be combined with current treatments for obesity and eating disorders. PMID- 10840656 TI - Depressive symptoms among obese children. AB - The authors compare the appearance of depressive symptoms in 30 obese children in outpatient care and 30 normal-weight controls. The Montgomery-Asberg Depression Rating Scale showed a significantly higher rate of depression in the obese children (p < 0.01). The results are interpreted in the light of the partially contradictory views in the literature. Reference is also made to the therapeutic aspects of the problem. PMID- 10840657 TI - Factor structure of the Eating Attitudes Test (EAT) in a Turkish university sample. AB - OBJECTIVE: The primary purpose of this study was to compare the factor structure of the Eating Attitudes Test (EAT) in a sample of Turkish university students with both that of the original Canadian study and that of a Turkish study conducted ten years earlier. METHOD: The EAT 40 was administered to a sample of 270 female undergraduates at the Bogazici and Marmara universities in Istanbul, Turkey. RESULTS: The factor structure produced more closely resembled the original than did that of the study conducted ten years ago. Of particular note was the presence of a bulimia factor which had not emerged in the previous Turkish study, together with a new factor labelled "Ambivalence about Eating". DISCUSSION: Comparison of the two Turkish studies highlights the emergence of a bulimia factor not present ten years ago. This reflects clinical experience. Moreover the new Ambivalence about Eating factor is discussed in terms of the transitional nature of Turkish society and the role of a culture clash between traditional and westernized attitudes towards food. It is therefore predicted that the prevalence of eating disorders in Turkish culture will dramatically increase in the foreseeable future. PMID- 10840658 TI - Screening for sickle cell disease and thalassaemia: a systematic review with supplementary research. PMID- 10840659 TI - [Drug therapy of heart failure]. PMID- 10840660 TI - [Drug therapy of essential hypertension]. PMID- 10840661 TI - [Drug therapy of cardiac arrhythmias]. PMID- 10840662 TI - [Inhibition of thrombocyte aggregation as a therapeutic principle. Review of the literature and currently approved thrombocyte aggregation inhibitors]. PMID- 10840663 TI - [Stroke prevention with thrombocyte function inhibitors and anticoagulants]. PMID- 10840664 TI - Reducing adverse drug events: lessons from a breakthrough series collaborative. AB - BACKGROUND: In January 1996, 38 hospitals and health care organizations (for a total of 40 hospitals) in the United States came together in an Institute for Healthcare Improvement (IHI; Boston) Breakthrough Series collaborative to reduce adverse drug events-injuries related to the use or nonuse of medications. METHODS: The participants were taught the Model for Improvement, a method for rapid-cycle change and evaluation, and were then coached on how to identify their own problem areas and develop changes in practice for rapid-cycle testing. These changes could be implementation of one or more known medication error prevention practices or new practices developed. RESULTS: During a 15-month period the 40 hospitals conducted a total of 739 tests of changes. Process changes accounted for 63% of the cycles; the remainder consisted of preliminary data gathering, consensus-building, or education cycles. Eight types of changes were implemented by seven or more hospitals, with a success rate of 70%. These changes included non-punitive reporting, ensuring documentation of allergy information, standardizing medication administration times, and implementing chemotherapy protocols. DISCUSSION: Success in making significant changes was associated with strong leadership, effective processes, and appropriate choice of intervention. Successful teams were able to define, clearly state, and relentlessly pursue their aims, and then chose practical interventions and moved early into changing a process. They did not spend months collecting data before beginning a change. Changes that were most successful were those that attempted to change processes, not people. Health care organizations committed to patient safety need not regard current performance limits as inevitable. PMID- 10840665 TI - Reducing medication errors in hospitals: a peer review organization collaboration. AB - BACKGROUND: Motivated by published reports of the incidence, costs, causes, and nature of adverse drug events (ADEs) in hospitalized patients, in 1997 the Medicare peer review organization for Nevada and Utah initiated a voluntary project of medication error reduction for Utah hospitals. METHODS: Through project activities, hospital teams were encouraged to make changes to their medication processes based on direct evaluation of medication systems characteristics, informed by ergonomic principles and published studies of medication errors. Assessment of project effects included an evaluation of the changes implemented and results from an anonymous medication errors survey of clinical staff from participating organizations. RESULTS: Thirteen of the 39 acute care hospitals in Utah participated in 1997-1998 in the collaborative project. Participants reported substantive medication system changes that were expected to result in improved patient safety. Baseline and follow-up survey data were available for 8 of the participating hospitals. Analysis of 560 responses showed a 26.9% decrease in overall error frequency, a 12.5% increase in error detection and prevention, and a 24.1% increase in formal written reporting of errors that reached the patient. CONCLUSIONS: This project demonstrated community interest in a proactive and collaborative approach to improving patient safety. The improvement efforts were substantive and sustainable. Survey results suggest that the changes implemented in participating organizations may have reduced medication errors and improved capacity for error detection and prevention. PMID- 10840666 TI - Making medication safety a strategic organizational priority. AB - Since January 1998 the Executive Session on Medical Error and Patient Safety at Harvard's John F. Kennedy School of Government has periodically convened a group of 25 to 30 practitioners (and a few academics) to discuss issues and identify strategies and solutions concerning patient safety. This profile is adapted from a case study presented at the Executive Session. PMID- 10840667 TI - Patient expectations for surgery: are they being met? AB - BACKGROUND: The purpose of the study was to determine patient expectations for the outcomes of three elective surgical procedures, the extent to which patient expectations for surgery were met, the reasons for unmet expectations, and the factors that might predict unmet expectations. Better understanding of these questions might help identify targeted interventions to better prepare patients for specific health care experiences. METHODS: In a longitudinal, prospective design, a convenience sample of 445 patients (age range, 18 to 86 years) at a general surgery clinic at a major academic medical center was included--177 patients undergoing inguinal hernia repair, 146 undergoing parathyroidectomy, and 122 undergoing cholecystectomy. Patients completed both standardized and newly developed condition-specific health survey instruments. Preoperative interviews were administered, followed by mailed surveys 2 months after surgery. RESULTS: Between 9% and 27% of the respondents reported unmet expectations, with significant variation by condition; reasons included perceived lack of symptom relief, surgical complications, and process of care issues. Patients undergoing parathyroidectomy had a greater probability of unmet expectations. Both feeling prepared for surgery and improved postoperative symptom relief and role functioning reduced the probability of unmet expectations. DISCUSSION: To reduce the level of unmet expectations, patients need to be prepared both for the surgical experience and for what to expect in the recovery phase. This is especially true for complex illnesses such as primary hyperparathyroidism. Innovative educational strategies to ensure adequate preparation for surgery will be needed, and attention will need to be paid to latent, unstated process measures, if unmet expectations are to be reduced. PMID- 10840668 TI - Computerizing incident reporting at a community hospital. AB - BACKGROUND: A community hospital in Seattle, Northwest Hospital (NWH), reengineered its paper-based incident reporting system into a computerized reporting, notification, and tracking tool. In July 1996 a small interdisciplinary working group was formed to improve the incident reporting system so that it would decrease the time needed to complete an incident report, collect more precise data about the incident, allow department managers instant access to all open memos involving or generated by their departments, allow ad hoc reporting by managers and administration, and allow only involved parties access to memos. IMPLEMENTATION AND EVALUATION: After a pilot study was conducted in the Childbirth Center and the pharmacy department began using the computerized incident reporting system, other units began using the system according to a weekly roll-out schedule during the first two quarters of 1998. In the third quarter of 1998, NWH began using the system as its primary quality assurance and incident reporting tool. As soon as an incident is documented, it is in the database and available for reporting. Data collected from January 1998 through December 1999 indicated that turnaround time for the life cycle of an incident report decreased from 53 to 12 days. At least 20 hours a month were saved in transcription and data entry time using the new system. DISCUSSION: Although incident reporting is now more streamlined and efficient, a few issues have emerged that need to be addressed, some relating to users' incorrect entering of information. Improvements are still being made to the system on an ongoing basis. PMID- 10840669 TI - Effects of supplementation of dairy cattle with fish oil on silage intake, milk yield and milk composition. AB - The effects of level of fish oil inclusion in the diet on grass silage intake, and milk yield and composition of dairy cows offered either 5 or 10 kg concentrates/d were evaluated in a ten treatment, partly balanced, changeover design experiment involving 50 cows in early lactation. Concentrates were prepared to provide 0, 150, 300 or 450 g fish oil/cow per d or 300 g fish oil/cow per d from a premix when each animal was offered 5 kg/d. The fish oil was predominantly from herring and mackerel caught in the North Atlantic while the fish oil premix was obtained from a commercial source and used palm kernel expeller as a carrier. Increasing fish oil supplementation decreased silage dry matter intake and the concentrations of milk fat and protein, and increased milk yield and diet digestibility. There were significant interactions between concentrate feed level and level of fish oil for silage intake and milk yield. Other than for the concentrations of milk fat and protein, and 20:4n-6 fatty acids, the source of fish oil did not affect forage intake or animal performance. Fish oil supplementation also decreased the concentrations of milk protein by 0.9 g/kg for each 100 g increase in fish oil supplementation, the depression being similar at each level of concentrate feeding. Supplementing the feed of dairy cows with 450 g fish oil/cow per d decreased the concentration of milk fat by 15 g/kg. This study also showed that feeding dairy cattle with fish oil is an efficient method of increasing eicosapentaenoic acid in the human diet through transfer into milk. PMID- 10840670 TI - Mammary infection with Staphylococcus aureus in cows: progress from inoculation to chronic infection and its detection. AB - The progress of Staphylococcus aureus infection from inoculation to the early chronic stage was examined in 12 Israeli-Holstein cows (four primiparous and eight multiparous) for up to 48 d after inoculation. Before inoculation, the primiparous cows were free from any infection and the multiparous cows were infected by coagulase-negative staphylococci. Two quarters in each cow were inoculated intracisternally following milking with 2000 cfu of a local prevailing Staph. aureus strain, VL-8407. Infection was established in 21 out of 24 quarters. The control quarters remained free from infection during the study, with no significant change in function. No statistically significant differences were found between primiparous and multiparous cows in the responses examined. Somatic cell count (SCC) increased within 24 h of inoculation and remained high for the duration of the study. In the infected quarters mean ln (SCC) increased within 24 h from 9.9 +/- 0.5 before inoculation to 13.0 +/- 0.2 after inoculation; most of the cells were neutrophils. N-acetyl-beta-glucosaminidase activity, expressed as ln (nnmol/min per l), was increased from 0.9 +/- 0.6 to 2.4 +/- 0.2 by inoculation, and was highly correlated with SCC. The Staph. aureus count fluctuated with no particular relationship with SCC. The phagocytic activity of neutrophils was significantly lower in the inoculated than in the control quarters and this difference increased with time after inoculation. CD8+ T lymphocytes were the main subpopulation of lymphocytes found in inoculated quarters. After inoculation, maximum but not minimum electrical conductivity (EC) recorded during milking increased significantly. The rises in maximum EC varied significantly among cows. The rises in SCC were associated with a persistent increase in EC in only one of the eight cows examined. No clinical signs were observed, and milk yield and composition were not affected during the study period. The results suggest that some strains of Staph. aureus may induce a relatively mild response in mammary glands of cows in mid lactation, and that the concomitant development of such chronic Staph. aureus infections in two quarters may not be detected by changes in the EC of composite milk and in the yield of the cow. PMID- 10840671 TI - Simulation analysis of substrate utilization in the mammary gland of lactating cows. AB - A kinetic modelling approach was developed and investigated with the aim of predicting the utilization of major substrates in the mammary gland and milk secretion rates in the lactating cow at varying concentrations of substrate in arterial blood. The model includes kinetic equations of transport and metabolism of glucose, acetate, free amino acids and free fatty acids in secretory cells and a phenomenological description of autoregulation of local blood flow, in which an energy criterion of control has been used. The predicted relationships between the rate of milk secretion and glucose levels in the blood are consistent with experimental results. Differential stimulation of alpha-lactalbumin synthesis causes increments in local blood flow and milk secretion rate in the model. The results of the study suggest that there is no simple relationship between the level of substrates in the blood and milk yield and contents of fat and protein in milk. This is because the effect on production of varying patterns of substrate concentrations in the blood is mediated by network interactions at the level of secretory cell metabolism and microcirculation. However, dynamic modelling provides a rational framework for developing such predictive tools. PMID- 10840672 TI - Effects of yeast expressed recombinant interleukin-2 and interferon-gamma on physiological changes in bovine mammary glands and on bactericidal activity of neutrophils. AB - The physiological effects of intramammary infusions of recombinant bovine cytokines in six lactating dairy cows on the quality and yield of milk and the bactericidal activity of milk neutrophils were investigated. Recombinant bovine interleukin-2 (rboIL-2) and interferon-gamma (rboIFN-gamma) were produced in the yeast Pichia pastoris. Two animals were given rboIL-2 (2 x 10(5) units) in two quarters, two animals were given rboIFN-gamma (6.5 x 10(5) units) in two quarters, and the other two cows received a dose of rboIL-2 in one quarter and rboIFN-gamma in a second quarter. In addition, each animal was given phosphate buffered saline (PBS) in the other two quarters as a control. Somatic cell counts and conductivity of the fore milk were monitored before and after infusion. Neutrophils were isolated from quarter milk samples 36 h after infusion of cytokine or PBS and their bactericidal activities against Staphylococcus aureus were measured in vitro with a colorimetric assay. Quarters infused with rboIL-2 or rboIFN-gamma showed significant but transitory increases in both milk somatic cell counts and conductivity when compared with preinfusion values and with control quarters. There were minimal effects on daily milk yield. Neutrophils isolated from milk from quarters infused with rboIL-2 showed enhanced bactericidal activity against Staph. aureus. The bacterial killing from rboIL-2 treated quarters was significantly greater, with a mean of 63.5% compared with a mean of 5.4% for neutrophils taken from uninfected quarters to which PBS had been administered. The bactericidal activities for quarters treated with rboIFN-gamma and infected quarters treated with PBS were 15.0 and 30.0% respectively. The results indicate that intramammary infusions of rboIL-2 and rboIFN-gamma to lactating cows are well tolerated, and that rboIL-2 can activate milk neutrophils and augment their bactericidal activity. PMID- 10840673 TI - Effect of storage at 4 degrees C on the physicochemical and renneting properties of milk: a comparison of caprine, ovine and bovine milks. AB - The effects of cold storage at 4 degrees C for 12, 24 and 48 h on the physicochemical characteristics and renneting properties of ewes', goats' and cows' milks were compared. The most important changes were observed in cows' milk. Soluble calcium concentrations were not affected in ewes' milk but were increased by 10% in cows' milk and 7% in goats' milk. More casein was dissociated on cooling cows' (+300%) than goats' (+100%) milk, and there was no change in soluble casein in ewes' milk. The coagulation characteristics of cows' milk were more impaired by cold storage than goats' or ewes' milks. Coagulation times increased by approximately 30% and whey draining capacity decreased by 40% after cows' milk was cooled, but there were no changes with ewes' milk and only a slight decrease in coagulation time with goats' milk. We propose an interpretation of these results based on the physicochemical properties of each type of milk. PMID- 10840674 TI - Capillary electrophoresis characterization of the casein fraction of cheeses made from cows', ewes' and goats' milks. AB - Casein fractions and their breakdown products in Iberico-type cheeses made from the milk of cows, ewes or goats were analysed by capillary electrophoresis in order to characterize them. The actions of plasmin and chymosin on caseins were evaluated by comparing the electropherograms of caseins from milk and from cheese, both with and without treatment with plasmin. Characteristic capillary electrophoresis patterns were obtained for cheeses made from the milk of each of the three species, and the main components were identified. Caprine para-kappa casein and bovine beta-caseins, eluting at the first and at the last part of the electropherogram respectively, were found to be indicative of the presence of the milks of these species. PMID- 10840675 TI - Characterization of genetic polymorphism in the goat beta-lactoglobulin gene. AB - Two new variants have been detected and characterized for the goat beta lactoglobulin gene at the cDNA level and confirmed at the genomic level. The two polymorphisms are located on exon 7 of the gene. One of the polymorphic sites is produced by a single nucleotide substitution in position +4601, allowing a polymerase chain reaction--restriction fragment length polymorphism (PCR-RFLP) genotyping procedure to be developed using SacII restriction enzyme. The other polymorphic position contains a 10 bp long insertion at position +4641 that can be detected by capillary electrophoresis of the PCR product amplified with a fluorescent primer. The association of these two polymorphisms was also investigated, resulting in the description of two new alleles. Both of these contained the point mutation at the SacII site, with or without the 10 bp insertion at position +4641. The distribution of these new polymorphisms was studied in a population of males of four different goat breeds. The gene frequencies for these variants were similar in Spanish and French breeds. PMID- 10840676 TI - Stability of vitamins during the storage of liquid infant milks. AB - The stability of the water-soluble B vitamins and the fat-soluble vitamins A and E of three liquid infant milks was followed throughout 12 months of storage at 20, 30 and 37 degrees C. Two samples were 'follow-on' milks from two different manufacturers and the other a 'junior' milk from one of these. The levels of water-soluble vitamins and vitamin E remained constant during storage at the three temperatures. In contrast, vitamin A decreased in samples from one manufacturer at all storage temperatures, and in samples from the other only at 37 degrees C. A kinetic equation was derived that made it possible to predict the influence of storage temperature on the shelf life of these kinds of products, together with the storage period during which levels of vitamin A recommended as adequate would remain. PMID- 10840677 TI - Accumulation of casein-derived peptides during growth of proteinase-positive strains of Lactococcus lactis in milk: their contribution to subsequent bacterial growth is impaired by their internal transport. AB - To explain the limited nutritional value of milk cultured with proteinase positive (Prt+) strains of Lactococcus lactis for the subsequent growth of dairy lactococci, we investigated further the time courses of modifications in the free amino acid and peptide contents of cultured milk. When growing in milk for up to 24 h, Prt+ strains of Lc. lactis progressively accumulated amino acids and casein derived peptides. The growth of proteinase-negative (Prt-) wild-type strains and peptide transport mutants of Lc. lactis in cultured milk showed that casein derived peptides could sustain growth up to 5 x 10(8) cfu/ml, depending on the extent of casein degradation during the preliminary growth of Prt+ strains and the Prt- strains. Of the casein-derived oligopeptides, < 25% were transported into the cell and used for Lc. lactis growth. However, they played a prominent role, contributing 90% to growth. In contrast, di- and tripeptides did not contribute to growth, suggesting that either few were released from caseins or they did not supply essential amino acids. PMID- 10840678 TI - Fatty acid composition and freeze-thaw resistance in lactobacilli. AB - The fatty acid composition and freeze-thaw resistance of eight strains of thermophilic lactobacilli were studied. Seven of these contained the same polar and neutral lipids, the five major components making up 90% of the cellular fatty acid pool being 14:0, 16:0, 16:1, 18:1 and C19 cyclopropane (cyc19:0). Strain comparison by means of cluster analysis based on the fatty acid ratios using the overlap coefficient revealed two well defined clusters. One was formed by three strains of species Lactobacillus delbrueckii subsp. lactis and Lb. delbrueckii subsp. delbrueckii, the other included five strains of the species Lb. delbrueckii subsp. bulgaricus, Lb. acidophilus and Lb. helveticus. Resistance of strains with a high content of unsaturated fatty acids (66-70%) decreased with increasing cyc19:0 concentrations. In contrast, in strains with a low concentration of unsaturated fatty acids (42-49%), increasing cyc19:0 levels were associated with increased freeze-thaw resistance. PMID- 10840679 TI - Chemiluminescence of bovine polymorphonuclear leucocytes during the periparturient period and relation with metabolic markers and bovine pregnancy associated glycoprotein. AB - The respiratory burst activity of polymorphonuclear leucocytes (PMN) was evaluated in eight Holstein cows from 8 weeks before until 6 weeks after calving by chemiluminescence (CL). The CL response started to decrease 1 week before parturition, reaching a minimum during the first 2 weeks after calving. From week 3 of lactation, CL increased again and returned to original levels by week 6 of lactation. Plasma concentrations of 3-hydroxybutyric acid, total bilirubin and bovine pregnancy-associated glycoprotein started to increase before parturition to reach a maximum during the first or second week of lactation. The concentrations of glutamic-oxaloacetic transaminase, lactate dehydrogenase, non esterified fatty acids and bilirubin increased after calving, reaching a maximum during the second week. A small decrease in plasma cholesterol during the week before and after calving was followed by an increase. The CL response of the PMN showed significant temporal relationships with the plasma concentrations of 3 hydroxybutyric acid, bovine pregnancy-associated glycoprotein, bilirubin, glutamic-oxaloacetic transaminase, non-esterified fatty acids; that with cholesterol was nearly significant. This means that the change in the CL response with time coincided with the changes in plasma concentrations of these substances with time and that these changes were significantly related with each other. The results of this study show that the decreased respiratory burst activity of bovine PMN around parturition may be related to the extent of the metabolic and hormonal changes. Although the causative relationships are not proven, these results support earlier results suggesting that 3-hydroxybutyric acid and bovine pregnancy-associated glycoprotein may directly affect neutrophil function, whereas non-esterified fatty acids, cholesterol, bilirubin, and liver enzymes may have potential as diagnostic markers of impaired neutrophil function and consequently increased disease risk around parturition. PMID- 10840680 TI - Autolysis and related proteolysis in Swiss cheese for two Lactobacillus helveticus strains. AB - Intracellular peptidases of Lactobacillus helveticus may play a major role in the proteolysis of Swiss cheeses, provided that they are released through bacterial lysis. Experimental Swiss cheeses were manufactured on a small scale from thermized and microfiltered milk using as starters (in addition to Streptococcus thermophilus and Propionibacterium freudenreichii) one of two Lb. helveticus strains, ITGLH1 and ITGLH77, which undergo lysis to different extents in vitro. All the cheeses were biochemically identical after pressing. The viability of Lb. helveticus ITGLH1 and ITGLH77 decreased to a similar extent (96-98%) while in the cold room, but the concomitant release of intracellular lactate dehydrogenase in cheeses made with strain ITGLH1 was 5-7-fold that in cheeses made with ITGLH77. Protein profiles and immunoblot detection of the dipeptidase PepD confirmed a greater degree of lysis of the ITGLH1 strain. Free active peptidases were detected in aqueous extracts of cheese for both strains, and proteolysis occurred principally in the warm room. Reversed-phase HPLC revealed a more extensive peptide hydrolysis for ITGLH1, which was confirmed by the greater release of free NH2 groups (+33%) and free amino acids (+75%) compared with ITGLH77. As the intracellular peptidase activities of ITGLH1 and ITGLH77 have previously been shown to be similar, our results indicated that the extent of lysis of Lb. helveticus could have a direct impact on the degree of proteolysis in Swiss cheeses. PMID- 10840681 TI - Odour-impact compounds of Gorgonzola cheese. AB - Volatile concentrates were obtained by vacuum distillation from both natural and creamy Gorgonzola cheese and isolated by continuous liquid-liquid extraction. Both were analysed by high resolution gas chromatography (HRGC), HRGC-mass spectrometry and HRGC-olfactometry. A total of 63 components were identified in the neutral extract of the natural type (21 esters, 13 ketones, 14 alcohols, 5 aldehydes, 1 sulphur compound, 7 aromatic compounds and 2 terpenes) and 52 in the creamy type (17 esters, 12 ketones, 10 alcohols, 5 aldehydes, 1 sulphur compound, 5 aromatic compounds and 2 terpenes). Ketones, whose major components were 2 nonanone and 2-heptanone, were the predominant constituents of the neutral fraction. By olfactometric analysis of the neutral extracts, 23 odour-impact compounds were found in the natural and 21 in the creamy Gorgonzola cheese. 1 Octen-3-ol, ethyl hexanoate, 2-nonanone, 2-heptanone, 2-heptanol, ethyl butanoate, 2-nonanol and 4-methylanisole were the key odorants of the natural cheese, whereas 2-heptanone, 2-heptanol, ethyl butanoate, 3-methyl thiopropanal and an unidentified constituent with a fruity odour were characteristic of the creamy Gorgonzola cheese. On the basis of high odour unity values, 2-nonanone, 1 octen-3-ol, 2-heptanol, ethyl hexanoate, methylanisole and 2-heptanone were the most important odorants of natural and creamy Gorgonzola cheese aroma. PMID- 10840682 TI - Milk whey protein concentration and mRNA associated with beta-lactoglobulin phenotype. PMID- 10840683 TI - kappa-casein and beta-Casein alleles in crossbred and Zebu cattle from India using polymerase chain reaction and sequence-specific oligonucleotide probes. PMID- 10840684 TI - Effect of somatic cell count and polymorphonuclear leucocyte content of milk on composition and proteolysis during ripening of Swiss-type cheese. PMID- 10840685 TI - Identification, display, and use of symmetry elements in atomic and electronic structure models. AB - Crystallographic symmetry plays an important role in structure determination from diffraction or scattering data, in spectroscopy and in simulations. It is convenient and insightful to integrate the display and use of such symmetry data with data analysis and modeling methods. We outline the integration of a suite of crystallographic algorithms, closely coupled with interactive graphical displays. These include techniques for identifying the unit cell of a solid, for automatically determining space and point group symmetries, for generalized displays of symmetry elements overlaid on structural models, and for construction, editing, and transformation of models subject to symmetry constraints. In addition, electron densities derived from periodic density functional calculations can be symmetrized and displayed with the corresponding symmetry elements. Applications of these various capabilities in crystallographic research are illustrated by topical examples. PMID- 10840686 TI - Toward minimalistic modeling of oral drug absorption. AB - Poor intestinal permeability of drugs constitutes a major bottleneck in the successful development of candidate drugs. Fast computational tools to help in designing compounds with increased probability of oral absorption are required, since both medicinal and combinatorial chemists are under pressure to consider increasing numbers of virtual and existing compounds. The QSAR paradigm for drug absorption is expressed as a function of molecular size, hydrogen-bonding capacity, and lipophilicity. A nonlinear PLS model that can be achieved with minimal computational efforts is described. The QSAR model correlates human intestinal absorption (%HIA) data, and apparent Caco-2 cell permeability data, to parameters calculated from molecular structures. Two properties were found to be relevant for absorption predictions, namely H-bonding capacity, and hydrophobic transferability. The parsimony principle was applied in several aspects: single conformers were used to compute molecular surface areas; the definitions of "polar" and "nonpolar" surfaces were done in a simplistic fashion; simple and fast 2D descriptors were used to estimate other properties; the 1 PLS component model was selected. These choices result in a minimalistic model for oral absorption. The use of both %HIA and Caco-2 permeability data was found to stabilize and improve the model. This QSAR model can serve as a simple, quantitative extension of the "rule of five" scheme (Lipinski, C.A., Lombardo, F., Dominy, B.W., and Feeney, P.J. Adv. Drug Deliv. Rev. 1997, 23, 3-25), in a manner that can prove beneficial to the drug discovery process. PMID- 10840687 TI - Modeling glasses using the reverse Monte Carlo algorithm: addition of nuclear magnetic resonance and expanded coordination number constraints. AB - In simple oxide glasses the coordination number and oxidation state of the glass forming element can be predicted directly from the "8--n" rule. Tellurite glasses, however, are unusual in that the coordination number of oxygen around tellurium varies without a corresponding change in the oxidation state of tellurium. To model sodium tellurite glasses successfully using the reverse Monte Carlo algorithm several new constraints have been added. Changes include extending the original coordination constraint to allow multiple coordination numbers, and the addition of a new coordination constraint to keep the oxidation state of tellurium constant by limiting the number of bridging and nonbridging oxygens bonded to each tellurium atom. In addition, the second moment of the distribution of dipolar couplings for sodium atoms obtained from a spin-echo NMR experiment was added as a new constraint. The resulting real-space models are presented and the effectiveness of the new constraints is discussed. PMID- 10840688 TI - The structure of electronic states in amorphous silicon. AB - We illustrate the structure and dynamics of electron states in amorphous Si. The nature of the states near the gap at zero temperature is discussed and especially the way the structure of the states changes for energies ranging from midgap into either band tail (Anderson transition). We then study the effect of lattice vibrations on the eigenstates, and find that electronic states near the optical gap can be strongly influenced by thermal modulation of the atomic positions. Finally, we show the structure of generalized Wannier functions for amorphous Si, which are of particular interest for efficient ab initio calculation of electronic properties and forces for first principles dynamic simulation. PMID- 10840689 TI - Peptides quantitative structure-function relationships: an automated mutation strategy to design peptides and pseudopeptides from substitution matrices. AB - The process by which analogs in peptide chemistry are currently designed does not include any quantitative basis for amino acid substitutions from pharmacological leads. Here, we show that substitution matrices such as PAM 250 can provide quantitative constraints compatible with biological activity. This article describes its use in a strategy of rational amino acid substitution in peptides and proteins: we have computed a chemically derived matrix equivalent to the well known PAM 250 matrix, reflecting the natural mutability rates of amino acids in protein evolutions but that can be extended to all the noncoded amino acids. Some of these noncoded amino acids are widely used to mimic secondary structure, to constrain backbone conformation, or to evade protease degradation. An automated sequence mutation (ASM) strategy has been defined to generate mutations within constraints. Application of such a substitution matrix to quantitative structure function relationship studies will be of use in the design of proteins and peptides destined to become pharmaceutical drugs. In particular, issues such as which functionally conserved substitutions are able to satisfy conformational restrictions, oral bioavailability, or formulation demands can be quantitatively addressed. PMID- 10840690 TI - Full window stereo. AB - Visualisation is the bioinformaticist's most important tool for the study of macromolecules, and being able to see molecules in stereo is a crucial aspect. Stereo vision is based on the principle that each eye is presented with the best possible image of what it would have seen if the object was really there in 3D. The simplest approach to stereo vision is to display the right eye picture on the right half of the screen and the left eye picture on the left half while using a mirror system to ensure that each eye sees what it is supposed to see. More expensive workstations use hardware to alternately display the left and right eye pictures while synchronously blocking the transparency in the right or left lens of the special glasses worn by the user. We present here some simple software that uses inexpensive hardware, originally designed for the computer game industry, to make full screen stereo available on Linux-based PCs. The quality of the stereo vision is similar to the top-of-the-line graphics workstations that are capable of quad-buffering. This stereo option has been incorporated in the XII based version of WHAT IF (Vriend, G. J. Mol. Graphics 1990, 8, 52-56), but the stereo source code is freely available and can easily be incorporated in other visualization packages. PMID- 10840692 TI - Drosophila as a genetic tool to define vertebrate pathway players. PMID- 10840691 TI - Improved AMBER torsional parameters for the N-N rotational barrier in diacylhydrazines. AB - The structure and rotational barrier for substituted diacylhydrazines are of significant interest given the role this functionality plays in peptidomimetics and ecdysone agonists, the latter of which have application as extremely selective insecticides. Ab initio calculations show that the lowest energy conformations are typically nonplanar with essentially perpendicular nitrogen lone pairs. Molecular mechanics calculations using the AMBER force field in MacroModel yield minima and rotational barriers that are both quantitatively and qualitatively inconsistent with the ab initio results. In this work the AMBER N-N rotational barriers for all configurations of the parent, methyl and di-methyl substituted diformylhydrazines have been fitted to MP2/6-31 + G relative energies. The resulting AMBER torsional parameters have been validated by calculating the rotational barriers for N-t-butyl substituted diformylhydrazine, dibenzoylhydrazine and an azadipeptide. In each case the new AMBER rotational barriers compare favorably with the corresponding MP2 calculated rotational barriers. PMID- 10840693 TI - Bioassays for studying the role of the peptide growth factor activin in early amphibian embryogenesis. PMID- 10840694 TI - Analysis of mammary gland morphogenesis. PMID- 10840695 TI - Specification of cardiac mesenchyme and heart morphogenesis in vitro. PMID- 10840696 TI - Craniofacial development and patterning. PMID- 10840698 TI - Skeletal morphogenesis. PMID- 10840697 TI - Craniofacial skeletal morphogenesis in vitro. PMID- 10840699 TI - Transplantation and culture techniques for the analysis of urodele limb regeneration. PMID- 10840700 TI - Retroviral infection of T-cell precursors in thymic organ culture. PMID- 10840701 TI - Assay for the isolation of hepatogenic factors. Key molecules in hepatocyte formation and liver morphogenesis. PMID- 10840702 TI - Skin morphogenesis. Embryonic chicken skin explant cultures. PMID- 10840703 TI - Apoptosis in development. PMID- 10840704 TI - Methods to detect patterns of cell death in Drosophila. PMID- 10840705 TI - Mechanisms of neurulation. PMID- 10840706 TI - Neurulation and neural tube closure defects. PMID- 10840707 TI - Neural tube defects. PMID- 10840708 TI - Experimental manipulation and morphometric analysis of neural tube development. PMID- 10840709 TI - Isolation of neuroepithelium and formation of minispheres. PMID- 10840710 TI - Examination of normal and abnormal placentation in the mouse. PMID- 10840711 TI - Palatal dysmorphogenesis. Palate organ culture. PMID- 10840712 TI - Palatal dysmorphogenesis. Quantitative RT-PCR. PMID- 10840714 TI - Assessment of laterality defects in rodent embryos. PMID- 10840713 TI - Transspecies grafting as a tool to understand the basis of murine developmental limb abnormalities. PMID- 10840715 TI - Cardiac morphogenesis and dysmorphogenesis. I. Normal development. PMID- 10840716 TI - Analysis of two aspects of left-right patterning of the vertebrate heart. Heart tube position and heart tube chirality. PMID- 10840717 TI - Biologically based risk assessment models for developmental toxicity. AB - It is obvious that the task of incorporating mechanistic information into dose response assessment for developmental toxicity is, by and large, still at its conceptual stage. Our immature understanding of embryogenesis and teratogenesis forces us to make simplifying biological assumptions that may turn out to be erroneous; therefore, these mechanistically based models should be constructed so as to be easily modified as new information becomes available. The data-intensive (and costly) nature of these modeling efforts may also limit their practice to extraordinary situations where, for instance, large segments of the human population are exposed to low levels of a compound and the determination of a safe level of exposure is of utmost importance, or where compounds are of such immense economic or therapeutic value that their use would warrant a concerted effort to minimize the uncertainties inherent in the current methods of extrapolation. Nevertheless, this chapter has presented several attempts to translate the concept of BBDR into practice. When these applications become successful, these models should provide risk assessors with more reliable response indicators at low doses (where human exposures are realistic) and more accurate cross-strain/cross-species comparisons as well as extrapolations across exposure conditions. Although the BBDR models do not necessarily redefine the current practice of risk assessment using either the NOAEL or the benchmark approach, information derived from these efforts should provide a credible scientific basis for the estimates of RfD. PMID- 10840718 TI - Positional cloning. PMID- 10840719 TI - Gene trapping in embryonic stem cells in vitro to identify novel developmentally regulated genes in the mouse. PMID- 10840720 TI - PCR-based cloning of cortically localized RNAs from Xenopus oocytes. PMID- 10840721 TI - Analysis of mRNA expression during preimplantation development. PMID- 10840722 TI - Differential screens with subtracted PCR-generated cDNA libraries from subregions of single mouse embryos. PMID- 10840723 TI - HPLC-based mRNA differential display. PMID- 10840724 TI - Production of transgenic Drosophila. PMID- 10840725 TI - Transgenic manipulation of the sea urchin embryo. PMID- 10840726 TI - Transgenic zebrafish. PMID- 10840727 TI - Production of avian chimeras and germline transmission. PMID- 10840728 TI - Incorporation of genetically modified cells in chicken chimeras. PMID- 10840729 TI - Long-term culture of chicken blastodermal cells (CBCs) and selection of transfected CBCs using antibiotic resistance. PMID- 10840731 TI - Production of transgenic mice. PMID- 10840730 TI - Nuclear transplantation and cloning in mammals. PMID- 10840732 TI - Production of transgenic mice with yeast artificial chromosomes. PMID- 10840733 TI - Gene-targeting strategies. AB - Gene targeting in mouse ES cells is a powerful method for studying gene function in vivo. For the novice, this combination of molecular biology, specialized tissue-culture cell lines, and mouse reproductive biology can be daunting. We present a straightforward, one might say constrained, guide for novices of gene targeting to generate a null allele in large or small protein-coding genes. The method we outline has evolved from years of experience of training and advising beginners on this powerful technology. We believe that a good design for a gene targeting strategy ultimately saves time, money, and research animal lives. Once you feel comfortable with a fundamental knockout, we suggest you consider the new and exciting gene-targeting variations that can be used to address important biological questions (2). PMID- 10840734 TI - Chimeric animals and germline transmission. PMID- 10840735 TI - Conditional gene knockout using cre recombinase. PMID- 10840736 TI - Application of Cre/loxP in Drosophila. Site-specific recombination and transgene coplacement. PMID- 10840737 TI - Purification of chemokines from natural sources. PMID- 10840738 TI - Cloning of novel chemokines using a signal sequence trap method. PMID- 10840739 TI - Chemokine expression in insect cells. PMID- 10840740 TI - Expression of chemokines in Escherichia coli. PMID- 10840741 TI - Expression of chemokines in the periplasmic space of E. coli. PMID- 10840742 TI - Synthesis of chemokines. PMID- 10840743 TI - Identification of novel chemokines from expressed sequence tag databases. PMID- 10840744 TI - Purification of recombinant chemokines from E. coli. PMID- 10840745 TI - Chemokine receptor cloning. PMID- 10840746 TI - Generation of stable cell lines expressing chemokine receptors. PMID- 10840747 TI - Modified microchemotaxis assays. PMID- 10840748 TI - Transwell chemotaxis. PMID- 10840749 TI - Endothelial cell chemotaxis assays. PMID- 10840750 TI - Radiolabeled chemokine binding assays. PMID- 10840751 TI - Scintillation proximity binding assay. PMID- 10840752 TI - Calcium mobilization. PMID- 10840753 TI - Actin polymerization. PMID- 10840754 TI - Reactive oxygen release. PMID- 10840755 TI - Histamine release. PMID- 10840756 TI - Measurement of phosphoinositide 3-kinase activity. PMID- 10840757 TI - Glycosaminoglycan binding assays. PMID- 10840758 TI - CFU-A assay for measurement of the antiproliferative effects of chemokines on murine early hemopoietic progenitors. PMID- 10840759 TI - Downmodulation and recycling of chemokine receptors. PMID- 10840760 TI - Analysis of chemokine receptor endocytosis and recycling. PMID- 10840761 TI - Chemokine inhibition of HIV infection. PMID- 10840762 TI - The production of chemokine specific monoclonal antibodies. Chemokine RANTES. PMID- 10840763 TI - Monoclonal antibodies to chemokine receptors. PMID- 10840764 TI - Targeted expression of chemokines in vivo. PMID- 10840765 TI - Chemokine knockout mice. PMID- 10840766 TI - Chemokine receptor knockout mice. PMID- 10840768 TI - Murine model of allergic lung inflammation. PMID- 10840767 TI - Measurement of eosinophil accumulation in vivo. PMID- 10840769 TI - Murine models of airway inflammation. PMID- 10840770 TI - Rat models of respiratory inflammation. PMID- 10840771 TI - Murine model of crescentic nephritis. PMID- 10840772 TI - Rabbit models of pneumonia, peritoneal sepsis, and lung injury. AB - To study the mechanisms that link sepsis with ARDS, many animal models have been developed. In this chapter, a rabbit model of sepsis secondary to an intrapulmonary or intraabdominal infection has been described. One advantage of the rabbit model of sepsis is that this species produces the C-X-C chemokine, IL 8. In contrast, rodents, which are often used in studies of sepsis and ARDS, lack this important chemokine. A second advantage is the rabbit's size. This species is large enough so that the measurement of physiological parameters (e.g., mean arterial pressure, heart rate, etc.) is not difficult, but they are not so large that they require large quantities of precious reagents (e.g., recombinant proteins and MAbs). A disadvantage of the rabbit model is that there are fewer reagents (e.g., recombinant cytokines and MAbs) available for the study of inflammation in rabbits when compared to mice. PMID- 10840773 TI - Semi-permeabilized cells to study procollagen assembly. PMID- 10840774 TI - Quantitative determination of collagen crosslinks. PMID- 10840775 TI - Analysis of laminin structure and function with recombinant glycoprotein expressed in insect cells. PMID- 10840776 TI - Recombinant collagen trimers from insect cells and yeast. PMID- 10840777 TI - Eukaryotic expression and purification of recombinant extracellular matrix proteins carrying the Strep II tag. PMID- 10840778 TI - Preparation of isotopically labeled recombinant fragments of fibronectin for functional and structural study by heteronuclear nuclear magnetic resonance spectroscopy. PMID- 10840779 TI - Reconstitution of functional integrin into phospholipid vesicles and planar lipid bilayers. PMID- 10840780 TI - Confocal-FRAP analysis of ECM molecular interactions. PMID- 10840781 TI - Electron cryomicroscopy of fibrillar collagens. PMID- 10840782 TI - Atomic force microscopy measurements of intermolecular binding strength. PMID- 10840783 TI - ECM macromolecules: rotary shadowing and scanning transmission electron microscopy. PMID- 10840784 TI - Screening for mutations in cartilage ECM genes. PMID- 10840785 TI - Tissue-specific KO of ECM proteins. PMID- 10840786 TI - Homologous gene targeting to study ECM assembly. PMID- 10840787 TI - Enhancer analysis of the alpha 1(II) and alpha 2(XI) collagen genes in transfected chondrocytes and transgenic mice. PMID- 10840788 TI - Retroviral delivery of ECM genes to cells. PMID- 10840789 TI - Using self-assembled monolayers to pattern ECM proteins and cells on substrates. PMID- 10840790 TI - Methods for preparing extracellular matrix and quantifying insulin-like growth factor-binding protein binding to the ECM. PMID- 10840791 TI - Measuring interactions between ECM and TGF beta-like proteins. PMID- 10840792 TI - Using organotypic tissue slices as substrata for the culture of dissociated cells. PMID- 10840793 TI - Neuroepithelial differentiation induced by ECM molecules. PMID- 10840794 TI - Migration assays for oligodendrocyte precursor cells. PMID- 10840795 TI - Cell adhesion assays. PMID- 10840796 TI - Tissue engineering and cell-populated collagen matrices. PMID- 10840797 TI - Solid phase assays for studying ECM protein-protein interactions. PMID- 10840798 TI - Tissue engineering of cartilage. PMID- 10840799 TI - Tissue recombinants to study extracellular matrix targeting to basement membranes. PMID- 10840800 TI - Fluorescence assays to study cell adhesion and migration in vitro. PMID- 10840801 TI - Analyzing cell-ECM interactions in adult mammary gland by transplantation of embryonic mammary tissue from knockout mice. PMID- 10840802 TI - Fluid mechanics of biological surfaces and their technological application. AB - A survey is given on fluid-dynamic effects caused by the structure and properties of biological surfaces. It is demonstrated that the results of investigations aiming at technological applications can also provide insights into biophysical phenomena. Techniques are described both for reducing wall shear stresses and for controlling boundary-layer separation. (a) Wall shear stress reduction was investigated experimentally for various riblet surfaces including a shark skin replica. The latter consists of 800 plastic model scales with compliant anchoring. Hairy surfaces are also considered, and surfaces in which the no-slip condition is modified. Self-cleaning surfaces such as that of lotus leaves represent an interesting option to avoid fluid-dynamic deterioration by the agglomeration of dirt. An example of technological implementation is discussed for riblets in long-range commercial aircraft. (b) Separation control is also an important issue in biology. After a few brief comments on vortex generators, the mechanism of separation control by bird feathers is described in detail. Self activated movable flaps (= artificial bird feathers) represent a high-lift system enhancing the maximum lift of airfoils by about 20%. This is achieved without perceivable deleterious effects under cruise conditions. Finally, flight experiments on an aircraft with laminar wing and movable flaps are presented. PMID- 10840803 TI - Social hackers: integration in the host chemical recognition system by a paper wasp social parasite. AB - Obligate social parasites in the social insects have lost the worker caste and the ability to establish nests. As a result, parasites must usurp a host nest, overcome the host recognition system, and depend on the host workers to rear their offspring. We analysed cuticular hydrocarbon profiles of live parasite females of the paper wasp social parasite Polistes sulcifer before and after usurpation of host nests, using the non-destructive technique of solid-phase micro-extraction. Our results reveal that hydrocarbon profiles of parasites change after usurpation of host nests to match the cuticular profile of the host species. Chemical evidence further shows that the parasite queen changes the odour of the nest by the addition of a parasite-specific hydrocarbon. We discuss the possible role of this in the recognition and acceptance of the parasite and its offspring in the host colony. PMID- 10840804 TI - Initial homotypic cell pair adhesion in regenerating Hydra facilitates subsequent adhesion of homotypic cells. AB - In Hydra vulgaris at the level of dissociated single cells endodermal cells adhere to each other more readily than to ectodermal cells at the initial adhesion. The time required for adhesion to occur between two adjacent cells is shorter for both endodermal and ectodermal homotypic cell adhesions once the initial adhesion of the first pair of cells has been established. It is confirmed that contact of an aggregated pair with additional homotypic cells facilitates the occurrence of homotypic adhesions; heterotypic adhesions are discouraged. This suggests that adhesion of homotypic cells contributes to an increased readiness for subsequent homotypic cells to adhere. PMID- 10840805 TI - Indian-Asian relationship: mtDNA reveals more. AB - Recent studies on human mtDNA have identified continent-specific restriction enzyme sites and resultant haplo-groups among populations from different regions of the world. Such studies have helped in elaborating the models for human migrations. We have studied Indian mtDNAs to identify the recognized world ethnic elements present in it. The results presented here are based on the study of DdeI 10394 site along with the associated Asian-specific AluI 10397 site in the mtDNA sequences of the Indian samples. On examining all the related haplo-groups, this study suggests that the apparent affinities of Indians and East Asians (comprising Chinese, Japanese, Southeast Asians etc.) could be due to a proto Asiatic element present in Indians. PMID- 10840806 TI - Methane ice worms: Hesiocaeca methanicola colonizing fossil fuel reserves. AB - During a research cruise in July 1997 in the Gulf of Mexico we discovered a gas hydrate approximately 1 m thick and over 2 m in diameter which had recently breached the sea floor at a depth of 540 m. The hydrate surface visible from the submarine was considerably greater than that of any other reported hydrate. Two distinct color bands of hydrate were present in the same mound, and the entire exposed surface of the hydrate was infested (2500 individuals/m2) with 2 to 4 cm long worms, since described as a new species, Hesiocaeca methanicola, in the polychaete family Hesionidae (Desbruyeres and Toulmond 1998). H. methanicola tissue stable isotope values are consistent with a chemo-autotrophic food source. No evidence of chemo-autotrophic symbionts was detected, but geochemical data support the presence of abundant free living bacteria on the hydrate. The activities of the polychaetes, grazing on the hydrate bacteria and supplying oxygen to their habitats, appears to contribute to the dissolution of hydrates in surface sediments. PMID- 10840807 TI - Intramolecular stable isotope ratios of N2O in the tropical swamp forest in Thailand. AB - The dry-wet cycle is a common climatic phenomenon in the tropical regions of monsoon-affected Asia. An intermittent increase in N2O was clearly observed in the Bang Nara River, with a tropical swamp catchment at the beginning of rainy season of November 1997. The intramolecular distribution of isotopes clearly demonstrated that nitrification is the major process for the production of N2O. Using stable isotope data, the production mechanism of N2O in the study areas was found to vary in time and space with changes in the redox boundary along the water flow. PMID- 10840808 TI - Thermal gains through collective metabolic heat production in social caterpillars of Eriogaster lanestris. AB - We investigated thermal characteristics of aggregations of social, tent-building caterpillars of the small eggar moth Eriogaster lanestris (Lepidoptera: Lasiocampidae). The highly synchronous behavior of individuals of the colony has important consequences for their thermal ecology. Air temperature in the tent fluctuates according to the caterpillars' activity: air temperature slowly rises about 2.5-3 degrees C above the surroundings when caterpillars aggregate in the tent after feeding and decreases rapidly when the larvae leave the tent. Thermal energy can be stored for a few hours when ambient temperature drops. Experiments show that metabolic heat production sufficiently explains this effect. As even minor additional heat gain may reduce developmental time, aggregating in the tent may thus confer selective advantages under overcast weather or at night, when behavioral thermoregulation through basking is not possible. PMID- 10840809 TI - Nutrition situation in the Near East Region. PMID- 10840810 TI - The state of nutrition in Saudi Arabia. PMID- 10840811 TI - Dietary intake and nutrition related disorders in Lebanon. PMID- 10840812 TI - State of nutrition in Turkey. PMID- 10840813 TI - Food consumption patterns and nutritional problems in the Islamic Republic of Iran. PMID- 10840814 TI - The state of nutrition in Bahrain. PMID- 10840815 TI - [The new WHO classification of malignant lymphoma. After "REAL" a further step on the road to a worldwide consensus]. AB - For decades, rival classification systems for malignant lymphoma that were hardly even comparable have existed side by side: the Kiel Classification has so far been used in the German-speaking parts of Europe and the "Working Formulation" mainly in the USA. At the beginning of the 1990's it became possible to overcome this unsatisfactory situation, when pathologists in the Old and New Worlds joined forces to elaborate a new classification for lymphoma, the Revised European American Lymphoma (REAL) Classification, based on current scientific data. One new feature is the absence of a malignancy grading that is applicable to more than one entity. The system centres on the definition of distinct lymphomatous diseases that can be diagnosed with a high degree of reproducibility. Morphological and clinical characteristics are used for diagnosis, but so, consistently, are immuno-phenotypical and molecular genetic ones. An international cooperative study has confirmed the high reproducibility and clinical practicability of this tentative classification system. An international panel of pathologists appointed by the World Health Organization has updated the REAL classification, which is expected to be published in the updated version in 2001 under the title New WHO Classification. The articles that follow are based on the new WHO classification. PMID- 10840816 TI - [Lymphatic system and differentiation of B and T lymphocytes]. AB - Most malignant tumours are classified according to their cellular origins. With regard to the classification of the malignant lymphomas, however, the application of this concept was not possible for a long period. Attitudes towards this changed rapidly with the discovery and description of the various cellular components of lymphoid tissue, and with the development of methods, which reliably distinguish the cells of the lymphoid system from each other. It has become apparent that the cellular origin of most lymphomatous diseases determines their respective morphological and clinical characteristics and, therefore, represents a sound basis for the classification of most lymphomas. This principle of classifying malignant lymphomas was initially applied by the Kiel Classification. Since itsi publication, further scientific discoveries have enabled us to close the gaps in our knowledge regarding the development of lymphoid tissue and cells. These advances were incorporated in the REAL Classification, and, subsequently, in the new WHO Classification. It is obvious, therefore, that an understanding of the malignant lymphomas and their classification requires a basic understanding of the differentiation of the cells of the lymphoid system. Consequently, a description of the components, the development and of the various developmental stages of the lymphoid cells will be briefly reviewed here, serving as a basis for the following chapters, which will summarise the characteristics of the different lymphomas. As the majority of the malignant lymphomas arise from B- and T-cells, these cell types will be concentrated upon in this review. PMID- 10840817 TI - [Hodgkin lymphoma. Classification and pathogenesis]. AB - In the last few years our understanding of Hodgkin's lymphoma (HL) has enormously progressed. Molecular analysis has revealed that almost all cases of this disease are clonal B cell neoplasms, therefore the term Hodgkin's lymphoma instead of Hodgkin's disease is being proposed in the new WHO classification. Lymphocyte predominance HL (LPHL) differs in respect to morphology, immunophenotype and clinical features from the other forms of HL and represents its own distinct entity. In addition to morphologic features (nodularity, presence of L&H cells) the immuno-phenotype of tumor cells is most important in establishing a diagnosis of LPHL, and particularly in differentiating LPHL from the other forms of HL. The remaining forms of HL (nodular sclerosis, mixed cellularity, lymphocyte depletion) display a mostly identical antigen profile and similar clinical characteristics, they are therefore grouped together in the REAL classification under the heading of classical HL. Recent immuno-histological analysis have revealed that one third of HL cases, which formerly were classified as LPHL, display the immuno-phenotype of classical HL. These cases are now considered to represent examples of classical HL and termed nodular lymphocyte rich classical HL. According to retrospective clinico-pathological analysis, the biological behaviour of this newly identified form of classical HL also differs from LPHL. Differences between classical HL and LPHL also occur on the molecular level. Thus LPHL often displays ongoing mutations of the immunoglobulin genes, and the tumor cells express immunoglobulin protein and transcripts, while these characteristics are absent in classical HL. Since peripheral B cells that do not express immunoglobulins die from apoptosis, these findings imply that the regulation of apoptosis is defective in Hodgkin and Sternberg Reed cells. Several laboratories are currently working intensely to clarify the defective apoptosis pathway in HL. PMID- 10840818 TI - [The many faces of anaplastic large cell lymphoma]. AB - Fifteen years after their first description by one of the authors (HS) anaplastic large cell lymphoma (ALC-lymphoma, ALCL) now represents a generally accepted group of large cell lymphomas. Essential defining features comprise of a proliferation of large lymphoid cells with strong expression of the cytokine receptor CD30 and a characteristic growth pattern. Using molecular and clinical criteria three entities of ALC-lymphoma have been identified: primary systemic anaplastic lymphoma kinase (ALK)-positive ALC-lymphoma, primary systemic ALK negative ALC-lymphoma and primary cutaneous ALC-lymphoma. The ALK expression in the primary systemic ALC-lymphoma entity is caused by chromosomal translocations, most commonly t(2;5), and can nowadays be reliably detected by immuno-histology. ALK-positive ALC-lymphoma predominantly affects young male patients and if treated with chemotherapy has a favourable prognosis. They show a broad morphological spectrum, with the "common type", the small cell variant and the lymphohistiocytic variant being most commonly observed. The knowledge of the existence of these variants is essential in establishing the correct diagnosis. ALK-negative ALC-lymphomas occur in older patients, equally affecting both genders and have an unfavorable prognosis. The morphology and the immuno phenotype of primary cutaneous ALC-lymphoma shows an overlap with that of lymphomatoid papulosis. Both diseases have an excellent prognosis and secondary systemic dissemination is only rarely observed. The ALC-lymphomas described above derive from T cells and are generally accepted as biological entities. In contrast, large B-cell-lymphomas with anaplastic morphology are now believed not to represent an own entity but a morphologic variant of diffuse large B-cell lymphoma. Malignant lymphomas with morphological features of both Hodgkin- and ALC-lymphoma have formerly been classified as ALCL Hodgkin-like. Recent immuno histological analysis of these cases however suggests that ALCL Hodgkin-like does not represent an own lymphoma entity. Most of these cases are likely to be examples of tumor cell rich classical Hodgkin lymphoma, while a minority of these cases appear to fall either into the category of ALK-positive or ALK-negative ALC lymphoma. PMID- 10840819 TI - [Clinico-pathologic forms of peripheral T-and NK-cell lymphomas]. AB - Malignant lymphomas, originating from peripheral T or NK cells, are rare tumours in Europe and account for less than 10% of all malignant lymphomas. In this review, the salient features of the more frequently occurring entities derived from T or NK cells will be presented. Nasal NK/T cell lymphoma is mainly found in the nose and paranasal sinuses and often, but not always, display an angiocentric growth pattern leading to coagulation necrosis. The tumor cells consistently express CD56, CD2 and the EBER molecules encoded by the Epstein-Barr virus. Clonal T cell receptor gene rearrangements are often absent indicating, in the majority of cases, a derivation of these tumors from NK cells. Enteropathy-type intestinal T-cell lymphomas often arise in patients with celiac disease and have a dismal prognosis. The tumour cells express T cell antigens, CD103 and cytotoxic molecules, but are negative for CD4. Approximately 20% of the cases display CD56 mostly in combination with CD8. Recently, an early purely intraepithelial form of this tumour was identified. Histologically these cases resemble celiac disease, however the intraepithelial lymphocytes often exhibit an abnormal immunophenotype with absent CD8 and T-cell-receptor protein expression, and, they are clonal by molecular analysis. Clinically, the patients suffer from refractory sprue or ulcerative jejunitis. The prognosis is bad with the patients often dying from malnutrition or an invasive tumour-forming T-cell lymphoma. Angioimmunoblastic T cell lymphoma is defined by characteristic morphological findings (atypical lymphoid cells in part with pale cytoplasm, arborizing high endothelial venules and large FDC-meshworks) as well as clinical features (systemic symptoms, signs of a dys-regulated immune response). Peripheral T-cell lymphomas, that do not fit into a distinct entity, are classified in the REAL and the new WHO classifications as peripheral T-cell lymphomas unspecified. These display a broad morphological spectrum (including the T-cell lymphomas of different cell sizes, Lennert's lymphoma and T-zone lymphoma of the Kiel-classification) and in general are clinically aggressive. PMID- 10840820 TI - [Small cell B-cell lymphomas: guidelines for differential diagnosis]. AB - Similar to the R.E.A.L-System, the small cell B-cell lymphomas of the new WHO classification consist of chronic lymphocytic leukaemia of B cell type, mantle cell lymphoma, follicular lymphoma, lymphoplasmocytic lymphoma/immunocytoma, hairy cell leukaemia, as well as plasmacytoma. The only major difference between the WHO- and the REAL-classification is the consideration of prolymphocytic leukaemia as a single disease entity in the former system. All the above mentioned lymphomas arise from B cells of varying stages of differentiation and, therefore, often demonstrate architectural, cytological and immunophenotypic characteristics of their normal physiological counterparts. Consideration of tumour cell growth pattern, -cytology, -immunophenotype and -growth fraction, together with the presence and consistency of the reactive cell infiltrate, usually leads to categorisation of a lymphoma in the majority of cases. The molecular biological characteristics of follicular lymphoma and mantle cell lymphoma are the best defined of the small cell B-cell lymphomas. Chromosomal translocations involving the immunoglobulin heavy chain genes and the bcl-2 gene or Cyclin D1 gene, respectively, probably belong to the initial changes in a cell, which, together with several subsequent unidentified genetic alterations, lead to the development of these tumours. Although nodal small cell B-cell lymphomas are usually diagnosed at an advanced stage of the disease, the progression of the disease--with the exception of mantle cell lymphomas--is often indolent. As a result, the small cell B-cell lymphomas were previously considered as "low-grade" Non-Hodgkin lymphomas in the Kiel classification. However, since the progress of a lymphoma subtype can be heterogeneous and since mantle cell lymphomas cannot really be considered as "low-grade" tumours, "umbrella grading" of lymphomas has been discarded in the WHO classification, with emphasis being placed on grading within a lymphoma disease entity. In the following pages, the characteristics important for the diagnosis and categorisation of the small cell B-cell lymphomas will be summarised. Further, we present information regarding the molecular biological and clinical characteristics of these lymphomas. PMID- 10840821 TI - [Marginal zone lymphomas: extranodal MALT type, nodal and splenic]. AB - Marginal zone B-cell lymphomas (MCL) of extranodal, nodal and splenic origin appear to be different lymphoma entities with a similar growth pattern in the marginal zone of the B-follicles. Decisive for the detection of MCL as a distinct lymphoma entity was the "MALT concept" for lymphoid infiltrates in the gastric and intestinal mucosa as described by Isaacson et al. in the 1980's. Immunohistological stainings for the immunoglobulin light and heavy chains and molecular pathological studies of the immunoglobulin heavy chain gene configuration have subsequently confirmed the neoplastic nature of the extranodal infiltrates and differentiated marginal zone cells from mantle zone cells. In 1994, the MCL of MALT type as well as of nodal and splenic origin were included in the REAL classification and in 1998 in the new WHO classification for lymphomas. Meanwhile extranodal MCL of MALT-type have been observed in almost every organ and site of the body, by far most frequently in the gastric mucosa. Beside the typical growth pattern, lymphoepithelial lesions are a distinct diagnostic feature of extranodal MCL. Clinically, the small cell extranodal MCL show a very good prognosis with regression after treatment. As for nodal and splenic MCL, we need further studies to evaluate the prognostic aspects and to compare them with other B-cell lymphomas. The same is true for primary extranodal large B-cell lymphomas or blastic transformation to a large cell lymphoma; in these tumors the diagnosis of a MALT type lymphoma should only be made if a small cell component with MALT-specific criteria can be proved. PMID- 10840822 TI - [Large B-cell lymphomas: variants and entities]. AB - Large B-cell neoplasms represent one of the most frequent groups of non-Hodgkin lymphomas (30-40%). They are characterized by an aggressive clinical course. These lymphomas may evolve either de novo or secondary during the course of a less aggressive lymphoma. In addition to primary nodal, a primary extranodal manifestation is rather common. The neoplastic cells, even within one given case, show a broad morphological spectrum. Several findings of the last two decades have revealed that the large B-cell lymphomas represent an inhomogeneous group. This fact has been taken into account by the new WHO classification of malignant lymphomas. There are two groups identified, that of the variants and that of the subtypes. The various variants (centroblastic, immunoblastic, anaplastic, T cell/histiocyte-rich) correspond to lymphomas without reproducible discriminating criteria lacking characteristic clinical, immuno-phenotypical and genetic findings. In contrast, the primary mediastinal, the intravascular, the primary effusion and primary central nervous system lymphomas represent distinct disease entities. A number of recently described large cell lymphoma types, i.e. plasma blastic, ALK-positive and primary gastric, are included in the classification, their designation as distinct entities is still under discussion. PMID- 10840823 TI - [Diagnosis of cutaneous B-cell lymphomas]. AB - Skin is the second most common site of extranodal lymphoma. The significance of cutaneous B-cell lymphomas has been underestimated due to the lack of monoclonal antibodies and molecular genetics in former years. Cutaneous B-cell lymphomas represent a heterogeneous group of entities which show variation in histology, immunophenotype and in prognosis. The most common entities presenting in skin are follicular lymphomas, marginal zone B-cell lymphomas, and diffuse large B-cell lymphomas. The majority of primary cutaneous B-cell lymphomas have an excellent prognosis. Whereas in B-cell lymphomas with secondary spread to the skin the clinical course is dependent on the prognosis of the primary lymphoma. It is evident that primary cutaneous lymphomas are distinct from nodal lymphomas and not extranodal manifestations of their lymph-node-based counterparts. In the current review the morphologic spectrum of cutaneous B-cell lymphomas comprising primary and secondary cutaneous B-cell lymphomas is discussed according to the WHO classification scheme. PMID- 10840824 TI - An immunohistochemical study of the rete ovarii and epoophoron. AB - A study to compare the immuno-histochemical profile of the human rete ovarii, and epoophoron, with the Fallopian tube and ovarian surface epithelium was performed with 31 antibodies and antisera. A reaction was present in the epithelial cytoplasm of the rete ovarii and epoophoron of mesonephric origin, for vimentin, GFAP, cytokeratin markers, (AE1/AE3, MNF116; Cam 5.2, 34 beta E12 and for the monospecific antibodies to cytokeratins 7 and 19), heat shock protein 27, in the cell membrane for HBME-1, EMA and in the subepithelial collagen for collagen IV. Reactions were present only in the epithelium in the rete ovarii for EGFR (one case) and CA-125 (four cases). A reaction was present in the epithelium of the epoophoron only for Ber-EP-4 and S100. There was no reaction with antibodies for desmin, neurofilament protein, cytokeratins 20 or 14, actin, calretinin, E cadherin, C-erb-B2, or CEA (monoclonal and polyclonal reagents). The immuno histochemical profile of the Fallopian tube was consistent with its para mesonephric origin and that in the ovarian surface epithelium was consistent with a proposed modified mesothelial origin. This study provides an immunohistochemical profile of these structures with a large panel of commonly available antibodies and antisera, confirming and extending the findings described in previous studies. PMID- 10840825 TI - Role of 17p13.3 chromosomal region in determining p53 protein immunopositivity in human astrocytic tumors. AB - Immunohistochemistry of p53 protein is being increasingly performed as a clinical service as well as in research for prediction of tumor behavior. Although early reports suggested that p53 immunopositivity was associated with p53 gene alterations, recent evidence indicates that this is not always true. Earlier, we had demonstrated the significant association of loss of heterozygosity (LOH) of the chromosomal region 17p13.3 with higher grades of human astrocytic tumors. This was independent of the heterozygosity status of p53. LOH of p53 was taken as an indicator of p53 gene alteration, which was substantiated by sequencing a subset of the tumors. In the present study, we report that p53 immunopositivity in 40 of the same set of tumors (five could not be evaluated because of paucity of tissue) was significantly associated with LOH of 17p13.3 region (Fisher's exact two-tailed, P = 0.012; odds ratio, 12) but not with LOH of p53 (Fisher's exact two-tailed, P = 0.324; odds ratio, 2.24). This indicates that the gene(s) on the 17p13.3 region of the human chromosome may be influencing the p53 immunopositivity status of glial tumors and possibly other tumors in general. This has great implications in interpreting p53 immunohistochemistry results of biopsies of various tumor types as due to p53 mutations alone. The study thus points to a new molecular correlate for p53 immuno-positivity in tumors. PMID- 10840826 TI - HPA-1, 3, 5 genotyping to establish a typed platelet donor panel. AB - Platelet antigen typing of blood donors is not routinely performed. Usually, ABO matched platelets are transfused. In situations where the recipient has a human platelet-associated (HPA) antibody resulting in refractoriness to platelet transfusions, post-transfusion purpura or neonatal alloimmune thrombocytopenia, HPA-typed platelets may be required. Having a panel of HPA-typed donors would be helpful. This study utilised PCR-RFLP and PCR-SSP methods to determine the HPA-1, 3, 5 genotypes of 1,000 blood donors to establish a panel of HPA homozygous donors who could donate platelets for patients with anti-HPA-antibodies. The study demonstrates the HPA genotype frequencies in a Western Australian blood donor population and has identified donors who are homozygous for the "aa" and "bb" genotypes for HPA-1, 3 and 5. The Australian Red Cross Blood Service-NW Region can now use these data to develop a panel of HPA-1, 3 and 5 "aa" and "bb" homozygous platelet donors, which can be accessed for patients with HPA antibodies requiring platelet transfusion. PMID- 10840828 TI - A case of keratin-positive acute myeloid leukemia: a possible role for cytokeratin 19 as a specific epithelial marker. AB - A patient with relapsed acute myelomonocytic leukemia (AML, FAB M4) developed skin infiltration by leukemic blasts. On immunochemistry, the blasts showed "bot" positive cytoplasmic staining for cytokeratins AE1/AE3 and CAM 5.2, resembling the pattern seen in Merkel cell carcinoma of skin. However, the blasts were positive for myeloid markers and negative for cytokeratin 19 and chromogranin. Aberrant immunochemical staining can lead to misdiagnosis unless a panel of antibodies of known specificity is used in tumor diagnosis, and the clinical context is taken into account. The possible role of cytokeratin 19 as a more specific marker for epithelia than keratin cocktails is discussed. PMID- 10840827 TI - Detection of protein S deficiency: a new functional assay compared to an antigenic technique. AB - Congenital protein S (PS) deficiency is associated with increased risk of venous thrombosis. To investigate the possibility of automating PS testing with decreased turnaround time, a clotting-based functional protein S assay was evaluated and compared to an antigenic method. Samples were collected from 126 patients within 5 days of their first acute cerebral infarction, from 62 controls and from 47 consecutive samples for thrombophilia investigation. The normal range for the clotting-based kit, calculated from the results of 20 healthy controls, was 62-136% (mean +/- 2 SD). Intra- and inter-assay co-efficients of variation were < 3.0 and 10.0% respectively. There was no significant correlation between the two methods (r = 0.30, P > 0.05). Two patients had low PS antigen results with normal functional levels. Both techniques were used to compare a further group of 53 patients with defined abnormalities which included nine antigenic protein S deficiencies, five protein C deficient patients, 10 patients with a lupus anticoagulant (LA), 17 Factor V Leiden (FVL) heterozygotes, two FVL homozygotes and 10 patients on therapeutic levels of heparin. In this group we found that four of nine antigenic PS deficient patients had normal functional PS levels. The test was susceptible to the FVL mutation with four of 17 FVL heterozygotes and both of two FVL homozygotes giving low levels. One of five protein C-deficient patients also had a low functional PS result with a normal antigenic level. Normal results were obtained by both methods for all of the LA and patients on therapeutic heparin. We concluded that the automated protein S clotting assay was rapid and simple to perform but appeared to be influenced by factors other than PS deficiency. Results need to be interpreted with caution but may be useful as part of a full thrombophilia investigation. PMID- 10840829 TI - Broadsheet number 55: Diagnosis of measles virus infection in the microbiology laboratory. PMID- 10840830 TI - Broadsheet number 56: Mechanisms of fetal loss. AB - The most significant task of the pathologist examining a fetal death is, if possible, the provision of an explanation for the event, which will allow the clinical attendants to counsel the family in an informed and relevant manner. In some cases, no adequate explanation will be possible, though many conditions will be excluded and the importance of this exercise is worth emphasising. Often, however, a combination of fetal, maternal or placental conditions can be found and a full or partial explanation offered, and it is the purpose of this broadsheet to highlight some of these conditions and provide suggestions as to sources of further practical help. PMID- 10840831 TI - Test and teach. Number one hundred: Part 1. Explanation and diagnosis: intravascular lymphoma. PMID- 10840832 TI - Test and teach. Number one hundred and one: Part 1. PMID- 10840833 TI - Ultrastructure of the polymorphonuclear leucocytes in human immunodeficiency virus infection. AB - The ultrastructure of polymorphonuclear leucocytes (PMNL) was studied in 16 patients infected with the human immunodeficiency virus (HIV). PMNL were isolated from HIV-infected patients with CD4+ lymphocytes counts > 200/mm3 (without signs of active infection) (n = 12) (group 1), or < 200/mm3 (n = 4) (group 2), and from 16 healthy volunteers (group 3). Immunoelectron microscopy staining using an anti beta 2 integrin antibody (anti-CD18) was performed on PMNL from three individuals of group 2 and of three individuals of group 3, before and after incubation with N-formyl-methionyl-leucylphenylalanine (f-MLP). The radical oxygen intermediates (ROI) production of PMNL was investigated by luminol-mediated chemiluminescence. A number of ultrastructural abnormalities in PMNL were found in a higher proportion in HIV-infected patients. These were: (a) an increase in the size of the Golgi apparatus and in the number of mitochondria, and in the quantity of endoplasmic reticulum; (b) some dysplastic features including large cytoplamic vacuoles, whorl of myelin, and nuclear pockets; (c) an increase prevalence of multivesicular bodies compared with control PMNL; (d) some cylindrical confronting cisternae and tubuloreticular structures. After anti-CD18 staining, gold particles were seen on the plasma membrane and more rarely inside the cytoplasm of PMNL from each group but no decrease in this staining was noted in HIV PMNL. Incubation with f-MLP similarly increased the immunostaining of the PMNL in each group. In vitro ROI production was significantly depressed for HIV PMNL compared with control PMNL. Some ultrastructural abnormalities observed in this study could support the possibility that one of the mechanisms underlying the qualitative functional defects of PMNL from HIV-infected patients may be related to some cytopathic effect. PMID- 10840834 TI - Investigation of the -1377 polymorphism on the Apo-1/Fas promoter in systemic lupus erythematosus patients using allele-specific amplification. AB - Apoptosis mediated by the Apo-1/Fas and Fas ligand pathways has been implicated in many disorders, including autoimmunity and tumorigenesis. The recent identification of two polymorphisms on the 5' flanking region of the human Apo 1/Fas gene has provided useful markers for investigation of the genetic contribution of the Apo-1/Fas gene in these diseases. The Mval polymorphism at the -670 nucleotide position is frequent in the normal population, with 51% heterozygosity. The other polymorphism, a result of single nucleotide G-->A substitution at the -1377 position, does not create or delete any restriction enzyme digestion sites. In this paper, we describe a simple and rapid method for detecting the -1377 polymorphism by using allele-specific amplification (ASA). Using the ASA method, the -1377 polymorphism in a normal Caucasian population was characterised. Frequencies of 0.13 and 0.87 for allele A and G, respectively, were observed and the homozygosity of the mutant allele (A) was found in only 2% of the population. We subsequently examined the -1377 polymorphism in sporadic systemic lupus erythematosus (SLE) patients (n = 86). The results showed that both genotype distribution and allele frequencies in SLE patients were similar to that in controls, suggesting that the -1377 promoter polymorphism is unlikely to be associated with SLE susceptibility. The description of this rapid detection method and characterisation of the -1377 polymorphism are useful means for future genetic studies in diseases in which the Fas-mediated apoptosis may be involved. PMID- 10840835 TI - An advanced digital image-capture computer system for gross specimens: a substitute for gross description. AB - The description of macroscopic appearances of surgically excised specimens together with the sites of specimen sampling form an important component in the documentation of a histopathology specimen. Unfortunately, accuracy of description depends on the vocabulary and descriptive prowess of the pathologist which means that the result can be variable. Transcription of the dictated word also takes time and involves typists. We developed a user-friendly, high resolution image capture system that will supplement word description of macroscopic specimens and has the potential to replace it completely. It also provides the opportunity of revisiting traditional methods of documenting specimens with words, allowing the production of more relevant and user-friendly reports containing information relevant to clinical management and supplemented by high-resolution digital images. The accompanying Windows-based software has capabilities of generating the entire histopathology report and allows rapid retrospective searches through any one of several common search parameters. The stored images form a powerful database for teaching and research. Connection to remote transmission facilities allows teleconferencing and telepathology consultations. PMID- 10840836 TI - Ovarian histoplasmosis in systemic lupus erythematosus. AB - Histoplasma capsulatum is a pathogenic fungus endemic to North, Central, and South America. Histoplasmosis is primarily acquired by inhalation and in immunocompetent hosts infection is generally limited to the lungs. Histoplasmosis may disseminate systemically in hosts with defective cell-mediated immunity or massive inoculation. Systemic lupus erythematosus (SLE) is an autoimmune disorder associated with multiple primary and drug-related immunological defects that predispose patients to infections. Disseminated histoplasmosis has only rarely been described in association with SLE. We describe a 32-year-old woman with SLE who had a 2-year history of irregular menses and a confirmed anovulatory state, secondary to ovarian histoplasmosis. The ovarian histoplasmosis was discovered incidentally at surgery for a persistent colo-cecal fistula, which had developed 6 months earlier and was originally associated with an Actinomyces and Fusobacterium pelvic abscess. The patient had no evidence of active pulmonary histoplasmosis and her disseminated histoplasmosis likely resulted from re activation of latent disease. This is the first description of disseminated histoplasmosis presenting as ovarian dysfunction in a patient with SLE. PMID- 10840837 TI - Ectopic hamartomatous thymoma: a case study and review of the literature. AB - Ectopic hamartomatous thymoma is a rare and distinctive tumor found in the deep soft tissues of the neck, which is characterized histopathologically by a mixture of spindle, epithelial, and adipose cell elements. We present a case of this lesion occurring in a 39-year-old male. The characteristic histochemical and immunohistochemical findings of these tumors are demonstrated. In addition, review of the reported histological, immunohistochemical and ultrastructural findings is presented together with a table of clinical findings in the tumors so far described. A brief discussion of theories of histogenesis and possible differential diagnosis is included. PMID- 10840838 TI - Basaloid squamous carcinoma in the liver. AB - This report concerns a 46-year-old female who presented with 3 months of abdominal pain and underwent a right hemi-hepatectomy for a 27 x 25 x 15 cm, centrally necrotic tumor that showed histological, immunohistochemical and ultrastructural features typical of a basaloid squamous carcinoma (BSC). A primary tumor at another site was not diagnosed and she died of disease 2 years later after several intra-abdominal recurrences. The entity of BSC was first described in 1986 and is a rare, poorly differentiated variant of squamous cell carcinoma occurring in various sites including the upper aerodigestive tract, esophagus, lung, anus, cervix and thymus. It has never been reported arising in the liver. It has characteristic histological, immunohistochemical and ultrastructural features and is associated with a poor prognosis. Whilst no other primary tumor was diagnosed, it is not possible to substantiate that this is a primary hepatic tumor in the absence of an autopsy examination to exclude an occult malignancy in another site. PMID- 10840839 TI - Pasteurella multocida: a case report of bacteremic pneumonia and 10-year laboratory review. AB - Pasteurella multocida is a normal oral commensal in animals. Animal bites are often complicated by severe wound infection due to P. multocida, but systemic infection is rare. We report a patient with bacteremic pneumonia successfully treated with ceftriaxone and ciprofloxacin. We also review the clinical isolates of P. multocida reported by a major teaching hospital laboratory over a 10-year period. There were 23 patients, comprising the present case, 17 patients with wound infections following animal bites, one case of neonatal meningitis and associated maternal vaginal carriage of P. multocida, and three sputum isolates of doubtful significance. PMID- 10840841 TI - An alternative to the "reference range" for reporting urinary sodium and potassium, and blood lipids. PMID- 10840840 TI - Extraskeletal osteosarcoma of the scalp. AB - A rare case of extraskeletal osteosarcoma of the scalp in a 56-year-old woman is described. At presentation she was found to have an 8-cm diameter, tender, firm, exophytic scalp tumor. MRI scan confirmed absence of underlying skeletal origin and showed extension along the subcutaneous plane. The tumor was excised and the patient received post-operative chemotherapy. Histologically, the tumor showed classical features of an osteogenic osteosarcoma with focal fibroblastic areas. In addition, there were rhabdoid cells present, which showed paranuclear cytoplasmic immunoreactivity for epithelial membrane antigen. The patient developed metastatic disease 6 months after surgical excision. PMID- 10840842 TI - [The Internet as a communications medium in radiology]. PMID- 10840843 TI - [Radiology and the Internet. The development, foundations and technical prerequisites]. AB - As a new communication media the Internet has revolutionized the global exchange of information which is one of the major sources of progress in medicine. This article reviews the history of the Internet's evolution and describes some fundamental technical concepts. The importance of Internet services for everyday professional work is examined with special consideration of radiology. Special emphasis is laid on the current technical limitations that have delayed a broad practical implementation of tele-medical applications. As a highly innovative medical specialty, radiology has always been at the forefront of research and development in the field of clinical Internet applications. By taking a look at recent innovations, the impact of the Internet on the radiologist's working environment of the future is highlighted. PMID- 10840844 TI - [Internet communication in radiology]. AB - E-mail is an Internet service that can be used for sending messages and binary files between individuals as well as for participating in discussion groups. For sending and receiving these types of messages, the users must use either a dedicated e-mail client or one of the several mailing facilities of the World Wide Web. The news-groups enable like-minded people to discuss subjects on a group-wide basis, but access is generally not limited, and the participants cannot be selected. CONCLUSION: The objective of this paper is to give radiologists an introduction to using e-mail, mailing lists and news-groups, the three most important communication services of the Internet. The function of these services is explained, and the advantages of implementing them in a radiology practice are discussed. Potential problems and concerns including security matters are highlighted, and ways in which they can be resolved are suggested. PMID- 10840845 TI - [Radiological information searching on the Internet: data banks, search machines and intelligent agents]. AB - The Internet plays an important role in a growing number of medical applications. Finding relevant information is not always easy as the amount of available information on the Web is rising quickly. Even the best search engines can only collect links to a fraction of all existing Web pages. In addition, many of these indexed documents have been changed or deleted. The vast majority of information on the Web is not searchable with conventional methods. New search strategies, technologies and standards are combined in intelligent search agents (ISA) and robots, which can retrieve desired information in a specific approach. CONCLUSION: The article describes differences between ISAs and conventional search engines and how communication between agents improves their ability to find information. Examples of existing ISAs are given and the possible influences on the current and future work in radiology is discussed. PMID- 10840846 TI - [Researching and publishing on the Internet]. AB - The popularity of Internet is rapidly growing in all fields, and Radiology is no exception. Thousands of websites with radiological data have been enthusiastically created and are now available on-line, competing with the huge number of sites offering any kind of information or commercial services. After the pioneering experiences of the early Internet years, now time has come to critically analyze the opportunities and the risks of using Internet as a channel for distributing radiological knowledge. PMID- 10840847 TI - [Internet technology for clinical applications in a digital radiography department]. AB - PROBLEM: To provide an overview and to assess the clinical feasibility of Internet technology-based systems for hospital-wide image and report distribution as well as for video conferencing. METHODS: The paper describes the theoretical concept behind, the various technical approaches and the experience gained from different systems. RESULTS: Image and report distribution: Advantages include the universal availability of images and reports inside and outside hospitals; ease of use; security features; image and report integration; cost savings by reducing support and training efforts and by optimising available hardware. The main critical issues are performance and workflow integration with RIS and PACS. Video conferencing: main advantages are the standardised, software-based approach and the low investments for hard- and software. Depending on the desired usage the communication performance can be seen as inappropriate. CONCLUSION: Today, Internet technology-based systems appear to satisfy the main clinical needs in radiology. The mentioned drawbacks could be eliminated by means of modified software implementation and focused standardisation efforts. Considering the numerous advantages of these systems a further distribution can be expected for the future. PMID- 10840848 TI - [Training and continuing education via the Internet]. AB - A world-wide homogeneous level of knowledge for all members of an expert community is the proposed aim of academic education. The potential of the World Wide Web (WWW) with respect to under- and postgraduate education maybe accomplished if its use will become a definite part of cultural skills. Additionally, the process of simple collection of information should be added by the process of gaining knowledge and experience. Adaptation to the target group and a high degree of inter-activity are the two main characteristics of high quality technology-based learning materials. Considering both most of the web resources show deficits. Instructional design as well as information technology have means to overcome these deficits. With respect to the didactic principles of materials for beginners and advanced students, the architecture of computerised learning programs plays a significant role. However, costs to develop high-level learning materials is extremely high. Therefore it is necessary, to use synergetic effects with other digitized clinical and scientific materials. PMID- 10840849 TI - [Security aspects on the Internet]. AB - AIM OF THE STUDY: Is it possible to use the Internet as a secure media for transport of telemedicine? Which risks exist for routine use? In this article state of the art methods of security were analysed. Telemedicine in the Internet has severe risks, because patient data and hospital data of a secure Intranet can be manipulated by connecting it to the Web. CONCLUSIONS: Establishing of a firewall and the introduction of HPC (Health Professional Card) are minimizing the risk of un-authorized access to the hospital server. HPC allows good safety with digital signature and authentication of host and client of medical data. For secure e-mail PGP (Pretty Good Privacy) is easy to use as a standard protocol. Planning all activities exactly as well as following legal regulations are important requisites for reduction of safety risks in Internet. PMID- 10840850 TI - [The radiological diagnosis of an intraorbital bullet projectile]. AB - Beside conventional X-ray examination, ultrasound and computer tomographic examination are accepted radiologic techniques to assess the extend of damage of intraorbital injuries. With the exception of a few reports in the literature there is agreement that intraorbital metallic foreign bodies and gun-shot bullets represent a contraindication for magnetic resonance (MR) imaging examination because of artificial imaging side effects and the potential of secondary dislocation of the metallic foreign body due to ferromagnetism. By means of the case reported here it is shown that this general opinion is not universally valid. Depending on the elemental metallic composition of foreign bodies and gun shot bullets it is demonstrated that MR examination allows high-resolution illustration and in some cases is advantageous compared with other radiologic imaging techniques. PMID- 10840851 TI - [An accidental finding after a traffic accident: an intra- and suprasellar space occupying lesion]. PMID- 10840852 TI - [Chronic inflammatory intestinal diseases: Crohn's disease and ulcerative colitis. 2. The imaging procedure in ulcerative colitis and the therapy of Crohn's disease and ulcerative colitis]. PMID- 10840853 TI - Mobile chest radiography: improving image quality. AB - Mobile examinations present special challenges yet require the same diagnostic quality as exams performed in the radiology department. This article describes a study designed to identify common errors in mobile chest radiography. The authors identify 3 areas of particular concern: rotation, anatomy not centered to the film and distortion. Possible solutions are suggested and discussed. PMID- 10840854 TI - Provider efforts to increase mammography screening. AB - This article examines barriers to mammography screening and reviews the professional literature on provider interventions to increase screening. Interventions appropriate for radiologic technologists are evaluated and discussed in an effort to expand R.T.s' role in improving mammography screening rates. PMID- 10840855 TI - The role of ultrasound in breast imaging. AB - Sonography has become an important tool for evaluating breast abnormalities, including inflammatory processes, hematomas, ductal ectasia, cysts and solid lesions. This article describes protocols for ultrasound imaging of the breast and the sonographic appearance of common breast conditions, with particular attention to differentiating benign from malignant lesions. Ultrasound's role in interventional breast procedures and radiation therapy planning also are discussed. PMID- 10840856 TI - Radiologic assessment of schizophrenia. AB - Schizophrenia is a debilitating psychotic disorder. Neuroimaging, particularly magnetic resonance and computed tomography, plays a central role in describing schizophrenia and in characterizing its progression from mild childhood symptoms to adult psychosis. This article introduces readers to schizophrenia, its precursors, risk factors, progressive symptomatology and imaging techniques used to assess the disease. PMID- 10840857 TI - A variation for imaging the acetabulum. PMID- 10840858 TI - Competency, objectives and outcomes. AB - As we look at the literature from the 1970s and the present, it seems we have come full circle with regard to our understanding of objectives, competencies and outcomes. Our guidelines for writing them have become very complex. During the past few decades, health professions educators often have lost sight of the original intent and focused on the process of developing and writing objectives and competency statements. The process is now so complex that it is almost impossible to write a simple, understandable objective or competency statement. Perhaps that is why the "outcome" term was born. Outcome statements are really fairly broad competency statements. If we focus on the real purpose of objectives and make them simple, they become easier to evaluate. In the next article, we will explore methods for evaluating competence in the clinical setting. PMID- 10840859 TI - One light to leak. PMID- 10840861 TI - R.T.'s invention facilitates positioning. PMID- 10840860 TI - A technique for lower limb lengthening. PMID- 10840862 TI - Reader clarifies diagnostic criteria for osteoporosis. PMID- 10840863 TI - Folic acid and the prevention of disease. Report of the Committee on Medical Aspects of Food and Nutrition Policy. PMID- 10840864 TI - Alcohol and the athlete. AB - Alcohol and the athlete have been linked together since ancient times. It continues to be the most commonly consumed drug among the athletic population. Alcohol use carries significant potential adverse effects for both the health and welfare of the individual. It is suggested that alcohol related problems may be more prevalent in the athletic population due to their risk taking mentality and the age profile of athletes (18- to 24-year-old males). Alcohol consumption also appears to have a causative effect in sports related injury, with an injury incidence of 54.8% in drinkers compared with 23.5% in nondrinkers (p < 0.005). This may be due in part to the hangover effect of alcohol consumption, which has been shown to reduce athletic performance by 11.4%. Alcohol is a potentially lethal drug and is a banned substance for certain Olympic sports. Education is the cornerstone for appropriate social use of this drug. Athletes and coaches need to be aware of the sports related adverse effects of alcohol consumption and its role in sports injury and poor physiological performance. It is recommended that alcohol should be avoided by the serious athlete. PMID- 10840865 TI - Exercise following myocardial infarction. Current recommendations. AB - Cardiac rehabilitation services are comprehensive long term programmes designed to limit the physiological and psychological effects of cardiovascular disease (CVD), control cardiac symptoms and reduce the risk of subsequent CVD events by stabilising or partially reversing the underlying atherosclerosis process through risk factor modification. Exercise training is the cornerstone of such programmes. Ideally, exercise conditioning or training for the stable cardiac patient should include a combination of cardiorespiratory endurance (aerobic) training, arm exercises and muscular conditioning resistance (strength) training. Flexibility exercises should also be performed, usually as part of the warm-up and cool-down routines preceding and following endurance and strength training. This review discusses the potential physiological, psychological and health benefits of regular exercise and provides guidelines for exercise training for the rehabilitation of post-myocardial infarction patients following hospitalisation. PMID- 10840866 TI - Neuromuscular and hormonal factors associated with knee injuries in female athletes. Strategies for intervention. AB - Female athletes who participate in jumping and cutting sports are 4 to 6 times more likely to sustain a serious knee injury than male athletes participating in the same sports. More than 30,000 serious knee injuries are projected to occur in female intercollegiate and high school athletics in the US each year. The majority of these injuries occur by non-contact mechanisms, most often during landing from a jump or making a lateral pivot while running. Knee instability, due possibly to decreased neuromuscular strength and coordination or increased ligamentous laxity, may underlie the increased incidence of knee injury in females. Neuromuscular training can significantly increase dynamic knee stability in female athletes. Female sex hormones (i.e. estrogen, progesterone and relaxin) fluctuate radically during the menstrual cycle and are reported to increase ligamentous laxity and decrease neuromuscular performance and, thus, are a possible cause of decreases in both passive and active knee stability in female athletes. Oral contraceptives stabilise hormone levels during the menstrual cycle and may function to either passively or actively stabilise the knee joint. The long term objective of clinicians and researchers should be to determine the factors that make women more susceptible than men to knee ligament injury and to develop treatment modalities to aid in the prevention of these injuries. The immediate objectives of this review are to examine how female and male athletes differ in neuromuscular and ligamentous control of the lower extremity. The review will examine the effects of neuromuscular training on knee stability. The effects of female hormone levels and oral contraceptives on neuromuscular control of the female athletes' knee will also be discussed. PMID- 10840869 TI - Differential effects of 6-DMAP, olomoucine and roscovitine on Xenopus oocytes and eggs. AB - The effects of the new cyclin-dependent kinase inhibitors, roscovitine and olomoucine, on oocytes and eggs of Xenopus laevis were investigated and compared with those of 6-dimethylamino purine (6-DMAP). The inhibitory properties of 6 DMAP, olomoucine and roscovitine towards p34cdc2-cyclin B isolated from Xenopus eggs revealed K-IC50 values of 300, 40 and 10 microM respectively. The three compounds inhibited progesterone-induced maturation with M-IC50 values of 200, 100 and 20 microM. These values were consistent with the K-IC50 values but the ratio M-IC50/K-IC50 was higher for roscovitine and olomoucine than for 6-DMAP. The disappearance of spindle and condensed chromosomes without pronucleus formation was observed when 1 mM 6-DMAP was applied for 4 h at germinal vesicle breakdown or at metaphase II, whereas no effect was observed using 1 mM olomoucine or 50 microM roscovitine. Changes in the electrophoretic mobility of p34cdc2 and erk2 were observed only in homogenates of matured oocytes or eggs exposed for 4 h to 1 mM 6-DMAP. When the drugs were microinjected into matured oocytes, olomoucine (100 microM) and roscovitine (50 microM) induced pronucleus formation more efficiently than did 6-DMAP (100 microM). Taken together, these results demonstrate that Xenopus oocytes possess a lower permeability to olomoucine and roscovitine and that these new compounds are suitable for in vivo studies after germinal vesicle breakdown provided they are microinjected. PMID- 10840868 TI - Functional instability following lateral ankle sprain. AB - Lateral ankle sprain (LAS) is an extremely common athletic injury. Despite extensive clinical and basic science research, the recurrence rate remains high. Functional instability (FI) following LAS is hypothesised to predispose individuals to reinjury because of neuromuscular deficits which result following injury. This paper provides an overview of the potential causes of FI which may manifest themselves clinically. The theoretical explanations of FI are discussed, as are implications for assessment and treatment of FI following LAS. When LAS occurs, structural damage not only occurs to the ligamentous tissue, but also to the nervous and musculotendinous tissue around the ankle complex. While injury to the ligaments may result in laxity of the joints of the ankle complex, neuromuscular deficits are also likely to occur due to the injury to the nervous and musculotendinous tissue. These neuromuscular deficits may be manifested as impaired balance, reduced joint position sense, slower firing of the peroneal muscles to inversion perturbation of the ankle, slowed nerve conduction velocity, impaired cutaneous sensation, strength deficits and decreased dorsiflexion range of motion. Additionally, the abnormal formation of scar tissue after injury may lead to sinus tarsi syndrome or anterolateral impingement syndrome, which may also lead to FI of the ankle complex. Assessment of patients with LAS must address not only joint laxity and swelling, but should include examination for neuromuscular deficits as well. The treatment and rehabilitation goals must also address restoration of neuromuscular function, as well as restoration of mechanical stability to the injured joints. PMID- 10840870 TI - A non-invasive method for measuring preimplantation embryo physiology. AB - The physiology of the early embryo may be indicative of embryo vitality and therefore methods for non-invasively monitoring physiological parameters from embryos could improve preimplantation diagnoses. The self-referencing electrophysiological technique is capable of non-invasive measurement of the physiology of individual cells by monitoring the movement of ions and molecules between the cell and the surrounding media. Here we use this technique to monitor gradients of calcium, potassium, oxygen and hydrogen peroxide around individual mouse preimplantation embryos. The calcium-sensitive electrode in self referencing mode identified a region of elevated calcium concentration (approximately 0.25 pmol) surrounding each embryo. The calcium gradient surrounding embryos was relatively steep, such that the region of elevated calcium extended into the medium only 4 microns from the embryo. By contrast, using an oxygen-sensitive electrode an extensive gradient of reduced dissolved oxygen concentration was measured surrounding the embryo and extended tens of micrometres into the medium. A gradient of neither potassium nor hydrogen peroxide was observed around unperturbed embryos. We also demonstrate that monitoring the physiology of embryos using the self-referencing technique does not compromise their subsequent development. Blastocyts studied with the self referencing technique implanted and developed to term at the same frequency as did unexamined, control embryos. Therefore, the self-referencing electrode provides a valuable non-invasive technique for studying the physiology and pathophysiology of individual embryos without hindering their subsequent development. PMID- 10840871 TI - The distribution and requirements of microtubules and microfilaments in bovine oocytes during in vitro maturation. AB - Microtubules and microfilaments are major cytoskeletal components and important modulators for chromosomal movement and cellular division in mammalian oocytes. In this study we observed microtubule and microfilament organisation in bovine oocytes by laser scanning confocal microscopy, and determined requirements of their assembly during in vitro maturation. After germinal vesicle breakdown, small microtubular asters were observed near the condensed chromatin. The asters appeared to elongate and encompass condensed chromatin particles. At the metaphase stage, microtubules were observed in the second meiotic spindle at the metaphase stage. The meiotic spindle was a symmetrical, barrel-shaped structure containing anastral broad poles, located peripherally and radially oriented. Treatment with nocodazole did not inhibit germinal vesicle breakdown. However, progression to metaphase failed to occur in oocytes treated with nocodazole. In contrast, microfilaments were observed as a relatively thick uniform area around the cell cortex and overlying chromatin following germinal vesicle breakdown. Treatment with cytochalasin B inhibited microfilament polymerisation but did not prevent either germinal vesicle breakdown or metaphase formation. However, movement of chromatin to the proper position was inhibited in oocytes treated with cytochalasin B. These results suggest that both microtubules and microfilaments are closely associated with reconstruction and proper positioning of chromatin during meiotic maturation in bovine oocytes. PMID- 10840867 TI - The thermophysiology of uncompensable heat stress. Physiological manipulations and individual characteristics. AB - In many athletic and occupational settings, the wearing of protective clothing in warm or hot environments creates conditions of uncompensable heat stress where the body is unable to maintain a thermal steady state. Therefore, special precautions must be taken to minimise the threat of thermal injury. Assuming that manipulations known to reduce thermoregulatory strain during compensable heat stress would be equally effective in an uncompensable heat stress environment is not valid. In this review, we discuss the impact of hydration status, aerobic fitness, endurance training, heat acclimation, gender, menstrual cycle, oral contraceptive use, body composition and circadian rhythm on heat tolerance while wearing protective clothing in hot environments. The most effective countermeasure is ensuring that the individual is adequately hydrated both before and throughout the exercise or work session. In contrast, neither short term aerobic training or heat acclimation significantly improve exercise-heat tolerance during uncompensable heat stress. While short term aerobic training is relatively ineffective, long term improvements in physical fitness appear to provide some degree of protection. Individuals with higher proportions of body fat have a lower heat tolerance because of a reduced capacity to store heat. Women not using oral contraceptives are at a thermoregulatory disadvantage during the luteal phase of the menstrual cycle. The use of oral contraceptives eliminates any differences in heat tolerance throughout the menstrual cycle but tolerance is reduced during the quasi-follicular phase compared with non-users. Diurnal variations in resting core temperature do not appear to influence tolerance to uncompensable heat stress. PMID- 10840872 TI - The effects of cell size and ploidy on cell allocation in mouse chimaeric blastocysts. AB - In a previous study of mouse tetraploid<-->diploid chimaeric blastocysts, tetraploid cells were found to be more abundant in the trophectoderm than the inner cell mass (ICM) and more abundant in the mural trophectoderm than the polar trophectoderm. This non-random allocation of tetraploid cells to different regions of the chimaeric blastocyst may contribute to the restricted tissue distribution seen in post-implantation stage tetraploid<-->diploid chimaeras. However, the tetraploid and diploid embryos that were aggregated together differed in several respects: the tetraploid embryos had fewer cells and these cells were bigger and differed in ploidy. Each of these factors might underlie a non-random allocation of tetraploid cells to the chimaeric blastocyst. A combination of micromanipulation and electrofusion was used to produce two series of chimaeras that distinguished between the effects of cell size and ploidy on the allocation of cells to different tissues in chimaeric blastocysts. When aggregated cells differed in cell size but not ploidy, the derivatives of the larger cell contributed significantly more to the mural trophectoderm and polar trophectoderm than the ICM. When aggregated cells differed in ploidy but not cell size, the tetraploid cells contributed significantly more to the mural trophectoderm than the ICM. In both experiments the contributions to the polar trophectoderm tended to be intermediate between those of the mural trophectoderm and ICM. These experiments show that both the larger size and increased ploidy of tetraploid cells could have contributed to the non-random cell distribution that was observed in a previous study of tetraploid<-->diploid chimaeric blastocysts. PMID- 10840873 TI - In vitro parthenogenesis of mouse oocytes under several experimental conditions. AB - Although the in vitro fertilisation index is a parameter commonly employed to investigate sperm functional activity, little attention has been given to the occurrence of parthenogenesis. The purpose of this study was to study at 6 h or 22 h incubation: (a) the cleavage-related events that occur in in vitro incubated mouse oocytes, in the absence (parthenogenesis) or presence of homologous spermatozoa; (b) the effect of mineral oil, commonly used in in vitro fertilisation assays; (c) the effect of piroxicam, a prostaglandin synthesis inhibitor, on the parthenogenetic rate; and (d) the influence on parthenogenesis of spontaneous loss of the cumulus oophorus coat during incubation. Under the experimental conditions employed, there was parthenogenetic activation and activation due to fertilisation. Both increased in a time-dependent manner. The mineral oil enhanced the parthenogenetic rate at 22 h incubation. However, it did not have any effect when the oocytes were inseminated. Since we can not discriminate how much of this activation was due to fertilisation and how much to parthenogenesis we must be very careful with this comparison. Piroxicam 10(-8) M did not show any effect on the mouse oocyte parthenogenetic rate at neither 6 h or 22 h incubation. Our results suggest that oocyte susceptibility to spontaneous parthenogenetic activation may be modified by the presence of the cumulus and corona radiata cells. In conclusion, we consider that further rigorous studies on these influences are necessary in order to confer more reliability on the results. PMID- 10840874 TI - Further evidence that sperm nuclear proteins are necessary for embryogenesis. AB - We have recently presented evidence that the structural integrity of the mouse sperm nuclear matrix may be necessary for the proper unpackaging of sperm DNA for participation in embryogenesis. It is likely that the sperm nuclear matrix contributes to the organisation of the sperm DNA and its disturbance can seriously damage the paternal genome or its expression. In this work, we confirm our previous data and further suggest that even very subtle changes in the sperm nuclear structure may have a significant impact on embryo development. As reported previously, dithiothreitol (DTT) in the presence of an ionic detergent, ATAB, destabilized the nuclear matrix as measured by the halo assay, and oocytes injected with these nuclei failed to develop. We also discovered that omitting the protease inhibitor PMSF from the buffers used to extract spermatozoa prevented sperm injected into oocytes from participating in development. The organization of DNA into loop domains by the nuclear matrix in these nuclei appeared normal, as measured by the halo assay. Oocytes injected with sperm nuclei that had been washed with ATAB in the presence of phenylmethylsulphonyl fluoride (PMSF) but in the absence of DTT resulted in live births. Neither DTT treatment nor the absence of PMSF would be expected to disrupt the integrity of the paternal DNA. The data therefore suggest that even very subtle alterations in the structural proteins of the nucleus are enough to deprive sperm DNA of the ability to contribute to embryonic development. PMID- 10840875 TI - Apoptosis in the early bovine embryo. AB - Cell death occurs during early development in vivo and in vitro, although little is known about the mechanism of blastomere death and the relation to embryonic loss. Apoptosis, characterised by chromatin condensation, DNA fragmentation and membrane blebbing, occurs without damage to surrounding cells in contrast to necrosis. Bovine oocytes and in vitro fertilised embryos (total n = 449) were analysed for (1) DNA fragmentation using terminal deoxynucleotidyl transferase mediated dUTP nick end labelling (TUNEL) and (2) morphological features of apoptosis. TUNEL labelling was detected in immature and mature oocytes (7%, n = 57 and 23%, n = 60, respectively), and at least one cell of 8- to 16-cell embryos (5%, n = 57), morulae/early blastocysts (79%, n = 39) and expanded/hatched blastocysts (100%, n = 48). In contrast, TUNEL labelling was not detected in zygotes (n = 61), 2-cell embryos (n = 46) or 3- to 7-cell embryos (n = 81). Chromatin condensation, nuclear fragmentation, absence of neighbouring cell destruction and extrusion of cells was frequent among advanced stage embryos. Although not detected during early cleavage under standard conditions, TUNEL labelling indicative of apoptosis was induced by treatment with 10 microM staurosporine for 30 h in 95% of cleavage stage embryos (n = 59). Determination of the expression and localisation of the p53 tumour suppressor gene using reverse transcription polymerase chain reaction and whole-mount immunofluorescence revealed that although p53 transcripts were present throughout early development, nuclear localisation of p53 protein could not be detected in any blastocyst suggesting p53-independent apoptosis. This study has shown that apoptosis is dependent on embryonic developmental stage after standard culture. This suggests that bovine embryos become more capable of accommodating damaged or abnormal cells as development proceeds. PMID- 10840876 TI - Optimisation of porcine oocyte activation following nuclear transfer. AB - Experiments were conducted to examine the effects of (a) different activation methods, (b) incubation time in calcium-free medium and (c) bisbenzimide staining on the activation and subsequent development of pig oocytes. Oocytes were matured in vitro and activated by one of the following methods: combined thimerosal/dithiothreitol (DTT) treatment, calcium ionophore A23187 treatment followed by incubation in the presence of 6-dimethylaminopurine (6-DMAP), electroporation, and electroporation followed by incubation with cytochalasin B. There were no significant differences in the activation rate (ranging from 70.0% to 88.3%) and the percentage of cleaved embryos after activation (ranging between 48.8% and 58.8%) among the four treatment groups (p < 0.05). The rate of development of the blastocyst stage in oocytes activated by thimerosal/DTT (10.0%) or electroporation followed by cytochalasin B treatment (12.3%) was significantly higher (p < 0.05) than in the group activated with A23187/6-DMAP (2.5%). Both the activation rate and the rate of blastocyst formation in oocytes that were incubated in Ca(2+)-free medium for 8 h before thimerosal/DTT activation were significantly lower (p < 0.05) than in those incubated for 0, 1 or 4 h. Intracellular Ca2+ measurements revealed that the Ca2+ homeostasis in these oocytes were severely altered. Staining of oocytes with 5 micrograms/ml bisbenzimide for 2 h decreased the quality of blastocysts and increased the rate of degenerated embryos at day 6. Two activation protocols (thimerosal/DTT and electroproation) were used for activation after nuclear transfer; the rate of nuclear formation did not differ in the oocytes activated by the two different methods. PMID- 10840877 TI - Correlation between centromere and chromosome length in human male pronuclear chromosomes: ultrastructural analysis. AB - Ultrastructural and morphometric analyses of centromeric regions by scanning and transmission electron microscopy have been performed in chromosomes from male pronuclei obtained by heterologous fertilisation of hamster oocytes with human spermatozoa. In 1308 of 1323 chromosomes analysed, the primary constriction showed a defined biconcave constriction of variable length (0.56-1.34 microns) and constant width (0.64-0.7 micron). A positive correlation was observed between centromeric length and chromosome length. In some chromosomes, the primary constriction appears as decondensed regions of variable length (1.6-2.51 microns) composed of chromatin fibres with a minimum diameter of 30 nm. PMID- 10840878 TI - Zygotic and embryonic gene expression in cow: a review of timing and mechanisms of early gene expression as compared with other species. AB - Early embryonic development is largely dependent on maternal RNAs and proteins synthesised during oogenesis. Zygotic transcription is an essential event that occurs at a species-specific time after fertilization. In the absence of zygotic transcription the embryo dies since it can no longer support requirements for successful embryo development. Molecular genetics of gene expression during early embryogenesis, especially in the bovine species, remain one of the unsolved questions in modern biology. Earlier studies suggested that embryonic transcription in cattle begins at the late 4-cell or 8-cell stage. However, more recent studies suggest that bovine zygotes and 2-cell embryos are both transcriptionally and translationally active. Moreover, changes in chromatin structure due to acetylation of core histones and DNA replication play important roles in the regulation of zygotic/embryonic gene expression. This review will summarise results of recent studies about the timing and mechanisms of zygotic/embryonic gene expression in cattle. In addition, terminology in the literature regarding gene expression during early embryogenesis will be clarified. These terminologies include: 'zygotic/embryonic gene expression', 'maternal to embryonic transition in control of development (MET)' and 'zygotic/embryonic genome activation (ZEGA)'. PMID- 10840879 TI - Should Hispanic persons from different countries of origin be combined into one sample for analysis? AB - Hispanic respondents to 21 nationwide surveys were grouped on the basis of country of origin into Mexican Americans (n = 402), Puerto Ricans (n = 119), and Other Latin Americans (n = 105). Analysis showed no significant over-all differences among subgroups. No significant subgroup differences in work attitudes were found for women and only one for men. Analysis of subgroup attitudes across age, education, occupation, family income, and personal income also gave no significant differences. These results provide tentative evidence that Hispanic respondents from different countries of origin may be aggregated into a single sample for analysis. There may be no more reason to analyze their responses separately by country of origin than there is to do so for persons of European or African origin. PMID- 10840880 TI - Coefficient alpha: some terminological ambiguities and related misconceptions. AB - Three basic concepts in psychometric theory are homogeneity, consistency, and stability. These terms, however, are not singular in meaning as used in the literature and as a result often lead to confusion. The pluralities involved are discussed and alternatives are recommended. PMID- 10840881 TI - Do sex and perception of immediate stress affect optimism? AB - The purpose was to assess differences in mean optimism scores of men and women of low immediate stress (23 men, 28 women) and high immediate stress (11 men, 34 women). Using factorial analysis of variance, mean differences on the Coping Resources Inventory, Cognitive Domain, were not significant for sex, immediate stress, or their interaction. PMID- 10840882 TI - Cultural similarities in psychological reactions to infertility. AB - A 74-item especially developed and validated questionnaire for Iranian culture was administered three times to 37 Iranian infertile couples and 10 fertile couples acting as control group, namely, initial consultation, during treatment, and on completion of treatment. In view of parallel data also reported for Western patients interest centered on the results of two out of the five factors measured by the questionnaire, i.e., Psychological Stress and Social Behaviour. Analysis generally showed significantly high stress and social withdrawal for the patient group, particularly at initial consultation and after an unsuccessful treatment cycle as compared to the control group. Moreover, men whose partners eventually conceived scored significantly lower on stress than men whose partners did not conceive. Similar results have also been reported for Western infertile patients. The implications of the findings are discussed. PMID- 10840883 TI - On perceived conflicts between religion and science: the role of fundamentalism and right-wing authoritarianism. AB - Volunteers from fundamentalist churches and a Psychology of Religion class (N = 77) completed Altemeyer and Hunsberger's 1992 Fundamentalism Scale, Altemeyer's 1988 Right-wing Authoritarianism Scale, and answered questions about science, religion, and their relationship. Scores on the scales were highly positively correlated. Neither orientation correlated with seeing science as improving life, and both correlated with being troubled by newer developments in science such as organ transplants or genetic engineering. Partial correlations showed that both orientations favored religious beliefs over scientific data when there was a perceived conflict. Three subscales of right-wing authoritarianism clarified how authoritarianism correlated with other measures, thereby supporting a multidimensional conceptualization of right-wing authoritarianism. PMID- 10840884 TI - Self-efficacy and depression among Iranian college students. AB - The present study examined the correlations of depression and self-efficacy among college students. Responses of 200 college students to the Self-efficacy Scale and Middlesex Hospital Questionnaire showed that there is a negative correlation between the two sets of scores. PMID- 10840885 TI - Patients' desire to participate in decision-making in psychiatry: a questionnaire survey in Japan. AB - Japanese national sentiment has been described as paternalistic, which has potentially wide-ranging implications for the manner in which psychiatric patients should participate in medical decision-making. To examine the extent and possible determinants of the desire to participate in medical decision-making among Japanese people, we distributed a packet of questionnaires to 747 (nonmedical) university students and 114 of their parents. The questionnaires included an imaginary case vignette of psychotic depression. The participants were asked whether they would want various types of medical information, i.e., diagnosis, aetiology, treatment, outcomes, medical charts, etc., disclosed to them were they in such a psychiatric condition. Also included was the 1995 Scale for Independent and Interdependent Construal of the Self by Kiuchi. More than half of the participants wanted all the types of medical information disclosed to them. Those participants who wanted to have all types of information disclosed to them (n = 413) as compared to those who did not want to know at least one type of information (n = 445), tended to be male and to have an educational background in psychiatry (9.7% vs 5.4%) as well as an assertive attitude as indicated by a higher score on Independence on the Scale for Independence and Interdependent Construal of the Self. These results suggest that the Japanese in this sample are more likely to want to make an autonomous contribution to the psychiatric decision-making process and that less desire for information can be predicted by some demographic and personality factors. PMID- 10840886 TI - Intersibling agreement for Goldberg's big five adjective markers. AB - In a sample of 240 college students intersibling agreement was examined for Goldberg's 100 unipolar Big Five adjective markers. Participants showed self enhancement by rating themselves more favorably on three of the five traits (Agreeableness, Conscientiousness, and Culture/Intellect); however, self-ratings on Neuroticism were higher than siblings' ratings. Correlations among raters were moderate (mean r = .41) and comparable to values obtained in studies using peer ratings. The type of the sibling relationship, based on ratings of relationship quality, moderated the rank-order measures but not the mean agreement. PMID- 10840887 TI - Substance abusers: role of personal and professional role traits in caregivers' causal attributions. AB - Substance abuse continues to be a major health problem compounded by caregivers' negative attitudes toward these patients. We investigated attributions 55 primary care physicians and 315 senior medical students make toward substance abusers. Half of both groups expressed negative causal attributions, with women slightly less negative than men. Mental models based on LISREL regression coefficients showed that higher negative attributions by both physicians and students were related to their increased authoritarianism and depressed mood. Medical students choosing careers in primary care specialties, including psychiatry, expressed a less negative attributional style toward substance abusers than those students entering nonprimary careers. Health professional educators may find that using attribution theory to redefine successful outcomes in management of substance abuse can result in better attitudes for caregivers. PMID- 10840888 TI - The finesse of logotherapy reply to Kovacs (1999) AB - Kovacs (1999) raises several important points in his discussion of logotherapy. The practical dimension (applicability) of logotherapy has always been its most valuable asset. Patients on their own may recognize voids in their lives even without having any explicit knowledge of Frankl's thought. PMID- 10840889 TI - Some empirical evidence for ecological dissonance theory. AB - Using Festinger's cognitive dissonance theory as a model, the extension to Barker's ecological theory, referred to as ecological dissonance theory, was developed. Designed to examine the motivational dynamics involved when environmental systems are in conflict with each other or with cognitive systems, ecological dissonance theory yielded five propositions which were tested in 10 studies. This summary of the studies suggests operationally defined measures of ecological dissonance may correlate with workers' satisfaction with their jobs, involvement with their jobs, alienation from their work, and to a lesser extent, workers' conflict resolution behavior and communication style. PMID- 10840891 TI - Relationship of birth order and the marketing-related variable of materialism. AB - The relationship between the birth order and materialism scores was investigated using materialism conceptualized as a consumer value. Data were collected from 275 alumni of a major southwestern university. The analysis indicated that first borns in this sample scored significantly lower on materialism than younger siblings. PMID- 10840890 TI - Stability and factor structure of the Jackson Personality Inventory--Revised. AB - The Jackson Personality Inventory--Revised comprises 15 bipolar scales and five cluster scores concerning an individual's interpersonal patterns of interaction, cognitive styles, and value orientation. Recent reviews of this revised version raise questions about test-retest stability as well as the factor structure on which cluster scores are based. 74 men and 33 women (29-63 years of age, M = 42.3) completed the inventory while participating in a continuing education program. Of these 45 participated in a second session 13 wk. later. Test-retest correlations are significant, with 12 of the 15 scales having correlations > or = .75. Intercorrelations among all subscales indicate that the Jackson subscales for the most part remain distinct from each other ranging from .01 to .59. A Principal Components Analysis with a varimax rotation yielded three factors that parallel the NEO big five, i.e., Openness, Neuroticism, and Extroversion and replicated previous factor structure found for both versions of the Jackson inventory. The fourth and fifth factors here were labeled Trustworthy and Organization; however, the composition of these factors across several studies appears to be unstable, suggesting optimal certainty when interpreting the clusters of subscales associated only with Openness, Neuroticism, and Extroversion. Further research may help clarify the instability associated with the other factors of this inventory. PMID- 10840892 TI - Relationships of collective orientation and cohesion to team outcomes. AB - The relationships of collective orientation and cohesion to team outcomes were compared for 373 high school students on 12 different types of two-person teams performing simple problems in mathematics. Scores on collective orientation and cohesion were highly correlated with each other and with team outcomes but not statistically distinct from each other. PMID- 10840894 TI - The five-factor model and driving behavior: personality and involvement in vehicular accidents. AB - Participants completed both the NEO-PI-R personality measure and measures of prior involvement in driving accidents. Significant negative correlations were found between the factor of Agreeableness and the total number of driving tickets received as well as the sum of combined at-fault accidents, not-at-fault accidents, and driving tickets received by participants. Implications and potential future directions for research are discussed. PMID- 10840893 TI - An initial investigation of validation of the Matrix Analogies Test-Expanded Form in Ecuador. AB - This is a first preliminary study of the validity and reliability of the Matrix Analogies Test--Expanded Form in South America. Participants were 104 Spanish speaking children between the ages of 5 and 17 years living in Ecuador. Values of Cronbach alpha ranged from .87 to .92 for the 4 groups of items and was .95 for the total score. Raw scores on the MAT increased across ages. Scores of boys did not differ significantly from those of girls. Total test scores correlated significantly with scores on the Raven Standard Progressive Matrices (r = .62, p < .005; r = .82 before controlling for age). A principal factor analysis conducted to provide evidence of the test's construct validity indicated that all four sets of items loaded substantially on one unrotated factor, presumed to be g. In sum, these results suggest that the test is a valid and reliable nonverbal measure of general cognitive ability in this population. PMID- 10840895 TI - Note on matrices of random numbers. AB - The statistical and structural characteristics of 13 matrices of random numbers in which both the cells and the entries were randomly chosen are discussed. Each matrix was explored considering row means, standard deviations, and correlations as well as column means, standard deviations, and correlations. A study concerning the sequential arrangement of digits was performed by finding out in tables of random numbers how many times the values 0 to 9 are followed by any other digit. Analyses indicate clear factor structures when factor analyzing correlations of rows and of columns and when examining sequential arrangements, concluding that for a given set of digits it is possible to assert both randomness and nonrandomness depending on how the data are examined. PMID- 10840897 TI - Critique of recent empirical research on insight and symptomatology in schizophrenia. AB - During the past decade there has been a resurgence of interest in investigating the relationship between insight and symptomatology among clients with schizophrenia. The breadth and depth of the articles have dramatically increased over the past 10 years, including the number of empirical research studies. This article summarizes the strengths and limitations of the empirical research focused on the association between insight and severity of psychotic symptoms and published between 1990 and 1999. PMID- 10840896 TI - Use of the Abuse Disability Questionnaire in screening a clinical outpatient sample of women. AB - A sample of 50 women who sought services at a community mental health center were administered screening questionnaires which measure impairment from abuse and mental symptomatology. Scores on Psychiatric Symptomatology and Depression were correlated with impairment associated with partner abuse. Impairment was intermediate between those of a shelter sample and well-functioning community sample. The importance of screening for impairment from partner abuse in women seeking outpatient mental health care was emphasized. PMID- 10840898 TI - Behavioral and self-concept differences in tattooed and nontattooed college students. AB - 235 college students rated themselves on a series of bipolar adjectives and answered questions about their involvement in a variety of "risky" behaviors, including tattooing and body piercing. 29 tattooed students rated themselves as more adventurous, creative, artistic, individualistic, and risky than those without tattoos. The 98 tattooed males considered themselves more attractive. Behaviorally, those with tattoos reported smoking more cigarettes. Tattooed men also reported more sexual partners, were more likely to report they had been arrested, and were more likely to have body piercings. The 21 tattooed women were more likely to report use of drugs other than alcohol, shoplifting, and body piercings in places other than their ears. PMID- 10840899 TI - Steven Stack's ecological research on suicide applied to homicide. AB - In a sample of 72 nations in 1980, birth rates proved to be the strongest predictor of national homicide rates. PMID- 10840900 TI - Preference for numerical information among Arab students. AB - To investigate the preferences for numerical information of 157 tenth grade students (M = 16.1 yr.) in the United Arab Emirates, an Arabic translation of a questionnaire of Viswanathan developed, validated, and moderately stable over time was administered. Analysis indicated the preferences of Arab students for numerical information was positive. Also, these Arab students scored higher than American students in contrast with those in the Viswanathan study but the difference was not statistically significant using an independent t test. PMID- 10840901 TI - Common prescriptions for psychology derived from dialectical materialism and chaos theory. AB - During the entire Soviet period (1917-1991), Russian psychologists labored to create a psychology which would be consonant with Marxist-Leninist assumptions derived from dialectical materialism. Some of their early prescriptions, in particular those put forward by Konstantin N. Kornilov in the 1920s and early 1930s, are identical to strategies being advanced by contemporary American psychologists who propose that chaos theory and nonlinear meta-modeling techniques in general, given advances in computer and television technologies, can be designed for research capable of dealing with the complexities, nonlinearities, self-organizational processes, and abrupt transformations characteristic of human psychological functioning. PMID- 10840902 TI - A phenomenological investigation of being bored with life. AB - The majority of research on boredom concerns the situational aspects of boredom, such as monotonous stimuli and decreased arousal; however, clinical concerns such as addictive behaviors involve people who experience boredom habitually. This paper describes a phenomenological study focused on 6 persons (16 to 67 years old) who experienced habitual boredom, those who were bored with their lives. PMID- 10840903 TI - Psychotherapy with multiple-sclerosis patients. AB - We wanted to find out if psychotherapy may influence the course of the physical aspects of multiple sclerosis and the consequences of psychotherapy for coping processes. 46 patients diagnosed with multiple sclerosis who had chosen to undergo a 1-yr. group psychotherapy treatment were compared with a control group of 24 multiple-sclerosis patients without such treatment. They were given the Giessen test (personality test), the Achievement Capacities Questionnaire by Kesselring, an intensive interview as well as the content analysis scales of verbal behavior by Gottschalk and Gleser. The various tests were carried out at each of four times of measurement with a 2-yr. follow-up. There were significant changes in the area of relationships and aggressive loosening (interview) between the Therapy and Control groups. Several changes were also found with regard to physical symptoms (Achievement Capacities Questionnaire) in the Therapy group compared to the Control group, e.g., increases in physical mobility and decreases in care of the body. The decreases appear to be a known effect of therapy with psychosomatic disorders. We interpret it psychoanalytically as resistance against releasing anxiety of counter-cathected motives which multiple sclerosis helps to keep unconscious. In a follow-up, the Therapy group showed greater optimism and physical improvements, e.g., decrease in feeling cold and lack of energy. Some positive changes appeared in both groups, such as, for example, an improvement of cognitive impairment (Gottschalk & Gleser). It appears that the attention from the research itself may have affected both groups because some members of both groups were in contact and hence the Control group was also informed about the research project and its underlying hypothesis. PMID- 10840904 TI - A biosocial interaction in predicting early onset of offending. AB - Research has shown several factors increase the likelihood of early onset of offending; however, interactions among prenatal risk factors and sociological factors in predicting early onset have been a neglected area of research. The purpose of this study was to test the interactive effect of material cigarette smoking during pregnancy and the absence of the father from the household in predicting early onset of offending. The longitudinal data utilized for this study of 215 inner-city, African-American youth offenders were taken from the Philadelphia portion of the Collaborative Perinatal Project. A logistic regression analysis indicated that the combined effect of maternal cigarette smoking and absence of father from the household had a significant influence in predicting early onset of offending beyond the direct effects of the independent variables while controlling for sex of youth. PMID- 10840905 TI - Culture and the economy. AB - In a sample of 18 industrialized nations, measures of national character did not improve the power of socioeconomic variables in predicting unemployment rates. PMID- 10840906 TI - Immigrant and Native-born Mexican-American parents' involvement in a public school: a preliminary study. AB - Mexican-American parents of Texas elementary school students were surveyed to compare the types of school involvement in which immigrant and U.S.-born parents engage. Those completing the questionnaire included 246 mothers and 39 fathers born in Mexico as well as 95 mothers and 13 fathers born in the United States. More immigrant parents than U.S.-born parents indicated they helped their children with school work, attended school board meetings, volunteered at school, participated in parent-teacher conferences, went to school functions, served as room mother, engaged in school fundraising, and were present during parent advisory committee meetings. PMID- 10840907 TI - Maternal stress during pregnancy, its objectivation by ultrasound observation of fetal intrauterine movements and child's temperament at 6 months and 6 years of age: a pilot study. AB - Niederhofer showed in 1994 that maternal stress during pregnancy seemed associated with development of personality in early childhood. This study examined the correlation between maternal stress during pregnancy for 121 women, its ultrasound objectivation by observation of intrauterine fetal movements, child's temperament in early childhood, and child's marks at the age of 6 years while controlling for possible confounding variables. Only intrauterine fetal movements were not associated either with stress during pregnancy or early personality. PMID- 10840908 TI - Measuring self-efficacy among drug-involved probationers. AB - Self-efficacy has been shown to be related to outcomes from interventions for alcohol and tobacco abuse but relatively little attention has been focused on it in evaluations of treatment for illicit drug abuse. Almost no research has examined offenders involved with drugs. The current study, therefore, adapted the Alcohol Abstinence Self-efficacy Scale of DiClemente, Carbonari, Montgomery, and Hughes and administered it to 250 probationers mandated to 6 months of residential treatment. With some modifications, confirmatory factory models replicated four previously reported dimensions, e.g., Negative Affect, Social/Positive, Physical and Other Concerns, Cravings and Urges. Findings also indicated high construct validity for the Alcohol Abstinence Self-efficacy Scale as adapted here. Studies are needed to examine the use of self-efficacy as a prospective measure of treatment progress and to explore its association with outcomes from corrections-based treatment. PMID- 10840909 TI - Preliminary development of the Body Image Affect Scale. AB - A new body-image questionnaire was developed to measure the affect associated with a negative body image. Responses showed a Cronbach coefficient alpha of .84 and a negative correlation of -.21 with using humor in times of stress. PMID- 10840910 TI - Sex differences in perceptions of romantic acts in Portuguese adolescents. AB - The study was done to examine sex differences in the importance attributed to romantic acts by adolescents. On a 15-item Romantic Acts questionnaire of Neto participants rated the importance of each act. Four factors of Offerings, Physical Intimacy, Altruistic Behavior, and Sexual Behavior were found. The 108 girls judged physical intimacy as more important than the 79 boys. PMID- 10840911 TI - Effects of deceptive packaging and product involvement on purchase intention: an elaboration likelihood model perspective. AB - From an Elaboration Likelihood Model perspective, it was hypothesized that postexposure awareness of deceptive packaging claims would have a greater negative effect on scores for purchase intention by consumers lowly involved rather than highly involved with a product (n = 40). Undergraduates who were classified as either highly or lowly (ns = 20 and 20) involved with M&Ms examined either a deceptive or non-deceptive package design for M&Ms candy and were subsequently informed of the deception employed in the packaging before finally rating their intention to purchase. As anticipated, highly deceived subjects who were low in involvement rated intention to purchase lower than their highly involved peers. Overall, the results attest to the robustness of the model and suggest that the model has implications beyond advertising effects and into packaging effects. PMID- 10840912 TI - Effects of picture and even-a-penny-will-help appeals on anonymous donations to charity. AB - The present research explored the effectiveness of a picture and the phrase "even a penny will help" on contributions to charity. Two experiments were conducted, one in the laboratory and one in the field. In both experiments the manipulation of a pleasant picture and the phrase created four types of signs: (1) picture-no phrase, (2) phrase-no picture, (3) picture-phrase, and (4) no picture-no phrase. In the field experiment patrons of local business anonymously put money in a donation box that displayed one of the four signs. In the laboratory experiment, 129 undergraduate students were randomly assigned to view one of the four signs placed on a donation box for a local charity. Analyses for both studies showed that more money was donated when the boxes displayed pictures. The phrase "even a penny will help" had no significant effect on donations. PMID- 10840913 TI - Examining the validity of the Academic Motivation Scale by comparing scale construction to self-determination theory. AB - This study examined the construct validity of the Academic Motivation Scale. Specifically, subscale correlations were examined to assess whether support for a continuum of self-determination would be provided. The three types of Intrinsic Motivation were significantly and positively correlated with each other .67, .62, and .58, while the three types of Extrinsic Motivation were significantly and positively intercorrelated .50, .49, and .45. The former subscales, however, correlated higher with Introjected Regulation than Identified Regulation, suggesting that Introjected Regulation may be indicative of more self-determined behavior than has previously been believed. Also, the Intrinsic Motivation To Accomplish subscale had a stronger relationship with two of the Extrinsic Motivation subscales, Identified Regulation and Introjected Regulation, than did the Extrinsic Motivation subscales with each other. This suggests that the differences between Extrinsic and Intrinsic Motivation are not as obvious as has been believed. Also, contrary to self-determination theory, Amotivation had a stronger negative correlation with Identified Regulation (r = -.31) than with any of the Intrinsic Motivation subscales (rs = -.27, -.19, and -.11). PMID- 10840914 TI - Odor recognition and identification: effect of labels over time. AB - The effect of labels on recognition and identification of odors over time was assessed. 30 men and 30 women were presented 20 odors; half of the participants were also told a name for the odor as a label. Five min. and 60 min. later, all participants were given 20 odors (10 from the original set, 10 new) and asked whether each odor was new or old (odor recognition). The group given labels was also asked to recall the label provided (odor identification). Analysis indicated a significant effect of time on recognition. Significantly more odors were recognized at 5 min. than 60 min. The effect of label was also significant, with recognition being better for the Label condition than the No-Label condition. As for odor identification, women identified more labels than did men. Overall, odor recognition was better with labels soon after exposure, and the women were better at remembering the labels than the men. PMID- 10840915 TI - Some late nineteenth century perspectives on sex and emotional expression. AB - A review of nineteenth century popular literature indicates a deep and sustained public interest in sex differences in emotional expression. The conclusions advanced by popular writers included a catalog of perceived sex differences, reinforced by an essentialist philosophy that provided justification for the separation of sexual spheres and restrictions on political, educational, and vocational opportunities for women. Current scientific research on sex differences appears in popular media and is often presented in the context of an essentialist philosophy comparable with that which was dominant in the nineteenth century. Unfortunately, the subtleties and complexities of sex differences are not always communicated to the public and there is thus a potential for misinterpretation or even misuse. PMID- 10840916 TI - Job satisfaction of Asian Americans. AB - Since Asian Americans have demographic and labor force characteristics more similar to Euro-Americans than African Americans, one might predict that their job satisfaction would be more like the former than the latter. And, because Asian Americans originating from different countries are heterogeneous in language, culture, and recency of immigration, one might predict that they may report obtaining different amounts of satisfaction from their jobs. However, data from 21 nationally representative opinion surveys from 1972 through 1996 suggest the opposite. Asian Americans (n = 199) reported job satisfaction more like African Americans (n = 1,231) than Euro-Americans (n = 10,709), and Asian Americans from China (n = 53), Japan (n = 44), India (n = 55), and the Philippines (n = 47) reported similar job satisfaction. These differences persisted when age, education, occupation, and personal income were held constant. PMID- 10840917 TI - Age and sex differences in paranormal beliefs: a response to Vitulli, Tipton, and Rowe (1999) AB - Vitulli, Tipton, and Rowe (1999) report evidence of age and sex differences in the strength of paranormal beliefs. An alternative interpretation of their data is offered in terms of differential item functioning. It is suggested that respondents' interpretation of paranormal belief test items may vary with age and sex, and that such differences in the strength with which such beliefs are endorsed has not been conclusively established by Vitulli, et al. PMID- 10840918 TI - On the nature and properties of appeals used in direct-to-consumer advertising of prescription drugs. AB - The past decade has seen a steady rise in expenditures for direct-to-consumer pharmaceutical advertising. While total revenues across all media are approaching the $1 billion dollar mark, surprisingly little is known about the effectiveness of these types of advertisements, including the appropriateness of various forms of emotional and informational appeal. A content analysis of direct-to-consumer advertising in 24 popular magazines shows that these advertisements are found in every category of magazine, the advertisements employ a mix of informational and emotional appeals, all types of emotional appeals are used, and to date, the type of appeal (emotional and/or informational) tends not to be based on the type of drug advertised. Implications of this content analysis are considered and directions for research on appeals used in direct-to-consumer advertising are suggested. PMID- 10840919 TI - Organizational change: initial results in a health care setting. AB - An applied study was conducted in a federal health care facility to assess change in organizational climate variables over a 7-yr. period. Using the Survey of Organizations--2000, preliminary results show improvements on five climate indicators from Time 1 to Time 2 of the survey. Potential explanations for the positive changes are provided. PMID- 10840920 TI - Comparing leaders' ratings to targets' self-reported resistance to task assignments: an extension of Chester Barnard's zones of indifference. AB - This description and preliminary test of target resistance compared leaders' with targets' ratings of the targets' willingness to perform the nature of tasks assigned by leaders. Barbuto's concentric zones were used to test this difference in perceived resistance. The concentric zones--Preference, Indifference, Legitimate, Influence, and Noninfluence--were examined from both leaders' and targets' perspectives. Response from 201 leaders and targets demonstrated that leaders and followers generally reported the anticipated resistance of assigned tasks differently. Specifically, leaders rated the tasks they assigned to targets as less enjoyable, undesirable, more above the call of duty, and more likely to be resisted than targets self-reported. PMID- 10840921 TI - Students' perceptions and reported use of their memories after a memory improvement course. AB - 23 college students who had taken a memory-improvement course completed a questionnaire at three different times: the beginning of the course, the end of the course, and two months after the end of the course. The questionnaire assessed the students' perceptions of their memory abilities, e.g., how good their memories were and how confident they were in their memories and knowledge about their memories, and their reported use of the memory techniques; it did not assess actual memory performance or ability. The students' responses to the questionnaire were compared with those of 23 control students who completed the questionnaire at the same three times. The memory students reported more positive changes in how they perceived their memory abilities and knowledge, and these reported changes in perceptions were still evident two months later. The students also reported that they were still using their memory techniques two months later. PMID- 10840922 TI - Loneliness and the life cycle. AB - Loneliness is a prevailing experience which every person has experienced. This subjective experience is influenced by one's personality and situational variables. In the present study, the influence of age and sex on the experience of loneliness were examined. 711 participants volunteered to answer an 82-item yes/no questionnaire on their loneliness experience and its meaning. Four age groups were compared: 106 youths (13-18 years old), 255 young adults (19-30 years old), 314 adults (31-58 years old), and 36 seniors (60-80 years old). Within and between sex comparisons indicated that loneliness is indeed affected by one's age and sex. PMID- 10840923 TI - Materialism and credit card use by college students. AB - Much has been written in the popular press on credit card use and spending patterns of American college students. The proliferation of credit cards and their ease of acquisition ensure that students today have more opportunities for making more credit purchases than any other generation of college students. Little is known about the relationship between students' attitudes towards materialism and their use of credit cards. A study was conducted at three college campuses in the northeastern part of the United States where a total of 1,022 students were surveyed. Students' attitudes toward use of credit and their credit card balances were evaluated relative to their scores on Richins and Dawson's Materialism Scale (1992). Our findings suggest no significant difference between those individuals scoring high versus low on the Materialism Scale in terms of the number of credit cards owned and the average balance owed. Individuals high on materialism, however, significantly differed in terms of their uses for credit cards and their general attitude toward their use. PMID- 10840924 TI - Clients' perspectives on problems many years after traumatic brain injury. AB - This study investigated the types of self-reported main problems that persons report many years following a traumatic brain injury. This preliminary study is part of a large ongoing study of disability and handicap in adults following traumatic brain injury. As part of an extensive interview, subjects were asked an open-ended question regarding their current main problems which they thought resulted from their traumatic brain injury. Responses were obtained from 166 adult subjects (119 men and 47 women) whose time postinjury ranged from 9 to 24 years. Categories for responses were subsequently developed. The most commonly reported categories of problems were those relating to movement (39%), cognition (36%), and sensory impairment (31%). Findings suggest that subjects' long-term concerns were related to specific impairments more than to psychosocial, emotional, or behavioral issues. Methodological issues concerning this research are discussed in relation to findings. PMID- 10840925 TI - Variations in subject pool as a function of earlier or later participation. AB - Data were obtained from 113 participants in a university subject pool during a 16 wk. semester. Without knowing the purpose of the study, participants self selected to participate earlier (Weeks 3 and 4: n = 63) or later (Weeks 15 or 16) n = 50). Variations in scores on the NEO Personality Inventory--Revised, the Crowne-Marlowe Social Desirability Scale, the General Expectancy of Success Scale, the Shipley Institute of Living Scale, self-reported SATs and GPAs, and a measure of academic self-efficacy as a function of earlier or later participation were examined. Multivariate analysis of variance indicated that early participants differed significantly from later participants but not in predicted ways. Earlier participants scored higher on NEO PI-R Neuroticism; specifically men (n = 15) and women (n = 48) scored higher on Hostility, and women scored higher on Depression and Self-consciousness. An additional significant difference occurred for self-reported SAT Verbal scores for men, which were significantly higher for later participants. These temporal variations may represent confounds in research using university subject pools. PMID- 10840926 TI - Flight and abduction in witchcraft and UFO lore. AB - The lore surrounding the mythical Witches' Sabbat and contemporary reports of UFO abductions share three main characteristics: the use of masks, the appearance of "Men in Black," and references to flight and abduction. We review these three commonalities with particular focus on the aspect of flight and abduction. We argue that narratives of the Witches' Sabbat and UFO abductions share the same basic structure, common symbolism, and serve the same psychological needs of providing a coherent explanation for anomalous (ambiguous) experiences while simultaneously giving the experient a sense of freedom, release, and escape from the self. This pattern of similarities suggests the possibility that UFO abductions are a modern version of tales of flight to the Sabbat. PMID- 10840927 TI - Beliefs and practices of therapists who practice sexual reorientation psychotherapy. AB - There is currently controversy regarding whether sexual reorientation or conversion therapies are ethical and effective forms of treatment for dissatisfied homosexually oriented people. We present the results of a survey of 206 psychotherapists who practice sexual conversion therapy. 187 therapists said they believed homosexuality is a developmental disorder and that the 1973 decision by the American Psychiatric Association to "depathologize" homosexuality was politically motivated and unscientific. The therapists believe that the majority of dissatisfied homosexually oriented clients who seek conversion therapy benefit from it, experiencing both changes in their sexual orientation and improved psychological functioning. We conclude that therapists who persist in providing reorientation therapy do so because they believe it is an effective and ethical treatment option for their clients. PMID- 10840928 TI - A 25-year thymoma treatment review. AB - Most thymomas are stage I or II at presentation, and they have a good prognosis with surgical treatment. Higher stage thymomas are less common and their treatment is more problematic. Our center tends to attract patients with higher stage thymomas for treatment. We reviewed our experience and contrasted it with other published series. A 25-year retrospective record review of thymomas was done. 38 patients were treated. Median age was 49 years. Four had myasthenia gravis. Masaoka staging was: stage I--9; stage II--6; stage III--15; stage IVa- 4; stage IVb--4. Resection was done in 25 patients (21 had R0 resection), chemotherapy was given to 15 patients, and 27 patients received radiotherapy. Overall median survival was 55 months. Overall 5 and 10-year survivals were 30% and 18%. 5-year survival by stage was: stage I--75%; stage II--50%; stage III/IV- 23%. Negative prognostic factors on univariate analysis included presence of symptoms at presentation (p = 0.02), unresectable tumor (p = 0.06), stage III/IV (p = 0.04), and disease recurrence after resection (p = 0.0001). On multivariate analysis, only stage (p = 0.04) and recurrence (p = 0.0001) were independent predictors of survival. All patients who recurred after resection eventually died of disease. Our overall treatment results are disappointing, but we had higher stage patients than reported by most other centers. Early stage thymomas are suitable for complete surgical resection, and the prognosis is favorable. However, higher stage thymomas (stage III and higher) pose problems for complete surgical resection and their prognosis is poor. Newer multimodality treatment approaches are indicated for higher stage thymomas. PMID- 10840929 TI - Tumors of the parathyroid glands. Changes in clinical features and in noninvasive localization studies sensitivity. AB - Primary hyperparathyroidism is the most common cause of hypercalcemia and 80-85% of the patients have parathyroid tumors. The purpose of this retrospective review was to analyse whether differences exist between patients with parathyroid tumors treated in the 1980s and 1990s. Between 1980-1997, 253 patients underwent initial surgical neck exploration for hyperfunctioning parathyroid tumors. Renal (polyuria, nocturia, renal colic due to lithiasis), rheumatologic (bone and joint pain), neurological (fatigue, memory loss, depression) and gastrointestinal (dyspepsia, anorexia, nausea) symptoms were recorded and main biochemical parameters were measured. In all patients one or more preoperative localization procedures were carried out prior to successful parathyroidectomy, and the confirmation of imaging findings was obtained after surgery. The patients were divided in two groups. Group A: 121 (47.8%) patients who underwent surgery from 1980-1989; Group B: 132 (52.2%) patients in whom parathyroidectomy was performed from 1990-1997. There were no differences (p=NS) between the two groups in average age, preoperative serum creatinine and intact-PTH levels. Symptoms were most common in Group A, and pre-operative serum calcium levels were significantly lower in Group B. Ultrasonography (n=191) sensitivity did not improve significantly (82.8% vs 82.9%), but positive predictive value (PPV) was higher (89.8% vs 96.0%). CT-scan (n=73) sensitivity was 79.2% and 82.6%, and PPV was 95.0% and 100% in Groups A and B, respectively. 201Tl/99mTc subtraction scintigraphy (n=111, Group A) was 84.6% sensitive (PPV=92.6%) whereas 99mTc sestamibi scanning (n=90, Group B) was 85.1% sensitive (PPV=96.1%). In conclusion, the clinical features of parathyroid tumors has changed in the nineties and increasing asymptomatic pHPT rate has been found. Although sensitivity and PPV of preoperative localization procedures has improved moderately, at present noninvasive techniques may offer excellent results and should be used in all patients with suspected parathyroid tumors. PMID- 10840930 TI - Cisplatin, epirubicin and cyclophosphamide (PEC) in the treatment of advanced ovarian cancer. AB - We report the long-term results of a series of patients affected by advanced epithelial ovarian cancer treated with the PEC combination (cisplatin 60 mg/m2, epirubicin 60 mg/m2 and cyclophosphamide 750 mg/m2, all at day 1, every 21 days). Response was evaluated after three cycles, and treatment continued in responsive patients. A total of 80 patients with a median follow-up of 55 months were studied. Fifty-eight patients with stage III ovarian cancer and 22 patients with stage IV received PEC as primary treatment (41 patients), or for residual disease after surgery (37 patients), or for relapsed disease after primary surgery (2 patients). The overall response rate was 67.5% (20.0% complete response, 47.5% partial response), with 22.5% stable disease and 3.7% progressive disease. Median progression free survival was 13.0 months, and median survival was 25 months. Grade III-IV toxicity was moderate: leukopenia 20.0% of patients, thrombocytopenia 5.0%, anemia 16.2%. No cardiac toxicity was observed. In conclusion, the PEC combination, an anthracycline-containing platinum-based regimen, proved to be effective in advanced ovarian cancer, in terms of response rate and overall survival. The regimen was devoid of significant toxicity and in particular of cardiac toxicity. PMID- 10840931 TI - Letrozole for the treatment of pretreated advanced breast cancer patients: preliminary report. AB - Twenty patients (pts) with metastatic breast cancer with disease progression, previously treated with chemotherapy and tamoxifen, were administered oral letrozole (2.5 mg/day) therapy. Fifteen of the patients were postmenopausal and 5 were premenopausal. Ten were estrogen receptor (ER)-positive, 7 were unknown and 3 were ER-negative. All the patients were assessed after 6 months (mo) of chemotherapy. Nine pts (45%) presented a partial response (PR), five (25%) had a stable disease (SD) and six (30%) had a progressive disease (PD). In the pts with PD, six out of 15 (33%) obtained a PR while undergoing tamoxifen therapy. The treatment caused no significant toxicity. PMID- 10840932 TI - Dacarbazine-based chemotherapy for metastatic melanoma: thirty-year experience overview. AB - Dacarbazine (DTIC) is the only single-agent approved by the Food and Drug Administration for treating metastatic melanoma. With DTIC as single agent, an approximately 20% objective response rate can be achieved with median response duration of 5 to 6 months and complete response rates of 5%. Current status of DTIC single agent and DTIC-based combination chemotherapy has been extensively reviewed in this article. Moreover, future directions including new combination chemotherapies and/or new therapeutical approaches have been considered. The addition to DTIC of agents such as cisplatin, nitrosoureas and tubular toxins has been reported to yield high response rates, up to 40%, in single-institution phase II trials. Historically, promising combination regimens like BOLD (bleomycin, vincristine, lomustine and DTIC) and CVD (cisplatin, vinblastine and DTIC) have induced responses on metastatic lesions to the liver, bone and brain, commonly unresponsive to DTIC alone, even though have failed to produce impact on patient survival. Several other studies have suggested a significant enhancement of antitumor effect associated with the addition of tamoxifen to various cytotoxic regimens. The four-drug combination CBDT (cisplatin, carmustine, DTIC and tamoxifen) or "Dartmouth regimen" has yielded high response rates, up to 55%, with continuous, maintained, complete responses, up to 82 months, in a subset of patients, that is considerably longer than observed with other combinations. Some authors recommend CBDT as reference therapy, even though recently presented results of a randomized phase III trial of CBDT versus DTIC alone, show no statistical difference in survival between the two groups. While a survival benefit from DTIC-based chemotherapy or DTIC alone has never been shown in metastatic melanoma patients and, therefore, the survival has remained unchanged over the past 30 years, some long term survivors have been reported with the "Dartmouth regimen" and/or with high dose interleukin-2 (IL-2) based regimens whose role is going to be defined in prospective randomized phase III trials. On the other hand, the better understanding of the mechanisms responsible for melanoma chemoresistance and the development of new therapeutical strategies could change the scenario in the next future. PMID- 10840933 TI - Nerve-sparing surgery in rectal cancer: feasibility and functional results. AB - Traditional rectal cancer surgery has been burdened with a high rate of sexual and urinary dysfunctions due to intraoperative injury or the cutting of the sympathetic and/or parasympathetic nerves. The experience acquired in the last ten years with total mesorectal excisions has permitted the use of the "nerve sparing" technique. The present study regards 239 patients from two surgical centres, most of whom underwent sphincter-saving radical surgery between 1994 and 1998 with the above mentioned technique for resectable colon cancer. Details regarding the technique were recorded in the last 58 patients, in order to examine the severity of the surgical damage. The subgroup with the longest follow up, which included 36 patients, was diagnostically evaluated by a surgeon, psychologist, urologist and neurologist to analyze the risk of sexual and urinary dysfunctions. A complete nerve-sparing was performed in 86.3% of the cases. The parasympathetic nerve trunks were those most often damaged because of perineural tumor spreading. Partial to complete sexual impotence was observed in 44% of the patients and surprisingly, preoperative dysfunctions were detected by means of the multidisciplinary approach in one third of these patients. Therefore, only 30.5% of the patients presented with strictly postoperative sexual impotency, above all, those who had undergone high-dose preoperative chemoradiation for T3 or T4 middle to low rectal cancer. A prospective study was initiated to evaluate the genitourinary dysfunctions after rectal cancer surgery in all of the clinical phases by means of a multidisciplinary approach aimed at functional recovery and improved quality of life. PMID- 10840934 TI - Surgical approach to non-small cell lung cancer involving the chest wall. AB - Treatment of NSCLC invading the chest wall (T3) remains controversial. Surgery is undoubtedly the only chance for these neoplasms, but its role regarding the T3N2 tumors is highly questionable. Between 1975 and 1994, 710 NSCLC patients underwent surgery in our department. Of these, 38 with tumor involvement of the chest wall underwent curative resection: en bloc resection or extrapleural resection, and 31 of these patients (19 with T3N0 tumors and 12 with T3N1-N2 tumors) were available for estimating long-term survival. The overall survival was 20.5% at 5 years and 15.4% at 10 years. Patients without lymph-node involvement had a survival rate of 26.2% at 5 years and 19.27% at 10 years. No patient with T3N2 tumor was alive 5 years after surgery. Patients with T3N1 tumor had a survival rate of 16.7% at both 5 and 10 years. The difference between T3N0 and T3N2 tumors was statistically significant. Neither histologic type nor depth of chest wall involvement had a significant impact on survival. En bloc or extrapleural resection, if curative, can be effective in T3N0-N1 tumors. Surgery is inadequate for the treatment of T3N2 tumors with chest wall involvement. PMID- 10840935 TI - Suppression of N-nitrosomethylbenzylamine-induced rat esophageal tumorigenesis by dietary feeding of auraptene. AB - The modifying effects of auraptene on N-nitrosomethylbenzylamine (NMBA)-induced esophageal tumorigenesis were investigated in male F344 rats. At 5 weeks of age, all animals, except those with the test chemical alone and control rats, received s.c. injections of NMBA (0.5 mg/kg body weight/injection, three times per week) for 5 weeks. At the end of the study (20 weeks), 75% of the rats treated with NMBA alone had esophageal neoplasms (papillomas). However, the groups who received a dose of 500 ppm auraptene during the initiation phase developed significantly reduced incidence of tumors (39%; P<0.05). Exposure to auraptene (500 ppm) during the post-initiation phase also decreased the frequency of the tumors (29%; P<0.01). The reduction of the incidence of severe dysplasia was obtained when auraptene was administered in the post-initiation phase (P<0.05). Cell proliferation in the esophageal epithelium determined by proliferating cell nuclear antigen (PCNA) was lowered by auraptene (P<0.01). Blood polyamine contents in rats who received NMBA and the test compound were also smaller than those of rats that received the carcinogen (P<0.05). These findings suggest that dietary auraptene is effective in inhibiting the development of esophageal tumors by NMBA when given during the initiation as well as post-initiation phases, and such inhibition is related to suppression of cell proliferation in the esophageal epithelium. PMID- 10840936 TI - Decline of met-enkephalins concentration after interleukin-2 subcutaneous administration due to renal carcinoma. AB - The authors have described the connections between nervous and immune systems. Endogenous opioids are one of the factors linking both systems. Endogenous and exogenous opioids can modify the function of interferons, humoral factors, antibody production and lymphocytes' activation. Concurrently, some cytokines can modify the endogenous opioid system. The aim of this study was to asses whether the subcutaneous administration of IL-2 influences the Met-Enkephalins concentration in serum after IL-2 single administration due to renal cancer. The Met-Enk level was estimated with RIA method. The IL-2 single administration results in a significant decrease of Met-Enk in serum. PMID- 10840937 TI - Leukemia-associated gene rearrangements in blood mononuclears of subjects in long terms after radiation exposure. AB - The results of electron microscopy and molecular genetic study of blood mononuclears of 220 clean-up workers after 7-10 years since Chernobyl accident are presented. An increase of lymphocytes with altered ultrastructure of nuclei and membrane has been observed. Structural polymorphism of leukemia associated bcr and rRNA genes has been analyzed using Southern blot hybridization. Allelic polymorphism of bcr gene with allele distribution characteristic of myeloid leukemia and rearrangements of rRNA genes have been revealed in 11,5% of clean-up workers under study. PMID- 10840938 TI - The expression of products of oncogens c-erbB2 and EGFR and proliferating antigens Ki67 and PCNA in primary invasive ductal cancer of female breast. AB - The aim of the study was to examine the consecutive series of primary ductal invasive tumours and find out: a) the expression of some biological cellular parameters as proliferating antigens Ki67 and PCNA and products of gene EGFR, erbB2; b) correlation between the levels of expression of those factors and classical prognostic factors, such as diameter of tumour, status of axillary lymph nodes, status of steroids receptors, the degree of histological differentiation. We found that: 1. The presence of the expression of oncoproteins c-erbB2 and EGFR, high index IP PCNA, Ki67 and low levels of steroids receptors correlates with high histological malignancy (Bloom III0); 2. The lack of expression of oncoproteins c-erbB2, EGFR and low index IP PCNA, Ki67 correlates with high levels of steroids receptors; 3. The estimation of high levels of index IP PCNA, IP Ki67 can be helpful for separate tumours of high proliferating activity; 4. The expression of oncoprotein c-erbB2 and EGFR does not correlate with the diameter of tumour as well as with the involvement of axillary lymph nodes. It seems that the estimation of proliferating antigens together with the expression of oncoproteins might have greater prognostic value than the estimation of one of these factors. PMID- 10840939 TI - Prognostic factors on 45 cases of phyllodes tumors. AB - The authors, in order to obtain a diagnostic index for phyllodes tumors and identified histological parameters that will predict the clinical course of this neoplasm, developed a histological degree of aggressiveness based on specific histological parameters, including: stromal:gland ratio, tumor margins, mitotic index and degree of stromal pleomorphism. Three categories were established: benign, intermediate and malignant. The probability of recurrence was estimated by the relative risk and by a multivariate Cox analysis. A strong and significant association was observed between this histological index and recurrence. The relative risk was 6.0 for intermediate lesions and 11.4 for malignant lesions when compared with the benign category. The microscopic examination of all axillary lymph nodes was negative for metastatic disease. In the multivariate analysis, the stroma:gland ratio was the strongest predictor for recurrence. These results indicate that by assigning a numerical value to certain histopathologic variables a better correlation with the clinical outcome of the patient can be obtained. PMID- 10840940 TI - Enzyme/prodrug gene therapy for human colon cancer cells using adenovirus mediated transfer of the Escherichia coli cytosine deaminase gene driven by a CAG promoter associated with 5-fluorocytosine administration. AB - Escherichia coli cytosine deaminase (CD), which is a prokaryotic enzyme, converts nontoxic prodrug 5-fluorocytosine (5-FC) into the toxic chemotherapeutic agent 5 fluorouracil (5-FU). To investigate an enzyme/prodrug gene therapy for colorectal cancer, using adenoviral gene transfer of the E. coli CD gene associated with administration of 5-FC, we constructed replication-defective adenovirus vectors expressing the E. coli CD gene or lacZ gene driven by a CAG promoter (composed of a cytomegalovirus immediate early enhancer and a chicken beta-actin promotor). The present study demonstrated that an adenoviral gene transfer system using a CAG promoter induced sufficient gene expression of CD to confer the cytotoxicity of 5-FC to HT29 human colon cancer cells by converting it into 5-FU even at an moi of one. Furthermore, experimental gene therapy using intratumoral injection of the CD-expressing adenovirus with systemical administration of 5'-FC successfully suppressed the growth of established HT29 subcutaneous tumors in nude mice. These results suggest that enzyme/prodrug gene therapy using the adenoviral gene transfer of the E. coli CD gene with concomitant administration of 5-FC may be an effective strategy in the local control of colorectal cancer. PMID- 10840941 TI - Effect of retinoic acid on integrin receptors of B16F10 melanoma cells. AB - The intriguing problem of metastasis requires the spreading of metastatic cells through the basement membrane barrier. The interaction of the basement membrane with the metastatic cell is a cell surface activity involving the function of integrin receptors. Integrins are a group of alpha,beta heterodimeric proteins responsible for transducing intracellular signals on binding to the extracellular matrix proteins present in the basement membrane. To understand the role of integrin receptors in tumor metastasis, the cell surface receptor functions were modulated by All Trans Retinoic Acid (ATRA) treatment in B16F10 tumor cells. Our experimental results clearly indicate that All Trans Retinoic Acid (ATRA) inhibit metastatic potential of highly metastatic B16F10 melanoma cells by 1) downregulating the cell surface integrin receptors against ECM proteins specially laminin and vitronectin and 2) by inhibiting the 72 kd collagenase activity. PMID- 10840942 TI - Malachite green induced malignant transformation of Syrian hamster embryo (SHE) cells in primary culture: transformation is associated with enhanced expression of altered p53, bcl-2 and decreased sensitivity to apoptosis. AB - Malachite green (MG) consisting of green crystals with a metallic lustre, is highly soluble in water, cytotoxic to various mammalian cells and also acts as a liver tumor promoter. In view of its industrial importance and possible exposure to human beings, MG poses a potential environmental health hazard. We have previously reported the malignant transformation of Syrian hamster embryo (SHE) cells by MG. In this study, we have made an attempt to study the effects of MG on the induction of apoptosis in normal and MG transformed Syrian hamster embryo cells and the expression of altered p53 and bcl-2 immunohistochemically. Induction of apoptosis was detected by flow cytometry on the basis of G0/G1 hypodiploid peak, Tunel assay and DNA ladder pattern. Flow cytometric analysis showed a dose and time dependent induction of apoptosis by MG in control cells whereas induction of apoptosis by MG was marginal in transformed cells. Tunel assay and DNA ladder pattern also showed decreased apoptosis in transformed cells by MG compared to controls. Immunostaining studies showed intense staining for p53 in transformed cells whereas no staining was observed in control cells. Also transformed cells showed overexpression of bcl-2 with exclusive nuclear localization compared to controls which did not show staining. The present study indicated that MG transformed Syrian hamster embryo cells have decreased sensitivity to apoptosis compared to normal cells and overexpression of altered p53 and bcl-2 seems to be conferring resistance to MG induced apoptosis. PMID- 10840943 TI - Fibronectin and malignant disease-associated DNA-binding protein 2 in hepatocarcinogenesis in rats. AB - Changes in the plasma concentration of malignant disease-associated DNA-binding protein 2 (MAD2) and in the distribution of fibronectin and MAD2 in liver tissue were studied in Fisher-344 rats with diethylnitrosamine-induced hepatocarcinogenesis. The concentration of plasma MAD2 significantly increased as pre-cancerous lesions developed into hepatocellular carcinoma. We believe that the increased plasma concentration of MAD2 is caused by an increase in the degradation of fibronectin within hepatocellular carcinoma tumors. Therefore MAD2 may be a useful marker for the early detection of hepatocellular carcinoma. PMID- 10840944 TI - Molecular analysis of a candidate metastasis-associated gene, MTA1: possible interaction with histone deacetylase 1. AB - We previously identified a novel rat candidate metastasis-associated gene, mta1, based on its differential expression in highly metastatic cells compared to nonmetastatic cells. Furthermore, we showed that overexpression of its human counterpart, MTA1, correlated with the invasiveness or lymph node metastasis of gastric, colorectal and esophageal carcinomas. The aim of this study was to analyze the domains of the MTA1 and investigate the function(s) of this protein. Structural analysis revealed that the MTA1 protein contained a GATA-like zinc finger domain, a leucine zipper domain, a SANT domain similar to the DNA binding domain of myb-related proteins, a src homology 3-binding domain important in protein-protein interactions, two highly acidic regions characteristic of the acidic activation domains of many transcription factors, and nuclear localization signals. Immunofluorescence staining of COS-7 cells transfected with a myc epitope-tagged MTA1 expression vector clearly showed nuclear localization of MTA1. Coimmunoprecipitation of myc-tagged MTA1 and FLAG-tagged histone deacetylase 1 (HDAC1), followed by western blot analysis using anti-myc and anti FLAG monoclonal antibodies showed that MTA1 physically bound with HDAC1 in COS-7 cells. Together with the recent finding that the NURD (nucleosome remodeling and histone deacetylase activities) complex contains an MTA1-related gene product, named MTA2, MTA1 may be another component of this complex and be involved in the alteration of chromatin structure and transcription repression. PMID- 10840946 TI - A case against elective nephron sparing surgery for renal cell carcinoma. AB - A 62-year old man who incidentally was discovered to have a mass lesion in the lower pole of the left kidney underwent left radical nephrectomy. The mass was 32 millimetres in its largest diameter. Although the left adrenal gland was radiologically and macroscopically normal, histopathological examination revealed microscopic foci of metastasis. The present case urges us to be more cautious in offering partial nephrectomy for left sided tumours even though it is incidentally detected and small. PMID- 10840945 TI - Establishment and characterization of a human gastric carcinoma cell line that is highly metastatic to lymph nodes. AB - The actual mechanisms by which carcinoma cells metastasize to lymph nodes are still unclear, and there is a need to establish in vivo experimental models suitable for the investigation of lymph node metastasis. For the purpose, we established a highly lymph node-metastasizing line, designated AZL5G, derived from a human gastric cancer cell line, AZ521, which had low capacity for lymph node metastasis. AZL5G cells transplanted orthotopically in the nude mouse stomach metastasize predominantly to the regional lymph nodes, showing little potential for hematogenous metastasis. AZL5G tumors developing in the stomach and regional lymph nodes showed poorly differentiated adenocarcinoma with medullary growth, and their histologic appearance strongly resembled that of parental AZ521. The growth activities in vitro of low-metastatic AZ521 and high-metastatic AZL5G were almost the same, but the tumorigenicity in vivo of AZL5G was significantly higher than that of AZ521. AZL5G cells also showed clearly higher abilities of cell locomotion and adhesion to type IV collagen and fibronectin in vitro as compared with AZ521 cells. Flow cytometric analysis demonstrated that the expression of integrin beta1 subfamily except for alpha6 integrin was generally increased in AZL5G cells than in AZ521 cells. Especially, the expression of alpha1 and alpha2 integrins in AZL5G cells was clearly higher than in AZ521, while alpha(v)beta3 integrin, E-cadherin, ICAM-1 and CD44H were not expressed by either cell line. The cell adhesion blocking assay showed that DGEA containing peptide, which is composed of alpha2 integrin recognition sequence, significantly reduced the adhesiveness of AZL5G cells to type IV collagen as well as to type I collagen and laminin. Furthermore, the administration of anti-alpha2 integrin mAb or DGEA peptide in AZL5G-transplanted nude mice produced a significant reduction in the number of lymph node metastases. These data suggest that the up-regulation of alpha2 integrin expression by gastric cancer cells may play a critical role in the process of lymph node metastasis through the increased adhesiveness to type IV collagen. In conclusion, we established a gastric cancer cell line, AZL5G, with a highly metastatic potential to lymph nodes. This well-characterized line and its in vivo experimental model should be useful for investigation of the mechanisms of lymph node metastasis and for establishment of a new therapeutic approach for human gastric cancer. PMID- 10840948 TI - The electrophilic amination of carbanions: an unconventional new entry to C-N bond formation AB - The electrophilic amination of carbanions allows the preparation of a wide range of amines through an unconventional C-N bond-forming reaction. The concepts behind the varied synthetic approaches, classified by the nature of the aminating agent and of the organometallic species, are discussed. The mild operational conditions, the high selectivity, and the availability of the starting materials are good assets of these processes which nicely complement each other. New appealing and flexible routes can be devised, leading in several cases to the synthesis of otherwise not easily accessible N-containing compounds. PMID- 10840947 TI - Skin lesion's treatment with alfa interferon in a patient with B-CLL. AB - Treatment with recombinant alfa-2b-interferon in a patient with advanced B-CLL resulting in complete remission of skin lesions after one month therapy is reported in this paper. Interferon was administered subcutaneously three times weekly. Six months later while the treatment continued, the disease remained stable. PMID- 10840949 TI - Chiral photochemistry within zeolites AB - Chiral induction of chemical reactions continues to be one of the main concerns of chemists. While basic rules of chiral induction of thermal reactions have been reasonably established, the same is not true of photochemical reactions. Short excited state lifetime and low activation energies for reactions in the excited state(s) leave very little room for manipulating the diastereomeric transition states. Yet impressive chiral induction of photochemical reactions in the solid state has been achieved. On the other hand, chiral induction of photoreactions of organic molecules in solution continues to be inefficient at ambient conditions. We are exploring the possibility of employing zeolites as a media for achieving chiral induction during photoreactions. The motivating force for such an attempt is the fact that chiral chemistry in the solid state is not completely general due to the fact that not all molecules crystallize. To achieve chiral induction one needs a chiral perturber. Zeolites are not chiral and therefore the perturber is added to the medium. Thus the medium for a photoreaction is a chirally modified zeolite. Of the several reactions investigated, results on photoelectrocylization of tropolone alkyl ethers are discussed at length. The confined space offered by the zeolite supercage forces a reactant and the chiral inductor to interact intimately to yield enantiomerically enriched product. Due to the transitory nature of the reaction cavity in solution such close interactions are less likely in isotropic solvent media. The examples discussed herein show negligible chiral induction in solution, whereas in a zeolite one obtains induction as high as 90%. PMID- 10840950 TI - Tuning of photo- and electroluminescence of new soluble, PPV-analogous short chain compounds with naphthalene moieties AB - A series of oligomers analogous to poly(p-phenylenevinylene) (PPV), combining naphthalene and benzene as aromatic units, have been synthesized by a Knoevenagel reaction. By measuring UV/Vis spectra, photoluminescence (PL) and electroluminescence (EL), we studied the influence of the position of the naphthalene unit as well as the steric and electronic influences of cyano and alkyloxy substituents on the luminescent properties of these compounds. PMID- 10840951 TI - Diastereo- and enantioselective total synthesis of stigmatellin A. AB - Stigmatellin A (1) isolated from the myxobacterium Stigmatella aurantiaca is a powerful inhibitor of electron transport in mitochondria and chloroplasts. The first highly diastereo- and enantioselective total synthesis of this important natural product is described. Key steps in the synthesis are the alkylation of the SAMP-hydrazone (S)-13, a titanium mediated syn-diastereoselective aldol reaction, the anti-diastereoselective triacetoxyborohydride reduction of the aldol adduct (R,R,S)-16, formation of the chromone system via Baker Venkataraman rearrangement and exclusive (E) C=C double bond formation via Horner-Wadsworth Emmons reaction. PMID- 10840952 TI - Anions derived from squaric acid form interionic pi-stack and layered, hydrogen bonded superstructures with organometallic sandwich cations: the magnetic behaviour of crystalline AB - Depending on the stoichiometric ratio, squaric acid (3,4-dihydroxy-3-cyclobutene 1,2-dione, H2SQA) reacts with [(eta6-C6H6)2Cr] in THF to form the crystalline material [(eta6-C6H6)2Cr][HSQA] (1) and in water to yield [[(eta6 C6H6)2Cr]2][SQA] x 6H2O (3); it also reacts with [(eta5-C5H5)2Co][OH] in water to form [[(eta5-C5H5)2Co]2][SQA] x 6H2O (4). Compound 1 is almost isostructural with the previously reported salt [(eta5-C5H5)2Co][HSQA] (2); its structure is based on pi-pi stacks between the benzene ligands and the hydrogen squarate anionic chains (pi-pi distance 3.375 A). Compounds 3 and 4 are isomorphous and have a structure in which layers of organometallic cations intercalate with layers of water molecules hydrogen bonded to squarate dianions. All crystals contain charge assisted C-Hdelta+...Odelta- hydrogen bonds between the organometallic and the organic components, while negative O-H(-)...O(-) and O-H...O(2-) interactions are present in the pairs 1/3 and 2/4, respectively. In constrast to most organic salts of [(eta6-C6H6)2Cr]+ and [(eta5-C5H5)2Co]+ which are yellow, crystals of compounds 1-4 are orange. Reflectance spectra measured on the crystalline material 1 show the presence of an intense tail that can be assigned to a charge transfer transition through the [(eta6-C6H6)2Cr]+/[HSQA]- pi-stacking interactions, while the pi stacking in 2 causes only a broadening of the band. The magnetic behaviour of 1 and 3 has been investigated by SQUID magnetometry. Both compounds are characterised by a weak antiferromagnetic interaction between the S=1/2 Cr centres of the [(eta6-C6H6)2Cr]+ cations, which is significantly stronger in 1 due to the pi-stacking with the HSQA- anions. PMID- 10840953 TI - A novel series of copolymers containing 2,5-dicyano-1,4-phenylene-vinylene- synthetic tuning of the HOMO and LUMO energy levels of conjugated polymers AB - A series of copolymers of 2,5-dicyano-1,4-phenylenevinylene and 2-methoxy-5-(2' ethylhexyloxy)-1,4-phenylenevinylene were synthesized by Wittig reaction. The HOMO and LUMO energy levels of copolymers can be easily tuned by controlling the feed ratio of co-monomers. PMID- 10840954 TI - Catalysis of acyl group transfer by a double-displacement mechanism: the cleavage of aryl esters catalyzed by calixcrown-Ba2+ complexes AB - The scope of the barium salt of p-tert-butylcalix[4]arene-crown-5 as a transacylation catalyst has been defined by evaluating its efficiency in the methanolysis of a series of aryl acetates at 25.0 degrees C in MeCN/MeOH 9:1 (v/v) under slightly basic conditions. In this system a phenolic hydroxyl is the acyl-receiving and -releasing unit in a double-displacement mechanism. The complexed barium ion acts both as a nucleophile carrier and a built-in Lewis acid in providing electrophilic assistance to the ester carbonyl both in the acylation and deacylation step (nucleophilic-electrophilic catalysis). Turnover capability is ensured by the acylated intermediate reacting with the solvent more rapidly than the original ester, but a serious drawback derives from the incursion of back-acylation of the liberated phenol. A gradual shift from rate-determining deacylation (p-nitrophenyl acetate) to rate-determining acylation (phenyl acetate) is observed along the investigated series. It is shown that the scope of the catalyst is restricted to acetate esters whose reactivity lies in the range approximately defined by the phenyl acetate-p-nitrophenyl acetate pair, with a maximum efficiency for p-chlorophenyl acetate. Moreover, the catalyst effectively promotes ester interchange between phenols, showing that its activity is not limited to solvolysis reactions. The very high sensitivity of the rate of acylation of the catalyst to leaving group basicity has been interpreted as due to rate-determining decomposition of the tetrahedral intermediate, which is believed to arise from the presumably low basicity of the metal ion stabilized nucleophile. The turnover frequency was in the range of 3.8 x 10(-4) min(-1) for phenyl acetate to 7.4 x 10(-3) min(-1) for p-nitrophenyl acetate ([ArOAc]0=4.0 mM]). A first attempt to enhance the rate of acylation of the catalyst through intramolecular general acid catalysis is also described. PMID- 10840955 TI - STM imaging of a heptanuclear ruthenium(II) dendrimer, mono-add layer on graphite AB - Scanning tunneling microscopy (STM) and molecular mechanics calculations were used to investigate the long-range packing and the structure of an heptanuclear ruthenium (II) dendritic species, as a PF6- salt. STM imaging was carried out on a mono-add layer of the ruthenium dendrimer formed by physisorption from a 1,2,4 trichlorobenzene solution at the liquid-graphite interface. The packing of the molecules on the surface was visualised by the formation of ordered patterns and a distance of 27 +/- 2 A was measured between two adjacent lamellae. The comparison of this dimension with the molecular-modelling data indicates that the lamellae were formed by rows of dendrimer molecules in which the counterions (PF6 ) were strongly associated with the Ru atoms. The images acquired with higher spatial resolution revealed the presence of repeating units within the lamellae. The comparison of the STM images with the modelling results allowed the attribution of the repeating units observed in the imaged pattern to the STM signature of single dendrimer molecules. PMID- 10840956 TI - Partially hydroxylated 2,5-disubstituted bis-tetrahydrofurans from carbohydrates AB - The diverse bioactivities of annonaceous acetogenins have recently attracted increasing interest. Many of these natural products contain one or more 2,5 disubstituted tetrahydrofuran rings as a core unit; these are important for the bioactivity, since it is believed that these anchor the compounds to the surface of the membrane. Therefore, the synthesis of functionalized bis-tetrahydrofurans is an important task and we have developed a synthetic pathway to all four diastereomeric, partially hydroxylated bis-tetrahydrofurans, that is, 3,6:7,10) dianhydro-2,8,9-trideoxy-L-erythro-D-ido-undecitol (1), 3,6:7,10-dianhydro-2,8,9 trideoxy-D-threo-D-ido-undecitol (2), 3,6:7,10-dianhydro-2,8,9-trideoxy-L-threo-D ido-undecitol (3), and 3,6:7,10-dianhydro-2,8,9-trideoxy-D-erythro-D-ido undecitol (4) starting from D-glucose. The reaction of the aldose with Meldrum's acid led to the C-glycosidic 3,6-anhydro-1,4-lactone 6, which was converted to the aldehyde building block 2,5-anhydro-3,4,7-tri-O-benzyl-6-deoxy-aldehydo-D-ido heptose (11). Chain elongation of 11 with the Grignard reagent derived from 1 bromo-3-butene gave the diastereomers 3,6-anhydro-1,4,5-tri-O-benzyl-2,8,9,10,11 pentadeoxy-L-glycero-D-ido-undec-10-enitol (12) and 3,6-anhydro-1,4,5-tri-O benzyl-2,8,9,10,11-pentadeoxy-D-glycero-D-ido-undec-10-enitol (13). The relative threo configuration of the major product 12 was confirmed by X-ray structure analysis. Epoxidation and subsequent cyclization afforded the cis and trans diastereomers 19 and 20, respectively, in a 1:1 ratio. Subsequent cleavage of the protecting groups and separation of the isomers furnished the target compounds in good overall yields. PMID- 10840957 TI - Quantitative chirality/enantioselectivity relations in large random supramolecular structures AB - We study the relationship between shape and enantioselectivity, employing quantitative geometric chirality measurements. The model we use comprises of the boundary surfaces of two-dimensional (2D) chiral, large, random selectors (diffusion limited aggregates), interacting with homologous series of small 2D chiral S-shaped probes (the selectands). We show how the enantioselectivity of the selectors depends on the chirality of the selectands and report the following findings: I) The enantioselectivity of a chiral selector can switch preference from the "right" to the "left" enantiomer within a homologous series of selectands. II) At this switch point the chiral selector is functionally achiral. III) Within a homologous series of chiral selectands, there is a "resonance of recognition", namely, the classical key-lock concept is replaced by a picture of various degrees of recognition. IV) The degree of enantioselectivity and the switch in handedness preference are the outcome of a complex interplay between the details of the specific geometry of the selector and the selectand, and the global shape parameter of chirality measure. V) It is shown that isochiral selectands, namely selectands of the same chirality value, may be recognized differently by a chiral selector. VI) It is proposed that a more realistic way to treat the issue of minimal points needed for chiral interaction is resolution based. VII) It is shown how to attach handedness to purely random objects. PMID- 10840959 TI - Excision of the AB - The synthesis of new molybdenum cluster selenocyanide anionic complexes [Mo6Se8(CN)6]7- and [Mo6Se8(CN)6]6- is reported. The [Mo6Se8(CN)6]7- ion was obtained by excision of the cluster core [Mo6Se8] from a Chevrel phase in the reaction of Mo6Se8 with KCN at 650 degrees C; the [Mo6Se8(CN)6]6- ion is formed by oxidation of [Mo6Se8(CN)6]7-. New cluster salts K7[Mo6Se8(CN)6] x 8H2O (1) and (Me4N)4K2[Mo6Se8(CN)6] x 10H2O (2) were isolated and their crystal structures were solved. Compound 1 crystallizes in the cubic space group Fm3m (a=15.552(2) A, Z=4, V=3761.5(8) A3), compound 2 crystallizes in the triclinic space group P1 (a=11.706(2), b=11.749(2), c=12.459(2) A, alpha=72.25(1), beta=77.51(1), gamma=63.04(1), Z=1, V=1448.5(4) A3). Compound 1 is paramagnetic due to an availability of 21 electrons per Mo6 cluster; cyclic voltammetry reveals a quasi reversible transition [Mo6Se8(CN)6]7- <--> [Mo6Se8(CN)6]6-, E1/2=0.63 V. PMID- 10840958 TI - Oxepinamides A-C and fumiquinazolines H--I: bioactive metabolites from a marine isolate of a fungus of the genus Acremonium. AB - Three new oxepin-containing natural products (1-3) and two new fumiquinazoline metabolites (4-5) have been isolated from organic extracts of the culture broth and mycelia of an Acremonium sp., a fungus obtained from the surface of the Caribbean tunicate Ecteinascidia turbinata. The structures of the five compounds were determined through extensive analysis of 1D- and 2D-NMR data, and mass spectrometry. Compound 1 exhibited good anti-inflammatory activity in a topical RTX-induced mouse ear edema assay. Compounds 4 and 5 exhibited weak antifungal activity toward Candida albicans in a broth microdilution assay. PMID- 10840960 TI - A second generation synthesis of the MBr1 (globo-H) breast tumor antigen: new application of the n-pentenyl glycoside method for achieving complex carbohydrate protein linkages. AB - A new synthesis of the hexasaccharide MBr1 antigen (globo-H) is reported. A revised construction with improved efficiency was necessary because an anti cancer vaccine containing this antigen is entering phase II and phase III clinical trials for prostate cancer. The key feature of this second generation synthesis is the preparation of globo-H as its n-pentenyl glycoside. This group serves as an anomeric protecting group and as a linker for bioconjugation to carrier protein. The resultant synthesis allows for the production of suitable quantities of globo-H for clinical trials. PMID- 10840961 TI - Facilitated transport of salts by neutral anion carriers AB - Partitioning of ions from water to the membrane solvent (NPOE) can be quantified by Gibbs free energies of transfer, deltaG(tr,NPOE)(ion). These were derived from transport studies of lipophilic salts through supported liquid membranes (SLMs) in the absence of the carrier. Partition coefficients Kp for various salts can now be calculated. The neutral anion receptors uranyl sal(oph)enes 1-5 transport Cl- and H2PO4- as tetrapropylammonium salts. The transport is diffusion-limited and can be described by two transport parameters Dm and K(ex). From the extraction constants K(ex) and the partition coefficients Kp of the transported salts, the association constants Ka of the anion receptors for Cl- and H2PO4- in NPOE were determined. Competitive transport with carriers 3 and 4 of NPr4H2PO4 and NPr4Cl demonstrated highly selective transport of H2PO4- even in the presence of excess of Cl-. PMID- 10840962 TI - Ultrafast dynamics of photochemical radical formation from AB - The excited-state dynamics and photochemistry of [Re(R)(CO)3(dmb)] (R=Me, Et); dmb=4,4'-dimethyl-2,2'-bipyridine) in CH2Cl2 have been studied by time-resolved visible absorption spectroscopy on a broad time scale ranging from approximately 400 fs to a few microseconds, with emphasis on the femtosecond and picosecond dynamics. It was found that the optically prepared Franck-Condon 1MLCT (singlet metal-to-ligand charge transfer) excited state of [Re(R)(CO)3(dmb)] undergoes femtosecond branching between two pathways (< or =400 fs for R=Me; approximately 800 fs for R=Et). For both methyl and ethyl complexes, evolution along one pathway leads to homolysis of the Re-R bond via a 3SBLCT (triplet sigma-bond-to ligand charge transfer) excited state, from which [Re(S)(CO)3(dmb)]* and R* radicals are formed. The other pathway leads to an inherently unreactive 3MLCT state. For [Re(Me)(CO)3(dmb)], the 3MLCT state lies lowest in energy and decays exclusively to the ground state with a lifetime of approximately 35 ns, thereby acting as an excitation energy trap. The reactive 3SBLCT state is higher in energy. The quantum yield (0.4 at 293 K) of the radical formation is determined by the branching ratio between the two pathways. [Re(Et)(CO)3(dmb)] behaves differently: branching of the Franck-Condon state between two pathways still occurs, but the 3MLCT excited state lies above the dissociative 3SBLCT state and can decay into it. This shortens the 3MLCT lifetime to 213 ps in CH2Cl2 or 83 ps in CH3CN. Once populated, the 3SBLCT state evolves toward radical photoproducts [Re(S)(CO)3(dmb)]* and Et*. Thus, population of the 3MLCT excited state of [Re(Et)(CO)3(dmb)] provides a second, delayed pathway to homolysis. Hence, the quantum yield is unity. The photochemistry and excited-state dynamics of [Re(R)(CO)3(dmb)] (R=Me, Et) complexes are explained in terms of the relative ordering of the Franck-Condon, 3MLCT, and 3SBLCT states in the region of vertical excitation and along the Re-R reaction coordinate. A qualitative potential energy diagram is proposed. PMID- 10840963 TI - Spin-spin coupling tensors in fluoromethanes AB - All spin-spin coupling tensors J of the fluoromethanes CH3F, CH2F2, and CHF3 are obtained theoretically by multiconfiguration self-consistent field linear response (MCSCF LR) ab initio calculations. Furthermore the principal values and the orientation of the principal axis systems of each theoretical J tensor are specified. Experimental liquid crystal NMR (LC NMR) data on the tensorial properties of the CF spin spin coupling in CH3F and CH2F2, and the FF spin-spin coupling in CHF3 are also reported. In the analysis of the experiments, the contributions from molecular vibrations, as well as that of the correlation of vibrational and rotational motion to the experimental anisotropic couplings, D(exp), are taken into account. The information of the anisotropic indirect coupling, 1/2J(aniso), is detected as the difference between D(exp) and the calculated dipolar coupling, D(calc). The extracted indirect contributions, 1/2J(aniso), are in fair agreement with the ab initio results. All relative (experimental and theoretical) CF and FF indirect contributions, 1/2J(aniso)/D(exp), are negative and under 1.7% in magnitude, when the observed molecular orientations are used. Therefore, in the one bond CF couplings and in the two bond FF couplings, the indirect contribution can normally be ignored without introducing serious error to the determination of molecular orientation and/or structure. However, a more accurate method is to partially correct for the indirect contribution by utilising the transferability of the spin-spin coupling tensors in related molecules. This is due to the fact that even small contributions may be significant, if the order parameter of the internuclear direction is negligibly small, leading to dominating indirect contributions. The very good agreement of the experimental values with the calculated coupling constants and the reasonable agreement in the anisotropic properties, which are experimentally much more difficult to define, indicates that the MCSCF LR method is capable of producing reliable J tensors for these systems, contrary to the case of density-functional theory. PMID- 10840964 TI - Elucidating the mode of action of a corrosion inhibitor for iron AB - Two polymetallic iron(III) complexes 1 and 2 have been synthesised from the known corrosion inhibitor 3-(4-methylbenzoyl)-propionic acid HL1 and their crystal structures determined. Coordination geometries extracted from these structures have been used as the basis for molecular modelling onto idealised iron(III) oxide surfaces as an aid to understanding the efficacy of inhibitors of the 4 keto acid type. The proposed mode of action involves 1,3-bridging didentate coordination of the carboxylate function of L1 to two FeIII ions, hydrogen-bond formation between the 4-keto group of L1 and a bridging surface hydroxy group, as well as close packing of the aromatic end groups, which should generate a hydrophobic barrier on the surface. Adsorption isotherm experiments have been used to compare the strengths of binding of related carboxylic acids onto iron(III) oxide surfaces and indicate that the presence of the 4-keto function leads to the formation of significantly more stable surface complexes. PMID- 10840965 TI - Synthesis of high-mannose type neoglycolipids: active targeting of liposomes to macrophages in gene therapy. AB - The concise synthesis of five biantennary oligomannose neoglycolipids is presented. Employing a strategy based on the principles of reactivity tuning and orthogonal activation, the oligomannose moieties, isolated from the glycoprotein 63 of the parasite Leishmania mexicana amazonensis, were rapidly assembled taking advantage of common structural motifs found in these N-glycans. Deprotection of all structures was achieved in high yield by hydrogenolysis. The deprotected glycoconjugates were subsequently coupled to a cholesteroldiamine derivative using diethylsquarate as a linker. The resulting neoglycolipids will be used as additives to cationic liposome formulations in the active targeting of liposomes to macrophages. PMID- 10840966 TI - Diagnostic organometallic and metallodendritic materials for SO2 gas detection: reversible binding of sulfur dioxide to arylplatinum(II) complexes AB - A series of square-planar platinum(II) complexes of the N,C,N'-terdentate coordinating monoanionic "pincer" ligand, [PtX(4-E-2,6-[CH2NRR']2-C6H2](X=Cl, Br, I, tolyl; R, R'=Et, Me; E=H, OH, OSiMe2tBu) has been prepared. In the presence of sulfur dioxide, these complexes spontaneously adsorb this gas to form penta coordinated adducts. Solid-state crystal-structure analyses of the SO2 adducts 8c (X=I, R=R=Me, E=OSiMe2tBu) and 11 (X=Cl, R=R'=Me, E=OH) show a square-pyramidal geometry around the metal center with SO2 in the apical position. Most interestingly. the adduct 11 forms similar Pt-Cl... H-O hydrogen-bonded alpha type networks as the corresponding SO2-free complex 5. The conservation of the supramolecular information (hydrogen-bonded self-assembly) throughout a reaction (SO2 adsorption) is unprecedented in crystal engineering. Adduct formation in the solid state or in solution is fast and reversible and is indicated by a characteristic color change of the material from colorless to bright orange. Since facile methods have been developed to remove SO2 from the adducts and to regenerate the square-planar starting complexes, these complexes fulfill several essential prerequisites of sensor materials for repeated diagnostic SO2 detection. The platinum sensors have been found to be highly selective for sulfur dioxide and particularly sensitive for submilimolar to molar gas quantities. Their response capacity is tuneable by electronic and steric modifications of the ligand array by introduction of, for example, different substituents on the nitrogen donors. The periphery of dendrimers is shown to be an appropriate macromolecular support for anchoring the detection-active sites, thus allowing full recovery of the sensor materials for repeated use. By using this concept, metallo-dendrimers 3 and 15 have been prepared. Owing to the dendritic connectivity, these sensors are suitable for repetitive qualitative and quantitative detection of small amounts of SO2. PMID- 10840967 TI - A valence bond study of the Bergman cyclization: geometric features, resonance energy, and nucleus-independent chemical shift (NICS) values AB - The Bergman cyclization of (Z)-hex-3-ene-1,5-diynes (1, enediynes), which produces pharmacologically important DNA-cleaving biradicals (1,4-benzyne, 2), was studied by using Hartree-Fock (HF) and density-functional theory (DFT) based valence bond (VB) methods (VB-HF and VB-DFT, respectively). We found that only three VB configurations are needed to arrive at results not too far from complete active space [CASSCF(6 x 6)] computations, while the quality of VB-DTF utilizing the same three configurations improves upon CASSCF(6 x 6) analogous to CASPT2. The dominant VB configuration in 1 contributes little to 2, while the most important biradical configuration in 2 plays a negligible role in 1. The avoided crossing of the energy curves of these two configurations along the reaction coordinate leads to the transition state (TS). As a consequence of the shape and position of the crossing section, the changes in geometry and in the electronic wavefunction along the reaction coordinate are non-synchronous; the TS is geometrically approximately 80% product-like and electronically approximately 70% reactant-like. While the pi resonance in the TS is very small, it is large (64.4 kcal mol(-1)) for 2 (cf. benzene=61.5 kcal mol(-1)). As a consequence, substituents operating on the sigma electrons should be much more effective in changing the Bergman reaction cyclization barrier. Furthermore, additional sigma resonance in 2 results in unusually high values for the nucleus-independent chemical shift (NICS, a direct measure for aromaticity). Similarly, the high NICS value of the TS is due mostly to sigma resonance to which the NICS procedure is relatively sensitive. PMID- 10840968 TI - Biomimetic synthesis of lantibiotics. AB - The lantibiotics are a class of highly posttranslationally modified small peptide antibiotics containing numerous lanthionine and dehydroamino acid residues. We have prepared peptides containing multiple dehydroamino acids and cysteine residues in order to probe the biomimetic synthesis of the lantibiotics from their precursor peptides. A novel synthetic methodology was developed to allow the synthesis of multiple dehydroamino acid containing peptides. Cyclisations were rapid, quantitative and regiospecific. Remarkably the peptide sequences alone appear to contain sufficient information to direct a series of stereo- and regiospecific ring closures. Thus both the two linear peptides for the B and E rings closed stereoselectively. In the case of the A-ring precursor peptide which contained two dehydroamino acids, cyclisation was again totally regioselective, although not totally stereoselective. PMID- 10840969 TI - Ring opening of 1-methylcyclopropene and cyclopropene: matrix infrared spectroscopic identification of 2-butene-1,3-diyl and propene-1,3-diyl AB - Triplet 2-butene-1,3-diyl (T-11) was generated on irradiation of 1 methylcyclopropene (10) in a bromine-doped xenon matrix and was characterized by means of IR spectroscopy for the first time. Experimental results suggest that triplet propene-1,3-diyl (T-3) is formed from cyclopropene (1) under similar conditions. In accordance with theoretical calculations, the experimental data indicate that the reactions 1-->3 and 10-->11 are the lowest energy ground-state pathways for the ring opening of 1 and 10, respectively. PMID- 10840970 TI - Oxidative addition of palladium(0) complexes generated from AB - The major complex formed in solution from [[Pd0(dba)2]+1P-N] mixtures is [Pd0(dba)(P-N)] (dba=trans,trans-dibenzylideneacetone; P-N=PhPN, 1-dimethylamino 2-diphenylphosphinobenzene; FcPN, N,N-dimethyl-1-[2 (diphenylphosphino)ferrocenyl]methylamine; OxaPN, 4,4'-dimethyl-2-(2 diphenylphosphinophenyl)-1,3-oxazoline). Each complex consists of a mixture of isomers involved in equilibria: two 16-electron rotamer complexes [Pd0(eta2 dba)(eta2-P-N)] and one 14-electron complex [Pd0(eta2-dba)(eta1-P-N)] observed for FcPN and OxaPN. [Pd0(dba)(PhPN)] and [SPd0(PhPN)] (S solvent) react with PhI in an oxidative addition: [SPd0(PhPN)] is intrinsically more reactive than [Pd0(dba)(PhPN)]. This behavior is similar to that of the bidentate bis-phosphane ligands. When the PhPN ligand is present in excess, it behaves as a monodentate phosphane ligand, since [Pd0(eta2-dba)(eta1-PhPN)2] is formed first by preferential cleavage of the Pd-N bond instead of the Pd olefin bond. [Pd0(eta1 PhPN)3] is also eventually formed. [Pd0(dba)(FcPN)] and [Pd0(dba)(OxaPN)] are formed whatever the excess of ligand used. [SPd0(FcPN)] and [SPd0)(OxaPN)] are not involved in the oxidative addition. The 16-electron complexes [Pd0(eta2 dba)(eta2-FcPN)] and [Pd0(eta2-dba)(eta2-OxaPN)] are found to react with PhI via a 14-electron complex as has been established for [Pd0(eta2-dba)(eta1-OxaPN)]. Once again, the cleavage of the Pd-N bond is favored over that of Pd-olefin bond. This work demonstrates the higher affinity for [Pd0(P-N)] of dba compared with the P-N ligand, and emphasizes once more the important role of dba, which either controls the concentration of the most reactive complex, [SPd0(PhPN)], or is present in the reactive complexes, [Pd0(dba)(FcPN)] or [Pd0(dba)(OxaPN)], and thus contributes to their intrinsic reactivity. PMID- 10840971 TI - Benzoylformate decarboxylase from Pseudomonas putida as stable catalyst for the synthesis of chiral 2-hydroxy ketones. AB - The thiamin diphosphate- and Mg2+-dependent enzyme benzoylformate decarboxylase (BFD) from Pseudomonas putida was characterized with respect to its suitability to catalyze the formation of chiral 2-hydroxy ketones in a benzoin-condensation type reaction. Carboligation constitutes a side reaction of BFD, whereas the predominant physiological task of the enzyme is the non-oxidative decarboxylation of benzoylformate. For this purpose the enzyme was obtained in sufficient purity from Pseudomonas putida cells in a one-step purification using anion-exchange chromatography. To facilitate the access to pure BFD for kinetical studies, stability investigations, and synthetical applications, the coding gene was cloned into a vector allowing the expression of a hexahistidine fusion protein. The recombinant enzyme shows distinct activity maxima for the decarboxylation and the carboligation beside a pronounced stability in a broad pH and temperature range. The enzyme accepts a wide range of donor aldehyde substrates which are ligated to acetaldehyde as an acceptor in mostly high optical purities. The enantioselectivity of the carboligation was found to be a function of the reaction temperature, the substitution pattern of the donor aldehyde and, most significantly, of the concentration of the donor aldehyde substrate. Our data are consistent with a mechanistical model based on the X-ray crystallographic data of BFD. Furthermore we present a simple way to increase the enantiomeric excess of (S)-2-hydroxy-1-phenyl-propanone from 90% to 95% by skillful choice of the reaction parameters. Enzymatic synthesis with BFD are performed best in a continuously operated enzyme membrane reactor. Thus, we have established a new enzyme tool comprising a vast applicability for stereoselective synthesis. PMID- 10840972 TI - Application of P-stereogenic aminophosphine phosphinite ligands in asymmetric hydroformylation AB - New chiral aminophosphine phosphinite ligands with a stereogenic center at the aminophosphine phosphorus atom were prepared based on (R,S)-ephedrine as the chiral auxiliary and backbone. Substituents at the chiral aminophosphine as well as at the phosphinite phosphorus atom were varied. These new ligands were applied to the rhodium-catalyzed asymmetric hydroformylation of vinyl arenes. The enantiomeric excess reached up to 77%. 1H and 31P NMR studies of the Rh complexes under syngas pressure reveal that [HRh(CO)2(PP)] complexes with the NP* moiety in an axial position are responsible for enantioselectivity. PMID- 10840973 TI - The butterfly Danaus chrysippus is infected by a male-killing Spiroplasma bacterium. AB - Many insects carry maternally inherited bacteria which kill male offspring. Such bacteria will spread if male death benefits the female siblings who transmit the bacterium, and they are therefore expected in insects with antagonistic sibling interactions. We report that the butterfly Danaus chrysippus is host to a maternally inherited male-killing bacterium. Using diagnostic PCR and rDNA sequence, the bacterium was identified as a Spiroplasma closely related to 2 ladybird beetle male-killers and the tick symbiont Spiroplasma ixodetis. The male killer was found to have a geographically restricted distribution, with up to 40% of females being infected in East Africa, but no detectable infection in small samples from other populations. Danaus chrysippus is a surprising host for a male killer as its eggs are laid singly. This suggests that the ecological conditions permitting male-killers to invade may be more widespread than previously realized. PMID- 10840974 TI - Chronic Plasmodium falciparum infections in an area of low intensity malaria transmission in the Sudan. AB - Chronic Plasmodium falciparum malaria infections in a Sudanese village, in an area of seasonal and unstable malaria transmission, were monitored and genetically characterized to study the influence of persistent infection on the immunology and epidemiology of low endemicity malaria. During the October December malaria season of 1996, 51 individuals out of a population of 420 had confirmed and treated P. falciparum malaria in the village of Daraweesh in eastern Sudan. In a cross-sectional survey carried out in December 1996, an additional 6 individuals were found to harbour a microscopically negative but polymerase chain reaction (PCR)-positive P. falciparum infection. On 1 January 1997, a cohort of 43 individuals aged from 9 to 53, recruited from this group of recently malaria-infected individuals agreed to donate fortnightly blood samples for the next 9 months, the first 6 of which constitute the long Sudanese dry season when transmission falls to undetectable levels. Each blood sample was tested for the presence of persistent malaria infection by microscopy and PCR. Parasite-positive samples were genotyped using PCR assays that detect allelic polymorphism at the MSP-1, MSP-2 and GLURP marker gene loci. Of 43 individuals 16 were found to maintain chronic P. falciparum infections which were continuously genetically characterized. PMID- 10840975 TI - Molecular and phylogenetic analysis of Cryptosporidium muris from various hosts. AB - Isolates of Cryptosporidium muris and C. serpentis were characterized from different hosts using nucleotide sequence analysis of the rDNA 18S and ITS1 regions, and the heat-shock (HSP-70) gene. Phylogenetic analysis confirmed preliminary evidence that C. muris is not a uniform species. Two distinct genotypes were identified within C. muris; (1) C. muris genotype A; comprising bovine and camel isolates of C. muris from different geographical locations, and (2) C. muris genotype B comprising C. muris isolates from mice, a hamster, a rock hyrax and a camel from the same enclosure. These 2 genotypes may represent separate species but further biological and molecular studies are required for confirmation. PMID- 10840976 TI - Cellular and humoral immune response of European sea bass (Dicentrarchus labrax L.) (Teleostei: Serranidae) immunized with Sphaerospora dicentrarchi (Myxosporea: Bivalvulida). AB - The immune response of European sea bass after intracaelomic immunization with Sphaerospora dicentrarchi was studied. Fish were injected with S. dicentrarchi spores (DIC), with spores plus adjuvant (DIC + FCA), with adjuvant alone (FCA) or with PBS. Several parameters of the immune response were measured. Serum lysozyme increased significantly in DIC fish 1 week after immunization (p.i.) and it remained significantly higher in DIC + FCA fish 4 weeks p.i., and in DIC fish 8 weeks p.i. than in PBS-injected fish. The number of nitroblue tetrazolium positive blood cells was significantly higher in DIC + FCA fish 1, 4 and 8 weeks p.i, but the highest values were detected 1 week p.i. The highest stimulation index was detected in phagocytes from DIC + FCA fish. The number of S. dicentrarchi antibody-secreting cells was significantly higher in DIC + FCA fish than in DIC fish. Serum from DIC and DIC + FCA fish, stained the polar capsules and the valves of S. dicentrarchi spores in immunohistochemistry. Serum antibodies could not be detected using immunoblot assay. All these results show that immunization with S. dicentrarchi resulted in the activation of the non specific immune response, mainly 7 days p.i. A specific humoral response against the parasite was also demonstrated but it had a low magnitude. PMID- 10840977 TI - Host-finding in Echinostoma caproni: miracidia and cercariae use different signals to identify the same snail species. AB - The snail host signals releasing host-finding responses in miracidia and cercariae of Echinostoma caproni were analysed by fractionation of snail conditioned water (SCW). Cercariae responded non-specifically to organic and hydrophilic, low molecular weight components of SCW showing their typical turning response. Hydrolysis of peptides in SCW had no effect on cercarial responses. An artificial mixture of amino acids in concentrations determined from SCW as well as glycine alone in a concentration corresponding to the total concentration of amino acids in SCW showed nearly the same efficacy as SCW itself. Miracidia responded to a high molecular weight glycoprotein fraction, which could be isolated from SCW by ion-exchange and size-exclusion chromatography. In contrast to an Egyptian Schistosoma mansoni strain, the echinostome miracidia were not able to differentiate between different snail species. The results show for the first time that miracidia and cercariae of the same species may use different signals to identify the same snail host species. This indicates an independent evolution of host-finding mechanisms in the two parasite stages. PMID- 10840978 TI - Phylogenetic relationships of human and wildlife piroplasm isolates in the western United States inferred from the 18S nuclear small subunit RNA gene. AB - The 18S nuclear small subunit ribosomal RNA gene of piroplasms from wildlife and human cases of babesiosis in the western USA were isolated by PCR and sequenced. Phylogenetic analyses of these sequences and comparisons with sequences from other Babesia and Theileria species revealed that piroplasm isolates from the human cases were indistinguishable from some of the isolates from the western wildlife species, most notably the isolates from mule deer (Odocoileus hemionus). These results suggest that large ungulates may serve as reservoirs for human piroplasm infection. The western piroplasm isolates from humans and wildlife formed a distinct clade, separate from other piroplasms found worldwide. PMID- 10840979 TI - Changes of mate occur in Schistosoma mansoni. AB - Male and female schistosomes are generally assumed to form stable monogamous pairs for the whole span of their long existence in the mammalian host. Recent evidence from mixed infections has shown that Schistosoma mansoni males can displace S. intercalatum males from their homologous partners, but no information exists about the existence of similar phenomena within a single schistosome species. Here, we determine whether male S. mansoni can displace males of the same species from pre-formed pairs in vivo. The availability of clear-cut genetic markers of drug resistance in schistosomes was exploited to show that hycanthone sensitive S. mansoni males can displace homospecific hycanthone resistant males from pre-formed pairs and vice versa. The frequency of changes is dependent on the magnitude of the excess single males competing with paired worms. The possible mechanics and the biological significance of mate changing are discussed. PMID- 10840981 TI - Population structure and genetic typing of Trypanosoma cruzi, the agent of Chagas disease: a multilocus enzyme electrophoresis approach. AB - A set of 434 Trypanosoma cruzi stocks from a wide ecogeographical range was analysed by Multilocus Enzyme Electrophoresis for 22 genetic loci. Strong linkage disequilibrium, not associated with geographical distance, and 2 main genetic clusters each considerably heterogeneous, was observed. These results support the hypotheses previously proposed that T. cruzi natural populations are composed of highly diversified genetic clones distributed into 2 main phylogenetic lineages: lineage 1, the most ubiquitous in the endemic area, was more frequently observed in sylvatic cycles, whereas lineage 2, predominant in humans and domestic cycles, in the southern part of the area surveyed, was further partitioned into 5 lesser genetic subdivisions. T. cruzi appears therefore subdivided into at least 6 'discrete typing units' or DTUs (Tibayrenc, 1998a-c). We have identified various specific isoenzyme markers ('tags'; Tibayrenc, op. cit.) suitable for the routine identification of these DTUs for epidemiological tracking purposes. We discuss the correspondence with previous classifications and with the recent recommendations of the 90th anniversary of the discovery of Chagas disease symposium, as well as the impact of T. cruzi genetic variability on this parasite's biomedical diversity. PMID- 10840980 TI - Schistosoma mansoni phosphofructokinase: immunolocalization in the tegument and immunogenicity. AB - Schistosoma mansoni depends for its survival on glycolysis. Two glycolytic enzymes, glyceraldehyde-3P-dehydrogenase and triose-phosphate dehydrogenase, found in both the adult and schistosomular tegument, have been reported to confer partial protection against cercarial infection. This paper describes the immunogenic properties of phosphofructokinase (PFK), a rate-limiting enzyme of glycolysis, and its localization in the tegument and adjacent tissues. Recombinant schistosome PFK was used as antigen. A polyclonal antibody against purified PFK from Fasciola hepatica was affinity purified using recombinant PFK and used in combination with immunogold labelling to identify PFK by transmission electron microscopy in cryosections. In both adult worms and in schistosomula most immunogold label localized in the cytoplasmic syncytial region with less being found in the tegument. There was no significant PFK localization within or external to the outer membrane. Sera from mice immunized with recombinant S. mansoni PFK with Freund's adjuvant or alum plus rIL-12 demonstrated high titres of anti-PFK IgG, but no protection against cercarial infection. Sera from mice that were acutely or chronically infected or multiply exposed to irradiated cercariae did not recognize recombinant schistosome PFK in either Western blotting or ELISA. Similarly, sera from humans infected with S. mansoni did not recognize PFK. We conclude that in spite of the high immunogenicity of rPFK in mice, it is not a significant immunogen during the course of infection and does not confer protection from schistosomiasis. One main difference between PFK and the other 2 glycolytic enzymes seems to be the inaccessibility of PFK to the outside surface of the tegument. PMID- 10840982 TI - Single-stranded endonuclease activity in the excretory--secretory products of Trichinella spiralis and Trichinella pseudospiralis. AB - A novel acidic extracellular single-stranded endonuclease was demonstrated for the first time in the excretory-secretory (E-S) products of 2 species of Trichinella. Unlike the double-stranded endonuclease reported earlier, the single stranded molecule is divalent cation independent and is detected in both T. spiralis and T. pseudospiralis E-S products. It hydrolysed single-stranded DNA and RNA at comparable rates. The single-stranded endonuclease was sensitive to inhibition by Zn2+ and to high concentrations of NaCl. Zymographic analysis indicated that it was encoded by at least 3 peptides of Mr approximately 50-60 kDa. The rate of hydrolysis of single-stranded targets by the E-S products was substantially higher than that of the double-stranded molecule. Due to the differences in peptide profile, divalent cation dependence, and species-specific expression, the single and double-stranded endonucleases are likely to be encoded by different proteins and may have different functions. PMID- 10840983 TI - L-glutamate binding sites of parasitic nematodes: an association with ivermectin resistance? AB - Nematode membrane preparations contain high amounts of low-affinity specific L glutamate binding sites. The numbers of these sites were increased in 2 isolates, one field-derived and the other laboratory-derived, of ivermectin-resistant Haemonchus contortus and a field isolate of ivermectin-resistant Telodorsagia circumcincta, when compared to control, drug-sensitive isolates. Specific [3H]ivermectin binding to these membrane preparations showed no differences between ivermectin-sensitive and resistant isolates and the number of ivermectin binding sites was approximately 100-fold less than the number of L-glutamate binding sites. Kinetic analysis of L-glutamate binding suggested the presence of at least 2 classes of binding site. L-Glutamate binding was blocked by ibotenic acid, kynurenic acid and beta-hydroxyaspartate, but not by ivermectin, argiopine, kainate, quisqualate or NMDA. Competition assays with ibotenic acid suggested that there were 2 distinct populations of glutamate binding sites and that the site with the lower affinity for ibotenate was upregulated in the ivermectin resistant nematodes. In the field isolate of resistant H. contortus we found no coding changes in the cDNAs encoding glutamate-gated chloride channel subunits HG2, HG3 and HG4, nor were any changes in channel expression detected using subunit-specific antibodies. The low-affinity binding site is unlikely to be associated with the ivermectin receptor in these nematodes. PMID- 10840984 TI - Asymptomatic bacteriuria in diabetic women. PMID- 10840985 TI - Does insulin play a role in cardiovascular autonomic regulation? PMID- 10840986 TI - Vitamin C and hyperglycemia in the European Prospective Investigation into Cancer -Norfolk (EPIC-Norfolk) study: a population-based study. AB - OBJECTIVE: To examine the cross-sectional association between plasma vitamin C, self-reported diabetes, and HbA1c. RESEARCH DESIGN AND METHODS: Data from a population-based study of diet, cancer, and chronic disease were analyzed. A total of 2,898 men and 3,560 women 45-74 years of age who were registered with general practices in Norfolk, U.K., were recruited to the European Prospective Investigation Into Cancer-Norfolk study between 1995 and 1998. RESULTS: Mean plasma vitamin C levels were significantly higher in individuals with HbA1c levels < 7% than in those with self-reported diabetes or prevalent undiagnosed hyperglycemia (HbA1c > or = 7%). An inverse gradient of mean plasma vitamin C was found in both sexes across quintiles of HbA1c distribution < 7%. The odds ratio (95% CI) of having prevalent undiagnosed hyperglycemia per 20 micromol/l (or 1 SD) increase in plasma vitamin C was 0.70 (0.52-0.95) (adjusted for sex, age, BMI, waist-to-hip ratio, tertiary education, any use of dietary supplements, vegetarian diet, alcohol consumption, physical activity, dietary vitamin E, dietary fiber, dietary saturated fat, and smoking history). The unadjusted change in HbA1c per 20 micromol/l increase in vitamin C estimated by linear regression was -0.12% (-0.14 to -0.09) in men and -0.09% (-0.11 to -0.07) in women. After adjusting for the possible confounders, these values were -0.08% (-0.11 to -0.04) in men and -0.05% (-0.07 to -0.03) in women. CONCLUSIONS: An inverse association was found between plasma vitamin C and HbA1c. Dietary measures to increase plasma vitamin C may be an important public health strategy for reducing the prevalence of diabetes. PMID- 10840987 TI - Effect of supplementation with tomato juice, vitamin E, and vitamin C on LDL oxidation and products of inflammatory activity in type 2 diabetes. AB - OBJECTIVE: To compare the effects of short-term dietary supplementation with tomato juice, vitamin E, and vitamin C on susceptibility of LDL to oxidation and circulating levels of C-reactive protein (C-RP) and cell adhesion molecules in patients with type 2 diabetes. RESEARCH DESIGN AND METHODS: There were 57 patients with well-controlled type 2 diabetes aged <75 years treated with placebo for 4 weeks and then randomized to receive tomato juice (500 ml/day), vitamin E (800 U/day), vitamin C (500 mg/day), or continued placebo treatment for 4 weeks. Susceptibility of LDL to oxidation (lag time) and plasma concentrations of lycopene, vitamin E, vitamin C, C-RP, vascular cell adhesion molecule 1, and intercellular adhesion molecule 1 were measured at the beginning of the study, after the placebo phase, and at the end of the study. RESULTS: Plasma lycopene levels increased nearly 3-fold (P = 0.001), and the lag time in isolated LDL oxidation by copper ions increased by 42% (P = 0.001) in patients during supplementation with tomato juice. The magnitude of this increase in lag time was comparable with the corresponding increase during supplementation with vitamin E (54%). Plasma C-RP levels decreased significantly (-49%, P = 0.004) in patients who received vitamin E. Circulating levels of cell adhesion molecules and plasma glucose did not change significantly during the study. CONCLUSIONS: This study indicates that consumption of commercial tomato juice increases plasma lycopene levels and the intrinsic resistance of LDL to oxidation almost as effectively as supplementation with a high dose of vitamin E, which also decreases plasma levels of C-RP, a risk factor for myocardial infarction, in patients with diabetes. These findings may be relevant to strategies aimed at reducing risk of myocardial infarction in patients with diabetes. PMID- 10840988 TI - The Diabetes Empowerment Scale: a measure of psychosocial self-efficacy. AB - OBJECTIVE: The purpose of this study was to assess the validity, reliability, and utility of the Diabetes Empowerment Scale (DES), which is a measure of diabetes related psychosocial self-efficacy. RESEARCH DESIGN AND METHODS: In this study (n = 375), the psychometric properties of the DES were calculated. To establish validity, DES subscales were compared with 2 previously validated subscales of the Diabetes Care Profile (DCP). Factor and item analyses were conducted to develop subscales that were coherent, meaningful, and had an acceptable coefficient alpha. RESULTS: The psychometric analyses resulted in a 28-item DES (alpha = 0.96) with 3 subscales: Managing the Psychosocial Aspects of Diabetes (alpha = 0.93), Assessing Dissatisfaction and Readiness To Change (alpha = 0.81), and Setting and Achieving Diabetes Goals (alpha = 0.91). Consistent correlations in the expected direction between DES subscales and DCP subscales provided evidence of concurrent validity. CONCLUSIONS: This study provides preliminary evidence that the DES is a valid and reliable measure of diabetes-related psychosocial self-efficacy. The DES should be a useful outcome measure for various educational and psychosocial interventions related to diabetes. PMID- 10840989 TI - Asymptomatic bacteriuria may be considered a complication in women with diabetes. Diabetes Mellitus Women Asymptomatic Bacteriuria Utrecht Study Group. AB - OBJECTIVE: To study the prevalence of and risk factors for asymptomatic bacteriuria (ASB) in women with and without diabetes. RESEARCH DESIGN AND METHODS: A total of 636 nonpregnant women with diabetes (type 1 and type 2) who were 18-75 years of age and had no abnormalities of the urinary tract, and 153 women without diabetes who were visiting the eye and trauma outpatient clinic (control subjects) were included. We defined ASB as the presence of at least 10(5) colony-forming units/ml of 1 or 2 bacterial species in a culture of clean voided midstream urine from an individual without symptoms of a urinary tract infection (UTI). RESULTS: The prevalence of ASB was 26% in the diabetic women and 6% in the control subjects (P < 0.001). The prevalence of ASB in women with type 1 diabetes was 21%. Risk factors for ASB in type 1 diabetic women included a longer duration of diabetes, peripheral neuropathy, and macroalbuminuria. The prevalence of ASB was 29% in women with type 2 diabetes. Risk factors for ASB in type 2 diabetic women included age, macroalbuminuria, a lower BMI, and a UTI during the previous year. No association was evident between current HbA1c level and the presence of ASB. CONCLUSIONS: The prevalence of ASB is increased in women with diabetes and might be added to the list of diabetic complications in these women. PMID- 10840990 TI - Diabetic neuropathy examination: a hierarchical scoring system to diagnose distal polyneuropathy in diabetes. AB - OBJECTIVE: Existing physical examination scoring systems for distal diabetic polyneuropathy (PNP) do not fulfill all of the following criteria: validity, manageability, predictive value, and hierarchy The aim of this study was to adapt the Neuropathy Disability Score (NDS) to diagnose PNP in diabetes so that it fulfills these criteria. RESEARCH DESIGN AND METHODS: A total of 73 patients with diabetes were examined with the NDS. Monofilaments and biothesiometry were used as clinical standards for PNP to modify the NDS. RESULTS: A total of 43 men and 30 women were studied; mean duration of diabetes was 15 years (1-43), and mean age was 57 years (19-90). A total of 24 patients had type 1 diabetes, and 49 patients had type 2 diabetes. Clinically relevant items were selected from the original 35 NDS items (specific item scored positive in >3 patients). The resulting 8-item Diabetic Neuropathy Examination (DNE) score could accurately predict the results of the clinical standards and is strongly hierarchical (H value 0.53). The sensitivity and specificity of the DNE at a cut-off level of 3 to 4 were 0.96 and 0.51 for abnormal monofilament scores, respectively. For abnormal vibration perception threshold scores, these values were 0.97 and 0.59, respectively. Reproducibility as assessed by inter- and intrarater agreement was good. CONCLUSIONS: The DNE is a sensitive and well-validated hierarchical scoring system that is fast and easy to perform in clinical practice. PMID- 10840991 TI - Health care and health status and outcomes for patients with type 2 diabetes. AB - OBJECTIVE: To evaluate access and utilization of medical care, and health status and outcomes that would be influenced by recent medical care, in a representative sample of patients with type 2 diabetes. RESEARCH DESIGN AND METHODS: A national sample of 733 adults with type 2 diabetes was studied from 1991 to 1994 in the Third National Health and Nutrition Examination Survey. Structured questionnaires and clinical and laboratory assessments were used to determine the frequencies of physician visits, health insurance coverage, screening for diabetes complications, treatment for hyperglycemia, hypertension, and dyslipidemia; and the proportion of patients who met treatment goals and established criteria for health outcome measures including hyperglycemia, albuminuria, obesity, hypertension, and dyslipidemia. RESULTS: Almost all patients had 1 source of primary care (95%), 2 or more physician visits during the past year (88%), and health insurance coverage (91%). Most (76%) were treated with insulin or oral agents for their diabetes, and 45% of those patients taking insulin monitored their blood glucose at least once per day The patients were frequently screened for retinopathy (52%), hypertension (88%), and dyslipidemia (84%). Of those patients with hypertension, 83% were diagnosed and treated with antihypertensive agents and only 17% were undiagnosed or untreated; most of the patients known to have dyslipidemia were treated with medication or diet (89%). Health status and outcomes were less than optimal: 58% had HbA1c >7.0, 45% had BMI >30, 28% had microalbuminuria, and 8% had clinical proteinuria. Of those patients known to have hypertension and dyslipidemia, 60% were not controlled to accepted levels. In addition, 22% of patients smoked cigarettes, 26% had to be hospitalized during the previous year, and 42% assessed their health status as fair or poor. CONCLUSIONS: Rates of health care access and utilization, screening for diabetes complications, and treatment of hyperglycemia, hypertension, and dyslipidemia in type 2 diabetes are high; however, health status and outcomes are unsatisfactory. There are likely to be multiple reasons for this discordance, including intractability of diabetes to current therapies, patient self-care practices, physician medical care practices, and characteristics of U.S. health care systems. PMID- 10840992 TI - Capture-recapture method in the epidemiology of type 2 diabetes: a contribution from the Verona Diabetes Study. AB - OBJECTIVE: The present investigation used data from the Verona Diabetes Study to verify a main assumption of the capture-recapture method (source independence) and to characterize the subgroup of known diabetic patients missed by all sources whose number is estimated by the capture-recapture method. RESEARCH DESIGN AND METHODS: The Verona Diabetes Study identified 7,148 type 2 diabetic patients on 31 December 1986 using 3 different sources: family physicians, a diabetes center, and a drug prescription database. Completeness of ascertainment was estimated with traditional methods based on the hypergeometric distribution and with a log linear model. RESULTS: Identification sources were not independent because the drug prescription database was positively related to family physicians and negatively related to the diabetes center (P < 0.001). Thus, completeness of ascertainment was overestimated (87.5% [95% CI 86.3-88.8]) when using only family physicians and the drug prescription database and underestimated (45.9% [43.9 48.1]) when using only the diabetes center and the drug prescription database. Because of characteristics contributing to variable "catchability" (probability of ascertainment), the estimated proportion of ascertainment increased with increasing time since diagnosis from 65.6% in the first tertile (<6 years) to 91.5% in the third tertile (>12 years); moreover, the ascertainment was estimated to be nearly complete (97.9%) for insulin-treated patients and scanty (28.9%) for diet-treated patients. CONCLUSIONS: Because identification sources are likely to be dependent, the capture-recapture method should be used with caution in diabetes epidemiology and possibly when at least 3 sources are available. The subgroup of diabetic patients whose existence is inferred by this technique likely consists of newly diagnosed patients with mild disease severity. PMID- 10840993 TI - Diabetes Fear of Injecting and Self-Testing Questionnaire: a psychometric evaluation. AB - OBJECTIVE: To study the psychometric properties of the Diabetes Fear of Injecting and Self-Testing Questionnaire (D-FISQ). RESEARCH DESIGN AND METHODS: Two groups of patients were studied. Sample A consisted of 252 insulin-treated diabetes patients. Sample B incorporated 24 insulin-treated patients with high scores (> or = 95th percentile) on the D-FISQ. Test-retest correlations were assessed in both samples. Discriminant and convergent validity of the D-FISQ were assessed with questionnaires concerning fear of hypoglycemia, trait anxiety, and fear of bodily injury, illness, or death. To evaluate criterion-related validity, sample B participated in a behavioral avoidance test (BAT), in which the current level of avoidance of either self-injecting or self-testing was determined. Exploratory factor analysis (EFA) was performed to study whether 2 factors (fear of self injecting [FSI] and fear of self-testing [FST]) could be detected. RESULTS: Test retest correlations ranged from 0.50 to 0.68 (P < 0.001). Correlations between D FISQ and fear of hypoglycemia, trait anxiety, and fear of bodily injury, illness, or death ranged from 0.28 to 0.45 (P < 0.001). Patients who refused to do a BAT for self-injecting or self-testing had higher scores on FSI (P = 0.095) and FST (P = 0.01). EFA yielded 2 separate factors, FSI and FST. CONCLUSIONS: Results from this study support reliability and validity of the D-FISQ, a self-report instrument that can be used for both clinical and research purposes. PMID- 10840994 TI - Impact of type 1 and type 2 diabetes on patterns and costs of drug prescribing: a population-based study. AB - OBJECTIVE: Utilization and costs of prescription drugs were investigated in diabetic and nondiabetic patients. RESEARCH DESIGN AND METHODS: The study was carried out in Tayside, Scotland, U.K. A validated population-based diabetes register was used to identify patients with type 1 and type 2 diabetes, and a database of all prescriptions dispensed in the community was used to investigate drug utilization in 1995. RESULTS: In a population of 406,526, there were 974 (0.2%) with type 1 diabetes and 6,869 (1.7%) with type 2 diabetes. The mean dispensed prescribing rates for all drugs (excluding antidiabetic medication) were higher across all age-groups for diabetic patients. After adjusting for age, patients with type 1 diabetes were 2.07 times (95% CI 2.03-2.11) more likely and patients with type 2 diabetes were 1.70 times (1.69-1.71) more likely to be dispensed a drug item than people without diabetes. This likelihood was increased in every drug category, even those not directly related to diabetes, and the proportion and cost of drug items dispensed to diabetic patients was therefore higher than expected given the prevalence of diabetes. Upon projecting these results to the U.K. population, it was discovered that nearly 8% of the U.K. drug budget (Pound Sterling 350 million) is accounted for by patients with diabetes (90% of that by patients with type 2 diabetes). CONCLUSIONS: This study highlights the increased usage and cost of prescription drugs in diabetes, with type 2 diabetes constituting a particular burden. It was discovered that 1.4% of drug usage in the entire population can be accounted for by the increased prescribing rate of diabetic patients compared with that of nondiabetic patients. PMID- 10840995 TI - Clinical utility of HNF1A genotyping for diabetes in aboriginal Canadians. AB - OBJECTIVE: To determine the diagnostic performance characteristics of HNF1A genotyping for diabetes and impaired glucose tolerance (IGT) in Canadian Oji-Cree Indians. RESEARCH DESIGN AND METHODS: We studied all Oji-Cree subjects > or = 50 years of age (96 subjects) who had participated in a community-wide prevalence survey for type 2 diabetes. Subjects were classified either as having "disease," which included type 2 diabetes and IGT, or not. All subjects were genotyped for the HNF1A G319S mutation. RESULTS: The prevalence of disease in this group was 65.7%, of whom 71.4% had type 2 diabetes. For a carrier of HNF1A S319, the specificity, sensitivity, and positive and negative predictive values were 97.0, 30.1, 95.0, and 42.1%, respectively. When the pretest disease prevalence was accounted for, the probability of disease after a positive test was 97.2%, and the probability of disease after a negative test was 42.2%. The values were very similar for the subgroup of subjects with type 2 diabetes alone. CONCLUSIONS: The HNF1A genotype appears to be the most specific genetic test yet reported for the prediction of a common multifactorial disease by applying present-day standards of clinical epidemiology in molecular genetics. A positive test result had particular diagnostic value in the Oji-Cree: a subject with HNF1A S319 was virtually certain of having diabetes or IGT by 50 years of age. In contrast, a subject without HNF1A S319 had a reduced risk compared with the age-specific prevalence but was not totally risk-free. Because HNF1A S319 was not the only predisposing factor for diabetes in the Oji-Cree, subjects without HNF1A S319 were still at some risk for diabetes or IGT. PMID- 10840996 TI - Insulin action and insulinemia are closely related to the fasting complement C3, but not acylation stimulating protein concentration. AB - OBJECTIVE: An elevated C3 concentration has been reported in people with obesity, type 2 diabetes, hypertension, and dyslipidemia, and has been proposed to play a role in the development of atherosclerosis. We hypothesized that an elevated C3 concentration might be linked to insulin resistance and/or hyperinsulinemia, abnormalities commonly observed in association with the above conditions. RESEARCH DESIGN AND METHODS: Fasting concentrations of C3 and acylation stimulating protein (ASP, C3adesarg), a cleavage product of C3 recently found to stimulate glucose uptake in vitro, were measured in 33 healthy nondiabetic Pima Indians (14 women and 19 men; age 27 +/- 1 and body fat 33 +/- 1%, means +/- SEM). Subjects were characterized for body composition dual-energy X-ray absorptiometry, insulin action (insulin-stimulated glucose disposal [M], hyperinsulinemic glucose clamp), and glucose tolerance (75-g oral glucose tolerance test). RESULTS: Fasting C3 and ASP concentrations were positively correlated (r = 0.43, P < 0.05). Fasting C3 concentration was closely related to percent body fat (r = 0.77), M (r = -0.75), and fasting insulin concentration (r = 0.72) (all P < 0.0001). Fasting C3 concentrations remained significantly related to M and fasting insulin after adjusting for percent body fat (partial r = -0.53 and 0.33, both P < 0.05). In subjects with impaired glucose tolerance, fasting C3 concentrations were higher than in those with normal glucose tolerance -a difference that remained after adjustment for percent body fat and M. We found that fasting ASP concentrations were significantly related to percent body fat (r = 0.37, P < 0.05), but not to M or fasting insulin. CONCLUSIONS: In Pima Indians, fasting C3 concentration is closely related to adiposity, insulin action, and fasting insulin levels and may thus be a mediator for the postulated link between obesity, insulin resistance, hyperinsulinemia, and possibly atherosclerosis. PMID- 10840997 TI - Association of HELLP syndrome with autoimmune antibodies and glucose intolerance. AB - OBJECTIVE: HELLP syndrome is a severe form of preeclampsia, characterized by hemolysis (H), elevated liver enzymes (EL), and low platelets (LP), whose pathogenesis is unclear. Autoimmunity is thought to play an important role. After the observation of development of type 1 diabetes in a patient with HELLP syndrome, we assumed a possible disease association based on autoimmune reactions. RESEARCH DESIGN AND METHODS: We examined 70 women with HELLP syndrome for the presence of autoimmune markers and glucose intolerance. Free thyroxine, triiodothyronine, thyroid-stimulating hormone, anti-thyroglobulin antibodies, thyroperoxidase antibodies, thyrotropin receptor antibodies, antinuclear antibodies (ANAs) and anti-DNA, islet cell antibodies, GADA, an oral glucose tolerance test, and HbA1c were determined postpartum. Patients with positive autoimmune markers or glucose intolerance were prospectively followed and repeated testing was performed. There were 60 women with a normal course of pregnancy matched for age, BMI, and number of pregnancies, which served as a control group. RESULTS: From the HELLP patients, 22 (31%) compared with only 6 (10%) control subjects had autoimmune antibodies (P < 0.01). There were 16 HELLP patients (23%) who exhibited only 1 kind of autoantibody (5 ANA, 9 thyroid antibodies, and 2 GADA), whereas in 6 HELLP patients (8.5%) 2 different antibodies were found. In all but 4 patients of the study group, these antibodies disappeared during 3 +/- 1.5 years of follow-up. Glucose intolerance was detected in 22 (31%) of the HELLP patients, 17 of them had impaired glucose tolerance (IGT), and 5 had diabetes, whereas only 4 subjects (6.5%) with IGT at postpartum were found in the control group (P < 0.01). During the follow-up, 2 HELLP patients were still diabetic and another 2 HELLP patients (1 GADA positive) had IGT versus 1 control subject. CONCLUSIONS: Our data give evidence that HELLP syndrome is associated with various autoimmune antibodies and glucose intolerance. Because glucose intolerance and/or autoimmune markers persisted during long-term follow-up in 6 patients with HELLP syndrome versus 1 in the control group, it may become advisable to reexamine patients with HELLP syndrome for detection of diabetes and autoimmune disorders. PMID- 10840998 TI - MspI polymorphism at +83 bp in intron 1 of the human apolipoprotein A1 gene is associated with elevated levels of HDL cholesterol and apolipoprotein A1 in nondiabetic subjects but not in type 2 diabetic patients with coronary heart disease. AB - OBJECTIVE: Elevated HDL cholesterol and its principal carrier protein apolipoprotein a1 [apo(a1)] are associated with reduced risk of coronary heart disease (CHD). No studies are available on the impact of the -75-bp and/or +83-bp polymorphisms of the apo(a1) gene on HDL cholesterol and apo(a1) levels in patients with type 2 diabetes. RESEARCH DESIGN AND METHODS: We determined the prevalence of the: -75-bp and +83-bp polymorphisms of the apo(a1) gene by restriction fragment length polymorphism analysis among 308 unrelated nondiabetic subjects with CHD and among 251 unrelated patients with type 2 diabetes with CHD and in randomly selected 82 healthy men (CHD-). RESULTS: The rare M1- and M2- allele frequencies of the apo(a1) gene were 23 and 1.8%, respectively, among control subjects; 20 and 1.5%, respectively, among nondiabetic subjects with CHD; and 22 and 2.6%, respectively, among patients with type 2 diabetes and CHD (NS). Nonsmoking nondiabetic subjects with CHD having the M2+- genotype had higher HDL cholesterol (1.48 +/- 0.19 vs. 1.23 +/- 0.02 mmol/l, P < 0.01) and apo(a1) (1.43 +/- 0.10 vs. 1.36 +/- 0.02 g/l, P < 0.05) levels than subjects with the M2++ genotype, even after adjustment for confounding factors. This association was not found among patients with type 2 diabetes and CHD. CONCLUSIONS: We conclude that the +83-bp polymorphism of the apo(a1) gene is associated with elevated HDL cholesterol and apo(a1) levels in Finnish nondiabetic subjects but not in patients with type 2 diabetes. PMID- 10840999 TI - Long-term follow-up in diabetic Charcot feet with spontaneous onset. AB - OBJECTIVE: To assess the long-term results after Charcot breakdown with spontaneous onset in diabetic feet. RESEARCH DESIGN AND METHODS: This study was retrospective. A total of 115 patients (140 feet), 107 with acute deformity and 8 with chronic Charcot deformity, were followed for a median of 48 months (range 6 114). The routine treatment for acute cases was a weight-off regimen with crutches and foot protection with therapeutic shoes until skin temperature had normalized followed by increased weightbearing and the use of bespoke shoes or modification of conventional shoes. RESULTS: The incidence of Charcot deformity was 0.3%/year in the diabetic population investigated. About half of the patients were active in their jobs. Major complications were encountered in 5 (4%) of the patients that required surgical intervention: arthrodesis for unstable malaligned ankles in 3 subjects (1 bilaterally) and major amputation in 2 subjects for unstable ankle and pressure sores. Minor complications were recorded in 43% of subjects: new attacks of Charcot breakdown in 41 patients (36%) and/or foot ulceration in 43 patients (37%) that required minor surgical procedures for 11 patients. All healed except in 2 patients: 1 patient died before the Charcot fractures had healed, and 1 patient died with an unhealed ulcer. No patient lost the ability to walk independently. CONCLUSIONS: Major surgical procedures in only 4% were particularly related to patients with Charcot deformities in the ankle. Minor complications were recorded in about half of the patients. Lifelong foot care is required for diabetic patients with Charcot feet. PMID- 10841000 TI - Effect of menopausal status on insulin-stimulated glucose disposal: comparison of middle-aged premenopausal and early postmenopausal women. AB - OBJECTIVE: Studies in animal models suggest that ovarian hormone deficiency is associated with the development of insulin resistance. In women, ovarian hormone levels are dramatically reduced after the menopause transition. However, the effect of the menopause transition on insulin sensitivity is unclear. Thus, we examined the effect of menopausal status on insulin sensitivity. RESEARCH DESIGN AND METHODS: Insulin-stimulated glucose disposal was measured in 43 middle-aged premenopausal women (47 +/- 3 years of age) during the luteal phase of the menstrual cycle and 40 early postmenopausal women (51 +/- 4 years; time since menopause, 21 +/- 13 months) using the hyperinsulinemic-euglycemic clamp technique. Body composition was measured by dual-energy X-ray absorptiometry and abdominal fat distribution by computed tomography RESULTS: No difference in fat free mass (FFM) was found between groups. Total body (P < 0.01), subcutaneous abdominal (P < 0.05), and intra-abdominal (P < 0.01) adiposity were greater in postmenopausal women compared with premenopausal women. No differences in insulin stimulated glucose disposal were found between premenopausal and postmenopausal women on an absolute basis (pre, 436 +/- 130 vs. post, 446 +/- 120 mg/min), when expressed relative to FFM (pre, 10.7 +/- 3.0 vs. post, 11.5 +/- 3.6 mg x kg(-1) FFM x min(-1)) or when statistically adjusted for FFM (pre, 436 +/- 125 vs. post, 445 +/- 126 mg/min). CONCLUSIONS: These results suggest that menopausal status does not affect insulin sensitivity, as measured by the hyperinsulinemic euglycemic clamp technique. PMID- 10841001 TI - Evaluation of beta-cell secretory capacity using glucagon-like peptide 1. AB - OBJECTIVE: Beta-cell secretory capacity is often evaluated with a glucagon test or a meal test. However, glucagon-like peptide 1 (GLP-1) is the most insulinotropic hormone known, and the effect is preserved in type 2 diabetic patients. RESEARCH DESIGN AND METHODS: We first compared the effects of intravenous bolus injections of 2.5, 5, 15, and 25 nmol GLP-1 with glucagon (1 mg intravenous) and a standard meal (566 kcal) in 6 type 2 diabetic patients and 6 matched control subjects. Next, we studied another 6 patients and 6 control subjects and, in addition to the above procedure, performed a combined glucose plus GLP-1 stimulation, where plasma glucose was increased to 15 mmol/l before injection of 2.5 nmol GLP-1. Finally, we compared the insulin response to glucose plus GLP-1 stimulation with that observed during a hyperglycemic arginine clamp (30 mmol/l) in 8 patients and 8 control subjects. RESULTS: Peak insulin and C peptide concentrations were similar after the meal, after 2.5 nmol GLP-1, and after glucagon. Side effects were less with GLP-1 than with glucagon. Peak insulin and C-peptide concentrations were as follows (C-peptide concentrations are given in parentheses): for patients (n = 12): meal, 277 +/- 42 pmol/l (2,181 +/- 261 pmol/l); GLP-1 (2.5 nmol), 390 +/- 74 pmol/l (2,144 +/- 254 pmol/l); glucagon, 329 +/- 50 pmol/l (1,780 +/- 160 pmol/l); glucose plus GLP-1, 465 +/- 87 pmol/l (2,384 +/- 299 pmol/l); for control subjects (n = 12): meal, 543 +/- 89 pmol/l (2,873 +/- 210 pmol/l); GLP-1, 356 +/- 51 pmol/l (2,001 +/- 130 pmol/l); glucagon, 420 +/- 61 pmol/l (1,995 +/- 99 pmol/l); glucose plus GLP-1, 1,412 +/- 187 pmol/l (4,391 +/- 416 pmol/l). Peak insulin and C-peptide concentrations during the hyperglycemic arginine clamp and during glucose plus GLP-1 injection were as follows: for patients: 475 +/- 141 pmol/l (2,295 +/- 379 pmol/l) and 816 +/- 268 pmol/l (3,043 +/- 508 pmol/l), respectively; for control subjects: 1,403 +/- 308 pmol/l (4,053 +/- 533 pmol/l) and 2,384 +/- 452 pmol/l (6,047 +/- 652 pmol/l), respectively. CONCLUSIONS: GLP-1 (2.5 nmol = 9 microg) elicits similar secretory responses to 1 mg glucagon (but has fewer side effects) and a standard meal. Additional elevation of plasma glucose to 15 mmol/l did not enhance the response further. The incremental response was similar to that elicited by arginine, but hyperglycemia had an additional effect on the response to arginine. PMID- 10841002 TI - Pharmacokinetics of 125I-labeled insulin glargine (HOE 901) in healthy men: comparison with NPH insulin and the influence of different subcutaneous injection sites. AB - OBJECTIVE: To determine the subcutaneous absorption rates and the appearance in plasma of 3 formulations of the long-acting human insulin analog insulin glargine (HOE 901) differing only in zinc content (15, 30, and 80 microg/ml). RESEARCH DESIGN AND METHODS: We conducted 2 studies. Study 1 compared the subcutaneous abdominal injection of 0.15 U/kg of 125I-labeled insulin glargine[15], insulin glargine[80], NPH insulin, and placebo. In study 2, 0.2 U/kg of insulin glargine[30] was injected into the arm, leg, and abdominal regions. Both studies had a randomized crossover design; each enrolled 12 healthy men, aged 18-50 years. RESULTS: In study 1, the time in hours for 25% of the administered radioactivity to disappear after bolus subcutaneous injection (T75%) for NPH insulin indicated a significantly faster absorption rate compared with the 2 insulin glargine formulations (3.2 vs. 8.8 and 11.0 h, respectively P < 0.0001). Mean residual radioactivity with NPH insulin was also significantly lower at 24 h (21.9 vs. 43.8 and 52.2%, P < 0.0001). The calculated plasma exogenous insulin concentrations after NPH insulin were substantially higher than those with insulin glargine, reaching a peak within the first 6 h after administration before declining. Insulin glargine, however, did not exhibit a distinct peak. Weighted average plasma glucose concentration between 0 and 6 h was significantly lower after NPH compared with insulin glargine (P < 0.001). In study 2, there were no significant differences in the absorption characteristics of insulin glargine between the 3 injection sites (T75% = 11.9, 15.3, and 13.2 h for arm, leg, and abdomen, respectively) or in residual radioactivity at 24 h. CONCLUSIONS: Subcutaneous absorption of insulin glargine is delayed compared with NPH insulin. There is little or no difference in the absorption rate of insulin glargine between the main subcutaneous injection sites. PMID- 10841003 TI - Diabetes in urban African-Americans. XIX. Prediction of the need for pharmacological therapy. AB - OBJECTIVE: To develop a prediction rule that will identify patients who will require pharmacological therapy within 6 months of first presentation to a diabetes clinic. RESEARCH DESIGN AND METHODS: Among the patients who came to the Grady Diabetes Clinic between 1991 and 1997, we randomized 557 frequent attenders to a development group and 520 frequent attenders to a validation group. Using multiple logistical regression, we derived a prediction rule in the development group to project whether patients would require pharmacological intervention to achieve HbA1c levels <7% after 6 months. The utility of the prediction rule was then confirmed in the validation group and tested prospectively on an additional group of 93 patients who presented from 1997 to 1998. Performance of the prediction rule was assessed using receiver operating characteristic (ROC) curves. RESULTS: The rule (-4.469 + 1.932 x sulfonylurea Rx + 1.334 x insulin Rx + 0.196 x duration + 0.468 x fasting glucose, where "Rx" indicates a prescription) predicted the need for pharmacological intervention in the development group (P < 0.0001). Use of insulin or sulfonylurea therapy at presentation, duration of diabetes, and fasting glucose levels were significant predictors of the future need for pharmacological management. The prediction rule also performed well in the validation group (positive predictive value 90%, correlation between predicted and observed need for medical management 0.99). ROC curves confirmed the value of the prediction rule (area under the curves was 0.91 for the development group, 0.85 for the validation group, and 0.81 for the prospective group). CONCLUSIONS: Early identification of individuals who will require pharmacological intervention to achieve national standards for glycemic control can be achieved with high probability, thus allowing for more efficient management of diabetes. PMID- 10841004 TI - Is there a glycemic threshold for impaired autonomic control? AB - OBJECTIVE: Although hyperglycemia has been recognized as a predictor of cardiovascular autonomic neuropathy in diabetic patients, the glucose threshold at which autonomic control begins to become impaired has not been evaluated. This study examined whether fasting plasma glucose (FPG) or fasting plasma insulin (FPI) is associated with reductions in baroreflex sensitivity (BRS) in healthy volunteers. RESEARCH DESIGN AND METHODS: FPG and FPI were measured after an overnight fast in 162 healthy volunteers (91 men, 71 women) who were 25-44 years of age. BRS was measured with power spectral analysis. RESULTS: Univariate analyses showed that FPG was negatively correlated with BRS (r = -0.25, P < or = 0.001) with significant reductions observed in volunteers with FPG in the upper 2 quintiles (i.e., 93-124 mg/dl). However, after adjustment for other predictors of BRS (e.g., age, blood pressure, and BMI), the relationship between FPG and BRS was no longer significant. In contrast, FPI was negatively correlated with BRS in univariate analyses (r = -0.32, P < 0.0001) as well as after covariate adjustment, with close to a 50% reduction in BRS observed in the volunteers with insulin values in the highest quintile (i.e., 16-36 microU/ml). CONCLUSIONS: These findings suggest that high normal levels of FPG are associated with reduced autonomic control secondary to the effects of aging, obesity, and elevated blood pressure on FPG levels and that elevations in FPI are associated with substantial reductions in autonomic cardiac control independent of other covariates. PMID- 10841005 TI - Polymorphism of the tumor necrosis factor-alpha receptor 2 gene is associated with obesity, leptin levels, and insulin resistance in young subjects and diet treated type 2 diabetic patients. AB - OBJECTIVE: Mice lacking the tumor necrosis factor-alpha receptor 2 (TNFR2) gene fed a high-fat diet gain less weight and display reduced leptin and insulin levels. In humans, plasma levels of the soluble fraction of TNFR2 (sTNFR2) circulate in proportion to the degree of insulin resistance. The purpose of this study was to evaluate a polymorphism in the 3' untranslated region of the TNFR2 gene on chromosome 1 in relation to BMI, leptin levels, and insulin resistance. RESEARCH DESIGN AND METHODS: Using single-strand conformation polymorphism, the polymorphism was analyzed in 107 nondiabetic subjects (60 women, 47 men) and in 110 consecutive patients with type 2 diabetes (79 women, 31 men). In a subset of 33 healthy subjects, insulin sensitivity (minimal model analysis) was also evaluated. RESULTS: Four alleles of the TNFR2 gene were identified (A1, A2, A3, and A4). BMI and serum leptin levels were significantly increased in young carriers of the A2 allele. Plasma sTNFR2 levels were similar among the different TNFR2 gene variants. However, in subjects who did not carry the A2 allele, in young subjects, and in women, plasma sTNFR2 levels were proportional to BMI and leptin levels. In the study sample, carriers of the A2 allele (n = 18) showed significantly increased BMI, fat mass, waist-to-hip ratio, serum total and VLDL triglyceride levels, and leptin levels and had a lower insulin sensitivity index than noncarriers of the A2 variant (n = 15). The frequency of the different alleles among diabetic subjects was similar to that in the control population. However, diet-treated diabetic subjects (n = 49) who were carriers of the A2 allele exhibited significantly higher BMI and leptin levels than diet-treated noncarriers of the A2 allele. CONCLUSIONS: The presence of the A2 allele in the TNFR2 gene may predispose subjects to obesity and higher leptin levels, which may in turn predispose them to insulin resistance or vice versa. The TNFR2 gene may be involved in weight-control mechanisms. PMID- 10841006 TI - High frequency of persisting or increasing islet-specific autoantibody levels after diagnosis of type 1 diabetes presenting before 40 years of age. The Belgian Diabetes Registry. AB - OBJECTIVE: To study the presence and levels of GAD65 antibodies (GADA), IA-2 antibodies (IA-2-A), and islet cell antibodies (ICA) during the first years after clinical onset of type 1 diabetes in relation to age at diagnosis. RESEARCH DESIGN AND METHODS: Type 1 diabetic patients (n = 194) <40 years of age were consecutively recruited at the time of diagnosis by the Belgian Diabetes Registry and followed during the first 4 years of insulin treatment. ICA were determined by indirect immunofluorescence assay and IA-2-A, GADA, and insulin autoantibodies by a radioligand assay. RESULTS: Overall, 94% of initially antibody-positive patients (n = 180) remained positive for at least 1 antibody type 4 years after diagnosis. In the case of diagnosis after 7 years of age, GADA, IA-2-A, and ICA persisted in 91, 88, and 71%, respectively, of the initially antibody-positive patients. Antibody persistence was lower in those diagnosed at <7 years of age, amounting to 60% for GADA, 71% for IA-2-A, and 39% for ICA. In 57% of the initially antibody-positive patients, at least 1 type of autoantibody reached peak values after diagnosis. This occurred more frequently for clinical onset after 7 years of age and more often for GADA (49%) than for IA-2-A (29%) or ICA (19%). Of the patients, 24% that were negative for GADA at onset became GADA positive during the following 4 years. Among the 7% initially antibody-negative patients, 2 of 14 subjects developed antibodies after clinical onset. CONCLUSIONS: In particular, for diagnosis after 7 years of age, islet cell specific autoantibodies generally persist for many years after diagnosis. There is also a high frequency of increasing antibody levels and of conversion to antibody positivity in the first 4 years after diagnosis and start of insulin treatment. Thus, determination of antibodies at diagnosis can underestimate the number of cases with autoimmune type 1 diabetes, in particular with assays of lower sensitivity. The divergent temporal patterns of ICA, GADA, and IA-2-A suggest that the ICA test recognizes other antibody specificities besides GADA and IA-2-A and reflects other autoimmune processes; it also indicates that GADA assays have a higher diagnostic sensitivity in the period after clinical onset. PMID- 10841007 TI - American Diabetes Association Annual Meeting, 1999: more on cardiovascular disease. PMID- 10841008 TI - Healthy people 2010: diabetes. PMID- 10841009 TI - Role of triglyceride levels in identifying insulin resistance in nonobese type 2 diabetic Japanese patients with hypertension. PMID- 10841010 TI - Effect of physical training on insulin sensitivity in Japanese type 2 diabetic patients: role of serum triglyceride levels. PMID- 10841011 TI - Plasma brain natriuretic peptide levels in normotensive type 2 diabetic patients without cardiac disease. PMID- 10841012 TI - Breath hydrogen testing identifies patients with diabetic gastroparesis. PMID- 10841013 TI - Insulin-induced hypoglycemia induces a rise in C-reactive protein. PMID- 10841014 TI - Trp64Arg polymorphism of the beta3-adrenergic receptor is not associated with diabetic nephropathy in Japanese patients with type 2 diabetes. PMID- 10841015 TI - Telemedicine in the management of pregnancy in type 1 diabetic women. PMID- 10841016 TI - Gly82Ser polymorphism of the receptor of advanced glycation end product gene is not associated with coronary heart disease in Finnish nondiabetic subjects or in patients with type 2 diabetes. PMID- 10841017 TI - Relationship between plasma adrenomedullin levels and metabolic control, risk factors, and diabetic microangiopathy in patients with type 2 diabetes. PMID- 10841018 TI - Cautionary note regarding HbA1c methods predicting the clinical status of diabetic patients. PMID- 10841019 TI - Where is the evidence that radial artery tonometry can be used to accurately and noninvasively predict central aortic blood pressure in patients with diabetes? PMID- 10841020 TI - Hospital management of diabetic ketoacidosis in the U.K. PMID- 10841021 TI - Hemophilus vaccine associated with increased risk of diabetes: causality likely. PMID- 10841022 TI - Consensus Development Conference on diabetic foot wound care: a randomized controlled trial does exist supporting use of adjunctive hyperbaric oxygen therapy. PMID- 10841023 TI - Response to Bazzigaluppi et al.: capillary whole-blood measurement of islet autoantibodies. PMID- 10841024 TI - HbA1c determination with high-performance liquid chromatography. PMID- 10841025 TI - The 12-Item Well-Being Questionnaire: origins, current stage of development, and availability. PMID- 10841026 TI - Cyclic AMP a key messenger in the regulation of skin pigmentation. AB - Compelling evidence has been gathered indicating that pro-opiomelanocortin peptides, alpha-melanocyte stimulating hormone (alpha-MSH) and adrenocorticotropic hormone (ACTH), through the cyclic AMP pathway, play a pivotal role in melanocyte differentiation and in the regulation of melanogenesis. Recently, the molecular events linking cAMP to melanogenesis up regulation have been elucidated. This cascade involves the activation of protein kinase A and CREB transcription factor, leading to the up-regulation of the expression of Microphthalmia associated transcription factor (MITF). MITF has been found mutated in patients with Waardenburg syndrome 2A, and plays a crucial role in melanocyte development. MITF binds and activates melanogenic gene promoters, thereby increasing their expression which results in an increased melanin synthesis. Beyond this simplified scheme, It appears that melanogenic gene expression is controlled by a complex network of regulation involving other transcription factors such as Brn2, TBX2, PAX3 and SOX10. Further studies are required to better understand the respective roles of these factors in the regulation of melanin synthesis. In addition, other intracellular signaling pathways, like the phosphatidyl inositol 3-kinase pathway, as well as the molecular cascade of events governed by the small GTP-binding protein Rho, seem to be involved in the regulation of melanogenesis and melanocyte dendricity. Finally, it should be mentioned that cAMP activates a melanocyte-specific pathway leading to MAP kinase activation. MAP kinase, ERK2, phosphorylates MITF, thereby targeting the transcription factor to proteasomes for degradation. Thus, in addition to the complex transcriptional regulation, melanogenesis is also subjected to a post-translational regulation that controls MITF or tyrosinase function. Taken together, these complex molecular processes would finally allow a fine tuning of melanocyte differentiation leading to melanin synthesis. PMID- 10841027 TI - Nomenclature for identified pigmentation genes in the mouse. AB - More than 90 different loci influence pigmentation in the mouse. During the past few years, an increasing number of genes have been identified, and assigned to the corresponding coat color loci and pigmentation mutants. As a consequence, different names have been used in publications for loci, genes and corresponding proteins. In the following article, we present the rules and guidelines for gene nomenclature, and provide the current nomenclature for pigmentation mutants in the mouse. PMID- 10841028 TI - Gene correction by RNA-DNA oligonucleotides. AB - An oligonucleotide composed of a contiguous stretch of RNA and DNA residues has been developed to facilitate the correction of single-base mutations of episomal and chromosomal targets in mammalian cells. The design of the oligonucleotide exploited the highly recombinogenic RNA-DNA hybrids and featured hairpin capped ends avoiding destruction by cellular helicases or exonucleases. The RNA-DNA oligonucleotide (RDO) was designed to correct a point mutation in the tyrosinase gene and caused a permanent gene correction in mouse albino melanocytes, determined by clonal analysis at the level of genomic sequence, protein and phenotypic change. Recently, we demonstrated correction of the tyrosinase gene using the same RDO in vivo, as detected by dark pigmentation of several hairs and DOPA staining of hair follicles in the treated skin of albino mice. Such RDOs might hold a promise as a therapeutic method for the treatment of skin diseases. However, the frequency of gene correction varies among different cells, indicating that cellular activities, such as recombination and repair, may be important for gene conversion by RDOs. As this technology becomes more widely utilized in the scientific community, it will be important to understand the mechanism and to optimize the design of RDOs to improve their efficiency and general applicability. PMID- 10841029 TI - Melanin granules prevent the cytotoxic effects of L-DOPA on retinal pigment epithelial cells in vitro by regulation of NO and superoxide radicals. AB - Inasmuch as the nitrogen cycle elicits the direct reduction of N2 to NH3 through enzymatic reactions, and inasmuch as L-DOPA (L-dihydroxyphentlalamine), a catecholamine, can be a source of nitric oxide (NO), it is possible that melanin granules in the eye affect the generation of NO, which causes damage to the retinal pigment epithelial (RPE) cells during the oxidation of L-DOPA. In order to confirm this possibility, we analyzed the correlations of NO generation, cell growth, and superoxide dismutase (SOD) activities in two types (melanotic and amelanotic) of bovine RPE cells following exposure to L-DOPA. NO generation from L-DOPA was determined using an NO detector that is reliant on redox currents. The concentration of NO was measured in terms of diffusion currents run between a working electrode and a counter electrode, both being set in culture medium placed in a Petri dish. For the assays, L-DOPA was added to the medium at various concentrations (5, 29.9, 79.4, 152.7 or 249 microM), and 6 min after addition, an NO-trapping agent 2,4-carboxyphenyl-4,4,5,5-tetramethylimidazole-1-oxyl 3-oxide (carboxy-PTIO) was also added. The melanotic and amelanotic types of RPE cells were cultured separately in medium with L-DOPA under an atmosphere containing 20, 10 or 5% oxygen. Cell numbers were counted using a Coulter counter, and SOD activities were determined following incubation for 24, 48 or 72 hr using a modification of the luminol assay. The results obtained indicated that: (a) NO was produced from L-DOPA in a concentration-dependent manner and was trapped quantitatively by carboxy-PTIO; (b) the generation of NO was inhibited more markedly in the melanotic cell line than in the amelanotic one, suggesting an increased tolerance to L-DOPA-derived cytotoxicity in the former; and (c) the SOD activities were more affected by oxygen concentration in the melanotic cells than in the amelanotic ones. From these results, it is concluded that melanin granules in RPE cells have a role in preventing the cytotoxicity derived from L-DOPA and in regulating the generation of NO and superoxide radicals. PMID- 10841030 TI - Inhibition of UVR-induced tanning and immunosuppression by topical applications of vitamins C and E to the skin of hairless (hr/hr) mice. AB - Exposure of C3HBYB/Wq hairless (hr/hr) mice to ultra-violet radiation (UVR) for 15 days induced intense tanning of their dorsal skin. Small, dark freckles appeared first, gradually enlarging and coalescing as treatment progressed yielding a uniform tan. Histologically, the gross changes in skin color were matched initially by the appearance of scattered epidermal melanocytes that subsequently proliferated to form discrete, progressively expanding and abutting populations resulting in a uniform melanocyte network throughout the basal layer of the interfollicular epidermis. In contrast, when applied topically before each daily exposure to UVR, a cream or lotion vehicle containing both vitamins C and E (Vits C/E) inhibited UVR-induced erythema and tanning. Application of Vits C/E, both before and after irradiation, was no more effective in providing photoprotection than pre-treatment only. At the tissue level, UVR-induced proliferation and melanogenesis of melanocytes were reduced compared with irradiated controls. The density of individual melanocyte populations was reduced, as was the number of melanocyte populations achieving merger (confluence) with others. Confluence grades and cell counts, estimating the maximum density of melanocyte populations in UVR-Vits C/E-treated mice, were approximately two thirds those of UVR-vehicle-treated controls. However, tanning was only one fifth that of UVR-vehicle-treated controls, suggesting that melanogenesis was also inhibited. In addition to its inhibitory actions on irradiated melanocytes, Vits C/E also inhibited UVR-induced suppression of contact hypersensitivity (CHS) in haired (Hr/hr) and hr/hr mice of the C3HBYB/Wq strain. The common denominators for most, if not all, of the influences of topically-applied Vits C/E in muting the responses of the melanocyte and immune systems to UVR may stem from the vitamins' combined ability to suppress UVR stimulated inflammation and its associated cascade of mediators. PMID- 10841031 TI - Structural differences in unbleached and mildly-bleached synthetic tyrosine derived melanins identified by scanning probe microscopies. AB - Using scanning tunneling microscopy (STM), we have imaged two types of mildly bleached, synthetic tyrosine-derived melanins for comparison with the unbleached melanin from which they were prepared. These mildly-bleached melanins were generated by mild oxidation of the unbleached melanin, using either basic hydrogen peroxide or air/light. The unbleached melanin, and two mildly-bleached melanins, were independently deposited from very dilute tetrahydrofuran (THF) solutions onto highly oriented pyrolytic graphite (HOPG) substrate for STM imaging. Lateral dimensions (23 A, average of two directions) of structures from each of the three samples showed no differences. However, structures from both mildly-bleached melanins showed similar dramatic decreases (from approximately 15 A to approximately 5 A) in their STM-measured apparent heights, compared with structures from the unbleached melanin sample. These STM observations are compatible with structural models for unbleached and mildly-bleached melanins, incorporating a three-dimensional structure for unbleached melanin composed of multi-layered, pi-pi-stacked, carboxylic and amino variants of polyaromatic polymeric sheets. The STM-observed decrease in apparent heights after mild oxidation, which we associate with a change in stack height, has been confirmed by experiments using tapping mode atomic force microscopy (TM-AFM) for the unbleached and mildly-hydrogen-peroXide-bleached melanins (from approximately 14 A to approximately 6 A). In these TM-AFM experiments, the melanins were deposited directly onto magnesium cation-treated glass substrates in contact with methanolic solutions of each of the melanins. We interpret our mild-bleaching results as an oxidative conversion of the multi-layered, stacked sheets of mainly carboxylic and amino variants of polyquinhydrone-like moieties, to largely de stacked, mildly-bleached melanin sheets. These oxidized and, hence, electron deficient sheets should not readily form multi-layered, pi-pi interacting stacks, but instead appear to be either single-layer polyquinone sheets or, at most, double-layer polyquinhydrone sheets. The effects of such de-stacking on in vivo melanin photoprotection, and structural similarities between melanin derived from natural sources and the synthetic melanin samples used in this work are discussed. PMID- 10841032 TI - A possible mechanism for feedback regulation of the mouse tyrosinase gene by its 3' non-coding RNA fragments. AB - The 5' upstream regulatory region of the mouse tyrosinase gene contains a long (GA)n sequence, that may be capable of adopting a triple-helical conformation (triplex). We analyzed protein-DNA interactions in a part of the 5' upstream region containing the (GA)n sequence by gel retardation analysis and found evidence for a cell type-specific protein(s) that bound to this region. We also found a (TC)10 sequence about 100 bp downstream from a polyadenylation site of the gene. Examination of tyrosinase cDNAs and Northern analysis indicated that this sequence is transcribed and removed during 3' end-processing of the mRNA. Based on the hypothesis that the (TC)10 sequence binds to the (GA)n sequence and forms an intermolecular triplex, we performed the same gel retardation assay in the presence of the 3' non-coding RNA fragments containing the (UC)10 sequence. The probe DNA failed to interact with the cell type-specific protein(s). These results suggest a novel hypothesis for the regulation of the mouse tyrosinase gene, i.e. that the 3' non-coding RNA fragments of mouse tyrosinase transcripts suppress its own expression at the transcriptional level. This might occur by preventing cell type-specific protein factor(s) from binding to the regulatory cis-elements in the 5' upstream region of the gene, possibly through a triplex formation, although this hypothesis remains to be proven. PMID- 10841033 TI - Microspectrophotometric analysis of intact chromatophores of the Japanese medaka, Oryzias latipes. AB - To investigate the possible photoprotective role of chromatophores in fish, the absorbances of four types of intact chromatophores in adult and larval Japanese medaka were analyzed using microspectrophotometric techniques. The absorbance spectrum of each chromatophore class was obtained from 300 to 550 nm. The absorbance spectra of intact leucophores, melanophores and xanthophores were very similar to the published absorbance spectra of the isolated pure pigments contained in each chromatophore type, pteridines, melanin and carotenoids or pteridines, respectively. Based on these absorbance spectra, leucophores and melanophores should provide the most ultraviolet (UV) photoprotection to fish since the compounds they contain, pteridines and melanin, correspondingly, have strong absorbances in the UV region of the spectrum. Xanthophores containing carotenoids are not likely to provide much protection to fish from UV-induced damage since carotenoids have low absorbances in the UV range. Xanthophores containing colored pteridines, however, may provide somewhat greater UV protection to fish, since pteridines absorb more light than carotenoids in the UV portion of the spectrum. The relative frequency, coverage and thickness of these two types of xanthophores should determine how much protection xanthophores as a chromatophore type would provide against UV-induced damage. PMID- 10841034 TI - Regulation of the murine silver locus product (gp87) by the hypopigmenting cytokines TGF-beta1 and TNF-alpha. AB - The melanosomal proteins encoded by the silver (si, SILV, or PMEL17) locus play important roles in melanogenesis and are actively investigated as targets for melanoma immunotherapy. The human silver locus yields two proteins, gp100 and PMEL17, by alternative splicing of a common mRNA precursor. Mouse melanocytes exclusively express the gp100 orthologue, here termed gp87, thus providing a simpler model with which to study the silver locus products. We have analyzed the effects of [Nle4, D-Phe7]-alpha melanocyte-stimulating hormone (alphaMSH) and two hypopigmenting cytokines, tumor necrosis factor (TNF)-alpha and transforming growth factor (TGF)-beta1, on the expression of gp87 in B16 mouse melanoma cells. TNF-alpha and TGF-beta1 (at saturating doses for 48 hr) decreased gp87 mRNA by 50%. The gp87 protein was almost undetectable by Western immunoblotting after TNF alpha treatment, but was not affected by TGF-beta1. alphaMSH increased the mRNA and the gp87 protein approximately 2-fold. Moreover, the amount of gp87 was not reduced by TNF-alpha in the presence of the hormone, in spite of a 50%, decrease in its mRNA. Therefore, the levels of mRNA and gp87 protein did not correlate after treatment with the cytokines. Overall, our data suggest that the silver locus product is not regulated exclusively at the transcriptional level, and highlight the importance of still-uncharacterized regulatory translational and/or post-translational events. PMID- 10841035 TI - The role of prostaglandins in penile erection. AB - The balance of penile smooth muscle tone is finely controlled, with contractile factors acting in opposition to relaxant factors. The principal agents in this process are undoubtedly noradrenaline and nitric oxide. Prostaglandins probably have a crucial role in the 'fine tuning' of corporal smooth muscle tone. Their effects on control mechanisms in the healthy penis are more likely to be modulatory rather than direct. PMID- 10841036 TI - Dietary docosahexaenoic acid but not eicosapentaenoic acid suppresses lipopolysaccharide-induced interleukin-1 beta mRNA induction in mouse spleen leukocytes. AB - Mice were fed a diet supplemented either with beef tallow (BT), BT plus ethyl eicosapentaenoate (EPA) or BT plus ethyl docosahexaenoate (DHA) for 9 weeks. EPA and DHA supplementation increased the content of the respective fatty acid in spleen leukocyte lipids, which was associated with the reduction in the arachidonate content. IL-1beta mRNA induction upon lipopolysaccharide (LPS) stimulation in spleen leukocytes in the DHA diet group was significantly lower than in the BT diet group, but the EPA diet was without any significant effect. The amount of prostaglandin E2 (PGE2) released from LPS-stimulated spleen leukocytes was significantly lower in both the EPA and DHA groups than in the BT group. Thus, dietary EPA and DHA inhibited arachidonate metabolism similarly but had different effects on IL-1beta mRNA induction in mouse spleen leukocytes. PMID- 10841037 TI - The effect of cyclic GMP on rabbit corporal smooth muscle tone and its modulation by cyclo-oxygenase products. AB - Corporal smooth muscle (CSM) tone is maintained by a finite balance between relaxant and contractile neurotransmitters. The aim of these experiments was to ascertain the degree to which cyclic GMP is involved in these interactions. We also sought to elucidate the pharmacological mechanism of action of MB in rabbit corpus cavernosum (RCC), an important tool in nitric oxide research. Using an organ chamber technique, strips of RCC were treated with the guanylate cyclase inhibitors Methylene Blue (MB) and LY83583; 100 microM MB led to increases in resting tension which were antagonized by indomethacin, nifedipine, phentolamine, but not superoxide dismutase (SOD). Contractile responses to noradrenaline (NA) were increased and relaxation to ACh was impaired by both MB and LY83583 and reversed with indomethacin, but not SOD. Pyrogallol had no effect on agonist induced responses. The pharmacological action of MB in RCC does not depend on the generation of superoxide anions. Endothelium-dependent relaxation in RCC results in activation of soluble guanylate cyclase and release of a stable endothelium derived contracting factor(s), which is likely to be a constrictor prostanoid(s). Tonic production of cGMP in RCC inhibits the presynaptic release and contractile effects of NA and can be modulated by cyclo-oxygenase inhibition, demonstrating the important interaction and functional antagonism between cGMP and prostaglandins in the control of CSM tone. PMID- 10841038 TI - Differential effects of nitric oxide on the activity of prostaglandin endoperoxide H synthase-1 and -2 in vascular endothelial cells. AB - A number of studies have demonstrated that prostacyclin and nitric oxide (NO) regulate blood pressure, blood flow and platelet aggregation. In this paper, we have examined the possible relationship between NO and prostaglandin endoperoxide H synthase (PGHS)-1 and -2 activities in cultured bovine aortic endothelial cells. In the non-activated condition endothelial cells expressed PGHS-1 activity alone. When these cells were pretreated with aspirin to inactivate their PGHS-1 and then activated by serum and phorbol ester (TPA) for 6 h, the cells expressed PGHS-2 activity alone. The PGHS activity was assessed by the generation of 6 ketoprostaglandin F1alpha (6-ketoPGF1alpha), a stable metabolite of prostacyclin, after the treatment of these cells with arachidonic acid. The simultaneous addition of NOC-7, a NO donor, with arachidonic acid did not affect the production of 6-ketoPGF1alpha in PGHS-1 expressed cells, but attenuated it in PGHS-2-expressed cells. The inhibitory effect of NOC-7 on PGHS-2 activity was dose dependent, and the different effects of NOC-7 on the activities of PGHS isozymes were also observed in other NO donors. To confirm the different effect of NO on PGHS isozymes demonstrated in the cultured endothelial cells, we carried out an ex vivo perfusion assay in aorta isolated from normal and lipopolysaccharide (LPS)-treated rats. In the aortae isolated from normal rats, where dominant expression of PGHS-1 was expected, the NO donor did not affect the PGHS activity, while in aortae isolated from LPS-treated rats, where PGHS-2 was dominantly expressed, the NO donor dramatically inhibited the PGHS activity, suggesting that NO suppressed PGHS-2 activity alone. The inhibitory effect of NO on PGHS-2 activity was not mediated by cyclic GMP (cGMP), since (a) methylene blue, an inhibitor of soluble guanylate cyclase did not abolish the inhibitory effect of the NO donor on PGHS-2 activity, and (b) 8-Br-cGMP, a permeable cGMP analogue, failed to mimic the effect of NO donors. These data suggest that the effect of NO on prostacyclin production in endothelial cells was dependent on the expression rate of PGHS-1 and PGHS-2 in the cells. PMID- 10841039 TI - Depletion of delta 9 desaturase (EC 1.14.99.5) activity in lactating rat during protein restriction. AB - The effects of protein restriction on the activity of delta9 desaturase (EC 1.14.99.5) were investigated in lactating rats. A control group was fed a balanced diet (20% casein) for 14 days, whereas the experimental groups were fed a low-protein diet (8% casein), supplemented with or without L-methionine (0.4%), for 14 days. The enzyme activity was measured by incubations of hepatic microsomal pellets with (1-14C) stearic acid. Results showed a decreased delta9 desaturase activity, after 2,7 and 14 days of depleted diet, of -50, -40 and -33% respectively, compared with control. The supplementation of the low-protein diet with 0.4% methionine, which favours food consumption as well as growth, did not improve the altered delta9 desaturase activity. Our data evidenced that delta9 desaturase activity is depleted by protein restriction during lactation, when the protein needs are high for the biosynthesis of animal tissues. This change has to be considered as a sign of depressed delta9 desaturase biosynthesis or modifications of enzymatic properties, or both. PMID- 10841040 TI - Determination of free and glucuronide conjugated 20-hydroxyarachidonic acid (20 HETE) in urine by gas chromatography/negative ion chemical ionization mass spectrometry. AB - 20-Hydroxy-arachidonic acid (20-HETE) was determined in urine by an isotope dilution assay using gas chromatography/mass spectrometry (GC/MS). After addition of 18O2-internal standard, 20-HETE was extracted from urine with hexane either directly or after treatment with glucuronidase. 20-HETE was derivatized to the pentafluorobenzylester and the sample was applied to thin layer chromatography with iso-octane/iso-propanol 9:1 (v/v) as the developing solvent. The corresponding zone was extracted and 20-HETE was hydrogenated. After derivatization to the trimethylsilylether, 20-HETE was determined by GC/MS using the [M-pentafluorobenzyl]- -ion in the negative ion chemical ionization mode. Excretion rates of free and glucuronide conjugated 20-HETE was determined in healthy children and in children with hyperprostaglandin-E-syndrome/antenatal Bartter syndrome (HPS/aBS) with or without indomethacin treatment. Compared to the controls, the HPS/aBS children showed higher excretion rates of 20-HETE, which were suppressed to normal values under indomethacin medication. Free and glucuronide conjugated 20-HETE do not correlate with PGE2 excluding any participation in HPS/aBS. PMID- 10841041 TI - Inhibition of duodenal enterocyte Mg2+-ATPase by arachidonic acid is not mediated by an effect on protein kinase C. AB - Active absorption processes in the duodenal enterocyte are driven by various ATPases. It is known that the activity of Na+,K+-ATPase, Ca2+-ATPase and Mg2+ ATPase can be modulated by polyunsaturated fatty acids of the n-6 series, for example by linoleic and gamma-linolenic acids. These effects may be achieved by protein phosphorylation via protein kinase C. The present study was undertaken to determine the effect of arachidonic acid on Mg2+-ATPase (measured colorimetrically) activity in basolateral membranes prepared from rat duodenum. It shows, for the first time, significant dose-dependent inhibition of Mg2+ ATPase (26-62%) by arachidonic acid (10-50 microg/ml) which already takes place after one minute of exposure, indicating involvement of a rapid signal transduction mechanism. Addition of the protein kinase C inhibitors bisimidolylmaleimide (2.5 microM) and calphostin (0.5 microM) did not influence the action of arachidonic acid on Mg2+-ATPase; protein kinase C involvement in this process is thus not indicated. PMID- 10841042 TI - Involvement of arachidonic acid in chemical stress-induced interleukin-6 synthesis in osteoblast-like cells: comparison with heat shock protein 27 induction. AB - In a previous study, we have demonstrated that sodium arsenite (arsenite) as chemical stress stimulates heat shock protein 27 (HSP27) induction and arachidonic acid release in osteoblast-like MC3T3-E1 cells, and that the response of HSP27 induction is coupled with metabolic activity of the arachidonic acid cascade. In the present study, we examined the effect of exposure to arsenite on the synthesis of interleukin-6 (IL-6) in these cells. Arsenite induced the synthesis of IL-6 after 6 h from the stimulation up to 48 h. The effect of arsenite on IL-6 synthesis was dose-dependent in the range between 10 and 500 microM. The arsenite-induced IL-6 synthesis was enhanced by the pretreatment with indomethacin, an inhibitor of cyclooxygenase. Nordihydroguaiaretic acid, a lipoxygenase inhibitor, significantly amplified the arsenite-induced IL-6 synthesis. Melittin, an activator of phospholipase A2, which by itself hardly affected the levels of IL-6, markedly enhanced the arsenite-induced IL-6 synthesis. These results strongly suggest that chemical stress induces IL-6 synthesis in osteoblasts, and that the IL-6 synthesis is coupled to the arachidonic acid cascade as well as the HSP27 induction by arsenite. PMID- 10841043 TI - Vitamin E in relation to lipid peroxidation in experimental septic shock. AB - Lipid peroxidation and antioxidant balance in the body is a crucial factor in the pathophysiology of various diseases. This study investigates the circulatory alpha-tocopherol levels and its relationship with 8-iso-prostaglandin F2alpha (8 iso-PGF2alpha), a non-enzymatic and, 15-keto-13,14-dihydro-PGF2alpha (15-K-DH PGF2alpha), a cyclooxygenase catalysed oxidation product of arachidonic acid in experimental septic shock in pigs. A steady decrease in alpha-tocopherol levels in plasma was observed in both survivor and non-survivor animals. A simultaneous increase of oxidative injury indicator, plasma 8-iso-PGF2alpha was seen in both groups but with a different fashion. 8-Iso-PGF2alpha levels increased steadily in the animals that died during the experiment. An early and rapid increase of plasma 15-K-DH-PGF2alpha, an inflammatory response indicator, was also observed in all animals. There was a significant difference in the kinetics of decrement of alpha-tocopherol levels and a concomitant increase in 15-K-DH-PGF2alpha levels among the non-survivors. Thus, a successive disappearance of circulatory vitamin E in conjunction with the surge of plasma isoprostanes and prostaglandins impairs the oxidant-antioxidant balance in favour of the former and may possibly have an effect on the survivality during experimental porcine septicaemia. PMID- 10841044 TI - Effects of n-3 fatty acids on growth and survival of J774 macrophages. AB - To further understand potential mechanisms underlying the protective effects of eicosapentanoic acid (EPA) against atherosclerosis, J774 macrophages were used to explore cellular responses to growth in the presence of PUFA in vitro. Clonogenic assays indicated that 15 microg/ml of EPA killed over 90% of J774 populations. Docosapentaenoic acid (DPA) was more cytotoxic than either EPA or docosahexaenoic acid (DHA). EPA was shown to be elongated to DPA. Cytotoxicity induced by EPA was not inhibited by the presence of alpha-tocopherol (a-toc) in the medium. Immunological screening for caspase enzymes and microscopic examination indicated that apoptosis was not the major cause of cell death. Proliferation assays demonstrated that total cell numbers of EPA-treated cells were not significantly different to control cells. Increasing does of EPA were correlated with increasing levels of intracellular malondialdehyde (MDA). These observations suggest that EPA may influence the growth parameters of macrophages whilst inducing moderately elevated levels of oxidative stress. PMID- 10841045 TI - Generation of transgenic mice and germline transmission of a mammalian artificial chromosome introduced into embryos by pronuclear microinjection. AB - We have generated transgenic mice by pronuclear microinjection of a murine satellite DNA-based artificial chromosome (SATAC). As 50% of the founder progeny were SATAC-positive, this demonstrates that SATAC transmission through the germline had occurred. FISH analyses of metaphase chromosomes from mitogen activated peripheral blood lymphocytes from both the founder and progeny revealed that the SATAC was maintained as a discrete chromosome and that it had not integrated into an endogenous chromosome. To our knowledge, this is the first report of the germline transmission of a genetically engineered mammalian artificial chromosome within transgenic animals generated through pronuclear microinjection. We have also shown that murine SATACs can be similarly introduced into bovine embryos. The use of embryo microinjection to generate transgenic mammals carrying genetically engineered chromosomes provides a novel method by which the unique advantages of chromosome-based gene delivery systems can be exploited. PMID- 10841046 TI - Chromosomal location and nucleotide sequences of 5S ribosomal DNA of two cyprinid species (Osteichthyes, Pisces). AB - 5S ribosomal DNAs (rDNAs) from two cyprinid species, Acheilognathus tabira subsp. 1 and Cyprinus carpio, were isolated and sequenced. Tandemly arranged rDNAs were 179 bp in A. tabira and 204 bp in C. carpio. The non-transcribed spacer region elucidates the size difference of 5S rDNA between the two species. Fluorescence in-situ hybridization (FISH) localized 5S rDNAs to the short arms of two pairs of chromosomes in A. tabira and two to four pairs in C. carpio. Subsequent analysis demonstrated NORs in one pair of chromosomes in both species. Both the NOR and 5S rDNA are carried by a chromosome pair in A. tabira, but they are located on different chromosomes separately in C. carpio. Karyotype evolution by tetraploidy seems complex in cyprinid species. PMID- 10841047 TI - A novel interspersed type of organization of satellite DNAs in Tribolium madens heterochromatin. AB - Analysis of arrangement of satellite DNA sequences in Tribolium madens (Insecta, Coleoptera) by Southern analysis of pulsed-field blots and two colour FISH on extended chromosomes and DNA fibres revealed a novel type of heterochromatin organization. Two satellite DNAs, distributed over the whole pericentromeric heterochromatin of all chromosomes form clusters, ranging in size from 150 kb up to several Mb. Within the clusters, both satellites are in the form of highly interspersed, short homogeneous arrays which vary in size with a lowest length limit of only few kb. The longest arrays composed of a single satellite are relatively short, up to 70 kb for satellite I, and up to 45 kb for satellite II. Only a small fraction of about 15% of satellite II is organized in long tandem repeats, while the rest is in the form of only a few repeats intermingled with satellite I. The results indicate that large clusters composed of interspersed arrays of both satellites represent a major component of T. madens heterochromatin, which is mostly devoid of long regions of other sequences. The same organizational pattern probably also includes a region of the functional centromere. We propose that such an organizational pattern of DNA sequences in heterochromatin might be common in genomes characterized by a high rate of interchromosomal exchange. This pattern of organization is different from that in other animal as well as plant species analysed up to now, in which every satellite in heterochromatin is organized in a small number of large separate domains. PMID- 10841048 TI - Major and 5S ribosomal sequences of the largemouth bass Micropterus salmoides (Perciformes, Centrarchidae) are localized in GC-rich regions of the genome. AB - Major and 5S ribosomal genes have been localized in the chromosomes of Micropterus salmoides. By C-banding, Ag-staining, CMA3-staining and 45S and 5S fluorescence in-situ hybridization (FISH), we demonstrate that the 45S and 5S ribosomal genes are clustered in two different chromosome pairs and both are located in heterochromatic GC-rich regions. PCR amplification and sequencing of the 5S intergenic non-transcribed sequences have allowed us to identify variability essentially due to a trinucleotide tandem repeat (GCT). PMID- 10841049 TI - Comparative chromosome map of the laboratory mouse and Chinese hamster defined by reciprocal chromosome painting. AB - Cross-species reciprocal chromosome painting was used to determine homologous chromosomal regions between the laboratory mouse and Chinese hamster. When mouse chromosome-specific paints were hybridized to Chinese hamster chromosomes, paints specific for mouse chromosomes 3, 4, 9, 14, 18, 19 and X each painted a single chromosomal region, whilst other mouse paints delineated multiple discrete chromosomal regions. The mouse Y paint produced non-specific signals on Chinese hamster chromosomes. Nineteen mouse autosome paints identified a total of 47 homologous chromosome regions in the genome of the Chinese hamster. Hybridization of Chinese hamster paints to mouse chromosomes not only confirmed the above results, but also identified which of the chromosomal regions of these two species were homologous. In total, 10 Chinese hamster autosomal paints detected 38 homologous autosomal segments in the mouse genome. A comparative chromosome map was established based on these reciprocal chromosome painting patterns. This map forms the basis for exchanging gene mapping information between the species and for studying genome evolution. PMID- 10841050 TI - Differentiation and the polymorphic nature of the Y chromosomes revealed by repetitive sequences in the dioecious plant, Rumex acetosa. AB - The dioecious plant Rumex acetosa has a multiple sex chromosome system: females are 2n = XX + 12, males are 2n = XY1Y2 + 12, and the two Y chromosomes are heterochromatic. A DNA sequence abounded in the mare genome was isolated and analyzed. The sequence (RAE180) was a 180-bp-long tandemly arranged repetitive sequence, distributed in chromosomes Y1 and Y2, and two pairs of autosomes. Both Y chromosomes contained large amounts of RAE180 and the sequence formed many DAPI bands, while, on the two pairs of autosomes, RAE180 did not form DAPI bands. The internal structure and morphological changes of the Y chromosomes were analyzed by FISH, using RAE180 and the Y-chromosome-specific sequence RAYSI as probes. The pattern of the FISH signals caused by the accumulation of RAE180 and RAYSI suggested the structural change in the Y chromosomes during the process of sex chromosome evolution, and the morphological change in the Y chromosomes was explained by reciprocal translocation and inversion. PMID- 10841051 TI - Cytogenetic analysis of the pufferfish Tetraodon fluviatilis (Osteichthyes). AB - Because of their compact genome, pufferfish (Tetraodontiformes) have been proposed as a model for the study of the vertebrate genome. The genome of pufferfish is peculiar as it has the structural complexity of the genomes of higher vertebrates, but has small introns and lacks large clusters of highly repetitive sequences. Despite such interest, information about the genetics of pufferfish is still scanty. To fill this gap, we have performed a cytogenetic analysis of the pufferfish, Tetraodon fluviatilis, which can be maintained in an aquarium for a long time and, unlike the pufferfish, Fugu rubripes, it is not difficult to obtain. Karyotype analysis shows that T. fluviatilis has 2n = 42 with two metacentric chromosomes, four submetacentrics, two subtelocentrics and 34 acrocentrics. C-banding, followed by DAPI staining, showed that heterochromatin is essentially AT-rich and is located at centromeres. Staining of the same metaphase plates with CMA3 showed the presence of four heterochromatic regions located on two pairs of submetacentric chromosomes. Silver staining and FISH with a 28S rDNA probe showed that these GC-rich regions are nucleolar organizing regions (NORs). Finally, regardless of the technique used, no difference in the chromosome complement was found between males and females. PMID- 10841052 TI - Chromosome no. 1 of Crepis capillaris shows defined 3D-shapes in mitotic prophase. AB - The shape of mitotic prophase chromosomes has been studied in root tip nuclei by confocal microscopy and 3D-image analysis. Crepis capillaris chromosome no. 1 was used as a test object. Chromosome conformation was studied in early, mid- and in late prophase. In mid- and late prophase, individual chromosomes could be distinguished on the basis of their length. Early prophase chromosomes could not be distinguished as individuals. The central axes of prophase chromosomes were traced with an automated computer procedure and then represented as a string of 3D coordinates. This representation facilitated measurement along the chromosome axis of shape parameters such as curvature (amount of bending), torsion (helical winding) and torsion sign (helical handedness). Stretches of early prophase chromosomes showed full helical turns, which could be left- or right-handed. In the later prophase stages curvature and torsion were statistically analysed. Our data on 40 midprophase chromosomes no. 1 show that they are still highly curved, but full helical turns were no longer found. Instead, an overall meandering pattern was observed. In late prophase, one central loop persisted, flanked by two preferential regions of high curvature. PMID- 10841053 TI - A comparative chromosome map of the Arctic fox, red fox and dog defined by chromosome painting and high resolution G-banding. AB - A complete set of paint probes, with each probe specific for a single type of dog chromosome, was generated by DOP-PCR amplification of flow-sorted chromosomes. These probes have been assigned to high-resolution G-banded chromosomes of the dog and Arctic fox by fluorescence in-situ hybridization. On the basis of these results we propose improved nomenclature for the G-banded karyotypes of the dog and Artic fox. A comparative map between the Arctic fox, red fox and dog has been established based on results from chromosome painting and high-resolution G banding. This map demonstrates that the euchromatic complements of these three canid species consists of 42 conserved segments. Thirty-four of these 42 segments are each represented by a single dog chromosome with dog chromosomes 1, 13, 18 and 19 each retaining two segments, respectively. The autosomes of the Arctic fox and red fox could be reconstructed from these 42 blocks in different combinations through chromosomal fusions. Our findings suggest that chromosome fusion has been the principal mechanism of karyotype evolution occuring during speciation in canids. PMID- 10841054 TI - Cytogenetic studies of Hynobiidae (Urodela) XVI. Comparative C-banded karyotype analysis of Pseudohynobius flavomaculatus (Fei et Ye), Ranodon shihi (Liu) and Batrachuperus pinchonii (David). AB - Initial analysis of Pseudohynobius flavomaculatus chromosomes determined the chromosome number of this species to be 2n = 52. A re-examination of Ranodon shihi chromosomes detected 2n = 66 chromosomes, in contrast with a previous finding of 2n = 64. The C-banding patterns of these two species and that of Batrachuperus pinchonii were compared with each other. Regions of homoeology in the C-banding pattern among these three species represented 33.51-48.30% of the total length of their chromosomes. We also detected two types of chromosome rearrangement in hynobiid species based on the results of the present and previous cytogenetic studies. PMID- 10841055 TI - Maize nac1 and cld genes map to chromosome arms 10L and 2S, and to 4L and 5L, respectively. PMID- 10841056 TI - Power Doppler scanning in the diagnosis of carotid body tumors. AB - The aim of this work was to show contribution of power Doppler imaging in the diagnosis of the carotid body tumors. Six patients with a nontender mass beneath the mandibular angle were evaluated with gray scale and power Doppler sonography. Well-defined, solid, weakly hyperechoic masses were noted on gray scale sonography in the carotid bifurcation. Power Doppler sonography showed abundant flow, characterized as an intense blush, throughout the entire tumor in all patients. We believe that invasive and expensive diagnostic modalities are not necessary to evaluate carotid body tumors. Gray scale sonography and power Doppler imaging are sufficient for primary diagnosis of carotid body tumors. PMID- 10841057 TI - Sonographic features related to volvulus in neonatal intestinal malrotation. AB - This 3 year prospective study evaluated the sensitivity and specificity of abdominal ultrasonography and color Doppler ultrasonography in 31 neonates with suspected malrotation or malrotation with volvulus. Water instillation was used to detect duodenal dilatation, edema, and malrotated bowels. Twenty patients with ultrasonographic characteristics of inversion of the superior mesenteric artery and superior mesenteric vein were later surgically proved to have malrotation. Nine of these 20 patients also had volvulus. Sonographic features suggestive of volvulus included duodenal dilation with tapering configuration (8 of 9 cases, 89%), fixed midline bowel (8 of 9 cases, 89%), whirlpool sign (8 of 9 cases, 89%), and dilation of the distal superior mesenteric vein (5 of 5 cases, 100%). The sensitivity and specificity of duodenal dilation with tapering configuration for detecting volvulus were 89% and 92%, respectively; of fixed midline bowel, 89% and 92%; of whirlpool sign, 89% and 92%; and of dilation of distal superior mesenteric vein, 56% and 73%. The results of this study indicate that ultrasonographic features of inversion of the superior mesenteric artery and superior mesenteric vein could aid in the diagnosis of malrotation, and certain sonographic features can also be used to evaluate volvulus, a condition requiring emergent operation. PMID- 10841059 TI - Lipoma arborescens: high-resolution ultrasonographic findings. AB - Three patients with lipoma arborescens of the knee joint were evaluated with high resolution ultrasonography. All patients demonstrated a hyperechoic, frondlike mass in the suprapatellar bursa with a large associated effusion. All had popliteal cyst formation without involvement by the mass. Dynamic compression and manipulation of the suprapatellar effusion demonstrated pliable and flexible nature of mass with bending and waving of the frondlike mass in real time. Comparison with MR imaging demonstrated similar extent, location, and morphology of the mass. The characteristic features of this lesion as well as its distribution in the knee joint can be demonstrated readily by sonography. PMID- 10841058 TI - Sonographic differences in the appearance of acute and chronic full-thickness rotator cuff tears. AB - This study was undertaken to identify differences in the sonographic appearance of acute and chronic full-thickness rotator cuff tears. The ultrasonograms of 24 patients with an acute rotator cuff tear and 20 with a chronic tear were reviewed for tear size (width), location, and the presence and distribution of fluid. Among these 24 patients, 75% with a midsubstance tear location had an acute tear; 64% of patients with joint or bursal fluid had an acute tear; 80% of patients with a nonvisualized rotator cuff due to a massive tear had a chronic tear; and 73% of patients with no sonographic evidence of bursal or joint fluid had a chronic tear. In conclusion, a midsubstance location and the presence of joint or bursal fluid were more commonly associated with an acute tear. A nonvisualized cuff and the absence of joint and bursal fluid were more commonly observed with a chronic tear. PMID- 10841060 TI - Effect of spectacular reflection on out-of-plane ultrasonographic images reconstructed from three-dimensional data sets. AB - The effect of specular reflection on ultrasonographic images reconstructed out of plane to the plane of acquisition of a three-dimensional volumetric data set was studied using two in vitro phantoms that incorporated structures exhibiting specular reflection. The phantoms were scanned transversely (axially) to form three-dimensional data sets, with coronal cross-sectional images reconstructed perpendicular to the plane of acquisition of the data sets. Directly scanned, nonreconstructed coronal images of the phantoms also were obtained in the same planes and from the same areas as the reconstructed coronal images. The direct and reconstructed coronal images were compared. Owing to the inherent directionality of specular reflectors, the reconstructed coronal images differed from the directly scanned images in two ways, containing some hyperechoic regions that were not present at direct coronal scanning and failing to contain other hyperechoic areas that were present at direct coronal imaging. We conclude that sonographic images reformatted from volumetric data sets may have a different appearance than images scanned directly in the same plane, independent of other factors such as resolution. This should be taken into account when such reformatted images are interpreted. PMID- 10841061 TI - Comparability of the ultrasonic tissue characteristics of carotid plaques. AB - Hypoechoic carotid atherosclerotic plaques on ultrasonographic examination were found to be associated with cerebrovascular events. This underlines the need for an accurate evaluation of their echotexture characteristics. The objective of this study was to further validate a proposed method of normalization aiming to facilitate the comparability of these characteristics. The sonographic examination involved imaging of carotid plaques using duplex technique and capturing, digitization, and normalization in a computer in a standard way. In the first part, the interobserver and interscanner variability and the gain-level variability within the ultrasound unit were examined, before and after normalization, in terms of plaque echotexture. The second part investigated the effect of normalization on the echotexture of 419 symptomatic and asymptomatic plaques. Our results indicated that the normalization reduced the interscanner variability and the gain-level variability. The interobserver variability was excellent. In addition, this process further distinguished the echotexture characteristics of symptomatic and asymptomatic carotid plaques, which encourages the use of this method in natural history studies in persons with asymptomatic carotid atherosclerotic disease, aiming to identify those at higher risk for stroke. These patients might benefit from a carotid endarterectomy. PMID- 10841062 TI - Obstetrical sonography: the best way to terrify a pregnant woman. PMID- 10841063 TI - Ultrasonography improves diagnostic accuracy of acute appendicitis and provides cost savings to hospitals in Japan. AB - A cost analysis based on changes in patient care was used to evaluate the utility of abdominal ultrasonography in both the clinical management of patients clinically suspected of having acute appendicitis and in reducing expenditure of hospital resources. Among the 200 patients suspected of having acute appendicitis, 57 actually had acute appendicitis. Interpretation of appendiceal ultrasonographic results was 98.5% accurate. The ultrasonographic result led to changes in the treatment of 103 patients. Moreover, ultrasonography led to the prevention of unnecessary appendectomy in 25 patients, providing a savings to the hospital of about Yen 8,013,450 ($65,150), and prevented unnecessary hospital admission for 78 patient-days, thus saving the hospital approximately Yen 1,199,250 ($9750). The cost of performing the 200 ultrasonographic examinations was about Yen 1,096,176 ($8912), and thus the overall savings to the hospital was approximately Yen 40,590 ($330) per patient. Ultrasonography performed in patients with suspected acute appendicitis improves patient diagnostic accuracy, thus leading to more appropriate selection of patient treatment and reduced hospital expenditure. PMID- 10841064 TI - Sonohysterographic findings of endometrial malacoplakia. PMID- 10841065 TI - Prenatal sonographic diagnosis of Larsen syndrome. PMID- 10841066 TI - Cystic mesothelioma of the testis in an adolescent patient. PMID- 10841067 TI - [Rheumatic disorders. Overview]. AB - Of all rheumatic diseases, osteoarthritis (OA) and rheumatoid arthritis (RA) are the most frequently occurring. Although they differ in pathophysiology and the molecular mechanisms responsible for the destruction of cartilage (since RA is an inflammatory disease and OA is not), there are, however, a certain number of similarities and common pathways in the inflammatory processes of both diseases: mild inflammatory phenomena have been observed during OA, and both interleukin-1 and tumour necrosis factor-alpha seem to play key roles, as in RA. Although there is a dramatic difference between the 2 diseases in the intensity of inflammation, the inflammatory process is responsible for the synthesis of metalloproteinases and free oxygen radicals, and, subsequently, for progressive cartilage destruction. Both OA and RA engender important costs for the healthcare system. Direct costs result from practitioner visits, drug purchase and management, drug related adverse effects, management or hospital care; indirect costs are linked to progressive functional disability. Although RA leads to significant individual costs, OA is more problematical for the healthcare system, since its prevalence is far higher than that of RA. Thus, rheumatic diseases have become a major public health problem. Optimal therapeutic strategies need to be determined in order to define the most effective procedure for controlling disease symptoms such as pain, stopping or slowing down disease progression and, finally, keeping patients active. But it is of paramount importance that the gain in efficacy be associated with a gain in drug safety. PMID- 10841068 TI - [Nonsteroidal anti-inflammatory agents in the management of rheumatic disorders]. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) have become one of the most commonly used drug classes. However, the gastrointestinal adverse effects, including ulcers, perforation and bleeding, constitute a major problem, although the risk of their occurrence appears to vary according to the different classical NSAIDs used. These drugs inhibit the key enzyme in prostaglandin biosynthesis, cyclo-oxygenase or COX. COX exists in 2 isomeric forms, COX-1 and COX-2, which are inhibited to different extents by the classical NSAIDs. COX-1 leads, for example, to the production of prostaglandins that are cytoprotective for the gastric mucosa. COX-2 is inducible in a number of cells by pro-inflammatory stimuli. The selective COX concept predicts that inhibition of COX-1 may result in gastrointestinal adverse effects, whereas inhibition of COX-2 may underlie the anti-inflammatory actions of NSAIDs. These observations have led to the development of new drugs with high inhibitory potency for COX-2 and low potency for COX-1. These selective COX-2 inhibitors appear to be promising in animal models and endoscopic studies in humans. If large clinical and epidemiological studies confirm that (i) selective COX-2 inhibitors are as effective as classical NSAIDs, (ii) the frequency of severe gastrointestinal complications is considerably reduced with these new drugs, and (iii) no unexpected complications of selective COX-2 inhibition are detected, then selective COX-2 inhibitors will prove to be a significant advance in the treatment of patients with rheumatic diseases. PMID- 10841069 TI - [Gastrointestinal tolerance of nonsteroidal anti-inflammatory agents]. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) are effective antipyretic, analgesic and anti-inflammatory agents. One of the major concerns regarding the use of these compounds is the incidence of gastrointestinal (GI) adverse effects, ranging from dyspepsia to the serious and potentially life threatening complications of ulcers, haemorrhages, and perforations. Thus, the prevention and/or treatment of upper GI damage is estimated to increase the overall cost of NSAID therapy by at least 40%. The pathogenesis of NSAID-induced gastroduodenal mucosal injury appears to involve both topical and systemic mechanisms. The former is related to the acidic nature of most NSAIDs, which promotes the accumulation of ionised molecules (ion trapping) within the mucosal cells. Topical mucosal injury may also occur as a result of biliary excretion of active NSAID metabolites. The systemic effect has, however, the predominant role. It is mediated through cyclo-oxygenase (COX) inhibition and a subsequent decrease in gastroprotective prostaglandins. Fortunately, 2 forms of COX enzymes, designated COX-1 and COX-2, have been recognised. COX-1 appears to function as a house keeping enzyme, whereas COX-2 is primarily induced by inflammatory stimuli and mitogens in various cells, including macrophages and synovial cells. Accordingly, the inhibition of COX-2 would result in anti-inflammatory effects, whereas gastroduodenal ulceration is thought to be related to the inhibition of COX-1. Animal data have suggested that nabumetone has a low ulcerogenic potential in comparison with other available NSAIDs. This feature was further supported by controlled clinical trials as well as epidemiological studies. The relative GI safety of nabumetone may be attributed to its lack of direct and indirect topical effects because of its nonacidic nature and absence of enterohepatic recirculation. Furthermore, the active metabolite [6-methoxy-2-naphthylacetic acid (6-MNA)] may be gastro-sparing as a result of its property of COX-2 preferential inhibition. PMID- 10841070 TI - [Properties and features of nabumetone]. AB - Nabumetone is a nonsteroidal anti-inflammatory drug (NSAID) of the 2,6 disubstituted naphthylalkanone class. It is a prodrug metabolised to an active metabolite, 6-methoxy-2-naphthylacetic acid (6-MNA), which preferentially inhibits cyclo-oxygenase-2 (COX-2) and has both anti-inflammatory and analgesic properties. The efficacy of nabumetone is comparable to that of other NSAIDs currently marketed in France for the treatment of rheumatoid arthritis and osteoarthritis. Nabumetone is metabolised to 6-MNA by extensive first-pass metabolism. This active metabolite is subsequently conjugated and excreted in urine. 6-MNA does not undergo enterohepatic recirculation, which may have implications for comparative gastrointestinal toxicity. Substantial concentrations of 6-MNA are achieved and maintained in synovial fluid, which is close to the proposed site of action in chronic arthropathies. The long plasma half-life of 6-MNA (20 to 24 hours) and its persistence in synovial fluid facilitate a once-daily dosage regimen. Nabumetone compares favourably with other NSAIDs with respect to its renal and gastrointestinal adverse effect profile, although increased clinical vigilance of drug-related adverse events relating to these organ systems is still recommended. In addition, in vitro experiments have shown this drug to have a good cartilage tolerability profile. Whether nabumetone has clinical utility in cardiovascular disorders remains to be determined. The unique pharmacokinetic and pharmacodynamic properties of nabumetone make it a welcome addition to the growing arsenal of drugs used in the treatment of chronic arthropathies. There may be clinical advantages in terms of the relative tolerability profile of nabumetone, given that this drug has a low COX-2/COX-1 ratio compared with other nonselective NSAIDs. PMID- 10841071 TI - [Rationale for using nabumetone and clinical experience]. AB - Nabumetone's position as one of the most commonly used nonsteroidal anti inflammatory drugs (NSAIDs) in the world today is based upon over a decade of clinical experience. The popularity of this drug lies in both its unique pharmacokinetic profile and special safety features in pharmacodynamic terms. This nonacidic prodrug with an active 6-methoxy-2-naphthylacetic acid (6-MNA) metabolite has COX-2 preferential features and is also devoid of enterohepatic recirculation. It is felt that these characteristics have provided the basis for its unique long term tolerability, documented in various at-risk osteoarthritis and rheumatoid arthritis populations. The excellent tolerability of nabumetone and its 24-hour half-life, which provides the advantages of a once-daily dosage regimen, make it uniquely suitable for long term anti-inflammatory therapy in arthritis. The tolerability profile of nabumetone has also demonstrated clear cost-effectiveness advantages, as confirmed by comparative and epidemiological studies. Selective COX-2 NSAIDs are likely to prove more expensive because of the increasing costs and demands of clinical research prior to FDA approval. These higher costs may limit and influence patient access, depending on the healthcare delivery system, and many years of experience will be required to document the putative tolerability advantages of these newer COX-2 inhibitor agents. In the meantime, it is comforting that nabumetone has established such an advantageous tolerability profile together with acknowledged efficacy. PMID- 10841072 TI - Molecular mechanism of ultraviolet-induced keratinocyte apoptosis. AB - This article reviews advances in the study of the molecular mechanisms for ultraviolet (UV)-induced keratinocyte apoptosis, with particular reference to the cytokines tumor necrosis factor-alpha (TNF-alpha) and Fas ligand (FasL). TNF alpha and FasL induce their respective receptors and then activate caspase enzymes that are critically involved in the apoptotic process. This activation is further amplified by intracellular mitochondria-associated mechanisms. Using gene targeted knockout mice lacking either the TNF-Rp55 or the TNF-Rp75, we have shown that TNF-alpha plays an important role in UV-induced keratinocyte apoptosis via TNF-Rp55. TNF-Rp55 shares homology with Fas and contains an intracellular death domain. UV seems to directly stimulate cross-linking of Fas, resulting in the engagement of the death machinery. Fas-associated death domain protein (FADD) acts as an adapter protein in both the TNF-Rp55 and Fas death-inducing cascades and is responsible for downstream signal transduction by recruiting caspases. Moreover, signaling of p53 contributes to the induction of apoptosis by regulating Bcl-2 family expression and increasing surface Fas expression. In addition to induction mechanisms of apoptosis, there are numerous inhibitory molecules that play a role in restricting the apoptotic pathway. Thus, the ultimate determination of whether or not a cell undergoes apoptosis after UV radiation is based on the balance between agonist and antagonist pathways. PMID- 10841073 TI - The carboxyterminal domains of human IFN-alpha2 and IFN-alpha8 are antigenically homologous. AB - The antigenic properties of human hybrid IFN-alpha8(60)/alpha1(92)/alpha8 were compared with those of human IFN-alpha1 and IFN-alpha2 using monoclonal antibodies (mAb). Hybrid IFN demonstrated a significantly closer antigenic relationship to the subtype alpha2 than to the subtype alpha1. In particular, high homology was observed between antigenic structures located in the C-terminal domains (93-166) of IFN-alpha8 and IFN-alpha2, whereas the corresponding N terminal receptor-binding domains (30-53) showed distinct antigenic characteristics. The 100% homology between IFN-alpha8 and IFN-alpha2 in the region 114-131 (helix D) indicated the role of this region in formation of the common antigenic structure. In IFN-alpha8/1/8, this shared antigenic structure was important for antiviral activity and exhibited immunodominant properties, consistent with functional and antigenic properties of the corresponding structure in IFN-alpha2. Based on this antigenic homology, we suggest that IFN alpha8 and IFN-alpha2 are evolutionarily more closely related to each other than to IFN-alpha1. This study will contribute to a better understanding of evolutionary events in the human IFN-alpha family. PMID- 10841074 TI - Inhibitory effects of IFN-gamma and acyclovir on the glioblastoma cell cycle. AB - Glioblastoma multiforme is one of the most aggressive and frequently occurring forms of brain cancer. It originates from astrocytes and is characterized by a loss of cell cycle control frequently involving mutations in tumor suppressor genes, such as p53 and p16. Nucleoside analogs, such as acyclovir (ACV), are currently being used in the treatment of viral diseases, such as those caused by members of the herpes family. Further, ACV in combination with type I interferons (IFN) has been shown to be more effective at lower doses in treatment of viral diseases. We show here that ACV at high concentrations (up to 500 microg/ml) inhibited growth in tissue culture of the human glioblastoma cell lines T98G, SNB 19, and U-373 by as much as 68.3% while inhibiting normal human astrocytes by only 38.3%. Related to this, the tumor cells were more than sevenfold more efficient in phosphorylation of ACV to the active phosphate form than normal human astrocytes. Analogous to treatment of virus-infected cells, suboptimal concentrations of ACV were as effective as high concentrations when used in conjunction with low concentrations of IFN-gamma in inhibition of tumor cell growth. At the cellular level, ACV and IFN-gamma inhibited the cell cycle in both the G1 and S phases. The cooperative effect of ACV and IFN-gamma against the glioblastomas appears to be due to direct inhibition of DNA synthesis by ACV in the S phase of the cell cycle and induction by IFN-gamma of the tumor suppressor gene p21wAF1/CIP1, which in turn acts at the level of proliferating cell nuclear antigen (PCNA) and cyclin E/cyclin-dependent kinase 2 (Cdk2) binding and inhibition of function. These studies show that the combination of IFN-gamma and ACV at suboptimal concentrations elicits significant antiproliferative effects on the glioblastoma cell lines T98G, SNB-19, and U-373 while having very little effect on normal human astrocyte cell proliferation. PMID- 10841075 TI - Regulation of murine interleukin-10 production by dehydroepiandrosterone. AB - Dehydroepiandrosterone (DHEA), one of the predominant androgens secreted by the adrenal cortex, is a potential immunologic regulator. In this report, the effect of DHEA on interleukin-10 (IL-10) production was studied in vivo. Mice were injected s.c. with DHEA or DHEA-sulfate (DHEAS) ranging from 50 microg to 500 microg/g body weight. The serum was collected, and the spleen cells were isolated 48 h after treatment. Results indicate that treatment with DHEA or DHEAS significantly increases the serum level of IL-10. The spleen cells isolated from the DHEA-treated or DHEAS-treated mice also showed an increase in IL-10 secretion and mRNA expression after the cells were activated by concanavalin A (ConA). The maximal dose of DHEA for inducing IL-10 production was 250 microg/g body weight. As IL-10 is a potent differentiation factor of B lymphocytes, the possible role of DHEA in regulation of immunoglobulin (Ig) production was studied in vivo. Results indicated a significant increase in both serum level of Ig (IgG, IgM, IgA) and Ig secretion by spleen cells after the mice were treated with DHEA or DHEAS. Mice injected with both DHEA (250 microg/g body weight) and anti-IL-10 antibody (0.5 mg/g body weight) showed a significantly reduced DHEA-mediated increase in Ig production. Thus, DHEA might affect the function of B lymphocytes via stimulating IL-10 production. PMID- 10841076 TI - Structure-function study of the extracellular domain of the human type I interferon receptor (IFNAR)-1 subunit. AB - Despite accumulating information about the different effector molecules and signaling cascades that are invoked on interferon-alpha (IFN-alpha) binding to the type 1 IFN receptor, little is known about the specifics of the binding interactions between the ligand and the receptor complex. The IFN-alpha/beta receptor (IFNAR)-2 subunit of the IFN receptor is considered the primary binding chain of the receptor, yet it is clear that both receptor subunits, IFNAR-1 and IFNAR-2, cooperate in the high-affinity binding of IFN to the receptor complex. Earlier results from our laboratory suggested that an association of IFNAR-1 with membrane Galalpha1-4Gal-containing glycolipids facilitates receptor-mediated signaling. The data implicated amino acid residues in the SD100 domain of IFNAR-1 in the glycosphingolipid (GSL) modification of the type 1 IFN receptor. Interestingly, the human and murine counterparts of IFNAR-1 exhibit remarkable species specificity despite their considerable amino acid sequence identity. Certainly, those amino acid residues that effect GSL modification of IFNAR-1 are conserved between species, yet specific regions of IFNAR-1 that confer species specificity have not been defined. To delineate further the role of the IFNAR-1 SD100A domain in receptor function, a chimeric cDNA was assembled, in which the SD100A domain of the murine IFNAR-1 chain was replaced with the human sequence. This construct was expressed in IFNAR-1-/- mouse embryonic fibroblasts, and stable transfectants were established. Transfectants are fully sensitive to murine IFN-alpha4 treatment with respect to the induction of IFN-stimulated gene factor 3 (ISGF3) and sis-inducing factor (SIF) signal transducer and activator of transcription factor (Stat) complexes, exhibiting comparable levels of Stat activation to those observed in IFNAR-1-/- cells reconstituted with intact MuIFNAR-1. Similar results were obtained with IFN-induced antiviral and growth inhibitory responses. Viewed together, these data suggest that the SD100A domain of IFNAR-1 does not contribute to species-specific IFN binding. PMID- 10841077 TI - Chemokine gene adjuvants can modulate immune responses induced by DNA vaccines. AB - Nucleic acid immunization has been shown to induce both antigen-specific cellular and humoral immune responses in vivo. Moreover, immune responses induced by DNA immunization can be enhanced by the use of molecular adjuvants. For example, coadministration of costimulatory molecules (CD80 and CD86), proinflammatory cytokines (interleukin-1alpha [IL-1alpha], tumor necrosis factor-alpha [TNF alpha, and TNF-beta), Th1 cytokines (interleukin-2 [IL-2], IL-12, IL-15, and IL 18), Th2 cytokines (IL-4, IL-5, and IL-10), and granulocytes-macrophage colony stimulating factor (GM-CSF) with DNA vaccine constructs leads to modulation of the magnitude and direction (humoral or cellular) of the immune responses. To further engineer the immune response in vivo, we compared the induction and regulation of immune responses from the codelivery of chemokine (IL-8, interferon gamma-inducible protein-10 [gammaIP-10], macrophage inhibitory protein-1alpha [MIP-1alpha], and RANTES) genes with codelivery of cytokine genes. We found that as in cytokine gene codelivery, coimmunization with chemokine genes along with DNA immunogen constructs can modulate the direction and magnitude of induced immune responses. We observed that coimmunization with IL-8, gammaIP-10, and MIP 1alpha genes increased the antibody response. We also found that coinjection with IL-8, gammaIP-10, and RANTES resulted in a dramatic enhancement of T helper (Th) proliferation response. Furthermore, among all coinjection combinations, we found that RANTES coinjection caused a high level of cytotoxic lymphocyte (CTL) enhancement. This enhancement of CTL responses observed from the coinjection with RANTES was CD8+ T cell dependent. Together with earlier reports on the utility of coimmunizing immunologically important molecules with DNA immunogens, we demonstrate the potential of this strategy as an important tool for the development of more rationally designed vaccines. PMID- 10841078 TI - Interleukin-6 increases thrombopoietin production in human hepatoma cells HepG2 and Hep3B. AB - The concentration of circulating thrombopoietin (TPO) is relatively high in patients with thrombocytosis reactive to inflammatory diseases. We investigated whether immunomodulatory cytokines stimulate TPO synthesis in cultured human hepatoma cells (lines HepG2 and Hep3B), renal proximal tubular cells, and bone marrow fibroblasts. The effects of interleukins (IL) IL-1beta, IL-6, and IL-11 and of tumor necrosis factor-a (TNF-alpha) on the rate of TPO secretion were measured by ELISA. TPO mRNA levels were quantitated by competitive reverse transcription PCR. HepG2 and Hep3B cells produced significant amounts of TPO mRNA and TPO protein. Renal tubular cells synthesized less TPO, and in bone marrow fibroblasts, neither TPO mRNA nor TPO protein was detected. Only IL-6 affected TPO protein secretion, causing a 1.5-fold stimulation in HepG2 and Hep3B cells in 24-h incubation periods. The TPO mRNA content in these cells was doubled by IL-6 after 2, 6, or 24 h of stimulation. Thus, IL-6 could cause thrombocytosis in inflammatory disease partly by increasing hepatic TPO production. PMID- 10841079 TI - Tumor necrosis factor-alpha and TNF-beta inhibit clonogenicity of mobilized human hematopoietic progenitors. AB - We studied the effect of tumor necrosis factors (TNF-alpha and TNF-beta) on the clonogenicity of peripheral blood progenitor cells (PBPC) mobilized by granulocyte colony-stimulating factor (G-CSF). The cells were obtained by nine leukaphereses in patients with malignancies undergoing peripheral blood stem cell transplantation (PBSCT). Flow cytometry was performed to evaluate the number of CD34+ cells in peripheral blood and leukapheresis products. PBPC were grown in semisolid medium supplemented with human growth factors in the absence or presence of TNF at concentrations ranging from 0.1 to 10 ng/ml. Colonies were scored on day 14. TNF-alpha and TNF-beta suppressed colony formation in all cases studied. TNF-alpha inhibited the growth of colony-forming units-granulocyte macrophage (CFU-GM) at 1 and 10 ng/ml and burst-forming units-erythroid (BFU-E) at 10 ng/ml (p < 0.05), and TNF-beta inhibited the growth of CFU-GM and BFU-E at 1 and 10 ng/ml (p < 0.05). Thus, G-CSF-mobilized hematopoietic PBPC are highly sensitive to both TNF-alpha and TNF-beta. This finding should be taken in account when PBPC are handled ex vivo. PMID- 10841080 TI - VEGF/Flk-1 interaction, a requirement for malignant ascites recurrence. AB - Vascular endothelial growth factor (VEGF) plays an important role in the production of ascitic fluid associated with malignant tumor growth. In an experimental model for malignant ascites formation, mice were inoculated intraperitoneally with syngeneic mouse sarcoma tumor cells. Ascites development was not prevented by administering tumor necrosis factor (TNF) simultaneously with the tumor cell inoculation. When the malignant ascites was first drained and renewal of ascites was monitored, however, a TNF dose-dependent inhibition of ascitic fluid accumulation was observed. Northern blot analyses indicated transient downregulation by TNF on the expression of VEGF mRNA in tumor cells. Monoclonal antibody, (mAb) DC101 generated against the mouse VEGF receptor Flk-1 prevented the recurrence of malignant ascites in mice similar to TNF inhibition. In addition, exogenous soluble human Flt-1 used as an inhibitor of endogenous VEGF binding also inhibited ascites recurrence. These data demonstrate that the observed inhibitory effect of TNF on reestablishment of malignant ascites can be achieved equally by inhibition of the interaction of VEGF with its receptor Flk 1. PMID- 10841081 TI - Role of sarcolectin (SCL) and interferons in coordinated T cell clonal expansion. AB - T cell multiplication is attributed to the growth factor interleukin-2 (IL-2), which is, however, only activated when a specific cell membrane-bound receptor can be expressed. We found in all human sera tested a lectin that we described and called sarcolectin (SCL). SCL is a molecularly cloned 55-kDa protein that stimulates DNA synthesis in all immunocompetent cells and inhibits the interferon (IFN)-dependent antiviral state. SCL is excreted in conditioned medium of T cell cultures grown under serum-free conditions, where it can be demonstrated regularly by Western blots. In such cultures, in addition to SCL and IL-2, IFN gamma and IFN-alpha also can be found, likely as a feedback response to DNA stimulation. Considered together, the data suggest that coordinated clonal expansion of T cells is governed by SCL-IL-2, both which induce T cell proliferation and simultaneously activate IL-2 receptors. T cell replication is downregulated by the effect of feedback IFN-gamma and IFN-alpha. To initiate a new growth cycle, SCL is thought to block the residual IFN-dependent antiproliferative state. PMID- 10841082 TI - The long forced march: from hell to a life of science. PMID- 10841083 TI - Electron microscopy as a tool for identifying new pathogens. AB - Electron microscopy (EM) is still an important tool for the investigation of infectious diseases, despite the introduction of powerful new methods, mainly involving the polymerase chain reaction. Particularly in the field of parasitic protozoology associated with AIDS, where many new species of human pathogens have been recognized in tissue biopsies, EM remains an essential 'catch-all' diagnostic method. The resolved ultrastructural details of these newly recognized parasites allows a unique insight into the biology of these organisms. The information produced by EM is different, but complementary, to that provided by alternative methods. PMID- 10841084 TI - Molecular techniques for the detection and identification of new bacterial pathogens. PMID- 10841085 TI - Gram-negative bacterial protein secretion. PMID- 10841086 TI - The prevalence of hepatitis B and C in HIV-positive Greek patients: relationship to survival of deceased AIDS patients. AB - OBJECTIVES: To determine the prevalence of hepatitis viruses B (HBV) and C (HCV) co-infections in HIV-infected patients and the overall impact of these co infections on deceased AIDS patients survival. METHODS: One hundred and eighty one patients (159 males, 22 females) infected with HIV, attending an academic AIDS unit in Athens, Greece, constituted the study population. The study population consisted of 124 homo/bisexual men, 34 heterosexuals, 12 intravenous drug users (IDU) and 11 blood transfusion recipients. Virological markers tested for HBV infection included HBsAg, anti-HBs and total anti-HBc by enzyme-linked immunoassays. Detection of HCV antibodies was carried out by third generation enzyme-linked immunoassay, and repeatedly positive samples were further tested by a supplemental enzyme-linked immunoassay; only sera reactive by both methods were considered to be HCV-positive. RESULTS: The prevalence of HBV markers was 67.4%: 71.8% in homo/bisexuals, 35.3% in heterosexuals, 91.7% in IDUs and 90.9% in blood transfusion recipients (P = 0.00004). The prevalence of HCV antibodies was 13.8%: 8.1% in homo/bisexuals, 8.8% in heterosexuals, 58.3% in IDU and 45.5% in blood transfusion recipients (P<0.000001). The prevalence of HCV antibodies was not significantly higher in homo/bisexuals than in heterosexuals (P= 0.8). Coinfection with HBV or HCV, or both, did not influence the survival of deceased AIDS patients (n = 73). CONCLUSIONS: HBV infection was equally prevalent among homo/bisexuals and IDU with HIV infection, whereas HCV infection was more prevalent in IDU than in homo/bisexuals with HIV infection. The prevalence of HCV infection was equal among heterosexuals and homo/bisexuals, indicating that if sexual transmission of HCV occurs, homo/bisexuals are not at greater risk than heterosexuals. Finally, the survival of deceased AIDS patients was not affected by the presence of HBV and HCV co-infections. PMID- 10841087 TI - Clusters of new tuberculosis cases in North-west London: a survey from three hospitals based on IS6110 RFLP typing. AB - OBJECTIVES: The relative contributions of reactivation of latent infection and clusters of new infections to the overall incidence of tuberculosis in the U.K. is unknown. A study was carried out in North-West London to determine the feasibility of IS6110 RFLP strain typing as a tool to investigate the relative contributions of these two sources. METHODS: All available isolates of M. tuberculosis from specimens collected over a calendar year at three participating hospitals were typed by RFLP using an IS6110 probe. Isolates exhibiting a single band pattern were subject to further typing using an oligonucleotide direct repeat probe. Demographic and clinical information on cases was obtained from the National Survey of Tuberculosis Notifications in England and Wales and further information sought on clustered cases as identified by RFLP typing. RESULTS: Twenty-seven (23%) of the 118 cases had shared IS6110 RFLP patterns. Strains from nine cases had single band patterns, but these were all distinguishable from each other when subjected to further typing by direct repeat probe. The remaining 18 cases belonged to eight clusters. Epidemiological links were established between all the patients in each cluster. The likelihood of being in a cluster was increased in cases with pulmonary smear-positive disease. It was lower in cases of Indian Sub-continent ethnic origin. For 10 of the 18 clustered cases epidemiological links had not been established by conventional contact tracing. CONCLUSIONS: Investigation of the relative contributions of reactivation of latent infection and new infection is feasible in a UJK population, using IS6110 RFLP typing of M. tuberculosis isolates and epidemiological enquiries. This study in London identified clustered, presumably new cases, the majority of whom had not been linked epidemiologically. Comprehensive IS6110 RFLP typing of UK isolates would probably identify many clusters of incident tubercular infection. PMID- 10841088 TI - Detection of Mycoplasma fermentans in healthy students and patients with congenital immunodeficiency. AB - OBJECTIVES: To determine the prevalence of M. fermentans at different anatomical sites in healthy subjects and in patients with congenital immunodeficiency, and to determine whether haematogenous invasion occurs among the latter. METHODS: A polymerase chain reaction (PCR) assay was used to detect M.fermentans in throat swabs and urine specimens from healthy students, and from patients with congenital immunodeficiency. Peripheral blood mononuclear cells (PBMCs) from the latter group were also tested. RESULTS: Sixty-two students provided throat swabs, of which 11 (18%) were M. Jermentans-positive; 46 provided urine specimens, of which eight (17%) were positive. Of the 45 students who provided both throat and urine specimens, 12 (27%) had M. fermentans-positive samples; four in the throat and urine, four in the throat only and four in the urine only. Nineteen of the 20 patients with congenital immunodeficiency provided throat swabs, of which one (5%) was M. fermentans-positive; 19 also provided urine specimens, of which three (16%) were positive. All of the immunodeficient patients provided a PBMC sample, but none was positive. CONCLUSION: M. fermentans occurred frequently at mucosal sites in a healthy population and in subjects with congenital immunodeficiency. However, such a deficiency did not lead to overt haematogenous invasion. PMID- 10841089 TI - Carriage of penicillin-susceptible and non-susceptible pneumococci in healthy young children in Goteborg, Sweden. AB - OBJECTIVES: To study carriage of Streptococcus pneumoniae among healthy young children, determine the proportion of strains with decreased susceptibility to penicillin, and study possible risk factors for the carriage of penicillin resistant strains. METHODS: Between February 1996 and February 1997, 620 healthy, 18-month-old children in Goteborg, Sweden were screened for carriage of S. pneumoniae with decreased susceptibility to penicillin. Nasopharyngeal samples were obtained from children visiting child health centres for routine health control. RESULTS: Streptococus pneumoniae was found in 322 samples and 18 strains (5.6%, CI95 3.4; 8.8) of all pneumococci showed decreased susceptibility to penicillin G with minimum inhibitory concentrations (MICs) ranging from 0.125 to 1.0 mg/l. The proportion of strains with decreased susceptibility was similar to that found in a laboratory-based material (6%), from the same geographical area and time period. A majority of the children with strains with decreased susceptibility to penicillin (n = 11) were not attending day-care centres. CONCLUSIONS: The prevalence of S. pneumoniae with reduced susceptibility to penicillin is still low in unselected healthy Swedish children. PMID- 10841090 TI - Nosocomial enterococcal infections in children. AB - OBJECTIVES: To identify factors that are associated with an increased risk of nosocomial enterococcal infection in children. METHODS: A matched case-control study was conducted between January 1989 and July 1993 at the Children's National Medical Center, Washington DC. One control patient for each case was identified. Control patients did not have nosocomial enterococcal infections and were matched with cases on the basis of age and time of admission closest to the case within a three-month period. Data were collected from systematic review of patient medical records. One hundred and one study patients (cases) were matched with 101 control patients. A case was defined as a patient with enterococcal infection who met the Centers for Disease Control and Prevention criteria for nosocomial infection. Microbiology methods included isolation, identification, and antimicrobial susceptibility testing of enterococci from clinical specimens. RESULTS: Risk factors associated with nosocomial enterococcal infections were determined by multiple conditional logistic regression analyses of the cases and controls. Factors identified were placement of a central line, gastrointestinal tract pathology, and administration of multiple antimicrobial agents. The median duration of antimicrobial therapy prior to diagnosis of nosocomial enterococcal infection was approximately 1 week. CONCLUSION: The incidence of nosocomial enterococcal infections in children may be controlled by limiting the number of antimicrobial agents administered to hospitalized high risk patients. The importance of our findings is relevant in an era of increasing rates of antimicrobial resistance in nosocomial enterococcal infections. PMID- 10841091 TI - Tuberculosis in chronic renal failure in Jeddah. AB - OBJECTIVE: Immunocomprised patients with chronic renal failure haemodialysis have been reported to be at increased risk of developing tuberculosis. The purpose of this study was to monitor the development of tuberculosis in patients with chronic renal failure that have not yet been dialysed, presenting at the Renal Clinic of the King Khalid National Guard Hospital, Jeddah, Saudi Arabia. METHODS: Eighty patients with chronic renal failure were followed up for a period of 3 years and were carefully monitored for the development of tuberculosis. Mantoux test, chest radiograph and sputum examinations were performed at the beginning of the study and every 6 months thereafter. RESULTS: At the end of 3 years, eight out of 80 patients (10%) had developed tuberculosis, confirming the high incidence of tuberculosis in this group of patients. All the eight patients who developed tuberculosis before being started on dialysis had relatively low creatinine clearances. No particular underlying renal disease was associated with the development of tuberculosis. Four patients developed pulmonary tuberculosis, two renal tuberculosis and one each cervical tuberculous lymphadenitis and tuberculous meningitis. All patients responded satisfactorily to anti tuberculosis therapy, as diagnosis was established early. CONCLUSION: Routine screening for tuberculosis is recommended for patients with chronic renal failure presenting at Renal Units, and tuberculosis chemoprophylaxis should be considered for those undergoing haemodialysis, particularly in countries with a high incidence of tuberculosis. PMID- 10841092 TI - Specific subgroup B adenovirus diagnosis by PCR of the fibre gene. AB - OBJECTIVE: A highly sensitive and specific PCR assay targeting regions of the fibre gene was developed for the identification of subgroup B adenovirus strains. This is critical, since these adenovirus strains are frequently associated with severe respiratory infections in infants and new-borns. METHODS: Clinical samples from nasopharyngeal aspirates were analysed by PCR using several sets of primers corresponding to sequences of the gene coding for the fibre protein. RESULTS: The assay allowed the detection and identification of all the genotypes of adenovirus subgroup B, based on the size of the amplified product when analysed on polyacrilamide gel electrophoresis. Specifically, one set of primers was able to amplify DNA of subgroup B but not subgroup C and E viruses. CONCLUSION: The detection of adenovirus and the genotyping can be done on a routine basis by a PCR assay using the fibre gene as a target. The assay allows the identification of ADV subgroup B, including genotype 7h, which is the single most important viral pathogen associated with respiratory diseases in infants and young children in the southern part of South America. PMID- 10841093 TI - Causes of macrocytosis in HIV-infected patients not treated with zidovudine. Swiss HIV Cohort Study. AB - OBJECTIVE: Zidovudine is a well known cause of macrocytosis. However, many HIV infected patients develop macrocytosis even though they do not receive zidovudine. The aim of this case-control study was to evaluate other causes of high mean corpuscular volumes (MCV) in HIV infected patients. METHODS: Thirty patients with a MCV > or = 100 fl (cases) were compared to 60 randomly selected controls with MCV A< or = 99 fl, none of them receiving zidovudine. RESULTS: Sex ratio, age, mean CD4, prevalence of alcohol abuse and liver disease were similar in both groups. Vitamin B12 or folic acid levels were not decreased in patients with macrocytosis. In contrast, there was an association between macrocytosis and use of stavudine alone or in combination with another antiviral drug (28/30 cases, 15/60 controls, odds ratio 40.6 (95% confidence interval 5.1-325.2), P< 0.001). The haemoglobin concentration among cases (mean 13.5 g/dl) was similar to that of controls (mean 13.0 g/dl). CONCLUSIONS: Stavudine use increase the risk of macrocytosis more than 40-fold in HIV-infected patients who do not receive zidovudine. None of the stavudine-treated patients, however, developed anaemia or had to interrupt treatment because of this side effect. PMID- 10841094 TI - Antibody response to Burkholderia cepacia in patients with cystic fibrosis colonized with Burkholderia cepacia and Pseudomonas aeruginosa. AB - INTRODUCTION: This study was designed to determine the relationship between formation of serum antibodies to lipopolysaccharide (LPS) core antigen of Burkholderia cepacia and pulmonary colonization with B. cepacia and Pseudomonas aeruginosa in patients with cystic fibrosis (CF), and to define if an enhanced host humoral immune response to B. cepacia was related to a poor clinical outcome. METHODS: Serum IgG to B. cepacia LPS core antigen was measured in adult cystic fibrosis patients colonized with B. cepacia and P. aeruginosa, and serial titres were measured in 13 B. cepacia and 41 P. aeruginosa colonized patients followed prospectively over 18 months. RESULTS: The median B. cepacia antibody titre was significantly greater in the patients colonized with B. cepacia compared to those colonized with P. aeruginosa, a group which grew B. cepacia intermittently from their sputum. and nine healthy controls. The median antibody titre at recruitment into the study was significantly greater in patients who later went into exacerbations compared with those who remained clinically stable. but there was no difference between B. cepacia antibody titres in patients who died and those who survived the study duration. DISCUSSION: The degree of overlap of serum IgG levels to B. cepacia LPS core antigen in cystic fibrosis patients colonized with B. cepacia and P. aeruginosa does not allow this antibody to be used in a clinical context to define infection status. The magnitude of the humoral response to B. cepacia may influence occurrence of pulmonary exacerbations, but a more exuberant humoral immune response to B. cepacia core LPS is not the mechanism by which pulmonary deterioration occurs. PMID- 10841095 TI - Candidaemia in special care nurseries: comparison of albicans and parapsilosis infection. AB - OBJECTIVES: Candidaemia caused by Candida parapsilosis (CP) is being increasingly reported among infants in neonatal intensive care units (NICU). To assess relative severity, clinical manifestations of candidaemia caused by C. albicans (CA) and CP in a NICU were compared. METHODS: Between January 1994 and July 1997, episodes of candidaemia occurring among infants hospitalized in the NICU were identified in a children's hospital. The demographic characteristics, associated risk factors, clinical manifestations and outcome of the infants with CP fungaemia were collected and compared with those of the infants with CA fungaemia. RESULTS: Twenty-four episodes caused by CA and 22 episodes caused by CP were included in this study. No significant differences were found between the two groups for gestational age, birth weight, male gender, post-natal age at onset of candidaemia, frequency of antecedent neonatal events, prior duration of antibiotic therapy and hyperalimentation, as well as presence of central venous catheter (CVC). Infants with CA fungaemia were significantly more likely than those with CP fungaemia to present with hypoxaemia, bradycardia and respiratory distress requiring intubation, and have a longer prior duration of indwelling CVC and a higher dissemination rate. The eradication rate of candidaemia and overall case fatality rate were comparable in both groups. but CP fungaemia did not appear to cause acute lethal events. CONCLUSION: The presenting signs of CP fungaemia are relatively not so severe, but CP fungaemia, which is relatively difficult to eradicate, increases the morbidity and mortality of the infants. PMID- 10841096 TI - Prevalence of hepatitis C virus infection among injecting drug users in Glasgow 1990-1996: are current harm reduction strategies working? AB - OBJECTIVES: To determine the prevalence of HCV antibodies among injecting drug users and to gauge the effectiveness of needle/syringe exchange in preventing the transmission of HCV infection. METHODS: Between 1990-1994 and in 1996, annual cross-sectional surveys of injecting drug users in Glasgow were conducted. In order to ensure as representative a sample as possible, the 1949 respondents were recruited from both 'in-treatment' and 'out-of treatment' settings. Injectors were interviewed about their risk behaviours for blood-borne viruses and provided a saliva sample which was initially tested, anonymously, for HIV antibodies, and subsequently tested for hepatitis C infection. RESULTS: Among 1949 injectors, the prevalence of salivary antibodies, indicative of hepatitis C viraemia, was 61%(95%, confidence interval (CI) 59%-63%): the estimated prevalence of serum antibody positivity was 72%. Length of injecting, year of commencing drug injecting and the number of times in prison were predictive of antibody positivity. Thirty-one per cent of injectors who commenced their injecting after 1992, following the full establishment of needle/syringe exchange in the city, were salivary antibody positive, and the majority of their infections were acquired outside the prison setting. Respondents who began injecting after the introduction of needle/syringe exchange in the city were significantly less likely to test HCV antibody positive than those who commenced injecting prior to the advent of needle/syringe exchange, after adjusting for length of injecting career. CONCLUSION: The prevalence of HCV among injectors in Glasgow has decreased during the era of needle/syringe exchange. However, there is evidence to suggest that the incidence of infection remains high. Since the prevalence of hepatitis C viraemia among the city's injecting population is extremely high, ongoing transmission is inevitable unless more effective interventions are identified and implemented urgently. PMID- 10841097 TI - Efficacy of long-term antibiotic suppressive therapy in proven or suspected infected abdominal aortic grafts. AB - We have reviewed our experience of long-term antibiotic suppressive therapy in patients who underwent repair of an abdominal aortic aneurysm (AAA) and developed proven or strongly suspected infection of a graft. Five patients with abdominal aortic repair complicated by proven or suspected graft infections were treated with continuing antibiotic suppressive therapy based on microbiology culture reports. Two patients developed infection of an established graft, two patients had a graft inserted into an infected area and one patient was thought to be at high risk of developing infection of a recently placed graft. All patients had severe co-existent medical problems and were considered too ill to tolerate further definitive surgery. Response to therapy was monitored by absence of symptoms, fever, inflammatory markers and survival. All patients are alive with a median survival of 32 months, the longest having survived for 6 years. In selected patients with abdominal aortic graft infections, indefinite antibiotic suppressive therapy may be an acceptable alternative to further surgery. PMID- 10841098 TI - Superior vena cava syndrome during the treatment of pulmonary tuberculosis in an HIV-1 infected patient. AB - Tuberculosis is nowadays an uncommon cause of superior vena cava syndrome (SVCS). We report the case of an HIV-infected patient who presented with respiratory symptoms accompanied by cervical and mediastinal lymphadenopathy. Sputum examination showed acid-fast bacilli, and treatment was instituted with isoniazid, rifampicin, pyrazinamide and ethambutol. A few days later SVCS developed, presumably as a consequence of inflammatory lymphadenitis. With corticosteroids, all symptoms disappeared. To our knowledge, no cases of SVCS provoked by this kind of paradoxical reaction have been described previously. PMID- 10841099 TI - Opsoclonus-myoclonus syndrome in a child with neuroborreliosis. AB - Opsoclonus-myoclonus is a rare neurological syndrome affecting children and adults. In children it occurs as a parainfectious process or a paraneoplastic syndrome in association with neuroblastoma. Here we report it presenting as an unusual neurological manifestation of Lyme borreliosis. To our knowledge, this is the first report which describes recovery from this syndrome in a child. PMID- 10841100 TI - Candida albicans meningitis: clinical case. AB - Candida spp. meningitis is still a rare clinical situation, although it is becoming more frequent. Literature references to it and therapeutic options are scarce. We present a case of a young male, HIV-positive drug addict, with Candida albicans meningitis which was treated with oral fluconazole, having a good outcome. PMID- 10841101 TI - Parvovirus B19 causing leucopenia and neutropenia in a healthy adult. AB - A 27-year-old healthy female presented with a prolonged and intermittent febrile illness. She was found to have leucopenia, neutropenia and thrombocytopenia, but a normal haemoglobin. The patient recovered spontaneously and convalescent serology 1 month later was positive for specific parvovirus B19 IgM and IgG. Parvovirus B19 infection was confirmed by detection of viral DNA by dot blot hybridization in a specimen of blood taken during the acute illness. A review of the previously reported cases of parvovirus B19-induced leucopenia in normal adults is presented. Parvovirus B19 should be considered in the differential diagnosis of leucopenia and neutropenia in healthy adults. PMID- 10841102 TI - Polymicrobial meningitis revealing an anterior sacral meningocele in a 23-year old woman. AB - Polymicrobial meningitis has become increasingly rare during recent decades. Historically, it has mainly been reported as being associated with disorders of the ENT-sphere. The treatment of these infections being optimized, polymicrobial meningitis nowadays is essentially a complication of gastrointestinal or gynaecological disorders and trauma. We present a case of polymicrobial meningitis following puncture of a unrecognized pre-sacral meningocele in a patient with Currarino syndrome and review of the relevant literature. PMID- 10841103 TI - A pseudolymphomatous skin reaction secondary to flucloxacillin. AB - We report the first case of a pseudolymphomatous skin reaction precipitated by flucloxacillin. Skin histology was suggestive of a cutaneous lymphoma, and DNA analysis by single stranded conformational polymorphism (SSCP) demonstrated T cell receptor gamma gene monoclonality. Withdrawal of flucloxacillin led to immediate clinical improvement and gradual resolution of skin rash and lymph nodes. PMID- 10841104 TI - Post-kala-azar dermal leishmaniasis during highly active antiretroviral therapy in an AIDS patient infected with Leishmania infantum. AB - We report a case of post-kala-azar dermal leishmaniasis (PKDL) in a woman with AIDS which occurred 13 months after a diagnosis of visceral leishmaniasis concomitantly with immunological recovery induced by highly active retroviral therapy. Cytokine pattern at the time of visceral leishmaniasis and PKDL diagnosis was studied and pathogenic implications were discussed. PMID- 10841105 TI - Disseminated fungal infection following falciparum malaria. AB - Falciparum malaria can cause immune suppression sufficient to allow opportunistic infection during the recovery phase. A patient is described who died from a disseminated infection with Aspergillus flavus and Absidia corymbifera, unresponsive to treatment with amphotericin and voriconazole. PMID- 10841106 TI - Peritonitis due to Ochrobactrum anthropi in a patient undergoing continuous ambulatory peritoneal dialysis. PMID- 10841107 TI - Cardiac tamponade caused by Nocardia asteroides in an HIV-infected patient. PMID- 10841108 TI - Nested PCR is useful to the clinician in the diagnosis of Pneumocystis carinii pneumonia. PMID- 10841109 TI - A cognitive task analysis for dental hygiene. AB - To be an effective assessment tool, a simulation-based examination must be able to evoke and interpret observable evidence about targeted knowledge, strategies, and skills in a manner that is logical and defensible. Dental Interactive Simulations Corporation's first assessment effort is the development of a scoring algorithm for a simulation-based dental hygiene initial licensure examination. The first phase in developing a scoring system is the completion of a cognitive task analysis (CTA) of the dental hygiene domain. In the first step of the CTA, a specifications map was generated to provide a framework of the tasks and knowledge that are important to the practice of dental hygiene. Using this framework, broad classes of behaviors that would tend to distinguish along the dental hygiene expert-novice continuum were identified. Nine paper-based cases were then designed with the expectation that the solutions of expert, competent, and novice dental hygienists would differ. Interviews were conducted with thirty one dental hygiene students/practitioners to capture solutions to the paper-based cases. Transcripts of the interviews were analyzed to identify performance features that distinguish among the interviewees on the basis of their expertise. These features were more detailed and empirically grounded than the originating broad classes and better serve to ground the design of a scoring system. The resulting performance features were collapsed into nine major categories: 1) gathering and using information, 2) formulating problems and investigating hypotheses, 3) communication and language, 4) scripting behavior, 5) ethics, 6) patient assessment, 7) treatment planning, 8) treatment, and 9) evaluation. The results of the CTA provide critical information for defining the necessary elements of a simulation-based dental hygiene examination. PMID- 10841110 TI - Observing the behavior of senior dental students in relation to infection control practices. AB - To achieve adequate infection control in the dental practice, dental students should be fully aware and prepared to adopt necessary procedures. But most studies of students' infection control behaviors rely on self-reported data. This study verified, through direct observation, how senior dental students performed basic infection control procedures and assessed how close to reality the reported behavior is to the observed behavior. Eight students were observed in three clinics: pediatric dentistry, minor oral surgery, and primary care promotion. The students were not aware of who was being observed or why. Later all twenty students comprising that clinical subgroup answered a questionnaire. Seventy-two patient-student contacts were observed. Students considered basic infection control procedures as relevant. The reported intention of adoption of these procedures after graduation dropped in relation to their reported relevance. Although the observed behavior in the three clinics was considered satisfactory, when compared to the reported behavior, it was found that the intention was more positive than the behavior itself. Even though the students were conscious about the importance of cross-infection control in the dental practice, the reported intention of future use and mainly the observed behavior should be improved. PMID- 10841111 TI - Teaching nightguard bleaching and other tooth-whitening procedures in North American dental schools. AB - Tooth-whitening using carbamide peroxide delivered in a custom-fitted tray (nightguard bleaching) is a relatively new procedure, yet it is currently one of the most commonly used types of esthetic dental treatment in private practice. This study determined the extent that nightguard bleaching (NGB) has been included in dental school curricula. All sixty-five dental schools in North America were surveyed about curriculum content and treatment protocol for the use of nightguard and other bleaching procedures, generating an 82 percent response. The survey covered eighteen subject areas related to NGB ranging from clinical requirements and indications to products and recall intervals used. The most commonly taught tooth-whitening procedure was NGB, which was most often taught by operative and restorative faculty. Although no schools had clinical requirements for NGB, 92 percent taught it. The most common indications for NGB were esthetic shade change and pre-restorative lightening of teeth. Unrestored caries, defective restorations, and pre-existing sensitivity were common contraindications. Most schools do not use a specific NGB consent form, but most use written patient instructions. Most schools use at least two different NGB products, bleach for two to four weeks, and use reservoired and scalloped trays. An average of 25 percent of NGB patients were estimated to develop sensitivity, for which treatment recommendations include fluoride, desensitizing toothpaste, and reduced exposure time. Curriculum time and safety concerns were reasons for not teaching NGB (8 percent schools). Most schools indicated that the relative importance of NGB in the curriculum was increasing. PMID- 10841112 TI - Student adjustment problems in two dental schools in Thailand. AB - Adjustment problems and academic achievement among dental students at two universities, one located in Thailand's capital city (Mahidol University) and the other in southern Thailand (Prince of Songkla University), were studied. All first- to sixth-year students at each university completed the Mooney Problem Checklist Thai version, by Supapan, which covers eleven areas of adjustment problems. The most severe adjustment problems at both schools were related to adjustment to college work. Problems relating to health and physical development, social and recreational activities, and moral and religious adjustment of MU dental students were significantly higher than those of PSU dental students. The checklist showed that, in both universities, the most important adjustment problem was the need for advice from staff concerning clinical experience. Stepwise regression indicated that four variables--adjustment to college work, sex, secondary education system and university (MU or PSU)--were associated with the academic achievement of dental students. PMID- 10841113 TI - An on-site social work program in an urban academic dental center. PMID- 10841114 TI - Implementation of portfolio assessment in a competency-based dental hygiene program. PMID- 10841115 TI - Effect of Shipman case on family practice. PMID- 10841116 TI - Is it time to abandon cardiotocographic ECG analysis? PMID- 10841117 TI - Early diagnosis of invasive aspergillosis. PMID- 10841118 TI - Transplantation for alcoholic liver disease in an era of organ shortage. PMID- 10841119 TI - Role of alcohol septal ablation in treatment of obstructive hypertrophic cardiomyopathy. PMID- 10841120 TI - Sensitive glucose sensing in diabetes. PMID- 10841121 TI - EUROCARE-2: relevance for assessment of quality of cancer services? PMID- 10841122 TI - A novel method for prediction of long-term outcome of women with T1a, T1b, and 10 14 mm invasive breast cancers: a prospective study. AB - BACKGROUND: Women with small mammographically detected breast cancers generally have good long-term outcomes, but a few with T1a (1-5 mm) and T1b (6-10 mm) tumours will eventually die from breast cancer. We investigated whether women at high risk of breast-cancer death could be identified with mammographic criteria and differentiated from women with small cancers of the breast and good outcomes. METHODS: We prospectively applied mammographic classifications of tumour type to a consecutive series of 343 mammograms of invasive breast cancers of size 1-14 mm. Classifications were: stellate (spiculated) mass with no calcifications; circular or oval lesions with no calcifications; spiculated or circular lesions with non-casting-type calcifications; and casting-type calcifications. FINDINGS: 20-year survival for women with 1-14 mm invasive tumours with casting-type calcifications was 55%. 14% of 138 women with 1-9 mm tumours had casting-type calcifications on mammography, which accounted for 73% of all breast-cancer deaths (p<0.001). T1a, T1b, and 10-14 mm tumours with casting-type calcifications behaved as if they were larger lesions, since the rate of death was similar to that for women with advanced high-grade tumours. Most women who died were node negative. The long-term survival of women who had tumours of 1-9 mm with no casting-type calcifications was about 95%. INTERPRETATION: Mammographic classification seemed to reliably predict good and bad long-term outcomes for survival in tumours of 14 mm or smaller, and especially for those smaller than 10 mm. The implications for therapy are substantial. PMID- 10841123 TI - Large-scale test of hypothesised associations between the angiotensin-converting enzyme insertion/deletion polymorphism and myocardial infarction in about 5000 cases and 6000 controls. International Studies of Infarct Survival (ISIS) Collaborators. AB - BACKGROUND: The original report of a possible association between myocardial infarction and the insertion/deletion (I/D) polymorphism of the gene for the angiotensin-1-converting enzyme (ACE) indicated a risk ratio for myocardial infarction with the DD genotype of 1.34 (95% CI 1.05-1.70), and the association was claimed to be particularly strong in a retrospectively defined low-risk subgroup (3.2 [95% CI 1.7-5.9). Subsequent investigations reached varying conclusions, but all were small, and much larger studies were needed. METHODS: 4629 myocardial infarction cases and 5934 controls were compared. Cases were UK men aged 30-54 years and women aged 30-64 years recruited on presentation to hospital with confirmed myocardial infarction. Controls were aged 30-64 years with no history of cardiovascular disease, but were siblings or children of myocardial infarction survivors, or spouses of such relatives. All risk-ratio calculations allow for this relatedness of some of the controls. An updated meta analysis of previous studies was also conducted. FINDINGS: The ACE DD genotype was found in 1359 (29.4%) of the myocardial infarction cases and in 1637 (27.6%) of the controls (risk ratio 1.10 [95% CI 1.00-1.21]). The association between myocardial infarction and the DD genotype did not seem to be stronger in the subgroup defined as low risk by previously used criteria (234 [28%] of 836 cases and 911 [28%] of 3253 controls: risk ratio 1.04 [95% CI 0.87-1.24]), or in any other subgroup. Nor was the ACE I/D genotype predictive of subsequent survival. INTERPRETATION: This study involved many more cases than any previously reported study of this question, but did not confirm the existence of any substantial association. In an updated meta-analysis of these results with those of previously published studies, the risk ratio for myocardial infarction with the DD genotype seems to lie in the range 1.0 to about 1.1. Although an increase in risk of up to about 10-15% cannot be ruled out, substantially more extreme risks can be. Moreover, there are not especially strong associations in the subgroups previously selected for emphasis. These findings illustrate the need for some studies of candidate genes to involve much larger populations than is customary, without undue emphasis on retrospectively defined subgroups. PMID- 10841124 TI - Effect of El Nino and ambient temperature on hospital admissions for diarrhoeal diseases in Peruvian children. AB - INTRODUCTION: To investigate whether the El Nino phenomenon and ambient temperature had an effect on the epidemiology of childhood diarrhoea, we analysed data on daily number of admissions of children with diarrhoea to the Oral Rehydration Unit of the Instituto de Salud del Nino in Lima, Peru, between January, 1993, and November, 1998. METHODS: We obtained daily data on hospital admissions from the Oral Rehydration Unit, and meteorological data from the Peruvian Weather Service, and used time-series linear regression models to assess the effects of the 1997-98 El Nino event on admissions for diarrhoea. FINDINGS: 57,331 children under 10 years old were admitted to the unit during the study. During the 1997-98 El Nino episode, mean ambient temperature in Lima increased up to 5 degrees C above normal, and the number of daily admissions for diarrhoea increased to 200% of the previous rate. 6225 excess admissions were attributable to El Nino, and these cost US$277,000. During the period before the El Nino episode, admissions for diarrhoea increased by 8% per 1 degree C increase in mean ambient temperature. The effects of El Nino and ambient temperature on the number of admissions for diarrhoea were greatest during the winter months. INTERPRETATION: El Nino had an effect on hospital admissions greater than that explained by the regular seasonal variability in ambient temperature. The excess increase in ambient temperature was the main environmental variable affecting admissions. If our findings are reproducible in other regions, diarrhoeal diseases may increase by millions of cases worldwide with each degree of increase in ambient temperature above normal. PMID- 10841125 TI - Effect of breastfeeding on infant and child mortality due to infectious diseases in less developed countries: a pooled analysis. WHO Collaborative Study Team on the Role of Breastfeeding on the Prevention of Infant Mortality. AB - BACKGROUND: The debate on breastfeeding in areas of high HIV prevalence has led to the development of simulation models that attempt to assess the risks and benefits associated with breastfeeding. An essential element of these simulations is the extent to which breastfeeding protects against infant and child mortality; however, few studies are available on this topic. We did a pooled analysis of studies that assessed the effect of not breastfeeding on the risk of death due to infectious diseases. METHODS: Studies were identified through consultations with experts in international health, and from a MEDLINE search for 1980-98. Using meta-analytical techniques, we assessed the protective effect of breastfeeding according to the age and sex of the infant, the cause of death, and the educational status of the mother. FINDINGS: We identified eight studies, data from six of which were available (from Brazil, The Gambia, Ghana, Pakistan, the Philippines, and Senegal). These studies provided information on 1223 deaths of children under two years of age. In the African studies, virtually all babies were breastfed well into the second year of life, making it impossible to include them in the analyses of infant mortality. On the basis of the other three studies, protection provided by breastmilk declined steadily with age during infancy (pooled odds ratios: 5.8 [95% CI 3.4-9.8] for infants <2 months of age, 4.1 [2.7-6.4] for 2-3-month-olds, 2.6 [1.6-3.9] for 4-5-month-olds, 1.8 [1.2-2.8] for 6-8-month-olds, and 1.4 [0.8-2.6] for 9-11-month-olds). In the first 6 months of life, protection against diarrhoea was substantially greater (odds ratio 6.1 [4.1-9.0]) than against deaths due to acute respiratory infections (2.4 [1.6 3.5]). However, for infants aged 6-11 months, similar levels of protection were observed (1.9 [1.2-3.1] and 2.5 [1.4-4.6], respectively). For second-year deaths, the pooled odds ratios from five studies ranged between 1.6 and 2.1. Protection was highest when maternal education was low. INTERPRETATION: These results may help shape policy decisions about feeding choices in the face of the HIV epidemic. Of particular relevance is the need to account for declining levels of protection with age in infancy, the continued protection afforded during the second year of life, and the question of the safety of breastmilk substitutes in families of low socioeconomic status. PMID- 10841126 TI - Cardiotocography only versus cardiotocography plus PR-interval analysis in intrapartum surveillance: a randomised, multicentre trial. FECG Study Group. AB - BACKGROUND: There is a need to improve the sensitivity and specificity of fetal monitoring during labour. We compared the gold standard, cardiotocography, with cardiotocography plus time-interval analysis of the fetal electrocardiogram in fetal surveillance. The aim was to find out whether time-interval analysis decreased the need for operative intervention due to fetal distress. METHODS: We did a randomised, prospective trial in five hospitals in the UK, Hong Kong, the Netherlands, and Singapore. 1038 women undergoing high-risk labours were randomly assigned fetal monitoring by cardiotocography alone, or cardiotocography plus fetal electrocardiography (ECG). Outcomes measured were rates of operative intervention, and neonatal outcome. Analysis was by intention to treat. FINDINGS: 515 women were assigned management by cardiotocography, and 523 cardiotocography plus fetal ECG. There was a trend towards fewer operative interventions for presumed fetal distress in the time-interval analysis plus cardiotocography group (63 [13%] vs 78 [16%]), but this was not significant (relative risk 0.80 [95% CI 0.59-1.08], p=0.17). There was no significant difference between groups in the proportion of babies who had an umbilical arterial pH of 7.15 or less (51 [11%] vs 49 [11%]; 1.01 [0.7-1.47]), or in the frequency of unsuspected acidaemia (42 [9%] vs 35 [8%]; 1.17 [0.76-1.79]). INTERPRETATION: The addition of time-interval analysis of the fetal electrocardiogram during labour did not show a significant benefit in decreasing operative intervention. There was no significant difference in neonatal outcome. PMID- 10841127 TI - Dissociation in people who have near-death experiences: out of their bodies or out of their minds? AB - BACKGROUND: Some people who come close to death report having experiences in which they transcend the boundaries of the ego and the confines of time and space. Such near-death experiences (NDEs) share some features with the phenomenon of dissociation, in which a person's self identity becomes detached from bodily sensation. This study explored the frequency of dissociative symptoms in people who had come close to death. METHODS: 96 individuals who had had self-reported NDEs, and 38 individuals who had come close to death but who had not had NDEs completed a mailed questionnaire that included a measure of "depth" of near-death experience (the NDE scale) and a measure of dissociative symptoms (the Dissociative Experiences Scale). Median scores in the two groups were compared with Mann-Whitney U tests. The association between depth of NDE and dissociative symptoms was tested by Spearman's rank-order correlation between scores on the NDE scale and the dissociative experiences scale. FINDINGS: People who reported NDEs also reported significantly more dissociative symptoms than did the comparison group. Among those who reported NDEs, the depth of the experience was positively correlated with dissociative symptoms, although the level of symptoms was substantially lower than that of patients with pathological dissociative disorders. INTERPRETATION: The pattern of dissociative symptoms reported by people who have had NDEs is consistent with a non-pathological dissociative response to stress, and not with a psychiatric disorder. A greater understanding of the mechanism of dissociation may shed further light on near-death and other mystical or transcendental experiences. PMID- 10841128 TI - Earache and back pain. PMID- 10841129 TI - Implementation of a specific approval process for blood-components prescription. AB - We implemented a systematic computer-assisted validation process for transfusion prescriptions to improve transfusion safety. Assessment of this new approach indicates good adoption of validated transfusion guidelines and a reduction of exposure to blood products and overall costs. PMID- 10841131 TI - Risk of cryptogenic fibrosing alveolitis in metal workers. AB - We report increased proportional mortality from cryptogenic fibrosing alveolitis in the workforce of a major UK engineering company. Measures of metal exposure from unbiased historical occupational records showed that among employees who have worked with metal, the risk of death from or with cryptogenic fibrosing alveolitis increased in relation to the duration of metal-working. PMID- 10841130 TI - Reversible ageusia as an effect of clopidogrel treatment. AB - In two patients we observed a loss of taste during clopidogrel treatment which was reversible. PMID- 10841132 TI - Hirudin treatment in a breastfeeding woman. AB - We report on a breastfeeding woman with deep venous thrombosis treated with hirudin because of heparin-induced thrombocytopenia, in whom hirudin was not detectable in human breastmilk. PMID- 10841133 TI - Salivary endothelin concentrations in the assessment of chronic heart failure. AB - We have shown that salivary endothelin concentrations are raised in patients with chronic heart failure and indicate progression of disease severity through each New York Heart Association Functional Class. Furthermore, endothelin concentrations in saliva seem to discriminate between controls and patients with mild symptoms of chronic heart failure. PMID- 10841134 TI - Exercise during growth and bone mineral density and fractures in old age. AB - If exercise is to be recommended during growth, benefits in bone mineral density (BMD) must be maintained in old age and shown to prevent fractures. Our cross sectional study of soccer players suggests that a high BMD is no longer recorded after retirement and fracture frequency is no less than predicted in old age. PMID- 10841135 TI - UK general practitioner guilty of killing 15 patients. PMID- 10841137 TI - Molecular assay helps predict lymphoma outcome PMID- 10841136 TI - New treatments proposed for chronic obstructive pulmonary disease. PMID- 10841138 TI - Magdi Yacoub: a surgeon and a scientist. PMID- 10841139 TI - Clinton's swan-song appeal for health-care reform. PMID- 10841140 TI - Controversy leads to new Netherlands HIV-test guidelines. PMID- 10841141 TI - Public and private bodies unite to push for global immunisation. PMID- 10841142 TI - Business response to HIV in South Africa. PMID- 10841143 TI - Lung cancer. AB - Lung cancer remains a major worldwide health problem, accounting for more than a sixth of cancer deaths. The proportion of cancers that are adenocarcinomas is increasing in North America and to some degree in Europe, leading to a changing clinical picture characterised by early development of metastases. Newer diagnostic techniques have allowed for more accurate tumour staging and treatment planning. In patients with non-small-cell cancer, surgical resection offers substantial cure rates in early-stage cases. Combined chemotherapy plus radiation therapy has clearly improved the treatment results for patients with locally advanced cancers, and patients with metastatic disease are now candidates for newer chemotherapy regimens with more favourable results than in the past. Small cell lung cancer is highly responsive to chemotherapy, and recent advances in radiation therapy have improved the prospects for long survival. New techniques for screening, and innovative approaches to both local and systemic treatment offer hope for substantial progress against this disease in the near future. PMID- 10841144 TI - Relation of nocturnal polyuria of the elderly to essential hypertension. AB - Nocturnal polyuria is common in the elderly. In this condition the normal circadian rhythm of urine production is reversed so that urine flow is higher at night than during the day. Elderly men with nocturnal polyuria are commonly referred for prostate surgery, which, not surprisingly, fails to relieve their symptoms. Compared with controls, patients with nocturnal polyuria have higher nocturnal sodium excretion but not higher nocturnal free-water clearance. Similar results have been obtained in children with nocturnal enuresis. Use of vasopressin analogues to induce water retention in elderly patients with nocturnal polyuria is illogical and potentially hazardous; nocturia can be more safely alleviated by diuretic therapy. Nocturnal polyuria in the elderly is associated with hypertension: this is consistent with studies in younger age groups that show that essential hypertension is associated with nocturia and with increased night/day ratios for sodium excretion. We propose that nocturnal polyuria and essential hypertension share some of the same pathophysiological determinants. Specifically, we suggest that a defect in the nitric-oxide pathway may lead to resetting of the pressure-natriuresis relation in the kidney, sodium retention, and compensatory nocturnal natriuresis. This suggestion is consistent with evidence that ageing and essential hypertension are both associated with defects in the nitric-oxide pathway. Our hypothesis has obvious therapeutic implications. More generally, studying the pathogenesis of nocturnal polyuria in the elderly may advance our understanding of the pathogenesis of essential hypertension. PMID- 10841145 TI - Evidence-based illiteracy: time to rescue "the literature". PMID- 10841146 TI - Stillbirths among offspring of male radiation workers. PMID- 10841147 TI - Stillbirths among offspring of male radiation workers. PMID- 10841148 TI - Stillbirths among offspring of male radiation workers. PMID- 10841149 TI - Thalamic perfusion in reflex sympathetic dystrophy syndrome. PMID- 10841150 TI - Thalamic perfusion in reflex sympathetic dystrophy syndrome. PMID- 10841151 TI - Thalamic perfusion in reflex sympathetic dystrophy syndrome. PMID- 10841152 TI - Prevalence of multiple system atrophy. PMID- 10841153 TI - Caesarean section and antiretroviral treatment. Italian trial on mode of delivery in HIV-positive women study group. PMID- 10841154 TI - Pathological and therapeutic distinctions in HUS/TTP. PMID- 10841155 TI - Pathological and therapeutic distinctions in HUS/TTP. PMID- 10841156 TI - Where is the mother in maternal and child health? PMID- 10841157 TI - Where is the mother in maternal and child health? PMID- 10841158 TI - Treatment of lympho-proliferative disease with rituximab. PMID- 10841159 TI - Outcome of renal transplantation. PMID- 10841160 TI - Cardiac risk stratification. PMID- 10841161 TI - The future of oncology. PMID- 10841162 TI - Is clinical examination an art? PMID- 10841163 TI - Doctors' attitudes to complementary medicine. PMID- 10841164 TI - Hypertension, migraine, and central retinopathy. PMID- 10841165 TI - Ergonomic microscope workstation. PMID- 10841166 TI - The Nobel chronicles. 1986: Stanley Cohen (b 1922); Rita Levi-Montalcini (b 1909). PMID- 10841167 TI - Use of oral corticosteroids and risk of fractures. AB - Treatment with oral corticosteroids is known to decrease bone density but there are few data on the attendant risk of fracture and on the reversibility of this risk after cessation of therapy. A retrospective cohort study was conducted in a general medical practice setting in the United Kingdom (using data from the General Practice Research Database [GPRD]). For each oral corticosteroid user aged 18 years or older, a control patient was selected randomly, who was matched by age, sex, and medical practice. The study comprised 244,235 oral corticosteroid users and 244,235 controls. The average age was 57.1 years in the oral corticosteroid cohort and 56.9 years in the control cohort. In both cohorts 58.6% were female. The most frequent indication for treatment was respiratory disease (40%). The relative rate of nonvertebral fracture during oral corticosteroid treatment was 1.33 (95% confidence interval [CI], 1.29-1.38), that of hip fracture 1.61 (1.47-1.76), that of forearm fracture 1.09 (1.01-1.17), and that of vertebral fracture 2.60 (2.31-2.92). A dose dependence of fracture risk was observed. With a standardized daily dose of less than 2.5 mg prednisolone, hip fracture risk was 0.99 (0.82-1.20) relative to control, rising to 1.77 (1.55 2.02) at daily doses of 2.5-7.5 mg, and 2.27 (1.94-2.66) at doses of 7.5 mg or greater. For vertebral fracture, the relative rates were 1.55 (1.20-2.01), 2.59 (2.16-3.10), and 5.18 (4.25-6.31), respectively. All fracture risks declined toward baseline rapidly after cessation of oral corticosteroid treatment. These results quantify the increased fracture risk during oral corticosteroid therapy, with greater effects on the hip and spine than forearm. They also suggest a rapid offset of this increased fracture risk on cessation of therapy, which has implications for the use of preventative agents against bone loss in patients at highest risk. PMID- 10841168 TI - Corticosteroids and fractures: a close encounter of the third cell kind. PMID- 10841169 TI - Efficacy and safety of daily risedronate in the treatment of corticosteroid induced osteoporosis in men and women: a randomized trial. European Corticosteroid-Induced Osteoporosis Treatment Study. AB - Long-term use of high-dose corticosteroids often results in bone loss, which may lead to osteoporosis-related fractures. This was a multicenter, double-blind study in which 290 ambulatory men and women receiving high-dose oral corticosteroid therapy (prednisone > or = 7.5 mg/day or equivalent) for 6 or more months were randomized to receive placebo, risedronate 2.5 mg/day, or risedronate 5 mg/day for 12 months. All patients received calcium 1 g and vitamin D 400 IU daily. The primary endpoint was lumbar spine bone mineral density (BMD) at month 12. Additional measurements included BMD at the femoral neck and trochanter and the incidence of vertebral fractures. Overall, there were statistically significant treatment effects on BMD at 12 months at the lumbar spine (p < 0.001), femoral neck (p = 0.004), and trochanter (p = 0.010). Risedronate 5 mg increased BMD at 12 months by a mean (SEM) of 2.9% (0.49%) at the lumbar spine, 1.8% (0.46%) at the femoral neck, and 2.4% (0.54%) at the trochanter, whereas BMD was maintained only in the control group. Although not powered to show fracture efficacy, we observed a reduction in the incidence of vertebral fractures of 70% in the combined risedronate treatment groups, relative to placebo (p = 0.042). Risedronate was well tolerated, had a good safety profile, and was not associated with gastrointestinal adverse events. We conclude that risedronate increases BMD and potentially reduces the incidence of vertebral fractures in patients with corticosteroid-induced osteoporosis. PMID- 10841170 TI - Temporal expression of the chondrogenic and angiogenic growth factor CYR61 during fracture repair. AB - The repair of a fractured bone is a complex biological event that essentially recapitulates embryonic development and requires the activity of a number of different cell types undergoing proliferation, migration, adhesion, and differentiation, while at the same time expressing a host of different genes. To identify such genes, we employed differential display and compared messenger RNA (mRNA) populations isolated from postfracture (PF) day 5 calluses to those of intact rat femurs. One such gene in which expression was up-regulated at PF day 5 is identified as CYR61, a member of the CCN family of secreted regulatory proteins. CYR61 is a growth factor that stimulates chondrogenesis and angiogenesis. We show that its mRNA expression during fracture repair is regulated temporally, with elevated levels seen as early as PF day 3 and day 5, rising dramatically at PF day 7 and day 10, and finally declining at PF day 14 and day 21. At the highest peak of expression (PF day 7 and day 10, which correlates with chondrogenesis), CYR61 mRNA levels are approximately 10-fold higher than those detected in intact femurs. Similarly, high protein levels are detected throughout the reparative phase of the callus, particularly in fibrous tissue and periosteum, and in proliferating chondrocytes, osteoblasts, and immature osteocytes. The secreted form of CYR61 also was detected within the newly made osteoid. No labeling was detected in hypertrophic chondrocytes or in mature cortical osteocytes. These results suggest that CYR61 plays a significant role in cartilage and bone formation and may serve as an important regulator of fracture healing. PMID- 10841171 TI - Human osteoblastic cells propagate intercellular calcium signals by two different mechanisms. AB - Effective bone remodeling requires the coordination of bone matrix deposition by osteoblastic cells, which may occur via soluble mediators or via direct intercellular communication. We have previously identified two mechanisms by which rat osteoblastic cell lines coordinate calcium signaling among cells: autocrine activation of P2 (purinergic) receptors leading to release of intracellular calcium stores, and gap junction-mediated communication resulting in influx of extracellular calcium. In the current work we asked whether human osteoblastic cells (HOB) were capable of mechanically induced intercellular calcium signaling, and if so, by which mechanisms. Upon mechanical stimulation, human osteoblasts propagated fast intercellular calcium waves, which required activation of P2 receptors and release of intracellular calcium stores but did not require calcium influx or gap junctional communication. After the fast intercellular calcium waves were blocked, we observed slower calcium waves that were dependent on gap junctional communication and influx of extracellular calcium. These results show that human osteoblastic cells can propagate calcium signals from cell to cell by two markedly different mechanisms and suggest that these two pathways may serve different purposes in coordinating osteoblast functions. PMID- 10841172 TI - Nuclear localization of the type 1 PTH/PTHrP receptor in rat tissues. AB - The localization of PTH/PTH-related peptide (PTHrP) receptor (PTHR) has traditionally been performed by autoradiography. Specific polyclonal antibodies to peptides unique to the PTHR are now available, which allow a more precise localization of the receptor in cells and tissues. We optimized the IHC procedure for the rat PTHR using 5-microm sections of paraffin-embedded rat kidney, liver, small intestine, uterus, and ovary. Adjacent sections were analyzed for the presence of PTHR mRNA (by in situ hybridization) and PTHrP peptide. A typical pattern of staining for both receptor protein and mRNA was observed in kidney in cells lining the proximal tubules and collecting ducts. In uterus and gut, the receptor and its mRNA are present in smooth muscle layers (PTHrP target) and in glandular cuboidal cells and surface columnar epithelium. This suggests that PTH, or more likely PTHrP, plays a role in surface/secretory epithelia that is as yet undefined. In the ovary, PTHR was readily detectable in the thecal layer of large antral follicles and oocytes, and was present in the cytoplasm and/or nucleus of granulosa cells, regions that also contained receptor transcripts. PTHR protein and mRNA were found in the liver in large hepatocytes radiating outward from central veins. Immunoreactive cells were also present around the periphery of the liver but not within two or three cell layers of the surface. Clear nuclear localization of the receptor protein was present in liver cells in addition to the expected cytoplasmic/peripheral staining. PTHR immunoreactivity was present in the nucleus of some cells in every tissue examined. RT-PCR confirmed the presence of PTHR transcripts in these same tissues. Examination of the hindlimbs of PTHR gene-ablated mice showed no reaction to this antibody, whereas hindlimbs from their wild-type littermates stained positively. The results emphasize that the PTHR is highly expressed in diverse tissues and, in addition, show that the receptor protein itself can be localized to the cell nucleus. Nuclear localization of the receptor suggests that there is a role for PTH and/or PTHrP in the regulation of nuclear events, either on the physical environment (nucleoskeleton) or directly on gene expression. PMID- 10841173 TI - Expression of Indian hedgehog, parathyroid hormone-related protein, and their receptors in the postnatal growth plate of the rat: evidence for a locally acting growth restraining feedback loop after birth. AB - A locally acting growth restraining feedback loop has been identified in the murine embryonic growth plate in which the level of parathyroid hormone-related peptide (PTHrP) expression regulates the pace of chondrocyte differentiation. To date, it is largely unknown whether this feedback loop also regulates the pace of chondrocyte differentiation in the growth plate after birth. We therefore characterized the spatio-temporal expression of Indian hedgehog (IHH), PTHrP, and their receptors in the postnatal growth plate from female and male rats of 1, 4, 7, and 12 weeks of age. These stages are representative for early life and puberty in rats. Using semiquantitative reverse-transcription polymerase chain reaction (RT-PCR) on growth plate tissue, IHH and components of its receptor complex, patched (PTC) and smoothened (SMO), PTHrP and the type I PTH/PTHrP receptor messenger RNA (mRNA) were shown at all ages studied irrespective of gender. Using in situ hybridization, IHH, PTHrP, and PTH/PTHrP receptor mRNA were detected in prehypertrophic and hypertrophic chondrocytes in both sexes during development. In addition, especially in the younger age groups, faint expression of PTH/PTHrP receptor mRNA also was shown in stem cells and proliferative chondrocytes. Immunohistochemistry confirmed the observations made with in situ hybridization, by showing the presence of IHH, PTC, PTHrP, and PTH/PTHrP receptor protein in prehypertrophic and hypertrophic chondrocytes. In addition, staining for hedgehog, PTC, and PTHrP also was observed in growth plate stem cells. No differences in staining patterns were observed between the sexes. Furthermore, no mRNA or protein expression of the mentioned factors was detected in the perichondrium. Our data suggest that in contrast to the proposed feedback loop in the early embryonic growth plate, which requires the presence of the perichondrium, a feedback loop in the postnatal growth plate can be confined to the growth plate itself. In fact, two loops might exist: (1) a loop confined to the transition zone and early hypertrophic chondrocytes, which might in part be autocrine and (2) a loop involving the growth plate stem cells. PMID- 10841174 TI - Heterotopic ossification of degenerating rat skeletal muscle induced by adenovirus-mediated transfer of bone morphogenetic protein-2 gene. AB - In vivo gene transfer is a recently developed device for efficient delivery of a therapeutic recombinant protein. We formulated the hypothesis that a high level of expression of bone morphogenetic protein 2 (BMP-2) could be a future therapeutic modality in terms of inducing substantial bone formation in vivo. First, to test this hypothesis, adenoviruses carrying BMP-2 gene were directly injected into the soleus muscle of adult rat. The BMP-2 gene was successfully overexpressed in the target muscle by adenovirus-mediated transfer, whereas bone formation in and around the muscle failed to occur in this case. Second, to recruit putative osteoprogenitor cells, we then induced ischemic degeneration of the target muscle by orthotopically grafting it simultaneously with the gene transfer. The combination of BMP-2 gene transfer and orthotopic muscle grafting resulted in successful ossification of almost the whole grafted muscle, whereas neither muscle grafting alone nor the combination of muscle grafting and adenovirus-mediated transfer of reporter gene LacZ induced any bone formation in the muscle. The ossification process was evident by positive von Kossa staining of the histological sections and roentgenographical radio-opacity of the region. It was also found that the BMP-2 transgene overexpressed in grafted muscles inhibited muscle regeneration, which should otherwise follow the muscle degeneration. We further demonstrated an up-regulation of BMP receptor type IA in grafted muscles, suggesting its involvement in the bone-formation process. In conclusion, overexpression of BMP-2 gene induced massive heterotopic ossification in skeletal muscles under graft-induced ischemic degeneration, which possibly up regulates osteoprogenitor cells in situ. PMID- 10841175 TI - The ribosomal protein QM is expressed differentially during vertebrate endochondral bone development. AB - Endochondral ossification is a carefully coordinated developmental process that converts the cartilaginous model of the embryonic skeleton to bone with accompanying long bone growth. To identify genes that regulate this process we performed a complementary DNA (cDNA) subtractive hybridization of fetal bovine proliferative chondrocyte cDNA from epiphyseal cartilage cDNA. The subtracted product was used to screen a fetal bovine cartilage cDNA library. Ten percent of the clones identified encoded the bovine orthologue of the human ribosomal protein "QM." Northern and western blot analysis confirmed that QM was highly expressed by cells isolated from epiphyseal cartilage as opposed to proliferative chondrocytes. In contrast, no detectable difference in the expression of mRNA for the ribosomal protein S11 was detected. Immunohistochemical analysis of fetal bovine limb sections revealed that QM was not expressed by the majority of the epiphyseal chondrocytes but only by chondrocytes in close proximity to capillaries that had invaded the epiphyseal cartilage. Strongest QM expression was seen in osteoblasts in the diaphyseal region of the bone adjoining the growth plate, within the periosteum covering the growth plate and within secondary centers of ossification. Hypertrophic chondrocytes within the growth plate adjoining the periosteum also were positive for QM as were chondrocytes in the perichondrium adjoining the periosteum. In vitro investigation of the expression of QM revealed higher QM expression in nonmineralizing osteoblast and pericyte cultures as compared with mineralizing cultures. The in vivo and in vitro expression pattern of QM suggests that this protein may have a role in cell differentiation before mineralization. PMID- 10841176 TI - The association between heel ultrasound and hormone replacement therapy is modulated by a two-locus vitamin D and estrogen receptor genotype. AB - Evidence supports the role of estrogen deprivation in the process of bone remodeling and increased risk of fracture in postmenopausal women but little is known about the genetic basis of individual differences in response to therapy. In a cross-sectional study, 425 ambulatory postmenopausal French-Canadian women from Quebec (age range, 42-85 years old) were genotyped for a common Bsm I polymorphism at the vitamin D receptor (VDR) gene as well as a Pvu II polymorphism in the estrogen receptor (ESR1) gene. Heel ultrasound was determined by right calcaneal quantitative ultrasound (QUS) and results were expressed as an age-and-weight-adjusted stiffness index (heel SI z score). Our aim was to investigate the interaction between hormone-replacement therapy (HRT) and receptor genotypes in an effect on heel SI. Notably, a two-locus genotype (VDR bb/ESR-PP) present in 9.5% of women was responsible for over 30% of the total HRT related heel SI difference in the whole sample. Women bearing this combined VDR/ESR1 genotype who received HRT for more than 5 years had a 21% (1.25 SD) greater heel SI (p = 0.002) than those bearing the same genotype but who received HRT for <5 years. This may translate into a 2- to 3-fold difference in the risk of fracture. Although follow-up studies are needed, our findings suggest that QUS of the heel in postmenopausal women taking HRT is affected by variation in VDR and ESR1 loci, jointly. PMID- 10841177 TI - Androgen deficiency induces high turnover osteopenia in aged male rats: a sequential histomorphometric study. AB - Hypogonadism is considered to be one of the major risk factors for osteoporosis in men. However, the mechanisms of bone loss caused by androgen deficiency are still unclear. In the present study, we sequentially investigated the skeletal and hormonal effects of androgen deficiency in aged orchiectomized (ORX) rats over a time period of 9 months. One hundred seventy 13-month-old male Fischer-344 rats were either ORX or sham-operated (SHAM). Eight rats served as baseline controls. After in vivo fluorochrome labeling, groups of 8-15 SHAM and ORX rats each were killed at 2 weeks and at 1, 2, 3, 4, 6, and 9 months postsurgery. As expected, ORX induced a fall in serum total and free testosterone levels, but also reduced serum estradiol concentrations. Cancellous bone area (BAr) in the proximal tibia but not in the first lumbar vertebral body showed an age-dependent decline in SHAM rats. Relative to SHAM controls, ORX rats had significantly reduced cancellous BAr after 2 weeks post-ORX in the tibia and after 2 months post-ORX in the vertebral body. Thereafter, vertebral and tibial cancellous BAr continued to decline in ORX animals throughout the study. Osteoclast number (NOc), osteoblast surface, bone formation rate (BFR), and activation frequency were increased in ORX animals from 1 month postsurgery until the end of the trial. Moreover, in close temporal association with the histomorphometric findings, serum osteocalcin and urinary excretion of collagen cross-links and calcium were elevated in ORX rats. In a stepwise model of multiple regression analysis using estradiol and free and total testosterone as independent variables, estradiol was the only significant predictor of histomorphometric indices of bone formation and bone resorption in SHAM and ORX rats. These data show that androgen deficiency induces substantial loss of cancellous bone in the axial and appendicular skeleton of aged male rats and that this osteopenia is associated with a sustained increase in bone turnover. Thus, the skeletal effects of androgen withdrawal in aged male rats appear to resemble those induced by estrogen withdrawal in female rats. Furthermore, our study suggests that estradiol may act as a physiological suppressor of bone remodeling in aged male rats. PMID- 10841178 TI - Spaceflight effects on cultured embryonic chick bone cells. AB - A model calcifying system of primary osteoblast cell cultures derived from normal embryonic chicken calvaria has been flown aboard the shuttle, Endeavour, during the National Aeronautics and Space Administration (NASA) mission STS-59 (April 9 20, 1994) to characterize unloading and other spaceflight effects on the bone cells. Aliquots of cells (approximately 7 x 10(6)) grown in Dulbecco's modified Eagle's medium (DMEM) + 10% fetal bovine serum (FBS) were mixed with microcarrier beads, inoculated into cartridge culture units of artificial hollow fiber capillaries, and carried on the shuttle. To promote cell differentiation, cartridge media were supplemented with 12.5 microg/ml ascorbate and 10 mM beta glycerophosphate for varying time periods before and during flight. Four cartridges contained cells from 17-day-old embryos grown for 5 days in the presence of ascorbate prior to launch (defined as flight cells committed to the osteoblastic lineage) and four cartridges supported cells from 14-day-old embryos grown for 10 days with ascorbate before launch (uncommitted flight cells). Eight cartridges prepared in the same manner were maintained under normal gravity throughout the flight (control cells) and four additional identical cartridges under normal gravity were terminated on the day of launch (basal cells). From shuttle launch to landing, all cartridges were contained in closed hardware units maintaining 5% CO2, 37 degrees C, and media delivery at a rate of approximately 1.5 ml/6 h. During day 3 and day 5 of flight, duplicate aliquots of conditioned media and accumulated cell products were collected in both the flight and the control hardware units. At the mission end, comparisons among flight, basal, and control samples were made in cell metabolism, gene expression for type I collagen and osteocalcin, and ultrastructure. Both committed and uncommitted flight cells were metabolically active, as measured by glucose uptake and lactate production, at approximately the same statistical levels as control counterparts. Flight cells elaborated a less extensive extracellular matrix, evidenced by a reduced collagen gene expression and collagen protein appearance compared with controls. Osteocalcin was expressed by all cells, a result indicating progressive differentiation of both flight and control osteoblasts, but its message levels also were reduced in flight cells compared with ground samples. This finding suggested that osteoblasts subjected to flight followed a slower progression toward a differentiated function. The summary of data indicates that spaceflight, including microgravity exposure, demonstrably affects bone cells by down regulating type I collagen and osteocalcin gene expression and thereby inhibiting expression of the osteogenic phenotype notably by committed osteoblasts. The information is important for insight into the response of bone cells to changes of gravity and of force in general. PMID- 10841179 TI - Effects of a short-term vitamin D and calcium supplementation on body sway and secondary hyperparathyroidism in elderly women. AB - Long-term vitamin D and calcium supplementation is effective in reducing nonvertebral fractures in elderly people. Increased bone fragility caused by secondary hyperparathyroidism (sHPT) and impaired balance are known risk factors for hip fractures. The hypothesis is that short-term therapy with calcium and vitamin D may improve body sway as well as sHPT more effectively than calcium monotherapy. The effects of 8 weeks of supplementation with vitamin D (cholecalciferol) and calcium on body sway and biochemical measures of bone metabolism were measured. The sample consisted of 148 women (mean [+/-SD] age, 74 +/- 1 years) with a 25-hydroxycholecalciferol level below 50 nmol/liter. They received either 1200 mg of calcium plus 800 IU of vitamin D or 1200 mg of calcium per day. We measured intact parathyroid hormone (PTH), markers of bone turnover, and body sway before and after treatment. Falls and fractures among the participants were followed over a 1-year period. Compared with calcium mono, supplementation with vitamin D and calcium resulted in an increase in serum 25 hydroxyvitamin D of 72% (p < 0.0001), a decrease in the serum PTH of 18% ( p = 0.0432), and a decrease in body sway of 9% (p = 0.0435). The mean number of falls per subject during a 1-year follow-up period was 0.45 for the calcium mono group and 0.24 for the calcium and vitamin D group (p = 0.0346). Short-term supplementation with vitamin D and calcium improves sHPT and body sway and therefore may prevent falls and subsequent nonvertebral fractures in elderly women. PMID- 10841180 TI - Frequency-dependent effect of nitric oxide donor nitroglycerin on bone. AB - Recently, we showed that supplementation with nitric oxide (NO) via donor nitroglycerin (NG) alleviated the ovariectomy and corticosteroid-induced bone loss in rats. In humans, high doses or frequent applications of NG (i.e., for angina) lead to rapid loss of its efficacy in relieving angina. To examine whether there is a similar effect on the loss of efficacy of NG on bone, we examined the frequency-dependent effects of NG on bone mineral density (BMD), bone mass, trabecular bone volumes (BV/TV), and blood pressure in rats. Thirty 7 month-old female Brown Norway rats underwent ovariectomy, and an additional six rats were sham-operated. The ovariectomized rats were treated either with vehicle (ovariectomized control), 17beta-estradiol (E2; positive control), or 0.2 mg NG (via dermal application) once, twice, or three times a day. Before and at the end of the 10-week treatment period, BMD of the lumbar spine was measured by dual energy X-ray absorptiometric (DXA) scanning and expressed as a percentage change. BMD in ovariectomized rats was significantly lower (-2.5 +/- 2.0%) compared with the sham-operated rats (+6.3 +/- 5.3%; p < 0.01). Estrogen therapy completely abolished the ovariectomy-induced potential bone loss (+5.9 +/- 3.4%). Application of NG once daily also completely prevented (+6.2 +/- 2.8%; p < 0.01) the ovariectomy-induced bone loss (i.e., it was as effective as estrogen). However, the beneficial effects of NG on BMD were significantly reduced with increased frequency of application of NG (+1.9 +/- 2.1%, twice a day and -0.2 +/- 3.3% three times a day). Estrogen or once daily administration of NG preserved femur weights, BV/TV, and decreased urinary deoxypyridinoline levels as expected. However, a higher level of serum osteocalcin and bone-specific alkaline phosphatase levels were maintained only with once daily administration of NG. There were no adverse effects of these doses of NG on blood pressure, but a tendency to lower blood pressure was noticed with increased frequency of NG. These results confirmed our previous findings that NO donors counteract the bone loss associated with estrogen deficiency. However, these beneficial effects of maintaining BMD are lost with increased frequency of NG application. PMID- 10841181 TI - Genetic regulation of cortical and trabecular bone strength and microstructure in inbred strains of mice. AB - The inbred strains of mice C57BL/6J (B6) and C3H/HeJ (C3H) have very different femoral peak bone densities and may serve as models for studying the genetic regulation of bone mass. Our objective was to further define the bone biomechanics and microstructure of these two inbred strains. Microarchitecture of the proximal femur, femoral midshaft, and lumbar vertebrae were evaluated in three dimensions using microcomputed tomography (microCT) with an isotropic voxel size of 17 microm. Mineralization of the distal femur was determined using quantitative back-scatter electron (BSE) imaging. MicroCT images suggested that C3H mice had thicker femoral and vertebral cortices compared with B6. The C3H bone tissue also was more highly mineralized. However, C3H mice had few trabeculae in the vertebral bodies, femoral neck, and greater trochanter. The trabecular number (Tb.N) in the C3H vertebral bodies was about half of that in B6 vertebrae (2.8(-1) +/- 0.1 mm(-1) vs. 5.1(-1) +/- 0.2 mm(-1); p < 0.0001). The thick, more highly mineralized femoral cortex of C3H mice resulted in greater bending strength of the femoral diaphysis (62.1 +/- 1.2N vs. 27.4 +/- 0.5N, p < 0.0001). In contrast, strengths of the lumbar vertebra were not significantly different between inbred strains (p = 0.5), presumably because the thicker cortices were combined with inferior trabecular structure in the vertebrae of C3H mice. These results indicate that C3H mice benefit from alleles that enhance femoral strength but paradoxically are deficient in trabecular bone structure in the lumbar vertebrae. PMID- 10841182 TI - Evidence for a major gene for bone mineral density in idiopathic osteoporotic families. AB - Although there have been a number of studies indicating a heritable component for osteoporosis in middle to late adulthood, the etiology of osteoporosis in young people is uncertain. The present study aims to evaluate the extent to which genetic factors influence familial resemblance for bone mineral density (BMD) in families ascertained on the basis of young osteoporotic probands. The sample comprises eight families (74 total individuals) that were identified through a proband under the age of 35 years with a history of two or more fractures and a spinal bone density of at least 2.5 SDs below the mean for age and sex (Z score). Secondary causes of osteoporosis were excluded in the probands. In total, 27% (18/66) of the probands' relatives had osteoporosis and an additional 30% (20/66) had osteopenia. Classical segregation analysis was performed to evaluate the extent to which a genetic etiology could account for familial resemblance in these families. The results indicate a major gene of codominant inheritance for spinal BMD. Model-fitting comparisons revealed no support for environmental effects or for polygenic inheritance. PMID- 10841184 TI - In vivo osteogenic capability of cultured allogeneic bone in porous hydroxyapatite: immunosuppressive and osteogenic potential of FK506 in vivo. AB - Fischer or ACI rat marrow cells were obtained from femoral shafts and were cultured to confluence in Eagle's minimal essential medium (EMEM) supplemented with 15% fetal bovine serum. After trypsinization, the cells were subcultured on porous hydroxyapatite (HA; Interpore 500) blocks in the presence of beta glycerophosphate and 10 nM dexamethasone (Dex). After 2 weeks of subculture, a mineralized bone matrix with osteogenic cells developed on the HA pore surfaces. ACI or Fischer cultured bone tissue/HA constructs were implanted subcutaneously into the backs of Fischer rats and the immunosuppressant FK506 was given to the rats for 4 weeks. Implants were harvested 4 weeks and 8 weeks after insertion. At 4 weeks, the ACI constructs (allografts) showed high levels of osteogenic parameters (alkaline phosphatase [ALP] activity and osteocalcin content) and bone formation was observed together with active osteoblasts without obvious accumulation of inflammatory cells. At 8 weeks, active osteoblasts and progressive bone formation were still observed, while osteogenic parameters remained high and osteocalcin messenger RNA (mRNA) was detected. Without FK506 administration, the allografts showed neither bone formation nor osteocalcin mRNA and there were only trace levels of the osteogenic parameters. In the case of Fischer constructs (isografts), extensive bone formation was detected and all the osteogenic parameters were higher with FK506 than without FK506 at both 4 weeks and 8 weeks. These results indicate that cultured bone tissue/HA constructs possess a high osteogenic potential, even as allografts, and that FK506 not only has an immunosuppressive action, but also promotes bone formation. PMID- 10841183 TI - Involvement of CCAAT enhancer binding protein transcription factors in the regulation of prostaglandin G/H synthase 2 expression by interleukin-1 in osteoblastic MC3T3-E1 cells. AB - Interleukin-1 (IL-1) stimulates prostaglandin production in bone by a rapid and transient activation of prostaglandin G/H synthase 2 (PGHS-2) gene expression. In osteoblastic MC3T3-E1 cells, IL-1 caused a transient increase in PGHS-2 messenger RNA (mRNA), which peaked at 2 h. IL-1 caused a 2- to 4-fold activation of a 371 base pair (bp) murine PGHS-2 promoter/luciferase construct in stable transfectants. This response mapped to a proximal promoter element(s) located between -150 and -40 bp. This region contains a putative CCAAT enhancer binding protein (C/EBP) site (centered at -135 bp), which shows enhanced binding of C/EBPbeta and C/EBPdelta by mobility shift analysis after IL-1 treatment. A transient cotransfection approach was used to examine the effects of C/EBPbeta and C/EBPdelta overexpression. IL-1 caused a maximal 3- to 7-fold stimulation of PGHS-2 promoter activity after 2.5 h. Overexpression of murine C/EBPbeta and C/EBPdelta caused a dose-dependent increase in basal and IL-1-stimulated luciferase activity. C/EBPdelta caused a greater enhancement of basal and IL-1 stimulated promoter activity than C/EBPbeta, suggesting that C/EBPdelta is a stronger transactivator. Overexpression of p20C/EBPbeta, a dominant negative inhibitor of C/EBP function, blocked the stimulation of PGHS-2 promoter activity by IL-1 and blocked the ability of overexpressed C/EBPbeta and C/EBPdelta to increase basal and IL-1-stimulated promoter activity. Mutagenesis of the C/EBP site reduced, but did not abolish, the stimulation of PGHS-2 promoter activity by IL-1 and blunted the effect of overexpressed C/EBPdelta on basal and IL-1 stimulated promoter activity. These results suggest an essential role for C/EBPbeta and C/EBPdelta in the induction of PGHS-2 gene expression by IL-1 in osteoblastic cells. PMID- 10841185 TI - Making rats rise to erect bipedal stance for feeding partially prevented orchidectomy-induced bone loss and added bone to intact rats. AB - The objectives of this study were to investigate the different effects on muscle mass and cancellous (proximal tibial metaphysis [PTM]) and cortical (tibial shaft [TX]) bone mass of sham-operated and orchidectomized (ORX) male rats by making rats rise to erect bipedal stance for feeding. Specially designed raised cages (RC) were used so that the rats had to rise to erect bipedal stance to eat and drink for 12 weeks. Dual-energy X-ray absorptiometry (DEXA) and peripheral quantitative computerized tomography (pQCT) were used to estimate the lean leg mass and bone mineral. Static and dynamic histomorphometry were performed on the triple-labeled undecalcified sections. We found that making the intact rats rise to erect bipedal stance for feeding increased muscle mass, cortical bone volume, and periosteal bone formation. Orchidectomy increased net losses of bone next to the marrow by increasing bone turnover. Making the ORX rats rise to erect bipedal stance increased muscle mass, partially prevented cancellous bone loss in the PTM, and prevented net cortical bone loss in TX induced by ORX by depressing cancellous and endocortical high bone turnover and stimulating periosteal bone formation. The bone-anabolic effects were achieved mainly in the first 4 weeks in the PTM and by 8 weeks in the TX. These findings suggested that making the rats rise to erect bipedal stance for feeding helped to increase muscle mass and cortical bone mass in the tibias of intact rats, increase muscle mass, and partially prevented cancellous and net cortical bone loss in ORX rats. PMID- 10841186 TI - Maturation state determines the response of osteogenic cells to surface roughness and 1,25-dihydroxyvitamin D3. AB - In this study we assessed whether osteogenic cells respond in a differential manner to changes in surface roughness depending on their maturation state. Previous studies using MG63 osteoblast-like cells, hypothesized to be at a relatively immature maturation state, showed that proliferation was inhibited and differentiation (osteocalcin production) was stimulated by culture on titanium (Ti) surfaces of increasing roughness. This effect was further enhanced by 1,25 dihydroxyvitamin D3 [1,25(OH)2D3]. In the present study, we examined the response of three additional cell lines at three different maturation states: fetal rat calvarial (FRC) cells (a mixture of multipotent mesenchymal cells, osteoprogenitor cells, and early committed osteoblasts), OCT-1 cells (well differentiated secretory osteoblast-like cells isolated from calvaria), and MLO Y4 cells (osteocyte-like cells). Both OCT-1 and MLO-Y4 cells were derived from transgenic mice transformed with the SV40 large T-antigen driven by the osteocalcin promoter. Cells were cultured on Ti disks with three different average surface roughnesses (Ra): PT, 0.5 microm; SLA, 4.1 microm; and TPS, 4.9 microm. When cultures reached confluence on plastic, vehicle or 10(-7) M or 10( 8) M 1,25(OH)2D3 was added for 24 h to all of the cultures. At harvest, cell number, alkaline phosphatase-specific activity, and production of osteocalcin, transforming growth factor beta1 (TGF-beta1) and prostaglandin E2 (PGE2) were measured. Cell behavior was sensitive to surface roughness and depended on the maturation state of the cell line. Fetal rat calvarial (FRC) cell number and alkaline phosphatase-specific activity were decreased, whereas production of osteocalcin, TGF-beta1, and PGE2 were increased with increasing surface roughness. Addition of 1,25(OH)2D3 to the cultures further augmented the effect of roughness for all parameters in a dose-dependent manner; only TGF-beta1 production on plastic and PT was unaffected by 1,25(OH)2D3. OCT-1 cell number and alkaline phosphatase (SLA > TPS) were decreased and production of PGE2, osteocalcin, and TGF-beta1 were increased on SLA and TPS. Response to 1,25(OH)2D3 varied with the parameter being measured. Addition of the hormone to the cultures had no effect on cell number or TGF-beta1 production on any surface, while alkaline phosphatase was stimulated on SLA and TPS; osteocalcin production was increased on all Ti surfaces but not on plastic; and PGE2 was decreased on plastic and PT, but unaffected on SLA and TPS. In MLO-Y4 cultures, cell number was decreased on SLA and TPS; alkaline phosphatase was unaffected by increasing surface roughness; and production of osteocalcin, TGF-beta1, and PGE2 were increased on SLA and TPS. Although 1,25(OH)2D3 had no effect on cell number, alkaline phosphatase, or production of TGF-beta1 or PGE2 on any surface, the production of osteocalcin was stimulated by 1,25(OH)2D3 on SLA and TPS. These results indicate that surface roughness promotes osteogenic differentiation of less mature cells, enhancing their responsiveness to 1,25(OH)2D3. As cells become more mature, they exhibit a reduced sensitivity to their substrate but even the terminally differentiated osteocyte is affected by changes in surface roughness. PMID- 10841187 TI - Analysis of proximal femur DXA scans in growing children: comparisons of different protocols for cross-sectional 8-month and 7-year longitudinal data. AB - Dual-energy X-ray absorptiometry (DXA) is a widely used method for measuring bone mineral in the growing skeleton. Because scan analysis in children offers a number of challenges, we compared DXA results using six analysis methods at the total proximal femur (PF) and five methods at the femoral neck (FN). In total we assessed 50 scans (25 boys, 25 girls) from two separate studies for cross sectional differences in bone area, bone mineral content (BMC), and areal bone mineral density (aBMD) and for percentage change over the short term (8 months) and long term (7 years). At the proximal femur for the short-term longitudinal analysis, there was an approximate 3.5% greater change in bone area and BMC when the global region of interest (ROI) was allowed to increase in size between years as compared with when the global ROI was held constant. Trend analysis showed a significant (p < 0.05) difference between scan analysis methods for bone area and BMC across 7 years. At the femoral neck, cross-sectional analysis using a narrower (from default) ROI, without change in location, resulted in a 12.9 and 12.6% smaller bone area and BMC, respectively (both p < 0.001). Changes in FN area and BMC over 8 months were significantly greater (2.3%, p < 0.05) using a narrower FN rather than the default ROI. Similarly, the 7-year longitudinal data revealed that differences between scan analysis methods were greatest when the narrower FN ROI was maintained across all years (p < 0.001). For aBMD there were no significant differences in group means between analysis methods at either the PF or FN. Our findings show the need to standardize the analysis of proximal femur DXA scans in growing children. PMID- 10841188 TI - Molecular events that contribute to lysyl oxidase enzyme activity and insoluble collagen accumulation in osteosarcoma cell clones. AB - Maximum collagen synthesis and maximum accumulation of insoluble collagen occur at different phenotypic stages in developing osteoblastic cell cultures. Insoluble collagen accumulation depends in part on the activity of extracellular enzymes including procollagen N-proteinases, procollagen C-proteinase (derived from the BMP1 gene), and lysyl oxidase. In addition to its action on procollagen, procollagen C-proteinase processes prolysyl oxidase to mature 32-kDa lysyl oxidase. The regulation of extracellular activities that control insoluble collagen accumulation has not been studied extensively. The present study compares molecular events that control production of a collagenous mineralized extracellular matrix in vitro among five different murine osteosarcoma cell clones derived from the same tumor, but which differ in their ability to produce an insoluble mineralized matrix. Levels of insoluble type I collagen, insoluble calcium, bone morphogenetic protein 1 (BMP-1), and lysyl oxidase expression, lysyl oxidase biosynthesis, lysyl oxidase activity, and prolysyl oxidase processing activity were determined. Results surprisingly indicate that lysyl oxidase activity is not related closely to lysyl oxidase messenger RNA (mRNA) levels among the different cell clones. However, it appears that BMP-1-dependent prolysyl oxidase processing could contribute to the observed lysyl oxidase activity. Highest collagen and BMP-1 mRNA levels, prolysyl oxidase processing activity, and lysyl oxidase activity occurred in a cell clone (K8) that showed the highest levels of insoluble collagen accumulation. Culture media from a cell clone (K37) that accumulates little insoluble collagen or calcium but expresses high levels of lysyl oxidase mRNA contained low molecular weight fragments of lysyl oxidase protein and showed low lysyl oxidase activity. By contrast the K14 cell line exhibits relatively high lysyl oxidase activity and collagen accumulation, but low levels of mature lysyl oxidase protein. Together, these studies indicate that catabolic as well as anabolic activities are important in regulating insoluble collagen accumulation in osteoblastic cells. In addition, results suggest that products of genes homologous to lysyl oxidase may contribute to observed lysyl oxidase activity. PMID- 10841189 TI - Local drug delivery. AB - Intensive research efforts are now focused on the development of new strategies for more effective delivery of drugs to the central nervous system. These strategies include chemical modification of drugs, disruption of the blood-brain barrier, and utilization of alternative routes for drug delivery. This paper focuses on local drug delivery for the treatment of brain tumors. It reviews papers published in the past year on local chemotherapy and immunotherapy. Other aspects of local drug delivery are discussed, including convection-enhanced delivery and drug delivery via a controlled-release microchip. PMID- 10841190 TI - Treatment of low-grade pediatric gliomas. AB - The prognosis of pediatric low-grade gliomas is relatively favorable, but residual or unresectable tumors remain a challenge. Conventional radiation therapy is effective but may be associated with unacceptable sequelae in young children. Chemotherapy can delay the need for radiation therapy in young children; however, its role in older children has yet to be established. New forms of radiation are being introduced that may be less toxic. PMID- 10841191 TI - Magnetic resonance spectroscopy of brain tumors. AB - Magnetic resonance spectroscopy provides metabolic information about brain tumors beyond what can be obtained from anatomic images. In contrast to other metabolism based imaging techniques such as single photon emission computed tomography and positron-emission tomography, magnetic resonance spectroscopy yields multiparametric data, does not require radio-labeled tracers or ionizing radiation, and can be performed in conjunction with other magnetic resonance imaging studies. Magnetic resonance spectral patterns have been shown to be distinct for different tumor types and grades. Response to radiation therapy is also reflected by magnetic resonance spectral patterns. Although there are quantitative issues still to be addressed, correlation of in vivo spectral patterns with ex vivo spectral patterns obtained from actual biopsy samples indicates that magnetic resonance spectroscopy is a fundamentally valid tool for monitoring disease progression and therapeutic response in patients with brain tumors. PMID- 10841192 TI - Altered fractionation in definitive irradiation of squamous cell carcinoma of the head and neck. AB - The likelihood of local control after radiation therapy may be improved by increasing total dose or decreasing overall time. The probability of late complications increases with dose per fraction. Altered fractionation techniques usually employ two or more fractions per day using a dose per fraction that is similar or less than that employed in conventional fractionation. Altered fractionation may be broadly classified as hyperfractionation or accelerated fractionation. Data suggest that altered fractionation schedules may improve local control (and to a lesser extent, survival) compared with conventional irradiation. PMID- 10841193 TI - Evaluation of quality of life in patients definitively treated for squamous carcinoma of the head and neck. AB - This paper reviews quality-of-life (QOL) assessment in head and neck cancer with a focus on research and methodologic developments of the past year. Issues of QOL measurement, as well as the challenges to data synthesis in light of the heterogeneity of head and neck cancer and its treatment, are discussed briefly. Methodologic advances include increased use of validated measures, longitudinal study design, and attention to patients' attitudes. Although the majority of studies assessed multiple QOL domains, including both physical/functional and emotional/social, more focused areas of investigation included pain, organ preservation, and depression. Examinations of the relation among domains suggested that impaired function does not necessarily lead to poor QOL and that the best predictor of 12-month global QOL is pretreatment global QOL. Future challenges include the need for large multi-institutional studies, consensus about instrument selection, addressing the problem of missing data, and how to apply group OOL to individual patient decisions. PMID- 10841194 TI - Integration of taxanes into primary chemotherapy for squamous cell carcinoma of the head and neck: promise fulfilled? AB - Chemotherapy has become integrated into the treatment of head and neck cancer in not only the palliative but now also the primary setting. Organ preservation is possible using induction chemotherapy, and improved survival results have been confirmed for concomitant chemoradiotherapy. The taxanes, paclitaxel and docetaxel, appear to be as active as any other drugs in head and neck cancer. When used in combination in the induction, recurrent, or metastatic settings, response rates rival those of the standard cisplatin/5-fluorouracil regimen. At least one ongoing study will help to establish superiority of cisplatin/paclitaxel versus cisplatin/5-fluorouracil in the recurrent or metastatic setting, and another between cisplatin/5-fluorouracil and doctaxel/cisplatin/5-fluorouracil in the induction setting. Both paclitaxel and docetaxel are being extensively studied as radiosensitizers. They are relatively well tolerated and have good efficacy but have not yet been adequately studied in comparison with other regimens. In conclusion, the taxanes have significantly expanded our effective treatment options in both the primary and recurrent or metastatic settings. PMID- 10841195 TI - Molecular markers predictive of response and prognosis in the patient with advanced squamous cell carcinoma of the head and neck: evolution of a model beyond TNM staging. AB - Advanced head and neck cancer is a disease with poor prognosis. TNM staging is an inadequate prognostic indicator of individual response to evolving multimodal therapies. New markers have been studied in progressively more refined analyses. Even though their role in predicting response and prognosis of head and neck cancer is still under evolution, it is becoming clear that individual markers are inadequate in constructing a prognostically meaningful tumor profile for each patient. Rather the combined study of a number of well-characterized markers acting in unrelated cellular pathways may be much more successful in defining prognostic patient categories of greater utility than traditional TNM staging. Special attention should also be paid to the expression pattern and location of tumor markers within the biopsy specimen as these parameters also appear to influence prognostic significance. PMID- 10841196 TI - Carcinogenesis of head and neck cancer and the role of chemoprevention in its reversal. AB - Head and neck cancer is an important public health problem worldwide, accounting for approximately 40,400 new cancer cases and 12,300 cancer deaths annually in the US. Although early-stage disease is often curable with surgery or radiotherapy, the majority of patients present with advanced disease in which despite advances in combined modality therapy the outcomes have not dramatically improved. Furthermore, patients cured of their initial early-stage head and neck squamous cell carcinoma are at high risk for development of second primary tumors, which pose the main threat to survival. An alternative approach in reducing the incidence and thus mortality associated with these cancers is chemoprevention, the use of agents to reverse, halt, or delay carcinogenesis. The carcinogenesis process in head and neck cancer results from a dysregulation of cellular proliferation, differentiation, and cell death resulting from field-wide exposure of the upper aerodigestive tract to tobacco smoking. Newly acquired knowledge in the field of tumor biology and of the genetic changes underlying carcinogenesis through the use of new molecular technology represents the basis on which chemoprevention efforts should be based. PMID- 10841197 TI - Current opinion in germ cell cancer 2000. AB - Despite its relative rarity compared with the common adult cancers, scientific and clinical interest in germ cell cancer is increasing. From the point of view of epidemiology, the controversy about the relative importance of intrauterine versus postpubertal risk factors has continued. Evidence to support the importance of intrauterine factors comes from reports from Norway, Canada, and the US, confirming the Danish observation that the rising incidence of germ cell cancer is linked to a birth cohort effect; evidence in support of the importance of postpubertal risk comes from three case/control studies demonstrating increased risk linked to postpubertal exposures such as pesticides, plastics, electromagnetic radiation, trauma, and infections. There has been increasing interest in human endogenous retrovirus K10 as a possible factor explaining genetic susceptibility and providing a linkage between the two groups of risk factors. In cytogenetics, progress was reported in identifying the deletion point of the suspected tumor suppressor gene responsible for the i12p marker chromosome abnormality and development of FISH probes for diagnostic purposes. In molecular biology, the importance of DNA repair deficiency in normal germ cells as a factor in the exquisite chemosensitivity of germ cell cancer has been high-lighted by a report demonstrating a low level of the xeroderma pigmentosa group A (XPA) protein and induction of resistance in vitro by adding XPA. In the clinic, progress in positron emission tomography scanning and laparoscopic lymph node staging are leading to changes in outlook on management of stage 1 cases and patients with small residual masses postchemotherapy. Salvage chemotherapy regimens integrating dose dense and vertical dose intensification strategies reported 60% progression-free survival. New drugs such as gemcitabine demonstrated continued therapeutic potential for chemotherapy in these tumors. A report demonstrating the inadequacies of hormone replacement after bilateral orchidectomy and a report of the first child born after testis-conserving therapy highlight the need for more attention to testis conservation as a quality of life issue. With the cure rates so high, the need for central referral is once again debated both for stage 1 and metastatic disease. With new ways of defining poor risk stage 1 patients and reports on impact of experience highlighting the worse outcome of patients treated in centers treating small numbers, views on this issue remain clearcut. PMID- 10841198 TI - Bladder cancer. AB - There is a need for the development of reliable tumor markers in bladder cancer. A number of studies this past year focused on the evaluation of urinary markers that hold promise as noninvasive adjuncts to traditional diagnostic or surveillance techniques, principally urinary cytology and cystoscopy. Tests for bladder tumor antigen, NMP22, and fibrin degradation products, as well as the Immunocyt test, are commercially available. Other urinary marker tests discussed in this review include telomerase, cytokeratins, and vascular endothelial growth factor. Although these tests in many instances have improved sensitivity in detecting bladder cancer compared with urinary cytology, none have become widely accepted in routine clinical practice. Nonetheless, with further refinement and prospective validation in multicenter trials, markers such as these may provide information that would permit tailoring on an individual basis the type of as well as interval of surveillance examinations. Furthermore, they may also provide information allowing the appropriate selection of therapy based on predicted response. In addition to urinary markers, intense research efforts have also focused on developing clinically useful molecular prognostic markers. A number of cell-cycle regulatory proteins, including p53 and p21, have received much attention in this regard. Emerging data suggests that it may soon be possible to determine the molecular phenotype of both superficial and invasive bladder cancers, thereby providing information regarding tumor behavior on an individual basis. As with urinary markers, however, no molecular markers have been incorporated as yet into day-to-day patient care. Assurances of reproducibility, standardization, and prospective validation studies are urgently needed. It is only through this type of rigorous evaluation that the level of confidence sufficient to base treatment decisions on marker status will be attained. PMID- 10841199 TI - Renal cell carcinoma. AB - The overall incidence of renal cell carcinoma is rising, for reasons not fully explained by increased abdominal imaging. Risk factors associated with renal cell carcinoma include hypertension, smoking, increased body mass index, and diet. There is an inverse association of renal cell carcinoma risk with consumption of a variety of carotenes. In addition, increased red meat intake has been associated with increased risk. Partial nephrectomy may be as effective as radical nephrectomy as treatment for localized disease, and radiosurgery may be as effective as surgical resection in the management of brain metastases. Immunotherapy remains the mainstay for systemic treatment, with response rates between 5% and 20%. Survival in renal cell carcinoma is related to pathologic stage, nuclear grade, microscopic vascular invasion, DNA content, nuclear morphometry, and histologic pattern. In addition, patients with deletion (8p)/-8, +12, and +20 appear to have a worse prognosis. PMID- 10841200 TI - An update on prostate cancer research. AB - The pathogenesis of prostate cancer reflects complex interactions among environmental and genetic factors. Recent advances suggest molecular mechanisms that may explain geographic and ethnic variations in prostate cancer incidence, and understanding of molecular disease progression is advancing rapidly. Clinically, the case for screening has become stronger, and declining prostate cancer mortality rates may be due in part to early detection and treatment. Improved risk assessment for patients with localized disease is now available, although further refinement in predictive algorithms will need to incorporate validated molecular prognostic markers. Treatment options for patients with localized prostate cancer have expanded and the role of androgen deprivation further delineated. Finally, treatment strategies for patients with androgen independent disease have also expanded, although novel therapies are required to improve survival in this group of patients. PMID- 10841201 TI - Pediatric genitourinary tumors. AB - Advances in our knowledge of pediatric genitourinary tumors are being made at both the basic science and clinical levels. The molecular mechanisms underlying these pediatric malignancies are being uncovered and will aid in uncovering novel treatments. Because of the high success rate in treating these tumors, treatment options are being modified to decrease both short- and long-term morbidity, while maintaining the improved survival. PMID- 10841202 TI - Bibliography. Current world literature. Brain and nervous system. PMID- 10841203 TI - Bibliography. Current world literature. Head and neck. PMID- 10841204 TI - Bibliography. Current world literature. Genitourinary systems. PMID- 10841205 TI - Circadian rhythms and sleep in human aging. AB - This issue of Chronobiology International is dedicated to the age-related changes in circadian rhythms as they occur in humans. It seems timely to give an overview of the knowledge and hypotheses on these changes now that we enter a century in which the number and percentage of elderly in the population will be unprecedented. Although we should take care not to follow the current tendency to think of old age as a disease--ignoring the fine aspects of being old--there is definitely an age-related increase in the risk of a number of conditions that are at least uncomfortable. Circadian rhythms have been attributed adaptive values that usually go unnoticed, but can surface painfully clear when derangements occur. Alterations in the regulation of circadian rhythms are thought to contribute to the symptoms of a number of conditions for which the risk is increased in old age (e.g., sleep disturbances, dementia, and depression). A multidisciplinary approach to investigate the mechanisms of age-related changes in circadian regulation eventually may result in treatment strategies that will improve the quality of life of the growing number of elderly. Although diverse topics are addressed in this issue, the possible mechanisms by which a deranged circadian timing system may be involved in sleep disturbances receives the most attention. This seems appropriate in view of the numerous studies that have addressed this relation in the last decade and also because of the high frequency and strong impact of sleep disturbances in the elderly. This introduction to the special issue first briefly addresses the impact of disturbed sleep in the elderly to show that the development of therapeutic methods other than the currently available pharmacological treatments should be given high priority. I believe that chronobiological insights may play an important role in the development of rational therapeutical methods. PMID- 10841206 TI - The human circadian clock and aging. AB - The suprachiasmatic nucleus (SCN) of the hypothalamus is implicated in the timing of a wide variety of circadian processes. Since the environmental light-dark cycle is the main zeitgeber for many of the rhythms, photic information may have a synchronizing effect on the endogenous clock of the SCN by inducing periodic changes in the biological activity of certain groups of neurons. By studying the brains obtained at autopsy of human subjects, marked diurnal oscillations were observed in the neuropeptide content of the SCN. Vasopressin, for example, one of the most abundant peptides in the human SCN, exhibited a diurnal rhythm, with low values at night and peak values during the early morning. However, with advancing age, these diurnal fluctuations deteriorated, leading to a disrupted cycle with a reduced amplitude in elderly people. These findings suggest that the synthesis of some peptides in the human SCN exhibits an endogenous circadian rhythmicity, and that the temporal organization of these rhythms becomes progressively disturbed in senescence. PMID- 10841207 TI - Age-dependent changes of the circadian system. AB - This review summarizes the current knowledge on changes of the circadian system in advanced age, mainly for rodents. The first part is dedicated to changes of the overt rhythms. Possible causes are discussed, as are methods to treat the disturbances. In aging animals and humans, all rhythm characters change. The most prominent changes are the decrease of the amplitude and the diminished ability to synchronize with a periodic environment. The susceptibility to photic and nonphotic cues is decreased. As a consequence, both internal and external temporal order are disturbed under steady-state conditions and, even more, following changes in the periodic environment. Due to the high complexity of the circadian system, which includes oscillator(s), mechanisms of external synchronization and of internal coupling, the changes may arise for several reasons. Many of the changes seem to occur within the SCN itself. The number of functioning neurons decreases with advancing age and, probably, so does the coupling between them. As a result, the SCN is unable, or at least less able, to produce stable rhythms and to transmit timing information to target sites. Initially, only the ability to synchronize with the periodic environment is diminished, whereas the rhythms themselves continue to be well pronounced. Therefore, the possibility exists to treat age-dependent disturbances. This can be done pharmacologically or by increasing the zeitgeber strength. So, some of the rhythm disturbances can be reversed, increasing the magnitude of the light dark (LD) zeitgeber. Another possibility is to strengthen feedback effects, for example, by increasing the daily amount of activity. By this means, the stability and synchronization of the circadian activity rhythm of old mice and men were improved. PMID- 10841208 TI - Contribution of circadian physiology and sleep homeostasis to age-related changes in human sleep. AB - The circadian pacemaker and sleep homeostasis play pivotal roles in vigilance state control. It has been hypothesized that age-related changes in the human circadian pacemaker, as well as sleep homeostatic mechanisms, contribute to the hallmarks of age-related changes in sleep, that is, earlier wake time and reduced sleep consolidation. Assessments of circadian parameters in healthy young (approximately 20-30 years old) and older people (approximately 65-75 years old)- in the absence of the confounding effects of sleep, changes in posture, and light exposure--have demonstrated that an earlier wake time in older people is accompanied by about a 1 h advance of the rhythms of core body temperature and melatonin. In addition, older people wake up at an earlier circadian phase of the body temperature and plasma melatonin rhythm. The amplitude of the endogenous circadian component of the core body temperature rhythm assessed during constant routine and forced desynchrony protocols is reduced by 20-30% in older people. Recent assessments of the intrinsic period of the human circadian pacemaker in the absence of the confounding effects of light revealed no age-related reduction of this parameter in both sighted and blind individuals. Wake maintenance and sleep initiation are not markedly affected by age except that sleep latencies are longer in older people when sleep initiation is attempted in the early morning. In contrast, major age-related reductions in the consolidation and duration of sleep occur at all circadian phases. Sleep of older people is particularly disrupted when scheduled on the rising limb of the temperature rhythm, indicating that the sleep of older people is more susceptible to arousal signals generated by the circadian pacemaker. Sleep-homeostatic mechanisms, as assayed by the sleep deprivation-induced increase of EEG slow-wave activity (SWA), are operative in older people, although during both baseline sleep and recovery sleep SWA in older people remains at lower levels. The internal circadian phase advance of awakening, as well as the age-related reduction in sleep consolidation, appears related to an age-related reduction in the promotion of sleep by the circadian pacemaker during the biological night in combination with a reduced homeostatic pressure for sleep. Early morning light exposure associated with this advance of awakening in older people could reinforce the advanced circadian phase. Quantification of the interaction between sleep homeostasis and circadian rhythmicity contributes to understanding age-related changes in sleep timing and quality. PMID- 10841209 TI - More than a marker: interaction between the circadian regulation of temperature and sleep, age-related changes, and treatment possibilities. AB - The neurobiological mechanisms of both sleep and circadian regulation have been unraveled partly in the last decades. A network of brain structures, rather than a single locus, is involved in arousal state regulation, whereas the suprachiasmatic nucleus (SCN) has been recognized as a key structure for the regulation of circadian rhythms. Although most models of sleep regulation include a circadian component, the actual mechanism by which the circadian timing system promotes--in addition to homeostatic pressure--transitions between sleep and wakefulness remains to be elucidated. Little more can be stated presently than a probable involvement of neuronal projections and neurohumoral factors originating in the SCN. This paper reviews the relation among body temperature, arousal state, and the circadian timing system and proposes that the circadian temperature rhythm provides an additional signaling pathway for the circadian modulation of sleep and wakefulness. A review of the literature shows that increased brain temperature is associated with a type of neuronal activation typical of sleep in some structures (hypothalamus, basal forebrain), but typical of wakefulness in others (midbrain reticular formation, thalamus). Not only local temperature, but also skin temperature are related to the activation type in these structures. Warming of the skin is associated with an activation type typical of sleep in the midbrain reticular formation, hypothalamus, and cerebral cortex (CC). The decreasing part of the circadian rhythm in core temperature is mainly determined by heat loss from the skin of the extremities, which is associated with strongly increased skin temperature. As such, alterations in core and skin temperature over the day could modulate the neuronal activation state or "preparedness for sleep" in arousal-related brain structures. Body temperature may thus provide a third signaling pathway, in addition to synaptic and neurohumoral pathways, for the circadian modulation of sleep. A proposed model for the effects of body temperature on sleep appears to fit the available data better than previous hypotheses on the relation between temperature and sleep. Moreover, when the effects of age-related thermoregulatory alterations are introduced into the model, it provides an adequate description of age-related changes in sleep, including shallow sleep and awakening closer to the nocturnal core temperature minimum. Finally, the model indicates that appropriately timed direct (passive heating) or indirect (bright light, melatonin, physical activity) manipulation of the nocturnal profile of skin and core temperature may be beneficial to disturbed sleep in the elderly. Although such procedures could be viewed by researchers as merely masking a marker for the endogenous rhythm, they may in fact be crucial for sleep improvement in elderly subjects. PMID- 10841210 TI - Circadian rhythms in healthy aging--effects downstream from the pacemaker. AB - Using both previously published findings and entirely new data, we present evidence in support of the argument that the circadian dysfunction of advancing age in the healthy human is primarily one of failing to transduce the circadian signal from the circadian timing system (CTS) to rhythms "downstream" from the pacemaker rather than one of failing to generate the circadian signal itself. Two downstream rhythms are considered: subjective alertness and objective performance. For subjective alertness, we show that in both normal nychthemeral (24 h routine, sleeping at night) and unmasking (36 h of constant wakeful bed rest) conditions, advancing age, especially in men, leads to flattening of subjective alertness rhythms, even when circadian temperature rhythms are relatively robust. For objective performance, an unmasking experiment involving manual dexterity, visual search, and visual vigilance tasks was used to demonstrate that the relationship between temperature and performance is strong in the young, but not in older subjects (and especially not in older men). PMID- 10841211 TI - Alterations with aging of the endocrine and neuroendocrine circadian system in humans. PMID- 10841212 TI - Sleep impairments in healthy seniors: roles of stress, cortisol, and interleukin 1 beta. AB - STUDY OBJECTIVES: Increased stress responsivity and a longer-lasting glucocorticoid increase are common findings in aging studies. Increased cortisol levels at the circadian nadir also accompany aging. We used 24 h free urine cortisol to assess these age changes in healthy seniors. We hypothesized that free cortisol levels would explain individual differences in age-related sleep impairments. DESIGN: The study compared sleep, cortisol, and sleep-cortisol correlations under baseline and "stress" conditions in men and women. SETTING: Subjects were studied in the General Clinical Research Center under baseline conditions and a mildly stressful procedure (24 h indwelling intravenous catheter placement). PARTICIPANTS: Eighty-eight healthy, nonobese subjects (60 women and 28 men) from a large study of successful aging participated in the study. Mean ages were 70.6 (+/-6.2) and 72.3 (+/-5.7) years for women and men, respectively. MEASUREMENTS: The 24 h urines were collected for cortisol assay (radioimmunoassay [RIA]); blood was sampled at three diurnal time points for assay (enzyme-linked immunosorbent assay [ELISA]) of interleukin-1 (IL-1) beta; sleep architecture and sleep electroencephalograms (EEGs) were analyzed (after an adaptation and screening night) on baseline and stress nights via polysomnography and EEG power spectral analysis. RESULTS: Healthy older women and men with higher levels of free cortisol (24 h urine level) under a mild stress condition had impaired sleep (lower sleep efficiency; fewer minutes of stages 2, 3, and 4 sleep; more EEG beta activity during non-rapid eye movement sleep [NREM] sleep). Similar results were obtained when stress reactivity measures were used (cortisol and sleep values adjusted for baseline values), but not when baseline values alone were used. Gender differences were apparent: Men had higher levels of free urine cortisol in both baseline and mild stress conditions. Cortisol and sleep correlated most strongly in men; cortisol stress response levels explained 36% of the variance in NREM sleep stress responses. In women, but not men, higher cortisol was also associated with earlier time of arising and less REM sleep. Higher cortisol response to stress was associated with increased circulating levels of IL-1beta, explaining 24% of the variance in a subset of women. CONCLUSION: These results indicate that free cortisol (as indexed by 24 h urine values) can index responses to mild stress in healthy senior adults, revealing functional correlations (impaired sleep, earlier times of arising, more EEG beta activity during sleep, more IL-1beta) and gender differences. PMID- 10841213 TI - Circadian rhythms of agitation in institutionalized patients with Alzheimer's disease. AB - Agitation is a common problem in institutionalized patients with Alzheimer's disease (AD). "Sundowning," or agitation that occurs primarily in the evening, is estimated to occur in 10-25% of nursing home patients. The current study examined circadian patterns of agitation in 85 patients with AD living in nursing homes in the San Diego, California, area. Agitation was assessed using behavioral ratings collected every 15 minutes over 3 days, and activity and light exposure data were collected continuously using Actillume recorders. A five-parameter extension of the traditional cosine function was used to describe the circadian rhythms. The mean acrophase for agitation was 14:38, although there was considerable variability in the agitation rhythms displayed by the patients. Agitation rhythms were more robust than activity rhythms. Surprisingly, only 2 patients (2.4%) were "sundowners." In general, patients were exposed to very low levels of illumination, with higher illumination during the night being associated with less robust agitation rhythms with higher rhythm minima (i.e., some agitation present throughout the day and night). Seasonality was examined; however, there were no consistent seasonal patterns found. This is the largest study to date to examine agitation rhythms using behavioral observations over multiple 24 h periods. The results suggest that, although sundowning is uncommon, agitation appears to have a strong circadian component in most patients that is related to light exposure, sleep, and medication use. Further research into the understanding of agitation rhythms is needed to examine the potential effects of interventions targeting sleep and circadian rhythms. PMID- 10841214 TI - Supplementary administration of artificial bright light and melatonin as potent treatment for disorganized circadian rest-activity and dysfunctional autonomic and neuroendocrine systems in institutionalized demented elderly persons. AB - Increased daytime napping, early morning awakening, frequent nocturnal sleep interruptions, and lowered amplitude and phase advance of the circadian sleep wake rhythm are characteristic features of sleep-waking and chronobiological changes associated with aging. Especially in elderly patients with dementia, severely fragmented sleep-waking patterns are observed frequently and are associated with disorganized circadian rhythm of various physiological functions. Functional and/or organic deterioration of the suprachiasmatic nucleus (SCN), decreased exposure to time cues such as insufficient social interaction and reduced environmental light, lowered sensitivity of sensory organs to time cues, and reduced ability of peripheral effector organs to express circadian rhythms may cause these chronobiological changes. In many cases of dementia, the usual treatments for insomnia do not work well, and the development of an effective therapy is an important concern for health care practitioner and researchers. Recent therapeutical trials of supplementary administration of artificial bright light and the pineal hormone melatonin, a potent synchronizer for mammalian circadian rhythm, have indicated that these treatments are useful tools for demented elderly insomniacs. Both bright light and melatonin simultaneously ameliorate disorganized thermoregulatory and neuroendocrine systems associated with disrupted sleep-waking times, suggesting a new, potent therapeutic means for insomnia in the demented elderly. Future studies should address the most effective therapeutic design and the most suitable types of symptoms for treatment and investigate the use of these tools in preventive applications in persons in early stages of dementia. PMID- 10841215 TI - Low molecular weight heparin in acute coronary syndrome: evidence for superior or equivalent efficacy compared with unfractionated heparin? AB - This article will review the results of recent clinical trials evaluating low molecular weight heparins (LMWHs) in the management of patients with acute coronary syndromes of unstable angina and non-ST segment elevation MI. Low molecular weight heparins are a new class of anticoagulants that have a number of advantages over unfractionated heparin (UFH) leading to their increasing use for thrombotic vascular disorders. There is convincing evidence that LMWH is more effective than placebo and at least as effective as UFH in reducing the hard end points of death and recurrent myocardial infarction. Convincing evidence for a superior efficacy is mostly limited to the least robust but most prevalent end point of recurrent angina, and benefits appear to be confined predominantly to high-risk patients. The benefits are sustained long-term, but there appears to be no incremental benefit with prolonged treatment. The risk for major bleeding is approximately equivalent to UFH, but minor hemorrhage is clearly increased, especially with vascular instrumentation. The increased bleeding risk together with its long half-life and absence of specific antidote warrants exercising caution when using LMWH with coronary intervention. Low molecular weight heparins have the potential of being cost-neutral or even cost-saving by reducing resource utilization, especially in the setting of aggressive interventional practice pattern. Last, the issue of whether one LMWH preparation is more effective and cost-effective than others remains an open question that can be answered only by direct head-to-head comparison of different LMWH preparations in randomized trials. In conclusion, subcutaneous weight-adjusted LMWH is as effective and safe as intravenous UFH in the management of patients with acute coronary syndromes. The logistic ease of administration without the need for monitoring anticoagulation appears to be the major advantage over UFH. PMID- 10841216 TI - Corrected coronary flow velocity reserve: a new concept for assessing coronary perfusion. AB - OBJECTIVES: In order to limit the variability of coronary flow velocity reserve (CFVR), we analyzed which factors independently affect CFVR and established a new parameter integrating these factors. BACKGROUND: Coronary flow velocity reserve (CFVR) is a frequently used parameter for evaluating the physiological significance of epicardial stenosis and microvascular function. Since CFVR measurements are done in substantially different hemodynamic and clinical situations, interpretation of CFVR requires correction for major influencing factors. METHODS: In 141 patients with angina-like symptoms and angiographically unobstructed coronary arteries, intracoronary Doppler measurements were performed in at least two coronary vessels. Coronary flow velocity reserve was calculated as the ratio of hyperemic average peak velocity (hAPV), after intracoronary bolus of adenosine, to baseline average peak velocity (bAPV). RESULTS: Analysis of covariance revealed that only bAPV (p < 0.0001) and age (p < 0.0001) were independent factors influencing CFVR. Based on a regression model for estimation of predicted CFVR values, individual CFVR values (CFVRind) obtained at different bAPV and age were transformed in corrected CFVR values (CFVRcorr) by relating them to a mean bAPV of 15 cm/s and a mean age of 55 years. The transformation from CFVRind into CFVRcorr for the left anterior descending artery can be done by using the following equation: CFVRcorr = 2.85*CFVR(ind)*10(0.48*log(bAPV)+(0.0025*age)-1.16). When applying this new parameter to conditions assumed to cause microvascular dysfunction, analysis showed that only patients with diabetes showed a significant decrease of traditional CFVR and CFVRcorr, whereas a history of hypertension and current smoking habit had no influence on CFVRcorr. CONCLUSIONS: The concept of CFVRcorr standardizes CFVR for bAPV and age as the major physiological determinants. Especially in patients with microvascular dysfunction, this approach may help to discriminate between conditions directly affecting vasodilator reserve and conditions primarily affecting bAPV. PMID- 10841217 TI - Safety and efficacy of elective carotid artery stenting in high-risk patients. AB - OBJECTIVES: We sought to evaluate the safety and efficacy of carotid artery stenting (CAS) in high risk patients. BACKGROUND: Carotid endarterectomy (CE) has been shown to be more effective than medical therapy, but it has limitations. Carotid artery stenting may be a reasonable alternative, particularly in high risk patients. METHODS: We prospectively evaluated the safety and efficacy of CAS in 170 consecutive patients who underwent the procedure in 192 carotid arteries. Of the patients enrolled, 129 (76%) would have been excluded from the major trials of CE and 54 (32%) were referred by vascular surgeons. This series represents a very high-risk group that included patients with unstable angina, previous ipsilateral CE, contralateral carotid artery occlusion and other severe comorbid illnesses. Only 25 (24%) of 104 symptomatic patients would have met the North American Symptomatic Carotid Endarterectomy Trial (NASCET) entry criteria. The patients' mean age was 73 +/- 8 years (95 confidence interval [CI] 57 to 89), and 42 patients (25%) were > or = 80 years old. Patients had an independent neurologic examination before and after the procedure. RESULTS: The procedural success rate was 99%, including 73 patients who had a coronary intervention. Mean carotid artery stenosis was 78 +/- 10% before (95 CI 58 to 98) and 2 +/- 3% after the procedure (95 CI -4 to 8). During the initial hospital period and 30 days after CAS, there was one major and two category 2 minor strokes, as well as two category 1 minor strokes (total 30-day stroke rate was 2.9% for treated patients or 2.6% for treated arteries). There were no myocardial infarctions or deaths during or within 30 days of CAS. None of the NASCET-eligible patients had a stroke. At a mean follow-up of 19 +/- 11 months, three patients (2%) had asymptomatic restenosis. No other major strokes or neurologic deaths occurred. CONCLUSIONS: Carotid artery stenting is feasible, can be performed even in high risk patients and is associated with a low restenosis rate. PMID- 10841218 TI - A comparison of systematic stenting and conventional balloon angioplasty during primary percutaneous transluminal coronary angioplasty for acute myocardial infarction. STENTIM-2 Investigators. AB - OBJECTIVES: In a multicenter, randomized trial, systematic stenting using the Wiktor stent was compared to conventional balloon angioplasty with provisional stenting for the treatment of acute myocardial infarction (AMI). BACKGROUND: Primary angioplasty in AMI is limited by in-hospital recurrent ischemia and a high restenosis rate. METHODS: A total of 211 patients with AMI <12 h from symptom onset, with an occluded native coronary artery, were randomly assigned to systematic stenting (n = 101) or balloon angioplasty (n = 110). The primary end point was the binary six-month restenosis rate determined by core laboratory quantitative angiographic analysis. RESULTS: Angiographic success (Thrombolysis in Myocardial Infarction [TIMI] flow grade 3 and residual diameter stenosis <50%) was achieved in 86% of the patients in the stent group and in 82.7% of those in the balloon angioplasty group (p = 0.5). Compared with the 3% cross-over in the stent group, cross-over to stenting was required in 36.4% of patients in the balloon angioplasty group (p = 0.0001). Six-month binary restenosis (> or = 50% residual stenosis) rates were 25.3% in the stent group and 39.6% in the balloon angioplasty group (p = 0.04). At six months, the event-free survival rates were 81.2% in the stent group and 72.7% in the balloon angioplasty group (p = 0.14), and the repeat revascularization rates were 16.8% and 26.4%, respectively (p = 0.1). At one year, the event-free survival rates were 80.2% in the stent group and 71.8% in the balloon angioplasty group (p = 0.16), and the repeat revascularization rates were 17.8% and 28.2%, respectively (p = 0.1). CONCLUSIONS: In the setting of primary angioplasty for AMI, as compared with a strategy of conventional balloon angioplasty, systematic stenting using the Wiktor stent results in lower rates of angiographic restenosis. PMID- 10841220 TI - Acute hemodynamic and neurohumoral effects of selective ET(A) receptor blockade in patients with congestive heart failure. ET 003 Investigators. AB - OBJECTIVES: To investigate the hemodynamic effects of the selective endothelin (ET)A receptor antagonist LU135252 in patients with congestive heart failure (CHF). BACKGROUND: Nonselective ET(A/B( receptor antagonists improve hemodynamics in patients with CHF. Since ET(B( receptors mediate the release of nitric oxide and the clearance of ET-1, selective ET(A) antagonists are of special interest. METHODS: The hemodynamic effects of a single oral dose of the selective ET(A) receptor antagonist LU135252 (1, 10, 30, 100 or 300 mg) were investigated in a multicenter study involving 95 patients with CHF (New York Heart Association II III) with an ejection fraction < or = 35%. RESULTS: Baseline ET-1 positively correlated with pulmonary vascular resistance, pulmonary capillary wedge pressure (PCWP), and mean pulmonary artery pressure (MPAP, r = 0.37-0.50, p < 0.0004) but were inversely related to cardiac index (CI; r = -0.36, p = 0.0004). LU135252 dose dependently increased CI and decreased mean arterial pressure and systemic vascular resistance (p < 0.03-0.0002), while heart rate remained constant or decreased slightly. Pulmonary capillary wedge pressure, MPAP, pulmonary vascular resistance and right atrial pressure also decreased significantly (p < 0.035- < 0.0001). Two hours after LU135252, plasma ET-1 did not significantly increase after 1 mg but did so by 23% (p = 0.003), 29% (p = 0.0018), 56% (p < 0.0001) and 101% (p < 0.0001) after 10, 30, 100 and 300 mg, respectively, while plasma catecholamines remained constant. CONCLUSIONS: In patients with CHF, a single oral dose of the selective ET(A) receptor antagonist LU135252 improves hemodynamics in a dose-dependent manner without activation of other neurohumoral systems and is well tolerated over a wide dose range. PMID- 10841219 TI - The use of subcutaneous erythropoietin and intravenous iron for the treatment of the anemia of severe, resistant congestive heart failure improves cardiac and renal function and functional cardiac class, and markedly reduces hospitalizations. AB - OBJECTIVES: This study evaluated the prevalence and severity of anemia in patients with congestive heart failure (CHF) and the effect of its correction on cardiac and renal function and hospitalization. BACKGROUND: The prevalence and significance of mild anemia in patients with CHF is uncertain, and the role of erythropoietin with intravenous iron supplementation in treating this anemia is unknown. METHODS: In a retrospective study, the records of the 142 patients in our CHF clinic were reviewed to find the prevalence and severity of anemia (hemoglobin [Hb] <12 g). In an intervention study, 26 of these patients, despite maximally tolerated therapy of CHF for at least six months, still had had severe CHF and were also anemic. They were treated with subcutaneous erythropoietin and intravenous iron sufficient to increase the Hb to 12 g%. The doses of the CHF medications, except for diuretics, were not changed during the intervention period. RESULTS: The prevalence of anemia in the 142 patients increased with the severity of CHF, reaching 79.1% in those with New York Heart Association class IV. In the intervention study, the anemia of the 26 patients was treated for a mean of 7.2 +/- 5.5 months. The mean Hb level and mean left ventricular ejection fraction increased significantly. The mean number of hospitalizations fell by 91.9% compared with a similar period before the study. The New York Heart Association class fell significantly, as did the doses of oral and intravenous furosemide. The rate of fall of the glomerular filtration rate slowed with the treatment. CONCLUSIONS: Anemia is very common in CHF and its successful treatment is associated with a significant improvement in cardiac function, functional class, renal function and in a marked fall in the need for diuretics and hospitalization. PMID- 10841221 TI - Light-to-moderate alcohol consumption and prognosis in patients with left ventricular systolic dysfunction. AB - OBJECTIVES: The study evaluated the relationship between light-to-moderate alcohol consumption and prognosis in patients with left ventricular (LV) systolic dysfunction. BACKGROUND: Although chronic consumption of large amounts of alcohol can lead to cardiomyopathy, the effects of light-to-moderate alcohol consumption in patients with LV dysfunction are unknown. METHODS: The relationship between light-to-moderate alcohol consumption and prognosis was assessed in participants in the Studies of Left Ventricular Dysfunction (SOLVD), all of whom had ejection fraction values < or = 0.35. Baseline characteristics and event rates of patients who consumed 1 to 14 drinks per week (light-to-moderate drinkers, n = 2,594) were compared with those of patients who reported no alcohol consumption (nondrinkers, n = 3,719). The association between light-to-moderate alcohol consumption and prognosis was evaluated using Cox proportional hazards analysis, controlling for baseline differences and important covariates. RESULTS: Mortality rates were lower among light-to-moderate drinkers than among nondrinkers (7.2 vs. 9.4 deaths/100 person-years, p < 0.001). Among patients with ischemic LV dysfunction, light-to-moderate alcohol consumption was independently associated with a reduced risk of all-cause mortality (RR [relative risk] 0.85, p = 0.01), particularly for death from myocardial infarction (RR 0.55, p < 0.001). The risks of cardiovascular death, death from progressive heart failure, arrhythmic death, and hospitalization for heart failure were similar for light-to-moderate drinkers and nondrinkers in this group. Among patients with nonischemic LV dysfunction, light to-moderate alcohol consumption had no significant effect on mortality (RR 0.93, p = 0.5). CONCLUSIONS: Light-to-moderate alcohol consumption is not associated with an adverse prognosis in patients with LV systolic dysfunction, and it may reduce the risk of fatal myocardial infarction in patients with ischemic LV dysfunction. PMID- 10841222 TI - Prevalence and characteristics of dystrophin defects in adult male patients with dilated cardiomyopathy. AB - OBJECTIVES: To assess the prevalence of dystrophin defects in dilated cardiomyopathy (DCM) in male patients and to formulate investigation strategies for their identification. BACKGROUND: Dystrophin defects presenting with predominant or exclusive cardiac involvement may be clinically indistinguishable from "idiopathic" DCM. Diagnosis may be missed, unless specifically investigated. METHODS: Clinical and biochemical evaluation, right ventricular endomyocardial biopsy (EMB), light and electron microscopic and immunohistochemical studies of biopsy samples, six multiplex and two single polymerase chain reactions for 38 exons and automated sequencing of exon 9 and muscle promoter-exon 1 were undertaken in 201 consecutive male patients presenting with DCM, with (n = 14) and without (n = 187) increased serum creatine phosphokinase (sCPK). RESULTS: Dystrophin defects were identified in 13 of the 201 patients (6.5%, age 16-50). Family history was positive in four patients. Serum CPK levels were increased in 11 of 13 patients. Light microscopy examination of EMB was uninformative; ultrastructural study showed multiple membrane defects. Dystrophin immunostain was abnormal. Eight patients, all older than 20, had deletions affecting midrod domain, normal or mildly increased CPK and better outcome than the five remaining cases all younger than 20, with more than five-fold increase of sCPK. Two of these latter had proximal and rod-domain deletions. Sisters of two patients were diagnosed as noncarriers with microsatellite analysis. CONCLUSIONS: Although the overall prevalence of dystrophin defects in our consecutive DCM male series is low (6.5%), immunohistochemical and molecular studies are essential to identify protein and gene defects; screening studies are justified to define prevalence, clinical profile and genotype-phenotype correlation. PMID- 10841223 TI - Upregulation of the Bcl-2 family of proteins in end stage heart failure. AB - OBJECTIVES: To elucidate the pattern of expression of four members of the Bcl-2 family of proteins and to correlate this with terminal deoxynucleotidyl transferase [TdT]-mediated dUTP nick end labelling (TUNEL) and DNA fragmentation. BACKGROUND: Apoptosis has been implicated as a possible mechanism in the development of heart failure. However, the mechanisms involved remain unclear. METHODS: We have studied the expression of four members of the Bcl-2 family that are involved in the regulation of apoptosis and analyzed DNA fragmentation as a marker of apoptosis and as a biochemical criterion to distinguish between apoptosis and necrosis in dilated cardiomyopathy (DCM), ischemic heart disease (IHD) and normal donors. RESULTS: Western blot analysis and immunocytochemistry of the proapoptotic and antiapoptotic Bcl-2 proteins demonstrated significantly higher levels of all these proteins in the diseased groups compared with normal donors. Additionally, Bax was significantly higher in the IHD group compared with DCM. Terminal deoxynucleotidyl transferase [TdT]-mediated dUTP nick end labelling analysis demonstrated a significantly higher percentage of TUNEL-positive cells in the diseased groups compared with the control. Genomic DNA extraction of ventricular myocardial tissue showed no demonstrable DNA laddering for any of the groups. CONCLUSIONS: The significant increases in the levels of the proapoptotic proteins Bak and Bax and the higher percentage of TUNEL-positive cells in both diseased groups suggests the presence of ongoing apoptosis. However, increases in the antiapoptotic proteins, Bcl-2 and Bcl-xL, suggest a possible concomitant, compensatory antiapoptotic mechanism in patients with heart failure. PMID- 10841224 TI - The shift in the myocardial adenine nucleotide translocator isoform expression pattern is associated with an enteroviral infection in the absence of an active T cell dependent immune response in human inflammatory heart disease. AB - OBJECTIVES: This study evaluates the relevance of an enteroviral infection and the intramyocardial T-cell immune response for the alteration in the adenine nucleotide translocator isoform transcription pattern (ANTitp) in patients suspected of having myocardial inflammation. BACKGROUND: The ANT, the only mitochondrial carrier for ADP and ATP, plays a significant role in the energy metabolism and is involved in the apoptosis process. Its function and expression were found to be altered in the myocardium of patients with dilated cardiomyopathy and myocarditis. METHODS: The ANTitp was analyzed in endomyocardial biopsies from 53 patients with clinically suspected inflammatory heart disease (csIHD). Enteroviral RNA was detected in the biopsies using the reverse transcripted polymerase chain reaction technique. The activation of the cellular immune system was assessed by the quantification of T-lymphocytes employing immunohistochemistry. RESULTS: The ANTitp was found to be altered in 21 csIHD patients. Enteroviral genome was found in the heart of 71.4% of these patients, but only 37.5% of the patients with a normal ANTitp were virus-positive (p < 0.02). The infiltration with CD3+, CD45R0+ and CD8+ T-cells was substantially lower in myocardial specimens with an altered ANTitp than in biopsies with a normal ANTitp. Combining the data, an altered ANTitp was primarily found in virus-positive heart tissue, which was less infiltrated with lymphocytes or not at all. CONCLUSIONS: An enteroviral infection is linked to changes in the ANT isoform expression in human heart tissue, which shows little or no evidence of an active T-cell dependent immune response. These results make a contribution to a better understanding of the pathophysiology of enterovirus induced human inflammatory heart disease. PMID- 10841225 TI - Pericardial fluid from patients with unstable angina induces vascular endothelial cell apoptosis. AB - OBJECTIVES: The purpose of this study was to investigate whether pericardial fluid from patients with unstable angina (UA) would modulate vascular endothelial cell survival. BACKGROUND: Apoptosis of vascular endothelial cells promotes the coagulation process, playing an important role in the formation of coronary arterial thrombi. However, little is known about the mechanisms of vascular endothelial cell death in acute coronary syndrome. We hypothesized that factors inducing apoptosis are produced by the ischemic heart and accumulated in high concentrations in pericardial fluid. METHOD: Pericardial fluid was obtained during coronary artery bypass surgery from patients with UA (group A, n = 8) and those with stable angina (group B, n = 23). A survival assay of F2 cells from a mouse vascular endothelial cell line was performed in the presence of 10% pericardial fluid from each patient. RESULTS: Pericardial fluid levels of vascular endothelial growth factor were significantly higher in group A than in group B, indicating that group A had more ischemic insults than group B. Pericardial fluid from group A, but not from group B, markedly induced F2 cell death (cell survival relative to fetal bovine serum; group A: 33 +/- 26% vs. group B: 91 +/- 22%, p < 0.01). Cell death was associated with internucleosomal DNA fragmentation, a hallmark of apoptosis. Fractionation of pericardial fluid using a Centricon C-100 demonstrated that apoptosis-inducible activities exist in the Centricon C-100 retentates but not in the filtrates. CONCLUSIONS: Factors that induce vascular endothelial cell apoptosis are secreted into the pericardial space from the hearts of patients with UA. These factors are large complexes or unknown new proteins larger than 100 kDa. PMID- 10841226 TI - Risk stratification in unstable coronary artery disease--exercise test and troponin T from a gender perspective. FRISC-Study Group. Fragmin during InStability in Coronary artery disease. AB - OBJECTIVES: The study was done to determine the prognostic yield of an early symptom-limited exercise test (ET) and measurement of troponin T (TnT) in men and women with unstable coronary artery disease (CAD), with special reference to gender differences. BACKGROUND: Early risk assessment is essential for the application of appropriate treatment and further management in patients with unstable CAD. The early symptom-limited ET together with specific biochemical marker determination is an inexpensive, widely applicable method for early risk stratification. In women, however, the ET is considered less reliable, and there are few data on biochemical markers for risk stratification in women. METHODS: In a substudy of the Fragmin during InStability in Coronary artery disease (FRISC I) trial, 395 women and 778 men with unstable CAD who performed an early ET were followed for six months. Blood samples for TnT determination were taken in 342 women and 621 men at inclusion. RESULTS: Based on the ET results, low-, intermediate-, and high-risk response groups were identified with event rates of cardiac death or myocardial infarction (MI) of 1%, 9%, and 19%, respectively, among women and 8%, 14%, and 20%, respectively, among men. Patients who could not perform the ET had an event rate similar to the high-risk group. The TnT levels were divided into three groups: <0.06, 0.06-0.19, and > or = 0.20 microg/liter with event rates of 1%, 10%, and 18%, respectively, among women and 9%, 14%, and 18%, respectively, among men. Combining the ET results with TnT levels identified a low-risk group with an event rate of 3% in the male population and no events in the female population. CONCLUSIONS: Direct comparison between men and women from the same population with a high pretest likelihood of disease suggests that both TnT and the early symptom-limited ET are at least as useful as prognostic risk indicators in women as they are in men. PMID- 10841228 TI - The aspirin-angiotensin-converting enzyme inhibitor tradeoff: to halve and halve not. PMID- 10841227 TI - Clinical effects of early angiotensin-converting enzyme inhibitor treatment for acute myocardial infarction are similar in the presence and absence of aspirin: systematic overview of individual data from 96,712 randomized patients. Angiotensin-converting Enzyme Inhibitor Myocardial Infarction Collaborative Group. AB - OBJECTIVES: We sought to determine whether the clinical effects of early angiotensin-converting enzyme (ACE) inhibitor (ACEi) treatment for acute myocardial infarction (MI) are influenced by the concomitant use of aspirin (ASA). BACKGROUND: Aspirin and ACEi both reduce mortality when given early after MI. Aspirin inhibits the synthesis of vasodilating prostaglandins, and, in principle, this inhibition might antagonize some of the effects of ACEi. But it is uncertain whether, in practice, this influences the effects of ACEi on mortality and major morbidity after MI. METHODS: This overview sought individual patient data from all trials involving more than 1,000 patients randomly allocated to receive ACEi or control starting in the acute phase of MI (0-36 h from onset) and continuing for four to six weeks. Data on concomitant ASA use were available for 96,712 of 98,496 patients in four eligible trials (and for none of 1,556 patients in the one other eligible trial). RESULTS: Overall 30-day mortality was 7.1% among patients allocated to ACEi and 7.6% among those allocated to control, corresponding to a 7% (standard deviation [SD], 2%) proportional reduction (95% confidence interval 2% to 11%, p = 0.004). Angiotensin-converting enzyme inhibitor was associated with similar proportional reductions in 30-day mortality among the 86,484 patients who were taking ASA (6% [SD, 3%] reduction) and among the 10,228 patients who were not (10% [SD, 5%] reduction: chi-squared test of heterogeneity between these reductions = 0.4; p = 0.5). Angiotensin-converting enzyme inhibitor produced definite increases in the incidence of persistent hypotension (17.9% ACEi vs. 9.4% control) and of renal dysfunction (1.3% ACEi vs. 0.6% control), but there was no good evidence that these effects were different in the presence or absence of ASA (chi-squared for heterogeneity = 0.4 and 0.0, respectively; both not significant). Nor was there good evidence that the effects of ACEi on other clinical outcomes were changed by concomitant ASA use. CONCLUSIONS: Both ASA and ACEi are beneficial in acute MI. The present results support the early use of ACEi in acute MI, irrespective of whether or not ASA is being given. PMID- 10841229 TI - Prognostic significance of ST segment depression in lateral leads I, aVL, V5 and V6 on the admission electrocardiogram in patients with a first acute myocardial infarction without ST segment elevation. AB - OBJECTIVES: We sought to investigate the short-term prognostic value of the admission electrocardiogram (ECG) in patients with a first acute myocardial infarction (MI) without ST segment elevation. BACKGROUND: ST segment depression on hospital admission predicts a worse outcome in patients with a first acute MI, but the prognostic information provided by the location of ST segment depression remains unclear. METHODS: In 432 patients with a first acute MI without Q waves or > or = 0.1 mV of ST segment elevation, we evaluated the ability of the initial ECG to predict in-hospital death. RESULTS: The presence, magnitude and extent of ST segment depression were associated with an increased mortality, but the only electrocardiographic variable that was significant in predicting death after adjusting for baseline predictors was ST segment depression in two or more lateral (I, aVL, V5, or V6) leads (odds ratio 3.5, 95% confidence interval 1.2 to 10.6). Patients with lateral ST segment depression (n = 91, 21%) had higher rates of death (14.3% vs. 2.6%, p < 0.001), severe heart failure (14.3% vs. 4.1%, p < 0.001) and angina with electrocardiographic changes (20.0% vs. 11.6%, p = 0.04) than did the remaining patients, even though they had similar peak creatine kinase, MB fraction levels (129 +/- 96 vs. 122 +/- 92 IU/liter, p = NS). In contrast, ST segment depression not involving the lateral leads did not predict a poor outcome. Among patients who were catheterized, those with lateral ST segment depression had a lower left ventricular ejection fraction (57 +/- 12% vs. 66 +/- 13%, p = 0.001) and more frequent left main coronary artery or three-vessel disease than did the remaining patients (60% vs. 22%, p < 0.001). CONCLUSIONS: In patients with a first non-ST segment elevation acute MI, ST segment depression in the lateral leads on hospital admission predicts a poor in-hospital outcome. PMID- 10841230 TI - Acute myocardial infarction occurring in versus out of the hospital: patient characteristics and clinical outcome. Maximal Individual TheRapy in Acute Myocardial Infarction (MITRA) Study Group. AB - OBJECTIVES: We describe the baseline characteristics and clinical course of patients who had an acute myocardial infarction (AMI) during their hospital stay. BACKGROUND: In comparison with patients who had an AMI outside of the hospital (prehospital AMI), the data on patients who had an AMI in the hospital are poorly described. METHODS: Patients with an in-hospital AMI were prospectively registered in the Southwest German Maximal Individual TheRapy in Acute myocardial infarction (MITRA) study and compared with patients with prehospital AMI. RESULTS: Of 5,888 patients with AMI, 403 patients (6.8%) had an in-hospital AMI. These patients were older, more often male and sicker as compared with the patients with a prehospital AMI. They also showed a higher prevalence of concomitant diseases, such as arterial hypertension, diabetes mellitus, renal insufficiency and contraindications for thrombolysis. There was no significant difference regarding the use of reperfusion therapy, either thrombolysis (in hospital AMI 44.2% vs. prehospital AMI 49.1%; odds ratio [OR] 0.86, 95% confidence interval [CI] 0.70 to 1.05) or primary angioplasty (9.9% vs. 8.2%; OR 1.23, 95% CI 0.88 to 1.73), or a combination of both, between the two groups. The interval from symptom onset to the start of treatment in patients receiving reperfusion therapy was 55 min for patients with an in-hospital AMI versus 180 min for patients with a prehospital AMI (p = 0.001). In-hospital death occurred in 110 (27.3%) of 403 patients with an in-hospital versus 762 (13.9%) of 5,485 patients with a prehospital AMI (OR 2.33, 95% CI 1.85 to 2.94). This was confirmed by logistic regression analysis after adjusting for other confounding variables (OR 1.67, 95% CI 1.23 to 2.24). CONCLUSIONS: In-hospital AMI occurred in 6.8% of patients. Time to intervention was shorter; however, the use of reperfusion therapy for in-hospital AMI was not different from that for prehospital AMI. In particular, primary angioplasty seems to be underused in these patients. This, as well as the selection of patients, may result in the high hospital mortality rate of 27.3%. PMID- 10841231 TI - Cardiac troponin T in chest pain unit patients without ischemic electrocardiographic changes: angiographic correlates and long-term clinical outcomes. AB - OBJECTIVES: We prospectively evaluated the relation between cardiac troponin T (cTnT) level, the presence and severity of coronary artery disease (CAD) and long term prognosis in patients with chest pain but no ischemic electrocardiographic (ECG) changes who had short-term observation. BACKGROUND: Cardiac TnT is a powerful predictor of future myocardial infarction (MI) and death in patients with ECG evidence of an acute coronary syndrome. However, for patients with chest pain with normal ECGs, it has not been determined whether cTnT elevation is predictive of CAD and a poor long-term prognosis. METHODS: In 414 consecutive patients with no ischemic ECG changes who were triaged to a chest pain unit, cTnT and creatine kinase, MB fraction (CK-MB) were evaluated > or = 10 h after symptom onset. Patients with adverse cardiac events, including death, MI, unstable angina and heart failure were followed for as long as one year. RESULTS: A positive (>0.1 ng/ml) cTnT test was detected in 37 patients (8.9%). Coronary artery disease was found in 90% of 30 cTnT-positive patients versus 23% of 144 cTnT negative patients who underwent angiography (p < 0.001), with multivessel disease in 63% versus 13% (p < 0.001). The cTnT-positive patients had a significantly (p < 0.05) higher percent diameter stenosis and a greater frequency of calcified, complex and occlusive lesions. Follow-up was available in 405 patients (98%). By one year, 59 patients (14.6%) had adverse cardiac events. The cumulative adverse event rate was 32.4% in cTnT-positive patients versus 12.8% in cTnT-negative patients (p = 0.001). After adjustment for baseline clinical characteristics, positive cTnT was a stronger predictor of events (chi-square = 23.56, p = 0.0003) than positive CK-MB (>5 ng/ml) (chi-square = 21.08, p = 0.0008). In a model including both biochemical markers, CK-MB added no predictive information as compared with cTnT alone (chi-square = 23.57, p = 0.0006). CONCLUSIONS: In a group of patients with chest pain anticipated to have a low prevalence of CAD and a good prognosis, cTnT identifies a subgroup with a high prevalence of extensive and complex CAD and increased risk for long-term adverse outcomes. PMID- 10841232 TI - Coronary flow velocity immediately after primary coronary stenting as a predictor of ventricular wall motion recovery in acute myocardial infarction. AB - OBJECTIVES: The purpose of this study was to examine the relationship between the pattern of coronary blood flow velocity immediately after successful primary stenting and the recovery of left ventricular (LV) wall motion in patients with acute myocardial infarction (AMI). BACKGROUND: It is difficult to predict the recovery of LV wall motion immediately after direct angioplasty in AMI. Recent reports indicate that dysfunctional coronary microcirculation is an important determinant of prognosis for AMI patients after successful reperfusion. METHODS: We measured left anterior descending coronary flow velocity variables using a Doppler guide wire immediately after successful primary stenting in 31 patients with their first anterior AMI. The patients were divided into two groups: those with and those without early systolic reverse flow (ESRF). Changes in LV regional wall motion (RWM) and ejection fraction (EF) at admission and at discharge were compared between the two groups. Coronary flow velocity variables immediately after primary stenting were compared with changes in left ventriculographic indexes. RESULTS: The change in RWM was significantly greater in the non-ESRF group than it was in the ESRF group (0.9 +/- 0.7 vs. -0.1 +/- 0.3 standard deviation/chord, respectively, p < 0.001). The change in EF was also significantly greater in the non-ESRF group than it was in the ESRF group (10 +/- 10 vs. 1 +/- 6%, respectively, p < 0.05). In the non-ESRF group (diastolic to systolic velocity ratio [DSVR] <3.0), the DSVR correlated positively with the change in RWM (r = 0.60, p < 0.005, n = 24) and the change in EF (r = 0.52, p < 0.01). CONCLUSIONS: The coronary flow velocity pattern measured immediately after successful primary stenting is predictive of the recovery of regional and global LV function in patients with AMI. PMID- 10841233 TI - Measurement of postsystolic shortening to assess viability and predict recovery of left ventricular function after acute myocardial infarction. AB - OBJECTIVES: We sought to determine whether left ventricular (LV) postsystolic shortening in the region of acute myocardial infarction (MI) predicts functional recovery after primary angioplasty. BACKGROUND: Previous studies in experimental animals have shown that postsystolic shortening during temporary coronary occlusion predicts functional recovery after reperfusion. METHODS: Contrast ventriculography was performed on 35 patients with acute MI before and immediately after angioplasty, and one day, one month, three months and one year later. The centerline method was used to measure regional LV wall motion at end systole from all six ventriculograms as well as motion during isovolumic relaxation (motion(iso)) and postsystolic shortening from end systole until the end of contraction. The ventriculograms of 23 patients with normal anatomy were similarly analyzed. RESULTS: Wall motion at end systole improved significantly from baseline to follow-up in the infarct region. Postsystolic shortening at baseline correlated most closely with the recovery of wall motion at three months in patients with anterior infarction (r = 0.69, n = 25, p = 0.0001) but also with recovery at one month and one year. The correlation was slightly less powerful for motion(iso). Functional recovery could not be predicted from assessment of motion(iso) and postsystolic shortening in patients with inferior infarction. CONCLUSIONS: In patients with acute anterior MI, analysis of postsystolic shortening in the infarct region predicts the recovery of systolic LV function after reperfusion. Postsystolic shortening represents active contraction and indicates viable myocardium. PMID- 10841234 TI - Safety and clinical impact of ergonovine stress echocardiography for diagnosis of coronary vasospasm. AB - OBJECTIVES: We sought to address the issues of safety, feasibility and clinical impact of noninvasive diagnosis of coronary vasospasm (CVS). BACKGROUND: The safety of ergonovine provocation for CVS performed outside the catheterization laboratory has been questioned. METHODS: We performed a retrospective analysis of the results of bedside ergonovine provocation testing by monitoring left ventricular regional wall motion abnormalities (RWMAs) using two-dimensional echocardiography (Erg Echo). RESULTS: After confirming that there was no significant epicardial coronary stenosis, Erg Echo was performed on 1,372 patients from July 1991 to December 1997. Ergonovine echocardiography was terminated prematurely in 13 patients (0.9%) because of limitations caused by side effects unrelated to myocardial ischemia. Among 1,359 completed tests, 31% (n = 421) showed positive results, with development of RWMAs in 412 tests (98%) or ST displacement in electrocardiograms of nine tests (2%). Arrhythmias developed in 1.9% (26/1,372), including transient ventricular tachycardia (n = 2) and atrioventricular block (n = 4), which were promptly reversed with nitroglycerin. There was no mortality or development of myocardial infarction. Based on the angiographic criteria of 218 patients, the sensitivity and specificity of Erg Echo for the diagnosis of CVS were 93% and 91%, respectively. Since 1994, Erg Echo has become a more popular diagnostic method than invasive spasm provocation testing in the catheterization laboratory and has comprised more than 95% of all spasm provocation tests during the last three years. In the outpatient clinic, 453 patients underwent Erg Echo safely. CONCLUSIONS: Although this is a retrospective study in a single center, we believe that Erg Echo is highly feasible, accurate and safe for the diagnosis of CVS and can replace invasive angiographic spasm provocation testing in the catheterization laboratory. PMID- 10841235 TI - A renaissance of provocative testing for coronary spasm? PMID- 10841236 TI - Improvement of endothelial function and insulin sensitivity with vitamin C in patients with coronary spastic angina: possible role of reactive oxygen species. AB - OBJECTIVES: This study was designed to examine the effect of antioxidant supplementation on the endothelial function and insulin sensitivity in patients with coronary spastic angina (CSA). BACKGROUND: Insulin resistance may play a key role in coronary heart disease, and there is a possible link between acetylcholine-induced coronary vasoconstriction and hyperinsulinemia in patients with CSA. Endothelial dysfunction is present in the systemic arteries in CSA patients, and reactive oxygen species may cause inactivation of nitric oxide in these patients. METHODS: We measured flow-mediated dilation of the brachial artery using ultrasound technique in 22 patients with CSA and 20 control subjects. We also evaluated glucose tolerance using a 75-g oral glucose tolerance test and insulin sensitivity using steady-state plasma glucose (SSPG) methods in the same patients. RESULTS: The incidence of impaired glucose tolerance was higher in the CSA group than in the control group. Vitamin C infusion augmented flow-mediated dilation and decreased SSPG levels in the CSA group (from 3.27 +/- 0.77% to 7.00 +/- 0.59% [p < 0.001 by analysis of variance (ANOVA)] and from 177.3 +/- 13.3 to 143.1 +/- 14.9 mg/dl [p = 0.047 by ANOVA], respectively) but not in the control group (from 6.47 +/- 0.66% to 6.80 +/- 0.60% and from 119.8 +/ 11.7 mg/dl to 118.1 +/- 11.3 mg/dl, respectively). The steady-state plasma insulin levels were not affected by vitamin C infusion in either group. CONCLUSIONS: Vitamin C improves both endothelial function and insulin sensitivity in patients with CSA. Thus, reactive oxygen species and/or decreased nitric oxide bioactivity may play an important role in the genesis of both endothelial dysfunction and insulin resistance in patients with CSA. PMID- 10841237 TI - Myocardial perfusion and sympathetic innervation in patients with hypertrophic cardiomyopathy. AB - OBJECTIVES: This study assessed left ventricular myocardial perfusion and sympathetic innervation and function in hypertrophied and nonhypertrophied myocardial regions of patients with hypertrophic cardiomyopathy (HCM). BACKGROUND: Patients with HCM often have clinical findings consistent with increased cardiac sympathetic outflow. Little is known about the status of sympathetic innervation specifically in hypertrophic regions. METHODS: We conducted positron emission tomographic (PET) scanning using the perfusion imaging agent 13N-ammonia (13NH3) and the sympathoneuronal imaging agent 6-[18F] fluorodopamine (18F-FDA) in 8 patients with HCM and 15 normal volunteers. Positron emission tomographic data corrected for attenuation and the partial volume effect were analyzed using the region-of-interest technique. RESULTS: Myocardial 13NH3-derived radioactivity was similar in hypertrophied and nonhypertrophied regions of patients with HCM and in normal volunteers. At all time points, the 18F:13N ratio was lower in hypertrophied than in nonhypertrophied regions of HCM patients and in the septum of normal volunteers (p = 0.001). Trends in 18F-FDA-derived radioactivity over time were normal in both hypertrophied and nonhypertrophied myocardium. CONCLUSIONS: The results are consistent with decreased neuronal uptake of catecholamines in hypertrophied but not in nonhypertrophied myocardium of patients with HCM. Other aspects of cardiac sympathoneural function seem normal. Decreased neuronal uptake could reflect local relative hypoinnervation, decreased numbers of neuronal uptake sites, or metabolic limitations on cell membrane transport. By enhancing norepinephrine delivery to adrenoceptors for a given amount of sympathetic nerve traffic, decreased neuronal uptake can explain major clinical features of HCM. PMID- 10841238 TI - Thrombolysis is an effective and safe therapy in stuck bileaflet mitral valves in the absence of high-risk thrombi. AB - OBJECTIVES: We sought to evaluate the effectiveness and safety of thrombolytic therapy in stuck mitral bileaflet heart valves in the absence of high-risk thrombi. BACKGROUND: Current recommendations for the thrombolytic treatment of stuck prosthetic mitral valves are partially based on older valve models and inclusion of patients in whom high-risk thrombi were either ignored or not sought for. The feasibility and safety of thrombolysis in bileaflet models may be affected by the predilection of thrombi to catch the leaflet hinge. METHODS: We studied 12 consecutive patients (men/women = 5/7, age 58.8 +/- 14.9 years) who experienced one or more episodes of stuck bileaflet mitral valve over a 33-month period and received thrombolytic therapy with streptokinase, urokinase or tissue type plasminogen activator. Transesophageal echocardiography was performed in all patients. Patients with mobile or large (>5 mm) thrombi were excluded. Functional class at initial episode was I-II in 4 patients (33.3%) and III-IV in 8 patients (66.6%). RESULTS: Patients receiving thrombolytic therapy achieved an overall 83.3% freedom from a repeat operation or major complications (95% confidence interval 51.6-97.9%). Minor bleeding occurred in three patients (25%) and allergic reaction in one (8.3%). Transient vague neurologic complaints, without subjective findings, occurred in four patients (33.3%). Three patients had one or more relapses within 5.2 +/- 3.1 months from the previous episode, and readministration of thrombolytics was successful. CONCLUSIONS: In clinically stable patients with stuck bileaflet mitral valves and no high-risk thrombi, thrombolysis is highly successful and safe, both in the primary episode and in recurrence. The best thrombolytic regimen is yet to be established. PMID- 10841239 TI - Intravenous thrombolytic treatment of mechanical prosthetic valve thrombosis: a study using serial transesophageal echocardiography. AB - OBJECTIVE: We analyzed the results of intravenous thrombolytic treatment under transesophageal echocardiographic (TEE) guidance in prosthetic valve thrombosis. BACKGROUND: Thrombotic occlusion of prosthetic valves continues to be an uncommon but serious complication. Intravenous thrombolytic treatment has been proposed as an alternative to surgical intervention. METHODS: In a four-year period, 32 symptomatic patients with prosthetic valve related thrombosis underwent 54 thrombolytic treatment sessions for the treatment of 36 distinct episodes. All patients had low international normalized ratio values at the presentation. Transesophageal echocardiography was performed at baseline and repeated after each thrombolytic treatment session (total 98 TEE examinations). Streptokinase was used as the initial agent with a repeat dose given within 24 h when necessary. Recurrent thrombosis was treated either with tissue plasminogen activator or urokinase. RESULTS: The initial success after first dose was only 53% (17/32) but increased up to 88% (28/32) after repeated thrombolytic sessions upon documentation of suboptimal results on TEE examination (p < 0.01). In addition, four asymptomatic patients with large thrombi were also successfully treated with single infusion. The TEE characteristics of thrombus correlated with clinical presentation and response to lytics. Success was achieved with single lytic infusion in 40% of the obstructive thrombi as compared with 75% of the nonobstructive ones (p < 0.05). The success rates of lytic treatment were similar for mitral versus aortic valves, and for tilting disk versus bileaflet valves. Rapid (3 h) and slow (15 to 24 h) infusion of streptokinase resulted in similar success rates. However, major complications (three patients) occurred only in the rapid infusion group. CONCLUSION: In patients with prosthetic valve thrombosis, intravenous slow infusion thrombolysis given in discrete, successive sessions guided by serial TEE and transthoracic echocardiography can be achieved with a low risk of complications and a high rate of success. PMID- 10841240 TI - Clinical characteristics of a familial inherited myxomatous valvular dystrophy mapped to Xq28. AB - OBJECTIVES: The purpose of this study was to describe the phenotypic characteristics of an inherited myxomatous valvular dystrophy mapped to Xq28. BACKGROUND: Myxomatous valve dystrophies are a frequent cause of valvular diseases, the most common being idiopathic mitral valve prolapse. They form a group of heterogeneous diseases difficult to subclassify. The first mapping of the gene for a myxoid valvular dystrophy to Xq28 allowed investigation of the phenotype of affected members in a large family and characterization of the disease. METHODS: Among the 318 members in the pedigree, 89 agreed to participate in this study. Phenotypic characteristics were investigated using clinical examination, transthoracic echocardiography and biological analysis (F.VIII activity). Genetic status was based on haplotype analysis. RESULTS: Among 46 males, 9 were hemizygous to the mutant allele and had an obvious mitral and/or aortic myxomatous valve defect, and 4 had undergone valvular surgery. All had typical mitral valve prolapse associated in six cases with moderate to severe aortic regurgitation. The valve defect cosegregated with mild hemophilia A (F.VIII activity = 0.32 +/- 0.05). The 37 remaining males had normal valves and normal F.VIII activity. Heterozygous women were identified on the basis of their haplotypes. Among the 17 women heterozygous to the mutant allele, moderate mitral regurgitation was present in 8, associated with mild mitral valve prolapse in 1 and aortic regurgitation in 3, whereas 2 women had isolated mild aortic regurgitant murmur. In heterozygotes, the penetrance value was 0.60 but increased with age. CONCLUSION: X-linked myxomatous valvular disease is characterized by mitral valve dystrophy frequently associated with degeneration of the aortic valves affecting males and, to a lower severity, females. The first localization of a gene for myxomatous valvular diseases is the first step for the subclassification of these diseases. PMID- 10841241 TI - Prospective randomized comparison of antiarrhythmic therapy versus first-line radiofrequency ablation in patients with atrial flutter. AB - BACKGROUND: Despite the high success rate of radiofrequency (RF) ablation, pharmacologic therapy is still considered the standard initial therapeutic approach for atrial flutter. OBJECTIVE: We prospectively compared the outcome at follow-up of patients with atrial flutter randomly assigned to drug therapy or RF ablation. METHODS: Patients with at least two episodes of symptomatic atrial flutter in the last four months were randomized to regimens of either antiarrhythmic drug therapy or first-line RF ablation. After institution of therapy, end points included recurrence of atrial flutter, rehospitalization and quality of life. RESULTS: A total of 61 patients entered the study, 30 of whom were randomized to drug therapy and 31 to RF ablation. After a mean follow-up of 21 +/- 11 months, 11 of 30 (36%) patients receiving drugs were in sinus rhythm, versus 25 of 31 (80%) patients who underwent RF ablation (p < 0.01). Of the patients receiving drugs, 63% required one or more rehospitalizations, whereas post-RF ablation, only 22% of patients were rehospitalized (p < 0.01). Following RF ablation, 29% of patients developed atrial fibrillation which was seen in 53% of patients receiving medications (p < 0.05). Sense of well being (pre-RF 2.0 +/- 0.3 vs. post-RF 3.8 +/- 0.5, p < 0.01) and function in daily life (pre-RF 2.3 +/- 0.4 vs. post-RF 3.6 +/- 0.6, p < 0.01) improved after ablation, but did not change significantly in patients treated with drugs. CONCLUSION: In a selected group of patients with atrial flutter, RF ablation could be considered a first line therapy due to the better success rate and impact on quality of life, the lower occurrence of atrial fibrillation and the lower need for rehospitalization at follow-up. PMID- 10841242 TI - Catheter ablation of ventricular tachycardia in patients with structural heart disease using cooled radiofrequency energy: results of a prospective multicenter study. Cooled RF Multi Center Investigators Group. AB - OBJECTIVES: The purpose of this multicenter study was to evaluate the safety and efficacy of a radiofrequency (RF) catheter ablation system with internal saline irrigation. BACKGROUND: Catheter ablation of ventricular tachycardia (VT) associated with structural heart disease is more difficult than ablation of idiopathic VT. The larger size of responsible reentrant circuits contributes to the difficulty in achieving an adequate ablation lesion with conventional techniques. Recently, cooling of the ablation electrode by saline irrigation has been shown to increase RF lesion size. METHODS: The patient population included 146 patients who participated in the Cooled RF Ablation System clinical trial and underwent an attempt at ablation of VT occurring in the presence of structural heart disease. The duration of follow-up was 243 +/- 153 days. RESULTS: Catheter ablation was acutely successful, as defined by elimination of all mappable VTs, in 106 patients (75%). In 59 patients (41%), no VT of any type was inducible after ablation. Twelve patients (8%) experienced a major complication. After catheter ablation, 66 patients (46%) developed one or more episodes of a sustained ventricular arrhythmia. CONCLUSIONS: The results of this study demonstrate that catheter ablation of all mappable forms of sustained VT can be performed with high initial success and a moderate incidence of major complications (8%). PMID- 10841243 TI - A randomized trial comparing heparin initiation 6 h or 24 h after pacemaker or defibrillator implantation. AB - OBJECTIVES: The purpose of this randomized study was to evaluate the prevalence of pocket hematomas in patients treated with heparin 6 h or 24 h after pacemaker or defibrillator implantation. BACKGROUND: The risks of pocket hematoma and need for evacuation after device implantation have not been defined in patients who require anticoagulation. METHODS: Forty-nine consecutive patients with an indication for anticoagulation with heparin after implantable defibrillator or pacemaker implantation were randomized to receive intravenous heparin either 6 h (n = 26) or 24 h (n = 23) postoperatively. Both groups also received warfarin on a daily basis starting the evening of surgery. Twenty-eight patients who received postoperative warfarin alone and 115 patients who did not receive anticoagulation were followed up in a study registry. RESULTS: A pocket hematoma developed in 6 of 26 patients (22%) who were treated with intravenous heparin 6 h postoperatively, as compared with 4 of 23 patients (17%) who were treated with intravenous heparin 24 h postoperatively (p = 0.7). In total, a pocket hematoma developed in 10 of 49 patients (20%) treated with heparin, 1 of 28 patients (4%) treated with warfarin alone and 2 of 115 (2%) patients who received no anticoagulation (p < 0.001). CONCLUSIONS: Intravenous heparin initiation 6 h or 24 h after pacemaker or defibrillator implantation is associated with a 20% prevalence of pocket hematoma formation. Warfarin therapy or no anticoagulation is associated with only a 2% to 4% risk of pocket hematoma formation. PMID- 10841244 TI - Homozygosity for a HERG potassium channel mutation causes a severe form of long QT syndrome: identification of an apparent founder mutation in the Finns. AB - OBJECTIVES: We studied the clinical characteristics and molecular background underlying a severe phenotype of long QT syndrome (LQTS). BACKGROUND: Mutations of cardiac ion channel genes cause LQTS, manifesting as increased risk of ventricular tachycardia and sudden death. METHODS: We studied two siblings showing prolonged QT intervals corrected for heart rate (QTc), their asymptomatic parents with only marginally prolonged QTc intervals and their family members. The potassium channel gene HERG was screened for mutations by deoxyribonucleic acid sequencing, and the electrophysiologic consequences of the mutation were studied in vitro using the whole-cell patch-clamp technique. RESULTS: A novel missense mutation (L552S) in the HERG channel, present in the homozygous state in the affected siblings and in the heterozygous state in their parents, as well as in 38 additional subjects from six LQTS families, was identified. One of the homozygous siblings had 2:1 atrioventricular block immediately after birth, and died at the age of four years after experiencing unexplained hypoglycemia. The other sibling had an episode of torsade de pointes at the age of two years. The mean QTc interval differed significantly (p < 0.001) between heterozygous symptomatic mutation carriers (500 +/- 59 ms), asymptomatic mutation carriers (452 +/- 34 ms) and noncarriers (412 +/- 23 ms). When expressed in vitro, the HERG-L552S formed functional channels with increased activation and deactivation rates. CONCLUSIONS: Our data demonstrate that homozygosity for a HERG mutation can cause a severe cardiac repolarization disorder without other phenotypic abnormalities. Absence of functional HERG channels appears to be one cause for intrauterine and neonatal bradycardia and 2:1 atrioventricular block. PMID- 10841245 TI - A cross-sectional and diurnal study of thrombogenesis among patients with chronic atrial fibrillation. AB - OBJECTIVES: First, we sought to determine whether there is diurnal variation in hemostatic factors related to thrombogenesis and hypercoagulability among patients with chronic atrial fibrillation (AF). Second, we sought to determine whether levels of soluble thrombomodulin (sTM), a marker of endothelial function, or soluble P-selectin (sP-sel), an index of platelet activation, are altered in patients with AF as compared with subjects in sinus rhythm. BACKGROUND: Atrial fibrillation is associated with an increased risk of stroke and thromboembolism and is known to confer a hypercoagulable state, with abnormalities of thrombosis, platelet activation and endothelial cell function. Many cardiovascular events, such as acute myocardial infarction, have thrombosis as an underlying process, and they undergo diurnal variation. METHODS: Fifty-two patients (45 men, mean [+/ SD] age 66 +/- 6 years) with chronic AF, none of whom received antithrombotic therapy, were studied. Baseline levels of fibrinogen, sP-sel, sTM and von Willebrand factor (vWF) were compared to those levels in matched healthy control subjects in sinus rhythm. In a subgroup of 20 patients, five venous blood samples were collected through an indwelling cannula at 6-h intervals from 12 PM to 12 PM the following day and were analyzed for the same markers. RESULTS: Patients with chronic AF had higher plasma sP-sel, sTM, vWF and fibrinogen levels as compared with control subjects in sinus rhythm. Significant correlations were found between fibrinogen and sP-sel in patients with AF (r = 0.567 [Spearman], p < 0.001) and in control subjects (r = 0.334, p = 0.016). There was no significant diurnal variation in plasma levels of sP-sel, sTM, vWF or fibrinogen over the 24 h study period (repeated measures analysis of variance, p = NS). CONCLUSIONS: There is no circadian or diurnal variation in the hypercoagulable state seen in AF, as assessed by plasma fibrinogen and markers of platelet (sP-sel) and endothelial function (vWF and sTM). The persistent hypercoagulable state, together with the loss of diurnal variation in various hemostatic markers, in chronic AF may contribute to the high risk of stroke and thromboembolic complications in these patients. PMID- 10841246 TI - Clinical and echocardiographic characteristics of left atrial spontaneous echo contrast in sinus rhythm. AB - OBJECTIVES: In this study we attempt to define the clinical and echocardiographic characteristics of patients with left atrial spontaneous echo contrast (LASEC) in sinus rhythm (NSR). BACKGROUND: Left atrial spontaneous echo contrast in atrial fibrillation (AF) is associated with increased risk of thromboembolism. Little is known about its significance in NSR. METHODS: We reviewed reports of 1,288 transesophageal echocardiogram (TEE) studies done with a 5 MHz probe. Patients with swirling LASEC who were in NSR during TEE were analyzed. We compared them with a control group of 45 age matched patients selected to have NSR, left atrium (LA) > 4.0 cm but no SEC. RESULTS: Spontaneous echo contrast in NSR was noted in 24 patients (2%) and formed our study group. All patients with SEC had enlarged LA, mean 5.6 cm +/- 0.6 cm. There was a higher prevalence of cerebrovascular accident (CVA) in patients with SEC when compared with controls with no SEC, 83% versus 56%, p = 0.02. Patients with SEC had larger LA, 5.6 versus 4.9 cm, p < 0.0001 and lower mean peak left atrial appendage emptying velocity (LAAEV), 38 versus 56 cm/s, p = 0.001. Thirteen percent of patients with SEC had LA thrombus as compared with none in the control group, p = 0.02. By multivariate analysis, SEC in NSR was found to be associated with CVA, larger LA size and decreased mean LAAEV. Even after adjusting for LA size, patients with SEC had a higher prevalence of CVA than controls, p = 0.03. CONCLUSIONS: Spontaneous echo contrast in NSR occurs in patients with significantly dilated LA and depressed atrial function. Left atrial thrombus is noted in 13% of such patients despite NSR. Spontaneous echo contrast in NSR is associated with a higher prevalence of CVA. Further, SEC is found to be an independent and more powerful correlate of CVA than reduced LAAEV or atrial size. These data indicate that LASEC in NSR is a prothombotic condition. PMID- 10841247 TI - Multisite pacing for prevention of atrial tachyarrhythmias: potential mechanisms. AB - OBJECTIVES: To determine the effects of single-, dual-, triple- and quadruple site atrial pacing on atrial activation and refractoriness in normal canine hearts. BACKGROUND: Multisite pacing has been suggested to be superior to single site pacing for prevention of atrial tachyarrhythmias. However, the underlying electrophysiological mechanisms are undetermined at the moment, as is the rationale for the selection of pacing locations and the number of pacing sites. METHODS: In 13 normal beagle dogs, an epicardial multielectrode (128 bipoles) and a multiplexer mapping system were used to reconstruct epicardial atrial activation patterns obtained during simultaneous stimulation from up to four electrodes located in the high and low right and left atrium, respectively. For all pacing modes (single-, dual-, triple- and quadruple-site pacing), total activation times and local effective refractory periods at eight randomly selected sites as well as local recovery intervals were determined. In a subgroup of five dogs, total epicardial activation times were also obtained during single site septal stimulation (septal group). RESULTS: Activation times and local recovery intervals were minimized by triple-site stimulation, whereas a fourth site did not produce further shortening. Septal stimulation produced epicardial activation times comparable to quadruple-site stimulation. Local refractory periods and their dispersion always remained unaffected. Functional conduction blocks apparent during single-site were found to resolve during multisite stimulation. CONCLUSIONS: Multisite pacing can prevent functional conduction blocks by multidirectional excitation and a reduction in total activation time. Triple-site and, possibly, septal pacing modes are expected to be most efficient because both minimize total activation times and maximize the multidirectionality of excitation. In spite of unaffected local refractory periods, the shortening of local recovery intervals might homogenize atrial repolarization and, thus, contribute to the preventive effects of multisite pacing. PMID- 10841248 TI - Determinants of forward pulmonary vein flow: an open pericardium pig model. AB - OBJECTIVE: To elucidate determinants of pulmonary venous (PV) flow. BACKGROUND: Right ventricular (RV) systolic pressure (vis a tergo), left atrial (LA) relaxation and left ventricular (LV) systole and relaxation (vis a fronte) have been suggested as determinants of the pulmonary venous (PV) anterograde Doppler flow velocities, but their relative contributions to those flow velocities have not been quantified. METHODS: We analyzed, by multiple regression analysis, the determinants of PV anterograde velocities in an open-pericardium, paced (70 and 90 beats/min) pig model in which LA afterload was modified by creating LV regional ischemia (left anterior descending coronary artery constriction). We measured high fidelity LA, LV and RV pressures and Doppler flow velocities (epicardial echocardiography). We calculated LV tau, LA relaxation (a through x pressure difference divided by time, normalized by a pressure), LA peak v through x and RV systolic through LA peak v (RVSP-v) pressure differences, LV ejection fraction, long-axis shortening, stroke volume (LV outflow integral x outflow area) and LA four-chamber dimensions, Doppler transmitral and PV flow velocities and velocity-time integrals. RESULTS: Left ventricular regional ischemia increased mildly LA y trough pressure (8 +/- 1 vs. 6 +/- 1 mm Hg, p = 0.001). Left ventricular stroke volume (coefficient: 0.5 cm/ml, SE: 0.2, p = 0.005) and LA peak v pressure (coefficient: -0.8 cm/mm Hg, SE: 0.3, p = 0.008) determined the PV total systolic integral. Left atrial relaxation determined both PV early systolic peak velocity and integral (coefficient: -0.8 cm/mm Hg, SE: 0.3, p = 0.04). Left atrial maximum area (coefficient: 2 cm(-1) SE: 0.7, p = 0.01) and RVSP-v (coefficient: 0.1 cm/mm Hg, SE: 0.05, p = 0.03) determined the late systolic integral. The PV total systolic integral determined both PV early diastolic peak velocity and integral (coefficient: 1.2, SE: 0.2, p = 0.001). CONCLUSIONS: In an experimental model of LV acute ischemia of limited duration, the main independent predictors of PV systolic anterograde flow velocities are LA relaxation and compliance (LA peak v pressure) and LV systole--all vis a fronte factors. In the setting of mildly increased LA pressures, PV systolic flow (LA reservoir filling) is an independent predictor of PV early diastolic flow (LA early conduit). PMID- 10841249 TI - Response to incremental doses of dobutamine early after reperfusion is predictive of the degree of myocardial salvage in dogs with experimental acute myocardial infarction. AB - OBJECTIVES: We sought to determine whether the inotropic response to dobutamine might be useful for estimating the extent of viable myocardium soon after reperfusion. BACKGROUND: Early identification of viable myocardium in the presence of severe left ventricular dysfunction after reperfusion is important for clinical decision making. METHODS: Nine open-chest dogs had left anterior descending coronary artery occlusion for 40 to 180 min, followed by gradual reperfusion. The systolic thickening response to incremental dobutamine doses was measured with ultrasonic crystals and regional flow by microspheres. RESULTS: Dogs were divided into two groups based on triphenyl tetralozium chloride infarct size (group 1: 9.3 +/- 3.0% risk area; group 2: 51.1 +/- 4.8%). In group 2 dogs with larger infarcts, regional flow during peak dobutamine was lower than it was in group 1 in endocardial (1.15 +/- 0.22 vs. 2.64 +/- 0.33 mL x min(-1) x g(-1)) and midwall (1.47 +/- 0.32 vs. 2.92 +/- 0.36 mL x min(-1) x g(-1)) layers, and endocardial flow in group 2 failed to increase from baseline (0.96 +/- 0.07 vs. 1.15 +/- 0.22 mL x min(-1) x g(-1)). Group 1 dogs demonstrated a dose dependent increase in systolic thickening with dobutamine versus a blunted response in group 2. The inotropic response to only 10 microg x kg(-1) x min(-1) of dobutamine was predictive of the degree of myocardial salvage. CONCLUSIONS: In the early postischemic stunning phase of reperfusion, the inotropic response to dobutamine is predictive of the degree of myocardial salvage and ultimate infarct size. The ability to distinguish between stunned versus necrotic myocardium early after reperfusion was most likely due to the presence of subendocardial flow reserve during dobutamine in dogs with predominantly salvaged myocardium. PMID- 10841250 TI - Local paclitaxel delivery for the prevention of restenosis: biological effects and efficacy in vivo. AB - OBJECTIVE: The aim of this study was to evaluate the potential of paclitaxel to prevent restenosis in vivo. BACKGROUND: Paclitaxel (Taxol) is a microtubule stabilizing compound with potent antitumor activity. It influences the cytoskeleton equilibrium by increasing the assembly of altered microtubules, thereby inducing cellular modifications that result in reduced proliferation, migration and signal transduction. METHODS: Before the in vivo study, delivery efficiency was determined with radiolabeled paclitaxel in porcine hearts. After induction of a defined plaque in the right carotid arteries of 76 New Zealand rabbits by electrical stimulation, 27 animals underwent balloon dilation and subsequent local paclitaxel delivery (10 ml, 10 micromol/liter) with a double balloon catheter. Twenty-nine animals served as control with angioplasty only, 10 animals underwent local delivery of vehicle only (0.9% NaCl solution) and 10 animals were solely electrostimulated. Vessels were excised one, four, and eight weeks after intervention. RESULTS: The extent of stenosis in paclitaxel-treated animals was significantly reduced compared with balloon-dilated control animals (p = 0.0012, one, four and eight weeks after intervention: 14.6%, 24.6% and 20.5%, vs. 24.9%, 33.8% and 43.1%, respectively). Marked vessel enlargement compared with balloon-dilated control animals could be observed (p = 0.0001, total vessel area after one, four and eight weeks: paclitaxel group: 1.983, 1.700 and 1.602 mm2, control: 1.071, 1.338 and 1.206 mm2, respectively). Tubulin staining and electron microscopy revealed changes in microtubule assembly, which were limited to the intimal area. Vasocontractile function after paclitaxel treatment showed major impairment. CONCLUSIONS: Local delivery of paclitaxel resulted in reduced neointimal stenosis and enlargement in vessel size. Both these effects contribute to a preservation of vessel shape and are likely to be caused by a structural alteration of the cytoskeleton. PMID- 10841251 TI - Plant-derived estrogens relax coronary arteries in vitro by a calcium antagonistic mechanism. AB - OBJECTIVES: To investigate the potential for plant derived estrogens (phytoestrogens) genistein, phloretin, biochanin A and zearalanone to relax rabbit coronary arteries in vitro and to determine the mechanism(s) of such relaxation. BACKGROUND: Epidemiological data suggests a reduction in the incidence of coronary heart disease in humans who have a high intake of phytoestrogens. METHODS: Isolated rabbit coronary artery rings were suspended in individual organ baths, precontracted with potassium chloride (30 mM), and the relaxing effects and mechanisms of relaxation to genistein, phloretin, biochanin A and zearalanone were determined by measurement of isometric tension. RESULTS: Genistein, phloretin and biochanin A induced significant gender-independent relaxation in rings with and without endothelium. Inhibition of nitric oxide and prostaglandin synthesis with L-NAME and indomethacin had no effect on genistein induced relaxation. Relaxation was unaffected by the specific estrogen receptor antagonist ICI 182,780, the ATP-sensitive potassium channel inhibitor glibenclamide and the potassium channel inhibitor, barium chloride. Calcium concentration-dependent contraction curves in high potassium depolarization medium were significantly shifted to the right and downward after incubation with genistein and zearalanone. An inhibitory effect of genistein (2 microM) on L-type calcium current in guinea-pig ventricular myocytes confirmed a calcium antagonist relaxing mechanism of action. In healthy volunteers, plasma genistein levels of approximately 2 microM are achieved after ingestion of a commercially available soy protein drink (Supro) containing 37 mg genistein. CONCLUSIONS: This study demonstrates that phytoestrogens induce endothelium-independent relaxation of coronary arteries; the mechanism involves calcium antagonism. These mechanisms may contribute to the potential long-term cardiovascular protective effect of these substances. PMID- 10841253 TI - President's page: ACC takes strategic steps to address members' needs. American College of Cardiology. PMID- 10841252 TI - Phytoestrogens and vascular therapy. PMID- 10841254 TI - Beta-blockers and ventricular arrhythmias in dilated cardiomyopathy. PMID- 10841255 TI - Pharmacologic stress echocardiography: can we forget "state-of-the-art" protocols? PMID- 10841256 TI - Overdosing with prostacyclin in primary pulmonary hypertension. PMID- 10841257 TI - Intracellular pH regulation and buffer capacity in CO2/HCO3-buffered media in cultured epithelial cells from rainbow trout gills. AB - The influence of a CO2/HCO3(-)-buffered medium on intracellular pH regulation of gill pavement cells from freshwater rainbow trout was examined in monolayers grown in primary culture on glass coverslips; intracellular pH (pHi) was monitored by continuous spectrofluorometric recording from cells loaded with 2',7'-bis(2-carboxyethyl)-5(6)-carboxy-fluoroscein. When cells in HEPES-buffered medium at normal pH = 7.70 were transferred to normal CO2/HCO3(-)-buffered medium ?PCO2 = 3.71 mmHg, [HCO3-] = 6.1 mmol l(-1), extracellular pH (pHe) = 7.70?, they exhibited a brief acidosis but subsequently regulated the same pHi (approximately 7.41) as in HEPES. Buffer capacity (beta) increased by the expected amount (5.5 8.0 slykes) based on intracellular [HCO3-], and was unaffected by most drugs and treatments. However, after transfer to high PCO2 = 11.15 mmHg, [HCO3-] = 18.2 mmol l(-1) at the same pHe = 7.70, the final regulated pHi was elevated (approximately 7.53). The rate of correction of alkalosis caused by washout of this high PCO2, high-HCO3- medium was unaffected by removal of extracellular Cl-. Removal of extracellular Na + lowered resting pHi and greatly inhibited the rate of pHi recovery from acidosis. Bafilomycin A1 (3 micromol l(-1)) had no effect on these responses. However amiloride (0.2 mmol l(-1)) inhibited recovery from acidosis caused by washout of an ammonia prepulse, but did not affect resting pHi, the latter differing from the response in HEPES where amiloride also lowered resting pHi. Similarly 4-acetamido-4'- isothiocyanatostilbene-2,2'-disulfonic acid, sodium salt (0. 1 mmol l(-1)) did not affect resting pHi but slowed the rate of recovery from acidosis, though to a lesser extent than amiloride. Removal of extracellular Cl(-1) also slowed the rate of recovery but greatly increased beta by an unknown mechanism; when this was taken into account, H+ extrusion rate was unaffected. These results are consistent with the presence of Na+ -(HCO3)N co transport and/or Na+-dependent HCO3(-)/Cl(-) exchange, in addition to Na+/H+ exchange, as mechanisms contributing to "housekeeping" pHi regulation in gill cells in CO2/HCO3(-) media, whereas only Na+/H+ exchange is seen in HEPES. Both Na(+)-independent Cl-/HCO3(-) exchange and V-type H(+)-ATPase mechanisms appear to be absent from these cells cultured in isotonic media. PMID- 10841258 TI - Plant secondary metabolites as mammalian feeding deterrents: separating the effects of the taste of salicin from its post-ingestive consequences in the common brushtail possum (Trichosurus vulpecula). AB - The effect of the phenolic glycoside, salicin, on food intake of the common brushtail possum (Trichosurus vulpecula) was studied in a series of feeding experiments. Increasing the concentration of salicin in a diet of fruits and cereals led to significant reductions of food intake in the short term (6 days). After prolonged (20 days) exposure to salicin, food intake (19 g kg(-0.75) day( 1)) was still reduced relative to controls (31 g kg(-0.75) day(-1)) but not reduced to the same extent as in the short-term experiments. Nonetheless, over these 20 days, common brushtail possums regulated their intake of salicin so as not to exceed a threshold limit of 1.9 +/- 0.1 g kg(-0.75) day(-1). Manipulative experiments sought to determine whether this threshold intake was in response to pre-ingestive factors (taste) or the post-ingestive consequences of ingesting salicin. Dietary salicin (0.17-5.0% DM) had no significant effect on nitrogen balance or urea metabolism and injection of a specific serotonin receptor antagonist, ondansetron, did not lead to increases in salicin intake as has been found for some other plant secondary metabolites. Similarly, administration of 1.3 g salicin by gavage had no significant effect on the subsequent intake of salicin compared to controls that were gavaged with water. We concluded that pre ingestive factors were responsible for common brushtail possums limiting their intake of salicin-rich diets rather than any measurable post-ingestive consequence of feeding. PMID- 10841259 TI - Compliance of the respiratory system in newborn and adult rats after gestation in hypoxia. AB - We hypothesized that hypoxia during gestation modifies the compliance of the respiratory system of newborn and adult rats. Pregnant rats were placed in a hypobaric chamber at an inspired oxygen pressure of 86 mmHg (equivalent to 12% O2 in normobaria) from day 4 of gestation until day 2 post-partum. Three-day-old rat pups were smaller than controls, with higher hematocrit; the lungs were also small, with less protein and DNA content. The pressure (x-axis)-volume (y-axis) curve of the respiratory system was displaced to the right of the control curve, and the compliance of the respiratory system, measured on the inflation or deflation limb of the pressure-volume curve, was decreased by approximately 20 25%, depending upon the normalization procedure (per body mass or per dry lung weight). In 50-day-old rats exposed to hypoxia during gestation, body weight, hematocrit, lung mass and DNA content were normal; the compliance of the respiratory system, measured at ventilation frequencies between 20 cpm and 100 cpm, was higher than in controls by approximately 20%. It is concluded that the effects of prenatal hypoxia on the compliance of the respiratory system can vary with age. In the rat the process of alveolar formation initiates postnatally. Hence, in the newborn the effects of the prenatal hypoxia on the compliance of the respiratory system are likely to be dominated by the hypoxic pulmonary hypoplasia and hypertension, which decrease the compliance of the respiratory system. In the adult, the effects of the decreased alveolar formation are the prevailing ones, increasing the compliance of the respiratory system. PMID- 10841261 TI - How well can common brushtail possums regulate their intake of Eucalyptus toxins? AB - We studied factors affecting the ability of common brushtail possums (Trichosurus vulpecula) to regulate their intake of a dietary toxin, jensenone, extracted from Eucalyptus leaves. Increasing concentrations of jensenone in the diet led to a dose-dependent decrease in food intake best described as an exponential decay. Animals that had not previously been exposed to jensenone ate significantly more when first offered food containing the compound than on subsequent days. However, when offered the same amount of food in a number of portions throughout the night, naive animals ate significantly less than animals offered the total meal at once. When offered food containing jensenone over a 13-day period, the animals' intake varied cyclically with relatively high food intakes followed by relatively low intakes. Furthermore, animals that were exposed to cold conditions (4 degrees C) ate more than those maintained at 18 degrees C but this difference was abolished when jensenone was included in the diet. We interpret these results as showing that regulation of toxin intake by common brushtail possums depends on learned responses that can override other important influences on feeding. PMID- 10841260 TI - Dietary phosphorus regulates intestinal transport and plasma concentrations of phosphate in rainbow trout. AB - Intestinal inorganic phosphate transport and its regulation have not been studied in fish. In this study, we initially characterized the mechanisms of intestinal inorganic phosphate transport in rainbow trout (Oncorhynchus mykiss) then determined the effects of dietary phosphorus concentrations on intestinal inorganic phosphate uptake, plasma inorganic phosphate, and intestinal luminal inorganic phosphate concentrations. In 11-g trout, the saturable mechanism of brushborder inorganic phosphate uptake had a Kt= 1.2 mmol l(-1) and a Vmax = 0.22 nmol mg(-1) min(-1), while the diffusive component had a Kd = 0.012 min(-1). Similar kinetic constants were obtained from 51-g trout, suggesting that development or size had little effect on transport. Tracer inorganic phosphate (1.18 mmol l(-1)) uptake was almost completely inhibited (>95%) by 20 mmol l(-1) unlabeled inorganic phosphate. Inorganic phosphate uptake (0.2 mmol l(-1)) was strongly inhibited (approximately 75% inhibition) by phosphonoformic acid, a competitive inhibitor of mammalian inorganic phosphate transport, as well as by the absence of Na+ (approximately 90% inhibition). Northern blot and reverse transcription-polymerase chain reaction indicated that the intestinal inorganic phosphate transporter in trout is not related to the cloned Na+ inorganic phosphate-II transporter of winter flounder. Intestinal luminal and plasma inorganic phosphate concentrations each increased with dietary P concentrations. Intestinal inorganic phosphate, but not proline, absorption rates decreased with dietary phosphorus concentrations. As in mammals and birds, a Na-dependent inorganic phosphate carrier that is tightly regulated by diet is present in trout small intestine. PMID- 10841262 TI - A putative hyperglycemic factor from the cerebral ganglia of Otala lactea (Mollusca: Pulmonata). AB - Mantle tissue pieces from adult Otala lactea continuously synthesized glycogen over a 72-h incubation period. Acid-saline extract of the cerebral ganglia inhibited glycogen synthesis by mantle tissue in vitro. This effect was dose dependent. The glycogen reduction factor from the cerebral ganglia was heat stable, protease sensitive, and relatively hydrophobic. The cerebral ganglia extract also stimulated mantle glycogen phosphorylase in vitro in a dose dependent manner. The results suggest the presence of a hyperglycemic factor in the cerebral ganglia of Otala. The molecular weight of this factor, estimated by size-exclusion chromatography, was approximately 10,000. Mammalian glucagon had no significant effect on glycogen synthesis by the mantle pieces. PMID- 10841263 TI - Effects of strontium on the absorption of calcium, magnesium and phosphate ions from the ovine reticulo-rumen. AB - The net absorption rates of strontium ions from the ovine reticulo-rumen, isolated in situ in trained conscious animals, were measured under controlled conditions. A linear positive response was obtained from the addition of Sr2+ ions to the artificial rumen fluid. This increase in the absorption of Sr was reflected in an increase in the plasma Sr concentration. In contrast to the discrimination observed elsewhere in favour of the absorption of Ca relative to Sr, the absorption rate of Sr from the reticulo-rumen was significantly greater than that of Ca, from solutions containing the same molar concentration. A graded increase in the Sr concentration in the ruminal fluid from 1 mmol/l to 4 mmol/l led to a corresponding reduction in the absorption rate of Ca but an increase in that of phosphate. The latter result is similar to that observed when the intra ruminal concentration of Ca2+ ions is increased. It is suggested that Ca and Sr share a common pathway for absorption from the reticulo-rumen and that this may involve coupling with the absorption of phosphate ions. PMID- 10841264 TI - Effects of ambient PO2 and temperature on oxygen uptake in Nautilus pompilius. AB - This study employs closed-circuit respirometry to evaluate the effect of declining ambient oxygen partial pressure (PO2) and temperature on mass specific rates of oxygen uptake (VO2) in Nautilus pompilius. At all temperatures investigated (11, 16, and 21 degrees C), VO2 is relatively constant at high PO2 (oxyregulation) but declines sharply at low PO2 (oxyconformation). The critical PO2 below which oxyconformation begins (Pc) is temperature dependent, higher at 21 degrees C (49 mmHg) than at 11 degrees C or 16 degrees C (21.7 mmHg and 30.8 mmHg respectively). In resting, post-absorptive animals, steady-state resting VO2 increases significantly with temperature resulting in a Q10 value of approximately 2.5. The metabolic strategy of N. pompilius appears well suited to its lifestyle, providing sufficient metabolic scope for its extensive daily vertical migrations, but allowing for metabolic suppression when PO2 falls too low. The combination of low temperatures and low PO2 may suppress metabolic rate 16-fold (assuming negligible contributions from anaerobic metabolism and internal O2 stores), enhancing hypoxia tolerance. PMID- 10841265 TI - Uptake of 3-O-methyl-D-[U-14C]glucose into brain of rainbow trout: possible effects of melatonin. AB - The influx of glucose into the brain and plasma glucose disappearance were estimated in rainbow trout (Oncorhynchus mykiss) intravenously injected (1 ml x kg(-1) body weight) with a single dose (15 microCi x kg(-1) body weight) of 3-O methyl-D-[U-14C]glucose ([U-14C]-3-OMG) at different times (2-160 min), and after intravenous injection at 15 min of increased doses (10-60 microCi x kg(-1) body weight) of [U-14C]-3-OMG. Brain and plasma radiotracer concentrations were measured, and several kinetic parameters were calculated. The apparent brain glucose influx showed a maximum after 15-20 min of injection then decreased to a plateau after 80 min. Brain distribution space of 3-OMG increased from 2 min to 20 min reaching equilibrium from that time onwards at a value of 0.14 ml x g(-1). The unidirectional clearance of glucose from blood to brain (k1) and the fractional clearance of glucose from brain to blood (k2) were estimated to be 0.093 m x min(-1) x g(-1), and 0.867 min(-1), respectively. A linear increase was observed in brain and plasma radiotracer concentrations when increased doses of [U-(14)C]3-OMG were used. All these findings support a facilitative transport of glucose through the blood-brain barrier of rainbow trout with characteristics similar to those observed in mammals. The injection of different doses of melatonin (0.25-1.0 mg x kg(-1)) significantly increased brain glucose influx suggesting a possible role for melatonin in the regulation of glucose transport into the brain. PMID- 10841266 TI - Seasonal changes in plasma testosterone and glucocorticosteroids in free-living male yellow-pine chipmunks and the response to capture and handling. AB - We measured plasma levels of testosterone, corticosterone, and cortisol in free living male yellow-pine chipmunks to demonstrate the patterns of seasonal variation and to assess the effects of capture and handling on hormone levels. We achieved the latter by modifying our standard trapping technique (blood samples collected within 1-3 h of capture) to obtain blood samples that allowed measurement of hormone levels within 3 min of capture (basal) and again 30 min later. By alternating the modified and standard trapping techniques over 7 months of the active season we demonstrated that seasonal patterns of variation in steroid hormone levels can be accurately described with the simpler, standard trapping technique. Basal and 30-min post-capture testosterone levels were high during mating and dropped to a persistently low level thereafter. Conversely, both cortisol and corticosterone were at their seasonal low during mating and climbed to peak levels in June following reproduction. Plasma glucocorticosteroid levels increased during the 30 min after capture and handling at all times of the active season, and these elevated levels were similar to the levels obtained by standard trapping. Testosterone levels during the mating period also increased in response to capture and handling. The contrasting patterns of seasonal variation in glucocorticosteroid and testosterone levels and the changes induced by capture and handling suggest that when testosterone concentration is high, adrenocortical activity is suppressed. PMID- 10841267 TI - Body reserves and nutritional needs during laying preparation in barn owls. AB - To investigate the composition of the body reserves made during pre-laying by breeding European barn owls (Tyto alba), we have analysed the body composition of captive breeding and non-breeding females sacrificed during the laying period. The data obtained were compared to the daily requirement for egg formation obtained by an egg composition analysis and the timing of egg synthesis. This study demonstrates that body mass gain observed in breeding females (+ 38.3 g after eggs in formation and gonadal tractus were removed) was not the consequence of an accumulation of body fuels like lipids but of mainly water and lean material. The lipidic reserves were found to be less important in breeding than in non-breeding females and their localisation modified; lipids were absent from medullar bones in breeding females which liberated room for other storage. The subcutaneous tissue, which was homogeneous in non-breeding females, was located principally under the brood patch in breeding females. Nutrients and energy required during egg formation could be obtained without modification of daily food intake. These results show that a laying event can be initiated in 14 days and that the onset of reproduction is not triggered by body condition in barn owls. The water accumulation observed is suggested to be the mere consequence of an increase of protein metabolism (egg and moult). These results intimate that the body mass increase observed in diurnal and nocturnal raptors during laying preparation, interpreted as an energy safety strategy, ought to be reconsidered. PMID- 10841268 TI - The glucose concentration modulates N-formyl-methionyl-leucyl-phenylalanine (fMet Leu-Phe)-stimulated chemokinesis in normal human neutrophils. AB - The effects of glucose concentration on the chemokinetic effects of the chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine (fMet-Leu-Phe) was evaluated for normal human neutrophils using a direct microscopic assay. fMet-Leu Phe increased the rate of locomotion in the absence of glucose, but the chemokinetic effect of fMet-Leu-Phe was most potent at 5 mM glucose and not further changed at 15 mM glucose. The chemokinetic effects of fMet-Leu-Phe and glucose were essentially the same in blood clot-isolated and gradient-isolated neutrophils. However, in gradient-isolated neutrophils, the rate of locomotion under different experimental conditions was strictly negatively correlated to the fraction of non-locomoting cells and the degree of adhesion to the substratum. These results indicate that the chemokinetic effects of fMet-Leu-Phe are regulated by the glucose concentration by inducing locomotor activity in otherwise non-locomoting cells and by improving adhesion to the substratum. PMID- 10841269 TI - Inhibition of mitochondrial permeability transition by low pH is associated with less extensive membrane protein thiol oxidation. AB - Ca2+ and inorganic phosphate-induced mitochondrial swelling and membrane protein thiol oxidation, which are associated with mitochondrial permeability transition, are inhibited by progressively decreasing the incubation medium pH between 7.2 and 6.0. Nevertheless, the detection of mitochondrial H2O2 production under these conditions is increased. Permeability transition induced by phenylarsine oxide, which promotes membrane protein thiol cross-linkage in a process independent of Ca2+ or reactive oxygen species, is also strongly inhibited in acidic incubation media. In addition, we observed that the decreased protein thiol reactivity with phenylarsine oxide or phenylarsine oxide-induced swelling at pH 6.0 is reversed by diethyl pyrocarbonate, in a hydroxylamine-sensitive manner. These results provide evidence that the inhibition of mitrochondrial permeability transition observed at lower incubation medium pH is mediated by a decrease in membrane protein thiol reactivity, related to the protonation of protein histidyl residues. PMID- 10841270 TI - Selective cytostatic and neurotoxic effects of avermectins and activation of the GABAalpha receptors. AB - A natural avermectin complex, aversectin C, was shown to be capable of exerting selective cytostatic and neurotoxic effects on mammalian cells. Specifically, it killed proliferating neuroblastoma B103 cells but was non-toxic for differentiated cells of this culture. The antiproliferation action of aversectin C was not inhibited by bicuculline or picrotoxin, antagonists of the GABAalpha receptors, and was partly due to the action of avermectin A1, a component of aversectin C. Aversectin C irreversibly suppressed activity of 60% neurons in medial septal slices of the rat brain. More than 55% of them were the GABAalpha- and B1-sensitive neurons whereas the rest, about 45% neurons, were the GABAalpha insensitive and the neurotoxic effect of aversectin C was caused mainly by the B2 component. PMID- 10841272 TI - Intradermal DNA immunization: antisera specific for the membrane lectin MR60/ERGIC-53. AB - The trafficking of intracellular membrane proteins in Golgi apparatus, endoplasmic reticulum or intermediate compartment has not yet been fully elucidated. The human MR60/ ERGIC-53 and the rat p58 proteins are one such protein; and to study them in cell-free and in situ systems, high quality monospecific antisera are required. Highly specific antisera have been obtained after immunization of mice with plasmids containing a gene encoding either the full length or a truncated protein. The best results were obtained after intradermal injections of a plasmid encoding a truncated protein comprising both the luminal carbohydrate recognition domain and the stem down to a cysteine residue close to the C-terminal end, but neither the transmembrane nor the cytosolic domains. Such antisera have a very high titer and are very efficient tools to visualize the MR60 protein in situ or to selectively precipitate the MR60 proteins from a whole cell lysate. PMID- 10841271 TI - Effects of suramin on PMN interactions with different surfaces. AB - Human polymorphonuclear leukocytes (PMN) were found to tightly adhere on endothelial (lines EAhy926 and ECV304) and collagen surfaces under the influence of the chemotherapeutic drug suramin. This was observed by scanning electron microscopy and quantitated by myeloperoxidase assays. Suramin also inhibited Ca2+ ionophore A23187-stimulated leukotriene (LT) synthesis in PMN interaction with endothelial cells or with collagen surface. Suramin decreased the release of radiolabeled arachidonic acid (AA) and 5-lipoxygenase (5-LO) metabolites by prelabeled PMN stimulated with A23187. Using agents releasing the suramin stimulated adhesion namely jasplakonolide and dextran sulfate, we observed a reversal of the suramin effect on leukotriene synthesis. Jasplakonolide released the adhesion of PMN on endothelial and collagen-coated surfaces and restored 5-LO activity. Dextran-sulfate released adhesion on collagen-coated surfaces and abolished suramin inhibition. Arachidonate could also overcome adhesion and inhibition of 5-LO. We conclude that suramin-induced tight attachment of PMN on to solid surfaces lead to decreased leukotriene synthesis during subsequent A23187 stimulation in the absence of exogenous substrates. PMID- 10841273 TI - Characterization of a high-affinity folate receptor in normal and malignant human testicular tissue. AB - We have characterized the folate receptor in normal and malignant tissue from male gonads. Radioligand binding displayed characteristics typical of other folate receptors. Those included a high-affinity type of binding (K = 10(10M-1)), apparent positive cooperativity changing into non-cooperativity at low receptor concentrations, a tendency to increased binding affinity with decreasing receptor concentrations, a slow dissociation at pH 7.4 becoming rapid at pH 3.5 and inhibition by folates, in particular oxidized forms. The gel filtration profile of Triton X-100 solubilized tissue contained a 25 and 100 kDa peak of radioligand receptor. The latter peak could represent receptor equipped with a hydrophobic membrane anchor that inserts into Triton X-100 micelles. The concentration of radiolabelled receptor ranged from 0.41 nmol/g protein to 1.68 nmol/g protein in specimens of normal testicular tissue from patients with prostatic carcinomas and from 1.54 nmol/g protein to 3.82 nmol/g protein in testicular tissue from young individuals. Compared to normal testicular tissue the concentration of receptor in seminoma tissue was low (0.38-1.27 nmol/g protein) but showed a higher degree of immunoreactivity in the presence of antibodies against human milk folate binding protein as evidenced by ELISA and immunohistochemistry data. Hence a folate receptor isoform homologous to human milk folate binding protein is apparently expressed in seminomas where the total expression of receptor, however, seems to be lower than in normal testicles. PMID- 10841274 TI - Carnosine, the protective, anti-aging peptide. AB - Carnosine attenuates the development of senile features when used as a supplement to a standard diet of senescence accelerated mice (SAM). Its effect is apparent on physical and behavioral parameters and on average life span. Carnosine has a similar effect on mice of the control strain, but this is less pronounced due to the non-accelerated character of their senescence processes. PMID- 10841275 TI - Effect of ethanol on enzymatic activity of fungal laccases. AB - Blue laccase from Coriolus versicolor and blue and yellow laccases from Panus tigrinus were isolated, purified and studied in acetate buffer solutions, with and without addition of various amounts of ethanol, using syringaldazine and 2,6 dimethoxyphenol as substrates. Effect of ethanol on blue laccases could be successfully described using the mixed inhibition model, over the range of 0-2.5 M ethanol concentrations. Yellow laccase from P. tigrinus behaves differently, which may be explained by the presence of some extra molecules in its structure, which possibly stabilize the enzyme and might be exchanged in ethanol solutions. PMID- 10841276 TI - Successful transfection of lymphocytes by ternary lipoplexes. AB - Transgene expression in lymphoid cells may be useful for modulating immune responses in, and gene therapy of, cancer and AIDS. Although cationic liposome DNA complexes (lipoplexes) present advantages over viral vectors, they have low transfection efficiency, unfavorable features for intravenous administration, and lack of target cell specificity. The use of a targeting ligand (transferrin), or an endosome-disrupting peptide, in ternary complexes with liposomes and a luciferase plasmid, significantly promoted transgene expression in several T- and B-lymphocytic cell lines. The highest levels of luciferase activity were obtained at a lipid/DNA (+/-) charge ratio of 1/1, where the ternary complexes were net negatively charged. The use of such negatively charged ternary complexes may alleviate some of the drawbacks of highly positively charged plain lipoplexes for gene delivery. PMID- 10841277 TI - Cross-linking and characterisation of gelatin matrices for biomedical applications. AB - Cross-linking of gelatin A and B with N,N-(3-dimethylaminopropyl)-N'-ethyl carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) was optimised by varying the NHS/EDC molar ratio at constant EDC concentration. Native and cross linked gelatin gels were characterised using the degree of swelling, the number of free amine groups, the phase transition temperature, and titration of the carboxylic acid residues. The cross-linking reaction was most efficient at a NHS to EDC molar ratio of 0.2. At higher NHS/EDC molar ratios, the reaction of EDC with NHS becomes more pronounced, thereby reducing the effective amount of EDC for cross-linking. Swelling measurements of cross-linked gelatin gels gave deviating results when no NHS was used, which was explained by heterogeneous localisation of cross-links in the gelatin gel. The incorporation of undesired compounds into the gelatin gels during the cross-linking reaction was not observed. At optimal NHS to EDC molar ratio, gelatin A and B were cross-linked using increasing EDC/COOHgelatin molar ratios. A range of samples varying from very low cross-link density to very high cross-link density (at high EDC/COOHgelatin) was obtained. Stability of the gels is enhanced with increasing cross-link density, but a minimal cross-link density is required to obtain gelatin gels which are stable at 40 degrees C. PMID- 10841278 TI - Surface modification of the polymers present in a polysulfone hollow fiber hemodialyser by covalent binding of heparin or endothelial cell surface heparan sulfate: flow characteristics and platelet adhesion. AB - The present study addresses the problem of simultaneous surface modification of various polymers, i.e. polysulfone (PSU), polycarbonate (PC), and polyurethane (PU), which constitute the Ultraflux AV 600 S hollow fibre hemodialyser. An investigation was first made into six different chemical routes aimed at introducing carboxyl groups onto the surface of PSU, PC, and PU model polymers to which heparin (HE) or endothelial cell surface heparan sulfate (ESHS) was covalently bound via the reaction of residual amino groups and a coupling reagent. Carboxyl groups were introduced using three specific reactions based on their nucleophilic or electrophilic introduction into aromatic repeating units of the polymers and three non-specific carboxylation reactions, i.e. UV, heat or redoxactivation via nitrene or radical species. Concentrations of 1-20 nmol COOH groups per cm(-2) led to HE or ESHS surface concentrations corresponding to one or several layers. Two nonspecific carboxylation reactions followed by HE- or ESHS-coupling provided the lowest change in membrane pore structure according to cut off, clearance (urea, phosphate, maltose), ultrafiltration, and diafiltration assessments. In some cases the introduction of excess negatively-charged carboxyl groups and HE improved the flux properties of the modified membranes. The various methods were applied to the dialysis module. Platelet adhesion was not observed in the case of the ESHS-coating of PSU membrane at shear rates of 1050 s(-1), whereas HE and subendothelial matrix showed 56 and 100% coverage, respectively, under similar conditions. The coating of PSU or of other high-flux membranes by ESHS appears a promising method for improving membrane properties and to generate biocompatibility characteristics similar to those of natural blood vessels, i.e. inertness to platelet adhesion and no level effects for complement and intrinsic coagulation cascade activation. The ESHS coating may be used without anticoagulants. PMID- 10841279 TI - Synthesis and degradation of a tri-component copolymer derived from glycolide, L lactide, and epsilon-caprolactone. AB - A series of tri-component copolymers was synthesized by ring opening copolymerization of cyclic lactones, i.e. glycolide, L-lactide, and caprolactone, using stannous octoate as a catalyst. Various techniques, including FT-IR, 1H NMR, DSC, X-ray diffraction, tensile strength, and contact angle measurements, were used to elucidate structural characteristics, thermal behavior, mechanical properties, and hydrophilicity of the resulting copolymers. Data showed that the properties of these copolymers could be modulated by adjusting the composition of the copolymers. The DSC and X-ray analysis demonstrated amorphous structures for most of the PGLC copolyesters. The degradation behavior of these PGLC copolymers had been studied in vitro, i.e. in 0.10 M pH 7.4 phosphate buffer solution (PBS). The degradation was monitored by intrinsic viscosity and weight loss measurements. SEM and GPC were also used to monitor the morphology and molecular weight change during degradation. The PGLC copolymers were shown to have variable degradation rates, and most of them could disappear within a few months due to their amorphous structure and low glass transition temperature. PMID- 10841280 TI - In vitro investigation of aluminum and fluoride release from compomers, conventional and resin-modified glass-ionomer cements: a standardized approach. AB - The amount of fluoride release from dental cements necessary for an anticariogenic effect is not established: moreover, the possible toxic effects due to high fluoride and aluminum release are not well known and the results are still controversial. The aim of our study was to evaluate fluoride (F) and aluminum (Al) release from dental cements using a 'standardized approach' according to the end-use of the materials, i.e. biocompatibility testing. Two polyacid-modified resin composites of recent application, commonly called compomers (Dyract and Dyract Cem), were compared with two conventional acid-based (Fuji I, Ketac-Cem) and two resin-modified (Vitremer, Vitrebond) glass-ionomer cements (GICs). All types of cement are used in dentistry and are commercially available. Extracts of the cements into minimum essential medium, after setting over a 1-h (group A) and 1-week (group B) period, were performed. The extraction conditions were rigorously standardized. Mean values +/- standard deviation of F- and Al-levels in such extracts were measured and were expressed as microg g(-1) (micrograms of ions per gram of cement). A great difference in the amount of ion release, both F and Al, was shown among the tested materials. The GICs, as well as Ketac-Cem, released more F and Al than the compomers. All of the materials released the greatest proportion of ions when the extraction was performed in the first hour after mixing (group A). Al- and F-values showed a highly significant positive correlation, independently from the curing time. We conclude that the biological assessment of dental cements can be performed only if a pre-evaluation of the leachables is obtained by applying a standardized protocol which allows a useful comparison between the different materials. PMID- 10841281 TI - Synthesis, characterization, biodegradation, and drug delivery application of biodegradable lactic/glycolic acid polymers: I. Synthesis and characterization. AB - A series of lactic/glycolic acid polymers with various molar ratios of lactic to glycolic acid and various molecular weights were synthesized using the ring opening polymerization method. The polymerization conditions for the lactic/glycolic acid polymer synthesis were as follows: 150 degrees C, 700 microm Hg, 3 h, 0.03 wt% of catalyst (stannous 2-ethyl-hexanoate) concentration. The molecular weight of these polymers was controlled by using a molecular weight controller, lauryl alcohol. The synthesized polymers have been characterized with respect to polymer composition, molecular weight, inherent viscosity, and glass transition temperature. The characterization experiments show a good correlation between the polymer compositions and the feed ratios of lactic to glycolic acid. The molecular weight of the lactic/glycolic polymers, ranging from 10,876 to 166,630 D and the intrinsic viscosity of the polymers, ranging from 0.16 to 0.86 dl g(-1), are controlled by the amount of molecular weight controller used. The effect of the amount of the molecular weight controller on the polymer molecular weight and the polymer inherent viscosity was studied. Results indicate that the molecular weight and inherent viscosity of the polymers have a log-log linear relationship with the amount of molecular weight controller used. The lactic/glycolic acid polymers are amorphous, glassy, and transparent. The glass transition temperature of the polymers range from 21.95 to 51.29 degrees C, depending on the polymer molecular weight and the composition. PMID- 10841282 TI - Printing patterns of biospecifically-adsorbed protein. AB - The advancement of elastomeric patterning techniques in recent years has significantly enhanced our ability to spatially control biomaterial surface chemistry at the micrometre level. The application of this technology to the patterning of biomolecules onto solid surfaces has created many potential applications including the development of advanced biosensors, combinatorial library screening and the formation of tissue engineering templates. In this paper, we describe the direct patterning of protein by microcontact printing. An important consideration for the fabrication of protein micropatterns intended for these applications is the nature of the protein immobilization to a substrate. To date, the patterning of proteins by direct microcontact printing (microCP) has relied on the non-covalent adsorption to a substrate. Ideally, the proteins need to be firmly anchored onto a surface without adversely effecting their activity. Here, the high affinity avidin-biotin receptor-ligand interaction has been exploited to form arrays of avidin molecules onto a polymeric substrate expressing biotin moieties. This has created a generic technique by which any biotinylated species can be subsequently immobilized into defined patterns. Utilizing atomic force microscopy (AFM), the patterned surfaces have been characterized to molecular resolution. The micropatterned sample supported cell adhesion when biotin-(G)11-GRGDS was bound to the avidin bearing arrays. PMID- 10841283 TI - Antibody production by molecular farming in plants. AB - "Molecular farming" is the production of pharmaceutical proteins in transgenic plants and has great potential for the production of therapeutic anti-cancer antibodies and recombinant therapeutic proteins. Plants make fully functional recombinant human or animal antibodies. Cultivating transgenic plants on an agricultural scale will produce almost unlimited supplies of recombinant proteins for uses in medicine. Combinatorial library technology is a key tool for the generation and optimisation of therapeutic antibodies ahead of their expression in plants. Optimised antibody expression can be rapidly verified using transient expression assays in plants before creation of transgenic suspension cells or plant lines. Subcellular targeting signals that increase expression levels and optimise protein stability can be identified and exploited using transient expression to create high expresser plant lines. When high expresser lines have been selected, the final step is the development of efficient purification methods to retrieve functional antibody. Antibody production on an industrial scale is then possible using plant suspension cell culture in fermenters, or by the propagation of stably transformed plant lines in the field. Recombinant proteins can be produced either in whole plants or in seeds and tubers, which can be used for the long-term storage of both the protein and its production system. The review will discuss these developments and how we are moving toward the molecular farming of therapeutic antibodies becoming an economic and clinical reality. PMID- 10841284 TI - HLA-G promotes immune tolerance. AB - HLA-G is a non-classical major histocompatibility complex class I molecule that differs from the classical HLA-A, -B and -C molecules by (i) alternative splicing of mRNAs encoding for at least four membrane-bound and two soluble HLA-G isoforms, (ii) a limited polymorphism, and (iii) a tissue-restricted distribution. Studies over the past few years have elucidated the function of HLA G demonstrating inhibition of both NK cell- and T cell-mediated cytolysis. Furthermore, aside from its expression during pregnancy, we have shown that HLA-G is also expressed in solid tumor cells (i.e. human melanoma cell lines and ex vivo melanoma biopsies). Here we present a review of the current state of knowledge of the immunotolerant functions of HLA-G and their implications in materno-fetal tolerance and tumor immunosurveillance. PMID- 10841285 TI - The function of GPI-anchored proteins in T cell development, activation and regulation of homeostasis. AB - Many glycosylphosphatidyl-inositol-anchored proteins (GPI-AP) are expressed on T lymphocytes. Ligand or mAb-mediated aggregation of all GPI-AP tested to date results in the initiation of signal transduction pathways via the activation of src family protein tyrosine kinases. Src family kinases co-localise with GPI-AP in specialised sub-domains of the plasma membrane, referred to as detergent insoluble membrane microdomains (DIGS), which are thought to function as signalling platforms. GPI-AP may play a role in the regulation of T cell clonal expansion and effector functions at multiple levels, including the initiation of T cell activation through the antigen receptor complex, the regulation of ongoing responses supported by persisting antigen, as well as proliferative responses to the major T cell growth factor, IL-2. Evidence supporting the role of GPI-AP in the regulation of T cell development, activation and homeostasis is discussed, as well as insights provided by studies in humans and mice lacking GPI-AP. PMID- 10841286 TI - As2O3 induces apoptosis of the human B lymphoma cell line MBC-1. AB - OBJECTIVE: To see how arsenic trioxide (As2O3) affects proliferation of the human B lymphoma cell line MBC-1. METHODS: We studied the effect of As2O3 on MBC-1 cells and its mechanism by morphological observation, flow cytometry assay and DNA gel electrophoresis. RESULTS: As2O3 could upregulate p53 gene expression at protein level, inducing cell apoptosis and inhibiting the proliferation of MBC-1 cells. Upregulation of p53 expression appears to be important in the apoptosis of MBC-1 cells. CONCLUSIONS: As2O3 can inhibit the proliferation of MBC-1 cells by upregulating p53 gene expression, thus inducing apoptosis. PMID- 10841288 TI - CD95L. PMID- 10841287 TI - Correlation between p53 gene mutations and p53 protein accumulation evaluated by different methodologies. AB - Mutations in the p53 gene are the most common genetic alterations in many tumour histotypes. Many of these mutations induce conformational changes resulting in p53 protein stabilisation and consequently an accumulation detectable with immunochemical methods. Available data on the correlation between p53 gene alterations and p53 overexpression widely vary. In this study we analysed the correlation between p53 gene alterations detected by DGGE, SSCP and sequencing and protein expression detected by flow cytometric and immunohistochemical approaches by using PAb 1801 antibody. The study was performed on 21 bladder tumours and 10 cell lines derived from different tumour histotypes as representative of different methodologic problems which can be met starting from different types of biological material. The best correlation (81%) was observed between p53 mutations and FCM results, using a double evaluation criterion for the latter which includes the percentage of positive cells and "delta values", evaluated as the difference between the mean values of Pab 1801 stained cells and isotypic control. The high correlation obtained between results from this FCM double criterion and p53 gene mutations is a good starting point for the analysis on large series of tumours and for a multiparameter FCM analysis including p53 protein levels. PMID- 10841289 TI - CD121a. PMID- 10841290 TI - CDw121b. PMID- 10841291 TI - CDw137. PMID- 10841292 TI - CD153. PMID- 10841293 TI - CD154. PMID- 10841294 TI - Preventing and reducing substance use among institutionalized adolescents. AB - The Residential Student Assistance Program, serving high-risk, multiproblem, inner-city, primarily African-American and Latino youth, was evaluated for its ability to prevent and decrease alcohol and other drug use. Participants were drawn from several adolescent residential facilities: three foster care sites for abused, neglected, orphaned, or troubled adolescents, a nonsecure facility for adjudicated juvenile offenders, a treatment center for teens with severe psychiatric problems, and a locked county correctional facility. In addition, comparison groups were employed. A 5th-year outcome evaluation documented the program's effectiveness in both preventing and reducing substance use among participants, with impact related to program dosage. Qualitative process data clarified and strengthened confidence in the quantitative outcomes. PMID- 10841295 TI - Parental involvement with adolescents' education: do daughters or sons get more help? AB - This research examined whether parents were involved differently with the education of their adolescent daughters and sons. The investigation used data from the National Education Longitudinal Study (NELS), which collected information from approximately 25,000 eighth-grade students. Several types of parental involvement were analyzed for gender differences, including school discussion, parent-school connection, parental expectations, parental attendance at school events, and three measures of parental supervision (checking homework, limiting television watching, and limiting going out with friends). The results showed that, net of students' grades, tests scores, and educational aspirations, parents helped daughters in some ways and sons in other ways. Generally, daughters experienced more parental involvement with their education than did sons. The findings are discussed in terms of parents' traditional socialization practices versus a shift in parental treatment in response to social trends. PMID- 10841296 TI - The transition to high school for academically promising, urban, low-income African American youth. AB - In nine urban Ohio school systems, low-income minority students identified as academically promising in sixth grade are eligible to participate in an intervention program. In the present study, twenty-two African American students in the program were asked to provide their perceptions of the transition to ninth grade. Specifically, the role of motivating factors, peers, school, teachers, parents, and neighborhood were examined. These students faced similar stressors, yet some were more able to achieve academic success. Results highlight the salience of mothers, the challenges of the ninth-grade curriculum, and adjustment to a bigger, more complex school environment for high and low performers. The implications for improving cooperation between school and family are discussed. PMID- 10841297 TI - The moral justification scale: reliability and validity of a new measure of care and justice orientations. AB - Research increasingly suggests that there are limitations to Kohlberg's theory of moral development. Gilligan in particular has observed that Kohlberg's theory considers abstract principled reasoning as the highest level of moral judgment, and penalizes those who focus on the interpersonal ramifications of a moral decision. Gilligan calls these justice and care orientations. The present paper describes the development of the Moral Justification Scale, an objective measure of the two orientations. The scale consists of six vignettes, of which two are justice oriented, two are care oriented, and two are mixed, incorporating both orientations. Construct validity was evaluated by expert judges and, overall, was high. Cronbach's alpha was .75 for the Care subscale and .64 for the Justice subscale, indicating adequate internal consistency. Split-half reliabilities were as follows: Care, r = .72, p < .01, and Justice, r = .60, p < .05. Regarding test retest reliability (approximately two weeks), r = .61, p < .05, for Care; r = .69, p < .05, for Justice. Neither subscale correlated significantly with the Marlowe-Crowne Social Desirability Scale. Thus, the Moral Justification Scale shows promise as an easily administered, objectively scored measure of Gilligan's constructs of care and justice. PMID- 10841298 TI - The school adjustment of South Asian immigrant children in the United States. AB - This study examined the school adjustment process among South Asian children who had immigrated to the United States with their parents, and who had below-average grades. Both risk and protective factors for dropping out of school were explored in the context of the traditions, familial values, and social norms of South Asians. Data were collected from 75 parents and 75 children in separate semistructured interviews. Content analysis revealed three major themes: congruence of the parents' and school's views on the value of education, congruence of the parents' and children's beliefs that education is the tool to achieve goals, and determination of the children to achieve goals. The low level of proficiency in English was found to be a critical factor in low achievement and school failure. It was concluded that parental encouragement to succeed, in conjunction with teachers' efforts, can be used to facilitate children's school adjustment. Strategies for assisting immigrant children are discussed. PMID- 10841299 TI - Prenatal maternal predictors of cognitive and emotional delays in children of adolescent mothers. AB - The present study examined relationships among prenatal characteristics of 121 adolescent mothers-including cognitive readiness for parenting, intelligence, social support, and personal adjustment-and intellectual-linguistic development, social-emotional functioning, and adaptive behavior in their children at three years of age. Only 28% of the children scored within normal ranges on all three types of outcomes. Intellectual and linguistic delays were predicted best by prenatal measures of maternal Performance IQ and social support from extended family. Socioemotional problems were predicted best by maternal internalizing problems and social support from partner and friends. Adaptive behavior was associated with parenting style. Implications for the early identification of high-risk children--and associated intervention programs--are discussed. PMID- 10841300 TI - Sampling issues in research on adult children of alcoholics: adolescence and beyond. AB - A critical examination of 98 studies on adult children of alcoholics, published from 1986 to 1995, was prompted by suggestions that sampling issues confound research findings. Over half of the studies that were reviewed involved college students. It is important to understand the methodology used in creating a knowledge base that relies heavily on late adolescent and young adult offspring of alcoholics. Thus, two main questions were asked when reviewing these studies: What methods were employed to classify offspring as being children of alcoholics? To what extent does our knowledge about adult children of alcoholics depend on college student samples and clinical samples? PMID- 10841301 TI - Assessing African American adolescents' risk for suicide attempts: attachment theory. AB - The threat of separation from a parent theoretically increases the risk of adolescent suicide attempts. The present study evaluated this and other hypothesized risk factors in a sample of adolescent suicide attempters and nonsuicidal controls, using the Psychiatric Consultation Checklist (Lyon, 1987). Stepwise logistic regression was used to predict group membership. It was found that threat of separation from a parental figure, insomnia, neglect, substance abuse, suicidal ideation, and failing grades were the strongest predictors of suicide attempt. Ten predictor variables correctly identified 97% of suicide attempters and 86% of nonattempters. Unexpected findings included high levels of truancy, threatening others, and separation from a parent before the age of 12 among nonattempters. PMID- 10841302 TI - Differences between fathers and mothers in the treatment of, and relationship with, their teenage children: perceptions of Chinese adolescents. AB - Chinese adolescents' perceptions of differences between mothers and fathers in parenting styles, parent-adolescent communication (frequency and related feelings), and quality of the parent-adolescent relationship were assessed via questionnaires and individual interviews. Fathers, as compared with mothers, were perceived to be less responsive, less demanding, to demonstrate less concern, but to be more harsh, and paternal parenting was less liked. There was less communication with fathers, and adolescents reported more negative feelings when communicating with fathers than with mothers. They evaluated the father adolescent relationship more negatively than they did the mother-adolescent relationship. Adolescent females, as compared with males, perceived their parents to be more demanding but less harsh. Parenting characteristics were rated less favorably across time. PMID- 10841303 TI - Identity status formulae: generating continuous measures of the identity statuses from measures of exploration and commitment. AB - Mathematical formulae were devised for the purpose of generatin continuous measures of the four identity statuses from measures of exploration and com measures of exploration and commitment. The formulae were consistent with the conceptual definitions of the statuses. They were found to be effective both in terms of generating continuOUs measures of the statuses and in terms of deriving status assignments. PMID- 10841304 TI - The myth of peer pressure. AB - The construct of peer pressure was examined as part of a qualitative study of the determinants of mental health for 41 high-risk adolescents. While the concept of peer pressure enables adults to explain youths' troubling behaviors, content analysis of the participants' accounts of their lives revealed peer pressure to be a myth. The youths indicated that adoption of the behavior and appearance of peers was a consciously employed strategy to enhance personal and social power. Association with peers was used to construct and maintain health-promoting identities that challenged the stigmatizing labels given to them by others. Three developmental stages to this process of identity construction were identified. During stage one, vulnerable youths learn to maintain a singular self-definition through interaction with peers. In stage two, youths purposefully use their peer relations to experiment with multiple identities. During stage three, youths collaborate with peers as equal partners in the construction of one or more identities for which they find acceptance. PMID- 10841305 TI - The child and adolescent functional assessment scale (CAFAS): a dynamic predictor of juvenile recidivism. AB - This study tested the degree to which the Child and Adolescent Functional Assessment Scale (CAFAS; Hodges & Wong, 1996), a mental health assessment tool, predicted recidivism among juvenile offenders. The CAFAS, which is sensitive to rehabilitation treatment effects, was compared with factors insensitive to rehabilitation (e.g., age, ethnicity, sex, and number of prior offenses). Also addressed was the methodological issue of whether to treat recidivism as a continuous or a dichotomous variable. The CAFAS was found to be significantly related to recidivism. The practical application of the results, as well as the value of emphasizing research on dynamic predictors that enable policy makers to target at-risk juveniles, is discussed. PMID- 10841306 TI - Repeat pregnancy among urban adolescents: sociodemographic, family, and health factors. AB - First-time adolescent mothers are at high risk for a repeat pregnancy. The present investigation, part of an ongoing longitudinal study, examined sociodemographic, family, and health factors associated with repeat Pregnancy in a clinic sample of urban, first-time adolescent mothers (ages 13 to 17 years). They were predominantly African-American and from low-income households. Repeat pregnancy within one year and between one and two years postpartum was determined from medical records. Summary statistics, point biserial correlations, and chi square statistics were used to analyze the data. Results indicated that postpartum contraceptive method was associated with repeat pregnancy at Year 1; contraceptive use, maternal age, history of miscarriages, and postpartum contraceptive method were associated with repeat pregnancy at Year 2. It was concluded that efforts to prevent repeat pregnancies among first-time adolescent mothers should include the continuous monitoring of contraceptive use, as well as the promotion of long-acting contraceptives (e.g., medroxyprogesterone or progesterone implants). Further, counseling should be offered to adolescent mothers with a history offered miscarriages. PMID- 10841307 TI - Relationships between anxiety, fear, self-esteem, and coping strategies in adolescence. AB - This study investigated the relationships between anxiety, fear, self-esteem, and coping strategies in a sample of 224 postprimary students (years 7, 9, and 12) in Australia. In particular, it sought to determine whether there were any significant changes between years 7 and 12 and, if so, whether these changes were gender specific. The results indicated that the girls had consistently low levels of self-esteem. The boys showed a significant decrease in both anxiety and fear by year 12. For the coping strategies, a three-factor solution accounted for 64.2% of the variance. Finally, the findings suggested that, by year 12, boys and girls were using different coping strategies, with boys more successfully reducing both fear and anxiety. PMID- 10841308 TI - Histoenzymology of oxidases and dehydrogenases in peripheral blood lymphocytes and monocytes for the study of mitochondrial oxidative phosphorylation. AB - Histoenzymological methods usually performed on muscle fibres have been adapted to assess the functioning of oxidative phosphorylation in human circulating blood lymphocytes and monocytes. Oxidases and dehydrogenases were analysed in lymphocyte/monocyte smears. The specificity of each histoenzymological reaction was tested using a specific respiratory chain inhibitor: rotenone for NADH diaphorase, thenoyltrifluoroacetone for succinate dehydrogenase, potassium cyanide for cytochrome c oxidase and oligomycin for ATPase. Complex I activity was detected, but inhibition with rotenone was incomplete. Complexes II, IV and V were almost completely inhibited. These observations indicate that histoenzymology is a valuable method for detecting the activity of these oxidative phosphorylation enzymes in lymphocytes and monocytes. The histoenzymology tests performed on fresh peripheral blood cells resembled those used for muscle biopsies. They could be useful for the diagnosis of respiratory chain disorders in patients. PMID- 10841309 TI - Levels, phosphorylation status and cellular localization of translational factor eIF2 in gastrointestinal carcinomas. AB - The level of expression and the phosphorylation status of the alpha subunit of initiation factor 2 (eIF2alpha) protein have been determined by comparing samples from human stomach, colon and sigma-rectum carcinomas with normal tissue from the same patients. The unphosphorylated and phosphorylated levels of cytoplasmic eIF2alpha, as well as the percentage of phosphorylated factor over the total, were significantly higher in stomach, colon and sigma-rectum tumours compared with normal tissue. The expression of this factor was also studied by using immunocytochemical methods, where redistribution towards the nucleus in tumour cells as compared with normal tissue was observed. Our results support a likely implication of eIF2alpha in gastrointestinal cancer. PMID- 10841310 TI - Ito cell morphology, alpha-smooth muscle actin and collagen type IV expression in the liver of patients with gastric and colorectal tumors. AB - The alteration in sinusoidal collagen type IV occurrence, and myofibroblastic (alpha-SMA-positive) Ito cellular transformation are described in the liver of patients with malignant gastric and colorectal tumors, using electron microscopy as well as light microscopical and ultrastructural immunohistochemistry. The ultrastructural finding revealed transformation of Ito cells mostly into transitional cells in highly differentiated primary tumors and into transitional and myofibroblast-like cells with expressed changes in the other sinusoidal cells in poorly differentiated tumors. Ito cell numbers increased significantly in the livers of cancer patients. A highly significant statistical association was obtained between Ito cell numbers on the one hand and collagen type IV and alpha SMA immunoreactivity on the other hand in the pericentral zone of the liver lobule. Ultrastructural immunohistochemistry showed increased collagen IV immune deposits in the space of Disse, assembled for the most part around and inside transitional cells. Alpha-SMA immunoreactivity was detected in activated Ito cells diffuse in the lobule, with stronger expression in the intermediate and pericentral zones. It is suggested that stimuli which can influence Ito cell transformation are produced by tumor cells from the primary tumor (TGF-beta1, TNF alpha, PDGF-beta etc.) and from the metastasizing gastric or colorectal tumor cells--matrix metalloproteinase-2 (MMP-2). It is suggested that sinusoidal extracellular matrix deterioration creates a barrier for cancer invasion on the one hand, or possibly facilitates metastasizing by ensurance of matrix for adhesion on the other hand. PMID- 10841311 TI - Confocal analysis of cytoskeletal organisation within isolated chondrocyte sub populations cultured in agarose. AB - This study reports the cytoskeletal organisation within chondrocytes, isolated from the superficial and deep zones of articular cartilage and seeded into agarose constructs. At day 0, marked organisation of actin microfilaments was not observed in cells from both zones. Partial or clearly organised microtubules and vimentin intermediate filaments cytoskeletal components were present, however, in a proportion of cells. Staining for microtubules and vimentin intermediate filaments was less marked after 1 day in culture however than on initial seeding. For all three cytoskeletal components there was a dramatic increase in organisation between days 3 and 14 and, in general, organisation was greater within deep zone cells. Clear organisation for actin microfilaments was characterised by a cortical network and punctate staining around the periphery of the cell, while microtubules and vimentin intermediate filaments formed an extensive fibrous network. Cytoskeletal organisation within chondrocytes in agarose appears, therefore, to be broadly similar to that described in situ. Variations in the organisation of actin microfilaments between chondrocytes cultured in agarose and in monolayer are consistent with a role in phenotypic modulation. Vimentin intermediate filaments and microtubules form a link between the plasma membrane and the nucleus and may play a role in the mechanotransduction process. PMID- 10841312 TI - Gene expression and immunohistochemical localization of biglycan in association with mineralization in the matrix of epiphyseal cartilage. AB - This study has used in situ hybridization, Northern blot analysis, and immunohistochemistry at the light and electron microscope levels to localize mRNAs and core proteins of biglycan in developing tibial epiphyseal cartilage of 10-day old Wistar rats. The expression of mRNAs and core proteins of biglycan appeared prominent in hypertrophic and degenerative chondrocytes associated with the epiphyseal ossification centre and the growth plate cartilage, but was not seen in the rest of epiphyseal cartilage. Northern blot analysis confirmed biglycan mRNA expression in the epiphyseal cartilage. Ultrastructural immunogold cytochemistry of the growth plate revealed that prominent immunolabelling was confined to the Golgi apparatus and cisternae of rough-surfaced endoplasmic reticulum of the hypertrophic and the degenerating chondrocytes, the early mineralized cartilage matrices of the longitudinal septum of the lower hypertrophic and the calcifying zones, and fully mineralized cartilage mitrices, which were present in the metaphyseal bone trabeculae. Furthermore, Western blot analysis of biglycan in extracts of fresh epiphyseal cartilage revealed that an EDTA extract, after chondroitinase ABC digestion, contains core proteins of biglycan, indicating the presence of biglycan in mineralized cartilage matrices. These results indicate that the distribution of biglycan is associated with cartilage matrix mineralization. PMID- 10841313 TI - Expression of N-acetylglucosamine residues in developing rat fundic gland cells. AB - The development of rat fundic gland was studied by immunohistochemistry using a recently developed monoclonal antibody, HIK 1083, at both light and electron microscope levels. Antibody HIK 1083 recognized oligosaccharides with a non reducing terminal alpha-linked N-acetylglucosamine (GlcNAc) residue. In the developing rat fundic gland, cells expressing alpha-GlcNAc residues were discernible from day 19.5 of gestation and continued to exist till adult. The distribution of the alpha-GlcNAc expressing cells was consistent with that described previously for cells reacting to Griffonia simplicifolia lectin (GSA II) in all developmental stages. These cells were located at the bottom of the fundic gland when they first appeared. With the elongation and maturation of the gland, these cells moved upwards and were finally restricted in the neck region of the gland. Combining previous reports and the present electron microscopical observations, HIK 1083-positive cells in the adult rat fundic gland are mucous neck cells. The interaction between antibody HIK 1083 and GSA-II lectin was investigated. GSA-II prevented the subsequent binding of HIK 1083, while HIK 1083 did not prevent GSA-II binding to mucous neck cells. Our results suggested that alpha-GlcNAc residues exist in rat fundic gland from day 19.5 of gestation and continue to exist till adult. Cells expressing alpha-GlcNAc residues appeared as typical mucous neck cells from postnatal four weeks. PMID- 10841314 TI - A guest editorial: stump the professor. PMID- 10841315 TI - BRCA1 and BRCA2 gene mutations: decision-making dilemmas concerning testing and management. AB - The identification of the BRCA genes, and their possible etiologic relationship with various forms of inherited cancer, has been recognized universally as a cornerstone in the search of cancer's genetic susceptibility. Female BRCA gene mutation carriers are found to carry an increased risk of developing breast or ovarian cancer and to a lesser degree, colon cancer, and male BRCA mutation carriers are also related to an increased risk of breast, colon, or prostate cancer. Although genetic testing promises a possible future presymptomatic determination and treatment of women who are genetically susceptible to cancer, current data reveal certain dilemmas and uncertainties regarding our ability to interpret the results from testing and offer effective management options. In addition, several complex ethical, legal, and social issues have been revealed with the advent of this new information, which also confirm the need for additional research regarding the most effective use of this genetic information and for the establishment of appropriate clinical management strategies. PMID- 10841316 TI - Conservative medical management of advanced cervical ectopic pregnancies. AB - Cervical ectopic pregnancy is the rarest form of ectopic gestation. Viable cervical ectopic pregnancies of 10 weeks' gestation or more are even rarer. It is unclear whether these advanced cervical ectopic gestations should be managed primarily by surgical evacuation or by more conservative medical management with chemotherapeutic agents. Cases of medical treatment of viable cervical ectopic gestations reported in the literature are reviewed. An additional case of a 10.6 weeks of gestation, viable cervical ectopic pregnancy treated with methotrexate, intrafetal potassium chloride, and intramuscular methotrexate is added to the previously reported literature. Advanced cervical ectopic pregnancies with fetal cardiac activity at 10 or more weeks' gestation may be successfully managed with chemotherapeutic agents. Ultrasound-guided intrafetal injection of feticidal agents may be preferable to maternal systemic chemotherapy alone. PMID- 10841317 TI - Endometrial-myometrial interface: relationship to adenomyosis and changes in pregnancy. AB - The endometrial-myometrial interface (EMI) is an important region of the human uterus, which has attracted little research attention. This mucosal-muscular interface has characteristic features when compared with other similar interfaces in the human body. It lacks an intervening tissue layer and as a result, the endometrium sits directly on the myometrium rendering it vulnerable to invasion by the endometrium. Both endometrium and myometrium are sensitive to sex steroids, and their structure and function depend to a large extent on the sex hormonal milieu. Endometrium, which forms one border of the EMI, is a complex tissue consisting of several polarized microenvironments. At a cellular level, sex steroids interact with local mediators secreted by a variety of cell types and are important in maintaining the complex structure and function of the endometrium. Basal endometrium contains prominent aggregates of leukocytes that may be important in controlling local cell growth and function. Myometrium also has a distinct zonal anatomy. The recently described junctional zone differs structurally and functionally from the outer myometrium, although these functions are not yet clearly understood. Embryologically, it originates from mullerian ducts together with endometrium, whereas the outer myometrium has a non-mullerian origin. During early pregnancy, the EMI is disturbed by invading trophoblast. Alterations of myometrial intercellular matrix proteins together with expression of appropriate receptors by the trophoblast seem to regulate this unique interaction. The EMI also is disrupted in adenomyosis. The sequence of events taking place at the EMI during development of this pathology is still debated. PMID- 10841318 TI - Thiorphan, an inhibitor of neutral endopeptidase/enkephalinase (CD10/CALLA) enhances cell proliferation in bone marrow cultures of patients with acute leukemia in remission. AB - Thiorphan, (DL-mercapto-2-benzylpropanoyl)-glycine is a potent and specific inhibitor of membrane metallo-endopeptidase (EC 3.4.24.11, CD10). We explored its effects in short-term clonal cultures of the bone marrow from 10 patients with acute leukemia in remission. The cell suspensions were incubated with thiorphan (10(-13) to 10(-5) M) and seeded for the granulocyte/macrophage-colony forming unit (GM-CFU) assay. In normal bone marrow samples the median seeding efficiency was 119 colonies and clusters per 10(5) cells and thiorphan caused slight stimulation of the clonal growth in concentrations above 10(-9) M. In the leukemic samples, the median seeding efficiency varied from 10 to 366 colonies and clusters per 10(5) seeded cells. Meaningful alterations of the clonal growth were noted in 32 out of 83 thiorphan-treated cultures (39%). In those 32 cultures the stimulatory effects outnumbered the inhibitory effects (24 versus 8). Thus, thiorphan stimulated the progenitor cell proliferation in bone marrow samples from the normal donor and from the patients with acute leukemia in remission. Thiorphan binding to CD10 might interfere with the processing of neuropeptide hemoregulatory factors and thus influence the progenitor cell proliferation. PMID- 10841319 TI - A practical quick staining method using hydrochloric acid-fast metachromatic dye for megakaryocytes. AB - A specific stain using violet polymethine dye (VPM stain) for megakaryocytes was first developed by Kass (1995). We have modified this method for practical use in bone marrow specimens. The modified VPM stain labels megakaryocytes very well, while other marrow cells are poorly colorized. This staining procedure was more stable, and its color intensity was finer and clearer than the original. Using this stain, morphologic classification of megakaryocytes in bone marrow specimens from 11 normal and 8 myelodysplastic syndrome (MDS) patients was performed. Many megakaryocytes observed in MDS patients were juvenile compared with normal subjects according to their morphology. Blasts from acute megakaryoblastic leukemia (M7) and from a megakaryoblastic cell line (Mo7e) were also clearly stained with our method. This staining method is practical and very useful for rapid identification of megakaryocyte distribution and morphology. PMID- 10841321 TI - Serum L-selectin and P-selectin levels in lymphomas. AB - The migration of normal and malignant lymphoid cells is governed by specific adhesion molecules. Selectins comprise a family of adhesion receptors expressed by leukocytes, platelets and endothelial cells. In this study, the serum levels of soluble L-selectin and P-selectin were measured in patients with non-Hodgkin's lymphoma and Hodgkin's disease and found to be significantly elevated in both patient groups compared to healthy controls. This result provides evidence that alterations in the expression and function of adhesion molecules may play an important role in the progression of lymphomas. Further studies are awaited to establish the exact roles of these adhesion molecules in distinct patterns of growth and spread of lymphomas. PMID- 10841320 TI - Lymphoma in Castleman's disease, acute lymphocytic leukemia, adult T-cell leukemia and cutaneous T-cell lymphoma accompanied with high serum soluble Fas ligand levels. PMID- 10841322 TI - Serum IL-18 levels in patients with various hematological disorders. PMID- 10841323 TI - Hypercoagulability in various autoimmune diseases: no association with factor V Leiden mutation. AB - The coagulation factor V Leiden mutation, leading to resistance to activated protein C (APC), is the most common inherited risk factor for venous thrombosis. In various systemic autoimmune diseases the hypercoagulable state was shown to be associated with the presence of antiphospholipid antibodies (aPL). Our aim was to determine the prevalence of both, Leiden mutation and aPL in autoimmune diseases and their impact on the occurrence of venous thrombosis. The dataset consists of results from 137 patients having Sjogren's syndrome (n = 50), progressive systemic sclerosis (n = 43) (PSS), undifferentiated connective tissue disease (n = 24) (UCTD) and mixed connective tissue disease (n = 20) (MCTD) with or without venous thromboembolic complications. The Leiden mutation was detected with polymerase chain reaction (PCR), aPL, such as lupus anticoagulant (LA) with screening and confirmatory procedures and others with enzyme linked immunosorbent assay (ELISA). The prevalence of mutation ranged between 8.3% and 18.0% (13.1%). The thromboembolic risk was found to be increased in the presence of aPL. Eight patients (5.84%) (4 heterozygous) experienced thromboembolic events and 3 out of 4 heterozygous showed aPL positivity, too. There were no difference between the frequencies of Leiden mutation in examined systemic autoimmune diseases and unselected populations. PMID- 10841324 TI - Simultaneous appearance of mycosis fungoides and chronic lymphocytic leukemia in the same patient. AB - A patient is presented having simultaneously chronic lymphocytic leukemia with a monoclonal B-lymphocyte population and mycosis fungoides with atypical T-cell population in the skin lesion and in the enlarged lymphoid nodes confirmed by detailed phenotyping. PMID- 10841325 TI - Congenital dyserythropoietic anemia type I with ringed sideroblasts. AB - A 25-year-old male with anemia, jaundice and liver dysfunction was admitted to our institution. Anisopoikilocytosis with tear-drop forms, polychromasia, basophilic stippling in peripheral blood smear, erythroid hyperplasia with megaloblastoid changes, binucleated cells and intranuclear bridging in bone marrow aspirate and spongy, unevenly condensed nuclear chromatin in electron microscopy studies indicated that he had congenital dyserythropoietic anemia (CDA) type I. As a rare finding in CDA, ringed sideroblasts were noted. It is proposed that this patient is an example for the designation 'variant congenital dyserythropoietic anemia with ringed sideroblasts'. PMID- 10841326 TI - Recurrent venous thrombosis in a patient with chronic lymphocytic leukemia and acquired protein S deficiency. AB - A patient with chronic lymphocytic leukemia and an undetectable plasma level of protein S (PS), associated with recurrent venous thrombosis, is described. The laboratory investigation revealed the concomitant presence of an inhibitor directed to PS and a monoclonal protein in the patient's plasma. After treatment with prednisone and cyclophosphamide both the inhibitor to PS and the monoclonal component disappeared. PMID- 10841327 TI - Severe amyloidosis with mild multiple myeloma--an unusual course. AB - Amyloidosis may be primary or myeloma-associated. Skeletal lesions and the percentage of bone marrow plasma cells (<10% in primary, >20% in myeloma) account for the major differences between the two varieties. In the literature there are rare cases of primary amyloidosis presenting without myeloma and followed by development of myelomatous manifestations. Usually, the primary disease (i.e. the myeloma) is advanced, when amyloidosis is diagnosed. We describe a patient who had presented with a severe and progressive systemic amyloidosis and was diagnosed later to have a mild light chain myeloma. Aggressive treatment with melphalan, prednisone and colchicine resulted in a temporary partial remission, followed by a rapid downhill course, and the patient's death. The point of relatively mild myeloma following a rapidly progressive course of advanced amyloidosis is emphasized. Awareness of the possibility of such a combination may lead to early diagnosis, a more aggressive or novel therapeutic approach and, possibly, to a better prognosis. PMID- 10841328 TI - Co-existence of cutaneous true histiocytic lymphoma with refractory anaemia: report of a case. AB - The case of a 70-year-old man with refractory anaemia who developed cutaneous true histiocytic lymphoma two years after diagnosis, is described. Whether this association is a simple coincidence or indicates some etiopathogenetic relationship between these two unrelated disorders is unknown and needs the analysis of more cases. PMID- 10841330 TI - Weak beam under convergent beam illumination AB - The weak beam technique is now used widely for the determination of stacking fault energies, in particular for intermetallic alloys, and the accuracy of the approach is critically dependent upon the reliability of the relationship between the image and the actual position of the dissociated dislocations. Examining as a model case a dislocation dissociated into two Shockley partial dislocations in Cu at 100 kV for orientations ranging through the g(3g) weak beam condition, image simulations are used to explore the accuracy to which the true spacing between the partial dislocations can be determined from the spacing measured on the image as a function of the dislocation character, the foil thickness, the dislocation depth in the foil, the diffraction condition and the beam convergence. It appears that for image simulations and for the given conditions a beam convergence of about 5 mrad allows to greatly improve the accuracy, and that beam convergence must be taken into account quantitatively when deducing the true partial dislocation spacing as it is the principal parameter controlling the precision in this type of measurement. PMID- 10841329 TI - Primary pseudomonas meningitis in an adult, splenectomized, multitransfused thalassaemia major patient. AB - A 19-year-old splenectomized, multitransfused female patient with beta thalassaemia major developed primary meningitis due to P. putida. Her blood cultures were negative. P. putida is an unusual nosocomial organism to cause primary meningitis. Infection due to this organism carries high mortality. However, owing to early diagnosis and energetic treatment this patient survived without any sequelae. A review of serious infections over the last 7 years in patients in our thalassaemia care centre revealed 11 serious infections among our splenectomized patients (n = 46) and none in the non-splenectomized group (n = 106). Surprisingly, all overwhelming infections (23.8% in the splenectomized group) were caused by Gram-negative bacilli like Klebsiella, Pseudomonas, Aeromonas and Campylobacter species. As all our splenectomized patients had prior pneumococcal vaccination and oral penicillin prophylaxis, overwhelming septicaemia due to S. Pneumoniae was successfully prevented, but an increasing incidence of overwhelming sepsis due to Gram-negative bacilli, against which no vaccination or suitable prophylactic antibiotics are available, is now posing a new threat to this vulnerable group of patients. PMID- 10841331 TI - Quantification of high-resolution electron microscope images of amorphous carbon AB - Quantitative comparisons of experimentally obtained and simulated high-resolution electron microscope images have shown that the contrast in experimental images is usually much less than is predicted by simulations. The aim here is to investigate this loss of contrast as a function of image spatial frequency using high-resolution images of amorphous carbon. It seems that experimental images of amorphous carbon have an unexpectedly high contrast for the low spatial frequencies but that the loss of contrast is constant for frequencies above 0.5 nm(-1). PMID- 10841332 TI - Atom pair persistence in disordered materials from fluctuation microscopy AB - We show that the kinematical theory of fluctuation microscopy for a disordered specimen can be reformulated in terms of an atom pair-pair correlation function. Resolution and coherence are two experimental variables which yield a two dimensional fluctuation map. This map is a transformation of the pair-pair correlation function, but can be directly interpreted in terms of a pair persistence function (PPF). We show that the PPF reveals the correlation length and structure for a simple paracrystalline model. PMID- 10841333 TI - Structure analysis by diffraction of amorphous zones created by Ni ion implantation into pure Al AB - The implantation of Ni ions into pure Al leads to the formation of approximately 10 nm amorphous zones (AZ) which induce diffuse rings in the diffraction pattern in addition to the diffraction spots of the crystal. Measurements by energy dispersive spectrometry and electron energy loss spectrometry attributed to the amorphous zones an average Ni concentration of 25 at%. The exact structure of these AZ is still unknown. The structure is characterized here by both the total and partial radial distribution functions (RDF). Structure factor deduced from experiments is compared to calculated one. For this purpose, molecular dynamic (MD) simulations are used to model the AZ structure. The RDF are determined using this structure and analytical calculation of the diffraction pattern is achieved. Simulations of the diffraction pattern of the simulated MD sample using both a kinematic and a dynamic approach are achieved to refine the analytical procedure used on the experimental diffraction patterns. It appears that the amorphous structure is well reproduced by the MD simulations. Analytical calculation reveals the presence of a well-established chemical order in the amorphous material. PMID- 10841334 TI - The development of Fresnel contrast analysis, and the interpretation of mean inner potential profiles at interfaces AB - This paper provides a summary of recent published and unpublished research on the development of Fresnel contrast analysis, a transmission electron microscopy technique for measuring the mean inner potential profile across an interface or a narrow layer. An algorithm for finding a best-fitting potential profile is described, energy-filtered experimental data are analyzed and contributions to Fresnel contrast from surface grooves and space charge are assessed. Many of the conclusions drawn are equally relevant for the interpretation of phases measured using off-axis electron holography. PMID- 10841335 TI - Measurement of roughness and diffuseness of interfaces AB - We propose a simple formalism which allows the separation of the contributions, due to roughness and chemical interdiffusion, to the total width of an interface by using two-dimensional information from either images or chemical maps. A definition is also proposed for the roughness of an interface in terms of iso concentration surfaces. The formalism is based on the relation between projection in real space and the corresponding section in Fourier space, the main hypotheses being that the interface roughness is isotropic and that the composition profile is constant along the interface. The method, although general, will be illustrated with results on Fresnel imaging of Cu-Co magnetic multilayers. PMID- 10841336 TI - Trace analyses from LACBED patterns AB - Trace analysis is of major interest in transmission electron microscopy since the identification of the (h k l) indices of lattice planes or the [uvw] indices of lattice directions is required for the complete analysis of crystal defects (stacking faults, dislocations, etc). It is usually carried out from observations of micrographs and corresponding selected area electron diffraction patterns. The main difficulty comes from the rotation occurring between the image and the diffraction pattern. Therefore, the method requires a careful calibration of this rotation. The LACBED patterns have a unique property: they display information connected both with the direct and the reprocal lattices. The shadow image of the illuminated area of the specimen (direct lattice) is superimposed with the LACBED pattern composed of Bragg lines (reciprocal lattice). Since this shadow image is not rotated (or rotated by 180 degrees C) with respect to the diffraction pattern, LACBED patterns can be conveniently used to identify planes and directions. Several experimental methods are described. Most of them require observation of Bragg lines which are parallel or perpendicular to the trace of the analysed plane or direction. PMID- 10841337 TI - The dependence of equilibrium film thickness on grain orientation at interphase boundaries in ceramic-ceramic composites AB - High-resolution transmission electron microscope observations of hexagonal boron nitride - 3C silicon carbide interphase boundaries suggest that where one or more phases is highly anisotropic, an orientation dependence on equilibrium film thickness can arise. Theoretical considerations of this phenomenon in terms of the equilibrium thickness of an amorphous film between two crystalline media are consistent with the trend seen experimentally. PMID- 10841338 TI - Use of the track structure approach in TEM AB - Track structures can be usefully used to understand concepts and measurements not adequately treated by conventional scattering theory using the Born approximation. An example is the impact parameter used experimentally in energy loss and energy-dispersive spectroscopies in the transmission electron microscope. An anomaly is discussed which derives from the wave-particle duality in quantum mechanics in the context of near relativistic speeds encountered in the electron microscope. PMID- 10841339 TI - List of publications of William Michael Stobbs PMID- 10841340 TI - A useful in vitro blood-brain barrier model. PMID- 10841341 TI - Endogenous APP derivatives oppositely modulate apoptosis through an autocrine loop. AB - We have recently shown that, in rat cerebellar granule cells, apoptosis triggered by KCl deprivation is associated with an amyloidogenic shift in the processing of amyloid precursor protein (APP) resulting in an increase of amyloid beta-protein (A beta) secretion. To further investigate this issue we studied the relationship between secretion of APP metabolites (A beta, APPs) and neuronal degeneration. We postulated that the endogenous products of the APP metabolism may modulate neuronal survival by an autocrine loop. Treatment of cerebellar granule cells with various antibodies raised against different epitopes of APPs and A beta oppositely modulates low potassium apoptotic cell death. Antibodies specific for the N-terminal of A beta (4G8, 6E10, R3659) increased neuronal survival by 30% over controls. On the contrary, treatment of cultures undergoing apoptosis with the monoclonal antibody 22C11 directed against the APP N-terminus reduced neuronal survival by 53%, suggesting that endogenous alpha-APPs contribute to neuronal survival. Moreover low KCl culture medium, conditioned by cerebellar granule cells, attenuated the apoptotic process. This anti-apoptotic effect was abolished by removal of APPs from the conditioned medium. Western blotting of APPs removed from the conditioned medium confirmed the presence of alpha-APPs. These data indicate that APP cleavage products oppositely modulate neuronal survival through an autocrine loop and further strengthen an Alzheimer's disease pathogenetic scheme based on altered metabolism of APP. PMID- 10841342 TI - Coding of spatial co-ordinates on neurones of the feline visual association cortex. AB - The receptive field properties of 131 visual neurons lying along the anterior ectosylvian sulcus were studied with extracellular recordings in barbiturate anaesthetized, immobilised, artificially ventilated cats. A stationary visual noise pattern was presented in an area of 24x32 degrees, in which an 8x8 degrees portion moved randomly at a speed of 10 deg/s. A definite best area characterized by maximal firing rate was found, from where the firing rate declined monotonously towards the periphery of the receptive field. The best area locations were distributed in a 40x40 degrees region around the area centralis. These results seem to verify a rather robust capacity in AEV neurones for stimulus localization, in spite of their immense receptive fields. PMID- 10841343 TI - The poly(ADP-ribose) glycohydrolase inhibitor gallotannin blocks oxidative astrocyte death. AB - Poly(ADP-ribose) polymerase (PARP) is now recognized as an important mediator of cell death, but a role for poly(ADP-ribose) glycohydrolase (PARG) in cell death has not previously been described. PARG is the key enzyme degrading ADP-ribose polymers produced by PARP. Here we report effects of the PARG inhibitor gallotannin on oxidative cell death. Pre-incubation of cultured murine astrocytes with as little as 100 nM gallotannin produced significant reductions in H2O2 induced cell death assessed both 24 and 72 h after H2O2 exposure. Gallotannin was more than 10-fold more potent than the PARP inhibitor benzamide in preventing H2O2-induced cell death. These results provide the first evidence that PARG inhibitors could be used to prevent oxidative cell death. PMID- 10841344 TI - Microglia-like cells in rat organ of Corti following aminoglycoside ototoxicity. AB - The repair process of neomycin induced cochlear damage in the postnatal developing rat was investigated in the present study. The results showed that electron dense atypical cells with a cluster of microvilli on their apical surface, resembling early stage of embryonic hair cell, were observed in the former hair cell region. The striking finding was that microglia-like cells appeared and replaced OHCs in the injured auditory sensory epithelium. Using Brdu immunohistochemistry, cell proliferation was found in the area of inner and outer spiral sulcus but not in the hair cells and supporting cells. It is proposed that microglia-like cells play a role in eliminating waste products from the organ of Corti and may participate in direct structural repair. PMID- 10841345 TI - The vitamin C transporter SVCT2 is expressed by astrocytes in culture but not in situ. AB - Ascorbic acid (vitamin C) is known to be selectively accumulated by brain cells through sodium-dependent vitamin C transporters. It is unclear however, whether this uptake occurs in neurons, astrocytes or both. Using Northern analysis we demonstrate that the recently cloned ascorbate transporter isoform SVCT2 is expressed by cultured astrocytes. In contrast, in situ hybridization experiments reveal that SVCT2 mRNA is expressed only in neurons and not in normal astrocytes or astrocytes stimulated by an intrastriatal injection of the neurotoxin quinolinic acid. We conclude that SVCT2 is neuron specific and that the majority of ascorbate storage occurs in neurons. Furthermore, we propose that the observed sodium-dependent ascorbate transport in cultured astrocytes may be due to artificial upregulation of SVCT2 during cell culturing. PMID- 10841346 TI - Detection and localization of BDNF in vestibular nuclei during the postnatal development of the rat. AB - The changes in expression and the subcellular localization of brain-derived neurotrophic factor (BDNF) protein in the rat vestibular nuclear complex (VNC), have been investigated at different postnatal stages. Immunoblotting and ELISA analyses showed a down-regulation of BDNF protein expression in VNC with age. In addition, observations by confocal microscopy revealed that BDNF is mainly located in neuronal somata at postnatal day 8 (P8) and restricted to processes by P15. These results support the idea that BDNF could have different roles in the VNC according to the stage of development The protein could act as a neurotrophic factor in embryonic and early postnatal stages whereas in later developmental stages of the VNC it could be involved in neuronal maturation and regulation of neuronal circuitry. PMID- 10841347 TI - A neural substrate for nonpainful phantom limb phenomena. AB - Activity in the cerebral cortex associated with non-painful phantom limb sensation was studied in 14 upper extremity amputees. In four subjects, repetitive tactile stimulation of the digits or the lower corner of the mouth elicited non-painful phantom sensation in the amputated limb, in the remaining 10 patients no sensation could be evoked. Neuroelectric source imaging revealed significantly elevated activity in SI and posterior parietal cortex, and significantly decreased activity in ipsilateral SII cortex when referred sensations were present. However, nonpainful referred phantom sensations were not associated with a shift of the cortical representation of the mouth into the hand region, as previously suggested. Nonpainful phantom limb experiences seem to have widely distributed neural networks in multiple cortical regions. PMID- 10841348 TI - Microinjection of opiates into the periaqueductal gray matter attenuates neuropathic pain symptoms in rats. AB - We have previously demonstrated that electrical stimulation of the ventral periaqueductal gray matter (PAG) produced analgesia in neuropathic pain in rats. Opioids were also shown to be involved in analgesic effects. This study sought to determine whether opiates microinjected into the ventral PAG produce analgesia. Male Sprague-Dawley rats were chronically implanted with a guide cannula in the PAG under pentobarbital anesthesia and both the tibial and sural nerves were completely cut. Pain sensitivity was postoperatively measured with a von Frey filament and acetone applied to the sensitive area for 1 week. Opioids such as [D Ala2,N-MePhe4,Gly(ol)5]-enkephalin (DAMGO) and [D-Pen ,D-Pen5]-enkephalin (DPDPE) were injected into the PAG. DAMGO, a mu-opioid agonist, and DPDPE, a delta-opioid agonist, were highly effective in reducing neuropathic pain. These effects were reversed by naloxone. These results suggest that the neurons in the ventral PAG are activated by opioids to produce analgesia and that specific opioid receptors are involved in the descending pain inhibition system from the PAG. PMID- 10841350 TI - Differential expression of Bcl-2-related proteins in differentiating NT2 cells. AB - Although the role of Bcl-2-related proteins as regulators of the apoptotic process has been well documented, recent studies suggest that they might also be implicated in neuronal differentiation. We have studied by immunocytochemistry, Western blotting and RT-PCR the expression pattern of Bcl-xL, Bcl-2 and BAX in the in vitro model of neuronal differentiation constituted by retinoic acid (RA) treated NTera-2/D1 (NT2/D1) cells. Whereas BAX level did not change significantly during the RA treatment, Bcl-xL level increased markedly during the first week, before returning to basal level during the second week. Bcl-2 expression, undetectable in undifferentiated cells, increased progressively from the first week. From our results, we suggest that, at least in our model, Bcl-2-related proteins might be involved in neuronal differentiation. PMID- 10841349 TI - Alpha1-adrenoceptors involvement in painful diabetic neuropathy: a role in allodynia. AB - To investigate whether alpha1-adrenoceptors are involved in pain behaviors in streptozotocin (STZ)-induced diabetic rats, we measured the effects of phenylephrine or prazosin on allodynia in the diabetic rats. Phenylephrine aggravated allodynia, while prazosin alleviated allodynia in the diabetic rats. We also measured alpha1-adrenoceptors gene expression or density of [3H]-prazosin binding sites in the dorsal root ganglia (DRG) and spinal cord in painful diabetic rats. Alpha1-adrenoceptors mRNA and density of [3H]prazosin binding sites were increased in the DRG of the diabetic rats, however there were no significant differences in alpha1-adrenoceptors expression in the spinal cord between the control and diabetic rats. These results suggest increased alpha1 adrenoceptors in the DRG may play a role in the pathogenesis of painful diabetic neuropathy. PMID- 10841351 TI - Organization of choroid plexus epithelial and endothelial cell tight junctions and regulation of claudin-1, -2 and -5 expression by protein kinase C. AB - Claudins are components of the tight junctional complex in epithelial and endothelial cells. We characterized the composition of tight junctions in the choroid plexus of the lateral ventricle in the rat brain and tested whether protein kinase C induced changes in their composition. Claudin-1, -2 and -5 were present in the epithelial cells at and near the tight junctions, respectively. In the endothelial cells, claudin-5 was stronger expressed than claudin-1 and -2. Twenty-four hours after the phorbolester injection into the ventricle, claudin-1 immunoreactivity of the epithelial cells was increased and spread to the cytoplasm. The claudin-2 and -5 immunoreactivities were reduced. These findings are consistent with an influence of protein kinase C on the composition of the tight junctions in the choroid plexus. PMID- 10841352 TI - Expression of IL-2 receptor in dorsal root ganglion neurons and peripheral antinociception. AB - Interleukin-2 (IL-2), a T-cell growth factor, could exert an antinociceptive effect. Intraplantar injection of human recombinant IL-2 (rIL-2) at a dose of 10000/20 microl significantly enhanced pain threshold as measured by paw withdrawal latency (PWL) to noxious radiant heat in the rat. Reverse transcription modification of polymerase chain reaction (RT-PCR) analysis showed that IL-2R alpha and beta chains were constitutively expressed in rat dorsal root ganglion (DRG) neurons. Immunohistochemistry further demonstrated that IL-2R was mainly localized to small and medium-sized neurons, which are predominantly responsible for nociceptive transmission. These observations suggest that IL-2/IL 2R system may affect neural activities including nociceptive processing in DRG. PMID- 10841353 TI - Effects of neurotensin receptor antagonists on the firing rate of rat ventral pallidum neurons. AB - Using anaesthetized adult rats, we compared the effects of i.v. injections of neurotensin receptor antagonists (SR48692 and SR142948), haloperidol and clozapine on neuronal firing rate in the two ventral pallidal areas. SR48692 (0.5, 1 and 2 mg/kg, i.v.) induced a dose-dependent inhibition of firing rate in two thirds of neurons in the ventromedial part without any effect in the ventrolateral part. These effects are in keeping with the preferential neurotensin immunoreactivity distribution reported in the ventral pallidum. The classical antipsychotic drug haloperidol (0.5 mg/kg, i.v.) induced an inhibition of neuronal firing rate in both ventral pallidal areas whereas the atypical antipsychotic drug clozapine (20 mg/kg, i.v.), like the neurotensin receptor antagonist SR48692, inhibited cell firing only in the ventromedial part. PMID- 10841354 TI - Electrophysiological evidence for vestibular activation of the guinea pig hippocampus. AB - Vestibular information modulates hippocampal activity for spatial processing and place cell firing. However, evidence of a purely vestibular stimulus modulating hippocampal activity is confounded as most studies use stimuli containing somatosensory and visual components. In the present study, high-frequency electrical stimulation of specific vestibular sensory regions of the right labyrinth in anaesthetized guinea pigs induced an evoked field potential in the hippocampal formation bilaterally with a latency of about 40 ms following stimulation onset. Field potentials localized in the hippocampal formation occurred with stimulus current parameters that were too small to produce eye movements. This provides direct electrophysiological evidence of vestibular input to the hippocampus. PMID- 10841355 TI - SNC 80, a delta-opioid agonist, elicits phase advances in hamster circadian activity rhythms. AB - Non-photic stimuli administered to hamsters during the subjective day can cause phase advances in circadian wheel running activity. It is believed that afferent projections from the intergeniculate leaflet of the thalamus to circadian pacemaker cells within the suprachiasmatic nucleus mediate the phase shifting effects of some non-photic stimuli. In hamsters, many of the intergeniculate leaflet afferents contain enkephalin, yet the role of opioids in producing non photic phase shifts in hamsters has not been reported. In the present study, we show that SNC 80, an agonist for the delta opioid receptor subtype, will phase advance hamster wheel running activity rhythms when administered late in the subjective day. These results indicate that opioids may be involved in modulating the circadian pacemaker in hamsters. PMID- 10841356 TI - Axonal transport of neuronal calcium sensor-1 and phosphatidylinositol 4-kinase beta in the adult rat sciatic nerve. AB - Neuronal calcium sensor-1 (NCS-1) and its putative substrate phosphatidylinositol 4-kinase beta (PtdIns 4-kinase beta) both indirectly regulate synaptic vesicle exocytosis and are located in DRG neurites. In this study we have tested whether NCS-1 and PtdIns 4-kinase beta are transported in axons using the analysis of double ligation approach in the adult rat sciatic nerve. We show that NCS-1 accumulates on both the distal and proximal side of the nerve ligation indicating that this protein undergoes bidirectional transport in axons. In contrast, PtdIns 4-kinase beta accumulated on the distal side which suggests that it undergoes retrograde axonal transport and unlike NCS-1 was also present in non-neuronal cells. PMID- 10841357 TI - Normal discrimination performance accompanied by priming deficits in monkeys with V4 or TEO lesions. AB - Primate visual areas V4 and TEO are important for many aspects of visual perception and ablation of these areas leads to a wide range of deficits in visual discrimination, attention to less salient items, recognition of visually transformed objects, visual grouping and in visual memory. All these studies demonstrate that monkeys with V4 or TEO lesions have higher perceptual thresholds or are slower or less accurate than normal monkeys on a particular visual task. Here we show that when monkeys with V4 or TEO lesions perform a simple discrimination task on which they are unimpaired, they perform the task differently from normal monkeys. We examined visual priming in a feature detection task and discovered that it is diminished by lesions of TEO and abolished by lesions of V4. The results support the hypothesis, based on a recent demonstration of visuotopic priming in humans, that areas V4 and TEO are indispensable for normal visual form priming. PMID- 10841358 TI - Increased distractibility in closed head injury as revealed by event-related potentials. AB - The present study demonstrates that event-related potentials (ERPs) may be used to reveal increased distractibility as a physiologically measurable condition after chronic closed head injury (CHI). ERPs were recorded from 17 chronic CHI subjects and from 17 healthy age-matched controls. Auditory stimuli consisted of variants of vowel /o/ (standards) occasionally replaced by an /e/ vowel (deviant). Subjects were instructed to ignore auditory stimuli while watching a silent movie. In the constant-standard condition, the vowel /o/ served as the standard and vowel /e/ as the deviant. In the roving-standard condition, four variants of the vowel /o/ were randomly used as standards in the same stimulus block. None of the stimuli were prototypes in the subjects' mother tongues. Deviant stimuli elicited significant MMNs in both groups in both conditions, which were significantly smaller in the roving-standard than in the constant standard condition. CHI victims showed significantly larger P3a amplitudes than controls in both conditions, apparently reflecting their enhanced involuntary sifting of attention and thus their increased distractibility. PMID- 10841359 TI - Auditory cortex activation associated with octave illusion. AB - Most right-handers perceive an octave illusion when they are presented with a 400 Hz tone to one ear and with a 800 Hz tone to the other ear simultaneously, and when the tones continuously reverse between the ears: instead of the correct sound sequence, the subjects typically report a high tone in the right ear alternating with a low tone in the left. To study the neural basis of the illusion, we recorded neuromagnetic responses to binaural 400 and 800 Hz tones in different combinations. In the auditory cortex of each hemisphere, the 100 ms response (N100m) was stronger to pairs where the 800 Hz tone was presented to the contralateral ear and the 400 Hz tone to the ipsilateral ear than vice versa. The sustained fields tended to behave in an opposite manner. We suggest that the perceived locations of the sounds in the octave illusion follow the N100m lateralization, and the percept is contributed by streaming by the ear. PMID- 10841360 TI - NGF-induced motoneuron cell death depends on the genetic background and motoneuron sub-type. AB - Nerve growth factor (NGF) promotes the survival of several neuronal populations, but recently it has also been shown to induce neuronal cell death. Here we report the effects of NGF on lesioned motoneurons. We have analyzed facial and sciatic motoneurons in newborn and adult BALB/c and C57BL/6 mice, in addition to mice deficient in the low-affinity p75 receptor for the neurotrophins (p75NTR). NGF application did not alter survival of lesioned facial motoneurons in any of the strains examined independent of the age of the animals. Only in the adult C57BL/6 mouse strain where the sciatic nerve had been crushed prior to factor application did NGF induce cell death of axotomized sciatic motoneurons. Our results illustrate the importance of the genetic background and the motoneuron sub-type in studies related to cell death and survival of motoneurons in relation to NGF and p75NTR. PMID- 10841361 TI - GFP fluorescence reports Period 1 circadian gene regulation in the mammalian biological clock. AB - Endogenous cyclic activation of a specific set of genes, including Period 1 (Per1), drive circadian rhythms in the suprachiasmatic nucleus (SCN), a biological clock nucleus of the brain. We have produced transgenic mice in which a degradable form of recombinant jellyfish green fluorescent protein (GFP) is driven by the mouse Period 1 (mPer1) gene promoter. GFP protein is expressed in the circadian neural structures of the retina and SCN. Fluorescent signals are resolved at the level of individual neurons. mPer1-driven GFP fluorescence intensity reports light-induction and circadian rhythmicity in SCN neurons. This circadian reporter transgene captures the gene expression dynamics of living biological clock neurons and ensembles, providing a novel view of this brain function. PMID- 10841362 TI - Leu-enkephalin induced by IL-2 administration mediates analgesic effect of IL-2. AB - Interleukin-2 (IL-2) was found to have an analgesic effect in both central and peripheral nervous systems. This effect is related to opioid receptors and mediated mainly by IL-2 directly binding to opioid receptors. Using radioimmunoassay, the content of Leu-enkephalin (LEK) in some nuclei were measured at intervals after the injection of IL-2 into the lateral ventricle of rats. Levels of LEK increased in both paraventricular hypothalamic nucleus (PVN) and locus ceruleus (LOC) after IL-2 administration, suggesting that the analgesic effect of IL-2 is also related to the change of LEK in PVN and LOC induced by IL 2 administration. PMID- 10841363 TI - fMRI shows multiple somatotopic digit representations in human primary somatosensory cortex. AB - Using electrical finger nerve stimulation in normal human subjects, fMRI detected separate representations for all 5 fingers in the primary somatosensory cortex. Responses were located in the posterior wall of the deep central sulcus (most likely corresponding to Brodmann Area (BA) 3b), and the anterior (BA 1) or posterior crown of the postcentral gyrus (BA 2) with rare activations in BA 3a and 4. In BA 3b we found a regular somatotopic mediolateral digit arrangement for fingers 5 to 1 with a mean Euclidean distance of 16 mm between fingers 1 and 5. In contrast BA 1/2 showed a greater number of adjacent activation foci with significantly more overlap and partly even reversed ordering of neighbouring fingers. PMID- 10841364 TI - Prenatal viral infection causes alterations in nNOS expression in developing mouse brains. AB - Epidemiological evidence points to prenatal viral infection being responsible for some forms of schizophrenia and autism. We hypothesized that prenatal human influenza viral infection in day 9 pregnant mice may cause changes in the levels of neuronal nitric oxide synthase (nNOS), an important molecule involved in synaptogenesis and excitotoxicity, in neonatal brains. Brains from 35- and 56-day old mice were prepared for SDS-gel electrophoresis and Western blotting using polyclonal anti nNOS antibody. Quantification of nNOS showed time and region dependent changes in the levels of nNOS protein. Mean rostral brain area value from prenatally infected animals showed a significant (p=0.067) increase of 147% in nNOS levels at 35 days postnatally, with an eventual 29% decrease on day 56. Middle and caudal brain areas showed reductions in nNOS in experimental mice at 35 and 56 days, with a significant 27% decrease in nNOS in the middle segment of day 56 brains (p=0.016). Significant interactions were found between group membership and brain area (Wilks lambda=0.440, F(2.9)=5.72, p=0.025); there was also a significant interaction between brain area, group and age (Wilks lambda=0.437, F(2.9)=5.79, p=0.024). These results provide further support for the notion that prenatal viral infection affects brain development adversely via the pathological involvement of nNOS expression. PMID- 10841365 TI - Expression of connexin36 mRNA in adult rodent brain. AB - A new member of the connexin gene family, named Connexin36 (Cx36) has, recently, been identified in rodents and shown to be preferentially, if not exclusively, expressed in neurones of the adult CNS. In this study we present a detailed in situ hybridization analysis of the expression pattern of mouse Connexin 36 (mCx36) mRNA in the adult mouse brain, with particular regards to the correlation of mCx36 expression to specific neuronal cell classes and systems. We found that mCx36 was strongly and widely expressed in the brain, including areas where the presence of gap junctions was never detected before. Quantitative analysis of the hybridization signal indicated varying levels of expression in different areas. In particular mCx36 was highly expressed in the neurones at different levels of the motor pathway, the olfactory pathway, the hippocampus, and areas related to the generation of respiratory rhythm. On the contrary, mCx36 was more heterogeneously expressed in nuclei of the sensory pathways. These findings show that mCx36 is the first connexin specifically expressed in neuronal cells in the adult rodent brain. The profiles of expression clearly indicate that mCx36 might play specific roles within different neuronal systems. PMID- 10841366 TI - Mismatch negativity to inclusions and omissions of stimulus features. AB - Two experiments were run to determine the effects of addition or removal of a stimulus feature on mismatch negativity (MMN). In the first experiment, a deviant stimulus was constructed by adding a click to a white noise standard stimulus. In the second experiment, the deviant was constructed by subtracting the click from the standard. In different conditions, the intensity of the click was varied. When the deviant was constructed by the addition of a click, a significant MMN was evident in those conditions in which click-to-noise ratio exceeded 1.0. When the deviant was constructed by the subtraction of the click, a significant negativity was found only when the click-to-noise ratio was very large. However, this negativity was accompanied by only a small polarity inversion at the mastoid. The MMN is thus best elicited when the deviant stimulus contains a new afferent element not present in the standard stimulus. PMID- 10841367 TI - Amplitude envelope correlation detects coupling among incoherent brain signals. AB - Cognitive processing involves gamma-activation over broad cortical regions. Phase coupling of these activities has rarely been reported for areas far apart. Other forms of coupling are generally not detected by conventional measures. Here, we use amplitude envelope correlation (AEC), which can detect signal coupling without phase coherence, even among different frequencies. We apply it to subdural recordings from humans performing a visual delayed match-to-sample task and systematically compare it with spectral amplitude and coherence. The different measures often show divergent results. In particular, AEC reveals y coupling completely missed by coherence. We argue that coherence and AEC are adapted to different cortical mechanisms of short- and long-range interactions, respectively. PMID- 10841368 TI - Cortical and subcortical visual event-related potentials to oddball stimuli in rabbits. AB - Event-related potentials (ERPs) to changes in the visual environment were recorded in rabbits. In the oddball condition, infrequently presented (deviant) stimuli occurred in a series of frequently presented (standard) stimuli. In the deviant-alone condition, standards were omitted. ERPs to oddball-deviants differed from those to standards in all recording sites (cerebellar cortex, visual cortex, dentate gyrus). No corresponding differences were found between ERPs to deviants in the oddball condition and those in the deviant-alone condition. However, because ERPs to deviants in the deviant-alone condition and those to standards did not differ either, ERPs to stimulus changes in the oddball condition seemed to be dependent on the presence of standards, thus representing an analogue to mismatch negativity (MMN) in humans. PMID- 10841369 TI - Contralateral neglect induced by right posterior parietal rTMS in healthy subjects. AB - We applied repetitive transcranial magnetic stimulation (rTMS) in order to induce interference on visuo-spatial perception in 11 healthy subjects. Subjects performed a visuo-spatial task requiring judgements about the symmetry of prebisected lines. Visual stimuli consisted of symmetrically or asymmetrically transected lines, tachystoscopically presented for 50 ms on a computer-monitor. Performance was examined in basal condition and during rTMS trains of 10 stimuli at 25 Hz, delivered through a focal coil over right or left posterior parietal cortex (P5 and P6 sites) and triggered synchronously with visual stimulus. Randomly intermixed sham rTMS trains were employed to control for non-specific effects. Right parietal rTMS induced a significant rightward bias in symmetry judgements as compared with basal and sham rTMS conditions. No differences emerged between other conditions. PMID- 10841370 TI - A thalamo-prefrontal system for representation in executive response choice. AB - This study demonstrates the neural system potentially involved in the representation of, and choice between, stimulus classifications in an ambiguous, novel, decision-making task. This difficult choice behaviour is taken as an example of a basic executive processing task. Subjects heard sounds that were consonant-vowel combinations that had been distorted and were required to categorize each stimulus as speech-like or not-speech-like. Cerebral activity was measured with positron emission tomography. A neural system (thalamic and medial prefrontal cortical regions) was demonstrated; there was greater activity involved in assigning the sound to the larger class of not-speech-like sounds than to the more restricted category of speech-like sounds. We interpret this activity as reflecting process and representation in a simple central executive task. PMID- 10841371 TI - Selective up-regulation of I kappaB-alpha in ischemic penumbra following focal cerebral ischemia. AB - Activation of transcription factor NF-kappaB plays a critical role in immune, inflammatory and cell death responses. In resting cells, NF-kappaB is sequestrated in the cytoplasm in an inactive form through its association with inhibitory proteins, I kappaB (e.g.I kappaB-alpha). In response to cell activation, I kappaB is degraded causing release of active NF-kappaB. Active NF kappaB translocates into the nucleus leading to activation of transcription that may have a profound effect on cell survival, including that after ischemic stroke. Here, using Western blot analysis, we show that immunoreactivity to the major subunit of NF-kappaB, p65, as well as to the inhibitory subunit I kappaB alpha is equally markedly decreased in the ischemic core after transient middle cerebral and common carotid artery occlusion in rats. In contrast, penumbral regions display no change in p65, and significant increase in I kappaB-alpha immunoreactivity, as compared to non-ischemic areas. In these penumbral regions with elevated I kappaB-alpha immunoreactivity, we find reduced cytosolic and increased nuclear I kappaB-alpha staining of neurons, as determined by immunohistochemistry. Altogether, these results suggest that an altered ratio between activating and inhibitory NF-kappaB pathways mediated through I kappaB alpha may play an important role in survival of the ischemic penumbra. PMID- 10841372 TI - Sound localization in the human brain: neuromagnetic observations. AB - Sound location processing in the human auditory cortex was studied with magnetoencephalography (MEG) by producing spatial stimuli using a modern stimulus generation methodology utilizing head-related transfer functions (HRTFs). The stimulus set comprised wideband noise bursts filtered through HRTFs in order to produce natural spatial sounds. Neuromagnetic responses for stimuli representing eight equally spaced sound source directions in the azimuthal plane were measured from 10 subjects. The most prominent response, the cortically generated N1m, was investigated above the left and right hemisphere. We found, firstly, that the HRTF-based stimuli presented from different directions elicited contralaterally prominent N1m responses. Secondly, we found that cortical activity reflecting the processing of spatial sound stimuli was more pronounced in the right than in the left hemisphere. PMID- 10841373 TI - Prenatal exposure to SKF-38393 alters the response to light of adult rats. AB - The current study examined the consequences of prenatal SKF-38393 exposure on the cellular response in the adult suprachiasmatic nuclei to light. Pregnant rats were injected with the dopamine agonist SKF-38393 or vehicle daily from gestational day 15 to 21. Adult offspring received a light pulse (1 min/2 lux) 4 or 8 h after lights off (ZT16 or ZT20 where ZT=zeitgeber time). Brains were processed for c-FOS-like immunoreactivity in the SCN. At ZT20 the number of cells expressing c-FOS protein after a light pulse was the same in both groups. At ZT16 the number of cells in the SCN of SKF-38393-exposed animals was 58% lower than the vehicle-treated group. The data suggest that prenatal SKF-38393 treatment may have long-term consequences for SCN function. PMID- 10841374 TI - Inhibition of N-glycan processing alters axonal transport of synaptic glycoproteins in vivo. AB - Synaptic glycoproteins are synthesized and glycosylated in the neuronal cell body, and conveyed to terminals by fast axonal transport. We used the alpha mannosidase inhibitor, 2-deoxymannojirimycin (dMan), to investigate the effects of disrupting N-glycan processing on the axonal trafficking of proteins in vivo. dMan significantly reduced rapid axonal transport in retinal ganglion cells to about 34% of control values 4h after metabolic labeling; at 8 h post-labeling the inhibition was reversed. 2-D gel analysis showed that dMan completely inhibited the arrival of radiolabeled L1 and NCAM at axon terminals, and resulted in the appearance of two novel proteins of 230 kDa and 155 kDa. Our results show that disruption of the N-glycosylation pathway has an immediate inhibitory effect on total axonal transport and longer lasting effects on the trafficking of specific glycoproteins to axon terminals in vivo. PMID- 10841375 TI - Relative contributions from neuronal and endothelial nitric oxide synthases to regional cerebral blood flow changes during forebrain ischemia in rats. AB - The principal aim of this study was to examine the relative contributions from the neuronal and endothelial isoforms of nitric oxide synthase (nNOS and eNOS, respectively) in their capacity to modulate intra-ischemic cerebral blood flow (CBF) changes, in the ischemically vulnerable hippocampus and striatum. CBF changes were monitored, using laser-Doppler flowmetry, in rats subjected to 30 min of forebrain ischemia (right common carotid occlusion+hemorrhagic hypotension). Rats were pretreated with a selective nNOS inhibitor (ARR 17477), a NOS inhibitor that blocks both eNOS and nNOS (N(G)-nitro-L-arginine; L-NNA), or saline (control). In initial experiments, where ischemic MABP was targeted to exactly 30 mmHg, NOS inhibition reduced intra-ischemic cortical CBF from the control level of approximately 20% of baseline to 3% (L-NNA) or 6% (ARR 17477) of baseline. The statistically similar effects of the two NOS inhibitors confirmed that nNOS is the predominant NO source supporting intra-ischemic vasodilation in the cortex. In subsequent experiments, CBF was measured in the right hippocampus, and striatum, as well as the cortex, and, to reduce data variability, blood withdrawal was adjusted to achieve an intra-ischemic cortical CBF of 20% (controls) or 5% (NOS inhibited rats) of baseline. In those groups, mean ischemic MABP levels ranged from 28 to 32 mmHg. In controls, intra-ischemic CBF fell to 20%, 45%, and 47% of baseline in the cortex, hippocampus, and striatum, respectively. With nNOS inhibition, intra-ischemic CBF was further reduced to 5%, 15%, and 18% of baseline, respectively. However, with combined eNOS/nNOS inhibition, the CBF values were 5%, 37%, and 21%, respectively. These results suggest that the nNOS contribution to intra-ischemic vasodilation in vulnerable regions is substantially greater than eNOS. The significantly higher intra ischemic CBF level in the hippocampus in combined eNOS/nNOS vs nNOS-inhibited rats may relate, in contrast to other regions, to a low eNOS influence on vascular function in that structure and CBF redistribution to the hippocampus when eNOS activity is blocked globally. PMID- 10841376 TI - Hypoxia influences enkephalin release in rats. AB - To investigate hypoxia effects on leucine-enkephalin (L-ENK) levels of median eminence (ME) of hypothalamus in rats, and the possible glucocorticoid involvement in modulation. Hypoxia was stimulated in a hypobaric chamber. L-ENK levels were measured by specific radioimmunoassay during acute, and a comparison of L-ENK alteration was taken for bilateral adrenalectomy (ADX) with or without a replacement of dexamethasone (DEX). Acute hypoxia 10.8% O2 for 30 min and 2 h markedly enhanced L-ENK levels of ME from 23.99+/-7.25 in control to 51.26+/ 16.96 (P<0.01) and 53.29+/-26.10 ng/mg protein (p<0.01), and acute hypoxia at 8.2% O2 significantly increased L-ENK of ME to 36.76+/-15.25 (p<0.05) and 32.09+/ 3.58 ng/mg protein (p<0.05). The increased L-ENK was returned to normoxic level when hypoxia (10.8% O2) exposure lasted for 24 h. After ADX, 10.8% O2 hypoxia induced a sharp decline of L-ENK in the ME, but this decline was completely reversed by treatment with DEX (500 microg/rat, i.p.). Acute hypoxia increases L ENK level of hypothalamic ME in rats, which may present a reduced and/or inhibited release of L-ENK, acting through a fast negative feedback mechanism of acute hypoxia activated high circulating glucocorticoid level. PMID- 10841377 TI - Central processing of human ocular torsion analyzed by galvanic vestibular stimulation. AB - We examined the dynamics of human ocular torsion (OT) responses to sinusoidal galvanic vestibular stimulation (GVS) (0.005-1.67 Hz). The tonic OT showed a lowpass characteristic with a time constant of 1.74 s and a gain of 0.93 deg/mA. In two subjects, nystagmus dominated the observable OT pattern at frequencies <0.1 Hz. The nystagmus slow phases showed an exponential trajectory with a time constant of 1.49 s. The dynamics of both tonic OT and torsional nystagmus in our study were similar to the dynamics of OT induced by rotation and linear acceleration found in the literature. We propose a model for the central processing of torsional eye movements that is based on a common neural integration of semicircular canal (SC) and utricular inputs as well as nystagmus bursts. The sensitivity of all vestibular afferents to GVS was derived to be 0.76 spikes/s/mA. SC effects on OT are at least 3.5 times higher than utricular effects. PMID- 10841378 TI - Plasticity revealed by transcranial magnetic stimulation of early visual cortex. AB - We applied single-pulse transcranial magnetic stimulation (TMS) to the occipital pole of healthy subjects while they performed a forced-choice visual letter identification task. We found three separate periods when TMS suppressed performance; the first period is best explained by TMS-induced blinking whereas the last two periods are best explained by TMS-induced disruption of letter processing in the early visual cortex. Unexpectedly, we also found that TMS induced suppression progressively disappeared during three weeks of repeated TMS experiments. However, it was only suppression during the last two periods that disappeared; suppression during the first period remained undiminished. When subjects were then presented with dimmer letters, suppression reappeared. The most likely explanation is a practice-induced increase in neuronal activity in the early visual cortex. PMID- 10841379 TI - Retinoic acid delays transcription of human retinal pigment neuroepithelium marker genes in ARPE-19 cells. AB - The effect of retinoic acid on the differentiation of a human retinal pigment epithelium-derived cell line ARPE-19 was studied. Differentiation of ARPE-19 cells is delayed by retinoic acid. The minimum all-trans-retinoic acid concentration needed for delay of ARPE-19 differentiation is 1 microM. A delay of differentiation was also observed using 1 microM 9-cis or 13-cis-retinoic acid. Differentiation at the molecular level was studied by analyzing transcription of two RPE-marker genes, RPE65 and peropsin. In the presence of 1 microM retinoic acid the onset of transcription of both genes was delayed by 2-3 weeks. We conclude that all-trans-, 9-cis-, and 13-cis-retinoic acid delay differentiation of ARPE-19 cells into cells that phenotypically resemble cells from the human retinal pigment epithelium. PMID- 10841380 TI - Functional brain mapping of the relaxation response and meditation. AB - Meditation is a conscious mental process that induces a set of integrated physiologic changes termed the relaxation response. Functional magnetic resonance imaging (fMRI) was used to identify and characterize the brain regions that are active during a simple form of meditation. Significant (p<10(-7)) signal increases were observed in the group-averaged data in the dorsolateral prefrontal and parietal cortices, hippocampus/parahippocampus, temporal lobe, pregenual anterior cingulate cortex, striatum, and pre- and post-central gyri during meditation. Global fMRI signal decreases were also noted, although these were probably secondary to cardiorespiratory changes that often accompany meditation. The results indicate that the practice of meditation activates neural structures involved in attention and control of the autonomic nervous system. PMID- 10841381 TI - Neuropeptide Y applied in vitro can block the phase shifts induced by light in vivo. AB - The mammalian suprachiasmatic nuclei (SCN) can be synchronized by light, with direct glutamatergic input from the retina. Input to the SCN from the intergeniculate leaflet contains neuropeptide Y (NPY) and can modulate photic responses. NPY can reduce the phase-resetting effect of light or glutamate. We investigated the effect of NPY applied in vitro on light-induced phase shifts of the SCN neural activity rhythm. Light pulses delivered in vivo induced phase shifts in brain slice preparations similar to those as measured by behavioral activity rhythms. NPY applied after the light pulse blocked the phase shifts during both the early and late subjective night. NPY applied 30 min after the light pulse could block the phase delay induced by light. Our results show that NPY can inhibit photic resetting of the clock during the subjective night. The time course of this inhibitory effect suggests a mechanism downstream of the glutamate receptor. PMID- 10841382 TI - High frequency stimulation of the STN influences the activity of dopamine neurons in the rat. AB - The effect of high frequency stimulation (HFS) of the subthalamic nucleus (STN) on the spontaneous activity of substantia nigra pars compacta (SNc) dopaminergic neurons was investigated in normal rats and in rats with globus pallidus (GP) lesions. In normal rats, the spontaneous activity of SNc neurons did not significantly differ from that of rats with GP lesions (4.2+/-2.2 versus 4.4+/ 2.6 spikes/s). STN-HFS induced an increase of firing rate in the majority of tested cells in normal (76%) and GP-lesioned rats (73%) with an after-effect of 34.4+/-3.4 and 33.2+/-3.1 s, respectively. These results demonstrate that STN-HFS influences the activity of the SNc dopaminergic neurons by increasing their firing rate and that this increase of activity is independent of the globus pallidus. PMID- 10841383 TI - Candidacy for board certification in radiological/medical physics should be restricted to graduates of accredited training programs. PMID- 10841384 TI - Calculation of effective dose. AB - The concept of "effective dose" was introduced in 1975 to provide a mechanism for assessing the radiation detriment from partial body irradiations in terms of data derived from whole body irradiations. The effective dose is the mean absorbed dose from a uniform whole-body irradiation that results in the same total radiation detriment as from the nonuniform, partial-body irradiation in question. The effective dose is calculated as the weighted average of the mean absorbed dose to the various body organs and tissues, where the weighting factor is the radiation detriment for a given organ (from a whole-body irradiation) as a fraction of the total radiation detriment. In this review, effective dose equivalent and effective dose, as established by the International Commission on Radiological Protection in 1977 and 1990, respectively, are defined and various methods of calculating these quantities are presented for radionuclides, radiography, fluoroscopy, computed tomography and mammography. In order to calculate either quantity, it is first necessary to estimate the radiation dose to individual organs. One common method of determining organ doses is through Monte Carlo simulations of photon interactions within a simplified mathematical model of the human body. Several groups have performed these calculations and published their results in the form of data tables of organ dose per unit activity or exposure. These data tables are specified according to particular examination parameters, such as radiopharmaceutical, x-ray projection, x-ray beam energy spectra or patient size. Sources of these organ dose conversion coefficients are presented and differences between them are examined. The estimates of effective dose equivalent or effective dose calculated using these data, although not intended to describe the dose to an individual, can be used as a relative measure of stochastic radiation detriment. The calculated values, in units of sievert (or rem), indicate the amount of whole-body irradiation that would yield the equivalent radiation detriment as the exam in question. In this manner, the detriment associated with partial or organ-specific irradiations, as are common in diagnostic radiology, can be assessed. PMID- 10841385 TI - Effective doses to patients undergoing thoracic computed tomography examinations. AB - The purpose of this study was to investigate how x-ray technique factors and effective doses vary with patient size in chest CT examinations. Technique factors (kVp, mAs, section thickness, and number of sections) were recorded for 44 patients who underwent a routine chest CT examination. Patient weights were recorded together with dimensions and mean Hounsfield unit values obtained from representative axial CT images. The total mass of directly irradiated patient was modeled as a cylinder of water to permit the computation of the mean patient dose and total energy imparted for each chest CT examination. Computed values of energy imparted during the chest CT examination were converted into effective doses taking into account the patient weight. Patient weights ranged from 4.5 to 127 kg, and half the patients in this study were children under 18 years of age. All scans were performed at 120 kVp with a 1 s scan time. The selected tube current showed no correlation with patient weight (r2=0.06), indicating that chest CT examination protocols do not take into account for the size of the patient. Energy imparted increased with increasing patient weight, with values of energy imparted for 10 and 70 kg patients being 85 and 310 mJ, respectively. The effective dose showed an inverse correlation with increasing patient weight, however, with values of effective dose for 10 and 70 kg patients being 9.6 and 5.4 mSv, respectively. Current CT technique factors (kVp/mAs) used to perform chest CT examinations result in relatively high patient doses, which could be reduced by adjusting technique factors based on patient size. PMID- 10841387 TI - Using light sensitometry to evaluate mammography film performance. AB - The performance of commercially available light sensitometers was compared with two other methods of x-ray sensitometry to determine whether commercially available sensitometers are viable for evaluating clinical performance of mammography film. X-ray sensitometry was performed using mammography screens that were modified to accommodate a graded optical step tablet (screen sensitometry). Finally, a means for performing intensity-scale x-ray sensitometry was configured (inverse-square sensitometry). Clinical mammography x-ray exposure conditions were used and film processing quality was closely monitored during the study. Statistical results for chi-square probabilities on the resulting contrast curves yielded good agreement for most of the configurations investigated. Comparison of film gradient versus optical density curves showed good agreement for maximum contrast values and the corresponding optical density for maximum contrast for three of the four screen-film combinations used when comparing light sensitometry to screen sensitometry. A similar comparison of light sensitometers to inverse square sensitometry showed good agreement for maximum contrast, but less agreement for the corresponding optical density of maximum contrast. Based on these results, the authors concluded that commercially available sensitometers could be used to estimate clinical film performance for the screen-film systems tested. In particular they can be used to determine the range of optical densities that provide optimal film contrast. PMID- 10841386 TI - An edge spread technique for measurement of the scatter-to-primary ratio in mammography. AB - An experimental measurement technique that directly measures the magnitude and spatial distribution of scatter in relation to primary radiation is presented in this work. The technique involves the acquisition of magnified edge spread function (ESF) images with and without scattering material present. The ESFs are normalized and subtracted to yield scatter-to-primary ratios (SPRs), along with the spatial distributions of scatter and primary radiation. Mammography is used as the modality to demonstrate the ESF method, which is applicable to all radiographic environments. Sets of three images were acquired with a modified clinical mammography system employing a flat panel detector for 2, 4, 6, and 8 cm thick breast tissue equivalent material phantoms composed of 0%, 43%, and 100% glandular tissue at four different kV settings. Beam stop measurements of scatter were used to validate the ESF methodology. There was good agreement of the mean SPRs between the beam stop and ESF methods. There was good precision in the ESF determined SPRs with a coefficient of variation on the order of 5%. SPRs ranged from 0.2 to 2.0 and were effectively independent of energy for clinically realistic kVps. The measured SPRs for 2, 4, and 6 cm 0% glandular phantoms imaged at 28 kV were 0.21+/-0.01, 0.39+/-0.01, and 0.57+/-0.02, respectively. The measured SPRs for 2, 4, and 6 cm 43% glandular phantoms imaged at 28 kV were 0.20+/-0.01, 0.35+/-0.02, and 0.53+/-0.02, respectively. The measured SPRs for 2, 4, and 6 cm 100% glandular phantoms imaged at 28 kV were 0.22+/-0.02, 0.42+/ 0.03, and 0.88+/-0.08, respectively. PMID- 10841388 TI - High temporal resolution for multislice helical computed tomography. AB - Multislice helical computed tomography (CT) substantially reduces scanning time. However, the temporal resolution of individual images is still insufficient for imaging rapidly moving organs such as the heart and adjacent pulmonary vessels. It may, in some cases, be worse than with current single-slice helical CT. The purpose of this study is to describe a novel image reconstruction algorithm to improve temporal resolution in multislice helical CT, and to evaluate its performance against existing algorithms. The proposed image reconstruction algorithm uses helical interpolation followed by data weighting based on the acquisition time. The temporal resolution, the longitudinal (z-axis) spatial resolution, the image noise, and the in-plane image artifacts created by a moving phantom were compared with those from the basic multislice helical reconstruction (helical filter interpolation, HFI) algorithm and the basic single-slice helical reconstruction algorithm (180 degrees linear interpolation, 180LI) using computer simulations. Computer simulation results were verified with CT examinations of the heart and lung vasculature using a 0.5 second multislice scanner. The temporal resolution of HFI algorithm varies from 0.28 and 0.86 s, depending on helical pitch. The proposed method improves the resolution to a constant value of 0.29 s, independent of pitch, allowing moving objects to be imaged with reduced blurring or motion artifacts. The spatial (z) resolution was slightly worse than with the HFI algorithm; the image noise was worse than with the HFI algorithm but was comparable to axial (step-and-shoot) CT. The proposed method provided sharp images of the moving objects, portraying the anatomy accurately. The proposed algorithm for multislice helical CT allowed us to obtain CT images with high temporal resolution. It may improve the image quality of clinical cardiac, lung, and vascular CT imaging. PMID- 10841389 TI - A z gain nonuniformity correction for multislice volumetric CT scanners. AB - This paper presents a calibration and correction method for detector cell gain variations. A key functionality of current CT scanners is to offer variable slice thickness to the user. To provide this capability in multislice volumetric scanners, while minimizing costs, it is necessary to combine the signals of several detector cells in z, when the desired slice thickness is larger than the minimum provided by a single cell. These combined signals are then pre-amplified, digitized, and transmitted to the system for further processing. The process of combining the output of several detector cells with nonuniform gains can introduce numerical errors when the impinging x-ray signal presents a variation along z over the range of combined cells. These numerical errors, which by nature are scan dependent, can lead to artifacts in the reconstructed images, particularly when the numerical errors vary from channel-to-channel (as the filtered-backprojection filter includes a high-pass filtering along the channel direction, within a given slice). A projection data correction algorithm has been developed to subtract the associated numerical errors. It relies on the ability of calibrating the individual cell gains. For effectiveness and data flow reasons, the algorithm works on a single slice basis, without slice-to-slice exchange of information. An initial error vector is calculated by applying a high pass filter to the projection data. The essence of the algorithm is to correlate that initial error vector, with a calibration vector obtained by applying the same high-pass filter to various z combinations of the cell gains (each combination representing a basis function for a z expansion). The solution of the least-square problem, obtained via singular value decomposition, gives the coefficients of a polynomial expansion of the signal z slope and curvature. From this information, and given the cell gains, the final error vector is calculated and subtracted from the projection data. PMID- 10841390 TI - A localization algorithm and error analysis for stereo x-ray image guidance. AB - Stereo x-ray radiography attracts increasing attention in major clinical applications. The purpose of this paper is to analyze the 3D localization error for breast biopsy procedures and provide guidelines for improving its accuracy. Our prototype is a CCD based digital stereo x-ray imaging system. The mathematical model consists of two x-ray sources and one stationary detector plane. A closed form least-squares solution is derived for 3D localization of feature points, particularly a biopsy needle tip, from a pair of 2D digital radiographs. Based on the least-squares formula and its first order approximation, the 3D localization error is analyzed in terms of object location, measurement error, separation between the two x-ray sources, and distance from the source to the detector. The stereo imaging and error estimation formulas are numerically simulated and experimentally validated. The data are in agreement with theoretical prediction. These results can be used for the purpose of system design and protocol optimization. PMID- 10841391 TI - Anthropomorphic versus geometric chest phantoms: a comparison of scatter properties. AB - Previously, we have used an anthropomorphic chest phantom to study scatter reduction in digital chest radiography. Image metrics, such as scatter fractions, contrast, noise, and resolution, are not easily measured due to the anatomical structure in the phantom. A geometric chest phantom, recently developed for quality control purposes, offers the possibility of being used to calculate image quality measurements. Here, we compare the scatter properties of the two phantoms to determine if the geometric phantom can be used in our studies of scatter compensation techniques. A calibrated photostimulable phosphor system was used to acquire images of the two phantoms. An array of beam stops was placed in front of each phantom to calculate scatter fractions. Each phantom had approximately 2 in. of polystyrene material added to the posterior to increase scatter fractions to those normally seen in patients. Exposure parameters were 300 mA for 0.009 sec with a source to image distance of 100 cm. Energies were varied from 60 to 130 kVp. Scatter fractions were determined for different areas of anatomy for each energy and each phantom. For all energies examined, the two phantoms compare well for scatter fractions in each of six regions. For example, at 95 kVp, the geometric phantom had average scatter fractions of 0.72 and 0.88 in the lung and mediastinum regions, respectively. These values were 0.74 and 0.90 for the anatomic phantom. For comparison, measurements of scatter fractions in patients at these values have been reported as 0.65 and 0.90 for the lung and mediastinum regions. The geometric phantom is an excellent tool which can be used in place of the anthropomorphic phantom for studies involving scatter compensation. In addition to having a gray level histogram typical of a human chest, this phantom has uniform regions where image quality measurements can be calculated. PMID- 10841392 TI - Characterization of a fluoroscopic imaging system for kV and MV radiography. AB - An on-line kilovoltage (kV) imaging system has been implemented on a medical linear accelerator to verify radiotherapy field placement. A kV x-ray tube is mounted on the accelerator at 90 degrees to the megavoltage (MV) source and shares the same isocenter. Nearly identical CCD-based fluoroscopic imagers are mounted opposite the two x-ray sources. These systems are being used in a clinical study of patient setup error that examines the advantage of kV imaging for on-line localization. In the investigation reported here, the imaging performance of the kV and MV systems are characterized to provide support to the conclusions of the studies of setup error. A spatial-frequency-dependent linear systems model is used to predict the detective quantum efficiencies (DQEs) of the two systems. Each is divided into a series of gain and spreading stages. The parameters of each stage are either measured or obtained from the literature. The model predicts the system gain to within 7% of the measured gain for the MV system and to within 10% for the kV system. The systems' noise power spectra (NPSs) and modulation transfer functions (MTFs) are measured to construct the measured DQEs. X-ray fluences are calculated using modeled polyenergetic spectra. Measured DQEs agree well with those predicted by the model. The model reveals that the MV system is well optimized, and is x-ray quantum noise limited at low spatial frequencies. The kV system is suboptimal, but for purposes of patient positioning yields images superior to those produced by the MV system. This is attributed to the kV system's higher DQE and to the inherently higher contrasts present at kV energies. PMID- 10841393 TI - Lens distortion in optically coupled digital x-ray imaging. AB - The objectives of this research are to analyze geometrical distortions introduced by relay lenses in optically coupled digital x-ray imaging systems and to introduce an algorithm to correct such distortions. METHODS: The radial and tangential errors introduced by a relay lens in digital x-ray imaging were experimentally measured, using a lens-coupled CCD (charge coupled device) prototype. An algorithm was introduced to correct these distortions. Based on an x-ray image of a standard calibration grid, the algorithm first identified the location of the optical axis, then corrected the radial and tangential distortions using polynomial transformation technique. RESULTS: Lens distortions were classified and both radial and tangential distortions introduced by lenses were corrected using polynomial transformation. For the specific lens-CCD prototype investigated, the mean positional error caused by the relay lens was reduced by the correction algorithm from about eight pixels (0.69 mm) to less than 1.8 pixels (0.15 mm). Our investigation also shows that the fourth order of polynomial for the correction algorithm provided the best correction result. CONCLUSIONS: Lens distortions should be considered in position-dependent, quantitative x-ray imaging and such distortions can be minimized in CCD x-ray imaging by appropriate algorithm, as demonstrated in this paper. PMID- 10841394 TI - Photon scatter in portal images: accuracy of a fluence based pencil beam superposition algorithm. AB - The accuracy of a pencil beam algorithm to predict scattered photon fluence into portal imaging systems was studied. A data base of pencil beam kernels describing scattered photon fluence behind homogeneous water slabs (1-50 cm thick) at various air gap distances (0-100 cm) was generated using the EGS Monte Carlo code. Scatter kernels were partitioned according to particle history: singly scattered, multiply-scattered, and bremsstrahlung and positron annihilation photons. Mean energy and mean angle with respect to the incident photon pencil beam were also scored. This data allows fluence, mean energy, and mean angular data for each history type to be predicted using the pencil beam algorithm. Pencil beam algorithm predictions for 6 and 24 MV incident photon beams were compared against full Monte Carlo simulations for several inhomogeneous phantoms, including approximations to a lateral neck, and a mediastinum treatment. The accuracy of predicted scattered photon fluence, mean energy, and mean angle was investigated as a function of air gap, field size, photon history, incident beam resolution, and phantom geometry. Maximum errors in mean energies were 0.65 and 0.25 MeV for the higher and lower energy spectra, respectively, and 15 degrees for mean angles. The ability of the pencil beam algorithm to predict scatter fluence decreases with decreasing air gap, with the largest error for each phantom occurring at the exit surface. The maximum predictive error was found to be 6.9% with respect to the total fluence on the central axis. By maintaining even a small air gap (approximately 10 cm), the error in predicted scatter fluence may be kept under 3% for the phantoms and beam energies studied here. It is concluded that this pencil beam algorithm is sufficiently accurate (using International Commission on Radiation Units and Measurements Report No. 24 guidelines for absorbed dose) over the majority of clinically relevant air gaps, for further investigation in a portal dose prediction algorithm. PMID- 10841395 TI - Slit x-ray beam primary dose profiles determined by analytical transport of Compton recoil electrons. AB - Accurate measurement of radiation beam penumbras is essential for conformal radiotherapy. For this purpose a detailed knowledge of the dosimeter's spatial response is required. However, experimental determination of detector spatial response is cumbersome and restricted to the specific detector type and beam spectrum used. A model has therefore been developed to calculate in slit beam geometry both dose profiles and detector response profiles. Summations over representative photon beam spectra yield profiles for polyenergetic beams. In the present study the model is described and resulting dose profiles verified. The model combines Compton scattering of incident photons, transport of resulting electrons by Fermi-Eyges small-angle multiple scattering theory, and functions to limit electron transport. This analytic model thus yields line spread kernels of primary dose in a water phantom. It is shown that the spatial response of an ideal point detector to a primary photon beam can be well described by the model; the calculations are verified by measurements with a diamond detector in a telescopic slit geometry in which all dose contributions except for the primary dose can be excluded. Effects of photon detector behavior, source size of the linear accelerator (linac) and detector size are studied. Measurements show that slit dose profiles calculated by means of the kernel are accurate within 0.1 mm of the full-width at half-maximum. For a theoretical point source and point detector combined with a 0.2 mm wide slit, the full-width half-maximum values of the slit beam dose profiles are calculated as 0.37 mm and 0.42 mm in a 6 MV and 25 MV x-ray beam, respectively. The present study shows that the model is adequate to calculate local dose effects that are dominated by approximately mono directional, primary photon fluence. The analytic model further provides directional electron fluence information and is designed to be applied to various detectors and linac beam spectra. PMID- 10841396 TI - Patient-dependent beam-modifier physics in Monte Carlo photon dose calculations. AB - Model pencil-beam on slab calculations are used as well as a series of detailed calculations of photon and electron output from commercial accelerators to quantify level(s) of physics required for the Monte Carlo transport of photons and electrons in treatment-dependent beam modifiers, such as jaws, wedges, blocks, and multileaf collimators, in photon teletherapy dose calculations. The physics approximations investigated comprise (1) not tracking particles below a given kinetic energy, (2) continuing to track particles, but performing simplified collision physics, particularly in handling secondary particle production, and (3) not tracking particles in specific spatial regions. Figures of-merit needed to estimate the effects of these approximations are developed, and these estimates are compared with full-physics Monte Carlo calculations of the contribution of the collimating jaws to the on-axis depth-dose curve in a water phantom. These figures of merit are next used to evaluate various approximations used in coupled photon/electron physics in beam modifiers. Approximations for tracking electrons in air are then evaluated. It is found that knowledge of the materials used for beam modifiers, of the energies of the photon beams used, as well as of the length scales typically found in photon teletherapy plans, allows a number of simplifying approximations to be made in the Monte Carlo transport of secondary particles from the accelerator head and beam modifiers to the isocenter plane. PMID- 10841397 TI - Intensity modulation delivery techniques: "step & shoot" MLC auto-sequence versus the use of a modulator. AB - Two intensity modulation radiotherapy (IMRT) delivery systems, the "step & shoot" multileaf collimator (MLC) auto-sequence and the use of an intensity modulator, are compared with emphasis on the dose optimization quality and the treatment irradiation time. The intensity modulation (IM) was created by a dose gradient optimization algorithm which maximizes the target dose uniformity while maintaining dose to critical structures below a set tolerance defined by the user in terms of either a single dose value or a dose volume histogram curve for each critical structure. Two clinical cases were studied with and without dose optimization: a three-field sinus treatment and a six-field nasopharyngeal treatment. The optimization goal of the latter case included the sparing of several nearby normal structures in addition to the target dose uniformity. In both cases, the target dose uniformity initially improved quickly as the IM level increased to 5, then started to approach saturation when the MLC technique was used. In the absence of the both space and intensity discreteness intrinsic to the MLC technique, the modulator technique produced greater tumor dose uniformity and normal structure sparing. The latter showed no systematic improvement with increasing IM level using the MLC technique. For the sinus tumor treatment of 2 Gy the treatment irradiation time of the modulator technique is no more than that of the conventional treatment. For the MLC technique the irradiation time increased rapidly from 4.4 min to 12.4 min as the IM level increased from 2 to 10. Both clinical cases suggested that an IM level of 5 offered a good compromise between the dose optimization quality and treatment irradiation time. We showed that a realistic photon source model is necessary for dose computation accuracy in the MLC-IM treatments. PMID- 10841398 TI - Dose calculation and verification of intensity modulation generated by dynamic multileaf collimators. AB - While the development of inverse planning tools for optimizing dose distributions has come to a level of maturity, intensity modulation has not yet been widely implemented in clinical use because of problems related to its practical delivery and a lack of verification tools and quality assurance (QA) procedures. One of the prerequisites is a dose calculation algorithm that achieves good accuracy. The purpose of this work was twofold. A primary-scatter separation dose model has been extended to account for intensity modulation generated by a dynamic multileaf collimator (MLC). Then the calculation procedures have been tested by comparison with carefully carried out experiments. Intensity modulation is being accounted for by means of a 2D (two-dimensional) matrix of correction factors that modifies the spatial fluence distribution, incident to the patient. The dose calculation for the corresponding open field is then affected by those correction factors. They are used in order to weight separately the primary and the scatter component of the dose at a given point. In order to verify that the calculated dose distributions are in good agreement with measurements on our machine, we have designed a set of test intensity distributions and performed measurements with 6 and 20 MV photons on a Varian Clinac 2300C/D linear accelerator equipped with a 40 leaf pair dynamic MLC. Comparison between calculated and measured dose distributions for a number of representative cases shows, in general, good agreement (within 3% of the normalization in low dose gradient regions and within 3 mm distance-to-dose in high dose gradient regions). For absolute dose calculations (monitor unit calculations), comparison between calculation and measurement reveals good agreement (within 2%) for all tested cases (with the condition that the prescription point is not located on a high dose gradient region). PMID- 10841399 TI - A dosimetric leaf-setting strategy for shaping radiation fields using a multileaf collimator. AB - A dosimetric leaf-setting strategy of using multileaf collimators (MLC) for shaping radiation fields has been developed. Existing MLC leaf-setting strategies are all based upon geometric criteria. This new approach, however, matches a prescribed field contour with a MLC using clinically consistent dosimetric criteria. The leaf positions are determined using an iterative optimization algorithm. An empirical dose model was developed to compare the dosimetric-based leaf-setting strategy with the geometric-based leaf-setting strategies. Differences up to half a centimeter in the leaf positions and isodose lines were found between setting the MLC geometrically and setting the MLC dosimetrically. The dosimetric leaf-setting strategy provides the ability to achieve better dose conformation for a clinically desired isodose line. Since the desired isodose line that covers a treatment volume is typically higher than 50% of the maximum dose, the scalloping effects due to the finite leaf width at the leaf edge or 50% isodose lines are much reduced. Another benefit of the dosimetric leaf-setting is that it separates the leaf-setting process from the treatment planning process, and this frees the treatment planning vendors from developing detailed dose models for various existing types and future upgrades of MLC systems. PMID- 10841400 TI - A practical technique for verification of three-dimensional conformal dose distributions in stereotactic radiosurgery. AB - The trend toward conformal techniques in stereotactic radiosurgery necessitates an accurate and practical method for verification of irregular three-dimensional dose distributions. This work presents the design and evaluation of a phantom system facilitating the measurement of conformal dose distributions using one or more arrays of up to 20 radiographic films separated by 3.2 mm-thick tissue equivalent spacers. Using Electron Gamma Shower version 4 (EGS4) Monte Carlo simulation, we show that for 6 MV radiosurgical photon beams this arrangement preserves tissue-equivalence to within 1%. The phantom provides 0.25 mm in-plane spatial resolution and multiple sets of films may be used to resample the dose volume in orthogonal planes. Dedicated software has been developed to automate the process of ordering and orienting of scanned film images, conversion of scanned pixel value to dose, resampling of one or more sets of film images and subsequent export of images in DICOM format for coregistration of planned and measured dose volumes. Calculated and measured isodose surfaces for a simple, circular-beam treatment agree to within 1.5 mm throughout the dose volume. For conformal radiosurgical applications, the measured and planned dose distributions agree to within the uncertainty of the manufacture of irregularly shaped collimators. The sensitivity of this technique to minor spatial inaccuracies in beam shaping is also demonstrated. PMID- 10841401 TI - Truncation of blood curves to enhance imaging and therapy with monoclonal antibodies. AB - Targeting of monoclonal antibody (Mab) to solid tumor sites is a function of the blood curve of activity versus time. It has been suggested that the blood curve be artificially reduced to approach zero so that the contrast between tumor and blood uptake is maximized. We analyzed tumor uptake as a function of the time tc of blood curve truncation. By using a convolution approach, we were able to find the optimal times for setting the blood curve to zero in either diagnostic or therapeutic animal examples. Two iodinated cT84.66 anti-CEA engineered fragments, diabody and minibody, were considered using previous data from nude mouse studies involving the LS174T colorectal tumor model. Figures of merit (FOMs) were used to compare ordinary and truncated blood curves and their associated tumor accumulations. Using a 1231 label, it was seen that the appropriate time for diagnostic truncation occurred when tumor uptake, as measured, was a maximum. The corresponding point for therapy (with 1311 as a label) was at infinite time. We also demonstrated that the use of traditional indices led to ambiguities in the choice of truncation times. The traditional therapy index, the ratio of the integral of the tumor uptake to the integral of the blood uptake, was found to be a numerical constant independent of tc. This ratio was proved to be the integral of the tumor impulse response function. Use of such convolution techniques to assess truncation of the perfused material is probably also applicable to multistep processes as well as to lesion targeting with other tumor-specific pharmaceuticals. PMID- 10841402 TI - Optimization of radiosurgery treatment planning via mixed integer programming. AB - An automated optimization algorithm based on mixed integer programming techniques is presented for generating high-quality treatment plans for LINAC radiosurgery treatment. The physical planning in radiosurgery treatment involves selecting among a large collection of beams with different physical parameters an optimal beam configuration (geometries and intensities) to deliver the clinically prescribed radiation dose to the tumor volume while sparing the nearby critical structure and normal tissue. The proposed mixed integer programming models incorporate strict dose restrictions on tumor volume, and constraints on the desired number of beams, isocenters, couch angles, and gantry angles. The model seeks to deliver full prescription dose coverage and uniform radiation dose to the tumor volume while minimizing the excess radiation to the periphery normal tissue. In particular, it ensures that proximal normal tissues receive minimal dose via rapid dose fall-off. Preliminary numerical tests on a single patient case indicate that this approach can produce exceptionally high-quality plans in a fraction of the time required using the procedure currently employed by clinicians. The resulting plans provide highly uniform prescription dose to the tumor volume while drastically reducing the irradiation received by the proximal critical normal tissue. PMID- 10841403 TI - Application of radiosurgery principles to a target in the breast: a dosimetric study. AB - PURPOSE: To investigate the technical and physical feasibility of using a radiosurgery-like technique to irradiate a small target within the breast with a single fraction. MATERIAL AND METHODS: During diagnostic biopsy, a tantalum surgical clip is placed in the lesion identified at mammography. Transverse CT scans over the entire breast are obtained, as the patient lies prone on a special table that allows the breast to hang down. The clip is used as a reference point to define the isocenter of the radiation treatment. RESULTS: The clip is visible on port films taken with a 4 MV beam, allowing the isocenter to be set to its planned location. No movement of the hanging breast is visually detected. The possible beam directions are enclosed by a 220 degrees horizontal x 180 degrees vertical angular interval. Dosimetry of two "radiosurgical" examples, (A) seven fixed horizontal beams and (B) six 45 degrees arcs and a 90 degrees sagittal arc using a 4 MV x-ray beam with a 32 mm diameter collimator, are discussed. Both field arrangements produce adequate tumor coverage: the minimum target dose is 83% of the dose maximum in the fixed beam arrangement and 86% in the multiarc setup. In arrangement A the lung and other tissues external to the breast receive dose only from scattered radiation. In arrangement B the maximum lung dose is less than 5% of the dose to isocenter. CONCLUSION: From a dosimetric point of view both described techniques are feasible, and the radiosurgery-like treatment is executable. PMID- 10841404 TI - A systematic evaluation of air cavity dose perturbation in megavoltage x-ray beams. AB - The EGS4 Monte Carlo radiation transport code was used to systematically study the dose perturbation near planar and cylindrical air cavities in a water medium irradiated by megavoltage x-ray beams. The variables of the problem included x ray energy, cavity shape and dimension, and depth of the cavity in water. The Monte Carlo code was initially validated against published measurements and its results were found to agree within 2% with the published measurements. The study results indicate that the dose perturbation is strongly dependent on x-ray energy, field size, depth, and size of cavity in water. For example, the Monte Carlo calculations show dose reductions of 42% and 18% at 0.05 and 2 mm, respectively, beyond the air-water interface distal to the radiation source for a 3 cm thick air slab irradiated by a single 5x5 cm2 15 MV beam. The dose reductions are smaller for a parallel-opposed pair of 5x5 cm2 15 MV x-ray beams, being 21% and 11% for the same depths. The combined set of Monte Carlo calculations showed that the dose reduction near an air cavity is greater for: (a) Smaller x-ray field size, (b) higher x-ray energy, (c) larger air-cavity size, and (d) smaller depth in water where the air cavity is situated. A potential clinical application of these results to the treatment of prostate cancer is discussed. PMID- 10841405 TI - A method to check the accuracy of dose computation using quality index: application to scatter contribution in high energy photon beams. AB - Computerized dose calculation verification is a relevant component of radiotherapy treatment planning quality assurance. The usual procedure is to compare measurements to computations for several standard situations. As cases become more complex, special test phantoms and beam arrangements must be used, and an experimental procedure must be carefully established. In this paper we follow a new methodology to prepare a set of reference data that may be used to verify the accuracy of dose calculations involving changes in the scatter component of photon beams. The advantage of this methodology is that local measurements are not required. A quantitative evaluation of dose modifications was performed by means of correction factors (CF). For this purpose, three geometrical configurations were designed (asymmetric, symmetric, and reference) where the primary component was kept constant and the scatter component was varied by changing the height (h) of lateral columns. Measurements were performed in polystyrene phantoms for seven photon beam energies. CF were derived as the ratio of the absolute dose measured at the point of interest to the absolute dose for the reference configuration, for the asymmetric and symmetric configurations, respectively. They were expressed as a function of beam quality (QI). We have verified that, for all configurations studied, CF decrease with QI. For h = 15 cm, CF remain practically constant, whatever machine technology is used [the mean values of CF for the asymmetric and symmetric cases are CFa= 1.028 (0.2% 1 s.d.) and CFs= 1.058 (0.4% 1 s.d.)]. We have developed a test protocol and we have chosen those configurations corresponding to h = 15 cm because they both present greater values of the CF and lower standard deviations. The direct application of the method is straightforward. The user can reproduce on his local TPS the three experimental configurations described in the test protocol, and then compute CF which can be compared to our reference data set for any beam quality. PMID- 10841406 TI - In-phantom response of LiF TLD-100 for dosimetry of 192Ir HDR source. AB - An experiment was carried out to reevaluate the response of LiF TLD-100 rods (1 mmx1 mmx6 mm) at different depths in a water substituting phantom to provide an answer to a prevailing controversy about the over-response of LiF to the softened photon spectra of 192Ir HDR source at depths in phantom due to its photon energy dependence. Claims of some authors that LiF TLDs over-responds by 8.5% at 10 cm depth in phantom, necessitating depth-dependent correction factors even for an 192Ir source and of some others for no over-response were evaluated. The over response of LiF TLD-100 rods, against a calibrated ion chamber having a photon energy-independent response within 2%, was found to be not exceeding 2.5% at a depth of 10 cm in the phantom as compared to a depth at 1 cm, for a precision of the order of +/- 1% (1sigma) in the TLD measurements. By using ISO equivalent photon beams, photon energy dependence of the dosimeters was evaluated and for LiF TLD-100 rods it was found to be in close agreement (within 3%) with the ratios of mass energy absorption coefficients of LiF and water in the range of effective photon energy from 26 keV to 1.25 MeV. Parameters that could contribute to the discrepancy in the reported values of experimental results have been discussed. PMID- 10841407 TI - A manual algorithm for computing dwell times for two-catheter endobronchial treatments using HDR brachytherapy. AB - A method has been developed to permit the calculation of dwell times for endobronchial high dose rate (HDR) brachytherapy treatment using two catheters, without using a dedicated treatment planning system. Worksheets were developed to guide a user through manual calculations, and a computer program was written to automate the process. This empirical algorithm produces dose distributions that are clinically safe and reasonable. The total dwell times match those predicted by individually optimized distributions to within a few percent. The method has been used most frequently as a quality assurance check on optimized plans produced by a commercial treatment planning system, but it can serve as a back-up method should the commercial system fail. [Worksheets with example calculations may be retrieved using anonymous ftp from the American Institute of Physics, Electronic Physics Auxillary Publication Service (EPAPS).] PMID- 10841408 TI - Generation of uniformly distributed dose points for anatomy-based three dimensional dose optimization methods in brachytherapy. AB - We have studied the accuracy of statistical parameters of dose distributions in brachytherapy using actual clinical implants. These include the mean, minimum and maximum dose values and the variance of the dose distribution inside the PTV (planning target volume), and on the surface of the PTV. These properties have been studied as a function of the number of uniformly distributed sampling points. These parameters, or the variants of these parameters, are used directly or indirectly in optimization procedures or for a description of the dose distribution. The accurate determination of these parameters depends on the sampling point distribution from which they have been obtained. Some optimization methods ignore catheters and critical structures surrounded by the PTV or alternatively consider as surface dose points only those on the contour lines of the PTV. D(min) and D(max) are extreme dose values which are either on the PTV surface or within the PTV. They must be avoided for specification and optimization purposes in brachytherapy. Using D(mean) and the variance of D which we have shown to be stable parameters, achieves a more reliable description of the dose distribution on the PTV surface and within the PTV volume than does D(min) and D(max). Generation of dose points on the real surface of the PTV is obligatory and the consideration of catheter volumes results in a realistic description of anatomical dose distributions. PMID- 10841409 TI - Catheter autoreconstruction in computed tomography based brachytherapy treatment planning. AB - The aim of this study is to develop an automatic reconstruction of brachytherapy catheters using CT (computed tomography) data. Previously no such automatic facility has existed in any treatment planning software. To achieve this facility we have developed tools for the automatic reconstruction (which we term autoreconstruction) of plastic and metallic catheters. These algorithms overcome a number of difficulties which arise when a large number of catheters are present. These include situations with intersecting catheters and with loop techniques. The time required for the catheter reconstruction process using our autoreconstruction method is significantly reduced. The accuracy of our autoreconstruction is at least as high as the classical manual slice-by-slice method. PMID- 10841410 TI - Dosimetric effects of needle divergence in prostate seed implant using 125I and 103Pd radioactive seeds. AB - In prostate seed implants, radioactive seeds are implanted into the prostate through a guiding needle with the help of a template and real-time imaging. The ideal locations of the guiding needles and the relative positions of the seeds in each needle are determined before the implantation under the assumption that the needles inserted at different locations will remain parallel. In actual implantation, the direction of the needle is subject variation. In this work, we studied how the dosimetry quality of an implant may be affected when the guiding needles deviate from its planned orientations. Needle divergence of varying degree was simulated on spherical models and actual patient implants. It was found that needle divergence degraded the dosimetric quality of an implant: The minimum target dose, the target dose coverage and therefore the tumor biological effective dose were quantitatively decreased as compared to the reference implant. The magnitude of degradation increased almost linearly with respect to the magnitude of needle divergence. For iodine-125 implants, the average reduction in minimum target dose was about 10% and 20% for needle divergence of standard deviation of 5(0) and 10(0), respectively. The dose coverage in the target was reduced by about 1% and 3% for needle divergence of standard deviation of 5(0) and 10(0), respectively. Implants designed with palladium-103 showed additional 5% reduction in minimum target dose while the effect on dose coverage was about the same as compared to the iodine-125 implants. The degree of dosimetry degradation was shown to be dependent on the size of target volume, the seed spacing used, the use of seeding margin, and on the actual configuration of needle orientations in a given implant. One needs to minimize the physical causes of needle divergence in order to minimize its impact on planned dosimetry. The study suggests that the displacement between a needle image and its planned grid point at the base of prostate should be kept less than 5 mm in order to minimize the reduction in D(min)(<5%) and the increase in cell-survival (< a factor of 10) from the planned dosimetry. PMID- 10841411 TI - Isotope choice and the effect of edema on prostate brachytherapy dosimetry. AB - In prostate brachytherapy, post implant dosimetry quality parameters may be strongly affected by edema brought on by the trauma of the implant procedure since the amount of edema and the time course of its resolution are highly variable from patient to patient. Edema was simulated from preplans on three prostates which had ultrasound prostate volumes of 18.7, 40.7 and 60.2 cm3 expanded to planning volumes of 32.9, 60.0 and 87.8 cm3, respectively. The preplans were designed so that identical seed distributions for a given prostate gave virtually identical target dose coverage of 99.7+/-0.3% of the planning volume when using either 125I or 103Pd. Simulated CT edema volume expansions of 0%, 10%, 20% and 30% were imposed anisotropically in accordance with clinical observations so that the expansion in the superior-inferior direction was twice that of the transverse dimensions. Dose-volume histograms (DVHs) were analyzed for each prostate as a function of isotope and degree of edema. The 103Pd implants were more greatly affected by fixed amounts of edema than 125I implants, and the slopes of the DVH curves indicate less homogeneity from 103Pd implants. The DVHs were then weighted according to the portion of the isotope decay curve occupied by each edema step for half-lives of edema resolution of 5, 10 and 20 days which are within the range of clinically observed resolution times. The weighted DVHs were summed to give a net DVH corresponding to the overall dynamic effect of edema. A greater fraction of the defined prostate volume received doses in the range of likely therapeutic significance, from 75% to 125% of the prescribed minimal peripheral dose (mPD), from 125I implants than from 103Pd implants. These differences in dosimetric quality arise from two differences in the physical properties of the isotopes: more rapid attenuation of 103Pd photons with distance creates cool spots in an edematous prostate, and the shorter half life of 103Pd causes a greater fraction of the isotope decay to consist of the prostate in an edematous state. An increase in 103Pd seed strength by about 10% beyond that required to achieve equal coverage with an identical seed distribution using 125I should minimize the differences brought on by edema. PMID- 10841412 TI - Monte Carlo-aided dosimetry of the Symmetra model I25.S06 125I, interstitial brachytherapy seed. AB - A dosimetric study of a new 125I seed for permanent prostate implant, the Symmetra 1251 Seed model I25.S06, has been undertaken utilizing Monte Carlo photon transport calculations. All dosimetric quantities recommended by the AAPM Task Group 43 (TG-43) report have been calculated. Quantities determined are dose rate constant, radial dose function, anisotropy function, anisotropy factor, and anisotropy constant. The recently (January 1999) revised NIST (National Institute of Standards and Technology) 1251 standard for air kerma strength calibration was taken into account as well as updated interaction cross-section data. Calculations were done for the competing model 6702 source for the purpose of comparison. The calculated dose-rate constants for the two seeds are 1.010 and 1.016 cGyh(-1)U(-1) for the Symmetra and model 6702 seeds, respectively. The latter value deviates from the value, 1.039 cGyh(-1)U(-1), recommended in the TG 43 report. The calculated radial dose function for the Symmetra new seed is more penetrating than that of the model 6711 seed (by 20% at 5 cm distance) but agrees closely (within statistical errors) with that of the model 6702 seed up to distances of 10 cm. The anisotropy function for the seed is also close to that for the 6702 seed with a tendency of somewhat more pronounced anisotropy (lower values at small angles from the longitudinal axis). Compared to the model 6711 seed, the Symmetra new seed is more isotropic. The anisotropy constants (the anisotropy function averaged with respect to angle and distance) for the three seed models are within 2%. PMID- 10841413 TI - Monte Carlo modeling of radiation dose distributions in intravascular radiation therapy. AB - Radiation dose distributions are developed for balloon and wire sources of radioactivity within coronary arteries. The Monte Carlo codes MCNP 4B and EGS4 were used to calculate dose distributions for photons and electrons at discrete energies around such sources, with and without the presence of a high-density atherosclerotic plaque. An interactive computer program was developed which then calculates dose distributions for many radionuclides by applying the emission spectra to the discrete energy grids calculated by the Monte Carlo codes, weighting appropriately for electron energy and abundance. Results for Re-186 and Re-188 balloon sources are shown in comparison to an Ir-192 wire source. The program provides dose distributions as well as estimates of activity levels needed to deliver prescribed doses to the vessel wall at selected distances from the lumen in a selected time interval. In addition, dose calculations are presented in this paper for other organs in the body, from photon radiation as well as from possible loss of liquid activity into the bloodstream in the case of a balloon rupture. These results, especially the interactive computer program permitting easy comparison of various radionuclides and their physical characteristics, will greatly facilitate the comparison process and aid in the selection of the best candidate(s) for clinical use. PMID- 10841414 TI - Dosimetric characteristics of the InterSource103 palladium brachytherapy source. AB - 103Pd brachytherapy sources are being used for interstitial implants in tumor sites such as the prostate. Recently, the InterSource103 palladium source has been introduced, which has a design different from that of other sources presently on the market. Dosimetric characteristics (i.e., dose rate constant, radial dose function, and anisotropy function) of this source were experimentally and theoretically determined following the AAPM Task Group 43 (TG-43) recommendations and were related to the 1999 NIST calibration assigned to this source [Sk, 99std]. Measurements were performed in a solid water phantom using LiF thermoluminescent dosimeters. The measured data was compared with Monte Carlo simulations performed in solid water using the PTRAN code. The calculations were then performed in liquid water to obtain the dosimetric information for clinical applications as per TG-43 recommendation. The results indicated that the dose rate constant, lambda, of the InterSource103 palladium source was 0.664+/-5% cGy/h/U using TLD measurements and 0.660+/-3% cGy/h/U using Monte Carlo simulations in solid water. The calculated value in liquid water was found to be 0.696 +/- 3 % cGy/h/U. The radial dose function, g(r), of the new 103Pd source was measured at distances ranging from 0.5 to 10 cm using LiF TLD in solid water phantom material. The Monte Carlo simulations were performed at distances ranging from 0.1 to 10 cm from the source center in solid water and liquid water. The anisotropy function, F(r, theta), was measured at distances of 2, 3, 5, and 7 cm from the source center and calculated at distances of 0.5, 1, 2, 3, 5, and 7 cm from the source center. Complete dosimetric data are described in this paper. Per TG-43, the values reported in water should be used for clinical treatment planning systems. PMID- 10841415 TI - High resolution computed tomography and MRI for monitoring lung tumor growth in mice undergoing radioimmunotherapy: correlation with histology. AB - A model lung tumor system has been developed in mice for the evaluation of vascular targeted radioimmunotherapy. In this model, EMT-6 mammary carcinoma tumors growing in the lung are treated with 213Bi, an alpha particle emitter, which is targeted to lung blood vessels using a monoclonal antibody. Smaller tumors (< 100 microm in diameter) are cured, but larger tumors undergo a period of regression and then regrow and ultimately prove lethal. The goal of this work was to determine if external imaging with MRI or CT could be used routinely to monitor the growth/ regression of lung tumors in live mice. To attempt to evaluate individual tumors in vivo, animals were initially imaged with magnetic resonance imaging (MRI). High resolution MRI images could be obtained only after sacrifice when lungs were not moving. In contrast, high resolution computed tomography (CT) produced evaluable images from anesthetized animals. Serial CT images (up to 5/animal) were collected over a 17 day period of tumor growth and treatment. When tumored animals became moribund, animals were sacrificed and lungs were inflated with fixative, embedded in paraffin, and then sectioned serially to compare the detection of tumors by high resolution CT with detection by histology. CT proved most useful in detecting lung tumors located in the hilar area and least useful in detecting serosal surface and anterior lobe tumor foci. Overall, CT images of live animals revealed tumors in approximately 2/3 of cases detected in histologic serial sections when relatively few tumors were present per lung. Detection of lesions and their resolution post therapy were complicated due to residual hemorrhagic, regressing tumor nodules and the development of lung edema both of which appeared as high density areas in the CT scans. We conclude that the microCT method used could identify some lung tumors as small as 100 microm in diameter; however, no concrete evaluation of therapy induced regression of the tumors could be made with CT analyses alone. PMID- 10841416 TI - Monte Carlo calculations and experimental measurements of dosimetry parameters of a new 103Pd source. AB - Permanent prostate implantation using 125I (iodine) or 103Pd (palladium) sources is a popular treatment option in the management of early prostate cancer. As sources of new designs are developed and marketed for application in permanent prostate implantations, their dosimetric characteristics must be carefully determined in order to maintain the accuracy of patient treatment. This report presents the results of experimental measurements and Monte Carlo calculations of the dosimetric parameters performed for a newly available 103Pd seed source. The measurements were performed in a large scanning water phantom using a diode detector. The positioning of the source and detector was achieved by a computer controlled positioning mechanism in the scanning water phantom. The dose rate constant in water for the new 103Pd source was determined from measurements with the diode detector calibrated with 125I sources of known air-kerma strength. The radial dose function values for the source were measured using the diode detector. Monte Carlo photon transport calculations were then used to calculate the dosimetric parameters of dose rate constant, radial dose function, and anisotropy function using an accurate geometric model of the source. The measured dose rate constant of 0.693 cGy/U-hr compares well with the Monte Carlo calculated value of 0.677 cGy/U-hr. These results are further compared with data on existing 103Pd sources. PMID- 10841417 TI - What is the best proton energy for accelerator-based BNCT using the 7Li(p,n)7Be reaction? AB - With a growing interest in the use of accelerator-based epithermal neutron sources for BNCT programs, in particular those based upon the 7Li(p,n)7Be reaction, there is a need to address the question of "what is the best proton energy to use?" This paper considers this question by using radiation transport calculations to investigate a range of proton energies from 2.15 to 3.5 MeV and a range of moderator sizes. This study has moved away completely from the use of empty therapy beam parameters and instead defines the beam quality and optimizes the moderator design using widely accepted in-phantom treatment planning figures of merit. It is concluded that up to a proton energy of about 2.8 MeV there is no observed variation in the achievable therapy beam quality, but a price is paid in terms of treatment time for not choosing the upper limit of this range. For higher proton energies, the beam quality falls, but with no improvement in treatment time for optimum configurations. PMID- 10841418 TI - Radiobiological significance of beamline dependent proton energy distributions in a spread-out Bragg peak. AB - Similar target doses can be achieved with different mixed radiation fields, i.e., particle energy distributions, produced by a practical proton beam and a range modulator. The dose delivered in particle therapy can be described as the integral of fluence times the total mass stopping power over the particle energy distributions. We employed Monte Carlo simulations to explore the influence on the relative biological effectiveness (RBE) of the energy and the energy spread of the proton beam incident on a range modulator system. Using different beams, the conditions of beam delivery were adjusted so that similar spread out Bragg peak (SOBP) doses were delivered to a simulated water phantom. We calculated the RBE for inactivation of three different cell lines using the track structure model. The RBE depends on the details of the dose deposition and the biological characteristics of the irradiated tissue. Our calculations show that, for differing beam conditions, the corresponding differences in the total mass stopping power distributions are reflected in differences in the RBE. However, these differences are remarkable only at the very distal edge of the SOBP, for low doses, and/or for large differences in beam setup. PMID- 10841419 TI - A variable speed translating couch technique for total body irradiation. AB - We have developed a variable speed translating patient couch system for the delivery of total body irradiation (TBI). For a whole body Rando-type phantom, dose variation at mid-plane relative to the prescription point (navel) can be as high as 15% (neck or legs) with a constant velocity. By taking into account variations in body thickness, the intensity modulation radiation therapy, resulting from variable velocities, effectively delivers a uniform dose distribution at mid plane. The couch control user interface, technical aspects and dose planning optimization procedure for determining velocity distribution are described. PMID- 10841420 TI - Theoretical study of temperature elevation at muscle/bone interface during ultrasound hyperthermia. AB - This paper examines the distributions of the SAR (specific absorption rate) ratio and temperature elevation when an ultrasound beam propagates through the interface of muscle and bone. This interface is regarded as a flat boundary to partition the energy of the ultrasound beam, and the analytical solution of temperature distribution is based on the steady-state bio-heat transfer equation. The parameters considered are the incident angle of ultrasound beam, the ultrasound frequency, the acoustic attenuation coefficients of refracted longitudinal and shear waves in bone, and the blood perfusion in muscle. The results show that the peak of the SAR ratio is always at the interface of muscle and bone, while the peak of temperature is located in the bone region beyond the interface. A muscle with lower perfusion or a bone with higher acoustic attenuation results in the shifting of the temperature peak closer to the interface. It is more difficult to heat a higher perfused muscle in front of a bone using a lower frequency ultrasound since the temperature elevation for bone relative to muscle is greater. PMID- 10841421 TI - Magnetic resonance imaging of thermal coagulation effects in a phantom for calibrating thermal therapy devices. AB - A material has been developed and tested that permanently records thermal response patterns from heating devices. The material consists of a mixture of polyacrylamide and 18% w/w bovine serum albumin. Thermal denaturation is complete when the local temperature exceeds 70 degrees C, causing a large reduction in the T2 of the material. Three-dimensional distributions of "thermal damage" can be assessed using standard magnetic resonance imaging sequences. The material works well with microwave heating devices and is adaptable for use with ultrasound, radio-frequency, or laser heating devices. Suggested uses include characterizing heating devices prior to treatment and developing new clinical applications for thermal therapies. PMID- 10841422 TI - Saturation current and collection efficiency for ionization chambers in pulsed beams. AB - Saturation currents and collection efficiencies in ionization chambers exposed to pulsed megavoltage photon and electron beams are determined assuming a linear relationship between 1/I and 1/V in the extreme near-saturation region, with I and V the chamber current and polarizing voltage, respectively. Careful measurements of chamber current against polarizing voltage in the extreme near saturation region reveal a current rising faster than that predicted by the linear relationship. This excess current combined with conventional "two-voltage" technique for determination of collection efficiency may result in an up to 0.7% overestimate of the saturation current for standard radiation field sizes of 10X10 cm2. The measured excess current is attributed to charge multiplication in the chamber air volume and to radiation-induced conductivity in the stem of the chamber (stem effect). These effects may be accounted for by an exponential term used in conjunction with Boag's equation for collection efficiency in pulsed beams. The semiempirical model follows the experimental data well and accounts for both the charge recombination as well as for the charge multiplication effects and the chamber stem effect. PMID- 10841423 TI - New water equivalent liquid scintillation solutions for 3D dosimetry. AB - Despite recent advances in radiochromic film and gel dosimetry techniques, radiation therapy still lacks an efficient, accurate, and convenient dose measurement method capable of measuring the dose simultaneously over a plane or a volume (3D). A possibility for creating such a 3D method based on observing scintillation photons emitted from an irradiated volume was recently reported [A. S. Kirov et al., Med. Phys. 26, 1069 (1999)]. In the present article, we investigate the potential to use a liquid scintillation solution (LS) as a dose sensitive media and, simultaneously, as a water equivalent phantom material which fills the measurement volume. We show that matching water density in addition to energy absorption properties is important for using the LS solution as a phantom. Through a parametric study of the LS attenuation and absorption coefficients as well as Monte Carlo dose calculations and scintillation efficiency measurements we developed novel LS materials. For the new solutions, the calculated dose in LS is within 8% of the dose to water for depths up to 5 cm for photons having energies between 30 keV and 2 MeV. The new LS solutions, which are loaded with a Si containing compound, retain more than 85% of the scintillation efficiency of the unloaded solutions and exhibit high localization of the scintillation process. The new LS solutions are superior with respect to efficiency and water equivalence to plastic scintillator materials used in dosimetry and may be used apart from the mentioned 3D method. PMID- 10841424 TI - Comparison of trabecular bone architecture in young and old bones. AB - In this study, different parameters currently applied for the description of trabecular bone architecture in young and old subjects are compared. Moreover, new parameters are proposed and assessed. For the investigations, the two dimensional images of vertebral body sections are acquired with the use of a low magnification digital camera. The parameters were calculated both for the skeleton and for the bone/marrow interface distinguished in the images of the trabecular network. The following methods of the descriptions of the trabecular bone architecture were considered: histomorphometric analysis (BV/TV, asymmetry, mean trabeculae length), fractal geometry technique, Euler characteristic, star volume of the marrow cavity, the mean distance between two points of the trabecular network, and the probability of disconnection (is straightforwardly connected with the number of separated parts of the network). Moreover, bone mineral density (BMD) was determined for comparative purposes. The quantities directly extracted form the images are also combined to produce new indexes, which better differentiate young and old bones. It was found that the BMD, the BV/TV, the star volume, the Euler number, and the probability of disconnection might be used as indicators of the age-related changes of trabecular bone. The parameters could be measured with the precision comparable to that of the BMD. The only exception is the probability of disconnection. Highly significant correlations were demonstrated between bone density (BMD, BV/TV) and trabecular architecture (SV/GV, probability of disconnection). PMID- 10841425 TI - Depth-segmented detector for x-ray absorptiometry. AB - A new energy-dependent multi-cell detector, which is a generalization of the conventional front-back detector, was studied using computer simulations. The noise performance of the detector for bone quantitation was examined in comparison to an ideal energy discriminating detector, and front-back detectors with and without inter-detector filters. The front-back detectors were optimized for a reference object composed of water and bone, and then compared to the new detector over a range of object compositions. In this paper, precision in calculated bone thickness is used as the criterion for evaluating detector performance. Simulations show that the segmented detector always performs better than the front-back detector without an inter-detector filter. It outperforms the detector incorporating a filter by an amount that depends on the heterogeneity of the x-ray spectrum. In addition, for single component radiographic images, this multi-cell detector retains information which is lost in the front-back detector with a filter layer. PMID- 10841426 TI - Neural network based algorithm to quantify joint space width in joints of the hand for arthritis assessment. AB - Arthritis diseases are widespread with enormous societal costs. The two most common forms, rheumatoid arthritis and osteoarthritis, affect joints of the hand and cause narrowing of the joint spaces as the disease destroys the articular cartilage. Radiographic assessment is one of the most promising tools to detect subtle changes in joint space width (JSW), and therefore disease progression. Currently radiographic assessment of arthritis in joints of the hand is accomplished though semiquantitative subjective scoring systems which do not provide a quantitative measurement of the JSW. We describe here an automated method which calculates the average JSW of the metacarpophalangeal (MCP), proximal interphalangeal (PIP), and distal interphalangeal (DIP) joint spaces for fingers 2 to 5 (index, middle, ring, and little) on digitized hand radiographs. The method was tested with a set of 54 hand radiographs on joints with mild to moderate rheumatoid arthritis. Performance was evaluated by comparing algorithm measured JSW to a gold standard determined from expertly hand-drawn joint margins. The agreement was quantified by a measurement of root mean square deviation, 0.148 mm, 0.089 mm, and 0.114 mm for the MCP, PIP, and DIP joints, respectively. In addition, the algorithm measured JSW strongly correlated with the gold standard: R2=0.80 (MCP), R2= 0.82 (PIP), and R2= 0.84 (DIP). This is an accurate and robust algorithm and should provide a more quantitative measure of disease progression than current methods. PMID- 10841427 TI - Scanning thermoacoustic tomography in biological tissue. AB - Microwave-induced thermoacoustic tomography was explored to image biological tissue. Short microwave pulses irradiated tissue to generate acoustic waves by thermoelastic expansion. The microwave-induced thermoacoustic waves were detected with a focused ultrasonic transducer. Each time-domain signal from the ultrasonic transducer represented a one-dimensional image along the acoustic axis of the ultrasonic transducer similar to an ultrasonic A-scan. Scanning the system perpendicularly to the acoustic axis of the ultrasonic transducer would generate multi-dimensional images. Two-dimensional tomographic images of biological tissue were obtained with 3-GHz microwaves. The axial and lateral resolutions were characterized. The time-domain piezo-electric signal from the ultrasonic transducer in response to the thermoacoustic signal was simulated theoretically, and the theoretical result agreed with the experimental result very well. PMID- 10841428 TI - Comment on "Multileaf collimator interleaf transmission" [Med. Phys. 26, 176-186 (1999)]. PMID- 10841429 TI - Comment on "Medical physics graduate programs should focus on education and research and leave clinical training to residencies: [Med Phys 26, 2051-2053 (1999)]. PMID- 10841431 TI - Fenofibrate lowers blood pressure in two genetic models of hypertension. AB - Fenofibrate, a commonly used lipid lowering drug, induces the expression of the gene coding for cytochrome P450-4A, whose major product is 20 hydroxyeicosatetraenoic acid (20-HETE). 20-HETE, a potassium channel antagonist, could increase or decrease blood pressure (BP). We studied the effects of four weeks of oral fenofibrate on BP, urine output (UVol), plasma renin activity (PRA), and urine protein excretion in young (4-5 weeks) stroke prone spontaneously hypertensive rats (SHRSP), older (25 weeks) SHRSP, Dahl salt sensitive rats (Dahl S) on a high salt diet, Dahl S rats on a normal salt diet, and normotensive Sprague-Dawley (SD) rats. Fenofibrate prevented the increase in BP in 4-5 week old SHRSP, reduced BP in 25 week old SHRSP, but had no effect on BP in normotensive SD rats. Similarly, fenofibrate prevented the increase in BP in Dahl S rats on a high salt diet, but had no effect in Dahl S rats on a low salt diet. Fenofibrate increased UVol (and reduced weight gain) in young SHRSP and tended to increase it in other groups. It also increased PRA 2 to 5-fold in all groups except older SHRSP. Young SHRSP receiving fenofibrate excreted significantly less urine protein than control rats. The drug reduced proteinuria in Dahl S rats on high salt diet, but had no significant effect on proteinuria in other groups. In summary, fenofibrate reduced blood pressure and weight gain, increased UVol and PRA, and reduced urine protein excretion in young SHRSP. Other groups of animals showed these changes to a variable, but directionally similar extent. These findings are consistent with a natriuretic effect of fenofibrate. PMID- 10841430 TI - Effects of serotonin on the physiology of the rabbit small intestine. AB - Serotonin has been shown to alter the intestinal transport of ions and intestinal motility. These effects may interfere with each other, modulating the whole physiology of the intestine. We have previously shown that serotonin also alters the transport of nutrients. Thus, the aims of the present work were to determine the possible interference between the secretagogue effect of serotonin and the mechanism by which serotonin inhibits the absorption of nutrients, and to study the effect of serotonin on the digestive activity of nutrients of the brush border membrane jejunum enterocyte in the rabbit. The results show that the secretagogue effect of serotonin neither affects the inhibitory effect of serotonin on the intestinal absorption of the nutrients, nor affects the activity of Na+/K+-ATPase. The activity of sucrase and aminopeptidase N was also not affected by serotonin in the rabbit jejunum. Finally, we also studied different parameters of the motility in the rabbit small intestine. Serotonin seemed to stimulate the motility of the rabbit small intestine by increasing integrated mechanical activity and tone of muscle fibers in duodenum, jejunum, and ileum. In conclusion, serotonin might alter or modulate the whole intestinal physiology. PMID- 10841432 TI - Endothelium and aortic contraction to endothelin-1 in the pregnant rat. AB - Endothelium-derived factors modulate tone and may be involved in hyporeactivity to vasoconstrictors, such as norepinephrine or angiotensin II, as has been previously described during gestation. The endothelium produces endothelin-1, a major vasoconstrictor peptide, therefore aortic contractions to endothelin-1 (10( 10) to 3 x 10(-7) M) were used to assess the role of the endothelium in pregnant Wistar rats (at 20 days of gestation). Late pregnancy is characterized by a significantly diminished systolic blood pressure in conscious rats (-17 mmHg, P < 0.001, n = 14). In pregnant and in age-matched nonpregnant female rats, endothelin-1 induced aortic contraction was greater when endothelium was present (at least P < 0.01). Indomethacin significantly reduced this contraction in aortic rings with intact endothelium in all groups. In aortic rings that had endothelium physically removed, contraction to endothelin-1 was greater in pregnant rats than in nonpregnant ones. Indomethacin decreased contraction of aortic rings in pregnant rats only. These results suggest an enhanced synthesis of vasoconstrictors by cyclooxygenases in vascular smooth muscle during pregnancy. In vessels with intact endothelium, we did not find hyporeactivity to endothelin-1 during late pregnancy. Contraction to endothelin-1 involved ET(A) receptors because it was decreased by BQ-123, an ET(A) receptor antagonist, whereas there was no significant change when using BQ-788, an ET(B) receptor antagonist. PMID- 10841433 TI - Comparison of changes evoked by GABA (gamma-aminobutyric acid) and anoxia in [K+]o, [Cl-]o, and [Na+]o in stratum pyramidale and stratum radiatum of the guinea pig hippocampus. AB - Ion-selective microelectrode recordings were made to assess a possible contribution of extracellular gamma-aminobutyric acid (GABA) accumulation to early responses evoked in the brain by anoxia and ischemia. Changes evoked by GABA or N2 in [K+]o, [Cl-]o, [Na+]o, and [TMA+]o were recorded in the cell body and dendritic regions of the stratum pyramidale (SP) and stratum radiatum (SR), respectively, of pyramidal neurons in CA1 of guinea pig hippocampal slices. Bath application of GABA (1-10 mM) for approximately 5 min evoked changes in [K+]o and [Cl-]o with respective EC50 levels of 3.8 and 4.1 mM in SP, and 4.7 and 5.6 mM in SR. In SP 5 mM GABA reversibly increased [K+]o and [Cl-]o and decreased [Na+]o; replacement of 95% O2 -5% CO2 by 95% N2 -5% CO2 for a similar period of time evoked changes which were for each ion in the same direction as those with GABA. In SR both GABA and N2 caused increases in [K+]o and decreases in [Cl-]o and [Na+]. The reduction of extracellular space, estimated from levels of [TMA+]o during exposures to GABA and N2, was 5-6% and insufficient to cause the observed changes in ion concentration. Ion changes induced by GABA and N2 were reversibly attenuated by the GABA(A) receptor antagonist bicuculline methiodide (BMI, 100 microM). GABA-evoked changes in [K+]o in SP and SR and [Cl-]o in SP were depressed by > or =90%, and of [Cl-]o in SR by 50%; N2-evoked changes in [K+]o in SP and SR were decreased by 70% and those of [Cl-]o by 50%. BMI blocked delta [Na+]o with both GABA and N2 by 20-30%. It is concluded that during early anoxia: (i) accumulation of GABA and activation of GABA(A) receptors may contribute to the ion changes and play a significant role, and (ii) responses in the dendritic (SR) regions are greater than and (or) differ from those in the somal (SP) layers. A large component of the [K+]o increase may involve a GABA-evoked Ca2+ activated gk, secondary to [Ca2+]i increase. A major part of [Cl-]o changes may arise from GABA-induced g(Cl) and glial efflux, with strong stimulation of active outward transport and anion exchange at SP, and inward Na+/K+/2Cl- co-transport at SR. Na+ influx is attributable mainly to Na+-dependent transmitter uptake, with only a small amount related to GABA(A) receptor activation. Although the release and (or) accumulation of GABA during anoxia might be viewed as potentially protectant, the ultimate role may more likely be an important contribution to toxicity and delayed neuronal death. PMID- 10841434 TI - Enhancement of hippocampal cholinergic neurotransmission through 5-HT1A receptor mediated pathways by repeated lithium treatment in rats. AB - Hippocampal cholinergic neuronal activity is reported to be regulated, at least partly, through serotonin1A (5-HT1A) receptors. Chronic lithium treatment has been shown to alter both behavioral and neurochemical responses mediated by postsynaptic 5-HT1A receptors. We investigated whether long-term lithium treatment affects central cholinergic neurotransmission through 5-HT1A receptor mediated pathways. Changes in acetylcholine (ACh) release induced by 8-hydroxy-2 (di-n-propylamino)tetralin (8-OH-DPAT), a 5-HT1A receptor agonist, in the rat hippocampus were measured using a microdialysis technique and a radioimmunoassay for ACh. Administration of lithium for 21 days resulted in a serum lithium concentration of 1.03 mM and caused little change in density or affinity of [3H]8 OH-DPAT binding sites in the hippocampus. The local application of 8-OH-DPAT into the hippocampus of lithium treated rats increased the ACh efflux in both the absence and the presence of physostigmine, a cholinesterase (ChE) inhibitor, in the perfusion fluid. The basal ACh efflux of lithium treated rats was not different from that of the control rats under normal conditions, but was significantly higher than that of the controls when ChE was inhibited. These results demonstrate that chronic lithium treatment increases spontaneous ACh release in the hippocampus under conditions of ChE inhibition, but not under normal conditions, and enhances cholinergic neurotransmission through 5-HT1A receptor-mediated pathways, and suggest that activation of 5-HT1A receptor function by lithium is related to the enhancement of hippocampal cholinergic neurotransmission. PMID- 10841435 TI - Pulmonary and systemic vascular tissue collagen, growth factor, and cytokine gene expression in the rabbit. AB - During development, the vascular wall composition of the pulmonary and systemic capacitance vessels and their intravascular pressure changes. Little is known, however, about the factors controlling vascular collagen gene expression in both circulations during growth and development. The purpose of this study was to compare the developmental changes in collagen, major growth factors, and cytokines gene expression, in order to ascertain whether a circulation specific pattern is present in the rabbit. Fetal, neonatal, and adult rabbit extrapulmonary and aortic tissues were obtained and the mRNA levels for collagen I and III, as well as major growth factors and cytokines, were measured by a semi quantitative RT-PCR technique. Collagen I, but not collagen III, expression was developmentally regulated in pulmonary vascular and aorta tissues. Collagen I expression was greatest during the fetal and neonatal period (P < 0.01) and higher in the aorta as compared with the pulmonary artery at these ages (P < 0.05). Significant developmental changes in growth factor mRNA levels were observed for TGF-beta, IGF-2, and bFGF (P < 0.01). IGF-2 mRNA levels significantly declined in both arteries from neonatal to adult, but bFGF increased only in the pulmonary artery during this transition. With regards to inducible enzymes, COX-2 mRNA levels changed developmentally, whereas iNOS mRNA levels were similar for both vessels at all ages. When comparing the two vessels, COX-2 transcripts were relatively more abundant in the adult pulmonary artery tissue and fetal aorta, with similar levels in the newborn. We conclude that circulation specific developmental regulation of collagen gene expression is present in the rabbit in a pattern that is unrelated to the intravascular pressure. PMID- 10841436 TI - Predictors of torsades de pointes in rabbit ventricles perfused with sedating and nonsedating histamine H1-receptor antagonists. AB - Several nonsedating histamine H1-receptor antagonists are associated with torsades de pointes ventricular tachycardia. The objectives of this study were to: (i) compare electrocardiographic, monophasic action potential, and arrhythmogenic effects of sedating and nonsedating H1-receptor antagonists, and (ii) identify correlates of drug-induced torsades de pointes in an isolated ventricle model. Isolated, electrically paced (1-3 Hz) rabbit ventricles were Langendorff-perfused with either drug-free Tyrode's solution or one of the following: (i) the sedating H1-receptor antagonist hydroxyzine (0.1-30 microM), (ii) cetirizine, a nonsedating metabolite of hydroxyzine (1-300 microM), and (iii) the nonsedating, putatively arrhythmogenic H1-receptor antagonist astemizole (0.1-30 microM). Volume conducted electrocardiographic signals and monophasic action potentials from the periapical left ventricular endocardium and epicardium were recorded. There were no apparent changes in control (n = 15) or hydroxyzine-perfused (n = 7) hearts. Cetirizine (n = 13) produced a mild biphasic electrocardiographic QT interval prolongation and was associated with early afterdepolarizations, but not with torsades de pointes. Astemizole (n = 11) lengthened QT intervals, and at high concentration (30 microM) induced torsades de pointes in 10 of 11 hearts (P < 0.001 vs. all other groups). These findings are consistent with previously reported repolarizing current inhibition by cetirizine, but may additionally indicate "compensatory" inhibition of inward currents at higher concentrations. By contrast, astemizole-induced changes are consistent with unopposed repolarizing current inhibition. PMID- 10841437 TI - Acute effects of methoxamine on left ventricular-arterial coupling in streptozotocin-diabetic rats: a pressure-volume analysis. AB - We determined the acute effects of methoxamine, a specific alpha1-selective adrenoceptor agonist, on the left ventricular-arterial coupling in streptozotocin (STZ)-diabetic rats, using the end-systolic pressure-stroke volume relationships. Rats given STZ 65 mg x kg(-1) iv (n = 8) were compared with untreated age-matched controls (n = 8). A high-fidelity pressure sensor and an electromagnetic flow probe measured left ventricular (LV) pressure and ascending aortic flow, respectively. Both LV end-systolic elastance E(LV,ES) and effective arterial elastance Ea were estimated from the pressure-ejected volume loop. The optimal afterload Q(load) determined by the ratio of Ea to E(LV,ES) was used to measure the optimality of energy transmission from the left ventricle to the arterial system. In comparison with controls, diabetic rats had decreased LV end-systolic elastance E(LV,ES), at 513 +/- 30 vs. 613 +/- 29 mmHg x mL(-1), decreased effective arterial elastance Ea, at 296 +/- 20 vs. 572 +/- 48 mmHg x mL(-1), and decreased optimal afterload Q(load), at 0.938 +/- 0.007 vs. 0.985 +/- 0.009. Methoxamine administration to STZ-diabetic rats significantly increased LV end systolic elastance E(LV,ES), from 513 +/- 30 to 602 +/- 38 mmHg x mL(-1), and effective arterial elastance Ea, from 296 +/- 20 to 371 +/- 28 mmHg x mL(-1), but did not change optimal afterload Q(load). We conclude that diabetes worsens not only the contractile function of the left ventricle, but also the matching condition for the left ventricular-arterial coupling. In STZ-diabetic rats, administration of methoxamine improves the contractile status of the ventricle and arteries, but not the optimality of energy transmission from the left ventricle to the arterial system. PMID- 10841438 TI - Effect of nitric oxide synthase inhibition on cardiovascular and hormonal regulation during pregnancy in the rat. AB - Recent studies have shown that nitric oxide (NO) biosynthesis increases in pregnancy and that inhibition of nitric oxide synthase (NOS) induces some pathological processes characteristic of preeclampsia. The current project sought to study the effect of the NOS inhibitor Nomega-nitro-L-arginine methyl ester (L NAME, 10 microg x min(-1), sc for 7 days) on plasma volume, plasma atrial natriuretic factor (ANF), plasma endothelin-1 (ET), and plasma renin activity (PRA) during gestation in conscious rats. NOS inhibition caused mean arterial pressure to increase in both virgin and 21-day pregnant rats. Plasma volume fell in the pregnant rats [L-NAME, 4.5 +/- 0.3 mL x 100 g(-1) body wt. (n = 7) vs. D NAME, 6.8 +/- 0.2 mL x 100 g(-1) body wt. (n = 10); P < 0.05] but not in the virgin rats [L-NAME, 4.3 +/- 0.1 mL x 100 g(-1) body wt. (n = 6) vs. D-NAME, 4.8 +/- 0.2 mL x 100 g(-1) body wt. (n = 8)]. There was no effect of NOS inhibition on plasma ANF levels or PRA in either the virgin or pregnant rats. However, L NAME did decrease plasma ET levels in the pregnant rats [L-NAME, 19.6 +/- 1.6 pg x mL(-1) (n = 8) vs. D-NAME, 11.6 +/- 2.5 pg x mL(-1) (n = 9); P < 0.05]. Our results confirm that NO is involved in cardiovascular homeostasis in pregnancy; NOS inhibition selectively reduces plasma volume in pregnant rats, thus mimicking a major pathophysiological perturbation of preeclampsia. However, it does not induce the hormonal changes characteristic of preeclampsia, namely the decrease in PRA and increase in plasma ET and ANF levels. PMID- 10841439 TI - Renal adenosine A3 receptors in the rat: assessment of functional role. AB - The functional roles of adenosine A3 receptors in the rat kidney were assessed for the first time with respect to A1 receptor-mediated responses. Utilizing a chronically instrumented conscious rat preparation, we tested renal excretory responses to acute administration of the A3 receptor antagonists 3-ethyl-5-benzyl 2-methyl-6-phenyl-4-phenylethynyl-1 ,4-(+)-dihydropridine-3,5-dicarboxylate (MRS 1191) and 9-chloro-2-(2-furyl)-5-phenylacetylamino-[1,2,4]-triazolo[1,5-c]qu inazoline (MRS-1220) with reference to the effects of the A1 receptor antagonist 1,3-dipropyl-8-cyclopentylxanthine (DPCPX). The intravenous administration of DPCPX resulted in significant increases in fluid and sodium excretions without affecting glomerular filtration rate (GFR). This suggests that DPCPX-induced diuretic and natriuretic responses are related to decreased tubular reabsorption. However, neither MRS-1191 nor MRS-1220 alone affected fluid or sodium excretions, or GFR, indicating lack of an effect of either compound on renal function. On the other hand, the co-administration of MRS-1220 with DPCPX abolished both the diuretic and natriuretic responses to DPCPX, being suggestive of antagonism between these two compounds. MRS-1191, however, did not affect the DPCPX-induced fluid and sodium excretions. Neither the A1 nor the A3 receptor antagonists altered potassium excretion individually or in combination. The data suggest that while adenosine A1 receptors are involved in the regulation of renal fluid and sodium transport, A3 receptors do not appear to have a major role in regulation of renal excretory function under baseline physiological conditions. PMID- 10841440 TI - Fine structure, synaptology and immunocytochemistry of large neurons in the rat dorsal cochlear nucleus connected to the inferior colliculus. AB - Neurons in the rat dorsal cochlear nucleus that project to the inferior colliculus (pyramidal and giant) were retrograde labelled with wheat germ agglutinin conjugated to horseradish peroxydase. Both cell types showed a similar ultrastructural feature, particularly the rough endoplasmic reticulum was well developed and sometimes surrounded the nucleus. The synaptological profile was similar in pyramidal and giant cells. Axo-somatic terminals covered 40-70% of the perimeter of pyramidal cells and 35-60% of the perimeter of giant neurons. Giant neurons featured bipolar or multipolar shape and different orientation but they possessed a similar synaptic profile. Most axo-somatic terminals contained flat and pleomorphic synaptic vesicles, some pleomorphic vesicles. Few terminals contained round vesicles. These cells were consistently immuno-negative for both glycine and GABA and variably positive for glutamate. The immunoelectron microcopic study of thin sections showed that glycine immunoreactivity was constantly present in terminals enriched with flat vesicles, which often did not show GABA immunoreactivity. Few anterograde labelled boutons containing flat vesicles were in contact with the proximal dendrites and the cell bodies of pyramidal and giant neurons. The origin of these terminals is discussed. No other cells of the dorsal cochlear nucleus, in particular cartwheel and tuberculo ventral neurons, were in contact with labelled boutons. The present results suggest that descending inhibitory collicular projections are essentially directed to the large excitatory neurons of the dorsal cochlear nucleus. PMID- 10841441 TI - Ultrastructural changes in the sinuatrial and atrioventricular nodes of the heart of the monkey (Macaca fascicularis) after bilateral vagotomy. AB - This study describes the ultrastructural changes in the sinu-atrial (SA) and atrio-ventricular (AV) nodes of the monkey (Macaca fascicularis) after bilateral mid-cervical vagotomy at 1, 3, 5, 7 and 14 days post-operations. The changes were similar in both types of nodal cells. The most obvious feature of the degenerating nodal cells was the swollen mitochondria with disrupted cristae. Other changes include increased granular sarcoplasmic reticulum, increased glycogen particles, vacuolation of mitochondria and increased lysosomal activity. Axonal profiles in the vicinity of the nodal cells showed swelling and vacuolation. Cardiac neurons also showed some changes such as distended granular endoplasmic reticulum, increased accumulation of glycogen particles and increased lipofuscin granules. Macrophages and Schwann cells were the main scavengers in removing the degenerated nodal cells and axonal profiles. In the case of affected cardiac neurons, satellite cells seemed to act as main scavenger cells. It is postulated that the nodal cells are dependent on the incoming fibres of the vagus nerve for their survival. By an understanding of the ultrastructural changes in the nodal cells after bilateral vagotomy, it may help in developing new strategies to explore in depth of the conducting system of the heart. PMID- 10841442 TI - Disruption of patterns of immunoreactive glial fibrillary acidic protein processes in the Cebus Apella striate cortex following loss of visual input. AB - Long, interlaminar, astroglial processes and its patterned organization in the striate cortex of adult primates was previously described. Loss of visual input following bilateral retinal detachment and degeneration in an adult Cebus apella monkey resulted three months later in reduction of interlaminar processes immunoreactive to Glial Fibrillary Acid Protein antibody, loss of the honeycomb like pattern normally present in tangential sections, and loss of high density patches of terminal segments of those processes in the opercular striate. These results further indicate the highly interactive nature of neuron-glial cerebral cortex architecture, and the dynamic regulation of astroglial interlaminar processes. PMID- 10841443 TI - Electron microscopic studies on NO-synthase activity in brain phagocytes of rat cerebral cortex after ischemic and traumatic brain injury. AB - In the present study we used a cytochemical method for electron microscopy for nitric oxide synthase (NOS) detection in rat cerebral cortex. The animals were subjected to total brain ischemia and mechanical trauma. Tissue was fixed in 3% paraformaldehyde and 0.5% glutaraldehyde which enabled excellent preservation of cellular ultrastructure and resulted in an increase in method reproducibility. The results show the distribution of the particles reflecting the sites of NOS activity in the capillaries and other, more distant locations. NOS activity was found in thrombocytes, endothelium and pericytes. Moreover, the morphologically differentiated perivascular phagocytes demonstrated NOS reaction product in their cytoplasm. These phagocytes containing polymorphic phagolysosomes, surrounded the fragments of damaged neurons. We propose that these cells are descendants of blood-borne monocytes and transformed pericytes. In the areas distant to the sites of injury we noticed cells showing ultrastructural features of activated microglia. These cells were in a close contact with neural perikarya and contained iNOS-specific reaction product in the cytoplasm. In conclusion, our studies revealed differentiated forms of brain phagocytes, the occurrence of which depended on the type of cerebral injury. These cells demonstrated NOS reaction product suggesting their role in cerebral NO production during ischemia and other forms of brain injury. PMID- 10841444 TI - Validation of a four-vessel occlusion model for transient global cerebral ischemia in dogs. AB - A new model of transient global cerebral ischemia in dogs with minimal measures of intervention is described together with a simple scale for evaluation of functional outcome. During pentobarbital anesthesia, a global cerebral ischemia lasting seven minutes was induced by a four-vessel occlusion and a controlled systemic hypotension. The reperfusion phase begun after removal of arterial clamps, and the animals were sacrificed by perfusion fixation 24 hours latter. The efficiency of controlled systemic hypotension in diminishing collateral blood flow was validated in two experimental groups with different cerebral filling pressure (CFP). Severe ischemia group (CFP 1.0-1.5 kPa) underwent near-complete ischemia as indicated by rCBF, electroencephalography, and histologically documented ischemic neuronal changes. Mild ischemia group (CFP 2.5-3 kPa) animals experienced reduction in cerebral blood flow well above the ischemic threshold, had better functional outcome as well as no ischemic neuronal changes on light microscopy. This model consistently produces global cerebral ischemia in dogs with minimal surgical intervention and pharmacological support, and without intracranial hypertension, cardiac arrest or asphyxia. We recommend this model for outcome-oriented studies of complete forebrain ischemia in dogs. PMID- 10841445 TI - Cell-cell coupling in cultures of striatal and cortical astrocytes of the monkey Cebus apella. AB - Astrocytes were cultured from striatum and neocortex of fetal (embryonic day 90) monkeys (Cebus apella). The cultures grew well, and the cells retained viability after freeze-storage and thawing. The cells displayed depolarized membrane potentials (-19 and -33 mV, for striatal and cortical cells, respectively) and the vast majority of cells were dye-coupled to a mean of 7 (1-18) neighbouring cells. Cell coupling was blocked by octanol (0.25 and 0.5 mM) but was independent of high K+ (10 and 50 mM) and glutamate (500 microm). Thus, cultures of fetal primate astrocytic cells are established as a model system for studies on astroglial cell-cell coupling. PMID- 10841446 TI - Lectin binding sites in the vomeronasal organ and the olfactory epithelium of the tree shrew tupaia belangeri. AB - Lectin binding histochemistry was performed on the peripheral parts of the vomeronasal and olfactory system of Tupaia belangeri to investigate the distribution and density of defined carbohydrate terminals on the cell surface glycoproteins of the specific receptors, nerves and associated glands. Dolichus biflorus lectin staining was identifiable in all cells of the sensory epithelia of both systems, with the exception of the olfactory basal cells. The vomeronasal nerve expressed alpha-N-acetylgalactosamine, the olfactory nerve did not. Differences in the Dolichus biflorus lectin staining pattern were noted in the sensory and the non-sensory epithelium of the vomeronasal organ of pregnant and non-pregnant Tupaia belangeri. Like in other mammals examined so far, the systems were characterized by a moderate to high alpha-fucose density, only the supporting cells of the olfactory epithelium remaining unstained by Ulex europaeus I lectin. Bandeiraea simplicifolia II lectin binding was moderate in the olfactory glands and the brush border. In the vomeronasal organ, sugar residues of alpha- and beta-N-acetyl-D-glucosamine were found selectively in the transition zone between the two vomeronasal epithelia, where regeneration should take place. We tentatively conclude that specific glycoproteins, whose terminal sugars were detected by lectin binding, might be related to the chemoreception and transduction of the olfactory/pheromonal message into a nervous signal or to the histogenesis of the olfactory/vomeronasal system. This study showed that beside the species, pregnancy might play a role in these processes in the vomeronasal organ. PMID- 10841447 TI - The brain lesion influences the numbers of peripheral blood leukocytes in the rat. AB - The mutual relationship between the central nervous and immune system are intensively studied. The lesion of distinct structures of the rat brain such as septum influence the model immune response such as lymphocyte proliferation and delayed skin hypersensitivity. Employing the model of the damage of septum in the rat brain by electrolesion we demonstrated the decrease of the number of peripheral blood leukocytes, mainly cells exhibiting CD25 and CD45RA antigens in the rat. The striatum destruction has much lower influence on the studied parameters, which suggests a specific effect of the septum on these hematological parameters. PMID- 10841448 TI - Benzodiazepine binding sites in the human hypothalamus. Autoradiographic study. AB - Using in vitro labelling and autoradiographic techniques, we have analyzed the fine and the detailed distribution of benzodiazepine binding sites in the post mortem human hypothalamus. Binding sites were labelled in mounted tissue sections from adult brains, using the selective high affinity ligand [3H]-Flunitrazepam. A heterogeneous distribution of benzodiazepine binding sites was found throughout the rostrocaudal extent of human hypothalamus. The autoradiographic labelling was shown in the three hypothalamic parts, i.e., anterior, mediobasal and posterior levels. At the anterior level, the highest densities were present in the diagonal band of Broca, the preoptic area (medial and lateral parts) and the septohypothalamic nucleus. At the mediobasal hypothalamic level, the highest densities were mainly localized in the ventromedial nucleus, whereas the other structures were moderately labelled with [3H]-Flunitrazepam. The mammillary complex as well as the posterior hypothalamic area represented the most heavily labelled structures in the posterior hypothalamus. The results obtained in this study, indicate the presence of a large and heterogeneous distribution of benzodiazepine binding sites in human adult hypothalamus. This could support their implication in the control of distinct neural functions (like neuroendocrine role). PMID- 10841449 TI - Ultrastructural study of the effects of 6-hydroxydopamine on the sinuatrial and atrioventricular nodes of the heart of the monkey (Macaca fascicularis). AB - The present study examined the ultrastructural changes in the sinuatrial and atrioventricular nodes of the heart of the monkey (Macaca fascicularis) after 6 hydroxydopamine (6-OHDA) at survival times of 1, 3, 5, 7 and 14 days. Changes ranged from dissolution of the cytoplasm to amorphous appearance and darkening of the nodal cells. Initially, the ultrastructural changes were quite similar in both sinuatrial (SA) and atrioventricular (AV) nodal cells but in the later stage, especially at fourteen days, affected SA nodal cells showed empty-looking appearance while affected AV nodal cells displayed a darkened appearance. The cardiac neurons also showed ultrastructural changes such as diffuse accumulation of glycogen particles, distended cisternae of the rough endoplasmic reticulum and increased lipofuscin granules. Some of the vacuolated axonal profiles containing large dense-cored vesicles were in close association with the somata of the cardiac neurons. There were also changes in the non-neuronal cells such as darkening and vacuolation of the cells capping the neurons. Macrophages and Schwann cells were activated to engulf the degenerating nodal cells and axonal profiles. The ultrastructural changes in the nodal cells and the cardiac neurons reflect a disturbance in the cell metabolism presumably brought about by the impairment of the sympathetic nervous system. PMID- 10841450 TI - NADPH-diaphorase expression in the hypothalamo-hypophysial system of different catfish. AB - The distribution of NADPH-d activity was studied in the hypothalamus and in the pituitary gland of 15 species of catfish. Seven hypothalamic nuclei, four fiber bundles, as well as cells located in the adenohypophysis were labeled by NADPH-d histochemistry. Reactive somata were found in the nucleus praeopticus periventricularis, the paraventricular division of the nucleus praeopticus, the supraoptic division of the nucleus praeopticus, the nucleus lateralis tuberis, the paraventricular organ, the nucleus recessus lateralis, the nucleus recessus posterioris, and in the adenohypophysis. In some species, an inconsistent number of these structures lacked NADPH-d activity. These results are compatible with the notion that NADPH-d activity expressing cells in the hypothalamus and in the pituitary are involved in the control of hormone regulation. PMID- 10841451 TI - PCNA positivity in the telencephalic matrix areas in the adult of a newt, Triturus carnifex. AB - The Proliferating Cell Nuclear Antigen (PCNA) is an auxiliary protein of the DNA polymerase delta, belonging to the cyclin family, which attains appreciable levels only in those phases of the cell cycle in which DNA synthesis occurs. Using an immunocytochemical method that exploits this proliferative marker, we observed a certain PCNA positivity in the telencephalon of normal adult individuals of Triturus carnifex. The elements that display this peculiarity occupy an ependymal and/or sub-ependymal site. They are located in the anterior portion of the telencephalon only at the dorsal level and tend to decrease in number until they disappear temporarily as the intermediate portion is approached. In the posterior portion of the telencephalon, the dorsal labelling patterns, the size of which is however smaller than that observed more anteriorly, reappear and in the ventral region a large population of labelled cells appears. Then, again proceeding in a caudal direction, while the PCNA positive elements lying dorsally diminish and disappear, they persist in the ventral region and gradually taper off until they final disappear where the telencephalic ventricles come together. This immunocytochemical picture supports the findings of other authors in adult Urodeles under both normal and experimental conditions (with classical and autoradiographic histological techniques) concerning the persistence of the telencephalon in dorsal and ventral germinative areas responsible for physiological and plastic neurogenetic events. PMID- 10841452 TI - 5-HT potentiates GABA- and glycine-activated chloride currents on the same neurons in rat spinal cord. AB - The electrophysiological and pharmacological properties of GABA- and glycine (Gly)-induced responses were investigated in neurons acutely dissociated from the rat sacral dorsal commissural nucleus (SDCN) using the nystatin perforated patch recording configuration under voltage-clamp conditions. At a holding potential of -40 mV, the currents evoked by GABA and Gly increased in a sigmoidal fashion with increasing agonist concentration. The reversal potentials of I(GABA)and I(Gly) were close to the Cl- equilibrium potential. Bicuculline (BIC) and strychnine (STR) suppressed I(GABA) and I(Gly) in a concentration-dependent manner, although they have different affinities for GABA and Gly. 5-HT potentiated both I(GABA) and I(Gly) via intracellular protein kinase C on the same neurons. The results indicated that the acutely dissociated SDCN neurons responded to both exogenous GABA and Gly, which activated GABA(A) and STR-sensitive Gly receptors, respectively, and 5-HT may produce spinal antinociception through postsynaptic potentiation of I(GABA) and I(Gly) when they are coreleased; GABA and Gly may act as cotransmitters implicated in the control of spinal nociceptive signal processing in the mammalian spinal cord. PMID- 10841453 TI - The activity of a transient potassium current in retinal glial (Muller) cells depends on extracellular calcium. AB - The modulating effects of varying extracellular concentrations of Ca2+ ([Ca2+]e) and of other divalent cations on the fast transient (A-type) K+ current (I(A)) of freshly isolated Muller glial cells from rabbit and human retinae were studied with the whole-cell patch-clamp method. The I(A) of Miller cells was voltage independently blocked by extracellular 4-aminopyridine (4AP) with a 50 % reduction achieved at 0.94 mM 4AP. The I(A) amplitude was elevated by increased extracellular [K+]. Elevation of the [Ca2+]e had three effects on the glial I(A): (i) it concentration-dependently shifted both the activation and inactivation curves towards less negative membrane potentials, (ii) it increased the peak current amplitude, and (iii) it slowed down the activation and inactivation kinetics. Particularly at depolarized membrane potentials, the I(A) was enlarged and broadened when the [Ca2+]e was increased. Various divalent cations also exerted these effects, although at different concentrations. While Zn2+, Cd2+, Cu2+ and Pb2+ modulated the I(A) in the micromolar range, Mg2+ and Ba2+ had effects in the millimolar range. Extracellular acidification produced a positive shift in the voltage dependence of I(A) gating. However, alterations of the extracellular pH did not abolish the Ca2+ effects on I(A); this indicates that protons and Ca2+ ions mediate their effects on glial K(A) channels by different mechanisms or binding sites, respectively. Physiological (i.e., activity dependent) changes of the extracellular concentration of divalent cations and of the extracellular pH should influence the retinal excitability via modulation of glial K+ currents. The activation of glial I(A) by divalent cations at depolarized voltages supports a repolarization and, therefore, the maintainance of a hyperpolarized glial membrane potential during periods of increased neuronal activity. PMID- 10841454 TI - A light and electron microscopic study of GAT-1 positive cells in the monkey brainstem and spinal cord. AB - The distribution of the GABA transporter GAT-1 was studied in the monkey brainstem and spinal cord, using an affinity purified polyclonal antibody against a synthetic peptide corresponding to the C terminus of GAT-1. Very dense staining was observed in the interpeduncular nucleus, the inferior olivary nucleus and the substantia gelatinosa of the spinal cord, whilst dense labelling was observed in the substantia nigra, cochlear nuclei, vestibular nuclei, the spinal nucleus of V, the area postrema and the ventral horn of the spinal cord. Electron microscopy showed that the labelled profiles consisted of axon terminals that formed symmetrical synapses, consistent with GABAergic terminals. Many of the nuclei that were densely labelled for GAT-1 were those that received primary auditory, vestibular, or somatosensory inputs and the high density of GAT-1 in these nuclei suggests that GAT-1 plays an important role in terminating the inhibitory effects of GABA, at these nuclei. PMID- 10841455 TI - Ultrastructural changes in the atrioventricular valves of streptozotocin-induced diabetic rats. AB - The present study describes ultrastructural changes in the atrioventricular (AV) valves of diabetic rats at 3, 6, 9 and 12 months. At 3 and 6 months of diabetes, the interstitial cells were characterized by an accumulation of vacuoles, lysosomes, electron-dense vesicles, mitochondria and cisternae of rough endoplasmic reticulum in the cytoplasm. There were numerous collagen fibres in the interstitial space. Unmyelinated axons were ensheathed by Schwann cells. Infiltration of macrophages was observed near the interstitial cells. Each macrophage showed a large round or oval nucleus containing heterochromatin masses at the periphery of the cell nucleus. At 9 and 12 months of diabetes, the interstitial cells contained numerous vacuoles, dilated mitochondria, agranular vesicles and a prominent multivesicular body in the cytoplasm. Degenerating unmyelinated nerve fibres were encountered near the interstitial cells. Phagocytic macrophages contained numerous vacuoles of various sizes, which occupied most of the cytoplasmic area. Several vacuoles and degenerated electron dense granules (some of them appeared to be fragmented) were present in the cytoplasm of interstitial cells and macrophages. It is concluded that interstitial cells in the AV valves contribute to valvular dysfunction in the streptozotocin-induced diabetic rats. PMID- 10841456 TI - Infraorbital nerve transection increases NADPH-diaphorase activity in the rat mesencephalic trigeminal nucleus. AB - Complete information concerning possible alterations in nitric oxide production in the rat brainstem trigeminal system after peripheral nerve lesion is still lacking. This being the case, density of NADPH-diaphorase positive neurons in the trigeminal mesencephalic nucleus after experimental infraorbital nerve transection was studied. In the lesioned side, an ipsilateral increase in NADPH-d positive neurons was found at postoperative days 4 and 6 with respect to contralateral, without changes after a 2 months period. These data suggest that nitric oxide could be involved in regeneration of afferent fibers concerned with the periodontal receptors of maxillary teeth. PMID- 10841457 TI - Non-specific immunity and ketone bodies. II: In vitro studies on adherence and superoxide anion production in ovine neutrophils. AB - The effects of the ketone bodies beta-OH-butyrate and acetoacetate (2.4 or 4.8 mmol/l), administered singly or simultaneously in vitro, on adherence and superoxide anion (SO) production in ovine neutrophils were investigated by simultaneous assay in 96-well microplates. Because the acetoacetate used was a lithium salt, the effect of 2.4 and 4.8 mmol/l lithium chloride was also tested. Neutrophils from eight non-lactating, non-pregnant ewes were used. SO release from neutrophils was found to be very low in basal conditions and was apparently not stimulated by contact with plastic. Administration of 10(-7) mol/l phorbol myristate acetate (PMA) caused a rapid increase and release of SO production, but smaller than that induced by co-stimulation with plastic and 10(-7) mol/l PMA. LiCl (2.4 and 4.8 mmol/l) significantly increased PMA-stimulated release, but inhibited plastic and PMA co-stimulated SO release. Administration of 2.4 mmol/l ketone bodies inhibited plastic and PMA-costimulated SO release, but the effect of acetoacetate could be due to the lithium component. Administration of 4.8 mmol/l ketone bodies had no effect. Adherence was significantly increased by contact with plastic, and moreover by 10(-7) mol/l PMA. The effect was similar when PMA was acting alone or with plastic. Neither basal nor stimulated adherence were affected by 2.4 or 4.8 mmol/l ketone bodies. LiCl at a concentration of 4.8 mmol/l increased PMA and plastic co-stimulated adherence. The results suggest that, in sheep, only the ketone body beta-OH butyrate at concentrations seen in mild ketosis, could decrease bactericidal activity, while adherence is not affected. In addition to other factors that could impair the efficiency of the immune system in ketotic ruminants, the reduced bactericidal activity may contribute to the higher occurrence of infectious disease in these animals. PMID- 10841458 TI - Suppression of feed intake after parenteral administration of D-beta hydroxybutyrate in pygmy goats. AB - The effect of intraperitoneal injection of D,L- or D-beta-hydroxybutyrate on feed intake and plasma metabolites was investigated in pygmy goats. The combined intraperitoneal injection of D,L-beta-hydroxybutyrate or D-beta-hydroxybutyrate (15 mmol/kg0.75) and 1,3-butanediol (6.6 mmol/kg0.75), a ketogenic substrate, decreased cumulative food intake while the same dose of 1,3-butanediol or DL-beta hydroxybutyrate alone had no effect. The decrease in food intake after combined injection of D-beta-hydroxybutyrate and 1,3-butanediol was characterized by a significant decrease in meal frequency and a prolongation of the latency to eat. The hypophagic effect of the combined injection of D-beta-hydroxybutyrate and 1,3 butanediol was significant for 8 h, while the hypophagia after D,L-beta hydroxybutyrate and 1,3-butanediol was significant for only 2 h after injection. Injection of D,L-beta-hydroxybutyrate increased plasma D-beta-hydroxybutyrate levels to 0.55 mmol/l and decreased plasma free fatty acids. Addition of 1,3 butanediol (6.6 mmol/kg0.75) to the injection increased plasma D-beta hydroxybutyrate level up to 0.75 mmol/l. The results show that parenteral administration of D-hydroxybutyrate reduces feed intake in pygmy goats and that the hypophagia appears to be related to the amount of D-beta-hydroxybutyrate injected. The hypophagia seems to be related to elevated plasma D-beta hydroxybutyrate concentration, and the threshold concentration appeared to be about 0.7 mmol/l under the experimental conditions of this study. PMID- 10841459 TI - Fructo-oligosaccharide supplementation: effects on metabolic, endocrine and hematological traits in veal calves. AB - Fructo-oligosaccharides (FOS) are soluble fibres which exert various effects in the gastrointestinal tract, and induce metabolic and endocrine changes. The effects are favourable in diabetes mellitus, and may be favourable in veal calves, which during late periods of fattening often develop hyperglycemia, glucosuria and insulin resistance, especially during high lactose intake. Based on this we have studied metabolic, endocrine and haematological traits in veal calves (Simmental x Red Holstein) fed FOS (10 g/day; group GrF) or no FOS (group GrC). Whole milk and milk replacer in both groups, on a kg body weight basis, were fed in identical amounts. Experiments, lasting for 3 weeks, started when calves were 10 weeks old and weighed 117 kg. During week 3 lactose was supplemented to enhance post-absorptive glucose loads. Feed intakes were similar in both groups, but weight gain tended to be higher in GrF than GrC. The post prandial increase of glucose concentrations was significantly smaller, of lactate tended to be smaller, and growth hormone peak frequency tended to be lower, whereas maximal insulin concentrations reached post-prandially were significantly higher in GrF than GrC. Eosinophil granulocytes increased during FOS feeding. In conclusion, FOS had basically similar effects on metabolic and endocrine traits in veal calves as in animals and humans with diabetes mellitus, but changes were small, albeit more prominent after lactose loads. PMID- 10841460 TI - Association analysis between bilateral convergent strabismus with exophthalmus and milk production traits in dairy cattle. AB - Bilateral convergent strabismus with exophthalmus (BCSE) is characterized in cattle by a symmetrical antero-medial rotation and protrusion of the eyeballs. This eye defect is caused by an inherited, centrally insufficient function of the eye muscles recti laterales and retractores. In German Brown cattle a monogenic, autosomal dominant inheritance proved to be most probable in complex segregation analysis. In our study running between October 1993 and May 1995 a total of 200 affected German Brown cows was studied. The investigation of the association between milk production traits and BCSE was based on 10,960 German Brown cows. The analysis revealed no significant differences between affected and non affected cows, nor between cow families with and without affected members. There was no indication of an association between milk production traits and occurrence of BCSE within cow families. Linkage or pleiotropy of the BCSE locus with quantitative trait loci for milk production traits may be rather unlikely. PMID- 10841461 TI - Pharmacokinetics of miocamycin following intravenous administration to cattle. AB - The plasma pharmacokinetics for a single intravenous dose (10 mg/kg body weight) of miocamycin (a 16-membered macrolide drug) was investigated in Holando Argentino cattle (n = 5). Blood drug concentrations were determined by a microbiological method and data were best-fitted to a two-compartment open model. The pharmacokinetic profile consisted of a short distribution phase (t1/2 alpha = 7.41 +/- 0.53 min), followed by an extended terminal elimination phase (t1/2 beta = 2.49 +/- 0.23 h). The volume of distribution at steady-state was large (2.13 +/ 0.17 l/kg), suggesting extensive tissue distribution, the clearance value was 0.60 +/- 0.03 l/h. PMID- 10841463 TI - Intramolecular AB - 2-Propynyldiarylacetylenes undergo thermal intramolecular [4 + 2] cycloaddition to give benzo[b]fluorene derivatives in good yields. The hybridization of the tether connecting the reacting alkynes has a pronounced effect on the course of the reaction. Theoretical calculations and isotopic labeling studies support a mechanism which involves the generation of a cyclic allene intermediate that evolves to the final benzo[b]fluorene. PMID- 10841462 TI - Twenty-four-hour patterns of hormones and metabolites in week 9 and 19 of lactation in high-yielding dairy cows fed triglycerides and free fatty acids. AB - Effects of feeding crystalline triglycerides (TGA), free fatty acids (FFA), and a starch-rich ration (STA) on metabolite and hormone concentrations in blood plasma were studied in high-yielding dairy cows over a 24-h period in week 9 and 19 of lactating. Energy-corrected milk production in the three groups was similar, but was lower in week 19 than in week 9. Energy and protein intakes were greater in week 9 than in week 19, but energy and protein balances in the three groups and in weeks 9 and 19 were similar. Plasma glucose and triglyceride concentrations were lower in week 9 than in week 19. In cows fed FFA, glucose concentrations were highest in week 9. Plasma triglyceride, phospholipid and cholesterol concentrations were highest, whereas beta-hydroxybutyrate concentrations were lowest in FFA-fed cows in weeks 9 and 19. Concentrations of insulin-like growth factor-I in week 19 were lower in cows fed TGA and FFA than in those fed the starch-rich ration. Post-prandial responses were usually greater following morning than afternoon meals. Fructosamine, albumin, urea, growth hormone, thyroxine, and 3,5,3'-triiodothyronine concentrations were similar in weeks 9 and 19 and were not influenced by dietary treatment or feeding times. In conclusion, there were distinct metabolic and endocrine effects of feeding TGA and FFA compared with STA and the concentrations as well as the 24-h changes of various metabolic and endocrine traits in weeks 9 and 19 of lactation were also different. PMID- 10841464 TI - A valine-derived lithiated 3-methylthiomethyl-1,3-oxazolidin-2-one for enantioselective nucleophilic hydroxymethylation, formylation, and alkoxycarbonylation of aldehydes AB - The 3-methylthiomethyl-4-isopropyl-5,5-diphenyl-1,3-oxazolidin-2-one (I, prepared in three steps from Boc-valine ester) is lithiated and added to aldehydes, with protecting in situ trapping of the primary adducts, to give the N,S-acetal derivatives II of 2-hydroxy aldehydes in high yields and diastereoselectivities. Cleavage (with ready recovery of the oxazolidinone auxiliary) is possible, to afford, for instance, enantiopure 1,2-diols, selectively protected (OBn, OMOM, OTBS) in the 2-position. PMID- 10841465 TI - A glycosylation protocol based on activation of glycosyl 2-pyridyl sulfones with samarium triflate. AB - [reaction--see text] Reaction of glycosyl 2-pridyl sulfones (e.g.,2) with alcohols and samarium(III) triflate affords glycosides in moderate to excellent yields. Benzylated sulfones can be activated in preference to their benzoylated counterparts, and the methodology has been used to prepare di- and trisaccharides containing both furanose and pyranose residues. Thioglycosides do not react under these conditions, and the sulfones are inert to the N-iodosuccinimide/silver triflate promoter system commonly used to activate thioglycosides. This selectivity allowed the efficient preparation of oligosaccharides via orthogonal glycosylation protocols. PMID- 10841466 TI - Chiral action at a distance: remote substituent effects on the optical activity of calyculins A and B. AB - [structure--see text] Calyculins A and B differ only by the (E)- vs (Z) configuration at C(2). Yet, they show a large difference in optical rotations. We demonstrate a new strategy that provides a physical analysis of this long-range chiro-optical effect by Boltzmann-averaged atomic contribution mapping. The polarizability characteristics of the CN substituent rather than the perturbation of the stereogenic centers or the introduction of asymmetry into the polyene chain give rise to the remarkable difference in rotation angles. PMID- 10841468 TI - Improvement in olefin metathesis using a new generation of ruthenium catalyst bearing an imidazolylidene ligand: synthesis of heterocycles AB - A number of heterocycles have been prepared in very good yields using 1,3 dimesitylimidazol-2-ylidene ruthenium benzylidene 1. This catalyst displays increased activity for ring-closing metathesis of some hindered heterodienes which did not cyclize using the Grubbs catalyst 2. The scope of the olefin metathesis has been expanded. PMID- 10841467 TI - Studies in marine polypropionate synthesis: total synthesis of (-)-baconipyrone C. AB - [reaction--see text] An asymmetric total synthesis of the unusual siphonariid metabolite, (-)-baconipyrone C (3), is described. Key steps included a tin(II) mediated aldol coupling for the preparation of the carboxylic acid 17 and two different boron-mediated aldol additions leading to alcohol 8. Ester formation using modified Yamaguchi conditions gave 24, leading on PMB deprotection to (-) baconipyrone C. PMID- 10841469 TI - Addition of benzylzinc halides to alkenyl(phenyl)iodonium triflates: stereoselective synthesis of trisubstituted alkenes AB - Benzylic organozinc reagents generated by insertion of zinc metal into benzyl bromine bonds react with alkenyl(phenyl)iodonium triflates to provide single stereoisomers of trisubstituted olefins. The extremely high reactivity of the phenyliodonio moiety allows these reactions to be performed in the absence of copper salts or palladium catalysts. The reaction is performed from -40 degrees C to room temperature in THF. Excellent yields of the desired cross-coupled products have been obtained despite the occurrence of a competing electron transfer-induced fragmentation. PMID- 10841470 TI - Conformational ordering of apolar, chiral m-phenylene ethynylene oligomers AB - A series of m-phenylene ethynylene oligomers containing nonpolar, (S)-3,7 dimethyl-1-octanoxy side chains have been synthesized and studied. In apolar alkane solvents, oligomers of sufficient length (n > 10) were found to adopt a helical conformation with a large twist sense bias. In contrast, in chloroform the oligomers adopt a random coil conformation. Surprisingly, the strong twist sense bias was determined to be highly time dependent and is partially attributed to intermolecular aggregation. PMID- 10841472 TI - Oxidative diphosphonylation of 1,4-dihydropyridines and pyridinium salts AB - An oxidative double phosphonylation of dihydropyridines 1 and pyridinium salts 2 is achieved through the use of dialkyl phosphites, DDQ, and triethylamine. Acceptable to good yields of 2, 6-diphosphonylated-1,2-dihydropyridines 3 are obtained in a one-pot reaction involving tandem nucleophilic addition/oxidation processes. Isomerization of 3 to the more stable 2,4-diphosphonylated-1, 4 dihydropyridine 4 was observed in some cases. PMID- 10841471 TI - Catalytic, asymmetric synthesis of the C(1)(')-C(10)(') segment of pamamycin 621A. AB - [reaction--see text] This paper describes a catalytic, asymmetric approach to the C(1)(')-C(10)(') segment of pamamycin 621A. We synthesize this segment in a convergent manner, with each of the coupling partners ultimately deriving from enantiomerically enriched methylketene dimer. PMID- 10841473 TI - Synthesis, structure proof, and biological activity of epothilone cyclopropanes. AB - [structure--see text] A semisynthetic route to epothilone cyclopropanes from epothilones A and B is described. Of significance, the deoxygenation of the 12, 13-epoxide to give the corresponding olefin was achieved with high efficiency. The title compounds (8, 9) were active in both tubulin polymerization and cytotoxicity assays, which is in direct contrast to a previously published report. These results provide further evidence that the role of the 12,13-epoxide of epothilones is largely conformational and argue against some of the current pharmacophore models. PMID- 10841474 TI - Solvolytic enolization of scytalone. AB - [reaction--see text] The major conformation of scytalone has an envelope shape with C3 forming the flap and the C3 hydroxyl in the equatorial position as determined by quantum mechanical calculations and corroborated by NMR. The C2 axial pro-R is slower to exchange with solvent than the equatorial pro-S hydrogen. Modeling the transition state for enolate formation points to a deprotonation through the flipped envelope conformation in which the C3-hydroxyl and the C2 pro-S hydrogen are axial. PMID- 10841475 TI - New phototriggers: extending the p-hydroxyphenacyl pi-pi absorption range. AB - [equation--see text] Introducing 3-methoxy or 3,5-dimethoxy substituents on the 4 hydroxyphenacyl (pHP) photoremovable protecting group has been explored with two excitatory gamma-amino acids, L-glutamic acid and gamma-amino butyric acid (GABA). These substituents significantly extend the absorption range of the pHP chromophore, e.g., the tail of absorption bands of 2a,b extend above 400 nm, well beyond the absorptions of aromatic amino acids and nucleotides. Irradiation releases the amino acids with rate constants of approximately 10(7) s(-)(1) and appearance efficiencies (Phi(app)) of 0.03-0.04. The photoproducts are formed through the pHP excited triplet and are primarily products of photoreduction and photohydrolysis. 1a,b also rearranged to the phenylacetic acid 3. PMID- 10841477 TI - Mechanistic studies of radical-based processes. Use and misuse of EPR spectroscopy AB - The identification of the species observed by EPR spectroscopy in the SmI(2) promoted coupling of N-(N', N'-dialkylaminoalkyl)benzotriazoles as substituted benzyl radicals is disputed on the basis of results obtained studying the model radical derived from tribenzylamine. PMID- 10841476 TI - Modular construction of dendritic carbosilanes. Organization Of dendrimer connectivity around bifunctional precursors that are adapted for sequential convergent and divergent propagative steps AB - Regiospecific hydrosilylation of 1-bromo-4-(prop-2-enyl)benzene offers an efficient route to molecular building block precursors that can accommodate sequential divergent and convergent steps for dendritic extension, establishing a modular methodology for assembly and organization of connectivity used for synthesis of modified carbosilane dendrimers including 14. PMID- 10841478 TI - Synthesis of 2-substituted-2,3- dihydro-1,4-dioxino[2,3-b]pyridine derivatives. AB - [reaction--see text] A variety of 2-substituted-2,3-dihydro-1,4-dioxino[2,3 b]pyridines B have been synthesized from the readily available 2-nitro-3 oxiranylmethoxypyridine 1 via a Smiles rearrangement. We demonstrate how variations of reaction conditions affect the product distribution of A and B. PMID- 10841480 TI - Supramolecular chirogenesis in bis(zinc porphyrin): An absolute configuration probe highly sensitive to guest structure AB - The role of the ligand's structure and absolute configuration in the supramolecular chirality induction in achiral bis(zinc porphyrin) has been studied. The amines with bulkier substituents resulted in stronger CD signals due to increased helical displacement in the anti conformer. All the amines with an R absolute configuration gave a negative first Cotton effect and positive second Cotton effect, while the ligands with an S absolute configuration produced CD signals with opposite signs due to formation of the left- and right-handed screw diastereomers, respectively. PMID- 10841479 TI - Synthesis and reactivity of conformationally locked alpha-aminoorganostannanes and alpha-aminoorganolithiums. Discovery of a surprising configurational requirement for transmetalation. AB - [equation--see text] 2-Tributylstannyl-N-methylpiperidines that are conformationally locked by a 4-tert-butyl substituent were evaluated in transmetalations (Sn-Li exchange) and reactions with electrophiles. When the tin is equatorial, transmetalation occurs smoothly as does reaction with carbonyl electrophiles. Alkyl halides seem to undergo single electron transfer reactions, affording nonselective alkylation products, along with products of radical disproportionation. In a surprise, an axially oriented tin failed to transmetalate, suggesting that a synclinal relationship between the nitrogen lone pair and the carbon-tin bond is a requirement for transmetalation. PMID- 10841481 TI - 2,5-Dimethylphenacyl as a new photoreleasable protecting group for carboxylic acids AB - The 2,5-dimethylphenacyl chromophore, a new photoremovable protecting group for carboxylic acids, is proposed. Direct photolysis of various 2,5-dimethylphenacyl esters in benzene or methanol at 254-366 nm leads to the formation of the corresponding carboxylic acids in almost quantitative isolated yields. The photodeprotection is based on efficient intramolecular hydrogen abstraction without the necessity of introducing a photosensitizer. PMID- 10841483 TI - Mannich-type reaction with trifluoromethylated N,O-hemiacetal: facile preparation of beta-amino-beta-trifluoromethyl carbonyl compounds AB - On treatment of silyl enolates and an N,O-hemiacetal, derived from trifluoroacetaldehyde ethyl hemiacetal and p-anisidine, with GaCl(3) (0.2 equiv) and C(6)H(5)COCl (0.2 equiv) in propionitrile, Mannich-type reaction took place smoothly to afford beta-amino-beta-trifluoromethyl carbonyl compounds in high yields. PMID- 10841482 TI - A convergent synthesis of (+)-cryptophycin B, a potent antitumor macrolide from Nostoc sp. cyanobacteria. AB - [structure--see text] An efficient and highly stereoselective synthesis of cryptophycin B (2), a potent cytotoxic agent, is described. The ester-derived titanium-enolate-mediated syn-aldol reaction was employed to generate the stereocenters C(5) and C(6). The route is convergent and provides a convenient access to the synthesis of structural variants of cryptophycin B as well as members of its family. PMID- 10841484 TI - Solid phase syntheses of polyamine toxins HO-416b and PhTX-433. Use of an efficient polyamide reduction strategy that facilitates access to branched analogues. AB - [structure--se text] Polyamine toxins HO-416b (1) and PhTX-433 (2) isolated from the venom of insects are important lead compounds in neuropharmacology. Their total synthesis has been achieved on a trityl derivatized resin in good yield and purity using a mild borane reduction protocol to access the polyamine chains from polyamide precursors. The synthesis of PhTX isomer 3 demonstrates the potential of this strategy for the generation of branched analogues. PMID- 10841485 TI - New chiral binaphthyl building blocks: synthesis of the first optically active tetrathiafulvalene dimers AB - New enantiomerically pure binaphthyl derivatives bearing triphenylphosphine or phosphonate groups have been synthesized and used as building blocks to prepare the first optically active tetrathiafulvalene dimers. PMID- 10841486 TI - Pyrrolidines from beta-aminoselenides via radical cyclization. Diastereoselectivity control by the N-substituent AB - N-Allyl-beta-aminoalkyl phenyl selenides-precursors of 3-aza-5-hexenyl radicals were prepared by ring opening of N-allylaziridines with benzeneselenol under acidic conditions or by sodium cyanoborohydride reduction of N-allylimines of alpha-phenylselenenyl ketones. The effect of various N-protective groups (acyl, sulfonyl, or phosphinoyl) on diastereoselectivity in thermally or photochemically initiated 3-aza-5-hexenyl reductive radical cyclization was studied. Whereas N unprotected derivatives afforded trans-2,4-disubstituded pyrrolidines with good selectivity, the diphenylphosphinoyl group directed cyclization to occur in a highly cis-selective manner. PMID- 10841487 TI - Remarkable O(2)-effect in 1,4-additions of diethylzinc to 6-acyloxy-2H-pyran 3(6H)-ones and 6-alkoxy-2H-pyran-3(6H)-ones AB - Under the influence of air, a facile 1,4-addition of diethylzinc to acyloxypyranones and alkoxypyranones 1 takes place. Reaction of diethylzinc with molecular oxygen provides EtOOZnEt, which catalyzes the addition of diethylzinc. PMID- 10841488 TI - First domino mukaiyama-aldol cyclizations of 1, 3-Bis(trimethylsiloxy)-1,3 butadienes with 1,2-diketones AB - The first domino Mukaiyama-aldol cyclization of 1, 3-bis(trimethylsiloxy)-1,3 dienes with enolizable 1,2-diketones provides a convenient and regioselective access to substituted cyclopent-2-en-1-ones. PMID- 10841489 TI - A new strategy for cyclopentenone synthesis. AB - [reaction--see text] A new strategy for the synthesis of 2,3-disubstituted cyclopentenones emerges from two key reactions-the ruthenium-catalyzed three component coupling of an equivalent of HBr, an alkyne, and a vinyl ketone and the Ni-Cr Barbier type reaction. As a result, these important structures are readily accessed from an alkyne and a vinyl ketone (which derive directly from carboxylic acids). Syntheses of tetrahydrodicranenone B and rosaprostol illustrate the new strategy. PMID- 10841490 TI - Cyclodidemniserinol trisulfate, a sulfated serinolipid from the Palauan ascidian Didemnum guttatum that inhibits HIV-1 integrase. AB - [structure--see text] Bioassay-guided fractionation of extracts of the Palauan ascidian Didemnum guttatum led to the isolation of cyclodidemniserinol trisulfate (1) as an inhibitor of HIV-1 integrase, which is an attractive target for anti retroviral chemotherapy. The structure of cyclodidemniserinol trisulfate (1), the stereochemistry of which was only partially determined, was elucidated by interpretation of NMR and mass spectral data. PMID- 10841491 TI - A new and practical five-carbon component for metal-catalyzed AB - Described herein is an efficient preparative scale synthesis of 1-(2 methyoxyethoxy)-1-vinylcyclopropane and the investigation of the utility of this reagent as a new five-carbon component in metal-catalyzed [5 + 2] cycloadditions. A new cycloaddition procedure is also described that proceeds up to 12-fold faster and with 10-fold less catalyst than previously described, providing cycloheptenones in many cases in minutes and in isolated yields of 75-97%. The procedure is readily conducted on a small or large scale (up to 100 mmol thus far). PMID- 10841493 TI - Vanadium-catalyzed asymmetric cyanohydrin synthesis AB - Based on a mechanistic understanding of asymmetric cyanohydrin synthesis catalyzed by chiral titanium-salen complexes, a new catalyst based on vanadium(IV) has been developed. The chiral (salen)VO catalyst is more enantioselective than the titanium-based systems, 0.1 mol % of the catalyst being sufficient to convert aromatic and aliphatic aldehydes into the corresponding trimethylsilyl ethers of cyanohydrins with 68-95% enantiomeric excess at room temperature. PMID- 10841492 TI - A novel route to preussomerins via 2-arylacetal anions. AB - [reaction--see text] Dimerization of salicylaldehydes provided 6H,12H-6,12 epoxydibenzo[b, f][1,5]dioxocins in multigram quantities. Deprotonation allylation of the benzylic acetals followed by further functionalization of the diallyl derivative and double Friedel-Crafts cyclization gave a novel preussomerin analogue which possessed the full carbon skeleton of the natural products. PMID- 10841494 TI - Synthesis of periphery-functionalized dendritic molecules using polylithiated dendrimers as starting material AB - A general method for the functionalization of Si-Cl terminated carbosilane dendritic molecules via organolithium or organomagnesium reagents is described. Quantitative exchange of the bromine atoms of 4-bromophenyl-functionalized dendrimers affords polylithiated species that are valuable starting materials for further functionalization, e.g., into pyridyl alcohols. The latter were successfully applied as catalyst precursors in a ruthenium-mediated ring-closure metathesis reaction. PMID- 10841495 TI - Regiocontrol in an intramolecular Schmidt reaction: total synthesis of (+) aspidospermidine. AB - [reaction--see text] A total synthesis of (+)-aspidospermidine (1) is described, featuring an intramolecular Schmidt reaction as the key step. The effects of stereochemistry and protecting group status on the regio- and chemoselectivity of this reaction were examined. PMID- 10841496 TI - Kinetic control in the palladium-catalyzed synthesis of C(2)-symmetric hexabenzotriphenylene. A conformational study AB - The hitherto unisolated, thermodynamically unstable C(2)-symmetric conformer of hexabenzotriphenylene (1) has been efficiently synthesized by palladium-catalyzed cyclotrimerization of 9, 10-didehydrophenanthrene (4). The barriers to conformational interconversion of 1 are examined experimentally and by computational studies. PMID- 10841497 TI - En route to a plant scale synthesis of the promising antitumor agent 12,13 desoxyepothilone B. AB - [reaction--see text] Efficient and processable syntheses of key building blocks of the antitumor agent 12,13-desoxyepothilone B (dEpoB) by catalytic asymmetric induction are herein described. PMID- 10841498 TI - On the total synthesis and preliminary biological evaluations of 15(R) and 15(S) aza-dEpoB: a Mitsunobu inversion at C15 in pre-epothilone fragments. AB - [reaction-see text] The syntheses of two epothilone analogues, 15(S)-aza-12,13 desoxyepothilone B and the epimeric 15(R)-aza-12,13-desoxyepothilone B, are described. A Mitsunobu inversion was utilized for elaboration of pre-epothilone fragments to the corresponding macrolactam. Tubulin binding and cytotoxicity profiles of these analogues are presented. PMID- 10841499 TI - A Rhodium(II) catalytic approach to the synthesis of ethers of a minor component in a tautomeric set. AB - [equation--see text] The Rh(II)-catalyzed reaction of diazoacetic esters with various carbonyl compounds is an effective method for the synthesis of acetic ester ethers of the corresponding enol forms. PMID- 10841500 TI - A facile synthetic route to a third-generation dendrimer with generation-specific functional aryl bromides AB - The synthesis of three third-generation dendrimers that selectively carry one aryl bromide functional group in the first, second, or third generation, respectively, is described. These functions, regardless of their location, can be chemically modified by Suzuki cross-coupling chemistry with p-tert-butylbenzene boronic ester. PMID- 10841501 TI - Stem cells near the century mark. PMID- 10841502 TI - Gut instincts: thoughts on intestinal epithelial stem cells. PMID- 10841503 TI - PARP alleles and SLE: failure to confirm association with disease susceptibility. PMID- 10841504 TI - Searching for genes in complex diseases: lessons from systemic lupus erythematosus. PMID- 10841505 TI - Fibronectin peptides in cell migration and wound repair. PMID- 10841506 TI - Is cyclooxygenase-2 the alpha and the omega in cancer? PMID- 10841507 TI - Viral antichemokines: from pathogenesis to drug discovery. PMID- 10841508 TI - Proteinases in cardiovascular aneurysms and rupture: targets for therapy? PMID- 10841509 TI - A role for the serotonin transporter in hypoxia-induced pulmonary hypertension. PMID- 10841510 TI - Angiotensin II, atherosclerosis, and aortic aneurysms. PMID- 10841511 TI - Transplanted cord blood-derived endothelial precursor cells augment postnatal neovascularization. AB - Endothelial precursor cells (EPCs) have been identified in adult peripheral blood. We examined whether EPCs could be isolated from umbilical cord blood, a rich source for hematopoietic progenitors, and whether in vivo transplantation of EPCs could modulate postnatal neovascularization. Numerous cell clusters, spindle shaped and attaching (AT) cells, and cord-like structures developed from culture of cord blood mononuclear cells (MNCs). Fluorescence-trace experiments revealed that cell clusters, AT cells, and cord-like structures predominantly were derived from CD34-positive MNCs (MNC(CD34+)). AT cells and cell clusters could be generated more efficiently from cord blood MNCs than from adult peripheral blood MNCs. AT cells incorporated acetylated-LDL, released nitric oxide, and expressed KDR, VE-cadherin, CD31, and von Willebrand factor but not CD45. Locally transplanted AT cells survived and participated in capillary networks in the ischemic tissues of immunodeficient nude rats in vivo. AT cells thus had multiple endothelial phenotypes and were defined as a major population of EPCs. Furthermore, laser Doppler and immunohistochemical analyses revealed that EPC transplantation quantitatively augmented neovascularization and blood flow in the ischemic hindlimb. In conclusion, umbilical cord blood is a valuable source of EPCs, and transplantation of cord blood-derived EPCs represents a promising strategy for modulating postnatal neovascularization. PMID- 10841513 TI - Mouse embryogenesis requires the tissue factor extracellular domain but not the cytoplasmic domain. AB - Recent studies indicate that tissue factor (TF) acts in embryogenesis, metastasis, and angiogenesis. Three independent groups showed that targeted disruption of the murine TF (mTF) gene results in 90% lethality of mTF null embryos at embryonic days 9. 5-10.5. We have demonstrated that expression of wild type human TF (hTF) from a minigene rescues the embryonic lethality of mTF null embryos. To investigate the role of TF in embryogenesis, we made mutant hTF minigenes whose products either bound FVII/VIIa at a reduced level or lacked the cytoplasmic domain. Two independent transgenic lines expressing the hTF extracellular domain mutant failed to rescue the embryonic lethality of mTF null embryos, suggesting that FVII/VIIa binding by TF, proteolytic activity by the TF/FVIIa complex, or both were required for embryogenesis. In contrast, two transgenic lines expressing the hTF cytoplasmic domain mutant rescued the embryonic lethality of mTF null embryos, indicating that the cytoplasmic domain of TF was not required for embryogenesis. We propose that TF/FVIIa-dependent extracellular protease activity is required for embryogenesis. PMID- 10841512 TI - The PHSRN sequence induces extracellular matrix invasion and accelerates wound healing in obese diabetic mice. AB - The PHSRN sequence of the plasma fibronectin (pFn) cell-binding domain induces human keratinocytes and fibroblasts to invade the naturally serum-free extracellular matrices of sea urchin embryos. The potency of acetylated, amidated PHSRN (Ac-PHSRN-NH(2)) is significantly increased, making it more active on a molar basis than the 120-kDa cell-binding domain of pFn. Arginine is important to this activity because PHSAN and PHSEN are inactive, as is a randomized sequence peptide, Ac-HSPNR-NH(2). One treatment with Ac-PHSRN-NH(2) stimulates reepithelialization and contraction of dermal wounds in healing-impaired, obese diabetic C57BL6/KsJ db/db mice. Wound closure is equally rapid in treated db/db and db/+ mice and may be more rapid than in untreated nondiabetic db/+ littermates. In contrast, treatment with either Ac-HSPNR-NH(2) or normal saline (NS) has no effect. Analysis of sectioned db/db wounds shows that, in contrast to treatment with Ac-HSPNR-NH(2) or NS, a single Ac-PHSRN-NH(2) treatment stimulates keratinocyte and fibroblast migration into wounds, enhances fibroplasia and vascularization in the provisional matrix, and stimulates the formation of prominent fibers that may be associated with wound contraction. PMID- 10841514 TI - Attenuated hypoxic pulmonary hypertension in mice lacking the 5-hydroxytryptamine transporter gene. AB - Hypoxia is a well-recognized stimulus for pulmonary blood vessel remodeling and pulmonary hypertension development. One mechanism that may account for these effects is the direct action of hypoxia on the expression of specific genes involved in vascular smooth muscle cell (SMC) proliferation. Previous studies demonstrated that the serotonin (5-hydroxytryptamine; 5-HT) transporter (5-HTT) mediates the mitogenic activity of 5-HT in pulmonary vascular SMCs and is overexpressed during hypoxia. Thus, 5-HT-related mitogenic activity is increased during hypoxia. Here, we report that mice deficient for 5-HTT (5-HTT(-/-)) developed less hypoxic pulmonary hypertension and vascular remodeling than paired 5-HTT(+/+) controls. When maintained under normoxia, 5-HTT(-/-)-mutant mice had normal hemodynamic parameters, low blood 5-HT levels, deficient platelet 5-HT uptake, and unchanged blood levels of 5-hydroxyindoleacetic acid, a metabolite of 5-HT. After exposure to 10% O(2) for 2 or 5 weeks, the number and medial wall thickness of muscular pulmonary vessels were reduced in hypoxic 5-HTT(-/-) mice as compared with wild-type paired controls. Concomitantly, right ventricular systolic pressure was lower and right ventricle hypertrophy less marked in the mutant mice. This occurred despite potentiation of acute hypoxic pulmonary vasoconstriction in the 5-HTT(-/-) mice. These data further support a key role of 5-HTT in hypoxia-induced pulmonary vascular SMC proliferation and pulmonary hypertension. PMID- 10841515 TI - Reduction of globotriaosylceramide in Fabry disease mice by substrate deprivation. AB - We used a potent inhibitor of glucosylceramide synthase to test whether substrate deprivation could lower globotriaosylceramide levels in alpha-galactosidase A (alpha-gal A) knockout mice, a model of Fabry disease. C57BL/6 mice treated twice daily for 3 days with D-threo-1-ethylendioxyphenyl-2-palmitoylamino-3-pyrrolidi no-propanol (D-t-EtDO-P4) showed a concentration-dependent decrement in glucosylceramide levels in kidney, liver, and spleen. A single intraperitoneal injection of D-t-EtDO-P4 resulted in a 55% reduction in renal glucosylceramide, consistent with rapid renal glucosylceramide metabolism. A concentration dependent decrement in renal and hepatic globotriaosylceramide levels was observed in alpha-Gal A(-) males treated for 4 weeks with D-t-EtDO-P4. When 8 week-old alpha-Gal A(-) males were treated for 8 weeks with 10 mg/kg twice daily, renal globotriaosylceramide fell to below starting levels, consistent with an alpha-galactosidase A-independent salvage pathway for globotriaosylceramide degradation. Complications observed with another glucosylceramide synthase inhibitor, N-butyldeoxynojirimycin, including weight loss and acellularity of lymphatic organs, were not observed with D-t-EtDO-P4. These data suggest that Fabry disease may be amenable to substrate deprivation therapy. PMID- 10841516 TI - Endosomal processing limits gene transfer to polarized airway epithelia by adeno associated virus. AB - The restriction of viral receptors and coreceptors to the basolateral surface of airway epithelial cells has been blamed for the inefficient transfer of viral vectors to the apical surface of this tissue. We now report, however, that differentiated human airway epithelia internalize rAAV type-2 virus efficiently from their apical surfaces, despite the absence of known adeno-associated virus-2 (AAV-2) receptors or coreceptors at these sites. The dramatically lower transduction efficiency of rAAV infection from the apical surface of airway cells appears to result instead from differences in endosomal processing and nuclear trafficking of apically or basolaterally internalized virions. AAV capsid proteins are ubiquitinated after endocytosis, and gene transfer can be significantly enhanced by proteasome or ubiquitin ligase inhibitors. Tripeptide proteasome inhibitors increased persistent rAAV gene delivery from the apical surface >200-fold, to a level nearly equivalent to that achieved with basolateral infection. In vivo application of proteasome inhibitor in mouse lung augmented rAAV gene transfer from undetectable levels to a mean of 10.4 +/- 1.6% of the epithelial cells in large bronchioles. Proteasome inhibitors also increased rAAV 2-mediated gene transfer to the liver tenfold, but they did not affect transduction of skeletal or cardiac muscle. These findings suggest that tissue specific ubiquitination of viral capsid proteins interferes with rAAV-2 transduction and provides new approaches to circumvent this barrier for gene therapy of diseases such as cystic fibrosis. PMID- 10841517 TI - Host cyclooxygenase-2 modulates carcinoma growth. AB - Cyclooxygenase-2 (COX-2; Ptgs2) acts as a tumor promoter in rodent models for colorectal cancer, but its precise role in carcinogenesis remains unclear. We evaluated the contribution of host-derived COX-1 and COX-2 in tumor growth using both genetic and pharmacological approaches. Lewis lung carcinoma (LLC) cells grow rapidly as solid tumors when implanted in C57BL/6 mice. We found that tumor growth was markedly attenuated in COX-2(-/-), but not COX-1(-/-) or wild-type mice. Treatment of wild-type C57BL/6 mice bearing LLC tumors with a selective COX 2 inhibitor also reduced tumor growth. A decrease in vascular density was observed in tumors grown in COX-2(-/-) mice when compared with those in wild-type mice. Because COX-2 is expressed in stromal fibroblasts of human and rodent colorectal carcinomas, we evaluated COX-2(-/-) mouse fibroblasts and found a 94% reduction in their ability to produce the proangiogenic factor, VEGF. Additionally, treatment of wild-type mouse fibroblasts with a selective COX-2 inhibitor reduced VEGF production by 92%. PMID- 10841518 TI - Antagonizing the parathyroid calcium receptor stimulates parathyroid hormone secretion and bone formation in osteopenic rats. AB - Parathyroid hormone (PTH) is an effective bone anabolic agent, but it must be administered parenterally. An orally active anabolic agent would provide a valuable alternative for treating osteoporosis. NPS 2143 is a novel, selective antagonist (a "calcilytic") of the parathyroid cell Ca(2+) receptor. Daily oral administration of NPS 2143 to osteopenic ovariectomized (OVX) rats caused a sustained increase in plasma PTH levels, provoking a dramatic increase in bone turnover but no net change in bone mineral density. Concurrent oral administration of NPS 2143 and subcutaneous infusion of 17beta-estradiol also resulted in increased bone turnover. However, the antiresorptive action of estrogen decreased the extent of bone resorption stimulated by the elevated PTH levels, leading to an increase in bone mass compared with OVX controls or to either treatment alone. Despite the sustained stimulation to the parathyroid gland, parathyroid cells did not undergo hyperplasia. These data demonstrate that an increase in endogenous PTH secretion, induced by antagonism of the parathyroid cell Ca(2+) receptor with a small molecule, leads to a dramatic increase in bone turnover, and they suggest a novel approach to the treatment of osteoporosis. PMID- 10841519 TI - Angiotensin II promotes atherosclerotic lesions and aneurysms in apolipoprotein E deficient mice. AB - Increased plasma concentrations of angiotension II (Ang II) have been implicated in atherogenesis. To examine this relationship directly, we infused Ang II or vehicle for 1 month via osmotic minipumps into mature apoE(-/-) mice. These doses of Ang II did not alter arterial blood pressure, body weight, serum cholesterol concentrations, or distribution of lipoprotein cholesterol. However, Ang II infusions promoted an increased severity of aortic atherosclerotic lesions. These Ang II-induced lesions were predominantly lipid-laden macrophages and lymphocytes; moreover, Ang II promoted a marked increase in the number of macrophages present in the adventitial tissue underlying lesions. Unexpectedly, pronounced abdominal aortic aneurysms were present in apoE(-/-) mice infused with Ang II. Sequential sectioning of aneurysmal abdominal aorta revealed two major characteristics: an intact artery that is surrounded by a large remodeled adventitia, and a medial break with pronounced dilation and more modestly remodeled adventitial tissue. Although no atherosclerotic lesions were visible at the medial break point, the presence of hyperlipidemia was required because infusions of Ang II into apoE(+/+) mice failed to generate aneurysms. These results demonstrate that increased plasma concentrations of Ang II have profound and rapid effects on vascular pathology when combined with hyperlipidemia, in the absence of hemodynamic influences. PMID- 10841520 TI - The viral anti-inflammatory chemokine-binding protein M-T7 reduces intimal hyperplasia after vascular injury. AB - Chemokines and IFN-gamma function as central regulators of inflammatory responses to vascular injury. Both classes of cytokines are upregulated during restenosis, a response to vascular injury that leads to recurrent atherosclerotic plaque growth, but the relative impact of each class of cytokines remains undetermined. M-T7 is a secreted myxoma viral immunomodulatory glycoprotein that functions both as a species-specific inhibitor of rabbit IFN-gamma and as a chemokine-binding protein, interacting with a wide range of C, C-C, and C-X-C chemokines in a species-nonspecific fashion. We wished to (a) assess the efficacy of purified M T7 protein in inhibiting intimal hyperplasia after angioplasty injury and (b) exploit unique species-specific functions of M-T7 in order to judge the relative importance of each cytokine class on plaque growth. Anesthetized New Zealand white rabbits and Sprague-Dawley rats received either M-T7 or control at the time of arterial angioplasty injury. Histological analysis at 28 days demonstrated significant reductions in intimal hyperplasia with M-T7 treatment in both models, with an associated early inhibition of inflammatory cell invasion. Purified M-T7 protein inhibits intimal hyperplasia after angioplasty injury in a species nonspecific fashion, thus implicating the chemokine-binding activity as more critical for prevention of plaque growth after vascular injury. PMID- 10841521 TI - Melanoma immunotherapy by targeted IL-2 depends on CD4(+) T-cell help mediated by CD40/CD40L interaction. AB - The induction of tumor-protective immunity against malignancies remains a major challenge in cancer immunotherapy. A novel, humanized anti-ganglioside-GD(2)-IL-2 immunocytokine (hu14.18-IL-2) induced CD8(+) T cells to eradicate established pulmonary metastases of B78-D14 murine melanoma, in a process that required help by CD4(+) T cells and was mediated by the CD40/CD40 ligand (CD40L) interaction. The anti-tumor effect was diminished in mice deficient in CD4(+) T-cells. Three lines of evidence show that CD4(+) T-cell help was mediated by CD40/CD40L interaction but not by endogenous IL-2 production. First, the hu14.18-IL-2 induced anti-tumor response is partially abrogated in C57BL/6J CD40L knockout (KO) mice in contrast to C57BL/6J IL-2 KO animals, in which the immunocytokine was completely effective. Second, partial abrogation of the anti-tumor effect is induced with anti-CD40L antibodies to the same extent as with CD4(+) T-cell depletion. Third, a complete anti-tumor response induced by hu14.18-IL-2 can be reconstituted in C57BL/6J CD40L KO mice by simultaneous stimulation with an anti CD40 mAb. These results suggest that help provided by CD4(+) T cells via CD40/CD40L interactions in our tumor model is crucial for effective immunotherapy with an IL-2 immunocytokine. PMID- 10841522 TI - Regulation of the vascular extracellular superoxide dismutase by nitric oxide and exercise training. AB - The bioactivity of endothelium-derived nitric oxide (NO) reflects its rates of production and of inactivation by superoxide (O(2)(*-)), a reactive species dismutated by extracellular superoxide dismutase (ecSOD). We have now examined the complementary hypothesis, namely that NO modulates ecSOD expression. The NO donor DETA-NO increased ecSOD expression in a time- and dose-dependent manner in human aortic smooth muscle cells. This effect was prevented by the guanylate cyclase inhibitor ODQ and by the protein kinase G (PKG) inhibitor Rp-8-CPT-cGMP. Expression of ecSOD was also increased by 8-bromo-cGMP, but not by 8-bromo-cAMP. Interestingly, the effect of NO on ecSOD expression was prevented by inhibition of the MAP kinase p38 but not of the MAP kinase kinase p42/44, suggesting that NO modulates ecSOD expression via cGMP/PKG and p38MAP kinase-dependent pathways, but not through p42/44MAP kinase. In aortas from mice lacking the endothelial nitric oxide synthase (eNOS), ecSOD was reduced more than twofold compared to controls. Treadmill exercise training increased eNOS and ecSOD expression in wild-type mice but had no effect on ecSOD expression in mice lacking eNOS, suggesting that this effect of exercise is meditated by endothelium-derived NO. Upregulation of ecSOD expression by NO may represent an important feed-forward mechanism whereby endothelial NO stimulates ecSOD expression in adjacent smooth muscle cells, thus preventing O(2)(*-)-mediated degradation of NO as it traverses between the two cell types. PMID- 10841523 TI - Targeted gene disruption of matrix metalloproteinase-9 (gelatinase B) suppresses development of experimental abdominal aortic aneurysms. AB - Abdominal aortic aneurysms represent a life-threatening condition characterized by chronic inflammation, destructive remodeling of the extracellular matrix, and increased local expression of matrix metalloproteinases (MMPs). Both 92-kD gelatinase (MMP-9) and macrophage elastase (MMP-12) have been implicated in this disease, but it is not known if either is necessary in aneurysmal degeneration. We show here that transient elastase perfusion of the mouse aorta results in delayed aneurysm development that is temporally associated with transmural mononuclear inflammation, increased local production of several elastolytic MMPs, and progressive destruction of the elastic lamellae. Elastase-induced aneurysmal degeneration was suppressed by treatment with a nonselective MMP inhibitor (doxycycline) and by targeted gene disruption of MMP-9, but not by isolated deficiency of MMP-12. Bone marrow transplantation from wild-type mice prevented the aneurysm-resistant phenotype in MMP-9-deficient animals, and wild-type mice acquired aneurysm resistance after transplantation from MMP-9-deficient donors. These results demonstrate that inflammatory cell expression of MMP-9 plays a critical role in an experimental model of aortic aneurysm disease, suggesting that therapeutic strategies targeting MMP-9 may limit the growth of small abdominal aortic aneurysms. PMID- 10841524 TI - Na(+)-dependent transporters mediate HCO(3)(-) salvage across the luminal membrane of the main pancreatic duct. AB - To study the roles of Na(+)-dependent H(+) transporters, we characterized H(+) efflux mechanisms in the pancreatic duct in wild-type, NHE2(-/-), and NHE3(-/-) mice. The pancreatic duct expresses NHE1 in the basolateral membrane, and NHE2 and NHE3 in the luminal membrane, but does not contain NHE4 or NHE5. Basolateral Na(+)-dependent H(+) efflux in the microperfused duct was inhibited by 1.5 microM of the amiloride analogue HOE 694, consistent with expression of NHE1, whereas the luminal activity required 50 microM HOE 694 for effective inhibition, suggesting that the efflux might be mediated by NHE2. However, disruption of NHE2 had no effect on luminal transport, while disruption of the NHE3 gene reduced luminal Na(+)-dependent H(+) efflux by approximately 45%. Notably, the remaining luminal Na(+)-dependent H(+) efflux in ducts from NHE3(-/-) mice was inhibited by 50 microM HOE 694. Hence, approximately 55% of luminal H(+) efflux (or HCO(3)(-) influx) in the pancreatic duct is mediated by a novel, HOE 694-sensitive, Na(+) dependent mechanism. H(+) transport by NHE3 and the novel transporter is inhibited by cAMP, albeit to different extents. We propose that multiple Na(+) dependent mechanisms in the luminal membrane of the pancreatic duct absorb Na(+) and HCO(3)(-) to produce a pancreatic juice that is poor in HCO(3)(-) and rich in Cl(-) during basal secretion. Inhibition of the transporters during stimulated secretion aids in producing the HCO(3)(-)-rich pancreatic juice. PMID- 10841525 TI - The persistence of alien genomes. PMID- 10841526 TI - Glioblastoma multiforme: the terminator. PMID- 10841528 TI - LINE-1 elements and X chromosome inactivation: a function for "junk" DNA? PMID- 10841527 TI - A multiplicity of muscarinic mechanisms: enough signaling pathways to take your breath away. PMID- 10841529 TI - Retrotransposon-mediated genome evolution on a local ecological scale. PMID- 10841530 TI - Optimal prediction of stiff oscillatory mechanics. AB - We consider many-body problems in classical mechanics where a wide range of time scales limits what can be computed. We apply the method of optimal prediction to obtain equations that are easier to solve numerically. We demonstrate by examples that optimal prediction can reduce the amount of computation needed to obtain a solution by several orders of magnitude. PMID- 10841531 TI - Temperature profiles and bathymetry of some high mountain lakes. AB - The ice cover in high mountain lakes breaks up and disappears in about an hour, in part because it has been divided into fragile vertical spindles, which are individual crystals. Contributing to this process are vertical holes in the ice remaining after particles of dust melt downward as they are warmed by the sun. Temperature profiles of lakes in the Wind River Mountains of Wyoming include those of five lakes in the high wilderness. All are isothermal at or near 5 degrees C below a depth of 20 to 30 m, regardless of elevation or lake depth. Large lakes have a deeper mixed layer than do small ones because of longer fetch and thus more effective wind shear. PMID- 10841532 TI - Synthetic receptors as models for alkali metal cation-pi binding sites in proteins. AB - The alkali metal cations Na(+) and K(+) have several important physiological roles, including modulating enzyme activity. Recent work has suggested that alkali metal cations may be coordinated by pi systems, such as the aromatic amino acid side chains. The ability of K(+) to interact with an aromatic ring has been assessed by preparing a family of synthetic receptors that incorporate the aromatic side chains of phenylalanine, tyrosine, and tryptophan. These receptors are constructed around a diaza-18-crown-6 scaffold, which serves as the primary binding site for an alkali metal cation. The ability of the aromatic rings to coordinate a cation was determined by crystallizing each of the receptors in the presence of K(+) and by solving the solid state structures. In all cases, complexation of K(+) by the pi system was observed. When possible, the structures of the unbound receptors also were determined for comparison. Further proof that the aromatic ring makes an energetically favorable interaction with the cation was obtained by preparing a receptor in which the arene was perfluorinated. Fluorination of the arene reverses the electrostatics, but the aromaticity is maintained. The fluorinated arene rings do not coordinate the cation in the solid state structure of the K(+) complex. Thus, the results of the predicted electrostatic reversal were confirmed. Finally, the biological implications of the alkali metal cation-pi interaction are addressed. PMID- 10841533 TI - Atomic force microscopy captures quantized plastic deformation in gold nanowires. AB - Scanning probe microscopy has become a powerful tool to detect structural changes in small clusters of atoms. Herein, we use an atomic force microscope to measure the length of gold nanowire structures during extension and compression cycles. We have found that nanowires elongate under force in quantized steps of up to three integer multiples of 1.76 A and that they shorten spontaneously in steps of 1.52 A. Our results can be explained by the sliding of crystal planes within the gold nanowires creating stacking faults that change the local structure from face centered cubic to hexagonal close packed. Our data also show that there can be up to three simultaneous slip events, in good agreement with the tetrahedral arrangement of slip planes in a gold crystal. These experiments provide direct evidence for the mechanism underlying the plastic deformation of a nanowire. A similar approach can be used to examine the atomic events underlying the plastic failure of other metals and their alloys. PMID- 10841534 TI - Enhanced metabolism of halogenated hydrocarbons in transgenic plants containing mammalian cytochrome P450 2E1. AB - Chlorinated solvents, especially trichloroethylene (TCE), are the most widespread groundwater contaminants in the United States. Existing methods of pumping and treating are expensive and laborious. Phytoremediation, the use of plants for remediation of soil and groundwater pollution, is less expensive and has low maintenance; however, it requires large land areas and there are a limited number of suitable plants that are known to combine adaptation to a particular environment with efficient metabolism of the contaminant. In this work, we have engineered plants with a profound increase in metabolism of the most common contaminant, TCE, by introducing the mammalian cytochrome P450 2E1. This enzyme oxidizes a wide range of important pollutants, including TCE, ethylene dibromide, carbon tetrachloride, chloroform, and vinyl chloride. The transgenic plants had a dramatic enhancement in metabolism of TCE of up to 640-fold as compared with null vector control plants. The transgenic plants also showed an increased uptake and debromination of ethylene dibromide. Therefore, transgenic plants with this enzyme could be used for more efficient remediation of many sites contaminated with halogenated hydrocarbons. PMID- 10841535 TI - Rational design of nascent metalloenzymes. AB - Understanding the early genesis of new enzymatic functions is one of the challenges in protein design, mechanistic enzymology, and molecular evolution. We have experimentally mimicked starting points in this process by introducing primitive iron and oxygen binding sites at various locations in thioredoxin, a small protein lacking metal centers, by using computational design. These rudimentary active sites show emerging enzymatic activities that select to varying degrees between different oxygen chemistries. Even within these nascent enzymes, mechanisms by which different reactions are controlled can be discerned. These involve both stabilizing and destabilizing interactions imposed on the metal center by the surrounding protein matrix. PMID- 10841536 TI - Retrostructural analysis of metalloproteins: application to the design of a minimal model for diiron proteins. AB - De novo protein design provides an attractive approach for the construction of models to probe the features required for function of complex metalloproteins. The metal-binding sites of many metalloproteins lie between multiple elements of secondary structure, inviting a retrostructural approach to constructing minimal models of their active sites. The backbone geometries comprising the metal binding sites of zinc fingers, diiron proteins, and rubredoxins may be described to within approximately 1 A rms deviation by using a simple geometric model with only six adjustable parameters. These geometric models provide excellent starting points for the design of metalloproteins, as illustrated in the construction of Due Ferro 1 (DF1), a minimal model for the Glu-Xxx-Xxx-His class of dinuclear metalloproteins. This protein was synthesized and structurally characterized as the di-Zn(II) complex by x-ray crystallography, by using data that extend to 2.5 A. This four-helix bundle protein is comprised of two noncovalently associated helix-loop-helix motifs. The dinuclear center is formed by two bridging Glu and two chelating Glu side chains, as well as two monodentate His ligands. The primary ligands are mostly buried in the protein interior, and their geometries are stabilized by a network of hydrogen bonds to second-shell ligands. In particular, a Tyr residue forms a hydrogen bond to a chelating Glu ligand, similar to a motif found in the diiron-containing R2 subunit of Escherichia coli ribonucleotide reductase and the ferritins. DF1 also binds cobalt and iron ions and should provide an attractive model for a variety of diiron proteins that use oxygen for processes including iron storage, radical formation, and hydrocarbon oxidation. PMID- 10841537 TI - Crystal structure of RPB5, a universal eukaryotic RNA polymerase subunit and transcription factor interaction target. AB - Eukaryotic nuclei contain three different types of RNA polymerases (RNAPs), each consisting of 12-18 different subunits. The evolutionarily highly conserved RNAP subunit RPB5 is shared by all three enzymes and therefore represents a key structural/functional component of all eukaryotic RNAPs. Here we present the crystal structure of the RPB5 subunit from Saccharomyces cerevisiae. The bipartite structure includes a eukaryote-specific N-terminal domain and a C terminal domain resembling the archaeal RNAP subunit H. RPB5 has been implicated in direct protein-protein contacts with transcription factor IIB, one of the components of the RNAP(II) basal transcriptional machinery, and gene-specific activator proteins, such as the hepatitis B virus transactivator protein X. The experimentally mapped regions of RPB5 involved in these interactions correspond to distinct and surface-exposed alpha-helical structures. PMID- 10841538 TI - Solution structure of the RNA polymerase subunit RPB5 from Methanobacterium thermoautotrophicum. AB - RPB5 is an essential subunit of eukaryotic and archaeal RNA polymerases. It is a proposed target for transcription activator proteins in eukaryotes, but the mechanism of interaction is not known. We have determined the solution structure of the RPB5 subunit from the thermophilic archeon, Methanobacterium thermoautotrophicum. MtRBP5 contains a four-stranded beta-sheet platform supporting two alpha-helices, one on each side of the beta-sheet, resulting in an overall mushroom shape that does not appear to have any structural homologues in the structural database. The position and conservation of charged surface residues suggests possible modes of interaction with other proteins, as well as a rationale for the thermal stability of this protein. PMID- 10841539 TI - Zinc-bundle structure of the essential RNA polymerase subunit RPB10 from Methanobacterium thermoautotrophicum. AB - The RNA polymerase subunit RPB10 displays a high level of conservation across archaea and eukarya and is required for cell viability in yeast. Structure determination of this RNA polymerase subunit from Methanobacterium thermoautotrophicum reveals a topology, which we term a zinc-bundle, consisting of three alpha-helices stabilized by a zinc ion. The metal ion is bound within an atypical CX(2)CX(n)CC sequence motif and serves to bridge an N-terminal loop with helix 3. This represents an example of two adjacent zinc-binding Cys residues within an alpha-helix conformation. Conserved surface features of RPB10 include discrete regions of neutral, acidic, and basic residues, the latter being located around the zinc-binding site. One or more of these regions may contribute to the role of this subunit as a scaffold protein within the polymerase holoenzyme. PMID- 10841540 TI - Enantiomer discrimination illustrated by high-resolution crystal structures of the human nuclear receptor hRARgamma. AB - The human retinoic acid receptor (hRAR) is a member of the nuclear receptor superfamily that regulates the transcription of target genes in a ligand dependent manner. The three hRAR isotypes are targets for retinoids that are used in the treatment of various diseases, including breast cancer and skin diseases. Drug efficiency and safety depend on the pharmacological activity of enantiomers, which can differ because of the chiral environment generated by the target. We report the crystal structures of the hRARgamma ligand-binding domain bound to two enantiomers, the active BMS270394 and the inactive BMS270395, solved at 1.6 A and 1.7 A resolution, respectively. The crystal structures reveal that in both enantiomers, the hydroxyl moiety attached to the chiral center forms a hydrogen bond to the Met-272 sulfur atom, thus imposing a conformation of BMS270395 that differs significantly from that observed for BMS270394 and other known retinoids. BMS270395 adopts an energetically unfavorable conformation, accounting for its inactivity; in contrast, the conformation of BMS270394 is close to an energy minimum. Our high-resolution data allow rationalization of enantiomer discrimination by the receptor and provide a model system for the pharmacological properties of enantiomeric pairs. PMID- 10841541 TI - Functional implications from crystal structures of the conserved Bacillus subtilis protein Maf with and without dUTP. AB - Three-dimensional structures of functionally uncharacterized proteins may furnish insight into their functions. The potential benefits of three-dimensional structural information regarding such proteins are particularly obvious when the corresponding genes are conserved during evolution, implying an important function, and no functional classification can be inferred from their sequences. The Bacillus subtilis Maf protein is representative of a family of proteins that has homologs in many of the completely sequenced genomes from archaea, prokaryotes, and eukaryotes, but whose function is unknown. As an aid in exploring function, we determined the crystal structure of this protein at a resolution of 1.85 A. The structure, in combination with multiple sequence alignment, reveals a putative active site. Phosphate ions present at this site and structural similarities between a portion of Maf and the anticodon-binding domains of several tRNA synthetases suggest that Maf may be a nucleic acid binding protein. The crystal structure of a Maf-nucleoside triphosphate complex provides support for this hypothesis and hints at di- or oligonucleotides with either 5'- or 3'-terminal phosphate groups as ligands or substrates of Maf. A further clue comes from the observation that the structure of the Maf monomer bears similarity to that of the recently reported Methanococcus jannaschii Mj0226 protein. Just as for Maf, the structure of this predicted NTPase was determined as part of a structural genomics pilot project. The structural relation between Maf and Mj0226 was not apparent from sequence analysis approaches. These results emphasize the potential of structural genomics to reveal new unexpected connections between protein families previously considered unrelated. PMID- 10841542 TI - Analysis of mutations and suppressors affecting interactions between the subunits of the HIV type 1 reverse transcriptase. AB - HIV-1 reverse transcriptase (RT) catalyzes the conversion of genomic RNA into cDNA. The enzyme is a heterodimer of p66 and p51 subunits, and the dimerization of these subunits is required for optimal enzyme activity. To analyze this process at the genetic level, we developed constructs that permit the detection of the interaction between these subunits in the yeast two-hybrid system. Genetic analysis of RT subdomains required for heterodimerization revealed that the fingers and palm of p66 were dispensable for p51 interaction. However, as little as a 26-amino acid deletion at the C terminus of p51 prevented dimerization with p66. A primer grip mutation, L234A, previously shown to inhibit RT dimerization by biochemical assays, also prevented RT dimerization in the yeast two-hybrid system. Second-site mutations that restored RT dimerization in yeast to the L234A parent were recovered in the tryptophan repeat region at the dimer interface and at the polymerase active site, suggesting the involvement of these sites in RT dimerization. In vitro binding experiments confirmed the effects of the L234A mutation and the suppressor mutations on the interaction of the two subunits. The RT two-hybrid assay should facilitate the extensive genetic analysis of RT dimerization and should make possible the rapid screening of potential inhibitors of this essential process. PMID- 10841543 TI - Nutrient regulation of gene expression by the sterol regulatory element binding proteins: increased recruitment of gene-specific coregulatory factors and selective hyperacetylation of histone H3 in vivo. AB - We have evaluated the mechanism for sterol-regulated gene expression by the sterol regulatory element binding proteins (SREBPs) in intact cells. We show that activation of SREBPs by sterol depletion results in the increased binding of Sp1 to a site adjacent to SREBP in the promoter for the low density lipoprotein (LDL) receptor gene in vivo. Similarly, sterol depletion resulted in the increased recruitment of two distinct SREBP coregulatory factors, NF-Y and CREB, to the promoter for hydroxymethyl glutaryl CoA reductase, another key gene of intracellular cholesterol homeostasis. Furthermore, increased acetylation of histone H3 but not H4 was also detected in chromatin from both promoters on SREBP activation. Thus, SREBP activation results in the similar selective recruitment of different coregulatory generic transcription factors to two separate cholesterol-regulated promoters. These studies demonstrate the utility of the chromatin immunoprecipitation technique for analyzing the differential action of low-abundance transcription factors in fundamental regulatory events in intact cells. Our results also provide key in vivo support for the mechanism proposed from cell-free experiments, where SREBP increased the binding of Sp1 to the LDL receptor promoter. Finally, our findings also indicate that subtle differences in the pattern of core histone acetylation play a role in selective gene activation. PMID- 10841544 TI - Molecular modeling and in vitro activity of an HIV-1-encoded glutathione peroxidase. AB - Based on theoretical evidence, it has been proposed that HIV-1 may encode several selenoprotein modules, one of which (overlapping the env gp41-coding region) has highly significant sequence similarity to the mammalian selenoprotein glutathione peroxidase (GPx; EC ). The similarity score of the putative HIV-1 viral GPx homolog relative to an aligned set of known GPx is 6.3 SD higher than expected for random sequences of similar composition. Based on that alignment, a molecular model of the HIV-1 GPx was constructed by homology modeling from the bovine GPx crystal structure. Despite extensive truncation relative to the cellular GPx gene, the structural core and the geometry of the catalytic triad of selenocysteine, glutamine, and tryptophan are well conserved in the viral GPx. All of the insertions and deletions predicted by the alignment proved to be structurally feasible. The model is energetically favorable, with a computed molecular mechanics strain energy close to that of the bovine GPx structure, when normalized on a per-residue basis. However, considering the remote homology, this model is intended only to provide a working hypothesis allowing for a similar active site and structural core. To validate the theoretical predictions, we cloned the hypothetical HIV-1 gene and found it to encode functional GPx activity when expressed as a selenoprotein in mammalian cells. In transfected canine kidney cells, the increase in GPx activity ranged from 21% to 43% relative to controls (average 30%, n = 9, P < 0.0001), whereas, in transfected MCF7 cells, which have low endogenous GPx activity, a near 100% increase was observed (average 99%, n = 3, P < 0.05). PMID- 10841545 TI - Catalytic role of enzymes: short strong H-bond-induced partial proton shuttles and charge redistributions. AB - A two-step reaction mechanism (catalyzed alternatively by acid and base) with partial proton shuttles and charge redistributions promoted by short strong H bonds (SSHBs) (playing a dual role as an amphi-acid/base catalyst) is proposed to explain the enormous rate enhancement observed in enzymatic reactions involving carbanion intermediates. The SSHBs in the two-step reactions are found to be responsible for enhancing enzyme-substrate interactions in favor of the transition state structure over that of reactant. The detailed quantum theoretical studies of ketosteroid isomerase provide evidence of assisting roles of SSHB in enzymatic activity. The understanding of the two-step reaction mechanism would be a useful aid in designing novel functional enzymes and abzymes. PMID- 10841546 TI - Differential expression and function of members of the zfh-1 family of zinc finger/homeodomain repressors. AB - zfh-1 is a zinc finger/homeodomain transcriptional repressor in Drosophila that regulates differentiation of muscle and gonadal cells and is also expressed in the central nervous system (CNS). Binding sites for zfh-1 overlap with those for snail, and like snail, it recruits the corepressor CtBP-1. The protein ZEB-1 appears to be a vertebrate homologue of zfh-1 and is expressed in several tissues including muscle, CNS, and T lymphocytes, and during skeletal differentiation. Mutation of the ZEB-1 gene led to a severe T cell phenotype and skeletal defects but, interestingly, no defects were evident in other ZEB-1-expressing tissues. These results suggested that another ZEB-1-related factor may compensate for the loss of ZEB-1 in other tissues. Here, we characterize such a ZEB-1-related protein, which we have termed as ZEB-2. The overall organization of ZEB-2 is similar to ZEB-1 and zfh-1 and it has similar biochemical properties: it binds E boxes and interacts with CtBP-1 to repress transcription. However, there are also differences between ZEB-1 and ZEB-2, both in activity and tissue distribution. Whereas ZEB-1 and ZEB-2 overlap in skeletal muscle and CNS (providing an explanation for why mutation of ZEB-1 alone has little effect in these tissues), they show a different pattern of expression in lymphoid cells. ZEB-1, but not ZEB 2, is expressed in T cells from the thymus ZEB-2 appears to be expressed on splenic B cells. Additionally, ZEB-2 inhibits a wider spectrum of transcription factors than ZEB-1. PMID- 10841547 TI - Cell cycle inhibition by the anti-angiogenic agent TNP-470 is mediated by p53 and p21WAF1/CIP1. AB - Angiogenesis has been demonstrated to be essential for tumor growth and metastasis, and inhibition of angiogenesis is emerging as a promising strategy for treating cancer. Among the most potent inhibitors of angiogenesis is the fumagillin family of natural products. An analog of fumagillin, known as TNP-470 or AGM-1470, has been undergoing clinical trials for treating a variety of cancers. TNP-470 has been shown to block endothelial cell cycle progression in the late G(1) phase. Although the direct molecular target for TNP-470 has been identified as the type 2 methionine aminopeptidase (MetAP2), how inhibition of this enzyme leads to cell cycle arrest has remained unclear. We report that treatment of endothelial and other drug-sensitive cell types leads to the activation of the p53 pathway, causing an accumulation of the G(1) cyclin dependent kinase inhibitor p21(WAF1/CIP1). The requirement of p53 and p21(WAF1/CIP1) for the cell cycle inhibition by TNP-470 is underscored by the observation that cells deficient in p53 and p21(WAF1/CIP1) are resistant to TNP 470. These results shed significant light on the mechanism of cell cycle inhibition by TNP-470 and suggest an alternative method of activating p53 in endothelial cells to halt angiogenesis and tumor progression. PMID- 10841548 TI - Expression of a nonmyristylated variant of the catalytic subunit of protein kinase A during male germ-cell development. AB - The catalytic subunits of protein kinase A are transcribed in all mouse tissues from two distinct genes that code for the Calpha and Cbeta isoforms. Alternative promoters exist for the Cbeta gene that are used in a tissue-specific fashion and give rise to variants that differ in their amino-terminal sequences. We have characterized an alternative promoter that is present in the first intron of the Calpha gene and is transcriptionally active in male germ cells. Transcription from this promoter is coincident with the appearance of pachytene spermatocytes and leads to a Calpha protein (Calpha2) that contains a distinctive 7 amino acid amino-terminus differing from the 14 amino acid amino-terminus of Calpha1. The Calpha2 protein does not contain the myristylation signal present on Calpha1 and migrates at a lower molecular weight on SDS/PAGE gels. By Western blotting, we estimate that most or all of the Calpha protein present in mature sperm is Calpha2. The amino-terminal sequence of Calpha2 is similar to that of ovine sperm C as previously reported [San Agustin, J. T., Leszyk, J. D., Nuwaysir, L. M. & Witman, G. B. (1998) J. Biol. Chem. 273, 24874-24883], and we show by cDNA cloning that human sperm also express a highly related Calpha2 homolog. The Calpha2 subunit forms holoenzymes with either RIIalpha or RIalpha, and both activate at the same concentration of cyclic nucleotide. Because protein kinase A is thought to play a pivotal role in sperm motility and capacitation, the distinctive biochemical properties of the unmyristylated Calpha2 may be essential for fertility in the male. PMID- 10841549 TI - The preaggregated state of an amyloidogenic protein: hydrostatic pressure converts native transthyretin into the amyloidogenic state. AB - Protein misfolding and aggregation cause several diseases, by mechanisms that are poorly understood. The formation of amyloid aggregates is the hallmark of most of these diseases. Here, the properties and formation of amyloidogenic intermediates of transthyretin (TTR) were investigated by the use of hydrostatic pressure and spectroscopic techniques. Native TTR tetramers (T(4)) were denatured by high pressure into a conformation that exposes tryptophan residues to the aqueous environment. This conformation was able to bind the hydrophobic probe bis-(8 anilinonaphthalene-1-sulfonate), indicating persistence of elements of secondary and tertiary structure. Lowering the temperature facilitated the pressure-induced denaturation of TTR, which suggests an important role of entropy in stabilizing the native protein. Gel filtration chromatography showed that after a cycle of compression-decompression at 1 degrees C, the main species present was a tetramer, with a small population of monomers. This tetramer, designated T(4)*, had a non-native conformation: it bound more bis-(8-anilinonaphthalene-1 sulfonate) than native T(4), was less stable under pressure, and on decompression formed aggregates under mild acidic conditions (pH 5-5.6). Our data show that hydrostatic pressure converts native tetramers of TTR into an altered state that shares properties with a previously described amyloidogenic intermediate, and it may be an intermediate that lies on the aggregation pathway. This "preaggregated" state, which we call T(4)*, provides insight into the question of how a correctly folded protein may degenerate into the aggregation pathway in amyloidogenic diseases. PMID- 10841550 TI - Biosynthesis of terpenoids: 4-diphosphocytidyl-2C-methyl-D-erythritol synthase of Arabidopsis thaliana. AB - A hypothetical gene with similarity to the ispD gene of Escherichia coli was cloned from Arabidopsis thaliana cDNA. The ORF of 909 bp specifies a protein of 302 amino acid residues. The cognate chromosomal gene consists of 2,071 bp and comprises 11 introns with a size range of 78-202 bp. A fragment comprising amino acid residues 76-302 was expressed in a recombinant E. coli strain. The protein was purified to homogeneity and was shown to catalyze the formation of 4 diphosphocytidyl-2C-methyl-d-erythritol from 2C-methyl-d-erythritol 4-phosphate with a specific activity of 67 micromol small middle dotmin(-1) mg(-1). The Michaelis constants for 4-diphosphocytidyl-2C-methyl-d-erythritol and CTP were 500 microM and 114 microM, respectively. PMID- 10841551 TI - Identification of the single-strand telomeric DNA binding domain of the Saccharomyces cerevisiae Cdc13 protein. AB - The CDC13 gene of Saccharomyces cerevisiae is required both to protect telomeric DNA and to ensure proper function of yeast telomerase in vivo. We have previously demonstrated that Cdc13p has a high affinity single-strand telomeric DNA binding activity, although the primary amino acid sequence of Cdc13p has no previously characterized DNA binding motifs. We report here mapping of the Cdc13 DNA binding domain by a combination of proteolysis mapping and deletion cloning. The DNA binding domain maps to residues 557-694 of the 924-amino acid Cdc13 polypeptide, within the most basic region of Cdc13p. A slightly larger version of this domain can be efficiently expressed in Escherichia coli as a soluble small protein, with DNA binding properties comparable to those of the full-length protein. A single amino acid missense mutation within this domain results in thermolabile DNA binding and conditional lethality in yeast, consistent with the prediction that DNA binding should be essential for CDC13 function. These results show that Cdc13p contains a discrete substructure responsible for DNA binding and should facilitate structural characterization of this telomere binding protein. PMID- 10841552 TI - Thiol-disulfide exchange is involved in the catalytic mechanism of peptide methionine sulfoxide reductase. AB - Peptide methionine sulfoxide reductase (MsrA; EC ) reverses the inactivation of many proteins due to the oxidation of critical methionine residues by reducing methionine sulfoxide, Met(O), to methionine. MsrA activity is independent of bound metal and cofactors but does require reducing equivalents from either DTT or a thioredoxin-regenerating system. In an effort to understand these observations, the four cysteine residues of bovine MsrA were mutated to serine in a series of permutations. An analysis of the enzymatic activity of the variants and their free sulfhydryl states by mass spectrometry revealed that thiol disulfide exchange occurs during catalysis. In particular, the strictly conserved Cys-72 was found to be essential for activity and could form disulfide bonds, only upon incubation with substrate, with either Cys-218 or Cys-227, located at the C terminus. The significantly decreased activity of the Cys-218 and Cys-227 variants in the presence of thioredoxin suggested that these residues shuttle reducing equivalents from thioredoxin to the active site. A reaction mechanism based on the known reactivities of thiols with sulfoxides and the available data for MsrA was formulated. In this scheme, Cys-72 acts as a nucleophile and attacks the sulfur atom of the sulfoxide moiety, leading to the formation of a covalent, tetracoordinate intermediate. Collapse of the intermediate is facilitated by proton transfer and the concomitant attack of Cys-218 on Cys-72, leading to the formation of a disulfide bond. The active site is returned to the reduced state for another round of catalysis by a series of thiol-disulfide exchange reactions via Cys-227, DTT, or thioredoxin. PMID- 10841553 TI - Cockroach diuretic hormones: characterization of a calcitonin-like peptide in insects. AB - Insect diuretic hormones are crucial for control of water balance. We isolated from the cockroach Diploptera punctata two diuretic hormones (DH), Dippu-DH(31) and Dippu-DH(46), which increase cAMP production and fluid secretion in Malpighian tubules of several insect species. Dippu-DH(31) and -DH(46) contain 31 and 46 amino acids, respectively. Dippu-DH(46) belongs to the corticotropin releasing factor (CRF)-like insect DH family, whereas Dippu-DH(31) has little sequence similarity to the CRF-like DH, but is similar to the calcitonin family. Dippu-DH(46) and -DH(31) have synergistic effects in D. punctata but have only additive effects in Locusta migratoria. Dippu-DH(31) represents a distinct type of insect DH with actions that differ from those of previously identified insect peptides with diuretic activity. PMID- 10841554 TI - Investigation of routes and funnels in protein folding by free energy functional methods. AB - We use a free energy functional theory to elucidate general properties of heterogeneously ordering, fast folding proteins, and we test our conclusions with lattice simulations. We find that both structural and energetic heterogeneity can lower the free energy barrier to folding. Correlating stronger contact energies with entropically likely contacts of a given native structure lowers the barrier, and anticorrelating the energies has the reverse effect. Designing in relatively mild energetic heterogeneity can eliminate the barrier completely at the transition temperature. Sequences with native energies tuned to fold uniformly, as well as sequences tuned to fold reliably by a single or a few routes, are rare. Sequences with weak native energetic heterogeneity are more common; their folding kinetics is more strongly determined by properties of the native structure. Sequences with different distributions of stability throughout the protein may still be good folders to the same structure. A measure of folding route narrowness is introduced that correlates with rate and that can give information about the intrinsic biases in ordering arising from native topology. This theoretical framework allows us to investigate systematically the coupled effects of energy and topology in protein folding and to interpret recent experiments that investigate these effects. PMID- 10841556 TI - Defense by foot adhesion in a beetle (Hemisphaerota cyanea). AB - The beetle Hemisphaerota cyanea (Chrysomelidae; Cassidinae) responds to disturbance by activating a tarsal adhesion mechanism by which it secures a hold on the substrate. Its tarsi are oversized and collectively bear some 60,000 adhesive bristles, each with two terminal pads. While walking, the beetle commits but a small fraction of the bristles to contact with the substrate. But when assaulted, it presses its tarsi flatly down, thereby touching ground with all or nearly all of the bristles. Once so adhered, it can withstand pulling forces of up to 0.8 g ( approximately 60 times its body mass) for 2 min, and of higher magnitudes, up to >3 g, for shorter periods. Adhesion is secured by a liquid, most probably an oil. By adhering, the beetle is able to thwart attacking ants, given that it is able to cling more persistently than the ant persists in its assault. One predator, the reduviid Arilus cristatus, is able to feed on the beetle, possibly because by injecting venom it prevents the beetle from maintaining its tarsal hold. PMID- 10841555 TI - Mobilization of stem/progenitor cells by sulfated polysaccharides does not require selectin presence. AB - Employing carbohydrate ligands, which have been extensively used to block selectin function in vitro and in vivo, we have examined the involvement of such ligands in stem/progenitor cell mobilization in mice and monkeys. We found that sulfated fucans, branched and linear, are capable of increasing mature white cells in the periphery and mobilizing stem/progenitor cells of all classes (up to 32-fold) within a few hours posttreatment in a dose-dependent manner. To elicit the effect, the presence of sulfate groups was necessary, yet not sufficient, as certain sulfated hexosamines tested (chondroitin sulfates A or B) were ineffective. Significant mobilization of stem/progenitor cells and leukocytosis was elicited in selectin-deficient mice (L(-/-), PE(-/-), or LPE(-/-)) similar to that of wild-type controls, suggesting that the mode of action of sulfated fucans is not through blockade of known selectins. Other mechanisms have been entertained, in particular, the release of chemokines/cytokines, including some previously implicated in mobilization. Significant increases were documented in the levels of seven circulating chemokines/cytokines within a few hours after fucan sulfate treatment and support such a proposition. Additionally, an increase was noted in plasma metalloproteinase (MMP) 9, which might independently contribute to the mobilization process by enzymatically facilitating chemokine/cytokine release. Mobilization by sulfated polysaccharides provides a distinct paradigm in the mobilization process and uncovers an additional novel in vivo biological role for sulfated glycans. As similarly sulfated compounds were ineffective in vivo, the data also underscore the fact that polysaccharides with similar structures may elicit diverse in vivo effects. PMID- 10841557 TI - Olenid trilobites: the oldest known chemoautotrophic symbionts? AB - Late Cambrian to early Ordovician trilobites, the family Olenidae, were tolerant of oxygen-poor, sulfur-rich sea floor conditions, and a case is made that they were chemoautotrophic symbionts. Olenids were uniquely adapted to this habitat in the Lower Paleozoic, which was widespread in the Late Cambrian over Scandinavia. This life habit explains distinctive aspects of olenid morphology: wide thoraces and large numbers of thoracic segments, thin cuticle and, in some species, degenerate hypostome, and the occasional development of brood pouches. Geochemical and field evidence is consistent with this interpretation. Olenids occupied their specialized habitat for 60 million years until their extinction at the end of the Ordovician. PMID- 10841558 TI - Convergent adaptive radiations in Madagascan and Asian ranid frogs reveal covariation between larval and adult traits. AB - Recent studies have reported that independent adaptive radiations can lead to identical ecomorphs. Our phylogenetic analyses of nuclear and mitochondrial DNA sequences here indicate that a major radiation of ranid frogs on Madagascar produced morphological, physiological, and developmental characters that are remarkably similar to those that independently evolved on the Indian subcontinent. We demonstrate further that, in several cases, adult and larval stages each evolved sets of characters which are not only convergent between independent lineages, but also allowed both developmental stages to invade the same adaptive zone. It is likely that such covariations are produced by similar selective pressures on independent larval and adult characters rather than by genetic or functional linkage. We briefly discuss why larval/adult covariations might constitute an important evolutionary phenomenon in species for which more than one developmental stage potentially has access to multiple environmental conditions. PMID- 10841559 TI - Adaptive variation in lactate dehydrogenase-B gene expression: role of a stress responsive regulatory element. AB - Although changes in gene regulation may play an important role in adaptive evolution, there have been few attempts to investigate the molecular mechanisms responsible for adaptively significant variation in gene expression. Here we describe the mechanism underlying an adaptive difference in the expression of the lactate dehydrogenase-B gene (Ldh-B) between northern and southern populations of the fish Fundulus heteroclitus. Ldh-B regulatory sequences from northern and southern individuals, coupled to a luciferase reporter gene, were introduced into the livers of live fish. Deletion studies indicated that sequence changes between 400 and 500 bp upstream of the transcription start site resulted in a 2-fold difference in reporter gene transcription. These sequence changes can account for the previously observed 2-fold difference in Ldh-B transcription between populations. Variation in transcription factors did not play an important role. Sequences within the functionally important region resemble a mammary tumor virus glucocorticoid responsive element (MTV-GRE) in southern alleles, whereas northern alleles differ from the consensus by 1 bp. To test the hypothesis that this element is involved in the variation between populations of F. heteroclitus, we exposed transiently transgenic fish containing Ldh-B regulatory sequence/reporter gene constructs to handling stress or injected cortisol. Both treatments increased reporter gene transcription driven by southern alleles but not northern alleles, as expected if an MTV-GRE sequence were involved. This finding suggests that sequence variation in a GRE is the cause of the adaptive differences in Ldh B gene expression between populations and demonstrates that small changes in gene regulatory sequences can have important evolutionary consequences. PMID- 10841560 TI - Identification of phylogenetic footprints in primate tumor necrosis factor-alpha promoters. AB - The human tumor necrosis factor-alpha (TNF-alpha) gene encodes a pleiotropic cytokine that plays a critical role in basic immunologic processes. To investigate the TNF-alpha regulatory region in the primate lineage, we isolated TNF-alpha promoters from representative great apes, Old World monkeys, and New World monkeys. We demonstrate that there is a nonuniform distribution of fixed human differences in the TNF-alpha promoter. We define a "fixed human difference" as a site that is not polymorphic in humans, but which differs in at least one of the seven primate sequences examined. Furthermore, we identify two human TNF alpha promoter single nucleotide polymorphisms that are putative ancestral polymorphisms, because each of the human polymorphic nucleotides was found at the identical site in at least one of the other primate sequences. Strikingly, the largest conserved region among the primate species, a 69-nt "phylogenetic footprint," corresponds to a region of the human TNF-alpha promoter that forms the transcriptionally active nucleoprotein-DNA complex, essential for gene regulation. By contrast, other regions of the TNF-alpha promoter, which exhibit a high density of variable sites, are nonessential for gene expression, indicating that distinct TNF-alpha promoter regions have been subjected to different evolutionary constraints depending on their function. TNF-alpha is the first case in which a promoter region dissected by functional analyses can be correlated with nucleotide polymorphism and variability in primate lineages. The results suggest that patterns of polymorphism and divergence are likely to be useful in identifying candidate regions important for gene regulation in other immune response genes. PMID- 10841561 TI - Anopheles gambiae pilot gene discovery project: identification of mosquito innate immunity genes from expressed sequence tags generated from immune-competent cell lines. AB - Together with AIDS and tuberculosis, malaria is at the top of the list of devastating infectious diseases. However, molecular genetic studies of its major vector, Anopheles gambiae, are still quite limited. We have conducted a pilot gene discovery project to accelerate progress in the molecular analysis of vector biology, with emphasis on the mosquito's antimalarial immune defense. A total of 5,925 expressed sequence tags were determined from normalized cDNA libraries derived from immune-responsive hemocyte-like cell lines. The 3,242 expressed sequence tag-containing cDNA clones were grouped into 2,380 clone clusters, potentially representing unique genes. Of these, 1,118 showed similarities to known genes from other organisms, but only 27 were identical to previously known mosquito genes. We identified 38 candidate genes, based on sequence similarity, that may be implicated in immune reactions including antimalarial defense; 19 of these were shown experimentally to be inducible by bacterial challenge, lending support to their proposed involvement in mosquito immunity. PMID- 10841562 TI - Molecular evidence for a relationship between LINE-1 elements and X chromosome inactivation: the Lyon repeat hypothesis. AB - X inactivation is a chromosome-specific form of genetic regulation in which thousands of genes on one homologue become silenced early in female embryogenesis. Although many aspects of X inactivation are now understood, the spread of the X inactivation signal along the entire length of the chromosome remains enigmatic. Extending the Gartler-Riggs model [Gartler, S. M. & Riggs, A. D. (1983) Annu. Rev. Genet. 17, 155-190], Lyon recently proposed [Lyon, M. F. (1998) Cytogenet. Cell Genet. 80, 133-137] that a nonrandom organization of long interspersed element (LINE) repetitive sequences on the X chromosome might be responsible for its facultative heterochromatization. In this paper, we present data indicating that the LINE-1 (L1) composition of the human X chromosome is fundamentally distinct from that of human autosomes. The X chromosome is enriched 2-fold for L1 repetitive elements, with the greatest enrichment observed for a restricted subset of LINE-1 elements that were active <100 million years ago. Regional analysis of the X chromosome reveals that the most significant clustering of these elements is in Xq13-Xq21 (the center of X inactivation). Genomic segments harboring genes that escape inactivation are significantly reduced in L1 content compared with X chromosome segments containing genes subject to X inactivation, providing further support for the association between X inactivation and L1 content. These nonrandom properties of L1 distribution on the X chromosome provide strong evidence that L1 elements may serve as DNA signals to propagate X inactivation along the chromosome. PMID- 10841563 TI - A phylogenetically conserved NAD+-dependent protein deacetylase activity in the Sir2 protein family. AB - The yeast Sir2 protein, required for transcriptional silencing, has an NAD(+) dependent histone deacetylase (HDA) activity. Yeast extracts contain a NAD(+) dependent HDA activity that is eliminated in a yeast strain from which SIR2 and its four homologs have been deleted. This HDA activity is also displayed by purified yeast Sir2p and homologous Archaeal, eubacterial, and human proteins, and depends completely on NAD(+) in all species tested. The yeast NPT1 gene, encoding an important NAD(+) synthesis enzyme, is required for rDNA and telomeric silencing and contributes to silencing of the HM loci. Null mutants in this gene have significantly reduced intracellular NAD(+) concentrations and have phenotypes similar to sir2 null mutants. Surprisingly, yeast from which all five SIR2 homologs have been deleted have relatively normal bulk histone acetylation levels. The evolutionary conservation of this regulated activity suggests that the Sir2 protein family represents a set of effector proteins in an evolutionarily conserved signal transduction pathway that monitors cellular energy and redox states. PMID- 10841564 TI - Recombinase-activating gene (RAG) 2-mediated V(D)J recombination is not essential for tumorigenesis in Atm-deficient mice. AB - The majority of Atm-deficient mice die of malignant thymic lymphoma by 4-5 mo of age. Cytogenetic abnormalities in these tumors are consistently identified within the Tcr alpha/delta locus, suggesting that tumorigenesis is secondary to aberrant responses to double-stranded DNA breaks that occur during V(D)J recombination. Since V(D)J recombination is a recombinase-activating gene (RAG)-dependent process, we generated Rag2(-/-)Atm(-/-) mice to assess the requirement for RAG dependent recombination in thymic lymphomagenesis. In contrast to expectation, the data presented here indicate that development of malignant thymic lymphoma in Atm(-/-) mice is not prevented by loss of RAG-2 and thus is not dependent on V(D)J recombination. Malignant thymic lymphomas in Rag2(-/-)Atm(-/-) mice occurred at a lower frequency and with a longer latency as compared with Atm(-/-) mice. Importantly, cytogenetic analysis of these tumors indicated that multiple chromosomal abnormalities occurred in each tumor, but that none of these involved the Tcr alpha/delta locus. Nonmalignant peripheral T cells from TCR-transgenic Rag2(-/-)Atm(-/-) mice also revealed a substantial increase in translocation frequency, suggesting that these translocations are early events in the process of tumorigenesis. These data are consistent with the hypothesis that the major mechanism of tumorigenesis in Atm(-/-) mice is via chromosomal translocations and other abnormalities that are secondary to aberrant responses to double-stranded DNA breaks. Furthermore, these data suggest that V(D)J recombination is a critical, but not essential, event during which Atm-deficient thymocytes are susceptible to developing chromosome aberrations that predispose to malignant transformation. PMID- 10841565 TI - Genetic reconstitution of systemic lupus erythematosus immunopathology with polycongenic murine strains. AB - We previously produced three congenic strains carrying lupus susceptibility genes (Sle1-Sle3) from the lupus-prone NZM2410 mouse on the C57BL/6 background and characterized their component phenotypes. Sle1 mediates the loss of tolerance to nuclear antigens; Sle2 lowers the activation threshold of B cells; and Sle3 mediates a dysregulation of CD4(+) T cells. We have now created a collection of bi- and tricongenic strains with these intervals and assessed the autoimmune phenotypes they elicit in various combinations. Our results indicate that Sle1 is key for the development of fatal lupus. The combination of Sle1 with Sle2, Sle3, or the BXSB-derived autoimmune accelerating gene yaa results in the development of systemic autoimmunity with variably penetrant severe glomerulonephritis culminating in kidney failure. In contrast, two locus combinations of Sle2, Sle3, and yaa failed to mediate fatal disease. These results indicate that the loss of tolerance to chromatin mediated by Sle1 is essential for disease pathogenesis and identify the pathway occupied by Sle1 as a strategic target for therapeutic intervention in systemic lupus erythematosus. The coexpression of Sle1, Sle2, and Sle3 as a B6-triple congenic results in severe systemic autoimmunity and fully penetrant, fatal glomerulonephritis. These results demonstrate the fulfillment of the genetic equivalent of Koch's postulate, where susceptibility loci in a lupus prone strain have been identified by a genome scan, isolated and functionally characterized by congenic dissection, and finally shown to mediate full disease expression when recombined in a normal genome. PMID- 10841566 TI - A critical role for eotaxin in experimental oral antigen-induced eosinophilic gastrointestinal allergy. AB - Despite marked advances in the understanding of allergic responses, the mechanisms regulating gastrointestinal allergy are not very well understood. We have developed a model of antigen-induced eosinophil-associated gastrointestinal allergy and characterized the role of eotaxin and IL-5. Challenge of allergen sensitized mice with oral allergen, in the form of enteric-coated beads, resulted in marked allergen-specific IgG(1) and IgE, Th(2)-type (IL-4 and IL-5) cytokine production, and eosinophil accumulation in the blood and small intestine. In the genetic absence of eotaxin, a chemokine constitutively expressed in the gastrointestinal tract, eosinophil recruitment into the small intestine was ablated, and these mice developed enhanced eosinophil accumulation in the blood compared with wild-type mice. Interestingly, in the absence of IL-5, allergen challenge promoted partial eosinophil accumulation into the small intestine and a decline in circulating eosinophil levels. Collectively, these results establish that the accumulation of gastrointestinal eosinophils is antigen induced, can occur independent of IL-5, and provides a molecular mechanism to explain the dichotomy between peripheral blood and tissue eosinophilia. Furthermore, eotaxin is identified as a critical regulator of antigen-induced eosinophilic inflammation in the gastrointestinal tract. PMID- 10841567 TI - Delayed-type hypersensitivity to Phlebotomus papatasi sand fly bite: An adaptive response induced by the fly? AB - The saliva of bloodsucking arthropods contains a large array of pharmacologically active compounds that assist hematophagy. Arthropod saliva is also responsible for causing uncomfortable allergic responses in its vertebrate hosts. In this article, we investigate whether the sand fly Phlebotomus papatasi, known to produce a strong delayed-type hypersensitivity (DTH) in humans, could benefit from, and possibly adaptively induce, this response in their vertebrate hosts. In this study, we show that flies fed on humans to completion nearly twice as fast in DTH sites as compared with normal skin sites. DTH sites had significantly larger blood flow as measured by the laser Doppler method. Sand flies feeding at sites in mouse ears that had a DTH response also fed faster than at normal sites. We conclude that in the case of P. papatasi, and possibly other arthropods such as fleas and bed bugs, the strong saliva-induced DTH response may reflect an adaptation of the fly to manipulate host immunity for the insect's own advantage. PMID- 10841568 TI - Trans-splicing vectors expand the utility of adeno-associated virus for gene therapy. AB - Adeno-associated viral (AAV) vectors have demonstrated considerable promise for gene therapy of inherited diseases. However, with a packaging size of <5 kb, applications have been limited to relatively small disease genes. Based on the finding that AAV genomes undergo intermolecular circular concatamerization after transduction in muscle, we have developed a paradigm to increase the size of delivered transgenes with this vector through trans-splicing between two independent vectors coadministered to the same tissue. When two vectors encoding either the 5' or 3' portions of the erythropoietin genomic locus were used, functional erythropoietin protein was expressed in muscle subsequent to the formation of intermolecular circular concatamers in a head-to-tail orientation through trans-splicing between these two independent vector genomes. These findings will allow for the application of AAV technologies to a wider variety of diseases for which therapeutic transgenes exceed the packaging limitation of present AAV vectors. PMID- 10841569 TI - Perlecan is required to inhibit thrombosis after deep vascular injury and contributes to endothelial cell-mediated inhibition of intimal hyperplasia. AB - Perlecan, a heparan sulfate proteoglycan, has been suggested to be critical for regulation of vascular repair. We generated clones of endothelial cells expressing an antisense vector targeting domain III of perlecan. Transfected cells produced significantly less perlecan than parent cells and showed a reduced ability to inhibit the binding and mitogenic activity of fibroblast growth factor 2 in vascular smooth muscle cells. Endothelial cells were seeded onto three dimensional polymeric matrices and implanted adjacent to porcine carotid arteries subjected to deep injury. Although the parent endothelial cells prevented occlusive thrombosis, perlecan-deficient cells were completely ineffective. The ability of endothelial cells to inhibit intimal hyperplasia, however, was abrogated only in part by perlecan suppression. The differential regulation by perlecan of these different aspects of vascular repair may explain why control of clinical clot formation does not lead to full control of intimal hyperplasia. Thus the use of genetically modified tissue-engineered cells provides a new approach for dissecting the role of specific factors within the complex environment of the blood vessel wall. PMID- 10841570 TI - Extensive neurite outgrowth and active synapse formation on self-assembling peptide scaffolds. AB - A new type of self-assembling peptide (sapeptide) scaffolds that serve as substrates for neurite outgrowth and synapse formation is described. These peptide-based scaffolds are amenable to molecular design by using chemical or biotechnological syntheses. They can be tailored to a variety of applications. The sapeptide scaffolds are formed through the spontaneous assembly of ionic self complementary beta-sheet oligopeptides under physiological conditions, producing a hydrogel material. The scaffolds can support neuronal cell attachment and differentiation as well as extensive neurite outgrowth. Furthermore, they are permissive substrates for functional synapse formation between the attached neurons. That primary rat neurons form active synapses on such scaffold surfaces in situ suggests these scaffolds could be useful for tissue engineering applications. The buoyant sapeptide scaffolds with attached cells in culture can be transported readily from one environment to another. Furthermore, these peptides did not elicit a measurable immune response or tissue inflammation when introduced into animals. These biological materials created through molecular design and self assembly may be developed as a biologically compatible scaffold for tissue repair and tissue engineering. PMID- 10841571 TI - Absence of muscarinic cholinergic airway responses in mice deficient in the cyclic nucleotide phosphodiesterase PDE4D. AB - Muscarinic cholinergic signaling plays an essential role in the control of the normal airway functions and in the development of pulmonary pathologies including asthma. In this paper we demonstrate that the airways of mice deficient in a cAMP specific phosphodiesterase (PDE4D) are no longer responsive to cholinergic stimulation. Airway hyperreactivity that follows exposure to antigen was also abolished in PDE4D(-/-) mice, despite an apparently normal lung inflammatory infiltration. The loss of cholinergic responsiveness was specific to the airway, not observed in the heart, and was associated with a loss of signaling through muscarinic receptors with an inability to decrease cAMP accumulation. These findings demonstrate that the PDE4D gene plays an essential role in cAMP homeostasis and cholinergic stimulation of the airway, and in the development of hyperreactivity. In view of the therapeutic potentials of PDE4 inhibitors, our findings provide the rationale for novel strategies that target a single PDE isoenzyme. PMID- 10841572 TI - Ecteinascidin 743, a transcription-targeted chemotherapeutic that inhibits MDR1 activation. AB - Ecteinascidin 743 (ET-743), a highly promising marine-based antitumor agent presently in phase II clinical trials, has been shown to interfere with the binding of minor-groove-interacting transcription factors, particularly NF-Y, with their cognate promoter elements in vitro. We have shown that NF-Y is a central mediator of activation of transcription of the human P glycoprotein gene (MDR1) by a variety of inducers and that NF-Y functions by recruiting the histone acetyltransferase PCAF to the MDR1 promoter. In the present study, we tested whether ET-743 could block activation of the MDR1 promoter by agents that mediate their effect through the NF-Y/PCAF complex. We report that physiologically relevant concentrations of ET-743 abrogate transcriptional activation of both the endogenous MDR1 gene and MDR1 reporter constructs by the histone deacetylase inhibitors as well as by UV light, with minimal effect on constitutive MDR1 transcription. Notably, this inhibition does not alter the promoter-associated histone hyperacetylation induced by histone deacetylase inhibitors, suggesting an in vivo molecular target downstream of NF-Y/PCAF binding. ET-743 is therefore the prototype for a distinct class of transcription-targeted chemotherapeutic agents and may be an efficacious adjuvant to the treatment of multidrug-resistant tumors. PMID- 10841573 TI - Interference of transcriptional activation by the antineoplastic drug ecteinascidin-743. AB - Ecteinascidin-743 (ET-743) is a tetrahydroisoquinoline alkaloid isolated from the tunicate Ecteinascidia turbinata currently under phase II clinical trials for its potent anticancer activity. ET-743 binds DNA in the minor groove and forms covalent adducts with some sequence specificity. It selectively inhibits in vitro binding of the CCAAT box factor NF-Y. In this study, we assayed ET-743 function in vivo on the HSP70 promoter. On heat induction, the drug blocks transcription rapidly at pharmacological concentrations and in a CCAAT-dependent manner, whereas the activity of the CCAAT-less simian virus 40 promoter is not affected. The effect is exerted at the mRNA level. The distamycin-like alkylating tallimustine is inactive in these assays. Binding of NF-Y and of the heat-shock factor is normal in ET-743-treated cells. Run-on analysis of several endogenous genes further proves that the drug has rapid, profound, and selective negative effects on transcription. Thus, this marine-derived compound is a promoter specific, transcription-interfering agent. PMID- 10841574 TI - In vitro correlates of HIV-2-mediated HIV-1 protection. AB - A prospective study of high-risk commercial sex workers in Senegal has shown that HIV-2 infection may reduce the risk of subsequent HIV-1 infection; these findings have been confirmed and extended, now with 13 years of observation. While exploring the biological mechanisms behind this natural protection, we found that a significant proportion of peripheral blood mononuclear cells obtained from HIV 2-infected subjects resisted in vitro challenge with CCR5-dependent HIV-1 viruses but not CXCR4-dependent viruses. High levels of beta-chemokines, the natural ligands of the CCR5 coreceptor, were correlated with low levels of viral replication, and resistance was abrogated by antibodies to beta-chemokines. Our results suggest that beta-chemokine-mediated resistance may be an important correlate of HIV protection against HIV-1 infection and relevant to HIV vaccine design. PMID- 10841575 TI - Intergeneric poliovirus recombinants for the treatment of malignant glioma. AB - Poliovirus neuropathogenicity depends on sequences within the 5' nontranslated region of the virus. Exchange of the poliovirus internal ribosomal entry site with its counterpart from human rhinovirus type 2 resulted in attenuation of neurovirulence in primates. Despite deficient virus propagation in cells of neuronal origin, nonpathogenic polio recombinants retain excellent growth characteristics in cell lines derived from glial neoplasms. Susceptibility of malignant glioma cells to poliovirus may be mediated by expression of a poliovirus receptor, CD155, in glial neoplasms. Intergeneric polio recombinants with heterologous internal ribosomal entry site elements unfolded strong oncolytic potential against experimentally induced gliomas in athymic mice. Our observations suggest that highly attenuated poliovirus recombinants may have applicability as biotherapeutic antineoplastic agents. PMID- 10841576 TI - Human monoclonal antibodies reactive to oligodendrocytes promote remyelination in a model of multiple sclerosis. AB - Promoting remyelination, a major goal of an effective treatment for demyelinating diseases, has the potential to protect vulnerable axons, increase conduction velocity, and improve neurologic deficits. Strategies to promote remyelination have focused on transplanting oligodendrocytes (OLs) or recruiting endogenous myelinating cells with trophic factors. Ig-based therapies, routinely used to treat a variety of neurological and autoimmune diseases, underlie our approach to enhance remyelination. We isolated two human mAbs directed against OL surface antigens that promoted significant remyelination in a virus-mediated model of multiple sclerosis. Four additional OL-binding human mAbs did not promote remyelination. Both human mAbs were as effective as human i.v. Ig, a treatment shown to have efficacy in multiple sclerosis, and bound to the surface of human OLs suggesting a direct effect of the mAbs on the cells responsible for myelination. Alternatively, targeting human mAbs to areas of central nervous system (CNS) pathology may facilitate the opsonization of myelin debris, allowing repair to proceed. Human mAbs were isolated from the sera of individuals with a form of monoclonal gammopathy. These individuals carry a high level of monoclonal protein in their blood without detriment, lending support to the belief that administration of these mAbs as a therapy would be safe. Our results are (i) consistent with the hypothesis that CNS-reactive mAbs, part of the normal Ig repertoire in humans, may help repair and protect the CNS from pathogenic immune injury, and (ii) further challenge the premise that Abs that bind OLs are necessarily pathogenic. PMID- 10841577 TI - Alzheimer-like neurodegeneration in aged antinerve growth factor transgenic mice. AB - Neurotrophin nerve growth factor (NGF) has been suggested to be involved in age related neurodegenerative diseases, but no transgenic model is currently available to study this concept. We have obtained transgenic mice expressing a neutralizing anti-NGF recombinant antibody, in which the levels of antibodies are three orders of magnitude higher in adult than in newborn mice [F.R., S. C. , A.C., E. Di Daniel, J. Franzot, S. Gonfloni, G. Rossi, N. B. & A. C. (2000) J. Neurosci., 20, 2589-2601]. In this paper, we analyze the phenotype of aged anti NGF transgenic mice and demonstrate that these mice acquire an age-dependent neurodegenerative pathology including amyloid plaques, insoluble and hyperphosphorylated tau, and neurofibrillary tangles in cortical and hippocampal neurons. Aged anti-NGF mice also display extensive neuronal loss throughout the cortex, cholinergic deficit in the basal forebrain, and behavioral deficits. The overall picture is strikingly reminiscent of human Alzheimer's disease. Aged anti NGF mice represent, to our knowledge, the most comprehensive animal model for this severe neurodegenerative disease. Also, these results demonstrate that, in mice, a deficit in the signaling and/or transport of NGF leads to neurodegeneration. PMID- 10841578 TI - Auditory processing parallels reading abilities in adults. AB - A broad battery of psychoacoustic measures and standard measures of reading and spelling were applied to 102 adults. The test group included individuals with a childhood history of reading difficulties and controls with no reported reading difficulties. Reading scores were variable in both groups. Poor auditory processing abilities were recorded in poor readers; particular difficulties were posed by tasks requiring spectral distinctions, the simplest of which was pure tone frequency discrimination. In absolute terms, the greatest deficits were recorded in tasks in which stimuli were presented in brief forms and in rapid succession. Auditory processing abilities accounted for more than 50% of the reading score variance in the control group, but their correlation with reading scores was lower in the group with childhood histories of reading difficulties. The additional variability in the latter group resulted largely from the prevalence of reading-compensated poor psychoacoustic performers, whose short term word memory was also typically poor. Taken together, these findings support a link between impaired auditory resolution and poor reading. Psychoacoustic difficulties are largely retained through adulthood and may be the source of the retained reading difficulties. PMID- 10841579 TI - Kainate receptor-mediated synaptic currents in cerebellar Golgi cells are not shaped by diffusion of glutamate. AB - We report the presence of kainate receptors (KARs) in cerebellar Golgi cells of wild-type but not GluR6-deficient mice. Parallel fiber stimulation activates KAR mediated synaptic currents [KAR-excitatory postsynaptic currents (EPSCs)] of small amplitude. KAR-EPSCs greatly differ from synaptic currents mediated by alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) receptors (AMPAR EPSCs) at the same synapse. KAR-EPSCs display slow rise and decay time and summate in response to a train of stimulations. By using PDA, a low-affinity competitive antagonist and agents that modify the clearance of glutamate, we show that these properties cannot be explained by diffusion of glutamate outside of the synaptic cleft and activation of extrasynaptic KARs. These data suggest that the slow kinetic of KAR-EPSCs is due to intrinsic properties of KARs being localized at postsynaptic sites. The contrasting properties of KAR- and AMPAR EPSCs in terms of kinetics and summation offer the possibility for a glutamatergic synapse to integrate excitatory inputs over two different time scales. PMID- 10841580 TI - Mouse numb is an essential gene involved in cortical neurogenesis. AB - During neurogenesis of the mammalian neocortex, neural progenitor cells divide to generate daughter cells that either become neurons or remain as progenitor cells. The mouse numb (m-numb) gene encodes a membrane-associated protein that is asymmetrically localized to the apical cell membrane of dividing cortical progenitor cells and may be segregated to only the apical daughter cell that has been suggested to remain as a progenitor cell. To examine m-numb function during neural development, we generated a loss-of-function mutant allele of m-numb. Mice homozygous for this mutation exhibit severe defects in cranial neural tube closure and precocious neuron production in the forebrain and die around embryonic day 11.5 (E11. 5). These findings suggest that m-numb is an essential gene that plays a role in promoting progenitor cell fate during cortical neurogenesis. PMID- 10841581 TI - A unique virus complex causes Ageratum yellow vein disease. AB - Ageratum conyzoides L., a weed species widely distributed throughout southeast Asia, frequently exhibits striking yellow vein symptoms associated with infection by Ageratum yellow vein virus (AYVV), a member of the Geminiviridae (genus Begomovirus). Most begomoviruses have bipartite genomes (DNAs A and B), but only a DNA A has been identified for AYVV. We demonstrate that yellow vein disease of A. conyzoides results from co-infection by AYVV DNA A (2,741 nt) and a circular DNA that is approximately half its size (1,347 nt) that we designate DNA beta. Apart from the sequence TAATATTAC, common to all geminiviruses and containing the initiation site of rolling circle replication, DNA beta shows negligible sequence homology either to AYVV DNA A or to DNA B associated with bipartite begomoviruses. DNA beta depends on DNA A for replication and is encapsidated by DNA A-encoded coat protein and so has characteristics of a DNA satellite. However, systemic infection of A. conyzoides by DNA A alone is sporadic and asymptomatic, and DNA A accumulation is reduced to 5% or less of its accumulation in the presence of DNA beta. Therefore, DNA A and DNA beta together form a previously unrecognized disease-inducing complex. Our data also demonstrate that the nanovirus-like DNA 1 component associated with infected A. conyzoides plays no essential role in the disease and represents a satellite-like DNA. Furthermore, the satellite DNA previously found associated with tomato leaf curl virus is probably a defective DNA beta homologue. PMID- 10841582 TI - Anaerobic chlorophyll isocyclic ring formation in Rhodobacter capsulatus requires a cobalamin cofactor. AB - The isocyclic ring of bacteriochlorophyll (BChl) is formed by the conversion of Mg-protoporphyrin monomethyl ester (MPE) to protochlorophyllide (PChlide). Similarities revealed by blast searches with the putative anaerobic MPE-cyclase BchE suggested to us that this protein also uses a cobalamin cofactor. We found that vitamin B(12) (B(12))-requiring mutants of the bluE and bluB genes of Rhodobacter capsulatus, grown without B(12), accumulated Mg-porphyrins. Laser desorption/ionization time-of-flight (LDI-TOF) MS and NMR spectroscopy identified them as MPE and its 3-vinyl-8-ethyl (mvMPE) derivative. An in vivo assay was devised for the cyclase converting MPE to PChlide. Cyclase activity in the B(12) dependent mutants required B(12) but not protein synthesis. The following reaction mechanism is proposed for this MPE-cyclase reaction. Adenosylcobalamin forms the adenosyl radical, which leads to withdrawal of a hydrogen atom and formation of the benzylic-type 13(1)-radical of MPE. Withdrawal of an electron gives the 13(1)-cation of MPE. Hydroxyl ion attack on the cation gives 13(1) hydroxy-MPE. Withdrawal of three hydrogen atoms leads successively to 13(1)-keto MPE, its 13(2)-radical, and cyclization to PChlide. PMID- 10841584 TI - Tobacco related disease research program. PMID- 10841583 TI - Changes in period mRNA levels in the brain and division of labor in honey bee colonies. AB - Previous research showed that age-related division of labor in honey bees is associated with changes in activity rhythms; young adult bees perform hive tasks with no daily rhythms, whereas older bees forage with strong daily rhythms. We report that this division of labor is also associated with differences in both circadian rhythms and mRNA levels of period, a gene well known for its role in circadian rhythms. The level of period mRNA in the brain oscillated in bees of all ages, but was significantly higher at all times in foragers. Elevated period mRNA levels cannot be attributed exclusively to aging, because bees induced to forage precociously because of a change in social environment had levels similar to normal age foragers. These results extend the regulation of a "clock gene" to a social context and suggest that there are connections at the molecular level between division of labor and chronobiology in social insects. PMID- 10841585 TI - Patterns of adolescent smoking initiation rates by ethnicity and sex. AB - OBJECTIVE: To define US national sex specific rates of smoking initiation among Hispanic, non-Hispanic white, and African American adolescents aged 12-17 years for each calendar year from 1940 through 1992. METHODS: Adult survey data from the tobacco use supplement of the Current Population Survey in 1992-93 and 1995 96 were used to reconstruct the age at which individuals began to smoke and the calendar year in which they were that age. From these data, the number of individuals who began a calendar year as never smokers and who were aged 12-17 years during that year could be estimated and formed the denominator of the initiation rate. The number of these individuals who reported taking up smoking during that year formed the numerator of the initiation rate. RESULTS: Initiation rates among male adolescents in each of the three racial/ethnic groups have declined since 1945. However, since 1983, initiation rates among male adolescents overall have increased. Non-Hispanic white male adolescents generally initiated cigarette smoking at higher rates than Hispanic or African American male adolescents. Initiation rates among Hispanic male adolescents have not been statistically different from initiation rates among African American male adolescents. From 1978 to 1982, initiation rates among Hispanic and African American male adolescents experienced a sharp decline, and the rate of decline was steeper than that experienced by non-Hispanic white male adolescents. Initiation rates among female adolescents have increased since 1940, catching up to male adolescent initiation rates by the mid 1970s. Initiation rates among female adolescents appeared to level off or increased slightly again from the mid 1980s to 1990. Non-Hispanic white female adolescents generally initiate cigarette smoking at higher rates than Hispanic or African American female adolescents. Initiation rates among non-Hispanic white and African American female adolescents equalled the initiation rates of their male counterparts by the mid 1970s, but initiation rates among Hispanic female adolescents did not overlap with initiation rates of Hispanic male adolescents until 1990. From 1975 to 1980, initiation rates among African American female adolescents decreased sharply, but, unlike initiation rates among the two other ethnic groups, rates continued to decline from 1984 to 1990. CONCLUSIONS: Different patterns of increasing and decreasing smoking initiation among sex and ethnic adolescent groups suggest the effect of varying social and cultural influences. These findings support the importance of including ethnic factors in studies of smoking behaviour. PMID- 10841586 TI - Ethnic differences in adolescent smoking prevalence in California: are multi ethnic youth at higher risk? AB - OBJECTIVE: Although ethnic differences in adolescent smoking have been well documented, smoking among multi-ethnic adolescents has received little research attention. This study examined smoking prevalence and tobacco related psychosocial risk factors among multi-ethnic adolescents in California, as compared with white, African American, Asian American, and Hispanic adolescents. DESIGN: This study used a cross sectional design. Data were obtained from the independent evaluation of the California Tobacco Control, Prevention, and Education Program. SETTING: Students completed a paper-and-pencil survey in their classrooms. SUBJECTS: Data were collected from a stratified random sample of 5072 eighth grade students (age 13-14 years) in California during the 1996-97 school year. The data were weighted by school enrollment in analyses to make the estimates representative of the population of California students attending public schools. MAIN OUTCOME MEASURES: Outcome variables included self reported smoking prevalence, susceptibility to smoking, access to tobacco, and related attitudes, beliefs, and behaviours. Ethnicity was assessed with a self reported, "check all that apply" question. RESULTS: Results indicated that multi-ethnic adolescents were at higher risk than single-ethnic adolescents on several variables, including 30 day cigarette smoking prevalence, lifetime smokeless tobacco use, buying cigarettes, receiving cigarette offers, and expected friends' reaction if the respondent smoked. For several other variables (lifetime cigarette smoking prevalence, susceptibility to smoking, and number of friends who smoked), Hispanic adolescents were at higher risk than all other ethnic groups including multi-ethnic adolescents. Although susceptibility to smoking was highest among Hispanics, multi-ethnic adolescents scored significantly higher on susceptibility than the three other single-ethnic groups. CONCLUSIONS: Multi ethnic adolescents may be at increased risk for smoking and may have easier access to cigarettes. Culturally targeted smoking prevention interventions for adolescents should address the unique social challenges faced by multi-ethnic adolescents that may increase their risk for smoking. PMID- 10841587 TI - Minors' access to tobacco before and after the California STAKE Act. AB - OBJECTIVE: To assess the effect of implementation and enforcement of the California STAKE Act on minors' access to tobacco by examining sales over time in the same stores. DESIGN: Sixteen year old girls and boys attempted to purchase cigarettes in the same 72 stores, in the same manner, in five time periods: August 1994 (before implementation of legislation); August 1995 (immediately after implementation); August 1996; March 1998; and January 1999 (all postimplementation). OUTCOME MEASURE: Percentage of successful cigarette purchases over time, in different ethnic communities. RESULTS: Minors' access rate decreased significantly from 41.2% before implementation of legislation (1994) to 12.7% after implementation (1998). The same stores were 3-5 times more likely to sell cigarettes to minors before than after the legislation was implemented, irrespective of ethnic census tract. CONCLUSIONS: The California STAKE Act and its enforcement have been effective in reducing minors' access to tobacco in all ethnic communities. PMID- 10841588 TI - Anti-smoking advertising campaigns targeting youth: case studies from USA and Canada. AB - OBJECTIVE: To assist in planning anti-smoking advertising that targets youth. Using five US state campaigns, one US research study, and a Canadian initiative as exemplars, an attempt is made to explain why certain advertising campaigns have been more cost effective than others in terms of reducing adolescent smoking prevalence. Several factors which prior research and theory suggest may be important to cost effectiveness are examined. Specifically, three variables pertaining to the advertising message (content, consistency, and clarity) and two variables related to the advertising execution or style (age of spokesperson and depiction of smoking behaviour) are studied. DESIGN: A case study approach has been combined with supplemental data collection and analysis. To assess campaign effects, published articles and surveys of adolescent smoking prevalence in campaign versus control (non-campaign) locations were utilised. Adolescent subjects provided supplemental data on the advertising message variables. Trained adults content analysed each advertisement to assess the executional variables. SUBJECTS: A total of 1128 seventh grade (age 12-13 years) and 10th grade (age 15 16 years) students participated in the supplemental data collection effort. RESULTS: An anti-smoking advertising campaign initiated by Vermont researchers was found to be the most cost effective in that it significantly reduced adolescent smoking prevalence at a low per capita cost. Next in order of cost effectiveness were California, Massachusetts, and Florida because behavioural outcomes were inconsistent across time and/or grades. California was ranked higher than the other two because it spent less per capita. Minnesota and Canada were ineffective at reducing adolescent smoking prevalence, and no comparison outcome data were available for Arizona. Four factors were found to be associated with increased cost effectiveness: (1) a greater use of message content that prior research suggests is efficacious with youth; (2) a more concentrated use of a single efficacious message; (3) an avoidance of unclear messages; and (4) an increased use of youthful spokespeople that adolescents could more readily identify with. No indication was found that depictions of smoking undermined campaign effectiveness by inadvertently implying that smoking was prevalent. CONCLUSIONS: The highly cost effective Vermont campaign can be used as a model for future efforts. It is estimated that 79% of the Vermont advertisements conveyed efficacious messages, 58% concentrated on a single efficacious message, 70% showed youthful spokespeople, and only 4% contained unclear messages. The results suggest that, in the less effective campaigns, as few as 25% of the advertisements contained messages that prior research indicates should be efficacious with youth, as few as 10% of the advertisements focused on one efficacious message, and up to 32% of the advertisements lacked clearcut messages. PMID- 10841589 TI - Spit (smokeless) tobacco use by high school baseball athletes in California. AB - OBJECTIVE: To describe the prevalence, patterns, and correlates of spit (smokeless) tobacco (ST) use in a sample of high school baseball athletes in California. DESIGN: This cross sectional study was a survey of 1226 baseball athletes attending 39 California high schools that were randomly selected from a list of all publicly supported high schools with baseball teams. At a baseball team meeting, athletes who agreed to participate and had parental consent completed the study questionnaire. To enhance the accuracy of self reported ST use status, a saliva sample was collected from each subject. The questionnaires and saliva samples were coded and salivary cotinine assay was performed on a random subsample of 5% of non-users who also were non-smokers. Biochemical assay indicated that 2% tested positive for cotinine inconsistent with self reported ST non-use. RESULTS: Overall, 46% had ever used ST and 15% were current users. Odds ratios and 95% confidence intervals (CI) suggested that, among high school baseball athletes, age, living in a rural area, being white, smoking cigarettes, drinking alcohol, not knowing about the adverse effects of ST, perceiving little personal risk associated with ST use, and believing that friends, role models, teammates, and same age baseball athletes in general used ST, increased the likelihood of being an ST user. CONCLUSION: The findings indicate that considerable experimentation with ST products occurs among high school baseball athletes in California, and many are current users. ST interventions targeting this population are needed to stop the transition from experimental ST use to tobacco dependence. Correlates of ST use for consideration in future intervention studies are identified. PMID- 10841590 TI - Reducing children's exposure to environmental tobacco smoke: the empirical evidence and directions for future research. AB - OBJECTIVE: To summarise the issues and empirical evidence for reduction of children's residential environmental tobacco smoke (ETS) exposure. DATA SOURCES: Literature was obtained by computer search, with emphasis on studies that included quantitative measures of ETS exposure in children's residences and interventions based on social learning theory. STUDY SELECTION: Review and empirical articles concerning ETS exposure were included and inferences were drawn based on a synthesis of these studies as contrasted with a quantitative meta-analysis. DATA SYNTHESIS: Interventions designed for residential/child ETS exposure control have included policy/legal regulations, minimal clinical services, and counselling services. Divorce court and adoption services have limited custody to protect children from ETS exposure. Controlled trials of clinicians' one time counselling services have shown null results. One controlled trial found that repeated physician ETS counselling increased parent cessation. Three trials found that repeated counselling/shaping procedures reduced quantitative estimates of ETS exposure in asthmatic children. CONCLUSIONS: Insufficient controlled studies of repeated session counselling procedures have been completed to determine efficacy for ETS exposure reduction, but evidence is promising. One time minimal interventions appear ineffective, but large scale studies may be warranted. No studies have been conducted to assess court or adoption agency regulations; no community ordinances for regulating residential ETS exposure have been invoked. Ethical and enforcement issues are discussed. PMID- 10841591 TI - A centralised telephone service for tobacco cessation: the California experience. AB - OBJECTIVE: To provide an overview of the California Smokers' Helpline, an increasingly popular telephone program for tobacco cessation in California since 1992. As many states, regions, and nations are contemplating various telephone programs as part of large scale anti-tobacco campaigns, this paper presents a practical model. DESIGN: The Helpline provides Californians with free cessation services that include counselling, self help quit kits, and cessation related information. Services are provided in six spoken languages plus a line for the hearing impaired. The program is promoted statewide by media campaigns, health care providers, local tobacco control programs, and the public school system. SETTING: The Helpline is centrally operated through the University of California, San Diego and provides services statewide via telephone. RESULTS: The Helpline has served over 100 000 tobacco users and has become the chief cessation resource for the Comprehensive Tobacco Control Program in California. Media was the most important referral source for Helpline callers (50%), followed by health care providers (20%). About one third of the callers were ethnic minorities and 17% were 24 years old or younger. Compared to California smokers in general, the callers were more dependent on nicotine and more likely to live with other smokers, but they were also more likely to have tried to quit recently and were more ready to try again. Two randomised trials have demonstrated the efficacy of the Helpline's counselling protocol. CONCLUSION: A centralised helpline operation can be an accessible and effective service for tobacco users and should be included in any large scale, comprehensive tobacco control program. PMID- 10841592 TI - Applying the quit & win contest model in the Vietnamese community in Santa Clara county. AB - OBJECTIVE: To evaluate the effectiveness of modifying and applying a Quit & Win contest model to Vietnamese Americans. DESIGN: Uncontrolled trial, multicomponent program, including two Quit & Win incentive contests, smoking cessation classes, videotape broadcasts, and newspaper articles. SUBJECTS AND SETTING: Vietnamese smokers living in Santa Clara County, California. MAIN OUTCOME MEASURES: Contest participation rates and quit rates at six month follow up; saliva cotinine validation of quitting. RESULTS: There were 57 eligible contest entrants to the 1995 contest, approximately 0.9% of the potential pool of smokers, and 32 entrants to the 1996 contest, approximately 0.5% of the potential pool. Overall, 48 of 49 (98%) individuals who said that they had quit smoking had validation of that fact by saliva cotinine testing. At six months, telephone follow up of 76 individuals revealed a self reported continued abstinence rate of 84.2%. CONCLUSION: Modification and application of the Quit & Win contest model for Vietnamese resulted not only in reasonable participation by Vietnamese male smokers, but also good success in initial quitting and an unexpectedly high abstinence rate at six month follow up. PMID- 10841593 TI - Cigarette consumption and sales of nicotine replacement products. AB - BACKGROUND: The first nicotine replacement product, Nicorette Gum, was marketed in 1984 as an adjuvant to help smokers quit smoking. In 1992, sales of nicotine patches were begun. Before 1996, nicotine gums and nicotine patches were prescribed by physicians and supplemented with behavioural counselling. Since 1996, nicotine gums and patches became available over the counter. OBJECTIVES: To examine the effect of sales of nicotine replacement products on national cigarette consumption. DESIGN: National time series quarterly cigarette consumption, sales of nicotine gums and patches data between 1976 and 1998 are used to estimate a time series autoregressive moving average intervention model. PARTICIPANTS: National reported statistics. MAIN OUTCOME MEASURES: Per capita cigarette sales. RESULTS: A 10% increase in sales of nicotine replacement products will lead to a 0.04% reduction in cigarette sales. The model indicates that a 0.076% reduction in cigarette consumption is associated with the availability of nicotine patches after 1992. The over the counter dummy variable (after 1996) has a negative sign, but is not significant, perhaps due to only a few quarters of data in the study period. CONCLUSIONS: Nicotine replacement products (nicotine gums and nicotine patches) play a significant role in reducing cigarette consumption, in addition to the negative effect of increasing cigarette price and the overall trend of declining cigarette consumption (as reflected by the time trend variable). The findings of this study suggest that additional efforts for promoting sales of nicotine replacement products will be another effective alternative to discouraging cigarette consumption. PMID- 10841594 TI - The evolution of miniaturized well plates. PMID- 10841595 TI - Test tube's end. PMID- 10841596 TI - Microbiological high throughput screening: an opportunity for the lead discovery process. AB - Microbial HTS has been implemented at Rhone-Poulenc Rorer through the development of a dedicated robotic platform. This robot (Turbo) has been designed with the aim of fully integrating microbial HTS into the lead discovery processes. Innovative solutions have been found to reach high throughput as well as flexibility. This opens up new prospects for solid-phase microbial screening, taking advantage of the easy implementation and the very low costs of such screens. The different types of microbial screens done in our laboratory, as well as the throughputs and outputs obtained, are described. Some of the specific aspects of microbial HTS, as compared to biochemical and cell-based assays, are also discussed. PMID- 10841597 TI - Identification of a high-affinity anti-phosphoserine antibody for the development of a homogeneous fluorescence polarization assay of protein kinase C. AB - In the last few years, fluorescence polarization (FP) has been applied to the development of robust, homogeneous, high throughput assays in molecular recognition research, such as ligand-protein interactions. Recently, this technology has been applied to the development of homogeneous tyrosine kinase assays, since there are high-affinity anti-phosphotyrosine antibodies available. Unlike tyrosine kinases, application of FP to assay development for serine/threonine kinases has been impeded because of lack of high-affinity anti phosphoserine/threonine antibodies. In the present study, we report the discovery of a high-affinity, monoclonal anti-phosphoserine antibody, 2B9, with a Kd of 250 +/- 34 pM for a phosphoserine-containing peptide tracer, fluorescein RFARKGS(PO(4))LRQKNV. Our data suggest that 2B9 is selective for fluorescein RFARKGS(PO(4))LRQKNV. The antibody and tracer have been used for the development of a competitive FP assay for protein kinase C (PKC) in 384-well plates. Phosphatidylserine, which enhances the kinase activity of PKC in a Ca(2+) dependent manner and has a structure similar to that of phosphoserine, did not interfere with binding of the peptide tracer to the antibody in the FP assay. The data indicate that the FP assay is more sensitive and robust than the scintillation proximity assay for PKC. The FP assay developed here can be used for rapid screening of hundreds of thousands of compounds for discovery of therapeutic leads for PKC-related diseases. PMID- 10841598 TI - An ultra-high throughput screening approach for an adenine transferase using fluorescence polarization. AB - We have developed a novel assay for measuring the activity of an enzyme that transfers multiple adenine-containing groups to an acceptor protein. The assay is based on fluorescence polarization (FP) technology in a 1536-well plate format. In the assay, a long wavelength fluorescence tracer, Texas Red (Rhodamine), was covalently conjugated to adenine of the donor substrate through a C(6) spacer arm. As a result of the transfer of the adenine-containing moieties to the acceptor protein substrate, the rotational correlation time of the Texas Red conjugate increased, hence increasing the degree of fluorescence polarization. The pharmacological profile and kinetics of the enzyme measured according to the FP method were consistent with those determined previously by conventional analysis. We have successfully executed a 250,000-compound high throughput screening program based on the FP assay method. The quality and validity of the assay were verified by a variety of statistical analyses. PMID- 10841599 TI - Implementing multilevel dynamic scheduling for a highly flexible 5-rail high throughput screening system. AB - As automation solves the bottleneck involved in drug screening, new bottlenecks present themselves. Some of these bottlenecks include sample management, hit picking and confirmation, and reagents lost as a result of incomplete runs. To keep up with the demands of a large HTS department, scientists spend a disproportionate amount of time simply feeding these systems with samples and reagents. Automating the sample management functions directly on the screening systems would solve this problem. With the use of online data analysis, an integrated sample store permits automated hit picking and confirmation. In addition to these issues, other bottlenecks are often caused by instrument malfunctions. A single lost run can now mean a loss of hundreds of plates and the reagents associated with their testing. A system was designed to include four assaying systems that are fed by an automated online sample repository system. Redundancy between the assaying systems allows for an extra level of error handling in case of a malfunction. The control of such a system requires a sophisticated scheduler/controller software package capable of coordinating the interaction between multiple systems and reacting to changes in the robotic environment in realtime. This paper discusses the design of the system as well as the requirements and selection of an appropriate scheduler/controller package. PMID- 10841600 TI - Determination of transition zone volume by transrectal ultrasound in patients with clinically benign prostatic hyperplasia: agreement with enucleated prostate adenoma weight. AB - PURPOSE: Accurately estimating transition zone volume is important for the medical or surgical management of symptomatic benign prostatic hyperplasia and determination of prostate specific antigen density of the transition zone. We evaluated whether preoperative transrectal ultrasound measurements of the transition zone predict enucleated adenoma weight. MATERIALS AND METHODS: We measured transition zone volume preoperatively using transrectal ultrasound and the prolate ellipsoid method in 50 patients with presumed benign prostatic hyperplasia who underwent suprapubic prostatectomy. Transition zone volume corresponds to the adenoma. Enucleated adenoma weight was then correlated with preoperatively determined transition zone volume. RESULTS: As measured by transrectal ultrasound, mean transition zone volume plus or minus standard deviation was 80.88 +/- 37.42 cc (range 31 to 200). Mean enucleated adenoma weight was 68.70 +/- 36.26 gm. (range 18 to 180). There was a statistically significant correlation of estimated transrectal ultrasound volume of the transition zone with enucleated prostate adenoma weight (r = 0.95, p <0.001). However, when prostate adenoma weight was determined using the formula, prostate adenoma weight = -6.00 + 0.92 x transition zone volume, we noted a significant difference in mean prostate adenoma weight and mean transition zone volume (p <0.001). Since the regression coefficient of transition zone volume was significantly different from 1, we identified no agreement of prostate adenoma weight with estimated transrectal ultrasound volume of the transition zone. CONCLUSIONS: These data reveal a significant difference in mean prostatic adenoma weight and mean transition zone volume. Although transition zone volume measurements are well described, clear agreements on such measurements should be obtained to determine transition zone volume more precisely. PMID- 10841601 TI - Paper alert. Immunology. PMID- 10841602 TI - Paper alert. Immunology. PMID- 10841603 TI - Web alert. Lymphocyte development. Immunological techniques. PMID- 10841604 TI - Preoperative checking of IMA(s): to do or not to do? PMID- 10841605 TI - Retrograde perfusion of oxygenated blood during off pump revascularization to avoid ischemia. PMID- 10841606 TI - Long-term survival after bronchial sleeve resection in relation to nodal involvement. PMID- 10841607 TI - Re: Vegetarianism and dental fluorosis among children in a high fluoride area of northern Tanzania (1999;9: 3-11) PMID- 10841608 TI - Funding adolescent medicine fellowship training. PMID- 10841609 TI - Completion of hepatitis B vaccination series in school-based health centers. PMID- 10841610 TI - Photo quiz. Echinococcal cyst of the fifth thoracic vertebra in an otherwise healthy young woman. PMID- 10841611 TI - Special issue dedicated to the memory of Therese M. Cotton. PMID- 10841612 TI - Normal T, B, and NK cell counts in healthy donors at 1 year after blood stem cell harvesting. PMID- 10841613 TI - Donor lymphocyte infusions for CML: possible effects of age and mobilization. PMID- 10841614 TI - Benzene and multiple myeloma: appraisal of the scientific evidence. PMID- 10841616 TI - Paper alert. Neurobiology. PMID- 10841615 TI - DAP-kinase CpG island methylation in acute myeloid leukemia: methodology versus biology? PMID- 10841617 TI - Neuromuscular disorders: gene location. PMID- 10841618 TI - Mitochondrial encephalomyopathies: gene mutation. PMID- 10841619 TI - Historical studies in neuromuscular disorders. PMID- 10841620 TI - A modified alignment of human and rodent 5' untranslated sequences of the acetylcholine receptor epsilon subunit gene reveals additional regions of high homology. PMID- 10841621 TI - Underwater and Hyperbaric Medicine, abstracts from the literature. PMID- 10841623 TI - [3d National Congress on Transfusion Safety and Hemovigilance. Lille, France, 16 7 September 1999. Proceedings and abstracts]. PMID- 10841624 TI - UK speculated to permit stem cell research. PMID- 10841625 TI - RE: M Thomas. The change of cost: reference-based pricing and the statins. 1999;15:535-8. PMID- 10841626 TI - Depomedroxyprogesterone acetate use and weight gain among adolescents. PMID- 10841627 TI - The normal range and determinants of the intrinsic heart rate in man. PMID- 10841628 TI - Gestational diabetes in primary care. PMID- 10841629 TI - Why big lies matter: lessons from the Bezwoda affair. PMID- 10841631 TI - Stem cells show their potential. PMID- 10841632 TI - Clarity on the diagnosis line. PMID- 10841633 TI - [Evaluation of the knowledge and attitudes of dental personnel about acquired immunodeficiency syndrome (AIDS)]. PMID- 10841634 TI - The process of forming solid compacts by the application of pressure to a powder bed is not the preserve of pharmacy. PMID- 10841635 TI - J-shaped relation between blood pressure and stroke. PMID- 10841636 TI - 'Infectious web'. AB - This issue of 'Infectious Web' includes web-sites related to AIDS/HIV, pathogenic characteristics and resistance to Staphylococcus spp., diagnostic and clinical aspects of arthritis, and comprehensive information resources on malaria, cystic fibrosis and biological weapons. PMID- 10841637 TI - Synthesis of trans-2-(1-aryl-1-methylethyl)cyclohexylamines. AB - As a first example of opening a secondary aziridine with a tertiary carbanion, the title amines (3a-c, aryl = phenyl, 4-tert-butylphenyl,2-naphthyl) were synthesized by opening N-(diphenylphosphinoyl)-7-azabicyclo[4.1.0]heptane, aziridine 1, with the corresponding alpha-potassium isopropylarenes, followed by a hydrolysis of the resulting phosphinamides 2a-c. PMID- 10841638 TI - Proceedings of the Multimodality Approach to Lung Cancer Symposium. Vail , Colorado, USA. February 1998. PMID- 10841639 TI - CagA protein from Helicobacter pylori is a Trojan Horse to epithelial cells. PMID- 10841640 TI - Reduced access to gastric motility drug will not sit well with many physicians or patients. PMID- 10841641 TI - Telomere deficiency worsens liver disease. PMID- 10841642 TI - Image of the month. Recurrent painless, massive gastrointestinal (GI) bleeding obscure in origin. Dieulafoy's lesion. PMID- 10841643 TI - Dukes of the Dukes classification. PMID- 10841644 TI - Insulin resistance influence iron metabolism and hepatic steatosis in type II diabetes. PMID- 10841645 TI - Short-segment Barrett's: a significant lesion. PMID- 10841646 TI - A "G insertion" at nucleic acids 481 of the H. pylori fldA gene is not associated with gastric MALT lymphoma. PMID- 10841647 TI - Adenocarcinoma and non-Hodgkin's lymphoma involving the prostate. PMID- 10841648 TI - Inflammatory pseudotumour of the lymph node in an HIV-positive individual. PMID- 10841649 TI - Microglandular adenosis arising in chronic paranasal sinusitis. PMID- 10841650 TI - Sclerosing neurofibroma. PMID- 10841651 TI - Well-differentiated lipoma-like liposarcoma of the caecum. PMID- 10841652 TI - Multicentric clear cell adenocarcinoma in the urinary bladder and the urethral diverticulum: evidence of origin of clear cell adenocarcinoma of the female lower urinary tract from Mullerian duct remnants. PMID- 10841653 TI - Adenomatous hyperplasia of the rete ovarii. PMID- 10841654 TI - [Injuries caused by traffic accidents in Spain: a call to action]. PMID- 10841655 TI - [Some achievements and challenges]. PMID- 10841656 TI - [Use of drugs and HIV. A review of recent history as a foundation of new prevention policies]. PMID- 10841657 TI - [The role of primary care in an integrated organization of services and its contribution to global efficiency]. PMID- 10841658 TI - Angina--an Indian disease. PMID- 10841659 TI - Pictorial CME. True precocious puberty secondary to hydrocephalus. PMID- 10841660 TI - Novel antipsychotic use in schizophrenia. PMID- 10841662 TI - Proceedings of the 1997 Conference on Childhood Bipolarity. Bethesda, Maryland, USA. PMID- 10841661 TI - Hla-DR2-restricted responses to proteolipid protein 95-116 peptide cause autoimmune encephalitis in transgenic mice. AB - In multiple sclerosis (MS) patients who carry the Class II major histocompatibility (MHC) type HLA-DR2, T cells specific for amino acids 95-116 in the proteolipid protein (PLP) are activated and clonally expanded. However, it remains unclear whether these autoreactive T cells play a pathogenic role or, rather, protect against the central nervous system (CNS) damage. We have addressed this issue, using mice transgenic for the human MHC class II region carrying the HLA-DR2 (DRB1* 1502) haplotype. After stimulating cultured lymph node cells repeatedly with PLP95-116, we generated 2 HLA-DR2-restricted, PLP95 116-specific T-cell lines (TCLs) from the transgenic mice immunized with this portion of PLP. The TCLs were CD4+ and produced T-helper 1 (Th1) cytokines in response to the peptide. These TCLs were adoptively transferred into RAG-2/2 mice expressing HLA-DR2 (DRG1* 1502) molecules. Mice receiving 1 of the TCLs developed a neurological disorder manifested ataxic movement without apparent paresis on day 3, 4, or 5 after cell transfer. Histological examination revealed inflammatory foci primarily restricted to the cerebrum and cerebellum, in association with scattered demyelinating lesions in the deep cerebral cortex. These results support a pathogenic role for PLP95-116-specific T cells in HLA DR2+ MS patients, and shed light on the possible correlation between autoimmune target epitope and disease phenotype in human CNS autoimmune diseases. PMID- 10841664 TI - 22nd Annual Carrell-Krusen Symposium. Dallas, Texas, USA. February 24-25, 2000. Abstracts. PMID- 10841663 TI - Effects of ascorbate supplements on urinary oxalate and risk of kidney stones. PMID- 10841666 TI - ICANL and ACR nuclear medicine accreditation: a comparison. PMID- 10841665 TI - On mortality from ischemic heart disease in women with very late menopause. PMID- 10841667 TI - Authorship: rite, right, or write of passage? PMID- 10841668 TI - Smoking cessation in primary care clinics. PMID- 10841669 TI - Preventative care in chronic liver diseases. PMID- 10841670 TI - Round trip radical reactions from acyclic precursors to tricyclo [5.3.1.0]undecanes. A new cascade radical cyclization approach to (+/-) isogymnomitrene and (+/-)-gymnomitrene. AB - The synthesis and "round trip radical cyclization" of 11-iodo-2,7,11 trimethyldodec-6-en-5-one are described. The round trip cyclization is a sequence of 5-exo, 6-endo, and 5-exo cyclizations in which the last radical cyclization occurs at the same carbon atom as the initial radical generation. The key second (6-endo) cyclization produces two stereoisomers, one of which cyclizes efficiently to isogymnomitrene ketone, while the other cyclizes inefficiently to gymnomitrene ketone. Efforts to influence the kinetic or thermodynamic outcome of the second cyclization were not successful, and the results are contrasted with a related cyclization of Jung and Rayle where thermodynamic control was readily established. PMID- 10841671 TI - Direct transacylation of 2,2,2-trihaloethyl esters with amines and alcohols using phosphorus (III) reagents for reductive fragmentation and in situ activation. AB - Amides and esters have been synthesized from 2,2,2-trihaloethyl esters in one pot using phosphorus-(III) reagents as reductants, with resultant carboxylate activation as an acyloxyphosphonium intermediate, and in situ trapping by amine or alcohol nucleophiles. Secondary and tertiary amides were synthesized, including a dipeptide, in good yields using hexamethylphosphorous triamide (Me2N)3P, as reducing agent. Optimal yields of esters derived from primary and secondary alcohols were obtained using tributylphosphine and DMAP. Tribromoethyl esters provided yields superior to those obtained with trichloroethyl esters. PMID- 10841672 TI - [Proceedings of the 1st Pediatric Congress of Bosnia and Herzegovina. Sarajevo, October 15-17, 1998]. PMID- 10841674 TI - [Homeopathy]. PMID- 10841673 TI - [Be aware of insidious changes in practice]. PMID- 10841675 TI - [Homeopathy]. PMID- 10841676 TI - Posthypoxic myoclonus. Workshop proceedings. Washington DC, USA. March 21-22, 1999. PMID- 10841677 TI - Microsatellite instability in colorectal cancer. PMID- 10841678 TI - Microsatellite instability in colorectal cancer. PMID- 10841679 TI - Urgent colonoscopy for the diagnosis and treatment of severe diverticular hemorrhage. PMID- 10841680 TI - Urgent colonoscopy for the diagnosis and treatment of severe diverticular hemorrhage. PMID- 10841681 TI - Urgent colonoscopy for the diagnosis and treatment of severe diverticular hemorrhage. PMID- 10841682 TI - Urgent colonoscopy for the diagnosis and treatment of severe diverticular hemorrhage. PMID- 10841683 TI - Urgent colonoscopy for the diagnosis and treatment of severe diverticular hemorrhage. PMID- 10841684 TI - Urgent colonoscopy for the diagnosis and treatment of severe diverticular hemorrhage. PMID- 10841685 TI - Regional enteritis associated with enterovirus in a patient with X-linked agammaglobulinemia. PMID- 10841686 TI - Prediction of adverse outcomes in children with sickle cell disease. PMID- 10841687 TI - Supplemental perioperative oxygen to reduce surgical-wound infections. PMID- 10841688 TI - Supplemental perioperative oxygen to reduce surgical-wound infections. PMID- 10841689 TI - Pyomyositis complicating acute bacterial endocarditis in an intravenous drug user. PMID- 10841690 TI - Unesco backs 'science for debt' plan. PMID- 10841691 TI - German research agency 'doesn't stifle creativity', say 1,600 scientists. PMID- 10841692 TI - Shareholder sues Celera over loss. PMID- 10841693 TI - Jane Gray (1929-2000). PMID- 10841694 TI - European public remains sceptical. PMID- 10841695 TI - The morcellator knife: a new laparoscopic instrument for supracervical hysterectomy and morcellation. AB - BACKGROUND: Cutting the cervix, morcellation, and extraction of the uterus and myomata remain major problems in endoscopic surgery. We developed an efficient, safe, reusable, and inexpensive instrument to cut the cervix and morcellate the uterus and myomata: the morcellator knife. INSTRUMENT: The morcellator knife is a classic lancet with an interchangeable blade, transformed into an endoscopic instrument that can be inserted easily through a 10-mm-diameter trocar. The blade has an automatic retraction system and is set in the standby position, ensuring security. Cutting the cervix and uterine or myoma fragmentation are easy. The mass to be cut is held between two grasping forceps for easy cutting with the blade, under permanent visual control. After morcellation, extraction of the masses is performed through a posterior culdotomy. EXPERIENCE: We have used this morcellator knife in 54 subtotal hysterectomies and 16 myomectomies. There were no complications during the procedures. Morcellation lasted 3-14 minutes and involved the use of an average of two to three blades. CONCLUSION: The morcellator knife is a simple, safe, reusable, and inexpensive instrument with a low maintenance cost. PMID- 10841696 TI - Comparison of oral misoprostol and oxytocin for labor induction. PMID- 10841697 TI - What is the earliest non-invasive malignant lesion of the larynx? PMID- 10841698 TI - [10th International Congress on Anti-Cancer Treatment Paris, Palais de Congres, 31 January - 3 February 2000. On the subject of new treatments in oncology]. PMID- 10841699 TI - Factors in risk perception. PMID- 10841700 TI - [XXIst Days on Medical Biophysics. Horni Porici, May 27-29, 1998. Proceedings]. PMID- 10841701 TI - Gifts to physicians from industry. PMID- 10841702 TI - Space biology. Goldin shakes up NASA's life sciences program. PMID- 10841703 TI - Human Genome Project. Chromosome 21 done, phase two begun. PMID- 10841704 TI - Structural genomics. Protein data justice for all. PMID- 10841705 TI - Global warming. Some coral bouncing back from El Nino. PMID- 10841706 TI - Climate change. Panel estimates possible carbon 'sinks'. PMID- 10841707 TI - Circadian rhythms. Two feedback loops run mammalian clock. PMID- 10841708 TI - Human genome. Mapping a subtext in our genetic book. PMID- 10841709 TI - Paleoanthropology. A glimpse of humans' first journey out of Africa. PMID- 10841710 TI - Gene therapy on trial. PMID- 10841711 TI - Improving gene therapy's tool kit. PMID- 10841712 TI - Gene therapy's web of corporate connections. PMID- 10841713 TI - Rice, races, and riches. PMID- 10841714 TI - Dialog on depression. PMID- 10841715 TI - Dialog on depression. PMID- 10841716 TI - Induced damage in the developing brain. PMID- 10841717 TI - Induced damage in the developing brain. PMID- 10841718 TI - Risk management. Science and the precautionary principle. PMID- 10841719 TI - Immunology. A touch of antibody class. PMID- 10841720 TI - Black-footed ferret recovery. PMID- 10841721 TI - Diversity of human alpha beta T cell receptors. PMID- 10841722 TI - Human Genome Project. And the gene number is...? PMID- 10841723 TI - Conservation biology. Orangutans face extinction in the wild. PMID- 10841724 TI - European science. Spain opens coffers to keep talent at home. PMID- 10841725 TI - Conservation biology. California team to map rare species' DNA. PMID- 10841726 TI - DNA computing. Hairpins trigger an automatic solution. PMID- 10841727 TI - Japan. Exposure levels tracked around nuclear accident. PMID- 10841728 TI - Journal publishing. Harvard researcher named NEJM editor. PMID- 10841729 TI - 2001 budget. NIH headed for big boost, others struggle. PMID- 10841730 TI - Linguistics. Learning the world's languages--before they vanish. PMID- 10841731 TI - Zebrafish earns its stripes in genetic screens. PMID- 10841732 TI - Taiwan. Researchers say new institutes offer them a chance 'to do good science'. PMID- 10841734 TI - A home for Bambiraptor. PMID- 10841733 TI - HIV/AIDS: the peril of pseudoscience. PMID- 10841735 TI - Intellectual property rights. PMID- 10841736 TI - Intellectual property rights. PMID- 10841737 TI - Is there an intrinsic period of the circadian clock? PMID- 10841738 TI - Evolutionary genetics. Sinless originals. PMID- 10841739 TI - Microbiology. When being hyper keeps you fit. PMID- 10841741 TI - Epistola chirurgica. VI. Counseling and consultation. PMID- 10841740 TI - Robotic vision. Neuromorphic vision sensors. PMID- 10841742 TI - [CA-125 as response parameter in ovarian carcinoma]. PMID- 10841743 TI - [Cerebral hemorrhage after diving]. PMID- 10841744 TI - [Feeding of critically ill patients]. PMID- 10841745 TI - [Reference value for plasma progesterone content]. PMID- 10841747 TI - 25th International Symposium on Ultrasonic Imaging and Tissue Characterization. Arlington, Virginia, USA. May 22-24, 2000. Abstracts. PMID- 10841746 TI - [Dr. Jeannette (Nettie) Donker-Voet, first female veterinarian in the Netherlands]. PMID- 10841748 TI - [Special issue on rheumatic diseases, dedicated to Professor Dr. Gert Klein, on the occasion of his 60th birthday]. PMID- 10841749 TI - [New trends in vascular surgery: their influence on structures and organization. 9th Workshop: Good Clinical Practice. Melsungen, November 27-28, 1998. Abstracts]. PMID- 10841750 TI - The 9th Annual Congress of The Japan Bi-Digital O-Ring Test Medical Society. Tokyo, Japan, July 24-25, 1999. Abstracts. PMID- 10841751 TI - The 15th Annual International Symposium on Acupuncture and Electro-Therapeutics. New York City, New York, USA. October 21-24, 1999. Abstracts. PMID- 10841752 TI - International standards for the assessment of the risk of thermal strain on clothed workers in hot environments: comment. PMID- 10841753 TI - Exploring the calcium-binding site in photosystem II membranes by solid-state (113)Cd NMR. AB - Calcium (Ca(2+)) is an essential cofactor for photosynthetic oxygen evolution. Although the involvement of Ca(2+) at the oxidizing side of photosystem II of plants has been known for a long time, its ligand interactions and mode of action have remained unclear. In the study presented here, (113)Cd magic-angle spinning solid-state NMR spectroscopy is used to probe the Ca(2+)-binding site in the water-oxidizing complex of (113)Cd(2+)-substituted PS2. A single NMR signal 142 ppm downfield from Cd(ClO(4))(2).2H(2)O was recorded from Cd(2+) present at the Ca(2+)-binding site. The anisotropy of the signal is small, as indicated by the absence of spinning side bands. The signal intensity is at its maximum at a temperature of -60 degrees C. The line width of the proton signal in a WISE (wide line separation) two-dimensional (1)H-(113)Cd NMR experiment demonstrates that the signal arises from Cd(2+) in a solid and magnetically undisturbed environment. The chemical shift, the small anisotropy, and the narrow line of the (113)Cd NMR signal provide convincing evidence for a 6-fold coordination, which is achieved partially by oxygen and partially by nitrogen or chlorine atoms in otherwise a symmetric octahedral environment. The absence of a (113)Cd signal below -70 degrees C suggests that the Ca(2+)-binding site is close enough to the tetramanganese cluster to be affected by its electron spin state. To our knowledge, this is the first report for the application of solid-state NMR in the study of the membrane-bound PS2 protein complex. PMID- 10841754 TI - Tracing the D-pathway in reconstituted site-directed mutants of cytochrome c oxidase from Paracoccus denitrificans. AB - Heme-copper terminal oxidases use the free energy of oxygen reduction to establish a transmembrane proton gradient. While the molecular mechanism of coupling electron transfer to proton pumping is still under debate, recent structure determinations and mutagenesis studies have provided evidence for two pathways for protons within subunit I of this class of enzymes. Here, we probe the D-pathway by mutagenesis of the cytochrome c oxidase of the bacterium Paracoccus denitrificans; amino acid replacements were selected with the rationale of interfering with the hydrophilic lining of the pathway, in particular its assumed chain of water molecules. Proton pumping was assayed in the reconstituted vesicle system by a stopped-flow spectroscopic approach, allowing a reliable assessment of proton translocation efficiency even at low turnover rates. Several mutations at positions above the cytoplasmic pathway entrance (Asn 131, Asn 199) and at the periplasmic exit region (Asp 399) led to complete inhibition of proton pumping; one of these mutants, N131D, exhibited an ideal decoupled phenotype, with a turnover comparable to that of the wild-type enzyme. Since sets of mutations in other positions along the presumed course of the pathway showed normal proton translocation stoichiometries, we conclude that the D-pathway is too wide in most areas above positions 131/199 to be disturbed by single amino acid replacements. PMID- 10841755 TI - Proton equilibria in the manganese cluster of photosystem II control the intensities of the S(0) and S(2) state g approximately 2 electron paramagnetic resonance signals. AB - We have studied the pH effect on the S(0) and S(2) multiline electron paramagnetic resonance (EPR) signals from the water-oxidizing complex of photosystem II. Around pH 6, the maximum signal intensities were detected. On both the acidic and alkaline sides of pH 6, the intensities of the EPR signals decreased. Two pKs were determined for the S(0) multiline signal; pK(1) = 4.2 +/- 0.2 and pK(2) = 8.0 +/- 0.1, and for the S(2) multiline signal the pKs were pK(1) = 4.5 +/- 0.1 and pK(2) = 7.6 +/- 0.1. The intensity of the S(0)-state EPR signal was partly restored when the pH was changed from acidic or alkaline pH back to pH approximately 6. In the S(2) state we observed partial recovery of the multiline signal when going from alkaline pH back to pH approximately 6, whereas no significant recovery of the S(2) multiline signal was observed when the pH was changed from acidic pH back to pH approximately 6. Several possible explanations for the intensity changes as a function of pH are discussed. Some are ruled out, such as disintegration of the Mn cluster or decay of the S states and formal Cl( ) and Ca(2+) depletion. The altered EPR signal intensities probably reflect the protonation/deprotonation of ligands to the Mn cluster or the oxo bridges between the Mn ions. Also, the possibility of decreased multiline signal intensities at alkaline pH as an effect of changed redox potential of Y(Z) is put forward. PMID- 10841756 TI - Role of the glutamate 332 residue in the transglycosylation activity of ThermusMaltogenic amylase. AB - A sequence alignment shows that residue 332 is conserved as glutamate in maltogenic amylases (MAases) and in other related enzymes such as cyclodextrinase and neopullulanase, while the corresponding position is conserved as histidine in alpha-amylases. We analyzed the role of Glu332 in the hydrolysis and the transglycosylation activity of Thermus MAase (ThMA) by site-directed mutagenesis. Replacing Glu332 with histidine reduced transglycosylation activity significantly, but enhanced hydrolysis activity on alpha-(1,3)-, alpha-(1,4)-, and alpha-(1,6)-glycosidic bonds relative to the wild-type (WT) enzyme. The mutant Glu332Asp had catalytic properties similar to those of the WT enzyme, but the mutant Glu332Gln resulted in significantly decreased transglycosylation activity. These results suggest that an acidic side chain at position 332 of MAase plays an important role in the formation and accumulation of transfer products by modulating the relative rates of hydrolysis and transglycosylation. From the structure, we propose that an acidic side chain at position 332, which is located in a pocket, is involved in aligning the acceptor molecule to compete with water molecules in the nucleophilic attack of the glycosyl-enzyme intermediate. PMID- 10841757 TI - Crystal structures of Giardia lamblia guanine phosphoribosyltransferase at 1.75 A(,). AB - Giardia lamblia, the protozoan parasite responsible for giardiasis, requires purine salvage from its host for RNA and DNA synthesis. G. lamblia expresses an unusual purine phosphoribosyltransferase with a high specificity for guanine (GPRTase). The enzyme's sequence significantly diverges from those of related enzymes in other organisms. The transition state analogue immucillinGP is a powerful inhibitor of HGXPRTase from malaria [Li, C. M., et al. (1999) Nat. Struct. Biol. 6, 582-587] and is also a 10 nM inhibitor of G. lamblia GPRTase. Cocrystallization of GPRTase with immucillinGP led unexpectedly to a GPRTase.immucillinG binary complex with an open catalytic site loop. Diffusion of ligands into preformed crystals gave a GPRTase.immucillinGP.Mg(2+).pyrophosphate complex in which the open loop is stabilized by crystal contacts. G. lamblia GPRTase exhibits substantial structural differences from known purine phosphoribosyltransferases at positions remote from the catalytic site, but conserves most contacts to the bound inhibitor. The filled catalytic site with an open catalytic loop provides insight into ligand binding. One active site Mg(2+) ion is chelated to pyrophosphate, but the other is chelated to two conserved catalytic site carboxylates, suggesting a role for these amino acids. This arrangement of Mg(2+) and pyrophosphate has not been reported in purine phosphoribosyltransferases. ImmucillinG in the binary complex is anchored by its 9-deazaguanine group, and the iminoribitol is disordered. No Mg(2+) or pyrophosphate is detected; thus, the 5'-phosphoryl group is needed to immobilize the iminoribitol prior to magnesium pyrophosphate binding. Filling the catalytic site involves (1) binding the purine ring, (2) anchoring the 5'-phosphate to fix the ribosyl group, (3) binding the first Mg(2+) to Asp125 and Glu126 carboxyl groups and binding Mg(2+).pyrophosphate, and (4) closing the catalytic site loop and formation of bound (Mg(2+))(2). pyrophosphate prior to catalysis. Guanine specificity is provided by two peptide carbonyl oxygens hydrogen-bonded to the exocyclic amino group and a weak interaction to O6. Transition state formation involves N7 protonation by Asp129 acting as the general acid. PMID- 10841758 TI - Evidence for unfolding of the single-stranded GCCA 3'-End of a tRNA on its aminoacyl-tRNA synthetase from a stacked helical to a foldback conformation. AB - The conformation of a tRNA in its initial contact with its cognate aminoacyl-tRNA synthetase was investigated with the Escherichia coli glutamyl-tRNA synthetase tRNA(Glu) complex. Covalent complexes between the periodate-oxidized tRNA(Glu) and its synthetase were obtained. These complexes are specific since none were formed with any other oxidized E. coli tRNA. The three major residues cross linked to the 3'-terminal adenosine of oxidized tRNA(Glu) are Lys115, Arg209, and Arg48. Modeling of the tRNA(Glu)-glutamyl-tRNA synthetase based on the known crystal structures of Thermus thermophilus GluRS and of the E. coli tRNA(Gln) glutaminyl-tRNA synthetase complex shows that these three residues are located in the pocket that binds the acceptor stem, and that Lys115, located in a 26 residue loop closed by coordination to a zinc atom in the tRNA acceptor stem-binding domain, is the first contact point of the 3'-terminal adenosine of tRNA(Glu). In our model, we assume that the 3'-terminal GCCA single-stranded segment of tRNA(Glu) is helical and extends the stacking of the acceptor stem. This assumption is supported by the fact that the 3' CCA sequence of tRNA(Glu) is not readily circularized in the presence of T4 RNA ligase under conditions where several other tRNAs are circularized. The two other cross-linked sites are interpreted as the contact sites of the 3'-terminal ribose on the enzyme during the unfolding and movement of the 3'-terminal GCCA segment to position the acceptor ribose in the catalytic site for aminoacylation. PMID- 10841759 TI - The ATT strand of AAT.ATT trinucleotide repeats adopts stable hairpin structures induced by minor groove binding ligands. AB - AAT.ATT is the most abundant and also the most frequently polymorphic class of trinucleotide repeats in the human genome. To characterize its structural properties and conformational changes induced by minor groove ligands, (AAT)(6) and (ATT)(6) oligomers as well as their complexes with DAPI were investigated by electrophoretic mobility and UV thermal stability as well as fluorescence and NMR spectroscopy. The results show that individual (AAT)(6) and (ATT)(6) strands exist principally as monomeric non-hydrogen-bonded structures. Their individual interaction with DAPI induces the formation of base-paired structures with different thermal stabilities by quite spectroscopically distinct binding mechanisms. In the presence of DAPI, (ATT)(6) forms a monomeric hairpin structure stabilized by two ligands located in the minor groove with a strong apparent binding constant of 3.4 x 10(6) M(-)(1). The DAPI-induced (ATT)(6) hairpin is characterized by well-stacked A.T Watson-Crick and T.T wobble base pairs, a high electrophoretic mobility, and a melting temperature of 41 degrees C. Interaction of DAPI with the complementary (AAT)(6) strand favors less stable base-paired structures, and the results are consistent with electrostatic and hydrogen-bond interactions of the ligand with the phosphodiester backbone of (AAT)(6) by minor involvement of DNA bases. PMID- 10841760 TI - Conserved phosphoprotein interaction motif is functionally interchangeable between ataxin-7 and arrestins. AB - Olivopontocerebellar atrophy with retinal degeneration is a hereditary neurodegenerative disorder that belongs to the subtype II of the autosomal dominant cerebellar ataxias and is characterized by early-onset cerebellar and macular degeneration preceded by diagnostically useful tritan colorblindness. The gene mutated in the disease (SCA7) has been mapped to chromosome 3p12-13.5, and positional cloning identified the cause of the disease as CAG repeat expansion in this gene. The SCA7 gene product, ataxin-7, is an 897 amino acid protein with an expandable polyglutamine tract close to its N-terminus. No clues to ataxin-7 function have been obtained from sequence database searches. Here we report that ataxin-7 has a motif of ca. 50 amino acids, related to the phosphate-binding site of arrestins. To test the relevance of this sequence similarity, we introduced the putative ataxin-7 phosphate-binding site into visual arrestin and beta arrestin. Both chimeric arrestins retain receptor-binding affinity and show characteristic high selectivity for phosphorylated activated forms of rhodopsin and beta-adrenergic receptor, respectively. Although the insertion of a Gly residue (absent in arrestins but present in the putative phosphate-binding site of ataxin-7) disrupts the function of visual arrestin-ataxin-7 chimera, it enhances the function of beta-arrestin-ataxin-7 chimera. Taken together, our data suggest that the arrestin-like site in the ataxin-7 sequence is a functional phosphate-binding site. The presence of the phosphate-binding site in ataxin-7 suggests that this protein may be involved in phosphorylation-dependent binding to its protein partner(s) in the cell. PMID- 10841761 TI - Determination of pK(a) values of carboxyl groups in the N-terminal domain of rat CD2: anomalous pK(a) of a glutamate on the ligand-binding surface. AB - The ligand-binding surface of the T-lymphocyte glycoprotein CD2 has an unusually high proportion of charged residues, and ionic interactions are thought to play a significant role in defining the ligand specificity and binding affinity of CD2 with the structurally homologous ligands CD48 (in rodents) and CD58 (in humans). The determination of the electrostatic properties of these proteins can therefore contribute to our understanding of structure-activity relationships for these adhesion complexes that underpin T-cell adhesion to antigen-presenting cells. In this study, we investigated the pH titration behavior of the carboxyl groups of the N-terminal domain of rat CD2 (CD2d1) using the chemical shifts of backbone amide nitrogen-15 ((15)N) and proton NMR resonances, and carboxyl carbon-13 ((13)C) signals. The analysis revealed the presence of a glutamate (Glu41) on the binding surface of rat CD2 with an unusually elevated acidity constant (pK(a) = 6.73) for CD2d1 samples at 1.2 mM concentration. pH titration of CD2d1 at low protein concentration (0.1 mM) resulted in a slight decrease of the measured pK(a) of Glu41 to 6.36. The ionization of Glu41 exhibited reciprocal interactions with a second glutamate (Glu29) in a neighboring location, with both residues demonstrating characteristic biphasic titration behavior of the carboxyl (13)C resonances. Measurements at pH 5.5 of the two-bond deuterium isotope shift for the (13)C carboxyl resonances for Glu41 and Glu29 [(2)DeltaC(delta)(O(epsilon)D) = 0.2 and 0.1 ppm, respectively] were consistent with the assignment of the anomalous pK(a) to Glu41, under the strong influence of Glu29. The characterization of single site mutations of CD2d1 residues Glu41 and Glu29 to glutamine confirmed the anomalous pK(a) for Glu41, and indicated that electrostatic interaction with the Glu29 side chain is a significant contributing influence for this behavior in the wild-type protein. The implications of these observations are discussed with respect to recent structural and functional analyses of the interaction of rat CD2 with CD48. In particular, CD2 Glu41 must be a candidate residue to explain the previously reported strong pH dependence of binding of these two proteins in vitro. PMID- 10841762 TI - Toward a high-resolution structure of phospholamban: design of soluble transmembrane domain mutants. AB - Determination of a high-resolution structure of the phospholamban (PLB) transmembrane domain by X-ray crystallography or NMR is handicapped by the hydrophobic nature of the peptide. Interestingly, the crystal structure of the five-stranded parallel coiled-coil oligomerization domain from cartilage oligomeric matrix protein (COMPcc) shows marked similarities to a model proposed for the pentameric transmembrane domain of PLB. Contrary to the putative coiled coil domain of PLB, COMPcc contains mostly hydrophilic amino acids on the surface, resulting in a soluble molecule. Here, we report the design of soluble PLB transmembrane domain variants by combining the surface residues of COMPcc and the hydrophobic interior of the transmembrane domain of PLB. The soluble PLB variants formed pentameric structures as revealed by analytical ultracentrifugation. After redox shuffling, they showed unspecific disulfide bridge patterns similar to that of the chemically synthesized wild-type PLB transmembrane domain. These results suggest a structural homology between the soluble PLB mutants and the wild-type PLB transmembrane domain. Together with the data reported in the literature, they furthermore indicate that residues Leu37, Ile40, Leu44, and Ile47 of the PLB sequence specify pentamer formation. In contrast, a designed recombinant COMPcc mutant, COMP-ARCC, which was engineered to contain the two PLB cysteines that potentially could form an interchain disulfide bridge, formed a specific disulfide bond pattern. This finding indicates structural differences between the transmembrane domain of PLB and COMPcc. The soluble PLB variants may be used to determine a high-resolution structure of the PLB pentamer by X-ray crystallography. PMID- 10841763 TI - Novel insights into catalytic mechanism from a crystal structure of human topoisomerase I in complex with DNA. AB - Human topoisomerase I helps to control the level of DNA supercoiling in cells and is vital for numerous DNA metabolic events, including replication, transcription, and recombination. The 2.6 A crystal structure of human topoisomerase I in noncovalent complex with a DNA duplex containing a cytosine at the -1 position of the scissile strand rather than the favored thymine is reported. The hydrogen bond between the O2 position of this -1 base and the epsilon-amino of the conserved Lys-532 residue, the only base-specific contact observed previously in the human topoisomerase I-DNA interaction, is maintained in this complex. Several unique features of this structure, however, have implications for the DNA-binding and active-site mechanisms of the enzyme. First, the ends of the DNA duplex were observed to shift by up to 5.4 A perpendicular to the DNA helical axis relative to structures reported previously, suggesting a novel degree of plasticity in the interaction between human topoisomerase I and its DNA substrate. Second, 12 additional residues at the NH(2) terminus of the protein (Trp-203-Gly-214) could be built in this structure, and they were found to pack against the putative hinge region implicated in the clamping of the enzyme around duplex DNA. Third, a water molecule was observed adjacent to the scissile phosphate and the active site residues; the potential specific base character of this solvent molecule in the active-site mechanism of the enzyme is discussed. Fourth, the scissile phosphate group was found to be rotated by 75 degrees, bringing Lys-532 into hydrogen-bonding distance of one of the nonbridging phosphate oxygens. This orientation of the scissile phosphate group implicates Lys-532 as a fifth active site residue, and also mimics the orientation observed for the 3'-phosphotyrosine linkage in the covalent human topoisomerase I-DNA complex structure. The implications of these structural features for the mechanism of the enzyme are discussed, including the potential requirement for a rotation of the scissile phosphate group during DNA strand cleavage and covalent attachment. PMID- 10841764 TI - A bacterioferritin from the strict anaerobe Desulfovibrio desulfuricans ATCC 27774. AB - A bacterioferritin was isolated from the anaerobic bacterium Desulfovibrio desulfuricans ATCC 27774, grown with nitrate as the terminal electron acceptor, which is the first example of a bacterioferritin from a strict anaerobic organism. This new bacterioferritin was isolated mainly as a 24-mer of 20 kDa identical subunits, containing 0.5 noncovalently bound heme and 2 iron atoms per monomer. Although its N-terminal sequence is significantly homologous with ferritins from other microorganisms and the ligands to the di-iron ferroxidase center are conserved, it is one of the most divergent bacterioferritins so far characterized. Also, in contrast to all other known bacterioferritins, its heme is not of the B type; its chromatographic behavior is identical to that of iron uroporphyrin. Thus, D. desulfuricans bacterioferritin appears to be the second example of a protein unexpectedly containing this heme cofactor, or a closely related porphyrin, after its finding in Desulfovibrio gigas rubredoxin:oxygen oxidoreductase ?Timkovich, R., Burkhalter, R. S., Xavier, A. V., Chen, L., and Le Gall, J. (1994) Bioorg. Chem. 22, 284-293. The oxidized form of the protein has a visible spectrum characteristic of low-spin ferric hemes, exhibiting a weak absorption band at 715 nm, indicative of bis-methionine heme axial coordination; upon reduction, the alpha-band appears at 550 nm and a splitting of the Soret band occurs, with two maxima at 410 and 425 nm. The heme center has a reduction potential of 140 +/- 10 mV (pH 7.6), a value unusually high compared to that of other bacterioferritins (ca. -200 mV). PMID- 10841765 TI - Localized control of proton transfer through the D-pathway in cytochrome c oxidase: application of the proton-inventory technique. AB - In the reaction cycle of cytochrome c oxidase from Rhodobacter sphaeroides, one of the steps that are coupled to proton pumping, the oxo-ferryl-to-oxidized transition (F --> O), displays a large kinetic deuterium isotope effect of about 7. In this study we have investigated in detail the dependence of the kinetics of this reaction step ?k(FO)(chi) on the fraction (chi) D(2)O in the enzyme solution (proton-inventory technique). According to a simplified version of the Gross Butler equation, from the shape of the graph describing k(FO)(chi)/k(FO)(0), conclusions can be drawn concerning the number of protonatable sites involved in the rate-limiting proton-transfer reaction step. Even though the proton-transfer reaction during the F --> O transition takes place over a distance of at least 30 A and involves a large number of protonatable sites, the proton-inventory analysis displayed a linear dependence, which indicates that the entire deuterium isotope effect of 7 is associated with a single protonatable site. On the basis of experiments with site-directed mutants of cytochrome c oxidase, this localized proton-transfer rate control is proposed to be associated with glutamate (I-286) in the D-pathway. Consequently, the results indicate that proton transfer from the glutamate controls the rate of all events during the F --> O reaction step. The proton-inventory analysis of the overall enzyme turnover reveals a nonlinear plot characteristic of at least two protonatable sites involved in the rate limiting step in the transition state, which indicates that this step does not involve proton transfer through the same pathway (or through the same mechanism) as during the F --> O transition. PMID- 10841766 TI - Stoichiometry of complex formation between Copper(I) and the N-terminal domain of the Menkes protein. AB - The inherent cellular toxicity of copper ions demands that their concentration be carefully controlled. The cellular location of the Menkes ATPase, a key element in the control of intracellular copper, is regulated by the intracellular copper concentration through the N-terminus of the enzyme, comprising 6 homologous subdomains or modules, each approximately 70 residues in length and containing a Cys-X-X-Cys- motif. Based on the proposal that binding of copper to these modules regulates the Menkes ATPase cellular location by promoting changes in the tertiary structure of the enzyme, we have expressed the entire N-terminal domain (MNKr) and the second metal-binding module (MNKr2) of the Menkes protein in E. coli and purified them to homogeneity. Ultraviolet-visible, luminescence, and X ray absorption spectroscopy show that copper and silver bind to the single module, MNKr2, with a stoichiometry of one metal ion per module. However, the array of six modules, MNKr, binds Cu(I) to produce a homogeneous conformer with 4 mol equiv of metal ion. The metal ions are bound in an environment that is shielded from solvent molecules. We suggest a model of the Menkes protein in which the Cu(I) binding induces tertiary changes in the organization of the six metal-binding domains. PMID- 10841767 TI - Ca(2+)- and H(+)-dependent conformational changes of calbindin D(28k). AB - Calbindin D(28k) is a member of a large family of intracellular Ca(2+) binding proteins characterized by EF-hand structural motifs. Some of these proteins are classified as Ca(2+)-sensor proteins, since they are involved in transducing intracellular Ca(2+) signals by exposing a hydrophobic patch on the protein surface in response to Ca(2+) binding. The hydrophobic patch serves as an interaction site for target enzymes. Other members of this group are classified as Ca(2+)-buffering proteins, because they remain closed after Ca(2+) binding and participate in Ca(2+) buffering and transport functions. ANS (8 anilinonaphthalene-1-sulfonic acid) binding and affinity chromatography on a hydrophobic column suggested that both the Ca(2+)-free and Ca(2+)-loaded form of calbindin D(28k) have exposed hydrophobic surfaces. Since exposure of hydrophobic surface is unfavorable in the aqueous intracellular milieu, calbindin D(28k) most likely interacts with other cellular components in vivo. A Ca(2+)-induced conformational change was readily detected by several optical spectroscopic methods. Thus, calbindin D(28k) shares some of the properties of Ca(2+)-sensor proteins. However, the Ca(2+)-induced change in exposed hydrophobic surface was considerably less pronounced than that in calmodulin. The data also shows that calbindin D(28k) undergoes a rapid and reversible conformational change in response to a H(+) concentration increase within the physiological pH range. The pH-dependent conformational change was shown to reside mainly in EF-hands 1-3. Urea-induced unfolding of the protein at pH 6, 7, and 8 showed that the stability of calbindin D(28k) was increased in response to H(+) in the range examined. The results suggest that calbindin D(28k) may interact with targets in a Ca(2+)- and H(+)-dependent manner. PMID- 10841768 TI - Lipid binding of the exchangeable apolipoprotein apolipophorin III induces major changes in fluorescence properties of tryptophans 115 and 130. AB - The effect of lipid association on the local environment of the two tryptophan residues of Locusta migratoria apolipophorin III (apoLp-III) has been studied. In the lipid-free state, Trp115 in helix 4 is buried in the hydrophobic interior of the helix bundle, while Trp130 is located in a loop connecting helices 4 and 5. Fluorescence spectroscopy of single Trp mutants revealed an emission maximum (lambda(max)) of 321 nm for apoLp-III-W@115 (excitation 280 nm) which red-shifted to 327 nm upon binding to dimyristoylphosphatidylcholine (DMPC). ApoLp-III-W@130 displayed a lambda(max) of 338 nm while interaction with DMPC resulted in a blue shift to 331 nm. Quenching studies with KI and acrylamide revealed decreased accessibility to Trp115 compared to Trp130, while lipid binding induced a decrease in quenching of Trp130. Aromatic circular dichroism (CD) spectra showed that Trp vibronic transitions at 278, 286, and 294 nm for lipid-free apoLp-III were caused by Trp115. Upon lipid association, aromatic extrema are reversed in sign, becoming entirely negative with both Trp residues contributing to the vibronic transitions, implying restriction in side-chain mobility of these residues. Thus, lambda(max), quencher accessibility, and aromatic CD analysis indicate that Trp115 is much less solvent-exposed than Trp130. Differences in fluorescence properties of these residues are minimized in the lipid-bound state, a result of relocation of Trp115 and Trp130 into the lipid milieu. Thus, in addition to the hydrophobic faces of apoLp-III amphipathic alpha-helices, the loop region containing Trp130 comes in close contact with DMPC. PMID- 10841769 TI - Paramecium calmodulin mutants defective in ion channel regulation can bind calcium and undergo calcium-induced conformational switching. AB - Calmodulin (CaM) is an essential eukaryotic protein that binds calcium ions cooperatively at four EF-hand binding sites to regulate signal transduction pathways. Interactions between the apo domains of vertebrate CaM reduce the calcium affinities of sites I and II below their intrinsic values, allowing sequential opening of the two hydrophobic clefts in CaM. Viable domain-specific mutants of Parameciumcalmodulin (PCaM) differentially affect ion channels and provide a unique opportunity to dissect the roles of the two highly homologous half-molecule domains. Calcium binding induced an increase in the level of ordered secondary structure and a decrease in Stokes radius in these mutants; such changes were identical in direction to those of wild type CaM, but the magnitude depended on the mutation. Calcium titrations monitored by changes in the intrinsic fluorescence of Y138 in site IV showed that the affinities of sites III and IV of wild type PCaM were (i) higher than those of the same sites in rat CaM, (ii) equivalent to those of the same sites in PCaM mutants altered between sites I and II, and (iii) higher than those of PCaM mutants modified in sites III and IV. Thus, calcium saturation drove all mutants to undergo conformational switching in the same direction but not to the same extent as wild type PCaM. The disruption of the allosteric mechanism that is manifest as faulty channel regulation may be explained by altered properties of switching among the 14 possible partially saturated species of PCaM rather than by an inability to adopt two end-state conformations or target interactions similar to those of the wild type protein. PMID- 10841770 TI - Independent functions for the N- and C-termini in the overlap region of tropomyosin. AB - Tropomyosin (TM) is a coiled-coil that binds head-to-tail along the helical actin filament. The ends of 284-residue tropomyosins are believed to overlap by about nine amino acids. The present study investigates the function of the N- and C terminal overlap regions. Recombinant tropomyosins were produced in Escherichia coli in which nine amino acids were truncated from the N-terminal, C-terminal, or both ends of striated muscle alpha-tropomyosin (TM9a) and TM2 (TM9d), a nonmuscle alpha-tropomyosin expressed in many cells. The two isoforms are identical except for the C-terminal 27 amino acids encoded by exon 9a (striated) or exon 9d (TM2). Removal of either end greatly reduces the actin affinity of both tropomyosins in all conditions and the cooperativity with which myosin promotes tropomyosin binding to actin in the open state. N-Terminal truncations generally are more deleterious than C-terminal truncations. With TM9d, truncation of the N-terminus is as deleterious as both for myosin S1-induced binding. None of the TM9d variants binds well to actin with troponin (+/-Ca(2+)). TM9a with the truncated N terminus binds more weakly to actin with troponin (-Ca(2+)) than when the C terminus is removed but more strongly than when both ends are removed; the actin binding of all three forms is cooperative. The results show that the ends of TM9a, though important, are not required for cooperative function and suggest they have independent functions beyond formation of an overlap complex. The nonadditivity of the TM9d truncations suggests that the ends may primarily function as a complex in this isoform. A surprising result is that all variants bound with the same affinity, and noncooperatively, to actin saturated with myosin S1. Evidently, end-to-end interactions are not required for high-affinity binding to acto-myosin S1. PMID- 10841771 TI - A limited spectrum of mutations causes constitutive activation of the yeast alpha factor receptor. AB - Activation of G protein coupled receptors (GPCRs) by binding of ligand is the initial event in diverse cellular signaling pathways. To examine the frequency and diversity of mutations that cause constitutive activation of one particular GPCR, the yeast alpha-factor receptor, we screened libraries of random mutations for constitutive alleles. In initial screens for mutant receptor alleles that exhibit signaling in the absence of added ligand, 14 different point mutations were isolated. All of these 14 mutants could be further activated by alpha factor. Ten of the mutants also acquired the ability to signal in response to binding of desTrp(1)?Ala(3)alpha-factor, a peptide that acts as an antagonist toward normal alpha-factor receptors. Of these 10 mutants, at least eight alleles residing in the third, fifth, sixth, and seventh transmembrane segments exhibit bona fide constitutive signaling. The remaining alleles are hypersensitive to alpha-factor rather than constitutive. They can be activated by low concentrations of endogenous alpha-factor present in MATa cells. The strongest constitutively active receptor alleles were recovered multiple times from the mutational libraries, and extensive mutagenesis of certain regions of the alpha factor receptor did not lead to recovery of any additional constitutive alleles. Thus, only a limited number of mutations is capable of causing constitutive activation of this receptor. Constitutive and hypersensitive signaling by the mutant receptors is partially suppressed by coexpression of normal receptors, consistent with preferential association of the G protein with unactivated receptors. PMID- 10841772 TI - Prenyl-flavonoids as potent inhibitors of the Pdr5p multidrug ABC transporter from Saccharomyces cerevisiae. AB - The Pdr5p multidrug ABC ("ATP-binding cassette) transporter was highly overexpressed in plasma membranes from a yeast strain exhibiting both pdr1-3 gain of-function mutation in the transcription factor-encoding gene PDR1 and disruption of genes encoding other plasma membrane ABC transporters. Solubilized and purified Pdr5p displayed a tryptophan-characteristic intrinsic fluorescence, whose quenching was used to monitor interactions with substrates and effectors. The transporter exhibited a magnesium-dependent binding affinity for ATP and its fluorescent analogue 2'(3')-N-methylanthraniloyl-ATP, producing a marked fluorescence resonance-energy transfer. It also bound a series of known drug substrates and modulators. Interestingly, yeast Pdr5p interacted with flavonoids recently found to bind to cancer cell P-glycoprotein and to the protozoan parasite multidrug transporter. The extent of high-affinity binding of prenyl flavonoids to purified Pdr5p was correlated to their efficiency to inhibit energy dependent quenching of rhodamine 6G fluorescence catalyzed by Pdr5p-enriched plasma membranes. The hydrophobic flavonoid derivative 6-(3, 3 dimethylallyl)galangin was the most efficient, with a K(i) of 0.18 microM for competitive inhibition of the MgATP-dependent quenching of rhodamine 6G fluorescence. In contrast, inhibition of either ATP or UTP hydrolysis occurred at much higher concentrations and appeared to be noncompetitive. Prenyl-flavonoids therefore behave as potent inhibitors of drug binding to the yeast Pdr5p ABC transporter. PMID- 10841773 TI - Nitric oxide inhibition of free radical-mediated cholesterol peroxidation in liposomal membranes. AB - The ability of nitric oxide ((*)NO) to inhibit propagative lipid peroxidation was investigated using unilamellar liposomes (LUVs) constituted with egg phosphatidylcholine (PC) or 1-palmitoyl-2-oleoylphosphatidylcholine (POPC), [(14)C]cholesterol (Ch), and a nonregenerable singlet oxygen-derived primer, 5alpha-hydroperoxycholesterol (5alpha-OOH). Exposing LUVs to ascorbate and a lipophilic iron chelate at 37 degrees C resulted in an exponential decay of 5alpha-OOH and accumulation of free radical-derived 7alpha- and 7beta hydroperoxycholesterol (7alphabeta-OOH), as detected by high-performance liquid chromatography with electrochemical detection. Thiobarbituric acid-reactive species (TBARS) were generated concurrently in egg PC-containing LUVs. Including the (*)NO donor spermine NONOate (SPNO, 5-50 microM) or S-nitroso-N-acetyl-D,L penicillamine (SNAP, 50-100 microM) in the reaction mixture had no effect on 5alpha-OOH decay (suggesting that iron was not redox-inhibited) but slowed TBARS and 7alphabeta-OOH accumulation in a strongly dose-dependent fashion. Decomposed SPNO or SNAP had no such effects, implying that (*)NO was the responsible agent. Accumulation of several [(14)C]Ch oxidation products, detected by high performance thin-layer chromatography with phosphorimaging, was similarly diminished by active SPNO or SNAP. Concomitantly, a new band referred to as RCh.4 appeared, the radioactivity of which increased as a function of incubation time and (*)NO donor concentration. RCh.4 material was also generated via direct iron/ascorbate reduction of 7alpha-OOH in the presence of (*)NO, consistent with 7alpha-nitrite (7alpha-ONO) identity. However, various other lines of evidence suggest that RCh.4 is not 7alpha-ONO, but rather 5alpha-hydroxycholesterol (5alpha-OH) generated by reduction of 5alpha-ONO arising from 7alpha-ONO rearrangement. 5alpha-OH was only detected when (*)NO was present in the reaction system, thus providing indirect evidence for the existence of nitrosated Ch intermediates arising from (*)NO chain-breaking activity. PMID- 10841774 TI - Oxidative stress and vanadate induce tyrosine phosphorylation of phosphoinositide dependent kinase 1 (PDK1). AB - Phosphoinositide-dependent kinase (PDK1) regulates a number of pathways involved in responses to stress and in growth factor signaling; however, little is known concerning the mechanisms governing the activity of PDK1. In this report, we find that oxidative stress (H(2)O(2)) and vanadate induce tyrosine phosphorylation of PDK1. These effects of H(2)O(2) and vanadate were found in 293T cells and CH310T1/2 cells expressing exogenous PDK1 and in A20 lymphoma cells expressing endogenous PDK1. Exogenously expressed PDK1 was also tyrosine-phosphorylated in response to NGF treatment of 293T expressing TrkA. H(2)O(2) induced a more rapid tyrosine phosphorylation of PDK1 relative to vanadate, and only vanadate-induced tyrosine phosphorylation of PDK1 was sensitive to pretreatment of cells with wortmannin. In vitro, PDK1 could be tyrosine-phosphorylated by both the c-Src and Abl tyrosine kinases. Both H(2)O(2) and vanadate treatments increased the activity of PDK1 when the serum/glucocorticoid regulated kinase (SGK) was used as substrate. Vanadate treatment appeared to bypass the requirement for phosphatidylinositol 3,4,5-trisphosphate when Akt was used as substrate for PDK1. Tyrosine phosphorylation of PDK1 by the Abl tyrosine kinase also increased the activity of PDK1 toward SGK and Akt. These data suggest a novel mechanism through which PDK1 activity may be regulated. PMID- 10841775 TI - Site-directed mutagenesis of human endothelial cell ecto-ADPase/soluble CD39: requirement of glutamate 174 and serine 218 for enzyme activity and inhibition of platelet recruitment. AB - Endothelial cell CD39/ecto-ADPase plays a major role in vascular homeostasis. It rapidly metabolizes ADP released from stimulated platelets, thereby preventing further platelet activation and recruitment. We recently developed a recombinant, soluble form of human CD39, solCD39, with enzymatic and biological properties identical to CD39. To identify amino acids essential for enzymatic/biological activity, we performed site-directed mutagenesis within the four highly conserved apyrase regions of solCD39. Mutation of glutamate 174 to alanine (E174A) and serine 218 to alanine (S218A) resulted in complete and approximately 90% loss of solCD39 enzymatic activity, respectively. Furthermore, compared to wild-type, S57A exhibited a 2-fold increase in ADPase activity without change in ATPase activity, while the tyrosine 127 to alanine (Y127A) mutant lost 50-60% of both ADPase and ATPase activity. The ADPase activity of wild-type solCD39 and each mutant, except for R135A, was greater with calcium as the required divalent cation than with magnesium, but for ATPase activity generally no such preference was observed. Y127A demonstrated the highest calcium/magnesium ADPase activity ratio, 2.8-fold higher than that of wild-type, even though its enzyme activity was greatly reduced. SolCD39 mutants were further characterized by correlating enzymatic with biological activity in an in vitro platelet aggregation system. Each solCD39 mutant was similar to wild-type in reversing platelet aggregation, except for E174A and S218A. E174A, completely devoid of enzymatic activity, failed to inhibit platelet responsiveness, as anticipated. S218A, with 91% loss of ADPase activity, could still reverse platelet aggregation, albeit much less effectively than wild-type solCD39. Thus, glutamate 174 and serine 218 are essential for both the enzymatic and biological activity of solCD39. PMID- 10841776 TI - Interaction of human thiol-specific antioxidant protein 1 with erythrocyte plasma membrane. AB - During the purification from human erythrocytes, human thiol-specific antioxidant protein 1 (hTSA1), one human member of the TSA/alkyl hydroperoxide reductase subunit C (AhpC) family, was fragmented to a molecular mass of 20 323.9300. The fragmented form, in contrast to the intact form, did not bind to the C-terminal peptide (Gln-185-Gln-197) antibody. On the basis of the molecular mass of the fragmented form, the cleavage site was calculated to be between Val-186 and Asp 187. The C-terminal region of hTSA1 appeared to be unnecessary for the antioxidant reaction. In addition to hTSA1, two isoenzymes (hORF06 and hTSA2) were detected in the soluble fraction, whereas only hTSA1 was detected in the membrane fraction. A membrane binding study shows that the intact form binds to erythrocyte plasma membrane but the fragment does not, which suggests that the deleted C-terminal legion (Asp-187-Gln-197) is required for the membrane binding. A model membrane study using phospholipid vesicle showed a strong association of hTSA1 with the phospholipid. Human TSA1 exhibited high catalytic activity for the reduction of the fatty acid hydroperoxide as indicated by K(m) and V(max) (89.9 microM for linoleic acid hydroperoxide, 28.64 micromol(-1) min(-1) mg(-1), respectively). In this paper, we are making the first report of the involvement of the C-terminal region of hTSA1 in membrane binding as evidence supporting the existence of the membrane-associated forms in the erythrocyte. On the basis of our observations, we suggest that hTSA1 can act as a very effective antioxidant to remove oxidative stresses not only in matrix as a free form but also in the membrane surface of red blood cells (RBC) as a membrane-associated form. PMID- 10841777 TI - Oxidative stress induces intracellular accumulation of amyloid beta-protein (Abeta) in human neuroblastoma cells. AB - Several lines of evidence suggest that enhanced oxidative stress is involved in the pathogenesis and/or progression of Alzheimer's disease (AD). Amyloid beta protein (Abeta) that composes senile plaques, a major neuropathological hallmark of AD, is considered to have a causal role in AD. Thus, we have studied the effect of oxidative stress on Abeta metabolism within the cell. Here, we report that oxidative stress induced by H(2)O(2) (100-250 microM) caused an increase in the levels of intracellular Abeta in human neuroblastoma SH-SY5Y cells. Treatment with 200 microM H(2)O(2) caused significant decreases in the protein levels of full-length beta-amyloid precursor protein (APP) and its COOH-terminal fragment that is generated by beta-cleavage, while the gene expression of APP was not altered under these conditions. A pulse-chase experiment further showed a decrease in the half-life of this amyloidogenic COOH-terminal fragment but not in that of nonamyloidogenic counterpart in the H(2)O(2)-treated cells. These results suggest that oxidative stress promotes intracellular accumulation of Abeta through enhancing the amyloidogenic pathway. PMID- 10841778 TI - Denaturation of phosphofructokinase-1 from Saccharomyces cerevisiae by guanidinium chloride and reconstitution of the unfolded subunits to their catalytically active form. AB - Unfolding and refolding of heterooctameric phosphofructokinase-1 from Saccharomyces cerevisiae were investigated by application of kinetic, hydrodynamic, and spectroscopic methods and by use of guanidinium chloride (GdmCl) as denaturant. Inactivation of the enzyme starts at about 0.3 M GdmCl and undergoes a sharp unfolding transition in a narrow range of the denaturant concentration. The inactivation is accompanied by a dissociation of the enzyme into dimers (at 0.6 M GdmCl), which could be detected by changes of the circular dichroism and intrinsic fluorescence. Protein aggregates were observed from 0.7 to 1.5 M GdmCl that unfold at higher denaturant concentrations. Refolding of chemically denatured phosphofructokinase proceeds as a stepwise process via the generation of elements of secondary structure, the formation of assembly competent monomers that associate to heterodimers and the assembly of dimers to heterotetramers and heterooctamers. The assembly reactions seem to be rate limiting. Recovery of the enzyme activity (maximum 65%) competes with an nonproductive aggregation of the subunits. alpha-Cyclodextrin functions as an artificial chaperone by preventing aggregation of the subunits, whereas ATP is suggested to support the generation of heterodimers that are competent to a further assembly. PMID- 10841779 TI - Catalytic cysteine of thymidylate synthase is activated upon substrate binding. AB - The role of Ser 167 of Escherichia coli thymidylate synthase (TS) in catalysis has been characterized by kinetic and crystallographic studies. Position 167 variants including S167A, S167N, S167D, S167C, S167G, S167L, S167T, and S167V were generated by site-directed mutagenesis. Only S167A, S167G, S167T, and S167C complemented the growth of thymidine auxotrophs of E. coli in medium lacking thymidine. Steady-state kinetic analysis revealed that mutant enzymes exhibited k(cat) values 1.1-95-fold lower than that of the wild-type enzyme. Relative to wild-type TS, K(m) values of the mutant enzymes for 2'-deoxyuridylate (dUMP) were 5-90 times higher, while K(m) values for 5,10-methylenetetrahydrofolate (CH(2)H(4)folate) were 1.5-16-fold higher. The rate of dehalogenation of 5-bromo 2'-deoxyuridine 5'-monophosphate (BrdUMP), a reaction catalyzed by TS that does not require CH(2)H(4)folate as cosubstrate, by mutant TSs was analyzed and showed that only S167A and S167G catalyzed the dehalogenation reaction and values of k(cat)/K(m) for the mutant enzymes were decreased by 10- and 3000-fold, respectively. Analysis of pre-steady-state kinetics of ternary complex formation revealed that the productive binding of CH(2)H(4)folate is weaker to mutant TSs than to the wild-type enzyme. Chemical transformation constants (k(chem)) for the mutant enzymes were lower by 1.1-6.0-fold relative to the wild-type enzyme. S167A, S167T, and S167C crystallized in the I2(1)3 space group and scattered X rays to either 1.7 A (S167A and S167T) or 2.6 A (S167C). The high-resolution data sets were refined to a R(crys) of 19.9%. In the crystals some cysteine residues were derivatized with 2-mercaptoethanol to form S,S-(2-hydroxyethyl)thiocysteine. The pattern of derivatization indicates that in the absence of bound substrate the catalytic cysteine is not more reactive than other cysteines. It is proposed that the catalytic cysteine is activated by substrate binding by a proton transfer mechanism in which the phosphate group of the nucleotide neutralizes the charge of Arg 126', facilitating the transfer of a proton from the catalytic cysteine to a His 207-Asp 205 diad via a system of ordered water molecules. PMID- 10841780 TI - Structural differences in the two agonist binding sites of the Torpedo nicotinic acetylcholine receptor revealed by time-resolved fluorescence spectroscopy. AB - The nicotinic acetylcholine receptor (nAChR) from Torpedo marmorata carries two nonequivalent agonist binding sites at the alphadelta and alphagamma subunit interfaces. These sites have been characterized by time-resolved fluorescence with the partial nicotinic agonist dansyl-C(6)-choline (Dnscho). When bound to the detergent-solubilized receptor, the fluorescence lifetime distribution of Dnscho displays a characteristic signature, with four separable components at 0.2, 1.8, 7.2, and 18.3 ns, respectively. Competition experiments with the antagonist d-tubocurarine (dTC), known to bind preferentially to the alphagamma site, result in substantial changes of this signature, associated with a strong decrease in average fluorescence lifetime. Comparisons with two other competitive antagonists, alpha-conotoxin M1 and alpha-bungarotoxin, demonstrate that Dnscho binds with a similar affinity to the two sites but that the microenvironment of the probe is different for each site. Using a two-site binding model together with published equilibrium constants to describe the competitive binding of dTC and Dnscho, we reach a satisfactory description of the changes in fluorescence lifetimes and propose characteristic fluorescence parameters of the probe bound to each type of site. This analysis indicates that Dnscho at the alphadelta site is principally associated with a 8.7 ns lifetime, while it has a 20.2 ns major lifetime at the alphagamma site. Therefore, the observed fluorescence heterogeneity arises in large part from the structural differences of the two binding sites. As a result, this signal can be used to identify the binding preferences of competitive ligands of unknown pharmacology. PMID- 10841781 TI - Targeting a Pneumocystis carinii group I intron with methylphosphonate oligonucleotides: backbone charge is not required for binding or reactivity. AB - Pneumocystis carinii is a mammalian pathogen that contains a self-splicing group I intron in its large subunit rRNA precursor. We report the binding of methylphosphonate/DNA chimeras and neutral methylphosphonate oligonucleotides to a ribozyme that is a truncated form of the intron. At 15 mM Mg(2+), the nuclease resistant all-methylphosphonate hexamer, d(AmTmGmAmCm)rU, with a sequence that mimics the 3' end of the precursor's 5' exon, binds with a dissociation constant of 272 nM. The hexamer's dissociation constant for binding by base-pairing alone to the ribozyme's binding site sequence is 8.3 mM. Thus there is a 30 000-fold binding enhancement by tertiary interactions (BETI), which is close to the 60 000 fold enhancement previously observed with the all-ribo hexamer, r(AUGACU). Evidently, backbone charge and 2' hydroxyl groups are not required for BETI. At 3 15 mM Mg(2+), the all-methylphosphonate and DNA oligonucleotides trans-splice to a truncated form of the rRNA precursor, but do not compete with cis-splicing when pG is present. These results suggest that uncharged or partially charged backbones may be used to design therapeutics to target RNAs through binding enhancement by tertiary interactions and suicide inhibition strategies. PMID- 10841782 TI - Orientation of coenzyme A substrates, nicotinamide and active site functional groups in (Di)enoyl-coenzyme A reductases. AB - The stereochemical course of reduction of dienoyl-coenzyme A (CoA) thiolesters catalyzed by the 2,4-dienoyl-CoA reductase from rat liver mitochondria was investigated. The configuration of the double bond in the 3-enoyl-CoA products was determined by (1)H NMR, and experiments to determine the stereochemical course of reduction at Calpha and Cdelta by use of 4-(2)H-labeled beta nicotinamide adenine dinucleotide phosphate, reduced form (NADPH), were conducted in H(2)O and D(2)O. Defining the diastereoselectivity of the reaction, catalyzed by the Delta(3),Delta(2)-enoyl-CoA isomerase, facilitated the determination of the stereochemical course of reduction by 2, 4-dienoyl-CoA reductase. The absence of solvent exchange of the proton transferred during the Delta(3),Delta(2)-enoyl CoA isomerase catalyzed equilibration of trans-2- and trans-3-enoyl-CoAs, coupled with the strong sequence homology to enoyl-CoA hydratase support the intramolecular suprafacial transfer of the pro-2R proton of trans-3-enoyl-CoA to the pro-4R position of trans-2-enoyl-CoA. The results indicate that the configuration of the double bond of the 3-enoyl-CoA product is trans and that a general acid-catalyzed addition of a solvent derived proton/deuteron occurs on the si face at Calpha of the dienoyl-CoA. The addition of the pro-4S hydrogen from NADPH occurs on the si face at Cdelta of trans-2, cis-4-dienoyl-CoA and on the re face at Cdelta of trans-2, trans-4-dienoyl-CoA. The stereochemical course of reduction of InhA, an enoyl-thiolester reductase from Mycobacterium tuberculosis, was also determined by use of ?4-(2)HNADH in D(2)O. The reduction of trans-2-octenoyl-CoA catalyzed by InhA resulted in the syn addition of (2)H(2) across the double bond yielding (2R,3S)-?2, 3-(2)H(2)octanoyl-CoA. In the crystal structure of the InhA ternary complex, the residue donating the proton to Calpha could not be identified ?Rozwarski, D. A., Vilcheze, C., Sugantino, M., Bittman, R., and Sacchettini, J. C. (1999) J. Biol. Chem. 274, 15582-15589. The current results place further restrictions on the source of the proton and suggest the reduction is stepwise. PMID- 10841783 TI - The first structure of UDP-glucose dehydrogenase reveals the catalytic residues necessary for the two-fold oxidation. AB - Bacterial UDP-glucose dehydrogenase (UDPGlcDH) is essential for formation of the antiphagocytic capsule that protects many virulent bacteria such as Streptococcus pyogenes andStreptococcus pneumoniae type 3 from the host's immune system. We have determined the X-ray structures of both native and Cys260Ser UDPGlcDH from S. pyogenes (74% similarity to S. pneumoniae) in ternary complexes with UDP xylose/NAD(+) and UDP-glucuronic acid/NAD(H), respectively. The 402 residue homodimeric UDPGlcDH is composed of an N-terminal NAD(+) dinucleotide binding domain and a C-terminal UDP-sugar binding domain connected by a long (48 A) central alpha-helix. The first 290 residues of UDPGlcDH share structural homology with 6-phosphogluconate dehydrogenase, including conservation of an active site lysine and asparagine that are implicated in the enzyme mechanism. Also proposed to participate in the catalytic mechanism are a threonine and a glutamate that hydrogen bond to a conserved active site water molecule suitably positioned for general acid/base catalysis. PMID- 10841784 TI - Metal binding modes of Alzheimer's amyloid beta-peptide in insoluble aggregates and soluble complexes. AB - Aggregation of the amyloid beta-peptide (Abeta) into insoluble fibrils is a key pathological event in Alzheimer's disease. Zn(II) induces the Abeta aggregation at acidic-to-neutral pH, while Cu(II) is an effective inducer only at mildly acidic pH. We have examined Zn(II) and Cu(II) binding modes of Abeta and their pH dependence by Raman spectroscopy. The Raman spectra clearly demonstrate that three histidine residues in the N-terminal hydrophilic region provide primary metal binding sites and the solubility of the metal-Abeta complex is correlated with the metal binding mode. Zn(II) binds to the N(tau) atom of the histidine imidazole ring and the peptide aggregates through intermolecular His(N(tau)) Zn(II)-His(N(tau)) bridges. The N(tau)-metal ligation also occurs in Cu(II) induced Abeta aggregation at mildly acidic pH. At neutral pH, however, Cu(II) binds to N(pi), the other nitrogen of the histidine imidazole ring, and to deprotonated amide nitrogens of the peptide main chain. The chelation of Cu(II) by histidine and main-chain amide groups results in soluble Cu(II)-Abeta complexes. Under normal physiological conditions, Cu(II) is expected to protect Abeta against Zn(II)-induced aggregation by competing with Zn(II) for histidine residues of Abeta. PMID- 10841785 TI - Disulfide bonds and protein folding PMID- 10841786 TI - SPR biosensor studies of the direct interaction between 27 drugs and a liposome surface: correlation with fraction absorbed in humans. PMID- 10841787 TI - Non-amidine-containing 1,2-dibenzamidobenzene inhibitors of human factor Xa with potent anticoagulant and antithrombotic activity. PMID- 10841788 TI - Mapping the active site of angiotensin-converting enzyme by transferred NOE spectroscopy. AB - The interaction of five furylacryloyl (fa)-amino acid derivatives, fa-Phe, fa-Phe Phe, fa-Gly-Leu-NH(2), fa-Ala-Lys, and fa-Trp, with angiotensin-converting enzyme (ACE), a protein of MW = 130 kDa, was studied by transferred NOESY experiments. Identification of fa derivatives binding to ACE as well as determination of their relative affinities could be accomplished directly from the compound mixtures. Of the five fa derivatives we found that fa-Phe, fa-Trp, and fa-Gly-Leu-NH(2) bind more strongly to ACE than the other two. The dissociation constant of fa-Phe was determined from NMR spectra to 5 x 10(-4) M. A large excess of dipeptides competitively displaced fa-Trp and fa-Phe-Phe from the receptor pocket, allowing the binding site to be mapped. Also, the relative affinities of the fa-Phe, fa Ala-Lys, and fa-Gly-Leu-NH(2) changed after addition of the dipeptides with fa Gly-Leu-NH(2) showing the strongest binding. In addition, the presence of a strong inhibitor of the S1' and S2' sites, namely captopril, resulted in the same transferred NOE intensities of fa-Phe, indicating that it binds solely to the S1 and S2 subsites. A rapid screening of binding specificity from mixtures is possible by using a large excess of ligand(s) in transferred NOE studies, even when relatively small amounts of protein are present. PMID- 10841789 TI - Developing a dynamic pharmacophore model for HIV-1 integrase. AB - We present the first receptor-based pharmacophore model for HIV-1 integrase. The development of "dynamic" pharmacophore models is a new method that accounts for the inherent flexibility of the active site and aims to reduce the entropic penalties associated with binding a ligand. Furthermore, this new drug discovery method overcomes the limitation of an incomplete crystal structure of the target protein. A molecular dynamics (MD) simulation describes the flexibility of the uncomplexed protein. Many conformational models of the protein are saved from the MD simulations and used in a series of multi-unit search for interacting conformers (MUSIC) simulations. MUSIC is a multiple-copy minimization method, available in the BOSS program; it is used to determine binding regions for probe molecules containing functional groups that complement the active site. All protein conformations from the MD are overlaid, and conserved binding regions for the probe molecules are identified. Those conserved binding regions define the dynamic pharmacophore model. Here, the dynamic model is compared to known inhibitors of the integrase as well as a three-point, ligand-based pharmacophore model from the literature. Also, a "static" pharmacophore model was determined in the standard fashion, using a single crystal structure. Inhibitors thought to bind in the active site of HIV-1 integrase fit the dynamic model but not the static model. Finally, we have identified a set of compounds from the Available Chemicals Directory that fit the dynamic pharmacophore model, and experimental testing of the compounds has confirmed several new inhibitors. PMID- 10841790 TI - Synthesis, molecular modeling, and opioid receptor affinity of 9, 10 diazatricyclo[4.2.1.1(2,5)]decanes and 2,7-diazatricyclo[4.4.0. 0(3,8)]decanes structurally related to 3,8-diazabicyclo[3.2. 1]octanes. AB - Various lines of evidence, including molecular modeling studies, imply that the endoethylenic bridge of 3,8-diazabicyclo[3.2. 1]octanes (DBO, 1) plays an essential role in modulating affinity toward mu opioid receptors. This hypothesis, together with the remarkable analgesic properties observed for N(3) propionyl, N(8) arylpropenyl derivatives (2) and of the reverted isomers (3), has prompted us to insert an additional endoethylenic bridge on the piperazine moiety in order to identify derivatives with increased potency toward this receptor class. In the present report, we describe the synthesis of the novel compounds 9,10-diazatricyclo[4.2. 1.1(2,5)]decane (4) and 2,7 diazatricyclo[4.4.0.0(3,8)]decane (5), as well as the representative derivatives functionalized at the two nitrogen atoms by propionyl and arylpropenyl groups (6a e, 7a-d). Opioid receptor binding assays revealed that, among the compounds tested, the N-propionyl-N-cinnamyl derivatives 6a and 7a exhibited the highest mu receptor affinity, and remarkably, compound 7a displayed in vivo (mice) an analgesic potency 6-fold that of morphine. PMID- 10841791 TI - Modeling of kappa-opioid receptor/agonists interactions using pharmacophore-based and docking simulations. AB - The interaction of the kappa-opioid receptor with arylacetamide and benzomorphan derivatives acting as agonists was modeled through pharmacophore-based and docking calculations. Potentially bioactive conformations of representative ligands (U-50,488 and its benzo-fused analogues 4 and 6 for arylacetamides and MPCB for benzomorphans) were identified by systematic conformational analysis and docked into a 3D model of the kappa-receptor. The obtained complexes, refined by energy-minimization and molecular dynamics, were evaluated for their consistency with structure-activity relationships and site-directed mutagenesis data. The following interactions are hypothesized to govern the ligand-receptor recognition process: (i) a salt bridge between the Asp138 carboxylate and the protonated nitrogen of the bound agonist; (ii) a hydrogen bond donated by the Tyr312 hydroxyl to the carbonyl oxygen of arylacetamides and MPCB; (iii) hydrophobic interactions established by the dichlorophenyl moiety of arylacetamides and the pendant phenyl ring of MPCB with the surrounding side chains of Tyr312, Leu224, Leu295, and Ala298; (iv) a pi-stacking contact between the Tyr312 side chain and the phenyl ring of arylacetamides; (v) a hydrogen bond linking the His291 imidazole ring to the phenolic hydroxy group featured by typical benzomorphans and the arylacetamides 4 and 6. PMID- 10841792 TI - Peptide and peptide mimetic inhibitors of antigen presentation by HLA-DR class II MHC molecules. Design, structure-activity relationships, and X-ray crystal structures. AB - Molecular features of ligand binding to MHC class II HLA-DR molecules have been elucidated through a combination of peptide structure-activity studies and structure-based drug design, resulting in analogues with nanomolar affinity in binding assays. Stabilization of lead compounds against cathepsin B cleavage by N methylation of noncritical backbone NH groups or by dipeptide mimetic substitutions has generated analogues that compete effectively against protein antigens in cellular assays, resulting in inhibition of T-cell proliferation. Crystal structures of four ternary complexes of different peptide mimetics with the rheumatoid arthritis-linked MHC DRB10401 and the bacterial superantigen SEB have been obtained. Peptide-sugar hybrids have also been identified using a structure-based design approach in which the sugar residue replaces a dipeptide. These studies illustrate the complementary roles played by phage display library methods, peptide analogue SAR, peptide mimetics substitutions, and structure based drug design in the discovery of inhibitors of antigen presentation by MHC class II HLA-DR molecules. PMID- 10841793 TI - Structure-activity relationships in platelet-activating factor (PAF). 10. From PAF antagonism to inhibition of HIV-1 replication. AB - Excessive levels of PAF and cells of macrophage lineage appear to play an important role in neuronal cell injury, inflammatory syndrome, and HIV replication in CNS resulting in AIDS dementia complex (ADC). The beneficial effects of PAF receptor antagonists are evident and give rise to expected therapeutic strategies for neurotrauma. Piperazine derivatives bearing a "cache oreilles" (ear-muff) electronic distribution are able to inhibit in vitro PAF effects and, thus, could be used in pathologies where this mediator is involved. Therefore, their potential anti-HIV activity was investigated, and we find that (i) these PAF antagonists are effectively active in HIV-infected monocyte-derived macrophages (MDM) but there is no correlation between both anti-HIV and anti-PAF activities; (ii) the presence of a carbamate function (compounds 1a-d) is favorable to the antiviral activity; (iii) the lipophilicity of the substituent on the piperazinic cycle seems to be less important for the anti-PAF activity than for the antiviral one. Our leading compound, PMS 601 (compound 1a), presents a dual activity with IC(50) of 8 and 11 microM for anti-PAF and anti-HIV activity, respectively, without cytotoxic events at 1000 microM in MDM. Although its mode of action is not clearly defined, these data suggest that PMS 601, which displays no effect on acellular reverse transcriptase or protease tests, deserves further investigation in the treatment of HIV-1-associated dementia. PMID- 10841794 TI - Structure-activity relationships in a series of bisquaternary bisphthalimidine derivatives modulating the muscarinic M(2)-receptor allosterically. AB - Hexane-bisammonium-type compounds containing lateral phthalimide moieties are well-established ligands of the common allosteric binding site of muscarinic M(2) receptors. Previous structure-activity relationships (SAR) revealed two positively charged centers and two lateral phthalimide moieties in a defined arrangement to be essential of a high allosteric potency. The purpose of this study was to replace one carbonyl group of the phthalimides with hydrogens, hydroxy, alkoxy, phenyl, benzyl, and benzylidene groups in order to check the influence of these substituents on the allosteric activity in antagonist-linked receptors. The analysis of the quantitative SAR indicated that a high allosteric potency is related to a certain amount of rigidity as well as polarizibility and the ability to form hydrophobic interactions. PMID- 10841795 TI - Synthesis, structure, and neuroprotective properties of novel imidazolyl nitrones. AB - A new series of imidazolyl nitrones spin traps has been synthesized and evaluated pharmacologically. The salient structural feature of these molecules is the presence of an imidazole moiety substituted by aromatic or heteroaromatic cycles. This connectivity imparts to the nitrone superior neuroprotective properties in vivo and in parallel reduced side effects and toxicity. Thus compound 6a (a 2 phenylimidazolyl nitrone) administered intraperitoneally protects (80%) mice from lethality induced by an intracerebroventricular administration of tert-butyl hydroperoxide (t-BHP) an oxidant capable of inducing neurodegenerative processes. Administration of the archetypal nitrone phenyl-tert-butyl nitrone (PBN) at an equimolar dose also affords some protection (60%) in this test. However, this activity is accompanied by hypothermia, whereas no such effect is apparent for 6a. Moreover, previously prepared nonsubstituted or alkyl-substituted imidazolyl nitrones were shown to be extremely toxic to rats in contrast to the compounds prepared in this study. The observed activities in vivo correlate well with the calculated partition coefficients (ClogP) and HOMO energy level. PMID- 10841796 TI - Population, acid-base, and redox properties of N-acetylcysteine conformers. AB - Rotamers of N-acetyl-L-cysteine (NAC, the most popular mucolytic drug) are characterized in terms of populations, site- and conformer-specific acid-base properties, reducing strength, and molecular pharmacology. A new, general relationship between the bulk- and rotamer-specific basicities is introduced. NAC at high pH predominantly exists in a trans thiolate-carboxylate rotameric form, whereas protonation promotes the occurrence of intramolecular hydrogen bond forming isomers. Distribution curves of the rotamers are depicted as a function of pH. Rotamer-dependent thiolate basicities differ by up to 0.5 log k units. Carboxylate basicities show slight conformation-dependence only. The membrane penetrating capabilities from various compartments of the body are assessed on the basis of the pH-dependent charge of the molecule. The thiol-disulfide half cell potential is calculated, using the correlation between the thiolate basicity and oxidizability. The oxidation-reduction properties of NAC are compared to those of other biological thiols in their definite microscopic forms. The pharmacokinetic behavior is interpreted in terms of the physicochemical parameters, providing molecular/submolecular explanation for several therapeutic properties of NAC. PMID- 10841797 TI - Design, synthesis, and structure-activity relationships of phthalimide phenylpiperazines: a novel series of potent and selective alpha(1)(a)-adrenergic receptor antagonists. AB - Beginning from the screening hit and literature alpha(1)-adrenergic compounds, a hybridized basic skeleton A was proposed as the pharmacophore for potent and selective alpha(1a)-AR antagonists. Introduction of a hydroxy group to increase the flexibility afforded B which served as the screening model and resulted in the identification of the second-generation lead 1. Using the Topliss approach, a number of potent and selective alpha(1a)-AR antagonists were discovered. In all cases, binding affinity and selectivity at the alpha(1a)-AR of S-hydroxy enantiomers were higher than those of the R-hydroxy enantiomers. As compared to the des-hydroxy analogues, the S-hydroxy enantiomers had slightly lower binding affinity at alpha(1a)-AR but gained more than 2-fold selectivity for alpha(1a)-AR over alpha(1b)-AR, and 2- to 6-fold selectivity for alpha(1a)-AR over alpha(1d) AR. They also had less cross activities against a panel of 25-35 peripheral and CNS receptors. The S-hydroxy enantiomers 23 and 24 (K(i) = 0.29 nM, 0.33 nM; alpha(1b)/alpha(1a) >5690, >6060; alpha(1d)/alpha(1a) = 186, 158, respectively) were slightly less potent but much more selective at alpha(1a)-AR than tamsulosin (K(i) = 0.13 nM, alpha(1b)/alpha(1a) = 14.8, alpha(1d)/alpha(1a) = 1.4). In the functional assay, the S-hydroxy enantiomers 20, 23, and 24 were less potent than tamsulosin in inhibiting contractions of rat prostate tissue but more selective in the inhibition of tissue contractions of rat prostate versus rat aorta. Compound 24 was selected as the development candidate for the treatment of BPH. PMID- 10841799 TI - Predicting blood-brain barrier permeation from three-dimensional molecular structure. AB - Predicting blood-brain barrier (BBB) permeation remains a challenge in drug design. Since it is impossible to determine experimentally the BBB partitioning of large numbers of preclinical candidates, alternative evaluation methods based on computerized models are desirable. The present study was conducted to demonstrate the value of descriptors derived from 3D molecular fields in estimating the BBB permeation of a large set of compounds and to produce a simple mathematical model suitable for external prediction. The method used (VolSurf) transforms 3D fields into descriptors and correlates them to the experimental permeation by a discriminant partial least squares procedure. The model obtained here correctly predicts more than 90% of the BBB permeation data. By quantifying the favorable and unfavorable contributions of physicochemical and structural properties, it also offers valuable insights for drug design, pharmacological profiling, and screening. The computational procedure is fully automated and quite fast. The method thus appears as a valuable new tool in virtual screening where selection or prioritization of candidates is required from large collections of compounds. PMID- 10841798 TI - Methanocarba analogues of purine nucleosides as potent and selective adenosine receptor agonists. AB - Adenosine receptor agonists have cardioprotective, cerebroprotective, and antiinflammatory properties. We report that a carbocyclic modification of the ribose moiety incorporating ring constraints is a general approach for the design of A(1) and A(3) receptor agonists having favorable pharmacodynamic properties. While simple carbocyclic substitution of adenosine agonists greatly diminishes potency, methanocarba-adenosine analogues have now defined the role of sugar puckering in stabilizing the active adenosine receptor-bound conformation and thereby have allowed identification of a favored isomer. In such analogues a fused cyclopropane moiety constrains the pseudosugar ring of the nucleoside to either a Northern (N) or Southern (S) conformation, as defined in the pseudorotational cycle. In binding assays at A(1), A(2A), and A(3) receptors, (N) methanocarba-adenosine was of higher affinity than the (S)-analogue, particularly at the human A(3) receptor (N/S affinity ratio of 150). (N)-Methanocarba analogues of various N(6)-substituted adenosine derivatives, including cyclopentyl and 3-iodobenzyl, in which the parent compounds are potent agonists at either A(1) or A(3) receptors, respectively, were synthesized. The N(6) cyclopentyl derivatives were A(1) receptor-selective and maintained high efficacy at recombinant human but not rat brain A(1) receptors, as indicated by stimulation of binding of [(35)S]GTP-gamma-S. The (N)-methanocarba-N(6)-(3 iodobenzyl)adenosine and its 2-chloro derivative had K(i) values of 4.1 and 2.2 nM at A(3) receptors, respectively, and were highly selective partial agonists. Partial agonism combined with high functional potency at A(3) receptors (EC(50) < 1 nM) may produce tissue selectivity. In conclusion, as for P2Y(1) receptors, at least three adenosine receptors favor the ribose (N)-conformation. PMID- 10841800 TI - Novel potent ligands for the central nicotinic acetylcholine receptor: synthesis, receptor binding, and 3D-QSAR analysis. AB - In the past few years the focus on central acetylcholine receptors has shifted from compounds with affinity for muscarinic acetylcholine receptors (mAChR) to compounds with affinity for nicotinic acetylcholine receptors (nAChR). The therapeutic potential includes treatment of a variety of diseases, e.g., Alzheimer's disease, Parkinson's disease, and Tourette's syndrome. This work describes the synthesis of six novel series of potent ligands with nanomolar affinity for the alpha4beta2 nAChR subtype. Structure-activity relationship (SAR) was evaluated by the calculation of a 3D-QSAR model. 3D-QSAR analysis of the compounds using the GRID/GOLPE methodology resulted in a model of high quality (R(2) = 0.97, Q(2) = 0.81). The coefficient plots reveal that the steric interactions between the target and our compounds are of major importance for the affinity. Bulky substituents in the 6-position of the pyridine ring will reduce the affinity of the compounds, whereas bulky ring systems including a sp(3) nitrogen will increase the affinity of the compounds. PMID- 10841801 TI - Isoquinoline and quinazoline urea analogues as antagonists for the human adenosine A(3) receptor. AB - Isoquinoline and quinazoline urea derivatives were found to bind to human adenosine A(3) receptors. Series of N-phenyl-N'-quinazolin-4-ylurea derivatives and N-phenyl-N'-isoquinolin-1-ylurea derivatives were synthesized and tested in radioligand binding assays on their adenosine receptor affinities. A structure affinity analysis indicated that on the 2-position of the quinazoline ring or the equivalent 3-position of the isoquinoline ring a phenyl or heteroaryl substituent increased the adenosine A(3) receptor affinity in comparison to unsubstituted or aliphatic derivatives. Furthermore, the structure-affinity relationship of substituted phenylurea analogues was investigated. Substituents such as electron withdrawing or electron-donating groups were introduced at different positions of the benzene ring to probe electronic and positional effects of substitution. Substitution on the 3- or 4-position of the phenyl ring decreased the adenosine A(3) receptor affinity. Substitution at position 2 with an electron-donating substituent, such as methyl or methoxy, increased human adenosine A(3) receptor affinity, whereas substitution on the 2-position with an electron-withdrawing substituent did not influence affinity. Combination of the optimal substituents in the two series had an additive effect, which led to the potent human adenosine A(3) receptor antagonist N-(2-methoxyphenyl)-N'-(2-(3-pyridyl)quinazolin-4 yl)urea (VUF5574, 10a) showing a K(i) value of 4 nM and being at least 2500-fold selective vs A(1) and A(2A) receptors. Compound 10a competitively antagonized the effect of an agonist in a functional A(3) receptor assay, i.e., inhibition of cAMP production in cells expressing the human adenosine A(3) receptor; a pA(2) value of 8.1 was derived from a Schild plot. In conclusion, compound 10a is a potent and selective human adenosine A(3) receptor antagonist and might be a useful tool in further characterization of the human A(3) receptor. PMID- 10841802 TI - Novel inhibitors of nucleoside triphosphate diphosphohydrolases: chemical synthesis and biochemical and pharmacological characterizations. AB - To elucidate the physiological role played by nucleoside triphosphate diphosphohydrolase (NTPDase; EC 3.6.1.5), adenine nucleotide analogues, modified on the purine ring, have been synthesized and tested as potential inhibitors. Resistance of ATP analogues to hydrolysis and their potency as NTPDase inhibitors were evaluated. For this purpose, a particulate fraction isolated from bovine spleen was used as the enzyme source. Among the synthesized analogues, 8 thiobutyladenosine 5'-triphosphate (8-BuS-ATP) was found to be the most effective nonhydrolyzable competitive inhibitor, with an estimated K(i) of 10 microM. This nonhydrolyzable analogue did not exert any P2X-receptor-mediated effect on endothelium-denuded blood vessels, from the guinea pig mesenteric bed. In agreement with this observation, infusion of the analogue did not cause any significant blood pressure variations of the precontracted vessel. Because in previous studies on isolated turkey erythrocytes and rat astrocytes 8-BuS-ATP was not able to trigger any P2Y(1)-receptor-mediated effect, it therefore appears that this NTPDase inhibitor does not interfere with purinergic receptors. PMID- 10841803 TI - Anilides of (R)-trifluoro-2-hydroxy-2-methylpropionic acid as inhibitors of pyruvate dehydrogenase kinase. AB - The optimization of a series of anilide derivatives of (R)-3,3, 3-trifluoro-2 hydroxy-2-methylpropionic acid as inhibitors of pyruvate dehydrogenase kinase (PDHK) is described that started from N-phenyl-3,3,3-trifluoro-2-hydroxy-2 methylpropanamide 1 (IC(50) = 35 +/- 1.4 microM). It was found that small electron-withdrawing groups on the ortho position of the anilide, i.e., chloro, acetyl, or bromo, increased potency 20-40-fold. The oral bioavailability of the compounds in this series is optimal (as measured by AUC) when the anilide is substituted at the 4-position with an electron-withdrawing group (i.e., carboxyl, carboxyamide, and sulfoxyamide). N-(2-Chloro-4-isobutylsulfamoylphenyl)-(R)-3,3, 3-trifluoro-2-hydroxy-2-methylpropionamide (10a) inhibits PDHK in the primary enzymatic assay with an IC(50) of 13 +/- 1.5 nM, enhances the oxidation of [(14)C]lactate into (14)CO(2) in human fibroblasts, lowers blood lactate levels significantly 2.5 and 5 h after oral doses as low as 30 micromol/kg, and increases the ex vivo activity of PDH in muscle, kidney, liver, and heart tissues. However, in contrast to sodium dichloroacetate (DCA), these PDHK inhibitors did not lower blood glucose levels. Nevertheless, they are effective at increasing the utilization and disposal of lactate and could be of utility to ameliorate conditions of inappropriate blood lactate elevation. PMID- 10841804 TI - Synthesis and evaluation of nitroheterocyclic phosphoramidates as hypoxia selective alkylating agents. AB - A series of novel nitroheterocyclic phosphoramidates has been prepared, and the cytotoxicity of these compounds has been evaluated in clonogenic assays against B16, wild-type and cyclophosphamide-resistant MCF-7, and HT-29 cells under aerobic conditions and HT-29 cells under hypoxic conditions. All compounds were comparable in toxicity to wild-type and resistant MCF-7 cells and were also selectively toxic to HT-29 cells under hypoxic conditions (selectivity ratios 1.7 to >20). Analogues lacking the nitro group were not cytotoxic. Electron withdrawing substituents increased cytotoxicity under aerobic conditions and thereby decreased hypoxic selectivity. In contrast, an electron-donating substituent markedly decreased both aerobic and hypoxic cytotoxicity but enhanced hypoxic selectivity. Chemical reduction of the nitro group resulted in rapid expulsion of the cytotoxic phosphoramide mustard. The most potent of these compounds show significant cytotoxicity under both aerobic and hypoxic conditions. PMID- 10841805 TI - Synthesis, in vitro anti-breast cancer activity, and intracellular decomposition of amino acid methyl ester and alkyl amide phosphoramidate monoesters of 3'-azido 3'-deoxythymidine (AZT). AB - We report the synthesis and anticancer activity of a series of AZT phosphoramidate monoesters containing amino acid methyl ester (3a-11a) and N alkyl amide (3b-11b, 9c-9f) moieties. The aromatic amino acid methyl esters were found to be more cytotoxic than the aliphatic analogues toward MCF-7 cells (human pleural effusion breast adenocarcinoma cell line). A marked stereochemical preference for the L-amino acid stereochemistry was also observed in MCF-7 cells. There was no consistent enhancement of cytotoxicity of the methyl amides over the corresponding methyl esters. AZT and the two AZT aromatic amino acid methyl ester phosphoramidates 8a and 9a were found to be more cytotoxic toward MCF-7 cells than to CEM cells (human T-cell lymphoblastic leukemia). The selective cytotoxicity toward MCF-7 cells may be associated with greater intracellular levels of phosphoramidate monoester and/or phosphorylated AZT. PMID- 10841806 TI - Quinolones: novel probes in antifilarial chemotheraphy. AB - Quinolones have been discovered in our laboratory as a new class of antifilarial agents. This has led to the design, synthesis, and antifilarial evaluation of a number of N-substituted quinol-4(1H)-one-3-carboxamide derivatives 4-6. The macrofilaricidal activity of the target compounds was initially evaluated in vivo against Acanthoeilonema viteae by oral administration of 200 mg/kg x 5 days. Among all the synthesized compounds, 13 displayed activity, with the most potent compound (4a) exhibiting 100% macrofilaricidal and 90% microfilaricidal activities. Compound 4e elicited significant macrofilaricidal (80%) response while compound 5c showed 100% sterilization of female worms. Finally, the two most potent macrofilaricidal compounds, namely 4a and 4e, have been screened for their potency against DNA topoisomerase II, and it has been observed that both have the capability to interfere with this enzyme at 10 micromol/mL concentration. The structure-activity relationship (SAR) associated with position 3 and aryl ring substituents is discussed. PMID- 10841807 TI - Modeling cyclooxygenase inhibition. Implication of active site hydration on the selectivity of ketoprofen analogues. AB - Molecular modeling studies performed on the two cyclooxygenase isozymes (COXs) suggest that active site hydration is crucial for understanding inhibitor selectivity. In this work, models have been constructed considering some implicit water molecules, placed in the position suggested by GRID, that participate in the dynamic hydrogen-bonding network at the polar active site entrance together with protein residues 355, 524, 120, and 513. The selectivity observed for ketoprofen (1) and the structural analogues 2 and 3 may be rationalized in terms of such implicit hydration. PMID- 10841808 TI - Topoisomerase I-mediated antiproliferative activity of enantiomerically pure fluorinated homocamptothecins. AB - Homocamptothecin (hCPT) is an E-ring modified camptothecin (CPT) analogue bearing a methylene spacer between the alcohol and carboxyl functions of the CPT lactone. Combining pronounced inhibitory activity of topoisomerase I (Topo I) with enhanced plasma stability, hCPT constitutes an attractive template for the elaboration of new anticancer agents. Fluorinated hCPT analogues, prepared in enantiomerically pure form, were assayed by their stimulation of Topo I-mediated DNA cleavage. Translation into cytotoxicity against tumor cells was evaluated on HT29 human colon adenocarcinoma and on the multidrug resistant lung and bladder tumor cell lines, A549 and T24r. Good correlation is observed between the ability of the drugs to stimulate Topo I-mediated DNA cleavage and the respective 50% inhibitory concentrations (IC(50) values) of the HT29, A549, and T24r cell growth. Fluorine substitution in the A-ring of hCPT was found to have a pronounced influence on biological activity, providing several compounds which are up to 100-fold more potent than CPT in terms of IC(50). Among these, 10,11 difluoro-hCPT has been selected for further development. PMID- 10841809 TI - A new variant of Charcot-Marie-Tooth disease type 2 is probably the result of a mutation in the neurofilament-light gene. AB - Charcot-Marie-Tooth (CMT) disease is the most common inherited motor and sensory neuropathy. The axonal form of the disease is designated as "CMT type 2" (CMT2). Although four loci known to be implicated in autosomal dominant CMT2 have been mapped thus far (on 1p35-p36, 3q13. 1, 3q13-q22, and 7p14), no one causative gene is yet known. A large Russian family with CMT2 was found in the Mordovian Republic (Russia). Affected members had the typical CMT2 phenotype. Additionally, several patients suffered from hyperkeratosis, although the association, if any, between the two disorders is not clear. Linkage with the CMT loci already known (CMT1A, CMT1B, CMT2A, CMT2B, CMT2D, and a number of other CMT-related loci) was excluded. Genomewide screening pinpointed the disease locus in this family to chromosome 8p21, within a 16-cM interval between markers D8S136 and D8S1769. A maximum two-point LOD score of 5.93 was yielded by a microsatellite from the 5' region of the neurofilament-light gene (NF-L). Neurofilament proteins play an important role in axonal structure and are implicated in several neuronal disorders. Screening of affected family members for mutations in the NF-L gene and in the tightly linked neurofilament-medium gene (NF-M) revealed the only DNA alteration linked with the disease: a A998C transversion in the first exon of NF L, which converts a conserved Gln333 amino acid to proline. This alteration was not found in 180 normal chromosomes. Twenty unrelated CMT2 patients, as well as 26 others with an undetermined form of CMT, also were screened for mutations in NF-L, but no additional mutations were found. It is suggested that Gln333Pro represents a rare disease-causing mutation, which results in the CMT2 phenotype. PMID- 10841810 TI - Impaired elastic-fiber assembly by fibroblasts from patients with either Morquio B disease or infantile GM1-gangliosidosis is linked to deficiency in the 67-kD spliced variant of beta-galactosidase. AB - We have previously shown that intracellular trafficking and extracellular assembly of tropoelastin into elastic fibers is facilitated by the 67-kD elastin binding protein identical to an enzymatically inactive, alternatively spliced variant of beta-galactosidase (S-Gal). In the present study, we investigated elastic-fiber assembly in cultures of dermal fibroblasts from patients with either Morquio B disease or GM1-gangliosidosis who bore different mutations of the beta-galactosidase gene. We found that fibroblasts taken from patients with an adult form of GM1-gangliosidosis and from patients with an infantile form, carrying a missense mutations in the beta-galactosidase gene-mutations that caused deficiency in lysosomal beta-galactosidase but not in S-Gal-assembled normal elastic fibers. In contrast, fibroblasts from two cases of infantile GM1 gangliosidosis that bear nonsense mutations of the beta-galactosidase gene, as well as fibroblasts from four patients with Morquio B who had mutations causing deficiency in both forms of beta-galactosidase, did not assemble elastic fibers. We also demonstrated that S-Gal-deficient fibroblasts from patients with either GM1-gangliosidosis or Morquio B can acquire the S-Gal protein, produced by coculturing of Chinese hamster ovary cells permanently transected with S-Gal cDNA, resulting in improved deposition of elastic fibers. The present study provides a novel and natural model validating functional roles of S-Gal in elastogenesis and elucidates an association between impaired elastogenesis and the development of connective-tissue disorders in patients with Morquio B disease and in patients with an infantile form of GM1-gangliosidosis. PMID- 10841811 TI - Segregation analysis of esophageal cancer in a moderately high-incidence area of northern China. AB - In order to explore the mode of inheritance of esophageal cancer in a moderately high-incidence area of northern China, we conducted a pedigree survey on 225 patients affected by esophageal cancer in Yangquan, Shanxi Province. Segregation analysis was performed using the REGTL program of S.A.G.E. The results showed that Mendelian autosomal recessive inheritance of a major gene that influences susceptibility to esophageal cancer provided the best fit to the data. In the best-fitting recessive model, the frequency of the disease allele was.2039. There was a significant sex effect on susceptibility to the disease. The maximum cumulative probability of esophageal cancer among males with the AA genotype was 100%, but, among females, it was 63.5%. The mean age at onset for both men and women was 62 years. The age-dependent penetrances for males with the AA genotype by the ages of 60 and 80 years were 41.6% and 95.2%, respectively, whereas, for females, they were 26.4% and 60.5%, respectively. Incorporating environmental risk factors-such as cigarette smoking, pipe smoking, alcohol drinking, eating hot food, and eating pickled vegetables-into the models did not provide significant improvement of the fit of the models to these data. The results suggest a major locus underlying susceptibility to esophageal cancer with sex specific penetrance. PMID- 10841812 TI - Elastic-fiber pathologies: primary defects in assembly-and secondary disorders in transport and delivery. PMID- 10841813 TI - Comparison of tests for association and linkage in incomplete families. AB - To analyze incomplete families, the following statistical tests can be used: LRAT a simple likelihood-based association test, TRANSMIT, SIBASSOC/STDT, and RCTDT. We compared these four tests, for the diallelic case, on simulated data sets. The comparisons focused on the power to detect linkage and association when different familial structures, resistance to population stratification, resistance to misclassification of the disease status of the healthy sib, and the effect of nonpaternity were considered. The simulations lead to the following conclusions. The type I errors of TRANSMIT, SIBASSOC/STDT, and RCTDT were not affected by population stratification. LRAT showed bias under strong population stratification. High nonpaternity rates can lead to inflated type I errors, highlighting the importance of identification of half sibs. Under different homogeneous models, the power of TRANSMIT was very similar to that of LRAT, and, similarly, no difference in power was observed between SIBASSOC/STDT and RCTDT. Under various recessive and additive models, TRANSMIT was slightly more powerful than SIBASSOC/STDT when monoparental families with one affected and one unaffected sib were analyzed. Under various dominant models, SIBASSOC/STDT was slightly more powerful than TRANSMIT. Misclassification of the disease status of healthy sibs, as well as the discarding of incomplete families, resulted in a consistent loss of power. PMID- 10841814 TI - Combined endocrine therapy for breast cancer--new life for an old idea? PMID- 10841815 TI - Gatekeeper for endometrium: the PTEN tumor suppressor gene. PMID- 10841816 TI - Almost serendipity: alcoholism drug reverses drug resistance in vitro. PMID- 10841817 TI - Plasminogen activator proteins tested as prognostic markers. PMID- 10841818 TI - Stat bite: Incidence of and mortality from cancer of the colon and rectum, 1976 1997. PMID- 10841819 TI - Causes of pain investigated at molecular level. PMID- 10841820 TI - Report to the nation shows cancer mortality, incidence declining. PMID- 10841821 TI - SEER to collect data on underrepresented populations. PMID- 10841822 TI - Alternative remedies for cancer: an update. PMID- 10841823 TI - Markers of DNA repair and susceptibility to cancer in humans: an epidemiologic review. AB - DNA repair is a system of defenses designed to protect the integrity of the genome. Deficiencies in this system likely lead to the development of cancer. The epidemiology of DNA repair capacity and of its effect on cancer susceptibility in humans is, therefore, an important area of investigation. We have summarized all of the published epidemiologic studies on DNA repair in human cancer through 1998 (n = 64) that addressed the association of cancer susceptibility with a putative defect in DNA repair capacity. We have considered study design, subject characteristics, potential biases, confounding variables, and sources of technical variability. Assays of DNA repair capacity used, to date, can be broadly grouped into five categories: 1) tests based on DNA damage induced with chemicals or physical agents, such as the mutagen sensitivity assay, the G(2) radiation assay, induced micronuclei, and the Comet assay; 2) indirect tests of DNA repair, such as unscheduled DNA synthesis; 3) tests based on more direct measures of repair kinetics, such as the host cell reactivation assay; 4) measures of genetic variation associated with DNA repair; and 5) combinations of more than one category of assay. The use of such tests in human populations yielded positive and consistent associations between DNA repair capacity and cancer occurrence (with odds ratios in the range of 1. 4-75.3, with the majority of values between 2 and 10). However, the studies that we have reviewed have limitations, including small sample size, "convenience" controls, the use of cells different from the target organ, and the use of mutagens that do not occur in the natural environment. The evolving ability to study polymorphisms in DNA repair genes may contribute to new understandings about the mechanisms of DNA repair and the way in which DNA repair capacity affects the development of cancer. PMID- 10841824 TI - Blockage of drug resistance in vitro by disulfiram, a drug used to treat alcoholism. AB - BACKGROUND: P-glycoprotein (P-gp) pumps a wide range of cytotoxic drugs out of cells. Inhibiting maturation of P-gp would be a novel method for circumventing P gp-mediated multidrug resistance, which complicates cancer chemotherapy and treatment of patients infected with human immunodeficiency virus. We examined the effect of disulfiram (Antabuse(TM)) on the maturation and activity of P-gp. METHODS: Embryonic kidney cells were transfected with a complementary DNA for the P-pg gene, and the effects of disulfiram on the sensitivity of the transfected cells to cytotoxic agents were determined. Enzyme assays were used to determine the effects of disulfiram on the verapamil-stimulated adenosine triphosphatase (ATPase) activity of P-gp. Disulfiram modifies cysteine residues, and mutant forms of P-gp that lack individual cysteines were used to determine whether particular cysteine residues mediate disulfiram's effects on P-gp activity. Maturation of recombinant P-gp was followed on immunoblots. RESULTS: Disulfiram increased the sensitivity of P-gp-transfected cells to vinblastine and colchicine and inhibited P-gp's verapamil-stimulated ATPase activity. Half-maximal inhibition of ATPase activity occurred at 13.5 microM disulfiram. Disulfiram (at 100 microM) inhibited a P-gp mutant by 43% (95% confidence interval [CI] = 37% 48%) when cysteine was present at position 431 only and by 72% (95% CI = 66%-77%) when cysteine was present at position 1074 only. Treatment of P-gp-transfected cells with 50 nM disulfiram blocked maturation of recombinant P-gp. CONCLUSIONS: Disulfiram can potentially reduce P-gp-mediated drug resistance by inhibiting P gp activity (possibly via cysteine modification) and/or by blocking its maturation. These results suggest that disulfiram has the potential to increase the efficacy of drug therapies for cancer and acquired immunodeficiency syndrome. PMID- 10841825 TI - Combined treatment with buserelin and tamoxifen in premenopausal metastatic breast cancer: a randomized study. AB - BACKGROUND: Surgical or medical castration and antiestrogenic treatment with tamoxifen are common endocrine treatments for premenopausal women with breast cancer. However, tamoxifen therapy induces high levels of plasma estradiol, with unknown long-term effects. In this study, we investigated the effect of combining estrogen suppression with the luteinizing hormone-releasing hormone agonist buserelin and estradiol receptor blockade with tamoxifen to determine whether the high estradiol levels induced by tamoxifen could be reduced and whether the antitumor effects would be better. METHODS: In a three-arm, randomized, prospective trial, from 1988 through 1995, a total of 161 premenopausal patients with advanced breast cancer were randomly assigned to treatment with buserelin, tamoxifen, or both. Patients with steroid receptor-negative tumors or with tumors of unknown receptor status who had a disease-free interval of less than 2 years were excluded. The median follow-up was 7.3 years, during which 76% of the patients died, all of breast cancer. Patient and tumor characteristics were well balanced among treatment groups. All P values are from two-sided tests. RESULTS: Combined treatment with buserelin and tamoxifen was superior to treatment with buserelin or tamoxifen alone by objective response rate (48%, 34%, and 28% of patients who could be evaluated, respectively; P =.11 [chi(2) test]), median progression-free survival (9.7 months, 6.3 months, and 5.6 months; P =.03), and overall survival (3.7 years, 2.5 years, and 2.9 years; P =.01). Actuarial 5-year survival percentages were 34.2% (95% confidence interval [CI] = 20.4%-48.0%), 14.9% (95% CI = 3.9%-25.9%), and 18.4% (95% CI = 7.0%-29.8%), respectively. No differences in antitumor effects were observed between single-agent treatment groups. During combined treatment or treatment with buserelin alone, plasma estradiol levels were suppressed equally; in contrast, during treatment with tamoxifen alone, plasma estradiol levels increased threefold to fourfold over pretreatment levels. CONCLUSION: Combined treatment with buserelin and tamoxifen was more effective and resulted in longer overall survival than treatment with either drug alone. PMID- 10841826 TI - Influence of cellular ganglioside depletion on tumor formation. AB - BACKGROUND: Gangliosides are immunosuppressive cell surface molecules that are often present in high concentrations in and shed actively by tumor cells. These molecules inhibit the antitumor immune response that is implicated in tumor rejection. We therefore determined the ability of tumor cells pharmacologically depleted of gangliosides to form tumors in mice. METHODS: We tested a ganglioside rich subline of B16 murine melanoma, MEB4, and MEB4 cells that had been depleted of endogenous gangliosides by incubation with 0.5 microM 1-phenyl-2 hexadecanoylamino-3-pyrrolidino-1-propanol, a specific inhibitor of the enzyme glucosylceramide synthase. Tumor formation was assessed twice a week for 10 weeks after the intradermal injection of tumor cells, and metastatic potential was assessed 4 weeks after tail vein injection of tumor cells. All P values are from two-sided tests. RESULTS: Reduction of the ganglioside content of MEB4 cells, which was not cytotoxic to cells and did not inhibit cell proliferation in vitro, markedly reduced their ability to form tumors. Only 40% of the mice given an intradermal injection of 10(5) ganglioside-depleted MEB4 cells developed tumors compared with 100% of the mice given an injection of 10(5) control MEB4 cells (P<.001). Ganglioside depletion also reduced metastasis: A mean of five pulmonary metastases was detected per mouse given an injection of 2 x 10(5) ganglioside depleted MEB4 cells compared with a mean of 25 per mouse given an injection of 2 x 10(5) control MEB4 cells. CONCLUSION: Tumor cells with a pharmacologically decreased concentration of gangliosides produce fewer tumors in mice than do untreated cells, suggesting that pharmacologic depletion of gangliosides should be explored further as a therapeutic approach to cancer. PMID- 10841827 TI - Somatic mutation screening: identification of individuals harboring K-ras mutations with the use of plasma DNA. AB - BACKGROUND: Many cancers are attributed to somatic mutation of DNA. We investigated whether it is feasible to detect cancer-associated somatic mutations in patients with neoplasms by using plasma DNA. METHODS: Plasma samples were prospectively collected from 240 patients undergoing colonoscopy. Colorectal biopsies were performed as clinically indicated in 135 patients, and risk factor information was available from 232 patients. DNA was extracted from plasma and colorectal tissue and was amplified by use of a polymerase chain reaction method that enriches for mutations in codon 12 of the K-ras oncogene. Molecular, histologic, and clinical data were compared by use of two-sided Fisher's exact test. RESULTS: Mutations in the K-ras gene detected in the plasma of 64 (28%) of 232 patients were statistically significantly associated with colorectal cancer risk factors (P =.0002). Of those patients having tissue available for comparison (n = 135), mutations in the K-ras gene were found in the tissues of 35 patients, and 29 (83%) of these 35 showed mutations in plasma samples. In contrast, the plasma assay was negative in 93 of the 100 patients whose tissue K-ras was wild type. Among patients without biopsies (n = 105), 28 had mutated K-ras in their plasma DNA, despite the absence of remarkable colonoscopy findings; 24 of these 28 patients had risk factors for colorectal cancer. Overall, 25 (39%) of 64 patients showing mutations in plasma DNA had colorectal neoplasms with K-ras mutations compared with five (3%) of 176 patients without K-ras mutations in plasma DNA. CONCLUSION: Plasma DNA assays for the detection of mutations in K-ras codon 12 may provide a feasible method to screen populations for somatic mutations frequently found in neoplasms. The clinical utility of using this test in screening populations requires further study. PMID- 10841828 TI - Altered PTEN expression as a diagnostic marker for the earliest endometrial precancers. AB - BACKGROUND: PTEN tumor suppressor gene mutations are the most frequent genetic lesions in endometrial adenocarcinomas of the endometrioid subtype. Testing the hypothesis that altered PTEN function precedes the appearance of endometrial adenocarcinoma has been difficult, however, partly because of uncertainties in precancer diagnosis. METHODS: Two series of endometrial cancer and precancer (endometrial intraepithelial neoplasia, as diagnosed by computerized morphometric analysis) tissue samples were studied, one for PTEN mutations by the use of denaturing gradient gel electrophoresis and another for PTEN protein expression by immunohistochemistry. Endometria altered by high estrogen levels that are unopposed by progestins-conditions known to increase cancer risk-were also studied by immunohistochemistry. Fisher's exact test was used for statistical analysis. RESULTS: The PTEN mutation rate was 83% (25 of 30) in endometrioid endometrial adenocarcinomas and 55% (16 of 29) in precancers, and the difference in number of mutations was statistically significant (two-sided P =.025). No normal endometria showed PTEN mutations. Although most precancers and cancers had a mutation in only one PTEN allele, endometrioid endometrial adenocarcinomas showed complete loss of PTEN protein expression in 61% (20 of 33) of cases, and 97% (32 of 33) showed at least some diminution in expression. Cancers and most precancers exhibited contiguous groups of PTEN-negative glands, while endometria altered by unopposed estrogens showed isolated PTEN-negative glands. CONCLUSIONS: Loss of PTEN function by mutational or other mechanisms is an early event in endometrial tumorigenesis that may occur in response to known endocrine risk factors and offers an informative immunohistochemical biomarker for premalignant disease. Individual PTEN-negative glands in estrogen-exposed endometria are the earliest recognizable stage of endometrial carcinogenesis. Proliferation into dense clusters that form discrete premalignant lesions follows. PMID- 10841829 TI - Immunomodulatory gene therapy with interleukin 12 and 4-1BB ligand: long- term remission of liver metastases in a mouse model. AB - BACKGROUND: The success of immunomodulatory cancer therapy is frequently hampered by the transient nature of the antitumor immune response. We have shown previously in a mouse model that interleukin 12 (IL-12) generates a strong natural killer (NK) cell-mediated antitumor response and reduces liver metastases induced by a colon carcinoma cell line. However, only a small percentage of the treated animals developed the cytotoxic T-lymphocytic response required for a long-term systemic antitumor immunity. 4-1BB is a co-stimulatory molecule expressed on the surface of activated T cells. Interaction of 4-1BB with its natural ligand (4-1BBL) has been shown to amplify T-cell (especially CD8+) mediated immunity. In this study, we investigated the effects of adenovirus mediated gene therapy delivering both IL-12 and 4-1BBL genes on mice with hepatic metastases induced by colon cancer cells. METHODS: Syngeneic BALB/c mice received intrahepatic injection of poorly immunogenic MCA26 colon cancer cells. Various combinations of replication-defective adenoviruses expressing IL-12 and 4-1BBL genes were injected into the established liver tumors. Changes in tumor size and animal survival were then monitored. All statistical tests were two-sided. RESULTS: The long-term survival rate of mice treated with the combination of IL 12 and 4-1BBL was significantly improved over that of animals in the control group (P =.0001). In vivo depletion of NK cells or CD8+ T cells completely abolished the long-term survival advantage of the IL-12 plus 4-1BBL-treated animals (P<.002). Moreover, the systemic immunity induced by this combination treatment protected these animals against a subcutaneous challenge with parental MCA26 cells. CONCLUSION: Adenovirus-mediated transfer of IL-12 and 4-1BBL genes directly into liver tumors resulted in tumor regression that required both NK and CD8+ T cells and generated a potent, long-lasting antitumor immunity. PMID- 10841830 TI - Tissue microarray assessment of prostate cancer tumor proliferation in African- American and white men. PMID- 10841831 TI - The feisty Woman's breast cancer book PMID- 10841833 TI - Re: History of breast-feeding in relation to breast cancer risk: a review of the epidemiologic literature. PMID- 10841834 TI - Re: History of breast-feeding in relation to breast cancer risk: a review of the epidemiologic literature. PMID- 10841835 TI - Response to corrrespondence by nelson PMID- 10841837 TI - RESPONSE: re: estimation of Tamoxifen's efficacy for preventing the formation and growth of breast tumors PMID- 10841836 TI - Re: Estimation of tamoxifen's efficacy for preventing the formation and growth of breast tumors. PMID- 10841838 TI - Saved by the bell: the role of telephone helpline services in the context of mass media anti-smoking campaigns. PMID- 10841839 TI - Youth access: the baby and the bath water. PMID- 10841840 TI - Thailand: winning battles, but the war's far from over. PMID- 10841841 TI - Australia: smoking "K.s" PMID- 10841842 TI - Uganda: athletes fight BAT's abuse of sports. PMID- 10841843 TI - Bangladesh: voyage of disdain sunk without trace. PMID- 10841844 TI - USA: the art of simple dying. PMID- 10841846 TI - Kazakhstan: PM's "PR department" ignores tobacco PMID- 10841847 TI - Tobacco's gulf war against health. PMID- 10841845 TI - Japan: streets unsafe as machines prey on children. PMID- 10841848 TI - The big disappointment: USA weak on convention. PMID- 10841849 TI - Targeting youth and concerned smokers: evidence from Canadian tobacco industry documents. AB - OBJECTIVE: To provide an understanding of the targeting strategies of cigarette marketing, and the functions and importance of the advertising images chosen. METHODS: Analysis of historical corporate documents produced by affiliates of British American Tobacco (BAT) and RJ Reynolds (RJR) in Canadian litigation challenging tobacco advertising regulation, the Tobacco Products Control Act (1987): Imperial Tobacco Limitee & RJR-Macdonald Inc c. Le Procurer General du Canada. RESULTS: Careful and extensive research has been employed in all stages of the process of conceiving, developing, refining, and deploying cigarette advertising. Two segments commanding much management attention are "starters" and "concerned smokers". To recruit starters, brand images communicate independence, freedom and (sometimes) peer acceptance. These advertising images portray smokers as attractive and autonomous, accepted and admired, athletic and at home in nature. For "lighter" brands reassuring health concerned smokers, lest they quit, advertisements provide imagery conveying a sense of well being, harmony with nature, and a consumer's self image as intelligent. CONCLUSIONS: The industry's steadfast assertions that its advertising influences only brand loyalty and switching in both its intent and effect is directly contradicted by their internal documents and proven false. So too is the justification of cigarette advertising as a medium creating better informed consumers, since visual imagery, not information, is the means of advertising influence. PMID- 10841850 TI - Impact of a telephone helpline for smokers who called during a mass media campaign. AB - OBJECTIVE: To evaluate the impact of a telephone helpline (Quitline) with additional support (written information) on callers who use the service during a mass media campaign. DESIGN: Telephone recall surveys of callers to the helpline carried out two months and one year after their initial call. SETTING: Telephone helpline. SUBJECTS: Callers to the helpline. MAIN OUTCOME MEASURES: Smoking behaviour change among callers to the helpline at two months and one year. RESULTS: At one year 22% (95% confidence interval (CI) 18.4% to 25.6%) of smokers reported that they had stopped smoking. Assuming that those who refuse to take part in the one year follow up are continuing smokers and a further 20% of reported successes fail biochemical validation, this yields an adjusted quit rate of 15. 6% (95% CI 12.7% to 18.9%) at one year. Among ex-smokers, 41% (95% CI 34.3% to 47.7%) reported that they were still not smoking at one year. The adjusted figure for ex-smokers at one year is 29% (95% CI 23.3% to 34.8%). Of those who resumed smoking 28% were smoking less than they had been initially. Currently Quitline receives around half a million calls in the course of one year, 93% of whom are phoning for themselves. This represents 4.2% of the total population of adults smokers in England. CONCLUSION: The Health Education Authority's advertising campaign was extremely successful in generating calls to the helpline. Very large numbers of smokers from diverse backgrounds, including the key groups highlighted in the UK government's recent proposals on tobacco, called the Quitline, which appeared to be very successful in helping these callers to stop smoking. For a single intervention to reach 4.2% of the total population of adult smokers in England is a major achievement. This makes Quitline a very promising model for public health intervention programs. PMID- 10841851 TI - New Zealand's tobacco control programme 1985-1998. AB - OBJECTIVE: To review the impact of New Zealand's tobacco control programme from 1985 to 1998 on smoking prevalence and tobacco consumption, and to estimate the scope for further reduction. DESIGN: Country case study; interventions, with outcomes ranked internationally across time. SETTING: New Zealand 1985-98; for 1985-95, 23 OECD countries. INTERVENTIONS: Between 1985 and 1998, New Zealand eliminated tobacco advertising, halved the affordability of cigarettes, and reduced smoke exposure in work time by 39%. MAIN OUTCOME MEASURE: Reduction in adult smoking prevalence and in tobacco products consumption per adult. RESULTS: Changes in prevalence 1985-98: in adults (aged 15+ years), -17% (from 30% to 25%) but short of the 20% target for 2000; in youth (aged 15-24 years), -20% (from 35% to 28%); and in Maori adults (aged 15+ years), -17% (from 56% in 1981 to 46% in 1996). Changes in consumption 1985-98: tobacco products per adult aged 15+ years, -45% (2493 to 1377 cigarette equivalents); cigarettes smoked per smoker, -34% (22. 7 to 15.0 per day). Between 1985 and 1995 New Zealand reduced tobacco products consumption per adult more rapidly than any other OECD country, and reduced youth prevalence more rapidly than most. The acceleration of the decline in cigarette attributable mortality rates in men and in women age 35-69 years averted an additional 1400 deaths between 1985 and 1996. Between 1981 and 1996 smoking prevalence among blue collar workers decreased only marginally, and in 14 15 year olds, rose by one third between 1992 and 1997. CONCLUSION: In 13 years, New Zealand's tobacco control programme has been successful in almost halving tobacco products consumption, particularly by lowering consumption per smoker. With strong political support for quit campaigns, increased taxation, and the elimination of displays of tobacco products on sale, the consumption could theoretically be halved again in as little as 3-6 years. PMID- 10841852 TI - Exposure to cigarette promotions and smoking uptake in adolescents: evidence of a dose-response relation. AB - OBJECTIVE: To assess whether a dose-response relation exists between the number of cigarette promotional items (CPIs) owned by an adolescent, and smoking behaviour. DESIGN AND SETTING: Voluntary, self administered survey of 1265 sixth through to 12th grade students (ages 10-19 years), representing 79-95% of all students attending five rural New Hampshire and Vermont public (state funded) schools in October 1996. The association between the number of CPIs owned by students and smoking behaviour was examined using multivariate regression methods. OUTCOME MEASURES: Adjusted odds of being a smoker (>/= 100 cigarettes lifetime) and, among never and experimental smokers, adjusted cumulative odds of having higher levels on a smoking uptake index given the number of CPIs owned. RESULTS: One third of students owned a CPI (n = 406). Among owners, 211 owned one, 82 owned two, 57 owned three, 24 owned four, 23 owned five, and 7 students owned six CPIs. The number of CPIs owned by students was not associated with grade in school but was significantly higher in males, those with poorer school performance, those who perceived high prevalence of peer smoking, and those with higher exposure to peer and family smoking. The more items a student owned, the greater the chances of being a smoker. For example, smoking prevalence was 11.2% for those not owning a CPI, 41.5% for those owning two, 58.5% for those owning four, and 71.4% for those owning six CPIs. The dose-response relation remained after controlling for confounding; compared with those who did not own a CPI, the likelihood of being a smoker was significantly higher for those who owned one CPI, with an adjusted odds ratio (OR) of 2.7 (95% confidence interval (CI) 1.7 to 4.1); OR was 3.4 (95% CI 1.9 to 5.9) for those owning two CPIs, and 8.4 (95% CI 5.0 to 14.2) for those owning three or more CPIs. After excluding smokers, there was a crude dose-response association between CPI ownership and higher rates of experimentation with cigarettes among sixth to ninth graders (ages 11-15 years) only (n = 543). After controlling for confounding influences, the dose-response relation remained, with the likelihood of being higher on the smoking uptake index rising with the number of CPIs owned: one CPI, adjusted cumulative OR 1.7 (95% CI 1.1 to 2.60); two CPIs, OR 2.5 (95% CI 1.2 to 5.1); and three or more CPIs, OR 4.8 (95% CI 1.9 to 12.2). CONCLUSIONS: This study offers evidence of a dose-response relation between the number of CPIs owned by adolescents and higher likelihood of experimental and established smoking. The dose-response relation persists after controlling for confounding influences. These data provide further support of a causal relation between tobacco promotional campaigns and smoking behaviour among adolescents. PMID- 10841854 TI - Effectiveness of comprehensive tobacco control programmes in reducing teenage smoking in the USA. AB - OBJECTIVE: To describe the extent to which comprehensive statewide tobacco control programmes in the USA have made progress toward reducing teenage smoking. DATA SOURCES: Literature search of Medline for reviews of effectiveness of programme and policy elements, plus journal articles and personal request for copies of publicly released reports and working papers from evaluation staff in each of the state programmes of California, Massachusetts, Arizona, Oregon, and Florida. STUDY SELECTION: All studies, reports, and commentaries that provided information on aspects of programme implementation and evaluation. DATA SYNTHESIS: Statewide comprehensive programmes show high levels of advertising recall and generally positive improvement in smoking related beliefs and attitudes among teenagers. More fully funded programmes lead to increased mass media campaign advertising and community initiatives; a greater capacity to implement school based smoking prevention programmes; and an increase in the passage of local ordinances that create smoke free indoor environments and reduce cigarette sales to youth. The combination of programme activity and increased tobacco tax reduce cigarette consumption more than expected as a result of price increases alone, and these effects seem to apply to adolescents as well as adults. Programmes are associated with a decline in adult smoking prevalence, with these effects observed to date in California, Massachusetts, and Oregon. Arizona and Florida have yet to examine change in adult prevalence associated with programme exposure. California and Massachusetts have demonstrated relative beneficial effects in teenage smoking prevalence, and Florida has reported promising indications of reduced prevalence. Arizona has yet to report follow up data, and Oregon has found no change in teenage smoking, but has only two years of follow up available. One of the most critical factors in programme success is the extent of programme funding, and consequent level of programme implementation, and the degree to which this is undermined by the tobacco industry and other competitors for funding. CONCLUSIONS: Despite the different strengths and combinations of programme messages and strategies used in these comprehensive programmes, there is evidence that they lead to change in factors that influence teenage smoking, and to reductions in teenage smoking. PMID- 10841853 TI - A systematic review of interventions for preventing tobacco sales to minors. AB - OBJECTIVE: To assess the effectiveness of interventions to reduce underage access to tobacco by deterring shopkeepers from making illegal sales. METHOD: Systematic literature review. DATA SOURCES: The Cochrane Tobacco Addiction group specialised register and Medline. Studies of interventions to alter retailer behaviour were identified. The terms used for searching combined terms for smoking and tobacco use with terms for minors, children or young people, and retailers, sales or commerce. STUDY SELECTION: Studies in which there was an intervention with retailers of tobacco, either through education about, or enforcement of, local ordinances. The outcomes were changes in retailer compliance with legislation (assessed by test purchasing), changes in young people's perceived ease of access to tobacco products, and changes in smoking behaviour. Controlled studies with or without random allocation of retail outlets or communities, and uncontrolled studies with pre- and post intervention assessment, were included. DATA EXTRACTION: Two reviewers assessed studies for inclusion. One extracted data with checking by the second. DATA SYNTHESIS: The results were synthesised qualitatively, with greater weight given to controlled studies. Thirteen of 27 included studies used controls. RESULTS: Giving retailers information was less effective in reducing illegal sales than active enforcement and/or multicomponent educational strategies. No strategy achieved complete, sustained compliance. In three controlled trials, there was little effect of intervention on youth perceptions of access or prevalence of smoking. CONCLUSIONS: Interventions with retailers can lead to large decreases in the number of outlets selling tobacco to youths. However, few of the communities studied in this review achieved sustained levels of high compliance. This may explain why there is limited evidence for an effect of intervention on youth perception of ease of access to tobacco, and on smoking behaviour. PMID- 10841856 TI - Effects of market liberalisation on smoking in Japan. AB - OBJECTIVE: To document the effect of the liberalisation of the Japanese tobacco market on Japanese smoking rates and on Japanese tobacco industry practices. DATA SOURCE: Asahi Shimbun (major daily newspaper) from 1980 to 1996. STUDY SELECTION: Review of media coverage on the effects of market liberalisation following the imposition of the USA's section 301 trade sanction. DATA SYNTHESIS: The opening of Japan's tobacco market to foreign cigarette companies stalled a decline in smoking prevalence. Smoking rates among young women increased significantly, and also appear to be on the rise among adolescents. Aggressive marketing and promotional activities by US and Japanese tobacco companies in response to trade liberalisation appear responsible for these adverse trends. Steep increases in sales through vending machines were also possible contributors to the rising smoking prevalence among adolescents. On the positive side, market liberalisation indirectly promoted smoking control efforts in Japan, by causing an anti-smoking movement to coalesce. CONCLUSION: Market liberalisation in Japan played a significant role in increasing smoking prevalence among young women and adolescents while helping to transform the issue of smoking in Japan from a matter of individual choice to a public health problem. PMID- 10841855 TI - Costs of employee smoking in the workplace in Scotland. AB - BACKGROUND: Employers have responded to new regulations on the effects of passive smoking by introducing a range of workplace policies. Few policies include provision of smoking cessation intervention. OBJECTIVE: To estimate the cost to employers of smoking in the workplace in Scotland to illustrate the potential gains from smoking cessation provision. Costs vary with type of smoking policy in place; therefore, to estimate these costs results from a survey were combined with evidence drawn from a literature review. STUDY DESIGN: A telephone survey of 200 Scottish workplaces, based on a stratified random sample of workplaces with 50 or more employees, was conducted in 1996. Additional evidence was compiled from a review of the literature of smoking related costs and specific smoking related effects. RESULTS: 167 completed responses were received, of which 156 employers (93%) operated a smoking policy, 57 (34%) operated smoke free buildings, and 89 (53%) restricted smoking to a "smoke room". The research literature shows absenteeism to be higher among smokers when compared to non smokers. The estimated cost of smoking related absence in Scotland is pound40 million per annum. Total productivity losses are estimated at approximately pound450 million per annum. In addition, the resource cost in terms of losses from fires caused by smoking materials is estimated at approximately pound4 million per annum. In addition, there are costs from smoking related deaths and smoking related damage to premises. CONCLUSION: This study shows how smoking cessation interventions in the workplace can yield positive cost savings for employers, resulting in gains in productivity and workplace attendance which may outweigh the cost of any smoking cessation programme. PMID- 10841857 TI - Smoke knows no boundaries: legal strategies for environmental tobacco smoke incursions into the home within multi-unit residential dwellings. AB - OBJECTIVE: To describe legal theories that non-smoking residents of multiple occupancy buildings may employ when affected by environmental tobacco smoke (ETS) from neighbouring units. DESIGN: Legal research was conducted in several US states. Research was performed among statutes and regulations. State health regulations were examined as well as common law claims of nuisance, warranties of habitability, and the right of quiet enjoyment. RESULTS: Through the use of state regulations, such as a sanitary code, several states provide general language for protecting the health of residents in multi-unit buildings. State law also supports more traditional claims of nuisance, warranties of habitability, and the right of quiet enjoyment. CONCLUSIONS: The use of state regulations has the potential to provide an effective, existing vehicle for resolution of ETS incursion problems. The general health protection language of the regulations, in conjunction with the latest evidence of the harmful effects of ETS, gives state agencies authority to regulate environmental tobacco smoke incursions among apartments in multi-unit dwellings. Where state regulations are not available, other common law legal remedies may be available. PMID- 10841858 TI - Globalisation of tobacco industry influence and new global responses. AB - The globalisation of tobacco marketing, trade, research, and industry influence represents a major threat to public health worldwide. Drawing upon tobacco industry strategy documents prepared over several decades, this paper will demonstrate how the tobacco industry operates as a global force, regarding the world as its operating market by planning, developing, and marketing its products on a global scale. The industry has used a wide range of methods to buy influence and power, and penetrate markets across the world. It has an annual turnover of almost US$400 billion. In contrast, until recently tobacco control lacked global leadership and strategic direction and had been severely underfunded. As part of moving towards a more sustainable form of globalisation, a global enabling environment linked to local actions should focus on the following strategies: global information management; development of nationally and locally grounded action; global regulation, legal instruments, and foreign policy; and establishment of strong partnerships with purpose. As the vector of the tobacco epidemic, the tobacco industry's actions fall far outside of the boundaries of global corporate responsibility. Therefore, global and local actions should not provide the tobacco industry with the two things that it needs to ensure its long term profitability: respectability and predictability. PMID- 10841859 TI - Research priorities for tobacco control in developing countries: a regional approach to a global consultative process. AB - OBJECTIVE: To develop regional tobacco control research agendas for developing countries through a consultative process. METHODS: Research for International Tobacco Control, located at the International Development Research Centre in Ottawa, Canada, convened three regional meetings for Latin America and the Caribbean, South and Southeast Asia, and Eastern, Central and Southern Africa. Participation by researchers, policymakers, and advocates from a wide range of disciplines ensured an accurate representation of regional issues. RESULTS: The four main recurring themes within each regional agenda were: (1) the lack of standardised and comparable data; (2) the absence of a network for communication of information, data, and best practices; (3) a lack of adequate capacity for tobacco control research, especially in non-health related areas such as economics and policy analysis; and (4) a need for concerted mobilisation of human and financial resources in order to implement a comprehensive research agenda, build partnerships, and stimulate comparative research and analysis. Specific research issues included the need for descriptive data with respect to the supply side of the tobacco equation, and analytical data related to tobacco use, production and marketing, and taxation. CONCLUSIONS: There was a uniform perception of tobacco as a multidisciplinary issue. All regional agendas included a balance of health, economic, agricultural, environmental, sociocultural, and international trade concerns. Research data are urgently required to provide a sound basis for the development of tobacco control policies and programmes. As tobacco control takes its rightful place on the global health agenda, it is vital that funding for tobacco control research be increased. PMID- 10841860 TI - Advancing knowledge on regulating tobacco products, Oslo, Norway. PMID- 10841861 TI - World's best practice in tobacco control. PMID- 10841862 TI - What is the future for the tobacco industry? PMID- 10841863 TI - Comprehensive tobacco control strategy needed. PMID- 10841864 TI - Recreational nicotine: uncertain benefits and several major risks [commment]. PMID- 10841866 TI - Shameful science: four decades of the German tobacco industry's hidden research on smoking and health. PMID- 10841865 TI - Compete with the tobacco industry. PMID- 10841867 TI - 147,000 pieces of silver. PMID- 10841868 TI - The centenary of the enactment of the law for prohibiting minors from smoking in Japan. PMID- 10841869 TI - Why joe camel is still smiling PMID- 10841870 TI - Urothelial carcinoma associated with the use of a Chinese herb (Aristolochia fangchi) AB - BACKGROUND: Chinese-herb nephropathy is a progressive form of renal fibrosis that develops in some patients who take weight-reducing pills containing Chinese herbs. Because of a manufacturing error, one of the herbs in these pills (Stephania tetrandra) was inadvertently replaced by Aristolochia fangchi, which is nephrotoxic and carcinogenic. METHODS: The diagnosis of a neoplastic lesion in the native urinary tract of a renal-transplant recipient who had Chinese-herb nephropathy prompted us to propose regular cystoscopic examinations and the prophylactic removal of the native kidneys and ureters in all our patients with end-stage Chinese-herb nephropathy who were being treated with either transplantation or dialysis. Surgical specimens were examined histologically and analyzed for the presence of DNA adducts formed by aristolochic acid. All prescriptions written for Chinese-herb weight-reducing compounds during the period of exposure (1990 to 1992) in these patients were obtained, and the cumulative doses were calculated. RESULTS: Among 39 patients who agreed to undergo prophylactic surgery, there were 18 cases of urothelial carcinoma (prevalence, 46 percent; 95 percent confidence interval, 29 to 62 percent): 17 cases of carcinoma of the ureter, renal pelvis, or both and 1 papillary bladder tumor. Nineteen of the remaining patients had mild-to-moderate urothelial dysplasia, and two had normal urothelium. All tissue samples analyzed contained aristolochic acid-related DNA adducts. The cumulative dose of aristolochia was a significant risk factor for urothelial carcinoma, with total doses of more than 200 g associated with a higher risk of urothelial carcinoma. CONCLUSIONS: The prevalence of urothelial carcinoma among patients with end-stage Chinese-herb nephropathy (caused by aristolochia species) is a high. PMID- 10841871 TI - The causes and risk of stroke in patients with asymptomatic internal-carotid artery stenosis. North American Symptomatic Carotid Endarterectomy Trial Collaborators. AB - BACKGROUND: The causes of stroke in patients with asymptomatic carotid-artery stenosis have not been carefully studied. Information about causes might influence decisions about the use of carotid endarterectomy in such patients. METHODS: We studied patients with unilateral symptomatic carotid-artery stenosis and asymptomatic contralateral stenosis from 1988 to 1997. The causes, severity, risk, and predictors of stroke in the territory of the asymptomatic artery were examined and quantified. RESULTS: The risk of stroke at five years after study entry in a total of 1820 patients increased with the severity of stenosis. Among 1604 patients with stenosis of less than 60 percent of the luminal diameter, the risk of a first stroke was 8.0 percent (1.6 percent annually), as compared with 16.2 percent (3.2 percent annually) among 216 patients with 60 to 99 percent stenosis. In the group with 60 to 99 percent stenosis, the five-year risk of stroke in the territory of a large artery was 9.9 percent, that of lacunar stroke was 6.0 percent, and that of cardioembolic stroke 2.1 percent. Some patients had more than one stroke of more than one cause. In the territory of an asymptomatic occluded artery (as was identified in 86 patients), the annualized risk of stroke was 1.9 percent. Strokes with different causes had different risk factors. The risk factors for large-artery stroke were silent brain infarction, a history of diabetes, and a higher degree of stenosis; for cardioembolic stroke, a history of myocardial infarction or angina and hypertension; for lacunar stroke, age of 75 years or older, hypertension, diabetes, and a higher degree of stenosis. CONCLUSIONS: The risk of stroke among patients with asymptomatic carotid-artery stenosis is relatively low. Forty-five percent of strokes in patients with asymptomatic stenosis of 60 to 99 percent are attributable to lacunes or cardioembolism. These observations have implications for the use of endarterectomy in asymptomatic patients. Without analysis of the risk of stroke according to cause, the absolute benefit associated with endarterectomy may be overestimated. PMID- 10841872 TI - A comparison of paracentesis and transjugular intrahepatic portosystemic shunting in patients with ascites. AB - BACKGROUND: In patients with cirrhosis and ascites, creation of a transjugular intrahepatic portosystemic shunt may reduce the ascites and improve renal function. However, the benefit of this procedure as compared with that of large volume paracentesis is uncertain. METHODS: We randomly assigned 60 patients with cirrhosis and refractory or recurrent ascites (Child-Pugh class B in 42 patients and class C in 18 patients) to treatment with a transjugular shunt (29 patients) or large-volume paracentesis (31 patients). The mean (+/-SD) duration of follow up was 45+/-16 months among those assigned to shunting and 44+/-18 months among those assigned to paracentesis. The primary outcome was survival without liver transplantation. RESULTS: Among the patients in the shunt group, 15 died and 1 underwent liver transplantation during the study period, as compared with 23 patients and 2 patients, respectively, in the paracentesis group. The probability of survival without liver transplantation was 69 percent at one year and 58 percent at two years in the shunt group, as compared with 52 percent and 32 percent in the paracentesis group (P=0.11 for the overall comparison, by the log rank test). In a multivariate analysis, treatment with transjugular shunting was independently associated with survival without the need for transplantation (P=0.02). At three months, 61 percent of the patients in the shunt group and 18 percent of those in the paracentesis group had no ascites (P=0.006). The frequency of hepatic encephalopathy was similar in the two groups. Of the patients assigned to paracentesis in whom this procedure was unsuccessful, 10 received a transjugular shunt a mean of 5.5+/-4 months after randomization; 4 had a response to this rescue treatment. CONCLUSIONS: In comparison with large-volume paracentesis, the creation of a transjugular intrahepatic portosystemic shunt can improve the chance of survival without liver transplantation in patients with refractory or recurrent ascites. PMID- 10841873 TI - Long-term follow-up of unilateral pallidotomy in advanced Parkinson's disease. AB - BACKGROUND: Although the short-term benefits of posteroventral pallidotomy for patients with advanced Parkinson's disease have been well documented, little is known about the long-term outcome of the procedure. METHODS: We conducted a long term follow-up study of a cohort of 40 patients who had undergone unilateral posteroventral medial pallidotomy between 1993 and 1996. Twenty patients were not evaluated because they had undergone a second surgical procedure (11 patients) or had died (2) or because they had dementia or another debilitating illness (4), lived too far away (1), or had been lost to follow-up (2). We conducted serial postoperative assessments of parkinsonism in the remaining 20 patients while they were taking medications ("on" period) and after overnight withdrawal of the drugs ("off" period). The mean follow-up time was 52 months (range, 41 to 64). RESULTS: The combined off-period score for activities of daily living and motor function on the Unified Parkinson's Disease Rating Scale was 18.0 percent better at the last evaluation than at base line (95 percent confidence interval, 4.9 to 31.0 percent; P=0.01). Significant improvements were also evident in the off-period scores for contralateral tremor (65.4 percent improvement, P=0.007), rigidity (43.2 percent, P=0.03), and bradykinesia (18.2 percent, P=0.04) and in the on period score for contralateral dyskinesia (70.6 percent, P<0.001). Changes in medication did not contribute to the sustained improvement. The 20 patients who could not be included in the long-term analysis had similar base-line characteristics but a worse response to surgery at six months. CONCLUSIONS: In the group of patients with advanced Parkinson's disease who could be enrolled in our long-term follow-up study of unilateral posteroventral medial pallidotomy (20 patients from the original cohort of 40), significant early improvements in off period contralateral signs of parkinsonism were sustained for up to five and a half years. There was a sustained significant improvement in on-period contralateral dyskinesia but not in other on-period signs of parkinsonism. PMID- 10841874 TI - Images in clinical medicine. Severe malaria. PMID- 10841875 TI - Health advice and immunizations for travelers. PMID- 10841876 TI - Pediatric surgery. Second of two parts. PMID- 10841877 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 17-2000. A 21-week-old girl with fever and a renal mass. PMID- 10841878 TI - Cancer and herbs. PMID- 10841879 TI - Carotid endarterectomy revisited. PMID- 10841880 TI - The role of transjugular portosystemic shunting in patients with ascites. PMID- 10841881 TI - Ethical considerations in the treatment of infertility in women with human immunodeficiency virus infection. PMID- 10841882 TI - Nitric oxide (NO.) stabilizes whereas nitrosonium (NO+) enhances filopodial outgrowth by rat retinal ganglion cells in vitro. AB - Recent observations suggest that nitric oxide (NO(.)) can increase or decrease growth cone motility. Here, these apparently paradoxical results are explained by distinct actions of different NO-related species. Filopodial morphology of 223 rat retinal ganglion cells was monitored under computer-enhanced video microscopy in the presence of NO synthase (NOS) substrates or inhibitors, donors of specific NO-related species, and membrane-permeant cyclic nucleotide analogs. Physiological NOS activity induced filopodial outgrowth, whereas inhibition of NOS stabilized filopodia. Similar to NOS, nitrosonium (NO(+) transfer) and peroxynitrite (ONOO(-)), which can regulate the activity of growth-associated proteins by S-nitrosylation and oxidation, respectively, induced filopodial outgrowth. In contrast, NO(.), which stimulates guanylate cyclase to increase cGMP, stabilized filopodial activity. Thus disparate NO-related species may offer a dynamic process of filopodial growth regulation. PMID- 10841883 TI - A comparison of strain-related susceptibility in two murine recovery models of global cerebral ischemia. AB - Genetically engineered mice are increasingly important in stroke research. The strains on which these constructs are built are known to have inherent differential sensitivities to ischemic insults. This has been largely attributed to differences in vascular anatomy. This study compared the outcome from forebrain ischemia in two common murine background strains using two different types of ischemic insult. C57Bl/6 and SV129 mice were subjected to two vessel (bilateral carotid) occlusion (2VO) or 2VO plus systemic hypotension (2VO+Hypo; mean arterial pressure=30+/-2 mmHg) for 10-20 min. Ventilation and pericranial temperature were controlled. Cerebral blood flow (CBF) was determined by 14C iodoantipyrine autoradiography. Histologic damage in forebrain structures was measured 3 days post-ischemia. During 2VO+Hypo, the EEG became isoelectric in all animals. During 2VO alone, EEG isoelectricity occurred in 73% of C57Bl/6 and 50% of SV129 mice. Forebrain CBF was reduced to a similar extent in both strains. Greater CBF variability was seen with 2VO alone versus 2VO+Hypo. CBF was less in the 2VO+Hypo model. SV129 mice had wider posterior communicating but smaller basilar artery diameters. With or without hypotension, SV129 mice had markedly less severe histologic damage than C57Bl/6 mice. A time-dependent increase in histologic damage was demonstrated in the 2VO+Hypo model but not with 2VO alone. The 2VO and 2VO+Hypo models produced similar magnitudes of histologic injury in C57Bl/6 mice subjected to 10-min ischemia. SV129 mice were resistant to ischemia in either model. The 2VO+Hypo model produced a more uniform severity of ischemia as defined by CBF and EEG examination. Despite this, the murine strain had a substantially greater impact on histologic outcome than did cerebrovascular anatomy or the type of model used to produce the ischemic insult. PMID- 10841884 TI - Localization of thyroid hormone receptor beta2 in the ventral medullary neurons that synthesize thyrotropin-releasing hormone. AB - Altered thyroid statuses are associated with autonomic disorders. Thyrotropin releasing hormone (TRH) synthesized in medullary raphe pallidus (Rpa), raphe obscurus (Rob) and the parapyramidal regions (PPR) regulates vagal and sympathetic preganglionic motoneurons. Hypothyroidism increased TRH gene expression and c-Fos immunoreactivity (IR) in these nuclei. Whether these increases represent a direct action of thyroid hormone was studied by detecting the presence of thyroid hormone receptor beta2 (TRbeta2) in pro-TRH-synthesizing neurons in the Rpa, Rob and the PPR using immunohistochemistry with specific TRbeta2 antiserum and in situ hybridization with digoxigenin-labeled pro-TRH cRNA probe. TRbeta2 IR was widely distributed throughout the medulla and primarily localized within the cell nuclei. Particularly intense immunostaining was presented in the Rpa, Rob and the PPR neurons. The combination of immunohistochemistry with in situ hybridization revealed that all pro-TRH mRNA positive neurons in these ventral medullary nuclei were also TRbeta2 IR positive. The numbers of TRbeta2 IR-positive neurons in each nucleus were identical in both euthyroid rats and hypothyroid rats induced by 6-n-propyl-2-thiouracil in drinking water for 4 weeks. The finding that TRbeta2 localized in pro-TRH synthesizing neurons in the ventral medullary nuclei provides an anatomical substrate for a direct thyroid hormone action on these neurons in the regulation of TRH gene expression, which may contribute to the altered autonomic activity in different thyroid statuses. PMID- 10841885 TI - CREB contributes to the increased neurite outgrowth of sensory neurons induced by vasoactive intestinal polypeptide and activity-dependent neurotrophic factor. AB - Our recent experiments suggest that vasoactive intestinal polypeptide (VIP) enhances neurite outgrowth of dissociated rat dorsal root ganglion cells, indirectly, via the release of a trophic factor from the spinal cord. In this study, we have examined the possible contribution of activity-dependent neurotrophic factor (ADNF) to the trophic actions of VIP. In addition, as we have shown that the factor mediating the trophic actions of VIP acts via protein kinase A we have also examined the contribution of CREB, which is a transcription factor activated by protein kinase A. As previously shown, supernatant taken from spinal cord incubated with VIP, significantly increased the percentage of sensory neurons with neurites. Antiserum against ADNF attenuated the trophic effect of the VIP-conditioned supernatant. Consistently, the ADNF agonist, ADNF(14) (0.001 0.1 fM), significantly enhanced the percentage of cells with neurite outgrowth. Furthermore, the trophic action of ADNF(14) was attenuated by a protein kinase A inhibitor, Rp-cAMPS, whereas the inactive isomer, Sp-cAMPS, had no effect. Preincubation of cells with 5 mcM CREB antisense oligonucleotides, attenuated the increase in neurite outgrowth induced by either the supernatant or ADNF(14). The sense oligonucleotide had no influence on the enhanced neurite outgrowth. We also found that both the supernatant and ADNF(14) induced an increase in the percentage of cells expressing phosphorylated CREB. The data suggests that VIP induces a release of neurotrophic factors, such as ADNF, which enhance neurite outgrowth. In addition, protein kinase A and CREB appear to contribute to the neurotrophic actions of VIP and ADNF. The mechanisms underlying the neurotrophic action of VIP, may have important implications for sprouting and/or synaptic reorganization of central terminals of sensory neurons, which may contribute to neuropathic pain that commonly occurs following peripheral nerve damage. PMID- 10841886 TI - Differential modulatory effects of norepinephrine on synaptically driven responses of layer V barrel field cortical neurons. AB - The effects of norepinephrine (NE) and the alpha-1 agonist phenylephrine (PE) on synaptically evoked responses of electrophysiologically identified pyramidal neurons in layer V of rat somatosensory cortex were studied in brain slices using intracellular recording techniques. When added to the bathing medium NE (10 microM) tended to increase the synaptic responsiveness of regular spiking neurons and decrease the responsiveness of intrinsic burst neurons. NE had mixed effects on layer V cells which were characterized as intermediate types between regular spiking and intrinsic burst neurons. PE exerted a similar spectrum of actions on layer V cortical neurons. For both adrenergic agents the greatest facilitating effect was observed on responses to low intensity synaptic stimulation. These results suggest that NE exerts different modulatory actions on different electrophysiologically-defined classes of layer V sensory cortical neurons. PMID- 10841887 TI - NOC/oFQ contributes to hypoxic-ischemic impairment of N-methyl-D-aspartate induced cerebral vasodilation. AB - Previous studies in piglets show that either hypoxia, ischemia-reperfusion (I+R) or combined hypoxia-ischemia-reperfusion (H+I+R) attenuated N-methyl-D-aspartate (NMDA)-induced pial artery dilation. This study was designed to determine the contribution of the newly described opioid nociceptin orphanin FQ (NOC/oFQ) to hypoxic-ischemic impairment of NMDA induced cerebral vasodilation in piglets equipped with a closed cranial window. Global cerebral ischemia was produced via elevated intracranial pressure. Hypoxia decreased P(O(2)) to 35+/-3 mmHg with unchanged P(CO(2)). I+R elevated CSF NOC/oFQ from 67+/-4 to 266+/-29 pg/ml ( approximately 10(-10) M) while H+I+R elevated CSF NOC/oFQ to 483+/-67 pg/ml within 1 h of reperfusion. Such elevated NOC/oFQ levels returned to control within 4 h in I+R animals and within 12 h in H+I+R animals. Topical NOC/oFQ (10( 10) M) had no effect on pial artery diameter by itself but attenuated NMDA (10( 8), 10(-6) M) induced pial dilation (control, 9+/-1 and 16+/-1; coadministered NOC/oFQ, 5+/-1 and 10+/-1%). NMDA induced pial artery dilation was attenuated by I+R or H+I+R; but such dilation was partially restored by pretreatment with the putative NOC/oFQ antagonist [F/G] NOC/oFQ (1-13) NH(2) (10(-6) M) (control, 9+/-1 and 16+/-1; I+R, 3+/-1 and 5+/-1; I+R+NOC/oFQ antagonist, 6+/-1 and 11+/-1%) Similar results were obtained for glutamate. These data suggest that NOC/oFQ release contributes to impaired NMDA and glutamate-induced cerebrovasodilation following I+R or H+I+R. PMID- 10841888 TI - Multiple representations of information in the primary auditory cortex of cats. I. Stability and change in slow components of unit activity after conditioning with a click conditioned stimulus. AB - Recordings of activity were made from 647 single units of the A(I) cortex of awake cats to evaluate behavioral state-dependent changes in the population response to a 70-dB click. Averages of PST histograms of unit activity were used to assess the changes in response. This report focuses on slow components of the responses disclosed by averages employing bin widths of 16 ms. Responses were compared before and after a Pavlovian blink CR was produced by forward pairing of click conditioned stimuli (CSs) with USs. A backward-paired 70-dB hiss was presented as a discriminative stimulus. Studies were also done after backward pairing of the click CSs (backward conditioning) that produced weak sensitization instead of a conditioned response. There were four main findings. First, components of activity elicited 32-160 ms after presenting the hiss decreased significantly after conditioning and after backward conditioning. The decreases after conditioning represented the most pronounced changes in activity evoked by either clicks or hisses in this behavioral state. Second, baseline firing decreased after both conditioning and backward conditioning. The direction of baseline change was opposite that found in adjacent cortical regions and in A(I) cortex after operant conditioning employing an acoustic cue. Third, prior to conditioning, unit activity in response to the hiss declined before the sound of the hiss reached its peak or terminated. This decrease was thought to represent a habituatory adaptation of response to a prolonged acoustic stimulus. This type of habituation to a lengthy stimulus has been recognized, behaviorally, but has not been observed previously in the activity of units of the auditory receptive cortex. Fourth, the percentage of click responsive units did not change significantly after the click was used as a CS for conditioning, and despite the accompanying changes in baseline activity, the absolute levels of activity summed in the first 16 ms after click delivery remained stable across behavioral states in which the motor response to the click was altered profoundly. The onset of the conditioned motor response began 20 ms after the click, and was shown earlier to depend on rapid, potentiated transmission through the cochlear nucleus and motor cortex for its generation. Thus the stability of the response to the click in the primary auditory receptive cortex was unexpected. This led us to make further analyses of the data with 2- and 4-ms bin widths (see companion report) that eventually disclosed a potentiated response to the click. The findings show stability and change in the response to the click as a CS, depending on the band pass (bin width) used for analysis of spike activity. In the representation disclosed by low pass filtering in this study, the response was stable. This representation provided information suitable for identifying commonalties of the click signals across varying behavioral states. The representations of the click and hiss contained in the slow components of the population response in the A(I) cortex were uncorrelated with the selective potentiation of activity in motor cortex and behavioral performance in response to click as a CS after conditioning. Although changes in the activity evoked by hisses occurred after conditioning, the changes also occurred after backward conditioning when only small, sensitized behavioral responses to clicks and hisses were observed. Basic theoretical considerations about information transmission in complex neural networks plus clinical observations comparing derangements of linguistic and non linguistic cortical functions in humans suggest that multiple representations of conditioned stimulus inputs may exist in local populations of cortical neurons. Together, our studies provide evidence for two different, concurrent representations of information about a click CS encoded in the spike activity of the A(I) cortex. PMID- 10841889 TI - Multiple representations of information in the primary auditory cortex of cats. II. Stability and change in early (<32 ms), rapid components of activity after conditioning with a click conditioned stimulus. AB - Activity was recorded from single units of the A(I) cortex of awake animals to identify early (<32 ms) components of the population response to a 70 dB click and establish if they changed after using the click as a CS for conditioning. A 70 dB hiss was used as a discriminative stimulus. Responses to these stimuli were compared before and after a forward order of pairing that produced conditioning and a backward order of pairing that produced weak sensitization (backward conditioning). Averages of discharges in 2 and 4 ms bins distinguished primary (8 12 ms) from secondary (12-16 ms) temporal components of response to the click, and confirmed that the onset of the response was shorter in A(I) (8 ms, mean of 647 units) than in the adjacent, A(II) cortex (16 ms, mean of 95 units). (All times include a 1.6 ms transmission delay in sound arrival.) Primary and secondary components of A(I) responses to click did not change uniformly after changes in behavioral state, and were affected differently by both conditioning and backward conditioning. The percentage of cells with onsets of response to the click at secondary latencies (and to the hiss at tertiary latencies) increased after backward conditioning but not after conditioning, as did the magnitude of activity in response to the click. (The latter had a lesser degree of increase after conditioning.) The primary response to the click did not show these increases. The non-uniform changes suggested that temporal processing of the click was conducted differently in the 8-12 ms post stimulus period than in the 12-16 ms period. Within the total population of cells, it was possible to identify a small subgroup (13%) of highly auditory-responsive units that showed an increased primary response to the click as a CS selectively after conditioning and not after backward conditioning. The secondary component of response in these cells increased after both conditioning and backward conditioning. The percentages of cells responding to the click and hiss at primary latencies did not change significantly after conditioning, even in the subgroup of highly responsive cells. The results characterize differently timed components of rapid responses to acoustic stimuli in the A(I) cortex, disclose significant temporal differences in primary, secondary and tertiary information processing that affect the representations of the transmitted acoustic message across different behavioral states, and find one representation in a small subgroup of cells that supports the hypothesis that cells of the A(I) cortex have a selectively potentiated response to the CS after conditioning. PMID- 10841890 TI - Behavioral characterization of neuropeptide Y knockout mice. AB - An extensive behavioral characterization was conducted with mice lacking the gene for neuropeptide Y (NPY) including response to 24 and 48 h fast and challenge with small molecule antagonists of NPY receptors implicated in mediating the feeding effects of NPY (i.e., Y1 and Y5). In addition, wildtype (WT) and NPY knockout (KO) mice were tested in locomotor monitors, elevated plus maze, inhibitory avoidance, acoustic startle, prepulse inhibition, and hot plate assays. One of the major findings was that the NPY KO mice have a reduced food intake relative to WT controls in response to fasting. Also, based on data from the behavioral models, the NPY KO mice may have an anxiogenic-like phenotype, and appear to be hypoalgesic in the hot plate paradigm. The data from these studies provide further evidence of involvement of NPY in energy balance, anxiety, and possibly nociception. PMID- 10841891 TI - Intrastriatal administration of 5-aminolevulinic acid induces convulsions and body asymmetry through glutamatergic mechanisms. AB - The involvement of glutamatergic and GABAergic mechanisms in the behavioral effects induced by the intrastriatal injection of 5-aminolevulinic acid (ALA) (1 8 mgr;mol/2 mgr;l), a metabolite that accumulates in porphyrias, was evaluated. ALA administration to adult female rats increased locomotor activity, induced clonic convulsions and elicited dose-dependent body asymmetry assessed by the elevated body swing test. ALA-induced convulsions were prevented by intrastriatal preadministration of the glutamate antagonists, 6, 7-dinitroquinoxaline-2,3-dione (8 nmol/0.5 microl) or dizocilpine (2. 5 nmol/0.5 microl), but not by the GABA agonist, muscimol (46 pmol/0. 5 microl). Body asymmetry was prevented only by 6, 7-dinitroquinoxaline-2,3-dione pretreatment. A higher dose of muscimol (92 pmol/0.5 microl) prevented both ALA-induced convulsions and body asymmetry. However, this dose of muscimol induced motor biases, which make difficult to ascertain the involvement of GABA(A) receptors in ALA-induced behavioral effects. This study suggests that glutamatergic mechanisms underlie the ALA-induced convulsions and body asymmetry. The present results may be of value in understanding the physiopathology of the neurological dysfunction occurring in acute porphyrias. PMID- 10841892 TI - Estrogen reduces acute striatal dopamine responses in vivo to the neurotoxin MPP+ in female, but not male rats. AB - The effects of in vivo estrogen treatment upon MPP(+)-induced dopamine (DA) release were determined using in vivo microdialysis in female and male rats. Ovariectomized female rats were implanted or not with an estrogen pellet (0.1 mg, 17beta estradiol) and subjected to microdialysis 6 days later. After baseline DA release was determined, 5 mM MPP(+) was infused through the microdialysis probe for one 20-min interval. Perfusion resumed with normal medium for the duration of the experiment. A significant attenuation of MPP(+)-induced DA release was obtained in estrogen-treated females. One week later, striatal DA and dihydroxyphenylacetic acid (DOPAC) concentrations were determined for the lesioned and non-lesioned striata of each animal. MPP(+) infusion significantly decreased striatal DA concentrations, however, there was no effect of estrogen treatment on striatal DA depletion. This experiment was repeated using orchidectomized male rats treated with 0, 0.1, or 5 mg estradiol. In contrast to the females, no differences in MPP(+)-induced DA release were seen among these males, and there was no significant effect of the varying estrogen treatments on striatal DA or DOPAC concentrations. These results demonstrate that in vivo estrogen treatment attenuates MPP(+)-induced striatal DA release in gonadectomized female, but not male, rats. PMID- 10841893 TI - Transporter reversal as a mechanism of glutamate release from the ischemic rat cerebral cortex: studies with DL-threo-beta-benzyloxyaspartate. AB - Elevated levels of the excitotoxic amino acids, glutamate and aspartate, have been implicated in the pathogenesis of neuronal injury and death induced by cerebral ischemia. This study evaluated the contribution of reversed high affinity, Na(+)-dependent, glutamate transport to the ischemia-evoked release of glutamate and aspartate using DL-threo-beta-benzyloxyaspartate (DL-TBOA), a newly developed competitive, non-transported blocker of the EAAT 1-3 transporters. Changes in the extracellular levels of these and other amino acids, and of glucose and lactate in cerebral cortical superfusates during four-vessel occlusion-elicited global cerebral ischemia were examined using a cortical window technique. Basal and ischemia-evoked amino acid, glucose and lactate efflux were compared in control versus DL-TBOA (100 microM; applied topically for 35 min prior to ischemia) animals. Twenty minutes of ischemia caused large increases in aspartate, glutamate, GABA and taurine effluxes into cortical superfusates, with non-significant effects on the efflux of glycine, glutamine, alanine and serine. Application of DL-TBOA caused a 2-fold increase in basal, preischemic, extracellular glutamate levels, but did not affect those of the other compounds. In the presence of DL-TBOA, ischemia-evoked release of aspartate, glutamate, taurine and glutamine was significantly reduced; that of the other amino acids was not affected. The ischemia-evoked declines in glucose were significantly attenuated, and lactate release was enhanced above that in control animals. The amino acid data are interpreted as indicating that aspartate and glutamate releases were reduced as a consequence of DL-TBOA inhibition of reversed transport by high-affinity, Na-dependent carriers, predominantly involving the glial EAAT 2 transporter. The reduction in ischemia-evoked taurine release is interpreted as being due to a decrease in cell swelling prior to and during the initial phase of ischemia due to reduced entry of the Na(+), and other ions, associated with a decreased glutamate uptake. Glucose-sparing and availability for lactate formation would also result from a reduced glutamate/Na(+) uptake. These results indicate that reversed transport, primarily from glial cells by the EAAT 2 carrier, is responsible for a substantial (42 and 56%) portion of the ischemia-evoked increase in extracellular glutamate and aspartate levels, respectively. As a potent, competitive, non-transported blocker of high-affinity, Na(+)-dependent, glutamate transporters, DL-TBOA promises to be a valuable new compound for the study of glutamatergic mechanisms. PMID- 10841895 TI - Zinc-enriched boutons in rat spinal cord. AB - The rat spinal cord reveals a complex pattern of zinc-enriched (ZEN) boutons. As a result of in vivo exposure to selenide ions, nanosized clusters of zinc selenide are created in places where zinc ions are present, including the zinc containing synaptic vesicles of ZEN boutons. The clusters can be silver enhanced by autometallographic (AMG) development. A description of the ZEN bouton patterns is presented and discussed. The distribution of ZEN boutons could indicate that these terminal systems have a differentiated influence on sensory and motor systems. PMID- 10841894 TI - Trophic interactions between brain-derived neurotrophic factor and s100beta on cultured serotonergic neurons. AB - Brain-derived neurotrophic factor (BDNF) and S100beta stimulate serotonergic neurons in fetal rat raphe primary cultures grown under serum-free conditions. BDNF (50 ng/ml) treatment for 3 h enhanced S100beta immunoreactivity in both raphe and hippocampal glial cells. Combined treatment with BDNF and S100beta for 3 days increased the soma area of 5-HT neurons, but not the neurite length. Our results suggest that BDNF and S100beta, which regulate different signal transduction cascades, interact to exert complimentary effects on neuronal maturation by acting sequentially, not concurrently. PMID- 10841896 TI - Antinociceptive effect of cilnidipine, a novel N-type calcium channel antagonist. AB - We investigated the antinociceptic effects of cilnidipine, a dihydropyridine derivative which acts on both L- and N-type voltage-dependent calcium channels, in mice. Intrathecally injected cilnidipine showed significant analgesic effect in formalin test. Cilnidipine significantly suppressed N-type currents in dorsal root ganglion (DRG) cells. Our findings apparently support the idea that cilnidipine attenuates synaptic neurotransmission by inhibiting N-type calcium channels in DRG neurons. PMID- 10841897 TI - Quantitative immunogold evidence that glutamate is a neurotransmitter in afferent synaptic terminals within the isthmo-optic nucleus of the pigeon centrifugal visual system. AB - A quantitative electron microscopic analysis of glutamate (GLU) immunoreactivity using the post-embedding immunogold technique was carried out within the isthmo optic nucleus (ION) of the pigeon centrifugal visual system (CVS). Measurements were performed in each of eight different categories of axon terminals, including those that were GABA-immunoreactive (-ir), considered representing control profiles and identified using a single or double-label immunocytochemical procedure. The results demonstrated that the glutamate immunogold particle densities for both mitochondrial and vesicular pools and for total surface area of bouton profiles were significantly higher in P1a, P1b and P2b terminals and not significantly different in P4 and P5 terminals compared to those recorded in control GABA-ir terminals (P2a, P2c, P3). Moreover, the values measured in GLU-ir positive profiles were all significantly higher than in either P4 or P5 terminals. The results suggest that tectal neurons, which provide the main input to the ION cells, are either inhibitory GABA-ir possibly associated with P2c and/or P3 terminals or excitatory GLU-ir via P1a, P1b and P2b terminals. Such differential effects of tectal afferents may be the basis for the modulation of centrifugal activity and consequently of end target retinal ganglion cell responses. The data are relevant to hypotheses implicating the avian CVS in mechanisms of selective enhancement of visual attention to either novel or meaningful stimuli within the visual field. PMID- 10841898 TI - Fentanyl, a upsilon-opioid receptor agonist, phase shifts the hamster circadian pacemaker. AB - The phase-shifting effects of the mu-opioid receptor agonist fentanyl on the circadian timing system were investigated in the hamster. Fentanyl injections during the mid-subjective day induced phase advances of the hamsters' wheel running activity rhythm. The shifts were not accompanied by an increase in locomotor activity but instead a decrease of activity was often observed. A dose response curve indicated that with increasing dosage, the response probability increased, while the magnitude of the induced shift remained stable. The present data suggest that there is some role for opioid regulation of the circadian system. PMID- 10841899 TI - Fos induction in the brain of mice exhibiting behavioral abnormalities following administration of allylnitrile or crotononitrile. AB - Allylnitrile and crotononitrile induce behavioral abnormalities in mice. To explore the possible involvement of the vestibular system in these behavioral abnormalities, the expression of Fos protein, used as an indicator of neuronal activity, was examined within various brain structures in allylnitrile-, crotononitrile- and vehicle-treated mice. In each nitrile-treated mouse, Fos expression was observed in brain structures, which were divided into two groups. The structures in group 1 showed Fos expression between 1.5 h and 2 days postdosings, and in those in group 2 expression remained for up to 30 days postdosing. As most of these structures, especially in group 2, were identical to some Fos-positive structures observed after unilabyrinthectomy, the present results indicate that each nitrile induces Fos expression by causing a change in the peripheral vestibular system, resulting in behavioral abnormalities. PMID- 10841900 TI - The importance of free hydroxyl radicals to hypoxia preconditioning. AB - Hypoxia preconditioning states that a sublethal hypoxia period will afford neuroprotection against a second harmful event. In our experiments, we carried out a procedure for the development of hypoxia preconditioning in adult male Wistar rats using hypoxic exposure (9% O(2); 91% N(2)) for 1 h. The protection against pentylenetetrazol (PTZ)-induced seizures was studied. For this, rats were tested by a single injection of PTZ (55 mg/kg i.p.) on days 1-21 after hypoxia exposure. The hypoxia exposure significantly prevented the development of acute PTZ convulsion at different times after hypoxia. The present study was designed to determine the effect of N-t-butyl-alpha-phenylnitrone (PBN), an electron trapping agent and free radical scavenger, on hypoxia preconditioning against PTZ seizures 7 days after hypoxia exposure. PBN abolished the protective action of hypoxia exposure. The generation of free hydroxyl radicals in the brains of animals exposed to hypoxia was determined in a second experiment. For this purpose, the rats were i. p. pretreated with 30 mg/kg PBN and NaCl, respectively, 20 min before the start of hypoxia exposure. Forty-five minutes later the rats were i.p. injected with 300 mg/kg sodium salicylate and once again exposed to hypoxia for 15 min. Immediately after that the animals were decapitated and the free hydroxyl radicals and the salicylate content were estimated in the whole brain without cerebellum. Hypoxia preconditioned animals pretreated with NaCl showed a significantly higher extent of free hydroxyl radicals in the brain compared with PBN-injected preconditioned animals and with naive and sham exposed controls. The results pointed out that the generation of free reactive oxygen species under hypoxic conditions in the brain is involved in the development of the hypoxic preconditioning phenomenon. PMID- 10841901 TI - Effect of hyperventilation on brain tissue oxygenation and cerebrovenous PO2 in rats. AB - Previous studies have shown that cortical tissue oxygenation is impaired during hyperventilation. However, it is important to quantify the effect of hyperventilation on brain tissue PO(2) and cerebrovenous PO(2) simultaneously especially since cerebral venous oxygenation is often used to assess brain tissue oxygenation. The present study was designed to measure the sagittal sinus PO(2) (PvO(2)), brain tissue PO(2) in the thalamus (PtO(2)), and brain temperature (Bt) simultaneously during acute hyperventilation. Isoflurane-anesthetized rats were hyperventilated for 10 min during which time the arterial carbon dioxide tension (PaCO(2)) dropped from 40.3+4.9 mmHg to 23.5+2.8 mmHg. PtO(2) declined from 26.0+/-4.2 mmHg to 14.8+/-5.2 mmHg (P=0.004) while brain temperature decreased from 36.5+0.3 degrees C to 36.2+0.3 degrees C (P=0.02). However, PvO(2) and arterial blood pressure (BP) did not change during hyperventilation. The maintenance of PvO(2) when perfusion is thought to decline and PtO(2) decreases suggests that there may be a diffusion limitation, possibly due to selective perfusion. Therefore, cerebrovenous PO(2) may not give a good assessment of brain tissue oxygenation especially in conditions of acute hyperventilation, and deeper brain regions other than the cortex also show impaired tissue oxygenation following hyperventilation. PMID- 10841902 TI - Epitopes of human brain acetylcholinesterase. AB - The main purpose of the present work was to identify B-cell epitopes on human brain acetylcholinesterase (AChE) by the synthetic peptide approach. Five hundred and seventy-four decapeptides comprising amino acids No. n to n+9 (where n denotes the residue number of the 583 amino acids in the primary structure of human brain AChE and is an integer in the range 1-574) were synthesized, using the multipin combinatorial chemical synthesis technique, and biotinylated. Epitopes of human brain AChE were detected by enzyme-linked immunosorbent assay (ELISA) and compared with the predicted epitopes of human AChE by 'Goldkey' software. Among 574 synthetic decapeptides, 47 decapeptides at 11 antigenic regions showed immunoreactivity with mouse anti-human brain AChE polyclonal antibodies. The minimum sequence of epitope was defined at every antigenic region explored. The locations and sequences of the former ten continuous epitopes at the 11 antigenic regions of the human brain AChE had been identified as follows: TPVLVWIY (112-119), RTVLVSMNY (143-151), LLDQRLALQW (173-182), RRATQLAH (246 253), VFRFSFVPV (294 approximately 302), KDEGSYFLVY (332-341), RVYA (424-427), LMRY (476-479), KAPQWPPY (496-503), GLRAQACAFW (523-532). The rate of hits of the predicted epitopes from the software came out at 33%. In our work, the epitopes of human AChE have been mapped by purified polyclonal antibody at eleven distinct sites in the primary structure. PMID- 10841903 TI - From uroscopy to molecular analysis--improving diagnostic information from urine analysis. PMID- 10841904 TI - From uroscopy to urinalysis. AB - Within several decades, 1830-1860, a theoretical system which had left its mark on the practice and rationale of medicine for more than 2000 years, the theory of the four humors, collapsed, taking with it uroscopy as the most important diagnostic tool of the physician's trade. What was the prior and subsequent nature of 'science'? At the transition from uroscopy to urinalysis, contemporary writings manifest a change of paradigm which is unparalleled in the history of medicine, exemplified by the black urine of melanoma. The upheaval changed the doctor-patient relationship ('from bedside medicine to laboratory medicine') and brought forth a 'new type of scientist'. After 160 years of further development, it is time to abandon the triumphant stance of scientific medicine over traditional uroscopy and to restore the role of the latter in terms of what it meant for many centuries in the paradigm of human action: an area of projection of sensory physical experience of the world as well as a platform of medical competence and therapeutic interaction, embedded in the awareness of the limits of human existence. PMID- 10841905 TI - Diagnosis of diabetes mellitus at the hospital of Venice in 1863. AB - In 1674 Thomas Willis reported that the presence of urine 'as sweet as honey' was the pathognomonic sign of diabetes mellitus. In the 19th Century several reactions for the detection of glucose in urine were proposed and glucose measurement became common in the laboratories that were being set up in Europe. A case of diabetes mellitus, diagnosed by Namias, the head of the Women's Section of the Medicine Department of Venice Hospital, was reported in 1863 in the 'Giornale Veneto di Scienze Mediche' which contains clinical and laboratory information. A 34-year-old woman was admitted to the hospital for polydypsia, polyuria, bulimia and fatigue. Urine was weighed for 2 months (2-10 kgday(-1)) and the relative density ranged from 1.045 to 1.038. Glucose was measured in the urine using Moore, Trommer and Fehling reagents. A few days after admission a urine sample showed 7.69 parts/100 parts of urine and a blood sample showed 547 mg of glucose/100 g of serum. The assays were carried out in the Clinical Laboratory of Venice Hospital, founded in 1863, directed by Giovanni Bizio, one of the first chemists who graduated at Padua University. In 1863 chemical analyses were commonly carried out in Venice as in the other parts of Habsburg empire. PMID- 10841906 TI - Characteristics of progressive renal disease. AB - Virtually all renal diseases progress to terminal renal failure relatively independently of the initial disease. Arresting the rate of the deterioration of kidney failure has a great impact on reducing the number of patients reaching the stage of expensive renal replacement therapy. Understanding the mechanisms of the progression of kidney disease has greatly been improved during recent years. The nature of the progressive renal damage with various etiologies includes various well-known factors where hemodynamics, renin-angiotensin system (RAS) and progressive proteinuria play the central roles. Proteinuria has to be shown as an independent risk factor for renal disease progression. Also, disturbances in lipid metabolism as well as the later structural lesions contribute to the progression. Various modalities have been used for the prevention of progressive renal disease, e.g. low-protein diet, antihypertensive therapy, antifibrotic therapy. Many recent experimental and clinical studies have shown that besides the systemic blood pressure lowering effect, RAS blocking agents provide renal protective effects via direct, hemodynamic, and indirect, non-hemodynamic, pathways: (1) lowering intraglomerular capillary hydraulic pressure, and increasing the glomerular ultrafiltration coefficient; (2) lowering proteinuria; (3) lowering hyperlipidemia; (4) diminishing kidney growth; (5) diminishing infiltration of macrophages; (6) downregulation of proinflammatory cytokines. Therefore, RAS blocking agents are widely prescribed not only for antihypertensive but also for renoprotective purposes in diabetic and non diabetic nephropathies. PMID- 10841907 TI - Progressive renal disease: does the quality of the proteinuria matter or only the quantity? AB - Proteinuria is now accepted to be not just a sign of renal disease but also a contributory factor to the development of progressive tubulointerstitial fibrosis. Excellent correlations between the degree of proteinuria and rate of decline of glomerular filtration rate have been demonstrated. What has been investigated less is whether the type of protein found in the urine is important. Using transformed and primary human proximal tubular epithelial cells, we have investigated the binding of albumin and retinol binding protein to plasma membrane preparations and studied the response of the intact cells to increasing concentrations of these same proteins. We have preliminary evidence for differences in the pattern of binding of these two proteins to the plasma membrane receptors and also for differential release of pro-inflammatory cytokines from intact cells. These in vitro results, along with those of other groups, and some recent clinical findings suggest that the quality of proteinuria may play a role in the early development of interstitial fibrosis. Furthermore, the use of such in vitro model systems based on human proximal epithelial cell culture can provide a means of evaluating the potential significance of different markers of tubular damage. PMID- 10841908 TI - Developments in the assessment of glomerular filtration rate. AB - The assessment of the glomerular filtration rate (GFR) is the most commonly used test of renal function. The accepted reference procedure employs an exogenous clearance marker whilst the most popular test is that of serum or plasma creatinine. All of these tests have limitations, although the surrogate endogenous markers are the most practical. Cystatin C, a low molecular weight protein which can be measured by light scattering immunoassay, possesses many of the attributes required of the ideal GFR marker. Data on reference ranges indicate that circulating cystatin C levels reflect the variation in GFR throughout life and the marker demonstrates a better correlation with the reference procedure than serum creatinine. PMID- 10841909 TI - Clinical value of cystatin C determination. AB - It has been reported that cystatin C (cys-C) is elevated in patients with malignant disease. In order to investigate whether this phenomenon is linked to or independent of renal function, and at the same time examine the role of this marker in other pathological situations, cys-C concentrations were compared with 24-h creatinine clearance values in three groups of patients; the first group were undergoing treatment for malignant disease, the second group were renal transplant patients and the third randomly taken from patients for whom a routine creatinine clearance had been requested. Several patients with malignant disease had high cys-C levels without any correspondence to creatinine clearance values. Additionally, although cys-C shows a high sensitivity for detecting impaired glomerular function in renal transplant patients, the specificity was very low, with little discrimination being observed between patients with normal and pathological creatinine clearance levels. In other patients both the sensitivity and specificity of cys-C could be shown to be very good. Thus although cys-C can generally be recommended as a marker of the glomerular filtration rate, there are some patients for whom the clinical relevance is unclear. PMID- 10841910 TI - Diagnostic and prognostic significance of proteinuria selectivity index in glomerular diseases. AB - The proteinuria selectivity index (SI) describes changes of the glomerular permeability for macromolecules. In the present study, we examine the implications of SI as a diagnostic (199 patients) and a prognostic (49 patients) marker in glomerular diseases. Using SI based on alpha(2)-macroglobulin (alpha(2) M-SI) or on IgM (IgM-SI) we found that minimal change nephropathy could be discriminated by low SI values and crescentic necrotizing glomerulonephritis by high SI values compared to other diseases. SI based on IgG (IgG-SI) was less useful in determining specific diagnoses. During a follow-up of 46 months creatinine clearance (Cr cl) decreased 36% in a group of patients with high IgG SI (>0.2) and 38% in a group of patients with high IgM-SI (>1.5(-3)) compared to only 8% in patients with low IgG-SI (10% of their tissues. Thus, the majority of polyps grew isometrically and determinately, virtually ceasing growth by about 30-40 years of age. Coral ages as revealed by skeletal growth rings were similar to those estimated from a growth curve based on field data. The frequency of individuals in each age class on the reef slope decreased exponentially with coral age, indicating high mortality rates when corals were young. The maximum coral age observed in the field population (31 years) was similar to that estimated by application of a population dynamic model (30 years). Calculated rates of growth, mortality and life expectancy for F. granulosa were within the range of those known for other stony corals. Our results reveal a young, dynamic population of this species on Eilat reefs, with high turnover rates and short lifespans. Such information is important for understanding recovery of coral reefs from disturbances, and for application to the management of commercially exploited coral populations. PMID- 10841936 TI - The synthesis of mycosporine-like amino acids (MAAs) by cultured, symbiotic dinoflagellates. AB - We tested the hypothesis that there is a relation between phylotypes (phylogenetic types, as determined by restriction fragment length polymorphism (RFLP) and partial sequence analysis of the small subunit ribosomal RNA gene (SSUrDNA)) and the synthesis of mycosporine-like amino acids (MAAs) by symbiotic dinoflagellates under the influence of ultraviolet radiation (UV-B/A) and photosynthetically active radiation (PAR). We exposed 27 isolates of symbiotic dinoflagellates simultaneously to UV-B/A and PAR, and subsequently determined the MAAs present in cell extracts and in the media. The algae used included 24 isolates of Symbiodinium spp. originating from jellyfishes, sea anemones, zoanthids, scleractinians, octocorals, and bivalves, and three others in the genera Gymnodinium, Gloeodinium and Amphidinium from a jellyfish, an hydrocoral and a flatworm, respectively. In this study, all of the phylotype A Symbiodinium spp. synthesized up to three identified MAAs. None of the 11 cultured phylotypes B and C Symbiodinium spp. synthesized MAAs. The three non-Symbiodinium symbionts also synthesized up to three MAAs. The results support a conclusion that phylotype A Symbiodinium spp. have a high predilection for the synthesis of MAAs, while phylotypes B and C do not. Synthesis of MAAs by symbiotic dinoflagellates in culture does not appear to relate directly to depths or to the UV exposure regimes from which the consortia were collected. PMID- 10841937 TI - Postlarval chromatophores as an adaptation to ultraviolet radiation. AB - It is now well established that ultraviolet radiation (UVR) may have detrimental, even lethal effects on zooplankters. Unlike copepods and other holoplankters, which may avoid UVR by undergoing diel vertical migration, larvae of many decapod crustaceans and fishes recruit to adult populations by remaining in near-surface waters during the daytime. Consequently, they are exposed to biologically damaging UVR. A possible adaptation in these larvae is chromatophores, which may absorb UVR by expanding in high light environments. The supposition is that expanded chromatophores more effectively absorb UVR, but there is some fitness cost to having expanded chromatophores in low light environments. Since the ratio of visible light to UVR in the water column changes as result of season, latitude, dissolved organic carbon, and a host of other factors, the benefits of chromatophores would be maximized if they responded specifically to UVR. The purpose of this study was to determine whether the chromatophores of crab postlarvae (megalopae) could expand in response to UVR. Megalopae of two species of crabs (Cancer oregonensis, Telmessus cheiragonus) were collected from large surface-swarms during mid-day as they recruited onshore in early May 1998 at Friday Harbor, Washington, USA. Dark-adapted megalopae (held in the dark for 8 h before experiments) were exposed to UVR (UVBR+UVAR, 280-400 nm), UVAR (320-400 nm), and light (400-1700 nm) in the laboratory. Chromatophores expanded after only minutes of exposure to UVR, UVAR, and light for both species. Two alternative hypotheses may explain why both harmful and comparatively benign wavelengths stimulated chromatophores to rapidly expand. First, larvae may not distinguish among different wavelengths, which, if true, would increase the vulnerability of these larvae to intensifying UVBR due to ozone depletion. Second, chromatophores have functions other than blocking UVR, such as crypsis and thermoregulation, and must respond to light for these other functions to operate. PMID- 10841938 TI - The intertidal distribution of two algal symbionts hosted by Anthopleura xanthogrammica (Brandt 1835). AB - This paper quantifies the spatial distribution of zooxanthellae (ZX) and zoochlorellae (ZC), two algal symbionts common to the temperate anemone, Anthopleura xanthogrammica, in relation to shore height. Anemones in tidepools and crevices had varying algal proportions: >0.90 ZC (green anemones), 0.10 to 0.90 ZX (mixed), and >0.90 ZX (brown). Brown anemones are primarily found in the high intertidal and the upper region of tidepools. Mixed anemones are most common at intermediate shore heights and green anemones are exclusive to the low shore and at increasing depth in tidepools. Microhabitat was also important to algal proportion, as anemones in crevices had greater proportions of ZC than anemones in tidepools at the same shore height. In a reciprocal transplant experiment, A. xanthogrammica were moved between high and low shallow tidepools. All anemones moved from a low to a high tidepool exhibited a shift from ZC to ZX populations, while the anemones transplanted from high to low tidepools maintained ZX dominance. This is the first documentation that field algal populations can shift from ZC to ZX in Anthopleura. The field survey and transplant study results support the hypothesis that the relative abundance of ZX and ZC in A. xanthogrammica is influenced by the environmental gradient associated with shore height and microhabitat. PMID- 10841939 TI - Effect of food and sediment pre-treatment in experiments with a deposit-feeding amphipod, Monoporeia affinis. AB - We experimentally investigated the effects of different pre-treatments of the sediment, and the effect of daily addition of fresh phytoplankton, on the growth and survival of 1-year-old (1+) individuals of the deposit feeder Monoporeia affinis (Amphipoda). We used three different types of sieved sediment: pre-frozen muddy clay, non-pre-frozen muddy clay, and fine sand. The muddy clay contained phytoplankton originating from the surface sediment sampled in the field during the late spring bloom. No phytoplankton was initially present in sand. The experiment lasted for 18 days. M. affinis responded to the daily phytoplankton addition by increasing growth. Phytoplankton addition had no significant effects on the survival of M. affinis. Upon phytoplankton addition, the sandy and non frozen muddy clay gave similar growth and survival responses. In contrast, the pre-frozen sediment resulted in significantly lower growth and survival. The growth was negative in all treatments without phytoplankton. Thus, the high initial chlorophyll content in the muddy clay was not of sufficient quality or concentration to allow a positive growth response in M. affinis. The growth of M. affinis was significantly correlated with the reduction of the chlorophyll. Our results indicated that M. affinis is capable of assimilating settled phytoplankton with no, or only a few days' time delay. PMID- 10841940 TI - Changes in the acute phase complement component and IL-6 levels in patients with chronic hepatitis C receiving interferon alpha-2b. AB - In order to study the effect of interferon alpha on the levels of acute phase complement proteins in vivo, serum concentrations of C9 and C1-inhibitor (C1-INH) were measured in patients with chronic hepatitis C before and 3 months after the beginning of interferon alpha2b therapy. Serum levels of the activation product of terminal complement pathway, C5b-9, HCV RNA and IL-6 were also determined. IFN alpha treatment significantly (P<0.0001) increased the serum concentrations of both complement proteins. C5b-9 levels were found to significantly decrease during the same period of time. When the patients were divided into responders or non-responders (more or less than 50% decrease in plasma HCV RNA concentrations) C9 and C1-INH levels were elevated only in the responder patients. There was no correlation between the changes of IL-6 levels or the amounts of IFN alpha administrated on one hand, and the changes in the complement protein levels on the other. These findings suggest that the marked increase in the serum concentrations of the acute phase complement proteins is a secondary phenomenon due to the IFN alpha-caused diminution of the viral load and the resulting immune complex-induced complement activation. PMID- 10841941 TI - Strong alpha beta and gamma delta TCR response in a patient with disseminated Mycobacterium avium infection and lack of NK cells and monocytopenia. AB - Infection with atypical mycobacteria occurs mainly in patients with a compromised cellular immune system, in particular in those with a defective T cell or monocyte function. Here we analyzed the specific immune response of an adolescent HIV-negative patient with disseminated mycobacterium avium infection and fatal varizella zoster virus infection. The patient presented with dysplastic hematopoesis of all cell lineage's and a bicytopenia of erythrocytes and leukocytes, but a hematological malignancy could not be found. We found a peripheral lymphopenia and monocytopenia, as well as a lack of NK-cells and B cells. Lymphocytes consisted of 95% T cells, which contained up to 40% of TCR gammadelta+CD4-CD8-T-cells (mainly TCR gamma9delta2), few monocytes and B-cells. Approximately 50% of CD3+ T-cells showed a CD57+ NK-like phenotype. Functional analysis of PBMC revealed a good antigen-specific T cell function if antigen presenting cells were supplemented from a HLA-matched donor. Moreover, a strong M. avium specific cytotoxicity mediated by TCR alphabeta+T-cells could be found in vitro and even ex vivo. In contrast, NK-killing was absent. No evidence for a defect in IL-12 or IFN-gamma production and signaling were found. The data indicate that a strong alphabeta and gammadelta T cell immunity tries to compensate for a deficient monocyte and NK cell function in this patient. PMID- 10841942 TI - Comparison of signaling pathways involved in apoptosis of a thymocyte hybridoma triggered by a rat thymic medullary epithelial cell line, dexamethasone or T-cell receptor cross-linking. AB - Using an in vitro co-culture assay we found that a rat medullary thymic epithelial cell (TEC) line (TE-R2.5) induces apoptosis of the BWRT8 thymocyte hybridoma (TH) (CD4(hi)CD8(low) alphabetaTCR(hi)). TH apoptosis induced by this TEC line was predominantly mediated by direct cell-cell contacts and was potentiated by cross-linking of the T cell receptor (TCR) by R73 monoclonal antibody (mAb). Dexamethasone (Dx) also triggered TH apoptosis but inhibited death of these cells induced by TE-R2.5 cells or immobilized R73 mAb. The TEC induced apoptosis was independent of the LFA-1/ICAM-1 interaction but partly depended on a novel 29 kDa molecule expressed on TE-R2.5 cells. All three types of TH apoptosis were followed by the cleavage of poly-(ADP-ribose)-polymerase and were blocked by a caspase inhibitor Z-Val-Ala-Asp(OMe)-CH(2)F.PKC stimulation by phorbol myristate acetate interfered with the TH apoptosis induced by TE-R2.5 and Dx, but did not modulate the effect of R73 mAb. On the contrary, inhibition of calcineurin with cyclosporine A did not influence the apoptosis induced by TE R2.5 and Dx, but completely prevented the R73-triggered TH cell death. The TE R2.5-mediated BWRT8 apoptosis was suppressed by Na-orthovanadate, an inhibitor of protein tyrosine phosphatases (PTP) as well as by genistein, a protein tyrosine kinase (PTK) inhibitor, while both compounds potentiated the effect of Dx. Blocking PTP, but not PTK decreased the proapoptotic effect of R73 mAb. These results, including those using a BWRT8 subclone (BWRT8-MDP.2) which is resistant to TCR-triggered apoptosis, but sensitive to apoptosis stimulated by TE-R2.5 and Dx, indicate that TE-R2.5-induced TH apoptosis in our model is different from apoptosis in other TEC co-culture models, published so far. PMID- 10841943 TI - Influence of synthetic peptide corresponding to the ACTH-like sequence of human immunoglobulin G1 on activity of murine thymocytes and peritoneal macrophages. AB - The purpose of the present study was to investigate properties and mechanism of action of the synthetic adrenocorticotropin (ACTH)-like peptide VKKPGSSVKV, corresponding to the sequence 11-20 of the variable part of human immunoglobulin G1 (IgG1) heavy chain. The ACTH-like peptide was shown to act as an immunosuppressive agent in vitro: it inhibits the blast transformation of mouse thymocytes and reduces the spontaneous motility of mouse peritoneal macrophages as well as their bactericidal activity against Salmonella typhimurium 415 virulent strain bacteria. High affinity receptors for the ACTH-like peptide were found on thymocytes and macrophages and shown to be at the same time the receptors for ACTH. The kinetic characteristics of the ACTH-like peptide and 125I labeled ACTH (13-24) (ACTH 'address segment') specific binding to the receptors were determined. It was found that the ACTH-like peptide binding to the receptors on target cells is accompanied by an increase in both adenylate cyclase activity and intracellular cAMP content. PMID- 10841944 TI - Functions of myeloid and lymphoid dendritic cells. AB - The bone marrow derived dendritic cell (DC) is an essential antigen presenting cell (APC) for the initiation of primary, T cell based immune responses. DC are a heterogenous haematopoietic lineage, in that many subsets from different tissues show different surface phenotypes, but the ability to stimulate antigen specific naive T cell proliferation appears to be shared between these DC subsets. It has been suggested that the so called myeloid and lymphoid-derived subsets of DC perform distinct stimulatory or tolerogenic functions. However, recent data has blurred this apparent distinction of DC subset function and shown that both subsets are at least capable of stimulatory and possibly even tolerogenic functions. Thus, the immunoregulatory potential of DC may depend less on ontology than on recent activatory or downregulatory stimuli. PMID- 10841945 TI - Role of follicular dendritic cells in the apoptosis of germinal center B cells. AB - Follicular dendritic cells (FDCs) provide the most obvious source of antigens, which are essential for the differentiation of GC B cells. It has been reported that most proliferating B cells in germinal centers undergo apoptosis. Quantitative histology shows macrophages with apoptotic debris throughout the germinal center, the highest frequency of these cells being found in the dense FDC network. Based on these findings, we hypothesized that FDC may be involved in an apoptotic pathway of the germinal center B cells. To prove this hypothesis, we performed double immunohistochemical analysis using anti-FDC mAb and peanut agglutinin (PNA), with their respective TUNEL kits. Collated data showed that a great proportion of the apoptotic cells, most of which were positive for PNA, were in close contact with FDC, which indicated an interaction between FDC and B cells in the apoptotic pathway. Further studies using double immunohistochemical staining and FACS analyses demonstrated the expression of Fas-ligand (FasL) in a subset of the FDC. These results suggest that FDC may play a role in the apoptosis of germinal center B cells via Fas-FasL interaction. PMID- 10841946 TI - Trafficking of major histocompatibility complex class II molecules in human B lymphoblasts deficient in the AP-3 adaptor complex. AB - The major histocompatibility complex class II subunits (MHC-II) alpha and beta assemble with the invariant chain (Ii) in the endoplasmic reticulum and are transported to endosomal-lysosomal organelles known as MHC class II compartments (MIICs). Although it has been shown that two dileucine-based signals in the cytosolic tail of Ii, as well as a dileucine-based signal in the tail of the beta chain mediate sorting to MIICs, the molecular mechanisms by which alphabetaIi complexes are sorted have yet to be resolved fully. The AP-3 adaptor complex stands out as a particularly good candidate for mediating this targeting because: (i) it has a proven role in the trafficking of membrane proteins to lysosome related organelles; and (ii) it has the ability to interact with dileucine-based signals in vitro. To investigate the potential role of AP-3 in transport of MHC II to MIICs, we have examined MHC-II trafficking in human B-lymphoblast lines from patients with Hermansky-Pudlak syndrome type 2 (HPS-2), which are deficient in the AP-3 complex. Pulse-chase analyses revealed no significant alteration in the kinetics of synthesis and degradation of either MHC-II subunits or Ii. Moreover, we observed neither impairment of the formation of compact SDS resistant alphabeta dimers, nor delay in the appearance of a conformational epitope indicative of a mature, Ii-free alphabeta dimer. Finally, we demonstrated that in HPS-2 patients' cells, there was no delay in the expression of the alphabeta dimers on the cell surface. Thus, AP-3 does not seem to be essential for normal trafficking of MHC-II. These findings have important implications for HPS-2 patients, because they suggest that the recurrent bacterial infections suffered by these patients are not likely due to impaired antigen processing and presentation by MHC-II. PMID- 10841947 TI - Antigen dose defines T helper 1 and T helper 2 responses in the lungs of C57BL/6 and BALB/c mice independently of splenic responses. AB - To investigate the effect of antigen dose on immune response, C57BL/6 and BALB/c mice were sensitized with aluminum hydroxide gel (alum)-precipitated ovalbumin (OVA) then challenged with aerosolized OVA. Low-dose sensitization (less than 8 microg of OVA) elicited T helper 2 (Th2)-type immunoglobulins (Igs) secretion from C57BL/6 mice, including high levels of serum IgE, IgG1 and low levels of IgG2a, while BALB/c mice secreted T helper 1 (Th1)-type Igs, including low levels of IgE, IgG1 and high levels of IgG2a. In contrast, high-dose sensitization (more than 50 microgram) elicited Th1-type Igs secretion in C57BL/6mice, while BALB/c mice exhibited Th2-type Igs secretion. Furthermore, the number of eosinophils infiltrating into the lungs of low-dose OVA-sensitized C57BL/6 mice was significantly greater than in BALB/c mice sensitized with the same amount of OVA. Only a very high dose of OVA (1 mg) could induce greater eosinophil infiltration into the lungs of BALB/c mice compared with C57BL/6 mice. Additionally, low-dose sensitization generated Th2-type cytokines, including high levels of interleukin (IL) -4, IL-5 and a low level of interferon-gamma (IFN-gamma) in the lungs of C57BL/6 mice, while BALB/c mice generated Th1-type cytokines in their lungs, including low levels of IL-4, IL-5 and a high level of IFN-gamma. In contrast, high-dose sensitization elicited Th1-type cytokines production in the lungs of C57BL/6 mice, while BALB/c mice generated Th2-type cytokines in their lungs. Interestingly, splenocyte cultures from C57BL/6 mice produced Th1-type cytokines, while cultures from BALB/c mice produced Th2-type cytokines regardless of OVA sensitization dose (100 ng-1 mg). These results indicate that C57BL/6 and BALB/c mice have different susceptibilities to OVA-sensitization and OVA-induced pulmonary eosinophilia regulated by Th1- and Th2-type cytokines, independent of splenic Th1- and Th2-type cytokines production. PMID- 10841948 TI - Molecular analysis of CD26-mediated signal transduction in T cells. AB - CD26 or dipeptidylpeptidase IV (DPP IV) is a cell surface protease involved in T cell activation. It is a type II transmembrane glycoprotein consisting of a large extracellular part, a single transmembrane region and a short cytoplasmic tail without any common signalling motifs. To eluciate the mechanisms involved in CD26 mediated signalling we have constructed C-terminal deletion mutants of the human CD26 molecule and transfected them into murine T cell hybridomas. Stimulation experiments show that most of the extracellular part of CD26 can be deleted without affecting its costimulatory activity. The membrane proximal glycosylation rich region of CD26 is sufficient to transduce costimulatory signals. Activation of T cells via CD26, however, is not mediated by the important T cell receptor associated adaptor proteins LAT and TRIM as shown in colocalization assays. PMID- 10841949 TI - Histamine suppresses neopterin production in the human myelomonocytoma cell line THP-1. AB - Histamine, an important inflammatory mediator in allergic diseases and asthma, was reported to have modulatory effects on T cells by down-regulating Th1-type cell cytokines like interleukin 2 (IL-2) and interferon-gamma (IFN-gamma). In this study we examined the effect of histamine and the histamine-receptor antagonists cimetidine and diphenhydramine on the production of neopterin after stimulation with IFN-gamma in the myelomonocytoma cell line THP-1. Increasing concentrations of histamine markedly suppressed IFN-gamma induced neopterin formation. Simultaneous preincubation of THP-1 cells with histamine, IFN-gamma and different concentrations of the H(2)-receptor antagonist cimetidine showed a clear antagonizing effect on neopterin formation. In contrast, the H(1)-receptor antagonist diphenhydramine was not able to abrogate the suppressive effect of histamine on neopterin production. Our results suggest, that histamine may be a potent inhibitor of effects or mechanisms induced by IFN-gamma in monocytes/macrophages. Cimetidine, and possibly other H(2)-receptor antagonists, may reverse down-regulatory actions of endogenously formed histamine on activated monocytic cells. PMID- 10841950 TI - Detection of alternatively spliced variant messages of Fas gene and mutational screening of Fas and Fas ligand coding regions in peripheral blood mononuclear cells derived from silicosis patients. AB - Silicosis is clinically characterized not only by respiratory disorders but by immunological abnormalities such as the appearance of autoantibodies and complications of autoimmune diseases. Dysregulation of apoptosis, particularly in the Fas/Fas ligand (FasL) pathway, has been considered to play a role in the pathogenesis of autoimmune diseases. It has been found that serum soluble Fas (sFas) levels are elevated in silicosis patients (SIL) and the sFas message is dominantly expressed in peripheral blood mononuclear cells (PBMC) derived from these individuals. In the present study, one tried to detect alternatively spliced variant messages including typical sFas message and found four that were highly and frequently expressed, and which possess a signal peptide domain, but not transmembrane and signal transducing domains, in PBMC derived from SIL. Functional mutations were not detected in Fas and FasL genes in silicosis PBMC. Still, alternative spliced variants of the Fas gene including typical sFas message appear to play an important role in the immunological dysregulation in SIL. PMID- 10841951 TI - Nasopharyngeal teratomas and dermoids: a review of the literature and case series. AB - OBJECTIVES: Review the clinical differences between nasopharyngeal (NP) true teratomas and dermoids, with the addition of a case series to the literature. Additionally, review the impact of prenatal diagnosis on the management and outcome of these lesions. STUDY DESIGN: Retrospective review of cases from the Children's Hospital Medical Center (Seattle, WA) and Madigan Army Medical Center (Tacoma, WA) with the histopathologic diagnosis of nasopharyngeal teratoma or dermoid. Review of medical literature for cases reported since 1977, when prenatal diagnosis of these lesions was first reported. Only tumors of the nasopharynx were considered. METHODS: Charts and pertinent literature were reviewed and data presented with respect to age at diagnosis, signs and symptoms, perinatal diagnosis and management, preoperative evaluation, surgical treatment, and outcome. RESULTS: The majority of lesions were diagnosed at birth, with the most common presenting symptom being respiratory distress. Teratomas had a higher incidence of maternal polyhydramnios, preterm birth, need for emergent airway management, and associated congenital abnormalities. Complete surgical excision remains the treatment of choice. Recurrences were rare, occurring only in the teratoma group. Prenatal diagnosis did not have a significant impact on the diagnosis and treatment of these lesions in our review. CONCLUSIONS: Inconsistent use of a standard classification system has made differentiating between NP teratomas and dermoids difficult, although the clinical implications can be critical. NP teratomas have a higher incidence of preterm birth, neonatal airway distress, associated congenital abnormalities, need for more extensive surgical procedures, and recurrence. Prenatal diagnosis has made little impact on the overall diagnosis and treatment of these lesions. PMID- 10841952 TI - Experimental results do not support a gas reserve function for the mastoid. AB - The mastoid is an aerated extension of the middle ear gas pocket whose state of development was shown to be an indicator of past and future otitis media experience. While the function(s) of the mastoid is not known, a number of hypotheses has been advanced to explain the reported association between mastoid size and middle ear disease. These include the hypotheses that, with respect to the middle ear, the mastoid functions as a pressure buffer, a gas reserve, and/or a pressure regulator. In this paper, a physical model of the mastoid is presented that makes specific predictions against which the validity of the hypothesized gas reserve function could be tested. Data from three published clinical experiments were evaluated for consistency with the predictions of the model, and the hypothesis was rejected. Also, when reinterpreted within the context of the model, the published data do not support a pressure-regulating function for the mastoid. PMID- 10841953 TI - A comparison of microbipolar cautery dissection to hot knife and cold knife cautery tonsillectomy. AB - BACKGROUND: Morbidity following tonsillectomy continues to be a major concern to parents, children, and physicians alike. Reduction in post-operative complications, optimal control of pain, and satisfactory return to a normal lifestyle are all important to both family and society. This study compares both the complication and recovery rates after microbipolar dissection (MBPD) technique of tonsillectomy to two well established tonsillectomy techniques: hot knife (HK) and cold knife/cautery (CK/C). METHODS: A total of 200 consecutive patients presenting for tonsillectomy by the first author (MP) were randomized to either undergo MBPD or HK tonsillectomy. Concurrently, an additional two hundred patients were randomized to undergo MBPD or CK/C tonsillectomy by the second author (LB). Patients were prospectively followed for complications including bleeding and dehydration and multiple indicators of recovery rate. RESULTS: Postoperative bleeding of any kind was significantly less in the MBPD group than in the CK/C and HK groups (5 vs 12.4 vs 12.5% (P<0.001). The need for post operative intervention for bleeding, i.e. local control or return to the operating room, was also significantly less in the MBPD group. Return to normal activity occurred 2 days earlier in the MBPD group versus either CK/C or HK (P<0.001). Additionally, earlier recovery was reflected in fewer total doses of pain medication in the MBPD group. Operative time was 3-5 min longer for MBPD (24.2 min) than for CK/C or HK (21.1 and 16.5 min, respectively); blood loss was similar (within 15 cc) among all treatment groups. CONCLUSION: MBPD tonsillectomy compared most favorably to conventional techniques (CK/C and to HK tonsillectomy). Important clinical outcome differences included a lower bleed rate, earlier recovery and fewer days lost from work and school. The financial impact is estimated to be quite favorable. MBPD tonsillectomy is now our preferred method in children. PMID- 10841954 TI - Acquisition of hearing aids and assistive listening devices among the pediatric hearing-impaired population. AB - Sufficient access to health care is of concern to the indigent population in the US and to their health care providers. This study was undertaken to elucidate the rate of the follow-up among lower socioeconomic hearing-impaired pediatric patients who had received a recommendation for hearing aids and/or assistive listening devices. Our question was, would the families' financial situation have a negative effect on the acquisition of hearing aids and assistive listening devices? Fifty patients, age 0-18 years, who had been seen in our clinic over 2 years were evaluated via a telephone survey. The survey consisted of seven questions, including whether or not the devices or aids were obtained, what type was purchased, where the device was being used, and the child's apparent performance with the device. Eighty-two percent of our patients were on TennCare, a state mandated Medicaid HMO system. Two-thirds of these TennCare patients are at or below the poverty level and the remaining one-third is either disabled or uninsurable according to the Aid for Dependent Children (AFDC) with indeterminate income. In addition the TennCare organization did not cover hearing amplification equipment for these children. The study showed that the majority of the patients did follow-up as recommended. Furthermore, this equipment is easily obtainable for the pediatric indigent population due to financial resources available in the community outside the mandated Medicaid system. PMID- 10841955 TI - Screening sinus CT scans in pediatric bone marrow transplant patients. AB - OBJECTIVE: To determine how sinus disease noted on pre-bone marrow transplant (BMT) screening sinus computed tomography (CT) scans relates to subsequent development of clinical and/or radiographic sinusitis and correlates with overall prognosis. METHODS: A retrospective review of medical records, CT scans, and BMT database statistics was performed on all pediatric BMT recipients from January 1992 through December 1997. Fifty-four pre-BMT CT scans were performed on 51 children, aged from 2 months to 17 years. Sinus disease was staged according to criteria established by Lund and Kennedy [V.J. Lund, D.W. Kennedy, Ann. Otol. Rhinol. Laryngol. S167 (1995) 17-21.]. RESULTS: The average age of BMT recipients was 6.8 years. Most common oncologic diagnoses included acute myelogenous leukemia (37%), acute lymphoblastic leukemia (17%), and stage IV neuroblastoma (13%). Screening sinus CT scans were routinely performed 1-3 months prior to BMT. On pre-BMT CT scans 48% of the patients had no evidence of sinus disease, 25.9% mild disease, 9.3% moderate disease, and 16.7% severe disease. Two-thirds (66.7%) of patients with severe sinus disease on pre-BMT CT scans experienced clinical sinusitis post-BMT. In contrast, sinus symptoms were much less common (21.4%) in those with mild disease on CT scan. Overall, 39.3% of patients with sinus abnormalities on pre-BMT CT scans had clinical sinusitis during their post-BMT course, compared to 23.1% of those with normal CT scans pre-BMT. In addition, those patients demonstrating sinus disease on their pre-BMT CT scan were more likely to have radiographic sinusitis post-BMT (25.0%) than those with no disease (7.7%). Seventy-eight percent of those with severe sinusitis had died by 2-year follow up, compared to 69.2% of patients with normal CT scans pre-BMT. CONCLUSIONS: Severity of radiographic sinus disease on pre-BMT CT scans was noted to correlate with clinical and radiographic sinusitis later in the post-BMT course, and was associated with a trend toward decreased survival. Pre-BMT CT scans may be useful in determining which children need early and more aggressive intervention for clinical sinusitis post-BMT. PMID- 10841956 TI - The use of intensity modulated radiotherapy for the treatment of extensive and recurrent juvenile angiofibroma. AB - These case series are presented to describe the application and advantages of intensity modulated radiotherapy (IMRT) for the treatment of extensive and/or recurrent juvenile angiofibroma. Two patients were diagnosed with recurrence at 11 and 13 months postoperatively, and one was surgically unresectable. The affected areas included the base of skull, cavernous sinus, pterygopalatine fossa, infratemporal fossa, posterior orbit and nasopharynx. Highly conformal IMRT was delivered with limited radiation doses to the optic nerves, optic chiasm, brainstem, brain, spinal cord, lens, retina, mandible, and parotid. The total dose delivered to the tumor varied from 3400 to 4500 cGy. The tumor shrunk radiographically in all three cases and there was no endoscopic evidence of disease in two cases at 15 months and 40 months. There was no acute toxicity. Late toxicity was limited to one episode of epistaxis and persistent rhinitis in one patient. In conclusion, IMRT provides several advantages over conventional radiotherapy in the treatment of recurrent juvenile angiofibroma. PMID- 10841957 TI - Cholesteatoma in children: techniques and results. AB - A retrospective analysis of the medical records of all cases of cholesteatoma in children treated between 1981 and 1996 was performed. The charts of 59 children with cholesteatoma were reviewed. A total of 62 ears received surgery over 15 years. The median follow-up period was 5 years. There were 132 operations. An intact canal wall (ICW) procedure was performed in 29% of the ears in the first stage, a canal wall down (CWD) procedure in 37%, a transmeatal atticotomy (TA) in 21%, a tympanoplasty (T) in 6.5% and a myringoplasty (M) in 6.5% of the ears. The ICW procedure had a higher rate of residual/recurrent cholesteatoma than did CWD (P=0.8), TA (P=0.4), T (P=0.5), and M (P=0.05). The CWD procedure had a lower rate than TA (P=0.7), but a higher rate than T (P=0.6) and M (P=0.09). Lastly, TA had a higher rate than T (P=0.7) and M (P=0.1). Auditory results were similar for type II and type III reconstructions (P=0.5). An air-bone gap of less than 20 dB was achieved in 51% of the ears, and 80% had a gap of less than 40 dB. We found a clear difference, although not statistically significant, in the personal rate of recurrent cholesteatoma. It was 26% for surgeons who had performed more than 350 otological surgeries and fell to 15% for the most-experienced surgeon (1715 operations). It rose to 34% for the less experienced surgeons (P=0.8). We recommend that surgery for children with cholesteatoma be reserved for experienced surgeons. PMID- 10841958 TI - Intracordal cyst in a neonate. AB - Vocal cord cysts are rare, and typically present with symptoms of dysphonia in older children and adults. There is often a background of voice abuse. We present the first recorded and photographed case of an intracordal cyst presenting in a neonate. The patient presented with symptoms of episodic stridor, respiratory distress, and feeding difficulties, which resolved rapidly after simple incision of the cyst. Theories for the aetiology of intracordal cysts are discussed. The authors believe that this case lends support to the theory that intracordal cysts are congenital in origin. PMID- 10841959 TI - Congenital maxillary sinus cholesteatoma. AB - Cholesteatoma of the paranasal sinuses is a rare condition. The purpose of this paper is to present a child with a congenital maxillary sinus cholesteatoma. An 18-month-old girl presented with a 4-week history of right cheek and intraoral swelling. Examination revealed a smooth swelling of the right hard palate in association with the facial swelling in the maxillary region. An inferior meatal antrostomy revealed pultaceous debris in the right maxillary antrum and biopsy confirmed a maxillary sinus cholesteatoma. The inferior meatal antrostomy was enlarged to allow exteriorisation of the disease. Recurrence of the disease has not presented on follow-up. An exteriorisation procedure as performed, in child of this age, allows normal facial growth. If recurrence develops then further treatment may be instituted in a more mature facial skeleton. PMID- 10841960 TI - Clinical trials, treatment guidelines and real life. PMID- 10841961 TI - Detecting hibernating myocardium: how and why? PMID- 10841962 TI - Comparison between low-dose dobutamine echocardiography and thallium-201 scintigraphy in the detection of myocardial viability in patients with recent myocardial infarction. AB - AIM: To compare the predictive value of thallium-201 single photon emission computed tomography (SPECT) scintigraphy (Sci) and low-dose dobutamine echocardiography (Dob) in predicting late recovery of dysfunctioning myocardium in patients with recent, uncomplicated myocardial infarction (MI). METHODS AND RESULTS: 19 patients (18 male, aged 58+/-8 years) with recent MI and ejection fraction <50% (35.5+/-8.3%) underwent 5-15 microg/kg per min Dob, rest redistribution Sci and coronary angiography, respectively, 14+/-6, 16+/-7 and 17+/-5 days after MI. On an eleven-segment ventricular model devised to compare Dob and Sci segment by segment, each dysfunctioning ventricular segment was considered viable if it showed recovery of mechanical function at the echocardiographic follow-up, performed 6.3+/-1.5 months after revascularization (five PTCA, five GABG) or medical therapy. Among the 104 dysfunctioning segments, of which 26 (25%) showed recovery at follow-up, Dob and Sci gave a concordant response in 50 (48%, k = 0.13), correctly predicting the recovery (or not) of function in 42. Forty-two of 54 discordant responses were due to segments judged viable only by Sci and which had no recovery at follow-up (of these 37 were akinetic or severely hypokinetic at baseline). At the segment-by-segment analysis, the sensitivity, specificity, and accuracy in predicting recovery of function at follow-up were, respectively, 69, 88 and 84% for Dob as against 88, 36 and 49% for Sci (P<0.001 for both specificity and accuracy, P=NS for sensitivity). CONCLUSION: In patients with recent MI, the specificity of Dob in the detection of myocardium capable of late mechanical recovery is significantly higher with respect to Sci, whereas sensitivity is slightly, not significantly higher for the latter. It is conceivable that Sci detects viable myocardium even if it is transmurally limited to epicardial layers in segments with severely impaired mechanical function in which viability will not affect late recovery of function. PMID- 10841963 TI - Central pulmonary artery anatomy in right ventricular outflow tract obstructions. AB - We reviewed the cine-angiograms of 190 patients with right ventricular outflow tract (RVOT) obstructions for size and anatomy of pulmonary arteries, patent ductus arteriosus (PDA) and major aorto pulmonary collateral arteries (MAPCAs). Patients were grouped into three, Tetralogy of Fallot (TOF) with pulmonary atresia (group 1, N=86), TOF with pulmonary stenosis (group 2, N=97) and 7 cases of pulmonary atresia with intact interventricular septum (group 3). Out of 86 patients in group 1, 49 had PDA alone, 30 had MAPCAs alone, six had both and one had none. In group 2, 31 patients had persistent PDA and one patient had MAPCAS and PDA. A discrete stenosis (DS) of pulmonary artery was seen significantly more in patients with RVOT obstructions associated with PDA compared to patients without PDA (67/84 vs. 5/96). Out of the 84 cases with ducti, 53 had stenosis of the pulmonary artery at the site of ductus insertion. Thus presence of PDA was an important factor in the development of DS. The likely cause of pulmonary artery stenosis in TOF with PDA may be the opposing flows through RVOT and PDA producing a watershed effect at the ductus-pulmonary artery junction. Diffuse hypoplasia of pulmonary arteries (DH) was seen more significantly in RVOT obstructions associated with MAPCAs, compared to other patient groups (19/36 vs. 14/87). These small pulmonary arteries had no discrete stenosis and this diffuse hypoplasia might be the result of inadequate blood flow during intrauterine life [Harikrishnan S, Tharakan J, Titus T, Bhat A, Sivasankaran S, Bimal F, Syam Sunder KR, James, KJ. Central pulmonary artery anatomy in right ventricular outflow tract obstructions. Indian Heart Journal 1997;49:624 (Abstract)[18]]. PMID- 10841964 TI - Protective effect of high density lipoprotein on endothelium-dependent vasodilatation. AB - Low concentrations of high-density lipoprotein cholesterol (HDL-C) have been associated with increased risk of coronary heart disease (CHD) even when the total cholesterol (TC) and triglyceride (TG) levels are not elevated. The mechanism by which HDL confers protection against atherosclerosis remains speculative. Using high-resolution ultrasound, we measured the dilatation changes of brachial arteries during reactive hyperemia and after sublingual glyceryl trinitrate (GTN) in 63 patients with established (CHD) and 45 controls, in which the serum TC level was normal. The results showed that both flow-mediated dilatation (FMD) and GTN-induced dilatation of brachial arteries in patients with CHD were much reduced compared with control group (2.31+/-2.46% vs. 7.43+/-4.10% and 16.41+/-6.15% vs. 22.44+/-8.63%, respectively, P<0.001 for all). Univariate analysis indicated that FMD of brachial arteries was inversely related to age (r= 0.226, P<0.05), hypertension (r=-0.229, P<0.05), baseline diameter (r=-0.299, P<0.01) and LDL-C (r=-0.237, P<0.05) and positively related to HDL-C (r=0.491, P<0.01). GTN induced vasodilatation was inversely related to age (r=-0.216, P<0. 05) and baseline diameter (-0.476, P<0.01). Multiple stepwise regression analyses in two groups taken together showed that HDL-C and age were the independent predictors of the FMD of brachial arteries (beta=0.466, P=0.000 and beta=-0.184, P=0.020, respectively). Baseline diameter was significant predictor of GTN induced vasodilatation (beta=-0.390, P=0.000). The analysis in the group of CHD patients showed that only HDL-C was significantly relate to the FMD of brachial arteries (beta=0.295, P=0.018 ) and in controls that hypertension and HDL-C were significantly relate to the FMD of brachial arteries (beta=-0.395, P=0.004 and beta=0.344, P=0.011, respectively). These finding suggest that endothelium dependent and endothelium-independent vasodilatation are impaired in the patients with CHD. HDL exerts a protective effect on endothelium-dependent vasodilatation in TC being relatively normal population. PMID- 10841965 TI - True shape and area of proximal isovelocity surface area (PISA) when flow convergence is hemispherical in valvular regurgitation. AB - The proximal isovelocity surface area (PISA) method for quantifying valvular regurgitation uses an echocardiographic image with superimposed colour Doppler mapping to visualise the contours of velocity in the blood travelling towards the regurgitant orifice. The flux of blood through the regurgitant orifice is obtained as the product of the area of one of these (presumed hemispherical) contours and the speed of the blood passing through it. However, colour Doppler mapping measures the velocity component towards the echo probe (v cos theta;) rather than speed (v), so that the contours of equal Doppler velocity (isodoppler velocity contours) differ from isospeed contours. We derive the shape of the isodoppler contour surface obtainable by colour Doppler mapping, and show that its area is much less than that of the hemispherical isospeed contour. When regurgitant flux is derived from an appropriate single measure of contour dimension, an appropriate result may be obtained. However, if the true echocardiographic surface area is measured directly, the regurgitant flux will be substantially underestimated. Indeed, the conditions necessary for isodoppler velocity contours to be hemispherical are extraordinary. We should not therefore make deductions from the apparent shape for the convergence zone without considering the principles by which the image is generated. The discrepancy will assume practical significance when increased resolution of colour Doppler technology makes measurement of apparent surface area feasible. Assuming the flow contours are indeed hemispherical, a 'correction' factor of 1.45 would be required. PMID- 10841966 TI - Right ventricular long axis function in adults and children with Ebstein's malformation. AB - OBJECTIVES: To assess right ventricular function in adults and children with Ebstein's anomaly. DESIGN: Prospective study. SETTING: Tertiary referral centre. SUBJECTS: Fifteen patients (8 adults and 7 children) with Ebstein's anomaly and 14 healthy controls. INTERVENTIONS: Transthoracic echocardiography was performed in all. Right ventricular function was assessed from long axis M-mode recordings of the right atrio-ventricular free wall. Total systolic excursion as well as peak shortening and lenghtening rates of the right ventricle were measured. RESULTS: Children and adult patients with Ebstein's anomaly differed in terms of age at diagnosis, the adult group having been diagnosed later 19.8+/-15.8 vs. 5.9+/-6.2 years, P<0.05. Measures of right ventricular long axis function in children with Ebstein's anomaly showed a significantly reduced systolic excursion 1.4+/-0.4 vs. 2.4+/-0.53 cm, P<0.05 and peak lenghtening rate; early 8.04+/-4.3 vs. 11.8+/-2.4 cm/s and late 6.14+/-3.6 vs. 10.6+/-4.3 cm/s, P=0.05 compared to controls. In contrast, measurements of right ventricular long axis function in adults with Ebstein's anomaly showed no significant difference when compared to the control group. Transtricuspid Doppler flow velocities were not different between patient's groups and corresponding controls. CONCLUSION: The right ventricle assessed by this simple, non-invasive technique reveals a significantly reduced systolic and diastolic function in children with Ebstein's malformation compared to controls but a 'normal' right ventricular function comparable to controls in adult patients. Significant right ventricular dysfunction in children with Ebstein's anomaly could account for their earlier presentation. Long term follow up of the right ventricular abnormalities is needed in such children to discover more about the natural history of the disease. PMID- 10841967 TI - Role of the etiology of cardiomyopathies on exercise capacity and oxygen consumption in patients with severe congestive heart failure. AB - Peak oxygen consumption is of great importance for the decision of heart transplantation in congestive heart failure. Moreover, the level of exercise capacity seems to depend on the etiology of congestive heart failure. This study compared 14 heart failure patients with idiopathic dilated cardiomyopathy (group 1) to 14 heart failure patients with cardiomyopathy due to ischemic heart disease (group 2), matched for sex (13 male, one female in each group), age +/-10 years, left ventricular ejection fraction +/-5% and pulmonary artery mean pressure +/-5 mm Hg, to assess exercise capacity and oxygen consumption independently of the age, sex and the level of left ventricular dysfunction. Right ventricular function was also assessed. No difference existed in terms of right ventricular parameters. Maximal exercise parameters were significantly higher in group 1 than in group 2. Peak oxygen consumption was statistically higher in group 1 than in group 2. In the whole population, a significant correlation was found between peak oxygen consumption and right ventricular ejection fraction (r=0. 44, P<0.02) but not between peak oxygen consumption and left ventricular ejection fraction. For similar levels of left ventricular dysfunction, exercise capacity and oxygen consumption appear to be better in idiopathic dilated cardiomyopathy than in ischemic cardiomyopathy, thereby suggesting that functional tolerance of left ventricular dysfunction might depend on the etiology of severe congestive heart failure. PMID- 10841968 TI - The effect of salbutamol on skeletal muscle in chronic heart failure. AB - We performed a randomised placebo-controlled trial to investigate the effects of the anabolic drug salbutamol on skeletal muscle and exercise capacity in chronic heart failure. Twelve patients received salbutamol slow-release 8 mg twice daily or placebo for 3 weeks. We assessed the effect of treatment on exercise capacity, quadriceps muscle bulk, maximal isometric strength and fatigue, respiratory muscle strength, spirometry and 24-h ECG (electrocardiogram). There was no significant change in the muscle indices, exercise time or peak oxygen consumption. The frequency of ventricular arrhythmias and spirometric measurements were also unchanged. Maximal expiratory mouth pressure, measured at total lung capacity and functional residual capacity, increased significantly (+29.7+/-10.6 vs. -0. 5+/-7.5 cm H(2)O [mean+/-S.E.M., change over 3 weeks treatment salbutamol vs. placebo] and +31.2+/-5.4 vs. +0.2+/-4.0 cm H(2)O both P<0.05). Maximal inspiratory pressures showed a trend towards increasing with treatment when measured from either lung volume (-22. 8+/-9.5 vs. -6.2+/-3.6 cm H(2)O, P=0.14 and -21.5+/-7.5 vs. -3.5+/-3. 4 cm H(2)O, P=0.054). Treatment with 3 weeks of salbutamol increases respiratory muscle strength in chronic heart failure but does not improve quadriceps abnormalities or exercise capacity. Salbutamol is unlikely to have a role in treating the muscle abnormalities in chronic heart failure. PMID- 10841969 TI - Abolition of exercise induced positive U-wave after coronary angioplasty: clinical implication. AB - OBJECTIVE: We prospectively studied the course of exercise induced positive U wave before and after percutaneous transluminal coronary angioplasty (angioplasty). BACKGROUND: Negative U-wave in ECG is known to be associated with the myocardial ischemia in the territory of the left anterior descending artery. Positive U-wave needs further evaluation to prove its diagnostic value in localization of coronary artery disease. METHOD: Twenty patients demonstrated exercise induced positive U-wave from a cohort of 730 patients referred because of chest pain. Exercise was carried to > or =90% of target heart rate. They underwent angiography and subsequent angioplasty for stenosis of 70% or greater. The exercise test was repeated post angioplasty. Careful screening for clinical endpoints and presence of ST segment depression and positive U-wave was done during exercise and the first 3 min of the recovery phase. RESULTS: Fifteen patients had isolated exercise induced U-wave and five had additional ST segment depression of > or =1 mm. Significant stenosis (>70% diameter reduction) of the circumflex artery was seen in 11 (55%) and of the right coronary artery in 9 (45%) patients. Coronary artery stenosis was reduced from 90+/-2% to 13+/-1% (mean+/-S.D.) P<0.001. On repeat of the exercise test U-wave and ST depression disappeared in all 20 individuals. Effort tolerance was improved after angioplasty for a mean duration of 3 min and 38 s, P<0.001. CONCLUSION: Exercise induced positive U-wave is an infrequent but specific marker of significant single coronary (circumflex or right) artery stenosis. It may lend itself to the detection of restenosis. PMID- 10841970 TI - Clinical and echocardiographic predictors of left atrial clot and spontaneous echo contrast in patients with severe rheumatic mitral stenosis: a prospective study in 200 patients by transesophageal echocardiography. AB - The objective of this study was to prospectively investigate various clinical and echocardiographic variables to predict the left atrial and left atrial appendage clot and spontaneous echo contrast in patients with severe rheumatic mitral stenosis. We studied 200 consecutive patients (112 males and 88 females; mean age 29.6+/-9.6 years). Left atrial clot and spontaneous echo contrast were present in 26 and 53.5% of cases, respectively. There were no significant differences in the mitral valve area, mean transmitral diastolic gradient and left ventricular ejection fraction between patients with and without clot. Patients with clot were older (34.4+/-11.4 vs. 28.2+/-8.5 years, P<0.001), had longer duration of symptoms (41. 4+/-36.0 vs. 28.8+/-22.9 months, P<0.001), more frequent atrial fibrillation and spontaneous echo contrast (69.2 vs. 16.9%, P<0. 00001 and 76.9 vs. 45.3%, P<0.00001, respectively) and larger left atrial area and diameter (41.0+/-12.7 vs. 29.9+/-7.4 cm(2), P<0.00001 and 53.9+/-8.3 vs. 47.6+/-7.4 mm, P<0.0001, respectively) as compared to patients without clot. Similarly patients with spontaneous echo contrast were older (31+/-10.4 vs. 27.8+/-8.3 years, P<0.01), had more frequent atrial fibrillation (48.6 vs. 9.7%, P<0.0001), left atrial clot (37.4 vs. 12.9%, P<0.0001), larger left atrial area and diameter (37.6+/-11.2 vs. 28.1+/-6.7 cm(2), P<0.00001 and 52.2+/-8.3 vs. 45.9+/-6.5 mm, P<0.00001, respectively) and smaller mitral valve area (0.77+/-0.14 vs. 0.84+/ 0.13 cm(2), P<0.01) as compared to patients without spontaneous echo contrast. There were no significant differences in the mean transmitral diastolic gradient and left ventricular ejection fraction. On multiple regression and discriminant function analysis, atrial fibrillation and left atrial area were independent predictors of left atrial clot formation. In a subgroup of patients with sinus rhythm, larger left atrial area and presence of spontaneous echo contrast were significantly associated with the presence of clot in left atrium and appendage. We conclude that in patients with severe mitral stenosis, the presence of atrial fibrillation and in the subgroup of the patients with sinus rhythm the presence of large left atrium (> or =40 cm(2)) and spontaneous echo contrast were associated with higher risk of clot formation in the left atrium and might be benefited by prophylactic anticoagulation. PMID- 10841971 TI - The roots of microbiology and the influence of Ferdinand Cohn on microbiology of the 19th century. AB - The beginning of modern microbiology can be traced back to the 1870s, and it was based on the development of new concepts that originated during the two preceding centuries on the role of microorganisms, new experimental methods, and discoveries in chemistry, physics, and evolutionary cell biology. The crucial progress was the isolation and growth on solid media of clone cultures arising from single cells and the demonstration that these pure cultures have specific, inheritable characteristics and metabolic capacities. The doctrine of the spontaneous generation of microorganisms, which stimulated research for a century, lost its role as an important concept. Microorganisms were discovered to be causative agents of infectious diseases and of specific metabolic processes. Microscopy techniques advanced studies on microorganisms. The discovery of sexuality and development in microorganisms and Darwin's theory of evolution contributed to the founding of microbiology as a science. Ferdinand Cohn (1828 1898), a pioneer in the developmental biology of lower plants, considerably promoted the taxonomy and physiology of bacteria, discovered the heat-resistant endospores of bacilli, and was active in applied microbiology. PMID- 10841972 TI - Molecular basis of antibiotic resistance and beta-lactamase inhibition by mechanism-based inactivators: perspectives and future directions. AB - Antibacterial chemotherapy is particularly striking in the family of penicillins and cephalosporins. Over 40 structurally different beta-lactam molecules are available in 73 formulations and the majority of them are currently prescribed for medical use in hospitals. beta-Lactams are well tolerated by humans with few side effects. They interact very specifically with their bacterial target, the D alanyl-D-alanine carboxypeptidase-transpeptidase usually referred to as DD peptidase. The outstanding number of beta-lactamases produced by bacteria represent a serious threat to the clinical utility of beta-lactams. The discovery of beta-lactamase inhibitors was thought to solve, in part, the problem of resistance. Unfortunately, bacteria have evolved new mechanisms of resistance to overcome the inhibitory effects of beta-lactamase inactivators. Here, we summarize the diversified mechanistic features of class A beta-lactamases interactions with mechanism-based inhibitors using available microbiological, kinetic and structural data for the prototype TEM beta-lactamases. A brief historical overview of the strategies developed to counteract beta-lactamases will be presented followed by a short description of the chemical events which lead to the inactivation of TEM beta-lactamase by inhibitors from different classes. Finally, an update on the clinical prevalence of natural and inhibitor resistant enzyme mutants, the total chemical synthesis to design and synthesize a new structure and produced a broad spectrum beta-lactamase inhibitor that mimics the beta-lactam ring, but does not contain it is discussed. PMID- 10841973 TI - Ecological significance of compatible solute accumulation by micro-organisms: from single cells to global climate. AB - The osmoadaptation of most micro-organisms involves the accumulation of K(+) ions and one or more of a restricted range of low molecular mass organic solutes, collectively termed 'compatible solutes'. These solutes are accumulated to high intracellular concentrations, in order to balance the osmotic pressure of the growth medium and maintain cell turgor pressure, which provides the driving force for cell extension growth. In this review, I discuss the alternative roles which compatible solutes may also play as intracellular reserves of carbon, energy and nitrogen, and as more general stress metabolites involved in protection of cells against other environmental stresses including heat, desiccation and freezing. Thus, the evolutionary selection for the accumulation of a specific compatible solute may not depend solely upon its function during osmoadaptation, but also upon the secondary benefits its accumulation provides, such as increased tolerance of other environmental stresses prevalent in the organism's niche or even anti-herbivory or dispersal functions in the case of dimethylsulfoniopropionate (DMSP). In the second part of the review, I discuss the ecological consequences of the release of compatible solutes to the environment, where they can provide sources of compatible solutes, carbon, nitrogen and energy for other members of the micro-flora. Finally, at the global scale the metabolism of specific compatible solutes (betaines and DMSP) in brackish water, marine and hypersaline environments may influence global climate, due to the production of the trace gases, methane and dimethylsulfide (DMS) and in the case of DMS, also couple the marine and terrestrial sulfur cycles. PMID- 10841974 TI - Sorting and function of peroxisomal membrane proteins. AB - Peroxisomes are subcellular organelles and are present in virtually all eukaryotic cells. Characteristic features of these organelles are their inducibility and their functional versatility. Their importance in the intermediary metabolism of cells is exemplified by the discovery of several inborn, fatal peroxisomal errors in man, the so-called peroxisomal disorders. Recent findings in research on peroxisome biogenesis and function have demonstrated that peroxisomal matrix proteins and peroxisomal membrane proteins (PMPs) follow separate pathways to reach their target organelle. This paper addresses the principles of PMP sorting and summarizes the current knowledge of the role of these proteins in organelle biogenesis and function. PMID- 10841975 TI - Periplasmic protein thiol:disulfide oxidoreductases of Escherichia coli. AB - Disulfide bond formation is part of the folding pathway for many periplasmic and outer membrane proteins that contain structural disulfide bonds. In Escherichia coli, a broad variety of periplasmic protein thiol:disulfide oxidoreductases have been identified in recent years, which substantially contribute to this pathway. Like the well-known cytoplasmic thioredoxins and glutaredoxins, these periplasmic protein thiol:disulfide oxidoreductases contain the conserved C-X-X-C motif in their active site. Most of them have a domain that displays the thioredoxin-like fold. In contrast to the cytoplasmic system, which consists exclusively of reducing proteins, the periplasmic oxidoreductases have either an oxidising, a reducing or an isomerisation activity. Apart from understanding their physiological role, it is of interest to learn how these proteins interact with their target molecules and how they are recycled as electron donors or acceptors. This review reflects the recently made efforts to elucidate the sources of oxidising and reducing power in the periplasm as well as the different properties of certain periplasmic protein thiol:disulfide oxidoreductases of E. coli. PMID- 10841976 TI - Redox factor-1 in muscle biopsies of patients with inclusion-body myositis. AB - To determine whether redox factor-1 (Ref-1) participates in the pathogenesis of inclusion-body myositis (IBM), we immunolocalized Ref-1 in muscle biopsies of IBM patients by light- and electron-microscopy. Approximately 70-80% of the IBM vacuolated muscle fibers had focal inclusions strongly immunoreactive for Ref-1. By immunoelectronmicroscopy, Ref-1 was localized to paired-helical filaments, 6 10 nm amyloid-like fibrils and amorphous material. Virtually all regenerating and necrotic muscle fibers in various muscle biopsies had diffusely strong Ref-1 immunoreactivity. At all neuromuscular junctions, postsynaptically there was strong Ref-1 immunoreactivity. Our study suggests that Ref-1 plays a role in IBM pathogenesis, and in other pathologic and normal processes of human muscle. PMID- 10841977 TI - Modulation of epileptiform discharges in the rat limbic system in vitro by noradrenergic agents. AB - Application of bicuculline (20 microm) in a horizontal slice preparation of the rat limbic system induced epileptiform discharges ('bursts') that spread from the hippocampus to the entorhinal cortex (EC) and the basolateral amygdala (BLA). These bursts were time locked, with the CA3-CA2 regions discharging first at a 0.1+/-0. 008Hz (n=30 slices) frequency, followed after 27+/-3 ms by the superficial layers of the EC and after 44.3+/-3 ms by the BLA. Application of 50 microM noradrenaline (NA) for 4 min reduced the burst frequency to 40% of its initial value. Pharmacological study of NA action on burst frequency revealed that it consisted of a beta adrenoreceptor-mediated increase and an alpha(2) adrenoreceptor-mediated decrease of epileptiform bursting frequency. PMID- 10841978 TI - Electroacupuncture potentiates the antinociceptive effect of intrathecal endomorphin-1 in the rat formalin test. AB - Endomorphin-1 is a novel endogenous mu-opioid peptide. In this study, we examined the effects of 2 Hz electroacupuncture in the rat tail flick test and the formalin test (a persistent noxious model). Moreover, we investigated if the electroacupuncture potentiated the effect of intrathecal endomorphin-1. The results demonstrated that electroacupuncture alone produced a significant antinociception in the tail flick test, but not in the formalin test, and that intrathecal endomorphin-1 dose-dependently suppressed the biphasic nociceptive behavior in the formalin test. Electroacupuncture enhanced the antinociceptive effect of intrathecal endomorphin-1 in the formalin test, resulting in a significant leftward shift in the dose-response curves for intrathecal endomorphin-1 antinociception. The enhanced effect was antagonized by intraperitoneal naltrexone. The study suggests that electroacupuncture may potentiate the intrathecal endomorphin-1 antinociception partially mediated by opioid receptors. PMID- 10841979 TI - Impairment of motor imagery in putamen lesions in humans. AB - Patients with putamen or cortical lesions participated in a first- and third person movement imagery task, each primarily engaging kinesthetic and visual imagery. The subjects were instructed to imagine themselves (first-person task) and a third party (third-person task) performing a sequence of three movements and to choose from a set of four photos the end posture resulting from the movements. The results demonstrated that, limb-specific imagery was impaired in both putamen and cortical lesions, in the first-, but not third-person task. Moreover, more than half of the errors made by cortical patients were with respect to the first movement, a finding consistent with motor cortex involvement in memory processes. Taken overall, the results provide evidence that the basal ganglia as well as cortical structures play an important role in the neural network mediating motor imagery. PMID- 10841980 TI - Activation of nuclear factor-kB in the spinal cord of experimental allergic encephalomyelitis. AB - The DNA-binding activity of nuclear factor (NF-kB) was found to be induced in the spinal cord of rats with experimental allergic encephalomyelitis (EAE), an animal model of multiple sclerosis (MS), from the onset of the disease. This activation of NF-kB persisted throughout the disease period and decreased thereafter in the recovery phase. Supershift analysis of NF-kB DNA-binding activity in nuclear extracts of spinal cords showed that RelA/p65 and p50 subunits but not c-Rel/p75, RelB/p68 and p52 subunits were involved in DNA binding. Pyrrolidine dithiocarbamate (PDTC), an inhibitor of NF-kB activation, markedly inhibited the in vivo activation of NF-kB in the spinal cord of EAE rats and attenuated the clinical symptoms of EAE. These studies suggest that activation of NF-kB plays an important role in the pathogenesis of EAE and inhibitors of NF-kB activation may have therapeutic importance in MS. PMID- 10841981 TI - Changes in electrophysiological properties of tibial motoneurones in the rat following 4 weeks of tetrodotoxin-induced paralysis. AB - In this study, we test the hypothesis that 4 weeks tetrodotoxin (TTX) paralysis altered the passive membrane properties of rat tibial motoneurones. Impulse activity along the sciatic nerve was blocked for 4 weeks using TTX delivered by an osmotic minipump to a Silastic cuff placed around the nerve. That portion of the sample exhibiting the 20% slowest After-hyperpolarization (AHP) decay time (AHPd), and which therefore included presumptive type S motoneurons, demonstrated responses (reduced AHPd, increased rheobase and rheobase voltage), which were not evident in the rest of the sample (presumptive fast motoneurons), in which an increased AHPd, in fact, was found. The results thus support the hypothesis that retrograde signals from inactive slow and fast muscle fibers have different effects on their innervating motoneurones. PMID- 10841983 TI - Optical recording of spatiotemporal activation of rat somatosensory and visual cortex in vitro. AB - A comparative analysis of spatiotemporal activation patterns of somatosensory and visual cortex was carried out in slice preparations using optical recording with voltage-sensitive dyes. Activity propagation velocities were found to be similar in both areas in all layers. Vertical propagation velocity is higher than horizontal propagation velocities. Differences between the two sensory areas exist in terms of horizontal activity spread, that is similar in extragranular layers but smaller in somatosensory than in visual cortex in layer IV. These results imply that despite the extensive similarities in the organization of sensory cortical areas, systematic areal variations in the horizontal cortical plane are present that may reflect adaptations needed for the processing of the corresponding sensory modality. PMID- 10841982 TI - The leucine (7)-to-proline (7) polymorphism in the signal peptide of neuropeptide Y is not associated with Alzheimer's disease or the link apolipoprotein E. AB - Both apolipoprotein E varepsilon4 allele (APOE varepsilon4) and neuropeptide Y (NPY) Pro(7)-variant have been reported to be associated with higher serum levels of total and LDL cholesterol. Since APOE varepsilon4 allele is also a major risk factor for the development of Alzheimer's disease (AD) and the genetic polymorphism of NPY has not previously been studied in dementing disorders, we have examined whether a novel polymorphism in a signal peptide of NPY gene is associated with AD alone or in combination with APOE varepsilon4. A total of 125 sporadic AD cases and 110 control individuals from Finland were genotyped for APOE and NPY genes using the polymerase chain reaction and restriction enzyme analysis. The APOE varepsilon4 allele frequency was significantly increased in the AD group compared with controls as expected. Instead, no significant differences were found between sporadic AD patients and controls either in the NPY genotype or allele frequencies or in combination with the APOE varepsilon4 allele. We conclude that APOE varepsilon4 allele represents a strong predictor of risk for AD. PMID- 10841984 TI - Prostaglandin E(2)-mediated sensitization of rat sensory neurons is not altered by nerve growth factor. AB - To ascertain whether chronic exposure to nerve growth factor (NGF) alters the responsiveness of sensory neurons to prostaglandin E(2) (PGE(2)), sensory neurons taken from adult rats were grown in culture in the presence or absence of NGF for 7 days. Neurons then were exposed to PGE(2) and release of immunoreactive calcitonin gene-related peptide (iCGRP) and production of immunoreactive cAMP (icAMP) were examined. Growing neurons in the presence of 250 ng/ml NGF increased the content and the release of iCGRP from sensory neurons. Independent of NGF treatment, exposure to 100 nM PGE(2) augmented capsaicin- or potassium-stimulated release of iCGRP by 1. 5-fold compared with cells not exposed to PGE(2). In a similar manner, NGF treatment did not alter the ability of PGE(2) to increase the content of icAMP. These data suggest that prostaglandin-induced sensitization of sensory neurons is not influenced by NGF. PMID- 10841985 TI - Lasting influence of repetitive transcranial magnetic stimulation on intracortical excitability in human subjects. AB - We studied the effects of a train of 30 pulses of repetitive transcranial magnetic stimulation (rTMS) at frequencies of 5 and 15 Hz and at an intensity of 120% of resting motor threshold on human motor cortex excitability. Intracortical inhibition (ICI) and intracortical facilitation (ICF) after rTMS were tested by a conditioning-test designed paired-pulse paradigm. After 15 Hz rTMS, ICI was significantly reduced for 3.2 min, ICF was enhanced for 1.5 min, muscle evoked potentials from single TMS were increased in the first 30 s only. After 5 Hz rTMS ICI was significantly reduced in the first 32 s. We conclude that high-frequency suprathreshold rTMS transiently suppresses the activity of the intracortical inhibitory circuitry and, with a different time course, increases the excitability of the excitatory circuitry. PMID- 10841986 TI - Effect of noradrenergic denervation of the amygdala upon recovery after sleep deprivation in the rat. AB - We previously showed that the noradrenergic locus coeruleus (NA-LC) was involved in the regulatory mechanisms of the paradoxical sleep rebound following a 10 h sleep deprivation by using a systemic injection of a specific neurotoxin, N-(2 chloroethyl)-N-ethyl-2-bromobenzylamine (DSP-4). Given that rebound mechanisms are mainly located in the forebrain, we planned to study the role of the forebrain structures receiving LC afferences. In this study we evaluated the involvement of noradrenergic afferences to the central nucleus of the amygdala in the sleep rebound by DSP-4 microinjections into the central nucleus of the rat amygdala. The results showed that during the first recovery day, the paradoxical sleep rebound is lower in DSP-4 treated rats (-67.28%). These findings indicate that the amygdala, through its NA afferents, contributes to the sleep rebound mechanisms. PMID- 10841988 TI - Motion sickness occurrence does not correlate with nystagmus characteristics. AB - The purpose of this study was to test the hypothesis whereby eye movements as such may be an important factor in the development of motion sickness (MS). The horizontal eye movements of 27 subjects were measured during earth vertical axis rotation (EVAR) and during off vertical axis rotation (OVAR). Two groups were set up, one including subjects who suffered severe MS during the test, and the other including subjects with no MS symptoms. We found no differences in nystagmus parameters (EVAR: gain and time constant; OVAR: eye velocity modulation and eye position modulation) between the MS and the non-MS groups. We can conclude that eye movements are not involved in the development of MS. PMID- 10841987 TI - Ginsenosides inhibit capsaicin-activated channel in rat sensory neurons. AB - Ginsenosides isolated from ginseng are biologically active components. In this study, whole-cell and inside-out configurations of patch clamp technique had been used to test the effect of ginsenosides on the capsaicin-activated channels in cultured small diameter sensory neurons of young rat. Ginsenosides (100 microg/ml) decreased the amplitude of capsaicin-activated currents by 78.2% in whole cell mode. Similarly, ginsenosides decreased capsaicin-activated single channel activities in a dose-dependent manner in inside-out patches. These results indicate that ginsenosides might directly block capsaicin-activated channels, resulting in attenuation of the currents in rat sensory neurons. PMID- 10841989 TI - Voltage- and gamma-aminobutyric acid-activated membrane currents in the human medulloblastoma cell line MHH-MED-3. AB - The whole-cell patch clamp technique was used to characterize voltage- and neurotransmitter-activated currents in the medulloblastoma cell line MHH-MED-3 and cells from tissue slices and primary cultures of two medulloblastoma biopsies. These preparations revealed similar electrophysiological properties. All tested cells displayed 4-aminopyridine-sensitive delayed rectifying K(+) currents, gamma-aminobutyric acid(A) receptor-mediated Cl(-) currents and most of them inward rectifier K(+) currents. Transient inward currents were mainly carried by low-voltage activated T-type Ca(2+) channels in MHH-MED-3 cells, and tetrodotoxin-sensitive Na(+) channels in cells from the primary culture. From these characteristics we conclude that medulloblastoma cells share physiological features with developing cerebellar granule cells at an immature stage. PMID- 10841990 TI - Neomycin blocks substance P-induced calcium entry in cultured rat spinal cord neurons. AB - Substance P (SP) plays an important role in sensitization of spinal cord neurons. In this study, we investigated SP-induced calcium activities in cultured rat spinal cord neurons with confocal laser scanning microscopy. Our results showed that SP increased [Ca(2+)](i) by calcium entry rather than release from intracellular calcium stores. Neomycin (100 microM), an antagonist of phosphatidylinositol 4,5-bisphosphate (PIP(2)), blocked SP-induced calcium entry. Ca(2+)-free medium induced capacitative entry, which was significantly potentiated by SP. As activation of SP receptor (NK-1) leads to production of inositol 1,4,5-trisphosphate (IP(3)) by PIP(2) turnover, the results indicates that SP-induced calcium entry and SP-potentiated capacitative calcium entry might be mediated or regulated by IP(3)/diacylglycerol (DG) pathway. PMID- 10841991 TI - Experimentally induced attenuation of neuropeptide-Y gene expression in transgenic mice increases mortality rate following seizures. AB - Previous experiments have reported increased seizure susceptibility in transgenic mice lacking normal neuropeptide-Y (NPY) gene expression (i.e. NPY 'knock-out' mice). A critical issue inherent in such experiments concerns the confounding of developmental influences of NPY and its neurotransmitter functions in the mature organism. The present experiments directly addressed this issue by studying seizure susceptibility in transgenic mice possessing an inducible antisense transcript that can be experimentally manipulated to attenuate NPY synthesis. NPY deficient and control mice were injected with kainic acid (40 mg/kg, i.p.) and several seizure-related behaviors were measured. Consistent with previously reported effects in NPY knock-out mice, significantly more NPY-deficient mice died within 24 h than control mice. In situ hybridization analyses confirmed a decrease in prepro-NPY gene expression in transgenic mice. The experiments support the hypothesis that the control of neural excitability is a prominent function of NPY. PMID- 10841992 TI - Full length alpha-synuclein is present in cerebrospinal fluid from Parkinson's disease and normal subjects. AB - Several clues suggest that alpha-synuclein, a presynaptic protein, plays a central role in the pathogenesis of idiopathic Parkinson's disease (PD). To search a peripheral marker of PD, we analyzed presence and amount of alpha synuclein in CSF from 12 PD patients and 10 neurologically normal subjects. The protein was extracted from CSF samples through immunoprecipitation and immunoblotting with different specific anti-alpha-synuclein antibodies. We identified a 19 kDa band that corresponds to monomeric alpha-synuclein, given its comigration with homologue human recombinant peptide as well as with the protein extracted from cerebral cortex of normal subjects. The amount of CSF 19 kDa alpha synuclein did not significantly vary in PD and normal cases. These findings have two implications: (a) full length alpha-synuclein is released by neurons in the extracellular space; (b) alpha-synuclein does not appear a peripheral marker of PD pathology. PMID- 10841993 TI - Spatial working memory performance after high-frequency repetitive transcranial magnetic stimulation of the left and right posterior parietal cortex in humans. AB - The effects of high-frequency repetitive transcranial magnetic stimulation (rTMS) at the left or right posterior parietal cortex were studied using a spatial working memory task. Eight subjects were stimulated over the P3 and P4 electrode site at 115% of the motor threshold (frequency 25 Hz, trains of 200 ms) during the 1000-ms delay of the spatial working memory task, or received sham stimulation. It was found that the reaction times were slower during right parietal rTMS than during left-parietal rTMS. No differences were found between the percentages correct responses. These results are in line with recent neuroimaging findings and data from patients with cerebral lesions, suggesting that the posterior parietal cortex is especially involved in spatial processing, and provide converging evidence for recent theories on hemispheric specialization. PMID- 10841994 TI - Co-distribution of NK2 tachykinin receptors and substance P in nerve endings of guinea-pig ileum. AB - The distribution of NK2 tachykinin receptors-immunoreactivity (NK2r-IR) in the guinea-pig ileum and the co-distribution of NK2r-IR with substance P immunoreactivity (SP)-IR were investigated. NK2r-IR was detected in varicose fibers of myenteric and submucous ganglia and nerve strands, in the longitudinal and circular muscle layers and at the deep muscular plexus (DMP). Except for the submucous plexus, some of the NK2r-IR varicose fibers were co-distributed with SP IR ones and quantitative analysis showed significant regional differences in the percentages of these fibers. These results demonstrate that presynaptic NK2 receptors are located at varicose fibers likely originating from motor neurons projecting to muscle layers and DMP, and from interneurons. Furthermore, the NK2r/SP-IR co-distribution suggests that some of these receptors are autoreceptors on SP nerve endings. PMID- 10841995 TI - Influence of cognitive strategies on the pattern of cortical activation during mental subtraction. A functional imaging study in human subjects. AB - Functional magnetic resonance imaging (fMRI) at 1.5 T was used to investigate the influence of cognitive strategies on cortical activation during mental calculation. Twenty-nine right-handed subjects performed a serial subtraction of prime numbers. Even though a common corpus of brain areas was activated during this mental calculation, differences appeared between subjects in function of their spontaneous cognitive strategy. In subjects using a so called verbal strategy (n=15), the main activation was located in the whole left dorsolateral frontal cortex with a little activation of the inferior parietal cortex. In subjects using a so called visual strategy (n=14), a bilateral activation in the prefrontal cortex and a high activation in the left inferior parietal cortex were observed. These results demonstrate that numbers are processed through a distributed network of cortical areas, the lateralization of which is clearly influenced by subject strategy. PMID- 10841996 TI - Field trial on the therapeutic efficacy of paromomycin on natural Cryptosporidium parvum infections in lambs. AB - The objective of this study was to evaluate the therapeutic efficacy of paromomycin against cryptosporidiosis in naturally infected lambs under field conditions. The 36 cross-bred neonatal lambs, 3-10 days old, were used. On the first day that lambs showed diarrhea (Day 1) they were randomly divided into three groups. The infected control group (14 lambs) remained unmedicated whereas the two other groups were orally medicated with paromomycin solution (Humatin((R)), Parke Davis, France): 12 lambs (Group A) at 100mg/kg per day for three consecutive days (Days 1-3) and 10 lambs (Group B) at 200mg/kg per day for two days (Days 1 and 2). Drug efficacy was assessed by evaluating the presence of diarrhea, oocyst shedding and weight gains from Days 1 to 23. The results show the efficacy of paromomycin in reducing both cryptosporidial oocyst output and severity of clinical signs. On Day 4, all unmedicated lambs remained infected and excreted large numbers of cryptosporidial oocysts (mean score: 2.5) whereas oocyst output had stopped in most medicated lambs (>60%) and low numbers of oocysts were excreted in the remaining lambs (mean score: 0.45 in Group A and 1 in Group B). Mean oocyst excretion was significantly reduced in medicated lambs from Days 2 to 5 (P<0.05). Treatment also reduced, but not completely prevented, clinical symptoms although diarrhea stopped in most medicated lambs just after drug withdrawal. The mean weight gains of Group A lambs were higher than that of unmedicated lambs throughout the study and statistically significant differences were found from Days 1 to 11 (1.99+/-0.81 versus 1.47+/-0.53) (P<0.05). By contrast, the growth rate of Group B lambs from Days 11 to 23 was impaired when compared with the two other groups (P<0.05) although no significant differences were found at the end of the study (Days 1-23). PMID- 10841997 TI - Serologic survey of Neospora caninum infection in a closed dairy cattle herd in Maryland: risk of serologic reactivity by production groups. AB - Prevalence of antibodies to Neospora caninum was determined in a cross-sectional consensus survey of 1029 bovines in a dairy herd with endemic Neospora-induced abortion. Sera were screened by indirect fluorescent antibody test (IFAT). The prevalence of N. caninum antibody in the IFAT was 17.9% in 107 neonates, 26.2% in 233 yearling heifers and steers, 39.07% in 218 mature heifers, and 26.9% in 465 milking cows. Serologic reactivity was associated with production grouping on the farm with the greatest risk of serologic reactivity appearing in the yearling and mature heifers. There was an increasing risk of serologic reactivity with increasing age only in the parity one and greater animals in the herd. Castrated males were at half the risk of similarly aged females of possessing antibodies to N. caninum. There was no clear relationship between the serologic status of dams and offspring. PMID- 10841998 TI - The role of CD4(+) or CD8(+) T cells in the protective immune response of BALB/c mice to Neospora caninum infection. AB - The role of CD4(+) or CD8(+) T cells in the immune response of BALB/c mice against Neospora caninum infection was examined by using anti-CD4 and/or anti-CD8 monoclonal antibodies (mAbs). Mice were intraperitoneally inoculated with anti CD4 and/or anti-CD8 mAbs before and after infection with N. caninum and observed for 30 days after infection. Most of the anti-CD4 mAb-treated mice and all of the anti-CD4 and anti-CD8 mAbs-treated mice died within 30 days post-infection (p.i.). In contrast, 100% of PBS-treated mice and 70% of anti-CD8 mAb-treated mice survived more than 30 days. When compared with phosphate-buffered saline (PBS)-treated mice, the weight of mice treated with mAbs tended to decrease. From these results CD4(+) T cells, but not CD8(+) T cells, have an important role for protection of mice against N. caninum infection. Serum antibody levels to N. caninum in infected-mice treated with anti-CD4 mAb or a mixture of anti-CD4 and anti-CD8 mAbs were lower than those in the infected mice treated with anti-CD8 mAb or PBS. The mice treated with anti-interferon-gamma (IFN-gamma) mAb produced high antibody levels to N. caninum, but all mice died within 18 days p.i. These results indicated that IFN-gamma is an important cytokine for protection against N. caninum infection at the early stage of infection. However, since CD4(+) T cells against N. caninum were essential to the production of specific antibody, these antibodies might have important roles in host protection at the later stage of infection. PMID- 10841999 TI - Prevalence and genotypic characterisation of Giardia in dairy calves from Western Australia and Western Canada. AB - In this study, the prevalence of Giardia duodenalis infections was determined in Western Canadian and Western Australian dairy calves. Faecal samples were collected from Holstein calves located on a commercial dairy near Lethbridge, Alta., Canada (N=28) and from calves located on two commercial dairies located near Perth, WA, Australia (N=36). Faecal samples were examined for the presence of Giardia cysts using sucrose gradient centrifugation, followed by immunofluoresence microscopy. DNA was then extracted from Giardia isolates obtained from positive samples. A PCR based method was employed to amplify and sequence a 292bp region of the 16S-rRNA gene. Genetic sequences obtained from Giardia isolates were compared to each other and to previously sequenced isolates. Following a single faecal sample, 58% of Western Australian calves and 57% of Western Canadian calves were positive for Giardia. Geometric mean cyst counts/g of faeces were 839 for Western Australian calves and 3475 for Western Canadian calves, but these values did not differ significantly. Genetic sequences were obtained from 10 calves from Western Canada, while six sequences were obtained from Western Australian calves. Of the Western Canadian isolates, eight aligned with the proposed 'Hoofed livestock' genotype. Of the five isolates obtained from Western Australian calves, four sequences were identical to the 'Hoofed livestock' genotype. Two isolates from the Western Canadian calves and one isolate from the Western Australian calves had the identical genetic sequence to the Genotype (Assemblage) A sequence, a common human genotype. The results of this study demonstrate, for the first time, that Giardia infections occur in Western Australian calves. Also, calves from different geographical locations appear to be primarily infected with a Giardia genotype unique to hoofed livestock. However, calves can shed Giardia cysts potentially infective for humans. Thus, Giardia infections should be considered important to Australian dairy producers, and infections in calves may pose a risk to public health regardless of geographical location. PMID- 10842000 TI - Serological responses of cattle after treatment and during natural re-infection with Fasciola hepatica, as measured with a dot-ELISA system. AB - The dot-ELISA reaction was used to study the dynamics of IgG titers in cattle naturally infected with Fasciola hepatica after anthelmintic treatment and during reinfection. Excretion/secretion products (ES) of the parasite were used as antigens for the dot-ELISA. IgG antibodies were no longer detectable by dot ELISA, 4-6 months after nine animals received the first of three weekly doses of triclabendazole (15 mg kg(-1)) and were then maintained on a pasture free of F. hepatica metacercariae. Six fluke-free cattle began shedding F. hepatica eggs 3-6 months after grazing a pasture contaminated with metacercariae of the parasite. A detectable increase in dot-ELISA IgG antibody levels was observed 2-4 weeks after natural reinfection by grazing a similar pasture contaminated with F. hepatica metacercariae. The usefulness of the dot-ELISA system to diagnose chronic infection by serology is complicated by previous treatment against the parasite. It is concluded that the ES antigens can be useful to detect early infection of cattle with F. hepatica in a dot-ELISA system PMID- 10842001 TI - Vaccination against Taenia solium cysticercosis in underfed rustic pigs of Mexico: roles of age, genetic background and antibody response. AB - Vaccination of pigs of mixed genetic make-up, raised as rustically as done in rural Mexico, resulted in effective protection to experimental challenge against Taenia solium cysticercosis. Maximum protection was achieved if pigs were immunized at 70 days of age. There was large variation of viable parasite load within vaccinated pigs and controls, which is suggestive of significant genetic factors influencing susceptibility, besides immunization. Our results strengthen the advisability of pig vaccination for control of T. solium cysticercosis, since it lowers the number of viable cysticerci capable of transforming into tapeworms. PMID- 10842002 TI - Host specificity of abomasal nematodes in free ranging alpine ruminants. AB - Abomasums from 641 alpine wild ruminants representing five different species (Cervus elaphus, Capreolus capreolus, Rupicapra rupicapra, Capra ibex, Ovis musimon) and from 19 domestic sheep (Ovis aries) from alpine areas were examined in order to investigate the host-specificity of abomasal helminths. Nine out of 20 helminth species were found in at least five different host species. A discriminant analysis was able to significantly discriminate the hosts on the basis of their helminth community composition with the exception of O. musimon and O. aries. Based on the correlation between each variable represented by helminth species with the most explanatory discriminant axis, it was possible to classify helminths into specialists and generalists. Specialists are represented by the dominant species in a community of an host species or family while generalists appear in the communities of many different hosts as intermediate species. Due to the pathogenic potential of some of these generalist species (i.e. Haemonchus contortus, Trichostrongylus axei) and their ability to adapt easily to the conditions found in several different hosts, they appear to be the most important from a sanitary point of view. PMID- 10842003 TI - Competence of Aedes albopictus and Culex quinquefasciatus as vector of Dirofilaria immitis after blood meal with different microfilarial density. AB - Aedes albopictus and Culex quinquefasciatus were fed canine blood with different microfilarial density of Dirofilaria immitis ranging from 2500 to 25,000 mff/ml. Larval development in these two mosquito species did not differ significantly. Although C. quinquefasciatus ingested more microfilariae, the number of larvae which developed in A. albopictus was invariably greater than in C. quinquefasciatus. Mortality of the engorged A. albopictus was significantly greater than that of C. quinquefasciatus, and higher microfilarial density raised the mortality in both species. The vector efficiency index of A. albopictus was greater than C. quinquefasciatus at all microfilarial densities, but its survival time was much reduced. Thus, dogs with low microfilarial density are implicated as the main source for the transmission of D. immitis from dogs to mosquitoes. PMID- 10842004 TI - Expected effects of reducing Oestrus ovis L. mature larval weight on adult populations. AB - In order to estimate the effects of eventual reductions in larval weight (LW) of Oestrus ovis L. as a measure of control, the correlation between mature LW and adult fly length (AL) in laboratory specimens (n=150) was calculated. The regression equation AL=5.62+10.65LW (r(2)=0.76) was obtained. This equation was then applied to estimate the mature LW of wild larvipositing females (n=51) to predict the minimum mature LW at which fly viability would be compromised. The critical weight, 0.28 g (standard error limits 0. 235, 0.323), was obtained from a small fly measuring 8.6mm in length. Data from 383 mature third instars were used to estimate, by statistical analysis, the expected effects of decreasing the mature LW on subsequent fly population size. A considerable mean reduction (38%) in adult populations might be achieved by a 40% reduction of mature LW, but this eventual reduction may be temporary due to the high reproductive rate in this species. Sex differences in mature LW and fly size are also reported. PMID- 10842005 TI - Isolation of Neospora caninum from the brain of a naturally infected adult dairy cow. AB - Neospora caninum was isolated from the brain of a 2-year-old dairy cow that had aborted confirmed N. caninum-infected fetuses on two occasions. The cow had an indirect fluorescent antibody titer of 1:1600 to N. caninum. The cow was killed 24 days after its second abortion and the brain was bioassayed for N. caninum in nude mice. Multifocal areas of perivascular cuffing and glial nodules were observed in the cerebrum and mesencephalon of the cow, but N. caninum was not identified in histological sections of the brain. All three nude mice inoculated with brain homogenate of the cow, developed emaciation and paralysis. Microscopical examination of the nude mice revealed systemic N. caninum infection with demonstrable tachyzoites in various organs. The parasites isolated from fresh mouse brain were transferred successfully into Vero cell cultures. PCR procedure on the purified tachyzoites obtained from the Vero cell cultures amplified the specific DNA sequence for N. caninum. PMID- 10842006 TI - Responses of neurons in the cat posteromedial lateral suprasylvian cortex to moving texture patterns. AB - The posteromedial lateral suprasylvian cortex represents a point of convergence between the geniculostriate and extrageniculostriate visual pathways. Given its purported role in motion analysis and the conflicting reports regarding the texture sensitivity of this area, we have investigated the response properties of cells in PMLS to moving texture patterns ("visual noise"). In contrast to previous reports, we have found that a large majority of cells (80.1%) responds to the motion of a texture pattern with sustained discharges. In general, responses to noise were more broadly tuned for direction compared to gratings; however, direction selectivity appeared more pronounced in response to noise. The majority of cells was selective for drift velocity of the noise pattern (mean optimal velocity: 26.7 degrees /s). Velocity tuning was comparable to that of its principal thalamic input, the lateral posterior pulvinar nucleus. In general, responsiveness of cells in the posteromedial lateral suprasylvian cortex increased with increasing texture element size, although some units were tuned to smaller element sizes than the largest presented. Finally, the magnitude of these noise responses was dependent on the area of the visual field stimulated. In general, a stimulus corresponding to roughly twice the size of the receptive field was required to elicit an equivalent half-maximal response to that for gratings. The results of this study indicate that the majority of cells in the posteromedial lateral suprasylvian cortex can be driven by the motion of a fine texture field, and highlight the importance of this area in motion analysis. PMID- 10842007 TI - Spatial properties and direction selectivity of single neurons in area 21b of the cat. AB - The receptive field properties of single units were assessed in area 21b of the cat visual cortex. Visual cells in this area were binocular and showed relatively large receptive fields. Most cells were strongly sensitive to the direction of drifting gratings. The mean value of the half-widths of the direction tuning curves (32 degrees ) suggests broader direction tunings than are typically found in other visual areas. The spatial frequency tuning functions were either band pass or low-pass. Cells responded optimally to low spatial frequencies (mean =0.08c/deg) and also showed low spatial resolution (mean =0.29c/deg.). The estimated values of spatial bandwidths (mean=2.2 octaves) suggest that area 21b cells act as relatively good spatial filters. Although some cells exhibited a low contrast threshold, most cells began to respond at intermediate or high contrast values (mean threshold =15.5%). Temporal frequency tuning functions were mostly band-pass and usually broad (mean temporal bandwidth=3.3 octaves). Cells were found that responded optimally to various temporal frequencies (mean optimal temporal frequency=3.2Hz), although the majority preferred a temporal frequency below 4Hz.These results suggest that visual properties (receptive fields sizes, spatial resolution and orientation/direction selectivity) of cells in area 21b differ from those of cells previously observed in the adjoining area 21a. These differences provide evidence in support of functional distinction between these two visual areas. PMID- 10842008 TI - Limited collateralization of neurons in the rat prefrontal cortex that project to the nucleus accumbens. AB - The specificity and selectiveness of a neuronal message depends in part on the number of recipient neurons that simultaneously receive this message. Hence, projections involved in higher order cognitive processes might be expected to exhibit a lower degree of collateralization than projections that mediate more basic brain functions. This study sought to determine the degree to which neurons projecting from the prefrontal cortex to the nucleus accumbens collateralize to major cortical and subcortical regions: the contralateral prefrontal cortex, the basolateral amygdala or the ventral tegmental area. Fluoro-Gold and cholera toxin b were used to label prefrontal cortex neurons that project to these targets, and the proportion of neurons singly and dually labeled by immunofluorescence for these tracers was determined. The prefrontal cortex neurons projecting to these regions exhibited a partially complementary laminar distribution. Furthermore, of the neurons projecting to the nucleus accumbens, 13% sent a collateralized projection to the contralateral prefrontal cortex, 7% collateralized to the basolateral amygdala, and 3% sent a branched projection to the ventral tegmental area. No differences were observed in the degree of collateralization of neurons in superficial versus deep layers.Thus, the degree of collateralization of corticoaccumbens neurons was overall limited, but significantly greater to a cortical target than to subcortical regions. These branching patterns provide anatomical substrates for temporal and spatial coordination of activity in limbic circuits. PMID- 10842009 TI - Cortical projections from the suprasylvian gyrus to the reticular thalamic nucleus in the cat. AB - The cat's suprasylvian gyrus was injected iontophoretically with either 4% wheat germ agglutinin-horseradish peroxidase, 4% dextran-fluororuby or 4% dextran biotin. The locations of labelled fibres, presumed terminals and cell bodies were determined with the aid of a camera lucida attachment and computer aided stereometry. Cells from the crown of the suprasylvian gyrus project to the dorsal most portion of the rostral half of the reticular nucleus. The region or 'sector' is distinct, albeit with some overlap, from the visual sector of the reticular nucleus defined by projections from adjacent extrastriate visual cortices. The projection from the suprasylvian gyrus to the reticular nucleus has a rough topography such that the caudal areas project to the more caudal aspects of the sector and rostral areas project to the more rostral areas of the reticular nucleus. There is a large degree of overlap of rostrocaudal projections from the suprasylvian gyrus within the sector, however, the projections originating from rostral sites are situated in a more ventral location compared to the projection originating from the caudal suprasylvian gyrus. Analysis of the distribution of biotin labelled presumptive terminals did not support the notion of 'slabs' or regional variation in terminal density across the mediolateral thickness of the reticular nucleus. In addition, a number of presumptive terminals were found within the internal capsule which coincided with the position of retrogradely labelled cells in the internal capsule following thalamic injections and appears to be part of the perireticular nucleus. The results suggest that the reticular nucleus may be segregated into sectors connected with modality specific cortical areas (e.g. striate and extrastriate visual areas) and nonspecific sectors connected with polymodal (e.g. area 7) cortical regions. The reticular nucleus and its connections with the suprasylvian gyrus may form an important link in binding eye movements to sensory integrative process through visuomotor and auditory thalamic connections. PMID- 10842010 TI - The supramammillo-hippocampal and supramammillo-septal glutamatergic/aspartatergic projections in the rat: a combined [3H]D-aspartate autoradiographic and immunohistochemical study. AB - It is well established that the supramammillary nucleus plays a critical role in hippocampal theta rhythm generation/regulation by its direct and indirect (via the septal complex) connections to the hippocampus. Previous morphological and electrophysiological studies indicate that both the supramammillo-hippocampal and supramammillo-septal efferents contain excitatory transmitter. To test the validity of this assumption, transmitter specific retrograde tracer experiments were performed. [3H]D-aspartate was injected into different locations of the hippocampus (granular and supragranular layers of the dentate gyrus and CA2 and CA3a areas of the Ammon's horn) and septal complex (medial septum and the area between the medial and lateral septum) that are known targets of the supramammillary projection. Consecutive vibratome sections prepared from the entire length of the posterior hypothalamus, including the supramammillary area, were immunostained for calretinin, tyrosine hydroxylase, or calbindin, and further processed for autoradiography. Radiolabeled, radiolabeled plus calretinin containing, and calretinin-immunoreactive neurons were plotted at six different oro-caudal levels of the supramammillary area. The results demonstrated that following both hippocampal and septal injection of the tracer, the majority of the retrogradely radiolabeled (glutamatergic/aspartatergic) cells are immunoreactive for calretinin. However, non-radiolabeled calretinin-containing neurons and radiolabeled calretinin-immunonegative cells were also seen, albeit at a much lower density. These observations clearly indicate the presence of glutamatergic/aspartatergic projections to both the hippocampus and septal complex. It may be assumed that this transmitter could play a role in hippocampal theta rhythm generation/regulation. PMID- 10842011 TI - Cholinergic modulation of synaptic transmission and plasticity in entorhinal cortex and hippocampus of the rat. AB - Effects of cholinergic agents on synaptic transmission and plasticity were examined in entorhinal cortex and hippocampus. Bath application of carbachol (0.25-0.75 microM) induced transient depression of field potential responses in all cases tested (24/24 in layer III of medial entorhinal cortex slices and 24/24 in CA1 of hippocampal slices; 11.0+/-1.9% and 7.8+/-2.5%, respectively) and long lasting potentiation in some cases (4/24 in entorhinal cortex and 12/24 in hippocampus; 33.7+/-3.7% and 32.1+/-9.9%, respectively, in successful cases). Carbachol (0.5 microM) induced transient depression, but not long-lasting potentiation, of N-methyl-D-aspartate receptor-mediated responses in entorhinal cortex. At 5 microM, carbachol induced transient depression only (55. 9+/-4.7% in entorhinal cortex and 41.4+/-2.9% in hippocampus), which was blocked by atropine. Paired-pulse facilitation was not altered during carbachol-induced potentiation but enhanced during carbachol-induced depression. These results suggest that the underlying mechanisms of carbachol-induced depression and potentiation are decreased transmitter release and selective enhancement of non-N-methyl-D aspartate receptor-mediated responses, respectively. Long-term potentiation could be induced in the presence of 10 microM atropine by theta burst stimulation. The magnitude was significantly lower (15.2+/-5.2%, n=9) compared with control (37.2+/-6.1%, n=8) in entorhinal cortex, however. These results demonstrate similar, but not identical, cholinergic modulation of synaptic transmission and plasticity in entorhinal cortex and hippocampus. PMID- 10842012 TI - Repeated electroconvulsive shock promotes the sprouting of serotonergic axons in the lesioned rat hippocampus. AB - This study reports the effect of repeated electroconvulsive shock on the sprouting of 5-hydroxytryptamine neurons in the partly lesioned rat dorsal hippocampus. We have adopted a 5-hydroxytryptamine homotypic collateral sprouting model to examine whether electroconvulsive shock administration altered the rate of 5-hydroxytryptamine axonal reinnervation of the dorsal hippocampus. The 5 hydroxytryptamine innervation of hippocampus originates from the median raphe via the cingulum bundle and the fimbria-fornix. Lesioning of the cingulum bundle has previously been shown to cause sprouting of intact 5-hydroxytryptamine afferents originating from the unharmed fimbria-fornix. Rats were unilaterally injected with the 5-hydroxytryptamine neurotoxin, 5,7-dihydroxytryptamine, into the right cingulum bundle and 5-hydroxytryptamine immunoreactivity in the dorsal hippocampus was investigated 1, 3, 6 and 12weeks after the injection. The lowest level of 5-hydroxytryptamine-immunoreactivity in the hippocampus was detected at three weeks after the lesion. At six weeks, 5-hydroxytryptamine immunoreactive fibres started to reappear, and at 12weeks the level of 5-hydroxytryptamine immunoreactivity was similar to that observed on the unlesioned side. Based on this time-course, six weeks was chosen as the time-point to investigate the action of a course of repeated electroconvulsive shock administrations. Repeated electroconvulsive shock (five shocks over 10days) doubled the number of sprouting 5-hydroxytryptamine-immunoreactive fibres and significantly increased levels of the 5-hydroxytryptamine metabolite, 5-hydroxyindoleacetic acid. The present data provide the first direct evidence that electroconvulsive shock enhances 5 hydroxytryptamine axon sprouting in the partly lesioned hippocampus. This is an effect which may contribute to the therapeutic effect of electroconvulsive therapy in major depression. PMID- 10842013 TI - Synaptic loss is accompanied by an increase in synaptic area in the dentate gyrus of aged human apolipoprotein E4 transgenic mice. AB - To investigate the relationship between the three isoforms of apolipoprotein E (E2, E3 and E4) and the integrity of the synaptic circuitry in the dentate gyrus of the hippocampus, we have estimated the synapse per neuron ratio and mean apposition zone area per synapse at the electron microscope level in the dentate gyrus of apolipoprotein E knockout and human apolipoprotein E transgenic mice aged six to 24months. During ageing, only in human apolipoprotein E4 mice was there a decrease in synapse per neuron ratio, accompanied by an increase in synaptic size. When these mice were compared with human apolipoprotein E2, apolipoprotein E knockout and wild-type mice at old age, they displayed the lowest synapse per neuron ratio, but similar apposition zone area. In contrast, as in our previous study, aged apolipoprotein E knockout mice did not show any sign of synaptic degeneration. The functional consequences of such morphological changes remain to be determined. However, if such age-related loss of synapses occurred in the brain of Alzheimer apolipoprotein E4 patients, they might be additive to pathological processes and could contribute to greater cognitive impairment. PMID- 10842014 TI - Specific induction of early growth response gene 1 in the lateral nucleus of the amygdala following contextual fear conditioning in rats. AB - Although the amygdala is known to be crucial for fear conditioning, little is known about the molecular and cellular mechanisms in the amygdala that are important for fear conditioning. One possible mechanism may be the activation of immediate-early genes, which function as regulatory factors of transcriptional processes. To investigate whether one of the major immediate-early gene families is involved in the learning and memory of fear, we examined the effects of fear conditioning on the expression of the four members of the early growth response (EGR) gene family, EGR-1, EGR-2, EGR-3, and EGR-4. Image analysis of in situ hybridization of messenger RNA of the four family members was performed in the amygdala, hippocampus, and neocortex 15, 30 and 60min following one-trial contextual fear conditioning. Rats were either handled, placed within the testing context without receiving the footshock, and received a footshock immediately upon placement within the context, or received a footshock after a 3-min delay (delayed-shock). Of the four groups, only the delayed-shock group exhibited a fear response (freezing). EGR-1 messenger RNA expression in the dorsolateral part of the lateral amygdaloid nucleus was significantly greater in the delayed-shock group compared with the other groups 15 and 30min following the conditioning. The increased expression of EGR-1 was specifically localized to the lateral nucleus of the amygdala; expression in the hippocampus and cortex was not increased by fear conditioning. In contrast, the expression of EGR-2, EGR-3, and EGR-4 messenger RNA was not increased in the amygdala, hippocampus or cortex following fear conditioning. In addition, following a retention test conducted 24h after fear conditioning, no increases were found in the expression of EGR-1 messenger RNA expression in the amygdala, hippocampus or cortex. The results demonstrate that of the four genes of the EGR family of transcription-regulatory factors, only EGR-1 messenger RNA in the dorsolateral portion of the lateral nucleus of the amygdala was specifically increased with contextual fear conditioning. It is suggested that EGR-1 plays a functional role during learning, but not retrieval, of contextual fear within the lateral nucleus of the amygdala. PMID- 10842016 TI - Prion protein deposition and abnormal synaptic protein expression in the cerebellum in Creutzfeldt-Jakob disease. AB - Prion protein (PrP(C)) is a cell membrane-anchored glycoprotein, which is replaced by a pathogenic protease-resistant, beta-sheet-containing isoform (PrP(CJD) or PrP(SC)) in human and animal prion encephalopathies, including sporadic Creutzfeldt-Jakob disease. Cell fractionation methods show that PrP(C) localizes in presynaptic membrane-enriched fractions. Following infection, abnormal PrP accumulates in nerve cell processes and synaptic regions. The present study examines the possible correlation between abnormal PrP deposition and the expression of synaptic proteins controlling neurotransmission in the cerebellum of six 129 Met/Met sporadic cases of Creutzfeldt-Jakob disease. Aggregates of protease-resistant PrP-positive granules, reminiscent of cerebellar glomeruli, were found in the granular cell layer, whereas fine punctate PrP immunoreactive deposits occurred in the molecular layer. Small numbers of diffuse, irregular plaque-like PrP deposits in the molecular and granular cell layers were present in every case. The somas of Purkinje cells, and stellate, basket and Golgi neurons, were not immunostained. PrP-immunoreactive fibres were found in the album of the cerebellum and hilus of the dentate nucleus. Punctate PrP deposition decorated the neuropil of the dentate nucleus and the surface of dentate neurons. Synaptic protein expression was examined with synaptophysin, synapsin-1, synaptosomal-associated protein of 25,000 mol. wt, syntaxin-1 and Rab3a immunohistochemistry. Reduced synaptophysin, synapsin-1, synaptosomal associated protein of 25,000 mol. wt, syntaxin-1 and Rab3a immunoreactivity was noted in the granular cell layer in every case, but reduced expression was inconstant in the molecular layer. Synaptophysin accumulated in axon torpedoes, thus indicating abnormal axon transport. Expression of synaptic proteins was relatively preserved in the dentate nucleus, although synaptophysin immunohistochemistry disclosed large coarse pericellular terminals in Creutzfeldt Jakob disease, instead of the fine granular terminals in control cases, around the soma of dentate neurons. Finally, Rab3a accumulated in the cytoplasm of Purkinje cells, thus suggesting major anomalies in Rab3a transport. These observations demonstrate, for the first time, abnormal expression of crucial synaptic proteins in the cerebellum of cases with Creutzfeldt-Jakob disease. However, abnormal PrP deposition is not proportional to the degree of reduction of synaptic protein expression in the different layers of the cerebellar cortex and in the dentate nucleus. Therefore, it remains to be elucidated how abnormal PrP impacts on the metabolism of proteins linked to exocytosis and neurotransmission, and how abnormal PrP deposition results in eventual synaptic loss. PMID- 10842015 TI - Electrophysiological characteristics of substantia nigra neurons in organotypic cultures: spontaneous and evoked activities. AB - Morphological and electrophysiological characteristics of dopaminergic and non dopaminergic neurons in the substantia nigra and their postsynaptic responses to stimulation of the tegmental pedunculopontine nucleus were studied in rat organotypic triple cultures. These cultures consisted of the subthalamic nucleus explant, ventral mesencephalic explant, inclusive of the substantia nigra and the mesopontine tegmentum explant, inclusive of the tegmental pedunculopontine nucleus, prepared from one- to two-day-old rats. Intracellular sharp and whole cell recordings were obtained from three- to eight-week-old organotypic cultures. Recorded neurons were identified as dopaminergic and non-dopaminergic neurons with tyrosine hydroxylase immunohistochemistry. Dopaminergic neurons had long duration action potentials, prominent afterhyperpolarization, time-dependent inward and outward rectification and strong frequency adaptation. Spontaneous firing patterns varied from regular, irregular to burst firing. Non-dopaminergic neurons had short duration action potentials, in general no rectifying currents, and maintained high firing frequencies. Spontaneous firing patterns in these neurons were irregular or burst firing. Morphological analysis of the recorded neurons labeled with neurobiotin revealed that non-dopaminergic neurons had more extensive arborization of higher-order dendrites than dopaminergic neurons. Dopaminergic and non-dopaminergic neurons receive glutamatergic and cholinergic excitatory inputs from the tegmental pedunculopontine nucleus. These results indicate that morphological and electrophysiological characteristics of substantia nigra neurons in the organotypic culture are generally similar to those reported in in vitro slice and in vivo studies. However, spontaneous activities of dopamine neurons observed in the organotypic culture preparation more closely resemble those in in vivo preparation compared to in vitro preparation. PMID- 10842017 TI - The effects of neurotrophin-3 and brain-derived neurotrophic factor on cerebellar granule cell movement and neurite extension in vitro. AB - Migration of the granule cells is a major stage of cerebellar maturation. Granule cells express neurotrophins and their receptors; however, their role in cell migration has not been defined. In this study we investigated the effects of exogenous neurotrophins on the movement and neurite extension of granule cells from glial-free cerebellar cell reaggregates in vitro. Our results provide direct evidence that neurotrophin-3 and brain-derived neurotrophic factor differentially affect the granule cells. Neurotrophin-3 significantly affected granule cell movements by decreasing the migration index (the ratio of the number of cells that moved further than half the neurite length) and the speed of cell soma movement, but did not affect neurite length or growth cone migration. In contrast, brain-derived neurotrophic factor and neurotrophin-4 acted on growing neurites and growth cones by significantly increasing neurite length and the speed of growth cone migration, but had no effect either on the migration index or on the speed of the cell soma movement. The results suggest that neurotrophins differentially affect neurite extension and the movements of cerebellar granule cells. PMID- 10842018 TI - Transforming growth factor-beta 2 is anterogradely and retrogradely transported in motoneurons and up-regulated after nerve injury. AB - The survival of motoneurons is dependent on them receiving continual trophic support from muscle fibres and various other cell types. Numerous putative survival factors have been identified and a set of criteria established by which these candidates can be assessed. These criteria include the need for the factor and its receptors to be in appropriate locations and for the factor or its second message to be retrogradely transported. In this paper, we demonstrate that a multifunctional cytokine, transforming growth factor-beta 2, appears to meet these criteria. The locations of the transforming growth factor-beta 2 and its receptors in the neuromuscular system were determined by reverse transcriptase polymerase chain reaction and immunohistochemistry. Motoneurons were shown to synthesize the three proteins involved in transforming growth factor-beta 2 signalling (types I and II transforming growth factor-beta receptor and betaglycan) and to transport them anterogradely, where they were inserted into the axonal membrane and nerve terminal. Transforming growth factor-beta 2 was detected in the synaptic portions of muscle fibres, motoneurons and in injured nerves, indicating that motoneurons may be exposed to multiple and potentially redundant sources of transforming growth factor-beta 2. Double-ligation experiments were used to demonstrate that motoneurons transport transforming growth factor-beta 2 up and down their axons. The anterograde transport of both transforming growth factor-beta 2 and its receptors, coupled with the fact that most of a motoneuron's mitochondria are located in the axon, raises the issue of whether the repression of the initiation of apoptosis is restricted to the cell body or occurs along the entire length of a neuron. PMID- 10842019 TI - Spinal cyclooxygenase-2 is involved in development of allodynia after nerve injury in rats. AB - Increased spinal cyclooxygenase activity is associated with nociception induced by tissue inflammation. In the present study, we examined the changes of cyclooxygenase-1 and cyclooxygenase-2 protein expression in several regions of the CNS associated with pain perception, and the role of spinal cyclooxygenase activity in the development of allodynia following nerve injury. Allodynia was induced by ligation of the left L5 and L6 spinal nerves in rats. Using western blot analysis, we found that the cyclooxygenase-2 protein levels in the dorsal spinal cord and thalamus (but not in the ventral spinal cord, cingulate cortex and locus coeruleus) increased significantly one day after nerve ligation, compared with those in the sham animals. The cyclooxygenase-2 protein levels in the above tissues were similar in nerve-injured and sham animals three and 14 days after surgery. In contrast, cyclooxygenase-1 protein was not detectable in any of the neural tissues examined one, three, and 14 days after nerve injury. In the behavioral experiments, we observed that intrathecal injection of 100microg of indomethacin immediately or one day after nerve ligation attenuated the development of tactile allodynia. However, intrathecal injection of indomethacin had no effect on established allodynia two weeks after nerve injury.Collectively, our results suggest that cyclooxygenase-2 is preferentially up-regulated in the dorsal spinal cord and thalamus in response to nerve injury in rats. Spinal cyclooxygenase-2 probably plays an important role in the early development, but not in the maintenance, of tactile allodynia caused by the nerve injury in this rat model of neuropathic pain. PMID- 10842020 TI - Reduction by naloxone of lipopolysaccharide-induced neurotoxicity in mouse cortical neuron-glia co-cultures. AB - An inflammatory response in the CNS mediated by activation of microglia is a key event in the early stages of the development of neurodegenerative diseases. Using mouse cortical mixed glia cultures, we have previously demonstrated that the bacterial endotoxin lipopolysaccharide induces the activation of microglia and the production of proinflammatory factors. Naloxone, an opioid receptor antagonist, inhibits the lipopolysaccharide-induced activation of microglia and the production of proinflammatory factors. Using neuron-glia co-cultures, we extended our study to determine if naloxone has a neuroprotective effect against lipopolysaccharide-induced neuronal damage and analysed the underlying mechanism of action for its potential neuroprotective effect. Pretreatment of cultures with naloxone (1 microM) followed by treatment with lipopolysaccharide significantly inhibited the lipopolysaccharide-induced production of nitric oxide and the release of tumor necrosis factor-alpha, and significantly reduced the lipopolysaccharide-induced damage to neurons. More importantly, both naloxone and its opioid-receptor ineffective enantiomer (+)-naloxone were equally effective in inhibiting the lipopolysaccharide-induced generation of proinflammatory factors and the activation of microglia, as well as in the protection of neurons. These results indicate that the neuroprotective effect of naloxone is mediated by its inhibition of microglial activity and may be unrelated to its binding to the classical opioid receptors. PMID- 10842021 TI - High concentrations of extracellular potassium enhance bacterial endotoxin lipopolysaccharide-induced neurotoxicity in glia-neuron mixed cultures. AB - A sudden increase in extracellular potassium ions (K(+)) often occurs in cerebral ischemia and after brain trauma. This increase of extracellular K(+) constitutes the basis for spreading depression across the cerebral cortex, resulting in the expansion of neuronal death after ischemic and traumatic brain injuries. Besides spreading depression, it has become clear that cerebral inflammation also is a key factor contributing to secondary brain injury in acute neurological disorders. Experiments to validate the relationship between elevated levels of extracellular K(+) and inflammation have not been studied. This study aims to elucidate the roles of high concentrations of extracellular K(+) in bacterial endotoxin lipopolysaccharide-induced production of inflammatory factors. Increased concentration of KCl in the medium (20mM) significantly enhanced neurotoxicity by lipopolysaccharide in glia-neuron mixed cultures. To delineate the underlying mechanisms of increased neurotoxicity, the effects of high extracellular K(+) were examined by using mixed glial cultures. KCl at 20mM significantly enhanced nitrite, an index for nitric oxide, production by about twofold, and was pronounced from 24 to 48h, depending on the concentration of KCl. Besides nitric oxide production of tumor necrosis factor-alpha was also enhanced. The augmentative effects of high KCl on the production of inflammatory factors were probably due to the further activation of microglia, since high KCl also enhanced the production of tumor necrosis factor-alpha in microglia-enriched cultures. The increased production of nitrite by high K(+) was eliminated through use of a K(+)-blocker. Taken together, the results show that increases of extracellular K(+) concentrations in spreading depression augment lipopolysaccharide-elicited neurotoxicity, because production of inflammatory factors such as nitric oxide and tumor necrosis factor-alpha are potentiated. Since spreading depression and cerebral inflammation are important in acute neurological disorders, the present results suggest a biochemical mechanism: elevated extracellular K(+) concentrations augment glial inflammatory responses, and thus the neurotoxicity. PMID- 10842022 TI - Disruption of actin cytoskeleton in cultured rat astrocytes suppresses ATP- and bradykinin-induced [Ca(2+)](i) oscillations by reducing the coupling efficiency between Ca(2+) release, capacitative Ca(2+) entry, and store refilling. AB - Oscillations of [Ca(2+)](i) which are believed to be important in regulation of cellular behaviour or gene expression, require Ca(2+) entry via capacitative Ca(2+) influx for store refilling. However, the mediator between Ca(2+) store content and activation of Ca(2+) influx is still elusive. There is also controversy about the role of the actin cytoskeleton in this coupling. Therefore, the importance of an intact actin cytoskeleton on ATP- and bradykinin-elicited Ca(2+) signalling was investigated in cultured rat astrocytes by treatment with cytochalasin D which changes the morphology of the cells from an extended to a rounded shape. Cytochalasin D-treated astrocytes were unable, upon prolonged stimulation with the P2Y receptor agonist ATP, to generate oscillations of [Ca(2+)](i) which are, however, seen in 54% of untreated control cells. In cytochalasin D-treated cells, the amplitude of the initial Ca(2+) response was reduced mainly by disturbing the Ca(2+) influx, and, moreover, the total Ca(2+) pool which is sensitive to thapsigargin or cyclopiazonic acid was diminished.Thus, disruption of the cytoskeleton blocks agonist-elicited [Ca(2+)](i) oscillations apparently by reducing the coupling efficiency between intracellular Ca(2+) stores and capacitative Ca(2+) entry. PMID- 10842023 TI - GABA(B)-mediated action in the frog olfactory bulb makes odor responses more salient. AB - In the olfactory bulb, GABA(B) receptors are selectively located in the glomerular layer. A current hypothesis is that GABAergic inhibition mediated through these receptors would be, at least partly, presynaptic and would exerted by decreasing the release of the olfactory receptor neuron excitatory neurotransmitter. Here, we assessed, in the frog, the in vivo action of baclofen, a GABA(B) agonist, on single-unit mitral cell activity in response to odors. Local application of baclofen in the glomerular region of the olfactory bulb was shown to drastically affect mitral cell spontaneous activity, since they became totally silent. Moreover, under baclofen, mitral cells still responded to odors and still specified odor concentration increases through their temporal response patterns. The pharmacological specificity of the GABA(B) agonist action was confirmed by showing that saclofen, a GABA(B) antagonist, partly prevented the inhibitory action of baclofen and restored the initial rate of mitral cell spontaneous activity. The results show that GABA(B)-mimicked inhibition suppressed mitral cell spontaneous activity while odor responses were maintained. This suggests that olfactory receptor neurons partly drive spontaneous mitral cell activity. Moreover, the effect of GABA(B)-mediated inhibition was seen to be very close to that described previously for dopamine D(2) receptor-mediated inhibition. In conclusion, we propose that these two inhibitory mechanisms would offer the possibility to reduce or suppress mitral cell spontaneous activity so as to make their responses to odor especially salient. PMID- 10842024 TI - Impaired behavioral suppression by light in metabotropic glutamate receptor subtype 6-deficient mice. AB - The metabotropic glutamate receptor subtype 6 is localized on the dendrites of ON bipolar cells in mammalian retina, and is responsible for synaptic transmission from photoreceptors to ON bipolar cells. We have previously provided electrophysiological evidence that metabotropic glutmate receptor subtype 6 deficient mice have an impairment in the ON visual pathway. In this study, we compared, between metabotropic glutamate receptor subtype 6-deficient (n=9) and wild-type mice (n=7), their daily wheel-running activity in constant dark and light-dark cycle environments. There was no difference in their free-running rhythmicity in a constant dark environment nor in their ability to entrain their active/rest phase to the phase-shifted light-dark cycle environment, indicating that the circadian system in mutant mice was functioning normally. However, the wheel-running activity was suppressed immediately after light onset of the light dark cycle in wild-type mice (suppressive effect), whereas that of mutant mice was prolonged for several hours in spite of light onset (very weak suppressive effect). The suppression of activity in wild-type mice is a "masking effect" of the endogenous circadian rhythm in response to light stimuli. The results indicate that the failure of mutant mice to suppress their activity upon light onset is not due to abnormality in their circadian system, but to their lack of response to light stimuli. This study clearly demonstrates that the dysfunction of the ON visual pathway in metabotropic glutamate receptor subtype 6-deficient mice impairs their behavioral responsiveness to light and yet preserves their circadian system. PMID- 10842025 TI - Fatigue effects in the cat gastrocnemius during frequency-modulated efferent stimulation. AB - Effects of low- and high-frequency fatigue were studied on muscle dynamics in isometric conditions of the cat gastrocnemius. Fatiguing sessions consisted of 25 28 repetitions of the standard tests that included an 18-s interval of continuous frequency-modulated stimulation preceded and followed by single stimuli evoking twitch contractions. The rate of the continuous part was changed in accordance with a symmetrical double-trapezoidal signal, including three successive phases of constant rate at 10, 40 and 10s(-1); between these phases, each lasting for 4s, the rate changed linearly within a 2-s interval. The following modes of muscle activation were applied: (i) stimulation of single filaments constituting approximately one-fifth to one-seventh of the total cross-section of the L(7) and S(1) ventral roots; (ii) the distributed stimulation of five similar filaments; and (iii) direct stimulation of muscle through bipolar wire electrodes. A relative drop in tension, the fatigue index, expressed as the ratio at the end of a fatigue session over its value at the beginning of the test, was used to quantify fatigue effects. The fatigue indices during low-rate stimulation were 0.56+/-0.03 (mean+/-S.D.) at the first phase and 0. 64+/-0.02 at the third phase, while during high-rate stimulation this parameter was only 0.32+/-0.02. The high rate stimulation noticeably increased the mean tension during low-rate stimulation; the ratio between the reactions at the third and the first phases could be as much as two to three times greater than that at the beginning of the fatigue session. It was demonstrated that the potentiation was connected with after-effects of the rate-tension hysteresis. The hysteresis decreased with fatigue, the fatigue index for the rate-tension loop areas ranging from 0.39 to 0.52 (0.45+/-0. 05, mean+/-S.D.). The fatigue processes developed more quickly and intensively in the previously fatigued muscles: the obtained fatigue indices were 0.73+/-0.05 and 0.70+/-0.10 at the first and third phases, and 0.62+/-0.06 (mean+/-S.D.) at the second phase of stimulation, respectively. In the cases of distributed and direct stimulation applied to muscles in a fresh state, fatigue dynamics did not differ significantly from those observed during single-filament stimulation. In experiments with distributed stimulation applied to previously fatigued muscles, a powerful depression of the high-rate components was registered in several cases, which seemed to be connected with depressive effects at the level of nerve-muscle synaptic transmission. The effects of low- and high frequency fatigue were studied in isometric conditions of muscle contraction. In addition to the well-known differentiation between low- and high-frequency fatigue effects, the complex pattern of efferent stimulation used allowed us to identify additional fatigue-related changes in the rate-tension hysteresis. This hysteresis seems to be one of the possible mechanisms directed to compensate for low-frequency fatigue in the muscle contraction. PMID- 10842026 TI - Fatigue-related changes in electomyogram activity of the cat gastrocnemius during frequency-modulated efferent stimulation. AB - Changes in the compound muscle action potentials of cat gastrocnemius muscle were studied during low- and high-frequency fatigue. Fatiguing session consisted of 25 28 repetitions of the standard single fatigue tests (1.5min interval between the tests) that included the part of continuous frequency-modulated stimulation preceded and followed by single stimuli evoking twitch contractions in the muscle. The rate of the continuous part was changed in accordance with symmetrical double-trapezoidal signal, including three successive phases of constant rate at 10, 40 and 10s(-1); between these phases of 4s duration the rate changed linearly within a 2s interval. During fatigue relative changes in compound muscle action potential waves were usually smaller than changes in tension. Within the same fatigue procedure applied to a fresh muscle, the drop in tension was as much as 35% for high-rate stimulation and 59-71% for low-rate stimulation, whereas the decrease of the peak-to-peak compound muscle action potential waves amplitudes did not exceed 10-20%. Compound muscle action potential waves underwent the most pronounced depression during high-rate stimulation, the decrease proceeding during the following phase of low-rate stimulation. The tension changes during long-lasting activation were different for low- and high-frequency fatigue, with more pronounced depression during low rate stimulation. As a rule, compound muscle action potential waves changes followed opposite patterns. Compound muscle action potential waves progressively split up, which was probably associated with a continuous slowing of the action potentials in the most fatigable motor units and the subsequent disappearance of the reactions at least in part of the motor units. Hysteresis effects in muscle contraction seem to be able, at least in part, to compensate for some of the depressive effects appearing during conduction of action potentials in muscle fibres. Changes in the compound muscle action potentials were studied during development of the muscle fatigue. These changes showed pronounced dependency on stimulation rate allowing differentiating effects of low- and high-frequency stimulation of the efferents supplying muscle under study. At the same time the fatigue-related changes in the action potentials were noticeably smaller than changes in tension, thus supporting existing concepts in the field arguing that fatigue effects are mainly connected with corresponding activity-dependent changes in muscle contraction machinery. PMID- 10842027 TI - Segmental analysis for cocaine and metabolites by HPLC in hair of suspected drug overdose cases. AB - Hair samples of eight postmortem cases were analyzed in segments of 1 to 3 cm for cocaine, benzoylecgonine and cocaethylene. Samples were prepared for analysis by digestion in 0.1 M HCl and subsequent extraction with mixed-mode solid-phase extraction columns. Measurement was made by reversed-phase, narrow-bore HPLC and fluorescence detection using two laboratory-made internal standards. The concentrations were in the region of 0.29-316 ng/mg of hair for cocaine, 0.43-141 ng/mg of hair for benzoylecgonine and 0.93-1.83 ng/mg of hair for cocaethylene. All eight investigated cases had cocaine-positive segments. In six of the cases, all segments were positive, suggesting regular cocaine use and two showed in between negative segments indicating an interruption or a change of the abuse intensity. The results showed a second, remarkable observation, i.e. enormous concentration differences (factor >150) for both cocaine and benzoylecgonine between the different subjects. Furthermore, interindividual cocaine/benzoylecgonine ratios ranged from 0.02 to 8.43. We believe these observations could in part be attributed to both some of the still existing limitations in the analytical approach(es), especially the mandatory hair washing steps, and in our still too limited knowledge of the hair incorporation processes. Nevertheless, in some cases, segmental analysis proved to be an important tool to distinguish, together with postmortem examination, deadly chronic abuse from single acute drug overdosage. PMID- 10842028 TI - A study on ten short tandem repeat systems: African immigrant and Spanish population data. AB - This work presents the results obtained from a genetic-population study for the D1S1656 system in the population of Southwest Spain (Huelva, Cadiz and Sevilla), Spaniards of Caucasian origin from North Africa (Ceuta), as well as in the black Central West African and Moroccan immigrant populations in Spain. The results of a study of the autochtonous population of the Canary Islands (n=138), and immigrant Central West African populations in Spain (n=132), obtained for nine short tandem repeat (STR) loci (D3S1358, VWA, FGA, D8S1179, D21S11, D18S51, D5S818, D13S317, D7S820), as well as the amelogenin locus, all contained in Profiler Plus (Perkin-Elmer) PCR amplification kits, are also presented. Except for the FGA and VWA data on immigrant Central West African populations in Spain, no deviations from the Hardy-Weinberg equilibrium were detected. PMID- 10842029 TI - Age estimation in small children: reference values based on counts of deciduous teeth in Finns. AB - The eruption of teeth in the mouth is suitable for age estimations during the period when teeth are actively emerging, in the deciduous dentition phase approximately from the age of 6 months to 2.5 years. Estimations of age can be performed simply by counting the number of teeth in the mouth. Reliability of the estimates depends on the reference data available and each population group should preferably have its own standards. In the present study timing of eruption of successive deciduous teeth was studied longitudinally in 129 Finns. The dates of clinical eruption of deciduous teeth were recorded by mothers and checked by dentists. In 40 of the 129 children emergence ages of at the most the four last teeth were based only on semiannual registrations performed by dentists. The main purpose was to provide normal timetables of tooth eruption in small children in forms that are practical in estimations of dental age. No sexual dimorphism existed in the timing of clinical eruption of successive deciduous teeth. The mean age corresponding to the presence of one tooth in the mouth was 7.1 months (S.D.=1.78) and that corresponding to tooth count 19 was 27.8 months (S.D.=3.99). If the chronological age is known, the presented distributions and means with variations make it possible to estimate the degree of advancement or delay in a child's dental development. If the age of the child is not known, the mean and median ages can be used for estimations of chronological age. However, estimations of age should not be based only on tooth counts because of marked variation also within this homogeneous group. PMID- 10842030 TI - Immunohistochemical study of tyrosine phosphorylation signaling in the involuted thymus. AB - Thymic involution has been reported to be an important parameter of the degree and duration of child abuse. In the present study, we assessed the status of tyrosine phosphorylation signaling, which is known to play a key role in the physiological function of the thymus, in involuted thymuses of abused children through immunohistological studies performed with anti-phosphotyrosine antibodies. We found that tyrosine-phosphorylated proteins were present in high amounts in Hassall's corpuscles (HC) in the medulla of control thymuses. In involuted thymuses of abused children, expression of tyrosine-phosphorylated proteins was reduced with accompanying morphological changes of HC, such as reduction in size or calcification. These findings lead us to the suggestion that tyrosine phosphorylation signaling is reduced in involuted thymuses of abused children and that reduction of the signaling may be associated with morphological changes of HC as observed in involuted thymuses of abused children. In order to certify the suggestion, we investigated expression of tyrosine-phosphorylated proteins in involuted thymuses of stressed rats as well as in control thymuses. Immunohistochemistry revealed that tyrosine-phosphorylated proteins were expressed in control thymuses, more abundantly in the medulla, and reduced remarkably in involuted thymuses of stressed rats. Further, immunoblot analysis also showed that expression of phosphotyrosine-containing proteins was reduced in thymus extracts of involuted thymuses of stressed rats, thus supporting the suggestion. Our results also raise the possibility that components of tyrosine phosphorylation signaling could be a molecular marker for thymic involution. PMID- 10842031 TI - Delimitation of the time of death by immunohistochemical detection of thyroglobulin. AB - To improve the possibilities to delimit the time of death after longer laytime it was examined if this is possible by immunohistochemical detection of thyroglobulin. The results show that in our examination material the colloid and the follicular cells of the thyroid glands of up to 5-day-old corpses produce a positive immunoreaction towards thyroglobulin in all cases whereas none of the corpses older than 13 days show such a reaction. This means that in case of a negative immunoreaction the time of death can be assumed to lie more than 6 days before the autopsy. The fact that a negative immunoreaction occurs consistently after 13 days leads to the conclusion that when thyroglobulin has been stained in a specimen, the death of the respective person must lie a maximum of 12 days earlier, whereby these time-limits may change in considerably different surrounding conditions. PMID- 10842032 TI - The statistical variability of blood alcohol concentration measurements in drink driving cases. AB - Like many other places in the world, Hong Kong has drink-driving legislation which prohibits a driver from having in his blood alcohol exceeding a prescribed limit while in control of a motor vehicle. The accuracy of measuring this alcohol concentration is obviously of prime concern as an erroneous result can avert the administration of justice. The common practice is to deduct all errors from the measured value and compare the deducted value with the prescribed limit, so that the benefit of all errors of the measurement is given to the driver. It is therefore important for any laboratory responsible for measuring blood alcohol concentrations to identify and quantify all errors associated with the measurement. The present study examined 900 blood alcohol determinations carried out by the Hong Kong Government Laboratory (HKGL) on cases of suspected drink driving. The determinations were performed by 5 different analysts with two different sets of instruments during 1995-1997. Statistical analysis indicated that the instruments had no bearing on the random error or variability and that even though analyst was a significant factor on variability, the deviation from the mean so caused was only 0.3% and of no practical significance. When the systematic error introduced by the tolerance limits of the certified alcohol standards (purchased from the Laboratory of Government Chemists, UK) was taken into account, the total uncertainty (random plus systematic errors) of an alcohol determination at 99.5% confidence level was found to be 4%. It is recommended that laboratories engaged in blood alcohol determination should adopt similar statistical treatment of their analytical results to find out the error and to ensure that the results are independent of analyst and instrument used. PMID- 10842033 TI - The challenge of identification following the tragedy of the Solar Temple (Cheiry/Salvan, Switzerland). AB - On October 5, 1994, 48 members of the Sect of the Solar Temple were found dead at two different locations in Switzerland: 23 victims in Cheiry and 25 victims in Salvan. Our Institute was commissioned to solve the forensic problems presented by this tragedy. Our goals were to establish the time of death, determine its causes, help elucidate the surrounding circumstances and identify the victims. This work presented us with the following challenges. This catastrophe was of an 'open' type: there was no list of 'passengers'; the victims were of five different nationalities and many had just arrived in Switzerland to participate in this event; family ties were very complex within this group; half of the victims were burned and sometimes charred; the exceptionally intense media converage of the story put a lot of pressure on the investigators and our Institute. In spite of these difficulties, all the victims were positively identified within 1 month. In the present report, we describe the steps realized to progress in our work. A special section describes our relationship with the journalists and their invaluable help in our investigations. The importance of being prepared for such an event is discussed. PMID- 10842045 TI - Automatic breast region extraction from digital mammograms for PACS and telemammography applications. AB - High spatial resolution results in very large digital mammogram file sizes. For telemammography, and picture archiving and communication systems, the large file issue introduces technical difficulties in image transmission, storage, and display. We propose extracting the breast region from the mammogram to reduce the image file size. The challenge is on how to faithfully extract breast regions from digital mammograms generated from different types of acquisition systems that contain various imaged compositions. We report an algorithm to automatically identify the orientation of breast region and extract the breast region from mammograms. Breast regions extracted from full-field digital mammograms reduce file sizes by three to five folds. PMID- 10842046 TI - Comparison of texture analysis methods for the characterization of coronary plaques in intravascular ultrasound images. AB - Intravascular Ultrasound (IVUS) is a diagnostic imaging technique that provides tomographic visualization of coronary arteries. The aim of this study was to evaluate five texture analysis techniques and determine their ability to distinguish between plaque lesions of different composition. Using histological correlation, regions of calcified, fibrous, and necrotic core plaque were chosen from 27 coronary plaques. First-order statistics, Haralick's method, Laws' texture energy method, the neighborhood gray-tone difference matrix method, and texture spectrum features were examined using discriminant analysis. Self validation indicated that Haralick's method yielded the most accurate results, with resubstitution and cross-validation error rates of 0.00 and 14.76%, respectively. Further optimization gave error rates of 6.67%, using only two discriminating features, IDM and entropy. PMID- 10842047 TI - The use of three-dimensional computed tomographic angiography in the accurate diagnosis of internal carotid artery aneurysms: degree for expression of posterior communicating and anterior choroidal arteries. AB - We have improved the scan protocol of three-dimensional computed tomographic angiography (3D-CTA) to maximize the accuracy of examining unruptured cerebral aneurysms. Sixty-seven cases of suspected internal carotid-posterior communicating (Pcom) and internal carotid-anterior choroidal (Ant Cho) artery aneurysms were evaluated as to the opacification of arteries. In the 46 cases of suspected fetal-type Pcom and Ant Cho aneurysms, the arteries were opacified from its origin to the periphery in all cases. A final diagnosis of the presence of an aneurysm was made possible by complete opacification. A definite diagnosis was impossible to make in only ten cases having adult-type Pcom. In conclusion, 3D CTA, using our own scan protocol, was proven to be quite useful for making a more accurate diagnosis. PMID- 10842048 TI - Three-dimensional skeletonization for computer-assisted treatment planning in radiosurgery. AB - This paper describes a new algorithm for skeletonization of two- (2D) and three dimensional (3D) objects based on ridge extraction. Ridges are formed when grassfire fronts collapse during grassfire propagation and they correspond to the locus of skeleton. The iso-distance contours/surfaces of a distance map are analogous to the grassfire fronts. They are locally smooth everywhere except at ridge locations. The new skeletonization algorithm extracts these ridge points based on local curvature measurement and is rotational invariant. It requires one scan of the image for curvature detection in any dimension, and is much faster than thinning methods. Connectivity checks are not required and the algorithm is extensible to higher dimensions. Our 3D skeletonization method is used in a novel algorithm to guide computerized planning of radiosurgical treatment of brain tumors. PMID- 10842049 TI - A large pseudolesion in the left lobe of the liver caused by inferior vena caval obstruction secondary to metastatic retroperitoneal lymphadenopathy. AB - A rare case with a large pseudolesion in the left lobe of the liver observed on early phase of incremental dynamic computed tomography (CT) caused by inferior vena caval obstruction is presented. Decreased portal perfusion due to increased volume of systemic venous inflow into the left lobe via paraumbilical venous system may be its underlying hemodynamic change. The etiology of this pseudolesion was successfully confirmed by conventional CT with intravenous contrast medium injection via superficial vein of lower extremity. PMID- 10842050 TI - Pseudolesion in segment IV of the liver with focal fatty deposition caused by the parabiliary venous drainage. AB - A case with a pseudolesion associated with focal fatty deposition in segment IV of the liver observed on conventional CT and CT during arterial portography caused by the parabiliary venous drainage is presented. Close observation of the common hepatic angiography was helpful to recognize this unusual vessel as a cause of this pseudolesion. Selective catheterization of the posterior superior pancreaticoduodenal artery and CT during its venous phase confirmed the etiology of the pseudolesion. PMID- 10842051 TI - Echinococcus of the pulmonary artery: CT, MRI and MRA findings. AB - The location of echinococcal cysts inside pulmonary artery is extremely rare. The cause is usually rupture of intracardiac cysts. We report a case of a 67-year-old patient with known lung hydatid disease whose main clinical presentation was dyspnea. The patient did not have any surgery in the past. The importance of the present case lies in the demonstration of the MR angiographic findings. PMID- 10842052 TI - Germ cell tumors of the central nervous system originating from non-pineal regions: CT and MR features. AB - CT and MR findings were retrospectively reviewed in 12 patients with germ cell tumors originating from the non-pineal regions. Cystic or necrotic components were seen in 6 patients. Of 4 germinomas, 2 showed mixed density on the CT. The MR signal intensity of the tumor was non-specific. Of 8 germinomas, 4 were inhomogeneously enhanced on postcontrast CT and T1-weighted MR images. CT and MR features of germinomas originating from non-pineal regions frequently differ from those of germinomas originating from the pineal region. The mass of the tumor often appeared cystic and inhomogeneously enhanced following contrast infusion. PMID- 10842053 TI - Unusual MRI findings in rhombencephalosynapsis. AB - Rhombencephalosynapsis is mainly characterized by fusion of the cerebellar hemispheres and vermian agenesis. We report severe cerebellar hypoplasia, pachygyria of the cerebrum, an isolated (trapped) fourth ventricle, and facial hemangioma in association with this condition. Our findings suggested that the frequency of rhombencephalosynapsis appears to be higher than previously thought (approximately 0.13%), and it should not be misinterpreted as Chiari II malformation. PMID- 10842054 TI - Su(H)-independent activity of hairless during mechano-sensory organ formation in Drosophila. AB - Formation of mechano-sensory organs in Drosophila involves the selection of neural precursor cells (SOPs) mediated by the classical Notch pathway in the process of lateral inhibition. Here we show that the subsequent cell type specifications rely on distinct subsets of Notch signaling components. Whereas E(spl) bHLH genes implement SOP selection, they are not required for later decisions. Most remarkably, the Notch signal transducer Su(H) is essential to determine outer but not inner cell fates. In contrast, the Notch antagonist Hairless, thought to act upon Su(H), influences strongly the entire cell lineage demonstrating that it functions through targets other than Su(H) within the inner lineage. Thereby, Hairless and numb may have partly redundant activities. This suggests that Notch-dependent binary cell fate specifications involve different sets of mediators depending on the cell type considered. PMID- 10842055 TI - Platelet-derived growth factor A modulates limb chondrogenesis both in vivo and in vitro. AB - Cartilage formation in the chick limb follows rapid proliferation, condensation and differentiation of limb mesenchyme. The control of these early events is poorly understood. Platelet-derived growth factor receptor alpha (PDGFR-alpha) is present throughout the mesenchyme of early chick limb buds, while its ligand, PDGF-A, is expressed in the surrounding epithelium. PDGFR-alpha is down-regulated in areas that will not give rise to cartilage and is then lost from cartilage forming areas after they begin to differentiate. PDGF-A increases chondrogenesis in micromass cultures of stage-20-24 limb buds, but not stage 25, where it inhibits chondrogenesis. Ectopic PDGF-A in the chick wing can lead to either a localized increase in cartilage formation, or an inhibition. Inhibition of PDGF signalling in the chick limb results in the loss of cartilage. These data demonstrate that PDGF-A functions to promote chondrogenesis at early stages of limb development and suggest that it inhibits chondrogenesis at later stages. PMID- 10842056 TI - Multiple functions of fibroblast growth factor-8 (FGF-8) in chick eye development. AB - Fibroblast growth factor-8 (FGF-8) is an important signaling molecule in the generation and patterning of the midbrain, tooth, and limb. In this study we show that it is also involved in eye development. In the chick, Fgf-8 transcripts first appear in the distal optic vesicle when it contacts the head ectoderm. Subsequently Fgf-8 expression increases and becomes localized to the central area of the presumptive neural retina (NR) only. Application of FGF-8 has two main effects on the eye. First, it converts presumptive retinal pigment epithelium (RPE) into NR. This is apparent by the failure to express Bmp-7 and Mitf (a marker gene for the RPE) in the outer layer of the optic cup, coupled with the induction of NR genes, such as Rx, Sgx-1 and Fgf-8 itself. The induced retina displays the typical multilayered cytoarchitecture and expresses late neuronal differentiation markers such as synaptotagmin and islet-1. The second effect of FGF-8 exposure is the induction of both lens formation and lens fiber differentiation. This is apparent by the expression of a lens specific marker, L Maf, and by morphological changes of lens cells. These results suggest that FGF-8 plays a role in the initiation and differentiation of neural retina and lens. PMID- 10842057 TI - Phenotypic effects in Xenopus and zebrafish suggest that one-eyed pinhead functions as antagonist of BMP signalling. AB - Zebrafish one-eyed pinhead (oep) is essential for embryonic axis and dorsal midline formation by promoting Nodal signalling and is thought to act as a permissive factor. Here we describe that oep elicits profound phenotypic effects when overexpressed in Xenopus and zebrafish. In Xenopus, wild-type oep inhibits mesoderm induction, disrupts axis formation and neuralizes animal caps. A secreted Oep dorsoanteriorizes and neuralizes Xenopus embryos indicative of BMP inhibition. In zebrafish, misexpression of smad1 in oep mutant embryos also reveals an interaction of oep with BMP signalling. Furthermore, the phenotypic effect of nodal overexpression can be rescued by coexpression of oep both in Xenopus and zebrafish. Taken together, our results support an interaction between oep and nodal but they suggest also (1) that the role of oep in Nodal signalling may include negative as well as positive regulation, (2) that oep is able to function in an active fashion and (3) that oep exerts a regulatory effect on the BMP signalling pathway. PMID- 10842058 TI - Proximal distal axis formation in the Drosophila leg: distinct functions of teashirt and homothorax in the proximal leg. AB - The proximal distal axis of the Drosophila leg is patterned by expression of a number of transcription factors in discrete domains along the axis. The homeodomain protein Homothorax and the zinc-finger protein Teashirt are broadly coexpressed in the presumptive body wall and proximal leg segments. Homothorax has been implicated in forming a boundary between proximal and distal segments of the leg. We present evidence that Teashirt is required for the formation of proximal leg segments, but has no role in boundary formation. PMID- 10842059 TI - Cactin, a conserved protein that interacts with the Drosophila IkappaB protein cactus and modulates its function. AB - Rel transcription factors function in flies and vertebrates in immunity and development. Although Rel proteins regulate diverse processes, the control of their function is conserved. In a two-hybrid screen for additional components of the pathway using the Drosophila I-kappaB protein Cactus as a bait, we isolated a novel coiled-coil protein with N-terminal Arg-Asp (RD)- like motifs that we call Cactin. Like the other components of this pathway, Cactin is evolutionarily conserved. Over-expression of cactin in a cactus(A2) heterozygous background results in the enhancement of the cactus phenotype. Both the embryonic lethality and ventralization are strongly increased, suggesting that cactin functions in the Rel pathway controlling the formation of dorsal-ventral embryonic polarity. PMID- 10842060 TI - Keratinocyte expression of transgenic hepatocyte growth factor affects melanocyte development, leading to dermal melanocytosis. AB - Using the epidermis-specific cytokeratin 14 promoter to deliver HGF exclusively from epidermal keratinocytes, we have examined the potential of hepatocyte growth factor (HGF) secreted from the normal environment to control morphogenesis. The transgenic mice displayed a significant increase of the number of melanocytes and their precursors in embryos starting not later than 16.5 dpc, and then after birth an explosive increase of dermal melanocytes started within 1 week, and these melanocytes were maintained throughout the entire life of the mice. Thus, HGF acts as a paracrine agent to promote survival, proliferation and differentiation of melanocyte precursors in vivo, and eventually causes melanocytosis. Loss of E-cadherin expression in dermal melanocyte precursors suggests that HGF caused dermal localization of melanocytes and their precursors by down-regulation of E-cadherin molecules. PMID- 10842061 TI - Ectopic expression of luteinizing hormone-releasing hormone and peripherin in the respiratory epithelium of mice lacking transcription factor AP-2alpha. AB - The vertebrate transcription factor activator protein-2 (AP-2alpha) is involved in craniofacial morphogenesis. In the nasal placode AP-2alpha expression delineates presumptive respiratory epithelia from olfactory epithelia, with AP 2alpha expression restricted to the anterior region of the respiratory epithelium (absent from the olfactory epithelium) at later stages. To address the role AP 2alpha plays in differentiation of cell groups in the nasal placode, the spatiotemporal expression pattern of four markers normally associated with olfactory epithelial structures was analyzed in mice lacking AP-2alpha. These markers were the intermediate filament protein peripherin, the neuropeptide luteinizing hormone-releasing hormone (LHRH), the neural cell adhesion molecule (NCAM) and the olfactory transcription factor Olf-1. Development of cells expressing these markers was similar in both genotypes until embryonic day 12.5 (E12.5), indicating that the main olfactory epithelium and olfactory pit formation was normal. At E13.5 in mutant mice, ectopic LHRH neurons and peripherin axons were detected in respiratory epithelial areas, areas devoid of Olf-1 and NCAM staining. Over the next few days, an increase in total nasal LHRH neurons occurred. The increase in nasal LHRH neurons could be accounted for by LHRH neurons arising and migrating out of respiratory epithelial regions on peripherin-positive fibers. These results indicate that AP-2alpha is not essential for the separation of the olfactory and respiratory epithelium from the nasal placode and is consistent with AP-2alpha preventing recapitulation of developmental programs within the respiratory epithelium that lead to expression of LHRH and peripherin phenotypes. PMID- 10842062 TI - Cell behaviour of Drosophila fat cadherin mutations in wing development. AB - We have studied several cell behaviour parameters of mutant alleles of fat (ft) in Drosophila imaginal wing disc development. Mutant imaginal discs continue growing in larvae delayed in pupariation and can reach sizes of several times those of wild-type. Their growth is, however, basically allometric. Homozygous ft cells grow faster than their twin cells in clones and generate larger territories, albeit delimited by normal clonal restrictions. Moreover, ft cells in clones tend to grow towards wing proximal regions. These behaviours can be related with failures in cell adhesiveness and cell recognition. Double mutant combinations with alleles of other genes, e.g. of the Epidermal growth factor receptor (DER) pathway, modify ft clonal phenotypes, indicating that adhesiveness is modulated by intercellular signalling. Mutant ft cells show, in addition, smaller cell sizes during proliferation and abnormal cuticular differentiation, which reflect cell membrane and cytoskeleton anomalies, which are not modulated by the DER pathway. PMID- 10842063 TI - Dissecting GHRH- and pituitary adenylate cyclase activating polypeptide-mediated signalling in Xenopus. AB - The highly conserved neuropeptide pituitary adenylate cyclase activating polypeptide (PACAP) has been implicated in a broad variety of physiological processes. The PACAP precursor protein gives rise to three different peptides, the cryptic peptide, GHRH, and PACAP, respectively, and here we dissect their functional properties using Xenopus as model system. PACAP and GHRH but not the cryptic peptide directly neuralize animal caps. In contrast to GHRH, the neuralizing effect mediated by PACAP is independent of the PKA pathway. Moreover, PACAP but not GHRH behaves like a BMP-4 antagonist. Blastocoel injection of PACAP 38 but not of the closely related peptides PACAP-27 and VIP leads to strong anteriorization of the injected embryos suggesting the possible involvement of a novel PACAP-preferring receptor. PMID- 10842064 TI - Indian hedgehog signaling in extraembryonic endoderm and ectoderm differentiation in ES embryoid bodies. AB - We previously demonstrated that a member of the Hedgehog gene family, Indian hedgehog (Ihh), is expressed in the visceral endoderm of EC and ES cell embryoid bodies and mouse embryos. Overexpression studies suggested that Ihh was involved in visceral endoderm differentiation. We now provide evidence for a Hh response in the embryoid body core and in the mesothelial layer of the visceral yolk sac. We also demonstrate that treatment of ES embryoid bodies with the Hh antagonists cAMP and forskolin results in downregulation of the Hh response and altered embryoid body differentiation. The outer endoderm layer undergoes a transition to parietal endoderm while formation of an embryonic ectoderm layer surrounding a cavity is inhibited. These treatments also result in a decrease in the expression of markers for the mesoderm derivatives, blood and endothelial cells. We present a model to explain how Ihh and BMP signaling may regulate extraembryonic endoderm and embryonic ectoderm differentiation. PMID- 10842065 TI - The Xenopus eomesodermin promoter and its concentration-dependent response to activin. AB - Eomesodermin is an essential early gene in Xenopus mesoderm formation and shows a morphogen-like response to activin. Here we define the regions of the Eomesodermin promoter required for mesodermal expression and for concentration dependent response to activin. We find an activin response element (ARE) located between -5.6 and -5.0 kb which contains two critical FAST2 binding sites. The ARE alone is necessary and sufficient for concentration-dependent response to activin. A 5.6 kb promoter recapitulates Eomes expression in normal mesoderm cells. A repressor element extinguishes Eomes expression in the endoderm. We relate our results to mesoderm patterning in early Xenopus development and to a mechanism of morphogen gradient response. PMID- 10842066 TI - Dorsal patterning defects in the hindbrain, roof plate and skeleton in the dreher (dr(J)) mouse mutant. AB - dreher is a spontaneous mouse mutation in which adult animals display a complex phenotype associated with hearing loss, neurological, pigmentation and skeletal abnormalities. During early embryogenesis, the neural tube of dreher mutants is abnormally shaped in the region of the rhomboencephalon, due to problems in the formation of a proper roof plate over the otic hindbrain. We have studied the expression of Hox/lacZ transgenic mouse strains in the dreher background and shown that primary segmentation of the neural tube is not altered in these mutants, although correct morphogenesis is affected resulting in misshapen rhombomeres. Neural crest derivatives from rhombomere 6, such as the glossopharyngeal ganglion, are defective, and the dorsal neural tube marker Wnt1 is absent from this segment. Selected trunk neural crest populations are also altered, as there is a lack of pigmentation in the thoracic region of mutant mice. Skeletal defects include abnormal cranial bones of neural crest origin, and improper fusion of the dorsal aspects of cervical and thoracic vertebrae. Taken together, the gene affected in the dreher mutant is responsible for correct patterning of the dorsal-most cell types of the neural tube, that is, the neural crest and the roof plate, in the hindbrain region. Axial skeletal defects could reflect inductive influence of the dorsal neural tube on proper fusion of the neural arches. It is possible that a common precursor population for both neural crest and roof plate is the cellular target of the dreher mutation. PMID- 10842067 TI - Characterization and expression analysis of an ancestor-type Pax gene in the hydrozoan jellyfish Podocoryne carnea. AB - We characterized a Pax gene from the hydrozoan Podocoryne carnea. It is most similar to cnidarian Pax-B genes and encodes a paired domain, a homeodomain and an octapeptide. Expression analysis demonstrates the presence of Pax-B transcripts in eggs, the ectoderm of the planula larva and in a few scattered cells in the apical polyp ectoderm. In developing and mature medusae, Pax-B is localized in particular endodermal cells, oriented toward the outside. Pax-B is not expressed in muscle cells. However, if isolated striated muscle tissue is activated for transdifferentiation, the gene is expressed within 1 h, before new cell types, such as smooth muscle and nerve cells, have formed. The expression data indicate that Pax-B is involved in nerve cell differentiation. PMID- 10842068 TI - Androcam, a Drosophila calmodulin-related protein, is expressed specifically in the testis and decorates loop kl-3 of the Y chromosome. AB - The Drosophila genome encodes a protein that is 68% identical to Drosophila calmodulin (Cam). We show here that this Cam-related gene is specifically expressed in the germ-line of the testis, leading to the name Androcam (Acam). Early in spermatogenesis Acam accumulates on one of the chromatin loops of the Y chromosome, kl-3. This association with kl-3 may indicate an RNA processing related role for Acam and/or could reflect an unusual storage/assembly function hypothesized for the Y loops. After meiosis Acam is detectable in developing sperm tail cytoplasm, where at least some of the protein is not tightly associated with tubulin. Late in spermiogenesis, some Acam staining overlaps the periphery of the investment cones, actin-containing structures hypothesized to support the motor function for cytoplasmic stripping of the tail. Acam cannot be detected in mature sperm by immunolocalization, but immunoblotting established that Acam is present in sperm stored in mated females, suggesting epitope masking during final maturation. Proteins more related to Acam than Cam are present in the testes of other Drosophila species and a mammalian species, the mouse. PMID- 10842069 TI - Expression of Meis and Pbx genes and their protein products in the developing telencephalon: implications for regional differentiation. AB - The Meis and Pbx genes encode for homeodomain proteins of the TALE class and have been shown to act as co-factors for other homeodomain transcription factors (Mann and Affolter, 1998. Curr. Opin. Genet. Dev. 8, 423-429). We have studied the expression of these genes in the mouse telencephalon and found that Meis1 and Meis2 display region-specific patterns of expression from embryonic day (E)10.5 until birth, defining distinct subterritories in the developing telencephalon. The expression of the Meis genes and their proteins is highest in the subventricular zone (SVZ) and mantle regions of the ventral telencephalon. Compared to the Meis genes, Pbx genes show a broader expression within the telencephalon. However, as is the case in Drosophila (Rieckhof et al., 1997. Cell 91, 171-183; Kurrant et al., 1998. Development 125, 1037-1048; Pai et al., 1998. Genes Dev. 12, 435-446), nuclear localized PBX proteins were found to correlate highly with Meis expression. In addition, DLX proteins co-localize with nuclear PBX in distinct regions of the ventral telencephalon. PMID- 10842070 TI - XSIP1, a Xenopus zinc finger/homeodomain encoding gene highly expressed during early neural development. AB - We have isolated a Xenopus homologue of the zinc finger/homeodomain-containing transcriptional repressor Smad-interacting protein-1 (SIP1) from mouse. XSIP1 is activated at the early gastrula stage and transcription occurs throughout embryogenesis. At the beginning of gastrulation, XSIP1 is strongly expressed in prospective neurectoderm. At the neurula stage, XSIP1 is highly expressed within the neural plate but weakly in the dorsal midline. At later stages of development transcripts are detected primarily within the neural tube and neural crest. In the adult, XSIP1 expression is detected at variable levels in several organs. PMID- 10842071 TI - Developmental regulation of the gradient of cftr expression in the rabbit heart. AB - Gradients of ion channels across the left ventricular free wall of the heart have been found for a number of repolarizing ion channels. Amongst these are the cAMP activated chloride channels encoded by cftr. In this report, we show that the epicardial (higher) to endocardial (lower) gradient of cftr mRNA found in adult rabbit hearts is not present in embryonic hearts. The gradient starts to develop shortly after birth, and over a period of 5-6 weeks increases to the levels found in the adult. This is the first report of the developmental regulation of any cardiac ion channel mRNA gradient. PMID- 10842073 TI - Xenopus FK 506-binding protein, a novel immunophilin expressed during early development. AB - FK 506-binding proteins (FKBPs) are a family of cytosolic proteins identified by virtue of their ability to bind the immunosuppressants FK 506 and rapamycin. While their function has been extensively studied in the immune system, little is known about their role during early embryonic development. Here we describe the cloning and expression of a new Xenopus FKBP (xFKBP). xFKBP encodes a 63-kDa protein that shares high sequence homology with mouse FKBP65. It is expressed maternally and becomes restricted after the gastrula stage to dorsal mesoderm and notochord. At the tailbud stage expression persists in the notochord and begins to accumulate in epidermis, branchial arches and developing somites. In adults, xFKBP mRNA is confined to the testis. PMID- 10842072 TI - Notch gene expression during pancreatic organogenesis. AB - Notch receptors are involved in regulating the balance between cell differentiation and stem cell proliferation during the development of numerous tissues (Artavanis-Tsakonas, S., Matsuno, K., Fortini, M. E., 1995. Notch signaling. Science 268, 225-232). Here the expression of all four vertebrate Notch genes, their ligands, and some down-stream targets is analyzed during mouse pancreatic organogenesis. Notch 1 is the first Notch gene expressed in the pancreatic epithelium, and coexpression with HES 1 suggests that the Notch 1 pathway is activated. Notch 2 expression follows later when pancreatic buds branch and is restricted to embryonic ducts, believed to be the source for endocrine and exocrine stem cells. Notch 3 and Notch 4 are expressed in pancreatic mesenchyme and later in endothelial cells. Together these descriptive data comprise a framework for understanding the cellular basis for Notch function during pancreatic development. PMID- 10842074 TI - Tissue-specific developmental expression of OAX, a Xenopus repetitive element. AB - Approximately 1% of the Xenopus laevis genome consists of highly repetitive DNA known alternatively as OAX (for Oocyte Activation in Xenopus), Satellite I, or Repetitive HindIII Monomer 2. Present as tandemly repeated units of approximately 750 base pairs, OAX encodes a family of small RNA species transcribed by RNA polymerase III. Although the subject of many of the classic studies on early embryonic gene regulation, reports on OAX expression remain contradictory and incomplete. Using whole-mount in situ hybridization and RNase protection assays, we have therefore examined in detail the expression pattern of OAX in Xenopus embryos of various stages. OAX is initially expressed during gastrula stages; by tailbud stages embryos display discrete zones of expression at the dorsal boundary of the cement gland, in the developing somites and differentiating skeletal muscle, as well as in the dorsal aspect of the neural tube. These data demonstrate that OAX is expressed in a dynamic pattern under tight spatial and temporal regulation. PMID- 10842075 TI - Expression of the vertebrate Slit gene family and their putative receptors, the Robo genes, in the developing murine kidney. AB - The slit (sli) gene, encoding a secreted glycoprotein, has been demonstrated to play a vital role in axonal guidance in Drosophila melanogaster by acting as a signalling ligand for the robo receptor (Rothberg, J.M., Jacobs, J.R., Goodman, C.S., Artavanis-Tsakonas, S., 1990. slit: an extracellular protein necessary for development of midline glia and commissural axon pathways contains both EGF and LRR domains. Genes Dev. 4, 2169-2187; Kidd, T., Bland, K.S., Goodman, C. S., 1999. Slit is the midline repellent for the robo receptor in Drosophila. Cell 96, 785-794). Multiple homologs of both sli and robo have been identified in vertebrates and are thought to play similar roles to their fly counterparts in neural development (Brose, K., Bland, K.S., Wang, K.H., Arnott, D., Henzel, W., Goodman, C.S., Tessier-Lavigne, M., Kidd, T., 1999. Slit proteins bind Robo receptors and have an evolutionarily conserved role in repulsive axon guidance. Cell 96, 795-806). Slit2 has been shown to bind Robo1, mediating both neuronal and axonal guidance in the developing central nervous system (CNS), (Brose et al., 1999; Hu, H., 1999. Chemorepulsion of neuronal migration by Slit2 in the developing mammalian forebrain. Neuron 23, 703-711). Importantly, both gene families display distinct expression patterns outside the CNS (Holmes, G.P., Negus, K., Burridge, L., Raman, S., Algar, E., Yamada, T., Little, M.H., 1998. Distinct but overlapping expression patterns of two vertebrate slit homologs implies functional roles in CNS development and organogenesis. Mech. Dev. 79, 57 72; Yuan, W., Zhou, L., Chen, J.H., Wu, J.Y., Rao, Y., Ornitz, D.M., 1999. The mouse SLIT family: secreted ligands for ROBO expressed in patterns that suggest a role in morphogenesis and axon guidance. Dev. Biol. 212, 290-306). Using in situ hybridization on metanephric explant cultures and urogenital tract sections, the expression patterns of Slit1, 2, 3 and Robo1 and 2 were investigated during murine metanephric development. Slit1 was expressed in the metanephric mesenchyme (MM) surrounding the invading ureteric tree (UT). Slit2 was expressed at the tips of the UT and both Slit2 and Slit3 were expressed at the far proximal end of the comma shaped and S-shaped bodies. Expression of Robo1 was initially diffuse throughout the MM, then upregulated in the pretubular aggregates, and maintained at the distal end of the comma and S-shaped bodies. Robo2 was detected in the induced MM surrounding the arborizing UT tips and later in the proximal end of the S-shaped bodies. Coincident expression of Robo1 with Slit1 in the metanephric mesenchyme and Robo2, Slit2 and Slit3 in the far proximal end of the S-shaped bodies was observed during metanephric development. PMID- 10842076 TI - Expression of Xenopus homologs of the beta-catenin binding protein pontin52. AB - Identification of pontin52 as an interaction partner of the Wnt/Wg signal transducer beta-catenin implicated a role for this protein in Wnt signaling. Here we describe the isolation of two Xenopus homologs of pontin52, Xpontin and Xreptin, and report the first expression pattern of vertebrate pontin52 homologs. Whole-mount in situ hybridization studies reveal a strong expression of Xpontin in neural crest cells and in later stages in different gastrointestinal organs. Xreptin is also expressed in neural crest cells, in particular in a subpopulation that give raise to the adrenal medulla. PMID- 10842077 TI - Tissue-specific expression of retinoic acid receptor isoform transcripts in the mouse embryo. AB - The three murine retinoic acid receptor (RAR) genes each contain two distinct promoters which give rise to protein isoforms differing in their N-terminal regions. This study used in situ hybridization to describe the expression patterns of RARalpha1, RARalpha2, RARbeta1/3, RARbeta2/4, RARgamma1 and RARgamma2 isoform transcripts during mouse embryogenesis. RARalpha1 transcripts are widely distributed, with the exception of the central nervous system. Highest expression is found in developing muscle, pituitary gland and various epithelia. On the other hand, RARalpha2 is essentially expressed along the spinal cord up to the hindbrain 7th rhombomere and in the 4th rhombomere, pons and developing basal ganglia (corpus striatum and pallidum). RARbeta2/4 transcripts account for most of the previously described RARbeta expression features being expressed specifically, or more prominently than RARbeta1/3, in foregut endoderm and its derivatives, olfactory and periocular mesenchyme, urogenital region, proximal limb bud mesenchyme and later within interdigital regions. RARbeta1/3 is more prominently expressed in the developing heart outflow tract mesenchyme, intervertebral disks, midgut loop mesenchyme and umbilical vessel walls. RARbeta1/3 and RARbeta2/4 are coexpressed in the developing corpus striatum. They exhibit, however, distinct dorsoventral distributions along the spinal cord and caudal hindbrain. RARgamma2 is the RARgamma isoform expressed at high levels in the caudal neural groove at embryonic day 8.5. At later stages, both RARgamma isoforms are essentially coexpressed, although the progressive restriction of RARgamma1 transcripts to craniofacial or limb precartilaginous condensations appears to precede that of RARgamma2. PMID- 10842078 TI - Alzheimer beta-amyloid precursor proteins display specific patterns of expression during embryogenesis. AB - The beta-amyloid precursor proteins (betaAPPs) are a family of glycosylated transmembrane proteins that include in their sequences the beta-amyloid peptide, a major component of the characteristic amyloid deposits or senile plaques found in the brains of Alzheimer's disease patients and aged Down's syndrome subjects. Various betaAPP isoforms, mainly betaAPP-695, betaAPP-714, betaAPP-751 and betaAPP-770, the number corresponding to the number of amino acids they encode, resulting from the alternative splicing of a single primary transcript have been described. Using oligonucleotides recognizing each of the four major Alzheimer's betaAPP mRNAs, we have found that each betaAPP mRNA displays a specific temporal and spatial pattern of expression. The prototype isoform betaAPP-695 occurs early in cells actively implicated in morphogenetic events, as those mesodermal cells invaginating at the level of the primitive streak, and it is later restricted to the neurectodermal (neural tube, neural crest and neurogenic placode) derivatives. By contrast, the longest isoform betaAPP-770 appears later and restricted to mesodermal and endodermal derivatives. The isoforms betaAPP-714 and betaAPP-751 are still expressed later than the other two isoforms and distributed ubiquitously, though betaAPP-714 transcripts predominate typically within the neural tube. PMID- 10842079 TI - The NK-2 homeobox gene scarecrow (scro) is expressed in pharynx, ventral nerve cord and brain of Drosophila embryos. AB - Members of the NK homeobox family have been widely conserved during evolution. Here we describe the sequence and expression of a novel Drosophila NK-2 homeobox gene, named scarecrow (scro), which shows considerable homology to vertebrate Nkx 2.1. During embryogenesis, scro expression is initially observed in the pharyngeal primordia and later maintained in the pharynx. During band germ retraction, scro expression appears in two bilateral clusters of procephalic neuroblasts that give rise to distinct neuronal clusters in the brain. In addition, scro expression is observed in segmental clusters of neuronal precursors in the ventral nerve cord. In larval stages, scro expression occurs in portions of the optic lobe regions. These observations indicate that scro and vertebrate Nkx2.1 share similarities both in terms of their sequence and their expression patterns. PMID- 10842080 TI - Xenopus frizzled 4 is a maternal mRNA and its zygotic expression is localized to the neuroectoderm and trunk lateral plate mesoderm. AB - We describe the identification and expression pattern of Xenopus frizzled 4 (Xfz4) gene during early development. Xfz4 protein presents characteristic features of a frizzled family member. The mature protein sequence of Xfz4 is 93% identical to murine Mfz4. Xfz4 is a maternal mRNA, its expression level remains constant during early development. The mRNA is first localized during gastrulation to the dorsal presumptive neuroectoderm. At the end of gastrulation, Xfz4 mRNA is detected in the dorso-anterior neuroectoderm. During neurulation, Xfz4 mRNA is expressed as a band on both side of the forebrain, and in the trunk lateral plate mesoderm. As development proceeds, expression of Xfz4 mRNA in the trunk lateral plate mesoderm decreases but persists in the forebrain. It is also expressed in the posterior unsegmented somitic mesoderm from late tail-bud stage onward. PMID- 10842081 TI - Analysis of Nedd4 expression during skeletal development in the mouse limb. AB - Nedd4, a ubiquitin-protein ligase, was originally identified as being down regulated during development of the mouse brain (Nedd denotes neural precursor cell expressed developmentally down-regulated) (Kumar, S., Tomooka, Y., Noda, M., 1992. Identification of a set of genes with developmentally down-regulated expression in the mouse brain. Biochem. Biophys. Res. Commun. 185, 1155-1161). Subtractive hybridization was used in an attempt to identify genes that are preferentially expressed early in skeletogenesis. Using this technique Nedd4 was identified multiple times. Northern blot analysis confirmed that Nedd4 is down regulated in the forelimb and hind limb. In situ hybridization was carried out to identify regions of the limb bud expressing Nedd4. Nedd4 is expressed weakly in condensing mesenchyme, and abundantly in proliferating and prehypertrophic chondrocytes, but is undetectable in hypertrophic chondrocytes. Primary cultures, which closely mimic in vivo chondrogenesis, were also used to demonstrate the stage-specific expression of Nedd4 during early skeletal development. PMID- 10842082 TI - Expression of Xenopus Daz-like protein during gametogenesis and embryogenesis. AB - Using a monoclonal antibody raised against Xenopus Daz-like protein (Xdazl), we showed that Xdazl is present in all stages of male and female germ cells except mature spermatozoa. Xdazl is not localized to any specific regions in early-stage embryos, in contrast to the strict localization of its mRNA in the germ plasm. Xdazl disappears after gastrulation but reappears in the primordial germ cells situated at the genital ridge. This is the first detailed report on the protein expression of a Daz-like gene during gametogenesis and embryogenesis in Xenopus, showing the difference in expression patterns of its mRNA and protein. PMID- 10842083 TI - SOX8 expression during chick embryogenesis. AB - We have isolated the SOX8 gene from the chicken embryo. This gene shows a high degree of sequence homology to SOX9 and SOX10. Detailed analysis of SOX8 expression by whole-mount in situ shows a dynamic and restricted expression pattern during chick development. SOX8 is expressed in the somitic derivative, the dermomyotome, the developing heart, pancreas, enteric neurone system, limb and the neural tube. This is the first detailed expression analysis of SOX8 in any species PMID- 10842084 TI - Expression of Crim1 during murine ocular development. AB - Crim1 (cysteine-rich motor neuron 1), a novel gene encoding a putative transmembrane protein, has recently been isolated and characterized (Kolle, G., Georgas, K., Holmes, G.P., Little, M.H., Yamada, T., 2000. CRIM1, a novel gene encoding a cysteine-rich repeat protein, is developmentally regulated and implicated in vertebrate CNS development and organogenesis. Mech. Dev. 90, 181 193). Crim1 contains an IGF-binding protein motif and multiple cysteine-rich repeats, analogous to those of chordin and short gastrulation (sog) proteins that associate with TGFbeta superfamily members, namely Bone Morphogenic Protein (BMP). High levels of Crim1 have been detected in the brain, spinal chord and lens. As members of the IGF and TGFbeta growth factor families have been shown to influence the behaviour of lens cells (Chamberlain, C.G., McAvoy, J. W., 1997. Fibre differentiation and polarity in the mammalian lens: a key role for FGF. Prog. Ret. Eye Res. 16, 443-478; de Iongh R.U., Lovicu, F.J., Overbeek, P.A., Schneider, M.D., McAvoy J.W., 1999. TGF-beta signalling is essential for terminal differentiation of lens fibre cells. Invest. Ophthalmol. Vis. Sci. 40, S561), to further understand the role of Crim1 in the lens, its expression during ocular morphogenesis and growth is investigated. Using in situ hybridisation, the expression patterns of Crim1 are determined in murine eyes from embryonic day 9.5 through to postnatal day 21. Low levels of transcripts for Crim1 are first detected in the lens placode. By the lens pit stage, Crim1 is markedly upregulated with high levels persisting throughout embryonic and foetal development. Crim1 is expressed in both lens epithelial and fibre cells. As lens fibres mature in the nucleus, Crim1 is downregulated but strong expression is maintained in the lens epithelium and in the young fibre cells of the lens cortex. Crim1 is also detected in other developing ocular tissues including corneal and conjunctival epithelia, corneal endothelium, retinal pigmented epithelium, ciliary and iridial retinae and ganglion cells. During postnatal development Crim1 expression is restricted to the lens, with strongest expression in the epithelium and in the early differentiating secondary fibres. Thus, strong expression of Crim1 is a distinctive feature of the lens during morphogenesis and postnatal growth. PMID- 10842085 TI - Dynamic expression of d-CdGAPr, a novel Drosophila melanogaster gene encoding a GTPase activating protein. AB - Small GTPases of the rho family function as signal transducer for extra-cellular stimuli to control cytoskeletal re-organization and a variety of other cellular processes including adhesion, proliferation and transcriptional regulation (Hall, A., 1998. RhoGTPases and the actin cytoskeleton. Science 279, 509-514). Usually widely expressed, their activities are tightly controlled by conformational changes induced by hydrolysis of the GTP bound molecule (Bourne H.R., Sanders D.A., 1990. The GTPase superfamily: a conserved switch for diverse cell functions. Nature 348, 125-132). Conversion of GTP to GDP relies on a rho intrinsic GTPase domain that requires GTPase activating proteins (GAPs) for potent activity (Lamarche, N., Hall. A., 1994. GAPs for rho-related GTPases. Trends Genet. 10, 436-440). Here we report on the identification of a novel Drosophila GAP gene, d-CdGAPr, encoding a protein related to mammalian CdGAPs. The gene is expressed throughout development as well as in adults. Spatio temporal transcription pattern of d-CdGAPr during embryogenesis is highly dynamic. Abundant in the pre-blastoderm embryo prior to the onset of zygotic transcription, messengers accumulate at the blastoderm posterior pole after cellularisation. During gastrulation and subsequent development, all cells accumulate low levels of d-CdGAPr RNA, while a few territories transiently display stronger expression. Sites of preferential expression include the posterior pole of the early cellular blastoderm, the neuro-ectoderm prior to neuroblast delamination, rows of epidermal cells in the most posterior part of thoracic and first abdominal segments and a ring of epidermal cells at the posterior end of the embryo. PMID- 10842086 TI - Cloning and developmental expression of mouse aldehyde reductase (AKR1A4). AB - The aldo-keto reductase superfamily catalyzes the reduction of a broad range of aldehydes and ketones to their corresponding alcohols. Here we report the cloning of the mouse aldehyde reductase cDNA and its embryonic pattern of expression. From stages E7.5 to E13.5 the gene encoding for this enzyme is expressed at high levels in several tissues, including the neural ectoderm, gut endoderm, somites, branchial arches, otic vesicles, limb buds, and tail bud. In adult mice aldehyde reductase was expressed in all tissues examined. PMID- 10842087 TI - Regulated expression of the proteoglycan SPOCK in the neuromuscular system. AB - SPOCK is prevalent in developing synaptic fields of the central nervous system (Charbonnier et al., 2000. Mech. Dev. 90, 317-321). The expression of SPOCK during neuromuscular junction (NMJ) formation was compared to agrin and acetylcholine receptor (AChR) distribution. SPOCK is detected within the myogenic masses during the early steps of embryonic development, and distributed in the cytoplasm of myotubes before coclustering with AChRs. In the adult, SPOCK is present in axons and is highly expressed by Schwann cells. SPOCK altered expression pattern after nerve lesioning, or cholinergic transmission blockade, strongly indicate that its cellular distribution at the NMJ depends on innervation. PMID- 10842088 TI - Expression of the RNA recognition motif-containing protein SEB-4 during Xenopus embryonic development. AB - RNA binding proteins play key roles in the post-transcriptional regulation of gene expression. Here we present the molecular cloning and spatio-temporal expression of Xseb-4, which codes for a putative RNA binding protein containing a single RNA recognition motif (RRM). XSEB-4 shares 60-65% identity with the mammalian SEB-4 proteins. Xseb-4 is strongly expressed maternally. Zygotic transcription is initiated in the early gastrula embryo in paraxial mesoderm that is fated to give rise to somites. During the course of gastrulation and neurulation Xseb-4 expression in somitic paraxial mesoderm is centered within the XmyoD expression domain. As development proceeds Xseb-4 expression is in addition initiated in the cardiac primordium and the lens vesicle. In the heart expression is confined to the myocardium. Thus, the RRM-containing putative RNA binding protein XSEB-4 is differentially expressed during embryonic development in Xenopus. PMID- 10842089 TI - Nestin expression during mouse eye and lens development. AB - Nestin is an intermediate filament protein, which is expressed predominantly in the developing central nervous system and skeletal muscles. In situ hybridization revealed that mouse nestin mRNA is in the optic stalk at 9.0 days post coitus (dpc) and in the lens vesicle at 10.0 dpc. From 14.5 dpc onward, nestin transcripts appear in lens fibers and neuroretina. Immunohistochemistry showed that nestin protein appears in the optic stalk at 9.5 dpc and in the posterior lens epithelium at 10.5 dpc. By 12.5 dpc, it is found in the lens, neuroretina, and optic stalk as well as in developing extrinsic ocular muscle, and it localizes in lens epithelium, optic disc, and optic nerve from 14.5 dpc to postnatal day 1. In adult eye, nestin protein appears in the optic nerve. PMID- 10842090 TI - Minireviews on bone: introductory remarks PMID- 10842091 TI - Bone formation and factors affecting this process. PMID- 10842092 TI - Molecular events caused by mechanical stress in bone. AB - The shape of bone changes as a result of bone remodeling corresponding to physical circumstances such as mechanical stress. The tissue which receives the loaded mechanical stress most efficiently is bone matrix. Recent studies revealed the function of osteocytes as mechanosensors in the early stage of bone remodeling. Loaded mechanical stress is converted to a series of biochemical reactions, and finally activates osteoclasts and osteoblasts to cause bone resorption and formation. Biochemical and molecular biological studies have recently resulted in the identification of the gene of which expression level is changed by mechanical stress. Nitric oxide (NO) and cAMP is secreted in response to mechanical stress in the immediate early stage. Genes encoding enzymes such as glutamate/aspartate transporter (GLAST), nitric oxide synthetase (NOS) and prostaglandin G/H synthetase (PGHS-2) are identified as mechanical stress responsive. The expression level of IGF-I is enhanced under the control of PTH/PTHrP. The expression of c-fos is increased by loading of mechanical stress. AP1, a heterodimer of c-FOS/c-JUN, functions as a transcription factor of downstream gene(s). Elements including AP1 sites, cyclic AMP response elements (CRE) and shear stress response elements (SSRE) are found in the promoter region of mechanical stress-response genes. The enhanced expression of osteopontin (OPN) in the osteocytes of bone resorption sites was demonstrated by in situ hybridization and immunohistochemistry and transdifferentiation of chondrocytes with the abundant expression of BMP-2 and -4 in the process of distraction osteogenesis was observed. PMID- 10842093 TI - Integrins and signaling in osteoclast function. AB - Integrins are heterodimeric adhesion receptors that mediate cell-matrix and cell cell interactions. Osteoclasts highly express the alphavbeta3 integrin, which binds to a variety of extracellular matrix proteins including vitronectin, osteopontin and bone sialoprotein. RGD-containing peptides, RGD-mimetics and alphavbeta3 blocking antibodies inhibit bone resorption in vitro and in vivo, suggesting that this integrin plays an important role in osteoclast function. RGD containing peptides were shown to raise cytosolic calcium in osteoclasts. Furthermore, several signaling and adaptor molecules were found to be involved in alphavbeta3 integrin-dependent signaling pathways, including phosphatidylinositol 3-kinase, c-Src, PYK2 and p130(cas). In addition, cytoskeletal molecules such as paxillin, vinculin, gelsolin and F-actin are recruited to adhesion contacts upon integrin activation. Many of these molecules signaling and cytoskeletal localize to the sealing zone of actively resorbing osteoclasts, suggesting that they play a role in linking the adhesion of osteoclasts to the bone matrix with the cytoskeletal organization and the polarization and activation of these cells for bone resorption. PMID- 10842094 TI - Osteoclast formation and resorption. PMID- 10842095 TI - Exon skipping mutation in the COL9A2 gene in a family with multiple epiphyseal dysplasia. AB - Previous linkage analysis (Briggs, M.D., Choi, H.-C., Warman, M.L. et al., 1994. Genetic mapping of a locus for multiple epiphyseal dysplasia (EDM 2) to a region of chromosome 1 containing a type IX collagen gene. Am. J. Hum. Genet. 55, 678 684) in a large English family with multiple epiphyseal dysplasia established the EDM2 locus, a region of chromosome 1 containing the COL9A2 collagen gene. We now report that affected members of this family are heterozygous for a single base transversion (T-->G) at the sixth position of the intron 3 splice donor of COL9A2. The mutation leads to skipping of exon 3 during splicing, and results in a 36-nucleotide deletion in COL9A2 transcripts derived from the mutant allele. Skipping of exon 3 predicts an in-frame deletion of 12 amino acid residues within the COL3 domain of the alpha2(IX) chain. This is the fifth instance of an exon 3 deletion within the COL3 region of collagen IX heterotrimers causing the MED phenotype, as yet the only type IX collagen defect identified in this disorder. Electron microscopy (EM) of chondrocytes obtained from articular cartilage of one affected individual in the family demonstrated normal appearing rough endoplasmic reticulum (RER). In addition, the articular cartilage matrix did not show any gross abnormalities in the quantity or caliber of collagen fibrils. PMID- 10842096 TI - Catabolism of newly synthesized decorin by explant cultures of bovine ligament. AB - The catabolism of newly synthesized decorin by explant cultures of bovine collateral ligament was investigated. The tissue was placed in explant culture for 6 days then incubated with radiolabeled sulfate for 6 h and replaced in culture for 5 days to allow for the loss of the radiolabeled large proteoglycan. The metabolic fate of the remaining radiolabeled decorin present in the matrix of the tissue over the next 9-day period was determined. It was shown that this pool of decorin was lost from ligament explant cultures either directly into the culture medium or taken up and degraded within the cells of the tissue. The intracellular degradation of the radiolabeled pool of decorin by ligament explant cultures was shown to result in the generation of [35S]sulfate. This process required metabolically active cells and involved the lysosomal system since sulfate generation was inhibited when cultures were maintained at 4 degrees C or in the presence of either 10 mM ammonium chloride or 0. 05 mM chloroquine. The inhibition of intracellular processing of decorin resulted in an increase in the rate of loss of this proteoglycan into the medium of the cultures. The inhibition of intracellular degradation of decorin was reversible on incubation of the explant cultures at 37 degrees C or removal of ammonium chloride from the culture medium. After removal of the ammonium chloride from the culture medium the rate of intracellular catabolism was greater than that observed in cultures maintained in medium alone, which suggested that there was an intracellular accumulation of native and/or partially degraded material within the cells. PMID- 10842097 TI - Analysis of the ex vivo specificity of human gelatinases A and B towards skin collagen and elastic fibers by computerized morphometry. AB - Cutaneous aging and chronic exposure to UV irradiation leads to alterations in the appearance and biochemical composition of the skin. Members of the MMP family have been involved in the destruction of the extracellular matrix. Among them, gelatinases A and B were found to display elastolytic activity, in vitro. In this study, we first determined the ex vivo elastolytic potential of both endopeptidases, using human skin tissue sections and computerized morphometric analyses, and compared it with those of neutrophil elastase. In such conditions, gelatinase B (50 nM) induced 50% elastolysis. The percentage of elastic fibers degraded by gelatinase A (10-100 nM) never exceeded 10%. Elastolysis by gelatinase B and leukocyte elastase was characterized by a decrease in fiber length and an increase in the average diameter of the fibers. In addition, gelatinase B exhibited fibrillin-degrading activities. On the contrary, gelatinase A (50 nM) elicited up to 50% hydrolysis of collagen fibers, preferentially degrading type III collagen fibers. Gelatinase B did not promote any collagen degrading activity. Our data suggested that in vivo gelatinases could disrupt most extracellular matrix structures of human skin. Gelatinase B and to a much lesser extent, gelatinase A would degrade components of the elastic fibers network while gelatinase A, but not gelatinase B, would alter mostly collagen fibers and also degrade constituents of the dermo-epidermal junction. PMID- 10842098 TI - Developmental expression of dermatan sulfate proteoglycans in the elastic bovine nuchal ligament. AB - The nuchal ligament of bovines is a useful system in which to study elastic fibre formation since it contains up to 83% elastin and undergoes a period of rapid elastinogenesis during the last trimester of fetal development and in the first four post-natal months. To identify proteoglycans (PGs) which may be involved in this process we initially investigated changes in the glycosaminoglycan (GAG) profiles during nuchal ligament development. In contrast to the collagenous Achilles tendon, nuchal ligament exhibited: (a) elevated hyaluronan (HA) levels in the peak period of elastin-associated microfibril (fibrillin) synthesis (130 200 days) which precedes elastinogenesis; and (b) markedly increased synthesis of a glucuronate-rich copolymeric form of dermatan sulfate (DS) in the period corresponding to elastin formation (200-270 days). Analysis of DSPGs isolated from 230-day nuchal ligament showed that this copolymer was predominantly associated with a glycoform of biglycan which was specifically elevated at this stage in development. This finding was consistent with Northern blot analysis which showed that steady-state biglycan mRNA levels increased significantly during the elastinogenic period. In contrast, the mRNA levels for decorin, the only other DSPG detected in this tissue, declined rapidly after 140 days of fetal development. In conclusion, the results suggest that HA may play a role in microfibril assembly and that a specific glycoform of biglycan may be associated with the elastinogenic phase of elastic fibre formation. PMID- 10842099 TI - Chain specificity assignment of monoclonal antibodies to human laminins by using recombinant laminin beta1 and gamma1 chains. AB - In the present study, the chain specificity of 16 commonly used monoclonal antibodies to human laminin(s) was analysed by using recombinant laminin beta1 and gamma1 chains. By ELISA, all antibodies reacted with purified placenta laminin, and most antibodies recognised either recombinant beta1 or gamma1 chains. Reactivity and chain specificity was confirmed against the recombinant chains in Western blotting under non-reducing conditions, and only a few antibodies were reactive under reducing conditions. Most antibodies were able to immunoprecipitate associated laminin beta1/gamma1 chains from platelet lysates. Based on these results and data from the literature, a tentative epitope map is presented. PMID- 10842100 TI - Localization of pN-type IIA procollagen on adult bovine vitreous collagen fibrils. AB - Type II procollagen is synthesized in long (type IIA) and short (type IIB) forms because of alternative splicing of mRNA; the long form containing an additional cysteine-rich domain in the amino-propeptide. An antiserum (IIA) that recognizes this domain was used for immunolocalization studies on adult bovine vitreous at light and electron microscopic levels and for Western blot analyses. The immunolocalization studies revealed labelling by the IIA antiserum of the vitreous collagen fibrils. This labelling was removed by prior extraction of the fibrils with 6 M guanidine hydrochloride (GuHCl) and the extract was shown to contain pN-type IIA procollagen. Adult vitreous collagen fibrils are coated with pN-type IIA procollagen, a finding with potential implications for vitreous collagen fibril structure and function. PMID- 10842101 TI - Rapid genotype analysis of the matrix metalloproteinase-1 gene 1G/2G polymorphism that is associated with risk of cancer. AB - A bi-allelic polymorphism in the promoter of the human matrix metalloproteinase-1 gene has been reported. It has been found to have a functional effect on the promoter strength and to be associated with risk of cancers. The polymorphism is due to an insertion/deletion of a guanine, and conventional methodologies for genotyping this polymorphism are time-consuming and expensive. A rapid genotyping method based on restriction endonuclease digestion is reported here. PMID- 10842102 TI - The mouse lysyl oxidase-like 2 gene (mLOXL2) maps to chromosome 14 and is highly expressed in skin, lung and thymus. AB - The predicted amino acid sequence derived from a mouse expressed sequence tag (EST) contig contained two domains that are highly conserved among members of the lysyl oxidase gene family: a copper binding-site with four histidines and a catalytic domain that includes a tryptophan residue. This new cDNA sequence showed the highest level of sequence homology with the human loxl2 cDNA and suggested that it encoded the mouse equivalent of hLOXL2. The mLOXL2 gene was mapped to chromosome 14 by radiation hybrid analysis. The mLOXL2 locus was tightly linked with a LOD score over 9 to the marker D14Mit32. The mLOXL2 gene is expressed as a 4-kb mRNA in almost all tissues analyzed, with highest levels of mRNA in skin, lung and thymus. PMID- 10842104 TI - Online AJO/DO becomes a member benefit PMID- 10842103 TI - Chromosomal mapping of Adam9, Adam15 and Adam21. AB - Adam9, Adam15 and Adam21, genes encoding members of the ADAM or MDC family of metalloproteases, have been mapped to mouse chromosomes 8, 1, and 12, respectively, using an interspecific cross. The mapping of these mouse loci and the extrapolated loci for their human orthologs may facilitate the mapping of diseases involving these genes. PMID- 10842105 TI - Evaluation of profile esthetic change with mandibular advancement surgery. AB - Our purpose was to investigate the impact of mandibular advancement surgery on profile esthetics and to attempt to define guidelines that could be of value to the clinician in predicting profile esthetic change. The sample consisted of 34 patients who had been treated with a combination of orthodontics and mandibular advancement surgery without genioplasty. Initial (pretreatment) and final (posttreatment) cephalometric radiographs of each patient were used to produce silhouette images and to quantify skeletal changes that occurred with surgery. The images were displayed randomly to lay persons and orthodontic residents who were asked to score the esthetics of each profile. On average, after mandibular advancement surgery, B point moved forward 5.0 mm (SD = 2.6 mm) and downward 4.7 mm (SD = 3.1 mm), and the ANB angle decreased 3.0 degrees (SD = 1.6 degrees ) Graphical analysis and results of paired t tests revealed that for patients with an initial ANB angle >/= 6 degrees, a consistent improvement in profile esthetics was seen following surgery (P 52%) for all experimental groups. Tooth preparations with greater convergence and internally relieved castings recorded a better marginal fit. CONCLUSION: The casting internal adjustment technique with use of duplicated stone dies and a disclosing agent substantially reduced marginal fit discrepancy. PMID- 10842131 TI - Effects of surface treatments on shear bond strengths between a resin cement and an alumina core. AB - STATEMENT OF PROBLEM: Although bonding to all-ceramic restorations is desirable, there is little information on the use of resin cements containing a phosphate monomer, and the importance of different surface treatments on their adhesion to high-strength core materials. PURPOSE: This study attempted to determine the shear bond strength values between Panavia 21 resin cement (Kuraray) and an alumina core material (In-Ceram) after 3 surface treatments and the application of a silane coupling agent. MATERIAL AND METHODS: Forty-five In-Ceram cylindrical rods were fabricated and assigned to 3 groups. Group I specimens were treated with a 9.5% hydrofluoric (HF) acid, group II with a 5% HF acid, and group III were sandblasted. All specimens were coated with a silane coupling agent (Cavex Clearfil Photobond and Activator) before cementation with Panavia 21 to sandblasted nickel-chromium rods. As a control, group IV consisted of 8 porcelain (Vitadur Alpha) rods treated with a 5% HF acid and silane. All specimens were subjected to a load of 1.2 kg during cementation, then stored under water for 36 hours. A jig mounted on a Hounsfield Universal Testing machine was used at a crosshead speed of 0.5 mm/min to test the shear bond strengths. RESULTS: The results were 14.65 +/- 4.64 MPa for group I, 18.03 +/- 6.13 MPa for group II, and 22.35 +/- 5.98 MPa for group III In-Ceram specimens; and 18.05 +/- 8.46 MPa for control (group IV). CONCLUSION: The use of Panavia 21 resin cement and a silane coupling agent can achieve a successful bond between either sandblasted or 5% HF acid-etched In-Ceram core material. PMID- 10842132 TI - Impression procedure for creating a partial auricular prosthesis. AB - Severe anatomic undercuts associated with tissue remnants or a defect site can often be an obstacle in achieving an accurate impression for a facial prosthesis. This article describes a procedure that overcomes the problems encountered in achieving an accurate impression. The distinct properties of 2 impression materials are applied to create a 3-piece impression that can be reassembled to develop an accurate stone master cast. PMID- 10842133 TI - Replacement of an obturator section of an existing two-piece implant-retained edentulous obturator. AB - There is an increasing number of people in the community who have postablative surgery for tumors of the maxilla. Postsurgical defects these individuals have are usually restored by means of a complete or partial denture obturator with various materials, including resilient silicone extensions. These patients require long-term maintenance of their obturator prostheses, which must be considered in the context of their general health and ongoing medical care. When a resilient silicone bulb is used to obturate the defect, the silicone sometimes deteriorates, whereas the denture base remains functional. This article describes a simple procedure to construct a replacement, resilient silicone bulb obturator while retaining the original complete or partial denture base. PMID- 10842134 TI - Clear acrylic resin device for orientation and placement of a small facial prosthesis. AB - A small-sized facial prosthesis can be difficult to orient accurately in correct relationship to a defect. This problem may be reduced by embedding a magnet in the prosthesis and constructing a magnetic holding device from clear acrylic resin. This device can be used as a stand during adhesive application to reduce adhesive contamination and acts as orientation guide during prosthesis placement. PMID- 10842135 TI - Hardness of celluloid strip-finished or polished composite surfaces with time. AB - STATEMENT OF PROBLEM: An in-vitro study revealed that a celluloid strip-finished composite surface discolored more than the polished composite surface. Thus, the celluloid strip-finished composite surface may not cure enough compared with the polished composite surface. PURPOSE: This study tested the hypothesis that the celluloid strip-finished composite surface did not cure enough compared with the polished composite surface. METHODS AND MATERIAL: The composite was placed in a 1.5-mm thick aluminum mold and the upper surface was covered with a celluloid strip. Composite was light cured for 60 seconds, then a layer of approximately 200 microm thick was ground away from the lower surface and polished. The hardness of the upper composite surface that was polymerized under a celluloid strip and the polished lower surface were measured with Vickers hardness measuring instrument 15 minutes, 6 hours, and 6 days after light curing. The hardnesses of polished and celluloid strip-finished surfaces were compared using a paired t test. One-way ANOVA and Tukey was used for tests for the significant differences in hardnesses between 15 minutes, 6 hours, and 6 days after light curing for a given surface. RESULTS: Microhardness of the celluloid strip finished composite surface was 380.6 N/mm(2) at 15 minutes, 442.5 N/mm(2) at 6 hours, and 519.2 N/mm(2) at 6 days after light curing. Microhardnesses of the polished composite surface was 476.6 N/mm(2) at 15 minutes, 511.7 N/mm(2) at 6 hours, and 535.0 N/mm(2) at 6 days after light curing. The hardness of the celluloid strip-opposed composite surface was significantly lower than that of the polished surface 15 minutes and 6 hours after light curing (P <. 001). There was no difference in hardness between the celluloid strip-opposed surface and the polished surface on the sixth day. CONCLUSION: The hardness of the celluloid strip-opposed composite surface was lower than that of the polished surface at 15 minutes and at 6 hours after light curing. However, there was no difference in microhardness in 6 days. PMID- 10842136 TI - Effect of surface roughness of porcelain on adhesion of bacteria and their synthesizing glucans. AB - STATEMENT OF PROBLEM: In some instances of porcelain restoration, refinishing is inevitable. In terms of plaque accumulation on porcelain, refinishing could be a substitute method for glazing. PURPOSE: This study compared the amount of adhesion of plaque components (bacterial cells and glucans) on porcelain disks with various degrees of surface roughness to assess the effects of surface roughness on the amount of plaque accumulation. MATERIAL AND METHODS: Radiolabeled cell suspensions were incubated with porcelain disks for 3, 8, and 24 hours at 37 degrees C, and the amounts of adhered cells and glucans were measured by using a liquid scintillation method. RESULTS: The amount of cells and glucans adhered on porcelain increased with incubation time. The surface roughness value and the amount of plaque adhesion decreased with the increase in polishing level. However, the greatest amount of plaque was adhered on glazed surfaces, although their surfaces were smoother than the surfaces polished with 120- or 600-grit abrasive papers. CONCLUSION: With the exception of glazed surfaces, a positive correlation between surface roughness and the amount of plaque accumulation was observed. Repolishing with a diamond paste would not induce problems of plaque accumulation, compared with an intact glazed surface. PMID- 10842137 TI - In vitro dentinal surface reaction of 9.5% buffered hydrofluoric acid in repair of ceramic restorations: a scanning electron microscopic investigation. AB - STATEMENT OF PROBLEM: Fracture of porcelain is a relatively common clinical misfortune. Recent research has indicated that strong bonds can be formed between composite and dental porcelain. Porcelain surfaces are etched with hydrofluoric acid and treated with silane coupling agents before composite application. The question is how exposed dentin may react to etching with hydrofluoric acid. PURPOSE: This investigation examined the effect of 9.5% buffered hydrofluoric acid, of 36% o-phosphoric acid alone and in combination on the surface structure of cut human dentin. MATERIAL AND METHODS: Human molar teeth were sectioned in approximately 0.8-mm thick slices and treated with different acids or their combinations. Application periods were 10, 60, and 180 seconds. Specimens were processed for SEM and for energy-dispersive x-ray (EDX) microanalysis. RESULTS: The smear layer on the surface of sectioned dentin was not completely removed by hydrofluoric acid alone and that a dense amorphous precipitate was formed on the peritubular zone. Starlike structures in dentinal tubules were visible. EDX analysis revealed different fluoride content on the treated surface, dependent on the etchant used. CONCLUSION: Topical application of hydrofluoric acid appeared to provide a dentinal surface with an amorphous precipitate of fluoride. This layer may be important both for resistance of dental caries in dentin and for bonding reactions. PMID- 10842138 TI - Evaluation of dental erosion in patients with gastroesophageal reflux disease. AB - STATEMENT OF PROBLEM: The cause of dental erosion may be difficult to establish because of its many presentations. Determination of the cause is an important aspect of diagnosis before extensive prosthodontic rehabilitation. PURPOSE: This cross-sectional study evaluated the association between loss of tooth structure as a result of dental erosion and gastroesophageal reflux disease. MATERIAL AND METHODS: Twenty consecutive adult dentate subjects referred to the Division of Gastroenterology for investigation of gastroesophageal tract disease were also evaluated for signs of dental erosion. All subjects underwent a dental evaluation that included a patient history to determine potential etiologic factors responsible for dental erosion. Subjects were examined clinically to quantify loss of tooth structure using a Tooth Wear Index (TWI). Endoscopic examination and 24-hour pH manometry were carried out to determine which subjects met the criteria for gastroesophageal reflux disease (GERD). Scores for maxillary versus mandibular dentition and anterior versus posterior dentition were also compared. Data were analyzed with the Kruskal-Wallis test (P =.004). RESULTS: Ten subjects were diagnosed with GERD and 10 subjects had manometry scores below the level indicating GERD. Overall, subjects diagnosed with GERD had significantly higher TWI scores compared with control subjects (mean difference = 0.6554; P =.004). GERD subjects had higher TWI scores in all quadrants, except in the mandibular anterior region where there was no difference. CONCLUSION: The results indicated that a relationship exists between loss of tooth structure, as measured by the TWI index, and the occurrence of GERD in this group of subjects. PMID- 10842139 TI - Accurate procedure for simultaneous registration of gingival emergence profile and maximal intercuspal position for metal ceramic restorations. AB - This article describes a procedure that simultaneously registers maximal intercuspal position and gingival emergence profile. The materials used in this procedure are inexpensive, readily available, and easy to use. The clinician can inspect the framework, cast, and tissue profile before sending them to the laboratory. When this method is properly used, it allows better communication with the dental technician, saves chair time, and reduces the number of laboratory procedures as currently practiced. PMID- 10842140 TI - Simplified technique for refabrication of cast posts and cores. AB - This article describes a simple procedure of recording contours of an existing cast post and core that needs to be reconstructed, using a polyvinyl siloxane material as a matrix for fabrication of a new cast post and core in a relatively short time with reduced cost. The procedure involves the use of an acrylic resin material under pressure, which minimizes void formation in the post-and-core pattern. PMID- 10842141 TI - Simple procedure for mouth guard identification for school-aged children. PMID- 10842142 TI - Method to contour the intaglio surface of an immediate denture when using a chairside soft reline procedure after tooth extraction. PMID- 10842143 TI - Inversion of gloves to prevent contamination. PMID- 10842144 TI - Presidential address: "These united colonies are...". PMID- 10842145 TI - Epidural cooling for spinal cord protection during thoracoabdominal aneurysm repair: A five-year experience. AB - PURPOSE: We developed and applied a method for providing regional spinal cord hypothermia with epidural cooling (EC) during thoracoabdominal aneurysm (TAA) repair. Preliminary results indicated significant reduction in spinal cord ischemic complications (SCI), compared with historical controls, and a 5-year experience with EC was reviewed. METHODS: From July 1993 to September 1998, 170 patients with thoracic aneurysms (n = 14; 8.2%) or TAAs (types I and II, n = 83 [49%]; type III, n = 66 [39%]; type IV, n = 7 [4.1%]) were treated with EC. An earlier aneurysm resection was noted in 44% of patients, an emergent operation was noted in 20% of patients, and an aortic dissection was noted in 16% of patients. The EC was successful (mean cerebrospinal fluid [CSF] temperature at cross-clamp, 26.4 +/- 3 degrees C) in 97% of cases, with all 170 patients included in an intention-to-treat analysis. The operation was performed with a clamp/sew technique (98% patients) and selective (T(9) to L(1) region) reimplantation of intercostal vessels. Clinical and EC variables were examined for association with operative mortality and SCI by means of the Fischer exact test, and those variables with a P value less than.1 were included in multivariate logistic regression analysis. RESULTS: The operative mortality rate was 9.5% and was weakly associated (P =.07) with SCI; postoperative cardiac complications (odds ratio [OR], 35. 3; 95% CI, 5.3 to 233; P <.001) and renal failure (OR, 32.2; 95% CI, 6.6 to 157; P <.001) were the only independent predictors of postoperative death. SCI of any severity occurred in 7% of cases (type I/II, 10 of 83 [12%]; all other types, 2 of 87 [2.3%]), versus a predicted (Acher model) incidence of 18.5% for this cohort (P =. 003). Half the deficits were minor, with good functional recovery, and devastating paraplegia occurred in three patients (2.0%). Independent correlates of SCI included types I and II TAA (OR, 8.0; 95% CI, 1.4 to 46.3; P =.021), nonelective operation (OR, 8.3, 95% CI, 1.8 to 37.7; P =.006), oversewn T(9) to L(2) intercostal vessels (OR, 6.1; 95% CI, 1.3 to 28.8; P =.023), and postoperative renal failure (OR, 23.6; 95% CI, 4.4 to 126; P <.001). These same clinical variables of nonelective operations (OR, 7.7; 95% CI, 1.4 to 41.4; P =.017), oversewn T(9) to L(2) intercostal arteries (OR, 9.7; 95% CI, 1.5 to 61.2; P =.016), and postoperative renal failure (OR, 20.8; 95% CI, 3.0 to 142.1; P =.002) were independent predictors of SCI in the subgroup analysis of high-risk patients, ie, patients with type I/II TAA. CONCLUSION: EC has been effective in reducing immediate, devastating, total paraplegia after TAA repair. A strategy that combines the neuroprotective effect of regional cord hypothermia, avoiding the sacrifice of potential spinal cord blood supply, and postoperative adjuncts (eg, avoidance of hypotension, CSF drainage) appears necessary to minimize SCI after TAA repair. PMID- 10842146 TI - Gender and carotid endarterectomy: does it matter? AB - OBJECTIVE: Multiple large series have retrospectively identified female gender as a risk factor for perioperative stroke and death after carotid endarterectomy (CEA). METHODS: Data for all patients who underwent CEA at a single institution from January 1990 to December 1998 were entered into a computerized vascular registry and form the basis of this report. RESULTS: A total of 1298 CEA procedures were performed, of which 520 (40%) were in women and 778 (60%) in men. The mean age was 69.8 +/- 8.7 years for men and 71.2 +/- 8.5 years for women (P <.001). Cardiac risk factors significantly varied among the two groups, with women more likely to have diabetes (42% vs 36%) and hypertension (77% vs 66%), whereas tobacco history was higher among men (85% vs 71%) (P <.05 for all). Female patients were more likely to be asymptomatic at presentation (men, 44% vs women, 51%; P =.022). Postoperative myocardial infarction occurred in eight patients (0.6%) with no differences between men (0.4%) and women (1.0%) (P = not significant). For all adverse postoperative cardiac events (myocardial infarction, congestive heart failure, or arrhythmia), the incidence was 1.9% (25 patients), again with no differences between men (1.5%) and women (2. 5%) (P = not significant). There were 25 postoperative neurologic events (19 strokes, six transient ischemic attacks) among the entire cohort (1.9%), of which 16 were in men (2.1%) and nine in women (1. 6%; P = not significant). The overall postoperative stroke rate was 1.5% (13 [1.7%] of 778 men; 6 [1.2%;] of 520 women; P = not significant). Total operative mortality was 0.3% (3 [0.4%] of 778 men; 1 [0.2%] of 778 women; P = not significant). Late recurrent stenosis requiring operation developed in 14 patients (1.1%) during follow-up (6 [0.8%] of 778 men; 8 [1.5%] of 520 women; P =.19). CONCLUSIONS: Although there is significant variability in cardiac risk factors and presentation, female gender is not a risk factor for stroke, death, or cardiac morbidity after CEA. Women are not at higher risk for reoperation for recurrent stenosis. PMID- 10842147 TI - Management of ischemic heel ulceration and gangrene: An evaluation of factors associated with successful healing. AB - OBJECTIVE: The objective of this study was to determine the effectiveness of treatment of nonhealing heel ulcers and gangrene and to define those variables that are associated with success. METHODS: A multi-institutional review was undertaken at four university or university-affiliated hospitals of all patients with wounds of the heel and arterial insufficiency, which was defined as absent pedal pulses and a decreased ankle/brachial index (ABI). Risk factors, hemodynamic parameters, and arteriographic findings were statistically analyzed to determine their effect on wound healing. Life-table analysis was used to assess graft patency and wound healing. RESULTS: Ninety-one patients (57 men, 34 women) were treated for heel wounds that did not heal for 1 to 12 months (62% of nonhealing wounds, 3 months or longer). The mean preoperative ABI was 0.51, and 31% of wounds were infected. Of the patients, 55% had impaired renal function (Cr > 1.5), with 24% undergoing dialysis, 70% had diabetes, and 64% smoked cigarettes. Treatment was topical wound care for all patients and operative wound debridement in 50%. Infrainguinal bypass was performed for 81 patients, 4 had inflow procedures, 3 had superficial femoral artery percutaneous transluminal angioplasty, and 3 had primary below-knee amputation. Postoperatively, 85% of patients had in-line flow to the foot with at least a single patent vessel, 66% had a pedal pulse, and the mean ABI improved by 0.40, to 0.91. Follow-up ranged from 1 to 60 months (mean, 21 months), and 77 patients (85%) are currently alive. In 66 patients (73%), the wounds healed-all within 6 months (mean, 3 months). For 14 (16%) the wounds had not healed, and 11 patients (11%) underwent below-knee amputation. By life-table analysis, limb salvage was 86% at 3 years. During follow-up, 75 infrainguinal bypasses (91%) remained patent (3 secondarily) and 6 occluded, with primary assisted patency of 87% at 3 years. All wounds in patients with occluded grafts failed to heal. Variables found to be statistically significant in predicting healing included normal renal function (95% healed vs 55% nonhealed, P <.002), a palpable pedal pulse (85% healed vs 42%, P <.0015), a patent posterior tibial artery past the ankle (86% healed vs 57%, P <.02), and the number of patent tibial arteries after bypass to the ankle (P <.0001). Neither the ABI nor the presence of infection (defined as positive tissue cultures or the presence of osteomyelitis), diabetes, or other cardiovascular risk factors influenced the outcome. CONCLUSIONS: Complete wound healing of ischemic heel ulcers or gangrene may require up to 6 months, and short-term graft patency is of minimal benefit. Successful arterial reconstruction, especially a patent posterior tibial artery after bypass, is effective in treating most heel ulcers or gangrene. Patients with impaired renal function are at increased risk for failure of treatment, but their wounds may successfully heal and they should not be denied revascularization procedures. PMID- 10842148 TI - Arm vein conduit is superior to composite prosthetic-autogenous grafts in lower extremity revascularization. AB - PURPOSE: Various alternative conduits have been used for lower extremity revascularization when an adequate ipsilateral greater saphenous vein is absent. This study compared the effectiveness of all-autogenous multisegment arm vein bypass grafts with that of composite grafts composed of combined prosthetic and autogenous conduits. METHODS: One hundred fifty-three lower extremity revascularization procedures performed between 1990 and 1998 were followed up prospectively using a computerized vascular registry. The grafts were composed of spliced arm vein segments with venovenostomy in 122 and of composite prosthetic autogenous conduit in 31. Arm vein conduit was prepared by means of intraoperative angioscopy for valve lysis and identification of luminal abnormalities in 47.7% of cases. RESULTS: Bypass graft configurations were as follows: femoropopliteal (12 arm vein, 2 composite); femorotibial (75 arm vein, 23 composite); femoropedal (14 arm vein, 6 composite), and popliteo-tibial/pedal (21 arm vein, 0 composite). The indication for surgery was limb salvage in 98% and disabling claudication in 2% of cases. The mean follow-up was 25.1 months (range, 1 month to 7.9 years). Overall survival at 4 years was 51%. Overall patency and limb salvage rates were as follows: primary patency, at 1 year-arm vein, 76.9% +/- 4.8%; composite, 59. 5% +/- 9.6% (P =.02); at 3 years-arm vein, 70.0% +/- 8.0%; composite, 43.7% +/- 12.4% (P <.01); and at 5 years-arm vein, 53.8% +/- 8.7%; composite, 0%; secondary patency, at 1 year-arm vein, 77.5% +/- 4. 6%; composite, 59.8% +/- 9.5% (P =.02); at 3 years-arm vein, 70.7% +/- 7.5%, composite, 44.9% +/- 13.1% (P <.01); at 5 years-arm vein, 57.7% +/- 8.0%; composite, 0%; limb salvage, at 1 year-arm vein, 89. 3% +/- 3.7%; composite, 73.9% +/- 8.9% (P <.01); at 3 years-arm vein, 80.5% +/- 7.0%; composite, 49.6% +/ 14.3% (P <.01); at 5 years-arm vein, 76.3% +/- 9.9%; composite, 0%. CONCLUSION: In this study, multisegment autogenous arm vein was used successfully in a wide variety of lower extremity revascularization procedures and achieved good long term patency and limb salvage rates, well in excess of those achieved with composite prosthetic-autogenous grafts. The use of autogenous conduit appears to offer superior results to composite conduit in lower extremity revascularization. The superior durability of arm vein makes it one of the alternative conduits of choice when an adequate greater saphenous vein is not available. PMID- 10842149 TI - Infrainguinal arterial reconstructions with vein grafts in patients with prior aortic procedures: the influence of aneurysm and occlusive disease. AB - PURPOSE: This study assessed whether infrainguinal reconstructions with autogenous vein (IR) performed in patients with prior abdominal aortic aneurysm (AAA) repairs have altered graft patency, compared with those in patients who have undergone prior aortobifemoral bypass grafting procedures (ABF) for aortoiliac occlusive disease. METHODS: From 1979 to 1998, 54 patients with prior aortic reconstructions underwent 64 autogenous single-segment saphenous IRs solely for infrainguinal occlusive disease. Included in this cohort were 30 IRs with an earlier AAA repair and 34 IRs with an earlier ABF repair. During the same period, 1274 patients underwent 1642 autogenous vein lower-extremity bypass grafting procedures (LEB). Lower-extremity native arterial (AAA, n = 6; ABF, n = 11) and vein graft diameters (AAA, n = 6; ABF, n = 6) were determined by means of angiography and duplex ultrasonography, respectively. The three reconstruction groups (AAA, ABF, LEB) were compared. RESULTS: The patients in the three groups were similar in sex, indication for operation, proximal and distal anastomotic site, and number of distal runoff vessels. The cumulative 5-year primary graft patency rate in the AAA group (92% +/- 5%) was significantly higher (P <. 001) than that in the LEB group (63% +/- 2%) and the ABF group (44% +/- 11%). Furthermore, cumulative 5-year primary patency was decreased in the ABF group compared with the LEB group (P =.05). A significant increase in both native arterial (P =.001) and vein graft diameter (P <.05) was demonstrated by using linear regression and a Student t test, respectively, in the AAA group compared with the ABF group. CONCLUSION: These data demonstrate that, compared with those in patients without a previous aortic procedure, IRs in patients with prior AAA repairs have significantly improved graft patency, and IRs in patients with prior ABF reconstructions for aortoiliac occlusive disease have significantly decreased graft patency. Larger arterial diameter and altered vein graft adaptation may contribute to the superior long-term outcomes of IRs in patients with prior AAA repairs. PMID- 10842151 TI - Hand-assisted laparoscopic aortobifemoral bypass grafting. AB - OBJECTIVE: Aortobifemoral bypass grafting is a durable operation for arterial reconstruction in patients with symptomatic aortoiliac occlusive disease. In several small laparoscopic series technically demanding aortic operations have been described that have not gained widespread acceptance or applicability. To simplify the laparoscopic approach to the aorta, we have developed a technique of aortobifemoral bypass grafting that uses hand-assisted laparoscopic surgery (HALS) to minimize the complexity of aortic dissection and reconstruction. METHODS: Five patients with symptomatic aortoiliac occlusive disease underwent successful HALS aortobifemoral bypass grafting. With the use of a specialized sleeve device (Hand-Port), an operative hand was introduced into the laparoscopic field while pneumoperitoneum was maintained. Laparoscopic dissection of the infrarenal aorta was then performed with retraction provided by the operative hand. Proximal aortic anastomosis was performed with an open technique through the same 7.5-cm Hand-Port incision, and femoral anastomoses were performed in the standard fashion. RESULTS: Five hand-assisted laparoscopic aortobifemoral bypass grafts were performed (two end-to-end, three end-to-side proximal anastomoses). Mean operative time was 231 minutes. Mean blood loss was 440 mL. All patients underwent extubation immediately after surgery, were ambulatory on postoperative day (POD) 1, and were tolerating their diet by POD 3. The mean length of hospital stay was 3.8 days. One patient was discharged on POD 5 and started a clear liquid diet after a self-limiting postoperative ileus. All patients were asymptomatic and back to full activity/work by 14.6 days postoperatively, on average (range, 11-20 days). CONCLUSION: The HALS offers the advantages of tactile feedback, flexible retraction, and the introduction of conventional surgical instruments, all of which extend laparoscopic surgery and its established benefits to a wide array of more complex surgical problems, including major vascular surgery. Ease of performance, shorter hospital stays, and faster recovery times all suggest that HALS may become a valuable adjunct to conventional aortobifemoral bypass grafting. PMID- 10842150 TI - Utility and reliability of endovascular aortouniiliac with femorofemoral crossover graft for aortoiliac aneurysmal disease. AB - OBJECTIVE: The purpose of this study was to determine the early efficacy of endovascular aortouniiliac stent grafts with femorofemoral bypass graft in the treatment of aortoiliac aneurysmal disease. METHODS: We analyzed 51 consecutive patients from January 1997 to March 1999 with a mean follow-up of 15.8 months. Patients ranged in age from 44 to 93 years (mean, 75 years) with a mean aortic aneurysm diameter of 6.2 cm. Technical success was achieved in 50 patients; one patient required conversion to open repair intraoperatively. We placed 28 custom made and 22 commercial devices. The mean operative time was 223 minutes. The endograft was extended to the external iliac artery in 42% of cases. The contralateral common iliac artery was occluded using either a closed covered stent or intraluminal coils. RESULTS: The median hospital stay was 4 days with an average intensive care unit stay of 0.25 days. There were no operative mortalities. Two patients died during follow-up from unrelated conditions. Endoleaks occurred in 11 patients (22%); seven patients (14%) required intervention (four catheter based, three operative). Other complications occurred in 38% of patients but were largely remote or wound related. One femorofemoral bypass graft occluded immediately postoperatively as a result of an intraprocedural external iliac dissection yielding a 98% primary patency and 100% secondary patency. Clinical success was achieved in 88% of patients. CONCLUSIONS: These data suggest that this strategy represents a reliable method of repair of aortoiliac aneurysmal disease and extends the capability of an endoluminal approach to patients with complex iliac anatomy. PMID- 10842152 TI - Gene delivery to in situ veins: differential effects of adenovirus and adeno associated viral vectors. AB - PURPOSE: Gene transfer offers the potential to modify vein graft biology at the time of surgical implantation. Efficiency of gene delivery, stability of expression, and host responses are critical parameters for candidate vectors. We compared the effects of intraluminal exposure with adenovirus (AD) and adeno associated virus (AAV) vectors on transgene expression and monocyte adhesion (MA) in treated vein segments. METHODS: Adult New Zealand white rabbits (N = 51) were anesthetized, and the jugular veins were cannulated bilaterally. Veins were gently distended with either vector (2.10(8) to 1.10(10) infective particles/mL) or vehicle (control) for 30 minutes, after which venous flow was restored. AD and AAV vectors encoding for the marker genes beta-galactosidase (LacZ) and green fluorescent protein (GFP) were used. Vessels were explanted 2 to 40 days postinfection for analysis of gene expression (X-gal staining, reverse transcriptase-polymerase chain reaction), MA, and immunohistochemistry. Ex vivo adhesion assays used (51)Cr-labeled THP-1 cells. Statistical significance was tested by using analysis of variance with a P value less than.05. RESULTS: All animals survived, and all treated veins were patent at sacrifice. Intraluminal exposure to AD at a titer of 1.10(9) resulted in near complete transduction of the endothelium at 2 days, with no detectable expression by day 14. At an equal titer of infectious particles, transgene expression was markedly less for AAV at 2 to 7 days, but improved at 2 weeks and persisted to 40 days. MA was significantly increased 2 days after AD exposure (2.7-fold vs control, *P <.002); AAV treatment had no discernible effect on MA. CONCLUSION: AD-mediated gene transfer to vein segments resulted in robust, transient gene expression that disappeared after 2 weeks. In comparison, AAV-mediated gene delivery was less efficient, but resulted in delayed onset, persistent expression beyond 30 days. AD exposure induced an early increase in MA to the vein surface that was not seen with AAV treatment. Current generations of both AD and AAV vectors have significant, albeit different, limitations for vascular gene therapy. PMID- 10842153 TI - Inhibition of phosphatidylinositol 3-kinase and protein kinase C attenuates extracellular matrix protein-induced vascular smooth muscle cell chemotaxis. AB - PURPOSE: Intimal hyperplasia (IH), a significant cause of vascular reconstructive failure, is characterized by abnormal vascular smooth muscle cell (VSMC) migration, proliferation, and extracellular matrix (ECM) deposition. The ECM proteins, thrombospondin-1 (TSP-1), fibronectin (Fn), and vitronectin (Vn) can induce VSMC migration; however, the cellular signaling pathways involved are not identical for each ECM protein. Phosphatidylinositol 3-kinase (PI3K) and protein kinase C (PKC) are two enzymes that have been associated with VSMC migration. We sought to elucidate the roles of these enzymes in TSP-1-, Fn-, and Vn-stimulated VSMC migration. METHODS: Chemotaxis assays were performed by using a modified Boyden Chamber. TSP-1, Fn, or Vn (20 microg/mL) or serum-free media (SFM) was placed in the bottom wells of the chamber. Quiescent bovine aortic VSMC were preincubated with LY 294002 (100 micromol/L), a PI3K inhibitor, bisindolylmaleimide I (GF 109203X, 1 micromol/L), a PKC inhibitor, or in SFM alone for 30 minutes. VSMCs (50,000 cells per well) were then placed in the top wells of the chamber, and the assay was conducted for 4 hours at 37 degrees C. Results were recorded as the number of cells migrated per five fields (400x) and analyzed by means of the paired t test, with P value less than.05 considered to be significant (n = 3). RESULTS: The VSMC migration was significantly increased by TSP-1, Fn, and Vn. LY 294002 inhibited TSP-1-, Fn-, and Vn-stimulated VSMC migration (85% to 89%, P <.05). GF 109203X inhibited only TSP-1-stimulated migration (65%, P <.05). CONCLUSION: These results suggest that TSP-1-, Fn-, and Vn-stimulated migration is at least partially dependent on PI3K. However, only TSP-1 stimulated migration is at least partially dependent on PKC. PMID- 10842155 TI - The durability of endovascular treatment of multisegment iliac occlusive disease. AB - PURPOSE: The effectiveness of endovascular treatment of multisegment iliac occlusive disease (involving two or more common and/or external iliac arteries) was determined. METHODS: All patients who underwent angioplasty or stenting of at least two separate iliac artery segments were identified. Demographic data were recorded. Technical success, hemodynamic success, and aortoiliac primary and primary-assisted patency were analyzed by using the Society for Vascular Surgery and International Society for Cardiovascular Surgery (SVS/ISCVS) criteria. Multivariate, life table analysis was used as a means of determining outcome predictors. RESULTS: Eighty-seven patients underwent 207 iliac artery angioplasties and 115 iliac artery stents, which were performed in 210 iliac segments for disabling claudication in 60% of cases, for rest pain in 17% of cases, and for tissue loss in 23% of cases. Two iliac segments were treated in 64% of patients, three segments were treated in 28% of patients, and four segments were treated in 8% of patients. The complication rate was 11%. Initial hemodynamic success was achieved in 72% of cases. Clinical improvement occurred in 88% of patients. Subsequent endovascular reintervention was required in 29% of patients, whereas surgical inflow procedures were required in 14% of patients to maintain aortoiliac patency. The mean time from the primary intervention to the first reintervention was 10 +/- 3 months. At 6, 12, and 36 months after intervention, the primary patency rates of the aortoiliac segment were 76%, 61%, and 43%, respectively, and the primary assisted patency rates were 95%, 87%, and 72%, respectively. Only the presence of an external iliac artery stenosis adversely affected both primary and assisted-primary patency. At 6, 12, and 36 months, the aortoiliac primary patency rates in patients without the presence of an external iliac artery stenosis were 88%, 78%, and 69%, respectively, compared with 68%, 47%, and 18%, respectively, in patients with external iliac artery lesions (P <. 0001). CONCLUSION: Endovascular therapy for multisegment aortoiliac occlusive disease has acceptable patency rates; however, reintervention is often needed. The presence of external iliac artery disease is a significant predictor of poor outcome. PMID- 10842154 TI - Local photodynamic action of methylene blue favorably modulates the postinterventional vascular wound healing response. AB - PURPOSE: Photodynamic therapy (PDT), the light activation of photosensitizers to produce free radicals, is known to inhibit experimental intimal hyperplasia (IH). However, its clinical application has been limited by the lack of a suitable approach and a clinically appropriate photosensitizer. The aim of this study was to determine the effectiveness of a clinical approach for PDT, while testing its ability to favorably modulate the vascular wound healing response. METHODS: Rat carotid arteries were balloon-injured (BI), and for PDT, the arteries were irradiated with thermoneutral laser light (lambda = 660 nm, 100 J/cm(2)) after the photosensitizer methylene blue (MB) was delivered locally. Control rats included BI alone and MB after BI alone. Arteries were analyzed after 2 weeks with morphometric evaluation (n = 6) and in situ hybridization for versican and procollagen type I gene expression (digitized image pixel analyses, n = 3). RESULTS: No IH developed in PDT-treated arteries (0 +/- 0 mm(2); compared with BI, 0.192 +/- 0.006 mm(2); P <.0001). The diameters remained unchanged (PDT, 0.95 +/- 0.04 mm; BI, 0.94 +/- 0.05 mm; uninjured artery, 0.91 +/- 0.06 mm). Arterial injury resulted in an increase of versican and procollagen type I messenger RNA (mRNA) in the adventitia and neointima. In the repopulating cells of the adventitia after PDT, there was a significant decrease in versican mRNA (% of positive pixels per high-power field: PDT, 1.13% +/- 0.39%; BI, 2.93% +/- 0.61%; P <.02), but not in procollagen type I mRNA. CONCLUSION: The decrease of versican mRNA expression of repopulating cells after PDT reflects favorable healing on a molecular level. Site-specific delivery of MB, a clinically appropriate photosensitizer, followed by PDT represents a suitable method to promote favorable healing after balloon intervention and further supports its role for inhibiting postinterventional restenosis. PMID- 10842156 TI - Incidence and treatment of intraoperative technical problems during endovascular repair of complex abdominal aortic aneurysms. AB - PURPOSE: The purpose of this study was to assess the incidence and management of intraoperative technical problems during endovascular repair (EVR) of complex abdominal aortic aneurysms (AAA). METHODS: From February 1995 to March 1999, 204 EVRs of nonruptured AAA were performed at our institution. One hundred seventy six patients had an in-house custom-made graft; 172 were aorto-uni-iliac grafts, and four were aortoaortic grafts. Twenty- eight patients had a bifurcated graft. One hundred fourteen patients (56%) were high risk for conventional open repair. One hundred nine patients (53%) were not suitable for most commercially available devices. RESULTS: Intraoperative technical problems occurred in 81 patients (40%). There were 37 endoleaks (27 proximal, 10 distal), 15 graft stenoses, one failure of graft deployment, two graft thromboses, three aortoiliac ruptures, five renal artery occlusions (one bilateral, four unilateral), and 18 internal iliac occlusions (five bilateral, 13 unilateral). Endovascular management of these problems was successful in 37 of the 81 patients (46%) and included 15 balloon dilatations, 21 additional stent placements, and one graft thrombectomy. Fifteen of the 81 patients (19%) had open procedures (four periaortic ligature placements, six open aneurysm repairs, three common iliac ligations, and two extra-anatomic bypass grafts). In the remaining 29 patients, the on-table problem was managed expectantly. During follow-up, two of 37 patients (5%) who were treated successfully with endovascular procedures experienced recurrence. There were five deaths (33%) among the 15 patients who underwent open procedures. CONCLUSION: Intraoperative problems occur frequently during the endovascular management of complex aneurysms. Many of these problems can be managed with additional endovascular techniques without an increased risk of recurrence or procedure-related complications. Open procedures in high-risk patients carry a high mortality rate. The team performing EVR of AAA should be skillful in advanced endovascular and open surgical procedures. PMID- 10842157 TI - Thrombolysis for experimental deep venous thrombosis maintains valvular competence and vasoreactivity. AB - PURPOSE: Thrombolysis protects the structural and functional integrity of vein wall in an experimental model of acute deep venous thrombosis (DVT) immediately after treatment, but late sequelae have not been studied. We designed experiments to compare the effects of thrombolysis and surgical thrombectomy at 4 weeks after the treatment of DVT. METHODS: DVT was produced bilaterally in male mongrel dogs by proximal and distal femoral vein ligation. Five dogs underwent sham operation. After 48 hours, the ligatures were removed, and the thrombosis was treated with either Fogarty balloon catheter thrombectomy (shear force, 60 g; n = 6) or catheter-directed urokinase infusion (4000 U/min for 90 minutes; n = 6). At 4 weeks, patency and valvular competence were determined by duplex ultrasound scanning. Thrombogenicity was studied by the measurement of radiolabeled fibrin and platelet deposition. Veins were explanted and prepared for histologic examination, scanning electron microscopy, and functional studies in organ chambers. RESULTS: All veins were patent at 1 month. Recanalized thrombus was observed histologically in four (66%) thrombectomized veins, one (17%) thrombolyzed vein, and none of the sham-operated veins (P =.04). Scanning electron microscopy demonstrated similar luminal endothelial cell loss (11%-25%) in all three groups. Platelet and fibrin depositions were not different among groups. Valvular incompetence (reflux duration, >0.5 sec) did not differ significantly in the groups (thrombectomized veins, 2 of 12 (17%); thrombolyzed veins, 0 of 12 (0%); P = NS). In organ chamber studies, endothelium-dependent relaxations to calcium ionophore, but not adenosine diphosphate, were inhibited by an antagonist of nitric oxide production after thrombectomy (P <.05, thrombectomy vs sham- and thrombolysis-treated veins). All veins relaxed to exogenous nitric oxide. CONCLUSION: Both thrombectomy and thrombolysis restored patency and achieved similar valvular competence. Surgical thrombectomy, however, resulted in more residual thrombus and contributed to changes in endothelium mediated relaxations at 4 weeks. Thrombolysis maintained both structural integrity and endothelial function. PMID- 10842158 TI - Ambulatory venous pressure revisited. AB - PURPOSE: The purpose of this study was to describe a method for measuring the deep venous pressure changes in the lower extremity and compare it with those obtained in the dorsal foot vein. METHODS: After cannulation of the posterior tibial vein, a catheter with a pressure transducer in its tip was inserted and placed at the knee joint level. The dorsal foot vein was also cannulated. Pressures were recorded simultaneously at both sites during toe stands and repeated with the probe in the upper, middle, and lower calf. RESULTS: The study was performed in 45 patients with signs and symptoms of chronic venous insufficiency. Duplex Doppler scanning and ascending and descending venography performed before pressure measurements revealed saphenous vein incompetence in 11 lower extremities, incompetent perforators in 11 extremities (eight were combined with saphenous incompetence), and marked compression of popliteal vein with plantar flexion in 28 extremities. No significant deep axial reflux was observed on duplex Doppler examination or descending venography. No morphologic outflow obstruction was detected. The mean deep pressure at the knee joint level fell during toe stands, -15% +/- 27 (SD), and the mean dorsal foot vein pressure drop was even more marked, -75% +/- 22 (SD). Although the exercise pressure in the dorsal foot vein decreased in all patients (range, 13-90% drop), the popliteal vein pressure increased (4-72%) in nine limbs, decreased only marginally if at all in 15 limbs (0-15%), and fell more markedly in 21 extremities (22-65%). Deep vein recovery time was considerably shorter overall as compared with the findings by the dorsal vein measurement. In the comparison of limbs with and without superficial reflux, the recovery times in the deep system were significantly shorter in limbs with superficial incompetence. CONCLUSION: Ambulatory dorsal foot venous pressure is not always accurate in detecting changes in the pressure of the tibial and popliteal veins. Although dorsal foot venous pressure may be normal, deep venous pressure may decrease to a lesser degree or even increase. PMID- 10842160 TI - Transarterial wall oxygen gradients at a prosthetic vascular graft to artery anastomosis in the rabbit. AB - PURPOSE: Artery-wall hypoxia has been proposed to contribute to many kinds of artery-wall pathologic conditions, including atherosclerosis and intimal hyperplasia. Intimal hyperplasia is common at sites of arterial injury, including an anastomosis. The purpose of this study was to determine the effect of a prosthetic vascular graft (PVG)-to-artery anastomosis on the delivery of oxygen to the artery wall. METHODS: The transarterial wall oxygen gradient in the infrarenal aorta of New Zealand White rabbits 2 mm distal to a PVG-to-artery anastomosis was measured with an oxygen microelectrode. RESULTS: Oxygen tensions were significantly decreased in the outer artery wall immediately after the creation of the anastomosis and showed a further decrease in oxygen tensions at days 7 and 14, which correlated with the absence of a vasa vasorum. After day 14, the oxygen tensions gradually increased, returning to normal by postanastomosis day 42, correlating with a return of the vasa vasorum. These changes were noted without differences in blood pressure or arterial blood oxygen concentrations. CONCLUSION: The delivery of oxygen to the artery wall is altered by the creation of a PVG-to-artery anastomosis. Low arterial oxygen tensions at a PVG-to-artery anastomosis support a role for artery-wall hypoxia in the formation of intimal hyperplasia at the site of a PVG-to-artery anastomosis. PMID- 10842159 TI - Inducible nitric oxide synthase (iNOS) expression upregulates p21 and inhibits vascular smooth muscle cell proliferation through p42/44 mitogen-activated protein kinase activation and independent of p53 and cyclic guanosine monophosphate. AB - OBJECTIVE: Overexpression of the inducible nitric oxide synthase (iNOS) gene inhibits neointimal hyperplasia after arterial injury. The purpose of this study was to examine the mechanism by which nitric oxide (NO) inhibits vascular smooth muscle cell (VSMC) proliferation, specifically focusing on signaling pathways known to be activated by NO, including cyclic guanosine monophosphate (cGMP), p53, and p42/44 mitogen-activated protein kinase (MAPK). METHODS AND RESULTS: VSMCs that were subjected to iNOS gene transfer demonstrated a reduction in proliferation (80%) that was associated with a marked increase in p21 expression. The antiproliferative and p21 stimulatory effects of NO were not suppressed by the soluble guanylate cyclase inhibitor ODQ, implicating cGMP-independent signaling. The role of p53 in NO-mediated upregulation of p21 and inhibition of proliferation was evaluated using p53 -/- VSMCs. A similar reduction in cellular proliferation and upregulation of p21 expression were achieved with iNOS gene transfer as well as treatment with the NO-donor S-nitroso-N-acetylpenicillamine (SNAP), demonstrating the p53-independent nature of these NO-mediated pathways. The transfer of the iNOS gene activated the p42/44 MAPK, and inhibition of this MAPK pathway with PD98059 partially blocked the antiproliferative effects of NO and completely inhibited the p21 stimulatory effects of NO. For confirmation that iNOS overexpression upregulated p21 in vivo, injured rat carotid arteries were infected with an adenoviral vector carrying the iNOS gene and demonstrated a marked upregulation of p21 expression at three days. However, the ability of NO to inhibit VSMC proliferation does not solely depend on p21 upregulation since the NO-donor SNAP-inhibited VSMC proliferation in p21 -/- VSMCs. CONCLUSION: Nitric oxide inhibits VSMC proliferation in association with the upregulation of p21; both occur independent of p53 and cGMP while being partially mediated through the p42/44 MAPK signaling cascade. This represents one potential mechanism by which NO inhibits VSMC proliferation. PMID- 10842161 TI - Retroperitoneal endoscopic ligation of lumbar and inferior mesenteric arteries as a treatment of persistent endoleak after endoluminal aortic aneurysm repair. AB - A 74-year-old man receiving long-term anticoagulation therapy for intermittent atrial fibrillation had a type II endoleak after endovascular abdominal aortic aneurysm repair. During an 8-month follow-up, the endoleak persisted, and the aneurysm failed to decrease in diameter. By means of a left flank retroperitoneal endoscopic surgical approach, the aneurysm was dissected free, and the lumbar arteries emanating from the aneurysm, as well as the inferior mesenteric artery, were ligated with titanium clips. A postoperative spiral computed tomography scan depicted one pair of unclipped lumbar arteries just proximal to the aortic bifurcation. After immediate reoperation with the same approach, complete thrombosis of the aneurysm sac was radiographically confirmed. PMID- 10842162 TI - Chronic ergot toxicity: A rare cause of lower extremity ischemia. AB - Chronic ergot toxicity is a rare cause of lower extremity ischemia. The cornerstone of therapy in ergot toxicity is to discontinue the use of caffeine, cigarettes, and all ergot-containing medications. Although multiple different therapies have been recommended for acute toxicity, no specific treatment is uniformly recommended in chronic toxicity. We present a case of long-term ergot use for migraine headaches in a woman who had severe chronic lower extremity claudication. This case demonstrates the unique features associated with the diagnosis and management of chronic ergot toxicity. We recommend a conservative approach consisting of observation, antiplatelet agents, and the discontinuance of ergots. If symptoms progress to rest pain or gangrene, surgical treatment should be considered. PMID- 10842163 TI - Visceral aneurysms in Ehlers-Danlos syndrome: case report and review of the literature. AB - A patient with Ehlers-Danlos syndrome type IV had a celiac artery aneurysm. There are only nine previously published cases of visceral artery aneurysms in this condition, and surgical management was used in only four of them. This is, we believe, the first published report of a successfully treated celiac artery aneurysm in this condition in which the hazards of treatment and our rationale are explained. The literature of this rare problem is also reviewed. PMID- 10842164 TI - Failure to disclose competitive interests. PMID- 10842165 TI - Venous severity scoring: An adjunct to venous outcome assessment. AB - Some measure of disease severity is needed to properly compare the outcomes of the various approaches to the treatment of chronic venous insufficiency. Comparing the outcomes of two or more different treatments in a clinical trial, or the same treatment in two or more reports from the literature cannot be done with confidence unless the relative severity of the venous disease in each treatment group is known. The CEAP (Clinical-Etiology-Anatomic-Pathophysiologic) system is an excellent classification scheme, but it cannot serve the purpose of venous severity scoring because many of its components are relatively static and others use detailed alphabetical designations. A disease severity scoring scheme needs to be quantifiable, with gradable elements that can change in response to treatment. However, an American Venous Forum committee on venous outcomes assessment has developed a venous severity scoring system based on the best usable elements of the CEAP system. Two scores are proposed. The first is a Venous Clinical Severity Score: nine clinical characteristics of chronic venous disease are graded from 0 to 3 (absent, mild, moderate, severe) with specific criteria to avoid overlap or arbitrary scoring. Zero to three points are added for differences in background conservative therapy (compression and elevation) to produce a 30 point-maximum flat scale. The second is a Venous Segmental Disease Score, which combines the Anatomic and Pathophysiologic components of CEAP. Major venous segments are graded according to presence of reflux and/or obstruction. It is entirely based on venous imaging, primarily duplex scan but also phlebographic findings. This scoring scheme weights 11 venous segments for their relative importance when involved with reflux and/or obstruction, with a maximum score of 10. A third score is simply a modification of the existing CEAP disability score that eliminates reference to work and an 8-hour working day, substituting instead the patient's prior normal activities. These new scoring schemes are intended to complement the current CEAP system. PMID- 10842166 TI - Ability of Egr1 to activate tyrosine hydroxylase transcription in PC12 cells. Cross-talk with AP-1 factors. AB - We have recently identified an Egr1 motif that overlaps with the Sp1 element in the tyrosine hydroxylase (TH) promoter. Here we examine whether this motif has a functional role in the regulation of TH transcription in PC12 cells. In nuclear extracts from control PC12 cells, an oligonucleotide containing the TH Sp1/Egr1 motif binds Sp1-containing complexes. Treatment of PC12 cells with phorbol ester (2 micrometer 12-O-tetradecanoylphorbol-13-acetate (TPA)) gives rise to a new Egr1-containing complex. TPA treatment reduces the steady-state levels of the Sp1 protein and leads to the appearance of immunoreactive Egr1 protein within 30-60 min. Expression of the Egr1 protein in PC12 cells stimulates the chloramphenicol acetyltransferase reporter gene placed under the control of the first 272 nucleotides of the rat TH promoter. Site-directed mutagenesis of either the Sp1/Egr1 motif or of an upstream AP-1 motif or both abolishes the Egr1-mediated induction of chloramphenicol acetyltransferase activity. An oligonucleotide encompassing the AP-1/E-box sequence of the rat TH promoter competes in electrophoretic mobility shift assays for binding of nuclear extracts from control and TPA-treated cells to an oligonucleotide containing the Sp1/Egr1 element, indicating that these two enhancers may interact. The results show that Egr1 can activate TH transcription and reveals cross-talk between Sp1/Egr1 and AP 1 factors. PMID- 10842167 TI - Oligomerization of mu- and delta-opioid receptors. Generation of novel functional properties. AB - The existence of dimers and oligomers for many G protein-coupled receptors has been described by us and others. Since many G protein-coupled receptor subtypes are highly homologous to each other, we examined whether closely related receptors may interact with each other directly and thus have the potential to create novel signaling units. Using mu- and delta-opioid receptors, we show that each receptor expressed individually was pharmacologically distinct and could be visualized following electrophoresis as monomers, homodimers, homotetramers, and higher molecular mass oligomers. When mu- and delta-opioid receptors were coexpressed, the highly selective synthetic agonists for each had reduced potency and altered rank order, whereas endomorphin-1 and Leu-enkephalin had enhanced affinity, suggesting the formation of a novel binding pocket. No heterodimers were visualized in the membranes coexpressing mu- and delta-receptors by the methods available. However, hetero-oligomers were identified by the ability to co immunoprecipitate mu-receptors with delta-receptors and vice versa using differentially epitope-tagged receptors. In contrast to the individually expressed mu- and delta-receptors, the coexpressed receptors showed insensitivity to pertussis toxin and continued signal transduction, likely due to interaction with a different subtype of G protein. In this study, we provide, for the first time, evidence for the direct interaction of mu- and delta-opioid receptors to form oligomers, with the generation of novel pharmacology and G protein coupling properties. PMID- 10842168 TI - The osmotic shock-induced glucose transport pathway in 3T3-L1 adipocytes is mediated by gab-1 and requires Gab-1-associated phosphatidylinositol 3-kinase activity for full activation. AB - Osmotic shock treatment of 3T3-L1 adipocytes causes an increase in glucose transport activity and translocation of GLUT4 protein similar to that elicited by insulin treatment. Insulin stimulation of GLUT4 translocation and glucose transport activity was completely inhibited by wortmannin, however, activation by osmotic shock was only partially blocked. Additionally, we have found that the newly identified insulin receptor substrate Gab-1 (Grb2-associated binder-1) is tyrosine-phosphorylated following sorbitol stimulation. Treatment of cells with the tyrosine kinase inhibitor genistein inhibited osmotic shock-stimulated Gab-1 phosphorylation as well as shock-induced glucose transport. Furthermore, pretreatment with the selective Src family kinase inhibitor PP2 completely inhibited the ability of sorbitol treatment to cause tyrosine phosphorylation of Gab-1. We have also shown that microinjection of anti-Gab-1 antibody inhibits osmotic shock-induced GLUT4 translocation. Furthermore, phosphorylated Gab-1 binds and activates phosphatidylinositol 3-kinase (PI3K) in response to osmotic shock. The PI3K activity associated with Gab-1 was 82% of that associated with anti-phosphotyrosine antibodies, indicating that Gab-1 is the major site for PI3K recruitment following osmotic shock stimulation. Although wortmannin only causes a partial block of osmotic shock-stimulated glucose uptake, wortmannin completely abolishes Gab-1 associated PI3K activity. This suggests that other tyrosine kinase-dependent pathways, in addition to the Gab-1-PI3K pathway, contribute to osmotic shock-mediated glucose transport. To date, Gab-1 is the first protein identified as a member of the osmotic shock signal transduction pathway. PMID- 10842169 TI - Rap1p-binding sites in the saccharomyces cerevisiae GPD1 promoter are involved in its response to NaCl. AB - Mechanisms involved in transcriptional regulation of the osmotically controlled GPD1 gene in Saccharomyces cerevisiae were investigated by promoter analysis. The GPD1 gene encodes NAD(+)-dependent glycerol-3-phosphate dehydrogenase, a key enzyme in the production of the compatible solute glycerol. By analysis of promoter deletions, we identified a region at nucleotides -478 to -324, in relation to start of translation, to be of great importance for both basal activity and osmotic induction of GPD1. Electrophoretic mobility shift and DNase I footprint analyses demonstrated protein binding to parts of this region that contain three consensus sequences for Rap1p (repressor activator protein 1) binding sites. Actual binding of Rap1p to this region was confirmed by demonstrating enhanced electrophoretic mobility of the protein-DNA complex with extracts containing an N-terminally truncated version of Rap1p. The detected Rap1p-DNA interactions were not affected by changes in the osmolarity of the growth medium. Specific inactivation of the Rap1p-binding sites by a C-to-A point mutation in the core of the consensus showed that this factor is a major determinant of GPD1 expression since mutations in all three putative binding sites for Rap1p strongly hampered osmotic induction and drastically lowered basal activity. We also show that the Rap1p-binding sites appear functionally distinct; the most distal site (core of the consensus at position -386) exhibited the highest affinity for Rap1p and was strictly required for low salt induction (< or =0.6 m NaCl), but not for the response at higher salinities (> or =0.8 m NaCl). This indicates tha different molecular mechanisms might be operational for low and high salt responses of the GPD1 promoter. PMID- 10842170 TI - Functional assembly of fragments from bisected smooth muscle myosin light chain kinase. AB - The C-terminal regulatory segment of smooth muscle myosin light chain kinase folds back on its catalytic core to inhibit kinase activity. This regulatory segment consists of autoinhibitory residues linking the catalytic core to the calmodulin-binding sequence and perhaps additional C-terminal residues including an immunoglobulin-like motif. However, mutational and biochemical analyses showed no specific involvement of residues C-terminal to the calmodulin-binding sequence. To obtain additional insights on the proposed mechanisms for autoinhibition and Ca(2+)/calmodulin activation of the kinase, the polypeptide backbone chain of myosin light chain kinase was cleaved by genetic means to produce N- and C-terminal protein fragments. The N-terminal fragment containing the catalytic core was catalytically inactive when expressed alone. Co-expression of the N-terminal fragment with the C-terminal fragment containing the regulatory segment restored kinase activity. Deletion of the autoinhibitory linker residues without or with the calmodulin-binding sequence prevented restoration of kinase activity. In the presence or absence of Ca(2+)/calmodulin, regulatory segment binding occurred through the linker region connecting the catalytic core to the calmodulin-binding sequence. Collectively, these results indicate that residues C terminal to the calmodulin-binding sequence (including the immunoglobulin-like motif) are not functional components of the regulatory segment. Furthermore, the principal autoinhibitory motif is contained in the sequence linking the catalytic core of myosin light chain kinase to the calmodulin-binding sequence. PMID- 10842171 TI - The conformation of calreticulin is influenced by the endoplasmic reticulum luminal environment. AB - In order to understand the dynamics of the endoplasmic reticulum (ER) luminal environment, we investigated the role of Ca(2+), Zn(2+), and ATP on conformational changes of calreticulin. Purified calreticulin was digested with trypsin in the presence or absence of Ca(2+), Zn(2+), and ATP. At low Ca(2+) concentration (<100 micrometer), calreticulin is rapidly and fully degraded by trypsin, indicating that under these conditions the protein is in a highly trypsin-susceptible conformation. Increasing Ca(2+) concentration up to 500 micrometer or 1 mm resulted in protection of the full-length calreticulin and in generation of the 27-kDa fragment highly resistant to trypsin digestion. The 27 kDa protease-resistant core of the protein represented the NH(2)-terminal half of calreticulin and was identified by its reactivity with specific antibodies and by NH(2)-terminal amino acid sequence analysis. Ca(2+)-dependent changes in calreticulin's sensitivity to proteolysis indicate that agonist-induced fluctuation in the free ER luminal Ca(2+) concentration may affect the protein conformation and function. Trypsin digestion of calreticulin in the presence of Zn(2+) resulted in the formation of a 17-kDa central protease-resistant core in the protein corresponding to the central region of the protein, indicating that under these conditions the N- and C-domains of the protein are in an extended conformation. Here we also show that calreticulin is an ATP-binding protein but that it does not contain detectable ATPase activity. Digestion of the protein with trypsin in the presence of Mg(2+)-ATP protects the full-length protein. These results indicate that calreticulin may undergo frequent, ion-induced conformation changes, which may affect its function and its ability to interact with other proteins in the lumen of the ER. PMID- 10842172 TI - Hyperosmotic stress stimulates promoter activity and regulates cellular utilization of the serum- and glucocorticoid-inducible protein kinase (Sgk) by a p38 MAPK-dependent pathway. AB - We have established that the serum- and glucocorticoid-inducible protein kinase (Sgk) is a new component of the hyperosmotic stress response. Treatment of NMuMg mammary epithelial cells with the organic osmolyte, sorbitol, caused the stable accumulation of Sgk transcripts and protein after an approximately 4-h lag. Transient transfection of a series of sgk-CAT reporter plasmids containing either 5' deletions or continuous 6-base pair substitutions identified a hyperosmotic stress-regulated element that is GC-rich and is necessary for the sorbitol stimulation of sgk gene promoter activity. Gel shift analysis identified four major DNA-protein complexes in the hyperosmotic stress-regulated element that, by competition with excess consensus wild type and mutant oligonucleotides and by antibody supershifts, contains the Sp1 transcription factor. Several lines of evidence suggest that the p38 MAPK signaling pathway mediates the hyperosmotic stress stimulation of sgk gene expression. Treatment with pharmacological inhibitors of p38 MAPK or with a dominant negative form of MKK3, an upstream regulator of p38 MAPK, significantly reduced or ablated the sorbitol induction of sgk promoter activity or protein production. Using an in vitro peptide transphosphorylation assay, sorbitol treatment activates either endogenous or exogenous Sgk that is localized to the cytoplasmic compartment. Thus, we propose that the stimulated expression of enzymatically active Sgk after sorbitol treatment is a newly defined component of the p38 MAPK-mediated response to hyperosmotic stress. PMID- 10842173 TI - Structure/function of the human Ga1beta1,3-glucuronosyltransferase. Dimerization and functional activity are mediated by two crucial cysteine residues. AB - Galbeta1,3-glucuronosyltransferase (GlcAT-I) that catalyzes the transfer of a glucuronic acid residue onto the trisaccharide primer of the glycosaminoglycan protein linkage region plays an essential role in the early steps of the biosynthesis of glycosaminoglycans. In order to gain insight into the structure/function of the enzyme, the human recombinant GlcAT-I was successfully expressed in the yeast Pichia pastoris, with an apparent molecular mass of 43 kDa. Analysis of the electrophoretic mobility of the membrane-bound protein in nonreducing and reducing conditions, together with cross-linking studies, indicated that the membrane-bound GlcAT-I formed active disulfide-linked dimers. GlcAT-I expressed without the predicted N-terminal cytoplasmic tail or secreted as a polypeptide lacking the cytoplasmic tail and transmembrane domain was similarly organized as dimers, suggesting that the structural determinants for the dimerization state are localized in the luminal domain of the protein. In addition, the role of Cys(33) and Cys(301) in that process was investigated by site-directed mutagenesis combined with chemical modification of GlcAT-I by N phenylmaleimide. Replacement of Cys(33) with alanine abolished the formation of dimers with a concomitant decrease in the catalytic efficiency mainly due to a decrease in apparent maximal velocity and in affinity for UDP-glucuronic acid. On the other hand, N-phenylmaleimide treatment or alanine substitution of the Cys(301) residue inactivated the enzyme. Our study demonstrates that GlcAT-I is organized as a homodimer as a result of disulfide bond formation mediated by Cys(33) localized in the stem region, whereas the residue Cys(301) localized in a conserved C-terminal domain is strictly required for the functional integrity of the enzyme. PMID- 10842174 TI - myo-inositol 3,4,5,6-tetrakisphosphate inhibits an apical calcium-activated chloride conductance in polarized monolayers of a cystic fibrosis cell line. AB - Does inositol 3,4,5,6-tetrakisphosphate (Ins(3,4,5,6)P(4)) inhibit apical Ca(2+) activated Cl(-) conductance (CaCC)? We studied this question using human CFPAC-1 pancreatoma cells grown in polarized monolayers. Cellular Ins(3,4,5,6)P(4) levels were acutely sensitive to purinergic receptor activation, rising 3-fold within 1 min of agonist addition. Intracellular Ins(3,4,5,6)P(4) levels were therefore specifically elevated, independently of receptor activation, by incubating cells with a cell-permeant bioactivable analogue, 1,2-di-O-butyl-myo-inositol 3,4,5,6 tetrakisphosphate octakis(acetoxymethyl)ester (Bt(2)Ins (3,4,5,6)P(4)/AM). The latter inhibited Ca(2+)-activated Cl(-) secretion by 60%. We next used nystatin to selectively permeabilize the basolateral membrane to monovalent anions and cations, thereby preventing this membrane from electrochemically dominating ion movements through the apical membrane. Thus, we studied autonomous regulation of apical Cl(-) channels in situ. The properties of Cl(-) flux across the apical membrane were those expected of CaCC: niflumic acid sensitivity, outward rectification, and 2-fold greater permeability of I(-) over Cl(-). Following nystatin-treatment, we elevated intracellular levels of Ins(3,4,5,6)P(4) with either purinergic agonists or with Bt(2)Ins(3,4,5,6)P(4)/AM. Both protocols inhibited Ca(2+)-activated Cl(-) secretion (up to 70%). These studies provide the first demonstration that, in a physiologically relevant context of a polarized monolayer, there is an apical, Ins(3,4,5,6)P(4)-inhibited CaCC. PMID- 10842175 TI - Distant enhancers stimulate the albumin promoter through complex proximal binding sites. AB - The albumin-alpha-fetoprotein locus epitomizes the main features of transcriptional regulation of fetal and adult hepatocyte-specific genes: developmentally regulated promoters and strong distant enhancers. Full enhancer activity required only a proximal albumin-promoter region containing the TATA box, hepatic nuclear factor 1 (HNF1), and nuclear factor Y (NF-Y) sites. Deletion of the HNF1 site abrogated enhancer and promoter activity, whereas methylation of the site reduced all activity by about 3-fold. Deletion of the NF-Y site attenuated activity by about half, but much of the activity could be replaced by juxtaposition of an upstream region (designated distal element IV). Gel shift and competition analysis demonstrated that binding of architectural factors overlapped NF-Y binding. Moreover, a mutation that eliminated NF-Y binding but only minimally perturbed the surrounding region did not affect enhancer function. In plasmids with a second promoter, the enhancers simultaneously stimulated both albumin and alpha-fetoprotein promoters with minimal competition, but surprisingly some mutations in the albumin promoter attenuated expression from both promoters, whereas another uncoupled their expression. With single promoters, the function of the proximal promoter region was controlled by three parameters in the following hierarchy: HNF1 binding > local architecture > NF-Y binding, but integrated two-promoter function had a much greater dependence on NF Y. PMID- 10842176 TI - Oncogenic Ras induces p19ARF and growth arrest in mouse embryo fibroblasts lacking p21Cip1 and p27Kip1 without activating cyclin D-dependent kinases. AB - Oncogenic Ras induces two products of the INK4a/ARF tumor suppressor locus (p16(INK4a) and p19(ARF)) in primary human and rodent fibroblasts, ultimately leading to a permanent state of cell cycle arrest resembling replicative senescence. Whereas p16(INK4a) antagonizes the activities of cyclin D-dependent kinases, p19(ARF) activates the p53 transcription factor. Immortalized rodent fibroblast cell lines that lack INK4a/ARF function, ARF alone, or p53 are resistant to the growth inhibitory effects of oncogenic Ras and instead continue to proliferate and undergo morphological transformation. Primary mouse embryo fibroblasts lacking Cip1 and Kip1 genes encoding inhibitors of cyclin-dependent kinase-2 were used to further explore the effects of oncogenic Ras on arrest of the cell division cycle. Although early passage primary fibroblast strains that lack both p21(Cip1) and p27(Kip1) fail to assemble cyclin D-dependent kinases, oncogenic Ras retained its ability to induce p19(ARF), but not p16(INK4a), protecting Cip/Kip-null cells from proliferating and undergoing transformation. Under these conditions, Ras did not induce G(1) phase arrest but instead triggered DNA synthesis, abnormal nuclear divisions, failure of cytokinesis, and emergence of polyploid cells. Therefore, in the absence of p16(INK4a), p21(Cip1), and p27(Kip1), oncogenic Ras affects the functions of genes required for completion of the cell cycle. PMID- 10842177 TI - Phosphorylation state of HPr determines the level of expression and the extent of phosphorylation of the lactose transport protein of Streptococcus thermophilus. AB - The lactose transport protein (LacS) of Streptococcus thermophilus is composed of a translocator domain and a regulatory domain that is phosphorylated by HPr(His approximately P), the general energy coupling protein of the phosphoenolpyruvate:carbohydrate phosphotransferase system (PTS). Lactose transport is affected by the phosphorylation state of HPr through changes in the activity of the LacS protein as well as expression of the lacS gene. To address whether or not CcpA-HPr(Ser-P)-mediated catabolite control is involved, the levels of LacS were determined under conditions in which the cellular phosphorylation state of HPr greatly differed. It appears that HPr(Ser-P) is mainly present in the exponential phase of growth, whereas HPr(His approximately P) dominates in the stationary phase. The transition from HPr(Ser-P) to HPr(His approximately P) parallels an increase in LacS level, a drop in lactose and an increase in galactose concentration in the growth medium. Because the K(m)(out) for lactose is higher than that for galactose, the lactose transport capacity decreases as lactose concentration decreases and galactose accumulates in the medium. Our data indicate that S. thermophilus compensates for the diminished transport capacity by synthesizing more LacS and phosphorylating the protein, which results in increased transport activity. The link between transport capacity and lacS expression levels and LacS phosphorylation are discussed. PMID- 10842178 TI - HPr(His approximately P)-mediated phosphorylation differently affects counterflow and proton motive force-driven uptake via the lactose transport protein of Streptococcus thermophilus. AB - The lactose transport protein (LacS) of Streptococcus thermophilus has a C terminal hydrophilic domain that is homologous to IIA protein and protein domains of the phosphoenolpyruvate:carbohydrate phosphotransferase system (PTS). The IIA domain of LacS is phosphorylated on His-552 by the general energy coupling proteins of the PTS, which are Enzyme I and HPr. To study the effect of phosphorylation on transport, the LacS protein was purified and incorporated into liposomes with the IIA domain facing outwards. This allowed the phosphorylation of the membrane-reconstituted protein by purified HPr(His approximately P) of S. thermophilus. Phosphorylation of LacS increased the V(max) of counterflow transport, whereas the V(max) of the proton motive force (delta p)-driven lactose uptake was not affected. In line with a range of kinetic studies, we propose that phosphorylation affects the rate constants for the reorientation of the ternary complex (LacS with bound lactose plus proton), which is rate-determining for counterflow but not for delta p-driven transport. PMID- 10842179 TI - Selective elimination of high constitutive activity or chemokine binding in the human herpesvirus 8 encoded seven transmembrane oncogene ORF74. AB - Open reading frame 74 (ORF74) encoded by human herpesvirus 8 is a highly constitutively active seven transmembrane (7TM) receptor stimulated by angiogenic chemokines, e.g. growth-related oncogene-alpha, and inhibited by angiostatic chemokines e.g. interferon-gamma-inducible protein. Transgenic mice expressing ORF74 under control of the CD2 promoter develop highly vascularized Kaposi's sarcoma-like tumors. Through targeted mutagenesis we here create three distinct phenotypes of ORF74: a receptor with normal, high constitutive signaling through the phospholipase C pathway but deprived of binding and action of chemokines obtained through deletion of 22 amino acids from the N-terminal extension; an ORF74 with high constitutive activity but with selective elimination of stimulatory regulation by angiogenic chemokines obtained through substitution of basic residues at the extracellular ends of TM-V or TM-VI; and an ORF74 lacking constitutive activity but with preserved ability to be stimulated by agonist chemokines obtained through introduction of an Asp residue on the hydrophobic, presumed membrane-exposed face of TM-II. It is concluded that careful molecular dissection can selectively eliminate either agonist or inverse agonist modulation as well as high constitutive activity of the virally encoded oncogene ORF74 and that these mutant forms presumably can be used in transgenic animals to identify the molecular mechanism of its transforming activity. PMID- 10842180 TI - Doppel is an N-glycosylated, glycosylphosphatidylinositol-anchored protein. Expression in testis and ectopic production in the brains of Prnp(0/0) mice predisposed to Purkinje cell loss. AB - The Prnd gene encodes a homolog of the cellular prion protein (PrP(C)) called doppel (Dpl). Up-regulation of Prnd mRNA in two distinct lines of PrP gene ablated (Prnp(0/0)) mice, designated Rcm0 and Ngsk, is associated with death of Purkinje cells. Using recombinant Dpl expressed in Escherichia coli and mouse neuroblastoma cells we demonstrate that wild type (wt) Dpl, like PrP(C), adopts a predominantly alpha-helical conformation, forms intramolecular disulfide bonds, has two N-linked oligosaccharides, and is presented on the cell surface via a glycosylphosphatidylinositol anchor. Dpl protein was detected in testis of wt mice. Using Triton X-114 phase partitioning to enrich for glycosylphosphatidylinositol-anchored proteins, Dpl was detected in brain samples from Rcm0 Prnp(0/0) mice but was absent in equivalent samples from wt mice and ZrchI Prnp(0/0) mice, indicating that ectopic expression of this protein may cause cerebellar pathology in Rcm0 mice. Biochemical and structural similarities between PrP(C) and Dpl documented here parallel the observation that ataxic Ngsk Prnp(0/0) mice can be rescued by overexpression of wild-type PrP transgenes, and suggest that cell surface PrP(C) can antagonize the toxic effect of Dpl expressed in the central nervous system. PMID- 10842181 TI - The interaction of neuropilin-1 with vascular endothelial growth factor and its receptor flt-1. AB - Neuropilin-1 (NP-1) was first identified as a semaphorin receptor involved in neuron guidance. Subsequent studies demonstrated that NP-1 also binds an isoform of vascular endothelial growth factor (VEGF) as well as several VEGF homologs, suggesting that NP-1 may also function in angiogenesis. Here we report in vitro binding experiments that shed light on the interaction between VEGF165 and NP-1, as well as a previously unknown interaction between NP-1 and one of the VEGF receptor tyrosine kinases, VEGFR1 or Flt-1. BIAcore analysis demonstrated that, with the extracellular domain (ECD) of NP-1 immobilized at low density, VEGF165 bound with low affinity (K(d) = 2 microm) and fast kinetics. The interaction was dependent on the heparin-binding domain of VEGF165 and increased the affinity of VEGF165 for its signaling receptor VEGFR2 or kinase insert domain-containing receptor. The affinity of VEGF165 for the NP-1 ECD was greatly enhanced either by increasing the density of immobilized NP-1 (K(d) = 113 nm) or by the addition of heparin (K(d) = 25 nm). We attribute these affinity enhancements to avidity effects mediated by the bivalent VEGF165 homodimer or multivalent heparin. We also show that the NP-1 ECD binds with high affinity (K(d) = 1.8 nm) to domains 3 and 4 of Flt-1 and that this interaction inhibits the binding of NP-1 to VEGF165. Based on these results, we propose that NP-1 acts as a coreceptor for various ligands and that these functions are dependent on the density of NP-1 on the cell membrane. Furthermore, Flt-1 may function as a negative regulator of angiogenesis by competing for NP-1. PMID- 10842182 TI - 3'-Azidothymidine potently inhibits the biosynthesis of highly branched N-linked oligosaccharides and poly-N-acetyllactosamine chains in cells. AB - Previous studies in our laboratory have characterized 3'-azido-3'-deoxythymidine (AZT) as a potent inhibitor of glycosphingolipid biosynthesis in cultured cells (Steet, R., Alizadeh, M., Melancon, P., and Kuchta, R. D. (1999) Glycoconj. J. 16, 237-245; Yan, J.-P., Ilsley, D. D., Frohlick, C., Steet, R., Hall, E. T., Kuchta, R. D., and Melancon, P. (1995) J. Biol. Chem. 270, 22836-22841). Here, we report that AZT treatment of K562 cells results in significant alterations in the profile of N-linked oligosaccharides. Fractionation of [(3)H]mannose-labeled oligosaccharides from AZT-treated K562 cells using lectin affinity chromatography revealed striking changes in the branching and processing of N-linked glycoconjugates. AZT treatment resulted in the production of fewer highly branched complex glycans (60% of control at 20 micrometer AZT) and a significant accumulation of core-fucosylated biantennary oligosaccharides. In addition, extension of branched oligosaccharides with multiple poly-N-acetyllactosamine repeats is nearly abolished by AZT concentrations as low as 2 micrometer. A shift from multiantennary to moderately branched oligosaccharides was also apparent in the melanoma cell line SK-MEL-30 upon AZT treatment. N-Linked glycans from both cell lines exhibited increased affinity for the beta-galactoside-binding lectin RCA-I in the presence of AZT, suggesting that the addition of terminal sialic acid is sensitive to the drug. These results demonstrate the ability of AZT to modulate strongly the processing of asparagine-linked glycoconjugates in whole cells and reveal a novel mechanism by which AZT treatment may cause anemia. PMID- 10842183 TI - Mammalian ASIC2a and ASIC3 subunits co-assemble into heteromeric proton-gated channels sensitive to Gd3+. AB - Proton receptors of the acid-sensing ion channel (ASIC) family are expressed in sensory neurons and thus could play a critical role in the detection of noxious acidosis. To investigate the subunit composition of native ASICs in peripheral and central neurons, we co-injected human as well as rodent ASIC2a and ASIC3 subunits in Xenopus oocytes. The amplitudes of acid-induced biphasic responses mediated by co-expressed ASIC2a and ASIC3 subunits were much larger (as much as 20-fold) than the currents mediated by the respective homomers, clearly indicating functional association. The reversal potential of the ASIC2a+3 current (>/=+20 mV) reflected a cationic current mainly selective for sodium. The sensitivity to pH or amiloride of single versus co-expressed ASIC subunits was not significantly different; however, gadolinium ions inhibited ASIC3 and ASIC2a+3 responses with much higher potency (IC(50) approximately 40 microm) than the ASIC2a response (IC(50) >/=1 mm). Biochemical interaction between ASIC2a and ASIC3 subunits was demonstrated by co-purification from transfected human embryonic kidney (HEK293) cells and Xenopus oocytes. Our in situ hybridization data showed that rat ASIC2a and ASIC3 transcripts are co-localized centrally, whereas reverse transcription-polymerase chain reaction data led us to detect co expression of human ASIC2a and ASIC3 subunits in trigeminal sensory ganglia, brain, and testis where they might co-assemble into a novel subtype of proton gated channels sensitive to gadolinium. PMID- 10842184 TI - Negative regulation of growth hormone receptor/JAK2 signaling by signal regulatory protein alpha. AB - Signal regulatory proteins (SIRPs) are receptor-like transmembrane proteins, the majority of which contain a cytoplasmic proline-rich region and four cytoplasmic tyrosines that, when phosphorylated, bind SH2 domain-containing protein tyrosine phosphatases (SHP). We demonstrated previously that growth hormone (GH) induces tyrosyl phosphorylation of SIRPalpha and association of SIRPalpha with SHP-2. The GH-activated tyrosine kinase JAK2 associates with and tyrosyl-phosphorylates SIRPalpha1. Here we show that JAK2-SIRPalpha1 association does not require phosphotyrosines in SIRPalpha1 or JAK2 or the proline-rich region of SIRPalpha1. However, when the C-terminal 30 amino acids of SIRPalpha1 containing the proline rich region and tyrosine 495 are deleted, tyrosyl phosphorylation of SIRPalpha1 by JAK2 and association of SHP-2 with SIRPalpha1 are reduced. GH-dependent tyrosyl phosphorylation of JAK2 is reduced when wild-type SIRPalpha1 compared with SIRPalpha1 lacking the four cytoplasmic tyrosines (SIRP 4YF) is expressed in cells, suggesting that SIRPalpha1 negatively regulates GHR/JAK2 signaling. Consistent with reduced JAK2 activity, overexpression of wild-type SIRPalpha1 but not SIRP 4YF reduces GH-induced phosphorylation of ERKs 1 and 2, STAT3, and STAT5B. These results suggest that SIRPalpha1 is a negative regulator of GH signaling and that the ability of SIRPalpha1 mutants to negatively regulate GHR JAK2 signaling correlates with their ability to bind SHP-2. PMID- 10842185 TI - Hepatitis B viral transactivator HBx alleviates p53-mediated repression of alpha fetoprotein gene expression. AB - Chronic infection with hepatitis B virus (HBV) is associated with development of hepatocellular carcinoma (HCC). The exact mechanism by which chronic infection with HBV contributes to onset of HCC is unknown. However, previous studies have implicated the HBV transactivator protein, HBx, in progression of HCC through its ability to bind the human tumor suppressor protein, p53. In this study, we have examined the ability of HBx to modify p53 regulation of the HCC tumor marker gene, alpha-fetoprotein (AFP). By utilizing in vitro chromatin assembly of DNA templates prior to transcription analysis, we have demonstrated that HBx functionally disrupts p53-mediated repression of AFP transcription through protein-protein interaction. HBx modification of p53 gene regulation is both tissue-specific and dependent upon the p53 binding element. Our data suggest that the mechanism by which HBx alleviates p53 repression of AFP transcription is through an association with DNA-bound p53, resulting in a loss of p53 interaction with liver-specific transcriptional co-repressors. PMID- 10842186 TI - Ectopic expression of transcription factor NF-E2 alters the phenotype of erythroid and monoblastoid cells. AB - In this study, regulation of transcription factor NF-E2 was examined in differentiating erythroid and myeloid cells, and the impact of raising NF-E2 concentrations within these cell types was assessed. NF-E2 was expressed in the J2E erythroid cell line, but the levels increased only marginally during erythropoietin-induced differentiation. In contrast, rare myeloid variants of J2E cells did not express NF-E2. Although NF-E2 was present in M1 monoblastoid cells, it was undetectable as these cells matured into macrophages. Compared with erythroid cells, transcription of the NF-E2 gene was reduced, and the half-life of the mRNA was significantly shorter in monocytoid cells. Ectopic expression of NF-E2 had a profound impact upon the J2E cells; morphologically mature erythroid cells spontaneously emerged in culture, but the cells failed to synthesize hemoglobin, even in the presence of erythropoietin. Although proliferation and viability increased in the NF-E2-transfected J2E cells, their responsiveness to erythropoietin was severely diminished. Strikingly, increasing the expression of NF-E2 in M1 cells produced sublines that contained erythroid or immature megakaryocytic cells. Finally, overexpression of NF-E2 in primary hemopoietic progenitors from fetal liver increased erythroid colony formation in the absence of erythropoietin. These data demonstrate that elevated NF-E2 (i) had a dominant effect on the phenotype and maturation of J2E erythroid cells, (ii) was able to reprogram the M1 monocytoid line, and (iii) promoted the development of erythroid colonies by normal progenitors. PMID- 10842187 TI - Tn antigen is a pre-cancerous biomarker in breast tissue and serum in n nitrosomethylurea-induced rat mammary carcinogenesis. AB - The Tn determinant (GalNAcalpha-O-Ser/Thr), normally a cryptic structure in mucin type O-glycans, is a tumor-associated marker which has attracted particular interest in cancer biology. We herein report the characterization of N nitrosomethylurea (NMU)-induced breast cancer in rats as a new model for the study of aberrant O-glycosylation products. Tn-antigen expression is detectable not only in mammary carcinoma induced by NMU but also in carcinogen-initiated mammary epithelium, indicating that Tn could be a pre-cancerous biomarker in rats treated with NMU. Serum Tn levels were followed up longitudinally in 30 rats from the time of the first injection of NMU to the development of advanced breast cancer. Tn antigen increased in serum several weeks before tumor development, and became highly positive after 56 days of carcinogenesis (prior to breast-cancer occurrence), and the levels correlated with Tn expression in mammary tissues. However, during the follow-up after detection of mammary cancer, all animals displayed a significant decrease of serum Tn antigen, and low levels were observed in animals with advanced breast cancer. We have shown that the humoral immune response to cancer, with the production of anti-Tn antibodies, could hamper the detection of Tn antigen in animals with advanced breast cancer. These results suggest that NMU-induced rat mammary carcinogenesis is a useful experimental model to study the regulation of O-glycosylation at the cellular level during malignant transformation. PMID- 10842188 TI - Angiostatin generation by human tumor cell lines: involvement of plasminogen activators. AB - Angiostatin is a tumor-derived angiogenesis inhibitor consisting of an internal fragment of plasminogen. Little is known about the production of angiostatin by human tumors. In this study, we examined the in vitro angiostatin-generating capacities of a panel of human tumor cell lines (total n = 75) and the proteolytic molecule(s) involved. Angiostatin formation was determined by assessing the level of plasminogen digestion in conditioned medium by Western blot analysis. We found that the capacity to produce angiostatin is a common feature of many cell lines, depending on the tumor type. All 6 bladder-carcinoma and 6 out of 7 prostate-carcinoma cell lines showed intermediate to potent angiostatin-generating activity. In contrast, only 2 out of 7 colon-carcinoma and 2 out of 9 renal-cell carcinoma cell lines were able to generate angiostatin at intermediate levels. Out of 25 melanoma cell lines, only one line failed to generate angiostatin. In the other cell-line groups (cervix, breast and ovary), angiostatin formation varied. Remarkably, angiostatin bands were not of equal size in all plasminogen digests. Since reported data have indicated that plasminogen activators (uPA and tPA) were able to excise the angiostatin fragment from the plasminogen parent molecule via plasmin generation, we determined levels of uPA and tPA and PAI-1 antigen in the conditioned media, and correlated the results with angiostatin-generating capacity. Whereas prostate- and bladder carcinoma lines capable of generating high levels of angiostatin showed high uPA levels, angiostatin generation in melanoma cell lines was correlated with tPA levels. Generally, angiostatin non-producers did not express uPA or tPA. In 6 out of 75 cell lines, however, we found angiostatin generation combined with low or absent levels of plasminogen activator, suggesting the involvement of alternative proteolytic pathways in the generation of angiostatin. PMID- 10842189 TI - Angiogenic balance in human melanoma: expression of VEGF, bFGF, IL-8, PDGF and angiostatin in relation to vascular density of xenografts in vivo. AB - Tumor angiogenesis, a major requirement for tumor outgrowth and metastasis formation, is regulated by pro- and anti-angiogenic factors. We have studied the expression of a panel of angiogenic factors, and of the angiogenesis inhibitor angiostatin, in a panel of human melanoma cell lines giving rise to xenografts with different vascular densities. Angiogenic-factor expression was analyzed in vitro (cell lines) and in vivo (xenografts), both at mRNA (RT-PCR and Northern blot) and at protein level (ELISA and Western blot). In vitro angiostatin generation was assessed by Western-blot analysis. Expression of bFGF and VEGF was clearly correlated with a high degree of vascularization, confirming the importance of these factors for tumor angiogenesis. In addition, there was exclusive or elevated in vitro expression of angiogenic factors IL-8, PDGF-AB, and, to a lesser extent, midkine in cell lines that formed highly vascularized tumors. A similar angiogenic-factor-expression pattern was found in the corresponding xenografts, with the exception of VEGF. In most cell lines, this factor had low expression in vitro which was strongly enhanced in vivo. Although all 8 melanoma cell lines were able to excise the angiostatin fragment from the plasminogen parent molecule in vitro, cell lines BLM and M14 showed the most potent angiostatin generation. In vitro angiostatin generation by cell lysates prepared from melanoma xenografts was comparable in all xenograft types. Thus, in our model system we found no correlation between angiostatin generation and vascular density. Our study has limited the number of pro-angiogenic factors that may be involved in melanoma angiogenesis, and provides evidence for the notion that regulation of tumor angiogenesis is dependent on multiple factors. Inhibition of angiogenesis for therapeutic purposes, therefore, should preferably not concentrate on a single factor. PMID- 10842190 TI - Expression of the ALK protein by anaplastic large-cell lymphomas correlates with high proliferative activity. AB - A variable fraction of anaplastic large-cell lymphomas (ALCLs) exhibits a t(2;5)(p23;q35) translocation that results in expression of the chimeric hyperphosphorylated protein NPM-ALK (p80). Tumor cells expressing NPM-ALK exhibit markedly enhanced proliferative activity, but comparative cellular kinetic studies on ALK(+) (ALK lymphomas) and ALK(-) lymphomas are lacking. The present study showed that ALK(+) lymphomas, detected with the monoclonal antibody ALKc (n = 17), had significantly higher average values for the proliferation-associated parameters mitotic index, ana/telophase index, growth index (x x mitotic index - apoptotic index, assuming x = 3), percentages of Ki-67(+) cells and fraction of cells expressing cyclin A or B or the cell cycle-regulatory protein p34(cdc2) than did ALK(-) ALCLs (n = 15). Whether this intense proliferative activity contributes to the good response to chemotherapy and favorable outcome of ALK(+) ALCLs remains to be assessed in a larger series of patients. Our findings support the notion that ALK(+) and ALK(-) ALCLs are 2 distinct disease entities. PMID- 10842191 TI - Disrupted transforming growth factor-beta signaling and deregulated growth in human biliary tract cancer cells. AB - Biliary tract carcinoma is a common neoplasm in Japan, and its treatment is difficult because it tends to promote fibrosis and easily invades surrounding tissues. To better characterize the biological features of this carcinoma, we investigated abnormalities in the transforming growth factor-beta (TGF-beta) signaling pathway in five human biliary tract cancer cell lines: RBE, KMBC, SK ChA-1, Mz-ChA-1, and Mz-ChA-2. We stably transfected into these cells the luciferase reporter plasmid carrying promoter of the plasminogen activator inhibitor-1 gene, the expression of which is stimulated by TGF-beta1. Treating the KMBC and Mz-ChA-1 cells with TGF-beta1 neither inhibited cell growth nor stimulated luciferase activity. In contrast, the RBE and Mz-ChA-2 cells responded well to TGF-beta1 treatment. TGF-beta1-treated SK-ChA-1 cells exhibited attenuated luciferase activity and their growth was not inhibited. Smad4 mRNA was not detected in SK-ChA-1 and Mz-ChA-1 cells by Northern blot analysis. Genetic analysis disclosed a nonsense mutation in the Mad homologue 2a domain of the Smad4 gene in the SK-ChA-1 cells and a heterozygous deletion in the TGF-beta type II receptor gene in the KMBC cells. Expression of the exogenous Smad4 gene in the Mz-ChA-1 cells by transient transfection restored their luciferase activity. When these TGF-beta1-insensitive and less-TGF-beta1-sensitive cell lines were xenografted into nude mice, they developed tumors that had more prominent, intervening fibrosis (desmoplasia) than the tumors caused by TGF-beta1-sensitive cells. Thus, a tight correlation between disruption of the TGF-beta signaling pathway and deregulated growth of cancer cells has been demonstrated in biliary tract carcinoma. This seems to be a critical event in this carcinoma and may also be correlated with stromal cell reaction in cancer invasion. PMID- 10842192 TI - Increased oxidative stress with gene alteration in urinary bladder urothelium after the Chernobyl accident. AB - We have previously shown that bladder urothelium of people living in the cesium 137 ((137)Cs)-contaminated areas of Ukraine demonstrates accumulation of stable p53 and p53 mutational inactivation, preferentially through G:C to A:T transition mutations at CpG dinucleotides, with a codon 245 hot spot. In the present study, we analyzed immuno-histochemically the relationship between oxidative stress markers and over-expression of p53 and H-ras in urinary bladder urothelium from 42 men with benign prostatic hyperplasia. Bladder mapping biopsies were obtained from 15 patients from a highly radiocontaminated area (group I), 14 patients from the less contaminated city of Kiev (group II) and 13 patients as a control group from "clean" (without radiocontamination) areas of Ukraine (group III). Irradiation cystitis with multiple foci of severe dysplasia and carcinoma in situ were observed in 15 of 15 (100%, group I) and 9 of 14 (64%, group II) cases, with 4 small transitional-cell carcinomas incidentally detected in groups I and II. Markedly elevated levels of inducible nitric oxide synthase (iNOS), cyclooxygenase 2 (COX-2) and 8-hydroxy-2;-deoxyguanosine (8-OHdG) were noted in these bladder urothelial lesions from groups I and II, accompanied by strong over expression of p53 and less H-ras expression. These findings support the hypothesis that iNOS, COX-2 and 8-OHdG in bladder urothelium are induced by long term exposure to low-dose radiation with a close relationship to p53 over expression that could predispose to bladder carcinogenesis. PMID- 10842193 TI - Molecular cloning and characterization of antigens expressed on rat tumor vascular endothelial cells. AB - We have previously prepared monoclonal antibodies (MAbs) against tumor vasculature using cultured rat tumor endothelial cells (TECs) isolated from solid KMT-17 tumors and identified 40 and 80 kDa antigens recognized by TES-17 and TES 23 MAbs, respectively. To clarify the nature of antigens on tumor vasculature, molecular cloning was conducted by screening the rat TEC cDNA library. Two antigens were detected: a 40 kDa OTS-8 antigen, which had been defined as a differentiation marker for osteoblastic lineage, and an 80 kDa CD44H antigen. Northern blots showed that OTS-8 mRNA was expressed exclusively in the lung, in addition to TECs, while CD44H mRNA was detected in the lung, intestine, spleen, thymus and peripheral blood cells, in addition to TECs. Immuno-histochemistry of KMT-17 tumors revealed that OTS-8 and CD44 were expressed on sprouting TECs. In addition, TES-23 MAb stained TECs of tubular vessels as well as sprouting TECs, but anti-rat CD44 MAbs stained only sprouting TECs. PCR showed that CD44 cDNA with a splice in exon 6 (CD44ex6) was present in rat TECs at low levels. Our results indicate that OTS-8 and CD44 are expressed on rat sprouting TECs and that TES-23 MAb recognizes CD44H on sprouting TECs as well as an unknown epitope on TECs of tubular vessels that could not be recognized by anti-rat CD44 MAbs. PMID- 10842194 TI - The tcf17 gene at chromosome 5q is not involved in the development of conventional renal cell carcinoma. AB - The human Kid-1 homolog Tcf17 has been cloned and assigned to chromosome 5q35.3. Since the chromosome 5q22-qter region is duplicated in approximately 50% of conventional renal-cell carcinomas, it was suggested that Tcf17 is involved in the development of renal tumors. We have analyzed Tcf17 mRNA in normal kidneys and genetically distinct types of renal-cell tumor and found it expressed in nearly all normal kidney and tumor samples. There was no correlation between allelic duplication and expression of Tcf17. We did not find mutations within the coding sequences but did detect deletions within the zinc finger domain in a small proportion of RNA molecules in both normal and tumor tissues. We found ubiquitous expression of human Tcf17 as well as rat Kid-1 in different types of human and rat tissue, indicating that the putative transcription-regulating activity of this zinc finger gene, in contrast to the published data, is not restricted to the kidney. The results of expression and mutation analyses suggest that the Tcf17 gene is not involved in the development of renal-cell tumors. PMID- 10842195 TI - Genetic complementation to non-tumorigenicity in cervical-carcinoma cells correlates with alterations in AP-1 composition. AB - The transcription factor AP-1 represents a central key element in the expression of human pathogenic papillomaviruses (HPV). We here propose a novel role for AP-1 as an essential component of an intracellular surveillance mechanism negatively controlling the proliferation of HPV-positive cells under in vivo conditions. The dissection of AP-1 composition in cervical-carcinoma cells revealed an inverse relationship between the Fos-related antigen Fra-1 and the tumorigenic phenotype. Cervical-carcinoma cell lines were either negative or expressed only low amounts of Fra-1 (jointly with c-Fos) within their AP-1 complexes. Somatic-cell hybridization technique was used to fuse different HPV-positive malignant cell lines. This resulted either in tumorigenic hybrids or in cells in which the malignant phenotype of the parental fusion partners was completely suppressed. The monitoring of AP-1 composition in electrophoretic mobility super-shift assays showed that the amount of Fra-1 was substantially increased within the AP-1 complex of non-malignant cells. In contrast, Fra-1 was even diminished in malignant hybrids, while c-Fos remained expressed. This correlation suggests that the concentration of Fra-1 within the AP-1 transcription complex might be an important marker for predicting the in vivo growth properties of HPV-positive cells. PMID- 10842196 TI - Threshold levels of gene expression of the melanoma antigen gp100 correlate with tumor cell recognition by cytotoxic T lymphocytes. AB - The level of expression of melanoma antigens (MA) may modulate the host immunologic response. Thus, the accurate measurement of MA expression may allow proper patient selection for antigen-specific therapies and yield important information for the evaluation of clinical results. In this study, we measured the absolute levels of MA messenger ribonucleic acid (mRNA) in tumor cell lines utilizing real-time quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). mRNA levels of MART-1, gp100, tyrosinase, TRP-1 and TRP-2 melanoma differentiation antigens and MAGE-1, MAGE-3 and ESO-1 cancer testis (CT) antigens were compared in 24 early-passage (<5 passages in culture) and 12 archival melanoma cell lines. MA mRNA expression was extremely variable among cell lines, occasionally reaching levels comparable to ribosomal RNA (rRNA). gp100 and MART-1 mRNA levels correlated with protein expression measurement obtained by FACS analysis. More significantly, a threshold of gp100 mRNA expression required for T cell stimulation and target-cell killing was identified. This threshold level corresponded to approximately 500 mRNA copies per 10(8) copies of rRNA. Our results suggest that the measurements of MA mRNA levels may yield useful information relevant to the interpretation of clinical outcome during antigen specific treatments. PMID- 10842197 TI - Flt3 ligand lessens the growth of tumors obtained after colon cancer cell injection in rats but does not restore tumor-suppressed dendritic cell function. AB - A defective function of the antigen-presenting cells may represent one of the ways used by cancer cells to escape the immune response. We have previously shown that human and rat colon carcinomas were infiltrated by dendritic cells that did not express the B7 co-stimulatory molecules required for inducing an efficient T cell response. Flt3 ligand is a cloned hematopoietic growth factor that markedly augments the number of functional dendritic and NK cells in lymphoid and non lymphoid tissues and exerts anti-tumor activity in various experimental models. We show here that repeated Flt3 ligand administration delays the s.c. growth of rat colon cancer cells in syngeneic animals without inducing tumor regression. In tumor-bearing animals, Flt3 ligand has a limited stimulatory effect on the antigen-presenting capacity of intra-tumoral and splenic dendritic cells, without restoring the high functional level of dendritic cells from tumor-free animals. Moreover, Flt3 ligand-mediated activation of NK cell cytotoxicity decreases when the tumor mass increases. Our results indicate that Flt3 ligand treatment of tumor-bearing animals does not sufficiently overcome tumor-induced immunosuppression to restore the inhibited functions of dendritic and NK cells and allow complete tumor regression. PMID- 10842198 TI - Monoclonal antibody 57B stains tumor tissues that express gene MAGE-A4. AB - Monoclonal antibody (Mab) 57B, which was raised against a recombinant MAGE-A3 protein, was tested for its ability to stain cells expressing various members of the MAGE-A gene family. COS-7 cells transfected with cDNAs encoding MAGE-A1, A2, A3, A4, A6, or A12 were stained, whereas those transfected with MAGE-A8, A9, A10, or A11 cDNAs were not. However, in tissue sections, we observed a different pattern of staining: the antibody effectively stained the tumors that expressed MAGE-A4 and only these tumors, regardless of the expression of the other MAGE-A genes. It seems, therefore, that at the level of MAGE gene expression found in tumors, a level clearly lower than that observed in transfected COS cells, only the MAGE-A4 protein can be reliably detected. We conclude that the 57B Mab should be useful for tumor diagnosis related to therapeutic vaccination involving MAGE A4. PMID- 10842199 TI - Restoration of functional defects in peripheral blood mononuclear cells isolated from cancer patients by thiol antioxidants alpha-lipoic acid and N-acetyl cysteine. AB - The ability of Alpha-Lipoic Acid (ALA) and N-Acetyl Cysteine (NAC), two active antioxidant agents, to correct in vitro the most significant functional defects of peripheral blood mononuclear cells (PBMC) isolated from advanced stage cancer patients was studied. The proliferative response of PBMC isolated from cancer patients to anti-CD3 monoclonal antibody (MAb) and the expression of CD25 (IL-2R) and CD95 (Fas) on unstimulated and anti-CD3 MAb-stimulated PBMC were studied, and the serum levels of proinflammatory cytokines IL-1, IL-6, TNFalpha as markers of pro-cachectic activity in cancer patients, and the serum levels of IL-2 and sIL 2R were assessed. Twenty patients (mean age 64.6 years) with cancer of lung, ovary, endometrium, and head and neck, all in advanced (III, IV) stage of disease, were studied. The serum levels of IL-1beta, IL-2, IL-6, TNFalpha, and sIL-2R were significantly higher in cancer patients than in normal subjects. The response of PBMC isolated from cancer patients to anti-CD3 MAb was significantly lower than that of controls. The addition of either ALA 0.001 mM or NAC 0.004 mM in the PBMC cultures stimulated with anti-CD3 MAb significantly increased the response of PBMC isolated from cancer patients and normal subjects. After 24 and 72 hr of culture with anti-CD3 MAb, the expression of CD25 and CD95 on PBMC isolated from cancer patients was significantly lower than that of PBMC isolated from normal subjects. The addition of either ALA or NAC into cultures of PBMC isolated from cancer patients significantly increased the percentage of cells expressing CD25 as well as those expressing CD95. The results of the present study show a favorable effect of antioxidant agents ALA and NAC on several important T-cell functions in vitro in advanced-stage cancer patients. PMID- 10842200 TI - Sensitisation of human carcinoma cells to the prodrug CB1954 by adenovirus vector mediated expression of E. coli nitroreductase. AB - The enzyme nitroreductase from E. coli can reduce the weak, monofunctional alkylating agent 5-(aziridin-1-yl)-2, 4-dinitrobenzamide (CB1954) to a potent cytotoxic species that generates interstrand crosslinks in DNA. Nitroreductase therefore has potential as a "suicide enzyme" for cancer gene therapy, as cells that express nitroreductase become selectively sensitive to the prodrug CB1954. We have incorporated a nitroreductase expression cassette into a replication defective adenovirus vector (Ad-CMV-ntr), which allowed efficient gene transfer to SK-OV-3 or IGROV-1 ovarian carcinoma cells. Nitroreductase levels increased in line with multiplicity of infection, and this was reflected in increasing sensitisation of the cells to CB1954, reaching an optimum (approx. 2, 000-fold sensitisation) with 25-50 p.f.u. per cell. Similar Ad-CMV-ntr-dependent sensitisation to CB1954 was seen in 3 of 6 low-passage primary ovarian tumour lines. Cells grown at low-serum concentration to inhibit proliferation remained equally susceptible to the Ad-CMV-ntr-dependent cytotoxicity of CB1954, indicating a distinct advantage over retroviral gene delivery and other popular enzyme-prodrug systems for human tumours with a low rate of cell proliferation. Additionally, cisplatin-resistant cells were sensitised towards CB1954 by Ad-CMV ntr as efficiently as the parental cells, indicating that the system could be effective in patients with cisplatin-resistant tumours. In a murine xenograft model for disseminated peritoneal carcinomatosis with ascites, treatment of nude mice bearing intraperitoneal SUIT2 tumours with Ad-CMV-ntr and CB1954 almost doubled the median survival from 14 to 26 days (p < 0.0001). PMID- 10842201 TI - Inhibition of progression to androgen-independence by combined adjuvant treatment with antisense BCL-XL and antisense Bcl-2 oligonucleotides plus taxol after castration in the Shionogi tumor model. AB - We have reported that antisense Bcl-2 oligodeoxynucleotide (ODN) delays progression to androgen independence in the androgen-dependent (AD) mouse Shionogi tumor model. Here, we characterize changes in bcl-xL, another important anti-apoptotic gene, and test the efficacy of adjuvant antisense Bcl-xL ODN therapy either alone or in combination with antisense Bcl-2 ODN and chemotherapy after castration in the Shionogi tumor model. Bcl-xL mRNA levels increased up to 3-fold postcastration and remained 1. 5-fold higher in androgen-independent (AI) recurrent tumors compared with AD tumors before castration. Treatment of Shionogi cells with antisense Bcl-xL ODN inhibited Bcl-xL expression in a dose-dependent and sequence-specific manner. Systemic administration of antisense Bcl-xL ODN in mice bearing Shionogi tumors after castration delayed emergence of AI recurrent tumors. We then examined whether combined adjuvant antisense Bcl-xL and/or Bcl-2 ODNs plus taxol (paclitaxel) therapy further delays time to AI progression. Combined treatment of Shionogi cells with antisense Bcl-xL and Bcl-2 ODNs significantly enhanced taxol chemosensitivity compared with either agent alone, reducing the IC(50) of taxol by more than 1 log. Apoptotic DNA laddering and cleavage of poly(ADP-ribose) polymerase were more substantial after treatment with combined antisense Bcl-2 and Bcl-xL ODNs plus taxol than that with either 2 agents. Adjuvant administration of antisense Bcl-xL and Bcl-2 ODNs plus micellar taxol resulted in a significantly delayed time to AI recurrence compared with administration of either 2 agents. Our findings suggest that Bcl-xL represents a suitable molecular target for antisense ODN strategy and illustrate the potential additive effects of multi-target pharmacology for cancer therapy. PMID- 10842202 TI - A transgenic mouse line that develops early-onset invasive gastric carcinoma provides a model for carcinoembryonic antigen-targeted tumor therapy. AB - In an attempt to obtain suitable in vivo models for optimizing new tumor therapy strategies for intestinal adenocarcinomas, carcinoembryonic antigen (CEA) promoter/SV40 T antigen gene constructs have been used to generate transgenic mice. One transgenic line (L5496), which contains a 424-bp CEA promoter/SV40 T antigen transgene, exclusively developed multi-focal carcinomas in the pyloric region of the stomach in 100% of the offspring. Tumors were already observable in 37-day-old animals as dysplastic cell foci within the mucosal layer. In 50-day old mice, the tumor mass was mainly restricted to the mucosa with invasive growth into the submucosal tissue. The animals became moribund at 100-130 days of age due to blockage of the pylorus. At this time, the tumor had penetrated into the duodenum and had invaded all tissue layers within the stomach. In contrast to most other stomach tumor models, this one perfectly matches the development of the most common stomach cancers found in humans. Furthermore, after crossing these mice with mice that are transgenic for the human CEA gene, the double transgenic offspring revealed expression of CEA in the resulting tumors. Thus, as well as being a model for studying gastric carcinoma development and prevention, this system should provide a useful preclinical model for CEA-targeted gastric tumor therapy. PMID- 10842203 TI - Anti-angiogenic treatment of human cancer: pitfalls and promises. PMID- 10842204 TI - Alcohol, smoking and papillomavirus infection as risk factors for esophageal squamous-cell papilloma and esophageal squamous-cell carcinoma in Italy. AB - Esophageal papilloma, an infrequent benign tumor, and esophageal squamous-cell carcinoma sometimes appear to be associated with human papillomavirus (HPV) infection, HPV being implicated in anogenital carcinogenesis. Our aim was to assess whether there is any epidemiological difference in terms of risk factors for papilloma and cancer. From 1989 to 1996, a total of 12,011 patients (53% male, median age 52.7 years) were submitted to esophagogastroduodenoscopy by our Digestive Endoscopy Service. The genome of HPV was sought by PCR using 2 different primer sets. Of the total, 42 subjects (0.35%), 50% male with a mean age of 45.1 years, were suffering from esophageal squamous-cell papilloma and 45 (0.37%), 91% male with a mean age of 63.0 years, from esophageal squamous-cell carcinoma. Of these patients, only 2 with papilloma were HPV(+). Compared with the general endoscopic population, patients with papilloma do not present significantly different characteristics (even in terms of frequency of esophagitis and hiatal hernia). Those with carcinoma differ significantly both from the general endoscopic population and from those with papilloma in that they are more often male (p < 0. 0001), older (p < 0.0001) and drinkers (p < 0.0001); they differ significantly only from the general population, but not from the papilloma patients, in smoking habits. Papilloma appears to be neither a lesion involving a risk of development into a malignancy nor a marker for any such risk. Environmental factors, such as alcohol and smoking, appear to play a decisive role in esophageal carcinogenesis in northern Italy. PMID- 10842205 TI - Sunscreens and cutaneous malignant melanoma: an Italian case-control study. AB - The possible relation between use of sunscreens and the risk of cutaneous malignant melanoma (CMM) was investigated in a case-control study conducted in 27 Italian centres on 542 incident, histologically confirmed cases and 538 controls admitted for acute, other than neoplastic or dermatologic conditions. Compared with subjects reporting never sunscreen use, the multivariate odds ratios (OR), after allowance for age, sex, geographic area, education, skin, eye and hair colour, freckles, number of naevi, history of sunburns, tanning pattern and duration of sunny vacations, were 0.97 (95% confidence interval [CI] 0.69 to 1.35) for those reporting "sometimes" and 0.80 (95% CI 0.54 to 1.17) for those reporting "often " sunscreen use. With reference to type of product most frequently used, the ORs were 0.96 (95% CI 0.52 to 1.77) for minimal, 0.90 (95% CI 0.63 to 1.28) for moderate and 1.41 (95% CI 0.85 to 2. 35) for high sunburn protection factor. With reference to duration of use, the OR was 0.86 (95% CI 0.58 to 1.29) for use started since >/=20 years. None of the corresponding trends in risks were significant. The ORs for sunscreen use were similar across strata of major identified covariates and, if anything, tended to decline after multivariate analysis. The present findings confirm that sunscreen use, as currently adopted in Italy, is not appreciably related to subsequent CMM risk. PMID- 10842206 TI - Early onset and familial predisposition to prostate cancer significantly enhance the probability for breast cancer in first degree relatives. AB - Genetic predisposition accounts for >/=10% of all cancer of the prostate (CaP) and is therefore considered a major risk factor, together with age and ethnic origin. Several epidemiological studies have suggested that familial clustering of CaP may be associated with an increased frequency of breast and other cancers among relatives. In order to correlate the incidence of CaP with prevalence of breast and other cancers, we have performed uni- and multi-variate analyses on 691 complete pedigrees including probands, who were consecutive patients with confirmed CaP treated in three French urological departments. We have shown a significantly higher risk (RR = 2.3, p = 0.01) to develop breast cancer in families with multiple than in those with a single CaP. Risk of observing other types of cancer within these families was not significant. We then calculated the breast cancer risk in early onset prostate cancer families, and observed a relative risk that is even more significant (RR = 5.5, p = 0.002). Furthermore, the risk was >30 times that a proband's mother have breast cancer if CaP occurred below 55 years of age, rather than after 75 years (p = 0.003). This study has therefore shown for the first time, the relatively high penetrance for breast cancer in relatives of early onset CaP patients. PMID- 10842207 TI - Osteoblast-derived TGFbeta-1 modulates matrix degrading protease expression and activity in prostate cancer cells PMID- 10842208 TI - Lesions in the budgerigar vocal control nucleus NLc affect production, but not memory, of english words and natural vocalizations. AB - This study investigates the role of the psittacid (parrot) central nucleus of the lateral neostriatum (NLc) in the production of learned English and natural vocalizations. Anatomic data have led researchers to define NLc alternately as the parrot homologue (Paton et al. [1981] J Neurosci. 11:1279-1288) or analogue (Striedter [1994] J Comp Neurol. 343:35-56) of the songbird high vocal center. Although numerous functional and electrophysiological studies have identified the role of various songbird vocal control nuclei, few similar functional studies have been performed in parrots, particularly with respect to NLc. In this study, both novel behavioral techniques and precise neurochemical lesions have been used to investigate the role of NLc in the production of learned vocalizations. The results suggest that NLc is involved in the production of, but not memory for, learned English and natural vocalizations. Specifically, NLc lesions disrupted the amplitude of amplitude-modulated vocalizations, but did not affect the frequency of the dominant or carrier signal of these vocalizations. These data provide some of the first evidence for the functional role of a parrot vocal control nucleus. PMID- 10842209 TI - Formation of cadherin-expressing brain nuclei in diencephalic alar plate divisions. AB - During the formation of brain nuclei, the vertebrate neural tube is partitioned into distinct embryonic divisions. In this study, the expression of three members of the cadherin family of adhesion molecules (cadherin-6B, cadherin-7, and R cadherin) was mapped to study the differentiation of gray matter in the divisions of the diencephalic alar plate of chicken embryos from embryonic day 3 (E3) to E10. At early stages of development (E3-E4), each cadherin is expressed in restricted regions of the diencephalic wall of the neural tube. The borders of some of the expression domains coincide with divisional boundaries. As the mantle layer is formed and increases in thickness from E4 to E8, morphologically discernible aggregates of cells appear that express the three cadherins differentially. These aggregates represent the anlagen of specific diencephalic brain nuclei, e.g., the lateroanterior nucleus, the ventral geniculate nucleus, the nucleus rotundus, the perirotundic area, the principal precommissural nucleus, and the lateral spiriform nucleus. Most of the cadherin-expressing diencephalic nuclei studied in this work apparently derive from a single embryonic division and remain there. The divisional boundaries are replaced gradually by the borders of cadherin-expressing brain nuclei. The current results support the idea that cadherins confer differential adhesiveness to developing structures of gray matter in the diencephalic alar plate. Moreover, they suggest that each cadherin plays a role in the formation of specific brain nuclei within the diencephalic divisions. PMID- 10842210 TI - Morphologic fate of diencephalic prosomeres and their subdivisions revealed by mapping cadherin expression. AB - The expression of four cadherins (cadherin-6B, cadherin-7, R-cadherin, and N cadherin) was mapped in the diencephalon of chicken embryos at 11 days and 15 days of incubation and was compared with Nissl stains and radial glial topology. Results showed that each cadherin is expressed in a restricted manner by a different set of embryonic divisions, brain nuclei, and their subregions. An analysis of the segmental organization based on the prosomeric model indicated that, in the mature diencephalon, each prosomere persists and forms a coherent domain of gray matter extending across the entire transverse dimension of the neural tube, from the ventricular surface to the pial surface. Moreover, the results suggest the presence of a novel set of secondary subdivisions for the dorsal thalamus (dorsal, intermediate, and ventral tiers and anteroventral subregion). They also confirm the presence of secondary subdivisions in the pretectum (commissural, juxtacommissural, and precommissural). At most of the borders between the prosomeres and their secondary subdivisions, changes in radial glial fiber density were observed. The diencephalic brain nuclei that derive from each of the subdivisions were determined. In addition, a number of previously less well-characterized gray matter regions of the diencephalon were defined in more detail based on the mapping of cadherin expression. The results demonstrate in detail how the divisions of the early embryonic diencephalon persist and transform into mature gray matter architecture during brain morphogenesis, and they support the hypothesis that cadherins play a role in this process by providing a framework of potentially adhesive specificities. PMID- 10842211 TI - Distribution of CGRP-like immunoreactivity in the chick and quail brain. AB - Calcitonin gene-related peptide (CGRP)-containing neurones have been implicated in the transmission of visceral sensory information to the cortex and in the control of arterial blood pressure in mammals. However, little is known about its function in other vertebrates. As a first step toward investigating the function of CGRP in birds, its distribution was studied in the domestic chick and quail brain by means of immunocytochemistry, by using antibodies against rat CGRP. The distribution of CGRP immunoreactivity in the chick and quail central nervous system was found to be similar. CGRP-immunoreactive (CGRPi) perikarya were not present in the telencephalon. In the diencephalon, CGRPi perikarya were present mainly in the shell of the thalamic nucleus ovoidalis, the nucleus semilunaris paraovoidalis, the nucleus dorsolateralis posterior thalami, and in the hypothalamic nucleus of the ansa lenticularis. In the brainstem, CGRPi perikarya were present in the nucleus mesencephalicus nervi trigemini, the nucleus tegmenti ventralis, the locus coeruleus, the nucleus linearis caudalis and in the parabrachial region. In addition CGRPi perikarya were found in the motor nuclei of the III, IV, V, VI, VII, IX, X, and XII cranial nerves. The telencephalon contained CGRPi fibres within the paleostriatal complex (mainly in the ventral paleostriatum), parts of the neostriatum and ventral hyperstriatum, parts of the archistriatum, and the septum. In the diencephalon, the densest plexus of CGRPi fibres was observed in the dorsal reticular thalamus. A less dense CGRPi innervation was present in some dorsal thalamic nuclei and in the medial and periventricular hypothalamus. The pretectum and midbrain tegmentum also contained CGRPi fibres, whereas the optic tectum was virtually devoid of immunolabelling. Scattered CGRPi fibres were observed in the central grey and neighbouring pontine areas. Some of the sensory fibres of the trigeminal, vagal, glossopharyngeal, and spinal nerves were also CGRPi. The results of comparative studies indicate that the presence of CGRP in some thalamo-telencephalic projections is a primitive feature of the forebrain of amniotes. Therefore, the brain areas giving rise to and receiving such a projection in different vertebrates, are likely to be homologous. PMID- 10842212 TI - SNAP-25 regulation during adrenal gland development: comparison with differentiation markers and other SNAREs. AB - Synaptosomal-associated protein of 25 kDa (SNAP-25) is one of a limited number of soluble N-ethylmaleimide-sensitive fusion attachment protein receptors (SNAREs) that play a major role in membrane docking of synaptic vesicles and secretory granules during regulated exocytosis. We have previously shown that SNAP-25 levels differ between noradrenergic and adrenergic chromaffin cell populations of the adult adrenal gland. We examine SNAP-25 expression by immunofluoresence in cells of the sympathoadrenal lineage in the rat during late embryonic and postnatal development. In parallel, tyrosine hydroxylase was used to identify sympathoadrenal cells, phenylethanolamine N-methyltransferase to distinguish adrenergic from noradrenergic chromaffin cells, and chromogranin A to define the presence of secretory granules. In addition, SNAP-25 protein and mRNA levels were followed in adrenal gland extracts by immunoblotting and reverse transcription polymerase chain reaction (RT-PCR). Protein levels were compared with those of other molecules also implicated in organelle trafficking, including syntaxin 1 and vesicle-associated membrane protein (VAMP-2) and the nonneuronal analogues SNAP-23 and cellubrevin. This study provides evidence that SNAP-25 is expressed early during development in sympathoadrenal neurons and migrating cells. It is detected in intra-adrenal chromoblasts as soon as they enter the adrenal primordium. Its differential expression between catecholamine chromaffin cell phenotypes is already evident from the 17th embryonic day, future noradrenergic cells appearing to express higher levels than adrenergic cells. The granule maturation marker chromogranin A is expressed in chromaffin cells later than SNAP 25. Both SNAP-25 protein and mRNA increased rapidly in the adrenal gland in the perinatal period to peak during the first postnatal week, after which levels dropped dramatically to adult values. In contrast, levels of both syntaxin and SNAP-23 appeared to remain fairly constant throughout adrenal gland development. VAMP-2 expression increased gradually around birth to reach maximal levels during the first two postnatal weeks, and then decreased slightly. Cellubrevin levels also appeared to increase gradually until adult values were attained by the end of the second postnatal week. The threefold increase of SNAP-25 mRNA shortly after birth compared to the low adult levels suggests that during this period SNAP-25 is implicated in additional functions than regulated secretion, possibly associated with cellular growth or maturation. PMID- 10842213 TI - Immunohistochemical localization of prostaglandin EP3 receptor in the rat nervous system. AB - The prostaglandin EP3 receptor (EP3R) subtype is believed to mediate large portions of diverse physiologic actions of prostaglandin E2 in the nervous system. However, the distribution of EP3R protein has not yet been unveiled in the peripheral or central nervous systems. The authors raised a polyclonal antibody against an amino-terminal portion of rat EP3R that recognized specifically the receptor protein. In this study, immunoblotting analysis with this antibody showed several immunoreactive bands with different molecular weights in rat brain extracts and in membrane fractions of recombinant EP3R expressing culture cells, and treatment with N-glycosidase shifted those immunoreactive bands to an apparently single band with a lower molecular weight, suggesting that EP3R proteins are modified posttranslationally with carbohydrate moieties of various sizes. The authors performed immunohistochemical investigation of EP3R in the rat brain, spinal cord, and peripheral ganglia by using the antibody. EP3R-like immunoreactivity was observed in many and discrete regions of the rostrocaudal axis of the nervous system. The signals were particularly strong in the anterior, intralaminar, and midline thalamic nuclear groups; the median preoptic nucleus; the medial mammillary nucleus; the superior colliculus; the periaqueductal gray; the lateral parabrachial nucleus; the nucleus of the solitary tract; and laminae I and II of the medullary and spinal dorsal horns. Sensory ganglia, such as the trigeminal, dorsal root, and nodose ganglia, contained many immunopositive neurons. Neuronal cells in the locus coeruleus and raphe nuclei exhibited EP3R-like immunoreactivity. This suggests that EP3R plays regulatory roles in the noradrenergic and serotonergic monoamine systems. Autonomic preganglionic nuclei, such as the dorsal motor nucleus of the vagus nerve, the spinal intermediolateral nucleus, and the sacral parasympathetic nucleus, also contained neuronal cell bodies with the immunoreactivity, implying modulatory functions of EP3R in the central autonomic nervous system. The characteristic distribution of EP3R provides valuable information on the mechanisms for various physiologic actions of prostaglandin E2 in the central and peripheral nervous systems. PMID- 10842214 TI - Distribution and innervation of lateral line organs in the channel catfish. AB - The lateral line system of the channel catfish is formed by mechanoreceptive neuromasts located within five pairs of cephalic and one pair of trunk canals, as well as superficial lines of neuromasts, termed accessory and/or pit lines. Five pairs of pit lines occur on the head, and three pairs of superficial lines occur on the trunk. In addition to these mechanoreceptors, which are found in most teleost fishes, catfish also possess a total of over 4000 electroreceptive ampullary organs scattered over the entire body. The lateral line receptors are innervated by five pairs of lateral line nerves whose rami are secondarily associated with facial and trigeminal fibers that innervate taste buds and the dermis of the skin, respectively. The neuromasts of the trunk canal and the ramules of the posterior lateral line nerve that innervate them seem to be organized in a segmental pattern. The same is true for the intervertebral ramules of the recurrent facial ramus, which innervate the external taste buds on the trunk. The fibers of the gustatory and lateral line systems may use the neural crest, the developing spinal nerves, or both, to establish this segmental pattern. In this context, it may not be surprising that there is an intimate relationship among each of the sensory systems in the trunk. PMID- 10842215 TI - Postembryonic development of the dorsal longitudinal flight muscle and its innervation in Manduca sexta. AB - The neuromuscular systems of holometabolous insects must be remodeled during metamorphosis to allow striking behavioral changes, such as the acquisition of flight. The fast contracting dorsal longitudinal flight muscle (DLM) of Manduca arises from an anlage containing both remnants of specific larval dorsal body wall muscles and extrinsic myoblasts. In the mesothorax, the DLM is innervated by five persisting larval motoneurons: one in the mesothoracic and four in the prothoracic ganglion. These motoneurons innervate two slowly contracting body wall muscles in the larva. 2 days before pupation, the DLM template fibers begin to degenerate, whereas other muscles remain intact until pupation. Correspondingly, the motor terminals retract from the template fibers while they remain on other muscle fibers until pupation. Accumulation and proliferation of putative myoblasts also starts 2 days before pupation in close spatial relationship to the retracted motor tufts around the degenerating larval template fibers. Proliferation increases through the early pupal stages, and is detected within the anlage until the ninth day after pupation. 2 days after pupation, the anlage splits into five bundles, each innervated by one motoneuron. Striations occur on the seventh day after pupation when the growing motor axons reach the attachment sites. Subsequently, the muscle grows in volume and higher-order motor branches are formed. Within the central nervous system, there is dramatic regression of larval dendrites followed by growth of new dendrites as the persistent motoneurons assume their new role in flight behavior. Both central and peripheral remodeling follow similar time courses. PMID- 10842216 TI - Activity-dependent reconfiguration of the effective dendritic field of motoneurons. AB - A neuron in vivo receives a continuous bombardment of synaptic inputs that modify the integrative properties of dendritic arborizations by changing the specific membrane resistance (R(m)). To address the mechanisms by which the synaptic background activity transforms the charge transfer effectiveness (T(x)) of a dendritic arborization, the authors simulated a neuron at rest and a highly excited neuron. After in vivo identification of the motoneurons recorded and stained intracellularly, the motoneuron arborizations were reconstructed at high spatial resolution. The neuronal model was constrained by the geometric data describing the numerized arborization. The electrotonic structure and T(x) were computed under different R(m) values to mimic a highly excited neuron (1 kOhm x cm(2)) and a neuron at rest (100 kOhm x cm(2)). The authors found that the shape and the size of the effective dendritic fields varied in the function of R(m). In the highly excited neuron, the effective dendritic field was reduced spatially by switching off most of the distal dendritic branches, which were disconnected functionally from the somata. At rest, the entire dendritic field was highly efficient in transferring current to the somata, but there was a lack of spatial discrimination. Because the large motoneurons are more sensitive to variations in the upper range of R(m), they switch off their distal dendrites before the small motoneurons. Thus, the same anatomic structure that shrinks or expands according to the background synaptic activity can select the types of its synaptic inputs. The results of this study demonstrate that these reconfigurations of the effective dendritic field of the motoneurons are activity-dependent and geometry dependent. PMID- 10842217 TI - Orbitofrontal sulci of the human and macaque monkey brain. AB - The present study investigated the orbitofrontal sulci in 100 normal adult human cerebral hemispheres by using magnetic resonance images that were transformed into the standardized proportional stereotaxic space most commonly used, that of Talairach and Tournoux (Talairach and Tournoux [1988]. Co-planar stereotaxic atlas of the human brain. New York: Thieme). The patterns formed by the individual sulci were then examined and compared with those of the less convoluted macaque monkey brain. Four sulci forming a similar sulcal pattern were identified in both species. The olfactory sulcus occupies the most medial position forming the lateral border of the gyrus rectus. Lateral to this, the medial, lateral, and transverse orbital sulci form a pattern often resembling an "H," "X," or "K." These sulci divide the orbitofrontal cortex into four major gyri: the medial, lateral, anterior, and posterior orbital gyri. Three major types of sulcal pattern were identified in both species based on the arrangement of these orbital sulci. Additional sulci were observed in the human brain, creating more complex patterns. Probability maps were constructed for the four main orbitofrontal sulci of the human brain. These maps provide a statistical description of the variability of the location of the orbitofrontal sulci within the three-dimensional coordinate system of Talairach and Tournoux (Talairach and Tournoux [1988]. Co-planar stereotaxic atlas of the human brain. New York: Thieme). Because these maps may be directly compared with any image transformed into the same standardized space, they provide a valuable tool for identifying and describing the location of functional or structural changes in the orbitofrontal region of the human brain. PMID- 10842218 TI - Unipolar brush cell axons form a large system of intrinsic mossy fibers in the postnatal vestibulocerebellum. AB - The unipolar brush cells (UBCs), a class of neurons recently identified in the granular layer of the vestibulocerebellum, receive excitatory synaptic input from mossy fibers (MFs) in the form of a giant glutamatergic synapse. UBCs are provided with axons that bear synaptic endings situated at the center of glomeruli, similar to cerebellar MF afferents. A single MF stimulus evokes a prolonged train of action potentials in the UBC (Rossi et al., 1995), which is presumably distributed to postsynaptic targets. Knowledge of the synaptic connections of UBC axons is essential to define the role of these cells in the integration of vestibular signals in the cerebellar circuitry. To evaluate these connections, the nodulus (folium X) was isolated from vermal slices of postnatal day 8 mice, cultured for 2-4 or 15-30 days in vitro, and studied by electron and fluorescence microscopy. The peak of degeneration of extrinsic MF terminals, which have been severed from the parent cell bodies, was observed at 2 days in vitro (DIV). Quantification of degenerating and nondegenerating (e.g., intrinsic) MF terminals indicated that about half of the MF terminals were provided by local UBC axons synapsing on dendrites of granule cells and other UBCs. The proportion of nondegenerating vs. degenerating MF terminals terminating on UBCs also indicated that approximately two-thirds of the intrinsic MFs are involved in UBC UBC connections. In long-term cultures, the granular layer appeared well preserved and the UBC axons formed an extensive system of MF collaterals. It is suggested that UBCs may act by spatially amplifying vestibular inputs carried by extrinsic MFs. PMID- 10842219 TI - Neuroanatomy of cells expressing clock genes in Drosophila: transgenic manipulation of the period and timeless genes to mark the perikarya of circadian pacemaker neurons and their projections. AB - Subsets of brain neurons expressing the clock genes period (per) and timeless (tim) are involved in the generation of circadian behavioral rhythms. However, current knowledge of projection patterns of these neurons is limited to those immunoreactive to an antibody against a crustacean neuropeptide. The GAL4 expression system was utilized to visualize neuronal processes from all per and tim-expressing neurons in the central nervous system. Each of two types of GAL4 driver fusion genes, per-gal4 or tim-gal4, was combined in transgenic flies with marker genes-lacZ, and sequences encoding green fluorescent protein or TAU protein-under the control of the GAL4-responsive element UAS. This allowed visualization of the cytoplasm of GAL4-expressing cells. Thus, neurites of clock neurons in the adult brain as well as those of larvae and pupae were revealed. Among the anatomical patterns revealed by per-gal4- or tim-gal4-driven marker expression were a previously unknown, dorsally located neuronal cluster, along with the projections of these cells and of other dorsal neurons characterized in earlier studies only by the location of their perikarya. The similarity of projections from PER- or TIM-containing neurons during development to those in the adult implies that these features of mature clock neurons are established by the larval stages. Neurons that have never been identified as PER- or TIM immunoreactive were also visualized in this assay system, indicating promoter activity of the clock genes in these cells and suggesting that their products cannot accumulate to detectable levels in certain neurons. PMID- 10842220 TI - Olivocochlear neurons sending axon collaterals into the ventral cochlear nucleus of the rat. AB - The olivocochlear projection constitutes the last stage of the descending auditory system in the mammalian brain. Its neurons reside in the superior olivary complex (SOC) and project to the inner and outer hair cell receptors in the cochlea. Olivocochlear neurons were also reported to send axon collaterals into the cochlear nucleus, but controversies about their number and about species differences persist. By injecting the fluorescent retrograde axonal tracers diamidino yellow and fast blue into the cochlea and the ventral cochlear nucleus (VCN), we studied the distribution and number of olivocochlear neurons with and without axon collaterals into the VCN of the rat. We found that olivocochlear neurons residing in the lateral superior olive (LSO), the intrinsic lateral olivocochlear cells (intrinsic LOCs), do not send axon collaterals into the VCN. By contrast, a majority, and possibly all, olivocochlear neurons residing in the ventral nucleus of the trapezoid body (VNTB), the medial olivocochlear cells (MOCs), do have such axon collaterals. These cells may thus affect processing in the ascending auditory pathway at the level of the receptors and concurrently at the level of the secondary sensory neurons in the cochlear nucleus. Belonging to the lateral olivocochlear system, shell neurons reside around the LSO and form a third group of olivocochlear cells (shell LOCs). Like intrinsic LOCs, they innervate the inner hair cells, but like MOCs they do, by means of axon collaterals, project into the VCN. These findings have implications for understanding both auditory signal processing and the plasticity responses that occur following loss of cochlear function. PMID- 10842221 TI - Ascending spinal systems in the fish, Prionotus carolinus. AB - The fin rays of the pectoral fin of the sea robins (teleostei) are specialized chemosensory organs heavily invested with solitary chemoreceptor cells innervated only by spinal nerves. The rostral spinal cord of these animals is marked by accessory spinal lobes which are unique enlargements of the dorsal horn of the rostral spinal segments receiving input from the fin ray nerves. Horseradish peroxidase (HRP) and 1,1;-dioctadecyl-3,3,3', 3'-tetramethylindocarbocyanine perchlorate (diI) were used as anterograde and retrograde tracers to examine the connectivity of these accessory lobes and the associated ascending spinal systems in the sea robin, Prionotus carolinus. The majority of dorsal root fibers terminate within the accessory lobes at or nearby their level of entrance into the spinal cord. A few dorsal root axons turn rostrally in the dorsolateral fasciculus to terminate in the lateral funicular complex situated at the spinomedullary junction. The lateral funicular complex also receives a heavy projection from the ipsilateral accessory lobes. In addition, it contains a few large neurons that project back onto the accessory lobes. Injections of either diI or HRP into the lateral funicular complex label fibers of the medial lemniscus which crosses the midline in the caudal medulla to ascend along the ventral margin of the contralateral rhombencephalon. Within the medulla, fibers leave the medial lemniscus to terminate in the inferior olive and in the ventrolateral medullary reticular formation. Upon reaching the midbrain, the medial lemniscus turns dorsally to terminate heavily in a lateral division of the torus semicircularis, in the ventral optic tectum, and in the lateral subnucleus of the nuc. preglomerulosus of the thalamus. Lesser projections also reach the posterior periventricular portion of the posterior tubercle with a few fibers terminating along the ventral, posterior margin of the ventromedial (VM) nucleus of the thalamus. The restricted projection to the ventral tectum is noteworthy in that this part of the tectum maintains the representation of the ventral visual field, that is, the area in which the fin rays lie. A prominent spinocerebellar system is also evident. Both direct and indirect spinocerebellar fibers can be followed through the dorsolateral fasciculus, with or without relay in the lateral funicular nucleus and terminating in a restricted portion of the granule cell layer of the ipsilateral corpus cerebelli. The similarities in connectivity of the spinal cord between the sea robins and other vertebrates are striking. It is especially notable because sea robins utilize the chemosensory input from the fin rays to localize food in the environment. Thus, although these fish use their spinal chemosense as other fishes use their external taste systems, the spinal chemosense apparently relies on the medial lemniscal system to guide this chemically driven feeding behavior. PMID- 10842222 TI - Distribution of voltage-gated sodium channel alpha-subunit and beta-subunit mRNAs in human hippocampal formation, cortex, and cerebellum. AB - The distribution of mRNAs encoding voltage-gated sodium channel alpha subunits (I, II, III, and VI) and beta subunits (beta1 and beta2) was studied in selected regions of the human brain by Northern blot and in situ hybridisation experiments. Northern blot analysis showed that all regions studied exhibited heterogenous expression of sodium channel transcripts. In situ hybridisation experiments confirmed these findings and revealed a predominantly neuronal distribution. In the parahippocampal gyrus, subtypes II and VI and the beta subunit mRNAs exhibited robust expression in the granule cells of the dentate gyrus and pyramidal cell layer of the hippocampus. Subtypes I and III showed moderate expression in granule cells and low expression in the pyramidal cell layer. Distinct expression patterns were also observed in the cortical layers of the middle frontal gyrus and in the entorhinal cortex. In particular, all subtypes exhibited higher levels of expression in cortical layers III, V, and VI compared with layers I and II. All subtypes were expressed in the granular layer of the cerebellum, whereas specific expression of subtypes I, VI, beta1, and beta2 mRNAs was observed in Purkinje cells. Subtypes I, VI, and beta1 mRNAs were expressed, at varying levels, in the pyramidal cells of the deep cerebellar nuclei. These data indicate that, as in rat, human brain sodium channel mRNAs have a distinct regional distribution, with individual cell types expressing different compliments of sodium channels. The differential distribution of sodium channel subtypes suggest that they have distinct roles that are likely to be of paramount importance in maintaining the functional heterogeneity of central nervous system neurons. PMID- 10842223 TI - Development of inhibitory circuitry in visual and auditory cortex of postnatal ferrets: immunocytochemical localization of calbindin- and parvalbumin-containing neurons. AB - The inhibitory neurotransmitter gamma-aminobutyric acid (GABA) is thought to play an important role in activity-dependent stages of brain development. Previous studies have shown that different functional subclasses of cortical GABA containing neurons can be distinguished by antibodies to the calcium-binding proteins parvalbumin and calbindin. Thus insight into the development of distinct subsets of inhibitory cortical circuits can be gained by studying the development of these calcium-binding protein-containing neurons. Previous studies in several mammalian species have suggested that calcium-binding proteins are upregulated in sensory cortex when thalamocortical afferents arrive. In ferrets, the ingrowth of thalamic axons into cortex occurs well into postnatal development, allowing access to early stages of cortical development and calcium-binding protein expression. We find in ferrets that both parvalbumin- and calbindin immunoreactivity are present in primary visual and primary auditory cortex long before thalamocortical synapse formation, but that there is a sharp decline in immunoreactivity by postnatal day 20. Day 20 in ferrets corresponds to postnatal day 1 in cats, and thus previous studies in postnatal cats would have missed this early pattern of calcium-binding protein distribution. Another surprising finding is that the proportion of parvalbumin- and calbindin-immunoreactive neurons peaks secondarily late in development, between P60 and adulthood. This result suggests that the parvalbumin- and calbindin-containing subclasses of nonpyramidal neurons remain immature until late in the critical period for cortical plasticity, and that they are positioned to play an important role in experience-dependent modification of cortical circuits. PMID- 10842224 TI - Localisation of cannabinoid receptors in the rat brain using antibodies to the intracellular C-terminal tail of CB. AB - The CB(1)-type cannabinoid receptor mediates physiologic effects of Delta(9) tetrahydrocannabinol, the psychoactive ingredient of the drug marijuana. In this report, the authors analyse the expression of CB(1) in the rat brain by using antibodies to the C-terminal 13 amino acids of the receptor. Western blot analysis of rat brain membranes revealed a prominent immunoreactive band with a molecular mass ( approximately 53 kDa) consistent with that predicted for CB(1) from the rat cDNA sequence. In addition, however, less intense immunoreactive bands corresponding to glycosylated ( approximately 62 kDa) and putative N terminally shorter ( approximately 45 kDa) isoforms of CB(1) were detected. The distribution of CB(1)-immunoreactivity in rat brain was similar to the distribution of binding sites for radiolabelled cannabinoids, with high levels of expression in the olfactory system, the hippocampal formation, the basal ganglia, the cerebellum, and the neocortex. This provides important evidence that CB(1) is likely to be largely responsible for mediating effects of cannabinoids in the brain. CB(1) immunoreactivity was associated with nerve fibre systems and axon terminals but was not detected in neuronal somata. This is consistent with the presynaptic inhibitory effects of cannabinoids on neurotransmitter release in the brain. Detailed immunocytochemical analysis of anatomically or functionally related regions of the brain revealed the location of CB(1) receptors within identified neural circuits. Determination of the cellular and subcellular location of CB(1) within known neuronal circuits of the brain provides an anatomic framework for interpretation of the neurophysiologic and behavioural effects of cannabinoids. PMID- 10842225 TI - Delayed loss of small dorsal root ganglion cells after transection of the rat sciatic nerve. AB - The present study deals with changes in numbers and sizes of primary afferent neurons (dorsal root ganglion [DRG] cells) after sciatic nerve transection. We find that this lesion in adult rats leads to death of some DRG cells by 8 weeks and 37% by 32 weeks after the lesion. The loss of cells appears earlier in and is more severe in B-cells (small, dark cells with unmyelinated axons) than A-cells (large, light cells with myelinated axons). With regard to mean cell volumes, there is a tendency for both categories of DRG cells to be smaller, but except for isolated time points, these differences are not statistically significant. These findings differ from most earlier reports in that the cell loss takes place later than usually reported, that the loss is more severe for B-cells, and that neither A- or B-cells change size significantly. Accordingly, we conclude that sciatic nerve transection in adult rats leads to a slowly developing but relatively profound loss of primary afferent neurons that is more severe for B cells. These results can serve as a basis for studies to determine the effectiveness of trophic or survival factors in avoiding axotomy induced cell death. PMID- 10842226 TI - mu-opioid receptors are present in vagal afferents and their dendritic targets in the medial nucleus tractus solitarius. AB - Ligands of the mu-opiate receptor (MOR) are known to influence many functions that involve vagal afferent input to the nucleus tractus solitarius (NTS), including cardiopulmonary responses, gastrointestinal activity, and cortical arousal. The current study sought to determine whether a cellular substrate exists for direct modulation of vagal afferents and/or their neuronal targets in the NTS by ligands of the MOR. Anterograde tracing of vagal afferents arising from the nodose ganglion was achieved with biotinylated dextran amine (BDA), and the MOR was detected by using antipeptide MOR antiserum. The medial subdivision of the intermediate NTS was examined by electron microscopy for the presence of peroxidase-labeled, BDA-containing vagal afferents and immunogold MOR labeling. MOR was present in both presynaptic axon terminals and at postsynaptic sites, primarily dendrites. In dendrites, MOR immunogold particles usually were located along extrasynaptic portions of the plasma membrane. Of 173 observed BDA-labeled vagal afferent axon terminals, 33% contained immunogold labeling for MOR within the axon terminal. Many of these BDA-labeled terminals formed asymmetric, excitatory-type synapses with dendrites, some of which contained MOR immunogold labeling. MORs were present in 19% of the dendrites contacted by BDA-labeled terminals but were present rarely in both the vagal afferent and its dendritic target. Together, these results suggest that MOR ligands modulate either the presynaptic release from or the postsynaptic responses to largely separate populations of vagal afferents in the intermediate NTS. These results provide a cellular substrate for direct actions of MOR ligands on primary visceral afferents and their second-order neuronal targets in NTS. PMID- 10842228 TI - Divergent backward projections from the anterior part of the inferotemporal cortex (area TE) in the macaque. AB - The organization of backward projections from the anterior part of the inferotemporal cortex (area TE) to the posterior part of the inferotemporal cortex (area TEO) was studied in the macaque monkey by using the anterograde tracer Phaseolus vulgaris-leucoagglutinin (PHA-L). The objectives of the study were to investigate this backward projection and to compare it with 1) the backward projections that have been described previously in the early sensory areas and 2) the forward projection from area TEO to area TE. After a single iontophoretic injection of PHA-L into area TE in three monkeys, a dense distribution of labeled terminals was observed in area TEO and in the ventral bank of the superior temporal sulcus (area PITd) that adjoined area TEO. A less dense distribution was observed in areas V4, V2, and V1. Clusters of labeled terminals in areas TEO and PITd extended more than 4 mm along the cortical surface. The forward projections from area TEO to area TE also were studied for comparison by reanalyzing two previous cases (Saleem et al. ?1993 Cerebral Cortex 3:454-464). These projections (from area TEO to area TE) were more focal than the terminations that occurred in area TEO after injections into area TE. Nine single axons projecting from area TE to areas TEO/PITd were reconstructed through serial sections. These showed variable, complex branching patterns with multiple arbors (1-12). Arbors were localized in layers 1-3 for four axons, in layer 1 for one axon, layers 5 and 6 for two axons, and in both layers 1-3 and layers 5-6 for two axons. Axons with horizontally elongated arbors confined to layer 1 were not predominant. The size of the individual arbors of these axons along their long axes tended to be larger (1.56 +/- 1.24 mm) than those of TEO-to-TE forward axons (<0.6 mm). Thus, the authors conclude that, like other backward systems described to date, those from area TE to areas TEO/PITd are divergent. However, single axons have more variable laminar patterns of terminal distribution than those in the other backward systems. PMID- 10842227 TI - Expression of nitric oxide synthase and soluble guanylyl cyclase in the developing olfactory system of Manduca sexta. AB - The gaseous messenger nitric oxide (NO), with its ability to mediate both intercellular and intracellular communication, can play important roles in mediating cellular communication in both the development and the function of the nervous system. The authors investigated the possible role of NO signaling in the developing olfactory system (antennal lobe) of the moth Manduca sexta. NO synthase (NOS), the enzyme that generates NO, was localized by using immunocytochemistry, in situ hybridization, and nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) histochemistry. Although NADPH-d staining appears to be a poor indicator of the presence of NOS in this system, immunocytochemistry and in situ hybridization reveal that NOS is expressed in the axons of olfactory receptor neurons throughout development and in the perineurial sheath that covers the brain early in development. NOS is present in axon terminals as they form protoglomeruli, raising the possibility that NO mediates cell-cell interactions during antennal lobe development. NO-sensitive soluble guanylyl cyclase (sGC), one of the best characterized targets of NO, was localized in the developing olfactory system by using in situ hybridization and immunocytochemistry for the Manduca sexta sGCalpha1 subunit. The ability of the developing olfactory system to respond to exogenous NO also was examined by using cyclic guanosine monophosphate immunocytochemistry. sGC is expressed in mechanosensory neurons in the developing antenna and in many antennal lobe neurons in both the medial and lateral cell body clusters. Thus, NOS and sGC are expressed in a pattern that suggests that this signaling pathway may mediate intercellular communication during development of the olfactory system in Manduca sexta. PMID- 10842229 TI - Formation of antennal lobe and mushroom body neuropils during metamorphosis in the honeybee, apis mellifera. AB - The projections to the mushroom bodies (mbs) have been clearly described in the brain of adult honeybees (Apis mellifera). Olfactory projection neurons arborize in the lip of the calyceal neuropil, whereas visual projection neurons project to the collar. To study the maturation of this pattern of innervation, as well as the development of uniglomerular projection neurons within the antennal lobes (als), we conducted the following three studies focused on the first four stages of pupal development: mass staining of olfactory projection neurons, single cell labeling of olfactory projection neurons, and simultaneous labeling of olfactory projection neurons and visual projection neurons. Examination of whole-mount preparations with the confocal laser scanning microscope revealed that the olfactory projection neurons achieved their adult arborization pattern within their main output region, the lip of the mb calyces, earlier during development (pupal stage 1) than their dendritic processes within their main input region, the al (pupal stage 2). Simultaneous labeling experiments showed further that the fiber terminals of olfactory projection neurons and visual projection neurons did not overlap but instead occupied their respective projection areas within the mb calyces as early as pupal stage 1. We conclude that selective innervation of different subregions of the calycal neuropil precedes the segregation of glomerular units within the antennal lobe neuropil, and that the Kenyon cells themselves provide a template for the innervation of olfactory and visual projection neurons. PMID- 10842230 TI - Rat somatosensory cerebropontocerebellar pathways: spatial relationships of the somatotopic map of the primary somatosensory cortex are preserved in a three dimensional clustered pontine map. AB - In the primary somatosensory cortex (SI), the body surface is mapped in a relatively continuous fashion, with adjacent body regions represented in adjacent cortical domains. In contrast, somatosensory maps found in regions of the cerebellar hemispheres, which are influenced by the SI through a monosynaptic link in the pontine nuclei, are discontinuous ("fractured") in organization. To elucidate this map transformation, the authors studied the organization of the first link in the SI-cerebellar pathway, the SI-pontine projection. After injecting anterograde axonal tracers into electrophysiologically defined parts of the SI, three-dimensional reconstruction and computer-graphic visualization techniques were used to analyze the spatial distribution of labeled fibers. Several target regions in the pontine nuclei were identified for each major body representation. The labeled axons formed sharply delineated clusters that were distributed in an inside-out, shell-like fashion. Upper lip and other perioral representations were located in a central core, whereas extremity and trunk representations were found more externally. The multiple clusters suggest that the pontine nuclei contain several representations of the SI map. Within each representation, the spatial relationships of the SI map are largely preserved. This corticopontine projection pattern is compatible with recently proposed principles for the establishment of subcortical topographic patterns during development. The largely preserved spatial relationships in the pontine somatotopic map also suggest that the transformation from an organized topography in SI to a fractured map in the cerebellum takes place primarily in the mossy fiber pontocerebellar projection. PMID- 10842231 TI - Baratin, a nonamidated neurostimulating neuropeptide, isolated from cockroach brain: distribution and actions in the cockroach and locust nervous systems. AB - During the purification of tachykinin-related peptides from the brain of the cockroach Leucophaea maderae, a few other peptides were collected in adjacent high-performance liquid chromatography fractions. Edman degradation, mass spectrometry, and chemical synthesis revealed that one of these peptides had the sequence DNSQWGGFA. This nonamidated nonapeptide was designated baratin and appears not to be related to any known insect peptide. Baratin was not found to be bioactive in the L. maderae hindgut or oviduct muscle contraction assay. (Both synthetic nonamidated and amidated baratin were tested.) To screen for possible sites of action, we raised a rabbit antiserum to baratin. We found baratin immunoreactive (BAR-IR) interneurons throughout the cockroach central nervous system. Some prominent brain neuropils were supplied by BAR-IR neuron processes: the central body, the calyx, and lobes of the mushroom bodies, parts of the optic lobe, and the tritocerebral neuropil. Additionally we found BAR-IR neurosecretory cells in the median neurosecretory cell group with processes supplying the storage lobe of the corpora cardiaca. In each of the thoracic and abdominal ganglia processes of BAR-IR projection neurons and local neurons were seen. The baratin antiserum also labeled neurons in the brain of the locust Locusta migratoria, some of which are similar to those of the cockroach. A prominent system of interganglionic BAR-IR processes was found in the locust subesophageal, thoracic, and abdominal ganglia. This was formed by four large projection neurons with cell bodies in the abdominal ganglia A1-2. The processes of these BAR-IR neurons are distributed dorsally and laterally in each of the ventral nerve cord ganglia. When baratin (10(-6)-10(-4) M) was applied to desheathed abdominal ganglia of locusts and cockroaches, we could monitor bursts of action potentials in neurons with axons in the anterior abdominal nerve (nerve 1), but not in the posterior nerve (nerve 2). In ganglia displaying spontaneous rhythmic firing in units of nerve 1, baratin strengthened the rhythmic pattern. Thus baratin appears to have a role in modulation of motor patterns in abdominal ganglia. The immunocytochemical findings suggest further modulatory actions of baratin in different circuits of the brain and ventral nerve cord. PMID- 10842232 TI - Peripheral nerve injury leads to the establishment of a novel pattern of sympathetic fibre innervation in the rat skin. AB - Peripheral nerve injury has been shown to result in sympathetic fibre sprouting around dorsal root ganglia (DRG) neurons. It has been suggested that this anomalous sympathetic fibre innervation of the DRG plays a role in neuropathic pain. Other studies have suggested an interaction between sympathetic and sensory fibres more peripherally. To date, no anatomical study of these possible interactions in the terminal fields of sensory and sympathetic fibres has been performed; therefore, the authors set out to study them in the rat lower lip after bilateral lesions of a sensory nerve, the mental nerve (MN). Immunocytochemistry for both substance P (SP) and dopamine-beta-hydroxylase (DbetaH) was performed. Within the first week post-MN lesions, the SP immunoreactive (IR) fibres had degenerated almost completely, whereas DbetaH-IR fibres were found in the upper dermis, an area from which they normally are absent. These DbetaH-IR fibres were present in the upper dermis at all postsurgery times studied (1, 2, 3, 4, 6, and 8 weeks). It is noteworthy that, although, by week 6 post-MN lesions, SP-IR fibre reinnervation of the lower lip was occurring, the DbetaH-IR fibres still were present in the upper dermis. Quantification revealed that the migration and branching of the DbetaH-IR fibres into the upper dermis occurred gradually and was most significant at 4 weeks post MN lesions, as demonstrated by the fact that the DbetaH-IR fibres were found 169.6 +/- 91.4 microm away from the surface of the skin compared with 407.1 +/- 78.4 microm away in sham-operated animals. These findings suggest that the ectopic innervation of the upper dermis by sympathetic fibres may be important in the genesis of neuropathic pain through the interactions of sympathetic and SP containing sensory fibres. PMID- 10842233 TI - The emergence of compartmental organization in olfactory bulb glomeruli during postnatal development. AB - The olfactory bulb glomerulus is a discrete and heterogeneous neuropil where olfactory receptor cell axons synapse with dendrites of mitral, tufted, and periglomerular neurons. To understand better the maturation of glomeruli and the spatiotemporal interactions that occur during postnatal development, we employed confocal microscopy and markers for immature and mature olfactory receptor cell axons in parallel with a marker for synaptic structure in maturing glomeruli. Sprague-Dawley rats at postnatal days 1, 6, 12, and 18 were processed for single- and double-label immunocytochemistry for olfactory marker protein (OMP), growth associated protein (GAP-43), and synaptophysin. Mature or adult-like subcompartmental organization within the glomerulus emerged by postnatal day 12. Earlier in development immature axons entered the core of the glomerulus and moved to the periphery as they matured. However, beginning around 12 days postnatal, immature axons distributed in the periphery and moved toward the core as they matured. This change in the trajectories of axons into glomeruli suggests that different rules may be followed in establishing versus maintaining glomeruli. Double labeling with OMP and synaptophysin demonstrated strong colocalization compared with GAP-43 and synaptophysin, which showed much less colocalization, consistent with the notion that OMP is associated with more mature axons. PMID- 10842234 TI - Propriospinal afferent and efferent connections of the lateral and medial areas of the dorsal horn (laminae I-IV) in the rat lumbar spinal cord. AB - The different subdivisions along the mediolateral extent of the superficial dorsal horn of the spinal cord are generally regarded as identical structures that execute the function of sensory information processing without any significant communication with other regions of the spinal gray matter. In contrast to this standing, here we endeavor to show that neural assemblies along the mediolateral extent of laminae I-IV cannot be regarded as identical structures. After injecting Phaseolus vulgaris leucoagglutinin and biotinylated dextran amine into various areas of the superficial dorsal horn (laminae I-IV) at the level of the lumbar spinal cord in rats, we have demonstrated that the medial and lateral areas of the superficial dorsal horn show the following distinct features in their propriospinal afferent and efferent connections: 1) A 300- to 400-microm-long section of the medial aspects of laminae I-IV projects to and receives afferent fibers from a three segment long compartment of the spinal dorsal gray matter, whereas the same length of the lateral aspects of laminae I IV projects to and receives afferent fibers from the entire rostrocaudal extent of the lumbar spinal cord. 2) The medial aspects of laminae I-IV project extensively to the lateral areas of the superficial dorsal horn. In contrast to this, the lateral areas of laminae I-IV, with the exception of a few fibers at the segmental level, do not project back to the medial territories. 3) There is a substantial direct commissural connection between the lateral aspects of laminae I-IV on the two sides of the lumbar spinal cord. The medial part of laminae I-IV, however, does not establish any direct connection with the gray matter on the opposite side. 4) The lateral aspects of laminae I-IV appear to be the primary source of fibers projecting to the ipsi- and contralateral ventral horns and supraspinal brain centers. Projecting fibers arise from the medial subdivision of laminae I-IV in a substantially lower number. The findings indicate that the medial and lateral areas of the superficial spinal dorsal horn of rats may play different roles in sensory information processing. PMID- 10842235 TI - Disruption of synaptic transmission or clock-gene-product oscillations in circadian pacemaker cells of Drosophila cause abnormal behavioral rhythms. AB - To study the function of clock-gene-expressing neurons, the tetanus-toxin light chain (TeTxLC), which blocks chemical synaptic transmission, was expressed under the control of promoters of the clock genes period (per) and timeless (tim), each fused to GAL4-encoding sequences. Although TeTxLC did not affect cycling of a clock-gene product at the gross level, it disrupted the rhythmic behavior of adult Drosophila. In constant darkness, the proportion of rhythmic flies was reduced in flies expressing active TeTxLC compared to controls, including those expressing inactive toxin. The behavior of TeTxLC-expressing flies was less synchronized to light:dark cycles than that of controls. To determine which neurons are responsible for these effects on behavior, the toxin was also expressed in restricted subsets of per/tim-expressing, laterally located pacemaker neurons by expressing TeTxLC under the control of a driver in which GAL4-encoding sequences are fused to the promoter of the pigment dispersing factor (pdf) gene. pdf-gal4-driven TeTxLC expression had relatively little effect on behavioral rhythms, implying that per/tim neurons other than pdf-expressing lateral neurons participate in the generation of rhythmic behavior. In another set of experiments, period gene products were expressed under the control of per gal4 or tim-gal4. This resulted in an increased level of PER protein in many brain cells and reduction of bioluminescence cycling reported by a per-luciferase transgene, especially in the case of per expression affected by tim-gal4. This indicates a disruption of the transcriptional feedback loop that is a part of the oscillatory mechanism underlying Drosophila's circadian rhythms. Consistent with this molecular defect, the proportion of rhythmic individuals in constant darkness was subnormal in flies expressing PER under the control of tim-gal4, and their behavior in light:dark cycles was abnormal. PMID- 10842236 TI - Apoptosis during sexual differentiation of the bed nucleus of the stria terminalis in the rat brain. AB - The bed nucleus of the stria terminalis (BST) in the rat forebrain differs between males and females. To test whether apoptosis may contribute to the development of sex differences in the BST, the incidence of apoptosis was determined in sham-treated males and sham-treated females sacrificed on postnatal days (PN) 2, 4, 6, 8, 10, and 12 (PN 1 being day of birth). More apoptotic nuclei were found in the principal nucleus of the BST (BSTpr) in females than in males, whereas the reverse was true for the lateral division of the BST (BSTl). Moreover, the volume of the BSTpr was larger in males than in females, whereas there was no sex difference in the volume of the BSTl. Our results also confirmed earlier reports indicating that the incidence of apoptosis in the central part of the medial preoptic nucleus (MPNc) is higher in females than in males. No sex difference in apoptosis was found in the ventromedial hypothalamus (VMH) and paraventricular nucleus (PVN). The volume of the MPNc and VMH was larger in males than in females, whereas the PVN volume did not differ between males and females. To test whether sex differences in neonatal levels of gonadal steroids may cause sex differences in the incidence of apoptosis in the BSTpr, the incidence of apoptosis was compared between castrated males and females that were treated with testosterone propionate or vehicle on the day of birth. In the BSTpr of gonadal steroid-treated animals, the incidence of apoptosis was lower when compared to animals treated with vehicle, which was also true for the MPNc. These results are consistent with the hypothesis that gonadal steroids contribute to the sexually dimorphic differentiation of the BST by controlling the incidence of apoptosis. PMID- 10842237 TI - Effects of captivity and testosterone on the volumes of four brain regions in the dark-eyed junco (Junco hyemalis). AB - This study investigates the effects of captivity and testosterone treatment on the volumes of brain regions involved in processing visual and spatial information in adult dark-eyed juncos (Junco hyemalis). We treated captive and free-living male juncos with either testosterone-filled or empty implants. Captive juncos had a smaller hippocampal formation (HF) (both in absolute volume and relative to telencephalon) than free-living birds, regardless of hormone treatment. Testosterone-treated males (both captive and free-living) had a smaller telencephalon and nucleus rotundus, but not a smaller HF or ectostriatum, than controls. We found that free-living testosterone-treated males had larger home ranges than free-living controls in agreement with earlier experiments, but we found no corresponding difference in HF volume. We discuss the implications of the effect of captivity on HF volume for past and future laboratory experiments. PMID- 10842238 TI - Seasonal expression of androgen receptors, estrogen receptors, and aromatase in the canary brain in relation to circulating androgens and estrogens. AB - Songbirds have a complex neural network for learning and production of song, namely the neural song system. Several nuclei of the song system contain androgen receptors (AR), and the neostriatal nucleus HVc also contains alpha type estrogen receptors (ER). Many songbird species show seasonal changes in both song and the neural song system that are correlated with seasonal variations in the circulating levels of gonadal steroids. However, there is increasing evidence that the sensitivity of the song system to gonadal steroids also changes seasonally. This could involve changes in the expression and activity of steroid receptors and steroid-metabolizing enzymes, such as the estrogen-synthesizing enzyme aromatase (AROM). The seasonal regulation of brain AR, ER, and AROM has not been studied before in the same individual songbirds. In this work, we compared plasma levels of androgens and estrogens, the expression level of AR-, ER-, and AROM-mRNA in the telencephalon, and brain AROM activity in male canaries between autumn (November) and spring (April) periods of high singing activity. Plasma levels of androgens and estrogens were higher in April than in November. The expression level of ER in HVc was higher in November than in April. In contrast, the expression level of AROM in the caudomedial neostriatum was higher in April than in November. However, we found no seasonal differences in the level of expression of AR and the volume of HVc as delimited by AR expression. Thus, AR expression in HVc was not correlated with circulating androgen levels. This study shows that both steroid-dependent and -independent seasonal factors regulate the action of gonadal hormones on the song system. In addition, we report a new site of AROM expression in the songbird brain, the nucleus interfacialis. PMID- 10842239 TI - Estradiol and progesterone regulate the expression of insulin-like growth factor I receptor and insulin-like growth factor binding protein-2 in the hypothalamus of adult female rats. AB - Gonadal hormones interact with insulin-like growthfactor-I (IGF-I) to regulate synaptic plasticity during the estrous cycle in the rat mediobasal hypothalamus. It has been proposed that tanycytes, specialized glial cells lining the ventral region of the third ventricle, may regulate the availability of IGF-I to hypothalamic neurons. IGF-I levels in tanycytes fluctuate during the estrous cycle. Furthermore, estrogen administration to ovariectomized rats increases IGF I levels in tanycytes, while progesterone, injected simultaneously with estrogen, blocks the estrogen-induced increase of IGF-I levels in tanycytes. To test whether hormonal regulation of IGF-I receptor (IGF-IR) and IGF binding protein-2 (IGFBP-2) may be involved in the accumulation of IGF-I in tanycytes, we assessed the effect of ovarian hormones on the levels of these molecules in the mediobasal hypothalamus of adult female rats. Ovariectomized animals were treated with either oil, estrogen, progesterone, or estrogen and progesterone simultaneously and then killed 6 or 24 h later. Some neurons, some astrocytes, and many tanycytes in the mediobasal hypothalamus were found by confocal microscopy to be immunoreactive for IGF-IR. IGFBP-2 immunoreactivity was restricted almost exclusively to tanycytes and ependymal cells and was colocalized with IGF-IR immunoreactivity in tanycytes. By electron microscope immunocytochemistry using colloidal gold labeling, IGF-IR and IGFBP-2 immunoreactivities were observed in the microvilli of tanycytes in the lumen of the third ventricle. IGF-IR and IGFBP 2 immunoreactive levels on the apical surface of tanycytes were significantly decreased by the administration of progesterone, either alone or in the presence of estradiol. IGF-IR levels in the mediobasal hypothalamus, measured by Western blotting, were not significantly affected by the separate administration of estradiol or progesterone to ovariectomized rats. However, the simultaneous administration of both hormones resulted in a marked decrease in IGF-IR protein levels. Estradiol administration to ovariectomized rats increased IGFBP-2 immunoreactive levels in the hypothalamus. While progesterone did not significantly affect IGFBP-2 expression, the simultaneous injection of estradiol and progesterone resulted in a marked decrease in IGFBP-2 protein levels. The effect of estradiol on IGFBP-2 was observed both in protein and mRNA levels, suggesting a transcriptional regulation. However, the simultaneous administration of progesterone and estradiol had different effects on IGF-IR protein and IGF-IR mRNA levels, as well as on IGFBP-2 protein and IGFBP-2 mRNA levels, suggesting a postranscriptional action. These findings indicate that estradiol and progesterone regulate the expression of IGF-IR and IGFBP-2 in the mediobasal hypothalamus of adult female rats. Regulation of the hypothalamic IGF-I system by ovarian hormones may be physiologically relevant for neuroendocrine regulation and for synaptic plasticity during the estrous cycle. These results do not support the hypothesis that estrogen-induced accumulation of IGF-I by tanycytes is mediated by the hormonal regulation of IGF-IR. However, estrogen-induced up regulation of IGFBP-2 and progesterone-induced down-regulation of IGF-IR and IGFBP-2 levels in the apical plasma membrane of tanycytes may be involved in the fluctuation of IGF-I levels in the mediobasal hypothalamus during the estrous cycle. PMID- 10842240 TI - The inositol 1,4,5-trisphosphate receptor is required for maintenance of olfactory adaptation in Drosophila antennae. AB - A role for inositol 1,4,5-trisphosphate (IP(3)) as a second messenger during olfactory transduction has been postulated in both vertebrates and invertebrates. However, given the absence of either suitable pharmacological reagents or mutant alleles specific for the IP(3) signaling pathway, an unequivocal demonstration of IP(3) function in olfaction has not been possible. Here we have investigated the role of a well-established cellular target of IP(3)-the IP(3) receptor (IP(3)R) in olfactory transduction in Drosophila. For this purpose we tested existing viable combinations of IP(3)R mutant alleles, as well as a newly generated set of viable itpr alleles, for olfactory function. In all of the viable allelic combinations primary olfactory responses were found to be normal. However, a subset of itpr alleles (including a null allele) exhibit faster recovery after a strong pulse of odor, indicating that the IP(3)R is required for maintenance of olfactory adaptation. Interestingly, this defect in adaptation is dominant for two of the alleles tested, suggesting that the mechanism of adaptation is sensitive to levels of the IP(3)R. PMID- 10842241 TI - Differential effects of the trophic factors BDNF, NT-4, GDNF, and IGF-I on the isthmo-optic nucleus in chick embryos. AB - The isthmo-optic nucleus (ION) of chick embryos is a model system for the study of retrograde trophic signaling in developing CNS neurons. The role of brain derived neurotrophic factor (BDNF) is well established in this system. Recent work has implicated neurotrophin-4 (NT-4), glial cell line-derived neurotrophic factor (GDNF), and insulin-like growth factor I (IGF-I) as additional trophic factors for ION neurons. Here it was examined in vitro and in vivo whether these factors are target-derived trophic factors for the ION in 13- to 16-day-old chick embryos. Unlike BDNF, neither GDNF, NT-4, nor IGF-I increased the survival of ION neurons in dissociated cultures identified by retrograde labeling with the fluorescent tracer DiI. BDNF and IGF-I promoted neurite outgrowth from ION explants, whereas GDNF and NT-4 had no effect. Injections of NT-4, but not GDNF, in the retina decreased the survival of ION neurons and accelerated cell death in the ION. NT-4-like immunoreactivity was present in the retina and the ION. Exogenous, radiolabeled NT-4, but not GDNF or IGF-I, was retrogradely transported from the retina to the ION. NT-4 transport was significantly reduced by coinjection of excess cold nerve growth factor (NGF), indicating that the majority of NT-4 bound to p75 neurotrophin receptors during axonal transport. Binding of NT-4 to chick p75 receptors was confirmed in L-cells, which express chick p75 receptors. These data indicate that GDNF has no direct trophic effects on ION neurons. IGF-I may be an afferent trophic factor for the ION, and NT-4 may act as an antagonist to BDNF, either by competing with BDNF for p75 and/or trkB binding or by signaling cell death via p75. PMID- 10842242 TI - Chemical and thermal stimuli have short-lived effects on the retzius cell in the medicinal leech. AB - During the appetitive phase of feeding, hungry leeches detect a prey by the integration of signals perceived by different sensory systems. Earlier reports suggested that chemical or thermal sensory stimulation of the lip was associated with increased afferent activity in cephalic nerves connecting the lip to the central nervous system. These authors further suggested that this activity was relayed to Retzius cells in segmental ganglia, which then released serotonin to initiate and control all aspects of feeding behavior. In this study, we show that chemosensory or thermal activation of the lip lasting for at least 5 min produces a distinct signal in the cephalic nerves consisting of action potentials of low amplitude. These small amplitude signals are clearly distinguishable from the large action potentials evoked by mechanosensory stimuli applied to the same area of the lip. Both types of sensory stimuli also evoke an increase in the firing frequency of the Retzius cells in segmental ganglia. However, the response recorded in the nerves and the Retzius cells during a maintained stimulus is not constant but decreases with an exponential time course. These results agree with our earlier observations on a semi-intact feeding preparation in which we showed that the firing frequency of the Retzius cell decreased as soon as the leech began to ingest its meal. Therefore, our data provide further evidence suggesting that it is unlikely that heat or chemical cues maintain the Retzius cell in an active state throughout the consummatory phase of feeding. PMID- 10842243 TI - A shift in focus. PMID- 10842244 TI - Expression of vascular endothelial growth factor (VEGF) and VEGF receptors in human neuroblastomas. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) is a specific endothelial cell mitogen that stimulates angiogenesis and plays a crucial role in tumor growth. The aim of the present study was to evaluate the expression of VEGF and of its two high-affinity tyrosine kinase receptors (KDR and Flt-1) in neuroblastoma surgical samples and cell lines. PROCEDURE: The VEGF, KDR, and Flt 1 mRNA expression in neuroblastoma surgical samples and cell lines was studied by RT-PCR. The receptors were identified in [(125)I]VEGF binding and in functional studies (effect on cell growth). VEGF production by neuroblastomas was investigated by the ELISA method. RESULTS: It was possible to observe the mRNAs encoding for VEGF and its two receptors in some of the surgical specimens examined, including most of the high-grade tumors. It was also possible to demonstrate that the SK-N-BE cell line expressed VEGF, KDR, and Flt-1 mRNAs as well as biologically active receptors: The cells bound [(125)I]-VEGF, and their growth was stimulated by exogenous VEGF. Moreover, VEGF protein could be detected in their culture conditioned medium. CONCLUSIONS: These results suggest that, in addition to its effect on angiogenesis, VEGF may affect neuroblastoma cell growth directly and could be an autocrine growth factor. PMID- 10842245 TI - Neuronal differentiation in human neuroblastoma cells by nerve growth factor following TrkA up-regulation by interferon-gamma. AB - BACKGROUND: TrkA mRNA expression has been reported to be related to favorable outcome of neuroblastoma (NB). Previously, we found that interferon-gamma (IFN gamma) can enhance TrkA mRNA expression in NB cell lines. In the present study, we examined the effect of nerve growth factor (NGF) on IFN-gamma-induced TrkA protein to clarify the relationship between TrkA and cell differentiation of NB. PROCEDURE: The effect of IFN-gamma on the TrkA mRNA expression was screened in six human NB cell lines and a freshly prepared sample, SK-rib, from a stage IV patient. Using two of them, we examined their morphological change during simultaneous loading of NGF and IFN-gamma. Tyrosine phosphorylation pattern after 5 min of NGF stimulation was also examined in immunoblot analysis with anti gp140(trkA) antibody and antiphospho tyrosine antibody. RESULTS: After a 4-day treatment with 500 IU/ml IFN-gamma, TrkA mRNA increased in five cell lines and SK rib cells in association with growth inhibition. Although the degree of morphological differentiation did not increase in proportion to the TrkA expression induced by IFN-gamma, continuous loading of both IFN-gamma and NGF caused marked morphological differentiation in a cultured KP-N-RT cell line and SK-rib cells during 10 days. Moreover, 5 min of NGF stimulation after IFN-gamma treatment caused the phosphorylation of TrkA protein and downstream proteins. CONCLUSIONS: IFN-gamma could induce the functional NGF receptor even in the aggressive phenotype of NB. PMID- 10842246 TI - Delayed craniospinal irradiation for a first isolated central nervous relapse of acute lymphoblastic leukemia: report on 14 cases. AB - BACKGROUND: Children developing an isolated central nervous system (CNS) relapse as first recurrence of their acute lymphoblastic leukemia (ALL) are considered to have a systemic relapse as well. They are mostly treated with intensive chemotherapy and craniospinal irradiation. In most treatment schedules, irradiation is given early after induction treatment. Because craniospinal irradiation affects a large portion of hematopoietic bone marrow systemically, treatment is often delayed owing to aplasias. Also, dose reductions are frequently needed. Children receiving simultaneously irradiation and chemotherapy are prone to (often severe) neurotoxicity. This study reports on children with a first isolated CNS relapse of their ALL receiving chemotherapy for 40 weeks. Treatment ends with the administration of irradiation given after cessation of chemotherapy. PROCEDURE: Fourteen children, with blasts and >5 cells/mm(3) in two consecutive samples of cerebrospinal fluid and a blast percentage <5% in their bone marrow were treated according to an intensive systemic and site-specific chemotherapy. Craniospinal irradiation was administered after cessation chemotherapy. RESULTS: Event-free-survival was 57% (confidence interval 35-89%), freedom from relapse was 61.5%; follow-up ranges from 2.0 to 15.1 years (median 11.7 years). One child died from septicemia during induction. Five children experienced a second relapse and died from their malignancy. Two children [with a t(9;22) or a rearranged MLL gene] relapsed prior to radiotherapy. Outcome was related to duration of first remission, age at relapse, and identification as a high-risk patient at initial diagnosis. No neurologic complications were noted during and after treatment. CONCLUSIONS: Delayed irradiation for isolated CNS relapse in children with ALL gives favorable survival rates, without significant toxicity. Neurotoxicity was absent. PMID- 10842247 TI - Biphenotypic sarcoma with characteristics of both a Ewing sarcoma and a desmoplastic small round cell tumor. AB - BACKGROUND: The EWS gene, a transcription factor of unknown function, is involved in chromosomal translocations associated with a wide variety of tumors, particularly small round blue cell tumors such as Ewing sarcoma. It has previously been reported that desmoplastic small round blue cell tumor (DSRBCT) frequently has an associated t(11;22) abnormality resulting from fusion of the EWS and WT-1 genes. PROCEDURE: We report a case of a small round blue cell tumor with characteristics of both Ewing sarcoma and DSRBCT with a t(11;22) translocation leading to fusion of the EWS and FLI1genes. RESULTS: The translocation point and fusion products were confirmed by polymerase chain reaction amplification and restriction fragment mapping of the products. CONCLUSIONS: The biphenotypic nature of this case and the apparent promiscuity of the EWS gene in tumor-associated translocations coupled with other reports of biphenotypic childhood sarcomas has potential implications for the relationship between small round blue cell tumors and the mechanism of EWS/FLI1 oncogenesis. PMID- 10842248 TI - Late effects of therapy in 94 patients with localized rhabdomyosarcoma of the orbit: Report from the Intergroup Rhabdomyosarcoma Study (IRS)-III, 1984-1991. AB - BACKGROUND: We reviewed the late complications of therapy in 94 patients with localized, primary rhabdomyosarcoma of the orbit treated on the Intergroup Rhabdomyosarcoma Study (IRS)-III protocol (1984-1991). PROCEDURE: A questionnaire was sent to the institutions that had registered 106 patients with orbital RMS on the IRS-III protocol, seeking information about vision, periocular structures, and growth and development of the 102 survivors. RESULTS: Ninety-four questionnaires were returned. The median follow-up interval was 7.6 years. The affected eye was removed from 13 patients because of local recurrence (N = 10) or other causes (N = 3). Seventy-nine of the eighty-one remaining patients had received radiation therapy. Sixty-five of these seventy-nine patients (82%) developed a cataract, and 43 of them (66%) underwent cataract surgery. Fifty-five patients (70%) had decreased visual acuity. Twenty-four patients had a dry eye, and 22 had chronic keratitis, conjunctivitis, or corneal changes. Strabismus, diplopia, retinopathy, and uveitis were uncommon. The orbit was hypoplastic in 48 of 82 patients assessed (59%). Ptosis and enophthalmos were reported in 22 patients. Decreased statural growth was noted in 13 of the 53 irradiated patients aged 3-14 years at diagnosis with sufficient data (24%). CONCLUSIONS: The overall survival rate was 96% (102/106). The eye was preserved in 86% of the patients, but vision was impaired in 70% of them. Other frequent complications were cataract, orbital hypoplasia, keratoconjunctivitis, and ptosis/enophthalmos. The current IRS-V study recommends decreasing the dose of irradiation and using conformal techniques in an attempt to minimize these complications. PMID- 10842249 TI - Cytotoxic chemotherapy has minimal direct effect on gastric myoelectric activity in children with 5HT(3) antagonist prophylaxis. AB - BACKGROUND: Cancer patients receiving cytotoxic chemotherapy frequently develop nausea and vomiting. The direct effect of chemotherapy on gastric pacemaker is not clear. The objective of this study was to assess the direct interference of gastric electrical activity by chemotherapeutic agents as a possible cause of vomiting using electrogastrography (EGG). PROCEDURE: Fasting surface EGGs were recorded in 24 children with malignancy and 24 age-matched controls. All oncology patients received the intravenous prophylatic ondansetron. According to the known emetic potential of the medications they received, the children were divided into mild, moderate, and severe groups. The EGG recordings of oncology patients were segmented into prechemotherapy period, chemotherapy period, and emetic period. The EGG data from each period were collected and analyzed (paired t-test and Mann Whitney U test). RESULTS: There were 24 children (M:F ratio: 14:10), with a mean age of 9.6 years. Eight children vomited during the course of recording (0/3 from the mild group, 2/6 from the moderate group, and 6/15 from the severe group). The average duration of intravenous medication infusion was 7.8 hr (range 2 min to 24 hr). There was no difference between the EGG of normal control and the prechemotherapy EGG of the oncology patients. No difference was detected between the EGG data from the prechemotherapy period and the chemotherapy period. A statistically significant increase in tachygastria was detected in emetic periods (28 episodes, mean duration 5 min; P = 0.01). CONCLUSIONS: Insignificant EGG changes during chemotherapy suggest that chemotherapy has a minimal direct effect on gastric pacemaker. Tachygastria seemed to be a secondary effect of vomiting rather than the cause of it. These preliminary findings suggest that efforts to modify gastric electrical rhythm as a means of management of chemotherapy-induced vomiting may be futile. Further studies to identify factors responsible for vomiting are warranted. PMID- 10842250 TI - The surgeon and the child with cancer: A report of the International Society of Pediatric Surgical Oncology (IPSO). PMID- 10842252 TI - Commentary PMID- 10842251 TI - Wilms tumor: does tumorectomy leave neoplastic tissue residual? PMID- 10842253 TI - Vitamin K prophylaxis and childhood cancer. PMID- 10842254 TI - Biological stability of RNA isolated from RNAlater-treated brain tumor and neuroblastoma xenografts. AB - BACKGROUND: Advances in molecular biological research have led to identification of prognostic factors such as Trk mRNA expression in primitive neuroectodermal tumors of the CNS and neuroblastoma. To study prospectively the importance of these prognostic factors in large groups of homogeneously treated patients, tumor specimens of good quality must be acquired, preserved, and stored at multiple institutions. Immediate freezing of tumor biopsy samples in liquid nitrogen and storage at -70 degrees C are the most commonly used method of tissue preservation for future RNA analysis. PROCEDURE: To evaluate alternative methods of preserving tissue samples for subsequent RNA analysis, we tested a new RNA stabilization solution. Using tumor tissue of two CNS tumor and one neuroblastoma human xenografts, we compared total RNA isolated from tumor tissue stored for 7 days at room temperature in stabilization solution to that of snap-frozen tissue. The quality of the RNA was studied by spectrophotometry, gel electrophoresis, RT-PCR, and gene expression profiling. RESULTS: No major differences were observed in the quality of RNA isolated from tumor samples stored at room temperature in the RNA stabilization solution compared to snap-frozen tumor samples stored at -70 degrees C. CONCLUSIONS: High-quality RNA can be prepared from tumor tissue stored at room temperature. Whenever snap freezing is not feasible, pieces of tumor tissue can be treated with RNAlater for subsequent RNA analysis. Short-term storage and shipment of well-preserved tumor tissue are clearly feasible for all institutions, thereby facilitating large multiinstitutional studies of biological prognostic factors. PMID- 10842255 TI - Adult respiratory distress syndrome (ARDS) after chemotherapy. PMID- 10842256 TI - Ewing sarcoma of the clavicle in a 10-month-old patient. PMID- 10842257 TI - Congenital sacrococcygeal primitive neuroectodermal tumor. PMID- 10842258 TI - Prognostication matters. PMID- 10842259 TI - Triplet repeat expansion in neuromuscular disease. AB - Expansions of unstable trinucleotide repeats cause at least 15 inherited neurologic diseases. Here we review what has been learned of three neuromuscular diseases caused by this type of mutation. X-linked spinal and bulbar muscular atrophy is a motor neuronopathy caused by a CAG repeat expansion in the androgen receptor gene. The mutated protein has an expanded polyglutamine tract, forms intranuclear aggregates, and mediates neurodegeneration through a toxic gain-of function mechanism. Oculopharyngeal muscular dystrophy is a dominantly inherited myopathy caused by a GCG/polyalanine expansion in the gene encoding poly(A) binding protein 2. Myotonic dystrophy is a clinically variable multisystem disease caused by a CTG expansion in the 3' untranslated region of the myotonin gene. For each of these disorders, we summarize the clinical and pathologic features and review current understanding of the molecular mechanisms underlying their pathogenesis. PMID- 10842260 TI - Skeletal muscle response to tenotomy. AB - Tenotomy is a commonly encountered clinical entity, whether traumatic or iatrogenic. This article reviews the response of skeletal muscle to tenotomy. The changes are subdivided into molecular, architectural, and functional categories. Architectural disruption of the muscle includes myofiber disorganization, central core necrosis, Z-line streaming, fibrosis of fibers and Golgi tendon organs, changes in sarcomere number, and alterations in the number of membrane particles. Molecular changes include transient changes in myosin heavy chain composition and expression of neural cell adhesion molecule (NCAM). Functionally, tenotomized muscle produces decreased maximum tetanic and twitch tension. Alterations in normal skeletal muscle structure and function are clinically applicable to the understanding of pathological states that follow tendon rupture and iatrogenic tenotomy. PMID- 10842261 TI - Traumatic injury to peripheral nerves. AB - This article reviews the epidemiology and classification of traumatic peripheral nerve injuries, the effects of these injuries on nerve and muscle, and how electrodiagnosis is used to help classify the injury. Mechanisms of recovery are also reviewed. Motor and sensory nerve conduction studies, needle electromyography, and other electrophysiological methods are particularly useful for localizing peripheral nerve injuries, detecting and quantifying the degree of axon loss, and contributing toward treatment decisions as well as prognostication. PMID- 10842263 TI - Peripheral neuropathy in chronic venous insufficiency. AB - Chronic venous insufficiency (CVI) of the lower legs may cause tissue damage, but involvement of peripheral nerves is uncertain. We examined 30 patients with CVI and 20 healthy controls using motor and sensory nerve conduction studies, vibration testing and thermotesting, quantitative sudomotor axon-reflex test, and laser Doppler flowmetry. Subjects with possible confounding factors for peripheral neuropathies were excluded. Prolongation of distal motor latency of the peroneal nerve (median, 5.4 versus 4.5 ms; P = 0.02), increased limits for warm (9.60 degrees C versus 5.20 degrees C; P = 0.016) and cold detection (3.45 degrees C versus 1.55 degrees C; P = 0.016) and reduced vibration sense (2.8925 versus 1.1075; P < 0.008) were found. The results demonstrate a disturbance of A alpha fibers, A-beta fibers, A-delta fibers, and thermoafferent-C fibers, possibly induced by ischemia due to venous microangiopathy and increased endoneurial pressure. Analogous to neuropathic ulcers in diabetes, the CVI associated neuropathy may also be a cofactor in the development of venous ulcers. PMID- 10842262 TI - Linear estimates of disease progression predict survival in patients with amyotrophic lateral sclerosis. AB - We have shown that linear estimates of rates of disease progression (LEP), derived from isometric myometry [grip or foot dorsiflexion (FD) strength] and forced vital capacity (FVC%), are clinically and statistically significant predictors of survival of patients with amyotrophic lateral sclerosis (ALS) from date of disease onset and, except those based on grip strength, of survival from the date of measurement. We tested these results in 2 additional groups of patients: 1) those who participated in a previously reported Protropin (GH) study; and 2) those enrolled in two other clinical trials (group 2). The LEP were derived and tested as predictors of survival. In a Cox proportional hazards model, LEP based on all measures predicted survival from disease onset in both groups of patients. Using cutoff points determined within the original group to stratify patients in the validation groups into faster and slower progressing subgroups resulted in statistically significant separation of survival curves from disease onset in group 2 for all LEP and in group 1 (the GH group) for LEP derived from FD strength; and, for survival from date of measurement in group 2, when stratified by LEP based on FD strength or FVC%. LEP based on data generated by myometry or pulmonary function studies have now been shown to predict survival in 3 unrelated groups of patients with ALS entering clinical trials. Their precise use in clinical trial design needs to be explored further. PMID- 10842264 TI - Some observations on fibrillations and positive sharp waves. AB - Electromyographic recordings of fibrillation potentials (FPs) and positive sharp waves (PSWs) demonstrate transformation of FP to PSW and vice versa, atypical firing patterns, changes in waveform shape and amplitude, and time-locked potentials. The etiology of the waveform characteristics of FP and PSW is discussed based on abnormal propagation in a small section of muscle fiber that is "damaged" by the needle. The results of simple computer simulations are described. PMID- 10842265 TI - Anti-SGPG antibody in CIDP: nosological position of IgM anti-MAG/SGPG antibody associated neuropathy. AB - Polyneuropathy with monoclonal gammopathy usually is considered a nosological entity different from chronic inflammatory demyelinating polyneuropathy (CIDP). Criteria proposed by the American Academy of Neurology AIDS Task Force (1991), however, show monoclonal gammopathy to be a condition concurrent with CIDP. The purpose of this study was to clarify the nosological relationship between CIDP and IgM anti-myelin-associated glycoprotein (MAG)/sulfated glucuronyl paragloboside (SGPG)-associated polyneuropathy. We investigated IgM anti-MAG/SGPG antibody in 85 CIDP patients by various methods, then examined the relation of M protein to the presence of IgM anti-MAG/SGPG antibody. In our large study, 17 (20%) of 85 CIDP patients had high IgM anti-SGPG antibody titers in the enzyme linked immunosorbent assay. This was confirmed by thin-layer chromatography immunostaining for IgM anti-SGPG antibody and immunoblotting for IgM anti-MAG antibody. Immunoelectrophoresis and immunofixation, respectively, detected IgM M protein in 6 (35%) and 13 (76%) of the 17 CIDP patients. We conclude that some patients with IgM anti-MAG/SGPG antibody with or without monoclonal gammopathy may be diagnosed as having CIDP, when patients are diagnosed according to the current CIDP criteria. PMID- 10842266 TI - Natural history of 46 patients with multifocal motor neuropathy with conduction block. AB - We studied 46 consecutive patients with multifocal motor neuropathy with conduction block (MMN-CB). Typically, asymmetric weakness and atrophy of the hands or arms developed insidiously, but spontaneous improvement (without treatment) or death from this disease did not occur and 94% remained employed. For 18 patients examined on multiple occasions using the weakness subscore of the neuropathy impairment score [NIS(W)] for a median time of 2.3 years, worsening of 1.3 points per year was observed; many patients, however, had received intensive immunomodulating therapy. Median worsening to our first evaluation (generally without treatment) was estimated at 4.2 points per year, perhaps suggesting that treatment had influenced course. Three criteria for conduction block (CB) were compared, but the least stringent was sensitive for the diagnosis. Conduction block accompanied by weakness and atrophy typically affected only motor fibers, especially of midforearm nerves, and these sites of dysfunction persisted for months or years. Neurological signs and electrodiagnostic features were consistent with CB, axonal degeneration, a variable degree of reinnervation, and segmental demyelination. Although this study did not focus on therapy, intravenous gammaglobulin and cyclophosphamide appeared to be associated with neurological improvement, which was seldom complete or sustained. Axonal degeneration and faulty regeneration may in part explain this muted response. Possibly, treatment must be earlier, more intense, or different. PMID- 10842267 TI - Central fatigue during isometric exercise in amyotrophic lateral sclerosis. AB - While both upper and lower motor neuron dysfunction may contribute to impaired muscle function in amyotrophic lateral sclerosis (ALS), the precise mechanisms of muscle fatigue have not been clarified in this disease. Therefore, the central and peripheral factors in muscle fatigue were investigated during intermittent submaximal isometric ankle dorsiflexion in 7 patients with ALS and 6 healthy control subjects. Voluntary and electrically stimulated force, central and peripheral indices of muscle activation, and intramuscular energy metabolism were measured before and during exercise. At the end of exercise, only the ALS group had an increase in the "added force" in response to a stimulus train imposed during maximal voluntary contraction, indicating significant central fatigue in ALS. In support of this conclusion, patients with ALS had less intramuscular phosphocreatine depletion and less fatigue of stimulated tetanic force during exercise compared to control. Thus, due to the central failure, there was decreased muscle activation resulting in a smaller metabolic demand and less fatigue within the muscle itself. These data demonstrate a major contribution of central factors to muscle fatigue in ALS. PMID- 10842268 TI - Phrenic nerve conduction in infancy and early childhood. AB - Diaphragmatic action potentials (DAPs) were mapped on the thorax bilaterally in 16 neurologically normal infants and 8 boys aged 1 to 4 years during artificial ventilation after thoracic surgery. Transcutaneous stimulation was used to activate the phrenic nerve at the supraclavicular fossa at the end of an artificial inspiration. The DAPs were of positive polarity and were recorded on the ipsilateral anterolateral chest wall over the sixth to the eighth intercostal spaces, with a maximal peak at the seventh intercostal space. The DAP latencies gradually decreased from 6 to 8 ms at birth to about 5 ms at the age of 1 year, despite an increase of conduction distance. Statistical analyses revealed that DAP amplitude did not correlate with age. The latencies and amplitudes of the DAPs displayed little interside variation. The results are valuable not only as a reference for the diagnosis of patients with phrenic nerve palsy, but also as an indicator of the normal development of the phrenic nerve. PMID- 10842269 TI - Functional magnetic stimulation facilitates gastrointestinal transit of liquids in rats. AB - The purpose of this study was to investigate the effect of a relatively novel technology, functional magnetic stimulation (FMS), on gastrointestinal transit of liquids in rats. Orogastric gavage with technetium-99 solution was used to assess gastric emptying and gastrointestinal transit time in 92 rats. FMS was performed over the anterior cervical and/or dorsal thoracolumbar regions using a figure-8 coil. Stimulation protocols were 1, 2, or 4 h in length. FMS accelerated gastric emptying and decreased gastrointestinal transit time. The acceleration was dependent on the stimulation parameters used as well as on the duration of the protocol; high levels of FMS produced a quicker effect, whereas lower levels were effective at later times. This study provides evidence that FMS could be an alternative or adjunct therapy to treat disorders in gastrointestinal motility. PMID- 10842270 TI - Posture-related changes of soleus H-reflex excitability. AB - We investigated whether the modulatory effects of segmental and descending inputs on the soleus H reflex are modified by postural conditions. Fourteen healthy volunteers received a transcranial magnetic stimulus (TMS) or percutaneous electrical stimulation of the posterior tibial nerve (PTN), preceding by 0 to 400 ms the elicitation of the soleus H reflex in supine, sitting, and standing positions. In all positions, TMS induced an early period of facilitation at interstimulus intervals (ISIs) ranging between 5 and 35 ms. In supine and sitting positions, there was a second period of facilitation at ISIs between 60 and 90 ms, which was absent or significantly reduced in the standing position. PTN induced a strong inhibition of the H reflex in all positions up to 125 ms. In supine and sitting positions, inhibition continued up to 400 ms, whereas it was significantly reduced or completely absent beyond 125 ms in the standing position. These results demonstrate posture-related differences in the modulatory effects of descending and segmental inputs on the excitability of the H-reflex circuit. PMID- 10842271 TI - A prospective study of handcuff neuropathies. AB - Prior reports of handcuff-related neuropathies have been retrospective or small series of cases selected for their unusual electrophysiological or historical features. We conducted a prospective 27-month study at a large, urban, teaching hospital of all patients with a complaint of hand numbness, weakness, or paresthesias attributed to overtightened handcuffs. Forty-one patients were evaluated clinically, and electrodiagnostic testing was performed on 18. In the group with electrodiagnostic testing, neuropathies due to overtightened handcuffs were detected in 22 superficial radial, 12 median, and 9 ulnar nerves. The correlation between clinical and electrodiagnostic findings was best for superficial radial neuropathies, some of which were severe. Median and ulnar nerve injuries were generally mild. Similar clinical findings were obtained in the group without electrodiagnostic testing. Handcuff-related injury to the most commonly affected nerve, the superficial radial, can be severe and permanent. PMID- 10842272 TI - Trigemino-facial reflex inhibitory responses in some lower facial muscles. AB - The effects of electrical trigeminal stimulation on activated facial muscles were studied in 20 normal subjects in order to evaluate whether excitatory or inhibitory responses are present and to investigate whether the reflex organization is similar in all the facial muscles. No inhibition was observed in frontalis, orbicularis oculi, orbicularis oris, and mentalis muscles. By contrast, a clear suppression of electromyographic (EMG) activity (late silent period or SP2) was present in the levator labii superioris, depressor anguli oris, and depressor labii inferioris muscles, with a mean latency ranging from 41.8 to 50.2 ms, and a mean duration ranging from 27.5 to 40.9 ms. An early suppression of EMG activity (early silent period or SP1) was observed, with a latency of 16 to 20 ms and a duration of 10 ms, mainly in inferior perioral muscles. Our findings show a selective trigeminal inhibitory influence upon some specific lower facial muscles. PMID- 10842274 TI - Quantitative features of the stretch response of extrinsic finger muscles in hemiparetic stroke. AB - Despite its potential importance in hand dysfunction, spasticity in the finger muscles following stroke has not been well described. To explore this area, we assessed the role of finger flexor spasticity, along with that of passive mechanical forces, in resisting finger movement in 13 chronic stroke subjects. Subjects were tested with a device that stretched the extrinsic finger muscles through imposed rotation of the metacarpophalangeal (MCP) joints. Both maintained and constant-velocity stretches were imposed. For the constant-velocity stretches, eight of the 13 stroke subjects exhibited strong stretch reflexes, as determined by electromyography and net work. The net work of this reflex response, calculated from the integral of the torque-angle plots, increased proportionally with increasing velocity, indicating a contribution from flexor muscle spasticity. Conversely, nine of the 13 stroke subjects did not possess distinctly greater passive, mechanical resistance to MCP rotation than control subjects. While extensor spasticity was not observed, stretch of the extrinsic finger flexors also produced some reflex activity in the finger extensors concomitant with reflex excitation of the flexors. These findings suggest that resistance to muscle stretching following stoke is mediated primarily by neurological rather than biomechanical disturbances, although changes in muscle fiber length may exaggerate the resistance. PMID- 10842273 TI - Altered expression of costimulatory molecules in myasthenia gravis. AB - To characterize the involvement of costimulatory pathways in the pathogenesis of myasthenia gravis (MG), a multiparameter flow cytometry assay was adopted to enumerate blood mononuclear cells (MNC) expressing CD28, CD80, CD86, CD40, and CD40L molecules in patients with MG and healthy subjects. Patients with MG had lower percentages of CD8(+)CD28(+) cells, augmented percentages of CD4(+)CD80(+), CD4(+)CD86(+), CD8(+)CD80(+), CD8(+)CD86(+), CD14(+)CD80(+), and CD14(+)CD86(+) cells, and similar levels of cells expressing CD40 and CD40L and of B cells expressing CD80 and CD86 compared to the controls. Patients with early onset of MG (<40 years) had lower percentages of CD3(+)CD86(+), CD4(+)CD86(+), CD8(+)CD86(+) T cells and CD20(+)CD86(+) B cells compared to those with late onset (>40 years). There was a positive correlation between the patients' age and percentages of CD86(+) cells. The data indicate that the CD28/CD80-CD86 costimulatory pathway is involved in MG. The high percentages of CD80 and CD86 positive T cells and monocytes may reflect persistent activation of T and B cells, whereas the low CD28 expression may be the result of chronic exposure to CD80 and CD86. These molecules could be the focus for new and improved immunomodulating therapies of MG. PMID- 10842275 TI - Increased levels of leukemia inhibitory factor mRNA in muscular dystrophy and human muscle trauma. AB - Leukemia inhibitory factor (LIF) is an important muscle trauma factor both after crush injury and in the mdx mouse dystrophy model. It is important to establish which growth factors have a role in human muscle regeneration due to potential clinical therapeutic applications. As there is limited information concerning LIF expression in human muscle, we investigated the relative levels of LIF messenger ribonucleic acid (mRNA) in human muscle injury. Semiquantitative reverse transcriptase followed by polymerase chain reaction was used to amplify LIF message. We found that although LIF mRNA is expressed in low levels in control muscle, a sevenfold increase occurred after orthopedic muscle trauma and a marked 19-fold increase in dystrophic muscle (P < 0.002). These results indicate that LIF mRNA is upregulated in surgical and especially medical muscle injury with repeated myonecrosis. Muscle growth factors such as LIF may assist in future muscle rehabilitation after injury. PMID- 10842276 TI - Induction of the ATP-sensitive potassium (uK(ATP)-1) channel by endotoxemia. AB - The effect of sepsis on the ubiquitously expressed ATP-sensitive potassium (uK(ATP)-1) channel expression was measured in Sprague-Dawley rat diaphragms. Rats were treated with either 0.5 ml saline or 20 mg/Kg E. coli lipopolysaccharides and sacrificed at 3, 6, 12, 24, or 48 h later. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis showed that channel mRNA expression was increased at 3 h and continued to rise up to 48 h. Western blotting analysis showed a approximately 9-fold increase in channel protein expression 24 h after sepsis. Our results demonstrate that sepsis upregulates the uK(ATP)-1 channel. PMID- 10842277 TI - Prevalence of sporadic inclusion body myositis in Western Australia. AB - A 10-year retrospective review was conducted to ascertain the prevalence of inclusion body myositis (IBM) in Western Australia. Seventeen patients with sporadic IBM aged 45-90 years were identified and the prevalence of IBM was calculated to be 9.3 x 10(-6). The prevalence was higher in men (10.9 x 10(-6)) than in women (7.7 x 10(-6)). The mean age of onset of IBM was 56.6 years, and the mean delay between onset of symptoms and diagnosis was 4.4 years. The age adjusted prevalence over the age of 50 years was 35.3 x 10(-6). The results suggest a higher prevalence of IBM than has previously been reported. PMID- 10842278 TI - The effect of leg extension training on the mean power frequency of the mechanomyographic signal. AB - The purpose of the present investigation was to examine the effect of concentric isokinetic leg extension training on the mean power frequency (MPF) of the mechanomyographic (MMG) signal. Twenty-one men were assigned into a training (TRN; n = 12) or control (CTL; n = 9) group. The TRN group performed six sets of leg extensions 3 days per week for 12 weeks at a velocity of 90 degrees /s. All subjects were tested every 4 weeks for peak torque (PT), while MMG was recorded from the vastus lateralis. PT increased, but there was no significant (P > 0.05) change in the MMG MPF over the 12-week training period. These results indicate that MMG MPF, measured from the vastus lateralis, was not sensitive to training induced increases in leg-extension strength, possibly due to competing influences of hypertrophy on the MMG signal and/or training-induced adaptations in muscles other than the vastus lateralis. PMID- 10842279 TI - The motor tinel sign: a useful sign in entrapment neuropathy? AB - The motor Tinel sign (MTS) refers to electromyographic activity, sometimes associated with a visible muscle jerk, evoked by percussion or manipulation of a peripheral nerve. The MTS is found in entrapment neuropathies (such as carpal tunnel syndrome or entrapment of the ulnar nerve at the elbow and peroneal nerve at the fibular head) but occasionally also in normal subjects. The MTS may be useful in evaluating patients with entrapment neuropathy, but it does not always indicate nerve dysfunction. PMID- 10842280 TI - Fulminant case of hereditary neuropathy with liability to pressure palsy. AB - Hereditary neuropathy with liability to pressure palsy (HNPP) is typified as isolated nerve palsies caused by trivial compression or trauma. It rarely presents in two extremities and even more infrequently affects all four limbs simultaneously. We present a patient who concurrently experienced right shoulder, left hand, and bilateral foot weakness mimicking several multifocal conditions. Electromyography suggested HNPP and subsequent nerve biopsy and genetic testing were confirmatory. The case demonstrates that HNPP can present in a fulminant manner and should be included in the differential diagnosis of acute multiple mononeuropathies. The possible causes for such a rapid clinical course in our patient are discussed. PMID- 10842281 TI - Partial alpha-sarcoglycan deficiency with retention of the dystrophin glycoprotein complex in a LGMD2D family. AB - In patients with sarcoglycan (SG) deficiency, a primary defect in any one of the four SG proteins usually leads to reduced expression of the whole SG complex. We report a limb-girdle muscular dystrophy type 2D family (LGMD2D), with variable phenotype, where a mutation in the alpha-SG gene resulted in the partial deficiency of alpha-SG alone. The normal expression of the other three SG proteins suggests that mutations close to the alpha-SG transmembrane domain might be less critical for complex integrity, and that weakness may occur despite its retention. PMID- 10842282 TI - Plantar muscle fibrillations and positive sharp waves. PMID- 10842284 TI - AAEM news and comments PMID- 10842283 TI - Blink reflex in Behcet's disease. PMID- 10842286 TI - Physical characteristics of young boys with fragile X syndrome: reasons for difficulties in making a diagnosis in young males. AB - Fragile X syndrome is the leading form of hereditary mental retardation, but the condition is still underdiagnosed in young children. Because of concern that the fragile X phenotype is subtle in young boys and therefore contributes to underdiagnosis of the disorder, we evaluated 73 boys (36 with fragile X and 37 same-age boys who were fragile X negative) using a checklist that we devised to learn which characteristics might be the most useful for alerting professionals to this diagnosis. After a multiple comparisons adjustment, only 4 of 42 characteristics differed significantly in their distributions between the two groups of boys (P < 0.0012), but 10 other items may also have predictive value for fragile X syndrome (P < 0.01). Four additional items occurred in at least 80% of boys with fragile X and may also be helpful for the clinician. Professionals who work with developmentally delayed children should be aware of these 18 clinical characteristics and some of the behavior characteristics commonly seen in boys with fragile X so that they can readily diagnose patients. PMID- 10842287 TI - Three novel TBX5 mutations in Chinese patients with Holt-Oram syndrome. AB - Holt-Oram syndrome (HOS) is an autosomal dominant syndrome that comprises upper limb and cardiac defects. The gene responsible for HOS, TBX5, was isolated and many mutations have been identified in HOS patients. We analyzed 11 Chinese HOS patients (7 from three families and 4 sporadic cases) for TBX5 mutation by single strand conformation polymorphisms (SSCPs). Three SSCP changes were detected in two of the three familial cases and one sporadic case. Sequence analysis identified three novel, heterozygous mutations in TBX5: a frameshift mutation caused by one base deletion [C416del] in one family, a mis-sense mutation (Gln49Lys) induced by a base substitution (C145A) in another family, and the other mis-sense mutation (Ile54Thr) by T161C in one sporadic case. The patients with the frameshift mutations had severer clinical manifestations that involved aplasia/hypoplasia of the arm and thumbs, while those with the mis-sense mutations presented with milder anomalies such as absent or hypoplastic thumbs but without arm abnormalities. These observations may support a genotype phenotype correlation in HOS patients with TBX5 mutation. PMID- 10842288 TI - Mutations in the MEFV gene in a large series of patients with a clinical diagnosis of familial Mediterranean fever. AB - Familial Mediterranean fever (FMF) is an autosomal recessively inherited disease affecting patients of the Mediterranean basin. FMF is characterized by recurrent episodes of fever accompanied with topical signs of inflammation. Some patients can develop a renal amyloidosis associated (AA) amyloidosis. The administration of colchicine is an effective preventive treatment of both the attacks and amyloidosis. The FMF gene (MEFV) was cloned and missense mutations were found to be responsible for the disease. We investigated a large series of 303 unselected and unrelated patients of various ethnic backgrounds with a clinical suspicion of FMF to confirm or invalidate the diagnosis of FMF and to determine the spectrum of MEFV mutations. Molecular analysis focused on all the most frequent mutations identified so far, and an exhaustive analysis of exon 10, containing the mutational hotspot, was performed through DNA sequencing. Sixty-two percent of Sephardic, North African Arabs, Armenian and Turkish patients were either homozygous or compound heterozygous for MEFV mutations. In other populations surrounding the Mediterranean Sea such as Greek, Italian, Portuguese, Kurdish and Lebanese populations, mutations were also found. In general, patients without Mediterranean origin had no mutations in the MEFV gene. Two new mis-sense mutations were identified in exon 10 of the MEFV gene: the S675N in an Italian patient and the M680L in a French patient without any known at-risk ethnic ancestry. PMID- 10842289 TI - Familial Mediterranean fever in two Bedouin families: mutation analysis and disease severity. AB - Familial Mediterranean fever (FMF) is an autosomal recessive disease prevalent among non-Ashkenazi Jews, Armenians, Arabs, and Turks. The Bedouin are nomad Arab tribes residing in desert margins of the Middle East and Arabia. FMF is quite rare in Bedouins, and here we report on two Bedouin families from southern Israel suffering from this disorder. The MEFV mutations found in the Bedouin patients M694I, V726A, and E148Q are consistent with their Arab origin. The disease severity score showed a mild to moderate severity disease in six patients. The Bedouins, leading a unique nomadic life, may prove instrumental in unraveling the role of environmental factors in the course and severity of FMF. PMID- 10842290 TI - Detection of a cryptic translocation in a family with mental retardation using FISH and telomere region-specific probes. AB - Cryptic rearrangements involving the telomeres are thought to account for a substantial number of patients with unexplained mental retardation and multiple congenital anomalies, although the exact incidence of these rearrangements is still unclear. With the advent of chromosome-specific telomeric probes and the use of FISH (fluorescence in situ hybridization), it is now possible to identify submicroscopic rearrangements of the distal ends of chromosomes that may otherwise go undetected using conventional cytogenetic studies. We report on a 4 1/2 year-old girl with severe mental retardation and minor anomalies who inherited the unbalanced product of a cryptic translocation involving chromosomes 2 and 17 from her father. The family history was significant for early pregnancy losses, stillbirths, and mental retardation in many other family members, suggesting segregation of a familial translocation. This translocation was detected using chromosome-specific telomere FISH probes, and not visible using conventional cytogenetic methods. Collectively, this case and those previously reported clearly demonstrate the value of a systematic search for cryptic chromosome rearrangements in patients with unexplained mental retardation with previously reported normal chromosome studies; and in particular those with a family history of mental retardation, birth defects, or early pregnancy losses. PMID- 10842291 TI - Structural analysis of a rare rearranged Y chromosome and its bearing on genotype phenotype correlation. AB - We report on a 9-year-old boy with a rare rearranged Y chromosome and borderline short stature (-2.0 SD). Standard metaphase chromosome analysis indicated a 46,X,i(Y)(q1O) karyotype, but high resolution G-banding showed an asymmetric band pattern for the rearranged Y chromosome. FISH and DNA studies for a total of 15 different Y chromosomal loci or regions showed that the rearranged Y chromosome was accompanied by: 1) a partial deletion of the short arm pseudoautosomal region (PAR1) involving SHOX, with the breakpoint distal to DXYS85; and 2) a partial duplication of Yq, with the breakpoint proximal to DAZ. The karyotype was determined as 46,X,?i(Y)(q1O).ish der(Y)(Yqter--> Yp11.3::Yq11.2- >Yqter)(DAZ++,DYZ3+,SRY +, SHOX-). The X chromosome and the autosomes were normal. The results suggest that haploinsufficiency of SHOX is primarily responsible for the borderline short stature, and that the deletion of the PAR1 may result in spermatogenic failure due to defective X-Y pairing and recombination in the PAR1. PMID- 10842292 TI - Maternal uniparental disomy (UPD) for chromosome 2 discovered by exclusion of paternity. AB - Serological and molecular (DNA-STR) analysis of a paternity case demonstrated exclusion of paternity of the presumptive father in two markers (ACP and Apo B, both localized on chromosome 2, region 2p25.2 and 2p23/24, respectively) in a phenotypically normal girl with a normal karyotype 46,XX (by GT-banding). The index of paternity calculated for other serological (seven erythrocyte antigens, six serum protein systems, and seven isozymes, as well as the A- and B-HLA loci) and nine DNA markers, excluding ACP and Apo B, gives a very high (virtually certain) degree of paternity for the presumptive father. Maternal uniparental disomy (UPD) for chromosome 2 was suspected. Evaluation of polymorphic DNA markers (STRs) spanning chromosome 2 of the child, mother, and presumptive father demonstrated that the girl had inherited two maternal chromosome 2 homologues, whereas alleles for markers from other chromosomes were inherited from the father in a Mendelian fashion. The girl was homoallelic for informative markers mapping to the chromosomal regions 2p23-25, but she was heteroallelic for informative markers on the long arm of chromosome 2, establishing that the maternal UPD with partial isodisomy of the short arm was caused by a meiosis I nondisjunction event with genetic recombination (chiasmata in this region 2p23-25) during oogenesis. PMID- 10842293 TI - Breakpoint within the nucleolus organizer region resulting in a reciprocal translocation t (4;14)(q21;p12). AB - Reciprocal translocations involving a break in the nucleolus organizer region (NOR) are rare. A balanced translocation in a mother and her fetus with breakpoints in the NOR at 14p12 and on the long arm of a chromosome 4 at band 4q21 is described. The rearrangement was characterized by Ag-NOR staining, multiplex fluorescence in situ hybridization (M-FISH), and FISH with rDNA probes. This and other cases with breakpoints within NORs are discussed. PMID- 10842295 TI - New case of Cole-Carpenter syndrome. AB - We describe a girl with a severe progressive type of osteogenesis imperfecta, in association with multisutural craniosynostosis, growth failure, and craniofacial findings including ocular proptosis, marked frontal bossing, midface hypoplasia, and micrognathia. Collagen analysis was normal. These features are consistent with the diagnosis of Cole-Carpenter syndrome. This report provides further evidence for the existence of this rare genetic entity. PMID- 10842294 TI - Frequent association of 22q11.2 deletion with tetralogy of Fallot. AB - Chromosome 22q11.2 deletion causes DiGeorge syndrome, velocardiofacial syndrome, conotruncal anomaly face syndrome with tetralogy of Fallot (TOF), and sporadic or familial TOF. To determine the prevalence and clinical importance of the 22q11.2 deletion in TOF, a series of 212 Japanese TOF patients was studied. The type of pulmonary blood supply, which may lead to various clinical outcomes, and other additional anomalies were evaluated clinically. The 22q11.2 deletion was diagnosed by fluorescence in situ hybridization with N25 and TUPLE1 probes. Of the 212 patients examined, 28 (13%) had a 22q11.2 deletion, the frequency being higher than that in TOF patients with trisomy 21. The prevalence of the deletion in TOF patients with pulmonary atresia (PA) plus major aortico-pulmonary collateral arteries (MAPCA) was significantly higher than the value in patients with PA plus patent ductus arteriosus (PDA) (P = 0.04) or with pulmonary stenosis (PS) (P < 0.0001). All 28 patients with 22q11.2 deletion had one or more extracardiac abnormalities. Four of 9 patients with the 22q11.2 deletion and TOF PA-MAPCA suffered from bronchomalacia, while none of 19 patients with TOF-PA-PDA or TOF-PS manifested bronchomalacia (P = 0.006). These results indicate that 22q11.2 deletion is the most frequent cause of syndromic TOF, especially for TOF PA-MAPCA, and bronchomalacia is the clinically most important associated anomaly in TOF-PA-MAPCA patients. PMID- 10842296 TI - Colobomatous microphthalmia, microcephaly with cerebellar hypoplasia: association or new syndrome? AB - We report on a 3.5-year-old girl with microcephaly, microphthalmia, coloboma of the iris, mild developmental delay, and other minor anomalies. Neuroimaging showed marked cerebellar and vermian hypoplasia. This condition has not been described previously and is discussed in the context of the "micro syndrome," together with other similar syndromes. Our case highlights the heterogeneity of the "microphthalmia plus brain malformations" group of patients. PMID- 10842297 TI - Comparative genomic hybridization: a new approach to screening for intrauterine complete or mosaic aneuploidy. AB - In the practice of clinical genetics chromosomal aneuploidy in both mosaic and nonmosaic forms has long been recognized as a cause of abnormal prenatal and postnatal development. Traditionally, cytogenetic analysis of cultured lymphocytes has been used as a standard test for detection of constitutional aneuploidies. As lymphocytes represent only one lineage, chromosomal mosaicism expressed in other tissues often remains undetected. The purpose of this study was to assess the utilization of molecular cytogenetic analysis for detection of chromosomal aneuploidy in placental tissues. Using placentas from 100 pregnancies with viable nonmalformed livebirths, both trophoblast and chorionic stroma were analyzed using comparative genomic hybridization (CGH). In all cases with an indication of chromosomal imbalance by CGH, fluorescence in situ hybridization (FISH) analysis was performed to confirm the presence of aneuploidy. To differentiate between constitutional aneuploidy and confined placental mosaicism (CPM), amniotic membrane was analyzed by CGH and FISH techniques. Our results demonstrated five placentas with CPM for chromosomes 2, 4, 12, 13, and 18, respectively, and two constitutional nonmosaic aneuploidies (47,XXX and 47,XXY). Molecular cytogenetic studies of human placental tissues enables easy analysis of both embryonic (amnion) and extraembryonic (chorion) cell lineages. Detection at birth of chromosomal defects affecting intrauterine placental and fetal development is important because these chromosomal defects may continue to have an influence on postnatal development. PMID- 10842299 TI - Population and molecular genetic update: the second Middle East Genetics Association of America (MEGA) conference. PMID- 10842298 TI - Homozygosity mapping in a family with microcephaly, mental retardation, and short stature to a Cohen syndrome region on 8q21.3-8q22.1: redefining a clinical entity. AB - A syndrome of microcephaly, progressive postnatal growth deficiency, and mental retardation was observed in two brothers and their cousin from a multiply consanguineous kindred of Lebanese descent. Hypotonia, chorioretinal dystrophy, and myopia were also identified. The severity of the condition varied among the closely related patients. Because of absence of a distinctive facial appearance, the degree of mental retardation, and short stature, the initially considered clinical diagnosis of Cohen syndrome was withdrawn and a novel genetic entity was assumed. Homozygosity mapping in this family assigned the gene to a 26.8-cM region on the chromosome band 8q21.3 -22.1, between the microsatellites at D8S270 and D8S514. The maximum two-point LOD score was found for marker at D8S267 (Zmax=3.237 at Omax=0.00). Intriguingly enough, the identified gene region overlaps the refined gene region for Cohen syndrome (COH1) [Kolehmainen et al., 1997: Euro J Hum Genet 5:206-213]. This fact encourages the hypothesis that the described kindred segregates for a variant of Cohen syndrome and suggests a redefinition of its phenotype. PMID- 10842300 TI - Regarding trisomy 2p syndrome. PMID- 10842301 TI - De novo partial duplication of long arm of chromosome 13: dup(13)(q12-->q14) PMID- 10842302 TI - Life, death and Sonic hedgehog. AB - The secreted glycoprotein Sonic hedgehog (SHH), a vertebrate homologue of the Drosophila segment polarity gene Hedgehog, is essential for the development of diverse tissues during embryogenesis. Studies of SHH function during neural tube and somite development have focused on its role in specifying the dorsoventral polarity of these structures, but a recent report by Ahlgren and Bronner Fraser(1) supports the possibility that SHH has additional functions in cell survival and cell proliferation. Perturbation of SHH signaling after the early dorsoventral specification of the cranial neural tube leads to increased cell death in both the neural tube and the neural crest. This implies that SHH is continually required as a trophic and/or mitogenic factor during brain development, and expands the variety of cellular responses to SHH signaling. BioEssays 22:499-502, 2000. PMID- 10842303 TI - Building a protein interaction map: research in the post-genome era. AB - With the extensive amount of information generated by genome-wide sequencing, the entire set of gene products in an organism can now be predicted. The challenge of understanding the function of each gene in the genome has led to the development of many large-scale and high-throughput experimental techniques. Recently, two papers, Walhout et al.(1) and Uetz et al.,(2) have described studies that add a new functional dimension to research conducted on a genome-wide scale. These two groups have utilized the yeast two-hybrid system to identify interactions among the entire complement of proteins encoded by the Caenorhabditis elegans and the Saccharomyces cerevisiae genomes, respectively. Using a set of 29 genes that have been previously characterized, Walhout et al. demonstrated the feasibility and efficiency of this technique by building an interaction matrix among a large number of proteins. On an even larger scale, Uetz et al. conducted two-hybrid analyses using proteins that represent over 87% of the total gene products in yeast and identified interactions for about 15% of the total yeast proteins. BioEssays 22:503-506, 2000. PMID- 10842304 TI - Toggles and oscillators: new genetic circuit designs. AB - Two recent papers report the de novo design of a functioning biological circuit using well-characterized genetic elements.(1,2) Gardner et al. designed and constructed a genetic toggle switch while Elowitz and Leibler built an oscillating genetic circuit. Both circuits were designed with the aid of mathematical models. These papers demonstrate progress towards the unification of theory and experiment in the study of genetic circuits. Comparison of the predicted and observed behavior of the circuits, however, shows that the models explain only some of the circuits' properties. Further study of the observed behaviors not predicted by the model would lead to new insight into the properties of genetic networks. BioEssays 22:507-509, 2000. PMID- 10842305 TI - Remodeling muscles with calcineurin. AB - Ca(2+) signaling plays a central role in hypertrophic growth of cardiac and skeletal muscle in response to mechanical load and a variety of signals. However, the mechanisms whereby alterations in Ca(2+) in the cytoplasm activate the hypertrophic response and result in longterm changes in muscle gene expression are unclear. The Ca(2+), calmodulin-dependent protein phosphatase calcineurin has been proposed to control cardiac and skeletal muscle hypertrophy by acting as a Ca(2+) sensor that couples prolonged changes in Ca(2+) levels to reprogramming of muscle gene expression. Calcineurin also controls the contractile and metabolic properties of skeletal muscle by activating the slow muscle fiber-specific gene program, which is dependent on Ca(2+) signaling. Transcription factors of the NFAT and MEF2 families serve as endpoints for the signaling pathways whereby calcineurin controls muscle hypertrophy and fiber-type. We consider these findings in the context of a model for Ca(2+)-regulated gene expression in muscle cells and discuss potential implications of these findings for pharmacologic modification of cardiac and skeletal muscle function. BioEssays 22:510-519, 2000. PMID- 10842306 TI - Post-transcriptional gene-silencing: RNAs on the attack or on the defense? AB - Post-transcriptional gene-silencing (PTGS) was first discovered in plants and results from the sequence-specific degradation of RNA. Degradation can be activated by introducing transgenes, RNA viruses or DNA sequences that are homologous to expressed genes. A similar RNA degradation mechanism which is inducible by double-stranded RNA (dsRNAs), has been discovered recently in vertebrates, invertebrates and protozoa. dsRNAs may also be potent activators of PTGS in plants. PTGS is not cell autonomous, suggesting the synthesis of sequence specific silencing signals which are not only moving through the plant but are also amplified and an RNA-directed RNA Polymerase which has recently been cloned from various plant species is a candidate enzyme for amplifying silencing signals. The natural role of PTGS seems to be as a defence against plant viruses, so what first appeared to be RNAs on the attack may now be considered RNAs on the defense. BioEssays 22:520-531, 2000. PMID- 10842307 TI - Nuclear targeting signal recognition: a key control point in nuclear transport? AB - Recent progress indicates that there are multiple pathways of nucleocytoplasmic transport which involve specific targeting sequences, such as nuclear localization sequences (NLSs), and cytosolic receptor molecules of the importin/karyopherin superfamily which recognise and dock the NLS-containing proteins at the nuclear pore. This first step of nuclear import/export is of central importance, with the affinity of the importin-targeting sequence interaction a critical parameter in determining transport efficiency. Different importins possess distinct NLS-binding specificities, which allows the system to be modulated through differential expression of the importins themselves, as well as through competition between different importins for the same protein, and between different proteins for the same importin. The targeting sequence-importin interaction can also be influenced directly by phosphorylation increasing the affinity of the interaction with importins or by targeting sequence masking through phosphorylation or specific protein binding. Targeting sequence recognition thus appears to represent a key control point in the regulation of nuclear transport. BioEssays 22:532-544, 2000. PMID- 10842308 TI - A look at the Caenorhabditis elegans Kex2/Subtilisin-like proprotein convertase family. AB - Significant advances have recently been made in our understanding of the mechanisms of activation of proteins that require processing. Often this involves endoproteolytic cleavage of precursor forms at basic residues, and is carried out by a group of serine endoproteinases, termed the proprotein convertases. In mammals, seven different convertases have been identified to date. These act in both the regulated secretory pathway for the processing of prohormones and proneuropeptides and in the constitutive secretory pathway, in which a variety of proproteins are activated endoproteolytically. The recently completed sequence of the nematode Caenorhabditis elegans genome affords a unique opportunity to examine the entire proprotein convertase family in a multicellular organism. Here we review the nature of the family, emphasising the structural features, characteristic of the four nematode genes, that supply all of the necessary functions unique to this group of serine endoproteinases. Studies of the C. elegans genes not only provide important information about the evaluation of this gene family but should help to illuminate the roles of these proteins in mammalian systems. BioEssays 22:545-553, 2000. PMID- 10842309 TI - Mammary gland neoplasia: insights from transgenic mouse models. AB - Current theories of breast cancer progression have been greatly influenced by the development and refinement of mouse transgenic and gene targeting technologies. Early transgenic mouse models confirmed the involvement of oncogenes, previously implicated in human breast cancer, by establishing a causal relationship between overexpression or activation of these genes and mammary tumorigenesis. More recently, the importance of genes located at sites of loss of heterozygosity in human breast cancer have been examined in mice by their targeted disruption via homologous recombination. The union of these two approaches allows the generation of complex animal models that more accurately reflect the multistep nature of human breast cancer. This review will examine how the study of transgenic mice has increased our understanding of the molecular events responsible for oncogenic transformation of the mammary gland. BioEssays 22:554-563, 2000. PMID- 10842310 TI - No sex please, we're mitochondria: a hypothesis on the somatic unit of inheritance of mammalian mtDNA. AB - In this article we develop a model for the organization and maintenance of mitochondrial DNA (mtDNA) in mammalian somatic cells, based on the idea that the unit of genetic function comprises a group of mtDNA molecules that are semi permanently associated as a mitochondrial nucleoid. Different mtDNA molecules within a nucleoid need not be genetically identical. We propose that nucleoids replicate faithfully via a kind of mitochondrial mitosis, generating daughter nucleoids that are identical copies of each other, but which can themselves segregate freely. This model can account for the very slow rates of mitotic segregation observed in cultured, heteroplasmic cell-lines, and also for the apparently poor complementation observed between different mutant mtDNAs co introduced into rho(0) cells (cells that lack endogenous mtDNA). It also provides a potential system for maintaining the mitochondrial genetic fitness of stem cells in the face of a presumed high somatic mutation rate of mtDNA and many rounds of cell division in the absence of phenotypic selection. BioEssays 22:564 572, 2000. PMID- 10842311 TI - Plant GRAS and metazoan STATs: one family? AB - GRAS is a recently discovered family of plant-specific proteins that play important regulatory roles in diverse aspects of plant development. Several of the motifs present in the GRAS proteins suggest that they function as transcription factors, although homology-searching programs have revealed no significant similarity to any non-plant proteins. Here we propose that the GRAS proteins are related to the Signal Transducers and Activators of Transcription (STAT) family of proteins. STATs are known in many non-plant species, and act as intracellular intermediaries between extracellular ligands and the transcription and activation of genes. Our hypothesis is that the GRAS proteins perform this function in plants, with mechanisms similar to those of the animal STATs. If true, this hypothesis has important implications for the evolution of phosphotyrosine based signal transduction systems in eukaryotic organisms. BioEssays 22:573-577, 2000. PMID- 10842312 TI - Regeneration in the metazoans: why does it happen? AB - Why does regeneration occur? And why, when it manifests itself, does it do so in some but not all metazoan species? Hence, what are the permissive or inhibitory factors operating behind this phenomenon? When it comes to regeneration, many questions, such as these, remain unanswered. In fact, the problem of animal regeneration has withstood the probing of scientific inquiry for over 250 years and still awaits a satisfactory mechanistic explanation. In this essay, I will review the distribution and the modes of regeneration that are found in the different metazoan phyla. Also, I will re-examine ideas on its evolutionary origins, and discuss its possible relationship to both asexual reproduction and embryogenesis. This endeavor has two objectives. First, to bring forward an interpretation of regeneration which integrates evolutionary and developmental considerations into its discussion. And second, to suggest a comparative experimental approach to this problem that may bring us closer to understanding the molecular basis of this long-standing biological problem. BioEssays 22:578 590, 2000. PMID- 10842313 TI - Recruitment of chromatin remodeling machines. AB - The assembly of eukaryotic DNA into folded nucleosomal arrays has drastic consequences for many nuclear processes that require access to the DNA sequence, including RNA transcription, DNA replication, recombination, and repair. Two types of highly conserved chromatin remodeling enzymes have been implicated as regulators of the repressive nature of chromatin structure: ATP-dependent remodeling complexes and nuclear histone acetyltransferases (HATs). Recent studies indicate that both types of enzymes can be recruited to chromosomal loci through either physical interactions with transcriptional activators or via the global accessibility of chromatin during S phase of the cell cycle. Here we review these recent observations and discuss the implications for gene-specific regulation by chromatin remodeling machines. PMID- 10842314 TI - Nonlymphoid cultured cells possess a system controlling cellular compatibility. AB - We show that various nonlymphoid cultured cells can activate the production of cytotoxic factors in response to direct contact with cells of a different kind. Accumulation of cytotoxic factors in the medium was detected 1 h after contact of K562 and L929 cells or after contact of L929 cells with purified membranes of K562 cells. TNF-alpha or immunologically related proteins, or both, but not Fas ligand or lymphotoxin, were also accumulated in membranes of K562 and L929 cells shortly after these cells had been allowed to contact each other. The cytotoxic factors expressed by nonlymphoid cells trigger apoptosis of target cells. These observations strongly suggest that nonlymphoid cells possess molecular mechanisms controlling cellular compatibility. PMID- 10842315 TI - Identification of multiple cis-acting elements mediating the induction of prostaglandin G/H synthase-2 by phorbol ester in murine osteoblastic cells. AB - The tumor promoter phorbol 13-myristate 12-acetate (PMA), the best characterized protein kinase C agonist, frequently regulates gene expression via activation of Fos/Jun (AP-1) complexes. PMA rapidly and transiently induces prostaglandin G/H synthase-2 (PGHS-2) expression in murine osteoblastic MC3T3-E1 cells, but no functional AP-1 binding motifs in the 5'-flanking region have been identified. In MC3T3-E1 cells transfected with -371/+70 bp of the PGHS-2 gene fused to a luciferase reporter gene (Pluc), PMA stimulates luciferase activity up to eightfold. Computer analysis of the sequence of the PGHS-2 promoter region identified three potential AP-1 elements in the -371/+70 bp region, and deletion analysis suggested that the sequence 5'-aGAGTCA-3' at -69/-63 bp was most likely to mediate stimulation by PMA. Mutation of the putative AP-1 sequence reduces the ability of PMA to stimulate Pluc activity by 65%. On electrophoretic mobility shift analysis (EMSA), PMA induces binding to a PGHS-2 probe spanning this sequence, binding is blocked by an unlabeled AP-1 canonical sequence, and antibodies specific for c-Jun and c-Fos inhibit binding. Mutation of this AP-1 site also causes a small (22%) but significant reduction in the serum stimulation of Pluc activity in transiently transfected MC3T3-E1 cells. On EMSA, serum induces binding to a PGHS-2 probe spanning the AP-1 site, binding is blocked by an unlabeled AP-1 canonical sequence, and antibodies specific for c-Jun and c-Fos inhibit binding. Joint mutation of this AP-1 site and the nearby CRE site at -56/ 52 bp, previously shown to mediate serum, v-src and PDGF induction of PGHS-2 in NIH-3T3 cells, blocks both PMA and serum induction of Pluc activity in MC3T3-E1 cells. Hence, the AP-1 and CRE binding sites are jointly but differentially involved in both the PMA and serum stimulation of PGHS-2 promoter activity. PMID- 10842316 TI - HOXB7 overexpression promotes differentiation of C3H10T1/2 cells to smooth muscle cells. AB - The presence of immature smooth muscle cells and ectopic tissues such as fully formed bone in atherosclerotic lesions, may result from recapitulation of embryonic mechanisms in the artery wall. We hypothesized that expression of homeobox genes is triggered in atherogenesis and that these regulate proliferation and differentiation of multipotential progenitor cells along one or more specific lineages. We identified expression of the homeobox gene HOXB7 in clones of bovine aortic medial cells previously shown to be multipotent. HOXB7 was subsequently detected in human atherosclerotic plaques by RT-PCR and in situ hybridization. Expression was localized to areas adjacent to calcification and scattered in media and neointima, which may be reflective of a role in either osteoblastic or smooth muscle cell differentiation. To differentiate between these possibilities, we overexpressed HOXB7 in C3H10T1/2 cells, a multipotent cell line able to differentiate into vascular smooth muscle cells (SMC), as well as osteogenic and chondrogenic lineages. Results showed that overexpression of HOXB7 increased proliferation 3.5-fold, and induced an SMC-like cell morphology. In addition, expression of the early SMC markers calponin and SM22alpha increased 4-fold and 3-fold respectively by semi-quantitative RT-PCR. Expression of the intermediate SMC marker smooth muscle myosin heavy chain (SM-MHC) did not change. No increase in osteogenic or chondrogenic differentiation was detected, neither in the C3H10T1/2 cells nor in M2 cells, a bone marrow stromal cell line used to confirm this result. These findings suggest that HOXB7 plays a role in expansion of immature cell populations or dedifferentiation of mature cells. PMID- 10842317 TI - Diacylglycerol mediates the T-cell receptor-driven Ca(2+) influx in T cells by a novel mechanism independent of protein kinase C activation. AB - The mechanism of Ca(2+) influx in nonexcitable cells is not known yet. According to the capacitative hypothesis, Ca(2+) influx is triggered by IP(3)-mediated Ca(2+) release from the intracellular Ca(2+) stores. Conversely, many workers have reported a lack of association between release and influx. In this work, the role of diacylglycerol (DAG) as the mediator of T-cell receptor (TCR)-driven Ca(2+) influx in T cells was investigated. Stimulation of mouse splenic T cells with naturally occurring DAG caused Ca(2+) entry in a dose- and time-dependent manner. Such stimulation was blocked by Ni(2+), a divalent cation known to block Ca(2+) channels. Inhibition of protein kinase C (PKC) by calphostin C did not inhibit, but slightly enhanced, the DAG-stimulated Ca(2+) entry. However, inhibition of DAG metabolism by DAG kinase and lipase inhibitors enhanced the DAG stimulated Ca(2+) entry. DAG lipase and kinase inhibitors also enhanced the Ca(2+) entry in T cells stimulated through TCR/CD3 complex with anti-CD3 antibody. Calphostin C did not affect the anti-CD3-stimulated Ca(2+) entry. These results showed that TCR-driven Ca(2+) influx in T cells is mediated by DAG through a novel mechanism(s) independent of PKC activation. PMID- 10842318 TI - Combined use of insulin and endothelin-1 causes decrease of protein expression of beta-subunit of insulin receptor, insulin receptor substrate-1, and insulin stimulated glucose uptake in rat adipocytes. AB - Previously, we reported that insulin-stimulated glucose uptake (ISGU) can be inhibited by endothelin (ET-1). However, the mechanism by which ET-1 impairs ISGU in adipocytes remains unclear. This study investigated the effects of ET-1 on insulin action in rat adipocytes in order to elucidate the molecular mechanism of action of ET-1 on ISGU. The results show that ISGU was increased fivefold after 3 h treatment with 1 nM insulin. Treatment with 100 nM ET-1 had no effect on basal glucose uptake. However, ET-1 inhibited approximately 25% of ISGU and 20% of insulin binding after 3-h treatment in the presence of 1 nM insulin. Expression of the beta-subunit of the insulin receptor (IRbeta) and the insulin receptor substrate-1 (IRS-1) in adipocytes was not significantly affected by 1 nM insulin or by 100 nM ET-1, even after 3-h treatment. However, expressions of IRbeta and IRS-1 were dramatically decreased in a dose- and time-dependent manner when adipocytes were treated with both insulin and ET-1. Approximately 50% of IRbeta and 65% of IRS-1 expression levels were suppressed when adipocytes were simultaneously treated with both 1 nM insulin and 100 nM ET-1 for 3 h. These results suggest that the inhibitory effect of ET-1 on ISGU may be mediated via the insulin receptor and suppression of IRbeta/IRS-1 expression. PMID- 10842319 TI - Janus kinase 2 is associated with a box 1-like motif and phosphorylates a critical tyrosine residue in the cytoplasmic region of cytotoxic T lymphocyte associated molecule-4. AB - It is a consensus that a cytotoxic T lymphocyte associated molecule-4 (CTLA-4) transduces inhibitory signal for T cell activation under physiological condition, indicating that this molecule is an important regulator of T cell homeostasis in vivo. It has been reported that phosphorylation and dephosphorylation of tyrosine residue Y-165 in the cytoplasmic region of CTLA-4 play an important role in its negative signaling and cell surface expression. Some signaling molecules such as Src homology 2 protein tyrosine phosphatase 2 (SHP-2) and the p85 subunit of phosphatidylinositol 3 kinase (PI3 kinase) associate with phosphorylated tyrosine residue Y-165, through Src homology 2 (SH2) domains. On the other hand, the adapter complex proteins, AP-2 and AP-50 interact with the same tyrosine residue when unphosphorylated, resulting in clathrin-mediated endocytosis of CTLA-4 molecules. The objective of this study is to identify a tyrosine kinase that can directly bind and phosphorylate the critical tyrosine residue, Y-165 in the cytoplasmic domain of CTLA-4. Here, we demonstrated that 1) Janus Kinase 2 (Jak2) was directly associated with a box 1-like motif in the cytoplasmic tail of CTLA-4 molecule, 2) Jak2 phosphorylated Y-165 residue in the cytoplasmic region of CTLA 4 molecule, and 3) Jak2 was associated with CTLA-4 in HUT 78 T cell lines. PMID- 10842320 TI - Binding motifs of CBP2 a potential cell surface target for carcinoma cells. AB - Previously we have shown (Hebert et al. [1999] J. Cell Biochem. 73:248-258) that among many cell lines the CBP2 gene product, Hsp47, eludes its retention receptor, erd2P, resulting in the appearance of Hsp47 on the cell surface associated with the tetraspanin protein CD9. Since Hsp47 possesses a highly restricted binding cleft, random peptide display libraries were used to characterize peptides binding to Hsp47 and then to target this protein on carcinoma cell lines in vitro. Comparison of the clones obtained from panning revealed little specific homology based on sequence alone. To determine whether carcinoma cells expressing Hsp47 could selectively take up the selected bacteriophages, traditional immunofluorescence and confocal microscopy were employed. These studies revealed that phage-displaying Hsp47 binding peptides bound to cell lines expressing Hsp47 and that the peptides were rapidly taken up to a location coincident with Hsp47 staining. These observations were confirmed by cytometric analyses. These data indicate that CBP2 product may provide a molecular target for chemotherapy and/or imaging of malignancies. PMID- 10842321 TI - Behavior of nucleolar proteins during the course of apoptosis in camptothecin treated HL60 cells. AB - By means of immunofluorescence and immunoelectron microscopy we have studied the fate of different nucleolar components during the apoptotic process in camptothecin-treated HL60 cells. We have found that RNA polymerase I disappeared while UBF was associated with previously described fibrogranular threaded bodies. In contrast, fibrillarin, C23/nucleolin, and B23/nucleophosmin remained detectable in granular material present amid micronuclei of late apoptotic cells. Double immunolabeling experiments showed colocalization of both C23 and B23 with fibrillarin. Immunoblotting analysis showed that UBF was proteolytically degraded, whereas fibrillarin, C23/nucleolin, and B23/nucleophosmin were not. These results may help explain the presence of anti-nucleolar antibodies seen in various pathological disorders. PMID- 10842322 TI - In vivo protein-DNA interactions at the kinin B(1) receptor gene promoter: no modification on interleukin-1 beta or lipopolysaccharide induction. AB - The kinin B(1) receptor (B(1)R) gene is strongly upregulated following tissue injury and inflammation. In an attempt to define the regulatory elements that account for the control of B(1)R gene expression, we have conducted in vivo footprinting analysis of the B(1)R gene promoter region in three human cell types: embryonic lung fibroblast cells (IMR-90), embryonic kidney cells (HEK 293), and primary cultures of vascular umbilical smooth muscle cells. Initial in vitro delineation of the B(1)R gene promoter by transient transfection experiments with a reporter gene indicated that a 1.4-kb region, located just upstream of the transcription initiation site, bears all the characteristics of a core promoter with a functional TATA box and additional positive and negative control elements, as some of them could be tissue-specific. In vivo ultraviolet and dimethylsulfate footprinting analyses of the 1.4-kb region revealed no difference between the footprint patterns in the three cell types studied. We found that even in the noninduced state, the B(1)R gene promoter is possibly bound by several sequence-specific DNA binding proteins (GATA-1, PEA3, AP-1, CAAT, Sp1, Pit-1a, Oct-1, CREB). Some other footprints were detected on sequences that do not correspond to any known transcription factor binding site. No additional changes in protein-DNA complexes were observed upon treatment with interleukin-1 beta (IL-1beta) or bacterial lipopolysaccharide, shown previously to induce B(1)R gene expression. These results indicate that complex protein-DNA interactions exist at the B(1)R gene promoter prior to induction by external stimuli even in cells (HEK-293) that do not express a functional B(1)R. PMID- 10842323 TI - Phosphatidic acid induces calcium influx in neutrophils via verapamil-sensitive calcium channels. AB - Phosphatidic acid (PA) induces a biphasic Ca(2+) mobilization response in human neutrophils. The initial increase is due to the mobilization of Ca(2+) from intracellular stores, whereas the secondary increase is due to the influx of Ca(2+) from extracellular sources. The present investigation characterizes PA induced Ca(2+) influx in neutrophils. Depolarization of neutrophils by 50 mM KCl enhanced PA-induced Ca(2+) influx, whereas verapamil, a Ca(2+) channel blocker, attenuated this response in a dose-dependent manner. These observations suggest that PA-induced Ca(2+) influx is mediated via verapamil-sensitive Ca(2+) channels. Stimulation of neutrophils with exogenous PA results in accumulation of endogenously generated PA with a time course similar to the effects of exogenous PA on Ca(2+) influx. Ethanol inhibited the accumulation of endogenous PA and calcium mobilization, indicating that activation of membrane phospholipase D plays a role in PA-mediated Ca(2+) influx. The results of this study suggest that exogenously added PA stimulates the generation of intracellular PA, which then mediates Ca(2+) influx through verapamil-sensitive Ca(2+) channels. PMID- 10842324 TI - Nuclear and cytoplasmic tau proteins from human nonneuronal cells share common structural and functional features with brain tau. AB - The heterogeneous family of tau proteins interacts with microtubules, actin filaments, and intermediate filaments. The tau isoforms have been shown to play a major role in neuronal polarity. However, tau-like proteins have been found in several other types of cells. Previous studies have also indicated the presence of a nuclear tau. The relationships between nuclear and cytoplasmic tau as well as the functional aspects of the nuclear tau are unknown. In this study, we demonstrate by reverse transcriptase polymerase chain reaction using specific primers that a transcript with features of neuronal tau is present in human fibroblast and Huh-7 hepatoma cell lines. Additionally, we present the first isolation and characterization of cytosolic and nuclear tau-like proteins from nonneuronal cells. Nonneuronal cytosolic tau components were isolated using the perchloric acid precipitation approach, while nuclear tau was isolated after selective extractions using high-ionic strength buffers. The cytoplasmic tau of nonneuronal cells is composed of at least three isoforms, whereas two main isoforms were detected in nuclear tau. Interestingly, the cytoplasmic and nuclear tau components exhibited the capacity to promote tubulin polymerization in vitro. Immunofluorescence studies using monoclonal anti-tau antibodies indicated a discrete distribution of tau protein in both the interphase and mitotic nucleus. In the latter, tau colocalized with the chromosomal scaffold. These studies, together with previous evidence on tau roles in modulating microtubule growth from centrosomes, and its role in the interaction patterns that stabilize the integrity of the cytoskeletal network, strongly support the idea that tau is a multifunctional protein involved in fundamental cellular processes. PMID- 10842325 TI - B lymphocytes inhibit human osteoclastogenesis by secretion of TGFbeta. AB - The role of B lymphocytes in osteoclast (OC) formation is controversial, because both stimulatory and inhibitory effects of B-lineage cells on osteoclastogenesis and life span have been reported. In this study, we have investigated the effects of mature B cells on human osteoclastogenesis using cultures of peripheral blood stem cells (PBSC), a system that generates functional OCs in the absence of stromal cells. We report that B cells inhibit the formation of OCs and shorten the life span of mature OCs by secreting transforming growth factor beta (TGFbeta), a factor that induces apoptosis in these cells. The antiosteoclastogenic effects of B cells are abolished by addition of anti-TGFbeta antibody to osteoclast cultures and mimicked by treatment of B cell-deprived PBSC cultures with recombinant TGFbeta, thus confirming TGFbeta as the B cell produced antiosteoclastogenic activity. Thus, the ability of B cells to downregulate osteoclastogenesis by secretion of the apoptotic cytokine TGFbeta provides new insights into the ability of immune cells to regulate OC formation under basal and inflammatory conditions. PMID- 10842326 TI - Effect of extracellular magnesium on topoisomerase II activity and expression in human leukemia HL-60 cells. AB - Topoisomerase II (TopoII) is a Mg-dependent enzyme involved in topological modifications of DNA that are crucial to the regulation of cell proliferation and possibly differentiation. To investigate the role of Mg availability in the modulation of TopoII in whole cells, we studied enzyme activity and expression in HL-60 cells grown in the presence of decreasing amounts of extracellular Mg (0.5, 0.03, and 0.01 mM MgSO(4)). In comparison to cells grown in 0.5 mM Mg, cells grown in 0.03 mM Mg exhibited a decrease in TopoII activity, as evidenced by reduced induction of DNA/TopoII cleavable complexes and apoptosis by etoposide and teniposide. Enzyme activity was restored by the readdition of Mg (0.5 and 1.5 mM) in the incubation medium, confirming that this effect was indeed modulated by extracellular Mg. Restriction of Mg to 0.01 mM was associated with a dramatic decrease in TopoII activity resembling that observed in HL-60 cells differentiated by dimethyl sulfoxide treatment. The restriction of Mg, while decreasing enzyme activity, was found to upregulate TopoII protein expression, determined by Western blot analysis. The increase of TopoII protein levels was correlative with the degree of Mg deprivation. Collectively, these results indicate that extracellular levels of Mg may control availability of intracellular Mg, thus affecting the regulation of TopoII activity/expression and downstream processes of cell proliferation and/or differentiation. PMID- 10842328 TI - Phosphorylation of tubulin tyrosine ligase: a potential mechanism for regulation of alpha-tubulin tyrosination. AB - The tubulin tyrosination/detyrosination cycle is a well-established posttranslational modification, which is carried out by two enzymes: Tubulin Tyrosine Ligase (TTL) and Tubulin Tyrosine Carboxypeptidase (TTCP). In this paper, I present evidence suggesting that the cycle itself is under the hierarchical control of reversible phosphorylation and that PKC mediated phosphorylation of TTL inhibits its activity, thereby preventing tubulin tyrosination. Phosphorylation of TTL is predicted to occur in a postulated Mg(++)/-ATP binding fold, leading to inhibition of Mg(++)/ATP binding and TTL mediated catalysis. The implications of such control are also discussed. PMID- 10842327 TI - Human papillomavirus type 16 E6 and HPV-16 E6/E7 sensitize human keratinocytes to apoptosis induced by chemotherapeutic agents: roles of p53 and caspase activation. AB - We and others have previously reported that human papillomavirus (HPV)-16 E6 protein expression sensitizes certain cell types to apoptosis. To confirm that this sensitization occurred in HPV's natural host cells, and to explore the mechanism(s) of sensitization, we infected human keratinocytes (HKCs) with retroviruses containing HPV-6 E6, HPV-16 E6, HPV-16 E7, or HPV-16 E6/E7. Apoptosis was monitored by DNA fragmentation gel analysis and direct observation of nuclei in cells stained with DAPI. Exposure of HKCs to etoposide, cisplatin, mitomycin C (MMC), atractyloside, and sodium butyrate, resulted in a time and dose-dependent induction of apoptosis. Expression of HPV-16 E6 and HPV-16 E6/E7, but not HPV-6 E6 or HPV-16 E7, enhanced the sensitivity of HKCs to cisplatin-, etoposide- and MMC-, but not atractyloside- or sodium butyrate-induced apoptosis. Expression of both HPV-16 E6 and HPV-16 E6/E7 decreased, but did not abolish, p53 protein levels relative to normal HKCs, and resulted in cytoplasmic localization of wt p53. P53 induction occurred in HPV-16 E6 and HPV-16 E6/E7 expressing cells after exposure to cisplatin or MMC, though never to levels found in normal untreated HKCs. P21 levels were decreased in HPV-16 E6 and HPV-16 E6/E7 expressing HKCs, and no induction of p21 was seen in these cells following exposure to cisplatin or MMC. Caspase-3 activity was found to be elevated in HPV 16 E6-expressing HKCs following exposure to cisplatin and MMC as documented by fluorometric and Western Blot analysis. Expression of wt CrmA or treatment of HPV 16 E6 expressing HKCs with the caspase-3 inhibitor DEVD.fmk prevented HPV-16 E6 induced sensitization in HKCs. These results suggest that HPV-16 E6 and HPV-16 E6/E7 expression sensitizes HKCs to apoptosis caused by cisplatin, etoposide and MMC, but not atractyloside or sodium butyrate. The data also suggest that wt p53 and caspase-3 activity are required for HPV-16 E6 and HPV-16 E6/E7-induced sensitization of HKCs to DNA damaging agents. PMID- 10842330 TI - Reorganization of microtubules in the amitotically dividing macronucleus of tetrahymena. AB - We developed a modified immunofluorescence protocol that permitted visualization of microtubules inside the macronucleus of the ciliate Tetrahymena. Although the amitotically dividing macronucleus lacks a spindle, an elaborate system of microtubules is assembled inside the macronucleus and between the macronucleus and the cortex. Microtubules could not be detected inside the interphase macronuclei. The early stage of macronuclear division was associated with the assembly of short macronuclear microtubules that localized randomly. The intramacronuclear microtubules were subsequently organized in a radial manner. During elongation of the macronucleus, the distribution of macronuclear microtubules changed from radial to parallel. During constriction of the macronucleus, dense and tangled macronuclear microtubules were detected at the region of nuclear constriction. In the cytosol, microtubules were linking the macronucleus and cell cortex. During recovery after drug-induced depolymerization, microtubules reassembled at multiple foci inside the macronucleus in close proximity to the chromatin. We propose that these microtubules play roles in chromatin partitioning, macronuclear constriction, and positioning of the macronucleus in relation to the cell cortex. PMID- 10842329 TI - Low concentrations of fibrinogen increase cell migration speed on fibronectin/fibrinogen composite cables. AB - Optimal cell migration rate in a given direction (velocity) is a function of speed and directional persistence. Migration speed has been reported to be a function of adhesion strength such that optimal cell migration occurs where the cell is able to form enough stable attachments for good traction while allowing attachments at the trailing end to be broken during locomotion. This is particularly important in peripheral nerve regeneration where rapid Schwann cell recruitment across the injury site will lead to better functional recovery and reduced end organ atrophy. The aim here was to investigate the effects of changing adhesion properties of Fn materials by adding fibrinogen in order to design an optimal material for repair processes. Cell migration on Fn/Fg-cables increased with increasing content of %Fg to a peak cell migration velocity (Schwann cells) of 49 microm/h, at 50% Fg. Further increases in Fg content hindered cell migration. Vinculin-rich attachment plaques were reduced in a dose dependent manner as the content of %Fg was increased whilst cells at the optimum Fg proportion for cell migration were moderately well spread. These results support the idea that optimum cell migration rates occur at intermediate attachment conditions, in this case at 50% Fg. These results show that incorporation of Fg into Fn-based materials will enhance the speed of Schwann cell migration and this would be likely to improve peripheral nerve regeneration. Indeed, directionally aligned Fn-based materials can now be engineered to give optimal cell velocity during repair cell recruitment in a range of tissue repair or tissue engineering applications. PMID- 10842331 TI - Melatonin induction of filamentous structures in non-neuronal cells that is dependent on expression of the human mt1 melatonin receptor. AB - Melatonin has gained recent popularity as a treatment for insomnia and other sleep disorders; however, its cellular effects are unknown. We report the effects of melatonin on the cellular morphology of Chinese hamster ovary (CHO) cells transformed to express the human melatonin receptors, mt1 and MT2. Our results show that melatonin exerts a strong influence on cellular shape and cytoskeletal organization in a receptor-dependent and possibly subtype-selective manner. The cell shape change that we see after a 5-h treatment of these non-neuronal cells with a pharmacological concentration of melatonin consists of the formation of long filamentous outgrowths that are reminiscent of the neurite processes produced by differentiating nerve cells. This morphological change occurs exclusively in cells expressing the mt1 receptor. We find that the microtubule and microfilament organization within these outgrowths is similar to that of neurites. Microtubules are required for the shape change to occur as Colcemid added in combination with melatonin completely blocks outgrowth formation. We demonstrate that the number of cells showing the altered cell shape is dependent on melatonin concentration, constant exposure to melatonin and that outgrowth frequencies increase when protein kinase A (PKA) is inhibited. Concomitant melatonin-dependent increases in MEK 1/2 and ERK 1/2 phosphorylation are noted in mt1-CHO cells only. The production of filamentous outgrowths is dependent on the translation of new protein but not the transcription of new mRNA. Outgrowth number is not controlled by centrosomes but is instead controlled by the polymerization state of the actin cytoskeleton. The results of this work show that the organization of the cytoskeleton is affected by processes specifically mediated or regulated by the mt1 receptor and may represent a novel alternative mechanism for the stimulation of process formation. PMID- 10842332 TI - Involvement of an F-actin skeleton on the acrosome reaction in guinea pig spermatozoa. AB - The acrosome reaction (AR) is a regulated exocytotic process. In several cell types, an actin network situated under the plasma membrane (PM) acts as a physical barrier to prevent this exocytosis. In seeking a function for a cortical skeleton in guinea pig spermatozoa, the PM and the outer acrosomal membrane (OAM) were investigated for the presence of F-actin and spectrin, proteins generally found in cell cortical skeletons. Both membrane types were visualized in whole mount preparations by electron microscopy. PM proteins gave positive reaction to the Na(+),K(+)-ATPase antibody and the OAM proteins did not react to the antibody. Furthermore, a Triton X-100-resistant skeleton was obtained from both membrane types. Using gold immunoelectron microscopy, F-actin was visualized in the PM and in the OAM skeletons, while spectrin was only detected in the PM skeleton. The presence of an F-actin cortical skeleton in the sperm PM suggests that F-actin may be involved in the AR. The significantly higher number of AR elicited by cytochalasin D (Cyt-D) treatment(P<0.005) and data showing a significant (P>0.03) decrease in F-actin relative concentration in capacitating spermatozoa, agree with this suggestion. Furthermore, the proposal is strengthened by the fact that stabilization of F-actin by phalloidin (Ph) significantly (P>0.01) diminished AR induced by Ca(2+) in a streptolysin O (SLO) permeabilized sperm model. PMID- 10842333 TI - Nonmuscle myosin II localizes to the Z-lines and intercalated discs of cardiac muscle and to the Z-lines of skeletal muscle. AB - To understand the role of nonmuscle myosin II in cardiac and skeletal muscle, we used a number of polyclonal antibodies, three detecting nonmuscle myosin heavy chain II-B (NMHC II-B) and two detecting NMHC II-A, to examine the localization of these two proteins in fresh-frozen, acetone-fixed sections of normal human and mouse hearts and human skeletal muscles. Results were similar in both species and were confirmed by examination of fresh-frozen sections of human hearts subjected to no fixation or to treatment with either 4% p-formaldehyde or 50% glycerol. NMHC II-B was diffusely distributed in the cytoplasm of cardiac myocytes during development, but after birth it was localized to the Z-lines and intercalated discs. Dual labeling showed almost complete colocalization of NMHC II-B with alpha-actinin. Whereas endothelial cells, smooth muscle cells and fibroblasts showed strong immunoreactivity for NMHC II-A and NMHC II-B, cardiac myocytes only showed reactivity for the latter. The Z-lines of human skeletal muscle cells, in contrast to those of cardiac myocytes, gave positive reactions for both NMHC II-A and NMHC II-B. The presence of a motor protein in the Z-lines and intercalated discs raises the possibility that these structures may play a more dynamic role in the contraction/relaxation mechanism of cardiac and skeletal muscle than has been previously suspected. PMID- 10842334 TI - Ion-induced conformational and stability changes in Nereis sarcoplasmic calcium binding protein: evidence that the APO state is a molten globule. AB - Nereis sarcoplasmic Ca(2+)-binding protein (NSCP) is a calcium buffer protein that binds Ca(2+) ions with high affinity but is also able to bind Mg(2+) ions with high positive cooperativity. We investigated the conformational and stability changes induced by the two metal ions. The thermal reversible unfolding, monitored by circular dichroism spectroscopy, shows that the thermal stability is maximum at neutral pH and increases in the order apo < Mg(2+) < Ca(2+). The stability against chemical denaturation (urea, guanidinium chloride) studied by circular dichroism or intrinsic fluorescence was found to have a similar ion dependence. To explore in more detail the structural basis of stability, we used the fluorescent probes to evaluate the hydrophobic surface exposure in the different ligation states. The apo-NSCP exhibits accessible hydrophobic surfaces, able to bind fluorescent probes, in clear contrast with denatured or Ca(2+)/Mg(2+)-bound states. Gel filtration experiments showed that, although the metal-bound NSCP has a hydrodynamic volume in agreement with the molecular mass, the volume of the apo form is considerably larger. The present results demonstrate that the apo state has many properties in common with the molten globule. The possible factors of the metal-dependent structural changes and stability are discussed. PMID- 10842335 TI - Empirical calculation of the relative free energies of peptide binding to the molecular chaperone DnaK. AB - We describe a methodology to calculate the relative free energies of protein peptide complex formation. The interaction energy was decomposed into nonpolar, electrostatic and entropic contributions. A free energy-surface area relationship served to calculate the nonpolar free energy term. The electrostatic free energy was calculated with the finite difference Poisson-Boltzmann method and the entropic contribution was estimated from the loss in the conformational entropy of the peptide side chains. We applied this methodology to a series of DnaK*peptide complexes. On the basis of the single known crystal structure of the peptide-binding domain of DnaK with a bound heptapeptide, we modeled ten other DnaK*heptapeptide complexes with experimentally measured K(d) values from 0.06 microM to 11 microM, using molecular dynamics to refine the structures of the complexes. Molecular dynamic trajectories, after equilibration, were used for calculating the energies with greater accuracy. The calculated relative binding free energies were compared with the experimentally determined free energies. Linear scaling of the calculated terms was applied to fit them to the experimental values. The calculated binding free energies were between -7.1 kcal/mol and - 9.4 kcal/mol with a correlation coefficient of 0.86. The calculated nonpolar contributions are mainly due to the central hydrophobic binding pocket of DnaK for three amino acid residues. Negative electrostatic fields generated by the protein increase the binding affinity for basic residues flanking the hydrophobic core of the peptide ligand. Analysis of the individual energy contributions indicated that the nonpolar contributions are predominant compared to the other energy terms even for peptides with low affinity and that inclusion of the change in conformational entropy of the peptide side chains does not improve the discriminative power of the calculation. The method seems to be useful for predicting relative binding energies of peptide ligands of DnaK and might be applicable to other protein-peptide systems, particularly if only the structure of one protein-ligand complex is available. PMID- 10842336 TI - Solvent density and long-range dipole field around a DNA-binding protein studied by molecular dynamics. AB - The distribution and orientation of solvent around a DNA-binding protein, 434 Cro, were investigated by molecular dynamics simulations with a periodic-boundary condition. The protein was treated in two states: charged and neutral. The computed high-density sites of the solvent around the protein correlated well with the experimentally determined crystal-water sites, in both the charged and neutral states. A local density map, introduced to investigate the solvent density around the highly mobile regions of the protein, showed a hydration shell around hydrophobic sidechains and hydrogen-bondable sites around hydrophilic sidechains, and also showed that the solvent density is sensitive to the slight concaves of the sidechain surface. The long-range solvent-dipole field was observed around the protein, where the pattern of the dipole ordering was considerably different between the charged and neutral states. A local solvent dipole field was introduced, and the pattern of the dipole ordering was different between the hydrophobic and hydrophilic sidechains. The dipole field from the charged state provided a higher correlation to the electrostatic field obtained from the Poisson-Boltzmann's equation than that from the neutral state, although the correlation become weak quickly for the both states with increasing the protein-solvent distance. PMID- 10842337 TI - Electrostatics of mesophilic and psychrophilic trypsin isoenzymes: qualitative evaluation of electrostatic differences at the substrate binding site. AB - A qualitative evaluation of electrostatic features of the substrate binding region of seven isoenzymes of trypsin has been performed by using the continuum electrostatic model for the solution of the Poisson-Boltzmann equation. The sources of the electrostatic differences among the trypsins have been sought by comparative calculations on selective charges: all charges, conserved charges, partial charges, unique cold trypsin charges, and a number of charge mutations. As expected, most of the negative potential at the S(1) region of all trypsins is generated from Asp(189), but the potential varies significantly among the seven trypsin isoenzymes. The three cold active enzymes included in this study possess a notably lower potential at and around the S(1)-pocket compared with the warm active counterparts; this finding may be the main contribution to the increased binding affinity. The source of the differences are nonconserved charged residues outside the specificity pocket, producing electric fields at the S(1)-pocket that are different in both sign and magnitude. The surface charges of the mesophilic trypsins generally induce the S(1) pocket positively, whereas surface charges of the cold trypsins produce a negative electric field of this region. Calculations on mutants, where charged amino acids were substituted between the trypsins, showed that mutations in Loop2 (residues 221B and 224) and residue 175, in particular, were responsible for the low potential of the cold enzymes. PMID- 10842338 TI - NMR investigations of protein-carbohydrate interactions: studies on the relevance of Trp/Tyr variations in lectin binding sites as deduced from titration microcalorimetry and NMR studies on hevein domains. Determination of the NMR structure of the complex between pseudohevein and N,N',N"-triacetylchitotriose. AB - Model studies on lectins and their interactions with carbohydrate ligands in solution are essential to gain insights into the driving forces for complex formation and to optimize programs for computer simulations. The specific interaction of pseudohevein with N,N', N"-triacetylchitotriose has been analyzed by (1)H-NMR spectroscopy. Because of its small size, with a chain length of 45 amino acids, this lectin is a prime target to solution-structure determination by NOESY NMR experiments in water. The NMR-analysis was extended to assessment of the topology of the complex between pseudohevein and N, N',N" triacetylchitotriose. NOESY experiments in water solution provided 342 protein proton-proton distance constraints. Binding of the ligand did not affect the pattern of the protein nuclear Overhauser effect signal noticeably, what would otherwise be indicative of a ligand-induced conformational change. The average backbone (residues 3-41) RMSD of the 20 refined structures was 1.14 A, whereas the heavy atom RMSD was 2.18 A. Two different orientations of the trisaccharide within the pseudohevein binding site are suggested, furnishing an explanation in structural terms for the lectin's capacity to target chitin. In both cases, hydrogen bonds and van der Waals contacts confer stability to the complexes. This conclusion is corroborated by the thermodynamic parameters of binding determined by NMR and isothermal titration calorimetry. The association process was enthalpically driven. In relation to hevein, the Trp/Tyr-substitution in the binding pocket has only a small effect on the free energy of binding in contrast to engineered galectin-1 and a mammalian C-type lectin. A comparison of the three dimensional structure of pseudohevein in solution to those reported for wheat germ agglutinin (WGA) in the solid state and for hevein and WGA-B in solution has been performed, providing a data source about structural variability of the hevein domains. The experimentally derived structures and the values of the solvent accessibilities for several key residues have also been compared with conformations obtained by molecular dynamics simulations, pointing to the necessity to further refine the programs to enhance their predictive reliability and, thus, underscoring the importance of this kind of combined analysis in model systems. PMID- 10842339 TI - Toward an energy function for the contact map representation of proteins. AB - We analyzed several energy functions for predicting the native state of proteins from an energy minimization procedure. We derived the parameters of a given energy function by imposing the basic requirement that the energy of the native conformation of a protein is lower than that of any conformation chosen from a set of decoys. Our work is motivated by a recent result which proved that the simple pairwise contact approximation of the energy is insufficient to satisfy simultaneously such a basic requirement for all the proteins in a database. Here, we investigate the reasons of such negative results and show how to improve the predictive power of methods based on energy minimization. We generated decoys by gapless threading, and we derive energy parameters by perceptron learning. We first considered hydrophobic contributions to the energy, defined in several ways, and showed that the additional hydrophobic terms enlarge slightly the number of proteins that can be stabilized together. Next, we performed various modifications of the pairwise energy term. We introduced (1) a distinction between inter-residue contacts on the surface and in the core of a protein and (2) a simple distance-dependent pairwise interaction in which a two-tier definition of contact replaces the original (single-tier) one. Our results suggest that a detailed treatment of the pairwise potential is likely to be more relevant than the consideration of other forces. PMID- 10842340 TI - Crystal structure of recombinant trypsin-solubilized fragment of cytochrome b(5) and the structural comparison with Val61His mutant. AB - The crystal structure of the recombinant trypsin-solubilized fragment of the microsomal cytochrome b(5) from bovine liver has been determined at 1.9 A resolution and compared with the reported crystal structure of the lipase solubilized fragment of the membrane protein cytochrome b(5). The two structures are similar to each other. However, some detailed structural differences are observed: the conformation of the segment Asn16-Ser20 is quite different, some helices around the heme and some segments between the helices are shifted slightly, the heme is rotated about the normal of the mean plane of heme, one of the propionates of the heme exhibits a different conformation. The average coordination distances between the iron and the two nitrogen atoms of the imidazole ligands are the same in the two structures. Most of the structural differences can be attributed to the different intermolecular interactions which result from the crystal packing. The wild-type protein structure is also compared with its Val61His mutant, showing that the heme binding and the main chain conformations are basically identical with each other except for the local area of the mutation site. However, when Val61 is mutated to histidine, the large side chain of His61 is forced to point away from the heme pocket toward the solvent region, disturbing the micro-environment of the heme pocket and influencing the stability and the redox potential of the protein. PMID- 10842341 TI - Dynamic ligand design and combinatorial optimization: designing inhibitors to endothiapepsin. AB - The dynamic ligand design (DLD) algorithm, an automated method for the creation of novel ligands, links up small functional groups that have been placed in energetically favorable positions in the binding site of a target molecule. The positions and orientations of the small functional groups can be determined using the multi-copy simultaneous search approach (MCSS) or experimental data. In this work the original DLD methodology is extended by using a modified version of the pseudo-potential energy function. A novel simulated annealing protocol is presented for optimizing the pseudo-potential energy of ligands in the binding site; the protocol is expected to be applicable to other optimization problems. The utility of the method is illustrated by designing an inhibitor for endothiapepsin. The binding affinity of the inhibitor is assessed using a thermodynamic cycle that decomposes the binding free energy into a sum of translational, rotational, configurational, hydrophobic, and electrostatic contributions. The calculations suggest that the designed molecule will bind endothiapepsin with high affinity. PMID- 10842342 TI - Comparative X-ray analysis of the un-liganded fosfomycin-target murA. AB - MurA, an essential enzyme for the synthesis of the bacterial cell wall, follows an induced-fit mechanism. Upon substrate binding, the active site forms in the interdomain cleft, involving movements of the two domains of the protein and a reorientation of the loop Pro112-Pro121. We compare two structures of un-liganded MurA from Enterobacter cloacae: a new orthorhombic form, solved to 1.80 A resolution, and a monoclinic form, redetermined to 1.55 A resolution. In the monoclinic form, the loop Pro112-Pro121 stretches into solvent, while in the new form it adopts a winded conformation, thereby reducing solvent accessibility of the critical residue Cys115. In the interdomain cleft a network of 27 common water molecules has been identified, which partially shields negative charges in the cleft and stabilizes the orientation of catalytically crucial residues. This could support substrate binding and ease domain movements. Near the hinge region an isoaspartyl residue has been recognized, which is the product of post translational modification of the genetically encoded Asn67-Gly68. The homogeneous population with L-isoaspartate in both structures suggests that the modification in Enterobacter cloacae MurA is not a mere aging defect but rather the result of a specific in vivo process. PMID- 10842343 TI - Automated docking of alpha-(1-->4)- and alpha-(1-->6)-linked glucosyl trisaccharides and maltopentaose into the soybean beta-amylase active site. AB - The Lamarckian genetic algorithm of AutoDock 3.0 was used to dock alpha maltotriose, methyl alpha-panoside, methyl alpha-isopanoside, methyl alpha isomaltotrioside, methyl alpha-(6(1)-alpha-glucopyranosyl)-maltoside, and alpha maltopentaose into the closed and, except for alpha-maltopentaose, into the open conformation of the soybean beta-amylase active site. In the closed conformation, the hinged flap at the mouth of the active site closes over the substrate. The nonreducing end of alpha-maltotriose docks preferentially to subsites -2 or +1, the latter yielding nonproductive binding. Some ligands dock into less optimal conformations with the nonreducing end at subsite -1. The reducing-end glucosyl residue of nonproductively-bound alpha-maltotriose is close to residue Gln194, which likely contributes to binding to subsite +3. In the open conformation, the substrate hydrogen-bonds with several residues of the open flap. When the flap closes, the substrate productively docks if the nonreducing end is near subsites 2 or -1. Trisaccharides with alpha-(1-->6) bonds do not successfully dock except for methyl alpha-isopanoside, whose first and second glucosyl rings dock exceptionally well into subsites -2 and -1. The alpha-(1-->6) bond between the second and third glucosyl units causes the latter to be improperly positioned into subsite +1; the fact that isopanose is not a substrate of beta-amylase indicates that binding to this subsite is critical for hydrolysis. PMID- 10842344 TI - An atoms-in-molecules study of the genetically-encoded amino acids: I. Effects of conformation and of tautomerization on geometric, atomic, and bond properties. AB - The theory of Atoms-In-Molecules (AIM) is a partitioning of the real space of a molecule into disjoint atomic constituents as determined by the topology of the electron density, rho(r). This theory identifies an atom in a molecule with a quantum mechanical open system and, consequently, all of the atom's properties are unambiguously defined. AIM recovers the basic empirical cornerstone of chemistry: that atoms and functional groups possess characteristic and additive properties that in many cases exhibit a remarkable transferability between different molecules. As a result, the theory enables the theoretical synthesis of a large molecule and the prediction of its properties by joining fragments that are predetermined as open systems. The present article is the first of a series (in preparation) that explore this possibility for polypeptides by determining the transferability of the building blocks: the amino acid residues. Transferability of group properties requires transferability of the electron density rho(r), which in turn requires the transferability of the geometric parameters. This article demonstrates that these parameters are conformation insensitive for a representative amino acid, leucine, and that the atomic and bond properties exhibit a corresponding transferability. The effects of hydrogen bonding are determined and a set of geometrical conditions for the occurrence of such bonding is identified. The effects of transforming neutral leucine into its zwitter-ionic form on its atomic and bond properties are shown to be localized primarily to the sites of ionization. PMID- 10842345 TI - Key residues approach to the definition of protein families and analysis of sparse family signatures. AB - We extend the concept of the motif as a tool for characterizing protein families and explore the feasibility of a sparse "motif" that is the length of the protein sequence itself. The type of motif discussed is a sparse family signature consisting of a set of N key residue positions (A1, A2...AN) preceded by gaps (G) thus G1A1G2A2. ...GNAN. Both a residue and gap can be variable. A signature is matched to a protein sequence and scored using a dynamic programming algorithm which permits variability in gap distance and residue type. Generating a signature involves identifying residues associated with points of contact in interactions between secondary structure elements. A raw signature consists of a set of positions with potential key structural roles sampled from a sequence alignment constructed with reference to this contact data. Raw signatures are refined by sampling different gap-residue pairs until the specificity of a signature for the family cannot be further improved. We summarize signatures for nine families of protein of diverse fold and function and present results of scans against the OWL protein sequence database. The implications of such signatures are discussed. PMID- 10842346 TI - Dopamine D(1) and D(2) receptors in the forebrain of dystonia musculorum mutant mice: an autoradiographic survey in relation to dopamine contents. AB - Dystonia musculorum (dt(J)/dt(J)) mutant mice suffer from a degeneration of spinocerebellar tracts as well as a dystrophy of peripheral sensory tracts. This neurological mutant has been proposed as an animal model of human cerebellar ataxia, in particular of the Friedreich's type; thus, it was deemed of interest to examine the endogenous contents of dopamine (DA) and metabolites as well as the distribution of DA receptors of the D(1) and D(2) subtypes, in order to delimit the biochemical characteristics of this pathological disorder, and determine an eventual dopaminergic dysfunction in this mutant. Tissue DA and its major metabolites 3, 4-dihydroxyphenylacetic acid, homovanillic acid and 3 methoxytyramine were measured by HPLC coupled to electrochemical detection in six cortical regions, in four divisions of rostral neostriatum and two halves of caudal neostriatum, as well as in olfactory bulb, nucleus accumbens, septum, amygdala, hippocampus, thalamus, hypothalamus, brainstem, cerebellum, substantia nigra, and ventral tegmental area. The only significant difference between dt(J)/dt(J) mice and wild-type controls was an increase in hypothalamic DA contents (+47%). Quantitative autoradiography with [(3)H]SCH23390 and [(3)H]raclopride, to label D(1) and D(2) receptors, respectively, revealed only moderate changes in receptor densities in a few localized regions. In dt(J)/dt(J) mutants, D(1) receptor numbers were found to be higher in thalamus (+27%) as well as in the medio-dorsal (+16%) and in the latero-dorsal (+16%) quadrants of rostral neostriatum, while D(2) receptor densities were greater in the medio ventral (+32%) and the latero-dorsal (+17%) quadrants. The present results indicate an overall conservation of dopaminergic functions, albeit the few localized sites of increased D(1) and D(2) receptor densities, and that are seemingly independent of the DA innervation pattern, as revealed by the tissue measurements of DA and metabolites. They also rule out a major pathology linked to deficits in DA neurotransmission, and validate this mutant as an animal model of human cerebellar ataxia, probably of the Friedreich type. PMID- 10842347 TI - Morphology of layer III pyramidal neurons is altered following induction of LTP in sensorimotor cortex of the freely moving rat. AB - The organization of specific cortical connections can be altered by sensory and motor experience. These changes are believed to result from activity-dependent changes in synaptic connectivity, similar to those induced in the hippocampus by high-frequency stimulation in long-term potentiation (LTP) experiments. If similar mechanisms are involved, then neocortical LTP induction may induce some of the same morphological changes that are seen following learning. We induced LTP in the contralateral sensorimotor cortex by repeated, daily tetanization of the corpus callosum in chronically implanted, freely moving rats. Anatomical results showed that the LTP induction was associated with alterations in dendrite morphology and increased spine density. These changes are qualitatively and quantitatively similar to those commonly observed in studies in which rats are housed in complex environments. The similarity of results following exposure to complex environments and after LTP induction in the neocortex may indicate a reliance on the same cellular mechanisms in both situations. PMID- 10842348 TI - Prenatal protein malnutrition results in increased frequency of miniature inhibitory synaptic currents in rat CA1 pyramidal cells. AB - There is growing evidence for an effect of prenatal protein malnutrition on the GABAergic neurotransmitter system in the rat hippocampus and associated structures. In the present study, we examined the functional electrophysiological consequences of observed alterations in GABA(A) and benzodiazepine receptor systems. Whole-cell patch clamp recordings of spontaneous and of miniature inhibitory postsynaptic currents (mIPSCs) generated by CA1 pyramidal cells were performed in in vitro hippocampal slices prepared from control and prenatally protein malnourished adult male rats. The characteristics of spontaneous synaptic currents were unaltered by the prenatal insult, as were the amplitudes and kinetics of GABA(A) receptor-mediated mIPSCs. The frequency of mIPSCs, however, was significantly increased in CA1 pyramidal cells in slices prepared from prenatally malnourished vs. control rats. The effect of the benzodiazepine receptor agonist chlordiazepoxide on the characteristics of mIPSCs was also examined and found to be the same in cells from both nutritional groups. The increased frequency of mIPSCs together with the lack of a change in amplitude, kinetics, or modulation by benzodiazepines of mIPSCs in response to prenatal protein malnutrition indicate a presynaptic locus of effect of this insult. PMID- 10842349 TI - Repeated administration of methamphetamine damages cells in the somatosensory cortex: overlap with cytochrome oxidase-rich barrels. AB - Multiple injections of methamphetamine (mAMPH) cause degeneration of neurons in rat primary somatosensory cortex (Par1). These degenerating cells can be labeled histochemically with the fluorochrome dye, Fluoro-Jade (FJ). This area of Par1 also contains the representation of the mystacial vibrissae. Neurons in this area of Par1 receiving projections derived from the vibrissae are arranged in discrete functional units ("barrels"), which are revealed by cytochrome oxidase (CO) histochemistry. Here, rats given mAMPH (four injections of 4 mg/kg, sc, 2-h intervals between injections) showed FJ-positive neurons in Par1 that were located predominantly near the perimeter of the CO-dense barrels. Thus, the Par1 neurons damaged by multiple administration of mAMPH are located within whisker barrels. PMID- 10842350 TI - Surface-accessible GABA supports tonic and quantal synaptic transmission. AB - Exocytosis is commonly viewed as the only secretory process able to account for quantal forms of fast synaptic transmission. However, the demonstrated variability and composite properties of miniature postsynaptic signals are not easily explained by all-or-none exocytotic discharge of transmitter in solution from inside vesicles. Recent studies of endocrine secretion have shown that hormone release does not coincide with exocytosis due to its trapping in the core matrix of the granule. Thus, we tested whether the synaptic transmitter GABA could also be held in a matrix before being released. Using confocal microscopy and flow cytometry of embryonic rat hippocampal neurons, we found a GABA immunoreaction at the surface of live cell bodies and growth cones that coincided spatially and quantitatively with the binding of tetanus toxin fragment C (TTFC). TTFC binds predominantly at membrane sites containing the trisialoglycosphingolipid GT1b. Using flow cytometry, GT1b-containing liposomes preincubated in 100 nM GABA exhibited the same relationship between GABA and TTFC surface binding as found on neurons and growth cones. Embryonic neurons differentiated in culture expressed initially a tonic, and after 3-5 days, transient, postsynaptic signals mediated by GABA acting at GABA(A) receptor/Cl(-) channels. A stream of saline applied to the neuronal surface rapidly and reversibly suppressed both tonic and transient signals. A brief application of the GABAmimetic isoguvacine immediately transformed both tonic and transient GABAergic signals into tonic and transient isoguvacinergic signals. These results and those in the literature are consistent with an immediately releasable compartment of transmitter accessible from the presynaptic surface. PMID- 10842352 TI - In vitro and in vivo evaluation of the binding of the dopamine D2 receptor agonist (11)C-(R,S)-5-hydroxy-2-(di-n-propylamino)tetralin in rodents and nonhuman primate. AB - The in vitro autoradiographic binding characteristics as well as in vivo imaging characteristics of a dopamine D2 receptor agonist, (R, S)-2-(N-propyl-N-1'-(11)C propyl)amino-5-hydroxytetralin ((11)C-5-OH-DPAT), were studied. In (3)H-spiperone assays using rat striata, 5-OH-DPAT exhibited an affinity of IC(50) = 2.5 nM. In vitro autoradiographs in rat brain slices with (11)C-5-OH-DPAT revealed selective binding to the dopaminergic regions in the striata which was displaceable by sulpiride. Varying concentrations of dopamine displaced this selective binding of (11)C-5-OH-DPAT to the striata in rat brain slices. This selective binding to the striata was also removed in the presence of the GTP analog, 5' guanylylimidodiphosphate, indicative of the binding of (11)C-5-OH-DPAT to the high-affinity state of the D2 receptor. Ex vivo autoradiographic study in rats exhibited selective binding of (11)C-5-OH-DPAT to the striata. A PET study in a rhesus monkey showed selective localization of (11)C-5-OH-DPAT in the striata and the ratio between striata and cerebellum approached approximately 2 at 40 min postinjection. PMID- 10842351 TI - Enhancement of NMDA-induced current by the putative NR2B selective antagonist ifenprodil. AB - Ifenprodil has been widely used as an antagonist selective for NMDA receptors containing the NR2B subunit. Evidence suggests, however, that ifenprodil also increases NMDA receptor affinity. Using rat brain slices, we found that ifenprodil enhanced NMDA-induced current in both cortical and subcortical areas examined. To test whether the effect is due to an increase in NMDA receptor affinity, we compared the effect of ifenprodil on currents induced by different concentrations of NMDA. Consistent with the hypothesis, the enhancing effect (percent increase) was relatively constant at low NMDA concentrations. As NMDA concentration increased, however, the effect decreased. To test whether the effect is blocked when NMDA binding sites are saturated with NMDA, high concentrations of NMDA were applied. To partially block Ca(2+) influx and prevent cells from deteriorating, the experiments were performed in the presence of either MK801 or kynurenate, two noncompetitive antagonists. Under such conditions, ifenprodil not only failed to potentiate NMDA currents, but consistently suppressed the current. When the same concentration of NMDA was applied in the presence of the competitive antagonist CGP37849, ifenprodil regained its ability to potentiate NMDA currents. Furthermore, the higher the concentration of CGP37849 the more the NMDA current was potentiated by ifenprodil. These results, combined with previous studies, suggest that the enhancing effect is due to an increase in NMDA receptor affinity and is specific for responses induced by low NMDA concentrations. As NMDA concentration increases, the affinity-enhancing effect decreases. Consequently, the channel suppressing effect becomes more prominent. PMID- 10842353 TI - Localization of cannabinoid CB(1) receptor mRNA in neuronal subpopulations of rat striatum: a double-label in situ hybridization study. AB - Double-label in situ hybridization was used to identify the phenotypes of striatal neurons that express mRNA for cannabinoid CB(1) receptors. Simultaneous detection of multiple mRNAs was performed by combining a (35)S-labeled ribonucleotide probe for CB(1) mRNA with digoxigenin-labeled riboprobes for striatal projection neurons (preprotachykinin A, prodynorphin, and preproenkephalin mRNAs) and interneurons (vesicular acetylcholine transporter (VAChT), choline acetyltransferase (ChAT), somatostatin, and glutamic acid decarboxylase (Mr 67,000; GAD67) mRNAs). To ascertain whether CB(1) mRNA was a marker for striatal efferents, digoxigenin-labeled probes for mRNA markers of both striatonigral (prodynorphin or preprotachykinin A mRNAs), and striatopallidal (proenkephalin mRNAs) projection neurons were combined with the (35)S-labeled probe for CB(1). A mediolateral gradient in CB(1) mRNA expression was observed at rostral and mid-striatal levels; in the same coronal sections the number of silver grains per cell ranged from below the threshold of detectability at the medial and ventral poles to saturation at the dorsolateral boundary bordered by the corpus callosum. At the caudal level examined, CB(1) mRNA was denser in the ventral sector relative to the dorsal sector. Virtually all neurons expressing mRNA markers for striatal projection neurons colocalized CB(1) mRNA. Combining a (35)S-labeled riboprobe for CB(1) with digoxigenin-labeled riboprobes for both preproenkephalin and prodynorphin confirmed localization of CB(1) mRNA to striatonigral and striatopallidal neurons expressing prodynorphin and preproenkephalin mRNAs, respectively. However, CB(1) mRNA-positive cells that failed to coexpress the other markers were also apparent. CB(1) mRNA was localized to putative GABAergic interneurons that express high levels of GAD67 mRNA. These interneurons enable functional interactions between the direct and indirect striatal output pathways. By contrast, aspiny interneurons that express preprosomatostatin mRNA and cholinergic interneurons that coexpress ChAT and VAChT mRNAs were CB(1) mRNA-negative. The present data provide direct evidence that cannabinoid receptors are synthesized in striatonigral neurons that contain dynorphin and substance P and striatopallidal neurons that contain enkephalin. By contrast, local circuit neurons in striatum that contain somatostatin or acetylcholine do not synthesize cannabinoid receptors. Published 2000 Wiley-Liss, Inc. PMID- 10842354 TI - Form and function: the laminin family of heterotrimers. AB - The laminins are a family of glycoproteins that provide an integral part of the structural scaffolding of basement membranes in almost every animal tissue. Each laminin is a heterotrimer assembled from alpha, beta, and gamma chain subunits, secreted and incorporated into cell-associated extracellular matrices. The laminins can self-assemble, bind to other matrix macromolecules, and have unique and shared cell interactions mediated by integrins, dystroglycan, and other receptors. Through these interactions, laminins critically contribute to cell differentiation, cell shape and movement, maintenance of tissue phenotypes, and promotion of tissue survival. Recent advances in the characterization of genetic disruptions in humans, mice, nematodes and flies have revealed developmental roles for the different laminin subunits in diverse cell types, affecting differentiation from blastocyst formation to the post-natal period. These genetic defects have challenged some of the previous concepts about basement membranes and have shed new light on the diversity and complexity of laminin functions as well as established the molecular basis of several human diseases. PMID- 10842355 TI - The tenascin family of ECM glycoproteins: structure, function, and regulation during embryonic development and tissue remodeling. AB - The determination of animal form depends on the coordination of events that lead to the morphological patterning of cells. This epigenetic view of development suggests that embryonic structures arise as a consequence of environmental influences acting on the properties of cells, rather than an unfolding of a completely genetically specified and preexisting invisible pattern. Specialized cells of developing multicellular organisms are surrounded by a complex extracellular matrix (ECM), comprised largely of different collagens, proteoglycans, and glycoproteins. This ECM is a substrate for tissue morphogenesis, lends support and flexibility to mature tissues, and acts as an epigenetic informational entity in the sense that it transduces and integrates intracellular signals via distinct cell surface receptors. Consequently, ECM receptor interactions have a profound influence on major cellular programs including growth, differentiation, migration, and survival. In contrast to many other ECM proteins, the tenascin (TN) family of glycoproteins (TN-C, TN-R, TN-W, TN-X, and TN-Y) display highly restricted and dynamic patterns of expression in the embryo, particularly during neural development, skeletogenesis, and vasculogenesis. These molecules are reexpressed in the adult during normal processes such as wound healing, nerve regeneration, and tissue involution, and in pathological states including vascular disease, tumorigenesis, and metastasis. In concert with a multitude of associated ECM proteins and cell surface receptors that include members of the integrin family, TN proteins impart contrary cellular functions, depending on their mode of presentation (i.e., soluble or substrate bound) and the cell types and differentiation states of the target tissues. Expression of tenascins is regulated by a variety of growth factors, cytokines, vasoactive peptides, ECM proteins, and biomechanical factors. The signals generated by these factors converge on particular combinations of cis-regulatory elements within the recently identified TN gene promoters via specific transcriptional activators or repressors. Additional complexity in regulating TN gene expression is achieved through alternative splicing, resulting in variants of TN polypeptides that exhibit different combinations of functional protein domains. In this review, we discuss some of the recent advances in TN biology that provide insights into the complex way in which the ECM is regulated and how it functions to regulate tissue morphogenesis and gene expression. PMID- 10842356 TI - Cellular signaling by neural cell adhesion molecules of the immunoglobulin superfamily. AB - Neural cell adhesion molecules (CAMs) of the immunoglobulin superfamily nucleate and maintain groups of cells at key sites during early development and in the adult. In addition to their adhesive properties, binding of CAMs can affect intracellular signaling. Their ability to influence developmental events, including cell migration, proliferation, and differentiation can therefore result both from their adhesive as well as their signaling properties. This review focuses on the two CAMs for which the most information is known, the neural CAM, N-CAM, and L1. N-CAM was the first CAM to be characterized and, therefore, has been studied extensively. The binding of N-CAM to cells leads to a number of signaling events, some of which result in changes in gene expression. Interest in L1 derives from the fact that mutations in its gene lead to human genetic diseases including mental retardation. Much is known about modifications of the L1 cytoplasmic domain and its interaction with cytoskeletal molecules. The study of CAM signaling mechanisms has been assay-dependent rather than molecule dependent, with particular emphasis on assays of neurite outgrowth and gene expression, an emphasis that is maintained throughout the review. The signals generated following CAM binding that lead to alterations in cell morphology and gene expression have been linked directly in only a few cases. We also review information on other CAMs, giving special consideration to those that are anchored in the membrane by a phospholipid anchor. These proteins, including a form of N-CAM, are presumed to be localized in lipid rafts, membrane substructures that include distinctive subsets of cytoplasmic signaling molecules such as members of the src-family of nonreceptor protein tyrosine kinases. In the end, these studies may reveal that what CAMs do after they bind cells together may have as profound consequences for the cells as the adhesive interactions themselves. This area will therefore remain a rich ground for future studies. PMID- 10842357 TI - The thrombospondin type 1 repeat (TSR) superfamily: diverse proteins with related roles in neuronal development. AB - The thrombospondins are a family of proteins found widely in the embryonic extracellular matrix. Like most matrix proteins, thrombospondins are modular and contain a series of repeated domains arrayed between globular amino and carboxyl terminal domains. In recent years, other proteins that share thrombospondin type 1 repeats, or TSRs, have been identified. These include the F-spondin gene family, the members of the semaphorin 5 family, UNC-5, SCO-spondin, and others. Most of these are expressed in the developing nervous system, and many have expression patterns and in vitro properties that suggest potential roles in the guidance of cell and growth cone migration. Both cell- and matrix-binding motifs have been identified in the TSRs of thrombospondin-1, so it has been hypothesized that the properties of these diverse proteins may also depend on the presence of these repeats. Here, we review the cell biology of the TSR module, the extensive literature regarding the distribution and functions of thrombospondins and other TSR superfamily proteins, and evaluate their possible roles during the development of the nervous system. PMID- 10842358 TI - Tubedown-1, a novel acetyltransferase associated with blood vessel development. AB - We have used an embryonic endothelial cell line (IEM cells) as an experimental system for identifying and characterizing new molecules which are regulated during blood vessel development. A novel gene isolated from IEM cells, tubedown-1 (tbdn-1), is expressed at high levels in unstimulated IEM cells and is downregulated during formation of capillary tube structures by the IEM cells induced by basic fibroblast growth factor (bFGF) and leukemia inhibitory factor (LIF) in vitro. Tbdn-1 is also downregulated in M1 myeloid leukemia cells after differentiation in response to LIF in vitro. Tbdn-1 is homologous to the yeast NAT-1 N-terminal acetyltransferases and encodes a novel protein of approximately 69 kDa associated with an acetyltransferase activity. Levels and distribution of tbdn-1 expression are regulated in both endothelial and hematopoietic cells during development in tissues such as the yolk sac blood islands, heart, and liver blood vessels. In the adult, tbdn-1 expression is low or undetected in most organs examined with the exception of the atrial endocardium, the endothelial and myeloid compartments of bone marrow, and the remodeling vascular bed of atretic ovarian follicles. The distribution and regulation of expression of tbdn-1 suggest that this novel acetyltransferase may be involved in regulating vascular and hematopoietic development and physiologic angiogenesis. PMID- 10842359 TI - Evidence that translation of smooth muscle alpha-actin mRNA is delayed in the chick promyocardium until fusion of the bilateral heart-forming regions. AB - Heart development in the chick embryo proceeds from bilateral mesodermal primordia established during gastrulation. These primordia migrate to the midline and fuse into a single heart trough. During their migration as a cohesive sheet, the cells of the paired heart fields become epithelial and undergo cardiac differentiation, exhibiting organized myofibrils and rhythmic contractions near the time of their fusion. Between the stages of cardiomyoblast commitment and overt differentiation of cardiomyocytes, a significant time interval exists. Using a new riboprobe (usmaar) for whole-mount in situ hybridization in chick embryos, we report the earliest phases of smooth muscle alpha-actin (smaa) mRNA distribution during the precontractile developmental window. We show that ingressed heart-forming regions express smaa by the head-process stage (Hamburger and Hamilton stage 5). In addition, we used usmaar to study the formation and early morphogenesis of the heart. Consistent with fate mapping studies (Garcia Martinez and Schoenwolf [1993] Dev. Biol. 159:706-719; Schoenwolf and Garcia Martinez [1995] Cell Mol. Biol. Res. 41:233-240; Garcia-Martinez et al., in preparation), our results with this probe, combined with detailed histological and SEM analyses of the so-called cardiac crescent, demonstrate unequivocally that the heart arises from separated and paired heart rudiments, rather than from a single crescent-shaped rudiment (that is, prior to fusion of the paired heart rudiments to establish the straight-heart tube, the rostral midline of the cardiac crescent lacks mesodermal cells and consequently fails to label with usmaar). Smaa is also expressed in the splanchnic and somatic mesoderm, marking the earliest step in coelom formation. Consequently, we also used usmaar to describe formation of the pericardium. Finally, we provide evidence of a post transcriptional level of control of smaa gene expression in the heart fields. Our results suggest that the expression of smaa may mark a primitive mesodermal state from which definitive cell types can be derived through inductive events. PMID- 10842360 TI - Connexin37 in normal and pathological development of mouse heart and great arteries. AB - Cx37 is a member of the connexin family of gap junction proteins, whose distribution in heart remains controversial. We have generated novel antibodies against Cx37 to investigate this distribution during normal and pathological development in mouse. Using these affinity-purified antibodies, we have detected Cx37 in hearts and aortas of mouse embryos from day 11 ed. onwards. Immunostaining revealed that during prenatal development Cx37 predominated in endothelial and endocardial cells but was also detectable in small amounts in the trabeculated and compact layers of ventricular myocardium, as well as in the mesenchyme of conotruncal ridges and atrioventricular cushions. Cx37 was also differentially expressed in the ascending and descending portions of the embryonic aorta, according to a pattern which differed in the three layers of the vessel wall. Cx37 distribution was altered in both heart and aorta of mice that had been exposed to all-trans retinoic acid at the beginning of foetal development, whether or not these animals subsequently developed a transposition of great arteries. The data indicate that Cx37 is widely distributed in multiple compartments of cardiovascular system, in patterns which are modulated during development, by retinoic acid. PMID- 10842361 TI - Combinatorial expression of zebrafish Brn-1- and Brn-2-related POU genes in the embryonic brain, pronephric primordium, and pharyngeal arches. AB - Vertebrate class III POU genes are widely expressed in the embryonic and adult central nervous system, where they act as transcriptional regulators of cell- and/or region-specific gene expression. We isolated four zebrafish class III POU genes, named zp-12, zp-23, zp-47 and zp-50. In this study, we examined the developmental expression patterns of the Brn-1- and Brn-2-related zp-12, zp-23 and zp-47 genes by means of whole-mount in situ hybridization. Similarly to their mammalian orthologues, the major expression site of all zebrafish zp genes is the CNS. Neurectodermal expression was first detected at the beginning of somitogenesis in spatially restricted segment-like domains in different parts of the neural plate. During somitogenesis transcript distributions changed from highly restricted to widespread but nevertheless distinct patterns found in all major subdivisions of the CNS. While zp-47 expression was detected exclusively in the CNS, localized expression of zp-12 and zp-23 was also found in the pronephric primordium and in cell clusters within the mandibular and hyoid arches. Furthermore, zp-23 transcripts were transiently detected in a restricted region of the paraxial mesendoderm and, at late embryogenesis stages, in the auditory vesicles. The early regionalized expression of all three zp genes is compatible with roles in regional specification of the neural plate. Comparison of the distinct yet overlapping expression of zp-12, zp-23, zp-47 and the previously characterized zp-50 gene implies both unique, as well as redundant functions for each family member. We propose that coordinate expression of particular combinations of class III POU genes contribute to pattern formation or cell fate determination in the developing CNS and other structures. PMID- 10842362 TI - Formation of a full complement of cranial proprioceptors requires multiple neurotrophins. AB - Inactivation of neurotrophin-3 (NT3) completely blocks the development of limb proprioceptive neurons and their end organs, the muscle spindles. We examined whether cranial proprioceptive neurons of the trigeminal mesencephalic nucleus (TMN) require NT3, brain-derived neurotrophic factor (BDNF) or neurotrophin-4 (NT4) for their development. Complements of TMN neurons and masticatory muscle spindles were decreased by 62% in NT3 null mutants, 33% in BDNF null mutants, and 10% in NT4 null mutant mice at birth. The extent of proprioceptive deficiencies differed among different masticatory muscles, particularly in NT3 null mice. Masticatory muscles of embryonic mice heterozygous for the NT3(lacZneo) or BDNF(lacZ) reporter genes expressed both NT3 and BDNF, consistent with target derived neurotrophin support of TMN neurons. Although more than 90% of TMN neurons expressed TrkB as well as TrkC receptor proteins by immunocytochemistry in wild-type newborns, TrkC or TrkB null mice exhibited only partial proprioceptive deficiencies similar to those present in NT3 or BDNF;NT4 null mice. Thus, in terms of the survival outcome, two main subpopulations of TMN neurons may exist during embryogenesis, one dependent on TrkC/NT3 functioning and the other utilizing TrkB/BDNF signaling. The differential dependence of TMN neurons on neurotrophins may reflect differential accessibility of the neurons to limiting amounts of NT3, BDNF, or NT4 in target tissues, especially if the tissue distribution or levels of BDNF, NT3, and NT4 were dynamically regulated both spatially and temporally. PMID- 10842363 TI - Angioblast differentiation is influenced by the local environment: FGF-2 induces angioblasts and patterns vessel formation in the quail embryo. AB - The embryonic vasculature forms by the segregation, migration, and assembly of angioblasts from mesoderm, a process termed vasculogenesis. The initial role of fibroblast growth factor 2 (FGF-2) in vascular development appears to be in the induction of endothelial precursors, angioblasts. Quail somites transplanted into chick embryos will give rise to angioblasts of quail origin. The number of angioblasts present within the chimera is dependent on the host environment. Angioblast induction can be demonstrated in vitro by the addition of FGF-2 to cultures of dissociated somitic mesoderm, as assessed by QH-1 epitope expression. Manipulation of FGF-2 concentration in the quail/chick chimeras by FGF-2 peptide or neutralizing antibody injections increases or decreases angioblast induction in the predicted manner. To better control growth factor release in vivo we have implanted beads that release FGF-2 into the embryonic environment. FGF-2 beads implanted into the somite induce angioblast differentiation in the epithelial somite; whereas, beads lateral to the somitic mesoderm induce the formation of ectopic vessels. These studies suggest that FGF-2 is important for both the induction of angioblasts and the assembly of angioblasts into the initial vasculature pattern. PMID- 10842364 TI - Requirement for BMP and FGF signaling during cardiogenic induction in non precardiac mesoderm is specific, transient, and cooperative. AB - We previously reported that combined treatment with bone morphogenetic protein-2 (BMP-2) and fibroblast growth factor-4 (FGF-4) induces cardiogenic events culminating in full cardiac differentiation of non-precardiac mesoderm explanted from stage 6 avian embryos (Lough et al. [1996] Dev. Biol. 178:198-202.). To elucidate the respective functions of BMP and FGF in initiating and maintaining the cardiogenic process, we have used these ectopic cells as a cardiac specification model to ascertain requirements for growth factor specificity and extent of application, as well as induction of cardiac transcription factors. The inability of some BMP isoforms to replace the inductive activity of BMPs-2/4 indicated a specific requirement for this signaling pathway; moreover, neither activin-A nor insulin, which support terminal differentiation of precardiac mesoderm, nor leukocyte inhibitory factor (LIF), which promotes hypertrophy in cardiac myocytes, could replace BMP's cardiogenic activity. A similarly specific requirement for FGF-2/4 signaling was revealed since neither FGF-7, activin-A nor insulin could replace this activity. The effect of both factors was concentration dependent; maximal incidence of explant differentiation for each occurred at 50 ng/ml. Surprisingly, the majority of explants treated with high BMP levels (250 ng/ml) exhibited a non-cardiac phenotype that was characterized by intense expression of alkaline phosphatase, suggesting differentiation toward an alternative mesodermal phenotype. Experiments to assess the duration of exposure to each factor that was required revealed that while exposure to BMP and FGF during only the initial 30 min of a 48-hr culture period was sufficient to induce cardiogenesis in a significant percentage of explants, 100% incidence of explant differentiation was obtained only when FGF treatment was restricted to the first 30 min and BMP was continuously present during the 48-hr culture period. Treatment with both growth factors was required to induce the cardiac transcription factors cNkx-2.5 and SRF; neither mRNA was induced by BMP or FGF alone. These findings indicate that: (1) specific members of the BMP and FGF families are required to induce cardiogenesis in non-precardiac mesoderm; (2) BMPs-2/4 may function as a morphogen; (3) brief application of both factors can induce cardiogenesis in a modest number of explants whereas (4) 100% incidence of explant differentiation can only be attained by brief FGF treatment combined with continuous BMP treatment and (5) both factors are necessary to induce downstream cardiac transcription factors. These findings are interpreted in terms of these factors' possible roles during cardiac specification and differentiation. PMID- 10842365 TI - Col2-GFP reporter marks chondrocyte lineage and chondrogenesis during mouse skeletal development. AB - Mice were generated in which a Col2-GFP transgene serves as a reporter for the chondrocyte lineage and for chondrogenesis in live embryos and newborn pups. Cells actively engaged in chondrogenesis were identified by confocal optical sectioning within their native environments in embryos and in thick tissue slices. Chondrocytes exhibiting GFP fluorescence were purified from rib cages by high-speed cell sorting of crude cell suspensions. Intensity of fluorescence correlated with biosynthesis of procollagen II in these cells. The use of these mice and their cells provides a novel approach for studying chondrocyte differentiation and chondrogenesis during skeletal development. PMID- 10842366 TI - Introduction PMID- 10842367 TI - Integrating centrosome structure with protein composition and function in animal cells. AB - The centrosome found in animal cells is a complex and dynamic organelle that functions as the major microtubule organizing center. Structural studies over the past several decades have defined the primary structural features of the centrosome but recent studies are now beginning to reveal structural detail previously unknown. Concurrent with these studies has been an explosion in the identification of the proteins that reside within the centrosome. Our growing understanding of how protein composition integrates with centrosome structure and hence with function is the focus of this review. PMID- 10842368 TI - From fertilization to cancer: the role of centrosomes in the union and separation of genomic material. AB - Centrosomes play crucial roles in the union of sperm and egg nuclei during fertilization and in the equal separation of genomic material during cell division. While many studies in recent years have focused on the molecular composition of centrosomes, this article focuses on the structural behavior of centrosomes and on factors that play a role in centrosome functions under normal, artificially altered, and abnormal conditions. We review here how studies in the classic sea urchin egg model have contributed to our knowledge on the centrosome cycle within the cell cycle, on compaction and decompaction of centrosomal material, and on the contributions of maternal and paternal centrosomes during fertilization. Centrosome material is activated in unfertilized eggs by increasing pH with ammonium and by increasing calcium with the ionophore A23187, which are conditions that are normally induced by sperm. D(2)O and taxol also induce centrosome aggregation in the unfertilized egg. Maternal and paternal centrosome material both contribute to the formation of a functional centrosome but the formation of a bipolar centrosome requires material from the paternal centrosome. Fertilization of taxol-treated eggs reveals that the male centrosome possesses the capability to attract maternal centrosome material. When pronuclear fusion of the male and female pronuclei is inhibited with agents such as the disulfide reducing agent dithiothreitol (DTT) a bipolar mitotic apparatus is formed from the paternal centrosome. Furthermore, one centrosome of the bipolar mitotic apparatus is capable of organizing an additional half spindle that attaches to the female pronucleus indicating a functional and perhaps structural connection between centrosomes and chromatin. Sea urchin eggs are also useful to study centrosome abnormalities and consequences for the cell cycle. While classic studies by Theodor Boveri have shown that dispermic fertilization will result in abnormal cell division because of multiple centrosomes contributed by sperm, abnormal cell division can also be induced by chemical alterations of centrosomes. Compaction and decompaction of centrosome structure is studied using chloral hydrate or the chaotropic agent formamide, which reveals that centrosomes can be chemically altered to produce mono- or multipolar abnormal mitosis and unequal distribution of genomic material upon release from formamide. The patterns of abnormal centrosome reformations after recovery from formamide treatment resemble those seen in cancer cells which argues that structural defects of centrosomes can account for the formation of abnormal mitosis and multipolar cells frequently observed in cancer. In summary, the sea urchin model has been most useful to gain information on the role of centrosomes during fertilization and cell division as well as on adverse conditions that play a role in centrosome dysfunctions and in disease. PMID- 10842369 TI - Centrosome precursors in the acentriolar mouse oocyte. AB - Centrioles disappear from the mouse oocyte during early oogenesis. However, it has been known for some time that multiple structures known as microtubule organizing centers (MTOC) form the spindle poles during meiosis as well as the mitotic poles during early cleavage. The objective of this study was to identify and describe the structures which exist prior to the appearance of the multiple MTOC associated with meiotic division. Reported here for the first time is a description of the unique structures which exist before the onset of oocyte maturation, their location, and microtubule (MT) nucleating ability. Correlative confocal, immuno-, and electron microscopic studies of mouse oocytes released from ovaries directly into 2% paraformaldehyde (time zero) show two large gamma tubulin-positive structures in the cortex, averaging 10 microm in diameter. The present work is the first to demonstrate that although these structures contain gamma-tubulin, they do not resemble MTOC morphologically nor do they appear to nucleate MT. They are termed here multivesicular aggregates (MVA), and ultrastructural analysis reveals that they contain a variety of vesicular structures including many ring structures of approximately 25 nm. At the onset of maturation, these two MVA migrate toward the GV breaking into smaller units, only some of which mature into MTOC and nucleate MT. These correlative microscopic studies support the conclusion that MVA are centrosomal precursors, but with a unique ultrastructure. The ultrastructural organization of MVA may explain the cryptic function of MTOC in the prematuration environment of the dictyate oocyte. PMID- 10842371 TI - Centrosome inheritance in sheep zygotes: centrioles are contributed by the sperm. AB - The inheritance and duplication of the sperm centriole in the sheep zygote was studied by transmission electron microscopy. We found two centrioles at one pole and a single centriole at the opposite pole of the first mitotic spindle, in monospermic eggs, 20-21 hours postinsemination. This indicated both duplication and relocation of centrioles to opposite spindle poles during fertilization. The absence of centrioles in mature sheep oocytes was confirmed. Following activation by the calcium ionophore A 23187, mature oocytes entered mitosis and formed a bipolar spindle 18 hours later. Centrioles were not detected in the mitotic spindle of parthenogenotes. Androgenetic eggs were obtained by excision of the anaphase II/telophase II meiotic spindle of fertilized eggs. They were capable of undergoing mitosis and formed one or two bipolar spindle(s) in monospermic and dispermic eggs, respectively, 20-24 hours postinsemination. In two monospermic androgenetic eggs, two centrioles were found at one pole and a single centriole at the opposite pole of the first mitotic spindle. Three centrioles were also observed in another androgenetic egg in prometaphase of the first mitotic division, in close vicinity to the sperm neck-piece. These data provide evidence that the sperm centriole do reproduce and occupy a pivotal position on opposite spindle poles at syngamy. Altogether, the present findings suggest that centrioles of sheep zygotes are paternally derived. PMID- 10842370 TI - Sorting and reorganization of centrosomes during oocyte maturation in the mouse. AB - In animal oocytes, the centrosome exists as an acentriolar aggregate of centrosomal material that is regulated in a dynamic manner throughout the process of meiotic maturation. Recently, it has been demonstrated that in female meiotic systems spindle assembly is likely regulated by chromosomal and microtubule/microtubule-associated influences. The purpose of this study was to analyze the distribution of the integral centrosomal protein, pericentrin, during the course of meiotic maturation. The function of the centrosome during meiotic progression was evaluated by exposing oocytes to pharmacological agents that perturb cytoplasmic homeostasis (cycloheximide, nocodazole, cytochalasin D, taxol, and vanadate). Pericentrin was localized to the spindle poles during metaphase of meiosis-I as O- and C-shaped structures. At anaphase, these structures fragment, become displaced from the spindle poles, and associate with the lateral spindle margin. The metaphase spindle at meiosis-II had incomplete pericentrin rings at both spindle poles. Vanadate treatment, a known inhibitor of dynein-ATPase, resulted in meiotic arrest, constriction of the spindle pole, and an aggregation of pericentrin at the spindle poles. After taxol exposure, pericentrin incorporation into both spindle poles and cytoplasmic centrosomes was increased. Treatment of oocytes with cycloheximide, nocodazole, and cytochalasin D, influenced early events associated with chromosome capture and spindle assembly and altered the number and distribution of cytoplasmic centrosomes. Thus, although pericentrin incorporation is not required for meiotic spindle formation, the dynamic reorganization of pericentrin and changes in centrosome microtubule nucleating capacity are involved in critical cell cycle transitions during meiotic maturation. PMID- 10842372 TI - GFP-centrin as a marker for centriole dynamics in living cells. AB - A long-standing puzzle in cell biology is the question of how cells generate one and only one new centrosome in each cell cycle and what is the role of the centriole pair in this process. In this study, the introduction of GFP-centrin into cultured cells allows direct visualization of centriole behavior in living cells and in real time. Using this method, centriole dynamics can be observed throughout the cell cycle and following a variety of experimental treatments. Our studies demonstrate that the biogenesis of new centrioles from individual members of a preexisting centriole pair is asynchronous: the older centriole initiates assembly of a new daughter centriole before the younger centriole initiates assembly of its daughter. PMID- 10842373 TI - Cell cycle-dependent phosphorylation of centrosomes: localization of phosphopeptide specific antibodies to the centrosome. AB - The microtubule nucleation capacity of the centrosome increases dramatically as cells progress from interphase into mitosis. The increase in nucleation capacity of the centrosome correlates with the cell cycle-dependent localization of the mitotic protein monoclonal-2 (MPM-2) phosphoepitope-specific antibody to the mitotic centrosome. Therefore, the phosphorylation state of centrosomal components may regulate the microtubule nucleation capacity of this organelle during mitosis. Neither the identity of the MPM-2 kinase(s) nor all of the MPM-2 reactive phosphoproteins associated with the centrosome have been fully elucidated. Only recently have the characteristics of the MPM-2 epitope site been defined, and we used this information to prepare polyclonal antibodies against synthetic phosphopeptides containing potential MPM-2 epitopes derived from the sequences of two MPM-2-reactive proteins, topoisomerase II, and microtubule associated protein 1B (MAP1B). We demonstrate that these phosphopeptide-specific antibodies also localize to the centrosome in a cell cycle-dependent fashion. Thus, polyclonal antibodies have been generated against defined phosphopeptides that reiterate many of the immunofluorescence staining properties exhibited by the MPM-2 antibody. These new phosphopeptide-specific antibodies will provide additional probes to examine the phosphorylation of centrosomal components and the functional consequences of their phosphorylation during mitosis. PMID- 10842374 TI - NuMA: a nuclear protein involved in mitotic centrosome function. AB - Nuclear mitotic apparatus protein, NuMA, is an abundant 240 kDa protein with microtubule (MT) binding capacity via its carboxyl terminal region. Structurally, it has been shown to be a double-strand coiled-coil that has a high potential to form filamentous polymers. During interphase, NuMA locates within the nucleus but rapidly redistributes to the separating centrosomes during early mitosis. Xenopus NuMA associates with MT minus end-directed motor cytoplasmic dynein and its motility-activating complex dynactin at mitotic centrosomal regions. This NuMA motor complex binds the free ends of MTs, converging and tethering spindle MT ends to the poles. A similar scenario appears to be true in higher vertebrates as well. As a mitotic centrosomal component, NuMA is essential for the organization and stabilization of spindle poles from early mitosis until at least the onset of anaphase. The cell cycle-dependent distribution and function of NuMA is regulated by phosphorylation and dephosphorylation, and p34/CDC2 activity is important to the mitotic role of NuMA. This review summarizes data about the structural features and mitotic function of NuMA with particular emphasis on the newly discovered NuMA-motor complex in spindle organization. Furthermore, NuMA may represent a large group of proteins whose mitotic function is sequestered in the nucleus during interphase. PMID- 10842375 TI - Filamentous polymers induced by overexpression of a novel centrosomal protein, Cep135. AB - A novel 135 kDa centrosomal component (Cep135) was identified by immunoscreening of a mammalian expression library with monoclonal antibodies raised against clam centrosomes. It is predicted to be a highly coiled-coil protein with an extensive alpha-helix, suggesting that Cep135 is a structural component of the centrosome. To evaluate how the protein is arranged in the centrosomal structure, we overexpressed Cep135 polypeptides in CHO cells by transient transfection. HA- or GFP-tagged full (amino acids 1-1144) as well as truncated (#10, 29-1144; Delta3, 29-812) polypeptides become localized at the centrosome and induce cytoplasmic dots of various size and number in CHO cells. Centrosomes are associated with massive approximately 7 nm filaments and dense particles organized in a whorl like arrangement in which parallel-oriented dense lines appear with a regular approximately 7 nm periodicity. The same filamentous aggregates are also detected in cytoplasmic dots, indicating that overexpressed Cep135 can assemble into elaborate higher-ordered structures in and outside the centrosome. Sf9 cells infected with baculovirus containing Cep135 sequences induce filamentous polymers which are distinctive from the whorl seen in CHO cells; #10 forms highly packed spheroids, but the Delta3-containing structure looks loose. Both structures show an internal repeating unit of dense and less dense stripes. Although the distance between the outer end of two adjacent dense lines is similar between two types of polymers ( approximately 120 nm), the dense stripe of Delta3 polymers ( approximately 40 nm) is wider than #10 ( approximately 30 nm). The light band of Delta3 ( approximately 40 nm) is thus narrower than #10 ( approximately 60 nm). Since thin fibers are frequently seen to extend from one dense line to the next, the coiled-coil rod of Cep135 may span the light band. These results suggest that overexpressed Cep135 assemble into distinctive polymers in a domain-specific manner. PMID- 10842376 TI - Higher plant cells: gamma-tubulin and microtubule nucleation in the absence of centrosomes. AB - The assembly of the higher plant cytoskeleton poses several fundamental questions. Since different microtubule arrays are successively assembled during the cell cycle in the absence of centrosomes, we can ask how these arrays are assembled and spatially organized. Two hypotheses are under debate. Either multiple nucleation sites are responsible for the assembly and organization of microtubule arrays or microtubule nucleation takes place at one site, the nuclear surface. In the latter case, microtubule nucleation and organization would be two distinct but coregulated processes. During recent years, novel approaches have provided entirely new insights to understand the assembly and dynamics of the plant cytoskeleton. In the present review, we summarize advances made in microscopy and in molecular biology which lead to novel hypotheses and open up new fields of investigation. From the results obtained, it is clear that the higher plant cell is a powerful model system to investigate cytoskeletal organization in acentrosomal eukaryotic cells. PMID- 10842378 TI - Arizona imaging and microanalysis society PMID- 10842377 TI - Microtubule organizing centers and the origin of centrioles during spermatogenesis in the pteridophyte Phylloglossum. AB - Spermatogenesis in the lycophyte Phylloglossum is characterized by profound ultrastructural changes that involve complex microtubule arrays and discrete microtubule organizing centers (MTOCs). The first visible MTOC is an electron opaque acentriolar centrosome that organizes the mitotic spindles in late spermatogeneous cells. In the spermatid mother cell, centrioles arise de novo within the pericentriolar matrix of the MTOC. Approximately 20 centrioles, which ultimately function as basal bodies, originate in each of two branched "blepharoplasts." Constituent centrioles of each organelle radiate from a central region where they are interconnected by cartwheel cylinders, each with nine-fold symmetry. The development and structure of this novel centriolar-generating organelle suggests a direct evolutionary link with the bicentriole of other lycophytes, and are consistent with the concept that multiflagellated spermatozoids in Phylloglossum evolved independently of those in other pteridophytes. During spermiogenesis, two additional structurally defined MTOCs organize the ton and locomotory apparatus, which comprises 20 staggered flagella over a multilayered structure. An MTOC that overlies the multilayered structure and consists of a cloud of electron-opaque material is involved in repositioning basal bodies and generating flagella. The spline, a band of up to 200 microtubules, provides the architectural framework for development and maintenance of cell shape and is organized by the lamellar strip, a highly structured MTOC. Microtubule arrays during spermatogenesis in Phylloglossum are diverse and include mitotic, cytokinetic, cytoskeletal, and locomotory assemblages. MTOCs responsible for the nucleation and organization of these arrays are among the most elaborate and morphologically distinct of any described in land plants. PMID- 10842379 TI - First FDA approval under humanitarian device exemption of a septal occluder for fenestrated fontan and muscular ventricular septal defects. PMID- 10842380 TI - Global experience in cervical carotid artery stent placement. AB - The purpose of this article is to review and update the current status of carotid artery stent placement in the world. Surveys to major interventional centers in Europe, North and South America, and Asia were initially completed in June 1997. Subsequent information from these 24 centers in addition to 12 new centers has been obtained to update the information. The survey asked the various questions regarding the patients enrolled, procedure techniques, and results of carotid stenting, including complications and restenosis. The total number of endovascular carotid stent procedures that have been performed worldwide to date included 5,210 procedures involving 4,757 patients. There was a technical success of 98.4% with 5,129 carotid arteries treated. Complications that occurred during the carotid stent placement or within a 30-day period following placement were recorded. Overall, there were 134 transient ischemic attacks (TIAs) for a rate of 2.82%. Based on the total patient population, there were 129 minor strokes with a rate of occurrence of 2.72%. The total number of major strokes was 71 for a rate of 1.49%. There were 41 deaths within a 30-day postprocedure period resulting in a mortality rate of 0.86%. The combined minor and major strokes and procedure related death rate was 5.07%. Restenosis rates of carotid stenting have been 1.99% and 3.46% at 6 and 12 months, respectively. The rate of neurologic events after stent placement has been 1.42% at 6-12-month follow-up. Endovascular stent treatment of carotid artery atherosclerotic disease is growing as an alternative for vascular surgery, especially for patients that are high risk for standard carotid endarterectomy. The periprocedure risks for major and minor strokes and death are generally acceptable at this early stage of development and have not changed significantly since the first survey results. Cathet. Cardiovasc. Intervent. 50:160-167, 2000. PMID- 10842381 TI - A world of promise--but more to do. PMID- 10842382 TI - Nine-year follow-up of balloon-expandable Palmaz-Schatz stent in patients with single-vessel disease. AB - The long-term effects of intracoronary stents in human are unknown. This is the first 9-year follow-up report of single-vessel-disease patients treated with the Palmaz-Schatz stent. Between March and December 1989, out of the 107 patients undergoing Palmaz-Schatz stent implantation, 71 (66%) had single-vessel disease. The average age of these patients was 58+/-9 years and 79% were men. At 9 years, follow-up was obtained for 90.1% and major adverse clinical events consisted of 4 deaths giving a global survival rate of 95.8%, 7 myocardial infarction, 3 bypass surgeries, and 16 repeat percutaneous revascularization procedures. The 9-year event-free survival rate was 60%, and 81.7% of the patients were free from death, myocardial infarction, and bypass surgery. Multivariate analysis showed that the only predictive factor of major adverse clinical events was the presence of diabetes mellitus (P<0.004). Cathet. Cardiovasc. Intervent. 50:170-174, 2000. PMID- 10842383 TI - On the resilience of coronary stents: are they timeless or do they corrode? PMID- 10842384 TI - QRS prolongation: a sensitive marker of ischemia during percutaneous transluminal coronary angioplasty. AB - The purpose of this study is to measure QRS duration changes in the human model of ischemia during percutaneous transluminal coronary angioplasty (PTCA) and compare these results to the commonly used ischemia markers, chest pain, and classical ST-T changes. Using a computerized method, QRS duration was measured in 51 patients undergoing elective PTCA. Three milliseconds (msec) or more prolongation of the QRS at peak inflation was considered to be an ischemic response. The results were compared to chest pain and ST-T changes and were analyzed for inflation site within individual coronary arteries. Forty-two patients had a pathological prolongation of the QRS during PTCA. Thirty-two patients developed chest pain, while 19 had ischemic ST-T changes. QRS duration was more prolonged in PTCA to proximal or middle segments of major arteries or their large branches, while it was less prolonged in distal segments or smaller branches. Using our method, QRS prolongation was an ischemia marker in most patients during PTCA and was more sensitive than chest pain or ST-T changes. QRS duration was more prolonged with occlusion of proximal and middle segments of major arteries. Cathet. Cardiovasc. Intervent. 50:177-183, 2000. PMID- 10842385 TI - Stenting: the long and the short of it, or see spot run? PMID- 10842386 TI - Giant aneurysm following coil occlusion of patent ductus arteriosus. AB - A case is described in which a giant aneurysm developed following successful PDA coil occlusion in an infant with Marfan syndrome. This rare and severe complication brings into question the need for careful evaluation of these children before and after transcatheter occlusion. PMID- 10842387 TI - Aneurysm of the patent ductus arteriosus after coil closure: a rare complication. PMID- 10842388 TI - Anterograde coil closure of patent ductus arteriosus using a modified bioptome delivery technique. AB - Three consecutive patients are presented who underwent successful anterograde catheter coil occlusion of a patent ductus arteriosus using a modified bioptome assisted technique. Two of the three patients were infants and the procedures were performed without the need for arterial access. PMID- 10842390 TI - Interventional PFO closure: what we see is but the tip of the iceberg. PMID- 10842389 TI - Closure of patent foramen ovale in patients with orthodeoxia-platypnea using the amplatzer devices. AB - We present a series of four patients with orthodeoxia-platypnea who underwent successful transcatheter closure of their patent foramen ovale using the Amplatzer devices (Amplatzer septal occluder/Amplatzer PFO occluder). The average saturation increased from 81% to 96% with complete resolution of symptoms. The Amplatzer devices are safe and effective treatment options for patients with orthodeoxia-platypnea. PMID- 10842391 TI - Five-loop coil occlusion of patent ductus arteriosus prevents recurrence of shunt at follow-up. AB - Recent reports suggest reopening of the patent ductus arteriosus (PDA) after complete occlusion with three-loop Gianturco coils. We hypothesize that five-loop coils may produce a larger thrombus than three-loop coils, which will result in no or less probability of recanalization of PDA during follow-up. This study is designed to test this hypothesis. Follow-up echocardiographic and Doppler data of 30 patients who underwent five-loop coil occlusion of small to medium-sized PDA during a 33-month period ending December 1998 were examined. Thirty patients had no residual shunt on echo Doppler study on the day following the procedure and were followed for 6 to 30 months (median, 12) after coil implantation. At the last follow-up study, none of the patients had a residual shunt and left atrial size decreased. Careful pulsed, continuous wave, and color Doppler interrogation of left/main pulmonary artery junction and proximal descending aorta did not reveal any evidence for obstruction. The follow-up data suggest that complete occlusion of small- to medium-sized PDAs is feasible with five-loop coils without evidence for recanalization at a mean follow-up of 12 months. Much longer (2 to 5 years) follow-up data may be necessary to confirm these observations. We speculate that a greater degree of thrombosis is produced within the ductus by the five-loop coils, which in turn may be responsible for lack of shunt recurrence. We recommend use of five-loop instead of three-loop coils for transcatheter occlusion of small- to medium-sized PDAs. PMID- 10842392 TI - Treatment of pulmonary artery stenosis after arterial switch operation: stent implantation vs. balloon angioplasty. AB - The development of pulmonary artery stenosis is a potential complication during the mid- to long-term follow-up after arterial switch operation (ASO) for transposition of the great arteries. Surgical results have been disappointing and conventional balloon dilation yields a fairly important incidence of failures and recurrences. We evaluated our results with implantation of balloon-expandable stents in 5 out of 13 patients with a previously attempted unsuccessful conventional balloon dilation of pulmonary artery branch stenosis after ASO. In two more cases, stents were used as a primary procedure. Balloon angioplasty achieved a 15% increase in mean diameter of the stenosis vs. 124% with the use of stents (P< 0.01), a 10% decrease of the pressure gradient across the stenosis vs. 71% of stents (P<0.01), and a 10% drop in RV/aorta pressure ratio vs. 43% of stents (P<0.01). Compared to conventional balloon angioplasty in our series, stents were more effective in the treatment of patients with peripheral pulmonary artery stenosis after ASO. Balloon dilation should be considered in selected cases unsuitable for treatment with endovascular stents. PMID- 10842393 TI - Repeated balloon rupture during coronary stenting due to a calcified lesion: an intravascular ultrasound study. AB - We describe a patient in whom balloon rupture occurred three times during inflation in a stent with restenosis in the left anterior descending artery. The cause of rupture was detected by intravascular ultrasound: a calcified ridge that protruded between the stent struts in the distal stent body. PMID- 10842394 TI - Facilitation of stent retention and retrieval with an emboli containment device. AB - A potential consequence of unsuccessful stent delivery is dislodgment of the stent from the delivery catheter resulting in embolization. Recently, an angioplasty guidewire incorporating a distal occlusion balloon (GuardWire) has become available. We describe how, when used for the prevention of distal embolization of atheromatous or thrombotic particles, this device may facilitate retention and retrieval of undeployed stents. PMID- 10842395 TI - Patience in the pursuit of perfection. PMID- 10842396 TI - Pressure wire kinking, entanglement, and entrapment during intravascular ultrasound studies: a potentially dangerous complication. AB - The simultaneous use of intravascular ultrasound catheters and sensor-tipped guidewires is gaining acceptance during coronary interventions as a means to gain further insights on the significance of coronary stenoses. Herein we describe four patients in whom the distal tip of the pressure wire became entrapped during an intravascular ultrasound examination. In the four patients, a localized kinking of the pressure wire initially prevented the removal of the imaging catheter and eventually the wire-catheter assembly had to be retrieved as a unit into the guiding catheter. In one patient, unraveling of the distal part of the pressure wire was noticed. In two patients, a complete loop with further kinking of the pressure wire was induced during the maneuvers performed to withdraw the imaging system. Three patients experienced transient angina. Although in our patients this technical problem was not associated with any clinical sequelae, interventional cardiologists should be aware of the potential complications associated with the combined use of these two intracoronary diagnostic tools. PMID- 10842397 TI - Primary cardiac lymphoma diagnosed by percutaneous intracardiac biopsy with combined fluoroscopic and transesophageal echocardiographic imaging. AB - Primary cardiac tumors have very low prevalence with cardiac lymphoma, being one of the rarest forms. Several recent reports have shown transesophageal echocardiography to be an accurate technique for characterizing and localizing these neoplasms, with results comparable to CT and MRI scans. Transvenous intracardiac tumor biopsy has been employed as a minimally invasive technique to obtain tissue samples. The addition of transesophageal echocardiographic (TEE) guidance to this process has increased the accuracy of obtaining diagnostic specimens while improving patient safety. We review published cases of this relatively new technique using combined fluoroscopic and TEE guidance and present a case of primary cardiac lymphoma diagnosed by this method. The patient achieved complete tumor remission after treatment with standard chemotherapy and remains fully functional 32 months after initial diagnosis. PMID- 10842398 TI - Congenital arteriovenous fistulas of the bilateral internal mammary vessels. AB - Internal mammary artery-to-venous fistulas represent a rare cause of a continuous precordial murmur that may be confused clinically with a patent ductus arteriosus. The acquired variety may be secondary to inflammatory, neoplasic, or traumatic events. A congenital fistula may occur as an isolated event, as in the present case, and the question of advisability of endovascular treatment for this patient remains unanswered. PMID- 10842399 TI - Hemodynamic rounds series III: hemodynamics of the human left atrial appendage. PMID- 10842400 TI - Current perspectives on interventional treatment strategies in diabetic patients with coronary artery disease. AB - Diabetes mellitus has a negative impact on mortality and morbidity following catheter-based coronary procedures as well as coronary artery bypass surgery. Increased restenosis remains the main limitation of catheter-based coronary intervention among diabetes mellitus in addition to accelerated atherosclerosis lesion progression in other untreated coronary sites. Determinants such as excess restenosis, high atherosclerosis burden, lesion complexity, small target vessel size, and accelerated coronary atherosclerosis in remote sites may favor the surgical strategy in most cases of diabetic multivessel disease. The importance of periprocedural adjunctive pharmacotherapy, specifically with the use of antiplatelet and long-term antilipidemic treatment, was shown to improve outcomes in diabetics undergoing percutaneous coronary interventions. The purpose of the review is to examine potential mechanisms causing more restenosis in diabetics, the clinical outcomes of patients with diabetes after coronary interventions including stenting, the treatment alternatives of diabetic patients with diffuse coronary artery disease, including coronary bypass surgery, and current understanding of the benefit of adjunctive pharmacology on clinical outcomes after coronary interventions among diabetics. PMID- 10842401 TI - Conversion of supra-ventricular tachycardia by catheter manipulation of the right atrium. AB - A case is described by which the right heart catheter was utilized to convert mechanically an episode of supraventricular tachycardia. PMID- 10842402 TI - Image quality assessment in contemporary interventional cardiology laboratories: spatial and low-contrast video resolution. AB - Two measures of fluoroscopic image quality-high-contrast video spatial resolution and low-contrast video resolution-were assessed in 64 interventional cardiology catheterization laboratories. The results of high-contrast spatial resolution testing indicate variable levels of performance with median values of 1.5, 2.0, and 2.5 line pairs/mm at the three levels of magnification. The results of the low-contrast video resolution testing indicate clinically relevant limitations in performance. This was particularly true when testing was performed in a dynamic mode under simulated large patient conditions. Under these conditions, 50% of systems failed to detect a wire of diameter 0.017". In addition, limited system dynamic range was identified in a substantial fraction of laboratories and may contribute to these important variations in performance. Taken together, these findings serve to characterize overall system performance in the current era of interventional cardiology and reaffirm the continued need for image quality assessment programs. PMID- 10842403 TI - Why measure imaging system performance? PMID- 10842404 TI - Early clinical experience with the implantation of a novel synthetic coronary stent graft. PMID- 10842405 TI - A word of caution on unrestricted use of synthetic stent grafts in native coronary arteries PMID- 10842406 TI - Guidelines for ad-hoc coronary intervention: role of physiologic justification. PMID- 10842407 TI - Talk to Us PMID- 10842408 TI - Muscle force and range of motion as predictors of function in older adults. AB - BACKGROUND AND PURPOSE: Musculoskeletal impairments and functional limitations are linked to disability in older adults. The purposes of this study were to identify the extremity musculoskeletal impairments that best predict functional limitations in older adults and to assess the validity of measurements obtained for the Physical Performance Test (PPT) as a predictor of disability. SUBJECTS AND METHODS: Eighty-one older adults residing in independent and dependent care facilities were tested for extremity muscle force, range of motion, and function. Data were analyzed using multiple regression analysis to identify extremity impairments that predicted function scores and logistic regression analysis to determine whether PPT scores predicted subjects' living situation as dependent versus independent. RESULTS: Subject age, lower-extremity muscle force, and lower extremity range of motion explained 77% of the variance in function as measured by the PPT. Results differed when analysis was done by subject living situation, with a higher percentage of the variance in function scores explained by musculoskeletal measures for the dependent living group as compared with the independent living group. CONCLUSION AND DISCUSSION: Extremity musculoskeletal impairments have a strong relationship to function, especially in older adults living in dependent care settings. The results of this study can be used to design interventions to address the musculoskeletal disorders most related to function in the older population. PMID- 10842409 TI - Abdominal muscle response during curl-ups on both stable and labile surfaces. AB - BACKGROUND AND PURPOSE: With the current interest in stability training for the injured low back, the use of labile (movable) surfaces, underneath the subject, to challenge the motor control system is becoming more popular. Little is known about the modulating effects of these surfaces on muscle activity. The purpose of this study was to establish the degree of modulating influence of the type of surface (whether stable or labile) on the mechanics of the abdominal wall. In this study, the amplitude of muscle activity together with the way that the muscles coactivated due to the type of surface under the subject were of interest. SUBJECTS: Eight men (mean age=23.3 years [SD=4.3], mean height=177.6 cm [SD=3.4], mean weight=72.6 kg [SD=8.7]) volunteered to participate in the study. All subjects were in good health and reported no incidence of acute or chronic low back injury or prolonged back pain prior to this experiment. METHODS: All subjects were requested to perform 4 different curl-up exercises-1 on a stable surface and the other 3 on varying labile surfaces. Electromyographic signals were recorded from 4 different abdominal sites on the right and left sides of the body and normalized to maximal voluntary contraction (MVC) amplitudes. RESULTS: Performing curl-up exercises on labile surfaces increased abdominal muscle activity (eg, for curl-up on a stable surface, rectus abdominis muscle activity was 21% of MVC and external oblique muscle activity was 5% of MVC; for curl-up with the upper torso on a labile ball, rectus abdominis muscle activity was 35% of MVC and external oblique muscle activity was 10% of MVC). Furthermore, it appears that increases in external oblique muscle activity were larger than those of other abdominal muscles. CONCLUSION AND DISCUSSION: Performing curl-ups on labile surfaces changes both the level of muscle activity and the way that the muscles coactivate to stabilize the spine and the whole body. This finding suggests a much higher demand on the motor control system, which may be desirable for specific stages in a rehabilitation program. PMID- 10842410 TI - Effects of squat lift training and free weight muscle training on maximum lifting load and isolinetic peak torque of young adults without impairments. AB - BACKGROUND AND PURPOSE: Manual lifting is a frequent cause of back injury, and there is no evidence as to which training mode can provide the best training effect for lifting performance and muscle force. The purpose of this study was to examine the effects of a squat lift training and a free weight muscle training program on the maximum lifting load and isokinetic peak torque in subjects without known neuromuscular or musculoskeletal impairments. SUBJECTS: Thirty-six adults (20 male, 16 female) without known neuromuscular or musculoskeletal impairments participated. The subjects' mean age was 21.25 years (SD=1.16, range=20-24). METHODS: Subjects were divided into 3 groups. Subjects in group 1 (n=12) performed squat lift training. Subjects in group 2 (n=12) participated in free weight resistance training of their shoulder abductors, elbow flexors, knee extensors and trunk extensors. Subjects in group 3 (n=12) served as controls. The maximum lifting load and isokinetic peak torques of the trunk extensors, knee extensors, elbow flexors, and shoulder abductors of each subject were measured before and after the study. Training was conducted on alternate days for 4 weeks, with an initial load of 80% of each subject's maximum capacity and with the load increased by 5% weekly. RESULTS: All groups were comparable for all measured variables before the study. After 4 weeks, subjects in groups 1 and 2 demonstrated more improvement in maximum lifting load and isokinetic peak torque of the back extensors compared with the subjects in group 3, but the 2 training groups were not different. CONCLUSION AND DISCUSSION: The findings demonstrate that both squat lift and free weight resistance training are equally effective in improving the lifting load and isokinetic back extension performance of individuals without impairments. PMID- 10842411 TI - Movement disorders in people with Parkinson disease: a model for physical therapy. AB - People who are diagnosed with idiopathic Parkinson disease (PD) experience movement disorders that, if not managed, can lead to considerable disability. The premise of this perspective is that physical therapy for people with PD relies on clinicians having: (1) up-to-date knowledge of the pathogenesis of movement disorders, (2) the ability to recognize common movement disorders in people with PD, (3) the ability to implement a basic management plan according to a person's stage of disability, and (4) problem-solving skills that enable treatment plans to be tailored to individual needs. This article will present a model of physical therapy management for people with idiopathic PD based on contemporary knowledge of the pathogenesis of movement disorders in basal ganglia disease as well as a review of the evidence for physical therapy interventions. The model advocates a task-specific approach to training, with emphasis on treating people with PD related movement disorders such as hypokinesia and postural instability within the context of functional tasks of everyday living such as walking, turning over in bed, and manipulating objects. The effects of medication, cognitive impairment, the environment, and coexisting medical conditions are also taken into consideration. An argument is put forward that clinicians need to identify core elements of physical therapy training that apply to all people with PD as well as elements specific to the needs of each individual. A case history is used to illustrate how physical therapy treatment is regularly reviewed and adjusted according to the changing constellation of movement disorders that present as the disease progresses. PMID- 10842412 TI - A multivariate model of determinants of motor change for children with cerebral palsy. AB - The purpose of this article is to describe the development of a theory- and data based model of determinants of motor change for children with cerebral palsy. The dimensions of human functioning proposed by the World Health Organization, general systems theory, theories of human ecology, and a philosophical approach incorporating family-centered care provide the conceptual framework for the model. The model focuses on relationships among child characteristics (eg, primary and secondary impairments, personality), family ecology (eg, dynamics of family function), and health care services (eg, availability, access, intervention options). Clarification of the complex multivariate and interactive relationships among the multiple child and family determinants, using statistical methods such as structural equation modeling, is necessary before determining how physical therapy intervention can optimize motor outcomes of children with cerebral palsy. We propose that the development and testing of multivariate models is also useful in physical therapy research and in the management of complex chronic conditions other than cerebral palsy. Testing of similar models could provide physical therapists with support for: (1) prognostic discussions with clients and their families, (2) establishment of realistic and attainable goals, and (3) interventions to enhance outcomes for individual clients with a variety of prognostic attributes. PMID- 10842413 TI - Vocational rehabilitation perspective. PMID- 10842414 TI - Too many variables, not enough equations. PMID- 10842415 TI - Different method, different results? PMID- 10842416 TI - Diuretic use: a risk for subclinical thiamine deficiency in elderly patients. AB - Long term diuretic therapy represents one central pharmacologic therapy of heart insufficiency and hypertension. Diuretics lead not only to an increased urinary excretion of electrolytes but also of water soluble vitamins. In this prospective study we evaluated the effect of hospitalization on the overall biochemical vitamin status in subjects older than 50 years (n=149, mean +/- SD age 70 +/- 10 years). Vitamin nutriture and other parameters were assessed at admission and discharge (duration of the hospitalization 19 +/- 1 day). Only vitamin B1 nutriture worsened during the hospitalization and in a multivariate procedure the only significant predictor of the change in the vitamin B1 nutriture was the use of diuretics during the hospitalization (F=4.06, p < 0.001). The changes in the ETK (erythrocyte transketolase activity in whole blood) and a-ETK (ETK activity coefficient) during the hospital stay correlated with the cumulative dosage of furosemide adjusted for the duration of the therapy (r = 0.36, p < 0.001 and r = 0.28, p > 0.03). Our data suggest that hospitalized elderly are at increased risk for vitamin B1 deficiency especially when on a diuretic treatment. It is possible that a low dose thiamine supplementation my help to prevent the development of a subclinical wet-beriberi in older subjects on diuretics. PMID- 10842417 TI - Prediction of stature in the Italian elderly. AB - Adequate quantification of weight and stature is essential in order to determine levels of nutritional support and to monitor the effects of nutritional intervention. Traditional anthropometric techniques are difficult to apply in elderly or handicapped patients chair or bed-bound. The purpose of the present study is to elaborate regression equations for the estimation of stature in the italian elderly population from other anthropometric measures that can be more easily determined. We have found a single model valid for both sexes (in which the value of the variable "sex" equals 0 if woman and 1 if man) to predict stature in italian elderly: Stature = 94.87 + 1.58 knee-height - 0.23 age + 4.8 sex. Cross validation on a control sample of 30 males and 54 females yielded pure errors of 3.1 cm for men and 2.74 cm for women. PMID- 10842418 TI - The nutritional intake of a free-living healthy French population : a four-year follow-up. AB - OBJECTIVE: To contribute to a better definition of the nutritional requirements of the healthy elderly and to improved knowledge the effects of age on these requirements. METHODS: We studied the nutritional intake of 96 elderly persons who had met criteria of good health status in 1993 at a four-year interval. The nutritional intake of the elderly subjects who remained healthy during the four year interval (18 men and 64 women), was considered to globally correspond to their nutritional needs. The nutritional intake was evaluated by a three-day food record. RESULTS: The mean baseline weight of the subjects who remained in good health during the four years was 72,6 +/- 9,5 kg for men and 60,1 +/- 9,3 kg for women. In four years, mean weight remained globally stable. But in cross sectional analysis, weight tended to decrease with the age of the subjects. This decrease was significant for women in 1993. Mean baseline intake was nearly 29 kcal/kg. Longitudinal and cross-sectional analyses showed that it changed only slightly with age. Though global energy intake varied slighlty in four years, we have observed some changes in the composition of this caloric intake. For men and women, protein intake tended to decrease in four years (respectively -0,4% and 0,6%), carbohydrate intake to decrease (respectively -1,1% and -2,7%) and fat intake to increase (respectively +2,1% and +2,9%). These variations were not significant except for fat and carbohydrate intakes in women. In 1993, for a majority of subjects, the intakes of iron, and vitamins C and B12 were higher and the intakes of calcium, zinc, vitamins A, B1, B6 and B9 were lower than the French recommendations. In four years, mean intake did not change significantly, except for calcium intake in women (-8,8%). CONCLUSION: In this study, healthy aging was associated with a mean caloric intake close to 29 kcal/kg which is near the upper limits of recommendations (between 25 and 30 kcal/kg/d). These intakes, like those of macronutrients and micronutrients changed little with advancing age. These results suggest that the needs of the elderly remain quite stable with aging. Nevertheless a tendency to weight loss with aging is observed, especially in older subjects, suggesting that even if food intake contributes to the maintenance of healthy aging, aging processes are multidimensional and frailty that is often associated with weight loss is ineluctable for older subjects. PMID- 10842419 TI - Measurement feasability of advanced glycated end-products from skin samples after antioxidant vitamin supplementation in patients with type 2 diabetes. AB - PURPOSE: To determine the feasibility of measuring advanced glycated end-products (AGEs)from skin samples and to evaluate the effects of a combination of vitamins E and C on measures of glycemic control and AGEs in patients with type 2 diabetes mellitus. METHODS: Twenty-two patients with type 2 diabetes from a Family Medicine clinic were randomly assigned to receive a daily dietary supplement containing either a combination of 400 mg of vitamin E and 500 mg of vitamin C or matching placebo for a period of one year. AGEs were measured from skin samples taken from the buttock. RESULTS: Nineteen subjects completed this one-year pilot study. There were no major problems found in measuring AGEs from skin samples taken from the butttock. Neither the treatment or placebo group had significant changes in glycemic control, protein glycosylation or AGEs. DISCUSSION: Skin samples taken from the buttock area may be an appropriate site for the determination of AGE levels as this procedure appeared to be well-tolerated. Daily vitamin E and C supplementation did not improve measures of glycemic control or AGE levels in this small sample of patients with type 2 diabetes. Because antioxidant vitamins are inexpensive and free of side effects, additional research using a variety of antioxidant vitamin combinations and dosing regimens is needed. PMID- 10842420 TI - Marginal protein intake results in reduced plasma IGF-I levels and skeletal muscle fiber atrophy in elderly women. AB - We investigated the effects of dietary protein on plasma IGF-I levels and muscle fiber cross-sectional area (CSA). Twelve healthy elderly women were randomly assigned to a weight-maintaining diet containing either 1.47 (marginal) or 2.94 (adequate) g protein/kg body cell mass (BCM)/d, (0.45 and 0.92 g/kg body weight/d, respectively) for 10wks. Plasma IGF-I levels and muscle fiber areas and distributions were evaluated at baseline and 10wks. After 10wks, both IGF-I and type I fiber CSA had declined significantly in subjects fed the marginal diet (30.1+/-2.1% and 32.7+/-7.9%, respectively) while they increased in those fed the adequate diet (19.5+/-7.0% and 22.3+/-7.5%, for IGF-I and type I CSA, respectively). The change in IGF-I was the only significantly associated with the change in type I fiber CSA (r2=0.70; p<0.03). These findings show that marginal dietary protein intakes will result in losses of muscle mass in the elderly and suggest a role for plasma IGF-I as a biochemical marker for the histological changes in skeletal muscle. PMID- 10842421 TI - Dietary intake and nutritional status in Italian elderly subjects. AB - Dietary intake and nutritional status was assessed in a random sample of 190 Italians (70-75 years of age) participating in the Survey in Europe on Nutrition and the Elderly (EURONUT-SENECA). The daily energy intake as assessed by a Modified Dietary History, was 2208+/-562 Kcalories in men and 1742+/-527 Kcalories in women. The alcohol intake was significantly higher in men than in women (35. 9+/-32.5 g/day vs 14.7+/-15.4 g/day; p<0.0001). As for calcium, there was a high percentage of men (77%) and women (86%) with a lower intake than the recommended values. The subjects underweight (BMI<20) were only 4.1% men and 9.7% women, while the great part was normal (BMI= 20-24.9) and overweight (BMI=25 29.9). The body composition parameters showed a significant difference between two genders. Men had a Total Body Water (56.5+/-4.5% vs 51.3+/-5.4%; p<0. 001) and Fat-Free Mass (80.4+/-5.2% vs 70.9+/-6.8%; p<0.001) higher than women. Few subjects were at high risk of deficiency with regard to plasma levels of vitamins, haemoglobin and albumin. If we analyse the composition of the diet consumed, we can remark the characteristics of a typical Mediterranean diet. We conclude that the general nutritional status of our sample was fairly good. PMID- 10842422 TI - Is chronic hypoxemia in patients with chronic obstructive pulmonary disease associated with more marked nutritional deficiency? A study of the fat-free mass evaluated by anthropometry and bioelectrical impedance methods. AB - In order to determine wheter blood gases abnormalities, specially hypoxemia, are associated with more marked changes in fat-free mass in patients with chronic obstructive pulmonary disease (CPOD), nutritional assessment was performed on 16 normoxemic (PaO2 > 55 mm Hg) and 16 hypoxemic (PaO2 < 55 mm Hg) COPD patients in stable clinical condition. Body weight was expressed as percentage of the ideal body weight. Fat-free mass was estimated by anthropometry (FFM-Anthr) and by bioelectrical impedance (FFM-BI). Handgrip-strength was assessed as a measure of peripheral skeletal muscle strength. Patients were age-matched and presented similar degree of airway obstruction. Malnutrition, defined as body weight less than 90% of the ideal, was observed in 19% of the normoxemic patients and in 25% of the hypoxemic patients (p>0,05). FFM values in hypoxemic patients, estimated by both methods, were not different from those observed in normoxemic patients. No significant difference was observed on handgrip values between the two groups. No correlation was found between nutritional indices and pulmonary function and gases exchange parameters. FFM correlated positively with values of peripheral muscle function in normoxemic and hypoxemic patients. These data add further evidence to the hypothesis that hypoxemia is not a primary cause of the nutritional deficiency observed in COPD patients. PMID- 10842424 TI - Preventive nutrition in old age - a review. AB - Preventive nutrition in the elderly is reviewed also from points of view of the concepts of prevention, nutrition and old age, respectively. The demographic, medical, psychosocial and other differences between individuals of the same age but belonging to different birth cohorts are emphasized, as well as the need for a multifactorial preventive approach. The interaction between dietary aspects and physical activity is discussed, and it is also pointed out that risk indicators and risk factors may be different in old age compared to middle life. Finally, public health aspects and "hospital malnutrition" are dealt with in this review. PMID- 10842423 TI - Feasibility [corrected] of regular physical exercise for patients with moderate to severe Alzheimer disease. AB - BACKGROUND: Physical activity delays loss of autonomy in the elderly. In patients with Alzheimer disease (AD), physical activity could be a useful strategy in therapeutic management by delaying loss of functional independence and the usual complications of the disease. OBJECTIVE: To determine, using standardized tools, the effects on autonomy (ADL, IADL), cognitive function (MMS), nutritional status (MNA), behavioral problems (NPI) and risk of falls (Tinetti test) of a physical exercise program in patients with AD. DESIGN: Twenty-three subjects (13 men and 10 women, aged 71-92 years, mean 78 years) with AD (mean MMS 16, range 1-23) carried out for a mean of 7 weeks (5-12 weeks) a program of endurance exercise (walking, exercise bicycle) adapted to their individual capacities. Standardized gerontological evaluation was performed before and after the study. RESULTS: No significant change in autonomy (ADL, IADL) was observed. There was an improvement in the MNA (p<0.001) and the MMS (p<0.001). Risk of falls (p<0.01) and behavioral problems (p<0.05) decreased. These results were obtained without increasing family workload. CONCLUSION: We suggest that physical activity is a therapeutic option which can reduce nutritional and behavioral complications and risk of falls in subjects with AD. PMID- 10842426 TI - Emerging adulthood. A theory of development from the late teens through the twenties. AB - Emerging adulthood is proposed as a new conception of development for the period from the late teens through the twenties, with a focus on ages 18-25. A theoretical background is presented. Then evidence is provided to support the idea that emerging adulthood is a distinct period demographically, subjectively, and in terms of identity explorations. How emerging adulthood differs from adolescence and young adulthood is explained. Finally, a cultural context for the idea of emerging adulthood is outlined, and it is specified that emerging adulthood exists only in cultures that allow young people a prolonged period of independent role exploration during the late teens and twenties. PMID- 10842425 TI - Immunonutrition - fact or fad. AB - Suggestions that modifications to the diet may have a beneficial effect on health are not new. recent work demonstrates that nutritional supplements may effect immune function. This review examines the evidence for the effects of these supplements including arginine, glutamine, nucleotides and omega - 3 fatty acids. In particular, clinical trials with patients in multiple organ failure are discussed. The evidence published thus far suggests that immunonutrition, if absorbed in adequate amounts both pre- and post-operatively, reduces septic complications and improves surrogate measures of ourcome, including length of stay and requirements for mechanical ventilation. PMID- 10842427 TI - The next wave of change for psychology and mental health services in the health care revolution. AB - Recent national changes in the de facto system of health and mental health care are described. Although the percentage of people without health insurance (always or sometimes) has not decreased, the organization of insured care has changed dramatically. Of the insured population, 75% are under some form of managed care. For 88% of the managed care population, mental health care has not been integrated with health care: the so-called carve-out. The author argues that system integration (carve-ins), for a variety of reasons, will begin soon and will occur very rapidly. A tilt toward carve-ins will have substantial impact on psychologists' training, service delivery, and research. PMID- 10842428 TI - Uncertain genesis. The academic institutionalization of American psychology in 1900. AB - The new discipline of psychology had been established at a number of American colleges and universities by 1900, but it usually existed in a more rudimentary form, as compared with the familiar autonomous department of psychology found today. The current form took quite a number of years to evolve: A century ago, a survey of these schools would have shown psychology programs to have existed mostly at early stages of development. Many of the schools were still teaching some form of moral or mental philosophy or only one or two courses in psychology. A few of the schools had established psychology laboratories. Fewer still were offering the doctor of philosophy degree in psychology, while a mere handful had independent psychology departments. PMID- 10842429 TI - Teaching the introductory course in psychology circa 1900. AB - Teachers of introductory courses in psychology and those who wrote the textbooks for the courses at the start of the 20th century represented the new psychology that replaced the old psychology of mental philosophy. Teachers and texts presented psychology as a natural science of the mind and mental processes, described its methods, and suggested its potential applications to practical concerns. Textbooks and teachers varied in their approaches to psychology, their priorities among its methods, and their emphases on applying psychology. The introductory course in psychology accurately reflected the state of the discipline at the turn of the century. PMID- 10842430 TI - A sustainable future for humanity? How can psychology help? AB - The sustainability of human life on Earth in the future is in danger. Human actions are producing many harmful and possibly irreversible changes to the environmental conditions that support life on Earth. This article summarizes major threats to Earth's environment, including global warming, ozone layer destruction, exhaustion of fisheries and agricultural land, and widespread exposure to toxic chemicals. Unless they are overcome, these changes will make human life increasingly miserable and eventually may make Earth nearly uninhabitable for future generations. These threats are caused by patterns of human behavior, particularly over-population and over-consumption. Urgent changes to human lifestyles and cultural practices are required for the world to escape ecological disaster, and psychologists should lead the way in helping people adopt sustainable patterns of living. Specific steps toward that goal are proposed in this and the following four articles. PMID- 10842431 TI - Adapting human lifestyles for the 21st century. AB - A number of ecological problems (e.g., global warming, ozone depletion, deforestation, acid rain) have been identified, which threaten to reduce the quality of human life in the 21st century. These problems are human produced, resulting primarily from over-population and over-consumption. Alterations in people's awareness, attitudes, beliefs, and behaviors may stimulate changes in their political and economic systems, which in turn might foster the kind of lifestyle changes that could mitigate these ecological problems. Psychologists can play a role in helping individuals and systems advance toward the goal of becoming a sustainable society: one that satisfies its current needs without jeopardizing the prospects of future generations. PMID- 10842432 TI - Some big ideas for some big problems. AB - Although most psychologists do not see sustainability as a psychological problem, our environmental predicament is caused largely by human behaviors, accompanied by relevant thoughts, feelings, attitudes, and values. The huge task of building sustainable cultures will require a great many psychologists from a variety of backgrounds. In an effort to stimulate the imaginations of a wide spectrum of psychologists to take on the crucial problem of sustainability, this article discusses 4 psychological approaches (neo-analytic, behavioral, social, and cognitive) and outlines some of their insights into environmentally relevant behavior. These models are useful for illuminating ways to increase environmentally responsible behaviors of clients, communities, and professional associations. PMID- 10842433 TI - Psychology and the science of human-environment interactions. AB - Psychology has an indispensable role in understanding environmental problems and finding solutions. To fill this role, psychologists must work within an interdisciplinary effort to build a scientific understanding of human-environment interactions. This article enumerates 8 widely held beliefs about these interactions and assesses the strengths and limitations of each belief. It suggests that psychology can contribute more strongly by counteracting disciplinary biases, focusing research where a behavioral analysis identifies major opportunities, making appropriately modest claims, collaborating with other disciplines, and building on psychology's relative strengths among the human sciences. PMID- 10842434 TI - Fostering sustainable behavior through community-based social marketing. AB - Psychology has a central role to play in speeding the transition to a sustainable future, because a central aspect of sustainability is widespread behavior change. To date, however, most programs promoting sustainable behavior have featured information-intensive campaigns that make little use of psychological knowledge. Community-based social marketing is an attractive alternative approach in which promoters identify the activity to be promoted and the barriers to this activity and then design a strategy to overcome these barriers, using psychological knowledge regarding behavior change. The strategy is piloted to test its effectiveness and later evaluated when it is implemented on a broader scale. Unlike many information-intensive campaigns, community-based social marketing has been shown to have a much greater probability of promoting sustainable behavior. Two case studies are provided to illustrate the approach and its possible results. PMID- 10842435 TI - Flynn effects not genetic and unrelated to race differences. PMID- 10842436 TI - IQ gains and fluid g PMID- 10842437 TI - [Clinical epidemiology: a fundamental tool in clinical practice]. PMID- 10842438 TI - Death as a sentinel event: the mortality experience of Puerto Ricans in the United States. AB - The mortality data for 1996 and 1997 reveal that Puerto Ricans in the United States lag behind other Hispanics in terms of age-adjusted death rates. The better health status of Cubans in the United States can be explained by their immigration history and more favorable socio-economic conditions. The greater mortality risks of mainland Puerto Ricans compared to Mexican-Americans is more difficult to explain. While Puerto Ricans have more entitlements, higher incomes, and more education than their Mexican counterparts, the Mexicans have higher levels of employment and more stable families, indicators which apparently affect the relative risk of death. The data also show that, of all the Hispanic subgroups, the Puerto Ricans exhibit the most marked gender differentials, mainland Puerto Rican men being significantly more at risk than females. Finally, the data on infant mortality show that babies born to Puerto Rican women are less likely to survive their first year of life than infants born to women of other Hispanic origins. Each of these indicators suggests the greater vulnerability of mainland Puerto Ricans and confirms the disadvantaged status of this population vis-a-vis other groups. PMID- 10842439 TI - [A comparative study of indinavir versus ritonavir for assessing the development of opportunistic diseases in adult AIDS patients]. AB - This observational study compared the probability of developing the first opportunistic infection among AIDS adult patients attending the "Programa SIDA de San Juan" who received either indinavir plus zidovudine and lamivudine (n = 45) or ritonavir plus zidovudine and lamivudine (n = 16) between August 1, 1996 and July 31, 1997. No significant differences were observed with respect to appearance of an opportunistic infection, increase in CD4 levels and decrease in viral load between both groups during the study period. However, an increased probability of being free of opportunistic infection after 10 months was observed for the indinavir group (p > 0.05). In addition, the probability of changing or interrupting prescribed therapy was 2 times higher for patients under ritonavir (p < 0.05). These results suggest the need to confirm these findings in a larger group of patients in a controlled clinical trial and to assess the short-term and long-term effects of both therapies among Puerto Rican AIDS patients. PMID- 10842441 TI - Primary cardiac osteogenic sarcoma treated with heart transplantation. AB - Primary cardiac osteogenic sarcomas are rare malignancies. Less than 30 cases have been reported in the literature. Most of them have been found in the left side of the heart. We report a case of a primary osteogenic sarcoma of the left atrium in a 28 year old female. She underwent resection of the tumor and subsequently heart transplantation. To our knowledge this is the second patient with a primary cardiac osteosarcoma who underwent heart transplantation. PMID- 10842440 TI - [The prevalence of asthma and the utilization of medical services among those insured by a health-services company in Puerto Rico, 1996-1997]. AB - OBJECTIVE: To describe the prevalence and pattern of utilization of medical services in insured of SSS with a diagnosis of asthma during 1996 and 1997. METHODS: The medical claims of SSS insured whose main diagnosis was asthma (ICD-9 9 493-493.9) were selected for analysis. The prevalence and medical service utilization (medical visits, emergency and hospital admissions) were estimated. Differences in health service utilization by age group were analyzed by the Poisson model. RESULTS: The asthma prevalence was 14.5%, being larger in patients younger than 18 years of age and in females. 54.3% of the asthmatic patients visited medical offices and the larger proportion of users was observed in the younger group (< 18 years). However, the larger proportion of users of the emergency room was observed in the 18-44 age group, while the hospital admissions was larger in the 45-64 age group. More than half (56%) of the cost per service was attributed to hospital admissions while 31% was for pharmacy services. 65.9% of the insurers with asthma had prescriptions for short relief beta-antagonist. CONCLUSIONS: The prevalence of asthma in this study was high and similar to rates of the disease reported in Puerto Ricans residing in the U. S. and in other areas of the island. Similarly, the prevalence differed by age in the utilization of medical services as well as the high cost of hospital admissions. Prevalence studies using other sources as well as a standard definition of the condition may be helpful to confirm these results. PMID- 10842443 TI - Clostridial sepsis: is death avoidable? AB - Massive intravascular hemolysis is a rare yet often fatal complication of clostridial sepsis. The only chance for survival is an early diagnosis and prompt initiation of treatment. We report a rapidly fatal case who developed electrocardiographic changes of acute myocardial injury. Autopsy showed gas filled bubbles and cysts in the myocardium partially filled with sporulating bacilli with the morphology of clostridia. Gas filled bubbles were also present in the lungs, liver, kidneys and spleen. The gastric mucosa showed hemorrhagic and necrotizing changes, the probable site of entry of the infection. PMID- 10842442 TI - Prevalence of hepatitis C virus infection at three hemodialysis units in the western region of Puerto Rico. AB - OBJECTIVE: In the United States hepatitis C virus infection (HCV) affects approximately 20 percent of hemodialysis patients but its prevalence in Puerto Rico has not been established. We have sought to determine the prevalence of HCV infection in a homogeneous sample of patients on hemodialysis in the western region of Puerto Rico and to identify its risks factors. METHODS: All patients in the hemodialysis units of Aguadilla, Mayaguez and San German, during December 1997 to March 1998, completed a written questionnaire in which they were asked about transfusions, multiple sexual partners, i.v. drugs use, tattooing, occupation, imprisonment, organ transplantation and years on hemodialysis. Serum samples were analysed for HCV antibodies by an enzyme-linked immunoadsorbent assay (ELISA). Sera with positive results for HCV were subjected to a confirmatory test by the polymerase chain reaction (PCR). RESULTS: Thirteen of the 376 (3%) subjects had a positive ELISA (one patient died prior the confirmatory test with PCR). Six out of the twelve patients had a positive PCR. Two had been transfused. Three were illicit i.v. drug users and one had received a renal transplant. The liver biopsies in all patients showed chronic hepatitis and in two there was cirrhosis. CONCLUSIONS: Our prevalence was two percent. As reported elsewhere blood transfusion, organ transplantation and illicit i.v. drug abuse were the major risk factors for HCV infection in our patients. Nosocomial factors were irrelevant in the results. PMID- 10842444 TI - [The long-term psychological repercussions of sexual abuse in childhood: a pilot study in Puerto Rico]. PMID- 10842445 TI - Advances in the clinical assessment of dissociation: the SCID-D-R. AB - A comprehensive assessment of dissociative symptoms is recommended for effective treatment of trauma survivors. The author reviews the systematic detection of dissociative symptoms and disorders using the Structured Clinical Interview for DSM-IV Dissociative Disorders--Revised (SCID-D-R) in adolescents and adults (Steinberg, 1994b). Numerous investigations have reported good-to-excellent reliability and validity of the SCID-D-R, both in the United States and abroad. Clinical applications, including the SCID-D-R's utility for psychological and forensic evaluations, treatment planning, differential diagnosis, and evaluation of malingering, are reviewed. PMID- 10842446 TI - The differential diagnosis of epilepsy, pseudoseizures, dissociative identity disorder, and dissociative disorder not otherwise specified. AB - The authors review the co-occurrences of dissociative symptoms and disorders with epilepsy and pseudo-seizures and examine newer diagnostic instruments that assist in accurate diagnosis of persons with concomitant seizure behaviors and dissociative symptoms. They also review seizure behaviors and electroencephalographic findings in persons with dissociative identity disorder (DID) and dissociative disorder not otherwise specified (DDNOS) and dissociative symptoms in persons with epilepsy and with pseudoseizures. Dissociative symptoms in 15 patients with epilepsy and 15 with pseudo-seizures were examined using the Dissociative Experiences Scale (DES) and the Structured Clinical Interview for DSM-IV Dissociative Disorders (SCID-D). On the SCID-D, pseudo-seizure patients had significantly higher dissociative symptom scores than epileptic patients, but DES scores did not reliably distinguish epileptic and pseudo-seizure patients. Misdiagnosis of persons with seizures and dissociative symptoms can be avoided by careful adherence to DSM dissociative disorder criteria, the use of video-EEG monitoring, and systematic assessment of dissociative symptoms with the SCID-D. PMID- 10842447 TI - Preventing PTSD in trauma survivors. AB - This article highlights some of the core issues in the prevention of PTSD in trauma survivors. A review of some of the noteworthy research is presented along with new directions for research, training, education, and social planning efforts. PMID- 10842448 TI - The evolving standard of care in the practice of trauma and dissociative disorder therapy. AB - The standard of care for the delivery of mental health services has recently undergone a dramatic change spurred on by hundreds of lawsuits against therapists and by erupting controversies in the science of trauma and dissociative disorders. The nature of these changes and the forces that created them are discussed. Risk management suggestions are provided. PMID- 10842449 TI - Course of illness following specialized inpatient treatment for women with trauma related psychopathology. AB - The authors report findings from repeated assessments of 101 women who received intensive inpatient treatment focused on trauma-related disorders. All patients were assessed at admission and discharge, and half were reassessed at 1-year follow-up. Measures included the Global Assessment Scale, Brief Psychiatric Rating Scale, Brief Symptom Inventory, Role Functioning Scales, Follow-up Adjustment Scale, and Client Satisfaction Questionnaire. As a group, patients improved substantially during hospitalization and maintained gains at follow-up, although they showed a trend toward relapse in some symptom areas. However, a substantial minority of patients continued to show severe impairment at follow up. In light of continual need for changes in clinical programs in the face of declining reimbursement, there is a pressing need for future outcome research on specialized trauma programs to relate subsequent clinical course to specific treatment interventions. PMID- 10842450 TI - Pediatric and neonatal intensive care hospitalization as traumatic stressor: implications for intervention. AB - A substantial literature documents the psychological, medical, and economic benefits of attending to the emotional needs of parents whose children are hospitalized on the pediatric intensive care unit (PICU) or neonatal intensive care unit (NICU). Synthesizing the PICU/NICU literature with current findings from the field of psycho-traumatology, the author addresses PICU/NICU hospitalization as a potential traumatic stressor for families of hospitalized children and considers that intensive care clinical staff are vulnerable to symptoms of primary and secondary traumatic stress as well. Guidelines are offered for recognizing, managing, and increasing resiliency to the symptoms of normal traumatic stress responses in both families and intensive care staff. PMID- 10842451 TI - Lives through time: an ideographic approach to the study of resilience. AB - The authors describe and illustrate an ideographic method for the study of resilience. The method assumes that resilience is an unfolding and dynamic process in which the individual and the social environment interact to produce a life course over time. Using retrospective and prospective data, the authors present a case study of a profoundly troubled boy, first seen as a preschooler and then followed up in early adulthood with a battery of clinical interviews. The methodology presented provides a way to study resilience as a dynamic process and to reveal and make sense of counterintuitive findings. In contrast to historical or epidemiological methods, the approach also permits exploration of the role of intrapsychic processes in facilitating change. PMID- 10842452 TI - Graduation and other psychiatric emergencies. PMID- 10842453 TI - Outbreak of Legionnaires' disease associated with aquarium in Australia. PMID- 10842454 TI - Baby dies of Salmonella poona infection linked to pet reptile. PMID- 10842455 TI - Characterization and variability of endotoxin and 3-hydroxy fatty acids in an office building during a particle intervention study. AB - Air and dust samples were collected on two floors of an office building during a double-blind particle intervention study to examine spatial and temporal variability of airborne endotoxin over a period of weeks, and to characterize endotoxin activity and lipopolysaccharide (LPS) content in carpet and chair dust. Air samples were collected on multiple days within and across weeks. Dust samples were collected from carpets and chairs one day per week for three weeks. Endotoxin was measured using a Limulus assay. Dust samples were analyzed for LPS by determination of 3-hydroxy fatty acids (3-OHFAs) using gas chromatography-mass spectrometry. The geometric mean (geometric standard deviation) for 96 indoor air samples was 0.24 (1.6) EU/m3. Significant within-floor spatial variation of airborne endotoxin was found (P < 0.0001, n = 80). Temporal variability of airborne endotoxin was not significant across weeks. Mean (+/- SD) endotoxin levels in carpet dust (59 +/- 9.3 EU/mg dust, n = 12) and in chair dust (38 +/- 7.7 EU/mg dust, n = 10) were significantly different (P < 0.001). Carbon chain length-dependent differences in 3-OHFA levels by dust source and floor were found. Enhanced air filtration did not significantly affect airborne endotoxin (P = 0.62); however, total dust mass and total endotoxin in carpet dust samples increased significantly after enhanced surface cleaning (P < 0.01). These findings suggest that spatial variability, dust source, and surface cleaning may influence building occupant exposures to endotoxin. PMID- 10842456 TI - Determination of microbial colonisation in water-damaged buildings using chemical marker analysis by gas chromatography-mass spectrometry. AB - Gas chromatography-mass spectrometry was used to determine the microbial contents of building materials subjected to water damage in a laboratory experiment and of materials collected from houses affected by water during the flood in Klodzko in south-western Poland, July 1997. The samples were examined for 3-hydroxy fatty acids, markers of bacterial endotoxin, and ergosterol, marker of fungal biomass. The amounts of both 3-hydroxy fatty acids and ergosterol were higher in materials that had been exposed to water than in unexposed ones. All markers were stable in the building materials for at least 6 weeks at room temperature and could thus be used to reveal microbial contamination even when cultivation results for bacteria and fungi were negative. Direct measurement of 3-hydroxy fatty acids and ergosterol in human environments could be a useful method, e.g. in monitoring indoor air as regards presence of potentially harmful microorganisms and microbial constituents. PMID- 10842457 TI - Effect of ventilation and filtration on submicrometer particles in an indoor environment. AB - The effect of filtration and ventilation on reduction of submicrometer particle concentration indoors was investigated in an office building. The air-handling system consisting of dry media filters and an air-conditioning unit, reduced particle concentration levels by 34%. The characteristics of indoor airborne particles were dominated by, and followed the pattern of, outdoor air, with vehicle combustion aerosols as the main pollutant. The ratio indoor/outdoor particle concentration varied between 14 and 26% for different sub-zones. The presence of significant source of particles indoors was not observed. A simple mathematical model predicting evolution of particles indoors is presented. The model, based on a particle number balance equation, was validated with experimental data and showed very good agreement between predicted and measured parameters. PMID- 10842458 TI - Introduction of a sink-diffusion model to describe the interaction between volatile organic compounds (VOCs) and material surfaces. AB - A sink-diffusion model to describe the interaction between material surfaces and volatile organic compounds (VOCs) in indoor air has been introduced. The model is based on adsorption/desorption on the material surfaces and diffusion into the materials. Test chamber experiments with exposure of nylon carpet and polyvinyl chloride (PVC) covering against alpha-pinene and toluene were used to validate the model and to make comparisons with a sink model based on the Langmuir adsorption isotherm. The results showed that the sink-diffusion model gave a better description of the desorption curve than the Langmuir model. The model predictions improved with increasing sorption effect. The Langmuir model gave good predictions of relative weak sorption effects, whereas the sink-diffusion model improved the predictions for stronger sorption effects. In this case, nylon carpet showed substantial stronger sorption than PVC covering and alpha-pinene showed stronger sorption than toluene. Controlled field experiments with combinations of building materials and a mixture of VOCs, encountered in real indoor environments, are needed to further validate the sink-diffusion model. PMID- 10842459 TI - The biofiltration of indoor air: implications for air quality. AB - An alternative method of maintaining indoor air quality may be through the biofiltration of air recirculating within the structure rather than the traditional approach of ventilation. This approach is currently being investigated. Prior to its acceptance for dealing with volatile organic compounds (VOCs) and CO2, efforts were made to determine whether the incorporation of this amount of biomass into the indoor space can have an (negative) impact on indoor air quality. A relatively large ecologically complex biofilter composed of a ca. 10 m2 bioscrubber, 30 m2 of plantings and a 3,500 litre aquarium were established in a 160 m2 'airtight' room in a recently constructed office building in downtown Toronto. This space maintained ca. 0.2 air changes per hour (ACH) compared to the 15 to 20 ACH (with a 30% refresh rate) of other spaces in the same building. Air quality parameters of concern were total VOCs (TVOCs), formaldehyde and aerial spore counts. TVOC and formaldehyde levels in the biofilter room were the same or significantly less than other spaces in the building despite a much slower refresh rate. Aerial spore levels were slightly higher than other indoor spaces but were well within reported values for 'healthy' indoor spaces. Levels appeared to be dependent on horticultural management practices within the space. Most genera of fungal spores present were common indoors and the other genera were associated with living or dead plant material or soil. From these results, the incorporation of a large amount of biomass associated with indoor biofilters does not in itself lower indoor air quality. PMID- 10842460 TI - Hygroscopic fine mode particle deposition on electronic circuits and resulting degradation of circuit performance: an experimental study. AB - A portion of electronic equipment failures is a consequence of particle deposition on electronic circuits in normal indoor environments. Deposited hygroscopic particles reduce the electrical isolation (EI) between conductors. In laboratory experiments, we investigated the mechanisms, locations, and effects of particle deposition on electronic circuits with surface mounted chips (SMCs) and also on small television sets. One set of electronics was exposed for 281 h to an unusually high concentration of artificially-generated ammonium sulfate particles while a second set (experimental controls) was exposed to normal indoor particles. The particle mass concentration in the high-exposure chamber was 500 times higher than normal. Television reliability was observed and the changes in EI between adjacent legs of SMCs were measured. The experiments demonstrate the strong influence of electrostatic forces on the locations and rates of particle deposition. Although televisions did not fail after exposure to concentrated aerosols, the EI between adjacent legs of the SMCs was, in many cases, greatly diminished. Relative humidity had a very strong influence on the magnitude of EI. A qualitative explanation of the mechanisms of particle deposition and circuit degradation is proposed, including the role of fibers. Finally, a potential method to reduce particle deposition on electronic components is discussed. PMID- 10842461 TI - Improved health after intervention in a school with moisture problems. AB - In a school with floor moisture problems, the personnel had complaints consistent with the sick-building syndrome (SBS). Interventive measures including the laying of a ventilated floor were undertaken to eliminate the emissions. To examine if the intervention resulted in positive health effects, 34 personnel and 336 pupils were interviewed just before the intervention and also 7 months after. Also were interviewed 21 personnel and 224 pupils at an adjacent school serving as a control. Compared with the control school, the problem school showed more complaints, more general symptoms and more symptoms from the eyes, airways and skin, both among the personnel and the pupils. In the post-intervention examinations, the excess of symptoms among the personnel had almost disappeared. Among the pupils, the frequency of eye irritation was reduced but a general improvement of the other symptoms was not as obvious. However, after adjustment for a recent common cold, atopy and stress among the pupils, only one symptom ("stuffy nose") remained significantly elevated. In conclusion, the intervention was followed by positive health effects, supporting the hypothesis that emissions from building material had contributed to the excess of symptoms. A recent common cold was highly related to the symptoms and should be considered in future SBS studies. PMID- 10842462 TI - Histopathological and immunohistochemical studies on cetaceans found stranded on the coast of Italy between 1990 and 1997. AB - Detailed histopathological and immunohistochemical studies were carried out on 24 cetaceans, 17 of which were striped dolphins (Stenella coeruleoalba), four bottlenose dolphins (Tursiops truncatus) and three Risso's dolphins (Grampus griseus), all found stranded on the Italian coast between 1990 and 1997. The most frequently detected lesions were chronic pneumonia (73.7% of the examined lungs), focal chronic pancreatitis (71.4%), non-purulent encephalitis (50%), chronic hepatitis (42.1%), and chronic focal interstitial nephritis (31.2%). The skin and the subcutaneous panniculus were often (33.3%) affected by parasitic diseases (Phyllobothrium delphini and Pennella sp.). An appreciable percentage of animals showed lymphoid depletion in their lymphatic organs (47.2%), as well as a high rate of parasitic infestations in their alimentary tracts (25%). PMID- 10842463 TI - Evaluation of cellular and biochemical parameters of blood and peritoneal fluid following exploratory laparotomy in the goat. AB - To evaluate the effects of exploratory laparotomy on cellular and biochemical parameters of blood and peritoneal fluid, an experiment was conducted using 10 Iranian cross-bred male goats. Approximately 10 ml of blood and 1-1.5 ml of peritoneal fluid were collected from all animals prior to operation for estimation of control values. Exploratory laparotomy was performed under local analgesia. Blood and peritoneal fluid samples were collected at 24, 48, 72 and 96 h after exploratory laparotomy. The results revealed that after exploratory laparotomy, the number of white blood cells and the percentage and absolute number of neutrophils and band neutrophils significantly increased (P < 0.05). However, the percentage of lymphocytes decreased significantly (P < 0.05). The concentrations of blood urea nitrogen significantly increased (P < 0.05). Furthermore, following the operation, the percentage and absolute number of neutrophils in the peritoneal fluid significantly increased (P < 0.05). In contrast, the percentage of lymphocytes in the peritoneal fluid decreased significantly (P < 0.05). The concentration of protein in the peritoneal fluid increased significantly (P < 0.05). PMID- 10842464 TI - Brain, liver and plasma unsaturated aldehydes in nutritional encephalomalacia of chicks. AB - Vitamin E deficiency and linoleic acid-feeding lead to nutritional encephalomalacia (NE) in chicks, affecting the cerebellum exclusively. The relevance of lipid peroxidation (LPO) products to the pathogenesis of the disease was studied. Laying hens received a diet low in vitamin E. Resulting chicks were assigned to four groups fed either with linoleic (C18: 2n-6) or linolenic (C18: 3n-3) acid together with 1 or 50 p.p.m. vitamin E. Nine days post-hatching NE occurred in the vitamin E-deficient group fed linoleic acid. With each chick showing NE, a healthy one from all four groups was killed. Unsaturated aldehydes were determined in plasma, liver, cerebrum and cerebellum. Results underlined that the type of dietary fat is decisive for the aldehyde pattern. In the liver of linoleic acid-fed animals total aldehydes were increased. Diseased animals had increased aldehydes stemming from n-3 fatty acids. In plasma, vitamin E deficiency led to higher malondialdehyde and OH-nonenal concentrations. In brain, neither vitamin E deficiency nor NE were accompanied by increased aldehyde concentrations. In consequence a direct role of unsaturated aldehydes for the development of NE in the cerebellum is not probable. PMID- 10842466 TI - Epidemiology of infectious keratoconjunctivitis in cattle in south-east Ethiopia. AB - The aim of the study was to determine the prevalence of infectious keratoconjunctivitis on cross-breed dairy farms and to evaluate factors that influence its distribution. A total of 11 dairy farms and one cattle cross breeding farm in different parts of Arsi region, south-east Ethiopia were clinically and bacteriologically investigated from February 1989 to December 1990. Out of 5221 local zebu and cross-breed dairy animals, 110 were found to have different degrees of infectious bovine keratoconjunctivitis. Of these, 94 (85.5%) were unilateral and 16 (14.5%) were bilateral. The prevalence of the disease was significantly (P < 0.001) influenced by age and breed; high exotic blood level and younger age groups were mainly affected. However, severity of the disease was not significantly associated with age (P > 0.750). Moraxella bovis was isolated from 88 (80%) of the 110 bacteriologically positive cases. Of these isolates, 61 (69%) were haemolytic and 27 (31%) were non-haemolytic strains. Infection with M. bovis was unilateral. Concurrent infections with M. bovis included Actinomyces pyogenes 23 (21%), Staphylococcus aureus 17 (15%), Pasteurella haemolytica nine (8%), Escherichia coli eight (7%) and Proteus spp. four (3.6%). PMID- 10842465 TI - Tissue distribution of oxacillin and ampicillin in the isolated perfused bovine udder. AB - In vivo, tissue distribution of intra-mammarily administered antibiotics is mostly only assessed by sampling milk and blood. Therefore, the described study analysed whether measurement of tissue concentrations makes sense in vitro instead. Isolated bovine udders were perfused with gassed and warmed Tyrode solution. To four front and rear quarters each, 1000 mg oxacillin in 7.5 ml vehicle was administered intracisternally, completely formulated as sodium monohydrate in two lactation ointments (with or without sodium dodecylsulphate) or 80% as benzathine salt in a dry-off ointment. Over 3 h, perfusate and glandular tissue from different locations were sampled and analysed by high pressure liquid chromatography. With increasing vertical distance to the teat base, the tissue concentration of antibiotics decreased. With the lactation ointment containing sodium dodecylsulphate, lower oxacillin concentrations were reached in glandular tissue and lymph nodes compared to those without. The ointments led to a higher recovery of oxacillin in glandular tissue than in perfusate. Aluminium monostearate in the dry-off ointment caused an even poorer absorption of oxacillin into perfusate. The isolated perfused bovine udder is suitable to study the tissue distribution of antibiotics, since the results were mainly comparable with the few existing in vivo studies and show the influence of different formulations. PMID- 10842468 TI - Age-associated changes in the immune system of German shepherd dogs. AB - In order to look into the ageing of the canine immune system we investigated age related changes and associated gender-related differences in parameters of innate and acquired immunity in German Shepherd dogs. We obtained the following findings: white blood cell counts, peripheral blood lymphocytes, lymphocyte proliferative activity and interleukin-2 (IL-2) serum concentrations were significantly lower in the group of old animals, whereas the concentrations of gamma-globulins and the functional activity of the complement system were significantly higher in the elderly. Phagocytic and bactericidal activity of polymorphonuclear cells, as well as their 'killing function,' the serum cytokine like activities of tumour necrosis factor-alpha and the plasma concentrations of immunoglobulin G, as well as of alpha- and beta-globulins, were not significantly affected by age, whereas natural killer-cell activity and the serum cytokine-like activities of IL-1 were significantly higher only in the group of female old animals. With regard to gender-related differences, lymphocyte proliferative activities as well as plasma concentrations of alpha-globulin were significantly higher in the group of female animals, whereas the absolute numbers of segmented neutrophils were significantly lower. Species analogies with regard to ageing as presumed to exist between man and laboratory rodents also seem to be applicable to the dog. The observed age-related changes in the canine immune system are probably among the main causes for the multimorbidity of old age, affecting life expectancy and mortality in the dog and should be recognized and considered by the attending veterinarian. PMID- 10842467 TI - Detomidine-diazepam-ketamine anaesthesia in buffalo (Bubalus bubalis) calves. AB - Eight buffalo calves (8-12 months, 70-100 kg) were randomly assigned to two groups of four animals each. Animals of group I were given detomidine (100 micrograms/kg), whereas animals of group II received a mixture of detomidine (100 micrograms/kg), diazepam (100 micrograms/kg) and ketamine (3 mg/kg) (DDK) intravenously. Various clinical parameters, such as weak time, down time, pedal and pinprick reflexes, muscle relaxation and extent of sedation, as well as heart and respiratory rates and electrocardiograms were measured before (time 0) and 15, 30, 45, 60, 75 and 90 min after treatment. In all the animals of group II (DDK), the pedal reflex was completely abolished (score: 3.00 +/- 0.00) within 5 min, the pinprick response was either very weak or it was completely abolished at this interval. Muscle relaxation and sedation were excellent within 5 min of DDK administration. The depth of sedation and analgesia was maximum from 5 to 15 min postinjection. Detomidine alone, however, failed to produce appropriate depression of the pedal and pinprick reflexes, sedation was mild and muscle relaxation was inadequate. Heart rate showed a significant (P < 0.05) decrease in group I, but the decrease was non-significant in group II. A more pronounced increase in respiratory rate was observed in group I as compared to group II. Animals of both groups recovered within 90 min without any complication. Minimal changes in the cardiovascular system in the group given the DDK combination were an advantage over the group given detomidine. The results indicated that DDK combination is safe and suitable for 15 min of anaesthesia with excellent muscle relaxation and has only limited cardiorespiratory effects in buffaloes. PMID- 10842469 TI - [Tips for evaluating airplane flight fitness. Doctor, may I fly?]. PMID- 10842470 TI - [Mechanical barrier against pollen. Nose ointment to control hay fever?. Interview by Dipl.-Biol. Waltraud Paukstadt]. PMID- 10842471 TI - [Vertigo as a diagnostic challenge. When everything revolves around your patient]. PMID- 10842472 TI - [Diagnosis of Lyme borreliosis. How to corroborate suspected borreliosis]. AB - Lyme borreliosis is a zoonosis transmitted via lxodes ticks. The causal agent- the spirochete Borrelia burgdorferi sensu lato--triggers a multisystem disease that manifests in particular in the skin, heart, nervous system and joints. Lyme borreliosis is the most common tick-borne infectious disease in the Northern hemisphere. In Europe the Lyme disease spirochetes are heterogeneous and comprise three species that infect humans: Borrelia burgdorferi sensu stricto, B. garinii and B. afzelii. The microbiological diagnosis is based primarily on the detection of antibodies, and secondarily on the detection of the pathogen. Suitable material for the detection of the pathogen are various body fluids (cerebrospinal fluid, joint fluid) and biopsy material (in particular skin). Antibodies are usually detected in the serum. Negative serology does not exclude an early manifestation, and a positive finding is no proof of a clinically manifest infection--it may simply reflect an earlier Lyme infection. PMID- 10842473 TI - [Therapy of Lyme borreliosis. Stage-appropriate antibiotic selection and dosage]. AB - When there are good reasons for suspecting Lyme borreliosis, treatment with an antibiotic is mandatory. If the clinical picture clearly shows erythema migrans, antibiotic treatment must be instituted also in the absence of a bacteriological diagnosis. Later manifestations require longer and higher-dose antibiotic treatment. Except acrodermatitis stage II and III manifestations should be treated intravenously. The success of treatment cannot be monitored serologically. A European vaccine is currently being developed. PMID- 10842474 TI - [Pollen allergy. A plea for desensitization]. PMID- 10842475 TI - [Therapy of hypothyroidism. T4 or better T3/T4?]. PMID- 10842476 TI - [Alarm from the diabetic kidney. Diagnostic series: Microalbuminuria]. PMID- 10842477 TI - [Risk of thrombosis in malignant diseases. Thrombosis ABC, 15]. PMID- 10842478 TI - [Exanthematous infections in childhood. 1: Rubella, measles, Pfeiffer's glandular fever]. PMID- 10842480 TI - [In Process Citation] PMID- 10842479 TI - [Young women with pulsating tumor. Horseshoe kidney]. PMID- 10842481 TI - [Sex offenders: dangerousness is perhaps overestimated by psychiatrists. Interview with Drl Herbert Steinbock of the Haar district hospital. Interview by Dr. Anita Schweiger]. PMID- 10842483 TI - Development of the cavernous sinus in the fetal period: a morphological study. AB - The development and morphological structure of the lateral sellar compartment (LSC), an interdural space containing the cavernous sinus (CS), cranial nerves, and internal carotid artery (ICA), was investigated by histological examination of sections of the LSC and cerebral venograms from human fetal specimens. Twenty eight LSC coronal sections were obtained from 14 fetuses of 13-32 weeks' gestation. Venograms of 11 other fetuses of 13-32 weeks' gestation were studied to observe changes in venous drainage. The CS appeared as a collection of small venous canals with an endothelial layer. These venous canals gradually became much larger through expansion and unification. The CS and basilar venous plexus were demonstrated as a faint cluster of small vessels on venograms obtained after 13 weeks' gestation. The dura mater increased in thickness and collagen fiber networks developed around all the components in the LSC after 23 weeks' gestation. The LSC lateral wall could not be histologically differentiated as separate multiple layers. Branching and joining of the cranial nerve fascicles were completed with the envelopment of collagen fibers after 23 weeks' gestation. The ICA at 13-15 weeks' gestation ran straight within the LSC, becoming tortuous before birth. CS formation occurs through the development of venous canals without smooth muscle layers, followed by web-formation by collagen fibers in the mesenchymal interstices. LSC formation, including the dense dura mater and an internal structure like that seen in the adult, is largely completed before birth. PMID- 10842482 TI - Significance of shock in head-injured patients with skull fracture. AB - The clinical differences between patients with skull base and convexity fractures were retrospectively investigated in 324 patients, of whom 110 had suffered head injury resulting in skull fracture. These 110 patients were divided into the skull base and convexity groups. There were no significant differences between the groups with respect to sex, age, Glasgow Coma Scales, injury severity scores, pupil abnormalities, and outcomes. Automobile collisions were the most common causes in the skull base group, and falls in the convexity group. Traumatic Coma Data Bank diffuse 1 type injuries were more frequent in the skull base group and evacuated masses were more frequent in the convexity group. Multiple injuries, shock on admission, lower hemoglobin concentrations, and increased transfusion requirements were evident in the skull base group. Controlling for shock, the outcomes in the skull base group were favorable. Convexity fractures were usually associated with isolated severe head injuries and require brain protection therapy. Skull base fractures were caused by a significant force distributed over a large area of the body with a tendency to induce shock, and require a multidisciplinary approach to treatment. PMID- 10842484 TI - Postoperative changes in paraspinal muscle thickness after various lumbar back surgery procedures. AB - Damage to the paraspinal muscle after various lumbar back surgery procedures was evaluated by measuring the paraspinal muscle thickness preoperatively and postoperatively in 89 patients, 61 males and 28 females. There were 42 single interlaminar level procedures (SL group), 13 multiple interlaminar level procedures (ML group), and 34 posterolateral fusion procedures (PLF group). Changes in paraspinal muscle thickness were evaluated at more than 10 months after surgery, because muscle swelling continued as long as 10 months after surgery, and then reduced as the edema subsided. Postoperative serum creatine phosphokinase (CPK) level on postoperative day 2 was also measured. The decrease of paraspinal muscle thickness was significantly larger in the PLF group than in the SL group (-12.9% and -2.7%, respectively, p < 0.02). There was no significant correlation between postoperative serum CPK level and decrease of paraspinal muscle thickness. However, the postoperative elevation of serum CPK level was significantly higher in the PLF group than in the SL and ML groups (979 +/- 114 vs. 292 +/- 45 and 410 +/- 44 IU/l, respectively, p < 0.001). In conclusion, posterolateral fusion is the most invasive procedure of the paraspinal muscles in various lumbar back surgery procedures. Paraspinal muscle damage during lumbar back surgery may be one of the most important factors that causes atrophy of the muscles. PMID- 10842485 TI - Traumatic basal subarachnoid hemorrhage due to rupture of the posterior inferior cerebellar artery--case report. AB - A 20-year-old male presented with traumatic basal subarachnoid hemorrhage after being involved in a fight. Antemortem clinical examinations could not exclude the possibility of rupture of abnormal blood vessels because of the absence of external injuries. Careful postmortem examination of the head and neck regions and histological examination of the intracranial arteries demonstrated traumatic rupture of the left posterior inferior cerebellar artery due to a fist blow to the jaw. This case indicates the need for careful autopsy examination for the differentiation of traumatic and non-traumatic basal subarachnoid hemorrhages. PMID- 10842486 TI - Delayed cerebrospinal fluid rhinorrhea seven months after transsphenoidal surgery for pituitary adenoma--case report. AB - A 51-year-old female had undergone transsphenoidal surgery for pituitary adenoma producing growth hormone. Cerebrospinal fluid (CSF) leakage occurred during surgery. The sella turcica and sphenoid sinus were packed with abdominal fat and fibrin glue, buttressing the closure with a fragment of sphenoid bone. No CSF rhinorrhea occurred postoperatively. Severe meningitis developed 7 months later. CSF rhinorrhea occurred 10 days after readmission. Exploration through the transsphenoidal approach identified a small hole at the floor of the sella and CSF leaking into the sphenoid sinus through the hole. The CSF leakage stopped after the second surgery. Delayed CSF rhinorrhea without bromocriptine administration is very rare. The cause of delayed CSF rhinorrhea remains unclear. CSF rhinorrhea should be suspected if meningitis develops even months after transsphenoidal surgery. PMID- 10842487 TI - Spontaneous occlusion of ruptured vertebral artery dissection at the extradural fenestration associated with extradural origin of the posterior inferior cerebellar artery--case report. AB - A 69-year-old female suffered from sudden onset of severe headache. Computed tomography showed subarachnoid hemorrhage primarily located in the posterior fossa. Initial angiography demonstrated a fenestration of the vertebral artery and an extracranial origin of the posterior inferior cerebellar artery. However, no bleeding points could be clearly detected. The operative findings revealed a massive clot in subarachnoid space, but no bleeding point. Serial angiography demonstrated dissection in one of the limbs of the fenestrated vertebral artery on the 25th day after the onset. On the 100th day, the lesion was spontaneously occluded. The patient is presently doing well at 8 years after surgery. PMID- 10842488 TI - Posterior fossa arteriovenous malformation associated with persistent primitive trigeminal artery--case report. AB - A 21-year-old female presented with an unusual case of posterior fossa arteriovenous malformation (AVM) associated with ipsilateral persistent primitive trigeminal artery (PPTA), manifesting as intraparenchymal hemorrhage involving both the brain stem and the left cerebellar hemisphere. The presenting symptoms were compatible with Wallenberg's syndrome and Foville's syndrome on the left side. She was initially treated conservatively, and subsequently with transarterial embolization followed by stereotactic radiosurgery. This case combined the rare association of posterior fossa AVM and PPTA, with the clinical presentation of intraparenchymal hemorrhage causing both Wallenberg's syndrome and Foville's syndrome. PMID- 10842489 TI - Myelosarcoma of the cavernous sinus in a nonleukemic patient--case report. AB - Myelosarcomas are rare, solid extramedullary tumors composed of granulocyte precursors in most cases associated with leukemia. A 38-year-old nonleukemic female presented with a myelosarcoma of the cavernous sinus. After surgical removal of the cavernous sinus tumor she was treated by chemotherapy and whole brain radiation therapy. Despite this aggressive therapy, she died 4 months after surgical intervention. PMID- 10842490 TI - Chiari malformation and syringomyelia in monozygotic twins: birth injury as a possible cause of syringomyelia--case report. AB - A 26-year-old female, the elder of monozygotic twins, presented with slow progressive numbness and pain in her left arm. Magnetic resonance (MR) imaging showed syringomyelia with Chiari malformation. The patient's birth had been difficult with prolonged delivery time, breech delivery, and neonatal asphyxia. MR imaging of the patient's twin sister showed mild tonsillar ectopia, but absence of syringomyelia. This younger sister was born without problems. The patient underwent syringosubarachnoid shunt at the C5-6 level. The syrinx was collapsed promptly, and her symptoms disappeared. This case of syringomyelia with Chiari malformation in one of twins suggests that birth injury is likely to be a cause of the pathogenesis of syringomyelia. PMID- 10842491 TI - Isolated dilation of the trigono-inferior horn--four case reports. AB - Four patients presented with isolated dilation of the trigono-inferior horn associated with either mass lesion at the trigone of the lateral ventricle or with shunt over-drainage. We investigated clinical symptoms, course, and neuroradiological findings of these cases. The pressure of the isolated ventricle was measured or estimated at surgery in all cases. The common symptoms were recent memory disturbance and contralateral homonymous hemianopia. Contralateral hemiparesis was observed occasionally. Rapid deterioration of the isolation caused uncal herniation in one case. Comma-shaped dilation of the inferior horn was observed in all cases. Midline shift was not conspicuous except in one case. Intraventricular pressure at surgery was 18 cmH2O, 35 cmH2O, 3 cmH2O, and within normal range. These cases had very similar clinical symptoms and neuroradiological findings. The pathophysiology of isolation suggested three types of isolation (high-, normal-, and low-pressure isolation), depending on the pressure of the isolated ventricle. The isolation of trigono-inferior horn is an important clinical entity as it may cause uncal herniation in patients with high pressure lesions. PMID- 10842492 TI - Make the patient a friend for life. PMID- 10842493 TI - Referral. DOT-COM. PMID- 10842494 TI - Teachers still needed. PMID- 10842495 TI - Starting out. PMID- 10842496 TI - What is your practice worth? AB - Time has proven that expert valuation of dental practices combines both art and science. The valuation results will vary according to the methods used, and, frequently, the most difficult task is to reconcile the various values derived through different acceptable valuation techniques. PMID- 10842497 TI - Downsize your debt. PMID- 10842498 TI - Are husbands a barrier to women's family planning use? The case of Morocco. AB - Little is known about men's role in the adoption of family planning. Recent studies suggest that men may be a barrier to women's use of family planning. However, it is not clear whether husbands represent a true or a perceived barrier. Using the 1992 Morocco Demographic and Health Survey data, this study examines (1) whether women and men report concordant fertility desires, discussions, and contraceptive use; (2) the accuracy of women's perceptions of their husbands' fertility desires; and (3) whether husbands are a barrier to women's family planning use. The results demonstrate that, controlling for women's own fertility desires, husbands' true fertility desires are associated with family planning use. Likewise, women who perceive their husbands to want fewer children than they want are more likely to use family planning. Future fertility and family planning programs need to include men to reduce their role as both perceived and true barriers to family planning use. PMID- 10842499 TI - Ethnicity and sexual behavior in Ghana. AB - Using data from the 1993 Ghana Demographic and Health Survey, this study explores the relationship between ethnicity and sexual behavior: having sex before age 17 and premarital sexual experience. All ethnic groups show substantial sexual experience before age 17 and premarital sexual engagement. Logistic regression analyses reveal that in general ethnicity influences the behaviors studied, especially for ever-married women. The data suggest that groups that practice matrilineal and patrilineal systems show differences in the likelihood of having sex before age 17. Contrary to expectation, there is an inverse relationship between education and sexual experience before age 17. The findings highlight the importance of group-specific programs in Africa. PMID- 10842500 TI - The secondary sex ratio in Israel: 1980-1995. AB - This study deals with models and hypotheses that attempt to explain the underlying mechanisms determining the sex ratios at birth in human populations. Since the factors responsible are still questionable and research results are contradictory, we examine data available in Israel on the sex ratios at birth among two different sociodemographic groups, Jews and Moslems. Results suggest a difference between Jewish and Moslem patterns of secondary sex ratios with respect to parental age, education, and birth number. The difference may be described as a more regular and, by existing models, a more predictable pattern of secondary sex ratio among Moslems than among Jews. The possibility that Jewish religious laws play a role in this difference is discussed. PMID- 10842501 TI - Premarital childbearing in northwest Kenya: challenging the concept of illegitimacy. AB - A number of African countries, including Kenya, have experienced a marked rise in births among unmarried women. In Western countries, reproduction outside of marriage is assumed to be illegitimate and a social problem. One hypothesis used to explain the increasing incidence of premarital fertility in Africa is a breakdown of traditional social controls by the extended family over the sexual behavior of adolescents. A competing hypothesis suggests that unmarried women use sexual relations to achieve goals such as marriage. Among Turkana pastoralists of northwest Kenya, we find a pattern of premarital birth that fits either hypothesis only loosely. Premarital fertility among the Turkana is both widespread and culturally acceptable, with over 30 per cent of women having at least one child prior to marriage. Although women with premarital births initiate childbearing on average one year earlier than women with only marital births, women's marital status does not influence the length of the interval between first and second births. Marriage among the Turkana is not a social trigger for the onset and continuation of reproduction or a means to legitimate reproduction. Marital status of the parents simply determines the custody of a child. In a premarital birth, the father pays a set fee to the mother's family, and the custody of the child remains permanently with the mother's family. If the parents later marry, the father must purchase custody of the child by another fee at that time. Since the Turkana have experienced few effects of modernization, the existence of such a practice suggests that cultural factors must be taken into account before assessing premarital fertility in Africa as a social problem. PMID- 10842502 TI - Structural determinants of the abortion rate: a cross-societal analysis. AB - Data for a sample of 89 countries are used to examine societal-level predictors of the legal status of abortion and its incidence as indicated by abortion rates. Measures of socioeconomic development, women's labor force participation, and dominant religion are considered as predictors of abortion. Logistic regression analysis reveals that socioeconomic development has a positive effect on the probability of abortion being legal. Both a greater dominance of Catholicism and Islam in a society reduce the probability that abortion is legal. Results of tobit analyses show that development has curvilinear effects on abortion rates, with lower rates of abortion at both the lower and higher ends of development. Findings also indicate a positive effect of female labor force participation on the abortion rate that initially grows stronger with increases in socioeconomic development, but weakens with further increases. Finally, a greater proportion of Catholics in a population lowers the abortion rate. PMID- 10842503 TI - Reproductive change in Zimbabwe and Kenya: the role of the proximate determinants in recent fertility trends. AB - This study examines trends in proximate determinants of fertility in Zimbabwe and Kenya. Findings from the four Demographic and Health Surveys conducted in these countries show that the dramatic fall in fertility in these countries is consistent with the underlying trends in the most important proximate determinants of fertility. In Zimbabwe, contraceptive use far exceeds other proximate determinants in influencing fertility levels and trends. The results show that the fertility inhibiting effects of contraception are more important than the effects of postpartum infecundability, marriage patterns, or sterility. The results also show that contraceptive use has its greatest suppressing effects in the middle and younger age groups. However, in Kenya, the dominant fertility inhibiting effect is postpartum infecundability, with contraception coming in second. PMID- 10842504 TI - Determinants of the risk and timing of alcohol and illicit drug use onset among natives and non-natives: similarities and differences in family attachment processes. AB - This study examines data from Native Ontario reserve residents (Embree, 1993) and a sample from the Ontario Health Survey Supplement (1990-91) in order to compare and contrast the importance of family attributes such as parent-child attachment to Native and Non-native patterns of drug and alcohol use onset. Proportional Hazards modeling (Cox, 1972) was employed to identify factors associated with the risk and timing of onset of alcohol and illicit drugs for both cultural groups. For both Natives and Non-natives alike, and considering both drinking and drug use onset together, age cohort predominates as a risk factor, with youngest groups at greatest risk, and especially in the case of drug use other than alcohol. For the model of drug use timing, age of alcohol use onset is the second best predictor for Natives, although its effect is still apparent, albeit weaker, in the case of Non-natives. As for family characteristics, a number of factors emerge as determinants of risk and depend, in part, on the cultural group and the substance under consideration. Consistent with attachment theory's prediction about the universal applicability of the need for close parent-child relations (Bowlby, 1969), the findings for both Natives and Non-natives alike point to the salience of psychosocial attachment and other indicators of family functioning in affecting early onset drinking and drug use, behaviors well-recognized to lead to potentially adverse mental and physical health consequences as well as to negative social outcomes. PMID- 10842505 TI - Is the relationship between fertility of parents and children really weak? AB - The relationship between fertility of parents and children has been designated as "weak" by most investigators. This paper reviews the evidence over the past century and argues that, even allowing for problems with available data sources, the relationship was probably close to zero for pre-transitional populations. However, over time, the relationship has tended to become more substantial and is now of a similar order of magnitude in developed countries as other widely used explanatory variables. Possible mechanisms for the observed relationship are discussed, especially the roles of socialization and inherited factors. The types of data used are compared to the scientific questions posed, and the limitations of the common comparison of married-mother/married-daughter pairs are considered. Finally, some evidence from recent large-scale surveys in Britain and the United States is presented to show changes over recent periods and the relative effects of sibship size of fathers and mothers. PMID- 10842506 TI - New evidence for dysgenic fertility for intelligence in the United States. AB - Data were taken from the 1994 National Opinion Research Center survey of a representative sample of American adults to examine the relation between the intelligence of adults aged 40+ and their numbers of children and their numbers of siblings. The correlations were found to be significantly negative at -0.05 and -0.09, respectively, indicating the presence of dysgenic fertility. Further analysis showed that dysgenic fertility is present only in females. The correlation for females between intelligence and ideal numbers of children was effectively zero, indicating that if women had the numbers of children they consider ideal, dysgenic fertility would be reduced. PMID- 10842508 TI - Honey and sugar as a dressing for wounds and ulcers. PMID- 10842507 TI - On abandoning life when it is least difficult. AB - Suicide data from Sweden (1911-1993) and New Zealand (1975-1995) were used to test a hypothesis derived as an extension of the Seasonal Affective Disorder concept. Contrary to the hypothesis, but similar to what Durkheim had reported regarding Central Europe in the nineteenth century, suicides peaked in late spring (May in Sweden, November in New Zealand). Durkheim's hypothesis that there would be more monthly variation in suicides in rural areas was strongly supported. Our hypothesis is that people abandon life when it is least difficult because of a disjuncture between experience and expectation. PMID- 10842509 TI - Practical hospital pharmacy management: management strategies and methods. AB - This review deals with the practical management of hospital pharmacies based on experience in Indian Christian hospitals with capacities ranging from about 30 to 500 beds. However the standards of pharmacy management should not depend on size; they should be kept under review and improved as staff and financial considerations allow. PMID- 10842510 TI - Knowing when but not how!--mothers' perceptions and use of antibiotics in a rural area of Viet Nam. AB - Given the world-wide focus on how to rationally use antibiotics, national drug policy programmes have been developed in many countries in order to minimize the environmental antibiotic pressure and thereby hopefully limit increasing bacterial resistance. This study investigated perceptions of antibiotics in a health system with weak drug regulation. The study was conducted in two rural communes in Viet Nam, with a drug market characterized by the increased accessibility and consumption of pharmaceuticals. The study focused on rural mothers' perceptions and use of antibiotics in the treatment of acute respiratory infections (ARI) in children 5 years and under. A combination of qualitative and quantitative methods were used including key informant interviews, focus group discussions, and interviews with mothers and drug vendors. The study demonstrated that using a combination of qualitative and quantitative methods gives a better understanding of the prevailing perceptions and use of antibiotics in communities. The results showed that the mothers recognized well the signs of severe ARI and that antibiotics were reserved for more severe illness episodes, where penicillin V and ampicillin were first drugs of choice. However, the mothers' perceptions and use of antibiotics reflects indigenization of antibiotics into traditional Vietnamese thinking and medical practice. This resulted in self-medication and a respect for antibiotics from the mothers' point of view. A first step towards the rational use of antibiotics is already taken where mothers, as the health decision-maker, know when to initiate antibiotic treatment and try to limit unnecessary use of antibiotics. The next step is to develop a well-functioning health education programme in order to promote the correct use of antibiotics for a successful clinical outcome. This requires acknowledgement of the mothers' culture based behaviour. PMID- 10842511 TI - Laboratory quality control league. PMID- 10842512 TI - Hereditary congenital cataracts associated with sickle cell anaemia in a Nigerian family. AB - Pedigrees of hereditary disorders are rarely reported from sub-Saharan countries because of difficulties inherent in obtaining reliable family histories, case tracing and follow-up. This report relates to a four-generation family in northern Nigeria with dominantly inherited cataracts associated with recessively inherited sickle cell anaemia. It highlights the need for genetic counselling services in clinical practice in the region. PMID- 10842513 TI - Developments in the production and application of rabies vaccine for human use in China. PMID- 10842514 TI - Bone age determination in a paediatric population as an indicator of nutritional status. AB - The United Mission to Nepal Tansen Project is in the hill country, 300 km West of Kathmandu. This 40-year-old service centre's major emphasis has been on clinical and community medicine and education. Agronomy, forestry, water resources development, adult literacy, etc., have also been part of the project's outreach. We had long noted that the children of all of the various tribal groups in the area are small for their age as compared with children in developed countries. Fracture care X-rays were examined to reveal delay in ossification and in physeal closure. Using the standards found in the Radiographic Atlas of Skeletal Development of the Hand and Wrist, we quantified this apparent delay by determining the bone age of all patients presenting at the hospital with upper limb injuries requiring X-ray. The collection of data was limited to 100 days and to those under the age of 21. Two hundred and nineteen patients' data were analysed. We found that bone maturation delay was very common and often severe. The average bone age among the 219 was 28% below chronological age. That is, 60% of all fell two standard deviations below the norm: i.e. two-thirds of all 9 year olds had the expected skeletal development of 6 year olds in Boston, USA. Dental age was also analysed using deciduous tooth shedding and tooth eruption patterns. Dental age retardation averaged 21%, but a larger per cent had normal eruption dates. Other physical and sociological data were included in a questionnaire completed for each patient. Among the variables examined, poverty alone appeared to be the major mover. We believe that national small stature is a result of lifelong community dietary inadequacy. PMID- 10842515 TI - Tuberculous cervical lymphadenopathy: should antituberculous therapy be preceded by histological proof? AB - In this retrospective study, 212 patients with cervical lymphadenopathy (CL) were studied over a 4-year period (1987-1990). Ninety-six (45.3%) were found to be tuberculous, 64 (30.2%) had reactive hyperplasia, 31 (14.6%) had a malignant lymphoma, 12 (5.7%) showed metastatic carcinoma and nine (4.2%) were due to miscellaneous causes. The original provisional diagnosis was incorrect in 20 cases who were later found to have tuberculous lymphadenopathy (TL) and in 12 patients with secondary malignancy who were originally diagnosed as tuberculous. The study shows that treatment should be preceded by histological proof to avoid delay in treatment of serious malignant disorders, to avoid unnecessary antituberculous treatment in patients with benign reactive lymphadenopathy and to prevent delay of antituberculous treatment in tuberculous patients with atypical clinical presentations. PMID- 10842516 TI - Surgery and community participation in a community-based programme in war situations. AB - The paper presents the results and the experience gained from three 'mass surgical interventions' conducted in the Alpha Civil Hospital, Billing in south Sudan in the period between October 1996 and November 1997. The results showed that surgical services can be provided safely and economically even under most difficult conditions provided professional experience plus community commitment and participation are there. Such interventions lead to the development of strong community self-reliance and willingness which are fundamental if the programme is to be sustained. PMID- 10842517 TI - Envenoming by scorpions and snakes (Elapidae), their neurotoxins and therapeutics. PMID- 10842518 TI - Demographic characteristics and risk factors of first febrile seizures: a Jordanian experience. AB - A prospective study of 203 children admitted with a first febrile seizure was carried out over 18 months. Aseptic meningitis was diagnosed in nine children (4%). The mean age of the children was 19.9 months and the peak age incidence (88%) was between 6 months and 3 years. Complex seizures were seen in 16 cases (8%). There was a history of perinatal asphyxia in 16 patients (8%), positive family history of epilepsy in 3%, of first degree relatives and a history of febrile seizures in siblings in 13%. Upper respiratory tract infection was the commonest triggering factor, diagnosed in 53% of cases. The third child was the most commonly affected (22%) in the family. There was a lower incidence of both complex febrile seizures and history of seizures in siblings compared to other studies. Lumbar puncture should be considered for all children below the age of 2 years, presenting with a first febrile seizure. PMID- 10842519 TI - Build your own multipurpose, economically priced portable stereo microscope. AB - Binocular loupes and stereo microscopes are expensive items of equipment. However, inexpensive operating microscopes (ophthalmic) with coaxial illumination have been constructed for use in developing countries, where there is little medical funding. We describe one method of transforming economically priced pocket binoculars into a portable stereo microscope with nearly coaxial illumination. PMID- 10842520 TI - Audit of antimalarial chemotherapy in adult inpatients in Blantyre, Malawi. PMID- 10842521 TI - Cavernosal spongiosium shunt in the management of priapism in Zaria, Nigeria. AB - Between 1982 and 1994 13 patients have needed surgery for priapism persisting after conservative measures using the technique described by Quackel. PMID- 10842522 TI - Parotid tuberculosis--a clinical rarity. PMID- 10842523 TI - Claw hand as a complication of herpes zoster. PMID- 10842524 TI - Primary ovarian pregnancy: a rare and often missed diagnosis. PMID- 10842525 TI - Surgery in high-HIV incidence poor countries. PMID- 10842526 TI - Abdominal closure. AB - There have been considerable developments in abdominal closure over the past 30 years or so, and (happily for some of the poorer countries of the world) they have not largely involved undue expense. Indeed many of the developments are compatible with considerable cost-saving. I have changed over from the traditional (as it was then) method of multilayer closure with catgut to a one layer method with monofilament nylon (often fishing nylon) in 1971. Since that time, despite a previous unhappy experience with the upper abdominal paramedian closure, I have not had a single 'burst abdomen' in any adult patients since making that change. PMID- 10842527 TI - Managing sexually transmitted diseases in the tropics: is a laboratory really needed? PMID- 10842528 TI - Nairobi bomb blast--trauma and recovery. PMID- 10842529 TI - Prevalence of HIV, sexually transmitted disease and tuberculosis amongst new prisoners in a district prison, Malawi. PMID- 10842530 TI - Prevalence of intestinal parasites in rural areas in south Gugerat (India) PMID- 10842531 TI - Epidemiology of delayed initial contact between mother and newborn baby. PMID- 10842532 TI - Deaths from burns a major cause of surgical paediatric deaths in Zambia. PMID- 10842533 TI - Is screening for Toxoplasmosis appropriate in West Africa? PMID- 10842534 TI - Pelvic abscess in women in the tropics: a 10-year review in Enugu, Nigeria. PMID- 10842535 TI - Circumcision practice in Anjouan. PMID- 10842536 TI - Identifying malaria outbreak in clinical practice. PMID- 10842537 TI - Cholera-like diarrhoeal disease and rivers in rural Mozambique. PMID- 10842538 TI - Eusol management of burns. PMID- 10842539 TI - The need for the introduction of a course on drug management in pharmacy curriculum. PMID- 10842540 TI - Burns and skin grafting. PMID- 10842541 TI - Vitamin B6 supplementation with isoniazid therapy. PMID- 10842542 TI - The use of high-dose isomazid in intermittent regime TB treatment--some preliminary findings. PMID- 10842543 TI - Outbreak of Stevens-Johnson syndrome among Filipino overseas contract workers using mebendazole for helminthiasis prophylaxis. PMID- 10842544 TI - The ethics of clinical research in developing countries--a discussion paper. PMID- 10842545 TI - Practical hospital pharmacy management: drug distribution. PMID- 10842546 TI - Combined fluphenazine and lidocaine for pain relief in head and neck cancers. AB - A successful method of treating intractable neoplastic pain in 12 cases is discussed. A completely new method was tried combining long-acting phenothiazine and lidocaine to achieve long-lasting pain relief. This combination gave pain relief which lasted for weeks. A possible explanation of the mechanism is offered. PMID- 10842547 TI - Analysis of a simplified concentration sputum smear technique for pulmonary tuberculosis diagnosis in rural hospitals. AB - Rural hospitals in Africa face the challenge of an increasing number of pulmonary tuberculosis (PTB) cases, much of it related with the AIDS pandemic. The main diagnostic tool in these hospitals is the direct sputum smears. Sensitivity of direct sputum smears is low, especially in cases of AIDS related PTB. Several concentration methods have been described but none is routinely used in Zimbabwe. Fluorescence microscopy of auramine stained samples increases sensitivity and saves laboratory time but it is not an appropriate technique for rural African hospitals. In a rural hospital of southwest Zimbabwe, we studied the sensitivity and appropriateness of a simplified concentration method. The sensitivity against culture was significantly higher with the concentration method (80%) than with the classical direct smear method (57%). However, when sputum is not adequate (with high number of squamous epithelial cells, meaning upper airway origin), the sensitivity of this method is low. A combination of both methods is proposed to increase the sensitivity of PTB diagnosis in rural hospitals. PMID- 10842548 TI - Maternal mortality estimated using the Sisterhood method in Gulu district, Uganda. AB - A community-based retrospective maternal mortality study using the Sisterhood method was conducted in Gulu district between February and March 1996. The objectives were to estimate the magnitude of and identify factors associated with maternal mortality in the district. A total of 5522 adult respondents, randomly selected from 27 parishes, of the five counties in the district were interviewed. Between 1960-1996 324 maternal deaths occurred in the sisterhood sample. The maternal mortality rate (MMR) was estimated to be 662 per 100,000 deliveries [95% confidence interval (CI) 421-839 per 100 deliveries]. The leading causes of maternal death were: haemorrhage 45.1%; obstructed labour 26.2%; puerperal sepsis 9.6%; anaemia 2.2%; AIDS 2.2%; and gunshot wounds (GSW) 1.0%. Factors associated with maternal mortality included: age--31.8% of the mothers who died were below 20 years; education--57.1% had no formal education; 65% of the mothers had delivered at home, 50.6% had been attended to by untrained traditional birth attendants (TBAs), while 37.8% were attended to by relatives. The MMR was found to be 1.3 times higher than the estimated national MMR of 500 per 100,000 deliveries. Most maternal deaths (80.9%) were due to preventable causes, being related to low socioeconomic status and low-level education of women in the district. The intractable civil war in the district was a major underlying and contributory factor to the high maternal mortality in the area. A multifaceted approach to reduce maternal mortality in the district should target improving the socioeconomic conditions in the district with special emphasis on encouraging and supporting female education. Intensive education on maternal healthcare in antenatal clinics be conducted targeting husbands/spouses and relatives who care for the prenatal/pregnant and postnatal mothers. There is need for more trained TBAs per village who should be given effective support supervision. Ambulance transport services, motor and bicycle be made available at the district and community levels. At a national level the security situation should be improved in the district. PMID- 10842549 TI - Can plasma fructosamine substitute for glycated haemoglobin (HbA1c) estimation in the assessment of diabetic control? AB - The 'gold standard' marker of overall glycaemic control in diabetes mellitus is the level of glycated haemoglobin (HbA1c). It is, however, an expensive and technically difficult assay and is rarely appropriate to tropical laboratories. Plasma fructosamine measurement is cheaper and easier, though it reflects shorter term glycaemia. We have measured both indices of control in a group of 154 diabetic patients. There was close correlation between the two measurements (r = 0.6506, P < 0.001), but many patients with abnormal HbA1c levels had normal fructosamine levels. This resulted in an assay sensitivity (compared with HbA1c as gold standard) of only 30%, though specificity was 98%. We conclude that fructosamine measurement cannot be regarded as a substitute for HbA1c determination. PMID- 10842550 TI - Evaluation of formalin preserved allogeneic bone graft as a grafting material. AB - Formalin preserved allogeneic (FPA) bone graft was used in 18 patients with benign bone cavities, nine patients with malignant bone cavities, 12 patients with atrophic nonunion of fractures, seven with gap nonunion and 18 with comminuted fractures. The benign bone cavities were obliterated in 20 weeks and malignant cavities in 24 weeks. Union was obtained in atrophic ununited fractures in 29 weeks, in gap nonunion in 40 weeks and in comminuted fractures in 27 weeks. FPA is a simple, inexpensive and easily available alternative to autologous bone and to deep frozen bone in filling bone defects and enhancing fracture union. PMID- 10842551 TI - Drawover anaesthesia for cleft palate and lip surgery in Pokhara, Nepal. AB - We describe our experience and problems with 335 children who had general anaesthesia for cleft palate and lip surgery. Drawover anaesthesia using halothane and trichloroethylene was easy to use and safe. The main early postoperative problems were airway obstruction and bleeding. Later postoperative care in the ward was hampered by the lack of nursing care and monitoring. PMID- 10842552 TI - Neisseria gonorrhoeae infection among pregnant women in Peshawar, Pakistan: prevalence and risk factors. PMID- 10842554 TI - Maternal mortality: the figures--value and validity. PMID- 10842553 TI - The effectiveness of weekly iron supplementation in pregnant women of rural northern Malawi. AB - A weekly iron/folate supplement was compared with a standard daily iron/folate supplement in pregnant women living in rural Malawi. Women were enrolled as they attended the local antenatal clinic, stratified by grade of anaemia and then randomized to receive either 60 mg iron/0.25 mg folate per day (n = 211) or 120 mg iron/0.50 mg folate once a week (n = 202). Supplementation was continued for a minimum of 8 weeks (10 weeks on average) and was self administered by the women at home. Initial haemoglobin values for the daily (mu = 105.7 g/l) and weekly (mu = 104.4 g/l) groups as well as final haemoglobin values (107.5 g/l and 105.6 g/l, respectively) did not differ significantly between the two groups. Haemoglobin values increased by similar levels in both groups with the subset of anaemic women increasing by an average of 6.3 g/l in the daily group (n = 70) and 5.9 g/l in the weekly group (n = 66) for all women. For compliant, anaemic women, the increases were 7.4 g/l and 6.6 g/l for the daily and weekly groups, respectively. Compliance, as indicated by self reporting and by regular counts of remaining tablets, was significantly higher in the weekly group (76% compared with 60%, P < 0.05), however compliance was identical in both groups when assessed by a stool test for elemental iron. Reported side effects were significantly reduced in the weekly group (6% compared with 17%, P < 0.05). We conclude that a weekly iron supplement given to pregnant women in rural Malawi has similar haematologic effects, and an improved side effect profile, in comparison with a standard daily supplement when administered through an existing primary healthcare programme, although both regimens are relatively unsuccessful in the reduction of anaemia prevalence during pregnancy. PMID- 10842555 TI - Community financing of local ivermectin distribution in Nigeria: potential payment and cost-recovery outlook. AB - The preferred payment mechanism in a community financing scheme for local ivermectin distribution was elicited from randomly selected household heads from three communities in Nigeria using interviewer-administered structured questionnaires. The majority of the respondents in the three communities were prepared to pay for local ivermectin distribution. Additionally, the average amounts the respondents were prepared to pay per person treated ($0.28, $0.30 and $0.38 in Nike, Achi and Toro, respectively) were all more than the $0.20 ceiling recommended by the partners of the African Programme on Onchocerciasis Control (APOC). Thus, the cost-recovery outlook is bright in these communities. However, the preferred payment modality varied. Fee-for-service was the predominant payment modality in the Achi and Nike communities, while the Toro community preferred pre-payment. This study demonstrates that many communities have different payment preferences for endemic disease control efforts. This knowledge will help in developing acceptable and sustainable schemes. The imposition of unacceptable payment mechanisms will lead to an unwillingness to pay. PMID- 10842556 TI - Improving healthcare by perinatal mortality audit and feedback. PMID- 10842557 TI - A survey to gather sexually transmitted disease epidemiological and management data in the Solomon Islands. AB - A survey of 15 clinics and hospitals in the Solomon Islands (a South Pacific nation with a 45% rate of penicillin-resistant gonorrhoea) was undertaken to audit the quality of sexually transmitted diseases data collection, adherence to public health fundamentals and knowledge of the national gonorrhoea management guidelines. With the exception of one town clinic, data collection was limited, syphilis serological testing was low (28%) and correct knowledge of the national gonorrhoea treatment guidelines was also limited to 4/8 clinics (50%). Contact tracing was definitely undertaken in 2/8 (25%) of clinics. A high male/female notification ratio (3.6:1) for gonorrhoea was detected. Solomon Island clinics may be typical of other South Pacific Island communities in their inadequate attention to public health fundamentals. Every effort should be made to identify these shortcomings and provide feedback to clinic health workers to improve this service, particularly in the context of the more recent arrival of the HIV epidemic in the region. PMID- 10842558 TI - Hepatitis A seroprevalence rate in Syria. AB - Hepatitis A is a benign infection, which in the developing world affects mainly children; the majority of adults are immune by the age of 30. In the last decade or so, a shift in the prevalence pattern of hepatitis A virus (HAV) infection from a low- to a high-age group has appeared in the developed countries. This shift has been attributed to an improvement in the socio-economic and hygienic conditions. In the present study, 849 Syrians of all age groups and both sexes were tested by enzyme-linked immunosorbent assay technique for the seroprevalence of hepatitis A IgG antibody (anti-HAV IgG). It was observed that anti-HAV IgG was present in 89% Syrian population; with 50% in the 1-5 year age group and 95% in the 11-15 year age group. These results demonstrate that HAV infection in Syria is mostly acquired during childhood. As yet there is no serological evidence of a shift in HAV infection from a younger to a higher age group. PMID- 10842559 TI - Saturated aqueous sodium chloride solution for the removal of leeches. PMID- 10842560 TI - Appendicitis versus genital disease in young women in tropical Africa. AB - The differential diagnosis in women between acute appendicitis and acute gynaecological conditions on clinical examination is often difficult. A plea is made for the more widespread use of ultrasound and laparoscopy in the differential diagnosis of these conditions. PMID- 10842561 TI - Morbidity and mortality in prisons in Malawi. PMID- 10842562 TI - Complementary treatment of drug addicts in Malaysia. PMID- 10842563 TI - The effect of Qat chewing on blood pressure and heart rate in healthy volunteers. AB - The leaves of the Qat plant (Catha edulis Forsk., Celastraceae) which contain amphetamine like compounds are widely chewed in Yemen and East Africa for their pleasurable stimulant properties. There are also a number of unwanted side effects and this paper studies the effect on heart rate and blood pressure in 80 healthy volunteers. During a 3-h period of chewing fresh Qat leaves there was a significant and progressive rise in systolic and diastolic blood pressure and heart rate, and levels had not returned to baseline 1 h after chewing had ceased. Further studies are needed on possible cardiovascular morbidity associated with regular Qat chewing. PMID- 10842564 TI - Short bowel syndrome. PMID- 10842565 TI - Affection of cervical lymph nodes in rhinoscleroma. PMID- 10842566 TI - Pneumothorax complicating acute miliary tuberculosis in children. PMID- 10842567 TI - Heavy tapeworm infestation causing occlusive bowel syndrome in a severely psychiatrically disturbed man. PMID- 10842568 TI - Antimicrobial resistance in the tropics. AB - Bacterial resistance to antimicrobial agents is an increasing problem in many areas of the tropics. In most countries there is little information available to determine the patterns of resistance in different pathogens, nor are local data available to influence prescribing. This paper will review the development of antimicrobial resistance in the tropics, consider the current priority problems, and suggest strategies that may be taken to improve the surveillance of resistance. PMID- 10842569 TI - Methods of fluid administration for resuscitation and hydration under difficult circumstances: Part 1--The intravenous route. PMID- 10842570 TI - Head injury management (with no CT scanner). PMID- 10842571 TI - Voluntary screening for HIV. PMID- 10842572 TI - Management of burns. PMID- 10842573 TI - Chloroquine resistant malaria. PMID- 10842574 TI - Breastfeeding and asthma in children: findings from a West Australian study. AB - The primary aim was to determine whether there was an inverse association between the duration of exclusive breastfeeding and the development of traits associated with asthma in children at age six years. A prospective cohort study of children from Western Australia was enrolled prior to birth and followed to age six. Two thousand, nine hundred and seventy-nine children were recruited through antenatal clinics at the major tertiary obstetric hospital in Perth. Unconditional logistic regression was used to model the association between duration of exclusive breastfeeding and outcomes related to asthma or atopy at age six allowing for a number of important confounders. These included gender, gestational age, smoking in pregnancy and early child care. After adjustment for confounders, the introduction of milk other than breastmilk before four months of age was a significant (p < 0.05) risk factor for all asthma-related outcomes in six-year old children: (i) doctor diagnosed asthma odds ratio ?OR? = 1.25 (95% CI 1.02 1.54); (ii) wheeze three or more times since the age of one year OR 1.42 (1.15 1.76); (iii) wheeze in the last twelve months OR 1.28 (1.02-1.76); (iv) sleep disturbance due to wheeze within the last twelve months OR 1.41 (1.04-1.90); (v) age at doctor diagnosis (hazard ratio ?HR? 1.22 ?1.03-1.43?); (vi) age at first wheeze (HR 1.36 ?1.17-1.59?) and; (vii) positive reaction to common aeroallergens OR 1.27 (1.01-1.59). There is a substantial reduction in risk of childhood asthma as assessed at age six years, if exclusive breastfeeding is continued for at least the first four months of life. These findings are important for our understanding of the aetiology of and for the potential prevention of asthma in children. PMID- 10842575 TI - Cultural beliefs and breastfeeding duration of Thai working women. AB - Breastfeeding's crucial role in infant health is universally recognised and yet there are a number of influences on the duration of breastfeeding. This quantitative study undertaken in Thailand explored whether personal beliefs and values influenced Thai working women's duration of breastfeeding. Ninety-nine Thai working mothers were asked to complete a questionnaire to explore the relationship between their cultural beliefs and their breastfeeding practices. Although the majority of the women undertook techniques based on their cultural beliefs, which they believed would encourage breastfeeding practice, the findings demonstrate that the women's cultural beliefs had no significant effect on the duration of their breastfeeding. PMID- 10842576 TI - The implementation of the HOT program at the Royal Women's Hospital. AB - At the Royal Women's Hospital from November 1995, mothers and their infants were discharged on day three (a three-night stay) following a normal vaginal birth, if they were considered to be well and coping with early postnatal life. However, by March 1996, there were indications that not all mothers were managing breastfeeding as well as they needed to after discharge home. In an effort to reduce early avoidable breastfeeding problems, an extensive re-education program was undertaken. The 'Hands-Off Technique' Program (HOT) aimed to address the problem by educating the midwifery staff on ways to teach mothers about correct positioning and self-attachment of their babies without actually doing it for them. PMID- 10842577 TI - Ready for birth. PMID- 10842578 TI - An analysis of personal and social factors influencing initiation and duration of breastfeeding in a large Queensland maternity hospital. AB - This study aimed to determine the degree to which certain personal and social maternal factors, measured in the immediate postpartum period and during the next six months, were associated with the length of the breastfeeding experience. Qualitative and quantitative data were obtained from three questionnaires administered to 159 mothers, who delivered their infants within a three-month birth cohort at Royal Women's Hospital Brisbane during 1997. Interviews took place prior to hospital discharge, at three months postpartum and at six months postpartum. The study found that, while 91.1% of new mothers had breastfed their infants at least once, only 49.6% were breastfeeding at all by the time their infants were six months of age. Longer breastfeeding duration was most significantly associated with increased breastfeeding self-confidence, lower levels of anxiety and depression, increased self-esteem and coping capacity, and stronger social health. These findings have relevance to the content and process of antenatal and postnatal education programs undertaken with pregnant and postpartum women in all health care settings. PMID- 10842579 TI - Global problem of drug-induced hearing loss. AB - Clinically used drugs and chemical agents may potentially cause adverse effects to the human auditory and vestibular systems. Many of them, such as aminoglycosides and cisplatin, can play a critical role in the treatment of serious or life-threatening diseases; others, like loop diuretics or salycilates, offer such important therapeutical effects compared to the ototoxic side effects that the ototoxicity risk can be considered to be of minor importance. The problem of ototoxic side effects is more acute in developing countries, where highly effective and low-cost drugs are more easily prescribed without adequate monitoring. Medical awareness of doses, forms of administration, populations at risk, and possible synergism is necessary in order to develop appropriate care in the prescription of drugs with ototoxic side effects. Relatively recent issues such as risk-benefit analysis, patient-informed consent, and quality-of-life considerations, particularly when life expectancy can be low, are also to be considered. At present, a uniform method of monitoring for all potentially ototoxic therapeutics does not seem reasonable or practical. It is recommended, however, that individual auditory function be noted for a particular drug being employed. Protocols and exams should be easy, quick, sensitive, reliable, and as objective as possible. Benefits of audiological monitoring include the opportunity to change the patient's treatment course, improvement of patient and family awareness of the impact of hearing impairment, and timely prescription of amplification devices. Finally, particular attention should be paid to high-risk populations such as neonatal intensive care unit patients. PMID- 10842580 TI - Insights into ototoxicity. Analogies to nephrotoxicity. AB - The most common ototoxic compounds in clinical practice are aminoglycoside antibiotics, cisplatin, and loop diuretics (ethacrynic acid and furosemide). These agents also have substantial renal effects in the form of nephrotoxicity or diuresis. The understanding of these renal effects may provide insight into ototoxic mechanisms. For aminoglycosides, the renal proximal tubule cell is susceptible due to high concentration achieved and slow clearance with direct effects on phosphoinositide binding and mitochondrial bioenergetics. Pathogenesis appears to involve iron-induced free-radical formation, since iron chelators prevent nephrotoxicity. Analogous effects of aminoglycosides on the inner and outer hair cells have been observed. Cisplatin is also highly concentrated in the proximal tubule cell. Less is known about the direct toxic effects of this agent on renal cells. Insights into mechanisms or renal tubule cells could be directly relevant to the inner ear. The loop diuretics are direct inhibitors of the Na(+) K(+)-2Cl- cotransport system, which also exists in the marginal and dark cells of the stria vascularis, which are responsible for endolymph secretion. The ototoxicity of these agents may be indirect, due to changes in ionic composition and fluid volume within the endolymph. PMID- 10842581 TI - Free radicals and hearing. Cause, consequence, and criteria. AB - Reactive oxygen and nitrogen species, including free radicals, are produced in the human body in both health and disease. In health, they may arise as regulatory mechanisms, intercellular signaling species, or as bacteriocidal agents. Their production is normally controlled by the antioxidant defense mechanisms that include intracellular enzymes--for example, glutathione peroxidase and superoxide dismutase--and low molecular-mass compounds such as vitamin E or ascorbic acid. Although repair mechanisms exist, some steady-state basal oxidative damage occurs in all individuals. Oxidative stress arises when there is a marked imbalance between the production and removal of reactive oxygen and nitrogen species. This may originate from an overproduction of these substances or from a depletion in the antioxidant defenses. Certain drugs may induce oxidative stress by forming drug-derived radicals that can not only deplete the antioxidant defenses but can also react directly with biomolecules. To be able to assess whether oxidative stress is occurring in a particular tissue, reliable biomarkers of oxidative damage are required. Since oxidative stress can damage all major biomolecules in vitro and probably in vivo, biomarkers for DNA, protein, and lipid damage are being developed which, when taken with an assessment of the antioxidant status of the individual, will allow evaluation of the involvement of oxidative stress in the etiology of disease and in the side effects of drugs. There is some evidence to suggest that free radical mediated damage may be involved in the ototoxicity of aminoglycosides and cisplatin derivatives. Whether this is a cause or consequence of the toxic insult to the sensory hair cells of the inner ear remains to be determined. PMID- 10842583 TI - Calpain inhibitors as therapeutic agents in nerve and muscle degeneration. AB - It seems plausible to hypothesize that in all forms of neurodegeneration or other forms of tissue degeneration, a common pathway exists that, when deciphered, could lead to our understanding of a variety of diseases that result in tissue necrosis, as well as offer potential for therapeutic intervention. In recent years progress toward elucidating this common pathway has been accelerated through the studies of a number of laboratories, including our own, on the role of the protease calpain in this process. Thus, in a variety of disorders, such as stroke, spinal cord injury, traumatic nerve injury, Parkinson's disease, amyotrophic lateral sclerosis (ALS), Alzheimer's disease, muscular dystrophy, cataract formation, unregulated calpain proteolysis, initiated via dysregulation of calcium ion homeostasis, participates in the pathogenesis and is a potentially unifying mechanistic event. In order to demonstrate the feasibility of the approach we have taken in using the calpain inhibitor leupeptin as a therapeutic agent, I will describe two areas of research in which we have been engaged over the past 20 years. One is our long-standing interest in muscular dystrophy. The other is of more recent vintage, and involves the use of calpain inhibitors to protect sensory hair cells and spiral ganglion neurons from damage associated with acoustic trauma, this latter in collaboration with Dr. R. Salvi at SUNY Buffalo and Dr. A. Shulman at SUNY-Downstate. PMID- 10842582 TI - Molecular mechanisms of cell death in the ear. AB - Damage to sensory cells and neurones of the inner ear, induced by ototoxic drugs, physical stimulation, or even normal aging, occurs by mechanisms that are not yet fully elucidated. However, for the individual the consequent loss of hearing can be devastating. This the concept that, in some instances, this brain cell death may be preventable has provided an exciting and novel route to thwart the degenerative process. Signals that trigger cell death can be as diverse as the removal of essential growth factors, damage by exogenous toxins (including ototoxic drugs), free radicals, and excitotoxins. An important facet to the newly discovered death cascade is that it can be halted, and such interventions may rescue the dying cell. The question now remains whether any of the cell death observed on ototoxic or noise-induced hearing loss occurs by an "active" programmed mechanism (apoptosis), as physiological cell death does in the developing ear, and if so, whether it can be prevented. PMID- 10842584 TI - Acute and chronic effects of aminoglycosides on cochlear hair cells. AB - The first detectable effect on the auditory system after a single high-dose injection of an aminoglycosidic antibiotic (AA) like gentamicin (GM) is the reversible blockade of medial efferent function, probably via blockade of calcium channels at the base of the outer hair cells (OHC). The kinetics of this effect are compatible with that of the molecule in perilymph. In the course of chronic treatment with lower doses, however, ototoxicity develops only after several days of treatment. Still GM can be observed inside the OHCs as soon as 24 hours after the first injection, and will be still present in some OHCs as long as 11 months after a chronic, nonototoxic 6-day treatment. In vitro, the short-term viability of isolated OHCs is not affected by exposure to AAs, but their transduction channels and their response to acetylcholine are reversibly blocked. However, developing organs of Corti in culture are highly and rapidly affected by exposure to AAs. Yet during direct intracochlear perilymphatic perfusion of GM, 2-mM solutions are not ototoxic, and with perfusion with a 20-mM solution ototoxicity develops only after several days of perfusion. From these various observations one can describe some aspects of the mechanisms of ototoxicity of AAs, from their access to perilymph and endolymph, to penetration in the hair cells, likely via endocytosis at their apical pole, and intracellular cytotoxic events. PMID- 10842585 TI - Normal and altered cytoarchitecture of the inner ear. AB - Scanning electron microscope gave new insights into real morphology of the inner ear and has enhanced basic research. In fact, during the last twenty years, the cellular system that composes the organ of Corti and the vestibular apparatus has been clearly described by many authors. We now have a better understanding of each cell type and that they have a physiological functioning as ear and vestibular system. In addition, new insights into the damage caused by many drugs, such as aminoglucosides, can be investigated by studying the morphological changes and the reparative process following the injury. Animals such as guinea pigs can be used as ideal models to investigate the alterations in the sensory epithelium of the cochlear and vestibular receptors following ototoxic poisoning. PMID- 10842586 TI - An animal model based on the Sprague Dawley rat for the evaluation of ototoxicity. AB - Various methodological approaches that can be used to detect ototoxic effects caused by the administration of various substances are presented, using the Sprague-Dawley rat as an animal model. Electrophysiological data are also presented to show how the model behaves with potentially ototoxic (hyaluronic acid) and initially inert (hydroxy-propyl-methyl-cellulose) substances. PMID- 10842587 TI - Genetic factors in aminoglycoside toxicity. AB - Ototoxicity is the major irreversible toxicity of aminoglycosides, and occurs both in a dose-dependent and idiosyncratic fashion. The idiosyncratic pathway is presumably due to genetic predispositions, and in 1993 we identified an inherited mutation that predisposes to aminoglycoside ototoxicity, the A1555G mutation in the mitochondrial 12S ribosomal RNA gene. Seventeen-33% of patients with aminoglycoside ototoxicity carry this mutation. In a search for additional susceptibility mutations, a dual strategy of yeast genetics and candidate genes was employed. Through yeast genetics 8 genes that by overexpression prevent aminoglycoside toxicity were identified. The human homologues of these genes may harbor aminoglycoside susceptibility mutations. Another candidate gene is the mitochondrial ribosomal protein S12, which interacts with the ribosomal RNA gene and in bacteria can harbor aminoglycoside resistance mutations. Analysis of this gene in 41 patients with aminoglycoside ototoxicity did not reveal any mutations in the coding regions. However, an Italian family with five maternally related family members who went deaf after aminoglycosides led to the identification of the second susceptibility mutation in the mitochondrial 12S ribosomal RNA gene, delta T961Cn. While these findings have immediate clinical implications for prevention of aminoglycoside-induced ototoxicity, they have not yet led to a clear understanding of the pathophysiology of aminoglycoside toxicity or the development of therapeutic options after the onset of symptoms. PMID- 10842588 TI - A missense mutation in myosin VIIA prevents aminoglycoside accumulation in early postnatal cochlear hair cells. AB - Myosin VIIA is expressed by sensory hair cells in the inner ear and proximal tubule cells in the kidney, the two primary targets of aminoglycoside antibiotics. Using cochlear cultures prepared from early postnatal Myo7a6J mice with a missense mutation in the head region of the myosin VIIA molecule we show that this myosin is required for aminoglycoside accumulation in cochlear hair cells. Hair cells in homozygous mutant Myo7a6J cochlear cultures have disorganized hair bundles, but are otherwise morphologically normal and transduce. However, and in contrast to hair cells from heterozygous Myo7a6J cultures, the homozygous Myo7a6J hair cells do not accumulate [3H]gentamicin and do not exhibit an ototoxic response on exposure to aminoglycoside. Possible roles for myosin VIIA in the process of aminoglycoside accumulation are discussed. PMID- 10842589 TI - Antioxidant therapy attenuates aminoglycoside-induced hearing loss. AB - Aminoglycosides have been a mainstay in antibacterial therapy for over 50 years. Worldwide today, they are the most commonly used antibiotics because of high efficacy and low cost. However, nephrotoxicity and ototoxicity are two major side effects. Prevention of these side effects has been attempted for several decades, but no clinical treatment has yet been realized. A therapeutic approach based on a novel hypothesis of the toxic mechanism of aminoglycosides is proposed. Aminoglycosides chelate iron, and the resulting iron-aminoglycoside complex is redox-active, catalyzing the formation of free radicals. The prevention of ototoxicity with antioxidants and iron chelators has been successfully demonstrated in guinea pigs. Both hearing loss and vestibular dysfunction induced by several aminoglycosides were significantly attenuated without compromising the antibacterial efficacy of the drugs. This finding may lead to a safe and effective way to eliminate the threat of ototoxicity of this commonly used class of antibiotics. PMID- 10842590 TI - Neurotrophins, NMDA receptors, and nitric oxide in development and protection of the auditory system. AB - Neurotrophic factors are secreted peptides that when interacting with specific classes of membrane receptors activate intracellular signaling cascades that prevents neuronal death during embryonic development. The establishment of gene targeted null mutant mice for the neurotrophic factors, brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT3) has led to the discovery that they are crucial trophic factors for the survival of auditory and vestibular neurons during development. BDNF is the major survival factor for vestibular ganglion neurons, while NT-3 only support a small number of these neurons. In the cochlea, auditory type I neurons require NT3 for their survival, whereas type II neurons depend on BDNF. With this information at hand recent progress has been made regarding the prevention of aminoglycoside-induced hearing loss in the adult guinea pig. These results and the mechanisms leading to hair cell damage are discussed in this paper. PMID- 10842591 TI - Ototoxicity. Amelioration by protective agents. AB - The findings of studies from this laboratory are summarized to compare the efficacy of four chemoprotective agents against the effects of cisplatin-induced hearing loss and biochemical damage in the rat cochlea. A number of studies have shown that cisplatin is ototoxic, resulting in hearing loss, morphologic damage, and biochemical changes in the cochlea. These studies used Wistar rats, which underwent pre- and posttreatment ABR testing using clicks and tonebursts stimuli at 8, 16, and 32 kHz. Controls received i.p. saline injection. Cisplatin-treated rats were given 16 mg/kg cisplatin i.p. Animals received protective agents in the following dosage: DDTC protected rats received 600 mg/kg subcutaneously an hour after cisplatin. MTBA-protected animals were given 250 mg/kg i.p. 30 minutes before cisplatin. Animals protected with ebselen received 16 mg/kg i.p. an hour before cisplatin. One hundred mg/kg of alpha-lipoic acid was injected i.p. 30 minutes before cisplatin. Rats were sacrificed three days after treatment and the cochleae were harvested and frozen in liquid nitrogen and stored at -80 degrees C until analysis of glutathione (GSH), the activity of antioxidant enzymes (superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase) and malondialdehyde was performed. Cisplatin-treated rats were found to have ABR threshold shifts of 27-40 dB, and rats treated with chemoprotective agents plus cisplatin all had ABR thresholds shifts of less than 10 dB. Significant depletion of glutathione and decrease of the activities of the antioxidant enzymes were observed in cisplatin-treated rats. These changes were accompanied by a marked elevation of malondialdehyde. These changes were almost completely prevented by the use of the chemoprotective agents. These findings suggest that cisplatin ototoxicity is related to lipid peroxidation and that the use of protective agents prevents hearing loss and lipid peroxidation by sparing the antioxidant defense system in the cochlea. PMID- 10842592 TI - Selective loss of inner hair cells and type-I ganglion neurons in carboplatin treated chinchillas. Mechanisms of damage and protection. AB - Carboplatin preferentially destroys inner hair cells (IHCs) and type-I spiral ganglion neurons while sparing outer hair cells (OHCs). Loss of IHCs and type-I ganglion cells is associated with a significant reduction of the compound action potential (CAP). However, the cochlear microphonic (CM) potential and distortion product otoacoustic emissions (DPOAEs) remain normal, indicating that the OHCs are functionally intact. In the vestibular system, carboplatin selectively destroys type-I hair cells and their afferent neurons. Damage of type-I vestibular hair cells and their afferent terminals is associated with significant depression of nystagmus induced by cold, caloric stimulation. Histochemical studies revealed a rapid decrease in succinate dehydrogenase (SDH) staining in IHCs soon after carboplatin treatment, and staining intensity remained depressed in surviving IHCs for at least 1 month after carboplatin treatment. These results suggest that carboplatin depresses the metabolic function in surviving IHCs. Several lines of evidence suggest that free radicals may contribute to carboplatin-induced sensory cell damage. Intracochlear infusion of L-buthionine [S,R]-sulfoximine (BSO), which depletes intracellular glutathione (GSH), increases IHC and OHC loss. Previous in vitro studies have shown that neurotrophin 4/5 (NT-4/5) promotes the survival of spiral ganglion neurons from cisplatin ototoxicity. In vivo perfusion of NT-4/5 promoted the survival of spiral ganglion neurons, but did not protect the hair cells. PMID- 10842593 TI - A radical demise. Toxins and trauma share common pathways in hair cell death. AB - The pathologic similarities noted after ototoxic and/or traumatic injury to the cochlea as well as the key features of the cochlea that make it susceptible to reactive oxygen species (ROS) damage are reviewed. Recent evidence linking ROS to cochlear damage associated with both ototoxins and/or trauma are presented. Mechanisms of generation of ROS in the cochlea and how these metabolites damage the cochlea and impair function are also reviewed. Finally, examples of novel therapeutic strategies to prevent and reverse hearing loss due to noise and/or ototoxins are presented to illustrate the clinical relevance of these new findings. PMID- 10842594 TI - Protection and spontaneous recovery from cisplatin-induced hearing loss. AB - Cisplatin [cis-diamminechloroplatinum(II)] has proved itself as a potent antineoplastic agent. However, nephrotoxicity, neurotoxicity, gastrointestinal toxicity, myelosuppression, and ototoxicity interfere with its therapeutical efficacy. Forced diuresis reduces nephrotoxicity, effectively leaving neurotoxicity and ototoxicity as the major side effects of concern, and gastrointestinal toxicity and myelosuppression as the secondary side effects. So far, attempts to reduce these side effects by developing equally potent platinum analogs have been unsuccessful. Some success has been achieved, however, by co treatment with protective agents. Nearly all these agents are sulfur- or sulfhydryl-containing compounds (thio compounds), known as antioxidants and potent heavy metal chelators. These thio compounds may provide protection from cisplatin toxicity either (1) by direct interaction between the cisplatin and the thio moiety, (2) by displacing platinum from its site of toxic action, (3) by preventing platinum from interfering with superoxide dismutase, or (4) by scavenging of cisplatin-induced free radicals. In particular the first two protective mechanisms bear the risk of reducing the antineoplastic activity of cisplatin. Since nephrotoxicity can be controlled effectively by forced diuresis, a more specific approach of coping with ototoxicity by focusing on protection at the sensorineural level was chosen. Being familiar with the neuro-protective and neurotrophic properties of ACTH-related neuropeptides, specifically against cisplatin-induced peripheral neuropathies, it was judged expedient to test for a possible otoprotective action of these neuropeptides. The results were positive, although tainted with high interanimal variability. When testing for the possibility that the neuropeptides would merely delay cisplatin-induced ototoxicity rather than reduce it, it was discovered in control series without neuropeptide co-treatment that the ear can recover spontaneously from cisplatin induced hearing loss. This was found both electrophysiologically and in outer hair cell (OHC) counts. Although these preliminary findings require further investigation, they strongly suggest that spontaneous recovery of cochlear injury can occur in the mature mammalian cochlea. Moreover, the otoprotective action of the ACTH-related neuropeptides suggests that it may be possible to stimulate recovery from acute hearing loss using neuropeptides. PMID- 10842595 TI - Evaluation of cisplatin ototoxicity in a rat animal model. AB - The ototoxic effects of cisplatin were evaluated by otoacoustic emissions and evoked auditory responses. A transient otoacoustic emissions protocol indicated no significant ototoxic effects in rats treated intravenously with 7.5 mg/kg/week for 2-weeks. A chronic 6-week treatment (2.5 mg/kg/week) monitored by 2F1-F2 distortion product emissions presented significant SNR alterations in a narrow range of frequencies (5.04-5.66 kHz). An acute treatment of 15 mg/kg, using slow 30-min intraperitoneal infusion, presented the highest DP and ABR alterations. The SNR at the 2F1-F2 frequencies 6.34, 7.13, and 7.56 kHz was found significantly decreased, and ABR latency measurements from 8-kHz burst stimuli verified these alterations. PMID- 10842596 TI - Mechanisms of alterations in the microcirculation of the cochlea. AB - Labyrinthine function is tightly coupled to proper homeostasis. This includes appropriate blood flow that is under strict autoregulatory control. Perturbations in labyrinthine microcirculation can lead to significant cochlear and vestibular dysfunction. The etiology of many otologic disorders, including sudden sensorineural hearing loss, presbyacusis, noise-induced hearing loss, and certain vestibulopathies, are suspected of being related to alterations in blood flow. Some of the mechanisms responsible for hypoperfusion and possibly ischemia, within the cochlea, are addressed, with emphasis on the possibility that both noise and age contribute to localized low blood-flow states and stasis. This reduction in blood supply to the cochlea is likely, in part, responsible for reduced auditory sensitivity associated with chronic noise exposure and aging. PMID- 10842597 TI - Successful treatment of noise-induced cochlear ischemia, hypoxia, and hearing loss. AB - Cochlear blood flow (CoBF), perilymphatic partial pressure of oxygen (PL-pO2), cochlear microphonics (CMs), compound action potentials of the auditory nerve (CAPs), and auditory brainstem responses (ABRs) were studied in noise-exposed guinea pigs during and after the following treatments: intravenous infusion of isotonic saline (placebo); blood flow promoting drugs (hydroxyethyl starch = HES, pentoxifylline, betahistine, gingko biloba, naftidrofuryl); antiinflammatory agents (prednisolone, diclofenac sodium, histamine H1-receptor antagonist); isobaric oxygenation (IBO); and hyperbaric oxygenation (HBO) with and without supplements (simultaneous infusion of isotonic saline, pentoxifylline, prednisolone, or HES). It was found that PL-pO2 declined simultaneously with deterioration of CM, CAP, and ABR amplitudes after exposure to broad-band noise (bandwidth 1-12 kHz, 30 min, 106-dB SPL). CoBF decreased only 30 min after cessation of broad-band noise and progressed with cochlear hypoxia, while the hearing loss showed no further signs of deterioration and no recovery up to 3 h after exposure. Treatment (60 min) started 60 min after cessation of noise and was studied for a further 60 min. Isotonic saline did not influence the measuring parameters. Noise-induced cochlear hypoxia was compensated by IBO and more effectively by HBO with and without supplements, while other treatments had no sustained effect. A sustained therapeutic effect on noise-induced cochlear ischemia was achieved only by HES, HBO + HES, and pentoxifylline. However, the best therapeutic effect on noise-induced hearing loss was achieved with a combination of HBO and prednisolone, followed by monotherapy with prednisolone or HES with the result that not only did the CAPs and ABRs completely recover, the CMs also showed significant improvement, although full recovery did not occur. All other therapies were significantly less effective or did not improve noise induced reduction of auditory evoked potentials. PMID- 10842598 TI - Excitotoxicity, synaptic repair, and functional recovery in the mammalian cochlea: a review of recent findings. AB - Besides its fast excitatory properties, glutamate is known to have neurotoxic properties when released in large amounts or when incompletely recycled. This so called excitotoxicity is involved in a number of acute and/or degenerative forms of neuropathology such as epilepsy, Alzheimer's, Parkinson's, stroke, and retinal ischemia. In the cochlea, excitotoxicity may occur in two pathological conditions: anoxia and noise trauma. It is characterized by a two-step mechanism: (1) An acute swelling, which primarily depends on the AMPA/kainate type of receptors, together with a disruption of the postsynaptic structures (type I afferent dendrites) resulting in a loss of function. Within the next 5 days, synaptic repair may be observed with a full or a partial (acoustic trauma) recovery of cochlear potentials. (2) The second phase of excitotoxicity, which may develop after strong and/or repetitive injury, consists of a cascade of metabolic events triggered by the entry of Ca2+, which leads to neuronal death in the spiral ganglion. Ongoing experiments in animals, tracking the molecular basis of both these processes, presages the development of new pharmacological strategies to help neurites to regrow and reconnect properly to the IHCs, and to prevent or delay neuronal death in the spiral ganglion. Human applications should follow, and a local (transtympanic) strategy against cochlear excitotoxicity may, in the near future, prove to be helpful in ischemic- or noise-induced sudden deafness, as well as in the related tinnitus. PMID- 10842599 TI - Recovery of kainic acid excitotoxicity in chinchilla cochlea. AB - The present study examines the recovery of the inner hair cell (IHC)/auditory nerve synapse following cochlear excitotoxicity induced by kainic acid (KA). Three hours after KA treatment, there was massive swelling of type I afferent endings under the IHCs. Five to ten days later, the pattern of IHC innervation appeared to be normal. Distortion-product otoacoustic emissions were normal during the whole experiment. The amplitude of the auditory nerve compound action potential (CAP) was significantly reduced immediately after KA treatment and then recovered over a 30-day period. However, it only took five days for the evoked response from the inferior colliculus (IC) to recover from a substantial depression. In contrast to amplitudes, thresholds for the CAP and IC recovered at the same rate and returned to normal within 5 days after KA. Single auditory nerve fibers were also assessed at various times after the KA treatment. Ten days after KA, these fibers had almost normal thresholds, tuning, spontaneous, and driven discharge rates. The results indicate that (1) excitotoxically damaged cochlear afferent neurons can rapidly regenerate and establish viable synapses with the IHCs, and (2) the central auditory system recovers more rapidly than the periphery. PMID- 10842600 TI - Glial cell line-derived neurotrophic factor. Potential for otoprotection. AB - Sensorineural hearing loss results from the degeneration of hair cells and/or auditory neurons in the cochlea of the inner ear. BDNF and NT-3 were shown to support survival of auditory neurons both in vitro and in vivo. Cochlea from P3 P4 rats were cultured as floating explants and hair cells in the organ of Corti were identified by phalloidin-FITC immunostaining. Treatment with cisplatin (35 micrograms/mL) or neomycin (0.6 mM) resulted in 21.2 +/- 6.0% and 7.4 +/- 4.7% surviving hair cells, respectively, after 3 days in culture. GDNF, added together with the ototoxins, increased their number to 46.7% and 37.4%, respectively. In cultures of dissociated cochlea from 4-week-old rat, cisplatin (5 mg/mL) added 24 h after seeding resulted in only 6.1 +/- 1.2% surviving neurons. However, when cisplatin was added together with GDNF (10 ng/mL), 32.8 +/- 1.0% of the neurons survived. The efficacy of GDNF in animal models of ototoxicity was tested next. Guinea pigs were pretreated with GDNF in one ear, delivered either by infusion into the inner ear (scala tympani) with Alzet minipumps (50 ng/mL at a 0.5 microL/h), or injected into the middle ear (120 microL at 1 mg/mL) through the tympanic membrane. The ear that did not receive GDNF always served as control. Ototoxicity was induced systemically either by intraperitoneal cisplatin injections (1 mg/kg/day for 15 days or two injections of 7.5 mg/kg at a 5-day interval or by a combination of kanamycin (200-300 mg/kg, administered subcutaneously) and ethacrinic acid (40 mg/kg, intravenous). It was found that the number of surviving hair cells in GDNF-treated ears was about twice that of control ears in animals exposed to the ototoxins. The transducing GDNF receptor (ret) is expressed in the inner ear. PMID- 10842601 TI - Neurotrophic factors in the auditory periphery. AB - Many of the neurotrophic factors promote the survival of developing peripheral sensory neurons, and they might be useful as therapeutic agents in the adult neuronal systems. During development, neurotrophin-3 (NT-3) and brain-derived neurotrophic factor (BDNF) mRNAs are expressed in the auditory sensory epithelium, which composes the peripheral target field of the cochlear (spiral) neurons. NT-3 mRNA is also expressed in the inner hair cells of the mature organ of Corti. mRNAs encoding their signal-transducing receptors, TrkC and TrkB, respectively, are expressed in the cochlear neurons. In addition to neurotrophins, which seem to have an important role during development, another neurotrophic factor, glial cell-line-derived neurotrophic factor (GDNF), seems to be involved in the maintenance of postnatal auditory neurons. In the present work, the cellular distribution of neurotrophins and GDNF in the developing and adult rat inner ear are compared. The effects of recombinant neurotrophins and GDNF on dissociated cochlear neurons in vitro are also compared. Recently, NT-3 and BDNF were used in vivo as therapeutic agents to protect guinea pig cochlear neurons from aminoglycoside-induced degeneration and GDNF from noise-induced degeneration. These data demonstrate that NT-3, BDNF, and GDNF might be potential candidates for prevention of degeneration of the auditory nerve in man. PMID- 10842602 TI - Rescue and regrowth of sensory nerves following deafferentation by neurotrophic factors. AB - Trauma and loss of cochlear inner hair cells causes a series of events that result first in the retraction of the peripheral processes of the auditory nerve, scar formation in the organ of Corti, and over the course of weeks to months (depending on the species) the loss of auditory nerve cell bodies (spiral ganglion cells). Neurotrophic factors play an important role in the mature nervous system as survival factors for maintenance and protection and also can play a role in regrowth. Studies in the cochlea now show that application of exogenous neurotrophic factors can enhance survival of spiral ganglion cells after deafness and induce regrowth of peripheral processes, perhaps by replacing lost endogenous factors. Combinations of factors may be most effective for achieving greatest survival and regrowth. Our studies find that brain-derived neurotrophic factor (BDNF) and glial-line-derived neurotrophic factor (GDNF) are very effective at enhancing spiral ganglion cell survival following deafness from ototoxic drugs or noise. It has also been found that BDNF plus fibroblast growth factor (FGF) is very effective at inducing process regrowth. Electrical stimulation also acts to enhance spiral ganglion cell survival, and the combination of electrical stimulation and neurotrophic factors could prove a most effective intervention. PMID- 10842603 TI - Role of neurotrophins and lectins in prevention of ototoxicity. AB - Degeneration of hair cells (HC) and/or spiral ganglion neurons (SGN) is a major cause of hearing loss. Postnatal rat cochlear explant cultures are used to study the toxic actions of different classes of ototoxins and to identify molecules that can protect SGN and HC from ototoxic damage. Various ototoxins induce differential damage to HC and/or SGN. While gentamicin preferentially causes HC death, sodium salicylate selectively induces degeneration of SGN. In contrast, cisplatin results in destruction of both SGN and HC. Specific neurotrophins, including NT-4/5, BDNF, and NT-3, greatly protect SGN from all three types of ototoxins. In contrast, NGF and other growth factors have no effect. Of the 51 compounds examined, only concanavalin A (Con A), a lectin molecule, significantly protects HC from gentamicin. A dose-dependent study of Con A shows that maximal protection occurred at 100 nM. Further experiments indicates that preincubation of Con A with gentamicin does not form a complex, and coaddition of Con A and gentamicin to bacterial cultures, such as E. Coli cultures, does not interfere with the antibiotic activity of gentamicin. When the other 21 lectins are examined, Erythrina cristagalli lectin and Detura stramonium lectin also show activity similar to Con A. These findings may help elucidate the mechanisms of ototoxins and suggest that specific neurotrophins and lectins may be of therapeutic value in the prevention of ototoxin-induced hearing loss. PMID- 10842604 TI - Treatment of acoustic trauma. AB - The aim of this study is to determine the actual efficiency of the present medical treatments of acoustic trauma. Untreated and treated groups of guinea pigs are exposed to a traumatic noise exposure, inducing up to 60-dB threshold shift. The recovery is followed for up to 14 days. The first results indicate that in some animals the recovery of the threshold shifts are complete despite the fact that significant areas of hair cells are damaged. The most widely used medical treatments of acoustic trauma (oxygenotherapy, carbogen, hyperbaric oxygen, vasoactive agents, and corticotherapy) have been tested. Preliminary results indicate that pure oxygen and carbogen seem ineffective, hyperbaric oxygen used alone is dangerous, and corticoids and combined corticoid/hyperbaric oxygen seem to improve functional and morphological recovery. This study will be taken as a reference to look for new treatments that will be applied directly to the cochlea by means of an implanted osmotic micropump. PMID- 10842606 TI - Protective effect of the cochlear efferent system during noise exposure. AB - The aim of the present study was to confirm the hypothesis that the cochlear efferent system is involved in the mechanisms underlying the "toughening" effect at high frequencies. The toughening effect is defined as a progressive threshold shift reduction when repeated exposures to the same noise are applied. Vestibular neurectomy was performed through a posterior cranial fossa approach in six healthy pigmented guinea pigs, and it assured the interruption of both crossed and uncrossed olivocochlear bundles to one ear only, before their entrance in the internal auditory meatus. The animals were then implanted with permanent electrodes for the electrocochleographic findings. Ten days after the operation the animals were exposed to octave-band toughening noise, centered at 4 kHz, at 85-dB SPL, for 10 consecutive days, 6 hours on/18 hours off. The hearing threshold was registered before and at the end of each exposure session. The behavior of the hearing threshold in the operated ears was then compared to that of the controlateral, nonoperated ears. Complete recovery from TS in the control ear began after four days of exposure, whereas in the operated ear hearing loss increased to day 7 (55 dB), with only a partial reduction (45 dB) beyond ten days of exposure. The results of the present study clearly demonstrated that sectioning of the OCB in guinea pigs causes persistent hearing loss during noise exposure conditioning, in comparison to the contralateral, nonoperated ear. Thus, one can assume that the lack of decrease of TS during intermittent noise exposure could be due to the loss of the protective effect of the efferent fibers, perhaps mediated by the lateral OC neurons that synapse beneath the IHCs. PMID- 10842605 TI - Gene therapy in the inner ear. Mechanisms and clinical implications. AB - The application of gene therapy to the inner ear is an emerging field of study. Most studies report the expression of marker genes (e.g., galactosidase) within the tissues of the cochlea. The first biologic response of an inner ear tissue (i.e., auditory neurons) to transduction by a gene therapy vector expressing a therapeutic gene (a herpes amplicon vector containing a BDNF gene) was observed in spiral explants obtained from early postnatal rat cochleae. This study was important because it demonstrated the feasibility of a gene augmentation approach to treat traumatized cochleae. Long-term expression of transduced or transfected genes in cochlear tissues have been obtained with adenovirus, adeno-associated virus, and herpes amplicon vectors. The herpes amplicon vector (i.e., HSVbdnflac) that evoked a biologic response in vitro has also been successfully used to support the survival of auditory neurons in vivo following loss of the auditory hair cells (i.e., loss of trophic factor). Gene therapy has been successfully applied to the cochlea of a laboratory animal, and future studies will define the types of vectors and therapeutic genes that will work best for the treatment of inner ear diseases in the clinic. PMID- 10842607 TI - The role of antioxidants in protection from impulse noise. AB - The hearing loss from exposure to noise and ototoxic drugs share a number of audiological and pathological similarities. Recent research has shown that reactive oxygen species (ROS) may be a common factor in both noise- and drug induced hearing loss. This review describes three experiments that point to ROS as a causative factor in both noise- and drug-induced hearing loss and antioxidants as a protective agent. In the first experiment, the ears of chinchillas were treated with R-N6-phenylisopropyladenoisine (R-PIA) and exposed to 150-dB impulse noise. The treated ears developed substantially less permanent hearing loss (PTS) and hair cell loss than the untreated ears. One interpretation of this experiment is that R-PIA increases the availability of glutathione (GSH). In the second experiment, the role of GSH was specifically examined. The ears of chinchillas were treated with glutathione monoethylester (GEE), a pro-GSH drug that has been shown to readily cross cell membranes and increase GSH levels. The GEE-treated ears had significantly less PTS and hair-cell loss than the nontreated ear. Previous research has shown that moderate levels of noise exposure can increase a subject's resistance to noise, and also increase the availability of antioxidant enzymes in the cochlea. In the third experiment, chinchillas were given a series of "toughening" exposures (i.e., 6 h of a 0.5-kHz OB noise at 95 dB for 10 days). After the series of "toughening" exposures, the subjects were treated with carboplatin, a drug that causes massive inner-hair cell lesions in the chinchilla. The animals receiving the 10-day toughening exposure developed less PTS and hair-cell loss than the control animals. PMID- 10842608 TI - Gender-specific effects of drugs on hearing levels of older persons. AB - As part of a study of human presbyacusis, a questionnaire on medicinal drug usage was given to 357 subjects (184 females, 173 males). Previous results from 211 subjects showed gender effects, that is, for males, none of the drugs had any measurable effects on hearing, whereas women taking calcium channel blockers (CCBs) had hearing levels 12 dB better than women not taking them; women taking beta adrenergic medication had hearing levels 20 dB poorer, and women taking antihistamine/cold preparations had hearing levels 9 dB poorer. Results from the original 211 subjects were confirmed when the sample size was increased from 211 to 357 subjects only for the beta adrenergic medications. Results for antihistamine/cold preparation medications showed small effects only for female subjects. Data from 13 additional female subjects who used CCBs showed hearing levels 10-14 dB poorer than predicted from the original data. Male data were consistent in both samples. The inconsistency for females could reflect sampling error. A more likely possibility is that since the original 10 subjects using CCBs had a mean age of 72 yr and the second sample of 13 had a mean age of 79.5 yr, poorer hearing levels might be anticipated because of the difference in chronological age and possibly duration of drug usage. PMID- 10842609 TI - Dynamic studies of ototoxicity in mature avian auditory epithelium. AB - Hearing loss induced by ototoxicity is a worldwide problem despite the development of newer antibiotics and chemotherapy agents. The cellular mechanisms responsible for aminoglycoside-induced hearing loss are still poorly understood. We have developed two different methods of studying the dynamic cellular and subcellular changes in the chick auditory sensory epithelium that occur during hair cell death. The first study was performed in mature chicks after a single, high dose injection of gentamicin, which results in the rapid loss of all hair cells in the basal third of the cochlea. Chicks were sacrificed at discrete time points after drug treatment, and transmission electron microscopy was performed to study the ultrastructural changes in basal hair cells during the course of cell death. We noted various changes in the cell morphology including accumulation of cytoplasmic inclusion bodies, dispersion of the cytoplasmic polyribosomes, mitochondrial swelling, and cellular extrusion by 24 h after injection. The next two studies were performed using tissue cultures from mature avian auditory sensory epithelium. Cultured cells were labeled using vital fluorescent markers, and levels of intracellular calcium and reactive oxygen species within hair cells were studied following aminoglycoside exposure. We identified a dose-dependent increase in the levels of intracellular calcium, which was blocked by an inhibitor of voltage-gated calcium channels. We also found that levels of reactive oxygen species in hair cells greatly increased after exposure to gentamicin, and this response was blocked by two different antioxidants. These studies serve to identify key cellular and molecular changes in hair cells in response to ototoxic drugs. Further study of these processes may lead to a better understanding of how ototoxicity is induced and to potential preventative interventions. PMID- 10842610 TI - Intratympanic therapy for Meniere's disease. High-concentration gentamicin with round-window protection. AB - Many therapeutic options exist for the management of patients with Meniere's disease. In the last few years, the use of intratympanic gentamicin has been investigated as an alternative treatment to vestibular nerve section or labyrinthectomy. In humans, the concentration of gentamicin used for intratympanic treatment of vertigo ranges from 10 mg/mL to 40 mg/mL, and the number of doses from 2 to 14, with a total administered amount between 6 and 2.400 mg. Here lower doses of gentamicin were used, usually had the lowest incidence of hearing loss, but more injections were needed to ablate vestibular function. The purpose of this study was to evaluate the acute and chronic ototoxic effects of intratympanic high-concentration gentamicin after having obliterated the round-window niche with connective tissue in 11 subjects' ears with Meniere's disease. Intratympanic gentamicin was administered according to a predetermined and fixed schedule consisting of two doses of 0.5 mL gentamicin solution, injected once a week with a drug concentration of 80 mg/mL. The total dose of gentamicin was < or = 80 mg. The charts of the patients were surveyed in accordance with the 1995 AAO-HNS guidelines. Three patients had recurrence of vertigo between 3 and 6 months after the second injection and went on to one additional dose of gentamicin. At 2 years follow-up, 10 patients (91%) had complete and 1 (9%) substantial control of vertigo; 3 subjects (27%) had hearing decreased. Tinnitus disappeared or decreased in 3 patients (27%); eight subjects (73%) reported their aural pressure abolished or decreased. The present study demonstrates that in patients with Meniere's disease, 0.5 mL gentamicin solution, with a concentration of 80 mg/mL (total dose < or = 80 mg), injected intratympanically once a week after having obliterated the round-window niche, permits complete or substantial control of vertigo in two-thirds of cases after two doses and in all subjects after three doses. This vertigo control rate is compared to that observed after vestibular nerve section. Hearing results are not different from those with natural control, with endolymphatic sac surgery, and with vestibular nerve section. PMID- 10842611 TI - Cochlear effects of mesna application into the middle ear. AB - Mesna (sodium 2-mercapto-ethane sulphonate) belongs to a class of thiol compounds that produce mucolysis by disrupting the disulphide bonds of the mucus polypeptide chains. The registered indications of mesna include the treatment of pathologies of the respiratory tract and, in oncology, the prevention of toxic lesions of the urinary tract by antineoplastic agents. In the E.N.T. Clinic of the University of Parma, it has been found that mesna can be used to facilitate the dissection of the various tissue layers in any surgical procedure. One of these indications is surgical treatment of cholesteatoma, which is mainly composed by keratin, a protein rich is disulphide bonds that are easily disrupted by mesna. The aim of this study was to evaluate the toxicity of mesna application into the middle ear on the cochlear anatomy and physiology. Three groups of guinea pigs were used as subjects. Mesna solution (10 or 20%) was applied in one ear, while the opposite ear received a placebo (saline solution). Toxicity of mesna was assessed by means of transmission electron microscopy (TEM), scanning electron microscopy (SEM), and auditory brain-stem response (ABR). TEM and SEM did not show any toxic effect on cochlear morphology. There were no differences in ABR thresholds and wave III amplitude and latency between mesna-treated and control ears. PMID- 10842612 TI - The cochleovestibular system/ototoxicity/clinical issues. AB - It is recommended that the experiences of basic science with ototoxicity be applied to the development of specific clinical strategies for the treatment of the cochleovestibular complaints of hearing loss, tinnitus, vertigo, and ear blockage. The treatment goals are to maintain/improve function, namely, hearing and balance, as well as to control aberrant auditory and vestibular phenomena, for example, tinnitus and vertigo originating within the peripheral and/or central cochleovestibular system. It is hypothesized that a "common" neurochemical basis for "toxicity" exists, whatever the primary toxic etiology. It is also recommended that the neurochemical mechanisms of neuroprotection and apoptosis be considered for investigations of ototoxicity. Furthermore, it is suggested that ototoxicities are reflected in clinical manifestations of sensory cochleovestibular complaints of hearing loss, tinnitus, and vertigo, and affect complaints of stress, pain, and neuropsychiatric disorders. PMID- 10842613 TI - Health implications of global electrification. PMID- 10842614 TI - Network construction of first aid for electrical injury in the east China area of China Power. AB - Electrical injury is an area of burn medicine that requires specialized treatment. Injury rates have reached 6.56% in 1990, compared to only 1% prior to 1980. In recent years, the morbidity has steadily increased due to the rapid development of the electrical power industry and the popularization of electrical installations. Electrical accidents cause severe damage to muscle and tissue, which makes treatment of these accident patients quite challenging. Therefore, it is very important to promote the widespread use of a first aid network within the electrical power system while strengthening safety measures within the industry to reduce morbidity and mortality among the electrically injured. To date, similar networks have been established in East China, Central China, North China, and Shandong within the six major electrical power industries. We hope to minimize the injury of workers through appropriate supervision, improved safety measures, and improved safeguarding of equipment. PMID- 10842615 TI - Survey of electrical burns in Philippine General Hospital. AB - A total of 211 patients were admitted at the UP-PGH Burn Unit in 1995. Sixty eight (32.2%) patients suffered burn injuries secondary to electrical injury. In the period from July to December 1995, a total of 28 patients were admitted because of burns caused by electricity. Twenty-six (92%) patients sustained injury secondary to high tension (> 1000 V) electrical current and 2 (7.1%) secondary to low tension (< 1000 V) electrical current, specifically 220 V, which were both self-inflicted. Twenty-seven (96%) patients were males. Injury was work related in 18 patients, nonwork-related in 8, and self-inflicted in 2. Sixteen (57%) patients suffered injuries on a rooftop, 11 (39%) patients on the ground, and 1 (3.5%) patient on top of an electric post. A total of 22 operations were performed on 28 patients, including 7 extremity amputations, which makes an amputation rate of 25%. There were 5 deaths secondary to overwhelming wound sepsis, making an overall mortality rate of 21.7% and an operative mortality rate of 22.7%. The average length of hospital stay was 17.2 days. PMID- 10842616 TI - Thermoacoustic energy effects in electrical arcs. AB - Electrical arcs commonly occur in electrical injury incidents. Historically, safe work distances from an energized surface along with personal barrier protection have been employee safety strategies used to minimize electrical arc hazard exposures. Here, the two-dimensional computational simulation of an electrical arc explosion is reported using color graphics to depict the temperature and acoustic force propagation across the geometry of a hypothetical workroom during a time from 0 to 50 ms after the arc initiation. The theoretical results are compared to the experimental findings of staged tests involving a mannequin worker monitored for electrical current flow, temperature, and pressure, and reported data regarding neurologic injury thresholds. This report demonstrates a credible link between electrical explosions and the risk for pressure (acoustic) wave trauma. Our ultimate goal is to protect workers through the design and implementation of preventive strategies that properly account for all electrical arc-induced hazards, including electrical, thermal, and acoustic effects. PMID- 10842617 TI - Common reasons for electric injuries and preventive measures in China's electric power industry. PMID- 10842618 TI - Electrical injury through the eyes of professional electricians. AB - This paper presents the first results of an attitudinal survey that was conducted among professional electricians in order to explore their knowledge and beliefs related to occupational electrical injury. Four hundred eighty-one out of 1200 questionnaires were returned and analyzed. The presented statistical and qualitative data reflect electricians' personal experiences with electrical injury, their communication patterns around electrical trauma, their understanding of possible electrical injury sequelae, their safety beliefs and attitudes towards occupational dangers, and the reasons for their occupational choice. We expect that the results of this study will enhance our understanding of the psychological profile, environment, and culture of electrical workers. The collected data may also help to identify those at risk for poor outcome after electrical injury and determine a new set of risk factors to be taken into account by medical professionals, social workers, and union/utility training officers. PMID- 10842619 TI - Treatment of severe electrical burns. AB - In summary, electrical injury is a typical type of burn injury with a unique mechanism of systemic and local damage. Resuscitation is more complex in critical patients than that of thermal injury. Local management is more difficult, which could be eventually accomplished by sequential procedures of debridement and operation. Although the modern trend of performing early debridement and operation might be promising, traditional experiences are still being used in clinical care. Progress in clinical management has brought about an improvement in therapeutic outcome, resulting in a decrease of amputation rate and reduced disability. Some typical types of electrical injury demand special treatment, which might prove to be effective. Close observation and timely treatment of the injury itself and related complications will be lifesaving in caring for critical electrical injury. Some special therapeutic measures would be indicated and might be fruitful in proving their efficacy. New measures aimed at theoretical problems will be promising if electroporation could be effectively treated. PMID- 10842620 TI - Electrical injuries in Canadian burn care. Identification of unsolved problems. AB - Over the past decade, the Firefighters' Burn Treatment Unit of the University of Alberta Hospital in Edmonton, Alberta, Canada, has treated 1399 inpatients suffering from thermal injury. Regional burn care is provided in a 10-bed intensive care unit with 18 plastic surgery reconstructive beds for a large referral region of central and northern Alberta, portions of the Northwest Territories, and neighboring provinces of British Columbia and Saskatchewan. Of the total burn inpatients during this period, 74 electrical injuries were treated (5.3% of all admissions): 71 were males (95.9%) and 3 females (4.1%). The mean age of all patients was 33.9 +/- 12.6 years (range 1-67). Compared to our general population of thermally injured patients, those with electrical injuries had smaller injuries [9.9 +/- 12.9% TBSA (range 1-65) versus 15.1 +/- 10.1], shorter length of hospitalization [18.6 +/- 7.3 days (range 1-80) versus 26.2 +/- 0.8], and substantially lower mortality once reaching the hospital (0% versus 4%). Electrical injuries were classified as flash in 30 cases, contact in 42 cases, and lightning in 2 cases; 74.3% of injuries occurred during work-related activities. A total of 118 operative procedures were performed during the acute admission (1.6 procedures per patient), including 19 amputations: 12 in the upper and 7 in the lower extremity. The mean time of amputation was 9.3 +/- 5.3 days after admission. In contact injuries of the upper extremity, 14 patients suffered amputations or neurologic injury that required reconstruction with free tissue transfers and nerve grafts. Long-term functional outcome of these patients using sensory testing, the Jebsen-Taylor hand function test, and wound coverage has revealed that these patients have substantial persistent sensory impairment of their upper extremities postinjury despite reconstruction, although many remain active and functional with acceptable wound coverage. Based on our analysis of electrical injury as it presents to one typical Canadian burn unit, our patients suffer limb loss on a delayed basis, which leads to substantial morbidity. Reconstruction of the upper extremity with microsurgical techniques after profound electrical injury has provided acceptable coverage, but in many instances is associated with poor or marginal sensory recovery limiting reemployment options for patients with upper extremity electrical burns. Further understanding of the cellular biology of delayed tissue loss after electric injury would offer the potential for reduction in amputation rate and improvement in functional outcome and overall morbidity. PMID- 10842621 TI - A clinical analysis of 836 cases with electric injury. AB - Eight hundred thirty-six patients with electric injury were admitted to Shanghai Power Hospital from 1972 to 1998. The majority of electric injury patients were young or middle-aged industrial workers from both cities and rural areas. These patients were classified into three categories depending on the type of electric injury: (1) electric shock; (2) electric contact burn injury; (3) electric arc burn. In our study of wound management, we focused on three areas: first, the choice of operation time in electric burn injury; second, the methods of wound management on the injured extremities, especially the hands and upper limbs; third, functional site wounds that included some exposed blood vessels. Nerves and tendons were covered and repaired by various flaps. Through the analysis of 836 electric injury cases, we summarized our experience of treating electric injury with the purpose of increasing our knowledge in the area of electric injury prevention and treatment. PMID- 10842622 TI - Functional reconstruction following electrical injury. AB - Worldwide, high voltage electrical injury continues to cause significant morbidity, disability, and mortality despite improvements in electrical safety. Joule heating and cell membrane disruption are two mechanisms important in understanding the pathophysiology of electrical injury. The degree of tissue damage is often more extensive than initially perceived on clinical exam. Using modern reconstructive techniques, the functional outcome of electrical injury victims can be improved. The type of reconstruction selected for each injury can be selected from a number of options from the reconstructive ladder. Donor site considerations, risks of the surgery, team experience, and patient preference are important factors in this selection. PMID- 10842623 TI - Wound management in electrical injuries. PMID- 10842624 TI - The use of allografts in the management of exposed calvarial electrical burn wounds of the skull. AB - Deep burns of the scalp and skull are often caused by high voltage electrical injuries. In an early stage, protection and preservation of the skull is important. Covering the wound with a full-circulatory skin flap or a myocutaneous free flap is a reasonable selection, but is not our first choice. In our experience, preliminary use of glycerol-preserved allograft wound dressing significantly improved the development of granulation tissue to perform an autograft at a later stage. With these allografts, there is a significantly shortened healing time compared with the results found in the literature. PMID- 10842625 TI - Experience of urgent reconstruction of electrical injuries. AB - Although there have been great advances in the treatment of electrical injuries in the last 20 years, the extremity loss ratio in electrical injuries remains at an unacceptably high level. The primary reason for this is the progressive tissue necrosis and enlargement of the necrosis in the wound. The goal in this study is to examine possible ways to break the necrotic malignancy circle and save the form and function of damaged extremities. As a result of systematic experimental and clinical research, a comprehensive urgent reconstruction alternative for electrical injuries has been proposed. The alternative includes the following principles: debriding the wound as early as possible after injury; preserving as much as possible the vital tissue structures, such as nerve, vessels, joints, tendons, and bone, even when they have undergone devitalization or local necrosis; transplanting these vital tissues during the first surgery if the functional reconstruction requires; nourishing the wound bed by covering with tissue flaps that have rich blood supply; improving flap survival by continuous irrigations with a compound medicine beneath the flaps for a 24- to 72-hour period after surgery; providing general treatment with vasoactive agents and antibiotics. This paper summarizes our experience of using this method in the treatment of 105 electrical injury patients (a total of 309 wounds) in the time period from 1 January 1986 to 31 December 1996. Satisfying results were obtained, with the extremity loss ratio decreasing to 7% as compared to 41.5% during the 10 years preceding 1984 at the same hospital. Thus, the urgent comprehensive reconstruction alternative presented here is an effective and workable method to manage electrical injuries. PMID- 10842626 TI - Experience of the treatment of severe electric burns on special parts of the body. AB - The treatment in these eight cases with severe electric burns on special parts of the body was successful. Our experience can be summarized as follows: (1) The role of the immediate measures including resuscitation at the scene of the accident cannot be understated. If the patient presents with a complex injury such as open pneumothorax, first aid should be provided immediately and then the patient should be transferred to a specialized treatment center. (2) General conditions such as the presence of shock, water-electrolyte balance, renal function, and others should be continuously monitored. (3) Antibiotics should be suitably administered and combined with antianaerobic drugs. (4) For different wound sites, different plans of treatment, including various immediate and delayed procedures, could be appropriate. In life-threatening cases such as exposed carotid artery, perforative injury of the chest wall, spinal cord damage, and others, first-stage repair using skin flap or myocutaneous flap must be performed after early debridement. For other wound sites, such as oral area and tongue, eye socket, and penis, second-stage reconstruction may be more suitable for better cosmetic appearance and function. (5) Adequate nutritional supply and early treatment of anemia may expedite wound healing. PMID- 10842627 TI - High voltage electrical injury leading to a delayed onset tetraplegia, with recovery. AB - High voltage electrical injury can cause considerable damage to the central nervous system. Delayed spinal cord injury is uncommon, usually incomplete, and comprises predominantly motor fallout. The injury can be progressive, with only a few patients showing partial improvement. We present a case report of a 20-year old male who had a delayed onset spinal cord injury after a high voltage electrical injury. The symptoms started several days postburn with an ascending paralysis, leading to tetraplegia. Gradual recovery became evident at 3 months after the accident, starting with his arms and later showing partial recovery of his lower limbs. The mechanisms of injury and pathophysiology to the spinal cord are poorly understood. Possible explanations for the delayed onset of neurologic complications are given. The exact reasons for the delayed, ascending paralysis and the mechanism of recovery still need further investigation. PMID- 10842628 TI - Wound repair of severe electrical injuries in the head. PMID- 10842629 TI - The neuropsychological effects of electrical injury. New insights. PMID- 10842630 TI - Understanding the cellular and molecular basis of electrical shock. PMID- 10842631 TI - Issues in modeling thermal alterations in tissues. AB - Thermal injury in living tissues is commonly modeled as a rate process in which cell death is interpreted to occur as a function of a single kinetic process. Experimental data indicate that multiple rate processes govern the manifestation of injury and that these processes may act over a broad spectrum of time domains. Injury is typically computed as a dimensionless function (omega) of the temperature time history via an Arrhenius relationship to which numerical values are assigned based on defined threshold levels of damage. However, important issues central to calculation and interpretation of the omega function remain to be defined. These issues include the following: how is temperature identified in time and space within a tissue exposed to thermal stress; what is the biophysical and physiological meaning of a quantitative value for omega; how can omega be quantified in an experimental system; how should omega be scaled between graded levels of injury; and what are the differences in injury kinetics between unit volume- and unit surface area-governed processes of energy deposition into tissue to cause thermal stress? This paper addresses these issues with the goal of defining a more rigorous and comprehensive standard for modeling thermal injury in tissues. PMID- 10842632 TI - Pathophysiology and treatment of fibroproliferative disorders following thermal injury. PMID- 10842633 TI - Changes in skin structure and electrical properties following high voltage exposure. AB - Human skin experiences extensive alterations when exposed to a strong electric field. In high voltage electric shocks, tissues in the current path undergo large scale changes; the stratum corneum (SC), the outermost layer of the skin, loses its barrier function against ionic transport. The primary mechanism of electric field interaction with the skin is the creation of aqueous pathways, which increase the permeability of the skin to ions and macromolecules. Changes in skin structure and electrical properties were characterized by measuring the transport of fluorescent molecules in an in vitro preparation of human cadaver skin in a permeation chamber. Application of high voltage pulses (Uskia > 150 V, tau pulse = 1 ms) decreased the skin resistance per unit area by three orders of magnitude from its initial value of 100 k omega-cm-2. The resistance reached a quasi-steady state after about 20 pulses. After the cessation of pulsing, Rskin recovered immediately to about 4 k omega-cm-2 and remained low for 2 h thereafter. Transport of fluorescent molecules by high voltage pulses occurs in highly localized transport regions (LTRs), with diameters not exceeding 100 microns for short pulses (tau pulse < 5 ms). PMID- 10842634 TI - Dynamics of membrane sealing in transient electropermeabilization of skeletal muscle membranes. AB - Large supraphysiologic transmembrane electrical potentials are known to alter the molecular organization of the bilayer lipid component of cell membranes, leading to ionic permeabilization or "electroporation". Typically, membrane electroporation is followed by several orders of magnitude increases in electrical conductance and diffusive permeability to low-molecular-weight solutes. Electroporation may be transient or stable depending on whether the membrane eventually seals or remains permeabilized. Factors that control sealing have not been well characterized. This paper describes the kinetics of membrane sealing following electroporation by pulses over a range of supraphysiologic potentials. The increase in membrane conductance is highly nonlinear during a 440-mV, 4-ms pulse and reaches two orders of magnitude greater than baseline. Electroporation and relaxation sealing kinetics are quite different, reflecting a significant hysteresis effect. Thus, it appears that the magnitude and duration of the field pulse are important factors in sealing. PMID- 10842635 TI - Biological effects of electric shock and heat denaturation and oxidation of molecules, membranes, and cellular functions. AB - Direct exposure of cells in suspension to intense electric pulses is known to produce damages to cell membranes and supramolecular organizations of cells, and denaturation of macromolecules, much like injuries and tears seen in electric trauma patients. Thus, the system has been used as a laboratory model for investigating the biochemical basis of electric injury. An intense electric pulse can produce two major effects on cells--one caused by the field, or the electric potential, and the other by current, or the electric energy. The field-induced transmembrane potential can produce electro-conformational changes of ion channels and ion pumps and, when the potential exceeds the dielectric strength of the cell membrane (approximately 500 mV for a pulse width of a few ms), electro conformational damages and electroporations of membrane proteins and lipid bilayers. These events lead to passage of electric current through the membrane porated cells and to heating of cell membranes and cytoplasmic contents. The subsequent denaturation of cell membranes and cytoplasmic macromolecules brings about many complex biochemical reactions, including oxidation of proteins and lipids. The combined effects may cripple the cells beyond repair. This communication will focus on the thermal effects of electric shock. After a brief review of the current state of knowledge on thermal denaturation of soluble enzymes and muscle proteins, this paper will describe experiments on the thermal denaturation of cellular components and functions, such as nucleosomes, and the electron transport chain and ATP synthetic enzymes of the mitochondrial inner membranes. Data will show that lipid peroxidation and the subsequent loss of the energy-transducing ability of the cells may occur even at moderate temperatures between 40 degrees C and 45 degrees C. However, lipid peroxidation may be prevented with reducing reagents such as mercaptoethanol, dithiothreitol, and ascorbic acid. Reactivation of denatured cellular proteins and functions may also be possible and a strategy for doing so is discussed. PMID- 10842636 TI - The use of collagen-GAG membranes in reconstructive surgery. AB - Porous collagen-glycosaminoglycan (PCG) membranes with a porous silicone elastomer coating have been useful as a scaffold for dermal replacement in burn victims. Critical physicochemical parameters of these membranes include pore size, cross-link density, the percentage of glycosaminoglycan, and the degree of banding of the collagen. These factors govern the immunobiological response. Optimizing these parameters can reduce inflammation, scarring, and contraction of wounds grafted with PCG membranes. PCG membranes are currently commercially manufactured (Integra, Integra Life Sciences, New Jersey) and available for clinical use. Because clinical outcomes have improved using these membranes for burn wound coverage, other skin reconstruction problems including scar resurfacing, keloids, treatment of donor sites, and treatment of chronic wounds can be considered as potential applications. This manuscript illustrates our early experience using Integra as a CG membrane for dermal replacement in reconstructive surgery. Our results indicate that CG membranes can lead to improved compliance and appearance compared to a meshed graft and may be sequentially placed in multiple layers to correct contour deformities. Also, in one case, we observed that, if placed on a wound bed with embedded skin epithelial cells, the PCG promotes epithelialization through the PCG matrix. The use of this material results in a supple integument with many similarities to normal skin. PMID- 10842637 TI - Changes in electroporation thresholds of lipid membranes by surfactants and peptides. AB - This article reviews recent work from our laboratory that explores how chemical additives may alter the threshold of electroporation of synthetic lipid bilayers. The addition of the nonionic block copolymeric surfactant, poloxamer 188 (P188), at a concentration of 1 mM increased the electroporation thresholds of planar lipid bilayer membranes made of azolectin. For a 10-microsecond rectangular pulse, P188-treated membranes were found to have a statistically higher threshold voltage, longer latency time to rupture, and lower postpulse conductance. Addition of the nonionic surfactant, octaethyleneglycol-mono-n-dodecyl-ether (C12E8), decreased the electroporation threshold of bilayer membranes made of 1 palmitoyl-2-oleoyl-phosphatidylcholine (POPC) by 10-40% for 10-microsecond- to 10 s-duration pulses, in a concentration-dependent manner over concentrations ranging from 0.1 to 10 mM. Postpulse membrane conductance also increased. The opposite effects of the two surfactants on electroporation thresholds may result from their very different structures, which would encourage different modes of surfactant-lipid interactions. To examine protein-lipid interactions and their effects on the electroporation threshold, the effects of a channel-forming polypeptide, gramicidin D (gD), was studied on membrane conductance and electroporation threshold. Electroporation thresholds for 15-ms pulses were unaffected by addition of gramicidin to POPC at a peptide:lipid concentration estimated to be 1:10,000, but increased significantly at ratios of 1:500 and 1:15, while membrane conductance increased monotonically with peptide concentration. A micropipette aspiration technique was applied to giant unilamellar POPC vesicles to measure changes in the membrane physical properties. When gD was added to give an estimated peptide:lipid ratio of 1:15, the membrane area expansivity modulus increased, indicating that the increase in electroporation threshold is correlated with a change in membrane stiffness. Thus, these findings demonstrate that surfactants or peptides can mediate the electroporation threshold of lipid bilayers. PMID- 10842638 TI - Pharmaceutical therapies for sealing of permeabilized cell membranes in electrical injuries. AB - Several years ago, we proposed that loss of cell membrane structural integrity by electroporation is a substantial cause of tissue necrosis in victims of electrical trauma. Specifically, this involves the permeabilization of the lipid bilayer by thermal and electrical forces. We further suggested that certain mild surfactants in low concentration could induce sealing of permeabilized lipid bilayers and salvage of cells that had not been extensively heat-damaged. Successful restoration of membrane transport properties using the surfactant poloxamer 188 was reported in 1992. The purpose of this study is to further examine the response of electroporated rat skeletal muscle membranes to poloxamer 188 (P188) therapy by direct assay of membrane transport properties. Experimental evidence accumulated to date suggests that P188 is effective in sealing permeabilized cell membranes both in vitro and in vivo. PMID- 10842639 TI - Oxidative cell membrane alteration. Evidence for surfactant-mediated sealing. AB - Exposure to very intense ionizing irradiation produces acute tissue sequelae including inflammation, pain, and swelling that often results in tissue fibrosis and/or necrosis. Acute tissue necrosis occurs in hours when sufficiently rapid damage to membrane lipids and proteins leads to altered membrane structure, disrupting the vital electrochemical diffusion barrier necessary for cell survival. This damage mechanism is thought to underlie the interphase death of lethally irradiated postmitotic cells such as neurons, but it has also been implicated in the rapid cell death of lymphocytes and acute vascular changes due to capillary epithelium dysfunction. It is not known whether sealing of radiation permeabilized cell membranes will prolong survival of lethally irradiated cells or perhaps lead to repair of damaged nucleic acids. The purpose of this study is to begin to address the first question. PMID- 10842640 TI - Radiotracers for imaging electroporation. PMID- 10842641 TI - Study on the distribution pattern of skin temperature in normal Chinese and detection of the depth of early burn wound by infrared thermography. AB - OBJECTIVE: Conduct basic research of the distribution pattern of skin temperature and estimate the wound depth in normal Chinese in the early stage of burn injury by using technical thermography. METHODS: (1) Two hundred twenty-three volunteers from the electrical power system were randomly chosen: 138 males and 85 females, mean age 42.5 +/- 8.8, and mean weight 65.37 +/- 9.7 kg. (2) Thermovision 470 Pro camera of AGEMA Corporation, Sweden, was used. (3) A total of 29,882 values of skin temperature from all over the body were classified, compared, and statistically analyzed. RESULTS: The difference in temperature on the y-axis of the body is relatively significant, from 0 to 9 degrees C. The difference in temperature on the x-axis of the body is relatively invariable. CONCLUSIONS: The depth of early burn wound can be detected by using infrared thermography, which is an objective, fast, non-touching, and noninvasive method. PMID- 10842642 TI - Conductivity mapping of biological tissue using diffusion MRI. PMID- 10842643 TI - Using engineering and assistive technologies for rehabilitation after electrical trauma. AB - A framework is presented for judging when, how, and why rehabilitation engineering and its related assistive technologies are appropriate interventions following electrically induced trauma or burns. Instead of relying on the World Health Organization's medically based classification scheme of "Impairment, Disability, and Handicap," this newer framework is built on a rational demarcation proposed by the National Center for Medical Rehabilitation and Research at the U.S. National Institutes of Health. This latter client-centered framework encompasses pathophysiology, impairment, functional limitations, "disability", and societal limitations. This framework is well suited to handle the varied sequelae of electrical trauma and burn injuries and provides guidance towards the most effective use of traditional rehabilitation interventions and of assistive technologies. For electrical injuries, rehabilitative technologies can be classified as those promoting job accommodations (i.e., that help an individual return to active employment, albeit possibly in a different role) or as aids to the other activities of daily living (ADLs) that provide an enhanced quality of life to the individual with disability. While the traditional rehabilitative focus has been on return-to-work, especially among professional tradesmen, a more productive rehabilitative effort in some cases may occur through psychosocial adjustments achieved via effective technological interventions that enhance ADLs. PMID- 10842644 TI - Early repair treatment of electrical burns and recovery of tendons and nerves. Report of 194 operations. AB - A new approach to treatment of electrically burned tendons and nerves by primarily covering with skin or myocutaneous flaps is recommended. Between 1964 and 1989, 194 operations using the new approach were performed on 147 patients, of which 179 operations were successful and only 15 procedures resulted in infection. Among 42 cases involving the wrist, the rate of functional recovery of the electrically burned tendons was 97.6%; the rate of recovery in 21 cases of electrically burned peripheral nerves was 80.9%. PMID- 10842645 TI - Management and coordination of postacute medical care for electrical trauma survivors. AB - The clinical spectrum of electrical injury ranges from the absence of any external physical signs to severe multiple trauma. Reported neuropsychiatric sequelae can vary from vague complaints, which may seem unrelated to the injury in their occurrence over time or by their apparent severity, to sequelae consistent with brain injury accompanying an electrical trauma. In this report, a case study and discussion are presented on the management and coordination of post-acute care of an electrical trauma survivor. Expertise and a multidisciplinary team are essential to cohesive patient care. Patient monitoring for progressive changes and prompt intervention are needed to address the potential difficulties experienced by trauma survivors as they rehabilitate to return to their work and their activities of daily living. PMID- 10842646 TI - Strategies in disability management. Corporate disability management programs implemented at the work site. AB - Managers are challenged to demonstrate all programs as economically essential to the business, generating an appreciable return on investment. Further challenge exists to blend and integrate clinical and business objectives in program development. Disability management programs must be viewed as economically essential to the financial success of the business to assure management support for clinical interventions and return-to-work strategies essential for a successful program. This paper discusses a disability management program integrating clinical and business goals and objectives in return-to-work strategies to effect positive clinical, social-cultural, and business results. Clinical, educational, social, and economic challenges in the development, implementation, and continued management of a disability program at a large corporation with multiple global work sites are defined. Continued discussion addresses the effective clinical interventions and educational strategies utilized successfully within the workplace environment in response to each defined challenge. A multiple disciplinary team approach, clinical and business outcome measures, and quality assurance indicators are discussed as major program components. This article discusses a successful program approach focusing on business process and methodology. These parameters are used to link resources to strategy, developing a product for implementing and managing a program demonstrating economic value added through effective clinical medical case management. PMID- 10842647 TI - Life after electrical injury. Risk factors for psychiatric sequelae. AB - Long-term cognitive and emotional deficits have been commonly reported in electrical injury (EI) survivors. However, it remains undetermined what factors may lead to the development of such effects in some patients and not in others. In this study, we hypothesized that certain elements of subjective EI experience may predict specific psychiatric sequelae. A group of 73 post-acute EI patients were included in this retrospective study. Statistical associations were examined between major psychiatric diagnoses (posttraumatic stress disorder and major depression) and such EI descriptors as having experienced "no-let-go" or having been knocked away on contact, as well as loss of consciousness or altered states of consciousness at the scene of the accident (including amnesia for the event). The study results will help physicians determine which patients may be at increased risk of developing psychiatric symptoms and address these issues as part of their total rehabilitation plan. PMID- 10842648 TI - Future development of electrical injury research and treatment. PMID- 10842649 TI - The metabolic syndrome X. PMID- 10842650 TI - Etiology of the metabolic syndrome: potential role of insulin resistance, leptin resistance, and other players. AB - Obesity and Type 2 diabetes are now major public health issues in developed nations and have reached epidemic proportions in many developing nations, as well as disadvantaged groups in developed countries, e.g., Mexican-Americans, African Americans, and Australian Aborigines. These groups all show hyperinsulinemia and insulin resistance, which have been demonstrated to be future predictors of Type 2 diabetes and have also been suggested as key factors in the etiology of the Metabolic Syndrome. It is now increasingly recognized that Type 2 diabetes is part of a cluster of cardiovascular disease (CVD) risk factors comprising the Metabolic Syndrome. This group is at very high risk of atherosclerosis because each of the risk factors in the Metabolic Syndrome cluster in its own right is an important CVD risk factor. They also contribute cumulatively to atherosclerosis. A key strategy in reducing macrovascular disease lies in the better understanding of the Metabolic Syndrome--glucose intolerance, hypertension, hyperlipidemia, and central obesity. Although it has been suggested that hyperinsulinemia/insulin resistance is the central etiological factor for the Metabolic Syndrome, epidemiological data do not support the idea that this can account for all of the cluster abnormalities. We have animal and human data suggesting that hyperleptinemia rather than, or synergistically with, hyperinsulinemia may play a central role in the genesis of the CVD risk factor cluster that constitutes the syndrome. Studies in Psammomys obesus (the Israeli sand rat) suggest hyperinsulinemia/insulin resistance is an early metabolic lesion in the development of obesity and Type 2 diabetes. This animal also develops other features of the Metabolic Syndrome, making it an excellent model to investigate etiology. Psammomys, when placed on an ad libitum laboratory diet, develops hyperinsulinemia, insulin resistance, impaired glucose tolerance, diabetes, and dyslipidemia. It also develops hyperleptinemia and leptin insensitivity, and hyperleptinemia is correlated with insulin resistance independent of changes in body weight. It is likely that a similar sequence occurs in the transition from the prediabetic state to Type 2 diabetes in humans. More recently, other potential players in the etiology of the Metabolic Syndrome have been suggested including endothelial dysfunction and acetylation-stimulating protein (ASP). It has been suggested that endothelial dysfunction may be an antecedent for both Type 2 diabetes and the Metabolic Syndrome. In addition, ASP is a serious new candidate for an important role in insulin resistance. The ASP pathway plays a critical role in fatty acid metabolism and storage, and it has been suggested that ineffective storage of fatty acids by adipocytes due to a defect in the ASP pathway may lead to insulin resistance and Type 2 diabetes. PMID- 10842651 TI - Insulin resistance: a chicken that has come to roost. AB - Insulin-mediated glucose disposal varies approximately 10-fold in apparently healthy human beings. Insulin (I)-resistant individuals can remain glucose tolerant if the pancreas compensates for this defect by secreting large amounts of I. Type 2 diabetes develops when I-resistant persons cannot sustain this state of compensatory hyperinsulinemia (increases I). However, the ability of increases I to prevent decompensation of glucose tolerance is a mixed blessing, and the combination of I resistance and increases I predisposes such individuals to develop a series of abnormalities that increase risk of coronary heart disease (CHD). Given the health-related consequences of I resistance and increases I, it has been suggested that a "thrifty" genotype exists that favored evolutionary survival by enhancing I secretion and thereby promoting energy accumulation. An alternative view is that conservation of muscle mass was necessary for survival, and that muscle I resistance was the "thrifty" genotype. This latter hypothesis is more consistent with current data, and there is evidence of a genetic basis for I resistance. In either case, there is little question as to the importance of I resistance and related abnormalities in diseases of Western civilization. However, the strength of the association between I resistance and its consequences varies in magnitude, and it is necessary to emphasize that development of a clinical end-point will vary as a function of (1) degree of I resistance; (2) "closeness" of I resistance to the end-point; and (3) the ability to compensate for the effects of I resistance. I resistance is a physiological characteristic, genetically determined, that helped primitive humans to survive. It is greatly aggravated by obesity and physical inactivity, and represents a modern scourge. PMID- 10842652 TI - Interaction between aging and syndrome X: new insights on the pathophysiology of fat distribution. AB - Increased fat mass (FM), and in particular a specific increase in visceral fat (VF), may account for the age-associated decrease in insulin action and the development of Syndrome X. Utilizing chronic caloric restriction (CR) with aging in a rodent model, we dissociated the effects of VF and FM, and demonstrated that the decrease in VF accumulation was sufficient to prevent the marked decrease in hepatic insulin action. This suggests that the typical increase in VF with aging, rather than aging per se, determines hepatic insulin resistance. To directly assess the role of VF, we studied rats after surgical removal of VF or sham operation. Surgical extraction of VF (which accounts for approximately 10% of total fat) improved hepatic insulin action by more than twofold. We studied the role of fat-derived peptides in the regulation of body composition and insulin action. While VF extraction resulted in decreased gene expression for leptin and TNF-alpha in the subcutaneous adipose, administration of leptin selectively decreased visceral fat (approximately 60%), and enhanced the action of insulin on inhibiting hepatic glucose production (approximately 80%). Thus, the cause-effect relationship between the age-related increase in VF and the decrease in hepatic insulin action may involve the failure of leptin to "cross talk" with other fat depots to regulate fat distribution. PMID- 10842653 TI - Mechanisms for hyperglycemia in the metabolic syndrome. The key role of beta-cell dysfunction. AB - Hyperglycemia in Type 2 diabetes represents a steady-state re-regulation of plasma glucose to a higher-than-normal level after an overnight fast. The underlying pathophysiology represents an interaction between impaired beta-cell function and peripheral and hepatic insulin resistance which leads to abnormal hepatic glucose production. Subjects with the Metabolic Syndrome are at an increased risk for Type 2 diabetes and often have one or both of these disorders present even when glucose tolerance is normal. Thus, sophisticated measures of beta-cell function and insulin sensitivity demonstrate a high frequency in populations characterized as having a high prevalence of atherosclerosis, central obesity, hypertension, and dyslipidemia with or without impaired glucose tolerance. Hyperglycemia compensates for the impairment of beta-cell function and therefore, in our view, the beta-cell is the critical factor in its development. Hyperinsulinemia, a curvilinear compensation for insulin resistance that is closely correlated with central adiposity, is another important predictor of hyperglycemia. In a Japanese-American population followed for five years, impaired beta-cell function was present at baseline and preceded the accumulation of intraabdominal fat in those who developed Type 2 diabetes five years later. This interaction between these two pathophysiologic abnormalities in this sequence supports the hypothesis that beta-cell dysfunction contributes to the development of central adiposity by reduced CNS insulin signaling. PMID- 10842654 TI - Role of the sympathetic adrenal system in the pathogenesis of the insulin resistance syndrome. AB - The pathophysiology of the various manifestations of Syndrome X has been poorly understood. A possible mechanism involves stimulation of the sympathetic nervous system (SNS). Insulin plays an important role in the relationship between dietary intake and SNS activity. Because insulin-mediated glucose uptake in central hypothalamic neurons regulates SNS activity in response to dietary intake, a hypothesis was developed that links the hyperinsulinemia of obesity to sympathetic stimulation, the latter exerting a prohypertensive effect mediated by the kidney, the heart, and the vasculature. Evidence in support of this hypothesis has been obtained from the Normative Aging Study (NAS) in which a relationship between insulin (and glucose) and the SNS, and between insulin and SNS activity and blood pressure was demonstrated. The characteristic dyslipidemia in NAS subjects, moreover, was related to insulin and epinephrine. As reported in other studies, insulin level was directly associated with low HDL and high triglyceride levels. An independent inverse association was also noted between urinary epinephrine excretion and lipid levels: high epinephrine excretion rates were associated with high HDL and low triglyceride levels and, conversely, low epinephrine excretion was associated with low HDL and high triglycerides. In the NAS, therefore, increased SNS activity contributes to hypertension while diminished adrenal medullary activity contributes to the low HDL and high triglyceride levels commonly seen in association with hypertension. PMID- 10842655 TI - Mechanisms of hypertension and kidney disease in obesity. AB - Abnormal kidney function is an important cause as well as a consequence of obesity. Excess renal sodium reabsorption, probably in the loop of Henle, and a hypertensive shift of pressure natriuresis play a major role in initiating increased blood pressure associated with weight gain. The mechanisms responsible for increased sodium reabsorption and altered pressure natriuresis in obesity include activation of the renin-angiotension and sympathetic nervous systems, and physical compression of the kidneys due to accumulation of intrarenal fat and extracellular matrix. Sympathetic activation may be mediated, in part, by elevated circulating leptin and interactions with neuropeptides in the hypothalamus. Renal remodeling and extracellular matrix proliferation likely involve complex interactions between intrarenal physical forces, neurohumoral factors, and local growth factors and cytokines. Although glomerular hyperfiltration and increased arterial pressure help to compensate for increased renal tubular reabsorption in the early phases of obesity, these changes also increase glomerular capillary wall stress which, along with activation of neurohumoral systems and increased lipids and glucose intolerance, cause glomerular cell proliferation, matrix accumulation, and eventually glomerulosclerosis and loss of nephron function in the early phases of obesity. This creates a slowly developing vicious cycle that requires additional increases in arterial pressure to maintain sodium balance and therefore makes effective antihypertensive therapy more difficult. Because obesity is the main cause of Type 2 diabetes and an important cause of human essential hypertension, it seems likely that obesity is also one of the most important risk factors for end-stage renal disease. PMID- 10842656 TI - Chronic angiotensin II infusion promotes atherogenesis in low density lipoprotein receptor -/- mice. AB - The purpose of this study was to determine the effect of chronic angiotensin II (AngII) infusion on the severity of the atherogenic process in low density lipoprotein (LDL) receptor -/- mice with established lesions. LDL receptor -/- mice receiving a diet enriched in cholesterol, saturated fat, and cholate, were infused with saline or AngII (500 ng/kg/min) for 28 days. Systolic blood pressure increased in LDL receptor -/- mice following 7 days of AngII infusion, followed by a decline to baseline levels at 28 days, despite continued AngII infusion. Serum cholesterol was not influenced by AngII infusion in LDL receptor -/- mice; however, serum triglyceride concentrations were reduced significantly in LDL receptor -/- mice receiving AngII. The percent of intimal surface area covered by lesion was not increased in LDL receptor -/- mice receiving AngII; however, the content of cholesterol (esterified and unesterified) in lesions of the arch, thoracic, and abdominal aorta was significantly increased in those mice infused with AngII. Of note, in 20% of the LDL receptor -/- mice receiving AngII, large aneurysms were found in the abdominal aorta. Aneurysms appeared as breaks in the media and surrounding tissue of the vessel wall, encompassing amorphous and acellular masses with patches of thrombotic material. These results demonstrate that chronic infusion of AngII promotes the atherogenic processes in LDL receptor -/- mice, manifest as increases in lesion cholesterol content. Effects of AngII to promote atherogenesis were apparent at doses which did not markedly elevate systolic pressure. Importantly, infusion of AngII in LDL receptor -/- mice resulted in the development of aortic aneurysms. PMID- 10842657 TI - Insulin signaling and action in fat cells: associations with insulin resistance and type 2 diabetes. AB - Adipose tissue only accounts for a relatively small proportion (< 10%) of the peripheral glucose utilization in response to insulin. However, the fat cells may still play an important role in insulin resistance and Syndrome X through, for instance, its endocrine functions (production of leptin, TNF alpha, PAI-1, etc.) and involvement in lipid metabolism (FFA release and hydrolysis of triglycerides). The fat cells are also highly sensitive to insulin and may thus be used to elucidate molecular mechanisms for insulin resistance in man. Examinations of the intracellular signaling mechanisms for insulin in fat cells from individuals with Type 2 diabetes revealed markedly lower insulin-stimulated PI3-kinase activity. This was due to a pronounced reduction in the cellular expression of the docking protein, IRS 1, whereas expression of IRS 2 was normal. However, IRS 2-associated PI3-kinase activity was only approximately one-third of that found to be associated with IRS 1 in normal cells. Downstream activation and serine phosphorylation of PKB/Akt by insulin were also markedly reduced in Type 2 diabetes. Furthermore, the dose-response curve for this effect of insulin was similar to that for glucose transport in both normal and Type 2 diabetic cells. Thus, these data show that both PI3-kinase and PKB activation by insulin are markedly reduced in Type 2 diabetes. We also examined whether an attenuated activation of PI3-kinase by insulin can be seen in non-diabetic insulin-resistant states. Approximately 30% of healthy subjects with at least two first-degree relatives with Type 2 diabetes exhibited perturbations in IRS-1 expression and signaling. These individuals were characterized by insulin resistance as well as other markers of Syndrome X. Thus, impaired IRS-1 expression and downstream signaling events in fat cells in response to insulin are associated with insulin resistance and Syndrome X. PMID- 10842658 TI - Cross talk between adipocytes and their precursors: relationships with adipose tissue development and blood pressure. AB - Adipose tissue is an important source of angiotensinogen (AT). A possible involvement of increased plasma AT has been proposed in hypertension in obese patients, but growing evidence suggests also that the local renin-angiotensinogen system (RAS), giving rise to angiotensin II (AngII), may act as a distinct system from the plasma RAS. AngII stimulates in vitro the production and release of prostacyclin from adipocytes which in turn promotes the differentiation of precursor cells into new adipocytes (Darimont et al. 1994, Endocrinology 138: 1092-1096). Cross talk between adipocytes and precursor cells has been validated ex vivo and in vivo as (1) AngII stimulates specifically prostacyclin production, (2) both carbacyclin, a stable analogue of prostacyclin, and AngII promote the formation of new fat cells, and (3) AT (-/-) mice, which have decreased blood pressure (Tanimoto et al. 1994, J. Biol. Chem. 269: 31334-31336), exhibit both hypertrophy and hypoplasia of adipose tissue compared to wild-type mice (collaboration with Prof. A. Fukamizu, Tsukuba University, Japan). Altogether the data are consistent with an autocrine/paracrine mechanism implicating AT, AngII, and prostacyclin in adipose tissue development and suggest a new role for local AngII in addition to that of systemic AngII in blood pressure. Hormonal regulation of AT production from adipose tissue is also discussed. PMID- 10842659 TI - Role of PPAR gamma in regulating adipocyte differentiation and insulin-responsive glucose uptake. AB - Adipocyte differentiation is regulated by at least two families of transcription factors, CCAAT/enhancer binding proteins (C/EBPs) and peroxisome proliferator activated receptors (PPARs). Induction of PPAR gamma gene transcription during the differentiation of preadipocytes into adipocytes in vitro occurs following an initial phase of cell proliferation and requires a direct involvement of C/EBP beta, C/EBP delta, and glucocorticoids. Ectopic expression of PPAR gamma in non adipogenic, Swiss 3T3 fibroblasts promotes their conversion into adipocytes as indicated by the accumulation of lipid droplets and the induction of C/EBP alpha, aP2, insulin-responsive aminopeptidase (IRAP), and glucose transporter 4 (GLUT4) expression. These PPAR gamma-expressing Swiss cells also exhibit a high level of insulin-responsive glucose uptake that is comparable to that expressed in 3T3-L1 adipocytes. In contrast, PPAR gamma-expressing NIH-3T3 fibroblasts, despite similar lipid accumulation, adipocyte morphology, and aP2 expression, do not synthesize C/EBP alpha and fail to acquire insulin sensitivity. In Swiss 3T3 cells ectopically expressing PPAR gamma, the development of insulin-responsive glucose uptake correlates with C/EBP alpha expression. Furthermore, ectopic expression of C/EBP alpha in NIH-3T3 cells induces PPAR gamma expression and adipogenesis, but also restores insulin-sensitive glucose transport. These results suggest that although PPAR gamma is sufficient to trigger the adipogenic program, C/EBP alpha is required for establishment of insulin-sensitive glucose transport in adipocytes. PMID- 10842660 TI - Molecular mechanism of metabolic syndrome X: contribution of adipocytokines adipocyte-derived bioactive substances. AB - Syndrome X is a clinical syndrome in which multiple risks cluster in an individual, and it is a common basis of vascular disease in the industrial countries. The molecular basis of Syndrome X, however, has not been elucidated. We have analyzed body fat distribution using CT scan and have shown that people who have accumulated intra-abdominal visceral fat frequently have multiple risks and vascular diseases. Thus, "visceral fat syndrome" is a clinical entity compatible with Syndrome X. To clarify the molecular mechanism of the disorders in visceral fat syndrome, we analyzed the expressed genes in adipose tissue by a large-scale random sequencing. Unexpectedly, visceral fat expressed a variety of the genes for secretory proteins including various bioactive substances; we designated them adipocytokines. One of them, plasminogen activator inhibitor-1, was overproduced in accumulated visceral fat and might contribute to the development of vascular disease. We have also cloned a novel adipose-specific gene named adiponectin. Adiponectin is a collagen-like plasma protein which has an inhibitory effect on proliferation of vascular smooth muscle cells; its plasma levels are paradoxically decreased in obesity. Adipocytokines may play important roles in the development of the disorders in Syndrome X. PMID- 10842661 TI - On the control of lipolysis in adipocytes. AB - The lipolytic reaction in adipocytes is one of the most important reactions in the management of bodily energy reserves, and dysregulation of this reaction may contribute to the symptoms of Type 2 diabetes mellitus. Yet, progress on resolving the molecular details of this reaction has been relatively slow. However, recent developments at the molecular level begin to paint a clearer picture of lipolysis and point to a number of unanswered questions. While HSL has long been known to be the rate-limiting enzyme of lipolysis, the mechanism by which HSL attacks the droplet lipids is not yet firmly established. Certainly, the immunocytochemical evidence showing the movement of HSL to the lipid droplet upon stimulation leaves little doubt that this translocation is a key aspect of the lipolytic reaction, but whether or not HSL phosphorylation contributes to the translocation, and at which site(s), is as yet unresolved. It will be important to establish whether there is an activation step in addition to the translocation reaction. The participation of perilipin A is indicated by the findings that this protein can protect neutral lipids within droplets from hydrolysis, but active participation in the lipolytic reaction is yet to be proved. Again, it will be important to determine whether mutations of serine residues of PKA phosphorylation sites of perilipins prevent lipolysis, and whether such modifications abolish the physical changes in the droplet surfaces that accompany lipolysis. PMID- 10842662 TI - Signaling pathways mediating insulin-stimulated glucose transport. AB - A major action of insulin is to accelerate the rate of uptake of sugar into muscle and adipose cells following a meal. The biochemical mechanism by which this is accomplished has been a subject of intense experimentation, although elucidation of the pathways has remained elusive. In recent years, numerous signaling molecules and cascades modulated by insulin have been identified, although few have been definitively established as important to the metabolic actions of the hormone. An exception to this is the lipid kinase phosphatidylinositide 3'-kinase, which, under many conditions, appears absolutely required for insulin to stimulate hexose uptake into adipocytes. Akt/PKB, a serine/threonine protein kinase activated by insulin in a phosphatidylinositide 3'-kinase-dependent manner, has been implicated as a critical mediator of insulin's actions on metabolism and cell survival. Nonetheless, Akt/PKB's role in many insulin effects, particularly accelerated glucose transport, remains controversial. Interestingly, soluble analogues of ceramide antagonize both insulin's activation of Akt/PKB as well as its stimulation of glucose transport, consistent with a causal relationship between the two. PMID- 10842663 TI - Nutrient modulation of cellular insulin action. AB - Abundant evidence supports a crucial role for dietary factors in the induction and maintenance of insulin resistance. At the cellular and tissue level, the availability of substrates for cellular energy production may play an important role in metabolic regulation and, in particular, in determining the response to insulin stimulation. The infusion of amino acids or fatty acids decreases insulin stimulated glucose disposal in vivo; sustained hyperglycemia also induces insulin resistance. To determine whether nutrients directly affect insulin signaling, we have evaluated the impact of fatty acids, amino acids, and activation of the hexosamine pathway on insulin signaling in both cultured cells and animal models. We demonstrate that fatty acids and amino acids inhibit early post-receptor steps in insulin action, including tyrosine phosphorylation of insulin receptor substrate (IRS) proteins and activation of phosphatidylinositol 3-kinase (PI3 kinase), both in vitro and in several in vivo models. Similarly, activation of the hexosamine pathway by infusion of glucosamine also reduces insulin-stimulated phosphorylation of IRS proteins, activation of PI3-kinase, and activation of glycogen synthase. These data suggest that nutrients directly modulate insulin signaling, perhaps via common pathways, and thus contribute to cellular insulin resistance. PMID- 10842664 TI - Role of PC-1 in the etiology of insulin resistance. AB - Defects in insulin receptor tyrosine kinase activity have been demonstrated in tissues from insulin resistant subjects, but mutations in the insulin receptor gene are rare. Therefore, other molecules that are capable of modulating the insulin receptor most likely play a major role in insulin resistance. In cultured fibroblasts from an insulin resistant patient with Type 2 diabetes, we first identified membrane glycoprotein PC-1 as an inhibitor of the insulin receptor tyrosine kinase activity. PC-1 is overexpressed in fibroblasts from other insulin resistant subjects, both with and without Type 2 diabetes. PC-1 is a large class II exoprotein whose function is unknown. Studies in muscle and fat of insulin resistant subjects two primary tissues for insulin activation, reveal that elevated levels of PC-1 are inversely correlated with decreased insulin action both in vivo and in vitro. Transfection and expression of PC-1 in cultured cells demonstrate that overexpression of PC-1 produces impairments in insulin receptor tyrosine kinase activity, and the subsequent cellular responses to insulin. These studies indicate, therefore, that PC-1 is a major factor in the etiology of insulin resistance, and is a potential new therapeutic target for anti-diabetic therapy. PMID- 10842665 TI - Nutritionally induced insulin resistance and receptor defect leading to beta-cell failure in animal models. AB - Animals with genetically or nutritionally induced insulin resistance and Type 2 diabetes comprise two groups: those with resilient beta-cells, e.g., ob/ob mice or fa/fa rats, capable of longstanding compensatory insulin hypersecretion and those with labile beta-cells in which the secretion pressure leads to beta-cell degranulation and apoptosis, e.g., db/db mice and Psammomys gerbils (sand rats). Psammomys features low insulin receptor density; on a relatively high energy diet it becomes hyperinsulinemic and hyperglycemic. In hyperinsulinemic clamp the hepatic glucose production is only partially suppressed by insulin, even in the normoglycemic state. The capacity of insulin to activate muscle and liver receptor tyrosine kinase is nearly abolished. GLUT4 content and mRNA are markedly reduced. Hyperinsulinemia was also demonstrated to inhibit insulin signaling and glucose transport in several other species. Among the factors affecting the insulin signaling pathway, phosphorylation of serine/threonine appears to be the prominent cause of receptor malfunction as inferred from the finding of overexpression of PKC epsilon isoforms in the muscle and liver of Psammomys. The insulin resistance syndrome progressing in animals with labile beta-cells to overt diabetes and beta-cell failure is a "thrifty gene" characteristic. This is probably also true for human populations emerging from food scarcity into nutritional affluence, inappropriate for their metabolic capacity. Thus, the nutritionally induced hyperinsulinemia, associated with PKC epsilon activation may be looked upon from the molecular point of view as "PKC epsilon overexpression syndrome." PMID- 10842667 TI - Neuroendocrine regulation of nutrient partitioning. AB - Leptin is a satiety factor which acts within the hypothalamus to decrease the levels of several neuropeptides stimulating food intake (among them, neuropeptide Y [NPY]), while increasing those that inhibit food intake. These effects of leptin bring about decreased body weight. In vivo, leptin potentiates basal and insulin-stimulated glucose utilization, presumably its oxidation, and decreases fat storage. Leptin increases sympathetic-mediated energy dissipation, and the expression of uncoupling proteins-1, -2, and -3. In peripheral tissues (muscles, adipose, others), leptin decreases triglyceride content by increasing fatty acid oxidation, decreasing the activity/expression of esterification and lipogenic enzymes, and favoring lipolysis. It decreases the lipogenic activity of insulin. Ultimately, leptin depletes fat stores and promotes leanness. NPY, taken as one example of what an orexigenic agent may produce, increases food intake and body weight. It favors fat storage in adipose tissue by stimulating lipogenic activity. It decreases glucose utilization by muscles, making more glucose carbon available for lipogenesis. Effects of NPY result from vagus nerve-mediated hyperinsulinemia and overactivity of the hypothalamo-pituitary-adrenal axis. Thus, NPY favors fat stores, and ultimately obesity. Glucocorticoids are necessary for NPY effects to occur, because central administration of the neuropeptide in adrenalectomized animals is ineffective. Glucocorticoids also have genuine effects when administered centrally to normal rats. They increase the hypothalamic content of NPY and decrease that of CRH. This double neuro peptidic change stimulates food intake, insulin output, adipose tissue storage ability, decreases the expression of uncoupling proteins-1 and -3, and increases body weight. Body weight homeostasis appears to require a finely tuned regulation of both leptin and glucocorticoids, with their respective opposite effects. PMID- 10842666 TI - Paradoxical phosphorylation of skeletal muscle glycogen synthase by in vivo insulin in very lean young adult rhesus monkeys. AB - Calorie restriction (CR) has previously been shown to unexpectedly induce a reversal of in vivo insulin action (phosphorylation instead of dephosphorylation) on skeletal muscle glycogen synthase (GS) in four out of six long-term calorie restricted (CR) monkeys. The purpose of the present study was to determine whether this increase in Ka (concentration of glucose 6-phosphate [G6P] at which GS activity is half-maximal) during insulin is also present in very lean (VL) young adult monkeys maintained on a controlled feeding regimen. Muscle samples from 10 VL monkeys (10 +/- 2% body fat; 7 years old) were obtained before and during a euglycemic hyperinsulinemic clamp and the Ka was determined and compared to the Ka of two other groups of monkeys, one matched in age but fully ad libitum (AL)-fed (n = 9.8 +/- 1 years old, 20 +/- 3% body fat, p = 0.01 vs. VL monkeys), and the other our previously described weight-clamped long-term CR monkeys (n = 6.20 +/- 1 years old, 21 +/- 2% body fat, p = 0.01 vs. VL monkeys). All of the AL monkeys had the expected decrease in Ka with insulin; however, similar to the 4 out of 6 CR monkeys, 7 out of 10 VL monkeys had an increase in Ka with insulin. The 11 monkeys with an increase in Ka (+Ka) (7 VL + 4 CR) were compared to the 14 monkeys with a decrease in Ka with insulin (-Ka) (3 VL + 2 CR + 9 AL). The +Ka monkeys had lower basal Ka (p = 0.0001), higher basal GS fractional activity (p = 0.0003), lower basal G6P content (p = 0.002), lower glycogen phosphorylase fractional activity (p = 0.01), and lower whole-body insulin-mediated glucose disposal rate (p < 0.05) than the -Ka monkeys. We conclude that the condition of steady-state restrained calorie intake (as in the CR monkeys and in the controlled feeding VL monkeys) produces the paradoxical action of in vivo insulin to phosphorylate muscle GS, and raises the possibility that the presence of the unusual response to insulin may serve as a marker in calorie-restrained individuals for the genotype of obesity, insulin resistance and/or Type 2 diabetes. PMID- 10842668 TI - Molecular pathology in the obese spontaneous hypertensive Koletsky rat: a model of syndrome X. AB - The SHROB rat is a unique strain with genetic obesity, hypertriglyceridemia, hyperinsulinemia, renal disease with proteinuria, and genetically determined hypertension, characteristics paralleling human Syndrome X. The obese phenotype results from a single homozygous recessive trait, designated faK, and is allelic with the Zucker fatty trait (fa), but of distinct origin. The faK mutation is a premature stop codon in the extracellular domain of the leptin receptor, resulting in a natural receptor knockout. The SHROB are glucose intolerant compared to heterozygous or wild-type SHR, but retain fasting euglycemia even on a high sucrose diet, suggesting that diabetes requires polygenic interaction with additional modifier genes. Insulin-stimulated phosphorylation of tyrosine residues on the insulin receptor and on the associated docking protein IRS-1 are reduced in skeletal muscle and liver compared to SHR, due mainly to diminished expression of insulin receptor and IRS-1 proteins. Despite multiple metabolic derangements and severe insulin resistance, hypertension is not exacerbated in SHROB compared to SHR. Thus, insulin resistance and hypertension are independent in this model. Increased activity of the sympathetic nervous system may be a common factor leading by separate pathways to hypertension and to insulin resistance. We studied the chronic effects of sympathetic inhibition with moxonidine on glucose metabolism in SHROB. Moxonidine (8 mg/kg/day), a selective I1-imidazoline receptor agonist, not only reduced blood pressure but also ameliorated glucose intolerance. Moxonidine reduced fasting insulin by 47% and plasma free fatty acids by 30%. Moxonidine enhanced expression and insulin stimulated phosphorylation of IRS-1 in skeletal muscle by 74 and 27%, respectively. Thus, central sympatholytic therapy not only counters hypertension but also insulin resistance, glucose tolerance, and hyperlipidemia in the SHROB model of Syndrome X. PMID- 10842669 TI - Transgenic mice lacking white fat: models for understanding human lipoatrophic diabetes. AB - The human disease lipoatrophic (or lipodystrophic) diabetes is a rare syndrome in which a deficiency of adipose tissue is associated with Type 2 diabetes. This disease is an interesting contrast to the usual situation in which diabetes is associated with obesity, an excess of fat. Aside from obesity, patients with lipodystrophic diabetes have the other features associated with Metabolic Syndrome X, including hypertension and dyslipidemia. The contrast between diabetes with a lack of fat and diabetes with an excess of fat provides an opportunity to study the mechanisms causing Type 2 diabetes and its complications. Recently, three laboratories have produced transgenic mice that are deficient in white adipose tissue. These mice have insulin resistance and other features of lipoatrophic diabetes, and are a faithful model for the human disease. Here we review the different murine models of fat ablation and compare the murine and human diseases, addressing the questions: Is the lack of fat causative of the diabetes, and if so by what mechanism? How could the other clinical features be explained mechanistically? And finally, what can be gleaned about insight into treatment options? PMID- 10842670 TI - Hypothalamic origin of the metabolic syndrome X. AB - The conspicuous similarities between Cushing's syndrome and the Metabolic Syndrome X open up the possibility that hypercortisolemia is involved also in the latter. Salivary cortisol is possible to measure during undisturbed conditions including perceived stressful events during everyday life. Such measurements clearly show that normally regulated cortisol secretion is associated with excellent health in anthropometric, metabolic, and hemodynamic variables. Upon perceived stress cortisol secretion is increased and followed by the Metabolic Syndrome X (insulin resistance, abdominal obesity, elevated lipids, and blood pressure). In a minor part of the population a defect, "burned-out" cortisol secretion, occurs with decreased sex steroid and growth hormone secretions, and strong, consistent associations with the Metabolic Syndrome X. Psychosocial and socioeconomic handicaps with tendencies to abuse and depressive-anxious mood changes are consistently associated. The feedback control of cortisol secretion by central glucocorticoid receptors (GR) is blunted, and the function of the GR is abnormal. This corresponds to a polymorphism early in the GR gene locus, which is also associated with abdominal obesity and insulin resistance and is found in 14% of the Swedish male population. We suggest that the Metabolic Syndrome X is due to a discretely elevated cortisol secretion, discoverable during reactions to perceived stress in everyday life. This is based on environmental factors and expressed with different impact depending on genetic susceptibility. PMID- 10842671 TI - Excessive portal venous supply of long-chain free fatty acids to the liver, leading to hypothalamus-pituitary-adrenal-axis and sympathetic activation as a key to the development of syndrome X. A proposed concept for the induction of syndrome X. PMID- 10842672 TI - Pharmacological interference with hepatic glucose production. PMID- 10842673 TI - The SHROB model of syndrome X: effects of excess dietary sucrose. PMID- 10842674 TI - Very low density lipoprotein overproduction is maximally expressed in early insulin resistant states. PMID- 10842675 TI - Metabolic basis of low high density lipoprotein in subjects with impaired glucose tolerance. PMID- 10842676 TI - Early pubertal development and overweight in girls. PMID- 10842677 TI - A computer program for clinical and investigative work related to the metabolic syndrome. PMID- 10842679 TI - Habits of the heart. PMID- 10842680 TI - Mothering: letting go of the past ideal and valuing the real. PMID- 10842678 TI - Leptin does not attenuate the hyperphagia induced by 2-deoxy-D-glucose. PMID- 10842681 TI - Constructing daily routines: a qualitative examination of mothers with young children with disabilities. AB - This qualitative research study explored the daily routines that mothers construct in response to the emerging self-care skills of their young children with disabilities. Over 2 months, data were collected from naturalistic observations and in-depth interviews of six mothers and their children. Ecocultural theory was used to examine the influence of ecological constraints and cultural values on the construction of routines. The daily occupations of these families were shaped by the simultaneous process of accommodating to ecocultural influences and anticipating future possibilities. A mother's vision for her child's future also played a pivotal role in determining whether emerging skills would be reinforced as a part of the home routines. PMID- 10842682 TI - Mothers with disabilities: in their own voice. AB - OBJECTIVES: The purpose of this study was to gain an understanding of the personal experience of women with disabilities engaged in the occupation of mothering and their perceptions of their interpersonal environment, including interaction with family, professional caregivers, and the community. METHOD: In this qualitative study, in-depth interviews were conducted with a diverse sample (n = 8) of mothers with disabilities or chronic illnesses to uncover the nature of their mothering experience. Data were analyzed for themes with the constant comparison approach of grounded theory. RESULTS: The quality of the participants' experience varied with the degree of perceived similarity or dissimilarity with other mothers and acceptance of these differences. In addition, their perception of the supportive or nonsupportive nature of their interpersonal environment had an impact on their mothering experience. CONCLUSION: The results suggest that mothers with disabilities tend to value the importance of performing maternal occupations according to a perceived culturally common way, although self acceptance of their own unique differences in performing these occupations facilitates maternal engagement as well. PMID- 10842683 TI - The orchestration of occupation: the dance of mothers. AB - OBJECTIVES: This article describes the relationship of mothers' orchestration of daily occupations, the specialized maternal work of parenting a child with a disability, and the mother's subjective well-being. METHOD: Mothers' daily occupations and subjective well-being were studied using multiple in-depth interviews, participant observation of a day's round of occupations, and scales of well-being. Data were treated to a recursive analysis, which included theoretical notes generated during transcriptions that identified important themes and additional points of inquiry, line-by-line coding of transcripts, and theoretical sorting of codes and regrouping, recoding. To account for patterns in the data, a relational analysis was conducted that included the generation of metaphors. RESULTS: Emergent findings of this analysis identified the mothers' guiding occupational motif and eight processes of orchestration in their daily routines. The occupational motif, the embrace of paradox, directed the mother's orchestration of daily occupations. The orchestration processes included planning, organizing, balancing, anticipating, interpreting, forecasting, perspective shifting, and meaning making. Examples illustrate the maternally driven and child-sensitive nature of these processes. CONCLUSION: In their daily rounds, the mothers studied were attentive to the manner and method with which they interacted with their children to produce child-contingent occupations commensurate with their values of being a good mother. Using these orchestration processes, mothers made sense of their past, designed their present, and planned for their future within their daily occupational rounds for themselves and family members. PMID- 10842684 TI - Toward an understanding of mothering: a comparison of two motherhood stages. AB - OBJECTIVE: This study sought to understand the tasks and activities involved in the caring and nurturing work of mothering, a common and important occupation for many women. METHOD: In-depth, semistructured individual interviews were conducted with 40 mothers: 20 mothers of preschool-age children and 20 mothers of young adults. In addition, participants completed a questionnaire describing the tasks and activities that they currently engage in when caring for their children. RESULTS: The activities involved in mothering are different for the mothers at the two stages of mothering examined. The mothers of preschool-age children are very involved in caretaking tasks and meeting the basic needs of the child, whereas the mothers of young adults are involved in emotional and supportive type activities. Mothers at both stages are involved in caring and nurturing work but this work evolves and changes as children mature. CONCLUSION: These findings extend our awareness of mothering and the tasks and activities involved in the occupation of mothering at both the preschool stage and young-adult stage. PMID- 10842685 TI - Maternal management of the home as a developmental play space for infants and toddlers. AB - OBJECTIVE: One purpose of this qualitative study was to describe the work done by mothers as they manage the spaces and objects of the home to support the development of infants and toddlers at play. METHOD: Eighteen mother-infant dyads participated in the study. Data were collected via monthly in-home videotaping of infants and monthly interviews with mothers, from 1 to 18 months of infant age. Data were analyzed with a grounded theory approach and computer-assisted video and text analysis. RESULTS: The results describe the everyday tasks of mothers of infants and toddlers, such as selecting commercial toys and household objects for play, positioning infants for play, maintaining and making play objects available, furnishing the home with child care equipment, controlling infant access to the spaces of the home, and monitoring for safety. CONCLUSION: This description contributes to our understanding of maternal work, infant and toddler development in context, co-occupations, and the neglected spatial dimension of occupation. PMID- 10842686 TI - Adaptive strategies of mothers with children with attention deficit hyperactivity disorder: enfolding and unfolding occupations. AB - OBJECTIVE: To describe adaptive strategies of time use among mothers with children with attention deficit hyperactivity disorder (ADHD) and, in particular, the adaptive strategies of unfolding occupations that were commonly used by these mothers. METHOD: In-depth interviews were conducted with 17 families with children with ADHD. Parents were asked to describe their family's daily schedule, routines, and how occupations were performed. Data were analyzed using the grounded theory approach. FINDINGS: Mothers used three adaptive strategies: enfolding occupations, temporal unfolding of occupations, and unfolding occupations by inclusion. Enfolding occupations means performing more than one occupation at a time. Unfolding occupations means taking chunks of activities or occupations out of previously established sequences of enfolded occupations to be performed at a different time (temporal unfolding) or by a different person (unfolding by inclusion). The child's special needs and the availability of financial and human resources influenced the selection of adaptive strategy. Using these adaptive strategies enabled the mothers to care for their children. However, using enfolding or temporal unfolding meant that mothers had less time for other occupations. CONCLUSION: The benefits and limitations of the adaptive strategies of enfolding and unfolding occupations suggest a need to study time use in different contexts. Before suggesting use of these adaptive strategies by parents of children with special needs, therapists need to be aware of the family's financial and human resources. PMID- 10842687 TI - Mothering young children with disabilities in a challenging urban environment. AB - OBJECTIVE: This study examined the occupations of mothering very young children with physical disabilities. METHOD: Two single, urban, Black mothers in their mid 20s were interviewed with a semistructured protocol. The audiotaped, transcribed data were analyzed with a phenomenological method. RESULTS: The context of the challenging urban environment was a constant influence that shaped the participants' occupations of mothering. The major over-arching theme derived from the data was that mothering was "what I got to do" in relation to their particular child. Two sub-themes were identified: (a) mothering as caring and (b) the impact of social supports on the occupations of mothering. CONCLUSION: To plan meaningful, effective interventions, occupational therapy practitioners need to understand the context in which mothering occupations occur and to ensure that mothers' caring occupations and social support needs are addressed in the therapeutic partnership. PMID- 10842688 TI - Time use and leisure occupations of young offenders. AB - OBJECTIVE: The overall goal of this study was to understand the time use, including leisure occupations, of a sample of young offenders in Melbourne, Australia. METHOD: This study investigated how 37 probationary young offenders (from 13-18 years of age) spent their wakeful time during 1 week. The study used a combination of the Experience Sampling Method and interviewing. Participants were beeped 60 times over 7 days and, each time, they were asked to complete a questionnaire about the occupations in which they were engaged. Each participant was interviewed both before and after the Experience Sampling Method data collection about their everyday lives, including their leisure occupations. Data were analyzed qualitatively and quantitatively. RESULTS: The young offenders reported that they were engaged in leisure occupations, predominantly passive, 57% of the times beeped, and in personal care occupations 21% of the times beeped. Only 10% of the times beeped did they report being engaged in productive occupations such as education or employment. The time spent in passive leisure occupations was 30% higher than for the average Australian adolescent. Leaving school and lack of financial and human resources contributed to the high percentage of engagement in passive leisure occupations. CONCLUSION: Findings from this study help us to understand the relationship between use of time and social well-being, particularly the nature of time use of young offenders, and will help to inform occupational therapy practices with such groups. PMID- 10842689 TI - Relationship between performance on tests of basic visual functions and visual perceptual processing in persons after brain injury. AB - OBJECTIVE: In this correlational study of adults receiving occupational therapy who sustained a cerebrovascular accident (CVA), the relationship between basic visual functions (including acuity, visual field deficits, oculomotor skills, and visual attention or scanning) and higher level visual-perceptual processing skills (e.g., visual closure and figure-ground discrimination) was investigated. METHOD: Thirty adults who sustained CVA and 20 adults without a history of CVA were given a basic visual function screening and the Motor-Free Visual Perception Test (MVPT). Scores on the vision screening and the MVPT were correlated statistically. RESULTS: A Pearson product-moment correlation analysis produced a correlation of r = .75 between vision screening scores and scores from the MVPT. CONCLUSION: These results suggest that a positive relation exists between basic visual functions and visual-perceptual processing skills. Further, the results suggest that evaluation of visual-perceptual processing skills must begin with assessment of basic visual functions so that the influence of these basic visual functions on performance in more complex tests can be taken into consideration. PMID- 10842690 TI - Use of the student-oriented learning outline in research education. PMID- 10842691 TI - The role of occupational therapy in a developmental technology model. PMID- 10842693 TI - Levels of research competence across the profession of occupational therapy. PMID- 10842692 TI - Communicating with clients, family members, and colleagues about research evidence. PMID- 10842694 TI - Cerebral amyloidosis in prion diseases. PMID- 10842695 TI - Amyloid beta peptide: a century of discoveries. PMID- 10842696 TI - Serum amyloid A: from fibril to function. Current status. PMID- 10842697 TI - AL amyloidosis: the last 30 years. PMID- 10842698 TI - Hereditary amyloidosis: some words on the history and present status of our eminent subject. PMID- 10842699 TI - Beta 2-microglobulin amyloid. PMID- 10842700 TI - Endocrine amyloid--a subject of increasing interest for the next century. PMID- 10842701 TI - The relation of proteoglycans, serum amyloid P and apo E to amyloidosis current status, 2000. PMID- 10842702 TI - Expression of SAA and amyloidogenesis in congenic mice of CE/J and C57BL/6 strains. AB - Inbred strains of mice display different susceptibilities to experimental induction of reactive amyloidosis. CBA/J, C57BL/6, and ICR are among the most susceptible strains, while CE/J mice appear to be totally resistant. In contrast to amyloidogenic strains which express two major acute phase serum amyloid A proteins (SAA1 and SAA2), CE/J mice produce only a single isoform, designated SAA2.2. Studies indicate that CE/J x CBA/J hybrid mice expressing both SAA2.2 and SAA1.1/SAA2.1 are amyloid resistant, and this has led to the hypothesis that SAA2.2 may protect mice against amyloid formation even in the presence of fibrillogenic SAA1.1. We have tested this hypothesis in mice derived from CE/J and C57BL/6 strains. CE/J mice were mated with C57BL/6 mice to produce F1 hybrids. Congenic mice were then produced by backcrossing each successive generation to C57BL/6 mice. Representative mice from F2 and F3 generations were analyzed to determine SAA genotype and susceptibility to amyloid induction by repeated casein injections. All F2 and F3 mice examined, including those which carried the SAA2.2 gene, developed extensive splenic AA amyloid. Expression of SAA2.2 in mice testing positive for the SAA2.2 gene was confirmed by sequence analysis of HDL-associated SAA proteins. These results demonstrate that the presence of SAA2.2 is not sufficient to protect CE/J x C57BL/6 hybrid mice from amyloid development. This is consistent with our observation that macrophage cultures, derived from C57BL/6, CBA/J, or CE/J mice, undergo amyloid deposition when treated with SAA1.1 alone or with equal amounts of SAA1.1 and SAA2.2, but show no deposition when treated solely with SAA2.2. We conclude from these studies that while SAA2.2 is non-fibrillogenic, its physical presence is not sufficient for protection against amyloid formation. PMID- 10842703 TI - Amyloid formation in the rat: adenoviral expression of mouse serum amyloid A proteins. AB - Serum amyloid A (SAA) proteins are acute-phase apolipoproteins that are associated with high-density lipoprotein (HDL) particles: SAA proteins are precursors to secondary amyloid fibril proteins and under certain conditions of chronic or recurrent inflammation these proteins are deposited as amyloid fibrils. Of two isotypes found in mouse, SAA1.1 and SAA2.1, only SAA1.1 is deposited into amyloid. The CE/J mouse is unique, in that the only isoform identified is a hybrid between SAA1.1 and SAA2.1 and the mouse does not show amyloid deposition. In the rat, a deletion in the SAA1/SAA2 gene is associated with the absence of protein in the plasma and subsequently no amyloid deposition is detected. We have generated adenoviral vectors to study the expression of SAA proteins on HDL metabolism and amyloid formation. Injection of SAA viruses into rats resulted in expression of the mouse SAA proteins in the plasma with specific association of the SAA with HDL particles. The induction of SAA proteins was comparable to that seen in mice presented with the inflammatory agent, bacterial lipopolysaccharide (LPS). Adenoviral induced SAA levels were maintained for up to several weeks without a significant decrease in SAA expression. Injection of rats with the mouse SAA1.1 adenoviral vector, followed by amyloid enhancing factor (AEF) and silver nitrate resulted in the deposition of amyloid fibrils in the spleen. After 2 weeks, amyloid could be detected in other tissues, including the heart, liver, kidneys and lungs. When animals were injected with null or the SAA2.2 virus no amyloid was detected. These studies demonstrate that the inability of the rat to develop AA amyloid is due to the lack of synthesizing an amyloidogenic SAA protein. Furthermore, the expression of the adenoviral SAA protein from the liver and incorporation onto HDL particles further supports the hypothesis that AA amyloid is derived from circulating SAA protein. The ease of use of the adenoviral vectors and the rat provide an excellent model to study the function of SAA proteins. PMID- 10842704 TI - Analysis of plasma cell clonality in localized AL amyloidosis. AB - AL amyloidosis is characterized by fibrillar tissue deposits composed of monoclonal immunoglobulin light chains(IgLs). It has been speculated that clonal expansion of plasma cells may occur locally and produce amyloidogenic IgLs. Both immunohistochemistry and molecular genetics are useful for examining plasma cell clonality from paraffin-embedded tissue sections, which are easy to obtain. We evaluated plasma cell clonality in 16 biopsy cases of localized AL amyloidosis using these two methods. A clonal excess of plasma cells was detected in 6 (37.5%) cases immunohistochemically, in 10 (62.5%) cases molecularly, and in 13 (81.3%) cases by at least one of the two methods. These results support local synthesis of the light chain proteins in localized AL amyloidosis. PMID- 10842706 TI - AL 612, a glycosylated lambda light chain of subgroup I. PMID- 10842705 TI - A novel variant of transthyretin (Glu89Lys) associated with familial amyloidotic polyneuropathy. AB - We detected a point mutation in the transthyretin (TTR) gene associated with familial amyloidotic polyneuropathy (FAP) in a 57-year old male presenting with sensorimotor polyneuropathy, severe autonomic dysfunction and cardiomyopathy using a non-isotopic RNase cleavage assay (NIRCA). NIRCA suggested that the mutation site was near either amino acid position 58 of mature TTR or the 3' end of exon 3. Direct DNA sequencing showed both a normal GAG (Glu) and a variant AAG (Lys) codon at amino acid position 89 of mature TTR, which has not been previously reported. The site of this mutation is near the 3' end of exon 3, consistent with the result of NIRCA. This mutation was also confirmed by polymerase chain reaction-induced mutation restriction analysis (PCR-IMRA). PMID- 10842707 TI - Tabulation of transthyretin (TTR) variants as of 1/1/2000. PMID- 10842708 TI - Hereditary cystatin C amyloid angiopathy. PMID- 10842709 TI - 125I-labeled ApoE binds competitively to beta(1-40) fibrils with pathological chaperone proteins. AB - Radiolabeled Apolipoprotein E (Apo E) was used in a competitive binding filtration assay to amyloid fibrils preformed from beta(1-40) peptide as a probe of the binding sites for proteins either found in senile plaques in Alzheimer's Disease brain or reported to be associated with the soluble peptide. Apo E, Apo J, Apo A-I, Apo B, laminin, complement components C3 and C4, and alpha 1 antichymotrypsin all displayed sub-micromolar apparent affinities for the Apo E binding site on fibrils. Transthyretin, alpha 2-macroglobulin, amyloid P protein, heparan sulfate proteoglycan, complement component C1q, chondroitin sulfate A, and GM1 ganglioside were much less effective. The epsilon 2, epsilon 3, and epsilon 4 isoforms of Apo E showed different affinities for fibrils and lipidation of these lipoproteins made little difference. Other fibrillar beta peptides also bound Apo E, with A beta 40-A beta 42 > A beta(12-28); A beta(25 35) = 0. A series of soluble beta-peptides and fragments failed to effect Apo E binding. Thus, both conformational and quaternary structural features are important in high affinity binding of Apo E to A beta 40 fibrils. Different amyloid plaque-associated molecules apparently associate with alternative primary and secondary structural features on fibrils. PMID- 10842710 TI - beta-Amyloid(25-35) inhibits the activity of inositol(1,4,5)-trisphosphate-5 phosphatase. AB - beta-amyloid (A beta) peptides are known to disrupt calcium homeostasis in cells. In the present study, the effects of A beta(25-35) upon the activity of soluble Ins(1,4,5)P3-5-phosphatase have been investigated. A beta(25-35) inhibited, and dithiothreitol (DTT) increased the activity of soluble rat cerebellar Ins(1,4,5)P3-5-phosphatase. The change in activity was not accompanied by an obvious change in the sensitivity of the Ins(1,4,5)P3-5-phosphatase to inhibition by glucose-6-phosphate or phytic acid. A beta(35-25) also inhibited soluble Ins(1,4,5)P3-5-phosphatase activity, but at a lower potency than A beta(25-35). It is concluded that A beta(25-35) affects the metabolism of Ins(1,4,5)P3 although the potency is not sufficiently high to contribute to any significant extent to the effects of this peptide upon calcium homeostasis. PMID- 10842711 TI - Fibrillar amyloid-beta production, accumulation, and recycling in transgenic mice pancreatic acinar cells and macrophages. AB - Amyloid-beta (A beta) production, accumulation, and recycling were examined by light and electron microscopy in the pancreas of transgenic mice (from 45 days to 22 months of age) that express the gene for the carboxy-terminal fragment of the human amyloid-beta protein precursor. Ultrastructural immunocytochemistry revealed four types of cells accumulating fibrillar A beta 1-40 in cytoplasmic vacuoles: acinar pancreatic cells, macrophages infiltrating stroma, epithelial cells of pancreatic ducts, and blood monocytes/macrophages in the lumen of pancreatic vessels. The ultrastructure of amyloid deposits suggests that each of these four types of cells produces fibrillar A beta. Three basic types of amyloid deposits were distinguished: primary vacuoles in different stages of amyloid aggregation and fibrillization, secondary vacuoles that are the product of fusion of primary vacuoles, and phagosome-like vacuoles with morphologically intact fibrillar amyloid and residues of ingested cells. Amyloid production in acinar pancreatic cells starts in mice younger than 45 days, progresses in 2- to 7-month old mice, and plateaus in the second year of life. In macrophages, amyloid appears in 60-day-old mice, and the increase in the number of macrophages and the amount of amyloid in their cytoplasm correlates with age. PMID- 10842712 TI - Serum amyloid A in Alzheimer's disease brain is predominantly localized to myelin sheaths and axonal membrane. AB - Immunohistochemical localization of the injury specific apolipoprotein, acute phase serum amyloid A (A-apoSAA), was compared in brains of patients with neuropathologically confirmed Alzheimer's disease (AD), multiple sclerosis (MS), Parkinson's disease (PD); Pick's disease (Pick's), dementia with Lewy bodies (DLB), coronary artery disease (CAD), and schizophrenia. Affected regions of both AD and MS brains showed intense staining for A-apoSAA in comparison to an unaffected region and non-AD/MS brains. The major site of A-apoSAA staining in both diseases was the myelin sheaths of axons in layers V and VI of affected cortex. A-apoSAA contains a cholesterol binding site near its amino terminus and is likely to have a high affinity for cholesterol-rich myelin. These findings, along with our recent evidence that A-apoSAA can inhibit lipid synthesis in vascular smooth muscle cells suggest that A-apoSAA plays a role in the neuronal loss and white matter damage occurring in AD and MS. PMID- 10842713 TI - Serum amyloid A is present in the capillaries and microinfarcts of hypertensive monkey brain: an immunohistochemical study. AB - Serum amyloid A (SAA) is a major inducible acute phase protein characterized as a transient injury specific constituent of high density lipoprotein. We investigated whether the acute phase SAA (A-apoSAA), as a marker of inflammation, is present in the brain of monkeys with surgically induced hypertension of 39 months duration. Sections from brains of normotensive monkeys (systolic blood pressure < 124 mmHg) and hypertensive monkeys (systolic blood pressure > 185 mmHg) were processed for immunohistochemistry with a rabbit polyclonal antiserum to human A-apoSAA. We found that A-apoSAA was present in hypertensive but not in normotensive brain sections. Staining was localized to capillary endothelial cells and occasionally to the entire vessel wall of the prefrontal cortex. Staining was also observed in the capillaries and in medium size vessels of the corona radiata, the head of the caudate and, to a smaller extent, in the putamen. Additionally, the A-apoSAA was present in cells forming a circular configuration within microinfarcts. These findings suggest that high blood pressure in the brain can result in either local production of A-apoSAA in the capillaries and within microinfarcts or uptake of A-apoSAA from the blood PMID- 10842714 TI - Analysis of serum amyloid A1 exon 4 polymorphism in Japanese population. AB - Nucleotide variation in the triplet codon coding for amino acid position 72 of human serum amyloid A1 (SAA1), which was suggested by amino acid sequence analysis, was analyzed here in order to identify the genomic sequences coding SAA1.2 and to characterize further the SAA1 allele frequency in a Japanese population. The SAA1 exon 4 was amplified by PCR and treated with Nco I. Sequencing of PCR products from genomic DNA of individuals who were heterozygous for the Nco I site revealed GGT (Gly) and GAT (Asp) at the position 72. The allele having 72Asp showed the exon 3 polymorphism coding 52Ala and 57Val. This allele should thus be identified as SAA1.2. Alleles with 72Gly were either 52Val and 57Ala(SAA1.1) or 52Ala and 57Ala (SAA1.3) or 52Ala and 57Val (SAA1.5). The frequency of SAA1 alleles in the 321 Japanese subjects was 0.310, 0.012, 0.347 and 0.330 for each SAA allel of 1.1, 1.2, 1.3 and 1.5, respectively. The presence of the SAA.4 allele was not evaluated. PMID- 10842715 TI - Transthyretin isoleucine-122 mutation in African and American blacks. AB - The gene frequency of the transthyretin (TTR) mutation (Val122Ile) was studied in African and African-American populations. The African populations analyzed included the Zulu and Xhosa of South Africa, and Yorubas from the city of Ibadan, Nigeria. The African-American population included patients at the Veterans Affairs (VA) Medical Center, Indianapolis, and newborns from a local hospital in Indianapolis. The Val122Ile TTR mutation was identified in 1 of 55 Zulu, 0 of 34 Xhosa, 0 of 9 Nigerian subjects, 5 of 51 Veteran patients, and 3 of 103 newborns. Assuming the 2.91% prevalence in newborns to be the norm, there is a significant increased prevalence in the VA patient population. PMID- 10842716 TI - Kaposi's sarcoma-associated herpesvirus gene sequences are detectable at low copy number in primary amyloidosis. AB - Primary amyloidosis (AL), like multiple myeloma (MM), results from a clonal proliferation of plasma cells. Recent detection of Kaposi's sarcoma-associated herpesvirus (KSHV) gene sequences in MM patients, although controversial, suggested that KSHV may also be present in AL. In the present study, we assayed for KSHV gene sequences in patients with primary AL independently in 2 laboratories. Nested polymerase chain reaction (PCR) was performed on DNA isolated from 21 bone marrow (BM) core biopsy samples to amplify orf26 and orf72, 2 regions of the KSHV genome. Eighteen of 21 (86%) BM core biopsy samples were KSHV PCR positive. BM aspirates from 16 of these 21 AL patients were cultured for 4-6 weeks to generate long term bone marrow stromal cells (LT-BMSCs), and 13 of 16 (81%) LT-BMSCs were also KSHV PCR positive. Results in all but 1 sample were consistent in the 2 laboratories. Sequencing of the PCR products in the 2 laboratories confirmed 94-98% and 95-98% homology to the published orf 26 and orf 72 KSHV gene sequences respectively, with interpatient base pair differences. Despite the presence of KSHV gene sequences, only 4/18 (22%) KSHV PCR positive patients demonstrated KSHV lytic antibodies by immunoblot assay. A sensitive assay performed on the BCBL-1 cell line confirmed the presence of KSHV at a very low copy number in AL. PCR using patient specific light chain gene primers also amplified DNA isolated from 2 AL BM core biopsies and 3 AL LT-BMSCs which were KSHV PCR positive, suggesting the presence of clonotypic cells. Our results therefore demonstrate KSHV gene sequences albeit at a very low copy number in the majority of BM core biopsies and LT-BMSCs from AL patients, and serological responses in only a minority of cases. Ongoing studies to identify viral transcripts and gene products will determine the biological relevance of KSHV in AL disease pathogenesis. PMID- 10842717 TI - Japanese monozygotic twins with familial amyloidotic polyneuropathy (FAP) (ATTR Val30Met). AB - Twenty-nine-year-old twin brothers having the amyloidogenic transthyretin (ATTR) Val30Met gene developed the clinical symptoms of familial amyloidotic polyneuropathy (FAP) in 1995. The twins had the same educational background and lived in the same district. FAP manifestations were similar in both cases, although electromyographic examinations revealed sensorimotor polyneuropathy in No. 1 and sensory polyneuropathy in No. 2. DNA analysis revealed that they were monozygotic twins. In addition to environmental factors, genetic factors may play an important role in determining the onset of FAP. PMID- 10842719 TI - Update on variant Creutzfeldt-Jakob disease. PMID- 10842718 TI - Heart failure caused by a novel amyloidogenic mutation of the transthyretin gene: ATTR Ala45Ser. AB - Cardiac failure in transthyretin (TTR) amyloidosis patients has been shown to be caused by different mutations in the TTR gene. In the present case, a 73-year-old man from Northern Sweden was evaluated for heart failure. Amyloid deposits were found in subcutaneous fat and in intestinal biopsies. The presence of a variant form of TTR was detected in the plasma by electrospray ionisation mass spectrometry (ESI-MS). The mutation was located by single-strand conformation polymorphism (SSCP) analysis of the TTR gene where a band shift was seen in exon 2. Direct sequencing of exon 2 revealed a single base-pair substitution (G1724T). This transversion results in an amino acid substitution at codon 45, alanine to serine (ATTR Ala45Ser). Mass spectrometry analysis excluded that the variant is a polymorphism, since no similar shift in molecular weight has been present in more than 200 control samples. Congo red and immunostaining of duodenum biopsy specimens confirmed the presence of systemic ATTR amyloidosis, and clinical examination, including echocardiography, found evidence of a restrictive cardiomyopathy. He had 10 years previously been operated for a bilateral carpal tunnel syndrome, but otherwise no symptoms were present that could be attributed to his systemic amyloidosis. No axonal polyneuropathy was noted at nerve conduction studies. This novel mutation is the second amyloidogenic TTR mutation found in the Swedish population. PMID- 10842720 TI - New structural information and update on liver transplantation in transthyretin associated amyloidosis. Report from the 4th International Symposium on Familial Amyloidotic Polyneuropathy and Other Transthyretin Related Disorders & the 3rd International Workshop on Liver Transplantation in Familial Amyloid Polyneuropathy, Umea Sweden, June 1999. PMID- 10842721 TI - Sexual orientation and the sleep-wake cycle: a preliminary investigation. AB - The sleep-wake cycle as a function of sexual orientation was investigated. Male and female, heterosexual and homosexual subjects completed a sexual-orientation questionnaire and four sets of recording sheets of activities for a 16-day period. Mean sleep duration was calculated from the record sheets using waking-up and going-to-sleep times. It was predicted that homosexual males and females would awake earlier, go to sleep later, and thus have shorter sleep duration compared to heterosexuals. This was supported by the results. Although a sexual orientation by sex interaction just failed to reach significance, additional analyses of the results indicated that differences in sleep duration between heterosexuals and homosexuals were markedly related to sex. The possibility that these results reflect dimorphic features of the human supra-chiasmatic nucleus (SCN) is discussed. PMID- 10842722 TI - Sexual motives, gender, and sexual behavior. AB - The roles of gender and the sexual motives of Love, Pleasure, Conformity, Recognition, Dominance, and Submission in numerous usual and unusual sexual behaviors were investigated. In a survey of 191 college undergraduates it was found that Love, Pleasure, Conformity, and Recognition motives, often in interaction with gender, were all important predictors of sexual behavior. Gender was the best predictor of initiating usual sexual behavior, whereas the Love motive was the best predictor of actually engaging in usual sexual behavior. Pleasure and Recognition in interaction with gender were the best predictors of engaging in unusual sexual behavior. None of the sexual motives predicted initiating unusual sexual behavior. Findings suggest that a variety of sexual motives may underlie sexual behavior. PMID- 10842723 TI - Familial aspects of male homosexuality. AB - Research has generally supported the existence of familial-genetic factors for male sexual orientation, but has not shed much light on the specific nature of those influences. Gay men with gay brothers provide the opportunity to examine several hypotheses. Sixty-six men, representing 37 gay male sibling pairs, completed questionnaires assessing behavior on various measures including childhood and adult gender nonconformity, timing of awareness of homosexual feelings, self-acceptance, and the quality of family relationships. Consistent with prior findings using twins, gay brothers were similar in their degree of childhood gender non-conformity, suggesting that this variable may distinguish etiologically (e.g., genetically) heterogeneous subtypes. The large majority of gay men with brothers knew about their own homosexual feelings before they learned about their brothers' homosexual feelings, suggesting that discovery of brothers' homosexuality is not an important cause of male homosexuality. PMID- 10842724 TI - Personality characteristics of male clients of female commercial sex workers in Australia. AB - The personality characteristics of male clients of female sex workers and their motivations for seeking the services of sex workers were examined. It was hypothesized that clients of sex workers would differ from nonclients in that they would adopt less feminine sex roles, exhibit lower social-sexual effectiveness, and show higher levels of sensation-seeking behavior. Sixty-six clients of sex workers and 60 nonclients volunteered to complete a short questionnaire to assess demographic characteristics, sex role, social-sexual effectiveness, and sensation-seeking behavior. Clients and nonclients did not differ on demographic variables such as age, education, marital status, or occupation. However, clients were significantly less feminine in sex-role orientation, scored lower in social-sexual effectiveness, and scored higher on sensation seeking. Closer analysis of the client group revealed the existence of two distinct subgroups. The first was characterized by low social-sexual effectiveness and appeared motivated to visit sex workers because of an interpersonal need for intimacy. The second was characterized by high sensation seeking and appeared motivated to visit sex workers because of a need for novelty and variety in sexual encounters. PMID- 10842725 TI - Patterns of union formation among urban minority youth in the United States. AB - Since 1990, several large surveys of sexual behavior have been conducted. In addition to collecting general information on sexual histories, such as number of partners in the previous year and whether subjects ever used condoms, these studies collected information on sexual behavior with specific partners, or "partnerships." The data are useful both for testing of substantive hypotheses about the determinants of behavior as well as for disease transition modeling. The objective of this paper is to use partnership histories to describe the union formation patterns of low-income youth living in Detroit. Data from the partnership histories will be used to illustrate the types of statistics that can be generated from these histories. Data will be presented on the number and types of unions (married/cohabiting, "knew well," "casual"), the frequency and duration of these unions, the types of intercourse reported in each type of union, the patterns of mixing by age and ethnic group in each type of union, concurrency in unions, and condom use in unions. PMID- 10842726 TI - Bioinformatics education--a UK perspective. PMID- 10842727 TI - The complexities of genome analysis, the Retroid agent perspective. AB - MOTIVATION: The sequences of Retroid agents from a wide diversity of organisms constitute the largest set of complete genomes currently available for the study of genomic architecture and the transfer of information within and between organisms. These agents are ubiquitous in Eukaryotes, comprising 50-90% of the genomic information in some cases. RESULTS: Analyses conducted for over a decade illustrate that Retroid agents are engaged in a wide spectrum of molecular evolutionary events. A description of these complexities is presented as a three parameter conceptual framework that considers type, size, and mechanism of events that contribute to the evolution of genes, genomes, and organisms. The results of new data mining studies further illustrate the complexity of the network of relationships among and between Retroid agents and other organisms. A hidden Markov model construction strategy is presented that generates a multiple alignment more similar to those refined by human experts. CONTACT: mars@parvati.msu.montana. edu PMID- 10842728 TI - Self-organization and competition in the immune response to cancer invasion: a phase-orientated computational model of oncogenesis. AB - MOTIVATION: Recent studies indicate that fractal dimensions can uncover aspects of cellular dynamics prior to pathological manifestation. In this respect we are interested in building a computational model of oncogenesis able to generate patterns with the same fractal dimension spectrum as the in vivo tumor. RESULTS: A new theoretical model incorporating a systemic view of oncogenesis in a computational model was proposed. The tumor growth is viewed as competition for resources between the two self-organizing subsystems: the neoplastic and the immune. Numerical simulations revealed that tumor escape can be uncovered in some earlier stage of the immune-system-tumor interaction using multifractal measures. The described computational model is able to simulate also the case of immune, surgical, chemical and radiotherapeutical treatment, as well as their effects. AVAILABILITY: T The software used is available on request from the authors. CONTACT: Sorinel Oprisan, University of New Orleans, Department of Psychology, Ne w Orleans, LA 70148, USA. soprisan@uno.edu PMID- 10842729 TI - Darwin v. 2.0: an interpreted computer language for the biosciences. AB - MOTIVATION: We announce the availability of the second release of Darwin v. 2.0, an interpreted computer language especially tailored to researchers in the biosciences. The system is a general tool applicable to a wide range of problems. RESULTS: This second release improves Darwin version 1.6 in several ways: it now contains (1) a larger set of libraries touching most of the classical problems from computational biology (pairwise alignment, all versus all alignments, tree construction, multiple sequence alignment), (2) an expanded set of general purpose algorithms (search algorithms for discrete problems, matrix decomposition routines, complex/long integer arithmetic operations), (3) an improved language with a cleaner syntax, (4) better on-line help, and (5) a number of fixes to user reported bugs. AVAILABILITY: Darwin is made available for most operating systems free of char ge from the Computational Biochemistry Research Group (CBRG), reachable at http://chrg.inf.ethz.ch. CONTACT: darwin@inf.ethz.ch PMID- 10842730 TI - Sequence search algorithm assessment and testing toolkit (SAT). AB - MOTIVATION: The Sequence Search Algorithm Assessment and Testing Toolkit (SAT) aims to be a complete package for the comparison of different protein homology search algorithms. The structural classification of proteins can provide us with a clear criterion for judgment in homology detection. There have been several assessments based on structural sequences with classifications but a good deal of similar work is now being repeated with locally developed procedures and programs. The SAT will provide developers with a complete package which will save time and produce more comparable performance assessments for search algorithms. The package is complete in the sense that it provides a non-redundant large sequence resource database, a well-characterized query database of proteins domains, all the parsers and some previous results from PSI-BLAST and a hidden markov model algorithm. RESULTS: An analysis on two different data sets was carried out using the SAT package. It compared the performance of a full protein sequence database (RSDB100) with a non-redundant representative sequence database derived from it (RSDB50). The performance measurement indicated that the full database is sub-optimal for a homology search. This result justifies the use of much smaller and faster RSDB50 than RSDB100 for the SAT. AVAILABILITY: A web site is up. The whole packa ge is accessible via www and ftp. ftp://ftp.ebi.ac.uk/pub/contrib/jong/SAT http://cyrah.ebi.ac.uk:1111/Proj/Bio/SAT http://www.mrc-lmb.cam.ac.uk/genomes/SAT In the package, some previous assessment results produced by the package can also be found for reference. CONTACT: jong@ebi.ac.uk PMID- 10842731 TI - ProtEST: protein multiple sequence alignments from expressed sequence tags. AB - MOTIVATION: An automatic sequence searching method (ProtEST) is described which constructs multiple protein sequence alignments from protein sequences and translated expressed sequence tags (ESTs). ProtEST is more effective than a simple TBLASTN search of the query against the EST database, as the sequences are automatically clustered, assembled, made non-redundant, checked for sequence errors, translated into protein and then aligned and displayed. RESULTS: A ProtEST search found a non-redundant, translated, error- and length-corrected EST sequence for > 58% of sequences when single sequences from 1407 Pfam-A seed alignments were used as the probe. The average family size of the resulting alignments of translated EST sequences contained > 10 sequences. In a cross validated test of protein secondary structure prediction, alignments from the new procedure led to an improvement of 3.4% average Q3 prediction accuracy over single sequences. AVAILABILITY: The ProtEST method is available as an Internet World Wide Web service http://barton.ebi.ac.uk/servers/protest.html+ ++ The Wise2 package for protein and genomic comparisons and the ProtESTWise script can be found at http://www.sanger.ac.uk/Software/Wise2 CONTACT: geoff@ebi.ac.uk PMID- 10842732 TI - Fast assignment of protein structures to sequences using the intermediate sequence library PDB-ISL. AB - MOTIVATION: For large-scale structural assignment to sequences, as in computational structural genomics, a fast yet sensitive sequence search procedure is essential. A new approach using intermediate sequences was tested as a shortcut to iterative multiple sequence search methods such as PSI-BLAST. RESULTS: A library containing potential intermediate sequences for proteins of known structure (PDB-ISL) was constructed. The sequences in the library were collected from a large sequence database using the sequences of the domains of proteins of known structure as the query sequences and the program PSI-BLAST. Sequences of proteins of unknown structure can be matched to distantly related proteins of known structure by using pairwise sequence comparison methods to find homologues in PDB-ISL. Searches of PDB-ISL were calibrated, and the number of correct matches found at a given error rate was the same as that found by PSI BLAST. The advantage of this library is that it uses pairwise sequence comparison methods, such as FASTA or BLAST2, and can, therefore, be searched easily and, in many cases, much more quickly than an iterative multiple sequence comparison method. The procedure is roughly 20 times faster than PSI-BLAST for small genomes and several hundred times for large genomes. AVAILABILITY: Sequences can be submitted to the PDB-ISL servers at http://stash.mrc-lmb.cam.ac.uk/PDB_ISL/ or http://cyrah.ebi.ac.uk:1111/Serv/PDB_ISL/ and can be downloaded from ftp://ftp.ebi.ac.uk/pub/contrib/jong/PDB_+ ++ISL/ CONTACT: sat@mrc-lmb.cam.ac.uk and jong@ebi.ac.uk PMID- 10842733 TI - SAWTED: structure assignment with text description--enhanced detection of remote homologues with automated SWISS-PROT annotation comparisons. AB - MOTIVATION: Sequence database search methods often identify putative sub threshold hits of known function or structure for a given query sequence. It is widespread practice to filter these hits by hand using knowledge of function and other factors; to the expert, some hits may appear more sensible than others. SAWTED (Structure Assignment With Text Description) is an automated solution to this post-filtering problem which will be applicable to large scale genome assignments. RESULTS: A standard document comparison algorithm is applied to text descriptions extracted from SWISS-PROT annotations. The added value of SAWTED in combination with PSI-BLAST has been shown with a benchmark of difficult remote homologues taken from the SCOP structure database. AVAILABILITY: A WAWTED PSI BLAST Web server is available to perform sensitive searches against the protein structure database (http://www.bmm.icnet.uk/servers/sawted). CONTACT: R.MacCallum@icrf.icnet.uk PMID- 10842734 TI - TOPAL 2.0: improved detection of mosaic sequences within multiple alignments. AB - MOTIVATION: The Dss statistic was proposed by McGuire et al. (Mol. Biol. Evol., 14, 1125-1131, 1997) for scanning data sets for the presence of recombination, an important step in some phylogenetic analyses. The statistic, however, could not distinguish well between among-site rate variation and recombination, and had no statistical test for significant values. This paper addresses these shortfalls. RESULTS: A modification to the Dss statistic is proposed which accounts for rate variation to a large extent. A statistical test, based on parametric bootstrapping, is also suggested. AVAILABILITY: The TOPAL package (version 2) may be accessed from http:/ /www.bioss.sari.ac.uk/frank/Genetics and by anonymous ftp from typ://ftp.bioss.sari.ac.uk in the directory pub/phylogeny/topal. CONTACT: frank@bioss.sari.ac.uk PMID- 10842735 TI - TEXshade: shading and labeling of multiple sequence alignments using LATEX2 epsilon. AB - MOTIVATION: Typesetting, shading and labeling of nucleotide and peptide alignments using standard word processing or graphics software is time consuming. Available automatic sequence shading programs usually do not allow manual application of additional shadings or labels. Hence, a flexible alignment shading package was designed for both calculated and manual shading, using the macro language of the scientific typesetting software LATEX2 epsilon. RESULTS: TEXshade is the first TEX-based alignment shading software featuring, in addition to standard identity and similarity shading, special modes for the display of functional aspects such as charge, hydropathy or solvent accessibility. A plenitude of commands for manual shading, graphical labels, re-arrangements of the sequence order, numbering, legends etc. is implemented. Further, TEXshade allows the inclusion and display of secondary structure predictions in the DSSP-, STRIDE- and PHD-format. AVAILABILITY: From http://homepages.uni tuebingen.de/beitz/tse.h tml (macro package and on-line documentation) CONTACT: eric.beitz@uni-tuebingen.de PMID- 10842736 TI - Single column discrepancy and dynamic max-mini optimizations for quickly finding the most parsimonious evolutionary trees. AB - MOTIVATION: In the maximum parsimony (MP) method, the tree requiring the minimum number of changes (discrepancy) to explain the given set of DNA or amino acid sequences is chosen to represent their evolutionary relationships. To find the MP tree, the branch-and-bound algorithm is normally used. For a partial phylogenetic tree (one that has a subset of the organisms) the traditional algorithm assigns a cost equal to the discrepancy of the partial phylogenetic-tree. We propose a single column discrepancy heuristic which increases this cost by predicting a minimum additional discrepancy needed to attach the sequences yet to be added to the partial phylogenetic-tree. A dynamic Max-mini order of sequence addition is also proposed to quickly terminate branch-and-bound search paths that are guaranteed to lead to suboptimal solutions. RESULTS: We studied the running time of 47 problems generated from 17 data sets. The use of single column discrepancy heuristic speeded up the computation to 2.4-fold for static and 18.2-fold for dynamic search order. The improvement appeared to increase exponentially with the number of sequences. The proposed strategies are also likely to be useful in speeding up the MP tree search using heuristic searches that are based on branch and-bound-like algorithms. CONTACT: s.kumar@asu.edu PMID- 10842737 TI - Modeling splice sites with Bayes networks. AB - MOTIVATION: The main goal in this paper is to develop accurate probabilistic models for important functional regions in DNA sequences (e.g. splice junctions that signal the beginning and end of transcription in human DNA). These methods can subsequently be utilized to improve the performance of gene-finding systems. The models built here attempt to model long-distance dependencies between non adjacent bases. RESULTS: An efficient modeling method is described which models biological data more accurately than a first-order Markov model without increasing the number of parameters. Intuitively, a small number of parameters helps a learning system to avoid overfitting. Several experiments with the model are presented, which show a small improvement in the average accuracy as compared with a simple Markov model. These experiments suggest that single long distance dependencies do not help the recognition problem, thus confirming several previous studies which have used more heuristic modeling techniques. AVAILABILITY: This software is available for downloaded and as a web resource at http://www.ai.uic.edu/software CONTACT: kasif@eecs.uic.edu PMID- 10842738 TI - STAR/mmCIF: an ontology for macromolecular structure. AB - MOTIVATION: Crystallographers were motivated 10 years ago to develop a simple and consistent data representation for the exchange and archiving of data associated with the crystallographic experiment and the final structure. As this process evolved (and the data grew at near exponential rates) came the recognition that this representation should also facilitate the automated management of the data and, with the aid of additional software for verification and validation, provide improved consistency and accuracy and hence improved scientific inquiry. This realization led to a new Dictionary Definition Language (DDL) and an extensive dictionary based on this DDL for describing macromolecular structure. In broad terms this could be considered an ontology. An important feature in the development of the ontology was the endorsement and ongoing maintenance and support of the International Union of Crystallography (IUCr). While the description of macromolecular structure and the x-ray crystallographic experiment used to derive it represent explicit data, the ontology is extensible and applicable to other less well-characterized data domains. RESULTS: Details of the DDL, the dictionaries that have been developed, and software for reading and using this ontology are presented. AVAILABILITY: Extensive documentation, software tools and the DDL and dictionaries are available from http://ndbserver.rutgers.edu/mmcif and associated mirror sites. CONTACT: Bourne: bourne@sdsc.edu and Westbrook:jwest@rcsp.rutgers.edu PMID- 10842739 TI - PNAD-CSS: a workbench for constructing a protein name abbreviation dictionary. AB - MOTIVATION: Since their initial development, integration and construction of databases for molecular-level data have progressed. Though biological molecules are related to each other and form a complex system, the information is stored in the vast archives of the literature or in diverse databases. There is no unified naming convention for biological object, and biological terms may be ambiguous or polysemic. This makes the integration and interaction of databases difficult. In order to eliminate these problems, machine-readable natural language resources appear to be quite promising. We have developed a workbench for protein name abbreviation dictionary (PNAD) building. RESULTS: We have developed PNAD Construction Support System (PNAD-CSS), which offers various convenient facilities to decrease the construction costs of a protein name abbreviation dictionary of which entries are collected from abstracts in biomedical papers. The system allows the users to concentrate on higher level interpretation by removing some troublesome tasks, e.g. management of abstracts, extracting protein names and their abbreviations, and so on. To extract a pair of protein names and abbreviations, we have developed a hybrid system composed of the PROPER System and the PNAD System. The PNAD System can extract the pairs from parenthetical paraphrases involved in protein names, the PROPER System identified these paris, with 98.95% precision, 95.56% recall and 97.58% complete precision. AVAILABILITY: PROPER System is freely available from http://www.hgc.inc.u tokyo.ac.jp/service/tooldoc /KeX/intro.html. The other software are also available on request. Contact the authors. CONTACT: mikio@ims.u-tokyo.ac.jp PMID- 10842740 TI - The human transcript database: a catalogue of full length cDNA inserts. AB - SUMMARY: Full length cDNA sequences are an important resource for the research community but are currently intermingled with other sequences. We have identified the human full length insert cDNA sequences in GenBank and placed them in a single location, the Human Transcript Database. AVAILIBILITY: The Human Transcript Database is available at http://www.hgsc.bcm.tms.edu/HTDB/. CONTACT: John Bouck: jbouck@bcm.tmc.edu PMID- 10842741 TI - Dotlet: diagonal plots in a web browser. AB - Dotlet is a program for comparing sequences by the diagonal plot method. It is designed to be platform-independent and to run in a Web browser, thus enabling the majority of researchers to use it. AVAILABILITY: The applet can be tested at http://www.isrec.isb-sib.ch/java/dotlet/ Dotlet.html, and the source code is available upon request. CONTACT: Thomas. Junier Marco.Pagni @isrec.unil.ch SUPPLEMENTARY: The full documentation about d o t l e t is available from the above URL. PMID- 10842742 TI - SISEQ: manipulation of multiple sequence and large database files for common platforms. AB - A multiple sequence file converter for common platforms, SISEQ,is described, which performs extraction of DNA sequences that correspond to CDS or RNA field of a large database file as well as subsequent multi-sequence conversions for phylogenetic or molecular biological analysis. Command-line interface as well as a GUI and a script-driven operation mode are provided. AVAILABILITY: The program is freely available to academic users in the form of Macintosh FAT binary, DOS executable, or UNIX source code at http://www.molbiol.saitama-u.ac.jp/naoki/ Software.html. CONTACT: naokisat@molbiol.saitama-u.ac.jp PMID- 10842743 TI - GOLD--graphical overview of linkage disequilibrium. AB - SUMMARY: We describe a software package that provides a graphical summary of linkage disequilibrium in human genetic data. It allows for the analysis of family data and is well suited to the analysis of dense genetic maps. AVAILABILITY: http://www.well.ox.ac.uk/asthma/GOLD CONTACT: goncalo@well.ox.ac.uk PMID- 10842744 TI - TAMBIS: transparent access to multiple bioinformatics information sources. AB - TAMBIS (Transparent Access to Multiple Bioinformatics Information Sources) is an application that allows biologists to ask rich and complex questions over a range of bioinformatics resources. It is based on a model of the knowledge of the concepts and their relationships in molecular biology and bioinformatics. AVAILABILITY: TAMBIS is available as an applet from http://img.cs.man.ac.uk/tambis SUPPLEMENTARY: A full manual, tutorial and videos can be found at http://img.cs.man.ac.uk/tambis. CONTACT: tambis@cs.man.ac.uk PMID- 10842745 TI - Cellular thiols and redox-regulated signal transduction. AB - In contrast to the conventional notion that reactive oxygen is mostly a trigger for oxidative damage of biological structures, now we know that low physiologically relevant concentrations of ROS can regulate a variety of key molecular mechanisms that may be linked with important cell functions (Fig. 4). Redox-based regulation of gene expression has emerged as a fundamental regulatory mechanism in cell biology. Several proteins, with apparent redox-sensing activity, have been described. Electron flow through side-chain functional CH2-SH groups of conserved cysteinyl residues in these proteins account for the redox sensing properties. Protein thiol groups with high thiol-disulfide oxidation potentials are likely to be redox-sensitive. The ubiquitous endogenous thiols thioredoxin and glutathione are of central importance in redox signaling. Signals are transduced from the cell surface to the nucleus through phosphorylation and dephosphorylation chain reactions of cellular proteins at tyrosine and serine/threonine. Protein phosphorylation, one of the most fundamental mediators of cell signaling, is redox-sensitive. DNA-binding proteins are involved in the regulation of cellular processes such as replication, recombination, viral integration and transcription. Several studies show that the interaction of certain transcription regulatory proteins with their respective cognate DNA sites is also redox-regulated. Changes in the concentration of Ca2+i control a wide variety of cellular functions, including transcription and gene expression; Ca(2+)-driven protein phosphorylation and proteolytic processing of proteins are two major intracellular events that are implicated in signal transduction from the cell surface to the nucleus. Intracellular calcium homeostasis is regulated by the redox state of cellular thiols, and it is evident that cell calcium may play a critical role in the activation of the redox-sensitive transcription factor NF-kappa B. Among the several thiol agents tested for their efficacy in modulating cellular redox status, N-acetyl-L-cysteine and alpha-lipoic acid hold most promise for human use. A strong therapeutic potential of strategies that would modulate the cellular thioredoxin system has been also evident. PMID- 10842746 TI - Integration of antagonistic signals in the regulation of nitrogen assimilation in Escherichia coli. PMID- 10842747 TI - Regulation of nuclear import and export. PMID- 10842748 TI - Regulation of glutathione synthesis. PMID- 10842749 TI - Superoxide dismutase, oxidative stress, and cell metabolism. PMID- 10842750 TI - Regulation of axonal neurofilament phosphorylation. PMID- 10842751 TI - Thiol-based antioxidants. AB - The thiol redox status of intracellular and extracellular compartments is critical in the determination of protein structure, regulation of enzyme activity, and control of transcription factor activity and binding. Thiol antioxidants act through a variety of mechanisms, including (1) as components of the general thiol/disulfide redox buffer, (2) as metal chelators, (3) as radical quenchers, (4) as substrates for specific redox reactions (GSH), and (5) as specific reductants of individual protein disulfate bonds (thioredoxin). The composition and redox status of the available thiols in a given compartment is highly variable and must play a part in determining the metabolic activity of each compartment. It is generally beneficial to increase the availability of specific antioxidants under conditions of oxidant stress. Cells have devised a number of mechanisms to promote increased intracellular levels of thiols such as GSH and thioredoxin in response to a wide variety of stresses. Exogenous thiols have been used successfully to increase cell and tissue thiol levels in cell cultures, in animal models, and in humans. Increased levels of GSH and other thiols have been associated with increased tolerance to oxidant stresses in all of these systems and in some cases, with disease prevention or treatment in humans. A wide variety of thiol-related compounds have been used for these purposes. These include thiols such as GSH and its derivatives, cysteine and NAC, dithiols such as lipoic acid, which is reduced to the thiol form intracellularly, and "prothiol" compounds such as OTC, which are enzymatically converted to free thiols within the cell. In choosing a thiol for a specific function (e.g., protection of lung from oxidant exposure or protection of organs from ischemia reperfusion injury), the global effects must also be considered. For example, large increases in free thiols in the circulation are associated with toxic effects. These effects may be the result of thiyl radical-mediated reactions but could also be due to destabilizing effects of increases in thiol/disulfide ratios in the plasma, which normally is in a more oxidized state than intracellular compartments. Changes in the thiol redox gradient across cells could also adversely affect any transport or cell signaling processes, which are dependent on formation and rupture of disulfide linkages in membrane proteins. Therapeutic thiol administration has been shown to have great potential, and its efficacy should be increased by selecting compounds and methods of delivery that will minimize perturbations in the thiol status of regions external to the targeted areas. PMID- 10842752 TI - Regulation and role of heme oxygenase in oxidative injury. AB - The HO-1 isoenzyme is an early stress response gene regulated by many forms of oxidative stress. The HO-2 isoenzyme is predominantly a constitutive enzyme, which may serve to sequester heme as well as degrade it. All HO enzyme activity results in the degradation of heme and the production of antioxidant bile pigments, which would favor an antioxidant role for the enzyme. In fact, in oxidative stress in vitro, HO-1 is protective (91-94) but within a narrow threshold of overexpression (93,94) in some models, since iron released in the HO reaction may obviate any cytoprotective effect (Fig. 3). So far, HO-2 appears to be beneficial in oxygen toxicity in vivo, but the consequences of HO-2 overexpression have not yet been tested. It will be important to better define the role of each HO isoenzyme in oxidative stress so as to determine whether enhancing these complex systems could alleviate some of the cellular changes seen as a result of oxidative injury. Furthermore, prior to considering therapeutic maneuvers to enhance HO, a complete understanding of the physiologic consequences of HO-1 induction and associated reactions, in each particular setting, will be crucial. PMID- 10842753 TI - Antioxidant regulation of genes encoding enzymes that detoxify xenobiotics and carcinogens. AB - Antioxidants are substances that delay or prevent the oxidation of cellular oxidizable substrates. The various antioxidants exert their effect by scavenging superoxide or by activating a battery of detoxifying/defensive proteins. In this chapter, we have focused on the mechanisms by which antioxidants induce gene expression. Many xenobiotics (e.g., beta-naphthoflavone) activate genes similar to those activated by antioxidants. The promoters of these genes contain a common cis-element, termed the antioxidant response element (ARE), which contains two TRE (TPA response element) or TRE-like elements followed by GC box. Mutational studies have identified GTGAC***GC as the core of the ARE sequence. Many transcription factors, including Nrf, Jun, Fos, Fra, Maf, YABP, ARE-BP1, Ah (aromatic hydrocarbon) receptor, and estrogen receptor bind to the ARE from the various genes. Among these factors, Nrf-Jun heterodimers positively regulate ARE mediated expression and induction of genes in response to antioxidants and xenobiotics. This Nrf-Jun heterodimerization and binding to the ARE requires unknown cytosolic factors. The mechanism of signal transduction from antioxidants and xenobiotics includes several steps: (1) Antioxidants and xenobiotics undergo metabolism to generate superoxide and related reactive species, leading to the generation of a signal to activate expression of detoxifying/defensive genes. (2) The generation of superoxide and related reactive species is followed by activation of yet to be identified cytosolic factors by unknown mechanism(s). (3). Activated cytosolic factors catalyze modification of Nrf and/or Jun proteins, which bind to the ARE in promoters of the various detoxifying/defensive genes. (4) The transcription of genes encoding detoxifying/defensive proteins is increased. The unknown cytosolic factors are significant molecules because they represent the oxidative sensors within the cells. Identification of the cytosolic factors will be of considerable importance in the field of antioxidants and gene regulation research. Future studies will also be required to completely understand the molecular mechanism of signal transduction from antioxidants and xenobiotics to Nrf-Jun. In addition to the Nrf-Jun pathway, mammalian cells also contain other pathways that activate gene expression in response to oxidative stress. These include NF-KB-, HIF-1-, Mac-1-, and SRF-mediated pathways. It is expected that collectively these pathways increase transcription of more than four dozen genes to protect cells against oxidative stress. PMID- 10842754 TI - Modulation of vascular cell activation, function, and apoptosis: role of antioxidants and nuclear factor-kappa B. AB - The activity of NF-kappa B is critically involved in the inflammatory activation of endothelial cells and their adhesiveness and also appears to regulate apoptosis in SMC by coordinating antiapoptotic programs. The activity of NF-kappa B has been revealed within human atheromas or following angioplasty but not in undiseased arteries. Hence, the inhibition of NF-kappa B mobilization by antioxidative or anti-inflammatory agents or by adenoviral I kappa B alpha overexpression, as reviewed herein, may act in concert to suppress endothelial activation and to induce SMC apoptosis. This synergistic concept may be a vasoprotective approach to prevent atherogenesis and restenosis by attenuating inflammatory reactions and SMC proliferation in nascent and progressing atherosclerotic lesions, as well as in developing neointima formations following angioplasty. PMID- 10842755 TI - Calcineurin: from structure to function. PMID- 10842756 TI - Microembolus detection by transcranial Doppler ultrasound. PMID- 10842757 TI - Dichhaptic task: methodological factors and brain organization. AB - Significant effects of shape, sequence of shapes, distractors, order of report and practise were demonstrated in a free recall dichhaptic task with nonsense shapes in adults. A left hand superiority (in accordance with theoretical expectations) was observed only in the second reports of the first half of the test. No sex differences were found. There was a preference to report first the right hand, and the opposite preference was observed in a dichhaptic letter task. It is probable that all these methodological factors may have affected previous dichhaptic studies. The association with other lateral and cognitive parameters is also analysed. PMID- 10842758 TI - Childhood diabetic neuropathy: a clinical and electrophysiological study. AB - Clinical diabetic neuropathy in childhood is rare, but electrophysiological involvement of the peripheral nerve is more frequent. We assessed clinically and electrophysiologically the peripheral nervous system of 69 children and adolescents suffering from diabetes mellitus (DM). The mean age of the patients was 12.8 years and the mean age at onset of DM was 6.8 years with a mean disease duration of 6.3 years. Seven patients (10%) had clinical neuropathy of which ankle jerk reflex abolition was the most frequent sign. Twenty patients (29%) had a neurophysiological neuropathy prevalently affecting the lower limbs. Peripheral neuropathy was correlated with patient age, older age at onset, duration of DM, height and poor glycaemic control. PMID- 10842759 TI - Gastric electromechanical dysfunction in Parkinson's disease. AB - This study was designed to evaluate gastric myoelectrical and mechanical activities in idiopathic Parkinson's disease (IPD) patients. Twenty patients with IPD (14 male and 6 female, mean age 42 +/- 9 years) were studied. Patients were divided into two groups: group A--early stage of disease (no. = 6) and group B- advanced IPD (no. = 14). Electrogastrography (EGG) was performed in fasting and postprandial conditions (Synectics system). The cross-sectional area of the gastric antrum was measured by sonography (Hitachi EUB-240). The antral area in fasting conditions was 2.1 +/- 0.4 and 4.2 +/- 1.2 cm2 and gastric emptying was 75 +/- 5 and 125 +/- 12 min in groups A and B respectively. EGG showed dysrhythmias (range 1-6 cycles per minute) in about 75% of both groups of IPD patients without increase in signal amplitude after a meal. Our results suggest that gastric motility is particularly impaired in patients with advanced IPD. It may be caused by the primary degenerative process in the autonomic nervous system of the gut. PMID- 10842761 TI - What is psychoanalysis? PMID- 10842760 TI - MRI--diagnostic and follow-up tool for microprolactinomas. AB - Magnetic resonance imaging is a method of choice for the diagnosis of microprolactinomas. Using a Phillips Gyroscan NT 15 (1.5 T), 42 patients whose clinical history and serum prolactin levels were compatible with the diagnosis of prolactinomas were examined. Four patients (3 male and 1 female) with macroadenoma were investigated and excluded, while microprolactinoma was found in the other 38 (32 female and 6 male) patients. Imaging features were similar in all microprolactinomas--hypointensity up to 10 mm in size in T1 weighted images, only sometimes, in the case of a cystic or a hemorrhagic component, also registered in T2 weighted images. Deviation of the pituitary stalk was present in 19 of the 38 patients (16 with microprolactinoma, 3 without any detectable lesion). Convex upper limit of the gland was demonstrated in 17 subjects (16 microprolactinomas, 1 without detectable lesion). Pituitary adenoma is not the only cause of raised serum prolactin level. This can be a consequence of medication, spinal cord tumor, compression of the pituitary stalk, hypothyroidism or a lesion in dermatomes T3 to T5 (mammary gland level). Results from imaging must always be assessed in the light of the patient's clinical history and biochemical parameters. PMID- 10842762 TI - Interactional parapraxes: a window onto our representations of self and others. PMID- 10842763 TI - Consequences of a male therapist disclosing the birth of his child. PMID- 10842764 TI - Cross-cultural dimensions of sadomasochism in the psychoanalytic situation. PMID- 10842765 TI - The analyst's relation to the good. PMID- 10842766 TI - The development of discernment in psychotherapy and Quaker worship. PMID- 10842767 TI - Rorschach interaction patterns, alexithymia, and closeness to parents in psychotic and psychosomatic patients. PMID- 10842768 TI - Oedipus as normative? Freud's complex, Hook's query, Malinowski's trobrianders, Stoller's anomalies. PMID- 10842769 TI - Shame dynamics in the psychotherapy of the patient with PTSD: a viewpoint. PMID- 10842770 TI - The good girl grows up: gender and self-esteem. PMID- 10842771 TI - Eroticism and the conventional. PMID- 10842772 TI - Cutaneous manifestations in anorexia nervosa. AB - Anorexia nervosa is an eating disorder among adolescent girls and young women which, though common, often goes undetected and untreated. Anorexia nervosa is a response for young people with psychological conflicts who try to win love by having a body corresponding to the present-day image, symbolising strength, beauty, attraction, power and success. Anorexia nervosa involves inadequate calorie intake leading to marked cachexia with metabolic and endocrinological disturbances. We investigated dermatological changes in 21 young female anorectics aged 19-24 in an attempt to find dermatological markers which mirror the dynamics of the disease and thus obtain helpful signs for early diagnosis with its important bearing on the outcome. Extensive histories were taken and whole-body examinations performed. Seven sex- and age-matched persons served as a control group. The most common dermatological findings were xerosis (71%, controls 29%), cheilitis (76%), bodily hypertrichosis (62%), alopecia (24%), dry scalp hair (48%), acral coldness (38%), acrocyanosis (33%), periungual erythema (48%), gingival changes (37%), nail changes (29%) and calluses on dorsum of hand due to self-induced vomiting (67%). Our study documented for the first time that a body mass index of < or = 16 (kg/m2) can be considered a critical value at which skin changes are more frequent. There are remarkable similarities between cutaneous manifestations in anorexia nervosa and in HIV infection. Patients with anorexia nervosa develop early stereotype skin changes which are cardinal diagnostic symptoms and pointers to the diagnosis of eating disorders. During training at the Department of Child and Adolescent Psychiatry in Solothurn one of us (C. H.) was once more able to observe most of the above-described cutaneous and mucocutaneous changes in anorexic adolescents. This paper is intended to stimulate further basic research on this topic. We hope our study will facilitate early diagnosis of anorexia nervosa by the family physician and enable him or her to institute immediate treatment for the eating disorder and thereby improve the prognosis. PMID- 10842773 TI - Characteristics of Caucasian type 2 diabetic patients during ketoacidosis and at follow-up. AB - OBJECTIVE: To analyse among adult Caucasian patients hospitalised for diabetic ketoacidosis the relative frequency of patients with type 2 diabetes and their characteristics. RESEARCH DESIGN: A retrospective review of adult patients presenting with diabetic ketoacidosis was conducted between 1993 and 1996. Patients with typical type 1 diabetes were classified according to age of onset < 35 years, insulin-dependence and BMI < 25 kg/m2; patients who did not meet these criteria were further classified on the basis of a basal and stimulated C-peptide and the detection of antibodies to glutamic acid decarboxylase (GADA) and to islet cells (ICA). RESULTS: 43 patients presenting with an episode of diabetic ketoacidosis were divided into two groups, A and B. Group A consisted of 19 patients, 17 patients classified as patients with typical type 1 diabetes and 2 patients with diabetes post-pancreatectomy. The other patients were the subjects of our study (group B; n = 20, 4 lost to follow-up). 13 patients (65% of group B = B I) were diagnosed with type 1 diabetes (median basal C-peptide: 0.15 nmol/l) and 7 patients (35% of group B = B II) with type 2 diabetes (median basal C peptide of 1 nmol/l). Higher body mass index (BMI), shorter duration of diabetes, smaller anion gap and worse glycaemic control were found to be significantly different between groups B I and II on admission (p < 0.05). After a median follow-up of 18 months, patients in group B II had a better metabolic control than those in B I and 5 of the 7 patients were treated without insulin. CONCLUSION: Diabetic ketoacidosis is more common than previously thought in patients with type 2 diabetes, occurring in 16% of all cases. Distinctive features at presentation are the degree of acidosis, the duration of diabetes, BMI and HbA1c. However, the basal plasma C-peptide value remains the best discriminating factor. PMID- 10842774 TI - [A further case of analbuminemia]. AB - At routine follow-up a 23-year-old female presented a high erythrocyte sedimentation rate and handicapping lipodystrophy of the lower limbs. Protein electrophoresis showed absence of an albumin peak and the diagnosis of analbuminaemia was therefore proposed. Investigation of family members disclosed that one brother out of a total of four siblings also had analbuminaemia. The hereditary pathway often appears to be autosomalrecessive. Subsequent review of the literature revealed only 28 other observed cases, although today electrophoresis has become a worldwide routine examination. Surprisingly, the almost complete absence of so important a protein as albumin does not trigger disease. The body is able to compensate for the lack of albumins with other proteins. The absence of abumin is typically associated with hyperlipidaemia and lipodystrophy in the female sex. PMID- 10842775 TI - [Zenker's diverticulum]. AB - The first case of posterior pharyngooesophageal diverticulum was published in 1764 by Ludlow. Zenker's name has been attributed to the diverticulum since his description of a series of patients in 1878. The aetiology and pathogenesis of Zenker's diverticulum are not well understood. Research has mainly focused on the role of the upper oesophageal sphincter, but numerous manometric studies have produced controversial results. Also, the influence of gastrooesophageal reflux on the upper oesophageal sphincter and the development of a diverticulum is unclear. Patients with a Zenker's diverticulum typically present with a long history of slowly progressive dysphagia for solid consistencies and regurgitation of undigested food. Weight loss and nocturnal attacks of coughing may bother the patient. The diagnosis of a diverticulum needs to be confirmed by radiologic examination. The only definite therapy is surgery. The classical extramucosal cricopharyngeal myotomy by transcervical approach, with or without removal of the diverticular sac, is increasingly giving way to transmucosal myotomy through a transoral endoscopic approach. Compared to the transcervical approach the endoscopic technique avoids the risk of injuring the recurrent laryngeal nerve, substantially lowers the number of pharyngeal fistulas and, in large series, showed an equivalent outcome as far as relief of symptoms is concerned. In the light of the literature and our own experience diverticulooesophagostomy with the Endo-GIA stapler by a transoral endoscopic approach has become the therapy of choice at the ENT-Department of the University Hospital of Zurich, Switzerland. In an operating time of only 10-15 minutes the stapler cuts the wall between the diverticular sac and the oesophagus, and in the same manoeuvre closes the mucosal wound edges with tiny staples. Oral feeding is possible from the first postoperative day. With the technique described this elderly population of patients obtains rapid and safe relief of symptoms. PMID- 10842776 TI - [Just a full bladder?]. PMID- 10842777 TI - Nutrition in cancer: an overview. AB - OBJECTIVES: To provide a review of weight loss, cachexia (both primary and secondary), and weight gain in cancer patients. DATA SOURCES: Research reports, review articles, textbooks, and personal communications. CONCLUSIONS: Alterations in nutritional status have the potential to affect mortality, morbidity, and quality of life outcomes. The detection and treatment of malnutrition is important to success of therapy. IMPLICATIONS FOR NURSING PRACTICE: Nursing interventions to help maintain optimal nutritional status in patients with cancer include careful assessment, identification of patients at risk, and management of problems before the initiation of therapy. PMID- 10842778 TI - Dietary factors and cancer risk. AB - OBJECTIVES: To discuss the relationship between dietary factors and cancer risk reduction, the status of prevention trials in progress, and current dietary recommendations. DATA SOURCES: Published research and review articles, medical books, and web sites. CONCLUSIONS: Evidence indicates that dietary factors can reduce the risk of developing cancer. The dietary recommendations for cancer risk reduction are consistent with those for prevention of other chronic diseases and are thus useful for the majority of the population. IMPLICATIONS FOR NURSING PRACTICE: In their role as educators, it is essential for nurses to provide accurate and timely information on diet and cancer risk reduction to their patients and the public and to encourage compliance with dietary recommendations. PMID- 10842779 TI - Nutritional screening and assessment. AB - OBJECTIVES: To review the components of nutritional screening, screening measures and tools, and the nurses' role in nutritional assessment. DATA SOURCES: Research studies, review articles, and book chapters. CONCLUSIONS: Timely and appropriate nutritional interventions require the adoption of routine initial nutritional screening, referral for comprehensive nutritional assessments as needed, and continued re-screening. IMPLICATIONS FOR NURSING PRACTICE: Oncology nurses should be prepared to contribute to a comprehensive nutritional assessment in their role as patient advocate and expert clinician. PMID- 10842780 TI - Symptoms and their impact on nutrition. AB - OBJECTIVES: To provide assessment guidelines and management strategies for symptoms affecting nutritional status in patients with cancer. DATA SOURCES: Published articles, book chapters, and research reports. CONCLUSIONS: Patients with cancer experience a variety of symptoms that affect their nutritional status. It is possible to reduce the impact of these symptoms on the nutritional status of the patient through effective symptom management strategies, nutritional assessments, and interventions. IMPLICATIONS FOR NURSING PRACTICE: The nutritional status of cancer patients can be supported by anticipating and managing disease- and treatment-related symptoms. Nurses contribute actively to the achievement of positive patient outcomes when they manage symptoms in a manner that enhances nutrition. PMID- 10842781 TI - Nutrition support in cancer. AB - OBJECTIVES: To describe nutrition intervention strategies frequently used to support cancer patients experiencing malnutrition. DATA SOURCES: Textbook chapters, database reports, and current related literature. CONCLUSIONS: Clinicians have many options for providing nutrition support interventions for the cancer patient at risk for malnutrition. The most appropriate strategy for a patient should be based on a careful assessment of contributing factors with input from a multidisciplinary team. IMPLICATIONS FOR NURSING PRACTICE: It is important for nurses to be knowledgeable about nutrition intervention options available to patients at various points along the cancer trajectory. Oncology nurses are in a key position to provide support to patients and families with regard to nutrition issues. Of paramount importance is their contribution to ongoing assessment of nutritional status and early and aggressive intervention to meet nutritional needs. PMID- 10842782 TI - Nutritional care of surgical oncology patients. AB - OBJECTIVES: To review nutritional management of surgical oncology patients, particularly patients with head and neck cancer and gastrointestinal malignancies. DATA SOURCES: Professional journal articles, published research findings, and book chapters related to the nutritional management of surgical oncology patients. CONCLUSIONS: Meeting the nutritional needs of oncology patients is challenging. A multidisciplinary approach to the nutritional management of the surgical oncology patient is essential to ensure comprehensive care and assistance in the recovery process. IMPLICATIONS FOR NURSING PRACTICE: Nurses need to have an understanding of the impact of both the disease process and surgical intervention on the nutritional needs of oncology patients to improve the quality of care and quality of life for these special populations. PMID- 10842783 TI - Nutritional issues in palliative care. AB - OBJECTIVES: To present current information on nutritional problems and management during the palliative care period. DATA SOURCES: Research and review articles from three computerized databases, the table of contents of an on-line nursing journal, and pursuit of pertinent references in articles reviewed. CONCLUSIONS: Controversies continue on the most appropriate management of intake in the palliative care period from both physical and ethical perspectives. IMPLICATIONS FOR NURSING PRACTICE: Until the literature is conclusive on the amount and type of suffering incurred by patients who are ceasing intake, nurses will need to stay current with the research literature and approach each case individually in determining appropriate care. PMID- 10842784 TI - Nutritional considerations in pediatric oncology. AB - OBJECTIVES: To summarize the characteristics of a nutrition care plan for pediatric patients, specifically pediatric screening, assessment, and treatment protocols. DATA SOURCES: Review articles, manuals, and textbooks. CONCLUSIONS: Nutritional support of pediatric patients with cancer differs from adults because of their unique nutritional needs, their wide age range (infants to adolescents), and the psychosocial aspects of treating the entire family. IMPLICATIONS FOR NURSING PRACTICE: To participate in the prevention and treatment of malnutrition in the pediatric patient with cancer, nurses require an understanding of nutrition care in this specialized population. PMID- 10842785 TI - Nutritional issues in patients with severe neutropenia. AB - OBJECTIVES: To provide information on the assessment and management of the nutritional needs of patients with severe neutropenia. DATA SOURCES: Textbook chapters, research articles, and review articles. CONCLUSIONS: Malnutrition is a common result of high-dose chemotherapy treatment. Clinical outcomes are directly related to careful assessment, early intervention, and ongoing evaluation and monitoring in the severely neutropenic population. IMPLICATIONS FOR NURSING PRACTICE: Early assessment, management, and psychological support are essential in meeting the needs of this patient population. PMID- 10842786 TI - Nutrition as a component of alternative therapy. AB - OBJECTIVES: To discuss common "alternative" nutritional strategies that may be used by patients as a primary or adjunctive means of cancer treatment. DATA SOURCES: Government reports, textbook chapters, published articles, and research reports. CONCLUSIONS: The use of diet and nutrition to prevent and treat disease is one of six fields of alternative medicine. A number of dietary regimens have been purported to be effective in the treatment of cancer. Most have been inadequately tested with regard to safety and efficacy. IMPLICATIONS FOR NURSING PRACTICE: Oncology nurses need to be knowledgeable of the use of alternative methods so that they may provide accurate information to patients considering these approaches. PMID- 10842787 TI - Rethinking sanitation: panacea or Pandora's box. AB - In the 19th century, sanitation solutions were designed and built on the premises that human excreta was a waste suitable only for disposal and that the environment was capable of assimilating the waste. The prevailing view last century was that vapors from smells caused disease, and the best way to deal with excreta was to convey it to rivers and streams where it could be diluted and cleansed. Times have changed, the premises are outdated, and current solutions contribute, either directly or indirectly, to many of the problems faced by society today: water pollution, scarcity of fresh water, food insecurity, destruction and loss of soil fertility, loss of biodiversity, depletion of the ozone layer, and global warming. A common denominator of all these problems is how society deals with its wastes, specifically how it deals with human excrement. We have to rethink past premises, design and build new systems, and contribute to the solving of society's most pressing problems. The panacea of the 19th century is turning out to be the pandora's box of the 21st century. PMID- 10842788 TI - Re-engineering the urban drainage system for resource recovery and protection of drinking water supplies. AB - The Harare metropolis in Zimbabwe, extending upstream from Manyame Dam in the Upper Manyame River Basin, consists of the City of Harare and its satellite towns: Chitungwiza, Norton, Epworth and Ruwa. The existing urban drainage system is typically a single-use-mixing system: water is used and discharged to "waste", excreta are flushed to sewers and eventually, after "treatment", the effluent is discharged to a drinking water supply source. Polluted urban storm water is evacuated as fast as possible. This system not only ignores the substantial value in "waste" materials, but it also exports problems to downstream communities and to vulnerable fresh-water sources. The question is how can the harare metropolis urban drainage system, which is complex and has evolved over time, be rearranged to achieve sustainability (i.e. water conservation, pollution prevention at source, protection of the vulnerable drinking water sources and recovery of valuable materials)? This paper reviews current concepts regarding the future development of the urban drainage system in line with the new vision of "Sustainable Cities of the Future". The Harare Metropolis in Zimbabwe is taken as a case, and philosophical options for re-engineering the drainage system are discussed. PMID- 10842789 TI - Some recent and ongoing initiatives for the development of strategies, models and tools in the field of environmental sanitation in economically less developed countries. PMID- 10842790 TI - Health impairments arising from drinking water resources contaminated with Vibrio cholerae. AB - The endemic and seasonal nature of cholera depends upon the survival of toxigenic Vibrio cholerae in various niches of the aquatic environment. To understand the transmission and ecology of V. cholerae, it is necessary to know which component in the aquatic ecosystem can harbor it and thus contribute to the endemic presence. Toxigenic V. cholerae is now recognized as an autochthonous member of the microflora in many aquatic environments based on its protracted survival and proliferation without losing the virulence determinants. This article summarizes knowledge about the ecology, survival strategies and elimination techniques of V. cholerae from natural waters with special reference to drinking water. PMID- 10842791 TI - Efficiency of wastewater treatment plants at removing Giardia sp. cysts in southern France. AB - A study was conducted to evaluate the removal efficiency for Giardia sp. cysts of a number of wastewater treatment plants in France. Of these, five were activated sludge systems, three were trickling filters and three were waste stabilisation pond systems. In addition, the effect upon cyst removal of disinfecting effluents prior to discharge into controlled waters by UV and chlorination was evaluated. Cysts were detected in raw wastewater at all 11 treatment plants in concentrations ranging from 130 to 41,270 cysts/litre. The removal of cysts by sewage treatment was found to range between 99.5 and 99.8% for activated sludge, 99.9 and 100% for waste stabilisation ponds, and for the trickling filter plants up to 98.3%. Despite the high removal efficiencies recorded in this study the range of cysts detected in final effluents discharged into controlled waters ranged from < 1 to 66 cysts/litre. This is undoubtedly an underestimate of actual occurrence and concentration in wastewater, given the limitations of the methods currently employed by investigators. Cysts were detected in final effluents which had been disinfected by both UV and chlorination (range 0.3-19 cysts/litre), however, it is not known whether such cysts were viable or non-viable. PMID- 10842792 TI - Health impairments arising from drinking water polluted with domestic sewage and excreta in China. AB - Raw water of poor quality still causes many drinking-water associated health problems all over China, largely because of poor sanitation, inadequate disposal of sewage and excreta. Eutrophication due to excess of total nitrogen and phosphorous in some sources for drinking-water has led to massive proliferation of cyanobacteria. The dominant species of cyanophyta can produce microcystins, a potent liver cancer promotor. As in previous studies, high incidence of liver cancer coincided with high microcystin concentration in the source water, especially in pond water. A frequent consequence of heavy pollution of source water is further the high incidence of infectious intestinal diseases, which are more than 10-100 times as frequent in China than in developed countries. PMID- 10842793 TI - Water, sanitation and diarrhoea. AB - Water, sanitation and health are inseparably linked. Diarrhoea and other water related diseases are the major causes of health problems in developing countries. Although the need for water and sanitation interventions for health promotion has been recognised, these are labeled as costly and are often neglected in the primary healthcare programmes. Lack of proper water and sanitation initiatives based on appropriate techniques, technologies, knowledge and/or implementation methods have hindered the expected achievements through water and sanitation interventions. Since water and sanitation initiatives include both availability of provisions and their effective use (which mean behavioral changes), they are technically and socially challenging. Disasters and emerging water quality problems, such as arsenic in groundwater, have further been complicating this situation. After reviewing relevant articles, several research issues are suggested in the context of developing country perspectives. PMID- 10842794 TI - Challenges for health and water resources in the Birim districts of eastern Ghana. AB - In the late 1970s and early '1980s, the Ghana Water and Sewerage Corporation (GWSC) with assistance from the government of Germany (KfW through IGIP) drilled 144 wells in the Birim North and South Districts of the Eastern Region of Ghana. This was part of an overall provision of water to rural communities which later came to be known as the 3000 Wells Project. The wells were supply-driven and were centrally-maintained, without community involvement in planning and operation. Earlier in 1970, an agricultural development project was also promoted in the two otherwise deprived districts with the aim of increasing output of produce and thereby improving incomes of rural farmers with emphasis on cocoa, rice and oilpalm. In 1992, under a United Nations' Development Programme Rural Water and Sanitation Project with the GWSC, another 141 wells were drilled and assistance provided to households to build 750 Ventilated Improved Pit (VIP) latrines coupled with hygiene education under a demand-driven community management approach. (About 60% of the budget was allocated to water, another 30% to sanitation while some 10% was meant for hygiene education interventions). All the wells were fitted with modified Indian Mk II pumps. These measures together were meant to improve the health of rural communities and make them contribute more positively to national development. The paper looks at some of the possible factors of the two water projects and the agricultural development project that could impact negatively on the water resources and health of people in these two districts. PMID- 10842795 TI - Drugs, diagnostic agents and disinfectants in wastewater and water--a review. AB - After administration pharmaceuticals are excreted by the patients into the aquatic environment via wastewater. Unused medications are sometimes disposed of in drains. The drugs may enter the aquatic environment and eventually reach drinking water, if they are not biodegraded or eliminated during sewage treatment. Additionally, antibiotics and disinfectants are assumed to disturb the wastewater treatment process and the microbial ecology in surface waters. Furthermore, resistant bacteria may be selected in the aeration tanks of sewage treatment plants by the antibiotic substances present. Since the 1980s, data on the occurrence of pharmaceuticals in natural surface waters and the effluents of sewage treatment plants have been reported. More recently, pharmaceuticals have been detected in ground and drinking water. However, only little is known about the risk imposed on humans by pharmaceuticals and their metabolites in surface and drinking water. An overview of input, occurrence, elimination (e.g. biodegradability) and possible effects of different pharmaceutical groups such as anti-tumour drugs, antibiotics and contrast media as well as AOX resulting from hospitals effluent input into sewage water and surface water is presented. PMID- 10842796 TI - Establishing mutual trust--a prerequisite to public partnership. AB - Establishing mutual trust is the key to establishing workable and sustainable partnerships between the public and service providers, in any part of the world. The experience of utilities in the United Kingdom and United States shows that trust is established through sharing information with the public and improving customer service. Understanding the needs of the public and the service agency are a prerequisite to establishing trust. There is no "magic formula" for the process, as it depends on the current level of knowledge, attitudes and actual practice (behavior) of both the public and the service agency. In Jordan, for example, there is awareness of environmental issues, but trust must be developed to generate public support for programs. PMID- 10842797 TI - Management of rural drinking water supplies and waste using the participatory hygiene and sanitation transformation (PHAST) initiative in Zimbabwe. AB - This paper focuses on the use of Participatory Hygiene and Sanitation Transformation (PHAST) and how the methodology can be taken to scale. It uses the Zimbabwe experience and highlights some of the benefits in the application of PHAST, conditions necessary for scaling up and possible constraints. The PHAST initiative started off as a pilot process seeking to promote improved hygiene behaviour and promotion of sanitation. Having successfully piloted PHAST, Zimbabwe has scaled up the use of the methodology at a country level. While impact studies have not yet been conducted, reviews of the effects of the process have indicated positive behaviour change in such areas as management of water, construction and use of latrines. The process has also led to a change of institutional approaches in planning for improved water and sanitation from supply driven projects to demand responsive approaches. Some lessons learnt have included the need for baseline surveys at the start of the use of PHAST, the difficulty in developing monitoring indicators and hence difficulty in measuring impacts. Conclusions being drawn using assessment studies are that the use of participatory approaches has led to improved hygiene behaviour with communities being able to link causes and effects. The use of participatory methods also necessitates a change in institutional approaches from supply driven approaches to demand responsiveness. Other lessons drawn were related to the creation of an enabling environment for the application of participatory processes. Such enabling environment includes capacity building, resource allocation, policy and institutional support. PMID- 10842798 TI - Protecting drinking water: water quality testing and PHAST in South Africa. AB - The paper presents an innovative field-based programme that uses a simple total coliform test and the approach of PHAST (Participatory Hygiene And Sanitation Transformation) to help communities exploring possible water quality problems and actions that can be taken to address them. The Mvula Trust, a South African water and environmental sanitation NGO, has developed the programme. It is currently being tested throughout South Africa. The paper provides two case studies on its implementation in the field, and suggests ways in which the initiative can be improved in the future. PMID- 10842799 TI - The development of a protocol to manage the potential of groundwater contamination from on-site sanitation. AB - The provision of water supply and sanitation services to the developing and water scarce areas of South Africa is one of the goals of the Reconstruction and Development Programme. One of the principles of the South African National Sanitation Policy stipulates that sanitation systems which have unacceptable impacts on the environment cannot be considered to be an adequate form of sanitation. The greatest contamination concern with respect to ventilated improved pit toilets is in relation to groundwater resources. To this end the National Sanitation Co-ordination Office (NaSCO) and the Directorate of Geohydrology of the Department of Water Affairs and Forestry have developed "A Protocol to Manage the Potential of Groundwater Contamination from On-Site Sanitation". The aim of the protocol is to ensure that reasonable measures are taken to guard against contamination of valuable groundwater resources by inappropriately located or designed sanitation systems. The protocol focuses on dry on-site sanitation systems and provides a brief background to the current understanding of groundwater contamination by on-site sanitation systems. It also presents a procedure to be followed by those involved in implementing new, or upgrading old systems--the procedure allows the field worker to assess whether groundwater resources may be at risk of pollution. The protocol is specifically aimed at engineering and technical staff who are not groundwater specialists. PMID- 10842800 TI - Regulation as a tool for improving the management of water supply and sanitation companies. AB - The establishment of a strong independent regulator appears to be the most successful means to remedy the endemic poor management of water supply and sanitation services. This paper discusses why this is so and illustrates the argument with reference to recent experience in Chile. PMID- 10842801 TI - Water management for development of water quality in the Ruhr River basin. AB - On the Ruhr, a small river running through hilly country and with a mean flow of 76 m3/s, 27 water works use the method of artificial groundwater recharge to produce 350 million m3 of drinking water annually. On the basis of a special act, the Ruhr River Association is responsible for water quality and water quantity management in the Ruhr basin. The present 94 municipal sewage treatment plants ensure that the raw water is sufficiently good to be turned into drinking water. In the Ruhr's lower reaches, where dry weather results in a 20% share of the entire water flow being treated wastewater, comparatively high concentration of substances of domestic or industrial origin are likely, including substances which municipal wastewater treatment measures cannot entirely remove. These substances include ammonium, coliform bacteria or pathogens, boron and organic trace substances. Although water treatment measures have greatly contributed to the considerable improvement of the Ruhr's water quality in the last few decades, it is desirable to continue to aim at a high standard of drinking water production technologies since the Ruhr is a surface water body influenced by anthropogenic factors. However, in the case of substances infiltrating into drinking water, legislation is required if a reduction of pollution appears to be necessary. PMID- 10842802 TI - New regulatory framework for wastewater quality control in Mexico. AB - This paper analyses the new regulatory framework for wastewater quality control developed in Mexico. It provides a description of the Mexican government strategies for water pollution control. The discussion focuses on the policy instruments used, their evolution and the main difficulties encountered in their implementation. The new regulatory framework is discussed highlighting some of the economic implications of the reforms. The paper concludes that proper institutional development and monitoring are essential factors for the success of any policy instrument for wastewater quality control. PMID- 10842803 TI - Pollution of water sources due to poor waste management--the case of Dar-es Salaam. AB - Pollution of water sources for the city of Dar-es-Salaam originates from haphazard disposal of solid wastes, discharge of untreated or inadequately treated wastewater to water sources, lack of standard sanitary facilities and poor hygienic practices. Contaminated water used for human consumption can lead to serious health problems e.g. cholera, typhoid, skin diseases, etc., which, in turn, leads to reduced working hours/manpower. This has a direct effect to production output which can lead to a deterioration of local community welfare. Having realised this as a problem, the Government of Tanzania stipulated, in its water policy of 1991, the need for protection of water sources. In achieving this goal, proper waste management was singled out to be of vital importance. Due to economic hardships, however, budget allocation by the central Government could not cover the costs needed for proper handling of waste. This left Tanzania with no alternative other than heavy reliance on donor and bilateral organisations for financial support of programmes. Nevertheless, these sources of funds proved to be unreliable for many different reasons. To deal with these problems, the Government currently emphasises involving local community and NGOs, the formation of stakeholder funds and organisations, and involvement of the private sector. Other efforts are intensification of education programmes to create more awareness to the local communities on the need for protection of water sources. Although at its infancy level, the system is showing some signs of improvement. PMID- 10842804 TI - Resource protection and resource management of drinking water-reservoirs in Thuringia--a prerequisite for high drinking-water quality. AB - In face of widespread pollution of surface waters, strategies must be developed for the use of surface waters which protect the high quality standards of drinking water, starting with the catchment area via the reservoir to the consumer. As a rule, priority is given to the avoidance of contaminants directly at their point of origin. Water protection is always cheaper than expensive water body restoration and water treatment. Complementary to the generally practised technical methods of raw water treatment with all their associated problems of energy input requirements, costs, and waste products, there is an increasing number of environmentally sound treatment technologies which use ecological principles as a basis to support the self-cleaning properties of flowing and dammed waters. PMID- 10842805 TI - Using the example of Istanbul to outline general aspects of protecting reservoirs, rivers and lakes used for drinking water abstraction. AB - The six main drinking water reservoirs of Istanbul are under the threat of pollution due to rapid population increase, unplanned urbanisation and insufficient infrastructure. In contrast to the present land use profile, the environmental evaluation of the catchment areas reveals that point sources of pollutants, especially of domestic origin, dominate over those from diffuse sources. The water quality studies also support these findings, emphasising that if no substantial precautions are taken, there will be no possibility of obtaining drinking water from them. In this paper, under the light of the present status of the reservoirs, possible and probable short- and long-term protective measures are outlined for reducing the impact of point sources. Immediate precautions mostly depend on reducing the pollution arising from the existing settlements. Long-term measures mainly emphasise the preparation of new land use plans taking into consideration the protection of unoccupied lands. Recommendations on protection and control of the reservoirs are stated. PMID- 10842806 TI - Wastewater discharge and water quality standards in Brazil--implications for the selection of wastewater treatment technologies. AB - The paper describes and comments the Brazilian standards for water quality and effluent discharge (CONAMA Directive No. 20, 1986). The emphasis of the paper is on the conventional parameters which characterise domestic wastewater (BOD, SS, NH4+, N, P and coliforms). The wastewater treatment systems for achieving compliance are analysed with two perspectives: requirements for the discharge standards and requirements for the receiving water standards. The main treatment technologies available are listed, together with their capability to remove the major pollutants included in the Brazilian legislation. The analysis for the receiving water standards are given for different dilution ratios (river/discharge flows): 1/10, 1/1, 10/1 and 100/1. The difficulty in complying with the receiving water standards for BOD, phosphate and coliforms is highlighted and the cost implications for the adoption of more sophisticated technologies in order to achieve compliance are also provided. Although the specific quantitative values cited in the paper are associated with the Brazilian legislation, it is believed that the major conceptual issues are applicable to a large number of urban areas in other countries. PMID- 10842807 TI - Monitoring of Cryptosporidium and Giardia in Czech drinking water sources. AB - In Czech raw water sources for drinking water supply, Cryptosporidium was found in numbers from 0 to 7400 per 100 liters and Giardia from 0 to 485 per 100 liters. The summer floods of 1997 probably brought the highest numbers of Cryptosporidium oocysts into one of the reservoirs sampled; since then these numbers decreased steadily. A relatively high number of Cryptosporidium oocysts was found in one sample of treated water. Repeated sampling demonstrated that this was a sporadic event. The reason for the presence of Cryptosporidium in a sample of treated drinking-water is unclear and requires further study. PMID- 10842808 TI - The suitability of sand abstraction systems for community-managed domestic water supplies. AB - This paper will discuss the system of abstracting water from the underlying water bearing sediment of surface dry river beds. The technology is well established in Zimbabwe and is a valuable method of obtaining water for human consumption as well as for both livestock and irrigation use. Within Zimbabwe the system is erroneously but commonly, referred to as "sand abstraction". Apart from traditional methods there are several systems of sand abstraction which have been developed. Each method is dependant on equipment which can be installed into river sands which retain water. Systems range from large mechanized schemes to small scale, hand-operated pump and well screen units. The author, through his work and experience in this area has come to believe that small scale units which can be operated and sustained by local communities are the best option for reliable sources of clean water. In large rivers the sediment frequently retains a perennial supply of water and during a dry-season can provide many thousands of kilolitres of water. The river sands act as a huge, slow sand-filtration system and invariably yield water which is neither contaminated by poor sanitation nor by unpalatable mineral salts. Tests undertaken on water samples drawn from river sands have indicated that contamination occurs only after abstraction from containers. Water utilised from such sources is thus particularly suitable for both primary and secondary purposes. Where such systems can be established on low river banks, not too distant from the river edge, simple sand abstraction units provide an excellent, low cost method of abstracting water. Such systems are particularly appreciated by women who prefer small, low technology installations which they can operate themselves and can maintain simply and effectively. PMID- 10842809 TI - Groundwater protection policies and practices in the United Kingdom. AB - This paper summarises the main objectives for groundwater protection and describes the system adopted in the UK. Groundwater is essential as a source of drinking-water, even in countries with high rainfall; it is used for irrigation and industrial purposes; and it often comprises the base flow of rivers under dry conditions. Water quality is threatened by industry, landfill leachate, agricultural contamination and mine drainage. Quantity is threatened by over abstraction. The UK uses a risk-based concept of vulnerability to pollution and over-abstraction. A groundwater protection policy has been devised providing guidance on those activities which require control in groundwater protection zones, reflecting the vulnerability of the aquifer. The zones make use of the travel time of contaminants to the water abstraction point. All major activities such as water abstraction, waste disposal, and spreading of agricultural materials may thus be assessed in terms of their risk to the groundwater, and suitable precautions may be taken. Groundwater, once polluted, is an asset which is difficult and expensive to replace. Vulnerability assessment is a useful tool to assist in its protection. PMID- 10842810 TI - Conflict of technologies for water and sanitation in developing countries. AB - Borehole water supplies, in basement rock aquifers in the West Africa Sub-region, face potential pollution hazards as a result of their close location within the same geological environments as indiscriminately sited latrines, rubbish dumps, farms and animal watering points in the communities. The heterogeneous nature of the overburden and fractures in the bedrock constitute relatively fast flow paths for surface water contaminated mainly by bacteria and nitrates which enhance the pollution of the groundwater and boreholes. To improve the drinking water quality, some measures have been taken to minimize the hazards. Further studies are required to understand better the nature and scale of the problem and to avoid the apparent conflict of technologies. It is necessary to incorporate improvements in sanitation into rural water supply projects, if the otherwise good drinking water source should not be lost to society's wastes. PMID- 10842811 TI - Pit latrine effluent infiltration into groundwater: the Epworth case study. AB - Water can be a vehicle for the transmission of communicable diseases. Technologies have been developed to protect groundwater from external surface contamination. However, there is growing concern about the likelihood of pit latrine effluent infiltrating into groundwater reservoirs for well water supply systems. Investigations on seasonal variations and extent of pit latrine effluent infiltration into soil and groundwater have been carried out in Zimbabwe. Preliminary results show that groundwater flows in the direction of surface runoff, and that there is no lateral soil pollution above the groundwater surface. Pit latrine contents leach downwards and down slopes for distances that vary per season and soil type. The results also demonstrate the contribution of refuse pits and water collection methods to groundwater pollution. PMID- 10842812 TI - A comparison of the extent and impacts of sewage contamination on urban groundwater in developed and developing countries. AB - In much of the world urban groundwater is an important resource for domestic and industrial use. In many developing countries, groundwater taken directly (untreated) from individual springs and wells is the only option available to communities where comprehensive, reliable reticulated supply systems are absent. A common feature of urban groundwater in both developing and developed countries is contamination by sewage. Current and recent research is presented that shows sewer leakage impacts groundwater in developed countries whilst on-site sanitation contaminates groundwater in developing countries. In the latter case, the competing demands of sanitation and groundwater protection must be addressed. Limitations on the usefulness of accepted standard sewage indicator species in groundwater are also highlighted. As sewage contamination of groundwater is usually addressed only if an actual health risk is posed, it is vital both to developed and developing countries to understand the movement of actual pathogens in groundwater in the context of groundwater management. Further research is required on microbial survival and health risks posed by sewage contamination. PMID- 10842813 TI - Sanitary survey of the drinking water supply of Kombinati suburb-Tirana, Albania. AB - Microbiological pollution of drinking water is a major health problem in the suburbs of the Albanian capital. Intermittent supply and contamination, resulting in several gastrointestinal manifestations, are the main concerns for the population and health workers. The risk of outbreaks of water-borne diseases is high. Pollution originates from contamination of drinking water with domestic sewage. This research investigated the drinking water cycle from its natural source to the consumer, analysing samples and verifying pollution levels in the microbiological and chemical setting. The most important pollution sources were found in the distribution network, due to cross-contamination with sewers and illegal connections. The second pollution source was found around the extraction wells. This is related to abusive constructions within the sanitary zone around the wells and maybe the highly sewage-contaminated river water which feeds the aquifer. PMID- 10842814 TI - Nitrate pollution study in the aquifer of Dakar (Senegal). AB - Dakar is a peninsula inhabited by a population of about 2 million people in 1996. In some dug wells and piezometers, the nitrate content (NO3.) in the groundwater is above the World Health Organization (WHO) limit of 50 mg/l. In the unconfined part of the aquifer of the peninsula, all the samples from wells are contaminated by high nitrate contents which increased over time from 100 mg/l in 1987 to more than 250 mg/l in 1996. Only a limited area is affected by nitrate pollution in the confined layer. The results indicate anthropogenic pollution, a fact which indicates the increasing risk of pollution of drinking-water resources. Studies in the unsaturated zone and familiarity with the sanitation practices in the area indicate that the horizontal and vertical flux are linked mainly to defective septic tanks and direct organic waste elimination into the soil by more than 40% of the inhabitants. The correlation between tritium values (3H) and nitrate shows that the source of nitrate is recent. The relation of oxygen 18 (18O) to deuterium (2H) in water with high nitrate levels indicates that the concentrations of nitrate have been identified in evaporated points. PMID- 10842815 TI - Tools of groundwater protection below the city of Caracas, Venezuela. AB - Caracas city obtains drinking water from neighbouring catchment areas. Since these water resources are limited in quantity, groundwaters under the city are also considered for drinking water supply. Hydrochemical and isotope investigations show that the active recharge zone which may readily be contaminated reaches to about 50 m below floor. At greater depths the passive recharge zone extends to a maximum of 300 m and is by far less susceptible to groundwater pollution than the active recharge zone. The water balance indicates recharge to the Caracas aquifer of 2.1 m3/s from losses of the distribution and collector systems of waters as well as from subsurface lateral groundwater inflow into the Caracas valley. The active recharge zone of the aquifer beneath Caracas actually acts as an important microbiological reactor. It was proposed that exploitation of 20% of the total amount of groundwater re-charge from depths below 100 m would be sustainable and provide unpolluted water. Abstraction from the active recharge zone would require protection measures in the city area, which are not feasible. PMID- 10842816 TI - Improvements of wastewater treatment for groundwater protection in the Haertsfeld. AB - Wastewater treatment in rural water protection zones is crucial for sustainable source water protection. Specific problems arise where wastewater from decentral treatment plants infiltrates towards vulnerable aquifers. Within the water protection zone of the Egau Waterworks sited on the Swabian highlands with a karstic aquifer, a concept for a central sewer system and wastewater plant went into operation in 1993 in order to solve the conflict between drinking water demands and wastewater treatment. Since then, water quality of the Buchbrunnen karst spring which is used for drinking water supply for about 400,000 inhabitants has shown improvements. The paper describes the development of typical parameters including groundwater flow time and corresponding concentration levels. The concept implemented here may serve as a model for a successful water protection initiative. PMID- 10842817 TI - Wastewater reuse with groundwater safeguard. AB - In Morocco, reuse of raw wastewater for irrigation is practised around the big cities without taking into account the sanitary and environmental concerns. All national institutions involved in wastewater problems are convinced of the need for wastewater treatment with extensive systems before it can be applied for agricultural reuse. Our experimental work on wastewater treatment using lagooning, macrophytic plants, reed beds, infiltration percolation or over-land flow showed that of all these extensive systems lead to an effluent classified into category B according to WHO guidelines. However, the amount of nitrogen largely exceeded the crops' requirements and could cause serious problems for groundwater. Using the DRASTIC method, a vulnerability map was established for groundwater in the plain surrounding the city of Marrakech. The estimates of nitrogen amounts from wastewater that can reach groundwater after crop uptakes showed that only the low vulnerability zone could be safely irrigated with the treated wastewater. The second zone of moderate vulnerability covering 50% of the area in the plain could receive the treated wastewater, but a careful program of irrigation is required to avoid nitrate contamination of groundwater. The third zone at a high vulnerability level should be protected, without any further irrigation using wastewater, even after extensive treatment. This new approach integrating the results of the preliminary vulnerability study of groundwater zones could constitute a helpful tool to improve management of regional projects of wastewater reclamation and reuse. PMID- 10842818 TI - National Research Council report on potable reuse. AB - In 1998, the United States National Research Council prepared a report assessing the viability of potable reuse. The report concludes that planned indirect potable reuse is a viable application of reclaimed water but should be considered for implementation only after other alternative measures, such as development of new water sources, nonpotable reuse, and water conservation, have been evaluated and determined not to be economically or technically feasible. Although health effects research has not indicated that there have been any adverse health effects resulting from drinking highly treated reclaimed water, those data are sparse. There are uncertainties associated with assessing the potential health risks of drinking reclaimed water. The report acknowledges that several issues remain unresolved regarding reclaimed water treatment and reliability, microbial and chemical constituents of concern, and water quality and health effects monitoring. Research needs include methods to detect emerging pathogens, identification and quantification of trace organic compounds, development of suitable in vivo toxicological methods, and evaluation of the effectiveness and reliability of multiple treatment barriers. The report also concludes that direct potable reuse is not a viable option to consider at this time. PMID- 10842819 TI - Efficiency and sustainability of soil-aquifer treatment for indirect potable reuse of reclaimed water. AB - An increasing number of municipalities are considering the indirect reuse of treated wastewater (recycled water) by groundwater recharge as a feasible option to augment potable water supplies. This planned approach offers several advantages compared to the conventional way of discharging effluents into surface waters, including the additional treatment afforded as the water percolates and co-mingles with groundwater (soil aquifer treatment). While groundwater recharge has been used in the United States (U.S.) for several decades and has been the subject of a number of studies, limitations in methodology and testing have prevented many within the scientific and technical community from being able to fully address a number of complex public health questions related to organic chemicals, nitrogen and microorganisms. Ongoing research being conducted in Arizona and California is directed at reducing the uncertainties about the efficiency and sustainability of soil aquifer treatment for indirect potable reuse of recycled water. PMID- 10842820 TI - Reuse of wastewater in aboriginal communities in Western Australia. AB - Western Australia is a very arid area and freshwater is a precious resource. The wise and efficient use of this resource is essential for the establishment of communities in this region. Wastewater reuse in Aboriginal communities is a very recent development. Initially leach drains disposed of wastewater. Due to their failure most of the major communities now have the effluent from the septic tanks collected by a small diameter reticulated sewerage system and directed to oxidation ponds for treatment. The overflow from the oxidation ponds is allowed to flow over land or to a creek without any specific use. This paper discusses available reuse options as well as the options specific to Aboriginal communities in Western Australia. It is intended to demonstrate that in arid regions reuse wastewater can act as a water conservation and pollution control measure. PMID- 10842821 TI - Health effect of wastewater reuse in agriculture. AB - The present study addresses the impact of wastewater reuse in agriculture on the transmission of protozoan and helminthic infections. For this purpose, an epidemiological study was carried out on two populations of children totalling 608 individuals for protozoan infections and 528 for helminthic infections. Each population comprised an exposed group living in the wastewater spreading area of Marrakech and a control group. Results showed that 72% of the exposed group had protozoan infections. This rate did not exceed 45% in the control group. The pathogenic protozoan infections observed were giardiasis and amoebiasis. Regarding helminthic infections, 73% of the exposed children were infected compared with 30% of the control group. The risk attributable to wastewater reuse in the transmission of pathogenic protozoan and helminthic infections was 41% and 43%, respectively. Children of the spreading area are therefore more exposed to detectable risks from parasitic helminths and protozoa than the control children. PMID- 10842822 TI - Health risk in agricultural villages practicing wastewater irrigation in central Mexico: perspectives for protection. AB - 9,435 individuals participated in a cross-sectional survey in the irrigation districts of the Mezquital Valley (central Mexico). Exposure groups were: 848 households irrigating with untreated wastewater, 544 households irrigating with the effluent from a series of interconnected reservoirs, and 928 households farming with natural rainfall. The unit of analysis was the individual, and the health outcomes included diarrhoeal diseases and Ascaris lumbricoides infection. Water quality was assessed using faecal coliforms (FC) and nematode eggs, as suggested by (WHO, 1989) for the safe use of wastewater in agriculture. Children from households irrigating with untreated wastewater (10(8) FC/100 mL and 135 nematode eggs/L) had a 33% higher risk of diarrhoeal diseases and a fivefold increase in risk of A. lumbricoides infection (OR = 5.71) compared to children from the control group, farming with rainfall. The risk of A. lumbricoides infection in older individuals was even higher (OR = 13.18). The final analysis showed that drinking unboiled water and cultivating vegetables crops were both associated with a higher risk of diarrheal diseases (OR = 1.45 and 2.00); individuals infected with A. lumbricoides infection came mostly from landless households with poorer dwellings and low standards of sanitation (OR = 2.20, 2.23, 1.72 and 1.43, respectively). These results are discussed in the context of health protection measures and policy recommendations. PMID- 10842823 TI - Carbon, nitrogen and heavy metal dynamics in long-term wastewater irrigated Mexican soils. AB - In column experiments of Mexican wastewater irrigated soils the effects of changing water quality (untreated = UT, primary = PT and tertiary treated = TT wastewater) on carbon, nitrogen and heavy metal dynamics were investigated. In the column effluents the nitrate concentrations varied between 141-683 mg l-1. The total amount of leached TOC decreased in the order UT > PT > TT. Outflow concentrations of Pb ranged from 31.6-166.5 micrograms l-1 and of Cu from 31.2 146.8 micrograms l-1. Irrigation water quality influenced the Pb but not the Cu efflux. Cu seemed to be co-transported with TOC by preferential flow whereas there was no correlation of Pb and TOC concentrations in the effluents. The possibility of Pb transport through preferential flow paths is discussed. PMID- 10842824 TI - Evaluation of groundwater pollution potential of sewage-irrigated vegetable growing areas of the eastern fringe of Calcutta city. AB - In recent years recycling in agriculture is a common method of disposal or utilisation of waste. However, recycling of wastes may cause contamination of groundwater by toxic elements like heavy metals, cationic and anionic contaminants and pathogens. Groundwater of shallow and deep tubewells was collected during 1991 to 1997 from raw sewage effluent irrigated garbage farming areas on the eastern fringe of Calcutta city. In general raw sewage effluents, sludges and sewage-irrigated soils contain very high amounts of cations, anions, organics and heavy metals. It is found that most of the groundwater contained undesirable pH, total dissolved solids, total hardness, calcium, magnesium, phenolic compounds, iron and manganese and the observed values or concentrations were much above the maximum desirable limits specified by World Health Organisation (WHO) and Bureau of Indian Standards (BIS) for use as drinking water. Groundwater of that area may be used for irrigation. Dispersion by leaching of the metals, cationic and anionic contaminants from irrigated soil and from settled bottom sludge in unlined sewage channels are the principal causes of groundwater contamination. Some management plans have been suggested to control further deterioration of groundwater quality. PMID- 10842825 TI - Sanitation without pollution. AB - The most effective way of protecting drinking water resources from domestic sewage is to use technologies that do not produce sewage. This paper gives an overview of emerging alternatives in the form of ecological sanitation systems for urban and peri-urban areas. A key feature of ecological sanitation is that it regards human excreta as a resource to be recycled rather than as waste to be disposed of. Examples given include ecological sanitation systems based on dehydration and decomposition from Mexico, El Salvador, Sweden, India and Vietnam. These systems need neither water for flushing, nor pipelines for transport, nor treatment plants and arrangements for the disposal of toxic sludge. Large scale application of ecological sanitation would lead to less environmental pollution, reduced water consumption, considerable savings on sewers and treatment plants and increased employment. In addition it would provide valuable resources for food production and wasteland development. PMID- 10842826 TI - Reuse--the ultimate sink? Urine-diverting toilets to protect groundwater quality and fertilise urban agriculture. AB - People are concerned about water and food scarcity and the threats that faecal pollution and malnutrition pose to their health. Ecological sanitation systems open up for new, constructive options in sanitation, not least in poor periurban areas. The purpose of developing a no-mix excreta disposal system is to save water, to reduce wastewater treatment problems, and to protect groundwater quality as well as to recirculate nutrients from urine. In this paper all these aspects will be dealt with comprehensively. PMID- 10842827 TI - Measurement of bacterial growth potential in a reclaimed water. AB - The objectives of this paper are to show a bio-assay method using direct total microbial count (DTMC) and to demonstrate the applicability of this method to reclaimed water for evaluating its potential of bacterial growth. We have applied our method to four types of reclaimed water, measuring the time course of DTMC and dissolved organic carbon in a bio-assay process. The experimental findings are summarised as follows: 1) By regulating organic carbon, bacteria growth could be controlled. 2) Reduction of the initial concentration of bacteria was effective to delay their growth, but this operation could not control their maximum level. This implies that the retention time of the storage and distribution system of reclaimed water should be considered in design and operation of the disinfection process. 3) Ozonation increased the growth potential. Carbon adsorption process reduced it, but the ozone plus carbon adsorption process could not improve the biological stability. 4) A linear relationship between DTMC and biodegradable organic carbon has been observed, the gradient of this linear correlation, however, depended on the source of reclaimed water. PMID- 10842828 TI - Selection criteria for wastewater treatment technologies to protect drinking water. AB - The protection of water bodies used as sources for drinking water is intimately linked to the adoption of adequate technologies for the treatment of the wastewater generated in the catchment area. The paper presents a general overview of the main technologies used for the treatment of domestic sewage, with a special emphasis on developing countries, and focussing on the main parameters of interest, such as BOD, coliforms and nutrients. A series of tables, figures and charts that can be used for the preliminary selection of treatment technologies is presented. The systems analysed are: stabilisation ponds, activated sludge, trickling filters, anaerobic systems and land disposal. Within each system, the main process variants are covered. Two summary tables are presented, one for quantitative analysis, including easily usable information based on per capita values (US$/cap, Watts/cap, m2 area/cap, m3 sludge/cap), and another for a qualitative comparison among the technologies, based on a one-to-five-star scoring system. The recent trend in tropical countries in the use of UASB (Upflow Anaerobic Sludge Blanket) reactors is also discussed. PMID- 10842829 TI - Upgrading wastewater treatment plants. AB - Instead of construction of new wastewater treatment plants upgrading of existing plants may increase capacity or achieve higher efficiency in meeting effluent standards. This paper describes several possibilities of upgrading existing technical wastewater treatment plants, e.g. activated sludge plants, e.g. by precipitation/flocculation processes, increase of biomass concentration, influent balancing, increase of oxygenation capacity with pure oxygen, increase of the capacity of final clarifiers, and pretreatment of industrial effluents. PMID- 10842830 TI - Constructed wetlands as a tool for sewage and stormwater pollution control. AB - Although traditionally surface water pollution control measures in many countries of the world have been restricted to sewage effluent discharges, now it has become necessary to apply control measures to stormwater pollution as well. Constructed wetlands are considered as natural systems that provide a low cost maintenance and low energy natural treatment option for sewage and stormwater. They are especially suited for polishing effluent from existing or new sewage treatment plants, or providing treatment for septic tank effluent, by removing organic matter, suspended solids, nutrients, pathogens and heavy metals. This paper describes some important issues relating to the design of constructed wetlands and provides a summary of a pilot study with constructed wetlands for sewage treatment. PMID- 10842831 TI - The use of reed plants for wastewater treatment: the Iraq experience. AB - The use of plants, especially of species that grow naturally and under harsh environmental conditions, offers a simple and economic method of wastewater treatment. This method has been gaining wide popularity throughout the world over the last decades. Iraq is considered to be a country with environmental conditions favourable for the natural growth of many aquatic plants such as reed (Qassab). The root-zone method of treating wastewater is suitable for Iraq, e.g. because of its economy and the fact that such treatment does not need sophisticated mechanical or electrical equipment, or chemical materials. The most important aspect is that this method makes use of a weed, namely the reed. A mobile test tank for root-zone treatment stationed in the Rustamya sewage treatment plant and run for nearly 34 weeks. A total of 768 samples of the influent and effluent sewage were analysed. Average removal efficiencies for biological oxygen demand, suspended solids, chloride, phosphate and ammonia ranged from 92 to 82%, and chloride was removed with 13% efficiency. These results indicate that the root-zone method is adequate for the treatment of domestic sewage and can compete with conventional treatment methods whose average effluent concentrations for BOD5 and suspended solids are similar. A survey of water quality was conducted on a drainage ditch north-west of Baghdad to assess its natural purification process. The ditch that was chosen was infested with reed plants and other aquatic weeds. The role of the macrophytes was found to be essential in the process of self-purification of the water body. Results of the survey indicate that such ditches and streams could be used as disposal points for domestic sewage and industrial wastes. However, further assessment of their performance particularly for removal of suspended solids, pathogens, phosphate and nitrogen is necessary. The results indicate that reed beds may be suitable particularly for treatment of wastewater from small and remote towns and villages in Iraq. PMID- 10842832 TI - Membrane-supported biological wastewater treatment. AB - Research for a feasible method of complete pathogen removal from municipal wastewater has now arrived at a new stage of biological sewage treatment. Microfiltration membranes are integrated into the biological stage where they replace conventional sedimentation for the separation of treated water from sludge. These membranes form a physical barrier that even particles as small as bacteria cannot penetrate. Thus, the biological stage can be operated at higher biomass concentrations, which leads to better treatment performances and drastically reduces the excess sludge production. Results of test runs with pilot plants and municipal sewage suggest that the degradation and elimination potential of optimised membrane bioreactors will result in effluent concentrations as low as natural background concentrations of surface waters. Microfiltration membranes with a pore-size of 0.2 micron or less do not only retain bacteria but also viruses almost completely, and the clarified wastewater is practically free of pathogens. In consequence of these effluent qualities a much higher rate of wastewater can potentially be reclaimed. This paper includes results gained at a pilot plant operated at the Institute for Water, Soil and Air Hygiene of the Federal Environmental Agency in Berlin. PMID- 10842833 TI - Improved purification of water and waste-water with low-cost dead-end ultrafiltration. AB - One aspect of the increasing contamination of water sources is sewage effluent. In some countries it is generally disinfected prior to discharge to protect downstream communities, which use the water for drinking and recreation. However, serious questions have been raised regarding the efficacy of conventional treatments to remove or destroy viruses, and regarding the generation of harmful byproducts by sewage chlorination. A safe and reliable solution is the use of ultrafiltration for the purification of wastewater, as the ultrafiltration membranes form an absolute barrier for bacteria and viruses including colloids and macromolecules. However, this application also demands the use of open channel module systems that can be cleaned with high efficiency with regard to scaling, fouling and especially biofouling. The flat membrane module systems using selected membranes and a special plant design can meet these and further requirements expected in this application, including easy handling, low energy consumption and optimised operation costs. Technical details of the module system, case studies and cost aspects are presented. PMID- 10842834 TI - Sewage disinfection towards protection of drinking water resources. AB - Wastewater applied in agriculture for irrigation could replace the use of natural drinking-water resources. With respect to high concentrations of human pathogens wastewater has to be disinfected prior to use. This paper introduces disinfection methods with emphasis on UV irradiation. PMID- 10842835 TI - Sewage sludge treatment with lime. AB - The article describes the application of lime as a method for treatment and hygienisation of sewage sludges with lime products such as quicklime, slake lime and dolomitic lime. As a result of the increase in temperature and pH-value during sludge and lime mixing most pathogenic vectors of disease (i.e. bacteria, worms, viruses and parasites) are reduced in concentration and viability to manufacture a safe product for further application on agricultural land. PMID- 10842836 TI - Experimental model of faecal coliform removal in anoxic wastewater stabilisation ponds. AB - Wastewater stabilisation ponds are a good alternative for treating different types of wastewaters due to their simplicity, effectiveness and low cost. It has been found that the volumetric organic loading (VOL) on ponds affects pathogen removal inversely whereas temperature has a positive effect. This was confirmed in the present studies on "anoxic" ponds, which were loaded in the range between conventional organic loading levels for anaerobic and facultative stabilisation ponds. In addition, other environmental factors (light, pH, dissolved oxygen (DO) concentration, and oxidation-reduction potential (ORP) were also found to influence pathogen removal in the model ponds. The effects on faecal coliform removal of volumetric organic loading and temperature are represented in the form of a multiple regression model. The equation developed was found to apply at a significance level of 95% with p < 0.001. PMID- 10842837 TI - A small pit emptying machine: an appropriate solution for a Nairobi slum. AB - This article describes the results of the trial period of a pedestrian-controlled pit emptying machine. This 'vacutug' has been tested for UNCHS (Habitat) in a low income settlement in Nairobi, Kenya, through a local NGO. The trial confirmed the viability of the principle of the vacutug as the machine has been in operation for two and a half years. Repairs have been made locally out of income from the service, spare parts can be obtained and demand for the service is high. Although the cost of the service has already been raised, demand has not been reduced. The technical improvements which need to be made can be incorporated in the design, and the next batch of machines is being constructed for further trials. Management of the service is critical for financial sustainability and even during the trial of the technology, the management aspects have been the most difficult of the service. PMID- 10842838 TI - On-site sanitation: when the pits are full--planning for resource protection in faecal sludge management. AB - In urban centres of industrializing countries, the majority of houses are served by on-site sanitation systems such as septic tanks and unsewered toilets. The faecal sludges (FS) collected from these systems are usually discharged untreated into the urban and peri-urban environment, posing great risks to water resources and to public health. Contrary to wastewater management, the development of strategies to cope with faecal sludges, adapted to the conditions prevailing in developing countries, have long been neglected. The authors describe the current situation in FS management and discuss important issues regarding FS management. Further, treatment options are presented which may prove sustainable in industrializing countries. The objective is to make planners and decision-makers aware of the challenges posed by faecal sludges and of the need to include FS management in strategic planning for urban sanitation improvements. PMID- 10842839 TI - Volta watertight VIP latrine. AB - The Danish International Development Agency (DANIDA) is assisting the government of Ghana to provide potable water and improved sanitation facilities to about 50% of the Volta Region's population over a 10-year Community Water and Sanitation Programme (CWSP). Sanitation options include designs for household Ventilated Improved Pit (VIP) latrines. However, areas along the coastal belt of the region have a high water table, between 1-1.5 m (3.3-4.95 ft). In addition, about 80% of the inhabitants in these areas depend on shallow hand-dug wells for domestic purposes and irrigation of vegetable gardens. With the construction of these sanitation options under promotion, pollution of the groundwater is inevitable. Circumventing the problem by the programme led to the modification of the present designs into the "Volta Watertight VIP Latrine (VVIP)". The design considerations and the components that have been incorporated to enable it function effectively are presented in this paper for discussion. Five (5) demonstration units were built, and used for a year. Used pits were covered and pit contents examined after a period of six months. Observations made at the end of this fallow led to the conclusion that the VVIP could be adapted for high water table areas to prevent groundwater pollution, baring any constructional defects and inefficiencies. PMID- 10842840 TI - Evaluation of the oestrogen-receptor activating potency of environmental samples by means of yeast oestrogen assays. AB - A bioassay based on genetically modified yeast strains was applied to screen environmental samples for their oestrogen-like activity. The yeast system was selected from the vast number of "oestrogen bioassays" available because it permits rapid and easy handling at low costs. The test protocol developed allowed the examination of dimethyl sulfoxide-dissolved samples within one day, with an EC50 between 1.5 nM and 3.5 nM 17 beta-oestradiol and a detection limit between 0.3 nM and 0.5 nM 17 beta-oestradiol for yeast strain I. Advantages and disadvantages as well as future prospects of this kind of oestrogen bioassay will be considered by discussing selected results. PMID- 10842841 TI - Rural sanitation problems in Uganda--institutional and management aspects. AB - Rural Uganda faces a lot of problems caused by poor sanitation facilities such as pollution of water sources, a high rate of waterborne diseases, high expenditures on curative health care, and the threat of reduced educational performance of children through illness, early school drop out, especially of girls. Limited budgets and expenditures for the health sector, lack of staff, lack of accountability and transparency are important factors affecting sanitation status on the national level. Other restrictions can be found at the community level, e.g. taboos, cultural and customary beliefs, ignorance, poverty, or in soil conditions. To address the poor level of sanitation, the Government of Uganda has set up both a whole string of laws and guidelines and an institutional and management framework. One main emphasis was placed on the Participatory Hygiene and Sanitation Transformation Programme (PHAST) introduced in 1994 and since then adopted by several non-governmental organisations (NGO's). PMID- 10842842 TI - Review of the wastewater situation in Morocco. AB - Recent estimations of the wastewater production of Morocco amounted to 370 million m3 per year, and this is expected to increase to 900 million m3 by the year 2020. In most cases wastewater is discharged directly into the environment, either to the sea via short outfalls or onto farmland for irrigation or infiltration. Major improvements in the quality of wastewater are needed urgently because of the strong migration of the rural population towards the towns and the very rapid demographic expansion. Studies for Sanitation Master Plans for the main towns are currently in progress and are a first step towards meeting these requirements. Development of a national master plan for liquid sewage is a way of extending this procedure over the whole territory. PMID- 10842843 TI - An assessment of septage and faecal sludge discharges into surface water sources in Ghana. AB - Surface water abstraction from rivers is the main source of potable water for many of the district capitals, municipal and metropolitan towns of Ghana. The points of abstraction on these rivers are usually located several kilometres downstream of their sources. These rivers serve as sinks for both non-point and point sources of pollution. The reliance of the urban population along these rivers on ill-maintained on-site public toilets and overflowing septic tanks renders urban run-off a vehicle for bacterial contamination of human origin. Downstream villages and small towns along the rivers use the water for drinking and other domestic purposes and therefore incidence of waterborne diseases of bacterial origin is therefore prevalent. This paper discusses the impact of discharges on Densu and Oda, two examples of surface waters employed for urban water supply, and makes suggestions for integrated water quality management as a tool for protecting surface water sources. PMID- 10842844 TI - Ground water in Nigerian urban centres: problems and options. AB - Results of a groundwater study in Nigeria in some of the major cities in the southern part, Lagos, Ibadan; in the eastern part, Warri, Benin, and Aba, and in northern part, Kano and Jos indicates that the quality of waters fell far below the WHO recommended levels for some of the quality parameters. The levels of nitrate, lead and coliform index were particularly far above the WHO limits, and some of the waters have higher levels of iron with low pH values. The quality is poor in high density or low-income areas. In addition Lagos waters also showed intrusion of salinity in the localities closer to the coast. Poor ground water quality was attributed to intrusion of sullage or gray water, indiscriminate defecation, and dumping of household refuse, industrial and hazardous wastes around the premises. Lead from locally produced gasoline and heavy road traffic in the urban centres, seepage of various leachates, inadequate governmental policies and poor implementation strategies, and indifferent attitudes of communities to the environment are some of the other identified reasons. PMID- 10842845 TI - Health risks of rural water supply due to lack of proper sanitation in southeast Asian countries. AB - A comparative study on the effects of lack of sanitation and inappropriate waste handling on water supply was carried out in several Southeast Asian countries that have problems such as inadequate water resources, water supply and sanitation. Though the degree and the cause varied in each country, all the countries investigated had problems arising from contamination of water resources. It was found that there are several sources of water contamination, such as human and animal excreta, solid wastes from nearby houses, graveyards, contaminated river flows, and large-scale landfills for big cities. It was revealed that even when water is clean at the point of production, it could be easily contaminated through broken pipelines or improperly maintained containers. It is still rare to put chlorine into piped water supply and both the inhabitants' and water engineers' understanding of the importance of disinfection should be reemphasised. Although it is urgent to provide piped water supply to those who have only contaminated water sources, such as surface water and dug wells, it is also important to protect these limited water sources from the above mentioned contamination. PMID- 10842846 TI - Status of domestic wastewater management in relation to drinking-water supply in two states of India. AB - In India, supply of drinking water, treatment and disposal of domestic wastewater including faecal matter are managed by local bodies. The existing status of water supply, characteristics of domestic wastewater, modes of collection, treatment and disposal system for sewage and faecal matter in 82 municipalities and 4 municipal corporations were assessed in the States of Bihar and West Bengal in India. Domestic wastewater in the municipal areas is collected and discharged through open kachha (earthen), pucca (cement-concrete) and natural drains and discharged into water courses or disposed on land. Scavenger carriage system for night soil disposal is in-vogue at several places in the surveyed States. Open defecation by the inhabitants in some of the municipalities also occurs. The existing methods of collection, treatment and disposal of sewage impairs the water quality of different water sources. Techno-economically viable remedial measures for providing basic amenities, namely safe drinking-water supply and proper sanitation to the communities of these two States of India are suggested and discussed. PMID- 10842847 TI - Pollution of water sources and removal of pollutants by advanced drinking-water treatment in China. AB - The pollution of water resources and drinking water sources in China is described in this paper with basic data. About 90% of surface waters and over 60% of drinking water sources in urban areas have been polluted to different extents. The main pollutants present in drinking water sources are organic substances, ammonia nitrogen, phenols, pesticides and pathogenic micro-organisms, some of which cannot be removed effectively by the traditional water treatment processes like coagulation, sedimentation, filtration and chlorination, and the product water usually does not meet Chinese national drinking water standards, when polluted source water is treated. In some drinking-water plants in China, advanced treatment processes including activated carbon filtration and adsorption, ozonation, biological activated carbon and membrane separation have been employed for further treatment of the filtrate from a traditional treatment system producing unqualified drinking water, to make final product water meet the WHO guidelines and some developed countries' standards, as well as the Chinese national standards for drinking water. Some case studies of advanced water treatment plants are described in this paper as well. PMID- 10842848 TI - Sludge treatment and disposal in Slovakia. AB - The ongoing economic changes that accompany the transformation period of Central and Eastern European (CEE) countries have significantly affected the production of wastes generated in urban areas and/or particularly in municipal wastewater treatment plants (WWTPs). The production of sewage sludges is expected to increase in Slovakia because of upgrading the existing plants, requiring more extensive sewage treatment, and because of construction of new treatment plants. Analysis of available data combined with the latest results of comprehensive monitoring of sludge generated at municipal WWTPs shows the serious problems concerning its quality and thus in general with sludge management and its reuse. Based on results obtained from monitoring, an amendment of the present legislation for sludge disposal in agriculture is under preparation. Priorities of state environmental policy should include improvement of the present situation is sludge treatment and disposal in Slovakia. PMID- 10842849 TI - New policies and measures for saving a great manmade reservoir providing drinking water for 20 million people in the Republic of Korea. AB - Water quality of the Paldang reservoir, the largest drinking water supply source in the Republic Korea provides raw water for about 20 million people living in Seoul Metropolitan area. Water quality has been deteriorating mainly due to improperly treated livestock waste and domestic wastewater discharged from motels, restaurants, and private homes. A recent survey conducted by the Ministry of Environment (MOE) showed that the water quality of this reservoir has been identified as Class III must contain less than 6 ppm of BOD, which will require advanced purification treatment before it can be used as drinking water. The MOE also announced that this water source would no longer be potable unless wastewater in the catchment is treated efficiently. To protect drinking water resources, the MOE has set up comprehensive management. These programmes include new regulations, measures, land use planning and economic incentives. PMID- 10842850 TI - The Cyprus experience with protection of ground- and surface waters from domestic sewage and excreta. AB - Water resources in Cyprus are scarce and expensive to exploit; rainfall is highly variable and droughts occur frequently. The Cyprus authorities are concerned with the conservation and protection of water supply sources. For this purpose the Water Pollution Control Law has been issued. According to the Street and Buildings Law, all dwellings must be equipped with a septic tank, followed by an absorption pit. When the pits overflow due to saturation of the soil, the septage is pumped out and transported to a sewage treatment plant for treatment and reuse. Based on land-use zoning, housing developments are not allowed in the vicinity of water-supply sources, rivers and reservoirs. In order to avoid contamination of the water sources from sewage and excreta, protection zones are designated in which the disposal of sewage is not allowed. PMID- 10842851 TI - Treatment news: 6th Conference on Retroviruses and Opportunistic Infections. PMID- 10842852 TI - Partner notification as a prevention strategy: a social system perspective. AB - Provider referral and contact referral are two established means to provide notification of contagious, but treatable diseases, as has been done for tuberculosis and sexually transmitted diseases (e.g., syphilis). Because AIDS is at this time an incurable disease, programs of notification have proved highly controversial. The author examines recent adjustments to the New York State public health law regarding HIV/AIDS notification mechanisms. A literature review supports further discussion of ethical and partner notification issues as they bear on the perception and objections of various constituencies, stigmatization, and principles of confidentiality. Both U.S. and Swedish strategies to partner notification are outlined as well as why these schemes may not be interchangeable based on culture differences; points for general improvement are drawn from this comparison. A social system perspective based on the social functions of adaptation, goal attainment, integration, and latency (pattern maintenance) is introduced as a means to address the barriers inherent in HIV/AIDS notification programs and to enhance counseling programs--the objective being that knowledge and understanding of the patient's culture and social context can give providers additional tools with which to stop the spread of HIV/AIDS and bring people to treatment earlier. PMID- 10842853 TI - Coping styles of caregivers of children with HIV/AIDS: implications for health professionals. AB - In a study to examine the coping styles and quality of life of non-parental caregivers of children with HIV/AIDS, 25 non-parental caregivers used confrontive coping most often and passive and emotive coping much less often. The two most frequently used coping styles were prayer and efforting to maintain some control over the situation. Quality of life correlations showed that support from family/friends was significantly related to all of the quality of life subscales. Time spent caring for the HIV-positive child was significantly inversely related to all aspects of quality of life. The conclusions drawn from the study were that interventions are needed to help support caregivers of children with HIV/AIDS. Examples of primary, secondary, and tertiary prevention measures for health professionals are discussed. PMID- 10842854 TI - Gastrointestinal hemorrhage in patients with AIDS. AB - Gastrointestinal (GI) bleeding is a relatively infrequent complication seen in patients with AIDS. As with non-HIV-infected individuals, upper GI bleeding is much more common than lower GI bleeding. In patients with AIDS, upper GI bleeding can result from etiologies related to underlying HIV infection [cytomegalovirus (CMV), Kaposi's sarcoma, idiopathic esophageal ulcers, etc] or be unrelated to HIV infection (peptic ulcer, portal hypertension, Mallory-Weiss tear, etc.). Lower GI bleeding is caused predominantly by etiologies related to underlying HIV disease; CMV colitis is the most common cause. In contrast to non-HIV-infected individuals, hemorrhoids and anal fissures can result in significant bleeding in AIDS patients because of associated thrombocytopenia. Management of GI bleeding in AIDS patients is similar to patients without HIV infection, and includes resuscitation, identification of the bleeding source, achieving hemostasis, and preventing recurrent bleeding. Several etiologies that cause GI bleeding in patients with AIDS can be diagnosed through endoscopy, either by their characteristic endoscopic appearance or mucosal biopsies. PMID- 10842855 TI - Changing spectrum of HIV infection and its associated conditions in Spain: the end of the beginning? AB - To evaluate the natural history and the impact that different strategies have had on HIV infection and its associated conditions, a cohort of 970 patients (432 had AIDS) who had been seen over a period of 13 years were studied. The incidence of new HIV-infected patients had increased steadily since 1985, peaking in 1993 (52.9 cases/100,000 population), to significantly decrease in the subsequent years. The most common AIDS-defining illnesses (ADIs) were tuberculosis (52.3% of the patients), Pneumocystis carinii pneumonia (30%), and candidal esophagitis (28.6%). The frequency of new cases of tuberculosis per patient with ADI clearly decreased over this period (p < 0.0001), whereas that of P. carinii pneumonia decreased slowly until 1994, to fall thereafter to about half the previous levels (p = 0.005). Candidal esophagitis showed a biphasic pattern, the second peak probably due to the emergence of fluconazole resistance. The number of ADIs per patient increased from the beginning of the observation period, peaking in 1995 (1.67 ADI/patient), followed by a statistically significant decrease that, in 1997, reached the 1987 levels (1.22 ADI/patient). We conclude that the clinical spectrum of HIV infection is improving in the last years. Except for tuberculosis, prophylaxis for other ADIs, although effective, does not seem to have had a dramatic impact on occurrence. The most impressive reductions have been observed only after the introduction of antiretroviral combination therapy. PMID- 10842856 TI - Determinants of HIV-related survival among Texas prison inmates. AB - Research indicates that being incarcerated adversely affects disease progression and overall health status. Because HIV infection is a growing problem among prison populations in the United States, understanding how incarceration affects HIV-related survival patterns is critical. The present study examined determinants of HIV-related survival in a cohort of 2380 Texas Department of Criminal Justice (TDCJ) inmates who were treated for HIV/AIDS, dating from January 1, 1992 and June 31, 1997. Assessment of the study factors indicated that there were no substantial violations of the assumptions of the Cox's proportional hazards (PH) model in the present study population. Furthermore, to address the potential problem of censoring-related bias, mortality information was collected on all inmates who were paroled on the basis of disease status. The present study's findings indicate that the following factors were associated with significant decreases in HIV-related survival in the TDCJ prison population: male gender, older age, self-report of no known HIV transmission risk factors, and presence of cytomegalovirus (CMV), Mycobacterium avium complex (MAC), and Pneumocystis carinii pneumonia. Moreover, survival decreased in a monotonic fashion with decrease in baseline CD4 count. While the majority of the present study's findings were consistent with those reported for non-incarcerated populations, it will be important for investigators to assess whether these findings persist among future cohorts of prison inmates. PMID- 10842857 TI - Return rates and partner notification in HIV-positive men seeking anonymous versus confidential antibody testing. AB - This study investigated whether HIV-positive men who seek confidential versus anonymous HIV counseling and testing differ in demographic variables, risk behaviors, return rates for posttest appointments, and agreement to partner notification. chi 2 tests were not statistically significant for return rates for post-test appointments or partner notification between the two groups. HIV positive individuals in the confidential groups were more likely to utilize medical and follow-up services than those in the anonymous group. Anonymous and confidential counseling and testing both appear to achieve the public health objectives of HIV case finding and referral. Hypotheses are offered regarding what may be a progression of testing behaviors (i.e., from anonymous to confidential) with suggestions for future research are suggested. PMID- 10842858 TI - Periodontopathic bacteria in English HIV-seropositive persons. AB - Selected periodontopathic bacteria were sought in 20 HIV-infected English patients and eight noninfected control subjects with similar periodontal status, using highly specific DNA probes. Actinobacillus actinomycetemcomitans (A. a), Porphyromonas gingivalis, Campylobacter rectus, Prevotella intermedia, and Fusobacterium nucleatum were more frequently detected and were found at higher levels in HIV-infected individuals than in controls. Significantly increased levels of Treponema denticola but, in contrast, lower levels of Eikenella corrodens were found in nonbleeding sites of HIV-infected patients compared with controls. The results of the present investigation on English patients suggest a periodontopathogenic role for A. a, P. gingivalis and C. rectus, and possibly P. intermedia and are in general accord with most previously reported data from the United States. Longitudinal studies are now required to determine more precisely the association between periodontopathic microflora, immune competence and periodontal health and disease in HIV-infected persons. PMID- 10842859 TI - Drug holidays may be beneficial. PMID- 10842860 TI - Recent antiviral approvals. PMID- 10842861 TI - HE2000 shows efficacy. PMID- 10842862 TI - Sexual dysfunction with PIs. PMID- 10842863 TI - Vaccine trial in Thailand. PMID- 10842864 TI - Teen STD screening in St. Louis. PMID- 10842865 TI - Test detects HPV. PMID- 10842866 TI - Plant may be cure for HBV. PMID- 10842867 TI - No vaccine for 10 years. PMID- 10842868 TI - TB continues to decline. PMID- 10842869 TI - Youth believe pill stops HIV. PMID- 10842870 TI - Hepatitis rises in Texas. PMID- 10842871 TI - European approaches to adolescent sexual behavior. PMID- 10842872 TI - ASHA kicks off herpes education campaign. PMID- 10842873 TI - Treatment of bone and joint infections: recommendations of a Belgian panel. AB - A multidisciplinary panel of Belgian specialists describes the overall therapeutic approach for bone and joint infections. Classification, general methods of investigation, therapeutic options, special circumstances, the role of aminoglycosides and of glycopeptides are described. The possibility of home treatment is discussed, as well as some pharmaco-economic insights. PMID- 10842874 TI - Congenital insensitivity to pain with anhidrosis. Report of a case and review of the literature. AB - In a previous paper published in this journal, we reported two cases of "Congenital Sensory Neuropathy with Anhidrosis" with reference to the orthopedic complications (Theodorou et al., 1985). We now present a new typical case, under the currently used term: "Congenital Insensitivity to Pain with Anhidrosis" (CIPA) and a brief review of the literature on the incidence, etiology and problems arising in various systems. CIPA is an autosomal recessive form of sensory neuropathy manifesting with typical clinical features. Universal insensitivity to pain, anhidrosis or hypohidrosis, bouts of hyperpyrexia from very young age, self inflicted injuries, defective or absent lacrimation and mental retardation are specific diagnostic findings. Orthopedic, maxillofacial, dermatological and ophthalmologic complications are common. Counseling of the family and school personnel for the prevention of injuries is necessary. Early diagnosis is very important for the prevention and treatment of various complications. The etiology and pathogenesis of the condition is still unclear. The recent detection of a new gene, which encodes a receptor tyrosine kinase for nerve growth factor and lately of a specific point mutation associated with the gene inactivation11, may open new ways for the study and management of this disabling condition. PMID- 10842875 TI - [Solitary osteochondroma of the limbs. Clinical review of 76 cases and pathogenic hypothesis]. AB - The authors report a retrospective study of 76 solitary osteochondromas of the appendicular skeleton treated between 1981 and 1995. The ages of the patients ranged from 13 to 55 years with a mean of 21 years. The male/female-ratio was 1.37. Tumefaction with or without associated pain was the reason for consultation in 68 cases (89%). In 62 cases the osteochondroma was localized in the distal part of the femur or the proximal part of the tibia. All lesions were surgically resected; the resection was complete in all cases. Six patients were lost to follow-up; the other 70 were seen on a regular basis over a time period ranging from 1 to 12 years. The result from surgical treatment was assessed based on pain, joint motion, cosmetic consequences, nerve compression and recurrence of osteochondroma. The results were good in 68 cases and fair in two cases. Based on a review of previous experimental studies, the authors suggest a hypothesis to explain the rotation of a fragment of the growth plate which is needed for the development of osteochondroma. This rotation occurs as a result of the mechanical action from the periosteum under tension. PMID- 10842876 TI - [Fractures of the trapezium]. AB - The authors report a retrospective study of eight cases of trapezium fractures. There were seven male and one female patient aged 21 to 48 years. The presumed mechanism of injury was established in only four cases. Clinical trapeziometacarpal abnormality was always present. The diagnosis was made on standard radiographs in seven cases, computed tomography was used in one case. The fracture was simple, vertical and lateral in five cases, complex in one case and consisted in a lateral avulsion in two cases. An associated Bennett's fracture was present in five cases. The treatment was conservative in two cases. Internal fixation was performed in four cases (3 using a lag screw, 1 using a Kirschner wire) and transosseous reinsertion in two cases. With an average follow up of 3 years (1 year-6 years), open reduction and internal fixation appears to be simple and to give good or excellent results. PMID- 10842877 TI - The role of osteoporosis in distal radius fractures. AB - Our study was designed to establish whether the degree of osteoporosis of the distal forearm affects the outcome of distal radius fractures in elderly women. We assessed the Gartland and Werley score, wrist mobility, grip strength, ulnar variance, radial inclination and palmar tilt of both wrists in 27 postmenopausal women who had sustained a unilateral distal radius fracture following a simple fall. Bone mineral density of the contralateral uninjured wrist and mid-tibial ultrasound velocity were measured. The Gartland and Werley score, wrist mobility, loss of grip strength and the radiological results of the fractured wrist did not correlate with bone mineral density of the uninjured distal radius or with mid tibial ultrasound velocity. These results may indicate that the influence of osteoporosis on the radiological and clinical outcome in distal radius fractures is not very important. PMID- 10842878 TI - Glomus tumors. AB - The authors report a series of 12 consecutive patients with 13 glomus tumors operated from July 1991 until February 1999. Symptoms were present for an average of 1.9 years before surgery. Women were more frequently affected. The mean age was 44 years. In 12 of the 13, the tumor was located in the distal phalanx and one patient had a glomus tumor on the dorsum of the hand. One glomus tumor was found in the right hallux of a two-year-old child. Both hands and all fingers were equally involved. One bilateral glomus tumor was associated with neurofibromatosis. All tumors were resected and histology confirmed the diagnosis. The result was good with immediate pain relief. No recurrence has been noted to date. PMID- 10842879 TI - Heterotopic ossification in total hip arthroplasty: the significance for clinical outcome. AB - This study evaluates 706 patients with 835 primary total hip replacements documented in a prospective fashion in a multicenter study with respect to correlation between heterotopic ossification (HO) and clinical outcome. Only patients without prophylaxis against HO entered the study. The mean clinical and radiological follow-up was 3.1 years (+/- 0.7). Heterotopic ossification was noted in 47% of all total hips replaced. It was graded as mild (Brooker I) in 29.1%, moderate (Brooker II) in 12.7%, and severe (Brooker III and IV) in 5.2%. All clinical parameters investigated were significantly affected with the increasing amount of heterotopic ossification. The strongest correlation was found in flexion range and spreading distance. Both factors were significantly decreased with higher degrees of ossification. The other clinical parameters investigated, walking capacity, limp, and use of analgesics, were altered to a lesser extent and only with higher degrees of heterotopic bone formation. Finally, patient satisfaction was significantly influenced by the degree of heterotopic ossification and dropped from almost 90% good or excellent patient satisfaction in the non-ossification group to less than 30% in the group with severe ossification. PMID- 10842880 TI - Subtrochanteric valgus-extension osteotomy for neglected congenital dislocation of the hip in young adults. AB - Thirty-five patients with unilateral or bilateral neglected congenital dislocation of the hip (CDH) were treated with subtrochanteric valgus-extension osteotomy between 1975 and 1992. There were 29 females and 6 males in the group. A total of 50 osteotomies was performed. The mean age of the patients was 22 years; the mean follow-up was 7 years. Before operation, the main complaints were pain and gait abnormalities. Leg-length discrepancy was another problem, especially for unilateral cases. The main indication for the operation was pain. This pelvic support osteotomy was performed to correct the instability of the hip and as a result of this to relieve pain. Patients were retrospectively evaluated based upon Harris Hip Score and self-evaluation. The mean Harris Hip Score was 49 before operation and improved to 72 after the operation. Alleviation of the pain was the most significant functional outcome of the treatment. It was also noted that limping could be improved if an adequate rehabilitation program was followed. We conclude that in neglected CDH cases with pain, limping and lumbar hyperlordosis in the young adult, a subtrochanteric valgus-extension osteotomy can give satisfactory results. PMID- 10842881 TI - Recurrent traumatic dislocation of the hip joint at the age of 13 and 17 years. A case report. AB - We report a case of a recurrent traumatic dislocation of the hip joint in a young man at the age of 13 and 17. The patient had retroversion of the femoral neck, instead of a physiological anteversion. We found no consensus about the management of a recurrent traumatic dislocation of the hip joint in the literature. The various therapeutic options are discussed. We advised posterior capsulorraphy with femoral rotation osteotomy. The patient refused surgery. PMID- 10842882 TI - [Lumbar Chance fracture in an child associated with an intra-abdominal lesion]. AB - We report the case of a teenager who was involved in a road traffic accident. She presented a flexion-distraction type of vertebral injury, (Chance fracture). This fracture was associated with an intra-abdominal injury. The child was a passenger in a rear seat using a shoulder seatbelt restraint. PMID- 10842883 TI - Intraosseous ganglion of the triquetrum. A transpisiformal approach. AB - A cystic subchondral bone defect without joint pathology is called an intraosseous ganglion. Most occur in the lower limb. In the wrist the scaphoid and lunate are most often involved. We report a case of an intraosseous ganglion within the triquetrum, treated by curettage and grafting with the pisiform that had been removed. PMID- 10842884 TI - Bifocal pubic stress fracture after ipsilateral total knee arthroplasty in rheumatoid arthritis. A case report. AB - We present the case of a 78-year-old rheumatoid patient with a bifocal stress fracture of the pubic rami after ipsilateral total knee arthroplasty. Many risk factors that may cause insufficiency fractures are combined in rheumatoid arthritis. These patients are used to chronic pain, and therefore the diagnosis is often made late. When a patient suffering from rheumatoid arthritis presents with recent pain in the pelvic or hip region after hip or knee arthroplasty, a stress fracture should always be suspected. A standard pelvic radiograph and skeletal scintigraphy establish the diagnosis. CT-scan may be useful in the differential diagnosis with less benign pathology. Rest and analgesic drugs are sufficient to let the fracture heal. Nonunion has never been reported for these fractures. PMID- 10842885 TI - The assumption of the supportive function by the fibula as a consequence of acquired untreated tibial pseudarthrosis. AB - The authors present a case of acquired, untreated pseudarthrosis of the tibia which resulted in adaptation to the weight-bearing function by the fibula. The case described shows the adaptation abilities of the bone tissue which, when subjected to unusual loading, undergoes the necessary reconstruction. PMID- 10842886 TI - [Tibialization of the fibula for a large bone loss. A case report]. AB - A 10-centimeter tibial bone loss was treated by inter-tibiofibular bone grafting on each part of the pseudarthrosis, resulting in tibialization of the fibula. The patient was reviewed after 10 years; the clinical result was satisfactory and stable. The tibial forces were deviated on the fibula, which expressed radiologically by important thickening of its cortices. This case illustrates the interest of preserving the fibula for tibial bone loss reconstruction. PMID- 10842887 TI - [Angiogenesis and anti-angiogenesis in human neoplasms. Recent developments and the therapeutic prospects]. AB - Angiogenesis entails new vessel formation from preexisting vessels. It follows vasculogenesis during embryo development. In post-natal life, it occurs both in physiological conditions (wound repair and cyclically in the female genital system) and pathological conditions such as tumors. Several sequential steps are involved, including basement membrane degradation by proteolytic enzymes secreted by the endothelial cells, chemotaxis toward the stimulus and proliferation of these cells, canalization, branching and formation of vascular loops, stabilization and functional maturation of neovessels following perivascular apposition of pericytes and smooth muscle cells, and neosynthesis of basement membrane constituents. Tumor angiogenesis is regulated by several factors, mainly growth factors for the endothelial cells secreted by both the tumor and host inflammatory cells, and mobilized from extracellular matrix stores by proteases secreted by tumor cells. Regulatory factors also include the extracellular matrix components and endothelial cell integrins, hypoxia, oncogenes and tumor suppressor genes. Angiogenesis is mandatory to the process of tumor progression (growth, invasion and metastasis), since it conveys oxygen and metabolites, whereas endothelial cells secrete growth factors for tumor cells and a variety of proteinases which facilitate invasion and increase opportunities for tumor cells to enter the circulation. We present our results concerning the relationship between angiogenesis and progression in patients with melanoma, multiple myeloma, B-cell non-Hodgkin's lymphomas and mycosis fungoides. Lastly, it is becoming increasingly evident that agents interfering with blood vessel formation also interfere with tumor progression. These include antagonists of angiogenic growth factors, angiogenic receptors, endothelial cell integrins, and proteolytic enzymes, as well as non-specific toxic agents for vessels and low-dose chemotherapeutic agents. Their recent applications in preclinical models and in neoplastic patients are reviewed. PMID- 10842888 TI - [HIV-related thrombocytopenias]. AB - Thrombocytopenia is a relatively frequent hematological complication in patients infected with the human immunodeficiency virus type 1. Occurring in all risk groups (homosexuals, intravenous drug addicts, hemophiliacs, heterosexuals) this complication has an overall prevalence varying between 5 and 15%. Only a relatively small proportion of patients have severe thrombocytopenia (6 to 24%), and life-endangering bleeding symptoms are not frequent. Accordingly, aggressive treatment is seldom indicated. Corticosteroids should be avoided because they may further compromise the deteriorated immune system of these patients. Splenectomy is efficacious as in patients with non-human immunodeficiency virus related immune thrombocytopenic purpura and infectious complications are not a prominent problem following the operation. Antiretroviral treatment with zidovudine and/or highly active protease inhibitors increases platelet count, but the control of thrombocytopenia is often transient when these drugs are stopped. Other treatments such as alpha-interferon and thrombopoietin require further experiences. On the whole, thrombocytopenia is generally a relatively mild complication of the human immunodeficiency virus infection and aggressive treatment is often unwarranted. PMID- 10842889 TI - [The difficulties of translating into daily clinical practice the favorable results of clinical trials in chronic heart failure]. AB - Congestive heart failure is a growing public health problem and continues to be characterized by a poor prognosis. Its prevalence may be estimated at 0.4-2% of the general population with an incidence varying from 1-10% in elderly patients. This may be in part explained by the progressive aging of the population and the more effective therapeutic strategies for coronary heart disease and hypertension: since they increase the life-expectancy of patients, they favor the onset of this clinical syndrome at a more advanced age. The number of hospital admissions has also increased; mortality is 50% within 4 years of diagnosis, although patients with very severe cardiac decompensation die within 1 year. Despite the fact that with the use of the most advanced molecules created by pharmacological research (ACE inhibitors, beta-blockers, angiotensin-II-receptor antagonists) a significant reduction in morbidity and mortality as well as clear improvement of symptoms and quality of life have been obtained in diverse clinical trials, there is still no certain evidence that in the general population, mortality due to chronic heart disease has been reduced. In our opinion, several factors explain the gap between the favorable results of clinical trials and the lack of significant reduction in morbidity and mortality in clinical practice: A) the selection criteria utilized in clinical studies exclude, for the most part, patients present in general population; B) the average age of the patients included (60 years) is not representative of the overall population; C) patients with associated co-morbidity are generally excluded; D) the number of women enrolled in the trials is quite low; E) compliance is certainly better in the trials than in clinical practice; F) higher drug dosages are used in the trials than in clinical practice. To intervene in these areas and thus make the positive results obtained in the trials available to the general population is, in our opinion, the principal challenge of the future. For this task, the close collaboration and commitment of the internist, the geriatrist, the cardiologist and the family doctors is necessary so that the results reported in clinical trials may be translated into general medical practice. PMID- 10842890 TI - [The limits of controlled clinical studies: the case of rheumatology]. AB - The double blind, placebo controlled, randomized clinical trial is the best tool for gathering evidence concerning the efficacy and safety of a drug. However, in some clinical situations--rheumatology is one--these trials have an intrinsic difficulty in representing the clinical reality: e.g., indicating how a drug influences course of a disease over the long-term. Beginning with the distinction between the "activity" of a drug as confirmed by randomized clinical trials, and its "efficacy" over time, as desired by the patient, we attempt to question why this difficulty arises. Several differentiating elements are involved. The objective of the randomized clinical trial design, that is, activity on acute parameters, may not coincide with the interests of the patient, i.e., efficacy in controlling disability or chronic pain. The duration of a randomized clinical trial is sufficient to measure drug activity but not to provide an indication of its effects on the course of the disease. In the randomized clinical trial, the size of the sample, generally from 10(2)-10(3) patients can, at the most, lead to the conclusion that a drug is "active" while the measurement of efficacy is based on a much greater number of observations. With regard to recruitment criteria, clinical trial patients are chosen on the basis of defined disease severity and activity, while in clinical practice, it is known that this often waxes and wanes. The presence of co-morbidity often leads to the exclusion of patients, yet it is known that up to 17% of rheumatoid arthritis patients are depressed. While concomitant therapy is not permitted during randomized clinical trials, it is generally almost always implemented in normal clinical practice. Drug dosage in a clinical trial is not variable, while in practice it is. The greatest difference is found in the measurement of efficacy, where the randomized clinical trial emphasizes the number of tender and swollen joints, acute phase reactants, disease activity indexes, to assess improvement, while for a typical patient, efficacy is measured in terms of non-evolution of radiologic alterations, work capacity and deformity, and/or the need for a joint prosthesis. The objective of the clinical trial is to seek improvement, while a patient with rheumatoid arthritis may consider simply the lack of worsening over a 5-10 year period as a success. Contrary to clinical trials, observational studies function well in this situation. In response to the difficulty that randomized trials have in reflecting the clinical reality of rheumatological outcomes, the solution is to utilize within the trial, the aspects of the disease course considered fundamental by the patient (that is, chronic pain, disability, radiographic alterations), parameters that are generally omitted from trials. A radical alternative, already proposed, is to abandon the randomized clinical trial model completely and adopt open approaches--much less stringent than randomized trials although as stringent as possible outside the framework of the trial model--that are able to reflect the problems of the patient and to respond to them. PMID- 10842891 TI - [Glucocorticoid-induced osteoporosis: recent findings]. AB - The clinical features, pathogenesis and management of bone involvement in Cushing's syndrome are briefly reviewed. Personal data on bone mineral density and markers of bone turnover in Cushing's syndrome and adrenal incidentalomas are also reported. As long ago as 1932, Harvey Cushing recognized osteoporosis as a serious consequence of endogenous hypercortisolism. The introduction of cortisone in the therapy of autoimmune, rheumatic, allergic or dermatologic disorders was followed by several reports of detrimental effects on bone of patients who had undergone prolonged glucocorticoid treatment. Due to the rarity of Cushing's syndrome, most of the studies in the literature on glucocorticoid-induced osteoporosis refer to exogenous over-exposure to cortisone and its synthetic derivatives. Only a small number of works concern endogenous hypercortisolism, even if the characteristics of bone damage seem qualitatively the same. Finally, very few data are reported on the hypothetical detrimental effect on bone in the condition of the silent hypercortisolism of adrenal incidentalomas. Glucocorticoid-induced osteoporosis in Cushing's syndrome often results in vertebral fractures, and bone loss is more evident in trabecular than in cortical bone. Notwithstanding some distinctive features in osteoporosis induced by endogenous and exogenous glucocorticoid excess, the common eventual picture is notable bone damage that involves mainly the trabecular bone. Prompt and effective therapy is mandatory to reduce the risk of fractures. The present options include calcium and vitamin D supplementation, estrogen replacement therapy, bisphosphonates, either oral or parenteral. A novel approach to the clinical problem of glucocorticoid-induced osteoporosis might, in the future, be based on studies on selective glucocorticoid receptor modulators, a new class of synthetic glucocorticoids that exhibit significant anti-inflammatory and immunosuppressive activities, with reduced side effects on bone. PMID- 10842892 TI - [Primary and secondary autoimmunity in hepatology]. AB - Primary autoimmune liver diseases can be hepatitic or cholestatic in nature. Autoimmune hepatitis, more often diagnosed in women, is characterized by biochemical and histological activity, with polyclonal hypergammaglobulinemia as a frequent feature. Antinuclear and anti-smooth muscle antibodies are the serological hallmarks of type 1 autoimmune hepatitis, whereas liver-kidney microsomal antibody type 1 and liver cytosol antibody type 1 designate the type 2 form. Response to immunosuppression is usually excellent. The most frequent cholestatic autoimmune disease is primary biliary cirrhosis, characterized by anti-mitochondrial antibody positivity and typical bile duct lesions observed on liver biopsy. Treatment with biliary acids improves the biochemical picture, may alleviate pruritus, and delays the development of end-stage liver disease. Primary sclerosing cholangitis occurs more frequently in men and affects both the intra- and extrahepatic biliary trees, determining the typical "beading" appearance. Associated inflammatory bowel diseases are often observed. To date, no medical therapy is able to modify the course of this disease. Autoimmune cholangitis is an anti-mitochondrial antibody-negative cholestatic disease with most of the features of primary biliary cirrhosis. "Overlap" syndromes where autoimmune hepatitic and cholestatic features coexist in the same patient, have also been reported. Autoimmune phenomena secondary to hepatitis C virus-related liver disease such as the occurrence of antinuclear, anti-smooth muscle antibodies and liver-kidney microsomal antibody type 1 are often observed. PMID- 10842893 TI - [The usefulness and limits of the ambulatory monitoring of arterial pressure]. AB - The interpretation of blood pressure values, measured by the physician in the medical environment, has some limitations. Blood pressure fluctuates considerably due to circadian rhythm and various environmental stimuli and thus casual readings may not be representative of the true blood pressure status of the patient. Non-invasive ambulatory blood pressure monitoring has greatly increased our knowledge regarding the diagnosis, treatment efficacy, natural history and even prognosis of hypertension. This technique allows the evaluation of repeated blood pressure measurements at given times (daytime, nighttime, early-morning) and in different conditions (awake, asleep, at rest, during physical or mental activity etc.). Many studies have shown that the whole-day values obtained with such devices correlate better with the morbidity and mortality associated with hypertension than does casual office blood pressure. The 24-hour blood pressure recording is useful in the diagnosis of hypertension, particularly to discriminate so called white coat hypertension (for which the term isolated clinic hypertension has recently been proposed). Ambulatory blood pressure monitoring provides reliable and detailed information on the efficacy of pharmacological treatment and its ability to cover the whole day. These considerations notwithstanding, the application of this technique is still far from routine in the clinical management of arterial hypertension. Its major limit is that the normalcy of ambulatory blood pressure values still remains to be calculated, and only hypothetical normal reference values, derived from studies of selected normotensive populations, are presently available. PMID- 10842894 TI - [The immunomodulatory effect of blood transfusions and intravenous immunoglobulins: the role of the soluble molecules of the Class-I major histocompatibility complex and of the Fas ligand]. AB - Allogeneic blood transfusions may have immunomodulatory effects including improved allograft acceptance and increased risk for cancer recurrence or post operative bacterial infections. These effects are associated with the presence of leukocytes in transfused blood and are reduced by pre-storage leuko-reduction. However, the precise mechanism of this effect has not yet been elucidated. We report that the concentrations of soluble major histocompatibility complex class I and soluble Fas-ligand molecules are significantly higher in supernatants of blood components containing elevated numbers of residual donor leukocytes, such as red blood cells and random-donor platelets, than in other blood components. Elevated amounts of soluble Fas-ligand molecules are also found in some intravenous immunoglobulin preparations. Soluble molecules detected in blood components and in immunoglobulin preparations are biologically active in vitro. In fact, they inhibit mixed lymphocyte responses and cytotoxic T cell activity in allogeneic and autologous combinations and induce apoptosis in Fas-positive cells. These results should be taken into account in clinical practice to select the blood component or the immunoglobulin preparation in order to induce or prevent an immunosuppressive effect in the recipient. PMID- 10842895 TI - [Retinal vein occlusions: diseases for the internist?]. AB - Retinal vein occlusion is a relatively common disease, usually associated with the presence of diseases related to internal medicine. Central retinal vein occlusion is the most frequently-occurring and clinically relevant type. In addition to the well-known classic risk factors, new hemostasis-related ones have been investigated in patients affected by central retinal vein occlusion. While data concerning a number of parameters remain contradictory, high levels of type 1 plasminogen activator inhibitor (PAI-1) and lipoprotein(a) and hyper homocystinemia appear to play a significant role in the pathogenesis of this disease. Although based on a limited number of studies, this new knowledge could eventually provide important indications regarding prognosis and therapeutic strategies. PMID- 10842896 TI - The role of angiotensin II receptors and their antagonists in hypertension. AB - The octapeptide angiotensin II is the major effector of the renin-angiotensin aldosterone system. Angiotensin II causes a variety of potentially noxious biological effects, such as vasoconstriction, a rise in blood pressure, release of aldosterone, enhancement of the effect of catecholamines, and vascular and myocardial hypertrophy, including remodeling of the heart after myocardial infarction. All of these noxious effects of angiotensin II are mediated by angiotensin II receptors (AT receptors) of the AT1 subtype. The functional effects of AT2 receptors, which have been characterized by means of biochemical techniques, are so far not clearly identified. Stimulation of the AT2 receptor by means of angiotensin II is assumed to counteract vascular/myocardial remodeling and possibly to induce vasodilation. Accordingly, AT1 and AT2 receptors are believed to provoke opposite effects. It has drawn attention that fetal tissues contain a high density of AT2 receptors, which is lowered significantly after birth. The identification and analysis of AT receptors has been greatly stimulated by the development of non-peptidergic AT1 receptor antagonists, of which losartan is the prototype. It is so far unclear whether AT receptors are activated in hypertensive disease. A survey will be made of the hemodynamic effects of AT1 receptor antagonists, their interaction with AT receptors, and the probably important role of the sympathetic nervous system involved in the antihypertensive action of AT receptor antagonists. PMID- 10842897 TI - [The role of angiotensin II inhibitors in arterial hypertension: clinical trials and guidelines]. AB - Despite many outstanding favorable results obtained over the past 30-40 years in the treatment of hypertension, several goals of antihypertensive therapy remain unmet. One of them concerns the epidemiological evidence that only 25% of treated hypertensive patients have optimal blood pressure control. Clinical use of new antihypertensive drugs is aimed at achieving the above-mentioned goals, and in particular, at providing more effective blood pressure control, greater cardiovascular protection, and better patient compliance to antihypertensive drug treatment. This paper will examine the main clinical features of angiotensin II receptor blockers, highlighting the results of clinical studies performed thus far, and the main goals of ongoing clinical trials with these drugs. Finally, the role of these compounds in the therapeutic approach to the hypertensive state according to the recent guidelines of the World Health Organization/International Society of Hypertension will be outlined. PMID- 10842898 TI - [Statins: similarities and differences in the pharmacological, clinical and laboratory aspects]. AB - The inhibitors of 3-hydroxy-3-methylglutaryl-CoA reductase (or statins) are the most powerful drugs affecting lipid and lipoprotein levels in plasma. Results obtained from large controlled trials using simvastatin, pravastatin and lovastatin for the primary or secondary prevention of coronary heart disease have demonstrated that treatment with statins is associated with a significant reduction in coronary morbidity and mortality and in total mortality. This is probably due to a more general anti-atherogenic effect of these drugs beyond their lipid-lowering activity. Meta-analysis of data from these large trials indicates that statins have an impact also on the incidence of cerebrovascular events. Currently, six statins have been approved for therapeutic use in different countries. In spite of the similarities in their chemical structure and mechanisms of action, statins may differ in many aspects such as pharmacological properties (hydrophilic vs lipophilic, elimination half-life, cytochrome P450 metabolism, etc.), effects on lipid and other biochemical variables, or pleiotropic effects on different metabolic processes related to atherosclerosis (endothelial function, platelet aggregation, immune function, etc.). In general, the safety and tolerability profile for all statins currently in use is good with a < 2% incidence of undesirable effects. PMID- 10842899 TI - [Atherosclerosis in light of the evidence from large statin trials]. AB - The results of the large trials with the inhibitors of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase (or statins) in terms of clinical benefits in patients with coronary artery disease indicate that these drugs act in a complex way on several pathways involved in the atherosclerotic process. Beyond their lipid-lowering activity, statins appear to modify many characteristics of the arterial wall, resulting in protection against the progression of plaque growth and/or the precipitation of acute events. The so called "pleiotropic" effects of statins, such as the restoration of endothelial activity, the antioxidant potential or the antiproliferative effect on smooth muscle cells, have been investigated mainly in in vitro experiments. In the near future, it will be necessary to unravel the biological significance and the clinical relevance of these effects, which also involve other pathological conditions such as cancer and osteoporosis. Furthermore, lessons from the trials with statins have allowed a better understanding of the mechanisms leading to atherosclerosis which is now considered as an inflammatory process affecting the vascular wall. This has opened new perspectives in the search for the development of newer and more targeted anti-atherogenic drugs. PMID- 10842900 TI - [Atherosclerosis and simvastatin: new questions for the new millennium]. AB - Coronary artery disease is still associated with high morbidity and mortality in Western countries. Lipid blood levels have a tight correlation with the risk of coronary events, and the results of many trials on lipid-lowering therapy (and particularly on simvastatin) demonstrated a significant reduction in total and cardiac mortality, and in the incidence of myocardial infarction and coronary events; even the progression of coronary stenosis has been reduced by treatment with statins. Beyond cholesterol reduction, simvastatin exerts many favorable effects on endothelial function, inflammatory activity, expression of pro thrombotic factors and oxidative stress, yielding a rational basis for its important clinical positive effects, both in primary and secondary prevention of coronary disease. Future developments, which are the subjects of many planned or ongoing clinical trials, are related to the treatment of high-risk patients, the evaluation of the efficacy of elevated simvastatin dosages and of a deep reduction in cholesterol blood levels, the interaction between simvastatin and other drugs (antioxidant compounds, vitamins, antiplatelet drugs) or interventional procedures (percutaneous transluminal coronary angioplasty). Particularly, the Heart Protection Study, the A to Z trial, and the SEARCH and SMART studies will provide important data on a wider, earlier and greater use of simvastatin, which has been demonstrated effective both in the prevention and treatment of acute coronary syndromes. PMID- 10842901 TI - Fatigue, mood, and hemodynamic patterns after myocardial infarction. AB - A descriptive design with repeated measures was used to describe patterns of fatigue, emotional stress, and left ventricular (LV) function among 22 patients with myocardial infarction (MI) from day 5 postadmission to day 21 postadmission for the MI. The severity of fatigue in patients with MI during the subacute period ranged from 32 to 44 on the 100-mm Visual Analogue Scale for Fatigue. Severity of fatigue and depression remained the same; however, LV function improved (p < .01) and patients experienced more energy (p < .01) and less anxiety (p < .01) in the third week following MI. Researchers observed five different fatigue patterns: decreasing fatigue, increasing fatigue, unchanged low fatigue, unchanged-high fatigue, and a curvilinear fatigue pattern. The finding of five different fatigue patterns after an MI suggests that all patients with MI should not be treated as a uniform group assumed to have decreasing fatigue with the passage of time. PMID- 10842902 TI - Communicating pain and pain management needs after surgery. AB - This descriptive study explored how patients communicate their pain and pain management needs after surgery. Thirty postoperative patients were interviewed. The majority described avoiding or delaying communicating their pain at some point during their hospitalization. Reasons for decreased pain communication included not wanting to complain; not wanting to take the provider away from other patients; avoiding unpleasant analgesic side effects; and not wanting to take "drugs." Postoperative patients may be unclear about their role in pain management. Pain management communication problems identified in this study could be used to design intervention studies to improve pain communication and consequent pain relief. PMID- 10842903 TI - Differences in family caregiver outcomes by their level of involvement in discharge planning. AB - Family caregivers play vital roles in assisting elders after they are released from the hospital. Although health care professionals advocate involving family caregivers in discharge planning for elders, little is known about the extent to which this involvement benefits or jeopardizes the caregiver's health and their perceptions of the caregiving experience. The purpose of this study was to determine whether the level of family caregiver involvement in discharge planning for an elder made a difference in caregiver health, discharge planning satisfaction, perception of care continuity, preparedness to assist the elder, and acceptance of the caregiving role 2 weeks and 2 months postdischarge. The sample consisted of 130 family caregivers for elders hospitalized with heart failure. Telephone interviews were conducted 2 weeks and 2 months postdischarge. The findings indicated that family caregivers who reported more involvement in discharge planning had significantly higher scores on satisfaction, feelings of preparedness, and perception of care continuity 2 weeks following the elder's hospitalization than those who reported little or no involvement in planning. Caregivers who reported more involvement in planning also were more accepting of the caregiving role. At 2 months postdischarge, caregivers who reported more involvement in discharge planning reported better health and more acceptance of the caregiving role than those who had little or no involvement in planning. PMID- 10842904 TI - Colonic cleansing, fluid absorption, and discomfort following tap water and soapsuds enemas. AB - Tap water and soapsuds are common enema solutions, but little is known about their effectiveness. Consequently, nurses have no guidance when choosing the best enema for a given patient. In this study, liver transplant patients were given either a tap water or soapsuds enema preoperatively. Amount of enema instilled into the colon and output were measured. Soapsuds enemas produced significantly greater output than tap water and were equally well tolerated. Most subjects who received tap water enemas retained more fluid than was eliminated. Based on these findings, nurses should use caution when giving repeated enemas to patients sensitive to large fluid loads. PMID- 10842905 TI - The role of the pilot study: a case illustration from cardiac nursing research. PMID- 10842906 TI - Measuring behavior: electronic devices in nursing studies. PMID- 10842907 TI - Getting organized: qualitative data collection. AB - A qualitative data collection bag can be an effective tool for maintaining an organized process while collecting qualitative data. Having essential items on hand will facilitate completing data collection in a timely, efficient manner. Timely completion of a project results in positive experiences for both the researcher and participant. PMID- 10842909 TI - Current research initiatives. AB - Patient safety is now in the forefront. Attention to patient safety opens up new opportunities to develop new and better systems for care delivery, and new approaches to ensure patient safety. Research to discover the most effective ways for organizing care delivery and to develop optimal models of nursing care provide the much-needed science on which to base decisions. The AONE Education, Research and Development Priorities provide a framework for this research agenda. The work of cohesive multidisciplinary teams of various professionals would be enhanced by research findings. In reality, there are multiple factors that contribute to error. Seeking depth in understanding of what should be done to reduce error is a matter of professional accountability. PMID- 10842908 TI - An anticoagulation pathway for quality management. PMID- 10842910 TI - Cervical intraepithelial neoplasia--biopsy methods as AIDS for diagnosis and therapy. AB - Between 1977 and 1995, a total of 1610 patients were admitted to hospital with Pap III, IIID, IVa or IVb. The results of 1182 conisations with cervical abrasion and 428 ectocervical surface scrapings with cervical abrasion were evaluated. The percentage of findings of (CIN II was 35.2% for all cases and 23.3% for conisation alone. No atypical findings or CIN I was seen in 13.9% of all conisation cases. The introduction of a second biopsy method (ectocervical surface scraping (ESS) in 1985 resulted in the reduction of cases with a positive histological finding for conisations of (CIN I from 17.7% (1977-1984, n = 468) to 11.3% (1985-1995, n = 714). Evaluation of combined PA and punch biopsy from the ectocervix assisted by colposcopy was performed in 130 cases. Histological correspondence between the methods was seen in 44.6% of the cases. In 22.3% of the patients the highest CIN degree was seen in the biopsy material obtained under colposcopic observation, while in 33.1% of the cases the highest degree of dysplasia was seen in the material obtained by ectocervical surface scraping. Conisation as the sole diagnostic tool can be replaced by combined and simplified biopsy techniques without prejudice to safety. If only PA is performed, the possibility cannot be ruled out that in individual cases an early-stage invasive cervical carcinoma may be overlooked. Conisation performed without evidence of atypical epithelial structures or only CIN I findings must be regarded as over treatment. For all CIN III cases, however, conisation remains the most important form of therapy. PMID- 10842911 TI - [Laparoscopic or laparotomic surgery in the treatment of ovarian benign teratomas?]. AB - The operation treatment of ovarian dermoid cysts by laparoscopy (examined group) and by laparotomy (control group) was compared. The mean size of teratomas measured by USG was similar: in the examined group 59 +/- 20 mm, in the control group 62 +/- 27 mm. There was no significant difference in the operation time: laparoscopy 65 +/- 27 min. (range 35-105 min.) and laparotomy 66 +/- 27 min. (range 40-120 min.). The antibiotic therapy both intra-operation and after operation was applied twice more frequent for laparotomy operated patients. The number of patients with post-operative fever was much higher in the control group (laparotomy: 7 patients--26.9%) than in examined group (laparoscopy: only 1 patient--3.9%). The hospitalization after operation was longer in the control group (mean: 6.8 +/- 3.7 days) than in examined group (mean: 3.1 +/- 2.8 days). Laparoscopic surgery is valuable operating method for selected teratomas in comparison with classical surgery. PMID- 10842912 TI - [The analysis of the coexistence of endometrial carcinoma and uterine myoma]. AB - OBJECTIVE: An epidemiologic study of coexistence uterus myoma with endometrial cancer patients is presented. Coexistence of endometrial carcinoma with uterus myoma was evaluated and controlled for age, residence, civil status, education, parity, menarche age, last menstruation age, length of reproductive period, blood group, hypertension, diabetes, body mass index, sterility, histological subtype, grading, staging. DESIGN: From 1984-1998 136 endometrial carcinomas have been evaluated in the Department of Gynecology & Obstetrics in Hospital of Slupsk retrospectively. MATERIAL AND METHODS: Uterus myoma was coexistence with endometrial cancer in 22 cases (16.2%). Of 136 endometrial carcinomas 16 (11.6%) were multiple malignant neoplasms. Of these neoplasms 9 (6.6%) occur together with breast cancer, 3 (2.2%) with ovarian carcinoma, 1 (0.7%) with stomach carcinoma, 1 (0.7%) with rectum carcinoma, 1 (0.7%) with carcinoma in focus of endometriosis and 1 (0.7%) coexists with double neoplasms (bowel and endometriosis carcinoma) During the 14-year period of study 15 patients (11.0%) out of 136 patients diagnosed as having endometrial cancer had double and 1 (0.6%) had triple primary malignant neoplasms. There was not significant difference in age rate (p = 0.16), residence rate (p = 0.72), civil status rate (p = 0.37), education rate (p = 0.53), parity rate (p = 0.49), menarche age rate (p = 0.33), last menstruation age rate (p = 0.12), length of reproductive period rate (p = 0.66), blood group rate (p = 0.19), hypertension rate (p = 0.38), diabetes rate (p = 0.96), overweight status rate (p = 0.76), sterility rate (p = 0.35), histological subtype rate (p = 0.25), grading rate (p = 0.29), staging rate (p = 0.54), second primary malignant neoplasma (p = 0.77) between both patients group (with and without uterus myoma). CONCLUSION: Patients with endometrial cancer should be carefully and regularly followed up by monitoring et every anatomic site, especially the breast, stomach, and colon, in order that the development of a second primary carcinoma can be detected as early as possible, and not be overlooked in examinations. There is not additional risk factors for endometrial carcinoma between patients with and without coexistence uterus myoma. In the observation of patients with myoma in postmenopause, cytological investigation of endometrial aspirates, ultrasound and mammographic screening should be carried out. PMID- 10842913 TI - [Aromatase (P450AROM) mRNA expression in normal, hyperplastic and malignant endometrium and aromatase activity in endometrial cancer tissue culture]. AB - Aromatase (P450AROM) is the enzyme complex with converts testosterone to estradiol and androstendione to estrone. This enzyme was detected in various normal tissues and uterine pathology such as uterine myoma, endometrial cancer and endometriosis. The aim of the study was to estimate expression of P450AROM messenger ribonucleic acid (mRNA) in normal, hyperplastic and malignant endometrium, and the ability to convert androstenedione to estrone by endometrial cancer tissue. Normal endometrium was obtained from 16 (12 proliferative phase, 4 secretory phase) regularly cycling women after hysterectomy for myomas, hyperplastic endometrium (n = 5) and endometrial cancer (n = 5) from postmenopausal women. The ability to convert androstenedione to estrone was estimated in 16 cases of endometrial cancer in postmenopausal women. P450AROM mRNA was measured by a quantitative assay based on reverse transcribing the mRNA into cDNA with reverse transcriptase (RT) then amplification of the cDNA using the polymerase chain reaction (PCR). The mean (+/- SEM) expression of aromatase gene in proliferative endometrium was 84.4 +/- 14.0 pg mRNA/microgram DNA and in secretory endometrium 200.3 +/- 87.8 pg mRNA/microgram DNA. The mean (+/- SEM) P450AROM mRNA expression in endometrial hyperplasia was 92.9 +/- 17.8 pg mRNA/microgram DNA, in endometrial cancer was 14.3 +/- 7.7 pg mRNA/microgram DNA. Androstenedione to estrone conversion in endometrial cancer tissue culture was 252.5 +/- 91 fmol/g tissue/h. Our data confirm that human normal, hyperplastic and malignant endometrium do express P450AROM mRNA and that aromatase activity is present in endometrial cancer tissue. PMID- 10842914 TI - [Analysis of hemorrhage or delayed postpartum involution of uterus and pathomorphological findings]. AB - OBJECTIVE: The purpose of this study was to analyse the histological findings among woman who underwent curettage during puerperium because of suspecting retained products of conception. MATERIALS AND METHODS: 50 patients staying at the I Clinic of Obstetrics and Gynecology in Warsaw in 1988-1998 because of suspicion of retained products of conception. Study group was divided into two subgroups depending on clinical symptoms such as haemorrhage or delayed postpartum involution of uterus. The first subgroup consisted of 20 woman who underwent curettage during their stay at the hospital between 3 and 11 day after birth. The second subgroup consisted of 30 woman who underwent curettage between 11 and 56 day after birth, they were than readmitted to the hospital. RESULTS: In the first subgroup retained placental fragments were found in 4 patients (20%), decidual tissue in 15 patients (75%) and fragments of myometrium and proliferative endometrium in 1 patient (5%). In the second subgroup, among 30 patients--placental tissue was obtained at curettage in 9 patients (30%), decidual tissue in 13 patients (43.3%), fragments of myometrium and proliferative endometrium were detected in 5 patients (16.7%) and endometritis in 3 patients (10%). CONCLUSION: Haemorrhage or delayed postpartum involution of uterus which is the indication to perform the curettage was in 26% patients the result of retained products of conception. PMID- 10842915 TI - [Urinary N-acetyl-beta-D-glucosaminidase (NAG) activity in women with preeclampsia]. AB - OBJECTIVES: Preeclampsia is the pregnancy specific disorder in which reduced organ perfusion and ischemia of the kidney, liver and brain can be detected. In the kidney, the most typical anatomopathological lesion is glomerular endotheliosis but renal tubular damage was also demonstrated. The NAG is an enzyme which is released into urine from the lysosomes of the proximal renal tubules epithelium and therefore increased urinary NAG activity is a marker of tubular damage or dysfunction. DESIGN: The aim of the study was to assess the renal tubular damage in women with preeclampsia by measuring of urinary NAG activity. MATERIALS AND METHODS: The study group consisted of 46 pregnant women in third trimester with preeclampsia. 36 normal pregnant women in third trimester and 26 healthy nonpregnant women served as two control groups. Urinary activity of NAG was measured in the second morning urine sample by colorimetric method and the results were expressed as NAG/creatinine ratio (U/mmol). RESULTS: Urinary NAG/creatinine ratio was increased in normal pregnant women in third trimester (0.77 +/- 0.44 U/mmol) exceeding that of nonpregnant women (0.23 +/- 0.16). In women with preeclampsia, the urinary NAG/creatinine ratio was significantly higher when compared to the normal pregnant women in third trimester (1.46 +/- 1.63 U/mmol). There were no correlation between urinary NAG excretion and blood pressure results in women with preeclampsia. CONCLUSIONS: The increased urinary NAG activity in women with preeclampsia seems to be a sign of proximal tubular damage. The renal function follow up study in such women is indicated. PMID- 10842917 TI - Hydatidiform mole in 59-years-old woman: a case report. AB - We describe a clinical data and histopathological findings in hydatidiform mole found in 59-years-old woman. The patient was admitted to Hospital with vaginal bleeding. Diagnostic curettage revealed hydatidiform mole VI group according to Hertig-Sheldon classification. The patient underwent transabdominal hysterectomy and bilateral oophorosalpingectomy. The serum level of beta-HCG has decreased from initially observed 128.6 U/l to less than 3 U/l after 3 courses of MTX therapy. Patient is under every-six-month observation. No further treatment is planned. PMID- 10842916 TI - [Elective cesarean section in case of threatened twin pregnancy: a case report]. AB - The authors present the use of computer-aided monitoring of pregnancy in case of twin. In our case one twin was threatened of intrauterine death. The frequency of antepartum death of one twin varies from 0.5%-6.8% of all twin deliveries. The underlying causes for antepartum death are multiple. It suggests that fetal growth retardation is due to competition for uteroplacental circulation and nourishment. In this case, the smaller twin was threatened of intrauterine death, because of intrauterine growth retardation, so the pregnancy was delivered on 32. week of gestational age by cesarean section. PMID- 10842918 TI - [Lipid peroxidation in hypertensive disorders of pregnancy]. AB - The present article discusses potential mechanisms by which lipid peroxidation might contribute to the pathogenesis of hypertensive disorders of pregnancy. PMID- 10842920 TI - Accumulation of diacylglycerol induced by CCl4-derived radicals in rat liver membrane and its inhibition with radical trapping reagent--FT-IR spectroscopic and HPLC chromatographic observations. AB - We have investigated the accumulation of diacylglycerol (DAG) induced by carbon tetrachloride (CCl4)-derived radicals in the liver of female Sprague-Dawley (SD) rats after intraperitoneally injecting CCl4. DAG is an intracellular activator of protein kinase C (PKC) which regulates cell proliferation and differentiation. The electron spin resonance (ESR) study gave the signal of the PBN-CCl3 adduct in the liver of the rats which were pretreated with PBN, confirming that CCl4 was metabolized into CCl3. radicals with cytochrome P450 enzyme and indicating that PBN could trap them. The blood biochemical assay supported the trapping of the CCl3. radicals; the pretreatment of rats with PBN inhibited the increase in the GOT and GPT values upon exposure to CCl4. The Fourier transform-infrared (FT-IR) study indicated in comparison with the model compounds that the CCl4-injected rats accumulated DAG in addition to phosphatidylcholine, phosphatidylethanolamine and triglyceride (TG) in the lipid membrane fraction of the liver homogenate. DAG was found to be ca. 10-15% of the membrane phospholipids by weight. However, DAG was not found in the lipid of the liver microsomes, suggesting that it is formed only in the cell membrane of liver. Also, neither DAG nor TG was found in the lipid membrane of the rats that were pretreated with PBN followed by an injection of CCl4. The formation of DAG was confirmed by an HPLC study. The activation of PKC was observed in liver homogenate in the rats that were injected with CCl4. On the basis of the above findings, it was concluded that the CCl4-derived radicals stimulate PKC through the accumulation of DAG in the liver membrane of the rats. Furthermore, it was shown that PBN has a protective and therapeutic effect against CCl4-induced damage. PMID- 10842919 TI - Postnatal changes in Rho and Rho-related proteins in the mouse brain. AB - To provide information on the role of Rho, a GTP-binding protein, in postnatal development of the brain cells, the change in the levels of Rho protein and Rho related proteins was examined in the brain of mice for two weeks after birth, in parallel with the changes in the activity of marker enzymes for neuronal and glial cells. The activities of acetylcholine esterase and choline acetyltransferase of whole brain homogenate, both of which are neuronal marker enzymes, were progressively increased in an age-dependent manner. The activity of 2',3'-cyclic nucleotide 3'-phosphohydrolase, a glial marker enzyme, increased markedly between one and two weeks after birth. In contrast, the levels of RhoA and RhoB in the membrane fraction were decreased during the postnatal period. The amount of Rho GDP dissociation inhibitor, a regulatory protein for Rho, was unchanged, while those of Rho target proteins, Rock-2 and citron, were gradually increased. Since the inactivation of Rho is known to induce neurite extension and neuronal and glial differentiation in vitro, our results suggest that the Rho signalling pathway plays a regulatory role in the postnatal differentiation of neuronal and glial cells in vivo. PMID- 10842921 TI - Dose dependency of prednisolone on the establishment of Echinococcus multilocularis infection in an alternative definitive host, Mongolian gerbil. AB - This study revealed the dose dependency of prednisolone tertiary-butylacetate (PTBA) treatment on the establishment of Echinococcus multilocularis in the small intestine of Mongolian gerbil (Meriones unguiculatus) and that some of the physiological parameters of host were correlated with the doses of PTBA and establishment of the worm. Twenty Mongolian gerbils were divided into 5 groups, according to the doses of PTBA; 0 mg, 0.5 mg, 2 mg, 5 mg and 10 mg per head. All animals were injected intraperitoneally with PTBA every other day from 6 days before to 6 days after infection. Doses of PTBA and the number of worms recovered at 7 days post-infection showed a positive correlation (r = 0.929, P < 0.0001). The increase of total protein (TP) and the decrease of the percentage of lymphocytes in the peripheral leukocytes were dependent on doses of PTBA (TP: r = 0.811, P < 0.0001, percentage of lymphocyte: r = -0.92, P < 0.0001). The TP and the percentage of lymphocyte also correlated with the number of worms recovered (TP: r = 0.617, P = 0.0049; percentage of lymphocyte: r = -0.800, P < 0.0001). PMID- 10842922 TI - Application of FT-IR and ESR spectroscopic techniques to the study of CCl4 induced peroxidation in rat liver microsomes. AB - FT-Ir and ESR were used for on the investigation of the CCl4-induced peroxidation of rat liver microsomes in combination with biochemical methods. Lipid peroxidation was assayed by TBA reagent in the presence of CCl4 and NADPH. The CCl3. radical was detected by ESR spectroscopy with a spin trapping reagent of PBN. The FT-IR spectroscopy revealed that absorption band of -C-H in -C=C-H decreased in intensity at 3012 cm-1, but the absorption bands of the phosphate head and choline in the phospholipids did not significantly change between 1300 and 900 cm-1. These findings were interpreted to be due to the removal of H. from -C=C-H by radicals as the first step of lipid peroxidation, and to the absence of dephosphorylation of phospholipids in the microsomal membrane. This is the first IR spectroscopic evidence indicating the nature of damage to a microsomal membrane caused by CCl4 treatment. The spectroscopies used here demonstrated that they are useful tools to observe the damage to microsomal membranes. PMID- 10842923 TI - Immunohistochemistry for parathyroid hormone-related protein (PTHrP) in benign and malignant mammary mixed tumors of dogs with and without hypercalcemia. AB - We evaluated the expression of parathyroid hormone-related protein (PTHrP) by immunohistochemistry in eight benign and malignant mammary mixed tumors of dogs with (n = 4) and without (n = 4) hypercalcemia. Positive immunoreactive staining for PTHrP was observed in all four tumors from hypercalcemic dogs. The mammary tumors from 2 of the 4 normocalcemic dogs stained positively for PTHrP, but the numbers of immunoreactive cells and intensity of the immunoreaction were less than in the hypercalcemic dogs. In the other 2 tumors without hypercalcemia, the tissue samples were negative for PTHrP. PMID- 10842924 TI - [Peptic ulcer and gastro-esophageal reflux]. PMID- 10842925 TI - [Pharmaceutical applications of the zeta potential--use in characterization of colloidal drug carriers]. AB - Colloidal drug carriers which mainly involve submicron emulsions, nanoparticles, microparticles, liposomes and lipid complexes have received increasing interest in recent years mainly as vehicles of lipophilic drugs and as improved delivery systems for drug targeting. Size and encapsulation efficiency are, in general, the two parameters used to characterize these pharmaceutical forms. Nevertheless, the surface characteristics of these dispersion have been known to influence their physical, chemical and biological properties. Then, the aim of these study is to evaluate, with some examples and illustrations, the interest of zeta potential determinations to improve the characterization of these colloidal drug carriers. PMID- 10842926 TI - [Reactions and interactions of drugs]. PMID- 10842927 TI - [The carbon dioximeter: a device for quantifying the carbon dioxide released from effervescent pharmaceuticals]. AB - Since effervescent pharmaceuticals are more sensitive to ambient humidity during the manufacturing process and storage, the strict control of their carbon dioxide content becomes a prerequisite to guarantee their physicochemical stability. Indirect gravimetry is a simple and precise method that consists in taking the weight before and after the effervescent reaction allowing to determine the released amount of carbon dioxide. Some authors have used it with devices that lead to longer analysis times and poor accuracy of measurements (due to the excess of released carbon dioxide). The device that we have built (proposed name "CARBONDIOXIMETER") is very easy to set up, and yields quick, accurate, precise and reproducible results. An assay takes three minutes in an acidic medium and five minutes in carbon dioxide free water). Moreover, through the interconnection of scales, registration and treatment of the results can be performed by a printer or a computer. Thus, the "Carbondioximeter" is contributing to the control of physicochemical stability of effervescent pharmaceuticals during the manufacturing process and storage. PMID- 10842928 TI - Effect of stabilizing versus destabilizing interactions on plasminogen activator inhibitor-1. AB - Plasminogen activator inhibitor-1 (PAI-1) is a unique member of the serpin family, as it spontaneously converts into a latent conformation. However, the exact mechanism of this conversion is not known. Previous studies reported that neutralizing monoclonal antibodies (MAs) as well as reversal or removal of charges on the s3C-s4C turn ('gate-region') result in a destabilization of PAI-1 leading to an accelerated conversion to its latent form. In this study the effect of the reversal or removal of charges in this 'gate region' (R186E/R187E, H190E/K191E, H190L/K191L and R356E) on a stable PAI-1-variant (PAI-1-stab) was investigated. PAI-1-stab-R186E-R187E, PAI-1-stab-H190E-K191E, PAI-1-stab-H190L K191L and PAI-1-stab-R356E have a strongly decreased half-life (p < 0.005 versus PAI-1-stab) resulting in half-lives similar or slightly increased to that of wild type PAI-1 (wtPAI-1). Reversal of the positively charged residues at position 186/187 or at position 356 and removal at position 190/191 do not only have a destabilizing effect on the active conformation, but also on the substrate conformation. However, reversal of the charge at position 190/191 does not affect the stability of the substrate conformation. Moreover this study is to the best of our knowledge the first to report that the distal hinge region of the reactive site loop (Arg 356) may be involved in the substrate behaviour of PAI-1. In addition our results suggest an important role for the gate-region for the inactivation of PAI-1 through the conversion to latency both for the destabilizing mutations and for the neutralizing MAs. PMID- 10842929 TI - Synthesis of monodisperse nimesulide nanoparticles in microemulsions E170/isopropyl myristate/water/n-butanol (or isopropanol). AB - Nanoparticles of nimesulide have been synthesized in two systems of microemulsion: E170/isopropyl myristate/water/n-butanol (or isopropanol). Nanoparticles are monodisperse. In the two microemulsions, the size of the nanoparticles is comprised between 45 and 60 A and also seems to be independent of the factor R ([water]/[E170]) and of the concentration of the nimesulide solubilized in chloroform. The constancy of the size suggests that the size is controlled by thermodynamic stabilization of the nanoparticles with the surfactant. The nature of the cosurfactant does not have an obvious influence on the nanoparticle size. The nanoparticles are instantaneously formed and stay stable during a long period of time (several months). PMID- 10842930 TI - Concentration and composition of atmospheric aerosols from the 1995 SEAVS experiment and a review of the closure between chemical and gravimetric measurements. AB - We summarize the results from the various measurements and the inter-sampler comparisons from Southeastern Aerosol and Visibility Study (SEAVS), a study with one of its objectives to test for closure among chemical, gravimetric and optical measurements of atmospheric aerosol particles. Sulfate and organics are the dominant components of the SEAVS fine particles (nominally, particles with aerodynamic diameter < or = 2.5 microns) but between 28 and 42% (range over various samplers) of the gravimetrically measured total fine particle concentration is unidentified by the chemical measurements. Estimates of water associated with inorganic components and measurement imprecision do not totally explain the observed difference between gravimetric and chemical measurements. We examine the theoretical and empirical basis for assumptions commonly made in the published literature to extrapolate total fine particle concentration on the basis of chemical measurements of ions, carbon and elements. We then explore the more general question of closure using the SEAVS data as well as data from other, similar studies reported in the literature. In so combining the SEAVS measurements with other similar studies, we find a strong association between organic carbon and the unidentified component, that is, the fraction of the total fine particle concentration not identified by chemical measurements. We offer several tenable hypotheses for the relationship between the organic and unidentified components that deserve to be tested in future work. Specifically, we hypothesize that (1) errors in the sampling and analysis of organic carbon; (2) estimates of organic mass from measurements of organic carbon; and/or (3) water absorption by organics may all contribute to the observed relationship. PMID- 10842931 TI - Optical measurements of aerosol size distributions in Great Smoky Mountains National Park: dry aerosol characterization. AB - Aerosol size distributions were measured during the summertime 1995 Southeastern Aerosol and Visibility Study (SEAVS) in Great Smoky Mountains National Park using an Active Scattering Aerosol Spectrometer (ASASP-X) optical particle counter. We present an overview of the experimental method, our data inversion technique, timelines of the size distribution parameters, and calculations of dry accumulation mode aerosol density and refractive index. Aerosol size distributions were recorded during daylight hours for aerosol in the size range 0.1 < Dp < 2.5 microns. The particle refractive index used for the data inversion was calculated with the partial molar refractive index approach using 12-hr measured aerosol chemical composition. Aerosol accumulation mode volume concentrations ranging from 1 to 26 micron 3 cm-3 were observed, with an average of 7 +/- 5 micron 3 cm-3. The study average dry accumulation mode geometric volume median diameter was 0.27 +/- 0.03 micron, and the mean geometric standard deviation was 1.45 +/- 0.06. Using an internally mixed aerosol model, and assuming chemical homogeneity across the measured particle distribution, an average accumulation mode dry sulfate ion mass scattering efficiency of 3.8 +/- 0.6 m2 g-1 was calculated. PMID- 10842932 TI - Estimates of particle hygroscopicity during the Southeastern Aerosol and Visibility Study. AB - Aerosol water content was determined from relative humidity controlled optical particle counter (ASASP-X) size distribution measurements made during the Southeastern Aerosol and Visibility Study (SEAVS) in the Great Smoky Mountains National Park during summer 1995. Since the scattering response function of the ASASP-X is sensitive to particle refractive index, a technique for calibrating the ASASP-X for any real refractive index was developed. A new iterative process was employed to calculate water mass concentration and wet refractive index as functions of relative humidity. Experimental water mass concentrations were compared to theoretically predicted values assuming only ammonium sulfate compounds were hygroscopic. These comparisons agreed within experimental uncertainty. Estimates of particle hygroscopicity using a rural aerosol model of refractive index as a function of relative humidity demonstrated no significant differences from those made with daily varying refractive index estimates. Although aerosol size parameters were affected by the assumed chemical composition, forming ratios of these parameters nearly canceled these effects. PMID- 10842933 TI - Light scattering characteristics of aerosols as a function of relative humidity: Part I--A comparison of measured scattering and aerosol concentrations using the theoretical models. AB - The Southeastern Aerosol and Visibility Study (SEAVS) was undertaken to characterize the size-dependent composition, thermodynamic properties, and optical characteristics of the ambient atmospheric particles in the southeastern United States. The field portion of the study was carried out from July 15 to August 25, 1995. As part of the study a relative humidity controlled inlet was built to raise or lower the relative humidity to predetermined levels before the aerosol was passed into an integrating nephelometer or particle-sizing device. Five other integrating nephelometers were operated in various configurations, two of which were fitted with a 2.5 microns inlet. Fine particle (< 2.5 microns) samplers were operated to measure concentrations of sulfate, nitrate, and ammonium ions, organic and elemental carbon, and fine soil. Mass size distributions were measured with an eight-stage, single orifice cascade impactor. Four different strategies for estimating scattering were used. First, an externally mixed model with constant specific scattering coefficients, sulfate ion mass interpreted as ammonium bisulfate, and ammonium bisulfate growth as a function of relative humidity, is assumed. Second, an externally mixed aerosol model, assuming constant dry specific scattering but with sulfate ammoniation and associated composition-dependent hygroscopicity explicitly accounted for, is used. Third, an externally mixed aerosol model, but with sulfate ammoniation, associated growth as a function of relative humidity, and sulfate size distributions, is applied. Fourth, an internally mixed aerosol model with measured sulfur size distributions and estimated size distributions for other species is used with the growth characteristics of the mixture being estimated using the Zdanovskii-Stokes-Robinson (ZSR) assumptions. Only ionic species were considered to be hygroscopic. The second, third, and fourth approaches yield similar results with reconstructed scattering comparing quite favorably with measured scattering. Accounting for sulfate ammoniation and associated water uptake was the most important detail in achieving closure between measurements and modeled scattering. In general, differences between estimated scattering, assuming internally or externally mixed models, was small. These same models were used to estimate wet to dry scattering ratios. The R2 for an ordinary least squares regression between measured and predicted ratios was high (0.71-0.92), and in most cases the scattering ratio was insensitive to modeling assumptions. However, during some sample periods differences between predicted scattering ratios for the different modeling assumptions were as high as 30%. PMID- 10842934 TI - Light scattering characteristics of aerosols at ambient and as a function of relative humidity: Part II--A comparison of measured scattering and aerosol concentrations using statistical models. AB - The eastern United States national parks experience some of the worst visibility conditions in the nation. To study these conditions, the Southeastern Aerosol and Visibility Study (SEAVS) was undertaken to characterize the size-dependent composition, thermodynamic properties, and optical characteristics of the ambient atmospheric particles. It is a cooperative three-year study that is sponsored by the National Park Service and the Electric Power Research Institute and its member utilities. The field portion of the study was carried out from July 15 to August 25, 1995. The study design, instrumental configuration, and estimation of aerosol types from particle measurements is presented in a companion paper. In the companion paper, we compare measurements of scattering at ambient conditions and as functions of relative humidity to theoretical predictions of scattering. In this paper, we make similar comparisons, but using statistical techniques. Statistically derived specific scattering associated with sulfates suggest that a reasonable estimate of sulfate scattering can be arrived at by assuming nominal dry specific scattering and treating the aerosols as an external mixture with ammoniation of sulfate accounted for and by the use of Tang's growth curves to predict water absorption. However, the regressions suggest that the sulfate scattering may be underestimated by about 10%. Regression coefficients on organics, to within the statistical uncertainty of the model, suggest that a reasonable estimate of organic scattering is about 4.0 m2/g. A new analysis technique is presented, which does not rely on comparing measured to model estimates of scattering to evoke an understanding of ambient aerosol growth properties, but rather relies on measurements of scattering as a function of relative humidity to develop actual estimates of f(RH) curves. The estimates of the study average f(RH) curve for sulfates compares favorably with the theoretical f(RH) curve for ammonium bisulfate, which is in turn consistent with the study average sulfate ammoniation corresponding to a molar ratio of NH4/SO4 of approximately one. The f(RH) curve for organics is not significantly different from one, suggesting that organics are weakly to nonhygroscopic. PMID- 10842935 TI - Aerosol light scattering measurements as a function of relative humidity. AB - The hygroscopic nature of atmospheric fine aerosol was investigated at a rural site in the Great Smoky Mountains National Park during July and August 1995. Passing the sample aerosol through an inlet, which housed an array of Perma Pure diffusion dryers, controlled the sample aerosol's relative humidity (RH). After conditioning the aerosol sample in the inlet, the light scattering coefficient and the aerosol size distribution were simultaneously measured. During this study, the conditioned aerosol's humidity ranged between 5% < RH < 95%. Aerosol response curves were produced using the ratio bspw/bspd; where bspw is the scattering coefficient measured at some RH greater than 20% and bspd is the scattering coefficient of the "dry" aerosol. For this work, any sample RH values below 15% were considered dry. Results of this investigation showed that the light scattering ratio increased continuously and smoothly over the entire range of relative humidity. The magnitude of the ratio at a particular RH value, however, varied considerably in time, particularly for RH values greater than approximately 60%. Curves of the scattering coefficient ratios as a function of RH were generated for each day and compared to the average 12-hour chemical composition of the aerosol. This comparison showed that for any particular RH value the ratio was highest during time periods of high sulfate concentrations and lowest during time periods of high soil or high organic carbon concentrations. PMID- 10842937 TI - Exploring spatial patterns of particulate sulfur and OMH from the Project MOHAVE summer intensive regional network using analyses of variance techniques and meteorological parameters as sort determinants. AB - Project MOHAVE (Measurements of Haze and Visual Effects) encompassed a 1-yr field study in the southwestern United States from September 1991 through August 1992. The congressionally mandated study was a joint partnership between the U.S. Environmental Protection Agency, Southern California Edison, and the National Park Service. A major objective of this study was to quantify the potential haze impacts on the nearby Grand Canyon National Park from the 1580 MW coal-fired MOHAVE Power Project (MPP). Any regional impacts from MPP were from secondary fine sulfate. In this paper, we explore the temporal and spatial patterns of particulate sulfur (Sp) and "organic mass by hydrogen" (OMH) during the summer intensive, conducted from mid-July through the end of August 1992. Using an innovative hierarchical pattern recognition classification scheme, we developed 6 groups of Sp and 8 groups of OMH temporally similar behaving patterns in the sampling region. From a regional understanding of synoptic meteorology, these Sp patterns were explainable. We observed two regional gradients. One gradient was a west-to-east decreasing gradient, most likely the result of major sources from urban southern California, including the San Joaquin Valley. The other decreasing gradient was from south-to-north, perhaps the result of emissions emanating from the large urban centers in northern Mexico. The patterns for OMH were not as regionally homogeneous as the patterns for Sp. A west-to-east decreasing gradient was observed for OMH, along with reduced values in the lower Colorado River Valley and some higher values in central and eastern Arizona. The west-to-east decreasing gradient suggests the presence of the Los Angeles urban plume, while the higher values in central and eastern Arizona may be due to biogenic emissions and increased seasonal fires. PMID- 10842936 TI - Use of Project MOHAVE perfluorocarbon tracer data to evaluate source and receptor models. AB - Project MOHAVE was a major monitoring, modeling, and data analysis study whose objectives included the estimation of the contributions of the Mohave Power Project (MPP) and other sources to visibility impairment in the southwestern United States, in particular at Grand Canyon National Park. A major element of Project MOHAVE was the release of perfluorocarbon tracers at MPP and other locations during 50-day summer and 30-day winter intensive study periods. Tracer data (from about 30 locations) were sequestered until several source and receptor models were used to predict tracer concentrations. None of the models was successful in predicting the tracer concentrations; squared correlation coefficients between predicted and measured tracer were all less than 0.2, and most were less than 0.1. PMID- 10842938 TI - Characterization of regional transport and dispersion using Project MOHAVE tracer data. AB - Perfluorocarbon tracers were released continuously from several surface locations and one power plant stack location during the winter (30 days) and summer (50 days) intensive studies as part of Project MOHAVE. Tracers were released in winter from the Mohave Power Plant (MPP) and Dangling Rope, UT, located on the shore of Lake Powell near Page, AZ; and in summer from MPP, the Tehachapi Pass between the Mojave Desert and the Central Valley in California, and El Centro, CA, on the California-Mexico border. At the Tehachapi tracer release site six hour pulses of a separately identifiable perfluorocarbon tracer were released every four days in order to assess the time for the tracer to clear the monitoring network. Daily 24-hr integrated samples were collected at about 30 sites in four states. Limited tracer concentration data with higher time resolution is also available. Graphical displays and analyses identify several regional transport paths, including a convergence zone in the Mojave Desert, the importance of terrain channeling, especially in winter, and a relationship between 24-hr maximum influence function and distance that may prove useful as a scoping tool and to test regional scale air quality models. In winter, Dangling Rope tracer was routinely transported through the entire length of the Grand Canyon, while in summer, MPP tracer was routinely transported over most of Lake Mead. PMID- 10842939 TI - Relating summer ambient particulate sulfur, sulfur dioxide, and light scattering to gaseous tracer emissions from the MOHAVE Power Project. AB - Project MOHAVE was initiated in 1992 to examine the role of emissions from the 1580 MW coal-fired MOHAVE Power Project (MPP) on haze at the Grand Canyon National Park (GCNP), located about 130 km north-north-east of the power plant. Statistical relationships were analyzed between summertime ambient concentrations of a gaseous perfluorocarbon tracer released from MPP and ambient SO2, particulate sulfur, and light scattering to evaluate whether MPP's emissions could be transported to the GCNP and then impact haze levels there. Spatial analyses indicated that particulate sulfur levels were strongly correlated across the monitoring network, regardless of whether the monitoring stations were upwind or downwind of MPP. This indicates that particulate sulfur levels in this region were influenced by distant regional emission sources. A significant particulate sulfur contribution from a point source such as MPP would result in a non-uniform pattern downwind. There was no suggestion of this in the data. Furthermore, correlations between the MPP tracer and ambient particulate sulfur and light scattering at locations in the park were virtually zero for averaging times ranging from 24 hr to 1 hr. Hour-by-hour MPP tracer levels and light scattering were individually examined, and still no positive correlations were detected. Finally, agreement between tracer and particulate sulfur did not improve as a function of meteorological regime, implying that, even during cloudy monsoon days when more rapid conversion of SO2 to particulate sulfur would be expected, there was no evidence for downwind particulate sulfur impacts. Despite the fact that MPP was a large source of SO2 and tracer, neither time series nor correlation analyses were able to detect any meaningful relationship between MPP's SO2 and tracer emission "signals" to particulate sulfur or light scattering. PMID- 10842940 TI - Second generation chemical mass balance source apportionment of sulfur oxides and sulfate at the Grand Canyon during the Project MOHAVE summer intensive. AB - Receptor-based chemical mass balance (CMB) analysis techniques are designed to apportion species that are conserved during pollutant transport using conserved source profiles. The techniques will fail if non-conservative species (or profiles) are not properly accounted for in the CMB model. The straightforward application of the CMB model developed for Project MOHAVE using regional profiles resulted in a significant under-prediction of total sulfate oxides (SOx, SO2 plus fine particulate sulfate) for many samples at Meadview, AZ. In addition, for these samples the concentration of the inert tracer emitted from the MOHAVE Power Project (MPP), ocPDCH, was also under-predicted. A second-generation model has been developed which assumes that separation of particles and SO2 can occur in the MPP plume during nighttime stable plume conditions. This second-generation CMB model accounts for all SOx present at the various receptor sites. In addition, the concentrations of ocPDCH and the presence of other inert tracers of emission from regional sources are accurately predicted. The major source of SOx at Meadview was the MPP, but the major source of sulfate at this site was the Las Vegas urban area. At Hopi Point in the Grand Canyon, the Baja California region (Imperial Valley and northwestern Mexico) was the major source of both SOx and sulfate. PMID- 10842941 TI - Interpretation of trends of PM25 and reconstructed visibility from the IMPROVE network. AB - Under the IMPROVE visibility monitoring network, federal land managers have monitored visibility and fine particle concentrations at 29 Class I area sites (mostly national parks and wilderness areas) and Washington, DC since 1988. This paper evaluates trends in reconstructed visibility and fine particles for the 10th (best visibility days), 50th (average visibility days), and 90th (worst visibility days) percentiles over the nine-year period from 1988-96. Data from these sites provides an indication of regional trends in air quality and visibility resulting from implementation of various emission reduction strategies. PMID- 10842942 TI - An analysis of the yearly changes in sulfur concentrations at various national parks in the United States, 1980-1996. AB - An apparent increasing trend in the summer concentrations of particulate sulfur at Shenandoah (for the time period 1982-1995) and at Great Smoky Mountains (for the time period 1984-1995) has been pointed out by some researchers. Others have suggested that these increasing trends may be an analytical artifact resulting from the switch from the Stacked Filter Units (SFU) measurement system to the IMPROVE (Interagency Monitoring of Protected Visual Environments) measurement system that occurred during the winter of 1987. To obtain a better understanding of the effect of the protocol change, we investigate the changes in the seasonal averages of sulfur concentrations for successive pairs of years for the period 1980-1996 for about 20 national park sites in the United States. For the period 1980-1987, we use sulfur data from the old (SFU) database and for the period 1988 1996, we use the IMPROVE database. Changes from one year to the next similar to that between 1987 and 1988 occurred during other years and seasons suggesting that chance causes alone could perhaps explain it, the degree to which chance could have caused the changes was measured using the permutation test for matched. At the very least, additional information such as side by side readings using SFU and IMPROVE measurement methods, may be needed to better understand any systematic effect in the sulfur measurements that may be ascribable to the protocol change. PMID- 10842943 TI - Trends in the extremes of sulfur concentration distributions. AB - Understanding the response of air quality parameters such as visibility to the implementation of new air quality regulations, population growth and redistribution, and federal land managing practices is essential to the evaluation of air quality management plans on air quality in federal Class I areas. For instance, the reduction of SO2 emissions from large single point sources should result in the decrease of extreme sulfate concentrations, while population growth in geographic areas outside of urban centers could cause a slow widespread increase of sulfate and organic concentrations. The change in federal land managing practice of increased prescribed fire on a year-round basis in lieu of large naturally occurring wild fires could have the same effect; that is, the frequency of high sulfur days increase and low sulfur days decrease as the result of the management practice. Therefore, it is of interest to examine the trends associated with the proportion of days during which the concentration of some aerosol species is above or below a certain threshold and decide whether this proportion of days is increasing or decreasing or shows a lack of trend. This is a direct indication of whether the quality of the environment is improving or worsening, or neither. PMID- 10842944 TI - Evaluation of wind fields used in Grand Canyon Visibility Transport Commission analyses. AB - The Grand Canyon Visibility Transport Commission (GCVTC) was established by the U.S. Congress to assess the potential impacts of projected growth on atmospheric visibility at Grand Canyon National Park and to make recommendations to the U.S. Environmental Protection Agency on what measures could be taken to avoid such adverse impacts. A critical input to the assessment tool used by the commission was three-dimensional model-derived wind fields used to transport the emissions. This paper describes the evaluation of the wind fields used at various stages in the assessment. Wind fields evaluated included those obtained from the Colorado State University Regional Atmospheric Modeling System (RAMS), the National Meteorological Center's Nested Grid Model (NGM), and the National Oceanic and Atmospheric Administration's Atmospheric Transport and Dispersion (ATAD) trajectory model. The model-derived wind fields were evaluated at multiple vertical levels at several locations in the southwestern United States by determining differences between model predicted winds and winds that were measured using radiosonde and radar wind profiler data. Model-derived winds were also evaluated by determining the percent of time that they were within acceptable differences from measured winds. All models had difficulties, generally meeting the acceptable criteria for less than 50% of the predictions. The RAMS model had a persistent bias toward southwesterly winds at the expense of other directions, especially failing to represent channeling by north-south mountain ranges in the lower levels. The NGM model exhibited a substantial bias in the summer months by extending northwesterly winds in the eastern Pacific Ocean well inland, in contrast to the observed southwesterlies at inland locations. The simpler ATAD trajectory model performed somewhat better than the other models, probably because of its use of more upper air sites. The results of the evaluation indicated that these wind fields could not be used to reliably predict source-receptor impacts on a particular day; thus, seasonally averaged impacts were used in the GCVTC assessment. PMID- 10842945 TI - Assessment of the nested grid model estimates for driving regional visibility models in the southwestern United States. AB - The Nested Grid Model (NGM) is a primitive-equation meteorological model that is routinely exercised over North America for forecasting purposes by the National Meteorological Center. While prognostic meteorological models are being increasingly used to drive air quality models, their use in conducting annual simulations requires significant resources. NGM estimates of wind fields and other meteorological variables provide an attractive alternative since they are typically archived and readily available for an entire year. Preliminary evaluation of NGM winds during the summer of 1992 for application to the region surrounding the Grand Canyon National Park showed serious shortcomings. The NGM winds along the borders between California, Arizona and Mexico tend to be northwesterly with a speed of about 6 m/sec, while the observed flow is predominantly southerly at about 2-5 m/sec. The mesoscale effect of a thermal low pressure area over the highly heated Southern California and western Arizona deserts does not appear to be represented by the NGM because of its coarse resolution and the use of sparse observations in that region. Tracer simulations and statistical evaluation against special high resolution observations of winds in the southwest United States clearly demonstrate the northwest bias in NGM winds and its adverse effect on predictions of an air quality model. The "enhanced" NGM winds, in which selected wind observations are incorporated in the NGM winds using a diagnostic meteorological model provide additional confirmation on the primary cause of the northwest bias. This study has demonstrated that in situations where limited resources prevent the use of prognostic meteorological models, previously archived coarse resolution wind fields in which additional observations are incorporated to correct known biases provide an attractive option. PMID- 10842946 TI - Study of the relationship of distant SO2 emissions to Dallas-Fort Worth winter haze. AB - A study was conducted to estimate the changes in wintertime visual air quality in Dallas-Fort Worth (DFW) that might occur due to proposed reductions in SO2 emissions at two steam electric generating plants in eastern Texas, each over 100 km from the city. To provide information for designing subsequent investigations, the haze was characterized broadly during the first year of the study. Meteorological data acquired then demonstrated that, during haze episodes, emissions from only one of the two plants were likely to be transported directly to DFW. Therefore, the second year of the study was centered on just one of the power plants. Air quality was then characterized within the urban area and at rural locations that would be upwind and downwind of the plant during transport to DFW. An instrumented aircraft measured plume dispersion and the air surrounding the plume on selected days. A mathematical model was used to predict the change that would occur in airborne particulate matter concentrations in DFW if SO2 emissions were reduced to reflect the proposed limitations. The contribution of particles in the atmosphere to light extinction was estimated, and simulated photographs were produced to illustrate the visibility changes. The study concluded that the proposed emission reductions would, at most, subtly change perceived wintertime visibility. PMID- 10842947 TI - Modeling of potential power plant plume impacts on Dallas-Fort Worth visibility. AB - During wintertime, haze episodes occur in the Dallas-Ft. Worth (DFW) urban area. Such episodes are characterized by substantial light scattering by particles and relatively low absorption, leading to so-called "white haze." The objective of this work was to assess whether reductions in the emissions of SO2 from specific coal-fired power plants located over 100 km from DFW could lead to a discernible change in the DFW white haze. To that end, the transport, dispersion, deposition, and chemistry of the plume of a major power plant were simulated using a reactive plume model (ROME). The realism of the plume model simulations was tested by comparing model calculations of plume concentrations with aircraft data of SF6 tracer concentrations and ozone concentrations. A second-order closure dispersion algorithm was shown to perform better than a first-order closure algorithm and the empirical Pasquill-Gifford-Turner algorithm. For plume impact assessment, three actual scenarios were simulated, two with clear-sky conditions and one with the presence of fog prior to the haze. The largest amount of sulfate formation was obtained for the fog episode. Therefore, a hypothetical scenario was constructed using the meteorological conditions of the fog episode with input data values adjusted to be more conducive to sulfate formation. The results of the simulations suggest that reductions in the power plant emissions lead to less than proportional reductions in sulfate concentrations in DFW for the fog scenario. Calculations of the associated effects on light scattering using Mie theory suggest that reduction in total (plume + ambient) light extinction of less than 13% would be obtained with a 44% reduction in emissions of SO2 from the modeled power plant. PMID- 10842948 TI - Analysis and simulation of wintertime light scattering by the urban aerosol in Dallas-Fort Worth. AB - Wintertime atmospheric light scattering in Dallas, TX, was estimated through the use of aerosol models. Input data for the aerosol models were provided by measurements of aerosol chemistry, physical particle size distributions, and distributions of particulate sulfur by particle size, and by predictions by an atmospheric simulation model. Light scattering measurements provided a basis for testing the aerosol models. The SCAPE thermodynamic equilibrium model was used to estimate the amount of liquid water associated with particles and the ELSIE Mie scattering model was applied to estimate the resulting light scattering. The calculations were based on aerosol properties measured in Dallas during December 1994 and February 1995, and changes in scattering due to hypothetical changes in the aerosol were predicted. The predicted light scattering was compared to scattering measured by an Optec nephelometer; agreement was within 20% in every case. PMID- 10842949 TI - A preliminary look at source-receptor relationships in the Texas-Mexico border area. AB - Several factors have recently caused visibility impairment at Big Bend National Park, TX, to be of interest. Analyses of historical data collected there have shown that visibility is poorer and fine particle concentrations are higher at Big Bend than at other monitored Class I areas in the western United States. In addition, air masses frequently arrive there after crossing Mexico, where emissions are not well known. During September and October 1996, a field study was undertaken to begin examining the aerosol, visibility, and meteorology on both sides of the border. Results indicate that, during the study, the largest fractions of fine mass and light extinction at Big Bend were due to sulfates and the trace elements most closely associated with sulfate particles were Na and Se. Based on back trajectory modeling and the spatial, temporal, and inter-species relationships in the fine particle concentrations measured during the study, sulfates arrived at the park from both Mexico and the United States. Se was higher in Texas than in Northern Mexico, while V, Pb, Zn, Ni, and Mn were on average much higher in Mexico. PMID- 10842950 TI - Simulation of stack plume opacity. AB - The visual impact of primary particles emitted from stacks is regulated according to stack opacity criteria. In-stack monitoring of the flue gas opacity allows plant operators to ensure that the plant meets U.S. Environmental Protection Agency opacity regulations. However, the emission of condensable gases such as SO3 (that hydrolyzes to H2SO4), HCl, and NH3, which may lead to particle formation after their release from the stack, makes the prediction of stack plume opacity more difficult. We present here a computer simulation model that calculates the opacity due to both primary particles emitted from the stack and secondary particles formed in the atmosphere after the release of condensable gases from the stack. A comprehensive treatment of the plume rise due to buoyancy and momentum is used to calculate the location at which the condensed water plume has evaporated (i.e., where opacity regulations apply). Conversion of H2SO4 to particulate sulfate occurs through nucleation and condensation on primary particles. A thermodynamic aerosol equilibrium model is used to calculate the amount of ammonium, chloride, and water present in the particulate phase with the condensed sulfate. The model calculates the stack plume opacity due to both primary and secondary particles. Examples of model simulations are presented for three scenarios that differ by the emission control equipment installed at the power plant: (1) electrostatic precipitators (ESP), (2) ESP and flue gas desulfurization, and (3) ESP and selective catalytic reduction. The calculated opacity is most sensitive to the primary particulate emissions. For the conditions considered here, SO3 emissions showed only a small effect, except if one assumes that most H2SO4 condenses on primary particles. Condensation of NH4Cl occurs only at high NH3 emission rates (about 25 ppm stack concentration). PMID- 10842951 TI - Examining impacts of visibility and PM strategies before implementation. AB - One of the major challenges facing the world today is defining paths to sustainable futures. Part of the challenge is developing a national energy strategy that promotes an adequate energy supply for the United States, while enhancing environmental quality and maintaining U.S. competitiveness in the world economy. To assist in this challenge, we have developed a screening technique to analyze the effectiveness of different proposed emissions reduction strategies. The technique, referred to as the visibility assessment screening technique (VAST), is designed to examine possible impacts on visibility of emission changes of sulfur oxides, nitrogen oxides, volatile organic compounds (i.e., SO2, NOx, and VOC) and fine and coarse particulate matter (PM). The influence of relative humidity, natural aerosols, and the chemical interconnections among sulfur and nitrogen components of aerosols in determining the effectiveness of Clean Air Act Amendment and other projected energy-related emissions changes on eastern and western visibility are explored. The effectiveness of these strategies on particulate matter impacts and potentially on ozone is also noted. PMID- 10842952 TI - Receptor modeling for elemental source contributions to fine aerosols in New York State. AB - Fine atmospheric particulate material was collected at five sites in upstate New York and analyzed for its trace element composition by instrumental neutron activation analysis. Of the 3700 daily samples collected over a 2-yr period, 1459 were analyzed for 39 elements, providing a large and detailed data set. Factor analysis (FA) was used to identify potential pollution sources or source regions and to construct inorganic source profiles for each. Following FA, the method of absolute factor scores-multiple linear regression was used to estimate the absolute elemental contribution of each of the identified sources. Factor analysis identified nine sources impacting the sampling region. Seven of these were found to be present in varying degrees among of the sampling sites. The other two sources had more localized impacts and were observed at only one of the sites each. Regional sources (such as the midwestern United States and eastern Canada) and crustal/soil material accounted for the greatest amount of the trace elements measured in the collected material. PMID- 10842953 TI - Sampling duration calculations. AB - Routine air quality monitoring produces filter samples that, when analyzed, yield the total amount of the aerosol present in the volume of air drawn by the pump in the monitoring device during the given sampling period. From this we obtain an average concentration of the aerosol for the given duration. The samples are therefore really aggregate samples. A natural question then is "what is the effect of the duration of aggregation on the accuracy and precision of the estimate of the quantity of interest?" The answer depends on a number of factors, such as the quantity that is being estimated: a mean, or an extreme value, or some other quantity; the nature of the measurement error--additive versus multiplicative; the costs of laboratory analyses, and so on. In this paper, we investigate these issues when the interest is in estimating the mean concentration of a specified aerosol species over a fixed time period. In particular, we propose a method for determining a sampling duration that will yield the "best estimate" of the mean concentration for a given cost whenever appropriate statistical assumptions hold. PMID- 10842954 TI - Application of artificial neural networks to modeling and prediction of ambient ozone concentrations. AB - The deterministic modeling of ambient O3 concentrations is difficult because of the complexity of the atmospheric system in terms of the number of chemical species; the availability of accurate, time-resolved emissions data; and the required rate constants. However, other complex systems have been successfully approximated using artificial neural networks (ANNs). In this paper, ANNs are used to model and predict ambient O3 concentrations based on a limited number of measured hydrocarbon species, NOx compounds, temperature, and radiant energy. In order to examine the utility of these approaches, data from the Coastal Oxidant Assessment for Southeast Texas (COAST) program in Houston, TX, have been used. In this study, 53 hydrocarbon compounds, along with O3, nitrogen oxides, and meteorological data were continuously measured during summer 1993. Steady-state ANN models were developed to examine the ability of these models to predict current O3 concentrations from measured VOC and NOx concentrations. To predict the future concentrations of O3, dynamic models were also explored and were used for extraction of chemical information such as reactivity estimations for the VOC species. The steady-state model produced an approximation of O3 data and demonstrated the functional relationship between O3 and VOC-NOx concentrations. The dynamic models were able to the adequately predict the O3 concentration and behavior of VOC-NOx-O3 system a number of hourly intervals into the future. For 3 hr into the future, O3 concentration could be predicted with a root-mean squared error (RMSE) of 8.21 ppb. Extending the models further in time led to an RMSE of 11.46 ppb for 5-hr-ahead values. This prediction capability could be useful in determining when control actions are needed to maintain measured concentrations within acceptable value ranges. PMID- 10842955 TI - Tropospheric sulfate aerosol formation via ion-ion recombination. AB - We propose a source of aerosols in the lower atmosphere associated with the creation, growth, and recombination of ubiquitous cosmogenically generated ions. This particle source should be favorable in the relatively clean, stable marine boundary layer, providing a uniform, continuous fine particle generator in the presence of dimethylsulfide emissions. Through this mechanism, new sulfate aerosols can be formed at sulfuric acid vapor partial pressures well below the supersaturations required for homogeneous binary nucleation of sulfuric acid/water solutions, which is consistent with numerous observations of new particle formation under sub-saturated conditions. The evolving aerosols in turn control the acid vapor concentration and thus modulate the sizes of the precursor ions and the rate of new particle formation. A simple model representing this nonlinear coupled system predicts that the physical and chemical processes connecting ions, vapors, and aerosols effectively constrain the particle population to a relatively narrow range of values. This self-limiting behavior may explain in part the apparent stability of the marine sulfate aerosol, with mean concentrations of the order of several hundred per cubic centimeter. PMID- 10842956 TI - [Video-laparoscopic suture of duodenal ulcers perforations]. AB - The authors analyze their experience of 62 video-laparoscopic operations for perforating duodenal ulcer from January 1998 to May 1999. The optimal algorithm of the patients' preoperative examination and substantiation of the used technique are given. Basic steps and technical aspects of the operation are designated. Basic approaches to the postoperative anti-ulcerous treatment are specified. Basic indications and contraindications to video-laparoscopic-closure of duodenum opening in perforating ulcer are indicated. PMID- 10842957 TI - [Choice of cholecystolithiasis treatment]. AB - Since January 1996 to December 1998 in the surgical department of Central Clinical Hospital (CCH) 594 cholecystectomies were performed. Laparoscopic cholecystectomies were made in 516 (86.8%) patients. Intraoperative complications occurred in 8 patients, without lethal outcomes. In the gastroenterological department of CCH 146 patients had cholelithic therapy with mechanical lithotripsy. It was possible to completely eliminate fragments of stones in 45 patients. The rest (101 patients) have undergone cholecystectomy. Cholelithic therapy with mechanical lithotripsy is indicated only in young patients it they reject cholecystectomy. PMID- 10842958 TI - [Functional condition of gallbladder after stomach resection by Roux]. AB - This study examined the results of surgical treatment of 90 patients with ulcerative stenosing disease of the stomach and duodenal ulcer between 1984 and 1995. 30 patients (study group) underwent stomach Roux resection. Truncal vagotomy with stomach Bilroth-I resection (control group) was made in 20 patients, 20 patients had a truncal vagotomy with pyloroplasty according to Heineke-Mikulicz (control group), and 20 patients had a selective proximal vagotomy with gastroduodenostomy by Joboulay (control group). Motor and evacuation functions of gallbladder were assessed by dynamic US and radioisotope scintigraphy. After a Roux stomach resection and a stomach Bilroth-I resection, respectively, hypokinetic and hyperkinetic types of the gallbladder's dyskinesia was established. After a selective proximal vagotomy with gastroduodenostomy by Joboulay and truncal vagotomy with pyloroplasty according to Heineke-Mikulicz essential change of the gallbladder refractive function wasn't observed. PMID- 10842959 TI - [Treatment of malignant pancreatic islet cell tumors]. AB - Among 142 patients with islet cell tumor who were admitted to hospital between 1982 and 1999, 15 patients (10.6%) had malignant tumours. The mean age was 45.3 +/- 5.8 years. Islet cell tumors were located in the head and corpus of the pancreas in 26.6% and 26.6% of the patients, respectively, in the tail of the pancreas in 46.8% of the patients. The mean size of the tumor was 2.9 +/- 0.7 cm, in 6 patients (40%) liver metastases were found. US, CT and angiography which sensitivity were 72.7, 100 and 85.7%, respectively, were used for topical diagnosis of the islet cell tumors. 12 patients were operated on (15 operations) and 3 patients were treated conservatively by streptozocin. Surgical procedures included distal pancreatic resection (n = 11), enucleation of the tumor (n = 2), hepatic resection (n = 1), abdominal exploration (n = 1). In 2 patients palliative pancreatic resections were combined with transarterial embolization and devascularisation of the liver metastases and 2 patients were treated by systemic chemotherapy. To reveal malignancy before and during the operation was possible only by evaluation of distant metastases and the adjacent organs infiltration. In other cases to determine the diagnosis was possible only after planned histological identification of the resected specimens. 5-year survival rates after conservative and surgical treatment were 33% and 62.5%, respectively. In patients without distant metastases 5-year survival rate was 100%. PMID- 10842960 TI - [Laparoscopy in diagnosis of intestinal mesentery acute circulatory disturbance]. AB - Laparoscopic data on 74 patients with jejuno-ileal mesentery acute circulatory disturbance were analysed. Positive (diagnosis is confirmed) and negative (diagnosis is rejected) results of the investigation, reasons of false diagnosis and repeated laparoscopies were examined. Features of the jejuno-ileal mesentery acute circulatory disturbance laparoscopic picture are determined depending on its stage (ischemia, infarct, peritxnitis) and kinds (arterial thrombosis and embolism, venous thrombosis, unocclusive mesenteric ischemia). PMID- 10842961 TI - [Contemporary trends in surgery of adrenal glands]. AB - There have been analysed the records of suprarenectomy in 459 patients with Conn' syndrome, syndrome of Icenko-Cushing as well as with androsteroma, pheochromocytoma and other tumors, who were operated in the clinic of abdominal surgery of S.P. Fedorov Russian Military Medical Academy. On the base of retrospective analysis the most actual aspects in surgery of the adrenal glands and the ways of the development of this branch of medicine have been determined. High effectiveness and perspectiveness of endovideosurgical removal of the adrenal glands is shown. Original and tested methods of choice of surgical approach, prophylaxis and treatment for some complications of suprarenectomy (adrenal insufficiency, Nelson' syndrome, etc.) are proposed. Substantiated policy in so-called hormono-inactive tumors of the adrenal glands is proposed. PMID- 10842962 TI - [Treatment of hydatid cysts' residual cavities (experimental and clinical study)]. AB - Experimental series of 60 cotton rats and 150 white mice infected with echinococcus showed that 80-100% glycerin and 3% peroxide solution were effective scolicidal solutions for inactivation of the parasite. 344 patients have been operated on for hydatid disease with inactivation of the parasite by means of these scolicidal solutions. Glycerin during the surgical management of the hydatid cysts is effective and harmless in uncomplicated hydatid liver disease and any hydatid pulmonary cysts. Glycerin is contraindicated in complicated hydatid liver disease because of immunity disturbance. In complicated hydatid liver disease management of the hydatid cysts by 3% peroxide is effective and harmless. It is necessary to correct the hydatid liver disease patients' immunity in the preoperative and postoperative periods. Tactivin and antiechinococcus drug SK-1 are effective immunomodulators in hydatid liver disease. PMID- 10842963 TI - [Surgical treatment of long bones' chronic osteomyelitis]. AB - The experience in surgical treatment of 269 patients with long bone chronic osteomyelitis based on the principles of active surgical policy for purulent wounds is summarized. Some aspects of osteomyelitis surgical treatment are reconsidered and complemented. Variants of osteomyelitic focus surgical debridement dependent on severity the lesion as well as methods of plastic replacement of bones' and soft tissues' defects are demonstrated. PMID- 10842964 TI - [Role of midocalm in combined treatment of neuropathic diabetic foot syndrome]. PMID- 10842965 TI - [Penetrating neck injury by a sword]. PMID- 10842966 TI - [Isolated splenic tuberculosis]. PMID- 10842968 TI - [Acute destructive pyelonephritis and carbuncle of right kidney as a rare complication of gangrenous perforative appendicitis]. PMID- 10842967 TI - [New approaches to autodermoplastic reconstruction of ventral hernias]. PMID- 10842969 TI - [Rare forms of intestinal obstruction]. PMID- 10842970 TI - [Possibilities of diffuse pulmonary emphysema surgical treatment]. PMID- 10842971 TI - [Duodenal injuries]. PMID- 10842972 TI - [Azygos-portal veins disconnection for gastroesophageal varices (review of international literature)]. PMID- 10842973 TI - [Wounding in a duel of famous Russian poet A.S.Pushkin]. PMID- 10842974 TI - DocLink alerts reduce errors. PMID- 10842975 TI - National registry of clinical trials. PMID- 10842976 TI - Medicine meets virtual reality 2000. PMID- 10842977 TI - Computers, grandmasters, and clinical decision making. PMID- 10842978 TI - Evaluating clinical information systems: a step towards reducing medical errors. PMID- 10842979 TI - Metadata and the reintegration of clinical information: ISO 11179. PMID- 10842980 TI - The granting game: Part I. A guide to NLM funding for informatics research. PMID- 10842981 TI - Data mining and modeling: power tools for physician practices. PMID- 10842982 TI - Visionary informatics: the University of Missouri-Columbia Health Sciences Center. PMID- 10842983 TI - Case management. Making the case for information systems. PMID- 10842984 TI - E-pharmacy systems. Prescription and medication fulfillment come of age. PMID- 10842985 TI - International efforts in informatics. Creating a global village for healthcare. PMID- 10842986 TI - HIPAA update. Achieving compliance with the new standards. PMID- 10842987 TI - Integrating the healthcare enterprise. PMID- 10842988 TI - Malpractice affidavit of merit has teeth. PMID- 10842989 TI - . . . Even medical liability insurance. PMID- 10842990 TI - Live and then give. The facts about organ donation. PMID- 10842991 TI - Physicians for responsible negotiation: launching a non-union. PMID- 10842992 TI - [Thwarted genius: the life of Giuseppina Cattani (dedicated to colleagues, also unrecognized]. PMID- 10842993 TI - [Breast cancer and lymph node metastasis: relation of lymph node dimensions and metastasis]. AB - The Authors have examined 1159 lymph nodes obtained from 64 cases of breast carcinoma to value the presence of a correlation between the insurgence of metastases and lymph node dimensions. Metastases were more frequent in larger lymph nodes. Nevertheless, metastases were also present in small (< 5 mm) or very small (< 2 mm) lymph nodes. This indicates the importance to carefully examine small lymph nodes during gross examination and to obtain multiple sections from them. PMID- 10842994 TI - [Creating low-power photomicrographs by digital scanning of histological sections]. AB - The recent introduction of digital film scanners in the medical-scientific field provided everyone dealing with light-microscopy an easy method to obtain low power photomicrographs from entire histological sections. Direct scanning of the common microscope slide allows the easy creation of high quality images. The digital images obtained may be improved or retouched using dedicated software and then printed on paper or film. The required instrumentation is relatively unexpensive and neither skilled staff nor expertise of photographic techniques is required. This reduces costs and saves time. The obtained images may be employed in histopathology, embryology, histochemistry, image analysis and telepathology, and are suitable for scientific papers, educational purposes and lecturing, as well as for daily reporting. The authors bring their personal experience in this field, making also a complete literature review on the topic. PMID- 10842995 TI - [Accessory parameters in the histopthology of hepatitis C. Comparative analysis]. AB - Fifty three consecutive cases of chronic hepatitis C ("b" series) were considered and strictly (numerically and qualitatively) compared with the series ("a") reported by Scheuer et al. Two groups (gr.) respectively inclusive of chronic persistent hepatitis, chronic lobular hepatitis and mild/moderate chronic active hepatitis (gr. I) and severe chronic active hepatitis, developing cirrhosis and established cirrhosis (gr. II) were identified, studied and scored according to the following accessory histological parameters: "lymphoid aggregates and follicles" (LA&F), "bile duct damage" (BDD), "acidophil bodies" (AB), "lymphocytic infiltration of sinusoids" (LIS) and "fatty changes" (FC). In order to interpret the results the "b" series was further examined in relation to lobular "spotty necrosis" (SN) and subtle morphology of FC (type of vacuolarity and topography). Moreover dysplasia (in "a" and "b") and other minor accessory parameters (in "b" only) were briefly considered. In conclusion the comparison of "b" versus "a" confirms a characteristic histomorphological pattern in chronic hepatitis C. As far as the major accessory parameters are concerned the reproducibility was acceptable for BDD and FC, but inadequate for LA&F, LIS and AB. In particular the following points seem to be the most critical challenges: reassessment of the morphological criteria for LA&F, LIS and AB; improvement of the analysis of FC; elucidation of the relation between LIS, AB, SN and LA&F; ascertainment of sensibility and specificity of the accessory parameters in relation to "grading", "staging", differential diagnosis, post-therapy evaluation and non histological findings. PMID- 10842996 TI - [Quality control in pathology: prospective peer review on routine cases]. AB - To describe a prospective method of quality assurance adjusted to an ideal group of pathologists, easily feasible in routine surgical pathology. The Authors examined the same slides (EE) independently and without time-limits. In the same session the individual diagnosis were compared and discussed in order to provide only one conclusive diagnoses. After the discussion, diagnostic agreement is 100%. Peer and simultaneous review yields sufficient benefits in increasing the accuracy of diagnostic reports. It stimulates discussion of diagnostic criteria and terminology, creates more uniform reports, and serves as an educational vehicle for pathologists. PMID- 10842997 TI - [Atypical meningioma: morphometric analysis of nuclear pleomorphism]. AB - The revised edition of the WHO classification of brain tumours now includes the "atypical" meningioma (grade II) which should be placed between the common type (grade I) and anaplastic type (grade III) according to histomorphological features and prognosis. However, diagnostic criteria for atypical meningioma are vague and the significance of brain invasion in the determination of malignancy is controversial. Nuclear pleomorphism and mitoses are usually considered the most important parameters to distinguish atypical and malignant meningiomas. According to WHO classification we selected eight cases of meningioma diagnosed as atypical (3 cases) and malignant (5 cases). All the tumours were supratentorially located. Nine cases of benign meningiomas were also studied as a control group. Morphometrical analysis was carried out by S.A.M. (Shape Analytical Morphometry) system. S.A.M. logical architecture assumes that each irregular shape contains elements of two distinct logical domains: gross distortions that interest the contour and its local perturbations. These features were investigated separately by analytical procedures to acquire independent parameters both on the logical and the numerical level. The results, statistically evaluated, show that nuclear pleomorphism is not a satisfactory criterion, if used alone, to distinguish atypical from malignant meningioma (Discriminant Analysis: 19% of minimum error). PMID- 10842998 TI - [Gastric antral vascular ectasia (GAVE or watermelon stomach): a case report]. AB - Gastric antral vascular ectasia (GAVE), recently described in literature, is an important cause of gastric bleeding. A case of great gastrectasia with vascular ectasia in an elderly male patient is reported. The lesion was characterized by great linear and brush folds along the antrum and body of the stomach, Endoscopically, this folds seems like the watermelon streakings. It could be an acquired lesion probably linked to traumatism on gastric mucosa. PMID- 10842999 TI - [Digital pacinian hyperplasia. Report of a case associated with foreign body reaction]. AB - Digital Pacinian Hyperplasia is a non tumoral, very rare lesion. To date, only 31 cases have been reported, occasionally named Digital Pacinian Neuroma. Digital pain is usually associated to. Local trauma is reported in about half of the cases. We report a new case occurred in a 62 year-old male, associated with inflammatory and foreign body reaction. No recurrence was observed 6 months after surgical excision. Differential diagnosis and controversies about this lesion are discussed. PMID- 10843000 TI - Classification vs. identification in diagnostic quantitative pathology. PMID- 10843001 TI - [Guidelines for the reporting of skin melanocytic lesions. Study Group "Reporting of skin melanocytic lesions" Italian Association of Dermatopathology (IAD)]. PMID- 10843002 TI - [Johann Wolfgang Goethe (1749-1832): creator of the term and concept of morphology]. PMID- 10843003 TI - [A dental substance-sparing procedure for esthetic care of devitalized anterior teeth]. PMID- 10843004 TI - [Sterilization in dental practice: definitions, procedures, European norms and recommendations (III). The Commission for Practice Hygiene, Societe Suisse d'Odonto-Stomatologie]. PMID- 10843005 TI - Responding to the responders. PMID- 10843006 TI - Junk the system. PMID- 10843007 TI - TSBME regulation needed. PMID- 10843008 TI - You can do more to prevent and treat stroke. PMID- 10843009 TI - MedBytes. PMID- 10843010 TI - HMO liability. PMID- 10843011 TI - Antitrust relief. PMID- 10843012 TI - Expanding the frontiers. PMID- 10843013 TI - The science of success. PMID- 10843014 TI - Following through. PMID- 10843015 TI - Prophylaxis against respiratory syncytial virus in high-risk infants: administration of immune globulin and epidemiological surveillance of infection. AB - Hospitalizations secondary to respiratory syncytial virus (RSV) infection in children were monitored before and after the administration of RSV immune globulin. Monitored also was the effectiveness of prophylaxis in preventing the disease among at-risk children along the Texas-Mexico border. Small premature infants were at the greatest risk of hospitalization; preterm infants and then term infants followed. Prophylaxis does not necessarily prevent RSV bronchiolitis but may reduce the severity of the pulmonary disease. PMID- 10843016 TI - Fen-phen manufacturers, not physicians, are liable. PMID- 10843017 TI - Practicing medicine on Navajo reservation proves eye-opening and rewarding. PMID- 10843018 TI - Who manages care? Counterpoint. PMID- 10843019 TI - A work in progress. PMID- 10843020 TI - Wisconsin's children need more calcium. PMID- 10843021 TI - Teens today facing familiar problems. PMID- 10843022 TI - In doc we trust. Building rapport with young patients takes time and skill. PMID- 10843023 TI - Newborn screening in Wisconsin: program overview and test addition. AB - The prevention of congenital anomalies and their sequelae is an important public health objective. One strategy for preventing morbidity and mortality due to congenital disorders is Wisconsin's Newborn Screening Program. Wisconsin has been a national leader in newborn screening since its inception with phenylketonuria screening in 1966. Wisconsin's program is a collaborative effort of the State Maternal and Child Health Program, housed within the Division of Public Health of the Department of Health and Family Services; the State Laboratory of Hygiene; physicians and other health professionals; and families. After in-depth consideration, the Department recently approved the addition of 14 fatty acid oxidation disorders and organic acidemias. This paper provides an overview of the Newborn Screening Program and introduces the disorders that Wisconsin added to the newborn screening panel in April 2000. Technologic advances have provided Wisconsin with yet another tool to improve the health of its citizens and prevent significant morbidity and mortality. PMID- 10843024 TI - Newborn hearing testing at Wisconsin hospitals: a review of the need for universal screening. AB - Every year, an estimated 200 babies are born in Wisconsin with hearing impairment. Early identification of hearing loss is associated with better language development. This study describes the prevalence of newborn hearing screening at Wisconsin hospitals. Ninety-six percent of all 107 acute care hospitals with newborn delivery services responded to a mailed survey. In 1997, 10% of newborns in Wisconsin had hearing tested. In 1997 and 1998, two and then seven hospitals, respectively, established universal hearing screening programs. Nurses performed hearing tests on most newborns using automated auditory brainstem response equipment. Universal newborn hearing screening can be done by nurses or other personnel at community hospitals prior to discharge. PMID- 10843025 TI - Evaluation of a pregnancy outcome risk reduction program in a local health department. AB - This study assesses the effectiveness of the Waukesha County Public Health Department's Prenatal Care Coordination Program (PNCC) in reducing self-reported risk factors for adverse pregnancy outcomes in pregnant clients, through assessment, preventive counseling and follow-up. There were 166 clients seen in the program. They were seen at up to three follow-up visits, with 83 seen at the 12-week follow-up visit, 106 seen at the 24-week follow-up visit, 74 seen at the 38-week follow-up visit, and 151 seen for at least one follow-up visit. Statistically significant improvement was seen between the first and last follow up visit for each of the five categories of risk factors that were assessed. Mean alcohol intake decreased from 9.6 drinks per month at baseline to none by the 38 week follow-up visit. Mean cigarette smoking decreased from 12.8 cigarettes per day to 2.4. Street drug use decreased almost completely. There were statistically significant increases in milk, meat, fruit, bread, and vegetable consumption, and folic acid and multi-vitamin supplementation. PMID- 10843026 TI - Racial disparities in African American and white infant mortality United States and Wisconsin, 1980 to 1998. AB - Infant mortality is a powerful indicator for assessing the health of a population, and the extent to which society invests in its children. This paper examines infant mortality trends in the United States and Wisconsin among the African American and white populations from 1980 to 1998. Data from the National Center for Health Statistics and Wisconsin Birth and Infant Death Reports were used to examine overall infant mortality trends from 1980 to 1998 for African American and white infants. The overall infant mortality rate in Wisconsin decreased steadily from 10.3 infant deaths per 1000 live births in 1980 to 7.2 in 1998. White infant mortality followed this trend, declining from 9.6 in 1980 to 5.6 in 1998. African American infant mortality has remained about the same since 1980 at 18 infant deaths per 1000 live births, even though the overall percentage of African American live births in Wisconsin increased from 6% in 1980 to 10% in 1998. The ratio of African American to white infant mortality rates in Wisconsin increased from 2 to 1 in 1980 to 3.2 to 1 in 1998. While the Wisconsin African American infant mortality rate remained even since 1980, the US rate declined from 22.2 in 1980 to 14.1 in 1998. The Wisconsin African American rate in 1998, 17.9, surpassed the national rate. The use of 5-year running averages to smooth out year-to-year fluctuations showed statistically significant declines in Wisconsin white infant mortality rates and no change in Wisconsin African American infant mortality rates. These trends show the importance for Wisconsin to sustain its current efforts to reduce black infant mortality. At the same time, Wisconsin-based research efforts to determine strategies and factors that work should continue and influence the future design of systems, programs, and policies to eliminate the disparity. PMID- 10843027 TI - Benefits of breastfeeding: a review for physicians. AB - Physicians can support breastfeeding for their patients by providing prenatal families with accurate and current information on the benefits of breastfeeding, eliminating barriers to breastfeeding in both their office and hospital environments, and recognizing the significant impact the physician may have on the breastfeeding practices of their patients and the support for breastfeeding in their communities. PMID- 10843028 TI - The need for a second dose of ceftriaxone in febrile infants age 4-8 weeks. AB - OBJECTIVE: To determine if a second intramuscular injection of ceftriaxone was necessary in febrile infants who meet low-risk criteria for outpatient therapy. SETTING: Children's Hospital Emergency Department. PATIENTS: Febrile infants 4-8 weeks of age. METHODS: Outpatient treatment criteria included non-toxic appearance, no identifiable source for infection on physical examination, CSF WBC > or = 10/mm3, peripheral WBC < or = 15,000/mm3, normal UA, and normal chest radiograph study, if obtained. Additional requirements included a reliable caretaker and re-evaluation in 24 hours. Infants who met these criteria received intramuscular ceftriaxone 50 mg/kg with instructions to follow-up in 24 hours. At the follow-up visit, infants with no identifiable source for infection and negative cultures received a second dose of intramuscular ceftriaxone and were discharged. Cultures were read at 48 hours and at the conclusion of the study. Medical records were reviewed to identify delayed complications. RESULTS: 172 infants were enrolled. The mean age was 45 days. All CBC, UA, CSF analyses were normal. Chest radiographs were obtained in 56 (30%) infants; all were normal. One (.05%) patient was admitted at the follow-up visit. The mean time to follow-up was 25 hours. Two positive cultures were identified at the re-evaluation visit; one blood culture grew Salmonella and a urine culture grew E. coli. The CSF cultures were all negative at follow-up and remained negative. CONCLUSION: Febrile infants 4-8 weeks of age who meet outpatient therapy criteria and have negative cultures and no identifiable source for infection at 24 hours may not require the second dose of intramuscular ceftriaxone. PMID- 10843029 TI - Trends in smoking during pregnancy, Wisconsin 1990 to 1996. AB - PURPOSE: Smoking during pregnancy can be harmful to the fetus. This paper examines trends in smoking during pregnancy in the United States and Wisconsin from 1990-1996. In addition, percentage change in smoking prevalence during pregnancy by selected maternal characteristics and in all Wisconsin counties is compared for the periods 1990-1992 and 1994-1996. METHODS: Data from the National Center for Health Statistics and Wisconsin resident birth certificates were compared to examine overall smoking trends for pregnant women from 1990-1996. For Wisconsin, 3-year averages (1990-1992 and 1994-1996) were calculated to provided more stable estimates. The percentage change in smoking prevalence was examined by selected maternal characteristics, such as age, marital status, race, prenatal care and education, and by county of residence. RESULTS: Smoking rates for pregnant women in the United States have gradually decreased, from 18.4% in 1990 to 13.6% in 1996. From 1990 to 1996, maternal smoking in Wisconsin declined from 22.9% to 18.2% (significant at P < .001). In Wisconsin, smoking rates during pregnancy were higher in both 1990-1992 and 1994-1996 among women who were younger, unmarried, American Indian, or African American; received late or no prenatal care and did not graduate from high school. Out of 72 Wisconsin counties, 63 showed a decrease in smoking rates for pregnant women. CONCLUSIONS: Smoking prevalence during pregnancy in Wisconsin is above the Healthy People 2000 and Wisconsin Public Health Agenda for the Year 2000 goals of less than 10% of women smoking during pregnancy. Public health must continue its initiative to encourage healthy life styles for pregnant women. PMID- 10843030 TI - Helping Wisconsin women quit smoking: a successful collaboration. AB - The cost of treatments for tobacco dependence frequently presents a financial barrier to their use. To overcome such barriers, the Wisconsin Women's Health Foundation, the Wisconsin Bureau of Public Health, the McNeil Consumer Healthcare, and the University of Wisconsin Center for Tobacco Research and Intervention collaborated in an initiative to distribute nicotine patches to Wisconsin women at no cost. As a result of this collaborative effort, approximately 19,000 women received a 6-week course of Nicotrol Patches. To evaluate the effectiveness of this initiative, a sample of 500 recipients were contacted and surveyed by telephone 6 months after receiving their patches. Approximately 22% of these women reported total abstinence at 6 months, and another 77% reported they had reduced their smoking. At follow-up, women who had successfully quit rated their health status significantly better than women who were still smoking. More than 99% of respondents recommended that the program be repeated. Extrapolating the observed abstinence rate to the 19,000 patch recipients, an estimated 4000 Wisconsin women successfully quit smoking as a result of this program. PMID- 10843031 TI - Validation of publication of new names and new combinations previously effectively published outside the IJSEM. PMID- 10843032 TI - Notification that new names and new combinations have appeared in volume 50, part 2, of the IJSEM. PMID- 10843033 TI - Methylocella palustris gen. nov., sp. nov., a new methane-oxidizing acidophilic bacterium from peat bogs, representing a novel subtype of serine-pathway methanotrophs. AB - A new genus, Methylocella, and a new species, Methylocella palustris, are proposed for three strains of methane-oxidizing bacteria isolated from acidic Sphagnum peat bogs. These bacteria are aerobic, Gram-negative, colourless, non motile, straight and curved rods that utilize the serine pathway for carbon assimilation, multiply by normal cell division and contain intracellular poly beta-hydroxybutyrate granules (one at each pole). These strains use methane and methanol as sole sources of carbon and energy and are moderately acidophilic organisms with growth between pH 4.5 and pH 7.0, the optimum being at pH 5.0-5.5. The temperature range for growth is 10-28 degrees C with the optimum at 15-20 degrees C. The intracytoplasmic membrane system is different from those of type I and II methanotrophs. Cells contain an extensive periplasmic space and a vesicular membrane system connected to the cytoplasmic membrane. The strains grew only on media with a low salt content (0.2-0.5 g l(-1)). All three strains were found to possess soluble methane monooxygenase and are able to fix atmospheric nitrogen via an oxygen-sensitive nitrogenase. No products were observed in a PCR with particulate methane monooxygenase-targeted primers; hybridization with a pmoA probe was also negative. The major phospholipid fatty acids are 18:1 acids. The G+C content of the DNA is 61.2 mol%. The three strains share identical 16S rRNA gene sequences and represent a novel lineage of methane-oxidizing bacteria within the alpha-subclass of the class Proteobacteria and are only moderately related to type II methanotrophs of the Methylocystis-Methylosinus group. The three strains are most closely related to the acidophilic heterotrophic bacterium Beijerinckia indica subsp. indica (96.5% 16S rDNA sequence similarity). Collectively, these strains comprise a new species and genus Methylocella palustris gen. nov., sp. nov.; strain KT (= ATCC 700799T) is the type strain. PMID- 10843034 TI - Assignment of Eubacterium sp. VPI 12708 and related strains with high bile acid 7alpha-dehydroxylating activity to Clostridium scindens and proposal of Clostridium hylemonae sp. nov., isolated from human faeces. AB - Unknown Eubacterium-like organisms VPI 12708 and five strains (Y-1113, I-10, M 18, TH-82 and 36S) had high bile acid 7alpha-dehydroxylating activity; the unknown Clostridium-like organisms TN-271T and TN-272 also had the same activity. Analysis of their 16S rDNA sequences demonstrated that all strains belong to cluster XIVa of the genus Clostridium (Collins et al., 1994). Strain VPI 12708 and five other strains (Y-1113, I-10, M-18, TH-82 and 36S) formed a single cluster and strains TN-271T and TN-272 formed another single cluster. Clostridium scindens JCM 6567T was the most closely related species for two clusters in the phylogenetic tree. Values for DNA-DNA similarities among C. scindens JCM 6567T, strain VPI 12708 and the other five strains were greater than 70%, showing that these micro-organisms were a single species. Therefore, we identified strain VPI 12708 and the five other strains as C. scindens. In addition, DNA-DNA similarities among C. scindens JCM 6567T, strain TN-271T and strain N-272 revealed that strains TN-271T and TN-272 were distinct from C. scindens JCM 6567T. On the basis of phylogenetic analysis and DNA-DNA similarity data, it was concluded that strains TN-271T and TN-272 are members of a new species of the genus Clostridium, for which the name Clostridium hylemonae is proposed. The type strain is strain TN-271T (= JCM 10539T). PMID- 10843035 TI - Bulleidia extructa gen. nov., sp. nov., isolated from the oral cavity. AB - Five strains of anaerobic non-sporing Gram-positive bacilli isolated from advanced periodontitis (four strains) and a dentoalveolar abscess (one strain) that did not correspond to existing species were subjected to phenotypic and genetic characterization. Following 16S rDNA sequence analysis, they were found to constitute a novel branch of the low G+C Gram-positive division of the phylogenetic tree related to Erysipelothrix rhusiopathiae and Holdemania filiformis. A new genus Bulleidia, and the species Bulleidia extructa, are proposed. Growth of B. extructa in broth media was poor but was enhanced by the addition of fructose, glucose or maltose together with Tween 80. Glucose and maltose were fermented and arginine was hydrolysed. Acetate, lactate and trace amounts of succinate were the end products of glucose fermentation. The G+C content of the DNA of the type strain is 38 mol%. The type strain of Bulleidia extructa is DSM 13220T. PMID- 10843036 TI - Rhodopseudomonas rhenobacensis sp. nov., a new nitrate-reducing purple non-sulfur bacterium. AB - During the course of isolating and identifying purple non-sulfur bacteria, one nitrate-reducing strain was isolated which did not fit the description of any other purple non-sulfur bacterium known to date. The isolate had rod-shaped cells that contained lamellar intracytoplasmic membranes and produced red cultures. Absorption maxima of photosynthetically grown cell homogenates were at 376, 471, 503, 540, 591, 805 and 878 nm. The new isolate grew anaerobically in the light or aerobically in the dark. Various organic compounds served as carbon sources and electron donors. The predominant quinone was ubiquinone 10, the predominant fatty acid was 18:1omega7c. The polar lipids comprised diphosphatidyl glycerol, phosphatidyl glycerol, phosphatidyl ethanolamine and phosphatidyl choline. Analysis of the 16S rDNA gene sequences revealed that the new isolate was closely related to Rhodopseudomonas palustris. A DNA-DNA- hybridization study differentiated the new isolate and Rhodopseudomonas palustris at the species level. Therefore, the name Rhodopseudomonas rhenobacensis sp. nov. is proposed for the new isolate. PMID- 10843037 TI - Cellulomonas persica sp. nov. and Cellulomonas iranensis sp. nov., mesophilic cellulose-degrading bacteria isolated from forest soils. AB - Two newly described species of mesophilic, cellulose-degrading, aerobic bacteria were isolated from forest humus soils along the southern border of the Caspian Sea. Cellulomonas persica and Cellulomonas iranensis are proposed as new specific epithets based on comparative sequence analyses of 16S rDNA, DNA-DNA hybridization and phenotypic characteristics. Formal species descriptions are provided. PMID- 10843038 TI - Ferroplasma acidiphilum gen. nov., sp. nov., an acidophilic, autotrophic, ferrous iron-oxidizing, cell-wall-lacking, mesophilic member of the Ferroplasmaceae fam. nov., comprising a distinct lineage of the Archaea. AB - An isolate of an acidophilic archaeon, strain YT, was obtained from a bioleaching pilot plant. The organism oxidizes ferrous iron as the sole energy source and fixes inorganic carbon as the sole carbon source. The optimal pH for growth is 1.7, although growth is observed in the range pH 1.3 to 2.2. The cells are pleomorphic and without a cell wall. 16S rRNA gene sequence analysis showed this strain to cluster phylogenetically within the order 'Thermoplasmales' sensu Woese, although with only 89.9 and 87.2% sequence identity, respectively, to its closest relatives, Picrophilus oshimae and Thermoplasma acidophilum. Other principal differences from described species of the 'Thermoplasmales' are autotrophy (strain YT is obligately autotrophic), the absence of lipid components typical of the ' Thermoplasmales' (no detectable tetraethers) and a lower temperature range for growth (growth of strain YT occurs between 15 and 45 degrees C). None of the sugars, amino acids, organic acids or other organic compounds tested was utilized as a carbon source. On the basis of the information described above, the name Ferroplasma acidiphilum gen. nov., sp. nov. is proposed for strain YT within a new family, the Ferroplasmaceae fam. nov. Strain YT is the type and only strain of F. acidiphilum. This is the first report of an autotrophic, ferrous-iron-oxidizing, cell-wall-lacking archaeon. PMID- 10843039 TI - Analysis of the actinobacterial insertion in domain III of the 23S rRNA gene of uncultured variants of the bacterium associated with Whipple's disease using broad-range and 'Tropheryma whippelii'-specific PCR. AB - Heterogeneity in the 16S-23S rDNA spacer of uncultured 'Tropheryma whippelii' has previously been reported. In this study, the hypervariable insertion in the 23S rDNA domain III characteristic for actinobacteria was analysed. The finding of a unique sequence virtually identical among the subtypes supports the classification of 'T. whippelii' among the actinobacteria and the concept of three subtypes rather than three subspecies, and provides an alternative target for diagnostic assays. PMID- 10843040 TI - Application of AFLP for taxonomic and epidemiological studies of Photobacterium damselae subsp. piscicida. AB - A collection of 106 Photobacterium damselae subsp. piscicida strains and 19 Photobacterium damselae subsp. damselae strains, including reference and type strains, were genetically characterized using AFLP. The total genomic DNA of each bacterial strain was digested using restriction endonucleases HindIII and TaqI. Using numerical analysis, six clusters were recognized. The largest cluster (n = 106) contained the majority of the strains tested and consisted exclusively of Photobacterium damselae subsp. piscicida. The Photobacterium damselae subsp. damselae strains fell outside this cluster. DNA-DNA hybridization experiments showed 77% DNA binding between the two subspecies, indicating a close genetic relationship. This clearly demonstrates the applicability of AFLP in studying the taxonomic position of Photobacterium damselae subsp. piscicida. In addition, AFLP proved to be a useful genotypic technique for epidemiological surveys of the pathogen, since it was able to discriminate between Mediterranean and Japanese Photobacterium damselae subsp. piscicida isolates. PMID- 10843041 TI - Protein-coding genes as molecular markers for ecologically distinct populations: the case of two Bacillus species. AB - Bacillus globisporus and Bacillus psychrophilus are one among many pairs of ecologically distinct taxa that are distinguished by very few nucleotide differences in 16S rRNA gene sequence. This study has investigated whether the lack of divergence in 16S rRNA between such species stems from the unusually slow rate of evolution of this molecule, or whether other factors might be preventing neutral sequence divergence at 16S rRNA as well as every other gene. B. globisporus and B. psychrophilus were each surveyed for restriction-site variation in two protein-coding genes. These species were easily distinguished as separate DNA sequence clusters for each gene. The limited ability of 16S rRNA to distinguish these species is therefore a consequence of the extremely slow rate of 16S rRNA evolution. The present results, and previous results involving two Mycobacterium species, demonstrate that there exist closely related species which have diverged long enough to have formed clearly separate sequence clusters for protein-coding genes, but not for 16S rRNA. These results support an earlier argument that sequence clustering in protein-coding genes could be a primary criterion for discovering and identifying ecologically distinct groups, and classifying them as separate species. PMID- 10843042 TI - Cellular fatty acid composition of cyanobacteria assigned to subsection II, order Pleurocapsales. AB - The cellular fatty acid composition of five of the six genera of unicellular cyanobacteria in subsection II, Pleurocapsales (Dermocarpa, Xenococcus, Dermocarpella, Myxosarcina and the Pleurocapsa assemblage) contained high proportions of saturated straight-chain fatty acids (26-41% of the total) and unsaturated straight chains (40-67%). Isomers of 16:1 were the main monounsaturated acid component (11-59%). Polyunsaturated acids were present at trace levels (0-1% or less) in Xenococcus and Myxosarcina, at concentrations of less than 7% in Dermocarpa, Dermocarpella, Pleurocapsa and CCMP 1489, and at high concentrations (35% or more) in Chroococcidiopsis. Chroococcidiopsis was also different in terms of the percentage of 16:1 isomers (10-12%) compared to other genera (30-59%), and in terms of total 16-carbon and 18-carbon fatty acids. In general, the composition and heterogeneity of fatty acids in the order Pleurocapsales was similar to that reported for the unicellular cyanobacteria of subsection I, order Chroococcales. PMID- 10843043 TI - Reductively debrominating strains of Propionigenium maris from burrows of bromophenol-producing marine infauna. AB - Two novel strains of Propionigenium maris able to reductively debrominate 2,4,6 tribromophenol (TBP) to monobromophenols were isolated from marine hemichordate and polychaete burrows. These two strains, DSL-1 and ML-1, were anaerobic, non motile rods that stained Gram-negative and required 0.05% yeast extract for growth. Strain DSL-1 fermented pyruvate and succinate to predominantly butyrate and strain ML-1 fermented glucose and succinate primarily to propionate. No inorganic terminal electron acceptors were identified. The pH and temperature optima for growth were 7.6 and 30 degrees C for strain DSL-1 and 7.0 and 32 degrees C for strain ML-1, respectively; doubling times for strains DSL-1 and ML 1 were 0.32 h and 0.30 h, respectively. Both strains required 2-3% (w/v) NaCl for optimal growth. Morphological and physiological features, as well as the results of 16S rDNA sequence analysis, showed these to be new strains of Propionigenium maris. Because they differ from the P. maris type strain (DSM 9537T) in a number of respects, including their ability to rapidly debrominate di- and tribromophenols, and in their specific habitats, the species description is amended to include these ecologically important properties. PMID- 10843044 TI - Characterization of Nodularia strains, cyanobacteria from brackish waters, by genotypic and phenotypic methods. AB - An investigation was undertaken of the genetic diversity of Nodularia strains from the Baltic Sea and from Australian waters, together with the proposed type strain of Nodularia spumigena. The Nodularia strains were characterized by using a polyphasic approach, including RFLP of PCR-amplified 16S rRNA genes, 16S rRNA gene sequencing, Southern blotting of total DNA, repetitive extragenic palindromic- and enterobacterial repetitive intergenic consensus-PCR, ribotyping and phenotypic tests. With genotypic methods, the Nodularia strains were grouped into two clusters. The genetic groupings were supported by one phenotypic property: the ability to produce nodularin. In contrast, the cell sizes of the strains were not different in the two genetic clusters. 16S rRNA gene sequences indicated that all the Nodularia strains were closely related, despite their different origins. According to this study, two genotypes of Nodularia exist in the Baltic Sea. On the basis of the taxonomic definitions of Komarek et al. (Algol Stud 68, 1-25, 1993), the non-toxic type without gas vesicles fits the description of Nodularia sphaerocarpa, whereas the toxic type with gas vesicles resembles the species N. spumigena and Nodularia baltica. PMID- 10843045 TI - Taxonomy of Antarctic Flavobacterium species: description of Flavobacterium gillisiae sp. nov., Flavobacterium tegetincola sp. nov., and Flavobacterium xanthum sp. nov., nom. rev. and reclassification of [Flavobacterium] salegens as Salegentibacter salegens gen. nov., comb. nov. AB - 16S rRNA phylogenetic analysis of a number of yellow- and orange-pigmented strains isolated from a variety of Antarctic habitats including sea ice, lakewater and cyanobacterial mats indicated a close relationship to the genus Flavobacterium but distinct from known Flavobacterium species. Phenotypic properties, DNA G+C content and whole-cell fatty acid profiles of the Antarctic strains were consistent with those of the genus Flavobacterium. DNA-DNA hybridization analysis indicated the presence of two distinct and novel genospecies each isolated from a different Antarctic habitat. From polyphasic taxonomic data it is proposed that the two groups represent new species with the following proposed names: Flavobacterium gillisiae (ACAM 601T) and Flavobacterium tegetincola (ACAM 602T). In addition polyphasic analysis of the species '[Cytophaga] xantha' (Inoue and Komagata 1976), isolated from Antarctic mud, indicated it was a distinct member of the genus Flavobacterium and was thus revived as Flavobacterium xanthum. Phylogenetic and fatty acid analyses also indicate that the species [Flavobacterium] salegens (Dobson et al. 1993), from Organic Lake, Antarctica, is misclassified at the genus level. It is proposed that this species belongs to a new genus, Salegentibacter salegens gen. nov., comb. nov. PMID- 10843046 TI - Haloterrigena thermotolerans sp. nov., a halophilic archaeon from Puerto Rico. AB - An extremely halophilic Archaeon belonging to the order Halobacteriales was isolated from the solar salterns of Cabo Rojo, Puerto Rico. The organism, designated strain PR5T, is rod-shaped, non-motile and requires at least 12% (w/v) NaCl to grow. The strain is highly thermotolerant: its temperature optimum is 50 degrees C and growth is possible up to 60 degrees C. Polar lipid analysis revealed the presence of the bis-sulfated glycolipid S2-DGD-1 as sole glycolipid and the absence of the glycerol diether analogue of phosphatidylglycerosulfate. Both C20,C20 and C20,C25 core lipids are present. The G+C content of the DNA is 63.3 mol%. According to 16S rDNA sequence data, strain PR5T is closely related to the representatives of the genera Haloterrigena and Natrinema, but on the basis of its phenotypic properties, 16S rDNA sequence and DNA-DNA hybridization studies, strain PR5T cannot be assigned to any of the recognized species within these genera. On the basis of its polar lipid composition, the isolate has been assigned to the genus Haloterrigena. The creation of a new species, Haloterrigena thermotolerans, is therefore proposed to accommodate this isolate. The type strain is strain PR5T (= DSM 11552T = ATCC 700275T). PMID- 10843047 TI - The Streptococcus anginosus species comprises five 16S rRNA ribogroups with different phenotypic characteristics and clinical relevance. AB - A collection of 267 consecutively isolated Streptococcus anginosus strains was screened for the prevalence of previously described 'motile' strains by hybridization with oligonucleotide probes in a reverse line blot assay. The motile strains represented 101 (37.8%) of the S. anginosus strains. The vast majority of these strains fermented mannitol and raffinose, whereas most other S. anginosus strains did not (P<0.001). Most (83/101) of the motile strains were recovered from the urogenital tract (including five strains from neonatal surveillance cultures) and only a minority (36) of them were associated with infection-related samples (P<0.001). Strains that hybridized with the same oligonucleotide probes as the type strain S. anginosus ATCC 33397T (= NCTC 10713T) were designated ATCC-like strains. They accounted for 74 (27.7%) of the strains examined, were commonly distributed among the different body parts and were significantly more associated with infection-related samples. Three other hybridization patterns were recognized in the reverse line blot assay, ribogroup I (n = 51), ribogroup II (n = 21) and ribogroup III (n = 19). Ribogroup II strains were significantly more frequently recovered from the abdominal cavity and were associated with infection-related samples. Ribogroup I included the majority of the S. anginosus strains that carried Lancefield group C. Comparison of the nearly complete 16S rRNA sequence of two representative strains of each ribogroup revealed that all five ribogroups were closely related (>97% sequence similarities), and that most sequence divergences between the ribogroups occurred in the 1024-1064 bp region of the 16S rRNA gene. The present data confirm the heterogeneity within the S. anginosus species. PMID- 10843048 TI - Phylogenetic analysis of the genus Thermoactinomyces based on 16S rDNA sequences. AB - The almost complete 16S rDNA sequences of the type strains of eight validly described species and two invalid species of the genus Thermoactinomyces were determined and phylogenetically analysed. The validly described Thermoactinomyces species formed phylogenetic lineages related to the family Bacillaceae, as shown previously. However, the available strains of 'Thermoactinomyces glaucus' and 'Thermoactinomyces monosporus' exhibited their closest phylogenetic affinities not to the genus Thermoactinomyces but to the genus Saccharomonospora. Some Thermoactinomyces species were shown to be closely related from 16S rDNA sequence analysis. Particularly, Thermoactinomyces vulgaris KCTC 9076T and Thermoactinomyces candidus KCTC 9557T had the same 16S rDNA sequences and Thermoactinomyces thalpophilus KCTC 9789T and Thermoactinomyces sacchari KCTC 9790T showed 16S rDNA similarity value of almost 100%. From phylogenetic analysis based on 16S rDNA sequences, it is suggested that the genus Thermoactinomyces should be taxonomically re-evaluated using other useful taxonomic markers. PMID- 10843049 TI - Rapid identification of Staphylococcus epidermidis. AB - During the collection of airborne bacteria in a museum in England some bacterial strains were isolated which due to their fatty acid profiles were clearly identified as members of the genus Staphylococcus. As fatty acid compositions of coagulase-negative staphylococci are very similar, differing only in quantities but not in qualities, further identification at the species level without a fatty acid database was not achieved. Investigation of the isolates using the Staph ID 32 API system resulted in an identification of the isolates as Staphylococcus epidermidis (probabilities of 79.7-95.5%). For further genotypic characterization of these isolates, some Staphylococcus epidermidis strains from different sources and the type strains of Staphylococcus aureus, Staphylococcus capitis, Staphylococcus epidermidis, Staphylococcus gallinarum, Staphylococcus haemolyticus, Staphylococcus hominis, Staphylococcus warneri and Staphylococcus xylosus were subjected to repetitive-sequence PCR, including enterobacterial repetitive intergenic consensus (ERIC) PCR, BOX-PCR and repetitive extragenic palindromic unit sequence (REP) PCR. ERIC- and BOX-PCR yielded a species-specific banding pattern for all Staphylococcus epidermidis strains. Furthermore, all staphylococcal reference strains investigated exhibited distinct banding patterns, clearly distinguishable from that of Staphylococcus epidermidis. No species-specific banding patterns could be observed after REP-PCR. As species identification of coagulase-negative staphylococci by fatty acid analyses and biochemical tests is known to be difficult ERIC- and BOX-PCR seem to be excellent tools for the identification of Staphylococcus epidermidis isolates. PMID- 10843050 TI - DNA-DNA hybridization determined in micro-wells using covalent attachment of DNA. AB - The present study was aimed at reducing the time and labour used to perform DNA DNA hybridizations for classification of bacteria at the species level. A micro well-format DNA hybridization method was developed and validated. DNA extractions were performed by a small-scale method and DNA was sheared mechanically into fragments of between 400 and 700 bases. The hybridization conditions were calibrated according to DNA similarities obtained by the spectrophotometric method using strains within the family Pasteurellaceae. Optimal conditions were obtained with 300 ng DNA added per well and bound by covalent attachment to NucleoLink. Hybridization was performed with 500 ng DNA, 5% (w/w) of which was labelled with photo-activatable biotin (competitive hybridization) for 2.5 h at 65 degrees C in 2 x SSC followed by stringent washing with 2 x SSC at the same temperature. The criteria for acceptance of results were a maximum of 15% standard deviation, calculated as a percentage of the mean for four replicate micro-wells, and that DNA similarities were not significantly different in at least two independent experiments. The relationship between DNA similarities obtained by the micro-well method (y) and by the spectrophotometric method (x) was y = 0.534x+30.6, when these criteria had been applied to 23 pairs of strains of Actinobacillus species, avian [Pasteurella] haemolytica-like bacteria and Mannheimia species. The correlation (Pearson) between DNA similarities obtained by interchange of strains used for covalent binding and hybridization was 0.794. Significantly lower DNA similarities were observed by the spectrophotometric compared with the micro-well method for three pairs of hybridizations. After removal of these data, the relationship between DNA similarities obtained by the micro-well and spectrophotometric methods improved to y = 0.855x + 11.0. It was found that the accuracy and precision of the micro-well method was at the same level as that of the spectrophotometric method, but the labour and analysis time were reduced significantly. The use of hybridization in the micro-well format will allow DNA-DNA hybridizations to be carried out between all strains selected for a particular taxonomic study, in order to construct complete data matrices and improve species definition. PMID- 10843051 TI - Catellatospora koreensis sp. nov., a novel actinomycete isolated from a gold-mine cave. AB - A new actinomycete strain, LM 042T, which was isolated from a gold-mine cave in Kongju, Republic of Korea, is described by phenotypic and genotypic characters. The organism formed short chains of non-motile spores and globose bodies from substrate mycelium. An aerial mycelium was absent. This organism was chemotaxonomically characterized by the presence of meso-diaminopimelic acid, rhamnose, xylose, glucose, mannose and ribose in whole-cell hydrolysates (a type II cell wall and a variant of sugar pattern D), a glycolyl type of muramic acid, DNA G+C content of 70.4 mol%, a type PII phospholipid pattern (phosphatidylethanolamine as a diagnostic nitrogenous phospholipid), a tetrahydrogenated menaquinone with 10 isoprene units as a major menaquinone, and fatty acid profiles predominated by iso-branched hexadecanoic acid, iso-branched pentadecanoic acid and heptadcenoic acid. A comparative analysis of 16S rDNA sequences indicated that this organism formed a distinct clade within the evolutionary radiation of the family Micromonosporaceae and clustered with members of the genus Catellatospora. The 16S rDNA similarity values between the isolate and its phylogenetic neighbours, the two subspecies of Catellatospora citrea and Catellatospora tsunoense, were 95.0-95.2% and 94.9%, respectively. An equidistant relationship was observed among the isolate, Catellatospora ferruginea and all other members of the Micromonosporaceae genera (levels of similarity 93.0-94.0%). The combination of physiological, chemotaxonomic and DNA DNA hybridization data supported that this organism is a novel species of the genus Catellatospora, for which the name Catellatospora koreensis sp. nov. is proposed. The type strain is LM 042T (= IMSNU 50729T). PMID- 10843052 TI - Emendation of the description of Blastomonas natatoria (Sly 1985) Sly and Cahill 1997 as an aerobic photosynthetic bacterium and reclassification of Erythromonas ursincola Yurkov et al. 1997 as Blastomonas ursincola comb. nov. AB - Photosynthetic properties of Blastomonas natatoria (Sly 1985) Sly and Cahill 1997, which had been recognized as being non-photosynthetic, were examined and compared with those of its close relative, the aerobic photosynthetic bacterium, Erythromonas ursincola Yurkov et al. 1997. HPLC experiments demonstrated that bacteriochlorophyll a was present in a detectable amount in the lipid extract from B. natatoria DSM 3183T as well as that from E. ursincola DSM 9006T. The puf genes, encoding the proteins of the photosynthetic reaction centre and core light harvesting complexes, were detected by PCR from both the organisms. 16S rDNA sequence comparisons and DNA-DNA hybridization studies confirmed that B. natatoria and E. ursincola were closely related genetically in a single genus. On the basis of phenotypic, chemotaxonomic and phylogenetic data, it is proposed that the description of B. natatoria is emended as a species of aerobic photosynthetic bacteria and that E. ursincola is reclassified as Blastomonas ursincola comb. nov. PMID- 10843053 TI - Aeromonas salmonicida subsp. pectinolytica subsp. nov., a new pectinase-positive subspecies isolated from a heavily polluted river. AB - Aeromonas strains which phenotypically and genetically belong to the Aeromonas salmonicida species but that according to their phenotypic properties constitute a new subspecies have been isolated from the water of a heavily polluted river, the Matanza river, situated near the central district of Buenos Aires city. These strains were ascribed to the A. salmonicida species by using 65 biochemical tests and by DNA-DNA hybridization. They produce acid from -sorbitol, an unusual biochemical property found in a few members of the A. salmonicida species. They also utilize urocanic acid and do not ferment L-rhamnose or utilize LD-lactate, and are elastase- and gluconate-negative. The DNA relatedness was over 70%, the current limit accepted for the phylogenetic definition of a species, to the described A. salmonicida subspecies and nearly 100% within the new group of Aeromonas strains. Phenotypic differentiation from other A. salmonicida subspecies was readily achieved using the following characteristics: growth at 37 degrees C, melanin production, indole and Voges-Proskauer assays, growth on KCN broth, mannitol and sucrose fermentation and gas from glucose. A remarkable property of the strains of the new group was their ability to degrade polypectate, an unusual feature among Aeromonas species in general. The complete 16S rRNA gene of one strain of the new group was sequenced. Comparison with rDNA sequences of Aeromonas members available in databases revealed a close relationship between this strain and strains belonging to A. salmonicida subsp. salmonicida, masoucida and achromogenes, in agreement with the biochemical data. Since the new A. salmonicida strains constitute a tight genomic group that can be identified by phenotypic properties it was concluded that they represent a new subspecies for which the name Aeromonas salmonicida subsp. pectinolytica is proposed. The type strain of A. salmonicida subsp. pectinolytica is 34melT (= DSM 12609T). PMID- 10843054 TI - Acholeplasma vituli sp. nov., from bovine serum and cell cultures. AB - Organisms isolated from commercial foetal bovine serum and from cell culture lines containing such serum supplements were found to consist of non-helical, non motile, pleomorphic coccoid forms. One strain (FC 097-2T) cultivated directly from foetal bovine serum was selected for characterization. In ultrastructural examination, individual round cells lacked cell wall structures and cells varied in size, with a mean diameter of about 700 nm. However, variable numbers of cells were filterable through membranes of 300 nm. Optimum growth occurred between 30 and 37 degrees C. The organism fermented glucose, fructose and mannose, but did not hydrolyse arginine. The strain was insensitive to 500 U penicillin ml(-1) and was capable of growing in the absence of serum or cholesterol. The organism was serologically distinct from all 13 currently described species in the genus Acholeplasma and from other sterol-requiring species in the genus Mycoplasma, using growth inhibition, immunoperoxidase and immunofluorescence tests. Strain FC 097-2T was found to have a DNA G+C composition between 37.6 +/- 1 mol% and 38.3 +/- 1 mol%. The genome size was determined to be 2095 kbp. The 16S rDNA sequence of strain FC 097-2T was compared to 16S rDNA sequences of other mollicutes in nucleotide databases. No deposited sequence was found to be identical; the closest relatives were several members of the genus Acholeplasma. On the basis of these findings and other similarities to acholeplasmas in morphology and growth, the absence of a sterol requirement for growth, and similar genomic characteristics, the organism was assigned to the genus Acholeplasma. Strain FC 097-2T is designated the type strain (ATCC 700667T) of a new species, Acholeplasma vituli. PMID- 10843055 TI - Polyphasic taxonomic approach in the description of Alishewanella fetalis gen. nov., sp. nov., isolated from a human foetus. AB - A taxonomically unique bacterium is described on the basis of a physiological and biochemical characterization, fatty acid profiling and sequence analyses of 16S rRNA and gyrase B (gyrB) genes. This non-motile, non-fermentative bacterium was isolated from a human foetus in Uppsala, Sweden, and originally misidentified as a Shewanella putrefaciens by conventional biochemical testing. The bacterium grew well at mesophilic temperatures with optimum growth at 37 degrees C. It was facultatively anaerobic and utilized various electron acceptors (trimethylamine oxide, nitrate, nitrite and thiosulphate). The dominant fatty acids were 17:1B, 16:1 cis9, 17:0 and 16:0. Fatty acids 13:0 iso and 15:0 iso, which have been found to be typical of Shewanella species were not detected. The G+C content of the DNA was 50.6 mol%. Phylogenetic analysis of the 16S rRNA gene sequence revealed a clear affiliation with members of the gamma subclass of the Proteobacteria. No relationship was seen with any of the established genera in the gamma subclass of the Proteobacteria, although a distinct relationship with Vibrionaceae was observed. That the bacterium represents a novel bacterial genus distinct from Vibrionaceae was also supported by gyrB sequence analysis. Considering the source and close proximity to the genus Shewanella, the name Alishewanella fetalis gen. nov., sp. nov. is proposed, for which the type strain is strain CCUG 30811T. PMID- 10843056 TI - Lactobacillus algidus sp. nov., a psychrophilic lactic acid bacterium isolated from vacuum-packaged refrigerated beef. AB - Lactobacillus algidus sp. nov. is described on the basis of 40 strains isolated as one of the predominant bacteria from five specimens of vacuum-packaged beef collected from different meat shops and stored at 2 degrees C for 3 weeks. These strains were quite uniform in the overall characteristics examined. They are facultatively anaerobic, psychrophilic, Gram-positive, non-spore-forming, non motile, lactic acid-homofermentative rods. The cells occurred singly and in pairs on agar media and in rather long chains in broth media. They differed in several cultural and biochemical characteristics from the authentic meso-diaminopimelic acid-positive or psychrophilic lactic acid bacteria in the genera Lactobacillus, Carnobacterium and Brochothrix. The SDS-PAGE whole-cell protein pattern was clearly distinctive. DNA-DNA hybridization and phylogenetic analysis of 16S rDNA also failed to associate these strains closely with any of the validly described organisms used. The phylogenetic analysis showed that these strains are rather remotely but most closely related to Lactobacillus mali (93% sequence similarity), which belongs to the Lactobacillus casei/Pediococcus group. Therefore, these strains should be included in the genus Lactobacillus and considered to represent a new species, Lactobacillus algidus sp. nov. The type strain is M6A9T (= JCM 10491T). PMID- 10843057 TI - Vagococcus fessus sp. nov., isolated from a seal and a harbour porpoise. AB - A polyphasic taxonomic study was performed on two strains of an unknown Gram positive, catalase-negative, coccus-shaped bacterium isolated from a dead seal and a harbour porpoise. Comparative 16S rRNA gene sequencing demonstrated that the unknown bacterium represents a new subline within the genus Vagococcus close to, but distinct from, Vagococcus fluvialis, Vagococcus lutrae and Vagococcus salmoninarum. The unknown bacterium was readily distinguished from the three currently recognized Vagococcus species by biochemical tests and electrophoretic analysis of whole-cell proteins. Based on phylogenetic and phenotypic evidence, it is proposed that the unknown bacterium be classified as a new species, Vagococcus fessus. The type strain of Vagococcus fessus is CCUG 41755T. PMID- 10843058 TI - Caloramator coolhaasii sp. nov., a glutamate-degrading, moderately thermophilic anaerobe. AB - An obligately anaerobic, moderately thermophilic, glutamate-degrading bacterium (strain ZT) was isolated from an enrichment culture obtained from anaerobic thermophilic granular sludge. The cells were rod-shaped to filamentous and showed no motility or spore formation. The cell wall had a Gram-positive structure, which was revealed by electron microscopy. Optimum growth of the strain was observed under neutrophilic conditions at 50-55 degrees C. The doubling time of strain ZT grown in rich medium was approximately 1 h at optimal pH and temperature. Strain ZT was able to grow on a variety of organic compounds. Most carbon sources were converted to acetate, CO2, H2, and traces of propionate and lactate. Strain ZT oxidized glutamate to acetate, CO2, NH4+, traces of propionate and H2. The doubling time on this substrate was 1-6 d. The strain fermented glutamate syntrophically in co-culture with Methanobacterium thermoautotrophicum Z-245T to the same products, but the co-culture had a fourfold higher growth rate. 16S rDNA sequence analysis revealed a relationship with Thermobrachium celere, Caloramator indicus and Caloramator proteoclasticus. The G+C content was 31.7 mol%. Based on its morphological, phylogenetic and physiological characteristics, it is proposed that strain ZT should be classified in the genus Caloramator as a new species, Caloramator coolhaasii. PMID- 10843059 TI - Acrocarpospora gen. nov., a new genus of the order Actinomycetales. AB - The taxonomic position of two actinomycete strains isolated from soil was studied. The isolates contained glutamic acid, alanine and meso-diaminopimelic acid as cell-wall amino acids and menaquinone MK-9(H4) and madurose in the whole cell hydrolysate. Phylogenetic analysis revealed that the isolates belonged to the family Streptosporangiaceae, but not to any known genus, and formed a monophyletic cluster with Streptosporangium corrugatum. On the basis of morphological characteristics, phylogenetic analysis and DNA-DNA hybridization, the name Acrocarpospora gen. nov. is proposed for a new genus containing the isolates and Streptosporangium corrugatum, and Acrocarpospora pleiomorpha sp. nov. R-31T (= IFO 16267T), Acrocarpospora macrocephala sp. nov. R-55T (=IFO 16266T) and Acrocarpospora corrugata comb. nov. IFO 13972T are described. PMID- 10843060 TI - An unusual Streptococcus from human urine, Streptococcus urinalis sp. nov. AB - Biochemical, molecular chemical and molecular genetic studies were performed on an unknown Gram-positive, catalase-negative, chain-forming coccus isolated from the urine of a patient suffering from cystitis. Comparative 16S rRNA gene sequencing showed that the organism is a member of the 'pyogenic subgroup' of the genus Streptococcus and has a close affinity with Streptococcus pyogenes and Streptococcus canis. The unknown coccus was, however, readily distinguished from these species and other streptococci by biochemical tests and electrophoretic analysis of whole-cell proteins. Based on phenotypic and phylogenetic evidence, it is proposed that the unknown bacterium be classified as a new species of the genus Streptococcus, Streptococcus urinalis sp. nov. The type strain of Streptococcus urinalis is CCUG 41590T. PMID- 10843061 TI - Relationships of bradyrhizobia from Platypodium and Machaerium (Papilionoideae: tribe Dalbergieae) on Barro Colorado Island, Panama. AB - Enzyme electrophoresis and rRNA sequencing indicated that root nodule bacteria from the canopy tree Platypodium elegans and the lianas Machaerium milleflorum and Machaerium arboreum on Barro Colorado Island, Panama, were highly diverse on a local scale. A total of 11 distinct multilocus genotypes [ETs (electrophoretic types)] was found among the 33 isolates analysed. On average, ETs differed from one another at 74% of the 11 enzyme loci assayed, and separate nodules on a single host individual were often occupied by genetically divergent ETs. Certain ETs were sampled multiple times from both Platypodium and Machaerium, suggesting a lack of specificity toward the two legume genera. Within the intervening sequence (IVS) region in the 5' end of 23S rRNA, seven ETs had a length variant similar to that of Bradyrhizobium japonicum USDA 110, and the other four ETs had an IVS region 26-28 bp shorter. Parsimony analysis of both partial 23S rRNA and nearly full-length 16S rRNA sequences indicated that all Platypodium and Machaerium isolates were related to B. japonicum rather than Bradyrhizobium elkanii. The 16S rRNA sequence of one isolate was >99% similar to that of B. japonicum USDA 110, and the closest known relatives for other isolates were Philippine bradyrhizobia from the legumes Stylosanthes and Samanea. PMID- 10843062 TI - Sequence analysis of domains III and IV of the 23S rRNA gene of verticillate streptomycetes. AB - Domains III and IV of the 23S rRNA gene of 25 strains of verticillate streptomycetes were sequenced. None of the sequences was identical to any other, with regions of variability being restricted to parts of helices 54 and 64. No relationships were detected between the similarities in the sequence and the assignment to phenetic clusters as defined by the numerical taxonomy studies. Limited agreement was also found between similarity of the sequences and DNA-DNA homology values. However, species (> 70% DNA-DNA homology values)-specific diagnostic oligonucleotides generally could be defined, except for Streptomyces baldaccii. Therefore the determination of the 23S rRNA sequence may be of greater value for fingerprinting individual strains than for taxonomic or identification purposes. PMID- 10843063 TI - Rhodococcus koreensis sp. nov., a 2,4-dinitrophenol-degrading bacterium. AB - A 2,4-dinitrophenol-degrading bacterial strain, DNP505T, which was isolated from an industrial wastewater, was taxonomically studied by a polyphasic approach using phenotypic, chemotaxonomic and genetic methods. Strain DNP505T has a cell wall of chemotype IV containing meso-diaminopimelic acid, arabinose and galactose. The predominant menaquinone is MK-8(H2). Mycolic acids contain 43-53 carbon atoms. Strain DNP505T has a cellular fatty acid profile containing straight-chain saturated, unsaturated and 10-methyl-branched fatty acids and has C16:0 as the major fatty acid. The DNA G+C content is 66 mol%. Strain DNP505T formed a coherent cluster with Rhodococcus species in a phylogenetic inference based on 16S rDNA sequences. Interestingly, strain DNP505T was found to have two types of 16S rDNA sequence, which showed 10 bp sequence differences (99.3% nucleotide similarity). Its differences in some phenotypic characteristics and its genetic distinctiveness indicate that strain DNP505T is separate from Rhodococcus species described previously. It is therefore proposed that strain DNP505T should be placed in the genus Rhodococcus as a new species, Rhodococcus koreensis. The type strain of the new species is strain DNP505T (= KCTC 0569BPT = JCM 10743T). PMID- 10843064 TI - Gordonia nitida sp. nov., a bacterium that degrades 3-ethylpyridine and 3 methylpyridine. AB - A bacterial strain, LE31T, which is capable of degrading 3-ethylpyridine and 3 methylpyridine, was isolated from an industrial wastewater and was taxonomically studied by using a polyphasic approach. Strain LE31T was identified as a member of the genus Gordonia on the basis of chemotaxonomic characteristics and phylogenetic inference-based 16S rDNA sequence. The cell wall contained meso diaminopimelic acid, arabinose and galactose (wall chemotype IV). The predominant menaquinone was MK-9(H2). The mycolic acids contained 47-55 carbon atoms. The major fatty acids were C16:0, C18:1 omega9c, 10-methyl-C18:0 (TBSA). The G+C content of DNA was 67 mol%. The 16S rDNA sequence of strain LE31T was most similar to that of the type strain of Gordonia rubropertincta. The differences in some phenotypic characteristics and the genetic distinctiveness distinguish strain LE31T from the Gordonia species described previously. Therefore it is proposed that strain LE31T should be placed in the genus Gordonia as a new species. The name Gordonia nitida is proposed for strain LE31T. The type strain of the new species is strain LE31T (= KCTC 0605BPT = KCCM 80004T). PMID- 10843065 TI - Systematic analysis of xanthomonads (Xanthomonas spp.) associated with pepper and tomato lesions. AB - The taxonomy and evolutionary relationships among members of the genus Xanthomonas associated with tomato and pepper have been a matter of considerable controversy since their original description in 1921. These bacteria, which are a major affliction of tomato and pepper crops in warm and humid regions, were originally described as a single species, but subsequent research has shown the existence of at least two genetic groups differentiated by physiological, biochemical and pathological characteristics. This work synthesizes the findings from several approaches, including pathogenicity tests, enzymic activity, restriction fragment analysis of the entire genome, DNA-DNA hybridization and RNA sequence comparisons based on a 2097 base sequence comprising the 16S rRNA gene, the intergenic spacer located between the 16S and 23S rRNA genes and a small region of the 23S rRNA gene. Within the group of xanthomonads pathogenic on pepper and tomato four distinct phenotypic groups exist, of which three form distinct genomic species. These include Xanthomonas axonopodis pv. vesicatoria (A and C group), Xanthomonas vesicatoria (B group) and Xanthomonas gardneri (D group). On the basis of phenotypic and genotypic differences between A- and C group strains, the C strains should be considered as a subspecies within Xanthomonas axonopodis pv. vesicatoria. PMID- 10843066 TI - Papillibacter cinnamivorans gen. nov., sp. nov., a cinnamate-transforming bacterium from a shea cake digester. AB - A new, strictly anaerobic, Gram-positive, non-sporulating, mesophilic bacterium, designated strain CIN1T (T=type strain) was isolated from an anaerobic digester fed with shea cake rich in tannins and aromatic compounds. Cells of strain CIN1T were rod-shaped, had characteristically pointed ends (1.3-3.0 x 0.5-0.6 microm) and occurred singly, in pairs and sometimes in chains of up to six. The pH range for growth was 6.9-8.5 and the temperature growth range was 15-40 degrees C. Optimum growth occurred with yeast extract and cinnamate at 37 degrees C and a pH of 7.5. The isolate transformed cinnamate by degrading the aliphatic side chain to produce acetate and benzoate rather than by aromatic ring cleavage or demethoxylation. The position of the methoxyl group appears to be important in the degradation of the aliphatic side chain of cinnamate; consequently, 3 methoxycinnamate and 4-methoxycinnamate, but not 2-methoxycinnamate, are transformed to produce acetate and methoxybenzoates, namely 3-methoxybenzoate and 4-methoxybenzoate, respectively. Crotonate is degraded to acetate and butyrate. The G+C content of the DNA is 56 mol%. Phylogenetic analysis of the 16S rRNA gene of strain CIN1T indicated that it was a member of the low-G+C-containing Gram positive branch with a specific relationship to Sporobacter termitidis (sequence identity of 88%). The phylogenetic results concur with the phenotypic data which reveals that the isolate is a novel bacterium and, based on these findings, strain CIN1T (= DSM 12816T = ATCC 700879T) has been designated Papillibacter cinnamivorans gen. nov., sp. nov. PMID- 10843067 TI - Halothiobacillus kellyi sp. nov., a mesophilic, obligately chemolithoautotrophic, sulfur-oxidizing bacterium isolated from a shallow-water hydrothermal vent in the Aegean Sea, and emended description of the genus Halothiobacillus. AB - A new mesophilic, chemolithoautotrophic, sulfur-oxidizing bacterium, strain Milos BII1T, was isolated from a sediment sample taken from a shallow-water hydrothermal vent in the Aegean Sea with thiosulfate as electron donor and CO2 as carbon source. Based on the almost complete sequence of the 16S rRNA gene, strain Milos-BII1T forms a phylogenetic cluster with Thiobacillus hydrothermalis, Thiobacillus neapolitanus, Thiobacillus halophilus and Thiobacillus sp. W5, all of which are obligately chemolithoautotrophic bacteria. Because of their phylogenetic relatedness and their physiological similarities it is proposed to transfer these organisms to a newly established genus within the gamma-subclass of the Proteobacteria, Halothiobacillus gen. nov. (Kelly and Wood 2000). Strain Milos-BII1T represents a new species of this genus, named Halothiobacillus kellyi. Cells were Gram-negative rods and highly motile. The organism was obligately autotrophic and strictly aerobic. Nitrate was not used as electron acceptor. Chemolithoautotrophic growth was observed with thiosulfate, tetrathionate, sulfur and sulfide. Growth was observed between pH values of 3.5 and 8.5, with an optimum at pH 6.5. The temperature limits for growth were 3.5 and 49 degrees C, with an optimum between 37 and 42 degrees C. Growth occurred between 0 and 2 M NaCl, with an optimum NaCl concentration between 400 and 500 mM. The mean maximum specific growth rate on thiosulfate was 0.45 h(-1). PMID- 10843068 TI - Desulfacinum hydrothermale sp. nov., a thermophilic, sulfate-reducing bacterium from geothermally heated sediments near Milos Island (Greece). AB - A thermophilic, sulfate-reducing bacterium, strain MT-96T, was isolated from an active, marine, shallow-water hydrothermal vent system. It used a large variety of substrates, ranging from simple organic compounds to long-chain fatty acids, as electron donors. Autotrophic growth was possible with H2 and CO2 in the presence of sulfate. Sulfate, thiosulfate and sulfite were used as electron acceptors. Sulfur and nitrate were not reduced. Fermentative growth was obtained with pyruvate, but not with fumarate or malate. Substrate oxidation was usually complete, leading to production of CO2, but at high substrate concentrations acetate accumulated. The oval-shaped cells were 0.8-1.0 microm in width and 1.5 2.5 microm in length. Cells were motile during the early-exponential-growth phase, but motility rapidly declined during later growth phases. Spores were not produced and cells stained Gram-negative. The temperature limits for growth were between 37 and 64 degrees C, with an optimum at 60 degrees C. Growth was observed at salinities ranging from 15 to 78 g NaCl l(-1), with optimum growth in the presence of 32-36 g NaCl l(-1). This might reflect an adaptation to the elevated salinity of the hydrothermal fluid. The G+C content of the DNA was 59.5 mol%. Vitamins or other supplements were not required. Based on the 16S rRNA gene sequence, strain MT-96T belonged in the delta-subclass of the Proteobacteria. Strain MT-96T was found to be phenotypically and phylogenetically related to Desulfacinum infernum (< 95.3% sequence identity) and represents a new member of the genus Desulfacinum. The name Desulfacinum hydrothermale is proposed for this strain; the type strain is MT-96T (= DSM 13146). PMID- 10843069 TI - Characterization of a Rothia-like organism from a mouse: description of Rothia nasimurium sp. nov. and reclassification of Stomatococcus mucilaginosus as Rothia mucilaginosa comb. nov. AB - An unknown, Gram-positive, ovoid-shaped bacterium isolated from the nose of a mouse was subjected to a polyphasic taxonomic analysis. Comparative 16S rRNA gene sequencing demonstrated that the unknown organism was a member of the family Micrococcaceae and possessed a specific phylogenetic association with Rothia dentocariosa and Stomatococcus mucilaginosus. Phenotypically, the bacterium closely resembled R. dentocariosa and S. mucilaginosus but could be distinguished from these species by biochemical tests and electrophoretic analysis of whole cell proteins. Based on both phylogenetic and phenotypic evidence, it is proposed that the unknown bacterium be classified in the genus Rothia, as Rothia nasimurium sp. nov. In addition, it is proposed that S. mucilaginosus be reclassified in the genus Rothia, as Rothia mucilaginosa comb. nov. PMID- 10843070 TI - Lactobacillus fornicalis sp. nov., isolated from the posterior fornix of the human vagina. AB - Twelve strains isolated from the posterior fornix fluid of the human vagina were identified as Lactobacillus johnsonii, Lactobacillus acidophilus, Lactobacillus gallinarum and Lactobacillus crispatus based on numerical analyses of total soluble cell protein profiles and randomly amplified polymorphic DNA (RAPD)-PCR banding patterns. Five strains grouped with the type strains of Lactobacillus gasseri (DSM 20077T) and Lactobacillus jensenii (DSM 20557T) at r > or = 0.83 in one protein profile cluster, well separated from the other species included in this study. However, numerical analysis of the RAPD-PCR banding patterns of representative strains selected from the L. gasseri-L. jensenii protein cluster clearly indicated that they belong to two different species. Four strains (TV 1010, TG 1013, TV 1018T and TV 1045) grouped into another well separated protein profile cluster at r > or = 0.87. Strains selected from this cluster displayed very similar RAPD-PCR banding patterns and clustered at R2 > or = 0.78, separate from the other strains examined. Sequencing of the 16S rRNA of two representative strains, TV 1018T and TG 1013, of this group indicated that it represents a new member of rRNA group I Lactobacillus, which includes Lactobacillus delbrueckii, the type of the genus, and close relatives Lactobacillus acetotolerans, Lactobacillus kefiranofaciens, Lactobacillus iners, L. jensenii, L. crispatus, L. acidophilus, Lactobacillus helveticus, Lactobacillus amylovorus, Lactobacillus hamsteri, L. johnsonii, L. gasseri and Lactobacillus amylolyticus. The name Lactobacillus fornicalis sp. nov. is proposed for strains TV 1010 (DSM 13172), TG 1013, TV 1018T and TV 1045, with strain TV 1018T (= DSM 13171T = ATCC 700934T) as the type. PMID- 10843071 TI - Clostridium peptidivorans sp. nov., a peptide-fermenting bacterium from an olive mill wastewater treatment digester. AB - A new peptide-degrading, strictly anaerobic bacterium, designated strain TMC4T, was isolated from an olive mill wastewater treatment digester. Cells of strain TMC4T were motile, rod-shaped (5-10 x 0.6-1.2 microm), stained Gram-positive and formed terminal to subterminal spores that distended the cells. Optimal growth occurred at 37 degrees C and pH 7 in an anaerobic basal medium containing 0.5% Casamino acids. Arginine, lysine, cysteine, methionine, histidine, serine, isoleucine, yeast extract, peptone, Biotrypcase, gelatin and crotonate also supported growth, but not carbohydrates, organic acids or alcohols. The end products of degradation were: acetate and butyrate from lysine and crotonate; acetate, butyrate, H2 and CO2 from Biotrypcase, gelatin and peptone; acetate, alanine, H2 and CO2 from cysteine; acetate, H2 and CO2 from serine, cysteine and yeast extract; acetate and formate from histidine; propionate from methionine; methyl 2-butyrate, H2 and CO2 from isoleucine; acetate and ethanol from arginine; and acetate, propionate, butyrate, methyl 2-butyrate, H2 and CO2 from Casamino acids. The DNA G+C content of strain TMC4T was 31 mol%. Phylogeny based on 16S rRNA sequence analysis showed that strain TMC4T was a member of the low-G+C content Gram-positive genus Clostridium, with the closest relative being Clostridium pascui (sequence similarity of 96 %). Due to considerable differences in genomic and phenotypic properties between strain TMC4T and those of its nearest relative, strain TMC4T is proposed as a new species of the genus Clostridium, Clostridium peptidivorans sp. nov. Strain TMC4T has been deposited in the DSMZ as strain DSM 12505T. PMID- 10843073 TI - Description of Bogoriellaceae fam. nov., Dermacoccaceae fam. nov., Rarobacteraceae fam. nov. and Sanguibacteraceae fam. nov. and emendation of some families of the suborder Micrococcineae. AB - The hierarchic taxonomic framework described recently for the phylogenetic structure of the suborder Micrococcineae, class Actinobacteria, on the basis of 16S rDNA sequences and signature nucleotides was modified and extended. With the recent addition of novel taxa into the suborder, the phylogenetic coherence of some families was disrupted, leading to the emergence of novel lineages that, as judged by the depth of their branching points, were equivalent to those of described families. Bogoriellaceae fam. nov., Dermacoccaceae fam. nov., Rarobacteraceae fam. nov. and Sanguibacteraceae fam. nov. are proposed for these lineages. As a consequence of the restructuring process, some families have had to be emended, i.e. Dermatophilaceae, Cellulomonadaceae and Intrasporangiaceae. PMID- 10843072 TI - The halotolerance and phylogeny of cyanobacteria with tightly coiled trichomes (Spirulina Turpin) and the description of Halospirulina tapeticola gen. nov., sp. nov. AB - The morphologies, halotolerances, temperature requirements, pigment compositions and 16S rRNA gene sequences of five culture collection strains and six novel isolates of cyanobacteria with helical, tightly coiled trichomes were investigated. All strains were very similar morphologically and could be assigned to the genus Spirulina (or section Euspirulina sensu Geitler), according to traditional classification. However, the isolates showed significantly different requirements for salinity and temperature, which were in accordance with their respective environmental origins. The genetic divergence among the strains investigated was large. The results indicate the drastic underestimation of the physiological and phylogenetic diversity of these cyanobacteria by the current morphology-based classification and the clear need for new taxa. Three of the isolates originated from hypersaline waters and were similar with respect to their high halotolerance, broad euryhalinity and elevated temperature tolerance. By phylogenetic analyses, they were placed in a tight monophyletic cluster apart from all other cyanobacteria. Thus it is proposed to reclassify highly halotolerant cyanobacteria with tightly coiled trichomes in Halospirulina gen. nov., with the type species Halospirulina tapeticola sp. nov. PMID- 10843074 TI - Thermanaerovibrio velox sp. nov., a new anaerobic, thermophilic, organotrophic bacterium that reduces elemental sulfur, and emended description of the genus Thermanaerovibrio. AB - A moderately thermophilic, organotrophic bacterium with vibrioid cells was isolated from a sample of a cyanobacterial mat from caldera Uzon, Kamchatka, Russia, and designated strain Z-9701T. Cells of strain Z-9701T were curved, Gram negative rods, 0.5-0.7 x 2.5-5.0 microm in size, with tapering ends and with fast, wavy movement by means of lateral flagella located on the concave side of the cell. Colonies were small, white, irregular or round, 0.2 mm in diameter, and with even edges. Strain Z-9701T was an obligate anaerobe with a temperature optimum at 60-65 degrees C and a pH optimum at 7.3. It fermented glucose, fructose, mannose, N-acetyl-D-glucosamine, adonite, arginine, serine, peptone, yeast extract and Casamino acids. The fermentation products formed during growth on glucose were acetate, lactate, H2, CO2 and ethanol. Strain Z-9701T reduced elemental sulfur to H2S during organotrophic growth with glucose or peptides as energy and carbon sources. In the presence of S0, strain Z-9701T was capable of lithotrophic growth with molecular hydrogen as energy substrate and 0.1 g yeast extract l(-1) as carbon source. Sulfate, thiosulfate, nitrate, Fe(III) and sulfite were not reduced and did not stimulate growth. The G+C content of strain Z-9701T DNA was 54.6 mol%. The results of 16S rDNA sequence analyses revealed that strain Z-9701T belongs to the cluster within the Clostridium group formed by Thermanaerovibrio acidaminovorans, Dethiosulfovibrio peptidovorans, Anaerobaculum thermoterrenum and Aminobacterium colombiense, but the level of sequence similarity with the members of this cluster was not very high (87.6-92.2%). Among these organisms, Thermanaerovibrio acidaminovorans is phenotypically close to strain Z-9701T. However, the two organisms showed a relatively low level of similarity of their 16S rRNA sequences (92.2%) and of DNA-DNA hybridization (15 +/- 1%). Nevertheless, on the basis of the similar morphology and physiology of the new isolate and Thermanaerovibrio acidaminovorans, strain Z-9701T was placed in the genus Thermanaerovibrio and a new species, Thermanaerovibrio velox, proposed for it. The type strain is Z-9701T (= DSM 12556T). PMID- 10843075 TI - Natrinema versiforme sp. nov., an extremely halophilic archaeon from Aibi salt lake, Xinjiang, China. AB - A novel extremely halophilic archaeon, strain XF10T, was isolated from a salt lake in China. This organism was neutrophilic, non-motile and pleomorphic, and was rod, coccus or irregularly shaped. It required at least 1.5 M NaCl for growth and grew in a wide range of MgCl2 concentrations (0.005-0.5 M). Lipid extract of whole cells contained two glycolipids with the same chromatographic properties as two unidentified glycolipids found in the two described Natrinema species, Natrinema pellirubrum and Natrinema pallidum. Phylogenetic analysis based on 16S rDNA sequence comparison revealed that strain XF10T clustered with the two described Natrinema species and several other strains (strains T5.7, GSL-11 and Haloterrigena turkmenica JCM 9743) with more than 98.1% sequence similarities, suggesting that strain XF1OT belongs to the genus Natrinema. Comparative analysis of phenotypic properties and DNA-DNA hybridization between strain XF10T and the Natrinema species supported the conclusion that strain XF10T is a novel species within the genus Natrinema. The name Natrinema versiforme sp. nov. is proposed for this strain. The type strain is XF10T (=JCM 10478T=AS 1.2365T=ANMR 0149T). PMID- 10843076 TI - New psychrophilic and psychrotolerant Bacillus marinus strains from tropical and polar deep-sea sediments and emended description of the species. AB - In contrast to the current view that psychrophily combined with an absolute requirement for NaCl is connected with the Gram-negative cell wall type, psychrophilic and psychrotolerant, NaCl-requiring, Gram-positive bacteria have been isolated from tropical Atlantic, Arctic and Antarctic deep-sea sediments. Some of the isolates are even extremely psychrophilic, having maximum growth temperatures of 4 degrees C. On the basis of phenotypic characteristics, DNA base analyses, DNA-DNA hybridizations and partial and complete 16S rRNA gene sequence analyses, the strains from the three distinct geographical regions have been allocated to the obligately marine species Bacillus marinus. The distribution and origin of B. marinus are discussed and an emended description of the species is presented. PMID- 10843077 TI - Saccharothrix violacea sp. nov., isolated from a gold mine cave, and Saccharothrix albidocapillata comb. nov. AB - The generic position of two isolates from soils inside a gold mine cave in Kongju, Korea, was determined by 16S rDNA sequencing and chemotaxonomic characteristics. Phylogenetic analysis indicated that both of the isolates formed a clade with Lentzea albidocapillata and members of the genus Saccharothrix of the family Pseudonocardiaceae. The chemical composition of the isolates and of Lentzea albidocapillata was consistent with that of the genus Saccharothrix, which is characterized by a type III cell wall (the meso-isomer of diaminopimelic acid, and galactose and rhamnose as characteristic whole-cell sugars), MK-9(H4) as the major menaquinone, and a phospholipid type PII pattern (phosphatidylethanolamine as a diagnostic phospholipid). The combination of morphological features, chemotaxonomic characters and phylogenetic data supported the proposal that Lentzea albidocapillata, the only and type strain of the genus Lentzea, should be transferred to the genus Saccharothrix. On the basis of physiological properties, cellular fatty acid composition and DNA-DNA hybridization data, two new species within the genus Saccharothrix are proposed: Saccharothrix violacea sp. nov., type strain LM 036T (= IMSNU 50388T), and Saccharothrix albidocapillata comb. nov., type strain DSM 44073T (=IMSNU 21253T). PMID- 10843078 TI - Comparison of the 16S-23S rRNA intergenic spacer regions among strains of the Mycoplasma mycoides cluster, and reassessment of the taxonomic position of Mycoplasma sp. bovine group 7. AB - Nucleotide sequence analysis of the 16S-23S rRNA intergenic spacer regions of six type or reference strains belonging to the Mycoplasma mycoides cluster and of Mycoplasma putrefaciens suggested the presence of two subclusters. One subcluster comprised M. mycoides subsp. mycoides small colony (SC) type, M. mycoides subsp. mycoides large colony (LC) type and M. mycoides subsp. capri, whereas the second subcluster comprised Mycoplasma capricolum subsp. capricolum, M. capricolum subsp. capripneumoniae and Mycoplasma sp. bovine group 7. The type strains from M. mycoides subsp. mycoides SC and M. mycoides subsp. capri had identical spacer sequences. The existence of two subclusters was supported by predicted secondary structures of the analysed region. The nucleotide variations in the loop domains of the secondary structures might be a useful genetic marker to distinguish between the two subclusters. The secondary structure differences delineated the differences between the two subclusters more clearly than the nucleotide sequence alignments, which only showed a small number of differences, and some of these were common to both clusters. The data also provided evidence in favour of a reclassification of Mycoplasma sp. bovine group 7 as another subspecies of M. capricolum. PMID- 10843079 TI - Bacillus thermodenitrificans sp. nov., nom. rev. AB - A polyphasic study was performed on 10 soil isolates of thermophilic denitrifying Bacillus strains from different geographical areas. The presence of two main characteristic bands following amplification of the internal transcribed spacer (ITS) region of rrn operons suggests a close relatedness to 'Bacillus thermodenitrificans'. The isolates cluster around two strains of 'B. thermodenitrificans' in riboprint and fatty acid analyses, though differences occur at the strain level. Subsequent DNA-DNA reassociation studies including the 10 isolates, 'B. thermodenitrificans' DSM 465T and DSM 466, and Bacillus stearothermophilus ATCC 12980T and Bacillus thermoleovorans ATCC 43513T revealed such a high level of genomic relatedness between the isolates and the DSM strains (> 73% similarity) that they must be considered strains of the same taxon. The degree of DNA-DNA similarity between the 12 strains of 'B. thermodenitrificans' and the type strains of the other two phylogenetically neighbouring Bacillus species was significantly lower (21-43% similarity). Based upon phylogenetic, chemotaxonomic and phenotypic evidence, the designation of B. thermodenitrificans sp. nov., nom. rev. is proposed. The type strain of B. thermodenitrificans is DSM 465T. PMID- 10843080 TI - Photobacterium histaminum Okuzumi et al. 1994 is a later subjective synonym of Photobacterium damselae subsp. damselae (Love et al. 1981) Smith et al. 1991. AB - The type strain of Photobacterium histaminum, JCM 8968T (= ATCC 51805T), and that of Photobacterium damselae subsp. damselae, ATCC 33539T, exhibit 100% identity in their 16S rRNA sequence, more than 80% DNA-DNA homology and only one phenotypic difference. Also, like P. histaminum, P. damselae subsp. damselae was shown to excrete a large amount of histamine when cells were grown on medium containing excessive histidine under acidic conditions. Therefore, the name P. histaminum should be considered to be a later subjective synonym of P. damselae subsp. damselae. PMID- 10843081 TI - Grouping oral Candida species by multilocus enzyme electrophoresis. AB - Multilocus enzyme electrophoresis (MLEE) and numerical taxonomic methods were used to establish the degrees of relatedness among five Candida species commonly isolated from humans oral cavities. Of twenty enzymic systems assayed, five showed no enzymic activity (aspartate dehydrogenase, mannitol dehydrogenase, sorbitol dehydrogenase, glucosyl transferase and alpha-amylase). The obtained data revealed that some of these enzymes are capable of distinguishing strains of different species, but most of them could not organize all strains in their respective species-specific clusters. Numerical classification based on MLEE polymorphism must be regarded for surveys involving just one Candida species. PMID- 10843082 TI - Biodiversity and systematics of basidiomycetous yeasts as determined by large subunit rDNA D1/D2 domain sequence analysis. AB - The molecular systematics of 337 strains of basidiomycetous yeasts and yeast-like fungi, representing 230 species in 18 anamorphic and 24 teleomorphic genera, was determined by sequence analysis of the D1/D2 region of the large-subunit rDNA. The data were compared with published sequences of other basidiomycetous fungi. The results demonstrated that the yeast species and genera are phylogenetically distributed among the Microbotryum, Sporidiobolus, Agaricostilbum and Erythrobasidium clades of the Urediniomycetes; the Tremellales, Trichosporonales ord. nov., Filobasidiales and Cystofilobasidiales clades of the Hymenomycetes; and the Ustilaginales, Microstromatales and Malasseziales clades of the Ustilaginomycetes. Genera such as Bensingtonia, Cryptococcus, Rhodotorula and Sporobolomyces are polyphyletic, i.e. they occur in two or more clades. In contrast, other genera, e.g. Bullera, Cystofilobasidium, Fellomyces, Filobasidiella, Filobasidium, Kondoa, Kurtzmanomyces, Leucosporidium, Rhodosporidium, Sporidiobolus and Udeniomyces, are monophyletic. The majority of the species can be identified using D1/D2 analyses, although the internal transcribed spacer region is required to distinguish closely related species. The intergenic spacer region is recommended for additional differentiation of species and strains. PMID- 10843083 TI - Phylogenetic analysis of the ballistoconidium-forming yeast genus Sporobolomyces based on 18S rDNA sequences. AB - The 18S rDNA nucleotide sequences of 25 Sporobolomyces species and five Sporidiobolus species were determined. Those of Sporobolomyces dimmenae JCM 8762T, Sporobolomyces ruber JCM 6884T, Sporobolomyces sasicola JCM 5979T and Sporobolomyces taupoensis JCM 8770T showed the presence of intron-like regions with lengths of 1586, 324, 322 and 293 nucleotides, respectively, which were presumed to be group I introns. A total of 63 18S rDNA nucleotide sequences was analysed, including 33 published reference sequences. Sporobolomyces species and the other basidiomycetes species were distributed throughout the phylogenetic tree. The resulting phylogeny indicated that Sporobolomyces is polyphyletic. Sporobolomyces species were mainly divided into four groups within the Urediniomycetes. The groups are designated as the Sporidiales, Agaricostilbum/Bensingtonia, Erythrobasidium and subbrunneus clusters. The last group, comprising four species, Sporobolomyces coprosmicola, Sporobolomyces dimmenae, Sporobolomyces linderae and Sporobolomyces subbrunneus, forms a new and distinct cluster in the phylogenetic tree in this study. PMID- 10843085 TI - International Committee on Systematic Bacteriology Subcommittee on the taxonomy of Bifidobacterium, Lactobacillus and related organisms. Minutes of the meetings, 4 and 6 July 1994, Prague, Czech Republic. PMID- 10843084 TI - DNA fingerprinting patterns of Candida species using HinfI endonuclease. AB - Strain delineation was performed by means of restriction endonuclease analysis (REA) of genomic DNA with the restriction enzyme HinfI followed by conventional electrophoresis. A total of 337 yeast isolates representing 21 Candida species and five non-Candida yeast species was evaluated. A survey of isolates showed that Candida albicans and non-albicans species could be divided into mutually exclusive groups, and that subgroups could be created. Individual REA patterns for 111 C. albicans isolates, four Candida krusei isolates and 35 Candida glabrata isolates varied greatly, whereas 11 Candida dubliniensis isolates, 48 Candida tropicalis isolates and 41 Candida guilliermondii isolates could be divided into two, nine and nine groups, respectively. REA of the 49 Candida parapsilosis isolates with HinfI, however, showed that 47 (95 %) of them belonged to one group. REA patterns of the other yeast isolates, representing 19 species, were also quite different at the species level. These results showed that REA with HinfI may be useful for the identification and strain delineation of common and emerging Candida species. PMID- 10843086 TI - International Committee on Systematic Bacteriology Subcommittee on the taxonomy of Mollicutes. Minutes of the meetings, 21 and 28 July 1998, Sydney, Australia. PMID- 10843087 TI - International Committee on Systematic Bacteriology Subcommittee on the taxonomy of Campylobacter and related bacteria. Minutes of the meeting, 2 September 1998, Budapest, Hungary. PMID- 10843088 TI - International Committee on Systematic Bacteriology Subcommittee on the taxonomy of Campylobacter and related bacteria. Minutes of the meetings, 13 and 16 September 1999, Baltimore, USA. PMID- 10843089 TI - International Committee on Systematic Bacteriology Subcommittee on the taxonomy of Halobacteriaceae. Minutes of the meetings, 16 August 1999, Sydney, Australia. PMID- 10843090 TI - International Committee on Systematic Bacteriology Subcommittee on the taxonomy of genus Bacillus and related organisms. Minutes of the meeting, 18 August 1999, Sydney, Australia. PMID- 10843091 TI - Hyperthermia treatment planning. AB - The development of hyperthermia, the treatment of tumours with elevated temperatures in the range of 40-44 degrees C with treatment times over 30 min, greatly benefits from the development of hyperthermia treatment planning. This review briefly describes the state of the art in hyperthermia technology, followed by an overview of the developments in hyperthermia treatment planning. It particularly highlights the significant problems encountered with heating realistic tissue volumes and shows how treatment planning can help in designing better heating technology. Hyperthermia treatment planning will ultimately provide information about the actual temperature distributions obtained and thus the tumour control probabilities to be expected. This will improve our understanding of the present clinical results of thermoradiotherapy and thermochemotherapy, and will greatly help both in optimizing clinical heating technology and in designing optimal clinical trials. PMID- 10843092 TI - A semi-empirical treatment planning model for optimization of multiprobe cryosurgery. AB - A model is presented for treatment planning of multiprobe cryosurgery. In this model a thermal simulation algorithm is used to generate temperature distribution from cryoprobes, visualize isotherms in the anatomical region of interest (ROI) and provide tools to assist estimation of the amount of freezing damage to the target and surrounding normal structures. Calculations may be performed for any given freezing time for the selected set of operation parameters. The thermal simulation is based on solving the transient heat conduction equation using finite element methods for a multiprobe geometry. As an example, a semi-empirical optimization of 2D placement of six cryoprobes and their thermal protocol for the first freeze cycle is presented. The effectiveness of the optimized treatment protocol was estimated by generating temperature-volume histograms and calculating the objective function for the anatomy of interest. Two phantom experiments were performed to verify isotherm locations predicted by calculations. A comparison of the predicted 0 degrees C isotherm with the actual iceball boundary imaged by x-ray CT demonstrated a spatial agreement within +/-2 mm. PMID- 10843093 TI - Dynamics of pulsed holmium:YAG laser photocoagulation of albumen. AB - The pulsed holmium:YAG laser (lambda = 2.12 microm, tau(p) = 250 micros) has been investigated as a method for inducing localized coagulation for medical procedures, yet the dynamics of this process are not well understood. In this study, photocoagulation of albumen (egg white) was analysed experimentally and results compared with optical-thermal simulations to investigate a rate process approach to thermal damage and the role of heat conduction and dynamic changes in absorption. The coagulation threshold was determined using probit analysis, and coagulum dynamics were documented with fast flash photography. The nonlinear computational model, which included a Beer's law optical component, a finite difference heat transfer component and an Arrhenius equation-based damage calculation, was verified against data from the literature. Moderate discrepancies between simulation results and our experimental data probably resulted from the use of a laser beam with an irregular spatial profile. This profile produced a lower than expected coagulation threshold and an irregular damage distribution within a millisecond after laser onset. After 1 ms, heat conduction led to smoothing of the coagulum. Simulations indicated that dynamic changes in absorption led to a reduction in surface temperatures. The Arrhenius equation was shown to be effective for simulating transient albumen coagulation during pulsed holmium:YAG laser irradiation. Greater understanding of pulsed laser-tissue interactions may lead to improved treatment outcome and optimization of laser parameters for a variety of medical procedures. PMID- 10843094 TI - Dynamic contrast enhanced CT measurement of blood flow during interstitial laser photocoagulation: comparison with an Arrhenius damage model. AB - One effect of heating during interstitial laser photocoagulation (ILP) is to directly destroy the tumour vasculature resulting in a loss of viable blood supply. Therefore, blood flow measured during and after treatment can be a useful indicator of tissue thermal damage. In this study, the effect of ILP treatment on rabbit thigh tumours was investigated by measuring blood flow changes using dynamic contrast enhanced computed tomography (CT). The CT measured changes in blood flow of treated tumour tissue were fitted to an Arrhenius model assuming first order rate kinetics. Our results show that changes in blood flow of tumour tissue distant from surrounding normal tissue are well described by an Arrhenius model. By contrast, the temperature profile of tumour tissue adjacent to normal tissue must be modified to account for heat dissipation by the latter. Finally, the Arrhenius parameters derived in the study are similar to those derived by heating tumour tissue to a lower temperature (<47 degrees C) than the current study. In conclusion, CT can be used to monitor blood flow changes during ILP and these measurements are related to the thermal damage predicted by the Arrhenius model. PMID- 10843095 TI - High-angle scattering events strongly affect light collection in clinically relevant measurement geometries for light transport through tissue. AB - Measurement of light transport in tissue has the potential to be an inexpensive and practical tool for non-invasive tissue diagnosis in medical applications because it can provide information on both morphological and biochemical properties. To capitalize on the potential of light transport as a diagnostic tool, an understanding of what information can be gleaned from light transport measurements is needed. We present data concerning the sensitivity of light transport measurements, made in clinically relevant geometries, to scattering properties. The intensity of the backscattered light at small source-detector separations is shown to be sensitive to the phase function, and furthermore the collected light intensity is found to be correlated with the amount of high-angle scattering in the medium. PMID- 10843096 TI - Effect of myocardial anisotropy on the torso current flow patterns, potentials and magnetic fields. AB - The effects of myocardial anisotropy on the torso current flow patterns, voltage and the magnetic field were examined using an anatomically realistic torso model of an adult male subject. A finite element model of the torso was built with 19 major tissue types identified. The myocardial fibre orientation in the heart wall was included with a voxel resolution of 0.078 x 0.078 x 0.3 cm. The fibre orientations from the canine heart which are available in the literature were mapped to our adult male subject's human heart using deformable mapping techniques. The current and potential distribution in the whole torso were computed using an idealized dipolar source of +/-1.0 V in the middle of the septum of the heart wall as a boundary condition. An adaptive finite element solver was used. Two cases were studied. In one case the myocardium was isotropic and in the other it was anisotropic. It was found that the current density distribution shows a very noticeable difference between the isotropic and anisotropic myocardium. The resultant magnetic field in front of the torso was computed using the Biot-Savart law. It was found that the magnetic field profile was slightly affected by the myocardial anisotropy. The potential on the torso surface also shows noticeable changes due to the myocardial anisotropy. PMID- 10843097 TI - Development of a compact proton scanning system in Uppsala with a moveable second magnet. AB - A scanned proton beam yields dose distributions that in most cases are superior to passively scattered proton beams and to other external radiation treatment modalities. The present paper gives a description of the scanning system that has been developed at the Svedberg Laboratory (TSL) in Uppsala. The scanning technique and the technical design are described. The solution with a small pole gap of the magnets and a moveable second magnet results in a very compact scanning head, which can therefore be incorporated in a gantry of relatively limited size. A prototype was constructed that has been used to realize various dose distributions with a scanned beam of 180 MeV protons at TSL. PMID- 10843098 TI - Dealing with Cerenkov radiation generated in organic scintillator dosimeters by bremsstrahlung beams. AB - An organic scintillator detector system has been developed for radiotherapy bremsstrahlung dosimetry. The scintillators are connected to photodiodes by light pipes as the photodiodes must be removed and shielded from the incident radiation. The photodiodes see visible and near-visible light emissions from the scintillator as well as Cerenkov and fluorescence radiation that has been generated and trapped in the scintillator and light pipe. The Cerenkov and fluorescence radiation limits the accuracy of the dosimeter. This work examines a range of methods for diminishing the signal contribution of Cerenkov and fluorescence radiation while optimizing the scintillator signal. Three methods of achieving these goals have been used. They are: reflective coatings on the scintillator, long-wavelength-emitting scintillators used in conjunction with the photodiode, and absorptive filters placed between the light guide and photodiode. The contribution of the Cerenkov radiation to the light seen by the photodiode has been modelled and the model predictions have been tested using bremsstrahlung beams of peak energy between 13 and 20 MV, showing agreement with measurement. PMID- 10843099 TI - Methods for separation of contributions from two radionuclides in autoradiography with a silicon strip detector. AB - Two methods have been developed for the separation of contributions from two different nuclides in an autoradiography picture. Both approaches, a modified least squares (LS) and a maximum likelihood (ML) algorithm, are based on the position and energy information available from a digital detector. Tests and comparisons, using artificially as well as measured data, demonstrate that the ML approach performs slightly better, but is much more computationally demanding than the LS method. PMID- 10843100 TI - The reproducibility of polyacrylamide gel dosimetry applied to stereotactic conformal radiotherapy. AB - The reproducibility of polyacrylamide gel (PAG) dosimetry has been evaluated when used to verify two radiotherapy treatment plans of increasing complexity. The plans investigated were a three-field coplanar arrangement, using the linac jaws for field shaping, and a four-field, conformal, non-coplanar plan using precision cast lead alloy shielding blocks. Each treatment was performed three times using phantoms and calibration gels manufactured in-house. Two phantoms were specially designed for this work to aid accurate positioning of the gels for irradiation and imaging. All gels were imaged post-irradiation using a Siemens Vision 1.5T MR scanner. T2 relaxation images were calibrated to absorbed dose distributions using a number of smaller calibration vessels to produce distribution maps of relative dose. The relative dose distributions were found to be reproducible, with the standard deviation on the mean areas enclosed by the > or = 50% isodose lines measured in three orthogonal planes being 6.4% and 4.1% for the coplanar and non-coplanar plans respectively. The measured distributions were also consistent with those planned, with isodose lines generally agreeing to within a few millimetres. However, the measured absolute doses were on average 23.5% higher than those planned. Although the polyacrylamide gel dosimetry technique has some limitations, particularly when calibrating distributions to absolute dose, the ability to resolve sharp dose gradients in three dimensions with millimetre precision is invaluable when verifying complex conformal treatment plans, where avoidance of proximal, critical structures is a treatment criterion. PMID- 10843101 TI - Requirements for leaf position accuracy for dynamic multileaf collimation. AB - Intensity modulated radiation therapy can be achieved by driving the leaves of a multileaf collimator (MLC) across an x-ray therapy beam. Algorithms to generate the required leaf trajectories assume that the leaf positions are exactly known to the MLC controller. In practice, leaf positions depend upon calibration accuracy and stability and may vary within set tolerances. The purpose of this study was to determine the effects of potential leaf position inaccuracies on intensity modulated beams. Equations are derived which quantify the absolute error in delivered monitor units given a known error in leaf position. The equations have been verified by ionization chamber measurements in dynamically delivered flat fields, comparing deliveries in which known displacements have been applied to the defined leaf positions with deliveries without displacements applied. The equations are then applied to two clinical intensity modulations: an inverse planned prostate field and a breast compensating field. It is shown that leaf position accuracy is more critical for a highly modulated low-dose intensity profile than a moderately modulated high-dose intensity profile. Suggestions are given regarding the implications for quality control of dynamic MLC treatments. PMID- 10843102 TI - Selection of beam angles for radiotherapy of skull base tumours using charged particles. AB - The geometrical treatment configuration for skull base tumours is investigated using three different sources of information. First, an analysis of treatment angle combinations for 50 patients treated with conventional conformal radiation therapy is performed. In a second step, nine treatment plans for a hypothetical heavy ion gantry were prepared. Finally, cylindrical projections of the organs at risk are introduced as a tool to analyse the distribution of treatment angles as well as the treatment geometry for each patient. The results of the analysis for conventional therapy clearly show treatment angle combinations that are preferably used. These findings are also supported by the geometrical analysis using the cylindrical projections. The majority of treatment angles for heavy ions also seem to be confined to those regions. In one case, however, the sharp distal dose fall-off of the heavy ions allowed a beam direction outside these conventionally used regions. The advantages of a heavy ion gantry versus a fixed horizontal beam line are shown by comparison of treatment plans for both systems. Treatment angle combinations are suggested that should be accessible for any beam line design using either fixed beams or a gantry. PMID- 10843103 TI - Multispectral imaging approach in the diagnosis of cutaneous melanoma: potentiality and limits. AB - In an attempt to overcome the subjectiveness of clinical observation in the diagnosis of cutaneous melanoma, a computerized method is proposed. Reflectance images of 237 pigmented lesions (67 melanomas and 170 non-melanomas) were analysed using a telespectrophotometric technique. This device consists of a CCD camera with 17 interference filters. Images were acquired at selected wavelengths, from 420 to 1040 nm. Morphological and reflectance related parameters were extracted from the wavelength-dependent images of the lesions. The most significant features in the comparison between benign and malignant lesions were: lesion dimension (P < 10(-8) at 578 nm); mean value (P < 10(-7) at 940 nm) and standard deviation (P < 10(-4) at 904 nm) of lesion reflectance; lesion roundness (P < 10(-5) at 461 nm); and border irregularity (P < 10(-4) at 461 nm). Based on these parameters, a discriminant function between the two populations of lesions (naevi and melanomas) was obtained. By using the results of the analysis of the recruited lesions as 'training data', discriminant functions enabled the assignment of a score, or a 'risk probability', to each studied lesion. By imposing a sensitivity of 80% (a figure that mimics the diagnostic capability of an experienced clinician), entering or not entering the lesion dimension as input data in the discriminant analysis led to a specificity of 51% or 46% respectively. The high number of false-positive cases, which is a consequence of the selection criteria of the lesions, is, at present, the major limitation of the current technique. Nevertheless, our results suggest that an imaging-based computer-assisted device could be capable of discriminating malignant lesions mainly by evaluation of reflectance, especially in the infrared region, and shape properties. The dimension of a lesion should not be essential in the diagnosis of melanoma and, in our opinion, small melanomas should be recognized by a computer system as well as they are on clinical grounds. PMID- 10843104 TI - Three-dimensional echocardiography: assessment of inter- and intra-operator variability and accuracy in the measurement of left ventricular cavity volume and myocardial mass. AB - Accurate left ventricular (LV) volume and mass estimation is a strong predictor of cardiovascular morbidity and mortality. We propose that our technique of 3D echocardiography provides an accurate quantification of LV volume and mass by the reconstruction of 2D images into 3D volumes, thus avoiding the need for geometric assumptions. We compared the accuracy and variability in LV volume and mass measurement using 3D echocardiography with 2D echocardiography, using in vitro studies. Six operators measured the LV volume and mass of seven porcine hearts, using both 3D and 2D techniques. Regression analysis was used to test the accuracy of results and an ANOVA test was used to compute variability in measurement. LV volume measurement accuracy was 9.8% (3D) and 18.4% (2D); LV mass measurement accuracy was 5% (3D) and 9.2% (2D). Variability in LV volume quantification with 3D echocardiography was %SEMinter = 13.5%, %SEMintra = 11.4%, and for 2D echocardiography was %SEMinter = 21.5%, %SEMintra = 19.1%. We derived an equation to predict uncertainty in measurement of LV volume and mass using 3D echocardiography, the results of which agreed with our experimental results to within 13%. 3D echocardiography provided twice the accuracy for LV volume and mass measurement and half the variability for LV volume measurement as compared with 2D echocardiography. PMID- 10843105 TI - A slice-by-slice blurring model and kernel evaluation using the Klein-Nishina formula for 3D scatter compensation in parallel and converging beam SPECT. AB - Converging collimation increases the geometric efficiency for imaging small organs, such as the heart, but also increases the difficulty of correcting for the physical effects of attenuation, geometric response and scatter in SPECT. In this paper, 3D first-order Compton scatter in non-uniform scattering media is modelled by using an efficient slice by-slice incremental blurring technique in both parallel and converging beam SPECT. The scatter projections are generated by first forming an effective scatter source image (ESSI), then forward-projecting the ESSI. The Compton scatter cross section described by the Klein-Nishina formula is used to obtain spatial scatter response functions (SSRFs) of scattering slices which are parallel to the detector surface. Two SSRFs of neighbouring scattering slices are used to compute two small orthogonal 1D blurring kernels used for the incremental blurring from the slice which is further from the detector surface to the slice which is closer to the detector surface. First-order Compton scatter point response functions (SPRFs) obtained using the proposed model agree well with those of Monte Carlo (MC) simulations for both parallel and fan beam SPECT. Image reconstruction in fan beam SPECT MC simulation studies shows increased left ventricle myocardium-to-chamber contrast (LV contrast) and slightly improved image resolution when performing scatter compensation using the proposed model. Physical torso phantom fan beam SPECT experiments show increased myocardial uniformity and image resolution as well as increased LV contrast. The proposed method efficiently models the 3D first-order Compton scatter effect in parallel and converging beam SPECT. PMID- 10843106 TI - Monte Carlo simulation of x-ray spectra in mammography. AB - A model for generating x-ray spectra in mammography is presented. This model used the ITS version 3 Monte Carlo code for simulating the radiation transport. Various target/filter combinations such as tungsten/aluminium, molybdenum/molybdenum, molybdenum/rhodium and rhodium/rhodium were used in the simulation. Both bremsstrahlung and characteristic x-ray production were included in the model. The simulated x-ray emission spectra were compared with two sets of spectra, those of Boone et al (1997 Med. Phys. 24 1863-74) and IPEM report 78. The chi2 test was used for the overall goodness of fit of the spectral data. There is good agreement between the simulated x-ray spectra and the comparison spectra as the test yielded a probability value of nearly 1. When the transmitted x-ray spectra for specific target/filter combinations were generated and compared with a measured molybdenum/rhodium spectrum and spectra generated in IPEM report 78, close agreement is also observed. This was demonstrated by the probability value for the chi2 test being almost 1 for all the cases. However, minor differences between the simulated spectra and the 'standard' ones are observed. PMID- 10843107 TI - Modelling of a 188W/188Re beta line source for coronary brachytherapy by means of EGS4 Monte Carlo simulations. AB - In this paper, we present results from three different simulation models that are used to determine the dose distribution around a 188W/188Re coronary brachytherapy source with EGS4 Monte Carlo simulations. The three models are found to give similar results within 10%. Agreement was found with experimental data from measurements in a PMMA phantom. It has been shown that in the therapeutically relevant region the beta line source can be characterized by the radial depth-dose distribution in water. PMID- 10843108 TI - The effects of dynamic optical properties during interstitial laser photocoagulation. AB - A nonlinear mathematical model was developed and experimentally validated to investigate the effects of changes in optical properties during interstitial laser photocoagulation (ILP). The effects of dynamic optical properties were calculated using the Arrhenius damage model, resulting in a nonlinear optothermal response. This response was experimentally validated by measuring the temperature rise in albumen and polyacrylamide phantoms. A theoretical study of ILP in liver was conducted constraining the peak temperatures below the vaporization threshold. The temperature predictions varied considerably between the static and dynamic scenarios, and were confirmed experimentally in phantoms. This suggests that the Arrhenius model can be used to predict dynamic changes in optical and thermal fields. An increase in temperature rise due to a decrease in light penetration within the coagulated region during ILP of the liver was also demonstrated. The kinetics of ILP are complex and nonlinear due to coagulation, which changes the tissue properties during treatment. These complex effects can be adequately modelled using an Arrhenius damage formulation. PMID- 10843110 TI - Changes in optical properties of ex vivo rat prostate due to heating. AB - This study examines the effectiveness of a single, first-order Arrhenius process in accurately modelling the thermally induced changes in the optical properties, particularly the reduced scattering coefficient, mu(s)', and the absorption coefficient, mu(a), of ex vivo rat prostate. Recent work has shown that mu(s)' can increase as much as five-fold due to thermal coagulation, and the observed change in mu(s)' has been modelled well according to a first-order rate process in albumen. Conversely, optical property measurements conducted using pig liver suggest that this change in mu(s)' cannot suitably be described using a single rate parameter. In canine prostate, measurements have indicated that while the absorption coefficient varies with temperature, it does not do so according to first-order kinetics. A double integrating sphere system was used to measure the reflectance and transmittance of light at 810 nm through a thin sample of prostate. Using prostate samples collected from Sprague Dawley rats, optical properties were measured at a constant elevated temperature. Tissue samples were measured over the range 54-83 degrees C. The optical properties of the sample were determined through comparison with reflectance and transmittance values predicted by a Monte Carlo simulation of light propagation in turbid media. A first order Arrhenius model was applied to the observed change in mu(s)' and mu(a) to determine the rate process parameters for thermal coagulation. The measured rate coefficients were Ea = (7.18 +/- 1.74) x 10(4) J mol(-1) and Afreq = 3.14 x 10(8) s(-1) for mu(s)'. It was determined that the change in mu(s)' is well described by a single first-order rate process. Similar analysis performed on the changes in mu(a) due to increased temperatures yielded Ea = (1.01 +/- 0.35) x 10(5) J mol(-1) and Afreq = 8.92 x 10(12) s(-1). The results for mu(a) suggest that the Arrhenius model may be applicable to the changes in absorption. PMID- 10843109 TI - Accuracy of the diffusion equation to describe photon migration through an infinite medium: numerical and experimental investigation. AB - The accuracy of results obtained from the diffusion equation (DE) has been investigated for the case of an isotropic point source in a homogeneous, weakly absorbing, infinite medium. The results from the DE have been compared both with numerical solutions of the radiative transfer equation obtained with Monte Carlo (MC) simulations and with cw experimental results. Comparisons showed that for the cw fluence rate, discrepancies are of the same order as statistical fluctuations on MC results (within 1%) when the distance r from the source is > 2/mu(s)', (mu(s)' is the reduced scattering coefficient). For these values of r, discrepancies for the time-resolved fluence rate are of the same order of statistical fluctuations (within 5%) when the time of flight is t > 4t0 with to time of flight for unscattered photons. For shorter times the DE overestimates the fluence discrepancies are larger for larger values of the asymmetry factor. As to the specific intensity, for small values of r the MC results are more forward peaked than expected from the DE, and the forward peak is stronger for photons arriving at short times. We assumed r > 2/mu(s)' and t > 4t0 for the domain of validity of the DE and we determined the requirements for which the simplifying assumptions necessary to obtain the DE, expressed by two inequalities, are fulfilled. Comparisons with cw experimental results showed a good agreement with MC results both at high and at small values of r mu(s)', while the comparison with the DE showed significant discrepancies for small values of r mu(s)'. Using MC results we also investigated the error made on the optical properties of the medium when they are retrieved using the solution of the DE. To obtain accuracy better than 1% from fitting procedures on time resolved fluence rate data it is necessary to disregard photons with time of flight < 4t0. Also from cw data it is possible to retrieve the optical properties with good accuracy: by using the added absorber technique discrepancies are < 1%, both on mu(s)' and on mu(a), if the absorption coefficient is small (mu(a)/mu(s)' < 0.005). PMID- 10843111 TI - The use of skin Fe levels as a surrogate marker for organ Fe levels, to monitor treatment in cases of iron overload. AB - A system based on the detection of K-shell x-ray fluorescence (XRF) has been used to investigate whether a correlation exists between the concentration of iron in the skin and the concentration of iron in the liver, as the degree of iron loading increases. The motivation behind this work is to develop a non-invasive method of determining the extent of the body's iron stores via measurements on the skin, in order to monitor the efficacy of chelation therapy administered to patients with beta-thalassaemia. Sprague-Dawley rats were iron loaded via injections of iron dextran and subsequently treated with the iron chelator CP94. The non-haem iron concentrations of the liver, heart and spleen were determined using bathophenanthroline sulphonate as the chromogen reagent. Samples of abdominal skin were taken and the iron concentrations determined using XRF. A strong correlation between the skin iron concentration and the liver iron concentration has been demonstrated (R2 = 0.86). Similar correlations exist for the heart and the spleen. These results show that this method holds great potential as a tool in the diagnosis and treatment of hereditary haemochromatosis and beta-thalassaemia. PMID- 10843112 TI - Multifrequency method for dielectric monitoring of cold-preserved organs. AB - To answer a growing need for non-invasive monitoring of biological organs, we have developed an automated system capable of repeated dielectric measurements over the frequency range 10 kHz-100 MHz. Further, we propose a novel method of data analysis that may convert the acquired, individual dispersion curves into a diagram of the time course of specific phenomenological parameters, such as the characteristic frequency. By using this new procedure, unattended, long-term monitoring of temporal changes in the dielectric behaviour of excised liver lobes stored at 4 degrees C was successfully realized. The 'multifrequency' method presented here was definitely superior to the conventional 'fixed-frequency' method in providing reliable results. PMID- 10843113 TI - Isocentric treatment of inclined volumes planned using coronal sections. AB - The treatment parameters necessary for the isocentric treatment of an inclined volume have to be determined either analytically or through simulation. The derivation of the treatment parameters for the treatment of a transverse plane has been described previously. This work describes the derivation of the treatment parameters necessary for the isocentric treatment of an inclined volume that has been planned from an angled coronal section. Ways of implementing the system in the clinic are described. PMID- 10843114 TI - The effect of user-defined variables on dosimetry consistency in Gamma Knife planning. AB - We report a dosimetric variation caused by a user-defined variable for the Leksell Gamma Knife planning system. Treatment plans of 31 randomly selected patients were studied retrospectively to determine the dosimetric effects in the dose prescription and computation as a result of dose matrix positioning in the Leksell Gamma Plan (LGP, Version 4.12). Phantom studies with ion chamber measurements were carried out to validate the accuracy of the computation results. An average overdose of 2% was found due to the variations in the user defined dose matrix position for the studied cases. In the extreme, the overdose value was as high as 5% with an over-treatment time exceeding 2 min. The phantom measurements were found to agree with the LGP calculation within 0.5%. An adaptive method was developed and demonstrated in this study to eliminate such dosimetry variations. PMID- 10843115 TI - Intensity modulated photon beams subject to a minimal surface smoothing constraint. AB - A method to smooth intensity-modulated photon beams is presented which can be applied in conjunction with any optimization algorithm. The method employs an additional soft constraint to minimize the area of the surface defined by the photon fluence. PMID- 10843116 TI - Suppression of high-density artefacts in x-ray CT images using temporal digital subtraction with application to cryotherapy. AB - Image guidance in cryotherapy is usually performed using ultrasound. Although not currently in routine clinical use, x-ray CT imaging is an alternative means of guidance that can display the full 3D structure of the iceball, including frozen and unfrozen regions. However, the quality of x-ray CT images is compromised by the presence of high-density streak artefacts. To suppress these artefacts we applied temporal digital subtraction (TDS). This TDS method has the added advantage of improving the grey scale contrast between frozen and unfrozen tissue in the CT images. Two sets of CT images were taken of a phantom material, cryoprobes and a urethral warmer (UW) before and during the cryoprobe freeze cycle. The high density artefacts persisted in both image sets. TDS was performed on these two image sets using the corresponding mask image of unfrozen material and the same geometrical configuration of the cryoprobes and the UW. The resultant difference image had a significantly reduced artefact content. Thus TDS can be used to significantly suppress or eliminate high-density CT streak artefacts without reducing the metallic content of the cryoprobes. In vivo study needs to be conducted to establish the utility of this TDS procedure for CT assisted prostate or liver cryotherapy. Applying TDS in x-ray CT guided cryotherapy will facilitate estimation of the number and location of all frozen and unfrozen regions, potentially making cryotherapy safer and less operator dependent. PMID- 10843117 TI - High tibial osteotomy and ligament reconstruction for varus angulated anterior cruciate ligament-deficient knees. AB - In a consecutive series, we treated 41 young patients who had anterior cruciate ligament deficiency, lower limb varus angulation, and varying amounts of posterolateral ligament deficiency. Seventy-three percent of the patients (N = 30) had lost the medial meniscus and 63% (N = 26) had marked articular cartilage damage in the medial compartment. All patients were treated with high tibial osteotomy and, in the majority (N = 34), anterior cruciate ligament reconstruction a mean of 8 months later. Posterolateral reconstructions were also required in 18 knees. A 100% follow-up was obtained at a mean of 4.5 years after osteotomy. Gait analysis testing was done in 17 knees before and after osteotomy. At follow-up, a reduction in pain was found in 71% (29 knees); elimination of giving way, in 85% (35 knees); and resumption of light recreational activities without symptoms, in 66% (27 knees). The patient rating of the knee condition was normal or very good in 37% (15 knees) and good in 34% (14 knees). The mean Cincinnati Knee Rating Score significantly improved from 63 to 82 points. The mean adduction moment, 35% higher than controls preoperatively, significantly decreased to below normal values postoperatively. Correction of varus alignment was maintained in 33 knees (80%). We recommend osteotomy in addition to ligament reconstructive procedures in these knees with complex injury patterns. PMID- 10843118 TI - Intramuscular corticosteroid injection for hamstring injuries. A 13-year experience in the National Football League. AB - The purpose of this study was to assess the safety of intramuscular corticosteroid injection in selected, severe hamstring injuries in professional football players. Clinicians have been reluctant to use corticosteroid injections in or around muscle-tendon units because of concern of incomplete healing or rupture. We retrospectively reviewed the computer database of one National Football League team for all hamstring injuries requiring treatment between January 1985 and January 1998. We found that 431 players had suffered such injury. We developed a clinical grading system to identify hamstring injury severity and to stratify players for treatment. Fifty-eight players (13%) sustained severe, discrete injuries with a palpable defect within the substance of the muscle and were treated with intramuscular injection of corticosteroid and anesthetic. There were no complications related to the injection of corticosteroid. Only nine players (16%) missed any games as a result of their injury. Final examination revealed no strength deficits, normal muscle bulk and tone, and the ability to generate normal power. We believe that the grading system we developed can assist in selection of injury type for injection. Although lack of a control group limits statements of efficacy of injection, our impression is that intramuscular corticosteroid injection hastens players' return to full play and lessens the game and practice time they miss. PMID- 10843119 TI - Arthroscopically assisted mini-open rotator cuff repairs. Functional outcome at 2 to 7-year follow-up. AB - The functional outcome of 22 consecutive patients with full-thickness rotator cuff tears repaired using an arthroscopically assisted technique was evaluated. The average follow-up was 39 months (24 to 80), and the average tear size was 3.5 cm (1 to 7). There were 14 men and 8 women, with a mean age of 56 years (29 to 80); 86% of patients (N = 19) were satisfied with the results of surgery and 95% (N = 21) had improvement of their symptoms. All patients had a statistically significant improvement in pain and active abduction in the scapular plane and in external rotation. Postoperative strength in external rotation and abduction averaged 95% and 97% of the contralateral shoulder, respectively. Preoperative duration of symptoms, strength, age, and tear size were found to be independent predictors of outcome. The average Constant and Murley score was 84 of 100, the average American Shoulder and Elbow Surgeons score was 81 of 100, and the average University of California, Los Angeles, score was 31 of 35. Our results show that an arthroscopically assisted repair of full-thickness, moderate-to-large rotator cuff tears using uniform surgical technique and rehabilitation protocols provides excellent outcome with regard to function, pain, and activities of daily living. PMID- 10843120 TI - Portal-extension approach for the repair of small and medium rotator cuff tears. AB - One hundred ten consecutive cases (110 patients) of arthroscopically assisted rotator cuff repair through a limited, portal-extension approach were retrospectively reviewed. The average age of our patients was 58 years (range, 30 to 79). There were 35 women and 75 men. The dominant shoulder was affected in 67 patients (61%). All patients underwent a standard arthroscopic decompression. Acromioclavicular resections were performed in 15% of patients. The anterolateral portal was extended in the direction of Langer's lines to a total length of no more than 3 cm. The torn tendon was accessed through a small deltoid muscle split and repaired with nonabsorbable sutures. At an average follow-up of 35 months (range, 24 to 86), 106 patients (96%) had achieved excellent or satisfactory results. The average American Shoulder and Elbow Surgeons pain score improved from 7 preoperatively to 2 postoperatively. All but four patients were satisfied with the clinical result and reported significant improvement in active elevation and strength and a significant lessening of pain. Late acromioclavicular joint pain contributed to failure in three of the four patients with unsatisfactory results in this series. The results of this study suggest that, in selected patients with small to medium rotator cuff tears, arthroscopically assisted repair through an anterolateral portal-extension approach can produce excellent results. PMID- 10843121 TI - Lacrosse goalkeeper's thumb. A preventable injury. PMID- 10843122 TI - Injury profile in ice hockey from the 1970s through the 1990s in Finland. AB - The aim of this study was to investigate the incidence, types, and mechanisms of injury in Finnish ice hockey players at the highest competition level in different decades. Several teams were observed prospectively during the seasons between 1976 and 1979, and in the 1988 to 1989 and 1992 to 1993 seasons. An injury was defined as any sudden trauma requiring examination and treatment by a physician. The inclusion criteria were the same during the entire study. A total of 641 injuries were recorded. The injury rate per game increased significantly from 54 per 1000 player-hours in the 1970s to 83 per 1000 player-hours in the 1990s. The injury profile in the 1980s and 1990s differed from that in the 1970s. Per 1000 player-years, the rate of contusions as well as of sprains or strains increased significantly with each decade. Checking and unintentional collision with an opponent were common mechanisms of injury throughout the study, and the rate of injury by these mechanisms has continually increased. In conclusion, we suggest that there has been an increase in rough body contact between players, causing an alarming increase in the rate of ice hockey injuries. PMID- 10843123 TI - Comparison of traditional and subcutaneous patellar tendon harvest. A prospective study of donor site-related problems after anterior cruciate ligament reconstruction using different graft harvesting techniques. AB - Our goal was to compare the results after anterior cruciate ligament reconstruction using either the traditional one-incision or the subcutaneous two incision technique to harvest the central third of the patellar tendon, particularly concerning disturbances in anterior knee sensitivity and the patient's ability to walk on his or her knees. One surgeon performed anterior cruciate ligament reconstruction on 124 patients with unilateral ruptures and no history of previous incisions in the anterior knee region. The traditional one incision graft harvesting technique was used in 58 patients and the subcutaneous two-incision technique was used in 66 patients. At 2 years, the International Knee Documentation Committee classification, Lysholm score, arthrometry side-to side difference, and single-legged hop test showed no significant differences between groups. The area of insensitivity was a median of 24 cm2 in the traditional harvest group and 0 cm2 in the subcutaneous harvest group. The patients with subcutaneous harvest had a tendency toward fewer problems during walking on their knees than did the patients with traditional harvest. Our conclusion is that the subcutaneous two-incision graft harvesting technique caused less disturbance in anterior knee sensitivity and a tendency of less discomfort during walking on one's knees than the traditional one-incision technique. PMID- 10843124 TI - Longitudinal effects of anterior cruciate ligament injury and patellar tendon autograft reconstruction on neuromuscular performance. AB - We examined persons after anterior cruciate ligament injury and for 1.5 years after anterior cruciate ligament reconstruction to analyze changes in anterior knee laxity, lower extremity muscle strength, endurance, and several parameters of neuromuscular function. Sixteen men and nine women (average age, 23.8 years) were evaluated preoperatively, then underwent intraarticular autogenous patellar tendon anterior cruciate ligament reconstruction by the same surgeon and were evaluated at 6, 12, and 18 months postoperatively. Muscle strength was measured isokinetically and neuromuscular function was quantified with simultaneous anterior tibial translation and surface electromyography tests. Forty subjects (26 men and 14 women; average age, 23.5 years) with no known knee abnormalities served as the control group. Subjective questionnaire results showed that by 18 months postoperatively, 20 subjects (80%) believed they had regained their preoperative levels of function. Unfortunately, muscle function in most subjects had not returned to normal. At 12 to 18 months postoperatively, when knee rehabilitation was terminated, significant deficiencies in muscle performance persisted in most patients. Interestingly, in this group of stable knees, quadriceps and hamstring muscle reaction times appeared to be the best objective indicators of subjective knee function. PMID- 10843126 TI - Osteitis pubis and instability of the pubic symphysis. When nonoperative measures fail. AB - Seven rugby players with osteitis pubis and vertical instability at the pubic symphysis were treated operatively after nonoperative treatment had failed to improve their symptoms. The vertical instability was diagnosed based on flamingo view radiographs showing greater than 2 mm of vertical displacement. The players had undergone at least 13 months of nonoperative therapy before surgery was considered. Operative treatment consisted of arthrodesis of the pubic symphysis by bone grafting supplemented by a compression plate. At a mean follow-up of 52.4 months, all patients were free of symptoms and flamingo views confirmed successful arthrodesis with no residual instability of the pubic symphysis. Based on our results with this procedure, we believe that arthrodesis of the pubic symphysis has a role in the treatment of osteitis pubis that is recalcitrant to nonoperative treatment. The combination of osteitis pubis and vertical pubis symphyseal instability may be the cause of failure of nonoperative treatment. PMID- 10843125 TI - The effect of reconstruction of the medial patellofemoral ligament on patellar tracking. AB - We evaluated patellar tracking in six cadaveric knees with the medial restraints intact and then sectioned to determine their contribution to lateral translation of the patella with and without a lateral force on the patella. The medial patellofemoral ligament was then reconstructed with a gracilis tendon graft and patellar tracking was again evaluated. The knees were extended using a materials testing machine, and patellar tracking was measured with a position sensing system. With no lateral force applied to the patella, patellar tracking was unaffected by the presence or absence of the medial restraints or by reconstruction of the medial patellofemoral ligament. With a lateral force applied to the patella, patellar tracking was changed significantly by the loss of the medial restraints. Normal patellar tracking was substantially restored by reconstruction of the medial patellofemoral ligament. PMID- 10843127 TI - The influence of screw geometry on hamstring tendon interference fit fixation. AB - We used a standardized model of calf tibial bone to investigate the influence of screw diameter and length on interference fit fixation of a three-stranded semitendinosus tendon graft for anterior cruciate ligament reconstruction. Biodegradable poly-(L-lactide) interference screws with a diameter of 7, 8, and 9 mm and a length of 23 and 28 mm were used. We examined results in three groups of 10 specimens each: group 1, screw diameter equaled graft diameter and screw length was 23 mm; group 2, screw diameter equaled graft diameter plus 1 mm and screw length was 23 mm; group 3, screw diameter equaled graft diameter and screw length was 28 mm. The mean pull-out forces in groups 1, 2, and 3 were 367.2+/-78 N, 479.1+/-111.1 N, and 537.4+/-139.1 N, respectively. The force data from groups 2 and 3 were significantly higher than those from group 1. These results indicate that screw geometry has a significant influence on hamstring tendon interference fit fixation. Increasing screw length improves fixation strength more than oversizing the screw diameter. This is important, especially for increasing tibial fixation strength because the tibial graft fixation site has been considered to be the weak link of such a reconstruction. PMID- 10843128 TI - Baseball and softball sliding injuries. Incidence, and the effect of technique in collegiate baseball and softball players. AB - We prospectively observed seven softball and three baseball Division I collegiate teams to study the incidence of sliding injuries, the types of injuries resulting from the sliding technique, and the amount of time lost from participation. Slides were categorized as either feet- or head-first on the basis of the leading part of the body during the slide. Slides were further stratified depending on whether a diveback technique was performed. We recorded 37 injuries in 3889 slides in 637 games and 7596 athlete game exposures. The overall incidence of sliding injuries was 9.51 per 1000 slides and 4.87 per 1000 game exposures. Softball players had a significantly higher incidence of sliding injuries (12.13 per 1000 slides) than did baseball players (6.01 per 1000 slides). In baseball, the injury rate was higher for feet-first slides (7.31 per 1000 slides) than for headfirst slides (3.53 per 1000 slides) or divebacks (5.75 per 1000 divebacks). In softball, injury rates were higher for head-first slides (19.46 per 1000 slides) than for feet-first slides (10.04 per 1000 slides) or divebacks (7.49 per 1000 divebacks). The majority of injuries sustained were minor, with only four (11%) injuries causing the athlete to miss more than 7 days of participation. PMID- 10843129 TI - The effect of ciprofloxacin on tendon, paratenon, and capsular fibroblast metabolism. AB - The pathologic mechanisms underlying fluoroquinolone-induced tendinopathy are poorly understood. The observed incidence of tendinitis and tendon rupture in patients treated with ciprofloxacin hydrochloride suggests that the fluoroquinolone antibiotics alter tendon fibroblast metabolism. The purpose of this study was to examine the effect of ciprofloxacin on fibroblast metabolism in vitro. Canine Achilles tendon, paratenon, and shoulder capsule specimens were maintained in culture with ciprofloxacin (5, 10, or 50 microg/ml). Fibroblast proliferation, collagen synthesis, proteoglycan synthesis, and matrix-degrading activity were analyzed. Incubation of Achilles tendon, Achilles paratenon, and shoulder capsule fibroblasts with ciprofloxacin resulted in a statistically significant 66% to 68% decrease in cell proliferation compared with control cells at day 3 in culture. Ciprofloxacin caused a statistically significant 36% to 48% decrease in collagen synthesis compared with controls in all fibroblast cultures. Ciprofloxacin caused a statistically significant 14% to 60% decrease in proteoglycan synthesis in all fibroblast cell lines. Compared with unstimulated control fibroblasts, culture media from Achilles tendon, paratenon, and shoulder capsule cells that were exposed to ciprofloxacin demonstrated statistically significant increases in matrix-degrading proteolytic activity after 72 hours in culture. This study demonstrates that ciprofloxacin stimulates matrix-degrading protease activity from fibroblasts and that it exerts an inhibitory effect on fibroblast metabolism. The increase in protease activity and the inhibition of both cell proliferation and the synthesis of matrix ground substance may contribute to the clinically described tendinopathies associated with ciprofloxacin therapy. PMID- 10843130 TI - Contact mechanics of the medial tibial plateau after implantation of a medial meniscal allograft. A human cadaveric study. AB - The goal of this study was to determine how well a medial meniscal allograft restores the normal contact mechanics of the medial tibial plateau at the time of implantation. We measured maximum pressure, mean pressure, and contact area of the intact human cadaveric knee, the knee after meniscectomy, the knee with the original meniscus removed and reimplanted as an autograft, and the knee with an allograft. Measurements were made using pressure-sensitive film in 10 specimens loaded in compression to 1000 N at 0 degrees, 15 degrees, 30 degrees, and 45 degrees of flexion. The autograft and the allograft were identically implanted by cementing bone plugs attached to the meniscal horns in anatomic transtibial tunnels and suturing the outer edge of the meniscus to the remnant of the original meniscus. A medial meniscal allograft did not consistently restore normal contact mechanics because the process of implantation and the degree of match between the original and allograft meniscus affected the immediate load bearing performance of the transplant. However, the allograft did significantly reduce the contact pressure compared with the knee after meniscectomy. If the results from this study can be extrapolated to patients, then using an allograft to restore contact mechanics to normal may require improvements in surgical technique and graft selection. PMID- 10843131 TI - Outpatient open Bankart repair. AB - We evaluated satisfaction in 25 consecutive patients who underwent open Bankart repair as an outpatient procedure. The average age of the patients was 22 years. There were 18 men and 7 women. Patients were evaluated postoperatively via a home assessment questionnaire that included the following: home environment, dominant versus nondominant extremity, effectiveness of oral pain medication, ability to perform activities of daily living, and overall satisfaction. Three of the 25 patients would have preferred an overnight hospital stay. There were no admissions to the hospital or postoperative complications. The total charges for outpatient Bankart repair as compared with an inpatient procedure at the same institution resulted in a total cost savings of 56%. We conclude that outpatient Bankart repair is safe and economical. The overall cost savings were significant and patient satisfaction was 88%. PMID- 10843132 TI - Biomechanics of the coracoclavicular ligament complex and augmentations used in its repair and reconstruction. AB - Augmentation is a well-accepted and common component of coracoclavicular ligament repairs and reconstructions. The purpose of this study was to examine and compare the strength, stiffness, and mode of failure of the coracoclavicular ligament complex and four different augmentation techniques in cadaveric shoulders. There was no significant difference in the mean failure load between the intact ligament complex (724.9+/-230.9 N) and augmentations performed with braided polydioxanone (PDS) (676.7+/-115.4 N) or braided polyethylene placed through (986.1+/-391.1 N) or around (762.7+/-218.2 N) the clavicle. The mean failure load for augmentations using a 6.5-mm cancellous screw through the clavicle and into a single cortex of the coracoid (390.1+/-253.6 N) was significantly lower than that for the intact coracoclavicular ligaments. There was no difference in mean stiffness between the intact coracoclavicular ligament complex (115.9+/-36.2 N/mm) and the braided polyethylene augmentations placed through (99.8+/-22.2 N/mm) or around (90.0+/-25.5 N/mm) the clavicle. Polydioxanone augmentations were significantly less stiff (27.4+/-3.3 N/mm) than the intact complex, while screw augmentations were significantly stiffer (250.4+/-88.2 N/mm). There were no significant differences in strength or stiffness of braided polyethylene reconstructions placed around or through a drill hole in the clavicle. PMID- 10843133 TI - Sex-related injury patterns among selected high school sports. AB - This cohort observational study was undertaken to test the hypothesis that the incidence of injuries for girls participating in high school sports is greater than that for boys. From 1995 through 1997, players were included in our study if they were listed on the school's varsity team roster for boys' or girls' basketball, boys' or girls' soccer, boys' baseball, or girls' softball. Injuries and opportunities for injury were recorded daily. Certified athletic trainers reported injury and exposure data. Based on 39,032 player-seasons and 8988 reported injuries, the injury rates per 100 players for softball (16.7) and for girls' soccer (26.7) were higher than for baseball (13.2) and boys' soccer (23.4). The knee injury rates per 100 players for girls' basketball (4.5) and girls' soccer (5.2) were higher than for their male counterparts. Major injuries occurred more often in girls' basketball (12.4%) and soccer (12.1%) than in boys' basketball (9.9%) and soccer (10.4%). Baseball players (12.5%) had more major injuries than softball players (7.8%). There was a higher number of surgeries, particularly knee and anterior cruciate ligament surgeries, for female basketball and soccer players than for boys or girls in other sports. PMID- 10843134 TI - Patellar tendinopathy in athletes. Outcome of nonoperative and operative management. AB - We report the results of nonoperative and operative management of patellar tendinopathy in 42 athletes with Blazina stage 2 (26 patients) or stage 3 (16 patients) patellar tendinopathy. All patients were initially managed nonoperatively with nonsteroidal antiinflammatory drugs, physical therapy, and a progressive rehabilitation program based on isometric exercises, stretching, and eccentric exercises. After 6 months, 33 patients showed symptomatic improvement and were able to resume their sports. In nine patients with Blazina stage 3 tendinopathy, nonoperative measures failed, and surgery was performed. Operative treatment consisted of removal of the degenerated areas of the tendon, multiple longitudinal tenotomies, and drilling of the lower pole of the patella at the site of tendon attachment. Histologic examination of the excised tendon tissue showed areas of necrosis and mucoid degeneration, and alterations of the bone tendon junction. After a mean follow-up of 4.8 years, clinical results were excellent or good in all patients. In the group treated nonoperatively, results were better in the patients who had stage 2 tendinopathy than in those with stage 3. PMID- 10843135 TI - Periarticular heterotopic ossification after multiple knee ligament reconstructions. A report of three cases. AB - Heterotopic ossification is a frequently encountered clinical and radiographic entity. There are no previous reports in the English literature of heterotopic ossification after arthroscopically assisted ligament reconstructions for knee dislocations. Further, a link between the PCL reconstruction and posterior capsular ossification has not been heretofore recognized. Our three cases should raise the clinical awareness of such an entity. PMID- 10843136 TI - Partial rupture of the distal insertion of the patellar tendon. A report of two cases in professional athletes. PMID- 10843137 TI - Is the return to high-level athletics possible after fasciotomy for a compartment syndrome of the thigh? A case report. PMID- 10843138 TI - Symptomatic axillary artery dissection in a tennis player. Case report. PMID- 10843139 TI - Imaging techniques for the evaluation of glenohumeral instability. AB - The ability to image lesions associated with glenohumeral instability has evolved significantly over the past 2 decades. In the past, several imaging techniques ranging from conventional radiography to computerized axial arthrography and, most recently, to magnetic resonance imaging have been used to depict various labral abnormalities. In most instances, conventional radiography remains the initial imaging study for evaluating the patient with persistent shoulder pain and instability. Recently, however, magnetic resonance arthrography has been firmly established as the imaging modality of choice for demonstrating specific soft tissue abnormalities associated with glenohumeral instability. This article will review the role of various imaging modalities including conventional radiography, conventional arthrography, computerized axial arthrography, magnetic resonance imaging, and magnetic resonance arthrography. Emphasis will be placed on the role of magnetic resonance arthrography as it pertains to the lesions associated with glenohumeral instability. A thorough discussion of the appearance of normal anatomic structures, anatomic variations that mimic abnormality, and the various lesions associated with glenohumeral instability will be provided. PMID- 10843140 TI - Rib stress fractures in elite rowers. A case series and proposed mechanism. PMID- 10843141 TI - Rigorous statistical reliability, validity, and responsiveness testing of the Cincinnati Knee Rating System in 350 subjects with uninjured, injured, or anterior cruciate ligament-reconstructed knee. PMID- 10843142 TI - Arthroscopic reconstruction of the anterior cruciate ligament. A comparison of patellar tendon autograft and four-strand hamstring tendon autograft. PMID- 10843143 TI - Team physician consensus statement. PMID- 10843144 TI - Cell adhesion molecules in cardiovascular disease and its risk factors--what can soluble levels tell us? PMID- 10843145 TI - Effects of hyperinsulinemia on plasma levels of circulating adhesion molecules. AB - Plasma levels of circulating intercellular adhesion molecule-1 (cICAM-1), a potential cardiovascular risk factor, are increased in diabetics. Among other factors, hyperinsulinemia has been proposed to enhance its release into the circulation. Thus, we directly examined the effects of insulin infusion on plasma levels of circulating adhesion molecules, and two other endothelial markers, i.e. von Willebrand factor (vWF) and soluble thrombomodulin (sTM). The study design was balanced, randomized, placebo-controlled, double blind, and cross-over. Twelve healthy male subjects received, on separate study days, a euglycemic hyperinsulinemic clamp (3 mU/kg x min) or placebo over 6 h. Plasma levels of cICAM-1, vascular cell adhesion molecule-1, circulating E-selectin, and sTM were measured by enzyme immunoassay; vWF-Ag was measured using a STA clot analyzer. Plasma levels of these adhesion molecules and endothelial cell activation markers were not affected despite a 30-fold increase in insulin levels. Hyperinsulinemia has no adverse effect on circulating ICAM-1, vascular cell adhesion molecule-1, E selectin, vWF, or sTM and therefore does not directly induce endothelial activation. PMID- 10843146 TI - Dissecting the genetics of hyperparathyroidism--new clues from an old friend. PMID- 10843147 TI - Familial papillary thyroid cancer--many syndromes, too many genes? PMID- 10843148 TI - Papillary thyroid carcinoma associated with papillary renal neoplasia: genetic linkage analysis of a distinct heritable tumor syndrome. AB - Papillary thyroid carcinoma usually is sporadic, but may occur in a familial form. The complete clinical and pathological phenotype of familial papillary thyroid carcinoma (fPTC) has not been determined, and the susceptibility gene(s) is unknown. We investigated the clinical and pathological characteristics of an unusually large three-generation fPTC kindred to characterize more fully the clinical phenotype. We performed linkage analysis to determine the chromosomal location of a fPTC susceptibility gene. In addition to the known association of fPTC with nodular thyroid disease, we observed the otherwise rare entity of papillary renal neoplasia (PRN) in two kindred members, one affected with PTC and the other an obligate carrier. The multifocality of PRN in one subject adds weight to the likelihood of a true genetic predisposition to PRN. Both genetic linkage and sequence analysis excluded MET, the protooncogene of isolated familial PRN, as the cause of the fPTC/PRN phenotype. A genome-wide screening and an investigation of specific candidate genes demonstrated that the fPTC/PRN phenotype was linked to 1q21. A maximum three-point log of likelihood ratio score of 3.58 was observed for markers D1S2343 and D1S2345 and for markers D1S2343 and D1S305. Critical recombination events limited the region of linkage to approximately 20 cM. A distinct inherited tumor syndrome has been characterized as the familial association of papillary thyroid cancer, nodular thyroid disease, and papillary renal neoplasia. The predisposing gene in a large kindred with this syndrome has been mapped to 1q21. PMID- 10843149 TI - Fulminant hepatitis A in a patient with severe hyperthyroidism: rapid recovery from hepatic coma after plasmapheresis and total thyroidectomy. PMID- 10843150 TI - Leptin in postmenopausal women: influence of hormone therapy, insulin, and fat distribution. AB - Whether use of hormone-replacement therapy (HRT) influences menopause-related changes in body weight is unclear. HRT may affect energy balance by influencing synthesis of the adipocyte-derived hormone leptin. The objectives of this study were to: 1) identify factors influencing circulating leptin in postmenopausal women; 2) determine whether HRT influences serum leptin after adjusting for confounding factors; and, 3) identify potential independent effects of HRT or leptin on resting energy expenditure (REE). Subjects were 54 postmenopausal women, 45-55 yr old, 35 of whom used HRT (estrogen plus progestin). Total and regional body composition and fat distribution were determined by dual-energy x ray absorptiometry and computed tomography; fasting serum leptin and insulin, by RIA; and REE, by indirect calorimetry. Stepwise multiple linear regression analysis indicated that serum leptin could best be predicted from total fat mass, fasting serum insulin, and total lean mass [log leptin = 1.08 x log fat mass) + (0.46 x log insulin) + (-1.25 x log lean mass) + 1.88; model R2 = 0.78, P < 0.001]. Multiple linear regression analysis indicated that visceral fat was independently related to leptin (parameter estimate = 0.23, P < 0.05), after adjusting for s.c. abdominal fat and leg fat, as well as lean mass and insulin. After adjusting for total fat mass, total lean mass, and fasting insulin, serum leptin did not differ between users and nonusers of HRT (21.7 +/- 1.0 vs. 20.2 +/ 1.3 ng/mL, P = 0.369, adjusted mean +/- SE, respectively). Serum estradiol was inversely correlated with (adjusted) leptin in non-HRT users (r = -0.50), suggesting that ovarian senescence may lead to an increase in leptin. Multiple linear regression analysis indicated that REE (adjusted for fat mass, fat-free mass, and ethnicity) was not associated with leptin (P = 0.298) or hormone use status (P = 0.999; 1323 +/- 31 vs. 1316 +/- 42 kcal/day, adjusted mean +/- SE for users and nonusers, respectively). These results indicate that, in postmenopausal women: 1) total fat mass, lean mass, and fasting insulin, but not HRT, are significant determinants of serum leptin; 2) visceral and s.c. fat contribute to serum leptin; and, 3) neither HRT nor leptin is independently related to REE. PMID- 10843151 TI - Gender differences in the prevalence of metabolic complications in familial partial lipodystrophy (Dunnigan variety). AB - Familial partial lipodystrophy, Dunnigan type (FPLD; Mendelian Inheritance in Man #151660), is an autosomal dominant disorder characterized by loss of s.c. fat from the extremities and trunk since puberty and predisposition to insulin resistance and its complications. However, for lack of recognition of affected men, previous studies could not ascertain any gender differences in phenotypic expression. Therefore, anthropometric variables and prevalence of diabetes mellitus, dyslipidemia, hypertension, and atherosclerotic vascular disease were compared among 17 postpubertal men and 22 women with FPLD from eight pedigrees. All individuals completed a questionnaire, and fasting blood was analyzed for glucose, insulin, and lipoprotein concentrations. Both affected men and women had similar patterns of fat loss. Compared with the affected men, women had higher prevalence of diabetes (18% and 50%, respectively; P = 0.05) and atherosclerotic vascular disease (12% and 45%, respectively; P = 0.04) and had higher serum triglycerides (median values, 2.27 and 4.25 mmol/L, respectively; P = 0.02) and lower high-density lipoprotein cholesterol concentrations (age-adjusted means, 0.94 and 0.70 mmol/L, respectively; P = 0.04). The prevalence of hypertension and fasting serum insulin concentrations were similar. In conclusion, women with FPLD are more severely affected with metabolic complications of insulin resistance than men. These observations raise the possibility that women with generalized and regional obesity may also have more severe metabolic sequelae of insulin resistance. PMID- 10843152 TI - Comparison of alendronate and intranasal calcitonin for treatment of osteoporosis in postmenopausal women. AB - This study compared the effects of oral alendronate and intranasal calcitonin for treatment of osteoporosis in postmenopausal women. Women at least 5 yr postmenopause (n = 299) were randomized to either 10 mg alendronate, matching alendronate placebo, or open-label intranasal calcitonin 200 IU daily for 12 months. Hip and spine bone mineral density (BMD) and markers of bone turnover were measured, and safety and tolerability were assessed. Alendronate produced greater increases in BMD than calcitonin at 12 months at the lumbar spine (5.16% vs. 1.18%; P < 0.001), trochanter (4.73% vs. 0.47%; P < 0.001), and femoral neck (2.78% vs. 0.58%; P < 0.001). Changes in BMD with calcitonin were greater than with placebo at the femoral neck, but were not different from placebo at either the trochanter or lumbar spine. Greater decreases in bone turnover were seen with alendronate than with calcitonin (serum bone-specific alkaline phosphatase, 43% vs. 9%, P < 0.001; urinary N-telopeptide, 62% vs. 11%, P < 0.001). Similar percentages of patients in each group reported an adverse experience during the study. We conclude that, in postmenopausal women with osteoporosis, 12 months of therapy with alendronate produced significantly greater increases in BMD of the hip and spine and greater decreases in bone turnover than intranasal calcitonin. PMID- 10843153 TI - The dominant role of increased intrasellar pressure in the pathogenesis of hypopituitarism, hyperprolactinemia, and headaches in patients with pituitary adenomas. AB - Mild hyperprolactinemia frequently accompanies the hypopituitarism seen in patients with pituitary macroadenomas that do not secrete PRL. Recent data suggested that the hypopituitarism and mild hyperprolactinemia in this setting are largely due to compression of pituitary stalk and portal vessels. Headaches (HAs) are frequently seen in patients with large adenomas and at times in those with microadenomas. Because the walls of the sella turcica are relatively rigid, we postulate that tumor growth within the sella increases intrasellar pressure (ISP), which in turn impairs portal blood flow, resulting in mild hyperprolactinemia and hypopituitarism. We also postulate that increased mean ISP (MISP) contributes to the development of HAs. Normal MISP is not known but is unlikely to exceed normal intracranial pressure of less than 10-15 mm Hg. We determined MISP in 49 patients who had transsphenoidal surgery for pituitary adenomas. MISP was measured using a commonly available intracranial monitoring kit where a fiberoptic transducer was inserted through a 2-mm dural incision at the time of adenomectomy. Patients with deficient FSH, LH, ACTH, or TSH secretion were considered hypopituitary. Data on serum PRL levels were included for analysis only in patients whose adenomas had negative immunostaining for the hormone. MISP measurements ranged from 7-56 mm Hg, with a mean (+/-SD) of 28.8 +/ 13.5 and a median of 26 mm Hg. The pressure measurements were higher in patients with hypopituitarism than in those with normal pituitary function (P = 4.6013 x 10(-6)). Patients presenting with HAs had higher MISP than those who did not (P = 5.44 x 10(-7)), regardless of their pituitary function or tumor sizes. PRL levels correlated positively with MISP values (r = 0.715, P < 0.0001). Tumor size did not correlate with MISP or PRL levels. The findings of increased MISP in hypopituitary patients and the documented correlation with PRL levels, suggest that ISP is a major mechanism involved in the pathogenesis of hypopituitarism and hyperprolactinemia. Similarly, the increased MISP in patients with HAs, irrespective of tumor size or pituitary function, suggest that increased ISP is a major mechanism involved in the pathogenesis of this symptom. The data support the hypothesis that in patients with pituitary adenomas increased ISP is a major mechanism contributing to the development of hyperprolactinemia, hypopituitarism, and HAs. Increased ISP in these patients leads to compression of the portal vessels and the associated interruption of the delivery of hypothalamic hormones to the anterior pituitary. This would explain the reversibility of pituitary function observed in most patients after adenomectomy. However, increased ISP may also lead to decreased blood supply, resulting in ischemic necrosis in some regions of the pituitary. The latter could limit potential recovery of pituitary function after adenomectomy. PMID- 10843154 TI - Decrease in gonadotropin-releasing hormone (GnRH) pulse frequency with aging in postmenopausal women. AB - Increasing evidence suggests that aging is associated with dynamic changes in the hypothalamic and pituitary components of the reproductive axis that are independent of changes in gonadal hormone secretion. This study was designed to determine the effect of age on GnRH pulse frequency in women in the absence of gonadal feedback using gonadotropin free alpha-subunit (FAS) and LH as neuroendocrine markers of endogenous GnRH secretion. All studies were performed in healthy, euthyroid postmenopausal women (PMW) during daytime hours. The impact of sampling interval and duration on assessment of pulse frequency in PMW was first examined in 10 women with a mean age of 61.6 +/- 8 yr (mean +/- SD), in whom blood was sampled every 5 min for 12 h. Each 5-min series was then reduced to simulate a 10-min series and then a 15-min series for pulse analysis, and the effect of 8 h compared with 12 h of sampling was determined. To define the changes in the frequency and amplitude of pulsatile hormone secretion with aging, 11 younger (45-55 yr) and 11 older (70-80 yr) PMW were then studied over 8 h at a 5-min sampling interval. In the initial series, the mean interpulse intervals (IPIs) for FAS were 53.8 +/- 3.6, 69.2 +/- 3.9, and 87.6 +/- 7.3 min at sampling intervals of 5, 10, and 15 min, respectively (P < 0.0005). The LH IPI also increased progressively with sampling intervals of 5, 10, and 15 min (54.4 +/- 2.5, 70.4 +/- 2.3, and 91.1 +/- 4.4 min; P < 0.0001). At the 5-min sampling interval, the calculated number of pulses/24 h was not different between a 12-h series compared with an 8-h series for either FAS or LH. In the second series of studies, the older PMW had lower gonadotropin levels (LH, 86.5 +/- 8.8 vs. 51.3 +/- 7.7 IU/L, P < 0.01; FSH, 171.6 +/- 16.9 vs. 108.2 +/- 10.5 IU/L, P < 0.005; FAS, 1021.5 +/- 147.4 vs. 425.6 +/- 89.6 ng/L, P < 0.005, in younger and older PMW, respectively) despite no differences in estrone or estradiol levels. The older PMW also demonstrated a slower FAS pulse frequency compared with their younger counterparts, as reflected in an increased FAS IPI (52.6 +/- 3.1 and 70.6 +/- 5.9 min; P < 0.002). The difference in IPIs between younger and older PMW was not statistically significant for LH (65.4 +/- 5.6 and 71.8 +/- 6.6 min for younger and older PMW, respectively). FAS pulse amplitude was decreased in older PMW compared with younger PMW (431.7 +/- 66.2 vs. 224.6 +/- 81.9 ng/L; P < 0.01), whereas the decrease in LH pulse amplitude with age was of borderline statistical significance (23.2 +/- 3.1 vs. 15.9 +/- 2.1 IU/L; P = 0.09). IN CONCLUSION: 1) the use of a 5-min sampling interval and measurement of FAS as the primary marker of GnRH pulse generator activity indicate that GnRH pulse frequency in younger PMW is faster than previously reported, but not increased over that seen in the late follicular phase and midcycle surge in women with intact ovarian function; and 2) the marked decrease in FAS pulse frequency with age provides evidence of age-related changes in the hypothalamic component of the reproductive axis that are independent of changes in gonadal function. PMID- 10843155 TI - Significance of Pro12Ala mutation in peroxisome proliferator-activated receptor gamma2 in Korean diabetic and obese subjects. AB - Peroxisome proliferator-activated receptors (PPARs) are a nuclear hormone receptor superfamily of ligand-activated transcription factors, and the PPARgamma subtype regulates adipocyte differentiation, lipid metabolism, and insulin sensitivity. There have been several reports on the relationship between the PPARgamma2 Pro12Ala genotype and obesity or diabetes in Caucasians. The objective of this study was to examine the relationship between this mutation and obesity or diabetes in Korean subjects. Two hundred and twenty-nine Korean subjects, including 111 obese subjects (body mass index, >25 kg/m2) were included in this study. One hundred and eleven subjects had normal glucose tolerance, 60 had impaired glucose tolerance, and 58 had diabetes mellitus. We evaluated these subjects for the Pro12Ala mutation in the PPARgamma gene using PCR-restriction fragment length polymorphism. Allele frequencies of the Pro12Ala missense mutation of PPARgamma2 were not different among Korean subjects with normal glucose tolerance (qAla = 0.045), those with impaired glucose tolerance (qAla = 0.033), and those with diabetes mellitus (qAla = 0.043; P > 0.05). Allele frequencies of PPARgamma2 Ala in obese subjects (qAla = 0.036) were not significantly different from those in nonobese subjects (qAla = 0.047). These results suggest that the Pro12Ala mutation in PPARgamma is not associated with either diabetes or obesity and may not be an important determinant of obesity or diabetes in Korean subjects. PMID- 10843156 TI - Calcium absorption, bone mass accumulation, and kinetics increase during early pubertal development in girls. AB - To evaluate the changes in calcium and bone mineral metabolism associated with early pubertal development, we performed longitudinal measurements of calcium absorption, calcium kinetics, bone mineral content, and hormonal markers related to puberty in a multiethnic group of girls beginning when they were 7 or 8 yr old. Girls were Tanner stage 1 (breast) at the start of the study. They were placed on a 1200 mg/day dietary calcium intake and studied at approximately 6 month intervals until they reached Tanner stage 2 (breast). Results at that time point (PUB) were compared to values obtained approximately 1 yr earlier (LatePRE) and those 1 yr before that (EarlyPRE). We found an increase in calcium absorption comparing PUB to LatePRE (n = 34; 36.6 +/- 8.7% vs. 30.7 +/- 9.9%; P = 0.002). Using whole body, dual energy, x-ray absorptiometry scanning, we found an increase in calcium gain during the LatePRE to PUB period compared with that during the EarlyPRE to LatePRE period (135 +/- 53 vs. 110 +/- 45 mg/day; P = 0.04). Calcium kinetic studies showed a significant increase in the bone calcium deposition rate (Vo+) during the PUB compared to the LatePRE period. Hormonal and biochemical markers of bone development were also significantly increased at PUB compared to LatePRE. Hormonal activity, as evidenced by the unstimulated LH level, was significantly correlated with calcium gain between the LatePRE and PUB studies and the bone calcium deposition rate in the PUB study. These data demonstrate, using multiple independent methods, an increase in calcium utilization associated with the earliest physical signs of puberty. PMID- 10843157 TI - Plasma leptin concentrations and cardiac autonomic nervous system in healthy subjects with different body weights. AB - Previous studies have shown that leptin stimulates sympathetic nervous system; heart rate variability (HRV) is a widely used technique for assessing the sympathovagal balance at the cardiac level. The aim of our study was to investigate a possible relationship between plasma leptin levels and the autonomic regulation using spectral analysis of HRV. In 120 healthy nonobese subjects the plasma leptin concentration was determined, and HRV was recorded at baseline and during tilt. All subjects were categorized in quartiles of plasma leptin concentration. Analysis of data showed a significant increase in body mass index, body fat, fasting plasma insulin, triglyceride concentration, and homeostatic model assessment values throughout the different quartiles of plasma leptin concentration. Concerning cardiovascular parameters, heart rate, arterial blood pressures, and RR intervals were not significantly different among the quartiles. Total power and high frequency (HF) in normalized units were significantly decreased, whereas low frequency (LF) normalized units was progressively increased from the first to the fourth quartile. Thus, the LF/HF ratio rose gradually and significantly from the lowest to the highest quartile. Such results were independent of the body fat estimate (P < 0.03 for the trend). The change in the LF/HF ratio was significantly enhanced during tilt (P < 0.001 vs. rest values for all quartiles); the effect was stronger in subjects in the fourth quartile of plasma leptin concentration (P < 0.005 for the trend). The latter parameter was also independent of body fat content and distribution (P < 0.01). Our study shows that increasing fasting plasma leptin concentrations are associated with a shift of the sympathovagal balance toward a progressive increase in sympathetic activation and an increased response to orthostatic stimulus in nonobese subjects with different body fat contents. PMID- 10843158 TI - Cardiovascular effects of parathyroid hormone: a study in healthy subjects and normotensive patients with mild primary hyperparathyroidism. AB - The aim of the study was to evaluate: 1) the cardiovascular function and the autonomic drive to the heart in patients affected by primary hyperparathyroidism (pHPT) with no evidence of renal and cardiovascular complications; 2) the cardiovascular effects of acute administration of PTH in normal subjects. In 14 patients affected by mild asymptomatic pHPT echocardiographic assessment of cardiovascular function and of the mechanic properties of the brachial and carotid artery, heart rate variability and the dispersion of QT interval were performed before and 6 months after successful surgery. Twenty age- and sex matched healthy subjects were included in the study. Five healthy volunteers underwent a single blind, placebo-controlled, random order, cross-over study with infusion of PTH (hPTH 1-34, 200 U in saline over 5 min) or placebo. Echocardiographic assessment of cardiovascular function, heart rate variability, and QT interval were performed between 20 and 25 min after the start of the infusion and repeated after 15 min of tilting at 60 degrees. In pHPT patients the echocardiographic parameters were normal; left ventricular isovolumetric relaxation time was always in the normal range, but significantly shorter than in control subjects, suggesting an increased sympathetic stimulation. Arterial diameters and thickness, blood pressure, and QT interval were not significantly different with respect to normal subjects and were unchanged 6 months after surgery. pHPT patients lacked the circadian rhythm of the low frequency to high frequency ratio, suggesting an increased sympathetic drive to the heart at nighttime. In normal subjects there were no significant differences in basal echocardiographic measurements during PTH infusion with respect to placebo and in the hemodynamic response to tilt. These results suggest that cardiovascular function is substantially normal in normotensive pHPT patients with mild hypercalcemia. A modulation of the adrenergic control of circulation seems to be associated with hypercalcemia and/or chronic PTH excess, but its biological relevance needs further investigations. PMID- 10843159 TI - Effect of thyroid hormones on cardiac function, geometry, and oxidative metabolism assessed noninvasively by positron emission tomography and magnetic resonance imaging. AB - Thyroid hormones influence cardiac performance directly and indirectly via changes in peripheral circulation. Little, however, is known about the effect on myocardial oxidative metabolism and its relation to cardiac function and geometry. Patients with a history of thyroidectomy for thyroid cancer present a unique model to investigate the cardiac effects of hypothyroidism. Ten patients without heart disease were investigated in the hypothyroid state and again 4-6 weeks later under euthyroid conditions. Myocardial oxidative metabolism was measured by positron emission tomography with [11C]acetate and the clearance constant k(mono). Cine magnetic resonance imaging was applied to determine left ventricular geometry. A stroke work index (SWI = stroke volume x systolic blood pressure/ventricular mass) was calculated. Then, to estimate myocardial efficiency, a work metabolic index [WMI = SWI x heart rate/k(mono)] was obtained. Compared to hormone replacement, systemic vascular resistance and left ventricular mass were significantly higher in hypothyroidism. Ejection fraction and SWI were significantly lower. Despite an additional reduction of k(mono), the WMI was significantly lower, too. In summary, cardiac oxygen consumption is reduced in hypothyroidism. This reduction is associated with increased peripheral resistance and reduced contractility. Estimates of cardiac work are more severely suppressed than those of oxidative metabolism, suggesting decreased efficiency. These findings may provide an explanation for development or worsening of heart failure in hypothyroid patients with preexisting heart disease. PMID- 10843160 TI - Preeclampsia is associated with low circulating levels of insulin-like growth factor I and 1,25-dihydroxyvitamin D in maternal and umbilical cord compartments. AB - Insulin-like growth factor I (IGF-I) stimulates renal and placental 1,25 dihydroxyvitamin D [1,25-(OH)2D] and is considered an important regulator of fetal growth. As 1,25-(OH)2D and birth weight are low in preeclampsia, this study was undertaken to determine whether circulating levels of IGF-I were associated with serum 1,25-(OH)2D concentrations in preeclamptic (PE group) and normotensive (NT group) pregnancies. Maternal and umbilical cord serum levels of IGF-I and 1,25-(OH)2D were significantly (P < 0.01) lower in the PE group than in the NT group. The concentrations of these two hormones correlated significantly in the umbilical cord (P < 0.05) and in the maternal (P < 0.001) compartments of the PE and NT groups, respectively. The amount of IGFBP-3 was 64% lower whereas that of IGFBP-1 was 2.9-fold higher in umbilical cord serum of the PE group compared with the NT group. In addition, maternal and umbilical cord serum IGF-I correlated significantly (P < 0.05) with weight and length at birth only in the PE group. In conclusion, the results of this study indicate that circulating IGF-I and 1,25 (OH)2D levels in both maternal and umbilical cord compartments are low in preeclampsia. Furthermore, this study suggests a differential regulatory effect of IGF-I on 1,25-(OH)2D synthesis and fetal growth depending on the presence or absence of preeclampsia. PMID- 10843161 TI - The plasma levels of dehydroepiandrosterone sulfate are decreased in patients with chronic heart failure in proportion to the severity. AB - Dehydroepiandrosterone sulfate (DHEAS) is the major secretory steroid of the human adrenal glands. The secretion of DHEAS decreases with aging. The incidence of heart failure also rises in the elderly population. We measured the plasma levels of DHEAS and cortisol in 49 patients with chronic heart failure (CHF) and 32 age-matched controls and assessed its relation to plasma levels of A-type natriuretic peptide and B-type natriuretic peptide, biochemical markers of heart failure. Plasma levels of DHEAS were significantly lower in patients with CHF than in controls, whereas there was no significant difference in plasma levels of cortisol between the two groups. In stepwise regression analysis, the plasma level of DHEAS was significantly and independently correlated with age (beta = 0.451; P < 0.0001) and the plasma level of B-type natriuretic peptide (beta = 0.338; P < 0.001), and the plasma cortisol/DHEAS ratio was significantly and independently correlated with the plasma levels of A-type natriuretic peptide (beta = 0.598; P < 0.0001) and thiobarbituric acid-reactive substances (a marker of oxidative stress; beta = 0.252; P < 0.01) and age (beta = 0.171; P < 0.05). These results indicate that the plasma levels of DHEAS are decreased in patients with CHF in proportion to its severity and that oxidative stress is associated with decreased levels of DHEAS in patients with CHF. PMID- 10843162 TI - Estrogen replacement therapy in a man with congenital aromatase deficiency: effects of different doses of transdermal estradiol on bone mineral density and hormonal parameters. AB - The effects of different doses of transdermal estradiol (TE) on bone mineral density (BMD) in a man with aromatase deficiency were evaluated. The study protocol was divided in the following four phases: phase 1, before estradiol treatment; phase 2, 50 microg TE twice weekly for 6 months; phase 3, 25 microg TE twice weekly for 9 months; and phase 4, 12.5 microg TE twice weekly for 9 months. X-rays of hands, legs, and pelvis were performed, and BMD of the lumbar spine, hormonal parameters (LH, FSH, testosterone, and estradiol), and markers of bone turnover were determined during each phase. BMD in phase 1 was 0.933 g/cm2 and increased to 1.051 and 1.173 g/cm2 after 4 and 7 months of TE, respectively. In phase 3, BMD reached the maximum value (1.275 g/cm2). In phase 4, BMD decreased to 1.180 g/cm2 and was 1.029 g/cm2 at the end of the study protocol. A bilateral necrosis of femoral heads was also detected by x-ray films. In phase 1 serum testosterone was in the normal range, whereas serum estradiol was undetectable. During the 24-month period of treatment with TE (phases 2-4), estradiol was directly related to the amount of TE, whereas LH was inversely related to estradiol serum levels. Estradiol and gonadotropins reached optimal values only in phase 3, when FSH also was near normal; serum testosterone concentrations were normal in phases 3 and 4. This study confirms the role of estrogens in achieving and maintaining bone mineral content in the human male, providing further clinical tools useful in the management of bone loss in aromatase deficiency in the male. We suggest that the adequate substitutive dose of TE for maintaining both bone mass and normal estradiol serum levels in adult men with aromatase deficiency may be 25 microg twice weekly (0.47 microg/kg weekly). PMID- 10843163 TI - Pamidronate treatment of severe osteogenesis imperfecta in children under 3 years of age. AB - Severe osteogenesis imperfecta (OI) is a hereditary disorder characterized by increased bone fragility and progressive bone deformity. Cyclical pamidronate infusions improve clinical outcome in children older than 3 yr of age with severe OI. Because earlier treatment may have potential to prevent deformities and improve functional prognosis in young children, we studied nine severely affected OI patients under 2 yr of age (2.3-20.7 months at entry) for a period of 12 months. Pamidronate was administered i.v. in cycles of 3 consecutive days. Patients received four to eight cycles during the treatment period, with cumulative doses averaging 12.4 mg/kg. Clinical changes were evaluated regularly during treatment, and radiological changes were assessed after 6-12 months of treatment. The control group consisted of six age-matched, severely affected OI patients, who had not received pamidronate treatment. During treatment bone mineral density (BMD) increased between 86-227%. The deviation from normal, as indicated by the z-score, diminished from -6.5 +/- 2.1 to -3.0 +/- 2.1 (P < 0.001). In the control group the BMD z-score worsened significantly. Vertebral coronal area increased in all treated patients (11.4 +/- 3.4 to 14.9 +/- 1.8 cm2; P < 0.001), but decreased in the untreated group (P < 0.05). In the treated patients, fracture rate was lower than in control patients (2.6 +/- 2.5 vs. 6.3 +/- 1.6 fractures/year; P < 0.01). No adverse side-effects were noted, apart from the well known acute phase reaction during the first infusion cycle. Pamidronate treatment in severely affected OI patients under 3 yr of age is safe, increases BMD, and decreases fracture rate. PMID- 10843164 TI - Normal endothelial function despite insulin resistance in healthy women with the polycystic ovary syndrome. AB - Women with the polycystic ovarian syndrome (PCOS) carry a number of cardiovascular risk factors, including insulin resistance, lipid abnormalities, and an altered pattern of sex steroid exposure. Noninvasive measurements of endothelial function, which can demonstrate abnormalities well in advance of clinically apparent disease, have not been previously reported in this patient group. We undertook a cross-sectional evaluation of endothelium-dependent and independent vascular function using brachial artery ultrasound. We studied healthy women with clinical and laboratory evidence of PCOS (n = 18) and age matched controls (n = 19), not taking any antihypertensive, cholesterol-lowering, or hormonal therapies. Laboratory parameters of insulin resistance, glycemia, cholesterol status, and hormone levels were also measured. Despite marked differences in glucose/insulin ratio [6.1 +/- 1.1 mmol/pmol (PCOS) vs. 9.9 +/- 0.6 (controls)] and free androgen index [11.9 +/- 2.3 (PCOS) vs. 3.7 +/- 0.6 (controls); normal, <5], we did not find evidence of impaired endothelial function in our patients with PCOS. Both endothelium-dependent (8.7 +/- 3.1%) and endothelium-independent (23.2 +/- 3.4%) vascular responses were normal, and practically identical to the responses seen in the control group (endothelium dependent, 9.0 +/- 0.7; endothelium-independent, 23.0 +/- 1.2%). The PCOS women were more obese, but baseline brachial arterial diameters were not different between groups. There was no correlation between degree of insulin resistance or hyperandrogenism and the brachial response. This group of healthy obese young women with insulin resistance and hyperandrogenism due to PCOS had normal endothelium-dependent and -independent vascular responses compared to age-matched controls. The factors resulting in preservation of these response are unclear and warrant further investigation. PMID- 10843165 TI - Changes in plasma low-density lipoprotein (LDL)- and high-density lipoprotein cholesterol in hypo- and hyperthyroid patients are related to changes in free thyroxine, not to polymorphisms in LDL receptor or cholesterol ester transfer protein genes. AB - Thyroid function disorders lead to changes in lipoprotein metabolism. Both plasma low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) increase in hypothyroidism and decrease in hyperthyroidism. Changes in LDL-C relate to altered clearance of LDL particles caused by changes in expression of LDL receptors on liver cell surfaces. Changes in cholesterol ester transfer activity partly explain changes in HDL-C. It has been suggested that the magnitude of these changes is related to polymorphisms of involved genes. The aim of the present study is to investigate whether the polymorphic AvaII restriction site in exon 13 of the LDL receptor gene and the polymorphic TaqIB site in intron 1 of the cholesterol ester transfer protein are associated with the magnitude of the changes in plasma LDL-C and HDL-C, respectively, in the transition from the hypo- or hyperthyroid to the euthyroid state. From a consecutive group of 66 untreated hypothyroid and 60 hyperthyroid patients, 47 Caucasians in each group were analyzed. Fasting LDL-C and HDL-C were measured at baseline and 3 months after restoration of the euthyroid state. Genotype was determined by means of PCR techniques. The homozygous presence of a restriction site was designated as +/+, heterozygous as +/-, and absence as -/-. Trend analysis was done with ANOVA. Among hypo- or hyperthyroid patients, subgroups with different genotypes did not differ in thyroid function pre- or post treatment. The mean decrease in LDL-C (mmol/L +/- SD) in hypothyroid patients with different AvaII genotypes did not differ: - 1.07 +/- 1.44 (-/-, N = 15), 1.25 +/- 1.53 (+/-, N = 19), and -1.18 +/- 1.01 (+/+, N = 13) mmol/L [not significant (NS)]; neither did the mean increase in hyperthyroid patients: 1.07 +/- 0.90 (-/-, N = 18), 0.92 +/- 1.00 (+/-, N = 21), and 1.20 +/- 0.45 (+/+, N = 6) (NS). The mean decrease in HDL-C (mmol/L +/- SD) in hypothyroid patients with different TaqIB genotypes did not differ: -0.22 +/- 0.26 (-/-, N = 13), -0.15 +/- 0.23 (+/-, N = 21), and -0.12 +/- 0.22 (+/+, N = 9) (NS); neither did the mean increase in hyperthyroid patients: 0.29 +/- 0.39 (-/-, N = 7), 0.26 +/- 0.23 (+/ , N = 22), and 0.19 +/- 0.31 (+/+, N = 18) (NS). Changes in LDL-C and HDL-C correlated with the logarithm of the change in free T4 (fT4), expressed as the fT4 posttreatment/fT4 pretreatment ratio (r = -0.81, P < 0.001; and r = -0.62, P < 0.001, respectively). In conclusion, in the transition from hypo- or hyperthyroidism to euthyroidism, no association is found between AvaII genotype and changes in plasma LDL-C nor between TaqIB genotype and changes in HDL-C. Changes in LDL-C and HDL-C correlate with changes in fT4. PMID- 10843166 TI - Primary hyperaldosteronism in essential hypertensives: prevalence, biochemical profile, and molecular biology. AB - There is evidence that primary aldosteronism (PA) may be common in patients with essential hypertension (EH) when determinations of serum aldosterone (SA), plasma renin activity (PRA), and the SA/PRA ratio are used as screening. An inherited form of primary hyperaldosteronism is the glucocorticoid-remediable aldosteronism (GRA) caused by an unequal crossing over between the CYP11B1 and CYP11B2 genes that results in a chimeric gene, which has aldosterone synthase activity regulated by ACTH. The aim of this study was to evaluate the prevalence of PA and the GRA in 305 EH patients and 205 normotensive controls. We measured SA (1-16 ng/dL) and PRA (1-2.5 ng/mL x h) and calculated the SA/PRA ratio in all patients. A SA/PRA ratio level greater than 25 was defined as being elevated. PA was diagnosed in the presence of high SA levels (>16 ng/dL), low PRA levels (<0.5 ng/mL x h), and very high SA/PRA ratio (>50). Probable PA was diagnosed when the SA/PRA ratio was more than 25 but the other criteria were not present. A Fludrocortisone test was done to confirm the diagnosis. GRA was differentiated from other forms of PA by: the aldosterone suppression test with dexamethasone, the high levels of 18-hydroxycortisol, and the genetic detection of the chimeric gene. In EH patients, 29 of 305 (9.5%) had PA, 13 of 29 met all the criteria for PA, and 16 of 29 were initially diagnosed as having a probable PA and confirmed by the fludrocortisone test. Plasma potassium was normal in all patients. The dexamethasone suppression test was positive for GRA in 10 of 29 and 18 hydroxycortisol levels were high in 2 of 29 patients who had also a chimeric gene. In normotensive subjects, 3 of 205 (1.46%) had PA, and 1 of 205 had a GRA. In summary, we found a high frequency of normokalemic PA in EH patients. A high proportion of PA suppressed SA with dexamethasone, but only a few had a chimeric gene or high levels of 18-hydroxycortisol. These results emphasize the need to further investigate EH patients. PMID- 10843167 TI - Insulin-induced increment of coronary flow reserve is not abolished by dexamethasone in healthy young men. AB - Hyperinsulinemia is a risk factor for coronary artery disease. Previous studies have reported that hyperinsulinemia increases cardiac and skeletal muscle sympathetic nerve activity and skeletal muscle blood flow in normal subjects. However, little is known about insulin's effects on myocardial blood flow in humans. The purpose of this study was to investigate whether physiological hyperinsulinemia affects myocardial blood flow and flow reserve in healthy subjects. Additionally, the role of the sympathetic nervous system in regulating insulin's effects on coronary perfusion was tested. We used positron emission tomography and oxygen-15-labeled water to measure myocardial blood flow and coronary flow reserve in 16 healthy nonobese men (age, 34 +/- 4 yr; maximal aerobic capacity, 32 +/- 3 mL x g(-1) x min(-1); blood pressure, 118 +/- 10/65 +/ 8 mm Hg) at fasting and during euglycemic hyperinsulinemic clamp (1 mU x kg(-1) x min(-1) for 80 min). To study the role of the sympathetic nervous system, each subject was studied twice: once after administration of dexamethasone (dexa+) for 2 days (2 mg per day) and once without previous medication (dexa-). All studied subjects had normal left ventricular mass, function, and findings in stress echocardiography. Resting myocardial blood flow was 0.76 +/- 0.19 mL x g(-1) x min(-1), and a significant increase in flow was detected after adenosine infusion (140 microg/kg x min for 5 min i.v.), both in the basal fasting state (P < 0.001) and during hyperinsulinemia (P < 0.001). However, the flow response to adenosine was significantly higher during hyperinsulinemia, thus leading to a higher hyperemic flow (3.38 +/- 0.97 vs. 4.28 +/- 1.57 mL x g(-1) x min(-1), basal vs. hyperinsulinemic, P < 0.01) and higher coronary flow reserve (4.6 +/- 1.2 vs. 5.8 +/- 1.9, respectively, P < 0.05). Pretreatment with dexamethasone did not significantly change the resting blood flow [0.72 +/- 0.22 vs. 0.76 +/- 0.19 mL x g(-1) x min(-1), dexa+ vs. dexa-, not significant (NS)], the adenosine stimulated flow (3.56 +/- 1.49 vs. 3.38 +/- 0.97 mL x g(-1) x min(-1), respectively, NS), or the hyperinsulinemic adenosine-stimulated blood flow (4.68 +/- 1.74 vs. 4.28 +/- 1.57 mL x g(-1) x min(-1), respectively, NS). Coronary flow reserves in the basal state (5.3 +/- 2.7 vs. 4.6 +/- 1.2 mL x g(-1) x min(-1), dexa+ vs. dexa-, NS) and during hyperinsulinemia (6.8 +/- 2.9 vs. 5.8 +/- 1.9 mL x g(-1) x min(-1), respectively, NS) tended to be (but were not) significantly higher after dexamethasone treatment. These results demonstrate that insulin acts as a vasodilatory hormone also in the coronary vasculature. Because the insulin induced increment of myocardial flow reserve remained unchanged by dexamethasone pretreatment, centrally mediated sympathetic activation seems not to play a major role in regulating insulin action on myocardial perfusion in healthy subjects. PMID- 10843168 TI - Continuation of growth hormone (GH) replacement in GH-deficient patients during transition from childhood to adulthood: a two-year placebo-controlled study. AB - Previous studies have demonstrated beneficial effects of GH replacement, in adults with GH deficiency (GHD), on body composition, physical fitness, and quality of life. These studies, however, concern patients with adult-onset GHD or childhood-onset (CO) patients enrolled several years after withdrawal of initial therapy. So far, the effects of continuation of GH-administration in patients with CO-GHD have not been examined. We studied a group of nineteen young adults (13 males + 6 females; 16-26 yr old; mean age, 20.2 +/- 0.65 yr) with CO-GHD, in a randomized, parallel, double-blind, placebo-controlled trial for 1 yr, followed by an open phase with GH for 1 yr. All patients received GH therapy at the start of study, and trial medication (GH/placebo) was given in a similar dose. Patients randomized to continued GH treatment exhibited no significant changes in any parameters tested, but intra- and interindividual variations in insulin-like growth factor (IGF)-I levels could suggest compliance problems. Discontinuation of GH for 1 yr resulted in a decrease in serum IGF-I, from 422.0 +/- 56.8 to 147.8 +/- 33.4 microg/L, in the placebo group (P = 0.003). After discontinuation of GH for 1 yr, an increase in total body fat (TBF, kg), measured by dual-energy x-ray absorptiometry scan, was seen [placebo: 22.7 +/- 2.7 to 26.5 +/- 2.5 (P = 0.01); GH: 16.2 +/- 2.1 to 17.2 +/- 2.1 (not significant)]. Resumption of GH after placebo was followed by increments in serum IGF-I (microg/L) [from 147.8 +/ 33.4 to 452 +/- 76 (P = 0.001)] and IGF-binding protein 3, as well as in fasting glucose (mmol/L) [4.9 +/- 0.2 vs. 5.3 +/- 0.2 (P = 0.03)]. After resumption of GH lean body mass (kg) increased [52.4 +/- 4.9 vs. 60.7 +/- 5.6 (P = 0.006)]. Likewise, resumption of GH therapy increased thigh muscle volume and thigh muscle/fat ratio, as assessed by computed tomography [muscle volume (cm2/10 mm): 118.2 +/- 11.7 vs. 130.0 +/- 10.9 (P = 0.002); muscle/fat ratio: 1.33 +/- 0.24 vs. 1.69 +/- 0.36 (P = 0.02)]. In conclusion, discontinuation of GH treatment in GHD patients, during the transition from childhood to adulthood, induces significant and potentially unfavorable changes in IGF-I and body composition, both of which are reversed after resumption of GH treatment. By contrast, continuation of GH therapy results in unaltered IGF-I and body composition. We recommend continuation of GH therapy in these patients, to be undertaken in collaboration between pediatricians and adult endocrinologists. PMID- 10843169 TI - Trandolapril does not improve insulin sensitivity in patients with hypertension and type 2 diabetes: a double-blind, placebo-controlled crossover trial. AB - Angiotensin-converting enzyme (ACE) inhibitors are increasingly used as first line therapy for hypertension in type 2 diabetes mellitus and are widely believed to improve insulin sensitivity (M). However, the evidence for the latter effect does not stand close scrutiny. We have assessed the effect of the ACE inhibitor trandolapril on M in 16 patients (mean +/- SD age, 58 +/- 10.6 yr) with mild-to moderate essential hypertension (initial blood pressure, 173 +/- 14.5/93 +/- 8.0 mm Hg), obesity (body mass index, 30 +/- 5.4 kg/m2), and impaired glucose intolerance (n = 4) or type 2 diabetes (n = 12) in a double-blind, placebo controlled crossover design. All patients underwent three 3-h euglycemic hyperinsulinemic clamp studies (soluble insulin, 1.5 mU/kg x min) after a 2-week placebo run-in and at the end of two 4-week periods of treatment with 2 mg trandolapril or placebo (2-week washout). M (mean +/- SD) did not change with trandolapril: placebo (run-in), 5.2 +/- 1.98 mg/kg x min; placebo, 5.3 +/- 1.70 mg/kg x min; trandolapril, 5.1 +/- 1.65 mg/kg x min; P = 0.58; 95% confidence intervals, -0.74, 0.43 (trandolapril vs. placebo); 95% power to exclude an 8% increase in M. In conclusion, trandolapril had no clinically relevant effect on M in patients with hypertension and type 2 diabetes. Previous reports of improved M during ACE inhibitor treatment may be attributable to suboptimal study design and/or use of surrogate measures of M. PMID- 10843170 TI - Improved diagnostic methods in the follow-up of medullary thyroid carcinoma by highly specific calcitonin measurements. AB - Calcitonin (CT) is an important tumor marker for medullary thyroid carcinoma (MTC). Recent CT assays chiefly recognize the monomeric form of CT (mCT). It was the objective of this study to examine the consequences of the higher specificity of the assay for interpretation of the postoperative CT values in MTC patients. The postoperative mCT concentration was measured in 214 patients with differentiated thyroid carcinoma (MTC excepted; non-MTC patients) to determine a reference range of mCT in totally thyroidectomized patients. Monomeric CT was also determined with a two-site chemiluminescence immunoassay (Nichols) in 94 healthy subjects and in 68 MTC patients. The mCT concentrations were below the detection limit in all examined completely thyroidectomized non-MTC patients. Basal and stimulated mCT values were also below the detection limit in 32 of the 68 MTC patients. The biochemical and imaging diagnosis of the latter patients did not give any indication of tumor recurrence. We conclude that completely thyroidectomized patients with non-MTC do not show any measurable mCT concentrations. In comparison with an unspecific CT-RIA, the more specific mCT determination by immunoluminometric assay permits a more precise differentiation between postoperative normal and pathological values and an earlier diagnosis of recurrent MTC. PMID- 10843171 TI - Risedronate reverses bone loss in postmenopausal women with low bone mass: results from a multinational, double-blind, placebo-controlled trial. BMD-MN Study Group. AB - Our objective was to investigate the efficacy and tolerability of risedronate in postmenopausal women with low bone mass. Women with a mean lumbar spine T-score of -2 or less (n = 543) received 24 months of placebo or risedronate (2.5 or 5 mg/day). All received calcium (1 g/day). The principal outcome measures were bone mineral density (BMD) at the lumbar spine, femoral neck, and femoral trochanter. At 24 months, lumbar spine BMD increased from baseline by 4% with 5 mg risedronate and 1.4% in the 2.5-mg group, compared with no change with placebo. Efficacy was similar in women who were less than 5 yr and more than 5 yr postmenopausal. At 24 months, risedronate (5 mg) had also increased BMD at the femoral neck and trochanter, whereas BMD decreased in the placebo group. BMD increases were seen at all three sites with risedronate (5 mg) after only 6 months of therapy. Risedronate was well tolerated; upper gastrointestinal adverse events were similar to placebo. We conclude that risedronate (5 mg) increases BMD rapidly and effectively and is well tolerated in postmenopausal women with low bone mass, regardless of time since menopause. PMID- 10843172 TI - Prediction of bone mass change after parathyroidectomy in patients with primary hyperparathyroidism. AB - A major challenge in the management of primary hyperparathyroidism (pHPT) is the decision regarding which patients should undergo parathyroidectomy (PTX), although the Consensus Development Conference of the NIH has proposed guidelines for the indication of surgery. In the present study, changes in bone mineral density (BMD) after PTX were compared between pHPT patients who did and did not meet the NIH criteria, and we further tried to predict the BMD change after PTX from preoperative parameters. The subjects were 44 pHPT patients (30 women and 14 men) who had had successful PTX. Lumbar and radial BMD were measured before and 1 yr after PTX by dual energy x-ray absorptiometry and single photon absorptiometry, respectively. Average annual percent increases in lumbar and radial BMD after PTX were 12.2 +/- 1.4% and 11.6 +/- 1.6% (mean +/- SEM), respectively, and those net increases were 0.0803 +/- 0.0008 and 0.0484 +/- 0.0006 g/cm2, respectively. There were no significant differences in percent or net changes in either radial or lumbar BMD after PTX between the groups divided according to each of the NIH criteria, such as age (> or =50 and <50 yr), serum calcium level (> or =12 and <12 mg/dL) or the existence of urinary stones (presence and absence). On the other hand, when the subjects were divided on the basis of radial BMD (above and below a z-score of -2), the annual percent and net increases in lumbar BMD and percent increase in radial BMD after PTX were significantly higher in the group with the lower z-score. Next, patients were divided into two groups with and without the indication of PTX based on NIH guidelines. Twenty-nine patients had the surgical indication by meeting one or more of these criteria and 15 patients had no indication without meeting any of the criteria. There were no significant differences between the two groups in annual percent or net changes in radial or lumbar BMD after PTX. A stepwise multiple regression analysis revealed that serum alkaline phosphatase level and the severity of cortical bone mass reduction were the best predictors of both percentage and net changes in lumbar BMD, with high determination coefficients (r2 > 0.7). In conclusion, a considerable increase in BMD could be obtained after PTX even in patients without surgical indication from the NIH. Alkaline phosphatase and the severity of cortical bone mass reduction are clinically useful for predicting the changes in lumbar BMD after PTX. The present findings provide a useful clue for the indication of surgery in pHPT. PMID- 10843173 TI - A mutation in the 5' non-high mobility group box region of the SRY gene in patients with Turner syndrome and Y mosaicism. AB - In Ullrich-Turner syndrome (UTS) patients, the presence of a Y-chromosome or Y derived material has been documented in frequencies ranging from 4-61%. Mutations of SRY (testis-determining gene) constitute the cause of XY sex reversal in approximately 10-15% of females with pure gonadal dysgenesis. Most of these mutations have been described in the HMG (high mobility group) box of the gene, which is the region responsible for DNA binding and bending; however, various mutations outside the HMG box have been reported. We carried out molecular studies of the SRY gene in three patients with a UTS phenotype and bilateral streaks; two presented a 45,X/46,XY mosaic, and the third a Y marker chromosome. In two patients a missense mutation, S18N, was identified in the 5' non-HMG box region in DNA from blood and both streaks; this mutation was not identified in 75 normal males. Sequencing of the DNA region of interest was normal in the father and older brother of patient 1, demonstrating that in this patient the mutation was de novo. A previous report of a 46,XY patient with partial gonadal dysgenesis who presented the same mutation as our patients indicates the probable existence of a hot spot in this region of the SRY gene and strengthens the possibility that all gonadal dysgeneses constitute part of a spectrum of the same disorder. It also demonstrates that a single genetic abnormality can result in a wide range of phenotypic expression. PMID- 10843174 TI - Continuation of growth hormone (GH) therapy in GH-deficient patients during transition from childhood to adulthood: impact on insulin sensitivity and substrate metabolism. AB - The appropriate management of GH-deficient patients during transition from childhood to adulthood has not been reported in controlled trials, even though there is evidence to suggest that this phase is associated with specific problems in relation to GH sensitivity. An issue of particular interest is the impact of GH substitution on insulin sensitivity, which normally declines during puberty. We, therefore, evaluated insulin sensitivity (euglycemic glucose clamp) and substrate metabolism in 18 GH-deficient patients (6 females and 12 males; age, 20 +/- 1 yr; body mass index, 25 +/- 1 kg/m2) in a placebo-controlled, parallel study. Measurements were made at baseline, where all patients were on their regular GH replacement, after 12 months of either continued GH (0.018 +/- 0.001 mg/kg day) or placebo, and finally after 12 months of open phase GH therapy (0.016 mg/kg x day). Before study entry GH deficiency was reconfirmed by a stimulation test. During the double-blind phase, insulin sensitivity and fat mass tended to increase in the placebo group [deltaM-value (mg/kg x min), -0.7 +/- 1.1 (GH) vs. 1.3 +/- 0.8 (placebo), P = 0.18; deltaTBF (kg), 0.9 +/- 1.2 (GH) vs. 4.4 +/- 1.6 (placebo), P = 0.1]. Rates of lipid oxidation decreased [delta lipid oxidation (mg/kg x min), 0.02 +/- 0.14 (GH) vs. -0.32 +/- 0.13 (placebo), P < 0.05], whereas glucose oxidation increased in the placebo-treated group (P < 0.05). In the open phase, a decrease in insulin sensitivity was found in the former placebo group, although they lost body fat and increased fat-free mass [M value (mg/kg x min), 5.1 +/- 0.7 (placebo) vs. 3.4 +/- 1.0 (open), P = 0.09]. In the group randomized to continued GH treatment almost all hormonal and metabolic parameters remained unchanged during the study. In conclusion, 1) discontinuation of GH therapy for 1 yr in adolescent patients induces fat accumulation without compromising insulin sensitivity; and 2) the beneficial effects of continued GH treatment on body composition in terms of decrease in fat mass and increase in fat-free mass does not fully balance the direct insulin antagonistic effects. PMID- 10843175 TI - Estrogens exert route- and dose-dependent effects on insulin-like growth factor (IGF)-binding protein-3 and the acid-labile subunit of the IGF ternary complex. AB - We have previously shown that exogenous estrogens exert route-dependent effects on serum GH and insulin-like growth factor I (IGF-I) levels. IGF-I circulates as a ternary complex with IGF-binding protein-3 (IGFBP-3) and the acid-labile subunit (ALS). It is not known whether IGFBP-3 and ALS in blood are regulated by estrogen and, if so, whether this is also route dependent. In the present study we investigate the effects on IGFBP-3 and ALS of oral and transdermal estrogens (study 1), of different oral estrogen formulations (ethinyl estradiol, conjugated estrogen, and estradiol valerate; study 2), of different estrogen dosages (study 3) in normal postmenopausal women, and of oral estrogen in hypogonadal GH deficient women (study 4). Administration of oral, but not transdermal, estrogen in normal postmenopausal women significantly decreased serum levels of IGFBP-3 and ALS (P < or = 0.005). The suppressive effects were similar with different oral estrogen formulations, and the degree of suppression increased with estrogen dosage. In hypogonadal GH-deficient women, oral estrogen treatment also significantly reduced IGFBP-3 and ALS (P = 0.02). The changes in IGF-I in each of the four studies paralleled the changes in both IGFBP-3 and ALS. In conclusion, exogenous estrogens suppress serum IGFBP-3 and ALS in a route- and dose-dependent manner, which are in parallel with the effects on serum IGF-I. These actions of oral estrogen are independent of endogenous GH status. PMID- 10843176 TI - Serial analysis of gene expression as a tool to assess the human thyroid expression profile and to identify novel thyroidal genes. AB - The assessment of the expression profile of normal human thyroid tissue using serial analysis of gene expression (SAGE) generated a collection of 10,994 sequence transcripts (tags). Each tag represented a messenger RNA transcript, and, in total, 6099 different tags could be distinguished. The presence and abundance of thyroid-specific transcripts showed the overall expression profile to be from a normal thyroid cell. The expression level of several transcripts was confirmed on Northern blot. Seventy percent of tags could not be attributed to a known human gene and, therefore, possibly correspond to novel genes putatively involved in thyroid function. The tag sequence generated by the SAGE technique can be used to further characterize these novel genes. In this way, application of the SAGE technique to thyroid tissue gives insight in the expression profile of a normal thyroid gland and provides the information to characterize novel genes involved in thyroid pathology, such as congenital hypothyroidism and thyroid neoplasia. PMID- 10843177 TI - Spatio-temporal expression of the trans-acting splicing factors SF2/ASF and heterogeneous ribonuclear proteins A1/A1B in the myometrium of the pregnant human uterus: a molecular mechanism for regulating regional protein isoform expression in vivo. AB - Many of the human myometrial proteins associated with uterine quiescence and the switch to coordinated contractions at the onset of labor exist as alternatively spliced isoforms. There is now extensive evidence to indicate that the nuclear concentrations of the trans-acting splicing regulators SF2/ASF and hnRNP A1/A1B are fundamental in regulating the expression of specific protein isoforms derived from alternative splicing of single precursor messenger ribonucleic acid transcripts. The question thus arose as to whether these factors were also involved in regulating the expression of specific myometrial protein species within different uterine regions during human gestation and parturition. SF2/ASF and hnRNP A1/A1B expression was therefore determined in paired upper (corpus) and lower segment myometrial samples taken from individual women at term/during spontaneous labor and compared with nonpregnant control samples using specific monoclonal antibodies. We report that SF2/ASF levels were substantially increased in the lower uterine region, and this was associated with a parallel decrease in levels of hnRNP A1/A1B during gestation. Conversely, the opposite pattern was observed within the upper uterine region during pregnancy, where hnRNP A1/A1B was significantly up-regulated and SF2/ASF levels were much less than those found in the lower uterine segment. The differential expression of hnRNP A1/A1B and SF2/ASF in the upper and lower uterine segments may have a primary role in defining the formation of specific myometrial protein species associated with the known contractile and relaxatory properties of these regions before and during parturition. PMID- 10843178 TI - Glucocorticoid stimulation of corticotropin-releasing hormone gene expression requires a cyclic adenosine 3',5'-monophosphate regulatory element in human primary placental cytotrophoblast cells. AB - Production of placental CRH, which is identical to the peptide synthesized and secreted in the hypothalamus, has been linked to human parturition. Glucocorticoids stimulate placental CRH secretion and messenger ribonucleic acid expression, in contrast to their inhibition of CRH synthesis in the hypothalamus. A positive feedforward loop involving glucocorticoid-CRH-ACTH-glucocorticoid is thought to drive the exponential increase in placental CRH leading to delivery. Tissue-specific effects of glucocorticoids on CRH expression are therefore of interest. Using human primary placental cells, we investigated the mechanism by which glucocorticoids stimulate placental CRH gene expression. Nuclear run-on transcription shows that in human placental cells glucocorticoids up-regulate transcription of human CRH (hCRH). Using transient transfection assays we demonstrate that dexamethasone up-regulates both basal and cAMP-stimulated hCRH promoter activity, correlating well with the increase in endogenous CRH peptide levels. Through mutagenesis and deletion analyses we show that dexamethasone stimulation of hCRH gene transcription requires a functional cAMP regulatory element (CRE); this CRE is adequate to confer dexamethasone stimulation upon a heterologous promoter, and electrophoretic mobility shift assay studies show that a placental nuclear protein specifically binds to the hCRH CRE. PMID- 10843179 TI - Expression of leptin receptor in human endometrium and fluctuation during the menstrual cycle. AB - Leptin is secreted by adipocytes and regulates appetite through interaction with hypothalamic leptin receptors (OB-R). Accumulated evidence shows that leptin is involved in the stimulation of reproductive functions and that local expression of leptin and OB-R in the ovary, oocyte, embryo, and placenta plays a role in early development. To investigate the role of leptin in implantation, we examined the expression of OB-R and leptin in the human endometrium. Northern and Western blot analyses and RT-PCR showed that the long form of OB-R (OB-R(L)) messenger ribonucleic acid (mRNA) and protein were expressed. In contrast, leptin mRNA or protein was not detected. All of the splice variants of OB-R (OB-R(T)) and OB R(L) transcripts were expressed in 90% and 84% of the cases, respectively. OB-R mRNA expression peaked in the early secretory phase. Decidual tissue of early gestation also expressed OB-R(T) and OB-R(L). Their incidence and abundance were comparable among endometria with benign uterine diseases and disease-free endometria and were not related to a body mass index within the normal range. The present results indicate that OB-R, but not leptin, is expressed in the human endometrium. PMID- 10843180 TI - Variations in vitamin D-binding protein (group-specific component protein) are associated with fasting plasma insulin levels in Japanese with normal glucose tolerance. AB - The locus of the vitamin D-binding protein (DBP; also known as group-specific component protein or Gc) gene, chromosome 4q12, has been reported to be associated with glucose metabolism in several ethnic groups, including Pima Indians. We have recently reported the association of the DBP genotype with type 2 diabetes mellitus in Japan. The aim of this study was to investigate whether genetic variations of DBP have any influence on glucose metabolism without secondary effects of hyperglycemia or diabetes mellitus using 82 Japanese with normal glucose tolerance. The variations of the DBP gene (Gc 1F, 1S, and 2) were determined by PCR-restriction fragment length polymorphism. Fasting plasma insulin concentration and homeostasis model assessment, an index of insulin resistance, were significantly different based on the DBP genotype (P < 0.01 and P < 0.05, respectively). The people with Gc 1S-2 (5.73 +/- 2.57 microU/mL) and 1S 1S (5.30 +/- 3.46 microU/mL) had significantly higher fasting plasma concentrations than those with 1F-1F (2.84 +/- 1.67 microU/mL) (P < 0.01 and P < 0.03, respectively). There was no significant difference in plasma glucose concentration, body mass index, total cholesterol, triglyceride, and blood pressure. In conclusion, genetic variations of DBP are associated with insulin resistance in Japanese with normal glucose tolerance, which might contribute to the development of type 2 diabetes. PMID- 10843181 TI - Leptin and aging: correlation with endocrine changes in male and female healthy adult populations of different body weights. AB - Aging is associated with changes in plasma levels of several hormones. There are conflicting reports on whether circulating leptin levels change during aging, the possible explanation for which is that alterations in adiposity and body mass index (BMI) also occur. In this study we measured plasma leptin and other hormonal parameters known to influence leptin in 150 men and 320 women of a wide age (18-77 yr) and BMI (18.5-61.1 kg/m2) range. Subjects of each gender were separated into 2 groups of similar BMI, i.e. nonobese (BMI, <30) and obese (BMI, >30), and treated separately. Statistical analysis was performed, treating each group of subjects as a whole population or divided into age groups (<30, 30-50, and >50 yr). BMI-adjusted leptin levels were progressively lower with increasing age in women, with a consistent fall after menopause (-21%; P < 0.001); in men, leptin levels also tended to be lower in subjects more than 50 yr of age, but the reduction was not significant. Multiple linear regression analysis, performed on subjects treated either as a whole population or divided into obese and nonobese, showed that in both genders BMI and age were independent contributors of leptin levels, and there was an inverse relationship between leptin and age in both obese (standardized coefficient beta = -0.25 in women and -0.23 in men; P < 0.01) and nonobese (-0.22 in women and -0.20 in men; P < 0.05) subjects. The correlation of leptin and age with plasma levels of sex and thyroid hormones, GH, insulin-like growth factor I, PRL, and insulin was also evaluated. The variables that correlated with leptin were included in a multiple regression model that included BMI and age. Testosterone in men (-0.43 in nonobese and -0.19 in obese; P < 0.05) and estradiol in women (0.22 in nonobese and 0.24 in obese; P < 0.05) were important contributors to leptin levels; also, dehydroepiandrosterone sulfate in obese women (-0.16) and sex hormone-binding globulin in obese subjects of both genders (0.15 in women and 0.19 in men) were significant determinants in the model. However, none of the hormonal parameters abolished the negative correlation between leptin and age or the gender difference in leptin levels. In conclusion, our data show that in adult humans of different body weight, serum leptin gradually declines during aging; leptin reduction is higher in women than in men, but it is independent from BMI and other age-related endocrine changes. PMID- 10843182 TI - The effects of carbohydrate variation in isocaloric diets on glycogenolysis and gluconeogenesis in healthy men. AB - To evaluate the effect of dietary carbohydrate content on postabsorptive glucose metabolism, we quantified gluconeogenesis and glycogenolysis after 11 days of high carbohydrate (85% carbohydrate), control (44% carbohydrate), and very low carbohydrate (2% carbohydrate) diets in six healthy men. Diets were eucaloric and provided 15% of energy as protein. Postabsorptive glucose production was measured by infusion of [6,6-2H2]glucose, and fractional gluconeogenesis was measured by ingestion of 2H2O. Postabsorptive glucose production rates were 13.0 +/- 0.7, 11.4 +/- 0.4, and 9.7 +/- 0.4 micromol/kg x min after high carbohydrate, control, and very low carbohydrate diets, respectively (P < 0.001 among the three diets). Gluconeogenesis was about 14% higher after the very low carbohydrate diet (6.3 +/ 0.2 micromol/kg x min; P = 0.001) compared to the control diet, but was not different between the high carbohydrate and control diets (5.5 +/- 0.3 vs. 5.5 +/ 0.2 micromol/kg x min). The rates of glycogenolysis were 7.5 +/- 0.5, 5.9 +/- 0.3, and 3.4 +/- 0.3 micromol/kg x min, respectively (P < 0.001 among the three diets). We conclude that under eucaloric conditions in healthy subjects, dietary carbohydrate content affects the rate of postabsorptive glucose production mainly by modulation of glycogenolysis. In contrast, dietary carbohydrate content affects the postabsorptive rate of gluconeogenesis minimally, as evidenced by only a slight increase in gluconeogenesis during severe carbohydrate restriction. PMID- 10843183 TI - A novel A10E homozygous mutation in the HSD3B2 gene causing severe salt-wasting 3beta-hydroxysteroid dehydrogenase deficiency in 46,XX and 46,XY French Canadians: evaluation of gonadal function after puberty. AB - Severe 3beta-hydroxysteroid dehydrogenase (3betaHSD) deficiency is a rare form of congenital adrenal hyperplasia resulting from mutations in the HSD3B2 gene that impair steroidogenesis in both the adrenals and gonads and cause salt-wasting in both sexes and incomplete masculinization of the external genitalia in genetic males. About two thirds of the reported patients are 46,XY. We describe two French-Canadian patients from two families without a known relationship who presented with severe salt-wasting 3betaHSD deficiency in infancy. Although the diagnosis was considered clinically, plasma steroid profiles were confusing. We have thus directly sequenced DNA fragments generated by PCR amplification of the four exons, exon-intron boundaries, and the 5'-flanking regions of the HSD3B2 gene. Sequencing of exon II revealed the presence of a C to A transversion in both alleles of these two cases, thus converting codon 10 (GCA), which codes for Ala, into GAA, encoding Glu. This Ala is highly conserved in the vertebrate 3betaHSD gene family and is located in the putative NAD-binding domain of the enzyme. The mutant type II 3betaHSD enzyme carrying an A10E substitution exhibited no detectable activity in intact transfected Ad293 cells. Both homozygous patients share the same haplotype, spanning approximately 3.3 centimorgans surrounding the HSD3B2 locus, which is consistent with a founder effect for this missense mutation. The 46,XY patient presented with ambiguous genitalia at birth and underwent normal masculinization at puberty, but was azoospermic at 18.5 yr of age. The 46,XX patient presented progressive breast development, menarche, and evidence of progesterone secretion. The only previously reported cases with pubertal follow-up revealed paternity in one male and hypogonadism in one female. These findings demonstrate the complex relationships between the genotype and the gonadal phenotype in severe 3betaHSD deficiency and the difficulty in predicting fertility. PMID- 10843184 TI - Uncoupling protein-2 and -3 messenger ribonucleic acids in adipose tissue and skeletal muscle of healthy males: variability, factors affecting expression, and relation to measures of metabolic rate. AB - Mitochondrial uncoupling protein-2 and -3 (UCP2 and UCP3) may be involved in the modulation of resting metabolic rate and energy balance. To investigate their variability, the influence of this on the variability of energy expenditure, and potential regulatory factors of the expression of the corresponding genes, we measured their messenger ribonucleic acids (mRNAs) in muscle and white adipose tissue of lean, healthy men and correlated the abundance of these mRNAs (attomoles per microg total RNA) with measures of resting metabolic rate, hormone levels (thyroid hormones, insulin, glucagon, leptin, and catecholamines), and fuels potentially involved in energy balance regulation. We also investigated whether the thiazolidinedione, troglitazone, stimulates UCP2 and UCP3 mRNA levels to follow up on the observation that this antidiabetic drug increases the levels of expression in cultured cells. We found UCP2 and UCP3 mRNA levels to be highly variable and poorly correlated with measures of energy expenditure and with most factors affecting energy balance. Only nocturnal urinary norepinephrine excretion could explain a significant fraction of the variability in both UCP2 and UCP3 expression in muscle, but not adipose tissue. Thyroid hormone and norepinephrine excretion were found to contribute to the variability of resting metabolic rate, but this could not be explained by an effect on UCP mRNAs. Troglitazone affected neither the expression of UCPs nor the hormones or the measures of metabolic rate investigated. In conclusion, our results show that the expression of UCP2 and UCP3 genes is quite variable in healthy males and that this variability does not explain that in resting energy expenditure, and suggest that sympathetic activity is an important potential regulator of the expression of these proteins in skeletal muscle. However, the data do not support the concept that regulation of the expression of these genes is the most important level of control of UCP3 and UCP2 functions, and other levels of control have to be invoked. PMID- 10843185 TI - Cytokine gene polymorphisms in autoimmune thyroid disease. AB - Susceptibility to the autoimmune thyroid diseases, Graves' disease (GD) and autoimmune hypothyroidism (AIH), depends on a complex interaction between environmental and genetic factors. The human leukocyte antigen and cytotoxic T lymphocyte-associated-4 regions appear to influence susceptibility to disease, but the effect is not major, and the other genes remain unknown. Cytokines are crucial in the regulation of immune and inflammatory responses and therefore are potential candidate genes for autoimmune thyroid disease. In a case-control study, using a unified method of genotyping, we have examined 15 polymorphisms in 9 cytokine genes in 215 patients with autoimmune thyroid disease (GD, 138; AIH, 77) and 101 normal controls. Polymorphisms in the genes for interleukin-1alpha (IL-1alpha), IL-1beta, IL-1 receptor antagonist, IL-1 receptor 1, IL-4, IL-4 receptor, IL-6, IL-10, and transforming growth factor-beta were investigated. Genotyping was performed using the PCR and sequence-specific primers. Analysis showed a reduced frequency of the variant t allele in the IL-4 promoter polymorphism (position 590) in patients with GD and in the entire patient group (GD and AIH) compared with the control group [corrected P (Pc) = 0.00004 and Pc < 0.00001 for GD and all patients, respectively]. This was reflected in a reduction in the heterozygote genotype in the patient groups compared to the controls [c/t heterozygotes GD, 12%; Pc = 0.06, odds ratio, 0.4 (95% confidence interval, 0.2 0.7); all patients, 11%; Pc = 0.008; odds ratio, 0.4 (95% confidence interval, 0.2-0.7); control subjects, 23%]. There were no significant differences between the study groups for the other polymorphisms examined, and subgroup analysis revealed no association with clinical parameters of disease. These results suggest that an IL-4 variant or a closely linked gene has a modest protective effect against the development of autoimmune thyroid disease, particularly GD. This variation in the IL-4 gene may provide further clues to the pathogenesis of autoimmune thyroid disease and other organ-specific autoimmune diseases. Furthermore, these results suggest that subtle variation in immunoregulatory genes may be associated with autoimmune disease states. PMID- 10843186 TI - Signal transduction characteristics of the corticotropin-releasing hormone receptors in the feto-placental unit. AB - Placentally derived CRH plays a major role in the mechanisms controlling human pregnancy and parturition. In this study, we sought to investigate the signal transduction mechanisms of CRH Type-1 receptors in the feto-placental unit. To clarify the signal transduction components in placenta and fetal membranes, we investigated the expression of G proteins and adenylate cyclase. Using the nonhydrolysable photoreactive analog [alpha-32P] GTP-azidoanilide and peptide antisera raised against G protein alpha-subunits, we studied coupling of CRH receptors to G proteins in both placental and fetal membranes. Treatment of placental membranes with human CRH (100 nM) increased the labeling of Gq, Go, and Gz but not Gi and Gs. Treatment of fetal membranes with human CRH (100 nM) increased the labeling of Go and Gq but not Gi, Gs, and Gz. These results were supported by experiments that showed that CRH failed to activate adenylate cyclase in these tissues, but induced an increase in inositol phosphates instead. These findings provide new insights into the components of the signal transduction machinery in both fetal and placental membranes and suggest that CRH Type-1 receptors can couple to different G proteins in different tissues. The physiological significance of these observations remains to be elucidated. PMID- 10843187 TI - The relationship between leptin concentration and bone metabolism in the human fetus. AB - This study investigates the relationship between leptin and fetal bone metabolism by measuring fetal blood levels of leptin, carboxy-terminal pro-peptide of type I pro-collagen (PICP; a marker of bone formation) and cross-linked carboxy-terminal telopeptide of type I collagen (ICTP; a marker of bone resorption). The median gestational age at the time of sampling was 23 weeks (range, 18-35). There was a positive correlation between leptin concentration and gestational age (r = 0.543, P < 0.001) and a negative correlation between both PICP and ICTP and gestational age (r = -0.592 and r = -0.550, respectively, and P < 0.001 for both). Also, there was a negative correlation between the concentrations of leptin and both PICP (r = -0.260, P = 0.022) and ICTP (r = -0.622, P < 0.001). Using multiple regression analysis, fetal leptin concentration was positively correlated to the gestational age (r = 0.240, P = 0.042) and negatively correlated to ICTP (r = 0.420, P = 0.001). The increase in leptin concentration with gestational age is consistent with adipose tissue development and the subsequent accumulation of fat mass. The negative correlation between fetal leptin and ICTP suggests that leptin may decrease bone resorption with the overall effect of increasing bone mass. Therefore, leptin may play a role in fetal bone metabolism as part of its effect on fetal growth and development. PMID- 10843188 TI - Reduced parathyroid vitamin D receptor messenger ribonucleic acid levels in primary and secondary hyperparathyroidism. AB - Vitamin D, via its receptor (VDR), inhibits the hormone secretion and proliferation of parathyroid cells. Vitamin D deficiency and reduced parathyroid VDR expression has been associated with development of hyperparathyroidism (HPT) secondary to uremia. VDR polymorphisms may influence VDR messenger RNA (mRNA) levels and have been coupled to an increased risk of parathyroid adenoma of primary HPT. VDR mRNA relative to glyceraldehyde-3-phosphate dehydrogenase mRNA levels were determined by RNase protection assay in 42 single parathyroid adenomas of patients with primary HPT, 23 hyperplastic glands of eight patients with uremic HPT, and 15 normal human parathyroid glands. The adenomas and hyperplasias demonstrated similar VDR mRNA levels, which were reduced (42 +/- 2.8% and 44 +/- 4.0%) compared with the normal glands (P < 0.0001). Comparison of parathyroid adenoma with a normal-sized parathyroid gland of the same individual (n = 3 pairs) showed a 20-58% reduction in the tumor. Nodularly enlarged glands represent a more advanced form of secondary HPT and showed greater reduction in the VDR mRNA levels than the diffusely enlarged glands (P < 0.005). The reduced VDR expression is likely to impair the 1,25(OH)2D3-mediated control of parathyroid functions, and to be of importance for the pathogenesis of not only uremic but also primary HPT. Circulating factors like calcium, PTH, and 1,25(OH)2D3 seem to be less likely candidates mediating the decreased VDR gene expression in HPT. PMID- 10843189 TI - The Gly-->Arg972 amino acid polymorphism in insulin receptor substrate-1 affects glucose metabolism in skeletal muscle cells. AB - Molecular scanning of insulin receptor substrate-1 (IRS-1) revealed several amino acid substitutions. The most common IRS-1 variant, a Gly to Arg972 change, is more prevalent among type 2 diabetic patients. In this study we overexpressed wild-type and Arg972IRS-1 variant in L6 skeletal muscle cells and examined the functional consequences of this polymorphism on insulin metabolic signaling. L6 cells expressing Arg972-IRS-1 (L6-Arg972) showed a decrease in insulin-stimulated IRS-1-associated phosphatidylinositol 3-kinase (PI 3-kinase) activity compared with L6 cells expressing wild-type IRS-1 (L6-WT) as a consequence of decreased binding of p85 subunit of PI 3-kinase to IRS-1. L6-Arg972 exhibited a decrease in both basal and insulin-stimulated glucose transport due to a reduction in the amount of both GLUT1 and GLUT4 translocated to the plasma membrane. Both basal and insulin-stimulated Akt phosphorylations were decreased in L6-Arg972 compared with L6-WT. Basal glycogen synthase kinase-3 (GSK-3) activity was increased in L6 Arg972 compared with L6-WT, and insulin-induced inactivation of GSK-3 was also reduced in L6-Arg972. This change was associated with a significant decrease in insulin-stimulated glucose incorporation into glycogen and glycogen synthase activity in L6-Arg972 compared with L6-WT. These results indicate that the Arg972 IRS-1 polymorphism impairs the ability of insulin to stimulate glucose transport, glucose transporter translocation, and glycogen synthesis by affecting the PI 3 kinase/Akt/GSK-3 signaling pathway. The present data indicate that the polymorphism at codon 972 of IRS-1 may contribute to the in vivo insulin resistance observed in carriers of this variant. PMID- 10843190 TI - Peroxisome proliferator-activated receptor-gamma2 P12A and type 2 diabetes in Canadian Oji-Cree. AB - Among the Oji-Cree of northern Ontario, we previously identified a novel variant in the HNF1A gene, namely G319S, that was strongly associated with type 2 diabetes. However, the majority of subjects with diabetes did not have the HNF1A S319 variant, suggesting that there might be other genetic determinants of diabetes susceptibility. In the course of sequencing candidate genes in diabetic subjects who were homozygous for HNF1A G319/G319, we found that some of them had the PPARG A12 variant. After genotyping PPARG in the entire adult Oji-Cree population, we found that: 1) PPARG A12 was strongly associated with type 2 diabetes in women, but not men; 2) among women, the odds of being affected for carriers of PPARG A12 compared with noncarriers was 2.3 (95% confidence interval, 1.4-3.8); and 3) among women, affected carriers of PPARG A12 had a significantly earlier age-of-onset and/or age-at-diagnosis compared with noncarriers. When taken together with the previously reported association of diabetes with HNF1A in both men and women, the gender-specific association with PPARG A12 confirms that type 2 diabetes is etiologically complex in the Oji-Cree and that at least two genes are involved in determining susceptibility to the disease in these people. PMID- 10843191 TI - Detection of binding and blocking autoantibodies to the human sodium-iodide symporter in patients with autoimmune thyroid disease. AB - The sodium iodide symporter (NIS) is a novel autoantigen in autoimmune thyroid disease (ATD). A recent study has described the development of a bioassay for human (h) NIS antibody detection, but this will not detect antibodies that bind the symporter without modulating its activity. Therefore, the establishment of a binding assay is of importance to determine the overall prevalence of hNIS antibodies in ATD patients. An in vitro transcription and translation system was used to produce [35S]-labeled hNIS. The radiolabeled ligand reacted specifically in immunoprecipitation experiments with rabbit antiserum raised against a peptide fragment of hNIS. Subsequently, the reactivity of control and ATD sera to translated [35S]hNIS was determined using RIAs. A significant difference in the frequency of hNIS antibody-positive sera was found when patients with either Graves' disease (GD) or autoimmune hypothyroidism (AH) were compared with normal controls (P = 0.01 and P = 0.03, respectively). Of 49 GD and 29 AH sera tested, 11 (22%) and 7 (24%), respectively, were found to contain hNIS antibodies. Differences were also significant when the antibody-binding indices of the control group of sera were compared with those of both the GD and the AH patient sera (P < 0.0001 and P = 0.001, respectively). In contrast, sera from 10 patients with Addison's disease and 10 patients with vitiligo (without associated ATD) were all negative for antibody reactivity to the symporter. No differences were detected when the antibody binding indices of either the Addison's disease or the vitiligo sera were compared with those of the normal sera group (P = 0.9 and P = 0.6, respectively). Eight of the 11 (73%) GD and 3 of the 7 (43%) AH sera, which were positive for hNIS antibodies in the immunoprecipitation assay, were also found to inhibit iodide uptake in hNIS-transfected CHO-K1 cells, suggesting the existence of antibodies in some serum samples that bind to the symporter without modulating its function. Overall, a significant correlation was found between the iodide uptake inhibition and the binding assays for hNIS antibody detection (r = 0.49, P < 0.0001). In summary, we have developed a specific and quantitative assay for the detection of hNIS binding antibodies in sera of patients with ATD. This system offers the advantage of studying antibody reactivity against conformational epitopes and will be useful in understanding the role of NIS autoreactivity in the pathogenesis of ATD. PMID- 10843192 TI - Expression of pendrin and the Pendred syndrome (PDS) gene in human thyroid tissues. AB - The gene recently cloned that is responsible for the Pendred syndrome (PDS), an autosomal recessive disease characterized by goiter and congenital sensorineural deafness, is mainly expressed in the thyroid gland. Its product, designated pendrin, was shown to transport chloride and iodide. To investigate whether the PDS gene is altered during thyroid tumorigenesis, PDS gene expression and pendrin expression were studied using real-time kinetic quantitative PCR and antipeptide antibodies, respectively, in normal, benign, and malignant human thyroid tissues. The results were then compared to those observed for sodium/iodide symporter (NIS) expression. In normal tissue, pendrin is localized at the apical pole of thyrocytes, and this in contrast to the basolateral location of NIS. Immunostaining for pendrin was heterogeneous both inside and among follicles. In hyperfunctioning adenomas, the PDS messenger ribonucleic acid level was in the normal range, although immunohistochemical analysis showed strong staining in the majority of follicular cells. In hypofunctioning adenomas, mean PDS gene expression was similar to that detected in normal thyroid tissues, but pendrin immunostaining was highly variable. In thyroid carcinomas, PDS gene expression was dramatically decreased, and pendrin immunostaining was low and was positive only in rare tumor cells. This expression profile was similar to that observed for the NIS gene and its protein product. In conclusion, our study demonstrates that pendrin is located at the apical membrane of thyrocytes and that PDS gene expression is decreased in thyroid carcinomas. PMID- 10843193 TI - Male-to-female transsexuals have female neuron numbers in a limbic nucleus. AB - Transsexuals experience themselves as being of the opposite sex, despite having the biological characteristics of one sex. A crucial question resulting from a previous brain study in male-to-female transsexuals was whether the reported difference according to gender identity in the central part of the bed nucleus of the stria terminalis (BSTc) was based on a neuronal difference in the BSTc itself or just a reflection of a difference in vasoactive intestinal polypeptide innervation from the amygdala, which was used as a marker. Therefore, we determined in 42 subjects the number of somatostatin-expressing neurons in the BSTc in relation to sex, sexual orientation, gender identity, and past or present hormonal status. Regardless of sexual orientation, men had almost twice as many somatostatin neurons as women (P < 0.006). The number of neurons in the BSTc of male-to-female transsexuals was similar to that of the females (P = 0.83). In contrast, the neuron number of a female-to-male transsexual was found to be in the male range. Hormone treatment or sex hormone level variations in adulthood did not seem to have influenced BSTc neuron numbers. The present findings of somatostatin neuronal sex differences in the BSTc and its sex reversal in the transsexual brain clearly support the paradigm that in transsexuals sexual differentiation of the brain and genitals may go into opposite directions and point to a neurobiological basis of gender identity disorder. PMID- 10843194 TI - Familial hypercalcemia and hypercalciuria caused by a novel mutation in the cytoplasmic tail of the calcium receptor. AB - Familial hyperparathyroidism (HPT), characterized by hypercalcemia and hypercalciuria, and familial benign hypocalciuric hypercalcemia (FHH) are the most common causes of hereditary hypercalcemia. The calcium-sensing receptor (CaR) regulates PTH secretion and renal calcium excretion. Heterozygous inactivating mutations of the gene cause FHH, whereas CaR gene mutations have not been demonstrated in HPT. In a kindred with 20 affected individuals, the hypercalcemic disorder segregated with inappropriately higher serum PTH and magnesium levels and urinary calcium levels than in unaffected members. Subtotal parathyroidectomy revealed parathyroid gland hyperplasia/adenoma and corrected the biochemical signs of the disorder in seven of nine individuals. Linkage analysis mapped the condition to markers flanking the CaR gene on chromosome 3q. Sequence analysis revealed a mutation changing phenylalanine to leucine at codon 881 of the CaR gene, representing the first identified point mutation located within the cytoplasmic tail of the CaR. A construct of the mutant receptor (F881L) was expressed in human embryonic kidney cells (HEK 293), and demonstrated a right-shifted dose-response relationship between the extracellular and intracellular calcium concentrations. The hypercalcemic disorder of the present family is caused by an inactivating point mutation in the cytoplasmic tail of the CaR and displays clinical characteristics atypical of FHH and primary HPT. PMID- 10843195 TI - Pediatric adrenocortical tumors: molecular events leading to insulin-like growth factor II gene overexpression. AB - It has been previously shown that adrenocortical tumors (ACT) in adults exhibit structural abnormalities in tumor DNA in approximately 30% of cases. These abnormalities involve chromosome 11p15 and include loss of heterozygosity, paternal isodisomy, and overexpression of the gene for insulin-like growth factor II (IGF2), correlating with DNA demethylation at this locus. It has been hypothesized that these events occur late in the tumorigenic process in adults and seem to correlate with a worse prognosis. We present 4 pediatric cases of ACT diagnosed at 2.5 yr, 10 months, 12 yr, and 2.2 yr. All 4 patients presented with virilization, and 1 patient also showed signs and symptoms of glucocorticoid excess. The youngest patient's maternal aunt had surgical excision of a more than 15-cm ACT 18 yr previously, but the aunt is doing well at age 23 yr. They all had surgical removal of their tumors. The 2.5-yr-old child also received chemotherapy and radiotherapy because of capsular rupture and, after 3 local recurrences, died 3.3 yr after initial presentation. We investigated all 4 tumors for chromosome 11 structural abnormalities (11p15.5 to 11q23), IGF2 and H19 expression by competitive RT-PCR analysis, and IGF2 methylation patterns by Southern analysis. All 4 tumors (100%) showed a combination of structural abnormalities at the 11p15 locus with mosaic loss of heterozygosity involving 11p. All tumors also had significantly increased IGF2 messenger ribonucleic acid levels relative to normal adrenal (up to 36-fold) and significant IGF2 demethylation (mean, 87%). H19 messenger ribonucleic acid levels were undetectable in 3 of 4 tumors, explained in part by mosaic loss of the actively expressed maternal allele for this imprinted gene. By immunohistochemistry we were able to confirm increased IGF-II peptide levels within the tumor tissue in 10 pediatric patients, including the 4 patients described above. Concomitantly, we also observed nuclear accumulation of p53, suggesting somatic mutations. For the 10-month-old patient, sequencing revealed a p53 germline mutation. We therefore conclude that in pediatric ACT, structural abnormalities of tumor DNA and IGF2 overexpression as well as p53 mutations are very common and are therefore less useful for prognosis than in adults. Our findings support the theory that pediatric ACT, whose IGF2 expression and steroidogenesis evoke the phenotype of the fetal adrenal cortex, may arise because of defective apoptosis. PMID- 10843196 TI - Estradiol acts as a germ cell survival factor in the human testis in vitro. AB - The necessity of estrogens for male fertility was recently discovered in studies on both estrogen receptor alpha knockout and aromatase (cyp 19 gene) knockout mice. However, direct testicular effects of estrogens in male reproduction have remained unclear. Here we studied the protein expression of ERalpha and the recently described estrogen receptor beta in the human seminiferous epithelium and evaluated the role of 17beta-estradiol, the main physiological estrogen, in male germ cell survival. Interestingly, both estrogen receptors alpha and beta were found in early meiotic spermatocytes and elongating spermatids of the human testis. Furthermore, low concentrations of 17beta-estradiol (10(-9) and 10(-10) mol/L) effectively inhibited male germ cell apoptosis, which was induced in vitro by incubating segments of human seminiferous tubules without survival factors (i.e. serum and hormones). Dihydrotestosterone, which, in addition to estradiol, is an end metabolite of testosterone, was also capable of inhibiting testicular apoptosis, but at a far higher concentration (10(-7) mol/L) than estradiol. Thus, estradiol appears to be a potent germ cell survival factor in the human testis. The novel findings of the present study together with the previously reported indirect effects of estrogens on male germ cells indicate the importance of estrogens for the normal function of the testis. PMID- 10843197 TI - Outcome of radiotherapy for acromegaly using normalization of insulin-like growth factor I to define cure. AB - Radiation therapy (RT) has traditionally been considered a useful additional therapy for patients with acromegaly not achieving biochemical remission after surgery. However, recent evidence has suggested that RT is not curative in most patients with acromegaly when normalization of the serum insulin-like growth factor I (IGF-I) level is used to define remission. Therefore, we evaluated the success of RT based on IGF-I level in the 47 patients who received RT as part of their treatment from the cohort of 161 patients with acromegaly seen by us between 1981 and 1999. Four patients in whom no post-RT IGF-I level was available were excluded from the analysis. Of the remaining 43 patients, 32 patients received external beam RT, 6 received fractionated stereotactic radiosurgery, 4 received gamma-knife RT, and 1 received proton beam RT. The most recent IGF-I levels in these 43 patients, obtained a mean of 5.2 yr post-RT (range, 0.8-13.2 yr), were compared to age-adjusted normal ranges. IGF-I levels were normal in 17 patients (39.5%) without the addition of medical therapy. The percentage of patients with a normal IGF-I level generally increased with time post-RT; 27% of patients less than 6 yr post-RT, but 69.2% of patients 6 yr or more post-RT had normal IGF-I levels. Using the more traditional criterion for cure, a random GH measurement, 74% of patients had a GH level below 5 ng/mL, and 44% had a GH level below 2.5 ng/mL and would have been considered in remission based on these criteria. We conclude that with time RT remains a useful adjunctive treatment for many patients with acromegaly. RT should be considered along with appropriate medical therapy in selected patients who do not achieve normalization of IGF-I level after surgery or for those resistant to medical therapy. PMID- 10843198 TI - Mannheim classification of nongenomically initiated (rapid) steroid action(s). AB - There is increasing evidence for rapid effects of steroids that are incompatible with the classical model of genomic steroid action. To address the diversity of mechanisms for rapid steroid signaling described over the past years, a classification of rapid steroid effects has been proposed to promote the discussion and understanding of nongenomic steroid action. PMID- 10843200 TI - Comment on extrapituitary parasellar macroadenoma in Cushing's disease. PMID- 10843199 TI - Five patients with biochemical and/or clinical generalized glucocorticoid resistance without alterations in the glucocorticoid receptor gene. AB - Cortisol resistance (CR) is a rare disease characterized by a generalized reduced sensitivity of end-organs to the actions of glucocorticoids (GCs). GC effects are mediated by the GC receptor (GR). The molecular alterations in CR described thus far were located in the hormone-binding domain of the GR gene. Recent reports of a considerable prevalence of abnormalities in the GR in patients attending the endocrine clinic prompted us to carry out further investigations with respect to GR protein and GR gene in patients attending the endocrine clinic for a broad spectrum of complaints and biochemical evidence suggesting a CR. In the present study, we describe five patients with biochemical and clinical CR. All patients showed a diurnal rhythm of serum cortisol concentrations (albeit at a high level), an insufficient suppression of serum cortisol concentration in reaction to 1 mg dexamethasone (DEX), and variable degrees of androgen overproduction, in the absence of clinical signs and symptoms of Cushing's syndrome. Three of the four female patients presented with complaints of androgen overproduction, two of them in combination with fatigue. The other female patient had severe steroid resistant asthma. The only male patient and his son were asymptomatic. In four patients, we investigated receptor protein characteristics on mononuclear leukocytes in a whole cell DEX binding assay and studied the ability of DEX to inhibit mitogen-induced cell proliferation in mononuclear leukocytes in vitro. In all patients investigated, we found alterations in receptor number or ligand affinity and/or the ability of DEX to inhibit mitogen-induced cell proliferation. To investigate the molecular defects leading to the clinical and biochemical pictures in these patients, we screened the GR gene using PCR/single-strand conformational polymorphism/sequence analysis. No GR gene alterations were found in these patients. In conclusion, the five patients described had clinical and biochemical evidence of CR, but no abnormalities were demonstrated in the GR gene. Probably, as yet undefined alterations somewhere in the cascade of events starting with ligand binding to the GR protein, and finally resulting in the regulation of the expression of GC responsive genes, or postreceptor defects or interactions with other nuclear factors form the pathophysiologic basis of CR in these patients. PMID- 10843201 TI - Follow-up in patients with differentiated thyroid carcinoma with positive 18F fluoro-2-deoxy-D-glucose-positron emission tomography results, elevated thyroglobulin levels, and negative high-dose 131I posttreatment whole body scans. PMID- 10843202 TI - Anti-AIDS agents part 41: synthesis and anti-HIV activity of 3',4'-di-o-(-) camphanoyl-(+)-cis-khellactone (DCK) lactam analogues. AB - DCK lactam analogues were synthesized and evaluated for anti-HIV activity against HIV-1 replication in H9 lymphocyte cells. 4-Methyl-DCK lactam (11a) exhibited potent anti-HIV activity with EC50 and therapeutic index values of 0.00024 microM and 119,333, respectively. PMID- 10843203 TI - Solid-phase/solution-phase combinatorial synthesis of neuroimmunophilin ligands. AB - A novel solid-phase/solution-phase strategy for the synthesis of neuroimmunophilin ligands based on GPI 1046 was developed. The synthesis employs a solid-phase esterification strategy followed by a solution-phase pyruvic amide formation to produce multi-milligram quantities of discrete compounds for assay. The protocol was applied to a production library of 880 discrete compounds. A highlight of the strategy is an aqueous extractive purification of the final compounds using a novel liquid/ice extraction system developed for high throughput. PMID- 10843205 TI - A rationally designed anticancer drug targeting a unique binding cavity of tubulin. AB - A novel mono-THF containing synthetic anticancer drug (WHI-261) was designed for targeting a previously unrecognized unique narrow binding cavity on the surface of tubulin. The anti-cancer activity of WHI-261 was confirmed using MTT assays. The structure-based design, synthesis, and biological activity of WHI-261 are reported. PMID- 10843204 TI - Synthesis of glycolipid analogues that disrupt binding of HIV-1 gp120 to galactosylceramide. AB - HIV-1 has been shown to infect CD4 negative cells by the binding of HIV gp120 to the glycolipid galactosylceramide (1) (GalCer). Several analogues of 1 were prepared to investigate the specific orientation of 1 in the membrane bilayer that is involved in gp120 binding. Interestingly, N-stearyl-1-deoxynojirimycin (8) displayed potent and specific affinity for gp120 equal to that of 1, a finding that may shed light on the antiviral activity of N-butyl-1 deoxynojirimycin. PMID- 10843207 TI - The first synthesis of clausenamine-A and cytotoxic activities of three biscarbazole analogues against cancer cells. AB - Clausemine-A (3), isolated from the stem and root bark of Clausena excavata, was synthesized using Suzuki cross-coupling and Oxidative coupling as the key step. Compound 3, and the other two structurally related biscarbazoles 1 and 2, showed potent cytotoxic activities against a variety of human cancer cell lines in vitro. PMID- 10843206 TI - A new class of highly cytotoxic diketopiperazines. AB - The discovery of a novel class of diketopiperazines possessing potent cytotoxic activity is described. PMID- 10843208 TI - Synthesis and evaluation of antiproliferative activity of a geldanamycin Herceptin immunoconjugate. AB - Geldanamycin was modified with 1,4-diaminobutane to introduce a primary amine that was subsequently employed to provide a maleimide for protein linkage. Monoclonal antibody Herceptin was then derivatized to generate thiol groups that reacted with the maleimide derivative to produce the immunoconjugate. The product showed antiproliferative activity greater than native Herceptin. PMID- 10843209 TI - Structure-activity relationships of halichondrin B analogues: modifications at C.30-C.38. AB - Structurally simplified analogues of halichondrin B were prepared by total synthesis and found to retain potent cell growth inhibitory activity in vitro. PMID- 10843210 TI - Solid-phase parallel synthesis of azarene pyrrolidinones as factor Xa inhibitors. AB - A focused library (4 x 14) prepared from 4-aminopyridine and 4-, 5-, and 6 azoindole templates was synthesized using 14 polymer supported 4-amido-2,3,5,6 tetrafluorophenyl (TFP) sulfonate esters inputs. Several compounds were identified as factor Xa inhibitors (IC50< or =0.1 microM) helping to establish the SAR among these four series of azarene pyrrolidinones. PMID- 10843211 TI - Structure-activity relationship studies of flavopiridol analogues. AB - Cyclin dependent kinases (CDKs) along with the complementary cyclins form key regulatory checkpoint controls on the cell cycle. Flavopiridol is a synthetic flavone that shows potent and selective cyclin-dependent kinase inhibitory activity. In this paper, we report modifications of the 3-hydroxy-1 methylpiperidinyl (D ring) of flavopiridol and their effect on CDK inhibitory activity. PMID- 10843212 TI - Antimycobacterial activity of ionic fullerene derivatives. AB - Positively charged fullerene derivatives, moderately soluble in water:DMSO 9:1, have been tested using three strains of Mycobacterium spp. Some compounds inhibit the growth of Mycobacterium tuberculosis, a human clinical isolate, particularly virulent and resistant, at doses as low as 5 microg/mL. PMID- 10843213 TI - Synthesis of a potent aminocyclitol alpha-mannosidase inhibitor, 1L-(1,2,3,5/4)-5 amino-4-O-methyl-1,2,3,4-cyclopentanetetrol. AB - Demethylthio-, de-S-methyl-, and ethylthio-derivatives of the alpha-mannosidase inhibitor, mannostatin A, have been synthesized and evaluated for their inhibition of Jack bean alpha-mannosidase in order to elucidate the roles of the methylthio group. All derivatives had lowered inhibitory potentials. However, a mannostatin A analogue with a methoxyl instead of the methylthio, exhibited about 2-fold enhancement of the activity, indicating an importance for the methyl group rather than the sulfur atom. PMID- 10843214 TI - Immunostimulatory activity of CpG containing phosphorothioate oligodeoxynucleotide is modulated by modification of a single deoxynucleoside. AB - Phosphorothioate oligodeoxynucleotides (PS-oligos) containing the CpG motif have immunostimulatory properties. Our earlier study had shown that the immunostimulatory activity of PS-oligos containing the CpG motif can be modulated by incorporation of 2'-O-methylribonucleosides (Zhao, Q.; Yu, D.; Agrawal, S. Bioorg. Med. Chem. Lett. 1999, 9, 3453). Here we show that the immunostimulatory activity of a PS-oligo containing a CpG motif can be modulated by substitution of a single deoxynucleoside at specific sites with either 2'-O-methylribonucleoside or 3'-O-methylribonucleoside in the flanking region to CpG motif. Furthermore, substitution of deoxynucleosides with 2'-O-methoxyethoxyribonucleosides also results in modulating immunostimulatory activity of PS-oligos. PMID- 10843215 TI - Anomalous temperature dependence of peptide films at air-water interface. AB - The tetrapeptide derivative Tyr-Gly-Phe-Ala-OBz (1) forms monolayers as confirmed by compressibility studies carried out at various temperatures. Peptide 1 monolayer exhibits an anomalous structural transition at 40 degrees C as evidenced by pi-A isotherms recorded at different temperatures. The structural transition is also observed in aqueous solution of trifluoroacetate of peptide 1 as evidenced by fluorescence and Raman scattering intensity measurements. PMID- 10843216 TI - Synthesis and structural analysis of the active enantiomer of famoxadone, a potent inhibitor of cytochrome bc1. AB - Famoxadone is a newly commercialized fungicide and potent Qo-site inhibitor of cytochrome bc1. The S-(-)-enantiomer of famoxadone (the active component) was synthesized by two routes and was analyzed computationally and by X-ray crystallography. The molecule displays an extended conformation with flexibility in the structure imparted by the two terminal phenyl groups. In the crystal lattice, intermolecular hydrogen bonds occur between the NH and the oxygen atoms of the heterocycle. PMID- 10843217 TI - Synthesis and structure-activity relationship of novel pyridyl ethers for the nicotinic acetylcholine receptor. AB - The preparation of novel pyridyl ethers as ligands for the nicotinic acetylcholine receptor (nAChR) is described. Variations of the ring size of the azacycle and substitution on the pyridine had dramatic effects on receptor binding affinity with IC50s at the alpha4beta2 nAChR ranging from 22 to >10,000 nM. The most potent molecule was (R)-2-chloro-3-(4-cyanophenyl)-5-((3 pyrrolidinyl)oxy)pyridine 27f with an IC50 of 22 nM. PMID- 10843218 TI - Synthesis of FF-MAS from lithocholic acid. AB - An effective synthesis of 4,4 dimethyl-cholest-8,14,24-trien-3beta-ol (FF-MAS) from lithocholic acid is described, utilising a double oxidation and regioselective Wittig reaction as key steps. PMID- 10843219 TI - Bicyclic pyridones as potent, efficacious and orally bioavailable thrombin inhibitors. AB - A new class of conformationally constrained thrombin inhibitors is described. These compounds contain a unique bicyclic pyridone scaffold which serves as a P3P2 dipeptide surrogate. The synthesis and antithrombotic activity of these inhibitors is reported. PMID- 10843220 TI - Activation of antibacterial prodrugs by peptide deformylase. AB - 5'-Dipeptidyl derivatives of 5-fluorodeoxyuridine (FdU) (1a-d) were synthesized. These compounds are biologically inactive but can be activated by peptide deformylase, which removes the N-terminal formyl group of the dipeptide, to release the active drug FdU via an intramolecular cyclization reaction. Because the deformylase is ubiquitous among bacteria but absent in mammalian cells, 1a-d provide a novel class of potential antibacterial agents. PMID- 10843221 TI - Examination of N-hydroxylation as a prerequisite mechanism of nitric oxide synthase inactivation. AB - L-N5-(1-Hydroxyiminoethyl)-ornithine (L-NHIO) and L-N6-(1-hydroxyiminoethyl) lysine (L-NHIL) were synthesized and tested as potential intermediates in the mechanism-based inactivation of nitric oxide synthase (NOS) by L-N5 iminoethylornithine (L-NIO) and L-N6-iminoethyllysine (L-NIL). Although these compounds were determined to be competitive inhibitors, mechanism-based inactivation was not observed. PMID- 10843222 TI - Novel alpha-glucosidase inhibitors with a tetrachlorophthalimide skeleton. AB - Novel alpha-glucosidase inhibitors with a tetrachlorophthalimide skeleton were prepared and their structure-activity relationships were analyzed. Among them, N phenyl-4,5,6,7-tetrachlorophthalimide (CPOP: 2) and N-(4-phenylbutyl)-4,5,6,7 tetrachlorophthalimide (CP4P: 6) showed very potent inhibitory activity, being more potent than 1-deoxynojirimycin (dNM: 1). Mechanistic studies revealed that CPOP (2) and CP4P (6) inhibit alpha-glucosidase non-competitively and competitively, respectively. PMID- 10843223 TI - Phosphonamidate and phosphothioate dipeptides as potential inhibitors of VanX. AB - In an effort to prepare novel inhibitors of VanX, N-[(1 aminoethyl)hydroxyphosphinyl]-D-alanine 1 and S-[(aminoethyl)hydroxyphosphinyl] thiolacetic acid 2 were synthesized and evaluated as inhibitors of VanX. Phosphonamidate 1 was shown to be a partial competitive inhibitor of VanX with a Ki of 36+/-3 microM, and phosphothioate 2 was shown not to inhibit VanX. PMID- 10843224 TI - High potent and selective arylpiperazine derivatives as ligands for the 5-HT1A receptor. AB - This paper reports the synthesis and affinities on the 5-HT1A versus the alpha1A receptors of new arylpiperazinylalkylthiothienopyrimidine and thiadiazole derivatives 16-24. Arylpiperazines 16-23 show affinities values in the nanomolar range for the 5-HT1A receptor. The compound 16 is highly potent (Ki 0.26 nM, selectivity 28), the derivatives 20 and 21 are less potent, but highly selective (Ki 9.40 and 5.06 nM, selectivity 207 and 73, respectively). PMID- 10843225 TI - Novel arylpiperazines as selective alpha1-adrenergic receptor antagonists. AB - A novel series of arylpiperazines has been synthesized and identified as antagonists of alpha1a adrenergic receptor (alpha1a-AR) implicated in benign prostatic hyperplasia. These compounds selectively bind to membrane bound alpha1a AR with K(i)s as low as 0.66 nM. As such, these potentially represent a viable treatment for BPH without the side effects associated with known alpha1 adrenergic antagonists. PMID- 10843226 TI - First pharmacophoric hypothesis for 5-HT7 antagonism. AB - In order to make the first contribution to the elucidation of essential structural features for 5-HT7 antagonism, a set of thirty 5-HT7 antagonists were selected from the literature. A pharmacophore model was built using Molecular Modeling studies with Catalyst program. The information contained in this model was validated with new synthesized compounds. PMID- 10843227 TI - Modelling studies on the binding of substrate and inhibitor to the active site of human sorbitol dehydrogenase. AB - This study reports a molecular modelling investigation of human sorbitol dehydrogenase complexed with the substrate sorbitol and the inhibitor WAY135 706 based on the structures of human beta3 alcohol dehydrogenase, human sigma alcohol dehydrogenase and horse liver alcohol dehydrogenase. The tertiary structure of human beta3 alcohol dehydrogenase was used as a template for the construction of the model. The rms positional deviation between the main-chain atoms of the initial and final models of sorbitol dehydrogenase is 1.37 A. Similar residue interactions exist between sorbitol dehydrogenase and both sorbitol and inhibitor. Binding of sorbitol in the substrate-binding site results in interactions with Lys-294, Tyr-50, His-69, Glu-150, and NAD+ while WAY135 706 interacts with Ser-46, Lys-294 and Phe-59. The enzyme-inhibitor interactions revealed by this study will be useful in the design of more specific inhibitors. PMID- 10843228 TI - Diastereoisomers of an 'arsenomethionine'-based structure from Sargassum lacerifolium: the formation of the arsenic-carbon bond in arsenic-containing natural products. AB - Separation and 1H NMR spectra of a pair of arsenic-containing diastereoisomers (1a and 1b) isolated from a brown alga has provided support for their structures (proposed on the basis of NMR spectra of the unseparated mixture). The diastereoisomerism and analogies with nitrogen-containing algal lipids indicated that they were derived from an analogue of methionine in which the dimethylarsinoyl- group had replaced amino. Although S-adenosylmethionine is probably the source of methyl and 5'-deoxyribose-5'-yl groups in arsenic containing natural products, the arsenic-carbon bonds in some compounds might be formed by a process in which arsenic replaces nitrogen in amino-acid synthesis. PMID- 10843229 TI - Synthesis and binding affinities of 4-diarylaminotropanes, a new class of delta opioid agonists. AB - A series of 4-diarylaminotropanes has been prepared. Both endo and exo diastereomeric forms bound to the delta opioid receptor but the endo isomers were more potent and selective versus the mu opioid receptor than the exo isomers. The most potent delta opioid agonist (14) exhibited a delta opioid Ki of 0.2 nM and was 860-fold selective over mu. PMID- 10843230 TI - Cyclopentadienyltricarbonylrheniumbenzazepines: synthesis and binding affinity. AB - Analogues of the benzazepine dopamine D1 receptor antagonist SCH-23390 incorporating the cyclo-pentadienyltricarbonyl-rhenium (CPTR) moiety were synthesized and evaluated pharmacologically. The CPTR derivatives retained affinity (0.3-2.9 nM) and D1 selectivity of the parent compound, supporting their use as neuropharmacological surrogates for 99mTc-labeled SPECT radiopharmaceuticals. PMID- 10843231 TI - Carbonic anhydrase inhibitors: synthesis of N-morpholylthiocarbonylsulfenylamino aromatic/heterocyclic sulfonamides and their interaction with isozymes I, II and IV. AB - Several aromatic/heterocyclic sulfonamides possessing free amino, imino or hydrazino moieties were transformed into the corresponding N morpholylthiocarbonylsulfenyl derivatives, by reaction with N morpholyldithiocarbamate in the presence of oxidizing agents (NaClO or iodine). These compounds showed nanomolar inhibition against three CA isozymes, and interesting in vitro tumor cell growth inhibitory properties, against several leukemia, non-small cell lung, ovarian, melanoma, colon, CNS, renal, prostate and breast cancer cell lines. PMID- 10843232 TI - Phosphate prodrugs of PD154075. AB - In the preparation of phosphate prodrugs of PD154075, several strategies of linking a phosphate group to the indole moiety were studied. A novel linker, p hydroxymethylbenzoyloxymethoxycarbonyl, was discovered to provide the phosphate prodrug of PD154075 (compound 9) with significantly increased aqueous solubility, sufficient stability in aqueous solution and good bio-reconversion in vivo. PMID- 10843233 TI - Syntheses and properties of photoactivatable sugar derivatives designed to probe the sugar-binding site of melibiose permease. AB - Three new photoreactive sugar analogues bearing an azido, a diazonium salt or a diazirine group as the photophore as well as a tritium atom were developed. Two of these new photoactivatable compounds gave excellent preliminary results, with a high affinity for the melibiose permease of Escherichia coli. PMID- 10843234 TI - Syntheses and biological evaluation of novel 2alpha-substituted 1alpha,25 dihydroxyvitamin D3 analogues. AB - Novel 2alpha-substituted 1alpha,25-dihydroxyvitamin D3 analogues were efficiently synthesized and their biological activities were evaluated. 2alpha-Methyl 1alpha,25-dihydroxyvitamin D3 (2), whose unique biological activities were previously reported, was modified to 2alpha-alkyl (ethyl and propyl) and 2alpha hydroxyalkyl (hydroxymethyl, hydroxyethyl, and hydroxypropyl) analogues 3-7 by elongation of the alkyl chain and/or introduction of a terminal hydroxyl group. 2alpha-Hydroxypropyl-1alpha,25-dihydroxyvitamin D3 (7) exhibited an exceptionally potent calcium-regulating effect and a unique activity profile. PMID- 10843236 TI - Templating peptide folding on the surface of a micelle: nucleating the formation of a beta-hairpin. AB - NMR spectroscopy is used to show that a 20-residue beta-hairpin peptide sequence derived from ferredoxin I, with a Pro-Asp two-residue type I turn which is uncommon in beta-hairpins, is unstructured in aqueous solution but shows NOE evidence for partial folding in the presence of sodium dodecylsulphate micelles. The peptide has a number of lysine residues in the N-terminal beta-strand capable of interacting with the micelle surface and templating the partial folding of the hairpin by reducing the entropic cost of ordering the peptide backbone. PMID- 10843235 TI - 4,10-Dihydro-4-oxo-4H-imidazo[1,2-a]indeno[1,2-e]pyrazin-2-carboxylic acid derivatives: highly potent and selective AMPA receptors antagonists with in vivo activity. AB - A novel series of 2-substituted-4,5-dihydro-4-oxo-4H-imidazo[1,2-a]indeno[1,2 e]pyrazine derivatives was synthesised. One of them, 4e-a highly water soluble compound exhibited a nanomolar affinity and demonstrated competitive antagonist properties at the ionotropic AMPA receptors. This compound also displayed potent anticonvulsant properties against electrically or sound-induced convulsions in mice after systemic administration, thus suggesting adequate brain penetration. PMID- 10843237 TI - Reversal of experimental myoglobinuric acute renal failure with bioflavonoids from seeds of grape. AB - Rhabdomyolysis may account for about 10% of all cases of acute renal failure (ARF). This study was performed to explore the protective influence of proanthocyanidins from seeds of grape in an experimental model of myoglobinuric ARF. Rats were injected with 50% glycerol (8 mL/kg, im) followed immediately and daily in the next three days by ip proanthocyanidins (20 mg/kg) or saline. After 96 h rats were sacrificed and kidney morphology, kidney cortex peptidase activities, and malondialdehyde (MDA) content were determined. A moderate renal failure was produced by glycerol injection with blood urea of 31.8+/-11.0 vs. 7.68+/-0.24 m mol/L, and serum creatinine of 153. +/-38.2 vs. 39.6+/-9.0 micromol/L, in glycerol-induced ARF vs. control rats, respectively. Rats that received proanthocyanidins in addition to glycerol had significantly lower (p < 0.01) blood urea and serum creatinine levels compared to those receiving glycerol alone. These functional differences between the glycerol and glycerol plus proanthocianidins groups were also confirmed histologically. Kidney cortex dipeptidylpeptidase IV (DPP IV) activity was not significantly changed in glycerol-induced ARF, however, markedly increased after proanthocyanidins treatment. Kidney cortex malondialdehyde content was found significantly increased in glycerol-induced ARF over control level, and was markedly reduced by proanthocyanidin treatment. Taken together, these results provide strong evidence for the protective role of proanthocyanidins from seeds of grape in glycerol induced ARF. The effect is probably due to the antioxidant activity of proanthocyanidins and to increased expression of kidney cortex DPP IV with effective degradation of TNF-alpha. This may provide therapeutic opportunities of preventing and/or treating myoglobinuric ARF in humans. PMID- 10843238 TI - Human red blood cell hemolysate is a potent mitogen for renal tubular epithelial cells. AB - Conditions associated with intravascular hemolysis are often complicated by acute renal failure which is characterized by renal vasoconstriction and tubular injury. However, the effects of human red blood cell (RBC) hemolysate on renal tubular epithelial cell function have not been well characterized. We therefore measured the effect of distilled water-lysed human RBCs on cultured LLC-PK1 cell function. We found that human RBC hemolysate produced a marked effect to promote LLC-PK1 3H-thymidine uptake that was several-fold higher than that produced by maximal concentrations of several known growth factors. Partial purification of human RBC hemolysate by sequential centrifugation and passage over a column that removes low molecular weight substances each significantly reduced, but did not totally eliminate, the effect of human RBC hemolysate to promote 3H-thymidine uptake. Exposure of LLC-PK1 cells to horse myoglobin also stimulated 3H-thymidine incorporation in LLC-PK1 cells. A reducing agent (ascorbic acid) decreased the effect of horse myoglobin and of human RBC hemolysate to promote LLC-PK1 mitogenesis. Ascorbic acid also decreased the methemoglobin content of human RBC hemolysate. The effect of human RBC hemolysate to increase LLC-PK1 incorporation of 3H-thymidine could also be significantly decreased by either of two inhibitors of tyrosine kinase. These results suggest that there are several components of human RBC hemolysate that promote LLC-PK1 3H-thymidine incorporation. One of these components appears to be methemoglobin that exerts its effect via a tyrosine kinase signal transduction pathway. PMID- 10843239 TI - The combined effect of cyclosporine a and gentamicin on enzymuria in the Sprague Dawley rat. AB - Male Sprague-Dawley rats (8 per group) were administered a single oral dose of cyclosporine A (10, 30 and 50 mg/day) for 5 days or vehicle (corn oil, 1.5 mL/kg) and urinary enzymes excretion was monitored. Only minor changes in enzymuria were observed in the 10 and 30 mg/kg group. However, in the 50 mg/kg group, nephrotoxicity was evident by significant increase in the excretion of N-acetyl beta-D-glucosaminidase (NAG), glutamate dehydrogenase (GDH), and lactate dehydrogenase (LDH on day 2 of treatment. As chemotherapeutic drug interaction with cyclosporine A (CyA) is thought to aggravate its nephrotoxicity, the effect of combined CyA (30 mg/kg) and the antibiotic gentamicin (50 mg/kg) for 5 days was investigated. Gentamicin alone caused a significant enzymuria, whilst co treatment of rats with CyA gave rise to increased changes in enzymuria on days 1 and 2, between the groups receiving gentamicin+vehicle and those receiving CyA+gentamicin. This was particularly marked by significant changes in LDH excretion. In contrast these observed differences were not paralleled by changes in serum creatinine and other functional parameters. Treatment with gentamicin, appears to enhance CyA nephrotoxicity, but only in the first 2 days, after this there was no significant differences between the two groups. Our data suggest that urinary enzyme measurements could serve as a valuable non-invasive means of monitoring renal performance in animals or humans who may be exposed to combination of drugs. CyA is found not to potentiate the nephrotoxic effect of gentamicin in the animal model used in this study. It therefore appears safe to use the combined therapy particularly in the treatment of transplant patients. PMID- 10843240 TI - A prospective study of thromboelastography (TEG) and filter life during continuous veno-venous hemofiltration. AB - Anticoagulants are commonly used to prolong circuit life during continuous hemofiltration. However, a clear correlation between routinely performed blood coagulability tests and circuit life has not been demonstrated. This lack of correlation may derive from the limited ability of such tests to describe the likelihood of in vivo clotting. We hypothesized that thromboelastography (TEG), which derives its variables from a closer reproduction of in vivo coagulation, would significantly correlate with filter life. Accordingly, we conducted a prospective pilot study of the correlation between filter life and TEG-derived variables in 21 hemofilters used in 6 critically ill patients admitted to a tertiary intensive care unit. It involved the performance of TEG during steady state anticoagulation, measurement of circuit life, and of routine coagulation variables. The results showed that the mean circuit life was 20.7+/-4.0 h despite an average aPTT of 67.7+/-12.8 s and a mean heparin dose of 472.5+/-96.2 IU/h. The mean INR was 1.4+/-1 and the mean platelet count was 118+/-16 x 10(3)/mm3. Although several TEG variables correlated with heparin dose (p < 0.03), no correlation was found between any of the routine coagulation variables or any of the TEG variables and circuit life. In conclusion, no significant correlation between TEG derived variables or routinely measured coagulation variables and circuit life could be demonstrated. These findings suggest that such tests are not useful indicators of circuit anticoagulation adequacy and that factors other than blood coagulability may play a role in circuit failure. PMID- 10843241 TI - Chronic renal failure and periodontal disease. AB - In order to define the effects of chronic renal failure (CRF) in the progress of gingival inflammation, we studied 6 patients (4 male, 2 female) with CRF who were on chronic hemodialysis for 4.25 (range 1-15) years. Six healthy individuals, age and sex matched were used as controls. The protocol which we used comprised of two periods (a) a 40-day duration period of preparation and (b) a 28-day duration experimental period. During the (a) period, all subjects went through: (1) therapy of the chronic gingivitis and (2) complete control of dental plaque by oral hygiene. During the experimental period, all subjects were advised to avoid, for at least 21 days, any mechanical or chemical media of oral hygiene and went through photographing, recording of gingival index (GI), recording of plaque index (PII), and the collection and quantification of gingival crevicular fluid (GCF). On the 21st day, root planning and polishing were performed and subjects were advised to carry out oral hygiene. On the 28th day, all previous examinations (GI, PII, GCF) were repeated. In both patients and controls, GI, PII and GCF were increased on 7th, 14th and 21st day, without significant differences between the groups and returned to normal (close to zero point) on the 28th day. There are no significant differences between patients with CRF and normal controls in the evolution of experimental gingivitis. Therefore, chronic uremia has no effect on the defense of periodontal tissue against microbial plaque. PMID- 10843242 TI - Outcome of pregnancies with HELLP syndrome complicated by acute renal failure (1989-1999). AB - HELLP syndrome, a syndrome of hemolysis, elevated liver enzymes and low platelets may occur in pregnancy with pre-eclampsia/eclampsia, and its a significant complication is acute renal failure (ARF). The aim of study was to determine frequency and outcome of HELLP syndrome complicated by ARF. Thirty-nine patients with pregnancy-related ARF were treated between Jan 1, 1989 and Jan 1, 1999. In these patients, the most frequent causes were HELLP syndrome (n = 14; 36%), postpartum hemorrhage (n = 10; 26%), pre-eclampsia/eclampsia (n = 6; 15%) and abruptio placenta (n = 4; 10%). Seven of the patients with HELLP syndrome had impairment of consciousness during hospitalization. Of these patients, coma in 5, stupor in 1, confusion in 1 were diagnosed. Twelve of the patients with HELLP syndrome and 14 of the other patients were treated by dialysis. Mann-Whitney U test and chi2 test(corrected by Yates and Fisher exact) were used for statistical analysis. Although serious clinical findings, with supportive treatment, 12 patients with HELLP syndrome and 21 other patients were fully recovered. One patient both with and without HELLP syndrome could not recovered due to diffuse cortical necrosis. Moreover, one patient with HELLP syndrome and 3 other patients were died. Mortality rate of the patients with HELLP syndrome was not found different from those of the other patients (p = 0.544). The causes of death were cerebral hemorrhage in patient with HELLP syndrome and disseminated intravascular coagulation (n = 1), cerebral emboli (n = 1), adult respiratory distress syndrome (n = 1). Fetal death occurred in 4 patients with HELLP syndrome (28.5%) and 7 other patients (28%), and rates were similar (p > 0.5). Finally, HELLP syndrome was the most frequent cause leading to ARF in pregnancy and their prognosis was not different from those of the other patients. PMID- 10843243 TI - Peritubular capillary flow determines tubulointerstitial disease in idiopathic nephrotic syndrome. AB - The spatial relationship between renal perfusion and nephronal structure was determined in 51 nephrotic patients consisting of 11 patients with steroid sensitive, minimal change (MC) nephrosis, 12 patients with steroid resistant, mesangial proliferative (MesP) nephrosis and without tubulointerstitial fibrosis (TIF), 11 patients with steroid resistant, MesP nephrosis and with low grade TIF and 17 patients with focal segmental glomerulosclerosis (FSGS). The intrarenal hemodynamic study revealed a unique correlation between renal perfusion and nephronal structure. A normal or slight reduction in peritubular capillary flow observed in MC or mild MesP nephrosis correlates with an intact tubulointerstitial structure. A moderate reduction in peritubular capillary flow observed in steroid resistant, MesP nephrosis induces a low incidence of TIF. A severe reduction in peritubular capillary flow denotes a higher incidence of TIF as that observed in nephrosis with FSGS. Thus, it is of notion that the reduction in renal perfusion precedes the development of tubulo-interstitial fibrosis and thereby supports the concept of renal perfusion as a crucial determinant of nephronal structure. PMID- 10843244 TI - Renal failure in vibrio vulnificus infection. AB - Vibrio vulnificus infection with septicemia is a life threatening disease in the immunocompromised hosts. Renal involvement has not been documented. We reported herein 8 patients with V. vulnificus septicemia. All were immunocompromised hosts. Four patients had cirrhosis of the liver, 3 were heavy alcohol drinkers and one had systemic lupus erythematosis. Presenting symptomatology included fever, chills, leg pain and skin rash. Renal failure was observed in 6 patients. Four patients died shortly after admission. Two survived with clinical course of tubular necrosis. Renal failure is therefore common in V. vulnificus infection. This should be brought to attention, and vigorous antibiotic treatment is required. The disease may be confused with leptospirosis, scrub typhus, malaria and other forms of sepsis which also present with renal failure. PMID- 10843245 TI - Interleukin-6 gene expression in peripheral blood mononuclear cells from patients undergoing hemodialysis or continuous ambulatory peritoneal dialysis. AB - To compare the interleukin-6 (IL-6) gene expression in the peripheral blood mononuclear cells (PBMCs) and plasma IL-6 levels in patients undergoing continuous ambulatory peritoneal dialysis (CAPD) with those in patients undergoing hemodialysis. Eleven hemodialysis patients, 10 CAPD patients, 15 non dialyzed patients with end-stage kidney disease (ESKD), and 7 healthy controls were included in this study. PBMCs were collected by differential centrifugation. Plasma IL-6 concentration was measured by enzyme immunoassay. Plasma IL-6 levels were significantly increased in the hemodialysis and CAPD patients as compared with non-dialyzed ESKD patients and normal subjects (p < 0.01). Following hemodialysis, plasma IL-6 levels exceeded those before hemodialysis. No significant difference was found in plasma IL-6 levels in CAPD patients and in hemodialysis patients when blood was drawn before hemodialysis. Low but steady state levels of IL-6 mRNA expression were observed in the non-dialyzed ESKD patients. The expression of IL-6 mRNA in PBMCs was significantly increased in the patients undergoing hemodialysis or CAPD as compared with the non-dialyzed ESKD patients. The PBMC IL-6 mRNA was significantly lower in CAPD patients than in hemodialysis patients (p < 0.01). A significant correlation was found between the plasma concentration of IL-6 and the expression of IL-6 mRNA in PBMCs from patients undergoing hemodialysis or CAPD (p < 0.01). The hemodialysis or CAPD procedure contributed to the increase in PBMC IL-6 mRNA expression and plasma IL 6 concentration. CAPD treatment stimulated the production of IL-6 to a lesser extent than hemodialysis treatment. PMID- 10843246 TI - Cardiac arrhythmias and silent myocardial ischemia during hemodialysis. AB - Cardiac arrhythmias are noted in a significant proportion of chronic renal failure (CRF) patients on hemodialysis (HD), and may contribute to cardiovascular mortality. A number of factors have been implicated in the genesis of these arrhythmias. The role of silent myocardial ischemia (SMI), however, has not been evaluated systematically. We prospectively studied 38 unselected CRF patients on regular HD by continuous Holter monitoring starting 24 hours before HD, lasting through the dialysis session and continued for 20 hours thereafter. The recordings were analyzed for frequency, timing and severity of supraventricular and ventricular arrhythmias and SMI as identified by ST-segment depression. Ventricular arrhythmias during HD were noted in 11 (29%) patients (group I), and were potentially life-threatening (Lown Class III and IVa) in 13%. The remaining 27 patients (group II) had no ventricular arrhythmias during HD. There was no difference in the age, sex ratio, duration of HD, blood pressure, fluctuations in weight, hematocrit, predialysis creatinine, sodium, potassium, calcium or inorganic phosphate levels between patients in the two groups. The number of patients with clinical ischemic heart disease was significantly greater in group I. SMI was noted in 72% and 33% of group I and II patients respectively (p = 0.026). 46% of those with and 25% of those without ST changes during HD developed ventricular arrhythmias during HD. Both SMI and ventricular arrhythmias were noted most frequently during the last hour of dialysis. Hypertension, diabetes mellitus and ischemic heart disease were observed more frequently amongst patients with SMI. Ventricular arrhythmias are detected in a significant proportion of CRF patients on HD. These are probably related to coronary artery disease since silent myocardial ischemia is also noted more frequently during HD in these patients. Further studies incorporating coronary angiography are needed in a larger number of patients to establish a definite causal relationship. PMID- 10843247 TI - Direct peripheral venopuncture: another new choice of temporary vascular access. AB - BACKGROUND: Femoral, subclavian, and internal jugular veins access have been widely used for temporary vascular access for hemodialysis, but their use has been associated with a significant complication rate. We report in three selected hemodialysis patients with the procedure of direct peripheral venopuncture as temporary vascular access to reduce complications. METHODS: We have demonstrated hemodialysis via direct puncture of peripheral veins of the antecubital fossa (cephalic vein in the process of arterial inflow to dialyzer and venous outflow from dialyzer to basilic vein) as temporary vascular access for these patients. RESULTS: Renal function of case 1 and case 2 progress to normal status after several sessions of dialytic therapy as well as quit hemodialysis, and case 3 successfully shifts to peritoneal dialysis following four sessions of dialytic therapy. CONCLUSIONS: We recommend this short-term access contribute a important additional new choice in selected patients with acute, reversible renal failure, obstructive uropathy, initiation of peritoneal dialysis, patients on peritoneal dialysis with peritonitis, or under plasmapheresis therapy. PMID- 10843248 TI - Perinephric hematoma and massive gastrointestinal hemorrhage (GIB) complicating polyarteritis nodosa (PAN). PMID- 10843249 TI - Understanding financing options for PACS implementation. Picture archiving and communication systems. AB - The acquisition of expensive equipment such as picture archiving and communication systems (PACS) becomes increasingly difficult as capital budgets become tighter. Traditional ownership financing options in the form of direct purchase or financing (loan) have several limitations including technology obsolescence, higher fixed pricing, limited options for equipment disposal, and the need to tie up valuable capital. Alternative financing options, in the form of conventional lease and risk sharing arrangements, offer several theoretical advantages including technology obsolescence protection in the form of built-in upgrades, preservation of borrowing power, multiple end-of-term options, and payment flexibility (which can be directly tied to realized productivity and operational efficiency gains). These options are discussed, with emphasis on the acquisition of PACS. PMID- 10843250 TI - Integration of computer-assembled digital images and text data as evidence for the oncological record. AB - Digitally created visual images of pertinent patient data have been integrated with text information to formulate a visual evaluation and summary sheet (VESS) using computer processing. The VESS incorporates images of a patient's physical appearance, radiological images, pharmacokinetic graphs, and text information into a 1-page document of the patient's condition. Thus, computer processing of digital images and other information helps to refine patient data presentation, analysis, interpretation, and communication. This form of data management is especially valuable in oncological research, where clinical trials demand rapid, ongoing assessment of results and analysis of large amounts of data. The VESS is an effective mechanism for monitoring both the progress of individual patients and the endpoints of the overall clinical trial. PMID- 10843251 TI - An open architecture for medical imaging systems. AB - Integration of medical imaging is the goal of many standards. Interfile and Digital Imaging and Communications in Medicine (DICOM) standards have made great strides in this area. The impact of DICOM is greater owing to its wider acceptance. However, the lack of conformance testing makes interoperability difficult, often requiring expensive consulting teams in a mixed-vendor environment. DICOM is difficult to learn and deploy because it does not use modern software paradigms. Many hardware and software advances in computer architectures and communications could greatly enhance diagnostic imaging if they could be quickly incorporated into medical imaging standards. These problems led to consideration of a solution able to take advantage of state-of-the-art technology and to allow diverse implementations while ensuring the accurate interchange of information between health care providers. It is an object oriented system information model, with the common object request broker architecture (CORBA) as the vehicle to ensure interoperability between different implementations of the model. This report presents the model. PMID- 10843252 TI - Content-based retrieval in picture archiving and communication systems. AB - A COntent-Based Retrieval Architecture (COBRA) for picture archiving and communication systems (PACS) is introduced. COBRA improves the diagnosis, research, and training capabilities of PACS systems by adding retrieval by content features to those systems. COBRA is an open architecture based on widely used health care and technology standards. In addition to regular PACS components, COBRA includes additional components to handle representation, storage, and content-based similarity retrieval. Within COBRA, an anatomy classification algorithm is introduced to automatically classify PACS studies based on their anatomy. Such a classification allows the use of different segmentation and image-processing algorithms for different anatomies. COBRA uses primitive retrieval criteria such as color, texture, shape, and more complex criteria including object-based spatial relations and regions of interest. A prototype content-based retrieval system for MR brain images was developed to illustrate the concepts introduced in COBRA. PMID- 10843253 TI - Architecture of an image capable, Web-based, electronic medical record. AB - With each medical center department creating and maintaining its own patient care related data, nursing and house staff may find it confusing to log into all the information systems necessary to achieve a global perspective of the patient's state. The Medical Information Network Database application provides a logically centralized Worldwide Web viewing application for the physically distributed data. In addition to coordinating data displays for histories, laboratories, pathology, radiology, and discharge summaries, the application can be configured to apply rule sets to the data and remind caregivers of follow-up tests or of possible reactions to treatment protocols. The viewing client runs on any HTML 2.0-compliant browser, although certain applet enhancements (notably for viewing radiological images) require a browser with Java abilities. With this "thin client" approach, the application can be configured to coexist with other applications (such as a PACS viewer), thus centralizing information and reducing the overall number of computers in the medical center. PMID- 10843254 TI - A controlled evaluation of tuned-aperture computed tomography applied to digital spot mammography. AB - The purpose of this work was to compare the detection accuracy of 3-dimensional (3D) modalities of tuned-aperture computed tomography (TACT) with that of conventional 2-dimensional (2D) digital spot mammograms. A standardized mammographic phantom was placed beneath cadaveric breast tissues of varying densities. Five radiologists were asked to detect as many objects (specks, fibers, and low-contrast masses) as possible from 90 displays in a controlled and factorially balanced multivariate experiment. Radiographic exposure was varied systematically, and projections were averaged to ensure stochastic comparability. Scores were weighted to eliminate task-specific bias and were analyzed by multivariate analyses of variance. All display modalities based on the linear application of the 3D TACT reconstruction method yielded significantly higher detection scores for all tasks than did conventional 2D digital spot mammography, which served as the scientific control modality. This effect was found to be statistically significant (P < .001) in spite of significant variations between tissues (P < .001), observers (P < .001), and exposures (P < .01). TACT may be a promising alternative or enhancement to conventional 2D digital mammography for tasks well simulated by this experimental design. PMID- 10843255 TI - The Society for Computer Applications in Radiology. PMID- 10843256 TI - Tests of a cognitive-resource-allocation account of the bizarreness effect. AB - The authors examined the role of cognitive-resource allocation in obtaining the often reported memory advantage for bizarre relative to common information by using the secondary-task method in 4 experiments. In Experiments 1 and 2, they investigated the relationship between differences in cognitive resource allocation during comprehension and in recall for common and bizarre information. In Experiments 3 and 4, they investigated the same relationship but measured cognitive-resource allocation during imaging rather than during comprehension. Although bizarre items required more time to comprehend and to image than did common items, the differences were not reliable predictors of the recall differences between item types. Furthermore, analyses of response latencies to a secondary task provided no compelling evidence in support of a cognitive-resource allocation explanation of bizarreness effects. Implications for the development of a comprehensive model of the influence of bizarreness on memory are discussed. PMID- 10843257 TI - Improvement strategies in free-throw shooting and grip-strength tasks. AB - Participants performed a free-throw shooting task and a grip-strength task before and after imagery, nonspecific arousal, or no instructions. Imagery improved performance in the free-throw shooting task, which is assumed to have more cognitive components than the grip-strength task. Imagery did not improve performance in the grip-strength task, which is assumed to have fewer cognitive components than the free-throw task. Nonspecific arousal, on the other hand, improved performance in the grip-strength task but not in the free-throw shooting task. Athletic experience, confidence levels, and gender were correlated with actual performance levels in both tasks, but not with improvement. Results are discussed within the transfer-appropriate processing framework. PMID- 10843258 TI - Centrality preferences in choices among similar options. AB - Three explanations were explored for the finding that people prefer the middle option rather than the extremes when choosing from an array of similar options. In Study 1, 68% chose the middle item from a set of three highlighters and three surveys, whereas 32% chose an item from either end, p < .0001. In Study 2, 71% selected the middle chair from a row of three chairs that were either all empty, or had a backpack occupying either one of the two end chairs, p < .0001. These results support a minimal mental effort principle rather than a preference for symmetry rule. In Study 3, 54.2% recalled more graphic items from the center poster of a 3-poster collage, whereas 31.3% and 14.5% recalled more items from the left and right posters, respectively, p < .004. These findings lend additional support to a focus of attention explanation. PMID- 10843259 TI - The role of context on strategic actions in Mastermind. AB - When inexperienced solvers play consecutive Mastermind games, they begin to use a strategy in which specific hypothetical types are offered in response to specific feedback types, a strategy known as the modal hypothesis strategy. When participants who had learned that strategy were shifted onto Mastermind games whose code format was different from, and easier than, that they had experienced, their use of the strategy was degraded despite their familiarity and experience in dealing with the specific feedback types occurring in the new code. The results suggest that the use of a domain-specific strategy is at least somewhat influenced by the match between the context in which the strategy has been acquired and the context in which it is being put to use. PMID- 10843260 TI - What determines choice of limb for unimanual reaching movements? AB - Currently, relatively little is known about what drives the choice of limb for goal-oriented reaching. Traditionally, the explanation has been tied predominately to motor dominance as manifested in handedness. This article offers data and an argument suggesting that handedness can be modified by attentional (spatial) information. Although motor dominance may be the controlling factor in the programming and execution of reaching movements at the midline and hemispace ipsilateral (same side) to the dominant limb, attentional information alters the programming of movements in what would be contralateral space. The general trend of behavior is characterized by reaching on the same side as the stimulus, in ipsilateral fashion, a phenomenon explained by kinesthetic efficiency and hemispheric bias. PMID- 10843261 TI - Contextual connections within puns: effects on perceived humor and memory. AB - The supposition that perceived humor or cleverness would be enhanced by adding separate content to the alternative meanings of a pun was explored, using Tom Swifties as the research vehicle. Because detecting a punning word relationship might induce arousal, requiring more effort and attentional resources for processing than noting a contextual connection, memory strength for adverbs was also examined using a recognition procedure. Five studies were conducted in which college students were asked to judge 24 Swifties (content counterbalanced across participants) in which presence of pun and context were manipulated. The punning relationship was primarily responsible for perceived humor and cleverness. Presence of a pun also enhanced memory strength (d') as long as the familiarization task focused on humor and cleverness. Coherent context enhanced perceived cleverness inconsistently. It was suggested that a coherent contextual connection may make a greater contribution in more lengthy forms of pun-based word play, such as fables or "shaggy dog" stories. PMID- 10843262 TI - Are effect sizes and confidence levels problems for or solutions to the null hypothesis test? AB - Some have proposed that the null hypothesis significance test, as usually conducted using the t test of the difference between means, is an impediment to progress in psychology. To improve its prospects, using Neyman-Pearson confidence intervals and Cohen's standardized effect sizes, d, is recommended. The purpose of these approaches is to enable us to understand what can appropriately be said about the distances between the means and their reliability. Others have written extensively that these recommended strategies are highly interrelated and use identical information. This essay was written to remind us that the t test, based on the sample--not the true--standard deviation, does not apply solely to distance between means. The t test pertains to a much more ambiguous specification: the difference between samples, including sampling variations of the standard deviation. PMID- 10843263 TI - Effects of concurrent verbalization on a time-critical, dynamic decision-making task. AB - The authors investigated the effects of concurrent verbalization on students' performance on a time-critical, dynamic decision-making task. After training on Fire Chief (a computer microworld that simulates fighting a forest fire; M. M. Omodei & A. J. Wearing, 1993a), 60 research participants were allocated randomly to 1 of 3 experimental conditions: silence, associative verbalization, or procedural verbalization. Participants who verbalized the bases of their decisions (procedural verbalization) performed significantly worse on average than participants in the silence condition. There was a small but non-significant decrement in the performance of participants who verbalized thoughts other than the bases of their decisions while performing the task (associative verbalization). Their average level was between the levels of participants in the silence and procedural-verbalization conditions. PMID- 10843264 TI - Ordinal position learning and remote anticipation. AB - Rats were runway trained on each of two 3-trial series of reward outcomes. The series are labeled XNY and ZNN, for which X represents a trial that was rewarded with Noyes pellets and N represents a trial that ended with no reward. Units of distinctively flavored breakfast cereals served as reward on trials labeled Y and Z. One group (Floor) had each series occur with a correlated runway floor, either smooth and black or rough and white. For a second group (Memory), the floor cue was uncorrelated with series. Animals in both groups learned to approach the goal rapidly on the 1st trials of the 2 series and slowly on the 2nd trials, but only Group Floor learned to differentiate the 3rd trials of the series. These results recommend a view of serial learning that emphasizes the role played by information about the ordinal position of series items. PMID- 10843265 TI - The Quality Enhancement Research Initiative (QUERI): from evidence to action. PMID- 10843266 TI - Reinventing VA health care: systematizing quality improvement and quality innovation. AB - The Veterans Health Administration (VHA) in the US Department of Veterans Affairs (VA) manages the largest fully integrated health care system in the United States. In 1995, the VHA initiated a reinvention effort that included the most radical redesign of VA health care to occur since the veterans health care system was formally established in 1946. The 2 paramount goals of this reinvention effort were to ensure the predictable and consistent provision of high-quality care everywhere in the system and to optimize the value of VA health care. Although still a work in progress, dramatic results have been achieved toward these ends during the past 5 years. This article provides an overview of the veterans health care system, and it highlights selected aspects of the system's reengineering. It also describes various steps that have been taken to better manage performance and to systematize quality improvement and quality innovation. This information provides a global context that should facilitate understanding of the genesis and purposes of the Quality Enhancement Research Initiative that is described in other articles in this issue of Medical Care. PMID- 10843267 TI - Quality Enhancement Research Initiative (QUERI): A collaboration between research and clinical practice. AB - This article provides an overview of the Quality Enhancement Research Initiative (QUERI), an ambitious attempt to develop a data-driven national quality improvement program for the Veterans Health Administration (VHA) that is fully integrated within VHA's Strategic Framework for Quality Management, as discussed elsewhere in this supplement. QUERI is designed to ensure the systematic translation of findings and products (quality tools that promote use of research findings) to promote optimal patient outcomes and system-wide improvements. In developing QUERI, a framework was created to integrate structural elements (organizational characteristics) and process considerations (those actions and action sequences associated with positive change) with outcomes (both at the patient level and at the systems level). In developing this framework, a process for translation of evidence into action was born. The QUERI process depends on having or discovering accurate information about what services are needed, who needs them, how they should be provided, and relevant outcomes and costs. This article describes the 6-step QUERI process and presents an overview of relevant programmatic details, including QUERI's rigorous review process, and VHA's unique qualifications for establishing a national model for quality improvement. PMID- 10843268 TI - Veterans Affairs Quality Enhancement Research Initiative in chronic heart failure. AB - Chronic heart failure (CHF) is a highly prevalent condition associated with serious morbidity, intense levels of health services use, and shortened survival. It is also a condition for which ameliorative therapies exist. The evidence indicates that there is substantial need to change clinical practice and health care delivery for people with CHF and thereby improve their outcomes. The goal of the Veterans Affairs (VA) Quality Enhancement Research Initiative in CHF (CHF QUERI) is to create measurable, rapid, and sustainable improvements in quality of care and health outcomes of veterans with heart failure. This article describes the current state of knowledge and practice in care for people with CHF. Using the framework of the 5 steps of the QUERI process, we point out the gaps in research and practice that must be filled if the CHF QUERI is to achieve its goal. We relate our recommendations for how the VA can put its research and administrative infrastructure to work to fill the gaps. Lessons learned about CHF in the course of the CHF QUERI will be applicable to all people with heart failure and to all health care systems--VA as well as non-VA--that care for them. PMID- 10843269 TI - Department of Veterans Affairs' Quality Enhancement Research Initiative for Diabetes Mellitus. AB - Diabetes is a common disease, which frequently leads to serious, high-cost complications. Estimates show that in fiscal year 1994 (FY94), 12.5% of outpatients in the Veterans Health Administration (VHA) received diabetes specific medications, accounted for almost 25% of all VHA pharmacy costs, had a hospitalization rate 1.6 times that of veterans without diabetes, and made 3.6 million outpatient visits to VA clinics. Research demonstrates that much of the mortality and morbidity associated with diabetes can be prevented, and rigorous evidence-based guidelines have been developed. The short-term objectives of the Quality Enhancement Research Initiative for Diabetes Mellitus (QUERI-DM) are to (1) gather baseline information on how current VHA diabetes care differs from the VHA guidelines, (2) develop an efficient, validated system for monitoring key diabetes quality standards in the VHA, (3) evaluate the effectiveness of current approaches to diabetes care and the success of guideline implementation initiatives, and (4) initiate 2 to 4 large-scale quality improvement projects to enhance adherence to practice guidelines and evaluate their impact on patient outcomes, including quality of life. PMID- 10843270 TI - Quality Enhancement Research Initiative in ischemic heart disease: a quality initiative from the Department of Veterans Affairs. QUERI IHD Executive Committee. AB - Despite the dramatic fall in ischemic heart disease (IHD) mortality rates over the last 3 decades, it remains the number one cause of death in the United States, and one of the most frequent indications for care by the US Department of Veterans Affairs. National practice guidelines have been developed and disseminated both by societies that specialize in cardiology and within the Veterans Health Administration. Despite these efforts, a substantial minority remains of patients with IHD who are not treated with guideline-recommended therapies. The Quality Enhancement Research Initiative in IHD is a Veterans Health Administration-sponsored initiative to address the gap between guideline recommended therapies and actual Department of Veterans Affairs practice. Because guideline development for patients with IHD is relatively mature, the Quality Enhancement Research Initiative in IHD will concentrate on measuring existing practices, implementing interventions, and evaluating outcomes in veterans with IHD. Measurement of existing practices will be evaluated through analyses of existing Veterans Affairs databases developed for the Continuous Improvement in Cardiac Surgery Program, as well as data collected at the Center for the Study of Practice Patterns in veterans with acute myocardial infarction. To measure existing practices in outpatients with IHD, we plan to develop a new database that extracts electronic data from patient laboratory and pharmacy records into a relational database. Interventions to address gaps between guideline recommendations and actual practice will be solicited and implemented at individual medical centers. We plan to emphasize point-of-care electronic reminders as well as online decision support as methods for improving guideline compliance. PMID- 10843272 TI - Quality Enhancement Research Initiative in Mental Health. AB - The Veterans Administration (VA) recently introduced its Quality Enhancement Research Initiative (QUERI) to facilitate the translation of best practices into usual clinical care. The Mental Health QUERI (MHQ) was charged with developing strategic plans for major depressive disorder (MDD) and schizophrenia. Twenty percent or more of VA service users are affected by 1 of these 2 disorders, disorders that often have a devastating impact on affected individuals. Despite the increasing availability of efficacious treatments for each disorder, substantial gaps remain between best practices and routine care. In this context, the MHQ identified steps critical to the success of a sustained process of rapid cycle health care improvement for MDD and schizophrenia, including research initiatives to close gaps in knowledge of best treatment practices, demonstration projects to close gaps in practice and to expand understanding of effective strategies for implementing clinical guidelines, targeted enhancements of the VA information system, and research and dissemination initiatives to increase the availability of resources to support the accelerated incorporation of best practices into routine care. This article presents an overview of the elements in the initial MHQ strategic plans and the rationale behind them. PMID- 10843271 TI - Quality Enhancement Research Initiative for human immunodeficiency virus/acquired immunodeficiency syndrome: framework and plan. HIV-QUERI Executive Committee. AB - The Veterans Health Administration (VHA) sees approximately equal to 17,000 human immunodeficiency virus (HIV)-infected patients each year, which makes it the largest provider of HIV care in the United States. HIV causes chronic progressive disease that leads to early death. Newer combination antiretro viral treatments are effective but expensive and difficult to use. The HIV Quality Enhancement Research Initiative (HIV-QUERI) uses the QUERI process to identify high-risk and high-volume populations (step 1), which includes those already under VHA care for HIV, those who do not know of their infection, and those at risk for HIV. In identifying best practices (step 2), the HIV-QUERI will benefit greatly from existing guidelines for the care of established HIV infection, but gaps in knowledge regarding adherence to medication regimens and cost-effective screening are large. To identify existing practice patterns (step 3), the HIV-QUERI will develop a clean analytic data set based on Immunology Case Registry files and expand it through a survey of veterans. Interventions to improve care (step 4) will include national, regional, and site-specific feedback on performance relative to quality standards, as well as patient-level and provider-level interventions to improve adherence and support medical decision-making. To document that best practices improve outcomes and quality of life (steps 5 and 6), HIV-QUERI will track indicators on an ongoing basis by use of the Immunology Case Registry database and possible future waves of the survey. In addition, we will require that these issues be addressed in evaluations of HIV-QUERI interventions. In the present article, we present these steps within a framework and plan. PMID- 10843273 TI - Department of Veterans Affairs Quality Enhancement Research Initiative for spinal cord injury. AB - Spinal cord injury (SCI) is a lifelong condition, requiring ongoing efforts by multiple disciplines to stabilize, diminish, or prevent impairments; avoid or limit secondary complications; and improve or maintain social role functioning and quality of life for the individual throughout his or her life. There are approximately 200,000 persons with SCI in the United States, of whom roughly 22% are veterans. The estimated national economic impact of SCI is approximately $9.73 billion per year. These figures illustrate why SCI is an important topic for the Department of Veterans Affairs Quality Enhancement Research Initiative (QUERI). The SCI QUERI will identify gaps in knowledge of SCI treatment and management, develop research efforts to address these gaps, identify best practices for care and management of SCI, and assess whether best practices lead to improved outcomes, including health-related quality of life. PMID- 10843274 TI - Quality Enhancement Research Initiative in stroke: prevention, treatment, and rehabilitation. AB - Stroke is the third leading cause of death and a leading cause of adult disability in the United States. Both within and outside of the Veterans Health Administration (VHA), the lack of a systematic approach to stroke prevention and treatment may have contributed to reduced rates of compliance with recommended practices and increased rates of stroke. Gaps in the knowledge base inhibit a systematic approach to high-quality care within the veteran population. Initial recommendations for closing those gaps are proposed. In some cases (eg, systematic anticoagulation management), the VHA is perceived as a leader in applied research; therefore, a systematic national policy for implementing these clinics may significantly reduce stroke rates. In other areas (eg, carotid endarterectomy), databases exist that would help advance quality and outcomes, but short-term studies are necessary to establish their utility. To promote strategic improvement in prevention, treatment, and rehabilitation for veterans who may be at risk or have had a stroke, specific objectives are proposed to (1) identify best practices for the effective delivery of long-term anticoagulation and enhance veterans' access to these services, (2) develop risk-adjusted models for the surgical preventive procedure carotid endarterectomy to understand facility variation in outcomes so practices can be improved, (3) define a systematic acute stroke management system so that high-quality stroke-related care can be generalizable to a variety of VHA settings, and (4) assess the impact of poststroke rehabilitation on risk adjustment and the location of outcomes so as to facilitate the implementation of best rehabilitation practices. PMID- 10843275 TI - Improving the quality of VA care for patients with substance-use disorders: the Quality Enhancement Research Initiative (QUERI) substance abuse module. AB - Substance-use disorders are costly in both human and economic terms and are highly prevalent among patients in the VA Health Care System. The Quality Enhancement Research Initiative (QUERI) Substance Abuse Module (SAM) seeks to enhance identification and management of patients with substance-use disorders seen in primary care and other medical settings; bolster specialized substance abuse treatment practices; improve care for patients with multiple comorbidities; and strengthen treatment for high-risk and underserved substance-abuse patient subgroups. This article describes how the SAM will achieve these aims by following the QUERI process steps and conducting an integrated set of research projects that incorporates literature reviews and meta-analyses, naturalistic and randomized controlled trials of promising treatments, studies of barriers to guideline implementation, and outcome-oriented evaluations of the implementation of practice guidelines. PMID- 10843276 TI - Database and informatics support for QUERI: current systems and future needs. Quality Enhancement Research Initiative. AB - OBJECTIVES: Our primary objective is to provide an overview of database and informatics support for the Department of Veterans Affairs (VA) Quality Enhancement Research Initiative (QUERI). METHODS: We discuss the role of information technology resources in the QUERI process. We also review current VA information systems and specific databases in terms of strengths and weaknesses for addressing the QUERI goals. A synthesis of the issues and strategies for addressing specific data needs are presented by use of examples from 2 of the QUERI disease modules: Diabetes Mellitus and Human Immunodeficiency Virus. Finally, we discuss issues that need to be considered during development of new information systems to address the needs of clinical quality-improvement efforts. CONCLUSIONS: Quality enhancement in VA health care requires coordination and careful planning among clinical, administrative, research, policy, and information technology leaders to ensure that key clinical process and outcome measures are reliably collected in the VA information systems. As the QUERI progresses, data needs will probably shift from addressing data gaps to developing approaches for feedback and evaluation. Continued and enhanced cooperation among all VHA business processes is vital to the success of the QUERI. PMID- 10843277 TI - From understanding health care provider behavior to improving health care: the QUERI framework for quality improvement. Quality Enhancement Research Initiative. AB - Basic science and health care research provide the evidence base for the scientific practice of medicine. Over the past 2 decades, as increasingly refined tools for improving health and health care have been developed, the health care community has attempted to bridge the gap between available tools and actual health care practices. This gap can be bridged only by influencing health care provider behavior. The VA Quality Enhancement Research Initiative (QUERI) is a program designed to systematically translate research findings into better health care practices, and thus better health outcomes for enrolled veterans. Integrating provider behavior research considerations and findings into each step of the QUERI process will enhance the effectiveness of the initiative. This article presents a provider behavior research framework for planning, implementing, and evaluating quality improvement interventions within QUERI. PMID- 10843278 TI - Myoepithelial carcinoma of the salivary glands: a clinicopathologic study of 25 patients. AB - Salivary gland carcinomas displaying exclusively myoepithelial differentiation (myoepithelial carcinoma) are considered rare. Their histopathologic features, immunohistochemical profile, and clinical behavior are not well characterized. The authors reviewed the clinicopathologic features of 25 salivary gland tumors fulfilling two fundamental histologic criteria: unequivocally malignant and exclusively myoepithelial. For most of these, the original diagnosis was malignant mixed tumor. Thirteen men and 12 women aged 24 to 77 years (mean age, 55 yrs) participated in the study, and most presented with a painless mass. The parotid gland was the most common site (n = 15). Tumors ranged from 2.1 to 5.5 cm, arising either in association with a benign mixed tumor (n = 15) or de novo (n = 10). Histologically, all the tumors displayed infiltrative growth and most had a characteristic multinodular architecture with a cellular periphery and central necrotic/myxoid zones. Epithelioid, hyaline, spindle, clear, or mixed cell types were noted with accompanying myxoid and/or hyalinized extracellular matrix. Ten tumors were high grade cytologically and 15 were low grade. The mitotic rate ranged from three to 51 mitoses per 10 high-power fields. Necrosis was present in 15 tumors and perineural and vascular invasion were identified in 11 and four neoplasms respectively. Immunoreactivities included CAM5.2 (89%), AE1:AE3 (100%), 34betaE12 (92%), cytokeratin 7 (21%), cytokeratin 14 (53%), vimentin (100%), S-100 protein (100%), smooth muscle actin (50%), calponin (75%), muscle-specific actin (31%), glial fibrillary acidic protein (31%), carcinoembryonic antigen (0%), and epithelial membrane antigen (21%). Ultrastructural examination of three tumors showed myoepithelial features. Ten patients developed recurrences, mostly multiple. Follow up of 17 patients showed that eight patients (47%) developed metastases (six high grade, two low grade) and five patients (29%) died of disease (four high grade, one low grade) after a mean of 32 months. Two patients were alive with disease (19 and 49 mos). Ten patients (59%) were without any evidence of disease after a mean of 42.2 months. Myoepithelial carcinomas exhibit a wide spectrum of cytomorphologic features and diverse clinical outcomes. As a result of their morphologic heterogeneity, they can be confused easily with many tumors. Myoepithelial carcinomas have been underrecognized in the past, primarily by being lumped under a broader category of "malignant mixed tumor." Awareness of their unique cytoarchitectural patterns and immunohistochemical profile is crucial for accurate identification. PMID- 10843279 TI - Aggressive digital papillary adenocarcinoma (aggressive digital papillary adenoma and adenocarcinoma revisited). AB - In 1987 a clinicopathologic study by the Armed Forces Institute of Pathology (AFIP) of rare sweat gland tumors, termed aggressive digital papillary adenoma and adenocarcinoma, was published. Since that time, the AFIP has continued to collect these tumors for study. Based on additional follow-up data, we think the original classification of these tumors requires revision. Sixty-seven cases of aggressive digital papillary adenoma and adenocarcinoma were studied according to their clinical characteristics and histologic features. Fifty of these were originally diagnosed as adenoma and 17 as adenocarcinoma. Follow up on 45 (67%) of the patients was obtained. None of the clinical or histologic parameters studied were found to be predictive of recurrence or metastasis, indicating that the originally proposed criteria for distinguishing between benign (adenoma) and malignant (adenocarcinoma) do not predict biologic behavior. When primary tumors were treated by subsequent reexcision or amputation, only one recurred (5%), when not so treated, 11 recurred (50%) regardless of the original diagnosis (p <0.05). Metastasis occurred in six (14%) cases and in three cases led to the death of the patient. Three of these metastatic cases had met the earlier criteria for adenoma. Pulmonary metastases were observed in five cases. No effective treatment for widespread metastatic disease has yet been developed. Because histologic features with prognostic significance could not be demonstrated in this retrospective review, we propose that all aggressive digital papillary tumors be designated aggressive digital papillary adenocarcinoma. PMID- 10843280 TI - Primary malignant melanoma of the urethra: a clinicopathologic analysis of 15 cases. AB - The clinical and pathologic features of 15 primary urethral melanomas occurring in patients (nine women and six men) age 44 to 96 years (mean age, 73 yrs) are described. In the men the tumor involved the distal urethra. In eight women it involved the distal urethra, usually the meatus; both the distal and proximal urethra were involved in one woman. The tumors were typically polypoid and ranged from 0.8 to 6 cm (mean, 2.6 cm) in maximum dimension. A vertical growth phase was present in all tumors, with a prominent nodular component in seven of them. A radial growth phase was seen in nine tumors. The depth of invasion ranged from 2 to 17 mm. The tumors had diffuse, nested, storiform, or mixed growth patterns. The neoplastic cells typically had abundant eosinophilic cytoplasm, large nuclei with prominent nucleoli, and brisk mitotic activity. Melanin pigment was seen in 12 tumors but was conspicuous in only six. At the time of diagnosis, 13 tumors were confined to the urethra and two patients had lymph node metastasis. Nine patients died of disease 13 to 56 months after initial diagnosis and treatment, and one patient had a local recurrence at 4 years and subsequently died of sepsis 1 year later. Three patients were alive and well at 11 months, 23 months, and 7 years. One patient died at the time of the initial operation, and one died of a ruptured aortic aneurysm at 3 years without evidence of melanoma at autopsy. Primary malignant melanomas of the urethra, one fifth of which are amelanotic, must be included in the differential diagnosis of a number of primary neoplasms that involve the urethra, including transitional cell carcinoma, sarcomatoid carcinoma, and sarcomas. Conventional prognostic factors, such as depth of invasion or tumor stage, do not seem to play as important a role in predicting survival as the mucosal location and the nodular growth present frequently in these tumors. PMID- 10843281 TI - Minimal uterine serous carcinoma: diagnosis and clinicopathologic correlation. AB - The clinicopathologic features of uterine serous carcinoma (USC) lacking myometrial invasion, including its putative precursor lesion endometrial intraepithelial carcinoma (EIC), have not been studied extensively. Some USCs may prove fatal even when myometrial invasion is apparently absent, whereas others may be cured with surgery alone. Accordingly, the authors studied eight cases of pure EIC (no invasion identified) and 13 superficial serous carcinomas (SSCs) in which invasion was limited to the endometrial stroma to clarify the behavior of these lesions. The review demonstrated that the most important feature in assessing prognosis is the presence or absence of extrauterine disease at presentation. Thirteen of 14 patients (93%) with EIC or SSC confined to the uterus (stage I or IIA) were disease free and one was dead of unrelated causes at 52 months, whereas seven women who presented with extrauterine disease, even if only microscopic, were either dead of disease or alive with recurrences. Accordingly, patients with EIC or SSC must undergo meticulous surgical staging at the time of hysterectomy. Because the distinction between EIC and SSC based on the identification of stromal invasion is difficult and these lesions share a unique pattern of clinical behavior, the authors regard EIC and SSC measuring 1 cm or less as "minimal uterine serous carcinoma." PMID- 10843282 TI - Clinicopathologic study of mass-screened neuroblastoma with special emphasis on untreated observed cases: a possible histologic clue to tumor regression. AB - Spontaneous regression and maturation of neuroblastoma (NB) are well documented and occur frequently in infants, including those detected by mass screening. To seek histologic clues for regression/maturation in mass-screened NB, clinicopathologic features of 12 tumors that were resected after 2 to 18 months of untreated observation were reviewed. Unobserved screened and age-matched unscreened patients were also studied. To evaluate the possible important role of apoptosis, apoptotic cells were detected by in situ deoxyribonucleic acid (DNA) nick end labeling and immunohistochemical stain for activated caspase-3. Nests with a varying degree of reduced cellularity ("less cellular" and "hypocellular" nests) were common in patients younger than 18 months of age, and were rare in older patients. Two characteristic cells, which have not been focused previously, were frequent, especially in the hypocellular nests. One showed amorphic eosinophilic cytoplasm with pyknotic nuclei and the other contained plump cytoplasm with well-maintained nuclei. These cells were also observed in 89% of the unobserved screened NBs and 79% of the age-matched unscreened patients with good outcome, whereas they could not be confirmed in any of the age-matched unscreened NBs with poor outcome. The amorphic and plump cells were negative for activated caspase-3 and in situ DNA nick end labeling. From these results, the authors hypothesize that these cells most likely represent a degenerative process, in either a state before the activation of caspase-3 or a caspase independent form of cell death. The presence of less cellular and hypocellular nests with amorphic/plump cells may serve as one of the important clues in predicting tumor prognosis. PMID- 10843283 TI - Epithelial mesothelioma with deciduoid features: report of four cases. AB - Deciduoid mesothelioma is the designation given to an unusual morphologic variant of epithelial mesothelioma that closely simulates exuberant ectopic decidual reaction. Because all four previously reported cases involved the peritoneum and occurred in young women without a history of asbestos exposure, it was suggested that deciduoid mesothelioma was a subtype of epithelial mesothelioma characterized by its unique morphology, that it affects a distinct patient population, and that it is unrelated etiologically to asbestos. The author reports four cases of mesothelioma with deciduoid features, all of which originated in the pleura. Three of the patients were men and one was a woman. Their ages ranged from 46 to 78 years (mean age, 67 yrs). Two of the patients had a history of asbestos exposure. These findings indicate that this morphologic variant of mesothelioma is not limited to a specific patient population nor is it restricted to the peritoneum. PMID- 10843284 TI - Enterocolic (lymphocytic) phlebitis: a rare cause of intestinal ischemic necrosis: a series of six patients and review of the literature. AB - Vasculitis of the gastrointestinal tract is known to occur as part of a systemic process but also may be present in a localized form involving only the digestive tract. We report the clinical and pathologic findings of six patients with intestinal ischemia and necrosis resulting from localized phlebitis associated with fresh and/or organized thrombosis of intramural mesenteric veins. None of the patients showed clinical or laboratory evidence of systemic vasculitis. In all cases the arteries were not involved in the inflammatory process. Follow up ranged between 2 and 15 years without recurrence necessitating reoperation. This form of intestinal phlebitis is described in the literature under different terms but lymphocytic phlebitis, granulomatous phlebitis, necrotizing phlebitis, and myointimal venous hyperplasia are probably morphologic variants of the same entity. We propose to unify the nomenclature and to use for this unusual clinicopathologic entity only the generic term of enterocolic (lymphocytic) phlebitis. PMID- 10843285 TI - Immunohistochemical expression of WT1 by desmoplastic small round cell tumor: a comparative study with other small round cell tumors. AB - Desmoplastic small round cell tumors (DSRCTs) present a reciprocal chromosomal translocation, t(11;22)(p13;q12), that results in fusion of Ewing's sarcoma and Wilms' tumor (WT1) genes. The authors evaluated 15 DSRCTs and 71 other tumors often considered in the differential diagnosis for immunoreactivity using a polyclonal antibody directed against the WT1 part of the chimeric protein resulting from this translocation. WT1 immunostaining was performed on paraffin material using the WT(C-19) antibody after heat-antigen retrieval. All the DSRCTs (15 of 15, 100%) demonstrated strong WT1 nuclear immunoreactivity. Ten of 14 nephroblastomas (71%) disclosed WT1-positive nuclei in accordance with the staining reported by others, and rare and focal nuclear positivity was detected in two of 17 rhabdomyosarcomas. WT1 immunoreactivity was not observed in Ewing's sarcoma/primitive neuroectodermal tumors (zero of 21, 0%), neuroblastomas (zero of 17, 0%), or rhabdoid tumors of the kidney (zero of two, 0%). In nephroblastoma, differential diagnosis with DSRCT was not difficult: Clinical and morphologic data are not similar for these two entities. The current study validates WT1 immunoreactivity as a useful marker to separate DSRCT from other small round cell tumors. PMID- 10843286 TI - Salivary heterotopia, cysts, and the parathyroid gland: branchial pouch derivatives and remnants. AB - Five cases of periparathyroid salivary heterotopia associated with cysts were studied. The specimens were obtained from three men and two women age 36 to 62 years who underwent surgery for primary hyperparathyroidism (four patients) and thyroid nodule (one patient). The heterotopia-cyst combination occurred with normal and abnormal parathyroid glands (four inferior and one of unknown location). Review of histologic slides of all parathyroid glands excised from 258 patients during a 1-year period at the Mayo Clinic revealed two similar salivary gland-cyst units. Seven more cases featured one or more periparathyroid cysts, five with other nonsalivary-type epithelial accompaniments. One of the latter additionally had a focus of parathyroid cells in the cyst wall, and associated thyroid parenchyma with C cells, and cartilage. PMID- 10843288 TI - Inherited medullary microcarcinoma of the thyroid: a study of 11 cases. AB - The authors report 11 patients with genetically determined medullary microcarcinomas. Nine patients were either children or adolescents and two patients were young adults. The youngest patient was 7 years old and the oldest was 34 years of age (mean age, 15.4 yrs). The preoperative diagnosis was based on family history and elevated serum calcitonin levels. In addition, six patients had RET protooncogene mutations in exons 10, 11, and 16. Two patients who had the RET protooncogene mutations did not have serum calcitonin measurements. Nine patients had bilateral medullary microcarcinomas (<1.0 cm), whereas the two patients with unilateral tumors demonstrated multifocal disease. The principle microscopic differences between these genetically determined medullary microcarcinomas and larger sporadic (>1 cm) medullary carcinomas were the low incidence of stromal desmoplasia and amyloid deposition, the high incidence of C cell hyperplasia, and the low incidence of lymph node metastases. Only one patient, a 34-year-old man, presented with lymph node metastases. All patients remain disease free 11 to 70 months after diagnosis. This small series of thyroid microcarcinomas illustrates the impact molecular diagnostics is having on the management and prognosis of genetically determined medullary carcinoma. PMID- 10843287 TI - CD10 and BCL-6 expression in paraffin sections of normal lymphoid tissue and B cell lymphomas. AB - In this study the authors explored the value of immunostaining for follicular center B-cell markers, BCL-6 and CD10, in paraffin sections as a tool for the differential diagnosis of B-cell lymphomas. The cases studied comprised reactive lymphoid hyperplasia (RLH; n = 19), follicular lymphoma (FL; n = 50), low-grade mucosa-associated lymphoid tissue (MALT) lymphoma (n = 24), mantle cell lymphoma (n = 19), splenic marginal zone lymphoma (n = 13), diffuse large B-cell lymphoma (DLBCL; n = 54), Burkitt's lymphoma (BL; n = 20), nodular lymphocyte predominance Hodgkin's disease (NLPHD; n = 16), and classic Hodgkin's disease (CHD; n = 13). In RLH, CD10 and BCL-6 were expressed almost exclusively by the follicular center cells. In contrast in FL, the expression of CD10 (39/50) and BCL-6 (34/36) was seen in both follicular and interfollicular neoplastic B cells. Marginal zone/MALT lymphomas and mantle cell lymphoma were always negative. In DLBCL the expression was variable for both CD10 (21/54) and BCL-6 (39/47), with some tumors, including cases of transformed follicular lymphoma (9/10), coexpressing CD10 and BCL-6, and others expressing only BCL-6, and a small group expressing neither marker, possibly reflecting the underlying primary pathogenetic events such as the rearrangement of BCL-2 or BCL-6 genes. BL was always both CD10 and BCL-6 positive. In NLPHD the L&H cells expressed BCL-6 (11/13) but not CD10, whereas in CHD BCL-6 expression was seen in half of the cases. This study demonstrates that both CD10 and BCL-6 are reliable markers of follicular center B cell differentiation. CD10 and BCL-6 immunostaining have an important role in differential diagnosis of FL from RLH and other low-grade B-cell lymphomas. The results also suggest that a CD10/BCL-6 expression pattern may be helpful in identifying main subsets of DLBCL. However, additional studies comparing genotype with immunophenotype are required. PMID- 10843289 TI - Lymphovascular invasion as a predictor of disease progression in prostate cancer. AB - The biologic heterogeneity of prostate cancer (PCa) is evident from the large discrepancy between incidence rates and disease progression and tumor-related deaths. One of the challenges in treating patients with PCa lies in developing nomograms to identify patients who might benefit from adjuvant therapies. Lymphovascular invasion (LVI) is among the variables in PCa recommended to be reported by the Cancer Committee of the College of American Pathologists (CAP), yet few studies have evaluated the prognostic significance and prevalence of LVI in PCa. In the present study, whole-mount specimens from 263 patients with pT3N0 PCa treated by radical prostatectomy by a single surgeon were evaluated for the presence, location, and number of foci of LVI. Foci of LVI were identified in 91 patients. In cases with LVI the number of foci ranged from 1 to 40 with the majority of patients having 1 or 2 foci. LVI was found to be a significant predictor of disease progression in univariate analysis (p <0.0001) and was significantly related to Gleason sum (p <0.001), extra prostatic extension (focal vs established; p = 0.033), and seminal vesicle involvement (p <0.001). Furthermore, in multivariate analysis, LVI was a significant independent predictor of disease progression as well (p = 0.0014). These findings support the CAP recommendations and provide merit for the inclusion of LVI in nomograms to predict disease recurrence in PCa. PMID- 10843290 TI - Xanthogranulomas with inconspicuous foam cells and giant cells mimicking malignant melanoma: a clinical, histologic, and immunohistochemical study of three cases. AB - Histiocytic proliferations can mimic melanocytic tumors and vice versa. The authors describe the clinical, histologic, and immunohistochemical findings of three predominantly mononuclear xanthogranulomas that were misdiagnosed as malignant melanoma by experienced pathologists. All lesions occurred in male patients ranging in age from 14 to 75 years. The tumors presented as dermal nodules, two of which were surrounded by an epidermal collarette and were ulcerated focally. The tumors were composed of a mixed population of large epithelioid and plump spindle cells with pink or pale cytoplasm arranged in nests and short fascicles. Occasional mononuclear cells had cytoplasmic vacuolar changes, but none had well-developed foamy cytoplasm. Rare, multinucleated giant cells were present, but they were not of the Touton type. Mitotic figures were found in all lesions. Immunohistochemically, most tumor cells (80%-90%) were strongly positive for CD68 and a minority of cells (10%-15%), located typically at the periphery of the tumor, was positive for factor XIIIa. Two tumors contained rare cells positive for S-100 protein (5% of tumor cells or less). All tumors were completely negative for tyrosinase (T311), gp100 (HMB-45), and Melan A (A103). Giant and foam cell-poor variants of juvenile xanthogranuloma have been reported previously, mainly in young children. Their occurrence in adolescents and adults is underrecognized. Knowledge of this variant is important to avoid misdiagnosing a benign tumor as malignant melanoma. PMID- 10843291 TI - Cytokeratin 7 and 20 expression in cholangiocarcinomas varies along the biliary tract but still differs from that in colorectal carcinoma metastasis. AB - In the liver, the immunostaining of cytokeratins (CK) 7 and 20 has been used to distinguish usual peripheral cholangiocarcinomas (CC) and colorectal carcinoma metastasis (CRM). However, other subtypes of CC are not infrequent and may be particularly difficult to distinguish from CRM by histology and even immunohistochemistry. Therefore, 48 CC from different locations, either peripheral (n = 19), or nonperipheral, that is, from the large intrahepatic bile ducts, the hilum, and the extrahepatic bile ducts (n = 29), and with different cytoarchitectural patterns were tested for CK7 and CK20 and compared with 31 CRM. CC were positive for CK7 and CK20 in 96% and 70%, respectively, whatever the architecture and differentiation of the tumor. The labeling index (LI) of CK7 in CC was always high, whereas it was low or moderate for CK20. CK20-positive phenotype was significantly more frequent in nonperipheral than in peripheral CC (82% vs 47%; p = 0.007). CRM were all positive for CK20 with a high LI, and mostly negative (81%) for CK7. In conclusion, (1) the CK immunoprofile of CC varies according to the location of the tumor in the biliary tract, peripheral CC being more often CK7+/CK20-, and nonperipheral ones CK7+/CK20+; and (2) a decision tree based on CK20 LI and CK7 positivity allows the distinction of CRM and CC, even for the nonperipheral type. PMID- 10843292 TI - Hyalinizing trabecular adenoma and papillary carcinoma of the thyroid gland express different cytokeratin patterns. AB - It has recently been suggested that hyalinizing trabecular adenoma of the thyroid is an encapsulated variant of papillary carcinoma because of certain similarities of their histology, the occasional occurrence of both tumors in the same gland, and their similar pattern of expression of cytokeratins, including staining for cytokeratin 19. To investigate this notion further, we examined immunocytochemically the expression of a series of cytokeratins in 12 hyalinizing trabecular adenomas and six papillary carcinomas. Hyalinizing trabecular adenoma showed no or minimal reactivity for cytokeratin 19, whereas papillary carcinoma was almost always strongly reactive. Also, hyalinizing trabecular adenoma showed no staining for high-molecular-weight (HMW) cytokeratin, whereas papillary carcinoma was strongly positive. Thus, there are different patterns of cytokeratin 19 and HMW cytokeratin expression in hyalinizing trabecular adenoma and papillary carcinoma. The findings do not support the suggestion that hyalinizing trabecular adenoma is an encapsulated variant of papillary carcinoma. PMID- 10843293 TI - A chromosomal abnormality in hyaline vascular Castleman's disease: evidence for clonal proliferation of dysplastic stromal cells. AB - The pathogenesis of the hyaline vascular variant of Castleman's disease is currently unknown; however, vascular and dendritic cell proliferations are common in this disorder. We report a clonal karyotypic abnormality (46,XX,t(1;16) (p11;p11), del(7)(q21q22),del(8)(q12q22)) in 15 of 20 cells obtained after short term stromal cultures of a typical case of hyaline vascular Castleman's disease (HVCD). There was no histologic, immunohistochemical, or genotypic evidence of a clonal lymphoid or plasma cell proliferation supporting origin of this aberration from the stromal component, possibly dendritic cells. We re-examined 15 previous cases of HVCD and identified a spectrum of dysplastic changes in the follicular dendritic cells (FDC) of atrophic lymphoid follicles, with some cases showing expansions of FDC networks by CD21 immunostaining. We propose that localized clonal proliferations of stromal elements, particularly follicular dendritic cells, occur in typical HVCD and likely explain the increased incidence of FDC sarcomas in these patients. PMID- 10843294 TI - Renal angiomyolipoma with epithelioid sarcomatous transformation and metastases: demonstration of the same genetic defects in the primary and metastatic lesions. AB - Angiomyolipoma (AML) is a benign neoplasm that occurs either sporadically or in patients with tuberous sclerosis complex (TSC) and shows frequent allelic losses at chromosome arm 16p. It has been suggested recently that the melanogenesis marker-positive perivascular epithelioid cell (PEC) has been found consistently in AML. The authors report a 50-year-old woman without evidence of TSC affected by classic renal AML containing an area composed of atypical epithelioid cells with the same morphoimmunophenotypic characters of PEC. After 7 years from surgical removal of the lesion, the patient developed a local recurrence and successive lung and abdominal metastases that showed morphologic and immunohistochemical features overlapping those of the epithelioid area of the previously removed AML. Genetic analysis showed that the classic AML and its epithelioid area as well as the pulmonary and abdominal metastases shared the same allelic loss on chromosome arm 16p. Based on these findings, the authors view this case as evidence of a malignant transformation of a classic AML with morphologic, immunophenotypic, and genetic demonstration of its clonal origin. PMID- 10843295 TI - Osseous metaplasia in polypoid cholesterosis. PMID- 10843296 TI - Atypical reexcision perineural invasion. PMID- 10843297 TI - Reward discounting as a measure of impulsive behavior in a psychiatric outpatient population. AB - Impulsivity has been operationalized as a choice of an immediate smaller reward over a larger delayed or uncertain reward. This study examined a procedure that measures reward preference under these contingencies in psychiatric outpatients considered either at a high or low risk for engaging in impulsive behavior depending on their psychiatric diagnoses. The participants' rates of delay and uncertainty reward discounting were compared with their performances on a behavioral inhibition task and responses on a self-report personality impulsivity measure. The high-risk participants discounted delayed rewards more sharply and scored higher on the self-report impulsivity measure relative to the low-risk participants. Delay and uncertainty discounting were modestly correlated, but no other relationships were found between the other measures. Results from this study indicate that delay-discounting tasks may be sensitive to at least one form of impulsive behavior. PMID- 10843298 TI - Active cocaine immunization attenuates the discriminative properties of cocaine. AB - Anticocaine antibody, resulting from immunization with the cocaine-keyhole-limpet hemocyanin (KLH) conjugate, weakened the ability of cocaine to act as a discriminative stimulus in rats. Subjects were given extensive training to discriminate 5.0 mg/kg of cocaine from saline prior to immunization. Several weeks following immunization with cocaine-KLH, subjects failed to reliably discriminate cocaine from saline. Nonimmunized control rats retained the ability to discriminate cocaine from saline throughout the experiment. These results further demonstrate that active immunization is effective in blunting cocaine effects. Immunized subjects were able to discriminate 20 mg/kg of cocaine, however, suggesting that anticocaine antibody may be overwhelmed by large cocaine doses. PMID- 10843299 TI - Integrating biological and behavioral factors in alcohol use risk: the role of ALDH2 status and alcohol expectancies in a sample of Asian Americans. AB - Prior studies have shown that the ALDH2*2 genetic variant, most common in individuals of Asian descent, is related to heightened sensitivity to alcohol and can serve as a protective factor against alcohol problems. This study explored the effect of this factor on alcohol expectancies. It was hypothesized that (a) individuals with ALDH2*2 alleles would have lower positive expectancies and higher negative expectancies, (b) expectancies would mediate the ALDH2-drinking relation, and (c) ALDH2 status would moderate the expectancy-drinking relation. Data were collected from 171 Asian American university students. Positive expectancy and ALDH2 status were correlated with alcohol use. Mediation and moderation hypotheses were supported only in the female sample. Results were not significant for negative expectancies. These results indicate that ALDH2 status may protect against drinking by lowering positive expectancies and reducing the expectancy-drinking relationship. PMID- 10843300 TI - Treatment of heroin-dependent poly-drug abusers with contingency management and buprenorphine maintenance. AB - This study targeted poly-drug (cocaine plus heroin) abstinence among buprenorphine-maintained participants with a 12-week voucher-based reinforcement therapy (VBRT) phase versus a yoked control condition. Baseline levels of cocaine and heroin use were significant predictors of treatment outcome, regardless of treatment assignment. Overall, there were no significant group differences on treatment outcome. However, among the subsample that produced one or more poly drug-free urine results, VBRT participants had significantly increased cocaine but not heroin and poly-drug-abstinence, although all results were in the predicted direction. Results suggest that for those who achieve poly-drug abstinence, VBRT may enhance treatment outcome. However, improved interventions, perhaps targeting single-drug abstinence, increasing reinforcement magnitude, or both, may be necessary to promote initial poly-drug abstinence in this population. PMID- 10843301 TI - The role of women's alcohol consumption in evaluation of vulnerability to sexual aggression. AB - The authors examined the impact of alcohol consumption on women's risk perceptions and intended behaviors in a hypothetical situation in which the potential for establishing a relationship with an attractive man was coupled with the potential for sexual aggression. Fifty-nine single women, ages 21-29, were randomly assigned to 1 of 3 beverage conditions: (a) alcohol (dose sufficient to raise blood alcohol level to .08); (b) placebo, in which they were led to believe that they had consumed alcohol but had not; or (c) no alcohol, in which they neither expected nor received alcohol. Compared with women in the no-alcohol condition, women in the alcohol condition (a) rated the male character in the vignette more positively, (b) anticipated less risk and more benefit resulting from a series of behaviors likely to facilitate the relationship while increasing sexual vulnerability (e.g., engaging in consensual sexual activities), and (c) anticipated greater involvement in those behaviors. The placebo appeared to exert similar but weaker effects. PMID- 10843302 TI - Inhibition of smoking-related information in smokers and nonsmokers. AB - Twenty-four smokers and 24 nonsmokers performed a modified version of M. A. Gernsbacher, K. R. Varner, and M. E. Faust's (1990) suppression task, involving presentations of sentences on a computer screen. Each sentence was followed by a word that either was or was not related to the meaning of the sentence. Participants judged whether the word was related to the sentence by pressing either a "yes" or "no" key on a button box. In the experimental sentences, the test word was related to one meaning of the final word of the sentence, but this was not necessarily the meaning intended in the sentence. In half of the experimental sentences, the last word was a smoking-related word (e.g., tar or ashes). Smokers had relatively longer response latencies and lower accuracy scores than nonsmokers when the final word was smoking related, whereas both groups performed similarly on items unrelated to smoking, suggesting that smokers had more difficulty than nonsmokers inhibiting task-irrelevant, smoking-related information but that they did not have a general inability to inhibit irrelevant information. PMID- 10843303 TI - Inhibition conflict and alcohol expectancy as moderators of alcohol's relationship to condom use. AB - Inhibition conflict theory predicts that alcohol will decrease condom use only among individuals who are highly conflicted about using a condom, whereas expectancy theory predicts such an effect only among individuals who hold strong beliefs about alcohol's effects on sexual risk taking. In Study 1, the first of these two theories was tested using a newly developed measure of conflict. Data from 308 college students who reported on the first time they had sexual intercourse with their most recent partner (FMRP) supported the utility of this measure and showed that quantity of alcohol consumed was negatively associated with condom use only among high-conflict individuals. In Study 2, 17- to 25-year old respondents reported on their first sexual intercourse, FMRP, and last intercourse (ns = 465, 1136, and 984, respectively). In a simultaneous test of both inhibition conflict theory and expectancy theory, amount of alcohol consumed was found to be negatively associated with condom use at first intercourse among individuals high in both conflict and expectancy, at FMRP among high-expectancy individuals, and at last intercourse among high-conflict individuals. These results lend partial support to both theories of alcohol's effects and suggest that an integration of these two perspectives will ultimately be required if researchers are to model adequately alcohol's effects on human social behavior. PMID- 10843304 TI - An assessment and comparison of the effects of oxotremorine, D-cycloserine, and bicuculline on delayed matching-to-sample performance in rats. AB - The effects of a muscarinic antagonist (scopolamine), a muscarinic agonist (oxotremorine), an agonist at the N-methyl-D-aspartate receptor site (D cycloserine), and a GABAa antagonist (bicuculline) on working memory were compared using rats performing a delayed matching-to-sample task. When administered on their own, oxotremorine, D-cycloserine, and bicuculline had no effect on performance in the current task. When administered concurrently with scopolamine, oxotremorine (at 1 dose) and bicuculline (at 2 doses) improved accuracy (in terms of percentage correct) by ameliorating the scopolamine-induced increase in response bias. None of the drugs, however, were successful in ameliorating the scopolamine-induced impairment in bias-free recognition performance per se (as measured by Log d). Therefore, none of the drugs examined were able to fully ameliorate all aspects of the memory impairment caused by scopolamine. PMID- 10843305 TI - Relative taste thresholds for ethanol, saccharin, and quinine solutions in three strains of rats nonselected for ethanol: a comparative study. AB - C. P. Richter and K. H. Campbell (1940b) originally defined taste threshold as "the point at which the rats first indicated that they recognized a difference between the distilled water and the solutions" (p. 34). The present study sought to apply this simple behavioral measure to the investigation of strain differences in taste sensitivities, particularly with respect to predictive relationships in ethanol, saccharin, and quinine preference. Fawn-Hooded, Lewis, and Wistar rats were presented with gradual increments in concentration of ethanol (0.01-15%; C. P. Richter & K. H. Campbell, 1940a), saccharin (0.002-3%) or quinine (0.0001-0.0055). Results showed that although intake for saccharin was similar in all strains, consumption of ethanol and quinine differed among the groups. Although previous research has proposed that sweet preference is a promising behavioral marker for ethanol preference, these results suggested that bitter preference may be a more reliable predictor of ethanol preference in rats. PMID- 10843306 TI - "Loss of control" in alcoholism and drug addiction: a neuroscientific interpretation. AB - Considerable neurological evidence indicates that the prefrontal cortex mediates complex "executive" functions including behavioral autonomy and self-control. Given that impairments of self-control are characteristic of alcoholism and other drug addictions, frontal lobe dysfunction may play a significant role in such compulsive behaviors. Consistent with this idea, recent research using brain imaging, neuropsychological testing, and other techniques has revealed that the frontal lobes are particularly vulnerable to the acute and chronic effects of addictive drugs, especially alcohol and cocaine. Evidence implicating a hyperdopaminergic mechanism of acute and chronic drug-induced frontal lobe dysfunction and interactions with premorbid factors and stress are discussed. PMID- 10843307 TI - Improving quality of care for depression in primary care. PMID- 10843308 TI - Are there detectable differences in quality of care or outcome of depression across primary care providers? AB - OBJECTIVE: The objective of this work was to determine whether there are detectable differences among primary care physicians in measures of quality of care or clinical outcome for depressed patients during the first 2 months of treatment with antidepressant medication. METHODS: We studied 1,599 depressed primary care patients initiating antidepressant treatment from 63 family physicians in 4 primary care clinics of a staff-model health maintenance organization. Patients were interviewed 6 to 8 weeks after initiating antidepressant medication with a telephone structured interview that included the Structural Clinical Interview for DSM-IV Diagnoses (SCID). Automated databases of the HMO were used to examine 3 quality-of-care measures: (1) the percentage of patients who had a refill of their antidepressant by 6 weeks, (2) the percentage of patients who had a return visit by 3 weeks, and (3) the percentage of patients having a return visit by 6 weeks. The percentage of patients in each primary care physician panel who had > or =4 persistent DSM-IV depressive symptoms at 6 to 8 weeks was the main clinical outcome variable. To adjust for case-mix differences between physician panels, patient age, gender, and medical comorbidity were controlled for in the analyses. Two covariates were used to adjust for differences in patients' clinical severity: self-report of > or =2 prior depressive episodes and an SCID diagnosis of major depression during the patient's worst episode in the last 2 years. Physician age, gender, and part-time versus full-time practice were also used as covariates. RESULTS: The wide observed range of variability in quality-of-care and clinical outcome measures by physician practice decreased markedly in the statistical model that controlled for patient-level covariates and differences in the number of patients seen per provider. We did not detect differences in physician practice for the percentage of patients who had a return visit by 3 or 6 weeks, the percentage of patients who had a refill of their antidepressant prescription by 6 weeks, or the percentage of patients with an adverse clinical outcome of depression. CONCLUSIONS: We did not find important differences in measures of quality of care or patient outcomes by physician. These results may have implications for the use of physician profiling and other forms of physician report cards. PMID- 10843309 TI - Cost-outcome analysis in injury prevention and control: eighty-four recent estimates for the United States. AB - OBJECTIVES: The objectives of this study were to review cost-outcome analyses in injury prevention and control and estimate associated benefit-cost ratios and cost per quality-adjusted life-year. DATA SOURCES: Medline and Internet search, bibliographic review, and federal agency contacts identified published and unpublished studies from 1987 to 1998 for the United States. Studies of low quality and analyses of occupational, air, rail, and water transport safety programs were excluded. MEASURES: Selected results were recomputed to increase discount rate, benefit category, and benefit estimate comparability and to update injury incidence rates. RESULTS: More than half of the 84 injury prevention measures reviewed yielded net societal cost savings. Twelve measures had costs that exceeded benefits. Of 33 road safety measures analyzed, 19 yielded net cost savings. Of 34 violence prevention approaches studied, 19 yielded net cost savings, whereas 8 had costs that exceeded benefits. Interventions with the highest benefit-cost ratios included juvenile delinquent therapy programs, fire safe cigarettes, federal road and traffic safety program funding, lane markers painted on roads, post-mounted reflectors on hazardous curves, safety belts in front seats, safety belt laws with primary enforcement, child safety seats, child bicycle helmets, enforcement of laws against serving alcohol to the intoxicated, substance abuse treatment, brief medical interventions with heavy drinkers, and a comprehensive safe communities program in a low-income neighborhood. Studies of cost-saving measures do not exist for several injury types. CONCLUSIONS: Injury prevention often can reduce medical costs and save lives. Wider implementation of proven measures is warranted. PMID- 10843310 TI - One thousand health-related quality-of-life estimates. AB - OBJECTIVE: Analysts performing cost-effectiveness analyses often do not have the resources to gather original quality-of-life (QOL) weights. Furthermore, variability in QOL for the same health state hampers the comparability of cost effectiveness analyses. For these reasons, opinion leaders such as the Panel on Cost-Effectiveness in Health and Medicine have called for a national repository of QOL weights. Some authors have responded to the call by performing large primary studies of QOL. We take a different approach, amassing existing data with the hope that it will be combined responsibly in meta-analytic fashion. Toward the goal of developing a national repository of QOL weights to aid cost effectiveness analysts, 1,000 health-related QOL estimates were gathered from publicly available source documents. METHODS: To identify documents, we searched databases and reviewed the bibliographies of articles, books, and government reports. From each document, we extracted information on the health state, QOL weight, assessment method, respondents, and upper and lower bounds of the QOL scale. Detailed guidelines were followed to ensure consistency in data extraction. RESULTS: We identified 154 documents yielding 1,000 original QOL weights. There was considerable variation in the weights assessed by different authors for the same health state. Methods also varied: 51% of authors used direct elicitation (standard gamble, time tradeoff, or rating scale), 32% estimated QOL based on their own expertise or that of others, and 17% used health status instruments. CONCLUSIONS: This comprehensive review of QOL data should lead to more consistent use of QOL weights and thus more comparable cost effectiveness analyses. PMID- 10843311 TI - Demographic predictors of eye care utilization among women. AB - BACKGROUND: Visual impairment may be preventable or treatable with timely intervention. Differences in the use of eye care services may play a role in fostering the racial and socioeconomic gap in the burden of visual impairment in the United States. OBJECTIVE: The objective of this study was to examine the frequency of eye examinations in women and how this varies with age, race, marital status, geographic region, profession, education, and income. SUBJECTS AND METHODS: We used logistic regression models to examine data obtained by mailed questionnaires from 39,876 female health professionals participating in the Women's Health Study. RESULTS: Most women (83%) had an eye examination within the past 2 years. The likelihood of having an eye examination in the past 2 years increased with age (odds ratio [OR] = 2.59 for age > or =75 years versus those <50 years; P [trend] <0.0001), higher education (OR = 1.27 for master's degree versus licensed nurse training; P [trend] = 0.0004), and higher household income (OR = 1.85 for > or =$100,000 versus <$10,000; P [trend] <0.0001). Women from the south were less likely to have had an eye examination than those from the west (OR = 0.92; P = 0.03). Compared with whites, Asian/Pacific Islanders were less likely (OR = 0.76; P = 0.02) and blacks more likely (OR = 1.27; P = 0.02) to have had an eye examination within 2 years. CONCLUSIONS: Age, education, income, race/ethnicity, and region of residence were independent predictors of having had an eye examination in the past 2 years. Known medical and ocular problems appeared to explain the association with age but not the other findings, although the clinical significance of these associations was not determined in the present study. Additional research on determinants of eye care-seeking behavior could help in devising new strategies to encourage preventive behaviors, especially among groups at higher risk of visual impairment. PMID- 10843312 TI - Does diagnostic information contribute to predicting functional decline in long term care? AB - BACKGROUND: Compared with the acute-care setting, use of risk-adjusted outcomes in long-term care is relatively new. With the recent development of administrative databases in long-term care, such uses are likely to increase. OBJECTIVES: The objective of this study was to determine the contribution of ICD 9-CM diagnosis codes from administrative data in predicting functional decline in long-term care. RESEARCH DESIGN: We used a retrospective sample of 15,693 long term care residents in VA facilities in 1996. METHODS: We defined functional decline as an increase of > or =2 in the activities of daily living (ADL) summary score from baseline to semiannual assessment. A base regression model was compared to a full model enhanced with ICD-9-CM codes. We calculated validated measures of model performance in an independent cohort. RESULTS: The full model fit the data significantly better than the base model as indicated by the likelihood ratio test (chi2 = 179, df = 11, P <0.001). The full model predicted decline more accurately than the base model (R2 = 0.06 and 0.05, respectively) and discriminated better (c statistics were 0.70 and 0.68). Observed and predicted risks of decline were similar within deciles between the 2 models, suggesting good calibration. Validated R2 statistics were 0.05 and 0.04 for the full and base models; validated c statistics were 0.68 and 0.66. CONCLUSIONS: Adding specific diagnostic variables to administrative data modestly improves the prediction of functional decline in long-term care residents. Diagnostic information from administrative databases may present a cost-effective alternative to chart abstraction in providing the data necessary for accurate risk adjustment. PMID- 10843313 TI - Nursing home-to-nursing home transfers: prevalence, time pattern, and resident correlates. AB - OBJECTIVES: To characterize the time pattern of nursing home-to-nursing home transfers and assess which resident characteristics are associated with transfers. METHODS: Minimum Data Set assessments of all Maine and New York nursing home residents were obtained for 1994-1996. The hazard rate for nursing home transfers was estimated by nonparametric statistical techniques, censored at loss to follow-up. Comparisons of resident characteristics were made between those who transferred and those who stayed at their initial facility. RESULTS: Residents of Maine nursing homes were considerably more likely to transfer than were New York residents. Transfer rates declined during the first 2 years after admission and remained stable thereafter. Correlates of transfers were similar across states. Residents who transferred were more likely to be male, to be married, to be younger, to have better cognitive and physical health, to have Medicare or private payment sources (vs. Medicaid), and to have pressure ulcers. Rural location did not affect the likelihood of transfer. CONCLUSIONS: This study provides the most detailed information to date on the prevalence, timing, and correlates of nursing home transfers. These transitions occur most frequently early in the stay but continue at a lower rate even among long stayers. This information is useful for understanding lifetime dynamics of long-term-care utilization. Several barriers to mobility appear to be present (eg, less generous payment source, health limitations, and absence of a spouse). The higher transfer rates observed in Maine might imply that institutional or other factors limit the mobility of New York residents. PMID- 10843314 TI - Sociodemographic factors and the assignment of do-not-resuscitate orders in patients with acute myocardial infarctions. AB - OBJECTIVES: This study examined the impact of sociodemographic and clinical factors, measured at the individual or ecological (zip code) level, on the assignment of do-not-resuscitate (DNR) orders. DESIGN: This was a retrospective study (analysis of secondary data). SUBJECTS: We used a probability sample of 974 patients admitted to 30 medium to large California hospitals with acute myocardial infarctions in 1990 to 1991; the sample was originally designed to validate risk adjustment with administrative data. METHODS: Multivariate logistic regression was used to adjust DNR assignment for age, gender, race, probability of death, functional impairments, payment source, hospital teaching status, and ecological measures of educational attainment, home ownership, and income. RESULTS: DNR assignment was inversely associated with black race and positively associated with age, probability of death, cognitive impairment, and poor nutritional status. When the probability of death was very low, DNR orders were assigned less frequently to men than to women (odds ratio [OR], 0.4; 95% confidence interval [CI], 0.2 to 0.7 at probability of death = 0.10). However, men were significantly more likely to receive a DNR order than women when the probability of death was very high (OR, 4.4; 95% CI, 1.2 to 16.3 at probability of death = 0.90). CONCLUSIONS: Older, white, sicker, or functionally impaired patients receive DNR orders more often than younger, black, healthier, or functionally intact patients do. Adjusting for these factors, DNR assignment is associated with gender through an interaction involving the probability of death. Future studies should reexamine the impact of these factors on DNR assignment and explore the role of patient values and patient-physician communication barriers. PMID- 10843315 TI - Effect of future costs on cost-effectiveness of medical interventions among young adults: the example of intensive therapy for type 1 diabetes mellitus. AB - OBJECTIVES: Recent research based on a lifetime utility maximization model has suggested that cost-effectiveness analyses should account for all future costs, including medical costs for related and unrelated illnesses and nonmedical costs. This work has also shown that analyses that omit future costs are biased to favor interventions among the elderly that extend life over interventions that improve quality of life. However, the effect of including future costs on the cost effectiveness of interventions among the young has not been studied. This article examines the effect of including future costs on the cost-effectiveness of intensive therapy for type 1 diabetes mellitus among young adults. METHODS: By modifying a cost-effectiveness model based on the Diabetes Control and Complications Trial to include future costs, the effect of including future costs on the cost-effectiveness of intensive therapy for type 1 diabetes mellitus among young adults was examined. Future costs added to the model included future costs for medical expenditures for illnesses unrelated to diabetes and future nonmedical expenditures net of earnings. RESULTS: Intensive therapy among young adults led to approximately equal increases in the expected number of years lived before age 65, when people generally produce more than they consume, and after age 65, when the opposite tends to hold. Because the discounted value of savings due to lower mortality before age 65 exceeded the discounted value of later increases in costs due to lower mortality after age 65, accounting for future costs decreased the cost-effectiveness ratio from $22,576 to $9,626 per quality adjusted life-year. CONCLUSIONS: The inclusion of future costs can significantly improve the cost-effectiveness of interventions that decrease mortality among young adults. The common practice of excluding future costs may bias cost effectiveness analyses against such interventions. PMID- 10843316 TI - Texas physician immunization practices. AB - INTRODUCTION: Immunization levels among young children could be improved if physicians administered immunizations at both well and urgent visits, provided simultaneous vaccinations, and knew the guidelines on valid contraindications, part of the Standards for Pediatric Immunization Practices (Standards). We report on a survey measuring the immunization knowledge and practice of Texas pediatricians, family practitioners, and general practitioners. METHODS: A survey was mailed to a random sample of physicians. Questionnaire items covered 8 of the 18 Standards. RESULTS: The response rate was 62% (608 of 976 eligible physicians). The mean summary adherence-to-Standards score was 4.1 for pediatricians, 3.6 for family practitioners, and 3.0 for general practitioners. Specialty and practice location were significant predictors of the summary score, whereas gender, managed-care participation, and graduation year were not. DISCUSSION: Our study results suggest that in Texas, pediatricians and rural physicians most often adhere to the Standards. Improving the reimbursement level of administrative costs for immunization delivery, educating physicians on the Standards, and encouraging the use of patient immunization tracking systems are actions that could potentially improve immunization rates. PMID- 10843317 TI - A check-list of the nematode parasites of South African Serpentes (snakes) and Sauria (lizards). AB - Published records, in combination with own data have been brought together to provide data on parasite/host relationships of reptiles that occur in the Republic of South Africa. A total of 62 nematode species belonging to 23 genera and 11 families are recorded from 20 snake and 21 lizard species. The genera Kalicephalus, Spauligodon, Ophidascaris and Abbreviata are especially well represented with between five and eight species per genus. The most nematode species were recorded from the flap-neck chameleon, Chamaeleo dilepis (eight), the puff-adder, Bitis arietans (eight) and the water monitor, Varanus niloticus (seven). All synonyms of parasites and hosts are given. PMID- 10843318 TI - A newly developed odour-baited "H trap" for the live collection of Glossina brevipalpis and Glossina austeni (Diptera: Glossinidae) in South Africa. AB - A new trap, named the "H trap", was developed at Hellsgate Tsetse Research Station in South Africa for the simultaneous collection of live Glossina brevipalpis Newstead and Glossina austeni Newstead. Its design followed an evaluation of the responses of the two species towards traps that are used elsewhere in Africa for the collection of other tsetse fly species. These traps were found at Hellsgate to be unsuitable for capturing both G. brevipalpis and G. austeni. Some new trap designs and many modifications of these were tested, most of which were unsuccessful. The odour-baited blue and black H trap represents a different approach for trapping tsetse flies as it is fitted with lateral cones of white netting which induce the flies to take a more horizontal flight path once they have entered the trap, instead of the vertical flight paths they assume in existing tsetse fly traps. A number of modifications of the prototype H trap were devised (H1-H5), before the final design was established. Catches of up to 76 G. brevipalpis and 37 G. austeni were obtained per trap on a single day with the H3 modification. Further modifications improved on the trap's efficiency to capture G. brevipalpis and G. austeni. The final modification caught a record number of 180 G. brevipalpis and 57 G. austeni on a single day. PMID- 10843319 TI - Investigation of biological samples for monofluoroacetate and Dichapetalum cymosum poisoning in southern Africa. AB - A need has existed for the accurate identification of monofluoroacetate (MFA) poisoning in southern Africa. The development of a new method for the determination of MFA has made the analysis of a variety of biological samples (n = 50) feasible. The method has been used in the laboratory over 24 months. Monofluoroacetate was present in 66% of samples from cases of suspected poisoning, reflecting the extent of the problem. Stability of MFA in samples was also determined so as to have a time-bound baseline for the acceptance of samples submitted. It was found that there was a decrease in the level of MFA and, after 14 days at room temperature, only 50% of the spiked dose could be identified. It is suggested that samples be analyzed within 7 days of mortality if they not kept frozen. PMID- 10843320 TI - Parasites of South African wildlife. XVI. Helminths of some antelope species from the Eastern and Western Cape Provinces. AB - The numbers and species of helminths recovered from one black wildebeest, Connochaetes gnou, three eland, Taurotragus oryx, 18 mountain reedbuck, Redunca fulvorufula, one red hartebeest, AIcelaphus buselaphus and two springbok, Antidorcas marsupialis, in the Mountain Zebra National Park, Eastern Cape Province; two black wildebeest, two grey rhebuck, Pelea capreolus, two mountain reedbuck and four springbok in the Karoo National Park, Western Cape Province; two bontebok, Damaliscus pygargus dorcas, two eland, two gemsbok, Oryx gazella and two springbok in the West Coast National Park, Western Cape Province; and a single springbok on a farm near Bredasdorp, Western Cape Province, are recorded. Nematodes belonging to a total of 12 genera and 20 species were identified. A single cestode was also recovered. Sixteen new host associations are recorded for the nematodes and one for the cestode Moniezia benedeni. Nematodirus spathiger had the widest host spectrum and with the exception of black wildebeest, was collected from all the host species examined. PMID- 10843321 TI - Suspected sheep-associated malignant catarrhal fever in a zero-grazed dairy herd in Kenya. AB - An outbreak of a disease characterised by very high mortality occurred in a group of nine calves (1B4 months old) in a zero-grazing unit 2-3 weeks after an introduction of an apparently healthy alien sheep into the calf pen. Five of the six calves which contracted the disease died. The main clinical signs observed were marked depression, persistently high body temperature (40,5-41,5 degrees C), copious mucopurulent nasal and ocular discharges, dyspnoea, bilateral keratoconjunctivitis with corneal opacity, enlargement of the superficial lymph nodes and marked erythema and/or superficial erosions of the buccal mucosae. At necropsy there were lesions in the upper respiratory and digestive tracts, lymph nodes, brain, eyes, liver, kidneys and the urinary bladder. The lesions were histopathologically characterized by fibrinoid vasculitis which was accompanied by lymphocytic infiltration in the parenchyma of the affected tissues. Based on the evidence of contact between the calves and the recently introduced foreign sheep, the characteristic clinical signs and histopathological findings, a diagnosis of sheep-associated malignant catarrhal fever was made. PMID- 10843322 TI - Oribatid mites (Acari, Oribatida) as intermediate hosts of tapeworms of the Family Anoplocephalidae (Cestoda) and the transmission of Moniezia expansa cysticercoids in South Africa. AB - Six species of adult oribatid mites (Galumna racilis, Kilimabates pilosus, Kilimabates sp., Scheloribates fusifer, Muliercula ngoyensis and Zygoribatula undulata) and two immature stages belonging to the superfamilies Galumnoidea and Ceratozetoidea were isolated from a lawn (mixed Pennisetum and Cynodon spp.) at Onderstepoort Veterinary Institute, South Africa. The mites were subsequently used in an infection trial using Moniezia expansa eggs. Tapeworm cysticercoids were recovered in G. racilis, K. pilosus, Kilimabates sp., S. fusifer, M. ngoyensis and Z. undulata, as well as in immatures of Ceratozetoidea. The percentage of infected mites was 7,6, 6,3, 16,4, 66,7, 57,1, 60,0 and 46,7% respectively. Immatures of Galumnoidea did not become infected. The highest number of cysticercoids isolated from one individual was six from an adult S. fusifer. PMID- 10843323 TI - Leishmania major infections in Phlebotomus duboscqi fed on murine models immunized with L. major subcellular antigens and sandfly gut antigens. AB - The ability of antibodies in bloodmeals of mice and hamsters immunized with Leishmania major subcellular fractions and sandfly (Phlebotomus duboscqi) gut antigens to inhibit development of L. major in its vector P. duboscqi was examined. Antibodies from animals immunized with either L. major subcellular fractions alone or sandfly gut antigen alone were not very effective in inhibiting development of L. major in the sandfly. When P. duboscqi were fed on blood from animals immunized with both parasite flagella and sandfly gut antigen, development of L. major was significantly inhibited (P<0,05). Control sandflies fed on naive animals displayed a normal pattern of parasite development to the metacyclic stage. Electron microscopy studies showed that one of the mechanisms through which antisandfly gut antibody can cause inhibition of parasite development is by lysing sandfly gut epithelium. This study has demonstrated that it is possible to reduce transmission of leishmaniosis through immunization against both the parasite and its sandfly vector. PMID- 10843324 TI - The prevalence of different African horsesickness virus serotypes in the Onderstepoort area near Pretoria, during an outbreak of African horsesickness in South Africa in 1995/1996. AB - During 1995/1996 parts of South Africa experienced exceptionally high rainfall. Large numbers of Culicoides midges were seen and an outbreak of African horsesickness (AHS) followed. In the Onderstepoort area, near Pretoria in Gauteng, a number of horses died of suspected AHS. Virus isolation and typing was done from blood and/or organ samples of 21 suspected cases as well as from five zebra which were kept in the area. Virus was isolated from 14 of the 21 suspected cases but not from the zebra. The neutralizing antibody response of the zebra to the nine different African horsesickness virus (AHSV) serotypes was determined. Results indicated the highest prevalence of serotypes 2 and 4 followed by serotypes 1, 6 and 9. Reverse transcription polymerase chain reaction (RT-PCR) was performed on total RNA extracted from blood samples of the zebra. AHSV RNA was indicated in three of five zebra by agarose gel electrophoresis analysis of amplicons and in four of five zebra after Southern blot hybridization using a 32P labelled probe. RT-PCR can be used together with serological techniques in studies of AHS to further clarify the epizootiology of the disease. PMID- 10843325 TI - Suggested dosage rates of melarsoprol in the treatment of mice experimentally infected with Trypanosoma brucei gambiense. AB - One group of BALB/c mice infected with a highly virulent strain of Trypanosoma brucei gambiense were treated intraperitoneally with three series of three injections (each injection of 10 mg/kg) of Mel-B separated by seven days of rest, while a second group was treated once by a single injection All the Mel-B treated mice in both experiments were negative for parasites when examined using either the wet blood film or buffy coat methods, but were intermittently PCR positive during the sampling period. We encourage the use of a repeat negative PCR test over a one month period in combination with corroborative clinical and parasitological investigation to be suggestive of cure in experimental animals previously infected with trypanosomosis. In view of the exorbitant costs of Mel-B and its extreme toxicity, we recommend that Mel-B be given as one course of two injections (each equivalent to 10 mg/kg) separated by 2 d of rest in experimentally infected rodent models. PMID- 10843326 TI - Helminth parasites of domestic pigeons (Columba livia domestica) in Sebele, Gaborone, Botswana. AB - Twelve adult domestic pigeons from Sebele, Gaborone, Botswana, were examined for the presence of helminth parasites. The cestode genus Raillietina and two species of nematodes, Dispharynx spiralis and Ascaridia columbae were recovered. Most pigeons (75 %) were infected with Raillietina spp. often in concurrence with A. columbae. PMID- 10843327 TI - Evaluation of the attenuating properties of selected Greek clays for toxic inorganic elements in landfill sites. AB - Heavy metal attenuation properties of selected clay material collected from miscellaneous Greek sites is investigated and tested in the laboratory for their suitability, either as liners in hydrologically unsafe sites or as earth covers for sanitary landfill sites. Eleven potentially hazardous elements (As, Be, Cd, Cr, Cu, Hg, Ni, Pb, Se, V, and Zn) generated by a co-disposal landfill leachate have been considered. Experimental column and static equilibrium methods for the determination of dispersion and adsorption are described. Molecular diffusion dominates the migration phenomena with a velocity range between 1.3 x 10(-5) and 3.5 x 10(-4) cm/s throughout the experiments. A simple way to evaluate dispersion coefficients from breakthrough curves gave values of between 3.90 x 10(-6) and 3.5 x 10(-4) cm2/s, with a mean value of 1.5 x 10(-5). Static adsorption equilibrium studies supported by column runs showed that Freundlich (F = kCn) isotherms express in a better way the assimilative capacities of the tested clays, with k and n values ranging from 0.06 to 1.99 and 0.55 to 1.48 correspondingly. Mathematical models involving non-linear parabolic equations are involved. The experimental data, together with finite difference techniques and some physical clay characteristics, produced trilinear textural diagrams and predictive flow transport convection-dispersion breakthrough curves for a quick estimation of the attenuating properties of clays for heavy metals. PMID- 10843328 TI - Sediment-air equilibrium partitioning of semi-volatile hydrophobic organic compounds. Part 1. Method development and water vapor sorption isotherm. AB - Contaminated sediments that become exposed to air as a result of dredging and disposal in confined disposal facilities are potential sources of air pollution. A critical parameter to develop emission estimation models is the equilibrium partition coefficient of contaminants, between sediment and air. In this first of two articles, we present a method, based on gas saturation in a flowing stream, to study both the adsorption of water and semi-volatile organic compounds on a sediment from the Campus Lake, Baton Rouge, LA, USA. The experimental set-up was used to determine the adsorption isotherm for water partitioning between sediment and pore-air. A detailed characterization of the sediment surface area and pore volume was used to develop an adsorption-condensation model for predicting water sorption on sediment. The model was used to estimate the importance of water adsorption on mineral surfaces and condensation in pores. This information serves, in the accompanying second article in the series, as the basis for the modeling of the partitioning of phenanthrene, and dibenzofuran. PMID- 10843329 TI - Sediment-air equilibrium partitioning of semi-volatile hydrophobic organic compounds part 2. Saturated vapor pressures, and the effects of sediment moisture content and temperature on the partitioning of polyaromatic hydrocarbons. AB - A gas saturation methodology was used to determine the sediment/air partition coefficient (K(SA)) for phenanthrene and dibenzofuran on a local sediment from the Campus Lake, Baton Rouge. The effects of sediment moisture content, air relative humidity and temperature on K(SA) for phenanthrene were ascertained. The saturated vapor pressures of the compounds were also measured. The sediment moisture content had a striking effect on K(SA); increasing sediment moisture content decreased K(SA). Temperature also had a dramatic effect on K(SA). The variation with temperature was used to evaluate the heats of adsorption on both 'dry' (< 0.8% by mass of water) and 'wet' (> 6% by mass of water) sediments. The heat of adsorption for phenanthrene normalized to unit molecular surface area indicated for the physically adsorbed organic compound was seven times smaller than that for a water molecule. The experimental value of K(SA) was used to test a proposed theoretical model. A good agreement was observed for both phenanthrene and dibenzofuran. The model can be extended to other compounds and sediment types for prediction of air emissions from exposed sediment and dredged materials. PMID- 10843330 TI - The distribution of mercury and other trace metals in the sediments of the Mersey Estuary over 25 years 1974-1998. AB - Mercury and other heavy metals have been monitored in the surface sediments of the Mersey Estuary in NW England for a period of 25 years, using a consistent methodology and sampling grid. This has produced one of the most comprehensive data sets available in the literature and demonstrated some of the difficulties associated with undertaking long-term environmental monitoring. The data indicate that the concentration of mercury and other metals in the sediments of this estuary are strongly correlated to the organic matter and particle size content. This has resulted in the metal distribution patterns reflecting the sediment characteristics and dynamics, irrespective of the position of the input sources. The trend data has indicated a continual, although somewhat uneven, decline in the concentration of most of the metals studied. Remobilisation of previously consolidated saltmarsh sediments can cause significant perturbations in contaminant reduction trends, and needs to be considered in the design of long term monitoring programmes. PMID- 10843331 TI - Sorption of phenols to dissolved organic matter investigated by solid phase microextraction. AB - The sorption of phenol and different halogenated phenols to natural organic matter of a brown water lake (HO14), of a compost extract, of Aldrich humic acid (Aldrich-HA), and to the protein bovine serum albumin (BSA) was investigated using solid phase microextraction (SPME). The limit of determination for the SPME analysis was < 15 microg/l for all phenols investigated. The extraction coefficients K(F) were calculated according to a first-order extraction kinetics. In general, the extraction equilibrium was established faster due to the presence of dissolved organic matter (DOM). The highest sorption capacity of phenols was observed for BSA with log K(OC) values in the range between 2 and 6. For the compost extract and HO14 only a small sorption of the investigated phenols was determined. On the other hand, Aldrich humic acid showed a reasonable sorption of phenols with log K(OC) values between 2 and 3. The sorption to DOM decreased when the pH of the solution was increased. PMID- 10843332 TI - Multielement analysis in cereals and pulses by k0 instrumental neutron activation analysis. AB - The concentrations of some elements in a few varieties of cereals and pulses are determined by Instrumental Neutron Activation Analysis using a single comparator method (k0-standardised NAA method). A total of 15 elements are measured. The method was validated by analysing the Standard Reference Material (SRM-1571) of NIST; the results are within +/-10% of the reported values for the majority of the elements. The measured concentrations of major and minor elements are analysed in terms of the average intake of mineral content and the role of these elements in terms of the nutritional value. PMID- 10843333 TI - The distribution of micro-organic contaminants in river bed-sediment cores. AB - There is little information available on the distributions of micro-organic contaminants in river bed-sediments below a depth of 5 cm. The aim of this study is to determine the concentrations and distributions of contaminants in river bed sediments up to 1 m depth and to make an assessment of the results. The approach taken was to collect five cores from each of two lowland rivers in southern England, the first a rural river and the second a dominantly urban catchment. The cores were analysed for micro-organic compounds and sediment/sediment porewater characteristics. Compounds detected were polyaromatic hydrocarbons (naphthalene, fluoranthene and pyrene), pesticides (carbaryl, linuron, fenpropimorph, the synthetic pyrethroids, and prometryn), and non-ionic surfactant residue (nonylphenol). A particularly important finding was that some micro-organic contaminants penetrated to depths of 1 m, and in one of the rivers they were detected in undisturbed Eocene substrata. The more hydrophobic contaminants showed a clear depth distribution with higher concentrations towards the top of the cores. The less hydrophobic contaminants demonstrated no systematic trend suggesting they had become soluble in porewaters and subsequently travelled within the cores. Partition between the porewater and sediment appeared to be controlling the distribution of the compounds with depth. PMID- 10843334 TI - Acidic and alkaline precipitation in the Cilician Basin, north-eastern Mediterranean Sea. AB - Samples from precipitation events collected at Erdemli during February 1996-June 1997 were analyzed to determine their particulate aluminium content, in addition to pH and conductivity measurements. Backward air mass trajectories corresponding to the rainy days were analyzed to determine potential source regions of acidic and alkaline constituents transported to the Cilician Basin. Approximately 28% of the rain samples were found to be acidic and the trajectories associated with half of the acid precipitation events were from the Mediterranean Basin and the Balkan Peninsula, while the other half were from the Anatolian mainland and local sources. Rain samples were found to be alkaline (58%), with their trajectories originating from North Africa and the Middle East. As a result of its CaCO3 content, mineral dust from these arid regions significantly increased the pH of rainwater. PMID- 10843335 TI - Characteristics of indoor and outdoor airborne fungi at suburban and urban homes in two seasons. AB - Literature has suggested association between damp environments, microbial exposure, and higher prevalence of respiratory symptoms and diseases. The study began by evaluating the airborne fungal concentrations at urban and suburban areas of a typical metropolitan city in southern Taiwan for the estimation of related health risks. A group of representative homes, based on the housing characteristics questionnaires completed earlier, were selected from two parts of the city; urban and suburban. Burkard sampler (BURKARD, Rickmansworth, England) was used to collect airborne fungi onto agar plates with malt-extract. After incubation and identification, concentrations of airborne fungi were calculated as CFU/m3. The geometric mean (GM) concentration for indoors was 8946 (4372 18,306) CFU/m3 in winter and 4381 (1605-11,956) in summer. For outdoors, it was 11,464 (5767-22,788) CFU/m3 in winter and 4689 (1895-11,603) in summer. In summer, the total fungal concentration, both indoors and outdoors of suburban homes, were significantly higher than those of urban homes. The dominant fungi contributing to such a difference were indoor Cladosporium spp. and outdoor Penicillium spp. (P < 0.01). The indoor/outdoor ratio (I/O) was similar in two areas except for Penicillium spp. in winter and Aspergillus spp. in summer; both higher in the suburban area. Significantly higher levels of airborne fungi were observed in this region than those seen in northern Taiwan or other parts of the world. Future investigations are needed to further examine the effects of these exposures on the related health problems. PMID- 10843336 TI - Limited seasonality effects on blood lead for a small cohort of female adults and children. AB - Many blood lead surveys, especially during the 1970s and 1980s have shown variations of up to 35% in blood lead concentration, with higher values in summer over winter. We have monitored 13 adult females and seven children for periods from 348 to 1337 days as non-pregnant controls in a longitudinal study of mobilization of lead from the maternal skeletal during pregnancy and lactation. Samples of blood, 6-day duplicate diet and environmental samples were analyzed by high-precision thermal ionization mass spectrometry for lead isotope ratios and lead concentrations. There was no statistically significant difference between seasons for blood lead concentrations and dietary intake although there were small differences in the isotopic composition for blood. One explanation for the lack of a seasonal effect in blood lead of our cohort may be the absence of climatic extremes in Sydney. The minimal effects from seasonality observed in this cohort make this an especially useful cohort within which to study effects that could be obscured by seasonal factors. PMID- 10843337 TI - Selection and evaluation of air pollution exposure indicators based on geographic areas. AB - Geographic exposure indicators (GEIs) use point estimates of ambient air pollutant concentrations to characterize the exposure of populations residing within a specified area. Both zone- and proximity-type GEIs have been widely employed in epidemiological studies and other applications to identify regions or populations at high risk. Their use requires a number of assumptions, for example, pollutant concentrations should be homogeneous within the area, and concentrations should differ between areas in a predictable manner. These assumptions have not been rigorously examined. This paper evaluates the most common types of GEIs as surrogate measures of ambient air pollutant exposures. Statistical measures proposed to evaluate GEIs include accuracy, homogeneity, misclassification and statistical power. GEIs and statistical measures are evaluated in two case studies that use different air pollution sources and an air quality dispersion model. The case studies show that pollutant levels may vary substantially within a small area, and significant errors and exposure misclassification may result if the GEI represents a large geographic area. GEIs based on residential proximity to a pollution source should not be used for elevated emission sources, and the use of proximity measures is discouraged for ground level sources. A systematic evaluation is suggested to evaluate and improve the accuracy of the GEIs used in epidemiological and other applications. PMID- 10843338 TI - Industrial fluorosis in cattle and buffalo around Udaipur, India. AB - Signs of dental discolouration, difficulty in mastication, bony lesions, lameness, debility and mortality in domesticated animals, reared around superphosphate fertiliser plants located approximately 15 km north of Udaipur, Rajasthan prompted us to investigate for the occurrence of fluorosis. Out of 166 animals clinically examined, the prevalence rate was 17.4% (4/23) in calves below 1 year of age, 37.2% (16/43) in cattle between 1 and 3 years, 61.3% (46/75) in cattle above 3 years and 72% (18/25) in buffalo above 1 year. Dental fluorosis was common in buffalo compared to cattle of all the age groups. Fluoride levels in fodder and water, consumed by the animals were much higher than the recommended permissible limit. Mean fluoride concentrations in serum and urine were 1.53 +/- 1.27 and 26.4 +/- 6.17 mg l(-1) in calves below 1 year of age, 0.56 +/- 0.17 and 26.2 +/- 3.86 mg l(-1) in cattle of 1-3 years, 0.49 +/- 1.13 and 27.5 +/- 4.63 mg l(-1) in cattle above 3 years and 0.60 +/- 0.07 and 28.6 +/- 4.73 mg l(-1) in buffalo over 1 year, respectively. The values were significantly (P < 0.01) higher than those of control animals kept over a 15-km distance from the factories. Fluoride concentrations in the environmental sample collected from the affected locality were 534.4 +/- 74.9 mg kg(-1) in fodder, 1.19 +/- 0.29 mg l(-1) in pond water and 0.479 +/- 0.351 mg l(-1) in tube well water. It was concluded that the consumption of fodder and water contaminated by the fumes and dusts emitting from superphosphate fertiliser plants resulted in the development of chronic fluorotic lesions in cattle and buffalo. PMID- 10843339 TI - A regression-based method for mapping traffic-related air pollution: application and testing in four contrasting urban environments. AB - Accurate, high-resolution maps of traffic-related air pollution are needed both as a basis for assessing exposures as part of epidemiological studies, and to inform urban air-quality policy and traffic management. This paper assesses the use of a GIS-based, regression mapping technique to model spatial patterns of traffic-related air pollution. The model--developed using data from 80 passive sampler sites in Huddersfield, as part of the SAVIAH (Small Area Variations in Air Quality and Health) project--uses data on traffic flows and land cover in the 300-m buffer zone around each site, and altitude of the site, as predictors of NO2 concentrations. It was tested here by application in four urban areas in the UK: Huddersfield (for the year following that used for initial model development), Sheffield, Northampton, and part of London. In each case, a GIS was built in ArcInfo, integrating relevant data on road traffic, urban land use and topography. Monitoring of NO2 was undertaken using replicate passive samplers (in London, data were obtained from surveys carried out as part of the London network). In Huddersfield, Sheffield and Northampton, the model was first calibrated by comparing modelled results with monitored NO2 concentrations at 10 randomly selected sites; the calibrated model was then validated against data from a further 10-28 sites. In London, where data for only 11 sites were available, validation was not undertaken. Results showed that the model performed well in all cases. After local calibration, the model gave estimates of mean annual NO2 concentrations within a factor of 1.5 of the actual mean (approx. 70 90%) of the time and within a factor of 2 between 70 and 100% of the time. r2 values between modelled and observed concentrations are in the range of 0.58 0.76. These results are comparable to those achieved by more sophisticated dispersion models. The model also has several advantages over dispersion modelling. It is able, for example, to provide high-resolution maps across a whole urban area without the need to interpolate between receptor points. It also offers substantially reduced costs and processing times compared to formal dispersion modelling. It is concluded that the model might thus be used as a means of mapping long-term air pollution concentrations either in support of local authority air-quality management strategies, or in epidemiological studies. PMID- 10843340 TI - An evaluation of critical loads of soil acidity in areas of high sea salt deposition. AB - The empirical and mass balance approaches to setting critical loads of acidity for mineral soils have been evaluated using field data from forest sites in Wales. Using the Simple Mass Balance Equation (SMBE) with Sitka spruce as the biological target, critical loads ranged between 2.3 and 9.8 keq H+ ha(-1) year( 1) compared to mapped empirical critical loads which ranged between 0.2 and 0.5 keq H+ ha(-1) year(-1). At all sites the empirical critical load was exceeded with respect to deposited sulfur acidity. There were no exceeded sites for the SMBE critical loads. The big differences between the two methods arise from the large ANC leaching term in the SMBE model which is determined by the relatively low (Ca + Mg + K)/Al(crit) ratio for Sitka spruce, compared to other conifers, and the influence of the large deposition of sea salt base cations. The low value of the (Ca + Mg + K)/Al(crit) ratio for Sitka spruce implies that it is tolerant of very acidic soil conditions, however, the ratio is based on the results of only one solution culture study and may thus be uncertain under field conditions. Large sea salt base cation deposition directly influences SMBE critical loads because the predicted soil water base cation concentrations permit large concentrations of hydrogen ions and aluminium (low ANC values) before the critical chemical limit is transgressed. Where weathering rates are low, critical ANC leaching (ANC(lecrit)) becomes the dominant term in the SMBE, with the counter intuitive result that the critical load becomes a linear function of sea salt base cation deposition. Thus the current formulation of the SMBE may not be appropriate for low weathering rate areas receiving large amounts of sea salt base cation deposition. PMID- 10843341 TI - Effects of aging on neurogenic vasodilator responses evoked by transcutaneous electrical nerve stimulation: relevance to wound healing. AB - We have previously shown an age-related decline in the modulation of skin vascular reactivity by sensory nerves that correlates with a decline in wound repair efficacy. This study was designed to examine the possibility that improving the functional ability of aged sensory nerves using noninvasive transcutaneous electrical nerve stimulation (TENS) could also accelerate tissue repair. TENS of the sciatic nerve, combined with measuring blood flow responses in the rat hind-footpad using laser Doppler flowmetry, was used to establish the vascular effects. Following TENS (using parameters 20V, 5 Hz for 1 min), similar increases in vascular responses were obtained in both young (13.2+/-0.9 cm2) and old rats (11.6+/-2.3 cm2). In contrast, capsaicin-pretreated rats showed markedly diminished responses. Sympathetic fibers did not appear to modulate these sensory nerve responses. In the second part, a thermal wound was induced (using a CO2 laser) in the interscapular region of old rats (under anesthesia). In the active treatment group, TENS was applied twice daily for the initial 5 days, and the sham group received inactive TENS. Using the healing endpoint as the time when full wound contraction occurred, the active group required 14.7+/-0.2 days for complete healing, a significant improvement over the sham group (21.8+/-0.3 days). We contend that low-frequency TENS can improve the vascular response of old rats. In addition, wound healing in aged rats can be accelerated by peripheral activation of sensory nerves at low-frequency electrical stimulation parameters. PMID- 10843342 TI - Relationships among age-associated strength changes and physical activity level, limb dominance, and muscle group in women. AB - This study investigated the magnitude and rate of age-associated strength reductions in Australian independent urban-dwelling women and the relationship to muscle groups, limb dominance, and physical activity level. Independent urban dwelling women aged 20 to 89 years (N = 217) performed maximal voluntary contractions with the dominant and nondominant knee extensors, plantar flexors, and handgrip. Anthropometric measurements were made and questionnaire responses used to obtain current physical activity levels. Trend analysis within analysis of variance and regression analysis on strength was performed. Limb muscle strength was found to be associated with increased age, muscle group, limb dominance, and activity. Self-reported physical activity levels declined with age but women who were more physically active for their age group were stronger in all muscle groups and had more lean body mass and lean thigh and leg cross sectional area than relatively inactive women. Slopes of the linear reductions of maximal voluntary strength of the knee extensors, plantar flexors, and handgrip with age were significantly different (p < .05) at 9.3%, 7.4%, and 6.2% per decade, respectively. The limb muscle strength of healthy Australian independent and urban-dwelling women aged 20 to 89 years was found to be associated with age and three aspects of disuse: muscle group, relative levels of physical activity, and limb dominance. PMID- 10843343 TI - Effects of aging and dietary restriction on mRNA levels of receptors for growth hormone-releasing hormone and somatostatin in the rat pituitary. AB - Aging impairs and dietary restriction may modulate pituitary response to growth hormone (GH)-releasing hormone (GHRH) and somatostatin (SRIH) for GH secretion. Using the semiquantitative reverse-transcription polymerase chain reaction method, we analyzed the mRNA levels of the GHRH receptor (grfr) and SRIH receptor subtype 2 (sstr2) and subtype 5 (sstr5) in anterior pituitaries of male rats fed ad libitum or 30% dietary restricted. Aging reduced the mRNA levels of these receptors in a slightly different manner. The levels of grfr progressively decreased between 6 and 24 months, whereas those of sstr2 and sstr5 declined after 16 months. Dietary restriction did not diminish the aging-dependent changes, although it slightly augmented the levels of grfr, but not sstr2 and sstr5. The present results suggest that the aging-dependent impairment in pituitary response for GH secretion could result mostly from a decline in grfr rather than relative increase of sstrs. Although DR could slightly enhance the pituitary sensitivity to GHRH, the antiaging action may be minor at the level of gene expression. PMID- 10843344 TI - Effects of tocotrienols on life span and protein carbonylation in Caenorhabditis elegans. AB - To assess the efficiency of tocotrienols against oxidative damage, we have demonstrated in a model-system nematode, Caenorhabditis elegans, that tocotrienol administration reduced the accumulation of protein carbonyl (a good indicator of oxidative damage during aging) and consequently extended the mean life span (LS), but not the maximum LS. Conversely, alpha-tocopherol acetate did not affect these parameters. As a way to evaluate the protective ability of tocotrienols against oxidative stress, the life spans of animals administrated tocotrienols before or after exposure to ultraviolet B-induced oxidative stress were measured. Ultraviolet B irradiation shortened the mean LS of animals, whereas preadministration of tocotrienols recovered the mean LS to that of unirradiated animals. Interestingly, postadministration also extended the mean LS more than that of unirradiated animals, and administration through the LS conferred greater protection. Thus, the administration of tocotrienols to animals results in a reduction of oxidative stress risks. These data indicated that tocotrienols merit further investigation as possible agents for antiaging and oxidative stress prevention. In addition, they suggest that C. elegans will continue to provide provocative clues into the mechanisms of aging. PMID- 10843345 TI - Low fatty acid unsaturation: a mechanism for lowered lipoperoxidative modification of tissue proteins in mammalian species with long life spans. AB - Carbonyl compounds generated by the nonenzymatic oxidation of polyunsaturated fatty acids react with nucleophilic groups in proteins, leading to their modification. It has not been tested whether fatty acid unsaturation is related to steady-state levels of lipoxidation-derived protein modification in vivo. A low fatty acid unsaturation, hence a low protein lipoxidation, in tissues of longevous animals would be consistent with the free radical theory of aging, because membrane lipids increase their sensitivity to oxidative damage as a function of their degree of unsaturation. To evaluate the relationship between fatty acid composition, protein lipoxidation, and maximum life span (MLSP), we analyzed liver fatty acids and proteins from seven mammalian species, ranging in MLSP from 3.5 to 46 years. The results show that the peroxidizability index of fatty acids and the sensitivity to in vitro lipid peroxidation are negatively correlated with the MLSP. Based on gas chromatography and mass spectroscopy analyses, liver proteins of all these species contain malondialdehyde-lysine and Nepsilon-carboxymethyllysine adducts, two biomarkers of protein lipoxidation. The steady-state levels of malondialdehyde-lysine and Nepsilon-carboxymethyl lysine are directly related to the peroxidizability index and inversely related to the MLSP. We propose that a low degree of fatty acid unsaturation may have been selected in longevous mammals to protect their tissue lipids and proteins against oxidative damage while maintaining an appropriate environment for membrane function. PMID- 10843346 TI - Extended longevity in Drosophila is consistently associated with a decrease in developmental viability. AB - It has proven relatively easy to select normal-lived strains of Drosophila for extended longevity in the laboratory. Long-lived strains have not been observed in the wild as yet. Of the various life-history traits that have been investigated for their role in modulating the evolution of extended longevity, none have yet shown a consistent or convincing relationship. Other than developmental time, the traits usually investigated in this regard are those associated with the adult phase of the life cycle. We assayed developmental timing and viability in six pairs of normal- and long-lived strains, four pairs of which are from previously described strains and two pairs of which are new strains that have been independently and recently selected. We find that the life history trait most obviously associated with all our long-lived strains is a significantly reduced developmental viability, with the long-lived strains' having as much as twice the developmental lethality as do any of the normal-lived strains. The long-lived strains also pupate closer to the food, a behavior known to decrease fitness. Thus the reduced fitness of the long-lived strains appears to be due to both physiological and behavioral factors and may well explain why long lived strains are not usually found in the wild. The extension of longevity involves costs as well as benefits that, in this case, are borne by different individuals. PMID- 10843347 TI - Determinants of peak V(O2) in peripheral arterial occlusive disease patients. AB - Peripheral arterial occlusive disease (PAOD) patients with intermittent claudication are functionally limited and deconditioned. This study examined whether peak aerobic capacity (V(O2) peak) was associated with PAOD severity, muscle mass, and comorbidities in 109 PAOD patients (93 men and 16 women) aged 48 86 years. The V(O2) peak (1.12+/-0.34 L/min), percentage body fat (30.6+/-8.3%), lean tissue mass of the total body (51.4+/-8.4 kg), lean tissue mass of the legs (16.6+/-3.0 kg), and appendicular skeletal mass (22.8+/-4.2 kg) were determined. The lean tissue mass of the total body (r = .44), lean tissue of the legs (r = .43) and resting ankle/brachial systolic pressure index (ABI; r = .41) correlated with peak V(O2) (all p < .001). None of the comorbidity variables (obesity, arthritis, coronary artery disease, hypertension, diabetes, and smoking history) were significantly associated with peak V(O2) except smoking status. The final model for the prediction of peak V(O2) included lean tissue mass of the legs, resting ABI, smoking status, and ABI x smoking status (r2 = .37,p < .001). In older patients with intermittent claudication, lean tissue mass is an important determinant of physical performance independent of PAOD severity and smoking status. Prevention of muscle atrophy may preserve ambulatory function and peak exercise capacity in older PAOD patients. PMID- 10843349 TI - Exercise strategies should be designed to increase muscle power. PMID- 10843348 TI - Age effects on the adaptive response of the female rat heart following aortic constriction. AB - Aging is associated with adaptations in the hearts of mammals that diminish the reserve capacity to meet hemodynamic loading challenges. To evaluate potential mechanisms of this phenomenon, the following hypotheses were tested: compared with hearts of adult rats, hearts of aged rats undergoing aortic constriction will exhibit (a) a lower concentration of myofibrillar proteins, (b) a reduced sensitivity to extracellular calcium, and (c) a reduced coronary perfusion. Female Fischer 344 rats aged 9 months (adult) and 27 months (aged) were assigned to control (C) or aortic-constriction (AC) groups and studied at 7 and 28 days post-AC, yielding six groups of rats. Analysis of variance was used to examine the effects of age and AC. The left ventricular (LV) mass/body mass ratio expressed a percentage of age-matched control value averaged AC-7adult, 111%; AC 28adult, 120%; AC-7aged, 106%; AC-28aged, 123% (AC, p < .01). As a percentage of adult rats values, the pressure-generating capacity of the LV averaged Caged, 99%; AC-7aged, 92%; AC-28aged, 92% (age, p < .05). There were no differences attributable to age or AC in either myofibrillar protein concentration or calcium sensitivity. There was, however, a significantly lower concentration of nonmyofibrillar protein (approximately 10%) in the hearts of all three groups of aged rats compared with the adult rats that was unaltered by AC. The percentages of LV myosin heavy chain in the alpha-isoform were Cadult, 77%; AC-7adult, 66%; AC-28adult, 66%; Caged, 45%; AC-7aged, 41%; AC-28aged, 32% (age, p < .01; AC,p < .01). Coronary flow per gram of tissue averaged 9% lower in all three of the aged groups compared with the adult rats and was not significantly affected by AC (age, p < .05). The data suggest that a reduction in nonmyofibrillar protein and a reduced coronary flow, rather than changes in calcium sensitivity or myofibrillar protein, are associated with an impairment in the adaptive response of the aged heart. PMID- 10843350 TI - Maximal power across the lifespan. AB - BACKGROUND: Previous investigators have reported that maximal power increases during growth and decreases with aging. These age-related differences have been reported to persist even when power is scaled to body mass or muscle size. We hypothesized that age-related differences in maximal power were primarily related to differences in muscle size and fiber-type distribution rather than to age per se. METHODS: Maximum cycling power (Pmax) and optimal pedaling rate (Vopt, a surrogate measure for muscle fiber type) were determined for 195 boys and men, 8 70 years of age, by using inertial load cycle ergometry. Anthropometric dimensions were used to estimate lean thigh volume (LTVest) of all subjects, and magnetic resonance imagery was used to determine thigh and hip muscle volume (MRIvol) for 24 subjects. RESULTS: Pmax was highly related to the product of LTVest and Vopt (LTVest X Vopt; r2 = .83). Multiple regression revealed that Pmax was significantly related to both LTVest x Vopt and age (R2 = .84). Power scaled by LTVest X Vopt was stable during growth and exhibited a small but significant decrease with aging. MRIvol was highly correlated with LTVest, and the ratio of LTVest to MRIvol was independent of age. CONCLUSIONS: These results suggest that muscle volume and optimal pedaling rate are the main determinants of maximal power across the lifespan and that the contractile properties of muscle are developed early in childhood and remain nearly intact late into the lifespan. PMID- 10843351 TI - Effects of a group exercise program on strength, mobility, and falls among fall prone elderly men. AB - OBJECTIVES: This randomized controlled trial studied the effects of a low- to moderate-intensity group exercise program on strength, endurance, mobility, and fall rates in fall-prone elderly men with chronic impairments. METHODS: Fifty nine community-living men (mean age = 74 years) with specific fall risk factors (i.e., leg weakness, impaired gait or balance, previous falls) were randomly assigned to a control group (n = 28) or to a 12-week group exercise program (n = 31). Exercise sessions (90 minutes, three times per week) focused on increasing strength and endurance and improving mobility and balance. Outcome measures included isokinetic strength and endurance, five physical performance measures, and self-reported physical functioning, health perception, activity level, and falls. RESULTS: Exercisers showed significant improvement in measures of endurance and gait. Isokinetic endurance increased 21% for right knee flexion and 26% for extension. Exercisers had a 10% increase (p < .05) in distance walked in six minutes, and improved (p < .05) scores on an observational gait scale. Isokinetic strength improved only for right knee flexion. Exercise achieved no significant effect on hip or ankle strength, balance, self-reported physical functioning, or number of falls. Activity level increased within the exercise group. When fall rates were adjusted for activity level, the exercisers had a lower 3-month fall rate than controls (6 falls/1000 hours of activity vs 16.2 falls/1000 hours, p < .05). DISCUSSION: These findings suggest that exercise can improve endurance, strength, gait, and function in chronically impaired, fall prone elderly persons. In addition, increased physical activity was associated with reduced fall rates when adjusted for level of activity. PMID- 10843352 TI - Mild cognitive impairment in the population and physical health: data on 1,435 individuals aged 75 to 95. AB - BACKGROUND: The physical health correlates of mild cognitive impairment (MCI) in the older individual are poorly known. The aim of this study was to investigate the relationship between physical health and MCI with population data. METHODS: Subjects were 1,435 nondemented 75- to 95-year-old subjects. MCI was defined as scoring one standard deviation below age- and education-specific means on the Mini-Mental State Examination. MCI was consistently associated with indicators of poorer health in logistic regression models with adjustment for potential confounders. RESULTS: The adjusted odds ratios for those with two, three, four, or more somatic symptoms compared with those with one or no symptoms were 1.3 (95% confidence intervals 1.0 to 1.9) and 2.1 (1.2 to 4.5; p for trend =.004); for those with poor self-rated health the odds ratio was 1.9 (1.4 to 2.6); for those with one, two, or more chronic diseases compared with those with no chronic diseases, the odds ratios were 1.3 (0.9 to 1.9) and 3.0 (1.2 to 7.6; p for trend =.02); and for those dying during the 3-year follow-up period the odds ratio was 1.5 (1.1 to 2.2). CONCLUSIONS: MCI is associated with poor physical health, leading to the hypothesis of a causal relationship between physical diseases and MCI in older populations. PMID- 10843353 TI - Effects of aging on cardiovascular responses to gravity-related fluid shift in humans. AB - BACKGROUND: Fluid shift induced by postural change causes autonomic neural responses of the cardiovascular system that buffer blood pressure fluctuation. The aim of the study was to clarify the effects of aging on cardiovascular autonomic functions in response to gravity-related fluid shift that unloads or loads the baroreceptors in human subjects. METHODS: A chest electrocardiogram, blood pressure by Finapres, and stroke volume by impedance method were measured in healthy young men (23-31 years old) and healthy elderly men (74-80 years old) during supine rest, at 90 degrees head-up tilt and thermoneutral head-out water immersion. Spectral analysis was applied to the time series data of the R-R intervals (heart rate variability [HRV]) and systolic blood pressure (blood pressure variability [BPV]). The arterial baroreflex gain for heart rate was estimated using frequency transfer function analysis. RESULTS: The young subjects had stable blood pressure, despite the larger amount of fluid shift induced by both tilt and immersion, and had marked changes in HRV and BPV. The elderly subjects failed to maintain stable blood pressure during these perturbations, despite less fluid shift and no significant changes in HRV and BPV. The arterial baroreflex gain for heart rate was not changed in the elderly subjects, whereas the gain decreased with upright in the young subjects and showed an increasing tendency during immersion compared with upright posture. CONCLUSIONS: These findings suggest that the adaptivity of the autonomic nervous system to gravity related fluid shift is reduced in elderly people, and this may cause blood pressure instability. PMID- 10843354 TI - Accelerated dysfunction among the very oldest-old in nursing homes. AB - BACKGROUND: The population aged 65 and older is often analyzed in three categories: young-old (65-74), middle-old (75-84), and oldest-old (> or = 85). This may blind heterogeneity within the oldest category. New, large data sets allow examination of the very oldest-old (e.g., aged > or = 95) and contrasts with those who are younger. METHODS: We determined the annual change of prevalence of physical and cognitive function, and of disease problems in the old to very oldest-old, using data from existing Resident Assessment Instrument records from nursing homes in seven states during 1992-1994. We used data from 193,467 unique residents aged 80 or older, including 6,556 residents aged 100 or older. We computed the prevalence, by age, of selected conditions: physical and cognitive function, diseases, problem behavior, mood disturbance, restraint use, falls, weight loss, eating less, body mass index, chewing and swallowing problems, incontinence (bowel and bladder), catheter use, and selected diagnoses. RESULTS: Prevalence of all measures of physical and cognitive dysfunction increased most rapidly with each year of age among the very oldest-old. Most of the slope changes occurred from 95 to 100 years of age. Such changes are less pronounced or not seen in measures of disease prevalence. CONCLUSIONS: Accelerated change in prevalence of dysfunction seen in the nursing home population may suggest a change in the mechanisms of aging that occur after the mid-nineties. Examination of the very oldest-old may provide new insight into the nature of the aging process. PMID- 10843355 TI - Cardiovascular fitness, body composition, and lipoprotein lipid metabolism in older men. AB - BACKGROUND: Lipoprotein lipids in older individuals are affected by family history of coronary artery disease (CAD), obesity, diet, and physical activity habits. METHODS: The relationship of obesity and physical fitness (VO2max) to lipoprotein lipids and postheparin lipases was examined in a cross-sectional study of 12 lean (LS) and 26 obese (OS) sedentary men and 18 master athletes (MAs) aged 65+/-1 years (mean +/- SE). The men were healthy, had no family history of CAD, and were weight stable on AHA diets at the time of study. RESULTS: VO2max was similar in LS and OS men but higher in the MAs. The OS men had a higher percentage of body fat (%BF), waist circumference, and waist:hip ratio (WHR) than the MA and LS men, but MA and LS men differed only in waist circumference. Total and LDL-C levels were comparable, but HDL-C, HDL2-C, and %HDL2b subspecies were higher in MAs than in OS and LS men, and in LS than in OS men. Triglyceride (TG) was similar in MAs and LS men but higher in OS men. Across groups, two multiple regression analyses models (VO2max, %BF, and WHR or waist circumference) showed that %BF and VO2max independently predicted HDL-C and HDL2, whereas WHR predicted TG (r2 = .45) more strongly than waist circumference (r2 = .39). Postheparin lipoprotein lipase activity (LPL) was comparable among groups and correlated independently with VO2max. Total postheparin lipolytic activity (PHLA), hepatic lipase activity (HL), and HL:PHLA ratio were similar in MAs and LS men but higher in OS men. In both multiple regression analysis models, only %BF predicted HL activity and the HL:PHLA ratio. The HL:PHLA ratio independently predicted HDL-C, HDL2-C, %HDL2b, %HDL3 subspecies, and the cholesterol:HDL-C ratio, whereas LPL activity predicted TG. CONCLUSIONS: Increased fitness and reduced total and abdominal fatness in MAs are associated with lower HL and higher LPL activities, which may mediate their higher HDL-C and lower TG levels relative to their sedentary peers. PMID- 10843356 TI - Physical and performance measures for the identification of mild to moderate frailty. AB - BACKGROUND: The relative importance and association of factors contributing to physical frailty in elderly persons are unclear. METHODS: Physical measures of upper and lower extremity strength, range of motion, balance, coordination, sensation, and gait were evaluated in relation to scores obtained on a 36-point physical performance test (PPT) in 107 elderly subjects. RESULTS: Scores on the PPT were significantly associated with the measures of strength and balance, gait, several range of motion values, and sensation. Subjects were also grouped according to score on the PPT as not frail (32-36 points), mildly frail (25-31 points), or moderately frail (17-24 points). ANOVA followed by Bonferroni post hoc analyses were used to examine the relationships of physical measures to this index of frailty. Balance measures, an obstacle course, the Berg scale, the full tandem portion of the Romberg test, and fast gait speed were significantly different among the three groups. Multiple stepwise regression analyses indicated that the strongest combination of variables, explaining 73% of all the variance in the PPT, included obstacle course performance, hip abduction strength, the semitandem portion of the Romberg test, and coordination (pegboard). CONCLUSIONS: Results provide further insight into the relative importance of factors that contribute to frailty and factors that should be considered in treatment planning for the remediation of physical frailty in old adults. PMID- 10843357 TI - Healing of diabetic foot ulcers and pressure ulcers with human skin equivalent: a new paradigm in wound healing. AB - HYPOTHESIS: In patients with diabetic foot and pressure ulcers, early intervention with biological therapy will either halt progression or result in rapid healing of these chronic wounds. DESIGN: In a prospective nonrandomized case series, 23 consecutive patients were treated with human skin equivalent (HSE) after excisional debridement of their wounds. SETTING: A single university teaching hospital and tertiary care center. PATIENTS AND METHODS: Twenty-three consecutive patients with a total of 41 wounds (1.0-7.5 cm in diameter) were treated with placement of HSE after sharp excisional debridement. All patients with pressure ulcers received alternating air therapy with zero-pressure alternating air mattresses. MAIN OUTCOME MEASURE: Time to 100% healing, as defined by full epithelialization of the wound and by no drainage from the site. RESULTS: Seven of 10 patients with diabetic foot ulcers had complete healing of all wounds. In these patients 17 of 20 wounds healed in an average of 42 days. Seven of 13 patients with pressure ulcers had complete healing of all wounds. In patients with pressure ulcers, 13 of 21 wounds healed in an average of 29 days. All wounds that did not heal in this series occurred in patients who had an additional stage IV ulcer or a wound with exposed bone. Twenty-nine of 30 wounds that healed did so after a single application of the HSE. CONCLUSIONS: In diabetic ulcers and pressure ulcers of various durations, the application of HSE with the surgical principles used in a traditional skin graft is successful in producing healing. The high success rate with complete closure in these various types of wounds suggests that HSE may function as a reservoir of growth factors that also stimulate wound contraction and epithelialization. If a wound has not fully healed after 6 weeks, a second application of HSE should be used. If the wound is not healing, an occult infection is the likely cause. All nonischemic diabetic foot and pressure ulcers that are identified and treated early with aggressive therapy (including antibiotics, off-loading of pressure, and biological therapy) will not progress. PMID- 10843358 TI - What prognostic factors are important in duodenal adenocarcinoma? AB - HYPOTHESIS: Survival of patients with adenocarcinoma of the duodenum depends on the ability to perform a complete resection and the tumor stage DESIGN: Retrospective case series. SETTING: Tertiary care referral center. PATIENTS: A cohort of 101 consecutive patients (mean age, 62 years), undergoing surgery for duodenal adenocarcinoma from January 1, 1976, through December 31, 1996. Patients with ampullary carcinoma were specifically excluded. Mean duration of follow-up was 4 years. INTERVENTIONS: Surgery was curative in 68 patients (67%) and palliative in 33 patients (33%). Of the curative group, 50 patients (74%) underwent radical surgery, ie, 30 (60%), pancreaticoduodenectomy; 15 (30%), pylorus-preserving pancreaticoduodenectomy; and 5 (10%), total pancreatectomy. A more limited resection procedure was used in 18 patients (26%) involving a segmental duodenal resection in 15 (83%) and a transduodenal excision in 3 (17%). patient survival, and correlation with patient and tumor variables using univariate and multivariate analysis. RESULTS: Actuarial 5-year survival for the curative group was 54%. Only 1 patient in the unresected group survived beyond 3 years. Nodal metastasis (P = .002), advanced tumor stage (P<.001), positive resection margin (P = .02), and weight loss (P<.001) had a significant negative impact on survival in multivariate analysis. Tumor grade, size, and location within the duodenum had no impact on survival. Patient age and tumor depth of invasion influenced survival in univariate analysis, but lost their prognostic significance in multivariate analysis. CONCLUSIONS: Metastasis to lymph nodes, advanced tumor stage, and positive resection margins are associated with decreased survival in patients with duodenal adenocarcinoma. An aggressive surgical approach that achieves complete tumor resection with negative margins should be pursued. Pancreaticoduodenectomy is usually required for cancers of the first and second portion of the duodenum. Segmental resection may be appropriate for selected patients, especially for tumors of the distal duodenum. PMID- 10843359 TI - The increasing problem of unusual pancreatic tumors. AB - HYPOTHESIS: Patients presenting with a pancreatic mass often have a curable lesion rather than the more common adenocarcinoma. Greater awareness of this among nonsurgeons is necessary. DESIGN: Retrospective case series. SETTING: Tertiary care referral hospital. PATIENTS: All patients who presented with a pancreatic mass during the 8 years from 1990 to 1998 were studied. Patients with a history of chronic pancreatitis, a functioning pancreatic neuroendocrine tumor, or pancreatic adenocarcinoma were excluded. Forty patients were identified, demographic and clinical characteristics recorded, and long-term follow-up obtained. INTERVENTIONS: Therapy included either a Whipple procedure or distal pancreatectomy. Two patients underwent a biliary bypass. MAIN OUTCOME MEASURES: Tumor histology, morbidity, and survival. RESULTS: Three hundred thirty-six patients with a pancreatic mass were treated during this 8-year period. Two hundred ninety-six of these had pancreatic adenocarcinoma. Forty (11.9%) of the 336 patients had other types of pancreatic tumors. Two thirds of these patients were female, with an average age of 57 years. Seventy-five percent of these tumors were either malignant or potentially malignant. In several instances, cystic tumors were diagnosed as inflammatory pseudocysts and managed accordingly. Fourteen (35%) of 40 patients had no symptoms and their tumor was found on a computed tomographic scan performed for another indication. Percutaneous biopsy was performed in 9 patients, of whom 5 were assigned an incorrect diagnosis. There were no operative deaths, although the postoperative complication rate was 23%. CONCLUSIONS: In this series, nearly 12% of patients presenting with a pancreatic mass did not have pancreatic adenocarcinoma, but rather more favorable lesions amenable to operation. Preoperative biopsy should not be carried out. Curative procedures can be safely performed in centers seeing a large number of patients with pancreatic tumors, and the long-term results of extirpation are excellent. PMID- 10843360 TI - Determinants of intestinal metaplasia within the columnar-lined esophagus. AB - HYPOTHESIS: The clinical and physiological features of patients with short segments of columnar-lined esophagus (CLE) with and without intestinal metaplasia (IM) are distinct. DESIGN: Retrospective case series. SETTING: University tertiary referral center. PATIENTS: Sixty-five consecutive patients with a 2-cm or shorter length of endoscopically visible CLE. INTERVENTIONS: The type of CLE and the presence of Helicobacter pylori were determined by histopathologic examination of esophageal and gastric antrum biopsy specimens. All patients underwent esophageal manometry and simultaneous 24-hour pH and bilirubin monitoring. MAIN OUTCOME MEASURES: Clinical and physiological data were compared in patients with and without IM. RESULTS: Thirty-six patients had IM and 29 had cardiac-type mucosa without IM in biopsy specimens from the CLE. There was no significant difference in age or sex distribution, but the duration of symptoms was significantly longer in patients with IM (10 vs 5 years; P = .03). Abnormal esophageal acid exposure was found in 30 (83%) of 36 patients with IM and 23 (79%) of 29 patients without IM. The prevalence of abnormal bilirubin exposure was significantly higher in patients with IM (75% [27/36]) than in those without IM (41% [12/29]; P = .01). There was no significant difference in the prevalence of H pylori infection between the 2 groups (8% vs 10%; P >.99). CONCLUSIONS: Patients with short segments of CLE and IM have similar esophageal acid exposure but significantly higher frequency of abnormal bilirubin exposure and longer median duration of reflux symptoms than patients without IM. Therefore, CLE, regardless of histological type, is a manifestation of gastroesophageal reflux disease. The presence of duodenoesophageal reflux and the duration of reflux seem to be important in the pathogenesis of IM. PMID- 10843361 TI - Cryosurgical ablation and radiofrequency ablation for unresectable hepatic malignant neoplasms: a proposed algorithm. AB - BACKGROUND: Thermal ablation of unresectable hepatic tumors can be achieved by cryosurgical ablation (CSA) or radiofrequency ablation (RFA). The relative advantages and disadvantages of each technique have not yet been determined. HYPOTHESIS: Radiofrequency ablation of malignant hepatic neoplasms can be performed safely, but is currently limited by size. Cryosurgical ablation, while associated with higher morbidity, is more effective for larger unresectable hepatic malignant neoplasms. DESIGN: Retrospective analysis of prospective patient database. PATIENTS AND METHODS: Between July 1992 and September 1999, 308 patients with liver tumors not amenable to curative surgical resection were treated with CSA and/or RFA (percutaneous, laparoscopic, celiotomy). No patient had preoperative evidence of extrahepatic disease. All patients underwent laparoscopy with intraoperative ultrasound if technically possible. Both RFA and CSA were performed under ultrasound guidance. Resection, as an adjunctive procedure, was combined with ablation in certain patients. RESULTS: Laparoscopy identified extrahepatic disease in 12% of patients, and intraoperative hepatic ultrasound identified additional lesions in 33% of patients, despite extensive preoperative imaging. Radiofrequency ablation alone or combined with resection or CSA resulted in reduced blood loss (P<.05), thrombocytopenia (P<.05), and shorter hospital stay compared with CSA alone (P<.05). Median ablation times for lesions greater than 3 cm were 60 minutes with RFA and 15 minutes with CSA (P<.001). Local recurrence rates for lesions greater than 3 cm were also greater with RFA (38% vs 17%). CONCLUSIONS: Laparoscopy and intraoperative ultrasound are essential in staging patients with hepatic malignant neoplasms. Radiofrequency ablation when combined with CSA reduces the morbidity of multiple freezes. Although RFA is safer than CSA and can be performed via different approaches (percutaneously, laparoscopically, or at celiotomy), it is limited by tumor size (<3 cm). Percutaneous RFA should be considered in high-risk patients or those with small local recurrences. PMID- 10843362 TI - Hepatic cryoablation-induced acute lung injury: pulmonary hemodynamic and permeability effects in a sheep model. AB - HYPOTHESIS: Hepatic cryoablation of 30% to 35% or more of liver parenchyma in a sheep model results in eicosanoid and nuclear factor-kappaB (NF-kappaB)-mediated changes in pulmonary hemodynamics and lung permeability. SETTING: Laboratory. INTERVENTIONS: At initial thoracotomy, catheters were placed in the main pulmonary artery, left atrium, right carotid artery, and efferent duct of the caudal mediastinal lymph node for subsequent monitoring in adult sheep. After a 1 to 2-week period of recovery, animals underwent laparotomy and left-lobe cryoablation (approximately 35% by volume) with subsequent awake monitoring and on postoperative days 1 to 3. MAIN OUTCOME MEASURES: Cryoablation-induced lung permeability and hemodynamic changes were compared with baseline values in sheep that underwent instrumentation. Similarly handled sheep underwent resection of a similar volume of hepatic parenchyma or had pulmonary artery pressure increases induced by mechanical left atrial obstruction. Activation of NF-kappaB was assessed with electrophoretic mobility shift assay, and serum thromboxane levels were measured with mass spectroscopy. RESULTS: Cryoablation resulted in acutely increased mean pulmonary (20 to 35 cm water) and systemic pressures, which returned to baseline at 24 hours with no change in cardiac output. Serum thromboxane levels increased 30 minutes after cryoablation (9-fold) and returned to baseline at 24 hours. Activation of NF-kappaB was present in liver and lung tissue by 30 minutes after cryoablation. Lung lymph-plasma protein clearance markedly exceeded the expected increase from pulmonary pressures alone, and increased lymph-plasma protein ratio persisted after pulmonary artery pressures normalized. Similar changes were not associated with 35% hepatic resection. CONCLUSIONS: This study demonstrates that 35% hepatic cryoablation results in an acute but transient increase in pulmonary artery pressure that may be mediated by increased thromboxane levels. Increases in pulmonary capillary permeability are not accounted for by pressure changes alone, and may be a result of NF-kappaB mediated inflammatory mechanisms. These data show that cryosurgery causes pathophysiological changes similar to those observed with endotoxin and other systemic inflammatory stimuli. PMID- 10843363 TI - Nonoperative management of splenic injuries: have we gone too far? AB - HYPOTHESIS: Patients with severe blunt injuries to the spleen have a high likelihood of failing nonoperative management of splenic injuries (NOMSI). DESIGN: Review of medical records, helical computed tomographic imaging data, and trauma registry data. SETTING: Academic level I trauma center at a large county hospital. PATIENTS: A total of 105 patients with blunt trauma to the spleen, admitted between January 1995 and December 1998, who survived more than 48 hours and had complete records. Of these patients, 53 (56%) were selected for NOMSI. The splenic injury was graded by the Organ Injury Scale of the American Association for the Surgery of Trauma (grades I to V, with grade V being the worst possible injury). MAIN OUTCOME MEASURES: Failure of NOMSI, defined as the need for operation to the spleen after a period of nonoperative management. RESULTS: Compared with patients who had successful NOMSI, the 29 patients (52%) in whom NOMSI failed were older and more severely injured. They also required extra-abdominal operations more frequently, underwent transfusion with more units of blood while being managed nonoperatively, and had higher grades of splenic injury. Splenic injury grade III or higher and transfusion of more than 1 U of blood were identified as independent risk factors for failure of NOMSI. The existence of both risk factors predicted failure in 97% of cases. The grading by computed tomography correlated well with the actual injury to the spleen as seen at operation. CONCLUSIONS: In patients with high-grade splenic injuries who require a transfusion of more than 1 U of blood, NOMSI is very likely to fail. Decreasing the threshold for operation or intensifying the monitoring is highly recommended for such patients. PMID- 10843364 TI - Ogilvie syndrome as a postoperative complication. AB - HYPOTHESIS: Ogilvie syndrome is a postoperative complication. DESIGN: Case series. SETTING: University-affiliated tertiary-care hospital. PATIENTS AND METHODS: The medical records of patients diagnosed as having Ogilvie syndrome after trauma or operation between 1989 and 1998 were reviewed. Medical charts were examined for history, treatment, cecal diameter, and outcome. MAIN OUTCOME MEASURES: Data were summarized in an attempt to identify patient populations at risk for Ogilvie syndrome. RESULTS: Ogilvie syndrome was diagnosed in 36 patients, 24 of whom were men. Average age at diagnosis was 68.9 years. Abdominal radiographs were obtained at time of diagnosis (mean cecal diameter, 13.4 cm; range, 8-20 cm). Operations preceding Ogilvie syndrome were orthopedic or spinal (n= 14), cardiothoracic (n= 12), abdominal (n= 5), and vascular (n= 2). Nonoperative trauma accounted for 3 cases. Coronary artery bypass grafting was the single most frequent procedure leading to Ogilvie syndrome (n=9 [25%]). Conservative treatment was successful in 52.8% of cases (n = 19). Twenty colonoscopic decompressions were performed on 13 patients, with an overall success rate of 77% (n= 10). Of the 3 patients in whom colonoscopic decompression failed, 2 died and 1 required operation. Five of the 36 patients required surgical intervention, with a mortality rate of 60% (n= 3). CONCLUSIONS: Previous studies have shown Ogilvie syndrome to occur most commonly after obstetrical/ gynecologic, abdominal/pelvic, and orthopedic procedures. Our data confirm that patients undergoing orthopedic and spinal procedures are at higher risk, but that the surgical procedure most commonly leading to Ogilvie syndrome was coronary artery bypass grafting. Cardiothoracic surgeons, orthopedic surgeons, and neurosurgeons should be cognizant of this complication in the patient whose abdomen becomes distended postoperatively. If recognized early and treated appropriately, pseudo-obstruction will resolve in most patients. If surgical intervention is required, the subsequent mortality rate is high. PMID- 10843365 TI - Blunt trauma resuscitation: the old can respond. AB - HYPOTHESIS: Old and young trauma patients are capable of hyperdynamic response during standardized shock resuscitation. DESIGN: The responses of old and young trauma patients resuscitated using a standardized protocol are compared in an inception cohort study. A standardized resuscitation protocol was used to attain and maintain an oxygen delivery index of 600 mL/min x m2 or greater (DO2I > or = 600) for the first 24 hours in the intensive care unit. Interventions, responses, and outcomes for old (> or = 65 years) and young (<65 years) patients are described. Data were analyzed using analysis of variance, the chi2 test, and the t test; P<.05 was considered significant. SETTING: A 20-bed shock trauma intensive care unit in a regional level I trauma center. PATIENTS: Patients at high risk of postinjury multiple organ failure, ie, major organ or vascular injury and/or skeletal fractures, initial base deficit of 6 mEq/L or greater, need for 6 units or more of packed red blood cells in the first 12 hours, or age of 65 years or older with any 2 previous criteria. INTERVENTIONS: Pulmonary artery catheter, crystalloid fluid infusion, packed red blood cell transfusion, and moderate inotrope support, as needed in that sequence, to attain DO2I > or = 600. MAIN OUTCOME MEASURES: Intensive care unit length of stay and survival. RESULTS: During 19 months ending June 1999, 12 old patients (58% male; age, 76 +/ 2 years [mean +/- SEM] [P<.0011; Injury Severity Score, 20 +/- 2 [P=.02]) and 54 young patients (61% male; age, 37 +/- 2 years; Injury Severity Score, 32 +/- 2) were resuscitated. Initially, for old patients (cardiac index, 2.0 +/- 0.2 L/min x m2) and for young patients (cardiac index, 3.0 +/- 0.2 L/min x m2; P=.01), 24 hour volumes were as follows: 16 +/- 3 L of crystalloid and 12 +/- 3 units of packed red blood cells for the old patients and 21 +/- 2 L of crystalloid and 19 +/- 2 units of packed red blood cells for the young patients. For old patients, 9 (75%) attained DO2I > or = 600, and 11 (92%) survived 7 or more days and 5 (42%) 30 or more days. For young patients, 45 (83%) attained the DO2I goal, and 48 (89%) survived 30 or more days. Intensive care unit length of stay was 25 +/- 9 days for the old patients and 23 +/- 2 days for the young patients. CONCLUSIONS: Elderly patients have initially depressed cardiac index but generate hyperdynamic response. Although ultimate outcome is poorer than in the younger cohort, resuscitation is not futile. PMID- 10843366 TI - Computer-aided diagnosis for surgical office-based breast ultrasound. AB - HYPOTHESIS: The computer-aided diagnostic system is an intelligent system with great potential for categorizing solid breast nodules. It can be used conveniently for surgical office-based digital ultrasonography (US) of the breast. DESIGN: Retrospective, nonrandomized study. SETTING: University teaching hospital. PATIENTS: We retrospectively reviewed 243 medical records of digital US images of the breast of pathologically proved, benign breast tumors from 161 patients (ie, 136 fibroadenomas and 25 fibrocystic nodules), and carcinomas from 82 patients (ie, 73 invasive duct carcinomas, 5 invasive lobular carcinomas, and 4 intraductal carcinomas). The digital US images were consecutively recorded from January 1, 1997, to December 31, 1998. INTERVENTION: The physician selected the region of interest on the digital US image. Then a learning vector quantization model with 24 autocorrelation texture features is used to classify the tumor as benign or malignant. In the experiment, 153 cases were arbitrarily selected to be the training set of the learning vector quantization model and 90 cases were selected to evaluate the performance. One experienced radiologist who was completely blind to these cases was asked to classify these tumors in the test set. MAIN OUTCOME MEASURE: Contribution of breast US to diagnosis. RESULTS: The performance comparison results illustrated the following: accuracy, 90%; sensitivity, 96.67%; specificity, 86.67%; positive predictive value, 78.38%; and negative predictive value, 98.11% for the computer-aided diagnostic (CAD) system and accuracy, 86.67%; sensitivity, 86.67%; specificity, 86.67%; positive predictive value, 76.47%; and negative predictive value, 92.86% for the radiologist. CONCLUSION: The proposed CAD system provides an immediate second opinion. An accurate preoperative diagnosis can be routinely established for surgical office-based digital US of the breast. The diagnostic rate was even better than the results of an experienced radiologist. The high negative predictive rate by the CAD system can avert benign biopsies. It can be easily implemented on existing commercial diagnostic digital US machines. For most available diagnostic digital US machines, all that would be required for the CAD system is only a personal computer loaded with CAD software. PMID- 10843367 TI - Vacuum-assisted stereotactic breast biopsy: histologic underestimation of malignant lesions. AB - HYPOTHESIS: The histopathologic correlation between stereotactic core needle biopsy and subsequent surgical excision of mammographically detected nonpalpable breast abnormalities is improved with a larger-core (11-gauge) device. DESIGN: Retrospective medical record and histopathologic review. SETTING: University based academic practice setting. PATIENTS: Two hundred one patients who underwent surgical excision of mammographic abnormalities that had undergone biopsy with an 11-gauge vacuum-assisted stereotactic core biopsy device. MAIN OUTCOME MEASURE: Correlation between stereotactic biopsy histologic results and the histologic results of subsequent surgical specimens. RESULTS: Results of stereotactic biopsy performed on 851 patients revealed atypical hyperplasia in 46 lesions, ductal carcinoma in situ (DCIS) in 89 lesions, and invasive cancer in 73 mammographic abnormalities. Subsequent surgical excision of the 46 atypical lesions revealed 2 cases of DCIS (4.3%) and 4 cases of invasive carcinoma (8.7%). Lesions diagnosed as DCIS on stereotactic biopsy proved to be invasive carcinoma in 10 (11.2%) of 89 patients on subsequent excision. Stereotactic biopsy completely removed 21 (23.6%) of 89 DCIS lesions and 20 (27.4%) of 73 invasive carcinomas. CONCLUSIONS: In summary, 11-gauge vacuum-assisted core breast biopsy accurately predicts the degree of disease in the majority of malignant lesions; however, understaging still occurs in 11% to 13% of lesions showing atypical hyperplasia or DCIS. PMID- 10843368 TI - Resectional treatment for thyroid cancer with tracheal invasion: a long-term follow-up study. AB - HYPOTHESIS: Transmural invasion of the trachea by well-differentiated thyroid carcinomas is a locally advanced disease condition. It frequently causes deaths owing to airway obstruction. We hypothesized that resection of the invaded trachea followed by primary anastomosis provides the opportunity for cure. DESIGN: A retrospective review study of medical records. SETTING: The surgical department of a tertiary referral center. PATIENTS: Eight patients with well differentiated thyroid carcinomas, complicated with tracheal invasion resulting in bleeding and airway obstruction, operated on by tracheal resection and immediate anastomosis, were included. INTERVENTIONS: All patients received total thyroidectomy and neck lymph node dissection as well as segmental tracheal resection followed by primary reconstruction. Postoperative radioactive sodium iodine I 131 treatment and suppression therapy with thyroxine were applied to all of them. MAIN OUTCOME MEASURE: Postoperative follow-up of serum levels of human thyroglobulin and abnormal radioactive iodine 131I from whole body scan. RESULTS: Seven patients are alive. Of these patients, 5 had no evidence of disease: (3 had no evidence of cancer for more than 10 years' follow-up), 2 had regional lymph node metastasis, and 1 had lung metastases. The remaining patient had anastomotic site recurrence with airway obstruction and needed tracheostomy to relieve stridor. She was lost to follow-up 39 months after undergoing the initial operation. CONCLUSIONS: Patients with mucosal invasion of the trachea by well differentiated thyroid carcinomas should be treated by surgical resection followed by primary reconstruction when technically feasible. This facilitates postoperative care, and it is possible to achieve long-term survival with improvement of the quality of life and possible cures. PMID- 10843369 TI - Surgeon compensation and motivation. AB - HYPOTHESIS: Financial incentives are the only form of compensation that will motivate surgeons at an academic health sciences center to perform the tasks outlined in the hospital's mission statement. DESIGN: A questionnaire divided into 5 sections: demographics, compensation, time allocation, benefits and incentives, and motivational factors. SETTING: The Department of Surgery, The Toronto Hospital, Toronto, Ontario. PARTICIPANTS: All academic surgeons (N=64) practicing at The Toronto Hospital in July 1997. RESULTS: Of the 64 eligible mailed surveys, there were responses for 59. Of these 59 surgeons, 48 (81%) receive compensation through a fee-for-service method. However, only 32 (54%) of the surgeons prefer the fee-for-service method, while 18 (31%) prefer salary and 9 (15%) prefer an alternative system. On average, these academic surgeons spend 44% of their time teaching or performing research, for which they receive 14% of their total income. Of the motivational factors assessed, financial bonuses are a positive motivational factor for all "surgeon tasks." In addition, task-specific motivational factors were established for research, teaching, and operating, including research facilities, mentorship and prestige, and interesting case types, respectively. CONCLUSIONS: Surgeons are not appropriately renumerated for time spent on academic activities, and many would prefer an alternative form of compensation to the fee-for-service method. Knowledge that surgeons are receptive to tasks supporting the hospital's mission statement leads us to conclude that appropriate motivation can shape the activity of academic surgeons. Financial rewards ranked the highest as a motivational factor for all surgeon tasks; however, task-specific motivational factors were identified. Overall, multiple factors, specifically targeted to the individual, will serve to motivate. Thus, compensation packages based on individual preferences and personal motivational factors will be the most successful. PMID- 10843370 TI - Intussusception following the Ladd procedure. AB - HYPOTHESIS: The Ladd procedure for malrotation predisposes children to postoperative intussusception (POI). DESIGN: Retrospective case-control review. SETTING: University-affiliated tertiary care pediatric hospital. PATIENTS: Five of 159 patients undergoing the Ladd procedure between 1995 and 1998 developed POI. Predisposing factors were sought by comparison with age-matched controls who underwent the Ladd procedure during the same period. The entire Ladd group was compared with all 1717 patients undergoing any other laparotomy during the same period for incidence of POI. MAIN OUTCOME MEASURES: Differences in weight, percentile weight, age, length of nasogastric suction, time to oral intake, and length of stay between Ladd patients developing POI and age-matched controls from the Ladd group were compared using the Mann-Whitney U test. Incidence of POI after the Ladd procedure and "other laparotomy" was compared using chi2 analysis. RESULTS: In the Ladd group, there were 5 cases of POI (3.1%). There was 1 case of POI (0.05%) after all other laparotomies (P<.001). Symptoms developed at a mean +/- SD of 7.2 +/- 2.1 days. Upper gastrointestinal tract with small bowel follow through showed partial bowel obstruction in 4 cases and was normal in 1 case. Reexploration took place at a mean +/- SD of 9.2 +/- 2.8 days. Children developing POI after undergoing the Ladd procedure were less likely to be small for their age (P= .03) than age-matched controls undergoing the Ladd procedure. CONCLUSIONS: The Ladd procedure predisposes children to POI. Aggressive investigation, including reexploration, should not be delayed if a child has symptoms of prolonged ileus within 2 weeks after undergoing a Ladd procedure. PMID- 10843372 TI - Out of Africa. PMID- 10843371 TI - Trauma and trauma care systems: in the throes of an identity crisis: a plea for changing the vernacular and the mind-set. PMID- 10843373 TI - Multiple arterial thromboembolisms in a patient with the 20210 A prothrombin gene mutation. AB - The most recently characterized genetic defect contributing to venous thrombophilia is the 20210 A prothrombin gene mutation. We describe a patient with this defect who had arterial thrombosis resulting in considerable mesenteric ischemia. Several environmental factors, which might otherwise be considered of low thrombotic risk, may also have contributed to her condition. The recognition of the potential for novel presentations of hypercoagulable states may contribute to a reduction in the morbidity associated with acute mesenteric ischemia. PMID- 10843374 TI - A stepwise approach to laparoscopic Nissen fundoplication: avoiding technical pitfalls. AB - Laparoscopic Nissen fundoplication is now widely used in the surgical management of gastroesophageal reflux disease. However, it is a complex operation that requires advanced laparoscopic skills. The learning curve is steep, and complications are directly related to the surgeon's experience level. Both experimental and clinical data demonstrate a decline in complications with increasing experience. We divided this complex procedure into logical and orderly components, to facilitate the learning process. We believe that this approach will not only reduce complications by highlighting potential problems at each stage but also make it easier to teach others. PMID- 10843376 TI - We missed the cut. PMID- 10843375 TI - Surgery in Hungary. AB - After providing data on the geography and demographics of Hungary, the 1100 years of Hungarian history are briefly surveyed. The introduction, development, and present state of medical education and, in particular, surgical education, including specialty training, are discussed. Attention is devoted to the organization of the surgical community and of surgical manpower in the country. Various disease patterns and their influence on surgical practice are discussed. Attention is also devoted to health care provision and research activity. Finally, the future of surgery, especially general surgery, is outlined. PMID- 10843377 TI - William Gibson (1788-1868). PMID- 10843378 TI - Antigen-specific regulation of T cell-mediated cytokine production. PMID- 10843379 TI - The temporal importance of TNFalpha expression in the development of diabetes. AB - The inflammatory cytokine tumor necrosis factor alpha (TNFalpha) has been linked to the development of several autoimmune diseases. By adapting the tetracycline regulated gene transcription system, we generated a murine model where islet specific expression of TNFalpha could be repressed/derepressed within 48 hr following introduction/removal of tetracycline in the drinking water. Here we describe the temporal importance of TNFalpha in diabetes development in mice expressing islet-specific B7-1 and TNFalpha. We show that the duration of TNFalpha-mediated inflammation, not the putative maturity of the immune system at the time of TNFalpha expression, determines diabetes progression. Further, we have described an interval between 21 and 25 days following initiation of TNFalpha expression where the fate of islet-reactive T cells is decided. PMID- 10843380 TI - BLC expression in pancreatic islets causes B cell recruitment and lymphotoxin dependent lymphoid neogenesis. AB - CXCR5, the receptor for B lymphocyte chemoattractant (BLC), is required for normal development of Peyer's patches, inguinal lymph nodes, and splenic follicles. To test the in vivo activity of BLC in isolation of other lymphoid organizers, transgenic mice were generated expressing BLC in the pancreatic islets. In addition to attracting B cells, BLC expression led to development of lymph node-like structures that contained B and T cell zones, high endothelial venules, stromal cells, and the chemokine SLC. Development of these features was strongly dependent on B lymphocytes and on lymphotoxin alpha1beta2 and could be reversed by blocking lymphotoxin alpha1beta2. These findings establish that BLC is sufficient to activate a pathway of events leading to formation of organized lymphoid tissue. PMID- 10843381 TI - IP-10 is critical for effector T cell trafficking and host survival in Toxoplasma gondii infection. AB - The generation of an adaptive immune response against intracellular pathogens requires the recruitment of effector T cells to sites of infection. Here we show that the chemokine IP-10, a specific chemoattractant for activated T cells, controls this process in mice naturally infected with Toxoplasma gondii. Neutralization of IP-10 in infected mice inhibited the massive influx of T cells into tissues and impaired antigen-specific T cell effector functions. This resulted in >1000-fold increase in tissue parasite burden and a marked increase in mortality compared to control antibody-treated mice. These observations suggest that IP-10 may play a broader role in the localization and function of effector T cells at sites of Th1 inflammation. PMID- 10843382 TI - CCR6 mediates dendritic cell localization, lymphocyte homeostasis, and immune responses in mucosal tissue. AB - Chemokine-directed migration of leukocyte subsets may contribute to the qualitative differences between systemic and mucosal immunity. Here, we demonstrate that in mice lacking the chemokine receptor CCR6, dendritic cells expressing CD11c and CD11b are absent from the subepithelial dome of Peyer's patches. These mice also have an impaired humoral immune response to orally administered antigen and to the enteropathic virus rotavirus. In addition, CCR6( /-) mice have a 2-fold to 15-fold increase in cells of select T lymphocyte populations within the mucosa, including CD4+ and CD8+ alphabeta-TCR T cells. By contrast, systemic immune responses to subcutaneous antigens in CCR6(-/-) mice are normal. These findings demonstrate that CCR6 is a mucosa-specific regulator of humoral immunity and lymphocyte homeostasis in the intestinal mucosa. PMID- 10843383 TI - Expression of intestine-specific antigen reveals novel pathways of CD8 T cell tolerance induction. AB - Reactivity to intestinal epithelium-specific antigen was studied by transgenic expression of cytosolic ovalbumin controlled by an enterocyte-specific promoter. Transferred OVA-specific CD8 cells (OT-I) preferentially expanded in mucosal lymphoid tissues and the epithelium but failed to cause tissue damage. In contrast, concomitant VSV-ova infection induced OT-I-mediated epithelial cell destruction that correlated with antigen density. OT-I cells retained in the epithelium exhibited high levels of lytic activity but were unable to produce cytokines. The mice were systemically tolerant to OVA since endogenous CD8 cells were nonresponsive to VSV-ova infection. Thus, intestinal antigen gained access to peripheral tissues via absorption from effete epithelial cells. This system demonstrated a requirement for inflammation to drive pathogenic autoreactivity against enterocytes and identified pathways of intestine-specific immunoregulation. PMID- 10843384 TI - A second amplifier function for the allergy-associated Fc(epsilon)RI-beta subunit. AB - Genetics studies have identified the gene for the high-affinity IgE receptor (FC(epsilon)RI) beta subunit as a candidate gene for atopy. We have shown that beta is an intrinsic signaling amplifier leading to enhanced allergic responses in vivo. Here we report that beta has a second amplification function: the amplification of Fc(epsilon)RI cell surface expression. This function is due to an early association of beta with alpha, resulting in improved trafficking and maturation of alpha and receptor complexes. These data provide a possible molecular explanation for the large difference in Fc(epsilon)RI density between beta-cells such as monocytes, dendritic cells, and beta+ effector cells (mast cells, basophils). In beta+ cells, the combined signaling and expression amplification results in an estimated 12- to 30-fold amplification of downstream events. PMID- 10843385 TI - LAT is essential for Fc(epsilon)RI-mediated mast cell activation. AB - The linker molecule LAT is a substrate of the tyrosine kinases activated following TCR engagement of T cells. LAT is also expressed in platelets, NK, and mast cells. Although LAT-deficient mice contain normal numbers of mast cells, we found that LAT-deficient mice were resistant to IgE-mediated passive systemic anaphylaxis. LAT-deficient bone marrow-derived mast cells (BMMC) showed normal growth and development. Whereas tyrosine phosphorylation of Fc(epsilon)RI, Syk, and Vav was intact in LAT-deficient BMMCs following Fc(epsilon)RI engagement, tyrosine phosphorylation of SLP-76, PLC-gamma1, and PLC-gamma2 and calcium mobilization were dramatically reduced. LAT-deficient BMMCs also exhibited profound defects in activation of MAPK, degranulation, and cytokine production after Fc(epsilon)RI cross-linking. These results show that LAT plays a critical role in Fc(epsilon)RI-mediated signaling in mast cells. PMID- 10843386 TI - Inducible expression of a p56Lck transgene reveals a central role for Lck in the differentiation of CD4 SP thymocytes. AB - The T lymphocyte-specific protein tyrosine kinase p56lck (Lck) is an essential component of the TCR-mediated signal transduction complex. Lck knockout mice have reduced numbers of double-positive thymocytes and very few mature single-positive cells, particularly of the CD4 lineage. Here we demonstrate the ability of a tetracycline-based tissue-specific inducible Lck transgene to restore expansion of early thymocytes and maturation of single-positive cells in Lckneg mice upon induction with doxycycline. Restoration of Lck expression is particularly important for positive selection to the CD4+ lineage but has a lesser impact on selection to the CD8+ lineage, suggesting activation of Lck is an important component of the signals involved in lineage choice during thymic differentiation. PMID- 10843387 TI - Junctional biases in the naive TCR repertoire control the CTL response to an immunodominant determinant of HSV-1. AB - The enormous diversity of the T cell pool makes it difficult to determine whether inherent biases in the naive TCR repertoire can influence T cell responsiveness. In C57BL/6 mice the cytotoxic T lymphocyte response to an immunodominant HSV-1 determinant (gB) is characterized by a prominent bias in Vbeta element usage, associated with a conserved and preferentially D element-encoded CDR3 sequence. Comparison of naive and gB-specific T cell populations revealed a similar enrichment of germline D element-encoded CDR3 sequences in the preimmune repertoire. Strikingly, eliminating the germline coding of the gB-specific CDR3 sequence caused an almost complete loss of the dominant subset of gB-specific T cells, illustrating that CDR3 biases can significantly alter both the composition and strength of an immune response. PMID- 10843388 TI - Rac2 stimulates Akt activation affecting BAD/Bcl-XL expression while mediating survival and actin function in primary mast cells. AB - Mast cells generated from Rac2-deficient (-/-) mice demonstrated defective actin based functions, including adhesion, migration, and degranulation. Rac2(-/-) mast cells generated lower numbers and less mast cell colonies in response to growth factors and were deficient in vivo. Rac2(-/-) mast cells demonstrated a significant reduction in growth factor-induced survival, which correlated with the lack of activation of Akt and significant changes in the expression of the Bcl-2 family members BAD and Bcl-XL, in spite of a 3-fold induction of Rac1 protein. These results suggest that Rac2 plays a unique role in multiple cellular functions and describe an essential role for Rac2 in growth factor-dependent survival and expression of BAD/Bcl-XL. PMID- 10843389 TI - The Rel protein DIF mediates the antifungal but not the antibacterial host defense in Drosophila. AB - We have isolated two Drosophila lines that carry point mutations in the gene coding for the NF-KB-like factor DIF. Like mutants of the Toll pathway, Dif mutant flies are susceptible to fungal but not to bacterial infections. Genetic epistasis experiments demonstrate that Dif mediates the Toll-dependent control of the inducibility of the antifungal peptide gene Drosomycin. Strikingly, DIF alone is required for the antifungal response in adults, but is redundant in larvae with Dorsal, another Rel family member. In Drosophila, Dif appears to be dedicated to the antifungal defense elicited by fungi and gram-positive bacteria. We discuss in this light the possibility that NF-KB1/p50 might be required more specifically in the innate immune response against gram-positive bacteria in mammals. PMID- 10843390 TI - Caspase-1 activation of IL-1beta and IL-18 are essential for Shigella flexneri induced inflammation. AB - Caspases are intracellular proteases that mediate mammalian cell apoptosis. Caspase-1 (Casp-1) is a unique caspase because it activates the proinflammatory cytokines interleukin (IL)-1beta and IL-18. Shigella flexneri, the etiological agent of bacillary dysentery, induces macrophage apoptosis, which requires Casp-1 and results in the release of mature IL-1beta and IL-18. Here we show that casp 1(-/-) mice infected with S. flexneri do not develop the acute inflammation characteristic of shigellosis and are unable to resolve the bacterial infection. Using casp-1(-/-) mice supplemented with recombinant cytokines and experiments with IL-1beta(-/-) and IL-18(-/-) mice, we show that IL-1beta and IL-18 are both required to mediate inflammation in S. flexneri infections. Together, these data demonstrate the importance of Casp-1 in acute inflammation and show the different roles of its substrates, IL-1beta and IL-18, in this response. PMID- 10843391 TI - Can adhesions be prevented? PMID- 10843392 TI - Changing patterns of severe craniomaxillofacial trauma in Auckland over eight years. AB - BACKGROUND: This study was performed to review the changing pattern of incidence of severe craniomaxillofacial (CMF) trauma in Auckland over 8 years (1989-1997) and to audit the involvement of the regional plastic surgery service. METHODS: A review of prospectively collected admission data of patients admitted to the Auckland Hospital Department of Critical Care Medicine (DCCM) with severe CMF trauma during 1997. A comparison is made with similar data from 1989. Injury severity was defined using the Injury Severity Score (ISS). RESULTS: Twenty-six patients with severe CMF trauma were admitted to Auckland Hospital DCCM in 1997. Their average ISS was 35. Eighty per cent had a significant head injury. Sixty two per cent had injuries due to road traffic accidents (RTA) and 42% had positive blood alcohol levels, including 37% of the RTA victims. Twenty-three per cent had their surgical care provided by the regional plastic surgery service. In 1989, 55 patients were admitted to DCCM with severe CMF trauma. The average ISS was 36. Ninety-five per cent had a significant head injury. Seventy-three per cent had injuries due to RTA and 55% had positive blood alcohol, including 60% of the RTA group. CONCLUSIONS: Patients with severe CMF trauma make up a significant proportion of trauma admissions to DCCM and have a high incidence of life threatening injuries. A multidisciplinary approach is essential. The nature and severity of these injuries has not changed over the last decade. There has been a clear decrease in the incidence of these injuries. This seems to be due to a profound decrease in the rate of RTA associated with alcohol intoxication. PMID- 10843393 TI - Complications of trans-sphenoidal surgery: the Wellington experience. AB - BACKGROUND: All patients who underwent trans-sphenoidal surgery between January 1984 and December 1998 were reviewed to assess morbidity resulting from this operation. METHODS: There were 185 operations on 165 patients. The operative approach was sublabial in 80 cases and transnasal in 105. One surgeon (VB) performed the vast majority of operations. RESULTS: Complications included nasal perforation (7.6%), transient diabetes insipidus (4.9%), permanent diabetes insipidus (3.8%), cerebrospinal fluid fistula (4.3%), donor site haematoma (2.2%) and residual tumour haemorrhage (1.6%) causing ophthalmoplegia (1.1%) and loss of vision (1.1%). Other complications included epistaxis (1.1%), meningitis (0.5%) and sinusitis (0.5%). Injury to the anterior superior alveolar nerve also occurred in the sublabial approach in 6.3% of patients. There were no perioperative deaths. CONCLUSIONS: There is a small but significant risk of a number of complications that should be considered for informed consent of this procedure. PMID- 10843394 TI - Single-versus multiple-dose antibiotics prophylaxis for cardiac surgery. AB - BACKGROUND: This study was carried out to determine if single-dose antimicrobial prophylaxis is sufficient for cardiac surgery. METHODS: The study was a prospective non-randomized trial of 353 consecutive patients undergoing cardiac surgery. Group A (n = 151) received 48 h of prophylaxis and Group B (n = 202) received a single dose. Cephazolin was used in all patients except those at high risk from methicillin-resistant Staphylococcus aureus (MRSA) who received teicoplanin and timentin. RESULTS: There was an overall in-hospital infection rate of 2.8%. There was no significant difference in rate or type of infection between the two groups. CONCLUSIONS: An in-hospital infection rate of 2.8% compares favourably with other reported series. Single-dose antimicrobial prophylaxis is as effective as a 48-h regimen. Targeting high-risk groups is effective. PMID- 10843395 TI - Prevalence and aetiology of lymphoedema after breast cancer treatment in southern Tasmania. AB - BACKGROUND: Lymphoedema can be a devastating complication of surgical treatment for breast cancer. There is a lack of research on its prevalence in Australia which has hindered the development of measures to combat the condition. The aims of this study were to establish the prevalence and investigate the aetiology of upper limb lymphoedema in women treated for breast cancer in the years 1994-1996 in southern Tasmania. METHODS: A standard volumetric water displacement technique was used to measure the arms of 201 women. A subjective assessment of swelling was also made by each patient. Factors analysed for statistical association with lymphoedema were: patient characteristics, type of treatment and tumour, and lymph node pathology. RESULTS: The overall objective prevalence rate, regardless of treatment type, was 11%; whereas, the subjective rate was 23.4%. The objective prevalence for procedures involving axillary surgery was 14.2%. Significant statistical associations were found between arm size and body mass index at time of assessment (r = 0.15, P = 0.04); type of surgery (Chi-squared test = 11.06, P = 0.05); surgery to axilla (U = 2515.5, P = 0.002); tumour size (r = 0.17, P = 0.03); and tumour grade (Chi-squared test = 6.5 1, P = 0.04). No significant relationship was found between lymphoedema and axillary irradiation, number of lymph nodes removed, age or handedness of the patient. CONCLUSIONS: Women receiving axillary dissection as part of their breast cancer treatment carry a significant risk of developing lymphoedema, regardless of the extent of surgery. The causative role of axillary irradiation was not supported. Future research should concentrate on less invasive alternatives to axillary dissection, such as sentinal lymph node biopsy. PMID- 10843396 TI - Axillary dissection and ductal carcinoma in situ of the breast: a change in practice. AB - BACKGROUND: Axillary dissection may be associated with significant morbidity and, while it is necessary in the treatment of invasive breast cancer, is not indicated for the treatment of pure ductal carcinoma in situ (DCIS), although it is being performed in a significant number of cases. The present study examined the incidence of elective axillary dissection in the treatment of DCIS cases detected in a mammographic screening programme over a 4-year period, and whether surgeons have changed their practice in this respect. METHODS: BreastScreen Victoria records were examined retrospectively for the period from January 1995 to December 1998 to identify patients treated for DCIS. The incidence and indications for axillary surgery were investigated. RESULTS: There were 579 cases of DCIS and 93 (16%) had some form of axillary surgery, which was thought to be inappropriate in 57 (10%), the latter being performed by 21 city surgeons and 20 rural surgeons. Before surgery, 36 (63%) cases were diagnosed by core biopsy or excision, and 21 (37%) had imaging and cytology alone for diagnosis. The rate of unnecessary axillary dissections dropped steadily from 14% in 1995 to 4% in 1998, a significant reduction (P = 0.01). CONCLUSION: The incidence of axillary dissection for DCIS has dropped significantly over the last 4 years in Victoria, possibly due to increased awareness through education and guidelines. Surgeons are now more aware that in situ lesions do not need axillary dissection, and that axillary dissection should not be performed for breast cancer unless invasion has been proved histologically. PMID- 10843397 TI - Audit of outpatient laparoscopic cholecystectomy. Universities of Melbourne HPB Group. AB - BACKGROUND: Because the postoperative stay after laparoscopic cholecystectomy (LC) has shortened, it seemed that outpatient LC would be feasible. The aim of this study was to prospectively audit initial experience with outpatient LC at the Austin and Repatriation Medical Centre. We aimed to determine appropriate patient selection criteria, to devise anaesthetic and discharge protocols and to assess patient satisfaction at follow up. METHODS: All patients presenting for LC were assessed for suitability, and those elective cases unlikely to have a duct stone and fulfilling the social criteria were studied. After standard anaesthetic and LC technique, patients recovered in the day surgery unit for up to 8 h and were discharged if stable. The hospital in the home nursing service monitored patients for 48 h and arranged readmission if needed. Patient satisfaction was assessed by independent telephone questionnaire 6 weeks postoperatively. RESULTS: Forty-five patients (median age 43 years) underwent outpatient LC with a discharge rate of 82.3%, resulting in a cost saving of $984 per patient treated. One patient was readmitted, giving an overall success rate of 80%. After stricter implementation of the protocol in the second half of the study, the discharge rate rose to 92%. Patient acceptance of the technique was high at 84.5%. CONCLUSIONS: The results of the first 45 patients show that it is possible to safely perform outpatient LC with a low admission rate in fit, elective patients who live close to medical care. Provided a strict anaesthetic protocol is followed, the technique has good patient acceptance and provides some economic benefit to the hospital. PMID- 10843398 TI - Gangrenous cholecystitis in the laparoscopic era. AB - BACKGROUND: We reviewed our data of laparoscopic cholecystectomies between 1990 and 1997 with reference to gangrenous cholecystitis. METHODS: In a consecutive series of 1304 patients having laparoscopic cholecystectomies, prospective data collection has permitted analysis of the relationship between gangrenous cholecystitis (GC), acute (non-gangrenous) cholecystitis (AC) and non-acute cholecystectomies (NAC). RESULTS: Twenty-five patients had gangrenous cholecystitis and 238 had acute cholecystitis. We found that patients with GC were significantly older (65.4 years vs 56.1 years (AC) and 52.7 years (NAC), P < 0.05) and had a higher M: F ratio (1.5:1 vs 1:2.6 (AC) and 1:2.8 (NAC), P < 0.05). Cardiac disease was found to be a significant factor but not diabetes. Preoperative ultrasonography correctly identified only 17 patients with acute inflammatory changes. Seven patients had an absent sonographic Murphy's sign. The gall bladder wall was generally thicker (4.11 mm vs 3.8 mm (AC) and 2.7 mm (NAC), P < 0.05) but there was marked overlap between the three groups. The common bile duct (CBD) was more dilated (6.1 mm vs 4.8 mm (AC) and 4.6 mm (NAC), P < 0.006) and there was increased incidence of CBD stones in the GC group. Our conversion rate was 8.7% with minimal complications and no operative mortality. CONCLUSION: Patients with GC were generally older, more likely to be male and had increased incidence of cardiovascular disease. Preoperative ultrasound cannot accurately identify those patients with gangrenous cholecystitis, but with conversion rates of 8.7% and no operative mortality, they can generally be managed safely with laparoscopic surgical techniques. PMID- 10843399 TI - Joint-preserving operation for osteoarthrosis of the hip in adult cerebral palsy. AB - BACKGROUND: A joint-preserving operation was performed on 15 hips with osteoarthrosis, involving 12 patients who had adult cerebral palsy. METHODS: Eleven hips underwent Chiari pelvic osteotomy only; three hips underwent Chiari pelvic osteotomy with femoral osteotomy and the other one hip underwent femoral varus osteotomy only. The mean follow-up period after surgery was 6 years and 2 months (with follow-up range of 2 years and 3 months to 10 years and 6 months). RESULTS: Good results were achieved in 13 of the 15 hips (86.6%). Two patients with athetotic tetraplegia treated with Chiari pelvic osteotomy had pelvic obliquity. Progressive osteoarthrotic change continued in bilateral hips in one case treated with Chiari pelvic osteotomy. CONCLUSION: We confirm that usual treatment for osteoarthrosis of the hip was also applicable for osteoarthrosis of the hip in cases of adult cerebral palsy, provided sufficient attention is given to the complications accompanying spastic paralysis. PMID- 10843400 TI - Duplicated popliteal and superficial femoral veins: incidence and potential significance. AB - BACKGROUND: Duplication of the popliteal and superficial femoral veins (PV, SFV) is a normal variant previously reported in up to 25% of limbs. Little clinical significance, however, has been attributed to this apparently common anomaly. The present study was designed to determine the incidence of duplications in individuals presenting for venous incompetence studies, and whether their presence could, in theory, act as a predisposing factor to deep venous thrombosis (DVT) formation. METHODS: Duplex ultrasound examinations were performed in which venous duplications were actively searched for and recorded. The diameters of both limbs of any duplicated system and the single vessel immediately distal to it were recorded. Using these measurements, the changes in total cross-sectional area (CSA) associated with these anomalies were calculated. In addition, with the knowledge that the volume flow rate must remain constant, the velocity changes associated with such systems were calculated. RESULTS: A total of 248 limbs from 177 patients was scanned. Duplications were found in 39 (15.7%) of these limbs. Of these, 30 limbs (77%) involved only the SFV, seven (18%) involved both the SFV and PV, and two (5%) involved only the PV. Short-segment SFV duplications were used to calculate the percentage change in total CSA and therefore blood flow velocities. Of the 13 (33%) suitable for such calculations, and calculating for each individual duplicated system, a mean increase in the vessel's total CSA of 42%, which corresponded to a theoretical decrease in blood flow velocity of 36%, was found. CONCLUSION: The present study confirms the significantly high incidence of duplications of the PV and SFV and raises the possibility of the potential for DVT formation secondary to changes in flow velocities. PMID- 10843401 TI - Testicular torsion: time is the enemy. AB - BACKGROUND: The acute scrotum is a diagnostic dilemma, and testicular torsion is of primary interest because of its fertility problems for the patient and medico legal issues for the surgeon. The present study aimed to correlate operative findings of patients with suspected testicular torsion with certain clinical variables and investigations to see if diagnosis and outcome could be improved. METHODS: A total of 99 patients underwent scrotal exploration for suspected testicular torsion at the Royal Brisbane Hospital between 1990 and 1995. Colour Doppler ultrasound, white blood count and urine microscopy results were documented, along with the patient's age and duration of testicular pain. RESULTS: Fifty-six patients were found to have torsion, and the testicular loss rate was 23%. Patients who experienced testicular pain for longer than 12 h had a testicular loss rate of 67%. A negative urine microscopy was suggestive of testicular torsion, but was not diagnostic. The white blood count did not aid in the diagnosis. Colour Doppler ultrasound of the scrotum was used on nine occasions with three false negative results and a sensitivity of only 57%. CONCLUSIONS: Time is the enemy when managing the acute scrotum. No investigation substantially improves clinical diagnosis enough to warrant any delay in definitive surgical intervention. PMID- 10843403 TI - Early experience with clinical indicators in surgery. AB - BACKGROUND: In 1997 a set of 53 clinical indicators developed by the Royal Australasian College of Surgeons (RACS) and the Australian Council on Healthcare Standards (ACHS) Care Evaluation Programme (CEP), was introduced into the ACHS Evaluation and Quality Improvement Programme (EQuIP). The clinical indicators covered 20 different conditions or procedures for eight specialty groups and were designed to act as flags to possible problems in surgical care. METHODS: The development process took several years and included a literature review, field testing, and revision of the indicators prior to approval by the College council. In their first year 155 health-care organizations (HCO) addressed the indicators and this rose to 210 in 1998. Data were received from all states and both public and private facilities. RESULTS: The collected data for 1997 and 1998 for some of the indicators revealed rates which were comparable with those reported in the international literature. For example, the rates of bile duct injury in laparoscopic cholecystectomy were 0.7 and 0.53%, respectively; the mortality rates for coronary artery graft surgery were 2.5 and 2.1%, respectively; the mortality rates after elective abdominal aortic aneurysm repair were 2.5 and 3.7%, respectively; and the post-tonsillectomy reactionary haemorrhage rates were 0.9 and 1.3%, respectively. Results for some indicators differed appreciably from other reports, flagging the need for further investigation; for example, the negative histology rates for appendectomy in children were 18.6 and 21.2%, respectively, and the rates for completeness of excision of malignant skin tumours were 90.7 and 90%, respectively. The significance of these figures, however, depends upon validation of the data and their reliability and reproducibility. Because reliability can be finally determined only at the hospital level they are of limited value for broader comparison. CONCLUSION: The process of review established for the indicator set has led to refinement of some indicators through improvement of definitions, and to a considerable reduction in the number of indicators to 29 (covering 18 procedures), for the second version of the indicators (which was introduced for use from January 1999). The clinical indicator programme, as it has with other disciplines, hopefully will provide a stimulus to the modification and improvement of surgical practice. Clinician ownership should enhance the collection of reliable data and hence their usefulness. PMID- 10843402 TI - Surgical adhesions: evidence for adsorption of surfactant to peritoneal mesothelium. AB - BACKGROUND: It has been speculated that the formation of surgical adhesions must be preceded by physical adhesion of the two surfaces, a process normally prevented by a lining of adsorbed surface-active phospholipid (surfactant) acting as both a superb boundary (solid-to-solid) lubricant and a release (antistick) agent. Animal trials administering exogenous surfactant as a dry powder (ALEC) have previously demonstrated a reduction of 80% in abdominal adhesions. METHODS: Incubation of rat peritoneum (both live and excised) with radiolabelled dipalmitoyl phosphatidylcholine (DPPC) has been used to demonstrate adsorption; while the normal lining of surfactant in the human abdominal cavity has been confirmed by epifluorescence microscopy using Phosphin E as the hydrophobic probe. AIMS: The overall aim is to confirm that peritoneal mesothelium has a lining of surfactant known for its lubricating and release properties, and that this lining can be enhanced by the adsorption of exogenous material. RESULTS: Adsorption of DPPC to peritoneal mesothelium was 470 ng/cm2 (n = 8) ex vivo and 598 ng/cm2 (n = 18) in vivo, these rates being enhanced by EggPG by 62% ex vivo and 47% in vivo to reach the equivalent of almost three close-packed monolayers. CONCLUSIONS: These results can explain the reduction in surgical adhesions previously reported in animals by administering ALEC (7:3 DPPC:EggPG) as a highly surface-active dry powder, although it is now used in saline suspension to treat respiratory distress syndrome in newborns, in whom it has no side-effects. These findings would appear to justify clinical trials for dry ALEC in suppressing surgical adhesions with minimal risk of an adverse reaction. The results of these trials are also discussed and found to be compatible with the known ability of surfactant to resist physical adhesion by fibronectin, the tacky 'glue' by which fibroblasts attach to surfaces as the first step in formation of fibrinous adhesions. PMID- 10843404 TI - Colorectal cancer chemoprevention: aspirin, other NSAID and COX-2 inhibitors. PMID- 10843405 TI - Surgical training of doctors in their own isolated hospital. PMID- 10843406 TI - Popliteal artery compression by a tibial osteochondroma with associated thrombosis and distal embolization. PMID- 10843407 TI - Intramuscular haemangioma of the medial pterygoid. PMID- 10843408 TI - Retroperitoneal textiloma following laparoscopic-assisted nephro-ureterectomy for lower ureteric cancer, masquerading as a metastatic soft-tissue tumour. PMID- 10843409 TI - Embolization as a treatment for bleeding post-total hip arthroplasty. PMID- 10843410 TI - Pig endogenous retrovirus--a threat to clinical xenotransplantation? AB - Transplantation shows good results for patients with end-stage disease, but there is an increasing lack of organs. Xenotransplantation, the transfer of live animal cells, tissues, or organs to another species, offers a potential solution to this shortfall. Pig is regarded as the animal of choice for this purpose. Meanwhile demonstration of pig endogenous retrovirus (PERV) in all porcine herds has caused serious concern with respect to a possible transmission of the virus to humans with a transplanted organ. Transmission to human cells has been documented under certain in vitro conditions. However, no such transmission has been demonstrated in vivo. The possible consequences of introducing PERV into immunocompromised human organisms are not known and it is necessary to collect more information. Novel and sensitive genomic assays to detect PERV infection are now available in addition to established virological, immunoserological and molecular methods. In order to minimise the risk of PERV transmission rigorous procedures should be established. International guidelines to reduce the risk should be followed. Although a number of immunological, physiological and virological questions need to be answered before the introduction of xenotransplantation as an alternative clinical treatment, some problems can only be solved by judicious clinical trials. PMID- 10843411 TI - Disease-dependent changes in T-cell populations in patients with shigellosis. AB - In shigellosis, bacterial infection is associated with an extensive inflammation of the rectal mucosa, resulting in bloody dysentery. The role of T-cell-mediated pro-inflammatory mechanisms has been implicated in this process, but the specific role of T-cell subsets is still not well understood. In this study we attempted to identify the changes in T-cell populations in patients with shigellosis during the disease course. The T-cell subset distribution was analyzed by immunohistochemistry in the rectal mucosa and by immuno-flow cytometry in the peripheral blood. Blood and rectal biopsies were studied from patients with Shigella dysenteriae 1 (n= 11) and S. flexneri (n= 11) infection and 20 healthy age-matched controls. We found an expansion of gammadelta+T cells in the rectal mucosa, but a decrease in the percentage of gammadelta+T cells in the blood in acute shigellosis. There was also a preferential increase in CD8+ T cells in the surface epithelium of rectal tissue in patients infected with S. dysenteriae 1, but not in patients infected with S. flexneri. Our findings suggest that the rectal mucosal inflammation in shigellosis is associated with an expansion of T cells, in particular CD8+ and gammadelta+T-cell subsets in the gut mucosa, which may be of importance for the pathogenesis of shigellosis. PMID- 10843412 TI - Evaluation of post-antibiotic effects of antipseudomonal antibiotics using an automated system. AB - Different methods have been described for determination of the post-antibiotic effects (PAEs) of antibiotics, from the conventional methods to the automatic measurement of bacterial regrowth. The PAEs of ciprofloxacin, ofloxacin and perfloxacin, as compared with those of amikacin, netilmicin and ceftazidime, have now been investigated for three clinical isolates of Pseudomonas species, using automated measurement of the growth with an Anthos microplate reader. It was found that, besides the well-known PAEs of aminoglycosides, quinolone antibiotics also exhibit drug- and concentration-dependent PAEs against clinical isolates of Pseudomonas when used in concentrations of 1/2x, 1 x or 5 x MIC. Of the three quinolones tested, pefloxacin showed the greatest PAE, independently of its MICs against the different strains. Ofloxacin had no or only an insignificant PAE, while ciprofloxacin had a marked PAE for all strains, including the reference strains. PMID- 10843413 TI - Sequence analysis of the hemagglutinin gene of measles virus isolates in Denmark 1997-1998: no evidence of persistent circulation of measles virus in Denmark. AB - The hemagglutinin-coding region of 17 virus samples from 12 measles cases in Denmark during 1997-1998 was analysed by partial nucleotide sequencing. The cases appeared as three sporadic cases and two epidemics, both with a limited time course and geographical distribution. The measles strains identified from the three sporadic cases and two epidemics could be allocated to five different previously well-defined sequence groups consistent with the assumption that cases of measles in Denmark are due to repeated introduction from abroad rather than persistent circulation of strains in the population. PMID- 10843414 TI - Histopathological evidence of North American blastomycosis in Italy: report of two cases. AB - No clinical reports of blastomycosis in Italy have been published until now. We here report two cases of histologically diagnosed, unexpected cutaneous involvement in patients, aged 78 and 52 years, living in North Italy and never having been abroad. The histological differential diagnosis between blastomycosis and other fungal pathogens is discussed. Even in the absence of culture the present cases can confidently be considered as genuine examples of Blastomyces dermatitidis infection in Italy. PMID- 10843415 TI - The effects of live Neisseria meningitidis and tumour necrosis factor-alpha on neutrophil oxidative burst and beta2-integrin expression. AB - The effects of human recombinant tumour necrosis factor-alpha (TNF-alpha) on neutrophil (PMNL) oxidative burst and on CD11b/CD18 and CD14 expression after stimulation with pathogenic or nonpathogenic Neisseria meningitidis were studied using chemiluminescence and flow cytometry. PMNL oxidative burst increased more when stimulated with the apathogenic 29E strain than with the pathogenic B strain both when studied by chemiluminescence and by flow cytometry. When TNF-alpha was added to whole blood or PMNL together with bacteria a significant increase in the oxidative burst was seen for the B strain only. When whole blood was preincubated for 30 min with TNF-alpha the increase in oxidative burst was significant for both meningococcal strains. TNF-alpha caused a significant increase in PMNL CD 11b/CD18 expression after 30 min of incubation at 37 degrees C. TNF-alpha added simultaneously with the bacteria induced a significant increase in PMNL CD11b/CD18 in both strains. Incubation with the B strain alone caused a low but significant increase in CD11b/CD18 expression, but the addition of TNF-alpha increased this expression to the same high level as incubation with TNF-alpha alone or the 29E strain alone. Only TNF-alpha and the 29E strain caused significant increases in CD14 expression. In conclusion, human PMNLs react differentially when stimulated with pathogenic and nonpathogenic N. meningitidis and the activating effect of TNF-alpha is variable depending on the bacteria involved. PMID- 10843416 TI - Vasitis nodosa: immunohistochemical findings--case report. AB - We report the immunohistochemical features of vasitis nodosa and discuss the differential diagnosis. The patient was a 42-year-old Japanese man with bilateral small indurations of the vas deferens at the site of a previous vasectomy. Microscopically, small-sized ducts proliferated within the muscular wall of the vas deferens, and focally in the surrounding connective tissue. Immunohistochemically, most proliferating glandular cells were strongly positive for cytokeratins 7, 19, and 34betaE12, and vimentin. Epithelial membrane antigen and Leu-M1 reacted against the luminal surface of the cells. Focally, glandular cells were also positive for CA125. Cytokeratin 20, carcinoembryonic antigen, and prostate-specific antigen were negative. We discuss the immunohistochemical differentiation of vasitis nodosa from prostatic adenocarcinoma, adenocarcinoma of the rete testis, and adenomatoid tumor. PMID- 10843417 TI - Intravenous tufted angioma. AB - A case of a rare vascular tumor, intravenous tufted angioma, is described. A 51 year-old Japanese man presented with a 12x8 mm solitary reddish nodule on the right foot, which had been found at birth. Histologically, the tumor was confined to a malformed vein and was characterized by nodular aggregates of plump cells. The aggregates showed a compact proliferation of round cells, including capillary forming cells. Venous angiomatous areas were also observed. No multinucleated giant cells were seen. Immunohistochemically, the capillary-forming cells in the aggregates and the endothelial cells in the angiomatous areas were positive for endothelial markers (factor VIII-related antigen, CD31, CD34). Pericyte-like cells expressing alpha-smooth muscle actin and muscle actin, and macrophage-like cells, which stained for factor XIIIa, were intermingled in the cellular aggregates. Flow cytometric analysis showed diploidy. The tumor may be a hamartomatous lesion modified by secondary reactive changes, and it may represent a distinctive clinicopathological entity that is closely related histogenetically and perhaps pathologically to tufted angioma and the recently described "giant cell angioblastoma". PMID- 10843418 TI - Vacuolating cytotoxic activity of 40 Helicobacter pylori strains isolated from Turkish patients. AB - In this study we examined the in vitro vacuolating cytotoxic activity of Helicobacter pylori, which is a gram-negative microaerophilic curved bacterium and a causative agent of gastritis, peptic ulcer and gastric ulcer. A vacuolating cytotoxin assay was performed to assess the vacuolating activity of 40 strains (20 gastritis, 11 gastric ulcer, and 9 duodenal ulcer), which were obtained from patients undergoing upper gastrointestinal endoscopy. The Vero cell line was used in the cytotoxic assay. Of the 40 isolates, 24 (12 gastritis, 6 gastric ulcer, 6 duodenal ulcer) were cytotoxic for the Vero cell line at 1:4 and 1:8 dilutions. Thus, vacuolating cytotoxin of H. pylori affects the Vero cell line, but it seems there is no correlation between the positivity of the strains and the risk of any particular H. pylori disease. PMID- 10843419 TI - Outbreak of ampicillin-resistant Enterococcus faecium--risk factors for faecal colonisation. AB - Since January 1995 there has been a nosocomial outbreak at Haukeland University Hospital involving more than 330 patients with clinical infections caused by ampicillin-resistant Enterococcus faecium (ARE) (minimum inhibitory concentration > or =32 mg/l). Rectal carriage of ARE was initially observed on two medical wards only. Here the ARE colonisation rate has remained high. To assess risk factors for ARE colonisation we performed a case-control study including 37 rectal carriers of ARE and 83 non-carriers on these wards. Significant differences were found between cases and controls with respect to the mean number of days on antimicrobial treatment (13.3 for carriers, 5.5 for non-carriers, p<0.001), mean number of different antibiotics prescribed (2.8 for carriers, 2.1 for non-carriers, p= 0.008) and mean number of days in hospital (18.4 vs 10.2, p=0.001). Unadjusted statistical analysis showed that several antibiotics were risk factors for ARE carriage. Logistic regression analysis showed that fluoroquinolone prescription (OR=3.5, p=0.01) and more than 10 days of antibiotic use (OR= 3.3, p=0.01) were significant risk factors. An additional follow-up screening of previous carriers revealed no colonisation 8 to 36 (median 9) months after discharge from hospital (n=17). Prolonged antimicrobial therapy and broad spectrum antibiotics seem to facilitate nosocomial ARE colonisation. PMID- 10843420 TI - Emm gene polymorphism among temporally clustered group A streptococcal isolates in western Norway. AB - Nineteen group A streptococcal isolates obtained in western Norway from patients with invasive disease during a period of high morbidity and mortality were examined for clonality and emm gene polymorphism. These isolates belonged to the prevalent serotypes during the outbreak, namely T1, T3 or T6. Restriction fragment length polymorphism and sequencing of the emm genes were used to compare these isolates with 14 isolates of the same serotype but from non-invasive infections. The restriction analysis did not identify specific invasive clones. The emm genes in three of the four T3 isolates from invasive disease had nucleotide substitutions inducing a charge difference in the N-terminal part of the M protein. The 4 T6 isolates had a longer emm amplicon when compared to 15 isolates from superficial infections and also showed nucleotide substitutions that could induce conformational changes in the hypervariable end of the M protein. Restriction analysis of the emm amplicon of the T6 isolates in order to estimate the number of A- and C-repeats is described. The emm gene sequence served as an epidemiological marker within the serotypes T3 and T6, but the significance of the emm polymorphism displayed by the isolates from invasive disease is uncertain at this stage. PMID- 10843421 TI - Leukotrienes, leukotriene receptor antagonists, and rhinitis. PMID- 10843423 TI - Evidence of eosinophil, neutrophil, and mast-cell mediators in the effusion of OME patients with and without atopy. AB - BACKGROUND: This study was designed to measure the involvement of eosinophils, neutrophils, and mast cells in the middle ear of patients with OME and to see whether that inflammatory response differed depending on whether or not the patient was atopic. METHODS: Levels of ECP, MPO, and tryptase were measured in effusion from 97 patients whose atopic status was determined by in vitro testing with 12 inhalants and five foods. RESULTS: Seventy-nine of 97 OME patients (81%) were atopic, among whom 86% (68/79) had elevated levels of effusion ECP, and 64% (23/36) had elevated tryptase. Mean ECP was 166 microg/l vs 3.38 microg/l, mean MPO was 6231 microg/l vs 116 microg/l, and mean tryptase was 4.8 microg/l vs 1.34 microg/l in atopic vs nonatopic ears. Total serum IgE did not differ between atopics and nonatopics (P=0.28). CONCLUSIONS: The inflammatory response by eosinophils, neutrophils, and mast cells in the middle ear was distinctly different between atopic and nonatopic patients (P < 0.001). This study provides evidence that eosinophils and mast cells, both essential to a Th-2 driven immune response, are present in the majority of ears with chronic OME, and supports the hypothesis that middle-ear mucosa, like that of the rest of the upper respiratory tract, is capable of an allergic response. PMID- 10843422 TI - The potential of active metabolites of antihistamines in the management of allergic disease. PMID- 10843426 TI - Early effect of ultrarush venom immunotherapy on the IgG antibody response. AB - BACKGROUND: We have previously shown in several allergy models that allergic and tolerance status with respect to allergens is associated with a somewhat different dominant specificity of IgG antibodies. The objective was to test this hypothesis in the compelling model of ultrarush venom immunotherapy (VIT), which induces clinical tolerance after only a few hours of treatment. METHODS: Antibody titers and specificity were evaluated through solid-phase ELISA using streptavidin-biotin technology in 12 patients allergic to wasp venom before and during the ultrarush procedure (at 12 h, 24 h, and 15 days). The results were compared with those from another group of 20 patients treated with venom injections for at least 2 years. RESULTS: No significant change was observed in IgG titers during the early phase of VIT. The capacity of individual sera to prevent the antigen binding of pooled IgG from allergic patients changed rapidly, with mean percentage inhibitions falling from 80+/-15%, before starting VIT, to 26+/-14%, 35+/-15%, and 34+/-5% after 12 h, 24 h, and 15 days of treatment, respectively (P<0.001 by one-way ANOVA). The capacity of individual sera to prevent the antigen binding of pooled IgG from patients receiving prolonged VIT changed, with mean percent inhibitions increasing from 47+/-8%, before starting VIT, to 76+/-7%, 83+/-6%, and 87+/-6% after 12 h, 24 h, and 15 days of treatment, respectively (P<0.001 by one-way ANOVA). CONCLUSIONS: During the initial phase of ultrarush VIT, a change in IgG specificity, i.e., a change in the set of epitopes dominantly recognized by IgG on wasp-venom antigens, occurred concomitantly with early clinical tolerance and was already detectable a few hours after the onset of treatment. Although it may be an epiphenomenon, this change represents the earliest humoral modification described so far during this procedure. The mechanism is unknown, but it appears to be a selective depletion of the highest avidity antibody fraction by the venom injected in large doses at this stage of therapy. Finally, our data now show the previously documented association between a particular IgG specificity and the clinical status (allergy vs tolerance) to be true also with ultrarush VIT, a model in which the clinical ability to display allergic symptoms is rapidly reversed. PMID- 10843425 TI - Dendritic cell number is related to IL-4 expression in the airways of atopic asthmatic subjects. AB - BACKGROUND: Airway dendritic cells are essential for stimulating naive T cells in response to inhaled antigen and for the development of allergic sensitization. IL 4 in vitro can distinguish dendritic cell lines from peripheral blood mononuclear cells. Our study had the following aims: 1) to compare the distribution of CD1a+ dendritic cells and IL-4+ cells, in the bronchial mucosa of asthmatics and controls 2) to determine the relationship between the numbers of CD1a+ dendritic cells and IL-4+ cells in the bronchial mucosa of asthmatics 3) to determine whether CD1a+ cells express the IL-4 receptor. METHODS: Twenty atopic asthmatic and eight normal subjects were studied. In each subject, bronchoscopy with bronchial biopsies was performed. CD1a, IL-4, and IL-4 receptor expressions were evaluated by immunohistochemistry. RESULTS: The number of CD1a+ and IL-4+ cells was significantly higher in asthmatics than controls. The number of CD1a+ cells was positively correlated to the number of IL-4 + cells. Bronchial biopsy serial section studies showed that CD1a+ cells express the receptor for IL-4. CONCLUSIONS: These results suggest that an increased amount of IL-4 may play a physiopathologic role in maintaining the dendritic cell pool in vivo. Therefore, because of possible IL-4 activity on antigen-presenting cells in T-cell immune responses to allergens, an important new role of IL-4 in asthma inflammation can be envisaged. PMID- 10843427 TI - Regulation by intracellular glutathione of TNF-alpha-induced p38 MAP kinase activation and RANTES production by human pulmonary vascular endothelial cells. AB - BACKGROUND: We have previously shown that p38 mitogen-activated protein (MAP) kinase regulates tumor necrosis factor-alpha (TNF-alpha)-induced RANTES production by human pulmonary vascular endothelial cells, and that sensitivity to TNF-alpha is inversely correlated with cellular reduction and oxidation (redox) state. However, a regulatory role of intracellular glutathione (GSH) in TNF-alpha induced p38 MAP kinase activation and p38 MAP kinase-mediated RANTES production has not been determined. In the present study, therefore, we extended our previous studies and focused on redox regulation on p38 MAP kinase activation. METHODS: Human pulmonary vascular endothelial cells were exposed to N acetylcysteine (NAC) or buthionine sulfoximine (BSO), and then TNF-alpha-induced p38 MAP kinase activation and p38 MAP kinase-mediated RANTES production were determined. RESULTS: The results showed that 1) NAC attenuated TNF-alpha-induced p38MAP kinase activation and RANTES production 2) SB 203580 as the specific inhibitor of p38 MAP kinase activity attenuated TNF-alpha-induced RANTES production 3) BSO facilitated TNF-alpha-induced p38 MAP kinase activation and RANTES production 4) SB 203580 attenuated BSO-mediated facilitation of TNF-alpha induced RANTES production. CONCLUSIONS: These results indicated that TNF-alpha induced p38 MAP kinase activation and p38 MAP kinase-mediated RANTES production by human pulmonary vascular endothelial cells are inversely regulated by intracellular GSH levels. PMID- 10843428 TI - Effects of maternal allergen-specific IgG in cord blood on early postnatal development of allergen-specific T-cell immunity. AB - BACKGROUND: A wide body of epidemiologic evidence indicates that as yet unknown maternal factor(s) can influence susceptibility to allergic disease in the offspring. It is also well established that the induction of allergen-specific T cell memory is frequently initiated in utero, and it is likely that maternal factors exert their influence during this period. METHODS: This study examines the relationship between maternally derived allergen-specific IgG subclass antibodies and cellular immune responses (lymphoproliferation and cytokine production) against the same allergens in 49 subjects tested at birth and at 2 years of age. Polyclonal production of the Th1 cytokine IFN-gamma was also examined in the cord-blood samples. RESULTS: At birth, there were positive correlations between both house-dust mite (HDM)- and ovalbumin (OVA)-specific IgG subclass levels in cord blood, maternal atopy, and the magnitude of perinatal lymphoproliferative responses to respective allergens. Inverse relationships were also observed between cord-blood IgG antibody titres and allergen-specific production of some Th2 cytokines. However, there were no consistent relationships between cord-blood allergen-specific IgG antibodies and subsequent immune responses to allergens when the same subjects were retested at 2 years of age. An inverse relationship was observed between maternal history of atopy and perinatal IFN-gamma production capacity. CONCLUSIONS: Our results suggest that transplacental transfer of allergen-specific IgG antibody is unlikely to be a major mechanism for maternal regulation of allergen-specific immunity in infancy. An alternative possibility is that maternal effects may operate by influencing IFN-gamma production by T cells in the offspring. PMID- 10843429 TI - Is there a role for immunoblots in the diagnosis of latex allergy? Intermethod comparison of in vitro and in vivo IgE assays in spina bifida patients. AB - BACKGROUND: The best diagnostic method for latex allergy is still controversial. This investigation was designed to evaluate the diagnostic efficiency of immunoblotting in comparison with established in vitro and in vivo test systems. METHODS: A total of 108 spina bifida patients were investigated by questionnaire and skin prick test (SPT). Specific serum IgE to latex was analyzed by the Pharmacia CAP FEIA immunoassay, DPC AlaSTAT microplate immunoassay, and DPC AlaBLOT immunoblot. Patients were regarded as latex allergic if they reacted positively to challenge by the latex glove wearing test. RESULTS: Thirty-four patients reacted positively to challenge. The sensitivity rates were 97% (SPT), 94% (immunoblot, CAP), 74% (AlaSTAT), and 35% (clinical history). The specificity rates were 92% (clinical history), 88% (AlaSTAT), 77% (SPT), 76% (CAP), and 69% (immunoblot). If two methods were combined, efficiency rates were highest for SPT combined with CAP (sensitivity 94%, specificity 82%), with AlaSTAT (sensitivity 74%, specificity 92%), or with immunoblot (sensitivity 91%, specificity 84%). The sera of challenge-positive patients recognized more immunoblot bands than challenge-negative patients, and the severity of symptoms correlated with the number of recognized bands. CONCLUSIONS The diagnostic efficiency of immunoblotting is not superior to that of SPT. However, immunoblotting may serve as an additional tool to increase slightly the specificity of SPT and specific serum IgE tests. PMID- 10843430 TI - Reduction of side-effects from ultrarush immunotherapy with honeybee venom by pretreatment with fexofenadine: a double-blind, placebo-controlled trial. AB - BACKGROUND: Immunotherapy with Hymenoptera venoms is highly effective but causes allergic side-effects frequently, especially when honeybee venom is used. Therefore, our objective was to investigate the effect of pretreatment with the antihistamine fexofenadine on the incidence of allergic side-effects during ultrarush immunotherapy with bee venom. METHODS: In a double-blind, placebo controlled trial, 57 patients with a history of systemic allergic reactions to honeybee stings and positive diagnostic tests (skin tests, serum specific IgE to honeybee venom) were investigated. Bee venom immunotherapy was started with an ultrarush protocol and patients were randomized to pretreatment with either fexofenadine 180 mg or placebo on days 1, 8, 22, and 50 of the protocol. Local and systemic allergic side-effects were registered. RESULTS: Fifty-four patients completed the study, 28 on fexofenadine and 26 on placebo pretreatment. On day 1, large local reactions were significantly reduced in both extension and duration by fexofenadine pretreatment (P<0.025). Systemic allergic side-effects on the whole were not reduced. However, the symptoms pruritus, urticaria, and angioedema occurred less frequently with fexofenadine (P<0.05). CONCLUSIONS: Pretreatment with fexofenadine during venom immunotherapy reduces local allergic reactions and generalized symptoms of the urticaria and angioedema type. PMID- 10843431 TI - Increased interleukin-10 and macrophage inflammatory protein-1 alpha release from blood monocytes ex vivo during late-phase response to allergen in asthma. AB - BACKGROUND: We determined the effect of late-phase responses to allergen challenge of mildly allergic asthmatic patients on the expression and release of the cytokines IL-10 and MIP-1alpha from peripheral blood monocytes. METHODS: Sixteen mildly allergic asthmatics were exposed to increasing concentrations of allergen aerosol. Nine demonstrated an early response alone (single responders), while seven had an early followed by a late response (dual responders). Monocytes were isolated from venous blood before and 24 h after allergen challenge, and placed in short-term culture for determination of IL-10 mRNA expression by reverse-transcription polymerase chain reaction and protein release. MIP-1alpha protein release was measured by radioimmunoassay. RESULTS: IL-10 mRNA expression was significantly higher in dual responders than early responders alone, for unstimulated monocytes or for monocytes exposed to lipopolysaccharide or IL 1beta. In addition, the release of IL-10 and MIP-1alpha from monocytes of dual responders was also enhanced. CONCLUSIONS: Circulating monocytes are primed to release more of the cytokines, IL-10 and MIP-1alpha, in dual but not in single responders, at 24 h after allergen. They may contribute to allergen-induced inflammatory responses in the airways. PMID- 10843432 TI - Mustard allergy in children. AB - BACKGROUND: Mustard allergy is not well known. This study aimed to assess its clinical features and other associated allergies, and to define skin prick tests (SPT), specific IgE, and dose response by oral food challenge. METHODS: Our study investigated 36 children with positive mustard SPT. The diagnosis of mustard allergy was based on open or single-blind, placebo-controlled food challenge (SBPCFC). We compared the subjects to 22 controls. RESULTS: The initial clinical features were atopic dermatitis (51.8%), and urticaria and/or angioedema (37%). Fifteen children were allergic (positive SBPCFC) and 21 children were nonallergic (negative SBPCFC). Symptoms after mustard ingestion started under 3 years of age in 53.3% of the subjects. There was no significant difference in the food allergies and associated inhalant allergen sensitizations between the two groups. In the allergic group, the mean wheal diameter for mustard SPT was 8.8 mm and the median concentration of mustard serum (s) IgE 14.8 kU/l. The mean cumulative reactive dose were 153 mg. CONCLUSIONS: Allergic reactions to mustard started early in life. Clinical symptoms were not severe in children. Mustard should be included in screening tests of food allergy in children. PMID- 10843433 TI - Sensitivity to fungal allergens is a risk factor for life-threatening asthma. AB - BACKGROUND: Previous studies have suggested that sensitivity to Alternaria and Cladosporium may be risk factors for life-threatening asthma. We have investigated this by studying the relationship between skin tests for fungal spores and admission to an intensive care unit (ICU) for asthma. METHODS: Skin prick tests for fungal spores (Alternaria tenuis, Cladosporium cladosporoides, Helminthosporium maydis, and Epicoccum nigrum), cat dander, house-dust mite (Dermatophagoides pteronyssinus), and a seven-grass mix were performed in three groups of patients: patients admitted to an ICU with an attack of asthma; those who had received emergency treatment for asthma but had not been admitted to an ICU, and those who had never required emergency treatment for their asthma. RESULTS: Twenty of 37 patients (54%) admitted to the ICU had a positive skin test for one or more fungal allergens compared with 15/50 patients (30%) in each of the other groups (P=0.005). The ICU patients were no more likely to have positive skin tests for the grass mix, cat dander, or house-dust mite than the other patients. CONCLUSIONS: A positive skin test for fungal allergens is a risk factor for admission to an ICU with an acute attack of asthma. PMID- 10843435 TI - Palmar exfoliative exanthema to amoxicillin. PMID- 10843434 TI - Detection and treatment of early asthma. AB - BACKGROUND: We sought answers to two questions: 1) how can mild, early asthma best be detected? 2) how should it be treated and the treatment effect monitored? METHODS: Eighty adult patients with early, mostly mild asthma and 30 control subjects were examined by lung-function and biochemical tests. Patients were randomly assigned to two treatment groups. One group was treated with an inhaled steroid (budesonide) for 6 weeks, and the other first with an inhaled beta2 agonist (terbutaline) for 6 weeks and then with an inhaled steroid for 2 weeks. RESULTS: Treatment with budesonide was effective: symptom scores, PEF, blood eosinophils, and sputum ECP values all improved. Terbutaline was ineffective by these criteria. For the detection of early asthma and for the following treatment effects, sputum and serum ECP assays are useful supplements to lung-function tests. CONCLUSIONS: Treatment of early, mild asthma with an inhaled steroid is effective and worthwhile. Detection of the disease remains a problem, as both lung-function and biochemical tests have low sensitivities. PMID- 10843436 TI - Anaphylaxis to red currants. PMID- 10843437 TI - Swiss chard-induced asthma. PMID- 10843438 TI - Hypersensitivity to Viburnum rhytidophyllum. PMID- 10843439 TI - Exercise-induced anaphylaxis to snails. PMID- 10843440 TI - Assessing phytoestrogen exposure in epidemiologic studies: development of a database (United States). AB - BACKGROUND AND OBJECTIVE: Phytoestrogens (weak estrogens found in plants or derived from plant precursors by human metabolism) have been hypothesized to reduce the risk of a number of cancers. However, epidemiologic studies addressing this issue are hampered by the lack of a comprehensive phytoestrogen database for quantifying exposure. The purpose of this research was to develop such a database for use with food-frequency questionnaires in large epidemiologic studies. METHODS: The database is based on consumption patterns derived from semistructured interviews with 118 African-American, Latina, and white women residing in California's San Francisco Bay Area. HPLC-mass spectrometry was used to determine the content of seven specific phytoestrogenic compounds (i.e. the isoflavones: genistein, daidzein, biochanin A, and formononetin; the coumestan: coumestrol; and the plant lignans: matairesinol and secoisolariciresinol) in each of 112 food items/groups. RESULTS: Traditional soy-based foods were found to contain high levels of genistein and daidzein, as expected, as well as substantial amounts of coumestrol. A wide variety of "hidden" sources of soy (that is, soy protein isolate, soy concentrate, or soy flour added to foods) was observed. Several other foods (such as various types of sprouts and dried fruits, garbanzo beans, asparagus, garlic, and licorice) were also found to be substantial contributors of one or more of the phytoestrogens analyzed. CONCLUSIONS: Databases, such as the one described here, are important in assessing the relationship between phytoestrogen exposure and cancer risk in epidemiologic studies. Agencies, such as the United States Department of Agriculture (USDA), that routinely provide data on food composition, on which epidemiologic investigations into dietary health effects are based, should consider instituting programs for the analysis of phytochemicals, including the phytoestrogens. PMID- 10843441 TI - Sources of phytoestrogen exposure among non-Asian women in California, USA. AB - OBJECTIVE: We recently described the development of a comprehensive database for assessing phytoestrogen exposure in epidemiologic studies. This paper describes the first application of this database and the primary sources of phytoestrogen consumption in non-Asian women. METHODS: Four hundred and forty-seven randomly selected African-American, Latina, and white women, ages 50-79 years, residing in California's San Francisco Bay Area and participating as controls in an ongoing population-based case-control study of breast cancer, were included in the present analysis. Average daily consumption of each of seven phytoestrogenic compounds was determined for each woman by combining the values from the new database with food consumption reported on a food-frequency questionnaire. RESULTS: Phytoestrogens in the non-Asian Bay Area diet appear to come primarily from: (1) traditional soy-based foods (e.g. tofu and soy milk); (2) "hidden" sources of soy (e.g. foods containing added soy protein isolate, soy concentrate, or soy flour, e.g. many brands of doughnuts and white bread); and (3) a variety of foods which contain only low to moderate amounts of phytoestrogens per 100 grams but which are frequently consumed (e.g. coffee and orange juice). CONCLUSIONS: In the absence of a comprehensive assessment of various phytoestrogens in a wide variety of foods, epidemiologic studies could suffer from the effects of uncontrolled confounding by unmeasured sources of phytoestrogen exposure potentially leading to biased estimates of effect and misinterpretation of findings. PMID- 10843442 TI - Allergic disorders and the risk of childhood acute lymphoblastic leukemia (United States). AB - OBJECTIVES: To test the hypothesis that childhood acute lymphoblastic leukemia (ALL) is associated with allergic disorders. METHODS: We compared the histories of selected allergic disorders (asthma, hay fever, food or drug allergies, eczema, and hives) of 1842 cases of ALL with those of 1986 individually matched controls. The histories of the allergic disorders among siblings of cases and controls were also compared. RESULTS: The combined history of any one or more of the five allergic disorders evaluated was associated with a significant reduced risk of ALL (adjusted OR = 0.7, 95% CI 0.6-0.8), as were histories of four specific allergic disorders (asthma, hay fever, food or drug allergies, and eczema). The combined history of any one or more of the five allergic disorders among any of the siblings of the study subjects also revealed a significantly inverse association (adjusted OR = 0.9, 95% CI 0.8-1.0). CONCLUSION: The results from this study, in agreement with most previous studies on adult cancer, suggest that allergic disorders may be associated with a reduced risk of childhood ALL. PMID- 10843443 TI - Future trends in breast, cervical, lung, mouth and pharyngeal cancer incidence in Slovenia. AB - OBJECTIVES: Breast, cervical, lung, mouth and pharyngeal cancers are important public health problems in Slovenia, and in many other Central and South European countries. The aim of this study was to predict the incidence of these cancers in Slovenia up to the year 2009, based on the data of the Cancer Registry of Slovenia for the period 1965-1994 and on the official national population projections for the Republic of Slovenia. METHODS: Age-period-cohort models were applied. In the case of data heterogeneity in lung as well as in mouth and oropharyngeal cancer in males, an additional parameter indicating differences in lifestyle was introduced in the model. RESULTS: After accounting for major site specific risk factors, we predict in females a steady increase in breast and lung cancer, but no major changes in cervical cancer case-load. In males a steady decrease in the lung cancer case-load is expected throughout the predicted period, while for mouth and pharyngeal cancer a moderate decrease is expected only after the year 2000. CONCLUSION: Despite some uncertainties inherent in cancer incidence predictions, the obtained results are important in setting priorities for national cancer control strategies in Slovenia, especially in further efforts towards primary prevention of lung, mouth and pharyngeal cancer, and in more efficient early detection of breast, cervical, mouth and pharyngeal cancer. PMID- 10843444 TI - Cancer risk associated with subfertility and ovulation induction: a review. AB - OBJECTIVE: Over the past decades the use of fertility drugs (FDs) has greatly increased. Recently, the possible association between the use of FDs and risk of cancer has aroused great concern. In this paper, we critically review the available epidemiologic studies. METHODS: We identified papers published between 1966 and 1999 that examined FDs and specific causes of subfertility in relation to the risks of cancers of the ovary, breast, endometrium and thyroid, and melanoma. RESULTS: Although present insights into the pathogenesis of hormone related malignancies suggest a possible association between the use of FDs and the risk of specific cancers, this has not been convincingly demonstrated in epidemiologic studies. With regard to cancer risk in relation to the cause of subfertility, the only consistent association observed is an increased risk of endometrial cancer for women with subfertility due to hormonal disorders. While positive findings in some studies on FDs and ovarian cancer risk have aroused serious concern, the associations observed in most of these reports appear to be due to bias or chance rather than being causal. The most important sources of bias are inadequate confounder control for both parity and causes of subfertility. CONCLUSIONS: To discriminate between the possible carcinogenic effects of various ovulation induction regimens, subfertility disorders, and reproductive characteristics associated with subfertility, future studies should include large populations of subfertile women with sufficient follow-up time. In such cohort studies the cause of subfertility should be measured adequately (based on medical records) and information about reproductive characteristics should be collected for all cohort members. Such studies should also include a group of subfertile women with an indication for FD use but not so treated. PMID- 10843445 TI - Alcohol consumption and breast cancer among black and white women in North Carolina (United States). AB - OBJECTIVE: The purpose of this study was to investigate the effects of alcohol consumption on breast cancer risk in black and white women. METHODS: We used data from the Carolina Breast Cancer Study, a population-based, case-control study of black and white women in North Carolina. Interviews were conducted with 890 cases and 841 controls frequency-matched on age and race. RESULTS: Overall, the prevalence of moderate to high levels of alcohol consumption was low. Compared with abstainers, the multivariate odds ratio for recent intake of one or two drinks per day was 1.4 (95% CI = 0.9-2.1) and two or more drinks a day was 1.0 (95% CI = 0.6-1.6); increasing consumption was not associated with risk (p for trend = 0.6). The associations were similar, but somewhat weaker, for average lifetime consumption. Among women who consumed 91 g/week or more of alcohol, a nonsignificant increased risk of breast cancer was observed for women reporting binge drinking (OR = 1.5; 95% CI = 0.9-2.3), but not for those who consumed less than 91 g/week reporting binge drinking (OR = 1.0; 95% CI = 0.6-1.5). Odds ratios did not differ meaningfully by race, age, menopausal status, exogenous hormone use, or body mass index. CONCLUSIONS: These data provide little evidence for an association between alcohol consumption and risk of breast cancer among either black or white women. PMID- 10843446 TI - Influence of vegetables on the expression of GSTP1 in humans--a pilot intervention study (Sweden). AB - BACKGROUND: There are indications that a diet rich in vegetables and/or fruit has a protective effect against several types of diseases, including cancer. Data from experimental and epidemiological studies suggest that antioxidant constituents may provide protection against environmental carcinogens. AIMS: This study investigated the effect of additional vegetables in the diet on the expression of the endogenous antioxidant enzyme GSTP1 in human lymphocytes. METHODS: Six subjects were given an addition of mixed vegetables to their normal diet for a period of three weeks. The expression of GSTP1 protein and mRNA in lymphocytes was measured by Western blot and RT competitive PCR. RESULTS: After the intervention all six subjects had lower levels of GSTP1 mRNA, and five of the six subjects had lower GSTP1 protein levels. This suggests that increased vegetable intake decreases GSTP1 expression, possibly through the supply of additional antioxidants. PMID- 10843447 TI - Alcohol consumption, smoking and risk of gastric cancer: case-control study from Moscow, Russia. AB - OBJECTIVES: To examine the risk of gastric cancer associated with alcohol consumption and smoking in men and women in Moscow, Russia. MATERIALS AND METHODS: A case-control study which includes 448 cases and 610 controls was conducted. Cases consisted of patients with newly diagnosed histologically confirmed gastric cancer. Controls were patients admitted during the study period to the hospital with diagnoses other than cancer and/or gastrointestinal diseases. Information on demographic variables, smoking, alcohol consumption and diet was collected from all subjects. Venous blood was drawn from 361 cases and 441 controls. A serological test for Helicobacter pylori immunoglobulin G was performed. RESULTS: Alcohol consumption, particularly vodka consumption, was found to increase the risk of gastric cancer. In men the effect of hard liquor drinking was stronger for cancer of the cardia (OR = 3.4, CI = 1.2-10.2), while in women the effect was stronger for cancer of sites other than gastric cardia (OR = 1.5, CI = 1.0-2.3). Smoking increased the risk of developing gastric cancer in men, but not in women. In men a dose-response relationship between mean number of cigarettes smoked per day (p = 0.03), pack-years of cigarettes smoked (p = 0.01) and duration of smoking (p = 0.08) and the risk of cancer of gastric cardia was observed. Further statistical analysis revealed interactions between effect of smoking and alcohol consumption and between smoking and H. pylori infection status. CONCLUSIONS: The findings further support the role of alcohol consumption and smoking in the etiology of gastric cancer. PMID- 10843448 TI - Increased lymphocyte replicative index following 2,4-dichlorophenoxyacetic acid herbicide exposure. AB - OBJECTIVE: Evaluate peripheral blood lymphocyte proliferation (replicative index:RI) and micronuclei frequency (MF) among 2,4-D herbicide applicators. METHODS: Twelve applicators spraying only 2,4-D provided a blood and urine specimen upon enrollment, several urine samples during the spraying season, and a blood specimen at the study's end. Nine controls provided blood and urine specimens upon enrollment and at the study's end. Gas chromatography/tandem mass spectroscopy determined urinary 2,4-D levels and standard in-vitro assays determined RI and MF scores. Applicator RI and MF were compared before and after spraying and with controls. RESULTS: Applicators contributed 45 urine specimens with concentrations ranging from 1.0 to 1700 (microg 2,4-D/g creatinine/L urine) that logarithmically (In) increased as spraying time increased. Applicator RI increased after spraying (p = 0.016), independent of tobacco and alcohol use, and demonstrated a weak dose-response with increasing urinary 2,4-D levels (p = 0.15). Among 2,4-D applicators, pre-exposure complete blood counts and lymphocyte immunophenotypes were not significantly different from post-exposure measurements. CONCLUSION: Urinary 2,4-D concentration, an exposure biomarker, may be associated with lymphocyte replicative index, a cell proliferation biomarker. PMID- 10843449 TI - Divestment in the tobacco industry. PMID- 10843450 TI - Comparison of prazosin, terazosin and tamsulosin in the treatment of symptomatic benign prostatic hyperplasia: a short-term open, randomized multicenter study. BPH Medical Therapy Study Group. Benign prostatic hyperplasia. AB - BACKGROUND: The objective of this open randomized clinical study was to compare the short-term efficacy and safety of three alpha-1 blockers, prazosin, terazosin and tamsulosin, in the treatment of lower urinary tract symptoms (LUTS) associated with benign prostatic hyperplasia (BPH). METHODS: The study comprised 121 patients with symptomatic BPH who were randomized to receive 0.5 mg of prazosin twice daily, 0.5 mg of terazosin twice daily or 0.1 mg of tamsulosin once daily for the initial 2 weeks. The doses were doubled for the next 2 weeks. The primary variables assessed were a symptom score, changes in maximum and average urinary flow rate (Qmax and Qave), postvoid residual urine volume and blood pressure. RESULTS: The percentage changes in the total symptom score from baseline were 38, 39 and 26% at 4 weeks by prazosin, terazosin and tamsulosin, respectively. Terazosin produced significantly higher improvement in four out of nine individual symptoms than tamsulosin (P < 0.05). A significant increase in Qmax or Qave in uroflowmetry was obtained in the prazosin and tamsulosin groups. Blood pressure remained unchanged in normotensive patients, but significantly decreased in hypertensive patients except for the tamsulosin group. Adverse events were minimal in all treatment groups. CONCLUSIONS: The efficacy and safety profiles were different among the alpha-1 blockers at standard doses. Tamsulosin appears to be safer than the others for aged patients or patients with hypertension who have impaired blood pressure regulation, while terazosin is significantly effective in improving symptomatic score when compared with the others examined. It is recommended that the alpha-1 blocking agent and its optimal dose are selected on the basis of the baseline characteristics of the patients with symptomatic BPH. PMID- 10843451 TI - Clinical experience with BCG alone versus BCG plus epirubicin. AB - BACKGROUND: Bacillus Calmette-Guerin (BCG) and epirubicin have both been shown to be effective in the treatment of superficial bladder cancer. We studied whether the alternating combination of these agents could improve the efficacy with tolerable side-effects in the treatment of high-risk superficial bladder tumors. METHODS: Forty-one patients with high-risk superficial bladder transitional carcinoma were included in this study. Twenty-one patients were randomized into the BCG group and 20 patients were treated with sequential BCG and epirubicin. The patients were followed for 9-24 months (mean 18 months). Recurrence rates, median time to the first recurrence, progression rate and complications were compared. RESULTS: Fifteen percent of the patients in the BCG and epirubicin group and 19% of the patients in the BCG alone group developed tumor recurrence. Tumor progression was observed in 4.7% and 10% in the BCG/epirubicin group and the BCG alone group, respectively. Median time to first recurrence was 11 months for the BCG/epirubicin group and 16 months for the BCG group (P > 0.05). Three patients in the BCG/epirubicin treatment group developed serious side-effects, which necessitated antituberculosis treatment. CONCLUSION: Because the efficacy of combination was no better than the standard treatment and the alternating combination seemed to be related to a higher incidence of side-effects, this study albeit small, does not recommend combination therapy of BCG and epirubicin in high risk patients with superficial bladder cancer. PMID- 10843452 TI - Telomerase activity in urine after transurethral resection of superficial bladder cancer and early recurrence. AB - BACKGROUND: To explore the relationship between telomerase activity in urine after transurethral resection (TUR) of superficial bladder cancer and early intravesical recurrence. METHODS: Urine samples were obtained from 42 patients with superficial bladder cancers prior to TUR and on the postoperative day 1 and day 6. These patients were followed-up prospectively by cystoscopy at 3 and 6 months after TUR in combination with urinary cytology and telomerase activity. Telomerase activity in the urine was assessed by the telomeric repeat amplification protocol assay. RESULTS: Urinary telomerase activity prior to TUR was positive in 24 (57%) of the 42 patients. On the postoperative day 1 and day 6, positive urinary telomerase activity was seen in 13 (31%) and nine (21%) patients, respectively. Postoperative urinary telomerase activity on either day 1 or day 6 was significantly associated with pre-operative urinary telomerase activity status (P = 0.0024). Fifteen patients showed intravesical tumor recurrence at 3 months cystoscopic check-up and an additional nine had recurred at the 6 months check-up. Recurrence rate within 6 months in patients with pre operative positive urinary telomerase activity was similar to that in those with negative activity (58.3 vs 58.8%). However, recurrence rate at 3 months for patients with positive activity was higher than that of those with negative activity (50 vs 17.7%), in 23 patients treated only by TUR. CONCLUSIONS: Presence of cells positive for telomerase activity in urine after TUR of superficial bladder cancer indicates persistently existing cancer cells in the urine. It is, however, not a sole predictor of the early intravesical recurrence. PMID- 10843453 TI - Significance of simultaneous determination of serum human chorionic gonadotropin (hCG) and hCG-beta in testicular tumor patients. AB - BACKGROUND: Simultaneous determinations of human chorionic gonadotropin hormone (hCG) and hCG-beta frequently produce discrepancies, that is when hCG or hCG-beta is normal, the other is elevated. Accordingly, we examined the significance of simultaneous determination of serum hCG and hCG-beta in testicular tumors. METHODS: Simultaneous determination of hCG and hCG-beta was performed in 54 patients with testicular seminoma and 74 with non-seminomatous testicular tumors. RESULTS: For detection of seminoma patients, hCG-beta was more effective than hCG because hCG-beta was positive in 83% (45/54) of the patients and hCG was positive in 50% (27/54). In non-seminomatous testicular tumor cases, hCG-beta was positive in 74% (55/74) and hCG was positive in 82% (61/74). The cases of hCG<1.0 mIU/mL and HCG-beta>0.1 ng/mL were significantly more frequently seen in patients with seminoma than in those with non-seminomatous testicular tumor (P < 0.001). Fourteen patients had recurrent tumor. At recurrence, only hCG was elevated in nine cases, only hCG-beta was elevated in two cases and both in one case. For diagnosis of falsely positive hCG, testosterone administration was effective because after testosterone administration, serum hCG levels became undetectable (< 1.0 mIU/mL) within one week in three examined cases. CONCLUSION: Human chorionic gonadotropin-beta was a better marker of seminoma than hCG. For earlier detection of recurrence, both markers should be examined. For diagnosis of falsely positive hCG, testosterone administration was effective. PMID- 10843454 TI - Complicating risk factors for pyelonephritis after extracorporeal shock wave lithotripsy. AB - PURPOSE: The score to predict the risk of post-extracorporeal shock wave lithotripsy (ESWL) pyelonephritis was evaluated. The score was based on the multivariate analysis of risk factors available pre-operatively. Stone size, pyuria, bacteriuria, previous pyelonephritis and other adjunctive procedures had been selected and scored. METHODS: Three-hundred and forty-eight adult patients without active urinary infection undergoing ESWL therapy were studied. One of three regimens were selected by either doctor or patient: (i) no antimicrobial treatment; (ii) one dose of levofloxacin; or (iii) 1 week course of levofloxacin. Who and why selected it were described. Post-ESWL fever over 38 degrees C was defined as the unfavorable event. RESULTS/CONCLUSION: With increasing score, doctors recommend taking an antimicrobial. There were 11 bacteriuric patients and post-ESWL pyelonephritis developed in one of them. Bacteria within the stone and post-ESWL ureteral obstruction caused by the stone fragments were considered to be important in developing pyelonephritis. However, multiple factors were related with it. Although their decision was not based simply on the score, the score was confirmed to be useful in identifying the high-risk patients and, therefore, to implement cost-effective antimicrobial use. PMID- 10843455 TI - Influence of 5-hydroxytryptamine and the effect of a new serotonin receptor antagonist (sarpogrelate) on detrusor smooth muscle of streptozotocin-induced diabetes mellitus in the rat. AB - BACKGROUND: We evaluated the changes in the response of detrusor muscle to 5 hydroxytryptamine (5-HT; serotonin) and its receptor mechanisms in pathologic bladder of diabetes mellitus rats. METHODS: Eight-week-old male Wistar rats were given an intraperitoneal injection of streptozotocin (STZ, 60 mg/kg) in order to induce diabetes mellitus (4 weeks' duration). The bladder strips (10 x 2 mm) were subjected to a tension of 1.0 g in organ baths for the measurements of isotonic contractile responses to 5-HT or relaxation responses to 5-HT antagonists. RESULTS: Bladder strips from STZ-diabetic rats responded to 5-HT with greater contractions than did those of control rats, given vehicle alone. However, bladder strip weights gradually increased in the STZ-diabetic rats as compared with control rats and when the contractile response to 5-HT was expressed as a percentage of the response to 10(-1) mol/L KCl there was no statistically significant difference between the groups. In both control and diabetic rats, the 5-HT3 selective antagonist did not markedly inhibit the contractile response to 5 HT. In contrast, the 5-HT2A selective antagonist inhibited the contractile response to 5-HT in a dose-dependent manner. CONCLUSIONS: These results suggest that the increased contractile response to 5-HT in diabetic rats' bladder is related to smooth muscle hypertrophy and/or hyperplasia and indicate that this effect is mediated by activation of 5-HT2A receptors. PMID- 10843456 TI - Renal cell carcinoma in an L-shaped kidney. AB - A 64-year-old man with a chief complaint of an asymptomatic abdominal mass was diagnosed as having a renal cell carcinoma in his L-shaped kidney. He was successfully treated with partial nephrectomy following selective embolization of the feeder artery. It is thought to be the first reported case of renal cell carcinoma occurring in an L-shaped kidney. PMID- 10843457 TI - Hypercalcemia in a patient with renal cell carcinoma producing parathyroid hormone-related protein and interleukin-6. AB - A patient with renal cell carcinoma who developed humoral hypercalcemia of malignancy is reported. A 52-year-old male patient was diagnosed with renal cell carcinoma and multiple lung metastases. A cell line isolated from the surgical specimen exhibited continuous production of parathyroid hormone-related protein (PTHrP) in vitro. The production of PTHrP from the cancer cells was confirmed by RT-PCR and immunoradiometric assay. The serum calcium level was not enhanced, whereas the lung lesion was developing and producing interleukin-6, a possible modulator of osteoclastic resorption. Hypercalcemia was induced when the PTHrP concentration increased up to 3.3 pmol/L. PMID- 10843458 TI - Choosing the right nonsteroidal anti-inflammatory drug for the right patient: a pharmacokinetic approach. AB - Effective use of the growing number of nonsteroidal anti-inflammatory drugs (NSAIDs), a group that has recently been augmented by the introduction of the selective cyclo-oxygenase-2 inhibitors, requires adequate knowledge of their pharmacokinetics. After oral administration, the absorption of NSAIDs is generally rapid and complete. NSAIDs are highly bound to plasma proteins, specifically to albumin (>90%). The volume of distribution of NSAIDs is low, ranging from 0.1 to 0.3 L/kg, suggesting minimal tissue binding. NSAID binding in plasma can be saturated when the concentration of the NSAID exceeds that of albumin. Most NSAIDs are metabolised by the liver, with subsequent excretion into urine or bile. Enterohepatic recirculation occurs when a significant amount of an NSAID or its conjugated metabolites are excreted into the bile and then reabsorbed in the distal intestine. NSAID elimination is not dependent on hepatic blood flow. Hepatic NSAID elimination is dependent on the free fraction of NSAID within the plasma and the intrinsic enzyme activities of the liver. Renal elimination is not an important elimination pathway for NSAIDs, except for azapropazone. The plasma half-life of NSAIDs ranges from 0.25 to >70 hours, indicating wide differences in clearance rates. Hepatic or renal disease can alter NSAID protein binding and metabolism. Some NSAIDs with elimination predominantly via acylglucuronidation can have significantly altered disposition. Pharmacokinetics are also influenced by chronobiology, and many NSAIDs exhibit stereoselectivity. There appear to be relationships between NSAID concentration and effects. At therapeutically equivalent doses, NSAIDs appear to be equally efficacious. The major differences between NSAIDs are their therapeutic half lives and safety profiles. NSAIDs undergo drug interactions through protein binding displacement and competition for active renal tubular secretion with other organic acids. When choosing the right NSAID for the right patient, individual patient-specific and NSAID-specific pharmacokinetic principles should be considered. PMID- 10843460 TI - Dosage adjustments for antibacterials in obese patients: applying clinical pharmacokinetics. AB - Obesity is associated with physiological changes that can alter the pharmacokinetic parameters of many drugs. Vancomycin and the aminoglycosides are the only antibacterials that have been extensively investigated in the obese population. The apparent volume of distribution (Vd) and total body clearance of vancomycin are increased in obese patients and have a better correlation with total bodyweight (TBW) than with ideal bodyweight (IBW). The Vd of aminoglycosides is increased in obesity and can be estimated from an adjusted bodyweight that accounts for a fraction of the excess bodyweight (TBW - IBW). These observed changes in pharmacokinetic parameters of vancomycin and aminoglycosides in obese patients may necessitate a deviation from the commonly recommended dosages administered to non-obese individuals. There are limited data regarding the pharmacokinetics of other antibacterial classes in obese patients. The available information for cephalosporins suggests that dosages may need to be increased in obese patients in order to obtain similar serum and tissue concentrations as in non-obese patients. Additional pharmacokinetic studies of other antibacterial classes are required in this special patient population. PMID- 10843459 TI - Biotransformation of post-clozapine antipsychotics: pharmacological implications. AB - The need to develop new antipsychotics that have fewer motor adverse effects and offer better treatment of negative symptoms has led to a new generation of drugs. Most of these drugs undergo extensive first-pass metabolism and are cleared almost exclusively by metabolism, except for amisulpride whose clearance is largely due to urinary excretion. Risperidone has metabolic routes in common with ziprasidone but shows differences in regard to other main pathways: the benzisoxazole moiety of risperidone is oxidised by cytochrome P450 (CYP) 2D6 to the active 9-hydroxyrisperidone, whereas the benzisothiazole of ziprasidone is primarily oxidised by CYP3A4, yielding sulfoxide and sulfone derivatives with low affinity for target receptors in vitro. Olanzapine, quetiapine and zotepine also have some common metabolic features. However, for the thienobenzodiazepine olanzapine a main metabolic route is direct conjugation at the benzodiazepine nucleus, whereas for the dibenzothiazepine quetiapine and the dibenzothiepine zotepine it is CYP3A4-mediated oxidation, leading to sulfoxidation, hydroxylation and dealkylation for quetiapine, but N-demethylation to the active nor-derivative for zotepine. Although the promising benzisoxazole (iloperidone) and benzisothiazole (perospirone) antipsychotics share some metabolic routes with the structurally related available drugs, they too have pharmacologically relevant compound-specific pathways. For some of the new antipsychotics we know the isoenzymes involved in their main metabolic pathways and the endogenous and exogenous factors that, by affecting enzyme activity, can potentially modify steady-state concentrations of the parent drug or its metabolite(s), but we know very little about others (e.g. amisulpride isomers, nemonapride). For yet others, information is scarce about the activity of the main metabolites and whether and how these contribute to the effect of the parent drug. Aging reduces the clearance of most antipsychotics, except amisulpride (which requires further evaluation) and ziprasidone. Liver impairment has little or no effect on the pharmacokinetics of olanzapine, quetiapine, risperidone (and 9-hydroxy risperidone) and ziprasidone, but information is lacking for amisulpride. Renal impairment significantly reduces the clearance and prolongs the elimination half life of amisulpride and risperidone. Again, studies are still not available for some drugs (zotepine) and have focused on the parent drug for others (olanzapine, quetiapine, ziprasidone) despite the fact that renal impairment would be expected to lower the clearance of more polar metabolites. Addressing these issues may assist clinicians in the design of safe and effective regimens for this group of drugs, and in selecting the best agent for each specific population. PMID- 10843462 TI - Impact of population pharmacokinetic-pharmacodynamic analyses on the drug development process: experience at Parke-Davis. AB - BACKGROUND: Continued scepticism about the benefits of population pharmacokinetics and/or population pharmacodynamics, here referred to collectively as the population approach, hampers its widespread application in drug development. At the same time the sources of this scepticism have not been clearly defined. In an attempt to capture and clearly define these concerns and to help communicate the value of the population approach in drug development at Parke-Davis we conducted a survey of customers within the company. The results of this survey are presented here. METHODS: All drug development programmes conducted over the past 10 years that included a population approach in data analysis and interpretation were identified. A brief description of the population analysis was prepared together with a brief description of how the resulting information was used in each drug development programme. These synopses were forwarded to relevant members of each drug development team together with a survey designed to solicit opinions as to the relevance and impact of these analyses. RESULTS: The most frequent use of information derived from population based analysis was in labelling. In all cases of drugs making to New Drug Application (NDA) submission the analyses resulted in information that was included in approved or proposed labelling. In almost half of the cases summarised here (5 of 12), population-based analysis was perceived to have resulted in information that influenced the direction of individual development programmes. In many of these cases the information was serendipitous. It is also noted that most of these analyses were not the result of clearly defined objectives and prospective analysis plans. CONCLUSIONS: Use of the population approach, even when applied retrospectively, may have value in complementing or supporting interpretation of other data collected during the course of a trial. Atypical systemic exposure is quickly and easily assessed for correlation with adverse events or exceptional efficacy in retrospective or ad hoc evaluation. Although we know of no direct evidence, it is possible that such use of population pharmacokinetic data has facilitated NDA review and approval by providing insight into the role of atypical systemic drug exposure in otherwise spurious events. PMID- 10843463 TI - The problems of surgical training in gynaecologic oncology. AB - During the late 60s and early 70s it was recognised that the surgical training for residents in Obstetrics and Gynaecology was inadequate to comprehensively acquire the surgical skills necessary in managing women with gynaecologic cancers. Gynaecologic Oncology (Gynae-Oncology) has three important goals: 1) to maintain the highest standards for patients with gynaecologic cancer, 2) to provide the trainee with clinical skills and structural clinical research after his/her surgical completion, and 3) to acknowledge clinical training centers for the trainees in Gynae-Oncology. For women trainees careful family planning, good communication, flexibility from the program director and faculty, support from co workers, and, most important, participation and support from the spouse of the trainee are the ingredients of successful childbearing and family care during training programs. These problems have to be faced and dealt with before they become insurmountable problems. International standards are needed for training programs of trainees in Gynae-Oncology, with special emphasis on surgical skills. PMID- 10843464 TI - The great imitator: miliary peritoneal tuberculosis mimicking stage III ovarian carcinoma. AB - PURPOSE: To report a case of peritoneal tuberculosis initially mistaken at the time of surgery for metastatic ovarian carcinoma. CASE REPORT: A 31-year-old Filipino female was found to have increasing abdominal pain, ascites, early satiety, increasing abdominal girth and bilateral complex masses. Diagnostic laparoscopy revealed the presence of > 4 liters of ascites, a frozen pelvis secondary to what appeared to be bilateral ovarian carcinomas, along with miliary seeding of the entire anterior abdominal wall, omentum, small bowel and small bowel mesentery, right and left diaphragmatic surfaces, and hepatic surface. Frozen section, at the time of laparotomy, revealed necrotizing granulomas most consistent with disseminated tuberculosis that was confirmed at the time of final pathologic review and culture. CONCLUSION: Peritoneal tuberculosis can be mistaken for widely metastatic ovarian carcinoma at time of surgery. PMID- 10843461 TI - Methods for clinical monitoring of cyclosporin in transplant patients. AB - Cyclosporin was introduced into clinical practice in the early 1980s and has since been shown to prolong survival for transplant recipients. Because cyclosporin is a narrow therapeutic index drug and there are significant consequences associated with 'subtherapeutic' and 'supratherapeutic' concentrations, cyclosporin therapy is monitored as part of routine patient follow-up. However, the optimal method for the therapeutic drug monitoring of cyclosporin has yet to be defined. Currently, the most common method involves monitoring pre-dose trough concentrations, but this method is less than ideal. Other methods of monitoring cyclosporin therapy include monitoring the area under the concentration-time curve, limited sampling strategies, monitoring of single concentrations other than troughs and pharmacodynamic monitoring. Bayesian forecasting has been used successfully in clinical practice with other drugs with narrow therapeutic indices. However, few studies are available regarding Bayesian forecasting and cyclosporin. Existing studies are preliminary in nature and involve the old Sandimmun formulation rather than the Neoral formulation. Although these methods show promise, they have not gained widespread acceptance. This is because of their impracticality and the lack of prospective studies comparing other monitoring methods with trough concentration monitoring. Further comparative studies evaluating the impact of the specific monitoring method on definite patient outcomes are warranted. PMID- 10843465 TI - Megestrol-induced clinical adrenal insufficiency. AB - OBJECTIVE: To describe a case of megestrol-induced adrenal insufficiency. METHODS: A patient who received megestrol and then developed pituitary-adrenal axis abnormalities is described. The literature is reviewed. RESULTS: The patient developed megestrol-induced Cushingoid features. She developed adrenal insufficiency when megestrol was stopped. CONCLUSION: Megestrol can induce alterations of the pituitary-adrenal axis in some patients. It is important to consider a diagnosis of adrenal insufficiency in patients with symptoms of fatigue, hypotension, and asthenia who have been treated with megestrol. PMID- 10843466 TI - Successful conservative treatment of endometrial carcinoma permitting subsequent pregnancy: report of two cases. AB - Two women with endometrial carcinoma who wished to preserve their childbearing ability received conservative treatment by medroxyprogesterone acetate (MPA, 600 mg/day for 22 weeks and 29 weeks, respectively). Following regression of endometrial lesions, their infertility was treated by inducing ovulation. Intact pregnancy was diagnosed 13 months and 11 months after completion of the MPA treatment, respectively. One patient had a twin pregnancy and delivered two infants at 35 weeks of gestational age. The other patient delivered a full-term baby. They had no evidence of recurrence 60 months and 31 months after the conservative treatment, respectively. We believe this conservative treatment with progestin may be safely performed for young patients with endometrial cancer who wish to preserve their fertility. PMID- 10843467 TI - Isolated para-aortic node involvement in stage IB/II cervical carcinoma. AB - The aim of this report was to describe exceptional cases of patients treated for stage Ib and II cervical carcinoma with isolated para-aortic node involvement and to deduce therapeutic implications. Between 1985 and 1998, 491 women with stage IB or II cervical carcinoma underwent radical hysterectomy with systematic pelvic and para-aortic lymphadenectomy. Five patients had para-aortic metastatic nodes but no external iliac, obturator or common iliac node involvement. These five patients had a tumor size >3 cm. According to these cases, in patients with bulky cervical carcinoma systematic complete lymphadenectomy should be performed in order to avoid misdiagnosis of para-aortic node involvement. PMID- 10843468 TI - Intra-abdominal desmoplastic small round cell tumor in a 68-year-old female. AB - BACKGROUND: Desmoplastic small round cell tumor (DSRCT) of the peritoneum typically occurs in young adults. The mean age of females with DSRCT is 20 years. We describe a DSRCT with an unusual age of presentation mimicking a metastatic ovarian neoplasm. CASE: A 68-year-old para 4 female presented with abdominal enlargement. Laparatomy showed multiple tumor nodules attached to the peritoneal surface. The tumor was debulked. The histological findings were characteristic for DSRCT. Adjuvantly the patient received cytotoxic chemotherapy but died of recurrent disease 3 months after initial diagnosis. CONCLUSION: DSRCT should be added to the differential diagnosis of unusual gynecologic malignancies in elderly as well as younger females. Identification of DSRCT is important because it can be confused with primary ovarian neoplasms. PMID- 10843469 TI - The value of transvaginal sonography in the preoperative assessment of myometrial invasion in high and low grade endometrial cancer and in comparison to frozen section in grade 1 disease. AB - OBJECTIVE: To evaluate the diagnostic accuracy of preoperative transvaginal sonography (TVS) in the detection of deep myometrial invasion in endometrial cancer cases classified by the grade of disease, and in comparison to frozen section analysis in grade 1 cases. METHODS: In a prospective study, 91 patients with confirmed endometrial carcinoma underwent preoperative TVS for evaluation of myoinvasion. Sonographic results were categorized as superficial (less than or equal to 1/2 myometrial depth) and deep invasion (greater than 1/2 myometrial depth). TAH-BSO followed by retroperitoneal lymph node sampling were performed in all patients with grade 2-3 tumors. In patients with grade 1 disease, the surgical specimen was intraoperatively evaluated by frozen section, and lymph node sampling was carried out if deep invasion was determined. The preoperative sonographic findings and the frozen section results were compared to the final histopathology report of myoinvasion. RESULTS: In 77 of the 91 (84.6%) patients, the sonographic assessment of the depth of myoinvasion was in accord with the final histopathologic findings. TVS demonstrated a sensitivity of 87.8% and a specificity of 82.7% in detecting deep invasion in the entire study group (grade 1-3), with positive and negative predictive values (PPV, NPV) of 74.3% and 92.3%, respectively. TVS in grade 1 cases (n=47) showed a sensitivity of 77.7%, a specificity of 79%, PPV of 46.6% and NPV of 93.7%. TVS in cases with grade 2-3 tumors (n=44) showed a sensitivity of 90%, specificity of 91.6%, PPV of 90% and NPV of 91.6%. Thus, the accuracy of TVS in grade 2-3 cases was superior to that achieved in grade 1 cases (91% vs 78.7%; p=.002). The myometrial invasion was assessed by frozen section in 41 out of 47 patients with grade 1 disease and demonstrated a sensitivity of 85.7%, a specificity of 100%, PPV of 100% and NPV of 97.1%. The specificity (100%) and accuracy (97.5%) of the frozen section were found to be superior compared to that of the TVS (79% and 78.7%) in detecting deep invasion in grade 1 cases (p=.008, p=.005, respectively). No statistically significant difference was found between the sensitivity of either technique. CONCLUSIONS: TVS appeared to be a more accurate method for preoperative assessment of myoinvasion in grade 2-3 endometrial cancer patients compared to grade 1 patients. In grade 1 cases, this method achieved lower accuracy in detecting deep invasion compared to the frozen section analysis. Based on these data, the value of preoperative TVS results as the sole criterion in the decision to perform extensive surgical procedures in grade 1 endometrial cancer is questionable and warrants further evaluation. PMID- 10843470 TI - Descriptive epidemiology of endometrial hyperplasia in patients with abnormal uterine bleeding. AB - PURPOSE OF INVESTIGATION: To evaluate the prevalence and epidemiologic characteristics of endometrial hyperplasias in women with abnormal uterine bleeding. METHODS: We performed a retrospective analysis on data gained from 294 patients with histologically documented endometrial hyperplasia (with or without atypia), detected among 1,469 women who underwent fractional dilatation and curettage in our department due to abnormal uterine bleeding from 1986 to 1998. Epidemiologic characteristics were abstracted from the patients' medical charts. RESULTS: 294/1469 women were found with endometrial hyperplasia (258 without atypia and 36 atypical hyperplasias). Thirty-six of them were under 40 years of age. Four of the detected endometrial hyperplasias progressed to endometrial carcinoma (one with simple hyperplasia, two with complex and one with atypical hyperplasia). Obesity and hypertension were justified as risk factors in our study population. CONCLUSIONS: The prevalence of endometrial hyperplasia according to our data was 20%. There were statistically significant differences in most epidemiologic parameters between the two types of hyperplasia. The progression of four endometrial hyperplasias to endometrial adenocarcinoma indicates the need for intense follow-up even in cases where patients undergo conservative therapy. PMID- 10843471 TI - Serum sICAM-1 (soluble intercellular adhesion molecule-1) and M-CSF (macrophage colony-stimulating growth factor) throughout monitoring of 34 non-serous ovarian cancers. AB - PURPOSE: Evaluation of serum ICAM-1 (soluble intercellular adhesion molecule-1) and M-CSF (macrophage colony-stimulating growth factor) determinations for monitoring patients with non-serous ovarian cancers. METHODS: ELISA assay of sICAM-1 (cut-off 235 ng/ml) and M-CSF (cut-off 450 pg/ml) in 190 blood samples from 34 patients. RESULTS: In pre-treatment sera (n=17), sICAM-1 was over the cut off in 12/17 (70.6%), M-CSF in 14/17 (82.4%) and CA 125 in 12/16 (75%). sICAM-1 was related only to age at diagnosis (p=0.0008). M-CSF was positively correlated to FIGO stage (p=0.04) CA 125 was elevated in 90.9% of adenocarcinomas (p=0.033 vs other). None of the 3 biological markers were related to other clinico pathological criteria. Among disease-free patients, higher median concentrations of sICAM-1 and M-CSF were recorded under adjuvant treatment than without (p=0.014, and p=0.08, respectively). After relapse, the highest levels of sICAM 1, M-CSF and CA 125 were observed in progressive disease (46/53, 86.8% (p=0.014), 51/53 96.2% (p=0.008) and 46/52 88.5% (p>0.05), respectively). CONCLUSION: In non serous ovarian cancers, sICAM-1 although elevated in most cases, is not useful for monitoring. Serum M-CSF is a valuable marker when ovarian tumours do not express CA 125. PMID- 10843472 TI - Apoptosis and apoptosis-related proteins (Fas, Fas ligand, bcl-2, p53) in macrophages of human ovarian epithelial tumors. AB - Apoptosis and the apoptosis-related proteins (ARP) (Fas, Fas ligand (FasL), bcl-2 and p53) were analyzed in macrophages of different human ovarian epithelial tumors. Few macrophages were found in ovaries of women without oncologic disorders. In ovarian benign cysts, macrophagic density reached 4.9+/-1.2 per 50,000 microm2, most were present in lymphoid-macrophagic infiltrates of the sub epithelial stroma (3.7+/-0.5% of the area of a slide), and 23.4% were Fas and FasL positive. In borderline tumors, the expanse of lymphoid infiltrates increased to 15.6% of the area of a slide, and the number of macrophages increased 2.4-fold compared to benign cysts. Of the macrophages, 83-88% expressed Fas and FasL, few had bcl-2 and CD25 receptors, and isolated ones were apoptotic. In carcinomas with high lymphoid-macrophagic infiltration, the infiltrate occupied 17.5% of the slide and macrophages amounted to 12.1+/-1.5/50,000 microm2. Many macrophages were in regions of grouping apoptosis of tumor epithelial cells and significantly fewer expressed Fas, FasL and bcl-2. Macrophages destroyed by apoptosis accounted for 4.6%. In carcinomas with low lymphoid-macrophageal infiltration, the area of the last was 5.1% of the slide. There were 8.6+/-0.8 macrophages/50,000 microm2, mainly at the margins of zones of necrosis and of tumor cells' grouping apoptosis. Extensive macrophagic infiltration into tumor parenchyma is one way by which the host immune system destroys tumors. Fas and FasL appear in macrophages of benign cysts, but in borderline tumors and in carcinomas with low infiltration their concentration increases sharply, to 79.8% and 96.6%, respectively. In 4.5% of these cells, apoptosis of macrophages was seen. The findings suggest that macrophages participate in the transfer of ARP to tumor epithelial cells, thereby inducing their apoptosis, but undergoing the simultaneous apoptosis. PMID- 10843473 TI - Adjuvant CMF-chemotherapy and haemostasis. Effect of "classical" and "modified" adjuvant CMF-chemotherapy on blood coagulation fibrinolysis in patients with breast cancer. AB - Effects of "classical" and "modified" adjuvant CMF-chemotherapy on haemostasis were studied in 22 patients with breast cancer receiving cyclophosphamide (100 mg/m2 p.o.; days 1-14 or 600 mg/m2 i.v.; days 1,8), methotrexate (40 mg/m2 i.v.; days 1,8) and 5-fluorouracil (600 mg/m2 i.v.; days 1,8). Blood collection was done prior to chemotherapy on day 1 and 8. A significant decrease of protein C antigen and activity associated with cumulative effects was observed from day 1 to 8. This effect was similar with "classical" and "modified" CMF-chemotherapy but the reduction of protein C was more pronounced with the oral application of cyclophosphamide. In absence of any significant cumulative decrease of other vitamin K-dependent blood coagulation proteins (factor VII, protein S), the simultaneous decrease of protein C activity and antigen indicates a specific influence of CMF-chemotherapy on vitamin K-dependent protein C-synthesis in the liver. PMID- 10843474 TI - DNA polymorphism analysis of a pure non-gestational choriocarcinoma of the ovary: case report. AB - A 45-year-old nulligravida woman died from carcinoma peritonitis with choriocarcinoma arising in the ovary. This tumor was resistant to chemotherapy after debulking surgery. DNA polymorphism analysis was useful in proving the choriocarcinoma to be non-gestational carcinoma. In this paper, the clinical course and DNA polymorphism findings are mainly discussed. PMID- 10843475 TI - Squamous intraepithelial lesions (SILs) and HPV associated changes in HIV infected women or at risk of HIV. DIANAIDS Cooperative Study Group. AB - OBJECTIVES: The study concerns the prevalence of squamous intraepithelial lesions (SILs) and the specific cervical cytopathological features of a group of HIV positive and a group of HIV-negative women recruited in a multicentric cohort study. The assessment of HPV-DNA genotypes was carried out in both groups. METHOD: 459 women, 266 HIV-positive and 193 HIV-negative women at risk were examined in an Italian multi-institutional study involving 14 gynaecological centres. RESULTS: In our samples, the risk of SILs was 29.4% for HIV-positive women and 10% for HIV-negative women (O.R. = 3.90, C.I. 95%: 2.20-6.98) while HPV DNA-PCR genotypes had a high prevalence in both groups of HIV-positive and HIV negative women. Cytopathological features of SILs in HIV-positive women were: a higher number of koilocytes and a more marked atypia of high grade neoplastic cells. CONCLUSIONS: A higher prevalence of SILs as well as a specific cervical cytopathology might suggest HIV infection. PMID- 10843476 TI - Heterologous mullerian mixed tumor after whole body irradiation because of Hodgkin's disease in stage IV. AB - PURPOSE: With an incidence of 1.5% of all malignant diseases of the uterus as specified in the literature, the mullerian mixed tumor is a rarity amongst the malignancies of the female genital tract. METHODS: A retrospective analysis of an individual case report with the occurrence of heterologous mullerian mixed tumor years after irradiation because of Hodgkin's disease. RESULTS: This case reports describes the occurrence of a mullerian mixed tumor 12 years after the treatment of Hodgkin's disease by whole body irradiation. To our knowledge, the incidence of a mullerian mixed tumor after the treatment of Hodgkin's disease has rarely been described up to now in the literature. CONCLUSION: This case report appears to indicate the possible carcinogenic potency of radiotherapy when administered many years before. A causal connection between the administration of whole body irradiation and the development of a mullerian mixed tumor cannot be established. PMID- 10843478 TI - Colposcopy, cytology and HPV-DNA testing in HIV-positive and HIV-negative women. AB - In this study we examined the incidence of colposcopic-colpocytologic findings and analyzed Human Papilloma Virus (HPV)-DNA testing by Polymerase Chain Reaction (PCR) in 104 Human Immunodeficiency Virus (HIV) serous positive women (Group 1) and 218 HIV-negative women (control Groups 2 and 3). The aim of the study was to evaluate the most appropriate and efficacious diagnostic methods for screening programs for cervical cancer in HIV-positive women. For Group 1 we also considered the value of CD4+ T-lymphocytes and morphologic and molecular follow up from 3 to 6 months. The results showed that the abnormal transformation zone (ANTZ) was present in 66.3% of the cases in Group 1 compared with 31.4% in control-Group 2 (p<0.001), and with 58.93% of the cases in control-Group 3 (p=0.257); intraepithelial squamous lesions (SIL) were found in 50% vs 5.66% (p<0.001) and vs 56.25% of the cases (p=0.433), respectively. In 28.85% of the HIV-positive patients the first cytological screening exam was not evaluable due to inflammation but in 56.67% of the cases colposcopy revealed ANTZ. The subsequent colpocytological checkup after therapy showed 10 cases (30%) of low risk squamous intraepithelial lesions (LSIL) and two cases (6.6%) of high risk squamous intraepithelial lesions (HSIL). HPV-DNA testing by PCR was positive in 53.8% of the cases in Group 1, in 6.6% in control-Group 2 and in 42% in control Group 3. In HIV-positive patients multiple HPV genotypes were simultaneously present in 21.43% of the cases and high risk genotypes were present in 70% of the cases of HSIL. In Group 1, 36.61% of the cases had lesions of the lower genital tract. The value of CD4+ T-lympocytes was <200 cells/ml in 30% of the cases of HSIL. Our data, like those of other Authors, confirm a high incidence of HSIL, abnormal colposcopic findings, and HPV infections in HIV-positive women with respect to control-Group 2, while there was not much difference between Group 1 and control-Group 3. Such frequency again suggests that an integrated morphological diagnostic approach with colposcopy-colpocytology in the screening of immunosuppressed subjects would be worthwhile. PMID- 10843477 TI - Factors influencing period of time between negative second-look laparotomy and ovarian carcinoma recurrence. Multicenter study in Poland. AB - OBJECTIVE: The aim of our study was to evaluate factors such as primary clinical stage, presence of ascites, serum CA 125 antigen level, histological type of ovarian cancer, cell differentiation and number of chemotherapy cycles influencing the time of recurrence after negative second-look operations. MATERIAL AND METHODS: Having observed complete clinical remission in 356 patients with ovarian cancer, second-look laparotomy was performed. In 180 patients complete pathologic remission was detected and in 73 recurrence was observed. Correlation analysis between time of recurrence and the above-mentioned prognostic factors was carried out by means of the Mann-Whitney and Kruskal Wallis tests. RESULTS: The time from the second-look operation till diagnosis of relapse ranged from 7 to 36 months (average 21 months). The statistical analysis showed a correlation between the presence of ascites, increased serum CA 125 antigen level, the administration of six chemotherapy courses and the time of recurrence. In all those cases relapse occurred earlier than in patients without ascites, with normal CA 125 antigen levels and after ten courses of chemotherapy. CONCLUSION: Our findings suggest that the stage of clinical advancement and histologic grading do not influence the time of recurrence. The presence of ascites, increased serum CA 125 antigen level and the administration of fewer chemotherapy courses (6 versus 10) after primary surgery affects the earlier relapse of disease. PMID- 10843479 TI - Invasive adenocarcinoma of mixed endocervical and clear cell type, associated with invasive squamous cell carcinoma of the cervix uteri. A case report. AB - A case of invasive adenocarcinoma (predominantly of endocervical type but also with clear cell elements blended in part of the growth) associated with squamous cell carcinoma of the uterine cervix is reported in a young woman aged 18 years. Malignant squamous and atypical glandular cells were detected in Papanicolaou (Pap) smears, which raised the question of a mixed tumor. A diagnosis of cervical cancer was justified by means of a punch biopsy and the patient was treated by modified radical hysterectomy. Histologic and immunohistochemical examination of the hysterectomy specimen revealed two distinct types of an invasive malignant tumor, i.e., of glandular and squamous cell origin, closely related but not integrated. Interestingly, the glandular component comprised both endocervical and clear cell elements. Careful consideration of squamous or glandular atypia in cytological smears may lead to a more precise diagnosis, especially in young women. PMID- 10843480 TI - Relationship between HPV infection of the cervix and blood serum levels of steroid hormones among pre- and postmenopausal women. AB - Human papillomaviruses (HPVs) are frequently present in anogenital lesions but little is known about their role in carcinogenesis. There are steroid hormone response elements in virus genomes that influence expression patterns of viral genes. Activity of the elements may contribute to development of neoplasia in case of hormone level anomalies. Our study was to determine whether the presence of HPV DNA in cervical smears correlates with abnormal levels, of steroid hormones in blood serum. One hundred women aged 40-62 participated in the tests and were divided into two groups: premenopausal and postmenopausal (45 and 55 individuals, respectively). Presence of HPV DNA in cervical smears was detected by PCR and Southern blot hybridisation. Progesterone and estradiol levels in blood serum were measured by radioimmunoassay. Our study showed a higher prevalence of HPV DNA in women with higher levels of progesterone in blood serum. A relationship between hormone level and HPV DNA prevalence should alert clinicians about using hormone contraceptives and hormone replacement therapy. PMID- 10843481 TI - Endometrial polyps: prevalence, detection, and malignant potential in women with abnormal uterine bleeding. AB - PURPOSE OF INVESTIGATION: To report our evaluation of the prevalence and malignant potentiality of endometrial polyps in women with abnormal uterine bleeding, as well as the efficacy of transvaginal ultrasonography and sonohysterography as diagnostic techniques. METHODS: Fractional dilatation and curettage (D&C) was performed in 1,415 patients aged 23-85 years treated in our clinic for abnormal uterine bleeding from 1986 to 1998. Transvaginal ultrasonography was performed prior to D&C on all patients. Sonohysterography was performed only on 157 patients. Diagnostic efficacy for both techniques was evaluated for the detection of endometrial polyps combined with hyperplasia due to sonographic and histologic difficulties in distinguishing them. RESULTS: The prevalence of endometrial polyps was 8.9% (126/1,415). From all 126 endometrial polyps found, 94 were benign, 30 (23.8%) were found with premalignant changes (complex and atypical hyperplasias) and two (1.5%) had undergone malignant degeneration. Transvaginal ultrasonography was of limited diagnostic value for polyps and hyperplasia in premenopausal women, while in postmenopausal women the method provided a greater yield. Sonohysterography was found to be a more effective diagnostic tool. CONCLUSION: Endometrial polyp prevalence rises by age and/or menopause. Malignant degeneration of endometrial polyps was observed only in postmenopausal women. Sonohysterography represents an improvement over conventional ultrasonography and both methods could be used for screening purposes especially when hysteroscopy can not be performed. PMID- 10843482 TI - Gestational trophoblastic disease: the significance of vaginal metastases. AB - PURPOSE: To evaluate patients with vaginal lesions in gestational trophoblastic disease and determine prognostic and therapeutic implications applicable to management. METHODS: Twelve patients among 75 cases of gestational trophoblastic neoplasia were analyzed retrospectively between 1990 and 1997. Vaginal metastases were documented by physical examination and biopsy. Two patients received MAC III regimen (5 and 7 courses), 4 patients received EMA-CO regimen for 2 to 11 courses, while 6 were administered methotrexate alone. Remission was defined as 3 weekly beta hCG levels below assay sensitivity (<5 mIU/ml). RESULTS: The mean age of the patients was 25.4 years. While 10 of the patients presented initially with hemorrhage and bloody leukorrhea, the remaining 2 were diagnosed during a routine study of hydatidiform mole. The sites of involvement were almost always the anterior distal vaginal wall. Five cases had additional lung and 1 case had lung, liver, spleen and brain metastases. Three of the patients who received methotrexate as monotherapy did not respond to therapy and were switched to EMA CO. Overall survival was 91.6%. One patient died in the first month of the initial therapy. CONCLUSION: The presence of large vaginal metastases should be classified as a high-risk factor and these patients must be treated by multiple agent chemotherapy. PMID- 10843483 TI - Is potential to produce inhibins related to prognosis in ovarian granulosa cell tumors? AB - OBJECTIVE: The prognosis of ovarian granulosa cell tumors is difficult to predict by conventional histological methods. STUDY DESIGN: The tumors of 30 patients operated on for a granulosa cell tumor were retrospectively studied for the expression of the inhibin alpha-subunit. The immunohistochemical staining results were correlated to the FIGO stage of the tumor and the prognosis of the patients. RESULTS: Twenty-six (87%) of the ovarian granulosa cell tumors stained positively for the inhibin alpha-subunit. All FIGO stage I and II tumors expressed inhibin alpha-subunit. All FIGO stage I and II tumors expressed inhibin, whereas the majority of stages III and IV did not (p=0.001, chi2-test). The survival of the patients with inhibin immunopositive tumors was significantly longer (median 183 months, SE 41, p=0.000, log rank test) than that of patients without inhibin expression (median 2.5 months, SE 5.25). CONCLUSIONS: Patients with ovarian granulosa cell tumors with a potential to produce the inhibin alpha-subunit may have a more favorable prognosis than those with inhibin immunonegative tumors. PMID- 10843484 TI - Cutaneous fixed drug eruption to paclitaxel; a case report. AB - The aim of this report was to investigate an adverse effect of paclitaxel. Cutaneous fixed drug eruption against paclitaxel--which is rarely seen--is explained. A 70-year-old caucasian woman experienced three operations for ovarian carcinoma and finally a paclitaxel-based chemotherapy regimen was initiated as adjuvant chemotherapy. Following the administration of the first dose of paclitaxel, the patient had a cutaneous reaction that was clinically consistent with "fixed drug eruption". Lesions regressed with a topical steroid dressing. Histopathological examination of the lesion confirmed clinical diagnosis. It is interesting that such a reaction could occur despite the use of premedication with systemic corticosteroid antihistaminic prophylaxis yet heal rapidly with topical corticosteroid therapy. Thus we recommend careful observation for cutaneous side-effects in patients who receive paclitaxel or other taxanes, even with premedication prophylaxis. PMID- 10843485 TI - Hormone replacement therapy after epithelial ovarian cancer treatment. AB - PURPOSE: Our report deals with the presumed influence of hormone replacement therapy (HRT) on the recurrence of epithelial ovarian cancer. MATERIALS AND METHODS: Our study group consisted of 31 patients who had been treated for invasive epithelial carcinoma of the ovaries. In all the patients the primary treatment was surgery. The mean duration of follow-up was 55 months. All the patients received HRT with non-conjugated estrogens. The data were analyzed using analytical descriptive epidemiological methods. RESULTS: The mean duration of HRT was 25 months, starting on average 18 months after completed therapy for ovarian cancer. Progression of the disease was established in 3 patients with advanced disease, 2 patients with moderately- and poorly-differentiated serous adenocarcinoma respectively, and in one patient with well-differentiated endometrioid adenocarcinoma. The progression of ovarian cancer occurred 1, 2 and 10 months after the beginning of HRT. Two patients died due to progressive disease, while one patient is still alive with evidence of the disease. Eleven months after the beginning of HRT, one patient without evidence of ovarian cancer progression presented with a new primary cancer--carcinoma of the breast. CONCLUSIONS: According to the results of our review study, HRT does not seem to have a noteworthy effect on the progression of epithelial carcinoma of the ovary. However, only further--carefully designed--prospective studies could provide more conclusive results. PMID- 10843486 TI - The utilization of immunohistochemical prognostic factor in endometrial adenocarcinoma: is it cost effective? AB - This study investigated the relation between immunohistochemical prognostic factors and clinical stage and histopathological grade in endometrial adenocarcinoma. Twenty-seven patients with a mean age of 61 (38-74), who underwent radical surgery due to endometrial adenocarcinoma in our hospital between 1983-1998, were re-evaluated. For clinical staging FIGO criteria were used. Histopathological differentiation of the tumor was graded as good (grade 1), moderate (grade 2), and poor (grade 3). Estrogen and progesterone receptors, c-erb B2, UEA 1, Ki-67, PCNA and p53 were studied as immunohistochemical prognostic factors. There were no patients in stages IA and IIIB. Among the prognostic factors, PCNA was the most significantly stained marker, followed by c erb B2, estrogen and progesterone receptors, regardless of the clinical stage and histopathological grade of the tumor. The least positivity was achieved with Ki 67. There was no significant difference when each prognostic factor was analysed with respect to clinical stage and histopathological grade. In our study no significant relation was found between the prognostic factors and the clinical stage and histopathological differentiation of the tumor. Therefore the cost effectiveness of the utilization of these factors should be reconsidered. PMID- 10843487 TI - Immunohistochemical staining properties of PCNA, Ki-67, p53, beta-hCG and HPL in trophoblastic disease. AB - OBJECTIVE: In this study proliferating markers PCNA (proliferating cell nuclear antigen) Ki-67 and mutation of supressor gene p53 were investigated in gestational trophoblastic disease (GTL). These markers were tested by using immunostaining with beta subunits of human chorionic gonadotropin (hCG) and human placental lactogen (HPL). MATERIAL AND METHODS: Twenty curetting samples, 20 spontaneous abortions, 16 hydatidiform moles and two choriocarcinomas were studied and compared. Hydatidiform moles were subdivided into 10 complete and six partial moles by using flow cytometry analysis. All slides were stained with PCNA, Ki-67, p53, hCG, and HPL immunohistochemically. PCNA and Ki-67 stained slides were studied quantitatively to determine the PCNA and Ki-67 index. Other slides that were stained with p53, hCG, HPL were evaluated according to staining percentage and intensity. Staining properties of all groups were compared with each other. Variance analysis and the Mann Whitney U test were used for statistical analysis. Choriocarcinomas were not included in the statistical analysis. Ki-67 and the PCNA index in two choriocarcinoma cases found 81.4% and 41%, and 44% and 64%, respectively. One case was stained in 70% with (++) intensity by p53. While both were stained in 80% with (++) intensity by hCG, one was stained in 30% field (+) intensity by HPL. RESULTS: The four groups of complete and incomplete diagnosed hydatiform moles, spontaneous abortions and retention curettage were matched in pairs and evaluated according to the PCNA index. This index showed significant differences among the groups. The differences among the Ki-67 index, p53, hCG and HPL staining properties were not statistically significant. CONCLUSION: Our findings showed that PCNA is a significant and useful marker for trophoblastic diseases and can be used as a prognostic factor. PMID- 10843488 TI - Clinico-pathological profile of carcinoma of the vulva. AB - There is a need for patient and physician education with regard to the early diagnosis of carcinoma of the vulva. Physicians must be made more aware of the importance of taking a biopsy diagnosis before treating a vulvar lesion. Biopsy of the vulva is a simple procedure that can be performed in any physician's office. Now we must all shift to the 3-incision operative procedure as it has proven to have less morbidity & mortality. PMID- 10843489 TI - Synthesis and properties of uniquely modified oligoribonucleotides: yeast tRNA(Phe) fragments with 6-methyluridine and 5,6-dimethyluridine at site-specific positions. AB - The phosphoramidites of 6-methyluridine and 5,6-dimethyluridine were synthesized and the modified uridines site-selectively incorporated into heptadecamers corresponding in sequence to the yeast tRNA(Phe) anticodon and TpsiC domains. The oligoribonucleotides were characterized by NMR, MALDI-TOF MS and UV-monitored thermal denaturations. The 6-methylated uridines retained the syn conformation at the polymer level and in each sequence location destabilized the RNAs compared to that of the unmodified RNA. The decrease in RNA duplex stability is predictable. However, loss of stability when the modified uridine is in a loop is sequence context dependent, and can not, at this time, be predicted from the location in the loop. PMID- 10843490 TI - Influence of diastereomeric ratios of deoxyribonucleoside phosphoramidites on the synthesis of phosphorothioate oligonucleotides. AB - Extensive investigations on the influence of diastereomeric ratios of deoxyribonucleoside phosphoramidites on stereo-reproducibility of solid phase synthesis of phosphorothioate oligodeoxyribonucleotides via the phosphoramidite approach indicate that the process is stereoreproducible and under inherent process control. PMID- 10843491 TI - Development of a purification procedure for the isolation of nucleosides from urine prior to mass spectrometric analysis. AB - A chromatographic separation of nucleosides from urine has been developed in order to facilitate their mass spectrometric analysis for clinical diagnosis. A number of chromatographic resins were studied in order to develop an effective and efficient purification procedure. The optimized sequential protocol comprises a centrifugation, acidification and neutralization step, followed by application of an affinity chromatographic column and finally further separation on an acidic cation exchange column and a basic anion exchanger. This scheme shows effective clean-up of a standard radiolabelled nucleoside with a recovery of 92.5%, and recovery of nucleosides added to urine samples before extraction showed recoveries of 72-82%. PMID- 10843492 TI - A convenient synthesis of 2'-deoxy-2-fluoroadenosine; a potential prodrug for suicide gene therapy. AB - A convenient synthesis of 2'-deoxy-2-fluoro-adenosine (1) is described. Deaminative fluorination of 2-aminoadenosine (2) followed by silylation of the 3', 5'-hydroxyl groups gave the corresponding 2-fluoroadenosine derivative 4 in good yield. Thiocarbonylation of 4 to thiocarbonylimidazolyl derivative 5a followed by treatment with an excess of tris(trimethylsilyl)silane (TTMSS) and tert-butyl peroxide in toluene at 80 degrees C was found to affect an efficient deoxygenation to the corresponding 2'-deoxy derivative 6. Desilylation of 6 by Et4NF in CH3CN afforded 1 in high yield. PMID- 10843493 TI - Synthesis and biological activities of new carbaacyclonucleosides and 1' oxaacyclonucleosides related to clitocine. AB - We describe the synthesis of two series of acyclonucleosides:carbaacyclonucleosides and 1'-oxaacyclonucleosides which possess the same aglycone as clitocine 3 which is a natural nucleoside exhibiting interesting biological properties. These compounds have been obtained by condensation of 4-aminobutanol or 3-silyloxypropoxyamine with 4,6-dichloro-5 nitropyrimidine. Structural modifications have been made on the heterocyclic base and the side chain to enhance their potential activity. All these compounds have been tested against different viruses: HIV-1, HSV-1, HSV-2, CMV, VZV, EBV. The carbaacyclonucleoside 10 was associated with an anti-EBV activity (EC50 = 0.86 microg/mL). PMID- 10843494 TI - Formation and reactivity of 2,4-ditriazolyl pyrimidine C-nucleoside derived from pseudouridine. AB - Thymine and 2',3',5'-tri-O-acetyl-psi-uridine (1) was converted into the corresponding 2,4-ditriazolyl derivatives 5 and 2, respectively. Of these two substituents, the C4-triazolyl group was found to be quite susceptible to nucleophilic substitution while the other triazolyl is resistant. PMID- 10843495 TI - Synthesis of oligodeoxyribonucleotide bearing 2'-S-alkyl residue and its effect on the duplex stability. AB - 2'-Deoxy-2'-S-hexyluridine derivative was synthesized from 2,2'-anhydrouridine and 1-hexanethiol and incorporated into an oligodeoxyribonucleotide. The thermal stability of the duplexes formed by the 2'-S-hexyl modified ODN with either the complementary DNA or RNA strand was decreased compared to the unmodified counterparts. PMID- 10843496 TI - Synthesis and biological evaluation of 1,3-oxathiolane 5-azapyrimidine, 6 azapyrimidine, and fluorosubstituted 3-deazapyrimidine nucleosides. AB - (2R,5S)-5-Amino-2-[2-(hydroxymethyl)-1,3-oxathiolan-5-yl]- 1,2,4-triazine-3(2H) one (8) and (2R,5R)-5-amino-2-[2-(hydroxymethyl)-1,3-oxathiolan-5-yl]-1,2,4-tr iazine-3(2H)-one (9) have been synthesized via a multi-step procedure from 6 azauridine. (2R,5S)-4-Amino-1-[2-(hydroxymethyl)-1,3-oxathiolan-5-yl]-1,3, 5 triazine-2(1H)-one (11) and (2R,5R)-4-amino-1-[2-(hydroxymethyl)-1,3-oxathiolan-5 yl]- 1,3,5-triazine-2(1H)-one (12), and the fluorosubstituted 3-deazanucleosides (19-24) have been synthesized by the transglycosylation of (2R,5S)-1-[2-[[(tert butyldiphenylsilyl) oxy]methyl]-1,3-oxathiolan-5-yl] cytosine (2) with silylated 5-azacytosine and the corresponding silylated fluorosubstituted 3-deazacytosines, respectively, in the presence of trimethylsilyl trifluoromethanesulfonate as the catalyst in anhydrous dichloroethane, followed by deprotection of the blocking groups. These compounds were tested in vitro for cytotoxicity against L1210, B16F10, and CCRF-CEM tumor cell lines and for antiviral activity against HIV-1 and HBV. PMID- 10843498 TI - Stereoselective synthesis and X-ray structure of 1-(4-O-benzyl-1,5-anhydro-2,3 dideoxy-D-arabino-hex-1-enitol-3-yl )-4-me thoxy-1H-pyrimidin-2-one. AB - The intramolecular glycosylation of a 6-O-(4-methoxypyrimidin-2-yl) derivative of phenyl 2,3-didehydrohex-2-eno-1-thiopyranoside afforded 3-deoxy-3-(4 methoxypyrimidin-2-on-1-yl)glycal as the major product in a stereoselective manner. The isolated 3'-deoxyglycal nucleoside was characterized by X-ray and NMR spectral analyses. PMID- 10843497 TI - Efficient synthesis of carbocyclic nucleoside, (+/-)-homocarbovir via pi allylpalladium complex formation from the allyl-N,N-ditosylimide substrate. AB - The synthesis of a carbovir analogue, (+/-)-homocarbovir (3) was achieved from norbornadiene (4) in seven steps and 27% overall yield. This route involves a Meinwald-type rearrangement, an acid-hydrolysis of N-tosyl bicyclic enamine 5, and a Pd(0)-catalyzed coupling reaction. PMID- 10843499 TI - Syntheses, crystal structures, and hydrogen bonding patterns of 3'-C methylenecarboxylic-3'-deoxythymidine and 3'-C-methyleneamidilylic-3' deoxythymidine. AB - Thymidine derivatives containing carboxylic acid and amide groups have been synthesized and the hydrogen-bonding patterns of 3'-C-methylenecarboxylic-3' deoxythymidine 6 and 3'-C-methyleneamidilylic-3'-deoxythymidine 9 have been characterized by using X-ray crystallography. PMID- 10843500 TI - The latest progress in the synthesis of carbocyclic nucleosides. AB - This review presents the latest developments in the field of carba-nucleosides (1994-1998). Special attention is paid to the synthesis of key precursors to those carba-nucleosides that possess significant biological activities or have novel structures. PMID- 10843501 TI - Cellular pharmacology of the D- and L-enantiomers of beta-5-o-carboranyl-2' deoxyuridine. AB - The cellular pharmacology of the D- and L-enantiomers of beta-5-o-carboranyl-2' deoxyuridine (CDU), compounds designed for boron neutron capture therapy (BNCT), were studied using human CEM lymphoblast and U-251 glioblastoma cells, at a physiologically achievable concentration (1 microM). Accumulation of the enantiomers was rapid and indistinguishable, reaching cellular concentrations > 40-fold higher than extracellular levels, with approximately 5% persisting in cells after incubation in fresh medium for more than 2 hr. Uptake was not affected by nucleoside uptake inhibitors, but was inhibited by the purine base uptake inhibitor papaverine. PMID- 10843502 TI - Surveillance for waterborne-disease outbreaks--United States, 1997-1998. AB - PROBLEM/CONDITION: Since 1971, CDC and the U.S. Environmental Protection Agency (EPA) have maintained a collaborative surveillance system for collecting and periodically reporting data relating to occurrences and causes of waterborne disease outbreaks (WBDOs). REPORTING PERIOD COVERED: This summary includes data from January 1997 through December 1998 and a previously unreported outbreak in 1996. DESCRIPTION OF THE SYSTEM: The surveillance system includes data regarding outbreaks associated with drinking water and recreational water. State, territorial, and local public health departments are primarily responsible for detecting and investigating WBDOs and voluntarily reporting them to CDC on a standard form. RESULTS: During 1997-1998, a total of 13 states reported 17 outbreaks associated with drinking water. These outbreaks caused an estimated 2,038 persons to become ill. No deaths were reported. The microbe or chemical that caused the outbreak was identified for 12 (70.6%) of the 17 outbreaks; 15 (88.2%) were linked to groundwater sources. Thirty-two outbreaks from 18 states were attributed to recreational water exposure and affected an estimated 2,128 persons. Eighteen (56.3%) of the 32 were outbreaks of gastroenteritis, and 4 (12.5%) were single cases of primary amebic meningoencephalitis caused by Naegleria fowleri, all of which were fatal. The etiologic agent was identified for 29 (90.6%) of the 32 outbreaks, with one death associated with an Escherichia coli O157:H7 outbreak. Ten (55.6%) of the 18 gastroenteritis outbreaks were associated with treated pools or ornamental fountains. Of the eight outbreaks of dermatitis, seven (87.5%) were associated with hot tubs, pools, or springs. INTERPRETATION: Drinking water outbreaks associated with surface water decreased from 31.8% during 1995-1996 to 11.8% during 1997-1998. This reduction could be caused by efforts by the drinking water industry (e.g., Partnership for Safe Water), efforts by public health officials to improve drinking water quality, and improved water treatment after the implementation of EPA's Surface Water Treatment Rule. In contrast, the proportion of outbreaks associated with systems supplied by a groundwater source increased from 59.1% (i.e., 13) during 1995-1996 to 88.2% (i.e., 15) during 1997-1998. Outbreaks caused by parasites increased for both drinking and recreational water. All outbreaks of gastroenteritis attributed to parasites in recreational water were caused by Cryptosporidium, 90% occurred in treated water venues (e.g., swimming pools and decorative fountains), and fecal accidents were usually suspected. The data in this surveillance summary probably underestimate the true incidence of WBDOs because not all WBDOs are recognized, investigated, and reported to CDC or EPA. ACTIONS TAKEN: To estimate the national prevalence of waterborne disease associated with drinking water, CDC and EPA are conducting a series of epidemiologic studies to better quantify the level of waterborne disease associated with drinking water in nonoutbreak conditions. The Information Collection Rule implemented by EPA in collaboration with the drinking water industry helped quantifythe level of pathogens in surface water. Efforts by CDC to address recreational water outbreaks have included meetings with the recreational water industry, focus groups to educate parents on prevention of waterborne disease transmission in recreational water settings, and publications with guidelines for parents and pool operators. PMID- 10843503 TI - Prevalence of cigarette smoking among secondary school students--Budapest, Hungary, 1995 and 1999. AB - The average per capita cigarette consumption in Hungary is among the highest in the world (World Health Organization [WHO], unpublished data, 1997) (1). In 1999, the Metropolitan Institute of State Public Health and Public Health Officer Service, Budapest, Hungary, collaborating with CDC, conducted a survey of cigarette smoking among secondary school students aged 14-18 years in Budapest (1999 population of Budapest: approximately 2 million), similar to a survey conducted in 1995 (2). This report summarizes the survey findings, which indicate that current smoking among secondary school students in Budapest increased from 36% in 1995 to 46% in 1999. PMID- 10843504 TI - Costs of smoking among active duty U.S. Air Force personnel--United States, 1997. AB - Smoking is the leading cause of preventable disease and death in the United States (1). The health consequences of smoking impose a substantial economic toll on persons, employers, and society. Smoking accounts for $50-$73 billion in annual medical-care expenditures, or 6%-12% of all U.S. medical costs (2-5). The costs associated with lost productivity also are extensive (2). In 1997, approximately 25% of male and 27% of female active duty Air Force (ADAF) personnel aged 17-64 years were smokers (6). A 1997 retrospective cohort study was conducted among ADAF personnel to estimate the short-term medical and lost productivity costs of current smoking to the U.S. Air Force (USAF). This report summarizes the results of the study, which indicate that current smoking costs the USAF approximately $107.2 million per year: $20 million from medical-care expenditures and $87 million from lost workdays. PMID- 10843505 TI - Progress toward poliomyelitis eradication--African Region, 1999-March 2000. AB - In 1988, the World Health Assembly resolved to eradicate poliomyelitis globally by 2000 (1). The African Region (AFR) of the World Health Organization (WHO) began implementing polio eradication strategies in 1996, including National Immunization Days (NIDs*) and acute flaccid paralysis (AFP) surveillance (2,3). This report summarizes progress toward polio eradication in AFR during 1999-March 2000, and suggests that although substantial progress has been reported toward interrupting poliovirus transmission in eastern and southern Africa, poliovirus remains endemic in other African countries in west and central Africa, especially among those experiencing internal strife or civil war. PMID- 10843506 TI - To measure physical performance in many sports is often very easy and is also done with great precision. PMID- 10843507 TI - Physiological models to understand exercise fatigue and the adaptations that predict or enhance athletic performance. AB - A popular concept in the exercise sciences holds that fatigue develops during exercise of moderate to high intensity, when the capacity of the cardiorespiratory system to provide oxygen to the exercising muscles falls behind their demand inducing "anaerobic" metabolism. But this cardiovascular/anaerobic model is unsatisfactory because (i) a more rigorous analysis indicates that the first organ to be affected by anaerobiosis during maximal exercise would likely be the heart, not the skeletal muscles. This probability was fully appreciated by the pioneering exercise physiologists, A. V Hill, A. Bock and D. B. Dill, but has been systematically ignored by modern exercise physiologists; (ii) no study has yet definitely established the presence of either anaerobiosis, hypoxia or ischaemia in skeletal muscle during maximal exercise; (iii) the model is unable to explain why exercise terminates in a variety of conditions including prolonged exercise, exercise in the heat and at altitude, and in those with chronic diseases of the heart and lungs, without any evidence for skeletal muscle anaerobiosis, hypoxia or ischaemia, and before there is full activation of the total skeletal muscle mass, and (iv) cardiovascular and other measures believed to relate to skeletal muscle anaerobiosis, including the maximum oxygen consumption (VO2 max) and the "anaerobic threshold", are indifferent predictors of exercise capacity in athletes with similar abilities. This review considers four additional models that need to be considered when factors limiting either short duration, maximal or prolonged submaximal exercise are evaluated. These additional models are: (i) the energy supply/energy depletion model; (ii) the muscle power/muscle recruitment model; (iii) the biomechanical model and (iv) the psychological model. By reviewing features of these models, this review provides a broad overview of the physiological, metabolic and biomechanical factors that may limit exercise performance under different exercise conditions. A more complete understanding of fatigue during exercise, and the relevance of the adaptations that develop with training, requires that the potential relevance of each model to fatigue under different conditions of exercise must be considered. PMID- 10843508 TI - Are social classes still relevant to analyse sports groupings in "postmodern" society? An analysis referring to P. Bourdieu's theory. AB - P. Bourdieu's theory will be used to analyse changes in sport sociology. A significant part of the transformation of sociological research depends on the analysis of social categories and particularly of social classes. The theory of P. Bourdieu grants social positions and conditions an important role in the explanation of an agent's behaviour. The question is to know whether sports groupings, in the so-called "postmodern" society, can still be analysed with Bourdieu's sociological frame of analysis. The observation of new forms of sports grouping is used, by some French sociologists, to criticise Bourdieu's axiomatic. It is argued here that it is more often a misunderstanding or a mechanical use of Bourdieu's theory which needs to be criticised. PMID- 10843509 TI - Rehabilitation following acute anterior cruciate ligament injuries--a 12-month follow-up of a randomized clinical trial. AB - The efficacy of two non-operative rehabilitation programs was studied in a consecutive randomized controlled clinical trial of 100 patients after 12 months subsequent to an acute anterior cruciate ligament (ACL) injury. Follow up of randomization to two training models was evaluated after 3 and 12 months: A self monitored training program (SM) of traditional mobility and muscle strength training of the injured leg was compared to a supervised (SV) training model exercising postural function in closed kinetic-chains. Nearly 50% of the patients in the SM group required supervision after 6 weeks. An intention-to-treat analysis was performed and showed significantly better values in most of the results of the supervised group at 3 and 12 months. An alternative analysis of subgroups showed a significant difference between transferred male patients and original SV male patients at 3 months but not at 12 months, indicating the importance of initial guiding after an ACL injury. No such difference was observed in the female patients. PMID- 10843510 TI - Ability of closed and open kinetic chain tests of muscular strength to assess functional performance. AB - The purpose of this study was to investigate the ability of closed and open kinetic chain tests of muscular strength to assess functional performance. Sixteen healthy male subjects, with a mean (+/-SD) age, body mass and height of 27+/-5 years, 78+/-9 kg and 183+/-9 cm, respectively, volunteered to participate in the study. In the closed kinetic chain test (involving muscles working across multiple joints), the subjects performed a 3 repetition maximum (3 RM) barbell squat. The open kinetic chain test (involving muscles working across a single joint) consisted of a concentric isokinetic knee extension at an angular velocity of 60 degrees/s, and was performed using a Kinetic Communicator II dynamometer. The test of functional performance (vertical jump) was performed with the subject standing erect, quickly performing a countermovement jump for maximal height. Moderately strong significant (P<0.05) correlations between the test of functional performance and the closed and open kinetic chain tests of muscular strength were noted, r=0.51 and r=0.57, respectively. It is suggested that the effect of training or rehabilitation interventions should not be based exclusively on tests of muscular strength. Rather, various forms of dynamometry including functional performance tests could be recommended. PMID- 10843511 TI - Aiming routines and their electrocortical concomitants among competitive rifle shooters. AB - The present study focused on an examination of competitive shooters' aiming process during a rifle shooting task. The barrel movements of the rifle, as detected by a laser system during the last 1000-ms time period preceding the triggering, were recorded from six elite and six pre-elite shooters. Electrocortical slow potentials (SPs) from frontal (Fz), centro-lateral (C3, C4), and occipital (Oz) brain areas were recorded to get an additional insight into the underlying covert processing. The results suggested that the elite shooters did not pull the trigger until they reached a sustained rifle position. In the pre-elite shooters the rifle appeared to be in a less stable position, and their strategy was to take advantage of the first appropriate moment of steadiness without a sustained rifle position so they could pull the trigger. The observed pre-trigger readiness potential (RP) shifts at Fz and Oz were more positive among the elite shooters relative to the pre-elite shooters, reflecting their more pronounced covert effort, rather than increasing preparedness for the trigger pull. The present study lends support for the view that a successful aiming strategy is mainly based on sustained rifle balancing. With regards to the brain slow potentials, it can be concluded that the RP shift does not specifically reflect the preparation for the trigger pull. PMID- 10843512 TI - Avulsion fracture of the iliac crest in a football player. AB - We describe an unusual injury of the iliac crest in an adolescent football player. The injury occurred as a result of a sudden twist of the trunk while kicking. Plain radiographs showed avulsion fracture of the anterior part of the iliac crest apophysis. Five months later the injury was partially ossificated but the patient felt minor pain and was not confident of returning to playing football. PMID- 10843513 TI - Six-month test-retest reliability of MRI-defined PET measures of regional cerebral glucose metabolic rate in selected subcortical structures. AB - Test-retest reliability of resting regional cerebral metabolic rate of glucose (rCMR) was examined in selected subcortical structures: the amygdala, hippocampus, thalamus, and anterior caudate nucleus. Findings from previous studies examining reliability of rCMR suggest that rCMR in small subcortical structures may be more variable than in larger cortical regions. We chose to study these subcortical regions because of their particular interest to our laboratory in its investigations of the neurocircuitry of emotion and depression. Twelve normal subjects (seven female, mean age = 32.42 years, range 21-48 years) underwent two FDG-PET scans separated by approximately 6 months (mean = 25 weeks, range 17-35 weeks). A region-of-interest approach with PET-MRI coregistration was used for analysis of rCMR reliability. Good test-retest reliability was found in the left amygdala, right and left hippocampus, right and left thalamus, and right and left anterior caudate nucleus. However, rCMR in the right amygdala did not show good test-retest reliability. The implications of these data and their import for studies that include a repeat-test design are considered. PMID- 10843514 TI - EEG recording during fMRI experiments: image quality. AB - Electroencephalographic (EEG) monitoring during functional magnetic resonance imaging (fMRI) experiments is increasingly applied for studying physiological and pathological brain function. However, the quality of the fMRI data can be significantly compromised by the EEG recording due to the magnetic susceptibility of the EEG electrode assemblies and electromagnetic noise emitted by the EEG recording equipment. We therefore investigated the effect of individual components of the EEG recording equipment on the quality of echo planar images. The artifact associated with each component was measured and compared to the minimum scalp-cortex distance measured in normal controls. The image noise originating from the EEG recording equipment was identified as coherent noise and could be eliminated by appropriate shielding of the EEG equipment. It was concluded that concurrent EEG and fMRI could be performed without compromising the image quality significantly if suitable equipment is used. The methods described and the results of this study should be useful to other researchers as a framework for testing of their own equipment and for the selection of appropriate equipment for EEG recording inside a MR scanner. PMID- 10843515 TI - Brain activation in the processing of Chinese characters and words: a functional MRI study. AB - Functional magnetic resonance imaging was used to identify the neural correlates of Chinese character and word reading. The Chinese stimuli were presented visually, one at a time. Subjects covertly generated a word that was semantically related to each stimulus. Three sorts of Chinese items were used: single characters having precise meanings, single characters having vague meanings, and two-character Chinese words. The results indicated that reading Chinese is characterized by extensive activity of the neural systems, with strong left lateralization of frontal (BAs 9 and 47) and temporal (BA 37) cortices and right lateralization of visual systems (BAs 17-19), parietal lobe (BA 3), and cerebellum. The location of peak activation in the left frontal regions coincided nearly completely both for vague- and precise-meaning characters as well as for two-character words, without dissociation in laterality patterns. In addition, left frontal activations were modulated by the ease of semantic retrieval. The present results constitute a challenge to the deeply ingrained belief that activations in reading single characters are right lateralized, whereas activations in reading two-character words are left lateralized. PMID- 10843516 TI - Tonotopic cortical changes following stapes substitution in otosclerotic patients: a magnetoencephalographic study. AB - The aim of the study was to investigate and follow up the tonotopic organization of the primary auditory cortex in otosclerotic patients before and after corrective surgery. The characteristics of primary auditory cortex activation were studied in ten otosclerotic patients (i.e., subjects suffering from a conductive hearing loss, prior to and following stapes substitution). Magnetoencephalographic recordings of auditory evoked fields by tone-burst stimulation at octave frequencies between 250 and 2000 Hz were performed during monaural stimulation. The brain topography of the main cortical response (N100m) generators at different tones was studied in patients and compared with ten healthy controls; pre- post-surgical changes were also correlated to their clinical outcome following corrective surgery. A significant decrease of the tonotopic extension in the cortical region responsive to the four explored frequencies was found in patients before surgery with respect to the control population. At the time of postsurgical follow-up, the tonotopic representation had enlarged and was approaching the dimensions seen in normal subjects, although with higher variability. The extent of the enlargement of the postoperative tonotopically organized area was directly correlated with the postsurgery period duration. Our findings indicate that auditory cortical areas of human adults undergo functional reorganization following peripheral alteration of the sensory input entering the CNS. The restriction of the cortical tonotopic region caused by the long-term reduction of acoustic input is followed by a reorganization within the usual boundaries following the recovery of auditory function; this process is taking place in a time scale of a few weeks. PMID- 10843517 TI - Voxel-based analysis of confounding effects of age and dementia severity on cerebral metabolism in Alzheimer's disease. AB - Alzheimer's disease is characterized by early hippocampal lesions, but neuropathological and functional imaging studies have also demonstrated involvement of associative cortices in patients suffering from this illness. New image-processing technologies have led to demonstration of predominant posteromedial cortical metabolic impairment in the disease. Confounding effects of both age and dementia severity on brain metabolism were assessed using categorical and correlational analyses performed with Statistical Parametric Mapping. Posterior cingulate and precuneus metabolism, assessed by positron emission tomography, was significantly correlated with age in a population of 46 patients with probable Alzheimer's disease. Metabolism in posterior cingulate and precuneus was higher in elderly than in younger patients with a diagnosis of Alzheimer's disease, even when dementia severity was taken as a confounding covariate. The data suggest that the sensitivity of positron emission tomography for the diagnosis of Alzheimer's disease is reduced in elderly cases, where less severe pathology is sufficient to induce clinical symptoms of dementia. Conversely, higher posteromedial metabolic impairment in early onset cases may reflect greater density of regional cerebral lesions or major decrease of functional afferences in a richly connected multimodal associative area. Posterior cingulate metabolism was also correlated to dementia severity, even when age was taken as a confounding covariate, whereas metabolism in the hippocampal formation was not shown to correlate with global cognitive deficit. Functional correlation was maintained between posterior cingulate and middle frontal cortex in demented patients as in elderly controls. The key role of posteromedial cortex in cognitive dysfunction assessed in Alzheimer's disease is probably related to its highly integrated position within attentional, visuospatial and memory neuronal networks. PMID- 10843518 TI - Multiobjective evolutionary algorithms: analyzing the state-of-the-art. AB - Solving optimization problems with multiple (often conflicting) objectives is, generally, a very difficult goal. Evolutionary algorithms (EAs) were initially extended and applied during the mid-eighties in an attempt to stochastically solve problems of this generic class. During the past decade, a variety, of multiobjective EA (MOEA) techniques have been proposed and applied to many scientific and engineering applications. Our discussion's intent is to rigorously define multiobjective optimization problems and certain related concepts, present an MOEA classification scheme, and evaluate the variety of contemporary MOEAs. Current MOEA theoretical developments are evaluated; specific topics addressed include fitness functions, Pareto ranking, niching, fitness sharing, mating restriction, and secondary populations. Since the development and application of MOEAs is a dynamic and rapidly growing activity, we focus on key analytical insights based upon critical MOEA evaluation of current research and applications. Recommended MOEA designs are presented, along with conclusions and recommendations for future work. PMID- 10843519 TI - Approximating the nondominated front using the Pareto Archived Evolution Strategy. AB - We introduce a simple evolution scheme for multiobjective optimization problems, called the Pareto Archived Evolution Strategy (PAES). We argue that PAES may represent the simplest possible nontrivial algorithm capable of generating diverse solutions in the Pareto optimal set. The algorithm, in its simplest form, is a (1 + 1) evolution strategy employing local search but using a reference archive of previously found solutions in order to identify the approximate dominance ranking of the current and candidate solution vectors. (1 + 1)-PAES is intended to be a baseline approach against which more involved methods may be compared. It may also serve well in some real-world applications when local search seems superior to or competitive with population-based methods. We introduce (1 + lambda) and (mu + lambda) variants of PAES as extensions to the basic algorithm. Six variants of PAES are compared to variants of the Niched Pareto Genetic Algorithm and the Nondominated Sorting Genetic Algorithm over a diverse suite of six test functions. Results are analyzed and presented using techniques that reduce the attainment surfaces generated from several optimization runs into a set of univariate distributions. This allows standard statistical analysis to be carried out for comparative purposes. Our results provide strong evidence that PAES performs consistently well on a range of multiobjective optimization tasks. PMID- 10843520 TI - Comparison of multiobjective evolutionary algorithms: empirical results. AB - In this paper, we provide a systematic comparison of various evolutionary approaches to multiobjective optimization using six carefully chosen test functions. Each test function involves a particular feature that is known to cause difficulty in the evolutionary optimization process, mainly in converging to the Pareto-optimal front (e.g., multimodality and deception). By investigating these different problem features separately, it is possible to predict the kind of problems to which a certain technique is or is not well suited. However, in contrast to what was suspected beforehand, the experimental results indicate a hierarchy of the algorithms under consideration. Furthermore, the emerging effects are evidence that the suggested test functions provide sufficient complexity to compare multiobjective optimizers. Finally, elitism is shown to be an important factor for improving evolutionary multiobjective search. PMID- 10843521 TI - Multiobjective satisfaction within an interactive evolutionary design environment. AB - The paper introduces the concept of an Interactive Evolutionary Design System (IEDS) that supports the engineering designer during the conceptual/preliminary stages of the design process. Requirement during these early stages relates primarily to design search and exploration across a poorly defined space as the designer's knowledge base concerning the problem area develops. Multiobjective satisfaction plays a major role, and objectives are likely to be ill-defined and their relative importance uncertain. Interactive evolutionary search and exploration provides information to the design team that contributes directly to their overall understanding of the problem domain in terms of relevant objectives, constraints, and variable ranges. This paper describes the development of certain elements within an interactive evolutionary conceptual design environment that allows off-line processing of such information leading to a redefinition of the design space. Such redefinition may refer to the inclusion or removal of objectives, changes concerning their relative importance, or the reduction of variable ranges as a better understanding of objective sensitivity is established. The emphasis, therefore, moves from a multiobjective optimization over a preset number of generations to a relatively continuous interactive evolutionary search that results in the optimal definition of both the variable and objective space relating to the design problem at hand. The paper describes those elements of the IEDS relating to such multiobjective information gathering and subsequent design space redefinition. PMID- 10843522 TI - Efficient and scalable Pareto optimization by evolutionary local selection algorithms. AB - Local selection is a simple selection scheme in evolutionary computation. Individual fitnesses are accumulated over time and compared to a fixed threshold, rather than to each other, to decide who gets to reproduce. Local selection, coupled with fitness functions stemming from the consumption of finite shared environmental resources, maintains diversity in a way similar to fitness sharing. However, it is more efficient than fitness sharing and lends itself to parallel implementations for distributed tasks. While local selection is not prone to premature convergence, it applies minimal selection pressure to the population. Local selection is, therefore, particularly suited to Pareto optimization or problem classes where diverse solutions must be covered. This paper introduces ELSA, an evolutionary algorithm employing local selection and outlines three experiments in which ELSA is applied to multiobjective problems: a multimodal graph search problem, and two Pareto optimization problems. In all these experiments, ELSA significantly outperforms other well-known evolutionary algorithms. The paper also discusses scalability, parameter dependence, and the potential distributed applications of the algorithm. PMID- 10843523 TI - Phenotypic assays and sequencing are less sensitive than point mutation assays for detection of resistance in mixed HIV-1 genotypic populations. AB - The sensitivity and discriminatory power of the 151 and 215 amplification refractory mutation system (ARMS) were evaluated, and their performance for the detection of drug resistance in mixed genotypic populations of the reverse transcription (RT) gene of HIV-1 were compared with T7 sequencing, cycle sequencing, the line probe assay (LiPA) HIV-1 RT test, and the recombinant virus assay (RVA). ARMS and the LiPA HIV-1 RT test were shown to be able to detect minor variants that in particular cases comprised only 1%. T7 sequencing on an ALF semiautomated sequencer could correctly score mixtures only when variants were present at 50%. Cycle sequencing on an ABI PRISM 310 improved the sensitivity for mixtures to about 25%. Using RVA, it was shown that at least 50% of the virus population needed to carry the resistance mutation at codon 184 to afford phenotypic resistance against lamivudine. The two point mutation assays therefore proved to be more sensitive methods than sequencing and RVA to reliably determine a gradual shift in HIV-1 drug resistance mutations in follow-up of patients infected with HIV-1. In 4 of 5 treated patients who were followed by ARMS, a gradual shift in resistant genotypic populations was observed during a period of 6 to 19 months. For 1 patient, a shift from wild to mutant type at position 151 occurred within 2 months, without mixed genotypic intermediate type's being detected. PMID- 10843524 TI - Survival and failure to thrive in the SIV-infected juvenile rhesus monkey. AB - In AIDS patients, wasting in adults and failure to thrive in children are common and devastating problems. Weight loss in rhesus macaques infected with simian immunodeficiency virus (SIV) has not been well characterized. The purpose of this study was to determine growth curves in SIV-infected juvenile macaques to determine the effects of SIV infection on body weight and growth. Medical records of seven juvenile male SIV-infected macaques were retrospectively reviewed to determine body weights, survival time, CD4 count, and viral load. Mean age and body weight at the time of inoculation were 63.3 weeks and 2.4 kg, respectively. Mean survival was 73.7 weeks, and mean body weight at the time of death was 3.0 kg, whereas the published mean body weight for this age of male rhesus macaque is 4.1 kg. Compared with the linear growth pattern of normal animals, the growth pattern for the SIV-infected animals exhibited strong nonlinearity with an inflection point at the mean survival of 74 weeks. After this time point, the discrepancy between growth curves for infected and healthy animals increased at a greater rate. Body weight correlated inversely with viral load (r = -0.368; p = .003) but there was no correlation between body weight and CD4 count. The results of this study suggest that failure to thrive is a consequence of SIV infection and may be related to severity of infection. PMID- 10843525 TI - HIV-1 RNA in plasma and genital tract secretions in women infected with HIV-1. AB - To assess antiretroviral therapy, all compartments, including the genital tract, need to be evaluated. HIV-1 RNA was quantified in whole cervicovaginal lavage fluid (CVL) and plasma of 56 women and in the cellular and supernatant fractions of 27 of these women. Overall, we detected HIV-1 RNA in 59% of whole CVL samples and in 61% and 44% of cellular and supernatant fractions of the subset of women, respectively. Detectability of HIV-1 RNA in CVL increased with increasing level of plasma RNA in both unfractionated and cell-associated CVL components (p = .0004 and .002, respectively), but not in the cell-free fraction (p = .29). Mean HIV-1 RNA levels in CVL increased with decreasing CD4 counts (p = .002,) and with increasing plasma HIV-1 RNA (p < .001). Adjusted odds ratios (OR) for detectable CVL RNA were highest for women with CD4 counts <200 cells/mm3 (OR, 10.1; 95% confidence interval [CI]: 1.6-82.7; p = .02) and >50,000 copies/ml of plasma RNA (OR, 25.2; 95% CI, 3.2-554; p = .01). Treatment did not seem to affect RNA detection in CVL after adjusting for plasma RNA and CD4. In conclusion, we found that detectability and level of CVL RNA were closely associated with the cellular fraction of genital secretions in women and strongly correlated with the level of plasma RNA and CD4. Genital tract secretions may need to be tested in the assessment of treatment efficacy and this can easily be accomplished with this rapid and easy procedure using whole CVL. PMID- 10843527 TI - A novel genotype encoding a single amino acid insertion and five other substitutions between residues 64 and 74 of the HIV-1 reverse transcriptase confers high-level cross-resistance to nucleoside reverse transcriptase inhibitors. Abacavir CNA2007 International Study Group. AB - We investigated HIV-1 reverse transcriptase (RT) polymorphisms of plasma isolates from 98 HIV-1-infected study subjects with >2 years of antiretroviral therapy who were failing their current protease inhibitor (PI)-containing regimen. In 1 patient, we detected a virus with a heavily mutated beta3-beta4 connecting loop of the HIV-1 RT fingers subdomain, consisting of a single aspartate codon insertion between positions 69 and 70 and five additional variations: 64N, K65, K66, 67G, 68Y, T69, Ins D, 70R, W71, R72, K73, 74I. Mutants with the recently described 2-aa insertions between codons 68 and 70 of RT were detected in another 3 patients. Among the four isolates with the 1- or 2-aa insertions, the novel genotype was the most refractory to therapy and displayed the highest level of phenotypic resistance to nucleoside reverse transcriptase inhibitors (NRTIs). Follow-up samples demonstrated that the novel mutant represents a stable genetic rearrangement and that the amino acid insertions can coexist with nonnucleoside analogue reverse transcriptase inhibitors (NNRTI) mutations resulting in phenotypic resistance to both NRTIs and NNRTIs. An increasing number of HIV-1 isolates containing various insertions in the beta3-beta4 hairpin of the HIV-1 RT fingers subdomain appear to emerge after prolonged therapy with different NRTIs, and these polymorphisms can confer multiple drug resistance against NRTIs. PMID- 10843526 TI - High-dose saquinavir plus ritonavir: long-term efficacy in HIV-positive protease inhibitor-experienced patients and predictors of virologic response. AB - The year-long antiviral efficacy of a high-dose salvage regimen consisting of saquinavir (800 mg twice daily) plus ritonavir (400 mg twice daily) was evaluated in 58 HIV-positive patients who had seen no improvement under first-line protease inhibitor-containing regimens, nor in baseline predictors of virologic response. The efficacy of therapy was determined by CD4+/CD8+ and HIV-1 RNA values. The primary endpoint of our study was the percentage of patients with HIV-1 RNA levels <200 copies/ml (virologic success) at 6 and 12 months of of follow-up. Secondary endpoints were log10 reduction in HIV-1 RNA levels and CD4+ increases through follow-up. Surrogate markers related with a lower HIV-1 RNA area under the curve were identified at baseline. Kaplan-Meier analysis and Cox proportional hazards models were applied to identify baseline predictors of achieving viral suppression at <200 copies/ml. All analyses were intention to treat-last observation carried forward. Patients achieved a median HIV-1 RNA level reduction of >0.5 log through 1 year (-0.59 log10 at 12 months), as well as CD4+ counts increased significantly (89 cells/mm3 at 12 months). Overall, 53% of patients were likely to achieve HIV-1 RNA levels <200 copies/ml at 6 months. Seventy-six percent of patients who started therapy at HIV-1 RNA levels <5000 copies/ml but only 42% with baseline viral load of 5000 to 30,000 copies/ml and 18.7% with baseline viral load >30,000 copies/ml were likely to achieve viral suppression at 6 months (p < .001, log-rank test). Patients with baseline HIV-1 RNA levels between 5000 and 30,000 copies/ml (relative hazard [RH], 0.39; 95% confidence interval [CI], 0.01 to 0.98; p = .0396) and patients with baseline HIV-1 RNA levels >30,000 copies/ml (RH, 0.20; 95% CI, 0.07-0.61; p = .0040) were less likely to reach undetectable HIV-1 RNA levels than those with baseline HIV-1 RNA levels <5000 copies/ml. Salvage highly active antiretroviral therapy (HAART) strategies including saquinavir (SQV) at high doses plus ritonavir (RTV) exert a significant long-term efficacy in more than half of PI-experienced patients without significant additional toxicity. This therapeutic efficacy is strongly implemented by a switch at the lower HIV-1 RNA levels. PMID- 10843528 TI - Dehydroepiandrosterone sulfate (DHEAS) and testosterone: relation to HIV illness stage and progression over one year. AB - This study explored associations between serum dehydroepiandrosterone sulfate (DHEAS), free and total testosterone levels, and HIV illness markers, including viral load, and the behavioral problems of fatigue and depressed mood. Subjects were 169 HIV-positive men evaluated at baseline, 6, and 12 months for levels of DHEAS, total and free testosterone, HIV RNA, CD4, HIV symptoms, opportunistic illnesses, fatigue, and depression. Men with AIDS (N = 105), compared with men with less advanced illness, had lower mean levels of DHEAS. Baseline DHEAS was positively correlated with CD4 count, HIV symptom severity, and was inversely correlated with HIV RNA. Baseline DHEAS below the laboratory reference range (96 microg/dl) was associated with history of opportunistic infections and malignancies (adjusted odds ratio [OR], 4.4; 95% confidence interval [CI], 1.9 10.4) and with incidence of these complications or death over 1 year (adjusted OR, 2.6; 95% CI, 1-7.2). Initiating protease inhibitor combination therapy was associated with an increase in DHEAS over 6 months. Free testosterone was inversely correlated with HIV RNA, but there were no other significant associations between testosterone and HIV illness markers. No hormone was related to fatigue or depression. This study confirms that low serum DHEAS is associated with HIV illness markers, including viral load, and carries negative prognostic value. Further, protease inhibitor therapy may result in increased circulating DHEAS. PMID- 10843529 TI - Bacteremia in HIV-infected patients: short-term predictors of mortality. AB - To identify characteristics associated with mortality in HIV-infected patients with bacteremia, 88 bacteremic episodes in 80 HIV-infected patients were prospectively identified over a 5-month period and observed for 30 days. Demographic, clinical, laboratory, and radiologic data were collected. Mean and median age was 41 years. Most study subjects were homosexual men. Median CD4 count was 20 cells/mm3. Gram-positive organisms predominated (65%). The most common source of bacteremia was intravascular catheters (45%). Overall mortality was 30%. A history of malignancy, three or more opportunistic infections, shock, low hemoglobin, source of bacteremia other than an intravascular catheter, resistance to therapy, and a second bacteremic episode during the study period, were all found to be independent predictors of mortality. In this cohort of HIV infected patients, most of whom were severely immunosuppressed, several factors were found to be significantly and independently associated with mortality. PMID- 10843530 TI - Prevalence and incidence of HIV among incarcerated and reincarcerated women in Rhode Island. AB - This study explores recent temporal trends in HIV prevalence among women entering prison and the incidence and associated risk factors among women reincarcerated in Rhode Island. Results from mandatory HIV testing from 1992 to 1996 for all incarcerated women were examined. In addition, a case control study was conducted on all seroconverters from 1989 to 1997. In all, 5836 HIV tests were performed on incarceration in 3146 women, 105 of whom tested positive (prevalence, 3.3%). Between 1992 and 1996, the annual prevalence of HIV among all women known to be HIV-positive was stable (p = .12). Age >25 years, nonwhite race, and prior incarceration were associated with seropositivity. Of 1081 initially seronegative women who were retested on reincarceration, 12 seroconverted during 1885 person years (PY) of follow-up (incidence, 0.6/100 PY). Self-reported injection drug use (IDU; odds ratio [OR], 3.7; 95% confidence interval [CI], 1.3-10.1) was significantly associated with seroconversion, but sexual risk was not (OR, 1.1; 95% CI, 0.4-3.5). Incarceration serves as an opportunity for initiation of treatment and linkage to community services for a population that is at high risk for HIV infection. PMID- 10843531 TI - HIV-1 RNA levels in an African population-based cohort and their relation to CD4 lymphocyte counts and World Health Organization clinical staging. AB - Apart from a small number of reports from people who are based in hospitals, data on viral load in HIV-infected people in sub-Saharan Africa, where most infections occur, are lacking. We report serum HIV-1 RNA levels in a population-based cohort in rural Uganda using the nucleic acid sequence-based amplification procedure (NASBA) test kit and describe their relation to CD4 counts and World Health Organization (WHO) clinical staging. The median (interquartile range [IQR]) viral loads were 87,000 copies/ml (37,500-295,000 copies/ml) in 40 prevalent cases infected for >6 years, and 31,000 copies/ml (7800-174,000 copies/ml) in 65 incident cases with seroconversion dates within the previous 6 years. Although we found a correlation between viral load and absolute CD4 count (p < .0001), there was no evidence for an association with CD4 decline (p = .1). Overall, there was a significant trend of increasing viral load with worsening clinical stage from a median viral load of 15,000 for those in WHO stage 1 (asymptomatic) to 150,600 copies/ml for those in stage 4 (AIDS; p < .001). However, the association was seen only in incident cases. Thus, we found that the NASBA test on serum was a useful indicator of disease stage especially in persons known to be infected for <6 years. Such baseline data are important for vaccine research, and if antiretroviral drugs become available to more than a few people in Africa, it will be important that accurate viral load estimations are available at least in a proportion of people to monitor the effectiveness of treatment, and measure the compliance and emerging resistance to these drugs. PMID- 10843532 TI - High frequency of non-B subtypes in newly diagnosed HIV-1 infections in Switzerland. AB - HIV-1 subtypes were determined in newly diagnosed residents of Switzerland. Blood was anonymously collected from patients with a first confirmed positive HIV-1 test result. Viral DNA from the env V3-V5 region was amplified by nested polymerase chain reaction (PCR) and screened for subtype B by heteroduplex mobility assay. All amplicons not identified as B were sequenced. From November 1996 to February 1998, 206 samples were analyzed. Main transmission risks were unprotected heterosexual (55.7%) or homosexual (27.1%) sexual contact or intravenous drug use (12.9%). Subtype B dominated in patients of Swiss, other European, American, or Asian citizenship; particularly high frequencies were found in homosexuals (97%) and drug users (94%). Non-B subtypes including A, C, D, E, F, G, H, a possible B/F recombinant, and a sequence related to J were present in 28.2% (95% confidence interval [CI], 22.9%-35.0%). Non-B were frequent in African citizens (95%), heterosexually infected individuals (44%), and women (43%). Heterosexually infected Swiss males harbored non-B strains in 18% and females in 33%. The results document a change in the epidemiology of newly diagnosed HIV-1 infections in Switzerland: predominance of heterosexual transmission and a high frequency of non-B subtypes. PMID- 10843534 TI - Predictive markers of HIV-related weight loss and determination of differences between populations with weight loss stratified by opportunistic processes. AB - OBJECTIVE: To describe factors predictive of >10% weight loss among enrolled participants in clinical trials of the AIDS Clinical Trial Group (ACTG). DESIGN: A retrospective analysis of data from selected ACTG antiretroviral clinical trials completed prior to 1996 (ACTG 116, 117, 155, 175, and 241), which did not include protease inhibitors. METHODS: Data were analyzed in Cox proportional hazards models to determine significant predictors for >10% weight loss while on study. Weight loss occurring within 30 days before or after an opportunistic infection (OI) was defined as "OI-associated." Both univariate and multivariate models were considered; gender-specific models were also analyzed to provide insight into potential gender differences in predictors of weight loss. RESULTS: We found that substantial weight loss is a frequent occurrence among those enrolled in clinical trials of antiretroviral agents; approximately 15% of subjects in the studies considered experienced >10% weight loss. CD4 cell count and HIV-1 RNA at week 8, Karnofsky score, and injection drug use status were significant multivariate predictive markers for weight loss associated with an OI; baseline weight, hemoglobin, triglycerides, and gender were additional predictors for weight loss not associated with an OI. CONCLUSIONS: This is the first study to characterize the association between baseline viral load and future weight loss. Baseline and week 8 immunologic parameters as well as measures of baseline symptomatology were significant predictors of weight loss associated and not associated with an OI. PMID- 10843533 TI - Emerging HIV infections with distinct subtypes of HIV-1 infection among injection drug users from geographically separate locations in Guangxi Province, China. AB - Heroin users from Guangxi province, a southern province of China that borders Vietnam in the south and Yunnan province in China in the west, were studied for prevalence and risk factors for HIV-1 infection. Viral env sequences from HIV-1 positive individuals were also determined for subtypes of HIV-1. The overall HIV prevalence among 227 heroin users was 40%. Most had used drugs for < or = 3 years. Sharing of injection equipment and unprotected sex were significantly associated with HIV-1 infection. Subtypes C and E HIV-1 were detected in infected heroin users and were sharply segregated in two geographic locations: only subtype C was found in a border city with Yunnan province, whereas only subtype E was found in a city bordering northern Vietnam. HIV-1 strains within each subtype were remarkably homogenous, with a mean intersubject DNA distance of 2.32% for subtype E and 1.13% for subtype C, respectively. Phylogenetic analysis of C2-V5 region of Guangxi subtype E env sequences revealed significant clustering with subtype E sequences from southern Vietnam and Cambodia. These results suggest that HIV-1 infection among heroin users in Guangxi represents two emerging epidemics initiated from distinct sources: one from Vietnam and another from Yunnan province. Factors associated with HIV-1 infection were not restricted to injection practices. Unprotected sexual behaviors are likely to increase the probability of HIV transmission beyond this high-risk population. Designing and implementing effective intervention strategies targeted toward both injection drug use and high risk sexual behavior are urgently needed to further reduce HIV 1 spread in China. PMID- 10843535 TI - HIV-1 transmission in injection paraphernalia: heating drug solutions may inactivate HIV-1. AB - In response to recent concerns about risk of HIV-1 transmission from drug injection paraphernalia such as cookers, ethnographic methods were used to develop a descriptive typology of the paraphernalia and practices used to prepare and inject illegal drugs. Observational data were then applied in laboratory studies in which a quantitative HIV-1 microculture assay was used to measure the recovery of infectious HIV-1 in cookers. HIV-1 survival inside cookers was a function of the temperature achieved during preparation of drug solutions; HIV-1 was inactivated once temperature exceeded, on average, 65 degrees C. Although different types of cookers, volumes, and heat sources affected survival times, heating cookers 15 seconds or longer reduced viable HIV-1 below detectable levels. PMID- 10843536 TI - Molecular pathologic analysis of the tonsil in HTLV-I-infected individuals. AB - Little is known about the role of the tonsils in HTLV-I infection. We performed molecular pathologic studies of tonsils in individuals positive or negative for anti-HTLV-I antibodies (HTLV-I-Ab) to clarify histologic characteristics of tonsils in HTLV-I infection. We collected tonsils and peripheral blood samples from patients who underwent tonsillectomy in a prospective manner. HTLV-I-Ab in serum was examined and presence of HTLV-I provirus was detected by polymerase chain reaction (PCR) in extracted DNA of both peripheral blood and tonsils. Histopathologic and immunohistochemical evaluations of tonsils were performed. HTLV-I seropositivity and PCR detection of HTLV-I provirus matched perfectly. Tonsil samples from seropositive individuals showed atrophy of the mantle zone and high numbers of T cells in the marginal zone compared with findings in HTLV-I negative samples. HTLV-I provirus could be detected only from extracted DNA of extrafollicular areas. PCR in situ hybridization also showed positive signals in some mononuclear cells located in the marginal zone. There was a significant correlation between HTLV-I proviral load in tonsils and in peripheral blood. These results suggest the presence of characteristic histologic changes and deviated localization of HTLV-I-infected cells in the tonsils of individuals positive for HTLV-I. PMID- 10843538 TI - Development of Kaposi's sarcoma despite sustained suppression of HIV plasma viremia. PMID- 10843537 TI - Plasma RNA viral load as measured by the branched DNA and nucleic acid sequence based amplification assays of HIV-1 subtypes A and D in Uganda. PMID- 10843539 TI - Mother-to-child HIV transmission. European Study Group on the Natural History of HIV Infection in Women. PMID- 10843540 TI - Using a differential refractive index detector as a pressure transducer for online viscometry in exclusion chromatography. AB - A differential refractive index (DRI) detector was tested as a pressure transducer for single capillary online viscometry in exclusion chromatography. The relationship between the detector response and pressure is linear, in agreement with the theoretically expected influence of pressure on refractive index and its dependence on temperature is negligible. Whole polymer and local intrinsic viscosities were determined and compared for narrow and, respectively, broad molecular weight distribution polymer samples. Considering that the detection system described can be improved, the results suggest that modern DRI detectors are susceptible to satisfy the requirements for a suitable pressure transducer in this application. PMID- 10843541 TI - Pellicular expanded bed matrix suitable for high flow rates. AB - A new type of expanded bed matrix with a heavy core of stainless steel covered with an agarose layer was prepared. Two bead size fractions, the smaller one (32 75 microm diameter) having a single particle core and the larger (75-180 microm diameter) with an agglomerate of stainless steel particles constituting the core, were chosen for further characterisation. The dispersion behaviour was determined both in packed bed and expanded bed modes by the retention time distribution method (RTD) and compared with the Streamline matrix (Amersham Pharmacia Biotech). The comparison turned out in favour of the new matrix. Flow rates as high as 3000 cm/h were used with the larger fraction, giving stable expanded beds with good mass transfer properties. The matrices were mechanically stable without any tendency to crack or peal, even after prolonged use. PMID- 10843542 TI - Multiple solid-phase microextraction. AB - Theoretical aspects of multiple solid-phase microextraction are described and the principle is illustrated with the extraction of lidocaine from aqueous solutions. With multiple extraction under non-equilibrium conditions considerably less time is required in order to obtain an extraction yield that is equal to that of one extraction at equilibrium. On the other side, the extraction yield can be increased if multiple extraction is performed with the same total time as is needed for one extraction at equilibrium time. The effect of multiple extraction is strongly dependent on the value of the partition constant and for practical use the length of the desorption time is important. A good agreement between theoretical and experimental data has been obtained. Chromatograms are presented showing the potential of multiple solid-phase microextraction. PMID- 10843543 TI - Case of enantiomer impurity identification by normal-phase chiral high performance liquid chromatography with optical rotation and mass spectrometric detection. AB - An impurity produced in the synthesis of compound I is separated and identified as its enantiomer II using normal-phase chiral high-performance liquid chromatography (HPLC) with UV absorbance, optical rotation (OR) and mass spectrometric (MS) detection. The results show that the impurity II and compound I have equal and opposite specific rotations, identical MS spectra and the same MS-MS fragmentation pattern, as required for enantiomers. The procedures presented demonstrate a novel combination of methods for enantiomer identification and characterization that do not require the preparation of individual enantiomer markers or even the racemic mixture, thus reducing the need for additional synthetic work. PMID- 10843544 TI - Analysis of linear and cyclic oligomers in polyamide-6 without sample preparation by liquid chromatography using the sandwich injection method. II. Methods of detection and quantification and overall long-term performance. AB - By separating the first six linear and cyclic oligomers of polyamide-6 on a reversed-phase high-performance liquid chromatographic system after sandwich injection, quantitative determination of these oligomers becomes feasible. Low wavelength UV detection of the different oligomers and selective post-column reaction detection of the linear oligomers with o-phthalic dicarboxaldehyde (OPA) and 3-mercaptopropionic acid (3-MPA) are discussed. A general methodology for quantification of oligomers in polymers was developed. It is demonstrated that the empirically determined group-equivalent absorption coefficients and quench factors are a convenient way of quantifying linear and cyclic oligomers of nylon 6. The overall long-term performance of the method was studied by monitoring a reference sample and the calibration factors of the linear and cyclic oligomers. PMID- 10843545 TI - Amperometric determination of underivatized amino acids at a nickel-modified gold electrode by anion-exchange chromatography. AB - A nickel-based composite electrode obtained by anodic electrodeposition of nickel (III) oxyhydroxide film on the gold electrode substrate was characterized as an amperometric sensor and successfully applied to the determination of underivatised amino acids in flow-through systems. The electrodeposition of nickel oxyhydroxide films was obtained by cycling a gold electrode between 0.0 V and +1.0 V vs. a saturated calomel electrode in a 80 microM Ni2+ solution buffered at pH 10 with NaHCO3/Na2CO3. The resulting Au-Ni composite electrode exhibits good stability in alkaline medium and can be used as an amperometric sensor of underivatised amino acids at a fixed applied potential (+0.55 V vs. Ag/AgCl). The detection limits (S/N=3) for all investigated compounds ranged between 5 and 30 pmol injected, while the linear ranges spanned over two or three orders of magnitude. The contents of several free amino acids in two sample cheeses from different brands were evaluated by calibration graphs. PMID- 10843546 TI - Determination of butyltin and phenyltin species by reversed-phase liquid chromatography and fluorimetric detection. AB - Chromatographic separation of monobutyltin (MBT), monophenyltin (MPhT), dibutyltin (DBT), diphenyltin (DPhT), tributyltin (TBT) and triphenyltin (TPhT) was studied using end-capped reversed-phases (RP) and methanol-acetic acid-water or acetronitrile-acetic acid-water mixtures as mobile phases. Several RP columns were evaluated, and the effect of acetic acid, oxalic acid, triethylamine, and organic modifier on peak shape and retention was examined. A method based on gradient elution RPLC and fluorimetric detection is proposed for the determination of DBT, DPhT, TBT and TPhT. The sensitivity of the method makes it suitable for environmental analysis. PMID- 10843547 TI - Determination of the plant growth regulator chlormequat in food by liquid chromatography-electrospray ionisation tandem mass spectrometry. AB - A confirmatory method for the determination of trace levels of chlormequat in a variety of different food matrices was developed. It entails a single clean-up step over a solid-phase cation exchange resin and subsequent liquid chromatography-electrospray ionisation tandem mass spectrometry using a stable isotopically labelled internal standard. Mass spectral acquisition was done in selected reaction monitoring mode, selecting the transitions from both the 35Cl and the 37Cl isotope of chlormequat. Recoveries after extraction and clean-up, determined with radio-labelled chlormequat and averaged over the spiking range (16-65 microg kg(-1)) in four different commodities, were within 88-96%, with a coefficient of variation better than 8%. The method can be applied to pears, pear juice concentrates, fruit purees, and cereal products, with typical limits of detection for chlormequat estimated at 2-5 microg kg(-1). A survey of different food commodities revealed that chlormequat was detectable--albeit at very low levels--in many of the food samples analysed, with the highest concentration recorded in pears purchased in Switzerland and of South African origin (5.5 mg kg(-1)). Measurements were also conducted on two LC-MS instruments and demonstrate the versatility and robustness of the method and its applicability to instruments of different ion source design. PMID- 10843548 TI - Simultaneous determination of acidic and non-acidic pesticides in natural waters by liquid chromatography-mass spectrometry. AB - There is increasing interest and demand for real multi-residue methods able to simultaneously determine pesticides with a broad spectrum of chemical characteristics in environmental and biological matrices. A method based on solid phase extraction with a Carbograph 4 cartridge and liquid chromatography with electrospray mass spectrometry (LC-ES-MS) enabling simultaneous determination of non-acidic and acidic pesticides in real water samples is described. On repeatedly (n=5) extracting 4 l of drinking water (spike level 50 ng/l), 2 l of ground water (spike level 100 ng/l) and 1 l of river water (spike level 200 ng/l), recovery of 26 base/neutral pesticides and 13 acidic pesticides were equal to or better than 80%, except for carbendazim (67%), butocarboxim (73%), aldicarb (75%) and molinate (77%). Relative standard deviations ranged between 4 and 15%. Final extracts containing acidic and non-acidic pesticides were analyzed in a single chromatographic run while the ES-MS system was operated in both positive and negative ion modes. With the aim of finding the best operating conditions, in terms of sensitivity, the pH of the LC eluent was varied in the 2.9-8.4 range. Altogether, the best results were obtained by using an LC eluent containing 1 mmol/l formic acid. Over the entire pH range considered, well shaped peaks for both basic and acidic analytes were achieved by the use of a new generation LC column. By extracting selected ion current profiles from the total ion current mass chromatogram relative to analysis of 4 l of drinking water spiked with 50 ng/l of each of the 39 analytes, estimated limits of detection ranged between 0.05 and 1.5 ng/l, except for propyzamide (8 ng/l) and 2,4-DB (3 ng/l). PMID- 10843549 TI - Simultaneous quantitative trace analysis of anionic and nonionic surfactant mixtures by reversed-phase liquid chromatography. AB - The aim of this work was to simultaneously analyse mixtures of a polydisperse polyethylene oxide (PEO) nonionic surfactant and an anionic surfactant (sodium dodecylsulphate, SDS) in water containing sodium chloride in order to quantify trace amounts of these mixtures after their adsorption at water-solid interfaces. A fractional factorial design was then used to optimise the separation by ion pair reversed-phase liquid chromatography as a function of six factors: the chain length of the tetraalkylammonium salt used as ion-pairing reagent which varied from methyl (C1) to n-propyl (C3); the concentration of this ion-pairing salt; the acetonitrile percentage in water used as organic modifier; the flow-rate; the temperature of analysis and also the sodium chloride concentration. The factorial design enabled in a limited number of analyses, not only to determine which factors had significant effects on retention times or on resolution between a pair of nonionic oligomers, but also to modelize and then find the interesting and rugged area where this resolution was optimal as well as the conditions where time of analysis was not prohibitive. After optimisation of HPLC analysis, we used a trace enrichment procedure to quantify very low concentrations of SDS and C12E9 polydisperse PEO in water. A C18 cartridge and a strong anionic exchange cartridge were coupled and the conditions of elution were optimised in order to obtain concentrated samples which were injected in the same eluent than the HPLC mobile phase. Under such conditions, we were able to quantify, in a single run, mixtures of anionic and nonionic surfactants at concentrations as low as 3.6 microg l(-1) for SDS and 2.5 microg l(-1) for each PEO oligomer in water. PMID- 10843550 TI - Revised linear solvation energy relationship coefficients for the 77-phase McReynolds data set based on an updated set of solute descriptors. AB - Linear solvation energy relationship (LSER) coefficients for the 77-phase McReynolds data set have been recalculated using updated solute descriptors in the revised solvation equation: [equation: see text] These revised LSER coefficients are presented and classified by cluster analysis into groupings of stationary phases which have comparable solubility properties. It was found that the groupings were similar to those proposed by Abraham using the original solvation equation and that any dissimilarities were readily explainable by the grouping methods that were applied. Comparison of the original coefficients with the revised set also shows that several stationary phases which had a statistically insignificant b1 value with the original equation now have significant b1 values when utilizing the revised solvation equation. PMID- 10843551 TI - Hold-up time in gas chromatography. V. Dependence of the retention of n-alkanes on the chromatographic variables in isothermal gas chromatography. AB - The chromatographic behaviour of n-alkanes and other homologous series in isothermal gas chromatography has been shown to depart from the "linear" representation of the logarithms of the adjusted retention times vs. carbon number. One of the expressions proposed to describe this behaviour is tR(z)=A+exp(B+CzD). In this paper, a regression analysis shows that three of the parameters of the equation depend on different chromatographic variables such as hold-up time, average linear gas velocity, volume and polarity of the stationary phase and temperature of the column. The fourth parameter (D), responsible for the departure from the "linearity", does not depend on any chromatographic variable, and represents a gradual decrease of the contribution of the methylene groups to the general properties of n-alkanes, with no relation to the chromatographic phenomenon. PMID- 10843552 TI - Capillary zone electrophoretic separation of neutral species of chloro-s triazines in the presence of cationic surfactant monomers. AB - Chloro-s-triazines are difficult to separate by capillary zone electrophoresis (CZE), due to their low pKa values. However, these analytes can be effectively separated by CZE in the presence of cationic surfactant monomers, such as tetradecylammonium bromide (TTAB) and dodecyltrimethylammonium bromide (DTAB). The separation mechanism based on a 1:1 binding of analytes to cationic surfactant monomers is proposed. The binding constants of chloro-s-triazines to cationic surfactant monomers are estimated. The results show that the strength of the interactions of these analytes with TTAB monomers is considerably strong, whereas that of the corresponding analyte with DTAB monomers is about 12- to 14 fold weaker. A linear correlation of binding constants with log P(ow) (the logarithm of the partition coefficient of analytes between 1-octanol and aqueous phases) indicates that the migration order of these chloro-s-triazines depends primarily on their hydrophobicity. Moreover, the skewed peaks of chloro-s triazines observed may reveal the occurrence of adsolubilization of these analytes in the adsorbed cationic surfactant layer on the capillary surface. PMID- 10843553 TI - Analysis of tetramethylpyrazine in Ephedrae herba by gas chromatography-mass spectrometry and high-performance liquid chromatography. AB - A simple and reliable HPLC method was developed for the determination of 2,3,5,6 tetramethylpyrazine (TMP) in Ephedrae herba. Further identification of TMP was achieved using GC-MS. The mobile phase used was methanol-water-35% acetic acid (35:65:0.5, v/v/v) at a flow-rate of 0.8 ml/min. The detection wavelength was set at 290 nm. The linear range of the peak area calibration curve of TMP was 2.64 264 mg/l (r=0.9987) and the recovery for TMP in Ephedrae herba extracts was 101.1 106.9%. The relative standard deviations of retention time and peak area were 0.18 and 1.5% (n=6), respectively. The detection limit of TMP was 0.03 mg/l. The contents of TMP in Ephedrae herba could easily be determined within 10 min. PMID- 10843554 TI - Genetic diversity and epidemiology of infectious hematopoietic necrosis virus in Alaska. AB - Forty-two infectious hematopoietic necrosis virus (IHNV) isolates from Alaska were analyzed using the ribonuclease protection assay (RPA) and nucleotide sequencing. RPA analyses, utilizing 4 probes, N5, N3 (N gene), GF (G gene), and NV (NV gene), determined that the haplotypes of all 3 genes demonstrated a consistent spatial pattern. Virus isolates belonging to the most common haplotype groups were distributed throughout Alaska, whereas isolates in small haplotype groups were obtained from only 1 site (hatchery, lake, etc.). The temporal pattern of the GF haplotypes suggested a 'genetic acclimation' of the G gene, possibly due to positive selection on the glycoprotein. A pairwise comparison of the sequence data determined that the maximum nucleotide diversity of the isolates was 2.75% (10 mismatches) for the NV gene, and 1.99% (6 mismatches) for a 301 base pair region of the G gene, indicating that the genetic diversity of IHNV within Alaska is notably lower than in the more southern portions of the IHNV North American range. Phylogenetic analysis of representative Alaskan sequences and sequences of 12 previously characterized IHNV strains from Washington, Oregon, Idaho, California (USA) and British Columbia (Canada) distinguished the isolates into clusters that correlated with geographic origin and indicated that the Alaskan and British Columbia isolates may have a common viral ancestral lineage. Comparisons of multiple isolates from the same site provided epidemiological insights into viral transmission patterns and indicated that viral evolution, viral introduction, and genetic stasis were the mechanisms involved with IHN virus population dynamics in Alaska. The examples of genetic stasis and the overall low sequence heterogeneity of the Alaskan isolates suggested that they are evolutionarily constrained. This study establishes a baseline of genetic fingerprint patterns and sequence groups representing the genetic diversity of Alaskan IHNV isolates. This information could be used to determine the source of an IHN outbreak and to facilitate decisions in fisheries management of Alaskan salmonid stocks. PMID- 10843555 TI - Multiplex PCR assay for ureC and 16S rRNA genes clearly discriminates between both subspecies of Photobacterium damselae. AB - A multiplex-PCR approach, employing 2 primer pairs directed to internal regions of the 16S rRNA and ureC genes, was utilized to analyze a collection of Photobacterium damselae strains, including 25 isolates of subspecies piscicida and 15 isolates of subspecies damselae. With this procedure, all the P. damselae subsp. damselae strains yielded 2 amplification products, one of 267 bp and the other of 448 bp, corresponding to internal fragments of the 16S rRNA and ureC genes, respectively. However, P. damselae subsp. piscicida isolates only showed the PCR product of 267 bp (16S rRNA fragment), indicating the absence of the urease gene in its genome. We have constructed a DNA probe directed to an internal region of the ureC gene, and corroborated by dot blot hybridization that the P. damselae subsp. piscicida lacks this gene, whereas it is present in the subspecies damselae. This constitutes the first successful discrimination between both subspecies using a PCR procedure, which could become a useful tool for diagnosis of pasteurellosis in the field. In addition, since these 2 subspecies have been shown to share nearly the same rrn operon sequence, our results provided evidence that one of the steps in the P. damselae speciation proccess included gain/loss events associated with the ure operon. PMID- 10843556 TI - Virulence properties of motile aeromonads isolated from farmed frogs Rana tigerina and R. rugulosa. AB - Virulence factors were compared in Aeromonas species isolated from clinically normal and septicaemic farmed frogs from Thailand. Haemolysin activities against frog erythrocytes were significantly different within the collection of aeromonads. Groups of high haemolytic activity (unspeciated Aeromonas, Au), moderate haemolytic activity (A. hydrophila), and low haemolytic activity (A. veronii biovar sobria, A. veronii biovar veronii, A. caviae, A. schubertii) were noted. DNA colony hybridisation studies revealed that Au isolates possessed a haemolysin gene (ASH1) which was not present in any of the other Thai aeromonads or type strains tested. Elastinolytic activity was demonstrated in 90% of the Au isolates, 60% of the A. hydrophila isolates and in none of the other motile aeromonads. The cytotoxic activity of the Aeromonas isolates varied according to the source of cells used in the assays. Cells from rainbow trout were extremely sensitive to Au toxins but less so to toxins produced by other species. In contrast mammalian cells showed very little sensitivity to Au toxins but were more sensitive to toxins produced by A. hydrophila. Selection of suitable assay substrates is therefore important. PMID- 10843557 TI - Ichthyophonus-like infection in wild amphibians from Quebec, Canada. AB - Myositis associated with infection by Ichthyophonus-like organisms was diagnosed in 35 of 260 (13%) wild amphibians collected in Quebec, Canada, from 1959 to 1964 (n = 30), and 1992 to 1999 (n = 230). Infection was diagnosed in 17 green frogs Rana clamitans, 9 wood frogs R. sylvatica, 4 red-spotted newts Notophthalmus viridescens, 3 bullfrogs R. catesbeiana, 1 spring peeper Pseudacris crucifer, and 1 pickerel frog R. palustris. The spring peeper and one of the bullfrogs were collected in 1964 from the Mont Saint-Hilaire Biosphere Reserve, indicating long term presence of the organism. Spores of the organisms invaded striated muscle fibers and were associated with variable degrees of granulomatous and eosinophilic inflammation. Infection was considered fatal in 2 green frogs, 1 wood frog, and 1 red-spotted newt. It was considered potentially significant in 3 additional green frogs in which up to 100% of the fibers of some muscles were replaced by spores associated with a severe granulomatous reaction. Ultrastructural features of Ichthyophonus-like spores included a thick trilaminated wall, a paramural cytoplasm, multiple nuclei, oval mitochondria with short tubulo-vesicular cristae and numerous ribosomes. This report represents 4 new host records and shows that ichthyophonosis is enzootic in amphibians from Quebec. PMID- 10843558 TI - Cytopathological observations on renal tubule epithelium cells in common carp Cyprinus carpio under Trypanoplasma borreli (Protozoa: Kinetoplastida) infection. AB - Cytological alterations in renal tubule epithelium cells of carp Cyprinus carpio infected with the blood flagellate Trypanoplasma borreli Laveran & Mesnil, 1901 were investigated during the course of a laboratory infection of a highly susceptible carp line. With the development of the parasitaemia, a hyperplasia of the interstitial renal tissue was induced, which resulted in a tubulus necrosis. Cytological changes were already seen in tubulus epithelium cells on Day 7 post injection (PI) of the parasite. The basilar invaginations of the cells fragmented and a swelling of mitochondria was noted. With increasing parasitaemia, on Days 14 and 21 PI, these changes progressed up to the loss of the basilar invagination and high amplitude swellings of mitochondria and deterioration of their internal membrane structures. Cells of the distal tubule segment reacted earlier and more rapidly than cells of the proximal tubule. The cytological alterations suggested a loss of function of the epithelum cells, which most likely resulted in impaired ionic and osmotic regulation of T. borreli-infected fishes. Our findings indicate that in response to the proliferation of the interstitial renal tissue cell structures of the renal tubule cells are altered quickly and in a progressive manner. PMID- 10843559 TI - Effect of salinity on hatching, survival and infectivity of Anguillicola crassus (Nematoda: Dracunculoidea) larvae. AB - The effect of salinity on hatching, larval survival and infectivity of Anguillicola crassus was studied under experimental conditions using eggs obtained from naturally infected eels. Egg hatching rate, second-stage larval survival and larval infectivity were maximal in fresh water and declined with increase in salinity. Larvae survived up to 100 d in fresh water, 70 d in 50 % sea water and 40 d in 100% sea water. Infectivity experiments demonstrated that salinity influenced transmission success throughout the life cycle by decreasing total infectivity of the larval population in utero within female A. crassus and when larvae were free-living in the aquatic environment. Infectivity was age dependent in relation to salinity. Larvae were infective to intermediate and paratenic hosts for up to 80 d in fresh water, 21 d in 50% sea water and up to 8 d in 100% sea water. The data confirm field observations that infection levels decrease with an increase in salinity. The study contributes to experimental verification of the colonization abilities of A. crassus and supports the hypothesis that A. crassus can be disseminated and transmitted in brackish water. The importance of regular monitoring and stringent hygiene practices in the transportation of eels is emphasized. PMID- 10843560 TI - Natural infection of the redclaw crayfish Cherax quadricarinatus with presumptive spawner-isolated mortality virus. AB - Crayfish farmers reported reduced tolerance of stress in specimens of Cherax quadricarinatus, which were formerly robust crayfish. Furthermore, one farmer reported a large reduction in yield with final harvest only equaling the stocking weight. Upon trapping, one-third of the crayfish regularly died overnight and a further one-third died on the sorting tray during sexing of juveniles (approximately 3 mo old). Histopathological examination revealed very light (1 or 2 cells per section) infections with Cherax giardiavirus and sometimes mild atrophy of hepatopancreatic cells. Gene probe analysis with a DIG-labeled spawner isolated mortality virus (SMV) probe demonstrated extensive positive signals in nuclei of many tissues. The hepatopancreas, the midgut, glands associated with the midgut, the epithelium of seminal ducts and follicle cells surrounding oocytes gave the strongest positive signals. Nuclei of the heart, haemocytes, connective tissue and subcutis gave positive signals in some individuals. Although signals were intense and extensive, cytolysis of infected cells was very limited. The possibility of cross infections of SMV between prawns and freshwater crayfish is of international quarantine significance. PMID- 10843561 TI - Comparison of infectious haematopoietic necrosis virus (IHNV) isolation on monolayers and in suspended cells. AB - A cell culture virus isolation procedure for infectious haematopoietic necrosis virus (IHNV) in the epithelioma papulosum cyprini cell line (EPC) is described. Ovarian fluid samples were collected from fish and tested for IHNV at 9 farms. The samples were inoculated in parallel on 24 h old EPC cell monolayers and in freshly trypsinized cells. The titre of the initial virus isolation and of first passages were compared using the 2 methods for each sample. Titres were consistently higher in suspended cells and this method also proved more sensitive for isolation of IHN virus from ovarian fluids of infected fish. PMID- 10843563 TI - On the generic placement of 'Livoneca sp.': a critique of Colorni et al. (1997). PMID- 10843562 TI - Treatment of Microcotyle sebastis (Monogenea: Polyopisthocotylea) infestation with praziquantel in an experimental cage simulating commercial rockfish Sebastes schlegeli culture conditions. AB - The antiparasitic efficacy of praziquantel against the blood-sucking polyopisthocotylean Microcotyle sebastis was tested in an experimental cage simulating commercial rockfish-culture conditions. Juvenile rockfish Sebastes schlegeli were separated into 4 pilot net-pens, and the fish in the groups were either fed a control diet (Group C), fed a praziquantel-adsorbed diet (Group F), bathed in 100 ppm praziquantel for 4 min (Group B), or bathed in 100 ppm praziquantel for 4 min and then fed a praziquantel-adsorbed diet (Group BF). The results of the present study indicate that feeding a praziquantel-adsorbed diet significantly reduces the abundance of M. sebastis infestation, and bathing in 100 ppm praziquantel for 4 min is effective for controlling M. sebastis infestation in practical rockfish culture systems. PMID- 10843564 TI - Longipinene derivatives from Santolina viscosa. AB - The neutral fraction of the hexane extract of the aerial parts of Santolina viscosa afforded eight new longipinene derivatives (1-8), oblongifolidiol (9), and several other known compounds. The structures of 1-8 were established by spectroscopic methods and chemical transformations. The vulgarone A-type structure previously reported for oblongifolidiol has been reassigned as the longipinene derivative 9 using 2D NMR, chemical correlations, and X-ray diffraction. PMID- 10843565 TI - Conversion of the iridoid glucoside antirrhinoside into 3-azabicyclo[3.3.0]octane building blocks. AB - The iridoid glucoside antirrhinoside (1) was transformed into polysubstituted 3 azabicyclo[3.3.0]octanes 3, 12, and 13 in 4 to 5 steps. Ozonolysis of the diacetonide of 1 and of its 7-deoxy-derivative 8 afforded cyclopentanoids 2 and 10, respectively. Conditions for the selective conversion of 2 and 10 into the corresponding ditosylates 4 and 11 were investigated. Cyclization of 4 and 11 was achieved with benzylamine and 2-methoxybenzylamine to yield bicyclic pyrrolidines 3, 12, and 13. Additional building blocks 14 and 15 were obtained by selective deprotection of the N-benzyl and isopropylidene moieties in 12 and 13, respectively. PMID- 10843566 TI - Indolyl carboxylic acids by condensation of indoles with alpha-keto acids. AB - The novel indole derivatives 2,2-bis(3,3'-indolyl)propionic acid (1); 1,1,1, tris(3,3',3"-indolyl)ethane (2); and 2,2-bis(3, 3'-indolyl)isocaproic acid (3) were isolated from solvent extracts of indole-supplemented supernatants of Escherichia coli and corynebacteria. The compounds were also obtained by chemical synthesis: compounds 1 and 2 from indole and pyruvic acid and compound 3 from indole and alpha-ketoisocaproic acid, following incubation at 37 degrees C in aqueous medium. Tryptophan and pyruvic acid gave the novel 2-(2 tryptophanyl)lactic acid (4). The condensation reaction between indoles and alpha keto acids was of general nature, and the mild reaction conditions suggested it may proceed in vivo. Examples for endogenous occurrence may be the neuro degenerative diseases phenylketonuria and maple syrup urine disease, both characterized by elevated blood levels of alpha-keto acids. PMID- 10843567 TI - New 7,20:14,20-diepoxy ent-kauranoids from Isodon xerophilus. AB - Three new 7,20:14,20-diepoxy-ent-kaurane diterpenoids, xerophilusins A-C (1-3), together with a known one, macrocalin B (4), were isolated from the leaves of Isodon xerophilus. Their structures were elucidated on the basis of their spectral properties and X-ray crystallographic analysis. Compounds 1, 2, and 4 showed significant cytotoxic activity against K562, HL-60, and MKN-28 cells. PMID- 10843568 TI - Cytonic acids A and B: novel tridepside inhibitors of hCMV protease from the endophytic fungus Cytonaema species. AB - Two novel human cytomegalovirus protease inhibitors, cytonic acids A (1) and B (2), have been isolated from the solid-state fermentation of the endophytic fungi Cytonaema sp. Their structures as p-tridepside isomers were elucidated by MS and NMR methods. PMID- 10843569 TI - Bioactive 12-oleanene triterpene and secotriterpene acids from Maytenus undata. AB - The aerial parts of Maytenus undata yielded four new 12-oleanene and 3,4-seco-12 oleanene triterpene acids, namely, 3-oxo-11alpha-methoxyolean-12-ene-30-oic acid (1), 3-oxo-11alpha-hydroxyolean-12-ene-30-oic acid (2), 3-oxo-olean-9(11), 12 diene-30-oic acid (3), and 3,4-seco-olean-4(23),12-diene-3, 29-dioic acid (20-epi koetjapic acid) (5), together with the known 3, 11-dioxoolean-12-ene-30-oic acid (3-oxo-18beta-glycyrrhetinic acid) (4), koetjapic acid (6), and the 12-oleanene artifact 3-oxo-11alpha-ethoxyolean-12-ene-30-oic acid (7). Koetjapic acid (6) inhibited the growth of Staphylococcus aureus, methicillin-resistant S. aureus, and Pseudomonas aeruginosa, with an MIC range of 3.125-6.25 microg/mL. The new 3,4-secotriterpene acid 20-epi-koetjapic acid (5) potently inhibited rat neonatal brain microglia phorbol ester-stimulated thromboxane B(2) (IC(50) = 0.5 microM) and superoxide anion (IC(50) = 1.9 microM) generation. PMID- 10843571 TI - Infraspecific variation of sulfur-containing bisamides from Aglaia leptantha. AB - Six new amides, leptaglin (1), hemileptaglin (2), aglanthin (3), agleptin (4), isoagleptin (5), and leptanthin (6), together with known lignans yangambin, eudesmin, grandisin (7), epigrandisin (8), and dehydrodiconiferyl alcohol, were isolated and identified from the lipophilic leaf and stem bark extracts of Aglaia leptantha. The dominating sulfur-containing bisamides contained either putrescine or the corresponding pyrrolidine ring as the diamine part, linked to phenylacetic and/or methylthiopropenoic acid moieties showing a remarkable infraspecific variation in eight individuals from two different habitats. Structures were determined by MS and NMR, including lanthanide-induced shifts. PMID- 10843570 TI - Isolation, structure determination, and biological activity of Lyngbyabellin A from the marine cyanobacterium lyngbya majuscula. AB - Lyngbyabellin A (1), a significantly cytotoxic compound with unusual structural features, was isolated from a Guamanian strain of the marine cyanobacterium Lyngbya majuscula. This novel peptolide is structurally related to dolabellin (2) in that both depsipeptides bear a dichlorinated beta-hydroxy acid and two functionalized thiazole carboxylic acid units. Its gross structure has been elucidated by spectral analysis, including 2D NMR techniques. The absolute stereochemistry of 1 was determined by chiral HPLC analysis of hydrolysis products and by characterization of the degradation products methyl 7,7-dichloro 3-hydroxy-2,2-dimethyloctanoate (3) and the corresponding acid 4. The total structure was further supported by molecular modeling studies. The isolation of 1 from L. majuscula once more supports the proposal that many compounds originally isolated from the sea hare Dolabella auricularia are of cyanobacterial origin. Lyngbyabellin A (1) was shown to be a potent disrupter of the cellular microfilament network. PMID- 10843572 TI - Production and structure elucidation of glycoglycerolipids from a marine sponge associated microbacterium species. AB - The bacterium Microbacterium sp., isolated from the sponge Halichondria panicea, produced four unusual cell-associated glycoglycerolipids and one diphosphatidylglycerol when grown on marine broth and on artificial seawater media. The lipids were isolated by chromatography on silica columns and their structures elucidated using a combination of multidimensional NMR and MS techniques. The main compound was 1-O-acyl-3-[alpha-glucopyranosyl-(1-3)-(6-O acyl-alpha-mannopyranosyl )]glycerol (GGL.2) with 14-methyl-hexadecanoic acid and 12-methyl-tetradecanoic acid positioned at C-6 of the mannose unit and at the glycerol moiety. Glycolipid production was correlated with growth and reached a maximum value of 200 mg/L when grown on artificial seawater medium with 20 g/L glucose. The main compound decreased the surface tension of water down to 33 mN/m and the interfacial tension of the water/n-hexadecane system down to 5 mN/m. In addition to this good surface-active behavior, the main glycoglycerolipid showed antitumor activities. PMID- 10843573 TI - Cytotoxic biflavonoids from Selaginella delicatula. AB - Four new biflavonoids-robustaflavone 4'-methyl ether (1), robustaflavone 7,4' dimethyl ether (2), 2",3" -dihydrorobustaflavone 7,4', -dimethyl ether (3), and 2",3" '-dihydrorobustaflavone 7,4', 7"-trimethyl ether (4)-as well as two known biflavonoids, robustaflavone and amentoflavone, and three caffeoylquinic acids, 3,5-di-O-caffeoylquinic acid, 3, 4-di-O-caffeoylquinic acid, and 4,5-di-O caffeoylquinic acid, were isolated from Selaginella delicatula. The structures of the new compounds were established by spectroscopic analysis and chemical modification. The cytotoxic activity of these compounds on various tumor cell lines was evaluated, and both compounds 1 and 3 significantly suppressed the growth of Raji and Calu-1 tumor cell lines. PMID- 10843574 TI - Aminosterols from the dogfish shark Squalus acanthias. AB - Seven new aminosterols related to squalamine (8) were isolated from the liver of the dogfish shark Squalus acanthias. Their structures (1-7) were determined using spectroscopic methods, including 2D NMR and HRFABMS. These aminosterols possess a relatively invariant cholestane skeleton with a trans AB ring junction, a spermidine or spermine attached equatorially at C3, and a steroidal side-chain that may be sulfated. The structure of the lone spermine conjugate, 7 (MSI-1436), was confirmed by its synthesis from (5alpha,7alpha, 24R)-7-hydroxy-3 ketocholestan-24-yl sulfate. Some members of this family of aminosterols exhibit a broad spectrum of antimicrobial activity comparable to squalamine. PMID- 10843575 TI - Phytochemical investigation of Aglaia rubiginosa. AB - The phytochemical investigation of a methanolic leaf extract of Aglaia rubiginosa furnished 15 isoprenoid constituents, eight of which represented new natural entities. Two androstane derivatives (1 and 2), previously synthesized, and also obtained by microbiological transformations; an extraordinary 17-octanor cycloartane-ring-A-seco acid (3); four cycloartane-type triterpenes (4-7); and three unusual cholesterol derivatives (8-10) were isolated, along with two known dammaranes (11 and 12), a stigmastandiol (13), and beta-sitosterol and its beta-D glucoside. Spectroscopic structure elucidation of the new natural products (1-3, 6, 7, 8-10) is described. PMID- 10843576 TI - Studies directed toward the synthesis of cryptoheptine. AB - Synthesis of 5,10-dihydro-10-methylindolo[3, 2-b][1]benzazepin-12(11H)-one (2), an isomer of the reported structure for cryptoheptine (1), is presented. Attempts to convert 2 to 1 led to 10-methylindolo[3,2-b][1]benzazepin-12-one (10), an oxidation product of 2 and presumably 1. These results highlight the potential instability of cryptoheptine and cast doubt on its proposed structure. PMID- 10843577 TI - Triterpene saponins in the defensive secretion of a chrysomelid beetle, Platyphora ligata. AB - The secretion of the defensive glands of adults of the chrysomelid beetle Platyphora ligata from Panama has been shown to contain, besides chlorogenic acid (1) and a mixture of phosphatidylcholines, two new oleanane triterpene saponins, named ligatosides A and B. Their structures were established as 3-O-beta-D glucuronopyranosyl-16alpha,23-dihydroxyoleanol ic acid-28-O-2-(3,4 dimethoxybenzoyl)-beta-D-glucopyranoside (2) and 3-O-beta-D-glucopyranosyl-(1- >2)-beta-D-glucuronopyranosyl-16a lpha, 23-dihydroxyoleanolic acid-28-O-2-(3, 4 dimethoxybenzoyl)-beta-D-glucopyranoside (3), respectively, by a combination of extensive 1D and 2D NMR methods (COSY, HMQC, HMBC, and TOCSY) and FABMS. This is the first report of triterpene saponins in the defensive secretion of an insect. PMID- 10843578 TI - Podocarpane-type trinorditerpenes from the bark of Taiwania cryptomerioides. AB - Five new podocarpane-type trinorditerpenes were isolated from the bark of Taiwania cryptomerioides. Structures of 1beta, 14-dihydroxy-13-methoxy-6,8,11,13 podocarpatetraene (1), 1beta, 14-dihydroxy-13-methoxy-8,11,13-podocarpatriene (2), 13-hydroxy-12-methoxy-8,11,13-podocarpatrien-3-one (3), 14-hydroxy-13 methoxy-1,8,11,13-podocarpatetraen-3-one (4), and 13-methoxy-8,12-podocarpatriene 11,14-dione (5) were determined by NMR, MS, and other spectral and chemical evidence. PMID- 10843580 TI - Novel cytotoxic diterpenes from Casearia arborea. AB - Cytotoxicity-guided fractionation of the dichloromethane-methanol extract of the roots of Casearia arborea yielded five novel clerodane diterpenes, casearborins A E (1-5), as well as cucurbitacin B. The presence of cucurbitacins glycosides was also detected. The absolute configuration of casearborin E was determined by X ray crystallography. PMID- 10843579 TI - Constituents of Eremurus chinensis. AB - A novel bianthraquinone glycoside, 8-O-beta-D-glucopyranosyl-1,1', 8'-trihydroxy 3,3'-dimethyl-2,7'-bianthraquinone (1); two naphthalene derivatives, 2-acetyl-1 hydroxy-8-methoxy-3-methylnaphthalene (2) and 2-acetyl-1, 8-dimethoxy-3 methylnaphthalene (3); and a novel pre-anthraquinone, 1-oxo-4(S),9-dihydroxy-8 methoxy-6-hydroxymethyl-1,2,3, 4-tetrahydroanthracene (4), were isolated from Eremurus chinensis. Their structures were established by spectroscopic and chemical methods. In addition, the known compounds chrysophanol, chrysophanol 8 methyl ether, aloesaponol III 8-methyl ether (5), and 10-(chrysophanol-7'-yl)-10 hydroxychrysophanol-9-anthrone were also isolated and identified from this plant. PMID- 10843581 TI - A new flavonoid glycoside from Centaurea horrida. AB - A new natural compound, horridin (1), was isolated from the aerial parts of Centaurea horrida. Its structure as quercetin 3-O-alpha-L-rhamnopyranosyl-(1-->2) alpha-L-rhamnopyranoside was determined by spectroscopic methods, including 2D NMR. PMID- 10843582 TI - N-acetyl-gamma-hydroxyvaline lactone, an unusual amino acid derivative from a marine streptomycete. AB - We report the isolation and structure elucidation of N-acetyl-gamma-hydroxyvaline lactone (1) from a streptomycete obtained from marine sediments collected in the north coastline of Sao Paulo State, Brazil. PMID- 10843583 TI - Identification and total synthesis of a novel dimethylated fatty acid from the Caribbean sponge Calyx podatypa. AB - The dimethylated fatty acid 9,13-dimethyltetradecanoic acid was identified for the first time in nature in the Caribbean sponge Calyx podatypa where it occurs together with the rare 10, 13-dimethyltetradecanoic acid. The characterization of the novel compound was accomplished using GC-MS, pyrrolidide derivatization, and a five-step total synthesis starting with 8-bromooctanoic acid. The first racemic total synthesis for the rare 10, 13-dimethyltetradecanoic acid is also described. PMID- 10843584 TI - Polyhydroxylated sterols from the octocoral Dendronephthya gigantea. AB - Two new polyhydroxylated sterols, dendronesterols A (1) and B (2), have been isolated from the octocoral Dendronephthya gigantea. The structures of 1 and 2 were proposed on the basis of extensive NMR experiments. Compound 2 was found to be weakly cytotoxic toward L1210 cells. PMID- 10843585 TI - Rotundines A-C, three novel sesquiterpene alkaloids from Cyperus rotundus. AB - Rotundines A (1), B (2), and C (3), three novel sesquiterpene alkaloids with an unprecedented carbon skeleton, were isolated from the rhizomes of Cyperus rotundus. The structures of 1-3 were elucidated by spectral and chemical methods. PMID- 10843587 TI - Cytotoxic lavandulyl flavanones from Sophora flavescens. AB - Two new lavandulylated flavanones, (2S)-2'-methoxykurarinone (1) and (-) kurarinone (2), were isolated from the root of Sophora flavescens, together with two known lavandulyl flavanones, sophoraflavanone G (3) and leachianone A (4), and two known isoflavonoids, formononetin and l-maakiain. The structures of 1 and 2 were determined on the basis of optical rotation and spectral evidence and by comparison with known compounds. Compounds 1-4 exhibited cytotoxic activity against human myeloid leukemia HL-60 cells. PMID- 10843586 TI - Thonningianins A and B, new antioxidants from the African medicinal herb Thonningia sanguinea. AB - Two new ellagitannins, thonningianins A (1) and B (2), have been isolated from the African medicinal herb Thonningia sanguinea and their structures elucidated by interpretation of spectroscopic data. Both 1 and 2 showed strong free radical scavenging activity against 1,1-diphenyl-2-picrylhydrazyl (DPPH) as shown by ESR analysis. PMID- 10843588 TI - Hachijodines A-G: seven new cytotoxic 3-alkylpyridine alkaloids from two marine sponges of the genera Xestospongia and amphimedon. AB - Seven cytotoxic 3-alkylpyridine alkaloids, hachijodines A-G, have been isolated from two marine sponges of the genera Xestospongia and Amphimedon. Their structures were determined on the basis of spectral data. These alkaloids are moderately cytotoxic against P388 murine leukemia cells with IC(50) values of 1.0 2.3 microg/mL. PMID- 10843589 TI - Characterization of the major metabolite of sampangine in rats. AB - Sampangine (1) is a plant-derived antifungal copyrine alkaloid extracted from the stem bark of Cananga odorata. Although it possesses potent in vitro antifungal activity, 1 is devoid of significant and reproducible in vivo activity in a mouse model of cryptococcosis. Speculating that the lack of in vivo activity could be due to metabolism, a study was undertaken to begin to develop an understanding of the pharmacokinetics, and particularly metabolism of 1. Following intraperitoneal administration of 1 to rats, urine was collected, extracted, and chromatographed over a reversed-phase C(18) silica column to yield the major metabolite, SAM MM1 (2), which was identified by NMR and MS to be an O-glucuronide conjugate of sampangine. In addition, two other unstable, structurally uncharacterized minor metabolites were produced, as evidenced by HPLC analysis. Evaluation of the antifungal and antibacterial activities of 2 showed it to have remarkable in vitro activity against Cryptococcus neoformans. PMID- 10843590 TI - 10-Hydroxydarlingine, a new tropane alkaloid from the Australian proteaceous plant triunia erythrocarpa. AB - Triunia erythrocarpa was identified as containing alkaloids during chemical screening of Queensland Proteaceae using Dragendorff's reagent. A new tropane, 10 hydroxydarlingine (1), and the known tropane, darlingine (2), were isolated from the leaves of T. erythrocarpa. The absolute stereochemistry of 10 hydroxydarlingine (1) was assigned using the advanced Mosher method. T. erythrocarpa is only the seventh member of the Proteaceae to have been shown to produce alkaloids. PMID- 10843591 TI - An antimicrobial C(14) acetylenic acid from a marine sponge Oceanapia species. AB - A C(14) acetylenic acid has been isolated as an antimicrobial principle from a marine sponge Oceanapia sp. Its structure was determined on the basis of spectral data. PMID- 10843592 TI - Constituents from the leaves of Aristolochia elegans. AB - One new biphenyl ether, aristogin C (1), and two new porphyrins, aristophylls A (2) and B (3), as well as 11 known compounds, were isolated from the leaves of Aristolochia elegans. Their structures were elucidated according to the spectroscopic (NMR and MS) analyses or by comparison with literature values. PMID- 10843593 TI - Polyoxygenated dysidea sterols that inhibit the binding of [I125] IL-8 to the human recombinant IL-8 receptor type A. AB - The combined CH(2)Cl(2) and MeOH crude extract of a new species of the marine sponge Dysidea, collected in Northern Australia was found to inhibit the binding of [I125] interleukin-8 [IL-8] to the human recombinant IL-8 receptor type A at 500 microg/mL. Bioassay-guided fractionation led to the isolation of three new polyoxygenated sterols 3, 4, and 5. Their structures were assigned on the basis of 1D and 2D NMR experiments, and relative stereochemistries were established by ROESY correlations and analysis of coupling constants. The IC(50) values for inhibition of IL-8Ra for sterols 3, 4, and 5 were 20, 5.5, and 4.5 microM, respectively. PMID- 10843594 TI - New diketopiperazine alkaloids from Penicillium fellutanum. AB - Four new diketopiperazine alkaloids (1-4) were isolated from cultures of Penicillium fellutanum, and their structures were determined by MS and NMR measurements. PMID- 10843595 TI - A new triterpenoid saponin from Isolatocereus dumortieri. AB - A new triterpenoid saponin, named dumortierinoside A, was isolated from Isolatocereus dumortieri. The structure was determined as dumortierigenin 3-O alpha-L-rhamnopyranosyl (1-->2)-beta-D-glucopyranosyl(1-->2)-beta-D glucuronopyranoside (1) on the basis of NMR and mass spectroscopy. PMID- 10843596 TI - Mycalamides C and D, cytotoxic compounds from the marine sponge Stylinos n. species. AB - The new cytotoxic compounds, mycalamides C (3) and D (4), have been isolated from the marine sponge Stylinos n. sp., along with the known theopederin E (1) and mycalamide A (2). PMID- 10843597 TI - Mycalamide D, a new cytotoxic amide from the New Zealand marine sponge Mycale species. AB - A new mycalamide, mycalamide D (3), has been isolated from the New Zealand marine sponge Mycale sp. This new metabolite, in which the C13-O-methyl group of mycalamide A (1) is replaced by a hydrogen atom, was found to be cytotoxic to a range of mammalian cell lines, with a potency approximately 20-fold less than that of 1. PMID- 10843598 TI - Three new flavonol galloylglycosides from leaves of Acacia confusa. AB - Myricetin 3-O-(2"-O-galloyl)-alpha-rhamnopyranoside 7-methyl ether (1), myricetin 3-O-(3"-galloyl)-alpha-rhamnopyranoside 7-methyl ether (2), and myricetin 3-O (2",3" -di-O-galloyl)-alpha-rhamnopyranoside (3), three new flavonol galloylglycosides, were isolated from leaves of Acacia confusa sampled from Chaoushi in the north of Taiwan. Their structures were established by analysis of spectroscopic data, and the compounds were evaluated for anti-hatch activity against brine shrimp. PMID- 10843599 TI - Evidence for an NIH shift as the origin of the apparently anomalous distribution of deuterium in estragole from Artemisia dracunculus. AB - Feeding experiments of [2-(2)H]- and [4'-(2)H]phenylalanine in Artemisia dracunculus validate the hypothesis that the marked difference in deuterium content at the natural abundance level between the aromatic carbons of estragole (1) is due to an NIH shift during the hydroxylation of the benzene ring. PMID- 10843600 TI - Petersaponins III and IV, triterpenoid saponins from Petersianthus macrocarpus. AB - Two new triterpenoid saponins, petersaponins III and IV (1 and 2), were isolated from an n-butanol extract of the bark of Petersianthus macrocarpus. They possess 21-O-benzoyl-22-O-acetylbarringtogenol C and 21-O-2-furoxyl-22-O tigloylbarringtogenol C as the aglycon, respectively. For both 1 and 2, the trisaccharide moiety linked to C-3 of the aglycon consists of D-glucuronic acid, D-xylose, and D-galactose, while a L-rhamnose unit is linked to C-28. The structures of 1 and 2 were elucidated by extensive NMR experiments including (1)H (1)H (COSY, HOHAHA, NOESY) and (1)H-(13)C (HMQC and HMBC) spectroscopy and by chemical evidence. PMID- 10843601 TI - New taxanes with an opened oxetane ring from the roots of Taxus mairei. AB - Two novel taxoids, taxumairols N (1) and O (2), have been isolated from extracts of the roots of Taxus mairei. The structures of 1 and 2 were identified as 7beta,9alpha,10beta,13alpha-tetraacetoxy-2alp ha, 4alpha,5alpha,20 tetrahydroxytax-11-ene and 7beta,9alpha,10beta, 13alpha-tetraacetoxy 1beta,2alpha,4alpha,5alp ha, 20-pentahydroxytax-11-ene on the basis of 1D and 2D NMR techniques including COSY, HMQC, HMBC, and NOESY experiments. PMID- 10843602 TI - Two Alangium alkaloids from Alangium lamarckii. AB - Two new Alangium alkaloids, 1',2'-dehydrotubulosine (1) and alangine (2), were isolated from the dried fruits of Alangium lamarckii along with tubulosine (3), isotubulosine (4), deoxytubulosine, cephaeline, isocephaeline, psychotrine, neocephaeline, 10-O-demethylcephaeline, 2'-N-(1"-deoxy-1" -beta-D fructopyranosyl)cephaeline, protoemetine, protoemetinol, salsoline, and alangiside. The structures of the new alkaloids (1 and 2) were determined by spectroscopic and chemical means. PMID- 10843604 TI - Computational explorations of vinylcyclopropane-cyclopentene rearrangements and competing diradical stereoisomerizations AB - The rearrangements and stereoisomerizations of four systems, vinylcyclopropane, 4 tert-butylvinylcyclopropane, 5-methylvinylcyclopropane, and 2,5 dimethylvinylcyclopropane, as well as a variety of deuterated derivatives and 1- and 2-methyl-, methoxy-, difluoro-, and amino-substituted species, were studied by density functional theory calculations using the B3LYP functional and the 6 31G basis set. Energies were evaluated with CASSCF(4, 4)/6-31G single point calculations. The major product is obtained by the si pathway. Structures on this path are essentially pure diradical in character. Higher energy diradical species and intermediates are responsible for the scrambling of the stereochemistry. The stereoselectivity of the reaction is increased by substituents which increase the relative energy of the species involved in competing stereoselectivities. The computed secondary kinetic isotope effects reproduce the experimental values reported in the literature. PMID- 10843603 TI - Recent advances in the chemistry of taxol. AB - The history of the development of the important anticancer drug taxol (1) is briefly described, and recent studies of its chemistry and tubulin-binding conformation are then presented. Topics discussed include side chain attachment to baccatin III (3a), the effect of oxygenation of the taxane ring system on bioactivity, the importance of the oxetane ring for bioactivity, the synthesis of a C-6/C-4 bridged analogue, and the conformation of the side chain when taxol is bound in a complex with polymerized tubulin. PMID- 10843605 TI - Electronic interactions in a new fullerene dimer: C(122)H(4), with two methylene bridges AB - The isolation of a new fullerene dimer, C(122)H(4), and its structural characterization by (13)C NMR and (1)H NMR spectroscopy and by UV/vis and IR spectroscopy are reported. The structure of this dimer consists of two fullerene cages, which are directly connected through two C-C bonds and two methylene bridges. Consequently, adjacent hexagonal faces of the two fullerene cages are arranged in a face to face manner. Molecular orbital calculations indicate that the proximity of the fullerene cages results in significant through space overlap in both the HOMO and LUMO. As a consequence of this overlap, the electrochemistry of the dimer shows electronic communication with stepwise reduction of each cage. PMID- 10843606 TI - Highly regioselective synthesis of 3,4-disubstituted 1H-pyrrole. AB - A highly regioselective method for the synthesis of 3, 4-disubstituted 1H pyrroles has been developed employing the ipso-directing property of a trimethylsilyl group. As a key starting material in this study, the known 3,4 bis(trimethylsilyl)-1H-pyrrole (3), was protected with carefully chosen groups, namely tert-butoxycarbonyl, N,N-dimethylaminosulfonyl, p-toluenesulfonyl, and triisopropylsilyl. A highly regioselective monoiodination of these 1-protected pyrroles was achieved by reaction with iodine-silver trifluoroacetate at low temperatures. Subsequent palladium-catalyzed cross-coupling reactions afforded 1 protected-4-substituted 3-trimethylsilyl-1H-pyrroles, which again underwent further room-temperature ipso-iodination and palladium-catalyzed cross-coupling reactions to provide symmetrical and unsymmetrical 1-protected-3,4-disubstituted 1H-pyrroles. Deprotection of 1-(tert-butoxycarbonyl) and 1-(N, N dimethylaminosulfonyl) groups was found to be nontrivial. The 1-(p toluenesulfonyl) protecting group was eventually proved to be superior to other protection groups, because it was readily removed after stepwise ipso monoiodinations and palladium-catalyzed cross-coupling reactions. PMID- 10843607 TI - C(2)-Symmetric bis-sulfoxide: A novel chiral auxiliary for asymmetric desymmetrization of cyclic meso-1,2-diols AB - A new heterocyclic compound, C(2)-symmetric bis-sulfoxide 1, has been found to be an efficient chiral auxiliary for asymmetric desymmetrization of cyclic meso-1,2 diols via diastereoselective acetal fission. Both (R,R)- and (S,S)-1 are readily synthesized with high optical purity via asymmetric oxidation of 1, 5 benzodithiepan-3-one (2). After acetalization of meso-1,2-diols 6a-e and a mono TMS ether 6f with this chiral auxiliary 1, the resulting acetals 7a-f were subjected to base-promoted acetal fission upon treatment with potassium hexamethyldisilazide (KHMDS) followed by acetylation or benzylation to give the desymmetrized diol derivatives 8a-f with high diastereoselectivity. The chiral auxiliary 1 is readily removed by acid-promoted hydrolysis and can be recovered without a loss in enantiomeric excess. PMID- 10843608 TI - Dithia-crown-annelated tetrathiafulvalene disulfides: synthesis, electrochemistry, self-assembled films, and metal ion recognition AB - The synthesis and electrochemistry of a series of tetrathiafulvalene (TTF) and dithia-crown-TTF derivatives attached with one or two disulfide group(s) 7a-f are reported. The self-assembled monolayers (SAMs) of these TTF disulfides on gold were prepared and characterized by reflection-absorption infrared spectroscopy. The SAMs are extremely stable under a wide variety of conditions and over extended periods of time and show remarkable electrochemical stability upon repeated potential scans. SAMs of the crown-TTF disulfides 7c,d,f can recognize alkali metal ions, and the process can be monitored following the electrochemical potential shift of the surface-confined TTF group. PMID- 10843609 TI - Synthesis of Calix AB - The formation of p-tert-butylcalix[4]arene mono-, bis-, tris-, and tetrakistriflates 3, 2, 4, and 5, respectively, and their respective reactions, under typical Pd-catalyzed carbonylative, Suzuki-Miyaura coupling, or deoxygenation conditions are described. A novel, nonsolvent-derived 1:1 clathrate (6) of benzophenone and 3 was formed from the palladium-catalyzed carbonylative reaction of phenylboronic acid and 2. The X-ray crystal structure of this first nonsolvent-derived clathrate of a calix[4]arene derivative is reported. Another 1:1 clathrate of triethylamine and 3 was formed during the attempted Pd-catalyzed deoxygenation of 2. PMID- 10843610 TI - Absolute proton affinity of some polyguanides AB - The problem of the absolute proton affinity (APA) of some polyguanides is addressed by the MP2(fc)/6-311+G//HF/6-31G theoretical model. It is shown that the linear chain polyguanides exhibit increased basicity as a function of the number of guanide subunits. However, the saturation effect yields an asymptotic APA value of 254 kcal/mol. Branched polyguanides on the other hand have higher APAs than their linear counterparts. The largest proton affinity is found in a doubly bifurcated heptaguanide, being as high as 285 kcal/mol, thus potentially representing one of the strongest organic bases. Finally, it is found that all polyguanides protonate at imino nitrogen atoms, since they are apparently susceptible the most to the proton attack. The origin of their very high intrinsic basicity is traced down to a dramatic increase in the resonance interaction of the corresponding conjugate bases. For instance, the increase in the resonance energy in the protonated guanidine is estimated to be in a range of 24-27 kcal/mol, which is higher than the aromatic stabilization in benzene. The proton affinity of some polycyclic guanides including Schwesinger proton sponge and porphine is briefly discussed. PMID- 10843611 TI - Stereoselective allylation of 4-oxoazetidine-2-carbaldehydes. Application To the stereocontrolled synthesis of fused tricyclic beta-lactams via intramolecular diels-alder reaction of 2-azetidinone-tethered trienes AB - Allylation reactions of racemic and optically pure 4-oxoazetidine-2-carbaldehydes were investigated both under anhydrous conditions and in aqueous media. Different Lewis acid or metal mediators showed varied diastereoselectivities on product formation during allylation reactions of the above aldehydes with allyltrimethylsilane, allyltributylstannane, or allyl bromide. Under standard reaction conditions, tin(IV) chloride-promoted addition of allyltrimethylsilane to 4-oxoazetidine-2-carbaldehydes provided the highest diastereoselectivity and the best yield. Boron trifluoride-diethyl ether-promoted reaction of allyltributylstannane provided slightly lower diastereoselectivity with the same facial preference. Indium-promoted allylation showed a reverse diastereofacial preference, although the observed selectivity is not synthetically useful. The mesylates of these homoallylic alcohols were used for the stereoselective preparation of cis-4-butadienyl-2-azetidinones. Interestingly, mesylates having an extra alkene or alkyne tether at position 1 or 3 of the beta-lactam ring, on heating in a sealed tube with equimolecular amounts of DBU in toluene, yielded fused tricyclic 2-azetidinones through a tandem one-pot elimination intramolecular Diels-Alder reaction. PMID- 10843612 TI - Kinetics and mechanism of the oxidation of substituted benzaldehydes by benzyltrimethylammonium chlorobromate AB - The oxidation of 35 monosubstituted benzaldehydes by benzyltrimethylammonium chlorobromate (BTMACB) in aqueous acetic acid leads to the formation of the corresponding benzoic acids. The reaction is first order with respect to both benzaldehyde and BTMACB. The reaction failed to induce the polymerization of acrylonitrile. There is no effect of benzyltrimethylammonium chloride or potassium bromide on the reaction rate. The oxidation of [(2)H]benzaldehyde (PhCDO) indicated the presence of a substantial kinetic isotope effect. The effect of solvent composition indicated that the reaction rate increases with an increase in the polarity of the solvent. The rates of oxidation of meta- and para substituted benzaldehydes were correlated in terms of Charton's triparametric LDR equation, whereas the oxidation of ortho-substituted benzaldehydes was correlated with the tetraparametric LDRS equation. The oxidation of para-substituted benzaldehydes is more susceptible to the delocalization effect, whereas the oxidation of ortho-and meta-substituted compounds displayed a greater dependence on the field effect. The positive value of eta suggests the presence of an electron-deficient reaction center in the rate-determining step. The reaction is subjected to steric hindrance by the ortho substituents. PMID- 10843613 TI - Novel chiral bisoxazoline ligands with a biphenyl backbone: preparation, complexation, and application in asymmetric catalytic reactions AB - Novel C(2)-symmetric chiral bisoxazoline ligands 1 were easily prepared from enantiomerically pure 2-amino alcohols and achiral 2, 2'-biphenyldicarboxylic acid via the corresponding amide and mesylate as intermediates. Since these ligands bear only two ortho-substituents on the biphenyl backbone, the biphenyl axis is not fixed, and the two diastereomers of these ligands exist in equilibrium in solution. Interestingly, when the ligands 1 were coordinated with a metal ion, only one of the two possible diastereomer complexes, an (S,aS,S) complex, can be formed depending on the combination of the ligand and the metal ion. Thus, copper(I) afforded only the (S,aS,S)-complexes with all ligands 1, while zinc(II), palladium(II), and silver(I) afforded the (S,aS, S)-complexes as the sole product only with 1b, which has a bulky tert-butyl group on the oxazoline ring, and a mixture of the two diastereomer complexes with 1a,c,d. The copper(I)-catalyzed asymmetric cyclopropanation of styrene with diazoacetate proceeded successfully with these ligands and good to excellent enantioselectivities were afforded. PMID- 10843614 TI - Mechanism of selective oxidation of organic sulfides with Oxo(salen)chromium(V) complexes AB - The selective oxidation of organic sulfides to sulfoxides by oxo(salen)chromium(V) complexes in acetonitrile is overall second-order, first order each in the oxidant and the substrate. The rate constant, k(2), values of several para-substituted phenyl methyl sulfides correlate linearly with Hammett sigma constants and the rho values are in the range of -1.3 to -2.7 with different substituted oxo(salen)chromium(V) complexes. The reactivity of different alkyl sulfides is in accordance with Taft's steric substituent constant, E(S). A mechanism involving direct oxygen atom transfer from the oxidant to the substrate rather than electron transfer is envisaged. Correlation analyses show the presence of an inverse relationship between reactivity and selectivity in the reaction of various sulfides with a given oxo(salen)chromium(V) complex and vice versa. Mathematical treatment of the results shows that this redox system falls under strong reactivity-selectivity principle (RSP). PMID- 10843615 TI - One-Pot synthesis of N-heterocyclic compounds from cyclopropenethione derivatives AB - The one-pot reaction of 2-tert-butylthio-3-phenylcyclopropenethione (1a) and its 3-(2-thienyl) derivative (1b) with lithium pyrrolidinide at -70 degrees C, followed by methylation with methyl iodide, gives 6-methylthio-5-phenyl-2,3 dihydro-1H-pyrrolizine (2a) and its 5-(2-thienyl) derivative (2b), respectively. The reaction of 2-tert-butylthio-3-(pyrrolidin-1-yl)cyclopropenethione (1c) with phenyllithium gives also 2a in a high yield under similar conditions, and the reactions of 1a with N-lithium salts of 3-pyrroline, hexamethyleneimine, indoline, and carbazole, piperidine-potassium tert-butoxide mixture, and phenyllithium give 6-methylthio-5-phenyl-3H-pyrrolizine (3), 2-methylthio-3 phenyl-6,7, 8,9-tetrahydro-5H-pyrrolo[1,2-a]azepine (5), 6-tert-butylthio-5 methylthio-4-phenyl-1,2-dihydro-6H-pyrrolo[3,2, 1-ij]quinoline (6), 4-tert butylthio-5-methylthio-6-phenyl-4H-pyrido[3,2,1-jk]carbazole (7), 2-methylthio-3 phenyl-5,6,7,8-tetrahydroindolizine (4), and 1-tert-butylthio-2-methylthio-3 phenylindene (9), respectively. The structures of 2a and 3 were determined by X ray analyses of their tricarbonylchromium complexes. PMID- 10843616 TI - Synthesis of a C(29)-C(51) subunit of spongistatin 1 (Altohyrtin A) starting from (R)-3-benzyloxy-2-methylpropan-1-ol. AB - A protected C(29)-C(51) subunit ((+)-38) of spongistatin 1 has been obtained. Key steps involve the aldol condensation of (3S, 4R)-3-methyl-7-[(p methoxybenzyl)oxy]-4-[(triethylsilyl)oxy]octan- 2-o ne ((-)-6) with (tert butyl)dimethylsilyl 4-deoxy-2, 3-di-O-(methoxymethyl)-4-methyl-6-O-(tert butyl)dimethylsilyl)-bet a-D -glycero-L-gluco-heptodialdo-1,5-pyranoside ((+)-7) and a C-glycosidation of (4R,7R&S,E)-7, 8-dichloro-2-methylidene-1 (trimethylsilyl)oct-5-en-4-yl p-methoxybenzoate (16). Aldehyde (+)-7 was derived from (R)-3-benzyloxy-2-methylpropan-1-ol ((+)-10) in 13 formal steps but requiring the isolation of five intermediate products only. The longest linear synthetic scheme converts (+)-10 into (+)-38 in 2% overall yield (isolation of 11 intermediate products). PMID- 10843617 TI - Synthesis and supramolecular characterization of a novel class of glycopyranosyl containing amphiphiles. AB - A novel class of glycopeptidolipids is described, which potentially can be used as a novel antigen-delivery system. The compounds have been prepared by a combination of solid-supported and solution-based methods. The use of the orthogonally protected FmocLysDde derivative provided an opportunity to incorporate two different types lipids. It was found that the model compound 1 forms aggregates in aqueous media which can be described as rod or tubelike structures. The aggregates can be stabilized by topotactic photopolymerization. Studies on the structural analogues 2-5 revealed the effect of the carbohydrate, peptide, and lipid moiety on the aggregation properties. It is concluded that none of the structure elements can lay claim to be exclusively important for the formation of highly organized aggregates such as tubes, fibers, or helical ribbons from 1, but the presence of all of these structural elements afforded the most uniformly shaped extended structures. PMID- 10843618 TI - X-ray structures and anionotropic rearrangements of Di-tert-butyl-substituted thiiranium and thiirenium ions. A structure-reactivity relationship AB - The X-ray structures of c-2,t-3-di-tert-butyl-r-1-methylthiiranium 8 BF(4)(-), t 2,t-3-di-tert-butyl-r-1-methylthiiranium ion 10 BF(4)(-), and 2,3-di-tert-butyl-1 methylthiirenium 11 BF(4)(-) have been determined. The DeltaG()(298) values for the rearrangements from the cis and the trans tert-butyl groups of 8 SbCl(6)(-) to thietanium ion (two intramolecular S(N)2 displacements) and for the rearrangement of 11 SbCl(6)(-) to thietium ion (an intramolecular S(N)2-Vin displacement) are linearly correlated with the strengths of the C-S breaking bonds, suggesting that the two mechanisms are, in the absence of steric hindrance, uniquely governed by the nucleofugality of the sulfonium leaving group. PMID- 10843619 TI - Regio- and endo-selective AB - Irradiation of various meta- and para-substituted homobenzoquinones with ethyl vinyl ether gave the [2 + 2] photoadducts, tricyclic diones, regio- and endo selectively and in good yields. The tricyclic skeleton has an anti-form built by the addition of ethyl vinyl ether from the less hindered side of homoquinones. All of the CH(3), Cl, Br, and CH(3)O substituents at the reacting C=C double bond afforded head-to-head (HH) addition predominantly. In the case of CH(3), Cl, and Br, the ethoxy group was oriented in the endo-position, while the CH(3)O substituent led to a 1/5 mixture with the exo-isomer. It was also found that the Br-substituted [2 + 2] adducts undergo a facile skeletal rearrangement, being converted into dihydro-o-benzoquinone monomethide derivatives for para substitution and dihydrobenzofuran derivatives for meta-substitution, probably under the influence of the in situ generated HBr. Intramolecular [2 + 2] photocycloaddition of an alkenylhomobenzoquinone afforded a tetracyclic dione. PMID- 10843620 TI - Oxy-cope rearrangements of bicyclo[3.2.0]heptenones. Synthesis of bicyclo[4.2.1]non-1(4)-en-6-ones and bicyclo[5.2. 1]dec-1(10)-en-5-ones. AB - 6-exo-Methylbicyclo[3.2.0]hepten-7-ones and their 2-alkylidene analogues are readily prepared from dialkyl squarates. These compounds undergo facial oxy-Cope ring expansions upon treatment with vinyllithium; the former leads to bicyclo[4.2. 1]non-1(4)-en-6-ones and the latter to the first examples of bicyclo[5.2.1]dec-1(10)-en-5-ones, compounds having exceptionally strained bridgehead double bonds. The transformations are controlled by the 6-exo-methyl group in the starting material along with the substituent at position-1 (bridgehead) which force attack of the lithium reagent from the concave face of the starting material, thus allowing the cyclopentenyl or alkylidene groups to participate in the sigmatropic event. PMID- 10843621 TI - Open analogues of arcyriaflavin A. Synthesis through Diels-Alder reaction between maleimides and 1-aryl-3-tert-butyldimethylsiloxy-1, 3-butadienes. AB - The preparation of a range of open analogues of arcyriaflavin A is described. The synthetic approach is based on the use of perhydroisoindole-1,3,5-triones as key intermediates, which were obtained via Diels-Alder methodology using 1-aryl-3 siloxy-1, 3-butadienes as starting materials. Fischer indolization and aromatization processes afforded different methoxy-substituted arylpyrrolocarbazoles. The stereochemistry and conformation of the Diels-Alder products and the regiochemistry of the indolization reactions are supported by NMR and molecular modeling studies. PMID- 10843622 TI - The syntheses of pyrazino-containing sultines and their application in diels alder reactions with electron-poor olefins and AB - The Diels-Alder reactions of heterocyclic o-quinodimethanes, generated in situ from 6,7-disubstituted quinoxalino[2,3-d]-[1, 2lambda(4)]oxathiine 2-oxides (6a c), 2,3-disubstituted-8, 9-dihydro-6H-8lambda(4)-[1,2]oxathiino[4,5-g]quinoxalin 8-one (7a-c) (sultines), and pyrazinosultine (22), with electron-poor olefins and [60]fullerene are described. The heterocyclic-fused sultines 7a-c and 22 are readily prepared from the corresponding dibromides 9a-c and 24 with the commercially available Rongalite (sodium formaldehyde sulfoxylate). When heated in the presence of electron-poor dienophiles and [60]fullerene, all of the sultines underwent extrusion of SO(2), and the resulting heterocyclic o quinodimethanes (3a-d, 4a-c, and 25) were intercepted as the 1:1 adducts in good to excellent yields. The temperature-dependent (1)H NMR spectra of fullerene derivatives 31-38 show a dynamic process for the methylene protons. The activation free energies (DeltaG(c)()) determined for the boat-to-boat inversion of these pyrazino-containing C(60) compounds (31-34 and 38) are found to be in the range of 14.1-14.8 kcal/mol, but they are in the range of 15. 2 to >17.1 kcal/mol for adducts 35-37. The activation free energies (DeltaG(c)()) are significantly affected by (1) the orientations and (2) the substituents of the quinoxaline rings and (3) the extended benzannulation in the arenes of C(60) adducts (see Table 2), which implies that both electronic interactions and steric effects between the aromatic addends and C(60) are important. Tautomerization of methylquinoxaline to its enamine is invoked as a rationalization for the lowering of DeltaG(c)() in some of the fulleroadducts. PMID- 10843624 TI - Effects of medium on decarboxylation kinetics: 3-carboxybenzisoxazoles and their potential use as environmental probes in biochemistry. AB - The decarboxylation rate of the tetramethylguanidinium salt of 3-carboxy-6 nitrobenzisoxazole in 24 pure solvents and 36 dimethyl sulfoxide binary mixtures with diglyme, acetonitrile, benzene, dichloromethane, chloroform, and methanol was analyzed in the light of the SPP, SA, and SB pure solvent scales. The results allow one to rationalize the high sensitivity of this kinetics to the reaction medium and to assess the potential use of this compound as a probe in biochemical environments. The natural environment for comparison of this kinetics was found to be the gas phase rather than the aqueous medium. In the latter, the process is much faster owing to such high polarity, which, however, is strongly diminished by the high acidity of the medium. Based on our calculations, the rate constant for the decarboxylation kinetics in the gas phase must be in the region of 2 x 10(-10) s(-1) (i.e., 3 orders of magnitude smaller than in water). PMID- 10843623 TI - Palladium-catalyzed tert-butoxycarbonylation of trifluoroacetimidoyl iodides. AB - A modification and details of the palladium-catalyzed tert-butoxycarbonylation of 2,2,2-trifluoroacetimidoyl iodides 1, which gave the iminocarboxylates 2, one of the promising precursors to fluorinated alpha-amino acids, are described. The Pd catalyzed carbonylation reaction was remarkably promoted by the use of DMF or DMI as an additive, enough to achieve the selective formation of tert-butyl iminoesters. Nucleophilic alkylation of the imine moiety of 2 and subsequent removal of N- and O-protecting groups gave a variety of 2-substituted 2-amino 3,3,3-trifluoropropanoic acid derivatives 3 in high yields. PMID- 10843625 TI - Structural study of the solid-state photoaddition reaction of arylidenoxindoles AB - The photochemistry of isomeric 2-furyliden- and benzylidenoxindoles (2H-indol-2 ones) is examined. In solution E-Z isomerization is the only process via the excited singlet state (which fluoresces in glassy solution at 77 K and not at room temperature). In the crystalline state, the two (Z) derivatives are photostable, in accordance with the prediction based on the structural determination of the furylidene derivative, which adopts the unreactive Schmidt's gamma type arrangement. The (E) furylidene derivative (1a) gives efficiently (Phi = 0.3) the head-to-tail dimer, as indicated by the crystal structure, which is of the reactive alpha type, in full accord with the topochemical principles. In contrast, the corresponding benzylidene (1b) derivative reacts sluggishly (Phi < 0. 01) and mainly gives polymers, despite the fact that crystal structure determination shows that it likewise pertains to the alpha type and complies with the topochemical rules. The difference in reactivity is explained on the basis of (i) the twist of the phenyl ring with respect to the indole plane, and (ii) the higher overall cohesion energy and the lower interaction energy between facing molecules, as found from the charge density analysis for the crystals of 1b in comparison to those of 1a. This evidences a further stringent requirement for the occurrence of topochemical photodimerizations. PMID- 10843626 TI - Paterno-Buchi photocyclization of 2-siloxyfurans and carbonyl compounds. Notable substituent and carbonyl (Aldehyde vs ketone and singlet- vs triplet-excited state) effects on the regioselectivity (Double-bond selection) in the formation of bicyclic exo-oxetanes AB - Paterno-Buchi coupling, photochemical [2 + 2] cycloaddition, of carbonyl compounds 2a-f with 2-siloxyfurans 1a-d has been investigated in detail. The stereoselective formations of exo-oxetanes 3 and 4 were observed in high yields. The regioselectivity (double-bond selection, 3 vs 4) was found to be largely dependent upon the carbonyls, the substituents at the furan ring, and the excited state of the carbonyls (singlet vs triplet). The photoreaction of aldehydes 2a-c gave bicyclic exo-oxetanes 3 and 4 at regio-random, independent upon their excited states and the substituents at furan ring. However, the photoreaction of the triplet state of ketones 2d-f was found to give regioselectively exo-oxetanes 4, except for the 4-methyl-2-siloxyfurane 1d case. The singlet-excited state of acetone 2f gave both oxetanes 3 and 4 at regio-random. For the singlet-state photochemistry, the approach direction of the electrophilic oxygen of the excited carbonyls to the furan ring is proposed to be an important factor for the exo stereoselection. The Griesbeck model can rationalize the regio- and exo-selective formation of oxetanes in the triplet-state photoreaction. PMID- 10843627 TI - Studies toward the synthesis of pinolidoxin, a phytotoxic nonenolide from the fungus Ascochyta pinodes. Determination of the configuration at the C-7, C-8, and C-9 chiral centers and stereoselective synthesis of the C(6)-C(18) fragment. AB - The absolute stereochemistry at the C-7, C-8, and C-9 chiral centers of pinolidoxin (1) has been determined by chemical and spectral methods. First, the synthesis of four stereoisomeric fully benzoylated 2,3-erythro-1,2,3,4 heptanetetrols, corresponding to the C(6)-C(18) portion of the natural substance, has been accomplished starting from meso-tartaric acid. As next step, the selection of the synthetic tetrabenzoate possessing "natural" stereochemistry (10a'), suitable for absolute configuration determination, has been carried out by correlation with its "natural" homologue derived from degradation of pinolidoxin. Determination of the stereochemistry at the title chiral centers has been carried out by application of the Mosher's method both to 7a', a compound stereochemically related to 10a', and to pinolidoxin itself. The stereoselective synthesis of a protected form of the C(6)-C(18) portion of pinolidoxin, to be used in its total synthesis, has also been accomplished starting from commercially available D-erythronolactone. PMID- 10843628 TI - An unexpected carbon dioxide insertion in the reaction of trans-2, 4 disubstituted azetidine, trans-2,5-disubstituted pyrrolidine, or trans-2,6 disubstituted piperidine with diphenylthiophosphinic chloride and diphenylselenophosphinic chloride AB - In the reaction of trans-2,4-disubstituted azetidine, trans-2, 5-disubstituted pyrrolidine, or trans-2,6-disubstituted piperidine with diphenylthiophosphinic chloride or diphenylselenophosphinic chloride in acetonitrile in the presence of potassium carbonate at room temperature, an unexpected carbon dioxide insertion produced carbamic diphenylthiophosphinic or diphenylselenophosphinic anhydride in good yield. The same product could be also obtained when the reaction was carried out under carbon dioxide atmosphere using potassium hydroxide or triethylamine as a base. This is a very simple reaction process related to the fixation of carbon dioxide without a metal catalyst. PMID- 10843629 TI - Kinetics and mechanism of the oxygenation of potassium flavonolate. Evidence for an electron transfer mechanism. AB - The oxygenation of the potassium salt of flavonol (flaH) in absolute DMF leads to potassium O-benzoylsalicylate and carbon monoxide in 95% yield at 40 degrees C. Kinetic measurements resulted in the rate law -d[flaK]/dt = k(2)[flaK][O(2)]. The rate constant, activation enthalpy, and entropy at 313.16 K are as follows: k(2)/M(-)(1) s(-1) = (3.28 +/- 0.10) x 10(-1), DeltaH()/kJ mol(-1) = 29 +/- 2, DeltaS/J mol(-1) K(-1) = -161 +/- 6. The reaction fits a Hammett linear free energy relationship for 4'-substituted flavonols, and electron-releasing groups make the oxygenation reaction faster. The anodic oxidation wave potentials E(a) of the 4'-substituted flavonolates correlate well with reaction rates. At more negative E(a) values faster reaction rates were observed. EPR spectrum of the reaction mixture (g = 2.0038, dH = 1.8 G, a(H) = 0.9 G) showed the presence of flavonoxyl radical as a result of a SET from the flavonolate to dioxygen. PMID- 10843630 TI - Base-promoted isomerization of cis-4-formyl-2-azetidinones: chemoselective C4 epimerization vs rearrangement to cyclic enaminones. AB - Two simple, efficient, and complementary methods for the regiospecific C4 epimerization of cis-4-formyl-2-azetidinones 1-3 are described. The first method uses 40% aqueous dimethylamine as reagent in heterogeneous medium with benzene at room temperature, in the presence of benzyltributylammonium bromide (3-4 mol%) as the phase-transfer catalyst. This transformation tolerates alkyl, alkenyl, alkynyl, aryl, and alkoxy substituents at the C3 of the 2-azetidinone ring. However, limitations of this isomerization are as follows: (i) only N-(p methoxyphenyl)-beta-lactams can be used, and (ii) transformation is less compatible with heteroatomic substituents bonded to the C3 position of the 2 azetidinone ring. A highly general solution to these problems relies on the use of sodium carbonate as the isomerization reagent in different solvents. We also describe a novel base-promoted rearrangement of the beta-lactam ring to cyclic enaminones 6 and 21, involving an E1cB-elimination reaction in cis-4-formyl-2 azetidinones. PMID- 10843631 TI - Synthesis and novel reactivity of halomethyldimethylsulfonium salts AB - Iodomethyl-, chloromethyl-, and fluoromethyldimethylsulfonium salts, 4b-d, have been synthesized and are observed to be highly reactive molecules that exhibit extraordinary diversity with respect to the nature of their reactivity, undergoing facile direct substitution (S(N)2) reactions, but also being highly susceptible to electron-transfer reactions. Cyclic voltametry experiments indicated that the iodomethyldimethylsulfonium compound, 4b, is a potent electron acceptor, even surpassing the reactivity of perfluoro-n-alkyl iodides in that capacity. The iodo- and chloromethyldimethylsulfonium salts, 4b,c, as well as the analogous iodomethyltrimethylammonium salt, 3a, are shown to be reactive SET acceptors. PMID- 10843632 TI - Preparation of donor-sigma-acceptor molecules using arene-ruthenium chemistry. AB - Several donor-sigma acceptor (D-sigma-A) molecules with thioalkyl side chains have been prepared by ruthenium-activated nucleophilic aromatic substition (S(N)Ar) reactions. Selective substitution of chloride from cyclopentadienyl(1,4 dichlorobenzene)ruthenium by using piperazine derivatives as nucleophiles is addressed. This selectivity, in combination with further manipulation of the complexes, allows the preparation of unsymmetrically functionalized tetraalkyl-p phenylenediamine (TAPD) units which are difficult to synthesize by traditional organic S(N)Ar conditions. Phenanthroline-assisted decomplexation of the product arene-RuCp systems under UV irradiation is described. PMID- 10843633 TI - Toward an understanding of the selectivity in domino reactions. A DFT study of the reaction between acetylenedicarboxylic acid and 1, 3-Bis(2-furyl)propane AB - The mechanism of the domino reaction between acetylenedicarboxylic acid and 1,3 bis(2-furyl)propane has been theoretically studied in the framework of density functional theory. This domino process comprises two consecutive cycloaddition reactions: the first one is initialized by the nucleophilic attack of the C5 position of the furan ring to a conjugate position of acetylenedicarboxylic acid to give a zwitterionic intermediate, which by a subsequent ring-closure process affords an oxanorbornadiene intermediate. The second reaction is an intramolecular concerted cycloaddition of this intermediate to give the final dioxapentacyclic adduct. For the second cycloaddition, which corresponds to the step controlling the selectivity, eight alternative reaction pathways are found. Chemoselectivity, facial selectivity, and stereoselectivity of this domino reaction are related with the different approach modes of the tethered furan to the oxanorbornadiene system of the intermediate. The most favorable pathway takes place along an endo/syn approach of the furan ring relative to the bridged oxygen atom of the oxanorbornadiene system, with participation of the substituted double bond. An analysis of energetic contributions to the potential energy barriers for the intramolecular cycloadditions identifies the different factors controlling the reactive channels. Selectivity outcome is reproduced by these calculations. PMID- 10843634 TI - 3-Chloropropyl and 4-chlorobutyl phenyl ethers as sources of 1, 3-dilithiopropane and 1,4-dilithiobutane: sequential reaction with carbonyl compounds AB - The reaction of 3-chloropropyl and 4-chlorobutyl phenyl ethers (1) with lithium powder and a catalytic amount of DTBB (5% molar) in THF at -78 degrees C followed by successive treatment with a carbonyl compound [R(1)R(2)CO = Bu(t)CHO, Me(2)CO, (CH(2))(5)CO, (-)-menthone] at -78 to 20 degrees C and, after 1.5 h at this temperature, with a second one [R(3)R(4)CO = Bu(t)CHO, PhCHO, Me(2)CO, MeCOPr(n), (CH(2))(5)CO, (-)-menthone] at -78 degrees C leads, after hydrolysis with water, to the corresponding 1,5- and 1, 6-diols (2). Because of the competition of two different reductive cleavages, 1,4- and 1,5-diols 3 were also obtained as side reaction products. PMID- 10843635 TI - Photolysis of ((3-(Trimethylsilyl)propoxy)phenyl)phenyliodonium salts in the presence of 1-naphthol and 1-methoxynaphthalene AB - Direct photolysis of ((3-trimethylsilylpropoxy)phenyl)phenyliodonium salts with different counteranions (Cl(-), SbF(6)(-), and B(C(6)F(5))(4)(-)) in methanol leads to products by both heterolytic and homolytic processes. In the presence of 1-naphthol and 1-methoxynaphthalene, products formed by a heterolytic reaction disappear, suggesting an electron-transfer process occurs between excited 1 naphthol/1-methoxynaphthalene and the iodonium salts. In the case of 1 methoxynaphthalene, three phenylated methoxynaphthalene isomers are produced. These are produced as radical coupling products from the phenyl radical and 1 methoxynaphthalene radical cation. PMID- 10843637 TI - Oxa bowls: synthesis of new tetraoxa-cages and their C-H.O-mediated solid-state architecture AB - We have envisioned a new family of oxa-bowls, and a general approach to these entities has been proposed through polycyclic frameworks having two converging olefinic double bonds. Ozonolysis of such molecules was envisaged to give all-cis tetraaldehyde functionality, which could undergo intramolecular cascade acetalization to oxa-bowls having C(2)(v )()symmetry. Homohypostrophene 16a and its functionalized derivative 16b and hypostrophene 17 on ozonolysis furnished the oxa-cages 22a, 22b, and 23, respectively. Single-crystal X-ray analysis of 22a and 23 showed an interesting network of C-H.O interactions mainly involving the nonacidic cycloalkane hydrogen atoms. The molecule 22a exhibits an interesting tapelike C-H.O hydrogen bonding motif in its crystal structure. On the other hand, the molecule 23 revealed an undulated ribbonlike C-H. O motif. The "chain of rings" pattern of C-H.O interactions in 23 has been observed for the first time in the solid-state structure of oxa-bowls. PMID- 10843636 TI - Synthesis of chiral hydroxyl phospholanes from D-mannitol and their use in asymmetric catalytic reactions. AB - Chiral hydroxyl monophosphane 3 [(2S,3S,4S,5S)-3,4-dihydroxy-2, 5-dimethyl-1 phenylphospholane] and bisphospholanes 5a [1,2-bis[(2S, 3S,4S,5S)-3,4-dihydroxy 2,5-dimethylphospholanyl]benzene] and 5b [1, 2-bis[(2S,3S,4S,5S)-2,5-diethyl-3,4 dihydroxyphospholanyl]benzene] were synthesized from readily available D-mannitol in high yields. Strategies for protection and deprotection of OH-groups in the presence of phosphines have been explored. Rate acceleration in the Baylis Hillman reaction was observed when a hydroxyl phosphine was used as the catalyst. Rhodium complexes with chiral bisphospholanes are highly enantioselective catalysts for the asymmetric hydrogenation of various kinds of functionalized olefins such as dehydroamino acid derivatives, itaconic acid derivatives, and enamides. An interesting feature of the hydroxyl phospholane system is that hydrogenation of some substrates can be carried out in water with >99% ee and 100% conversion (e.g., itaconic acid). PMID- 10843638 TI - Enantioselective synthesis of rigid 2-aminotetralins. Utility of silicon as an oxygen and nitrogen surrogate in the tandem addition reaction. AB - Dimethylphenylsilyllithium undergoes a highly diastereoselective conjugate addition to chiral naphthyloxazoline 11. Electrophilic trapping of the resulting aza-enolate affords the tandem addition product (12) in high yields as a single diastereomer. The silicon, thus incorporated, may be protodesilylated and undergoes a Tamao oxidation to afford the corresponding alcohol. By chemical modification of the oxazoline, both the gamma-lactone (28) and the delta-lactone (37) were prepared. Reduction of each lactone followed by oxidation of the ensuing diol gave the keto aldehyde. Double reductive amination of the 1,4 dicarbonyl (from the gamma-lactone) allowed the synthesis of two novel hexahydrobenz[e]indoles, 20 and 35. Double reductive amination of the 1,5 dicarbonyl (from the delta-lactone) gave access to two novel octahydrobenzo[f]quinolines, 41 and 43. An unprecedented rearrangement of nitro alcohol 26 into lactone 28 is described and a reasonable mechanism for its formation postulated. PMID- 10843639 TI - Synthesis of tricyclic aromatic compounds by the intramolecular pauson-khand reaction promoted by molecular sieves(,)(1) AB - Pauson-Khand reactions are carried out with different substituted aromatic enynes, yielding tricyclic cyclopentenones related to natural products such as chromenes. Enynes are easily obtained in a two-step approximation from the corresponding salicylaldehydes. The reaction is promoted by dissolved TMANO (trimethylamine N-oxide) and/or 4 A molecular sieves. This new way of induction for the Pauson-Khand reaction increases yields remarkably, allowing the reaction of some substituted alkenes which fail to react in the absence of the zeolite. Isomerization of the double bond of the cyclopentenone ring is observed except when nonterminal triple bonds are used. For trisubstituted alkenes, an interrupted Pauson-Khand process is observed with moderate yields. PMID- 10843641 TI - Toward chiral polyhydroxylated dendrimers. Preparation and chiroptical properties AB - Four dendrimers (1b-4b) containing chiral vicinal diol-based subunits were prepared from their acetonide-protected precursors (1a-4a). The optical activity of these chiral dendritic structures was successfully modeled using structurally similar, low molecular weight model compounds. Using the [Phi](d) values of the low molecular weight model compounds 5b-7b, we calculated [Phi](d) values for dendrimers 1b-4b that agree to within 4.5% of the observed values. Agreement between the optical activity of the model compounds and that of the dendrimers leads to the conclusion that the conformational equilibria of the dendrimer subunits are not perturbed relative to those of the model compounds. PMID- 10843640 TI - Diastereoselective synthesis of hydantoin- and isoxazoline-substituted dispirocyclobutanoids. AB - Synthetic strategies for constructing novel achiral cyclobutanoid isoxazolidinoimidazolidinedione heterocycles, with a generalized structure of II, have been developed via 1,3-dipolar cycloaddition and carbanilide cyclization transformations from methylenecyclobutane 13. The exo methylene cyclobutane system has made the realization of some diasteroselectivity possible, such that the H-bond (Boc-NH) directed product (i.e., 14) was obtained with 3:1 selectivity relative to the non-H-bond directed product (i.e., 15). PMID- 10843642 TI - Synthesis and reactivity of trans-Tricyclo AB - The first synthesis of trans-tricyclo[4.2.0.0(1,3)]oct-4-ene (1), an ethenyl bridged spirohexane, was accomplished in four steps starting from Carpino et al. gem-dichloro ketone 6. An X-ray crystal structure of 1 with one substituent was obtained to provide geometry data on this novel ring system and to confirm the stereochemical assignment of the penultimate synthetic intermediate. Tricyclo[4.2.0. 0(1,3)]oct-4-ene is surprisingly stable. It reacts with glacial acetic acid but only slowly at 145 degrees C; the products were isolated and identified. A unimolecular rearrangement takes place at elevated temperatures (165 degrees C and higher), presumably, via a biradical intermediate to afford tricyclo[4.2.0.0(1,5)]oct-3-ene (23). The structure of this 1,5-bridged bicyclo[2.1.0]pentane derivative was established by NMR and an X-ray crystal structure of its Diels-Alder adduct with isobenzofuran. Tricyclo[4.2.0.0(1, 3)]oct-4-ene equilibrates with 23, so equilibrium constants and reaction rates were measured over a 20 degrees C temperature range from 180 degrees C to 200 degrees C. The difference in the heats of formation (DeltaDeltaH degrees (f) (23 1)) is -2.1 kcal/mol, which is in good agreement with ab initio (HF and MP2) calculations using the 6-31G(d) basis set (-1.9 (HF) and -1.4 (MP2) kcal/mol). Computations on trans-tricyclo[4.2.0.0(1,3)]octane and spirohexane also were carried out, and the structures and energies were compared. PMID- 10843643 TI - Stereospecific total syntheses of sphingosine and its analogues from L-serine. PMID- 10843645 TI - Synthesis of gamma,delta-didehydrohomoglutamates by the phosphine-catalyzed gamma addition reaction to acetylenic esters PMID- 10843644 TI - On the synthesis and NMR analysis of tetrabutylammonium triphenyldifluorosilicate. PMID- 10843646 TI - Aryltitanium species through the reaction of N,N-dialkylarylamines with TiCl(4): oxidative coupling, N-dealkylation, and reaction with electrophiles PMID- 10843647 TI - A short and efficient enantiospecific synthesis of (+)-(2R, 6S)-cis-gamma-irone via a highly diastereoselective protonation. PMID- 10843648 TI - A practical and facile route for the preparation of 18-norandrostan-17-ones from androstan-17-ones using SmI(2)-promoted cyclization and dehydroxylation. PMID- 10843650 TI - Conformational studies by dynamic NMR. 76.(1) stereodynamics of ring inversion of Bicyclo PMID- 10843649 TI - Daphnezomines F and G: novel alkaloids with 1-azabicyclo[5.2. 2]undecane moiety from Daphniphyllum humile. PMID- 10843651 TI - Total synthesis of (+/-)-nudenoic acid. PMID- 10843652 TI - A novel heterobicyclic framework by successive ene reactions PMID- 10843655 TI - Risperidone and Cognitive Functioning in Elderly Schizophrenic Patients(Dr. Madhusoodanan comments). PMID- 10843653 TI - Stereodivergent synthesis of (E)- and (Z)-2-alken-4-yn-1-ols from 2-propynoic acid: A practical route via 2-alken-4-ynoates PMID- 10843656 TI - Cutting edge: CD46, a new costimulatory molecule for T cells, that induces p120CBL and LAT phosphorylation. AB - The widely expressed transmembrane molecule CD46 is the complement regulatory receptor for C3b as well as the receptor for several pathogens. Beside its binding functions, CD46 is also able to transduce signals. We showed that CD46 aggregation on human T cells induces p120CBL and linker for activation of T cells (LAT) phosphorylation. These two proteins are adaptor proteins known to regulate TCR signaling. p120CBL is a complex adaptor protein involved in negatively regulating signaling events, whereas LAT is a transmembrane adaptor protein found in glycolipid-enriched microdomains essential for T cell activation. Therefore, we investigated if a CD46/TCR costimulation would affect T cell activation. Indeed, CD46/CD3 costimulation strongly promotes T cell proliferation. Therefore, we propose that CD46 acts as a potent costimulatory molecule for human T cells. PMID- 10843657 TI - Cutting edge: T cell-mediated pathology in murine Lyme borreliosis. AB - Even in the absence of an appropriate model or direct evidence, T cells have been hypothesized to exacerbate the manifestations of Lyme disease. To define definitely the role of T cells in Lyme disease, the course of disease in immunocompetent and B cell-deficient mice was compared. By 8 wk postinoculation, immunocompetent mice resolved both carditis and arthritis, whereas foci of myocarditis and severe destructive arthritis characterized disease of B cell deficient mice. Cell transfer experiments using infected B6-Rag1 knock out mice demonstrated that: 1) innate immunity mediated the initial sequelae of infection, 2) transferring both naive T cells and B cells induced resolution of carditis and arthritis, 3) infected mice reconstituted with T cells developed myocarditis and severe destructive arthritis, and 4) CD4+ T cells were responsible for the observed immune-mediated pathology. These data demonstrate directly the deleterious effect of T cells in Lyme disease. PMID- 10843658 TI - Cutting edge: soluble HLA-G1 triggers CD95/CD95 ligand-mediated apoptosis in activated CD8+ cells by interacting with CD8. AB - The nonpolymorphic soluble HLA-G1 (sHLA-G1) isoform has been reported to be secreted by trophoblast cells at the materno-fetal interface, suggesting that it may act as immunomodulator during pregnancy. In this paper, we report that affinity-purified beta2-microglobulin-associated sHLA-G1 triggered apoptosis in activated, but not resting CD8+ peripheral blood cells. We demonstrate by Western blotting that sHLA-G1 enhanced CD95 ligand expression in activated CD8+ cells. Cytotoxicity was inhibited by preincubation of the cells with a CD95 antagonist mAb (ZB4) or a soluble recombinant CD95-Fc, indicating that apoptosis is mediated through the CD95/CD95 ligand pathway. Finally, we show that such sHLA-G1-induced apoptosis depends on the interaction with CD8 molecules, with cell death being blocked by various CD8 mAbs. PMID- 10843659 TI - Altered composition of the immunological synapse in an anergic, age-dependent memory T cell subset. AB - In young mice, memory CD4 T lymphocytes with high P-glycoprotein activity (P gp(high)) are unresponsive to TCR stimulation in vitro but can be activated by PMA plus ionomycin. The proportion of these hyporesponsive cells increases considerably with age. The earliest events in T cell activation were studied in P gp(high) and P-gp(low) CD4 memory cells at the single-cell level using confocal immunofluorescence methods. Recruitment of both linker for activation of T cells (LAT) and protein kinase C-theta to the immunological synapse, i.e., the site of T cell interaction with stimulator cells, was greatly impaired in P-gp(high) cells from both young and old mice. Translocation of NF-AT to the nucleus, CD69 expression, and proliferative capacity were also diminished to a similar extent in P-gp(high) cells under the same activation conditions. In contrast, movement of c-Cbl to the synapse region occurred in a high proportion of CD4 memory T cells regardless of P-gp subset or age. Moreover, although P-gp(low) cells frequently recruited both c-Cbl and LAT to the APC synapse, cells in the less responsive P-gp(high) subset frequently relocated c-Cbl, but not LAT, to the interface region. In some systems, c-Cbl can act as a negative regulator of receptor-dependent tyrosine kinases, and alterations of c-Cbl to LAT ratios in the P-gp(high) subset may thus contribute to the hyporesponsiveness of this age dependent, anergic memory cell population. PMID- 10843660 TI - Immune complex and Fc receptor-mediated augmentation of antigen presentation for in vivo Th cell responses. AB - It has recently been established that FcRs are involved in the triggering of type II and III inflammatory responses. Although FcR is not believed to be involved in the regulation of T cell function, the in vivo contribution of FcRs to T cell function still remains unclear. We analyzed in vivo responses of delayed-type hypersensitivity and proliferation of CD4+ T cells to Ags in FcRgamma-/- mice lacking the expression and function of FcgammaRI, FcgammaRIII, and FcepsilonRI. We found that the delayed-type hypersensitivity response in FcRgamma-/- mice is significantly decreased compared with that in wild-type mice. Moreover, the secondary responses of proliferation and cytokine production as well as the Ab formation by CD4+ T cells from FcRgamma-/- mice to Ag and normal APCs were also reduced. In contrast, in vitro primary T cell proliferative responses upon stimulation with anti-TCR Ab or MLR as well as in vivo primary response against staphylococcus enterotoxin B administration were not different between T cells from FcRgamma-/- and wild-type mice. In addition, the Ag presentation function of APCs from unimmunized FcRgamma-/- mice was normal. On the other hand, Ab deficient mice also revealed impaired T cell responses. These results demonstrate that the defective T cell responses in FcRgamma-/- mice were due to impaired Ag presentation during in vivo priming not to a defect in T cells. Therefore, they suggest that the FcRs on APCs mediate efficient priming of Th cell responses in vivo in an immune complex-dependent manner. PMID- 10843661 TI - Immunodominance among EBV-derived epitopes restricted by HLA-B27 does not correlate with epitope abundance in EBV-transformed B-lymphoblastoid cell lines. AB - Using synthetic peptides, the HLA-B27-restricted CTL response to EBV in asymptomatic virus carriers has been mapped to four epitope regions in EBV latent cycle Ags. One of these peptide-defined epitopes (RRIYDLIEL) tends to be immunodominant and is recognized in the context of all three B27 subtypes studied, B*2702, B*2704, and B*2705. The other peptide-defined epitopes induce responses only in the context of one subtype, the immunogenic combinations being RRARSLSAERY/B*2702, RRRWRRLTV/B*2704, and FRKAQIQGL/B*2705. We used immunoaffinity chromatography to isolate the naturally presented viral peptides associated with these MHC class I molecules on the surface of EBV-transformed B LCL. Using CTL reconstitution assays in conjunction with mass spectrometry, we established that the naturally processed and presented peptides are identical with the previously identified synthetic sequences. Despite the subtype-specific immunogenicity of three of the four epitopes, all four epitope peptides were found in association with each of the three different HLA-B27 subtypes. Indeed, those peptides that failed to induce a response in the context of a particular HLA-B27 subtype were frequently presented at greater abundance by that subtype than were the immunogenic peptides. Furthermore, among the peptides that did induce a response, immunodominance did not correlate with epitope abundance; in fact the immunodominant RRIYDLIEL epitope was least abundant, being present at less than one copy per cell. The relationship of this unexpected finding to the persistence of EBV is discussed. PMID- 10843662 TI - Inhibition of human NK cell-mediated killing by CD1 molecules. AB - It is now well established that NK cells recognize classical and nonclassical MHC class I molecules and that such recognition typically results in the inhibition of target cell lysis. Given the known structural similarities between MHC class I and non-MHC-encoded CD1 molecules, we investigated the possibility that human CD1a, -b, and -c proteins might also function as specific target structures for NK cell receptors. Here we report that expression of CD1a, -b, or -c can partially inhibits target cell lysis by freshly isolated human NK cells and cultured NK lines. The inhibitory effects of CD1 molecules on NK cell could be shown upon expression of individual CD1 proteins in transfected NK-sensitive target cells, and these effects could be reversed by incubation of the target cells with mAbs specific for the expressed form of CD1. Inhibitory effects of CD1 expression on NK-mediated lysis could also be shown for cultured human dendritic cells, which represent a cell type that prominently expresses the various CD1 proteins in vivo. In addition, the bacterial glycolipid Ags known to be bound and presented by CD1 proteins could significantly augment the observed inhibitory effects on target cell lysis by NK cells. PMID- 10843663 TI - Analysis of soluble and cell surface factors regulating anti-DNA topoisomerase I autoantibody production demonstrates synergy between Th1 and Th2 autoreactive T cells. AB - The cellular and subcellular events governing Ab production with specificity for self Ags are poorly understood. In this study we examined the role of cellular interactions and cytokines in regulating the production of anti-DNA topoisomerase I (topo I) Ab, a major autoantibody in patients with systemic sclerosis (SSc). Topo I-specific T cell clones derived from SSc subjects and healthy donors were cultured with autologous peripheral blood B cells. Anti-topo I Ab production was induced by five of seven topo I-specific T cell clones derived from SSc subjects, but by none of eight T cell clones generated from healthy controls. However, two of the T cell clones from healthy controls provided help to HLA-DR-matched SSc B cells to produce anti-topo I Ab. The analysis of cytokine mRNA expression revealed that the ability to promote anti-topo I autoantibody production was strictly correlated with IL-2 and IL-6 expression by the T cell clones. Kinetic studies showed that IL-2 was required throughout the culture period for maximal autoantibody production and that both MHC-TCR and CD40-CD40L interactions were essential during the early phase of the culture. IL-6 was important in the late phase. Th1 clones (producing IL-2, but no IL-6) and Th2 clones (producing IL-6, but no IL-2) synergically activated autologous B cells to produce anti-topo I Ab. These results indicate that T cell-dependent B cell activation resulting in anti topo I autoantibody production requires a series of temporally defined cell contact and soluble stimuli. PMID- 10843664 TI - The selective proteasome inhibitors lactacystin and epoxomicin can be used to either up- or down-regulate antigen presentation at nontoxic doses. AB - The complete inhibition of proteasome activities interferes with the production of most MHC class I peptide ligands as well as with cellular proliferation and survival. In this study we have investigated how partial and selective inhibition of the chymotrypsin-like activity of the proteasome by the proteasome inhibitors lactacystin or epoxomicin would affect Ag presentation. At 0.5-1 microM lactacystin, the presentation of the lymphocytic choriomeningitis virus-derived epitopes NP118 and GP33 and the mouse CMV epitope pp89-168 were reduced and were further diminished in a dose-dependent manner with increasing concentrations. Presentation of the lymphocytic choriomeningitis virus-derived epitope GP276, in contrast, was markedly enhanced at low, but abrogated at higher, concentrations of either lactacystin or epoxomicin. The inhibitor-mediated effects were thus epitope specific and did not correlate with the degradation rates of the involved viral proteins. Although neither apoptosis induction nor interference with cellular proliferation was observed at 0.5-1 microM lactacystin in vivo, this concentration was sufficient to alter the fragmentation of polypeptides by the 20S proteasome in vitro. Our results indicate that partial and selective inhibition of proteasome activity in vivo is a valid approach to modulate Ag presentation, with potential applications for the treatment of autoimmune diseases and the prevention of transplant rejection. PMID- 10843665 TI - The thyrotropin (thyroid-stimulating hormone) receptor is expressed on murine dendritic cells and on a subset of CD45RBhigh lymph node T cells: functional role for thyroid-stimulating hormone during immune activation. AB - Thyroid-stimulating hormone (TSH), a central neuroendocrine mediator of the hypothalamus-pituitary-thyroid axis, has been shown to affect various aspects of immunological development and function. To gain a better understanding of TSH involvement within the mammalian immune system, the expression and distribution of the TSH receptor (TSHr) has been studied by immunoprecipitation and by flow cytometric analyses. Using highly enriched populations of B cells, T cells, and dendritic cells, trace amounts of TSHr were precipitated from B cells and T cells, whereas high levels of TSHr were precipitated from the dendritic cell fraction. Flow cytometric analyses of TSHr expression on splenic and lymph node T cells revealed a major difference between those tissues in that only 2-3% of splenic T cells were TSHr+, whereas 10-20% of CD4+8- and CD4-8+ lymph node T cells expressed the TSHr, which was exclusively associated with CD45RB(high) cells and was not expressed during or after activation. The TSHr was not present on cells of the immune system during fetal or neonatal life. However, recombinant TSHbeta was found to significantly enhance the phagocytic activity of dendritic cells from adult animals and to selectively augment IL-1beta and IL-12 cytokine responses of dendritic cells following phagocytic activation. These findings identify a novel immune-endocrine bridge associated with professional APCs and naive T cells. PMID- 10843666 TI - M-1/M-2 macrophages and the Th1/Th2 paradigm. AB - Evidence is provided that macrophages can make M-1 or M-2 responses. The concept of M-1/M-2 fomented from observations that macrophages from prototypical Th1 strains (C57BL/6, B10D2) are more easily activated to produce NO with either IFN gamma or LPS than macrophages from Th2 strains (BALB/c, DBA/2). In marked contrast, LPS stimulates Th2, but not Th1, macrophages to increase arginine metabolism to ornithine. Thus, M-1/M-2 does not simply describe activated or unactivated macrophages, but cells expressing distinct metabolic programs. Because NO inhibits cell division, while ornithine can stimulate cell division (via polyamines), these results also indicate that M-1 and M-2 responses can influence inflammatory reactions in opposite ways. Macrophage TGF-beta1, which inhibits inducible NO synthase and stimulates arginase, appears to play an important role in regulating the balance between M-1 and M-2. M-1/M-2 phenotypes are independent of T or B lymphocytes because C57BL/6 and BALB/c NUDE or SCID macrophages also exhibit M-1/M-2. Indeed, M-1/M-2 proclivities are magnified in NUDE and SCID mice. Finally, C57BL/6 SCID macrophages cause CB6F1 lymphocytes to increase IFN-gamma production, while BALB/c SCID macrophages increase TGF-beta production. Together, the results indicate that M-1- or M-2-dominant macrophage responses can influence whether Th1/Th2 or other types of inflammatory responses occur. PMID- 10843667 TI - Monocyte chemoattractant protein-1 enhances gene expression and synthesis of matrix metalloproteinase-1 in human fibroblasts by an autocrine IL-1 alpha loop. AB - Monocyte chemoattractant protein-1 (MCP-1), a member of the C-C chemokine superfamily, has recently been shown to be involved in the pathogenesis of tissue fibrosis. In vitro studies demonstrated that MCP-1 up-regulates type I collagen gene expression via endogenous production of TGF-beta in rat lung fibroblasts. We here show that recombinant human MCP-1 affects gene expression of interstitial collagenase (matrix metalloproteinase-1 (MMP-1)) in primary human skin fibroblasts and a stable fibroblast cell line. MMP-1 mRNA was induced by MCP-1 (10 ng/ml) as early as 6 h and reached a maximal expression at 24 h. MCP-1 also caused an increase of MMP-2 mRNA expression in both types of fibroblasts at 48 h. Interestingly, tissue inhibitor of metalloproteinase-1 (TIMP-1) mRNA was also up regulated by MCP-1, and TIMP-1 mRNA expression peaked at 48 h in both types of fibroblasts. Immunoblot analysis demonstrated increased levels of MMP-1 and TIMP 1 protein in the culture supernatants of primary fibroblasts stimulated with MCP 1. In addition, MCP-1 strongly induced IL-1 alpha mRNA expression in dermal fibroblasts in parallel with the induction of MMP-1. Preincubation with IL-1 receptor antagonist almost completely abrogated the expression of MMP-1 mRNA, and partially inhibited MMP-1 synthesis induced by MCP-1. Transient transfection of primary skin fibroblasts with a MMP-1 promoter-reporter construct indicated a dose-dependent increase in promoter activity by MCP-1 stimulation. These data demonstrate that MCP-1 up-regulates MMP-1 mRNA expression and synthesis in human skin fibroblasts at a transcriptional level and provide evidence that this is mediated by an IL-1 alpha autocrine loop. PMID- 10843668 TI - TNF-alpha-induced secretion of C-C chemokines modulates C-C chemokine receptor 5 expression on peripheral blood lymphocytes. AB - Peripheral blood lymphocytes express CCR5, a chemokine receptor for immune cell migration and calcium signaling that serves as an important coreceptor for the HIV. After in vitro stimulation, CCR5 expression is dramatically increased on mature T lymphocytes, especially on the CD45RO+ memory subset. In this study, we report that TNF-alpha delays the surface expression of CCR5 on PBLs after activation and diminishes CCR5 irrespective of its initial level. Functional loss of CCR5 is reflected in a decreased capability of the treated cells to migrate and signal calcium after MIP-1beta stimulation. The effect is mediated via the p80 type II TNF receptor (TNFR2), which induces NF-kappaB among other factors, leading to an enhanced secretion of the chemokines macrophage-inflammatory protein-1alpha, macrophage-inflammatory protein-1beta, and RANTES. Expression of these chemokines directly down-regulates CCR5. These findings reveal a new regulatory mechanism utilized by activated peripheral T cells to modulate their chemotaxis and potentially other functions mediated by CCR5, including the infection of T lymphocytes by macrophage-tropic HIV strains. PMID- 10843669 TI - Proliferating cell nuclear antigen as the cell cycle sensor for an HLA-derived peptide blocking T cell proliferation. AB - Synthetic peptides corresponding to structural regions of HLA molecules are novel immunosuppressive agents. A peptide corresponding to residues 65-79 of the alpha chain of HLA-DQA03011 (DQ65-79) blocks cell cycle progression from early G1 to the G1 restriction point, which inhibits cyclin-dependent kinase-2 activity and phosphorylation of the retinoblastoma protein. A yeast two-hybrid screen identified proliferating cell nuclear Ag (PCNA) as a cellular ligand for this peptide, whose interaction with PCNA was further confirmed by in vitro biochemistry. Electron microscopy demonstrates that the DQ65-79 peptide enters the cell and colocalizes with PCNA in the T cell nucleus in vivo. Binding of the DQ65-79 peptide to PCNA did not block polymerase delta (pol delta)-dependent DNA replication in vitro. These findings support a key role for PCNA as a sensor of cell cycle progression and reveal an unanticipated function for conserved regions of HLA molecules. PMID- 10843670 TI - Reverse signaling through transmembrane TNF confers resistance to lipopolysaccharide in human monocytes and macrophages. AB - We have previously reported that the CD14+ monocytic subpopulation of human PBMC induces programmed cell death (apoptosis) in cocultured endothelial cells (EC) when stimulated by bacterial endotoxin (LPS). Apoptosis is mediated by two routes, first via transmembrane TNF-alpha (mTNF) expressed on PBMC and, in addition, by TNF-independent soluble factors that trigger apoptosis in EC. Neutralizing anti-TNF mAb completely blocked coculture-mediated apoptosis, despite the fact that there should have been additional soluble cell death factors. This led to the hypothesis that a reverse signal is transmitted from the TNF receptor on EC to monocytes (MO) via mTNF that prevents the production of soluble apoptotic factors. Here we have tested this hypothesis. The results support the idea of a bidirectional cross-talk between MO and EC. Peripheral blood MO, MO-derived macrophages (MPhi), or the monocytic cell line Mono Mac 6 were preincubated with human microvascular EC that constitutively express TNF receptor type I (TNF-R1) and subsequently stimulated with LPS. Cell-free supernatants of these preparations no longer induced EC apoptosis. The preincubation of MO/MPhi with TNF-reactive agents, such as mAb and soluble receptors, also blocked the production of death factors, providing further evidence for reverse signaling via mTNF. Finally, we show that reverse signaling through mTNF mediated LPS resistance in MO/MPhi as indicated by the down regulation of LPS-induced soluble TNF and IL-6 as well as IL-1 and IL-10. PMID- 10843671 TI - Transcription-coupled and global genome repair differentially influence UV-B induced acute skin effects and systemic immunosuppression. AB - Exposure to UV-B radiation impairs immune responses in mammals by inhibiting especially Th1-mediated contact hypersensitivity and delayed-type hypersensitivity. Immunomodulation is not restricted to the exposed skin, but is also observed at distant sites, indicating the existence of mediating factors such as products from exposed skin cells or photoactivated factors present in the superficial layers. DNA damage appears to play a key role, because enhanced nucleotide excision repair (NER) strongly counteracts immunosuppression. To determine the effects of the type and genomic location of UV-induced DNA damage on immunosuppression and acute skin reactions (edema and erythema) four congenic mouse strains carrying different defects in NER were compared: CSB and XPC mice lacking transcription-coupled or global genome NER, respectively, as well as XPA and TTD/XPD mice carrying complete or partial defects in both NER subpathways, respectively. The major conclusions are that 1) transcription-coupled DNA repair is the dominant determinant in protection against acute skin effects; 2) systemic immunomodulation is only affected when both NER subpathways are compromised; and 3) sunburn is not related to UV-B-induced immunosuppression. PMID- 10843672 TI - Sustained expression of CD154 (CD40L) and proinflammatory cytokine production by alloantigen-stimulated umbilical cord blood T cells. AB - Recent data suggests that graft-versus-host disease (GVHD) is initiated by host APCs. Blockade of CD40:CD154 interactions between APCs and T cells in vivo induces T cell tolerance to host alloantigen and dramatically reduces GVHD. Because allogeneic cord blood (CB) transplantation results in a lower incidence and severity of acute GVHD compared with bone marrow transplantation, we have investigated whether CB T cells can express CD154 in response to stimulation by allogeneic monocyte-derived dendritic cells (MDDC) and have used 5- (and 6 )carboxyfluorescein diacetate succinimidyl ester (CFSE) labeling in combination with intracellular cytokine analysis to assess the proliferation and cytokine profiles of alloantigen-responsive cells. CB T cells stimulated with allogeneic MDDC showed stronger proliferation than adult blood T cells. Surface CD154 expression was detected in the actively dividing CFSElow populations of both the CD4+ and CD4- subsets and was brightest in cells that had divided the most. Assessment of supernatants from MDDC-stimulated CB and adult blood T cells showed no significant difference in the levels of either IFN-gamma or TNF-alpha, but CB T cell supernatants did show a significant lack of detectable IL-2. Intracellular cytokine analysis revealed that dividing CB T cells had been primed to produce IFN-gamma, TNF-alpha, and IL-2 on restimulation. Further phenotype analysis showed that 75% of CB T cells producing IFN-gamma were CD8+. These data suggest that MDDC-stimulated CB T cells express functional CD154 and provide enough costimulation for dendritic cells to prime naive CD8+ CB T cells and induce type 1 cytokine production. PMID- 10843673 TI - Glucocorticoids regulate TCR-induced elevation of CD4: functional implications. AB - CD4 serves as a coreceptor during Ag recognition by the TCR. This interaction results in a marked increase in the sensitivity of a T cell to Ag presented by MHC class II molecules. Here we report that activation of T cells either by plate bound mAb (anti-TCR, anti-CD3) or soluble activators (staphylococcal enterotoxin A, Con A) is associated with an (up to 3-fold) increase in CD4 cell surface expression on CD25+ cells, which was maximal after 72-96 h. Incubation with the glucocorticoid hormone corticosterone (CORT) shifted the enhancement of CD4 expression to a point about 24 h earlier than that observed in control cultures. In parallel, the proliferative response of these CORT-treated cells was profoundly enhanced. An involvement of increased CD4 expression in this enhanced proliferative response was evidenced by the observation that T cell proliferation in CORT-treated cultures was much less sensitive to inhibition by an inhibitory, nondepleting anti-CD4 mAb than that in control cultures. TCR down-regulation was, however, not affected by CORT. Thus, based on this study and previous reports we propose that both TCR-mediated signals and glucocorticoids are important physiological regulators of CD4 expression. In addition, these findings may be of significance for the sensitivity of CD4+ cells to HIV infection upon T cell activation, as the efficacy of primary patient HIV entry depends on the level of surface CD4. PMID- 10843674 TI - Analysis of the diversity of a sheep antibody repertoire as revealed from a bacteriophage display library. AB - We have applied bacteriophage display technology to construct and analyze the diversity of an IgG library of >1 x 108 clones from an adult sheep immunized against the hapten atrazine. We have identified eight new VH gene families (VH2 VH9) and five new Vkappa gene families (VkappaV-VkappaIX). The heavy and kappa light chain variable region gene loci were found to be far more diverse than previously thought. PMID- 10843675 TI - Melphalan and other anticancer modalities up-regulate B7-1 gene expression in tumor cells. AB - In this study, we show that administration of low-dose melphalan (l -PAM, l phenylalanine mustard) to mice bearing a large MOPC-315 plasmacytoma led to a rapid up-regulation of B7-1 (CD80), but not B7-2 (CD86), expression on the surface of MOPC-315 tumor cells. This l -PAM-induced preferential up-regulation of B7-1 surface expression was due, at least in part, to a direct effect of l PAM on the tumor cells, as in vitro exposure of MOPC-315 tumor cells to l -PAM led to the preferential up-regulation of B7-1 surface expression. Moreover, in vitro exposure of MOPC-315 tumor cells to two other anticancer modalities, gamma irradiation and mitomycin C, resulted in the preferential up-regulation of B7-1 surface expression. This effect was not restricted to MOPC-315 tumor cells, as preferential up-regulation of B7-1 surface expression was observed also following in vitro exposure of the P815 mastocytoma (that is negative for both B7-1 and B7 2 surface expression) to any of the three anticancer modalities. The up regulation of B7-1 surface expression following in vitro exposure of tumor cells to l -PAM, gamma-irradiation, or mitomycin C required de novo protein and RNA synthesis, and was associated with the accumulation of mRNA for B7-1 within 4-8 h, indicating that the regulation of B7-1 expression is at the RNA transcriptional level. These results have important implications for an additional immune-potentiating mechanism of these anticancer modalities in clinical setting. PMID- 10843676 TI - Gender-dependent IL-12 secretion by APC is regulated by IL-10. AB - Female SJL mice preferentially mount Th1-immune responses and are susceptible to the active induction of experimental allergic encephalomyelitis. By contrast, young adult male SJL are resistant to experimental allergic encephalomyelitis due to an APC-dependent induction of Th2 cells. The basis for this gender-dependent differential T cell induction was examined by analysis of macrophage APC cytokine secretion during T cell activation. APC derived from females secrete IL-12, but not IL-10, during T cell activation. By contrast, APC derived from males secrete IL-10, but not IL-12, during T cell activation. Activation of T cells with APC derived from the opposite sex demonstrated that these cytokines were derived from the respective APC populations. Furthermore, inhibition of IL-10, but not TGF beta, during T cell activation resulted in the secretion of IL-12 by male-derived APC. APC from naive male mice, in which IL-10 was reduced in vivo before isolation, also secrete IL-12, demonstrating altered APC cytokine secretion was due to an environment high in IL-10 before Ag encounter. Finally, APC derived from castrated male mice preferentially secrete IL-12 during T cell activation. These data demonstrate a link between gonadal hormones and APC activity and suggest that these hormones alter the APC, thereby influencing cytokine secretion during initial T cell activation. PMID- 10843678 TI - Competition for specific intrathymic ligands limits positive selection in a TCR transgenic model of CD4+ T cell development. AB - Efficient positive selection of a broad repertoire of T cells is dependent on the presentation of a diverse array of endogenous peptides on MHC molecules in the thymus. It is unclear, however, whether the development of individual TCR specificities is influenced by the abundance of their selecting ligands. To examine this, we analyzed positive selection in a transgenic mouse carrying a TCR specific for the human CLIP:I-Ab class II complex. We found that these mice exhibit significantly reduced CD4+ T cell development compared with two other transgenic mice carrying TCRs selected on I-Ab. Moreover, many of the selected cells in these mice express endogenous and transgenic receptors as a consequence of dual TCRalpha expression. Dramatic enhancement of the selection efficiency is observed, however, when fewer transgenic cells populate the thymus in mixed bone marrow chimeras. These results suggest that positive selection is limited by the availability of selecting peptides in the thymus. This becomes apparent when large numbers of thymocytes compete for such peptides in TCR transgenic animals. Under such conditions, thymocytes appear to undergo further TCRalpha gene rearrangement to produce a receptor that may be selected more efficiently by other thymic self-peptides. PMID- 10843677 TI - IL-2 activation of NK cells: involvement of MKK1/2/ERK but not p38 kinase pathway. AB - IL-2 stimulates extracellular signal-regulated protein kinase (ERK) and p38 mitogen-activated protein kinase (MAPK) in various immune cell populations. The functional roles that these kinases play are still unclear. In this study, we examined whether MAPK kinase (MKK)/ERK and p38 MAPK pathways are necessary for IL 2 to activate NK cells. Using freshly isolated human NK cells, we established that an intact MKK/ERK pathway is necessary for IL-2 to activate NK cells to express at least four known biological responses: LAK generation, IFN-gamma secretion, and CD25 and CD69 expression. IL-2 induced ERK activation within 5 min. Treatment of NK cells with a specific inhibitor of MKK1/2, PD98059, during the IL-2 stimulation blocked in a dose-dependent manner each of four sequelae, with inhibition of lymphokine-activated killing induction being least sensitive to MKK/ERK pathway blockade. Activation of p38 MAPK by IL-2 was not detected in NK cells. In contrast to what was observed by others in T lymphocytes, SB203850, a specific inhibitor of p38 MAPK, did not inhibit IL-2-activated NK functions. This data indicate that p38 MAPK activation was not required for IL-2 to activate NK cells for the four functions examined. These results reveal selective signaling differences between NK cells and T lymphocytes; in NK cells, the MKK/ERK pathway and not p38 MAPK plays a critical positive regulatory role during activation by IL-2. PMID- 10843679 TI - Analysis of the human neonatal thymus: evidence for a transient thymic involution. AB - The neonatal period is marked by the impairment of the major components of both innate and adaptive immunity. We report a severe depletion of cortical CD4+CD8+ double-positive thymocytes in the human neonatal thymus. This drastic reduction in immature double-positive cells, largely provoked by an increased rate of cell death, could be observed as early as 1 day after birth, delaying the recovery of the normal proportion of this thymocyte subset until the end of the first month of postnatal life. Serum cortisol levels were not increased in newborn donors, indicating that the neonatal thymic involution is a physiological rather than a stress-associated pathological event occurring in the perinatal period. Newborn thymuses also showed increased proportions of both primitive CD34+CD1- precursor cells and mature TCRalphabetahighCD69-CD1-CD45RO+/RAdull and CD45ROdull/RA+ cells, which presumably correspond to recirculating T lymphocytes into the thymus. A notable reinforcement of the subcapsular epithelial cell layer as well as an increase in the intralobular extracellular matrix network accompanied modifications in the thymocyte population. Additionally neonatal thymic dendritic cells were found to be more effective than dendritic cells isolated from children's thymuses at stimulating proliferative responses in allogeneic T cells. All these findings can account for several alterations affecting the peripheral pool of T lymphocytes in the perinatal period. PMID- 10843680 TI - Fc gamma receptor IIB on follicular dendritic cells regulates the B cell recall response. AB - Generation of the B cell recall response appears to involve interaction of Ag, in the form of an immune complex (IC) trapped on follicular dendritic cells (FDCs), with germinal center (GC) B cells. Thus, the expression of receptors on FDC and B cells that interact with ICs could be critical to the induction of an optimal recall response. FDCs in GCs, but not in primary follicles, express high levels of the IgG Fc receptor Fc gamma RIIB. This regulated expression of Fc gamma RIIB on FDC and its relation to recall Ab responses were examined both in vitro and in vivo. Trapping of IC in spleen and lymph nodes of Fc gamma RII-/- mice was significantly reduced compared with that in wild-type controls. Addition of ICs to cultures of Ag-specific T and B cells elicited pronounced Ab responses only in the presence of FDCs. However, FDCs derived from Fc gamma RIIB-/- mice supported only low level Ab production in this situation. Similarly, when Fc gamma RIIB-/- mice were transplanted with wild-type Ag-specific T and B cells and challenged with specific Ag, the recall responses were significantly depressed compared with those of controls with wild-type FDC. These results substantiate the hypothesis that FcgammaRIIB expression on FDCs in GCs is important for FDCs to retain ICs and to mediate the conversion of ICs to a highly immunogenic form and for the generation of strong recall responses. PMID- 10843681 TI - The ADP-ribosylating CTA1-DD adjuvant enhances T cell-dependent and independent responses by direct action on B cells involving anti-apoptotic Bcl-2- and germinal center-promoting effects. AB - We recently developed a novel immunomodulating gene fusion protein, CTA1-DD, that combines the ADP-ribosylating ability of cholera toxin (CT) with a dimer of an Ig binding fragment, D, of Staphylococcus aureus protein A. The CTA1-DD adjuvant was found to be nontoxic and greatly augmented T cell-dependent responses to soluble protein Ags after systemic as well as mucosal immunizations. Here we show that CTA1-DD does not appear to form immune complexes or bind to soluble Ig following injections, but, rather, it binds directly to B cells of all isotypes, including naive IgD+ cells. No binding was observed to macrophages or dendritic cells. Immunizations in FcepsilonR (common FcRgamma-chain)- and FcgammaRII-deficient mice demonstrated that CTA1-DD exerted unaltered enhancing effects, indicating that FcgammaR-expressing cells are not required for the adjuvant function. Whereas CT failed to augment Ab responses to high m.w. dextran B512 in athymic mice, CTA1-DD was highly efficient, demonstrating that T cell-independent responses were also enhanced by this adjuvant. In normal mice both CT and CTA1 DD, but not the enzymatically inactive CTA1-R7K-DD mutant, were efficient enhancers of T cell-dependent as well as T cell-independent responses, and both promoted germinal center formation following immunizations. Although CT augmented apoptosis in Ag receptor-activated B cells, CTA1-DD strongly counteracted apoptosis by inducing Bcl-2 in a dose-dependent manner, a mechanism that was independent of the CD19 coreceptor. However, in the presence of CD40 stimulation, apoptosis was low and unaffected by CT, suggesting that the adjuvant effect of CT is dependent on the presence of activated CD40 ligand-expressing T cells. PMID- 10843682 TI - Vitamin D3 up-regulated protein 1 mediates oxidative stress via suppressing the thioredoxin function. AB - As a result of identifying the regulatory proteins of thioredoxin (TRX), a murine homologue for human vitamin D3 up-regulated protein 1 (VDUP1) was identified from a yeast two-hybrid screen. Cotransfection into 293 cells and precipitation assays confirmed that mouse VDUP1 (mVDUP1) bound to TRX, but it failed to bind to a Cys32 and Cys35 mutant TRX, suggesting the redox-active site is critical for binding. mVDUP1 was ubiquitously expressed in various tissues and located in the cytoplasm. Biochemical analysis showed that mVDUP1 inhibited the insulin-reducing activity of TRX. When cells were treated with various stress stimuli such as H2O2 and heat shock, mVDUP1 was significantly induced. TRX is known to interact with other proteins such as proliferation-associated gene and apoptosis signal regulating kinase 1. Coexpression of mVDUP1 interfered with the interaction between TRX and proliferation-associated gene or TRX and ASK-1, suggesting its roles in cell proliferation and oxidative stress. To investigate the roles of mVDUP1 in oxidative stress, mVDUP1 was overexpressed in NIH 3T3 cells. When cells were exposed to stress, cell proliferation was declined with elevated apoptotic cell death compared with control cells. In addition, c-Jun N-terminal kinase activation and IL-6 expression were elevated. Taken together, these results demonstrate that mVDUP1 functions as an oxidative stress mediator by inhibiting TRX activity. PMID- 10843683 TI - Correlation of T cell independence of antibody responses with antigen dose reaching secondary lymphoid organs: implications for splenectomized patients and vaccine design. AB - Many natural viral and bacterial pathogens activate B cells independently of Th cells (TI Ags). This study analyzed the characteristics of the activation of B cells after immunization with various forms of viral Ags using different immunization routes and found a decreasing dependence on T help with increasing amounts of Ag recruited to the spleen. Repetitive antigenic structure facilitated TI B cell responses if Ag was present in lymphoid organs. These results suggest that 1) Ag dose and localization in secondary lymphoid organs are the key for B cell activation in the absence of T help; 2) early TI Ab responses are crucial to protect against systemically spreading acute cytopathic infectious agents; and 3) there may be new rationales for improved vaccine design. PMID- 10843685 TI - Genetic immunization with lung-targeting macroaggregated polyethyleneimine albumin conjugates elicits combined systemic and mucosal immune responses. AB - Genetic immunization is a novel form of vaccination in which transgenes are delivered into the host to produce the foreign protein within host cells. Although systemic immune responses have been relatively easy to induce by genetic immunization, the induction of regional and mucosal immunity has often been more challenging. To address the problem of eliciting mucosal immunity in the lung, we utilized macroaggregated albumin to target plasmid DNA to the lung. Macroaggregated albumin is trapped in the lung after i. v. injection, and it is routinely used in radiolabeled form as an imaging modality to evaluate pulmonary blood flow. To couple DNA to this targeting agent, polyethyleneimine (a polycation that binds DNA and enhances transfection) was conjugated to serum albumin, and the conjugate was aggregated by heating to produce particles of 25 100 microm. The resulting particles bound plasmid DNA avidly, and when injected i.v. in mice, the particles distributed in the peripheral lung tissue in the alveolar interstitium. Particle-bound luciferase plasmid transfected a variety of cell lines in vitro, and after i.v. injection, gene expression was detected exclusively in the lung. Using human growth hormone as the encoded foreign Ag for immunization, i.v. injection of the particle-bound plasmid elicited both pulmonary mucosal and systemic immune responses, whereas naked DNA injected either i.v. or i.m. elicited only systemic responses. Thus, particle-bound plasmid DNA may have utility for genetic immunization by intravascular delivery to the lung and potentially to other organs and tissues. PMID- 10843684 TI - Mice with a selective deletion of the CC chemokine receptors 5 or 2 are protected from dextran sodium sulfate-mediated colitis: lack of CC chemokine receptor 5 expression results in a NK1.1+ lymphocyte-associated Th2-type immune response in the intestine. AB - The chemokine receptors CCR2 and CCR5 and their respective ligands regulate leukocyte chemotaxis and activation. To determine the role of these chemokine receptors in the regulation of the intestinal immune response, we induced colitis in CCR2- and CCR5-deficient mice by continuous oral administration of dextran sodium sulfate (DSS). Both CCR2- and CCR5-deficient mice were susceptible to DSS induced intestinal inflammation. The lack of CCR2 or CCR5 did not reduce the DSS induced migration of macrophages into the colonic lamina propria. However, both CCR5-deficient mice and, to a lesser degree, CCR2-deficient mice were protected from DSS-induced intestinal adhesions and mucosal ulcerations. CCR5-deficient mice were characterized by a greater relative infiltration of CD4+ and NK1.1+ lymphocyte in the colonic lamina propria when compared to wild-type and CCR2 deficient mice. In CCR5-deficient mice, mucosal mRNA expression of IL-4, IL-5, and IL-10 was increased, whereas that of IFN-gamma was decreased, corresponding to a Th2 pattern of T cell activation. In CCR2-deficient mice, the infiltration of Th2-type T cells in the lamina propria was absent, but increased levels of IL 10 and decreased levels of IFN-gamma may have down regulated mucosal inflammation. Our data indicate that CCR5 may be critical for the promotion of intestinal Th1-type immune responses in mice. PMID- 10843686 TI - The murine chemokine CXCL11 (IFN-inducible T cell alpha chemoattractant) is an IFN-gamma- and lipopolysaccharide-inducible glucocorticoid-attenuated response gene expressed in lung and other tissues during endotoxemia. AB - A new murine chemokine was identified in a search for glucocorticoid-attenuated response genes induced in the lung during endotoxemia. The first 73 residues of the predicted mature peptide are 71% identical and 93% similar to human CXCL11/IFN-inducible T cell alpha chemoattractant (I-TAC) (alias beta-R1, H174, IFN-inducible protein 9 (IP-9), and SCYB9B). The murine chemokine has six additional residues at the carboxyl terminus not present in human I-TAC. Identification of this cDNA as murine CXCL11/I-TAC is supported by phylogenetic analysis and by radiation hybrid mapping of murine I-TAC (gene symbol Scyb11) to mouse chromosome 5 close to the genes for monokine induced by IFN-gamma (MIG) and IP10. Murine I-TAC mRNA is induced in RAW 264.7 macrophages by IFN-gamma or LPS and is weakly induced by IFN-alphabeta. IFN-gamma induction of murine I-TAC is markedly enhanced by costimulation with LPS or IL-1beta in RAW cells and by TNF alpha in both RAW cells and Swiss 3T3 fibroblasts. Murine I-TAC is induced in multiple tissues during endoxemia, with strongest expression in lung, heart, small intestine, and kidney, a pattern of tissue expression different from those of MIG and IP10. Peak expression of I-TAC message is delayed compared with IP10, both in lung after i.v. LPS and in RAW 264.7 cells treated with LPS or with IFN gamma. Pretreatment with dexamethasone strongly attenuates both IFN-gamma-induced I-TAC expression in RAW cells and endotoxemia-induced I-TAC expression in lung and small intestine. The structural and regulatory similarities of murine and human I-TAC suggest that mouse models will be useful for investigating the role of this chemokine in human biology and disease. PMID- 10843687 TI - Functional analysis of the murine TCR beta-chain gene enhancer. AB - The TCR beta-chain gene enhancer activates transcription and V(D)J recombination in immature thymocytes. In this paper we present a systematic analysis of the elements that contribute to the activity of the murine TCR beta enhancer in mature and immature T cell lines. We identified a region containing the beta E4, beta E5, and beta E6 motifs as the essential core of the TCR beta enhancer in pro T cells. In mature cells, the core enhancer had low activity and required, in addition, either 5' or 3' flanking sequences whose functions may be partially overlapping. Mutation of any of the six protein binding sites located within the beta E4-beta E6 elements essentially abolished enhancer activity, indicating that this core enhancer contained no redundant elements. The beta E4 and beta E6 elements contain binding sites for ETS-domain proteins and the core binding factor. The beta E5 element bound two proteins that could be resolved chromatographically and that were both essential for enhancer activity. PMID- 10843688 TI - Primary structure and functional characterization of a soluble, alternatively spliced form of B7-1. AB - Recent studies have suggested that soluble forms of B7-1 and B7-2 may exist, but transcripts that code for these molecules have not been previously described. In this study, we report the cloning and characterization of an alternatively spliced soluble form of porcine B7-1 (sB7-1) that lacks exons coding for both the transmembrane and cytoplasmic domains. Northern blot analysis of RNA from alveolar macrophages revealed an approximate 3:1 ratio of the transmembrane form of B7-1 mRNA relative to sB7-1 mRNA. Porcine B7-1 was present on the surface of both B and T cells following stimulation with PMA/ionomycin. A histidine-tagged form of porcine sB7-1 (sB7-1-His) interacted with both CD28 and CTLA-4, and effectively blocked IL-2 production from human responder cells stimulated with PHA and either porcine or human stimulator cells. In addition, sB7-1-His inhibited human T cell proliferation in response to porcine or human peripheral blood leukocytes. This study is the first report of an alternatively spliced form of B7 that codes for a soluble protein. Furthermore, these data demonstrate that porcine B7-1 interacts with the human receptors CD28 and CTLA-4, suggesting a potential role for this molecule in pig to human xenotransplantation. Possible physiological functions for the soluble form of B7-1 are discussed. PMID- 10843689 TI - Regulation of TNF expression by multiple mitogen-activated protein kinase pathways. AB - Stimulating macrophages with bacterial endotoxin (LPS) activates numerous intracellular signaling pathways that lead to the production of TNF. In this study, we show that four mitogen-activated protein (MAP) kinase pathways are activated in LPS-stimulated macrophages: the extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase/stress-activated protein kinase, p38, and Big MAP kinase (BMK)/ERK5 pathways. Although specific activation of a single MAP kinase pathway produces only a modest effect on TNF promoter activation, activation of each MAP kinase pathway is important for full induction of the TNF gene. Interestingly, a dramatic induction of TNF promoter-driven gene expression was observed when all of the four MAP kinase pathways were activated simultaneously, suggesting a cooperative effect among these kinases. Unexpectedly, cis elements known to be targeted by MAP kinases do not play a major role in multiple MAP kinase-induced TNF gene expression. Rather, a 40-bp sequence harboring the TATA box, is responsible for the gene up-regulation induced by MAP kinases. The proximity of the MAP kinase-responsive element to the transcriptional initiation site suggested that MAP kinases regulate the transcriptional initiation complex. Utilizing alpha-amanitin-resistant RNA polymerase II mutants with or without a C-terminal domain (CTD) deletion, we found that deleting the CTD to 31 tandem repeats (Delta31) led to >90% reduction in MAP kinase-mediated TNF production. Thus, our data demonstrate coordination of multiple MAP kinase pathways in TNF production and suggest that the CTD of RNA polymerase II is required to execute MAP kinase signaling in TNF expression. PMID- 10843690 TI - Secretory phospholipase A2-potentiated inducible nitric oxide synthase expression by macrophages requires NF-kappa B activation. AB - The effect of secretory group II phospholipase A2 (sPLA2) on the expression of the inducible NO synthase (iNOS) and the production of NO by macrophages was investigated. sPLA2 by itself barely stimulated nitrite production and iNOS expression in Raw264.7 cells. However, in combination with LPS, the effects were synergistic. This potentiation was shown for sPLA2 enzymes from sPLA2-transfected stable cells or for purified sPLA2 from human synovial fluid. The effect of PLA2 on iNOS induction appears to be specific for the secretory type of PLA2. LPS stimulated activation of iNOS was inhibited by the well-known selective inhibitors of sPLA2 such as 12-epi-scalaradial and p-bromophenacyl bromide. In contrast, the cytosolic PLA2-specific inhibitors methyl arachidonyl fluorophosphate and arachidonyltrifluoromethyl ketone did not affect LPS-induced nitrite production and iNOS expression. Moreover, when we transfected cDNA encoding type II sPLA2, we observed that the sPLA2-transfected cells produced two times more nitrites than the empty vector or cytosolic PLA2-transfected cells. The sPLA2-potentiated iNOS expression was associated with the activation of NF kappa B. We found that the NF-kappa B inhibitor pyrrolidinedithiocarbamate prevented nitrite production, iNOS induction, and mRNA accumulation by sPLA2 plus LPS in Raw264.7 cells. Furthermore, EMSA analysis of the activation of the NF kappa B involved in iNOS induction demonstrated that pyrrolidinedithiocarbamate prevented the NF-kappa B binding by sPLA2 plus LPS. Our findings indicated that sPLA2, in the presence of LPS, is a potent activator of macrophages. It stimulates iNOS expression and nitrite production by a mechanism that requires the activation of NF-kappa B. PMID- 10843691 TI - Regulation of encephalitogenic T cells with recombinant TCR ligands. AB - We have previously described recombinant MHC class II beta1 and alpha1 domains loaded with free antigenic peptides with potent inhibitory activity on encephalitogenic T cells. We have now produced single-chain constructs in which the peptide Ag is genetically encoded within the same exon as the linked beta1 and alpha1 domains, overcoming the problem of displacement of peptide Ag from the peptide binding cleft. We here describe clinical effects of recombinant TCR ligands (RTLs) comprised of the rat RT1.B beta1alpha1 domains covalently linked to the 72-89 peptide of guinea pig myelin basic protein (RTL-201), to the corresponding 72-89 peptide from rat myelin basic protein (RTL-200), or to cardiac myosin peptide CM-2 (RTL-203). Only RTL-201 possessed the ability to prevent and treat active or passive experimental autoimmune encephalomyelitis. Amelioration of experimental autoimmune encephalomyelitis was associated with a selective inhibition of proliferation response and cytokine production by Ag stimulated lymph node T cells and a drastic reduction in the number of encephalitogenic and recruited inflammatory cells infiltrating the CNS. The exquisitely selective inhibition could be observed between molecules that differ by a single methyl group (the single amino acid residue difference between RTL 200 (threonine) and RTL-201 (serine) at position 80 of the myelin basic protein peptide). These novel RTLs provide a platform for developing potent and selective human diagnostic and therapeutic agents for treatment of autoimmune disease. PMID- 10843692 TI - Functional analysis of the OCA-B promoter. AB - OCA-B was identified as a B cell-specific coactivator that functions with either Oct-1 or Oct-2 to mediate efficient cell type-specific transcription via the octamer site (ATGCAAAT) both in vivo and in vitro. Mice lacking OCA-B exhibit normal Ag-independent B cell maturation. In contrast, Ag-dependent functions, including production of secondary Ig isotypes and germinal center formation, are greatly affected. To better understand OCA-B expression and, ultimately, the defects observed in the OCA-B knockout mice, we have cloned the OCA-B promoter and examined its function in both transformed and primary B cells. We show here that the OCA-B promoter is developmentally regulated, with activity increasing throughout B cell differentiation. Through physical and functional assays, we have found an activating transcription factor/cAMP response element binding protein binding site (or cAMP response element) that is crucial for OCA-B promoter activity. Furthermore, we demonstrate that IL-4 and anti-CD40 induce both the OCA-B promoter and octamer-dependent promoters, thus implicating OCA-B in B cell signaling events in the nucleus. PMID- 10843693 TI - Regulation of germline promoters by the two human Ig heavy chain 3' alpha enhancers. AB - The human IgH 3' enhancers, located downstream of each of the two Calpha genes, modulate germline (GL) transcription of the IgH genes by influencing the activity of promoter-enhancer complexes upstream of the switch and intervening (I) regions. The regulation of GL alpha1 and alpha2 promoters by different human 3' enhancer fragments was investigated in cell lines representing various developmental stages. Both alpha1HS1,2 and alpha2HS1,2 fragments show equally strong enhancer activity on the GL alpha1 and alpha2 promoters in both orientations when transiently transfected into a number of mature B cell line (DG75, CL-01, and HS Sultan). However, there is no activity in a human pre-B cell line (NALM-6) nor a human T cell line (Jurkat). HS3 shows no enhancer activity by itself in any of the cell lines, whereas a modest effect is noted using HS4 in the three mature B cell lines. However, the combination of the alpha2HS3-HS1,2 HS4 fragments, which together form a potential locus control region, displays a markedly stronger enhancer activity than the individual fragments with a differential effect on the alpha1 and alpha2 promoters as compared with the gamma3 promoter. Our results suggest that the human GL alpha promoter may be regulated by two independent pathways. One pathway is induced by TGF-beta1 which directs IgA isotype switch through activation of the GL alpha promoter and no TGF beta1-responsive elements are present in the different 3' enhancer fragments. The other route is through the human 3' enhancer regions that cis-up-regulate the GL alpha promoter activity in mature B cells. PMID- 10843694 TI - Evidence that terminal deoxynucleotidyltransferase expression plays a role in Ig heavy chain gene segment utilization. AB - TdT is a nuclear enzyme that catalyzes the addition of random nucleotides at Ig and TCR V(D)J junctions. In this paper we analyze human IgH rearrangements generated from transgenic minilocus mice in the presence or absence of TdT. In the absence of TdT, the pseudo-VH gene segment present in the minilocus is rearranged dramatically more frequently. Additionally, JH6 gene segment utilization is increased as well as the number of rearrangements involving only VH and JH gene segments. Thus, the recombination of IgH gene segments that are flanked by 23-nt spacer recombination signal sequences may be influenced by TdT expression. Extensive analysis indicates that these changes are independent of antigenic selection and cannot be explained by homology-mediated recombination. Thus, the role played by TdT may be more extensive than previously thought. PMID- 10843695 TI - The structure and stability of an HLA-A*0201/octameric tax peptide complex with an empty conserved peptide-N-terminal binding site. AB - The crystal structure of the human class I MHC molecule HLA-A2 complexed with of an octameric peptide, Tax8 (LFGYPVYV), from human T cell lymphotrophic virus-1 (HTLV-1) has been determined. This structure is compared with a newly refined, higher resolution (1.8 A) structure of HLA-A2 complexed with the nonameric Tax9 peptide (LLFGYPVYV) with one more N-terminal residue. Despite the absence of a peptide residue (P1) bound in the conserved N-terminal peptide-binding pocket of the Tax8/HLA-A2 complex, the structures of the two complexes are essentially identical. Water molecules in the Tax8 complex replace the terminal amino group of the Tax9 peptide and mediate a network of hydrogen bonds among the secondary structural elements at that end of the peptide-binding groove. Thermal denaturation measurements indicate that the Tax8 complex is much less stable, DeltaTm = 16 degrees C, than the Tax9 complex, but both can sensitize target cells for lysis by some Tax-specific CTL from HTLV-1 infected individuals. The absence of a P1 peptide residue is thus not enough to prevent formation of a "closed conformation" of the peptide-binding site. TCR affinity measurements and cytotoxic T cell assays indicate that the Tax8/HLA-A2 complex does not functionally cross-react with the A6-TCR-bearing T cell clone specific for Tax9/HLA-A2 complexes. PMID- 10843696 TI - IL-10 and the dangers of immune polarization: excessive type 1 and type 2 cytokine responses induce distinct forms of lethal immunopathology in murine schistosomiasis. AB - To dissect the controversial roles of type 1 and type 2 cytokines to the pathogenesis of schistosomiasis, we generated IL-10/IL-4- and IL-10/IL-12 deficient mice that develop highly polarized type 1 and type 2 cytokine responses, respectively. Interestingly, the Th1-polarized IL-10/IL-4-deficient mice rapidly lost weight at the onset of egg-laying and displayed 100% mortality by wk 9 postinfection. This acute mortality was linked to overexpression of the proinflammatory mediators IFN-gamma, TNF-alpha, and inducible NO and the formation of nonfibrotic granulomas. Elevated serum aspartate transaminase levels confirmed that mortality was in part attributable to acute hepatotoxicity. In contrast, the Th2-polarized IL-10/IL-12-deficient mice developed a progressive wasting disease that correlated with increased hepatic fibrosis, formation of large eosinophil-rich granulomas, a 10-fold increase in IL-4 and IL-13, and significant mortality during the chronic stages of infection. Surprisingly, IL-10 deficient mice displayed pathological features that were characteristic of both extremes, while wild-type mice developed relatively successful long term chronic infections. These data demonstrate that IL-10 significantly suppresses type 1 and type 2 cytokine development in IL-4- and IL-12-deficient mice, respectively, thereby impeding the development of severe egg-induced pathology in the single cytokine-deficient animals. Together, these findings reveal the central regulatory role of IL-10 in the pathogenesis of schistosomiasis and illustrate that excessive type 1 and type 2 cytokine responses trigger distinct, but equally detrimental, forms of pathology following infection. PMID- 10843697 TI - Infection of B cell-deficient mice with CDC 1551, a clinical isolate of Mycobacterium tuberculosis: delay in dissemination and development of lung pathology. AB - Long-term survival of mice infected with Mycobacterium tuberculosis is dependent upon IFN-gamma and T cells, but events in early phases of the immune response are not well understood. In this study, we describe a role for B cells during early immune responses to infection with a clinical isolate of M. tuberculosis (CDC 1551). Following a low-dose infection with M. tuberculosis CDC 1551, similar numbers of bacteria were detected in the lungs of both B cell knockout (IgH 6-, BKO) and C57BL/6J (wild-type) mice. However, despite comparable bacterial loads in the lungs, less severe pulmonary granuloma formation and delayed dissemination of bacteria from lungs to peripheral organs were observed in BKO mice. BKO mice reconstituted with naive B cells, but not those given M. tuberculosis-specific Abs, before infection developed pulmonary granulomas and dissemination patterns similar to wild-type animals. Further analysis of lung cell populations revealed greater numbers of lymphocytes, especially CD8+ T cells, macrophages, and neutrophils in wild-type and reconstituted mice than in BKO mice. Thus, less severe lesion formation and delayed dissemination of bacteria found in BKO mice were dependent on B cells, not Abs, and were associated with altered cellular infiltrate to the lungs. These observations demonstrate an important, previously unappreciated, role for B cells during early immune responses to M. tuberculosis infections. PMID- 10843698 TI - Modulation of HLA-G antigens expression by human cytomegalovirus: specific induction in activated macrophages harboring human cytomegalovirus infection. AB - After infection, human CMV (HCMV) establishes a latent and persistent infection in immature myeloid progenitors and peripheral blood monocytes. Completion of the HCMV life cycle is possible upon maturation of monocytes to tissue macrophages and under permissive circumstances, e.g., immunosuppression. We investigated the hypothesis that HLA-G molecules could be induced during HCMV reactivation in activated macrophages to favor virus dissemination. In this study, we provide evidence that HLA-G Ags are produced during viral reactivation in macrophages generated after allogeneic stimulation of HCMV latently infected monocytes. While HLA-G surface expression is up-regulated, classical MHC-I molecules are partially down-regulated by HCMV. In vivo, bronchoalveolar macrophages collected from patients suffering from acute HCMV pneumonitis also express HLA-G molecules. The direct correlation between HLA-G Ag induction and HCMV infection was confirmed in U-373 MG astrocytoma cells. Soluble HLA-G expression is stimulated upon HCMV infection, and this modulation depends on the cooperative action of the two immediate-early-1 pp72 and immediate-early-2 pp86 products. Because HLA-G transcription is active in macrophages and U-373 MG astrocytoma cells, it is likely that the modulation of HLA-G protein expression during HCMV replication occurs at a post-transcriptional level. Our data suggest that induction of HLA-G molecules could be an additional mechanism that helps HCMV to subvert host defenses. PMID- 10843699 TI - Plasmid DNA encoding IFN-alpha 1 antagonizes herpes simplex virus type 1 ocular infection through CD4+ and CD8+ T lymphocytes. AB - The present study was undertaken to further characterize the anti-viral efficacy of a plasmid DNA encoding IFN-alpha1 against ocular herpes simplex virus type 1 (HSV-1) infection. In mice ocularly treated with plasmid DNA encoding IFN-alpha 1, the efficacy of the transgene was inversely proportional to the amount of virus used to infect the mice. Ocular treatment of mice with the IFN-alpha 1 transgene was the only mucosal route tested that showed efficacy against ocular HSV-1 infection compared with vaginal or intranasal delivery. Mice treated with the plasmid DNA encoding IFN-alpha 1 showed a significant reduction in viral Ag expression in the eyes and trigeminal ganglion that correlated with a reduction in immune cell infiltration into the cornea and iris on days 3 and 6 postinfection, as evidenced by immunohistochemical staining. Depleting mice of either CD4+ or CD8+ T lymphocytes completely blocked the resistance to herpes simplex virus type 1-induced mortality in mice treated with the IFN-alpha 1 transgene. In the absence of infection, the application of naked DNA encoding IFN alpha 1 significantly increased the levels of IL-6- and IFN-gamma-inducible protein 10 transcript expression in the corneas 24 h post-treatment. Expression of the plasmid construct following topical application in the eye included the rectus muscles proximal to the cornea as well as the spleen. Collectively, the protective efficacy of the IFN-alpha 1 transgene against ocular HSV-1 infection is dependent upon the local or distal participation of CD4+ and CD8+ T lymphocytes early in the course of the infection, suggesting an indirect effect of the transgene against HSV-1-induced mortality. PMID- 10843700 TI - Adaptive immunity and enhanced CD8+ T cell response to Listeria monocytogenes in the absence of perforin and IFN-gamma. AB - Single Ag-specific CD8+ T cells from IFN-gamma-deficient (GKO) or perforin deficient (PKO) mice provide substantial immunity against murine infection with Listeria monocytogenes. To address the potential for redundancy between perforin and IFN-gamma as CD8+ T cell effector mechanisms, we generated perforin/IFN-gamma (PKO/GKO) double-deficient mice. PKO/GKO-derived CD8+ T cells specific for the immunodominant listeriolysin O (LLO91-99) epitope provide immunity to LM infection similar to that provided by Ag-matched wild-type (WT) CD8+ T cells in the liver but reduced in the spleen. Strikingly, polyclonal CD8+ T cells from immunized PKO/GKO mice were approximately 100-fold more potent in reducing bacterial numbers than the same number of polyclonal CD8+ T cells from immunized WT mice. This result is probably quantitative, because the frequency of the CD8+ T cell response against the immunodominant LLO91-99 epitope is >4.5-fold higher in PKO/GKO mice than WT mice at 7 days after identical immunizations. Moreover, PKO/GKO mice can be immunized by a single infection with attenuated Listeria to resist >80,000-fold higher challenges with virulent organisms than naive PKO/GKO mice. These data demonstrate that neither perforin nor IFN-gamma is required for the development or expression of adaptive immunity to LM. In addition, the results suggest the potential for perforin and IFN-gamma to regulate the magnitude of the CD8+ T cell response to infection. PMID- 10843701 TI - A filarial nematode-secreted product signals dendritic cells to acquire a phenotype that drives development of Th2 cells. AB - Although exogeneous "danger" signals such as LPS can activate APC to produce a Th1 response, the nature of events initiating a Th2 response is controversial. We now show that pathogen-derived products have the capacity to induce bone marrow derived dendritic cell cultures to acquire a phenotype that promotes the differentiation of naive CD4+ T cells toward either a Th1 or Th2 phenotype. Thus, LPS-matured dendritic cells (DC1) promote a Th1 response (increased generation of IFN-gamma and reduced production of IL-4) by Ag-stimulated CD4+ T cells from the DO.11.10 transgenic mouse expressing a TCR specific for an OVA peptide (OVA323 339). In contrast, a phosphorylcholine-containing glycoprotein, ES-62, secreted by the filarial nematode, Acanthocheilonema viteae, which generates a Th2 Ab response in vivo, is found to induce the maturation of dendritic cells (DC2) with the capacity to induce Th2 responses (increased IL-4 and decreased IFN-gamma). In addition, we show that the switch to either Th1 or Th2 responses is not effected by differential regulation through CD80 or CD86 and that a Th2 response is achieved in the presence of IL-12. PMID- 10843702 TI - Delta 9-tetrahydrocannabinol treatment suppresses immunity and early IFN-gamma, IL-12, and IL-12 receptor beta 2 responses to Legionella pneumophila infection. AB - The marijuana cannabinoid, delta 9-tetrahydrocannabinol (THC), suppresses immunity to Legionella pneumophila and development of Th1 activity and cell mediated immunity. In the current study, THC effects on cytokines regulating the development of Th1 cells were examined. BALB/c mice showed significant increases in serum IL-12 and IFN-gamma within hours of infection; however, the levels of these Th1-promoting cytokines as well as resistance to a challenge infection were suppressed by THC (8 mg/kg) injected 18 h before priming. The Th2-promoting cytokine, IL-4, was increased within hours of a Legionella infection and was further increased by THC treatment. These results suggested that THC injection suppressed the cytokine environment promoting Th1 immunity. In additional experiments, THC pretreatment and infection of IL-4 knockout mice showed that serum IL-12 and IFN-gamma were suppressed equally in both knockout and normal mice. This suggested that the drug-induced increase in IL-4 was not responsible for the decreases in serum IL-12 and IFN-gamma. However, THC treatment was shown to suppress the expression of IL-12 receptor beta 2 mRNA, indicating that, in addition to suppression of IL-12, THC injection suppressed the expression of IL 12 receptors. Finally, the role of cannabinoid receptors in Th1-promoting cytokine suppression was examined, and results with receptor antagonists showed that both cannabinoid receptors 1 and 2 were involved. It is suggested that suppression of Th1 immunity to Legionella is not due to an increase in IL-4 production but to a decrease in IFN-gamma and IL-12. Furthermore, both types of cannabinoid receptors are involved. PMID- 10843703 TI - EBV suppresses prostaglandin E2 biosynthesis in human monocytes. AB - It is well known that EBV has developed strategies to evade immune surveillance. Previously, EBV was shown to bind specifically to monocytes and regulate expression of proinflammatory mediators such as IL-1, IL-6, TNF-alpha, and leukotrienes. EBV was also found to affect phagocytosis of monocytes. In this study, we show that in addition to these effects, EBV suppresses the biosynthesis of PGE2, a pleiotropic immunomodulatory molecule that is synthesized by the dioxygenation of arachidonic acid via the cyclooxygenase (COX) pathway. This down regulation of PGE2 formation involved the inhibition of the inducible COX-2 isoform expression both at the transcriptional and translational levels, whereas expression of the constitutive COX-1 isoform was unaltered. Furthermore, exposure of monocytes to EBV was found to impact on the NF-kappaB activation pathway, which plays an essential role in the induction of COX-2 in monocytes. The inhibition of PGE2 biosynthesis was relieved when the experiments were conducted in presence of phosphonoacetic acid, an inhibitor of herpesviruses DNA polymerase, indicating that viral replication and/or neosynthesized viral proteins were involved in this process. Thus, inhibition of PGE2 biosynthesis in monocytes may represent an additional mechanism underlying EBV pathogenicity. PMID- 10843704 TI - Simian immunodeficiency virus evades a dominant epitope-specific cytotoxic T lymphocyte response through a mutation resulting in the accelerated dissociation of viral peptide and MHC class I. AB - The ability of an AIDS virus to escape from immune containment by selective mutation away from recognition by CTL was explored in simian immunodeficiency virus of macaques (SIVmac)-infected rhesus monkeys. CTL recognition of a previously defined common viral mutation in an immunodominant SIVmac Gag epitope was evaluated. CTL were assessed for their ability to recognize a SIVmac Gag protein with a single residue 2 (T --> A) replacement in the minimal epitope peptide bound by the MHC class I molecule Mamu-A*01. SIVmac Gag-specific CTL lysed Mamu-A*01+ target cells infected with recombinant vaccinia virus expressing the wild-type but not the mutant Gag protein. In addition, CTL recognized the mutant epitope peptide less efficiently than the wild-type virus peptide. In studies to determine the mechanism by which the mutant virus evaded CTL recognition, this peptide was shown to bind Mamu-A*01 in a manner that was indistinguishable from the wild-type peptide. However, experiments in which an increasing duration of delay was introduced between peptide sensitization of target cells and the assessment of these cells as targets in killing assays suggest that the mutant peptide with a T --> A replacement had a higher off-rate from Mamu-A*01 than the wild-type peptide did. Therefore, these findings suggest that AIDS viruses can evade virus-specific CTL responses through the accelerated dissociation of mutant peptide from MHC class I. PMID- 10843705 TI - NK cells cause liver injury and facilitate the induction of T cell-mediated immunity to a viral liver infection. AB - NK cells are a relatively rare cell population in peripheral lymphoid organs but are abundant in the liver, raising questions as to their function in immune responses to infections of this organ. To investigate this, cell-mediated immunity to viral liver infection induced by a type 5, replication-defective, adenovirus was examined. It is shown that NK cells in the absence of T cells cause hepatocyte apoptosis in virus-infected livers associated with an increase in liver enzymes in the serum. Concomitantly, NK cells induce production of IFN gamma, inhibitable by their elimination before infection. NK cells are shown to be necessary for optimal priming of virus-specific T cells, assessed by delayed type hypersensitivity response and CTL activity, consistent with their ability to secrete IFN-gamma. The conclusion is drawn that NK cells mediate two important functions in the liver: they induce cell death in the infected organ and concomitantly stimulate the induction of T cell-mediated immunity by release of IFN-gamma. PMID- 10843706 TI - Neutrophil-induced changes in the biomechanical properties of endothelial cells: roles of ICAM-1 and reactive oxygen species. AB - This study evaluated the changes in the biomechanical properties of endothelial cells (ECs) induced by neutrophil adhesion and the roles of ICAM-1 and reactive oxygen species (ROS) in modulating these changes. Neutrophil adherence to 24-h TNF-alpha-activated pulmonary microvascular ECs induced an increase in the apparent stiffness of ECs within 2 min, measured with magnetic twisting cytometry. An anti-ICAM-1 Ab blocked the EC stiffening response without inhibiting neutrophil adherence. Moreover, cross-linking ICAM-1 mimicked the stiffening response induced by neutrophils. The neutrophil-induced increase in the apparent stiffness of ECs was inhibited with 1% DMSO (a hydroxyl radical scavenger), allopurinol (a xanthine oxidase inhibitor), or deferoxamine (an iron chelator), suggesting that ROS may be involved in mediating the EC stiffening response. The cellular sources of ROS were determined by measuring the oxidation of dichlorofluorescein. Neutrophil adherence to TNF-alpha-activated ECs induced ROS production only in ECs, and not in neutrophils. This ROS production in ECs was completely prevented by the anti-ICAM-1 Ab and partially inhibited by allopurinol. These results suggest that ICAM-1-mediated signaling events during neutrophil adherence may activate xanthine oxidase, which in turn mediates the ROS production in ECs that leads to stiffening. ROS generated in ECs on neutrophil adherence appear to mediate cytoskeletal remodeling, which may modulate subsequent inflammatory responses. PMID- 10843707 TI - Combined effects of IL-12 and IL-18 on the induction of collagen-induced arthritis. AB - IL-18 expression has recently been detected in rheumatoid arthritis (RA) synovial membrane. We investigated the mechanisms by which IL-18-induced collagen-induced arthritis in DBA/1 mice primed intradermally with type II bovine collagen in IFA and boosted i.p. 21 days later with CII in saline. Mice were injected i.p. with rIL-12, rIL-18, or both (100 ng) during days -1 to 4 and again on days 20-24. Control mice received PBS. Mice treated with IL-12 or IL-18 alone developed significantly higher incidence and more severe disease compared with controls. These were elevated further by combination treatment with IL-12 and IL-18. The cytokine treatments led to markedly enhanced synovial hyperplasia, cellular infiltration, and cartilage erosion compared with controls. Cytokine-treated mice produced significantly more IFN-gamma, TNF-alpha, and IL-6 than the controls. Interestingly, IL-18-treated mice produced more TNF-alpha and IL-6, but less IFN gamma, compared with mice treated with IL-12. Furthermore, splenic macrophages from DBA/1 mice cultured in vitro with IL-18, but not IL-12, produced substantial amounts of TNF-alpha. Mice treated with IL-18 or IL-18 plus IL-12 produced markedly more IgG1 and IgG2a anti-collagen Ab compared with controls, whereas IL 12 treatment only led to an enhanced IgG2a response. Together these results demonstrate that IL-18 can promote collagen-induced inflammatory arthritis through mechanisms that may be distinct from those induced by IL-12. PMID- 10843709 TI - Resveratrol suppresses TNF-induced activation of nuclear transcription factors NF kappa B, activator protein-1, and apoptosis: potential role of reactive oxygen intermediates and lipid peroxidation. AB - Resveratrol (trans-3,4',5-trihydroxystilbene), a polyphenolic phytoalexin found in grapes, fruits, and root extracts of the weed Polygonum cuspidatum, exhibits anti-inflammatory, cell growth-modulatory, and anticarcinogenic effects. How this chemical produces these effects is not known, but it may work by suppressing NF kappaB, a nuclear transcription factor that regulates the expression of various genes involved in inflammation, cytoprotection, and carcinogenesis. In this study, we investigated the effect of resveratrol on NF-kappaB activation induced by various inflammatory agents. Resveratrol blocked TNF-induced activation of NF kappaB in a dose- and time-dependent manner. Resveratrol also suppressed TNF induced phosphorylation and nuclear translocation of the p65 subunit of NF kappaB, and NF-kappaB-dependent reporter gene transcription. Suppression of TNF induced NF-kappaB activation by resveratrol was not restricted to myeloid cells (U-937); it was also observed in lymphoid (Jurkat) and epithelial (HeLa and H4) cells. Resveratrol also blocked NF-kappaB activation induced by PMA, LPS, H2O2, okadaic acid, and ceramide. The suppression of NF-kappaB coincided with suppression of AP-1. Resveratrol also inhibited the TNF-induced activation of mitogen-activated protein kinase kinase and c-Jun N-terminal kinase and abrogated TNF-induced cytotoxicity and caspase activation. Both reactive oxygen intermediate generation and lipid peroxidation induced by TNF were suppressed by resveratrol. Resveratrol's anticarcinogenic, anti-inflammatory, and growth modulatory effects may thus be partially ascribed to the inhibition of activation of NF-kappaB and AP-1 and the associated kinases. PMID- 10843710 TI - Phagocytosis of nonapoptotic cells dying by caspase-independent mechanisms. AB - Caspase activation, exposure of phosphatidylserine (PS) on the outer surface of the plasma membrane, and rapid phagocytic removal of dying cells are key features of apoptosis. Nonapoptotic/necrotic modes of death occur independent of caspase activation, but the role of phagocytosis is largely unknown. To address this issue, we studied phagocytosis by human monocyte-derived macrophages (HMDM) and rat microglial cells. Target cells (Jurkat) were stimulated by several different methods that all caused caspase-independent death. First, we induced necrosis by combining toxins with ATP-depleting agents. Under these conditions, neither PS was exposed nor were such cells phagocytosed before their death. However, once the plasma membrane integrity was lost, the dead cells were rapidly and efficiently engulfed by HMDM. Next, we triggered Jurkat cell death with staurosporine in the presence of the pan-caspase inhibitor zVAD-fmk. Under these conditions, death occurred by delayed necrosis and without exposure of PS. Nevertheless, such lethally challenged cells were phagocytosed before the loss of membrane integrity. Finally, we triggered Ca2+ influx in Jurkat cells with an ionophore, or in neurons by glutamate receptor stimulation, respectively. In both models, PS was exposed on the cell surface. Ca2+-stressed cells were phagocytosed starting at 30 min after stimulation. Protein kinase C inhibitors prevented Ca2+ mediated PS exposure and phagocytosis. Essentially, similar phagocytosis data were obtained for all models with HMDM and microglia. We conclude that also cells dying nonapoptotically and independent of caspase activation may be recognized and removed before, or very quickly after, membrane lysis. PMID- 10843708 TI - Peroxisome proliferator-activated receptor-gamma activators inhibit IFN-gamma induced expression of the T cell-active CXC chemokines IP-10, Mig, and I-TAC in human endothelial cells. AB - Peroxisome proliferator-activated receptor-gamma (PPARgamma), a member of the nuclear hormone receptor superfamily originally shown to play an important role in adipocyte differentiation and glucose homeostasis, is now known to regulate inflammatory responses. Given the importance of endothelial cell (EC)-derived chemokines in regulating leukocyte function and trafficking, we studied the effects of PPARgamma ligands on the expression of chemokines induced in ECs by the Th1 cytokine IFN-gamma. Treatment of ECs with PPARgamma activators significantly inhibited IFN-gamma-induced mRNA and protein expression of the CXC chemokines IFN-inducible protein of 10 kDa (IP-10), monokine induced by IFN-gamma (Mig), and IFN-inducible T-cell alpha-chemoattractant (I-TAC), whereas expression of the CC chemokine monocyte chemoattractant protein-1 was not altered. PPARgamma activators decreased IFN-inducible protein of 10 kDa promoter activity and inhibited protein binding to the two NF-kappaB sites but not to the IFN stimulated response element ISRE site. Furthermore, PPARgamma ligands inhibited the release of chemotactic activity for CXC chemokine receptor 3 (CXCR3) transfected lymphocytes from IFN-gamma-stimulated ECs. These data suggest that anti-diabetic PPARgamma activators might attenuate the recruitment of activated T cells at sites of Th1-mediated inflammation. PMID- 10843711 TI - Differential mitogen-activated protein kinase stimulation by Fc gamma receptor IIa and Fc gamma receptor IIIb determines the activation phenotype of human neutrophils. AB - Fc gamma Rs mediate immune complex-induced tissue injury. The hypothesis that Fc gamma RIIa and Fc gamma RIIIb control neutrophil responses by activating mitogen activated protein kinases was examined. Homotypic and heterotypic cross-linking of Fc gamma RIIa and/or Fc gamma RIIIb resulted in a rapid, transient increase in ERK and p38 activity, with maximal stimulation between 1 and 3 min. Fc gamma RIIa and Fc gamma RIIIb stimulated distinct patterns of ERK and p38 activity, and heterotypic cross-linking failed to stimulate synergistic activation of either ERK or p38 activity. Both Fc gamma RIIa and Fc gamma RIIIb required activation of a nonreceptor tyrosine kinase and phosphatidylinositol 3-kinase for stimulation of ERK and p38. Inhibition of ERK activation with PD98059 enhanced H2O2 production stimulated by homotypic and heterotypic Fc gamma R cross-linking. Inhibition of p38 with SB203580 attenuated H2O2 production stimulated by Fc gamma RIIIb or heterotypic cross-linking, but had no effect on Fc gamma RIIa-stimulated H2O2 production. On the other hand, PD98059 inhibited actin polymerization stimulated by Fc gamma R cross-linking, while SB203580 had no effect. Inhibition of actin polymerization with cytochalasin D enhanced p38 activity stimulated by either Fc gamma RIIa or Fc gamma RIIIb, but cytochalasin D only enhanced H2O2 production stimulated by Fc gamma RIIIb. Our data indicate that Fc gamma RIIa and Fc gamma RIIIb independently activate ERK and p38. The two receptors demonstrate different efficacies for ERK and p38 activation, and they do not act cooperatively. ERK and p38 provide stimulatory and inhibitory signals for neutrophil responses to immune complexes. In addition, these data indicate that actin reorganization may play a role in mediating p38-dependent activation of respiratory burst upon stimulation of Fc gamma RIIIb in neutrophils. PMID- 10843712 TI - Release of calcium from inositol 1,4,5-trisphosphate receptor-regulated stores by HIV-1 Tat regulates TNF-alpha production in human macrophages. AB - HIV-1 protein Tat is neurotoxic and increases macrophage and microglia production of TNF-alpha, a cytopathic cytokine linked to the neuropathogenesis of HIV dementia. Others have shown that intracellular calcium regulates TNF-alpha production in macrophages, and we have shown that Tat releases calcium from inositol 1,4, 5-trisphosphate (IP3) receptor-regulated stores in neurons and astrocytes. Accordingly, we tested the hypothesis that Tat-induced TNF-alpha production was dependent on the release of intracellular calcium from IP3 regulated calcium stores in primary macrophages. We found that Tat transiently and dose-dependently increased levels of intracellular calcium and that this increase was blocked by xestospongin C, pertussis toxin, and by phospholipase C and type 1 protein kinase C inhibitors but not by protein kinase A or phospholipase A2 inhibitors. Xestospongin C, BAPTA-AM, U73122, and bisindolylmalemide significantly inhibited Tat-induced TNF-alpha production. These results demonstrate that in macrophages, Tat-induced release of calcium from IP3-sensitive intracellular stores and activation of nonconventional PKC isoforms play an important role in Tat-induced TNF-alpha production. PMID- 10843713 TI - Cyclic AMP blocks bacterial lipopolysaccharide-induced myosin light chain phosphorylation in endothelial cells through inhibition of Rho/Rho kinase signaling. AB - During Gram-negative sepsis bacterial LPS induces endothelial cell contraction, actin reorganization, and loss of endothelial integrity by an unknown signal mechanism. In this study, we provide evidence that LPS-stimulation of endothelial cells (HUVEC) decreases myosin light chain (MLC) phosphatase, resulting in an increase in MLC phosphorylation followed by cell contraction. All of these LPS effects could be blocked by the Rho-GTPase inhibitor C3 transferase from Clostridium botulinum or the Rho kinase inhibitor Y-27632. These data suggest that LPS induces MLC phosphorylation via Rho/Rho kinase-mediated inhibition of MLC phosphatase in HUVEC. Furthermore, we observed that cAMP-elevating drugs, known to exert a vasoprotective function, mimicked the effects of C3 transferase and Y-27632, i.e., inhibited LPS-induced MLC phosphatase inactivation and MLC phosphorylation. cAMP elevation did not inhibit myosin phosphorylation induced by constitutively active V14Rho or the MLC phosphatase inhibitor calyculin and did not induce phosphorylation of RhoA in HUVEC, indicating inhibition of an upstream regulator of Rho/Rho kinase. Taken together, Rho/Rho kinase appears to be a central target for inflammatory mediators causing endothelial cell contraction such as bacterial toxins, but also for vasoprotective molecules elevating intracellular cAMP. PMID- 10843714 TI - Lymphocyte migration through monolayers of endothelial cell lines involves VCAM-1 signaling via endothelial cell NADPH oxidase. AB - Lymphocytes migrate from the blood across endothelial cells to reach foreign substances sequestered in peripheral lymphoid organs and inflammatory sites. To study intracellular signaling in endothelial cells during lymphocyte migration, we used murine endothelial cell lines that promote lymphocyte migration and constitutively express VCAM-1. The maximum rate of resting splenic lymphocyte migration across monolayers of the endothelial cells occurred at 0-24 h. This migration was inhibited by anti-VCAM-1 or anti-alpha4 integrin, suggesting that VCAM-1 adhesion was required for migration. To determine whether signals within the endothelial cells were required for migration, irreversible inhibitors of signal transduction molecules were used to pretreat the endothelial cell lines. Inhibitors of NADPH oxidase activity (diphenyleneiodonium and apocynin) blocked migration >65% without affecting adhesion. Because NADPH oxidase catalyzes the production of reactive oxygen species (ROS), we examined whether ROS were required for migration. Scavengers of ROS inhibited migration without affecting adhesion. Furthermore, VCAM-1 ligand binding stimulated NADPH oxidase-dependent production of ROS by the endothelial cells lines and primary endothelial cell cultures. Finally, VCAM-1 ligand binding induced an apocynin-inhibitable actin restructuring in the endothelial cell lines at the location of the lymphocyte or anti-VCAM-1-coated bead, suggesting that an NADPH oxidase-dependent endothelial cell shape change was required for lymphocyte migration. In summary, VCAM-1 signaled the activation of endothelial cell NADPH oxidase, which was required for lymphocyte migration. This suggests that endothelial cells are not only a scaffold for lymphocyte adhesion, but play an active role in promoting lymphocyte migration. PMID- 10843715 TI - A new small molecule C5a receptor antagonist inhibits the reverse-passive Arthus reaction and endotoxic shock in rats. AB - C5a is implicated as a pathogenic factor in a wide range of immunoinflammatory diseases, including sepsis and immune complex disease. Agents that antagonize the effects of C5a could be useful in these diseases. We have developed some novel C5a antagonists and have determined the acute anti-inflammatory properties of a new small molecule C5a receptor antagonist against C5a- and LPS-induced neutrophil adhesion and cytokine expression, as well as against some hallmarks of the reverse Arthus reaction in rats. We found that a single i.v. dose (1 mg/kg) of this antagonist inhibited both C5a- and LPS-induced neutropenia and elevated levels of circulating TNF-alpha, as well as polymorphonuclear leukocyte migration, increased TNF-alpha levels and vascular leakage at the site of immune complex deposition. These results indicate potent anti-inflammatory activities of a new C5a receptor antagonist and provide more evidence for a key early role for C5a in sepsis and the reverse Arthus reaction. The results support a role for antagonists of C5a receptors in the therapeutic intervention of immunoinflammatory disease states such as sepsis and immune complex disease. PMID- 10843716 TI - Characterization of human gamma-tryptases, novel members of the chromosome 16p mast cell tryptase and prostasin gene families. AB - Previously, this laboratory identified clusters of alpha-, beta-, and mast cell protease-7-like tryptase genes on human chromosome 16p13.3. The present work characterizes adjacent genes encoding novel serine proteases, termed gamma tryptases, and generates a refined map of the multitryptase locus. Each gamma gene lies between an alpha1H Ca2+ channel gene (CACNA1H) and a betaII- or betaIII tryptase gene and is approximately 30 kb from polymorphic minisatellite MS205. The tryptase locus also contains at least four tryptase-like pseudogenes, including mastin, a gene expressed in dogs but not in humans. Genomic DNA blotting results suggest that gammaI- and gammaII-tryptases are alleles at the same site. betaII- and betaIII-tryptases appear to be alleles at a neighboring site, and alphaII- and betaI-tryptases appear to be alleles at a third site. gamma-Tryptases are transcribed in lung, intestine, and in several other tissues and in a mast cell line (HMC-1) that also expresses gamma-tryptase protein. Immunohistochemical analysis suggests that gamma-tryptase is expressed by airway mast cells. gamma-Tryptase catalytic domains are approximately 48% identical with those of known mast cell tryptases and possess mouse homologues. We predict that gamma-tryptases are glycosylated oligomers with tryptic substrate specificity and a distinct mode of activation. A feature not found in described tryptases is a C terminal hydrophobic domain, which may be a membrane anchor. Although the catalytic domains contain tryptase-like features, the hydrophobic segment and intron-exon organization are more closely related to another recently described protease, prostasin. In summary, this work describes gamma-tryptases, which are novel members of chromosome 16p tryptase/prostasin gene families. Their unique features suggest possibly novel functions. PMID- 10843718 TI - Recurrent respiratory syncytial virus infections in allergen-sensitized mice lead to persistent airway inflammation and hyperresponsiveness. AB - Respiratory syncytial virus (RSV) infection is considered a risk factor for bronchial asthma; however, the synergy between allergen sensitization and RSV infection in the development of pulmonary inflammation and asthma has been controversial. In this study the effects of primary and recurrent RSV infection on allergic asthma were examined in a group of control, RSV-infected, Dermatophagoides farinae (Df) allergen-sensitized, and Df allergen-sensitized plus RSV-infected BALB/c mice. Primary RSV infection in Df-sensitized mice transiently increases airway responsiveness, which is accompanied by increases in eosinophilic infiltration, the expression of ICAM-1, and macrophage inflammatory protein-1alpha (MIP-1alpha) in the lung tissue. A secondary RSV infection persistently enhances airway responsiveness in Df-sensitized mice, with a concomitant increase in MIP-1alpha and RSV Ag load in lung tissues. Bulk cultures of thoracic lymph node mononuclear cells demonstrate that acute RSV infection augments both Th1- and Th2-like cytokines, whereas secondary and tertiary infections shift the cytokine profile in favor of the Th2-like cytokine response in Df-sensitized mice. The elevated total serum IgE level in the Df-sensitized mice persists following only RSV reinfection. Thus, recurrent RSV infections in Df-sensitized mice augment the synthesis of Th2-like cytokines, total serum IgE Abs, and MIP-1alpha, which are responsible for persistent airway inflammation and hyperresponsiveness, both of which are characteristics of asthma. PMID- 10843717 TI - Prolonged elevation of IL-1 in Pseudomonas aeruginosa ocular infection regulates macrophage-inflammatory protein-2 production, polymorphonuclear neutrophil persistence, and corneal perforation. AB - The kinetics of IL-1 (alpha and beta) production after Pseudomonas aeruginosa corneal infection was examined in susceptible (cornea perforates) C57BL/6J (B6) and resistant (cornea heals) BALB/cByJ (BALB/c) mice. IL-1alpha and -1beta (mRNA and protein) were elevated in both mouse strains, and levels peaked at 1 day postinfection (p.i. ). Significantly greater amounts of IL-1 protein were detected in B6 vs BALB/c mice at 1 and 3 days p.i. At 5 days p.i., IL-1alpha and 1beta (mRNA and protein) remained elevated in B6, but began to decline in BALB/c mice. To test the significance of elevated IL-1 in B6 mice, a polyclonal neutralizing Ab against IL-1beta was used to treat infected B6 mice. A combination of subconjunctival and i.p. administration of IL-1beta polyclonal Ab significantly reduced corneal disease. The reduction in disease severity in infected B6 mice was accompanied by a reduction in corneal polymorphonuclear neutrophil number, bacterial load, and macrophage inflammatory protein-2 mRNA and protein levels. These data provide evidence that IL-1 is an important contributor to P. aeruginosa corneal infection. At least one mechanism by which prolonged and/or elevated IL-1 expression contributes to irreversible corneal tissue destruction appears to be by increasing macrophage inflammatory protein-2 production, resulting in a prolonged stimulation of polymorphonuclear neutrophil influx into cornea. In contrast, a timely down-regulation of IL-1 appears consistent with an inflammatory response that is sufficient to clear the bacterial infection with less corneal damage. PMID- 10843719 TI - Increased CD40 expression on muscle cells of polymyositis and dermatomyositis: role of CD40-CD40 ligand interaction in IL-6, IL-8, IL-15, and monocyte chemoattractant protein-1 production. AB - In polymyositis (PM)/dermatomyositis (DM), T cells infiltrate the muscle tissues and interact with muscle cells via cell surface molecules. Recently, myoblasts have been reported to express CD40, but little is known about the role of CD40 in myoblasts. In the present study we examined the expression and involvement of CD40 and CD40 ligand (CD40L) in the interaction between muscle cells and T cells in PM/DM. Immunohistochemical staining revealed that CD40 was expressed on muscle cells in five of five PM and four of five DM patients, and that infiltrating mononuclear cells (MNCs) expressed CD40L in all cases of PM/DM. These CD40L expressing MNCs were primarily CD4+ T cells. IFN-gamma, which is known to induce CD40 expression on various types of cells, was also expressed on the MNCs in four of the PM and four of the DM patients. Although cultured human myoblasts (SkMC 2859) did not express CD40 constitutively, IFN-gamma induced CD40 expression in a dose-dependent manner. To clarify the functional roles of CD40-mediated signals, the effects of a trimeric form of recombinant human CD40L on cytokine production were studied in SkMC 2859 that were prestimulated with IFN-gamma to express CD40. Recombinant human CD40L markedly increased the production of IL-6, IL-8, IL-15, and monocyte chemoattractant protein-1 of SkMC 2859. The expression of these humoral factors in muscle cells of PM and DM was demonstrated by immunohistochemistry. These results suggest that interaction between T cells and muscle cells via the CD40-CD40L system contributes to the immunopathogenesis of PM/DM by augmenting inflammation via cytokine production by the muscle cells. PMID- 10843720 TI - Human TNF can induce nonspecific inflammatory and human immune-mediated microvascular injury of pig skin xenografts in immunodeficient mouse hosts. AB - TNF activates endothelial cells to express cell surface molecules that are necessary to recruit a local infiltrate of leukocytes. Because the actions of this proinflammatory cytokine are not species restricted, we investigated whether human TNF can up-regulate porcine endothelial adhesion molecules to elicit human T cell infiltration and damage of pig skin xenografts in a chimeric immunodeficient mouse model. We have previously demonstrated the vigorous rejection of human skin allografts and the absence of injury to porcine skin xenografts in human PBMC-SCID/beige mice. Intradermal administration of human TNF at high doses (600 or 2000 ng) caused nonspecific inflammatory damage of pig skin grafts, whereas low concentrations of TNF (60 or 200 ng) resulted in human PBMC dependent injury of porcine endothelial cells. There was a strong correlation among pig skin xenograft damage, human T cell infiltration, and the TNF-induced up-regulation of swine MHC class I and class II molecules, VCAM-1, and, in particular, the de novo expression of porcine E-selectin. The microvascular damage and leukocytic infiltration elicited by TNF were enhanced by porcine IFN gamma, suggesting that xenografts may be less prone to cytokine-mediated injury due to the species-restricted effects of recipient IFN-gamma. Our results indicate that maintenance of a quiescent endothelium, which does not express E selectin or other activation-dependent adhesion molecules, is important in preventing human anti-porcine T cell xenoresponses in vivo and that TNF signaling molecules and TNF-responsive gene products are appropriate therapeutic targets to protect against human T cell-mediated rejection of pig xenografts. PMID- 10843721 TI - A self MHC class II beta-chain peptide prevents diabetes in nonobese diabetic mice. AB - We explored T cell responses to the self class II MHC (I-Ag7) beta-chain-derived peptides in diabetic and prediabetic nonobese diabetic (NOD) mice. We found that one of these immunodominant epitopes of the beta-chain of I-Ag7 molecule, peptide 54-76, could regulate autoimmunity leading to diabetes in NOD mice. T cells from prediabetic young NOD mice do not respond to the peptide 54-76, but T cells from diabetic NOD mice proliferated in response to this peptide. T cells from older nondiabetic mice or mice protected from diabetes do not respond to this peptide, suggesting a role for peptide 54-76-specific T cells in pathogenesis of diabetes. We show that this peptide is naturally processed and presented by the NOD APCs to self T cells. However, the peptide-specific T cells generated after immunization of young mice regulate autoimmunity in NOD mice by blocking the diabetogenic cells in adoptive transfer experiments. The NOD mice immunized with this peptide are protected from both spontaneous and cyclophosphamide-induced insulin dependent diabetes mellitus. Immunization of young NOD mice with this peptide elicited T cell proliferation and production of Th2-type cytokines. In addition, immunization with this peptide induced peptide-specific Abs of IgG1 isotype that recognized native I-Ag7 molecule on the cell surface and inhibited the T cell proliferative responses. These results suggest that I-Abetag7(54-76) peptide reactive T cells are involved in the pathogenesis of diabetes. However, immunization with this peptide at young age induces regulatory cells and the peptide-specific Abs that can modulate autoimmunity in NOD mice and prevent spontaneous and induced diabetes. PMID- 10843722 TI - Up-regulation of macrophage inflammatory protein-3 alpha/CCL20 and CC chemokine receptor 6 in psoriasis. AB - Autoimmunity plays a key role in the immunopathogenesis of psoriasis; however, little is known about the recruitment of pathogenic cells to skin lesions. We report here that the CC chemokine, macrophage inflammatory protein-3 alpha, recently renamed CCL20, and its receptor CCR6 are markedly up-regulated in psoriasis. CCL20-expressing keratinocytes colocalize with skin-infiltrating T cells in lesional psoriatic skin. PBMCs derived from psoriatic patients show significantly increased CCR6 mRNA levels. Moreover, skin-homing CLA+ memory T cells express high levels of surface CCR6. Furthermore, the expression of CCR6 mRNA is 100- to 1000-fold higher on sorted CLA+ memory T cells than other chemokine receptors, including CXCR1, CXCR2, CXCR3, CCR2, CCR3, and CCR5. In vitro, CCL20 attracted skin-homing CLA+ T cells of both normal and psoriatic donors; however, psoriatic lymphocytes responded to lower concentrations of chemokine and showed higher chemotactic responses. Using ELISA as well as real time quantitative PCR, we show that cultured primary keratinocytes, dermal fibroblasts, and dermal microvascular endothelial and dendritic cells are major sources of CCL20, and that the expression of this chemokine can be induced by proinflammatory mediators such as TNF-alpha/IL-1 beta, CD40 ligand, IFN-gamma, or IL-17. Taken together, these findings strongly suggest that CCL20/CCR6 may play a role in the recruitment of T cells to lesional psoriatic skin. PMID- 10843723 TI - CD40-ligated dendritic cells effectively expand melanoma-specific CD8+ CTLs and CD4+ IFN-gamma-producing T cells from tumor-infiltrating lymphocytes. AB - Professional APC, notably dendritic cells (DC), are necessary for stimulation and expansion of naive T cells. By means of murine models, the interaction between CD40 on DC and its ligand CD154 has been recognized as an important element for conditioning of DC to prime and expand CTL. We translated these findings into the human system, scrutinizing the ability of DC to initiate clonal expansion of single T cells. DC generated under completely autologous conditions from peripheral blood monocytes were cocultured at a rate of 0.3 cell/well with melanoma-infiltrating T cells; this procedure guaranteed that either a CD4+ or a CD8+ cell interacted with the DC, thus avoiding the contact of more than one T cell to the DC. In the absence of further stimulation, this cloning protocol yielded almost exclusively CD4+ T cell clones that predominantly exhibited a Th2 phenotype. However, cross-linking of CD40 on DC resulted in the induction of IFN gamma-producing Th1 CD4+ T cell clones. In addition, CD40-activated DC were capable of expanding CD8+ CTL clones. The ratio of CD4 to CD8 T cell clones corresponded to the ratio present in the initial tumor-infiltrating lymphocyte preparation. The CTL clones efficiently lysed autologous tumor cells whereas autologous fibroblasts or MHC-mismatched melanoma cells were not killed. Our findings support the critical role of CD40/CD154 interactions for the induction of cellular immune responses. PMID- 10843724 TI - Histamine is a potent inducer of IL-18 and IFN-gamma in human peripheral blood mononuclear cells. AB - Histamine (10-7 to 10-4 M) concentration-dependently stimulated the production of IL-18 and IFN-gamma and inhibited the production of IL-2 and IL-10 in human PBMCs. Histamine in the same concentration range did not induce the production of IL-12 at all. The stimulatory or inhibitory effects of histamine on cytokine production were all antagonized by H2 receptor antagonists ranitidine and famotidine in a concentration-dependent manner, but not by H1 and H3 receptor antagonists. Selective H2 receptor agonists, 4-methylhistamine and dimaprit, mimicked the effects of histamine on five kinds of cytokine production. The EC50 values of histamine, 4-methylhistamine, and dimaprit for the production of IL-18 were 1.5, 1.0, and 3.8 microM, respectively. These findings indicated that histamine caused cytokine responses through the stimulation of H2 receptors. All effects of histamine on cytokine responses were also abolished by the presence of either anti-IL-18 Ab or IL-1beta-converting enzyme/caspase-1 inhibitor, indicating that the histamine action is dependent on mature IL-18 secretion and that IL-18 production is located upstream of the cytokine cascade activated by histamine. The addition of recombinant human IL-18 to the culture concentration dependently stimulated IL-12 and IFN-gamma production and inhibited the IL-2 and IL-10 production. IFN-gamma production induced by IL-18 was inhibited by anti-IL 12 Ab, showing the marked contrast of the effect of histamine. Thus histamine is a very important modulator of Th1 cytokine production in PBMCs and is quite unique in triggering IL-18-initiating cytokine cascade without inducing IL-12 production. PMID- 10843725 TI - Recognition of sulfamethoxazole and its reactive metabolites by drug-specific CD4+ T cells from allergic individuals. AB - The recognition of the antibiotic sulfamethoxazole (SMX) by T cells is usually explained with the hapten-carrier model. However, recent investigations have revealed a MHC-restricted but processing- and metabolism-independent pathway of drug presentation. This suggested a labile, low-affinity binding of SMX to MHC peptide complexes on APC. To study the role of covalent vs noncovalent drug presentation in SMX allergy, we analyzed the proliferative response of PBMC and T cell clones from patients with SMX allergy to SMX and its reactive oxidative metabolites SMX-hydroxylamine and nitroso-SMX. Although the great majority of T cell clones were specific for noncovalently bound SMX, PBMC and a small fraction of clones responded to nitroso-SMX-modified cells or were cross-reactive. Rapid down-regulation of TCR expression in T cell clones upon stimulation indicated a processing-independent activation irrespective of specificity for covalently or noncovalently presented Ag. In conclusion, our data show that recognition of SMX presented in covalent and noncovalent bound form is possible by the same TCR but that the former is the exception rather than the rule. The scarcity of cross reactivity between covalently and noncovalently bound SMX suggests that the primary stimulation may be directed to the noncovalently bound SMX. PMID- 10843726 TI - Ro60 peptides induce antibodies to similar epitopes shared among lupus-related autoantigens. AB - The coexistence of autoantibodies to ribonucleoproteins (RNP) in sera of patients with systemic lupus erythematosus has been attributed to intermolecular determinant spreading among physically associated proteins. Recently, we showed that murine Ab responses to rRo60 or Ro60 peptides were diversified unexpectedly to small nuclear RNP. In this investigation, the mechanisms for this autoantibody diversification were examined. Intramolecular determinant spreading was demonstrated in mice immunized with human or mouse Ro60316-335. Immune sera depleted of anti-peptide Ab immunoprecipitated Ro60-associated mY1 and mY3 RNA and remained reactive to a determinant on Ro60128-285. Absorption with the immunogen depleted the immune sera completely of anti-Golgi complex Ab (inducible only with human Ro60316-335) and anti-La Ab, and reduced substantially Ab to SmD and 70-kDa U1RNP. Mouse rRo60 completely inhibited the immune sera reactivity to La, SmD, and 70-kDa U1RNP. However, La, SmD, and 70-kDa U1RNP preferentially inhibited the antiserum reactivities to these Ags, respectively. Affinity purified anti-La Ab were reactive with Ro60, La, SmD, and 70-kDa U1RNP. These results provide evidence that a population of the induced autoantibodies recognized determinants shared by these autoantigens. Lack of sequence homology between Ro60316-335 and La, SmD, or 70-kDa U1RNP suggests that these determinants are conformational. Interestingly, similar cross-reactive autoantibodies were found in NZB/NZW F1 sera. Thus, a single molecular mimic may generate Ab to multiple RNP Ags. Furthermore, cross-reactive determinants shared between antigenic systems that are not associated physically (Ro/La RNP and small nuclear RNP) may be important in the generation of autoantibody diversity in systemic lupus erythematosus. PMID- 10843727 TI - Alterations in the spinal cord T cell repertoire during relapsing experimental autoimmune encephalomyelitis. AB - The CNS T cell repertoire was analyzed by RT-PCR, spectratyping, and nucleotide sequencing of the amplified products at different times following adoptive transfer of a CD4+, Th1, VB2+ encephalitogenic SJL/J proteolipid protein peptide 139-151-specific T cell clone. The third complementarity-determining region of TCR B chains in the spinal cord was used as an indicator of T cell heterogeneity. Spectratypic analysis revealed that a single peak corresponding to the third complementarity-determining region of the initiating T cell clone predominated during the acute phase. During recovery and relapse the complexity of the spectratype increased. DNA sequence analysis revealed that the donor clone predominated at the acute phase. By the first relapse the donor clone, although represented most frequently, was a minority of the total. Spectratypic analysis of the same samples for several other VB families revealed their presence during acute disease or relapses but, with the exception of VB17, their absence during the recovery stage. PMID- 10843728 TI - Autocrine and paracrine regulation by cytokines and growth factors in melanoma. AB - Tumour development and progression involves the expression of oncogenes and inactivation of tumour suppressor genes, leading to the appearance of multiple malignant characteristics. Malignant melanoma cells express different growth factors and cytokines and their receptors in respective stages of tumour progression, which by autocrine and paracrine effects enable them to grow autonomously and confer competence to metastasis. Autocrine growth factors (bFGF, MGSA/GRO, IL-8 and sometimes IL-6, PDGF-A, IL-10) produced by melanoma cells stimulate proliferation of the producing cell itself, while paracrine growth factors (for example PDGF, EGF, TGF-beta, IL-1, GM-CSF, IGF-I, NGF, VEGF) modulate the microenvironment to the benefit of tumour growth and invasion. Paracrine effects include angiogenesis, stroma formation, modulation of host immune response, activation of proteolytic enzymes, adhesion or motility and metastasis formation. Some growth factors have inhibitory effects on melanocytes and early lesions (IL-1, IL-6, TGF-beta, OSM, TNF and IFN) but not on advanced stage melanomas, and in some cases they switch to autocrine stimulator (IL-6, TGF beta). Understanding the involvement of different growth factors and cytokines in the molecular mechanism of melanoma progression will help to provide an insight into new future therapeutic approaches for melanoma. PMID- 10843729 TI - The complete cDNA sequences of IL-2, IL-4, IL-6 AND IL-10 from the European rabbit (Oryctolagus cuniculus). AB - The cDNAs for four rabbit cytokine genes [interleukin 2 (IL-2), IL-4, IL-6 and IL 10] have been cloned from primary lymphocytes by polymerase chain reaction (PCR) methods. IL-2 and IL-10 are both highly conserved between rabbit and other species. IL-4 and IL-6 are less strongly conserved, at both nucleotide and amino acid levels, and exhibit structural differences. An extension of the coding region of rabbit IL-6 relative to all other reported IL-6 genes results from a mutation in the usual stop codon which allows translation to continue for a further 27 amino acids. Analysis of IL-6 from four other lagomorph species suggests that this mutation is specific to the European rabbit. Sequence and structural differences of IL-4 and IL-6, while presumably not altering function, may render them highly species-specific. Several alternatively spliced variants of IL-2 and IL-4 are also reported. PMID- 10843730 TI - Porcine interleukin 18: cloning, characterization of the cDNA and expression with the baculovirus system. AB - We have isolated and sequenced a cDNA that contains the coding sequence of porcine interleukin 18 (IL-18) and the recombinant protein of porcine IL-18 was expressed using the baculovirus system. The open reading frame (ORF) of the porcine IL-18 cDNA is 579 base pairs (bp) in length and encodes 192 amino acids. The predicted amino acid sequence is 76.7%, 64.7% and 61.6% homologous to the predicted human, murine and rat amino acid sequences, respectively. The porcine precursor and mature IL-18 protein were expressed respectively in Trichoplusia ni -derived (Tn5) cells using the baculovirus Autografha californica nuclear polyhedorosis virus (AcNPV) as a vector. Tn5 cells infected with recombinant virus containing a whole IL-18 protein coding region sequence secreted porcine precursor IL-18 into the culture medium. On the other hand, Tn5 cells infected with recombinant virus containing a mature IL-18 protein coding region sequence expressed several proteins in the cell lysates, but did not secrete mature protein into the culture medium efficiently. Immunoblotting analysis of recombinant protein showed cross-reactivity with anti-human IL-18 polyclonal antibody. The mature form of porcine IL-18 protein induced IFN-gamma production in suboptimal doses of anti-CD3 antibody and concanavalin A- (ConA) stimulated porcine peripheral blood mononuclear cells (PBMC), but the precursor form had little effect. PMID- 10843731 TI - Expression and purification of woodchuck tumour necrosis factor alpha. AB - The production of recombinant woodchuck cytokines is an essential prerequisite to study the immune response to hepadnavirus infection in the woodchuck model. Woodchuck tumour necrosis factor-alpha (TNF-alpha) was expressed in mammalian cells and in Escherichia coli. A test system for the biological activity of woodchuck TNF-alpha was established on basis of its cytotoxic effect to the murine fibrosarcoma cell line L929. Recombinant TNF-alpha was purified and used for the production of neutralizing antisera. PMID- 10843732 TI - IL-4-dependent induction of BCL-2 and BCL-X(L)IN activated T lymphocytes through a STAT6- and pi 3-kinase-independent pathway. AB - Both B and T lymphocytes require ongoing signals to maintain their viability. The pleiotropic cytokine interleukin (IL-) 4 plays an important role in the maintenance of activated T cells, perhaps reflecting induction of the anti apoptotic genes Bcl-2 and Bcl-X(L). However, it is not known which of the signalling pathways known to link the IL-4 receptor with transcription regulation are required, or if the levels of Bcl-2/X induction under such physiologic conditions are sufficient to account for the anti-apoptotic effects of IL-4. We report here that although blockade of pathways (PI 3-kinase and pp70 S6 kinase) recruited by the IRS-1/2 adaptor proteins inhibited the anti-apoptotic function of IL-4, Bcl-2/X induction were normal. These findings were recapitulated in primary and culture-adapted T cells whose Stat6 signalling pathway also was defective. These results demonstrate that both the Stat6 and PI 3-kinase pathways can be dispensable for Bcl-2/X induction by IL-4, thus suggesting the involvement of an additional signal transduction pathway. Moreover, the preservation of Bcl 2/X induction despite inhibition of the anti-apoptotic function of IL-4 indicates that this cytokine activates additional protective mechanisms. PMID- 10843733 TI - Transcriptional activation of indoleamine dioxygenase by interleukin 1 and tumor necrosis factor alpha in interferon-treated epithelial cells. AB - Interferon (IFN)-gamma-induced indoleamine 2,3-dioxygenase (IDO) activity is enhanced synergistically by interleukin (IL-)1, tumor necrosis factor-alpha (TNF alpha) and LPS in IFN-treated macrophages by increasing IDO mRNA concentration. These studies demonstrate that IFN-treated HeLa cells also exhibit dose-dependent enhancement of IDO induction by TNF-alpha and IL-1, with maximal effects at concentrations of 5 ng/ml and 3 ng/ml, respectively. Furthermore, with sub optimal IFN concentrations, cells treated with maximally effective concentrations of TNF-alpha or IL-1alpha required 3-5 times less IFN to induce the same level of IDO activity as that observed with IFN alone. To detect changes in transcriptional activation of the IDO gene, HeLa cells were transfected with a plasmid containing the IDO 5' regulatory region upstream of a green fluorescent protein (GFP) reporter gene. In transfected cells, IFN induced both IDO activity and GFP that was detected by flow cytometry. When cell-sorted, transfected cells were stimulated with IFN in combination with TNF-alpha or IL-1 but not LPS, increased GFP was detected in comparison to transfected cells treated with IFN alone. Furthermore, increases in GFP expression correlated with IDO enzymatic activity, indicating that combinations of IFN with IL-1 or TNF-alpha increase the transcriptional activity of the IDO promoter region. PMID- 10843734 TI - Divergent cell-specific effects of activin-A on thymocyte proliferation stimulated by phytohemagglutinin, and interleukin 1beta or interleukin 6 in vitro. AB - Activin-A is a member of the transforming growth factor-beta (TGF-beta) cytokine family. Based on studies in several cell systems, activin-A has been postulated to be a specific inhibitor of the actions of the inflammatory cytokine, interleukin 6. In cultures of adult rat thymocytes, activin-A inhibited sub optimal phytohemagglutinin-induced and interleukin 1beta-stimulated proliferation, as measured by [(3)H]-thymidine incorporation in vitro. In contrast with TGF-beta1, which exerted similar inhibitory effects on thymocyte proliferation, activin-A activity was reduced by increasing the concentration of phytohemagglutinin or addition of the reducing agent, beta-mercaptoethanol. Both activin-A and TGF-beta1 inhibited the in vitro production of interleukin 6 by thymocytes in the presence of phytohemagglutinin and interleukin 1beta. In the presence of exogenous interleukin 6, however, both activin-A and TGF-beta1 stimulated thymocyte proliferation. These data suggest that activin-A inhibits thymocyte growth and differentiation, at least in part, by inhibiting endogenous production of interleukin 6, but stimulates thymocyte growth when exogenous interleukin 6 is present in vitro. These data indicate that activin interacts with other cytokines to exert complex regulation of T cell development, and is not an inhibitor of interleukin 6 action in all cell systems. PMID- 10843735 TI - Interferon-gamma and interleukin 4 inhibit interleukin 1beta-induced delayed prostaglandin E(2)generation through suppression of cyclooxygenase-2 expression in human fibroblasts. AB - Interleukin (IL-)1 stimulates prostaglandin E(2)(PGE(2)) generation in fibroblasts, and preferential couplings between particular phospholipase A(2)(PLA(2)) and cyclooxygenase (COX) isozymes are implicated with IL-1-induced delayed PGE(2)generation. The regulatory effects of interferon (IFN)-gamma and IL 4 on IL-1beta-induced COX, PLA(2)isoforms expression and terminal delayed PGE(2)generation were examined in three types of human fibroblasts. These human fibroblasts constitutively expressed cytosolic PLA(2)(cPLA(2)) and COX-1 enzymes, and exhibited delayed PGE(2)generation in response to IL-1beta. IL-1beta also stimulated expression of cPLA(2)and COX-2 only, while constitutive and IL-1beta induced type IIA and type V secretory PLA(2)s (sPLA(2)s) expression could not be detected. A COX-2 inhibitor and cPLA(2)inhibitor markedly suppressed the IL-1beta induced delayed PGE(2)generation, while a type IIA sPLA(2)inhibitor failed to affect it. IFN-gamma and IL-4 dramatically inhibited the IL-1beta-induced delayed PGE(2)generation; these cytokines apparently suppressed IL-1beta-stimulated COX-2 expression and only weakly suppressed cPLA(2)expression in response to IL-1beta. These results indicate that IL-1beta-induced delayed PGE(2)generation in these human fibroblasts mainly depends on de novo induction of COX-2 and cPLA(2), irrespective of the constitutive presence of COX-1, and that IFN-gamma and IL-4 inhibit IL-1beta-induced delayed PGE(2)generation by suppressing, predominantly, COX-2 expression. PMID- 10843736 TI - Stimulation of osteoclast differentiation in vitro by mouse oncostatin M, leukaemia inhibitory factor, cardiotrophin-1 and interleukin 6: synergy with dexamethasone. AB - The role of oncostatin M in bone metabolism is not clearly defined, and the actions of mouse oncostatin M (mOSM) on osteoclast development has not been previously determined. We therefore examined the ability of recombinant mOSM to stimulate osteoclast formation and activity using cocultures of murine calvaria and bone marrow cells, and compared the responses to other members of the interleukin 6 family of cytokines including mouse leukaemia inhibitory factor (LIF), cardiotrophin-1 (CT-1) and IL-6. Mouse OSM, LIF and CT-1 strongly induced the formation of tartrate resistant acid phosphatase positive (TRAP(+)) multinucleated cells (MNC) in a dose-dependent fashion. OSM, LIF or CT-1 also elevated the number and size of resorptive pits when cocultures were added to smooth cortical bone slices, indicating enhancement of osteoclast activity. The activity of OSM was reduced by indomethacin (10(-8)-10(-6) M), whereas addition of dexamethasone (DEX) at 10(-7)-10(-5) M synergistically enhanced OSM-induced numbers of TRAP(+)MNC. DEX (10(-7) M) costimulation also synergistically enhanced TRAP(+)cell numbers of LIF, and CT-1 treated cocultures. IL-6 had no activity alone, but further enhanced TRAP(+)cell formation in mOSM or DEX (10(-7) M) treated cocultures. When added to mouse calvarial osteoblast cultures, mOSM induced secretion of IL-6 protein and elevation of mRNA whereas LIF or CT-1 did not. IL-6 mRNA levels and protein secretion were reduced in osteoblasts by costimulation with DEX. These results show that mouse OSM, LIF and CT-1 induce osteoclast differentiation and activation, that DEX synergizes with each in this activity, and that mouse OSM induces responses in osteoblasts that are not shown by LIF or CT-1. Collectively these data suggest an important role of these cytokines in osteoporosis caused by high levels of corticosteroid. PMID- 10843737 TI - Expression and regulation of histidine decarboxylase mRNA expression in the uterus during pregnancy in the mouse. AB - It has been hypothesized that hormonally regulated histamine production plays a role in preparation of the uterus for implantation. Histidine decarboxylase (HDC) is the rate-limiting enzyme for histamine production. The current study was designed to determine intrauterine expression of HDC mRNA expression during pregnancy in the mouse. High levels of HDC mRNA expression were observed in the preimplantation mouse uterus with peak expression occurring on day 4. High levels of HDC mRNA expression were also detected in the post-implantation uterus. In an effort to determine whether HDC mRNA is regulated by pro-inflammatory cytokines, the HDC mRNA pattern was compared to intrauterine expression of mRNA's for interleukin-1alpha (IL-1alpha), IL-1beta, macrophage chemotactic protein-1 (MCP 1) and RANTES (regulated on activation, normal T expressed and secreted) during the peri-implantation period. IL-1beta, MCP-1 and RANTES mRNA levels were increased in the uterus on days 1-2 and on days 4-5. Increased expression of IL 1alpha mRNA was observed on days 1-2 and days 5-7. There was no clear relationship between HDC mRNA expression and cytokine/chemokine mRNA expression. Progesterone-stimulated intrauterine expression of HDC mRNA. Intrauterine cytokine/chemokine mRNA was also hormonally regulated. This data allowed the possibility that one or more of these pro-inflammatory cytokines could be involved in regulating intrauterine HDC mRNA production. Recombinant IL-1alpha, IL-1beta, MCP-1 and RANTES all failed to induce HDC mRNA expression in the preimplantation uterus in a mouse pseudopregnancy model. At the same time, IL 1beta induced the expression of mRNA for each of the four cytokines/chemokines. Despite the fact that these were also produced in the uterus during pregnancy and were hormonally regulated, none of these cytokines induced intrauterine HDC mRNA expression. The data suggest that progesterone is involved in the regulation of HDC mRNA expression in the preimplantation uterus, but IL-1alpha/beta, MCP-1 and RANTES, which have been reported to regulate histamine synthesis during inflammatory processes, do not appear to play a role. PMID- 10843738 TI - Long-term overexpression of human granulocyte colony-stimulating factor in transgenic mice: persistent neutrophilia with no increased mortality for more than one year. AB - To investigate possible adverse consequences of persistent neutrophil overproduction, mice transgenic for human granulocyte colony-stimulating factor (hG-CSF) were studied for more than 1 year. They showed marked granulocytopoiesis and neutrophilia. Continuous medullary and extramedullary granulocytopoiesis resulted in marked changes in bone and liver. In the liver, haemorrhage and focal necrosis and a few haemangiosarcomas were present, presumably caused by the destructive granulocytopoiesis. Despite the high incidence of lung infiltration by mature neutrophils, lung lesions rarely appeared. Although there was a persistent increase in neutrophils, mortality of the mice did not differ from that of non-transgenic littermates at least within 1 year after birth. Factors other than overproduction of G-CSF and extensive neutrophilia could be required for the development of neutrophil-mediated acute and chronic tissue damage. PMID- 10843739 TI - Expression of IL-8 gene in human monocytes and lymphocytes: differential regulation by TNF and IL-1. AB - TNF-alpha and IL-1 were reported to be the most powerful inducers of IL-8 in a multitude of cells, including leukocytes. In this study, we investigated TNF alpha- and IL-1-mediated regulation of IL-8 gene expression in non-fractionated PBMC, and purified monocyte (MO) and lymphocyte (LY) fractions. Our analysis revealed that purified human MO did not respond to exogenous TNF-alpha with the induction of IL-8 mRNA or protein, nor require endogenous TNF-alpha for IL-8 expression. In contrast, in the presence of exogenous IL-1alpha and IL-1beta a substantial enhancement of IL-8 mRNA and protein expression in MO was observed. Nevertheless, antibodies to IL-1alpha and IL-1beta were unable to downregulate the expression of IL-8 in resting adherent or Staphylococcus aureus Cowan 1 (SAC) stimulated MO. In contrast with MO, purified LY and non-fractionated PBMC expressed IL-8 in response to exogenous TNF-alpha, similar to exogenous IL-1alpha and IL-1beta. As was seen with MO, antibodies to TNF-alpha, IL-1alpha and IL 1beta did not inhibit the expression of IL-8 in purified LY and non-fractionated PBMC stimulated with SAC and LPS. Taken together, our data demonstrate major differences in responsiveness of MO and LY to exogenous TNF-alpha and IL-1, and suggest relative autonomy of IL-8 gene expression in these cells that does not require accessory cytokines but can be induced directly by exogenous stimuli. PMID- 10843740 TI - Production of tumour necrosis factor alpha by primary cultured rat alveolar epithelial cells. AB - Tumour necrosis factor alpha(TNF-alpha) is one of the most important pro inflammatory cytokines, which plays an important role in host defense and acute inflammation related to tissue injury. The major source of TNF-alpha has been shown to be immune cells such as macrophages and neutrophils. In the present study, we demonstrated that LPS-treatment on alveolar epithelial cells isolated from adult rat lungs also induced a dose- and time-dependent release of TNF alpha. The purity and identity of these cells were examined by immunofluorescent staining and confocal microscopy with antibodies for cytokeratin and pro surfactant protein C, markers for epithelial cells and type II pneumocytes respectively. Positive staining of TNF-alpha was observed throughout the cell layer and localized intracellularly. LPS-induced TNF-alpha production from alveolar epithelial cells was blocked not only by cycloheximide, an inhibitor of protein translation, but also by actinomycin D, an inhibitor of gene transcription. The mRNA of TNF-alpha rapidly increased within 1 h of LPS stimulation. These data suggest that LPS-induced TNF-alpha production from alveolar epithelial cells is primarily regulated at the transcriptional level, which is different from that of macrophages and neutrophils. TNF-alpha produced by alveolar epithelial cells may function as an alert signal in host defense to induce production of other inflammatory mediators. PMID- 10843741 TI - Interleukin 6 promotes vasculogenesis of murine brain microvessel endothelial cells. AB - Interleukin 6 (IL-6) is a cytokine that acts on a wide range of tissues influencing cell growth and differentiation. Here we show that IL-6 plays a role in the early vascular development (vasculogenesis) in the central nervous system (CNS). We report that IL-6 induces the proliferation of brain microvascular endothelial cells in vitro. Furthermore, IL-6 significantly accelerates the formation of tube-like structures by these cells in Matrigel basement matrix. Moreover, IL-6 mRNA is expressed in vivo in two physiological conditions in which vascularization in the CNS is important: (1) during normal brain development, (2) during the healing process of a traumatic brain injury. Expression of IL-6 mRNA coincides with the expression of vascular endothelial growth factor (VEGF) mRNA in the developing brain with decreasing expression following birth. However, IL-6 mRNA can be detected in the healing adult murine brain tissue by in situ hybridization coinciding with the period of intense tissue reorganization. The transient upregulation of IL-6 mRNA during normal brain development and at brain injury site and the effect of IL-6 on in vitro vasculogenesis suggest that IL-6 may play a role in normal physiology of vascularization in the CNS. PMID- 10843742 TI - A single injection of polyethylene-glycol granulocyte colony-stimulating factor strongly prolongs survival of mice with systemic candidiasis. AB - Systemic candidiasis is a life-threatening disease occurring in immunocompromized patients. Granulocyte colony-stimulating factor (G-CSF) reduces mortality in experimental invasive candidiasis. Covalent conjugation of polyethylene-glycol (peg) to proteins increases their stability and in vivo bioactivity. In this study, the effect of a single subcutaneous injection of peg-G-CSF on lethal candidiasis was assessed. This was performed in acute and chronic candidiasis models in non-neutropenic FVB/N mice. Peg-G-CSF rapidly increased circulating polymorphonuclear leukocyte (PMNL) numbers in mice, sustaining high for >4 days. Candida albicans outgrowth from kidneys of infected mice was strongly reduced after peg-G-CSF treatment (5.76 log cfu/g kidney vs 7.66 control), with absence of hyphal outgrowth and enhanced PMNL influx. Moreover, peg-G-CSF increased survival of C. albicans -infected mice, whereas efficacy of uncoupled G-CSF was obtained only after repeated treatment. These data document a potent in vivo biological effect of peg-G-CSF, resulting in strongly enhanced resistance against systemic candidiasis. PMID- 10843743 TI - Diminished interleukin 6 (IL-6) production during scarless human fetal wound repair. AB - Fetal wound healing is characterized by minimal inflammation and scarless repair. IL-6 stimulates inflammation in postnatal wound healing. We hypothesized that fetal skin has a diminished IL-6 response and that exogenous IL-6 will result in scar formation. Human adult or fetal skin was placed subcutaneously in SCID mice and incisionally wounded. Wounds were excised after 4, 12, 24 or 72 h for IL-6 mRNA quantification by RT-PCR. In other grafts, 5 microgram of IL-6 was injected at wounding and then harvested at 7 days for analysis of scar formation. IL-6 production was examined in primary cultures of human fetal or adult dermal fibroblasts incubated for 8 h with 0, 0.1, 1 or 10 ng/ml of PDGF-BB. IL-6 mRNA was detected 4 h after wounding in fetal and adult wounds, but by 12 h there was no IL-6 mRNA in the fetal wounds. Adult wounds had IL-6 mRNA persisting to 72 h. IL-6 administration to fetal wounds resulted in scar formation. Fetal fibroblasts produced less IL-6 protein and mRNA at all points examined (P<0.01 vs adult). Diminished production of inflammatory cytokines such as IL-6 may be responsible for the lack of inflammation seen during fetal wound healing. Diminished inflammation may provide a permissive environment for scarless wound healing. PMID- 10843744 TI - Circulating transforming growth factor beta(1) as an indicator of hepatic function impairment in liver cirrhosis. AB - In the liver, transforming growth factor (TGF) -beta(1)is primarily responsible for activation of fat-storing cells, which are the main source of extracellular matrix proteins. Their deposition play a key role in the development of liver cirrhosis. The aim of this study was to evaluate plasma TGF-beta(1)in patients with different stages of liver cirrhosis and its possible use as an indicator of liver function impairment. TGF-beta(1)was measured in the plasma of 40 patients with liver cirrhosis. To estimate possible effect of liver insufficiency on plasma TGF-beta(1), patients were divided into three groups: A, B and C, univocal with Child-Pugh classes. Normal values were collected from 13 healthy volunteers. Liver cirrhosis resulted in a significant increase of plasma concentration of TGF beta(1)(39.3+/-3.8 ng/ml), which doubled normal values (18.3+/-1.6 ng/ml). The highest concentrations were observed in alcoholic patients (44.4+/-4.7 ng/ml). TGF-beta(1)level increased depending on the degree of liver insufficiency, demonstrated by a significant positive correlation with Child-Pugh score (r=0.591). Values in group A were similar to normal, but were significantly elevated in groups B and C. These findings suggest possible use of plasma TGF beta(1)measurement as an indicator of liver function impairment and possible marker of hepatic fibrosis progression in cirrhotic patients. PMID- 10843745 TI - Short-term treatment of relapsing remitting multiple sclerosis patients with interferon (IFN)-beta1B transiently increases the blood levels of interleukin (IL)-6, IL-10 and IFN-gamma without significantly modifying those of IL-1beta, IL 2, IL-4 and tumour necrosis factor-alpha. AB - We have studied the impact of short-term treatment with interferon (IFN)-beta1b of relapsing remitting (RR) multiple sclerosis (MS) patients' blood levels of type 1 and type 2 cytokines such as IFN-gamma, interleukin (IL)-1beta, IL-2, IL 4, IL-6, IL-10 and tumour necrosis factor (TNF)-alpha. These cytokines were measured by solid-phase ELISA. Serum samples were obtained prior to, and 2 and 12 hours after beginning of the treatment and 48 h after the last of 5 s.c. injections with 8 million IU IFN-beta1b given on alternate days for 10 days. The treatment was found to increase the circulating levels of IL-2, IL-6, IL-10 and IFN-gamma at some of the time points considered, with the effect acquiring statistical significance for IL-6, IL-10 and IFN-gamma. The blood levels of IL 1beta, IL-4 and TNF-alpha remained below the limit of sensitivity of the assays at any of the time points considered. If this in vivo study mirrors the impact of IFN-beta1b on MS patients' immune cells, these data demonstrate an activation of the immune system upon early treatment with the drug that does not lead to either type 1 or type 2 cytokine prevalence. PMID- 10843746 TI - Production of IL-10 and IL-12 in CD40 and interleukin 4-activated mononuclear cells from patients with Graves' disease. AB - We investigated the effect of T cell-dependent B cell activation on the production of IL-10 and IL-12 by peripheral blood mononuclear cells (PBMCs) obtained from patients with Graves' disease vs Hashimoto's thyroiditis, type 1 diabetes or normal controls. Incubation of PBMCs, from each of the subject groups, with a combination of anti-CD40 monoclonal antibodies and interleukin 4 (IL-4)-activated B cells, as shown by an increased level of soluble CD23. There was also a notable increase in the number of CD23(+)cells in PBMCs from patients with Graves' disease as compared to the other subject groups. This combination of B cell stimulants increased production of IL-10 in PBMCs obtained from patients with Graves' disease relative to those patients with Hashimoto's thyroiditis, type 1 diabetes, or the control subjects. The production of IL-12 showed wide variation that depended on the basal IL-12 level. In subjects with a low basal IL 12 level there was a positive correlation between the production of IL-12 and that of IL-10 from PBMCs stimulated with anti-CD40 antibodies plus IL-4. On the contrary, in the patients with a high basal IL-12 level, no change or a decrease of IL-12 production was observed after the stimulation. Thus, T cell-dependent B cell activation via a CD40 pathway triggers the overproduction of IL-10 and overcome the effect of IL-12 to shift the Th(1)/Th(2)balance to Th(2)dominance in patients with Graves' disease but not in Hashimoto's thyroiditis or type 1 diabetes. PMID- 10843747 TI - Effects of an intravenous catheter on the local production of cytokines and soluble cytokine receptors in healthy men. AB - Like many aspects of physiology, functions of the immune system show considerable diurnal variation. Studies investigating diurnal variations in the circulating amounts of cytokines, in general, used blood samples obtained from an intravenous catheter. The results of such studies may be confounded by an effect of the catheter on local cytokine production. We measured the levels of IL-6, tumour necrosis factor alpha (TNF-alpha) and soluble TNF receptors (sTNF-R) p55 and p75 in 20 healthy men between 09:00 and 19:00 h in plasma samples obtained from an intravenous catheter and in one additional sample obtained by a simple needle stick in the contralateral arm 10 h after baseline. In plasma from the catheter the levels of IL-6 increased significantly over time, TNF-alpha levels slightly decreased and the time courses of TNF receptor levels showed significant trends of a higher order. Control levels of IL-6, TNF-alpha and sTNF-R p75 measured in plasma obtained by needle stick after 10 h did not differ from baseline, and those of sTNF-R p55 were even higher. We conclude that local alterations in the production of cytokines and soluble cytokine receptors induced by an intravenous catheter represent an important confounding factor for studies investigating diurnal variations in immune functions. PMID- 10843749 TI - Engineered acid-stabile human interferon gamma. AB - Loss of anti-viral potency upon pH2-treatment is an inherent feature of interferon (IFN)-gamma. The phenomenon seems to be caused by dissociation of IFN gamma homodimer into subunits upon acidification and subsequent self-association of monomers into aggregates with reduced activity after neutralization. We demonstrated that acid-stability could be engineered into human IFN-gamma without affecting its specific activity. An artificial intra-monomer disulphide bond E7C/S69C stabilizes the dimeric form of the cytokine, which retained its full bioactivity after exposure to pH2. Acidification did not modify the antigenic structure of IFN-gamma as proved by a panel of mouse anti-human IFNgamma antibodies. PMID- 10843748 TI - SCF, IL-1beta, IL-1ra and GM-CSF in the bone marrow and serum of normal individuals and of AML and CML patients. AB - This study compared cytokine transcript and protein levels in BM cells of normal individuals and leukemic patients. AML differed from normal in that: (1) AML marrow cells contain more IL-1beta protein than normal cells, (2) IL-1ra transcripts are absent from AML marrow cells, (3) AML marrow serum contains less IL-1ra protein than normal, (4) peripheral blood and marrow serum of AML patireents contains more SCF protein than normal serum, and (5) SCF transcripts have been detected in AML marrow biopsies and not in aspirate cells. These data suggest that unbalanced cytokine production may make a significant contribution to the abnormal behaviour of AML cells. PMID- 10843750 TI - Internalization and nuclear translocation of IFN-gamma and IFN-gammaR: an ultrastructural approach. AB - IFN-gamma signalling involves the Jak-STAT pathway. However, several hypothesis have been proposed where the receptor and its ligand itself took an active role within the cell. Using a quantitative immunogold approach, we found that both IFN gamma and its receptor are rapidly internalized and translocated in the nucleus. We found that cell surface heparan sulfate, which binds IFN-gamma, delayed the nuclear accumulation of IFN-gamma suggesting that these molecules serve as storage depot around the cell for local delivery of the cytokine. PMID- 10843751 TI - In vivo subcellular target compartments of interferon-gamma and interferon-gamma receptor (alpha- and beta-chains) in mouse liver. AB - IFN-gamma displays several effects on different tissues via its specific cell surface receptor (IFN-gammaR). In order to identify the target compartments of IFN-gamma and IFN-gammaR (alpha and beta-chains), we used a quantitative immunogold approach. In physiological conditions, IFN-gamma and IFN-gammaR immunoreactivities were detected in the plasma membrane, in the endoplasmic reticulum area, in the mitochondria and in the nucleus. After a single IFN-gamma injection, we observed, in a quantitative manner, an increase of signal density without modification of the subcellular distribution of IFN-gamma and IFN-gammaR subunits. PMID- 10843752 TI - Cytokine-mediated differential regulation of macrophage activator protein-1 genes. AB - The regulation of macrophage activator protein-1 (AP-1) gene expression by LPS and cytokines is of potentially crucial importance in the pathogenesis of several diseases. The action of LPS and four cytokines on AP-1 gene expression in the murine macrophage J774.2 cell line was, therefore, studied. Exposure of the cells to IL-6 produced no changes in the mRNA levels of all AP-1 members studied. In contrast, the expression of JunB, c-jun and c-fos, but not JunD, was increased by LPS, TNF-alpha, IFN-gamma and IL-1, albeit with different kinetics and magnitude of induction. Electrophoretic mobility shift assays showed a close correlation between the expression of the AP-1 genes and the functional AP-1 DNA binding activity and, additionally, demonstrated the participation of heterodimeric interactions between the different members. These studies provide insights into the potential mechanisms that may be involved in the mediator-specific modulation of AP-1 regulated macrophage gene expression. PMID- 10843753 TI - The effect of interleukin-6 (IL-6)/gp130 signalling on biliary epithelial cell growth, in vitro. AB - The effect of IL-6 on the growth of mouse biliary epithelial cells (BEC), in vitro, was tested by comparing BEC obtained IL-6-deficient mice (IL-6(-/-)) to wild-type littermate controls (IL-6(+/+)), in two different media: simple serum free media (S-SFM), and complete serum-free media (C-SFM) containing forskolin, which stimulates BEC IL-6 production. In S-SFM, neither IL-6(+/+)nor IL-6(-/-)BEC constitutively produced IL-6 mRNA or protein, and there was no difference between IL-6(+/+)and IL-6(-/-)BEC growth. In contrast, when the BEC were maintained in C SFM, over 48 h, the growth of IL-6(+/+)BEC was 40% greater than IL-6(-/-)BEC (P<0.006). Enhanced IL-6(+/+)BEC growth in C-SFM was associated with induced expression of IL-6 mRNA and IL-6 protein secretion into the medium, upregulation of the IL-6Ralpha (gp80) and phosphorylation of the signal transducing molecule gp130. In C-SFM, anti-IL-6 neutralizing antibodies blocked enhanced IL-6(+/+)BEC growth, whereas exogenous rhIL-6 stimulated retarded growth of IL-6(-/-)BEC. Thus, under conditions that mimic an inflammatory or stressful microenvironment in vivo, BEC produce, secrete and respond to IL-6, via upregulation and activation of the IL-6Ralpha (gp80)/gp130 signaling system in an autocrine/paracrine manner. PMID- 10843754 TI - A functional role for interleukin (IL)-4-driven cyclic amp accumulation in human b lymphocytes. AB - Interleukin-4 (IL-4) regulates the expression of the 55-kDa alpha-subunit (CD25) of the IL-2 receptor complex in human B lymphocytes. This report suggests that the cAMP/protein kinase A (PKA) component of the IL-4 receptor signalling programme in human tonsillar B cells has a functionally important role in regulating expression of the CD25 gene by attenuating activity of a protein binding to a potent negative regulatory element (NRE) in the CD25 promoter; this effect can be mimicked by agents that elevate cAMP and blocked by inhibitors of PKA but not protein kinase C (PKC). In a B-cell line that fails to elevate cAMP, attenuate NRE-binding protein (NRE-BP) activity or express CD25 following IL-4 treatment, stimulation of cAMP accumulation by forskolin facilitates IL-4 mediated induction of both the endogenous gene and an exogenous reporter gene under the control of a minimal promoter/enhancer fragment of the CD25 gene. PMID- 10843755 TI - Expression and activity of 2-5A synthetase in the course of differentiation and apoptosis of PC12 cells. AB - The role of IFN-induced 2-5A system in cell differentiation has not been elucidated. While studying differentiation of PC12 cells we found that the simultaneous treatment of cells with NGF and IFN-gamma in serum-containing medium resulted first in the extension of neurites and then apoptosis. On the contrary, in serum-free medium the cells underwent a more rapid neuronal differentiation. Only the doses of NGF which induced the outgrowth of neurites from the cells were able to induce rapid cell death in combined treatment. When the cells were treated subsequently with NGF and IFN-gamma, the induction of death was observed with NGF post-treatment, but not with NGF pretreatment. Relying on these alternative biological responses, we studied the changes in 2-5A synthetase activity and its 43 kDa isoform expression in the course of differentiation and death of PC12 cells. The results of the present work showed that NGF-induced differentiation of the cells did not evoke any increase in 2-5A synthetase activity or any increase in the expression of its 43 kDa isoform. Moreover, the obtained results demonstrated that NGF could not significantly affect the IFN induced signalling pathway leading to the activation of 2-5A synthetase gene, at least regarding the studied enzyme activity. PMID- 10843756 TI - Expression of soluble CD137 correlates with activation-induced cell death of lymphocytes. AB - CD137 is a member of the tumour necrosis factor receptor family which delivers a potent co-stimulatory signal to T lymphocytes. Soluble forms of CD137 (sCD137) can be found at enhanced levels in sera of patients with rheumatoid arthritis. Here we show that expression of sCD137 lags behind that of membrane-bound CD137 (mCD137) by about 24 h. In fully activated lymphocytes, time of maximum increase and level of expression of sCD137 were at day 2 and 3, respectively. Expression of sCD137 in lymphocytes requires strong activation, and levels of sCD137 correlate negatively with lymphocyte proliferation and positively with the degree of activation-induced cell death caused by mitogen overstimulation. Since activation of lymphocytes through membrane-bound CD137 delivers a potent stimulatory signal, sCD137 may provide a negative control mechanism for immune responses. PMID- 10843757 TI - A role for potassium in TNF-induced apoptosis and gene-induction in human and rodent tumour cell lines. AB - Rat/mouse T cell hybridoma-derived PC60 R55/R75 cells were used as a model to study the role of intracellular potassium in TNF-induced apoptosis and gene induction. A reduction of intracellular potassium with nigericin or valinomycin (ionophores), or ouabain (Na(+)/K(+)-ATPase inhibitor) sensitized PC60 R55/R75 cells to TNF-induced apoptosis. TNF-induced GM-CSF release in PC60 R55/R75 cells was enhanced by nigericin or ouabain. Similar results were obtained with human cervix carcinoma cells HeLaH21 exposed to TNF. These results suggest a role for intracellular potassium in TNF-induced apoptosis and gene induction. PMID- 10843758 TI - Interleukin 15 is a growth factor for human thymocytes with preferential effect on CD8(+)cells. AB - The effects of IL-15, as compared to IL-2, on growth of human thymocytes has been evaluated. Expression of comparable amounts of receptor chains was found on IL-2 and IL-15 cultured thymocytes, as well as comparable receptor signalling. However, IL-15 was superior to IL-2 in promoting CD8(+)thymocyte growth, whereas CD4(+)cells proliferated to a higher extent in IL-2 cultures. CD4(+)8(+)and CD4( )8(-)thymocytes expanded equally well in both culture types. PMID- 10843759 TI - Combinations of the cytokines IL-12, IL-2 and IFN-alpha significantly augment whereas the cytokine IL-4 suppresses the cytokine-induced antibody-dependent cellular cytotoxicity of monoclonal antibodies 17-1A and BR55-2. AB - Since some cytokines effectively enhance the cytotoxicity of monoclonal antibodies, we investigated whether a combination of cytokines can augment the antibody-dependent cellular cytotoxicity (ADCC) of monoclonal antibodies 17-1A and BR55-2 against the colorectal carcinoma cell line HT29. Since monocytes/macrophages are important effector cells for ADCC, we used a new flow cytometric cytotoxicity assay, which allows the analysis of long-term-ADCC exerted by these cells. In our previous studies with peripheral blood mononuclear cells from normal donors, we found that IL-2, IL-12 and IFN-alpha increase ADCC. Therefore, we examined whether combination of these three cytokines with IL-2, IL 4, IL-6, IL-10, IL-12, IFN-alpha, IFN-gamma, GM-CSF, M-CSF and TNF-alpha may yield higher ADCC than obtained by the application of single cytokines. Indeed, we found that the combinations IL-2/IFN-alpha, IL-2/IL-12 and IL-12/IFN-alpha potentiated ADCC. Interestingly, the ineffective single cytokines TNF-alpha and GM-CSF in the combinations IL-2/TNF-alpha, IFN-alpha/TNF-alpha and IFN-alpha/GM CSF also proved to enhance ADCC. In contrast, IL-4 significantly suppressed the IL-2, IL-12 and IFN-alpha-induced ADCC. In addition, the immunosuppressive cytokine IL-10 in higher concentrations significantly suppressed the IL-12 induced-ADCC. Our results may be useful to find combinations of cytokines and mAb for the treatment of cancer. PMID- 10843761 TI - Transforming growth factor beta1 blocks the release of collagen fragments from boving nasal cartilage stimulated by oncostatin M in combination with IL-1alpha. AB - Oncostatin M in combination with interleukin-1 (IL-1) induced a rapid and reproducible release of collagen from bovine nasal cartilage in culture. This release was accompanied by a high collagenolytic activity and low or absent tissue inhibitor of metalloproteinase-1 activity in the culture medium. Transforming growth factor-beta1 was able to block this release of collagen from the tissue, and reduce the expression and secretion of collagenases whilst maintaining TIMP-1 levels from bovine nasal chondrocytes. This study shows for the first time that TGF-beta1 can protect cartilage collagen from destruction. PMID- 10843760 TI - Procalcitonin and calcitonin gene-related peptide decrease LPS-induced tnf production by human circulating blood cells. AB - The pathogenesis of septic shock is mainly due to unregulated tumour necrosis factor-alpha (TNF-alpha) production. Procalcitonin (PCT) and calcitonin gene related peptide (CGRP) are alternative transcription products of the calcitonin gene. Since high PCT levels have been described in human sepsis, and since CGRP inhibits TNF synthesis in rats, we examined the role of these peptides in the regulation of the inflammatory response during septic shock. LPS-induced TNF production was assessed using a human whole blood model. In this model, PCT (10( 7) M) and CGRP (10(-6) M) significantly inhibit TNF production by 27 and 24 % respectively. The effect of CGRP was reversed by CGRP 8-37 (10 microM), an antagonist of CGRP receptor. No effect on interleukin (IL)-1, IL-6 and IL-8 was found. This is the first description of an anti-inflammatory role for PCT and CGRP in humans. PMID- 10843762 TI - The immunosuppressant steroid cholesterylphosphoserine inhibits tumour necrosis factor-alpha secretion in vitro and in vivo. AB - The synthetic steroid cholesterylphosphoserine (CPHS) inhibited the secretion of TNF-alpha in lipopolysaccharide-challenged human monocytes. CPHS (5-20 microM) was effective when added together with the endotoxin, or after an interval (1-2 h) sufficient to have allowed for initiation of TNF-alpha synthesis. Consistently, CPHS did not alter TNF-alpha gene transcription. In contrast to its action on TNF-alpha, CPHS showed only marginal effects on interleukin 1beta secretion. Given intraperitoneally to mice 2 h before lipopolysaccharide CPHS prevented the rise in plasma TNF-alpha (IC(50): 5 mg/kg). The inhibition of TNF alpha secretion by CPHS may contribute to the immunosuppressive activity of this steroid. PMID- 10843763 TI - Factors influencing the effect of the soluble IL-6 receptor on IL-6 responses in HepG2 hepatocytes. AB - The soluble IL-6 receptor (sIL-6R) can increase IL-6-induced signalling by forming a complex with IL-6 and membrane-bound gp130 (the receptor beta chain which transduces signals). The conditions affecting this response to sIL-6R were studied using fibrinogen release from HepG2 hepatocytes. Exogenous sIL-6R had no effect alone or in the presence of a submaximal concentration of IL-6, but increased responses to supramaximal IL-6 concentrations in a concentration related manner. Dexamethasone increased the expression of the membrane IL-6R and endogenous sIL6R release, and increased responses to supramaximal but not submaximal IL-6 concentrations. The amount of endogenous sIL-6R released is relatively small and is unlikely to influence the effects of the exogenous sIL 6R. The observed concentration-related decrease in sIL-6R production in the presence of IL-6 may indicate internalization of ligand/receptor complexes. This would significantly decrease the amount of IL-6R (soluble or membrane) available for signalling and limit continued functional response later in the cultures. These data indicate that the major factor influencing responses to exogenous sIL 6R is an excess of IL-6 which is necessary to form complexes with the sIL-6R, which can then interact with gp130 to increase signalling. PMID- 10843764 TI - Synergism between interferon-gamma and cAMP in induction of hepatocyte growth factor in human skin fibroblasts. AB - Interferon (IFN)-gamma stimulates hepatocyte growth factor (HGF) production markedly in various human leukemia cell lines, but its positive effect in human skin fibroblasts is slight. We examined the combined effect of IFN-gamma and various HGF inducers on HGF production in human skin fibroblasts. IFN-gamma synergistically enhanced HGF production stimulated by 8-bromo-cAMP, one of the most effective inducers of HGF: HGF secreted from cells incubated with 1 mM of 8 bromo-cAMP, 1000 U/ml of IFN-gamma and both of these was approximately 8, 1.5 and 24 times, respectively, that secreted from untreated cells. The effect of IFN gamma was dose-dependent and was nullified by an anti-IFN-gamma antibody. Neither IFN-alpha nor IFN-beta had such an enhancing effect, but both these IFNs inhibited the synergistic effect of IFN-gamma and 8-bromo-cAMP. IFN-gamma also synergistically augmented HGF production induced by interleukin-1beta and cAMP increasing agents cholera toxin, forskolin and prostaglandin E(2). HGF gene expression upregulated by cholera toxin, forskolin and 8-bromo-cAMP was markedly enhanced by IFN-gamma, which was detected as early as 3 h after its addition. The synergy between HGF inducers and IFN-gamma is not common to all HGF inducers, because HGF production stimulated by epidermal growth factor and protein-kinase-C activating phorbol esters was significantly inhibited by IFN-gamma. These results indicate that IFN-gamma synergistically stimulates cAMP-induced HGF production and inhibits HGF production induced by growth factors and protein kinase C activators in human skin fibroblasts. PMID- 10843765 TI - Targeting activated lymphocytes with an entirely human immunotoxin analogue: human pancreatic RNase1-human IL-2 fusion. AB - A hybrid human protein was produced in E. coli by fusing the genes encoding human pancreatic RNase1 (hpRNase1) and human IL-2 (hIL-2). The recombinant hpRNase1-hIL 2 inhibited protein synthesis in HTLV-1-infected, malignant T cells, which hyperproduce high affinity IL-2 receptors, with an IC(50)of 2x10(-8) M, whereas no inhibition was detectable in control cells with lower affinity receptors. HpRNase1 alone had an IC(50)of almost 10(-3) M. A molar excess of hIL-2 blocked the protein synthesis inhibition dose-dependently. In a human mixed lymphocyte culture, hpRNase1-hIL-2 inhibited the proliferation of responder cells with potency comparable to that of cyclosporine, while non-effective doses of FK506 importantly improved its potency. Despite its short half-life in animals, hpRNase1-hIL-2 rapidly enters cells in a few minutes and arrests the protein translation in less than 10 h. Thus, hpRNase1-hIL-2 may be useful to selectively eliminate activated lymphocytes hyperproducing high affinity IL-2 receptors, as in allograft rejection, graft-versus-host disease, autoimmune disorders, adult T cell leukaemia and other lymphoproliferative or retroviral malignancies including HIV infection, without inducing general immunosuppression. As an entirely human "immunotoxin analogue" it may alleviate the dose limiting toxicity and immunogenicity of conventional immunotoxins. PMID- 10843766 TI - Metallothionein induction by restraint stress: role of glucocorticoids and IL-6. AB - Restraint stress increased liver metallothionein-I (MT-I) mRNA and MT-I+II protein levels. The glucocorticoid receptor antagonist RU 486 decreased this response. In contrast, adrenalectomy only decreased MT-I+II protein levels. Moreover, corticosterone or progesterone did not reverse the effect of RU 486. These results suggest that glucocorticoids are important for MT-I+II protein synthesis but not for MT-I mRNA accumulation during restraint stress, and that other factors must be involved in this process. Interleukin-6 (IL-6) deficient mice showed a significant decrease of restraint stress-induced liver MT-I mRNA levels (approximately 30% of IL-6+/+ mice) up to approximately 4-5 hours after the onset of stress. Western blotting of hepatic nuclear proteins showed that the IL-6 responsive transcription factor Stat3, which has been shown to mediate MT induction by inflammation, was also activated by restraint stress. Results after extended periods of restraint stress indicate that IL-6 participates early and transiently in the process. The analysis of the expression of the acute phase plasma protein serum amyloid A suggests that restraint stress elicits an acute phase response similar to that caused by inflammation. PMID- 10843767 TI - Effects of interleukin 11 (IL-11) on early post-implantation development of the rat. AB - The development of embryos, trophoblast and decidua of IL-11-treated rats were examined in vivo, while ectoplacental cones (EPC) were studied in vitro. Female Wistar rats were injected daily with buffer (C), 1 mg/kg IL-11 (HD) daily or 30 microgram/kg (LD) IL-11 twice a week. On day 9 of pregnancy, embryonic tissue volume was reduced in IL-11-treated animals, but EPC volume was elevated, compared to controls. Mitotic indices were reduced in embryos (P<0.05 for LD, P<0.001 for HD) and in EPCs of both groups. Pycnotic indices were elevated in LD (NS) and HD (P<0.05) embryos, but decreased in EPCs of the LD group (P<0.01). Morphological abnormalities were observed in decidua, embryo and trophoblast. In HD, EPC attachment was impaired after 1 day culture but proliferation was stimulated after 5 days. Defective decidualization in IL-11 treated rats may therefore result in abnormal development of embryo and trophoblast. PMID- 10843768 TI - TH(1)- and TH(2)-TYPE cytokine expression by activated t lymphocytes from the lung and spleen during the inflammatory response to respiratory syncytial virus. AB - RSV is an important cause of lower respiratory tract illness in infants and the elderly worldwide. The components involved in immunity and those that contribute to inflammation of RSV-induced disease are not clearly understood. To address the relationship between activation antigen and cytokine expression, intracellular levels of IL-2, IL-4, IL-5 and IFN-gamma were determined for CD3, CD44, CD49d, CD54, CD62L and CD102 lymphocytes from the bronchoalveolar lavage and spleen. To examine activation at the DNA level, lymphocytes expressing IL-2, IL-4, IL-5 or IFN-gamma were analysed for G2+M DNA content or phosphatidylserine expression (apoptosis). Trafficking of lymphocytes to the BAL was detected at day 5 p.i., peaked day 7 p.i., and predominately involved CD54(+)and CD102(+)lymphocytes expressing high levels of IL-2, IL-4, IL-5 and IFN-gamma. Lymphocytes expressing CD44(+), CD49d(+)and CD62L(lo)were also observed, however they expressed these cytokines to a lesser extent. DNA analysis of lymphocytes expressing IL-2 or IFN gamma revealed higher G2'M levels compared to lymphocytes expressing IL-4 or IL 5, suggesting greater activation of Th(1)-type lymphocytes in the lung. These data demonstrate that RSV-induced pulmonary inflammation involves extensive cellular activation and cytokine expression, particularly by CD54(+)and CD102(+)lymphocytes in the lung. PMID- 10843769 TI - High inflammation and mild demyelination in the peripheral nervous system induced by an intraneural injection of RR interleukin-4. AB - To examine the direct effects of IL-4 on peripheral nervous system (PNS) we injected recombinant rat IL-4 (rrIL-4) into the sciatic nerve of normal adult Lewis rats. Histopathological and immunohistochemical observations revealed that 1 day after injection, a large number of macrophages and MHC class II-positive cells appeared within both the perineurium and endoneurium. Only few CD4(+)and CD8(+)T cells existed in the endoneurium. From day 4 to day 7, we observed a gradual decline of inflammation, but the number of infiltrates in rrIL-4 injected nerves was significantly higher compared with sterile PBS-injected control group. On the contrary, demyelination affected significantly fewer nerve fibres in the rrIL-4-injected nerves compared with control group on day 7. Intraneural injection of rrIL-4 results in high grade inflammation and mild demyelination in the PNS. PMID- 10843770 TI - Transforming growth factor beta1 and soluble Fas serum levels in hepatocellular carcinoma. AB - In this study we assessed the usefulness of serum Transforming Growth Factor beta1 (TGF-beta1) and soluble Fas (sFas) in distinguishing liver cirrhosis (LC) with and without hepatocellular carcinoma (HCC) as compared with alpha fetoprotein (AFP). Serum TGF-beta1 and sFas levels were measured by ELISA in 51 LC patients, 54 patients with HCC and 30 healthy donors. Considering as a cut-off limit (mean+1SD of controls) 74 pg/ml and 637 pg/ml for TGF-beta1 and sFas, respectively, we computed serum concentrations of TGF-beta1 and sFas as a score (mean+/-SD). The positive frequency of serum TGF-beta1 levels in HCC patients (54%) was greater than in LC patients (26%) and healthy donors (3%). TGF-beta1 levels were higher in HCC (1.6+/-0.5) than in LC (1.1+/-0.2) (P<0.0001) and healthy donors (0.6+/-0.2). Using a cut-off limit of 82 pg/ml (mean+2SD), the positive frequency of TGF-beta1 was 20% in HCC patients. None of the controls and LC patients had TGF-beta1 levels higher than 82 pg/ml. The positive frequency of serum sFas levels was 100% in HCC patients, 98% in LC patients and 3% in healthy controls. Serum sFas levels were higher in HCC (2.5+/-0.7) than in LC (1.9+/-0.5) (P<0. 001) and healthy donors (0.6+/-0.3). No significant change of positive frequency was obtained by setting sFas cut-off at higher levels. sFas values did not correlate with TGF-beta1 levels. No relationship was found between TGF-beta1 amounts and AFP levels. However, in the 23% of HCC patients, with normal AFP values TGF-beta1 levels were higher than the cut off. These findings suggest the potential usefulness for TGF-beta1 assay in AFP-negative HCC. PMID- 10843771 TI - Soluble CD30 serum level in HCV-positive chronic active hepatitis: A surrogate marker of disease activity? AB - In the present study, high levels of CD30s, a glycoprotein preferentially expressed and released by T lymphocytes producing Th(2)-type cytokines, were seen in the sera of patients with chronic hepatitis C, and a correlation with histological activity of the disease was found. CD30s levels were assayed in the sera of 29 HCV RNA-positive patients with histologically proven chronic active hepatitis and in 30 healthy blood donors. Thirteen of 29 (45%) HCV patients had CD30s serum levels above the normal range (>20 U/ml). Mean CD30s serum levels were significantly higher in HCV patients than in controls (P<0.0005). A positive correlation was found between serum CD30s levels and both the histological activity index (r=0.59, P=0.001) and ALT serum levels (r=0.5; P=0.006). The raised CD30s level found in more severe HCV liver disease indirectly suggests activation and expansion of Th(2)cells. CD30s levels could represent a useful surrogate marker of activity in chronic HCV infections. PMID- 10843772 TI - Differences in the genotypes and plasma concentrations of the INTERLEUKIN-1 receptor antagonist in black and white South African asthmatics and control subjects. AB - The allelic frequency of a variable tandem repeat (VNTR) polymorphism in intron 2 of the IL-1 Ra gene was studied in black and white patients with asthma as well as control individuals. The plasma IL-1 Ra concentration was also determined in asthmatics and compared to control individuals. The 410-bp allele of the IL-1 Ra was significantly increased in all black subjects (90%) as compared to all white subjects (74%, P<0.0001), while the 240-bp allele was significantly reduced in all black subjects (11%) as compared to all white subjects (27%, P<0.0001). There was no difference in the frequency of the VNTR of the IL-1 Ra between black asthmatics and black controls and between white asthmatics and white controls. The IL-1 Ra levels were significantly increased in black and white patients with severe or moderate asthma as compared to patients with mild asthma. Increased plasma concentrations of the IL-1 Ra was found to be associated with disease severity in all asthmatic patients. PMID- 10843773 TI - Inflammatory and immunological parameters in children with haemolytic uremic syndrome (HUS) and gastroenteritis-pathophysiological and diagnostic clues. AB - The objective of this study was to identify parameters indicating a risk for developing typical haemolytic uremic syndrome (D+HUS) during the prodromal phase of diarrhea caused by enterohaemorrhagic Escherichia coli (EHEC). Forty-eight children were studied prospectively with regard to inflammatory serum factors on admission to hospital. Ten patients developed D+HUS (group I), 15 suffered from viral-gastroenteritis (group IIa) and 23 from other types of bacterial gastroenteritis (group IIb). Mean levels of IL-8 tended to be elevated in group I compared to groups IIa and IIb. Neopterin and IL-10 levels particularly were significantly decreased in HUS in comparison to both gastroenteritis groups. Low IL-10 levels indicate a substantial disregulation of the immune response in HUS, as IL-10 downregulates the pro-inflammatory response and suppresses pro-coagulant activity in experimental endotoxemia. Our results suggest low neopterin, high IL 8 and especially low IL-10 levels are indicators of a high risk for developing HUS. PMID- 10843774 TI - Comparison of non-viral transfection methods in melanoma cell primary cultures. AB - Melanoma primary cultures were transiently transfected via electroporation and lipofection for comparison. Transfection efficiency was superior with electroporation (58+/-9%) as compared to lipofection (23+/-9%) as determined by enhanced green fluorescent plasmid (EGFP) transfection. Secretion of IL-2 persisted for up to 3 weeks after electroporation. The increase in sensitivity against immunologic effector cells by transfection with IL-2 was not significant. Our results show the feasibility of a gene transfer into primary human melanoma cells, different from retroviral transduction. PMID- 10843775 TI - The spatial and temporal expression patterns of netrin receptors, DCC and neogenin, in the developing mouse retina. AB - Recently it has been demonstrated that the guidance of retinal ganglion cell (rgc) axons through the optic disc is dependent on the DCC/netrin-1 axonal guidance system. To gain further insight into the function of the netrin receptors, DCC and Neogenin, in retinal development we have studied the expression patterns of these receptors in the embryonic mouse retina. Neogenin mRNA was restricted to a single neural cell type, the rgc. However, strong Neogenin mRNA expression was observed in the extending fiber cells of the developing lens suggesting a role for Neogenin in the migration events shaping the early lens. Our studies demonstrated that DCC mRNA was expressed at high levels in chains of closely opposed neurons as they migrated towards the emerging mantle layer in the early retina (E12.5-E13.5) suggesting a role for DCC in the migration of neurons out of the ventricular zone. DCC protein expression was high on rgc axons as they actively navigated through the optic disc into the optic nerve. At birth, when the majority of rgc axons had projected through the optic disc, DCC protein was no longer detectable on the distal axonal segments within the optic nerve despite significant DCC protein expression on the proximal axonal membranes in the nerve fiber layer. These observations suggest that a localized down-regulation of DCC protein occurs on projecting axonal membranes once the DCC guidance function is no longer required. We also demonstrated that DCC mRNA and protein were expressed by amacrine cells and Muller glial cells while DCC mRNA was detected in horizontal cells. Taken together, these expression patterns suggest a role for DCC in axon outgrowth and/or pathfinding for a variety of retinal neurons and in the migration of newly born neurons within the developing retina. PMID- 10843776 TI - Diabetes reduces glutamate oxidation and glutamine synthesis in the retina. The Penn State Retina Research Group. AB - Retinas of diabetic individuals develop early functional changes measurable by electrophysiological and psychometric testing. Using a rat model of diabetes, we previously identified diabetes-induced alterations in metabolism of the neurotransmitter glutamate which may ultimately lead to accumulation of glutamate in the retina (Diabetes, 47: 815, 1998). We therefore investigated the function of enzymes that mediate the synthesis and breakdown of glutamate in retinas from rats made diabetic by injection of streptozotocin. De novo synthesis of nitrogen containing amino acids including glutamate, glutamine and aspartate was assessed by measuring the rate of carbon fixation in freshly dissected retinas, and was unchanged by diabetes. In contrast, the oxidation of glutamate was significantly reduced in retinas from diabetic rats (62%, P < 0.05). Furthermore, diabetic retinas were less susceptible to inhibition of glutamate oxidation by the transaminase inhibitor aminoxyacetate (80%, N.S.), compared to the significant decrease seen in control rats (61%, P < 0.001). The activity and content of glutamine synthetase were also significantly reduced in retinas from rats diabetic for 2-6 months [range of 48% (P < 0.005) to 83% (P < 0.05) compared to control]. The activity of glutamine synthetase was normalized by acute injections of insulin, but not by reducing blood sugar levels with injections of phlorizin. These results indicate two enzymatic abnormalities in the glutamate metabolism pathway in the retina during diabetes: transamination to alpha-ketoglutarate and amination to glutamine. The reduced flux through these pathways may be associated with the accumulation of glutamate. These results are also consistent with the possibility that some of the glial changes in the retina during diabetes may be caused by hypoinsulinemia rather than hyperglycemia. PMID- 10843777 TI - Xenopus IRBP, a phylogenetically remote protein, is uveitogenic in Lewis rats. AB - Mammalian interphotoreceptor retinoid-binding proteins (IRBPs) are highly uveitogenic in Lewis rats. Xenopus laevis IRBP resembles mammalian IRBP in its four-fold structure, and has approximately 70% amino acid sequence identity with the bovine protein. This study investigated the uveitogenicity of recombinant Xenopus IRBP and two of its derived peptides in Lewis rats. Rats immunized with Xenopus IRBP developed uveoretinitis as well as pineal inflammation. The Xenopus molecule was, however, less immunopathogenic than the bovine IRBP. Of the two Xenopus IRBP peptides tested, 1180-1191 was remarkably uveitogenic, whereas sequence 521-540 exhibited low activity. It is assumed, therefore, that as with bovine IRBP, peptide 1180-1191 is the major uveitogenic sequence in Xenopus IRBP. The role individual residues of these peptides play in the immunopathogenic process is discussed. Our data thus demonstrate that despite its being phylogenetically remote, Xenopus IRBP is uveitogenic in Lewis rats PMID- 10843778 TI - Preservation of donor cornea prevents corneal allograft rejection by inhibiting induction of alloimmunity. AB - To determine whether preservation of the donor cornea prevents allograft rejection, orthotopic corneal transplantation was performed using corneas preserved in storage medium (Optisol-GS((R))). Donor corneas harvested from C3H/He (H-2(k)) mice and B10.D2 (H-2(d)) mice were preserved in storage medium for 0, 3, 7 and 14 days, and then transplanted into the corneal beds of the recipient BALB/c (H-2(d)) mice. Graft survival was determined clinically and histologically. The expression of major histocompatibility complex (MHC) molecules in the preserved corneas was analysed by immunohistochemistry and Western blotting. Donor-specific cytotoxic T lymphocyte (CTL) and delayed-type hypersensitivity (DTH) responses were assessed 3 weeks after grafting. Active suppression of DTH in the recipient mice was also examined 3 weeks after grafting. The survival of 14 day preserved allografts was significantly higher than that of the non-preserved allografts in both MHC and minor histocompatibility (H) antigens, and minor H only disparate combination. The recipients of the preserved allografts failed to induce both CTL and DTH. The active suppression of DTH was not acquired in these recipients. The expression of donor-derived MHC class I antigens was markedly reduced in the corneas after preservation. Preservation of the donor cornea had a remarkable effect on the prevention of corneal allograft rejection. Since the preserved allografts failed to induce donor-specific CTL and DTH, and active suppression of DTH was not acquired in the recipients, the prevention of allo-rejection is due to a failure of allo-sensitization. These results indicate that the reduction of MHC class I antigens and minor H antigens expression in the preserved grafts induces a failure of allo-sensitization and leads to the high rate of acceptance in corneal allografts. PMID- 10843779 TI - Modulation of lens glycolytic pathway by thioltransferase. AB - The observation that the level of S-thiolated proteins (protein-thiol mixed disulfides) was transiently increased in the lens epithelial cells correlation with the transient inactivation of glyceraldehyde-3-phosphate dehydrogenase (G 3PD), a key glycolytic enzyme, when the cells were treated with a bolus of hydrogen peroxide, prompted our speculation that G-3PD may have been transiently thiolated at the SH sensitive active center. In the meantime, thioltransferase (TTase), a thiol regulating enzyme, whose activity remained constant under the same condition, may be regulating G-3PD and other sulfhydryl-sensitive glycolytic enzymes through thiol-disulfide exchange reactions ( Lou et al., 1998 ). To prove this hypothesis, several purified glycolytic enzymes from a commercial source, including hexokinase (HK), G-3PD, pyruvate kinase (PK) and fructose 1,6 bisphosphatase (FBPase), an enzyme in gluconeogenesis, were made into protein thiol mixed disulfide and used for this study. Glycolytic enzymes in cultured rabbit lens epithelial cells pre-exposed to H(2)O(2)(0.5 m M for 15 min) were also studied for this purpose. Recombinant human lens thioltransferase (RHLT), which was isolated and purified previously in this laboratory, reactivated these pure glycolytic enzymes inactivated by forming protein-S-S-gluthathione (PSSG), protein-S-S-cysteine (PSSC) or, protein-S-S-cysteamine after thiolating with oxidized glutathione, cystine or cystamine respectively. RHLT also reactivated these enzymes in the cell extract of cultured rabbit lens epithelial cells after being briefly exposed to 0.5 m M H(2)O(2). The S-thiolation and dethiolation of FBPase however, showed an opposite effect to that of glycolytic enzymes. These results suggest that TTase may participate in the repair process of glycolytic enzymes during oxidative stress and restore their activities in situ. PMID- 10843781 TI - The human hyaloid system: cell death and vascular regression. AB - The present study had investigated the roles of apoptosis and necrosis in the regression of the human fetal hyaloid vasculature. Normal human fetal hyaloid specimens (n = 67) ranging from 10 to 20 weeks' gestation were studied. Specimens were either immunolabeled with anti-von Willebrand factor and major histocompatibility complex class I antibodies or investigated using the terminal deoxyribonucleotidyl transferase-mediated dUTP-biotin DNA nick-end labeling technique. A fluorescent DNA-binding dye acridine orange/ethidium bromide mixture was also applied to unfixed flat mounts of hyaloid vasculature and some specimens were processed for transmission electron microscopy. Vascular regression including cell loss in the connecting vessels, stretching and thinning of the vasa hyaloidea propria, tunica vasculosa lentis and the pupillary membrane was clearly evident after 13 weeks' gestation. Cresyl violet staining revealed condensed cells and pyknotic bodies throughout the hyaloid system; cell death occurred either in single cells or along small capillary segments associated with vascular regression. Acridine orange/ethidium bromide staining showed DNA condensation at early and late stages of cell death. Similarly, DNA nick-end labeling was positive in endothelial cells, pericytes and vessel and non-vessel associated hyalocytes. The observation of hyalocytes juxtaposed to cytolysed endothelial cells may indicate a role for these cells in vascular regression. Features of apoptosis were more evident during early vascular regression whilst necrosis was increasingly evident at later stages. PMID- 10843780 TI - Signal transduction mechanisms involved in ischemic preconditioning in the rat retina in vivo. AB - Ischemic preconditioning (IPC) protects the rat retina against the injury that ordinarily follows severe ischemia. We showed previously that release of adenosine and de novo protein synthesis were required for IPC protection. The mechanisms of IPC were studied in the rat retina by examining the signal transduction mediators responsible, in particular, those theorized to be downstream of adenosine receptors. In addition, we examined the hypothesis that nitric oxide and hydroxyl radicals were involved in the IPC protective phenomenon. Retinal ischemia was produced for 60 min in ketamine/xylazine anesthetized Sprague-Dawley rats, and recovery was measured using electroretinography. We tested the effects on the protective effect of IPC resulting from antagonism of protein kinase C, potassium ATP channels, nitric oxide synthase, or hydroxyl radicals. The effects of the inhibition of de novo protein synthesis or of protein kinase C, and blockade of potassium ATP channels on the mimicking of IPC by adenosine receptor agonists was examined.IPC protection was strongly attenuated by inhibition of protein kinase C and by blockade of potassium ATP channels, but unaffected by the inhibition of hydroxyl radicals. Blockade of nitric oxide synthase produced a trend toward enhancement of IPC protection. Mimicking of IPC protection by adenosine receptor agonists was inhibited by blockade of protein synthesis or of protein kinase C, as well as by potassium ATP channel antagonism. These results demonstrate that protein kinase C and potassium ATP channels are mediators of the protective effect produced by IPC. In addition, the results show that stimulation of adenosine receptor subtypes A1 and A2a is responsible for IPC protection via downstream stimulation of protein kinase C, the opening of potassium ATP channels, and de novo protein synthesis. PMID- 10843782 TI - Corneal insult affects the production and distribution of FGF-2 within the lacrimal gland. AB - The purpose of this study was to determine the distribution of FGF-2 within rabbit lacrimal glands and to determine whether corneal insult affects that distribution. The scarified corneas of experimental animals were inoculated either with adenovirus type 5 or buffer. Control animals were either untreated, or animals whose corneas were scarified. Twenty-one days later all animals were killed and the lacrimal glands were studied by immunocytochemistry and Western blotting to detect FGF-2. In untreated control animals, FGF-2 was immunolocalized predominantly within a population of elongated cells in the basal epithelium of ducts, and to a lesser degree in the basal epithelium of the acini. The elongated immunopositive cells appear to be myoepithelial cells known to be present at these sites. Interstitial cells around ducts and acini, and the basement membranes of the ducts and acini, were also immunopositive for FGF-2. Twenty-one days after adenovirus inoculation and scarification of the cornea, immunopositivity for FGF-2 was dramatically decreased in basement membranes, but increased within myoepithelial cells of the duct epithelium. These myoepithelial cells were frequently enlarged, bulging toward the duct lumen. In animals whose corneas were inoculated with buffer and scarified, or animals whose corneas were simply scarified, the changes in the lacrimal gland were similar, but somewhat less pronounced, to those of adenovirus-inoculated animals. Western blots confirmed the presence of FGF-2 immunoreactivity in all groups. The major band in untreated controls was at 24 kD, whereas all animals with corneal scarification had major bands at 38 kD. Densitometry of Western blots demonstrated that the amount of 24 kD FGF-2 present within the lacrimal gland after corneal scarification was at least 50% less than in untreated controls, whereas 38 kD FGF 2 was at least ten-fold greater. Our findings indicate that corneal scarification results in an altered distribution of FGF-2 within the lacrimal gland, which involves a decrease in low molecular weight FGF-2 and a dramatic increase in a higher molecular weight isoform of FGF-2. FGF-2 may be released from myoepithelial cells apically (exocrine) into the tear fluid and basally (autocrine/paracrine) into the connective tissue, as well as from extracellular complexes within basal laminae. PMID- 10843783 TI - Insulin and IGF-I affect the protein composition of the lens fibre cell with possible consequences for cataract. AB - Explanted newborn rat lens epithelial cells were cultured with various concentrations of FGF-2 and/or insulin or IGF-I for 8-20 days. The accumulation of alphaA-, alphaB-, betaA3/1-, betaB2- and gammaA-F-crystallin was measured. During culture with insulin only, i.e. in the absence of fibre cell differentiation, alphaA- and alphaB-crystallin accumulated to the same level as found in differentiating cells. Culture of epithelial cells with IGF-I led to an increase in alphaB-crystallin, but not in alphaA-crystallin. The addition of insulin under differentiation conditions (in the presence of 25 ng ml(-1)FGF-2) augmented the accumulation of alphaA-crystallin 1.5-fold, the accumulation of betaB2-crystallin two-fold and the accumulation of gammaA-F-crystallin five-fold over that found with FGF-2 only. The accumulation of alphaB- and betaA3/1 crystallin was not affected by insulin in the presence of FGF-2. Adding IGF-I to fibre cells differentiating in the presence of 25 ng ml(-1)FGF-2 resulted in a 1.5-fold increase (of questionable statistical significance) in both alphaA- and alphaB-crystallin and a two to three-fold increase in gammaA-F-crystallin compared to cells cultured with FGF-2 only, no significant effect of IGF-I on the accumulation of betaA3/1- or betaB2-crystallin was found. Comparison of the levels of mRNA and protein suggests that insulin acts to increase the level of transcription. Our results show that the response of fibre cells to insulin or IGF-I differs. Hence, even though half the maximum dosage required for the insulin effect was rather high (between 0.1 and >5 micro g), the effect of insulin cannot be merely transmitted by the IGF-I receptor. Our data further predict that insulin or IGF-I increases the overall ratio of beta- and gamma crystallin to alpha-crystallin in the fibre cell, which could predispose the lens to cataract. PMID- 10843784 TI - Thiol disulfide exchange modulates the activity of aldose reductase in intact bovine lens as a response to oxidative stress. AB - The reversibility of S-thiolation of aldose reductase was shown in intact bovine lens subjected to oxidative stress. The glutathione modified aldose reductase generated in the lens as a consequence of hyperbaric oxygen treatment was recovered in its reduced form following culturing in normobaric air conditions. Nucleus and cortex were differently affected by both oxidative treatment and normobaric air recovery. The extent of S-thiolation of aldose reductase appeared to be higher in the nucleus than in the cortex. Moreover, the nucleus, but not the cortex, was unable to completely recover from the protein S-thiolation process. The ratios of GSH/GSSG and NADPH/NADP(+)as well as the Energy Charge values were determined in the cortex and nucleus both after oxidative stress and recovery. The results are consistent with the existence of a quite well-defined boundary between the two lens regions. Moreover, they are supportive of the hypothesis that thiol/disulfide exchange has the potential to be a regulatory mechanism for certain enzymes which can modulate the flux of NADPH inside the cell. PMID- 10843785 TI - Circadian rhythm of serotonin N -acetyltransferase activity in rat lens. AB - Activity of serotonin N -acetyltransferase (NAT), a key regulatory enzyme in melatonin biosynthesis, was detected in the rat lens. NAT activity in the lens showed significant diurnal variation in vivo and in vitro, peaking during the period of darkness, when the lenses were maintained under 14 hr light/10 hr dark cycle. Cultured lenses exhibited a circadian rhythm of NAT activity when maintained under constant darkness. However, the rhythm in vitro was not entrained when the light/dark cycle was delayed 8 hr from the cycle in intact animals. These data strongly suggest that the rat lens contains a circadian clock that controls NAT activity, although the circadian clock appears to lack a photic entrainment mechanism. PMID- 10843786 TI - Transducin binding in bovine rod outer segment disk membranes of different age/spatial location. PMID- 10843788 TI - Cloning of the cDNA for sea bream (Sparus aurata) parathyroid hormone-related protein. AB - This paper reports cloning of the cDNA for sea bream (Sparus aurata) parathyroid hormone-related protein (PTHrP). The gene codes for a 125-amino acid mature protein with a 35-residue prepeptide. The total gene sequence is 1.8 kb with approximately 75% noncoding. The N-terminus of the protein resembles mammalian and chicken PTHrP peptides with 12 of the first 21 amino acids identical and for which there is homology with mammalian parathyroid hormone. Toward the C terminus, the nuclear transporter region between residues 79 and 93 in sea bream is 73% homologous to tetrapod PTHrP, and the RNA binding domain, 96-117, is 50% homologous, moreover starting with the conserved lysine and terminating with the lysine/arginine sequence. Sea bream PTHrP differs significantly from mammalian and chicken PTHrP, having a novel 16-amino acid segment between residues 38 and 54 and completely lacking the terminal domain associated in mammals with inhibition of bone matrix lysis. RT-PCR and in situ hybridization of sea bream tissues show that the gene is expressed widely and the results confirm observations of a PTHrP-like factor in sea bream detected with antisera to human PTHrP. PMID- 10843787 TI - The effects of highly selective opioid receptor antagonists on the release of arginine vasotocin induced by hyperosmotic stimulation and angiotensin II injection. AB - The effects of highly selective antagonists to mu-, delta-, and kappa-opioid receptor subtypes on hyperosmotic- or angiotensin II (AII)-induced arginine vasotocin (AVT) release were investigated in chicks. Plasma levels of AVT increased about 1.5-fold after the administration of 1.5 M NaCl (200 microl, ip) or 100 ng AII (5 microl, icv). The administration of the mu-antagonist naloxonazine and the kappa-antagonist nor-Binaltorphimine further elevated plasma levels of AVT stimulated by hypertonic NaCl or AII. These effects of mu- and kappa-opioid receptor antagonists on AVT release were dose dependent. Nor Binaltorphimine enhanced hyperosmotically stimulated plasma levels of AVT at a lower dose than that of naloxonazine. Conversely, the delta-selective antagonist naltrindole did not significantly affect AVT secretion. None of the opioid receptor antagonists influenced basal plasma levels of AVT. Therefore, these results suggest that mu- and kappa-opioid receptors are involved in hyperosmotic- and AII-induced AVT release, and the effect of the kappa-opioid receptor antagonist in the AVT release stimulated by hyperosmolality is strong compared to that of the mu-opioid receptor antagonist. PMID- 10843789 TI - Oocyte plasma membrane proteins and the appearance of vitellogenin binding protein during oocyte growth in the lizard Podarcis sicula. AB - In the ovary of the lizard Podarcis sicula, the micropinocytotic uptake of the yolk exogenous precursor (i.e., vitellogenin; VTG) occurs only in the reproductive period and involves the plasma membrane of > or =2000-microm oocytes. This paper analyzes the intrinsic proteins extracted from the plasma membrane of growing oocytes to identify the vitellogenin binding protein during the different stages of the annual ovarian cycle of this species. Despite the well-known ultrastructural changes of the oocyte plasma membrane, SDS-PAGE failed to show marked variation in the total number of membrane proteins during the most significant stages of oocyte auxocytosis. Nevertheless, ligand blotting, using homologous VTG and anti-VTG, revealed that an congruent with115-kDa protein of the oocyte plasma membrane bound plasma vitellogenin only in the reproductive period (spring-summer) in both vitellogenic and nonvitellogenic oocytes. During the nonreproductive period, this molecule was never observed. However, it could be induced in the coldest months (winter) by hypophyseal gonadotropins. PMID- 10843790 TI - Glucose stimulates the release of bombyxin, an insulin-related peptide of the silkworm Bombyx mori. AB - The effects of starvation and feeding on the release of bombyxin, a peptide of insulin superfamily in insects, from the larval brain of the silkworm Bombyx mori were investigated. Following starvation, the bombyxin titer in the hemolymph of larvae decreased, whereas its content in the brain increased. On the other hand, refeeding of the starved larvae resulted in an increase in the hemolymph bombyxin titer and a rapid decrease in the hormone level in the brain. These results indicate that the release of bombyxin from the brain is suppressed by starvation and stimulated by feeding. The hemolymph glucose titer also changed sharply upon starvation and refeeding, and a close relationship was observed between the changes in glucose concentrations and bombyxin titers in the hemolymph. The injection of glucose into starved larvae could mimic the effect of refeeding on the release of bombyxin, suggesting that glucose serves as the signal for the "fed" state of the animal. It is likely that glucose is a common nutritional signal for inducing the release of mammalian and insect insulins. PMID- 10843791 TI - Short-chain fatty acids inhibit the release and content of growth hormone in anterior pituitary cells of the goat. AB - The effects of short-chain fatty acids (SCFA: acetate, propionate, and butyrate) on growth hormone (GH)-releasing hormone (GHRH)-induced GH secretion from pituitary somatotrophs were assessed on isolated anterior pituitary cells of goats. Cells were cultured in Dulbecco's modified Eagle's medium for 3 days, either in the presence (1, 3, or 10 mM) or in the absence of each SCFA, and then stimulated with GHRH (10(-12) to 10(-7) M) for 30 min, again in the presence of and at the concentration of SCFA used over the previous 3 days. In the cells cultured in the absence of SCFA, the addition of SCFA to the medium during the GHRH stimulation period did not significantly change GHRH-induced GH release. However, in cells cultured in the presence of either propionate (3 or 10 mM) or butyrate (1, 3, or 10 mM), the addition of SCFA to the medium during GHRH stimulation significantly reduced the GHRH-induced GH release. The inhibitory effects of SCFA were dependent on the concentrations of SCFA and were greater for butyrate than for propionate. In the cells cultured in the presence of butyrate, but not in the absence, the total GH production (the sum of the released GH and the remaining GH after stimulation) was also significantly reduced. The GHmRNA expression was reduced in the cells cultured with 10 mM butyrate, whereas it was enhanced by the stimulation with 10(-7) M GHRH. These findings suggest that propionate and butyrate may inhibit GHRH-induced GH release and GH production by caprine anterior pituitary cells. PMID- 10843792 TI - Evidence for a hormonal tactic maximizing green turtle reproduction in response to a pervasive ecological stressor. AB - Mortality of breeding sea turtles due to excessive heat exposure after nesting activities is an unusual feature of the Raine Island green turtle rookery. Breeding turtles that fail to return to the ocean after oviposition can experience increasing body temperatures that exceed lethal limits (>39 degrees C) as ambient temperatures rise after sunrise. We investigated how acute increases in body temperature influenced plasma corticosterone (B) concentrations of individual turtles. Furthermore, interactions between progesterone (P) and testosterone (T) and increasing body temperature and the glucocorticoid corticosterone were examined for negative correlations. Breeding green turtles exhibited a 16-fold mean increase in plasma corticosterone concentration as body temperature (cloacal) rose from 28.2 to 40.7 degrees C in less than 6 h. However, the absolute increase in plasma B was small and much less than expected, despite the lethal stressor. Comparatively, the maximal B response to lethal heat stress was similar to plasma B concentrations obtained from breeding female turtles exposed to 8 h of capture stress. However, the maximal B response of breeding turtles exposed to heat and capture stressors was significantly less than the B response of nonbreeding adult female turtles subjected to an 8-h capture stressor. No negative correlations were observed between plasma T and plasma B, between plasma T and body temperature, between plasma P and plasma B, or between plasma P and body temperature. Our findings provide further evidence that reduced adrenocortical function operates in breeding green turtles in the presence of even the most pervasive of environmental stressors. PMID- 10843793 TI - Effects of gonadotropin-releasing hormone analog on expression of genes encoding the growth hormone/prolactin/somatolactin family and a pituitary-specific transcription factor in the pituitaries of prespawning sockeye salmon. AB - Gonadotropin-releasing hormone (GnRH) is a possible secretagogue of growth hormone (GH) and somatolactin (SL) in teleosts. Effects of GnRH on the levels of pituitary mRNAs encoding GH, prolactin (PRL), and SL were therefore examined in prespawning sockeye salmon (Oncorhynchus nerka). A capsule of GnRH analog (GnRHa) was implanted into the dorsal muscle of maturing sockeye salmon for 3 weeks. The levels of hormonal mRNAs were then determined by a quantitative dot blot analysis using single-stranded sense DNA of the same sequence of mRNA as the standard. Further, we analyzed effects of GnRHa on expression of the genes encoding pituitary-specific transcription factor (Pit-1/GHF-1). Relative levels of Pit 1/GHF-1 mRNAs were estimated by Northern blot analysis, which showed specific 2- and 3-kb bands of mRNAs. GnRHa significantly increased the level of SL mRNA in the males, but not in the females, compared to the control fish. It did not induce significant increases in the levels of GH and PRL mRNAs in both the males and the females. The levels of Pit-1/GHF-1 mRNAs in the control males tended to be higher than those in the initial controls, so that GnRHa might not be effective in enhancing expression of Pit-1/GHF-1 gene, except for the level of 3 kb Pit-1/GHF-1 mRNA in the females treated with 150 microg GnRHa. The pattern of changes in the levels of Pit-1/GHF-1 mRNAs were similar to those of GH and PRL mRNAs in both the males and the females and to that of SL mRNA in the females. These results indicate that, in prespawning sockeye salmon, GnRH can stimulate SL gene expression, but probably not through the Pit-1/GHF-1-dependent system. PMID- 10843794 TI - Year-class differences in the reproductive system, plasma prolactin and corticosterone concentrations, and onset of prebasic molt in male dark-eyed juncos (Junco hyemalis) during the breeding period. AB - Year-class differences in reproductive function were investigated in a free living population of adult male Dark-eyed Juncos, Junco hyemalis, breeding in interior Alaska. Second-year males (SY, entering their first breeding season) were compared with after-second-year males (ASY, entering at least their second breeding season). We measured body mass, size of the cloacal protuberance (CP), testis mass, onset of prebasic molt, and concentrations of plasma luteinizing hormone (LH), testosterone (T), corticosterone (CORT), and prolactin (PRL) throughout the reproductive season (April to mid-July). There were no differences in SY and ASY body weights but SY males had smaller CPs and testis masses than ASY males during gonadal recrudescence and at the end of the breeding season. Plasma LH was elevated from April until mid-June and then decreased in the same way in both year classes. In contrast, plasma T was high from April until mid-May and was lower in SY than in ASY juncos shortly after they arrived on their breeding grounds at the end of April, but not at other times. In July, SY males started to molt earlier, suggesting that they became photorefractory earlier than ASY males. Plasma PRL increased progressively in both year classes between April and early June and decreased in early July. At this time, plasma PRL decreased earlier in SY than in ASY males. Plasma CORT changed seasonally, but did not differ between SY and ASY juncos. Thus, year-class differences in CP sizes and testis mass apparently did not result from SY males secreting less LH or more PRL or CORT than ASY males. It is suggested that differences in reproductive condition in SY and ASY juncos are mediated by interactions with conspecific birds and do not result from an intrinsic effect of age. PMID- 10843795 TI - Effects of salmon gonadotropin-releasing hormone on follicle stimulating hormone secretion and subunit gene expression in coho salmon (Oncorhynchus kisutch). AB - Previous work has indicated that, during the process of gametogenesis in salmon, follicle-stimulating hormone (FSH) and luteinizing hormone (LH) are differentially synthesized and released. Although substantial information is available on the regulation of LH in many fish species, relatively little is known about the regulation of FSH biosynthesis and secretion or the regulation of two types of alpha subunit in salmon. In this study, the effects of salmon gonadotropin-releasing hormone (sGnRH) on in vitro secretion of FSH, and alpha1, alpha2, LH beta, and FSH beta subunit gene expression were investigated in coho salmon (Oncorhynchus kisutch) using primary pituitary cell cultures. To quantify FSH beta, LH beta, alpha1, and alpha2 subunit transcript levels, a multiplex RNase protection assay (RPA) was developed. Probes for the beta subunits of coho salmon FSH and LH were available from previous studies. To generate probes for the alpha subunit RPAs, alpha1 and alpha2 subunit cDNAs were cloned using reverse transcriptase PCR. Release of FSH and LH into cell culture medium was quantified by radioimmunoassays. The effects of sGnRH on gonadotropin release and gene expression were tested at two points during the spring (April and May) prior to spawning in the autumn; a period when plasma and pituitary FSH levels are increasing and females are in early stages of secondary oocyte growth. In both experiments, sGnRH increased steady-state mRNA levels of FSH beta, alpha1, and alpha2, whereas LH beta mRNA levels were not detectable. Secretion of FSH was stimulated by sGnRH in a concentration-dependent manner. Medium LH was not detectable in the first experiment (April) and was measurable only after sGnRH treatment in the second experiment (May). Control levels of medium FSH and transcripts for FSH beta and alpha1 subunits increased approximately fourfold between April and May, whereas alpha2 transcript levels remained relatively constant, suggesting that the seasonal increase in FSH release may involve increased production of alpha1. Therefore, sGnRH has direct stimulatory effects on both secretion of FSH and FSH subunit biosynthesis, most likely due to increased transcription. However, alterations in rates of transcript degradation cannot be ruled out. PMID- 10843796 TI - Intermale competition in sexually mature arctic charr: effects on brain monoamines, endocrine stress responses, sex hormone levels, and behavior. AB - Sexually mature Arctic charr (Salvelinus alpinus) males were allowed to interact in pairs for 4 days in the absence of females. Agonistic behavior was quantified, and at the end of the experiment, plasma levels of glucose, cortisol, testosterone (T), 11-ketotestosterone (11-KT), and 17alpha,20beta-dihydroxy-4 pregnen-3-one (17alpha, 20beta-P) were determined alongside brain concentrations of serotonin (5-HT), 5-hydroxyindoleacetic acid (5-HIAA, the major 5-HT metabolite), dopamine (DA), and 3,4-dihydroxyphenylacetic acid (DOPAC, a major DA metabolite). Plasma cortisol and glucose were significantly elevated in subordinate fish, and the number of aggressive acts received showed positive correlations with plasma levels of glucose on day 1, during the development of the dominance relationship, and of cortisol on day 4, when the dominance relationship was established. In contrast, plasma concentrations of T and 11-KT were significantly higher in dominant than in subordinate males, and there was a similar tendency in plasma concentrations of 17alpha,20beta-P. Further, plasma levels of these gonadal steroids were correlated with the number of aggressive acts performed on day 4, but not with the number of aggressive acts received. The plasma cortisol concentrations did not correlate with either 5-HIAA:5-HT or DOPAC:DA ratios in any of the brain parts analyzed. Plasma glucose levels showed positive correlations with brain 5-HIAA:5-HT ratios. Negative correlations were observed between 5-HIAA:5-HT ratios in the optic tectum and between plasma levels of T, 11-KT, and 17alpha,20beta-P. Telencephalic DOPAC/DA ratios displayed a negative correlation with plasma levels of T, 11-KT, and 17alpha,20beta-P, but only in dominant males. PMID- 10843797 TI - Evidence that nitric oxide is involved in the regulation of growth hormone secretion in goldfish. AB - Whether nitric oxide (NO) plays a role in regulation of growth hormone (GH) secretion from somatotropes in the pituitary of the goldfish Carassius auratus was investigated. Immunocytochemistry with two antibodies against mammalian NO synthase (NOS) revealed the presence of a NOS-like enzyme in primary cultures of dispersed goldfish pituitary cells, including morphologically identified somatotropes. NO donors S-nitroso-N-acetylpenicillamine and sodium nitroprusside (SNP), as well as a cyclic guanosine monophosphate analogue (dibutyryl guanosine 3':5'-cyclic monophosphate), all significantly increased GH secretion from dispersed goldfish pituitary cells in static culture. Somatostatin abolished the response to SNP, and NOS inhibitors aminoguanadine hemisulfate (AGH) and N-(3 aminomethyl)benzylacetamidine, dihydrochloride (1400W) decreased the GH release response to known neuroendocrine factors stimulatory to GH release (gonadotropin releasing hormone and a dopamine D1 agonist). AGH and 1400W did not alter basal GH secretion. These data suggest that NO plays a role in mediating the GH response to endogenous neuroendocrine factors in goldfish. PMID- 10843798 TI - Remodeling of turkey adrenal steroidogenic tissue induced by dietary protein restriction: the potential role of cell death. AB - The present study focused on the cellular remodeling of steroidogenic tissue in the domestic turkey (Meleagris gallopavo) adrenal gland in response to dietary protein restriction stress. Immature male turkeys (1 week old) were fed isocaloric synthetic diets containing either 28% (control) or 8% (restriction) soy protein for 4 weeks. Adrenal glands were processed for the isolation of density- separable, visibly distinct adrenal steroidogenic cell subpopulations: three low-density subpopulations [LDAC-1 (rho = 1. 0350-1.0490 g/ml), LDAC-2 (rho = 1.0490-1.0570 g/ml), and LDAC-3 (rho = 1.0570-1.0585 g/ml)] and one high density subpopulation [HDAC (rho = 1.0590-1.0720 g/ml)]. Dietary protein restriction increased the proportion of LDAC-3 and HDAC by 98 and 350%, respectively, and decreased LDAC-2 by 46%. LDAC-1 also showed signs of proportional decrement. To determine the role of cell death in this process, the potential for apoptosis was assessed in adrenal tissue and isolated adrenal steroidogenic cells using short-term culture followed by analysis of oligonucleosome formation. Basal, culture-triggered oligonucleosome formation of tissue and cells derived from protein-restricted birds was 80% greater than that of tissue and cells derived from control birds. This differential in apoptotic potential persisted with a variety of treatments, in vitro. Apoptotic potential was suppressed by human adrenocorticotropin and enhanced by angiotensin II (Ang II). The proapoptotic effect of Ang II (100 nM) with adrenal fragments was inhibited by the Ang II receptor antagonist [Sar(1), Ile(8)]ang II (10 microM) to below basal values (by about 60%), but the inhibition was surmountable by high concentrations (10 and 100 microM) of Ang II. The antagonist also attenuated basal, culture-triggered DNA fragmentation of tissue and cells, suggesting that at least part of the basal DNA fragmentation was due to intrinsically generated Ang II. Differences in apoptotic potential were also apparent with cell subpopulations. Compared to control subpopulations, protein restriction enhanced basal oligonucleosome formation in LDAC-1 and -2 by 38 and 122%, respectively, and reduced it in LDAC-3 and HDAC by 53 and 70%, respectively. These data suggest a role for apoptotic cell death in the remodeling of turkey adrenal steroidogenic tissue induced by dietary protein restriction. In addition, other data suggest that Ang II is an important regulator of adrenal steroidogenic cell turnover in the avian adrenal gland. PMID- 10843799 TI - Identification of differentially expressed genes in cardiac hypertrophy by analysis of expressed sequence tags. AB - Cardiac hypertrophy is an adaptive response to chronic hemodynamic overload. We employed a whole-genome approach using expressed sequence tags (ESTs) to characterize gene transcription and identify new genes overexpressed in cardiac hypertrophy. Analysis of general transcription patterns revealed a proportional increase in transcripts related to cell/organism defense and a decrease in transcripts related to cell structure and motility in hypertrophic hearts compared to normal hearts. Detailed comparison of individual gene expression identified 64 genes potentially overexpressed in hypertrophy, of 232 candidate genes derived from a set of 77,692 cardiac ESTs, including 47,856 ESTs generated in our laboratory. Of these, 29 were good candidates (P < 0.0002) and 35 were weaker candidates (P < 0.005). RT-PCR of a number of these candidate genes demonstrated correspondence of EST-based predictions of gene expression with in vitro levels. Consistent with an organ under various stresses, up to one-half of the good candidates predicted to exhibit differential expression were genes potentially involved in stress response. Analyses of general transcription patterns and of single-gene expression levels were also suggestive of increased protein synthesis in the hypertrophic myocardium. Overall, these results depict a scenario compatible with current understanding of cardiac hypertrophy. However, the identification of several genes not previously known to exhibit increased expression in cardiac hypertrophy (e.g., prostaglandin D synthases; CD59 antigen) also suggests a number of new avenues for further investigation. These data demonstrate the utility of genome-based resources for investigating questions of cardiovascular biology and medicine. PMID- 10843800 TI - Chromosome translocations in breast cancer with breakpoints at 8p12. AB - Unbalanced chromosome translocations with breakpoints around 8p12, resulting in loss of distal 8p, are common in carcinomas. We have mapped the 8p12 breakpoints in three breast cancer cell lines, T-47D, MDA-MB-361, and ZR-75-1, using YACs and PACs between D8S540 and D8S255 by fluorescence in situ hybridization. All three lines had a breakpoint close to D8S505, proximal to HGL. Each breakpoint was distinct, but all were within 0.5 to 1.5 Mb of each other. The T-47D cell line had a straightforward translocation, but in MDA-MB-361 and ZR-75-1 the translocations were accompanied by local rearrangements of surprising complexity. Small regions of 8p from close to the breakpoint were duplicated or amplified as inserts in the attached chromosome fragment. ZR-75-1 also had retained a separate fragment of about 1 Mb, from the region 1 to 3 Mb telomeric to the common breakpoint, that included HGL. This line also had an interstitial deletion several megabases more centromeric. The data suggest that breakpoints on 8p12 are clustered in a small region and show that translocations breaking there may be accompanied by additional rearrangements. PMID- 10843801 TI - Transcription mapping of the 5q- syndrome critical region: cloning of two novel genes and sequencing, expression, and mapping of a further six novel cDNAs. AB - The 5q- syndrome is a myelodysplastic syndrome with the 5q deletion ?del(5q) as the sole karyotypic abnormality. We are using the expressed sequence tag (EST) resource as our primary approach to identifying novel candidate genes for the 5q- syndrome. Seventeen ESTs were identified from the Human Gene Map at the National Center for Biotechnology Information that had no significant homology to any known genes and were assigned between DNA markers D5S413 and D5S487, flanking the critical region of the 5q- syndrome at 5q31-q32. Eleven of the 17 cDNAs from which the ESTs were derived (65%) were shown to map to the critical region of the 5q- syndrome by gene dosage analysis and were then sublocalized by PCR screening to a YAC contig encompassing the critical region. Eight of the 11 cDNA clones, upon full sequencing, had no significant homology to any known genes. Each of the 8 cDNA clones was shown to be expressed in human bone marrow. The complete coding sequence was obtained for 2 of the novel genes, termed C5orf3 and C5orf4. The 2.6 kb transcript of C5orf3 encodes a putative 505-amino-acid protein and contains an ATP/GTP-binding site motif A (P loop), suggesting that this novel gene encodes an ATP- or a GTP-binding protein. The novel gene C5orf4 has a transcript of 3.1 kb, encoding a putative 144-amino-acid protein. We describe the cloning of 2 novel human genes and the sequencing, expression patterns, and mapping to the critical region of the 5q- syndrome of a further 6 novel cDNA clones. Genomic localization and expression patterns would suggest that the 8 novel cDNAs described in this report represent potential candidate genes for the 5q- syndrome. PMID- 10843802 TI - Construction of a 350-kb sequence-ready 11q13 cosmid contig encompassing the markers D11S4933 and D11S546: mapping of 11 genes and 3 tumor-associated translocation breakpoints. AB - Previously, we located three novel human tumor-associated translocation breakpoints in the chromosome 11q13 region between the markers D11S4933 and D11S546. To facilitate the molecular analysis of these breakpoints, we have constructed a continuous sequence-ready cosmid and PAC contig of approximately 350 kb, including the markers D11S4933 and D11S546. In addition, a detailed transcript map was generated. This resulted in the precise positioning of 11 genes and ESTs within the contig, including 4 genes already known to map in the 11q13 region. Three other genes that we positioned within the contig showed homologies to unmapped genes from human and/or other species. Three ESTs were novel. Partial cosmid sequencing resulted in the establishment of the direction of transcription of several of the reported genes. This contig will be instrumental for the detailed characterization of the tumor-associated chromosomal breakpoints and the identification of other 11q13-associated disease genes. PMID- 10843803 TI - Genomic structure, expression, and chromosomal localization of the human glycine N-methyltransferase gene. AB - The glycine N-methyltransferase (GNMT) gene encodes a protein that not only acts as an enzyme to regulate the ratio of S-adenosylmethionine to S adenosylhomocysteine, but also participates in the detoxification pathway in liver cells. Previously, we reported that the expression level of GNMT was diminished in human hepatocellular carcinoma. In this study, the human GNMT gene was cloned and characterized. It contains six exons and spans about 10 kb. Instead of a TATA box, it has a transcriptional initiator located 801 bp upstream from the translation start codon. The gene was localized to chromosome 6p12 using fluorescence in situ hybridization. Northern blot analysis of 16 tissues from different human organs showed that GNMT was expressed only in liver, pancreas, and prostate. PMID- 10843804 TI - Identification and characterization of YME1L1, a novel paraplegin-related gene. AB - A gene responsible for an autosomal recessive form of hereditary spastic paraplegia (SPG7) was recently identified. This gene encodes paraplegin, a mitochondrial protein highly homologous to the yeast mitochondrial AAA proteases Afg3p, Rca1p, and Yme1p, which have both proteolytic and chaperone-like activities at the inner mitochondrial membrane. By screening the expressed sequence tag database, we identified and characterized a novel human gene, YME1L1 (YME1L1-like1, HGMW-approved symbol). This gene encodes a predicted protein of 716 amino acids highly similar to all mitochondrial AAA proteases and in particular to yeast Yme1p. Expression and immunofluorescence studies revealed that YME1L1 and paraplegin share a similar expression pattern and the same subcellular localization in the mitochondrial compartment. YME1L1 may represent a candidate gene for other forms of hereditary spastic paraplegia and possibly for other neurodegenerative disorders. PMID- 10843805 TI - A high-resolution radiation hybrid map of the proximal portion of mouse chromosome 5. AB - Radiation hybrid (RH) mapping of the mouse genome provides a useful tool in the integration of existing genetic and physical maps, as well as in the ongoing effort to generate a dense map of expressed sequence tags. To facilitate functional analysis of mouse Chromosome 5, we have constructed a high-resolution RH map spanning 75 cM of the chromosome. During the course of these studies, we have developed RHBase, an RH data management program that provides data storage and an interface to several RH mapping programs and databases. We have typed 95 markers on the T31 RH panel and generated an integrated map, pooling data from several sources. The integrated RH map ranges from the most proximal marker, D5Mit331 (Chromosome Committee offset, 3 cM), to D5Mit326, 74.5 cM distal on our genetic map (Chromosome Committee offset, 80 cM), and consists of 138 markers, including 89 simple sequence length polymorphic markers, 11 sequence-tagged sites generated from BAC end sequence, and 38 gene loci, and represents average coverage of approximately one locus per 0.5 cM with some regions more densely mapped. In addition to the RH mapping of markers and genes previously localized on mouse Chromosome 5, this RH map places the alpha-4 GABA(A) receptor subunit gene (Gabra4) in the central portion of the chromosome, in the vicinity of the cluster of three other GABA(A) receptor subunit genes (Gabrg1-Gabra2-Gabrb1). Our mapping effort has also defined a new cluster of four genes in the semaphorin gene family (Sema3a, Sema3c, Sema3d, and Sema3e) and the Wolfram syndrome gene (Wfs1) in this region of the chromosome. PMID- 10843806 TI - Molecular characterization of KLHL3, a human homologue of the Drosophila kelch gene. AB - The Drosophila kelch protein is a structural component of ring canals and is required for oocyte maturation. Here, we report the cloning and genomic structure of a new human homologue of kelch, KLHL3. At the amino acid level, KLHL3 shares 77% similarity with Drosophila kelch and 89% similarity with Mayven (KLHL2), another human kelch homolog. The approximately 6.5-kb mRNA has a single open reading frame encoding a protein of 587 amino acids with a predicted molecular mass of 650 kDa. Like kelch and KLHL2, the KLHL3 protein contains a poxvirus and zinc finger domain at the N-terminus and six tandem repeats (kelch repeats) at the C-terminus. At least three isoforms, which differ in the length of the N terminus, are produced and may be the result of alternative promoter usage. We also identified alternative polyadenylation sites and alternative splicing; thus, as many as 12 mRNA variants and six putative protein isoforms could be produced. The KLHL3 gene is mapped to human chromosome 5, band q31, contains 17 exons, and spans approximately 120 kb of genomic DNA. KLHL3 maps within the smallest commonly deleted segment in myeloid leukemias characterized by a deletion of 5q; however, we detected no inactivating mutations of KLHL3 in malignant myeloid disorders with loss of 5q. PMID- 10843807 TI - The ancient source of a distinct gene family encoding proteins featuring RING and C(3)H zinc-finger motifs with abundant expression in developing brain and nervous system. AB - Intronless genes can arise by germline retrotransposition of a cDNA originating as mRNA from an intron-containing source gene. Previously, we described several members of a family of intronless mammalian genes encoding a novel class of zinc finger proteins, including one that shows imprinted expression and one that escapes X-inactivation. We report here the identification and characterization of the Makorin ring finger protein 1 gene (MKRN1), a highly transcribed, intron containing source for this family of genes. Phylogenetic analyses clearly indicate that the MKRN1 gene is the ancestral founder of this gene family. We have identified MKRN1 orthologs from human, mouse, wallaby, chicken, fruitfly, and nematode, underscoring the age and conservation of this gene. The MKRN gene family encodes putative ribonucleoproteins with a distinctive array of zinc finger motifs, including two to four C(3)H zinc-fingers, an unusual Cys/His arrangement that may represent a novel zinc-finger structure, and a highly conserved RING zinc-finger. To date, we have identified nine MKRN family loci distributed throughout the human genome. The human and mouse MKRN1 loci map to a conserved syntenic group near the T-cell receptor beta cluster (TCRB) in chromosome 7q34-q35 and chromosome 6A, respectively. MKRN1 is widely transcribed in mammals, with high levels in murine embryonic nervous system and adult testis. The ancient origin of MKRN1, high degree of conservation, and expression pattern suggest important developmental and functional roles for this gene and its expressed family members. PMID- 10843808 TI - Significant evidence for linkage disequilibrium over a 5-cM region among Afrikaners. AB - We explore the extent of deviations from Hardy-Weinberg equilibrium (HWE) at a marker locus and linkage disequilibrium (LD) between pairs of marker loci in the Afrikaner population of South Africa. DNA samples were used for genotyping of 23 loci on six chromosomes. The samples were collected from 91 healthy unrelated Afrikaner adults. Exact tests were used to determine evidence for deviations from HWE at a single marker locus or LD between pairs of marker loci. At the 0.05 level of significance, evidence was found for deviation from HWE at only one of the 23 loci. At the same level of significance, LD was found among 8 of the 34 intrachromosomal pairs of loci. On chromosome 21, there was evidence for LD (P = 0.02) between a pair of loci with a genetic distance of 5.51 cM. On chromosome 2, there was evidence for LD between a pair of loci with a genetic distance of 5.28 cM (P = 0.002) and a pair of loci with a genetic distance of 3.68 cM (P = 0.0004). Detailed analysis of LD for one locus pair indicated that only a few of all alleles participated in the LD and that strong LD was most often positive. Our findings indicate that Afrikaans-speaking Afrikaners represent one of those special populations deemed particularly suitable for disequilibrium mapping. PMID- 10843809 TI - A common set of at least 11 functional genes is lost in the majority of NF1 patients with gross deletions. AB - Large deletions of the NF1 locus occur in 5 to 10% of patients with neurofibromatosis and are commonly associated with specific additional abnormalities characterized by mental retardation, dysmorphic features, and intellectual impairment. To characterize the extent of codeleted genes we constructed a long-range physical BAC/PAC map around the NF1 locus between D17S117 and D17S57 and determined the deletion boundaries in seven unrelated patients. Surprisingly, the proximal and distal breakpoints in five of seven patients fall at almost identical positions, resulting in the loss of at least 11 functional genes. Five of six patients investigated showed a de novo deletion on the maternally derived chromosome. Since D17S117 and D17S57 were previously reported as the outer limits for the great majority of NF1 deletions, we suggest that most NF1 patients with deletion of the entire NF1 gene are hemizygous for the same set of at least 10 additional genes, including SHGC-37343, SHGC-2390, SHGC-34232, OMG, EVI2B, EVI2A, WI-9521, WI-6742, SHGC-34334, and KIAA0160, and thus present with a relatively uniform clinical phenotype. PMID- 10843810 TI - HERV-K-T47D-Related long terminal repeats mediate polyadenylation of cellular transcripts. AB - The human genome harbors thousands of long terminal repeats (LTRs) that are derived from endogenous retroviruses and contain elements able to regulate the expression of neighboring cellular genes. We have investigated the ability of human endogenous retroviral (HERV)-K LTRs to provide transcriptional processing signals for nonviral sequences. Four chimeric cDNA clones isolated from a cDNA library derived from the human cell line T47D were found to be polyadenylated within an HERV-K-T47D-related LTR. Two transcripts containing an as yet unknown cellular sequence were probably derived from the same genomic locus but their 3' ends were processed at different positions of the LTR. Structural analysis of the polyadenylation site suggests RNA stem-loop structures similar to the HTLV-1 Rex responsive element that bring the two remote AAUAAA and GU-rich elements into the spatial juxtaposition necessary for correct 3' end processing. The cellular part of the third chimeric clone shows significant homology to an exon of the human tyrosine phosphatase 1 gene, although oriented in the antisense direction compared to the adjacent LTR. Furthermore, we found that the 3' untranslated region of the human transmembrane tyrosine kinase gene FLT4 is probably derived from a partial HERV-K-T47D LTR sequence. Taken together, our data suggest that LTRs of the HERV-K-T47D family display biological function by mediating polyadenylation of cellular sequences. PMID- 10843811 TI - Large-insert clone/STS contigs in Xq11-q12, spanning deletions in patients with androgen insensitivity and mental retardation. AB - An integrated large-insert clone map of the region Xq11-q12 is presented. A physical map containing markers within a few hundred kilobases of the centromeric locus DXZ1 to DXS1125 spans nearly 5 Mb in two contigs separated by a gap estimated to be approximately 100-250 kb. The contigs combine 75 yeast artificial chromosome clones, 12 bacterial artificial chromosome clones, and 17 P1-derived artificial chromosome clones with 81 STS or EST markers. Overall marker density across this region is approximately 1 STS/60 kb. Mapped within the contigs are 12 ESTs as well as 5 known genes, moesin (MSN), hephaestin (HEPH), androgen receptor (AR), oligophrenin-1 (OPHN1), and Eph ligand-2 (EPLG2). Orientation of the contigs on the X chromosome, as well as marker order within the contigs, was unambiguously determined by reference to a number of X chromosome breakpoints. In addition, the distal contig spans deletions from chromosomes of three patients exhibiting either complete androgen insensitivity (CAI) or a contiguous gene syndrome that includes CAI, impaired vision, and mental retardation. PMID- 10843812 TI - Physical map of the region surrounding the OTOFERLIN locus on chromosome 2p22 p23. AB - The autosomal recessive form of nonsyndromic deafness DFNB9 has been mapped to a 2-cM region on chromosome 2p22-p23, and the responsible gene, OTOF, has been recently identified by positional cloning combined with a candidate gene approach. In the course of this gene cloning, we established a contig of yeast artificial chromosomes, bacterial artificial chromosomes, and P1 phage artificial chromosomes delimited by polymorphic markers D2S2170 and D2S170, i.e. , extending over approximately 3500 kb. Sixty expressed sequence tags or genes and 14 sequence-tagged sites, 11 of which are polymorphic, were mapped to this contig and assigned to 21 chromosomal intervals. PMID- 10843813 TI - Cloning of the cDNA and localization of the gene encoding human NRBP, a ubiquitously expressed, multidomain putative adapter protein. AB - Adapter proteins modulate multiple signaling pathways by regulating the aggregation of other factors into signaling complexes. Here we have identified a novel human cDNA encoding NRBP, a multidomain putative adapter protein containing (i) two putative nuclear receptor binding motifs (LXXLL), (ii) a putative binding domain for Src homology-2 (SH2) domain containing proteins, (iii) a kinase-like domain, (iv) a bipartite nuclear localization signal, and (v) three sequences rich in glutamic acid, serine, proline, and threonine (PEST) residues. The NRBP mRNA transcript, of approximately 2.4 kb, was ubiquitously expressed in a wide range of normal human tissues and 15 human tumor cell lines. The NRBP cDNA is predicted to encode a polypeptide of 535 amino acids with a molecular mass of 59.8 kDa. Translation of NRBP mRNA in vitro reveals three translation products of 60, 51, and 43 kDa, suggesting that translation of NRBP may initiate at multiple sites. The NRBP gene was localized to human chromosome 2p23, near the location of the NCOA1 gene encoding the nuclear receptor coactivator, steroid receptor coactivator-1 (SRC-1). The features of NRBP predict a function as an adapter protein potentially linking signaling pathways involving nuclear receptors and SH2 domain containing proteins. PMID- 10843814 TI - Confirmation of the mapping of the Camurati-Englemann locus to 19q13. 2 and refinement to a 3.2-cM region. AB - Camurati-Englemann syndrome (DPD1) is an autosomal dominant condition associated with progressive cortical sclerosis of the diaphyses of all the long bones. Clinical features include abnormal gait, muscle weakness and wasting, and generalized fatigue. The DPD1 gene was recently mapped to a 15.1-cM region on chromosome 19q13.2. We have narrowed the region containing the DPD1 gene to a 3.2 cM region flanked by short tandem repeat markers, D19S881 and D19S718. TGFB1, a candidate gene mapped within this region, was excluded. PMID- 10843815 TI - Is a mechanical or a metabolic approach superior in the treatment of coronary disease? Results of the atorvastatin versus revascularization (AVERT) trial. PMID- 10843816 TI - Aortic valve replacement: an update at the turn of the millennium. PMID- 10843817 TI - Digoxin-like immunoreactive substances in hypertrophic cardiomyopathy. PMID- 10843818 TI - Relationship of serum C3 complement with insulin resistance and coronary heart disease-cause, consequence or common antecedent? PMID- 10843819 TI - Myocardial viability: the search for a perfect method is not over yet. PMID- 10843820 TI - Evaluating risk in unstable angina: role of pharmacological stress echocardiography. PMID- 10843821 TI - Coronary artery disease, depression and social support only the beginning. PMID- 10843822 TI - Chlamydia pneumonia (and other infective agents) in atherosclerosis and acute coronary syndromes. How good is the evidence? PMID- 10843823 TI - The natural history of prevalent ischaemic heart disease in middle-aged men. AB - OBJECTIVE: To describe the long-term outcome of different forms of symptomatic and asymptomatic ischaemic heart disease in middle-aged men. METHODS: 7735 men aged 40-59, randomly selected from 24 general practices in Britain were classified into one of seven ischaemic heart disease groups according to a questionnaire and electrocardiogram (ECG): I=diagnosed myocardial infarction; II=unrecognized myocardial infarction; III= diagnosed angina; IV=angina symptoms; V=possible myocardial infarction symptoms; VI=ECG ischaemia or possible myocardial infarction; VII=no evidence of ischaemic heart disease. The association of disease group with a range of fatal and non-fatal outcomes during 15 years of follow-up was assessed. RESULTS: At baseline 25% of men had evidence of ischaemic heart disease (groups I-VI). Risks of major ischaemic heart disease events, total and cardiovascular mortality, stroke, and major cardiovascular events tended to increase strongly from group VII to I. Diagnosed myocardial infarction was associated with a much poorer prognosis than all other groups (including unrecognized infarction) for all cardiovascular outcomes other than stroke. The relative risk associated with ischaemic heart disease at baseline declined dramatically over time. However, men with myocardial infarction who survived event-free for 10 years continued to experience a high excess risk in the subsequent 5 years, in contrast to event-free survivors of angina and other ischaemic heart disease. Adjusted to an average age of 50, the percentage of men surviving for 15 years free of a new major cardiovascular event was 44 for diagnosed myocardial infarction, 52 for unrecognized myocardial infarction, 66 for diagnosed angina, 68 for angina symptoms, 73 for possible myocardial infarction symptoms, 73 for ECG ischaemia, and 79 for no ischaemic heart disease. Comparison of outcome between prevalent and incident myocardial infarction illustrated the improved prognosis of men surviving the initial years after their event. CONCLUSIONS: Differing manifestations of prevalent ischaemic heart disease are associated with widely differing outcome, and the majority of middle-aged men in the community who have evidence of ischaemic heart disease short of myocardial infarction survive for 15 years without heart attack or stroke. The excess risk associated with myocardial infarction appears more persistent than that associated with angina and other ischaemic heart disease, remaining high even after 10 years of event-free survival. PMID- 10843824 TI - Feasibility and prognostic value of dobutamine-atropine stress echocardiography early in unstable angina. AB - BACKGROUND AND AIM: Because unstable angina has always been considered a contraindication for dobutamine-atropine stress echocardiography (DSE), the role of dobutamine-atropine stress echocardiography in unstable angina is unknown. Our aim was to assess the safety and prognostic value of dobutamine-atropine stress echocardiography in unstable angina. METHODS: One hundred and thirty-two patients were studied (mean age 64+/-12 years, 29 women). Dobutamine-atropine stress echocardiography was performed on the third day after hospital admission. End points were unstable angina, myocardial infarction or cardiac death at 1 year follow-up. RESULTS: No major complications occurred during dobutamine-atropine stress echocardiography. Ninety-six (78%) patients were on beta-blocker therapy during the test; mean maximum heart rate achieved was 106+/-23 beats x min(-1). Nine of the 21 patients (43%) with a positive dobutamine-atropine stress echocardiography presented cardiac events during follow-up: two patients died, one had a myocardial infarction and six had recurrent class III-IV angina. Among 80 patients with negative dobutamine-atropine stress echocardiography, one (1%) had myocardial infarction and six patients (7.5%) had recurrent angina. Event free survival after 1 year for patients with a negative dobutamine-atropine stress echocardiography for ischaemia was 91% compared to 57% for those with a positive dobutamine-atropine stress echocardiography (P<0. 0001). Left ventricular dysfunction (P=0.01), prior myocardial infarction (P=0.03) and a positive dobutamine-atropine stress echocardiography (P=0.004) were independent predictors of cardiac events during follow-up. CONCLUSIONS: Dobutamine-atropine stress echocardiography is safe in unstable angina if it is performed when patients remain asymptomatic for at least 48 h. A negative dobutamine-atropine stress echocardiogram for ischaemia predicts a good prognosis in medically treated patients with unstable angina and may allow their early discharge from hospital. Good prognostic information was obtained despite the use of beta blockers and low heart rates during dobutamine-atropine stress echocardiography. PMID- 10843825 TI - Depressive symptoms and lack of social integration in relation to prognosis of CHD in middle-aged women. The Stockholm Female Coronary Risk Study. AB - AIMS: Several studies have reported that women with coronary heart disease have a poorer prognosis than men. Psychosocial factors, including social isolation and depressive symptoms have been suggested as a possible cause. However, little is known about these factors and their independent predictive value in women. Therefore, we investigated the prognostic impact of depression, lack of social integration and their interaction in the Stockholm Female Coronary Risk Study. METHODS AND RESULTS: Two hundred and ninety-two women patients aged 30 to 65 years and admitted for an acute coronary event between 1991 and 1994, were followed for 5 years from baseline assessments, which were performed between 3 and 6 months after admission. Lack of social integration and depressive symptoms, assessed at baseline by standardized questionnaires, were associated with recurrent events, including cardiovascular mortality, acute myocardial infarction and revascularization procedures (percutaneous transluminal coronary angioplasty and coronary artery bypass grafting). Adjusting for age, diagnosis at index event, symptoms of heart failure, diabetes mellitus, high density lipoprotein (HDL) cholesterol, history of hypertension, systolic blood pressure, smoking, sedentary lifestyle, body mass index, and severity of angina pectoris symptoms, the hazard ratio associated with low (lowest quartile) as compared to high social integration (upper quartile) was 2.3 (95% CI 1.2-4.5) and the hazard ratio associated with two or more (upper three quartiles) as compared to one or no depressive symptoms was 1.9 (95% CI 1.02-3.6). CONCLUSIONS: The presence of two or more depressive symptoms and lack of social integration independently predicted recurrent cardiac events in women with coronary heart disease. Women who were free of both these risk factors, had the best prognosis. PMID- 10843826 TI - Relationship of serum C3 to fasting insulin, risk factors and previous ischaemic events in middle-aged men. AB - AIMS: Serum C3 is a powerful indicator of the risk of myocardial infarction, which correlates with body mass index, serum lipids and blood pressure. This study was performed to ascertain whether such correlations may be explained by an association of C3 with fasting insulin, and to assess comparatively the relationships of C3 and traditional risk factors to previous myocardial infarction. METHODS AND RESULTS: The fasting levels of C3, insulin, and the main risk factors were evaluated in 1090 unselected men aged 55-64 years, including 129 cases of previous ischaemic events (51 myocardial infarctions). In multivariate analysis C3 was associated with insulin (r=0.27, P<0.0001), cholesterol (r=0.18, P<0.0001), body mass index (r=0.13, P<0.0001), glucose (r=0.12, P=0.0001), systolic blood pressure (r=0.10, P<0.001), triglycerides (r=0.09, P<0.01) and HDL-cholesterol (r=-0.06, P<0.05). These variables explained 31% of the total C3 variance. Alcohol consumption and physical activity correlated inversely with C3, while no correlation was found with smoking and family history of myocardial infarction. C3 was associated with previous myocardial infarction and stroke, but not with angina pectoris and peripheral arterial disease. In logistic regression the variables associated with previous myocardial infarction were C3 (P=0.011), family history of myocardial infarction (P=0.018), ex-smoker status (P=0.020), age (P=0.025), glucose (P=0.028) and HDL cholesterol (P=0.051, inverse relationship). CONCLUSIONS: The association of C3 with myocardial infarction persists retrospectively, and is more significant than any other association of traditional risk factors with previous myocardial infarction. Of the many variables associated with C3, fasting insulin is its main covariate, which suggests that C3 is a marker of a pro-atherogenic metabolic imbalance partly coinciding with insulin resistance. PMID- 10843827 TI - Doppler tissue velocity sampling improves diagnostic accuracy during dobutamine stress echocardiography for the assessment of viable myocardium in patients with severe left ventricular dysfunction. AB - BACKGROUND: Both nuclear imaging with F18-fluorodeoxyglucose and dobutamine stress echocardiography have been used to identify viable myocardium, although dobutamine-stress echocardiography has been demonstrated to be the less sensitive of the two. AIM: To compare the accuracy of pulsed-wave Doppler tissue sampling with dobutamine-stress echocardiography for the detection of viable myocardium, using F18-fluorodeoxyglucose imaging as a reference. Methods Forty patients with chronic coronary artery disease and left ventricular dysfunction (mean ejection fraction 33+/-11%), underwent F18-fluorodeoxyglucose imaging, dobutamine-stress echocardiography and pulsed-wave Doppler tissue sampling. Evaluation was performed using a six-segment model. RESULTS: Visual assessment by resting echo was feasible in 230 out of 240 segments (96%); 177 (77%) segments showed severe dyssynergy at rest. F18-fluorodeoxyglucose imaging showed viability in 95 (54%) segments while 82 (46%) were non-viable. Ejection phase velocity at rest was not significantly different; ejection velocities during low-dose and peak-dose dobutamine, however, were significantly higher in viable myocardium (8.6+/-2.9 vs 6.0+/-1.8 and 9.3+/-3.1 vs 6.2+/-2.1 cm x s(-1)). Using receiver operating characteristic curves the optimal cut-off value for viability assessment was an increase in the ejection phase velocity low-dose of 1+/-0.5 cm x s(-1), while 0+/ 0.5 cm x s(-1)predicted non-viability. The sensitivity and specificity (95%CI) of pulsed-wave Doppler tissue sampling and dobutamine-stress echocardiography for the prediction of viability was respectively 87% (82-92) vs 75% (67-81) (P<0.05) and 52% (44-59) vs 51% (45-59) (P=ns). CONCLUSIONS: The sensitivity of pulsed wave Doppler tissue sampling is superior to dobutamine-stress echocardiography for the assessment of myocardial viability. PMID- 10843829 TI - ESC News and Appointments. PMID- 10843828 TI - Long-term follow-up of morbidity and mortality after aortic valve replacement with a mechanical valve prosthesis. AB - AIMS: The aim of this study was to determine the incidence of valve-related complications in patients with a mechanical aortic valve prosthesis and to identify risk factors for an adverse outcome. METHODS AND RESULTS: In the 424 patients, event-free survival rates 5 and 10 years after aortic valve replacement were 62% and 37%, respectively. The linearized incidence of thromboembolic events was 4.4% per patient-year, and of anticoagulant-related haemorrhage 8.5% per patient-year. Advanced NYHA functional class, atrial fibrillation, pure aortic regurgitation and thromboembolism prior to surgery decreased event-free survival. A history of pre-operative thromboembolism increased the risk for a first embolic event after aortic valve replacement (relative hazard [RH] 3.2), but was even more strongly associated with the risk for repeated events (> or =2 events, RH 5.4). After each thromboembolic episode that occurred, the risk for a subsequent one was increased. The risk for at least one, and up to three or more haemorrhages was increased in patients with a pre-operative history of bleeding (RH 3.3-5.1) and of atrial fibrillation (RH 1.8-3.9). The risk for a subsequent event was increased by a history of repeated haemorrhages, a short interval since previous bleeding, and high age. CONCLUSIONS: There were few factors strongly related to valve related morbidity. However, previous bleedings and previous thromboembolism were powerful risk factors for repeated events. PMID- 10843830 TI - Sedimentation of Bacillus sphaericus in tap water and sewage water. AB - Formulations of Bacillus sphaericus products were applied in 1-m columns of clean water and sewage water in tubes. Samples taken through valves attached to the walls of the tubes showed that spores settled as a function of application method, droplet size, particle size, and water characteristics. In a mixture of deionized and tap water, the slowest sedimentation rate was obtained with a primary powder sonicated in water with 0.1% detergent. This product and a commercial fluid product settled faster in sewage water than in clean water and after 4 weeks, most spores were found in the sediment. In clean water, most spores remained suspended during the 4 weeks but the density was highest at the bottom. Calculation of total spore numbers in the columns after 4 weeks indicated that about 90% of spores disappeared in clean water, and 95-97% disappeared in sewage water. Bioassays showed that toxicity declined in parallel with the sedimentation, but at a higher rate, indicating a loss of toxicity from the residual spores. PMID- 10843831 TI - Susceptibility of Vespula vulgaris (Hymenoptera: vespidae) to generalist entomopathogenic fungi and their potential for wasp control. AB - The pathogenicity of Vespula vulgaris wasp workers and larvae to a range of fungi was determined. All fungi were isolated in New Zealand and included isolates from Vespula, known generalist insect pathogens, and isolates generally nonpathogenic to insects. Workers and larvae were highly susceptible to pathogenic isolates at high spore concentrations (>1.75 x 10(5) cfu/individual). Eight isolates, two of Metarhizium anisopliae, five of Beauveria bassiana, and one of Aspergillus flavus were pathogenic while a single isolate of M. flavouiride var. novazealandicum, Cladosporium sp., and Paecilomyces sp. were not. The transfer of spores between workers, and between workers and larvae, was also investigated using several different application methods. Transfer of spores occurred between treated and untreated individuals, and for some of the application methods sufficient spores were transferred to cause mortality of the nontreated individuals. These findings are related to the potential of fungi for the control of wasps. PMID- 10843832 TI - The interactions between soybean trypsin inhibitor and delta-endotoxin of Bacillus thuringiensis in Helicoverpa armigera larva. AB - No significant difference in larval mortality was observed when a sublethal dose of Bacillus thuringiensis (Bt) var. kurstaki HD-1 crystal was supplemented with soybean trypsin inhibitor (STI) in the artificial diet fed to Helicoverpa armigera in the laboratory, but supplementing a nonlethal dose of crystal with STI in the diet led to a pronounced reduction of larval growth. This concentration of crystal and two lower concentrations of STI alone had no significant effects on larval growth. The results of substrate-gel electrophoresis demonstrated that the proteases in the H. armigera midgut fluid responsible for the degradation of protoxin consisted of at least four proteases with molecular weights of 71, 49, 36, and 30 kDa. All four proteases could utilize casein also as the substrate. When larvae were fed with STI or Bt + STI, the proteolytic activities of the 49-kDa enzyme disappeared, and the activities of the other three enzymes were reduced. Enzyme assays also indicated that feeding larvae with diets containing Bt, STI, or Bt + STI significantly decreased the specific activities of larval general proteases and the trypsin-like enzyme. The protein concentration of midgut fluid was elevated, especially in the larvae fed on the diets containing STI and Bt + STI. Both in vitro and in vivo studies showed that the degradation of protoxin and toxin could be inhibited by soybean trypsin inhibitors, but when the incubation time was prolonged, the protoxin could be degraded completely, while the degradation of toxin was inhibited further. This suggested that the retention time of toxins in the larval midgut was extended and synergism between insecticidal crystal protein and soybean trypsin inhibitor occurred, which showed as the inhibition of H. armigera larval growth. PMID- 10843833 TI - Wolbachia density and host fitness components in Muscidifurax uniraptor (Hymenoptera: pteromalidae). AB - Intracellular bacteria of the genus Wolbachia are found in a variety of arthropod hosts, where they cause various reproductive disorders. Attempts to study the fitness advantages and disadvantages of carrying these symbionts have yielded contradicting results. Using various doses of the antibiotic rifampicin, we were able to manipulate the density of Wolbachia in the uniparental parasitoid Muscidifurax uniraptor (Hymenoptera: Pteromalidae). The effect of different titers of the symbiont on the fecundity, reproductive rate, longevity, survival rate, and sex ratio of the host was measured. The data gathered show that following antibiotic treatments, the percentage of males rises at low doses of rifampicin and then drops again. The total sex ratio of offspring produced by treated mothers was positively correlated with the numbers of Wolbachia found in eggs laid by these females. No significant effects were detected with regard to the other studied fitness components. It is concluded that in M. uniraptor, Wolbachia are not posing any burden on the life history trait studied. PMID- 10843834 TI - Characterization of a granulovirus from the cassava hornworm (Erinnyis ello: Sphingidae). AB - A Colombian isolate of Erinnyis ello granulovirus (EeGV) was characterized by electron microscopy, restriction endonuclease digestion, and SDS-PAGE. Electron microscopy showed the occlusion bodies to have a morphology typical of granuloviruses. The restriction patterns of DNA from EeGV and the granuloviruses of Trichoplusia ni (TnGV) and Pieris rapae (PrGV) show little or no similarity, indicating little relatedness among these viruses. EeGV was estimated to possess a relatively small genome of 90.5 +/- 0.5 kbp. SDS-PAGE analysis compared the occulsion body and enveloped nucleocapsid proteins of EeGV and TnGV, and the polypeptide patterns also showed little similarity between these viruses. These analyses, as well as comparison of our results to those reported for other granuloviruses, indicate that EeGV represents a new granulovirus isolate. PMID- 10843835 TI - Characterization of a single-nucleocapsid nucleopolyhedrovirus of Thysanoplusia orichalcea L. (Lepidoptera: Noctuidae) from Indonesia. AB - A single-nucleocapsid nucleopolyhedrovirus (NPV) isolated from Thysanoplusia orichalcea L. (Lepidoptera:Noctuidae) (ThorNPV) in Indonesia has tetrahedral occlusion bodies (OBs) with a width of 1. 22 microm (range = 0.803-1.931 microm). The length of the virion with an envelope averaged 0.29 and 0.23 microm without an envelope. ThorNPV was propagated in Pseudoplusia includens (Walker) and its authenticity was confirmed by sequence analysis of the polyhedrin gene of the ThorNPV produced in T. orichalcea and P. includens. Polyhedrin amino acid sequence analysis revealed that ThorNPV belongs to Group II of baculoviruses and is closely related to Trichoplusia ni single nucleocapsid NPV, sharing 97.6% sequence identity. Infectivity of ThorNPV against third instar P. includens was low, with a LD(50) value of 65,636 OBs/larva. Electron microscopy of infected tissues showed many polyhedra without virions embedded, which might explain the low virulence against P. includens. Differences in virion occlusion rates between individual cells in the same tissue suggested that the inoculum consisted of at least two variants that differed in the gene(s) controlling virion occlusion. In a host range test using the LD(50) value to P. includens against Spodoptera exigua, S. frugiperda, S. eridania, Anticarsia gemmatalis, Helicoverpa zea, Trichoplusia ni, and P. includens, P. includens was the only species infected. The virus infected primarily the fat body, tracheal epithelium, and hypodermis. The genomic size of the ThorNPV is 135 kb. PMID- 10843836 TI - Effect of Bacillus thuringiensis toxins on the midgut of the nun moth Lymantria monacha. AB - Three steps of the proposed mode of action of Bacillus thuringiensis toxins have been studied in Lymantria monacha. We demonstrated that only the toxins that caused typical pathological changes in midgut epithelial cells and bound to the midgut brush border membrane were able to drastically reduce the midgut transepithelial voltage of the nun moth. PMID- 10843837 TI - Coelomomyces stegomyiae infection in adult female Aedes aegypti following the first, second, and third host blood meals. PMID- 10843838 TI - Persistence of infection in mosquitoes injected with bacteria. PMID- 10843839 TI - Isolating a species of entomophthorales using resting spore-bearing soil. PMID- 10843840 TI - Extensive methylation of a part of the CpG island located 3.0-4.5 kbp upstream to the chicken alpha-globin gene cluster may contribute to silencing the globin genes in non-erythroid cells. AB - Here, we show that in the chicken genome, the domain of alpha-globin genes is preceded by a CpG island of which the downstream part ( approximately 0.65 kbp) is heavily methylated in lymphoid cells; it is either non-methylated or undermethylated in erythroid cells. Recombinant plasmids were constructed with the corresponding DNA fragment (called "uCpG") placed upstream to a reporter CAT gene expressed from the promoter of the alpha(D) chicken globin gene. Selective methylation of CpG dinucleotides within the uCpG fragment suppressed fivefold the expression of the CAT gene, when neither this gene itself nor the alpha(D) promoter were methylated. Methylation of CpG dinucleotides within the alpha(D) gene promoter did not modify the suppression effect exerted by methylated uCpG. We interpret these results within the frame of the hypothesis postulating, that methylation of the upstream CpG island of the chicken alpha-globin gene domain may play an essential role in silencing the alpha-globin genes in non-erythroid cells. PMID- 10843841 TI - An immuno-electron microscopical analysis of transcribing multinucleosomal templates: what happens to the histones? AB - Immuno-electron microscopy was used to visualize the structure of reconstituted chromatin after in vitro transcription by purified T7 RNA polymerase. T7 RNA polymerase disrupts the nucleosomal structure in the transcribed region. This disruption is not influenced by the template, linear or supercoiled, and the presence or absence of nucleosomal positioning sequences in the transcribed region. In this study, we used monoclonal autoantibodies reacting with the nucleosome core particles and epitopes within several regions of the four different core histones. Some of the residues recognized by the autoantibodies are accessible on the surface of the nucleosomes and some are more internal and therefore less exposed at the surface. We show that the loss of the nucleosomal configuration during transcription is due to the loss of histone/DNA binding and that at least part of the histones are transferred to the nascent RNA chains. Consequently, after in vitro transcription by T7 RNA polymerase, the nucleosomal template does not conserve its original configuration, and no interaction of antigen/antibodies is observed anymore in the region that has been transcribed. Therefore, we conclude that in our in vitro transcription assay, nucleosomes are detached from the template, and not simply unfolded with histones remaining attached to the DNA. PMID- 10843842 TI - The G+C-rich discriminator region of the tyrT promoter antagonises the formation of stable preinitiation complexes. AB - RNA polymerase forms a highly stable preinitiation complex at many prokaryotic promoters in the absence of ribonucleotides. These are often characterised by the longevity of the DNA strand-separated (open) complex in the presence of heparin. In contrast, such complexes are notoriously unstable at the promoters for rRNA and tRNA under similar conditions. The high G+C content within the DNA-melting region of these promoters has been implicated in this seemingly anomalous behaviour. Here, we used rapid-pulse UV laser photo-footprinting to monitor the transient structural intermediates formed at the Escherichia coli tyrT promoter. Promoter derivatives with A+T for G/C base substitutions within the G+C-rich discriminator region (-7 to -1) augmented the stability of complexes on both linear fragments and supercoiled plasmid DNA. Analysis of the lifetime of the preinitiation complexes as a function of the discriminator sequence reveals a direct relationship between the A+T content of the DNA-melting region and the stability of the ensuing complex. Our results are consistent with the premise that a G/C block to DNA-untwisting and/or DNA-melting operates to prevent the formation of the stable isomers that are implicated in most other transcription initiation pathways. PMID- 10843843 TI - Length of CTG.CAG repeats determines the influence of mismatch repair on genetic instability. AB - We showed previously that mutations in methyl-directed mismatch repair of Escherichia coli reduced the occurrence of large deletions in (CTG.CAG)(175) repeats contained on plasmids. By contrast, other workers reported that mutations in mismatch repair increase the frequency of small-length changes in the shorter (CTG.CAG)(64). Using plasmids with a variety of lengths and purity of (CTG.CAG) repeats, we have resolved these apparently conflicting observations. We show that all lengths of (CTG.CAG) repeats are subject to small-length changes (eight repeats) in (CTG.CAG)(n) occur more readily in cells with active mismatch repair. The frequency of large deletions is proportional to the tract length; in our assays they become prominent in tracts greater than 100 repeats. Interruptions in repeat purity enhance the occurrence of large deletions. In addition, we observed a high level of incidence of deletions in (CTG.CAG) repeats for cultures passing repeatedly through stationary phase during long-term growth experiments of all strains (i.e. with active or inactive mismatch repair). These results agree with current theories on mismatch repair acting on DNA slippage events that occur in DNA triplet-repeats. PMID- 10843844 TI - The role of weak protein-protein interactions in multivalent lectin-carbohydrate binding: crystal structure of cross-linked FRIL. AB - Binding of multivalent glycoconjugates by lectins often leads to the formation of cross-linked complexes. Type I cross-links, which are one-dimensional, are formed by a divalent lectin and a divalent glycoconjugate. Type II cross-links, which are two or three-dimensional, occur when a lectin or glycoconjugate has a valence greater than two. Type II complexes are a source of additional specificity, since homogeneous type II complexes are formed in the presence of mixtures of lectins and glycoconjugates. This additional specificity is thought to become important when a lectin interacts with clusters of glycoconjugates, e.g. as is present on the cell surface. The cryst1al structure of the Glc/Man binding legume lectin FRIL in complex with a trisaccharide provides a molecular snapshot of how weak protein-protein interactions, which are not observed in solution, can become important when a cross-linked complex is formed. In solution, FRIL is a divalent dimer, but in the crystal FRIL forms a tetramer, which allows for the formation of an intricate type II cross-linked complex with the divalent trisaccharide. The dependence on weak protein-protein interactions can ensure that a specific type II cross-linked complex and its associated specificity can occur only under stringent conditions, which explains why lectins are often found forming higher order oligomers. PMID- 10843845 TI - Mechanism of hyaluronan binding and degradation: structure of Streptococcus pneumoniae hyaluronate lyase in complex with hyaluronic acid disaccharide at 1.7 A resolution. AB - Hyaluronic acid (HA) is an important constituent of the extracellular matrix; its bacterial degradation has been postulated to contribute to the spread of certain streptococci through tissue. Pneumococci and other streptococci produce hyaluronate lyase, an enzyme which depolymerizes HA, thus hyaluronate lyase might contribute directly to bacterial invasion. Although two different mechanisms for lyase action have been proposed, there was no crystallographic evidence to support those mechanisms. Here, we report the high-resolution crystal structure of Streptococcus pneumoniae hyaluronate lyase in the presence of HA disaccharide product, which ultimately provides the first crystallographic evidence for the binding of HA to hyaluronate lyase. This structural complex revealed a key interaction between the Streptococcus peneumoniae hyaluronate lyase protein and the product, and supports our previously proposed novel catalytic mechanism for HA degradation based on the native Streptococcus peneumoniae hyaluronate lyase structure. The information provided by this complex structure will likely be useful in the development of antimicrobial pharmaceutical agents. PMID- 10843846 TI - Estimating the number of protein folds and families from complete genome data. AB - Using the data on proteins encoded in complete genomes, combined with a rigorous theory of the sampling process, we estimate the total number of protein folds and families, as well as the number of folds and families in each genome. The total number of folds in globular, water- soluble proteins is estimated at about 1000, with structural information currently available for about one-third of the number. The sequenced genomes of unicellular organisms encode from approximately 25%, for the minimal genomes of the Mycoplasmas, to 70-80% for larger genomes, such as Escherichia coli and yeast, of the total number of folds. The number of protein families with significant sequence conservation was estimated to be between 4000 and 7000, with structures available for about 20% of these. PMID- 10843847 TI - A DNA structural atlas for Escherichia coli. AB - We have performed a computational analysis of DNA structural features in 18 fully sequenced prokaryotic genomes using models for DNA curvature, DNA flexibility, and DNA stability. The structural values that are computed for the Escherichia coli chromosome are significantly different from (and generally more extreme than) that expected from the nucleotide composition. To aid this analysis, we have constructed tools that plot structural measures for all positions in a long DNA sequence (e.g. an entire chromosome) in the form of color-coded wheels (http://www.cbs.dtu. dk/services/GenomeAtlas/). We find that these "structural atlases" are useful for the discovery of interesting features that may then be investigated in more depth using statistical methods. From investigation of the E. coli structural atlas, we discovered a genome-wide trend, where an extended region encompassing the terminus displays a high of level curvature, a low level of flexibility, and a low degree of helix stability. The same situation is found in the distantly related Gram-positive bacterium Bacillus subtilis, suggesting that the phenomenon is biologically relevant. Based on a search for long DNA segments where all the independent structural measures agree, we have found a set of 20 regions with identical and very extreme structural properties. Due to their strong inherent curvature, we suggest that these may function as topological domain boundaries by efficiently organizing plectonemically supercoiled DNA. Interestingly, we find that in practically all the investigated eubacterial and archaeal genomes, there is a trend for promoter DNA being more curved, less flexible, and less stable than DNA in coding regions and in intergenic DNA without promoters. This trend is present regardless of the absolute levels of the structural parameters, and we suggest that this may be related to the requirement for helix unwinding during initiation of transcription, or perhaps to the previously observed location of promoters at the apex of plectonemically supercoiled DNA. We have also analyzed the structural similarities between groups of genes by clustering all RNA and protein-encoding genes in E. coli, based on the average structural parameters. We find that most ribosomal genes (protein encoding as well as rRNA genes) cluster together, and we suggest that DNA structure may play a role in the transcription of these highly expressed genes. PMID- 10843848 TI - Alu-mediated phylogenetic novelties in gene regulation and development. AB - Differential gene expression lies at the heart of biology and is responsible for all developmental processes, including the growth and differentiation of cells. Perhaps even speciation could be defined as a change in differential gene expression over evolutionary time. The present work is a phylogenetic study of four Alu elements known to have gene regulatory functions in the human. The four elements have been shown to regulate the parathyroid hormone (PTH) gene via a negative calcium-response element, the hematopoietic cell-specific FcepsilonRI gamma receptor gene via a cis-acting positive/negative regulatory element, the CNS-specific nicotinic acetylcholine receptor alpha3 gene via a cis-acting positive/negative control element, and the T-cell-specific CD8alpha gene via a complex transcriptional regulator. The four Alu elements that impact differential gene expression were found to be differentially distributed among seven primate species (human, chimpanzee, gorilla, orangutan, baboon, rhesus, and macaque) in a way that is congruent with an accepted phylogeny of these species. The results establish a link between gene regulation and the divergence of primates. This evolutionary variation in gene regulation also suggests a novel experimental system to study the very complex transcriptional regulation of gene expression, by studying side-by-side the regulation of the same gene from two primate species that differ in the cis-acting regulatory elements of the gene. PMID- 10843849 TI - Differential role of the intermolecular base-pairs G292-C(75) and G293-C(74) in the reaction catalyzed by Escherichia coli RNase P RNA. AB - We present a systematic investigation of the thermodynamic and kinetic role of the intermolecular G292-C(75 )and G293-C(74 )Watson-Crick base-pairs in the reaction catalyzed by Escherichia coli RNase P RNA. Single turnover kinetics were analyzed for wild-type RNase P RNA and two variants with a single G to C exchange (C292 or C293), either acting on wild-type precursor tRNA (ptRNA) or derivatives carrying a complementary change at the tRNA 3'-end (G(74)CA or CG(75)A). Ground state binding of tRNA was studied using three different methods, including a novel fluorescence-based assay measuring equilibrium binding. We conclude that: (1) the role of the G293-C(74 )interaction is essentially confined to Watson Crick base-pairing, with no indication for crucial tertiary contacts involving this base-pair; (2) the G293-C(74 )pair, although being as important for ptRNA ground state binding as G292-C(75), is much less crucial to catalytic performance than the G292-C(75) pair; (3) disruption of the G292-C(75 )base-pair results in preferential destabilization of enzyme transition-state complexes; and (4) the identity of the G292-C(75) pair, as part of the higher-order structural context consisting of coplanar G292-C(75)-A258 and G291-G259-A(76 )triples, contributes to high affinity binding of ptRNA and catalytic efficiency. PMID- 10843850 TI - Characterization of a conserved alpha-helical, coiled-coil motif at the C terminal domain of the ATP-dependent FtsH (HflB) protease of Escherichia coli. AB - FtsH (HflB) is an ATP-dependent protease found in prokaryotic cells, mitochondria and chloroplasts. Here, we have identified, in the carboxy-terminal region of FtsH (HfIB), a short alpha helix predicted of forming a coiled-coil, leucine zipper, structure. This region appears to be structurally conserved. The presence of the coiled-coil motif in the Escherichia coli FtsH (HflB) was demonstrated by circular dichroism and cross-linking experiments. Mutational analysis showed that three highly conserved leucine residues are essential for FtsH (HfIB) activity in vivo and in vitro. Purified proteins mutated in the conserved leucine residues, were found to be defective in the degradation of E. coli sigma(32) and the bacteriophage lambda CII proteins. In addition, the mutant proteins were defective in the binding of CII The mutations did not interfere with the ATPase activity of FtsH (HflB). Finally, the mutant proteins were found to be more sensitive to trypsin degradation than the wild-type enzyme suggesting that the alpha helical region is an important structural element of FtsH (HflB). PMID- 10843851 TI - Signals for TBP/TATA box recognition. AB - The TATA box-binding protein (TBP) recognizes its target sites (TATA boxes) by indirectly reading the DNA sequence through its conformation effects (indirect readout). Here, we explore the molecular mechanisms underlying indirect readout of TATA boxes by TBP by studying the binding of TBP to adenovirus major late promoter (AdMLP) sequence variants, including alterations inside as well as in the sequences flanking the TATA box. We measure here the dissociation kinetics of complexes of TBP with AdMLP targets and, by phase-sensitive assay, the intrinsic bending in the TATA box sequences as well as the bending of the same sequence induced by TBP binding. In these experiments we observe a correlation of the kinetic stability to sequence changes within the TATA recognition elements. Comparison of the kinetic data with structural properties of TATA boxes in known crystalline TBP/TATA box complexes reveals several "signals" for TATA box recognition, which are both on the single base-pair level, as well as larger DNA tracts within the TATA recognition element. The DNA bending induced by TBP on its binding sites is not correlated to the stability of TBP/TATA box complexes. Moreover, we observe a significant influence on the kinetic stability of alteration in the region flanking the TATA box. This effect is limited however to target sites with alternating TA sequences, whereas the AdMLP target, containing an A tract, is not influenced by these changes. PMID- 10843852 TI - Site-directed mutagenesis of the regulatory domain of Escherichia coli carbamoyl phosphate synthetase identifies crucial residues for allosteric regulation and for transduction of the regulatory signals. AB - Carbamoyl phosphate (CP), the essential precursor of pyrimidines and arginine, is made in Escherichia coli by a single carbamoyl phosphate synthetase (CPS) consisting of 41.4 and 117.7 kDa subunits, which is feed-back inhibited by UMP and activated by IMP and ornithine. The large subunit catalyzes CP synthesis from ammonia in three steps, and binds the effectors in its 15 kDa C-terminal domain. Fifteen site-directed mutations were introduced in 13 residues of this domain to investigate the mechanism of allosteric modulation by UMP and IMP. Two mutations, K993A and V994A, decreased significantly or abolished enzyme activity, apparently by interfering with the step of carbamate synthesis, and one mutation, T974A, negatively affected ornithine activation. S948A, K954A, T974A, K993A and K993W/H995A abolished or greatly hampered IMP activation and UMP inhibition as well as the binding of both effectors, monitored using photoaffinity labeling and ultracentrifugation binding assays. V994A also decreased significantly IMP and UMP binding. L990A, V991A, H995A, G997A and G1008A had more modest effects or affected more the modulation by and the binding of one than of the other nucleotide. K993W, R1020A, R1021A and K1061A were without substantial effects. The results confirm the independence of the regulatory and catalytic centers, and also confirm functional predictions based on the X-ray structure of an IMP-CPS complex. They prove that the inhibitor UMP and the activator IMP bind in the same site, and exclude that the previously observed binding of ornithine and glutamine in this site were relevant for enzyme activation. K993 and V994 appear to be involved in the transmission of the regulatory signals triggered by UMP and IMP binding. These effectors possibly change the position of K993 and V994, and alter the intermolecular contacts mediated by the regulatory domain. PMID- 10843853 TI - Compromise and accommodation in ecotin, a dimeric macromolecular inhibitor of serine proteases. AB - Ecotin is a dimeric serine protease inhibitor from Escherichia coli which binds proteases to form a hetero-tetramer with three distinct interfaces: an ecotin ecotin dimer interface, a larger primary ecotin-protease interface, and a smaller secondary ecotin-protease interface. The contributions of these interfaces to binding and inhibition are unequal. To investigate the contribution and adaptability of each interface, we have solved the structure of two mutant ecotin trypsin complexes and compared them to the structure of the previously determined wild-type ecotin-trypsin complex. Wild-type ecotin has an affinity of 1 nM for trypsin, while the optimized mutant, ecotin Y69F, D70P, which was found using phage display technologies, inhibits rat trypsin with a K(i) value of 0.08 nM. Ecotin 67-70A, M84R which has four alanine substitutions in the ecotin-trypsin secondary binding site, along with the M84R mutation at the primary site, has a K(i) value against rat trypsin of 0.2 nM. The structure of the ecotin Y69F, D70P trypsin complex shows minor structural changes in the ecotin-trypsin tetramer. The structure of the ecotin 67-70A, M84R mutant bound to trypsin shows large deviations in the tertiary and quaternary structure of the complex. The trypsin structure shows no significant changes, but the conformation of several loop regions of ecotin are altered, resulting in the secondary site releasing its hold on trypsin. The structure of several regions previously considered to be rigid is also significantly modified. The inherent flexibility of ecotin allows it to accommodate these mutations and still maintain tight binding through the compromises of the protein-protein interfaces in the ecotin-trypsin tetramer. A comparison with two recently described ecotin-like genes from other bacteria suggests that these structural and functional features are conserved in otherwise distant bacterial lineages. PMID- 10843854 TI - Crystal structure of Pseudomonas aeruginosa PAK pilin suggests a main-chain dominated mode of receptor binding. AB - Fibers of pilin monomers (pili) form the dominant adhesin of Pseudomonas aeruginosa, and they play an important role in infections by this opportunistic bacterial pathogen. Blocking adhesion is therefore a target for vaccine development. The receptor-binding site is located in a C-terminal disulphide bonded loop of each pilin monomer, but functional binding sites are displayed only at the tip of the pilus. A factor complicating vaccination is that different bacterial strains produce distinct, and sometimes highly divergent, pilin variants. It is surprising that all strains still appear to bind a common receptor, asialo-GM1. Here, we present the 1.63 A crystal structure of pilin from P. aeruginosa strain PAK. The structure shows that the proposed receptor-binding site is formed by two beta-turns that create a surface dominated by main-chain atoms. Receptor specificity could therefore be maintained, whilst allowing side chain variation, if the main-chain conformation is conserved. The location of the binding site relative to the proposed packing of the pilus fiber raises new issues and suggests that the current fiber model may have to be reconsidered. Finally, the structure of the C-terminal disulphide-bonded loop will provide the template for the structure-based design of a consensus sequence vaccine. PMID- 10843855 TI - The 1.7 A crystal structure of BPI: a study of how two dissimilar amino acid sequences can adopt the same fold. AB - We have extended the resolution of the crystal structure of human bactericidal/permeability-increasing protein (BPI) to 1.7 A. BPI has two domains with the same fold, but with little sequence similarity. To understand the similarity in structure of the two domains, we compare the corresponding residue positions in the two domains by the method of 3D-1D profiles. A 3D-1D profile is a string formed by assigning each position in the 3D structure to one of 18 environment classes. The environment classes are defined by the local secondary structure, the area of the residue which is buried from solvent, and the fraction of the area buried by polar atoms. A structural alignment between the two BPI domains was used to compare the 3D-1D environments of structurally equivalent positions. Greater than 31% of the aligned positions have conserved 3D-1D environments, but only 13% have conserved residue identities. Analysis of the 3D 1D environmentally conserved positions helps to identify pairs of residues likely to be important in conserving the fold, regardless of the residue similarity. We find examples of 3D-1D environmentally conserved positions with dissimilar residues which nevertheless play similar structural roles. To generalize our findings, we analyzed four other proteins with similar structures yet dissimilar sequences. Together, these examples show that aligned pairs of dissimilar residues often share similar structural roles, stabilizing dissimilar sequences in the same fold. PMID- 10843856 TI - Crystal structure of dodecameric vanadium-dependent bromoperoxidase from the red algae Corallina officinalis. AB - The three-dimensional structure of the vanadium bromoperoxidase protein from the marine red macroalgae Corallina officinalis has been determined by single isomorphous replacement at 2.3 A resolution. The enzyme subunit is made up of 595 amino acid residues folded into a single alpha+beta domain. There are 12 bromoperoxidase subunits, arranged with 23-point group symmetry. A cavity is formed by the N terminus of each subunit in the centre of the dodecamer. The subunit fold and dimer organisation of the Cor. officinalis vanadium bromoperoxidase are similar to those of the dimeric enzyme from the brown algae Ascophyllum nodosum, with which it shares 33 % sequence identity. The different oligomeric state of the two algal enzymes seems to reflect separate mechanisms of adaptation to harsh environmental conditions and/or to chemically active substrates and products. The residues involved in the vanadate binding are conserved between the two algal bromoperoxidases and the vanadium chloroperoxidase from the fungus Curvularia inaequalis. However, most of the other residues forming the active-site cavity are different in the three enzymes, which reflects differences in the substrate specificity and stereoselectivity of the reaction. A dimer of the Cor. officinalis enzyme partially superimposes with the two-domain monomer of the fungal enzyme. PMID- 10843857 TI - An intermediate step in the recognition of tRNA(Asp) by aspartyl-tRNA synthetase. AB - The crystal structures of aspartyl-tRNA synthetase (AspRS) from Thermus thermophilus, a prokaryotic class IIb enzyme, complexed with tRNA(Asp) from either T. thermophilus or Escherichia coli reveal a potential intermediate of the recognition process. The tRNA is positioned on the enzyme such that it cannot be aminoacylated but adopts an overall conformation similar to that observed in active complexes. While the anticodon loop binds to the N-terminal domain of the enzyme in a manner similar to that of the related active complexes, its aminoacyl acceptor arm remains at the entrance of the active site, stabilized in its intermediate conformational state by non-specific interactions with the insertion and catalytic domains. The thermophilic nature of the enzyme, which manifests itself in a very low kinetic efficiency at 17 degrees C, the temperature at which the crystals were grown, is in agreement with the relative stability of this non productive conformational state. Based on these data, a pathway for tRNA binding and recognition is proposed. PMID- 10843858 TI - The highly refined solution structure of the cytotoxic ribonuclease alpha-sarcin reveals the structural requirements for substrate recognition and ribonucleolytic activity. AB - alpha-Sarcin selectively cleaves a single phosphodiester bond in a universally conserved sequence of the major rRNA, that inactivates the ribosome. The elucidation of the three-dimensional solution structure of this 150 residue enzyme is a crucial step towards understanding alpha-sarcin's conformational stability, ribonucleolytic activity, and its exceptionally high level of specificity. Here, the solution structure has been determined on the basis of 2658 conformationally relevant distances restraints (including stereoespecific assignments) and 119 torsional angular restraints, by nuclear magnetic resonance spectroscopy methods. A total of 60 converged structures have been computed using the program DYANA. The 47 best DYANA structures, following restrained energy minimization by GROMOS, represent the solution structure of alpha-sarcin. The resulting average pairwise root-mean-square-deviation is 0.86 A for backbone atoms and 1.47 A for all heavy atoms. When the more variable regions are excluded from the analysis, the pairwise root-mean-square deviation drops to 0.50 A and 1.00 A, for backbone and heavy atoms, respectively. The alpha-sarcin structure is similar to that reported for restrictocin, although some differences are clearly evident, especially in the loop regions. The average rmsd between the structurally aligned backbones of the 47 final alpha-sarcin structures and the crystal structure of restrictocin is 1.46 A. On the basis of a docking model constructed with alpha-sarcin solution structure and the crystal structure of a 29-nt RNA containing the sarcin/ricin domain, the regions in the protein that could interact specifically with the substrate have been identified. The structural elements that account for the specificity of RNA recognition are located in two separate regions of the protein. One is composed by residues 51 to 55 and loop 5, and the other region, located more than 11 A away in the structure, is the positively charged segment formed by residues 110 to 114. PMID- 10843859 TI - The anatomy of protein beta-sheet topology. AB - Here, we present a systematic analysis of the open-faced beta-sheet topologies in a set of non-redundant protein domain structures; in particular, we focus on the topological diversity of four-stranded beta-sheet motifs. Of the 96 topologies that are possible for a four-stranded beta-sheet, 42 were identified in known protein structures. Of these, four account for 50% of the structures that we have studied. Two sets of the topologies that were not observed may represent the section of the topological space that is not readily accessible to proteins on either thermodynamic or kinetic grounds. The first set contains topologies with alternating parallel and antiparallel beta-ladders. Their rare occurrence reflects the expectation that it is energetically unfavorable to match different hydrogen bonding patterns. The polypeptide chains in the second set of topologies go through convoluted paths and are expected to experience great kinetic frustrations during the folding processes. A knowledge of the potential causes for the topological preference of small beta-sheets also helps us to understand the topological properties of larger beta-sheet structures which frequently contain four-stranded motifs. The notion that protein topologies can only be taken from a confined and discrete space has important implications for structural genomics. PMID- 10843860 TI - pH-dependent interactions and the stability and folding kinetics of the N terminal domain of L9. Electrostatic interactions are only weakly formed in the transition state for folding. AB - The role of electrostatic interactions in the stability and the folding of the N terminal domain of the ribosomal protein L9 (NTL9) was investigated by determining the effects of varying the pH conditions. Urea denaturations and thermal unfolding experiments were used to measure the free energy of folding, DeltaG degrees, at 18 different pH values, ranging from pH 1.1 to pH 10.5. Folding rates were measured at 19 pH values between pH 2.1 and pH 9.5, and unfolding rates were determined at 15 pH values in this range using stopped-flow fluorescence experiments. The protein is maximally stable between pH 5.5 and 7.5 with a value of DeltaG degrees =4.45 kcal mol(-1). The folding rate reaches a maximum at pH 5.5, however the change in folding rates with pH is relatively modest. Over the pH range of 2.1 to 5.5 there is a small increase in folding rates, ln (k(f)) changes from 5.1 to 6.8. However, the change in stability is more dramatic, with a difference of 2.6 kcal mol(-1) between pH 2.0 and pH 5.4. The change in stability is largely due to the smaller barrier for unfolding at low pH values. The natural log of the unfolding rates varies by approximately four units between pH 2.1 and pH 5.5. The stability of the protein decreases above pH 7.5 and again the change is largely due to changes in the unfolding rate. ln (k(f)) varies by less than one unit between pH 5.5 and pH 9.5 while DeltaG degrees decreases by 2.4 kcal mol(-1) over the range of pH 5. 4 to pH 10.0, which corresponds to a change in ln K(eq) of 4.0. These studies show that pH-dependent interactions contribute significantly to the overall stability of the protein but have only a small effect upon the folding kinetics, indicating that electrostatic interactions are weakly formed in the transition state for folding. PMID- 10843861 TI - Core structure of the outer membrane lipoprotein from Escherichia coli at 1.9 A resolution. AB - The outer membrane lipoprotein of the Escherichia coli cell envelope has characteristic lipid modifications at an amino-terminal cysteine and can exist in a form bound covalently to the peptidoglycan through a carboxyl-terminal lysine. The 56-residue polypeptide moiety of the lipoprotein, designated Lpp-56, folds into a stable, trimeric helical structure in aqueous solution. The 1.9 A resolution crystal structure of Lpp-56 comprises a parallel three-stranded coiled coil including a novel alanine-zipper unit and two helix-capping motifs. The amino-terminal motif forms a hydrogen-bonding network anchoring an umbrella shaped fold. The carboxyl-terminal motif uses puckering of the tyrosine side chains as a unique docking arrangement in helix termination. The structure provides an explanation for assembly and insertion of the lipoprotein molecules into the outer membrane of gram-negative bacteria and suggests a molecular target for antibacterial drug discovery. PMID- 10843862 TI - The structure of a LysM domain from E. coli membrane-bound lytic murein transglycosylase D (MltD). AB - The LysM domain is a widespread protein module. It was originally identified in enzymes that degrade bacterial cell walls but is also present in many other bacterial proteins. Several proteins that contain the domain, such as Staphylococcal IgG binding proteins and Escherichia coli intimin, are involved in bacterial pathogenesis. LysM domains are also found in some eukaryotic proteins, apparently as a result of horizontal gene transfer from bacteria. The available evidence suggests that the LysM domain is a general peptidoglycan-binding module. We have determined the structure of this domain from E. coli membrane-bound lytic murein transglycosylase D. The LysM domain has a betaalphaalphabeta secondary structure with the two helices packing onto the same side of an anti- parallel beta sheet. The structure shows no similarity to other bacterial cell surface domains. A potential binding site in a shallow groove on surface of the protein has been identified. PMID- 10843863 TI - A minimum folding unit in the ankyrin repeat protein p16(INK4). AB - The ankyrin repeat is an abundant, 33 residue sequence motif that forms a consecutive beta-hairpin-helix-loop-helix (beta(2)alpha(2)) fold. Most ankyrin repeat proteins consist of four or more complete repeats, which provide stabilizing interactions between adjacent modules. The cyclin-dependent kinase inhibitor and tumor suppressor p16(INK4) (p16) is one of the smallest ankyrin repeat proteins with a known structure. It consists of four complete repeats plus short N and C-terminal flanking regions that are unstructured in solution. On the basis of preliminary proteolysis studies and predictions using a computer algorithm for identifying autonomous folding units, we have identified a fragment consisting of the third and fourth ankyrin repeats of p16, called p16C, that can fold independently, without the rest of the protein. Far-UV circular dichroism studies showed that p16C has a significant level of alpha-helical secondary structure, and two proline substitutions that disrupt the alpha-helical secondary structure in wild-type p16 disrupt the secondary structure in p16C. The thermal denaturation of p16C is cooperative and reversible, with a midpoint of transition at 30. 5(+/-1) degrees C. From urea-induced denaturation studies, the free energy of unfolding for p16C was estimated to be 1.7(+/-0.3) kcal/mol at 20 degrees C. (1)H-(15)N 2D NMR studies suggest that the ankyrin repeats in p16C are likely to fold into a structure similar to that of full-length p16. In order to define the minimum autonomous folding unit in p16, we have further dissected p16C into two complementary peptides, each containing a single ankyrin repeat. These peptides are unstructured in solution. Thus, p16C is the smallest ankyrin repeat module that is known to fold independently and, in general, we believe that the two ankyrin repeat fold could be the minimum structural unit for all ankyrin repeat proteins. We further discuss the significance of p16C in protein folding and engineering. PMID- 10843864 TI - Fluorescence energy transfer indicates similar transient and equilibrium intermediates in staphylococcal nuclease folding. AB - Fluorescence resonance energy transfer (FRET) is one of the few methods available to measure the rate at which a folding protein collapses. Using staphylococcal nuclease in which a cysteine residue was engineered in place of Lys64, permitted FRET measurements of the distance between the donor tryptophan 140 and 5-[[2 [(iodoacetyl)-amino]ethyl]amino]naphthalene-1-sulfonic acid-labeled Cys64. These measurements were undertaken on both equilibrium partially folded intermediates at low pH (A states), as well as transient intermediates during stopped-flow refolding. The results indicate that there is an initial collapse of the protein in the deadtime of the stopped-flow instrument, corresponding to a regain of approximately 60% of the native signal, followed by three slower transients. This is in contrast to circular dichroism measurements which show only 20-25% regain of the native secondary structure in the burst phase. Thus hydrophobic collapse precedes the formation of substantial secondary structure. The first two detected transient intermediate species have FRET properties essentially identical with those of the previously characterized equilibrium A state intermediates, suggesting similar structures between the equilibrium and transient intermediates. The effects of anions on the folding of acid-unfolded staphylococcal nuclease, and urea on the unfolding of the resulting A states, indicates that in folding the protein becomes compact prior to formation of major secondary structure, whereas in unfolding the protein expands prior to major loss of secondary structure. Comparison of the kinetics of refolding of staphylococcal nuclease, monitored by FRET, and for a proline-free variant, indicate that folding occurs via two partially folded intermediates leading to a native-like species with one (or more) proline residues in a non-native conformation. For the A states an excellent correlation between compactness measured by FRET, and compactness determined from small-angle X-ray scattering, was observed. Further, a linear relationship between compactness and free energy of unfolding was noted. Formation of soluble aggregates of the A states led to dramatic enhancement of the FRET, consistent with intermolecular fluorescence energy transfer. PMID- 10843865 TI - Conservation of substructures in proteins: interfaces of secondary structural elements in proteasomal subunits. AB - It is observed that during divergent evolution of two proteins with a common phylogenetic origin, the structural similarity of their backbones is often preserved even when the sequence similarity between them decreases to a virtually undetectable level. Here we analyzed, whether the conservation of structure along evolution involves also the local atomic structures in the interfaces between secondary structural elements. We have used as study case one protein family, the proteasomal subunits, for which 17 crystal structures are known. These include 14 different subunits of Saccharomyces cerevisiae, 2 subunits of Thermoplasma acidophilum and one subunit of Escherichia coli. The structural core of the 17 proteasomal subunits has 23 secondary structural elements. Any two adjacent secondary structural elements form a molecular interface consisting of two molecular patches. We found 61 interfaces that occurred in all 17 subunits. The 3D shape of equivalent molecular patches from different proteasomal subunits were compared by superposition. Our results demonstrate that pairs of equivalent molecular patches show an RMSD which is lower than that of randomly chosen patches from unrelated proteins. This is true even when patch comparisons with identical residues were excluded from the analysis. Furthermore it is known that the sequential dissimilarity is correlated to the RMSD between the backbones of the members of protein families. The question arises whether this is also true for local atomic structures. The results show that the correlation of individual patch RMSD values and local sequence dissimilarities is low and has a wide range from 0 to 0.41, however, it is surprising that there is a good correlation between the average RMSD of all corresponding patches and the global sequence dissimilarity. This average patch RMSD correlates slightly stronger than the C(alpha)-trace RMSD to the global sequence dissimilarity. PMID- 10843866 TI - Prologue: low-molecular-weight GTPases in the heart and circulation PMID- 10843867 TI - beta(1)-integrin and PI 3-kinase regulate RhoA-dependent activation of skeletal alpha-actin promoter in myoblasts. AB - RhoA GTPase, a regulator of actin cytoskeleton, is also involved in regulating c fos gene expression through its effect on serum response factor (SRF) transcriptional activity. We have also shown that RhoA plays a critical role in myogenesis and regulates expression of SRF-dependent muscle genes, including skeletal alpha-actin. In the present study, we examined whether the RhoA signaling pathway cross talks with other myogenic signaling pathways to modulate skeletal alpha-actin promoter activity in myoblasts. We found that extracellular matrix proteins and the beta(1)-integrin stimulated RhoA-dependent activation of the alpha-actin promoter. The muscle-specific isoform beta(1D) selectively activated the alpha-actin promoter in concert with RhoA but inhibited the c-fos promoter. In addition, focal adhesion kinase (FAK) and phosphatidylinositol (PI) 3-kinase were required for full activation of the alpha-actin promoter by RhoA. Expression of a dominant negative mutant of FAK, application of wortmannin to cultured myoblasts, or expression of a dominant negative mutant of PI 3-kinase inhibited alpha-actin promoter activity induced by RhoA. These results suggest that RhoA, beta(1)-integrin, FAK, and PI 3-kinase serve together as an important signaling network in regulating muscle gene expression. PMID- 10843868 TI - Gene transfer of dominant negative Rho kinase suppresses neointimal formation after balloon injury in pigs. AB - Restenosis after angioplasty still remains a major problem for which neointimal formation appears to play an important role. Recent studies in vitro suggested that Rho kinase, a target protein of Rho, is important in various cellular functions. We thus examined whether Rho kinase is involved in the restenotic changes after balloon injury. In vivo gene transfer was performed immediately after balloon injury in both sides of the porcine femoral arteries with adenoviral vector encoding either a dominant negative form of Rho kinase (AdDNRhoK) or beta-galactosidase (AdLacZ) as a control. One week after the transfer, immunohistochemistry confirmed the successful gene expression in the vessel wall, whereas 2 wk after the transfer, Western blotting showed the functional upregulation of Rho kinase at the AdLacZ site and its suppression at the AdDNRhoK site. Angiography showed the development of a stenotic lesion at the AdLacZ site where histological neointimal formation was noted, whereas those changes were significantly suppressed at the AdDNRhoK site. These results indicate that Rho kinase is involved in the pathogenesis of neointimal formation after balloon injury in vivo. PMID- 10843869 TI - P2Y(1), P2Y(2), P2Y(4), and P2Y(6) receptors are coupled to Rho and Rho kinase activation in vascular myocytes. AB - In the cardiovascular system, activation of ionotropic (P2X receptors) and metabotropic (P2Y receptors) P2 nucleotide receptors exerts potent and various responses including vasodilation, vasoconstriction, and vascular smooth muscle cell proliferation. Here we examined the involvement of the small GTPase RhoA in P2Y receptor-mediated effects in vascular myocytes. Stimulation of cultured aortic myocytes with P2Y receptor agonists induced an increase in the amount of membrane-bound RhoA and stimulated actin cytoskeleton organization. P2Y receptor agonist-induced actin stress fiber formation was inhibited by C3 exoenzyme and the Rho kinase inhibitor Y-27632. Stimulation of actin cytoskeleton organization by extracellular nucleotides was also abolished in aortic myocytes expressing a dominant negative form of RhoA. Extracellular nucleotides induced contraction and Y-27632-sensitive Ca(2+) sensitization in aortic rings. Transfection of Swiss 3T3 cells with P2Y receptors showed that Rho kinase-dependent actin stress fiber organization was induced in cells expressing P2Y(1), P2Y(2), P2Y(4), or P2Y(6) receptor subtypes. Our data demonstrate that P2Y(1), P2Y(2), P2Y(4), and P2Y(6) receptor subtypes are coupled to activation of RhoA and subsequently to Rho dependent signaling pathways. PMID- 10843870 TI - Inhibition of Rho protein stimulates iNOS expression in rat vascular smooth muscle cells. AB - Inducible nitric oxide synthase (iNOS) in vascular smooth muscle cells (VSMCs) is upregulated in arterial injury and plays a role in regulating VSMC proliferation and restenosis. Inflammatory cytokines [e.g., interleukin-1beta (IL-1beta)] released during vascular injury induce iNOS. Small GTP-binding proteins of the Ras superfamily play a major role in IL-1beta-dependent signaling pathways. In this study, we examined the role of Rho GTPases in regulating iNOS expression in VSMCs. Treatment of VSMCs with mevastatin, which inhibits isoprenylation of Rho and other small GTP-binding proteins, produced significantly higher amounts of IL 1beta-evoked NO and iNOS protein compared with control. Similarly, bacterial toxins [Toxin B from Clostridium difficile and C3 ADP-ribosyl transferase (C3) toxin from Clostridium botulinium] that specifically inactivate Rho proteins increased NOS products (NO and citrulline) and iNOS expression. Toxin B increased the activity of iNOS promoter-reporter construct in VSMCs. Both toxins enhanced IL-1beta-stimulated iNOS expression and NO production. These data demonstrate for the first time that inhibition of Rho induces iNOS and suggest a role for Rho protein in IL-1beta-stimulated NO production in VSMCs. PMID- 10843871 TI - The Rho effector, PKN, regulates ANF gene transcription in cardiomyocytes through a serum response element. AB - The low-molecular-weight GTP-binding protein RhoA mediates hypertrophic growth and atrial natriuretic factor (ANF) gene expression in neonatal rat ventricular myocytes. Neither the effector nor the promoter elements through which Rho exerts its regulatory effects on ANF gene expression have been elucidated. When constitutively activated forms of Rho kinase and two protein kinase C-related kinases, PKN (PRK1) and PRK2, were compared, only PKN generated a robust stimulation of a luciferase reporter gene driven by a 638-bp fragment on the ANF promoter. This ANF promoter fragment contains a proximal serum response element (SRE) and an Sp-1-like element required for the transcriptional response to phenylephrine (PE). This response was inhibited by dominant negative Rho. The ability of dominant negative Rho to inhibit the response to PE and the ability of PKN to stimulate ANF reporter gene expression were both lost when the SRE was mutated. Mutation of the Sp-1-like element also attenuated the response to PKN. A minimal promoter driven by ANF SRE sequences was sufficient to confer Rho- and PKN-mediated gene expression. Interestingly, PKN preferentially stimulated the ANF versus the c-fos SRE reporter gene. Thus PKN and Rho are able to regulate transcriptional activation of the ANF SRE by a common element that could implicate PKN as a downstream effector of Rho in transcriptional responses associated with hypertrophy. PMID- 10843872 TI - NTS A(2a) purinoceptor activation elicits hindlimb vasodilation primarily via a beta-adrenergic mechanism. AB - Previously, we have shown that activation of adenosine A(2a) receptors in the subpostremal nucleus tractus solitarii (NTS) via microinjection of the selective A(2a) receptor agonist CGS-21680 elicits potent, dose-dependent decreases in mean arterial pressure and preferential, marked hindlimb vasodilation. Although A(2a) receptor activation does not change lumbar sympathetic nerve activity, it does markedly enhance the preganglionic adrenal sympathetic nerve activity, which will increase epinephrine release and could subsequently elicit hindlimb vasodilation via activation of beta(2)-adrenergic receptors. Therefore we investigated whether this hindlimb vasodilation was due to neural or humoral mechanisms. In chloralose urethan-anesthetized male Sprague-Dawley rats, we monitored cardiovascular responses to stimulation of NTS adenosine A(2a) receptors (CGS-21680, 20 pmol/50 nl) in the intact control animals; after pretreatment with propranolol (2 mg/kg iv), a beta-adrenergic antagonist; after bilateral lumbar sympathectomy; after bilateral adrenalectomy; and after combined bilateral lumbar sympathectomy and adrenalectomy. After beta-adrenergic blockade, stimulation of NTS adenosine A(2a) receptors produced a pressor response and a hindlimb vasoconstriction. Lumbar sympathectomy reduced the vasodilation seen in the intact animals by approximately 40%, and adrenalectomy reduced it by approximately 80%. The combined sympathectomy and adrenalectomy virtually abolished the hindlimb vasodilation evoked by NTS A(2a) receptor activation. We conclude that the preferential, marked hindlimb vasodilation produced by stimulation of NTS adenosine A(2a) receptors is mediated by both the efferent sympathetic nerves directed to the hindlimb and the adrenal glands via primarily a beta-adrenergic mechanism. PMID- 10843873 TI - Inhaled NO reduces leukocyte-endothelial cell interactions and myocardial dysfunction in endotoxemic rats. AB - Inhaled nitric oxide (NO) has been shown to have some protective effect in the peripheral distal inflamed vasculature. The objective of the study was to determine whether inhaled NO would reduce endotoxin-induced leukocyte activation and myocardial contractile dysfunction. Rats were treated with either saline or endotoxin (10 mg/kg iv) and then allowed to breathe (4 h) either air or air plus NO (10 ppm). In endotoxemic rats, mesenteric venular endothelium leukocyte firm adhesion increased compared with control rats (1.15 +/- 0.32 vs. 4.08 +/- 0.96 leukocytes/100 microm; P < 0.05). Inhaled NO significantly attenuated endotoxin induced venular endothelium leukocyte adhesion (4.08 +/- 0.96 vs. 1.86 +/- 0.76 leukocytes/100 microm; P < 0.05) and FITC-conjugated anti-intercellular adhesion molecule-1 fluorescence intensity. Endotoxin-induced myocardial dysfunction and leukocyte content increases were reduced in inhaled NO-treated rats. These observations suggest that inhaled NO reduces the degree of cardiovascular dysfunction and inflammation in endotoxemic rats. PMID- 10843874 TI - Ischemic preconditioning prevents I/R-induced alterations in SR calcium calmodulin protein kinase II. AB - Although Ca(2+)/calmodulin-dependent protein kinase II (CaMK II) is known to modulate the function of cardiac sarcoplasmic reticulum (SR) under physiological conditions, the status of SR CaMK II in ischemic preconditioning (IP) of the heart is not known. IP was induced by subjecting the isolated perfused rat hearts to three cycles of brief ischemia-reperfusion (I/R; 5 min ischemia and 5 min reperfusion), whereas the control hearts were perfused for 30 min with oxygenated medium. Sustained I/R in control and IP groups was induced by 30 min of global ischemia followed by 30 min of reperfusion. The left ventricular developed pressure, rate of the left ventricular pressure, as well as SR Ca(2+)-uptake activity and SR Ca(2+)-pump ATPase activity were depressed in the control I/R hearts; these changes were prevented upon subjecting the hearts to IP. The beneficial effects of IP on the I/R-induced changes in contractile activity and SR Ca(2+) pump were lost upon treating the hearts with KN-93, a specific CaMK II inhibitor. IP also prevented the I/R-induced depression in Ca(2+)/calmodulin dependent SR Ca(2+)-uptake activity and the I/R-induced decrease in the SR CaMK II activity; these effects of IP were blocked by KN-93. The results indicate that IP may prevent the I/R-induced alterations in SR Ca(2+) handling abilities by preserving the SR CaMK II activity, and it is suggested that CaMK II may play a role in mediating the beneficial effects of IP on heart function. PMID- 10843875 TI - Impaired nitric oxide-mediated vasodilation in transgenic sickle mouse. AB - Transgenic sickle mice expressing human beta(S)- and beta(S-Antilles)-globins show intravascular sickling, red blood cell adhesion, and attenuated arteriolar constriction in response to oxygen. We hypothesize that these abnormalities and the likely endothelial damage, also reported in sickle cell anemia, alter nitric oxide (NO)-mediated microvascular responses and hemodynamics in this mouse model. Transgenic mice showed a lower mean arterial pressure (MAP) compared with control groups (90 +/- 7 vs. 113 +/- 8 mmHg, P < 0.00001), accompanied by increased endothelial nitric oxide synthase (eNOS) expression. N(G)-nitro-L-arginine methyl ester (L-NAME), a nonselective inhibitor of NOS, caused an approximately 30% increase in MAP and approximately 40% decrease in the diameters of cremaster muscle arterioles (branching orders: A2 and A3) in both control and transgenic mice, confirming NOS activity; these changes were reversible after L-arginine administration. Aminoguanidine, an inhibitor of inducible NOS, had no effect. Transgenic mice showed a decreased (P < 0.02-0.01) arteriolar dilation in response to NO-mediated vasodilators, i.e., ACh and sodium nitroprusside (SNP). Indomethacin did not alter the responses to ACh and SNP. Forskolin, a cAMP activating agent, caused a comparable dilation of A2 and A3 vessels ( approximately 44 and 70%) in both groups of mice. Thus in transgenic mice, an increased eNOS/NO activity results in lower blood pressure and diminished arteriolar responses to NO-mediated vasodilators. Although the increased NOS/NO activity may compensate for flow abnormalities, it may also cause pathophysiological alterations in vascular tone. PMID- 10843876 TI - G(i)-dependent suppression of beta(1)-adrenoceptor effects in ventricular myocytes from NE-treated guinea pigs. AB - It has been suggested that there is a preferential coupling in heart muscle between the inhibitory G protein (G(i)) and the beta(2)-subtype of the beta adrenergic receptor (beta-AR), since pertussis toxin (which inactivates G(i)) reveals latent beta(2)-ARs in rat and mouse myocytes. We have previously shown that guinea pigs treated with norepinephrine (NE) for 7 days have myocytes that are desensitized to beta-AR-agonist stimulation, and that pertussis toxin restores these responses. The purpose of the present investigation was to determine whether pertussis toxin specifically upregulated beta(2)-ARs in myocytes from NE-treated guinea pigs. The sole beta-AR subtype in control guinea pig myocytes was confirmed as beta(1)-AR by radioligand binding, single-cell autoradiography, and concentration-response curves to isoproterenol in contracting myocytes. In contrast, a minor pool of beta(2)-ARs was observed in rat myocytes by use of the same methods. NE treatment decreased the maximum isoproterenol response (relative to high Ca(2+)) from 0.89 +/- 0.06 to 0.58 +/- 0.08 (n = 7, P < 0.01) and the pD(2) (-log EC(50)) from 8.8 +/- 0.2 to 7.5 +/- 0.2 (n = 7, P < 0.01). Pertussis toxin treatment increased the isoproterenol-to Ca(2+) ratio to 0.88 +/- 0.04 (n = 6, P < 0.05) and the pD(2) to 8.6 +/- 0.3 (P < 0.01). This was not mediated by increases in either number or function of beta(2) ARs. G(i) is therefore able to modulate beta(1)-AR responses in guinea pig myocytes. PMID- 10843877 TI - Somatostatin receptor subtype expression and function in human vascular tissue. AB - In animal models the somatostatin analog angiopeptin inhibits intimal hyperplasia by acting primarily through somatostatin receptor 2 (SSTR-2). However, the results of clinical trials using angiopeptin have been disappointing. In this study we showed that human blood vessels express high levels of SSTR-1 with significantly lower levels of SSTR-2 and -4. Samples of normal veins and arteries, as well as atherosclerotic arteries, expressed predominantly SSTR-1. In addition, the levels of SSTR-1 varied between individuals, indicating that the vascular disease process may have affected SSTR gene expression. Immunocytochemical studies demonstrated that SSTR-1 was present in endothelial but not vascular smooth muscle cells. No evidence of SSTR-3 or -5 expression was detected in normal or diseased blood vessels. Two endothelial cell preparations, ECV304 and human umbilical vein endothelial cells, were investigated and shown to express only SSTR-1 and -4. Exposure of these cells to 10 nM somatostatin or 10 nM SSTR-1-specific agonist resulted in alterations to the actin cytoskeleton, as characterized by a loss of actin stress fibers coupled with an increase in lamellipodia formation at the plasma membrane. These results suggest that the lack of effectiveness of angiopeptin in humans may be due to the differential expression of SSTR-1 by human endothelial cells. PMID- 10843878 TI - Essential roles of IkappaB kinases alpha and beta in serum- and IL-1-induced human VSMC proliferation. AB - Interleukin-1 (IL-1) is a potent vascular smooth muscle cell (VSMC) mitogen, which can stimulate cells via activation of nuclear factor-kappaB (NF-kappaB) following phosphorylation of its inhibitory subunit (IkappaB). Because the proliferative effect of IL-1 is additive with that of serum, the present studies assessed the role of IkappaB kinases (IKKs) and NF-kappaB in both IL-1- and serum induced VSMC proliferation. IL-1beta (1 ng/ml) induced marked and persistent NF kappaB activation in VSMC that was maximal at 1 h and persisted for 3 days. There was a 3-fold increase in DNA synthesis after acute IL-1 exposure (24-96 h) and a 12-fold increase after chronic IL-1 exposure (>7 days). Electrophoretic mobility shift assay and supershift analysis indicated that IL-1-induced NF-kappaB complexes consisted of p65/p50 heterodimers and p50 homodimers. Human saphenous vein smooth muscle cells (HSVSMC) were transiently cotransfected with expression plasmids encoding a dominant negative mutant form of either IKKalpha or IKKbeta, in which K(44) was mutated to A (K44A), and a green fluorescent protein expression plasmid that allows identification of transfected cells. IL-1 induced nuclear localization of p65 in 95% of cells transfected with vector alone but in only 69% and 26% of cells expressing IKKalpha (K44A) or IKKbeta (K44A), respectively. Likewise, proliferation increased 3.2-fold in IL-1-treated HSVSMC which had been transfected with vector alone, but only 2.2- and 1.5-fold proliferation in HSVSMC expressing IKKalpha (K44A) or IKKbeta (K44A), respectively. Although serum activated NF-kappaB transiently, serum-induced proliferation was markedly attenuated in HSVSMC expressing IKKalpha (K44A) and IKKbeta (K44A) compared with HSVSMC transfected with vector alone. The results support an essential role of IKKs in the proliferative response of HSVSMC to IL-1 and to serum. PMID- 10843879 TI - Role of EDHF in conduction of vasodilation along hamster cheek pouch arterioles in vivo. AB - We tested whether local and conducted responses to ACh depend on factors released from endothelial cells (EC) in cheek pouch arterioles of anesthetized hamsters. ACh was delivered from a micropipette (1 s, 500 nA), while arteriolar diameter (rest, approximately 40 microm) was monitored at the site of application (local) and at 520 and 1,040 microm upstream (conducted). Under control conditions, ACh elicited local (22-65 microm) and conducted (14-44 microm) vasodilation. Indomethacin (10 microM) had no effect, whereas N(omega)-nitro-L-arginine (100 microM) reduced local and conducted vasodilation by 5-8% (P < 0.05). Miconazole (10 microM) or 17-octadecynoic acid (17-ODYA; 10 microM) diminished local vasodilation by 15-20% and conducted responses by 50-70% (P < 0.05), suggesting a role for cytochrome P-450 (CYP) metabolites in arteriolar responses to ACh. Membrane potential (E(m)) was recorded in smooth muscle cells (SMC) and in EC identified with dye labeling. At rest (control E(m), typically -30 mV), ACh evoked local (15-32 mV) and conducted (6-31 mV) hyperpolarizations in SMC and EC. Miconazole inhibited SMC and EC hyperpolarization, whereas 17-ODYA inhibited hyperpolarization of SMC but not of EC. Findings indicate that ACh-induced release of CYP metabolites from arteriolar EC evoke SMC hyperpolarization that contributes substantively to conducted vasodilation. PMID- 10843880 TI - Decreased passive stiffness of cardiac myocytes and cardiac tissue from copper deficient rat hearts. AB - Passive stiffness characteristics of isolated cardiac myocytes, papillary muscles, and aortic strips from male Holtzman rats fed a copper-deficient diet for approximately 5 wk were compared with those of rats fed a copper-adequate diet to determine whether alterations in these characteristics might accompany the well-documented cardiac hypertrophy and high incidence of ventricular rupture characteristic of copper deficiency. Stiffness of isolated cardiac myocytes was assessed from measurements of cellular dimensional changes to varied osmotic conditions. Stiffness of papillary muscles and aortic strips was determined from resting length-tension analyses and included steady-state characteristics, dynamic viscoelastic stiffness properties, and maximum tensile strength. The primary findings were that copper deficiency resulted in cardiac hypertrophy with increased cardiac myocyte size and fragility, decreased cardiac myocyte stiffness, and decreased papillary muscle passive stiffness, dynamic stiffness, and tensile strength and no alteration in aortic connective tissue passive stiffness or tensile strength. These findings suggest that a reduction of cardiac myocyte stiffness and increased cellular fragility could contribute to the reduced overall cardiac tissue stiffness and the high incidence of ventricular aneurysm observed in copper-deficient rats. PMID- 10843881 TI - Spontaneous fluctuations in cerebral blood flow: insights from extended-duration recordings in humans. AB - To determine the dependence of cerebral blood flow (CBF) on arterial pressure over prolonged time periods, we measured beat-to-beat changes in mean CBF velocity in the middle cerebral artery (transcranial Doppler) and mean arterial pressure (Finapres) continuously for 2 h in six healthy subjects (5 men and 1 woman, 18-40 yr old) during supine rest. Fluctuations in velocity and pressure were quantified by the range [(peak - trough)/mean] and coefficients of variation (SD/mean) in the time domain and by spectral analysis in the frequency domain. Mean velocity and pressure over the 2-h recordings were 60 +/- 7 cm/s and 83 +/- 8 mmHg, associated with ranges of 77 +/- 8 and 89 +/- 10% and coefficients of variation of 9.3 +/- 2.2 and 7.9 +/- 2.3%, respectively. Spectral power of the velocity and pressure was predominantly distributed in the frequency range of 0.00014-0.1 Hz and increased inversely with frequency, indicating characteristics of an inverse power law (1/f(alpha)). However, linear regression on a log-log scale revealed that the slope of spectral power of pressure and velocity was steeper in the high-frequency (0.02-0.5 Hz) than in the low-frequency range (0.002-0.02 Hz), suggesting different regulatory mechanisms in these two frequency ranges. Furthermore, the spectral slope of pressure was significantly steeper than that of velocity in the low-frequency range, consistent with the low transfer function gain and low coherence estimated at these frequencies. We conclude that 1) long-term fluctuations in CBF velocity are prominent and similar to those observed in arterial pressure, 2) spectral power of CBF velocity reveals characteristics of 1/f(alpha), and 3) cerebral attenuation of oscillations in CBF velocity in response to changes in pressure may be more effective at low than that at high frequencies, emphasizing the frequency dependence of cerebral autoregulation. PMID- 10843882 TI - Endoplasmic reticulum contribution to the relaxant effect of cGMP- and cAMP elevating agents in feline aorta. AB - The contribution of endoplasmic reticulum (ER) and phosphorylation of phospholamban (PLB) to the relaxant effect of cGMP- and cAMP-elevating agents was studied in feline aorta. Sodium nitroprusside (NP, 100 microM) completely relaxed contracture induced by 10 microM norepinephrine. This NP-induced relaxation was partially prevented by tetraethylammonium, suggesting that a fraction of NP induced relaxation was mediated by activation of K(+) channels. In the absence and presence of tetraethylammonium, the relaxant effect of NP was associated with a significant increase in Ser(16) phosphorylation of PLB immunodetected by phosphorylation site-specific antibodies. The relaxant effect of NP on aortic strips precontracted with 80 mM KCl was significantly reduced by 1 microM thapsigargin. This decrease, which represents the ER contribution to the relaxant effect of NP, reached 23 +/- 9% at 100 microM NP and was closely associated with a dose-dependent increase in Ser(16) phosphorylation (128 +/- 49% over control at 100 microM NP). Effects of NP were associated with a significant increase in activity of protein kinase G and were mimicked by 8-bromo-cGMP. Forskolin produced a dose-dependent relaxant effect on KCl-induced contracture, which reached 64 +/- 8% at 50 microM and was associated with an increase in phosphorylation of Ser(16) residue of PLB (88 +/- 18% over control). Thapsigargin reduced this relaxant effect by 38 +/- 9%. 8-Bromo-cAMP mimicked effects of forskolin. The ER-mediated relaxant effect and the increase in Ser(16) phosphorylation produced by forskolin were partially blocked by the protein kinase A inhibitor H-89 (5 microM). The results indicate that ER partially contributes to the relaxant effect of NP and forskolin in feline aorta. This effect may be mediated by the associated increase in Ser(16) phosphorylation of PLB. PMID- 10843883 TI - Structural and functional remodeling of skeletal muscle microvasculature is induced by simulated microgravity. AB - Hindlimb unloading of rats results in a diminished ability of skeletal muscle arterioles to constrict in vitro and elevate vascular resistance in vivo. The purpose of the present study was to determine whether alterations in the mechanical environment (i.e., reduced fluid pressure and blood flow) of the vasculature in hindlimb skeletal muscles from 2-wk hindlimb-unloaded (HU) rats induces a structural remodeling of arterial microvessels that may account for these observations. Transverse cross sections were used to determine media cross sectional area (CSA), wall thickness, outer perimeter, number of media nuclei, and vessel luminal diameter of feed arteries and first-order (1A) arterioles from soleus and the superficial portion of gastrocnemius muscles. Endothelium dependent dilation (ACh) was also determined. Media CSA of resistance arteries was diminished by hindlimb unloading as a result of decreased media thickness (gastrocnemius muscle) or reduced vessel diameter (soleus muscle). ACh-induced dilation was diminished by 2 wk of hindlimb unloading in soleus 1A arterioles, but not in gastrocnemius 1A arterioles. These results indicate that structural remodeling and functional adaptations of the arterial microvasculature occur in skeletal muscles of the HU rat; the data suggest that these alterations may be induced by reductions in transmural pressure (gastrocnemius muscle) and wall shear stress (soleus muscle). PMID- 10843884 TI - Metabolism of VLDL is increased in streptozotocin-induced diabetic rat hearts. AB - In streptozotocin (STZ)-induced diabetic rats, we previously showed an increased heparin-releasable (luminal) lipoprotein lipase (LPL) activity from perfused hearts. To study the effect of this enlarged LPL pool on triglyceride (TG)-rich lipoproteins, we examined the metabolism of very-low-density lipoprotein (VLDL) perfused through control and diabetic hearts. Diabetic rats had elevated TG levels compared with control. However, fasting for 16 h abolished this difference. When the plasma lipoprotein fraction of density <1.006 g/ml from fasted control and diabetic rats was incubated in vitro with purified bovine or rat LPL, VLDL from diabetic animals was hydrolyzed as proficiently as VLDL from control animals. Post-heparin plasma lipolytic activity was comparable in control and diabetic animals. However, perfusion of control and diabetic rats with heparinase indicated that diabetic hearts had larger amounts of LPL bound to heparan sulfate proteoglycan-binding sites. [(3)H]VLDL obtained from control rats, when recirculated through the isolated heart, disappeared at a significantly faster rate from diabetic than from control rat hearts. This increased VLDL-TG hydrolysis was essentially abolished by prior perfusion of the diabetic heart with heparin, implicating LPL in this process. These findings suggest that the enlarged LPL pool in the diabetic heart is present at a functionally relevant location (at the capillary lumen) and is capable of hydrolyzing VLDL. This could increase the delivery of free fatty acid to the heart, and the resultant metabolic changes could induce the subsequent cardiomyopathy that is observed in the chronic diabetic rat. PMID- 10843885 TI - Peroxynitrite reversibly inhibits Ca(2+)-activated K(+) channels in rat cerebral artery smooth muscle cells. AB - Peroxynitrite (ONOO(-)) is a contractile agonist of rat middle cerebral arteries. To determine the mechanism responsible for this component of ONOO(-) bioactivity, the present study examined the effect of ONOO(-) on ionic current and channel activity in rat cerebral arteries. Whole cell recordings of voltage-clamped cells were made under conditions designed to optimize K(+) current. The effects of iberiotoxin, a selective inhibitor of large-conductance Ca(2+)-activated K(+) (BK) channels, and ONOO(-) (10-100 microM) were determined. At a pipette potential of +50 mV, ONOO(-) inhibited 39% of iberiotoxin-sensitive current. ONOO(-) was selective for iberiotoxin-sensitive current, whereas decomposed ONOO( ) had no effect. In excised, inside-out membrane patches, channel activity was recorded using symmetrical K(+) solutions. Unitary currents were sensitive to increases in internal Ca(2+) concentration, consistent with activity due to BK channels. Internal ONOO(-) dose dependently inhibited channel activity by decreasing open probability and mean open times. The inhibitory effect of ONOO(-) could be overcome by reduced glutathione. Glutathione, added after ONOO(-), restored whole cell current amplitude to control levels and reverted single channel gating to control behavior. The inhibitory effect of ONOO(-) on membrane K(+) current is consistent with its contractile effects in isolated cerebral arteries and single myocytes. Taken together, our data suggest that ONOO(-) has the potential to alter cerebral vascular tone by inhibiting BK channel activity. PMID- 10843886 TI - Inducible polymorphic ventricular tachyarrhythmias in a transgenic mouse model with a long Q-T phenotype. AB - We created a mouse model with a prolonged Q-T interval and spontaneous arrhythmias by overexpressing the NH(2) terminus and first transmembrane segment (Kv1.1N206Tag) of a delayed rectifier potassium channel (LQT(+/-) mouse). Analyses were performed using whole cell recordings of cardiac myocytes, surface electrocardiography, and programmed electrical stimulation. Action potential duration (APD) was prolonged to the same extent and was more highly variable in myocytes derived from LQT(+/-) and LQT(+/+) mice than in myocytes derived from wild-type (WT) FVB mice. Under ketamine anesthesia, the Q-T interval of both LQT(+/+) and LQT(+/-) mice was comparably prolonged versus that of WT mice. Stimulation of the right ventricle using an intracardiac catheter induced polymorphic ventricular tachyarrhythmias in 50% of the LQT(+/-) mice and 36% of the LQT(+/+) mice, whereas polymorphic ventricular tachyarrhythmias were not inducible in WT mice. The analyses of LQT(+/-) and LQT(+/+) mice indicate that prolongation of the Q-T interval in LQT mice is associated with prolonged APD, increased dispersion of APD among cardiocytes, and inducibility of polymorphic ventricular tachycardia, providing the substrate for spontaneous arrhythmias in these animals. PMID- 10843887 TI - Evidence for modulation of smooth muscle force by the p38 MAP kinase/HSP27 pathway. AB - Mitogen-activated protein (MAP) kinases signal to proteins that could modify smooth muscle contraction. Caldesmon is a substrate for extracellular signal related kinases (ERK) and p38 MAP kinases in vitro and has been suggested to modulate actin-myosin interaction and contraction. Heat shock protein 27 (HSP27) is downstream of p38 MAP kinases presumably participating in the sustained phase of muscle contraction. We tested the role of caldesmon and HSP27 phosphorylation in the contractile response of vascular smooth muscle by using inhibitors of both MAP kinase pathways. In intact smooth muscle, PD-098059 abolished endothelin-1 (ET-1)-stimulated phosphorylation of ERK MAP kinases and caldesmon, but p38 MAP kinase activation and contractile response remained unaffected. SB-203580 reduced muscle contraction and inhibited p38 MAP kinase and HSP27 phosphorylation but had no effect on ERK MAP kinase and caldesmon phosphorylation. In permeabilized muscle fibers, SB-203580 and a polyclonal anti-HSP27 antibody attenuated ET-1 dependent contraction, whereas PD-098059 had no effect. These results suggest that ERK MAP kinases phosphorylate caldesmon in vivo but that activation of this pathway is unnecessary for force development. The generation of maximal force may be modulated by the p38 MAP kinase/HSP27 pathway. PMID- 10843888 TI - Differential expression of KvLQT1 isoforms across the human ventricular wall. AB - Long Q-T mutant (KvLQT1) K(+) channels associate with their regulatory subunit IsK to produce the slow component of the delayed rectifier potassium (I(Ks)) cardiac current. The amplitude of KvLQT1 current depends on the expression of a KvLQT1 splice variant (isoform 2) that exerts strong dominant negative effects on the full-length KvLQT1 protein (isoform 1). We used RNase protection assays to determine the relative expression of KvLQT1 isoforms 1 and 2 and IsK mRNAs in human ventricular layers. Overall expression of KvLQT1 and IsK genes was similar in the three layers. However, there was a significant difference in the ratio between KvLQT1 isoforms 1 and 2. Isoform 2 represented 25.2 +/- 2.3%, 31.7 +/- 1.2%, and 24.9 +/- 1.7% of total KvLQT1 expression in left ventricular endocardial, midmyocardial, and epicardial tissues, respectively. Similar data were obtained from right ventricular samples. COS-7 cells were intranuclearly injected with KvLQT1 isoforms 1 or 2 plus IsK cDNAs, using two different isoform 2-to-isoform 1 ratios. Cells injected with an isoform 2-to-isoform 1 ratio mimicking that in the midmyocardium showed a K(+) current with approximately 75% reduced amplitude compared with those injected with a ratio mimicking that in the epicardium. Our results suggest that differential expression of KvLQT1 isoform 2 in endocardial, midmyocardial, and epicardial tissues is responsible for differential I(Ks) amplitude and contributes to the regional action potential heterogeneity observed across the ventricular wall. PMID- 10843889 TI - Remote arteriolar dilations in response to muscle contraction under capillaries. AB - In hamster cremaster muscle, it has been shown previously that contraction of skeletal muscle fibers underlying small groups of capillaries (modules) induces dilations that are proportional to metabolic rate in the two arteriolar generations upstream of the stimulated capillaries (Berg BR, Cohen KD, and Sarelius IH. Am J Physiol Heart Circ Physiol 272: H2693-H2700, 1997). These remote dilations were hypothesized to be transmitted via gap junctions and not perivascular nerves. In the present study, halothane (0.07%) blocked dilation in the module inflow arteriole, and dilation in the second arteriolar generation upstream, the branch arteriole, was blocked by both 600 mosM sucrose and halothane but not tetrodotoxin (2 microM). Dilations in both arterioles were not blocked by the gap junction uncoupler 18-beta-glycyrrhetinic acid (40 microM), and 80 mM KCl did not block dilation of the module inflow arteriole. These data implicate a gap junctional-mediated pathway insensitive to 18-beta-glycyrrhetinic acid in dilating the two arterioles upstream of the capillary module during "remote" muscle contraction. Dilation in the branch arteriole, but not the module inflow arteriole, was attenuated by 100 microM N(omega)-nitro-L-arginine. Thus selective contraction of muscle fibers underneath capillaries results in dilations in the upstream arterioles that have characteristics consistent with a signal that is transmitted along the vessel wall through gap junctions, i.e., a conducted vasodilation. The observed insensitivities to 18-beta-glycyrrhetinic acid, to KCl, and to N(omega)-nitro-L-arginine suggest, however, that there are multiple signaling pathways by which remote dilations can be initiated in these microvessels. PMID- 10843890 TI - SERCA2a activity correlates with the force-frequency relationship in human myocardium. AB - The present investigation addresses whether protein expression and function of sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA2a) and phospholamban (PLB) correlate in failing and nonfailing human myocardium. SERCA2a activity and protein expression, PLB phosphorylation, and the force-frequency relationship (FFR) have been determined in right atrium (RA) and left ventricle (LV) from nonfailing (NF, n = 12) and terminally failing [dilated cardiomyopathy (DCM), n = 12] human hearts. Only in LV of DCM hearts was SERCA2a activity significantly decreased [maximal turnover rate (V(max)) = 196 +/- 11 and 396 +/- 30 nmol. mg( 1). min(-1) in LV and RA, respectively], whereas protein expression of SERCA2a in the different chambers was unchanged in NF (3.9 +/- 0.3 and 3.2 +/- 0.4 densitometric units in LV and RA, respectively) and DCM hearts (4.8 +/- 0.8 and 3.4 +/- 0.1 densitometric units in LV and RA, respectively). Phosphorylation of PLB was higher in LV than in RA in NF (Ser(16): 180.5 +/- 19.0 vs. 56.8 +/- 6.0 densitometric units; Thr(17): 174.6 +/- 11.2 vs. 37.4 +/- 8.9 densitometric units) and DCM hearts (Ser(16): 132.0 +/- 5.4 vs. 22.4 +/- 3.5 densitometric units; Thr(17): 131.2 +/- 10.9 vs. 9.2 +/- 2.4 densitometric units). SERCA2a function, but not protein expression, correlated well with the functional parameters of the FFR in DCM and NF human hearts. Regulation of SERCA2a function depends on the phosphorylation of PLB at Ser(16) and Thr(17). However, direct SERCA2a regulation might also be affected by an unknown mechanism. PMID- 10843891 TI - Selective plasmalogen substrate utilization by thrombin-stimulated Ca(2+) independent PLA(2) in cardiomyocytes. AB - Thrombin stimulation of rabbit ventricular myocytes activates a membrane associated, Ca(2+)-independent phospholipase A(2) (PLA(2)) capable of hydrolyzing plasmenylcholine (choline plasmalogen), plasmanylcholine (alkylacyl choline phospholipid), and phosphatidylcholine substrates. To identify the endogenous phospholipid substrates, we quantified the effects of thrombin stimulation on diradyl phospholipid mass and arachidonic acid and lysophospholipid production. Thrombin stimulation resulted in a selective decrease in arachidonylated plasmenylcholine, with no change in arachidonylated phosphatidylcholine. The decrease in arachidonylated plasmenylcholine was accompanied by an increase in plasmenylcholine species containing linoleic and linolenic acids at the sn-2 position. A decrease in arachidonylated plasmenylethanolamine was also observed after thrombin stimulation, with no concomitant change in arachidonylated phosphatidylethanolamine. Thrombin stimulation resulted in the selective production of lysoplasmenylcholine, with no increase in lysophosphatidylcholine content. There was no evidence for significant acetylation of lysophospholipids to form platelet-activating factor. Arachidonic acid released after thrombin stimulation was rapidly oxidized to prostacyclin. Thus thrombin-stimulated Ca(2+) independent PLA(2) selectively hydrolyzes arachidonylated plasmalogen substrates, resulting in production of lysoplasmalogens and prostacyclin as the principal bioactive products. PMID- 10843892 TI - Effects of estrogen on venous function in rats with chronic heart failure. AB - The effect of 17beta-estradiol on venous function was investigated in ovariectomized rats with heart failure. Rats (50-60 days old) were ovariectomized and implanted with 60-day-release pellets that contain 17beta-estradiol (1.5 mg) or vehicle. The left coronary artery was ligated 7 days later. Another group of ovariectomized rats was given vehicle pellets and then a sham operation was performed. The rats were studied while under pentobarbital anesthesia at 7 wk after ligation. Ligated rats, relative to sham groups, had lower mean arterial pressure (MAP, -34 mmHg) and cardiac output (CO, -38%); higher arterial resistance (R(A), +12%) and venous resistance (R(V), +116%); mean circulatory filling pressure (MCFP, +40%) and left ventricular end-diastolic pressure (LVEDP, +11 mmHg); and similar cardiovascular responses to norepinephrine (NE). Treatment of ligated rats with 17beta-estradiol increased CO (+16%); reduced R(A) (-16%), R(V) (-35%), MCFP (-23%), and LVEDP (-3 mmHg); and augmented MAP, R(V,) and MCFP responses to NE. Therefore, 17beta-estradiol reduced MCFP, and this reduced preload (LVEDP). 17beta-Estradiol decreased R(V), which, along with decreased R(A) (afterload), led to an increase in CO. 17beta-Estradiol likely augmented vasoconstriction to NE through an improvement on the cardiovascular status. PMID- 10843893 TI - Cardioprotective effect of chronic hyperglycemia: effect on hypoxia-induced apoptosis and necrosis. AB - It is generally accepted that mild forms of diabetes render the heart resistant to an ischemic insult. Because myocytes incubated chronically in medium containing high concentrations of glucose (25 mM) develop into a diabetes-like phenotype, we tested the hypothesis that high-glucose treatment diminishes hypoxia-induced injury. In support of this hypothesis, we found that cardiomyocytes incubated for 3 days with medium containing 25 mM glucose showed less hypoxia-induced apoptosis and necrosis than cells exposed to medium containing 5 mM glucose (control). Indeed, whereas 27% of control cells became necrotic after 1 h of chemical hypoxia with 10 mM deoxyglucose and 5 mM amobarbital (Amytal), only 11% of the glucose-treated cells became necrotic. Similarly, glucose treatment reduced the extent of apoptosis from 32% to 12%. This beneficial effect of glucose treatment was associated with a 40% reduction in the Ca(2+) content of the hypoxic cell. Glucose treatment also mediated an upregulation of the cardioprotective factor Bcl-2 but did not affect the cellular content of the proapoptotic factors Bax and Bad. Nonetheless, the phosphorylation state of Bad was shifted in favor of its inactive, phosphorylated form after high glucose treatment. These data suggest that glucose treatment renders the cardiomyocyte resistant to hypoxia-induced apoptosis and necrosis by preventing the accumulation of Ca(2+) during hypoxia, promoting the upregulation of Bcl-2, and enhancing the inactivation of Bad. PMID- 10843894 TI - Nitric oxide modulation of TNF-alpha-induced cardiac contractile dysfunction is concentration dependent. AB - Whereas previous studies suggest that tumor necrosis factor-alpha (TNF-alpha) induces cardiac contraction-relaxation deficits, the mechanisms remain unclear. Our recent studies have implicated cardiac-derived nitric oxide (NO). This study examined the detrimental and protective effects of NO donors S-nitroso-N-acetyl penicillamine (SNAP) or (Z)-1- [N-(3-ammonio-propyl)-N-(n-propyl)amino]diazen-1 ium- 1,2diolate (PAPA/NO) on TNF-alpha-related changes in cardiac contractile function (Langendorff), cellular injury, and intracellular myocyte Ca(2+) concentration ([Ca(2+)](i)). Myocytes were incubated in the presence/absence of TNF-alpha (200-500 pg/ml x 10(5) cells) for 3 h; subsets of myocytes were incubated with one of several concentrations of SNAP or PAPA/NO (0.1, 0.3, 0.5, and 1.5 mM) for 15 min before TNF-alpha challenge. Supernatant creatine kinase (CK), cell viability (Trypan blue dye exclusion), and myocyte [Ca(2+)](i) (fura 2 acetoxymethyl ester) were measured. In parallel experiments, cardiac function (Langendorff) was examined after TNF-alpha challenge in the presence or absence of SNAP or PAPA/NO (0.1 and 1.5 mM). TNF-alpha in the absence of an NO donor impaired cardiac contraction and relaxation and produced cardiomyocyte injury. Pretreating perfused hearts or isolated cardiomyocytes with a low concentration of either SNAP or PAPA/NO decreased TNF-alpha-mediated cardiac injury and improved contractile dysfunction, whereas high concentrations of NO donor exacerbated TNF-alpha-mediated cardiac effects. These data provide one explanation for the conflicting reports of beneficial versus detrimental effects of NO in the face of inflammation and suggest that the effects of NO on organ function are concentration dependent; low concentrations of NO are cardioprotective, whereas high concentrations of NO are deleterious. PMID- 10843895 TI - Efficacy and specificity of bFGF increased collateral flow in experimental peripheral arterial insufficiency. AB - Angiogenic growth factors could prove to be useful in managing peripheral arterial insufficiency. The present study was designed to evaluate the dose response of basic fibroblast growth factor (bFGF), the efficacy of critical routes and dosing regimens, and the specificity of action in rats with peripheral arterial insufficiency. Bilateral ligation of femoral arteries greatly reduces blood flow capacity to the calf muscles but does not impair resting flow needs. Collateral blood flow to calf muscles was determined 16 days postocclusion, during treadmill running, with (85)Sr and (141)Ce microspheres, in blinded randomized trials that included intra-arterial and intravenous infusions and subcutaneous injections of recombinant human bFGF. Peak blood flow of 75-80 ml. min(-1). 100 g(-1) for calf muscle was observed at a bFGF dose of 5 microg. kg( 1). day(-1) (ia for 14 days) compared with 50 ml. min(-1). 100 g(-1) for vehicle groups. Similar increases in collateral blood flow were observed with short-term or prolonged and continuous or intermittent delivery of bFGF by any route. Collateral blood flows were similar in corresponding muscles across both limbs. Vascular remodeling induced by bFGF required attendant vascular occlusion, inasmuch as vessels in the normal nonoccluded vascular tree were unresponsive to circulating bFGF. Improvement in collateral blood flow with exogenous bFGF is robust, amenable to short-term administration, and requires vascular occlusion to be effective. PMID- 10843896 TI - Systemic and renal hemodynamics after moderate hemodilution with HbOCs in anesthetized rabbits. AB - Hb-based O(2)-carrying solutions (HbOCs) have been developed as red blood cell substitutes for use in patients undergoing hemodilution. Variously modified Hb with diverse solution properties have been shown to produce variable hemodynamic responses. We examined, through pulsed-Doppler velocimetry, the systemic and renal hemodynamic effects of dextran-benzene-tetracarboxylate-conjugated (Hb-Dex BTC), bis(3,5-dibromosalicyl)fumarate cross-linked (alphaalpha-Hb), and o raffinose-polymerized (o-raffinose-Hb) Hb perfused in rabbits after moderate hemodilution (30% hematocrit), and we compared the effects of these Hb solutions with the effects elicited by plasma volume expanders. In addition, vascular hindrance (resistance/blood viscosity at 128.5 s(-1)) was calculated to determine whether a moderate decrease in the viscosity of blood mixed with HbOCs may impair vasoconstriction as a result of autoregulation after infusion of cell-free Hb. No changes were observed in renal hemodynamics after hemodilution with reference or Hb solutions. Increase in blood pressure and vascular resistance was found with Hb-Dex-BTC and alphaalpha-Hb (for 180 min) and, to a lesser extent, with o raffinose-Hb (for 120 min). Furthermore, Hb-Dex-BTC (high viscosity) and o raffinose-Hb (medium viscosity) induced comparable increases in vascular hindrance (from 0.091 to 0. 159 and from 0.092 to 0.162 cm(-1), respectively) but far less than that produced by alphaalpha-Hb (low viscosity, from 0.092 to 0.200 cm(-1)). These results suggest that maintaining the viscosity of blood by infusing solutions with high viscosity makes it possible to limit vasoconstriction due to autoregulation mechanisms and mainly caused by hemodilution per se. PMID- 10843897 TI - Exercise training restores adenosine-induced relaxation in coronary arteries distal to chronic occlusion. AB - We previously reported that canine collateral-dependent coronary arteries exhibit impaired relaxation to adenosine but not sodium nitroprusside. In contrast, exercise training enhances adenosine sensitivity of normal porcine coronary arteries. These results stimulated the hypothesis that chronic coronary occlusion and exercise training produce differential effects on cAMP- versus cGMP-mediated relaxation. To test this hypothesis, Ameroid occluders were surgically placed around the proximal left circumflex coronary artery (LCx) of female Yucatan miniature swine 8 wk before initiating sedentary or exercise training (treadmill run, 16 wk) protocols. Relaxation to the cAMP-dependent vasodilators adenosine (10(-7) to 10(-3) M) and isoproterenol (3 x 10(-8) to 3 x 10(-5) M) were impaired in collateral-dependent LCx versus nonoccluded left anterior descending (LAD) arterial rings isolated from sedentary but not exercise-trained pigs. Furthermore, adenosine-mediated reductions in simultaneous tension and myoplasmic free Ca(2+) were impaired in LCx versus LAD arteries isolated from sedentary but not exercise-trained pigs. In contrast, relaxation in response to the cAMP dependent vasodilator forskolin (10(-9) to 10(-5) M) and the cGMP-dependent vasodilator sodium nitroprusside (10(-9) to 10(-4) M) was not different in LCx versus LAD arteries of sedentary or exercise-trained animals. These data suggest that chronic occlusion impairs receptor-dependent, cAMP-mediated relaxation; receptor-independent cAMP- and cGMP-mediated relaxation were unimpaired. Importantly, exercise training restores cAMP-mediated relaxation of collateral dependent coronary arteries. PMID- 10843898 TI - Influence of glycogen storage on vascular smooth muscle metabolism. AB - The role of glycogen as an oxidative substrate for vascular smooth muscle (VSM) remains controversial. To elucidate the importance of glycogen as an oxidative substrate and the influence of glycogen flux on VSM substrate selection, we systematically altered glycogen levels and measured metabolism of glucose, acetate, and glycogen. Hog carotid arteries with glycogen contents ranging from 1 to 11 micromol/g were isometrically contracted in physiological salt solution containing 5 mM [1-(13)C]glucose and 1 mM [1, 2-(13)C]acetate at 37 degrees C for 6 h. [1-(13)C]glucose, [1, 2-(13)C]acetate, and glycogen oxidation were simultaneously measured with the use of a (13)C-labeled isotopomer analysis of glutamate. Although oxidation of glycogen increased with the glycogen content of the tissue, glycogen oxidation contributed only approximately 10% of the substrate oxidized by VSM. Whereas [1-(13)C]glucose flux, [3-(13)C]lactate production from [1-(13)C]glucose, and [1, 2-(13)C]acetate oxidation were not regulated by glycogen content, [1-(13)C]glucose oxidation was significantly affected by the glycogen content of VSM. However, [1-(13)C]glucose remained the primary ( approximately 40-50%) contributor to substrate oxidation. Therefore, we conclude that glucose is the predominate substrate oxidized by VSM, and glycogen oxidation contributes minimally to substrate oxidation. PMID- 10843899 TI - Potassium channels modulate cerebral autoregulation during acute hypertension. AB - We tested the hypothesis that constriction of cerebral arterioles during acute increases in blood pressure is attenuated by activation of potassium (K(+)) channels. We tested the effects of inhibitors of calcium-dependent K(+) channels [iberiotoxin (50 nM) and tetraethylammonium (TEA, 1 mM)] on changes in arteriolar diameter during acute hypertension. Diameter of cerebral arterioles (baseline diameter = 46 +/- 2 microm, mean +/- SE) was measured using a cranial window in anesthetized rats. Arterial pressure was increased from a control value of 96 +/- 1 mmHg to 130, 150, 170, and 200 mmHg by intravenous infusion of phenylephrine. Increases in arterial pressure from baseline to 130 and 150 mmHg decreased the diameter of cerebral arterioles by 5-10%. Greater increases in arterial pressure produced large increases in arteriolar diameter (i.e., "breakthrough of autoregulation"). Iberiotoxin or TEA inhibited increases in arteriolar diameter when arterial pressure was increased to 170 and 200 mmHg. The change in arteriolar diameter at 200 mmHg was 20 +/- 3% and -1 +/- 4% in the absence and presence of iberiotoxin, respectively. These findings suggest that calcium dependent K(+) channels attenuate cerebral microvascular constriction during acute increases in arterial pressure, and that increases in arteriolar diameter at high levels of arterial pressure are not simply a passive phenomenon. PMID- 10843900 TI - Identification of ecto-PKC on surface of human platelets: role in maintenance of latent fibrinogen receptors. AB - Human platelets express a protein phosphorylation system on their surface. A specific protein kinase C (PKC) antibody, monoclonal antibody (MAb) 1.9, which binds to the catalytic domain of PKC and inhibits its activity, causes the aggregation of intact platelets while inhibiting the phosphorylation of platelet surface proteins. Photoaffinity labeling with 100 nM 8-azido-[alpha(32)P]ATP identified this ecto-PKC as a single surface protein of 43 kDa sensitive to proteolysis by extracellular 0.0005% trypsin. Inhibition of the binding of 8 azido-[alpha(32)P]ATP to the 43-kDa surface protein by MAb 1.9 identified this site as the active domain of ecto-PKC. Covalent binding of the azido-ATP molecule to the 43-kDa surface protein inhibited the phosphorylative activity of the platelet ecto-PKC. Furthermore, PKC pseudosubstrate inhibitory peptides directly induced the aggregation of platelets and inhibited azido-ATP binding to the 43 kDa protein. Platelet aggregation induced by MAb 1.9 and by PKC inhibitory peptides required the presence of fibrinogen and resulted in an increase in the level of intracellular free calcium concentration. This increase in intracellular free calcium concentration induced by MAb 1.9 was found to be dependent on the binding of fibrinogen to activated GPIIb/IIIa integrins, suggesting that MAb 1.9 causes Ca(2+) flux through the fibrinogen receptor complex. We conclude that a decrease in the state of phosphorylation of platelet surface proteins caused by inhibition of ecto-PKC results in membrane rearrangements that can induce the activation of latent fibrinogen receptors, leading to platelet aggregation. Accordingly, the maintenance of a physiological steady state of phosphorylation of proteins on the platelet surface by ecto-PKC activity appears to be one of the homeostatic mechanisms that maintain fibrinogen receptors of circulating platelets in a latent state that cannot bind fibrinogen. PMID- 10843901 TI - Endothelial dysfunction does not require loss of endothelial nitric oxide synthase. AB - Whereas altered nitric oxide (NO.) formation from endothelial nitric oxide synthase (NOS) causes impaired vascular reactivity in a number of cardiovascular diseases, questions remain regarding how endothelial injury results in impaired NO. formation. It is unknown if loss of NOS expression or activity is required or if other factors are involved. Detergent treatment has been used to induce endothelial dysfunction. Therefore, NOS and NO. synthesis were characterized in a rat heart model of endothelial injury and dysfunction induced by the detergent Triton X-100. Cardiac NO. formation was directly measured by electron paramagnetic resonance spectroscopy. NOS activity was determined by the L [(14)C]arginine conversion assay. Western blots and immunohistology were applied to define the amounts of NOS present in heart tissue before and after Triton treatment. Immunoelectron microscopy was performed to assess intracellular NOS distribution. A short bolus of Triton X-100, 0.25%, abolished responses to histamine and calcium ionophore while preserving response to nitroprusside. Complete blockade of NO. generation occurred after Triton treatment, but NOS activity assayed with addition of exogenous substrate and cofactors was unchanged, and identical 135-kDa NOS bands were seen on Western blots, indicating that NOS was not removed from the heart or structurally damaged by Triton. Immunohistochemistry showed no change in NOS localization after Triton treatment, and immunoelectron microscopy revealed similar NOS distribution in the plasma membrane and intracellular membranes. These results demonstrate that the endothelial dysfunction was due to decreased NO. synthesis but was not caused by loss or denaturation of NOS. Thus endothelial dysfunction due to mild endothelial membrane injury may occur in the presence of active NOS and is triggered by loss of NOS substrates or cofactors. PMID- 10843903 TI - Physiological time-series analysis using approximate entropy and sample entropy. AB - Entropy, as it relates to dynamical systems, is the rate of information production. Methods for estimation of the entropy of a system represented by a time series are not, however, well suited to analysis of the short and noisy data sets encountered in cardiovascular and other biological studies. Pincus introduced approximate entropy (ApEn), a set of measures of system complexity closely related to entropy, which is easily applied to clinical cardiovascular and other time series. ApEn statistics, however, lead to inconsistent results. We have developed a new and related complexity measure, sample entropy (SampEn), and have compared ApEn and SampEn by using them to analyze sets of random numbers with known probabilistic character. We have also evaluated cross-ApEn and cross SampEn, which use cardiovascular data sets to measure the similarity of two distinct time series. SampEn agreed with theory much more closely than ApEn over a broad range of conditions. The improved accuracy of SampEn statistics should make them useful in the study of experimental clinical cardiovascular and other biological time series. PMID- 10843902 TI - Sulfonylurea receptor ligands modulate stretch-induced ANF secretion in rat atrial myocyte culture. AB - Stretch-induced atrial natriuretic factor (ANF) secretion was studied in cultures of neonate atrial appendage myocytes. Stretch, applied for 40 min by hypotonic swelling, increased the mean area of 44 individually imaged myocytes by 4.8-8.8% (P < 0.0001) at 6 min and by 2.3-6.2% (P < 0.05) at 35 min. Stretch increased immunoreactive ANF release by 42% (P < 0.05) from a baseline of 315 pg/ml. The ATP-sensitive K(+) (K(ATP))-channel blocker tolbutamide (100 micromol/l) increased the stretch-stimulated release to 84% (P < 0.01) over baseline, whereas lower concentrations (1, 10, and 30 micromol/l) had no stimulatory effect. The K(ATP)-channel opener diazoxide (0.1, 1, 10, 30, and 100 micromol/l) inhibited stretch- plus tolbutamide-stimulated ANF release in a concentration-dependent manner, with IC(50) = 2.2 micromol/l, although 100 micromol/l diazoxide did not reduce the increase in mean cell area. The stretch-stimulated K(ATP) current, monitored in 82 whole cell recordings with sulfonylurea receptor (SUR) ligands, was inversely correlated with the stretch-induced ANF release (r(2) = 0.79, P < 0. 0001). In the absence of stretch, the K(ATP) current had no relationship with baseline ANF release, and baseline ANF release was not affected by the K(ATP) channel modulators. The results show that SUR ligands that open K(ATP) channels inhibit stretch-induced ANF release in atrial myocytes, in correlation with the stretch-activated K(ATP) current. The subcellular site of action of the SUR ligands-plasmalemma or intracellular organelles-remains to be determined. PMID- 10843904 TI - ET-1 in the myocardial interstitium: relation to myocyte ECE activity and expression. AB - Increased plasma levels of endothelin-1 (ET-1) have been identified in congestive heart failure (CHF), but local myocardial interstitial ET-1 levels and the relation to determinants of ET-1 synthesis remain to be defined. Accordingly, myocardial interstitial ET-1 levels and myocyte endothelin-converting enzyme (ECE)-1 activity and expression with the development of CHF were examined. Pigs were instrumented with a microdialysis system to measure myocardial interstitial ET-1 levels with pacing CHF (240 beats/min, 3 wk; n = 9) and in controls (n = 14). Plasma ET-1 was increased with CHF (15 +/- 1 vs. 9 +/- 1 fmol/ml, P < 0.05) as was total myocardial ET-1 content (90 +/- 15 vs. 35 +/- 5 fmol/g, P < 0.05). Paradoxically, myocardial interstitial ET-1 was decreased in CHF (32 +/- 4 vs. 21 +/- 2 fmol/ml, P < 0.05), which indicated increased ET-1 uptake by the left ventricular (LV) myocardium with CHF. In isolated LV myocyte preparations, ECE-1 activity was increased by twofold with CHF (P < 0.05). In LV myocytes, both ECE 1a and ECE-1c mRNAs were detected, and ECE-1a expression was upregulated fivefold in CHF myocytes (P < 0.05). In conclusion, this study demonstrated compartmentalization of ET-1 in the myocardial interstitium and enhanced ET-1 uptake with CHF. Thus a local ET-1 system exists at the level of the myocyte, and determinants of ET-1 biosynthesis are selectively regulated within this myocardial compartment in CHF. PMID- 10843905 TI - Differential role of ionotropic glutamatergic mechanisms in responses to NTS P(2x) and A(2a) receptor stimulation. AB - Activation of ATP P(2x) receptors in the subpostremal nucleus tractus solitarii (NTS) via microinjection of alpha,beta-methylene ATP (alpha,beta-MeATP) elicits fast initial depressor and sympathoinhibitory responses that are followed by slow, long-lasting inhibitory effects. Activation of NTS adenosine A(2a) receptors via microinjection of CGS-21680 elicits slow, long-lasting decreases in arterial pressure and renal sympathetic nerve activity (RSNA) and an increase in preganglionic adrenal sympathetic nerve activity (pre-ASNA). Both P(2x) and A(2a) receptors may operate via modulation of glutamate release from central neurons. We investigated whether intact glutamatergic transmission is necessary to mediate the responses to NTS P(2x) and A(2a) receptor stimulation. The hemodynamic and neural (RSNA and pre-ASNA) responses to microinjections of alpha,beta-MeATP (25 pmol/50 nl) and CGS-21680 (20 pmol/50 nl) were compared before and after pretreatment with kynurenate sodium (KYN; 4.4 nmol/100 nl) in chloralose-urethan anesthetized male Sprague-Dawley rats. KYN virtually abolished the fast responses to alpha,beta-MeATP and tended to enhance the slow component of the neural responses. The depressor responses to CGS-21680 were mostly preserved after pretreatment with KYN, although the increase in pre-ASNA was reduced by one-half following the glutamatergic blockade. We conclude that the fast responses to stimulation of NTS P(2x) receptors are mediated via glutamatergic ionotropic mechanisms, whereas the slow responses to stimulation of NTS P(2x) and A(2a) receptors are mediated mostly via other neuromodulatory mechanisms. PMID- 10843906 TI - Chronic hypoxia, pregnancy, and endothelium-mediated relaxation in guinea pig uterine and thoracic arteries. AB - Vasodilation that occurs during normal pregnancy is associated with enhanced relaxation and decreased contractile response to agonists, which are in part due to increased stimulated and basal nitric oxide (NO). In preeclampsia and/or pregnancies carried at high altitude (HA), this normal vascular adjustment is reversed or diminished. We previously reported that HA exposure did not inhibit the pregnancy-associated decrease in contractile response to agonist or basal NO in guinea pig uterine arteries (UA). We therefore sought to determine whether altitude interfered with effects of pregnancy on endothelium-dependent relaxation through a reduction in stimulated NO. We examined the relaxation response to ACh in UA and bradykinin in thoracic arteries (TA) and effects of NO inhibition with 200 microM N(G)-nitro-L-arginine (L-NNA) in arterial rings isolated from nonpregnant and pregnant guinea pigs exposed throughout gestation to low altitude (LA, 1,600 m, n = 26) or HA (3,962 m, n = 22). In pregnant UA, relaxation to ACh was enhanced (P < 0.05) at both altitudes and NO inhibition diminished, but did not reverse, ACh relaxation. The effect of L-NNA on the relaxation response to ACh was less in HA than in LA animals (P = 0.0021). In nonpregnant UA, relaxation to ACh was similar in LA and HA animals. L-NNA reversed the relaxation response to ACh at HA but not at LA. In TA, relaxation to bradykinin was unaltered by pregnancy or altitude and was completely reversed by NO inhibition. These data suggest that effects of NO inhibition are diminished in UA during pregnancy at HA. Additional studies are needed to confirm whether these effects are mediated through inhibition of stimulated NO. HA exposure did not inhibit relaxation to ACh, perhaps because of stimulation of other vasodilators. PMID- 10843907 TI - Changes of beta-adrenergic signaling in compensated human cardiac hypertrophy depend on the underlying disease. AB - In human heart failure, desensitization of the beta-adrenergic signal transduction has been reported to be one of the main pathophysiological alterations. However, data on the beta-adrenergic system in human compensated cardiac hypertrophy are very limited. Therefore, we studied the myocardial beta adrenergic signaling in patients suffering from hypertrophic obstructive cardiomyopathy (HOCM, n = 9) or from aortic valve stenosis (AoSt, n = 8). beta Adrenoceptor density determined by [(125)I]iodocyanopindolol binding was reduced in HOCM and AoSt compared with nonhypertrophied, nonfailing myocardium (NF) of seven organ donors. In HOCM the protein expression of stimulatory G protein alpha subunit (G(s)alpha) measured by immunoblotting was unchanged, whereas the inhibitory G protein alpha-subunit (Galpha(i-2)) was increased. In contrast, in AoSt, Galpha(i-2) protein was unchanged, but G(s)alpha protein was increased. Adenylyl cyclase stimulation by isoproterenol was reduced in HOCM but not in AoSt. Plasma catecholamine levels were normal in all patients. In conclusion, both forms of hypertrophy are associated with beta-adrenoceptor downregulation but with different changes at the G protein level that occur before symptomatic heart failure due to progressive dilatation of the left ventricle develops and are not due to elevated plasma catecholamine levels. PMID- 10843908 TI - Nonanticoagulant heparin inhibits NF-kappaB activation and attenuates myocardial reperfusion injury. AB - Heparin reduces ischemia-reperfusion injury to myocardium. This effect has been attributed to complement inhibition, but heparin also has other activities that might diminish ischemia-reperfusion. To further probe these mechanisms, we compared heparin or an o-desulfated nonanticoagulant heparin with greatly reduced anticomplement activity. When given at the time of coronary artery reperfusion in a canine model of myocardial infarction, heparin or o-desulfated heparin equally reduced neutrophil adherence to ischemic-reperfused coronary artery endothelium, influx of neutrophils into ischemic-reperfused myocardium, myocardial necrosis, and release of creatine kinase into plasma. Heparin or o-desulfated heparin also prevented dysfunction of endothelial-dependent coronary relaxation following ischemic injury. In addition, heparin and o-desulfated heparin inhibited translocation of the transcription nuclear factor-kappaB (NF-kappaB) from the cytoplasm to the nucleus in human endothelial cells and decreased NF-kappaB DNA binding in human endothelium and ischemic-reperfused rat myocardium. Thus heparin and nonanticoagulant heparin decrease ischemia-reperfusion injury by disrupting multiple levels of the inflammatory cascade, including the novel observation that heparins inhibit activation of the proinflammatory transcription factor NF kappaB. PMID- 10843909 TI - Histamine-induced depolarization: ionic mechanisms and role in sustained contraction of rabbit cerebral arteries. AB - The role of membrane depolarization in the histamine-induced contraction of the rabbit middle cerebral artery was examined by simultaneous measurements of membrane potential and isometric force. Histamine (1-100 microM) induced a concentration-dependent sustained contraction associated with sustained depolarization. Action potentials were observed during depolarization caused by histamine but not by high-K(+) solution. K(+)-induced contraction was much smaller than sustained contraction associated with the same depolarization caused by histamine. Nifedipine attenuates histamine-induced sustained contraction by 80%, with no effect on depolarization. Inhibition of nonselective cation channels with Co(2+) (100-200 microM) reversed the histamine-induced depolarization and relaxed the arteries but induced only a minor change in K(+)-induced contraction. In the presence of Co(2+) and in low-Na(+) solution, histamine-evoked depolarization and contraction were transient. We conclude that nonselective cation channels contribute to histamine-induced sustained depolarization, which stimulates Ca(2+) influx through voltage-dependent Ca(2+) channels participating in contraction. The histamine-induced depolarization, although an important and necessary mechanism, cannot fully account for sustained contraction, which may be due in part to augmentation of currents through voltage-dependent Ca(2+) channels and Ca(2+) sensitization of the contractile process. PMID- 10843911 TI - Cyclosporin A inhibits cardiac hypertrophy and enhances cardiac dysfunction during postinfarction failure in rats. AB - Calcineurin has recently been implicated as a mediator in the signaling pathways that transform intracellular calcium signals to the phenotype of myocardial hypertrophy. The present study was conducted to examine the effects of cyclosporin A (CsA), an inhibitor of calcineurin, on myocardial hypertrophy and remodeling during congestive heart failure (CHF) in rats. After ligation of the left coronary artery, rats were randomized to treatment with CsA or vehicle for 14 days. Compared with vehicle, CsA substantially attenuated myocardial hypertrophy in the CHF rats as assessed by alterations in ventricular weight-to tibial length ratios (P < 0.05). Myocardial gene expression of skeletal alpha actin was also reduced in the failing left ventricle (LV) after treatment with CsA (P < 0. 05), although the mRNA levels were still substantially elevated relative to those of sham rats. CsA-induced inhibition of compensatory myocardial hypertrophy in the CHF rats led to increased dilatation of the LV cavity and reduced fractional shortening and peak positive and negative first derivatives of LV pressure (P < 0. 05). Plasma renin and endothelin-1 levels were increased in the CHF-CsA rats, providing humoral cues of aggravated cardiac function. Thus this study supports a crucial role of calcineurin-dependent pathways in the mechanisms of compensatory myocardial hypertrophy during CHF. In addition, our data indicate that inhibition of compensatory myocardial hypertrophy exerts detrimental effects on cardiac remodeling and function after myocardial infarction. PMID- 10843910 TI - Role of intracellular Ca(2+) release in histamine-induced depolarization in rabbit middle cerebral artery. AB - The role of Ca(2+) mobilization from intracellular stores and Ca(2+)-activated Cl(-) channels in caffeine- and histamine-induced depolarization and contraction of the rabbit middle cerebral artery has been studied by recording membrane potential and isometric force. Caffeine induced a transient contraction and a transient followed by sustained depolarization. The transient depolarization was abolished by ryanodine, DIDS, and niflumic acid, suggesting involvement of Ca(2+) activated Cl(-) channels. Histamine-evoked transient contraction in Ca(2+)-free solution was abolished by ryanodine or by caffeine-induced depletion of Ca(2+) stores. Ryanodine slowed the development of depolarization induced by histamine in Ca(2+)-containing solution but did not affect its magnitude. In arteries treated with 1 mM Co(2+), histamine elicited a transient depolarization and contraction, which was abolished by ryanodine. DIDS and niflumic acid reduced histamine-evoked depolarization and contraction. Histamine caused a sustained depolarization and contraction in low-Cl(-) solution. These results suggest that Ca(2+) mobilization from ryanodine-sensitive stores is involved in histamine induced initial, but not sustained, depolarization and contraction. Ca(2+) activated Cl(-) channels contribute mainly to histamine-induced initial depolarization and less importantly to sustained depolarization, which is most likely dependent on activation of nonselective cation channels. PMID- 10843912 TI - Nicotine increases ventricular vulnerability to fibrillation in hearts with healed myocardial infarction. AB - The vulnerability of the infarcted hearts to ventricular fibrillation (VF) was tested in in situ canine hearts during nicotine infusion. The activation pattern was mapped with 477 bipolar electrodes in open-chest anesthetized dogs (n = 8) 5 6 wk after permanent occlusion of the left anterior descending coronary artery. Nicotine (129 +/- 76 ng/ml) lengthened (P < 0.01) the pacing cycle length at which VF was induced from 171 +/- 8.9 to 210 +/- 14. 7 ms. Nicotine selectively amplified the magnitude of conduction time and monophasic action potential (MAP) amplitude and duration (MAPA and MAPD, respectively) alternans in the epicardial border zone (EBZ) but not in the normal zone. With critical reduction of the MAPA and MAPD in the EBZ, conduction block occurred across the long axis of the EBZ cells. Block led immediately to reentry formation in the EBZ with a mean period of 105 +/- 10 ms, which, after one to two rotations, degenerated to VF. Nicotine widened the range of diastolic intervals over which the dynamic MAPD restitution curve had a slope >1. We conclude that nicotine facilitates conduction block, reentry, and VF in hearts with healed myocardial infarction by increasing the magnitude of depolarization and repolarization alternans consistent with the restitution hypothesis of vulnerability to VF. PMID- 10843913 TI - Regulation of types I and III NOS in ovine uterine arteries by daily and acute estrogen exposure. AB - Nitric oxide contributes to estrogen-mediated uterine vasodilation; however, the nitric oxide synthases (NOS) involved and their location within uterine arteries are incompletely documented. We investigated the effects of repetitive daily and acute estradiol-17beta (E(2)beta) exposure on uterine hemodynamics and NOS abundance and localization in uterine arteries from nonpregnant ovariectomized ewes receiving daily intravenous E(2)beta (1 microg/kg, n = 5) or no E(2)beta (n = 7) for 5 days to determine NOS abundance, cGMP contents, and NOS immunohistochemistry. Daily E(2)beta increased basal and E(2)beta-mediated rises in uterine blood flow (UBF) 36 and 43% (<0.01), respectively, calcium-dependent NOS activity 150% (P < 0.02) in endothelium-intact and -denuded ( approximately 40% of total NOS) arteries, and cGMP contents 39% (P < 0.05). Endothelial (eNOS) was detected in luminal endothelium, whereas neuronal NOS (nNOS) protein was only in the media. A second group of ewes received E(2)beta (1 microg/kg iv) for 4 days and acute intravenous E(2)beta (n = 8) or vehicle (n = 4) on day 5. UBF rose 5.5-fold (P < 0.001) 115 min after E(2)beta, at which time only endothelium derived calcium-dependent NOS activity increased 30 +/- 13% (P < 0.05). Daily E(2)beta enhances basal and E(2)beta-mediated increases in UBF, which parallel increases in endothelium-derived eNOS and smooth muscle-derived nNOS. Acute E(2)beta, however, selectively increases endothelium-derived eNOS. PMID- 10843914 TI - Phosphorylation at tyrosine-524 influences nuclear accumulation of HSP72 with heat stress. AB - Nuclear accumulation of heat shock protein (HSP) 72 occurs after cardiac ischemia. This nuclear accumulation of HSP72 with stress occurs in other tissues and species. We postulated that nuclear accumulation of HSP72 was important for the protective effect of HSP72 and that phosphorylation of a single tyrosine (Y(524)) regulated nuclear accumulation of HSP72. Western blots of immunoprecipitated HSP72 from Cos-1 cells demonstrated that tyrosine becomes phosphorylated after heat shock. Treatment with the tyrosine kinase inhibitor geldanamycin blocked nuclear accumulation of HSP72 with heat shock. Two epitope tagged constructs were made: M17 converting Y(524) to aspartic acid (pseudophosphorylation) and M18 converting Y(524) to phenylalanine. When transfected into Cos-1 cells, M17 accumulates more rapidly and M18 less rapidly than wild-type (WT) HSP72 in the nucleus following heat shock. Cells expressing M18 had less viability after heat shock at 43.5 degrees C than other constructs. After heat shock at 45 degrees C, cells expressing M17 had superior survival compared with WT and M18. These data suggest that phosphorylation at Y(524) facilitates nuclear accumulation of HSP72 following heat stress, and substitution of aspartic acid at Y(524) enhances resistance to heat-shock injury. PMID- 10843916 TI - A self-powered constant infusion device for use in unrestrained rats. AB - We developed a device that delivers fluid through a catheter at a constant rate and can be used in conscious animals to solve a variety of problems. For example, this device can be used for delivering drugs and maintaining intravascular catheter patency. The device provides infusions at low flows (1.0-1.5 ml/day), so that experimental agents may be administered with minimal volume loading of the rat. Arterial and venous catheter patency is maintained by infusion of heparinized saline through indwelling catheters attached to the device. The catheters exit from the rat in the intrascapular area and are routed through a protective spring to the device, which is suspended above the cage. The catheters may be attached to pressure transducers, blood may be sampled, and injections or infusions may be made without disturbing the rat. Because the device is self contained, it can be suspended by a fluid-free swivel that rotates through 360 degrees, providing minimal restraint. The device has been used successfully to measure arterial and central venous blood pressures in two studies using rats. PMID- 10843915 TI - A custom confocal and two-photon digital laser scanning microscope. AB - We describe a custom one-photon (confocal) and two-photon all-digital (photon counting) laser scanning microscope. The confocal component uses two avalanche photodiodes (APDs) as the fluorescence detector to achieve high sensitivity and to overcome the limited photon counting rate of a single APD ( approximately 5 MHz). The confocal component is approximately nine times more efficient than our commercial confocal microscope (fluorophore fluo 4). Switching from one-photon to two-photon excitation mode (Ti:sapphire laser) is accomplished by moving a single mirror beneath the objective lens. The pulse from the Ti:sapphire laser is 109 fs in duration at the specimen plane, and average power is approximately 5 mW. Two photon excited fluorescence is detected by a fast photomultiplier tube. With a x63 1.4 NA oil-immersion objective, the resolution of the confocal system is 0.25 microm laterally and 0.52 microm axially. For the two-photon system, the corresponding values are 0.28 and 0.82 microm. The system is advantageous when excitation intensity must be limited, when fluorescence is low, or when thick, scattering specimens are being studied (with two-photon excitation). PMID- 10843917 TI - Entropies of short binary sequences in heart period dynamics. AB - Dynamic aspects of R-R intervals have often been analyzed by means of linear and nonlinear measures. The goal of this study was to analyze binary sequences, in which only the dynamic information is retained, by means of two different aspects of regularity. R-R interval sequences derived from 24-h electrocardiogram (ECG) recordings of 118 healthy subjects were converted to symbolic binary sequences that coded the beat-to-beat increase or decrease in the R-R interval. Shannon entropy was used to quantify the occurrence of short binary patterns (length N = 5) in binary sequences derived from 10-min intervals. The regularity of the short binary patterns was analyzed on the basis of approximate entropy (ApEn). ApEn had a linear dependence on mean R-R interval length, with increasing irregularity occurring at longer R-R interval length. Shannon entropy of the same sequences showed that the increase in irregularity is accompanied by a decrease in occurrence of some patterns. Taken together, these data indicate that irregular binary patterns are more probable when the mean R-R interval increases. The use of surrogate data confirmed a nonlinear component in the binary sequence. Analysis of two consecutive 24-h ECG recordings for each subject demonstrated good intraindividual reproducibility of the results. In conclusion, quantification of binary sequences derived from ECG recordings reveals properties that cannot be found using the full information of R-R interval sequences. PMID- 10843918 TI - Expiratory flow limitation: Roger S. Mitchell Lecture. PMID- 10843919 TI - Introduction: mechanisms of COPD. PMID- 10843920 TI - Animal models for COPD. PMID- 10843921 TI - A role for collagenase (Matrix metalloproteinase-1) in pulmonary emphysema PMID- 10843922 TI - Experimental evidence from an animal model of adenocarcinoma that chronic inflammation enhances lung cancer risk PMID- 10843923 TI - Lung elastin and matrix. PMID- 10843924 TI - Activation of an embryonic gene product in pulmonary emphysema : identification of the secreted frizzled-related protein PMID- 10843925 TI - Effect of initial collagen concentration on fibroblast mediated contraction of collagen gels PMID- 10843926 TI - Effects of all-trans-retinoic acid in promoting alveolar repair. PMID- 10843927 TI - Cigarette smoke extract decreases the expression of vascular endothelial growth factor by cultured cells and triggers apoptosis of pulmonary endothelial cells PMID- 10843928 TI - Can retinoic acid ameliorate the physiologic and morphologic effects of elastase instillation in the rat? PMID- 10843929 TI - Clinical relevance summary: Collagen vs elastin in pathogenesis of emphysema; cellular origin of elastases; bronchiolitis vs emphysema as a cause of airflow obstruction. PMID- 10843930 TI - A mouse model of cigarette smoke-induced emphysema PMID- 10843931 TI - Pulmonary mechanics of papain emphysema in dogs PMID- 10843933 TI - A study on the effect of cadmium on human lung fibroblasts PMID- 10843932 TI - A cluster analysis of CT scans predicts patients who respond to lung volume reduction surgery PMID- 10843934 TI - Glutathione can protect A549 from smoke-induced inhibition of collagen gel contraction PMID- 10843935 TI - Spontaneous emphysema in surfactant protein D gene-targeted mice PMID- 10843936 TI - Polymorphisms of surfactant protein gene A, B, D, and of SP-B-linked microsatellite markers in COPD of a Mexican population. PMID- 10843937 TI - Cognitive improvement following rehabilitation in patients with COPD PMID- 10843938 TI - Lung neutrophil burden correlates with increased pro-inflammatory cytokines and decreased lung function in individuals with alpha(1)-antitrypsin deficiency PMID- 10843939 TI - Comparison of the structural and inflammatory features of COPD and asthma. Giles F. Filley Lecture. AB - At least three conditions contribute to COPD. (1) Chronic bronchitis (mucous hypersecretion) is an inflammatory condition in which CD8+ T-lymphocytes, neutrophils, and CD68+ monocytes/macrophages predominate. The condition is defined clinically by the presence of chronic cough and recurrent increases in bronchial secretions sufficient to cause expectoration. There is enlargement of mucus-secreting glands and goblet cell hyperplasia, which can occur in the absence of airflow limitation. (2) Adult chronic bronchiolitis (small or peripheral airways disease) is an inflammatory condition of small bronchi and bronchioli in which there are predominantly CD8+ and pigmented macrophages. The functional defect is difficult to detect clinically but may be recognized by sophisticated tests of small airway function. There is mucous metaplasia, enlargement of the mass of bronchiolar smooth muscle, and loss of alveolar attachments. (3) Emphysema is an inflammatory condition of the alveoli in which T lymphocytes, neutrophils, and pigmented alveolar macrophages are involved, associated with the release of excessive amounts of elastases. It is defined anatomically by permanent, destructive enlargement of airspaces distal to terminal bronchioli without obvious fibrosis. In contrast, asthma is a clinical syndrome characterized by allergic inflammation of bronchi and bronchioli in which CD4+ (helper) T-lymphocytes and eosinophils predominate. There is increased production and release of interleukin (IL)-4 and IL-5, which is referred to as a Th2-type response. There is usually increased tracheobronchial responsiveness to a variety of stimuli, and the condition is usually manifest as variable airflow obstruction. While differences between COPD and asthma have been highlighted, new data are emerging that indicate there may also be similarities. PMID- 10843940 TI - Morphometry explains variation in airway responsiveness in transgenic mice overexpressing interleukin-6 and interleukin-11 in the lung. PMID- 10843941 TI - Expression of 15-lipoxygenase and evidence for apoptosis in the lungs from patients with COPD PMID- 10843942 TI - Mechanisms of airway hypersecretion and novel therapy. PMID- 10843943 TI - Ongoing airway inflammation in patients with COPD who Do not currently smoke PMID- 10843944 TI - Myristoylated alanine-rich C-kinase substrate protein : A major intracellular regulatory molecule controlling secretion of mucin by human airway goblet cells PMID- 10843945 TI - Skeletal muscle function in COPD. AB - Few effective therapies exist for patients with COPD. Rehabilitative therapy aimed at curing dysfunction of the peripheral muscles may be an appropriate addition to this short list. This review does the following: (1) presents evidence that skeletal muscle dysfunction is present in COPD patients; (2) considers the mechanisms of this dysfunction; (3) describes the role of exercise training in correcting this disorder; and (4) speculates that anabolic hormone supplementation may find a place in COPD therapy. Further research will be necessary to refine these concepts. PMID- 10843946 TI - Response of human airway epithelium In vitro to inflammatory mediators : dependence on the state of cellular differentiation PMID- 10843947 TI - Chronic alveolar hypoventilation helps to maintain the inspiratory muscle effort of COPD patients within sustainable limits. PMID- 10843948 TI - Familial aggregation of severe, early-onset COPD : candidate gene approaches PMID- 10843950 TI - Comparison of bone mineral density in elderly female patients with COPD and bronchial asthma PMID- 10843951 TI - Exhaled nitric oxide and induced sputum analysis as markers of airway inflammation in subjects with COPD PMID- 10843949 TI - Nutrition and metabolism in COPD. PMID- 10843952 TI - Inhibition of vascular endothelial growth factor receptors causes emphysema in rats PMID- 10843953 TI - Latent adenoviral infection in the pathogenesis of emphysema: the Parker B. Francis Lectureship. PMID- 10843954 TI - Proteases and chemotactic factors in BAL fluid from subjects with subclinical emphysema PMID- 10843955 TI - Airway inflammation and hyperresponsiveness to adenosine 5'-monophosphate in COPD PMID- 10843956 TI - HIV infection increases susceptibility to smoking-induced emphysema PMID- 10843957 TI - Bacterial infection and the pathogenesis of COPD. AB - Bacterial infection of the lower respiratory tract can impact on the etiology, pathogenesis, and the clinical course of COPD in several ways. Several recent cohort studies suggest that lung growth is impaired by childhood lower respiratory tract infection, making these individuals more vulnerable to developing COPD on exposure to additional injurious agents. Impairment of mucociliary clearance and local immune defense in smokers allows bacterial pathogens to gain a foothold in the lower respiratory tract. These pathogens and their products can cause further impairment of mucociliary clearance due to enhanced mucus secretion, disruption of normal ciliary activity, and airway epithelial injury, and thus persist in the lower respiratory tract. This chronic colonization of the lower respiratory tract by bacterial pathogens could induce a chronic inflammatory response with lung damage. Nontypeable Haemophilus influenzae, usually regarded as an extracellular mucosal pathogen, has been demonstrated to cause intracellular infections of the upper and lower respiratory tract respiratory tissue. Increased incidence of chronic Chlamydia pneumoniae infection of the respiratory tract has been associated with COPD. These chronic infections of respiratory tissues could contribute to the pathogenesis of COPD by altering the host response to cigarette smoke or by inducing a chronic inflammatory response. Application of newer molecular and immunologic research techniques is helping us define precisely the role of bacterial infection in COPD. PMID- 10843959 TI - The role of haemophilus parainfluenzae in COPD PMID- 10843958 TI - Bronchial inflammation: its relationship to colonizing microbial load and alpha(1)-antitrypsin deficiency. AB - Neutrophil elastase is capable of generating many of the features of chronic bronchial disease. In patients with COPD, airways inflammation with neutrophil recruitment and elastase release is positively correlated with colonizing bacterial load in the stable clinical state (p < 0.0005). In addition, alpha(1) antitrypsin deficiency is associated with a greater neutrophil load, higher elastase activity, leukotriene-B(4) concentration, and serum protein leak than matched patients without deficiency (p < 0.005). These data confirm an effect of bronchial colonization on airways inflammation in COPD and indicate the role of alpha(1)-antitrypsin in its modulation. PMID- 10843960 TI - Mechanisms of COPD: clinical relevance of research presentations at the Thomas Petty Aspen Lung Conference 1999. PMID- 10843961 TI - Protease injury in the development of COPD: Thomas A. Neff Lecture. PMID- 10843963 TI - The dutch hypothesis PMID- 10843962 TI - The combination of elastase and sulfur dioxide exposure causes COPD-like lesions in the rat. PMID- 10843964 TI - Evidence for neutrophil involvement in the development of subclinical emphysema PMID- 10843966 TI - Quantum proteolysis by neutrophils : implications for pulmonary emphysema in alpha(1)-antitrypsin deficiency PMID- 10843965 TI - Oxidants/antioxidants and COPD. AB - Oxidative stress results from an oxidant/antioxidant imbalance, an excess of oxidants and/or a depletion of antioxidants. Oxidative stress is thought to play an important role in the pathogenesis of a number of lung diseases, not only through direct injurious effects, but by involvement in the molecular mechanisms that control lung inflammation. A number of studies have shown an increased oxidant burden and consequently increased markers of oxidative stress in the airspaces, breath, blood, and urine in smokers and in patients with COPD. The presence of oxidative stress has important consequences for the pathogenesis of COPD. These include oxidative inactivation of antiproteinases, airspace epithelial injury, increased sequestration of neutrophils in the pulmonary microvasculature, and gene expression of proinflammatory mediators. With regard to the latter, oxidative stress has a role in enhancing the inflammation that occurs in smokers and patients with COPD, through the activation of redox sensitive transcriptions factors such as nuclear factor-kappaB and activator protein-1, which regulate the genes for proinflammatory mediators and protective antioxidant gene expression. The sources of the increased oxidative stress in patients with COPD are derived from the increased burden of oxidants present in cigarette smoke, or from the increased amounts of reactive oxygen species released from leukocytes, both in the airspaces and in the blood. Antioxidant depletion or deficiency in antioxidants may contribute to oxidative stress. The development of airflow limitation is related to dietary deficiency of antioxidants, and hence dietary supplementation may be a beneficial therapeutic intervention in this condition. Antioxidants that have good bioavailability or molecules that have antioxidant enzyme activity may be therapies that not only protect against the direct injurious effects of oxidants, but may fundamentally alter the inflammatory events that play an important part in the pathogenesis of COPD. PMID- 10843968 TI - Mechanisms of COPD: conference summary. PMID- 10843967 TI - Neutrophil elastase induces MUC5AC messenger RNA expression by an oxidant dependent mechanism. AB - Airway diseases such as cystic fibrosis, chronic bronchitis, and viral- or pollution-triggered asthma have two common pathologic features: mucus obstruction of the airways, and neutrophil-predominant airway inflammation. Neutrophils release high concentrations of elastase (neutrophil elastase [NE]), a serine protease, into the airways; exposure to elastase results in secretory metaplasia and increased production/secretion of mucin glycoproteins. We have previously shown that NE increases gene expression of a respiratory mucin, MUC5AC, in both A549, a lung adenocarcinoma cell line, and cultured normal human bronchial epithelial cells. In this study, we explored the intracellular signaling mechanisms required for NE-regulated MUC5AC gene expression. A549 cells were treated with dimethylthiourea (DMTU; 4 mM and 40 mM), a scavenger of hydroxyl radical, peroxynitrite, and other hydroxylated products, prior to and during NE stimulation. DMTU inhibited NE-induced MUC5AC expression. Furthermore, using dichlorodihydrofluorescein, an intracellular redox indicator, we showed that in both A549 cells and cultured normal human bronchial epithelial cells, NE treatment induced oxidative stress. These results support the role of reactive oxygen species mediating NE-induced MUC5AC gene expression. PMID- 10843969 TI - Apoptosis of alveolar macrophages by cigarette smoke PMID- 10843970 TI - COPD: Working towards a greater understanding. Introduction. PMID- 10843971 TI - Scope of the COPD problem in North America: early studies of prevalence and NHANES III data: basis for early identification and intervention. AB - COPD is a common disease that is often not diagnosed in the primary-care setting. This review highlights the findings of studies ranging from the early 1960s to the third National Health and Nutrition Examination Survey (NHANES III) study, undertaken to try and accurately estimate the prevalence of COPD in the US population. Results of the NHANES III indicate that COPD remains frequently underdiagnosed, and that spirometry should be more widely used in the primary care setting to identify asymptomatic disease. Early intervention, in particular smoking cessation, could alter the course and outcome of disease in patients with COPD. PMID- 10843972 TI - COPD: the scope of the problem in Europe. AB - In 1995, the European Respiratory Society published a European Consensus Statement on the optimal assessment and management of COPD. In the document, several important areas for future research are identified that may help to increase knowledge of the current situation of COPD within Europe; these include pathophysiology, epidemiology, and the clinical benefits of treatment. This article reviews a selection of important data that have become available since the consensus statement was published, with a specific focus on epidemiology and treatment. PMID- 10843973 TI - International efforts directed at attacking the problem of COPD. AB - COPD is the only leading cause of death that is increasing in prevalence worldwide. The lack of international standardization in the diagnosis of COPD means that intercountry comparisons are difficult. This review highlights the Global Initiative for Obstructive Lung Disease, a program aimed at focusing attention on the importance of COPD as a global health problem, and designing and implementing consistent international strategies for effective prevention, diagnosis, and treatment. PMID- 10843974 TI - Prevalence of airways obstruction in a general population: European Respiratory Society vs American Thoracic Society definition. AB - STUDY OBJECTIVES: To evaluate the distribution of airways obstruction in a general population sample. METHODS: Cross-sectional epidemiologic survey of a general population sample living in Po Delta area (North Italy). Data on respiratory symptoms, diseases, and risk factors were collected through standardized interviewer-administered questionnaires. Lung function tests were performed, with criteria for defining airways obstruction based on the 1995 European Respiratory Society (ERS) statement (FEV(1)/vital capacity ratio < 88% predicted and < 89% predicted in men and women, respectively), "clinical" criteria (FEV(1)/FVC ratio < 70%), and the 1986 American Thoracic Society (ATS) statement (FEV(1)/FVC ratio < 75%). RESULTS: A total of 1,727 subjects aged > 25 years investigated from 1988 to 1991 were included. Prevalence rates of airways obstruction for subjects 25 to 45 years old and subjects >/= 46 years old were as follows: ERS, 10.8% and 12.2%; clinical, 9.9% and 28.8%; and ATS, 27% and 57%, respectively. When considering only moderate/severe obstruction, the rates were as follows: ERS, 0.4% and 3.6%; clinical, 0.3% and 4.4%; and ATS, 0.5% and 5.2%, respectively. The trend was confirmed after stratifying for smoking habit and the presence/absence of respiratory symptoms/diseases. The highest specificity and predictive value for any respiratory symptom/disease was shown by the ERS, and the lowest was shown by the ATS criterion, while the reverse was true for sensitivity; overall accuracy was slightly lower for the ATS criterion. Multiple logistic regression models indicated a higher number of significant associations with known risk factors for airways obstruction according to clinical and ATS criteria than ERS criterion. CONCLUSIONS: The prevalence of COPD in a general population depends very much on the criterion used for definition of airways obstruction. Further research is needed to reach a standardized and epidemiologically consistent criterion for airways obstruction. PMID- 10843975 TI - Utilization in COPD: patient characteristics and diagnostic evaluation. AB - STUDY OBJECTIVES: Information on current practices of COPD diagnosis and treatment is needed to identify opportunities for improving care. This study describes the clinical characteristics and diagnostic evaluations of COPD patients in a health maintenance organization (HMO) and a university-affiliated county medical center (UMC). DESIGN: Cross-sectional survey performed in a 174,484-member regional HMO and in The University of New Mexico Hospitals and Clinics (UNMH). PATIENTS: Two hundred COPD patients from each system randomly selected from administrative databases based on discharge diagnoses. RESULTS: COPD patients in the UMC, compared to those in the HMO, were younger (mean age, 59.3 vs 66.9 years, respectively), were more likely to be using home oxygen (33% vs 20%, respectively), and had fewer chronic medical conditions (mean number of conditions, 3.1 vs 3.7, respectively) (p < 0.01 for all differences). Approximately half of the COPD patients in both groups continued to smoke cigarettes during the study year. Only 38% of patients in the HMO and 42% in the UNMH system had spirometry results documented in their medical records. CONCLUSIONS: The demographic and clinical characteristics of the COPD patients in these two health-care systems were very different, but smoking status and utilization of diagnostic tests were similar. The diagnosis of COPD in most patients was based only on a history of chronic respiratory symptoms and smoking; spirometry often was not used to confirm the diagnosis. An increased emphasis on smoking cessation and more effective utilization of spirometry are needed to improve the management of COPD in these health-care systems. PMID- 10843976 TI - The prevalence of COPD: using smoking rates to estimate disease frequency in the general population. AB - STUDY OBJECTIVES: To develop and validate a model based on smoking rates that will provide reliable estimates of the true prevalence of COPD that include both clinically detected and undetected patients. DESIGN: Model based on literature review. Age- and gender-specific rates of lung impairment by smoking status were applied to US smoking data. Resultant estimates were compared to the actual prevalence of obstructive airway disease as estimated by US national surveys. The model then was applied to estimate the prevalence of COPD in several European countries, where national data on undiagnosed lung disease do not exist. SETTING: The model was adapted from both a literature review and health-care data, and the analysis was applied to the United States and Europe. RESULTS: Using smoking rates, we estimate from our model that 15.3 million people who are > 40 years of age in the United States have COPD. The prevalence estimate, based on spirometric definitions for COPD in the same age group using the Third National Health and Nutrition Examination Survey (NHANES III), is 17.1 million people. NHANES III and other US national health-care surveys further suggest that only between 2.4 and 7 million people actually have COPD diagnosed; thus, the proportion of COPD that is currently being diagnosed in the United States is between 14% and 46% of all cases. Using smoking rates and our model, which was developed and validated for the United States, we calculated the prevalence of COPD for Germany (2.7 million people), the United Kingdom (3.0 million people), Spain (1.5 million people), Italy (2.6 million people), and France (2.6 million people) in those people > 45 years of age. CONCLUSIONS: Smoking rates appear to provide a useful method of estimating current COPD prevalence in those countries where more objective data are unavailable. These results are important because recognition of the true burden of disease and corresponding efforts to increase early identification of COPD can help to reduce the morbidity and mortality associated with COPD in populations at risk. PMID- 10843977 TI - Smoking cessation. AB - Cessation of cigarette smoking is the single most important therapeutic intervention that is effective in reducing the symptoms of COPD and in preventing its onset. Smoking cessation is, therefore, a major goal in efforts to mitigate the burden of this disease. This review will consider the pharmacologic and behavioral therapies that have been used to assist smokers in overcoming their addiction. These strategies assist a significant minority of smokers to stop smoking and, thus, they can have an important positive impact on COPD as well as on other health outcomes. PMID- 10843978 TI - COPD: early detection and intervention. AB - The compelling evidence for the increasing economic and social burden of COPD, resulting from its rising prevalence and significant morbidity, has been reviewed in other sections of this supplement. The impact of this disease within the United States and globally is projected to increase irrespective of short-term medical action, but developing successful strategies to identify the illness and reduce its impact is essential if this growing problem is to be managed successfully. In this article, some of the important concepts relevant to this process are considered, and some of the present techniques used to intervene in established COPD are reviewed. PMID- 10843979 TI - Raising awareness of COPD in primary care. AB - COPD is a major cause of mortality and a significant drain on health-care resources but is widely underdiagnosed in the primary-care setting. There is an urgent need to raise the profile of the disease among both primary-care physicians and patients. At the workshop "COPD: Working Towards a Greater Understanding," a panel of COPD experts from Europe and the United States discussed ways in which awareness of COPD could be raised. Access to spirometry, and education in its use and relevance, was identified as a major goal for primary-care physicians. Simple questionnaires can promote patient awareness and provide feedback to physicians. COPD needs to be identified as not just a disease of smokers. PMID- 10843980 TI - COPD: exacerbation. AB - A renewed interest in the clinical and pathogenic aspects of COPD exacerbation is timely in view of national and global COPD initiatives. The three big problems regarding COPD continue to be the following: prevention of the disease; slowing progression of the disease once diagnosis has been established; and prevention and more effective treatment of the so-called exacerbation. The following assessment will raise more questions than answers and will review some of the past and current concepts and contexts. PMID- 10843981 TI - Infectious etiology of acute exacerbations of chronic bronchitis. AB - Infectious agents are a major cause of acute exacerbations of chronic bronchitis (AECB) and COPD. Several respiratory viruses are associated with 30% of exacerbations, with or without a superimposed bacterial infection. Atypical bacteria, mostly Chlamydia pneumoniae, have been implicated in < 10% of AECB. The role of bacterial pathogens when isolated from the respiratory tract during AECB has become better defined by application of several newer investigative techniques. Bacterial pathogens can be isolated in significant concentrations from distal airways in 50% of AECB. Specific immune responses to surface exposed antigens of the infecting pathogen have been shown to develop after an exacerbation. Emerging evidence from molecular epidemiology and measurement of airway inflammation further support the role of bacteria in AECB. When properly defined, 80% of AECB are likely to be infectious in origin. PMID- 10843982 TI - Role of neutrophil elastase in hypersecretion during COPD exacerbations, and proposed therapies. AB - A common feature of COPD and other chronic lung diseases is hypersecretion of mucus into the airways, causing peripheral airway plugging and further airflow obstruction. The mucus is secreted by goblet cells, which are present in excessive numbers in COPD. This review describes how neutrophils in the airways of COPD patients stimulate the goblet cells to secrete their products. Recent findings on the mechanisms of neutrophil stimulation of goblet cell degranulation are discussed. These implicate the proteolytic enzyme elastase and cell surface adhesion molecules, and provide a basis for the investigation of potential novel therapies. PMID- 10843983 TI - Exacerbations of COPD: environmental mechanisms. AB - Air pollution as a trigger for exacerbations of COPD has been recognized for > 50 years, and has led to the development of air quality standards in many countries that substantially decreased the levels of air pollutants derived from the burning of fossil fuels, such as black smoke and sulfur dioxide. However, the recent dramatic increase in motor vehicle traffic has produced a relative increase in the levels of newer pollutants, such as ozone and fine-particulate air pollution < 10 microm in diameter. Numerous epidemiologic studies have shown associations between the levels of these air pollutants and adverse health effects, such as exacerbations of airways diseases and even deaths from respiratory and cardiovascular causes. Elucidation of the mechanism of the harmful effects of these pollutants should allow improved risk assessment for patients with airways diseases who are be susceptible to the effects of these air pollutants. PMID- 10843984 TI - Toward a consensus definition for COPD exacerbations. AB - In patients with COPD, an acute worsening of respiratory symptoms is often described as an exacerbation. Exacerbations are associated with a significant increase in mortality, hospitalization, and health-care utilization, but there is currently no widely accepted definition of what constitutes an exacerbation of COPD. This paper summarizes the discussions of the workshop, "COPD: Working Towards a Greater Understanding," in which the participants proposed the following working definition of an exacerbation of COPD: a sustained worsening of the patient's condition, from the stable state and beyond normal day-to-day variations, that is acute in onset and necessitates a change in regular medication in a patient with underlying COPD. PMID- 10843985 TI - The breast cancer susceptibility gene BRCA1 is required for subnuclear assembly of Rad51 and survival following treatment with the DNA cross-linking agent cisplatin. AB - Mutations in breast cancer tumor susceptibility genes, BRCA1 and BRCA2, predispose women to early onset breast cancer and other malignancies. The Brca genes are involved in multiple cellular processes in response to DNA damage including checkpoint activation, gene transcription, and DNA repair. Biochemical interaction with the recombinational repair protein Rad51 (Scully, R., Chen, J., Ochs, R. L., Keegan, K., Hoekstra, M., Feunteun, J., and Livingston, D. M. (1997) Cell 90, 425-435), as well as genetic evidence (Moynahan, M. E., Chiu, J. W., Koller, B. H., and Jasin, M. (1999) Mol. Cell 4, 511-518 and Snouwaert, J. N., Gowen, L. C., Latour, A. M., Mohn, A. R., Xiao, A., DiBiase, L., and Koller, B. H. (1999) Oncogene 18, 7900-7907), demonstrates that Brca1 is involved in recombinational repair of DNA double strand breaks. Using isogenic Brca1(+/+) and brca1(-/-) mouse embryonic stem (ES) cell lines, we investigated the role of Brca1 in the cellular response to two different categories of DNA damage: x-ray induced damage and cross-linking damage caused by the chemotherapeutic agent, cisplatinum. Immunoflourescence studies with normal and brca1(-/-) mutant mouse ES cell lines indicate that Brca1 promotes assembly of subnuclear Rad51 foci following both types of DNA damage. These foci are likely to be oligomeric complexes of Rad51 engaged in repair of DNA lesions or in processes that allow cells to tolerate such lesions during DNA replication. Clonogenic assays show that brca1(-/-) mutants are 5-fold more sensitive to cisplatinum compared with wild-type cells. Our studies suggest that Brca1 contributes to damage repair and/or tolerance by promoting assembly of Rad51. This function appears to be shared with Brca2. PMID- 10843986 TI - Requirement of Ras/MAPK pathway activation by transforming growth factor beta for transforming growth factor beta 1 production in a Smad-dependent pathway. AB - Our previous results have shown that transforming growth factor beta (TGFbeta) rapidly activates Ras, as well as both ERKs and SAPKs. In order to address the biological significance of the activation of these pathways by TGFbeta, here we examined the role of the Ras/MAPK pathways and the Smads in TGFbeta(3) induction of TGFbeta(1) expression in untransformed lung and intestinal epithelial cells. Expression of either a dominant-negative mutant of Ras (RasN17) or a dominant negative mutant of MKK4 (DN MKK4), or addition of the MEK1 inhibitor PD98059, inhibited the ability of TGFbeta(3) to induce AP-1 complex formation at the TGFbeta(1) promoter, and the subsequent induction of TGFbeta(1) mRNA. The primary components present in this TGFbeta(3)-inducible AP-1 complex at the TGFbeta(1) promoter were JunD and Fra-2, although c-Jun and FosB were also involved. Furthermore, deletion of the AP-1 site in the TGFbeta(1) promoter or addition of PD98059 inhibited the ability of TGFbeta(3) to stimulate TGFbeta(1) promoter activity. Collectively, our data demonstrate that TGFbeta(3) induction of TGFbeta(1) is mediated through a signaling cascade consisting of Ras, the MAPKKs MKK4 and MEK1, the MAPKs SAPKs and ERKs, and the specific AP-1 proteins Fra-2 and JunD. Although Smad3 and Smad4 were not detectable in TGFbeta(3)-inducible AP-1 complexes at the TGFbeta(1) promoter, stable expression of dominant-negative Smad3 could significantly inhibit the ability of TGFbeta(3) to stimulate TGFbeta(1) promoter activity. Transient expression of dominant-negative Smad4 also inhibited the ability of TGFbeta(3) to transactivate the TGFbeta(1) promoter. Thus, although the Ras/MAPK pathways are essential for TGFbeta(3) induction of TGFbeta(1), Smads may only contribute to this biological response in an indirect manner. PMID- 10843987 TI - Trypanosoma cruzi surface mucins with exposed variant epitopes. AB - The protozoan parasite Trypanosoma cruzi, the agent of Chagas disease, has a large number of mucin molecules on its surface, whose expression is regulated during the life cycle. These mucins are the main acceptors of sialic acid, a monosaccharide that is required by the parasite to infect and survive in the mammalian host. A large mucin-like gene family named TcMUC containing about 500 members has been identified previously in T. cruzi. TcMUC can be divided into two subfamilies according to the presence or absence of tandem repeats in the central region of the genes. In this work, T. cruzi parasites were transfected with one tagged member of each subfamily. Only the product from the gene with repeats was highly O-glycosylated in vivo. The O-linked oligosaccharides consisted mainly of beta-d-Galp(1-->4)GlcNAc and beta-d-Galp(1-->4)[beta-d-Galp(1-->6)]-d-GlcNAc. The same glycosyl moieties were found in endogenous mucins. The mature product was anchored by glycosylphosphatidylinositol to the plasma membrane and exposed to the medium. Sera from infected mice recognized the recombinant product of one repeats-containing gene thus showing that they are expressed during the infection. TcMUC genes encode a hypervariable region at the N terminus. We now show that the hypervariable region is indeed present in the exposed mature N termini of the mucins because sera from infected hosts recognized peptides having sequences from this region. The results are discussed in comparison with the mucins from the insect stages of the parasite (Di Noia, J. M., D'Orso, I., Sanchez, D. O., and Frasch, A. C. C. (2000) J. Biol. Chem. 275, 10218-10227) which do not have variable regions. PMID- 10843988 TI - High molecular weight kininogen utilizes heparan sulfate proteoglycans for accumulation on endothelial cells. AB - Kininogens, the high molecular weight precursor of vasoactive kinins, bind to a wide variety of cells in a specific, reversible, and saturable manner. The cell docking sites have been mapped to domains D3 and D5(H) of kininogens; however, the corresponding cellular acceptor sites are not fully established. To characterize the major cell binding sites for kininogens exposed by the endothelial cell line EA.hy926, we digested intact cells with trypsin and other proteases and found a time- and concentration-dependent loss of (125)I-labeled high molecular weight kininogen (H-kininogen) binding capacity (up to 82%), indicating that proteins are crucially involved in kininogen cell attachment. Cell surface digestion with heparinases similarly reduced kininogen binding capacity (up to 78%), and the combined action of heparinases and trypsin almost eliminated kininogen binding (up to 85%), suggesting that proteoglycans of the heparan sulfate type are intimately involved. Consistently, inhibitors such as p nitrophenyl-beta-d-xylopyranoside and chlorate interfering with heparan sulfate proteoglycan biosynthesis reduced the total number of kininogen binding sites in a time- and concentration-dependent manner (up to 67%). In vitro binding studies demonstrated that biotinylated H-kininogen binds to heparan sulfate glycosaminoglycans via domains D3 and D5(H) and that the presence of Zn(2+) promotes this association. Cloning and over-expression of the major endothelial heparan sulfate-type proteoglycans syndecan-1, syndecan-2, syndecan-4, and glypican in HEK293t cells significantly increased total heparan sulfate at the cell surface and thus the number of kininogen binding sites (up to 3. 3-fold). This gain in kininogen binding capacity was completely abolished by treating transfected cells with heparinases. We conclude that heparan sulfate proteoglycans on the surface of endothelial cells provide a platform for the local accumulation of kininogens on the vascular lining. This accumulation may allow the circumscribed release of short-lived kinins from their precursor molecules in close proximity to their sites of action. PMID- 10843989 TI - The role of Mg2+ cofactor in the guanine nucleotide exchange and GTP hydrolysis reactions of Rho family GTP-binding proteins. AB - The biological activities of Rho family GTPases are controlled by their guanine nucleotide binding states in cells. Here we have investigated the role of Mg(2+) cofactor in the guanine nucleotide binding and hydrolysis processes of the Rho family members, Cdc42, Rac1, and RhoA. Differing from Ras and Rab proteins, which require Mg(2+) for GDP and GTP binding, the Rho GTPases bind the nucleotides in the presence or absence of Mg(2+) similarly, with dissociation constants in the submicromolar concentration. The presence of Mg(2+), however, resulted in a marked decrease in the intrinsic dissociation rates of the nucleotides. The catalytic activity of the guanine nucleotide exchange factors (GEFs) appeared to be negatively regulated by free Mg(2+), and GEF binding to Rho GTPase resulted in a 10-fold decrease in affinity for Mg(2+), suggesting that one role of GEF is to displace bound Mg(2+) from the Rho proteins. The GDP dissociation rates of the GTPases could be further stimulated by GEF upon removal of bound Mg(2+), indicating that the GEF-catalyzed nucleotide exchange involves a Mg(2+) independent as well as a Mg(2+)-dependent mechanism. Although Mg(2+) is not absolutely required for GTP hydrolysis by the Rho GTPases, the divalent ion apparently participates in the GTPase reaction, since the intrinsic GTP hydrolysis rates were enhanced 4-10-fold upon binding to Mg(2+), and k(cat) values of the Rho GTPase-activating protein (RhoGAP)-catalyzed reactions were significantly increased when Mg(2+) was present. Furthermore, the p50RhoGAP specificity for Cdc42 was lost in the absence of Mg(2+) cofactor. These studies directly demonstrate a role of Mg(2+) in regulating the kinetics of nucleotide binding and hydrolysis and in the GEF- and GAP-catalyzed reactions of Rho family GTPases. The results suggest that GEF facilitates nucleotide exchange by destabilizing both bound nucleotide and Mg(2+), whereas RhoGAP utilizes the Mg(2+) cofactor to achieve high catalytic efficiency and specificity. PMID- 10843990 TI - Regulation of the Ca2+-inhibitable adenylyl cyclase type VI by capacitative Ca2+ entry requires localization in cholesterol-rich domains. AB - The endogenous Ca(2+)-inhibitable adenylyl cyclase type VI of C6-2B glioma cells is regulated only by capacitative Ca(2+) entry and not by a substantial elevation of [Ca(2+)](i) from either intracellular stores or via ionophore-mediated Ca(2+) entry (Chiono, M., Mahey, R., Tate, G., and Cooper, D. M. F. (1995) J. Biol. Chem. 270, 1149-1155; Fagan, K. A., Mons, N., and Cooper, D. M. F. (1998) J. Biol. Chem. 273, 9297-9305). The present studies explored the role of cholesterol rich domains in maintaining this functional association. The cholesterol-binding agent, filipin, profoundly inhibited adenylyl cyclase activity. Depletion of plasma membrane cholesterol with methyl-beta-cyclodextrin did not affect forskolin-stimulated adenylyl cyclase activity and did not affect capacitative Ca(2+) entry. However, cholesterol depletion completely ablated the regulation of adenylyl cyclase by capacitative Ca(2+) entry. Repletion of cholesterol restored the sensitivity of adenylyl cyclase to capacitative Ca(2+) entry. Adenylyl cyclase catalytic activity and immunoreactivity were extracted into buoyant caveolar fractions with Triton X-100. The presence of adenylyl cyclase in such structures was eliminated by depletion of plasma membrane cholesterol. Altogether, these data lead us to conclude that adenylyl cyclase must occur in cholesterol-rich domains to be susceptible to regulation by capacitative Ca(2+) entry. These findings are the first indication of regulatory significance for the localization of adenylyl cyclase in caveolae. PMID- 10843991 TI - The protein-tyrosine phosphatase SHP-2 is required during angiotensin II-mediated activation of cyclin D1 promoter in CHO-AT1A cells. AB - Angiotensin II (Ang II) binds to specific G protein-coupled receptors and is mitogenic in Chinese hamster ovary (CHO) cells stably expressing a rat vascular angiotensin II type 1A receptor (CHO-AT(1A)). Cyclin D1 protein expression is regulated by mitogens, and its assembly with the cyclin-dependent kinases induces phosphorylation of the retinoblastoma protein pRb, a critical step in G(1) to S phase cell cycle progression contributing to the proliferative responses. In the present study, we found that in CHO-AT(1A) cells, Ang II induced a rapid and reversible tyrosine phosphorylation of various intracellular proteins including the protein-tyrosine phosphatase SHP-2. Ang II also induced cyclin D1 protein expression in a phosphatidylinositol 3-kinase and mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK)-dependent manner. Using a pharmacological and a co-transfection approach, we found that p21(ras), Raf-1, phosphatidylinositol 3-kinase and also the catalytic activity of SHP-2 and its Src homology 2 domains are required for cyclin D1 promoter/reporter gene activation by Ang II through the regulation of MAPK/ERK activity. Our findings suggest for the first time that SHP-2 could play an important role in the regulation of a gene involved in the control of cell cycle progression resulting from stimulation of a G protein-coupled receptor independently of epidermal growth factor receptor transactivation. PMID- 10843992 TI - N-Formyl peptide receptor ligation induces rac-dependent actin reorganization through Gbeta gamma subunits and class Ia phosphoinositide 3-kinases. AB - The N-formyl peptide receptor is a G protein-coupled transmembrane receptor involved in stimulating a variety of differential responses in neutrophils including chemotaxis, degranulation, superoxide production, transcriptional activation, and actin reorganization. Although it is known that N-formyl-Met-Leu Phe induces actin reorganization, the sequence of events from the receptor to the actin cytoskeleton is not well characterized. To study the signaling pathway from the N-formyl peptide receptor to the actin cytoskeleton, we developed a model system utilizing microinjection techniques with a nonhematopoietic cell line. An expression vector coding for the N-formyl peptide receptor was microinjected into porcine aortic endothelial cells and stimulated with N-formyl-Met-Leu-Phe to induce actin reorganization and membrane ruffling. The receptor-mediated signal was blocked by pertussis toxin and by a dominant negative Rac-N17, indicating the involvement of G(i)alpha subunit and the small guanosine triphosphatase Rac, respectively. Moreover, Gbetagamma subunits and membrane targeted forms of phosphatidylinositol (PI) 3-kinase alpha were sufficient to induce similar actin reorganization, and coexpression of various mutants of PI 3-kinase with the N formyl peptide receptor identified a link to class Ia PI-3 kinase-mediated actin reorganization. PMID- 10843993 TI - Specific glutamine and asparagine residues of gamma-S crystallin are resistant to in vivo deamidation. AB - It has been hypothesized that resistance to nonenzymatic deamidation of asparagine and glutamine residues may be an important determinant of protein stability in vivo. As a test of this hypothesis, we analyzed the central region of old human lenses, which contain proteins such as gamma-S crystallin that were synthesized during the fetal-embryonic periods of development. Total protein from the fetal-embryonic region of old human lenses was digested with trypsin, followed by resolution of tryptic fragments containing amidated and deamidated forms using high pressure liquid chromatography-reverse phase chromatography together with synthetic peptide standards and mass spectral analysis. The results demonstrate no detectable deamidation of glutamine 92, glutamine 96, asparagine 143, and glutamine 170 from gamma-S crystallin from old human lenses, consistent with the hypothesis that very long-lived proteins can contain asparagine and glutamine residues that are extremely resistant to in vivo deamidation. PMID- 10843994 TI - Efficient TGF-beta induction of the Smad7 gene requires cooperation between AP-1, Sp1, and Smad proteins on the mouse Smad7 promoter. AB - Sma- and Mad-related protein 7 (Smad7) is an antagonist of transforming growth factor-beta (TGF-beta) signaling, which has been shown to be induced by TGF-beta itself and also by other stimuli. In an effort to understand the molecular mechanisms underlying the transcriptional regulation of the Smad7 gene by TGF beta, we cloned and functionally characterized a mouse genomic DNA fragment encompassing the mouse Smad7 proximal promoter. This region was found to contain a CpG island and to be devoid of a classical TATA box. Cloned upstream of a promoter-lacking luciferase reporter gene, this region conferred robust TGF-beta induced transcription. Point mutations in a palindromic Smad binding element, abolished TGF-beta inducibility completely. Through the use of electrophoretic mobility shift assays, we showed the presence of Smad2, Smad3, and Smad4 in complexes binding to the Smad binding element. Interestingly, we also found that point mutation and/or deletion of binding sites for the transcription factors activator protein-1 and Sp1 led to an attenuation of the basal promoter activity, as well as of the TGF-beta-mediated induction of Smad7. Taken together, our data imply that Smads, together with activator protein-1 and Sp1 transcription factors, are essential for efficient Smad7 promoter activity. PMID- 10843995 TI - Interleukin-6 family of cytokines mediate angiotensin II-induced cardiac hypertrophy in rodent cardiomyocytes. AB - This study was designed to investigate whether angiotensin II induces the interleukin (IL)-6 family of cytokines in cardiac fibroblasts and, if so, whether these cytokines can augment cardiac hypertrophy. Angiotensin II increased IL-6, leukemia inhibitory factor (LIF) and cardiotrophin-1 mRNA by 6.5-, 10.2-, and 2.0 fold, respectively, but did not affect IL-11, ciliary neurotrophic factor, or oncostatin M in cardiac fibroblasts. Enzyme-linked immunosorbent assay revealed that angiotensin II-stimulated conditioned medium from cardiac fibroblasts contained 9.3 ng/ml IL-6 at 24 h, which was 24-fold higher than the control. It phosphorylated gp130 and STAT3 in cardiomyocytes, which was reduced with RX435 (anti-gp130 blocking antibody). It increased [(3)H]phenylalanine uptake and cell area by 44% and 86% in cardiomyocytes compared with mock medium. RX435 suppressed these increases by 26% and 38%, while TAK044 (endothelin-A/B-R blocker) suppressed them by 52% and 52%, respectively. Antisense oligonucleotides against LIF and cardiotrophin-1 blocked their up-regulation, and attenuated the conditioned medium-induced increase in [(3)H]phenylalanine uptake by 21% and 13%, respectively. The combination of antisense oligonucleotides to LIF and cardiotrophin-1 decreased their uptake by 33%. These results indicated that angiotensin II induced IL-6, LIF, and cardiotrophin-1 in cardiac fibroblasts, and that these cytokines, particularly LIF and cardiotrophin-1, activated gp130 linked signaling and contributed to angiotensin II-induced cardiomyocyte hypertrophy. PMID- 10843996 TI - The zab domain of the human RNA editing enzyme ADAR1 recognizes Z-DNA when surrounded by B-DNA. AB - The Zab domain of the editing enzyme ADAR1 binds tightly and specifically to Z DNA stabilized by bromination or supercoiling. A stoichiometric amount of protein has been shown to convert a substrate of suitable sequence to the Z form, as demonstrated by a characteristic change in the CD spectrum of the DNA. Now we show that Zab can bind not only to isolated Z-forming d(CG)(n) sequences but also to d(CG)(n) embedded in B-DNA. The binding of Zab to such sequences results in a complex including Z-DNA, B-DNA, and two B-Z junctions. In this complex, the d(CG)(n) sequence, but not the flanking region, is in the Z conformation. The presence of Z-DNA was detected by cleavage with a Z-DNA specific nuclease, by undermethylation using Z-DNA sensitive SssI methylase, and by circular dichroism. It is possible that Zab binds to B-DNA with low affinity and flips any favorable sequence into Z-DNA, resulting in a high affinity complex. Alternatively, Zab may capture Z-DNA that exists transiently in solution. The binding of Zab to potential as well as established Z-DNA segments suggests that the range of biological substrates might be wider than previously thought. PMID- 10843997 TI - ADP-ribosylation factor 1 and its activation of phospholipase D are important for the assembly of very low density lipoproteins. AB - The role of ADP-ribosylation factor 1 (ARF-1) in the assembly of very low density lipoproteins (VLDL) was investigated by expressing dominant-negative mutants in McA-RH7777 cells. Transient expression of ARF-1(T31N), a GDP-restrictive mutant, significantly inhibited apolipoprotein B-100 (apoB-100) VLDL production without influencing the biosynthesis of apoB-100 low density lipoproteins or total apoB production (indicating that it inhibited the second step of VLDL assembly) and without altering total protein production or biosynthesis of transferrin, phosphatidylcholine, or triglycerides. These effects were confirmed in stable inducible transfectants. In contrast, expression of an ARF-1 mutant lacking the N terminal 17 amino acids, which has no myristoylation site and cannot interact with the microsomal membrane, did not affect VLDL assembly. Thus, active ARF-1 is needed for the second step of the process. To further explore these observations, we developed a cell-free system based on the postnuclear supernatant isolated from McA-RH7777 cells. In this system, 10-15% of the apoB-100 pool was converted to VLDL in a time- and temperature-dependent way. The assembly process was highly dependent on a heat-stable factor in the d > 1.21 g/ml infranatant of fetal calf serum; this factor was not present in low density lipoproteins or VLDL. Brefeldin A inhibited VLDL assembly in this system, as did a synthetic peptide (corresponding to N-terminal amino acids 2-17 of ARF-1) that displaces ARF-1 from the membrane. Thus, active ARF-1 is also needed for cell-free assembly of VLDL. Guanosine 5'-3-O-(thio)triphosphate also inhibited VLDL assembly in this system, indicating that the process requires ongoing hydrolysis of GTP. 1-Butanol, which inhibits the formation of phosphatidic acid (PA) and instead gives rise to phosphatidylbutanol, inhibited VLDL assembly, whereas 2-butanol, which does not inhibit PA formation, failed to do so. Thus, phospholipase D (PLD)-catalyzed formation of PA from phosphatidylcholine is essential for VLDL assembly. In support of this conclusion, exogenous PLD prevented brefeldin A from inhibiting the assembly process. Our results indicate that ARF-1 participates in the second step of VLDL assembly through a process that involves activation of PLD and production of PA. PMID- 10843998 TI - Transcription termination by RNA polymerase III in fission yeast. A genetic and biochemically tractable model system. AB - In order for RNA polymerase (pol) III to produce a sufficient quantity of RNAs of appropriate structure, initiation, termination, and reinitiation must be accurate and efficient. Termination-associated factors have been shown to facilitate reinitiation and regulate transcription in some species. Suppressor tRNA genes that differ in the dT(n) termination signal were examined for function in Schizosaccharomyces pombe. We also developed an S. pombe extract that is active for tRNA transcription that is described here for the first time. The ability of this tRNA gene to be transcribed in extracts from different species allowed us to compare termination in three model systems. Although human pol III terminates efficiently at 4 dTs and S. pombe at 5 dTs, Saccharomyces cerevisiae pol III requires 6 dTs to direct comparable but lower termination efficiency and also appears qualitatively distinct. Interestingly, this pattern of sensitivity to a minimal dT(n) termination signal was found to correlate with the sensitivity to alpha-amanitin, as S. pombe was intermediate between human and S. cerevisiae pols III. The results establish that the pols III of S. cerevisiae, S. pombe, and human exhibit distinctive properties and that termination occurs in S. pombe in a manner that is functionally more similar to human than is S. cerevisiae. PMID- 10843999 TI - Molecular characterization of human acetyl-CoA synthetase, an enzyme regulated by sterol regulatory element-binding proteins. AB - Through suppressive subtractive hybridization, we identified a new gene whose transcription is induced by sterol regulatory element-binding proteins (SREBPs). The gene encodes acetyl-CoA synthetase (ACS), the cytosolic enzyme that activates acetate so that it can be used for lipid synthesis or for energy generation. ACS genes were isolated previously from yeast, but not from animal cells. Recombinant human ACS was produced by expressing the cloned cDNA transiently in human cells. After purification by nickel chromatography, the 701-amino acid cytosolic enzyme was shown to function as a monomer. The recombinant enzyme produced acetyl-CoA from acetate in a reaction that required ATP. As expected for a gene controlled by SREBPs, ACS mRNA was induced when cultured cells were deprived of sterols and repressed by sterol addition. The pattern of regulation resembled the regulation of enzymes of fatty acid synthesis. ACS mRNA was also elevated in livers of transgenic mice that express dominant-positive versions of all three isoforms of SREBP. We conclude that ACS mRNA, and hence the ability of cells to activate acetate, is regulated by SREBPs in parallel with fatty acid synthesis in animal cells. PMID- 10844000 TI - Modulating protein folding rates in vivo and in vitro by side-chain interactions between the parallel beta strands of green fluorescent protein. AB - We have identified pairs of residues across the two parallel beta strands of green fluorescent protein that facilitate native strand register of the surface exposed beta barrel. After constructing a suitable host environment around two guest residues, minimizing interactions of the guest residues with surrounding side-chains yet maintaining the wild-type protein structure and the chromophore environment, we introduced a library of cross-strand pairings by cassette mutagenesis. Colonies of Escherichia coli transformed with the library differ in intracellular fluorescence. Most of the fluorescent pairs have predominantly charged and polar guest site residues. The magnitude and the rate of fluorescence acquisition in vivo from transformed E. coli cells varies among the mutants despite comparable levels of protein expression. Spectroscopic measurements of purified mutants show that the native protein structure is maintained. Kinetic studies using purified protein with fully matured chromophores demonstrate that the mutants span a 10-fold range in folding rates with undetectable differences in unfolding rates. Thus, green fluorescent protein provides an ideal system for monitoring determinants of in vivo protein folding. Cross-strand pairings affect both protein stability and folding kinetics by favoring the formation of native strand register preferentially to non-native strand alignments. PMID- 10844001 TI - Factor VIIa/tissue factor-induced signaling via activation of Src-like kinases, phosphatidylinositol 3-kinase, and Rac. AB - Tissue factor (TF), apart from activating the extrinsic pathway of the blood coagulation, is a principal regulator of embryonic angiogenesis and oncogenic neoangiogenesis, but also influences inflammation, leukocyte diapedesis and tumor progression. The intracellular domain of TF lacks homology to other classes of receptors and hence the signaling mechanism is poorly understood. Here we demonstrate that factor VIIa (the natural ligand for TF) induces the activation of the Src family members c-Src, Lyn, and Yes, and subsequently phosphatidylinositol 3-kinase (PI3K), followed by stimulation of c-Akt/protein kinase B as well as the small GTPases Rac and Cdc42. In turn Rac mediates p38 mitogen-activated protein (MAP) kinase activation and cytoskeletal reorganization, whereas factor VIIa-induced p42/p44 MAP kinase stimulation required PI3K enzymatic activity but was not inhibited by dominant negative Rac proteins. We propose that this Src family member/PI3K/Rac-dependent signaling pathway is a major mediator of factor VIIa/TF effects in pathophysiology. PMID- 10844002 TI - Defect in peroxisome proliferator-activated receptor alpha-inducible fatty acid oxidation determines the severity of hepatic steatosis in response to fasting. AB - Fasting causes lipolysis in adipose tissue leading to the release of large quantities of free fatty acids into circulation that reach the liver where they are metabolized to generate ketone bodies to serve as fuels for other tissues. Since fatty acid-metabolizing enzymes in the liver are transcriptionally regulated by peroxisome proliferator-activated receptor alpha (PPARalpha), we investigated the role of PPARalpha in the induction of these enzymes in response to fasting and their relationship to the development of hepatic steatosis in mice deficient in PPARalpha (PPARalpha(-/-)), peroxisomal fatty acyl-CoA oxidase (AOX( /-)), and in both PPARalpha and AOX (double knock-out (DKO)). Fasting for 48-72 h caused profound impairment of fatty acid oxidation in both PPARalpha(-/-) and DKO mice, and DKO mice revealed a greater degree of hepatic steatosis when compared with PPARalpha(-/-) mice. The absence of PPARalpha in both PPARalpha(-/-) and DKO mice impairs the induction of mitochondrial beta-oxidation in liver following fasting which contributes to hypoketonemia and hepatic steatosis. Pronounced steatosis in DKO mouse livers is due to the added deficiency of peroxisomal beta oxidation system in these animals due to the absence of AOX. In mice deficient in AOX alone, the sustained hyperactivation of PPARalpha and up-regulation of mitochondrial beta-oxidation and microsomal omega-oxidation systems as well as the regenerative nature of a majority of hepatocytes containing numerous spontaneously proliferated peroxisomes, which appear refractory to store triglycerides, blunt the steatotic response to fasting. Starvation for 72 h caused a decrease in PPARalpha hepatic mRNA levels in wild type mice, with no perceptible compensatory increases in PPARgamma and PPARdelta mRNA levels. PPARgamma and PPARdelta hepatic mRNA levels were lower in fed PPARalpha(-/-) and DKO mice when compared with wild type mice, and fasting caused a slight increase only in PPARgamma levels and a decrease in PPARdelta levels. Fasting did not change the PPAR isoform levels in AOX(-/-) mouse liver. These observations point to the critical importance of PPARalpha in the transcriptional regulatory responses to fasting and in determining the severity of hepatic steatosis. PMID- 10844003 TI - Cooperative interaction between developmentally regulated troponin T and tropomyosin isoforms in the absence of F-actin. AB - Troponin T (TnT) is the tropomyosin (Tm) binding subunit of the troponin complex that mediates the Ca(2+) regulation of actomyosin interaction in striated muscles. Troponin T isoform diversity is marked by a developmentally regulated acidic to basic switch that may modulate muscle contractility. We previously reported that transgenic expression of fast skeletal muscle TnT altered the cooperativity of cardiac muscle. In the present study, we have demonstrated that the binding of acidic TnT to troponin I is weaker than that of basic TnT. However, affinity chromatography experiments showed that Tm bound to acidic TnT with a greater affinity than to basic TnT, consistent with the significantly higher maximal binding of acidic TnT to Tm in solid phase binding assays. Competition and co-immunoprecipitation experiments demonstrated that the binding of TnT to Tm was cooperative in the absence of F-actin. The cooperativity between TnT molecules for Tm binding can be initiated by the conserved COOH-terminal T2 fragment of TnT. This indicates that the interaction of TnT with Tm induces a conformational change in Tm, promoting interaction of TnT with adjacent Tm dimers. This finding suggests a role for TnT and its acidic and basic isoforms in the cooperative release of the inhibition of striated muscle actomyosin interaction. PMID- 10844004 TI - Syntaxin modulation of slow inactivation of N-type calcium channels. AB - Syntaxin, a membrane protein vital in triggering vesicle fusion, interacts with voltage-gated N- and P/Q-type Ca(2+) channels. This biochemical association is proposed to colocalize Ca(2+) channels and presynaptic release sites, thus supporting rapid and efficient initiation of neurotransmitter release. The syntaxin channel interaction may also support a novel signaling function, to modulate Ca(2+) channels according to the state of the associated release machinery (Bezprozvanny et al., 1995; Wiser et al., 1996; see also Mastrogiacomo et al., 1994). Here we report that syntaxin 1A (syn1A) coexpressed with N-type channels in Xenopus oocytes greatly promoted slow inactivation gating, but had little or no effect on the onset of and recovery from fast inactivation. Accordingly, the effectiveness of syntaxin depended strongly on voltage protocol. Slow inactivation was found for N-type channels even in the absence of syntaxin and could be distinguished from fast inactivation on the basis of its slow kinetics, distinct voltage dependence (voltage-independent at potentials higher than the level of half-inactivation), and temperature independence (Q(10), approximately 0.8). Trains of action potential-like stimuli were more effective than steady depolarizations in stabilizing the slowly inactivated condition. Agents that stimulate protein kinase C decreased the inhibitory effect of syntaxin on N-type channels. Application of BoNtC1 to cleave syntaxin sharply attenuated the modulatory effects on Ca(2+) channel gating, consistent with structural analysis of syntaxin modulation, supporting use of this toxin to test for the impact of syntaxin on Ca(2+) influx in nerve terminals. PMID- 10844005 TI - Syntaxin modulation of calcium channels in cortical synaptosomes as revealed by botulinum toxin C1. AB - When the presynaptic membrane protein syntaxin is coexpressed in Xenopus oocytes with N- or P/Q-type Ca(2+) channels, it promotes their inactivation (Bezprozvanny et al., 1995; Wiser et al., 1996, 1999; Degtiar et al., 2000) (I. B. Bezprozvanny, P. Zhong, R. H. Scheller, and R. W. Tsien, unpublished observations). These findings led to the hypothesis that syntaxin influences Ca(2+) channel function in presynaptic endings, in a reversal of the conventional flow of information from Ca(2+) channels to the release machinery. We examined this effect in isolated mammalian nerve terminals (synaptosomes). Botulinum neurotoxin type C1 (BoNtC1), which cleaves syntaxin, was applied to rat neocortical synaptosomes at concentrations that completely blocked neurotransmitter release. This treatment altered the pattern of Ca(2+) entry monitored with fura-2. Whereas the initial Ca(2+) rise induced by depolarization with K(+)-rich solution was unchanged, late Ca(2+) entry was strongly augmented by syntaxin cleavage. Similar results were obtained when Ca(2+) influx arose from repetitive firing induced by the K(+)-channel blocker 4-aminopyridine. Cleavage of vesicle-associated membrane protein with BoNtD or SNAP-25 with BoNtE failed to produce a significant change in Ca(2+) entry. The BoNtC1-induced alteration in Ca(2+) signaling was specific to voltage-gated Ca(2+) channels, not Ca(2+) extrusion or buffering, and it involved N-, P/Q- and R-type channels, the high voltage-activated channels most intimately associated with presynaptic release machinery. The modulatory effect of syntaxin was not immediately manifest when synaptosomes had been K(+)-predepolarized in the absence of external Ca(2+), but developed with a delay after admission of Ca(2+), suggesting that vesicular turnover may be necessary to make syntaxin available for its stabilizing effect on Ca(2+) channel inactivation. PMID- 10844006 TI - Opioid modulation of recurrent excitation in the hippocampal dentate gyrus. AB - kappa opioid receptor activation inhibits granule cell-mediated excitatory neurotransmission in the hippocampal formation via a decrease in glutamate release from both perforant path and mossy fiber terminals. We now report a third, anatomically and pharmacologically distinct site of such kappa opioid inhibition within the hippocampus. Granule cell population responses to selective stimulation of an excitatory hilar pathway were decreased by the kappa(1) opioid receptor agonist U69,593, an effect blocked by the kappa(1) antagonist norbinaltorphimine. U69,593 also inhibited hilar path induced long-term potentiation (LTP) of granule cell responses. LTP in this pathway was also blocked by the NMDA receptor antagonist d-2-amino-5-phosphonovalerate, unlike granule cell mossy fiber LTP in CA3. The kappa opioid peptide dynorphin is present in hilar mossy fiber collaterals. Ultrastructural analysis of these collaterals demonstrated dynorphin-containing vesicles in asymmetric synapses formed between axon terminals and granule cell dendrites, suggesting direct granule cell-granule cell connections. Evoked release of endogenous dynorphin within the hilus was effective in reducing hilar excitation of granule cells, although this release, in contrast to the release of dynorphin in the dentate molecular layer, was not dependent on L-type calcium channels. No hilar path excitation was observed in the absence of bicuculline, suggesting a strong GABA(A)-mediated inhibition of this pathway. However, hilar path activity could be seen after LTP, with or without bicuculline. Thus, kappa opioids can inhibit granule cell recurrent excitation, likely via effects on excitatory mossy fiber collaterals. Such collaterals are thought to be important in mediating temporal lobe epilepsy. PMID- 10844007 TI - Neuroprotective effects of creatine in a transgenic mouse model of Huntington's disease. AB - Huntington's disease (HD) is a progressive neurodegenerative illness for which there is no effective therapy. We examined whether creatine, which may exert neuroprotective effects by increasing phosphocreatine levels or by stabilizing the mitochondrial permeability transition, has beneficial effects in a transgenic mouse model of HD (line 6/2). Dietary creatine supplementation significantly improved survival, slowed the development of brain atrophy, and delayed atrophy of striatal neurons and the formation of huntingtin-positive aggregates in R6/2 mice. Body weight and motor performance on the rotarod test were significantly improved in creatine-supplemented R6/2 mice, whereas the onset of diabetes was markedly delayed. Nuclear magnetic resonance spectroscopy showed that creatine supplementation significantly increased brain creatine concentrations and delayed decreases in N-acetylaspartate concentrations. These results support a role of metabolic dysfunction in a transgenic mouse model of HD and suggest a novel therapeutic strategy to slow the pathological process. PMID- 10844008 TI - Inhibition of caspase-1-like activity by Ac-Tyr-Val-Ala-Asp-chloromethyl ketone induces long-lasting neuroprotection in cerebral ischemia through apoptosis reduction and decrease of proinflammatory cytokines. AB - Broad spectrum caspase inhibitors have been found to reduce neurodegeneration caused by cerebral ischemia. We studied whether blockade of group I caspases, mainly caspase-1, using the inhibitor Ac-YVAD.cmk reduced infarct volume and produced prolonged neuroprotection. Ac-YVAD.cmk (300 ng/rat) was injected intracerebroventricularly 10 min after permanent middle cerebral artery occlusion in the rat. Drug treatment induced a significant reduction of infarct volume not only 24 hr after ischemia (total damage, percentage of hemisphere volume: control, 41.1 +/- 2.3%; treated, 26.5 +/- 2.1%; p < 0.05) but also 6 d later (total damage: control, 30.6 +/- 2.2%; treated, 23.0 +/- 2.2%; p < 0.05). Ac YVAD. cmk treatment resulted in a reduction not only of caspase-1 (control, 100 +/- 20.3%; treated, 3.4 +/- 10.4%; p < 0.01) but also of caspase-3 (control, 100 +/- 30.3%; treated, 13.2 +/- 9.5%; p < 0.05) activity at 24 hr and led to a parallel decrease of apoptosis as measured by nucleosome quantitation (control, 100 +/- 11.8%; treated, 47 +/- 5.9%; p < 0.05). Six days after treatment no differences in these parameters could be detected between control and treated animals. Likewise, brain levels of the proinflammatory cytokines IL-1beta and TNF alpha were reduced at 24 hr (39.5 +/- 23.7 and 51.9 +/- 10.3% of control, respectively) but not at 6 d. Other cytokines, IL-10, MCP-1, MIP-2, and the gaseous mediator nitric oxide, were not modified by the treatment. These findings indicate that blockade of caspase-1-like activity induces a long-lasting neuroprotective effect that, in our experimental conditions, takes place in the early stages of damage progression. Finally, this effect is achieved by interfering with both apoptotic and inflammatory mechanisms. PMID- 10844009 TI - Dopamine-deficient mice are hypersensitive to dopamine receptor agonists. AB - Dopamine-deficient (DA-/-) mice were created by targeted inactivation of the tyrosine hydroxylase gene in dopaminergic neurons. The locomotor activity response of these mutants to dopamine D1 or D2 receptor agonists and l-3,4 dihydroxyphenylalanine (l-DOPA) was 3- to 13-fold greater than the response elicited from wild-type mice. The enhanced sensitivity of DA-/- mice to agonists was independent of changes in steady-state levels of dopamine receptors and the presynaptic dopamine transporter as measured by ligand binding. The acute behavioral response of DA-/- mice to a dopamine D1 receptor agonist was correlated with c-fos induction in the striatum, a brain nucleus that receives dense dopaminergic input. Chronic replacement of dopamine to DA-/- mice by repeated l-DOPA administration over 4 d relieved the hypersensitivity of DA-/- mutants in terms of induction of both locomotion and striatal c-fos expression. The results suggest that the chronic presence of dopaminergic neurotransmission is required to dampen the intracellular signaling response of striatal neurons. PMID- 10844010 TI - Probing fundamental aspects of synaptic transmission with strontium. AB - Strontium is capable of supporting synaptic transmission, but release is dramatically different from that evoked in calcium. By measuring presynaptic strontium levels, we gain insight into the actions of strontium, which has implications for the identification of molecules involved in different aspects of synaptic transmission. We examined presynaptic divalent levels and synaptic release at the granule cell to stellate cell synapse in mouse cerebellar slices. We find that the prolonged duration of release and paired-pulse facilitation in the presence of strontium can be accounted for by the slower removal of strontium from the presynaptic terminal. Phasic and delayed release are both driven by strontium less effectively than by calcium, indicating that a heightened sensitivity to strontium is not a feature of the binding sites involved in facilitation and delayed release. We also find that the cooperativity for phasic release is 1.7 for strontium compared with 3.2 for calcium, suggesting that differential binding may help to identify the calcium sensor involved in phasic release. PMID- 10844011 TI - Prolonged synaptic currents and glutamate spillover at the parallel fiber to stellate cell synapse. AB - Although neurons often fire in bursts, most of what is known about glutamate signaling and postsynaptic receptor activation is based on experiments using single stimuli. Here we examine the activation of ionotropic glutamate receptors by bursts at the parallel fiber to stellate cell synapse. We show that brief stimulus trains generate prolonged AMPA receptor (AMPAR)- and NMDA receptor (NMDAR)-mediated EPSCs recorded in whole-cell voltage clamp. These EPSCs contrast with the rapid AMPAR-mediated EPSC evoked by a single stimulus. The prolonged AMPAR-mediated EPSC is promoted by high-frequency and high-intensity trains and can persist for hundreds of milliseconds. This EPSC is also increased by l-trans 2,4-PDC, an inhibitor of glutamate transporters, suggesting that these transporters usually limit the synaptic response to trains. These prolonged EPSCs reflect both receptor properties and a long-lasting glutamate signal. In addition, several experiments demonstrate that glutamate spillover can contribute to receptor activation. First, imaging stimulus-evoked changes in presynaptic calcium establishes that distinct parallel fiber bands can be activated. Second, activation of parallel fibers that do not directly synapse onto a given stellate cell can evoke indirect AMPAR- and NMDAR-mediated EPSCs in that cell. Third, experiments using the use-dependent NMDAR blocker MK-801 show that these indirect EPSCs reflect glutamate spillover in response to trains. Together, these findings indicate that stimulus trains can generate a sustained and widespread glutamate signal that can in turn evoke large and prolonged EPSCs mediated by ionotropic glutamate receptors. These synaptic properties may have important functional consequences for stellate cell firing. PMID- 10844012 TI - Extensive sprouting of sensory afferents and hyperalgesia induced by conditional expression of nerve growth factor in the adult spinal cord. AB - Genetic transfer of growth-promoting molecules was proposed as a potential strategy to modify the nonpermissive nature of the adult CNS to induce axonal regeneration. To evaluate whether overexpression of neurotrophins or cellular adhesion molecules would effect axonal plasticity, adenoviruses encoding fibroblast growth factor-2 (FGF-2/Adts), nerve growth factor (NGF/Adts), neurotrophin-3, and the cell adhesion molecules N-cadherin and L1 were injected into the dorsal horn of the adult spinal cord. Transgene expression was primarily localized to astrocytes in the dorsal horn and motor neurons within the ventral horn. Overexpression of these factors, with the exception of NGF/Adts, failed to increase axonal sprouting. Eight days after NGF/Adts injections, axonal sprouting within the dorsal horn was apparent, and after 4 weeks, extensive spouting was observed throughout the entire dorsal horn, extending into the ventral horn and the white matter of the lateral funiculus. These axons were identified primarily as a subpopulation of nociceptive fibers expressing calcitonin gene-related peptide and substance-P. Behavioral analysis revealed thermal hyperalgesia and perturbation of accurate paw placement on grid-walking tasks for both FGF-2- and NGF-treated animals. These results indicate that the administration of growth promoting molecules can induce robust axonal plasticity of normal adult primary sensory neurons into areas of transgene expression, causing significant alterations in behavioral responses. This observation also indicates that gene transfer protocols that aim to reconstruct diseased or injured pathways should also be designed to prevent the sprouting of the normal circuitry from adjacent unaffected neurons. PMID- 10844013 TI - Inhibition of the cAMP pathway decreases early long-term potentiation at CA1 hippocampal synapses. AB - Long-term potentiation (LTP) has several different phases, and there is general agreement that the late phase of LTP requires the activation of adenylyl cyclase (AC) and cAMP-dependent protein kinase (PKA). In contrast, several studies indicate that the early LTP is not affected by interfering with the cAMP pathway. We have further tested the role of the cAMP pathway in early LTP using several types of inhibitors. Bath application of the PKA inhibitor H89 suppressed the early LTP induced by a single tetanus. Similarly, the LTP induced by a pairing protocol was decreased by postsynaptic intracellular perfusion of the peptide PKA inhibitor PKI(6-22) amide. The decrease of LTP produced by these inhibitors was evident immediately after induction. These results indicate that PKA is important in early LTP, that its locus of action is postsynaptic, and that it does not act merely by enhancing the depolarization required for LTP induction. The failure of some other inhibitors of the cAMP pathway to affect the early phase of LTP might be attributable to the saturation of some step in the cAMP pathway during a tetanus. In agreement with this hypothesis we found that application of the AC inhibitor SQ 22536 by itself did not affect the early phase of LTP, but did produce a reduction if the cAMP pathway was already attenuated by the PKA inhibitor H89. Our analysis of the results of genetic modifications of the cAMP pathway, especially the work on AC knock-outs, indicates that the genetic data are generally consistent with the pharmacological results showing the importance of this pathway in early LTP. PMID- 10844014 TI - In CA1 pyramidal neurons of the hippocampus protein kinase C regulates calcium dependent inactivation of NMDA receptors. AB - The NMDA subtype of the glutamate-gated channel exhibits a high permeability to Ca(2+). The influx of Ca(2+) through NMDA channels is limited by a rapid and Ca(2+)/calmodulin (CaM)-dependent inactivation that results from a competitive displacement of cytoskeleton-binding proteins from the NR1 subunit of the receptor by Ca(2+)/CaM (Zhang et al., 1998; Krupp et al., 1999). The C terminal of this subunit can be phosphorylated by protein kinase C (PKC) (Tingley et al., 1993). The present study sought to investigate whether PKC regulates Ca(2+) dependent inactivation of the NMDA channel in hippocampal neurons. Activation of endogenous PKC by 4beta-phorbol 12-myristate 13-acetate enhanced peak (I(p)) and depressed steady-state (I(ss)) NMDA-evoked currents, resulting in a reduction in the ratio of these currents (I(ss)/I(p)). We demonstrated previously that PKC activity enhances I(P) via a sequential activation of the focal adhesion kinase cell adhesion kinase beta/proline-rich tyrosine kinase 2 (CAKbeta/Pyk2) and the nonreceptor tyrosine kinase Src (Huang et al., 1999; Lu et al., 1999). Here, we report that the PKC-induced depression of I(ss) is unrelated to the PKC/CAKbeta/Src-signaling pathway but depends on the concentration of extracellular Ca(2+). Intracellular applications of CaM reduced I(ss)/I(p) and occluded the Ca(2+)-dependent effect of phorbol esters on I(ss.) Moreover, increasing the concentration of intracellular Ca(2+) buffer or intracellular application of the inhibitory CaM-binding peptide (KY9) greatly reduced the phorbol ester-induced depression of I(ss). Taken together, these results suggest that PKC enhances Ca(2+)/CaM-dependent inactivation of the NMDA channel, most likely because of a phosphorylation-dependent regulation of interactions between receptor subunits, CaM, and other postsynaptic density proteins. PMID- 10844015 TI - Functional architecture of synapses in the inner retina: segregation of visual signals by stratification of bipolar cell axon terminals. AB - We correlated the morphology of salamander bipolar cells with characteristics of their light responses, recorded under voltage-clamp conditions. Twelve types of bipolar cells were identified, each displaying a unique morphology and level(s) of axon terminal stratification in the inner plexiform layer (IPL) and exhibiting light responses that differed with respect to polarity, kinetics, the relative strengths of rod and cone inputs, and characteristics of spontaneous EPSCs (sEPSCs) and IPSCs. In addition to the well known segregation of visual information into ON and OFF channels along the depth of the IPL, we found an overlying mapping of spectral information in this same dimension, with cone signals being transmitted predominantly to the central IPL and rod signals being sent predominantly to the margins of the IPL. The kinetics of bipolar cell responses correlated with this segregation of ON and OFF and of rod and cone information in the IPL. At light offset, rod-dominated cells displayed larger slow cationic current tails and smaller rapid overshoot responses than did cone dominated cells. sEPSCs were generally absent in depolarizing bipolar cells but present in all hyperpolarizing bipolar cells (HBCs) and larger in rod-dominated HBCs than in cone-dominated HBCs. Inhibitory chloride currents, elicited both at light onset and light offset, tended to be larger for cone-dominated cells than for rod-dominated cells. This orderly segregation of visual signals along the depth of the IPL simplifies the integration of visual information in the retina, and it begins a chain of parallel processing in the visual system. PMID- 10844016 TI - Regulation of the on bipolar cell mGluR6 pathway by Ca2+. AB - Glutamate produces a hyperpolarizing synaptic potential in On bipolar cells by binding to the metabotropic glutamate receptor mGluR6, leading to closure of a cation channel. Here it is demonstrated that this cation channel is regulated by intracellular Ca(2+). Glutamate-evoked currents were recorded from On bipolar cells in light-adapted salamander retinal slices in the presence of 2 mm external Ca(2+). When glutamate was applied almost continuously, interrupted only briefly to measure the size of the response, the glutamate response remained robust. However, currents elicited by intermittent and brief applications of glutamate exhibited time-dependent run down. Run down of the glutamate response was also voltage dependent, because it was accelerated by membrane hyperpolarization. Run down was triggered, at least in part, by a rise in intracellular Ca(2+); measured as a function of time or voltage, it was attenuated by intracellular buffering of Ca(2+) with BAPTA or by omitting Ca(2+) from the bathing solution. Current voltage measurements demonstrated that Ca(2+) induced run down of the glutamate response by downregulating cation channel function, rather than by preventing closure of the channel by glutamate and mGluR6. A major source of the Ca(2+) that mediated this inhibition is the cation channel itself, which was found to be permeable to Ca(2+), accounting for the use dependence of the run down. These results suggest that Ca(2+) influx through the cation channel during background illumination could provide a signal to close the cation channel and repolarize the membrane toward its dark potential, an adaptive mechanism for coping with changes in ambient light. PMID- 10844017 TI - Regulation of phosphorylation of the GluR1 AMPA receptor in the neostriatum by dopamine and psychostimulants in vivo. AB - The activation of cAMP-dependent protein kinase regulates the physiological activity of AMPA-type glutamate receptors. In this study, phosphorylation of the AMPA receptor subunit GluR1 at Ser(845) was increased in neostriatal slices by activation of D1-type dopamine receptors and by inhibitors of protein phosphatase 1/protein phosphatase 2A. In contrast, Ser(831), a residue which, when phosphorylated by protein kinase C or calcium/calmodulin-dependent kinase II, increases AMPA receptor channel conductance, was unaffected by either D1 or D2 receptor agonists in neostriatal slices. The phosphorylation of Ser(845), but not Ser(831), was strongly increased in neostriatum in vivo in response to the psychostimulants cocaine and methamphetamine. The effects of dopamine and psychostimulants on the phosphorylation of GluR1 were attenuated in dopamine and cAMP-regulated phosphoprotein M(r) 32 kDa (DARPP-32) knock-out mice. These results identify DARPP-32 and AMPA-type glutamate receptors as likely essential cellular effectors for psychostimulant actions. PMID- 10844018 TI - Endogenous regulator of G-protein signaling proteins modify N-type calcium channel modulation in rat sympathetic neurons. AB - Experiments using heterologous overexpression indicate that regulator of G protein signaling (RGS) proteins play important roles in Gbetagamma-mediated ion channel modulation. However, the roles subserved by endogenous RGS proteins have not been extensively examined because tools for functionally inhibiting natively expressed RGS proteins are lacking. To address this void, we used a strategy in which Galpha(oA) was rendered insensitive to pertussis toxin (PTX) and RGS proteins by site-directed mutagenesis. Either PTX-insensitive (PTX-i) or both PTX and RGS-insensitive (PTX/RGS-i) mutants of Galpha(oA) were expressed along with Gbeta(1) and Ggamma(2) subunits in rat sympathetic neurons. After overnight treatment with PTX to suppress natively expressed Galpha subunits, voltage dependent Ca(2+) current inhibition by norepinephrine (NE) (10 microm) was reconstituted in neurons expressing either PTX-i or PTX/RGS-i Galpha(oA). When compared with neurons expressing PTX-i Galpha(oA), the steady-state concentration response relationships for NE-induced Ca(2+) current inhibition were shifted to lower concentrations in neurons expressing PTX/RGS-i Galpha(oA). In addition to an increase in agonist potency, the expression of PTX/RGS-i Galpha(oA) dramatically retarded the current recovery after agonist removal. Interestingly, the alteration in current recovery was accompanied by a slowing in the onset of current inhibition. Together, our data suggest that endogenous RGS proteins contribute to membrane-delimited Ca(2+) channel modulation by regulating agonist potency and kinetics of G-protein-mediated signaling in neuronal cells. PMID- 10844019 TI - Presynaptic protein kinase activity supports long-term potentiation at synapses between individual hippocampal neurons. AB - Simultaneous microelectrode recording from two individual synaptically connected neurons enables the direct analysis of synaptic transmission and plasticity at a minimal synaptic connection. We have recorded from pairs of CA3 pyramidal neurons in organotypic hippocampal slices to examine the properties of long-term potentiation (LTP) at such minimal connections. LTP in minimal connections was found to be identical to the NMDA-dependent LTP expressed by CA3-CA1 synapses, demonstrating this system provides a good model for the study of the mechanisms of LTP expression. The LTP at minimal synaptic connections does not behave as a simple increase in transmitter release probability, because the amplitude of unitary EPSCs can increase several-fold, unlike what is observed when release probability is increased by raising extracellular calcium. Taking advantage of the relatively short axon connecting neighboring CA3 neurons, we found it feasible to introduce pharmacological agents to the interior of presynaptic terminals by injection into the presynaptic soma and have used this technique to investigate presynaptic effects on basal transmission and LTP. Presynaptic injection of nicotinamide reduced basal transmission, but LTP in these pairs was essentially normal. In contrast, presynaptic injection of H-7 significantly depressed LTP but not basal transmission, indicating a specific role of presynaptic protein kinases in LTP. These results demonstrate that pharmacological agents can be directly introduced into the presynaptic cell and that a purely presynaptic perturbation can alter this plasticity. PMID- 10844020 TI - Activation of mitogen-activated protein kinases after transient forebrain ischemia in gerbil hippocampus. AB - We investigated the expression, activation, and distribution of c-Jun N-terminal kinases (JNKs), p38 mitogen-activated protein kinases (p38s) and extracellular signal-regulated kinases (ERKs) using Western blotting and immunohistochemistry in gerbil hippocampus after transient forebrain ischemia to clarify the role of these kinases in delayed neuronal death (DND) in the CA1 subfield. Immunoblot analysis demonstrated that activities of JNK, p38, and ERK in whole hippocampus were increased after 5 min of global ischemia. We used an immunohistochemical study to elucidate the temporal and spatial expression of these kinases after transient global ischemia. The immunohistochemical study showed that active JNK and p38 immunoreactivities were enhanced at 15 min of reperfusion and then gradually reduced and disappeared in the hippocampal CA1 region. On the other hand, in CA3 neurons, active JNK and p38 immunoreactivities were enhanced at 15 min of reperfusion and peaked at 6 hr of reperfusion and then gradually reduced but was continuously detected 72 hr after ischemia. Active ERK immunoreactivity was observed transiently in CA3 fibers and dentate gyrus. Pretreatment with SB203580, a p38 inhibitor, but not with PD98059, an ERK kinase 1/2 inhibitor, reduced ischemic cell death in the CA1 region after transient global ischemia by inhibiting the activity of p38. These findings indicate that the p38 pathway may play an important role in DND during brain ischemia in gerbil. Components of the pathway are important target molecules for clarifying the mechanism of neuronal death. PMID- 10844021 TI - The L1-type cell adhesion molecule neuroglian influences the stability of neural ankyrin in the Drosophila embryo but not its axonal localization. AB - Ankyrins are linker proteins, which connect various membrane proteins, including members of the L1 family of neural cell adhesion molecules, with the submembranous actin-spectrin skeleton. Here we report the cloning and characterization of a second, novel Drosophila ankyrin gene (Dank2) that appears to be the result of a gene duplication event during arthropod evolution. The Drosophila L1-type protein neuroglian interacts with products from both Drosophila ankyrin genes. Whereas the previously described ankyrin gene is ubiquitously expressed during embryogenesis, the expression of Dank2 is restricted to the nervous system in the Drosophila embryo. The absence of neuroglian protein in a neuroglian null mutant line causes decreased levels of Dank2 protein in most neuronal cells. This suggests that neuroglian is important for the stability of Dank2 protein. However, neuroglian is not required for Dank2 axonal localization. In temperature-sensitive neuroglian mutants in which neuroglian protein is mislocated at the restrictive temperature to an intracellular location in the neuronal soma, Dank2 protein can still be detected along embryonic nerve tracts. PMID- 10844022 TI - Interaction of the postsynaptic density-95/guanylate kinase domain-associated protein complex with a light chain of myosin-V and dynein. AB - NMDA receptors interact directly with postsynaptic density-95 (PSD-95), a scaffold protein that organizes a cytoskeletal- signaling complex at the postsynaptic membrane. The molecular mechanism by which the PSD-95-based protein complex is trafficked to the postsynaptic site is unknown but presumably involves specific motor proteins. Here we demonstrate a direct interaction between the PSD 95-associated protein guanylate kinase domain-associated protein (GKAP) and dynein light chain (DLC), a light chain subunit shared by myosin-V (an actin based motor) and cytoplasmic dynein (a microtubule-based motor). A yeast two hybrid screen with GKAP isolated DLC2, a novel protein 93% identical to the previously cloned 8 kDa dynein light chain (DLC1). A complex containing PSD-95, GKAP, DLC, and myosin-V can be immunoprecipitated from rat brain extracts. DLC colocalizes with PSD-95 and F-actin in dendritic spines of cultured neurons and is enriched in biochemical purifications of PSD. Immunogold electron microscopy reveals a concentration of DLC in the postsynaptic compartment of asymmetric synapses of brain in which it is associated with the PSD and the spine apparatus. We discuss the possibility that the GKAP/DLC interaction may be involved in trafficking of the PSD-95 complex by motor proteins. PMID- 10844023 TI - Phosphorylated syntaxin 1 is localized to discrete domains along a subset of axons. AB - Syntaxin 1 is a SNARE protein that plays a central role in synaptic vesicle (SV) exocytosis. We generated an antibody that specifically recognizes a casein kinase II-mediated phosphorylation on serine-14 of syntaxin 1. In this report we show that this phosphorylation occurs in vivo and is developmentally regulated in the rat brain, rising to a level of 40% of the total syntaxin in adult animals. Phosphorylated syntaxin is preferentially associated with SNAP-25 and localizes to discrete domains of the axonal plasma membrane that do not colocalize with pools of synaptic vesicles. These phosphosyntaxin domains may define fusion sites for a novel class of vesicles outside classical active zones. PMID- 10844024 TI - Postsynaptic scaffolds of excitatory and inhibitory synapses in hippocampal neurons: maintenance of core components independent of actin filaments and microtubules. AB - The mechanisms responsible for anchoring molecular components of postsynaptic specializations in the mammalian brain are not well understood but are presumed to involve associations with cytoskeletal elements. Here we build on previous studies of neurotransmitter receptors (Allison et al., 1998) to analyze the modes of attachment of scaffolding and signal transducing proteins of both glutamate and GABA postsynaptic sites to either the microtubule or microfilament cytoskeleton. Hippocampal pyramidal neurons in culture were treated with latrunculin A to depolymerize actin, with vincristine to depolymerize microtubules, or with Triton X-100 to extract soluble proteins. The synaptic clustering of PSD-95, a putative NMDA receptor anchoring protein and a core component of the postsynaptic density (PSD), was unaffected by actin depolymerization, microtubule depolymerization, or detergent extraction. The same was largely true for GKAP, a PSD-95-interacting protein. In contrast, the synaptic clustering of Ca(2+)/calmodulin-dependent protein kinase II (CaMKII)alpha, another core component of the PSD, was completely dependent on an intact actin cytoskeleton and was partially disrupted by detergent. Drebrin and alpha-actinin-2, actin-binding proteins concentrated in spines, were also dependent on F-actin for synaptic localization but were unaffected by detergent extraction. Surprisingly, the subcellular distributions of the inhibitory synaptic proteins GABA(A)R and gephyrin, which has a tubulin-binding motif, were unaffected by depolymerization of microtubules or actin or by detergent extraction. These studies reveal an unsuspected heterogeneity in the modes of attachment of postsynaptic proteins to the cytoskeleton and support the idea that PSD-95 and gephyrin may be core scaffolding components independent of the actin or tubulin cytoskeleton. PMID- 10844025 TI - Chronic heroin self-administration desensitizes mu opioid receptor-activated G proteins in specific regions of rat brain. AB - In previous studies from our laboratory, chronic noncontingent morphine administration decreased mu opioid receptor-activated G-proteins in specific brainstem nuclei. In the present study, mu opioid receptor binding and receptor activated G-proteins were examined after chronic heroin self-administration. Rats were trained to self-administer intravenous heroin for up to 39 d, achieving heroin intake up to 366 mg. kg(-1). d(-1). mu opioid-stimulated [(35)S]GTPgammaS and [(3)H]naloxone autoradiography were performed in adjacent brain sections. Agonist-stimulated [(35)S]GTPgammaS autoradiography also examined other G-protein coupled receptors, including delta opioid, ORL-1, GABA(B), adenosine A(1), cannabinoid, and 5-HT(1A). In brains from heroin self-administering rats, decreased mu opioid-stimulated [(35)S]GTPgammaS binding was observed in periaqueductal gray, locus coeruleus, lateral parabrachial nucleus, and commissural nucleus tractus solitarius, as previously observed in chronic morphine-treated animals. In addition, decreased mu opioid-stimulated [(35)S]GTPgammaS binding was found in thalamus and amygdala after heroin self administration. Despite this decrease in mu-activated G-proteins, [(3)H]naloxone binding demonstrated increased mu opioid receptor binding in several brain regions after heroin self-administration, and there was a significant decrease in mu receptor G-protein efficiency as expressed as a ratio between agonist activated G-proteins and mu receptor binding. No effects on agonist-stimulated [(35)S]GTPgammaS binding were found for any other receptor examined. The effect of chronic heroin self-administration to decrease mu-stimulated [(35)S]GTPgammaS binding varied between regions and was highest in brainstem and lowest in the cortex and striatum. These results not only provide potential neuronal mechanisms that may contribute to opioid tolerance and dependence, but also may explain why various chronic effects of opioids develop to different degrees. PMID- 10844026 TI - The MAPK/ERK cascade targets both Elk-1 and cAMP response element-binding protein to control long-term potentiation-dependent gene expression in the dentate gyrus in vivo. AB - The mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) signaling cascade contributes to synaptic plasticity and to long-term memory formation, yet whether MAPK/ERK controls activity-dependent gene expression critical for long-lasting changes at the synapse and what the events underlying transduction of the signal are remain uncertain. Here we show that induction of long-term potentiation (LTP) in the dentate gyrus in vivo leads to rapid phosphorylation and nuclear translocation of MAPK/ERK. Following a similar time course, the two downstream transcriptional targets of MAPK/ERK, cAMP response element-binding protein (CREB) and the ternary complex factor Elk-1, a key transcriptional-regulator of serum response element (SRE)-driven gene expression, were hyperphosphorylated and the immediate early gene zif268 was upregulated. The mRNA encoding MAP kinase phosphatase MKP-1 was upregulated at the time point when MAPK/ERK phosphorylation had returned to basal levels, suggesting a negative feedback loop to regulate deactivation of MAPK/ERK. We also show that inhibition of the MAPK/ERK cascade by the MAPK kinase MEK inhibitor SL327 prevented CREB and Elk-1 phosphorylation, and LTP-dependent gene induction, resulting in rapidly decaying LTP. In conclusion, we suggest that Elk-1 forms an important link in the MAP kinase pathway to transduce signals from the cell surface to the nucleus to activate the genetic machinery necessary for the maintenance of synaptic plasticity in the dentate gyrus. Thus, MAPK/ERK activation is required for LTP-dependent transcriptional regulation and we suggest this is regulated by two parallel signaling pathways, the MAPK/ERK-Elk-1 pathway targeting SRE and the MAPK/ERK-CREB pathway targeting CRE. PMID- 10844027 TI - C-Terminal truncation of NR2A subunits impairs synaptic but not extrasynaptic localization of NMDA receptors. AB - NMDA receptors interact via the extended intracellular C-terminal domain of the NR2 subunits with constituents of the postsynaptic density for purposes of retention, clustering, and functional regulation at central excitatory synapses. To examine the role of the C-terminal domain of NR2A in the synaptic localization and function of NR2A-containing NMDA receptors in hippocampal Schaffer collateral CA1 pyramidal cell synapses, we analyzed mice which express NR2A only in its C terminally truncated form. In CA1 cell somata, the levels, activation, and deactivation kinetics of extrasynaptic NMDA receptor channels were comparable in wild-type and mutant NR2A(Delta)(C/)(Delta)(C) mice. At CA1 cell synapses, however, the truncated receptors were less concentrated than their full-length counterparts, as indicated by immunodetection in cultured neurons, synaptosomes, and postsynaptic densities. In the mutant, the NMDA component of evoked EPSCs was reduced in a developmentally progressing manner and was even more reduced in miniature EPSCs (mEPSCs) elicited by spontaneous glutamate release. Moreover, pharmacologically isolated NMDA currents evoked by synaptic stimulation had longer latencies and displayed slower rise and decay times, even in the presence of an NR2B-specific antagonist. These data strongly suggest that the C-terminal domain of NR2A subunits is important for the precise synaptic arrangement of NMDA receptors. PMID- 10844028 TI - Potentiation of NMDA receptor function by the serine protease thrombin. AB - Although serine proteases and their receptors are best known for their role in blood coagulation and fibrinolysis, the CNS expresses many components of an extracellular protease signaling system including the protease-activated receptor 1 (PAR1), for which thrombin is the most effective activator. In this report we show that activation of PAR1 potentiates hippocampal NMDA receptor responses in CA1 pyramidal cells by 2.07 +/- 0.27-fold (mean +/- SEM). Potentiation of neuronal NMDA receptor responses by thrombin can be blocked by thrombin and a protein kinase inhibitor, and the effects of thrombin can be mimicked by a peptide agonist (SFLLRN) that activates PAR1. Potentiation of the NMDA receptor by thrombin in hippocampal neurons is significantly attenuated in mice lacking PAR1. Although high concentrations of thrombin can directly cleave both native and recombinant NR1 subunits, the thrombin-induced potentiation we observe is independent of NMDA receptor cleavage. Activation of recombinant PAR1 also potentiates recombinant NR1/NR2A (1.7 +/- 0.06-fold) and NR1/NR2B (1.41 +/- 0.11 fold) receptor function but not NR1/NR2C or NR1/NR2D receptor responses. PAR1 mediated potentiation of recombinant NR1/NR2A receptors occurred after activation with as little as 300 pm thrombin. These data raise the intriguing possibility that potentiation of neuronal NMDA receptor function after entry of thrombin or other serine proteases into brain parenchyma during intracerebral hemorrhage or extravasation of plasma proteins during blood-brain barrier breakdown may exacerbate glutamate-mediated cell death and possibly participate in post traumatic seizure. Furthermore, the ability of neuronal protease signaling to control NMDA receptor function may also have roles in normal brain development. PMID- 10844029 TI - Functional interactions between Drosophila bHLH/PAS, Sox, and POU transcription factors regulate CNS midline expression of the slit gene. AB - During Drosophila embryogenesis the CNS midline cells have organizing activities that are required for proper elaboration of the axon scaffold and differentiation of neighboring neuroectodermal and mesodermal cells. CNS midline development is dependent on Single-minded (Sim), a basic-helix-loop-helix (bHLH)-PAS transcription factor. We show here that Fish-hook (Fish), a Sox HMG domain protein, and Drifter (Dfr), a POU domain protein, act in concert with Single minded to control midline gene expression. single-minded, fish-hook, and drifter are all expressed in developing midline cells, and both loss- and gain-of function assays revealed genetic interactions between these genes. The corresponding proteins bind to DNA sites present in a 1 kb midline enhancer from the slit gene and regulate the activity of this enhancer in cultured Drosophila Schneider line 2 cells. Fish-hook directly associates with the PAS domain of Single-minded and the POU domain of Drifter; the three proteins can together form a ternary complex in yeast. In addition, Fish can form homodimers and also associates with other bHLH-PAS and POU proteins. These results indicate that midline gene regulation involves the coordinate functions of three distinct types of transcription factors. Functional interactions between members of these protein families may be important for numerous developmental and physiological processes. PMID- 10844030 TI - In utero cocaine-induced dysfunction of dopamine D1 receptor signaling and abnormal differentiation of cerebral cortical neurons. AB - Monoamines modulate neuronal differentiation, and alteration of monoamine neurotransmission during development produces specific changes in neuronal structure, function, and pattern formation. We have previously observed that prenatal exposure to cocaine in a clinically relevant animal model produces increased length of pyramidal neuron dendrites in the anterior cingulate cortex (ACC) postnatally. We now report that cocaine administered intravenously to pregnant rabbits at gestational stages preceding and during cortical histogenesis results in the early onset of hypertrophic dendritic outgrowth in the embryonic ACC. Confocal microscopy of DiI-labeled neurons revealed that the atypical, tortuous dendritic profiles seen postnatally in ACC-cocaine neurons already are apparent in utero. No defects in neuronal growth were observed in visual cortex (VC), a region lacking prominent dopamine innervation. In striking correlation with our in vivo results, in vitro experiments revealed a significant enhancement of spontaneous process outgrowth of ACC neurons isolated from cocaine-exposed fetuses but no changes in neurons derived from visual cortex. The onset of modified growth in vivo is paralleled by reduced D(1A) receptor coupling to its G protein. These data suggest that the dynamic growth of neurons can be regulated by early neurotransmitter signaling in a selective fashion. Prenatal onset of defects in dopamine receptor signaling contributes to abnormal circuit formation and may underlie specific cognitive and behavioral dysfunction. PMID- 10844031 TI - Lens injury stimulates axon regeneration in the mature rat optic nerve. AB - In mature mammals, retinal ganglion cells (RGCs) are unable to regenerate their axons after optic nerve injury, and they soon undergo apoptotic cell death. However, a small puncture wound to the lens enhances RGC survival and enables these cells to regenerate their axons into the normally inhibitory environment of the optic nerve. Even when the optic nerve is intact, lens injury stimulates macrophage infiltration into the eye, Muller cell activation, and increased GAP 43 expression in ganglion cells across the entire retina. In contrast, axotomy, either alone or combined with intraocular injections that do not infringe on the lens, causes only a minimal change in GAP-43 expression in RGCs and a minimal activation of the other cell types. Combining nerve injury with lens puncture leads to an eightfold increase in RGC survival and a 100-fold increase in the number of axons regenerating beyond the crush site. Macrophage activation appears to play a key role, because intraocular injections of Zymosan, a yeast cell wall preparation, stimulated monocytes in the absence of lens injury and induced RGCs to regenerate their axons into the distal optic nerve. PMID- 10844032 TI - Cell cycle control of Schwann cell proliferation: role of cyclin-dependent kinase 2. AB - Schwann cell proliferation is regulated by multiple growth factors and axonal signals. However, the molecules that control growth arrest of Schwann cells are not well defined. Here we describe regulation of the cyclin-dependent kinase-2 (CDK2) protein, an enzyme that is necessary for the transition from G1 to S phase. Levels of CDK2 protein were elevated in proliferating Schwann cells cultured in serum and forskolin. However, when cells were grown with either serum free media or at high densities, CDK2 levels declined to low levels. The decrease in CDK2 levels was associated with growth arrest of Schwann cells. The modulation of CDK2 appears to be regulated at the transcriptional level, because CDK2 mRNA levels and its promoter activity both decline during cell cycle arrest. Furthermore, analysis of the CDK2 promoter suggests that Sp1 DNA binding sites are essential for maximal activation in Schwann cells. Together, these data suggest that CDK2 may represent a significant target of developmental signals that regulate Schwann cell proliferation and that this regulation is mediated, in part, through regulation of Sp1 transcriptional activity. PMID- 10844033 TI - Axonal regulation of Schwann cell proliferation and survival and the initial events of myelination requires PI 3-kinase activity. AB - In this report, we have investigated the signaling pathways that are activated by, and mediate the effects of, the neuregulins and axonal contact in Schwann cells. Phosphatidylinositol 3-kinase (PI 3-kinase) and mitogen-activated protein kinase kinase (MAPK kinase) are strongly activated in Schwann cells by glial growth factor (GGF), a soluble neuregulin, and by contact with neurite membranes; both kinase activities are also detected in Schwann cell-DRG neuron cocultures. Inhibition of the PI 3-kinase, but not the MAP kinase, pathway reversibly inhibited Schwann cell proliferation induced by GGF and neurites. Cultured Schwann cells undergo apoptosis after serum deprivation and can be rescued by GGF or contact with neurites; these survival effects were also blocked by inhibition of PI 3-kinase. Finally, we have examined the role of these signaling pathways in Schwann cell differentiation in cocultures. At early stages of coculture, inhibition of PI 3-kinase, but not MAPK kinase, blocked Schwann cell elongation and subsequent myelination but did not affect laminin deposition. Later, after Schwann cells established a one-to-one relationship with axons, inhibition of PI 3-kinase did not block myelin formation, but the myelin sheaths that formed were shorter, and the rate of myelin protein accumulation was markedly decreased. PI 3 kinase inhibition had no observable effect on the maintenance of myelin sheaths in mature myelinated cocultures. These results indicate that activation of PI 3 kinase by axonal factors, including the neuregulins, promotes Schwann cell proliferation and survival and implicate PI 3-kinase in the early events of myelination. PMID- 10844034 TI - Abnormal phrenic motoneuron activity and morphology in neonatal monoamine oxidase A-deficient transgenic mice: possible role of a serotonin excess. AB - In rodent neonates, the neurotransmitter serotonin (5-HT) modulates the activity of both the medullary respiratory rhythm generator and the cervical phrenic motoneurons. To determine whether 5-HT also contributes to the maturation of the respiratory network, experiments were conducted in vitro on the brainstem-spinal cord preparation of neonatal mice originating from the control strain (C3H) and the monoamine oxidase A-deficient strain, which has a brain perinatal 5-HT excess (Tg8). At birth, the Tg8 respiratory network is unable to generate a respiratory pattern as stable as that produced by the C3H network, and the modulation by 5-HT of the network activity present in C3H neonates is lacking in Tg8 neonates. In addition, the morphology of the phrenic motoneurons is altered in Tg8 neonates; the motoneuron dendritic tree loses the C3H bipolar aspect but exhibits an increased number of spines and varicosities. These abnormalities were prevented in Tg8 neonates by treating pregnant Tg8 dams with the 5-HT synthesis inhibitor p chlorophenylalanine or a 5-HT(2A) receptor antagonist but were induced in wild type neonates by treating C3H dams with a 5-HT(2A) receptor agonist. We conclude that 5-HT contributes, probably via 5-HT(2A) receptors, to the normal maturation of the respiratory network but alters it when present in excess. Disorders affecting 5-HT metabolism during gestation may therefore have deleterious effects on newborns. PMID- 10844035 TI - Distribution of corticosteroid receptors in the rhesus brain: relative absence of glucocorticoid receptors in the hippocampal formation. AB - Chronic stress has been associated with degenerative changes in the rodent and primate hippocampus, presumably mediated in part via neuronal glucocorticoid receptors (GRs). In the rat brain, GRs are widely distributed and are particularly dense in the hippocampus. The distribution of GRs in the primate brain, however, has not been fully characterized. In this study, we used in situ hybridization histochemistry and immunohistochemistry to map the distribution of GR mRNA and GR protein, respectively, in adult rhesus monkeys (Macaca mulatta). In contrast to its well established distribution in the rat brain, GR mRNA was only weakly detected in the dentate gyrus (DG) and Cornu Ammonis (CA) of the macaque hippocampus, whereas it was abundant in the pituitary (PIT), cerebellum (CBL), hypothalamic paraventricular nucleus (PVN), and, to a lesser extent, the neocortex. Immunohistochemical staining indicated a very low density of GR-like immunoreactive cells within the macaque hippocampal formation in contrast to the high density observed within the PVN, prefrontal and entorhinal cortices, and cerebellar cortex. Relative to the low level of GR, mineralocorticoid receptor (MR) mRNA and protein expression were abundant within the DG and CA of the rhesus monkey hippocampal formation. These results indicate that, in the primate, neocortical and hypothalamic areas may be more important targets for GR-mediated effects of glucocorticoids than the hippocampus. Alternatively, it is also possible that glucocorticoid effects are mediated through the MRs present in the hippocampal formation. PMID- 10844036 TI - c-Fos expression in GABAergic, serotonergic, and other neurons of the pontomedullary reticular formation and raphe after paradoxical sleep deprivation and recovery. AB - The brainstem contains the neural systems that are necessary for the generation of the state of paradoxical sleep (PS) and accompanying muscle atonia. Important for its initiation are the pontomesencephalic cholinergic neurons that project into the pontomedullary reticular formation and that we have recently shown increase c-Fos expression as a reflection of neural activity in association with PS rebound after deprivation in rats (Maloney et al. , 1999). As a continuation, we examined in the present study c-Fos expression in the pontomedullary reticular and raphe neurons, including importantly GABAergic neurons [immunostained for glutamic acid decarboxylase (GAD)] and serotonergic neurons [immunostained for serotonin (Ser)]. Numbers of single-labeled c-Fos+ neurons were significantly increased with PS rebound only in the pars oralis of the pontine reticular nuclei (PnO), where numbers of GAD+/c-Fos+ neurons were conversely significantly decreased. c-Fos+ neurons were positively correlated with PS, whereas GAD+/c-Fos+ neurons were negatively correlated with PS, suggesting that disinhibition of reticular neurons in the PnO from locally projecting GABAergic neurons may be important in the generation of PS. In contrast, through the caudal pons and medulla, GAD+/c-Fos+ cells were increased with PS rebound, covaried positively with PS and negatively with the electromyogram (EMG). In the raphe pallidus obscurus, Ser+/c-Fos+ neurons were positively correlated, in a reciprocal manner to GAD+/c-Fos+ cells, with EMG, suggesting that disfacilitation by removal of a serotonergic influence and inhibition by imposition of a GABAergic influence within the lower brainstem and spinal cord may be important in the development of muscle atonia accompanying PS. PMID- 10844037 TI - Chronic hypersensitivity for inflammatory nociceptor sensitization mediated by the epsilon isozyme of protein kinase C. AB - We have identified a mechanism, mediated by the epsilon isozyme of protein kinase C (PKCepsilon) in peripheral neurons, which may have a role in chronic inflammatory pain. Acute inflammation, produced by carrageenan injection in the rat hindpaw, produced mechanical hyperalgesia that resolved by 72 hr. However, for up to 3 weeks after carrageenan, injection of the inflammatory mediators prostaglandin E(2) or 5-hydroxytryptamine or of an adenosine A(2) agonist into the same site induced a markedly prolonged hyperalgesia (>24 hr compared with 5 hr or less in control rats not pretreated with carrageenan). A nonselective inhibitor of several PKC isozymes and a selective PKCepsilon inhibitor antagonized this prolonged hyperalgesic response equally. Acute carrageenan hyperalgesia could be inhibited by PKA or PKG antagonists. However, these antagonists did not inhibit development of the hypersensitivity to inflammatory mediators. Our findings indicate that different second messenger pathways underlie acute and prolonged inflammatory pain. PMID- 10844038 TI - Long-term rAAV-mediated gene transfer of GDNF in the rat Parkinson's model: intrastriatal but not intranigral transduction promotes functional regeneration in the lesioned nigrostriatal system. AB - Previous studies have used recombinant adeno-associated viral (rAAV) vectors to deliver glial cell line-derived neurotrophic factor (GDNF) in the substantia nigra to protect the nigral dopamine (DA) neurons from 6-hydroxydopamine-induced damage. However, no regeneration or functional recovery was observed in these experiments. Here, we have used an rAAV-GDNF vector to express GDNF long-term (6 months) in either the nigral DA neurons themselves, in the striatal target cells, or in both of these structures. The results demonstrate that both nigral and striatal transduction provide significant protection of nigral DA neurons against the toxin-induced degeneration. However, only the rats receiving rAAV-GDNF in the striatum displayed behavioral recovery, accompanied by significant reinnervation of the lesioned striatum, which developed gradually over the first 4-5 months after the lesion. GDNF transgene expression was maintained at high levels throughout this period. These results provide evidence that rAAV is a highly efficient vector system for long-term expression of therapeutic proteins in the nigrostriatal system. PMID- 10844039 TI - Predicting vulnerability to acoustic injury with a noninvasive assay of olivocochlear reflex strength. AB - Permanent noise-induced damage to the inner ear is a major cause of hearing impairment, arising from exposures occurring during both work- and pleasure related activities. Vulnerability to noise-induced hearing loss is highly variable: some have tough, whereas others have tender ears. This report documents, in an animal model, the efficacy of a simple nontraumatic assay of normal ear function in predicting vulnerability to acoustic injury. The assay measures the strength of a sound-evoked neuronal feedback pathway to the inner ear, the olivocochlear efferents, by examining otoacoustic emissions created by the normal ear, which can be measured with a microphone in the external ear. Reflex strength was inversely correlated with the degree of hearing loss after subsequent noise exposure. These data suggest that one function of the olivocochlear efferent system is to protect the ear from acoustic injury. This assay, or a simple modification of it, could be applied to human populations to screen for individuals most at risk in noisy environments. PMID- 10844040 TI - Modeling LGN responses during free-viewing: a possible role of microscopic eye movements in the refinement of cortical orientation selectivity. AB - Neural activity appears to be essential for the normal development of the orientation-selective responses of cortical cells. It has been proposed that the correlated activity of LGN cells is a crucial component for shaping the receptive fields of cortical simple cells into adjacent, oriented subregions alternately receiving ON- and OFF-center excitatory geniculate inputs. After eye opening, the spatiotemporal structure of neural activity in the early stages of the visual pathway depends not only on the characteristics of the environment, but also on the way the environment is scanned. In this study, we use computational modeling to investigate how eye movements might affect the refinement of orientation tuning in the presence of a Hebbian scheme of synaptic plasticity. Visual input consisting of natural scenes scanned by varying types of eye movements was used to activate a spatiotemporal model of LGN cells. In the presence of different types of movement, significantly different patterns of activity were found in the LGN. Specific patterns of correlation required for the development of segregated cortical receptive field subregions were observed in the case of micromovements, but were not seen in the case of saccades or static presentation of natural visual input. These results suggest an important role for the eye movements occurring during fixation in the refinement of orientation selectivity. PMID- 10844041 TI - Chemical determinants of the rat electro-olfactogram. AB - The chemical properties that determine the distribution of the electro olfactogram were studied after exposure of a large area of the rat olfactory epithelium. Multiple electrodes were placed along the rostral border of endoturbinate IV on the midline of the nasal cavity. This array of electrodes spanned a region containing the four receptor gene expression zones described for the rat. The response to a series of odorants containing only carbon, hydrogen, and oxygen was strongly related to electrode position. For most hydrocarbons, the responses were progressively larger toward the ventral epithelium. The only exceptions were aromatic hydrocarbons, which evoked nearly equal response sizes across the epithelium. Ketones and aldehydes evoked relatively larger dorsal responses than did hydrocarbons with similar structures. Aromatic ketones and aldehydes evoked systematically larger responses from the dorsal part of the epithelium. The response profiles for most odorants were well described by a linear fit to the electrode position along the dorsal-ventral position on the epithelium. However, a few bicyclic odorants and carboxylic acids evoked significantly nonlinear profiles. It is concluded that there is a systematic distribution of odorant sensitivity across this part of the epithelium and that this sensitivity is related to general chemical properties. Other evidence suggests that these properties extend to other parts of the epithelium. This spatial sensitivity of the epithelium to odorants probably contributes to olfactory coding in parallel with the convergence of axons from olfactory sensory neurons expressing the same receptor type. PMID- 10844042 TI - Adult learning deficits after neonatal exposure to D-methamphetamine: selective effects on spatial navigation and memory. AB - The effects of neonatal d-methamphetamine (MA) treatment on cued and spatial learning and memory were investigated. MA was administered to neonatal rats on postnatal days 11-20. All groups received four subcutaneous injections per day. Group MA40-4 received 40 mg. kg(-1). d(-1) of MA in four divided doses (10 mg/kg per injection). Group MA40-2 received 40 mg. kg(-1). d(-1) of MA in two divided (20 mg/kg/injection) and saline for the other two injections per day. Controls received saline for four injections per day. As adults, both MA groups showed no differences in swimming ability in a straight swimming channel. The MA40-4 group showed no differences in cued learning, but was impaired in hidden platform learning in the Morris water maze on acquisition. They also showed reduced memory performance on probe trials. Similar trends were seen on reversal learning and reversal probe trials. Reduced platform-size learning trials caused spatial learning impairments to re-emerge in the MA40-4 group. The MA40-2 group showed no differences in straight channel swimming, but was slower at finding the visible platform during cued learning. They were also impaired during acquisition and memory trials in the Morris hidden platform maze. They showed a similar trend on reversal learning and memory trials, but were not different during reduced platform-size learning trials. When the MA40-2 group's performance on hidden platform learning and memory trials was adjusted for cued trial performance, the spatial learning deficits remained. Deficits of spatial learning and memory are a selective effect of neonatal methamphetamine treatment irrespective of other learning and performance variables. PMID- 10844043 TI - Social status controls somatostatin neuron size and growth. AB - Many animal species show flexible behavioral responses to environmental and social changes. Such responses typically require changes in the neural substrate responsible for particular behavioral states. We have shown previously in the African cichlid fish, Haplochromis burtoni, that changes in social status, including events such as losing or winning a territorial encounter, result in changes in somatic growth rate. Here we demonstrate for the first time that changes in social status cause changes in the size of neurons involved in the control of growth. Specifically, somatostatin-containing neurons in the hypothalamus of H. burtoni increase up to threefold in volume in dominant and socially descending animals compared with cell sizes in subordinate and socially ascending fish. Because somatostatin is known to be an inhibitor of growth hormone release, the differences in cell size suggest a possible mechanism to account for the more rapid growth rates of subordinate and socially ascending animals compared with those of dominant or socially descending fish. These results reveal possible mechanisms responsible for socially induced physiological plasticity that allow animals to shift resources from reproduction to growth or vice versa depending on the social context. PMID- 10844044 TI - Sustained visual attention performance-associated prefrontal neuronal activity: evidence for cholinergic modulation. AB - Cortical cholinergic inputs are hypothesized to mediate attentional functions. The present experiment was designed to determine the single unit activity of neurons within the medial prefrontal cortex (mPFC) of rats performing a sustained visual attention task. Demands on attentional performance were varied by the presentation of a visual distractor. The contribution of cholinergic afferents of the mPFC to performance-associated unit activity within this area was determined by recording neuronal activity before and after unilateral cholinergic deafferentation using intracortical infusion of the immunotoxin 192 IgG-saporin. Presentation of the visual distractor resulted in a decrease in the detection of brief, unpredictable visual signals. As predicted, the unilateral loss of cholinergic inputs within the recording area of the mPFC did not affect sustained attentional performance. Cholinergic deafferentation, however, resulted in a decrease in the overall firing rate of medial prefrontal neurons and a substantial reduction in the proportion of neurons whose firing patterns correlated with specific aspects of behavioral performance. Furthermore, cholinergic deafferentation attenuated the frequency and amplitude of increased mPFC neuronal firing rates that were associated with the presentation of the visual distractor. The main findings from this experiment suggest that cholinergic inputs to the mPFC strongly influence spontaneous and behaviorally correlated single unit activity and mediate increases in neuronal activity associated with enhanced demands for attentional processing, all of which may be fundamental aspects in the maintenance of attentional performance. PMID- 10844045 TI - Local disparity not perceived depth is signaled by binocular neurons in cortical area V1 of the Macaque. AB - Binocular neurons that are closely related to depth perception should respond selectively for stimuli eliciting an appropriate depth sensation. To separate perceived depth from local disparity within the receptive field, sinusoidal luminance gratings were presented within a circular aperture. The disparity of the aperture was coupled to that of the grating, thereby rendering unambiguous the psychophysical matching between repeating cycles of the grating. In cases in which the stimulus disparity differs by one horizontal period of the grating, the portion of the grating that locally covers a receptive field is binocularly identical, but the depth sensation is very different because of the aperture. For 117 disparity-selective V1 neurons tested in two monkeys, the overwhelming majority responded equally well to configurations that were locally identical but led to different perceptions of depth. Because the psychophysical sensation is not reflected in the firing rate of V1 neurons, the signals that make stereo matches explicit are most likely elaborated in extrastriate cortex. PMID- 10844046 TI - Corneal biopsy in the management of progressive microbial keratitis. AB - PURPOSE: To study the indications and role of diagnostic corneal biopsy in the management of patients with progressive microbial keratitis. METHODS: The records of 33 consecutive patients who underwent a diagnostic corneal biopsy from January 1, 1986, to December 31, 1998, were reviewed. The indication for corneal biopsy was progressive infectious keratitis despite intensive broad-spectrum topical antimicrobial therapy, or progressive stromal infiltration inaccessible to corneal scrapings. Microbiologic evaluation of all corneal biopsies was performed, and 11 of the 33 biopsies were also examined histopathologically. RESULTS: A microorganism was isolated from 27 (82%) of the 33 corneal biopsies. Of the six patients with a negative biopsy, a penetrating keratoplasty was performed in five patients and the pathogen was identified by examination of the corneal button. In one patient no microorganism was identified; however, the infection resolved with topical antimicrobial therapy. The most common risk factor for keratitis was foreign body exposure or corneal abrasion (14 patients). A solid stromal infiltrate was the most common pattern of corneal involvement. Corneal biopsy revealed previously unidentified microorganisms that led to a change in antimicrobial therapy in 24 (89%) of the 27 patients and confirmed prior culture results in the remaining 3 patients. Microbiologic evaluation of the corneal biopsy was more sensitive than histopathologic examination. Acanthamoeba was the most commonly isolated pathogen (five cases), followed by Propionibacterium acnes and Fusarium (four cases each). Gram-positive organisms were isolated in 16 patients. Only five of the 27 patients with a positive corneal biopsy required a penetrating keratoplasty, in contrast to five of the six patients with a negative corneal biopsy (P =.005). During the 13-year period of the study, only three corneal biopsies were performed in the last 6 years. CONCLUSIONS: Microbiologic evaluation of a diagnostic corneal biopsy contributed significantly to the diagnosis, treatment, and outcome of patients with progressive infectious keratitis. PMID- 10844047 TI - Posterior capsular opacification with hydrogel, polymethylmethacrylate, and silicone intraocular lenses: two-year results of a randomized prospective trial. AB - PURPOSE: To compare the visual outcome, percentage of posterior capsular opacification, and laser capsulotomy rates with polymethylmethacrylate, silicone, and hydrogel intraocular lens implants at 1 and 2 years postoperatively. METHODS: Ninety-three eyes of 93 patients were randomized to receive a polymethylmethacrylate, silicone, or hydrogel intraocular lens implant. A standardized surgical protocol was followed by a single surgeon using phacoemulsification with capsulorhexis; any patients with surgical complications were excluded, and all patients received standardized medication and follow-up. Patients were examined at days 1 and 7, months 1, 3, and 6, and years 1 and 2 after surgery. At each assessment, best-corrected logMAR visual acuity and Pelli Robson contrast sensitivity were measured. Posterior capsular opacification was objectively assessed by digital retroillumination imaging with the use of a dedicated software program and calculated as the percentage area of opacified capsule. Laser capsulotomy was performed if the eye had lost 2 lines of visual acuity with a clinically opaque capsule. RESULTS: At 2 years postoperatively, the mean percentage area of posterior capsular opacification for hydrogel lenses was 63%; for polymethylmethacrylate, 46%; and for silicone, 17%. Hydrogel intraocular lenses were associated with 17% more posterior capsule opacification than were polymethylmethacrylate lenses (95% confidence interval, 1-33; P =. 037) and 45% more than were silicone lenses (95% confidence interval, 33-58; P <.0001) at 2 years. Polymethylmethacrylate lenses had 28% more posterior capsule opacification than silicone lenses (95% confidence interval, 13-43; P <.0001) at 2 years. Twenty-eight percent of patients with hydrogel intraocular lenses required an Nd:YAG laser posterior capsulotomy at 2 years, compared with 14% with polymethylmethacrylate, whereas no patients with silicone lenses needed a capsulotomy (P =.014). Visual acuity was not significantly different among the three groups, but patients with silicone intraocular lenses had significantly better contrast sensitivity than those with hydrogel lenses (P =.046). CONCLUSIONS: Intraocular lenses made of this specific hydrogel were associated with a significantly higher degree of posterior capsular opacification and more laser capsulotomies than polymethylmethacrylate and silicone intraocular lenses. PMID- 10844048 TI - Cataract and phthisis bulbi after laser photoablation for threshold retinopathy of prematurity. AB - PURPOSE: To describe the visual and structural outcome of eyes that developed a dense cataract after laser photoablation for threshold retinopathy of prematurity. METHODS: A retrospective review of eight consecutive infants who developed dense cataract(s) after bilateral laser photoablation for threshold retinopathy of prematurity. Of the 10 eyes with cataract, five eyes were treated with a diode laser and five with an argon laser. The stage and zone of the retinopathy of prematurity, number of burns applied, time of onset of the cataract, clinical findings at the time of cataract surgery, and the course after cataract surgery were reviewed. RESULTS: Six eyes had zone 1 disease and four had zone 2 disease. The mean number of burns applied per eye was 2532 +/- 856 (range, 1400 to 4500). A cataract was diagnosed a median of 3 [corrected] weeks (range, 1 to 28 weeks) after laser photoablation. Nine of the 10 cataracts were sufficiently dense to preclude a view of the fundus. All 10 eyes had clinical signs suggestive of an inflammatory or ischemic process that included one or more of the following findings: corneal edema, pupillary membrane, iris atrophy, depigmentation of ciliary processes, pigment on the anterior lens surface, posterior synechiae, hyphema, and shallow anterior chamber. Nine eyes underwent cataract surgery. Five of the 10 eyes had retinal detachment ranging in severity from stage 4A to stage 5 at the time of cataract surgery. Nine of the 10 eyes progressed to phthisis bulbi and no light perception. CONCLUSIONS: A dense cataract developing in the eye of an infant after laser photoablation for threshold retinopathy of prematurity is associated with a poor visual prognosis. The constellation of associated clinical findings appears to be most consistent with anterior segment ischemia. PMID- 10844049 TI - Relationship between structural abnormalities and short-wavelength perimetric defects in eyes at risk of glaucoma. AB - PURPOSE: To determine the relative prevalence of blue-yellow perimetric defects and structural abnormalities of the optic nerve and nerve fiber layer in eyes at risk of glaucoma. METHODS: Seventy-two eyes (of 72 patients) at risk of glaucoma, with normal white-on-white full threshold perimetry, were examined prospectively with blue-yellow full-threshold perimetry (Humphrey). Structural evaluations were conducted with qualitative assessment of stereoscopic color optic disk photographs and monochromatic nerve fiber layer photographs performed independently by three masked examiners (a glaucoma specialist and two glaucoma fellows), and statistical analysis of summary parameters was obtained with scanning confocal laser tomography (abnormal defined as values outside 95% confidence limits established in normal control subjects). RESULTS: Kappa values for interobserver agreement were 0.64, 0.88, and 0.79 for optic disk evaluation and 0.59, 0.60, and 0.61 for nerve fiber layer evaluation. Thirteen (18%) of 72 eyes had blue-yellow abnormalities. A total of 30 eyes (42%) were identified as having a structural abnormality; 29 (40%) had qualitatively determined optic disk abnormalities, 21 (29%) had qualitatively determined nerve fiber layer defects, and 26 (36%) had statistically significant structural abnormalities. Twelve of 13 eyes with blue-yellow defects had a detectable structural abnormality; all 12 had abnormalities identified with disk photography, nine with nerve fiber layer photography, and 12 by scanning laser tomography. CONCLUSIONS: Clinically detectable structural abnormalities frequently coexist with blue-yellow perimetric defects in patients with ocular hypertension. A substantial proportion of patients with ocular hypertension with normal blue-yellow perimetry has early detectable glaucomatous structural abnormalities. PMID- 10844050 TI - Measurement of peripapillary retinal nerve fiber layer volume in glaucoma. AB - PURPOSE: To measure peripapillary retinal nerve fiber layer volume in normal subjects and patients with ocular hypertension, primary open-angle glaucoma, and low-tension glaucoma. METHODS: Sixty-five subjects were classified into four groups of normal subjects and subjects with ocular hypertension, primary open angle glaucoma, and low-tension glaucoma on the basis of intraocular pressure measurements, visual field loss, and optic disk appearance. Groups were matched for sex, age, and optic disk area. Peripapillary retinal nerve fiber layer volume measurements were made with a modification of software version 1.11 of the Heidelberg Retina Tomograph confocal scanning laser ophthalmoscope. Retinal nerve fiber layer volume measurements were taken at 0.1-mm increments from the disk margin for a global 360-degree assessment and at four predefined segments using two different reference planes. Statistical analysis was carried out using analysis of variance with Bonferroni correction. RESULTS: Retinal nerve fiber layer volume measurements showed a gradation from normal to ocular hypertension and from ocular hypertension to glaucoma groups. Mean group measurements showed statistically significant differences (P <.05) in peripapillary retinal nerve fiber layer volume for most segmental measures between the groups. Measurements with the default reference plane in the 0.0-mm to 0.1-mm and 0.1-mm to 0.2-mm increments for the superotemporal segment showed the greatest differences between groups (P <.00005). CONCLUSION: Peripapillary retinal nerve fiber layer volume measurements differ between groups of normal subjects and patients with ocular hypertension, primary open-angle glaucoma, and low-tension glaucoma. This measure offers a further method of assessment of retinal nerve fiber layer in patients with glaucoma and glaucoma suspects. PMID- 10844051 TI - Pathophysiology of and prophylaxis against late ahmed glaucoma valve occlusion. AB - PURPOSE: To understand the pathophysiology of late Ahmed Glaucoma Valve failures and devise strategies to minimize this problem. METHODS: One hundred sixty eyes that had undergone Ahmed Glaucoma Valve implants by one surgeon were retrospectively reviewed. Six eyes with late (greater than 3 months) Ahmed Glaucoma Valve occlusion requiring operative intervention were identified. Two of these eyes underwent initial successful transcameral drainage tube irrigation and four initially required Ahmed Glaucoma Valve exchange. Intraoperative images, postoperative histologic analysis, and Ahmed Glaucoma Valve handling experiments were performed. RESULTS: Two eyes with late occlusion (33%; 1.25% of total implants) were initially successfully treated with irrigation alone. A gap between the valve cover and valve body junction allowed fibrovascular ingrowth and produced valve failure ultimately in five of six eyes (83%; 3.1% of total implants). This gap could be produced by grasping the device along the center line, indenting the valve cover, and damaging the plastic rivets attaching the valve cover to the valve body. Handling the Ahmed Glaucoma Valve outside this "no touch zone" eliminated this problem. CONCLUSIONS: Leaflet adhesion has a low incidence and may be treated by transcameral drainage tube irrigation. Late onset distal occlusion is best treated by Ahmed Glaucoma Valve exchange with respect for the "no touch zone." Respecting the "no touch zone over the valve mechanism should avoid creation of gaps between the valve cover and valve body junction, which allow secondary fibrovascular ingrowth. PMID- 10844052 TI - Open-angle glaucoma associated with Graves disease. AB - PURPOSE: To estimate the prevalence of open-angle glaucoma, including normal tension glaucoma, in association with Graves disease. METHODS: In a prospective study, a complete ophthalmic examination including applanation tonometry, exophthalmometry, automated static threshold perimetry, and computed tomography of the orbit was performed in 104 consecutive Japanese patients with Graves disease (age range, 11 to 86 years; mean, 42 years; 80 women, 24 men). Ocular hypertension was defined as an intraocular pressure greater than 21 mm Hg on at least two occasions during the period of follow-up. RESULTS: Of the 104 patients, 14 (13%) exhibited typical glaucomatous visual field defects in automatic static threshold perimetry in the absence of compressive optic neuropathy. The intraocular pressure in seven of the 14 patients was consistently less than 21 mm Hg during the follow-up period. Thus, these patients were diagnosed as having normal-tension glaucoma. Of the 104 patients, 23 (22%) were diagnosed as having ocular hypertension. CONCLUSIONS: The prevalence of normal-tension glaucoma as well as open-angle glaucoma and ocular hypertension was significantly higher among patients with Graves disease than in the general population. PMID- 10844053 TI - Antioxidant agents in the treatment of Graves' ophthalmopathy. AB - PURPOSE: To report the effect of antioxidant agents in the treatment of mild and moderately severe Graves' ophthalmopathy. METHODS: Prospective, nonrandomized, comparative study performed at a referral center. A series of 11 patients with mild or moderately severe, active, newly diagnosed Graves' ophthalmopathy were included in the study. Allopurinol (300 mg daily) orally and nicotinamide (300 mg daily) orally were used for 3 months. A complete ophthalmologic examination was performed before and 1 and 3 months after initiation of treatment. The response to treatment was estimated separately for each component of the disease and overall by its effect on a total eye score. Eleven patients with mild or moderately severe, active, newly diagnosed Graves' ophthalmopathy who received placebo were also examined at the same time points. Patients in each group were recruited consecutively. Although nonsmoking was not an exclusion criterion, all patients were cigarette smokers. RESULTS: Nine (82%) of 11 patients treated with oral antioxidants showed improvement of mild to moderately severe Graves' ophthalmopathy versus three (27%) of 11 patients in the control group (P <.05). Soft tissue inflammation was the component of the disease that responded more to treatment. No side effects of antioxidant treatment were recorded. Patients' satisfaction was high. CONCLUSIONS: This pilot study presents encouraging results in the treatment of mild and moderately severe Graves' ophthalmopathy with antioxidant agents. To evaluate these preliminary results, randomized prospec tive studies are needed. PMID- 10844054 TI - Estrogen-replacement therapy: effects on retrobulbar hemodynamics. AB - PURPOSE: Estrogen-replacement therapy causes vasodilation and increased blood flow in major peripheral arteries. We examined the role that estrogen may play in enhancing perfusion within the watersheds of several major and minor retrobulbar arteries. METHODS: Postmenopausal women receiving estrogen-replacement therapy (n = 16) were compared with both age-matched women not receiving estrogen (n = 16) and with young women (n = 20). Studies involved color Doppler imaging analysis of flow velocities measured in the ophthalmic, central retinal, and nasal and temporal posterior ciliary arteries. RESULTS: In the ophthalmic artery, young women and postmenopausal women receiving estrogen exhibited reduced resistance indexes as compared with postmenopausal women not receiving estrogen (each P <. 001). In contrast, flow velocities in the central retinal artery were similar among the three groups of women. In the posterior ciliary arteries, a different pattern emerged: young women, as compared with either group of postmenopausal women, showed greater peak systolic and end-diastolic velocities at similar resistance index (each P <.05). CONCLUSIONS: Estrogen-replacement therapy in postmenopausal women apparently helps reduce vascular resistance distal to the ophthalmic artery to levels matching those of young women. However, estrogen replacement has little impact on flow velocities in the posterior ciliary arteries. In those vessels, aging per se may reduce perfusion, potentially contributing to the age-dependent risk of major eye diseases, such as glaucoma and age-related macular degeneration. PMID- 10844055 TI - Pseudoexfoliation syndrome in Australian adults. AB - PURPOSE: To describe the prevalence and correlates of pseudoexfoliation syndrome in Australians aged 40 years and older. METHODS: Cluster, stratified sampling was employed to identify a cohort representative of the population of the state of Victoria aged 40 years and older that included urban, rural, and nursing home residents. A standardized personal interview and clinical eye examination, including intraocular pressure, were performed at locally established test sites. The presence of any pseudoexfoliation material on the iris or lens capsule was noted on dilated slit-lamp examination. Participants were classified as having pseudoexfoliation syndrome if any pseudoexfoliation material was present in either eye. Univariate analyses with t tests and chi-square were first employed to evaluate risk factors for pseudoexfoliation. Any factors with P <.10 were then fitted in a backward stepwise logistic regression model. For the final multivariate models, P <.05 was considered statistically significant. RESULTS: A total of 3,271 of the urban residents (83% of eligible), 403 nursing home residents (90% of eligible), and 1473 rural residents (92%) participated. The urban residents ranged in age from 40 to 98 years (mean = 59), and 1,511 (46%) were men. The nursing home residents ranged in age from 46 to 101 years (mean 82), and 85 (21%) were men. The rural residents ranged in age from 40 to 103 years (mean 60), and 701 (47.5%) were men. Participants with bilateral cataract extraction were excluded from further analyses. The overall rate of pseudoexfoliation syndrome in this population was 0.98% (95% confidence limit = 0.57, 1.28). The prevalence of pseudoexfoliation material in either eye increased significantly with age. No cases of pseudoexfoliation syndrome were observed in people aged 90 years and older. However, people with bilateral cataract surgery had been excluded from these analyses. After adjusting for age and cataract, only glaucoma remained significantly related to pseudoexfoliation (odds ratio = 3.80, 95% confidence limit = 1.73, 8.33). DISCUSSION: In conclusion, we found only two strong correlates of pseudoexfoliation in our population-based sample of Victorians aged 40 years and older: age and glaucoma. PMID- 10844056 TI - Immune recovery uveitis in patients with AIDS and cytomegalovirus retinitis after highly active antiretroviral therapy. AB - PURPOSE: To estimate the incidence and describe the characteristics of immune recovery uveitis in patients with acquired immunodeficiency syndrome (AIDS) and cytomegalovirus retinitis treated with highly active antiretroviral therapy. METHODS: The records of all patients with AIDS and cytomegalovirus retinitis from 1995 to 1998 seen at the AIDS Ophthalmology Service of the Johns Hopkins Medical Institutions were reviewed. Eighty-two patients with cytomegalovirus retinitis treated with highly active antiretroviral therapy were identified. Thirty-three patients (40.2%) were classified as responders to highly active antiretroviral therapy, defined as an increase in CD4+ T-cell count by 50 cells/microL or more to a level of 100 cells/microL or more. RESULTS: Immune recovery uveitis occurred in six patients. Among the 33 patients with an immunologic response to highly active antiretroviral therapy, the incidence rate of immune recovery uveitis was 0.109/person-year. Ocular complications associated with immune recovery uveitis included cystoid macular edema (four patients), epiretinal membranes (two patients), and optic disk neovascularization (one patient). CONCLUSIONS: Immune recovery uveitis was uncommon in our population but may have vision-impairing complications. PMID- 10844057 TI - Hyperhomocysteinemia: a risk factor for central retinal vein occlusion. AB - PURPOSE: Previous studies have documented that elevated plasma homocysteine level is a risk factor for vascular disease. This study was performed to determine whether hyperhomocysteinemia is a risk factor for central retinal vein occlusion. METHODS: In a case-control study, data from 74 patients with documented central retinal vein occlusion were reassessed. Control subjects consisted of individuals referred to the same clinic for assessment of a nonretinal vascular disease. Hyperhomocysteinemia was defined as a total plasma homocysteine level above the 95th percentile in the control group. RESULTS: The mean total plasma homocysteine level was 11.58 +/- 4.67 micromol/l (range, 5-26 micromol/l) for cases, and 9.49 +/- 2.65 micromol/l (range, 5-20 micromol/l) for control subjects. Of the 74 patients with a central retinal vein occlusion, 16 (21.6%) had total plasma homocysteine levels above the 95th percentile in the control group (odds ratio, 6.53; 95% confidence interval, 1.81-23.50; P =. 003). Hyperhomocysteinemia was present in five (55%) of the nine individuals with bilateral disease, nine (30%) of the 30 patients with ischemic occlusions, and 45 (31%) of the 83 eyes with severe visual loss. CONCLUSION: Hyperhomocysteinemia is a risk factor for central retinal vein occlusion and may suggest a poor prognosis in patients with central retinal vein occlusion. PMID- 10844058 TI - Ocular trauma in the United States Army: hospitalization records from 1985 through 1994. AB - PURPOSE: To determine the incidence of hospitalized ocular injury in the United States Army and evaluate specific types and external causes of these injuries. METHODS: A US Army database that captured all hospital discharge records for Army personnel admitted to military and civilian hospitals was used to determine incident episodes of ocular injury requiring hospitalization from 1985 through 1994. Denominator data were available from the US Army. RESULTS: The average annual incidence of hospitalization for a principal or secondary diagnosis of ocular trauma (total hospitalized ocular injury) was 77.1 per 100,000 persons (95% confidence interval, 75.1-79.2). There was a 38% decline in the rate of total hospitalized ocular injury during this 10-year period. Men had twice the rates of women over all age groups. The highest rate occurred in the 17- to 19 year age group, with rates of 220.7 and 123.4 per 100,000 in men and women, respectively. Whites had a higher rate than blacks and nonwhites-nonblacks. Almost a third of the injuries were contusions of the eye and adnexa. Among men, the leading causes were machinery or tools (21%), fights (18%), transport accidents (18%), and sports and training (11%). Only 7% were related to weaponry or war, and of these, 90% were from nonbattle activities. CONCLUSION: The type and cause of injury suggest that preventive measures may be effective in decreasing the incidence of ocular trauma requiring hospitalization in US Army personnel. PMID- 10844059 TI - Necrobiotic xanthogranuloma: long-term outcome of ocular and systemic involvement. AB - PURPOSE: To report long-term outcomes of patients with necrobiotic xanthogranuloma, to investigate the propriety of therapeutic surgical excision or debulking, and to study tissue specimens by immunoperoxidase staining and in situ hybridization. METHODS: Medical records of all patients at the Mayo Clinic, Rochester, Minnesota, with necrobiotic xanthogranuloma between 1980 and 1997 were reviewed. A follow-up letter was sent to each patient inquiring about the current status of the lesions, the treatment regimen, and associated systemic diseases. RESULTS: The average age (+/- standard deviation) of the 15 men and 11 women was 56.8 +/- 14.8 years. Of the 26 patients, 21 (81%) had lesions of the ocular adnexa. Ulceration of the lesions occurred in 11 patients (42%). The lesions recurred after surgical removal in 11 patients (42%) and on prior incision sites from unrelated operations in three patients (12%). The average duration of follow up from the appearance of characteristic skin lesions was 10 +/- 6.1 years. Four patients had multiple myeloma, five had a plasma cell dyscrasia, and one had a lymphoproliferative disorder during this period. Time to development of associated malignancy ranged from 8 years before the skin lesions to 11 years after the skin lesions. Overall survival was 100% at 10 years and 90% at 15 years (95% confidence limit, 0.73 to 1.00). Immunoperoxidase stains demonstrated that most histiocytes are not of Langerhans cell lineage. Monoclonal immune globulins were not identified in tissue specimens. CONCLUSION: Care of patients with necrobiotic xanthogranuloma should include avoidance of surgical removal, if possible, and lifelong follow-up to detect the development of associated malignancy. PMID- 10844060 TI - Evidence of genetic heterogeneity in autosomal recessive congenital fibrosis of the extraocular muscles. AB - PURPOSE: Autosomal recessive congenital fibrosis of the extraocular muscles (CFEOM2) has been described in families from Saudi Arabia. Affected individuals have ptosis and exotropic ophthalmoplegia, and their disease has been mapped to chromosome 11q13. Here, we describe the phenotypic findings in a similarly affected Yemenite family and analyze the family for linkage to the CFEOM2 locus, as well as to the autosomal dominant CFEOM1 and CFEOM3 loci on chromosomes 12cen and 16q24, respectively. METHODS: The family was ascertained through two affected daughters. There are four unaffected siblings, and the parents are consanguineous. Each family member was examined, and linkage analysis was performed using markers from the CFEOM1, CFEOM2, and CFEOM3 loci. RESULTS: Both affected daughters have congenital bilateral ophthalmoplegia. The 15-month-old proband has restrictive exotropia. She fixates with either eye in abduction and with a compensatory head turn to the opposite side. Her 4-year-old sister has a small exotropia and severely limited eye movements. All other family members have normal ophthalmologic examinations. Genetic analysis excluded linkage of the family's disease to the CFEOM2 and CFEOM3 loci. A lod score of 2.0 (the maximum possible, given the family size and structure), was obtained at the CFEOM1 locus, and the alleles reduced to homozygosity in both affected daughters and none of the other children. CONCLUSIONS: These data establish that there is genetic heterogeneity in autosomal recessive CFEOM and suggest that this second recessive locus may be allelic to the autosomal dominant CFEOM1 locus at 12cen. PMID- 10844062 TI - Gelatino-lattice corneal dystrophy: clinical features and mutational analysis. AB - PURPOSE: To report five unrelated Japanese individuals with "gelatino-lattice corneal dystrophy that clinically resembled, to some extent, gelatinous drop-like corneal dystrophy and lattice corneal dystrophy type 1. METHODS: Genomic DNA isolated from the five individuals with "gelatino-lattice corneal dystrophy was used as a template for polymerase chain reaction to amplify all exons of the candidate gene betaig-h3 and M1S1. The polymerase chain reaction product was then sequenced. RESULTS: In all cases, betaig-h3 was mutated in "gelatino-lattice corneal dystrophy (Arg124Cys), which is the same nucleotide change examined previously in lattice corneal dystrophy type 1. On the other hand, no mutation was detected in the entire coding region of M1S1. CONCLUSION: Based on the results of this study, it is suggested that "gelatino-lattice corneal dystrophy may be a subtype of lattice corneal dystrophy type 1. PMID- 10844061 TI - Chronic clinical course of two patients with severe corneal dystrophy caused by homozygous R124H mutations in the betaig-h3 gene. AB - PURPOSE: To report the chronic clinical course of two patients with homozygous R124H mutations in the betaig-h3 gene. METHODS: Case reports. RESULTS: Two patients with homozygous R124H mutations in the betaig-h3 gene developed severe juvenile corneal opacities that required keratoplasty. After surgery, corneal opacities recurred and limited the recovery of visual acuity in the chronic follow-up. CONCLUSION: In patients with homozygous R124H mutations in the betaig h3 gene, recurrence of corneal opacities after keratoplasty limits the recovery of visual acuity in the chronic follow-up. PMID- 10844063 TI - Keratoconus and mitral valve prolapse. AB - PURPOSE: To investigate the prevalence of keratoconus in patients with mitral valve prolapse (MVP). METHODS: Videokeratography was performed in 36 patients (72 eyes) with an echocardiographic diagnosis of MVP, and were compared with 25 controls. All participants underwent slit-lamp examination and completed a medical questionnaire about joint hypermobility, arthralgia, asthma, and allergic reactions. RESULTS: Keratoconus was found in eight eyes (11. 1%) (P =.056) of eight patients (22.2%) (P =.049) in the MVP group and in one eye (2%) of one patient (4%) in the control group. The keratoconus was unilateral and asymptomatic in all cases. Conjunctival papillae were statistical significant higher in the MVP group (P =.015). A statistically significant intergroup difference existed in the presence of an allergic reaction (P =.001), but not in arthralgia, hypermobility of joints, or asthma. CONCLUSION: A borderline important association of MVP with keratoconus and with allergy exists. Patients with mitral valve prolapse complaining of poor vision should be suspected of having keratoconus. PMID- 10844064 TI - Visual field defect associated with laser in situ keratomileusis. AB - PURPOSE: To report a case of visual field defect associated with laser in situ keratomileusis. METHODS: Case report. A 28-year-old woman with high myopia (-10D) and a family history of normal tension glaucoma underwent bilateral laser in situ keratomileusis keratorefractive surgery. Preoperatively, both eyes had normal intraocular pressure and visual field. RESULTS: At the first postoperative visit 1 day after apparently uncomplicated laser in situ keratomileusis, the patient reported a scotoma in the right eye. At 3-month follow-up, visual fields revealed the patient had developed a near-superior altitudinal visual field defect in the right eye. The defect did not progress over 1 year of follow-up examinations. CONCLUSION: Increased intraocular pressure associated with the microkeratome vacuum ring used during laser in situ keratomileusis may have precipitated optic nerve head ischemia and visual field defect. PMID- 10844065 TI - Decreased substance P concentrations in tears from patients with corneal hypesthesia. AB - PURPOSE: To determine whether substance P concentrations in tears reflect corneal hypesthesia. METHODS: Sixteen patients with unilateral corneal hypesthesia participated in this study. Unstimulated tears were collected from each eye of all subjects. Substance P concentrations in tears were measured by an enzyme immunoassay system. RESULTS: Substance P concentrations in tears from the affected eye were lower than those in tears from the unaffected eye in all subjects. The mean concentration of substance P in tears from affected eyes was 197.7 +/- 69.5 pg/mL, markedly lower than those from unaffected eyes (333.2 +/- 74.6 pg/mL). CONCLUSION: Substance P concentrations in tears, which are thought to reflect the neuropeptides levels in ocular tissues, can be used as an indicator of corneal denervation. PMID- 10844066 TI - Herpes simplex virus DNA identification from aqueous fluid in Fuchs heterochromic iridocyclitis. AB - PURPOSE: To report the presence of herpes simplex virus DNA in the aqueous humor of an eye with Fuchs heterochromic iridocyclitis. METHODS: In an eye with a clinical diagnosis of Fuchs heterochromic iridocyclitis, samples of aqueous humor and anterior capsule of the lens were obtained during cataract surgery. Polymerase chain reaction was performed on the samples to detect the presence of viral DNA including herpes simplex virus, varicella-zoster virus, and cytomegalovirus. Serologic analysis was also performed for antiviral immunoglobulins. RESULTS: Herpes simplex virus DNA was identified in the aqueous humor but not in the anterior capsule. Serum immunoglobulin G was positive for herpes simplex virus, varicella-zoster virus, and cytomegalovirus. CONCLUSIONS: The presence of herpes simplex virus DNA in the aqueous humor of an eye with Fuchs heterochromic iridocyclitis suggests that herpes simplex virus infection may play a role in the pathogenesis of Fuchs heterochromic iridocyclitis. PMID- 10844067 TI - Intraocular lacrimal gland choristoma involving iris and ciliary body. AB - PURPOSE: To describe clinical and pathologic features of an iridociliary lacrimal gland choristoma in which the diagnosis was made clinically and confirmed histopathologically. METHODS: Case report with clinical, ultrasound biomicroscopic, and histopathologic observations. RESULTS: We examined a 12-month old male for a large iris mass that had been present since birth. We suspected it to be a lacrimal gland choristoma based on characteristic clinical features. The lesion was removed by iridocyclectomy, and the diagnosis of lacrimal gland choristoma was confirmed. CONCLUSION: Intraocular lacrimal gland choristoma has unique clinical features that should suggest the diagnosis. PMID- 10844068 TI - No clinically important effects on intraocular pressure after short-term administration of sildenafil citrate (Viagra). AB - PURPOSE: To assess the short-term effects of sildenafil citrate on intraocular pressure in healthy male volunteers and participants in clinical trials. METHODS: Intraocular pressure and pupil diameter were measured in two placebo-controlled studies. Oral doses of sildenafil citrate (VIAGRA; Pfizer Inc, New York, New York) ranged from 10 mg to 150 mg. RESULTS: No major changes in intraocular pressure or pupillometry were detected at any time (1.0-24 hours) after administration of sildenafil. Additionally, of 36 subjects with a medical history of increased intraocular pressure in the sildenafil safety database, none were reported to have a clinically significant increase of their intraocular pressure. During clinical trials, two glaucoma cases were listed as serious adverse events, but were not considered treatment related. CONCLUSION: No clinical abnormalities were observed in intraocular pressure or pupil diameter in subjects receiving sildenafil. Currently, no evidence suggests that long-term treatment with sildenafil has an effect on intraocular pressure or is associated with the development or worsening of glaucoma. PMID- 10844069 TI - Arcus senilis and mortality in a population with diabetes. AB - PURPOSE: To estimate the prevalence of arcus senilis and its association with mortality in a diabetic population. METHODS: A cohort of persons with younger (n = 996) and older onset (n = 1,370) diabetes was examined. Mortality information was obtained from death certificates. RESULTS: Prevalence of arcus senilis increased with age and was higher in men than in women. In the younger onset group, it was also associated with a history of cardiovascular disease. As a risk factor for mortality after controlling for age and sex, arcus senilis was not associated with death from all causes, ischemic heart disease, or stroke. CONCLUSION: Arcus senilis provides no more information about mortality risk than age of the person. PMID- 10844070 TI - Hepatocyte growth factor levels in the vitreous of patients with proliferative vitreoretinopathy. AB - PURPOSE: To determine whether the hepatocyte growth factor is increased in the vitreous of patients with proliferative vitreoretinopathy. METHODS: We assayed hepatocyte growth factor levels by enzyme-linked immunosorbent assay (ELISA) in vitreous samples from 65 consecutive eyes (65 patients) with proliferative vitreoretinopathy (23 eyes), rhegmatogenous retinal detachment (22 eyes), and macular hole or idiopathic epiretinal membrane (control subjects, 20 eyes) having undergone pars plana vitrectomy. RESULTS: Vitreous levels of hepatocyte growth factor were 3.94 +/- 2.29 (mean +/- SD) ng/ml in proliferative vitreoretinopathy, 2.02 +/- 0.84 ng/ml in rhegmatogenous retinal detachment, and 2.16 +/- 1.39 ng/ml in the control subjects. The vitreous levels in proliferative vitreoretinopathy were much greater than levels in rhegmatogenous retinal detachment (P =.0002) and in the control subjects (P =.0007). In proliferative vitreoretinopathy, there was a tendency toward higher levels in eyes with grade D by the Retina Society terminology. CONCLUSION: The results suggest the possibility that hepatocyte growth factor may play a role in the pathogenesis of proliferative vitreoretinopathy. PMID- 10844071 TI - Spontaneous central retinal artery occlusion in hemoglobin sickle cell disease. AB - PURPOSE: To describe a case of spontaneous central retinal artery occlusion in a young man with hemoglobin sickle cell disease. METHOD: Case report. RESULTS: A 31 year-old African-American man with a history of hemoglobin sickle cell disease developed sudden painless loss of vision in the right eye. Medical history was remarkable for the recent history of a mild painful crisis, but no other systemic illness or contributing factors. Central retinal artery occlusion was diagnosed with retinal whitening, cherry red spot, and delayed arteriovenous transit on fluorescein angiography. Over the ensuing week, the patient had visual recovery to 20/60 in the absence of therapeutic intervention. CONCLUSION: Central retinal artery occlusion has been reported in sickle cell hemoglobinopathies (ie, SS, S thal, sickle trait, and sickle cell), but the association with sickle cell disease is rare. Most reports have described additional contributing factors, such as trauma or concomitant systemic illness, to help account for the central retinal artery occlusion. The present case suggests that sickle cell disease alone is sufficient for the development of central retinal artery occlusion. PMID- 10844072 TI - Susac syndrome. AB - PURPOSE: To report the first Japanese case of the Susac syndrome characterized by microangiopathy of the brain, retina, and cochlea. METHODS: Case report. A 36 year-old Japanese man underwent extensive clinical and laboratory examination for encephalopathy, bilateral recurrent branch retinal arterial occlusion, and hearing loss. RESULTS: This case was initially diagnosed as acute disseminated encephalomyelitis. Despite exhaustive clinical and laboratory examination of the patient, no evidence of a known systemic disease was found. The bilateral recurrent branch retinal arterial occlusions were widely disseminated. Remission occurred 1 year after symptom onset. This patient demonstrated all of the clinical features associated with the Susac syndrome. CONCLUSION: Susac syndrome has been considered rare, but it is probably more common than previously thought and seems to occur in any race. This disorder usually occurs in young women, but it can affect men. PMID- 10844073 TI - Atypical reginal vasculitis associated with ticlopidine hydrochloridine use. AB - PURPOSE: To describe two cases of retinal vasculitis shortly after the initiation of ticlopidine hydrochloride (Ticlid, Roche, Kingsland St, NJ) therapy. METHODS: Case reports of two patients. The first patient was a 43-year-old white woman complaining of spots, floaters, and flashes of lights in both eyes 3 weeks after the initiation of treatment with ticlopidine hydrochloride. The second patient was a 72-year-old woman complaining of decreased visual acuity in the left eye for 2 weeks, 4 weeks after initiating oral administration of ticlopidine hydrochloride. RESULTS: Both patients had resolution of the vasculitis after the discontinuation of ticlopidine therapy. CONCLUSION: The temporal relation and the resolution of symptoms after discontinuation of ticlopidine hydrochloride suggest that the vasculitis was related to the ticlopidine hydrochloride administration. Knowledge of this potential complication of ticlopidine hydrochloride is important for the early diagnosis of this possible drug-induced side effect and the cessation of ticlopidine hydrochloride. PMID- 10844074 TI - Migration of silicone oil into the brain: a complication of intraocular silicone oil for retinal tamponade. AB - PURPOSE: To report a case in which intravitreal silicone oil migrated along the intracranial portion of the optic nerve and into the lateral ventricles of the brain after the repair of a retinal detachment secondary to cytomegalovirus retinitis. METHODS: A 42-year-old man with acquired immunodeficiency syndrome (AIDS) developed a rhegmatogenous retinal detachment in his left eye secondary to a cytomegalovirus infection of the retina. The detachment was repaired using 5000 cs intraocular silicone oil for a long-term tamponade. Subsequently, the affected eye developed glaucoma, which was poorly controlled. Fifteen months after the retinal surgery, he developed a peripheral neuropathy that was thought to be AIDS related. Computed tomography and magnetic resonance imaging of the head were performed to investigate the neuropathy. RESULTS: The patient was found to have a foreign substance within his lateral ventricles that shifted with position and was identical with respect to its imaging properties to the remaining intraocular silicone oil. Additional material was found along the intracranial portion of his optic nerve. CONCLUSION: Under certain circumstances, intraocular silicone oil may migrate out of the eye, along the intracranial portion of the optic nerve, and into the lateral ventricles of the brain. PMID- 10844075 TI - Intraocular hemorrhages associated with endoscopic spinal surgery. AB - PURPOSE: To report the occurrence of intraocular hemorrhages in association with endoscopic spinal surgery. METHODS: Case report. RESULTS: A 40-year-old patient noted severe visual loss in both eyes after epidural endoscopy, epidurography, and endoscopic adhesiolysis. Ophthalmic examination showed a best-corrected visual acuity of RE: 20/800, and LE: counting fingers, and extensive subhyaloid, retinal, and subretinal hemorrhages in both eyes. The hemorrhages resolved over a period of 8 weeks and the best-corrected visual acuity improved to 20/40 in both eyes after 4 months. CONCLUSION: A syndrome of visual loss and intraocular hemorrhages after an endoscopic spinal procedure is studied. PMID- 10844076 TI - Branch retinal artery occlusion after thyroid artery interventional embolization. AB - PURPOSE: To report a case of branch retinal artery occlusion after thyroid artery interventional embolization. METHODS: A 33-year-old man with hyperthyroidism complained of visual loss and scotoma in the left eye after thyroid artery interventional embolization. He underwent a full ophthalmologic examination, including fluorescein angiography. RESULTS: Visual acuity was 20/25, with inferior and superior scotomas present in the left eye. Fluorescein angiography of the left eye revealed delayed filling of a superotemporal branch retinal artery and nonfilling of an inferotemporal branch retinal artery. CONCLUSION: A small, but definite risk of retinal artery occlusion after thyroid artery interventional embolization should be considered. PMID- 10844077 TI - Tarsal fibroma. AB - PURPOSE: To report a case of an isolated tarsal fibroma. METHODS: Case report. Excisional biopsy and histopathological evaluation were performed on a solid lesion originating from the tarsal conjunctival surface of an upper eyelid. RESULTS: Histopathological evaluation, including positive trichrome stains, was consistent with fibroma of the left upper tarsus. No recurrence has developed after a follow-up interval of a year. CONCLUSION: Tarsal fibroma is a rare condition that should be considered in the differential diagnosis of tarsal lesions. PMID- 10844078 TI - Congenital orbital varices causing extreme neonatal proptosis. AB - PURPOSE: To report a 6-week-old male with extreme proptosis caused by thrombosed orbital varices. METHODS: A 6-week-old male presented with advanced proptosis of his left eye. Computed tomography and magnetic resonance imaging studies of the orbits revealed a heterogeneous cystic mass that filled the entire left orbit. Extreme proptosis and corneal exposure prompted urgent surgical excision of the mass. Histopathologic review of the lesion was consistent with orbital varices. CONCLUSIONS: Although orbital varices usually are found in adults, they should be considered in the differential diagnosis of orbital lesions in children. Observation is usually warranted, but surgical intervention may be necessary in advanced cases. PMID- 10844079 TI - Treatment of optic nerve sheath meningioma with three-dimensional conformal radiation. AB - PURPOSE: To report a patient recovering vision after three-dimensional conformal radiotherapy for optic nerve sheath meningioma. METHODS: Radiotherapy was delivered by a three-dimensional conformal technique in 28-180-cGy fractions. RESULTS: Visual acuity improved from 20/200 to 20/30, and the visual field defect resolved. CONCLUSION: Visual loss from optic nerve sheath meningioma can be reversed by three-dimensional conformal radiotherapy. PMID- 10844080 TI - Researchers and regulators reflect on first gene therapy death PMID- 10844081 TI - The value of routine preoperative medical testing before cataract surgery PMID- 10844082 TI - Cost-effectiveness of public-funded options for cataract surgery in mysore, india PMID- 10844083 TI - Air bags and ocular injuries PMID- 10844084 TI - Acute hydrops in the corneal ectasias: associated factors and outcomes PMID- 10844085 TI - Indeterminate melanocytic proliferations of the conjunctiva PMID- 10844086 TI - Giant papillary conjunctivitis in frequent-replacement contact lens wearers: a retrospective study PMID- 10844088 TI - Two clinical trials of an intraocular steroid delivery system for cataract surgery PMID- 10844087 TI - Keratocyte density in vivo after photorefractive keratectomy in humans PMID- 10844089 TI - Epilepsy surgery, visual fields, and driving: a study of the visual field criteria for driving in patients after temporal lobe epilepsy surgery with a comparison of goldmann and esterman perimetry PMID- 10844091 TI - Retinopathy and nephropathy in patients with type 1 diabetes four years after a trial of intensive therapy PMID- 10844090 TI - Localizing value of epileptic visual auras PMID- 10844092 TI - Non-steroidal anti-inflammatory drugs and apoptosis in the gastrointestinal tract: potential role of the pentose phosphate pathways. AB - Non-steroidal anti-inflammatory drugs (NSAIDs) are among the most widely prescribed drugs, primarily for treatment of arthritis. NSAIDs can have two effects independent of their anti-inflammatory action. In the stomach and small bowel long term NSAID consumption can lead to ulceration, whereas in the colon NSAID use can regress existing tumours. In this review, we hypothesise that NSAID induced damage occurs predominantly by promoting apoptosis, involving a number of mechanisms depending on the type and the redox state of the cell. In addition to inhibiting cyclooxygenase (COX) activity, this includes interfering with glucose metabolism through both arms of the pentose phosphate pathways and energy production via glycolysis and oxidative phosphorylation. Shifting the cellular balance from proliferation to apoptosis is probably the most important outcome by which NSAIDs exhibit their differing actions. Understanding how these different pathways can be reconciled and their contribution to the balance between cell birth and cell death is the challenge for the future. The pentose phosphate pathways may provide a pivotal point for understanding links between factors which alter proliferative activity (e.g. COXs), provide energy metabolism (particularly aerobic and anaerobic metabolism of glucose), and change the redox state of the cell leading to apoptosis. PMID- 10844093 TI - Inhibition of mechanotransducer currents in crayfish sensory neuron by CGS 9343B, a calmodulin antagonist. AB - The effects of CGS 9343B (zaldaride maleate), a calmodulin antagonist, on mechanosensitive channels were examined in crayfish slowly adapting sensory neurons using the two-electrode voltage clamp technique. In addition to its inhibition of voltage-gated Na(+) and K(+) currents, CGS 9343B (<30 microM) blocked reversibly the receptor current in a dose-dependent and voltage-dependent manner with a dissociation constant (K(d)) of 26.8 microM. The time course of the block was 265 s. Within the extension range of 3-30%, the reduction in receptor current was stimulus-independent and the gating mechanisms were not affected. Extracellular Ca(2+) was not necessary for its blocking effects. No changes in passive muscle tension were observed in the presence of 20 microM CGS 9343B. These results suggest that CGS 9343B, as a calmodulin antagonist, can also block mechanosensitive channels, possibly by being incorporated into the lipid membrane and/or interacting with the channel protein. PMID- 10844094 TI - Influence of salmeterol and benzalkonium chloride on G-protein-mediated exocytotic responses of rat peritoneal mast cells. AB - The long-acting beta(2)-adrenoceptor agonist salmeterol and the invert soap benzalkonium chloride share physicochemically important structures, namely a polar head group and a long aliphatic chain. Low concentrations of benzalkonium chloride have been shown to inhibit exocytotic responses in rat peritoneal mast cells by selectively interacting with heterotrimeric G-proteins of the G(i)-type. The present study investigates whether salmeterol inhibits, independently of beta adrenoceptors, exocytotic responses of rat peritoneal mast cells induced by the direct agonists at G-proteins mastoparan or guanosine 5'-O-(3-thiotriphosphate) (++GTP gamma S++). Exocytosis was studied by secretion assays ([3H]5 hydroxytryptamine ([3H]5-HT)-release) using intact, streptolysin O-permeabilised or metabolically inhibited (antimycin, deoxyglucose) rat peritoneal mast cells. Both amphiphilics, salmeterol, and benzalkonium chloride, dose-dependently exerted biphasic effects on mastoparan-induced [3H]5-HT release in intact mast cells. In contrast to benzalkonium chloride, the dose-response curves for secretostatic and celltoxic effects of salmeterol markedly overlapped. Similar to benzalkonium chloride, salmeterol in non-cytotoxic concentrations (1-25 microg/ml) dose-dependently inhibited exocytosis induced by mastoparan (intact cells) or ++GTP gamma S (permeabilised cells). These findings indicate a direct, adrenoceptor-independent affection of G proteins by salmeterol in mast cells. PMID- 10844095 TI - Kinetic characterization of the nitric oxide toxicity for PC12 cells: effect of half-life time of NO release. AB - We investigated the effects of low concentrations of nitric oxide (NO) on cell viability using NO donors, (+/-)-(E)-4-methyl-2-[(E)-hydroxyimino]-5-nitro-6 methoxy-3-hex enamid e (NOR1), (+/-)-(E)-4-methyl-2-[(E)-hydroxyimino]-5-nitro-3 hexenamide (NOR2), (+/-)-(E)-4-ethyl-2-[(E)-hydroxyimino]-5-nitro-3-hexenamide (NOR3) and (+/-)-N-[(E)-4-ethyl-2-[(Z)-hydroxyimino]-5-nitro-3-hexen-1- yl]-3-pyr idine (NOR4). The half-life times of the NO release from these four NOR analogs, NOR1, NOR2, NOR3 and NOR4, were determined (6.5, 84, 105 and 340 min, respectively) by using 4,5-diaminofluorescein (DAF-2), a newly developed indicator of NO. Exposure of undifferentiated PC12 cells to low concentrations of NO donors, NOR2 or NOR3 (1-100 microM), but not NOR1 nor NOR4, resulted in cell death in a dose- and time-dependent manner, as determined from cell viability assessed by 3-(4,5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium (MTT) assay. After 24 h exposure to 50 microM NOR2 or NOR3, more than 90% of PC12 cells had died. Furthermore, while the toxic effect of NOR3 was attenuated by replacing the medium at 20 min, 1 or 2 h after drug addition, it was continued by replacing the medium at 3 h or later after drug addition. The cell death was characterized by DNA degradation, nuclear condensation and fragmentation, suggesting apoptosis like cell death. Pretreatment with an antioxidant ascorbic acid (0.1-0.5 mM) completely prevented the cell death caused by NOR3, while glutathione (0.1-0.2 mM) and cysteine (0.2-0.4 mM) provided partial protection. These findings suggest that the cell toxicity induced by NO at low concentrations strongly depends upon the duration of expose to NO from NO donors, and these toxic effects are effectively prevented by the antioxidant, ascorbic acid. PMID- 10844096 TI - Effect of dipyridamole and aspirin on the platelet-neutrophil interaction via the nitric oxide pathway. AB - This study was designed to determine the influence of the combination of aspirin and dipyridamole on the interaction in vitro between neutrophils and platelets through the nitric oxide (NO) pathway. Collagen-induced platelet aggregation (impedance method) was determined in platelet-rich plasma and in platelet-rich plasma+neutrophils, and cGMP (enzyme immunoanassay) and NO levels (electrochemical method, with a ISO-200 electrode) were also measured. The 50% inhibitory concentration (IC(50)) of aspirin was 139+/-11 microM in platelet-rich plasma, 367+/-21 microM in platelet-rich plasma+L-N(G)-nitro-arginine-methyl ester (L-NAME), and 42+/-3 microM in platelet-rich plasma+L-arginine. The IC(50) for dipyridamole in platelet-rich plasma was not affected by L-NAME or L arginine; the combination of aspirin with 20 microM dipyridamole (which has no effect per se) led to an IC(50) of 51+/-2 microM in platelet-rich plasma, 101+/-7 microM in platelet-rich plasma+L-NAME, and 13+/-2 microM in platelet-rich plasma+L-arginine. The cGMP levels showed the greatest increases in the aspirin plus dipyridamole group. Dipyridamole and aspirin increased the leukocyte production of NO: 50% increases were obtained at concentrations of 285+/-31 microM aspirin, 110+/-9 microM dipyridamole, and 16+/-2 microM aspirin+dipyridamole. Dipyridamole alone at a concentration of 20 microM had no significant effect on NO levels. We conclude that the combination of aspirin and dipyridamole significantly increases the antiplatelet effect of leukocytes, through an increase of NO, and that this effect is further evidence of the therapeutic benefits of this combination of drugs. PMID- 10844097 TI - Differential modulation of the GYKI 53784-induced inhibition of AMPA currents by various AMPA-positive modulators in cerebellar Purkinje cells. AB - The effects of various (S)-alpha-amino-3-hydroxy-5-methyl-4-izoxazole-propionate (AMPA) receptor modulators on AMPA-induced whole-cell currents were compared in isolated rat cerebellar Purkinje cells. The positive modulators, aniracetam, cyclothiazide, 1-(1, 3-benzodioxol-5-ylcarbonyl)-piperidine (1-BCP), and 1 (quinoxaline-6-ylcarbonyl)-piperidine (BDP-12), dose-dependently potentiated the steady-state component of AMPA currents. The negative modulator, (-)1-(4 aminophenyl)-4-methyl-7, 8-methylenedioxy-4,5-dihydro-3-methylcarbamoyl-2,3 benzodiazepine (GYKI 53784), dose-dependently suppressed AMPA responses. Its concentration-response curve was shifted to the right in a parallel fashion by all positive modulators, indicating a competitive type of interaction. However, the relative potencies of the positive modulators were different with regard to the enhancement of AMPA responses and the reversal of GYKI 53784-induced inhibition, respectively. It is supposed that positive modulators act at multiple allosteric sites and that they interact with GYKI 53784 at only one of these sites. PMID- 10844098 TI - Histamine H3 receptor-mediated suppression of inhibitory synaptic transmission in the submucous plexus of guinea-pig small intestine. AB - Conventional intracellular microelectrodes and marker injection techniques were used to study the actions of histamine on inhibitory synaptic transmission in the submucous plexus of guinea-pig small intestine. Bath application of histamine (1 300 microM) reversibly suppressed both noradrenergic and non-adrenergic slow inhibitory postsynaptic potentials in a concentration-dependent manner. These effects of histamine were mimicked by the selective histamine H(3) receptor agonist R(-)-alpha-methylhistamine but not the selective histamine H(1) receptor agonist, 6-[2-(4-imidazolyl)ethylamino]-N-(4-trifluoromethylphenyl) heptanecarboxamide (HTMT dimaleate), or the selective histamine H(2) receptor agonist, dimaprit. The histamine H(3) receptor antagonist, thioperamide, blocked the effects of histamine. Histamine H(1) and H(2) receptor antagonists did not change the action of histamine. Hyperpolarizing responses to focal application of norepinephrine or somatostatin by pressure ejection from micropipettes were unaffected by histamine and R(-)-alpha-methylhistamine. The results suggest that histamine acts at presynaptic histamine H(3) receptors on the terminals of sympathetic postganglionic fibers and intrinsic somatostatinergic nerves in the small intestine to suppress the release of the inhibitory neurotransmitters, norepinephrine and somatostatin. PMID- 10844099 TI - Facilitation and inhibition by endothelin-1 of adrenal catecholamine secretion in anesthetized dogs. AB - We examined the participation of endothelin ET(A) and ET(B) receptors in modulation by endothelin-1 of adrenal catecholamine secretion during cholinergic activation in pentobarbital-anesthetized dogs. Drugs were infused intra arterially into the adrenal gland. Splanchnic nerve stimulation (1 and 3 Hz) increased adrenal catecholamine output in a frequency-dependent manner. Endothelin-1 (0.2, 0.6, and 2 ng/kg/min) enhanced the catecholamine response induced by the 3-Hz nerve stimulation. Under pretreatment with an endothelin ET(A) receptor antagonist (R)-2-[(R)-2-[(S)-2-[[1-(hexahydro-1H azepinyl)]carbonyl]amino-4-m eth ylpentanoyl]amino-3-(2-pyridyl) propionic acid (FR139317) (1 microg/kg/min), endothelin-1 suppressed the 1- and 3- Hz nerve stimulation-induced catecholamine response in a dose-dependent manner. No inhibitory or facilitatory effect of endothelin-1 was observed under simultaneous pretreatment with FR139317 and an endothelin ET(B) receptor antagonist N-cis 2, 6 dimethylpiperidinocarbonyl-L-gamma-methylleucyl-D-1-met hox ycarbonyl tryptophanyl-D-norleucine (BQ-788) (1 microg/kg/min) or under pretreatment with BQ-788 alone. These results suggest that in the dog adrenal gland, endothelin-1 facilitates and inhibits adrenal catecholamine secretion during cholinergic activation by stimulating endothelin ET(A) and ET(B) receptors, respectively. PMID- 10844100 TI - Augmentation with a 5-HT(1A), but not a 5-HT(1B) receptor antagonist critically depends on the dose of citalopram. AB - Pharmacokinetic and pharmacodynamic parameters of the selective serotonin reuptake inhibitor 1-(3-dimethylaminopropyl)-1-(4-fluorophenyl)-5 phtalancarbonitril (citalopram) were determined in order to find optimal conditions for augmentation of its effect on extracellular serotonin [5 hydroxytryptamine (5-HT)] through blockade of 5-HT(1A) and 5-HT(1B) autoreceptors. Citalopram dose-dependently (0.3-10 micromol/kg s.c.) increased serotonin levels in ventral hippocampus of conscious rats. At plasma levels above approximately 0.15 microM, the effect of citalopram on extracellular 5-HT was augmented by both a 5-HT(1A) [N-[2-[4-(2-mehoxyphenyl)-1-piperazinyl]ethyl]-N-(2 pyridil) cyclohexa necarboxamide trihydrochloride (Way 100635), 1 micromol/kg s.c.] and a 5-HT(1B) receptor antagonist (2'-methyl-4'-(5-methyl-[1,2, 4]oxadiazol-3-yl)biphenyl-4-carboxylic acid [4-methoxy]-3-(4-methylpiperazin-1 yl)phenyl]amide (GR 127935), 1 micromol/kg s.c.). However, at plasma levels of the selective serotonin reuptake inhibitor below 0.15 microM, the effects of the antagonists diverged viz. the 5-HT(1B) receptor antagonist was still able to potentiate citalopram's effect on extracellular 5-HT, while the 5-HT(1A) receptor antagonist was no longer effective. These results suggest that in contrast to 5 HT(1B) autoreceptors, indirect activation of 5-HT(1A) autoreceptors by citalopram is critically related to the dose of selective serotonin reuptake inhibitor administered. The latter may have consequences for selective serotonin reuptake inhibitor augmentation strategies with 5-HT(1A) receptor antagonists in the therapy of depression and anxiety disorders. PMID- 10844101 TI - Neuroprotective effect of gamma-hydroxybutyrate in transient global cerebral ischemia in the rat. AB - The effect of gamma-hydroxybutyrate on the histological and behavioral consequences of transient brain ischemia was studied in the four vessel occlusion rat model. In saline-treated animals, 30 min ischemia caused a massive loss of neurons in the hippocampal CA1 subfield (normal neurons: 14%, 5%, 23% and 30% on the 3rd, 10th, 15th and 65th day after ischemia, respectively). gamma Hydroxybutyrate - 300 mg/kg intraperitoneally (i.p.) 30 min before or 10 min after arteries occlusion, followed by 100 mg/kg i.p. twice daily for the following 10 days - afforded a highly significant protection (normal neurons on the 3rd, 10th, 15th and 65th day after ischemia: 88% and 91%, 80% and 80%, 91% and 90%, 72% and 71% in rats receiving the first dose before or after arteries occlusion, respectively). The ischemia-induced sensory-motor impairment was significantly attenuated in rats receiving the first dose of gamma hydroxybutyrate before arteries occlusion. Finally, the ischemia-induced impairment in spatial learning and memory, evaluated starting 27 days after the ischemic episode, was significantly attenuated by gamma-hydroxybutyrate, either injected first at 30 min before or 10 min after arteries occlusion. Lower doses of gamma-hydroxybutyrate had no significant effect. In conclusion, these results indicate that gamma-hydroxybutyrate provides significant protection against both histological and behavioral consequences of transient global cerebral ischemia in rats. PMID- 10844102 TI - Diltiazem enhances the analgesic but not the respiratory depressant effects of morphine in rhesus monkeys. AB - There is evidence that blockade of Ca(2+) channels can modify the analgesia and respiratory depression produced by opioid drugs. The interaction between Ca(2+) channel blockade and drug-induced analgesia and respiratory depression was examined by administration of the L-type Ca(2+) channel blocker diltiazem together with various analgesic drugs. The antinociceptive effects of the drugs were evaluated using a warm-water (50 degrees C) tail-withdrawal assay in rhesus monkeys, and the respiratory depressant effects were evaluated using a pressure displacement plethysmograph. Pretreatment with diltiazem (10-40 mg/kg, i.m.) 30 min before administration of morphine (0.3 to 10 mg/kg) or heroin (0.03 to 1.0 mg/kg) produced a dose-dependent potentiation of the opioid-induced analgesia. The analgesic potency of morphine and heroin was increased by approximately 0.5 log unit in the presence of 40 mg/kg diltiazem. However, diltiazem failed to alter the analgesic potencies of the mu-opioid receptor agonists, fentanyl, etonitazene, nalbuphine, the kappa-opioid receptor agonist, U-50,488 [(trans)-3, 4-dichloro-N-methyl-N-[2-(1-pyrrolidinyl)-cyclohexyl]benzeneacetamide ], or the non-opioid, clonidine. Respiratory frequency, minute volume, and tidal volume were suppressed by morphine, heroin, and fentanyl, but these effects were not modified by pretreatment with diltiazem (40 mg/kg). These results suggest that diltiazem selectively potentiates morphine- and heroin-induced analgesia without modifying the effects of these opioids on respiration. PMID- 10844103 TI - Enantioselective behavioral effects of sibutramine metabolites. AB - The anti-obesity agent, racemic (RS)-sibutramine, has two active metabolites, desmethylsibutramine and didesmethylsibutramine. To the extent that sibutramine itself mediates some of its side effects, desmethylsibutramine and/or didesmethylsibutramine might be safer and just as therapeutically effective. Because both desmethylsibutramine and didesmethylsibutramine are also optically active, the present study assessed the anorexic effects (2.5-10 mg/kg, i.p., for all drugs), in rats, of the R(+)-and S(-)-enantiomers of both metabolites and compared them to the effects of racemic sibutramine. Locomotor activity (2.5-10 mg/kg, i. p., for all drugs), a dopamine dependent behavior, was also measured in view of some uncertainty regarding dopaminergic effects of sibutramine. In view of sibutramine's antidepressant profile in animal models, the same drugs were also tested in the Porsolt swim test (0.1-2.5 mg/kg, i.p., for all drugs). Lastly, the IC(50)s of all drugs to inhibit uptake in vitro of norepinephrine, serotonin and dopamine were determined. Both (R)-enantiomers had significantly greater anorexic effects than those of their respective (S)-enantiomers as well as of sibutramine. All of the agents increased locomotor activity and reduced immobilized time ("behavioral despair") in the swim test; again, the (R) enantiomers were more potent than the (S)-enantiomers and sibutramine. However, the anorexic and locomotor effects could be dissociated from each other as well as from effects in the swim test. Both (R)-desmethylsibutramine and (R) didesmethylsibutramine as well as sibutramine decreased food intake at a time (24 42 h post-treatment) when locomotor activity was unaffected. All of the drugs appeared to be more potent in the swim test than in the other tests and all of the drugs were more potent at inhibiting uptake of norepinephrine and dopamine than of serotonin. The results suggest that these enantioselective metabolites of sibutramine could be safe and effective treatments for obesity as well as possibly for depression. PMID- 10844104 TI - Nicotine induces disinhibitory behavior in the rat after subchronic peripheral nicotinic acetylcholine receptor blockade. AB - The present study investigated the effects of subchronic nicotine, mecamylamine and hexamethonium, alone or in combinations, on locomotor activity and behavioral inhibition. Rats were divided into groups and tested for locomotor activity after acute nicotine. The different groups received vehicle, nicotine, mecamylamine, mecamylamine+nicotine, hexamethonium (two different concentrations) and hexamethonium+nicotine injections once a day for 15 days after which they were tested for nicotine-induced locomotor activity again. Acutely, nicotine stimulated locomotor activity, and repeated daily nicotine or hexamethonium+nicotine administration sensitized the animals to this nicotine induced locomotor stimulation (locomotor sensitization). Mecamylamine administered subchronically in combination with nicotine was able to block the induction to locomotor sensitization to nicotine. None of the nicotinic receptor antagonists induced locomotor sensitization to nicotine by themselves. In the elevated plus-maze, subchronic nicotine treatment demonstrated a nicotine-induced behavioral disinhibition, measured as an increase of time spent in and entries made into open arms. In contrast to the findings regarding locomotor sensitization, none of the antagonists counteracted the induction of this nicotine-induced behavioral disinhibition after subchronic co-treatment with nicotine. In addition, both antagonists by themselves produced a similar effect as subchronic nicotine, i.e. promoted the development of nicotine-induced disinhibitory behavior. It was concluded that the induction of locomotor sensitization to nicotine involves stimulation of central nicotinic acetylcholine receptors, whereas the development of nicotine-induced behavioral disinhibition involves blockade of peripheral nicotinic acetylcholine receptors, and that the latter, but not the former, phenomenon from a pharmacological point of view appears to be related to the increased ethanol consummatory behavior observed after subchronic nicotine administration. PMID- 10844105 TI - Chronic administration of moxonidine suppresses sympathetic activation in a rat heart failure model. AB - Excessive sympathetic activity contributes to cardiovascular abnormalities, which negatively affect the prognosis of heart failure. The present study evaluated the effects of moxonidine, an imidazoline I(1) receptor agonist, on sympathetic activation and myocardial remodelling in a rat heart failure model. Rats were subjected to coronary artery ligation, and treated with moxonidine, 3 or 6 mg/kg/day, from 1 to 21 days after myocardial infarction. After 21 days, heart rate and blood pressure were measured in conscious, chronically instrumented rats. Plasma catecholamine levels were determined by high-performance liquid chromatography. Effects on post-myocardial infarction remodelling were evaluated from the ventricular weight body weight ratio and interstitial collagen deposition, measured morphometrically in the interventricular septum remote from the infarcted area. Moxonidine dose-dependently decreased myocardial infarction induced tachycardia but did not affect myocardial infarction reduced blood pressure. Plasma noradrenaline levels, which were elevated after myocardial infarction, decreased below sham-values with 6 mg/kg/day moxonidine. Ventricular weight-body weight ratio as well as interstitial collagen were significantly elevated in myocardial infarcted rats, and restored to sham values with 6 mg/kg/day moxonidine. These data suggest that moxonidine suppresses myocardial infarction induced sympathetic activation in a dose-dependent way as indicated by reduced heart rate and plasma noradrenaline levels. Furthermore, post-myocardial infarction remodelling may be attenuated at a higher dose-range of moxonidine as shown by normalisation of ventricular weight body weight ratio and interstitial collagen. PMID- 10844106 TI - Gender differences in the cardiovascular responses to morphine and naloxone in spinal rats. AB - Putative gender differences in opiate cardiovascular effects were evaluated in spinal rats. After a 4-h exposure to a single dose of morphine (30 mg/kg, i.v.), abstinence was precipitated by naloxone (0.03-3 mg/kg, i.v.). Morphine produced a long-lasting bradycardia and a transient increase in arterial pressure that was similar in both genders. Thereafter, blood pressure decreased both in males and females. Naloxone precipitated a similar dose-dependent heart rate increase in both sexes and a gender-dependent increase in blood pressure. This sex difference appeared in the shape of the response. Prazosin (0.2 mg/kg), prior to naloxone, reduced the pressor response in all animals, suggesting a similar participation of the noradrenergic system in both genders. The present results extend to acute dependence the notion of a sex-dependent differential effect of morphine. The need to consider gender as a factor when studying the effects of opioids is highlighted. PMID- 10844107 TI - Age-related involvement of the endothelium in beta-adrenoceptor-mediated relaxation of rat aorta. AB - The signalling pathway involved in beta-adrenoceptor relaxation was studied in aortas from rats either 8 or 54 weeks of age. The vasorelaxation produced by isoprenaline was almost completely abolished by endothelium removal in 54-week aortas, whereas in 8-week aortas, the effect was much smaller. The nitric oxide synthase inhibitor N-methyl-1-arginine acetate (L-NMMA) partially attenuated the isoprenaline induced relaxation to a similar extent in both age groups when the endothelium was intact, suggesting that although nitric oxide was involved, it could not explain the age-related difference. The K(+) channel inhibitor, tetraethylammonium inhibited isoprenaline vasorelaxation to a larger degree in 54 week compared to 8-week aortas indicating that K(+) channels were responsible for the age-related differences. We suggest that as the animals age, the smooth muscle cyclic AMP signalling system declines, and that this is compensated for by an alternate pathway involving the opening of K(+) channels. PMID- 10844108 TI - Post-junctional mechanisms involved in the potentiation of cardiac adrenergic responses by cocaine. AB - Cocaine cardiotoxicity is partly due to sympathetic activation of the heart resulting from inhibition of catecholamine uptake at the sympathetic nerve terminal and possible central sympathetic stimulation and/or inhibition. This study evaluated the role of postsynaptic mechanisms in potentiation by cocaine of cardiac adrenergic responses. Cardiovascular responses (arterial and left ventricular pressure, contractility and heart rate) to increasing doses of noradrenaline and to isoproterenol were obtained in anesthetized cats during a control period, after irreversible alpha-adrenoceptor blockade with phenoxybenzamine (5 mg/kg i.v.), and after cocaine (5 mg/kg, i.v.). Responses to noradrenaline were significantly reduced by phenoxybenzamine with lowering of the maximal rise of all parameters. Cocaine shifted the dose-response curve of noradrenaline to the left and enhanced its maximal effects. Some responses to isoproterenol, which is not taken up by nerve terminals, were also enhanced by cocaine. Pretreatment with chlorisondamine or verapamil prevented the cocaine induced enhancement of the maximal response to noradrenaline and the response to isoproterenol, but it did not inhibit potentiation of submaximal doses. Lidocaine did not potentiate the response to noradrenaline or isoproterenol. Use of chlorisondamine instead of cocaine potentiated responses to all noradrenaline doses and enhanced the responses to isoproterenol. These results suggest that the potentiation by cocaine of cardiac responses to adrenergic stimuli involves presynaptic mechanisms to block noradrenaline re-uptake, and postsynaptic mechanisms to raise the maximal responses. The latter may result from inhibition of central sympathetic outflow or from activation of cardiac Ca(+) channels, leading to increased cardiac sensitivity to noradrenaline. PMID- 10844109 TI - The antihypertensive and cardioprotective effects of (-)-MJ-451, an ATP-sensitive K(+) channel opener. AB - ATP-sensitive K(+) (K(ATP)) channel openers have been shown to be a potential class of therapeutic agents for the control of cardiovascular diseases, including angina, arrhythmias, and hypertension. In this study, the pharmacological activity of 6-cyano-3S,4R-dihydro-2, 2-dimethyl-2H-3-hydroxy-4-[5S-(1 hydroxymethyl)-2-oxo-1-pyrrolidinyl] -1-benzopyran ((-)-MJ-451), a synthetic K(ATP) opener, was evaluated in anesthetized rat models and in isolated rat thoracic rings. Results demonstrated that intravascular injection of (-)-MJ-451 (0. 02, 0.05 and 0.1 mg/kg) produced an immediate, dose-related reduction in mean arterial blood pressure in anesthetized spontaneously hypertensive rats (SHR), which persisted for more than 3 h and was not accompanied by reflex tachycardia. The hemodynamic changes were completely abolished by pretreatment with glibenclamide (4 mg/kg, i.v. bolus), a selective K(ATP) channel blocker. In isolated thoracic aorta, (-)-MJ-451 (10 nM-3 microM) produced a concentration dependent vasodilator effect on the phenylephrine (0.3 microM)-induced vasoconstriction. Moreover, (-)-MJ-451 relaxed the thoracic aorta contracted by low (5, 20 and 30 mM), but not high (40 and 60 mM) concentrations of extracellular potassium. In addition, (-)-MJ-451 showed cardioprotective effects in the rat model of 45-min left coronary artery occlusion followed by 1-h reperfusion. In myocardial ischemia, pretreatment with (-)-MJ-451 (2, 5 and 10 microg/kg, i.v. bolus) significantly reduced the incidence of ventricular fibrillation and the mortality, also reducing the total number of ventricular premature contractions, total duration of ventricular tachycardia and ventricular fibrillation. A significant reduction in infarct size was noted in three (-)-MJ 451 (2, 5 and 10 microg/kg)-treated groups. Also, the cardioprotective effects of (-)-MJ-451 were virtually abolished by pretreating the rats with glibenclamide (4 mg/kg, i.v. bolus). In conclusion, (-)-MJ-451, through opening the K(ATP) channel, exerted antihypertensive and cardioprotective effects. Therefore, it is suggested that (-)-MJ-451 has potential in the treatment of hypertension or acute myocardial infarction. PMID- 10844110 TI - Human internal mammary artery contraction by isoprostaglandin f(2alpha) type-III [8-iso-prostaglandin F(2alpha)]. AB - Isoprostaglandin F(2alpha) type-III (formerly known as 8-iso-prostaglandin F(2alpha)) is produced in large quantities in vivo in clinical situations associated with oxidant stress such as atherosclerosis, hypercholesterolemia, and myocardial reperfusion. Isoprostaglandin F(2alpha) type-III may alter smooth muscle and platelet functions. The aim of this study was to evaluate the effects of isoprostaglandin F(2alpha) type-III on isolated human internal mammary arteries, and to characterise the signalling underlying mechanisms. In organ baths, concentration-dependent contractions of human internal mammary arteries were obtained in response to isoprostaglandin F(2alpha) type-III stimulation. The responses to isoprostaglandin F(2alpha) type-III were inhibited in a concentration-dependent manner by the thromboxane A(2) receptor antagonist, GR 32191 ([1R-[1 alpha(Z), 2beta,3beta,5 alpha(+)-7-[[1, 1'-biphenyl)-4-yl]methoxy] 3-hydroxy-2-(1-piperidinyl) cyclo pentyl]-4-4heptanoic acid], hydrochloride), 3x10(-9) to 3x10(-7) M). However, this effect was associated with a decreased maximal contraction. AH 6809 (6-isopropoxy-9-oxoxanthene-2-carboxylic acid, 10( 6) to 3x10(-5) M), an EP(1)-DP receptor antagonist had no effect on isoprostaglandin F(2alpha) type-III-induced contractions. The maximal responses to isoprostaglandin F(2alpha) type-III were significantly reduced in the presence of the cyclooxygenase inhibitor indomethacin (10(-5) M) (E(max): 147+/-20% vs. 213+/-19% in control group, P<0.05). Isoprostaglandin F(2alpha) type-III stimulated thromboxane B(2) release (5.7-fold increase) from human internal mammary arteries. Baicaleine, a non-specific lipoxygenase inhibitor, (10(-4) M) and AA 861 (2,3,5-trimethyl-6-(12-hydroxy-5, 10-dodecadiynyl)-1,4 benzoquinone), a 5-lipoxygenase inhibitor (10(-5) M) did not affect isoprostaglandin F(2alpha) type-III response. In conclusion, this study shows that (1) isoprostaglandin F(2alpha) type-III is a vasoconstrictor in human internal mammary arteries, with a potency equivalent to prostaglandin F(2alpha), (2) the contractions induced by isoprostaglandin F(2alpha) type-III are mediated by TP receptor but not EP(1)-DP receptor activation, (3) thromboxane A(2) but not cysteinyl leukotrienes production is involved in the vascular effects of isoprostaglandin F(2alpha) type III. Isoprostaglandin F(2alpha) type-III, produced at sites of free radical generation, may play an important role in internal mammary artery spasm in situations of oxidant stress such as coronary bypass surgery. PMID- 10844111 TI - K(ATP) channels mediate the beta(2)-adrenoceptor agonist-induced relaxation of rat detrusor muscle. AB - We propose that ATP-sensitive K(+) (K(ATP)) channels are normally inactive but involved in beta(2)-adrenoceptor stimulated relaxation of the rat bladder. Spontaneous detrusor muscle contractions were unaffected by glibenclamide (K(ATP) channel blocker) but were reduced when pinacidil (K(ATP) channel opener) concentrations exceeded 10(-5) M. Inhibition by beta(2)-adrenoceptor agonist clenbuterol [10(-6) M] of 1 Hz electrical field stimulated contractions was abolished by glibenclamide [10(-6) M]. Glibenclamide [10(-6) M] decreased forskolin-induced relaxation [10(-9)-10(-4) M] in bladder muscle stimulated with 1 Hz electrical field. In the presence glibenclamide (10(-6) M) or myristoylated protein kinase A inhibitor (2)x[10(-6) M], clenbuterol [10(-9)-10(-5) M] failed to inhibit bladder contraction in response to 1 Hz electrical field stimulation. Therefore, K(ATP) channel opening and the subsequent hyperpolarization of cell membranes in response to beta(2)-adrenoceptor activation is mediated by raised cyclic-AMP levels and activation of protein kinase A. This counteracts ATP stimulated depolarization in bladder muscle, thereby reducing cell contraction. PMID- 10844112 TI - Prevention of pulmonary thromboembolism by NCX 4016, a nitric oxide-releasing aspirin. AB - We studied the antithrombotic activity of 2-acetoxybenzoate 2-[1-nitroxy-methyl] phenyl ester (NCX 4016), a novel nitric oxide (NO)-releasing aspirin derivative, in vivo in different animal models of platelet-dependent and independent pulmonary thromboembolism and compared it with that of aspirin. NCX 4016 protected mice from death induced by the intravenous (i.v.) injection of collagen plus epinephrine, of 9,11-dideoxy-11alpha, 9alpha-epoxymethano-prostaglandin F(2alpha) (U46619) and of thrombin while aspirin was only active against collagen plus epinephrine. The drop in platelet count and number of lung emboli were reduced by NCX 4016 more effectively than aspirin. NCX 4016 protected mice also from mechanical pulmonary embolism (i.v. injection of hardened rat red blood cells) while aspirin was ineffective. In rabbits, NCX 4016 significantly reduced the accumulation of [111In]oxine-labeled platelets in the pulmonary vasculature induced by collagen and by thrombin while aspirin produced reductions which were significant only versus collagen. In conclusion, NCX 4016 exerts a more pronounced antithrombotic activity than aspirin in vivo in two different animal species, largely due to a deeper inhibitory effect on platelets. NCX 4016 may represent a better antithrombotic agent than aspirin. PMID- 10844113 TI - Effects of inhaled glaucine on pulmonary responses to antigen in sensitized guinea pigs. AB - The alkaloid (S)-(+)-1,2,9,10-tetramethoxyaporphine (glaucine) is a phosphodiesterase 4 inhibitor with bronchodilator and anti-inflammatory activity in vitro. In this study, we examined the in vivo effects of glaucine on an animal model of asthma. In ovalbumin sensitized guinea pigs, inhaled glaucine (10 mg ml( 1), 3 min) inhibited the acute bronchoconstriction produced by aerosol antigen (antigen response was 256+/-42 and 95+/-14 cm H(2)O l(-1) s(-1) in control and glaucine-treated animals, respectively; P<0.05). Pretreatment with glaucine (10 mg ml(-1), 10 min inhalation, 30 min pre- and 3 h post-antigen exposure) markedly reduced airway hyperreactivity to histamine, eosinophil lung accumulation, and increased eosinophil peroxidase activity in bronchoalveolar lavage fluid 24 h after exposure of conscious guinea pigs to aerosol antigen. In addition, inhaled glaucine (5-10 mg ml(-1), 3 min) inhibited the microvascular leakage produced after inhaled antigen at all airway levels. These data support the potential interest of phosphodiesterase 4 inhibitors in asthma treatment. PMID- 10844114 TI - A novel adenosine analog, AMP579, inhibits neutrophil activation, adherence and neutrophil-mediated injury to coronary vascular endothelium. AB - We hypothesized that 1S-[1a,2b,3b, 4a(S*)]-4-[7-[[1-[(3-chloro-2 thienyl)methylpropyl]propyl-amino]-3H-i midazo[4,5-b] pyridyl-3-yl]-N-ethyl-2,3 dihydroxycyclopentane carboxamide (AMP579), a new adenosine analog, inhibits superoxide anion (O(2)(-)) generation and degranulation from canine neutrophils, neutrophil adherence and neutrophil-induced dysfunction to canine coronary artery endothelium by adenosine receptor-mediated mechanisms. AMP579 inhibited O(2)(-) generation (nM/20x10(6) neutrophils) from platelet activating factor (PAF) activated neutrophil in concentration-dependent manner (4.1+/-0.8 at 10 microM vs. 16.7+/-2.1 in PAF group, P<0.05). This inhibitory effect was blocked by the adenosine A(2A) receptor-selective antagonist, 4-(2-[7-Amino-2-(2 furyl)[1,2,4]triazolo[2,3-a][1,3, 5]triazin-5-ylamino]ethyl)phenol (ZM241385, 17.7+/-2.8, P<0.05), but not by either the adenosine A(1) receptor-selective antagonist, 8-Cyclopentyl-1.3-dipropylxanthine (DPCPX) or the adenosine A(3) receptor-selective antagonist, 9-Chloro-2-(2-furanyl)-5 [(phenylacetyl)amino][1,2,4]-triazolo[1, 5-c]quinazoline (MRS1220). AMP579 inhibited neutrophil degranulation dose-dependently by 38+/-2% at 10 microM (P<0.05). The inhibitory effect of AMP579 was not altered by either DPCPX or MRS1220, but was partially reversed by ZM241385 (69+/-8%, P<0.05 vs. AMP579 10 microM). A total of 10 microM AMP579 reduced neutrophil adherence to thrombin stimulated endothelium (neutrophils/mm(2)) from 269+/-16 to 44+/-4 (P<0.05); this was reversed by ZM241385, but not by DPCPX or MRS1220. After coincubation of unstimulated neutrophil with thrombin-stimulated endothelium, concentration relaxation responses to the endothelium receptor-dependent vasodilator, acetylcholine, were reduced (maximum 57+/-5% vs. 120+/-5% in controls, P<0.05). This endothelial dysfunction was attenuated by AMP579 (116+/-7%, P<0. 05). We conclude that AMP579 inhibits neutrophil activation and neutrophil-mediated coronary endothelial dysfunction, primarily by an adenosine A(2A) receptor mechanism. PMID- 10844115 TI - An anti-inflammatory ditriazine inhibiting leukocyte functions and expression of inducible nitric oxide synthase and cyclo-oxygenase-2. AB - A ditriazine derivative (4,10-dichloropyrido[5,6:4,5]thieno[3,2-d':3, 2-d]-1,2,3 ditriazine (DTD)) inhibited neutrophil functions, including degranulation, superoxide generation, and leukotriene B(4) production, without any effect on 5 lipoxygenase activity. This compound reduced nitric oxide (NO) and prostaglandin E(2) production in mouse peritoneal macrophages stimulated with lipopolysaccharide, whereas no influence on the activity of inducible NO synthase, cyclo-oxygenase-2 or cyclo-oxygenase-1 was observed. DTD significantly reduced mouse paw oedema induced by carrageenan and also markedly reduced NO and prostaglandin E(2) levels in exudates from 24-h zymosan-stimulated mouse air pouch. Western blot analysis showed that DTD reduced the expression of inducible NO synthase and cyclo-oxygenase-2. Our results indicate that DTD exerts anti inflammatory effects related to the inhibition of neutrophil functions and of NO and prostaglandin E(2) production, which could be due to a decreased expression of inducible NO synthase and cyclo-oxygenase-2. PMID- 10844116 TI - 1,5-Anhydro-D-fructose increases glucose tolerance by increasing glucagon-like peptide-1 and insulin in mice. AB - Besides being degraded to glucose-6-phosphate and to free glucose, glycogen is degraded by alpha-1,4-glucan lyase to 1, 5-anhydro-D-fructose. We examined the influence of 1, 5-anhydro-D-fructose on glucose-stimulated insulin secretion in vivo and in vitro in mice. When administered together with i.v. glucose (1 g/kg), 1,5-anhydro-D-fructose did not affect (at 0.2 g/kg) or inhibited (at 1 g/kg) insulin secretion without affecting glucose elimination. When incubated with isolated islets, 1, 5-anhydro-D-fructose at <16.7 mmol/l, did not affect glucose (11.1 mM)-stimulated insulin secretion but inhibited insulin secretion at 16.7 mmol/l. When given through a gastric gavage (150 mg/mouse) together with glucose (150 mg/mouse), 1,5-anhydro-D-fructose increased glucose tolerance and insulin secretion. Furthermore, 1, 5-anhydro-D-fructose potentiated the increase in plasma levels of the gut hormone, glucagon-like peptide-1 (GLP-1). We therefore conclude that when given enterally, but not parenterally, 1, 5-anhydro-D-fructose increases glucose tolerance in mice by increasing insulin secretion due to increased plasma levels of GLP-1. The sugar may therefore be explored for increasing endogenous GLP-1 secretion in the treatment of type 2 diabetes. PMID- 10844118 TI - Lack of effect of LY314582 (a group 2 metabotropic glutamate receptor agonist) on phencyclidine-induced locomotor activity in metabotropic glutamate receptor 2 knockout mice. AB - In metabotropic glutamate receptor 2 (mGlu(2)) knockout mice, the group 2 metabotropic glutamate receptor agonist LY314582 (20 mg/kg, i.p.), a racemate of LY354740, inhibits neither spontaneous nor phencyclidine (PCP)-induced (2.5 mg/kg, s.c.) locomotor activity. Since LY314582 attenuated spontaneous and PCP induced locomotor activity in wild-type control mice, these data indicate that the effects of LY314582 are mediated via the mGlu(2) receptor and not via the mGlu(3) receptor. PMID- 10844119 TI - Characterization of pancreastatin receptor and signaling in rat HTC hepatoma cells. AB - Pancreastatin, a chromogranin A-derived peptide widely distributed throughout the neuroendocrine system, has a general inhibitory effect on endocrine secretion and a counterregulatory effect on insulin action. We have recently described the cross-talk of pancreastatin with insulin signaling in rat hepatoma cells (HTC), where it inhibits insulin action and signaling through the serine phosphorylation of the insulin receptor, thereby impairing tyrosine kinase activity. Here, we have characterized pancreastatin receptors and signaling in HTC cells. The pancreastatin effector systems were studied by determining phospholipase C activity in HTC membranes and mitogen-activated protein kinase (MAPK) phosphorylation activity in HTC cells. Binding studies with radiolabeled pancreastatin showed a population of high affinity binding sites, with a B(max) of 8 fmol/mg protein and a K(d) of 0.6 nM. Moreover, we assessed the coupling of the receptor with a G protein system by inhibiting the binding with guanine nucleotide and by measuring the GTP binding to HTC membranes. We found that pancreastatin receptor was coupled with a G alpha(q/11) protein which activates phospholipase C-beta(1) and phospholipase C-beta(3), in addition to MAPK via both beta gamma and alpha(q/11). PMID- 10844120 TI - Regulation of bradykinin receptor gene expression in human lung fibroblasts. AB - In WI-38 human fibroblasts, interleukin-1 beta and tumour necrosis factor-alpha (TNF-alpha) increased bradykinin B(1) receptor mRNA, which peaked between 2 and 4 h, remaining elevated for 20 h. Binding of the bradykinin B(1) receptor selective ligand [3H]des-Arg(10)-kallidin, also increased, peaking at 4 h and remaining elevated for 20 h. The B(max) value for [3H]des-Arg(10)-kallidin rose from 280+/ 102 fmol/mg (n=3) to 701+/-147 fmol/mg (n=3), but the K(D) value remained unaltered (control, 1.04+/-0.33 nM (n=3); interleukin-1 beta, 0.88+/-0.41 nM (n=3)). The interleukin-1 beta-induced [3H]des-Arg(10)-kallidin binding sites were functional receptors, as bradykinin B(1) receptor agonist-induced responses increased in treated cells. Bradykinin B(2) receptor mRNA and [3H]bradykinin binding were upregulated by interleukin-1 beta, but not TNF-alpha. The effect of interleukin-1 beta on bradykinin B(2) receptors was smaller than for bradykinin B(1) receptors. Cycloheximide prevented interleukin-1 beta-mediated increases in B(1) and B(2) binding, but not mRNA suggesting that de novo synthesis of a transcriptional activator was unnecessary. PMID- 10844121 TI - The role of Arg(78) in the metabotropic glutamate receptor mGlu(1) for agonist binding and selectivity. AB - The metabotropic glutamate receptors belong to family C of the G-protein coupled receptor superfamily. These receptors all possess large extracellular amino terminal domains, where agonist binding takes place. We have previously constructed a molecular model of the amino terminal domain of the mGlu(1) receptor based on a weak amino acid sequence similarity with a family of bacterial periplasmic binding proteins (PBPs). The residues Ser(165) and Thr(188) were demonstrated to be involved in agonist binding to the receptor. Here, we report that mutation of Arg(78) in the mGlu(1b) receptor to leucine or glutamate completely knocks out [3H]quisqualic acid binding to the receptor. The constructed mutants, R78L and R78E, have also been characterized in a inositol phosphate assay. Here, the potency of (S)-glutamic acid and (S)-quisqualic acid was reduced 1000- and 100-fold, respectively, on R78L compared to the wild type (WT) receptor. (S)-Quisqualic acid was as potent on mutant R78E as it was on R78L, whereas (S)-glutamic acid was unable to activate R78E significantly at concentrations up to 10 mM. In conclusion, Arg(78) appears to be essential for agonist binding to the mGlu(1) receptor, most likely, through the formation of an ionic bond between its positively charged side chain and the distal acid group of the agonists. Furthermore, the different impact of the two mutations on (S) glutamic acid and (S)-quisqualic acid potencies strongly indicates that while Arg(78) appears to be a common site of interaction for the agonists, the Group I subtype selectivity of (S)-quisqualic acid is probably determined by other residues in the amino terminal domain. PMID- 10844122 TI - Selective opioid receptor agonist and antagonist displacement of [3H]naloxone binding in amphibian brain. AB - Opioid receptor ligands have been shown to elicit antinociception in mammals through three distinct types of receptors designated as mu, delta and kappa. These opioid receptors have been characterized and cloned in several mammalian species. Radioligand binding techniques were employed to characterize the sites of opioid action in the amphibian, Rana pipiens. Naloxone is a general opioid receptor antagonist which has not been characterized in R. pipiens. Kinetic analyses of [3H]naloxone in the amphibian yielded a K(D) of 6.84 nM while the experimentally derived K(D) value from saturation experiments was found to be 7.11 nM. Density data were also determined from saturation analyses which yielded a B(max) of 2170 fmol/mg. Additionally, K(i) values were calculated in competition studies for various unlabelled mu-, delta- and kappa-opioid receptor ligands to isolate their site of action. Highly selective antagonists for mu-, delta- and kappa-opioid receptors yielded nearly identical K(i) values against [3H]naloxone. PMID- 10844123 TI - Modulation of [3H]MK-801 binding to NMDA receptors in vivo and in vitro. AB - [3H]MK-801 binding in vivo was used to determine the occupancy of NMDA receptor ligands shown to allosterically modulate binding in vitro. ED(50) values (mg/kg) were obtained for the channel blockers (+)-5-methyl-10,11-dihydro-5,4 dibenzo[a,d]cyclohepten-5,10-imine maleate ((+)-MK-801, 0.2), 1-(1 phenylcyclohexyl)piperidine (phencyclidine, PCP, 1.7) and ketamine (4.4). Antagonists at the glutamate (DL-(2-carboxypiperazine-4-yl)propyl-1-phosphonate (DL-CPP, 5.7)) and glycine site (7-Chloro-4-hydroxy-3-(3-phenoxy)-phenyl 2(H)quinolinone (L-701,324, 14.1), 3R(+)cis-4-methyl-pyrrollid-2-one (L-687,414, 15.1)) inhibited [3H]MK-801 binding in vivo to varying maximum levels (69%, 103% and 45%, respectively). NR2B subunit-selective compounds acting at the ifenprodil site inhibited [3H]MK-801 in vivo by a maximum of 52-72% and gave ED(50) values (mg/kg) of: (+/-)-(1S*, 2S*)-1-(4-hydroxyphenyl)-2-(4-hydroxy-4-phenylpiperidino) 1-propanol ((+/-)CP-101,606), 1.9; (+/-)-(3R, 4S)-3-[4-(4-fluorophenyl)-4 hydroxypiperidin-1-yl]chroman-4,7-diol ((+/-)CP-283,097), 1.8; (+/-)-(R*, S*) alpha-(4-hydroxyphenyl)-beta-methyl-4-(phenylmethyl)-1-piperidine propanol ((+/ )Ro 25-6981), 1.0; ifenprodil, 6.0. The glycine site agonist D-serine stimulated binding to 151% of control with an ED(50) of 1.7 mg/kg. Results show that [3H]MK 801 binding in vivo may be used to measure receptor occupancy of ligands acting not only within the ion channel but also at modulatory sites on the NMDA receptor complex. PMID- 10844124 TI - Ecto-nucleotidase of cultured rat superior cervical ganglia: dipyridamole is a novel inhibitor. AB - Based on studies of agonist potencies on intact rat superior cervical ganglia, it has been suggested that this ganglion possesses distinct receptors for purine and pyrimidine nucleotides. However, the potency of an agonist is dependent upon whether it is susceptible to extracellular metabolism by the tissue. The aim of this investigation was to study the metabolism of uridine or adenosine nucleotides and nucleosides and the effects of dipyridamole and an ecto-ATPase inhibitor ARL 67156 (6-N, N-diethyl-D-beta-gamma-dibromomethylene-ATP) on their metabolism. Adenosine- and uridine-5'-triphosphates (ATP and UTP) were catabolised by cultured rat superior cervical ganglia, to their di- and monophosphates. Both ATP and UTP breakdown was significantly inhibited by dipyridamole (10 mcM), whereas ARL 67156 (100 mcM), was a weaker inhibitor of ATP degradation and inhibited UTP breakdown by approximately 40%. Metabolism of ATP and UTP by cultured rat superior cervical ganglia was reduced after treatment with cytosine-beta-arabinoside, suggesting that non-neuronal cells along with neuronal cells contribute to their breakdown. In conclusion, these results indicate that rat superior cervical ganglia possess ecto-nucleotidases capable of catabolising purine and pyrimidine nucleotides to their nucleosides, and that dipyridamole is a potent inhibitor of ecto-nucleotidase activity. PMID- 10844125 TI - Hyperpolarisation of rat mesenteric endothelial cells by ATP-sensitive K(+) channel openers. AB - Membrane potential responses of rat mesenteric endothelial cells were investigated in intact arteries using sharp electrodes. Levcromakalim, an activator of ATP-sensitive K(+) channels (K(ATP)) induced concentration-dependent hyperpolarisation of the endothelial cells, which was reversible by glibenclamide and ciclazindol, inhibitors of K(ATP). Another K(ATP) activator, diazoxide, also hyperpolarised the endothelial cells. Carbachol induced endothelial hyperpolarisation that was inhibited by combinations of apamin and charybdotoxin, but not Ba(2+) and ouabain. Prior stimulation with levcromakalim inhibited carbachol-induced responses, and this inhibitory effect was also sensitive to glibenclamide. 1, 3-dihydro-1-[2-hydroxy-5-(trifluoromethyl)phenyl]-5 (trifluoromethyl) -2H-benzimidazol-2-one (NS 1619), an activator of large conductance, Ca(2+)-activated K(+) channels (BK(Ca)), induced only small hyperpolarisations of the endothelial cells. Preincubation of tissues with 18 alpha- or 18 beta-glycyrrhetinic acid, inhibitors of gap junction communication, increased the input resistance and depolarised the endothelial cells, and inhibited the hyperpolarising effect of levcromakalim. It is concluded that activation of K(ATP) causes hyperpolarisation of rat mesenteric endothelial cells, probably through gap junctional transfer of smooth muscle hyperpolarisation, and that this may represent an important modulator of endothelial function. PMID- 10844126 TI - Reconstitution of functional dopamine D(2s) receptor by co-expression of amino- and carboxyl-terminal receptor fragments. AB - An N-terminal dopamine D(2s) receptor clone was constructed and coexpressed in COS-7 cells together with a separate gene fragment coding for the C-terminal sequence of the dopamine D(2s) receptor. The truncated receptor (referred to as D(2trunc)) contained transmembrane domains I-V and the N-terminal portion of the third cytoplasmic loop, whereas the C-terminal receptor fragment (referred to as D(2tail)) contained transmembrane domains VI and VII and the adjacent intra- and extracellular sequences of the dopamine D(2s) receptor. Expression in COS-7 cells of either of these two polypeptides alone did not result in any detectable [3H]methylspiperone binding activity. However, specific [3H]methylspiperone binding could be observed after coexpression of the D(2trunc) and D(2tail) gene constructs; the number of receptors present on the plasma membrane was about 10% with respect to that of the wild type. The binding properties of the coexpressed fragments were similar to those of the wild-type dopamine D(2s) receptor for agonists and antagonists. Functional stimulation of the cotransfected D(2trunc) and D(2tail) fragments with quinpirole resulted in the inhibition of adenylate cyclase activity. Maximal inhibition corresponds to a 28% decrease in forskolin stimulated adenylate cyclase. The apparent IC(50) of quinpirole was 5.1+/-0.3 mcM. These findings confirm and extend analogous data for other G protein-coupled receptors and indicate that this phenomenon is of general importance for the entire family of these proteins. PMID- 10844127 TI - Zolmitriptan stimulates a Ca(2+)-dependent K(+) current in C6 glioma cells stably expressing recombinant human 5-HT(1B) receptors. AB - Stimulation of a Ca(2+)-dependent K(+) current by zolmitriptan, a 5-HT(1B/1D) receptor partial agonist, was investigated in C6 glioma cells stably expressing recombinant human 5-HT(1B) receptors. Outward K(+) currents (I(K)) were examined in non-transfected C6 glioma cells and in cells expressing cloned human 5-HT(1B) receptors using the patch-clamp technique in the whole-cell configuration. In C6 glioma cells expressing recombinant human 5-HT(1B) receptor, zolmitriptan increased I(K) in a concentration-dependent manner (maximum increase 16.3+/-7.8%, n=5, p<0.001) with a pD(2) value (geometric mean with 95% confidence intervals) of 7.03 (7.90-6.10). Zolmitriptan failed to elicit increases in I(K) in non transfected C6 cells. In the presence of the mixed 5-HT(1B/1D) receptor antagonist, N-[4-methoxy-3-(4-methyl-1-piperazinyl)phenyl]-2(-methyl-4(5-methyl-1 ,2,4)-oxadiazol-3-yl)[1,1-biphenyl]-4-carboxamide 2HCl (GR 127935, 0. 1 mcM), zolmitriptan (1 mcM) failed to significantly increase I(K) in C6 cells expressing human 5-HT(1B) receptors confirming that zolmitriptan-evoked responses were indeed mediated by human 5-HT(1B) receptors. In C6 cells expressing cloned human 5-HT(1B) receptors, zolmitriptan-induced increases in I(K) were prevented by the calcium chelator, EGTA (5 mM) when included in the patch pipette (maximum increase -3.3+/-4.2%, n=4, P=NS). The Ca(2+)-dependent K(+) channel blockers, iberiotoxin (0.1 mcM) and tetraethylammonium (TEA, 1 mM), abolished zolmitriptan induced increases in I(K) (4.5+/-7.3%, n=4 and -0.8+/-1.7%, n=4, respectively, P=NS in each case) in C6 cells expressing human 5-HT(1B) receptors, confirming the involvement of Ca(2+)-dependent K(+) channels. In conclusion, the 5-HT(1B/1D) receptor partial agonist, zolmitriptan, stimulates I(K/Ca) in C6 glioma cells stably transfected with human 5-HT(1B) receptors suggesting an increase of hyperpolarizing current. PMID- 10844128 TI - Characterisation of the effects of SR146131, a novel non-peptide CCK(1) receptor agonist, on IMR-32 human neuroblastoma cells. AB - The effect of ?2-[4-(4-chloro-2, 5-dimethoxy-phenyl)-5-[2-cyclohexyl-ethyl) thiazol-2-ylcarbamoy l]-5, 7-dimethyl-indol-1-yl?-acetic acid (SR146131), a novel non-peptide agonist of cholecystokinin (CCK) CCK(1) receptors, was compared to the effect of sulphated cholecystokinin octapeptide (CCK-8-S) on CCK(1) receptors of the human neuroblastoma cell line IMR-32. SR146131 inhibited [125I]CCK-8-S binding to IMR-32 cells at nanomolar concentrations. SR146131 and CCK-8-S increased intracellular free Ca(2+) levels ([Ca(2+)](i)) in the same concentration range (EC(50)=6+/-2.3 and 1.3+/-0.14 nM, respectively). Although the shape of the [Ca(2+)](i) increase induced by CCK-8-S and SR146131 was slightly different, extracellular Ca(2+) removal affected the response of both compounds to a similar degree, and the response of both compounds was essentially due to Ca(2+) release from intracellular stores. This was also confirmed by measuring the [Ca(2+)](i) response of single cells: both compounds induced [Ca(2+)](i) oscillations at subnanomolar concentrations and elicited a large peak increase in [Ca(2+)](i) at higher concentrations (EC(50)=0.5+/-0.04 and 5.7+/-1.9 nM for CCK-8-S and SR146131, respectively). Both CCK-8-S and SR146131 induced a sustained increase of phosphoinositide turnover in these cells, and acted at similar concentrations (EC(50)=2.7+/-0.7 and 6+/-3.1 nM, respectively), although the maximal effect of SR146131 was somewhat lower than the effect of CCK-8-S. These data show that SR146131 activates human CCK(1) receptors on IMR-32 cells in a manner and with a potency similar to that of CCK-8-S. PMID- 10844129 TI - Changes in catecholaminergic pathways innervating the rat heart ventricle during morphine dependence. Involvement Of alpha(1)- and alpha(2)-adrenoceptors. AB - In the present study, we examined the effects of alpha(1)- and the alpha(2) adrenoceptors blockade on the changes in the ventricular content of catecholamines in rats withdrawn from morphine. Rats were given morphine by s.c. implantation of morphine pellets for 5 days. On the seventh day, morphine withdrawal was induced by s.c. administration of naloxone (1 mg/kg), and rats were killed 30 min later. Pretreatment with yohimbine (alpha(2)-adrenoceptor) or prazosin (alpha(1)-adrenoceptor) 15 min prior to naloxone administration attenuated some of the behavioural signs of morphine withdrawal. In addition, biochemical analysis indicated that yohimbine completely abolished the withdrawal induced increase in noradrenaline and dopamine turnover in the right ventricle. By contrast, prazosin did not block the hyperactivity of catecholaminergic neurons in the heart during withdrawal. These data suggest that the hyperactivity of catecholaminergic neurons in the heart during morphine withdrawal is dependent upon alpha(2)-adrenoceptor activation. In addition, the present results rule out the involvement of alpha(1)-adrenoceptors. PMID- 10844130 TI - Antinociceptive effects of tetrahydrocannabinol side chain analogs: dependence upon route of administration. AB - The role of flexibility of the alkyl side chain in the tetrahydrocannabinols to cannabinoid activity has been delineated in previous studies with side chain analogs of Delta(8)-tetrahydrocannabinol with double or triple bonds. This study investigated the site of antinociceptive action for these analogs through analysis of structure-activity relationships following different routes of administration. In analogs without terminal substitutions, potency was greater following intrathecal (i. t.) injection than with intracerebroventricular (i.c.v.). Further, optimal structural features differed for each route of administration. Absolute position of the double or triple bond best predicted i.t. potency. In contrast, i.c.v. potency was best predicted by the size of the alkyl substituent beyond the point of unsaturation. Terminal substitutions tended to increase i.c.v. potency while decreasing or not affecting i.t. These results suggest that receptor mechanisms for cannabinoid antinociceptive effects differ in brain and spinal cord, although potential pharmacokinetic differences in rate of local distribution cannot be eliminated. PMID- 10844131 TI - Neutral endopeptidase and alcohol consumption, experiments in neutral endopeptidase-deficient mice. AB - Alcohol consumption was investigated in mice which were rendered deficient in the peptide-degrading enzyme neutral endopeptidase (EC 3.4.24.11) (NEP-/-) by gene targeting and compared to alcohol consumption in corresponding wild type mice (NEP+/+). Mice were offered a free choice to drink tap water or 10% alcohol. The NEP-/- mice consumed significantly more alcohol ( approximately 42%) than the NEP+/+ mice, whereas no significant differences were observed in the total fluid consumption. The daily food consumption of alcohol naive NEP-/- animals was elevated ( approximately 29%). Furthermore, the activities of peptidases closely related to neutral endopeptidase were analysed ex vivo in several brain regions from NEP-/- and NEP+/+ mice not treated with alcohol. There was no obvious compensation for the total loss of neutral endopeptidase by the functionally related peptidases angiotensin-converting enzyme and aminopeptidase N. In vitro, the degradation of exogenously applied [Leu(5)]enkephalin was not reduced in membrane preparations of those brain regions assayed in NEP-/- mice. A small reduction in [Leu(5)]enkephalin degradation was detected in striatal membrane preparations of NEP-/- mice, if aminopeptidase N was additionally blocked by bestatin or amastatin. PMID- 10844132 TI - ATP-sensitive K(+) channel effects on nerve function, Na(+), K(+) ATPase, and glutathione in diabetic rats. AB - Some vasodilators correct nerve conduction velocity and endoneurial blood flow deficits in diabetic rats. It is not known whether vasa nervorum has ATP sensitive K(+) (K(ATP)) channels that mediate vasodilation, or whether K(ATP) channels could modulate peripheral nerve function. Therefore, we examined the effects of 2 weeks treatment with the K(ATP) channel openers, celikalim and WAY135201 (R-4-[3, 4-dioxo-2-(1, 2, 2-trimethyl-propylamino)-cyclobut-1-1 enylamino]-3-methoxy-+ ++benzonitri le), on sciatic nerve blood flow, conduction velocity, Na(+)-K(+) ATPase activity and glutathione content after 6 weeks of untreated streptozotocin-diabetes in rats. Blood flow and motor conduction velocity, 47.6% and 20.3% reduced by diabetes, respectively, were completely restored by both celikalim and WAY135201 treatments. Diabetes diminished sciatic Na(+)-K(+) ATPase activity by 47.6% and this was 80-90% corrected by the K(ATP) channel openers. Sciatic nerve glutathione content, 30.3% reduced by diabetes, was unaffected by celikalim or WAY135201. Thus, K(ATP) channel openers had marked beneficial effects on nerve perfusion and function in experimental diabetic neuropathy, and may be suitable for further study in clinical trials. PMID- 10844133 TI - Paradoxical aggravation of paroxysmal dystonia during chronic treatment with phenobarbital in a genetic rodent model. AB - Recent studies in mutant hamsters (dt(sz)), an animal model of primary paroxysmal dystonia, indicated that altered function of the gamma-aminobutyric acid (GABA)ergic system plays a critical role in the pathogenesis of dystonia. In the present study, dt(sz) hamsters were chronically treated with phenobarbital, which has been found to exert antidystonic effects in mutant hamsters after acute administration. In untreated dt(sz) hamsters, the severity of dystonia follows an age-dependent time course with a maximum between the 30th and 40th day of life, followed by a continuous decline of severity until complete remission occurs at the age of about 70 days. In contrast to acute effects, chronic treatment with phenobarbital via drinking water starting at an age of 21 days (i.e., after weaning) worsened dystonia and retarded the spontaneous remission. The unexpected prodystonic effect was more marked after administration of higher doses and when chronic treatment with phenobarbital started at an age of 1 day (neonatal administration via breast milk). After withdrawal of phenobarbital at the age of 70 days, the severity rapidly declined in all treated groups. When phenobarbital was readministered 1 week later, the hamsters again exhibited severe dystonia. The mechanism of these unexpected findings is unknown. Tentatively, activity dependent GABA-mediated excitation caused by chronic treatment with phenobarbital may be important for the prodystonic effects under pathological conditions in dt(sz) hamsters. PMID- 10844134 TI - Desensitisation of 5-HT autoreceptors upon pharmacokinetically monitored chronic treatment with citalopram. AB - Rats were chronically treated with the selective serotonin re-uptake inhibitor citalopram [1-(3-dimethylaminopropyl)-1-(4-fluorophenyl)-5-phtalancarbonitril ], by means of osmotic minipumps. Using an infusion concentration of 50 mg/ml citalopram, steady-state plasma concentrations of approximately 0.3 mcM citalopram were maintained for 15 days. Citalopram plasma levels dropped below pharmacologically active concentrations 48 h after removal of the minipumps. Although chronic treatment with citalopram did induce an attenuated response by extracellular levels of 5-hydroxytryptamine (5-HT) after systemic administration of the 5-HT(1A) receptor agonist 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH DPAT), no effect of chronic citalopram treatment was observed when 5-HT(1B) receptor function was evaluated with a local infusion of 5-HT(1B/D) receptor agonist, sumatriptan (3-[2-dimethylamino]ethyl-N-methyl-1H-indole-5methane sulphonamide). Controversially, no augmentation of the increase of 5-HT levels was observed upon systemic administration of citalopram. It is concluded that, although chronic treatment with citalopram does induce desensitisation of 5 HT(1A) receptors, the absence of augmented effects of citalopram on 5-HT levels indicates that other mechanisms compensate for the loss of autoreceptor control. PMID- 10844135 TI - Effect of trandolapril on vascular responsiveness in cholesterol-fed rabbit isolated arteries. AB - According to the World Health Organisation, cardiovascular disorders are one of the main causes of morbi/mortality in the western world. The effect of trandolapril (0.3 mg kg(-1) day(-1)), a non-sulphydryl angiotensin-converting enzyme (ACE) inhibitor, on the vascular responsiveness in aorta isolated from hypercholesterolemic rabbits was examined. Three groups of rabbits (n=30) were used: Group 0 (control group); Group 1 (hypercholesterolemic group, 0.5% (wt/wt) cholesterol-enriched diet) and Group 2 (hypercholesterolemic+trandolapril 0.3 mg kg(-1) day(-1)). After 18 weeks of treatment, the rabbits were killed and the thoracic aorta, proximal coronary and mesenteric (5th branch) arteries were isolated, cleaned off and mounted in an organ bath. Trandolapril had no significant effect on plasma cholesterol, high density lipoprotein (HDL) or low density lipoprotein (LDL). Despite the lack of effect of the drug on the above mentioned parameters, treatment with trandolapril improved endothelium-dependent relaxation induced by acetylcholine in aortic and mesenteric rings from hypercholesterolemic rabbits treated with trandolapril. The relaxation induced by 10(-5) M acetylcholine were 65.0+/-4.0%; 24. 0+/-9.4% (P<0.01, n=10) and 51.3+/ 7.0% (P<0.01, n=10) in aortic rings from Groups 0, 1 and 2, respectively, and 50.0+/-12.0%; 10. 1+/-10.0% (P<0.01, n=10); 61.0+/-9.7% (P<0.01, n=10) in small mesenteric rings from Groups 0, 1 and 2, respectively. In addition, trandolapril treatment improved the increase in serotonin-induced contraction in proximal coronary arteries with respect to the hypercholesterolemic group. On the other hand, we did not find any differences among the group in endothelium-independent relaxation induced by sodium nitroprusside. These results provide evidence that trandolapril restores endothelium-dependent relaxation in hypercholesterolemic rabbit-isolated arteries. These data suggest that trandolapril might have beneficial action in the prevention of vascular alteration involved in atherosclerosis. PMID- 10844136 TI - Endothelin-1 attenuates bradykinin-induced hypotension in rats. AB - Endothelin-1 has vasoconstrictor and mitogenic properties and may contribute to the pathogenesis of hypertension by enhancing vasoconstrictor mechanisms. In this study, we investigated the ability of endothelin-1 decrease the hypotensive effects of the vasodilator bradykinin in anesthetized rats. We also studied the effects a two-week oral pre-treatment with losartan (10 mg/kg/day) or enalapril (25 mg/kg/day) on endothelin-1-induced changes in the hypotensive responses to bradykinin. Bradykinin (0.4, 1.6, 6.4, and 25 mcg/kg, i.v.) induced dose dependent hypotensive responses which were attenuated (P<0.05) by endothelin-1 (2 mcg/kg, i.v.). This effect of endothelin-1 was abolished by the mixed endothelin receptor antagonist N-Acetyl-alpha-[10,11-Dihydro-5H-dibenzo[a, d]cycloheptadien 5-yl]-D-Gly-Leu-Asp-Ile-Ile-Trp (PD145065, 1 mg/kg, i.v.). Endothelin-1 also decreased (P<0.05) the responses to bradykinin in rats pre-treated with losartan, but had no effect in rats pre-treated with enalapril. These results suggest that endothelin-1 may contribute to the development of hypertension by decreasing the responses to bradykinin through a mechanism not involving angiotensin AT(1) receptors, although the inhibition of angiotensin converting enzyme blunted the effect of endothelin-1. PMID- 10844137 TI - Positive inotropy mediated via CGRP receptors in isolated human myocardial trabeculae. AB - Isometric contractile force were studied on isolated human myocardial trabeculae that were paced at 1.0 Hz in tissue baths. Alpha calcitonin gene-related peptide (alpha-CGRP) had a potent positive inotropic effect in most trabeculae from both the right atrium and left ventricle, and this effect was partially antagonized by the CGRP(1) receptor antagonist alpha-CGRP-(8-37) (10(-6) M). Amylin and the CGRP(2) receptor agonist [Cys(acetylmethoxy)(2, 7)]CGRP had a positive inotropic effect in some trabeculae, whereas adrenomedullin had no inotropic effect. Using reverse transcriptase-polymerase chain reaction (PCR) mRNAs encoding the human calcitonin receptor-like receptor and the receptor associated modifying proteins (RAMPs) RAMP1, RAMP2, and RAMP3 were detected in human myocardial trabeculae from both the right atrium and left ventricle. In conclusion, functional CGRP(1) and CGRP(2) receptors may mediate a positive inotropic effect at both the atrial and ventricular level of the human heart. mRNAs for calcitonin receptor-like receptor and specific RAMPs further support the presence of CGRP receptors. PMID- 10844138 TI - [Nphe(1)]nociceptin-(1-13)NH(2) selectively antagonizes nociceptin effects in the rabbit isolated ileum. AB - When suspended in vitro in isolated organ baths, segments of the rabbit ileum show a fairly strong and stable spontaneous activity, which derives from the continuous release of acetylcholine and the activation of muscarinic receptors, since the activity is completely eliminated by atropine. Dynorphin A (pEC(50): 8.6+/-0.07), neuropeptide Y and its congener human pancreatic polypeptide (pEC(50): 9.40+/-0.10), and nociceptin (pEC(50): 8.08+/-0.12) dose-dependently inhibit the spontaneous activity through the activation of receptors, which are specifically antagonised respectively by naloxone (pA(2): 7.17+/-0.12), 2 (naphtalen-1-ylamino)-3-phenylpropionitrile (JCF 104; pA(2): 5. 80+/-0.10), and [Nphe(1)]nociceptin-(1-13)NH(2) (pA(2): 6.17+/-0.19). This last compound, a selective nociceptin-receptor (OP(4)) antagonist, inhibits the effect of nociceptin in a competitive manner, as demonstrated by Schild analysis. [Nphe(1)]nociceptin-(1-13)NH(2) also antagonizes the effects of other OP(4) receptor ligands such as the full agonist, nociceptin-(1-13)-NH(2), and the partial agonists, [Phe(1)psi(CH(2)-NH)Gly(2)]nociceptin-(1-13)-NH(2) (intrinsic activity (alpha(E))=0.5) and Ac-RYYWK-NH(2) (alpha(E)=0.5), with pA(2) values ranged from 5.8 to 6.2. These results indicate that the functional site mediating the inhibitory effect of nociceptin in the rabbit ileum, is pharmacologically identical to the OP(4) sites of other species (mouse, rat, guinea pig, man), since the potencies (pA(2) values) of the pure and competitive antagonist [Nphe(1)]nociceptin-(1-13)NH(2) is very similar to the values obtained in the other species. Moreover, the rabbit ileum is one of the few isolated organs that allow classifying compounds, which interact with OP(4) receptors as full agonists, partial agonists, or pure antagonists. PMID- 10844140 TI - Corrigendum to: "Noradrenergic lesion antagonizes desipramine-induced adaptation of NMDA receptors" PMID- 10844139 TI - Interplay between nitric oxide and vasoactive intestinal polypeptide in the pig gastric fundus smooth muscle. AB - The aim of this study was to investigate the exact mechanism of interaction between nitric oxide (NO) and vasoactive intestinal polypeptide (VIP) as inhibitory non-adrenergic non-cholinergic (NANC) neurotransmitters in isolated smooth muscle cells and smooth muscle strips of the pig gastric fundus. In isolated smooth muscle cells, the maximal relaxant effect of VIP (10(-9) M) was inhibited by 94% by the NO synthase (NOS) inhibitor N(G)-nitro-L-arginine (L-NA, 10(-4) M) and by 85% by the inducible NOS (iNOS)-selective inhibitor N-(3 (aminomethyl)-benzyl)acetamide (1400W; 10(-6) M). The relaxant effect of VIP was reduced by more than 70% by the guanylyl cyclase inhibitor 1H (1,2,4)oxadiazolo(4,3-a)quinoxalin-1-one (ODQ; 10(-6) M), the glucocorticoid dexamethasone (10(-5) M) and three protein kinase A inhibitors: (R)-p-cyclic adenosine-3', 5'-monophosphothioate ((R)-p-cAMPS; 10(-6) M), ?(8R,9S, 11S)-(-)-9 hydroxy-9-n-hexylester-8-methyl-2,3,9,10-tetrahydro-8, 11-epoxy-1H,8H,11H 2,7b,11a-triazadibenzo[a, g]cycloocta[cde]-trin-den-1-one? (KT5720; 10(-6) M) and N-(2-(p-bromo-cinnamylamino)ethyl))-5-isoquinoline sulfonamide dihydrochloride (H 89; 10(-5) M). In contrast, no influence of the NOS inhibitors, ODQ, dexamethasone, nor the protein kinase A inhibitors could be observed on the relaxant effect of VIP in smooth muscle strips. These data demonstrate that the experimental method completely changes the influence of NOS inhibitors on the relaxant effect of VIP in the pig gastric fundus. The isolation procedure of the smooth muscle cells might induce iNOS that can be activated by VIP. PMID- 10844142 TI - Vitellogenic ovarian follicles of Drosophila exhibit a charge-dependent distribution of endogenous soluble proteins. AB - In ovarian follicles of Drosophila, soluble endogenous charged proteins are asymmetrically distributed dependent upon their ionic charge. Reversal of the normal ionic difference across the intercellular bridges which connect nurse cells to their oocyte results in a redistribution of these proteins. Twelve soluble endogenous acidic proteins were identified by 2-D gel electrophoresis as being present in both oocytes and nurse cells in samples run on four or more gels. Of these, following osmotically induced reversal of the electrical transbridge gradient the concentration of seven proteins decreased in the oocyte while nurse cell concentrations of all twelve proteins increased. Of seven basic proteins analyzed, following reversal of the electrical gradient the concentration of all seven increased in oocytes. Four of these decreased in nurse cells, while nurse cell concentrations of the remaining three basic proteins also appeared to decrease, but yielded spots too faint for measurement. Data presented here demonstrate that, as in the Saturniidae, the ionic gradient across the nurse cell-oocyte intercellular bridges of the dipteran, Drosophila, can influence the distribution of soluble endogenous charged molecules. PMID- 10844141 TI - Altered behaviour following RNA interference knockdown of a C. elegans G-protein coupled receptor by ingested double stranded RNA. AB - Using a systemic and continuous delivery method based on feeding on a particular strain of transformed Escherichia coli to induce double stranded RNA-mediated interference, we targeted the product of the npr-1 gene, a putative Caenorhabditis elegans homologue of a neuropeptide Y receptor, a G-protein coupled receptor. We were able to reproduce the social behaviour observed for the naturally occurring npr-1 mutant when wild type N2 Bristol eggs developed in a lawn of bacteria producing double stranded RNA for npr-1. This facile approach may also be useful when studying the function of other worm G-protein coupled receptors. PMID- 10844143 TI - Acid phosphatases in the haemolymph of the desert locust, Schistocerca gregaria, infected with the entomopathogenic fungus Metarhizium anisopliae. AB - A comparison has been made between the effects of wounding, chemical stimulation of the immune system and fungal infection on acid phosphatase (AcP) activity in the haemolymph of the desert locust, Schistocerca gregaria. Untreated control locusts had constitutive levels of AcP. As a lysosomal enzyme, AcP may have a role in autophagy and cell turn over as well as defence. Injection of saline and beta-1,3-glucan caused significant increases in haemocyte and plasma AcP. AcP activity also increased in the haemolymph on the 3rd day after inoculation with the entomopathogenic fungus M. anisopliae var acridum. This coincided with a decline in the total haemocyte count and a marked reduction in the proportion of plasmatocytes and coagulocytes that stained positive for AcP. Therefore a priori it seemed unlikely that the extra AcP in infected insects came from the host. A fungal origin for the enzyme was suggested by the identification of AcP isoforms from haemolymph of different treatments. Control inoculated (oil only) insects had an AcP at a pI of 4.3 that was stimulated further by the injection of laminarin. Additional isoforms appeared at around 7.3-7.5 in the laminarin treatment. However, the 4.3 isoform appeared to be suppressed in the insects infected with M. anisopliae var acridum. The band intensity was more like that of the control than the laminarin-injected insects. Two new isoforms appeared later on in infection. These enzymes had pIs that corresponded to some of the AcPs produced in vitro by the fungus. The results are discussed in the light of the possible benefits of secreted fungal acid phosphatases to the pathogen. PMID- 10844144 TI - Sulfakinins reduce food intake in the desert locust, Schistocerca gregaria. AB - In vertebrates, the peptides cholecystokinin (CCK), neuropeptide Y, galanin, and bombesin are known to be involved in the control of food intake. We report here that insect sulfakinins, peptides which display substantial sequence similarities with the vertebrate gastrin/CCK peptide family, significantly inhibit food uptake in fifth instar nymphs of the locust, Schistocerca gregaria. Upon injection of Lom-sulfakinin, a neuropeptide present in the corpus cardiacum of locusts, food intake was significantly reduced in a dose-dependent manner within a fixed 20 min time period. The induced effect ranged from 13% inhibition (10 pmol of injected peptide) to over 50% inhibition at 1 nmol. Other naturally occurring sulfakinins from different insect species also elicited this satiety effect. Analogous to the satiety effect of CCK in vertebrates, the sulfate group is required for activity. No effect on the palptip resistance was found after injection with sulfakinin. Therefore it seems unlikly that sulfakinins reduce food intake by decreasing the sensitivity of the taste receptors. PMID- 10844145 TI - Possible bases of pseudoparasitism in Spodoptera littoralis larvae stung by Microplitis rufiventris. AB - The effects of host age and parasitoid female age on the occurrence of 'Pseudoparasitism', using the Spodoptera littoralis-Microplitis rufiventris host parasitoid system were investigated. The first four larval instars of the host are not equally suitable for parasitoid development. The proportion of pseudoparasitized hosts significantly increases when: (1) the age of the female parasitoid increases; (2) oviposition occurs mostly in fourth instar larvae; (3) a later age of the host instar is used; (4) the mandibles of the newly hatched parasitoid larvae mistakenly attack host interior organs (e.g. Malpighian tubules); and (5) an imperfect growth pattern of teratocytes occurs. The reluctance of female wasps to parasitize fourth instar host larvae is not due to the thickness of host cuticle but possibly due to the unfavourable physiological state of the host larvae. The age of host larvae at the time of parasitization may influence the adverse effects of parasitoid factors (e.g. polydnavirus, venom and teratocytes) on the growth of host larvae. It is suggested that females of M. rufiventris are able to determine the suitability of a potential host instar for the development of their offspring. The cell diameter of M. rufiventris teratocytes increases with increasing age of host larvae at the time of oviposition. The association within the host of living parasitoid larvae and functional teratocytes may be important for the survival of each other and consequently for successful parasitism. PMID- 10844146 TI - Evidence for serine protease inhibitor activity in the ovarian calyx fluid of the endoparasitoid Venturia canescens. AB - Endoparasitic wasps are able to develop inside permissive host insects due to their ability to overcome or evade the host's immune system. In the present study, we provide experimental evidence that ovarian calyx fluid of the ichneumonid endoparasitoid Venturia canescens has the potential to alter host haemocyte spreading and inhibit host haemolymph melanisation due to the presence of a putative serine protease inhibitor (serpin) activity. The existance of a serpin-like activity in the calyx fluid is also supported by experiments where the synthetic protease inhibitor p-APMSF had effects on cellular and cell-free immune reactions similar to ovarian calyx fluid. In addition, based on proteolytic digestion patterns of a wasp egg surface protein, we predict an Arg specific trypsin-like protease activity in the host haemolymph which is possibly affected by calyx fluid components as well. Our data suggest that ovarian calyx fluid, deposited into the host together with the parasitoid egg, contains serpin activity which might transiently inactivate host defence reactions until other means of protection are established on the egg surface. PMID- 10844147 TI - Typical ventilatory pattern of the intact locust is produced by the isolated CNS. AB - Ventilatory rhythms of locusts are generated in the central nervous system (CNS). The primary oscillator or central pattern generator (CPG) is located in the metathoracic ganglion. We studied the different patterns of ventilation by recording long-term efferent discharges from the isolated metathoracic ganglion.Two different basic patterns occur: continuous ventilation and discontinuous ventilation. These patterns can be found in the isolated nerve cord as well as in intact animals. In intact animals sensory feedback usually elicits high frequency continuous ventilation as is the case in most physiological experiments. Many studies of ventilation-associated interneurones were performed under what we call stressed conditions i.e. with strong sensory feedback. Under these conditions many interneurones may be recruited which probably do not belong to the basic CPG. In isolated nerve cords of locusts we recognised the two basic types of ventilation. This provides an experimental approach to the origin of rhythmogenesis in ventilation. We can now examine single interneurones under less stressed or even discontinuous ventilatory conditions in the isolated CNS.We suggest the dominance of intrinsic rhythmogenesis of ventilation in the metathoracic ganglion of locusts. PMID- 10844148 TI - The influence of mechanical, visual and contact chemical stimulation on the behavioural phase state of solitarious desert locusts (Schistocerca gregaria). AB - We investigated the influence of mechanical, visual and contact chemical stimulation on behavioural gregarization of fifth-instar solitarious nymphs of the desert locust. The stimuli were applied in two 2x2 factorial experimental designs, the first with contact chemical and mechanical stimuli, and the second with contact chemical and visual stimulation. Stimulus treatments were applied for a 4-5 h period, after which the behavioural phase state of individual locusts was measured using an assay based on multiple logistic regression analysis of behavioural variables. Mechanical stimulation was provided by showering test insects with millet seeds, thereby excluding the possibility of contact chemical self-stimulation by repeated contact with the same objects. Visual stimulation consisted of the sight of crowd-reared locusts, while contact chemical stimulation was a dichloromethane extract of cuticular hydrocarbons from gregarious nymphs applied to the perch of the test insect. Mechanical stimulation was powerfully gregarizing, whether alone or in combination with contact chemical stimuli. Application of cuticular extract to the perch had no measurable effect on behavioural phase state, either alone or when presented with mechanical or visual stimuli. Visual stimulation alone partly gregarized test locusts. These results appear to conflict with other reports of the gregarizing effect of cuticular hydrocarbons and possible reasons for this discrepancy are discussed. PMID- 10844149 TI - The importance of the spermathecal duct in bumblebees. AB - The elongated spermathecal duct of bumblebees has been studied in hibernating queens, queens shortly after hibernation, mature egg-laying queens and uninseminated queens captured during summer, and workers. Only rather small size differences are found when comparing spermathecae of queens and workers. Clear differences between bumblebee queens and workers are found when comparing the histochemistry of the spermathecal ducts. Adult queens, regardless of age and reproductive status have spermathecal ducts that contain PAS positive material, whereas workers do not. It is suggested that the polysaccharides in the spermathecal ducts of queens are necessary as a source of energy for the rapid activation of spermatozoa passing through the duct prior to oocyte fertilization. An ultrastructural investigation revealed the presence of high glycogen content in the cells lining the duct of queens. Assuming that sperm cells are kept in a rather inactive state in the reservoir, the carbohydrate (glycogen) probably serves as an energy source for the sperm. The comparatively increased spermathecal duct length of bumblebees may increase the retention time of sperm inside the lumen. PMID- 10844150 TI - Permeability and disruption of the peritrophic matrix and caecal membrane from Aedes aegypti and Anopheles gambiae mosquito larvae. AB - In mosquito larvae, the peritrophic matrix (PM) separates the gut contents from the intestinal epithelium. This report describes a new in vivo assay for estimating PM permeability. The assay also allows for assessment of the permeability of the caecal membrane, a structure that separates each caecum from the gut lumen. Permeability was estimated by the appearance of fluorescently labeled dextrans (size range 4,400 to 2 million Da) within the gastric caecae of mosquito larvae. While the intact peritrophic matrix was impermeable to 2 million Da dextran particles, it was permeable to dextran particles of 148 kDa and smaller. The caecal membrane appears to have considerably smaller pores, being permeable only to dextrans of 19.5 kDa and smaller. The assay was also used to devise a treatment that disrupts the PM sufficiently to allow the passage of virus-sized particles. Dithiothreitol and to a lesser extent, chitinase were effective in disrupting the PM. Cycloheximide had a small effect; Polyoxin D, Pronase and calcofluor did not alter the permeability to 2 million Da dextran particles. Disruption of the PM is discussed in the context of infecting mosquitoes with retroviral transformation vectors. PMID- 10844151 TI - Overwintering strategy in Pyrrhocoris apterus (Heteroptera): the relations between life-cycle, chill tolerance and physiological adjustments. AB - Seasonal dynamics of ecophysiological parameters are described which are relevant to overwintering in field-collected adults of a Czech population of the red firebug, Pyrrhocoris apterus. Five life-cycle phases were distinguished using the duration of pre-oviposition period as a criterion: reproductive activity (spring early summer), intensification of reproductive diapause (RD) (peak of summer), maintenance of RD (late summer-early autumn), termination of RD (late autumn early winter), and low temperature quiescence (LTQ) (winter). The supercooling capacity and chill tolerance (c.t.) increased simultaneously with the termination of RD and all three processes were triggered/conditioned by autumnal decrease in ambient temperatures. Maximum supercooling capacity and c.t. 'outlived' the end of diapause and persisted throughout the LTQ state. The limits of c.t. were estimated as -15 degrees C/1-2 weeks for 50% survival. Ribitol, sorbitol, arabinitol, and mannitol were accumulated in the winter-sampled insects. Relatively low concentrations of polyols (dominating ribitol reached ca. 1% FW) indicate that they do not function as colligative cryoprotectants. However, because their seasonal occurrence coincided with the highest c.t., their non colligative cryoprotectant effects would merit further study. Although the overwintering microhabitat of P. apterus is buffered, the temperatures may fall to -13 degrees C during exceptionally cold winters and thus, the parameters of c.t. seem to be just appropriately tuned to the local overwintering conditions. PMID- 10844152 TI - Behaviour of 3-week weaned pigs in Straw-Flow(R), deep straw and flatdeck housing systems. AB - The behaviour of 3-week weaned pigs in different housing systems was examined as part of an assessment of the suitability of the Straw-Flow(R) system for pigs of this age. Three replicate pens of 20 pigs were weaned at 6.4 kg liveweight into each of: (a) deep-straw; (b) Straw-Flow(R); (c) large flatdeck; (d) small flatdeck. A kenneled lying area was provided in (a) and (b). The floor in (c) and (d) was expanded metal. Stocking densities were 0.23 m(2)/pig in (a), (b) and (c), and 0.17 m(2)/pig in (d). After 4-5 weeks (at 19.6 kg liveweight), 16 pigs from each pen were moved into Straw-Flow(R) grower pens (0.68 m(2)/pig) and observed until slaughter at 90.6 kg. Pigs in systems incorporating straw showed behaviour patterns associated with increased welfare (greater straw-directed behaviour and less pig-directed and pen-directed behaviour) relative to those in barren pens. Behavioural differences between (a) and (b) related to differences in available straw; there were few differences between (c) and (d). Pigs from (c) and (d) showed increased rooting relative to those from (a) after transfer to the grower pens, but other behavioural differences between weaner treatments did not persist. It is concluded that the Straw-Flow(R) system can provide suitable accommodation for weaned pigs. PMID- 10844153 TI - Behavioural interactions between West African dwarf nanny goats and their twin born kids during the first 48 h post-partum. AB - West African dwarf nanny goats and their twin-born kids were tested to determine their behavioural response to separation and their mutual recognition during the first 48 h post-partum. Does and their kids were given scores ranging from 1 to 5, depending on how they performed in the tests. Animals that showed maximum response and recognition ability were given a score of 5, while those with minimum response and recognition ability scored 1. The kids were prevented from sucking 2 h prior to the tests, which were carried out at 18, 24, 36 and 48 h post-partum. Chi-square procedure was used to determine whether age, sex and birthweight of kids as well as hours post-partum and parity of dams had any effect on these post-partum behaviours. Out of 48 twin-born kids tested, 32 (67%) responded actively to separation from dams (i.e. had scores of 3 or more). The age, sex and birthweight of kids did not significantly affect (P>0.05) their response to separation from their dams. The hours post-partum and the parity of does also did not affect their response to separation from their kids. The dam recognition ability of twin-born kids was very poor. Out of a total of 48 kids tested, only 17 (35%) were able to recognize their dams (i.e. had scores of 3 or more). Even at 36 h, only four out of 14 (26%) could recognize their dams. It was only at 48 h that the majority of kids tested (i.e. 75%) successfully identified their dams. At 48 h, the dam recognition ability of kids was significantly better (P<0.05) than that of 18-h-old kids. Sex and birthweight of 24-48-h-old twin-born kids did not significantly affect (P>0.05) their ability to recognize their dams. The majority of does tested (i.e. 20 out of 24) were able to recognize their twin born kids. The hour post-partum and parity of does did not significantly affect (P>0.05) their kid recognition ability. PMID- 10844154 TI - Lambs fed protein or energy imbalanced diets forage in locations and on foods that rectify imbalances. AB - Ruminants eat a variety of foods from different locations in the environment. While water, cover, social interactions, and predators are all likely to influence choice of foraging location, differences in macronutrient content among forages may also cause ruminants to forage in different locations even during a meal. We hypothesized that lambs forage at locations containing foods that complement their basal diet and meet their nutritional needs. Based on this hypothesis, we predicted that lambs (n=12) fed a basal diet low in protein and high in energy would forage where a high-protein food (Food P) was located, and that lambs (n=12) fed a basal diet low in energy and high in protein would forage where a high-energy food (Food E) was located. Food P was a ground mixture of blood meal (50%), grape pomace (30%), and alfalfa (20%) that contained 47% crude protein (CP) and 2.211 Mcal/kg digestible energy (DE). Food E was a ground mixture of cornstarch (50%), grape pomace (30%), and rolled barley (20%) that contained 6% CP and 3.07 Mcal/kg DE. Food P provided 212 g CP/Mcal DE, whereas Food E provided 20 g CP/Mcal DE. Lambs growing at a moderate rate require 179 g CP and 3.95 Mcal DE. During Trial 1, we determined if lambs foraged to correct a nutrient imbalance, and if they preferred a variety of foods (Foods P and E) to only one food at a location (Food P or E). During Trial 2, we determined if nutrient-imbalanced lambs foraged in the location with the food that corrected the imbalance when the location of the foods changed daily. During Trial 3, lambs were offered familiar foods (Foods P and E) at the location furthest - and novel foods (wheat and soybean meal) at the location nearest - the shelter of their pen. During all three trials, lambs foraged most at the location with the food that contained the highest concentration of the macronutrient lacking in their basal diet, but they always ate some of both foods. Lambs did not feed exclusively at the location with a variety of foods (P and E). Rather, they fed at the location nearest the shelter that contained the macronutrient lacking in their diet. As availability of the food with the needed macronutrient declined in one location, lambs moved to the nearest location that had food with the needed macronutrient. When food that complemented their basal diet was moved to a different location, lambs foraged in the new location. Collectively, these results show that lambs challenged by imbalances in energy or protein selected foods and foraging locations that complemented the nutrient content of their macronutrient imbalanced basal diets. PMID- 10844155 TI - The preferences of laying hens for different concentrations of atmospheric ammonia. AB - Ammonia gas is one of the most abundant aerial pollutants of modern poultry buildings. The current chronic exposure limit for ammonia of 25 ppm is set for human safety rather than animal welfare. This study assessed the behavioural preferences of laying hens (Gallus gallus domesticus) for different concentrations of ammonia found in commercial poultry houses. Six groups, each of six laying hens, were given the choice of three concentrations of ammonia ( approximately 0, 25 and 45 ppm) in a preference chamber over a period of 6 days and their location and behaviour recorded every 15 min. Hens foraged (p=0.018), preened (p=0.009) and rested (p=0.029) significantly more in fresh air than in the ammonia-polluted environments. There was a significant difference between the responses in 0 and 25 ppm (p<0.05) but not between 25 and 45 ppm (p>0.05). This suggests that ammonia may be aversive to hens with a threshold for this aversion between 0 and 25 ppm. Future studies should explore graded concentrations of ammonia between 0 and 25 ppm in order to suggest a new chronic exposure limit on the basis of animal welfare. PMID- 10844156 TI - Aggression and social spacing in light horse (Equus caballus) mares and foals. AB - Aggression and social spacing were studied in 14 light horse mares and their foals living at pasture. Focal samples were collected on each mare-foal dyad for 6 to 10.5 h from 2 months of foal age until weaning at approximately 4 months of age. Observations on foals continued until approximately 6 months of age for 7.5 to 10.5 h per foal. Every 2 min the identities of all individuals within 5 m were recorded. All occurrences of agonistic behavior, and the participants, were recorded during the focal samples. In addition, during feeding of supplemental grain, all occurrences of agonistic behavior by all subjects were recorded. Significant correlations were found between mare rank and the rank of foals both prior to and after weaning. Before weaning, the rank of the foal was significantly correlated with birth order. No significant correlation between birth order and foal rank was found for the post-weaning hierarchy. An animal's gender had no significant effect on foal rank or the choice of preferred associate. Both prior to and after weaning, foals associated preferentially with the foal of their dam's most preferred associate. In addition, significant positive correlations were found between rank of mares and foals and the rate at which they directed aggression to other herd members. PMID- 10844157 TI - Indication of a genetic basis of stereotypies in laboratory-bred bank voles (Clethrionomys glareolus). AB - The development of stereotypies was studied in two successive laboratory-bred generations of bank voles representing F1 (n=248) and F2 (n=270) of an originally wild caught stock. It was shown that the propensity to develop stereotypies under barren housing conditions strongly relates to the same propensity of the parents. Stereotypies were approximately seven times more frequent in the offspring of stereotyping parents than in the offspring of permanent non-stereotypers. This held true even when only one of the parents was stereotyper. The paternal and maternal contributions to stereotypies in the offspring appeared to be equal. Males showing stereotypies but prevented from any physical contact with the offspring were as potent as stereotyping females in producing stereotyping offspring. Moreover, the specific type of stereotypy appearing in the offspring after isolation was very much related to the type of stereotypy developed in the mothers. We found no support for the possible importance of social facilitation from littermates, in that the development of stereotypies was independent of the length of time the voles were kept socially with littermates before isolation. We suggest that the possible genetic basis of individual differences in the propensity to develop stereotypies in captivity may result from differences in genetic predispositions and their interactions with discrete frustrating stimuli early in life and/or to genetically different predispositions to cope with frustrating experiences later in life. PMID- 10844158 TI - Development of stereotypies and polydipsia in wild caught bank voles (Clethrionomys glareolus) and their laboratory-bred offspring. Is polydipsia a symptom of diabetes mellitus? AB - The development of stereotypies and polydipsia was studied in wild caught bank voles (P: n=92) and their laboratory-bred offspring (F1: n=248). All animals were kept isolated in barren cages in the laboratory. In the P generation, no individuals developed stereotypies, but 22% developed polydipsia (>21 ml/day water intake against normally 10 ml/day). Polydipsia was more frequent among males (34%) than females (13%). In F1, 30% developed locomotor stereotypies alone, 21% showed polydipsia alone, and, additionally, 7% developed both stereotypies and polydipsia. Fewer males than females developed stereotypies (23% vs. 38%), whereas polydipsia was more frequent in males than in females (30% vs. 11%). The occurrence and distribution of polydipsia among sexes were the same in F1 and P. The distribution of different types of stereotypies in stereotyping voles were backward somersaulting (BS, 80%), high-speed jumping (JUMP, 29%), pacing following a fixed route (PF, 12%) and windscreen wiper movement (WIN, 5%). Some individuals (10%) showed two or more different types of stereotypies. The average age for developing stereotypies was 96 days while polydipsia was registered at the age of 63 days in both sexes. Voles showing both polydipsia and stereotypies developed polydipsia later (79 days) than polydipsic voles not showing stereotypies. This difference was especially pronounced in stereotyping females in which the occurrence of polydipsia was postponed to the age of 114 days. Polydipsic voles were tested positive for glucosuria indicating that polydipsia could be a symptom of diabetes mellitus. It is suggested that the development of stereotypies and polydipsia among bank voles in the laboratory are the results of frustration and prolonged stress. Stereotypies seem to depend on frustrative experiences early in life, while polydipsia may be related to diabetes mellitus caused by the experience of prolonged stress. Moreover, circumstances related to the development of stereotypies may be adaptive by reducing the risks of prolonged stress, including the development of fatal polydipsia. PMID- 10844160 TI - A convenient method for the determination of the quality of goldenseal. AB - Goldenseal (Hydrastis canadensis) has emerged as one of the top five herbal supplements in the world-wide market. A convenient method for the determination of the quality and possible adulteration of goldenseal products is thin-layer chromatography (TLC). TLC analysis of 10 goldenseal samples using two solvent systems and spray reagents was conducted. Five of these samples contained both hydrastine and berberine, four contained berberine and one did not contain either of these alkaloids. These TLC results were verified by HPLC analysis. PMID- 10844159 TI - Immunomodulatory and antitumour properties of Psoralea corylifolia seeds. AB - Psoralea corylifolia seed extract has been found to stimulate the immune system in mice. Administration of the extract was found to inhibit EAC ascitic tumour growth and stimulate natural killer cell activity, antibody-dependent cellular cytotoxicity, antibody-forming cells and the antibody complement-mediated cytotoxicity during tumour development. PMID- 10844162 TI - Two A-type proanthocyanidins from Prunus armeniaca roots. AB - Two A-type proanthocyanidins, ent-epiafzelechin-(4alpha-->8, 2alpha-->O-->7)-(-) epicatechin and its isomer, ent-epiafzelechin-(4alpha-->8', 2alpha-->O-->7') catechin, together with other known compounds, have been isolated from the roots of Prunus armeniaca. The concerted use of one- and two-dimensional NMR methods (1H-(1)H COSY, HMQC, HMBC and NOESY) allowed the identification of these compounds. PMID- 10844161 TI - Pycnogenol in chronic venous insufficiency. AB - Forty patients with chronic venous insufficiency (CVI) and varices of the legs were selected and double-blindly randomly assigned to a treatment with Pycnogenol (French maritime pine bark extract), 100 mg x 3/day or a placebo for 2 months, according to a double-blind experimental design. The effects of the treatment were evaluated by scoring the symptomatology with a semi-quantitative scale, and the venous blood flow by means of a hand-held Doppler ultrasound. The tolerability was evaluated by recording the adverse effects and by means of hematology and blood chemistry parameters, before and at the end of the treatment. Pycnogenol treatment induced a significant reduction in subcutaneous edema as well as heaviness and pain in the legs, on both after 30 and 60 days, the evaluation time periods. Approximately 60% of patients treated with Pycnogenol(R) experienced a complete disappearance of edema (the most rapidly disappearing symptom) and pain at the end of the treatment, while almost all the patients reported a reduction in leg heaviness which disappeared in approximately 33% of patients. These changes were statistically significant. No effect was observed in the placebo-treated subjects. No effect on the venous blood flow was observed in either of the experimental groups. PMID- 10844163 TI - Comparative studies of the immunomodulatory activity of Tinospora cordifolia and Tinospora sinensis. AB - The water and ethanol extracts of stems of Tinospora cordifolia and T. sinensis inhibit immunosuppression produced by cyclophosphamide. Ethanol extracts of stems of both the plants inhibit cyclophosphamide-induced anemia. The water extract of T. sinensis is found to be more potent than the other extracts. PMID- 10844164 TI - Triterpenoid p-aminobenzoates from the seeds of zucchini. AB - An investigation of the triterpenoid fraction of zucchini seeds afforded two novel multiflorane p-aminobenzoates, identified as 7-epi zucchini factor A and debenzoyl zucchini factor B. Multiflorane p-aminobenzoates could not be detected in zucchini sprouts, which contained bryonolic acid as the only multiflorane constituent. No compound of this type could be obtained from adult plant parts (roots, stems, leaves). PMID- 10844165 TI - Eudesmanoids from Sphaeranthus indicus. AB - Three new eudesmanoids have been isolated from Sphaeranthus indicus and their structures were established as 11alpha,13-dihydro-3alpha,7alpha-dihydroxy-4,5 epoxy-6beta,7-eudesmanolide, 11alpha,13-dihydro-7alpha-acetoxy-3beta-hydroxy 6beta,7-eudesm-4-enolide and 3-keto-beta-eudesmol by comparison of spectral data with those of other 7alpha-hydroxyeudesmanolides. PMID- 10844166 TI - Further sesquiterpenoids and phenolics from Taraxacum officinale. AB - Five germacrane- and guaiane-type sesquiterpene lactones, including two previously described taraxinic acid derivatives, were isolated from the roots of Taraxacum officinale, together with benzyl glucoside, dihydroconiferin, syringin and dihydrosyringin. The other three lactones were identified as 11beta, 13 dihydrolactucin, ixerin D and ainslioside. Moreover, the stereochemistry at C-11 in dihydrotaraxinic acid was assigned. PMID- 10844168 TI - Contribution to the knowledge of Rifian traditional medicine. II: Folk medicine in Ksar Lakbir district (NW Morocco). AB - An ethnobotanical survey of the medicinal plants used by the local population of the Ksar Lakbir district (NW Morocco) was conducted. One hundred and eighty-six species from 61 botanical families were recorded as well as their uses and modes of administration. Quantitative ethnopharmacological data (medicinal plant knowledge and use indices) were also evaluated and discussed. PMID- 10844167 TI - New flavonoids from Avicennia marina. AB - In addition to luteolin 7-O-methylether, chrysoeriol 7-O-glucoside and isorhamnetin 3-O-rutinoside, the methanol extract of the aerial parts of Avicennia marina yielded two new flavonoids identified as luteolin 7-O methylether 3'-O-beta-D-glucoside and its galactoside analogue 2. PMID- 10844169 TI - Molluscicidal activity of some Moroccan medicinal plants. AB - Among 14 plants of Moroccan folk medicine tested for molluscicidal activity, ethyl acetate extract from Origanum compactum and hexane extracts from both Chenopodium ambrosioides and Ruta chalepensis were the most active (LC(90)=2.00, 2.23 and 2.23 mg l(-1), respectively) against the schistosomiasis-transmitting snail Bulinus truncatus. PMID- 10844170 TI - Antibacterial activity of Enantia polycarpa bark. AB - Antimicrobial screening of Enantia polycarpa bark showed activity of the polar extracts against some microorganisms. PMID- 10844171 TI - Antifungal activity of some tetranortriterpenoids. AB - Natural tetranortriterpenoids such as cedrelone from Toona ciliata, azadiradione from Azadirachta indica, limonin, limonol and nomilinic acid from Citrus medica, along with some cedrelone derivatives were tested for their antifungal activity against Puccinia arachidis, a groundnut rust pathogen. Results show that cedrelone was the most effective in reducing rust pustule emergence. Replacement of functional groups or modification of the A or the B ring in cedrelone reduced the effectiveness indicating the importance of specific structural features for activity. PMID- 10844173 TI - Antibacterial activity of Dichrostachys cinerea. AB - The antibacterial activities of the chloroform, methanol and aqueous extracts of Dichrostachys cinerea fruits and leaves are reported. PMID- 10844172 TI - Antibacterial activity of xanthorrhizol from Curcuma xanthorrhiza against oral pathogens. AB - The antibacterial activity of xanthorrhizol, isolated from the methanol extract of Curcuma xanthorrhiza roots, was evaluated against oral microorganisms in comparison with chlorhexidine. PMID- 10844174 TI - Alkaloids from Balanites aegyptiaca. AB - High performance liquid chromatographic (HPLC) analysis of a dichloromethane extract of the stem-barks of Balanites aegyptiaca has yielded two known alkaloids, N-trans-feruloyltyramine (1) and N-cis-feruloyltyramine (2), and three common metabolites, vanillic acid, syringic acid and 3-hydroxy-1-(4-hydroxy-3 methoxyphenyl)-1-propanone. PMID- 10844175 TI - Chemical constituents from Asparagus dumosus. AB - The isolation and characterization of calonysterone (1), blechnoside B (2), 20 hydroxyecdysone and isovanillin from the whole plant of Asparagus dumosus are being reported for the first time from this source. PMID- 10844176 TI - Minor furocoumarins of Murraya koenigii. AB - Xanthotoxin, isobyakangelicol, phellopterin, gosferol, neobyakangelicol, byakangelicol, byakangelicin and isogosferol are reported as minor furocoumarins of Murraya koenigii seeds. PMID- 10844177 TI - Saponins from Chenopodium album. AB - The isolation and spectral data of three saponins from the roots of Chenopodium album L. are reported. One of them is a seco-glycoside analogous to compounds that were previously found in species belonging to Caryophyllales. PMID- 10844178 TI - Coumarinolignoids of Mallotus apelta. AB - Three coumarino-lignoids, aquillochin (1), cleomiscosin A and 5' demethylaquillochin have been isolated from Mallotus apelta. The structure of the new compound 3 was determined by spectroscopic techniques. PMID- 10844179 TI - Herbal medicinal products. (Their place in healthcare, scientific and regulatory agenda for the future) October 8th, 1999 Amsterdam. PMID- 10844180 TI - Preface PMID- 10844181 TI - New opportunities in animal breeding and production - an introductory remark. AB - New opportunities in animal breeding and production will be offered by methods for embryo production and the introduction of precise genetic changes in livestock species. A review of current procedures shows that they have been improved during the last 4 years, but still have significant limitations. In the light of experience over the past 50 years it seems likely that these methods will contribute to animal health and productivity, but at some unknown time in the future and in ways that cannot be predicted precisely. When looking to the future, delegates to this meeting should take pride in what science continues to contribute. PMID- 10844182 TI - Bioethics: limits to the interference with life. AB - The paper presents a review of various ethical considerations to which the application of modern biotechnology in breeding of domestic animals gives rise. The review is based on an automated literature search, covering papers and reports within agricultural bioethics published since 1992. The aim is to present the different points of view in a clear, unbiased manner. First the various concerns and viewpoints are presented. The concerns are divided into three main categories: animal welfare concerns, concerns about animal integrity and concerns relating to human health and environmental issues. Then follows a discussion of how to weigh potentially conflicting concerns against each other. The paper concludes with suggestions about how ethical issues should be handled in practice. PMID- 10844183 TI - Neuroendocrine regulation of gonadotropins in the male and the female. AB - For the past decade, neuroendocrinology, in general, and neuroendocrine regulation of reproduction, in particular, were strongly dominated by molecular genetics and molecular endocrinology. In very recent years, however, neuroendocrinology is taking back its place. Beyond doubt GnRH is the neuroendocrine signal for ovulation. But there are still many unexplored pathways within the 'black box' triggering and regulating this signal. Neuroendocrine control of reproduction starts very early in life, well before birth. Hypophyseal gonadotropin secretion is under hypothalamic control at around mid-gestation in the fetal sheep and the fetal pig. These two species could be considered as best studied farm animals considering neuroendocrinology. This minireview thus will give in the first part a short survey of developmental processes of some of the neuroendocrine systems in the pig and sheep. In the second part, the opioidergic and catecholaminergic control of gonadotropins in adults will be briefly discussed. The last part will focus on the new less known pathways mediating effects on hormones which regulate the reproductive functions. PMID- 10844184 TI - Activation of ribosomal RNA genes in preimplantation cattle and swine embryos. AB - Transcription of ribosomal RNA (rRNA) genes occurs in the nucleolus resulting in ribosome synthesis. In cattle and swine embryos, functional ribosome-synthesizing nucleoli become structurally recognizable towards the end of the fourth and third post-fertilization cell cycle, respectively. In cattle, a range of important nucleolar proteins become localized to the nucleolar anlage over several cell cycles and this localization is apparently completed towards the end of the fourth cell cycle. In swine, the localization of these proteins to the anlage is more synchronous and occurs towards the end of the third cell cycle and is apparently completed at the onset of the fourth. The rRNA gene activation and the associated nucleolus formation may be used as a marker for the activation of the embryonic genome in mammalian embryos and, thus, serve to evaluate the developmental potential of embryos originating from different embryo technological procedures. By this approach, we have demonstrated that in vitro produced porcine embryos display a lack of localization of nucleolar proteins to the nucleolar anlage as compared with in vivo developed counterparts. Similarly, bovine embryos produced by nuclear transfer from morulae display such deviations as compared with in vitro produced counterparts. Collectively, this information may help to explain the appearance of abnormalities seen in a certain proportion of offspring derived from in vitro produced embryos and after cloning. PMID- 10844185 TI - Selection of the dominant follicle in cattle and horses. AB - The nature of selection of the dominant follicle is reviewed by comparing research results between cattle and horses. In both species, emergence of a follicular wave is stimulated by an FSH surge. The surge reaches a peak by the time the follicles attain 4 mm in diameter in cattle and 13 mm in mares. In cattle, all of the growing follicles >/=5 mm contribute to the decline in FSH concentrations. However, the declining FSH concentrations are still needed by the growing follicles. Several days after the peak of the FSH surge and emergence of the wave, the two largest follicles reach means of 8.5 and 7.7 mm in cattle and 22 and 19 mm in horses. At this approximate time, the follicles begin to undergo deviation in follicle diameters, which is characterized by continued growth of the largest follicle to become the dominant follicle and reduced or terminated growth of the remaining follicles to become subordinate follicles. In both species, on average, the future dominant follicle emerges before the future largest subordinate follicle, and the two follicles grow in parallel until deviation. The difference in diameter between the two largest follicles at the beginning of deviation is equivalent in growth to approximately 8 h in cattle and 24 h in mares. Apparently, this is adequate time for the largest follicle to establish the deviation process before the second-largest follicle reaches a similar diameter. During this time, the largest follicle plays the primary role in further suppressing circulating FSH concentrations to below the requirements of the smaller follicles, which causes their regression. The follicle-produced FSH suppressants appear to be estradiol and inhibin. In addition to enhancing its FSH-suppressing ability, the largest follicle also develops the ability to utilize the reduced concentrations of FSH for its continued growth. It is therefore postulated that the essence of selection of a dominant follicle in these two species is a close two-way functional coupling between changing FSH concentrations and follicle growth and development. Elevated concentrations of circulating LH encompass deviation in both species and may play a role in continued growth of the largest follicle. It is not known if LH begins to be utilized by the largest follicle before, at, or after the beginning of diameter deviation. However, results of studies in mares suggested that LH does not influence growth of the dominant follicle until after the beginning of deviation. PMID- 10844186 TI - In vitro culture of bovine embryos in Menezo's B2 medium with or without coculture and serum: the normalcy of pregnancies and calves resulting from transferred embryos. AB - The present study was designed to test the efficacy, as assessed by blastocyst production, of culturing slaughterhouse-derived bovine oocytes in Menezo's B2 (B2) medium with or without serum and with or without buffalo rat cell (BRL) coculture. In addition, OPU-derived oocytes were cultured in B2-BRL coculture with or without serum for the first 72 h or TCM 199-BRL coculture with serum and the resulting embryos were transferred into recipients. Culture in B2 plus serum resulted in more blastocysts than B2 without serum, but both treatments produced far fewer blastocysts than did B2-BRL coculture. In addition, B2 with or without serum produced embryos of a lower stage of development, lower quality and fewer cells than did B2-BRL coculture. Serum restriction for the first 72 h of B2-BRL coculture did not result in a detectable difference in blastocyst production during 7 or 8 days of culture relative to coculture with no serum restriction. Embryos produced in B2-BRL coculture with serum restriction resulted in pregnancy rates, percentage of male fetuses, abortions, and congenital problems similar to those from B2 or TCM 199-BRL coculture with no serum restriction. PMID- 10844187 TI - Sexing mammalian sperm for production of offspring: the state-of-the-art. AB - Predetermination of sex in livestock offspring is in great demand and is of critical importance to providing for the most efficient production of the world's food supply. With the changes that have taken place in animal agriculture over the past generation the application of sex preselection to production systems becomes increasingly necessary. The current technology is based on the well-known difference in X- and Y-sperm in the amount of DNA present. The method has been validated on the basis of live births, laboratory reanalysis of sorted sperm for DNA content and embryo biopsy for sex determination. The technology incorporates modified flow cytometric sorting instrumentation to sort X- and Y-bearing sperm. Resulting populations of X or Y sperm can be used in conjunction with IVF in swine and in cattle for the production of sexed embryos to be transferred to eligible recipients for the duration of gestation. It can also be used for intratubal insemination and for deep-uterine and conventional insemination in cattle. This semipractical sexing method, though currently impractical for some production systems (where large numbers of sperm are required for fertilization) could be used to provide a more flexible progeny-producing option in many livestock operations. Improvements in the production rate of sexed sperm continue as new technology is developed. High-speed sorting is one of the newer technological advances and is being used in our laboratory to increase sorted sperm throughput. With our original technology we sorted 350,000 sperm/h. We now sort 6 million of each sex, under routine conditions. Sorting only the X population results in about 18 million sperm/h. Improvements in the technology will no doubt lead to much greater usage of sexed sperm, depending on the species involved. Insemination of lower sperm numbers in cattle has proven to be an effective means of utilizing the sexing technology. Solving the problems associated with inseminating low sperm numbers in the pig would be advantageous to the utilization of sexed sperm for some type of deep artificial insemination. Such a development would also enhance the economy of using lower sperm numbers with conventional artificial insemination (AI) and aid the swine industry worldwide. The use of sexed sperm for non-ordinary applications such as endangered species, laboratory animals, hobby or pet species is also of interest and will become a part of the move to be more reproductively efficient in the next millennium. Sexed sperm on demand over the next several years will provide livestock producers with many options in seeking to improve efficiency of production and improve quality of products to enhance consumer acceptability. PMID- 10844188 TI - To treat or not to treat: a proper use of hormones and antibiotics. AB - Hormones and antibiotics are important remedies in animal reproduction. Compared to other areas of application, hormones are probably more used than antibiotics. The quantities of hormones applied in cattle reproduction are largely dependent on whether these drugs are extensively used for pharmaceutical control of breeding or not. Diseased animals should be treated both from an animal welfare point of view and to restore their production capacity. The treatment should be based on an accurate diagnosis. Some of the treatment methods used in animal reproduction do not seem to be well documented. When using antibiotics, it should be known that an infectious agent is present which will be susceptible to therapy. The use of hormones and antibiotics to solve or mask managerial problems should be avoided. Ideally, fertility and health traits should be included in a breeding programme. Therefore, all diagnoses and treatments performed should be recorded and these data made available for breeding purposes. Manipulation of the breeding cycle by pharmaceutical means should not disturb the natural reproductive performance of animals being progeny tested. Animal health and fertility should be improved by selection and good management rather than by extensive use of hormones and antibiotics. Cases of inappropriate use of pharmaceutical preparations have created a general scepticism among people concerning the use of hormones and antibiotics in modern farming. Evidence of increasing resistance to antibiotics in bacteria infecting humans has focused on the role that anti-microbial drug use in food-producing animals plays in the emergence of resistant bacteria. There is also a concern about possible residues in animal products. Further, the consumers have a growing interest in animal health and animal welfare issues, and they have ethical concerns regarding the use of hormones and antibiotics, in particular, as performance enhancers. In Europe, the number of farmers growing organically cultivated foodstuffs is increasing, and according to the regulations for organic farming, the use of hormones and antibiotics is limited. Even though the proper use of hormones and antibiotics does not have any known negative effect on animal welfare or public health, the consumers' concerns have to be taken into account in livestock production. PMID- 10844190 TI - Embryonic mortality and embryo-pathogen interactions. AB - Embryonic mortality (EM) has a substantial impact on the fertility of domestic animals. Most of the embryonic losses occur during the first days after fertilization and during the process of implantation. Causes of EM can be divided into infectious and non-infectious categories. Primary attention has often been given to infectious agents but non-infectious causes probably account for 70% or more of the cases of embryonic death. Infection of the embryonic environment can be caused by specific and non-specific uterine pathogens. Specific uterine infections are caused by a number of viruses, bacteria and protozoa that enter the uterus by the haematogenous route or via the vagina. Non-specific pathogens are mainly bacteria that enter the uterus by ascending infection. Uterine pathogens may cause EM by changing the uterine environment (endometritis) or by a direct cytolytic effect on the embryo. Non-infectious causes of EM such as chromosomal aberrations, external factors (e.g., high ambient temperature and nutritional factors) and maternal factors (e.g., hormonal imbalances and age) are multifactorial and difficult to diagnose. PMID- 10844189 TI - Deteriorating trends in male reproduction: idiopathic or environmental? AB - Recent reports portend deterioration in male reproductive health in several human populations. Similar trends might exist in domestic animals, but data are not available because of the inherent nature of animal husbandry practices - culling of the reproductively inefficient food- and fiber-producing animals at an early age. Although the causes for this deterioration are unknown, a variety of endocrine-mimicking environmental pollutants have been implicated. Data for relevant laboratory animal models exposed to several classes of suspect chemicals indicate that a variety of chemicals ubiquitously present in the environment can disrupt normal reproductive phenomena in the male at exposure rates encountered in nature. Data are presented for occurrence of cryptorchidism, carcinoma in situ of the testis, acrosomal malformations, and impaired sexual function following in utero and/or postnatal exposures to pesticides (e.g., DDT and vinclozolin), high volume industrial chemicals (e.g., alkylphenols and phthalates), and commonly occurring organic and inorganic chemical contaminants in drinking water (e.g., chemical mixtures and water disinfection byproducts). These observations are discussed in the context of similar, so-called idiopathic conditions encountered in stallions. PMID- 10844191 TI - The bovine placenta before and after birth: placental development and function in health and disease. AB - This paper reviews bovine placental development, anatomy (microscopic and gross), nomenclature and classification. The paper focuses on the biology of those specialized cells that arise from the outermost layer of very early embryos, the trophoblast cells, and on placental macrophages, cells that play a key role in fetal/placental defense. Data is presented from an immunohistochemical quantitative study that characterizes the ontogeny of placental macrophages using placental tissues from 21 cows (sampled from 4 months of pregnancy through the post partum period). Understanding of bovine placental development is essential for veterinarians, pathologists, diagnosticians and researchers. Lesions of diagnostic significance can be recognized for many economically important infectious abortifacient diseases, and there is growing evidence that pregnancy failure of cloned calves is due in part to unexplained placental failure. Placentology and placental pathology are becoming of increasing importance. PMID- 10844192 TI - Reproductive biotechnologies: current status in porcine reproduction. AB - During the past decade, considerable attention has been directed toward the development of reproductive technologies for both research purposes and for more controlled swine reproduction. Artificial insemination is an example of a technology that has continued to be expanded from early use in European countries to the USA and Canada where it is now estimated that a majority of the sows bred are artificially inseminated. In addition, several significant technological advancements have been made in the genetic modification of swine and interest has been generated in the possible use of swine as donors of specific tissues and of organs for the improvement of human health. At the same time, the systems for production of swine for human food continue to undergo major changes including, in some countries, the consolidation of swine into large, integrated units. These swine operations are very receptive to the use of technologies to reduce labor costs as well as a basis for increased production efficiency. Therefore, the combined interest in swine reproductive technologies by both the medical field and the swine industry creates an increased effort for the development of new technologies as well as for the implementation of existing ones. One of the more rapid technological advancements this decade has been the progress in in vitro production (IVP) of swine embryos. Major advancements have been made on the development of procedures for production of large numbers of embryos from oocytes collected at slaughter houses which are then matured (IVM) and fertilized (IVF) in the laboratory. Success in IVP has stimulated increased research in other areas that can be enhanced by the availability of embryos without a requirement for surgical collection from gilts or sows. One example is the combined use of IVF, gender-sorted sperm cells, and embryo transfer to produce offspring of a predicted sex. In a related area, instrumentation for non-surgical embryo transfer has recently been developed that results in significant improvement in this technology. Similar achievements have been gained in cryopreservation of embryos by vitrification. These developments will be reviewed with emphasis on the in vitro production of embryos from immature oocytes. PMID- 10844193 TI - Factors effecting reproduction in the pig: seasonal effects and restricted feeding of the pregnant gilt and sow. AB - Recent advances in research on seasonal infertility are discussed with a special focus on implications of the generally recommended restricted post-mating feeding strategy of the early pregnant gilt and sow for the physiology of seasonal infertility. The endocrinological basis of seasonal breeding of the wild and domestic pig is being clarified: as in other seasonal breeders, melatonin is relaying photoperiodic information about season to the pituitary-gonadal axis. Earlier confusion on this matter appears to have been caused by a lack of specificity of the melatonin assays employed. Group housing of the pregnant sow is becoming a common practice and, as an important environmental risk factor for seasonal infertility, may lead to an increase in the incidence of seasonal infertility in the future. After an initial progesterone-mediated beneficial effect on embryonic survival, a restricted post-mating feeding strategy may have a negative effect on maintenance of early pregnancy in the gilt and sow in the summer-autumn period. The endocrinological mechanism of seasonal disruption of pregnancy is yet to be determined. However, it is proposed that LH is reduced in the summer-autumn period and this reduction is amplified by the commonly applied restricted post-mating feeding strategy. These changes in LH secretion, although not as such inducing CL regression, may exert a progesterone-mediated detrimental effect on the capability of embryos to produce adequate embryonic signaling. This may lead to a seasonal disruption of pregnancy and a return to oestrus 25-30 days after mating. PMID- 10844194 TI - Influence of the nutritional status on ovarian development in female pigs. AB - In female pigs, undernutrition may influence growth of antral follicles from various size classes, decrease ovulation rate, delay puberty and return to oestrus after weaning. It could also affect the oocyte maturation and hence the number of viable embryos per litter. Inhibition of the gonadotrophin release due to undernutrition is presumably involved in these phenomena. Presence of receptors, as well as in vitro and in vivo studies suggest that insulin and hormones from the somatotrophic axis are able to alter folliculogenesis directly at the ovarian level. They should act as hormones controlling nutrition, proliferation, growth and differentiation of the cells and/or as amplifiers of the action of gonadotrophins. Information are needed to determine whether their availability at the ovarian level may become insufficient or excessive in case of nutritional deficit. Increase in plasma concentrations of progesterone due to lower hepatic metabolic rate in underfed females probably contributes to inhibit folliculogenesis. PMID- 10844195 TI - The effect of porcine parvovirus and porcine reproductive and respiratory syndrome virus on porcine reproductive performance. AB - From a worldwide perspective, porcine parvovirus (PPV) and porcine reproductive and respiratory syndrome virus (PRRSV) are the most common viral causes of porcine reproductive failure. A typical epidemic of PPV-induced reproductive failure is presented as an increased number of mummified fetuses and sometimes, entire litters are mummified. If infection with PPV is very early in gestation, the number of liveborn pigs may be further reduced as a result of embryonic death and resorption. During the acute stage of infection gilts and sows have few, if any, clinical signs, and it is unlikely that PPV is ever the direct cause of abortion. In contrast, a typical epidemic of PRRSV-induced reproductive failure is presented as a broader spectrum of clinical features including abortions, late term dead fetuses, stillborn pigs, and weakborn pigs. In the later stages of an epidemic, there may also be an increase in the number of mummified fetuses, but their prevalence is likely to be far less than during an epidemic of PPV-induced reproductive failure. During the acute stage of infection with PRRSV, gilts and sows may have few, if any, clinical signs, or they may be severely affected and even die. This difference largely reflects the relative virulence of the strain of PRRSV causing the epidemic. A timely and reliable laboratory diagnosis of either disease can be made when appropriate tests are performed with appropriate samples. Vaccines are available for prevention of both diseases. PMID- 10844196 TI - Reproductive behavior of stallions and mares: comparison of free-running and domestic in-hand breeding. AB - Based on observational studies comparing social organization and reproductive behavior of equids breeding under feral, semi-feral, and domestic conditions, a series of comparisons and findings on reproductive physiology and behavior are presented. Simple changes in management and handling of mares and stallions that can improve reproductive efficiency and fertility or that can overcome specific breeding problems are discussed. PMID- 10844197 TI - Post-breeding endometritis in the mare. AB - Post-breeding endometritis is a major cause of subfertility in the mare. Endometritis is a normal event in the immediate period after mating, but the presence of ultransonographically visible uterine fluid more than 12 h later is thought to be evidence of uterine pathology. In mares that are free of venerally transmitted endometritis, treatment is aimed at removing the intraluminal fluid. If the endometritis persists past day 5, when the embryo enters the uterine lumen, the cytotoxic environment will not be compatible with pregnancy. Reproductive anatomy, defective myometrial contractility, lowered immune defences, overproduction of mucus, inadequate lymphatic drainage, or a combination of these factors will predispose the mare to post-breeding endometritis. PMID- 10844198 TI - Twin reduction in the mare: current options. AB - Twin pregnancy in the mare causes economic loss. Early transrectal ultrasonographic detection of twins and manual crush of one embryonic vesicle is the method of choice for managing equine twins (90% success rate). Transcutaneous and transvaginal ultrasound-guided twin reduction techniques are described for twin pregnancies that advance beyond 25 days of gestation. Reported success rates for the ultrasound-guided procedures are 50% and 20%, respectively. PMID- 10844199 TI - Seasonality in mares. AB - In this review, we have attempted to summarize, based on recent data obtained in our laboratory and elsewhere, our current understanding of the regulatory mechanisms of seasonality and discuss the implications with regard to treatment strategies to advance the onset of cyclic reproductive activity in the early spring. PMID- 10844200 TI - In vitro culture and embryo metabolism of cattle and sheep embryos - a decade of achievement. AB - At the beginning of the 1990s, co-culture of cattle and sheep embryos was the most favoured method to support embryo development, but the use of this system has hampered progress in raising the efficiency of embryo production. Furthermore, little was known of the requirements of embryos and the biochemistry of early embryo development. As the decade progressed, energy metabolism studies improved our understanding of the energy substrate requirements for embryo development. Furthermore, an appreciation of the reproductive tract environment increased. This resulted in more "defined" systems, which have evolved further in the development of "sequential" media systems, where components change in accordance to the needs of the embryo. Nevertheless, wholly defined systems, such as the replacement of albumin with PVA, are less able to support similar levels of development as protein-containing medium, and the resulting embryos are metabolically compromised. This highlights the nutritive role of albumin. One area in which much work has been conducted, but yet no unifying theory has emerged, is that of the interactive roles of growth factors (including autocrine/paracrine), cytokines and extra-cellular matrix molecules in the development of a viable embryo. A new concept is that of regulation of energy metabolism. Compounds such as ethylenediamine tetraacetic acid (EDTA), NaN(3) and 2,4-dinitrophenol have been shown to increase embryo development and quality of resulting embryos. This demonstrates that the process of ATP production is a key regulator of in vitro embryo development. PMID- 10844201 TI - Transgenic livestock: premises and promises. AB - Microinjection of DNA constructs into pronuclei of zygotes has been the method of choice for the generation of transgenic livestock. However, this procedure is characterized by low efficiency (1-4% transgenic offspring), random integration and variable expression of the transgene as well as a considerable proportion of mosaicism. Furthermore, it is extremely time consuming and costly. As a consequence, commercial application has focused on the production of recombinant proteins in the mammary gland of transgenic animals and xenotransplantation, e.g. the use of porcine organs in human organ transplantation. In addition, transgenic pigs carrying a modified porcine growth hormone (hMt-pGH) construct show significant improvements in economically important traits without adverse side effects of a GH overproduction. Improvements of transgenic technology will likely come from the generation of appropriate cell lines suitable for transfection or even homologous recombination and their subsequent use in nuclear transfer. Additionally, in the mouse a number of sophisticated molecular tools have been developed that allow precise modifications of the genome. These include the application of artificial chromosomes from yeast (YAC) or bacteria (BAC) for position-independent and copy-number-dependent expression of a transgene, the Tet system (tetracycline inducible) for a tight temporal control of transgene expression, as well as conditional mutagenesis by applying site-specific DNA recombinases (e.g. Cre, FLP). The successful adaptation of these molecular tools to livestock will enable the fulfillment of many of the promises originally thought to be achievable when transgenic livestock were first reported. PMID- 10844202 TI - Receptor blockers - general aspects with respect to their use in domestic animal reproduction. AB - Receptor blockers compete with the respective agonist for binding to a given receptor without inducing complete signal transduction. In recent years, major interest has focused on sex-steroid hormone receptor blockers (antagonists). Indications have been obtained that inadequate changes in receptor conformation and subsequent failure of transcriptional activation are major events preventing hormonal activity. However, various subtypes and variants of receptors and receptor mutations have also been identified. Expression of antihormonal effects may vary depending on the type of receptor the blocker is bound to. Hence, receptor blockers may also have an inherent agonistic activity. Aglepristone is the first antiprogestin registered for veterinary use with the indication "interruption or prevention of pregnancy"; similarly, these types of compounds were successfully used for induction of parturition in the dog and cat and for conservative treatment of pyometra in the dog. Moreover, application of antiprogestins has clearly demonstrated the role of progesterone as a major factor controlling overt pseudopregnancy in dogs. With respect to farm animals, parturition was induced in cows without an increased incidence of retained fetal membranes. Other than antiprogestins, antioestrogens and antiandrogens are still in a more experimental phase. In particular for use in humans, high-affinity blockers binding to the oxytocin/vasopressin receptor are in development; they exert distinct tocolytic activities. Also, the release of GnRH can be inhibited by respective antagonists; however, their use in reproduction is still hampered by the high dose requirement and the side effects observed. PMID- 10844203 TI - Gonadal steroid receptors in the regulation of GnRH secretion in farm animals. AB - The sites of action and mechanisms by which gonadal steroids regulate gonadotrophin-releasing hormone (GnRH) in domestic animals remain largely unknown. This review summarises information gained from sheep regarding the distribution of the gonadal steroid receptors in the brain, the neurochemical identity and the projections of these steroid receptor-containing neurones. The cells in the hypothalamus that contain each of the gonadal steroid receptors (oestrogen receptor alpha (ERalpha), oestrogen receptor beta (ERbeta), progesterone receptor (PR) and androgen receptor (AR)) show a remarkably similar distribution, although the PR and AR-containing cells are less widespread than oestrogen receptors (ERs). There is considerable overlap in the distribution of ERalpha- and ERbeta-containing cells but also some unique sites for each subtype. This suggests differential regulation of the actions of oestrogen. There appears to be little sexual dimorphism in the distribution of the gonadal steroid receptors in the hypothalamus, with the notable exception of the ventromedial nucleus where females appear to have greater numbers of both ERalpha- and ERbeta containing cells. Neuronal tracing studies have identified projections of some of the ERalpha-containing cells to sites that may allow interaction with the GnRH system. The receptor mapping, neuronal tracing and microimplantation studies suggest that the ventromedial nucleus is likely to be a key hypothalamic nucleus in the steroid regulation of GnRH secretion in sheep. PMID- 10844204 TI - Can we use in vitro fertilization tests to predict semen fertility? AB - This presentation deals with assays based on in vitro fertilization (IVF) and related techniques such as zona pellucida (ZP) binding assays and oocyte penetration tests. These types of assays have been developed for several species of domestic animals. A description of the assays and how they have been performed in domestic animals, as well as data on the correlation between the results of assays and actual in vivo fertility are presented. Used either as single tests or in combination with other tests, this type of assay can provide valuable information about a semen donor, an insemination dose or a method of semen preservation. PMID- 10844205 TI - A glimpse at sperm function in vivo: sperm transport and epithelial interaction in the female reproductive tract. AB - The process of sperm transport in the female reproductive tract is more than simply a migration of spermatozoa from the site of insemination to the site of fertilization. Rather, it is a complex and dynamic continuum that encompasses phases of sperm distribution within the tract, the accumulation of spermatozoa in reservoirs, the modulation of sperm physiology and acquisition of fertilization competence, the ascent of competent spermatozoa to the site of fertilization, and the elimination of the non-fertilizing sperm population. The dynamic interactions that occur between functional spermatozoa and the luminal fluids and epithelial surfaces of the female genital tract during transit and storage enhance sperm survival and regulate sperm function in the female. The universal nature of this interaction highlights it as a key component of the sperm transport process. PMID- 10844206 TI - Conservation of sperms: current status and new trends. AB - Preserving genetic resources for next millennium is of great importance, whereas the major contribution of conserving sperms has the potential in many applications such as agriculture, biotechnology, species conservation and clinical medicine. Two major systems (liquid and frozen) of storage technologies have been developed for sperm conservation until the 20th century. The combinations of storage temperature, the cooling rate, chemical composition of extender, cryoprotectant concentration and the hygienic control are the key factors that affect the life span of spermatozoa. In the past decades, a slow progress have been made in improvement of storage technology, however, the recent advancement in reproduction technology and well understanding of the reproductive physiology have opened the door to a new era in conservation of sperm. This paper focuses on current sperm conservation systems as well as alternative strategies that would be effective for preserving genetic resources in future. PMID- 10844207 TI - Vitrification of the oocytes and embryos of domestic animals. AB - After the first successful application of vitrification for embryo cryopreservation 15 years ago, a rapid application of the method in domestic animal embryology was presumed. However, although the advantages of vitrification (simplicity, cost efficiency, speed of the procedure) were widely acknowledged, its use has been mainly restricted to experimental studies. For commercial embryo transfer purposes, the traditional slow-rate or equilibrium freezing has been used. This review attempts to explain the reasons for this phenomenon and discusses the theoretical and practical differences between the two technologies as well as their commercial prospects. Recent developments that improve the efficiency of vitrification and applications to other reproductive technologies are also summarized. These advances may result in considerable advantage and could lead to widespread application of vitrification in certain areas of domestic animal embryology. PMID- 10844208 TI - Immunocontrol in dogs. AB - Population control in dogs and cats is an important goal for many groups. Control measures over the years has included surgery, hormonal therapy and more recently immunological control. The current presentation discusses dog population control with an emphasis on immunologic control. Specifically, vaccination with purified zona pellucida (ZP) glycoproteins leads initially to immunocontraception and then to the profound and irreversible changes of immunosterilization. The preliminary studies are extremely encouraging on developing a vaccine for lasting canine population control. PMID- 10844209 TI - Current state in biotechnology in canine and feline reproduction. AB - Biotechnology has proceeded much further in cats than in canines, although the pregnancy rate after in vitro maturation (IVM), IVC and embryo transfer (ET) is still relatively low. The use of AI with frozen-thawed semen as a breeding tool to overcome breeding incompatibility or to preserve male genetic material has been limited in felines in contrast to the situation in domestic dogs and foxes. In many research scenarios and endangered felid species programs, the in vitro production of feline embryos with subsequent transfer has complemented the use of AI. Improvement of IVM, in vitro fertilization (IVF) and embryo culture coupled with ovarian tissue grafting, cryobanking of follicles, oocytes, semen, or embryos, with subsequent ET into surrogate females, may render this technology feasible for use in endangered wild felids. In canines, reliable systems for in vitro production of embryos, embryo cryopreservation and transfer are yet to be developed. The refinement of invasive fertilization techniques, such as intracytoplasmic sperm injection (ICSI), may eventually provide a tool for removal of recipient oocyte nuclei and transfer of selected embryonic or somatic cell donor nuclei into domestic cat ooplasm, thereby providing a tool for genetic modification, or for preservation of valuable genetic material. PMID- 10844210 TI - Piecing together the puzzle of carnivore reproduction. AB - Recent advances in feline and canine reproductive studies demonstrate how methodically piecing this information together is beginning to reap rewards for wildlife conservation programs. Non-invasive endocrinology can be used to monitor female reproductive function, time con-specific introductions or AI, and diagnose pregnancy. Sperm morphology characteristics and cell membrane function may be genetically inherited and differ between genetically diverse and inbred species/populations in felids. It is not clear if the same is true for the endangered red wolf. While standards exist for freezing feline and canine sperm, new information using fluorescent staining and zona penetration assays (ZPA) indicates that significant damage can occur during pre-freeze cooling, and may also be related to a species' genetic diversity. Posthumous gamete salvage from genetically valuable animals not only provides a means to study sperm and oocyte physiology but also to assist with genetic management of populations. Using the knowledge gained, IVM/IVF and ICSI have been successful in the domestic cat and AI has resulted in offspring in numerous non-domestic felids. However, understanding the processes of IVM/IVF is still not well understood in canids. New information reveals that sperm and the cumulus cells may be integral to oocyte maturation and that canine epididymal sperm are not capable of undergoing fertilization. The acquisition of knowledge and application of biotechnologies lags behind for non-domestic canid conservation programs. PMID- 10844211 TI - Prostatic disorders in the dog. AB - Common canine prostatic disorders include benign prostatic hypertrophy (BPH), prostatitis, prostatic cysts and prostatic adenocarcinoma. BPH is a spontaneous and age-related disorder of intact male dogs, which occurs in more than 80% male dogs over 5 years of age, and which is associated with clinical signs of sanguinous prostatic fluid, constipation and dysuria. BPH signs respond to castration or to finasteride treatment (0.1-0.5 mg/kg per os once daily), as finasteride inhibits conversion of testosterone to dihydrotestosterone, causing prostatic involution via apoptosis. BPH often occurs concurrently with prostatic infection, abscessation, cysts and neoplasia in the intact dog, and finasteride induced prostatic involution may be beneficial in treatment of all of these conditions except neoplasia. Two studies suggest that risk of prostatic adenocarcinoma is increased in neutered, compared to intact male dogs. Although canine prostatic neoplasia, unlike human prostatic neoplasia, usually does not respond to androgen deprivation, recent reports of prostatic intraepithelial neoplasia (PIN) in a high percentage of older male dogs, with and without prostatic adenocarcinoma, suggests that PIN may be a precursor to adenocarcinoma in the dog as it is believed to be in man. PMID- 10844212 TI - Neosporosis in cattle. AB - During the past decade, Neospora caninum infection has emerged as an important reproductive disease in cattle throughout the world. Abortion, occurring during the middle of gestation, is the primary clinical sign of the infection in cattle. Surveys in several countries from three continents have identified N. caninum infection as the major diagnosed cause of bovine abortion. Both endemic and epidemic patterns of abortion may occur in herds. An important feature of this disease is that the protozoan parasite is maintained in cattle as a chronic infection which can be passed on to the fetus during pregnancy. Two methods for the transmission of the infection in cattle have been proposed and are the subject of current investigations. Horizontal transmission utilizes a two-host life cycle whereby the cow is infected from ingestion of coccidial oocyst stages shed by the definitive host. Experimental infections have confirmed that the dog is a definitive host for the parasite. There is epidemiological evidence that the dog has a role in the prevalence of the infection but, as yet, no confirmation that the dog is the source for natural infections in cattle. Vertical transplacental transmission of the infection is an important route of infection in many herds. Vertical transmission occurs because fetal infection frequently does not result in abortion but rather the fetus survives to be a persistently infected animal. A heifer calf that is born congenitally infected is capable of transmitting the infection to the next generation when she becomes pregnant, thus maintaining the infection in the herd. The clinical outcome of transplacental fetal infection with N. caninum is likely determined by maternal and fetal immune responses which involve humoral, and most importantly, cell-mediated immune factors. The diagnosis of the infection is assisted through histopathology and immunohistochemical examination of aborted fetuses and serologic testing of cattle for evidence of infection. Several types of serologic tests, based on the use of culture-derived organisms or recombinant N. caninum antigens are available. There are no proven control methods for the prevention or treatment of neosporosis. Suggested control measures focus on programs to reduce the number of congenitally infected animals retained in the herd and to minimize the opportunity for postnatal transmission from the environment. PMID- 10844213 TI - Reproductive responses of cattle to GnRH agonists. AB - The response in cattle to treatment with gonadotrophin releasing hormone (GnRH) agonist includes downregulation of GnRH receptors on gonadotrophe cells, desensitisation of the anterior pituitary gland to endogenous GnRH, and the abolition of pulsatile release of LH. In bulls, a tonic pattern of LH release is associated with increased secretion of testosterone, which persists for the duration of treatment with GnRH agonist. The mechanism for this response in bulls has not been elucidated, but clearly pulsatile release of LH is not required to stimulate the synthesis of steroidogenic enzymes that sustain elevated secretion of testosterone. In heifers, desensitisation to endogenous GnRH prevents the occurrence of the pre-ovulatory surge release of LH, thus blocking ovulation. The latter provided the opportunity to evaluate the potential of a GnRH agonist bioimplant to control fertility in heifers under extensive management. Bioimplants that contained graded amounts of GnRH agonist prevented pregnancies in heifers for periods of 3 to 12 months. Zebu crossbred heifers treated with GnRH agonist from 14 to 23 months of age failed to conceive, but showed normal conception patterns when introduced into mating herds at around 26 months of age. After treatment with GnRH agonist for 4 to 6 weeks, ovarian follicular growth in heifers is restricted to relatively small (2-4 mm) antral follicles. Suppressed follicular growth in heifers treated long-term with GnRH agonist is due to a lack of gonadotrophin support, rather than a direct action of agonist at the ovaries. This was demonstrated by the ability to induce apparently normal follicular growth and ovulation by acute treatment with FSH for 4 days, followed by an injection of LH, in heifers that had been exposed to GnRH agonist for around 6 months, and which had only small (2-4 mm) antral follicles at the start of FSH treatment. GnRH agonist bioimplants have been incorporated into new multiple ovulation and embryo transfer protocols that allow control of the time of ovulation subsequent to superstimulation of ovarian follicular growth with FSH. In these protocols, the endogenous surge release of LH is blocked by treatment with agonist and ovulation is timed by injection of exogenous LH, allowing fixed time AI. It can be concluded from recent studies that GnRH agonist bioimplants have considerable potential for both pro-fertility and anti-fertility applications in cattle. It is likely that commercial bioimplants will be available within the next 3 to 5 years. PMID- 10844214 TI - Robot milking and effect on reproduction in dairy cows: a preliminary study. AB - The aim of this study was to analyse whether automatic milking has an effect on reproduction in dairy cows. Probably the most important consequence of automatic milking is an increased milking frequency. More frequent milking means more production and thereby an extra threat for the energy balance during the first months of lactation. A deep negative energy balance (NEB) is often followed by negative effects on fertility. However, tested under experimental circumstances, automatic milking with a frequency not higher than 3 times/day and with an individual management and feeding system has benefits for production and very little negative effects on reproduction. Experimental farms with automatic milking systems have fertility figures that do not differ from experimental farms with conventional milking systems. Additional and more data of automatic milking under farm conditions has to be collected and evaluated before the effect on fertility can be assessed with more certainty. PMID- 10844215 TI - Nutritional interactions with reproductive performance in dairy cattle. AB - Increased capability for milk production has been associated with a decline in fertility of lactating cows. Nutritional requirements increase rapidly with milk production after calving and result in negative energy balance (NEBAL). NEBAL delays the time of first ovulation through inhibition of LH pulse frequency and low levels of blood glucose, insulin and insulin-like growth factor-I (IGF-I) that collectively restrain estrogen production by dominant follicles. Up regulation of LH pulses and peripheral IGF-I in association with the NEBAL nadir facilitates ovulation. NEBAL reduces serum progesterone concentrations and fertility. Diets high in crude protein support high milk yield, but are also associated with lower reproductive performance. High protein can result in elevated plasma urea concentrations that affect the uterine environment and fertility. Nutritional interactions resulting in poor fertility of high producing dairy cows include the antecedent effects of NEBAL and effects of high dietary protein. PMID- 10844216 TI - Immunization against GnRH in male species (comparative aspects). AB - Active immunization against gonadotropin-releasing hormone (GnRH) was recognized in the 1970s as a potential means by which the reproductive system of mammals might be shut down for various practical and clinical reasons. Numerous studies in males have been performed since that time to determine the applicability of the technique as an alternative to surgical removal of the testes. Reasons for such immunocastration include improvement of meat and carcass characteristics for cattle, sheep, goats, and swine; improvement in feed efficiency relative to castrates in those same species; reduction in male aggressive behavior; reduction in male-associated odors in goats and swine; and fertility neutralization in pet species. Although application as a fertility control agent in men is unlikely, there is renewed interest in active immunization against GnRH as a means of treating prostate cancers and related steroid-dependent pathologies. PMID- 10844217 TI - Efficiency of spermatogenesis: a comparative approach. AB - Efficiency of spermatogenesis is the estimated number of spermatozoa produced per day per gram of testicular parenchyma. Spermatogenesis is the process of cell division and cell differentiation by which spermatozoa are produced in testes. Efficiency of spermatogenesis is influenced by species differences in the numerical density of germ cell nuclei and in the life span of these cells. Activities of spermatogonia, spermatocytes, and spermatids partition spermatogenesis into three major divisions (spermatocytogenesis, meiosis, and spermiogenesis, respectively). Spermatocytogenesis involves mitotic germ cell division to produce stem cells and primary spermatocytes. Meiosis involves duplication of chromosomes, exchange of genetic material, and two cell divisions that reduce the chromosome number and yield four spermatids. In spermiogenesis, spherical spermatids differentiate into mature spermatids which are released in the lumen of seminiferous tubules as spermatozoa. Spermatogenesis and germ cell degeneration can be quantified from numbers of germ cells in various developmental steps throughout spermatogenesis. Germ cell degeneration occurs throughout spermatogenesis; however, the greatest impact occurs during spermatocytogenesis and meiosis. There are species and seasonal influences on the developmental steps in spermatogenesis at which germ cell degeneration occurs. Number of Sertoli cells, amount of smooth endoplasmic reticulum of Leydig cells, and the number of missing generations of germ cells within the spermatogenic stage of the cycle influence efficiency of spermatogenesis. Efficiency of spermatogenesis is influenced to the amount of germ cell degeneration, pubertal development, season of the year, and aging of humans and animals. PMID- 10844218 TI - The causes of reduced fertility with cryopreserved semen. AB - Cryopreserved mammalian semen is generally acknowledged to have an impaired fertility by comparison with fresh semen. The reduction arises from both a lower viability post-thaw and sublethal dysfunction in a proportion of the surviving subpopulation. The reasons for the loss of fertility are various. In this paper, factors affecting the proportion of survivors (e.g., cold shock susceptibility, cooling rate, diluent composition and osmotic stress) and factors influencing functional status of survivors (e.g., membrane stability, oxidative damage, membrane receptor integrity, nuclear structure) are briefly reviewed. The possible effects of cryopreservation on the role of spermatozoa in the early stages of embryogenesis are considered. In the light of this review, indications for new approaches for improving the performance of cryopreserved semen are offered. PMID- 10844219 TI - Techniques for evaluating selected reproductive disorders of stallions. AB - Numerous techniques may be used for evaluation of the different reproductive disorders of the stallion. Approaches may vary from real-time ultrasonography and biopsy for evaluating testicular tumors to use of special assays for evaluating sperm or plasma for presence of antisperm antibodies. This communication addresses techniques used to evaluate five relatively uncommon, but perplexing, disorders of breeding stallions: (1) seminal vesiculitis, (2) hemospermia associated with idiopathic urethral defects, (3) acrosomal dysfunction, (4) abnormal spermatozoal chromatin, and (5) azoospermia. PMID- 10844220 TI - Effect of an inverse subtropical (19 degrees 13'N) photoperiod on ovarian activity, melatonin and prolactin secretion in Pelibuey ewes. AB - Twenty-one Pelibuey ewes were used from December 21, 1996 to December 21, 1998. Fourteen of them had never been exposed to artificial photoperiod, and they were maintained on natural photoperiod until March 21, 1997, when they were assigned to natural photoperiod (control group, n=8) or to inverse photoperiod (n=6). The other seven animals had been kept on a long photoperiod (16L:8D) from October 21, 1996 to December 21, 1996, when they entered the present study and were subjected to a gradual decrease in photoperiod, so that they reached an equinox photoperiod (12L:12 D) on March 21, 1997. At that time, they were assigned to natural photoperiod (n=3) or to inverse photoperiod (n=4). Blood samples for progesterone determination were taken twice a week from all the animals. During the second year of the study, prolactin was measured in the samples from five animals in inverse photoperiod and from five control ewes. Hourly samples were obtained to determine the 24-h melatonin profile of five animals from each group on September 21, 1997, December 21, 1997, March 21, 1997, and June 21, 1997. Exposure to inverse photoperiod resulted in a gradual shift on the annual reproductive cycle, so that the second ovulatory season was advanced by 5 months in the ewes kept on inverse photoperiod as compared to the control ewes (P<0.05). There were wide variations in the dates for the onset and the end of the ovulatory season within the inverse photoperiod groups, and three animals in this groups maintained ovulatory activity for at least 18 consecutive months. The duration of melatonin secretion was directly related to the length of the dark period (P<0.05), and this response was not affected by the calendar date. Prolactin concentrations were directly related to daylength, however, they were also affected by calendar date, being lower in the inverse group as compared to the corresponding time of the annual photoperiodic cycle of ewes on natural photoperiod. It is concluded that reproductive activity, melatonin secretion and prolactin secretion of Pelibuey ewes respond to the small variations in photoperiod that are present at 19 degrees 13'N, and that under natural conditions, photoperiod appears to be the main regulator of ovarian activity at this latitude. However, other factors such as temperature or humidity may act as modulators, and their relative importance could increase at more equatorial latitudes. PMID- 10844221 TI - Pig reproduction in South East Asia. AB - Asia with over half the population in the world has a thriving agriculture, a large part of which is devoted to pig production. The efficiency of production varies from country to country and is largely the legacy of whether the country relies on backyard farms using native breeds or intensive commercial farms using genetically improved breeds from Europe and North America. Reproductive efficiency is not as advanced as in Europe and this is the result of climatic conditions, which are hot and humid, the presence of most pig diseases, substandard management, the difficulty of obtaining quality feeds and appropriate delivery systems and the reliance on native breeds which are far less prolific than European ones. Improvements can only occur if investments are made in housing, cooling systems, disease control, management, quality feeds and better genetics. PMID- 10844223 TI - Micronutrients and reproduction in farm animals. AB - Reproductive well-being and performance of farm animals is largely dependent on their nutritional status, which is often less than optimum in developing tropical countries. More often than not, they are malnourished, particularly with regards to micronutrients. Evidence was presented to show that because these micronutrients are involved in such functions as intracellular detoxification of free radicals, synthesis of reproductive steroids and other hormones, carbohydrate and protein and nucleic acid metabolism, their deficiencies and/or excesses may impair spermatogenesis and libido in the male, fertility, embryonic development and survival, post-partum recovery activities, milk production and offspring development and survival. A plea was made for intensified research efforts, farmer education and quality control of vitamin-mineral pre-mixes, in order to improve micronutrient nutrition, and, consequently, the reproductive performance and overall productivity of farm animals in developing tropical countries. PMID- 10844222 TI - Impaired reproduction in heat-stressed cattle: basic and applied aspects. AB - Summer heat stress (HS) is a major contributing factor in low fertility in lactating dairy cows in hot environments. Although modern cooling systems are used in dairy farms, fertility remains low. This review summarizes the ways in which the functioning of various parts of the reproductive system of cows exposed to HS is impaired. The dominance of the large follicle is suppressed during HS, and the steroidogenic capacity of theca and granulosa cells is compromised. Progesterone secretion by luteal cells is lowered during summer, and in cows subjected to chronic HS, this is also reflected in lower plasma progesterone concentration. HS has been reported to lower plasma concentration of LH and to increase that of FSH; the latter was associated with a drastic reduction in plasma concentration of inhibin. HS impairs oocyte quality and embryo development, and increases embryo mortality. High temperatures compromise endometrial function and alter its secretory activity, which may lead to termination of pregnancy. In addition to the immediate effects, delayed effects of HS have been detected as well. Among them, altered follicular dynamics, suppressed production of follicular steroids, and low quality of oocytes and developed embryos. These may explain the low fertility of cattle during the cool autumn months. Hormonal treatments improve low summer fertility to some extent but not sufficiently for it to equal winter fertility. A limiting factor is the inability of the high-yielding dairy cow to maintain normothermia. A hormonal manipulation protocol, which induces timed insemination, has been found to improve pregnancy rate and to reduce the number of days open during the summer. PMID- 10844224 TI - Reproduction in female reindeer. AB - Reindeer are either wild or kept under very extensive farming systems. They are seasonal breeders, with mating coinciding with the decreasing photoperiod in the autumn, and with calving in the spring. Little is known regarding the factors that influence reproduction in reindeer or of their reproductive physiology. Studies carried out to date have mainly focused on issues related to the population dynamics of wild populations and semi-domestic herds, and to a limited extent on the reproductive physiology of the female. Nor is much known about reproductive disorders and their medical treatment, or of the possibilities to manipulate or control reproduction by the use of hormones. Modern reproductive techniques such as artificial insemination and in vitro fertilisation, maturation and transfer of embryos have so far received scant attention.In the future, it is possible that reindeer under certain conditions might be kept in more intensive production systems. Limited access to high-quality winter pastures and increased demands for productivity have resulted in artificial feeding becoming a common practice in various reindeer herding areas in Scandinavia. In efforts to enhance the productivity of reindeer herds, attention has been focused on factors affecting reproduction in the female and survival of the offspring. Further knowledge on these issues seems necessary when developing strategies for optimalization of meat production in domestic herds and the harvesting of wild populations. This paper puts a broad focus on various aspects of reproduction, including factors influencing the fecundity of reproductively active females. In order to understand these effects it is important also to have a basic understanding of the reproductive physiology of these animals. PMID- 10844225 TI - Reproduction in mustelids. AB - This paper is an attempt to review in a comprehensive manner the reproduction in Mustelidae, based mainly on our own experience of breeding and investigating in captivity various Mustelidae species, mostly of European and Asian origin. Literature data on reproduction in other Mustelidae species used as reference. PMID- 10844226 TI - Reproduction in Old World camels. AB - The article reviews aspects of the basic reproductive biology of the camel, such as puberty, breeding season, ovarian dynamics, synchronisation of ovarian activity and artificial insemination, and superovulation and embryo transfer. Pregnancy and parturition are also discussed. PMID- 10844227 TI - The reproductive pattern and efficiency of female buffaloes. AB - Buffaloes play a prominent role in rural livestock production, particularly in Asia. Reproductive efficiency is the primary factor affecting productivity and is hampered in female buffalo by (i) inherent late maturity, (ii) poor estrus expression in summer, (iii) distinct seasonal reproductive patterns, and (iv) prolonged intercalving intervals. Ovarian function is central to these issues; hence, the focal point of this review is ovarian function in Bubalus bubalis, particularly, in relation to seasonal changes. Ovarian anatomy, follicular and luteal development development, and hormonal profiles during the estrous cycle are discussed. Review of the literature revealed a paucity of critically derived information on follicular and ovulatory patterns in buffalo, particularly, in relation to seasonal estrus/birthing. Efforts may be directed at understanding the process (recruitment, development, atresia) and temporal pattern (follicle selection, dominance, subordinate follicle suppression, follicle numbers, and, preovulatory changes) of follicular dynamics using techniques which permit serial assessment of changes occurring over time. Emphasis may be directed towards investigating follicular "waves" as a functional unit, rather than the estrous cycle, in the context of whole animal endocrinology. The data obtained from such basic studies may then be used to develop and test models for enhancing reproductive efficiency. PMID- 10844228 TI - Epidemiology of reproductive performance in dairy cows. AB - The objectives of this presentation are to review results of our previous and on going research with respect to the risk factors and consequences of poor reproductive performance in dairy cows, and to develop an economic framework to optimize decisions related to dairy cow reproductive performance. To make profitable breeding and replacement decisions, the farmer must account for factors including age, production level, lactation stage, pregnancy status, and disease history of the cows in the herd. Establishing the interrelationships among disease, milk yield, reproduction, and herd management is necessary for developing a decision model for disease treatment, insemination, and replacement. The data for the studies reviewed in this presentation incorporate health, production, and management components from Holsteins in the Northeast USA and Ayrshires from Finland. Data were analyzed using the Cornell Theory Center Supercomputer. The effect of risk factors on reproductive disorders was modeled with logistic regression, and on conception, insemination, and culling with survival analysis. The effect of reproductive disorders on milk yield was analyzed with mixed models. Economic optimization of reproductive performance was done with dynamic programming (DP). High milk yield, high parity, and calving in winter were risk factors for several reproductive disorders. These disorders, in turn, delayed insemination and conception in dairy cows, and some of them increased the risk of culling. Dystocia, retained placenta, and early metritis led to a short-term drop in milk production. High milk yield was not a major factor in delaying conception, except in first parity cows. However, higher yielders were more likely to be inseminated, and less likely to be culled. Non pregnant cows had a higher risk of being culled. Reproductive performance of dairy cows influenced a herd's profitability, and good heat detection and conception rates provided opportunities for management control. It was not always economically advantageous to get cows pregnant as soon as possible, and there was no one optimal value for the calving interval length for all cows in a herd. PMID- 10844229 TI - The effects of bovine viral diarrhoea virus on cattle reproduction in relation to disease control. AB - Bovine viral diarrhoea virus (BVDV) is a major reproductive pathogen in cattle. Infection of the bull can lead to a fall in semen quality and the isolation of infectious virus in the ejaculate, while infection in the cow leads to poor conception rates, abortions and congenital defects. BVDV also reduces the animal's resistance to other respiratory and enteric pathogens. The prevalence of BVDV is primarily due to the efficiency with which the virus crosses the placenta of susceptible females. Calves that survive infection during the first trimester of pregnancy are born with a persistent and lifelong infection. These persistently infected (PI) animals represent between 1.0% and 2.0% of the cattle population and continuously shed infectious virus. The availability of reliable diagnostic ELISA and PCR techniques, which can test milk or serum samples for virus or antibodies, has simplified BVDV surveillance and improved the prospects for control. Although PI animals are the principal vectors within and between herds, they can be readily identified and removed. By contrast, cows carrying a PI foetus are particularly problematic. These animals have been compared to 'Trojan Horses' because they are virus-negative and antibody-positive but they deliver PI calves. In general, acutely infected cattle are much less efficient vectors but infections at the onset of puberty have resulted in a localised and persistent infection within the testes. Under these circumstances, virus shedding into the semen may remain undetected. Transmission of BVDV can be controlled through vaccination or eradication. BVDV vaccine technology has been developing over the past 30 years, but currently available vaccines are still of the conventional inactivated or attenuated sort. In general, vaccination has not been applied with sufficient rigor to make a significant impact on the level of circulating virus, unlike the national and regional eradication programmes established in areas such as Scandinavia, Austria, the Netherlands and Scotland. Eradication confers the added advantage of improved herd health; however, it also creates a susceptible cattle population that needs to be protected by stringent biosecurity. In this article, we discuss how BVDV influences reproductive function, the potential for viral transmission during breeding and the measures that must be taken to avoid the spread of infection to susceptible cattle populations via semen, embryos, culture fluids and infected cows. PMID- 10844230 TI - Epidemiologic concerns relative to in vivo and in vitro production of livestock embryos. AB - Evidence indicates low potential for transmission of pathogens with in vivo derived embryos of cattle when appropriate precautions are taken. In apparent contrast, results of research with in vivo-derived embryos of small ruminants and swine and with in vitro-derived embryos of cattle suggest a greater tendency for their association with pathogens after artificial exposure. However, regardless of donor species, investigations involving collection of embryos from artificially or naturally infected animals and assessment of health of recipients and offspring after transfer of these embryos have indicated low potential for transmitting disease. In this paper, results of embryo-pathogen research are summarized, emphasizing potential for spread of pathogens under natural circumstances. Also, safe embryo handling practices and their application to multiple species are discussed. PMID- 10844231 TI - Bluetongue and equine viral arteritis viruses as models of virus-induced fetal injury and abortion. AB - A number of viruses have the capacity to cross the placenta and infect the fetus to cause, among other potential outcomes, developmental defects (teratogenesis), fetal death and abortion. Bluetongue virus (BTV) infection of fetal ruminants provides an excellent model for the study of virus-induced teratogenesis. This model has shown that only viruses modified by passage in cell culture, such as modified live virus vaccine strains, readily cross the ruminant placenta, and that the timing of fetal infection determines the outcome. Thus, cerebral malformations only occur after fetal infection at critical stages during development and the precise timing of fetal BTV infection determines the severity of the malformation present at birth. Fetal BTV infection also can result in fetal death, followed by abortion or resorption, growth retardation, or no obvious abnormalities, depending on age of the conceptus at infection. Equine arteritis virus (EAV) infection of the equine fetus causes fetal death and abortion but not teratogenesis. These two fetal viral infections are useful not only for the study of teratogenesis and fetal disease, but also to further characterize and compare the complex process that is responsible for normal induction of parturition in ruminants and horses. PMID- 10844233 TI - Relationship of seminal traits and insemination time to fertilization rate and embryo quality. AB - The nature of subfertility due to the male or inseminate is as complex as that of the female. Fertilization failure or failure in embryogenesis are both documented to be of seminal origin. Males also differ in the numbers of sperm required to reach their maximum fertilization rate. Males requiring more sperm would be considered to have compensable seminal deficiencies. These include a number of known (viability and morphology) and unknown factors (functional or molecular traits) precluding sperm access to the ovum or ability to engage the ovum sufficiently to initiate fertilization and the block to polyspermy. Differences in fertility among males or inseminates independent of sperm dosage are considered uncompensable. These deficiencies would be associated with fertilizing sperm that are incompetent to maintain the fertilization process or subsequent embryogenesis once initiated, with most failures occurring prior to maternal recognition of pregnancy. Such sperm would preempt fertilization by competent sperm. Chromatin aberrations in morphologically normal or near normal spermatozoa from abnormal semen samples appear to be the best candidates for the uncompensable deficiency. However, recognition of uncompensable or incompetent fertilizing sperm has not been achieved. Six-day-old non-surgically recovered bovine ova/embryos have been used to evaluate compensable and uncompensable seminal deficiencies as well as to test reproductive strategies. These ova/embryos provide information on fertilization status and embryo quality as well as quantitative and qualitative data regarding associated accessory sperm. Thus, they permit the separation of reproductive failure by fertilization from that by embryonic development. Accessory sperm number is positively associated with both fertilization rate and embryonic quality. Early insemination results in low fertilization rates (low accessory sperm number), but good embryo quality, whereas, late insemination results in high fertilization rates (high accessory sperm number), but poor embryo quality. Additional studies will be necessary to substantiate this model; however, if true, future research designed to improve results to artificial insemination should be tested by breeding early in estrus where sperm viability is most limiting and embryo quality is best. PMID- 10844234 TI - The early fetal life of the equine conceptus. AB - This paper will discuss development of the equine conceptus, especially from the perspective of the maternal environment in which it develops and to which it has considerable influence. PMID- 10844232 TI - Oocyte-sperm interactions. AB - The penetration of the zona pellucida is a crucial step during fertilization. Spermatozoa that are unable to recognize and bind to the zona pellucida glycoproteins or respond to the zona pellucida by undergoing the acrosome reaction fail to fertilize the egg. In most mammalian species, after entering the fallopian tube sperm are stored in the isthmic part of the oviduct under conditions that maintain sperm viability and synchronize both sperm transport and the process of acquisition of fertilizing ability, called capacitation. Only capacitated sperm are enabled to recognize the oocyte and respond to the oocyte signals in an appropriate manner. Close to time of ovulation sperm are released from the oviductal epithelium and swim to site of fertilization. The oviduct and the oocyte itself appear to coordinate sperm function and gamete interaction. The gamete recognition and the next levels of interaction are probably granted by the carbohydrate-protein interactions. Upon binding the signal cascade leading to acrosomal exocytosis is activated, eventually initiated by aggregation of zona pellucida receptor molecules. These signal transducing mechanisms are primed during the capacitation process. Tyrosine phosphorylation, tightly connected to the cholesterol efflux from the plasma membrane, and hyperpolarization seem be involved in this priming by activation of Ca(2+) pathways. Further preparational steps of the acrosome reaction may be mediated by osmosensitive signal transducing mechanisms. The current perspective focuses on the molecules involved in the complex hierarchy of sperm-egg interactions and regulative events priming sperm cell during capacitation for the acrosome reaction. PMID- 10844235 TI - Comparative aspects of equine embryonic development. AB - The developmental changes in the equine conceptus, its maternal environment and their interaction during the first 4 weeks following fertilization are reviewed. Attention is drawn to species-specific events to show why the horse is such a valuable model in which to study early pregnancy. PMID- 10844236 TI - Reproductive management of postpartum cows. AB - High reproductive efficiency in the dairy cow requires a disease-free transition period, high submission rates to AI and high pregnancy rates per service. A key risk factor that causes increased incidence of metabolic disease is low negative energy balance (NEB) in the periparturient and early postpartum periods. Low NEB decreases LH pulse frequency, growth rate and diameter of dominant follicle (DF), IGF-I, glucose, insulin concentrations and increases GH and certain blood metabolites; these effects result in greater loss of body condition score (BCS) and a higher percent of anoestrous cows in the herd. It is important to decrease the incidence of metabolic disease by achieving high dry matter intake (DMI) and minimising the period of NEB after calving. Thus, nutritional management of the cow in the transition period has a crucial role to play in improving reproductive efficiency, because acute nutritional deprivation of heifers has immediate deleterious effects on follicular growth and ovulation. To obtain high submission rates, it is necessary to decrease the incidence of anoestrus and to have good oestrous detection rates. Pregnancy rates per service are affected by a variety of factors. NEB can have deleterious effects on the follicle or the corpus luteum (CL) by decreasing IGF-I concentrations and steroidogenesis. High protein diets fed to postpartum cows leads to increased blood urea and lower fertility. Although the mechanism is not clear, the practical implication of feeding the appropriate level of crude protein in the diet is clear. Thus, a coordinated management approach involving herd managers, nutritionists and veterinarians is required to obtain high reproduction efficiency in dairy cows. PMID- 10844237 TI - Automated electronic systems for the detection of oestrus and timing of AI in cattle. AB - For the majority of dairy herds where artificial insemination (AI) is practiced, the limiting factor toward obtaining efficient reproductive performance is the failure to detect oestrus in a timely and accurate manner. Periodic visual observation has been the dominant method used to identify cows in oestrus. New approaches are being developed to provide automated systems of detection of oestrus using electronic technology. The goal of an oestrus detection program should be to identify oestrus positively and accurately in all cycling animals and consequently to identify animals not cycling. The ultimate goal should be to predict the time of ovulation, thus allowing for insemination that will maximize the opportunity for conception. Unfortunately, most studies designed to evaluate the optimal time of AI generally contained two technical deficiencies: inadequate numbers of cows for valid statistical comparisons and inaccurate knowledge of the onset of oestrus because of low frequency of visual observations and/or efficiency of methods used for the detection of oestrus. Studies using pedometry and a pressure sensing radiotelemetric system will be reviewed as each have independently obtained an optimal time of AI of 5 to 17 h after either the increase in locomotive activity or following the first standing event associated with the onset of oestrus. PMID- 10844238 TI - The use of databases to manage fertility. AB - Dairy farming now needs more records to be kept for quality assurance as well as for management. Herd fertility management is best brought about through the use of computerised records for each animal that integrate fertility, health and production. The development of dairy information systems over the last 25 years has allowed the creation of databases that give rise to "standards" of performance and "interference levels". These databases are of limited use for research unless the coding system has a structure and definition that works across herds. There is an increasing need to incorporate carefully coded disease records into these databases as there is increasing concern about welfare, zoonoses, assurance and the environment. Rules can be determined for satisfactory fertility so interference at an early stage is cost-effective. Integrated indices have been developed (using databases) that incorporate the costs of wastage caused by poor fertility, thus highlighting the priorities for management. Databases are best operated near to the farm, either in the veterinarian's office or on-line in the farm office. Databases can be made into expert systems that deliver high standards of fertility management. A checklist is included that can be followed to analyse the causes of poor fertility in a dairy herd. PMID- 10844239 TI - What is stress, and how does it affect reproduction? AB - Stress is revealed by the inability of an animal to cope with its environment, a phenomenon that is often reflected in a failure to achieve genetic potential. Field data from dairy cows show that stressors such as milk fever or lameness increase the calving to conception interval by 13-14 days, and an extra 0.5 inseminations are required per conception. We suggest that a variety of endocrine regulatory points exist whereby stress limits the efficiency of reproduction. Transport produces an immediate constant increase in arginine vasopressin (AVP) and corticotrophin-releasing hormone (CRH) secretion in ewes, but adrenocorticotrophic hormone (ACTH) reaches a maximum in the first hour while cortisol is highest during the second hour. In contrast, after an insulin injection, the hypothalamo-pituitary-adrenal (HPA) response is delayed occurring only after glucose decreases below a threshold. Changes in AVP, CRH and ACTH each follow a similar time course, but eventually the secretion of AVP and CRH decreases while glucose is still at a nadir. Negative feedback effects appear to operate mainly at the pituitary level during transport but at the hypothalamus during hypoglycaemia. We also have endocrine evidence to show that stressors interfere with precise timings of reproductive hormone release within the follicular phase. Transport, or insulin, reduce the frequency and amplitude of gonadotrophin-releasing hormone and LH pulses, suggesting that these stressors exert effects at the hypothalamus or higher centres in the brain. Both stressors also delay the onset of the luteinising hormone (LH) surge. Preliminary results suggest that opioids mediate these effects but progesterone/glucocorticoid receptors are not involved because the antagonist, RU486, is unable to reverse insulin-induced delays in the LH surge. There is also evidence to support effects at pituitary level because exogenous ACTH, or transport, reduce the amount of LH released by challenges with GnRH. The reduction in endogenous GnRH/LH secretion ultimately deprives the ovarian follicle of adequate gonadotrophin support leading to reduced oestradiol production by slower growing follicles. Thus, there is a level of interference by stressors at the ovary. Reproduction is such an important physiological system that animals have to ensure that they can respond to their surroundings; thus, it is advantageous to have several protein mechanisms, i.e. at higher brain, hypothalamus, pituitary and target gland levels. However, when pushed too far, subfertility occurs. PMID- 10844240 TI - Database of mouse strains carrying targeted mutations in genes affecting cellular responses to DNA damage. Version 4. PMID- 10844241 TI - Enhanced Tn10 and mini-Tn10 precise excision in DNA replication mutants of Escherichia coli K12. AB - The precise excision of transposon Tn10 and a mini-Tn10 derivative, inserted in the gal or lac operons, was studied in dnaB252 and dnaE486 temperature-sensitive mutants of Escherichia coli. dnaB codes for a DNA replication helicase and dnaE for the alpha subunit of DNA polymerase III. Mutations in these genes were found to enhance, at the permissive temperature, the precise excision of both genetic elements. The increase factor was much more pronounced for the dnaB252 mutant with the transposons inserted in gal. The stimulated excision was only partially affected by a recA null mutation but was significantly reduced by introduction of recF null or ruvA mutations. A model involving template switching of the polymerase between the direct repeats flanking the transposons, on the same strand or between sister strands, could account for the observed results. PMID- 10844242 TI - Decreased UV sensitivity, mismatch repair activity and abnormal cell cycle checkpoints in skin cancer cell lines derived from UVB-irradiated XPA-deficient mice. AB - Xeroderma pigmentosum group A gene (XPA)-deficient mice are defective in nucleotide excision repair (NER) and are therefore highly sensitive to ultraviolet (UV)-induced skin carcinogenesis. We established cell lines from skin cancers of UVB-irradiated XPA-deficient mice to investigate the phenotypic changes occurring during skin carcinogenesis. As anticipated, the skin cancer cell lines were devoid of NER activity but were less sensitive to killing by UV irradiation than the XPA(-/-) fibroblast cell line. The lines were also more resistant to 6-thioguanine (6-TG) than XPA(-/-) and XPA(+/+) fibroblasts, which was suggestive of a mismatch repair (MMR) defect. Indeed, in vitro mismatch binding and MMR activity were impaired in several of these cell lines. Moreover, these cell lines displayed cell cycle checkpoint derangements following UV irradiation and 6-TG exposure. The above findings suggest that MMR downregulation may help cells escape killing by UVB, as was seen previously for methylating agents and cisplatin, and thus that MMR deficient clones are selected for during the tumorigenic transformation of XPA(-/-) cells. PMID- 10844243 TI - Treatment of mammalian cells with mimosine generates DNA breaks. AB - Exponentially growing mouse erythroleukemia (MEL) cells and quiescent human peripheral blood lymphocytes (PBL) were treated with different concentrations of the nonprotein amino acid mimosine for 16 h. The treatment of the cycling cell population with 400 microM mimosine caused inhibition of DNA replication, changes in the progression of the cells in the cell cycle, and apoptosis. Nucleoid sedimentation analysis and comet assay were used to monitor the appearance and accumulation of DNA breaks. The rate of break accumulation was dose-dependent, did not depend on the stage of the cell cycle and was not connected with the mechanism of DNA replication. The data indicate that the effects of mimosine on DNA synthesis and the cell cycle may be a result of introduction of breaks into DNA. PMID- 10844244 TI - Purification and characterization of a novel UV lesion-specific DNA glycosylase/AP lyase from Bacillus sphaericus. AB - The purification and characterization of a pyrimidine dimer-specific glycosylase/AP lyase from Bacillus sphaericus (Bsp-pdg) are reported. Bsp-pdg is highly specific for DNA containing the cis-syn cyclobutane pyrimidine dimer, displaying no detectable activity on oligonucleotides with trans-syn I, trans-syn II, (6-4), or Dewar photoproducts. Like other glycosylase/AP lyases that sequentially cleave the N--glycosyl bond of the 5' pyrimidine of a cyclobutane pyrimidine dimer, and the phosphodiester backbone, this enzyme appears to utilize a primary amine as the attacking nucleophile. The formation of a covalent enzyme DNA imino intermediate is evidenced by the ability to trap this protein-DNA complex by reduction with sodium borohydride. Also consistent with its AP lyase activity, Bsp-pdg was shown to incise an AP site-containing oligonucleotide, yielding beta- and delta-elimination products. N-terminal amino acid sequence analysis of this 26 kDa protein revealed little amino acid homology to any previously reported protein. This is the first report of a glycosylase/AP lyase enzyme from Bacillus sphaericus that is specific for cis-syn pyrimidine dimers. PMID- 10844245 TI - Isolation of juvenile hormone esterase and its partial cDNA clone from the beetle, Tenebrio molitor. AB - Juvenile hormone esterase (JHE) plays an essential role in insect development. It is partially responsible for the clearance of juvenile hormone (JH) which regulates various aspects of insect development and reproduction. Because of its role in regulating JH titer, this enzyme has been targeted for development of biologically-based insecticides. JHE was partially purified from the beetle, Tenebrio molitor, using a transition state analog as the affinity ligand. Two forms of JHE were characterized by activity analysis, isoelectric focusing, two dimensional SDS-PAGE and N-terminal sequence analysis. The esterase is associated with two proteins of sizes 71 and 150 kDa, both of which are active on JH III. A partial cDNA clone for the enzyme was isolated based on the sequence of N terminal and internal peptides. Its sequence indicates that JHE from T. molitor and Heliothis virescens may have a common origin. PMID- 10844246 TI - A dual role of 20-hydroxyecdysone in the control of protein synthesis related to DNA puff activity in the anterior region of Bradysia hygida (Diptera, Sciaridae) salivary gland. AB - During the last 30 h of the larval stage, the salivary glands of Bradysia hygida show the amplification of some genes, resulting in the formation of two successive groups of DNA puffs, which direct the synthesis of two different sets of polypeptides. Incubation of anterior (S1) salivary gland regions, at age E7, beginning of first group of DNA puffs activity, in culture medium for 2 to 10 h results in a decrease in the synthesis of the polypeptides characteristic of this period. However, during subsequent incubation (from E7 to E7+12 h-24 h), when the second group of DNA puffs is active, S1 regions were able to synthesize some polypeptides characteristic of this period. The role of 20-OH ecdysone was studied, in vitro and in vivo, during these two periods of protein synthesis in S1 regions. The presence of the hormone was shown to be necessary to maintain, in vitro, the synthesis of the first set of polypeptides and was strongly inhibitory, in vitro and in vivo, to the synthesis of the second set of polypeptides. Thus, it is likely that the activity of the two distinct groups of DNA puffs is under opposite 20-OH-ecdysone control mechanisms. PMID- 10844247 TI - Synthesis of vitellogenin by the follicle cells of Rhodnius prolixus. AB - The synthesis and secretion of vitellogenin by the ovary of Rhodnius prolixus was investigated. Using whole ovary or epithelial cells isolated from follicles of different sizes, it is shown that the follicle cells are a site of synthesis for this protein in the ovary. The ovaries or follicle cells were incubated in vitro with [(35)S]-methionine or (32)Pi and the secretion of newly synthesized ovarian vitellogenin (O-Vg) was estimated by the radioactivity associated with the immunoprecipitate or acid-precipitate proteins in the culture medium. The radioactive O-Vg was analyzed by SDS-PAGE followed by autoradiography or after elution from a DEAE-Toyopearl column. The presence of O-Vg inside the follicle cells was detected by immunofluorescence and immunogold labels. Both methods revealed strong labeling inside the follicle cells. While the capacity for total protein synthesis by the follicle cells was maximal during the early phase of vitellogenesis (in small follicles), the synthesis of O-Vg reached its peak during the late phase of oocyte growth, just before formation of the chorion. A possible role for ovarian vitellogenin in Rhodnius and its relationship with Vg synthesis by the fat body is discussed. PMID- 10844248 TI - Inducible P450s of the CYP9 family from larval Manduca sexta midgut. AB - Several related cytochrome P450 cDNAs belonging to the CYP9 family have been cloned from the midgut of larval tobacco hornworms, Manduca sexta. The first P450, CYP9A2, was obtained by RT-PCR using degenerate primers. Northern blot analysis of expression in the midgut using the CYP9A2 probe revealed a significant induction by a variety of chemicals. Diets supplemented with the wild tomato compound 2-undecanone caused a dose-dependent induction which peaked after 48 h. Induction was also observed after addition to the diet of indole-3 carbinol, phenobarbital, 2-tridecanone and xanthotoxin. Neither alpha-pinene, clofibrate nor nicotine were effective inducers. The CYP9A2 probe hybridized to two mRNA species, one of 2. 0 kb and another of 4.2 kb, suggesting cross hybridization to other P450 mRNAs. Additional P450 clones of the CYP9 family were then obtained and sequenced. Northern hybridization revealed that the 4.2 kb band also hybridized to CYP9A4 whereas the 2.0 kb hybridized to CYP9A5. Despite being 91% identical, CYP9A4 and CYP9A5 were induced differentially by clofibrate and xanthotoxin. Multiple P450 genes from various families are therefore induced in Lepidoptera in response to plant allelochemicals or xenobiotics. PMID- 10844249 TI - Studies on proteins in post-ecdysial nymphal cuticle of locust, Locusta migratoria, and cockroach, Blaberus craniifer. AB - Proteins were extracted from the cuticle of mid-instar nymphs of locusts, Locusta migratoria, and cockroaches, Blaberus craniifer. Seven proteins were purified from the locust extract and five from the cockroach extract, and their amino acid sequences were determined. Polyacrylamide gel electrophoresis indicates that the proteins are present only in the post-ecdysially deposited layer of the nymphal cuticles. One of the locust and one of the cockroach nymphal proteins contain a 68-residue motif, the RR-2 sequence, which has been reported for several proteins from the solid cuticles of other insect species. Two of the cockroach proteins contain a 75-residue motif, which is also present in a protein from the larval/pupal cuticle of a beetle, Tenebrio molitor, and in proteins from the exoskeletons of a lobster, Homarus americanus, and a spider, Araneus diadematus. The motif contains a variant of the Rebers-Riddiford consensus sequence, and is called the RR-3 motif. One of the locust and three of the cockroach post-ecdysial proteins contain one or more copies of an 18-residue motif, previously reported in a protein from Bombyx mori pupal cuticle. The nymphal post-ecdysial proteins from both species have features in common with pre-ecdysial proteins (pharate proteins) in cuticles destined to be sclerotised; they show little similarity to the post-ecdysial cuticular proteins from adult locusts or to proteins from soft, pliable cuticles. Possible roles for post-ecdysial cuticular proteins are discussed in relation to the reported structures. PMID- 10844251 TI - Characterization of subclones of the epithelial cell line from Chironomus tentans resistant to the insecticide RH 5992, a non-steroidal moulting hormone agonist. AB - Selection of hormone resistant subclones in the continuous presence of the insecticide and ecdysteroid mimick RH 5992 (tefubenozide) resulted preferentially in clones with defects in ecdysteroid receptor function. RH 5992 is already degraded to polar products in wild-type cells; no increase in metabolism of tefubenozide is observed in resistant clones. According to Western blots, ecdysteroid receptor (EcR) and its heterodimerization partner ultraspiracle (USP) are present in all resistant clones. The concentrations are comparable to wild type cells, but in three clones the extent of phosphorylation of USP is diminished. With regard to hormone binding several types of hormone resistance are distinguished: (1) The same two high-affinity hormone recognition sites are present as in wild-type cells (K(D1)=0.31+/-0.28 nM, K(D2)=6.5+/-2.4 nM) but the number of binding sites is reduced. (2) The binding site with the lower affinity (K(D2)) is missing. (3) The binding site with the higher affinity (K(D1)) is missing. (4) No specific binding is observed. Ponasterone A binding can be rescued by addition of EcR but not by USP. (5) Ligand specificity is altered. RH 5992 can not compete [(3)H]-ponasterone A as efficient as in wild-type cells. PMID- 10844250 TI - cDNA cloning and expression of a hormone-regulated heat shock protein (hsc 70) from the prothoracic gland of Manduca sexta. AB - The brain neuropeptide prothoracicotropic hormone (PTTH) stimulates a rapid increase in ecdysteroid hormone synthesis that is accompanied by general and specific increases in protein synthesis, including that of a 70 kDa cognate heat shock protein (hsc 70). To further understand the possible roles of hsc 70, hsc 70 cDNA clones were isolated from a tobacco hornworm (Manduca sexta) prothoracic gland cDNA library. All sequenced clones were highly homologous to the Drosophila hsc 70-4 isoform. Manduca hsc 70 mRNA levels during the last larval instar exhibited a peak at the onset of wandering and a peak that coincided with the major pre-metamorphic peak of ecdysteroid synthesis. Manipulations of the glands' hormonal milieu showed that hsc 70 mRNA levels respond to 20-hydroxyecdysone, dibutyryl cAMP, PTTH and the JH analogue hydroprene. The protein and mRNA data suggest that hsc 70 could be involved in a negative feedback loop regulating assembly of the ecdysone receptor complex. PMID- 10844252 TI - Production, regeneration and biochemical precursors of the major components of the defensive secretion of Eurycotis floridana (Dictyoptera, polyzosteriinae). AB - The defensive secretion of the cockroach Eurycotis floridana contains three main components, (E)-2-hexenal, (E)-2-hexenol and (E)-2-hexenoic acid, which represented about 98% of the organic phase. The quantity of the aldehyde, alcohol, and acid present in the defensive secretion increased rapidly for 60 days from the imaginal moult. Following artificial discharge, the males were able to regenerate their initial volume of secretion over a 30 day period. To investigate the possible routes of biosynthesis of the three components, E. floridana was injected with 14C-labeled fatty acids and acetate, and the incorporation of 14C into the three components were quantified 1, 6, and 24 h after milking. Our results revealed that oleic, linoleic, linolenic and palmitic acids, which constitute part of the fat body of the insect, were incorporated to the same degree into the three main components, but very slowly compared to acetate. Although it has not been possible to identify the exact route of aldehyde, alcohol and acid biosynthesis, our findings suggests that (E)-2 hexenal, (E)-2-hexenol and (E)-2-hexenoic acid are preferentially biosynthesized de novo from acetate. PMID- 10844254 TI - [Reasonable hospital use of mupirocin]. PMID- 10844255 TI - [Vulvodynia]. PMID- 10844256 TI - [Evaluation of the understanding of sun risk among 241 adolescents]. AB - OBJECTIVE: We investigated adolescent's knowledge of the consequences of sun exposure in order to determine what preventive measures would be effective. MATERIAL AND METHODS: A questionnaire was given to 241 adolescents (111 girls and 130 boys) aged 13 to 15 years whose background included all social occupational categories. The questionnaire included 21 open response items. We also interviewed 100 adolescents 3 months after an information session with a 1-hour slide presentation. RESULTS: Eighty-five percent of the girls and 62 p. 100 of the boys said they exposed themselves to sun regularly. For 50 p. 100 of the adolescents, sun exposure occurred between 12 noon and 4 p.m. Less than 9 p. 100 knew that this period corresponds to maximal sunlight intensity and that maximal photoprotection is necessary. Only 47 p. 100 of the girls and 25 p. 100 of the boys regularly used protective skin lotions. Twenty-five percent of the girls used a lotion adapted to their phototype and 8 p. 100 of the boys. Twenty percent of the adolescents questioned were aware that applications should be repeated every 2 hours or after bathing. The cost of skin lotions was never cited as a limiting factor for their use. More than 90 p. 100 of the adolescents gave correct responses at the second test. DISCUSSION: Our study demonstrated that the risk of sun exposure is largely unrecognized by adolescents and that they have very incomplete information concerning protective measures. An adapted information program would be useful for this risk population. PMID- 10844257 TI - [Vulvodynia and facticious urticaria]. AB - BACKGROUND: The term vulvodynia describes a multifactorial syndrome of chronic vulvar discomfort with burning, itching, pain, and dyspareunia. Due to the paucity of clinical signs, this syndrome is often perceived as a predominantly psychiatric problem. A context of symptomatic dermographism has been reported in rare instances. PATIENTS AND METHODS: In our interdisciplinary consultation of vulvar dermatology, we routinely look for the presence of facticious urticaria. In the last 17 months, we observed 14 patients with an association of facticious urticaria and vulvodynia. RESULTS: The principal symptoms were itching and burning. Sexual intercourse, tight clothes, and stress were most often cited as aggravating factors. The efficacy of a systemic antihistaminic treatment observed in our patients suggests that certain cases of vulvodynia might be an expression of symptomatic dermographism. DISCUSSION: Therefore, a check-up for chronic vulvar discomfort should comprise a test for the presence of facticious whealing. The prevalence of a causal relationship between chronic vulvar discomfort and facticious whealing remains to be established. PMID- 10844258 TI - [Chronic desquamative gingivitis syndrome: retrospective analysis of 33 cases]. AB - OBJECTIVE: Desquamative gingivitis is a chronic diffuse inflammation of the gingiva. The aim of this study was to determine the causes and the clinical characteristics of desquamative gingivitis. PATIENTS AND METHODS: This was a retrospective descriptive study including 33 consecutive patients (25 women and 8 men) seen at a dermatology clinic for erosive gingivitis. RESULTS: Thirteen patients (39 p. 100) had cicatricial pemphigoid, 12 (36 p. 100) had lichen planus, and 5 (15 p. 100) had pemphigus. Delay to diagnosis was a mean 19 months. The pinch sign was positive in 12 of the 13 cases of cicatricial pemphigoid. Dapsone improved the buccal lesions of cicatricial pemphigoid in all cases. Systemic corticosteroid therapy and acitretine were the most effective treatments for lichen planus and corticosteroid therapy improved pemphigus in all cases. At the time of assessment, only 3 cases of cicatricial pemphigoid, 2 cases of lichen planus and 1 case of pemphigus had reached complete remission without treatment. DISCUSSION: Cicatricial pemphigoid and lichen planus are the most frequent causes of desquamative gingivitis, accounting for three-quarters of the cases. Positive diagnosis may be difficult and may require sophisticated techniques to avoid delay. Despite the effectiveness of symptomatic treatment, desquamative gingivitis may have a long course. PMID- 10844260 TI - [Cutaneous actinomycosis in the perianal area and buttocks]. AB - INTRODUCTION: Actinomycosis is a rare spreading suppurative and granulomatous disease. In 50 to 60 p. 100 of cases, cervico-facial area is involved. Involvement of other cutaneous areas is very uncommon. CASE-REPORTS: Two men (51 and 58 year-old, respectively) were admitted to the hospital because of erythematous and violaceous nodules with fistulization to the skin, localized on the perianal area and buttocks for several months. Bacterial cultures of the pus and of skin biopsy specimens yielded Actinomyces meyeri in case 1 and Actinomyces israelii in case 2. Patients did not have other infectious lesions. Treatment included systemic antibiotics and surgical iterative excisions in case 1. DISCUSSION: The main interest of those two cases is their similar clinical presentation and in one case the rarity of the micro-organism involved (Actinomyces meyeri). Since the lesions involved only the skin, we postulate that the organism may have extended from the oral mucosa through the gastrointestinal tract. PMID- 10844259 TI - [Dermatomyositis and nasopharyngeal carcinoma: 3 cases]. AB - BACKGROUND: Nasopharyngeal carcinoma is a common cancer in Tunisia with an estimated incidence of 1.8/100,000. The tumor shows a characteristic association with paraneoplastic syndromes. CASE REPORTS: We report three cases of histologically proven dermatomyositis associated with nasopharyngeal carcinoma in patients aged 40, 24 and 65 years. In all cases, the course of the paraneoplastic syndrome ran parallel to the nasopharyngeal carcinoma. DISCUSSION: Dermatomyositis is one of the numerous paraneoplastic syndromes associated with nasopharyngeal carcinoma. Systematic examination of the nasopharynx is required in case of dermatomyositis. PMID- 10844261 TI - [Human orf and erythema multiforme]. AB - BACKGROUND: Orf is a disease of sheep due to a parapoxvirus. Transmission in man is possible, and is generally benign, self-limited condition. Although, rare complications have been mentioned. We report a case of Orf with erythema multiforme in a young girl. CASE REPORT: A 13-year-old girl presented to our department with an erythema multiform. The cutaneous examination revealed painless erythematous nodule of her right index finger developed 15 days after a contact with a lamb. The diagnosis of Orf complicated by erythema multiforme was made on the basis of her typical history. Lesions heal with symptomatic therapy in 15 days. DISCUSSION: Orf presents in sheeps as a pustular dermatitis or a vesicular oral mucosal lesions, and in man as a single lesion on the site of infection. This lesion generally heals without complications, although secondary infection is not uncommon. Twenty one cases of erythema multiforme have been recorded following infection with the Orf virus. In typical cases, the diagnosis is established by a history of contact with infected animals and the appearance of the lesion. Electron microscopic examination can be helpful sometimes. Lesions regress in 4 to 5 weeks without specific treatment. PMID- 10844262 TI - [Hereditary progressive mucinous histiocytosis]. AB - BACKGROUND: Hereditary progressive mucinous histiocytosis is an uncommon condition which to date has only been observed in women. The disease is a non Langerhans histiocytosis with a stereotypic clinical presentation limited to the skin. The clinical, histological and genetic features of this disease differ considerably from other types of histiocytoses and overload diseases. We report the first case observed in France. CASE REPORT: A 49-year-old woman consulted for asymptomatic papules on the dorsal aspect of the hands known since childhood. The lesions had progressively spread to other regions of the body. The woman's mother and two sisters had the same condition but her two sons and her brother and nephews did not. Standard semi-thin slices and electron microscope histology demonstrated dense cellular infiltration of the reticular dermis with interstitial mucinous overload, voluminous histiocytes with characteristic toluidine blue positive granulations, and a cytoplasm rich in myelin and Zebra bodies respectively. The observed clinical, histological and genetic features were characteristic of progressive hereditary progressive mucinous histiocytosis. DISCUSSION: The pedigree in our case and in those reported in the literature suggest a dominant hereditary condition. The fact that only female cases have been reported to date would suggest X-linked transmission with early death of male fetuses. An autosomal dominant transmission limited to woman as a result of mitochondrial heredity or hormone-related phenomena cannot be ruled out. The progressive extension with age without a tendency for spontaneous regression and the electron microscopy aspect suggest a phospholipid deposit disorder similar to that observed in lysosomial diseases. However, primary macrophage proliferation in response to an unknown stimulus cannot be excluded. PMID- 10844263 TI - [Major eosinophilia in a premature infant induced by topical ketoconazole]. AB - INTRODUCTION: Ketoconazole is a member of a newer group of imidazole antifungals. Except for treatment of oral candidiasis, there is no reported experience of use in neonates. Thus, its use in neonatalogy must be considered experimental. Herein we report a major eosinophilia in a very low birth weight neonate induced by transcutaneous resorption of topical ketoconazole. CASE REPORT: A 26 week premature was intubated and ventilated for a hyaline membrane disease. At more than 2 weeks, he developed diaper dermatitis with Candida albicans. Treatment consisted of local application of ketoconazole. After 6 days of application, WBC showed a major eosinophilia [20,000/microL]. Discontinuation of the drug was followed by a prompt normalization of the eosinophil count. The challenge test by topical ketoconazole reproduced eosinophilia, implicating this drug, which, to our knowledge, has not been described previously to cause eosinophilia in very low weight neonates. DISCUSSION: Peripheral eosinophilia is an uncommon finding present in a relatively limited number of conditions in children. A minor eosinophilia often seen in premature neonates may in part be due to intubation. This case illustrates the potential danger of percutaneous application of drugs in newborn and infants as it has previously been pointed out way various chemicals. Any deterioration of the corneal layer such as observed in many dermatosis prompts an increase in the cutaneous permeability. Immaturity of the premature skin may have aiso played a role. Dermatologists should be aware of potential toxic topical ointments before prescribing them to very low weight babies. PMID- 10844264 TI - [Successful treatment of corticoid-resistant pemphigus with high-dose intravenous immunoglobulins]. AB - INTRODUCTION: Pemphigus vulgaris is a serious autoimmune bullous disease, that may be difficult to control. Although corticosteroids have dramatically improved the outcome of the disease, this treatment may be complicated by unresponsiveness or serious side-effects. We report the case of a patient with pemphigus vulgaris refractory to corticosteroids who responded rapidly to the addition of high-dose intravenous immunoglobulins. CASE REPORT: A 38-year-old man presented with a 1 month history of widespread bullous lesions of the skin and oral mucosa. The diagnosis of pemphigus vulgaris was made on the results of histology and direct immunofluorescence of perilesional skin. Systemic corticosteroids were initially started, but cutaneous and mucosal lesions poorly responded after 6 weeks. Mensual cycled of intravenous immunoglobulins were then begun and led to a complete disappearance of the lesions after three cycles. Four courses of high dose intravenous were administered, that allowed to reduce doses of steroids. The patient was in complete remission without treatment after a two-year follow-up. DISCUSSION: Pulse therapy with high-dose intravenous immunoglobulins has been occasionally used for the treatment of pemphigus vulgaris, especially in an attempt to reduce side-effects of immunosuppressive agents or when these therapies are ineffective. We report an additional case, suggesting in addition of recent data of literature, that immunoglobulins may be useful as an alternative treatment in pemphigus vulgaris. PMID- 10844265 TI - [Cat vitiligo]. PMID- 10844266 TI - [Taxonomy of Leishmania and geographical distribution of leishmaniasis]. PMID- 10844267 TI - [Topical antibiotics used in dermatology: efficiency, indications and adverse events]. PMID- 10844268 TI - [Apoptosis in dermatology]. PMID- 10844269 TI - [Should dermatologists use EMBASE for bibliography searches?]. PMID- 10844270 TI - [Cutaneous expression of peripheral benzodiazepine receptors]. PMID- 10844271 TI - [Alfuzosine-induced dermatomyositis] [In Process Citation] PMID- 10844272 TI - [French-speaking dermatologists and syphiligraphists'meeting - paris, june 6-8, 1922] [In Process Citation] PMID- 10844273 TI - [Strategy against nosocomial infections in France]. AB - Because of their extent and due to the complexity of their management, nosocomial infections (NI) have become a public health problem in view of ethical and economical considerations. A decree, issued on 05.06.1988, organizes the supervision and the prevention of NI throughout the country and requires the setting up of a committee against nosocomial infections (CLIN) in every medical establishment. In 1992 five inter-areas coordination committees (C-CLIN) and one national technical struggle committee (CTIN) were established in France. A governmental plan was instituted in 1994. This plan, enacted in the 04.19.1995 text, focuses on the importance of reducing by 30% the frequency of NI by the year 2000. The creation of a NI cell in the Health Ministry and of operational hygiene teams in the hospitals have since then strenghtened the anti-infection system. The CLIN became a compulsory institution in the 1998 for private establishments and a new decree is to be issued at the end of 1999 and to come into force on 12.31.2001 at the latest. Finally, the decree n degrees 98-535 (07.01.1998) sets up the French Agencies for Health Safety and an Institute of Sanitary Supervision. Both structures are in charge of the surveillance system in Health establishments. The CTIN and the C-CLIN, together with groups of experts and learned societies (Superior Council for Public Hygiene in France, French Society for Hospital Hygiene, Health National Agency of Accreditation and Valuation), will edit official guide of technical recommendations. This ambitions strategy, based on education, information, supervision and prevention methods, requires a multidisciplinary mobilization and needs the compulsory support of the extire hospital community. PMID- 10844274 TI - [Methicillin-resistant Staphylococcus aureus nosocomial infections: epidemiology, prevention and antibiotic therapy]. AB - Infections caused by methicillin-resistant Staphylococcus aureus strains are particularly representative of the various questions raised by nosocomial infection. The specific ecology of these strains, which is limited to care centers, gives evidence to their nosocomial origin. The mode of spread is mainly clonal by cross-transmission, which emphasizes the importance both of individual hygiene measures and procedures for spread control. Both the incidence and percentage of methicillin-resistant strains are tracers of choice for the local status as regards the risk of nosocomial infection. Multiresistance of most methicillin-resistant Staphylococcus aureus strains causes therapeutical difficulties, emphasizing the importance of controlling spread of glycopeptide resistant strains. PMID- 10844275 TI - [Measures for controlling the transmission of hospital bacteria]. AB - Three elemental steps need to be taken in order to control and prevent multiresistant bacteria spreading: screening for BMR carriage - signalling - isolation precautions PMID- 10844276 TI - [Pharmaceutical contribution to the non specific challenge in bronchial provocation tests]. AB - Aerosols of methacholine chloride are used in inhalation tests to measure non specific bronchial responsiveness. Inhalation challenge solutions are not commercially available neither by pharmaceutical industry nor in France by the PCH-APHP. To reply to doctor's request, dilute concentrations of methacholine must be prepared properly in order to protect the patient and to assure doctor of test reproductibility. Osmolarity, pH and degradation of different dilute concentration of methacholine chloride have been compared with two diluents. This study allowed us to validate our preparation of solutions. PMID- 10844277 TI - [Effect of "micronized" C60 fullerene on the microbial growth in vitro]. AB - The effect of C60 "micronized fullerene was tested by a manual and automatised (Analyzer Bioscreen C(R)) micromethod on the microbial growth of 22 collection strains: E. coli (5), P. aeruginosa (2), S. Typhimurium (6), S. aureus (2), L. monocytogenes (2), E. hirae (1), B. cereus (1), B. subtilis (1), B. pumilus (1) et C. albicans (1). No effect on microbial growth was observed with C60 "micronized fullerene (43.2 microg/ml) on all strains studied: no difference was found with doubling time, slope and growth rate constant. The results of cytotoxicity obtained with animal models or in vitro cultures as human monocyte, leukocyte or macrophage confirm the absence of effect of C60 fullerene at a concentration compatible with microbial or biological models. This study is included in research program headed "Therapeutics perspectives of fullerenes. PMID- 10844278 TI - [Validation of a method to inactivate fentanyl in the used devices of Durogesic]. AB - A recent literature study has shown that after 3 days of use of a transdermic patch of fentanyl (Durogesic(R)), there was such an amount of drug remaining that may make its excessive use possible. First of all, we have developed a fentanyl assay by a spectrophotometric method and then, the remaining amount of fentanyl was determined in 29 used patches. This study has shown that a mean of 22% of fentanyl remained in the used patches, each original content astounded. Then, we have determined an easy and feasible method in a chemist's shop allowing inactivation of fentanyl in the used patches and so preventing its diversion. PMID- 10844279 TI - [Synthesis of carbamoyl tetrahydro 1,4,2-dioxazines and evaluation of their activity against human immunodeficiency virus]. AB - 1,4,2-dioxazine derivatives were synthesized from B-ureidoxyalcools, and their 1H, 13C RMN and mass spectra were determined. Their activity against human immunodeficiency virus and their cytotoxicity were then evaluated. Whereas the derivatives had no effect on virus infectivity nor on reverse transcriptase activity, they unexpectedly enhanced host cell infection by both lymphotropic and macrophage-tropic virus strains. PMID- 10844280 TI - [The gene of human prion: reassessment of its features and pathogenic role]. AB - After a detailed examination of some features of the human prion gene, we propose a reassessment of its role in the pathogenicity of the derived neurodegenerative diseases e.g. Creutzfeldt-Jakob's disease. The particular resistance of this gene towards physico-chemical agents is related to the relative abundance of the guanine and cytosine bases. Other features such as the paucity of CpG sequences, GC box, polyadenylic tail, direct and inverted repeats, all evoke those of retroviral LTRs. Finally, the presence of polypurinic and polyadenylic sequences, as well as that of a functional AP2 element coding a superantigen are all reminiscent of a parenthood of this gene with that of the murine virus MMTV, whose pathogenicity is based on the interaction between either endogenous or exogenous mutated viral forms. A similar biological behavior between, respectively the MMTV virus and the endogenous plus exogenous infectious forms of the prion is therefore suggested. All these clues are in agreement with the prominent data of the natural history of prion diseases, but il would be necessary that the prion protein should be associated with a nucleic acid belonging to its own gene and which would play a role in the animal or human illness, together with the endogenous prion gene. Thus, some exogenous forms originating from animals, e.g. bovine or perhaps ovine ones, could recombine with the human prion gene and would be responsible for the human illnesses. PMID- 10844281 TI - [Feasibility of 400 microm spheroids by extrusion-spheronization]. AB - Extrusion-spheronisation technology is an agglomeration process that makes it possible to obtain spheroids. The aim of this work was to obtain 400microm spheroids that could be sprinkled on food to improve patient observance in particular with children and old people. A methodology to select wet masses for extrusion-spheronisation through a 400microm orifice was developed. First, it was nesessary to define the parameters that make it possible to appreciate the qualities that the wet mass and the extrudates have to possess and their method of evaluation: plasticity, cohesiveness, brittleness of the mass and the extrudates, appearance of extrudates. A feasibility assay made on the drum extruder was then performed. After proving the feasibility of 400microm spheroids of Gelucire 50/02 we considered the association of a drug with it. PMID- 10844282 TI - [ [In Process Citation] PMID- 10844283 TI - [Development of laparoscopic biliary surgery in the last decade]. PMID- 10844284 TI - [Laparoscopic cholecystectomy in the treatment of acute cholecystitis. Prospective non-randomized study]. AB - OBJECTIVES: To evaluate the results of laparoscopic cholecystectomy in patients with acute cholecystitis and to determine the optimal timing. PATIENTS AND METHODS: From January 1993 to December 1999, 168 patients (91 women and 77 men with a mean age of 57.3 years) underwent laparoscopic cholecystectomy for acute cholecystitis confirmed by histopathological examination. Patients were divided into 2 groups (similar in age and ASA classification): group 1 (106 patients) underwent laparoscopic cholecystectomy within 3 days after the onset of symptoms of acute cholecystitis and group 2 (62 patients) underwent laparoscopic cholecystectomy after 3 days. RESULTS: There was no death. Conversion rates were 30.9%. Surgery lasted 141.3 min, postoperative morbidity was 12.5%, and the postoperative length of hospital stay was 6.9 days. The conversion rates in patients who underwent surgery before and 3 days after the onset of symptoms were respectively 21.7% and 46.7% (P=0.0007). There was no statistical difference between early and delayed surgery for time to surgery and postoperative complications. On the other hand, the total hospital stay was significantly shorter in group 1. CONCLUSION: Laparoscopic cholecystectomy for acute cholecystitis is safe and associated with a shorter postoperative stay, a lesser morbidity and a lesser mortality compared to "open" surgery. Laparoscopic cholecystectomy should be carried out soon as the diagnostic of acute cholecystitis is established and preferably before 3 days after the onset of the symptoms. Early laparoscopic cholecystectomy allows a reduction of the conversion rate and a reduction of total hospital stay that are medical and economic benefits. PMID- 10844286 TI - [Digestive and hepatic signs of schistosomiasis]. PMID- 10844285 TI - [Treatment of common bile duct stones with laparoscopy. Retrospective multicenter study with 612 patients]. AB - OBJECTIVE: To analyse retrospectively the results of one-stage laparoscopic treatment for common bile duct stones in 19 surgical centers in France. PATIENTS: From January 1991 to July 1996, 612 patients with choledocholithiasis underwent laparoscopic treatment. RESULTS: Overall duct clearance was obtained in 489 of the 612 patients (80%): through the cystic duct in 222 of 380 patients (58.4%), by secondary choledochotomy (after unsuccessful transcystic duct extraction) in 77 of 96 (80%), and in 190 of 232 (82 %) by primary choledochotomy. The overall duct clearance rate increased from 65% in 1991 to 84% in 1996. The use of the choledochotomy approach increased from 43% in 1991 to 69% in 1996 (P<0.01), due to a substantial increase in primary choledochotomy. In contrast, the use of the transcystic approach decreased from 57% to 31% (P<0.01). The mean time for surgery was shorter for cystic duct exploration than for primary choledochotomy (101+/-51 vs. 155+/-62 min, P<0.0001). The mean hospital stay decreased from 7.7+/-3.6 days in 1991 to 4.1+/-2 days in 1996 (P<0.001). The main biliary complications were related to biliary drainage (2,8%) and retained stones (3.1%). CONCLUSION: This study confirms that laparoscopy is a good alternative with a low complication rate, a short hospital stay, and is an effective and safe option for the management of common bile duct stones. PMID- 10844287 TI - [Role of radical local excision in curative treatment of small low rectal cancers]. PMID- 10844288 TI - [Local excision and adjuvant radiotherapy for rectal adenocarcinoma T1-2 N0]. AB - AIM: To study retrospectively the relapse rate, the functional results and the survival rate in patients with rectal carcinoma treated with local excision and adjuvant radiotherapy. METHODS: Between 1980 and 1995, 43 patients were treated. All cancers were infiltrating tumours except 4 high grade dysplasias with positive margins. The pT classification was: pT1 (n=34), pT2 (n=4), pT3 (n=1). In 4 cases the depth of penetration of the tumor into the bowel wall was not evaluable. The endo-anal excision was performed by surgery (n=20) or by endoscopy (n=23). Only the tumor bed was irradiated in 35 cases (contact x-ray therapy: 30, interstitial iridium implant: 5) and in 8 cases the whole rectum was irradiated with external beam radiotherapy (+/- endocavitary irradiation). RESULTS: Median follow-up was 72 months. Four malignant relapses were observed (local: 1, perirectal lymphatic: 2, distant metastasis: 1). The overall 5- and 10-year survival rates were 80 and 68%, respectively. A total of 41 patients preserved a normal sphincter (95%). The anal function was evaluated as excellent or good in all the cases. No severe radiation toxicity was observed. CONCLUSION: Small rectal carcinomas T1 N0 can be effectively treated in most cases by local excision and postoperative radiation therapy. A close multidisciplinary collaboration is necessary to achieve an optimal result. PMID- 10844289 TI - [Etiological diagnosis of villous atrophy]. PMID- 10844290 TI - [Anti-ulcer agents: indications in adult patients. Recommendations for good practice. French Agency for the Sanitary Safety of Health Products]. PMID- 10844291 TI - [Acute liver failure revealing aorto-caval fistula]. AB - We report the case of a 49-year-old woman with an aorto-caval fistula following lumbar diskectomy. Initial symptoms included isolated hepatalgia, and were later associated with jaundice and hepatomegaly, suggesting acute liver failure due to right heart failure. Only 9 previous cases of postoperative aorto-caval fistula have been reported and our case emphasizes the possibility of a pseudo-digestive presentation of high output arteriovenous fistula. PMID- 10844292 TI - [Fatal subfulminant hepatitis probably due to the combination benazepril hydrochlorothiazide (Briazide)]. PMID- 10844293 TI - [Nephrotic syndrome with acute renal failure due to focal segmental glomerulosclerosis in a patient treated with alpha-interferon for chronic hepatitis C]. PMID- 10844294 TI - [Subfulminant hepatitis E in France]. PMID- 10844295 TI - [Paralytic ileus after treatment with vinorelbine (Navelbine)]. PMID- 10844296 TI - [Duodenal Burkitt lymphoma in an HIV-negative patient: a medical emergency]. PMID- 10844297 TI - [Acute pancreatitis associated with atorvastatine therapy]. PMID- 10844298 TI - [Hemorrhagic anorectal varices: treatment by injection of bioadhesive]. PMID- 10844299 TI - [Apropos of the 'View point' of P. Frileux and R. Parc: 'quality of surgical excision in cancer of the rectum: an important prognostic factor']. PMID- 10844300 TI - [Is the uncinate process resection the key to endonasal dacryocystorhinostomy?]. AB - Endonasal dacryocystorhinostomy is technically more difficult than the classic method and apparently less rational. The nasal fossa is a variable and potentially dangerous anatomic region. The uncinate process "opens the door" to the middle meatus and covers, in almost all cases, the junction between the lacrymal bone and maxillary concha. A marker almost always found. Uncinate process resection is a natural way to access the lacrymal apparatus. Associated with transillumination of the lacrymal duct, uncinate process resection, is in our opinion, a reliable and, most importantly, reproducible method for an endonasal approach to the lacrymal sac. With resection, endonasal dacryocystorhinostomy becomes a simple, safe and rapid procedure. PMID- 10844301 TI - [Evaluation of moderate and severe visual impairments in patients attending an ophthalmology clinic. A prospective study of 1,172 patients]. AB - PURPOSE: Data on blindness and visual impairment in patients attending ophthalmology clinics at the Orleans regional hospital center serving a semi rural area in France were prospectively studied to examine prevalence of visual impairment. MATERIAL AND METHODS: Clinical data on 1,172 consecutive and different patients attending the ophthalmology department during half-days over a 3-month period were collected and analyzed using a computerized system. RESULTS: Prevalence of visual impairment (20/30 or less in the better eye) was 3% in subjects under 60 years of age and rose to 13.7% and 35.9% for patients in the 60 79 age range and over 80 years respectively. For patients under 60, the prevalence of severe visual impairment (20/200 or less in the better eye) was 1.2% and rose dramatically in older patients: 5.8% and 14.6% in the 60-79 year old patients and the over 80 patients respectively. There was no significant difference in the prevalence of visual impairment by sex. Leading causes of severe visual impairment among patients aged 60 years or older were macular degeneration (40%), diabetic retinopathy (16.6%) and cataract (13.3%). The prevalence of irreversible severe visual impairment was 7.4% among patients 60years of age or older. CONCLUSION: A large number of patients attending our ophthalmology clinic have severe visual impairment, particularly those over 60 years of age. Our findings are comparable with extrapolations for population based studies in English speaking countries and can be used to evaluate the prevalence of visual impairment in French rural areas. As the population ages, need for eye health care becomes more and more evident. It is important to organize systematic visual examination in subjects aged 70 to 80 years and to implement educational programs as part of local national health care policies for helping the visually impaired. PMID- 10844302 TI - [The oculomotor effects of the fractures of the floor of the orbit]. AB - PURPOSE: Fractures of the orbit floor have very disabling oculomotor after effects. The objective of this study was to report that close collaboration between the maxillo-facial specialist and ophthalmologist is necessary to avoid oculomotor after effects. METHODS AND PATIENTS: Our study included 10 cases of the fracture of the orbit floor. These cases were recorded between 1992 and 1996. RESULTS: This pathology affected young subjects aged between 20 and 40 years in 70 % of the cases. 80 % of the subjects were males. 90 % of the patients presented a vertical diplopia as a result of a transitory oculomotor deficit. 20 % of the subjects presented a permanent oculomotor deficit at the stage of after effects. All these deficits were of a mechanical origin and, contrary to other authors, we found no deficit with nervous sensory origin. CONCLUSION: Therapeutic success is based on early diagnosis and treatment that requires close collaboration between the maxillo-facial and ophthalmology teams. PMID- 10844303 TI - [Is there a delay in bathing the external eye in the treatment of ammonia eye burns? Comparison of two ophthalmic solutions: physiological serum and Diphoterine]. AB - PURPOSE: An experimental animal study was conducted to analyze the delay for ocular bathing in the treatment of severe ocular ammonia burns. Two solutions of ocular wash, saline solution and Diphoterine were compared. MATERIAL: and methods: The study included 23 eyes of New Zealand albino rabbits that received for 1 minute 100 microl of 15.3% ammonium solution. Each eye was then washed with 250 of saline solution or 250ml Diphoterine after a delay of 1, 3, 5, 10 or 30 minutes. Effects were assessed on the basis of changes in anterior chamber pH, ammonia concentration in the anterior chamber, and cytopathology examination of the burned corneas. RESULTS: Ocular wash with Diphoterine in the first minutes following ocular burn induced an inflexion of the pH curve unlike ocular wash with saline solution. At 30 minutes, there was no inflexion of the pH curve and the ammonia concentration in the anterior chamber was low. Contrary to ocular wash using Diphoterine, stromal edema was seen at cytopathological analysis after washing with saline solution. CONCLUSIONS: This study provides evidence of the interest of ocular bathing in the first minutes following ocular burn by ammonia. The efficacy of external ocular washing with Diphoterine was proven by biochemical and cytopathological demonstrations. The importance of sequelae were related to the degree of initial stromal edema. PMID- 10844304 TI - [Specific causes of blindness and vision impairment in a rural area of Southern Togo]. AB - PURPOSE: To determine prevalences of specific causes of blindness and visual impairment in a rural area of Southern Togo. METHOD: A transversal exhaustive screening was conducted. The sample consisted of the inhabitants aged 5 years and over. From the census, 2293 persons were expected, however, 1738 (75.80%) responded and underwent screening. Each participant had complete ocular examination, and data collected were processed for analysis. RESULTS: The mean age of all participants was 28.02 years, men were predominant. The prevalence of bilateral visual impairment was 2.13%, prevalence of unilateral impairment was 1.15%. Bilateral blindness prevalence was 2.47%, while unilateral blindness prevalence was 2. 65%. Main ocular blinding diseases and their prevalences were as follows: cataracts (8.34%), glaucoma (1.90%), corneal opacities (1. 09%). Women were more affected by bilateral visual impairment, bilateral blindness and cataract.: p= 0.0106 Men were more affected by glaucoma, corneal opacities and unilateral blindness. Participants aged 55 years and over were at high risk of blindness and visual impairment; p= 0.0104. CONCLUSION: This study demonstrated that prevalence of blindness is high, and its major causes are cataract, glaucoma and corneal opacities. A prevention strategy is therefore needed and targeted to these diseases. PMID- 10844305 TI - [Trends and estimates of regional differences of cornea donation and transplantation in France (1991-1998)]. AB - PURPOSE: To evaluate national trends of corneal procurement and transplantation, and to estimate regional disparities between the rates of transplantation and patients registered on local waiting lists. METHODS: A questionnaire was sent, every year since 1994, to all the heads of ophthalmology departments in public and private hospitals. RESULTS: After a dramatic decrease in 1992, donations and transplantations then increased to reach 4549 and 4053 in 1998. This increase was mainly observed for cornea retrieval from non heart beating donors in public hospitals. Since 1997, the number of imported corneas has increased, 1016 corneas were imported in 1998 (25% of the corneas transplanted). The estimate of the number of patients registered on local waiting lists decreased to 7400 at the end of 1998. Regional agreement, using the ratio of the number of patients registered on waiting list at the end of 1998 and the number of cornea harvested in 1998 was low for several French regions: Aquitaine, Champagne-Ardenne, Nord-Pas-de-Calais, Picardie, Ile-de-France, Rhone-Alpes, Antilles. CONCLUSION: Careful thought and regional efforts pointed at non-heart beating donors should be applied to increase the level of corneal harvesting, and to reduce waiting lists in France to levels similar to those in other European countries. Recent issue of legal authorization of cornea banking and creation in october 1999 of a national waiting list should facilitate these efforts. PMID- 10844306 TI - [Macular functions in professional divers]. AB - PURPOSE: Air diving is frequently practiced by professionals or sportsmen. Controversial data exist in the literature on the existence of retinal abnormalities in divers. PATIENTS AND METHODS: 18 divers (aged: 27-58y) who dived around 2000 times in their life were studied: half of them dived only with air while the others used an O2-enriched gas mixture (40 to 60%). None of them had presented a bend (decompression sickness). Visual acuity and ocular fundus examination have been explored. A quantification of color vision and central visual field, so as a fluorescein angiography have been performed. RESULTS: No alteration of visual acuity was noted; abnormalities in the color vision and the visual field are reported; however the angiographic lesions described in the literature have not been observed. DISCUSSION: The alterations of color vision were quite severe but not very frequent. No correlation was found with any characteristics or type of diving. CONCLUSION: These observations are comforting for sportive divers who do not dive very often nor very deep but an individual predisposition is suspected. PMID- 10844307 TI - [Current treatment of retinoblastoma. 153 children treated between 1995 and 1998]. AB - PURPOSE: Treatment of retinoblastoma has changed significantly over the past few years. There are fewer indications for external beam radiation and a new treatment modality, chemotherapy, has appeared. MATERIAL: and methods: We reviewed a series of 153 children treated for retinoblastoma between 1995 and 1998. There were 67 boys and 86 girls: 76 unilateral and 77 bilateral retinoblastomas. Indications for treatments and outcome were reviewed for 230 eyes and for each tumor. Age at diagnosis varied from 0 to 94 months with a median age of 12 months. A family history of retinoblastoma was found in 24 cases. Three children were seen for treatment of recurrence. RESULTS: Among the 76 cases of unilateral retinoblastoma, 56 were enucleated and 20 were treated conservatively (5 with external beam radiation). Among the 154 eyes with bilateral retinoblastoma, 48 were enucleated and 106 were treated conservatively (49 by external beam radiation). Local treatments included chemothermotherapy, laser alone, cryotherapy, and (125)I plaques. Conservative management other than external beam radiation was used for 81 eyes and was successful in 72 (89%). CONCLUSION: We discuss the indications and results of primary chemotherapy and local treatments. External beam radiation is still often indicated in bilaterally advanced cases. In other forms of retinoblastoma, chemothermotherapy is a very reliable and useful treatment. PMID- 10844308 TI - [Place of "exploratory vitrectomy" in the treatment of retinal detachment without proliferative vitreoretinopathy]. AB - INTRODUCTION: One of the principal causes that may contribute to failure in the treatment of retinal detachment without PVR is the inability to detect the retinal break before and during surgery. We propose in these cases the use of exploratory primary vitrectomy allowing the location and the treatment of the retinal break. MATERIAL AND METHODS: We have studied retrospectively 19 cases of retinal detachment without any preoperatively identified break. In 14 cases, it was a pseudophakic detachment (the IOL was in the posterior chamber with an optic between 5 and 6 mm), in 5 cases it was a phakic detachment. Peripheral fundus was examined with the vitrectomy probes with and without perfluorocarbon liquid injection. Cryotherapy or endophotocoagulations have been used to create a chorio retinal adhesion and a gaz tamponade was used without scleral buckling procedure. RESULT: In 2 cases, no retinal break was found. In the other cases, the retinal tear has been identified during basal vitrectomy in 8 cases, during injection of perfluorocarbon in 2 cases and during the vitrectomy done forward the PFLC in 7 cases. The retinal break was identified as a small retinal tear along the posterior margin of the vitreous base in 15 cases (several in 2 cases) and as atrophic hole in 2 cases. DISCUSSION: Exploratory vitrectomy is an interesting technique to identify a retinal break when a scleral indentation cannot offer a good visualization of the anterior retina or retinal tears. In young phakic patients, a primary vitrectomy may be dangerous but seems to be justified in pseudophakic eyes. The research of the tear is sometimes facilitated by a peroperative tamponade of the retro equatorial retina. PMID- 10844309 TI - [Devic's neuromyelitis optica. A childhood case and review of the literature]. AB - Neuromyelitis optical (NMO) or Devic's disease is an uncommon clinical syndrome associating unilateral or bilateral optic neuritis and transverse myelitis within about 8 weeks. Usually reported in adults, childhood cases constitute a distinctive clinical entity with good visual and neurological prognosis without long-term recurrence or sequelae. The pathogenesis is explained by autoimmune demyelinization triggered by a viral disease. Some authors suggest an infectious cause. The CT scan is usually negative while MRI shows nonspecific small foci in the white matter. Unlike multiple sclerosis, the main differential diagnosis, in NMO the cerebrospinal fluid may show pleocytosis with elevated protein level without an oligoclonal band. We report a case of neuromyelitis optica occurring in a 4-year-old child following a flu-like syndrome. Clinical outcome was favorable with a short course of high-dose corticosteroids. PMID- 10844310 TI - [Exophthalmic pneumosinus dilatans: a case report]. AB - Pneumosinus dilatans is a rare disease characterized by progressive development of high pressure in the facial sinuses. The etiology remains unknown. We report a case which developed in the anterior ethmoidal cell and was expressed by exophtalmia. PMID- 10844311 TI - [Birdshot chorioretinopathy associated with tamoxifen retinal toxicity]. AB - We report a case of Birdshot retinochoroidopathy associated with ocular toxicity due to tamoxifen. Adverse drug effects were suspected due to the presence of yellow-white dots in the paramacular region and the fovea and by modifications of the retinal epithelial pigments. Ocular toxicity should be suspected as it may be reversible if recognized and stopped early. Other adverse ocular effects are described and the pathogenic mechanism of tamoxifen retinopathy analyzed. PMID- 10844312 TI - [Round table on bioavailability]. PMID- 10844313 TI - [Ocular drug penetration]. AB - The study of ocular drug penetration may take into account many factors connected to the ophthalmic route specificity: anatomical barriers of the eye, solubility, molecular weight and eyedrops concentration. The factors influencing ocular drug penetration are reviewed. The broad outlines of research focuses now on an increase of the bioavailability, a best tolerability and a reduction of systemic effects of drugs. PMID- 10844314 TI - [Ways to improve ocular bioavailability for topical applications]. AB - The anatomical, physiological and pharmacological properties of the eye explain the short pre-corneal residence time and the poor bioavailability of most eye drop solutions. Many approaches have been proposed to increase ocular bioavailability of drugs. Most eye-drops include a viscosity agent in their formulation to significantly prolong residence time although the increased viscosity is limited due to patient discomfort. More recent developments include biodegradable inserts, eye-drop based on cyclodextrins, liposomes or nanoparticles. PMID- 10844315 TI - [Bioavailability of antibiotics]. AB - To be effective, an antibiotic must achieve therapeutic concentrations at the site of infection. This article is a review on the bioavailability of local and systemic antibiotics. Amongst topical antibiotics, fluoroquinolones and fusidic acid have the best intra-corneal and intra-cameral penetration. Chloramphenicol penetrates the anterior chamber but not always at therepeutic levels. Serum levels are low from topical administration, and haematological toxicity of chloramphenicol eyedrops is still not proven. Amongst systemic antibiotics, the molecules capable of penetrating the eye at therapeutic levels are fosfomycin, imipenem, some of third generation cephalosporins, fluoroquinolones and ureidopenicillins. Intra-ocular penetration of antibiotics is increased by infection, corneal epithelium abrasion, increasing dose frequency or delivery using a biomaterial reservoir (soft contact lenses or collagen schields). Antibiotic-antiinflammatory associations can make infected sites more accessible to antibiotics. The choice of a treatment must take in consideration molecules, dosing and route, and be adapted to the germ to kill and the infected site to treat. PMID- 10844317 TI - [Bioavailability of antiglaucoma drugs]. AB - The biodisponibility of antiglaucoma drugs, generally delivered in an aqueous eye drop solution depends on their capacity to cross the cornea. The structure of the cornea forms a barrier to strongly lipophilic substances and the continuous renewal of the lacrimal film creates a major obstacle, preventing active substances from penetrating the eye. Active substances must thus be delivered in highly concentrated solutions. The systemic bioavailability of antiglaucoma drugs taken orally, for example beta-blockers, is well known, their behavior after eye drop administration remains poorly elucidated and highly dependent on individual susceptibility. We reviewed the literature on pilocarpine, beta-blockers, adrenergic drugs (dipivalyl-epinephrine, apraclonidine, brimonidine), carbon anhydrase inhibitors (acetazolamide, dorzolamide). PMID- 10844316 TI - [Bioavailability of antiinflammatory drugs]. AB - Bioavailability is one of the major factors for therapeutic efficacy. The bioavailability of antiinflammatory drugs depends on the status of the eye (degree of inflammation, vitrectomy, aphakia) and on the drug (route of administration, regimen, concentration and presentation). The bioavailability of steroid drops increases with acetate compounds, frequency of administration, concentration and contact time. Peribulbar injections provide a long-delivery system but with a significant systemic diffusion. Intravitreal steroids have only a local effect due to the very low doses injected. The bioavailability of non steroidal antiinflammatory drugs is variable, but their effects are similar. Antimetabolites such as 5-fluorouracil or mitomycin C have short half-lives, but mitomycin C must be rinsed after use due to its toxicity. PMID- 10844319 TI - ? PMID- 10844318 TI - ? PMID- 10844320 TI - [Residual left orbital phlebolith. An anatomic-clinical observation]. AB - A 29-year-old female patient displayed a dense and hard left orbital mass, inferiorly, 3 years after she was treated through a neurosurgical approach for a left orbital varix. The lesion was hyperdense on CT-scan and was removed through a trans-conjunctival approach, with an excellent result. The histological diagnosis was a phlebolith. PMID- 10844321 TI - [Exposure to light increases the risk of cataracts. Results of the POLA cohort study. Ocular pathologies connected with age]. PMID- 10844322 TI - [Do viagra and fertilization in vitro announce the end of etiologic treatments in andrology?]. AB - Viagra and in vitro fertilization (IVF) with intraovocyte injection of spermatozoa (ICSI) have revolutionized the treatments of impotence and male sterility. They are able to treat successfully most of the cases whatever is the cause of the problem. The andrologist is tempted to renounce to look for an etiological factor and to treat directly his patient. The risk is to miss a diagnosis such as genital tract obstruction, testicular cancer, gonadotropin deficiency, sperm autoimmunity, coital disorders, or reversible toxin exposures, which could benefit from a specific treatment. Moreover IVF can endanger the woman's health and genetic consequences must not be overlooked if ICSI is performed. Concerning impotence a diagnosis of prolactinoma, diabetes or ischemic cardiopathy must not be missed because Viagra can also have cardio-vascular side effects. This article reviews some etiological factors responsible for male infertility or impotence. The importance of a global appraisal of the patient is underlined in order not to limit his role to the one of a sperm producer. PMID- 10844323 TI - [Treatment of ectopic pregnancy in 2000]. AB - Operative laparoscopy is currently the best treatment for pregnancy (EP). As with laparotomy, laparoscopic treatment of EP can be either conservative (salpingotomy or radical (salpingectomy). After conservative laparoscopic treatment, failures are diagnosed by monitoring the drop in beta-hCG levels. Fertility results after laparoscopic treatment of EP are comparable with those observed after similar treatment by laparotomy. Better knowledge o the risk factors of EP, development of hCG assays using serum progesterone and high resolution sonography using vaginal probes allow early diagnosis of EP and a nonsurgical approach in more than 30% of cases. When inclusion criteria are strict, methotrexate administered by local injection or systemically (1mg/kg) in a single dose or in combination with mifepristone gives a 90 to 95% success rate. Whatever treatment protocol is used, fertility prognosis after EP is not correlated to the features of EP but depends mainly on patient age and past history. PMID- 10844324 TI - [Hyperhomocysteinemia and pregnancy: a dangerous association]. AB - Homocysteine results from the demethylation of the essential amino acid methionine. Its metabolism depends primarily on three enzymes and several vitamin cofactors (vit. B6, B9 and B12). Genetic abnormality in these enzymes or deficiency of these vitamins lead to Hyperhomocysteinemia. Hyperhomocysteinemia belongs among the congenital thrombophilies and is a long-known vascular disease risk factor. The discovery that hyperhomocysteinemia may also be responsible for several pregnancy complications has only recently been made. Studies in this area are still scarce and report on limited numbers of patients. It nevertheless appears clear that HHCh is associated with the syndromes of repeated miscarriage, pre-eclampsia, placenta abruptio, thromboembolic events, neural tube defects, and perhaps with fetal death-in-utero and intra-uterine growth retardation. Supplementation with vitamin B9 can reduce plasma HC levels, and is thus recommended in patients with HHCh. The prevention of thromboembolic events during pregnancy by anticoagulant treatment is also desirable in these patients. PMID- 10844325 TI - [Grand prematurity, risk of neuropsychic handicaps and neuroprotection]. AB - Increased survival of very preterm infants and extremely preterm infants does not imply an increase in neurodevelopmental sequelae. However, preterm infants remain at high risk of severe cerebral palsy with neuromotor dysfunction and mental retardation. Necrotic or hemorrhagic lesions occurring in the periventricular ring of telencephalic white matter are the most threatening events for the developing brain of these infants. Recent progress in neuroepidemiology, developmental neurobiology and imaging methods has made it possible to revisit the pathophysiology of brain lesions on a multifactorial basis. Treatment and early detection of infections is a priority. Neuroprotective agents capable of arresting neuron cell death (antagonists of the excitotoxic cascade, free-radical antagonists, proinflammatory cytokine antagonists, growth factors) are new strategies for the prevention of cerebral palsy. PMID- 10844326 TI - [Antenatal diagnosis of limb body wall complex]. AB - The "limb body wall complex" or LBWC is a rare polymalformative syndrome. Two distinct phenotypes of which were recently described: one form with "placento cranial" adhesion and the other with "placento-abdominal" adhesion. Coelosomia is found in all cases, it variably coexists with encephalic, vertebral, visceral or limb anomalies. Three pathogenesis are proposed: the "early amnion rupture" theory and the "vascular" theory can explain the form with "placento-cranial" adhesion, conversely, the "defective folding process" allows for a better explanation of the form with "placento-cranial" adhesion. These theories do not exclude each other if we accept that this syndrome could be split: the first form would then be the original "LBWC" et could belong to "amniotic band syndrome", the mechanism of which is not precisely known, whereas the second could be named "body stalk syndrome" - "syndrome du cordon court". In any case, this syndrome should be better known so as to be studied and give a prognosis for a later pregnancy, because there is no recurrence. PMID- 10844327 TI - [Vaginal route treatment for residual urinary incontinence after closing an obstetrical fistula: apropos of 49 cases]. AB - OBJECTIVE: Described here is a simple treatment for female urinary incontinence present and concurrent cystocele repair, if present. The objective of this technique which combines a bladder neck and a suprapubic percutaneous suspension, is to obtain by a sling effect, efficient cure of the urinary incontinence, in avoiding the common retention problem. PATIENTS AND METHODS: Between 1992 and 1998, 49 black female patients were treated with this procedure. Average age was 18 years old (the youngest 14, eldest 29 years old) 39 were primigravid while 10 were multigravid (2 to 6 childbirths). 12 (24. 4%) have undergone at least 2 operations for fistula cure, 6 of the patients showed previous incontinence surgery; Burch technique (n=1), Martius fat sling (n=3) and anterior colporraphy (n=2). 25 patients (51%) were suffering from a severe incontinence, for 17 cases the urine leak was grade 2 and 8 cases grade 3 at decubitus. RESULTS: No bladder lesion occurred on any of the 49 patients. Except for one general peritonitis which occurred on a patient who was receiving a combined recto-vaginal fistula operation, the post operative complications were minor (n =7). We observed only one direct recurrent urinary incontinence which was treated a second time with the same technique and was cured. For 46 studied cases and an average follow-up of 28.6 months, 29 patients (63%) were declared totally cured, 8 (17.3%) showed functional improvement, 9 (19.5%) remained incontinent. The global success rate (cure and improvement) was obtained in 80% of cases. The results were less satisfactory in multi-operated patients, particularly in those who suffered from extensive cervico urethral damage (Group III). In this group only one out of two patients obtained cure or improvement. CONCLUSION: Due to its real simplicity the bladder neck and vagina wall sling is easy to redo, and because its reliability we have chosen this procedure as a choice treatment for incontinence following a vesico vaginal fistula. The excellent results confirmed by a longer follow-up has led us to extend its indication to all types of female urinary incontinence. This technique is particularly well suited for use in developing countries. PMID- 10844328 TI - [Gestational diabetes: the Rennes Study of joint obstetrical and endocrinological management]. AB - OBJECTIVE: To assess the efficacy of a co-program of care in gestational diabetes conducted by the obstetrical and endocrinological units of the Rennes South Hospital (Hopital Sud) and the possibility of adapting obstetrical care to existing risk factors. PATIENTS AND METHODS: A retrospective analysis was made of 200 cases of gestational diabetes diagnosed between January 1993 and December 1996 in the obstetrical units. RESULTS: Instrumental extraction and cesarean section were required for 13.5% and 20.5% of the deliveries respectively. Shoulder dystocia occurred in 2%. 19.9% of the infants had macrosomia and 5. 3% were small for gestational age. Neonatal morbidity required transfer to the pediatric intensive care unit for only 2.9% of the infants. Frequency of cesarean section and macrosomia correlated with poor blood glucose control and excess maternal weight for gravidic hypertension. At 3 months post-partum, glucose intolerance and non-insulin-dependent diabetes were diagnosed in 13% and 2% respectively. CONCLUSION: Systemic screening and obstetrical and endocrinological care allowed patients to prevent maternal and fetal complications in gestational diabetes and to initiate hygiene and dietary habits for the prevention of post partum non-insulin-dependent diabetes. Obstetrical care can be adapted to risk factors such as overweight, late screening or poor blood glucose control. PMID- 10844329 TI - [Delivery of large baby after cesarean section: role of trial of labor. Apropos of 355 cases]. AB - OBJECTIVE: Our goal is to determine whether a trial of labor in women with suspected fetal macrosomia would be a valuable alternative to elective repeat cesarean. MATERIAL: and methods: Based on retrospective analysis of 355 women with previous cesarean section who delivered macrosomic infants (> or =4,000g), we tried to determine the impact of fetal weight on a trial of labor. The outcomes of trial of labor with fetal macrosomia were compared on the one hand to those of elective repeat cesarean and on the other hand to those of trial of labor with normal birth weight (<4,000g). RESULTS: The trial of labor was conducted in 297 cases (83,7%), and had led to vaginal birth in 189 cases (63,6%). There were 4 uterine ruptures (1,3%) and 8 uterine dehiscences (2,7%) among the women who underwent a trial of labor. In this group, there were 4 perinatal deaths (1,3%) related in one case to uterine rupture, and 2 brachial plexus injuries related to shoulder dystocia after vaginal birth. Perinatal and maternal outcomes of trial of labor were similar to those of elective repeat cesarean. A trial of labor was more associated with scar separations and lower success rate if the infant weighed 4,000g or more. CONCLUSIONS: It appears that the use of trial of labor for delivery of large baby with prior cesarean section was associated with lower success rate and the maternal and fetal risks could be increased. However, carefully others controlled studies are necessary to establish the appropriate management in this setting. PMID- 10844330 TI - [Maternal transport to the Bordeaux University Hospital: a retrospective study of 263 cases (1996-1998)]. AB - OBJECTIVES: To describe a population of high risk pregnancies transported to a regional perinatal care centre (level III) and to analyze the advantages and limitations of the organization of this perinatal care network. METHODS: Retrospective study of 263 patients transported to the University Hospital of Bordeaux between September 1996 and September 1998. RESULTS: Maternal transport mainly came from the Gironde department (53%) and from level I care hospitals (66%). The rate of multiple pregnancies was 17%. Principal indications were preterm labor with or without rupture of fetal membranes (52%) and preeclampsia with or without intrauterine growth restriction (28%). The rate of returns to the referring care centre was 2%. The mean term at delivery was 32 WA. Forty-one percent of patients delivered within 24 hours of transport. The rate of caesarean sections was 55%. There were 303 live births. The mean duration of hospitalization of the neonates was 28 days. The rate of perinatal mortality was 6.2%. Among the 96 pregnancies beyond 32 WA, 17 (6.5%) could have been transported to some level IIb care hospitals, nearby the patient's residence, without changing maternal and neonatal prognosis. Neonatal mortality was not significantly different for maternel transport (18%) or neonatal transport (14%) in the group of premature infants born prior to 31 WA in our study. CONCLUSION: Our organization is currently trying to improve transport management (non emergency situations, collaboration of level II care hospitals) and to increase the rate of returns to the referring care centres. PMID- 10844331 TI - [Uretero-uterine fistula: what is the role of ureteroscopy?]. AB - We present a case of uretero-uterine fistula that occurred following cesarean section complicated by postoperative peritonitis subsequent to failure of the uterine sutures. The distal and the proximal ends of the ureter were successfully explored by ureteroscopy. We discuss the contribution of ureteroscopy in ureter exploration and endoscopic treatment of uretero-uterine communications. PMID- 10844332 TI - [What is the current training of the gynecology-obstetric intern in France?]. PMID- 10844333 TI - ? PMID- 10844334 TI - [Multi-slice helical CT: biggest pitch does not mean dosimetric bargain]. PMID- 10844335 TI - [Why quality management is imposed in radiology?]. AB - Quality management plays an important role in the French health care system, especially in radiology. The purpose of this article is to review the reasons of this evolution. Radiologists, technicians and hospital managers must be familiar with these concepts and implement them in their units. PMID- 10844336 TI - [Macroscopic intrahepatic portosystemic venous shunt: review of the literature and reclassification]. AB - Macroscopic intrahepatic portosystemic venous shunts (IHPSS) are defined as communications between the portal and the systemic venous circulation, measuring more than 1mm in diameter, and at least partially located inside the liver. Four different types can be identified based on anatomical, clinical, and pathophysiological criteria. Type I includes patent paraumbilical veins, located in the liver, and commonly encountered in portal hypertension. Types II, III, and IV are much less common and have been reported in only 47 publications in the entire French and English literature. They include shunts, unique or multiple, between a portal branch and a hepatic vein, located either in two adjacent liver segments (type II) or in non-adjacent liver segments (type III). Type IV corresponds to any tubular communication developed between the right portal branch and the inferior vena cava. The exceptional patent ductus venosus or a patent umbilical vein should not be considered as IHPSS since their course is strictly extrahepatic. PMID- 10844337 TI - [Therapeutic ligature of hypogastric arteries: color Doppler follow-up]. AB - PURPOSE: To evaluate the interest of color Doppler sonography in the watching of therapeutic ligature of hypogastric arteries. PATIENTS AND METHODS: Thirteen female patients presenting a serious haemorrhage in the post-partum and be in managed by bilateral hypogastric ligature, have undergone a systematic protocol of investigation involving a Doppler sonography in the fourth day after intervention and then monthly until repermeabilization of the internal iliac arteries. RESULTS: In all cases, the first exam showed down-side the ligature, a flow inversion with an important pelvic collateral circulation. The next Doppler exams showed repermeabilization of the hypogastric arteries after an average interval of 5 months (extremes: 1 to 6 months). CONCLUSION: Color Doppler showed the resolvent character of therapeutic ligature of the hypogastric arteries and may estimate with sufficient accuracy the interval of repermeabilization. PMID- 10844338 TI - [HASTE sequence and cine-MRI evaluation of the cervical spinal canal: evaluation in 11 healthy subjects]. AB - PURPOSE: We studied 11 healthy subjects were evaluated using cine MR imaging comparing HASTE and gradient echo sequences. Materials and methods :HASTE is a high-speed turbo-spin echo T2-weighted sequence. All examinations were performed using dynamic MRI using a simple process allowing acquisition of images at different stages during flexion and extension. The cine MR evaluation was obtained by rebuilding a cine-loop sequence. RESULTS: HASTE sequence provides a myelographic effect of the cervical spine. The size, the pattern and the movements of the cervical spine, the spinal cord, as well as the functional reserve of cerebro-spinal fluid are analysable. CONCLUSION: Because of its very high speed, HASTE sequence is particularly useful for cine MR imaging. PMID- 10844339 TI - [CT enteroclysis for detection of small bowel tumors]. AB - PURPOSE: To assess the feasibility and the usefulness of CT enteroclysis (helical CT with enteroclysis) in detecting small bowel tumors. MATERIALS AND METHODS: Fifty patients were referred for suspicion of small bowel tumor. CT enteroclysis is performed by injecting a large volume of water using a pomp through a nasojejunal tube followed by a thin section helical acquisition. RESULTS: Forty eight helical CT enteroclysis were performed in order to detect 25 small bowel tumors. Among them 22 were confirmed by histological study. The mean size of tumors was 23 mm. In 12 of 17 cases, diagnosis was missed or incomplete at conventional barium study. Enteroscopy was performed in 12 of 22 cases, with discordant result in one case and incomplete result in 3 cases. In 8 cases, including 5 carcinoid tumors, patients had surgery after CT enteroclysis only, enteroscopy would probably have not made the diagnosis because the lesions were far from the duodenojejunal junction and ileocaecal valve. CONCLUSION: Helical CT enteroclysis is a new method for detecting small bowel tumors, easy to perform, well tolerated. It seems to be more sensitive than conventional barium studies and less invasive than enteroscopy. Tumor characterization and staging can be performed using a single examination. It seems to be justified to perform CT enteroclysis to detect small bowel tumors or in the evaluation of patients with polyposis. PMID- 10844340 TI - [Pelvic ganglioneuroma. Case report]. AB - Ganglioneuroma is a neurogenic tumor originating from autonomic ganglia. It most commonly occurs in patients younger than 20 years with a slight predominance of females. The pelvic localization is rare. Imaging can lead to diagnosis but it is not specific. The diagnosis can only be confirmed by the pathologic study. The authors report one case of pelvic ganglioneuroma diagnosed in an 11-year-old child who also had hypertension and renal failure. PMID- 10844341 TI - [Leiomyosarcoma of the superior vena cava: diagnosis by endovascular biopsy]. AB - We report the case of a 69-year-old woman with leiomyosarcoma of the superior vena cava presenting with acute superior vena cava syndrome (SVCS). CT and MRI failed to fully characterize the endovascular process. Percutaneous endovascular biopsy, followed by metallic stent placement to treat the SVCS, confirmed the diagnosis. Symptoms resolved within 48 hours and surgical resection of the tumor was performed one month later. Unfortunately the patient died two weeks later because of intracranial hemorrhage. PMID- 10844343 TI - [Solid pleural-based mass as initial manifestation of non-Hodgkin's lymphoma]. AB - Subpleural involvement is a well-known site of advanced or recurrent lymphoma. But, to our knowledge, it has never been described as an initial manifestation of this disease. We report the case of a patient with solid subpleural mass revealing a non-Hodgkin's lymphoma. PMID- 10844344 TI - [What is it? Abdominal foreign body and small intestine fistula]. PMID- 10844342 TI - [Pancreatic pseudocyst: unusual presentation of polyarteritis nodosa]. AB - Even though pancreatic involvement in case of polyarteritis nodosa is uncommon, it reveals infrequently the disease. Furthermore, the occurrence of pancreatic pseudocyst in case of polyarteritis nodosa is rare. One case of intrapancreatic pseudocyst complicating chronic pancreatic during polyarteritis nodosa is reported. CT scan showed non specific intrapancreatic cystic lesion for which the diagnosis of mucinous cystic tumor of the pancreas was considered. After surgical resection, histopathologic analysis led to the diagnosis of intrapancreatic pseudocyst secondary to polyarteritis nodosa. PMID- 10844346 TI - Introduction PMID- 10844345 TI - [NAAEH recommendations. Management of severe cranial injuries in the early phase. Text of the recommendations. National Agency of Accreditation and Evaluation in Health]. PMID- 10844347 TI - [Body changes during pregnancy]. AB - Pregnancy induces numerous changes in the physiology of the woman. Those changes are necessary for the embryo and fetus to have a normal growth, and for the woman to adapt to that physiologic event. A 50% raised cardiac flow is the consequence of increased systolic flow and cardiac frequency. Blood volume is about 40% larger than in non-pregnant woman. It is the consequence of enlargement of the plasmatic volume (50%), and the red cells mass (30%). Those different changes explain the physiologic anemia of pregnancy. The main part of the blood volume increase corresponds to an enlargement of the venous system, but without any change of the central venous pressure. Arterial pressure remains unchanged throughout the pregnancy, or sometimes gently decreases (10%). Those changes are more important during an effort, particularly during the labor. Increase of cerebral blood flow (as a consequence of a raised cardiac flow) is limited by the cerebral autoregulation. As a consequence, there is no evidence for dramatic cerebral hemodynamic changes during pregnancy. Nevertheless, autoregulation is less effective for arterial pressure over than 150 mmHg, what can induce an hemorrhagic stroke. Blood levels of steroid (progesterone, oestrogens) and peptidic (HCG, HPL) hormones are increased. Oestrogens are said to make capillary fragile, and progesterone is responsible for the enlargement of the venous system. In order to prevent an hemorrhagic accident, pregnancy induces a lack of fibrinolysis, and an excess in coagulation. The consequence is the ability of the pregnant and post-partum woman to develop venous thrombosis. PMID- 10844348 TI - [Tumors of the neuraxis and neuraxial sheaths during pregnancy (excluding tumors of the sellar region)]. AB - An association between central nervous system neoplasm and pregnancy is uncommon although its frequency is underestimated. The most commonly observed tumors are acoustic neuromas, meningiomas and gliomas. The interactions between these tumors and pregnancy is not well known : Pregnancy could affect tumor growth by : - activating tumor growth by stimulating receptors; - increasing the peritumoral edema; - favoring immunotolerance due to pregnancy. Steroid hormone receptors and their therapeutic significance are described here, focusing particularly in meningiomas. Central nervous system tumors do not effect the progress of pregnancy per se but do the therapeutic implications. The clinical aspects are the same as in non-pregnant women. MRI appears to be the most adapted imaging technique. Neurosurgical guidelines are also described, with emphasis on symptomatic therapeutics, especially for seizure. Attitudes concerning the nature of the tumor and time of pregnancy are then discussed. The obstetrical attitude should be therapeutic abortion or delivery, with cesarean if necessary. Prognosis is getting better for the child and mother alike, but still remains poor. PMID- 10844349 TI - [Pituitary disorders in pregnancy]. AB - During pregnancy there is a normal increase in the volume of the anterior pituitary as demonstrated by MRI and hormone secretions which increase (PRL) or decrease (FSH, LH). During pregnancy pituitary adenomas, especially prolactinomas, may evolve as in non-pregnant women (microadenomas) or differently (macroadenomas). In 35 % of cases macroprolactinomas worsen during pregnancy making their medico-surgical management mandatory prior to pregnancy. Inversely, pregnancy occurring in a subject with a microprolactinoma never induces severe local complications so such tumors may be managed surgically or medically. Surgery should be performed for acromegaly or Cushing's disease before or early in pregnancy. Subacute pituitary apoplexy (intratumoral hemorrhage) occurs in about 10 to 15 % of adenomas but, generally speaking, clinical symptoms remain mild in pregnant women. Lymphocytic hypophysitis occurs at the end of pregnancy, or during the post-partum period. The association of complete pan-hypopituitarism and hypersignal on MRI examination may mimic hypophyseal apoplexy and could lead to and inappropriate surgical procedure. PMID- 10844350 TI - [Cerebro-meningeal hemorrhage secondary to ruptured vascular malformation during pregnancy and post-partum]. AB - Thirty two new cases of vascular malformations discovered during pregnancy were reviewed in a multicentric retrospective study and compared to the literature. The authors studied both the influence of pregnancy and post-partum on cerebral vascular malformations and the corresponding neurosurgical and obstetrical management. The relative frequencies of arteriovenous malformations (AVMs) and arterial aneurysms (AAs) is the same for pregnant and non-pregnant women. Pregnancy does not increase the risk of first bleeding of vascular malformations but the risk of re-bleeding is increased for AVMs. The bleeding of a vascular malformation is more strongly correlated to age than to parity. The mean maternal age at the time of rupture is greater for AAs than for AVMs, as it is in the general population. The bleeding of a vascular malformations occurs more frequently during the second and third trimesters of pregnancy than in the first one and in the post-partum. Labor and delivery are not great-risk-periods. Management of vascular malformation in pregnant woman is proposed as follow : --a ruptured AA or AVM is managed as it is in non-pregnant woman, whatever the gestational age; no fetal extraction needs to be performed except ed if the rupture occurs at the very end of the pregnancy;--an non-ruptured AVM discovered during pregnancy is treated after delivery; an non-ruptured AVM that never bled before pregnancy is managed in the same way that an non-ruptured AA. PMID- 10844351 TI - [Vascular cerebral thrombosis during pregnancy and post-partum]. AB - Pregnancy and puerperium are associated with a number of cerebrovascular conditions that may result in stroke. Those include cerebral venous thrombosis and cerebral arteries occlusions. Comparing stroke rates during pregnancy with those of non-pregnant women showed only a marginal excess risk during pregnancy and puerperium. Strokes due to cerebral venous thrombosis represent 10-20 per 100 000 deliveries in western countries. The cause of intracranial venous thrombosis is usually unknown. However, better understanding of abnormalities in coagulation leading to intravascular clotting in the early puerperium is resulting in better understanding of this disease. Nevertheless, an etiological work up should be performed, particularly when the thrombosis occurs during pregnancy. Its clinical manifestations often include focal neurological signs, seizures and headache. Alterations in consciousness occur as intracranial pressure increases. Arterial occlusions account for about 60% to 80% of cerebral ischemic lesions. A probable cause of ischemic stroke is diagnosed on the basis of clinical, biological and radiological data. Eclampsia is the main cause of nonhemorrhagic stroke. A search for rare causes of stroke linked to pregnancy such as post-partum cardiomyopathy, paradoxical embolism, choriocarcinoma, cardiac and hematological disorders may be appropriate. PMID- 10844352 TI - [Head injury and pregnancy]. AB - Even mild, head injury during pregnancy can threaten either the maternal or the fetal life. Traffic accidents are the main causes so head injury is often associated with other lesions, frequently abdominal trauma. Among all these possible lesions, head injury and hemorrhagic shock are the main sources of fatality in pregnant women. All kinds of trauma and especially head injuries have a potentially deleterious effect on fetal viability. This risk is associated with systemic and cerebral consequences of post-traumatic arterial hypotension, anoxia or anemia. Moreover, diagnostic procedures and medications can add their noxious secondary effects, contributing more or less to poor fetal outcome. Decision making is necessarily dictated by the severity of the consciousness disorders and/or the encephalic lesions. A convergent discussion between the intensive care physician, the neurosurgeon, the obstetrician and the anesthesiologist is needed. Severe or prolonged traumatic coma is highly dramatic situation because the maternal outcome remains largely unknown for days or weeks. Technically, for these severe comas, there are no substantial differences with what is usually done in a similar case without pregnancy. Neurosurgical indications follow the usual rules, except that some non-urgent indications should be delayed for a while. Usually, this simple rule of independency in indications also applies to the obstetrician. Special care must be taken for fetal monitoring required throughout the initial critical phase. PMID- 10844353 TI - [Hydrocephalus during pregnancy with or without neurosurgical history in childhood. Practical advice for management]. AB - Many women with cerebrospinal fluid shunts are now reaching reproductive age. Clinical management of pregnant patients with hydrocephalus should include preconception counseling and CT scan or MRI. A family pedigree should also be established for counseling on the risk of recurrence of the woman's condition or another neural tube defect. Electrophoresis of acetylcholinesterase in the amniotic fluid can provide the diagnosis of open neural tube defect between 13 and 24 weeks gestation. Shunt malfunction may occur during pregnancy in 50 % of cases. Management requires well-planned, a combined neurosurgical and obstetrical approach. Vaginal delivery is possible in asymptomatic mothers. Cesarean section is recommended for neurologically unstable patients. Prophylactic antibiotics are recommended for labor and delivery to avoid shunt infection. Epidural analgesia is contraindicated in patients with intracranial hypertension. Some complications of complementary treatment for cerebral tumors in childhood are briefly reported. PMID- 10844354 TI - [Comparative study of fixation mode in total knee arthroplasty with preservation of the posterior cruciate ligament]. AB - PURPOSE OF THE STUDY: We performed a prospective randomized study to compare two fixation modes, with and without cement, for total knee arthroplasty. MATERIAL AND METHODS: The study series was composed of 96 cemented or noncemented total knee arthroplasties performed between May 1993 and October 1995. The only difference was the diamond interface used for cemented prostheses and the mesh interface used for uncemented prostheses. The operator was unaware of the type of fixation until the bone slices had been obtained. We assessed outcome in 73 cases with a mean follow-up of 27 months. The two populations were comparable for preoperative clinical status, bone tophicity and surgical procedure. RESULTS: The mean duration of the operation was sgnificantly longer (> 10 min) for the cemented protheses. The complication rates were comparable but we did have two mobilizations of the tibial implant in the noncemented group. The total scores (127 +/- 29 in the cemented group versus 135 +/- 20 in the uncemented group) were significantly different. There were however more cases with degradation of the controlateral knee in the cemented group although the difference was not significant. When these cases were excluded from the analysis, the total scores for two groups were similar (143 and 140 respectively). Radiographic outcome was quite different with mobilization of the tibial implant in 2 cases and the rate of lucent borders was significantly higher in the noncemented group. DISCUSSION AND CONCLUSION: While the clinical outcome was comparable, the quality of the fixation was significantly better with cemented arthroplasty, which remains the gold standard. PMID- 10844355 TI - [AT-TG (anterior tuberosity-trochlear groove): interobserver variability in CT measurements in subjects with patellar instability]. AB - PURPOSE OF THE STUDY: Measurements between the anterior tuberosity of the tibia and the trochlear groove of the patellar surface of the femur AT-AG) are recognized parameters for exploring patellar instability. Interobserver variability should be less than 4 mm and reproducibility should be greater than 85 per cent for valid measurements. We assessed interobserver variability with CT measurements. MATERIAL AND METHOD: The radiography files (left and right knee) of thirteen patients with patellar instability were examined by six radiologists working in private practice after training in our university center. RESULTS: The results demonstrated major variability with acceptable reproducibility in only 15% of the measurements. Differences greater than 5 mm were observed for 14 groups of values (54 per cent) and greaterd than 8 mm in 5 groups (19 per cent). The spread was mainly related to determinations of the TG point in case of major dysplasia and in defining the precise contour of the tibial tuberosity. Surgical decisions made on the basis of these millimetric values should be made prudently. PMID- 10844356 TI - [In vitro effects of growth hormone on osteoclastic activity: clinical applications]. AB - PURPOSE OF THE STUDY: This study was designed to investigate the in vitro effects of human growth hormone (hGH) on osteoclastic resorption in a nonfractionated rabbit bone cell model. MATERIAL AND METHODS: Rabbit bone cells were cultured on dentine slices in the presence of parathyroid hormone and vitamin D3. The percentage of dentine slice surface resorbed, number of lacunae per surface unit and mean area of lacunae were compared between cell cultures grown in the presence of graded concentrations of hGH and human insulin-like growth factor-1 (hIGF-1) and controls. RESULTS: After 4 days of culture, rabbit bone cells cultured on dentine slices in the presence of hGH and hIGF-1 showed significantly stimulated osteoclastic resorption activity. When neutralizing anti-hIGF-1 anti serum (4 microg/l) was added to the starting culture, the stimulatory effects of hIGF-1 and hGH on osteoclastic resorption activity were totally abolished. DISCUSSION: These findings indicate that the effects of hGH stimulation on osteoclastic resorption in vitro are mediated via local hIGF-1 secretion by stromal cells such as osteoblasts. Proteases appear to play a role in the degradation of the organic matrix. Our experiments show that hIGF-1 and hGH stimulate the production of matrix metalloproteinases MMP-9 and MMP-2. Similar to the resorption activity, hGH stimulates protease activity via stromal cell production of hIGF-1. CONCLUSION: This study suggests that natural or synthetic MMP inhibitor modulation of protease activity could reduce the degradation of the organic matrix and then prevent, for example, inflammatory reactions subsequent to prosthetic loosening. PMID- 10844357 TI - [Hand on composite prosthesis after resection of peri-acetabular sarcoma]. AB - PURPOSE OF THE STUDY: To improve function after pelvic resection involving theacetabulum we have used an anatomic composite implant built with screws and cement. MATERIAL AND METHOD: Since 1990, 27 patients with peri-acetabular bone sarcoma have been treated by extensive resection followed by "hand on innominate prosthesis with constrained total hip prosthesis. The hand on innominate prosthesis was made of a titanium cup, a set of long titanium screws and 2 or 3 packs of gentamycin loaded ciment. RESULTS: Many postoperative complications were observed; deep infection (15 p. 100), hip prosthesis dislocation (33 p. 100) and local recurrence (17 p. 100). Nine patients (33 p. 100) had to be reoperated. Nevertheless at last follow up 24 patients still had their composite prosthesis. The mean functional result, rated according to a modified Enneking's staging system, was 83 p. 100 with unlimited walking without support, average hip flexion 100 degrees, length discrepancy less than 1 centimeters. DISCUSSION: These results were similar to those described in the literature for custom made innominate prosthesis and much better than those of alternative reconstructive procedures. Hand on composite prosthesis is cheaper, easier, more adaptable and enables better anchorage than custom-made prosthesis. Such a procedure can be used even after total iliac wing resection. CONCLUSIONS: The advantages of such procedure pleads for using it extensively after acetabular resection. But long term follow up is necessary to validate indications. PMID- 10844358 TI - [Proximal femoral reconstruction with megaprosthesis versus allograft prosthesis composite. A comparative study of functional results, complications and longevity in 41 cases]. AB - PURPOSE OF THE STUDY: To compare femoral reconstruction using megaprosthesis versus allograft prosthesis composite. MATERIAL AND METHODS: Forty-one consecutive proximal femoral reconstructions with an allograft-prosthesis composite (21 cases) or a megaprosthesis (20 cases) after tumor resection were reviewed in a retrospective study. The following criteria were considered: functional outcome; long term survival; complications. Chi-square test and Wilcox tests were used to compare groups. The medium and long-term survival curves for these reconstructions were made using the Kaplan-Meier standard methods. The failure of prosthesis was defined as revision for mechanical failure (either aseptic loosening or dislocation), for infection or local recurrence. The comparison of the curves was performed using the Log-Rank test. RESULTS: Infection (10 p. 100) and instability, in both groups, and loosening, in the megaprosthesis group, were the common causes of failure. There was difference between functional results in the two groups (limping and crutches using was more lower in allograft-prosthesis composite group). Survival analysis showed a 5 and 10-year survival of 77 +/- 12 p. 100 for the patients with composites. Five and ten - year survival were 73 +/- 11 p. 100 and 0 p 100 respectevely for those with megaprostheses. No significant difference was noted between survival of these two groups but a tendancy (p =0.09). Radiological allograft resorption was noted for more than 50 p. 100 of allograft composite prosthesis without modification of functional result or symptomatic loosening. DISCUSSION: The functional results seem better in the composite group when compared to the megaprosthesis group. Reconstruction of the abductor mechanism is essential to stabilize the prosthesis and to decrease the limp. When the great trochanter cannot be preserved, we used suture of gluteus medius tendon to tensor of fascia lata, which is re-enforced using a piece of biceps femoris. The dislocation rate was approximately the same in our two groups. Several authors reported a lower dislocation rate with composite reconstructions than massive prosthesis. The rate of infection is similar to other reported series. In our study it has been possible to show a tendancy for superior survival of the composite reconstruction. When the review was later than 5 years the radiological appearance of the graft in our series was often concerning with resorption or fragmentation present in six of the eight cases. This radiological appearance is not as yet responsible for any revision or any change in the functional result however it does remain a worry. CONCLUSION: Composite reconstructions probably allow a better functional result when considering proximal reconstruction of the femur. The radiological appearance of these allografts in the long term is however worry some without any evidence so far of worsening functional level or any evidence of prosthetic loosening. It would seem to us that the current level of knowledge would advocate the use of massive allografts together with prosthesis. This does seem still to remain the best choice for proximal femoral reconstruction. PMID- 10844359 TI - [Tumoral calcinosis of the foot in patient on hemodialysis]. AB - We report a new case of tumoral calcinosis in a renal failure patient. In this patient the pseudotumoral formation was found in an unusual localization on the medial border of the foot. The condition progressed with successive development of bilateral tumors and septic contamination. Pathology confirmed the diagnosis. The patient was successfully treated with tumor resection and renal graft. The origin of these pseudotumors remains a question of debate. Several pathogenc mechanisms have been put forward. Pronosis is always favorable. Several authors have suggested a phosphorus calcium work-up can be useful for classifying these tumors, guiding surgical treatment if required and managing the underlying cause. PMID- 10844360 TI - [Primary intraosseous rhabdomyosarcoma]. AB - We report a case of primary intraosseous pleiomorphic rhabdomyosarcoma located in the pelvis of a 21-year-old woman followed for 4 years. The lytic tumor involved the acetabulum and the isthma with moderate extension to soft tissue. First line chemotherapy was unable to arrest tumor progression. Hemipelvectomy with saddle prosthesis reconstruction was performed, but septic complications dictated a secondary inter-ilio-abdominal amputation. Recurrence-free remission was achieved for 4 years, suggesting this was indeed a primary tumor. Primary intraosseous rhabdomyosarcomas are exceptional. Bone localizations generally suggest metastasis from a primary tumor often situated in an intraperitoneal localization. When search for a primary tumor is negative, intraosseous lesions can be considered as primary tumors warranting curative treatment. Radical surgical resection is recommended within the framework of a multidiscipinary management protocol associating radiotherapy and chemotherapy to improve prognosis. PMID- 10844361 TI - [Scoliosis and diastematomyelia: four cases and a review of the literature]. AB - We report four cases of scoliosis associated with diatematomyelia observed between 1984 and 1998. The patients were four girls aged 10 to 12 years. Skin lesions were found on the midline in 3 cases and 2 had a neurological disorder. A myeloscan was performed in two cases, tomomyelography in one and MRI in one. Three patients were operated on with good outcome. The fourth child is under orthopedic treatment. We reviewed the literature on scoliosis with diastematomyelia. PMID- 10844362 TI - [Isolated palsy of the flexor pollicis longus in anterior interosseous nerve syndrome]. AB - Anterior interosseous nerve palsy are relatively rare, specialy in traumatic cases. Two case of incomplete anterior interosseous nerve palsy with isolated total loss of function of flexor pollicis longus are reported. Partial palsy of the A.I.N. have been previously reported, but their mechanism is unclear. The incomplete anterior interosseous nerve palsy has to be known. They can be mistaken with tendon rupture, specialy in traumatic cases, and lead to unnecessary surgery. PMID- 10844363 TI - [Henri horoszowski] [In Process Citation] PMID- 10844364 TI - [Intracranial and intraspinal epithelial cysts: a simplified classification]. PMID- 10844365 TI - [Description and analysis of olfactory disorders after head trauma. Review of the literature]. AB - Olfactory disorders subsequent to head trauma can raise medicolegal problems when the olfactory deficit occurs after a work accident or a traffic accident and also raises the problem of psychological and functional tolerance when the patient consults late for a therapeutic solution. We studied the epidemiology of olfactory disorders after head trauma. Such disorders have been recognized since the XIX(th) century. Several studies have attempted to determine the prevalence of olfactory sequelae after head trauma. We then describe the pathophysiology of these lesions and emphasize that the degree of severity is related with the gravity of the head trauma and that the prevalence of these disorders varies considerably depending on the site of the trauma. Mechanisms which can lead to lesions of the olfactory system include, damage to the olfactory nerve bundles, naso-sinus lesions, or lesions of the cerebral olfactory centers. Brain MRI in anosmic patients after head trauma allows a visualization of intracranial lesions. The degree of reversibility of olfactory disorders after head trauma is imperfectly evaluated and discordant results have been reported. Some authors consider loss of olfaction is definitive. Others suggest an improvement in olfaction the year following head trauma. We reviewed the literature on these different elements. Expert opinions concerning patients complaining of olfactory disorders should relate this disorder to the initial trauma and determine the personal and occupational impact of the deficit. Attribution of cause must take into account all the clinical, radiological and psycho-olfactory factors. PMID- 10844366 TI - [A neuropsychological and functional brain imaging study of visuo-imitative apraxia]. AB - We describe the case of a 58-years-old right-handed women suffering from an occipital-parietal lesion. The administration of a cognitively based assessment tool for limb praxis (Batterie d'Evaluation des Praxies, B.E.P., Peigneux and Van der Linden, 1998) demonstrated bilateral visuo-imitative apraxia. Gesture production was mainly characterised by spatial, errors, and imitation of meaningful gestures was worse than their pantomime on verbal command. Moreover, the imitation of meaningless gestures and their reproduction on a manikin were worse than imitation of their matched meaningful gestures. In a cognitive perspective, adapted from the Rothi et al. (1997) and Goldenberg (1995) contributions to our understanding of limb praxis, this configuration of performance suggests deficits occurring at multiple levels. On one hand, it suggests either access difficulties or alteration of the output praxicon, i.e., the lexicon for visuo-kinesthetic engrams of meaningful gestures. On the other hand, the simultaneous deficit for meaningless gesture reproduction on the subject's own body and on a manikin favors an alteration of the structural descriptions of the human body (i.e., human body knowledge), underlying the mental transposition processes occurring between the visual analysis of a meaningless gestural configuration and its effective reproduction on oneself or on a manikin, thus contradicting the classic view of a direct pathway linking visual analysis and motor planning in meaningless gesture imitation. Finally, due to the output praxicon deficit, imitation of meaningful gestures is partly processed in the same way as meaningless gestures (also impaired in this case), leading to an interference effect between both degraded memory-based and visually transposed traces, which account for imitation of meaningful gestures being worse than their pantomime on verbal command. We also assess regional cerebral metabolism using positron emission tomography (PET). Comparison with 41 healthy subjects (SPM96) demonstrated a statistically significant hypometabolism in the left intraparietal sulcus and superior parietal lobule, and in the right dorsal prestriate cortex. These results, together with a review of the other studies of visuo-imitative apraxia, suggest that the left intraparietal sulcus may be associated with access or integration of information from the output praxicon. The left superior parietal and the right dorsal prestriate deficits functionally impaired a bilateral dorsal network implied in the mental transformations of the body, thus suggesting that these mental transformations are underlined by knowledge of the human body, which may subsequently explain the deficit for the reproduction of meaningless and meaningful configurations. PMID- 10844367 TI - [Epilepsies and time to diagnosis. Descriptive results of the CAROLE survey]. AB - CAROLE is a prospective survey of children and adults who experienced epileptic unprovoked seizure(s) diagnosed for the first time between May 1 1995 and June 30 1996 by 243 French neurologists and neuropediatricians. Case records forms at entry allowed to compare patients who had a single seizure or several seizures prior to diagnosis. In patients with recurrent seizures, the time elapsed between the onset of attacks and the diagnosis (diagnostic delay) was looked for. Half of the 1942 included individuals already experienced more than one seizure when diagnosed. Due to natural history of epilepsies, 13 p.100 of the patients did not come to medical attention after a single seizure. Time to diagnosis ranged from 0 day to 52 years. While the overall median delay was 6 days, it ranged from 0 day to 7-8 months according to the type of seizure and the epilepsy syndrome. Two thirds of generalized convulsive seizures were immediately diagnosed versus one third of partial seizures. One half of infantile spasms were identified within 2 weeks versus 6 weeks in complex partial seizures, 4 months in absence seizures, 6 months in simple partial seizures, and 7 months in myoclonic seizures. Three quarters of idiopathic partial epilepsies were diagnosed within 4 weeks versus 3 months in symptomatic generalized epilepsies, 8 months in symptomatic partial epilepsies, 15 months in idiopathic generalized epilepsies, 30 months in cryptogenic partial epilepsies, and 33 months in undetermined epilepsies. So, the time elapsed between a first epileptic event and its diagnosis is epilepsy dependent: seizure type and cause. Other reasons of diagnostic delay do exist: doctor and patient. They will be addressed in another study. PMID- 10844368 TI - [Spinal cord magnetic resonance imaging in multiple sclerosis: importance of determining degree of atrophy as a marker of disease course]. AB - Spinal cord magnetic resonance imaging (MRI) is of particular interest in the management of multiple sclerosis (MS) especially in primary progressive forms. Most of the demyelinating lesions are located in the cervical or dorsal cord. Spinal cord area reduction has been recently correlated with the progression of disability (Losseff et al., 1996, Lycklama a Nijeholt et al., 1998). The aim of this study was to confirm this first result, to assess the reproducibility of this method and to correlate demyelinating lesions with spinal cord area reduction. Fifty two patients were included and compared with 15 controls (normal subjects). T2 Sagittal and axial plane images were performed to localized hypersignal lesions. Spinal cord area was obtained by a volume acquired inversion prepared fast spoiled gradient echo acquisition (MP-Rage) sequence. We compared the mean area value with clinical parameters (age, course of the disease, expanded disability status scale ?EDSS) and with the number and location of demyelinating lesions. Demyelinating lesions were found in 82p.100 of MS patients and in none of controls. Mean spinal cord area was closely similar to Losseff et al. (1996) results and was reduced compared with controls (p<0.001). Spinal cord reduction was correlated with disability, studied by the EDSS. Furthermore, no correlation was found between demyelinating lesions and spinal cord area reduction. This study confirms the interest of spinal cord area mesurement in MS. Spinal cord atrophy is a reliable marker for axonal loss. This method should be of particular interest for the follow-up of axonal loss in thepeutic trials especially in primary progressive MS. PMID- 10844369 TI - [Familial myopathy with desmin storage seen as a granulo-filamentar, electron dense material with mutation of the alphaB-cristallin gene]. AB - Two familial cases of a myopathy remarkable by the presence of a granulo filamentar, electron dense material were reported in 1978. In a second step, in 1988, it was demonstrated that this material contained an abnormally phosphorylated desmin. During the last twenty years, the occurrence of new cases in this family confirmed the autosomal dominant inheritance of the disease, and made it potentially informative for molecular genetics studies. This allowed first to map the disease on chromosome11q21-23, and afterwards to identify a mutation within a gene coding for a chaperone protein, alphaBcrystallin. An extensive clinical, pathological and genetic study of this princeps family is herein reported in detail. First, it showed the possible detection of histopathological changes in presymptomatic patients. Second, it allowed to demonstrate the simultaneous occurrence of both alphaBcrystallin and desmin in the granulo-filamentar aggregates. Third, this study provided a precise knowledge of the evolution rate of the disease. The analysis of similar observations reported in the literature clearly shows the clinical, pathological and genetic heterogeneity of this new neuro-muscular disorder. PMID- 10844370 TI - [Psychiatric presentation of human African trypanosomiasis: overview of diagnostic pitfalls, interest of difluoromethylornithine treatment and contribution of magnetic resonance imaging]. AB - We report a case of a western African man, residing in France for 4 years, who developed human African trypanosomiasis (HAT) caused by Trypanosoma brucei gambiense. Diagnosis was made at a late stage of the disease. The disease was misdiagnosed and untreated for several years because the clinical presentation was limited to psychiatric disorders and biological confirmation was difficult. Polysomnographic recordings demonstrated typical alterations of HAT. Difluoromethylornithine was effective in this late stage of the disease. Magnetic resonance imaging showed brain edema with demyelination and associated brain atrophy and abnormal signals in the brainstem and thalamus, both implied in sleep wake cycle. PMID- 10844371 TI - [Spinal cord compression secondary to extramedullary hematopoiesis in a patient with thalassemia]. AB - A 19-year-old man with homozygous beta thalassemia presented with signs of thoracic spinal cord compression secondary to extramedullary hematopoiesis. The patient was treated with hypertransfusion and hydroxyurea. After two months, clinical signs had resolved and magnetic resonance imaging showed significant regression of the extradural mass. Pathophysiology and therapeutic options in this condition are briefly discussed. PMID- 10844372 TI - [Ciguatera and peripheral neuropathy: a case report]. AB - Ciguatera is the most frequently observed form of tropical fish poisoning. It appears as a syndrome associating general signs, gastrointestinal, cardiac and neurological problems. Peripheral and central nervous system signs may be observed. We report a case of a 60-year-old man who developed Ciguatera poisoning with diarrhea, facial paresthesia, myalgia, cramps and weakness. Physical examination revealed a motor distal deficit of the four limbs, myokymia and ataxia. EMG testing was in favor of an axonal neuropathy. Neurologic symptoms persisted for two months. This case illustrates a new pathophysiological mechanism of neuropathy: "axonal channelopathy. Abnormalities of peripheral nerve sodium and potassium channels result in clinical and electrophysiological manifestations unrelated to axonal degeneration or demyelinization. The ciguatoxin mainly acts on sodium channels. Prolonged sodium channel activation results in repetitive axon firing. Recently ciguatoxin was recently demonstrated to have a novel action, blocking the sodium channel leading to slowed nerve conduction and decreased motor and sensory action potential amplitudes. PMID- 10844373 TI - [Corticosteroid-induced epidural lipomatosis]. AB - Epidural lipomatosis is a rare disease, most often associated with chronic administration of exogenous steroids. We report the case of a 69-year-old man treated with 20mg prednisolone daily for 11 years because of polyarthritis. The patient complained of back pain associated with severe weakness of his lower limbs and brisk reflexes, with a progressive onset over one year. MRI showed epidural lipomatosis narrowing the epidural spaces from T-3 to T-10. Tapering of steroids was associated with slow but positive resolution of his symptoms. PMID- 10844375 TI - [ [In Process Citation] PMID- 10844376 TI - [ [In Process Citation] PMID- 10844374 TI - [Painless abducens palsy disclosed by spontaneous dissection of the intracavernous internal carotid artery]. AB - A 63-year-old man presented suddenly and spontaneously an isolated painless oculomotor palsy of the nerve abducens. As no etiology could be suspected a head MRI was performed. It showed a T1 hypersignal of the intracavernous segment of the internal carotid artery with a double-lumen pattern typical of dissection. The patient was treated with aspirin and recovered from clinical symptoms in 10 weeks. A control MRI was performed 3 months after the onset of symptoms and showed the regression of the images of dissection. There was no ischemic lesion of the brain. This case underlines the diversity of the symptoms of spontaneous dissections of cervicocephalic arteries, especially absence of pain, palsies of cranial nerve that are not always limited to lower cranial nerve, and existence of dissections limited to the intracranial segment of the carotid artery. It suggests the interest of MRI in the diagnosis of isolated oculomotor nerve palsies. PMID- 10844377 TI - [The Unified Parkinson's Disease Rating Scale]. PMID- 10844379 TI - News - american academy of neurology : reports of congress of san diego PMID- 10844378 TI - [Converging opinions of a multidisciplinary group of French experts on diagnostic and therapeutic modalities for Alzheimer type dementia]. PMID- 10844380 TI - Relationship between endogenous sex hormone levels, lipoproteins and coronary atherosclerosis in men undergoing coronary angiography. AB - This study was carried out in order to investigate the relationship between endogenous sex steroid hormones and coronary artery disease (CAD). Three hundred and thirty-seven men undergoing coronary angiography were enrolled in the study. Total testosterone, estradiol, free testosterone levels in men with CAD (n = 213) were compared to those of men without CAD (n = 124). No significant differences were found in the serum concentrations of estradiol, total and free testosterone and serum lipid profile between the two groups. Total and free testosterone were negatively (p < 0.05 to p < 0.001) and estradiol was positively (p < 0.05) correlated with age in both groups. Total cholesterol and low-density-lipoprotein levels were positively correlated with the level of free testosterone (r = 0.221, p < 0.01; r = 0.173, p < 0.05, respectively), and high-density-lipoprotein levels were negatively correlated with total testosterone in patients with CAD (r = 0.166, p < 0.05). The results of this study do not support the role of sex steroid hormones in CAD. However, the relationship between sex steroids and serum lipids needs further clarification. PMID- 10844381 TI - Effects of angiotensin-converting enzyme gene polymorphism on the left ventricular function and mass in patients with acromegaly. AB - In this study we have investigated the contribution of the ACE genotype to the development of left-ventricular hypertrophy (LVH) and systolic and diastolic dysfunctions in acromegalic patients. The study group consisted of 30 acromegalic patients (21 women and 9 men, age: 37.9 +/- 10.8 years, disease duration: 9.0 +/- 6.9 years). The distribution of the DD, ID and II genotypes was 40.0 (n = 12), 46.6 (n = 14) and 13.3% (n = 4), respectively, being similar to frequencies observed in a healthy population. Plasma ACE levels were 55.0 +/- 12.0 (45-84), 28.7 +/- 15.7 (8-58) and 24.5 +/- 12.0 (16-33) U/I in patients with the DD, ID and II genotype, respectively. The mean serum ACE activity in the DD genotype was significantly higher than in the heterozygous group (p < 0.0001). Serum ACE activity showed a significant negative association with the mean growth hormone level (r = -0.52, p = 0.007). The LV early diastolic flow velocity/LV presystolic flow velocity (E/A) ratios were 1.2 +/- 0.4 for the DD genotype, 1.3 +/- 0.3 for the ID genotype and 0.7 +/- 0.1 for the II genotype. The E/A ratio was considerably lower in acromegalic patients with the II genotype compared to the other genotypes (p = 0.03). The LV mass index (LVMI) values were 131.5 +/- 4.2 g/m2 for the DD genotype, 141.7 +/- 50.3 g/m2 for the ID genotype and 159.6 +/- 48.2 g/m2 for the II genotype. However, there was no significant difference in LVMI among allelic groups. All other indices of systolic and diastolic function were not statistically different in the acromegalic patients. The present data fail to support a role of ACE gene polymorphism in determining LVH in acromegalic patients. However, the I allele may prove as a useful marker predicting the development of diastolic dysfunction in acromegalic patients. PMID- 10844382 TI - Role of coronary angiography in myxoma patients: a 14-year experience in one medical center. AB - Cardiac myxoma is the most common form of primary heart tumor and often treated with surgical resection without a preoperative angiographic examination for fear of potential risk of sudden death. During the last 14 years, 24 of 38 patients with myxoma underwent coronary angiography. Coronary artery disease (CAD) and other abnormalities were found in 5. Our findings indicated that CAD is not uncommon among the myxoma patients, and coronary angiography should be performed preoperatively in all cases. PMID- 10844383 TI - Cardiac effects of persistent hemodialysis arteriovenous access in recipients of renal allograft. AB - In hemodialysis patients, large arteriovenous (AV) fistulas for vascular access may cause ventricular hypertrophy and high-output cardiac failure. The long-term cardiac consequences of functional AV fistulas in renal transplant patients are unclear. A precise knowledge of these consequences is important to decide if and when such fistulas should be closed in successfully transplanted patients. In this retrospective study including 61 stable renal transplant patients with adequate renal function (serum creatinine <2.0 mg/100 ml), echocardiography was performed in 39 patients with a functional AV fistula (group 1) and in 22 whose fistulas had been closed, for esthetic reasons, within 2 months postoperatively (group 2). The volume flow of the fistulas, measured in 22 randomly selected individuals of group 1, was 900 +/- 350 ml/min (range 500-1,600). Patients of group 1 were older (40 +/- 12 vs. 33 +/- 12 years, p < 0.05), had longer duration of the fistula (62 +/- 31 vs. 36 +/- 30 months, p < 0.05), higher body mass index (24 +/- 4 vs. 22 +/- 3 kg/m2, p < 0.05), systolic (154 +/- 24 vs. 138 +/- 18 mm Hg, p < 0.05) and diastolic (96 +/- 12 vs. 89 +/- 11 mm Hg, p < 0.05) blood pressure and increased left ventricular (LV) end-diastolic dimension (53 +/- 5 vs. 49 +/- 5 mm, p < 0.01). LV mass, cardiac index, ejection fraction and the proportion of patients with LV hypertrophy were comparable in the two groups. LV end-diastolic dimension was positively and independently influenced only by the presence of the AV fistula (p < 0.01) after adjusting for age, duration of the fistula, body mass index, systolic and diastolic blood pressure and the nature of the antihypertensive drugs used. In conclusion, the persistence of large, high flow AV fistulas for prolonged periods of time had little impact on cardiac morphology and function of stable renal transplant patients with adequate renal function. The data do not support routine closure of these fistulas in all renal transplant patients. PMID- 10844384 TI - Clinical utility of available methods for determining platelet function. AB - Determination of platelet function has become increasingly important as new antiplatelet drugs are being developed. Objective means to monitor the pharmacological effects of these agents, in order to maximize efficacy and minimize toxicity, is needed. The available methods for determination of platelet function at the current time are reviewed, including platelet aggregometry, the PFA-100, flow cytometry, platelet contractile force, the rapid platelet function assay, the clot signature analyzer, Sonoclot, and thromboelastography. The principle, advantages, disadvantages, and clinical utility of each method is presented. The uses of the various methods of determining platelet function reported in the literature are summarized, in an effort to provide information regarding the applicability of those different methods. PMID- 10844385 TI - Changes in the mRNA levels of delayed rectifier potassium channels in human atrial fibrillation. AB - INTRODUCTION: We measured mRNA levels of delayed rectifier potassium channels in human atrial tissue to investigate the mechanism of the shortening of the atrial effective refractory period and the loss of rate-adaptive shortening of the atrial effective refractory period in human atrial fibrillation. METHODS AND RESULTS: A total of 34 patients undergoing open heart surgery were included. Atrial tissue was obtained from the right atrial free wall, right atrial appendage, left atrial free wall and left atrial appendage, respectively. The mRNA amounts of KVLQT1 (IKs), minK (beta-subunit of IKs), HERG (IKr), and KV1.5 (IKur) were measured by reverse transcription-polymerase chain reaction and normalized to the mRNA amount of GAPDH. We found that the mRNA levels of KV1.5, HERG and KVLQT1 were all significantly decreased in patients with persistent atrial fibrillation for more than 3 months. In contrast, the mRNA level of minK was significantly increased in patients with persistent atrial fibrillation for more than 3 months. We further showed that these changes were independent of the underlying cardiac disease, atrial filling pressure, gender and age. We also found that there was no spatial dispersion of mRNA levels among the four atrial sampling sites. CONCLUSIONS: Because the decrease in potassium currents results in a prolonged action potential, the shortening of the atrial effective refractory period in atrial fibrillation should be attributed to other factors. However, the decrease in IKs might contribute, at least in part, to the loss of rate-adaptive shortening of the atrial refractory period. PMID- 10844386 TI - Early time course of heart rate variability after thrombolytic and delayed interventional therapy for acute myocardial infarction. AB - In 89 of 97 consecutive patients with myocardial infarction (MI) undergoing thrombolysis and delayed early coronary angiography with PTCA, if indicated, heart rate variability (HRV) in time domain was evaluable 40 +/- 11 h after the onset of chest pain using 24-hour ECG recordings. Patients with anterior MI (n = 40) had lower values for HRV and left ventricular ejection fraction (p < 0.05). The mean of all 5-min standard deviations of RR intervals (SDNNi) and the root mean-square difference of successive RR intervals (rMSSD) decreased significantly (p < 0.001 each), whereas the standard deviation of all normal RR intervals and the percentage of absolute differences between successive RR intervals only showed a tendency to lower values 4 weeks after MI (p = 0.20 and 0.08, respectively). The decreases in SDNNi and rMSSD were more evident in inferior than in anterior MI. The time course of HRV following MI was similar in patients with and without PTCA. These results indicate an initial vagal hyperactivity in inferior MI, which is quickly predominated by sympathetic activation and a prolonged recovery of the cardiac autonomic imbalance after MI despite a successful combined reperfusion therapy. PMID- 10844387 TI - Visual versus computerized analysis of upsloping ST segment depression in the exercise electrocardiogram. AB - A slowly upsloping ST segment depression is an abnormal, and a rapidly upsloping ST segment depression is a normal exercise ECG response. We investigated the agreement of expert physicians on the visual classification of the ST segment depression, and compared the (majority) vote with the computer-generated ST slope. A total of 206 exercise ECG leads with an amplitude of the ST segment depression > or = 0.15 mV and a ST segment slope > or = 0.5 mVs(-1) were evaluated. All three interpreters agreed in 68 cases, two agreed in 123 cases, and all disagreed in 15 cases. Intraobserver agreement was 61%. The ST segment slope was significantly (p < 0.001) greater in leads generally interpreted as rapidly upsloping (n = 38; 2.1 +/- 0.8 mVs(-1)), than in those interpreted as slowly upsloping (n 121; 1.3 +/- 0.6 mVs(-1)) or horizontal (n = 32; 1.1 +/- 0.4 mVs(-1)), although there was some overlap. Thus, standardization of the computer assisted exercise ECG interpretation should be continued. PMID- 10844388 TI - Atrial ejection force in systemic autoimmune diseases. AB - Systemic autoimmune disorders may affect several organs, including the heart. We analyzed two-dimensional and pulsed Doppler echocardiograms of patients (n = 37) with systemic lupus erythematosus (SLE, n = 24) or rheumatoid arthritis (RA, n = 13) to determine whether atrial ejection force (AEF) could represent a suitable parameter for detecting left ventricular filling abnormalities in SLE and RA. In both patient subgroups, AEF was significantly higher than in healthy controls (n = 40) matched for gender and age (14.0 +/- 5.4 vs. 11.0 +/- 3.5 kdyn, p < 0.01). Because conventional echocardiographic parameters of left ventricular function failed to detect such a difference, AEF might serve as an additional sensitive parameter for detecting left ventricular diastolic filling abnormalities early in the course of a systemic autoimmune disease. PMID- 10844389 TI - An interleukin-6-producing cardiac myxoma associated with mediastinal lymphadenopathy. AB - We report our experience with a patient whose mediastinal lymphadenopathy resolved after resection of a cardiac myxoma that secreted interleukin-6 (IL-6). The patient was a 68-year-old female who complained of nocturnal chest discomfort related to congestive heart failure. An echocardiogram demonstrated a large left atrial mass. A computed tomogram showed not only the left atrial mass but multiple enlarged mediastinal lymph nodes. The serum IL-6 level was markedly elevated at 13.7 pg/ml. After resection of the cardiac myxoma, serum IL-6 returned to the normal range. A repeat computed tomogram showed no mediastinal lymphadenopathy. We believe that overproduction of IL-6 by the cardiac myxoma was the cause of the mediastinal lymphadenopathy. PMID- 10844390 TI - Cold pressure test producing coronary spasm, coronary thrombosis and myocardial infarction in a patient with IgM antibodies against Coxsackie B virus. AB - Several lines of evidence have shown that viral infections are capable of causing coronary spasm and precipitating or mimicking clinical myocardial infarction. Here we report the case of a 41-year-old woman with recurrent angina who was admitted to our hospital because of ventricular tachycardia. Laboratory examination revealed positive IgM titers against Coxsackie B virus. Coronary angiography showed normal coronary arteries, but following a cold pressure test severe spasm of all coronaries with thrombotic occlusion of the second marginal branch of the circumflex artery occurred. We conclude that coronary spasm should be clinically suspected in patients with chest pain and ventricular arrhythmia in combination with IgM antibodies against Coxsackie B virus. In these patients, a cold pressure test should be avoided, and antithrombotic and antispastic therapy is recommended. PMID- 10844391 TI - The oldest patient to undergo aortic valve replacement. AB - As the American population ages, more elderly patients will present with significant heart disease. This group has an increase in morbidity and mortality with heart surgery as compared to a younger population. Severe aortic stenosis has a grave prognosis, and therefore surgical options should be considered even in the most elderly patient with this condition. We present the oldest reported patient to undergo aortic valve replacement for severe aortic stenosis. PMID- 10844392 TI - Highly effective compensatory mechanisms in a 76-year-old man with a coarctation of the aorta. AB - We present a man who was diagnosed with coarctation of the aorta (CA) at the age of 76. He had developed an extensive collateral circulation for maintaining the circulation below the CA. This compensatory mechanism was highly effective, since the patient had remained asymptomatic and in good health during most of his life. He died from intrahospitalary pneumonia, unrelated to CA. Long-term survival in patients with untreated CA is exceptional, with only 11 patients living into their 70s being previously reported. A review of these cases is included. PMID- 10844393 TI - The use of Xenopus laevis oocytes for the functional characterization of heterologously expressed membrane proteins. AB - The oocytes of the South African clawed frog X. laevis are widely used for the expression of heterologous proteins. The functional characterization of membrane proteins in particular has significantly profited from the use of this expression system. Heterologous cRNA can easily be injected and protein expression and function be studied with several techniques. This review will give a short overview into the variety of methods applicable. They span from different electrophysiological methods such as two electrode voltage clamp, patch clamp and ion-selective electrodes over cytochemistry to protein biochemistry. In spite of the wide usage of Xenopus oocytes, caution should be taken interpreting the results of protein expression. Heterologous proteins may either interact with endogenous proteins, the background of endogenous protein function may be relatively high, or altered protein behaviour may occur due to differences of the ambient temperature or altered cellular environment. PMID- 10844394 TI - Secretin increases the paracellular permeability of CAPAN-1 pancreatic duct cells. AB - The effects of secretin, the physiological secretagogue for pancreatic ducts, were studied in CAPAN-1 pancreatic duct carcinoma cells. When grown to confluence on plastic dishes, CAPAN-1 cells form domes and exhibit marked increases in culture content of Na+ and urea distribution space (UDS). This parameter is measured as an index of both intracellular and dome compartments under the conditions adopted. Both Na increase and dome formation are inhibited by long term incubation with phorbols, DIDS, DPC, EIPA, H2DIDS, and brefeldin. Short term treatment with secretin or 8-Br-cAMP/teophylline causes dome collapse and a marked decrease in UDS and culture content of Na. Secretin-induced sodium decrease is not abolished by ion channel inhibitors, suggesting that diffusion routes other than ion channels are involved in hormone effects. This hypothesis is also in agreement with data obtained on CAPAN-1 cells cultured on permeable inserts, where no change in Na content or UDS is detected upon secretin treatment. Confluent monolayers exhibit a high transepithelial resistance (Rms) which is markedly and reversibly decreased by secretin. The hormone also decreases the transepithelial voltage (Vms) and raises the monolayer permeability to mannitol. It is concluded that secretin enhances the paracellular permeability of pancreatic duct cells. This effect of secretin, unknown thus far, may be involved in the mechanism of pancreatic secretion in vivo. PMID- 10844395 TI - Alpha integrin subunits regulate human (Caco-2) intestinal epithelial proliferation and phenotype. AB - BACKGROUND/AIMS: Cell-matrix interactions influence intestinal epithelial biology, but the whether specific integrin heterodimers exert different effects is unclear. METHODS: We used functional antibodies to investigate effects of the alpha2, alpha3, alpha5, and alpha6 integrin subunits on proliferation and differentiation of human intestinal Caco-2 cells on laminin. Cells seeded onto laminin-coated inserts in defibronectinized medium were treated with functional antibodies or normal IgG for 72 hrs and proliferation, alkaline phosphatase and dipeptidyl dipeptidase specific activity were measured. RESULTS: Caco-2 adhesion to antibody to each alpha-integrin subunit stimulated tyrosine phosphorylation of Focal Adhesion Kinase and paxillin, suggesting that these antibodies intiate integrin-related tyrosine signalling. Proliferation was inhibited by anti-alpha2 and anti-alpha3, but stimulated by anti-alpha6. Alkaline phosphatase and dipeptidyl dipeptidase specific activity were promoted by alpha2 blockade but decreased after a6 blockade. Proliferative blockade using mitomycin C or hydroxyurea prevented the effects of alpha2 ligation on differentiation, but the decrease in alkaline phosphatase specific activity observed after a6 integrin subunit blockade was preserved even after proliferative blockade. CONCLUSION: lntegrin heterodimers modulate human Caco-2 intestinal epithelial biology on laminin in an alpha-subunit specific manner. The different effects of anti-alpha2 and anti-alpha6 may reflect competitition by these antibodies with integrin subunit interactions with laminin as well as initiation of their own signals or different functions for the alpha6 integrin subunit PMID- 10844396 TI - Diadenosine polyphosphates and atrial natriuretic peptide are antiproliferative in rat mesangial cells. AB - Modulation of cell proliferation by vasoactive hormones and growth factors involves also changes in the activity of pH-regulatory transporters. In a preceeding paper (1) we examined the influence of such factors on cellular pH Here the influence of the same factors, diadenosine polyphosphates (ApnA), atrial natriuretic peptide, the growth factor PDGF and the Ca2+-ATPase inhibitor thapsigargin, on proliferation of cultured rat mesangial cells was examined by quantification of [3H]-thymidine incorporation. Mesangial cells were synchronised and growth reduced (0.5% FCS for 24 h) before experiments were started, Incubation with Ap3A, Ap4A, Ap5A or Ap6A (all 10 microM) for 24 h all reduced cell proliferation by 30 to 45%. At 0.1 and 1 microM the effects of Ap4A, Ap5A and Ap6A did not reach significance The antimitogenic effect of Ap5A was not significantly different when cells were incubated for 24, 48 or 72 h. In addition there was no significant difference between the antiproliferative effect of Ap5A in cells of the second, sixth or thirteenth passage. The growth factor PDGF-BB (0.25 nM) resulted man approximately 3-fold increase in [3H]-thymidine incorporation. This increase in proliferation could be significantly reduced by coincubation with 10 microM Ap5A. The mitogenic effect of PDGF was completely abolished in the presence of the Ca2-ATPase inhibitor thapsigargin (1 nM), which also significantly reduced basal cell proliferation by approximately 40%. Incubation of mesangial cells with 10 nM ANP for 24 h reduced basal [3H] thymidine incorporation slightly by approximately 20% and decreased the PDGF induced stimulation. The antimitogenic effects of these agonists is especially pronounced when cells are stimulated. PMID- 10844397 TI - Role of calcium in gentamicin-induced mesangial cell activation. AB - Gentamicin-induced decreases in glomerular filtration rate have been associated to a marked decline in the glomerular capillary ultrafiltration coefficient which could be due to an active contraction of mesangial cells. In the present work we assessed a possible role of cytosolic Ca2+ as a mediator that leads to contraction and proliferation induced by gentamicin on mesangial cells. Gentamicin (10(-5)M) induced an increase in cytosolic free Ca2+, that was fully inhibited by the calcium channel blocker, verapamil, and by the endoplasmic reticulum calcium release blocker, TMB8. Gentamicin induced a planar surface area reduction in cultured mesangial cells, that was blunted by verapamil and TMB-8. Gentamicin also stimulated [3H]thymidine incorporation into DNA and increased viable cell number, effects that were reduced by both, verapamil and TMB-8. Gentamicin stimulated the expression of the AP1 protein; this expression was partially blunted by verapamil and TMB-8. Moreover, verapamil inhibited gentamicin-induced PAF synthesis from mesangial cells. In summary, gentamicin directly raised intracellular Ca2+ activating both calcium influx from external medium and calcium release from internal stores. This increase is responsible of cellular activation (contraction and proliferation) and PAF synthesis induced by gentamicin on mesangial cells. PMID- 10844398 TI - Activation of MAP kinase after reversible ATP depletion in LLC-PK 1 cells. AB - We examined the effect of recovery following reversible ATP depletion on MAP kinase activity in cultured renal cells of proximal tubular origin (LLC-PK1). We induced ATP depletion by 0.1 micromol/l antimycin A in combination with substrate deprivation, and obtained recovery by restoration of substrate supply. MAP kinase activity increased from 374+/-45 pmol/mg protein/mm during ATP depletion to 768 +/- 77 pmol/mg protein/mm after 15 min of recovery. We used ATP to activate a representative G-protein coupled receptor, or epidermal growth factor (EGF) to activate receptors with intrinsic tyrosine kinase activity, and measured the effect of these manipulations on MAP kinase activity during ATP depletion or following recovery. ATP and EGF stimulated MAP kinase activity under control conditions, but not during ATP depletion or after recovery. This shows that two distinct signal transduction pathways represented by ATP and EGF are blocked during ATP depletion and recovery. The lack of energy during ATP depletion and the already maximally stimulated MAP kinase during recovery is likely to be the reason for these results. In summary, these findings suggest that MAP kinase may be involved in the physiological response of cells injured by hypoxia. PMID- 10844399 TI - Effects of adrenaline and tolbutamide on insulin secretion in INS-1 cells under voltage control. AB - The aim of the present study was to investigate whether mechanisms distal to the regulation of Ca2-influx are involved in tolbutamide-induced stimulation and adrenaline- and somatostatin- induced inhibition of insulin secretion in INS-1 cells. Using the patch clamp method, the membrane voltage was either kept constant at -70 mV, or Ca2+-influx was activated by short depolarising pulses to 0 my. These pulses induced an increase in cellular capacitance (Cm) caused by fusion of secretory granules with the plasma membrane. Tolbutamide did not alter, neither Cm under voltage clamp at -70 mV nor increases of Cm due to voltage pulses. The inhibitors of secretion, adrenaline and somatostatin, counteracted the augmentation of [Ca2+]i which was induced by glucose, tolbutamide and forskolin. In the voltage clamp mode, however, where no changes of [Ca2]i. were observed, adrenaline but not somatostatin inhibited the increase of Cm caused by depolarizing voltage pulses. The adrenaline effect on Cm was dependent on the addition of GTP to the pipette solution. When GTP was replaced by GDPbetaS or GTPgammaS, the effect of adrenaline on Cm was abolished. The blockade of calcineurin, by the addition of calcineurin inhibitory peptide (CIP) to the pipette solution, did not affect the adrenaline-induced inhibition of Cm. Moreover. After incubation of the cells with deltamethrin, a calcineurin inhibitor, the stimulation of secretion was attenuated, but the adrenaline induced inhibition was not affected. Our results suggest that adrenaline-induced inhibition of insulin secretion involves a site of action directly related to the exocytotic membrane fusion. In contrast, the stimulator tolbutamide and the inhibitor somatostatin had no direct effect on exocytosis in INS-1 cells. PMID- 10844400 TI - Vimentin in cultured chromaffin cells: an immunofluorescent, biochemical and functional study. AB - In tile present study we seek the presence and possible function of the intermediate filament protein vimentin in adrenomedullary chromaffin cells. Vimentin which is not present in the adrenal medulla was clearly showed up after collagenase digestion of the gland in the cultured chromaffin cells by using an immunofluorescent analysis with double cell labeling with monoclonal antibodies against vimentin and dopamine-beta-hydroxylase. Vimentin was also shown to be phosphorylated in a calcium-dependent manner by acetylcholine. The specific protein phosphatase inhibitor calyculin-A, that has been previously shown to increase vimentin phosphorylation, caused a change in the distribution of vimentin which moved from the Triton X-100 insoluble cytoskeletal preparation to the detergent soluble fraction probably as a result of modifications in filament integrity. The possible role of vimentin in secretion was in addition investigated using digitonin-permeabilized cells, in which the specific antibody for vimentin partially inhibited calcium-induced catecholamine release. These results demonstrate the induction of vimentin expression after collagenase digestion in cultured chromaffin cells and suggest that in these conditions this protein is possibly implicated in the regulation of the secretory process through a phosphorylation-dependent mechanism. PMID- 10844401 TI - Plasma membrane plasticity of Xenopus laevis oocyte imaged with atomic force microscopy. AB - Proteins are known to form functional clusters in plasma membranes. In order to identify individual proteins within clusters we developed a method to visualize by atomic force microscopy (AFM) the cytoplasmic surface of native plasma membrane, excised from Xenopus laevis oocyte and spread on poly-L-lysine coated glass. After removal of the vitelline membrane intact oocytes were brought in contact with coated glass and then rolled off. Inside-out oriented plasma membrane patches left at the glass surface were first identified with the lipid fluorescent marker FM1-43 and then scanned by AFM. Membrane patches exhibiting the typical phospholipid bilayer height of 5 nm showed multiple proteins, protruding from the inner surface of the membrane, with heights of 5 to 20 nm. Modelling plasma membrane proteins as spherical structures embedded in the lipid bilayer and protruding into the cytoplasm allowed an estimation of the respective molecular masses. Proteins ranged from 35 to 2,000 kDa with a peak value of 280 kDa. The most frequently found membrane protein structure (40/microm2) had a total height of 10 nm and an estimated molecular mass of 280 kDa. Membrane proteins were found firmly attached to the poly-L-lysine coated glass surface while the lipid bilayer was found highly mobile. We detected protein structures with distinguishable subunits of still unknown identity. Since X. laevis oocyte is a generally accepted expression system for foreign proteins, this method could turn out to be useful to structurally identify specific proteins in their native environment at the molecular level. PMID- 10844402 TI - Regulation of intracellular pH by bovine intervertebral disc cells. AB - Extracellular acidity is an important determinant of intervertebral disc matrix turnover, possibly exerting effects through changes of intracellular pH (pHi). There is, however, little information concerning the ways in which these cells regulate their pHi. Fluorimetric techniques have been used in the present study to measure pH in isolated intervertebral disc cells, and to characterise the membrane transport pathways by which it is regulated. Nucleus pulposus cells were obtained from bovine intervertebral discs by standard enzymatic digestion techniques, and loaded with the PH-sensitive fluoroprobe BCECF. Resting pHi was approximately 6.7 for cells suspended in either HEPES buffered (HBS) or CO2/HCO3- buffered (BBS) media. Intrinsic buffering capacity was approximately 19 mM pH unit(-1) in HBS and was increased when cells were suspended in BBS. A combination of ion substitution and inhibitor studies for cells at steady-state pH or acidified by exposure to NH4Cl revealed that in HBS Na+ x H+ exchange and an H+ ATPase extrude acid from these cells. Only one of these two systems, the Na+ x H+ exchanger, exhibited a sensitivity to pH, identifying it as the regulator of pH under these conditions. In BBS, an additional pathway which was dependent on extracellular Na+, extracellular HCO3- and intracellular Cl- was detected. These properties are consistent with the four ion HCO3--dependent transporter, although the cation-rich, anion-poor extracellular matrix of the intervertebral disc means that such a pathway has only a marginal role in disc cell pHi regulation. PMID- 10844403 TI - Comparative inhibition of LPS-activated human monocyte-induced thrombin generation by unfractionated heparin and low molecular weight heparins. AB - Monocyte tissue factor may play a role in the physiological or pathological triggering of blood coagulation. It is well known that unfractionated heparin and low molecular weight heparins inhibit extrinsic thrombin generation. However, this notion has never been confirmed in a physiological model using tissue factor obtained from stimulated human monocytes. This is the purpose of this study. It was important to obtain a pure preparation of monocytes with no platelet contamination. This was possible by leukapheresis and elutriation. Under sterile and endotoxin-free conditions, the process does not activate tissue factor expression by monocytes. We adapted the technique of thrombin generation on an automatic analyzer and used human monocyte tissue factor to trigger thrombin generation. Our results show that unfractionated and low molecular weight heparins potently inhibit monocytic tissue factor induced thrombin generation. The comparison of low molecular weight heparins suggests that molecules with higher anti-IIa/anti-Xa ratios exert a stronger inhibitory effect. These data may be relevant to explain the therapeutic effects of unfractionated and low molecular weight heparins in cardiovascular disorders such as unstable angina. PMID- 10844405 TI - Evaluation of the haemostatic system during ketoacidotic deterioration of diabetes mellitus. AB - Clinical observations have indicated the frequent development of thrombotic complications during diabetic ketoacidosis (DKA). This study aimed to examine whether haemostatic changes that could lead to a thrombotic tendency occur during ketoacidosis. Plasma levels of in vivo haemostatic markers reflecting activation degrees of the coagulation system, fibrinolytic system, platelets and endothelium were assayed in 34 patients with DKA, both at diagnosis and 1 week after recovery. We found coagulation system and platelet activation and endothelial injury/activation in the patients at diagnosis of DKA. Although significant improvements were observed after recovery, only platelet activity was completely normalized. Fibrinolytic activity was also increased, both at diagnosis and after recovery, compared to the control group. However, although coagulation activity was prominently increased at diagnosis compared to the recovery period, there was no change in fibrinolytic activity in the same periods; on the contrary, the fibrinolytic capacity of the endothelium was diminished at diagnosis of DKA compared to the recovery period, suggesting the presence of relative hypofibrinolysis during DKA. Indications for a role of hyperglycaemia in the emergence of haemostatic disturbances during DKA were observed. PMID- 10844404 TI - Thromboprophylaxis in hip fracture surgery: a pilot study comparing danaparoid, enoxaparin and dalteparin. The TIFDED Study Group. AB - A pilot study was performed to compare the thromboprophylactic effect of danaparoid, enoxaparin and dalteparin in patients with hip fracture. The study was a prospective, randomised assessor-blind, four-centre trial. Prophylaxis was given for 9-11 days, whereafter bilateral phlebography was performed. A total of 197 patients were randomised. There were no statistically significant differences in the frequency of deep vein thrombosis, in blood loss or bleeding complications between the three prophylaxis groups. In conclusion, this moderately sized study revealed no statistically significant difference in efficacy or safety between danaparoid, enoxaparin and dalteparin in patients undergoing hip fracture surgery. PMID- 10844406 TI - Differential behavior of coagulation factor XIII in patients with inflammatory bowel disease and in patients with giant cell arteritis. AB - The aim of this prospective study was to examine the role of coagulation factor XIII (FXIII) in relation to disease activity in inflammatory bowel disease (IBD) and in giant cell arteritis. Plasma FXIII activity was studied during active and inactive disease in newly diagnosed patients with Crohn's disease (CD; n = 20), ulcerative colitis (UC; n = 18) and giant cell arteritis (GCA; n = 19), in 3 month intervals (median follow-up 12 months). FXIII was also measured in two noninflammatory control groups, age and sex matched for IBD (n = 25) and GCA (n = 26). FXIII activity was significantly lower in active CD or UC than in active GCA or the noninflammatory controls. Both in CD and UC, FXIII activity correlated inversely with indices of clinical disease activity, the erythrocyte sedimentation rate, fibrinogen and C-reactive protein levels. Low FXIII activity is a characteristic feature of active IBD, and serial measurements may be useful to assess IBD activity. PMID- 10844407 TI - Correlation of circulating 17beta-oestradiol with haemostatic factors in healthy postmenopausal women. AB - The aim of the study was to evaluate any correlation between the circulating oestrogenic hormone 17beta-oestradiol and haemostatic factors in healthy postmenopausal women. In keeping with this objective, the correlations were evaluated irrespective of whether the source of the hormone was purely endogenous or exogenous as well. Accordingly, a univariate correlation adjusted for age, body mass index, and duration of menopause was determined in 42 healthy postmenopausal women aged 47-78 years, 19 of whom were self-reported users of hormone replacement therapy. The rest were self- reported never users. Serum 17beta-oestradiol exhibited a direct correlation with endogenous thrombin potential extrinsic pathway (R = 0.42, p = 0.01) and prothrombin fragments 1 and 2 (R = 0.37, p = 0.03) and an inverse correlation with antithrombin III (R = 0.36, p = 0.03) and alpha(2)-antiplasmin (R = -0.45, p = 0.005). The observations suggest an association of this hormone with net thrombin generation on the one hand and improved fibrinolysis on the other. PMID- 10844408 TI - Recombinant thrombomodulin inhibits thrombin-induced vascular endothelial growth factor production in neuronal cells. AB - Thrombin is a serine protease which is generated from its precursor prothrombin by the activation of the blood coagulation cascade. Thrombin converts fibrinogen to fibrin, activates platelets and several coagulation factors, and plays a central role in thrombosis and hemostasis by regulating platelet aggregation and blood coagulation. Here, we show that thrombn enhanced vascular endothelial growth factor (VEGF) production in a dose- and time-dependent manner in the supernatant of cultured PC-12 cells, as determined by enzyme-linked immunosorbent assay (ELISA). Thrombin receptor agonist peptide (SFLLRNPNDKYEPF, TRAP) exerted an effect similar to thrombin on VEGF production. Thrombin-induced VEGF production was significantly attenuated by recombinant human thrombomodulin (rTM) and its minimal functional domain E456. Furthermore, the antioxidant N-acetyl-L cysteine (NAC) markedly inhibited thrombin-induced VEGF production. Thus, rTM and NAC apparently inhibited the effect of thrombin on VEGF production in neuronal cells. PMID- 10844409 TI - Factors influencing the length of a blood trail. PMID- 10844410 TI - Alternative splicing of exon 17 and a missense mutation in exon 20 of the insulin receptor gene in two brothers with a novel syndrome of insulin resistance (congenital fiber-type disproportion myopathy). AB - The insulin receptor (IR) in two brothers with a rare syndrome of congenital muscle fiber type disproportion myopathy (CFTDM) associated with diabetes and severe insulin resistance was studied. By direct sequencing of Epstein-Barr virus transformed lymphocytes both patients were found to be compound heterozygotes for mutations in the IR gene. The maternal allele was alternatively spliced in exon 17 due to a point mutation in the -1 donor splice site of the exon. The abnormal skipping of exon 17 shifts the amino acid reading frame and leads to a truncated IR, missing the entire tyrosine kinase domain. In the correct spliced variant, the point mutation is silent and results in a normally translated IR. The paternal allele carries a missense mutation in the tyrosine kinase domain. All three cDNA variants were present in the lymphocytes of the patients. Purified IR from 293 cells overexpressing either of the two mutated receptors lacked basal or stimulated IR beta-subunit autophosphorylation. A third brother who inherited both normal alleles has an normal muscle phenotype and insulin sensitivity, suggesting a direct linkage of these IR mutations with the CFTDM phenotype. PMID- 10844411 TI - Effects of gender, body composition and birth size on IGF-I in 7- and 8-year-old children. AB - We investigated the relationship between IGF-I, gender, height, weight, body composition and birth size in 260 healthy 7- and 8-year-old children (139 females). All children were born term at Nepean Hospital, Western Sydney. Body composition was measured using dual energy X-ray absorptiometry. IGF-I levels were determined by radioimmunoassay. Girls had higher IGF-I levels than boys (20.2 +/- 6.5 nmol/l compared to 15.9 +/- 6.1 nmol/l, p < 0.001) but there was no correlation between age and IGF-I. IGF-I was positively correlated with height SDS (R(2) = 0.12), weight SDS (R(2) = 0.19), BMI SDS (R(2) = 0.18), total body fat (%) (R(2) = 0.14), and fat-free tissue/cm (R(2) = 0.03). After adjusting for gender and current weight, IGF-I-levels were inversely related to birth size - children with the lowest birth size and heaviest current weight had the highest IGF-I levels. This correlation between birth weight and IGF-I supports the hypothesis that the IGF-I axis is altered in babies who are small for gestational age. PMID- 10844412 TI - Insulin sensitivity is not affected by mutation of codon 972 of the human IRS-1 gene. AB - We investigated the relationship of codon 972 polymorphism of the insulin receptor substrate-1 (IRS-1) gene with insulin resistance in the Japanese population. Among 130 patients with type-2 diabetes mellitus (DM), we identified 6 who were heterozygous for the Gly972Arg mutation. Among 144 healthy subjects, 6 were heterozygous and 1 was homozygous for the mutation. A hyperinsulinemic euglycemic clamp study was performed in 3 of 6 diabetic patients with the heterozygous Gly972Arg mutation and in 60 without it. Both groups showed almost the same levels of insulin sensitivity (glucose infusion rate, GIR = 50.2 +/- 3.0 vs. 51.3 +/- 12.1 micromol/kg/min). Similarly, there was no difference in insulin sensitivity between healthy subjects with and without the mutation using the homeostasis model assessment (HOMA index = 1.14 +/- 0.50 vs. 1.02 +/- 0.63). The frequency of the Gly972Arg allele was not increased in diabetic patients compared with control subjects even in aged (>50 years old) or obese (BMI >/=25) subjects. Among healthy subjects, we identified a 25-year-old male with the homozygous Gly972Arg allele. He was slightly obese (BMI = 25.5) but showed relatively high insulin sensitivity, almost equal to that of healthy subjects without the mutation (GIR = 67.2 vs. 71.8 +/- 22.0 micromol/kg/min). Because the GIR in healthy subjects was significantly higher compared with that in type-2 DM patients, we speculate that another genetic or environmental factor producing a more deleterious effect on insulin sensitivity may exist in diabetic patients. We conclude that this gene abnormality does not play a role in the pathogenesis of insulin resistance and type-2 DM. PMID- 10844414 TI - Lipoprotein(a) levels in formerly small-for-gestational-age children. AB - Lipoprotein(a) (Lp(a)) is an independent and inherited risk factor for coronary artery disease. Concentrations of Lp(a) have been widely described in adolescents, but little is known about its concentration in children born small for gestational age (SGA). To assess the influence of intrauterine growth on Lp(a) levels we examined 50 children born SGA and 21 children born adequate for gestational age (AGA). Lp(a) blood levels (mean +/- SD) of the SGA children differed significantly (p < 0.05) from AGA children (22.3 +/- 22.1 vs. 10.9 +/- 7.6 mg/dl). 14 out of 50 adolescents of the SGA group but 1 out of 21 of the AGA group had elevated Lp(a) (>30 mg/dl) concentrations (p < 0.05). These children also had higher triglyceride (1.0 +/- 0.6 mmol/l vs. 0.74 +/- 0.38 mmol/l) levels (p < 0.05) compared to children with Lp(a) levels <30 mg/dl. Adolescents with Lp(a) levels >30 mg/dl showed a significant inverse relation between Lp(a) levels and gestational age (r = -0.68, p < 0. 005). We hypothesize that impairment of fetal growth might influence serum Lp(a) levels in later life. PMID- 10844413 TI - Activating mutation of GS alpha in McCune-Albright syndrome causes skin pigmentation by tyrosinase gene activation on affected melanocytes. AB - McCune-Albright syndrome (MAS) is a sporadic disease characterized by cafe-au lait spots, polyostotic fibrous dysplasia and hyperfunctional endocrinopathies. To elucidate the mechanism of skin pigmentation, melanocytes, keratinocytes and fibroblasts were primary cultured from the cafe-au-lait spot of a MAS patient. Then, mutational analysis and morphologic evaluation were performed. Also, cAMP level and tyrosinase gene expression in cultured cells were determined. Only Gsalpha mutation was found in affected melanocytes and the cAMP level in affected melanocytes was higher than that of normal melanocytes. The mRNA expression of tyrosinase gene was increased in the affected melanocytes. This study suggests that skin pigmentation of MAS results from activating mutation of Gsalpha in melanocytes and the mechanism involves the c-AMP-mediated tyrosinase gene activation. PMID- 10844415 TI - Medium-term cardiovascular effects of high-dose growth hormone treatment in growth hormone-deficient children. AB - AIM: To investigate the possible cardiac morphofunctional alterations inducd by prolonged and high-dose GH therapy in a group of 14 children with isolated GH deficiency. PATIENTS AND METHODS: Patients were evaluated at phase 1, after 1.1 +/- 0.6 years of treatment with GH 0.93 +/- 0.13 U/kg/week, and at phase 2, after 5.5 +/- 2.1 years of therapy 0.89 +/- 0.11 U/kg/week. At each phase left ventricular volume, mass and systolic function were evaluated by two-dimensional guided M-mode echocardiography; left ventricular diastolic function was assessed by PW-Doppler sampling of transmitral flow. RESULTS: Phase 1: diastolic blood pressure was lower (p < 0.05) and fractional shortening was not adequate for the level of afterload (stress shortening index p < 0.05) in patients compared to controls. Phase 2: diastolic blood pressure was lower (p < 0.01) and mass and mass/volume ratio were increased (mass index p < 0.05, mass/ volume ratio p < 0.05) in patients compared to controls. The increased mass/volume ratio, together with the normal systolic blood pressure, explains the reduction in peak systolic stress (p < 0.005). Among the parameters of left ventricular diastolic function, the peak E velocity/total area under mitral valve tracing and the area under E velocity/total area under mitral value tracing ratios were significantly decreased (p < 0.05). CONCLUSION: After a mean period of 5 years on high-dose GH treatment in GH-deficient children, subclinical morphofunctional alterations in the left ventricle were found. PMID- 10844416 TI - Liquorice, growth retardation and Addison's disease. AB - An 11-year-old boy had hypoparathyroidism and Addison's disease. During treatment with calcitriol, calcium, hydrocortisone and 9-alpha-fluorocortisol, he developed an apparent mineralocorticoid excess and growth retardation. Pseudohyperaldosteronism even persisted after treatment with 9-alpha fluorocortisol was stopped and hydrocortisone was reduced to 6 mg/m(2). The boy reported an excessive daily intake of 300-400 g liquorice corresponding to 600 800 mg glycyrrhizic acid because of salt craving. After complete withdrawal of liquorice all symptoms of hypermineralocorticoidism diminished and growth velocity increased. We hypothesise that inhibition of 11beta-hydroxysteroid dehydrogenase by liquorice caused hypermineralocorticoidism and growth retardation via increased levels of free cortisol in this patient. We conclude that self-medication with liquorice in children with Addison's disease should be considered during treatment. PMID- 10844417 TI - Sonography and scintigraphy are necessary in diagnostics of cystic thyroid lesions. AB - Thyroid sonography in an otherwise asymptomatic 50-year-old woman revealed a mainly cystic lesion within the right thyroid lobe. Thyroid (99)Tc scan showed an enhanced uptake over the right thyroid lobe with suppressed activity over the remaining thyroid tissue. Following aspiration of 6 ml of cyst fluid, a solid lesion became apparent in the location of the original cyst corresponding to the scintigraphically detected autonomous adenoma. This case report emphasizes that ultrasonography and scintigraphy are complementary methods and should be used as such in the primary evaluation of thyroid nodules. PMID- 10844487 TI - Botulinum A exotoxin in cosmetic dermatology. AB - Botulinum A exotoxin, a neurotoxin produced by the bacterium Clostridium botulinum, is now being used by cosmetically oriented specialists for treatment of a large variety of movement associated wrinkles on the face and neck. This form of temporary chemical denervation compliments the cosmetic practitioner's armamentarium alongside resurfacing and tissue augmentation. Additionally, the use of Botulinum toxin to block sympathetic innervation of eccrine sweat glands is proving a valuable treatment of hyperhidrosis of the axillae, palms and soles. PMID- 10844488 TI - The management of oral lichen planus. AB - Oral lichen planus is a relatively common inflammatory disease affecting between 0.5% and 2.2% of the population in epidemiological studies. In contrast with cutaneous lichen planus (LP), in which the clinical course is often mild and resolves within 2 years, mucosal LP tends to follow a more chronic course often punctuated by acute exacerbations. Furthermore, although distinct clinical subtypes such as reticular, atrophic, hypertrophic and erosive forms are well recognized, more than one clinical phenotype may be seen at a time. The rare association with oral neoplasia should always be considered and high-risk patients must be kept under close observation. Thus the management of this disorder will vary widely both between patients, and for individual patients, with fluctuations in disease activity. Here we discuss the therapeutic options available and review the evidence for their use. PMID- 10844489 TI - Treatment of viral warts with cimetidine: an open-label study. AB - The immunomodulatory actions of cimetidine, an H2-receptor antagonist, and its use in the treatment of viral warts has been described previously but its effectiveness is still debated. We report the results in 47 patients with multiple, nongenital viral warts who were treated with oral cimetidine in a 3 month open-label study. The drug was generally well tolerated and 87% of children and 68% of adults improved with treatment. Follow-up data in 65% of the patients showed that there had been no recurrence in the majority of those whose warts had cleared completely during treatment, whereas warts tended to persist or recur in those who had stopped treatment before all the warts had resolved. Our data suggests that cimetidine may be helpful in the treatment of viral warts in both adults and children and supports the need for a randomized controlled trial. PMID- 10844490 TI - Famotidine in the treatment of acute urticaria. AB - Recent studies suggest that histamine H2-receptor antagonists may be useful in the treatment of urticaria. This study was conducted to determine whether famotidine, a H2 antagonist, is effective in the treatment of acute urticaria and compare its effect with that of the H1 antagonist diphenhydramine. In this prospective, double-blind, controlled trial, 25 patients with urticaria of less than 72 h duration were randomized to receive a single dose of either famotidine 20 mg i.m. or diphenhydramine 50 mg i.m. Prior to treatment and 30 min after treatment, patients rated pruritus and sedation using visual analogue scales, while physicians evaluated intensity of urticaria and percentage of body surface area involved by urticaria. Famotidine was found to reduce pruritus associated with acute urticaria, intensity of urticaria, and body surface area affected by urticaria without causing sedation. Famotidine was comparable to diphenhydramine in efficacy; however, there was a (nonsignificant) trend for diphenhydramine to be more effective than famotidine in the treatment of pruritus, and for famotidine to be more effective in the reduction of surface area of involvement. It is concluded that famotidine merits further investigation as a potential medication for treatment of urticaria. PMID- 10844491 TI - The dermatosis of chronic granulomatous disease. AB - A family with X-linked cytochrome-negative chronic granulomatous disease (CGD) involving three generations is reported. The diagnosis of CGD in both the latest male patient and the index male was confirmed by marked impairment in polymorphonuclear leucocyte oxidative burst activity in association with absence of both subunits of cytochrome b. The two female carriers have suffered from chronic inflammatory skin disorders characterized by slowly fluctuating erythematous plaques. The reported cases are discussed in the context of a literature review of the dermatosis of CGD. PMID- 10844492 TI - Successful treatment of severe psoriasis with basiliximab, an interleukin-2 receptor monoclonal antibody. AB - T-cell mediated inflammation plays an important role in the aetiology of psoriasis. We describe a case of severe recalcitrant psoriasis responding well to combined cyclosporin and basiliximab (Simulect(R) Novartis Pharmaceuticals UK Ltd), an interleukin-2 receptor (IL-2R; CD25) chimeric monoclonal antibody. PMID- 10844493 TI - Superior vena cava syndrome presenting as persistent erythematous oedema of the face. AB - The superior vena cava syndrome occurs when extrinsic compression or intraluminal occlusion impedes blood flow through this vessel. Detecting the characteristic cutaneous features can lead to an early diagnosis of this condition. We report a 73-year-old patient with a 4-year-history of erythematous oedema of the face. The underlying cause was a large retrosternal goitre obstructing the superior vena cava. Subtotal thyroidectomy led to dramatic improvement of all dermatologic symptoms. PMID- 10844494 TI - Solitary cutaneous metastasis on the buttock: a disclosing sign of pancreatic adenocarcinoma. AB - A 48-year-old man, previously healthy and asymptomatic, showed a unique skin lesion located on the right buttock. The histopathological study of the lesion disclosed an adenocarcinoma metastatic to the skin. The primary tumour was found at the head of the pancreas, and was confirmed by biopsy. Six months after the beginning of treatment with Gemcitabine the patient is still asymptomatic, and the cutaneous lesion has disappeared. Carcinomas of the pancreas represent less than 5% of human malignant neoplasms, skin involvement is rare, and metastasis generally multiple and situated in the periumbilical area. To our knowledge, very few patients have been reported with a solitary cutaneous lesion disclosing a pancreatic adenocarcinoma. The atypical location, the absence of related symptoms or analytical disorders, and the good progress of the patient make this case especially interesting. PMID- 10844495 TI - Dermatitis herpetiformis effectively treated with heparin, tetracycline and nicotinamide. AB - We report a patient with severe dermatitis herpetiformis (DH) who was intolerant of dapsone, sulphapyridine, systemic steroids, and azathioprine. He was treated effectively with a combination of heparin, tetracycline and nicotinamide. PMID- 10844496 TI - Pseudovascular squamous cell carcinoma of the skin. AB - The presence of acantholysis in squamous cell carcinomas (SCC) may rarely be so extreme that, histologically, it mimics a vascular tumour. However, careful histological examination and immunohistochemical study usually lead to the correct diagnosis. We describe such a case to highlight the clinico-pathological features of this rare form of cutaneous malignancy and to emphasize the difficulties in establishing the correct diagnosis. We also review similar cases reported in the literature. Pseudovascular SCC shows a higher degree of recurrence and metastasis than other variants of SCC. Acantholytic foci in these tumours may demonstrate changes in keratinocyte differentiation markers, and this may explain the more aggresive biological behaviour in the pseudovascular variant of SCC. PMID- 10844497 TI - Treatment of pruritus by capsaicin in a patient with pityriasis rubra pilaris receiving RE-PUVA therapy. AB - Pityriasis rubra pilaris (PRP) is characterized by redness of the skin, scaling and a variable degree of pruritus. We present a patient with extremely itchy PRP successfully treated with oral retinoids and photochemotherapy with 8 methoxypsoralene (RE-PUVA) and topical capsaicin. The PRP-related pruritus which clearly preceded photochemotherapy and for which no other cause was apparent was relieved with capsaicin. This single case report provides evidence that topical capsaicin may be a useful therapeutic option in treating PRP-associated pruritus where antihistamines have been unsuccessful. PMID- 10844498 TI - Generalized atrophic benign epidermolysis bullosa--poor prognosis associated with chronic renal failure. AB - Generalized atrophic benign epidermolysis bullosa (GABEB) is an autosomal recessive form of junctional epidermolysis bullosa, with milder clinical features than the Herlitz subtype. A 25-year-old man presented with the clinical and histological findings of GABEB. At the initial visit, laboratory tests revealed that he also had chronic renal failure (CRF). Usually, GABEB has a good prognosis. However, in this case, the patient had CRF as an associated complication. He died of an intracranial haemorrhage combined with sepsis after 3 weeks of hospitalization. This case suggests that renal complications can occur in this relatively mild form of epidermolysis bullosa, and may contribute to morbidity and premature mortality. PMID- 10844499 TI - Coma-induced bullae and sweat gland necrosis following clobazam. AB - Coma-induced bullae and sweat gland necrosis is a rare clinicopathological entity described in association with a variety of aetiopathological conditions all of which have resulted in an impairment of conscious level. We report the first case observed in association with clobazam, used as adjunctive therapy for resistant epilepsy in a 4-year old. The potential underlying mechanisms and previously reported associations are discussed. PMID- 10844500 TI - Paraneoplastic pemphigus associated with Castleman's tumour. AB - We present a case of paraneoplastic pemphigus associated with Castleman's disease. Our patient had stomatitis and vulvar erosions followed by a cutaneous polymorphous eruption. Investigations for neoplasia disclosed a Castleman's tumour suggesting the diagnosis of paraneoplastic pemphigus. The diagnosis was supported by immunoblotting using an extract of cow tongue although keratinocytes extracts did not identify relevant target antigens. One year after surgical excision of the tumour the patient remained unwell with persistent buccal erosions and lymphopenia. This case is unusual because of the length of its evolution before the discovery of the Castleman's tumour. It is only the second occasion that the association between paraneoplastic pemphigus and Castleman's tumour has been reported. Our case emphasizes the usefulness of immunoblotting on cow tongue in suspected cases of paraneoplastic pemphigus. PMID- 10844501 TI - Tuberculous cellulitis. AB - We report a case of cutaneous tuberculosis presenting as cellulitis. The patient was a 63-year-old Korean woman who also had diabetes mellitus and a 20-year history of oral corticosteroid medication prescribed for arthralgia. In addition, she had had pulmonary tuberculosis 20-year previously for which she received systemic treatment for 1 year. Her clinical cellulitis failed to respond to antibiotic therapy. Subsequent investigations, using histopathology and polymerase chain reaction, established an alternative diagnosis of cutaneous tuberculosis. The skin eruption cleared after treatment with isoniazid, rifampicin, ethambutol and pyrazinamide. This case represents a most unusual presentation of tuberculosis in the skin. The atypical features may reflect the patient's general medical state. PMID- 10844502 TI - An unusual case of pemphigus vulgaris presenting as bilateral foot ulcers. AB - We describe an unusual presentation of pemphigus vulgaris, an autoimmune intraepidermal blistering skin disease associated with autoantibodies to the desmosome glycoprotein, desmoglein 3. A 60-year-old man presented with bilateral ulceration on the dorsum of the feet. These clinical features persisted for 4 months before more characteristic signs of pemphigus vulgaris, including mouth ulceration and skin erosions, developed. The atypical presentation led to a delay in diagnosis and initiation of the appropriate treatment. Pemphigus vulgaris may have unusual manifestations, such as nail dystrophy, paronychia, or granulation tissue-like lesions, but this case of bilateral foot ulceration highlights a further, perhaps unique, clinical presentation of this autoimmune disease. PMID- 10844503 TI - Sporotrichoid presentations in leprosy. AB - Two adult patients of leprosy, one woman and one man, presented with a clinical picture simulating sporotrichosis. The skin and regional nerve trunk was affected in one, and in the other the disease was confined to the nerve. Both had features of an upgrading reaction following anti-leprosy therapy; this was seen as erosion and scarring of the plaque, and acute onset of abscesses along the easily palpable and thickened nerve that ruptured through the skin. The diagnosis was supported by histopathology. In the light of other infections that give rise to a sporotrichoid pattern of infection it is concluded that leprosy should also be included in this category so that early diagnosis and use of corticosteroids can be implemented quickly to prevent nerve destruction. PMID- 10844504 TI - Tumour suppressor genes. PMID- 10844505 TI - Diagnosis of pemphigus by ELISA: a critical evaluation of two ELISAs for the detection of antibodies to the major pemphigus antigens, desmoglein 1 and 3. AB - Pemphigus vulgaris (PV) and pemphigus foliaceus (PF) are characterized by autoantibodies to the desmosomal glycoproteins desmoglein 3 (Dsg 3) and Dsg 1 (Dsg 1), respectively. In this study, two enzyme-linked immunosorbent assays (ELISA) which detect IgG autoantibodies to Dsg 1 and Dsg 3 have been evaluated. A total of 317 normal and disease controls, 82 patients with PV and 25 with PF were studied. The Dsg 3 ELISA was positive in all 34 patients with untreated PV and the Dsg 1 ELISA was positive in all 10 with untreated PF. When patients undergoing treatment were included, the sensitivities fell to 95% and 92%, respectively, but still compared favourably to the sensitivity of indirect immunofluorescence which was 79% in PV and 84% in PF. All PF sera were negative in the Dsg 3 ELISA and the specificity of both assays was 98% or greater. Large numbers of samples could be analysed simultaneously over a relatively short time period. The Dsg 1 and Dsg 3 ELISAs also provided objective, quantitative, reproducible data which allowed differentiation of PV from PF and in view of these advantages, they are likely to become a routine technique in diagnostic laboratories. PMID- 10844506 TI - Epidermolytic hyperkeratosis in a Hispanic family resulting from a mutation in the keratin 1 gene. AB - Epidermolytic hyperkeratosis (EHK; bullous congenital ichthyosiform erythroderma) is a genodermatosis resulting from mutations in either the keratin 1 (K1) or keratin 10 (K10) genes. It is characterized by erythroderma and blistering at birth, and the development of ichthyotic hyperkeratosis and palmoplantar keratoderma. A wide variety of mutations within the highly conserved helix initiation and termination motifs of the central rod domains of the K1 or K10 genes correlate with the highly variable phenotypic severity observed in EHK. We report a novel missense mutation designated L214P in a large Hispanic pedigree with EHK. The mutation is located in the highly conserved 1A segment of the alpha helical rod domain. The presence of this mutation underscores the importance of sequence alterations located in the central rod domain in the pathogenesis of EHK. PMID- 10844507 TI - Epidermolytic palmoplantar keratoderma in a Hispanic kindred resulting from a mutation in the keratin 9 gene. AB - Epidermolytic palmoplantar keratoderma (EPPK) is a localized keratinization disorder caused by mutations in the highly conserved coil 1A domain of the keratin 9 gene, KRT9. We present a Hispanic pedigree spanning three generations, with affected individuals in all generations. Using polymerase chain reaction amplification and direct sequencing we demonstrated a previously reported missense mutation in KRT9, which is expressed almost exclusively in the skin of palms and soles. The C-->T missense mutation R162W changes a basic amino acid (arginine) to a neutral amino acid (tryptophan). We describe this mutation in a Hispanic pedigree with EPPK for the first time, extending the finding of this mutation in other genetic backgrounds, and demonstrating the prevalence of this mutation in diverse populations. PMID- 10844508 TI - Biomechanical measurement of skin distensibility in scleredema of Buschke associated with multiple myeloma. AB - We report a case of scleredema of Buschke associated with IgG kappa monoclonal hypergammaglobulinaemia. After myeloma polychemotherapy an improvement in skin involvement was observed and confirmed by means of noninvasive skin elasticity measurements. This suggests a relationship between the two diseases. The bioengineering method used can be useful for early detection and monitoring the skin involvement in patients with this disease association. PMID- 10844509 TI - Topical tacrolimus in dermatology. AB - Tacrolimus and cyclosporin A are potent immunosuppressants that are used systemically to treat several inflammatory skin conditions successfully. They differ in their structure and tacrolimus is 10-100 times more potent than cyclosporin A. They have similar side-effects. They have been used topically in various clinical studies. Topical cyclosporin A is largely ineffective whereas topical tacrolimus is effective in treating atopic dermatitis. Topical tacrolimus has not been studied sufficiently in treating psoriasis although it has been used successfully in allergic contact dermatitis, erosive mucosal lichen planus and pyoderma gangrenosum. PMID- 10844510 TI - Dermatomyositis-like eruption and leg ulceration caused by hydroxyurea in a patient with psoriasis. PMID- 10844511 TI - Authors' reply PMID- 10844512 TI - Nail dystrophy in association with polydactyly and benign familial hypercalcaemia. PMID- 10844513 TI - Sarcoid panniculitis. PMID- 10844514 TI - Migrating mast cells into the epidermis of wet and inflammatory granuloma. PMID- 10844516 TI - C1q receptors. PMID- 10844517 TI - Disordered expression of inhibitory receptors on the NK1-type natural killer (NK) leukaemic cells from patients with hypersensitivity to mosquito bites. AB - Recent studies have revealed the existence of a distinct type of NK cell leukaemia of the juvenile type, which presents with hypersensitivity to mosquito bites (HMB) as an essential clinical manifestation and is infected with clonal Epstein-Barr virus (EBV). This disorder is thus called HMB-EBV-NK disease and has been reported in Orientals, mostly from Japan. We investigated the profile of cytokine production and the expression of both types of NK inhibitory receptors, i.e. CD94 lectin-like dimers and killer-cell immunoglobulin-like receptors, in NK leukaemic cells from three patients with HMB-EBV-NK disease. It was found that freshly isolated NK leukaemic cells expressed mRNA for interferon-gamma (IFN gamma) and additionally produced IL-10 upon stimulation with IL-2, indicating that the NK cells were of NK1 type. More than 98% of NK cells from the patients bore CD94 at a higher level than did normal NK cells, whereas p70 or NKAT2, belonging to immunoglobulin-like receptor, was not expressed in those NK cells. Freshly isolated leukaemic NK cells transcribed mRNA for CD94-associated molecule NKG2C at an abnormally high level, and upon stimulation with IL-2 and/or IL-12 they expressed NKG2A as well. The disordered expression of these inhibitory receptors not only provides some insights into the pathogenesis of HMB-EBV-NK disease but also can be used as phenotypic markers for the diagnosis of this type of NK cell leukaemia. PMID- 10844515 TI - X-linked immunodeficiency with hyper-IgM (XHIM). PMID- 10844518 TI - Inhibition of aromatic L-amino acid decarboxylase activity by human autoantibodies. AB - A full-length rat cDNA clone encoding aromatic L-amino acid decarboxylase (AADC) (E.C. 4.1.1.28) was used for in vitro transcription and translation. The enzyme had catalytic activity (0. 2 pmol serotonin/microl lysate per min), and was stimulated 2.5-fold by the addition of excess pyridoxal phosphate. On size exclusion chromatography, AADC eluted as a single activity peak with an apparent mol. wt of 93 kD. This activity peak was immunoprecipitated by sera from patients with autoimmune polyendocrine syndrome type I (APS I) containing autoantibodies against AADC. Serum and purified IgG from these patients inhibited the enzyme activity (non-competitively) by 10-80%, while sera from APS I patients without autoantibodies and controls did not. This finding confirms and extends previous observations that APS I patients have inhibitory antibodies against key enzymes involved in neurotransmitter biosynthesis. PMID- 10844519 TI - Neuroendocrine changes in colon of mice with a disrupted IL-2 gene. AB - Neuroendocrine peptides have a variety of physiological functions in the gastrointestinal tract. This study was carried out to investigate the impact of IL-2 deficiency on the neuroendocrine system in normal colon, and the neuroendocrine changes during colonic inflammation. Mice with homozygous disrupted IL-2 gene (IL-2-/-) spontaneously developed a bowel disease with similarities to human ulcerative colitis. Different types of colonic endocrine cells and myenteric nerves were analysed in the IL-2-/- mice using immunomorphometry. The neuropeptide contents in the colonic tissues were determined by radioimmunoassay. Age-matched healthy IL-2+/- and IL-2+/+ mice served as controls and the colonic IL-2 levels were compared between these two groups of mice by ELISA. Our data showed that less than half the amount of IL-2 was synthesized in the colon of IL-2+/- mice compared with the IL-2+/+ wild-type mice. Two major differences in the neuroendocrine colon were found between the mice with an intact and disrupted IL-2 gene. One was age-related. The frequencies of various endocrine cells and myenteric nerves increased with age in the IL-2+/+ mice. However, no such increases were seen in the mice with a disrupted IL-2 gene. Instead, the volume densities of enteroglucagon, serotonin cells and substance P (SP), vasoactive intestinal polypeptide (VIP) and total myenteric nerves were lower in the older IL-2+/- and IL-2-/- mice compared with the wild type. The other was disease-related. Polypeptide YY (PYY) cells and tissue levels of PYY, SP and VIP were significantly decreased in the IL-2-/- mice during the course of bowel inflammation compared with the healthy IL-2+/- and IL-2+/+ controls. These findings indicate that colonic neuroendocrine alterations did occur in the mice with a disrupted IL-2 gene and diminished local IL-2 level, suggesting a role of IL-2 in the regulation of the neuroendocrine system and a prevalent interaction between the immune and neuroendocrine systems in normal colon. On the other hand, there were some changes that seemed to correlate with the bowel inflammatory process. They might be associated with the impaired function in inflamed gut and contribute to the development and/or prolongation of disease. PMID- 10844521 TI - Granulocyte-macrophage colony-stimulating factor (GM-CSF) has opposing effects on the capacity of monocytes versus monocyte-derived dendritic cells to stimulate the antigen-specific proliferation of a human T cell clone. AB - GM-CSF is widely used in combination with IL-4 to differentiate monocytes into potent T cell stimulatory cells, referred to as monocyte-derived dendritic cells (MoDC). These cytokines further increased the stimulatory function of MoDC when present during their incubation with antigen, as determined by the proliferative response of an allergen-specific T cell clone. Conversely, the incubation of freshly isolated monocytes with antigen in the presence of GM-CSF or GM-CSF and IL-4 strongly inhibited the specific stimulation of the T cells, compared with monocytes pulsed in the absence of cytokines. This suppression was partly due to the secretion of prostaglandin E2 (PGE2) and IL-10 by GM-CSF-treated monocytes, since the combined use of indomethacin and anti-IL-10 antibodies during GM-CSF incubation and antigen pulsing restored T cell growth to about 65% of control levels. As confirmed by culture supernatant transfer experiments, maximal inhibition of T cell stimulation was also dependent on the direct contact between the T cells and GM-CSF-treated monocytes during antigen presentation. Collectively, these results imply that GM-CSF can either inhibit or enhance the re-stimulation of primed T cells by antigen-presenting monocytes or MoDC, respectively. PMID- 10844520 TI - Change in perforin-positive peripheral blood lymphocyte (PBL) subpopulations following exercise. AB - Perforin, one of the cytotoxic proteins of the immune system, plays a prominent role in protection against viral and bacterial infections. We investigated its expression in PBL and their CD3+, CD4+, CD8+ and CD16+ and/or CD56+ subpopulations in endurance athletes before and after a triathlon. Lymphocyte subpopulations were analysed by flow cytometry following separation of peripheral blood mononuclear cells and staining with antibodies against specific membrane antigens and intracellular perforin. The number of total lymphocytes decreased from 2.1 x 10(3)/microl before the triathlon to 1.0 x 10(3)/microl 1 h after the triathlon (P < 0.01). Interestingly, there was already a significant spontaneous decline in the percentage of CD3+/perforin+, and in CD8+/perforin+ cells, in the week proceeding the triathlon, when subjects were instructed to refrain from strenuous exercise training. The percentage of CD3+/perforin+, CD8+/perforin+ and CD16+ and/or CD56+/perforin+ cells in each lymphocyte subpopulation decreased 1 h after exercise even further from 14.3% to 5.8% (P < 0.05), 18.5% to 6.5% (P < 0.05) and 77.3% to 67.3%, respectively. However, at 18 h and 48 h after exercise the percentage of perforin-expressing CD3+, CD8+ and CD16+/56+ cells increased again towards baseline levels. Compared with normal controls, baseline perforin co-expression in CD3+ and CD8+ lymphocytes was significantly higher in trained athletes. From our data we conclude that trained athletes have an increased percentage of perforin+ PBL and that following exercise the percentage of perforin+ and therefore potentially cytotoxic lymphocytes transiently decreases in peripheral blood. PMID- 10844522 TI - Inhibition of cyclin A gene expression in human B cells by an immunosuppressant mizoribine. AB - Mizoribine has been shown to have beneficial effects in the treatment of rheumatoid arthritis and lupus nephritis, in which abnormal B cell functions are involved. Previous studies demonstrated that mizoribine directly suppresses the function of human B cells. The current study explored in detail the mechanism of the suppression of human B cell responses by mizoribine at the molecular level. Highly purified peripheral blood B cells obtained from normal healthy individuals were stimulated with Staphylococcus aureus Cowan I (SAC) plus IL-2 in the presence or absence of mizoribine or methotrexate for 48 h to 72 h. The expression of cyclin A mRNA was determined by semiquantitative reverse transcriptase-polymerase chain reaction followed by Southern hybridization. Although at pharmacologically attainable concentrations both mizoribine and methotrexate suppressed the production of IgM of SAC-activated B cells, mizoribine, but not methotrexate, decreased the expression of cyclin A protein as well as mRNA in B cells stimulated with SAC + IL-2. Of note, mizoribine facilitated the degradation of cyclin A mRNA in the presence of actinomycin D, indicating that mizoribine shortens the stability of cyclin A mRNA. The results indicate that mizoribine suppresses the expression of cyclin A mRNA in human B cells by down-regulating its stability, and thus down-regulates their responses. PMID- 10844523 TI - Human T cells with a type-2 cytokine profile are resistant to apoptosis induced by primary activation: consequences for immunopathogenesis. AB - The mechanisms leading to a relative dominance of T cells producing type 2 cytokines in certain human immune disorders are still unclear. We investigated the relative susceptibility to apoptosis induced by primary in vitro activation of human type 1 (producing interferon-gamma (IFN-gamma)) or type 2 (producing IL 4) T cells. Peripheral blood lymphocytes were isolated from patients with immune disorders characterized by expansion of type 2 cells (four with AIDS and hyper IgE/hypereosinophilia, one with Churg-Strauss syndrome, and one with idiopathic hypereosinophilic syndrome) or from individuals with normal cytokine balances. Cells were stimulated for 16 h with ionomycin and phorbol ester, and apoptosis of cytokine-producing cells was assessed by flow cytometry. T cells with a type-2 cytokine profile, i.e. producing IL-4 alone, were significantly more resistant to activation-induced apoptosis than those producing IFN-gamma alone. This was observed in AIDS patients, whose type 2 cells were mostly CD8+, as well as in the patients with Churg-Strauss and with hypereosinophilic syndrome. CD4+ and CD8+ IL 4-producing cells were equally resistant to apoptosis. Lower susceptibility to apoptosis of type-2 T cells was also observed in subjects with normal cytokine balances. Bcl-2 expression was high in type-2 cells and in viable type-1 cells, whereas it was low in apoptotic type-1 cells. Resistance to activation-induced apoptosis may explain the expansion of cells producing type-2 cytokines in certain immune disorders. PMID- 10844524 TI - Prevaccination with SRL172 (heat-killed Mycobacterium vaccae) inhibits experimental periodontal disease in Wistar rats. AB - Periodontal disease is a bacterial dental plaque-induced destructive inflammatory condition of the tooth-supporting tissues, which is thought to be mediated by T lymphocytes secreting T helper 2 (Th2) cytokines, resulting in recruitment of high numbers of antibody-producing B lymphocytes/plasma cells as well as polymorphonuclear leucocytes (PMN) secreting tissue-destructive components, such at matrix metalloproteinases and reactive oxygen metabolites into the gingival connective tissues. One treatment strategy may be to down-regulate the Th2 response to those dental plaque microorganisms which induce the destructive inflammatory response. In this study we have examined the effects of a potent down-regulator of Th2 responses on ligature-induced periodontal disease in an experimental rat model. A single s.c. injection into Wistar rats of 0.1 or 1 mg of SRL172, a preparation of heat-killed Mycobacterium vaccae (NCTC 11659), 13 days before application of the ligature, significantly reduced the subsequent destruction of the tooth-supporting tissues, as measured by loss of periodontal attachment fibres (P < 0.001) and bone (P < 0.002). This protective effect occurred not only on the experimental (ligatured) side but also on the control unligatured side. SRL172 has undergone extensive toxicological studies and safety assessments in humans, and it is suggested that it may provide a safe and novel therapeutic approach to periodontal disease. PMID- 10844525 TI - Co-stimulatory signals increase the reactivity of gammadelta T cells towards mycobacterial antigens. AB - Although it has been shown that gammadelta T lymphocytes are able to react with different cell-associated or soluble antigens, the immune repertoire of these cells appears to be skewed to the recognition of mycobacterial antigens. We have studied the number and reactivity of gammadelta T cells towards several mycobacterial antigens in patients with tuberculosis and leprosy, as well as their healthy contacts and control individuals. We found an increased number of Vdelta2+ cells in healthy contacts (PPD+ and lepromin+) and tuberculoid leprosy patients. The gammadelta T cells from lepromatous leprosy showed a decreased response to all antigens tested, but some of these patients exhibited a significant response to the 30-kD glycoprotein of Mycobacterium tuberculosis. Interestingly, the reactivity of gammadelta T cells against mycobacterial antigens was significantly increased by costimulatory signals generated through CD7, LFA-1, CD50 and CD69 in all groups. However, signalling through CD69 did not enhance the responsiveness of gammadelta lymphocytes from lepromatous patients. On the other hand, the in vitro blockade of IL-10 with a specific antibody enhanced the cell proliferation of gammadelta lymphocytes from lepromatous leprosy patients, whereas exogenous IL-10 had an opposite effect in most individuals studied. These results suggest the potential role of different cell membrane receptors in the regulation of gammadelta T cell proliferation induced by mycobacteria, as well as the possible involvement of IL-10 in this phenomenon. PMID- 10844526 TI - Protective effect of DNA immunization against mycobacterial infection is associated with the early emergence of interferon-gamma (IFN-gamma)-secreting lymphocytes. AB - The development of more effective anti-tuberculosis (TB) vaccines would contribute to the global control of TB. Understanding the activated/memory T cell response to mycobacterial infection and identifying immunological correlates of protective immunity will facilitate the design and assessment of new candidate vaccines. Therefore, we investigated the kinetics of the CD4+ T cell response and IFN-gamma production in an intravenous challenge model of Mycobacterium bovis bacille Calmette-Guerin (BCG) before and after DNA immunization. Activated/memory CD4+ T cells, defined as CD44hiCD45RBlo, expanded following infection, peaking at 3-4 weeks, and decreased as the bacterial load fell. Activated/memory CD4+ T cells were the major source of IFN-gamma and the level of antigen-specific IFN gamma-secreting lymphocytes, detected by ELISPOT, paralleled the changes in bacterial load. To examine the effects of a DNA vaccine, we immunized mice with a plasmid expressing the mycobacterial secreted antigen 85B (Ag85B). This led to a significant reduction in mycobacteria in the liver, spleen and lung. This protective effect was associated with the rapid emergence of antigen-specific IFN gamma-secreting lymphocytes which were detected earlier, at day 4, and at higher levels than in infected animals immunized with a control vector. This early and increased response of IFN-gamma-secreting T cells may serve as a correlate of protective immunity for anti-TB vaccines. PMID- 10844527 TI - Elevated serum concentrations of soluble CD14 in HIV- and HIV+ patients with tuberculosis in Africa: prolonged elevation during anti-tuberculosis treatment. AB - Data are limited regarding serum concentrations of soluble CD14 (sCD14), a marker of macrophage activation, in patients with active tuberculosis (TB) and during drug treatment. In this study, concentrations of sCD14 were measured in serum samples obtained from 105 African subjects who were categorized into one of four groups: persons with pulmonary TB alone (TB+HIV-, n = 30), pulmonary TB and HIV co-infection (TB+HIV+, n = 20), or HIV infection alone (TB-HIV+, n = 25), and healthy controls (TB-HIV-, n = 30). Mean total sCD14 was significantly increased in serum of patients with newly diagnosed pulmonary TB (mean = 6.6 g/ml, s.d. = 1.6 g/ml) compared with healthy controls (mean = 3.1 g/ml, s.d. = 0.6 g/ml; P < 0.0001), and this elevation comprised proportionate increases in the alpha (2.1 fold greater, P < 0.0001) and beta (2.0-fold greater, P < 0. 0001) forms of sCD14. Total sCD14 was also increased in serum of HIV-infected patients (mean = 4.1 g/ml, s.d. = 1.9 g/ml; P < 0.01), but the highest concentrations were observed in patients with pulmonary TB and HIV co-infection (mean = 8.7 g/ml, s.d. = 3.1 g/ml; P < 0.0001). Analysis of serum samples prospectively collected from TB+HIV-patients during the first 3 months of successful anti-TB treatment demonstrated steep reductions in mean concentrations of the acute-phase protein, C-reactive protein, and the soluble lymphocyte activation marker, sCD25. In contrast, levels of sCD14 increased during the first month of treatment and slowly declined thereafter. These data indicate that the serum concentration of sCD14 is not a sensitive index of response to anti-TB treatment and suggest that cellular activation resolves more slowly in the macrophage pool compared with the lymphocyte pool during anti-TB treatment. PMID- 10844528 TI - Up-regulation of human T lymphotropic virus type 1 (HTLV-1) tax/rex mRNA in infected lung tissues. AB - HTLV-1 has been implicated in certain pulmonary diseases. We previously demonstrated that expression of HTLV-1 tax/rex mRNA, encoding the transcriptional transactivator Tax, was closely associated with infiltration of activated T lymphocytes into lung tissue. To explore mechanisms of tax/rex expression in the lung, tax/rex mRNA expression and proviral DNA load were compared between peripheral blood mononuclear cells (PBMC) and bronchoalveolar lavage cells (BALC) from four patients with HTLV-1-associated myelopathy (HAM/TSP) and 13 carriers with various pulmonary symptoms. Semiquantitative detection of tax/rex mRNA strongly suggested that the lung was a preferential site for its expression. Proviral DNA loads in non-HAM/TSP carriers were variable but correlated well between PBMC and BALC in each individual, and revealed no relationship with tax/rex mRNA expression. In contrast, both cell groups from four HAM/TSP patients expressed detectable tax/rex mRNA accompanied by high proviral DNA load. The ratio of tax/rex mRNA expression to proviral DNA load was higher in BALC than in PBMC in three of four carriers and in three of four HAM/TSP patients, suggesting up-regulation of tax/rex mRNA in infected lung tissue. To analyse differences in distribution of HTLV-1 quasispecies between the two tissues, phylogenetic analysis was performed for sequence sets of the proviral tax open reading frame (ORF: 1059 bp) derived from PBMC and BALC of two infected individuals. Sequences derived from the two tissues distributed similarly to branches of phylogenetic trees, and there was no evidence of selective distribution of certain quasispecies in the lung. Our results suggest the presence of tissue-specific conditions that activate viral expression in infected cells in the lung. Constitutive exposure of this tissue to foreign antigens leading to up-regulation of basal viral promoter activity is likely to be one such mechanism. PMID- 10844529 TI - Absence of platelet CD40L identifies patients with X-linked hyper IgM syndrome. AB - CD40 ligand (CD40L), a membrane protein expressed on activated T cells, plays a pivotal role in B cell proliferation and differentiation. Mutations in the CD40L gene are associated with a rare immunodeficiency state, X-linked hyper IgM syndrome (XLHIGM). Recently, platelets have been described as capable of expressing CD40L within minutes of stimulation. We have developed a rapid technique to determine expression of CD40L on activated platelets by flow cytometry in whole blood. We have demonstrated that this technique is useful in neonatal screening, in rapid diagnosis and in determining reconstitution by donor bone marrow post-transplantation. PMID- 10844530 TI - Enhanced apoptosis of T cells in common variable immunodeficiency (CVID): role of defective CD28 co-stimulation. AB - CVID is a primary immune disorder in which hypogammaglobulinaemia may be associated with a number of T cell defects including lymphopenia, anergy, impaired lymphocyte proliferation and deficient cytokine secretion. In this study we show that T cells of CVID subjects, in comparison with control T cells, undergo spontaneous apoptosis in culture and markedly accelerated apoptosis after gamma-irradiation. Although costimulation of the CD28 receptor following engagement of the TCR/CD3 receptor normally provides a second signal necessary for IL-2 secretion, CD28 costimulation in CVID does not significantly increase IL 2 production, nor does this combination of activators enhance the survival of irradiated CVID T cells, as it does for cultured normal T cells. Addition of IL-2 enhances CVID T cell survival, suggesting that the IL-2 signalling pathways are normal. CVID T cells have similar expression of Bcl-2 to control T cells. CD3 stimulation up-regulates T cell expression of bcl-xL mRNA for normal T cells, but anti-CD28 does not augment bcl-xL expression for CVID subjects with accelerated apoptosis. Defects of the CD28 receptor pathway, leading to cytokine deprivation and dysregulation of bcl-xL, could lead to poor T cell viability and some of the cellular defects observed in CVID. PMID- 10844531 TI - Detection of Bruton's tyrosine kinase mutations in hypogammaglobulinaemic males registered as common variable immunodeficiency (CVID) in the Japanese Immunodeficiency Registry. AB - CVID is frequently diagnosed in male and female individuals with hypogammaglobulinaemia of unknown aetiology. To examine the possibility that sporadic male cases with X-linked agammaglobulinaemia (XLA), which is caused by mutations in the Bruton's tyrosine kinase (Btk) gene, might be misregistered as having CVID, we employed a flow cytometric test to identify XLA in hypogammaglobulinaemic males registered as CVID in the Japanese Immunodeficiency Registry. From 30 male cases registered as having CVID between 1992 and 1998, we selected 21 males with low or unreported peripheral B cell counts. Blood samples could be obtained from 11 patients and their mothers. Using flow cytometric analysis, the Btk-deficient status in monocytes was demonstrated in seven out of nine cases with decreased numbers of peripheral B cells. The diagnosis of XLA was confirmed in each of the seven patients by demonstration of Btk gene mutations in the patients or cellular mosaicism in the mother. This study demonstrates misregistration of XLA as CVID. PMID- 10844532 TI - Cytokine dysregulation in activated cystic fibrosis (CF) peripheral lymphocytes. AB - Recent studies demonstrate in vivo and in vitro cytokine dysregulation in CF epithelial cells. To see if these abnormalities may be generalized to other cells expressing cystic fibrosis transmembrane conductance regulator (CFTR) but not directly exposed to local inflammation, we studied mRNA transcription, intracellular protein production and extracellular secretion of IL-2, IL-4, IL-5, IL-10 and interferon-gamma (IFN-gamma) from freshly isolated blood mononuclear and CD4+ T cells from CF patients and controls. Cells were activated by phorbol myristate acetate (PMA) and anti-CD3, PMA-ionomycin, or lipopolysaccharide (LPS) and assessed for cytokine mRNA transcription by semiquantitative reverse transcriptase-polymerase chain reaction, intracellular protein production by flow cytometry, and secretion by supernatant ELISA. Cytokine expression was highly stimulus-dependent. CF cells showed higher IL-10 transcription than control cells after maximal activation by LPS (P = 0.01); despite this, cytokine production and secretion were equivalent to controls. CF cells showed lower cellular IL-10 production after PMA-anti-CD3 activation (P = 0.002). CF cells secreted less IFN gamma than control cells after maximal activation by PMA-anti-CD3 (1836 +/- 273 pg/ml versus 9635 +/- 3437 pg/ml, P = 0.04). IL-2, IL-4 and IL-5 regulation was similar to controls. We conclude that CF mononuclear cells show selective cytokine dysregulation after maximal activation, namely reduced IFN-gamma secretion and increased IL-10 mRNA without increased production or secretion. These findings extend defects described in respiratory epithelial cells to circulating immunoregulatory cells, suggesting a link between CF genotype and cytokine dysregulation. PMID- 10844533 TI - Suppression of ongoing experimental allergic encephalomyelitis (EAE) in Lewis rats: synergistic effects of myelin basic protein (MBP) peptide 68-86 and IL-4. AB - Mucosal myelin autoantigen administration effectively prevented EAE, but mostly failed to treat ongoing EAE. Patients with multiple sclerosis (MS), for which EAE is considered an animal model, did not benefit from oral treatment with bovine myelin. We anticipated that autoantigen, administered together with a cytokine that counteracts Th1 cell responses, might ameliorate Th1-driven autoimmune disease, and that nasal administration might considerably reduce the amounts of antigen + cytokine needed for treatment purposes. Lewis rats with EAE actively induced with myelin basic protein peptide (MBP 68-86) and Freund's complete adjuvant (FCA), received from day 7 post-immunization, i.e. after T cell priming had occurred, 120 microg MBP 68-86 + 100 ng IL-4 per rat per day for 5 consecutive days. These rats showed later onset, lower clinical scores, less body weight loss and shorter EAE duration compared with rats receiving MBP 68-86 or IL 4 only, or PBS. EAE amelioration was associated with decreased infiltration of ED1+ macrophages and CD4+ T cells within the central nervous system, and with decreased interferon-gamma (IFN-gamma) and tumour necrosis factor-alpha (TNF alpha) and enhanced IL-4, IL-10 and transforming growth factor-beta (TGF-beta) responses by lymph node cells. Simultaneous administration of encephalitogenic peptide + IL-4 by the nasal route thus suppressed ongoing EAE and induced IL-4, IL-10 and TGF-beta-related regulatory elements. PMID- 10844534 TI - CD8+ cells suppress oil-induced arthritis. AB - Oil-induced arthritis is a genetically restricted polyarthritis that develops in the DA rat after injection of the mineral oil Freund's incomplete adjuvant. Here, we investigated the role of the potentially disease-limiting cell populations CD8+ T cells, gammadelta T cells, natural killer (NK) cells and NK T cells in inguinal lymph nodes for the development of this adjuvant-induced arthritis. Flow cytometry analysis before and at disease onset revealed a higher proportion of lymph node T cells expressing NKR-P1 in the disease-resistant LEW.1AV1 compared with the disease-susceptible DA strain, suggesting that NK T cells might be disease protective. However, prophylactic in vivo administration of an anti-NKR P1 MoAb (clone 10/78) did not consistently affect the disease course. The proportion of CD8+ T cells and the ratio CD4+/CD8+ T cells in inguinal lymph nodes did not differ significantly between DA and LEW.1AV1 rats before or at disease onset. Nevertheless, prophylactic in vivo depletion of CD8+ cells by the OX8 MoAb in the DA strain resulted in an earlier disease onset compared with the control group, demonstrating that CD8+ cells regulate arthritis development. In vivo depletion of gammadelta T cells by the V65 MoAb did not alter the disease course, indicating that the disease-suppressive CD8+ cells are alphabeta T cells or NK cells. PMID- 10844536 TI - News from the european academy of andrology addendum to the supplement of the international journal of andrology, volume 22, supplement 1, 1999 PMID- 10844535 TI - Anti-beta2-glycoprotein I (GPI) autoantibodies, annexin V binding and the anti phospholipid syndrome. AB - We examined the role of autoantibodies to beta2-GPI and prothrombin (PT) in the inhibition of annexin V binding to cardiolipin (CL) and the association with clinical manifestations of the anti-phospholipid syndrome (APS). Plasma samples from 59 patients with anti-phospholipid (aPL) antibodies were studied. Affinity purification of total IgG and IgG anti-ss2-GPI antibodies was performed using staphylococcal protein A and phospholipid liposomes. Annexin V binding to CL was significantly inhibited by 31/59 (53%) aPL+ plasma samples. There was a significant association between annexin V inhibition and elevated levels of IgG anti-cardiolipin (aCL) (r = -0.62; P < 0.001), IgG anti-ss2-GPI (r = -0.67; P < 0. 001) and a weaker association with lupus anti-coagulant (r = -0.27; P = 0.05). There was no association with other isotypes of aCL and anti-ss2-GPI or with anti PT of any isotype. In patients with clinical manifestations of the APS there were higher levels of IgG aCL (median (range) Z score): 10.0 (0-17.6) versus 5.0 (0 16.1); P = 0.03), IgG anti-ss2-GPI (4.5 (0-11.3) versus 0.9 (0-9.7); P = 0.02) and greater inhibition of annexin V binding to CL (-3.4 (-11.4-0.6) versus -1.1 ( 10.8-1.2); P = 0.22). Odds ratios for the laboratory assays and the presence of clinical manifestations of the APS varied between 0.38 and 4.16, with the highest values for IgG aCL (4.16), IgG anti-ss2-GPI (3.28) and annexin V inhibition (2.85). Additional experiments with affinity-purified IgG antibodies indicated that inhibition of annexin V binding was dependent upon the concentration of ss2 GPI and anti-ss2-GPI antibodies. These results indicate that inhibition of annexin V binding to procoagulant phospholipid surfaces is dependent upon anti ss2-GPI antibodies and suggest a role for annexin V in the pathogenesis of the APS. PMID- 10844537 TI - Paternal transmission of genetic damage: findings in animals and humans. AB - The concept that mutations can be induced in the male germ-line and result in adverse effects in the offspring has achieved only limited acceptance despite considerable theoretical appeal. This is partly because fetal malformations are generally perceived to be induced solely as a result of maternally mediated events during gestation and partly because the low incidence of the end-points concerned make experimental approaches costly and time-consuming. Nonetheless, a substantial body of work relating to the hypothesis has accumulated in the last 20 years, which has never been reviewed in its entirety. A consideration of the available evidence indicates that preconceptional paternal exposure to mutagens (particularly radiation, cyclophosphamide and ethylnitrosourea) can indeed, under certain conditions, have adverse effects on offspring. The results suggest two principal mechanisms by which such effects may be induced: the induction of germ line genomic instability or the suppression of germ cell apoptosis. PMID- 10844538 TI - Regulation of inhibin production in the human male and its clinical applications. AB - Investigation of the regulation of testicular function has been enhanced by the ability to measure the dimeric, biologically active form of inhibin in men, inhibin B. This has allowed the demonstration that inhibin B is the afferent arm of the feedback loop from the testis that regulates FSH secretion, and investigation of inhibin B levels during reproductive development and in a variety of physiological and pathological states. Such studies have demonstrated many primary aspects of the relationship between FSH, testicular function (and in particular spermatogenesis) and inhibin B levels in blood, though many more specific questions remain. These include the precise nature of the relationship between inhibin B secretion and Sertoli cell function and how this relationship is influenced by the germ cell types present in the testis and by overall sperm production. When such information becomes available it will allow more accurate interpretation of blood concentrations of inhibin B. Similarly, apical secretion of inhibin B into seminal plasma may have considerable value in aiding assessment of the status of the seminiferous epithelium. Finally, neonatal secretion of inhibin B as a measure of Sertoli cell number and/or as a predictor of adult reproductive function offers novel possibilities for assessment and intervention. PMID- 10844539 TI - Chlorinated pesticides and heavy metals in human semen. AB - The concentration of chlorinated pesticides and heavy metals (lead and cadmium) was measured using gas liquid chromatography and the graphite tube atomizer of atomic absorption spectrophotometer, respectively, in semen samples collected from men in the normal human population. Significant concentrations of lead and cadmium were detected. Significant amounts of hexachlorocyclohexane (HCH) and its isomers alpha, beta, gamma and delta, the dichlorodiphenyl trichloroethane (DDT) metabolite 1,1,1-trichloro-2,2-bis (p chlorophenyl ethane) (pp'-DDE) and low values of 1,1,2-dichloro-2, 2-bis(p chlorophenyl ethane) (pp'-DDD) aldrin or endosulfan were detected. The presence of these xenobiotics in human semen might be related to the extensive use of pesticides, emission of exhaust from motor vehicles, consumption of tobacco and industrial operations. PMID- 10844540 TI - Sperm antigens recognized by antisperm antibodies present in sera of infertile adults and prepubertal boys with testicular failure. AB - Immunoblotting of a repertoire of sperm antigens reacting with antisperm antibodies present in sera of infertile adults and prepubertal boys with testicular failure was performed. In the subgroups selected for this study, 55% of examined infertile women, 65% of infertile men and 64% of prepubertal boys with gonadal failure gave positive results by Western blotting with extracted sperm antigens. Sperm antigens with molecular weights of 57, 58, 62, 63 and 66 kDa were the most immunodominant entities recognized by antisperm antibodies from prepubertal boys. No positive reactions were detected by Western blotting in a control population of fertile adults, whereas in a group of prepubertal healthy boys only one sample revealed reactivity against sperm antigens of 58 and 70 kDa. PMID- 10844541 TI - Thyroidectomy modulates rat prostatic monosaccharides. AB - Earlier studies have shown that thyroid hormones play an important role in glycoprotein metabolism in the prostate. In this paper, hypothyroidism induced changes in prostatic glycoprotein-associated monosaccharides and fructose concentrations were studied in 30-day-old peripubertal rats. Hypothyroidism was induced by total surgical thyroidectomy and confirmed by radioimmunoassay (RIA) of thyroid hormones and thyroid stimulating hormone (TSH). Animals were killed after 30 and 60 days post-thyroidectomy. To test the direct effect of thyroid hormones on prostatic monosaccharides, prostatic tissue was incubated for 24 h at 37 degrees C with 25, 50 or 100 ng/mL T3. Ventral prostatic hexosamines and sialic acid concentrations were significantly decreased in hypothyroid rats aged 30 and 60 days, whereas fucose concentration was increased at 30 days but decreased at 60 days in hypothyroid rats. In the case of the dorsolateral prostate, hypothyroidism enhanced the concentration of hexosamines but caused a decrease in levels of fucose, sialic acid and fructose, irrespective of the duration of hypothyroidism. Concentrations of fucose, fructose and hexosamines in the anterior prostate were decreased and sialic acid concentration was increased in 60-day hypothyroid rats. In in vitro studies, the concentrations of fucose, sialic acid and hexosamines were significantly increased in all three prostatic lobes irrespective of the T3 dose with which they had been incubated. A consistent increase in fructose concentration was observed in anterior and dorsolateral prostates when incubated with different doses of T3 stimulation. It is concluded that the influence of hypothyroidism on prostatic monosaccharides seems to be specific for each sugar in different lobes of the prostate. These results clearly show that thyroid hormones differentially regulate prostatic glycoprotein metabolism. PMID- 10844542 TI - Involvement of the calcium-specific protease, calpain, in the fertilizing capacity of human spermatozoa. AB - We recently reported the novel finding that human spermatozoa contain the calcium (Ca2+)-dependent protease, calpain. In somatic cells this protease mediates several cellular activities regulated by Ca2+ including membrane fusibility during cell-to-cell interactions. In this paper we examined the participation of sperm calpain in sperm-oocyte penetration, a process that is dependent on Ca2+ and involves membrane fusion between the two cells. Oocyte penetration was assessed using ejaculated spermatozoa from fertile men and zona-free hamster oocytes. Penetration rate was impaired by the presence of the active-site calpain inhibitor, calpain inhibitor-I, in a dose-dependent manner. At 1 mM, penetration scores were reduced by 65% (p < 0.01; n=5). The effects did not involve the oocyte, nor did the inhibitor alter sperm motility. Similar inhibitory effects on sperm penetration capacity were observed with specific antibodies directed either against calpain-I or calpain-II, the two forms of calpains described in somatic cells. At 1:1000 antibody dilution, penetration was inhibited 50 and 60% with anti-calpain-I and anti-calpain-II antibodies, respectively (p < 0.01; n=6). Furthermore, a combination of these two antibodies reduced the penetration rates by 75% (p < 0.01; n=6). We conclude that calpain inhibitor and anti-calpain antibodies impair human sperm capacity to fuse and penetrate the oocyte. These findings suggest that sperm calpain is a novel component of the biochemical processes that regulate the fertilizing capacity of human spermatozoa. PMID- 10844543 TI - Proteasomes in human spermatozoa. AB - In the present study we describe the localization of proteasomes in human spermatozoa by means of immunolabelling with different monoclonal and polyclonal antibodies detected by confocal microscopy. Western blotting confirmed the specificity of the antibodies and has shown that proteasomes are present in spermatozoa and in seminal fluid. In spermatozoa proteasomes are concentrated in the neck region where the centrioles are located. Some labelling was also detected at the periphery of the head, but no proteasomal antigens were detected in either the nucleus or associated with the flagellum. Proteasome inhibitors did not affect the motility of the spermatozoa, acrosome reaction nor zona binding. It is hypothesized that paternal proteasomes enter the oocyte during fertilization in tight association with the centrioles and may serve a special function during further development which can be associated with the function of a hypothetical proteolysis centre. PMID- 10844544 TI - Escherichia coli-induced alterations of human spermatozoa. An electron microscopy analysis. AB - This study evaluated if the negative influence of Escherichia coli on the motility of human spermatozoa is a consequence of E. coli-induced ultrastructural alterations. Suspensions of spermatozoa were artificially infected with E. coli from a serotyped, pathogenic strain and incubated at 37 degrees C for 6 h. After incubation, spermatozoa were fixed in glutaraldehyde, stained with osmium tetroxide and ruthenium red and embedded in Spurr(R)-resin followed by ultramicrotomy. The sections were analysed subsequently by use of transmission electron microscopy. Uninfected suspensions of spermatozoa in medium and bacterial suspensions served as controls. Negative contrast technique was performed to facilitate visualization of ultrastructural details of the bacterial capsule after experimental exposure to spermatozoa. Electron microscopic evaluation revealed multiple and profound alterations in the ultrastructure of spermatozoa such as membrane defects and cytoplasmic vacuoles exclusively in spermatozoa of infected samples (> 90%). Morphological alterations involved all superficial structures of spermatozoa, in particular the plasma membrane of the mid-piece and neck as well as the inner and outer acrosomal membrane of the acrosome, indicating that morphological defects account for the immobilization of spermatozoa by E. coli. The results suggest that E. coli infection of ejaculates results in immobilization and impaired acrosomal function in human spermatozoa, findings that support the indication for antimicrobial chemotherapy in symptomatic and silent infections that affect the ejaculate. PMID- 10844545 TI - Reduction in libido and fertility of male rats by administration of the nitric oxide (NO) synthase inhibitor N-nitro-L-arginine methyl ester. AB - The role of nitric oxide (NO) in libido and fertility of male rats was investigated by administration of the NO synthase inhibitor N-nitro-L-arginine methyl ester (L-NAME) (25 or 50 mg/kg/day). L-NAME caused marked reduction of precoital sexual behaviour, and a failure of most rats to mount or ejaculate during the test interval. In most matings that were successful, recovered spermatozoa were present in normal numbers. In mating experiments, L-NAME profoundly reduced the fertility of male rats. In those animals that did succeed in mating, the quantal pregnancy and the number of implants were reduced. After cessation of treatment with L-NAME, the fertility parameters returned close to normal. The inactive stereoisomer, D-NAME, caused none of the above effects when administered to rats. The results suggest that NO is essential for the expression of normal libido and fertility in male rats. It is likely that NO is required both in the male reproductive tract and in the brain. PMID- 10844546 TI - In this issue PMID- 10844547 TI - Identification of a locus for disseminated superficial actinic porokeratosis at chromosome 12q23.2-24.1. AB - Disseminated superficial actinic porokeratosis is an autosomal dominant cutaneous disorder characterized by many uniformly small, minimal, annular, anhidrotic, and keratotic lesions. The genetic basis for this disease is unknown. Using a genomewide search in a large Chinese family, we identified a locus at chromosome 12q23.2-24. 1 responsible for disseminated superficial actinic porokeratosis. The fine mapping study indicates that the disseminated superficial actinic porokeratosis gene is located within a 9.6 cM region between markers D12S1727 and D12S1605, with a maximum two-point LOD score of 20.53 (theta = 0.00) at D12S78. This is the first locus identified for a genetic disease where the major phenotype is porokeratosis. The study provides a map location for isolation of a gene causing disseminated superficial actinic porokeratosis. PMID- 10844548 TI - Keratinocyte growth regulation in defined organotypic cultures through IL-1 induced keratinocyte growth factor expression in resting fibroblasts. AB - Balanced keratinocyte proliferation and differentiation resulting in regular tissue organization strictly depend on dermal support. Organotypic cultures represent biologically relevant in vitro models to study the molecular mechanism of the underlying dermal-epidermal interactions. To mimic the state of resting fibroblasts in the dermis, postmitotic (irradiated) fibroblasts were incorporated in the collagen matrix, where they typically support epidermal proliferation and tissue organization. In coculture with keratinocytes, fibroblasts exhibit an enhanced expression of keratinocyte growth factor and the interleukin-1 receptor (type I), which further increase with culture time. In cocultured keratinocytes, keratinocyte growth factor receptor as well as RNA expression and protein release of interleukin-1alpha and interleukin-1beta are upregulated. We hypothesized that the modulated cytokine expression represents a basic mechanism for keratinocyte growth regulation. The functional significance of this double paracrine pathway, i.e., induction of keratinocyte growth factor expression in fibroblasts by keratinocytes via release of interleukin-1, was confirmed by interfering with both signaling elements: (i) interleukin-1-neutralizing antibodies and interleukin-1 receptor antagonist significantly inhibited keratinocyte growth factor release, keratinocyte proliferation, and tissue formation comparable to the effect produced by keratinocyte-growth-factor-blocking antibodies; (ii) addition of keratinocyte growth factor to cocultures with inactivated interleukin 1 pathway completely reverted growth inhibition; (iii) in organotypic cocultures with subthreshold fibroblast numbers both interleukin-1 and keratinocyte growth factor restored the impaired epidermal morphogenesis. Thus, epidermal tissue regeneration in organotypic cocultures is mainly regulated by keratinocyte derived interleukin-1 signaling, which induces keratinocyte growth factor expression in cocultured fibroblasts. This demonstrates a novel role for interleukin-1 in skin homeostasis substantiating data from wound healing studies in vivo. PMID- 10844549 TI - Allergens and irritants transcriptionally upregulate CD80 gene expression in human keratinocytes. AB - The human CD80 costimulatory molecule is an important signal between professional antigen-presenting cells and T helper cells. The immunobiology of CD80 expression by keratinocytes, especially during allergic and irritant contact dermatitis, however, is less well understood. CD80 cell surface expression and gene transcription by keratinocytes was increased when keratinocytes were exposed to certain allergens (chemicals that induce inflammation via hapten-specific T cells) and irritants (chemicals that are toxic to epidermal cells). Therefore, the human CD80 promoter was cloned and luciferase reporter constructs containing various promoter fragments were engineered. Promoter mapping of these CD80 constructs in transiently transfected keratinocytes showed that a construct containing the proximal 231 bp immediately upstream of the transcription start site of the CD80 promoter was most active in keratinocytes and was inducible to a level ranging from 2- to 10-fold higher in keratinocytes treated with certain allergens and irritants, compared with untreated keratinocytes. This pattern of promoter fragment activity in keratinocytes is identical to that found in professional antigen-presenting cells. This is the first demonstration that the CD80 promoter is active in keratinocytes and that this activity is further increased in keratinocytes treated with certain allergens and irritants. These data suggest that allergens and irritants may, in part, break peripheral tolerance by their direct effects on keratinocyte costimulatory molecule expression, thereby facilitating interactions with epidermotropic T helper cells via the CD80-CD28 or CTLA-4 pathways. PMID- 10844550 TI - The neurofibromatosis type 1 (Nf1) tumor suppressor is a modifier of carcinogen induced pigmentation and papilloma formation in C57BL/6 mice. AB - There is increasing evidence implicating the human NF1 gene in epithelial carcinogenesis. To test if NF1 can play a part in skin tumor formation, we analyzed effects of the skin cancer initiator dimethylbenz-anthracene and/or the tumor promoter 12-O-tetradecanoyl-13-acetylphorbol on mice heterozygous for null mutations in Nf1 (Nf1+/-). Mice were on the C57BL/6 background, noted for resistance to chemical carcinogens. Nf1+/- mice (18 of 24) developed papillomas after treatment with dimethylbenzanthracene and 12-O-tetradecanoyl-13 acetylphorbol; papillomas did not develop in wild-type C57BL/6 mice nor Nf1+/- mice treated with 12-O-tetradecanoyl-13-acetylphorbol alone. All papillomas analyzed (six of six) had mutations in codon 61 of H-ras, demonstrating strong cooperation between the Nf1 GTPase activating protein for Ras, neurofibromin, and Ras-GTP. After exposure to 12-O-tetradecanoyl-13-acetylphorbol, Nf1+/- keratinocytes showed significant, sustained, increases in proliferation, implicating Nf1 in phorbol ester responsive pathways. Thus, Nf1 levels regulate the response of keratinocytes to 12-O-tetradecanoyl-13-acetylphorbol. Nf1+/- mice also showed a 2-fold increase in the development of pigmented skin patches stimulated by dimethylbenzanthracene; patches were characterized by hair follicles in anagen phase, implicating keratinocytes in the aberrant hyperpigmentation. Our results show that mutation in the Nf1 gene causes abnormal keratinocyte proliferation that can be revealed by environmental assaults such as carcinogen exposure. The data support a plausible role for NF1 mutation in human epithelial carcinogenesis. PMID- 10844551 TI - Keratin 17 expression in the hard epithelial context of the hair and nail, and its relevance for the pachyonychia congenita phenotype. AB - The hard-keratin-containing portion of the murine hair shaft displays a positive immunoreactivity with an antibody against the soft epithelial keratin, K17. The K17-expressing cell population is located in the medulla compartment of the hair. Consistent with this observation, K17-containing cells also occur in the presumptive medulla precursor cells located in the hair follicle matrix. Western blot analysis of hair extracts prepared from a number of mouse strains confirms this observation and suggests that K17 expression in the hair shaft is a general trait in this species. The expression of K17 in human hair extracts is restricted to eyebrow and facial hair samples. These are the major sites for the occurrence of the pili torti (twisted hair) phenotype in the type 2 (Jackson-Lawler) form of pachyonychia congenita, previously shown to arise from inherited K17 mutations. Given that all forms of pachyonychia congenita show an involvement of the nail, we compared the expression of the two other genes mutated in pachyonychia congenita diseases, K6 and K16, with that of K17 in human nail. All three keratins are abundantly expressed within the nail bed epithelium, whereas K17 protein is expressed in the nail matrix, which contains the epithelial cell precursors for the nail plate. Our data suggest a role for K17 in the formation and maintenance of various skin appendages and directly support the concept that pachyonychia congenita is a disease of the nail bed. PMID- 10844552 TI - Neurotrophin-4 production by human epidermal keratinocytes: increased expression in atopic dermatitis. AB - Chronic inflammatory conditions of human skin, such as prurigo lesions of atopic dermatitis, are characterized clinically by intense pruritus and histologically by increased innervation. Regulation of skin innervation is thought to depend on neurotrophic factors. In this study, human skin cells were identified as a source of neurotrophins. Cultured keratinocytes expressed neurotrophin-4, whereas dermal fibroblasts expressed neurotrophin-3. In vitro stimulation with interferon-gamma, a marker cytokine for atopic eczema, induced keratinocyte neurotrophin-4 production, which was able to support growth of a neuroglioblastoma-derived cell line. In vivo, immunohistochemistry of human skin for neurotrophins showed neurotrophin-4 staining in the epidermal layer and neurotrophin-3 staining in the dermal compartment. Neurotrophin-4 but not neurotrophin-3 expression was markedly increased in interferon-gamma-injected skin. Prurigo lesions of atopic dermatitis skin were characterized by intense epidermal staining for neurotrophin-4, suggesting a pathophysiologic role for this neurotrophin in the increased innervation characteristic for these skin lesions. This study demonstrates differential expression and regulation of neurotrophins in human skin. It also identifies keratinocyte-derived neurotrophin-4 as a possible link between the immune and the nerve system of human skin. PMID- 10844553 TI - Expression of human macrophage metalloelastase (MMP-12) by tumor cells in skin cancer. AB - Matrix metalloproteinases play an essential role in tumor growth and invasion. Different matrix metalloproteinases are often expressed in cancers with distinct patterns. To investigate the role of human macrophage metalloelastase (MMP-12) in epidermal tumors, we studied human macrophage metalloelastase mRNA and protein expression in malignant squamous cell and basal cell carcinomas, and in premalignant Bowen's disease. Human macrophage metalloelastase was detected in 11 of 17 squamous cell carcinomas in epithelial cancer cells, whereas macrophages were positive in 15 of 17 samples. In basal cell carcinomas, human macrophage metalloelastase was more often found in macrophages (seven of 19) than in cancer cells (four of 19). Human macrophage metalloelastase mRNA was also detected in three cell lines derived from squamous cell carcinomas of the head and neck and in transformed HaCaT cells, whereas premalignant tumors and primary keratinocytes were negative for human macrophage metalloelastase mRNA. Western analysis revealed human macrophage metalloelastase protein in squamous cell carcinoma cells. Our results show that human macrophage metalloelastase can be expressed in vivo and in vitro by transformed epithelial cells and indicate that the level of human macrophage metalloelastase expression correlates with epithelial dedifferentiation and histologic aggressiveness. PMID- 10844554 TI - An experimental model of cutaneous infection induced by superantigen-producing Staphylococcus aureus. AB - Skin infections caused by Staphylococcus aureus, such as erysipelas, are commonly occurring, painful, and costly for society. Despite the high prevalence of this condition, little is known about the host immune responsiveness and bacterial virulence factors during S. aureus dermatitis. We present here a mouse model of infectious dermatitis in which S. aureus is inoculated by an intracutaneous injection to the shaved back of NMRI mice. Visible skin inflammation, characterized by redness and swelling, was noted 48 h after inoculation of staphylococci in mice that received 2 x 108 colony-forming units of S. aureus. Microscopic evaluation revealed a dermal and subcutaneous infiltrate rich in macrophages and neutrophilic granulocytes already within 6 h after inoculation. A sparse influx of T lymphocytes was noted somewhat later. Bacterial cultures from skin revealed high numbers of staphylococci early after inoculation, with a successive decline during 2 wk follow-up. Total white blood cell count as well as the number of polymorphonuclear leukocytes peaked 2 d after bacterial inoculation. Also, serum interleukin-6 levels peaked within 2 d, with a 10-fold increase compared to non-infected control mice, indicating a systemic reaction to skin infection. The role of toxic shock syndrome toxin 1 in the pathogenesis of the dermatitis was assessed using isogenic S. aureus strains. Even though the gross inflammatory skin reaction was similar for mice infected with either of the strains, it was apparent that bacteria secreting toxic shock syndrome toxin 1 preferentially triggered influx of T lymphocytes to the skin. In addition, mice inoculated with staphylococci producing toxic shock syndrome toxin 1 showed a weight decrease during the experiment whereas mice inoculated with the isogenic strain showed a weight increase. This model of staphylococcal dermatitis will enable future in-depth studies regarding the host-bacterium relationship. PMID- 10844555 TI - Tetraspanins are localized at motility-related structures and involved in normal human keratinocyte wound healing migration. AB - We have described previously that beta1 integrins, which mediate keratinocyte cell adhesion and migration, are in ligand-occupied conformation at the basal surface but not at the lateral and apical surfaces of keratinocytes. This led us to study the cellular localization and function of tetraspanin molecules, which have been postulated to modulate integrin activity. We found that CD9 and CD81 are highly expressed by keratinocytes clearly delineating filopodia at lateral and apical surfaces. CD63 and CD151 are largely expressed in the intracellular compartment, although some membrane expression is observed. We found accumulation of CD9, CD81, and CD151 together with alpha3 and beta1 integrins at intercellular junctions. In low calcium medium, this intercellular space is crossed by a zipper of filopodia enriched in alpha3beta1 and tetraspanin proteins. Interestingly, the expression of CD9, CD81, and beta1 and alpha3 integrins was detected in the footprints and rippings of motile keratinocytes, suggesting their role in both adhesion to extracellular matrix and keratinocyte motility. beta1 integrins were only partially activated in the rips, whereas cytoskeleton-linking proteins such as talin were completely absent. On the other hand, antitetraspanin antibodies did not stain focal adhesions, which contain talin. The involvement of tetraspanins in keratinocyte motility was assessed in a wound healing migration assay. Inhibition of cell migration was observed with antibodies to CD9, CD81, beta1, and alpha3, and, to a lesser extent, to CD151. Together these results indicate that tetraspanin-integrin complexes might be involved in transient adhesion and integrin recycling during keratinocyte migration, as well as in intercellular recognition. PMID- 10844556 TI - A mutation in the V1 domain of K16 is responsible for unilateral palmoplantar verrucous nevus. AB - Palmoplantar keratodermas are a group of heterogeneous diseases characterized by thickening, and marked hyperkeratosis, of the epidermis of the palms and soles. Palmoplantar keratodermas can be divided into four major classes: diffuse, focal, punctate, and palmoplantar ectodermal dysplasias. All forms are genetic diseases inherited as autosomal dominant disorders. We studied a patient exhibiting a localized thickening of the skin in parts of the right palm and the right sole, following Blaschko's lines, that does not fit into any classes already described. We sequenced the keratin 16 cDNA derived from skin biopsy material from affected and non affected palms. The keratin 16 cDNA sequence from lesional epidermis showed a 12 base pair deletion (309-320del), which deletes codons 104-107. The mutation is predicted to delete four amino acids, GGFA, from the V1 domain of the keratin 16 polypeptide, close to the 1A domain. Full-length keratin 16 cDNA sequence derived from the unaffected palm was completely normal, consistent with a postzygotic mutation as is suggested by the mosaicism observed. We defined this new clinical entity, "unilateral palmoplantar verrucous nevus", rather than localized or focal epidermolytic palmoplantar keratodermas, as the lesions are present only on one side of the body and follow Blaschko's lines. This study is a report of a mosaic mutation in keratin 16 and also the association of a mutation in the V1 domain of a type I keratin associated with a human disease. PMID- 10844557 TI - Microanalysis of melanins in mammalian hair by alkaline hydrogen peroxide degradation: identification of a new structural marker of pheomelanins. AB - A highly sensitive, easy-to-perform method for melanin analysis in pigmented tissues based on alkaline hydrogen peroxide degradation has been developed and accomplishes simultaneous determination of eumelanins and pheomelanins. Pyrrole 2,3,5- tricarboxylic acid, the typical eumelanin marker, was obtained in higher yields than in previous procedures. A benzothiazole acid, 6-(2-amino-2 carboxyethyl)-2-carboxy-4-hydroxybenzothiazole, characterized in our previous studies as a specific marker of pheomelanins, and the newly identified 1,3 thiazole-2,4, 5-tricarboxylic acid were also used for pigment analysis. Optimal yields of the pigment markers were obtained at 24 h reaction time. Pyrrole-2,3,5 tricarboxylic acid, 6-(2-amino-2-carboxyethyl)-2-carboxy-4-hydroxybenzothiazole, and 1, 3-thiazole-2,4,5- tricarboxylic acid were quantified in a single chromatographic analysis without fractionation or work up of the degradation mixture. The linearity (linearity coefficient from 0.997 to 0.999) was excellent and the inter-assay (percentage coefficient of variation values in the range 0.2 2, n = 6) and intra-assay (percentage coefficient of variation values Phe) that differed significantly between patients and controls was identified (position 619). Owing to a high allele frequency in a larger control group, however, and an insignificant influence of the variant on the age at onset distribution curve based on a large psoriasis population, we could not confirm that this coding single nucleotide polymorphism was involved in disease etiology. We also examined the single nucleotide polymorphism in position 1243, recently proposed to have an influence on the pathogenesis of the disease. This polymorphism showed less association to the disease as compared with the single nucleotide polymorphism at positions 619 and 722. Such a high degree of variation present also in an HLA gene which is not involved in immune response indicates the difficulty involved in assessing the role of a specific allele in the pathogenesis of a complex disease in this region. A strong association effect due to linkage disequilibrium in an extended region in the HLA complex is also a complicating factor. PMID- 10844562 TI - 1alpha,25-dihydroxyvitamin D3 induced differentiation of cultured human keratinocytes is accompanied by a PKC-independent regulation of AP-1 DNA binding activity. AB - 1alpha,25(OH)2D3 and its analogs are potent mediators of keratinocyte differentiation in vitro. The precise mechanism of this action is still unknown. The nuclear transcription factor activator protein 1 seems to play an important role in keratinocyte differentiation. The purpose of this study was to investigate the effect of 1alpha,25(OH)2D3 on activator protein 1 DNA binding activity in cultured human keratinocytes. In a time-course study of human keratinocytes incubated with 1alpha,25(OH)2D3 (10-7-10-11 M) a significant dose dependent increase in activator protein 1 DNA binding activity as determined by electrophoretic mobility shift assay was seen after 36 h. This increase was followed by a significant dose-dependent decrease in activator protein 1 DNA binding activity after 72 h. When differentiation was induced by raising the calcium concentration in the culture medium from 0.09 to 0.3 mM a similar increase in activator protein 1 DNA binding activity was observed after incubation for 48 h. Pharmacologic down-modulation of the protein kinase C activity with GF 109203X reversed the calcium-induced increase in activator protein 1 DNA binding activity and abolished keratinocyte differentiation as determined by a transglutaminase assay. In contrast, activator protein 1 DNA binding activity and keratinocyte differentiation were not affected when protein kinase C activity was down-modulated in the experiments with 1alpha,25(OH)2D3. The activator protein 1 complex in human keratinocytes consists of dimers of Fra 1, Fra-2, c-Jun, JunD, and c-Fos. Our results demonstrate that 1alpha, 25(OH)2D3- and calcium-induced keratinocyte differentiation are accompanied by changes in activator protein 1 DNA binding activity. Protein kinase C activation appears to be essential for the calcium-dependent induction of keratinocyte differentiation, whereas a protein-kinase-C-independent activation of activator protein 1 DNA binding and keratinocyte differentiation is responsible for the 1alpha,25(OH)2D3 induced effects. PMID- 10844563 TI - Different transcriptional activity and in vitro TNF-alpha production in psoriasis patients carrying the TNF-alpha 238A promoter polymorphism. AB - Genes encoded on chromosome 6 within the major histocompatibility complex region are thought to play an important role in the pathogenesis of psoriasis. A potential candidate gene is tumor necrosis factor alpha. The tumor necrosis factor alpha promoter contains several polymorphisms including two G-->A transitions at position -308 and -238, which are the most common in Caucasian populations. The TNF238.2 (-238A) allele has been strongly associated with psoriasis. We have investigated the effect of the -238 and -308 variants on transcription of the tumor necrosis factor alpha gene in luciferase reporter gene assays. In addition, peripheral blood mononuclear cells of 47 patients with psoriasis and 43 controls were stimulated with different antigens and mitogens (streptococcal sonicate and superantigen, lipopolysaccharide, phorbol-12 myristate, phytohemagglutinin, CD3 antibodies) and tumor necrosis factor alpha production was measured in supernatants by enzyme-linked immunosorbent assay. The psoriasis-associated tumor necrosis factor alpha promoter allele TNF238.2 showed a significantly decreased transcriptional activity. Peripheral blood mononuclear cells carrying this allele produced significantly less tumor necrosis factor alpha after stimulation with T cell mitogens and streptococcal antigens in comparison to controls. The promoter allele TNF238.2 seems to influence tumor necrosis factor alpha production; a possible role in the pathogenesis of psoriasis has to be further evaluated. PMID- 10844564 TI - Hyaluronan exists in the normal stratum corneum. AB - Hyaluronan is well known to exist as a water-sorbed macromolecule in the extracellular matrix. We here examined whether hyaluronan exists in the normal stratum corneum. High performance liquid chromatography was used to quantify hyaluronan content in the stratum corneum, epidermis (including stratum corneum), and dermis of mice, with the resulting dry weights being 22.3 +/- 2.9, 15.1 +/- 1.5, and 738.6 +/- 31.6 microg per g, respectively. Normal mouse skin was then labeled with [3H]-glucosamine in an organ culture, and accumulation of [3H] labeled hyaluronan and its molecular mass were determined separately for the stratum corneum, epidermis, and dermis. In the stratum corneum, [3H]-labeled hyaluronan was accumulated linearly over the 3-d culture period. After the 3-d culture period, the epidermis synthesized twice the amount (expressed as dpm per mg dry weight) of [3H]-labeled hyaluronan as the dermis, whereas the stratum corneum and dermis showed nearly the same content of [3H]-labeled hyaluronan. The molecular mass of [3H]-labeled hyaluronan was highest (>1.0 x 106) in the dermis and clearly lower (<6.0 x 104) in the stratum corneum. Based on these results, we here confirm that hyaluronan is supplied from keratinocytes beneath the stratum corneum layer, and is present in the normal stratum corneum. We speculate that hyaluronan may play a role in moisturizing the stratum corneum and/or regulating its mechanical properties. PMID- 10844565 TI - Wild-type p53 gene transfer inhibits invasion and reduces matrix metalloproteinase-2 levels in p53-mutated human melanoma cells. AB - The tumor suppressor gene p53 has inhibitory effects on cell growth and angiogenesis and induces apoptosis when overexpressed in melanoma and in a variety of tumor cells by adenovirus-mediated gene transfer. The invasive ability of tumor cells, facilitating local infiltration and metastasis, is related to matrix metalloproteinase levels. In melanoma, matrix metalloproteinase-2 and matrix metalloproteinase-9 have a prominent role in this process. The aim of this study was to evaluate whether wild-type p53 overexpression, obtained by a recombinant adenovirus vector (AdCMV.p53), affects cell invasiveness through modulation of matrix metalloproteinase-2 and matrix metalloproteinase-9. Two human melanoma cell lines were used in this study: the SK-MEL-110, carrying a mutated p53 gene, and the SK-MEL-147, carrying the wild-type p53 gene. SK-MEL-110 cells infected with AdCMV.p53 exhibited decreased invasion capability from day 1 after infection, compared with cells not infected or infected with the control vector AdCMV.Null. This reduced invasiveness was associated with decreased matrix metalloproteinase-2 levels in conditioned media whereas no changes were detected in matrix metalloproteinase-9 secreted levels. No modulation in matrix metalloproteinase-2 mRNA levels was detectable, however, after wild-type p53 gene transfer. Furthermore, protein expression of secreted tissue inhibitor of metalloproteinase-2 was not altered by AdCMV.p53 treatment. In contrast, in SK MEL-147 cells, AdCMV.p53 did not affect cell invasiveness and levels of secreted matrix metalloproteinase-2. Gene transfer of wild-type p53 inhibited proliferation of both cell lines, showing that also SK-MEL-147 cells respond to wild-type p53 overexpression. This novel mechanism of action of wild-type p53 gene transfer may contribute to its antitumor effect by downregulating cell invasion and matrix metalloproteinase-2 secreted levels in mutated p53 human melanoma cell lines. PMID- 10844566 TI - Novel point mutations in the steroid sulfatase gene in patients with X-linked ichthyosis: transfection analysis using the mutated genes. AB - X-linked ichthyosis is caused by steroid sulfatase deficiency which results from abnormalities in its coding gene. The majority of X-linked ichthyosis patients ( approximately 90%) have complete or partial deletions of the steroid sulfatase gene. In this study, we examined the mutations of the steroid sulfatase gene in two unrelated X-linked ichthyosis patients without complete deletion of the gene. Polymerase chain reaction-single-strand conformation polymorphism and direct sequencing analyses showed that each patient has a different single base pair substitution within exon 8 encoding the C-terminal half of the steroid sulfatase polypeptide. Both mutations resulted in the transversion of functional amino acids: a G-->C substitution at nucleotide 1344, causing a predicted change of a glycine to an arginine, and a C-->T substitution at nucleotide 1371, causing a change from a glutamine to a stop codon. In vitro steroid sulfatase cDNA expression using site-directed mutagenesis revealed that these mutations are in fact pathogenic and reflect the levels of steroid sulfatase enzyme activities in each of the X-linked ichthyosis patients. PMID- 10844567 TI - Expression of activated N-ras in a primary melanoma cell line counteracts growth inhibition by transforming growth factor-beta. AB - One critical factor in melanoma progression is the change from radial growth phase to vertical growth phase. We previously showed a high incidence of ras mutations in progressing but not early human melanomas. We also found that stable expression of activated Ras in a primary human melanoma cell line (WM35) led to enhanced proliferation, anchorage-independent survival, migration and invasion in vitro and enhanced subcutaneous tumor formation in vivo, transforming the melanoma phenotype from the radial growth phase to the vertical growth phase. Inhibitory cytokines, especially transforming growth factor-beta, are important in homeostasis of normal human melanocytes. Proliferation of early melanoma cells can be inhibited by transforming growth factor-beta, whereas more aggressive stages lose this response. Using a transforming growth factor-beta activated luciferase reporter transiently transfected into WM35, WM35N-ras, and WM35H-ras (WM35 transfected with mutant N-ras or H-ras genes), we demonstrated significant decreases (p < 0. 04) in transforming growth factor-beta induced reporter expression in both ras transfected cell lines. Transforming growth factor-beta also induced significant decreases (p < 0.002) in the proportion of WM35 cells in S-phase of the cell cycle; this effect was not observed in WM35N-ras cells. Furthermore, we demonstrated that an important controlling factor in transforming growth factor-beta inhibition of cell cycle progression, the phosphorylation of the Rb protein, was altered in WM35N-ras; transforming growth factor-beta caused a marked relative increase in hypophosphorylated pRb in WM35 cells, but not in WM35N-ras. These data suggest that activated Ras plays an important part in melanoma progression from the radial growth phase to the vertical growth phase by counteracting inhibition by cytokines such as transforming growth factor-beta, thus providing a growth advantage. PMID- 10844568 TI - A role for Bax in the regulation of apoptosis in mouse mast cells. PMID- 10844569 TI - Calgranulin C is overexpressed in lesional psoriasis. PMID- 10844570 TI - VEGF and its receptors are expressed in a pediatric angiosarcoma in a patient with Aicardi's syndrome. PMID- 10844571 TI - Cloning of canine Dsg1 and evidence for alternative polyadenylation. PMID- 10844572 TI - Sex hormones, mood, mental state and memory PMID- 10844573 TI - Sex hormones, mood, mental state and memory PMID- 10844574 TI - Vasopressin regulation of noradrenaline release within the supraoptic nucleus. AB - The effect of electrically evoked dendritic vasopressin release on noradrenaline release into the hypothalamic supraoptic nucleus was assessed by in vivo microdialysis in conjunction with high pressure liquid chromatography and electrochemical detection. Electrical activation of magnocellular supraoptic neurones by stimulation of their axons at the level of the neural lobe significantly increased noradrenaline release into the nucleus (2.5-fold, P<0.03). This increase was completely blocked by administration of a nonpeptide vasopressin V1a receptor antagonist via the microdialysis probe. These data suggest that dendritically released vasopressin facilitates noradrenaline release into the hypothalamic nucleus. PMID- 10844575 TI - Glucocorticoids regulate pituitary growth hormone secretagogue receptor gene expression. AB - Glucocorticoids regulate growth hormone (GH) secretion by modulating both hypothalamic and pituitary function. At the level of the pituitary, glucocorticoids increase GH and GH-releasing hormone receptor (GHRH-R) gene expression. To test if glucocorticoids might also regulate the pituitary expression of the recently identified GH secretagogue (GHS) receptor, GHS-R; adult male rats were adrenalectomized or sham operated, and treated with the synthetic glucocorticoid (dexamethasone, 200 microg/day) or vehicle for 8 days. Pituitary GHS-R mRNA levels were assessed by reverse transcriptase polymerase chain reaction (RT-PCR). Adrenalectomy decreased pituitary GHS-R mRNA to 45% of vehicle-treated, sham-operated rats (P < 0.05). Administration of dexamethasone increased GHS-R mRNA levels in sham-operated as well as in adrenalectomized rats (199 +/- 24% (P < 0.05) and 369 +/- 48% (P < 0.01) of vehicle-treated controls). Addition of dexamethasone to primary rat pituitary cell cultures increased GHS-R mRNA levels in a dose- and time-dependent manner while the transcriptional inhibitor, actinomycin D, completely blocked the stimulatory action of dexamethasone. Taken together, these results suggest glucocorticoids directly increase pituitary GHS-R mRNA levels by stimulating GHS-R gene transcription. PMID- 10844576 TI - Chronic corticosterone administration dose-dependently modulates Abeta(1-42)- and NMDA-induced neurodegeneration in rat magnocellular nucleus basalis. AB - The impact of glucocorticoids on beta-amyloid(1-42) (Abeta(1-42)) and NMDA induced neurodegeneration was investigated in vivo. Abeta(1-42) or NMDA was injected into the cholinergic magnocellular nucleus basalis in adrenalectomized (ADX) rats, ADX rats supplemented with 25%, 100%, 2x100% corticosterone pellets, or sham-ADX controls. Abeta(1-42)- or NMDA-induced damage of cholinergic nucleus basalis neurones was assessed by quantitative acetylcholinesterase histochemistry. Plasma concentrations of corticosterone and cholinergic fibre loss after Abeta(1-42) or NMDA injection showed a clear U-shaped dose-response relationship. ADX and subsequent loss of serum corticosterone potentiated both the Abeta(1-42) and NMDA-induced neurodegeneration. ADX+25% corticosterone resulted in a 10-90 nM plasma corticosterone concentration, which significantly attenuated the Abeta(1-42) and NMDA neurotoxicity. ADX+100% corticosterone (corticosterone concentrations of 110-270 nM) potently decreased both Abeta(1-42) and NMDA-induced neurotoxic brain damage. In contrast, high corticosterone concentrations of 310-650 nM potentiated Abeta(1-42)- and NMDA-triggered neurodegeneration. In conclusion, chronic low or high corticosterone concentrations increase the vulnerability of cholinergic cells to neurotoxic insult, while slightly elevated corticosterone levels protect against neurotoxic injury. Enhanced neurotoxicity of NMDA in the presence of high concentrations of specific glucocorticoid receptor agonists suggests that the corticosterone effects are mediated by glucocorticoid receptors. PMID- 10844577 TI - Evidence suggesting that the potentiating action of neuropeptide Y on luteinizing hormone (LH)-releasing hormone-induced LH release remains unaltered in aged female rats. AB - Several lines of evidence suggest that one of the mechanisms by which the hypothalamic neuropeptide Y plays an obligatory role in the preovulatory luteinizing hormone (LH) discharge in young rats is to potentiate the action of LH-releasing hormone (LHRH) on LH release at the level of the pituitary. This study examined whether an alteration in the potentiating action of neuropeptide Y on LHRH-induced LH release may contribute to the attenuation or absence of LH surges during female reproductive ageing. Young regularly cycling (2-3-month-old) and old constant oestrous (19-20-month-old) rats ovariectomized for 7 days were primed with oestradiol-17beta-filled Silastic capsules. Two days later, rats received s.c. progesterone at 09.00 h and then were injected i.p. with either saline or pentobarbital at 13.30 hours. Pentobarbital-treated rats received i.v. pulses of neuropeptide Y, LHRH, a combination of neuropeptide Y and LHRH, or saline, every 30 min from 14.00 to 18.00 h via a jugular cannula. Hourly blood samples were collected between 11.00 and 21.00 h. In old rats, the progesterone induced LH surge was significantly attenuated and delayed as compared to that of young rats. Pentobarbital injection completely blocked the LH surge. Neuropeptide Y pulses alone had no significant effect on LH release. In contrast, LHRH pulses increased LH release in both age groups, although the response was significantly reduced in older rats. While combined pulses of neuropeptide Y and LHRH significantly increased LH release in both young and old rats as compared to that of LHRH alone, the potentiating action of neuropeptide Y on LHRH-induced LH release remained unchanged between the two age groups. These results, together with our recent demonstration of altered hypothalamic neuropeptide Y neuronal activity in middle-aged pro-oestrous rats, suggest that a deficit in neuropeptide Y secretion and action in the hypothalamus, rather than a decrease in pituitary responsiveness to neuropeptide Y, may partially be responsible for the absence of LH surges in old rats. PMID- 10844578 TI - Melanocortin-3 receptor mRNA expression in pro-opiomelanocortin neurones of the rat arcuate nucleus. AB - The melanocortins alpha- and gamma-melanocyte-stimulating hormones (alpha- and gamma-MSH) derive from the pro-opiomelanocortin (POMC) precursor. Melanocortins exert a wide range of biological activities in the brain through activation of at least three distinct melanocortin receptor (MC-R) subtypes. In order to determine whether POMC neurones can modulate their own activity, we looked for the possible expression of the MC3-R gene in POMC-positive cell bodies in the rat hypothalamus. In situ hybridization experiments revealed that the density of MC3 R mRNA is particularly high in the arcuate nucleus which contains the main population of POMC neurones in the brain. The occurrence of MC3-R mRNA in POMC positive cell bodies was demonstrated using a double-labelling in situ hybridization technique. The proportion of POMC neurones expressing MC3-R mRNA was significantly higher in the most rostral (43.5%) than in the most posterior part of the arcuate nucleus (8.2%). These results indicate that melanocortins likely exert a direct regulatory feedback on POMC neurones through activation of MC3-R receptors. Our data also suggest that MC3-R may be involved in the neuroendocrine responses induced by centrally administered melanocortins. PMID- 10844579 TI - The variability of basal action potential firing is positively correlated with bursting in hypothalamic oxytocin neurones. AB - Magnocellular oxytocin neurones are proposed as a suitable system for studying the mechanisms involved in the regulation of neuronal bursting activity. They display high frequency (50 sp./s) bursts of spikes (approximately every 300 s), in response to specific stimuli, which are superimposed on a variable level of basal activity and are tightly co-ordinated as a result of network interactions. The relationship between the strength of the bursting activity (as quantified by burst amplitude and interburst interval) and the characteristics of the interburst basal activity were assessed. During control conditions, mean basal activity and variability of firing increased just before bursts. During experimental conditions leading to burst facilitation, burst amplitude increased and interburst interval decreased while a sustained increase in mean firing rate occurred. Variability of firing (measured by both the standard deviation of firing rate, and the index of dispersion which corrected this standard deviation for differences in mean firing rate), increased demonstrating an increase in spike clustering greater than expected as a result of increased basal activity. When bursting was restrained (i.e. interburst interval increased), mean basal activity increased substantially, but index of dispersion decreased. A narrowing of the interspike interval distribution occurred, indicating increased regularity of firing. The aspect of basal activity most strongly correlated with bursting was variability of firing rate. The strongest correlate of burst amplitude was the standard deviation of mean firing rate, whereas the strongest and most consistent correlate of interburst interval was the index of dispersion. In conclusion, bursting behaviour is most strongly related to the irregularity rather than the level of basal activity. PMID- 10844580 TI - Gonadotropin-releasing hormone exhibits circadian rhythm in phase with arginine vasopressin in co-cultures of the female rat preoptic area and suprachiasmatic nucleus. AB - To determine whether the suprachiasmatic nucleus can drive a circadian release of gonadotropin-releasing hormone (GnRH) in the preoptic area, we measured the release of GnRH, arginine-vasopressin and vasoactive intestinal polypeptide (VIP) in cocultures of the preoptic area and the suprachiasmatic nucleus at 2-h intervals over a period of 120 h. The release of GnRH in cocultures exhibited a significant circadian rhythm in the presence of oestrogen but not in the absence of oestrogen. The period of the GnRH circadian rhythm was the same as that of the arginine-vasopressin circadian rhythm, and different from the VIP circadian rhythm in each coculture. Furthermore, the peak phase of the GnRH rhythm occurred at the time same as that of the arginine-vasopressin rhythm in each coculture. However, the peak phase of the GnRH rhythm was not always the same as that of the VIP rhythm. Administration of arginine-vasopressin significantly increased GnRH release in single preoptic area cultures in the presence of oestrogen, but VIP did not. The result suggests that, in cocultures of the suprachiasmatic nucleus and the preoptic area, arginine-vasopressin neurones drive the circadian release of GnRH in the presence of oestrogen. We suggest that arginine-vasopressin neurones in the suprachiasmatic nucleus mediate the clock information to GnRH neurones in vivo as well. PMID- 10844581 TI - Intracerebroventricular administration of melanin-concentrating hormone suppresses pulsatile luteinizing hormone release in the female rat. AB - Melanin-concentrating hormone (MCH) has been reported to be involved in the regulation of feeding behaviour in rats and mice. Because many neuropeptides that influence ingestive behaviour also regulate reproductive function, the present study was designed to determine if central administration of MCH changes pulsatile secretion of luteinizing hormone (LH) in the rats. Wistar-Imamichi strain female rats were ovariectomized and implanted with oestradiol to produce a moderate inhibitory feedback effect on LH release. The effects of i. c.v. injections of MCH on LH release were examined in freely moving animals. Blood samples were collected every 6 min for 3 h through an indwelling cannula. After 1 h of sampling, MCH (0.1, 1 or 10 microg/animal) or vehicle (saline) was injected into the third cerebroventricle. Because MCH is also reported to affect the hypothalamo-pituitary-adrenal (HPA) axis, which in turn, can influence reproductive function, plasma corticosterone concentrations were determined in the same animals at 30-min intervals during the first and last hours and every 12 min during the second hour of the 3-h sampling period. When expressed as per cent changes, mean plasma LH concentrations after MCH administration were significantly lower in the animals injected with all doses of the peptide compared with vehicle-treated animals; LH pulse frequency was significantly lowered by 1 microg of MCH. Per cent changes in mean plasma corticosterone levels were not significantly affected by MCH administration. These results in oestradiol-treated ovariectomized rats indicate that central MCH is capable of inhibiting pulsatile LH secretion. We have previously shown that 48-h fasting suppresses pulsatile LH release in the presence of oestrogen. Take together, these results raise the possibility that MCH could play a role in mediating the suppression of LH secretion during periods of reduced nutrition. PMID- 10844582 TI - Two olfactory placode derived galanin subpopulations: luteinizing hormone releasing hormone neurones and vomeronasal cells. AB - In adult rodents, the peptide galanin is expressed in a subpopulation of hypothalamic luteinizing hormone-releasing hormone (LHRH) neurones in an activity dependent manner. In this investigation, we examined whether galanin mRNA expression in mice was activated coincident with LHRH mRNA expression, as LHRH neurones differentiate from the olfactory placode. Using in situ hybridization, we show (i) that galanin mRNA is coexpressed in LHRH neurones prenatally, (ii) that there is a decrease in galanin mRNA expression relative to LHRH mRNA expression once LHRH mRNA positive/galanin mRNA positive neurones migrate out of the olfactory pit and into the nasal septum, and (iii) the presence of a novel population of galanin mRNA positive/LHRH mRNA negative expressing neurones in the olfactory pit/vomeronasal organ which do not migrate into the central nervous systenm (CNS). This study demonstrates that there are at least two populations of galanin mRNA expressing neurones arising from the olfactory placode; one that remains in nasal regions, is LHRH mRNA negative and whose function is unknown, and one which is coexpressed with LHRH. In addition, the temporal expression of galanin mRNA in LHRH cells indicates that initial activation and subsequent inactivation of galanin mRNA expression is independent of synaptic CNS connections. PMID- 10844583 TI - The immune-endocrine interaction varies with the duration of the inflammatory process in cardiac surgery patients. AB - The present study investigated the perioperative course of cytokine release and hypothalamic-pituitary-adrenal (HPA) axis activation in relation to the duration of the inflammatory response in cardiac surgery patients. Twelve male patients scheduled for elective coronary artery bypass grafting surgery with cardiopulmonary bypass and general anaesthesia were divided into two study groups: group 1 (n=6) underwent surgery at 13.00 h+/-30 min, group 2 (n=6) at 08.30 h+/-50 min. Blood samples were collected preoperatively and up to the first postoperative day. Postoperatively, on the day of surgery, serum concentrations of the proinflammatory cytokines interleukin (IL)-6, IL-1beta and tumour necrosis factor (TNF)-alpha were not significantly different between the two groups, while blood concentrations of cortisol, adrenocorticotrophic hormone (ACTH) and beta endorphin in group 2 patients were significantly higher than in group 1 patients. Postoperatively, on the day of surgery, ACTH and cortisol concentrations in group 1 patients were positively correlated to the blood concentrations of IL-1beta, IL 6 and TNF-alpha. By contrast, group 2 patients showed no significant relationship between cytokine release and activation of HPA axis at this time. Our results suggest that in patients undergoing cardiac surgery, the cytokine response is initiated before the HPA axis is fully activated. In the early postoperative period, cytokines appear to be involved in the activation of the HPA axis, while in the later postoperative period, high cortisol concentrations may inhibit the release of IL-6. PMID- 10844584 TI - Temperature and photoperiod interact to affect reproduction and GnRH synthesis in male prairie voles. AB - Prairie voles (Microtus ochrogaster), like most rodent species, exhibit a phenotypic polymorphism in reproductive response to winter conditions or to short day lengths in the laboratory. Laboratory studies on seasonally breeding species have traditionally focused on the role of photoperiod in modulating reproduction and other seasonal adaptions. However, because animals use proximate environmental factors in addition to photoperiod to phase seasonal adaptions with the appropriate time of year, the present study investigated the interaction of photoperiod and temperature on reproductive function and the gonadotropin releasing hormone (GnRH) neuronal system. Male prairie voles were housed in either long (LD 16:8) or short (LD 8:16) photoperiods. Voles in each photoperiodic condition were also exposed to either mild (20 degrees C) or low (8 degrees C) temperatures. After 10 weeks, voles were killed and their brains were processed using in situ hybridization for mRNA for proGnRH. The results suggest that GnRH synthesis is not affected by exposure to a single inhibitory proximate factor (i.e. short days or low temperatures alone), even when reproduction is inhibited, whereas a combination of inhibitory proximate factors leads to a decrease in GnRH synthesis (i.e. fewer neurones staining for mRNA for proGnRH). These data suggest that the neuroendocrine mechanisms regulating seasonal alterations in reproductive function are likely to differ between harsh and mild winters. PMID- 10844585 TI - In vitro effects of oestradiol on galanin gene expression in rat anterior pituitary cells. AB - While the pituitary galanin gene is highly responsive to oestrogen stimulation in vivo, in vitro effects of oestrogens on pituitary galanin gene expression have been less studied. We therefore examined the short-term effects of 17beta oestradiol on galanin synthesis by dispersed rat anterior pituitary cell cultures and investigated the mechanisms by which oestrogens may modulate galanin gene expression. 17beta-oestradiol increased galanin mRNA expression in a dose dependent manner, with a maximal increase observed at a concentration of 10-7 M. The 17beta-oestradiol (10-7 M)-induced increase in galanin mRNA expression varied from 3- to 20-fold (average 12-fold) depending upon the experiments and was also time-dependent, reaching significance after 6 h. A 1-h exposure of anterior pituitary cells to 17beta-oestradiol was sufficient to induce markedly galanin mRNA expression after 24 h (by 16-fold) and 48 h (by 25-fold). Tamoxifen administered simultaneously with or following 17beta-oestradiol treatment completely abolished the oestrogen-induced increase of galanin mRNA levels. Cycloheximide (10 microg/ml), a protein synthesis inhibitor, also blocked 17beta oestradiol-induced galanin gene expression. Using transcription blockade by actinomycin D, we observed similar decreases of pituitary galanin mRNA concentrations, in the presence and absence of 17beta-oestradiol, implying no oestrogen effect on mRNA stability. We conclude that oestrogens stimulate rat pituitary galanin gene expression, mainly through a transcriptional mechanism, and that this effect requires persistent binding of the hormone to its nuclear receptor and newly synthesized protein intermediates. PMID- 10844586 TI - The distribution of progesterone receptor immunoreactivity and mRNA in the preoptic area and hypothalamus of the ewe: upregulation of progesterone receptor mRNA in the mediobasal hypothalamus by oestrogen. AB - The distribution of progesterone receptors (PR) was mapped in the hypothalamus of the ewe using immunocytochemistry. These results were confirmed using in situ hybridization with a sheep-specific 35S-labelled riboprobe. In addition, the effect of oestrogen on the level of PR mRNA in the hypothalamus was examined in ovariectomized (OVX) ewes following treatment with an oestrogen implant or without treatment. PR immunoreactive (-ir) cells were readily detected in OVX animals. Labelled cells were observed in four main hypothalamic regions: the preoptic area (POA), including the organum vasculosum of the lamina terminalis, periventricular nucleus (PeVN), ventromedial nucleus (VMN) and the arcuate nucleus (ARC) (including the region ventral to the mamillary recess). In addition, lightly stained PR-ir cells were observed in the supraoptic nucleus and a few PR-ir cells were also found in the diagonal band of Broca. No PR-ir cells were found in the brainstem. PR mRNA-containing cells were found in the same hypothalamic regions as the PR-ir cells. Image analysis of emulsion-dipped slides following in situ hybridization indicated that oestrogen treatment increased (P<0.01) the mean number of silver grains/cell and the density of labelled cells in the VMN and ARC but had no effect on the level of PR mRNA expression in the POA or PeN. The distribution of PR-containing cells in the hypothalamus is similar to that described in other species and all cells were located in nuclei that contain large populations of oestrogen receptor-containing cells. These include regions implicated in the regulation of reproductive neuroendocrine function, and reproductive behaviour. Oestrogen and progesterone synergize to inhibit GnRH secretion and the present results suggest that these functions may involve cells of the VMN and ARC, with oestrogen acting to upregulate PR. PMID- 10844587 TI - Is the mineralocorticoid receptor in Brown Norway rats constitutively active? AB - In a previous study using corticosterone treatment of adrenalectomized rats, we hypothesized that mineralocorticoid receptor (MR)-related mechanisms are constitutively active and that glucocorticoid receptor (GR)-mediated mechanisms are more efficient in Brown Norway rats compared to Fischer 344 (F344) rats. In order to discriminate the mineralocorticoid from the glucocorticoid actions exerted by corticosterone, F344 and Brown Norway adrenalectomized rats were treated with increasing doses (1, 5 and 25 microg/ml of drinking water) of deoxycorticosterone (DOC, MR-specific ligand) or RU 28362 (GR-specific ligand). These rats were compared with long-term adrenalectomized (ADX) untreated rats and sham-ADX rats. This study confirms our previous results, notably the lack of effect of ADX on body weight and fluid intake in Brown Norway rats. Moreover, DOC treatment had no effect in Brown Norway rats whereas the higher dose restored fluid intake of the F344 ADX group to sham values. These results support the hypothesis of a constitutive activation of the MR and therefore the insensitivity of this receptor to its ligand in Brown Norway rats. Alternatively, RU 28362 treatment induced greater weight loss, decrease in food intake, anxiolysis, thymus involution, and decrease in plasma transcortin concentration and pituitary corticosteroid receptor densities in Brown Norway rats than in F344 rats, which is consistent with greater efficiency of GR mechanisms in Brown Norway rats than in F344 rats. Therefore, these strains are of great utility to disentangle MR and GR effects on complex phenotypes. PMID- 10844588 TI - Trafficking of the vasopressin and oxytocin prohormone through the regulated secretory pathway. AB - The trafficking of prohormones and of regulated secretory proteins in general has been studied extensively in the last decades of the last century. Prohormone trafficking starts with correct folding and subsequently efficient sorting into the secretory granule of the regulated secretory pathway. The vasopressin/oxytocin prohormone is particularly interesting for studying protein trafficking, because the physicochemical properties and three-dimensional structure have been largely elucidated. In the case of pro-vasopressin and pro oxytocin, folding and sorting depend completely on both intramolecular and intermolecular interactions. Proper folding is guided by the hormone-neurophysin association and the sorting event relies on the aggregative properties of the neurophysin domain in the prohormone, as well as a specific sorting signal, which is revealed when the aggregative property of the neurophysin domain is deleted. A comprehensive mechanism for trafficking of the vasopressin/oxytocin prohormone from the endoplasmic reticulum to the secretory granule is proposed. PMID- 10844589 TI - Antiproteinase 3- and antimyeloperoxidase-associated vasculitis. AB - Antiproteinase 3- and antimyeloperoxidase-associated vasculitis. Wegener's granulomatosis, microscopic polyangiitis, and idiopathic pauci-immune necrotizing crescentic glomerulonephritis (NCGN) are strongly associated with antineutrophil cytoplasmic autoantibodies (ANCAs) directed against either proteinase 3 (anti PR3) or myeloperoxidase (anti-MPO). This has led some investigators to prefer combining these diseases under the common heading of ANCA-associated vasculitides. However, it is increasingly recognized that there are characteristic differences between patients with anti-PR3 and those with anti-MPO associated vasculitis. This review focuses on the clinical, histopathologic, and possibly pathophysiologic differences between anti-PR3- and anti-MPO-associated vasculitis. Although there is considerable overlap, the anti-PR3- and anti-MPO associated vasculitides are each characterized by particular clinical and histopathological findings. Extrarenal organ manifestations and respiratory tract granulomas occur more frequently in patients with anti-PR3 than in those with anti-MPO. Anti-PR3-positive patients with NCGN generally have a more dramatic deterioration of their renal function compared with anti-MPO-positive patients. The term "ANCA-associated vasculitis" is considered as a useful concept in the presence of systemic vasculitis. Likewise, in the presence of vasculitis, the terms "anti-PR3-associated vasculitis" and "anti-MPO-associated vasculitis" are useful concepts. PMID- 10844590 TI - The family of bone morphogenetic proteins. AB - The family of bone morphogenetic proteins. Bone morphogenetic proteins (BMPs) are secreted signaling molecules belonging to the transforming growth factor-beta (TGF-beta) superfamily of growth factors. The first BMPs were originally identified by their ability to induce ectopic bone formation when implanted under the skin of rodents. In this ectopic overexpression assay, there was a recapitulation of all the events occurring during skeletogenesis. This latter aspect indicated that these molecules could play important roles during development. More than 30 BMPs have been identified to date. The study of their expression pattern as well as the analysis of spontaneously mutated or genetically depleted mice have demonstrated a much broader range of function. These activities are mainly localized at sites of epithelial-mesenchymal interactions, including but not restricted to the skeleton. This review presents our current knowledge about the functions of BMPs during skeleton development as well as in many other biologic processes. PMID- 10844591 TI - XDH gene mutation is the underlying cause of classical xanthinuria: a second report. AB - BACKGROUND: Classical xanthinuria is a rare autosomal recessive disorder characterized by excessive excretion of xanthine in urine. Type I disease results from the isolated deficiency of xanthine dehydrogenase (XDH), and type II results from dual deficiency of XDH and aldehyde oxidase. The XDH gene has been cloned and localized to chromosome 2p22-23. The aim of this study was to characterize the molecular basis of classical xanthinuria in an Iranian-Jewish family. METHODS: The apparently unrelated parents originated from a community in which consanguineous marriages are common. Subtyping xanthinuria was attempted by homozygosity mapping using microsatellite markers D2S352, D2S367, and D2S2374 in the vicinity of the XDH gene. Mutation detection was accomplished by PCR-SSCP screening of all 36 exons and exon-intron junctions of the XDH gene, followed by direct sequencing and confirmation of sequence alteration by restriction analysis. RESULTS: The index case was homozygous for all three microsatellite markers analyzed. The expected frequency of this genotype in a control population was 0. 0002. These results suggested that xanthinuria in the patient is linked to the XDH gene. Consequently, a 1658insC mutation in exon 16 of the XDH gene was identified. The 1658insC mutation was not detected in 65 control DNA samples. CONCLUSION: A molecular approach to the diagnosis of classical xanthinuria type I in a female patient with profound hypouricemia is described. Linkage of xanthinuria to the XDH locus was demonstrated by homozygosity mapping, and a 1658insC mutation, predicting a truncated inactive XDH protein, was identified. These results reinforce the notion that mutations in the XDH gene are the underlying cause of classical xanthinuria type I. PMID- 10844592 TI - Identification of novel genes expressed during metanephric induction through single-cell library screening. AB - BACKGROUND: Development of the mature kidney is dependent on a series of inductive events between a portion of the epithelial bud at the distal end of the nephric duct and a neighboring domain of committed metanephric mesenchyme. Several genes have been identified to date that are critical in the inductive process. For example, the deletion of Bmp7 from the mouse genome results in dysgenesis or agenesis of the kidney. These findings suggest that Bmp7 controls the expression of genes important for nephrogenesis, but the identity of these genes has remained largely undetermined. METHODS: Single cells were isolated from mouse metanephric mesenchyme during the time of induction (between E11.0 and E11.5) and cDNA libraries constructed from induced and uninduced tissue. Subtractive hybridization was performed to isolate genes that were expressed during E11.5 but not E11.0. RESULTS: Using this approach, we identified eight previously known genes, three of which were known to be involved in metanephric induction, thus validating our approach, and nine novel genes. Eight of these genes were completely novel, whereas one was similar to a member of the yeast Anaphase Promoting Complex. CONCLUSIONS: Through subtractive hybridization of mouse E11.0 and E11.5 metanephric mesenchyme single-cell cDNA libraries, we have identified novel genes that are candidates for involvement in nephrogenesis through their up-regulation during the inductive process. PMID- 10844593 TI - Cox-2 and osteopontin in cocultured platelets and mesangial cells: role of glucocorticoids. AB - BACKGROUND: Glomerular inflammation is characterized by a consecutive infiltration of immunoreactive cells. To mimic the early phase of glomerular injury, a coculture system of platelets and rat renal mesangial cells was established. As prototypes, the inflammation-related proteins cyclooxygenase-2 (Cox-2) and the chemotactic protein osteopontin (OPN) were investigated. METHODS: The expression of OPN and Cox-2 mRNA and protein was determined by Northern and Western blot analyses. RESULTS: Coincubation of platelets and mesangial cells led to a rapid, transient induction of Cox-2 mRNA, which peaked at two hours, whereas OPN and monocyte chemoattractant protein-1 (MCP-1) were induced at later time points. The induction of Cox-2 mRNA was concentration dependent and highly reproducible when platelets of different donors were investigated. Partial Cox-2 induction was observed when supernatants of preactivated platelets were incubated with mesangial cells. The inhibition of the signaling pathways of platelet derived growth factor (PDGF) and epidermal growth factor (EGF) or interference with Gi-protein signaling partially inhibited platelet-induced Cox-2 expression. Down-regulation of protein kinase C (PKC), which is a common signaling module in many pathways leading to Cox-2 induction, almost completely abrogated platelet induced Cox-2 expression. The time pattern of Cox-2 and OPN expression suggested that Cox-2 might play a role in OPN induction. The up-regulation of OPN was dependent on de novo protein synthesis and was induced by high levels of exogenous prostaglandin E2 (PGE2; 10 micromol/L). Endogenous PGE2, however, proved not to be essential for OPN mRNA expression, because inhibition of Cox activity did not change OPN mRNA levels. Dexamethasone inhibited Cox-2 mRNA induction but increased OPN mRNA and protein expression. CONCLUSION: These data indicate that Cox-2 and OPN are independently up-regulated upon interaction of platelets and mesangial cells. PMID- 10844594 TI - alpha-MSH inhibits induction of C/EBPbeta-DNA binding activity and NOS2 gene transcription in macrophages. AB - BACKGROUND: alpha-Melanocyte-stimulating hormone (alpha-MSH) is an endogenous tridecapeptide that exerts anti-inflammatory actions and abrogates postischemic renal injury in rodents. alpha-MSH inhibits lipopolysaccharide (LPS)-induced gene expression of several cytokines, chemokines, and nitric oxide synthase-2 (NOS2), but the molecular mechanisms underlying these effects have not been clearly defined. To test the hypothesis that alpha-MSH inhibits the expression of inducible trans-activating factors involved in NOS2 regulation, we used RAW 264.7 macrophage cells to examine the effects of alpha-MSH on the activation of nuclear factor-kappaB (NF-kappaB) and CCAAT/enhancer binding protein-beta (C/EBPbeta), trans-acting factors known to be involved in LPS + interferon (IFN)-gamma induction of the NOS2 gene. METHODS: Gel shift assays were performed to identify NF-kappaB and C/EBP DNA binding activities in LPS + IFN-gamma-treated RAW 264.7 cells in the presence and absence of alpha-MSH. NOS2 promoter assays were conducted to identify the effects of alpha-MSH on LPS + IFN-gamma-mediated induction of NOS2 transcription. RESULTS: Gel shift assays demonstrated LPS + IFN gamma induction of NF-kappaB and C/EBP family protein-DNA complexes in nuclei harvested from the cells. Supershift assays revealed that the C/EBP complexes were comprised of C/EBPbeta, but not C/EBPalpha, C/EBPdelta, or C/EBPepsilon. alpha-MSH (100 nmol/L) inhibited the LPS + IFN-gamma-mediated induction of nuclear DNA binding activity of C/EBPbeta, but not that of NF-kappaB (in contrast to reports in other cell types), as well as the activity of a murine NOS2 promoter-luciferase construct. In contrast, alpha-MSH (100 nmol/L) had no effect on the induction of NOS2 promoter-luciferase genes harboring deletion or mutation of the C/EBP box. CONCLUSIONS: These data indicate that alpha-MSH inhibits the induction of C/EBPbeta DNA binding activity and that this effect is a major mechanism by which alpha-MSH inhibits the transcription of the NOS2 gene. The inability of alpha-MSH to inhibit LPS + IFN-gamma induction of NF-kappaB in murine macrophage cells, which contrasts with inhibitory effects of the neuropeptide in other cell types, suggests that cell-type-specific mechanisms are involved. PMID- 10844595 TI - Activated STAT1 suppresses proliferation of cultured rat mesangial cells. AB - BACKGROUND: JAK-STAT signaling has been shown to promote development and proliferation in lymphopoietic and hematopoietic lineages. We investigated the effect of activated STAT1 on mesangial cell proliferation. METHODS: Rat mesangial cells of primary culture (rMCs) were used in the following experiments: (1) Whole cell lysates were immunoblotted against JAK1 and JAK2. (2) Whole cell lysates and nuclear proteins were extracted from rMCs with or without treatment with interferon-gamma, and immunoblotting was performed against both STAT1 and tyrosine (701)-phosphorylated STAT1. (3) rMCs and rMCs electroporated with either wild-type STAT1, mutated STAT1, or antibody against STAT1 were incubated with interferon-gamma for 20 hours, followed by a further incubation with [3H] thymidine for four hours. RESULTS: JAK1, JAK2, and STAT1 were detected in whole cell lysates, suggesting that JAK-STAT signaling could be activated by interferon gamma (INF-gamma). Using an antibody specific for tyrosine-phosphorylated STAT1, we detected signal in the INF-gamma-treated nuclear extracts, which showed translocation of phosphorylated STAT1 to the nucleus. [3H]-thymidine incorporation in the presence of INF-gamma was significantly lower than that of control in a dose-dependent manner. The introduction of wild-type STAT1 enhanced the effect of interferon-gamma and decreased [3H]-thymidine incorporation, whereas tyrosine-mutated (Y701F) STAT1 and SH2 domain (R602T)-mutated STAT1 reversed INF-gamma-induced suppression of [3H]-thymidine incorporation. Electroinjected antibody against STAT1 increased [3H]-thymidine incorporation upon stimulation with INF-gamma. CONCLUSION: STAT1 activated by interferon-gamma suppresses mesangial cell proliferation. PMID- 10844596 TI - Effect of metabolic acidosis on the growth hormone/IGF-I endocrine axis in skeletal growth centers. AB - BACKGROUND: Chronic metabolic acidosis (CMA) adversely affects bone metabolism and skeletal growth. Given the cardinal role played by the local growth hormone (GH)/insulin-like growth factor-I (IGF-I) in promoting cell proliferation and differentiation in growth plates, we tested the effect of CMA on the GH/IGF-I axis in a skeletal growth center. METHODS: We employed an in vitro organ culture system using the murine mandibular condyle as a model for endochondral active growth center. Condyles from six-day-old ICR mice were cultured in BGJb medium of either neutral pH (pH approximately 7.4) or acidic pH (pH approximately 7.15). After 24, 48, 72, and 96 hours of culture, the condyles were washed, fixed in formaldehyde, and processed for paraffin embedding. We assessed histologic markers of the growth center. In addition, the protein level and mRNA expression for the different components of the GH/IGF-I axis were evaluated by immunohistochemistry and in situ hybridization, respectively. Finally, we evaluated the effect of acidosis on the biological functions mediated by GH and IGF-I (namely, proliferation and differentiation of cartilage cells in the active growth center). RESULTS: Following three to four days in acidic conditions, there was a marked reduction in the size of young chondrocytic population, suggesting a defect in the process of endochondral differentiation. Immunohistochemistry and in situ hybridization analyses revealed a marked reduction in the expression of the IGF-I receptor, as well as in the GH receptor. These changes were already evident after 48 hours of incubation in acidic conditions. At 48 hours of acidosis, there was also a marked reduction in the expression of IGF-I both under basal conditions (nonstimulated) and following stimulation with GH. The expression of IGF binding protein 2 (IGFBP-2) and IGFBP-4, which serve as negative modulators of IGF-I, was enhanced in CMA. IGF-I markedly stimulated chondrocytic proliferation (assessed by BrdU incorporation into DNA) and differentiation (assessed as cartilage specific proteoglycan expression). These responses were markedly attenuated in acidic conditions. CONCLUSION: CMA exerts an anti-anabolic effect in bone growth centers, which is partly related to a state of resistance to GH and IGF-I, created by CMA. This phenomenon may underlie the disturbance in longitudinal bone growth in CMA (that is, renal tubular acidosis) and may contribute to renal osteodystrophy in patients suffering from chronic renal failure. PMID- 10844597 TI - Estrogen effects on triglyceride metabolism in analbuminemic rats. AB - BACKGROUND: Triglyceride (TG) levels are normally lower in female rats, while the opposite is the case in the Nagase analbuminemic rats (NAR). Increased TG levels in normal males are caused by a testosterone-mediated decrease in postheparin (PH) lipoprotein lipase (LpL). Castration of males reduces TG, while castration of females is without effect. TG levels are reduced by castration of the female NAR, suggesting that estrogen rather than testosterone causes hypertriglyceridemia in this strain. The mechanism for this increase is unknown. METHODS: We measured secretion of very-low density lipoprotein (VLDL) TG using Triton WR 1339 clearance as the disappearance from blood of 3H-trioleate and 14C cholesterol-labeled chylomicrons (CM), and the activity of the PH lipases: LpL and hepatic lipase (HL). All were determined in Sprague-Dawley (SD) and NAR female, male, and ovariectomized (OVX) rats. RESULTS: TG levels were significantly greater in female NAR in comparison to all other groups. Ovariectomy of NAR significantly ameliorated hypertriglyceridemia. VLDL TG secretion was significantly greater in intact female NAR compared with all other groups. There were no other differences in VLDL TG secretion among the other groups. The clearance of CM was greatest in female SD rats, and OVX had no effect. NAR cleared CM less well than did SD rats (P < 0.001), but among NAR, clearance was greatest in OVX NAR and male NAR (P < 0. 002). Both PH LpL activity and HL activity were lowest in female NAR (P < 0.05). Ovariectomy partially corrected the defect in HL (P < 0. 05). CONCLUSION: TG levels in female NAR are in part a result of increased VLDL-TG secretion, an effect mediated by estrogen. The presence of an estrogen-mediated catabolic defect that was alleviated by OVX was also observed. This catabolic defect is likely a result of an estrogen mediated decrease both in LpL and HL expressed only in the presence of analbuminemia. PMID- 10844598 TI - L-Arginine treatment may prevent tubulointerstitial nephropathy caused by germanium dioxide. AB - BACKGROUND: Long-term oral ingestion of germanium dioxide (GeO2) causes progressive renal failure derived from tubulointerstitial nephropathy in humans and animals. The characteristic of GeO2-induced nephropathy is the renal tissue injury persisting for a long time, even after cessation of GeO2 ingestion. However, a treatment that can suppress the long-lasting renal tissue injury has not yet been established. METHODS: Using the methods of immunohistochemistry and reverse transcription-polymerase chain reaction, we examined the expression of ED1-positive cells (macrophages/monocytes), transforming growth factor (TGF) beta1 mRNA and protein and collagen type IV mRNA and protein in the kidneys of rats with GeO2-induced nephropathy. Concomitantly, the effects of L-arginine treatment on their expression was explored in the kidneys of rats with GeO2 induced nephropathy. RESULTS: Chronic administration of GeO2 caused tubulointerstitial nephropathy characterized by leukocyte invasion into the enlarged tubulointerstitial space in rats. The expression of ED1-positive cells, TGF-beta1 protein and collagen type IV protein was markedly increased in the tubulointerstitium of the renal cortex from rats with GeO2-induced nephropathy. Similarly, TGF-beta1 and collagen type IV mRNA were significantly enhanced in the renal cortex of rats with GeO2-induced nephropathy. A small number of tubulointerstitial cells expressing TGF-beta1 protein were also observed in the renal cortex of rats with GeO2-induced nephropathy. However, L-arginine treatment led to a parallel decrease in the expression of ED1-positive cells, TGF-beta1 mRNA and collagen type IV mRNA and protein in rats with GeO2-induced nephropathy. CONCLUSIONS: In general, collagen synthesis is driven by TGF-beta1 in the fibrotic process associated with a variety of renal disorders. TGF-beta1 is secreted by TGF-beta1 producing cells such as macrophages, fibroblasts and myofibroblasts. Thus, the present study indicates that the expression of collagen type IV may be mediated by TGF-beta1 released from invading macrophages and, to a lesser extent, released from tubulointerstitial cells, presumably fibroblasts and/or myofibroblasts in GeO2-induced nephropathy. L-Arginine treatment inhibits collagen type IV synthesis possibly by suppressing macrophage invasion and the resultant TGF-beta1 expression in this nephropathy. L-Arginine treatment may be beneficial in the prevention of tubulointerstitial fibrosis, which is considered to be the terminal stage of GeO2-induced nephropathy. PMID- 10844599 TI - Angiotensin III increases MCP-1 and activates NF-kappaB and AP-1 in cultured mesangial and mononuclear cells. AB - BACKGROUND: Monocyte infiltration is a common feature of renal diseases. Angiotensin II (Ang II) participates in inflammatory cell infiltration in the kidney. However, the influence of other peptides of the renin-angiotensin system, such as the N-terminal Ang II degradation product Ang III, has not been addressed. METHODS: In cultured renal and mononuclear cells, we investigated whether Ang III is involved in monocyte recruitment through the regulation of the chemokine, monocyte chemoattractant protein-1 (MCP-1; Northern blot, Western blot, immunofluorescence, and chemotaxis), and the activation of transcription factors, nuclear factor kappaB (NF-kappaB) and activating protein-1 (AP-1; electrophoretic mobility shift assay). RESULTS: In cultured rat mesangial and mononuclear cells, Ang III increased MCP-1 gene expression and protein levels. Supernatants from Ang III-treated mesangial cells showed increased chemoattractant activity for monocytes, which was partially inhibited by the addition of anti-MCP-1 antibody. Ang III elicited a rapid NF-kappaB activation (8 fold, after 30 min), showing a kinetics and intensity similar to that observed with Ang II and tumor necrosis factor-alpha. The maximal NF-kappaB activation was correlated with nuclear translocation of p50 and p65 subunits and disappearance of cytosolic IkappaB. Ang III also activated AP-1 (5-fold, after 18 h), while SP 1 was unchanged. Two NF-kappaB inhibitors abolished the Ang III-induced MCP-1 mRNA expression, suggesting that overexpression of this chemokine is mediated, at least in part, by NF-kappaB activation. CONCLUSIONS: Ang III activates the transcription factors NF-kappaB and AP-1 and increases the expression of related genes, such as MCP-1. Our study describes a novel and potent proinflammatory action of this Ang degradation product, expanding the present view of the renin angiotensin system. PMID- 10844600 TI - Glomerulosclerosis in mice transgenic for human insulin-like growth factor binding protein-1. AB - BACKGROUND: The growth hormone (GH)/insulin-like growth factor (IGF) system is thought to participate in the glomerulosclerosis process. Because IGF-binding proteins (IGFBPs) modulate IGF actions and hence GH secretion, this study assessed whether mice transgenic for human IGFBP-1 have altered susceptibility to glomerulosclerosis. METHODS: A line of transgenic mice that express human IGFBP-1 mRNA in the liver under the control of the alpha1-antitrypsin promoter has been obtained, and morphological changes in the kidney tissue were assessed. Glomerulosclerosis was identified using light microscopy, light microscopic morphometry, and electron microscopy. Extracellular matrix components were analyzed by immunohistochemistry. RESULTS: There was a marked increase in mesangial extracellular matrix area in homozygous transgenic mice at three months of age as compared with heterozygous transgenic mice and nontransgenic littermates. These changes were not associated with alterations in glomerular volume or cellularity. The expansion of extracellular matrix area was related to a marked increase in laminin and type IV collagen and to the appearance of type I collagen. CONCLUSIONS: These observations indicate that the enhanced expression of IGFBP-1 may result in the development of glomerulosclerosis without glomerular hypertrophy. The changes are potentially related to a decrease in IGF-I availability and/or to an IGF-I-independent role of IGFBP-1. PMID- 10844601 TI - Thrombin is a novel regulator of hexokinase activity in mesangial cells. AB - BACKGROUND: Hexokinase (HK) activity is fundamentally important to cellular glucose uptake and metabolism. Phorbol esters increase both HK activity and glucose utilization in cultured mesangial cells via a protein kinase C (PKC)- and extracellular signal-regulated kinases 1 and 2 (ERK1/2)-dependent mechanism. In adult kidneys, increased HK activity has been reported in both glomerular injury and in diabetes, but the mechanisms responsible for these changes are unknown. Thrombin, a known activator of both PKC and ERK1/2, is increased in the settings of renal injury and diabetes. Thus, thrombin may contribute to the observed changes in HK activity in vivo. METHODS: Thrombin and thrombin receptor agonists were tested for the ability to increase HK activity and glucose metabolism in murine mesangial (SV40 MES 13) cells. ERK1/2 activation was also evaluated in parallel. Thrombin inhibition (hirudins), PKC depletion, Ser-Thr kinase inhibition (H-7), MEK1/2 inhibition (PD98059), pertussis toxin (PTX), and general inhibitors of transcription or translation were then tested for the ability to attenuate these effects. RESULTS: Thrombin (>/=0.01 U/mL) mimicked the effect of phorbol esters, increasing HK activity> 50% within 12 to 24 hours (P < 0.05). This effect was inhibited by hirudins, mimicked by thrombin receptor agonists, and accompanied by increased Glc utilization. H-7, PD98059, and general inhibitors of transcription or translation-but not PTX-prevented thrombin-induced HK activity at 24 hours. PKC depletion and PD98059 also blocked the associated phosphorylation and activation of ERK1/2. CONCLUSIONS: Thrombin increases mesangial cell HK activity via a PTX-insensitive mechanism involving thrombin receptor activation, PKC-dependent activation of ERK1/2, and both ongoing gene transcription and de novo protein synthesis. As such, thrombin is a novel regulator of HK activity in mesangial cells and may play a role in coupling renal injury to metabolism. PMID- 10844602 TI - Glucocorticoid modulates Na+/H+ exchange activity in vascular smooth muscle cells by nongenomic and genomic mechanisms. AB - BACKGROUND: In vascular smooth muscle cells (VSMCs), Na+/H+ exchange (NHE) plays an important role in intracellular pH (pHi) regulation. The genomic effect of glucocorticoid (GC) on NHE activity has been suggested in VSMCs. However, the nongenomic and genomic effects of GC on NHE activity and the underlying intracellular signaling mechanisms have not yet been demonstrated in VSMCs. Also, it is not known whether there are specific surface-binding sites of GC to the plasma membrane of VSMCs. METHODS: The effects of short (3 h)- and long (24 h) term exposure to corticosterone (CORTI) on NHE activity were studied in cultured rat aortic VSMCs by using pHi measurement with the pH-sensitive fluorescent dye 2'7'-bis(carboxyethyl)-5(6)-carboxyfluorescein. The NHE activity was calculated from the initial rate of Na+-dependent pHi recovery after the acid load. RESULTS: Short-term exposure of VSMCs to CORTI (10-6 mol/L) increased NHE activity, whereas long-term exposure to CORTI decreased it. The inhibitors of gene transcription (actinomycin D) and of protein synthesis (cycloheximide) did not affect the short-term effect of CORTI on NHE activity, but inhibited the long term effect of CORTI on NHE activity. The cytosolic GC receptor (GR) antagonist (RU38486) inhibited both the short- and long-term effects of CORTI on NHE activity, but the cytosolic mineralocorticoid receptor antagonist (spironolactone) did not influence either the short- or long-term CORTI effects. Two protein kinase C (PKC) inhibitors (staurosporine A and calphostin C) and PKC down-regulation [24-h pre-exposure to phorbol 12-myristate 13-acetate (PMA)] inhibited both short- and long-term CORTI effects. Exposure to PMA for three hours mimicked the short-term CORTI effect. The short-term CORTI effect was inhibited by the disruptor of microtubule (colchicine), but not by the disruptor of filamentous-actin (cytochalasin B). The long-term exposure to CORTI decreased NHE (NHE-1) mRNA levels to 0.65 times the control level, whereas the short-term exposure to CORTI caused no effect. Scatchard analysis of [3H]CORTI surface binding to VSMCs showed a single class of CORTI binding sites with a Bmax of 876.2 fmol per mg of cell protein and a Kd of 12.2 nmol/L. RU38486 also inhibited [3H]CORTI surface binding to VSMCs. CONCLUSIONS: In VSMCs, NHE activity is stimulated by short-term exposure to CORTI, but is inhibited by long-term exposure to CORTI. The short-term stimulatory effect of CORTI on NHE activity is independent of gene transcription and protein synthesis, is mediated through the CORTI surface receptor, and occurs through a microtubule-dependent process. The long-term inhibitory effect of CORTI on NHE activity requires gene transcription and protein synthesis and occurs only through the cytosolic GR. The short- and long-term effects of CORTI on NHE activity occur via PKC activation. Therefore, CORTI differentially modulates NHE activity in VSMCs by nongenomic and genomic mechanisms. PMID- 10844603 TI - A selective cyclooxygenase-2 inhibitor decreases proteinuria and retards progressive renal injury in rats. AB - BACKGROUND: We have previously shown that cyclooxygenase-2 (COX-2) expression is low in the renal cortex of adult rats, but is increased in macula densa/cortical thick ascending limb and in glomerular podocytes after subtotal renal ablation. METHODS: To evaluate the functional consequences of this increased COX-2 expression, male rats were subjected to subtotal renal ablation and divided into four groups: (1) treatment with the selective COX-2 inhibitor SC58236, (2) treatment with vehicle, (3) treatment with the angiotensin-converting enzyme inhibitor enalapril, and (4) treatment with enalapril + SC58236. The administration of drugs was begun on the third day after ablation and continued for 6 to 10 weeks. RESULTS: Within one week after ablation, vehicle-treated rats developed hypertension. Although enalapril led to significant reductions in blood pressure, either alone or in combination with the COX-2 inhibitor, SC58236 alone did not significantly alter ablation-induced hypertension. However, the SC58236 treated animals exhibited levels of proteinuria at six weeks after ablation that were comparable to those seen with enalapril (vehicle, 47 +/- 4; enalapril, 27 +/ 2; SC58236, 30 +/- 2 mg/day; N = 7, P < 0.01, each group compared with vehicle), and continued SC58236 treatment led to persistent reductions in proteinuria at 10 weeks after renal ablation (vehicle, 77 +/- 4; SC58236, 50 +/- 4 mg/day; N = 6, P < 0. 01). SC58236 treatment also significantly reduced the percentage of glomeruli exhibiting segmental or global sclerosis at 10 weeks (32.6 +/- 7.8% vs. 10.9 +/- 2.8%, N = 6, P < 0.03). Furthermore, SC58236 treatment partially inhibited increases in transforming growth factor-beta1 mRNA expression and increases in collagen III and collagen IV mRNA expression. CONCLUSIONS: These studies indicate that chronic treatment with a specific COX-2 inhibitor may retard the progression of progressive renal injury, and suggest that such compounds can be used in combination with angiotensin-converting enzyme inhibitors. Further studies are required to determine the mechanism by which COX 2 inhibition is renoprotective. PMID- 10844604 TI - Decreased urinary excretion of vascular endothelial growth factor in idiopathic membranous glomerulonephritis. AB - BACKGROUND: Membranous glomerulonephritis (MGN) has, for unknown reasons, an unpredictable and highly variable clinical course. Vascular endothelial growth factor (VEGF) enhances endothelial cell proliferation, angiogenesis, microvascular permeability, and monocyte chemotaxis, and it activates proteinases. In normal kidneys, it is predominantly expressed by glomerular podocytes, where its physiological function and role in development of renal diseases are obscure. This study was designed to evaluate the urinary excretion of VEGF in MGN compared with several other glomerular disease and to asses its relationships to the clinical activity of MGN. METHODS: Urinary VEGF was studied during renal biopsy using a sandwich enzyme immunoassay from 30 patients with idiopathic MGN, 8 with minimal change glomerulonephritis, 10 with focal segmental glomerulosclerosis (FSGS), 8 with necrotizing glomerulonephritis associated with systemic vasculitis, and 12 with diabetic nephropathy. In addition, 33 healthy controls were examined. Fifteen patients with MGN were re-evaluated 12 months later, and the evolution of proteinuria was compared with changes in urinary VEGF excretion. RESULTS: In healthy control subjects, urinary VEGF excretion was 68 +/ 10 (95% CI, 49 to 88) ng/mmol creatinine (UCr). In MGN, the excretion was decreased to 16 +/- 3 (CI, 10 to 23) ng/mmol crea (P < 0.0001, ANOVA), whereas in minimal change glomerulonephritis and diabetic nephropathy, it was unchanged [55 +/- 14 (CI, 24 to 86) and 101 +/- 25 (CI, 45 to 156) ng/mmol UCr, respectively, P = NS]. In vasculitis and FSGS patients, the excretion was higher than normal [184 +/- 68 (CI, 24 to 344), P = 0.01, and 160 +/- 29 (CI 95 to 226), P = 0.002 ng/mmol UCr, respectively]. The excretion did not correlate with serum VEGF, renal function, or proteinuria. In the follow-up of 15 patients, improving MGN (decreasing proteinuria) was associated with increasing VEGF excretion, while persistent disease (no change or increase of proteinuria) was associated with constantly low urinary VEGF excretion. The change in urinary protein excretion over one year correlated inversely with the change in urinary VEGF (r = -0.57, P = 0.026). CONCLUSIONS: MGN is associated with decreased urinary VEGF compared with normal subjects, which is in contrast with other proteinuric diseases. Moreover, decreasing clinical activity (proteinuria) is accompanied by increasing VEGF excretion. Urinary VEGF may serve as an indicator of activity of MGN. PMID- 10844605 TI - Cytotoxic effect of Shiga toxin-1 on human glomerular epithelial cells. AB - BACKGROUND: Shiga toxin-1 (Stx-1) has been implicated in the pathogenesis of postdiarrheal hemolytic-uremic syndrome (Stx HUS). Endothelial cells had been felt to be the primary renal target of Stx-1; however, recent studies suggest that renal epithelial cells may also be responsive. To further examine this issue, we evaluated the responsiveness of human glomerular epithelial cells (GECs) to the cytotoxic effects of Stx-1. METHODS: Cultured GECs were exposed to Stx-1 in the presence and absence of a variety of inflammatory factors likely to be elevated in the kidney or serum of patients with Stx HUS. Cell survival, protein synthesis, total cell Gb3 levels and synthesis, and Stx-1 binding were measured. RESULTS: GECs were sensitive to Stx-1, with an LD50 of approximately 10 7 g/L (1.4 pmol/L). Interleukin-1 (IL-1), lipopolysaccharide (LPS), tumor necrosis factor-alpha (TNF-alpha), and butyrate increased Stx-1 cytotoxicity and total cell Gb3 levels. These agents, with the exception of TNF-alpha, also increased Stx-1 binding to GECs. IL-6 failed to alter Stx-1 toxicity, binding, or Gb3 content. CONCLUSIONS: These studies indicate that GECs are sensitive to the cytotoxic effects of Stx-1 and that inflammatory factors can increase toxin responsiveness. GECs may be a target of Stx-1 action in Stx HUS. PMID- 10844606 TI - Endotoxin and nanobacteria in polycystic kidney disease. AB - BACKGROUND: Microbes have been suspected as provocateurs of polycystic kidney disease (PKD), but attempts to isolate viable organisms have failed. Bacterial endotoxin is the most often reported microbial product found in PKD fluids. We assessed potential microbial origins of endotoxin in cyst fluids from 13 PKD patients and urines of PKD and control individuals. METHODS: Fluids were probed for endotoxin and nanobacteria, a new bacterium, by the differential Limulus Amebocyte Lysate assay (dLAL), genus-specific antilipopolysaccharide (LPS) antibodies, monoclonal antibodies to nanobacteria, and hyperimmune serum to Bartonella henselae (HS-Bh). Selected specimens were also assessed by transmission electron microscopy (TEM) and nanobacterial culture methods. RESULTS: LPS or its antigenic metabolites were found in more than 75% of cyst fluids tested. Nanobacteria were cultured from 11 of 13 PKD kidneys, visualized in 8 of 8 kidneys by TEM, and immunodetected in all 13 PKD kidneys. By immunodetection, nanobacterial antigens were found in urine from 7 of 7 PKD males, 1 of 7 PKD females, 3 of 10 normal males, and 1 of 10 normal females. "Nanobacterium sanguineum" was dLAL positive and cross-reactive with antichlamydial LPS and HS-Bh. Some cyst fluids were also positive for LPS antigens from Escherichia coli, Bacteroides fragilis and/or Chlamydia, and HS-Bh, as were liver cyst fluids from one patient. Tetracycline and citrate inhibited nanobacterial growth in vitro. CONCLUSION: Nanobacteria or its antigens were present in PKD kidney, liver, and urine. The identification of candidate microbial pathogens is the first step in ascertaining their contribution, if any, to human disease. PMID- 10844607 TI - Ischemic acute renal failure: long-term histology of cell and matrix changes in the rat. AB - BACKGROUND: The cellular infiltration and matrix accumulation accompanying acute renal ischemia and reperfusion have been frequently noted but poorly defined. The long-term consequences of ischemia may irreversibly damage the kidney. METHODS: Female Sprague-Dawley rats (200 g) underwent unilateral nephrectomy. After five days, the left renal pedicle was occluded for 45 minutes. Animals were sacrificed at 0, 1, 2, 4, 8, 16, 32, 64, and 180 days postischemia (N = 6). Immunohistochemistry for monocytes/macrophages (Mo/Mphi, ED-1), myofibroblasts [alpha-smooth muscle actin (alpha-SMA)], collagen III and IV, matrix metalloproteinase-2 (MMP-2) and proliferating cell nuclear antigen (PCNA) and terminal dUTP nick end labeling (TUNEL) were performed. RESULTS: Kidney weights of postischemic animals were increased at all time points (postischemic to controls, 1.47 +/- 0.21 to 0.94 +/- 0.12 g at day 8; 1.49 +/- 0.20 to 1.27 +/- 0.13 g at day 64; and 1.86 +/- 0.1 to 1. 24 +/- 0.2 g at day 180). Serum creatinine values increased to 0.42 +/- 0.10 mmol/L at day 2 but returned to control levels by day 8 (0. 05 mmol/L). Glomerular collagen IV was decreased from 2 to 16 days postischemia, which was accompanied by an increase in MMP-2. The fractional area of the interstitium was greatest at day 8 (19.55 +/- 0.91% compared with day 0 at 8.08 +/- 0.27%), with a second increase observed at day 180 (16.61 +/- 0.70%). Interstitial Mo/Mphi increased postischemia from days 2 through 8 (8.84 +/- 2.12 to 133. 32 +/- 14.04 per 0.91 mm2) and then decreased. Myofibroblasts proliferated locally (PCNA double labeling was demonstrated), and increased numbers were found from days 2 through 16 (maximal at day 8, 26.96 +/- 3.04%, compared with day 0, 0.88 +/- 0.11%). In the postischemic groups, collagen IV increased to day 8 (20.84 +/- 1. 30%), but then decreased to below control values at day 64 (2.22 +/- 0.15%) before returning to normal by day 180. Interstitial collagen III increased to 8 days (0.45 +/- 0.07% to 2.55 +/- 0.36%) and then decreased to control levels by day 32, but showed a marked increase to approximately 6% at days 64 and 180. Cellular proliferation (PCNA) was maximal at days 2 and 4 (affecting tubule cells and myofibroblasts but not macrophages). Apoptosis was maximal at day 8 (in both interstitial and tubule cells) in the postischemic groups. CONCLUSION: Marked changes in the accumulation of Mo/Mphi, MF, and collagen IV were found in this model of ischemic acute renal failure. The reversibility of functional and structural changes is in marked contrast to that found in progressive disease. The increases observed for collagen III at 64 and 180 days postischemia suggest that in the long term, however, further chronic structural changes may be observed. PMID- 10844608 TI - Tight junction proteins ZO-1, ZO-2, and occludin along isolated renal tubules. AB - BACKGROUND: Tight junctions play a critical role in tubular function. In mammalian kidney, the transepithelial electrical resistance and the complexity of the tight junction increase from the proximal to the collecting tubule. The differential expression of three tight junction proteins, ZO-1, ZO-2, and occludin, along isolated rabbit renal tubules is examined in this article. METHODS: Microdissected rabbit renal tubules were processed for immunofluorescence detection of ZO-1, ZO-2, and occludin. The quantitation of these proteins was done by Western blot determinations in Percoll isolated tubules. RESULTS: ZO-1 stained cell boundaries independently of the identity of the tubule. However, the amount found in distal segments was significantly higher than that expressed in proximal regions. ZO-2 in the proximal region was found diffusely distributed in the cytoplasm, with faint staining at cell borders, while a clear signal at cell perimeters was detectable from the Henle's loop to collecting tubules. Nuclear staining of ZO-2 was found along the whole nephron. The presence of occludin at the proximal region was faint and discontinuous, while its expression in the more distant portions was conspicuous. The quantity of ZO-2 and occludin present at the distal region was significantly higher compared with the proximal segment. CONCLUSIONS: The distribution of ZO-1, ZO-2, and occludin follows the increase in junction complexity encountered in renal tubules. The amount of the three proteins found in proximal and distal segments is significantly higher in the latter. PMID- 10844609 TI - Oxalate-induced ceramide accumulation in Madin-Darby canine kidney and LLC-PK1 cells. AB - BACKGROUND: Oxalate exposure produces oxidant stress in renal epithelial cells leading to death of some cells and adaptation of others. The pathways involved in these diverse actions remain unclear, but appear to involve activation of phospholipase A2 (PLA2) and redistribution of membrane phospholipids. The present studies examined the possibility that oxalate actions may also involve increased accumulation of ceramide, a lipid-signaling molecule implicated in a variety of pathways, including those leading to apoptotic cell death. METHODS: Ceramide accumulation was examined in renal epithelial cells from pig kidney (LLC-PK1 cells) and from dog kidney [Madin-Darby canine kidney (MDCK cells)] using the diacylglycerol kinase assay. Sphingomyelin degradation was assessed by monitoring the disappearance of 3H-sphingomyelin from cells that had been prelabeled with [3H]-choline. The effects of oxalate were compared with those of other oxidants (peroxide, xanthine/xanthine oxidase), other organic acids (formate and citrate), and a known activator of sphingomyelinase in these cells [tumor necrosis factor alpha (TNF-alpha)]. Separate studies determined whether oxalate-induced accumulation of ceramide could be blocked by pretreatment with antioxidants [Mn (III) tetrakis (1-methyl-4-pyridyl) porphyrin (Mn TMPyP, a superoxide dismutase mimetic) or N-acetylcysteine (NAC; an antioxidant)], with an inhibitor of ceramide synthase [fumonisin B1 (FB1)] or with an inhibitor of PLA2 [arachidonyl trifluoromethylketone (AACOCF3)]. RESULTS: Oxalate exposure produced a significant time- and concentration-dependent increase in cellular ceramide. A reciprocal decrease in 3H-sphingomyelin was observed under these conditions. Increases in cellular ceramide levels were also observed after treatment with other oxidants (hydrogen peroxide, and xanthine/xanthine oxidase), activators of sphingomyelinase (TNF-alpha), exogenous sphingomyelinase, or arachidonic acid. Formate produced similar (albeit smaller) effects, and citrate did not. The oxidant-induced increases in ceramide were attenuated by pretreatment with NAC (a glutathione precursor) and MnTMPyP (a superoxide dismutase mimetic), suggesting a role for cellular redox states. The oxalate-induced increase in ceramide was also attenuated by pretreatment with AACOCF3, suggesting a role for PLA2. Pretreatment with FB1 produced a small but statistically insignificant attenuation of the response to oxalate. CONCLUSIONS: Oxalate exposure produces a marked accumulation of ceramide in renal epithelial cells by a process that is redox sensitive and mediated in part by activation of PLA2. Since cellular sphingomyelin decreased as ceramide increased, it seems likely that oxalate actions are mediated, at least in part, by an increase in sphingomyelinase activity, although alterations in ceramide synthase are also possible. Further study is required to define the steps involved in oxalate actions and to determine the extent to which ceramide signaling mediates oxalate actions. PMID- 10844610 TI - Proteinase-3 mRNA expressed by glomerular epithelial cells correlates with crescent formation in Wegener's granulomatosis. AB - BACKGROUND: Wegener's granulomatosis (WG) is characterized by systemic vasculitis with crescentic glomerulonephritis (CGN) and circulating autoantibodies directed against neutrophil cytoplasmic antigens (ANCA). Proteinase 3 (PR-3), a neutral serine proteinase in neutrophils implicated in the growth control of myeloid cells, has been identified as the target antigen for ANCA in WG. Since the kidneys are frequently involved in WG, we studied the in situ expression of PR-3 by renal parenchymal cells. METHODS: We assessed the expression of PR-3 in kidney biopsies of 15 patients with WG by immunohistochemistry (IHC) and in situ hybridization (ISH). Normal kidney tissue served as the control. RESULTS: We detected PR-3 mRNA and PR-3 protein in distal tubular epithelial cells (TECs) and glomerular epithelial cells (GECs) in normal kidney tissue and in CGN. Furthermore, a strong glomerular PR-3mRNA expression restricted to the site of cellular crescents was detected in patients with WG. The analysis of 144 glomeruli with cellular or sclerotic crescents revealed a positive correlation of glomerular PR-3mRNA expression with the percentage of cellular crescents per glomerulus. The capability of human TECs and GECs to synthesize PR-3 was confirmed by Northern blot and ISH on cultured cells. CONCLUSION: These data provide evidence that nonhematopoetic renal parenchymal cells express PR-3 and that glomerular expression of PR-3 is associated with crescent formation in WG. Our findings suggest that renal parenchymal cells may directly be involved in the pathogenesis of CGN in WG. PMID- 10844611 TI - Renal response to repetitive exposure to heme proteins: chronic injury induced by an acute insult. AB - BACKGROUND: Renal diseases are conventionally classified into acute and chronic disorders. We questioned whether acute, reversible, renal insults may be induced to incite a chronic scarring process, employing as an acute insult the glycerol model of heme protein-induced renal injury. METHODS: Rats were subjected to weekly injections of hypertonic glycerol for up to six months. Renal function was serially determined, and the effect of such insults on renal histology and renal expression of collagen and fibrogenic cytokines was assessed. RESULTS: After the first injection of glycerol, which, expectedly, induced a prompt fall in the glomerular filtration rate (GFR), subsequent injections encountered a remarkable renal resistance in that the fall in GFR was markedly blunted. This resistance to acute decline in renal function in rats subjected to repetitive injections of glycerol was accompanied by less necrosis and apoptosis of renal tubular epithelial cells after such injections. The attenuation in the fall in GFR in response to repetitive exposure to glycerol-induced heme protein injury was maintained for up to six months. A progressive decline in GFR appeared after three months and was accompanied by histologic tubulointerstitial injury, the latter assessed at six months. These kidneys demonstrated up-regulation of collagen I, III, and IV in conjunction with increased expression of the oxidant inducible, chemotactic cytokine, monocyte chemoattractant protein-1 (MCP-1), and the oxidant-inducible, fibrogenic cytokine, transforming growth factor-beta1 (TGF beta1). The exposure of the kidney to a single injection of hypertonic glycerol increased the expression of both cytokines some three to five days following this exposure, while the exposure of NRK 49F cells in culture to an iron-dependent model of oxidative stress also increased expression of TGF-beta1 and collagen mRNAs. CONCLUSIONS: We conclude that this nephrotoxic insult, repetitively administered, encounters a resistance in the kidney such that the expected fall in GFR does not occur. However, with time, such resistance is accompanied by a decrease in GFR, the latter associated with chronic tubulointerstitial disease. Thus, a long-term cost is exacted, either along with, or as a consequence of, such resistance. We suggest that chronic up-regulation of such oxidant-inducible genes such as TGF-beta1 and MCP-1 contributes to tubulointerstitial disease, and iron-mediated oxidative stress may directly induce TGF-beta1. PMID- 10844612 TI - Transforming growth factor-beta isoforms and glomerular injury in nephrotoxic nephritis. AB - BACKGROUND: Transforming growth factor-beta has three main isoforms (TGF-beta1, TGF-beta2, and TGF-beta3) that have distinct but overlapping functions in immunity, inflammation, and tissue repair. TGF-beta1 has been implicated in progressive renal scarring, but the roles of TGF-beta2 and TGF-beta3 are less clear. The purpose of this study was to characterize the expression of all three isoforms in nephrotoxic nephritis (NTN) in rats and to determine the effect of TGF-beta3 infusions on injury because of its reported combined anti-inflammatory and antifibrotic effects. METHODS: TGF-beta1, TGF-beta2, and TGF-beta3 expression was analyzed by immunohistochemistry and RNase protection assays. TGF-beta3 was administered by osmotic minipumps at 2 microg/day, a dose shown to alter glomerular macrophage function in vivo. Injury was assessed morphologically and functionally. RESULTS: The three TGF-beta isoforms showed a different distribution in normal rats and after the induction of nephritis. TGF-beta1 was only detected in glomeruli of the most severely nephritic rats. TGF-beta2 was found in glomerular neutrophils, whereas damaged podocytes expressed TGF-beta3. Infusions of TGF-beta3 did not reduce proteinuria over seven days after the induction of nephritis. They did, however, have a profound effect on glomerular macrophage number (7.76 +/- 4.1 in treated rats vs. 14.4 +/- 4.7 in controls, P < 0.02). The numbers of class II-positive macrophages were similar in the two groups, whereas class II-negative macrophages infiltrating glomeruli were significantly decreased (4.06 +/- 3.1 vs. 9.1 +/- 4.4, P < 0.02). TGF-beta did not influence the amount of glomerular matrix. CONCLUSIONS: TGF-beta isoforms have different expressions and presumptively different roles in NTN. The infusion of pharmacological doses of TGF-beta3 has profound effects on macrophages infiltrating nephritic glomeruli and reveals marked heterogeneity of infiltrating macrophages. PMID- 10844613 TI - Redistribution of cytoplasmic VEGF to the basolateral aspect of renal tubular cells in ischemia-reperfusion injury. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) mRNA and protein expression are increased by hypoxia in a variety of cell types and organs. In the kidney, however, chronic hypoxia does not up-regulate VEGF mRNA. This suggests that VEGF may be regulated by unique mechanisms in the kidney. METHODS: Unilateral ischemia was induced in rats by vascular cross-clamping (40 min) followed by reperfusion (0, 20, 40, and 80 min). The distribution of VEGF protein was determined by immunohistochemical staining and Western blotting. mRNA was detected by Northern blotting and semiquantitative reverse transcription-polymerase chain reaction (RT PCR). Immunohistochemical staining for VEGF was verified using two VEGF antibodies. To further substantiate the immunohistochemical findings, laser scanning confocal fluorescence microscopy was used to demonstrate the distribution of VEGF protein in rat renal tubular epithelial cells (NRK52-E) subjected to hypoxia (40 min) and re-oxygenation (0, 5, 20, 40 and 80 min). RESULTS: Normal kidneys showed diffuse immunohistochemical staining for VEGF in all tubules of the renal cortex and medulla. Following ischemia, staining demonstrated a prominent shift of cytoplasmic VEGF to the basolateral aspect of tubular cells with both VEGF antibodies. The distribution of cytoplasmic VEGF returned to normal following 40 and 80 minutes of reperfusion. Western blots of cytoplasmic samples from ischemic kidneys reperfused for 0 and 20 minutes showed decreased levels of VEGF164 compared with normal (P < 0.01). VEGF164 and VEGF188 levels in the membrane fraction showed no change. Northern blots and semiquantitative RT-PCR showed no significant up-regulation of VEGF mRNA or change in the splice pattern. NRK52-E cells subjected to hypoxia and re oxygenation for 0 and 5 minutes showed increased staining for VEGF compared with normal, with prominent VEGF staining at the periphery of the cell, similar to the appearance in ischemic kidneys. VEGF staining became more diffuse with further re oxygenation. CONCLUSION: Although synthesis of VEGF mRNA and protein is not increased during ischemia reperfusion injury, pre-existing VEGF in the tubular cell cytoplasm redistributes to the basolateral aspect of the cells. These data suggest that the kidney may have evolved unique patterns of VEGF regulation to cope with acute hypoxia. PMID- 10844614 TI - Mechanism of regulation of Na+ transport by angiotensin II in primary renal cells. AB - BACKGROUND: Angiotensin II (Ang II) has a dose-dependent, biphasic effect on the activity of the Na+/H+ antiport system in the renal proximal tubule (RPT). The aim of the present study was to further delineate the signaling pathways involved in Ang II action. METHODS: To examine Ang II signaling, 22Na+ uptake studies were conducted with a primary rabbit RPT cell culture system. The activation of phospholipase A2 (PLA2) was assessed by measuring the release of [3H]-arachidonic acid (AA), and changes in intracellular calcium levels were determined by means of confocal microscopy. RESULTS: Low dosages of Ang II (<10-10 mol/L) stimulated Na+ uptake, whereas high dosages of Ang II (>10-10 mol/L) inhibited Na+ uptake. Ang II (>10-10 mol/L) also caused an increase in AA release associated with an increase in intracellular calcium. Not only did exogenous AA inhibit Na+ uptake, but two PLA2 inhibitors (mepacrine and AACOCF3) blocked the Ang II-mediated inhibition of Na+ uptake. However, the cytochrome P450-dependent epoxygenase inhibitor econazole also blocked the Ang II-induced inhibition of Na+ uptake. Inhibition of Na+ uptake was obtained by the metabolic product of the epoxygenase 5,6-EET. In turn, the inhibitory effect of 5,6-EET was blocked by indomethacin. CONCLUSIONS: The results indicate the involvement of a calcium-dependent PLA2 in mediating the inhibitory effect of Ang II on Na+ uptake. The AA, which is released following PLA2 activation, acts indirectly, through its own metabolism, via a cytochrome P450 epoxygenase pathway and ultimately cyclooxygenase itself. PMID- 10844615 TI - Culture of vascular smooth muscle cells from small arteries of the rat kidney. AB - BACKGROUND: In contrast to arterioles, small arteries appear to be the preferential site of renal vascular smooth muscle cell (VSMC) proliferation under pathophysiological conditions. To date, techniques have been described to isolate renal arterioles and to culture VSMCs. The aim of the present study was to develop a method of culturing VSMCs from isolated small arteries of the rat kidney and to characterize their growth as compared with that of aortic VSMCs. METHODS: Renal vascular trees were isolated from kidneys of male Wistar rats by a sieving technique. VSMCs were grown from explants of collagenase-treated renal vascular trees and thoracic aorta. Growth curves and proliferation of renal and aortic VSMCs in response to fetal bovine serum (FBS) were compared by determination of cell number and DNA synthesis, measured as incorporation of 5 bromo-2'-deoxyuridine. RESULTS: Renal vascular trees consisted mainly of small arteries with a diameter of 80 to 400 microm (interlobar and arcuate arteries). As compared with total kidney or renal cortex, alkaline phosphatase activity was decreased by 81%, and vasopressin (10 micromol/L) was unable to stimulate adenylyl cyclase in renal vascular trees, indicating little tubular contamination. A homogenous population of spindle-shaped cells was cultured from renal vascular trees, which grew in a hill-and-valley pattern and stained positively for smooth muscle alpha-actin, according to the characteristics of VSMC phenotype. Renal VSMCs proliferated more slowly than aortic VSMCs and reached the plateau of growth at about half of the cell density of aortic VSMCs. Furthermore, proliferation of renal VSMCs depended more heavily on FBS concentration, since about threefold higher concentrations of FBS were needed for renal VSMCs to multiply at the same rate and to similarly stimulate DNA synthesis as compared with aortic VSMCs. CONCLUSIONS: We present a method to culture renal VSMCs from small arteries of the rat kidney, which possess distinct growth characteristics as compared with aortic VSMCs. PMID- 10844616 TI - In vivo effects of diadenosine polyphosphates on rat renal microcirculation. AB - BACKGROUND: Diadenosine polyphosphates (APXA) are vasoactive nucleotides that elicit effects via purinoceptors. Recent data suggest differential effects of APXA on kidney vasculature. METHODS: The in vivo effects of AP3A, AP5A, and adenosine on renal microvessels and the role of purinoceptors were investigated by the application of agonists to the hydronephrotic rat kidney and preincubation with respective antagonists. RESULTS: The addition of the agonists (10-7 mol/L up to 10-4 mol/L) resulted in a concentration-dependent transient vasoconstriction [interlobular artery (ILOB): adenosine 30 +/- 7%, N = 7, AP3A 35 +/- 10%, N = 5; AP5A 66 +/- 19%, N = 5; 10-5 mol/L each] lasting up to one minute, followed by a concentration-dependent vasodilation (ILOB: adenosine 10 +/- 3%, N = 6; AP3A 19 +/- 4%, N = 5; AP5A 12 +/- 5%, N = 6; 10-5 mol/L each). In ILOB and in the afferent arteriole (AFF), the constrictory effects of AP5A were more pronounced than those of AP3A and adenosine. In the efferent arteriole (EFF), vascular tone was only slightly affected by all agonists. The dilatory potency was comparable for all agonists in ILOB and EFF. No significant vasodilation occurred in AFF. The application of the selective A1 receptor antagonist DPCPX (10-5 mol/L) completely abolished the adenosine-induced vasoconstriction, whereas the A2 receptor antagonist DMPX and the P2 purinoceptor antagonists PPADS and A3P5P (all 10-5 mol/L) did not affect adenosine-induced constriction. The AP3A-induced constriction was abolished by DPCPX and was partially inhibited by PPADS. The constriction induced by AP5A was less sensitive to DPCPX but more sensitive to PPADS. In ILOB and EFF, DMPX or A3P5P abolished dilation after the addition of the agonists. The dilation after AP5A was not significantly reduced. In AFF, no significant dilation was observed with these agonists alone, but it was clearly visible in the presence of DPCPX or PPADS. CONCLUSIONS: APXA evoke transient constrictions in vessels of the hydronephrotic rat kidney, which are mediated by A1 and P2 purinoceptors. The length of the phosphate chain determines the degree of vasoconstriction and the extent to which the substances exert effects on the P2 purinoceptor subtypes. ILOB and AFF are more potently affected by APXA than EFF. Afferent vasodilation is partially overridden by sustained vasoconstriction. PMID- 10844617 TI - Nocturnal blood pressure and 24-hour pulse pressure are potent indicators of mortality in hemodialysis patients. AB - BACKGROUND: Cardiovascular (CV) complications are the leading cause of mortality in hemodialysis patients. The role of arterial hypertension on the prognosis of CV in hemodialysis patients is not as clear as in the general population. The purpose of this study was to investigate the prognostic role of ambulatory blood pressure (BP) on CV mortality in treated hypertensive hemodialysis patients. METHODS: Fifty-seven treated hypertensive hemodialysis patients (56.87 +/- 16.22 years, 30 men) were prospectively studied. All patients initially underwent an ambulatory BP monitoring between two dialysis sessions. The outcome event studied was CV death; kidney transplantation and deaths not related to CV disease were censored. RESULTS: The duration of follow-up was 34.4 +/- 20.39 months, during which 10 CV and 8 non-CV fatal events occurred. In the 10 patients who died from CV complications, age, previous CV events, ambulatory systolic BP, ambulatory pulse pressure (PP), and life-long smoking level were significantly higher, and the office diastolic BP was lower at the time of inclusion than in those who did not die from CV complications (N = 47). Based on Cox analysis and after adjustment for age, sex, and previous CV events, a low office diastolic BP [relative risk (RR) 0.49, 95% CI, 0.25 to 0.93, P = 0.03], an elevated 24-hour PP (RR 1.85, 95% CI, 1.28 to 2.65, P = 0.009), and an elevated nocturnal systolic BP (RR 1.41, 95% CI, 1.08 to 1.84, P = 0.01) were predictors of CV mortality (RR associated with a 10 mm Hg increase in BP and in PP). CONCLUSION: This study demonstrates that nocturnal BP and 24-hour PP are independent predictors of CV mortality in treated hypertensive hemodialysis patients. Randomized trials are needed to investigate whether nocturnal BP and 24-hour PP are superior to office BP as targets for antihypertensive therapy in this high-risk group. PMID- 10844618 TI - Effect of insulin and heparin on glucose-induced vascular damage in cell culture. AB - BACKGROUND: Clinical trials have shown that tight glycemic control reduces the risk of diabetic microvascular complications, namely retinopathy, nephropathy, and neuropathy. The mechanism of these microvascular complications is not yet fully elucidated. The present study describes the effect of different concentrations of glucose on vascular endothelial cells (ECs) and vascular smooth muscle cells (VSMCs) in cell culture. Our objective was to shed some light through this biological study on the mechanism and prevention of diabetic microvascular complications. METHODS: ECs and VSMCs were treated with 5 mmol/L (90 mg/dL) or 30 mmol/L (540 mg/dL) D-glucose or D-glucose plus insulin or D glucose plus insulin and heparin in culture. ECs were studied with light microscopy (LM), scanning electron microscopy (SEM), and transmission electron microscopy (TEM) for surface changes. The cultured ECs were treated with vimentin antibodies and VSMCs with actin antibodies for immunoflourescence microscopy (IFM) study. Endothelin-1 (ET-1) assay was done on ECs culture medium using enzyme-linked immunosorbent assay (ELISA). RESULTS: LM, SEM, and TEM of ECs treated with a physiological concentration (90 mg/dL) of D-glucose appeared the same as control. However, LM and SEM of ECs treated with a high concentration of D-glucose (540 mg/dL) showed pronounced intercellular gaps. This finding was further confirmed by TEM study. These gaps were minimally or not at all discernible when insulin, heparin, or a combination of both was added to the culture medium. IFM showed increased vimentin expression with a high concentration of D-glucose. Vimentin expression was attenuated with the addition of insulin or heparin in the medium and more markedly with combined insulin and heparin. Significant correlations were obtained between glucose levels, vimentin expression, and ET-1 levels. The higher the glucose level, the higher is the ET-1 production and the greater vimentin expression in ECs. Cultured VSMCs treated with a high concentration of D-glucose showed enhanced actin expression. Actin expression was blunted with the addition of insulin or heparin in the culture medium. CONCLUSIONS: These biological findings indicate the salutary effect of insulin or insulin and heparin in the mitigation of vascular disorganization caused by a high concentration of D-glucose. PMID- 10844619 TI - Lipopolysaccharide impairs endothelial nitric oxide synthesis in rat renal arteries. AB - BACKGROUND: Impaired endothelium-dependent vasodilation may contribute to hypoperfusion and failure of abdominal organs, including the kidneys during endotoxin or septic shock. In this study, the short-term (2 h) effects of bacterial lipopolysaccharide (LPS) on endothelium-dependent vasodilation in rat renal and superior mesenteric arteries were documented. METHODS: Rat renal and mesenteric arteries were dissected and exposed in vitro to LPS for two hours. The effects of LPS on vascular reactivity were determined and compared with time matched controls. Endothelial nitric oxide (NO) release was determined using an NO microsensor in adjacent vessel segments. RESULTS: LPS impaired maximal acetylcholine (ACh)-induced endothelium-dependent vasodilation in renal arteries (62.5 +/- 8.8% vs. 34.4 +/- 7.5% in controls and LPS-exposed arteries), but not in mesenteric arteries. LPS did not alter the sensitivity of renal arteries to exogenous NO. ACh-dependent vasodilation was abolished after blocking NO synthesis with 10-4 mol/L L-NA in control and LPS-incubated renal arteries. When compared with controls, NO release induced by ACh and the receptor-independent calcium ionophore A23187 was significantly decreased (P < 0.05) in LPS-exposed renal segments and was fully abolished in endothelium-denuded segments, indicating that LPS attenuated receptor-dependent as well as receptor-independent endothelial NO release. In contrast, ACh- and A23187-induced NO release was normal in LPS-exposed mesenteric arteries. CONCLUSIONS: These results indicate that LPS-induced selective impairment of ACh-induced endothelium-dependent relaxation in rat renal arteries is caused by decreased endothelial NO release. This may contribute to the propensity for acute renal failure during septic shock. PMID- 10844620 TI - Renal blood flow measurement by positron emission tomography using 15O-labeled water. AB - BACKGROUND: Only few noninvasive methods have the potential to quantitate renal blood flow (RBF) in humans. Positron emission tomography (PET) is a clinical imaging method that can be used to measure the tissue blood flow noninvasively. The purpose of this study was to validate PET measurement of RBF using 15O labeled water (H215O), a tracer that allows repeated measurements at short time intervals. METHODS: RBF was measured in six pigs by PET and by radioactive microspheres (MS). Three measurements were performed in each pig at baseline (BL), during vascular expansion and dopamine infusion (DA; 20 microg. kg-1. min-1 intravenously), and during angiotensin II (Ang II) infusion (50 ng. kg-1. min-1 intravenously). RBF was estimated from aortic and renal tracer kinetics using a model adapted from the blood flow model described by Kety and Smith. RESULTS: PET and MS values correlated strongly (y = 0.79x + 42, r = 0.93, P < 0.0001) over the RBF range from 100 to 500 mL. min-1. 100 g-1. Pharmacologically induced changes were significant and were measured equally well by PET and MS: 38 and 39%, respectively, below BL (P < 0.005 and P < 0.05) under Ang II, and 47 and 48%, respectively, above BL (P < 0.005 and P < 0.01) under DA. A Bland and Altman representation showed a low average difference of -17 +/- 45 mL. min-1. 100 g-1 (mean +/- SD). CONCLUSION: To our knowledge, this study provides the first validation of RBF measurement by PET using H215O over a large range of RBF values (100 to 500 mL. min-1. 100 g-1), which correspond to RBF values in both healthy subjects and in patients suffering from chronic renal failure. PMID- 10844621 TI - Age, blood pressure and smoking effects on chronic renal failure in primary glomerular nephropathies. AB - BACKGROUND: Smoking is a known risk factor for renal damage in diabetic patients, but its relationship to other renal diseases is less clear. To assess its effect in primary glomerular nephropathy (GN), we used data from a case control study designed to assess several environmental risk factors. METHODS: This study included 295 biopsy-proven GN cases, 80 membranous nephropathy, 116 IgA nephropathy, and 99 nephrotic syndrome with either minimal change nephropathy or focal segmental hyalinosis, and 242 matched hospital controls, with diseases unrelated to smoking. Subjects were interviewed about their smoking history. Chronic renal failure (CRF), defined by serum creatinine> 150 micromol/L, was present in 74 cases (57 men and 17 women). Logistic regression was used to estimate odds ratios (ORs) adjusted for age and social class. RESULTS: In men, the percentage of ever-smokers did not differ between GN cases (60%) and controls [65%, OR = 0.9 (95% Confidence Interval 0.6-1.4)], but was significantly higher among cases with CRF (75%) than those without [55%, OR = 2.4 (1.2-4.5)]. Dose effect relationships were observed with both the daily and cumulative dose; this relationship was stronger in the comparison of cases with CRF and those without CRF than in the comparison of cases with CRF with controls: OR = 1.9 versus 1.3 (20 cigarettes/day); OR = 1.9 versus 1.4 (15 pack years). Interactions between age, hypertension, and smoking were observed in the risk of CRF: smoking was significantly related to CRF among cases who were older than 40 years and/or hypertensive, but not among those cases younger than 40 or normotensive. The results did not significantly differ among the three histologic types. No relationship was shown between smoking and CRF in women. CONCLUSION: This study provides additional support for the hypothesis that smoking is related to GN severity, particularly in the at-risk groups of men older than 40 and/or hypertensive patients. These findings should be corroborated by further observations in other populations. PMID- 10844622 TI - Randomized double-blind trial of oral essential amino acids for dialysis associated hypoalbuminemia. AB - BACKGROUND: Hypoalbuminemia is associated with substantial morbidity and mortality in dialysis patients. METHODS: Subjects with a mean three-month prestudy serum albumin of 3.8 g/dL or less and who demonstrated >/=90% compliance during a two-week run-in period were randomized to 3.6 g of essential amino acids (EAAs) or placebo three times daily with meals for three months. Randomization was stratified by dialysis modality and by severity of the hypoalbuminemia. The primary study outcome was change in the average of three monthly serum albumin measurements between baseline and follow-up. RESULTS: Fifty-two patients were randomized; 47 patients (29 hemodialysis and 18 peritoneal dialysis) met the predetermined primary analysis criteria. The mean compliance rates averaged 75, 70, and 50% at months 1, 2, and 3, respectively, and were similar for EAAs and placebo. Serum albumin in the hemodialysis patients, EAA versus placebo, improved [(mean +/- SE) 0.22 +/- 0.09 g/dL, P = 0.02]. Changes in peritoneal dialysis patients were not significant (0.01 +/- 0.15 g/dL), but approached significance for the total study group (0.14 +/- 0.08 g/dL, P = 0.08). Patients in the very low albumin strata (<3.5 g/dL) improved more than those in the low albumin strata (3.5 to 3.8 g/dL, P < 0.01). There was a significant correlation (r = 0.83, P = 0.001) within the hemodialysis EAA group between the baseline C-reactive protein level and improvement in serum albumin. Improvements were also seen in grip strength and SF-12 mental health score, but not in serum amino acid levels, SF-12 physical health score, or anthropometric measurements. CONCLUSIONS: Oral EAAs induce a significant improvement in the serum albumin concentration in hemodialysis but not peritoneal dialysis subjects. Further study of their long term effects on morbidity and mortality is warranted. PMID- 10844623 TI - Can we improve early mortality in patients receiving renal replacement therapy? AB - BACKGROUND: Approximately one in eight patients with end-stage renal disease (ESRD) die within the first three months of starting renal replacement therapy (RRT). We investigated which factors might improve this early mortality. METHODS: We performed a prospective nationwide study of all patients commencing RRT for ESRD in Scotland over one year. Patients were classified according to how they presented to start RRT, their burden of comorbid diseases, access prepared for dialysis, and duration of care by a nephrologist prior to commencing RRT. Those factors most strongly associated with death within 90 days of commencing treatment were determined by logistic regression analysis. RESULTS: Patients with an acute unexpected element to their presentation for RRT had early mortality rates between 6.0 and 8.9 times greater than those who commenced RRT electively after a period of care from a nephrologist. Patients in high and medium comorbidity risk groups had early mortality rates of 4.7 and 2.2 times greater than those in the low-risk group. Low serum albumin had a significant association with early death. Patients who progressed steadily to ESRD, who had a planned start to dialysis, and who had mature access were 3.6 times more likely to survive beyond three months than those with no access; they were, however, also younger with less comorbidity. CONCLUSIONS: The factors principally associated with early mortality are nonelective presentation for RRT, comorbid illness, and low serum albumin. Patients cared for by a nephrologist before requiring RRT who have mature access have better short-term survival than those without access. They are also younger with less comorbidity. It may be possible to improve short term survival in this "unplanned" group if referred early to facilitate reducing cardiovascular risk factors and preparation for RRT. PMID- 10844624 TI - Computer simulations of ultrafiltration profiles for an icodextrin-based peritoneal fluid in CAPD. AB - BACKGROUND: The three-pore model of peritoneal transport has the ability to predict ultrafiltration (UF) profiles rather accurately, even when high molecular weight (MW) solutes are employed as osmotic agents in continuous ambulatory peritoneal dialysis (CAPD). In the present simulations, we wanted to assess, for various theoretical perturbations, the UF properties of a peritoneal dialysis (PD) solution with an osmotic agent having an average MW of 20 kD and a "number average MW" of 6.2 kD, which is similar to that of icodextrin (ICO). METHODS: For a PD solution containing a completely monodispersed 20 kD MW osmotic agent, the degree of UF modeled is much higher than that reported for ICO. Hence, to model the behavior of ICO, we subdivided the ICO molecules into eight or more different MW size fractions. For simulations using six or eight subfractions, we obtained an excellent fit of simulated to reported UF data. More dispersed solutions produced UF profiles similar to that with eight fractions. RESULTS: A 2.05 L 7.5% ICO PD solution, despite being slightly hypotonic, yielded a UF volume of nearly 600 mL in 12 hours, modeled for patients not previously exposed for ICO. After nine hours, the UF volume exceeded that produced by 3.86% glucose. The UF rate and volumes increased in proportion to (1) the ICO concentration, (2) the peritoneal surface area, and (3) the peritoneal UF coefficient, but was almost insensitive to increases in the instilled fluid volume. Simulated for patients previously exposed to ICO, having steady-state plasma concentrations of ICO degradation products, the predicted UF volume at 12 hours was reduced to approximately 400 mL. CONCLUSION: Employing the three-pore model of peritoneal transport and taking into account the polydispersed nature of ICO, it was possible to accurately computer simulate the UF profiles of ICO in accordance with reported data. The simulations suggest an advantage of using ICO in patients with type I UF failure, where UF with a high-MW osmotic agent will exceed that seen in patients not showing UF failure who are on glucose-based PD solutions. PMID- 10844625 TI - Comparative hospitalization of hemodialysis and peritoneal dialysis patients in Canada. AB - BACKGROUND: Most comparisons of hemodialysis (HD) and peritoneal dialysis (PD) have used mortality as an outcome. Relatively few studies have directly compared the hospitalization rates, an outcome of perhaps equal importance, of patients using these different dialysis modalities. METHODS: Eight hundred twenty-two consecutive patients at 11 Canadian institutions with irreversible renal failure had an extensive assessment of comorbid illness and initial mode of dialysis collected prospectively immediately prior to starting dialysis therapy. The cohort was assembled between March 1993 and November 1994. The mean follow-up was 24 months. Admission data were used to compare hospitalization rates in HD and PD. RESULTS: Thirty-four percent of patients at baseline and 50% at three months used PD. Twenty-five percent of HD and 32% of PD patients switched dialysis modality at least once after their first treatment (P = NS). Nine percent of HD patients and 30% of PD patients switched modality after three months (P < 0. 001). Total comorbidity was higher in HD patients at baseline (P < 0. 001) and at three months (P = 0.001). The overall hospitalization rate was 40.2 days per 1000 patient days after baseline and 38.0 days per 1000 patient days after three months. When an adjustment was made for baseline comorbid conditions, patients on PD had a lower rate of hospitalization in intention-to-treat analysis according to the type of dialysis in use at baseline (RR 0.85, 95% CI, 0.82 to 0.87, P < 0.001), but a higher rate according to the type of dialysis in use three months after study entry (RR 1.31, 95% CI, 1.27 to 1.34, P < 0.001). In analyses based on the amount of time actually spent on each treatment modality, PD was associated with a higher rate of hospitalization when analyzed according to the type of dialysis in use at baseline (RR 1.10, 95% CI, 1.07 to 1.13, P < 0.001) and according to the type of dialysis in use three months after study entry (RR 1.26, 95% CI, 1.23 to 1.30, P < 0.001). CONCLUSIONS: Conclusions regarding comparative hospitalization rates are heavily dependent on the analytic starting point and on whether intention-to-treat or treatment-received analyses are used. When early treatment switches are accounted for, HD is associated with a lower rate of hospitalization than PD, but the effect is modest. PMID- 10844626 TI - Physical activity levels in patients on hemodialysis and healthy sedentary controls. AB - BACKGROUND: Patients on dialysis have reduced exercise tolerance compared with age-matched sedentary controls. The reasons for this debility have not been fully elucidated, but physical inactivity could be a contributing factor. The purpose of the current study was to determine whether patients on hemodialysis are less active than healthy sedentary controls and to explore clinical correlates of physical activity level in a group of hemodialysis patients. METHODS: Thirty-four hemodialysis patients and 80 healthy sedentary individuals participated in the study. Physical activity was measured for seven days with a three-dimensional accelerometer and with an activity questionnaire. RESULTS: Vector magnitude values from the accelerometer for the dialysis and control subjects were 104,718 +/- 9631 and 161,255 +/- 6792 arbitrary units per day, respectively (P < 0.0001, mean +/- SEM). The estimated energy expenditure values derived from the questionnaire were 33.6 +/- 0.5 kcal/kg/day and 36.2 +/- 0.5 kcal/kg/day (P = 0.002). The difference between patients on dialysis and controls increased with advancing age. Among the dialysis subjects, some measures of nutritional status correlated with physical activity level, including serum albumin concentration (r = 0.58, P = 0.003), serum creatinine concentration (r = 0.37, P = 0. 03), and phase angle derived from bioelectrical impedance analysis (r = 0.40, P = 0.02). CONCLUSIONS: Patients on hemodialysis are less active than healthy sedentary controls, and this difference is more pronounced among older individuals. There is an association between the level of physical activity and nutritional status among patients on dialysis. These findings are of great concern, given the trend toward increasing age in incident dialysis patients and the well-known association between inactivity and increased mortality in the general population. PMID- 10844627 TI - Mechanisms for the formation of glycoxidation products in end-stage renal disease. AB - BACKGROUND: Advanced glycation end products (AGEs) accumulate on tissue and plasma proteins in patients with renal failure far in excess of normal aging or diabetes. The aim of these studies was to elucidate the nature of the precursors and the pathways that lead to an accelerated formation of two structurally identified AGEs [pentosidine and Nepsilon(carboxymethyl)lysine (CML)] in the uremic milieu. METHODS: Serum levels of the glycoxidation products, pentosidine and CML, were quantitated by high-performance liquid chromatography in uremic patients treated by dialysis. The formation of early glycation products (as furosine) and late glycoxidation products was modeled in uremic serum and in spent peritoneal dialysate. RESULTS: Clinical factors that affect circulating levels of AGEs included dialysis clearance and dialyzer membrane pore size, but not the presence or absence of diabetes. Both pentosidine and CML form at an accelerated rate in serum from uremic patients. Chelating agents most effectively slow the formation in vitro. In uremic fluids, the primary mechanism of formation of pentosidine is through the Amadori pathway. The primary mechanism of formation of CML is through metal-chelated autoxidation of reducing sugars generating reactive carbonyl precursors. In uremic serum, the presence of an unidentified reactive low molecular weight precursor accelerates the formation of pentosidine. CONCLUSIONS: The formation of the two glycoxidation products, pentosidine and CML, proceeds by different pathways and is enhanced by different precursors in the uremic milieu. The formation of both AGEs is markedly enhanced by metal catalyzed reactions, evidence for the presence of increased metal-ion mediated oxidant stress in uremia. PMID- 10844629 TI - Predicting acute renal failure after coronary bypass surgery: cross-validation of two risk-stratification algorithms. AB - BACKGROUND: Acute renal failure (ARF) requiring dialysis after coronary artery bypass grafting (CABG) occurs in 1 to 5% of patients and is independently associated with postoperative mortality, even after case-mix adjustment. A risk stratification algorithm that could reliably identify patients at increased risk of ARF could help improve outcomes. METHODS: To assess the validity and generalizability of a previously published preoperative renal risk-stratification algorithm, we analyzed data from the Quality Measurement and Management Initiative (QMMI)1 patient cohort. The QMMI includes all adult patients (N = 9498) who underwent CABG at 1 of 12 academic tertiary care hospitals from August 1993 to October 1995. ARF requiring dialysis was the outcome of interest. Cross validation of a recursive partitioning algorithm developed from the VA Continuous Improvement in Cardiac Surgery Program (CICSP) was performed on the QMMI. An additive severity score derived from logistic regression was also cross-validated on the QMMI. RESULTS: The CICSP recursive partitioning algorithm discriminated well (ARF vs. no ARF) in QMMI patients, even though the QMMI cohort was more diverse. Rates of ARF were similar among risk subgroups in the CICSP tree, as was the overall ranking of subgroups by risk. Using logistic regression, independent predictors of ARF in the QMMI cohort were similar to those found in the CICSP. The CICSP additive severity score performed well in the QMMI cohort, successfully stratifying patients into low-, medium-, high-, and very high-risk groups. CONCLUSIONS: The CICSP preoperative renal-risk algorithms are valid and generalizable across diverse populations. PMID- 10844628 TI - A role for leukotrienes in cyclosporine nephrotoxicity. AB - BACKGROUND: Nephrotoxicity associated with cyclosporine A (CsA) administration is characterized by marked renal vasoconstriction, interstitial fibrosis, and arteriolar hypertrophy. While the molecular mechanisms of CsA toxicity are not well characterized, previous studies have demonstrated that altered arachidonic acid (AA) metabolism plays a role its pathogenesis. Using a rat renal transplant model, the purpose of this study was to examine the effects of CsA on the 5 lipoxygenase (5-LO) pathway of AA metabolism. METHODS: The PVG (RT1c) strain of rats underwent kidney transplantation, and recipients of nonrejecting kidney transplants were treated with either 50 mg/kg/day CsA or vehicle (N = 24). To determine the physiologic significance of increased leukotriene (LT) production, the peptidoleukotriene receptor antagonist SKF 106203 was administered to CsA treated animals for six days. RESULTS: CsA caused a substantial reduction in glomerular filtration rate (GFR) in the transplanted rats compared with the vehicle-treated controls (1.5 +/- 0.6 vs. 4.1 +/- 0.8 mL/min/kg, P < 0.05). The reduction in renal function was associated with enhanced urinary excretion of the peptidoleukotriene metabolites LTE4 (1431 +/- 207 vs. 953 +/- 125 pg/24 h, P < 0.05) and N-acetyl-LTE4 (4411 +/- 848 vs. 463 +/- 70 pg/24 h, P < 0.001). LT receptor blockade had a significant protective effect on renal transplant function in CsA-treated animals (GFR, 4.8 +/- 1.1 vs. 1.7 +/- 0.9 mL/min/kg, P < 0.05), such that CsA-treated animals that received SKF106203 maintained GFR at levels similar to controls that never received CsA (4.1 +/- 0.8 mL/min/kg). Peptidoleukotriene receptor blockade also prevented the histomorphological abnormalities caused by CsA, including tubular vacuolization. CONCLUSIONS: These studies identify a critical role for LTs in the pathophysiology of CsA nephrotoxicity and suggest that LT antagonists may be useful in preventing CsA associated kidney toxicity. PMID- 10844630 TI - Tidal peritoneal dialysis: comparison of different tidal regimens and automated peritoneal dialysis. AB - BACKGROUND: The National Kidney Foundation Dialysis Outcomes Quality Initiative (DOQI) clinical practice guidelines have suggested minimal weekly Kt/V urea and creatinine clearance goals for peritoneal dialysis patients maintained on continuous ambulatory peritoneal dialysis (CAPD) and automated peritoneal dialysis (APD). Achieving these goals may present problems, particularly in larger patients whose residual renal function declines. Thus, modifications of the dialysis regimen, such as tidal peritoneal dialysis (TPD), have been developed. However, the ability of TPD to improve the efficiency of the dialysis procedure remains uncertain. METHODS: Stable, cycling peritoneal dialysis patients were placed into two groups to study the effectiveness of different TPD prescriptions on peritoneal clearances of urea and creatinine. The volume of dialysis solution used and the duration of therapy were fixed in the two groups. Comparisons were made to conventional APD using multiple hourly cycles in which spent dialysis solution was completely drained with each cycle. Group I patients received a total of 15 L of PD solution over 9.5 hours in the dialysis unit. These patients received 10, 25, and 50% TPD and APD on four separate days. Group II patients received 24 L of PD solution over 9.5 hours. These patients received 25 and 50% APD on separate days in the dialysis unit. Peritoneal dialysis clearances for urea (pKt/V) and creatinine (pCCr) levels were calculated for both groups. The results were then analyzed to determine whether there was any significant difference among the various prescriptions. RESULTS: The data in the group I patients indicated a mean daily pKt/V of 0.22 +/- 0.03 with 10% TPD, 0.23 +/- 0.02 with 25% TPD, 0.25 +/- 0.02 with 50% TPD, and 0.26 +/- 0.02 with APD. Paired t-test analysis for pKt/V demonstrated that 10 and 25% TPD resulted in significantly lower values than 50% TPD and APD (P < 0.05). Mean daily pCCr L/24 h/1.73 m2 was 6.03 +/- 0.72 for 10% TPD, 6.34 +/- 0.83 for 25% TPD, 6.65 +/- 0.51 for 50% TPD, and 7.01 +/- 0.96 for APD; these differences were not significantly different. The data in the group II patients demonstrated a mean daily pKt/V of 0.28 +/- 0.03 with 25% TPD, 0.29 +/- 0.05 with 50% TPD, and 0.30 +/- 0.05 for APD. The mean daily pCCr was 6.69 +/- 0.47 for 25% TPD, 8.09 +/- 1.30 for 50% TPD, and 7.63 +/- 1.13 for APD. There were no statistical differences for pKt/V and pCCr within the 24 L group. CONCLUSION: When the duration of therapy and volume of dialysate volume are kept constant, TPD does not result in an improvement in clearances compared with conventional APD, at least with dialysate volumes up to 24 L. PMID- 10844631 TI - In vivo visualization of hemodialysis-induced alterations in leukocyte endothelial interactions. AB - BACKGROUND: The aim of this study was to develop a model for hemodialysis (HD) in small animals using conventional dialysis equipment that would allow the intravital microscopic observation of leukocyte-endothelial interactions in vivo. METHODS: Cuprophan dialyzers were adapted to obtain a similar ratio of membrane area to blood volume as in clinical HD. A silicone ring was inserted into the dialyzer's inlet to limit the number of blood-perfused capillaries. Rabbits were dialyzed for one hour without a dialysate flow. RESULTS: Extracorporeal circulation with the cuprophan dialyzer resulted in a transient leukopenia and complement activation. At the nadir of leukopenia, leukocytes that rolled along the venular wall were scarcely observed, whereas rolling was abundant (54 +/- 9 per min) prior to extracorporeal circulation. The adhesion of leukocytes to the vascular endothelium was not induced. After 60 minutes, rolling of leukocytes was still reduced by 73 +/- 5.5%, despite the full recovery of circulating leukocyte counts. Extracorporeal circulation without a dialyzer also tended to reduce leukocyte rolling, although systemic leukocyte counts were not affected. CONCLUSIONS: The use of adapted conventional cuprophan hemodialyzers in rabbits yielded a transient leukopenia similar to that in clinical HD. Using intravital microscopy, we demonstrated impairment of leukocyte-endothelial interactions. In addition, our data indicate that tissues, in which leukocytes can roll and adhere, are not automatically sites of leukocyte sequestration during HD-induced leukopenia. PMID- 10844632 TI - Renal allograft protection with losartan in Fisher-->Lewis rats: hemodynamics, macrophages, and cytokines. AB - BACKGROUND: We sought to assess the effects of angiotensin receptor blockade on glomerular hypertension, macrophage recruitment, and cytokine expression, all of which contribute to the development of chronic graft injury in this model. METHODS: The effects of treatment with the specific angiotensin II type 1 (AT1) receptor antagonist, losartan, were assessed over 24 weeks in F344-->LEW rats (LOS, N = 9) versus vehicle-treated F344-->LEW controls (CON, N = 9). RESULTS: UprotV rose progressively in CON (from 7.0 +/- 2.9 to 41 +/- 17 mg/day at 24 wk) but remained at baseline in LOS (4.2 +/- 0.6 to 9.4 +/- 1.3 mg/day, P < 0.05 vs. CON). Glomerular capillary pressure (PGC) was increased in CON (71 +/- 1 mm Hg at week 20), but remained within the normal range in LOS rats (54 +/- 2 mm Hg, P < 0.05). Glomerulosclerosis averaged 0.3 +/- 0.2% in LOS versus 4 +/- 2% in CON rats (P < 0.05). Tubulointerstitial injury was minimal in both LOS and CON rats (+). The overexpression of renal cortical cytokine mRNA levels for the monocyte chemoattractants, monocyte chemoattractant protein-1 (MCP-1) and RANTES, as well as interleukin-1, inducible nitric oxide synthase, and transforming growth factor beta, assessed by competitive reverse transcription-polymerase chain reaction, was suppressed in LOS versus CON rats at 20 weeks. Macrophage and T-cell numbers were decreased, and MCP-1, RANTES, and intercellular adhesion molecule-1 staining in the graft, identified by immunohistochemistry, were attenuated in LOS versus CON rats. CONCLUSIONS: The renoprotective effects of losartan in F344-->LEW rats were associated with lowered PGC, inhibition of macrophage chemoattractants and recruitment, and suppression of macrophage-associated cytokines at 20 weeks. These findings suggest that chronic allograft injury in F344-->LEW rats is, to a large extent, mediated by angiotensin II-dependent mechanisms and that these involve glomerular hemodynamics, macrophages, and macrophage-associated cytokines. PMID- 10844633 TI - Pentoxifylline inhibits human peritoneal mesothelial cell growth and collagen synthesis: effects on TGF-beta. AB - BACKGROUND: Prevention or treatment of peritoneal fibrosing syndrome has become an important issue in patients on continuous ambulatory peritoneal dialysis (CAPD). Recent evidence has suggested that mesothelial stem cell proliferation and matrix over-production predispose the development of peritoneal fibrosis. We investigated whether pentoxifylline (PTX) affects human peritoneal mesothelial cell (HPMC) growth and collagen synthesis. METHODS: HPMC was cultured from human omentum by an enzymic disaggregation method. Cell proliferation was assayed using a methyltetrazolium uptake method. Cell cycle analysis was performed by flow cytometry. Collagen synthesis was measured by 3H-proline incorporation into pepsin-resistant, salt-precipitated collagen. Prostaglandins and cAMP were determined by enzyme immunoassay. Northern blot analysis was used to determine mRNA expression. RESULTS: Our data show that PTX inhibited serum-stimulated HPMC growth and collagen synthesis in a dose-dependent manner. Cell cycle analysis showed that PTX arrested the HPMCs in the G1 phase. PTX decreased the procollagen alpha1 (I) mRNA expression either stimulated by serum or transforming growth factor-beta (TGF-beta). PTX did not alter prostaglandins synthesis but dose dependently increased intracellular cAMP level. PTX, the same as 3-isobutyl-l methylxanthine, could potentiate prostaglandin E1 (PGE1) increased cAMP levels of HPMC. The antimitogenic and antifibrogenic effects of PTX on HPMC were reversed by N-[2]-((p-Bromocinnamyl)amino)ethyl]-5-isoquinolinesulfonamide (H-89). Therefore, the mechanism of these effects may be due to the phospodiesterase inhibitory property of PTX. CONCLUSIONS: These data suggest that PTX may have a role in treating peritoneal fibrosing syndrome. PMID- 10844634 TI - Analysis of the effect of hemodialysis on peripheral and central arterial pressure waveforms. AB - BACKGROUND: Arterial stiffening is very pronounced in renal patients. Carotid artery stiffening is a powerful predictor of future cardiovascular mortality, and measures of arterial compliance correlate much better with left ventricular mass (LVM) in dialysis patients than does brachial artery blood pressure (BP). The aim of our study was to describe the influence of a hemodialysis (HD) session on arterial cushioning function and to correlate the potential different types of behavior with echocardiographic derived parameters. METHODS: Radial artery pressure waveforms were measured and recorded noninvasively by applanation tonometry in 51 healthy patients on regular three times weekly HD. The data were then converted into aortic pressure waveforms using a regression equation (SphymoCortrade mark apparatus). Measurements were done pre- and post-HD in order to ascertain the effect of a single HD session on arterial hemodynamics. The augmentation index (AGI; the difference between early and late pressure peaks divided by the pulse pressure amplitude) was used as an index for vascular compliance. Reproducibility was assessed in 20 young healthy subjects by determining the aortic pulse wave augmentation index twice from radial artery BP measurements one minute apart. Intraobserver error was 2.4%. For 10 dialysis patients similarly studied, the intraobserver error was 1.6%. RESULTS: AGI was correlated with subjects' height (r = -0.37, P = 0.009), weight (r = -0.41, P = 0.002), and BP levels: radial systolic BP (r = 0.33, P = 0.018), radial diastolic BP (r = 0.29, P = 0.036), and central systolic BP (r = 0.51, P < 0.001). Comparing the pre- with the post-HD AGI values, four patterns of evolution became apparent: (1) The AGI was negative before the HD session and became even more negative afterward (N = 3 out of 51). (2) The AGI was positive before the HD session but became negative after dialysis (N = 19 out of 51). (3) The AGI was positive before the HD session and, although diminished afterward, remained positive (N = 23 out of 51). (4) The AGI was positive before the HD session and increased afterward (N = 6 out of 51). We also found that in some patients, AGI remained at lower than predialysis levels for at least 24 hours. Significant relationships between echocardiographic parameters and pulse wave contour (PWC) variables included pre-HD AGI and LVM (r = 0.47, P < 0. 001). There was better correlation between LVM and derived predialysis aortic systolic BP (r = 0.56, P < 0.001) than measured brachial (peripheral) systolic BP (r = 0.35, P = 0.04). Patients whose waveform remained abnormal (AGI remained positive) after HD had a more dilated LV (LV-EDD = 52.07 +/- 3.48 mm) than did those patients for whom HD restored "normal" arterial hemodynamics (LV-EDD 46.86 +/- 4.06 mm, P < 0.05). CONCLUSIONS: A standard HD session profoundly affected aortic BP waveform characteristics, with a reduction in wave reflection in 88% of patients. However, restoration by HD of a normal aortic waveform was unusual. Patients whose waveform remained abnormal after HD had larger more dilated LV chambers than did those patients for whom HD restored "normal" arterial hemodynamics. PMID- 10844635 TI - Methods for estimating the volume of individual glomeruli. AB - Methods for estimating the volume of individual glomeruli. BACKGROUND: The Cavalieri and maximal planar area (MPA) methods are commonly used to measure the volume of individual glomeruli. Previous studies have suggested that the MPA method, which is less laborious, yields values that are much greater than those obtained by the Cavalieri method. The current study re-examined the relationship of MPA and Cavalieri values for glomerular volume in humans and rats. METHODS: Both methods were used to measure the volume of 1201 glomeruli from 58 humans and 281 glomeruli from 15 rats. Tissue was embedded in Epon. Further mathematical analysis was performed to assess the extent to which deviation of glomeruli from spherical shape affects the relationship of values obtained by the MPA and Cavalieri methods. RESULTS: MPA values exceeded Cavalieri values by an average of only 14 +/- 22% in humans and 6 +/- 16% in rats. The relationship of MPA to Cavalieri values was similar in individual humans and rats, with widely varying values for average glomerular volume. Neither the development of sclerosis nor the loss of any connection to a tubule affected the relationship of the MPA and Cavalieri values for the volume of individual glomeruli. Mathematical analysis showed that MPA values would not exceed Cavalieri values if glomeruli had ellipsoidal rather than spherical shape. CONCLUSION: Similar values for glomerular volume are obtained using the Cavalieri and MPA methods in humans and rats. PMID- 10844636 TI - Many cell types are Shiga toxin targets. PMID- 10844637 TI - Tight junctions: guardians of the paracellular pathway. PMID- 10844638 TI - Serum magnesium and parathyroid hormone levels in dialysis patients. PMID- 10844639 TI - Home hemodialysis: excellent survival at less cost, but still underutilized. PMID- 10844640 TI - Blood pressure and prediction of patient outcome. PMID- 10844641 TI - Lowering C-peptide levels and renoprotective therapy. PMID- 10844642 TI - New tools in diagnosing renal artery stenosis. PMID- 10844643 TI - PriA and phage T4 gp59: factors that promote DNA replication on forked DNA substrates microreview. AB - The initiation of DNA synthesis on forked DNA templates is a vital process in the replication and maintenance of cellular chromosomes. Two proteins that promote replisome assembly on DNA forks have so far been identified. In phage T4 development the gene 59 protein (gp59) assembles replisomes at D-loops, the sites of homologous strand exchange. Bacterial PriA protein plays an analogous function, most probably restarting replication after replication fork arrest with the aid of homologous recombination proteins, and PriA is also required for phage Mu replication by transposition. Gp59 and PriA exhibit similar DNA fork binding activities, but PriA also has a 3' to 5' helicase activity that can promote duplex opening for replisome assembly. The helicase activity allows PriA's repertoire of templates to be more diverse than that of gp59. It may give PriA the versatility to restart DNA replication without recombination on arrested replication forks that lack appropriate duplex openings. PMID- 10844644 TI - Mutational analysis of the functional motifs of RuvB, an AAA+ class helicase and motor protein for holliday junction branch migration. AB - Escherichia coli RuvB protein, together with RuvA, promotes branch migration of Holliday junctions during homologous recombination and recombination repair. The RuvB molecular motor is an intrinsic ATP-dependent DNA helicase with a hexameric ring structure and its architecture has been suggested to be related to those of the members of the AAA+ protein class. In this study, we isolated a large number of plasmids carrying ruvB mutant genes and identified amino acid residues important for the RuvB functions by examining the in vivo DNA repair activities of the mutant proteins. Based on these mutational studies and amino acid conservation among various RuvBs, we identified 10 RuvB motifs that agreed well with the features of the AAA+ protein class and that distinguished the primary structure of RuvB from that of typical DNA/RNA helicases with seven conserved helicase motifs. PMID- 10844645 TI - Biosynthesis of carbapenem antibiotic and prodigiosin pigment in Serratia is under quorum sensing control. AB - Serratia sp. ATCC 39006 produces the carbapenem antibiotic, carbapen-2-em-3 carboxylic acid and the red pigment, prodigiosin. We have previously reported the characterization of a gene, carR, controlling production of carbapenem in this strain. We now describe further characterization of the carR locus to locate the genes encoding carbapenem biosynthetic and resistance functions. A novel family of diverse proteins showing sequence similarity to the C-terminal domain of CarF (required for carbapenem resistance) is described. We also report the isolation of the locus involved in the biosynthesis of the red pigment, prodigiosin. A cosmid containing approximately 35 kb of the Serratia chromosome encodes synthesis of the pigment in the heterologous host, Erwinia carotovora, demonstrating, for the first time, that the complete prodigiosin biosynthetic gene cluster had been cloned and functionally expressed. We report the isolation of a third locus in Serratia, containing convergently transcribed genes, smaI and smaR, encoding LuxI and LuxR homologues respectively. SmaI directs the synthesis of N-acyl homoserine lactones involved in the quorum sensing process. We demonstrate that biosynthesis of the two secondary metabolites, carbapenem antibiotic and prodigiosin pigment, is under pheromone-mediated transcriptional regulation in this bacterium. Finally, we describe a new prodigiosin-based bioassay for detection of some N-acyl homoserine lactones. PMID- 10844646 TI - Analysis of the DNA-binding domain of Escherichia coli DnaA protein. AB - The DNA-binding domain of the Escherichia coli DnaA protein is represented by the 94 C-terminal amino acids (domain 4, aa 374-467). The isolated DNA-binding domain acts as a functional repressor in vivo, as monitored with a mioC:lacZ translational fusion integrated into the chromosome of the indicator strain. In order to identify residues required for specific DNA binding, site-directed and random PCR mutagenesis were performed, using the mioC:lacZ construct for selection. Mutations defective in DNA binding were found all over the DNA-binding domain with some clustering in the basic loop region, within presumptive helix B and in a highly conserved region at the N-terminus of presumptive helix C. Surface plasmon resonance (SPR) analysis revealed different binding classes of mutant proteins. No or severely reduced binding activity was demonstrated for amino acid substitutions at positions R399, R407, Q408, H434, T435, T436 and A440. Altered binding specificity was found for mutations in a 12 residue region close to the N-terminus of helix C. The defects of the classical temperature sensitive mutants dnaA204, dnaA205 and dnaA211 result from instability of the proteins at higher temperatures. dnaX suppressors dnaA71 and dnaA721 map to the region close to helix C and bind DNA non-specifically. PMID- 10844647 TI - Enzyme I and HPr from Lactobacillus casei: their role in sugar transport, carbon catabolite repression and inducer exclusion. AB - We have cloned and sequenced the Lactobacillus casei ptsH and ptsI genes, which encode enzyme I and HPr, respectively, the general components of the phosphoenolpyruvate-carbohydrate phosphotransferase system (PTS). Northern blot analysis revealed that these two genes are organized in a single-transcriptional unit whose expression is partially induced. The PTS plays an important role in sugar transport in L. casei, as was confirmed by constructing enzyme I-deficient L. casei mutants, which were unable to ferment a large number of carbohydrates (fructose, mannose, mannitol, sorbose, sorbitol, amygdaline, arbutine, salicine, cellobiose, lactose, tagatose, trehalose and turanose). Phosphorylation of HPr at Ser-46 is assumed to be important for the regulation of sugar metabolism in Gram positive bacteria. L. casei ptsH mutants were constructed in which phosphorylation of HPr at Ser-46 was either prevented or diminished (replacement of Ser-46 of HPr with Ala or Thr respectively). In a third mutant, Ile-47 of HPr was replaced with a threonine, which was assumed to reduce the affinity of P-Ser HPr for its target protein CcpA. The ptsH mutants exhibited a less pronounced lag phase during diauxic growth in a mixture of glucose and lactose, two PTS sugars, and diauxie was abolished when cells were cultured in a mixture of glucose and the non-PTS sugars ribose or maltose. The ptsH mutants synthesizing Ser-46-Ala or Ile-47-Thr mutant HPr were partly or completely relieved from carbon catabolite repression (CCR), suggesting that the P-Ser-HPr/CcpA-mediated mechanism of CCR is common to most low G+C Gram-positive bacteria. In addition, in the three constructed ptsH mutants, glucose had lost its inhibitory effect on maltose transport, providing for the first time in vivo evidence that P-Ser-HPr participates also in inducer exclusion. PMID- 10844648 TI - Phase variation of the gonococcal siderophore receptor FetA. AB - FetA, the recently characterized gonococcal ferric enterobactin receptor, exhibited extremely rapid phase variation between high- and low-expression levels. The frequency of phase variation was approximately 1.3% in both directions in gonococcal strain FA1090. FetA expression in the 'high phase' was significantly greater than the level of expression in the 'low phase'. Expression levels correlated with the number of cytosine residues in a string of cytosines located close to the transcriptional start site for fetA between the putative -10 and -35 consensus sequences. Antibody production against FetA commonly occurs in infected patients, and we therefore hypothesize that phase variation reflects a balance between the advantages of being able to use a ferric siderophore as an iron source and evasion of the host immune response. PMID- 10844649 TI - Control of bioluminescence in Vibrio fischeri by the LuxO signal response regulator. AB - Bioluminescence in the marine bacterium Vibrio fischeri is controlled by the excretion of a N-acyl homoserine lactone (HSL) autoinducer which interacts with a regulator, LuxR, and activates transcription of the lux operon at high-cell density. This system has become the prototype for quorum sensing in many bacteria. Although light emission in Vibrio harveyi is also regulated by a N-acyl HSL inducer, in sharp contrast, a completely different and more complex system is involved in quorum sensing which is mediated via LuxO, the response regulator of a phosphorelay signal transduction system. In the present work, luxO and the overlapping luxU gene, also involved in the phosphorelay system in V. harveyi, have been discovered in V. fischeri. By gene replacement technology, a V. fischeri luxO- mutant was generated whose phenotype was similar to that of V. harveyi luxO- showing that LuxO is involved in control of luminescence in V. fischeri. This mutant could be complemented with luxO from either V. fischeri or V. harveyi resulting in the restoration of the dependence of luminescence intensity on cell density. In contrast to V. harveyi luxO-, light emission of V. fischeri luxO- was stimulated by the N-acyl-HSL autoinducer indicating that luxO is part of a second signal transduction system controlling luminescence in this species. The presence of a luxO-based phosphorelay regulatory system as well as the luxR-based system in V. fischeri suggests that the former system, originally discovered in V. harveyi, may be a general regulatory mechanism in luminescent bacteria. PMID- 10844650 TI - Subcellular localization of the Agrobacterium tumefaciens T-DNA transport pore proteins: VirB8 is essential for the assembly of the transport pore. AB - Agrobacterium tumefaciens transforms plants by transferring DNA to the plant cell nucleus. The VirB membrane proteins are postulated to form a pore for the transport of the DNA across the bacterial membranes. Immunofluorescence and immunoelectron microscopy were used to study the transport pore complex. Three likely components of the transport pore, VirB8, VirB9 and VirB10, localized primarily to the inner membrane, outer membrane and periplasm respectively. A significant amount of VirB10 was also found associated with the outer membrane. When expressed alone VirB9 and VirB10 were randomly distributed along the cell membrane. Subcellular location of both proteins changed dramatically in the presence of the other VirB proteins. Both proteins localized to fewer sites and most of the gold particles representing protein molecules were found in clusters suggesting that the two proteins are in a protein complex. VirB8, on the other hand, localized to clusters even in the absence of the other VirB proteins. To investigate the role of VirB8 in the formation of VirB9 and VirB10 protein complexes, we studied the effect of deletion of virB8 on the subcellular location of VirB9 and VirB10. In a virB8 deletion mutant both proteins were distributed randomly on the cell membrane indicating that VirB8 is essential for complex assembly. PMID- 10844651 TI - Analysis of the oxidative stress regulation of the Candida albicans transcription factor, Cap1p. AB - CAP1 encodes a basic region-leucine zipper (bZip) transcriptional regulatory protein that is required for oxidative stress tolerance in Candida albicans. Cap1p is a homologue of a Saccharomyces cerevisiae bZip transcription factor designated Yap1p that is both required for oxidative stress tolerance and localized to the nucleus in response to the presence of oxidants. Oxidant regulated localization of Yap1p to the nucleus requires the presence of a carboxy terminal cysteine residue (C629) that is conserved in Cap1p as C477. To examine the role of this conserved cysteine residue, C477 was replaced with an alanine residue. This mutant protein, C477A Cap1p, was analysed for its behaviour both in S. cerevisiae and C. albicans. Wild type and C477A Cap1p were able to complement the oxidant hypersensitivity of a Deltayap1 S. cerevisiae strain. Whereas a Yap1p responsive lacZ fusion gene was oxidant inducible in the presence of YAP1, the C. albicans Cap1p derivatives were not oxidant responsive in S. cerevisiae. Introduction of wild type and C477A Cap1p-expressing plasmids into C. albicans produced differential resistance to oxidants. Glutathione reductase activity was found to be inducible by oxidants in the presence of Cap1p but was constitutively elevated in the presence of C477A Cap1p. Western blot assays indicate Cap1p is post-translationally regulated by oxidants. Green fluorescent protein fusions to CAP1 showed that this protein is localized to the nucleus only in the presence of oxidants while C477A Cap1p is constitutively nuclear localized. Directly analogous to S. cerevisiae Yap1p, regulated nuclear localization of C. albicans Cap1p is crucial for its normal function. PMID- 10844652 TI - Oxygenated mycolic acids are necessary for virulence of Mycobacterium tuberculosis in mice. AB - Members of the Mycobacterium tuberculosis group synthesize a family of long-chain fatty acids, mycolic acids, which are located in the cell envelope. These include the non-oxygenated alpha-mycolic acid and the oxygenated keto- and methoxymycolic acids. The function in bacterial virulence, if any, of these various types of mycolic acids is unknown. We have constructed a mutant strain of M. tuberculosis with an inactivated hma (cmaA, mma4) gene; this mutant strain no longer synthesizes oxygenated mycolic acids, has profound alterations in its envelope permeability and is attenuated in mice. PMID- 10844653 TI - Genes required for cytochrome c synthesis in Bacillus subtilis. AB - Cytochromes of c-type contain covalently bound haem and in bacteria are located on the periplasmic side of the cytoplasmic membrane. More than eight different gene products have been identified as being specifically required for the synthesis of cytochromes c in Gram-negative bacteria. Corresponding genes are not found in the genome sequences of Gram-positive bacteria. Using two random mutagenesis approaches, we have searched for cytochrome c biogenesis genes in the Gram-positive bacterium Bacillus subtilis. Three genes, resB, resC and ccdA, were identified. CcdA has been found previously and is required for a late step in cytochrome c synthesis and also plays a role in spore synthesis. No function has previously been assigned for ResB and ResC but these predicted membrane proteins show sequence similarity to proteins required for cytochrome c synthesis in chloroplasts. Attempts to inactivate resB and resC in B. subtilis have indicated that these genes are essential for growth. We demonstrate that various nonsense mutations in resB or resC can block synthesis of cytochromes c with no effect on other types of cytochromes and little effect on sporulation and growth. The results strongly support the recent proposal that Gram-positive bacteria, cyanobacteria, epsilon-proteobacteria, and chloroplasts have a similar type of machinery for cytochrome c synthesis (System II), which is very different from those of most Gram-negative bacteria (System I) and mitochondria (System III). PMID- 10844654 TI - Expression of yeast INM1 encoding inositol monophosphatase is regulated by inositol, carbon source and growth stage and is decreased by lithium and valproate. AB - Inositol monophosphatase plays a vital role in the de novo biosynthesis of inositol and in the phosphoinositide second messenger signalling pathway. We cloned the Saccharomyces cerevisiae open reading frame (ORF) YHR046c (termed INM1), which encodes inositol monophosphatase, characterized the protein Inm1p and analysed expression of the INM1 gene. INM1 was expressed in bacteria under the control of the lacZ promoter. The purified protein has inositol monophosphatase activity that is inhibited by the antibipolar drug lithium, but not valproate. In the inm1Delta:URA3 null mutant, inositol monophosphatase activity was reduced but not eliminated. The disruption had little effect on growth in the presence of lithium or valproate and no effect on growth in the absence of inositol. To characterize the regulation of INM1, we examined the effects of inositol, carbon source, growth phase, and the antibipolar drugs lithium and valproate on INM1 expression using an INM1-lacZ reporter gene. Unlike all other phospholipid biosynthetic enzyme-encoding genes studied, which contain the UASINO regulatory element, INM1 expression is increased in the presence of inositol. In addition, INM1 expression was repressed during growth in glycerol and derepressed as glucose-grown cells entered stationary. Both lithium and valproate, which cause a decrease in intracellular inositol, effect a decrease in INM1 expression. A model is presented to account for regulation of INM1 expression. PMID- 10844655 TI - AglU, a protein required for gliding motility and spore maturation of Myxococcus xanthus, is related to WD-repeat proteins. AB - The aglU gene of Myxococcus xanthus encodes a protein similar to Het-E1 (vegetative incompatibility) from Podospora anserina, acylaminoacyl-peptidase from Bacillus subtilis, and TolB from Escherichia coli. These proteins all have evenly spaced SPDG repeats that are characteristic of a larger motif called the WD-repeat. The WD-repeat is predicted to form a beta-propeller structure that mediates the assembly of heteromeric protein complexes. AglU has a consensus lipoprotein attachment motif that includes a type II signal sequence followed by a cysteine residue. This suggests that AglU is matured, then attached to the outer membrane via fatty acid acylation at this Cys. Cells carrying a mutation in aglU are blocked in adventurous gliding and can swarm only if cells are in contact with one another. When starved of nutrients, the aglU mutant aggregates and forms multicellular fruiting bodies like the wild-type strain, but is unable to produce heat-resistant spores. This suggests that adventurous gliding motility, per se, is not required for development, but that AglU is essential for a terminal step of spore differentiation. PMID- 10844656 TI - Identification of genes affecting selenite toxicity and resistance in Saccharomyces cerevisiae. AB - Recent studies associating dietary selenium with reduced cancer susceptibility have aroused interest in this substance. In the millimolar range, selenite is toxic and slightly mutagenic for yeast. We show that selenite-treated yeast cells tend to arrest as large budded cells and that this arrest is abolished in a rad9 mutant that is significantly sensitive to selenite. Interestingly, a rev3 mutant affected in the error-prone repair pathway is also sensitive to selenite, whereas mutations in the other DNA repair pathways do not strongly affect resistance to selenite. We propose that selenite treatment leads to DNA damage inducing the RAD9-dependent cell cycle arrest. Selenite-induced DNA damage could be converted to mutations by the Rev3p-dependent lesion bypass system, thus allowing the cell cycle to progress. We have also investigated the selenite detoxification mechanisms and identified three genes involved in this process. In the present study, we show that lack of the cadmium glutathione-conjugate vacuolar pump Ycf1p or overexpression of the sulphite resistance membrane protein Ssu1p enhance the capacity of yeast cells to resist selenite treatment. Finally, we show that overexpression of the glutathione reductase Glr1p increases resistance to selenite, suggesting that selenite toxicity in yeast is closely linked to its oxidative capacity. PMID- 10844657 TI - Competence regulation by oxygen in Streptococcus pneumoniae: involvement of ciaRH and comCDE. AB - Anaerobic aerotolerant Streptococcus pneumoniae modulates its genetic transformability and its virulence in response to the oxygen concentration. The activity of a single protein encoded by nox and showing NADH oxidase activity is involved in these adaptive responses to O2. Northern blot analysis of wild-type cultures grown under aerobic and microaerobic conditions indicated transcriptional control of comCDE by O2. An O2-independent mutant strain carrying the gain-of-function mutation comE38KE was isolated and its analysis showed that ComE is a key point in competence stimulation by O2. Plasmid insertion mutations in ciaRH revealed that this two component signal-transducing system negatively regulates comCDE transcription. The level of comCDE transcripts appears as a major control point in competence regulation by O2 and also by growth phase and cell density. PMID- 10844658 TI - The 5'-proximal hairpin loop of lbi RNA is a key structural element in repression of D-galactan II biosynthesis in Klebsiella pneumoniae serotype O1. AB - The lbi (lipopolysaccharide biosynthesis interfering) RNA of phage Acm1, an untranslated RNA transcript of 97 nucleotides, previously shown to affect O polysaccharide biosynthesis in various Escherichia coli strains, was found to downregulate the synthesis of the D-galactan II component of the O-specific polysaccharide in Klebsiella pneumoniae serotype O1. Enzymatic and Pb2+ probing experiments revealed that lbi RNA consists of two consecutive stem-loop structures, the 5'-proximal hairpin loop of 15 nucleotides being particularly accessible to single strand-specific probes. Based on the assumption that the 5' proximal hairpin loop may be involved in an antisense interaction with cellular target RNAs, we randomly mutagenized one or two of its central nucleotides. Expression of mutated lbi RNA variants in K. pneumoniae serotype O1 relieved at least partly the repression of D-galactan II formation. In addition, a truncated version of lbi RNA lacking the 3'-proximal hairpin loop was almost as efficient as the wild-type RNA in downregulating D-galactan II synthesis. The results obtained indicate that the 5'-proximal hairpin loop of lbi RNA functions as a key structural element in the mechanism leading to the inhibition of D-galactan II biosynthesis in K. pneumoniae serotype O1. PMID- 10844659 TI - Comparative proteome analysis of Helicobacter pylori. AB - Helicobacter pylori, the causative agent of gastritis, ulcer and stomach carcinoma, infects approximately half of the worlds population. After sequencing the complete genome of two strains, 26695 and J99, we have approached the demanding task of investigating the functional part of the genetic information containing macromolecules, the proteome. The proteins of three strains of H. pylori, 26695 and J99, and a prominent strain used in animal models SS1, were separated by a high-resolution two-dimensional electrophoresis technique with a resolution power of 5000 protein spots. Up to 1800 protein species were separated from H. pylori which had been cultivated for 5 days on agar plates. Using matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS) peptide mass fingerprinting we have identified 152 proteins, including nine known virulence factors and 28 antigens. The three strains investigated had only a few protein spots in common. We observe that proteins with an amino acid exchange resulting in a net change of only one charge are shifted in the two-dimensional electrophoresis (2-DE) pattern. The expression of 27 predicted conserved hypothetical open reading frames (ORFs) and six unknown ORFs were confirmed. The growth conditions of the bacteria were shown to have an effect on the presence of certain proteins. A preliminary immunoblotting study using human sera revealed that this approach is ideal for identifying proteins of diagnostic or therapeutic value. H. pylori 2-DE patterns with their identified protein species were added to the dynamic 2D-PAGE database (http://www.mpiib-berlin.mpg.de/2D-PAGE/). This basic knowledge of the proteome in the public domain will be an effective instrument for the identification of new virulence or pathogenic factors, and antigens of potentially diagnostic or curative value against H. pylori. PMID- 10844660 TI - Species-specific binding of human secretory component to SpsA protein of Streptococcus pneumoniae via a hexapeptide motif. AB - SpsA, a pneumococcal surface protein belonging to the family of choline-binding proteins, interacts specifically with secretory immunglobulin A (SIgA) via the secretory component (SC). SIgA and free SC from mouse, rat, rabbit and guinea-pig failed to interact with SpsA indicating species-specific binding to human SIgA and SC. SpsA is the only pneumococcal receptor molecule for SIgA and SC as confirmed by complete loss of SIgA and SC binding to a spsA mutant. Analysis of recombinant SpsA fusion proteins showed that the binding domain is located in the N-terminal region of SpsA. By the use of different truncated N-terminal SpsA fusion proteins, the minimum binding domain was shown to be composed of 112 amino acids (residues 172-283). The sequence of this 112-amino-acids domain was used to spot synthesize 34 overlapping peptides, consisting of 15 amino acids each, with an offset of three amino acids on a cellulose membrane. One of the peptides reacted specifically with both SIgA and SC. By using a second membrane with immobilized synthetic peptides of decreasing length containing parts of the identified 15-amino-acid motif a hexapeptide, YRNYPT was identified as the binding motif for SC and SIgA. SpsA proteins with a size smaller than the assay positive domain of 112 amino acids were able to inhibit the interaction of SIgA and pneumococci provided they contained the binding motif. The results indicated that the hexapeptide YRNYPT located in SpsA of pneumococcal strain type 1 (ATCC 33400) between amino acids 198 and 203 is involved in SIgA and SC binding. Because synthetic peptides containing only parts of the hexapeptide also assayed positive, these results further suggest that at least the amino acids YPT of the identified hexapeptide are critical for binding to SC and SIgA. Amino acid substitutions in the identified putative binding motif abolished SC-/SIgA-binding activity of the mutated SpsA protein, confirming the functional activity of this hexapeptide and the critical role of the amino acids YPT in SC and SIgA binding. Identification of this motif, which is highly conserved in SpsA protein among different serotypes, might contribute towards a new peptide based vaccine strategy. PMID- 10844661 TI - GAP activity of the Yersinia YopE cytotoxin specifically targets the Rho pathway: a mechanism for disruption of actin microfilament structure. AB - The YopE cytotoxin of Yersinia pseudotuberculosis is an essential virulence determinant that is injected into the eukaryotic target cell via a plasmid encoded type III secretion system. Injection of YopE into eukaryotic cells induces depolymerization of actin stress fibres. Here, we show that YopE exhibits a GTPase-activating protein (GAP) activity and that the presence of YopE stimulates downregulation of Rho, Rac and Cdc42 activity. YopE has an arginine finger motif showing homology with those found in other GAP proteins. Exchange of arginine 144 with alanine, located in this arginine finger motif, results in an inactive form of YopE that can no longer stimulate GTP hydrolysis by the GTPase. Furthermore, a yopE(R144A) mutant is unable to induce cytotoxicity on cultured HeLa cells in contrast to the corresponding wild-type strain. Expression of wild type YopE in cells of Saccharomyces cerevisiae inhibits growth, while in contrast, expression of the inactive form of YopE, YopE(R144A), does not affect the yeast cells. Co-expression of proteins belonging to the Rho1 pathway of yeast, Rho1, Rom2p, Bck1 and Ste20, suppressed the growth phenotype of YopE in yeast cells. These results provide evidence that YopE exhibits a GAP activity to inactivate RhoGTPases, leading to depolymerization of the actin stress fibres in eukaryotic cells and growth inhibition in yeast. PMID- 10844662 TI - Salmonella SsrB activates a global regulon of horizontally acquired genes. AB - Salmonella enterica is a bacterial pathogen of humans that can proliferate within epithelial cells as well as professional phagocytes of the immune system. This ability requires an S. enterica specific locus termed Salmonella pathogenicity island 2 (SPI-2). SPI-2 encodes a type III secretion system that injects effectors encoded within the island into host cell cytosol to promote virulence. SsrAB is a two-component regulator encoded within SPI-2 that was assumed to activate SPI-2 genes exclusively. Here, it is shown that SsrB in fact activates a global regulon. At least 10 genes outside SPI-2 are SsrB regulated within epithelial and macrophage cells. Nine of these 10 SsrB-regulated genes outside SPI-2 reside within previously undescribed regions of the Salmonella genome. Most share no sequence homology with current database entries. However, one is remarkably homologous to human glucosyl ceramidase, an enzyme involved in the ceramide signalling pathway. The SsrB regulon is modulated by the two-component regulatory systems PhoP/PhoQ and OmpR/EnvZ, and is upregulated in the intracellular microenvironment. PMID- 10844663 TI - Variable human minisatellite-like regions in the Mycobacterium tuberculosis genome. AB - Mycobacterial interspersed repetitive units (MIRUs) are 40-100 bp DNA elements often found as tandem repeats and dispersed in intergenic regions of the Mycobacterium tuberculosis complex genomes. The M. tuberculosis H37Rv chromosome contains 41 MIRU loci. After polymerase chain reaction (PCR) and sequence analyses of these loci in 31 M. tuberculosis complex strains, 12 of them were found to display variations in tandem repeat copy numbers and, in most cases, sequence variations between repeat units as well. These features are reminiscent of those of certain human variable minisatellites. Of the 12 variable loci, only one was found to vary among genealogically distant BCG substrains, suggesting that these interspersed bacterial minisatellite-like structures evolve slowly in mycobacterial populations. PMID- 10844664 TI - Antibiotic resistance: era of the multidrug pump. PMID- 10844665 TI - N-terminal conservation of putative type III secreted effectors of Salmonella typhimurium. PMID- 10844666 TI - New functions for the ancient globin family: bacterial responses to nitric oxide and nitrosative stress. AB - Globin-like oxygen-binding proteins occur in bacteria, yeasts and other fungi, and protozoa. The simplest contain protohaem as sole prosthetic group, but show considerable variation in their similarity to the classical animal globins and plant globins. Flavohaemoglobins comprise a haem domain homologous to classical globins and a ferredoxin-NADP+ reductase (FNR)-like domain that converts the globin into an NAD(P)H-oxidizing protein with diverse reductase activities. In Escherichia coli, the prototype flavohaemoglobin (Hmp) is clearly involved in responses to nitric oxide (NO) and nitrosative stress: (i) the structural gene hmp is upregulated by NO and nitrosating agents; (ii) purified Hmp binds NO avidly, but also converts it to nitrate (aerobically) or nitrous oxide (anaerobically); (iii) hmp mutants are hypersensitive to NO and nitrosative stresses. Here, we review recent advances in E. coli and the growing number of microbes in which globins are known, draw particular attention to the essential chemistry of NO and related reactive species and their interactions with globins, and suggest that microbial globins have additional functions unrelated to 'NO' stresses. PMID- 10844667 TI - Carcinoembryonic antigens are targeted by diverse strains of typable and non typable Haemophilus influenzae. AB - Haemophilus influenzae (Hi), a commensal of the human respiratory mucosa, is an important cause of localized and systemic infections. We show that distinct strains belonging to typable (THi) and non-typable (NTHi) H. influenzae target human carcinoembryonic antigens (the membrane associated CEA family of cell adhesion molecules, are now termed CEACAMs). All strains of H. influenzae biogroup aegyptius (Hi-aeg) and more than 70% of THi and NTHi strains tested specifically recognize CEACAMI-Fc soluble constructs. Furthermore, transfection of Chinese hamster ovary cells with human CEACAM1 cDNA alone was sufficient for promoting Hi interactions with the transfected cells. The majority of the Hi-aeg strains tested interacted with soluble constructs containing only the N-terminal domain. In contrast, several THi and NTHi strains reacted with soluble constructs only when additional extracellular A and B domains of the receptor were present. The use of monoclonal antibodies confirmed that THi and NTHi strains also interact primarily at the N-domain. We used site-directed mutants of CEACAM1 that contained substitutions at surface exposed amino acids and a molecular model of the N-domain to identify the residues involved in interactions with Hi ligands. The studies show that a common region exposed at the CFG face of the molecule is targeted by diverse Hi strains. However, mutation at distinct sites within this area affected the interactions of distinct strains signifying the potential for tissue tropism via this receptor. Analyses of the molecular basis of interaction with human cell lines and purified CEA show that Hi strains, especially those belonging to Hi-aeg, interact with multiple CEACAMs. Because Neisseria meningitidis (Nm) strains are also known to bind at the CFG face of the receptor, we used Nm and Hi strains in co-infection experiments and demonstrate competition between these mucosal pathogens in colonization of target cells via CEACAMs. PMID- 10844668 TI - Molecular structure of bacterial endotoxin (Escherichia coli Re lipopolysaccharide): implications for formation of a novel heterogeneous lattice structure. AB - Analyses of crystals of Escherichia coli Re lipopolysaccharide (LPS) formed after storage in 1% triethylamine indicate that the LPS molecules are assembled to form a monolayered structure consisting of a novel heterogeneous lattice structure, the greater part of which is occupied by one kind of lattice (lattice I), corresponding to the acyl chain portion of lipid A, and the remainder is occupied by the other kind of lattice (lattice II), corresponding to the 3-deoxy-Dmanno octulosonic acid (dOclA) dimer and the N-acetylglucosamine disaccharide of lipid A. X-ray diffraction reveals that the type of cell is monoclinic (a = 5.53 A, b = 27.2 A, c = 6.47 A, alpha = 90 degrees, beta = 125.8 degrees, gamma = 90 degrees ). Atomic force microscopy shows that crystals consist of multiple layers; the thickness of a layer corresponds to the b-axis value, and two types of surface topographies are visualized. One, regarded as the view onto the acyl chain ends, is two-dimensional arrays of oval bodies that constitute the lattice, with the lattice constants corresponding to the a- and c-axes and the angle of beta (lattice I). The other, regarded as the view onto the dOclA dimers, is two dimensional arrays of dromedary-back-like bodies that constitute the lattice with axes of 9.0 and 10.7 A and the angle of 65 degrees formed by both axes (lattice II). Based on these results, we present the molecular model of E. coli Re LPS. PMID- 10844669 TI - PAS domain residues involved in signal transduction by the Aer redox sensor of Escherichia coli. AB - PAS domains sense oxygen, redox potential and light, and are implicated in behaviour, circadian rhythmicity, development and metabolic regulation. Although PAS domains are widespread in archaea, bacteria and eukaryota, the mechanism of signal transduction has been elucidated only for the bacterial photo sensor PYP and oxygen sensor FixL. We investigated the signalling mechanism in the PAS domain of Aer, the redox potential sensor and aerotaxis transducer in Escherichia coli. Forty-two residues in Aer were substituted using cysteine-replacement mutagenesis. Eight mutations resulted in a null phenotype for aerotaxis, the behavioural response to oxygen. Four of them also led to the loss of the non covalently bound FAD cofactor. Three mutant Aer proteins, N34C, F66C and N85C, transmitted a constant signal-on bias. One mutation, Y111C, inverted signalling by the transducer so that positive stimuli produced negative signals and vice versa. Residues critical for signalling were mapped onto a three-dimensional model of the Aer PAS domain, and an FAD-binding site and 'active site' for signal transduction are proposed. PMID- 10844670 TI - Interactions of Neisseria meningitidis with cells of the human meninges. AB - The interaction of Neisseria meningitidis with the meninges that surround and protect the brain is a pivotal event in the progression of bacterial meningitis. Two models of the human meninges were established in vitro, using (i) sections of fresh human brain and (ii) cultures of viable cells grown from human meningiomas. Neisseria meningitidis showed a specific predilection for binding to the leptomeninges and meningeal blood vessels in human brain and not to the cerebral cortex. There was a close correlation between the adherence of different Neisseria species to leptomeninges and cultured cells. The major ligand that mediated adherence was the pilus, and pilin variation modulated the interactions. The presence of Opa protein increased the association of Cap+ meningococci that expressed low-adhesive pili, but did not influence the association of high adhesive pili. In contrast, Opc did not influence the adherence of Cap+ meningococci, whereas loss of capsule was associated with a more intimate interaction between the bacteria and the meningioma cell that was not apparent with Cap+ meningococci. There was no evidence of internalization of meningococci by meningioma cells in vitro, an observation that is consistent with the barrier properties of the leptomeninges to N. meningitidis observed in vivo. PMID- 10844671 TI - A large-scale study of Yap1p-dependent genes in normal aerobic and H2O2-stress conditions: the role of Yap1p in cell proliferation control in yeast. AB - Yeast genes regulated by the transcriptional activator Yap1p were screened by two independent methods: (i) use of a LacZ-fused gene library and (ii) high-density membrane hybridization. Changes in transcriptome profile were examined in the presence and in the absence of Yap1p, as well as under normal and H2O2-mediated stress conditions. Both approaches gave coherent results, leading to the identification of many genes that appear to be directly or indirectly regulated by Yap1p. Promoter sequence analysis of target genes revealed that this regulatory effect is not always dependent upon the presence of a Yap1p binding site. The results show that the regulatory role of Yap1p is not restricted to the activation of stress response but that this factor can act as a positive or a negative regulator, both under normal and oxidative stress conditions. Among the targets, a few genes participating in growth control cascades were detected. In particular, the RPI1 gene, a repressor of the ras-cAMP pathway, was found to be downregulated by Yap1p during the early phase of growth, but upregulated in the stationary phase or after oxidative stress. PMID- 10844672 TI - The Bacillus subtilis cell division protein FtsL localizes to sites of septation and interacts with DivIC. AB - FtsL is a small bitopic membrane protein required for vegetative cell division and sporulation in Bacillus subtilis. We investigated its localization by fluorescence microscopy using a green fluorescent protein (GFP) fusion. GFP-FtsL was localized at mid-cell in vegetative cells and at the asymmetric septum in sporulating cells. We also show that FtsL forms a ring-like structure at the division site and that it remains localized at mid-cell during the whole septation process. By yeast two-hybrid analysis and non-denaturing polyacrylamide gel electrophoresis (PAGE) with purified proteins, FtsL was found to interact with another membrane-bound division protein, the FtsL-like DivIC protein. PMID- 10844673 TI - Targeted gene disruption in Candida albicans wild-type strains: the role of the MDR1 gene in fluconazole resistance of clinical Candida albicans isolates. AB - Resistance of the pathogenic yeast Candida albicans to the antifungal agent fluconazole is often caused by active drug efflux out of the cells. In clinical C. albicans strains, fluconazole resistance frequently correlates with constitutive activation of the MDR1 gene, encoding a membrane transport protein of the major facilitator superfamily that is not expressed detectably in fluconazole-susceptible isolates. However, the molecular changes causing MDR1 activation have not yet been elucidated, and direct proof for MDR1 expression being the cause of drug resistance in clinical C. albicans strains is lacking as a result of difficulties in the genetic manipulation of C. albicans wild-type strains. We have developed a new strategy for sequential gene disruption in C. albicans wild-type strains that is based on the repeated use of a dominant selection marker conferring resistance against mycophenolic acid upon transformants and its subsequent excision from the genome by FLP-mediated, site specific recombination (MPAR-flipping). This mutagenesis strategy was used to generate homozygous mdr1/mdr1 mutants from two fluconazole-resistant clinical C. albicans isolates in which drug resistance correlated with stable, constitutive MDR1 activation. In both cases, disruption of the MDR1 gene resulted in enhanced susceptibility of the mutants against fluconazole, providing the first direct genetic proof that MDR1 mediates fluconazole resistance in clinical C. albicans strains. The new gene disruption strategy allows the generation of specific knock out mutations in any C. albicans wild-type strain and therefore opens completely novel approaches for studying this most important human pathogenic fungus at the molecular level. PMID- 10844674 TI - Novel type I restriction specificities through domain shuffling of HsdS subunits in Lactococcus lactis. AB - This study identifies a natural system in Lactococcus lactis, in which a restriction modification specificity subunit resident on a 6159 bp plasmid (pAH33) alters the specificity of a functional R/M mechanism encoded by a 20.3 kb plasmid, pAH82. The new specificity was identified after phenotypic and molecular analysis of a 26.5 kb co-integrate plasmid (pAH90), which was detected after bacteriophage challenge of the parent strain. Analysis of the regions involved in the co-integration revealed that two novel hybrid hsdS genes had been formed during the co-integration event. The HsdS chimeras had interchanged the C- and N terminal variable domains of the parent subunits, generating two new restriction specificities. Comparison of the parent hsdS genes with other type I specificity determinants revealed that the region of the hsdS genes responsible for the co integration event is highly conserved among lactococcal type I hsdS determinants. Thus, as hsdS determinants are widespread in the genus Lactococcus, new restriction specificities may evolve rapidly after homologous recombination between these genes. This study demonstrates that, similar to previous observations in Gram-negative bacteria, a Gram-positive bacterium can acquire novel restriction specificities naturally through domain shuffling of resident HsdS subunits. PMID- 10844675 TI - Growth phase-dependent expression and degradation of histones in the thermophilic archaeon Thermococcus zilligii. AB - HTz is a member of the archaeal histone family. The archaeal histones have primary sequences and structural similarity to the eukaryal histone fold domain, and are thought to resemble the archetypal ancestor of the eukaryal nucleosome core histones. The effects of growth phase on the total soluble proteins from Thermococcus zilligii, isolated after various stages of growth from mid logarithmic to late stationary phase, were examined by denaturing polyacrylamide gel electrophoresis. On entry into stationary phase, at least 11 proteins were detected that changed considerably in level. One of these proteins was identified by Western hybridization as HTz. The level of HTz decreased dramatically as cells entered stationary phase, and it could not be detected by late stationary phase. Unexpectedly, the Western hybridization detected a second protein, with an estimated molecular mass of approximately 14 kDa, which paralleled the decrease in level of HTz. Native purified HTz was shown to retain complete activity after prolonged incubation at the growth temperature of the organism, suggesting that the decrease in HTz was a specific cell-regulated process. Analysis of native purified HTz by electrospray ionization mass spectrometry revealed the molecular masses of HTz1 and HTz2 to be 7204 +/- 3 Da and 7016 +/- 3 Da respectively. The only non-covalent species that was detected corresponded to the molecular mass of an HTz1-HTz2 heterodimer. Northern analyses of T. zilligii total RNA with an htz1 gene probe indicated a rapid decrease in expression of htz1 with progression of the growth phase, and complete repression of htz1 transcript synthesis by late logarithmic phase. Three proteins that changed in level with growth phase were identified by N-terminal sequence analysis. The first was homologous to a hypothetical protein conserved in all Archaea sequenced to date, the second to the Sac10b family of archaeal DNA-binding proteins and the third to the C terminal region of the leucine-responsive regulatory family of DNA-binding proteins (LRPs). PMID- 10844676 TI - Variable small protein (Vsp)-dependent and Vsp-independent pathways for glycosaminoglycan recognition by relapsing fever spirochaetes. AB - Tick-borne relapsing fever, caused by pathogenic Borrelia such as B. hermsii and B. turicatae, features recurrent episodes of bacteraemia, each of which is caused by a population of spirochaetes that expresses a different variable major protein. Relapsing fever is also associated with the infection of a variety of tissues, such as the central nervous system. In this study, we show that glycosaminoglycans (GAGs) mediate the attachment of relapsing fever spirochaetes to mammalian cells. B. hermsii strain DAH bound to immobilized heparin, and heparin and dermatan sulphate blocked bacterial binding to host cells. Bacterial binding was diminished by inhibition of host cell GAG synthesis or sulphation, or by the enzymatic removal of GAGs. GAGs mediated the attachment of relapsing fever spirochaetes to potentially relevant target cells, such as endothelial and glial cells. B. hermsii was able to attach to GAGs independently of variable major proteins, because strains expressing the variable major proteins Vsp33, Vlp7 or no variable major protein at all each recognized GAGs. Nevertheless, we found that a variable major protein of B. turicatae directly promoted GAG binding by this relapsing fever spirochaete. B. turicatae strain Oz1 serotype B, which expresses the variable major protein VspB, bound to GAGs more efficiently than did B. turicatae Oz1 serotype A, which expresses VspA. Recombinant VspB, but not VspA, bound to heparin and dermatan sulphate. Previous studies have shown that strain Oz1 serotype B grows to higher concentrations in the blood than does Oz1 serotype A. Thus, relapsing fever spirochaetes have the potential to express Vsp dependent and Vsp-independent GAG-binding activities and, for one pair of highly related B. turicatae strains, differences in GAG binding correlate with differences in tissue tropism. PMID- 10844677 TI - The thermostabilizing domain of the modular xylanase XynA of Thermotoga maritima represents a novel type of binding domain with affinity for soluble xylan and mixed-linkage beta-1,3/beta-1, 4-glucan. AB - Thermotoga maritima XynA is an extremely thermostable modular enzyme with five domains (A1-A2-B-C1-C2). Its catalytic domain (-B-) is flanked by duplicated non catalytic domains. The C-terminal repeated domains represent cellulose-binding domains (CBDs). Xylanase domains related to the N-terminal domains of XynA (A1 A2) are called thermostabilizing domains because their deletion normally leads to increased thermosensitivity of the enzymes. It was found that a glutathione-S transferase (GST) hybrid protein (GST-A1A2) containing both A-domains of XynA can interact with various soluble xylan preparations and with mixed-linkage beta 1,3/beta-1,4-glucans. GST-A1A2 showed no affinity for insoluble microcrystalline cellulose, whereas, vice versa, GST-C2, which contains the C-terminal CBD of XynA, did not interact with soluble xylan. Another hybrid protein, GST-A2, displayed the same binding properties as GST-A1A2, indicating that A2 alone can also promote xylan binding. The dissociation constants for the binding of xylose, xylobiose, xylotriose, xylotetraose and xylopentaose by GST-A2, as determined at 20 degrees C by fluorescence quench experiments, were 8.1 x 10(-3) M, 2.3 x 10( 4) M, 2.3 x 10(-5) M, 2.5 x 10(-6)M and 1.1 x 10(-6) M respectively. The A domains of XynA, which are designated as xylan binding domains (XBD), are, from the structural as well as the functional point of view, prototypes of a novel class of binding domains. More than 50 related protein segments with hitherto unknown function were detected in about 30 other multidomain beta-glycanases, among them putative plant (Arabidopsis thaliana) xylanases. It is argued that polysaccharide binding and not thermostabilization is the main function of A-like domains. PMID- 10844678 TI - Suppression of a DnaX temperature-sensitive polymerization defect by mutation in the initiation gene, dnaA, requires functional oriC. AB - Temperature sensitivity of DNA polymerization and growth, resulting from mutation of the tau and gamma subunits of Escherichia coli DNA polymerase III, are suppressed by Cs,Sx mutations of the initiator gene, dnaA. These mutations simultaneously cause defective initiation at 20 degrees C. Efficient suppression, defined as restoration of normal growth rate at 39 degrees C to essentially all the cells, depends on functional oriC. Increasing DnaA activity in a strain capable of suppression, by introducing a copy of the wild-type allele, increasing the suppressor gene dosage or introducing a seqA mutation, reversed the suppression. This suggests that the suppression mechanism depends on reduced activity of DnaACs, Sx. Models that assume that suppression results from an initiation defect or from DnaACs,Sx interaction with polymerization proteins during nascent strand synthesis are proposed. PMID- 10844679 TI - PEST sequences in cAMP-dependent protein kinase subunits of the aquatic fungus Blastocladiella emersonii are necessary for in vitro degradation by endogenous proteases. AB - During Blastocladiella emersonii germination, the regulatory (R) and the catalytic (C) subunits of the cAMP-dependent protein kinase (PKA) are rapidly and concurrently degraded, after PKA activation in response to a transient increase in intracellular cAMP levels. The possibility that PEST sequences could be acting as proteolytic recognition signals in this process was investigated, and high score PEST sequences were found in both B. emersonii R and C subunits. Deletions in the PEST sequences were obtained by site-directed mutagenesis and the different PKA subunits were independently expressed in Escherichia coli. Proteolysis assays of the various R and C recombinant forms, using B. emersonii cell extracts as the source of proteases, showed a strong correlation between the presence of high score PEST sequences and susceptibility to degradation. Furthermore, the amino-terminal sequence of the proteolytic fragments indicated that the cleavage sites in both subunits are located at or near the PEST regions. The PEST sequence in B. emersonii C subunit, which when deleted or disrupted leads to resistance to proteolysis, is entirely contained in the 72-amino-acid extension located in the N-terminus of the protein. C subunit mutants carrying deletions in this region displayed little difference in their kinetic properties or enzyme thermostability. These results suggest that the N-terminal extension may only play a role in C subunit degradation. PMID- 10844680 TI - Regulation of quorum sensing in Vibrio harveyi by LuxO and sigma-54. AB - The bioluminescent marine bacterium Vibrio harveyi controls light production (lux) by an elaborate quorum-sensing circuit. V. harveyi produces and responds to two different autoinducer signals (AI-1 and AI-2) to modulate the luciferase structural operon (luxCDABEGH) in response to changes in cell-population density. Unlike all other Gram-negative quorum-sensing organisms, V. harveyi regulates quorum sensing using a two-component phosphorylation-dephosphorylation cascade. Each autoinducer is recognized by a cognate hybrid sensor kinase (called LuxN and LuxQ). Both sensors transduce information to a shared phosphorelay protein called LuxU, which in turn conveys the signal to the response regulator protein LuxO. Phospho-LuxO is responsible for repression of luxCDABEGH expression at low cell density. In the present study, we demonstrate that LuxO functions as an activator protein via interaction with the alternative sigma factor, sigma54 (encoded by rpoN). Our results suggest that LuxO, together with sigma54, activates the expression of a negative regulator of luminescence. We also show that phenotypes other than lux are regulated by LuxO and sigma54, demonstrating that in Vibrio harveyi, quorum sensing controls multiple processes. PMID- 10844681 TI - Increased sensitivity to the antimalarials mefloquine and artemisinin is conferred by mutations in the pfmdr1 gene of Plasmodium falciparum. AB - The declining efficacy of chloroquine and pyrimethamine/sulphadoxine in the treatment of human malaria has led to the use of newer antimalarials such as mefloquine and artemisinin. Sequence polymorphisms in the pfmdr1 gene, the gene encoding the plasmodial homologue of mammalian multidrug resistance transporters, have previously been linked to resistance to chloroquine in some, but not all, studies. In this study, we have used a genetic cross between the strains HB3 and 3D7 to study inheritance of sensitivity to the structurally unrelated drugs mefloquine and artemisinin, and to several other antimalarials. We find a complete allelic association between the HB3-like pfmdr1 allele and increased sensitivity to these drugs in the progeny. Different pfmdr1 sequence polymorphisms in other unrelated lines were also associated with increased sensitivity to these drugs. Our results indicate that the pfmdr1 gene is an important determinant of susceptibility to antimalarials, which has major implications for the future development of resistance. PMID- 10844682 TI - Oligomerization of the chromatin-structuring protein H-NS. AB - H-NS is a major component of the bacterial nucleoid, involved in condensing and packaging DNA and modulating gene expression. The mechanism by which this is achieved remains unclear. Genetic data show that the biological properties of H NS are influenced by its oligomerization properties. We have applied a variety of biophysical techniques to study the structural basis of oligomerization of the H NS protein from Salmonella typhimurium. The N-terminal 89 amino acids are responsible for oligomerization. The first 64 residues form a trimer dominated by an alpha-helix, likely to be in coiled-coil conformation. Extending this polypeptide to 89 amino acids generated higher order, heterodisperse oligomers. Similarly, in the full-length protein no single, defined oligomeric state is adopted. The C-terminal 48 residues do not participate in oligomerization and form a monomeric, DNA-binding domain. These N- and C-terminal domains are joined via a flexible linker which enables them to function independently within the context of the full-length protein. This novel mode of oligomerization may account for the unusual binding properties of H-NS. PMID- 10844683 TI - Cell division, guillotining of dimer chromosomes and SOS induction in resolution mutants (dif, xerC and xerD) of Escherichia coli. AB - We have studied the growth and division of xerC, xerD and dif mutants of Escherichia coli, which are unable to resolve dimer chromosomes. These mutants express the Dif phenotype, which includes reduced viability, SOS induction and filamentation, and abnormal nucleoid morphology. Growth was studied in synchronous cultures and in microcolonies derived from single cells. SOS induction and filamentation commenced after an apparently normal cell division, which sheared unresolved dimer chromosomes. This has been called guillotining. Microcolony analysis demonstrated that cell division in the two daughter cells was inhibited after guillotining, and microcolonies formed that consisted of two filaments lying side by side. Growth of these filaments was severely reduced in hipA+ strains. We propose that guillotining at dif destroys the expression of the adjacent hipBA genes and, in the absence of continued formation of HipB, HipA inhibits growth. The length of the filaments was also affected by SfiA: sfiA dif hipA mutants initially formed filaments, but cell division at the ends of the filaments ultimately produced a number of DNA-negative cells. If SOS induction was blocked by lexA3 (Ind-), filaments did not form, and cell division was not inhibited. However, pedigree analysis of cells in microcolonies demonstrated that lethal sectoring occurred as a result of limited growth and division of dead cells produced by guillotining. PMID- 10844684 TI - Polynucleotide phosphorylase, RNase II and RNase E play different roles in the in vivo modulation of polyadenylation in Escherichia coli. AB - Poly(A) tails in Escherichia coli are hypothesized to provide unstructured single stranded substrates that facilitate the degradation of mRNAs by ribonucleases. Here, we have investigated the role that such nucleases play in modulating polyadenylation in vivo by measuring total poly(A) levels, polyadenylation of specific transcripts, growth rates and cell viabilities in strains containing various amounts of poly(A) polymerase I (PAP I), polynucleotide phosphorylase (PNPase), RNase II and RNase E. The results demonstrate that both PNPase and RNase II are directly involved in regulating total in vivo poly(A) levels. RNase II is primarily responsible for degrading poly(A) tails associated with 23S rRNA, whereas PNPase is more effective in modulating the polyadenylation of the lpp and 16S rRNA transcripts. In contrast, RNase E appears to affect poly(A) levels indirectly through the generation of new 3' termini that serve as substrates for PAP I. In addition, whereas excess PNPase suppresses polyadenylation by more than 70%, the toxicity associated with increased poly(A) levels is not reduced. Conversely, toxicity is significantly reduced in the presence of excess RNase II. Overproduction of RNase E leads to increased polyadenylation and no reduction in toxicity. PMID- 10844685 TI - Molecular basis of Salmonella-induced enteritis. AB - Salmonella pathogenesis is a complex and multifactorial phenomenon. Many genes required for full virulence in mice have been identified, but only a few of these have been shown to be necessary for the induction of enteritis. Likewise, at least some of the Salmonella virulence factors affecting enteritis do not appear to be required for infection of systemic sites in mice. This suggests that subsets of virulence genes influence distinct aspects of Salmonella pathogenesis. Recently, considerable progress has been made in characterizing the virulence mechanisms influencing enteritis caused by non-typhoid Salmonella spp. The Salmonella pathogenicity island-1-encoded type III secretion system mediates the translocation of secreted effector proteins into target epithelial cells. These effector proteins are key virulence factors required for Salmonella intestinal invasion and the induction of fluid secretion and inflammatory responses. PMID- 10844686 TI - Host adaptation and the emergence of infectious disease: the Salmonella paradigm. AB - The recent emergence of food-borne pathogens, such as Salmonella enterica serotype Enteritidis (S. enteritidis) and Escherichia coli O157:H7, has generated increasing interest in how infectious diseases can invade, persist and spread within new host populations. To alter their host range pathogens require adaptations, which ensure their circulation in a new animal population. Adaptations for circulation in different populations of vertebrate hosts seem to have been acquired multiple times within the genus Salmonella because extant Salmonella serotypes differ greatly with regard to host range. In this article, mechanisms involved in host adaptation are deduced by considering the influence of the host immune response on circulation of Salmonella serotypes within populations of vertebrate animals. This approach contributes to the identification of genes involved in host adaptation and provides new insights into the emergence of food-borne pathogens. PMID- 10844687 TI - Salmonella pathogenicity island 2. AB - Systemic infections by Salmonella enterica, such as typhoid fever, are a significant threat to human health. Recent studies indicate that the function of a type III secretion system encoded by Salmonella Pathogenicity Island 2 (SPI2) is central for the ability of S. enterica to cause systemic infections and for intracellular pathogenesis. This review summarizes approaches leading to the identification of SPI2, the molecular genetics and evolution of SPI2, and the current understanding of the regulation of gene expression. Recent studies have indicated that SPI2 is used by intracellular Salmonella to actively modify functions of the host cells. The role of SPI2 during pathogenesis of salmonellosis and current models regarding function will be discussed. PMID- 10844688 TI - Unravelling the mysteries of virulence gene regulation in Salmonella typhimurium. AB - Salmonella typhimurium, which causes gastroenteritis in calves and humans as well as a typhoid-like disease in mice, uses numerous virulence factors to infect its hosts. Genes encoding these factors are regulated by many environmental conditions and regulatory pathways in vitro. Many virulence genes are specifically induced at particular sites during infection or in cultured host cells. The complex regulation of virulence genes observed in vitro may be necessary to restrict their expression to specific locations within the host. In vitro and in vivo studies provide clues about how virulence genes might be regulated in vivo. Future studies must assess the actual environmental signals and regulators that modulate each virulence gene in vivo and determine how multiple regulatory pathways are integrated to co-ordinate the appropriate expression of virulence factors at specific sites in vivo. PMID- 10844689 TI - Subcellular localization of Dna-initiation proteins of Bacillus subtilis: evidence that chromosome replication begins at either edge of the nucleoids. AB - We examined the intracellular distribution of Bacillus subtilis Dna-initiation proteins by immunofluorescence microscopy to visualize the initiation complex of replication in vivo. DnaA was distributed throughout the cytoplasm, but both DnaB and DnaI were always detected as foci during the cell-division cycle. Interaction of DnaI with the DnaC helicase by the yeast two-hybrid assay suggests that DnaI acts as a helicase loader. The number of DnaB and DnaI foci within the cell exceeded that of oriC. Although the foci were not always co-localized with oriC, they seemed to be localized near the outer or inner edges of the nucleoids at initiation of replication. When the replication cycle was synchronized in cells using a temperature-sensitive dnaA mutant, duplication of the oriC region was observed predominantly near an edge of the nucleoid. Before initiation occurred, each one of the DnaB and DnaI foci was frequently observed near there. Furthermore, DnaX-GFP (DnaX is a component of DNA polymerase III) foci were detected near either of the edges of the nucleoids at the onset of replication. These results suggest that the replisome is recruited into oriC near either edge of the nucleoids to initiate chromosome replication in B. subtilis. PMID- 10844690 TI - Frequent interspecific genetic exchange between commensal Neisseriae and Neisseria meningitidis. AB - Natural sequence variation was investigated among serogroup A subgroup IV-1 Neisseria meningitidis isolated from diseased patients and healthy carriers in The Gambia, West Africa. The frequencies of DNA import were analysed by sequencing fragments of four linked genes encoding the immunogenic outer membrane proteins TbpB (transferrin binding protein B) and OpaA (an adhesin) plus two housekeeping enzymes. Seventeen foreign tbpB alleles were independently imported into the 98 strains tested, apparently due to immune selection. The median size of the imported DNA fragments was 5 kb, resulting in the occasional concurrent import of linked housekeeping genes by hitchhiking. Sequences of tbpB from other strains of N. meningitidis as well as commensal Neisseria lactamica and Neisseria spp. isolated from the same geographical area revealed that these species share a common tbpB gene pool and identified several examples of interspecific genetic exchange. These observations indicate that recombination can be more frequent between related species than within a species and indicate that effective vaccination against serogroup B meningococcal disease may be difficult to achieve. PMID- 10844691 TI - Gonococcal lipooligosaccharide is a ligand for the asialoglycoprotein receptor on human sperm. AB - In the present study, we show that Neisseria gonorrhoeae lipooligosaccharide (LOS) can bind to the asialoglycoprotein receptor (ASGP-R) on human sperm. This work demonstrates the presence of ASGP-R on human sperm. Binding of purified ASGP R ligand decreased in the presence of gonococci. Binding of purified iodinated gonococcal LOS identified a protein of molecular weight corresponding to that of human ASGP-R. The presence of excess unlabelled LOS blocked binding of iodinated gonococcal LOS. Binding of wild-type gonococcal LOS to sperm was higher than that of mutant LOS lacking the galactose ligand for ASGP-R. These data suggest that the ASGP-R on human sperm cells recognizes and binds wild-type gonococcal LOS. This interaction may contribute to the transmission of gonorrhea from infected males to their sexual partners. PMID- 10844692 TI - Identification of the urease operon in Helicobacter pylori and its control by mRNA decay in response to pH. AB - We investigated the transcription of the urease gene cluster ureABIEFGH in Helicobacter pylori to determine the regulation of gene expression of the highly produced enzyme urease. Northern blot hybridization analysis demonstrated that cells of the wild-type strain grown in an ordinary broth had transcripts of ureAB, ureABI, ureI, ureIE' and ure'FGH, but cells of a ureI-disrupted mutant had only the ureAB transcript. When the wild-type cells were exposed to pH 8 for 30 min, very little mRNA was detected. However, when exposed to pH 6, a large amount of the ureIE" transcript, which was longer than the ureIE' transcript, together with the additional transcripts ureABIEFGH and ure'EFGH were detected. Rifampicin addition experiments demonstrated that urease mRNAs, and the ureIE' transcripts in particular, are more stable at pH 5.5 than at pH 7. In accord with these results, urease activity in the crude cell extract of the pH 5.5 culture was twice as much as that of the pH 7 culture, although the amounts of UreA and UreB detected by immunoblot analysis were similar. The transcription start point of ureI was identified by primer extension using a ureA promoter-deleted mutant, and a consensus sequence of RpoD-RNA polymerase was found in the ureI promoter. The 3' end of the ureIE" mRNA, determined using S1 nuclease mapping, revealed that the transcript is able to cover the majority of the ureE open reading frame (ORF) that might be sufficient for UreE activity. Based on the above results, we conclude that the urease gene cluster of H. pylori consists of two operons, ureAB and ureIEFGH, and that primary transcripts of the latter as well as the read through transcript, ureABIEFGH, are cleaved to produce several species of mRNA. It has been suggested that the ureIEFGH operon is regulated post transcriptionally by mRNA decay in response to environmental pH. We are tempted to speculate that the ureE" transcript present in acidic pH may contribute to produce an active product that can proceed the nickel incorporation to the active centre, the final step of urease biosynthesis. PMID- 10844693 TI - The NRAMP proteins of Salmonella typhimurium and Escherichia coli are selective manganese transporters involved in the response to reactive oxygen. AB - NRAMPs (natural resistance-associated macrophage proteins) have been characterized in mammals as divalent transition metal transporters involved in iron metabolism and host resistance to certain pathogens. The mechanism of pathogen resistance is proposed to involve sequestration of Fe2+ and Mn2+, cofactors of both prokaryotic and eukaryotic catalases and superoxide dismutases, not only to protect the macrophage against its own generation of reactive oxygen species, but to deny the cations to the pathogen for synthesis of its protective enzymes. NRAMP homologues are also present in bacteria. We report the cloning and characterization of the single NRAMP genes in Escherichia coli and Salmonella enterica ssp. typhimurium, and the cloning of two distinct NRAMP genes from Pseudomonas aeruginosa and an internal fragment of an NRAMP gene in Burkholderia cepacia. The genes are designated mntH because the two enterobacterial NRAMPs encode H+-stimulated, highly selective manganese(II) transport systems, accounting for all Mn2+ uptake in each species under the conditions tested. For S. typhimurium MntH, the Km for 54Mn2+ ( approximately 0.1 microM) was pH independent, but maximal uptake increased as pH decreased. Monovalent cations, osmotic strength, Mg2+ and Ca2+ did not inhibit 54Mn2+ uptake. Ni2+, Cu2+ and Zn2+ inhibited uptake with Kis greater than 100 microM, Co2+ with a Ki of 20 microM and Fe2+ with a Ki that decreased from 100 microM at pH 7. 6 to 10 microM at pH 5.5. Fe3+ and Pb2+ inhibited weakly, exhibiting Kis of 50 microM, while Cd2+ was a potent inhibitor with a Ki of about 1 microM. E. coli MntH had a similar inhibition profile, except that Kis were three- to 10-fold higher. Both S. typhimurium and E. coli MntH also transport 55Fe2+ however, the Kms are equivalent to the Kis for Fe2+ inhibition of Mn2+ uptake, and are thus too high to be physiologically relevant. In both S. typhimurium and E. coli, mntH:lacZ constructs were strongly induced by hydrogen peroxide, weakly induced by EDTA and unresponsive to paraquat, consistent with the presence of Fur and OxyR binding sites in the promoters. Strains overexpressing mntH were more susceptible to growth inhibition by Mn2+ and Cd2+ than wild type, and strains lacking a functional mntH gene were more susceptible to killing by hydrogen peroxide. In S. typhimurium strain SL1344, mntH mutants showed no defect in invasion of or survival in cultured HeLa or RAW264.7 macrophage cells; however, expression of mntH:lacZ was induced severalfold by 3 h after invasion of the macrophages. S. typhimurium mntH mutants showed only a slight attenuation of virulence in BALB/c mice. Thus, the NRAMP Mn2+ transporter MntH and Mn2+ play a role in bacterial response to reactive oxygen species and possibly have a role in pathogenesis. PMID- 10844694 TI - Identification of a major facilitator protein from Escherichia coli involved in efflux of metabolites of the cysteine pathway. AB - A chromosomal fragment has been identified in a gene bank from Escherichia coli, which augmented the yield of cysteine in an industrial production strain. Subcloning and genetic analysis showed that an open reading frame coding for a product of 299 amino acids (Orf299) was responsible. Orf299 was synthesized in the T7 polymerase/promoter system and exhibited the properties of an integral membrane protein. Mutational interruption of orf299 did not cause a distinct phenotype; however, transformants overexpressing orf299 had lost the ability to grow in minimal medium unless it was supplemented with a source of reduced sulphur compounds, and they excreted considerable amounts of cysteine and O acetyl-L-serine, especially in the presence of thiosulphate. Most of the cysteine was found to be masked in 2-methyl-2,4-thiazolidinedicarboxylic acid. N-acetyl-L serine was also present in the medium, but it is open to question whether it represents a primary excretion product. Measurement of the induction status of the cysteine regulon by means of a cysK'-'lacZ gene fusion demonstrated that the regulon is not induced upon growth in the presence of a poor sulphur source and that the introduction of a constitutive cysB allele alleviates this deficiency. The results indicate that orf299 codes for an export pump for different metabolites of the cysteine pathway. Its relation to other efflux systems and the physiological role are discussed. PMID- 10844695 TI - Sialic acid metabolism's dual function in Haemophilus influenzae. AB - Many bacterial commensals and pathogens use the sialic acids as carbon and nitrogen sources. In Escherichia coli, the breakdown of these sugars is catalysed by gene products of the nan (Nacylneuraminate) operon; other microorganisms may use a similar catabolic strategy. Despite the known ligand and antirecognition functions of the sialic acids, the contribution of their catabolism to infection or host colonization has never been directly investigated. We addressed these questions with Haemophilus influenzae type b, which metabolizes relatively few carbohydrates, using the infant-rat infection model. The predicted H. influenzae homologue (HI0142) of the E. coli sialic acid aldolase structural gene, nanA, was subcloned and mutagenized by insertion of a kanamycin resistance cassette. Phenotypic investigation of the resulting H. influenzae aldolase mutants showed that: (i) HI0142 is essential for sialic acid degradation; (ii) the products of the open reading frames (ORFs) flanking HI0142 (HI0140, 41, 44 and 45) are likely to have the same functions as those of their counterparts in E. coli; (iii) sialylation of the lipooligosaccharide (LOS) epitope recognized by monoclonal antibody 3F11 is dependent on an environmental source of sialic acid; (iv) a nanA mutant hypersialylates its LOS sialyl acceptor, corresponding to an apparent increased fitness of the mutant in the infant-rat model; and (v) expression of the LOS sialyl acceptor is altered in cells grown without exogenous sialic acid, indicating the direct or indirect effect of sialic acid metabolism on LOS antigenicity. Taken together the data show the dual role of sialic acid catabolism in nutrition and cell surface modulation. PMID- 10844696 TI - Antisense RNA-dependent transcription termination sites that modulate lysogenic development of satellite phage P4. AB - In the lysogenic state, bacteriophage P4 prevents the expression of its own replication genes, which are encoded in the left operon, through premature transcription termination. The phage factor responsible for efficient termination is a small, untranslated RNA (CI RNA), which acts as an antisense RNA and controls transcription termination by pairing with two complementary sequences (seqA and seqC) located within the leader region of the left operon. A Rho dependent termination site, timm, was previously shown to be involved in the control of P4 replication gene expression. In the present study, by making use of phage PhiR73 as a cloning vector and of suppressor tRNAGly as a reporter gene, we characterized two additional terminators, t1 and t4. Although transcription termination at neither site requires the Rho factor, only t1 has the typical structure of a Rho-independent terminator. t1 is located between the PLE promoter and the cI gene, whereas t4 is located between cI and timm. Efficient termination at t1 requires the CI RNA and the seqA target sequence; in vitro, the CI RNA enhanced termination at t1 in the absence of any bacterial factor. A P4 mutant, in which the t1 terminator has been deleted, can still lysogenize both Rho+ and Rho- strains and exhibits increased expression of CI RNA. These data indicate that t1 and the Rho-dependent timm terminators are not essential for lysogeny. t1 is involved in CI RNA autoregulation, whereas t4 appears to be the main terminator necessary to prevent expression of the lytic genes in the lysogenic state. PMID- 10844697 TI - S-adenosylmethionine decarboxylase of Bacillus subtilis is closely related to archaebacterial counterparts. AB - Bacillus subtilis synthesizes polyamines by decarboxylating arginine to agmatine, which is subsequently hydrolysed to putrescine. Spermidine is synthesized from putrescine and decarboxylated S-adenosylmethionine (dAdoMet). In Gram-negative bacteria and in eukaryotes, AdoMet is decarboxylated by an unusual 'pyruvoyl' AdoMet decarboxylase (SpeD), the catalytic pyruvoyl moiety of which is generated by serinolysis of an internal serine with self-cleavage of the protein at the upstream peptide bond. Neither the Gram-positive bacterial nor the archaeal counterpart of the Escherichia coli SpeD enzyme were known. We have identified the corresponding B. subtilis speD gene (formely ytcF). Heterologous expression of the cognate Methanococcus jannaschii protein, MJ0315, demonstrated that it displays the same activity as B. subtilis SpeD, indicating that spermidine biosynthesis in Gram-positive bacteria and in archaea follows a pathway very similar to that of Gram-negatives and eukarya. In B. subtilis, transcription of speD is modulated by spermidine and methionine. Its expression is high under usual growth conditions. In contrast, the SpeD protein self-cleaves slowly in vitro, a noticeable difference with its archaeal counterpart. Under certain growth conditions (minimal medium containing succinate and glutamate as a carbon source), speD is co-transcribed with gapB, the gene encoding glyceraldehyde-3 phosphate dehydrogenase, an enzyme required for gluconeogenesis. This observation may couple polyamine metabolism to sulphur and carbon metabolism by a so far unknown mechanism. PMID- 10844698 TI - Interacting interfaces of the P4 antirepressor E and the P2 immunity repressor C. AB - Antirepressors have been identified as proteins interacting with transcriptional repressors leading to expression of the repressed genes. The defective satellite phage/plasmid P4 has the capacity to derepress the unrelated prophage P2 after infection, thereby getting access to the late functions of the helper that are required for P4 lytic growth. The derepression of prophage P2 is mediated by the P4 E protein that function as an antirepressor by binding to the P2 immunity repressor C. A P2 mutant, sos, has been isolated that is insensitive to the action of the P4 E protein. In the present study, we show that sos is a point mutation in the P2 immunity repressor gene C and that it makes P4 E unable to turn the transcriptional switch of P2 from the lysogenic state to the lytic mode in a two plasmid reporter system. Furthermore, the interaction between C and E, when analysed in the yeast two-hybrid system, is blocked by the sos mutation. An analysis of C mutants indicates that the dimerization function of C is located in the C-terminal part of the protein and the dimerization defective mutants are unable to bind to their operator DNA. The sos mutation does not affect the capacity of the protein to dimerize. Using the yeast two-hybrid system, compensatory E mutants have been isolated that can interact with Sos, but they are unable to turn the transcriptional switch controlled by the Sos repressor. However, one point mutation in the E protein is shown to be unable to turn the transcriptional switch controlled by the wild-type C repressor. PMID- 10844699 TI - Glycosylation deficiency phenotypes resulting from depletion of GDP-mannose pyrophosphorylase in two yeast species. AB - The genes encoding GDP-mannose pyrophosphorylase from Saccharomyces cerevisiae (SRB1/PSA1) and Candida albicans (CaSRB1) were expressed under the control of the tightly regulated promoters of MET3 and CaMET3 respectively. Northern analysis showed that the addition of methionine effectively blocks the transcription of pMET3-SRB1/PSA1 and pCaMET3CaSRB1 expression cassettes, which had been integrated into the genomes of appropriate mutants. Methionine-mediated repression of CaSRB1 caused loss of viability in C. albicans, demonstrating that, as in S. cerevisiae, the gene is essential for growth. Depletion of GDP-mannose pyrophosphorylase had a highly pleiotropic effect in the two yeasts. The major phenotypes observed were lysis, failure of cell separation and/or cytokinesis, impaired bud growth and bud's site selection, clumping and flocculation, as well as increased sensitivity to a wide range of antifungal drugs and cell wall inhibitors, and impaired hyphal switching ability. These phenotypes resulted from defects in glycosylation, as demonstrated by reduced affinity for Alcian blue and sensitivity to hygromycin B. Our results provide new information about the roles of protein glycosylation in yeast and, in particular, the steps that require GDP-mannose in the fungal pathogen C. albicans. PMID- 10844700 TI - A single glutaredoxin or thioredoxin gene is essential for viability in the yeast Saccharomyces cerevisiae. AB - Glutaredoxins and thioredoxins are small heat-stable oxidoreductases that have been conserved throughout evolution. The yeast Saccharomyces cerevisiae contains two gene pairs encoding cytoplasmic glutaredoxins (GRX1, GRX2) and thioredoxins (TRX1, TRX2). We report here that the quadruple trx1 trx2 grx1 grx2 mutant is inviable and that either a single glutaredoxin or a single thioredoxin (i.e. grx1 grx2 trx1, grx1 grx2 trx2, grx1 trx1 trx2, grx2 trx1 trx2) is essential for viability. Loss of both thioredoxins has been reported previously to lead to methionine auxotrophy consistent with thioredoxins being the sole reductants for 3'-phosphoadenosine 5'-phosphosulphate reductase (PAPS) in yeast. However, we present evidence for the existence of a novel yeast hydrogen donor for PAPS reductase, as strains lacking both thioredoxins assimilated sulphate under conditions that minimized the generation of reactive oxygen species (low aeration and absence of functional mitochondria). In addition, the assimilation of [35S] sulphate was approximately 60-fold higher in the trx1 trx2 grx1 and trx1 trx2 grx2 mutants compared with the trx1 trx2 mutant. Furthermore, in contrast to the trx1 trx2 mutant, the trx1 trx2 grx2 mutant grew on minimal agar plates, and the trx1 trx2 grx1 mutant grew on minimal agar plates under anaerobic conditions. We propose a model in which the novel reductase activity normally functions in the repair of oxidant-mediated protein damage but, under conditions that minimize the generation of reactive oxygen species, it can serve as a hydrogen donor for PAPS reductase. PMID- 10844701 TI - Saturation mutagenesis of the haloarchaeal bop gene promoter: identification of DNA supercoiling sensitivity sites and absence of TFB recognition element and UAS enhancer activity. AB - Transcription from the bop promoter in the haloarchaeon Halobacterium NRC-1, is highly induced under oxygen-limiting conditions. A DNA gyrase inhibitor, novobiocin, was previously shown to block bop gene induction and suggested that DNA supercoiling mediates transcriptional induction. A region of non-B structure was found 3' to the TATA box within an 11 bp alternating purine-pyrimidine sequence (RY box), which correlated to both increased DNA supercoiling and transcriptional induction. Here, saturation mutagenesis of the RY box region has been used to show that single-base substitutions of A(r)G either 23 or 19 bp 5' to the transcription start site temper the effect of DNA supercoiling based on novobiocin insensitivity of transcription. Mutagenesis of the region 5' to the TATA box showed its involvement in DNA supercoiling modulation of transcription, defined the 3' end of the upstream activator sequence (UAS) regulatory element, and ruled out the requirement for a TFB (TFIIB) Recognition Element. Spacing between the TATA box and UAS was found to be critical for promoter activity because insertion of partial or whole helical turns between the two elements completely inhibited transcription indicating that the UAS element does not function as a transcriptional enhancer. The results are discussed in the context of DNA melting and flexibility around the TATA box region and the involvement of multiple regulatory and transcription factors in bop promoter activity. PMID- 10844702 TI - Is gene expression in Halobacterium NRC-1 regulated by multiple TBP and TFB transcription factors? PMID- 10844703 TI - Statistical Methodology in Alzheimer's Disease Research. Proceedings of a conference. Lexington, Kentucky, USA. 15-17 May 1998. PMID- 10844704 TI - Measurement of progression in Alzheimer's disease: a clinician's perspective. AB - Patients with Alzheimer's disease (AD) and their families must confront two fundamental truths. First, AD is a uniformly progressive disease that ultimately results in debilitating cognitive impairment. Second, although there is now evidence that some medications may produce transient improvement or possibly even slowing of disease progression, there is currently no way to halt the progression of AD. Consequently, patients and their families consistently ask the following questions: 1. What new management issues can be anticipated, and when? 2. What clinical developments are atypical and merit evaluation for a superimposed problem? 3. Is the current treatment working? These questions can only be answered by referring to the natural course of AD, and specifically, information regarding measures of functional impairment and how they change over time. The information that is currently available on this topic is limited and often embodies implicit assumptions that have not been adequately tested. This information will be reviewed, and directions for future research will be outlined. PMID- 10844705 TI - Longitudinal studies of cognitive, functional and behavioural change in patients with Alzheimer's disease. AB - This paper reviews data on the natural history of symptoms in patients with Alzheimer's disease (AD) and describes some of the problems encountered in analysing longitudinal data in this population. Data on cognition, functional ability and psychiatric or behavioural symptoms have all been obtained from AD patients. Because of attrition, the length of follow-up is not uniform for all patients and neither is the frequency of evaluation. Furthermore, patients enter longitudinal studies with a wide range of symptom severity and longitudinal decline in cognition and function is distinctly non-linear. Behavioural symptoms do not progress regularly in AD but are episodic phenomena not closely related to cognition or function. Strengths and limitations of various analytic techniques used for hypothesis testing with these longitudinal data are described. PMID- 10844706 TI - Reliability of cognitive tests used in Alzheimer's disease. AB - Assessment of cognitive status is a key component of monitoring Alzheimer's patients during the course of their illness. The reliability of a cognitive test is a measure of its reproducibility under replicate conditions. In the classical setting, reliability is defined in three ways: the ratio of the variance of the true scores to the variance of the observed scores; the correlation of observed scores on two parallel forms of the test, and the square of the correlation between the observed score and the true score. In the classical case of independence of true scores and measurement errors, the three definitions are equivalent. Estimation of reliability through analysis of variance techniques and construction of confidence intervals is accomplished when the true scores and errors are normally distributed. This paper examines a non-parametric, probabilistic estimate of reliability as the probability that, given a parallel test, the second set of scores has the same ranking as the first set. In the classical case there is a monotonic relationship between this measure and the reliability. This measure is also linked to Kendall's tau. The performance of the probabilistic measure is compared with the traditional measures in a variety of models, including those with bounded scales, and those with skewed distributions. The ideas are extended to the case of the reliability of change scores and to biased estimators of true scores. In this context truncation models and Bayes estimates of true scores are considered. PMID- 10844707 TI - Measuring cognitive change in a cohort of patients with Alzheimer's disease. AB - Annualized rates of cognitive change in Alzheimer's disease (AD), an important index of disease progression, show marked variability. To determine factors leading to such variability, we computed rates of change in a cohort of patients with AD tested annually with the Mini Mental State Examination (MMSE) and the more detailed Dementia Rating Scale (DRS). Estimates of rates of change (slopes) and intercepts were calculated using least squares and best linear unbiased predictors (BLUPs). Potential predictors of rates of change were examined using multivariate linear regression analysis. We found that the MMSE had more noise than the DRS. For the MMSE, slopes showed a moderate floor effect and a slight ceiling, depending on initial MMSE scores. These effects were less prominent for the DRS, for which slopes increased as intercepts decreased. In analyses of predictors of change, the MMSE was less useful than the DRS. In multiple linear regression models using DRS data, predictors showed statistically stronger effects and explained a greater extent of variation of slopes than did similar models using MMSE data. For example, among patients who died and underwent brain examination at autopsy, neuropathology of Lewy bodies plus AD (Lewy Body variant; LBV) was associated with significantly faster rates of cognitive decline compared to pure AD in analyses that used the DRS, but only trends were identified with the MMSE. The metric properties and longitudinal characteristics of cognitive tests and the statistical methods used to calculate change are key factors in obtaining reliable estimates of change in studying cohorts of patients with AD. PMID- 10844708 TI - Analysis of longitudinal data in an Alzheimer's disease clinical trial. AB - Evidence of delayed progression is the primary mechanism for demonstrating therapeutic efficacy in clinical trials in Alzheimer's disease. In the major trials of therapeutic treatment of AD, to date, measures based on clinical judgement and cognitive performance, instead of mortality, have been used as the primary response measures. There is good reason for this since the course of the disease is quite long, and AD trials designed around mortality would require either very large sample sizes or very long follow-up in order to have adequate power. However, the evaluation of progression in AD using clinical markers is subject to a number of challenges often found in longitudinal databases, for example, missing data, floor and ceiling effects and non-linearity. Unfortunately, few of these issues are being addressed in the typical analysis of progression data. This paper explores these analytic issues in the context of the recently completed Alzheimer's Disease Cooperative Study trial of vitamin E and Selegeline in moderate AD patients. PMID- 10844709 TI - Blomqvist revisited: how and when to test the relationship between level and longitudinal rate of change. AB - Longitudinal studies are often interested in assessing the relationship between severity (level) and rate of change (slope). Blomqvist describes an estimator of this relationship that has been used in a variety of contexts. This paper reviews and generalizes the Blomqvist method. Most published applications of the Blomqvist method contain substantial bias because they fail to consider and accommodate confounding due to the pooling of multiple age cohorts in a single analysis. We describe this bias, and present an unbiased algorithm consistent with the intentions of Blomqvist. We also explore when it is appropriate to apply the Blomqvist analysis, and what inferences can be made using this statistic. Aetiological inference about premorbid level of function predicting future rate of decline is often desired, but may not be justified when modelling chronic progressive conditions, since differential progression prior to the start of longitudinal follow-up can induce a relationship between level and rate of decline, even in the absence of an aetiologically relevant association. We conclude that aetiological inference by the Blomqvist analysis is not appropriate in most investigations of chronic progressive disease. Using the model to develop descriptive and predictive equations in these circumstances, however, remains appropriate, as does testing simply for clinical heterogeneity in longitudinal rate of decline. PMID- 10844710 TI - Promoting interactions with basic scientists and clinicians: the NIA Alzheimer's Disease Data Coordinating Center. AB - To benefit Alzheimer's disease research, a central data co-ordinating centre (CDCC) is planned that will systematically collect data from 27 Alzheimer's disease centres (ADCs) located nationwide. This CDCC will combine, analyse and disseminate epidemiologic, demographic, clinical and neuropathological data to researchers from the ADCs and the broader scientific community. New and larger scale collaborative studies on Alzheimer's disease will be possible through this centre. Since 1 July 1997, an interim data co-ordinating centre (IDCC) has been serving as the agent of the ADCs to begin the data sharing process until a permanent CDCC is established. The data collected by the IDCC are limited to administrative information and to indexing of specimens and clinical material, with future plans for the transfer of the data collected to the CDCC once it is established. PMID- 10844711 TI - Promoting interactions with basic scientists and clinicians: summary of the panel session. AB - This is a summary of the main topics discussed in a panel session on the theme of promoting interactions with basic scientists and clinicians. The main topics discussed were: communicating with basic scientists and clinicians; models of collaboration; manuscript review at an Alzheimer's disease centre, and suggestions for conducting future forums or conferences on methodology encountered in Alzheimer's disease. PMID- 10844712 TI - Community-based studies of Alzheimer's disease: statistical challenges in design and analysis. AB - Alzheimer's disease is a chronic disease, primarily of the elderly, characterized by progressive dementia and eventual death. Community-based studies will likely provide a better representation of the spectrum of disease than will studies drawn solely from clinical sources, because an unknown and possibly substantial fraction of the cases do not come to the attention of the medical care system, or are diagnosed only very late in the disease. Community-based studies will provide not only more accurate estimates of prevalence and incidence, but also more directly comparable unaffected people for studies of risk factors for onset and progression. Such studies are likely to consist of a census component where relatively inexpensive but useful auxiliary information is collected and a probability sample from the census, with the detailed and costly clinical diagnosis of Alzheimer's disease restricted to the sample. The statistician faces challenges both in designing a sample that meets multiple objectives efficiently and in analysing data from the resulting complex survey designs. PMID- 10844713 TI - Methods for projecting the incidence and prevalence of chronic diseases in aging populations: application to Alzheimer's disease. AB - Projections of the incidence and prevalence of disease are important for public health planning. This paper describes methods for projecting the incidence and prevalence of a chronic disease in ageing populations. The approach uses age specific disease incidence rates together with assumptions about survival to reconstruct disease prevalence. The methods can be used to evaluate the potential impact of public health interventions that may prevent disease or prolong survival. We used the methods to project the future prevalence of Alzheimer's disease in the United States. We found that the prevalence of Alzheimer's disease will nearly quadruple over the next 50 years. Although projections of the absolute prevalence are sensitive to assumptions about the age-specific incidence rates of disease, the proportionate growth is relatively insensitive. The increase in prevalence results from the ageing of the U.S. population. In order to perform the calculations, we have assembled U.S. Census population projections and U.S. mortality rates into computer software that is available from the authors at www.jhsph.edu/Departments/Biostats/software.h tml. PMID- 10844714 TI - Computing estimates of incidence, including lifetime risk: Alzheimer's disease in the Framingham Study. The Practical Incidence Estimators (PIE) macro. AB - The incidence of disease is estimated in medical and public health applications using various different techniques presented in the statistical and epidemiologic literature. Many of these methods have not yet made their way to popular statistical software packages and their application requires custom programming. We present a macro written in the SAS macro language that produces several estimates of disease incidence for use in the analysis of prospective cohort data. The development of the Practical Incidence Estimators (PIE) Macro was motivated by research in Alzheimer's Disease (AD) in the Framingham Study in which the development of AD has been prospectively assessed over an observation period of 24 years. The PIE Macro produces crude and age-specific incidence rates, overall and stratified by the levels of a grouping variable. In addition, it produces age-adjusted rates using direct standardization to the combined group. The user specifies the width of the age groups and the number of levels of the grouping variable. The PIE macro produces estimates of future risk for user defined time periods and the remaining lifetime risk conditional on survival event-free to user-specified ages. This allows the user to investigate the impact of increasing age on the estimate of remaining lifetime risk of disease. In each case, the macro provides estimates based on traditional unadjusted cumulative incidence, and on cumulative incidence adjusted for the competing risk of death. These estimates and their respective standard errors, are provided in table form and in an output data set for graphing. The macro is designed for use with survival age as the time variable, and with age at entry into the study as the left-truncation variable; however, calendar time can be substituted for the survival time variable and the left-truncation variable can simply be set to zero. We illustrate the use of the PIE macro using Alzheimer's Disease incidence data collected in the Framingham Study. PMID- 10844715 TI - Estimating odds ratios adjusting for misclassification in Alzheimer's disease risk factor assessment. AB - Epidemiological studies of Alzheimer's disease and dementia are often two-phase studies including a screening phase and a clinical assessment phase. It is common to interview a relative of the subject at each of these phases to obtain information about the subject's exposure to risk factors. This can result in a misclassification error when assessing risk factors, as the two responses of the relative often differ. This is especially a problem for risk factors involving life-style and family history which cannot be confirmed using the subject's medical records. A naive analysis using data from each phase separately would give two different estimates of the odds ratio; both estimates could be biased. In this paper, we extend the estimation methods adjusting for misclassification developed by Liu and Liang to data collected through two-phase sampling. We first use a latent class analysis and the EM algorithm to estimate the misclassification parameters. We then derive the maximum pseudo-likelihood estimators, conditional on the misclassification parameters, to estimate the odds ratios accounting for the complex sampling study design. We propose to use the jack-knife estimator for estimation of the variances. We apply the above method to data collected in the Indianapolis-Ibadan Dementia Study to estimate the odds ratio for smoking adjusting for misclassification error. PMID- 10844716 TI - Balanced centralized and distributed database design in a clinical research environment. AB - Clinical research databases can meet both research and clinical needs, but this ideal is seldom achieved. Priorities often differ for those who collect and ultimately use the data and those who develop data systems. Traditional database designs also create logistical barriers that hamper communication. The Michigan Alzheimer's Disease Research Center has developed a secure, distributed data system with centralized data entry that provides an intuitive, individually customized interface for investigators in their clinics, laboratories and offices. Data are kept in a form that can be readily understood without reference to a code-book. Investigators can modify and query their own copies of the database without knowledge of programming languages. Balancing centralized and distributed designs for research databases enhance the accuracy and completeness of data collection and increases the use of data for research and clinical care. PMID- 10844717 TI - Estimating the incidence of dementia from two-phase sampling with non-ignorable missing data. AB - Two-phase sampling designs have been used in the field of psychiatry to estimate prevalence and incidence of a rare disease such as dementia and Alzheimer's disease. In a longitudinal study on dementia, since the repeated two-phase sampling is conducted several years after the baseline wave, some subjects may die before the follow-up wave, thus their disease status prior to death is missing. There are reasons to suggest that the missing due to death is non ignorable. Estimation of disease incidence from longitudinal dementia study has to appropriately adjust for data missing by death as well as the sampling design used at each study wave. In this paper we adopt a selection model approach to model the missing data by death and use a likelihood approach to derive incidence estimates. A modified EM algorithm is used to deal with data from sampling selection. The non-parametric jack-knife variance estimator is used to derive variance estimates for the model parameters and the incidence estimates. The proposed approaches are applied to data from the Indianapolis-Ibadan Dementia Study. PMID- 10844718 TI - A change point model for estimating the onset of cognitive decline in preclinical Alzheimer's disease. AB - Dementia is characterized by accelerated cognitive decline before and after diagnosis as compared to normal ageing. Determining the time at which that rate of decline begins to accelerate in persons who will develop dementia is important both in describing the natural history of the disease process and in identifying the optimal time window for which treatments might be useful. We model that time at which the rate of decline begins to accelerate in persons who develop dementia relative to those who do not by using a change point in a mixed linear model. A profile likelihood method is proposed to draw inferences about the change point. The method is applied to data from the Bronx Ageing Study, a cohort study of 488 initially non-demented community-dwelling elderly individuals who have been examined at approximately 12-month intervals over 15 years. Cognitive function was measured using the Buschke Selective Reminding test, a memory test with high reliability and known discriminative validity for detecting dementia. We found that the rate of cognitive decline as measured by this test in this cohort increases on average 5.1 years before the diagnosis of dementia. PMID- 10844719 TI - Spacing of follow-up waves in incidence studies. AB - Longitudinal studies of dementia are often undertaken to estimate the incidence of dementia and to identify incident cases for the study of risk factors measured on the entire cohort at baseline. The power of these analyses is determined primarily by the number of demented cases identified. Increasing the number of waves of evaluation of the cohort in a given time period increases the yield of demented cases but it also increases the cost of the study. This paper provides a method for estimating dementia incidence in the presence of mortality and loss to follow-up. It also assesses the trade-off between the cost (as measured by the number of re-evaluations) and the yield of incident dementia cases so that longitudinal studies of dementia can be designed optimally in the future. PMID- 10844720 TI - Estimating incidence of dementia subtypes: assessing the impact of missed cases. AB - In many community-based studies on the incidence of dementia, a target population is screened and a subsample is clinically evaluated at baseline and follow-up. Incidence rates are affected by missed cases at both exams and this complicates the estimation of these rates. Recent work proposes a regression-based technique for joint estimation of prevalence and incidence and suggests the use of surrogate information obtained on the entire cohort at both times to calculate the expected score equation contribution for individuals missing clinical exams at one or both times. This helps to quantify the impact of missed diagnosis upon the incidence estimates and their confidence intervals. We extend this work to the setting of subtypes of dementia for use in the Honolulu-Asia Aging Study on incidence of dementia. The technique is applied using two separate models for the effect of age on dementia incidence. Subsequently, shrinkage estimation methods are applied to provide more precise estimates of the rates. Published in 2000 by John Wiley & Sons, Ltd. PMID- 10844721 TI - Modelling the decline pattern in functional measures from a prevalent cohort study. AB - In studying decline among cognitively impaired people, a prevalent cohort study design is commonly used to account for entry into the study at different levels of impairment. The data set typically consists of many short series of repeated measurements collected over time. However, the time origin, such as time of disease/impairment onset, is often uncertain. In order to model non-linear decline patterns in functional test scores and associated risk factors with such data, we propose two approaches as alternatives to Liu et al. One approach models change over adjacent visits with varying time intervals. The second models the change since baseline using a random effect for heterogeneity of change. We used these two approaches to examine the decline in cognitive test scores among special care unit (SCU) and non-SCU residents at the New York sites of the National Institute on Aging (NIA) collaborative studies of special dementia care. The analyses suggest that, controlling for several covariates, SCU residents experienced more rapid cognitive decline than did non-SCU residents. The relative advantages and disadvantages of the two models are discussed. PMID- 10844722 TI - Modelling mini mental state examination changes in Alzheimer's disease. AB - The Mini Mental State Examination (MMSE) is widely used to measure dementia severity in Alzheimer's disease patients. While changes over time in the MMSE due to dementia have been studied, the relationship between MMSE scores and the duration of disease course is less well understood. Using the Consortium to Establish a Registry for Alzheimer's Disease (CERAD) data, we modelled change in MMSE as a function of time for this population. For this purpose we used the interval between consecutive MMSE assessments as the time factor. We also investigated the impact of sex, education and age at testing on the resulting model. Analyses showed that Alzheimer's disease progression over time (ADP) can be modelled using a cubic or a logarithmic function of MMSE score. From these curves ADP can be obtained as a function of MMSE. These models demonstrate that there are different rates of change for various ranges of the MMSE. Additional analyses suggest that patient factors affect rates of ADP, younger patients and more educated patients progress more rapidly, while sex has little impact on ADP. Such estimations of disease course are useful when comparing different populations for both clinical and research purposes. PMID- 10844723 TI - Mixed effect models of longitudinal Alzheimer's disease data: a cautionary note. AB - Longitudinal studies of cognitive function in Alzheimer's disease (AD) patients are powerful tools to better understand the biology and natural history of the disease, but the attributes of the studies that make them valuable also pose special challenges to analysts. A fundamental problem is the accurate measure of time at which cognitive decline begins. Investigators typically use the date of AD diagnosis or the date of enrollment in an AD study. If the rate of cognitive decline is non-linear, variables associated with the time of diagnosis or enrollment might artificially be associated with the rate of decline. Unlike the mixed effects models typically used to analyse cognitive decline, summary measure analyses do not directly compare the rate of decline with time since decline began, and, therefore, are less sensitive to biased measures of time of decline. We simulated trajectories of cognitive decline using the multivariate normal random effect model and tested the ability of the two analytic techniques to discriminate between true and spurious associations. Our analyses suggest summary measure models are less likely to detect spurious associations generated by biased measures of time at which decline begins, and more likely to detect true associations concealed by biased time measurement. PMID- 10844724 TI - Application of item response theory for development of a global functioning measure of dementia with linear measurement properties. AB - An ideal measure of global functioning for patients with dementia would discriminate at very high and very low levels of functioning and would have linear measurement properties such that a given change in score corresponds to the same amount of change in underlying ability at any part of the ability continuum. Using item response theory methods, linearity of test measurement can be directly assessed and items can be selected to construct a test with desired measurement characteristics. The purpose of this study was to apply item response theory methods to evaluating and developing global functioning scales. Subjects were 1207 patients who had received comprehensive dementia evaluations. Items were selected from two measures of cognitive functioning (Mini Mental State Examination, MMS; Blessed Information Memory Concentration Test, BIMCT) and one measure of independent functioning (Blessed-Roth Dementia Rating Scale, BRDRS). The MMS and BIMCT showed significant non-linearity of measurement, especially at low and high ability levels. A brief composite measure was created by selecting from the three instruments 25 items that fit a uniform distribution of item difficulty across the entire range of ability measured by the three instruments. This composite measure and the BRDRS showed better linearity of measurement than the other two instruments. Results have implications for development of a psychometrically sophisticated, brief measure of global functioning for clinical and research use in dementia. PMID- 10844725 TI - Data management in a longitudinal cross-cultural study. AB - The Indianapolis-Ibadan Dementia Project compares the rates of dementia at two sites, one in the U.S.A. and one in Nigeria. This paper concentrates on the data management issues in this longitudinal cross-cultural study. Approximately 2500 elderly people were recruited at each site, and continue to be re-assessed every two years. All the data are collected on paper and then entered into a FoxPro relational database. Most of the data management, including data cleaning, is done in Indianapolis. The design of the data collection forms is particularly important in a cross-cultural study, with the questions and the coding of responses clear and simple. Since Nigeria and the U.S.A. have different levels of technological development, the computer hardware and software were chosen to be suitable for use at either site. Exchange visits have been needed to address data management issues and resolve unexpected problems. The data management on cross cultural studies can be handled successfully, given careful planning. PMID- 10844726 TI - Modern psychometric methods for detection of differential item functioning: application to cognitive assessment measures. AB - Cognitive screening tests and items have been found to perform differently across groups that differ in terms of education, ethnicity and race. Despite the profound implications that such bias holds for studies in the epidemiology of dementia, little research has been conducted in this area. Using the methods of modern psychometric theory (in addition to those of classical test theory), we examined the performance of the Attention subscale of the Mattis Dementia Rating Scale. Several item response theory models, including the two- and three parameter dichotomous response logistic model, as well as a polytomous response model were compared. (Log-likelihood ratio tests showed that the three-parameter model was not an improvement over the two-parameter model.) Data were collected as part of the ten-study National Institute on Aging Collaborative investigation of special dementia care in institutional settings. The subscale KR-20 estimate for this sample was 0.92. IRT model-based reliability estimates, provided at several points along the latent attribute, ranged from 0.65 to 0.97; the measure was least precise at the less disabled tail of the distribution. Most items performed in similar fashion across education groups; the item characteristic curves were almost identical, indicating little or no differential item functioning (DIF). However, four items were problematic. One item (digit span backwards) demonstrated a large error term in the confirmatory factor analysis; item-fit chi-square statistics developed using BIMAIN confirm this result for the IRT models. Further, the discrimination parameter for that item was low for all education subgroups. Generally, persons with the highest education had a greater probability of passing the item for most levels of theta. Model-based tests of DIF using MULTILOG identified three other items with significant, albeit small, DIF. One item, for example, showed non-uniform DIF in that at the impaired tail of the latent distribution, persons with higher education had a higher probability of correctly responding to the item than did lower education groups, but at less impaired levels, they had a lower probability of a correct response than did lower education groups. Another method of detection identified this item as having DIF (unsigned area statistic=3.05, p<0.01, and 2.96, p<0.01). On average, across the entire score range, the lower education group's probability of answering the item correctly was 0.11 higher than the higher education group's probability. A cross-validation with larger subgroups confirmed the overall result of little DIF for this measure. The methods used for detecting differential item functioning (which may, in turn, be indicative of bias) were applied to a neuropsychological subtest. These methods have been used previously to examine bias in screening measures across education and ethnic and racial subgroups. In addition to the important epidemiological applications of ensuring that screening measures and neuropsychological tests used in diagnoses are free of bias so that more culture-fair classifications will result, these methods are also useful for the examination of site differences in large multi-site clinical trials. It is recommended that these methods receive wider attention in the medical statistical literature. PMID- 10844727 TI - Does an interaction between smoking and drinking influence the risk of Alzheimer's disease? Results from three Canadian data sets. AB - Investigation of the relationship of smoking and drinking to Alzheimer's disease (AD) may advance research on the cause of AD and provide a basis for treatment. Pharmacological mechanisms for an involvement of smoking and drinking are plausible but epidemiologic reports are inconsistent. Evidence of behavioural and physiological interactions suggests that tobacco and alcohol use may not only individually affect AD, but may also modify each other's effects. A modelling strategy was developed to examine the interaction between smoking and drinking on the risk of AD. Three Canadian data sets were analysed: the University of Western Ontario Dementia Study (UWODS) (n=363); the Canadian Study of Health and Aging (CSHA) (n=516), and the database from the Clinic for Alzheimer Disease and Related Disorders at the Vancouver Hospital and Health Sciences Centre, University of British Columbia site (UBC) (n=843). Multiple logistic regression models were adjusted for the potential confounders age, age squared, sex, education, family history of dementia, head injury and hypertension. Analysis of the CSHA provided evidence consistent with the hypothesis that smoking and drinking influence each other's effects on AD, with smoking reducing the risk of AD among drinkers. A similar interaction was marginally significant (p=0.052) in the UWODS data set, but not significant in the UBC data. Extension of these analyses, particularly in longitudinal studies and within genetic risk groups, is needed to determine whether this interaction can be replicated. If so, research on the biological interactions of nicotine and alcohol may provide a basis for the development of therapeutic interventions as well as providing clues to the cause of this disorder. PMID- 10844728 TI - Assessing the relative accuracies of two screening tests in the presence of verification bias. AB - Epidemiological studies of dementia often use two-stage designs because of the relatively low prevalence of the disease and the high cost of ascertaining a diagnosis. The first stage of a two-stage design assesses a large sample with a screening instrument. Then, the subjects are grouped according to their performance on the screening instrument, such as poor, intermediate and good performers. The second stage involves a more extensive diagnostic procedure, such as a clinical assessment, for a particular subset of the study sample selected from each of these groups. However, not all selected subjects have the clinical diagnosis because some subjects may refuse and others are unable to be clinically assessed. Thus, some subjects screened do not have a clinical diagnosis. Furthermore, whether a subject has a clinical diagnosis depends not only on the screening test result but also on other factors, and the sampling fractions for the diagnosis are unknown and have to be estimated. One of the goals in these studies is to assess the relative accuracies of two screening tests. Any analysis using only verified cases may result in verification bias. In this paper, we propose the use of two bootstrap methods to construct confidence intervals for the difference in the accuracies of two screening tests in the presence of verification bias. We illustrate the application of the proposed methods to a simulated data set from a real two-stage study of dementia that has motivated this research. PMID- 10844729 TI - Trace determination of alpha- and beta-endosulfan and three metabolites in human serum by gas chromatography electron capture detection and gas chromatography tandem mass spectrometry. AB - Endosufan, alpha and beta, and three conversion products, sulphate, ether and lactone, were simultaneously determined in human serum by means of an analytical procedure which combines extraction with organic solvents, clean-up with H(2)SO(4) and by liquid column chromatography, and detection by gas chromatography (GC) using electron capture detection (ECD) and tandem mass spectrometry (MS/MS). The procedure was validated and the values of some merit figures, such as linear range, detection and quantitation limits, accuracy, precision and recovery, obtained with the GC/ECD and the GC/MS/MS methods, were compared. The lower limits of detection in GC/ECD and GC/MS/MS were 0.03 and 0.05 microg I(-1), respectively. The recovery of the pesticides at the 20 microg I(-1) concentration level was 60-65%, with the exception of endosufan alpha. Recovery studies at higher levels (100 and 200 microg I(-1)) were independent of pesticide concentration in serum samples. The application of the proposed analytical methodology to the determination of endosulfans and their metabolites in real samples was tested by analyzing serum samples from a population living in agricultural areas of Almeria (Spain). The results show the advantage of MS/MS over the ECD detector in the analysis of serum samples where matrix interferences can be confused with target pesticides. PMID- 10844730 TI - Accurate mass determination by multiple sprayers nano-electrospray mass spectrometry on a magnetic sector instrument. AB - A new technique for accurate mass determination by using multiple sprayers nano electrospray ionization mass spectrometry (nano-ESI-MS) on a magnetic sector instrument is described. Metal coated glass capillaries were used as nano-ESI sprayers. One of the sprayers was used for the reference compound solution, and others were used for the introduction of sample solutions. The spectra of the different compounds were obtained by shifting each sprayer's position relative to the sampling orifice. The accurate masses of several standard compounds were obtained with good accuracy, without problems arising from differences in ionization efficiency between the sample compounds and reference compound. PMID- 10844731 TI - Analysis of single nucleotide polymorphisms by primer extension and matrix assisted laser desorption/ionization time-of-flight mass spectrometry. AB - A method for typing single nucleotide polymorphisms (SNPs) by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) is described, in which a mass-tagged dideoxynucleoside triphosphate is employed in a primer extension reaction in place of an unmodified dideoxynucleoside triphosphate (ddNTP). The increased mass difference due to the presence of the mass-tag greatly facilitates the accurate identification of the added nucleotide, and is particularly useful for typing heterozygous samples. Twenty commercially available mass-tagged dideoxynucleoside triphosphates were screened for amenability to incorporation by AmpliTaq FS and ThermoSequenase DNA polymerases in single nucleotide primer extension (SNuPE) reactions. Several sample preparation and purification methods were also examined and compared. Float dialysis was found to be a simple, versatile, and effective method for purification of the extension products. High specificity and sensitivity were obtained, and all six possible biallelic SNP heterozygotes were determined accurately using a 44-mer synthetic oligonucleotide target DNA as a model system. Further validation of the method was demonstrated in the analysis of five single base mutations in exon IV of the human tyrosinase gene. Single nucleotide variations within 182-bp PCR amplicons amplified from three plasmid and three human genomic DNA samples were genotyped at five variable positions, with results in 100% concordance with conventional sequencing. Genotypes were determined accurately at five sequence-tagged sites (STSs). PMID- 10844732 TI - Perfluoroalkyl ketones: novel derivatization products for the sensitive determination of fatty acids by gas chromatography/mass spectrometry in electron impact and negative chemical ionization modes. AB - Analytically useful pentafluoro ketone derivatives of fatty acids are described. The gas chromatographic/mass spectrometric characteristics of these new derivatives are compared with those of methyl, trimethylsilyl and pentafluorobenzyl esters. Pentafluoro ketones exhibit excellent chromatographic properties and significantly shorter chromatographic retention times than these other esters. The electron impact mass spectra of these new compounds show informative acylium ions, whose intensity decreases with the degree of unsaturation of the parent fatty acid. The formation of strong and informative fragment ions in negative chemical ionization (CH(4)) mass spectra of pentafluoro ketone derivatives allows the detection and the characterization (length of the chain and number of double bonds) of fatty acids at trace levels (femtomole), even in the case of polyunsaturated compounds. The scope and limitations of this new derivatization technique are also discussed. PMID- 10844733 TI - Liquid chromatography tandem mass spectrometric quantitation of sulfamethazine and its metabolites: direct analysis of swine urine by triple quadrupole and by ion trap mass spectrometry. AB - This work describes a new method for the quantitation of trace amounts of sulfamethazine (SMZ) and its main metabolite, N4-acetylsulfamethazine (Ac-SMZ), in swine urine, using high-performance liquid chromatography (HPLC) tandem mass spectrometric analysis of crude urine after addition of internal standard and simple dilution with water. The aim was to determine whether residues of this sulfamidic drug, normally administered to swine in order to prevent infectious diseases, were present in urine at levels lower than those permitted by regulatory authorities before human consumption (EU Project SMT, contract number CT 96-2092). A 10 microL volume of diluted urine was injected into a very short, narrow-bore chromatographic column (Zorbax SB-C18 2.1 i. d. x30 mm length, 3.5 microm pore size). Elution of the analytes of interest was achieved in less than seven minutes using a rapid gradient (from 20 to 80% methanol in 3 minutes). Either a PE Sciex API 365 triple quadrupole (QqQ), operated in the selected reaction monitoring (SRM) mode, or a Finnigan LCQ ion trap (IT) mass spectrometer, operated in narrow-range product ion scan, was used as the final detector. Electrospray (ESI) was used as the ionization technique. A comparison of the two tandem mass spectrometers was performed by analyzing the same set of test samples, at three concentration levels, on three different days. Linearity of responses of the calibration standards, intra- and inter-assay precision of the samples, specificity and limits of detection were evaluated for both systems. Both the QqQ and the IT instrument was suitable for rapid, sensitive and specific determination of the analytes, although the overall performance of the QqQ was slightly superior in terms of linearity, precision and sensitivity. PMID- 10844734 TI - Effect of residual acrylamide monomer from two-dimensional gels on matrix assisted laser desorption/ionization peptide mass mapping experiments. AB - Residual acrylamide can cause severe suppression of signal intensity during matrix-assisted laser desorption/ionization (MALDI) peptide mass mapping experiments. This suppression phenomenon can compromise the ability to detect low picomole and subpicomolar amounts of peptides extracted from two-dimensional gels. A rapid and simple method that exploits the use of pipette tips incorporating C18 packing materials for the enhancement of MALDI signal intensity is presented. The utility of the method is demonstrated with peptide solutions incorporating residual acrylamide and/or gel monomer components. PMID- 10844736 TI - Aspartame degradation study using electrospray ionization mass spectrometry. AB - Electrospray mass spectrometry was used to simultaneously determine aspartame (APM) and five of its degradation products; aspartic acid, aspartylphenylalanine, 5-benzyl-3,6-dioxo-2-piperazieacetic acid (diketopiperazine), phenylalanine, and phenylalanine methyl ester. Under the ionization conditions used, there was no interfering fragmentation for any of the six compounds, i.e., no fragmentation of the compound being tested into other species also being monitored. A study of APM degradation in solution at various pH's and at various temperatures using this method was performed. PMID- 10844735 TI - Coupling of a Rydberg electron capture ion source with a quadrupole mass filter. AB - The coupling of a Rydberg electron capture ion source with a Nermag R10-10H quadrupole mass filter is described. Details are given of the addition to this instrument of a creation cell for atoms excited in Rydberg states. Within the Nermag ion source, such atoms allow attachment of electrons of well-defined thermal energy. SF(6) was used for optimization of the main experimental parameters (gas pressures and voltages applied to the electrodes). The procedure by which Rydberg electron attachment was confirmed is described. A polychlorobiphenyl compound was used to illustrate the performance of this ionization technique. Ion formation was observed in the absence of fragmentation. PMID- 10844737 TI - Analysis of residual solvents in pharmaceuticals with purge-and-membrane mass spectrometry. AB - A method using purge-and-membrane mass spectrometry (PAM-MS) was developed for the analysis of residual solvents in pharmaceutical products. The method combines dynamic headspace and membrane inlet mass spectrometry. The limits of detection for the compounds studied, benzene, toluene, chloroform, 2-pentene and 2-methyl- and 3-methylpentane, were 0.05-0.1 mg/kg. In quantitative analysis the method showed good linearity (r(2) > 0.998) and acceptable within-day (RSD = 7.9-18%) and between-day (RSD = 6.8-10%) repeatability. The PAM-MS method combined with the custom-made Solver program was compared with a method using purge-and-trap gas chromatography/mass spectrometry (P&T-GC/MS) for identification of residual solvents from authentic samples. The results showed that PAM-MS/Solver provides reliable identification of the main volatile organic compounds (VOCs) in the pharmaceuticals, but VOCs with low concentrations (below 0.5 mg/kg) were better identified by P&T-GC/MS. Other advantages of the PAM-MS method were short analysis times and non-requirement for pre-treatment of samples. PMID- 10844738 TI - Sequencing of anti-thyroxine monoclonal antibody fab fragment by ion trap mass spectrometry. AB - A comprehensive mass spectrometric strategy is described for the sequencing of anti-thyroxine monoclonal antibody Fab region (48 000 Da). After reduction and S carboxymethylation of the Fab, the modified light chain and Fd fragment were separated and subjected to multiple proteolytic digestions. The resulting digests were characterized by on-line microbore liquid chromatography/electrospray ionization ion trap mass spectrometry. Database search against published immunoglobulins (IgGs) allowed identification of all the peptides in constant domains. The homologous framework residues in the IgGs were utilized as 'sequence maps' for the sequence determination of variable domains. S-Carboxymethylation with an isotopic-enriched moiety greatly facilitated the recognition and data elucidation of cysteinyl peptides through the unique isotopic distribution patterns specific to the modified peptides. Methylation of peptide mixtures provided additional information for the interpretation of MS/MS spectra, allowing easy differentiation of Asp/Asn and Gln/Glu pairs. This study clearly demonstrates the power of mass spectrometry for the sequencing of antibodies without knowing the corresponding DNA sequences. PMID- 10844739 TI - Sample preparation for high throughput accurate mass analysis by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. AB - An automated sample preparation for high throughput accurate mass determinations by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) has been developed. Sample preparation was performed with an automated workstation and automated mass analyses were performed with a commercial MALDI-TOF mass spectrometer. The method was tested with a 41-sample library. MALDI-TOFMS was found to give the needed sensitivity, accurate mass measurement, and soft ionization necessary for structure confirmation, even of mixtures. A mass accuracy of 5 ppm or less was obtained in over 80% of known compound measurements. A mass accuracy better than 10 ppm was obtained for all measurements of known compounds. Analyses of parallel synthesis products resulted in 77% of the measurements with a mass accuracy of 5 ppm or better. PMID- 10844740 TI - Novel charged [1(4)]azolophanes: associative behaviour revealed by electrospray ionization. AB - Electrospray ionization mass spectrometry has been used to investigate simple multicharged [1(4)]heterophanes containing heterocyclic betaine subunits as building blocks. The formation of stable noncovalent polymolecular self-assembled structures in the gas phase is reported. PMID- 10844741 TI - From holism to integration: is there a future for complementary therapies in the NHS? PMID- 10844742 TI - Feng Shui for the health sector: harmonious buildings, healthier people. PMID- 10844743 TI - Reflexology: recent research approaches. AB - The intention of this paper is to examine recent published research in the field of reflexology and healthcare and consider the way forward for research work in reflexology. It will explore the various research approaches and methods, which have and could be used in answering questions about reflexology practice and outcomes for patients. The recommendations of the Foundation for Integrated Medicine Report (FIM 1997) with regard to conducting research and audit will be included in the paper. These are proposed here as a framework for both reviewing past studies and as guidance for future reflexology investigations. The authors themselves have conducted small reflexology studies and they will reflect on their experiences in analysing the research process. This paper does not aim to give a concise review of the literature to date but uses examples of recently published work to illustrate the issues that might confront researchers. PMID- 10844744 TI - Massage and aromatherapy on a leukaemia unit. PMID- 10844745 TI - The use of massage therapy in palliative care. PMID- 10844746 TI - Aromatherapy and its application in cancer and palliative care. PMID- 10844747 TI - Integrating complementary medicine into the nursing curriculum. AB - As nursing moves into the next century, the changing role of the nurse and increasing scope for practice will create the potential to develop an integrated approach to healthcare and the use of complementary medicine alongside more orthodox care. This paper outlines some of the forces which are creating a change in healthcare integration and discusses the value of pre-registration education and training in helping to make future nursing practitioners understand the use and benefit of complementary medicine. In promoting complementary medicine within the pre-registration curriculum, the process of reflection helped nurse teachers to appreciate the potential to integrate it into nurse training. PMID- 10844748 TI - EMDR: a new treatment for trauma and chronic pain. AB - EMDR (eye movement desensitization and reprocessing) is a new psychological treatment for trauma that is capable of facilitating rapid and permanent reduction in distressing thoughts and feelings (Carlson et al. 1998,Wilson et al. 1995). In addition to reduction of psychological distress, the method leads to more adaptive attitudes and functioning. The utility of the method also appears to extend beyond trauma with positive results reported in the treatment of addictions, phobias, and pain (Henry 1996, Goldstein & Feske 1994, Grant 1986). As a treatment for pain EMDR offers a method of facilitating permanent changes in how pain is experienced somatically and emotionally. Knowledge and understanding of the principles underlying EMDR can also provide a guide for more effective interventions by pain specialists. PMID- 10844749 TI - The Holistic Nurses Association of New South Wales: our history, our present and our future. AB - The Holistic Nurses Association (HNA) of New South Wales was formed in March 1995 by nurses inspired by the inaugural Natural Therapies Conference held by the New South Wales Nurses Association (NSWNA) in October 1994. A philosophy of providing support to nurses wishing to provide healthcare in a holistic way underpinned the formation of the HNA in a climate of disenchantment with the mechanistic and reductionist methods of modern healthcare. The main aims of the group are to share information, meet regularly for support, identify our goals and to organize presentations and workshops that promote and strengthen our professional goals and identity. The HNA has been recognised as a group to consult with regarding policy formation for the use of natural therapies within the mainstream health service. The Nurses Registration Board of New South Wales and the New South Wales Nurses Association both liaised with HNANSW when formulating their policies on the use of complementary therapies in nursing. PMID- 10844750 TI - Complementary medicine: legal status of the non-licensed provider in the USA. AB - European providers of complementary medicine may be surprised by the formal structures which their US counterparts face. A major hurdle to holistic practice in the USA is the statutory prohibition against the unlicensed practice of medicine. State statutes define 'medicine' broadly, creating a legal risk for unlicensed providers of holistic healthcare, as well as for licensed providers whose services are deemed to exceed their legislatively authorized scope of practice and cross into 'diagnosis' and 'treatment' of disease. This paper does not address scope of practice; rather, it focuses on the legal status of non licensed (or 'unenfranchised') providers. For these providers, seeking occupational licensure will provide some protection against medical practice acts, as well as opportunity to elevate professional competence and prestige. However, many holistic providers prefer to remain outside the regulatory scheme. Mandatory licensure, title licensure and registration offer means of upgrading professional status and achieving state sanction for professional practice. PMID- 10844751 TI - Giant olfactory groove meningiomas: advantages of the bilateral fronto orbitonasal approach. AB - Olfactory groove meningiomas are usually considered surgically curable, but the operation becomes complex when the tumour reaches massive size and involves vital suprasellar structures, extends into the ethmoid sinuses or extensively involves the bony floor of the anterior fossa. The traditional bifrontal approach provides only limited exposure in these situations, and a bilateral fronto-orbitonasal approach may be more effective. We reviewed our experience in five patients with resections of massive olfactory groove meningiomas to assess the advantages and risks of these two approaches. All patients were women (mean age, 45 years; range, 28-59) presenting with symptoms and signs related to increased intracranial pressure, cranial nerve impairment or brain compression. Computed tomography, magnetic resonance imaging and cerebral angiography were used for diagnosis and operative planning. The approach was bifrontal in two patients and bifronto-orbitonasal in three. Total surgical removal (Simpson grade I, three patients; Simpson grade II, one patient) was achieved in four patients (80%) and subtotal tumour removal (Simpson grade III) was performed in one (20%). One patient had two staged operations. No morbidity or mortality occurred in this series. No clinical or radiologic evidence of recurrence developed in the four patients whose tumour was resected totally. The bilateral fronto-orbitonasal approach resulted in safe and successful total removal of giant olfactory groove meningiomas, achieving long-term prevention of recurrence. We highly recommend this approach. Copyright 1999 Harcourt Publishers Ltd. PMID- 10844752 TI - Brain TXA(2)and PGI(2)levels related to diffuse brain injury with secondary insults. AB - Secondary insults such as hypotension and hypoxia with head injury are associated with increased mortality and morbidity in comparison with head injury alone. In the present study the changes of brain thromboxane A(2)(TXA(2)) and prostacyclin (PGI(2)) levels in a Marmarou's rodent model of diffuse brain injury with hypotension and hypoxia were observed and the effect of diaspirin cross linked haemoglobin solution (DCLHb) were also investigated. Thirty-two male Sprague Dawley (SD) rats were randomized into four groups: sham; head injury alone; head injury with secondary insults; and injury with insults followed by DCLHb administration. The results showed that there were no changes in TXB(2)and 6-keto PFG(1a)(metabolites of TXA(2)and PGI(2)) levels in the injury alone group, while TXB(2)levels in the secondary insults group were elevated significantly; both TXB(2)and 6-keto-PGF(1a)levels in the injury with insults followed by DCLHb administration group were augmented significantly in comparison with the corresponding value of sham at 4 postimpact. The only increase in TXA(2)levels in the secondary insults rats suggests that an imbalance in TXA(2)-PGI(2)production contributes to the traumatic secondary processes, which include ischaemia and oedema. It is hypothesized that DCLHb may exert its protective properties through increasing PGI(2)production in injured brain by improving blood supply to injured blood vessels. Copyright 1999 Harcourt Publishers Ltd. PMID- 10844753 TI - Absence of demonstrable immune dysregulation in motor neuron disease. AB - Patients with motor neuron disease (MND) (n =33) free of infection were assessed for symptoms of autoimmunity and evidence of peripheral immune activation. Clinical features of autoimmune disease and antinuclear antibodies were notably absent in all patients. Compared with healthy controls, patients with MND had no significant difference in lymphocyte subsets (CD4/8 T cells, B and NK cells). Serum and whole blood endotoxin and phytohaemagglutinin stimulated levels of interferon gamma, granulocyte macrophage colony stimulating factor, interleukin (IL) 1beta, IL4, IL10, tumour necrosis factor alpha and soluble CD23 were no different between patients with MND and healthy controls. Our results confirm the absence of significant elevations of immune regulatory proteins in the peripheral circulation of patients with MND. Copyright 1999 Harcourt Publishers Ltd. PMID- 10844754 TI - Outcome assessment in seizure surgery: the role of postoperative adjustment. AB - The usual criteria employed in assessing the outcome of seizure surgery is almost always that of success in abolition of seizures. Whilst this is a logical method of assessment, experience with large series of such patients shows that this is by no means the only significant criterion. Here we examine the process of postoperative adjustment and the role it plays in determining the outcome of temporal lobectomy for refractory epilepsy. The specific domains involved in this process are examined and it is shown that in a number of cases what is thought to have been surgical success has in fact been significantly diminished by difficulties encountered in postoperative adjustment - to such an extent that total seizure abolition has not resulted in clinical advantage. Copyright 1999 Harcourt Publishers Ltd. PMID- 10844755 TI - Intraventricular neurocytoma: a clinicopathological study of 20 cases with review of the literature. AB - The clinicopathological features of 20 cases of central neurocytomas are described. They accounted for 0.28% of all intracranial tumours diagnosed during a 16 year period (1980-1995). Lower mean age of the patients at diagnosis (23.1 years), male preponderance (M:F=1.8:1) and higher incidence of involvement of the right lateral ventricle (10/20 cases) were noted in this series, in contrast to reports from Western literature. Total removal of the tumour was done in 14 cases while the remaining six underwent partial resection. Morphogically, the tumours had a striking resemblance to oligodendrogliomas (11/20 had been earlier diagnosed as oligodendrogliomas) and an interesting finding was the presence of dilated vascular channels in 12/20 tumours. The diagnosis was confirmed in all cases by immunohistochemistry and/or electron microscopy. While 18 cases were histologically benign, two had features of atypical neurocytoma. Five patients died due to postoperative complications. The remaining patients received postoperative radiation and their follow-up revealed that all of them were doing well at 12 to 72 months after surgery. These neoplasms should be suspected in any young patient with radiological evidence of an intraventricular lesion; for their differentiation from gliomas, immunohistochemistry and electron microscopy should be done. This is important because, unlike gliomas, these tumours have a relatively favourable prognosis and their current treatment of choice is complete surgical removal without adjuvant chemo- or radiotherapy. Copyright 1999 Harcourt Publishers Ltd. PMID- 10844756 TI - Ultra early transient brain swelling following brief intraoperative ischaemia reperfusion. AB - We describe a 50-year-old patient who developed acute transient hemispheric cerebral swelling, documented by computed tomography, following a temporary constriction (20 min) of the middle cerebral artery branches due to clipping of an aneurysm. Reporting of this postischaemic phenomenon has been limited and its mechanism and treatment remain unclear. A possible underlying process could be a postischaemic reactive vasodilatation leading to congestive hyperaemia. Hyperventilation and mannitol seem to be an effective treatment of this essentially 'benign' cerebral swelling. Copyright 1999 Harcourt Publishers Ltd. PMID- 10844757 TI - Bromocriptine-induced apoptosis in pituitary adenoma cells: relationship to p53 and bcl-2 expression. AB - In an attempt to understand the roles of the tumour suppressor gene p53 and the proto-oncogene bcl-2 in cell death and survival in pituitary adenomas, we investigated the relationship of their expression to the apoptotic response of two pituitary adenoma cell lines (GH3 and AtT-20) to bromocriptine. An MTT (3-4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrasolium bromide) assay was performed after treatment with bromocriptine for various periods of time over a range of concentrations to determine the effect of this drug on cell growth. Bromocriptine inhibited growth of GH3 and AtT-20 cells in a dose dependent manner. DNA fragmentation was assessed in GH3 and AtT-20 cells exposed to 10 ug/ml bromocriptine- for 48 h and 72 h. The DNA of GH3 and AtT-20 cells showed nucleosomal fragmentation, indicative of apoptosis. When assayed 2 days after adding bromocriptine, approximately 60% of GH3 and 58% of AtT-20 cells treated with bromocriptine displayed typical apoptotic morphology, including condensed chromatin and fragmented nuclei. There was a time dependent increase in the proportion of all tumour cells undergoing apoptosis. Decreased expression of bcl 2 and accumulation of wild-type p53 were associated with bromocriptine induced apoptosis in pituitary adenoma cells. DNA analysis confirmed the results obtained by the protein study. Different expression of p53 and bcl-2 genes is consistent with the expression of these gene products. These findings show that bromocriptine activated wild-type p53 and suppressed bcl-2 favouring occurrence of apoptosis in pituitary adenoma cells. Copyright 1999 Harcourt Publishers Ltd. PMID- 10844758 TI - Acute effects of an aminosteroid on microdialytically measured parameters after experimental middle cerebral artery occlusion in the rat. AB - The effects of the neuroprotective aminosteroid U74389G were monitored microdialytically in rat cortex during 4 h after permanent middle cerebral artery occlusion. Either U74389G or vehicle only was administered one hour before and 2 h after operation. The dialysate was analysed for on-line pH, ascorbic acid, uric acid, glucose and lactate. In aminosteroid treated animals the levels of ascorbic and uric acids were lower in dialysates after occlusion, total release of lactate was significantly reduced and on-line pH was significantly higher than in control animals. Lesion volume at 4 h, which was significantly reduced in treated animals, correlated positively with ascorbic acid release and on-line pH. Results suggest that neuroprotective effects of aminosteroids might be explained by: (a) preservation of intracellular levels of the radical scavenger ascorbic acid with possible concomitant reduction of glutamate release; and (b) reduced lactate release and increased pH which might influence oedema positively. Copyright 1999 Harcourt Publishers Ltd. PMID- 10844759 TI - The transpetrosal approach for cerebellopontine angle, petroclival and ventral brain stem lesions. AB - Lesions at the cerebellopontine angle, the petroclival region and the ventral aspect of the brain stem are difficult to reach because of their deep seated location and the proximity and complexity of the functionally important surrounding anatomy. Resection of a portion of the petrous bone can provide a wide and direct exposure to these lesions. We employed this technique in the surgery for 39 patients harbouring these lesions which included 21 cerebellopontine angle tumours, nine petroclival tumours and nine ventral brain stem lesions. There was no surgical mortality and the most common morbities were facial nerve palsy (15%), lower cranial nerve palsy (15%) and trigeminal nerve dysfunction (10%). Total removal of tumour or vascular lesion was achieved in 33 patients (85%). Our results indicate that the transpetrosal approach can provide a satisfactory exposure of the cerebellopontine angle, the petroclival region and the ventral brain stem, minimizing the risk of surgical mortality and morbidity. Copyright 1999 Harcourt Publishers Ltd. PMID- 10844760 TI - The zygomaticotemporal approach with medial petrosectomy for intradural lesions. AB - The zygomaticotemporal approach with medial petrosectomy offers an alternative exposure for intradural lesions anterior to the brain stem. It improves access for tumours of the clivus and facilitates proximal control in low lying and giant basilar artery aneurysms. Twenty-three patients were operated on using this approach (12 aneurysms of the posterior circulation and 11 tumours). The permanent complications experienced with this exposure were ipsilateral hearing loss (two cases) and facial nerve palsy (one case). The experience in this series shows it is a useful alternative in accessing this difficult area. Copyright 1999 Harcourt Publishers Ltd. PMID- 10844761 TI - Solitary thoracic intradural extramedullary ependymoma. AB - Intradural extramedullary ependymomas of the cervical or thoracic spine are extremely rare. We present a 62-year-old woman with progressive thoracic back pain over 9 months. Magnetic resonance imaging rerevealed a thoracic intradural extamedullary tumour which was surgically removed in total. Histologic examination identified the tumour as ependymoma, WHO Grade II. Our radiological and surgical findings are consistent with the hypothesis that these tumours develop from heterotopic glial tissue pinched off from the neural tube during its closure. Copyright 1999 Harcourt Publishers Ltd. PMID- 10844762 TI - Magnetic resonance imaging of intradural lumbar disc herniation. AB - The case of a 62-year-old patient with intradural lumbar disc herniation diagnosed by magnetic resonance imaging (MRI) and verified by surgery is reported. The patient presented with low back pain. T(1)-weighted MRI demonstrated an intradural iso-intensity mass at the L2/3 level without obliteration of the extradural fat space. Gd-DTPA enhanced MRI showed a beak like mass with rim enhancement except at its ventral surface. Intraoperative findings disclosed an intradural epiarachnoid fragmented disc which was covered by venous plexus. Copyright 1999 Harcourt Publishers Ltd. PMID- 10844763 TI - Ventriculoperitoneal shunt related meningioma following excision and radiotherapy for glioma. AB - A 25-year-old man with a history of childhood cerebellar glioma treated by surgical removal, radiotherapy and a ventriculoperitoneal (VP) shunt presented with a 5 month history of frontal headaches. Imaging revealed a mass surrounding his shunt, which was surgically removed, with the shunt left in situ. Histopathological examination revealed an atypical and invasive meningioma. A similar case of a meningioma related to a shunt appears in the literature, again in the context of radiotherapy following removal of a previous neoplasm. Copyright 1999 Harcourt Publishers Ltd. PMID- 10844764 TI - An unusual cause of hemifacial spasm. AB - Hemifacial spasm (HFS) is a movement disorder characterized by involuntary paroxysmal chronic contractions of the facial musculature. The usual cause is vascular compression of the seventh cranial nerve, at its exit zone from the brain stem. We report a case of left hemifacial spasm, in a 66-year-old woman, in which the neuroradiological investigation with magnetic resonance imaging showed a hypervascular soft tissue mass arising from the left skull base, in the jugular foramen. This lesion was thought highly likely to be a glomus jugulare tumour. While the usual occurrence of tumour compression causing HFS has been previously recognized, the association of glomus jugulare tumours presenting with HFS has not. The importance of this association is discussed. Copyright 1999 Harcourt Publishers Ltd. PMID- 10844765 TI - Pituitary abscess. AB - A 28-year-old Caucasian woman presented with a 12 month history of secondary amenorrhoea, polyuria and polydipsia with fatigue and weight loss. Investigations revealed panhypopituitarism, diabetes insipidus, an intrasellar mass and papilloedema, thought to be due to benign intracranial hypertension. She was treated conservatively. However, a repeat magnetic resonance image showed enlargement of the pituitary mass with compression of the optic nerves. The pituitary abscess was drained by a transsphenoidal approach. Postoperatively the patient received antibiotics with no recurrence of the pituitary abscess. Copyright 1999 Harcourt Publishers Ltd. PMID- 10844766 TI - Monomelic amyotrophy: non progressive atrophy of the upper limb. AB - Monomelic amyotrophy is a rare clinical entity, resulting in wasting and weakness localized to the hand and forearm unilaterally, in the absence of any sensory or long tract signs. The onset of the disease is insidious, occurring in males before the age of 30 years, with a clinical course marked by non-progression. The case of a 19-year-old Indonesian male patient is presented, with a one year history of right upper limb weakness. Nerve conduction studies were normal, without evidence of conduction block. Electromyography showed changes of chronic partial denervation. Magnetic resonance imaging scans revealed an asymmetry of the spinal cord. Possible aetiological mechanisms for these changes are discussed. Copyright 1999 Harcourt Publishers Ltd. PMID- 10844767 TI - Solitary intradural fibrochondroma in a 16-year-old boy. AB - The following case report is that of a 16-year-old boy who had a solitary benign intracranial parietal fibrochondroma. He presented with a 2 month history of headache. A cranial CT demonstrated an isodense, non-contrast enhancing, extra axial lesion with decreased central opacity and flecks of hyperdensity consistent with calcification. MRI revealed a heterogeneous well circumscribed mass which had an increased signal on T2-weighted images; there was no enhancement with gadolinium. This lesion was totally removed and had histopathological features of a benign chondroma with a fibrocartilaginous character. The literature of this rare tumour is summarized. Copyright 1999 Harcourt Publishers Ltd. PMID- 10844768 TI - Combined peripheral nerve and central nervous system demyelination in a patient with chronic inflammatory demyelinating polyneuropathy. AB - We report the case of a 49-year-old woman suffering combined central and peripheral demyelination, with serial magnetic resonance images scans illustrating the development of central demyelination in a patient with established chronic inflammatory demyelinating polyneuropathy. This is the sixth patient with this rare overlap syndrome reported to date. We review the available literature and argue that an inherited predisposition to both diseases is the most likely explanation for its occurrence. Copyright 1999 Harcourt Publishers Ltd. PMID- 10844769 TI - Craniotomy: the first case histories. AB - The oldest existing case histories of craniotomy are from the false Hippocratic writings, about 330 BC, and one is reconstructed about the death of Ptolemy VI in 145 BC. Greek surgeons had rational indications for trepanning, when the difficulties of the times are understood. All compound fractures were infected, so death from an extradural abscess was likely. Trepanning was intended to drain the extra dural space. Copyright 1999 Harcourt Publishers Ltd. PMID- 10844772 TI - Recent progress in the diagnosis and treatment of multiple sclerosis. AB - Magnetic resonance imaging (MRI) now provides valuable diagnostic and prognostic information for the management of multiple sclerosis (MS) but the diagnosis still largely rests on the clinical features of central nervous system (CNS) lesions disseminated in time and place. Recent histological and MRI studies indicate that extensive axonal damage can occur in MS, even early in the disease course, and is likely to be an important cause of accumulating disability. Several immunomodulating agents have now been shown to have beneficial effects in MS. High dose intravenous or high dose oral methylprednisolone therapy accelerates recovery from attacks of relapsing-remitting MS, but at present there is no convincing evidence that standard dose (intermediate dose) oral corticosteroid therapy is beneficial for such attacks. Interferon beta, copolymer 1 (glatiramer acetate) and i.v. immunoglobulin therapy each significantly reduce the frequency of attacks of relapsing-remitting MS. Interferon beta also inhibits the progression of disability in relapsing-remitting MS and secondary progressive MS, but its effect on primary progressive MS is unknown. Oral low dose methotrexate therapy slows the progression of disability in secondary progressive MS and possibly in primary progressive MS, but it is likely that the currently used dosage (7.5 mg weekly) is suboptimal. Further research is needed to determine the optimal doses and combinations of the above therapies in MS and to develop better therapies, particularly for primary progressive MS. Copyright 1999 Harcourt Publishers Ltd. PMID- 10844773 TI - Clinical and molecular findings in multiple sclerosis patients with type 1 diabetes mellitus. AB - The clinical and molecular findings in three patients with multiple sclerosis (MS) and additional type 1 diabetes mellitus are described. These patients all presented with a severe and progressive disease course of MS. Molecular testing for HLA class II genes demonstrated the presence of the haplotype DRB1*0401, DQB1*0302 in all patients. This haplotype is closely linked to type 1 diabetes mellitus and is increased among patients with the primary progressive subtype of MS. We conclude that the immunogenetic background in patients with diabetes mellitus may determine the severity and clinical course of MS in these individuals. Copyright 1999 Harcourt Publishers Ltd. PMID- 10844774 TI - The effect of optic nerve sheath decompression on CSF dynamics in pseudotumour cerebri and related conditions. AB - The effect of optic nerve sheath decompression (ONSD) on cerebrospinal fluid (CSF) pressure and outflow resistance was studied in six patients who underwent the procedure (four unilateral, two bilateral) for papilloedema secondary to raised intracranial pressure. Four patients (all female, average age 29.3 years) had pseudotumour cerebri while two patients (one male 34 years, one female 24 years) had cryptococcal meningitis. All patients had pre- and postoperative CSF pressure measurements. Three patients had a preoperative CSF infusion study and all six had a postoperative study to measure CSF outflow resistance. Five of the six patients had elevated CSF pressure prior to ONSD and in all five the pressure was still significantly elevated after the procedure. In one symptomatic patient with papilloedema the pressure was normal pre- and post-ONSD (this patient was on acetazolamide). All three patients who tested pre-ONSD had an abnormal outflow resistance; in only one case did this become normal after the procedure, although this patient later required shunting for persistent intracranial hypertension. Clinically, only one of the six cases had a substantial relief of symptoms and signs despite persistently high CSF pressure and outflow resistance. The other five patients went on to shunting after intervals ranging from one week to 8 months. The preliminary conclusion was drawn that ONSD does not have any predictable beneficial effect on CSF pressure or outflow resistance. Copyright 1999 Harcourt Publishers Ltd. PMID- 10844775 TI - Regional differences in cerebrovascular reactivity to acetazolamide in Alzheimer's disease. AB - The cerebral blood flow and cerebrovascular reactivity to acetazolamide were investigated in Alzheimer's disease using stable xenon computed tomography (CT). Ten patients with Alzheimer's disease and 10 healthy controls were studied. The regional cerebral blood flow was measured using the xenon delivery and analysis system (AZ-7000 model, Anzai Sogyo, Tokyo, Japan) and CT (PreSage, Yokogawa Medical Systems, Tokyo, Japan). The subjects inhaled room air followed by a mixture of 30% xenon and 50% oxygen for 3 min. Serial scanning was performed once before xenon inhalation, three times in the wash-in process and five times in the washout process of 5 min. The xenon concentration in the end-tidal expired gas was recorded continuously by the thermoconductivity method. The regional cerebral blood flow was measured before and 20 min after i.v. injection of 17 mg/kg acetazolamide. The blood flows in the frontal lobe, parietal lobe and temporal lobe were reduced but the blood flows in the caudate nucleus, putamen and thalamus were normal in the Alzheimer's disease group. The cerebrovascular reactivity to acetazolamide was reduced in the frontal, parietal and temporal cortex but was normal in the other areas. There is a regional difference in the cerebral blood flow and the cerebrovascular reactivity to acetazolamide in Alzheimer's disease Copyright 1999 Harcourt Publishers Ltd. PMID- 10844776 TI - Treatment of acute relapses in multiple sclerosis at home with oral dexamethasone: a pilot study. AB - The objective of this study was to investigate the feasibility of treating relapses of multiple sclerosis (MS) at home with oral dexamethasone. Twenty-five out of 28 consecutive patients with MS who presented with a relapse of less than 2 weeks' duration were treated on an open basis with oral dexamethasone 16 mg per day (four divided doses) for 5 consecutive days. After one week, the expanded disability status scale (EDSS) had improved by one or more grades in 88% (22 patients) and after 4 weeks in 92% (23 patients). Treatment was well tolerated. We conclude that a course of oral dexamethasone 16 mg per day shortens the duration of an exacerbation in MS in a similar way as seen after high dose i.v. methylprednisolone. Although a randomized study is needed to test this treatment regimen against i.v. high dose corticosteroids, oral dexamethasone can be used in situations when i.v. therapy is difficult to apply. Copyright 1999 Harcourt Publishers Ltd. PMID- 10844777 TI - Transsphenoidal reoperations for removal of pituitary adenomas: rhinological management and timing of reoperation. AB - Transsphenoidal reoperation of pituitary adenomas remain a difficult surgical procedure even with skilful microsurgical techniques. This paper retrospectively analysed how to overcome such difficult factors as absent normal surgical landmarks and an adhesive field. For residual or recurrent adenomas, transsphenoidal reoperations were carried out 14 times in 12 patients. Reasons for reoperations were intentionally multistaged operations in 10 cases, regrowth in three and apoplexy in one. In the first operation, we tried to preserve or restore as many anatomic landmarks as possible. Overly aggressive procedures causing cerebrospinal fluid leakage were avoided. In the three second reoperations within 3 months, a transnasal approach was used. In the other eight situations, between 4 and 119 months, transnasal approach in five, and transseptal approach in three, were used retrospectively. In two patients with fibrous adenomas, reoperations were carried out 4 and 12 months after the first. Magnetic resonance imaging (MRI) evaluation is essential to confirm the tumour descent toward or into pituitary fossa. Timing of the reoperation needs to be individualized depending on MRI findings. Trauma to the nasal mucosa and a narrower operative field became more frequent in reoperations 4 months or later. To obtain a surgical field wide enough to manipulate an ultrasonic aspirator for fibrous adenomas, removal of bony structures at the nasal cavity on the surgical corridor is useful. These rhinological preparations in the first and subsequent operations along with a selection of timing of reoperation are important to gain easier access for tumour removal and in avoidance of surgical complications. Copyright 1999 Harcourt Publishers Ltd. PMID- 10844778 TI - Intracerebral laser Doppler blood flow measurements compared to blood flow in porcine internal carotid artery. AB - The aim was to validate intracerebral laser Doppler measurements: a technique with potential applications for studying cerebral microcirculation. A technique for measurements of blood flow from the internal carotid artery was developed and compared to the laser Doppler method in a pig model. Cerebral blood flow was varied using haemorrhage, high cerebrospinal fluid pressure and blood volume expansion. The coefficient of correlation between flow in the carotid artery and laser Doppler flow signals from probes in the cerebral cortex was 0.29 (P<0.005). The correlation improved using relative changes from baseline (coefficient of correlation=0.62, P<0.0001). The highest coefficient of correlation (0.85, P<0.0001) was obtained when blood flow in the right internal carotid artery and laser Doppler flow signal from the corresponding right hemisphere were compared. Laser Doppler flowmetry from intracerebral probes correlates with the internal carotid artery blood flow. Laser Doppler flowmetry from intracerebral probes may provide a simple method for continuous local blood flow measurement in the brain. Copyright 1999 Harcourt Publishers Ltd. PMID- 10844779 TI - Fasudil, a protein kinase inhibitor, prevents the development of endothelial injury and neutrophil infiltration in a two-haemorrhage canine subarachnoid model. AB - We examined the possible prophylactic potential of fasudil, a protein kinase inhibitor, on the development of endothelial injury and neutrophil infiltration after subarachnoid haemorrhage (SAH). Using the two haemorrhage canine model, fasudil (3 mg/kg) was infused intravenously for 30 min twice daily (days 1-7) and related histological changes were observed by light and electron microscopy. On day 7 characteristic features of the basilar arteries included corrugation of the elastic lamina and endothelial disruption; fasudil inhibited this endothelial damage. Marked neutrophil infiltration into the subarachnoid space was not detected until day 3. On day 7 a large number of neutrophils was observed in the subarachnoid space around all the basilar arteries examined; fasudil treatment significantly inhibited neutrophil infiltration. Our findings suggest that: (1) endothelial injury and neutrophils play a major role in the pathogenesis of cerebral vasospasm; and (2) fasudil inhibited both endothelial damage and neutrophil infiltration, and therefore protein kinase pathways may have a role in these pathological events. Copyright 1999 Harcourt Publishers Ltd. PMID- 10844780 TI - Microsurgical anatomy of the subtentorial supracerebellar and infracerebellar approach to the trigeminal and facial nerves. AB - The microsurgical anatomy of the subtentorial supracerebellar approach to the trigeminal nerve and the infracerebellar lateral approach to the facial nerve was examined in five formalin-fixed adult cadavers. Particular attention was directed to the arrangement of the blood vessels within the posterior fossa, especially the superior petrosal vein and its tributaries, the superior cerebellar artery (SCA), the anterior inferior cerebellar artery (AICA) and the posterior inferior cerebellar artery (PICA). The subtentorial supracerebellar approach provides satisfactory exposure of both, the lateral segment of the SCA, which is the most common vessel involved in the neurovascular conflict resulting in trigeminal neuralgia and the y-inverted shaped superior petrosal vein. The latter results more often from the convergence of the pontotrigeminal vein and the vein of the cerebellopontine fissure. The relationship between the AICA, the PICA and the vertebral artery to the facial-vestibulocochlear complex was studied by an infracerebellar lateral approach. The subpial course of the facial nerve, where the vascular compression is usually found which results in hemifacial spasm, was also documented. These two approaches, as well as a historical review of the evolution of these surgical techniques, since the first description by Dandy, are discussed. Copyright 1999 Harcourt Publishers Ltd. PMID- 10844781 TI - Angiotrophic large cell lymphoma mimicking multiple sclerosis associated transverse myelitis. AB - Angiotrophic large cell lymphoma (ALCL) is a rare disease characterized by a proliferation of malignant lymphoid cells, usually of B-cell origin, within the lumina of small vessels and frequently involving the central nervous system (CNS). ALCL generally follows a rapidly fatal course and is rarely diagnosed prior to autopsy. Tumour cells are rarely present on examination of peripheral blood, cerebrospinal fluid (CSF) or bone marrow, therefore, an early diagnosis is heavily dependent on recognition of neurological manifestations which are present in over 80% of cases. A pre-mortem diagnosis of ALCL can be made in cases where brain biopsy has been performed. We report a case of a 45-year-old woman demonstrating clinical and laboratory features suggestive of multiple sclerosis associated transverse myelitis (MSA-TM) in whom ALCL was unexpectedly discovered at autopsy. Microscopic examination revealed diffuse CNS vascular involvement by neoplastic cells and prominent myelopathy of the upper thoracic spinal cord associated with ascending and descending tract degeneration of the spinal motor and sensory tracts. Neoplastic cells were immunoreactive for B-cell lymphoid markers. The present case is the first to demonstrate neoplastic B-cell clonality in ALCL using polymerase chain reaction (PCR) to detect abnormal V-D-J immunoglobulin gene rearrangements. ALCL should be considered in the differential diagnosis whenever a transverse myelitis is present in association with global encephalopathy and elevated serum lactate dehydrogenase. The neurological manifestations of ALCL with particular reference to its presentation as MSA-TM is discussed. Copyright 1999 Harcourt Publishers Ltd. PMID- 10844782 TI - Spontaneous CSF rhinorrhoea secondary to a middle cranial fossa defect. AB - Spontaneous cerebrospinal fluid rhinorrhoea is a clinical entity that is difficult to both diagnose and treat. A case in a 56-year-old woman with a defect in the lateral wall of the sphenoid sinus and middle cranial fossa floor is presented. Pathogenesis and management of this rare condition are discussed. Copyright 1999 Harcourt Publishers Ltd. PMID- 10844783 TI - Chronic epidural haematoma: a report of 10 cases and review of the literature. AB - Ten cases of chronic epidural haematoma diagnosed between March 1988 and December 1996 are reported. There were eight males and two females ranging in age from 12 48 years. The interval between head injury and the time of diagnosis ranged from 13 days to 3 months with a mean duration of 28 days. There was no mortality or morbidity in this series. The features of chronic epidural haematoma including patient characteristics, time to diagnosis, incidents, pathophysiology, computerized tomographic (CT) scan findings, operative findings, treatment and prognosis are discussed and reviewed in conjunction with the literature. Copyright 1999 Harcourt Publishers Ltd. PMID- 10844784 TI - Polymicrobial brain stem abscess due to Streptococcus anginosus and Actinomyces species. AB - This report describes a rare case of a pyogenic brain stem abscess. The lesion was readily identified by computed tomographic (CT) scan and magnetic resonance imaging (MRI). Streptococcus anginosus was grown from the pus and Actinomyces colonies were found on histopathology. The lesion was successfully treated with stereotactic surgery and antibiotic therapy. A review of the relevant literature including the role of stereotaxy in the treatment of the brain stem abscess is discussed. Copyright 1999 Harcourt Publishers Ltd. PMID- 10844785 TI - Tension pneumocephalus: a case report and literature review. AB - Following a bifrontal craniotomy with frontal sinus involvement, a 64-year-old woman developed cerebrospinal fluid (CSF) rhinorrhoea. At a second operation the skull base dural defect was repaired from within the anterior cranial fossa. Ten days later, the patient developed signs and symptoms of raised intracranial pressure and mass effect. Following radiological investigation, the diagnosis of tension pneumocephalus was made and confirmed at the time of decompression. Within 24 h air had re-accumulated under pressure and concurrently an ongoing CSF leak, which until then had been subclinical, became apparent. Definitive repair of the fistula was performed via a frontal sinus approach and the patient made a total recovery. The relationship between CSF leak and delayed onset tension pneumocephalus is discussed, along with other significant issues regarding the diagnosis and management of this uncommon but dangerous condition. Copyright 1999 Harcourt Publishers Ltd. PMID- 10844786 TI - Internal carotid-posterior communicating artery aneurysm associated with unusually short posterior communicating artery. AB - We report two cases of a ruptured internal carotid-posterior communicating artery aneurysm in which the ipsilateral posterior communicating artery is short and the P1 and P2 segments of the posterior cerebral artery are long. Unique angiographic and operative findings are described. Vertebral angiography with carotid compression is recommended to diagnose this rare variation. Recognition of this variation is important to avoid an inadvertent occlusion of arteries behind the aneurysm at the time of clipping. Copyright 1999 Harcourt Publishers Ltd. PMID- 10844789 TI - Modernity and the emerging futurism in neurosurgery. AB - This article discusses the emergence of neurosurgery in its 'modern' form during the second half of the 20th century and presents the apertures to the 21st century that are apparent in establishing an evolving futurism in the field. Factors of primary positive impetus and challenges are discussed. PMID- 10844790 TI - New antiepileptic drugs. PMID- 10844791 TI - Surgical approaches to mesial temporal structures for epilepsy: a personal view. PMID- 10844792 TI - Postoperative seizure outcome in a series of 114 patients with supratentorial arteriovenous malformations. AB - The incidence of de novo and ongoing postoperative seizures and factors implicated in an increased likelihood of seizures following supratentorial cerebral arteriovenous malformation (AVM) resection remain controversial. We investigated the frequency, severity and variables associated with postoperative seizures in 114 consecutive patients who underwent complete surgical excision of supratentorial AVMs at our institution. The minimal follow up period was 24 months. The incidence of seizures post-AVM surgery was 21% (less than half that found preoperatively). The incidence of postoperative seizures first manifesting >12 months post-AVM resection was 6.3%. A history of preoperative seizures was associated with an increased likelihood of multiple (> or =4) seizures >1 month post-AVM resection (chi2 = 4.38, P = 0.04). Poor functional neurological outcome at 12 months was also a risk factor for the development of > or =1 postoperative seizure using logistic regression analysis (P = 0.04, odds ratio 1.52, 95% CI 1.01-2.28). Cessation of AED therapy in all patients who remain seizure-free at 12 months post-AVM resection is appropriate due to a low risk of new seizure onset or seizure recurrence. PMID- 10844793 TI - Comparative performance of magnetic resonance angiography and conventional angiography in moyamoya disease. AB - Patients with moyamoya disease verified by conventional cerebral angiography (CCA) were evaluated by means of magnetic resonance angiography (MRA), in order to clarify its advantages and limitations in comparison with CCA. This retrospective study was carried out in 13 hemispheres of seven patients with moyamoya disease (including one atypical patient). In MRA, magnetisation transfer contrast (MTC) and maximal intensity projection (MIP) techniques were used. A good correspondence between MRA and CCA was noted in 11 hemispheres (86.4%) on evaluating stenotic lesions and overestimation in the remaining 2. Underestimation of moyamoya vessels in MRA was noted in 3 hemispheres, since well developed moyamoya vessels were detectable, while poorly developed ones were not. No difference between MRA and CCA findings was found in 10 hemispheres (76.9%). MRA tends to overestimate the stenosis and underestimate moyamoya vessels. These characteristics may lead to wrong classification in the angiographic grade of patients with early and advanced stages and should be taken into consideration in interpreting MRA images of moyamoya disease (compatible rate of grade between MRA and CCA; 84.6%). In conclusion MRA with MTC and MIP techniques offers an acceptable quality of assessment of stenotic lesions and moyamoya vessels. MRA is a useful follow up method at present and in the near future it may replace CCA as the initial diagnostic tool. PMID- 10844794 TI - Expression and localisation of urokinase-type plasminogen activator gene in gliomas. AB - Urokinase-type plasminogen activator (uPA) activity is related to the malignant biological behaviour of tumours. In the present study, we examined the presence and distribution of uPA in human gliomas. The amounts of uPA were measured by Northern blotting hybridisation and immunohistochemistry in 43 gliomas and five normal brain specimens. Their relation to the clinical history was comprehensively analysed. All tissues expressed 2.5 kb transcripts of uPA mRNA. The uPA mRNA levels were significantly higher in high grade gliomas than they were in low grade gliomas and normal brain tissue (P < 0.01). Levels of uPA mRNA expression in tumour tissues with recurrence within 18 months postoperatively and survival less than 3 years were significantly higher than counterparts (P < 0.01). The uPA mRNA was also expressed in tumour cells near necrotic areas. The uPA protein expression was consistent with the uPA mRNA expression. The distribution of uPA protein immunoreactivity was mainly in tumour cells and microvascular endothelial cells of glioblastomas and anaplastic astrocytomas, localising in the cytoplasm, especially at sites of vascular proliferation and at the leading edges of tumours. These results suggest that expression of the uPA gene is associated with the malignant progression of gliomas and may play an important role in the recurrence and invasive behaviors of higher grade gliomas. PMID- 10844795 TI - Potentially reversible cognitive impairment in patients presenting to a memory disorders clinic. AB - This paper aims to describe a memory disorders clinic (MDC) for the assessment of patients presenting with early cognitive impairment, to examine the predictive utility of laboratory tests conducted on patients with early memory complaints, and, to investigate the potential reversibility of cognitive impairment in 80 consecutive patients presenting to an MDC. Abnormal laboratory results did not significantly correlate with diagnosis at presentation, with presence or absence of cognitive impairment at presentation or at one year follow up, or with cognitive outcome. Thorough assessment of a person presenting with memory impairment is mandatory. We suggest that a routine battery of laboratory investigations be replaced with selected investigations based on clinical indicators only. The term 'reversible dementia' is a misnomer, and we recommend instead the use of 'potentially reversible cognitive impairment'. We argue that the term 'dementia' should only be used to describe cognitive impairment in cases of irreversible degenerative brain disease. PMID- 10844796 TI - Spinal angulation after anterior discectomy and graftless fusion. AB - A study was made to assess the degree of kyphotic angulation after anterior cervical discectomy and graftless fusion. The possibility of a progressive increase in kyphotic angle was also examined. Fifty-two consecutive patients presenting for review after anterior cervical discectomy and graftless fusion were examined through the use of lateral cervical radiographs and the presence and degree of kyphotic angulation was examined. The majority of operated levels (66%) demonstrated 0-5 degrees of kyphotic angulation and a further 14 (24%) demonstrated between 6 degrees and 10 degrees of kyphotic angulation. PMID- 10844797 TI - Therapeutic strategies and surgical results for pineal region tumours. AB - Until recently, surgery for pineal region tumours has met with poor results. Although experience remains limited, developments in imaging, surgical approaches, and microsurgery have improved outcomes. Over 26 years we treated 36 patients including: 24 with germinoma; four, teratoma; three, pineal cyst; and one each, embryonal carcinoma, choriocarcinoma, pineocytoma, pineoblastoma and metastasis. All 24 germinomas responded to radiotherapy, as did one case each for teratoma, pineoblastoma, and choriocarcinoma. Eight patients underwent resection, and one patient with germinoma had biopsy via endoscopy. Occipital transtentorial and parieto-occipital transcallosal approaches were used in three each, and infratentorial supracerebellar and a combined infratentorial supracerebellar and occipital transtentorial approach in one each. Total removal was achieved in four patients (50%), and subtotal removal in two. Postoperative complications resolved within 2 weeks. Two patients had recurrence and one had dissemination. All tumours except germinomas should be explored and diagnosed histologically. Current operative techniques allow safe, effective removal of pineal region tumours. PMID- 10844798 TI - A comprehensive classification system of vestibular schwannomas. AB - Because traditional classifications of vestibular schwannomas (according to relative size) cannot comprehensively describe lesions that grow in different patterns after arising in regions as diverse as the cerebellopontine (CP) angle, the internal auditory canal, and the region lateral to the fundus of the internal auditory canal (labyrinth), we developed a new system to classify vestibular schwannomas, a system that describes the anatomical structures involved by the tumour, rather than size alone. The vestibular schwannoma is classified first by location and then by extent. Our system provides surgeons information helpful in choosing the surgical approach, in estimating the difficulty of tumour excision, and in determining whether hearing might be preserved. Our system also avoids confusion and misunderstanding in discussions of treatment results because it reflects the diverse biological characteristics of vestibular schwannomas. PMID- 10844799 TI - Alternative tricortical methods of screw implantation for anterior cervical plate fixation: a preliminary report. AB - An experience with 26 cases of anterior cervical interbody plate fixation with an alternative method of screw implantation is described. The follow up ranges from 15 to 51 months. The screws are placed obliquely such that they engage the anterior cortex of the body and traverse through the cortices adjoining the disc space. By this method the screws not only hold the plate firmly with a tricortical purchase, but also, by virtue of their course, stabilise the two adjoining vertebral bodies by themselves. In five of these cases only tricortical screws without the metal plate were used for fixation. Cortico-cancellous iliac crest bone graft shredded into small pieces is placed between the vertebral bodies after adequately preparing the bed. A 100% bony union rate was achieved, with no morbidity or instrument fatigue or failure. The described technique of plate fixation appears to be biomechanically stronger. It provided a rigid segmental internal fixation permitting early mobilisation of the patient with minimal external support. PMID- 10844800 TI - Induction of CD44 expression in stab wounds of the brain: long term persistence of CD44 expression. AB - We studied the expression of the hyaluronan receptor protein CD44 in the mouse brain in response to stab injuries. CD44 expression was strongly activated in the area surrounding the injury within 2 days and then persisted for over 2 months. The expression extended in a direct line the depth of the actual wound inflicted. It appears that CD44 may be involved in the wound healing processes following injury to the brain. PMID- 10844801 TI - Upregulation of amyloid precursor protein and its mRNA in an experimental model of paediatric head injury. AB - Amyloid precursor protein (APP), a membrane spanning glycoprotein which plays an important role in neuronal growth and synaptic plasticity, is increased after traumatic brain injury (TBI) and has been used as a sensitive marker of neuronal damage in an adult sheep head impact model. We hypothesised that APP expression would similarly be increased in lambs, suggesting that in the immature injured brain APP is also upregulated as an acute phase response to trauma. Ten anaesthetised and ventilated 4-5 week old Merino lambs sustained a left temporal head impact from a humane stunner. At 2 h after impact, there was widespread and intense neuronal cell body APP immunoreactivity which was more widely distributed than axonal APP. APP messenger RNA (mRNA) expression was also markedly increased with a distribution similar to that of APP antigen. These results demonstrate that APP antigen and mRNA are sensitive early indicators of TBI in paediatric cases. PMID- 10844802 TI - Growth hormone secreting adenoma with unusual extension: coexisting pituitary cyst and its clinical significance. AB - A 58 year old man showed acromegalic features. The serum growth hormone (GH) level was 7.3 ng/ml and SMC (somatomedin-C) 637 U/ml. Triple stimulation test showed abnormal response compatible with a GH secreting tumour. The conventional enhanced MRI revealed a less enhanced hemisphere-shaped lesion at the right corner of the sella turcica. In addition, dynamic MRI demonstrated an elongated lesion extending to the left beyond the midline. The patient underwent transsphenoidal surgery. Besides the soft and suckable tumour at the right corner, we entered into a small cavity loosely filled with the tumour, which was subsequently also removed. The operative finding corresponded to the lesion shown in dynamic MRI. Postoperative GH and SMC levels became 2.3 ng/ml and 326 U/ml respectively. Incidental pituitary cystic lesions in autopsied cases have been reported to be 6-33%. This case had a GH secreting adenoma with coexisting pituitary cyst. The coexisting pituitary cyst supposedly influenced the unusual shape and extension of the pituitary adenoma. Coexistence of such lesion should be kept in mind for microadenoma on neuroradiological evaluation and on intraoperative inspection surrounding the tumour. PMID- 10844803 TI - Chronic inflammatory demyelinating polyradiculoneuropathy and severe peripheral oedema: a renal explanation. AB - Inflammatory demyelinating neuropathies have been associated with membranous and focal sclerosing glomerulonephritis. Here we describe a 58 year old man with a clinical history, physical examination and laboratory investigations consistent with chronic inflammatory demyelinating polyradiculoneuropathy (CIDP), who also had severe lower limb and sacral oedema resistant to medical therapy. Mild proteinuria was present and a renal biopsy showed features consistent with focal sclerosing glomerulonephritis (FSGN). The patient's weakness and oedema did not respond to i.v. immunoglobulin or plasmapheresis but responded to high dose oral prednisone. The oedema was not explained by immobility, hypoproteinaemia or local factors. The occurrence of the oedema in a person with CIDP and FSGN and its improvement with prednisone, together with improvement in CIDP and FSGN, suggests that it was immune mediated, possibly due to increased capillary permeability. The presence of renal disease in patients with inflammatory demyelinating neuropathies may be more common than currently realised. PMID- 10844804 TI - Multiple meningiomas: case report and review of the literature. AB - A case of a 69 year old woman with multiple meningiomas is presented. Since her first presentation with Jacksonian seizures in 1978, she has had 16 meningiomas excised in seven separate surgical procedures. This is extremely unusual due to both the large number of tumours, and the long period over which they have occurred. We review the epidemiology, possible pathogenesis and genetic associations of this condition. PMID- 10844805 TI - Ocular signs due to an oculomotor intranuclear lesion: palsy of adduction and contralateral eye elevation. AB - Reports on ocular signs of discrete oculomotor nuclear lesions have been rare. This is a case report of a patient with the sudden onset of limited adduction on the left side and bilateral elevation palsy, more pronounced on the right side. The symptoms lasted for 3 days. Neuroimaging study did not reveal a responsible lesion. The patient was diagnosed as having a lacunar infarct. It is neuroanatomically established that the oculomotor subnucleus to the superior rectus muscle primarily cross-innervates the muscle. The palsy of adduction and contralateral supraduction is most plausibly explained by a partial oculomotor nuclear lesion. This patient demonstrated the intranuclear close arrangement of the nerves for the superior and medial rectus muscles. This case reminds us of the clinical importance of basic anatomy based neurological examinations in this computer orientated, high tech era. PMID- 10844806 TI - Rasmussen's syndrome in a 54 year old female: more support for an adult variant. AB - Rasmussen's syndrome, a syndrome of chronic focal encephalitis, is usually considered to be a disease of childhood. Typical features include intractable focal seizures and progressive unilateral neurological deficits with radiological evidence of focal cortical atrophy. This report documents the case of the oldest patient yet described in the literature with Rasmussen's syndrome. Magnetic resonance imaging revealed gadolinium enhancing tissue, not previously described in this condition. PMID- 10844807 TI - Global warming and performance of therapeutic interventions in clinical neurology. PMID- 10844808 TI - Cervical root injury caused by vertebral artery dissection. PMID- 10844811 TI - Heparin should not be used in acute ischaemic stroke. PMID- 10844812 TI - Heparin should not be used presently for acute ischaemic stroke. PMID- 10844815 TI - [Model project for updating neonatal screening in Bavaria: concept and initial results]. AB - The newborn screening programme in Bavaria was confronted with several problems. Number of disorders and process quality no longer complied with screening guidelines. Mixed financing, distributed between the state (PKU, galactosaemia) and health insurances (hypothyroidism) had promoted an increasing dissipation of the system. Notified participation rates had dropped to < 80%. Increasing need for a second screening due to early discharge was an additional challenge. To overcome these problems, and considering the availability of improved screening methodology (tandem mass spectrometry) the programme was reorganised. The project, which started on Jan 1, 1999, is based on a cooperation model between laboratory (logistics, analysis), universities (treatment, scientific evaluation), and public health services (coordination, tracking). Time of blood sampling was predated to the third day of life. Screening was extended to biotinidase deficiency, congenital adrenal hyperplasia (CAH) and by introduction of tandem mass spectrometry for screening of many other disorders (besides PKU). Insurances now finance complete laboratory analysis which was transferred to the private sector. To enable all newborn to participate, the names of screened children are matched against birth lists by public health services on a regional basis. Recalls and conspicuous results are consistently followed up until disorders are either excluded or confirmed. Two clinical hotlines were established in the children's hospitals of the universities in Munich (Southern Bavaria) and in Erlangen (Northern Bavaria). Written consent is required for participation in the programme. Participation in the new programme could be continually increased; coverage is > 95% since April. In several cases screening was made up for not tested children by contacting their parents. Omitted screening was mostly due to misunderstandings regarding testing responsibility or lost samples. Altogether 52 cases of disorder were found in the 87,000 newborn screened until August 1999. Hence, the detection rate of children affected by inborn errors of metabolism was about twice as high than before changes. Among the newly screened diseases CAH was detected most often (11 cases). In 22 cases diagnosis was based on the use of tandem mass spectrometry. Among these (besides PKU, 9 cases) MCAD deficiency (6 cases) was detected most frequently. Whereas recall rates of most disorders were < 0.1%, screening for CAH still revealed a high recall rate, particularly in premature births. Second screening due to early discharge (< 48 h) was required in 1.3%. About 20% of pending recalls required contacting birth hospitals, doctors, midwives or parents. So far all affected children could be brought to treatment in time. PMID- 10844816 TI - [Cost effectiveness of mass orthoptic screening in kindergarten for early detection of developmental vision disorders]. AB - PURPOSE: Orthoptic screening in the kindergarten is one option to improve early detection of amblyopia in children aged 3 years. The purpose of this study was to analyse the cost-effectiveness of such a screening programme in Germany. METHODS: Based on data from the literature and own experience gained from orthoptic screening in kindergarten a decision-analytic model was developed. According to the model, all children in kindergarten, aged 3 years, who had not been treated for amblyopia before, were subjected to an orthoptic examination. Non-cooperative children were reexamined in kindergarten after one year. Children with positive test results were examined by an ophthalmologist for diagnosis. Effects were measured by the number of newly diagnosed cases of amblyopia, non-obvious strabismus and amblyogenic refractive errors. Direct costs were estimated from a third-party payer perspective. The influence of uncertain model parameters was tested by sensitivity analysis. RESULTS: In the base analysis the cost per orthoptic screening test was DM 15.39. Examination by an ophthalmologist cost DM 71.20. The total cost of the screening programme in all German kindergartens was DM 6.1 million. With a 1.5% age-specific prevalence of undiagnosed cases, a sensitivity of 95% and a specificity of 98%, a total of 4,261 new cases would be detected. The cost-effectiveness ratio was DM 1,421 per case detected. Sensitivity analysis showed considerable influence of prevalence and specificity on the cost-effectiveness ratio. It was more cost-effective to re-screen non cooperative children in kindergarten than to have them examined by an ophthalmologist straight-away. CONCLUSIONS: The decision-analytic model showed stable results which may serve as a basis for discussion on the implementation of orthoptic screening and for planning a field study. PMID- 10844817 TI - [Corroboration of concurrent assessment of inappropriate hospitalization with retrospective evaluation based on patient records]. AB - BACKGROUND AND PURPOSE: Since the availability of a German adaptation of the "appropriateness Evaluation Protocol" (AEP) for hospital utilization review, physicians and health care managers were concerned about a potential overestimation of the extent of inappropriate hospital use in a retrospective assessment. The objective was to assess the agreement of concurrent and retrospective assessment of inappropriate hospital use. METHODS: The appropriateness of 42 admissions to surgical wards in a teaching hospital was tested concurrent to the admission. In 25 hospitalized patients the appropriateness of hospital days was assessed. Results were compared with the retrospective testing of the same patients after three months. RESULTS: Agreement between concurrent and retrospective assessment within the same reviewer was 86% of admissions (95% CI = 75%-96%) and 96% of hospital days (88%-100%). Kappa showed values of 0.85 and 0.78, which can be described as excellent. A comparison of two different reviewers, one testing concurrently, the other retrospectively showed agreements of 86% and 90%. The proportion of inappropriate days was the same in both designs. CONCLUSIONS: The results refute prior concerns that a retrospective assessment of the appropriateness of hospital use is impossible because in a retrospective design important clinical information would remain unconsidered or clinical information not yet available at that time would be used for the evaluation. The possibility of an incorrect assessment due to a retrospective design is within the variability inherent to the instrument. Despite the strong agreement, the small sample of surgical patient limits the generalizability of results. PMID- 10844818 TI - [Dementia syndrome and Alzheimer disease: an assessment of morbidity and annual incidence in Germany]. AB - Due to the increasing life expectancy and the growing number of elderly people, dementia is emerging as a major health problem. There is, however, a lack of consistent data on the number of prevalent and incident cases of dementia in Germany. The aim of the present study was to estimate the prevalence and the incidence of dementing disorders in general as well as Alzheimer's disease in particular on the basis of disease rates based on large-scale epidemiological field studies and meta-analyses, which were standardized according to the age structure of the German population. The results show that probably more than 900,000 persons in Germany suffer from a dementing illness, 650,000 among them from Alzheimer's disease. About 200,000 new cases of dementia develop during one year, among them 120,000 new cases of Alzheimer's disease. Due to the higher life expectancy of women as compared to men 70% of the patients are of female gender. As a consequence of the steep increase of disease rates with age, two-thirds of the prevalent cases and 60% of the incident cases are found among people in the age bracket of 80 and over, whereas only 3% are found among persons younger than 65 years of age. Additionally, an estimate of the distribution of cases by the 16 Federal states of Germany is given. PMID- 10844820 TI - [Mortality due to road traffic in Baden-Wurttemberg--air pollution, accidents, noise]. AB - Aim of this study was to determine the influence of road traffic on mortality for the area of Baden-Wurttemberg (Germany) in 1997. Of special interest was the impact of air pollution and road traffic accidents on life expectancy. METHODS: Having the most pronounced effect on mortality particulate matter 10 (PM 10) was chosen for estimation of air pollution. Following the method described by Kunzli et al. PM 10 concentration due to road traffic was estimated from total suspended particles to be 11.1 mg/m3 and relative risk (RR) of PM 10 on mortality be 1.044 (confidence interval 1.033-1.054) per 10 microg/m3 PM 10. RESULTS: 5216 (error range 3040-8892) premature deaths were calculated by life table analysis to be due to traffic. The vast majority (4325) were estimated to be a consequence of PM 10 pollution. The total influence on average life expectancy was 8.8 months (error range 6.0-13.7) with 5.5 months due to PM 10 and 3.3 months due to accidents. The effect of traffic accidents as well as PM 10 on mortality was higher for men than for women. CONCLUSION: Road traffic has a strong influence on mortality. The most important impact of traffic on premature mortality is due to air pollution. While the risk of traffic accidents can, to a certain degree, be controlled individually, prevention of deaths due to air pollution requires general guidelines for emission of deleterious substances. PMID- 10844819 TI - [Tattooing and body piercing--experiences from public health infection surveillance by a public health office]. AB - Tattooing and piercing have become increasingly popular in recent years. Both methods involve several medical risks, including transmission of infectious diseases. There are many reports on wound infections as well as transmission of hepatitis and human immunodeficiency viruses etc. According to these facts special hygiene regulations for tattooing and piercing have been published in Germany. Based on these regulations the public health department of the city of Frankfurt am Main, Germany, carried out special hygiene controls in such studios, one a year. Special tattoo or piercing exhibitions were also controlled. Results are reported here. MATERIAL AND METHOD: Studios for tattoos or piercing were informed about hygiene rules and annually controlled from 1995-1999, using a special check list on cleanliness in the studios, disinfection and sterilisation procedures etc. For permission of tattoo and piercing exhibitions special hygiene orders were made mandatory. RESULTS: During 1995-1997 the absolute number of complaints decreased from 20 to 9, in spite of the increasing number of tattoo studios in Frankfurt am Main (from 6 to 10). This was true also of the tattoo and piercing exhibitions. After 1 year without control visits however, an increase of complaints was to be seen in 1999. DISCUSSION: According to our experience tattooists and piercers are interested in good hygiene practice. But our data showing the worsening hygiene data in one year without control visits also demonstrate the necessity of regular controls by the authorities. According to the reports on infectious complications of tattooing and piercing and according to the data reported here hygienic advice and control is an important task of Public Health services. PMID- 10844821 TI - [Methods for quality assurance of medical examination in epidemiological field studies: the "Study of Health in Pomerania" (SHIP)]. AB - The aims of the population-based, cross-sectional survey "Study of Health in Pomerania" (SHIP) are to describe prevalences and distributions of a broad range of diseases, as well as environmental and behavioural risk factors in the region of Pomerania (North-eastern Germany). The examinations are performed in two study units by different physicians, technicians and interviewers and laboratory analyses in two separate laboratories. In addition, the study will take at least 3 years and approximately 7000 persons will be examined. Because of the design of the study a special concept of quality assurance is indispensable. In the present paper methods are presented for quality assurance and control in anthropometry, ECG, echocardiography, abdominal sonography, carotid ultrasound, sonography of the thyroid gland, blood pressure measurements as well as clinical laboratory analyses. The technical control protocols, the training programmes of the observers and methods to establish observer and method variabilities are presented by means of typical examples. The logistic, personal and technical measures for quality assurance and control require appropriate attention. The methods and procedures presented here are expected to provide an effective instrument to continually monitor the quality of medical examinations in the SHIP study. PMID- 10844822 TI - WebCat: the library catalogue on the world wide web. PMID- 10844823 TI - The Raven Department of Education: the principles of effective curriculum planning. PMID- 10844824 TI - VISTA--visitors in specialist training appointments: the future of college sponsorship. PMID- 10844825 TI - Surgeons under stress: what pastoral care should the college provide? AB - From 15-18 September 1999, the Travelling Surgical Society celebrated its 75th anniversary with a meeting in London for 37 member surgeons, culminating in a dinner at The Royal College of Surgeons of England attended by 140 people. During the dinner, RCS President, Mr Barry Jackson, spoke on behalf of the guests, commenting on the many activities of the TSS, including nine visits to Paris which appeared to lack any social relaxation until he came across a reference from an account in 1938: 'on this occasion we did not attend the Moulin Rouge'. Mr Jackson concluded his speech by presenting the TSS with a copy of a book he had received from the estate of the late Mr R W (Bob) Nevin, one-time Dean of St Thomas's Hospital Medical School and a former President of the TSS, who had signed the fly-leaf. Entitled Truants--The Story of Some who Deserted Medicine Yet Triumphed, the book was written by Lord Moynihan based on the Linacre lecture he delivered in Cambridge in 1936, the year of his death. The President of the TSS expressed warm thanks, emphasized friendship and education as being essential ingredients of travelling societies and sent greetings to the College on its forthcoming bicentenary. PMID- 10844826 TI - Opioids in paediatric cardiac anaesthesia. PMID- 10844827 TI - Off-pump coronary artery surgery and the brain. PMID- 10844828 TI - Qualitative research in health care. PMID- 10844829 TI - Reducing stress responses in the pre-bypass phase of open heart surgery in infants and young children: a comparison of different fentanyl doses. AB - High-dose opioids are advocated for paediatric cardiac surgery to suppress stress responses but they can produce unwanted side effects. There are no data on the dose-dependent effects of opioids on the stress response on which to base rational opioid administration. We conducted a dose ranging study on 40 children less than 4 yr undergoing elective open heart surgery using one of five fentanyl doses: 2, 25, 50, 100 or 150 micrograms kg-1 before surgery. The standardized anaesthetic also included pancuronium and isoflurane. Blood samples were taken at induction, before incision, after sternotomy, immediately before, and at the end of cardiopulmonary bypass. Patients in the 2 micrograms kg-1 group had significant rises in prebypass glucose (P < 0.01), pre- and post-bypass cortisol (P < 0.01), and pre- and post-bypass norepinephrine (P < 0.01). No significant rise occurred in glucose, cortisol and catecholamines in any of the higher dosage groups. Patients in the 2 micrograms kg-1 group had significantly higher mean systolic blood pressure (P < 0.02) and heart rate (P < 0.04). A balanced anaesthetic containing fentanyl 25-50 micrograms kg-1 is sufficient to obtund haemodynamic and stress responses to the pre-bypass phase of surgery. Higher doses of fentanyl (100 and 150 micrograms kg-1) offer little advantage over 50 micrograms kg-1, and can necessitate intervention to prevent hypotension. PMID- 10844830 TI - Inspired oxygen fraction after cardiopulmonary bypass: effects on pulmonary function with regard to endothelin-1 concentrations and venous admixture. AB - Twenty consecutive patients undergoing cardiac surgery with cardiopulmonary bypass (CPB) were allocated at random to group 1 (n = 10, high inspired oxygen fraction (FIO2) after CPB), or group 2 (n = 10, moderate FIO2 after CPB). The effects of each FIO2 on arterial and mixed venous concentrations of endothelin-1 (ET-1) and its precursor, Big ET-1, were measured. Venous admixture was calculated to assess the efficiency of pulmonary gas exchange. Patients whose lungs had been ventilated with a FIO2 of 1.0 (exposure time 70 min) after weaning from the CPB machine had significantly greater arterial and mixed venous Big ET-1 concentrations and venous admixture than patients whose lungs were ventilated with a FIO2 of 0.35. In contrast, ET-1 concentrations in the two groups were not significantly different. A reduction of FIO2 from 1.0 to 0.6 reduced venous admixture without lowering endothelial peptide concentrations. On the first postoperative day all peptide concentrations were similar in the two groups, whereas venous admixture remained non-significantly higher in group 1. A short period of high FIO2 immediately after CPB increases endothelin concentrations and pulmonary venous admixture. PMID- 10844831 TI - Assessment of the adequacy of systemic and regional perfusion after cardiac surgery. AB - Changes in systemic, hepatosplanchnic and femoral blood flow and liver function after cardiac surgery were studied in 17 patients from April to October 1995. Blood flows were measured every 3 h and gastric mucosal PCO2 (by tonometry) every hour from arrival in the intensive care unit until extubation. Cardiac output and systemic oxygen consumption increased from 2.83 (0.68) litres min-1 m-2 to 3.17 (0.57) litres min-1 m-2 and from 126 (18) ml min-1 m-2 to 135 (44) ml min-1 m-2, respectively (mean (SD), P = 0.028 and P = 0.019, respectively, baseline vs 6 h). The fraction of cardiac output distributed to the splanchnic region decreased from 0.25 (0.06) to 0.20 (0.04) (P = 0.004) while splanchnic oxygen extraction increased from 0.43 (0.15) to 0.50 (0.12) (P = 0.019). Femoral blood flow increased from 0.18 (0.07) litres min-1 m-2 to 0.23 (0.09) litres min-1 m-2, (P = 0.006, baseline vs 3 h) but femoral oxygen consumption did not change. Changes in blood flow were not reflected by venous-arterial PCO2 gradients. Initially high glutathione transferase alpha concentrations decreased and indocyanine green extraction was well preserved. We conclude that the predominant increase in peripheral blood flow and the increased oxygen uptake in certain regions of the body may increase the risk of a mismatch between splanchnic perfusion and metabolic demands. This mismatch was not associated with impaired liver function or cellular integrity. PMID- 10844833 TI - Severe carbon monoxide poisoning: outcome after hyperbaric oxygen therapy. AB - This paper reports the outcome after carbon monoxide poisoning in 31 consecutive patients treated with mechanical ventilation and hyperbaric oxygen therapy, compared with another study of mechanically ventilated patients treated with normobaric oxygen. We found 16.1% hospital mortality and 3.8% severe short-term memory loss, compared with 30% hospital mortality and 20% incidence of serious neurological deficit after treatment with normobaric oxygen; outcome was poor in 19.4% and 44.3% of those treated with hyperbaric and normobaric oxygen, respectively (P < 0.05). Cerebral oedema caused three of five deaths despite hyperbaric therapy, occurring at 24-48 h after poisoning. Intracranial pressure monitoring and CT scan of the head before wakening should be considered in any severely poisoned patient. PMID- 10844832 TI - Myocardial consequences of remifentanil in patients with coronary artery disease. AB - Remifentanil may be an alternative to conventional opioids for minimally invasive direct coronary artery bypass surgery because of its extremely short duration of action. The aim of this study was to investigate the effects of remifentanil on myocardial blood flow, metabolism and systemic haemodynamic variables in patients with coronary artery disease. After approval by the local ethics committee, 12 male patients were investigated before elective coronary artery bypass grafting. Systemic haemodynamic variables, myocardial blood flow and metabolism were measured when patients were awake and when they were anaesthetized with high-dose remifentanil (2.0 micrograms kg-1 min-1), or with remifentanil 0.5 microgram kg-1 min-1 combined with propofol (target-controlled infusion aiming at a plasma concentration of 2.0 micrograms ml-1). Myocardial blood flow was measured using a modified Kety-Schmidt technique. High-dose remifentanil anaesthesia significantly reduced cardiac index (CI) (-25%) as a consequence of a decrease in stroke volume index (SVI) (-14%) and heart rate (-13%). Mean arterial pressure (MAP) was 30% lower than that in the awake patient. Myocardial blood flow and myocardial oxygen uptake (MVO2) decreased by 30% and 42%, respectively. In contrast to high-dose remifentanil anaesthesia, systemic vascular resistance index (-14%) during remifentanil/propofol anaesthesia was significantly lower than that in the awake patient. Other haemodynamic variables, and myocardial blood flow and MVO2, did not significantly differ from the high-dose remifentanil period. In conclusion, high-dose remifentanil reduces SVI, heart rate, MAP, myocardial blood flow and MVO2 and its effects do not differ from those of remifentanil/propofol anaesthesia. PMID- 10844834 TI - Dose requirements of infusions of cisatracurium or rocuronium during hypothermic cardiopulmonary bypass. AB - We investigated the influence of mild hypothermic cardiopulmonary bypass (CPB) on the dose requirements of cisatracurium or rocuronium used as a continuous infusion. We studied eight patients given cisatracurium and nine given rocuronium. They were ASA class III and IV and scheduled for elective coronary artery bypass grafting. Neuromuscular transmission was monitored electromyographically. After recovery of T1/T0 to 10%, a cisatracurium infusion or a rocuronium infusion was started at a rate of 1.5 or 10 micrograms kg-1 min 1, respectively, and adjusted to maintain T1/T0 at 15%. Infusion rate and duration were recorded before, during and after CPB in each patient and the mean infusion rates were calculated. One-way ANOVA showed a statistically significant difference between the cisatracurium infusion rates before, during and after CPB: A T1/T0 of 15% could be achieved with a mean infusion rate of 1.1, 0.75 and 0.98 micrograms kg-1 min-1 before, during and after CPB, respectively. There was no significant difference between the rocuronium infusion rates before, during and after CPB. The mean rocuronium infusion rate required to maintain T1/T0 at 15% throughout the procedure was 4.1 micrograms kg-1 min-1. Cisatracurium infusion rates should be halved during CPB. Even after CPB, requirements are reduced. The same tendency occurs with rocuronium, but the changes in infusion rate were not statistically significant. PMID- 10844835 TI - A detailed analysis of theatre training activity in a UK teaching hospital. AB - We examined the placement of anaesthetists in our department over a 2 yr period. Data were collected from an in-theatre system to provide details of caseload and supervision for 34,856 operations. There was wide variation between anaesthetic sub-specialties with overall supervision levels of 35% of cases for senior house officers (SHOs) and 32% for specialist registrars (SpRs). The consultant data showed the size and areas of teaching reserve in the department. We then examined individual logbooks in order to validate our data, and departmental rotas to put these data into perspective with previous attempts to quantify trainee supervision. Supervision data derived from the rota allocations showed that 86% of SHO lists and 62% of SpR lists were scheduled to be supervised. This study has described our training activity and facilitated departmental changes, as well as highlighting the need for great care in interpreting trainee supervision data acquired from different sources, particularly when comparisons are being made. PMID- 10844836 TI - Chemiluminescence underestimates nitric oxide concentration in the presence of potent inhalation anaesthetics. AB - We investigated the effect of potent inhalation anaesthetics on nitric oxide (NO) concentration measured by the chemiluminescence method. We found that the NO concentration was increasingly underestimated with increasing concentrations of halothane, isoflurane, enflurane and sevoflurane (r2 = 0.918-0.997, P < 0.01). Statistical analysis showed that the four inhalation agents at the same concentration produced a similar error in the measured NO concentration. In the presence of a fixed concentration of sevoflurane (5.0%), isoflurane (5.2%), enflurane (4.5%) or halothane (6.1%), the rate of reduction in the measured NO concentration increased in proportion to the NO concentration (r2 = 0.909-0.982, P < 0.01). No direct chemical interaction between the potent inhalation agents and NO was detected by gas chromatography-mass spectrometry. We conclude that NO concentration can be underestimated when measured by the chemiluminescence method in the presence of potent inhalation agents. This underestimation may result from emission absorption and/or the quenching phenomenon, but is not attributable to a chemical reaction between the inhalation agent and NO. PMID- 10844837 TI - Comparison of exogenous surfactant and positive end-expiratory pressure therapies in a model of human breast milk-induced acute lung injury in rabbits. AB - To evaluate surfactant and positive end-expiratory pressure (PEEP) as potential therapies for the acute lung injury after tracheal instillation of 0.8 ml kg-1 human breast milk (HBM) acidified to pH 1.8, adult white rabbits were anaesthetized, tracheostomized, ventilated and randomized to (six rabbits per treatment): (i) no treatment after HBM (control); (ii) exogenous Bovine surfactant (100 mg kg-1) 1 h after HBM; (iii) PEEP-pre (0.5 kPa PEEP) before and after HBM; or (iv) PEEP-post (0.5 kPa PEEP) after HBM. A fifth group of six rabbits received no tracheal instillate (no aspirate). The alveolar to arterial oxygen tension gradient (A-aDO2) and dynamic compliance were measured pre-injury and hourly for 4 h. At post-mortem, the lungs were examined histologically. A aDO2 in all four HBM-injured groups increased to a maximum at 1 h post-injury; A aDO2 then returned towards the baseline in the surfactant and PEEP-post groups, but remained increased in the PEEP-pre and control groups. Dynamic compliance decreased in all four HBM-injured groups. A-aDO2 and compliance were unchanged in the no aspirate group. Bronchoalveolar architecture after surfactant therapy was normal. We conclude that surfactant is a more effective therapy for HBM-induced lung injury than either 0.5 kPa PEEP-post or PEEP-pre injury. PMID- 10844838 TI - Fentanyl reduces infarction but not stunning via delta-opioid receptors and protein kinase C in rats. AB - Langendorff rat hearts were used (i) to examine whether fentanyl reduces stunning, infarction or both, and (ii) to investigate if this protection is mediated by delta-opioid receptors and/or protein kinase C (PKC). In the stunning study, hearts were subjected to global ischaemia (20 min) and reperfusion. This did not produce infarction. Postischaemic mechanical function was measured in hearts treated with or without fentanyl (740 nM). Fentanyl did not affect postischaemic mechanical function. In the infarction study, the left anterior descending coronary artery was occluded for 35 min and infarct size was assessed by triphenyltetrazolium chloride staining. Hearts in the control group exhibited an infarct zone/area at risk (I/R) of 39 (SEM 5)%, whereas the I/R for the fentanyl group was 13 (2)%. When the hearts were treated with a delta-opioid receptor antagonist (naltrindole 1 nM) or a PKC inhibitor (chelerythrine 2 microM), the effect of fentanyl was abolished, with I/R of 37 (1) and 36 (2)% respectively. In our model, we conclude that fentanyl protects against infarction but not against stunning, and that the limitation of ischaemic injury is mediated by both delta-opioid receptors and PKC. PMID- 10844839 TI - Thermoregulation, mild perioperative hypothermia and postanaesthetic shivering. PMID- 10844840 TI - Reduced cerebral embolic signals in beating heart coronary surgery detected by transcranial Doppler ultrasound. AB - Cerebral emboli detected by transcranial Doppler imaging were recorded in 20 patients undergoing multiple-vessel coronary artery bypass surgery, either with or without cardiopulmonary bypass, in a prospective unblinded comparative study. Emboli were recorded continuously from the time of pericardial incision until 10 min after the last aortic instrumentation. The numbers of coronary grafts and of aortic clampings were also documented. Patients undergoing revascularization with cardiopulmonary bypass had more emboli (median 79, range 38-876) per case compared with patients having off-pump surgery (median 3, range 0-18). No clinically detectable neurological deficits were seen in either group. Beating heart surgery is associated with fewer emboli than coronary surgery with cardiopulmonary bypass. Further research is necessary to determine whether a smaller number of emboli alters the incidence of neurological deficit after cardiac surgery. PMID- 10844841 TI - Effects of isoflurane, sevoflurane and propofol anaesthesia on jugular venous oxygen saturation in patients undergoing coronary artery bypass surgery. AB - We investigated the effect of sevoflurane, isoflurane and propofol on jugular venous bulb oxygen saturation (SjO2) in 21 patients undergoing coronary artery bypass graft surgery (CABG) during and after normothermic cardiopulmonary bypass (CPB). Patients received a standardized anaesthetic consisting of fentanyl, midazolam and were then randomly allocated to receive either isoflurane, sevoflurane or propofol for maintenance. SjO2 values were significantly lower than baseline 1 h after CPB in the propofol but not the isoflurane or the sevoflurane groups. Furthermore, SjO2 values were significantly higher during CPB in the isoflurane group (P = 0.0081) and significantly lower 6 h after CPB in the sevoflurane group (P = 0.0447) when compared to the propofol group. We conclude that jugular venous desaturation during and after normothermic CPB is more likely during propofol anaesthesia. PMID- 10844842 TI - Effect of anaesthesia on the cardiac response to intravenous adenosine. AB - The cardiac response to intravenous adenosine 112 micrograms kg-1 was studied in 16 patients scheduled for coronary artery bypass surgery before and during anaesthesia with 1% end-tidal isoflurane and fentanyl 10 micrograms kg-1. Mean time from injection to onset of adenosine-induced PR prolongation was significantly greater during anaesthesia (12.8 (SD 5) vs 9.9 (3) s, P = 0.032). Atrioventricular block (assessed by the total number of non-conducted P waves) was significantly less during anaesthesia (12 vs 27, P = 0.016). We conclude that anaesthesia including 1% isoflurane and fentanyl 10 micrograms kg-1 delays the onset and reduces the magnitude of adenosine-induced atrioventricular block. PMID- 10844843 TI - Carbon dioxide elimination during high-frequency jet ventilation for rigid bronchoscopy. AB - Oxygen saturation and carbon dioxide values should be monitored during high frequency jet ventilation (HFJV). Modern transcutaneous PCO2 (PtcCO2) measurement allows the estimation of ventilation efficiency. We studied how tests of lung function could predict carbon dioxide elimination during HFJV. Lung function tests from 180 adult patients undergoing rigid bronchoscopy were analysed as factors affecting carbon dioxide elimination. The lung function test results showed a significant relationship with the efficiency of carbon dioxide elimination; the greatest impairment of carbon dioxide elimination was found in patients with combined abnormalities of lung function. Further factors associated with difficult carbon dioxide elimination were male gender and elevated body weight. Of the patients investigated, 72% had normal carbon dioxide elimination, whereas in 23% hypercapnia could be avoided only by increasing the driving pressure. The prevalence of abnormal preoperative lung function test results predicts (sensitivity 76%, positive predictive value 27%) impaired carbon dioxide elimination during jet ventilation and rigid bronchoscopy. PMID- 10844844 TI - Cp50 of propofol with and without nitrous oxide 67%. AB - The target concentration of propofol required to prevent response to surgical incision was determined in 60 unpremedicated ASA I or II patients, who breathed either oxygen-enriched air or nitrous oxide 67% in oxygen. Propofol was infused using a target-controlled infusion system incorporating the standard 'Diprifusor' pharmacokinetic model, with the target concentration for each patient decided by up/down sequential allocation. Presence or absence of movement in response to a groin incision was determined by the surgeon. The calculated blood concentration at which 50% of patients responded (Cp50calc), determined by probit analysis, was 6.8 micrograms ml-1 for patients who breathed oxygen-enriched air and 4.9 micrograms ml-1 for those who breathed nitrous oxide 67% in oxygen. PMID- 10844845 TI - Anticoagulant-induced pseudothrombocytopenia in a patient presenting for coronary artery bypass grafting. AB - A 73-yr-old man with severe ischaemic heart disease presented for coronary artery bypass grafting. His preoperative platelet count, obtained from an ethylene diamine tetraacetic acid (EDTA) sampling bottle, was 61 x 10(9) litre-1, but he had no history of bleeding problems. Previous platelet counts demonstrated results ranging from 16 x 10(9) litre-1 to 254 x 10(9) litre-1 with variable degrees of in vitro platelet clumping. Preoperative thrombelastography reflected a normal coagulation profile. The laboratory findings and the absence of a history of haemorrhagic complications suggested a diagnosis of EDTA-dependent pseudothrombocytopenia. We present the perioperative implications of this in vitro phenomenon and methods of detecting the functional and numerical integrity of circulating platelets. PMID- 10844846 TI - Oesophageal perforation following perioperative transoesophageal echocardiography. AB - Transoesophageal echocardiography (TOE) is being used more often by cardiothoracic anaesthetists for the perioperative management of cardiac problems. Reports of iatrogenic oesophageal perforation by instrumentation of the oesophagus are increasing. Although TOE is considered safe, it may be more risky during surgery, because the probe is passed and manipulated in an anaesthetized patient. It may be in place for several hours so the risk of mucosal pressure and thermal damage is increased. Patients on cardiopulmonary bypass are also fully anticoagulated. We describe a case of oesophageal perforation following insertion of the TOE probe in a patient with gross cardiomegaly. Oesophageal distortion by cardiac enlargement may increase the risk of oesophageal perforation. Difficulty in passage of the TOE probe should be regarded with suspicion and withdrawal should be contemplated because the symptoms of oesophageal perforation are often delayed and non-specific. Delay in investigation, diagnosis and treatment will increase morbidity and mortality. PMID- 10844847 TI - Mallory-Weiss tear following cardiac surgery: transoesophageal echoprobe or nasogastric tube? AB - A case of fatal upper gastrointestinal bleeding from a Mallory-Weiss tear after transoesophageal echocardiography during cardiac surgery is reported. After the echo-cardiographic examination, which is considered a safe procedure, a nasogastric tube was inserted which immediately revealed bright red blood. Eventually the patient lost 9 litres of blood. The role of the echo-probe and the nasogastric tube in causing the Mallory-Weiss tear is discussed. Although this case is not conclusive about the mechanism of oesophageal damage, it is suggested that the safety recommendations for transoesophageal echocardiography also apply for instrumentation of the oesophagus with a nasogastric tube after the transoesophageal echocardiographic examination. PMID- 10844848 TI - The LMA 'ProSeal'--a laryngeal mask with an oesophageal vent. AB - We describe a new laryngeal mask airway (LMA) that incorporates a second tube placed lateral to the airway tube and ending at the tip of the mask. The second tube is intended to separate the alimentary and respiratory tracts. It should permit access to or escape of fluids from the stomach and reduce the risks of gastric insufflation and pulmonary aspiration. It can also determine the correct positioning of the mask. A second posterior cuff is fitted to improve the seal. A preliminary crossover comparison with the standard mask in 30 adult female patients showed no differences in insertion, trauma or quality of airway. At 60 cm H2O intracuff pressure, the new LMA gave twice the seal pressure of the standard device (P < 0.0001) and permitted blind insertion of a gastric tube in all cases. It is concluded that the new device merits further study. PMID- 10844849 TI - Evidence-based medicine and the Cochrane Collaboration in anaesthesia. PMID- 10844850 TI - Effect of omitting regular ACE inhibitor medication before cardiac surgery on haemodynamic variables and vasoactive drug requirements. PMID- 10844851 TI - The causes of aspiration pneumonia in mechanically ventilated patients; a possible pathological link with upper airway bacterial colonization. PMID- 10844852 TI - Comparison of low dose epidural with combined spinal-epidural analgesia for labour. PMID- 10844853 TI - Alternatives to succinylcholine at caesarean section. PMID- 10844854 TI - Anaphylaxis to rocuronium. PMID- 10844855 TI - Asystole with propofol and remifentanil. PMID- 10844856 TI - Unintended total spinal anaesthesia with ropivacaine. PMID- 10844857 TI - Protecting anaesthetic tubing from occlusion. PMID- 10844858 TI - Cutting needle biopsy in the diagnosis of clinically suspected non-carcinomatous disease of the lung. AB - Most patients referred for lung biopsy have a focal lesion that is likely to be a carcinoma, and fine needle aspiration is usually sufficient to confirm the diagnosis. When non-carcinomatous disease is suspected, tissue architecture is important and potential diagnostic techniques include percutaneous cutting needle biopsy (CNB). We retrospectively reviewed 37 CNBs performed for clinically suspected non-carcinomatous disease; recording the biopsy result, final diagnosis, radiological nature of the pulmonary abnormality, distance from the pleura of the lesion biopsied and biopsy complications. 9 patients had a single pulmonary nodule/mass; 13 had multiple nodules/masses; 8 had a lobar consolidation/mass; and 7 had multifocal consolidation. The lesion abutted the pleura in 31 cases, lying within 1 cm in the other 6 cases. The minor complication rate was 14%, with no major complications. Specific malignant diagnoses were made in 9 patients, and specific benign in 23, in all of whom clinicoradiological follow-up was concordant. CNB did not yield a specific diagnosis in five patients, including two lymphomas and one case of unsuspected tuberculosis in which the sample was not cultured. The overall accuracy of CNB was 32/37 (86%). CNB is a safe and accurate means of achieving a tissue diagnosis for patients with peripheral pulmonary parenchymal disease thought not to represent carcinoma. PMID- 10844859 TI - Do bullae indicate a predisposition to recurrent pneumothorax? AB - A relationship between idiopathic spontaneous pneumothorax (ISP) and visible parenchymal bullae and blebs has been reported. The causal relationship between blebs and bullae and ISP is questionable. Consequently, resection of the involved area is only indicated if it predisposes to recurrence of the pneumothorax. CT studies on 101 cases of ISP were analysed. The presence of bullae and their distribution was then related to the first and recurrent pneumothorax. CT demonstrated bullae in 56% of first ISP and in 64% of recurrent ISP, mostly among older patients. Only eight patients had bullae larger than 2 cm on the side of the pneumothorax. The location of the bullae was not a factor in predicting recurrent pneumothorax. PMID- 10844860 TI - Congenital diaphragmatic herniation: antenatal detection and outcome. AB - 201 fetuses and babies with a congenital diaphragmatic herniation or eventration (referred to collectively as CDH) were notified to the Northern Region Congenital Abnormality Survey (NorCAS) in the 13-year period from 1985 to 1997, an incidence of 0.25 per 1000 births. The 1-year survival of all pregnancies associated with CDH was 37%. The 1-year survival of livebirths was 50%. Antenatal scan detected the diaphragmatic defect or associated structural abnormality in 50%. Another major structural abnormality was present in 62 (31%); one of these babies survived and 26 pregnancies were terminated. This group contained four of the six antepartum stillbirths, all three intrapartum stillbirths and four of the five spontaneous miscarriages. Non-isolated CDH occurred in association with Fryns (5), Goldenhar (1) and de Lange (1) syndromes, and in 16 of 17 with a chromosome anomaly. 53% of the 139 fetuses with isolated CDH survived to 1 year of age, and 59% of the 124 liveborn survived. Of 37 fetuses with isolated CDH detected before 25 weeks gestation, 12 pregnancies were terminated. There were 11 survivors among the 25 continuing pregnancies (44%). The overall survival of babies with CDH is very poor but, when a defect is identified by ultrasound before 25 weeks gestation, chromosome analysis and a careful ultrasound scan may suggest which fetuses have an isolated diaphragmatic abnormality and a greater chance of survival. PMID- 10844861 TI - The chest radiograph appearances of very premature infants at 36 weeks post conceptional age. AB - The chest radiograph of very premature infants at 36 weeks post-conceptional age (PCA) was evaluated with regard to the degree of hyperinflation and cardiomegaly, and the presence of fibrosis/interstitial shadowing, cystic elements, air bronchograms and opacification. The evolution of abnormalities was assessed by comparing the radiograph appearance at 36 weeks PCA with that at 28 days post natal age (PNA). Three scoring systems were used to determine how any abnormalities present could be best quantified to reflect disease severity as determined by chronic dependency upon supplementary oxygen status. Chest radiographs at 36 weeks PCA from 60 infants (median gestational age 26 weeks (range 24-28)) were studied. 47 infants also had radiographs at 28 days PNA. Only three infants had no chest radiograph abnormalities at 36 weeks PCA, although 24 infants were not dependent upon supplementary oxygen. The most common abnormalities were interstitial shadowing and hyperinflation, while cystic elements and cardiomegaly were rare. The radiographic appearance had deteriorated from 28 days PNA to 36 weeks PCA (p < 0.05); more infants at 36 weeks PCA were hyperinflated (p < 0.01). The chest radiograph appearances of infants who were dependent upon supplementary oxygen scored higher than those who were not (p < 0.01) using all three scoring systems. The system that assessed only the presence of interstitial shadowing, cystic elements and hyperinflation had the highest specificity in identifying oxygen dependency beyond 36 weeks PCA and had the highest area under the respective receiver operator characteristic curve. In conclusion, the majority of very immature infants have an abnormal chest radiograph appearance at 36 weeks PCA. The appearance can, however, be meaningfully scored by evaluating only three abnormalities. PMID- 10844862 TI - Skin edge perception in mammograms: a comparison of two film-screen combinations. AB - The entire skin edge is not always seen on mammograms. The importance of demonstrating the full extent of the breast on a mammogram is uncertain, but there are implications for optimizing film design and use. The new Agfa HDR film has been designed to improve visualization of the skin edge without compromising overall contrast. A quantitative and qualitative comparison between Agfa HDR and the Fuji UMMA film has been performed. A total of 216 mammograms from 54 symptomatic women were analysed. For each view on each woman, one breast was imaged using Fuji UMMA film and the other using Agfa HDR film. The target density achieved on quality control films of PMMA blocks was 1.70 +/- 0.10 for both film types. A radiologist graded the skin edges of all the films as visible, just visible or dark. Measurements were made of the optical densities in the skin edge region and on the fully exposed black part of the film, adjacent to the skin line. 70% of the HDR films had visible skin edges compared with 43% of the UMMA films. 2% of the HDR films had dark skin edges compared with 26% of the UMMA films. Optical densities at the skin edge were broadly similar for the two films and in the range 3.6 +/- 0.3. However, the density of the black part of the Agfa film was about 0.5 higher than with the Fuji film and it appears that this was the main reason the skin edge region was better visualized with the Agfa film. PMID- 10844863 TI - Quality assurance for MRI: practical experience. AB - The aim of this study is to propose guidelines for quality assurance (QA) in MRI, based on a comprehensive assessment of QA parameters undertaken on a busy clinical MRI scanner over the course of 1 year. QA phantoms supplied by the scanner manufacturer were used together with the Eurospin MRI phantom set. Signal to-noise ratio (SNR) and image uniformity were measured daily from spin echo images acquired using a quadrature send-receive head coil and from a gradient echo sequence using the Helmholtz body coil. The voltage of the transmit radiofrequency (RF) amplifier was noted. Monthly measurements of slice thickness, geometric distortion, slice position, image resolution and image ghosting were acquired using the head coil. In addition, SNR was measured monthly on a selection of commonly used coils. Apart from some drift of the RF amplifier voltage, all measurements were within acceptable limits and were stable over the course of 1 year. Satisfactory measurements of SNR were possible using the simple phantom supplied with the scanner. The SNR, geometric distortion and RF amplifier voltage are simple to determine and can be measured in less than 15 min by the scanner operator, using the scanner software. Weekly recording of these parameters is recommended for busy clinical MRI scanners, as this should allow deviations from acceptable limits to be identified early. Such in-house checks can usefully be compared with the less frequent estimations performed by the service engineer. Comprehensive QA routines are discussed for systems used for quantitative measurements. PMID- 10844864 TI - Quality criteria implementation for brain and lumbar spine CT examinations. AB - A study was undertaken to implement the quality criteria proposed by the European Commission for brain general and lumbar spine (disc herniation) CT examinations. The proposed criteria were evaluated for samples including 93 brain and 86 lumbar spine CT examinations, with special emphasis on the diagnostic and radiation dose requirements. The extent to which the image criteria had been achieved was evaluated after two independent observers had each read the images twice. Dose measurements were conducted in parallel to estimate the proposed dose quantities needed to obtain the images. For brain examinations, we found that a group of image criteria were largely met, and met uniformly in all sites. One criterion (1.2.5) was frequently fulfilled but had intermediate values for two sites; the remaining criteria were fulfilled to different extents, although for criteria 1.2.1 and 1.2.2, scores were lower than 50% and 70%, respectively. The mean percentage image quality score had values between 57% and 78%, with variation coefficients in the range 30-68%. Mean values of the dose quantities were in the ranges 44-74 mGy for weighted CT dose index (CTDIw), 497-1018 mGy cm for dose length product (DLP) and 1.1-2.2 mSv for effective dose (E). CTDIw and DLP were not correlated because of significant variations in the scanned length, whereas DLP and E were strongly correlated. A weak relationship between image quality score and DLP was found for the sample as a whole. For lumbar spine examinations, none of the critical reproduction image criteria was systematically achieved. One group of criteria (1.2.7, 1.2.8 and 1.2.9) was fulfilled to a large extent in many departments, but fulfilment of the remainder varied widely. The mean score fluctuated in the range 39-88%, with three groups of differences: low (39-51%), intermediate (67-71%) and high (85-88%). Mean values of the CTDIw varied between sites in the range 27-48 mGy. Mean DLP values varied between 188 mGy cm and 333 mGy cm, and the mean effective dose ranged between 3 mSv and 5 mSv. There were significant differences in effective dose between men and women. By sites, there was no relationship between DLP and mean score, with the highest image score associated with intermediate dose values. The percentage disagreement among the observers about a given criterion ranged between 2% and 22% for brain, and between 3% and 46% for lumbar spine. PMID- 10844865 TI - Reference dose levels for patients undergoing common diagnostic X-ray examinations in Irish hospitals. AB - Wide variations in patient dose for the same type of X-ray examination have been evident from various international dose surveys. Reference dose levels provide a framework to reduce this variability and aid in the optimization of radiation protection. The aim of this study was to establish, for the first time, a baseline for national reference dose levels in Ireland for four of the most common X-ray examinations: chest, abdomen, pelvis and lumbar spine. Measurements of entrance surface dose using thermoluminescent dosemeters (TLDs) for these four X-ray examinations were performed on 10 patients in each of 16 randomly selected hospitals. This represented 42% of Irish hospitals applicable to this study. Results have shown wide variation of mean hospital doses, from a factor of 3 for an anteroposterior lumbar spine to a factor of 23 for the chest X-ray. The difference between maximum and minimum individual patient dose values varied up to a factor of 75. Reasons for these dose variations were complex but, in general, low tube potential, high mAs and low filtration were associated with high-dose hospitals. This study also demonstrated lower reference dose levels of up to 40% when compared with those established by the UK and the Commission of the European Communities for four out of six projections. Only the chest X-ray exhibited a similar reference level to those established elsewhere. This emphasizes the importance of each country establishing its own reference dose levels that are appropriate to their own radiographic techniques and practices in order to optimize patient protection. PMID- 10844866 TI - Dose calculations for asymmetric fields defined by independent collimators using symmetric field data. AB - Several methods have been developed for the dosimetry of asymmetric radiation fields formed by independently moving collimator jaws. Three of these methods, based on different principles and modified to comply with our set of available data, are utilized for the calculation of asymmetric field dose profiles. All three methods use output factors and per cent depth doses or tissue maximum ratios of symmetric fields. In the first method, calculation of the off-centre ratio (OCR) of the asymmetric field is based on the symmetric field from which the asymmetric is originated, by setting the one jaw in an asymmetrical position. In the second method the OCR of the symmetric field is used for the OCR calculation of the asymmetric field of the same size; whereas the third method does not allow for the asymmetric OCR calculation. The results obtained using data for the 6 MV photon beam of a Philips SL-20 linear accelerator indicate that both the first and second method can accurately reproduce asymmetric field profiles from symmetric field data; the third method does not allow for penumbra reproduction, but it is accurate at the central part of the asymmetric field. The problems encountered in the application of the three methods are reported and their accuracy is compared. PMID- 10844867 TI - A digital density equalization technique to improve visualization of breast periphery in mammography. AB - In mammographic imaging, the film area corresponding to the breast periphery is overexposed, resulting in high optical density and degraded contrast in this region. A digital, model-driven density equalization technique was designed and developed to overcome this overexposure problem, taking into account the non linear characteristic curve of the film-digitizer system. The method is based on several image processing and analysis techniques, such as thresholding, which is used to segment the pixels of the mammogram belonging to the breast region from the background, and wavelet-based fusion, which is used to equalize the pixels of breast periphery selectively while leaving the remaining breast region unaffected. Initial application of the method resulted in density-equalized mammographic images, characterized by improved contrast at the breast periphery. PMID- 10844868 TI - Magnetic resonance spectroscopy in the evaluation of neurotoxicity following cranial irradiation for childhood cancer. AB - In order to evaluate the role of proton MR spectroscopy (1H-MRS) in the diagnosis and assessment of long-term radiation-related neurotoxicity, 14 children who had received cranial irradiation for the treatment of childhood leukaemia (n = 6) or brain tumours (n = 8) underwent 1H-MRS, MRI and neuropsychological assessment. Short-term effects at 2 months following treatment were studied in a further three patients. MRI abnormalities were observed in nine patients. No statistically significant differences between patients and controls (n = 17) were seen in any of the calculated 1H-MRS metabolite ratios, in any of the three patient groups. On multivariate logistic regression analysis there was a correlation between the choline/water ratio and a low IQ. It is concluded that any systematic radiation-induced changes in the 1H MRS metabolites must be below the detection threshold of this study. PMID- 10844869 TI - Direct measurement of air kerma rate in air from CDCS J-type caesium-137 therapy sources using a Farmer ionization chamber. AB - A simple method for directly measuring the reference air kerma rate from J-type 137Cs sources using a Farmer 2571 chamber has been evaluated. The method is useful as an independent means of verifying manufacturers' test data. PMID- 10844870 TI - Boron neutron capture therapy of the rat 9L gliosarcoma: evaluation of the effects of shark cartilage. AB - A number of anti-angiogenic substances are now under evaluation, both experimentally and clinically, as potential agents for the treatment of cancer. It has recently been demonstrated that anti-angiogenic agents can increase the therapeutic potential of photon irradiation in a range of tumour models. In the present communication a preliminary assessment is made of the effects of shark cartilage on the response of the rat 9L gliosarcoma to boron neutron capture therapy (BNCT). Shark cartilage was administered orally as an aqueous suspension at a daily dose of approximately 2000 mg kg-1 body weight. The mean survival time of rats receiving no treatment was 20.7 +/- 0.5 days post intracranial tumour implantation. Administration of shark cartilage alone extended the survival time. Two of the rats treated with shark cartilage were healthy and fully active at the end of the evaluation period (43 days post implantation). At autopsy the brain tumours of these animals were a factor of approximately 4 smaller than controls. In a repeat study with shark cartilage alone the survival time was extended by approximately 30%. After boronophenylalanine-mediated BNCT, with or without shark cartilage, the survival time of rats that eventually became moribund was increased by a factor of approximately 2 relative to controls. In both treatment groups approximately 20% of rats were healthy at 1 year after BNCT. There was no evidence of residual tumour at post-mortem. It was concluded that shark cartilage, when given alone, significantly increased the survival time of tumour bearing rats, presumably owing to an anti-angiogenic effect. However, the survival data suggested that boronophenylalanine-mediated BNCT did not appear to be enhanced by the administration of shark cartilage. PMID- 10844871 TI - Benign metastasizing leiomyoma with intracaval leiomyomatosis. AB - We present an unusual case of benign metastasizing leiomyoma in association with intracaval leiomyomatosis. To our knowledge, this is the first reported case of metastasizing leiomyoma with coexistent intravenous leiomyomatosis (IVL). Magnetic resonance imaging is useful for the diagnosis of pelvic and caval IVL. PMID- 10844872 TI - Dynamic susceptibility contrast enhanced MRI in reversible posterior leukoencephalopathy syndrome associated with haemolytic uraemic syndrome. AB - We describe a case of reversible posterior leukoencephalopathy associated with haemolytic uraemic syndrome. Following remission confirmed on MRI, the patient relapsed several months later. Neuroimaging findings on conventional MRI and FLAIR sequences and dynamic susceptibility contrast enhanced MRI are described. White matter abnormalities may be shown on CT or MRI in this syndrome. However, dynamic susceptibility contrast enhanced MRI showed far more extensive abnormality within the brain. In addition, phase contrast angiographic measurement of flow in the carotid and basilar arteries indicated a significant elevation of cerebral blood flow, suggesting a decrease in global cerebrovascular resistance. These observations support existing theories that the disorder is manifested by autoregulatory disturbance in small cerebral vessels. Our findings suggest that this abnormality is far more extensive than is demonstrated on T2 weighted MR images and that it is associated with global abnormality of cerebrovascular autoregulation. PMID- 10844873 TI - The diabetic foot. AB - Foot complications in diabetics often lead to amputation. Ulceration is the most common complication in the diabetic forefoot and underlies more than 90% of cases of pedal osteomyelitis. The diagnosis of osteomyelitis is, nevertheless, difficult, and imaging is an important part of the work-up. Plain radiographs, although useful for anatomical information, are neither sensitive nor specific. Three-phase bone scintigraphy is sensitive but not specific. Labelled leucocyte scintigraphy and MRI are both useful and are complementary to one another. Labelled leucocyte scintigraphy is valuable for diagnosis as well as follow-up of pedal osteomyelitis. MRI offers exquisite anatomical detail, which is invaluable for guiding surgical management. The principal complication in the mid and hind foot is the neuropathic or Charcot joint. Although infection of the neuropathic joint is infrequent, its diagnosis is difficult. The extensive bony changes that accompany this disorder severely diminish the value of radiography and bone scintigraphy. It is not always possible to distinguish the marrow oedema of neuropathy from that of osteomyelitis and the role of MRI in the evaluation of this entity is still uncertain. Uptake of labelled leucocytes in the absence of infection may occur and is owing, at least in part, to haematopoietically active marrow. Combined leucocyte/marrow scintigraphy holds considerable promise for identifying the infected Charcot joint. PMID- 10844874 TI - Case of the month. Intrapulmonary or extrapulmonary? PMID- 10844875 TI - Comments on the British Institute of Radiology Working Party advice for patients after radionuclide therapy. PMID- 10844876 TI - Paediatric sedation using quinalbarbitone. PMID- 10844877 TI - Of knowledge and deception. PMID- 10844878 TI - A randomized controlled study of reviewer bias against an unconventional therapy. AB - A study was designed to test the hypothesis that experts who review papers for publication are prejudiced against an unconventional form of therapy. Two versions were produced (A and B) of a 'short report' that related to treatments of obesity, identical except for the nature of the intervention. Version A related to an orthodox treatment, version B to an unconventional treatment. 398 reviewers were randomized to receive one or the other version for peer review. The primary outcomes were the reviewers' rating of 'importance' on a scale of 1-5 and their verdict regarding rejection or acceptance of the paper. Reviewers were unaware that they were taking part in a study. The overall response rate was 41.7%, and 141 assessment forms were suitable for statistical evaluation. After dichotomization of the rating scale, a significant difference in favour of the orthodox version with an odds ratio of 3.01 (95% confidence interval, 1.03 to 8.25), was found. This observation mirrored that of the visual analogue scale for which the respective medians and interquartile ranges were 67% (51% to 78.5%) for version A and 57% (29.7% to 72.6%) for version B. Reviewers showed a wide range of responses to both versions of the paper, with a significant bias in favour of the orthodox version. Authors of technically good unconventional papers may therefore be at a disadvantage in the peer review process. Yet the effect is probably too small to preclude publication of their work in peer-reviewed orthodox journals. PMID- 10844879 TI - Clinical questions and information needs. PMID- 10844880 TI - The gripe water story. PMID- 10844881 TI - Clinical management of glaucoma. PMID- 10844882 TI - Lipid abnormalities in renal disease. PMID- 10844883 TI - Who looks after people with diabetes: primary or secondary care? AB - Because the prevalence of type 2 diabetes has increased greatly over the past decade, UK general practitioners have been encouraged to develop services for people with diabetes and to offer structured diabetes care. The resultant shift from secondary care can place considerable demands on primary health care teams. Data were obtained from 108 practices in two English health districts followed up in primary and secondary care. Nearly two-thirds of the people with diabetes were being followed up only in general practice, the remainder in hospital or both. The proportion managed in primary care varied from 5.6% to 94.6%. The settings where diabetes care was most likely to be offered were training practices, practices with good nursing support, practices with a high prevalence of diabetes, and practices in which a high proportion of diabetic patients were controlled by diet or hypoglycaemic agents. Tight control of glycaemia and blood pressure is now seen as important in diabetes, and is best achieved in general practice. This survey revealed large variations in delivery of general-practice diabetes care that need to be addressed by better organization and funding. PMID- 10844884 TI - Domiciliary visits to the old and the mentally ill: how valuable? AB - In the early days of the British National Health Service, domiciliary visits were a continuation of the tradition whereby general practitioners (GPs) met consultants in the patient's home. The nature of domiciliary visits, which still attract a special fee, has since changed. We analysed the effectiveness of all domiciliary visits undertaken in a NHS trust providing primary care, mental health and elderly care services to a population of 470,000. Data were obtained from domiciliary visit claim forms and from questionnaires completed by the consultant, the referring GP and consultant peer reviewers. The largest number of visits (total 234) was in geriatric medicine 48.9%, followed by old-age psychiatry 44.9%. Geriatric medicine was more likely than psychiatry to admit patients to hospital (19%) after a visit. All domiciliary visits in old-age psychiatry were done during the day (9 am to 5 pm). Only 2% of GPs stated that they attended any of the domiciliary visits; almost all thought that the outcome of domiciliary visits was of value. Old-age-psychiatry peer reviewers believed that all visits in that specialty were appropriate; in geriatric medicine this figure was 77% and in other psychiatric specialties 65%. The findings indicate that domiciliary visits were not being used routinely as a pathway to hospital admission, though they were often used to expedite admission or gain a quick consultant opinion; the visits were valued by GPs. The practice of domiciliary visits differs greatly from the definition in NHS terms and conditions of service. One or other should be altered. PMID- 10844885 TI - Ophthalmic zoster sine herpete. PMID- 10844887 TI - Hereditary Madelung's disease. PMID- 10844886 TI - Appendiceal adenocarcinoma with ovarian metastases in the third trimester of pregnancy. PMID- 10844888 TI - Abdominal actinomycosis presenting as psoas abscess. PMID- 10844889 TI - Isolated sarcoidosis of the breast. PMID- 10844890 TI - The emperor with the shaking head: Claudius' movement disorder. PMID- 10844891 TI - Birth defects described in Elizabethan ballads. PMID- 10844892 TI - The most pressing issue. PMID- 10844893 TI - The most pressing issue. PMID- 10844894 TI - Pulsating veins. PMID- 10844895 TI - Biblical origins of placebo. PMID- 10844896 TI - Red jackets and red noses. PMID- 10844897 TI - Accountability, clinical governance and the acceptance of imperfection. PMID- 10844898 TI - Accountability, clinical governance and the acceptance of imperfection. PMID- 10844899 TI - A national database of medical error. PMID- 10844900 TI - A national database of medical error. PMID- 10844901 TI - Postmortem and perimortem caesarean section. PMID- 10844902 TI - Intracranial hypertension. PMID- 10844903 TI - Herpes simplex type 2 infections--an update. PMID- 10844904 TI - The blind leading the blind. PMID- 10844905 TI - ST elevation in a man with a gunshot wound to the head. PMID- 10844906 TI - A student with renal failure and cardiac consequences. PMID- 10844907 TI - Selecting treatment in patients with epilepsy. PMID- 10844908 TI - Adjuvant therapy of colorectal cancer. PMID- 10844909 TI - Food allergy: from biology toward therapy. PMID- 10844910 TI - Spontaneous bacterial peritonitis: keys to management. PMID- 10844911 TI - Genetics in practice: 1. Hereditary hemochromatosis. PMID- 10844912 TI - The bone and joint decade: 2000-2010. PMID- 10844913 TI - Evaluation of genetic testing for hereditary non-polyposis colorectal cancer (HNPCC) PMID- 10844914 TI - Are patients with lung cancer the poor relations in oncology? PMID- 10844915 TI - Routine antenatal screening: a need to evaluate Australian practice. PMID- 10844916 TI - Value of predictive genetic testing in management of hereditary non-polyposis colorectal cancer (HNPCC) AB - OBJECTIVE: To investigate the impact of predictive genetic testing on colonoscopic surveillance in an extended family with hereditary non-polyposis colorectal cancer (HNPCC). SETTING: Familial Bowel Cancer Service, The Royal Melbourne Hospital, Victoria. SUBJECTS: 96 people registered with the Service who were apparently unaffected members of an extended family that met the classic Amsterdam criteria for HNPCC and carried an MLH1 gene mutation (IVS9 + 3insT). INTERVENTION: Predictive genetic testing was offered in a cascade manner to at risk family members; mutation-positive individuals were advised to have annual colonoscopic surveillance, while mutation-negative individuals were withdrawn from surveillance. MAIN OUTCOME MEASURES: Previous compliance with recommended colonoscopic surveillance; uptake and results of genetic testing; expected effect of genetic test results on number of colonoscopies over five years. RESULTS: 22 of the 96 family members (23%) were not complying with recommended surveillance. Of 48 individuals offered predictive genetic testing, 41 (85%) responded and 39 (81%) underwent testing. Seven of the 39 (18%) were positive for the family specific mutation, and 32 (82%) were negative. The 39 tested individuals and 37 of their descendants who were registered with the screening program had undergone 70 colonoscopies in the five years before genetic testing. In the five years after testing, only 37 surveillance colonoscopies were planned (annual or two yearly colonoscopies for the six mutation-positive individuals and five-yearly colonoscopies for four mutation-negative individuals with previously identified adenoma), an almost 50% reduction in colonoscopies. CONCLUSION: Predictive genetic testing in HNPCC families allows many individuals to be withdrawn from regular colonoscopic surveillance. It may therefore reduce costs, as well as have emotional benefits for many individuals. PMID- 10844917 TI - Prevention of perinatal group B streptococcal disease: screening practice in public hospitals in Victoria. AB - OBJECTIVES: To survey clinical protocols for prevention of early-onset group B streptococcal disease (EOGBSD) of the newborn in public maternity hospitals. DESIGN: Postal questionnaire with telephone follow-up when required. SETTING: All hospitals that undertook deliveries in public patients in the State of Victoria, November 1997 to January 1998. RESULTS: The survey was sent to 84 hospitals: 71 responded and 64 met the criteria and provided usable data (76% response rate). These 64 represented 42,784 births (68% of births in Victoria in 1996). Most hospitals (62; 97%) undertook actions that would identify and treat pregnant women at risk of EOGBSD. 48 (75%) performed bacteriological screening for maternal GBS carriage, but only 20 of these had a unified protocol. Screening was mostly by low vaginal swab (15 hospitals) and before 30 weeks' gestation (12 hospitals). Low vaginal swab plus anal swab was used in only one hospital. Bacteriological screening was significantly more common in metropolitan hospitals than in rural hospitals (100% versus 67%; P = 0.007, Fisher's exact test). Targeting of prophylaxis by recognised risk factors was reported by 59 (92%) hospitals, 45 of which also undertook screening. There was considerable variation in the specific risk factors used. CONCLUSIONS: While there was clearly widespread awareness of EOGBSD in Victorian public hospitals, prevention programs varied considerably. The development of consensus practice guidelines might improve EOGBSD prevention, reducing morbidity, mortality and costs. PMID- 10844918 TI - Reported management of lung cancer in Victoria in 1993: comparison with best practice. Anti-Cancer Council of Victoria Lung Cancer Study Group. AB - OBJECTIVE: To determine recent patterns of management of lung cancer in Victoria in order to stimulate interest in the development of Australian consensus guidelines. DESIGN: A cross-sectional survey of doctors responsible for the care of an incident series of lung cancer patients in 1996-1997. PARTICIPANTS: 1054 people diagnosed with primary lung cancer in the State of Victoria between 1 January 1993 and 31 July 1993 and notified to the Victorian Cancer Registry. MAIN OUTCOME MEASURES: Method of diagnosis; tumour characteristics; factors affecting management plan; first-line and subsequent treatment; outcome; and patients' current status. RESULTS: Questionnaires were completed for 868 eligible patients (82%): 635 (73%) diagnosed with non-small-cell lung cancer, 124 (14%) diagnosed with small-cell lung cancer, and 109 (13%) with no histological diagnosis. Chest x-ray (814 patients; 94%) and computed tomography (CT) of the chest and abdomen (589 patients; 68%) were the most common investigations, and was the only diagnostic procedure in 48 patients (6%). Treatments were radiotherapy alone or in combination (385 patients; 44%), surgery alone or in combination (196 patients; 23%), chemotherapy alone or in combination (152 patients; 18%); 215 patients (25%) received no antitumour therapy. 243 patients (28%) were treated initially with curative intent. A further 399 (46%) were treated initially with palliative intent, and in 219 (55%) of these good symptom control was achieved. For 427 patients (49%) tumour size was not recorded. While 23% of non-small-cell patients had limited disease, only 8% were investigated with mediastinoscopy. Only four patients (13%) with limited-stage, small-cell lung cancer had combined modality treatment. There was little use of adjuvant chemotherapy or neoadjuvant therapy. The five-year crude survival rate was 11%. CONCLUSIONS: The demographics of lung cancer in Victoria are similar to other population-based studies. Patterns of management are not uniform, and are inconsistent with current published guidelines. PMID- 10844919 TI - Notification of infectious diseases by general practitioners: a quantitative and qualitative study. AB - OBJECTIVE: To examine general practitioners' knowledge and practices concerning reporting of notifiable conditions, and to examine barriers to notification of infectious diseases by general practitioners and identify strategies for improving the notification process. DESIGN AND SETTING: Audit of the 100 most recent notifications received by the South Eastern Sydney Public Health Unit of cases of each of hepatitis A, pertussis and measles; and focus groups with GPs practising in Sydney's eastern and southern suburbs, some of whom were selected on the basis of their notification practices. RESULTS: Although these diseases are notifiable on clinical suspicion, only about 40% of the hepatitis A and pertussis cases and 80% of measles cases (54% overall) had been notified by GPs. Delays between doctor and laboratory notifications were an average of seven days for hepatitis A, 19 days for pertussis and seven days for measles. Focus groups showed that at least some GPs have poor understanding of the process of notification, most felt uncomfortable notifying an unconfirmed case, many preferred to leave notification to the laboratory because of concerns about damaging the doctor-patient relationship, and that there is need for financial or other incentives. CONCLUSIONS: There are deficiencies in the completeness and timeliness of notification by GPs which may adversely affect the timing of prophylaxis and outbreak control. Notification by GPs may be improved by such strategies as better notification forms and better feedback to doctors on the outcomes. PMID- 10844920 TI - Healthcare rationing in The Netherlands: the need for specific guidelines. PMID- 10844921 TI - Medicine for the millennium: the challenge of postmodernism. AB - As the new millennium dawns, Australian society is becoming more post-modern, whereas the medical system remains increasingly modernist in its outlook. In this article, we discuss the emerging prevalence of post-modernism and examine current medical education and practice strategies, such as evidence-based medicine, from a post-modern perspective. We argue that if medicine does not respond to the ideas of post-modernism, which challenges the concepts of truth and our ability to be objective, it may become increasingly irrelevant to the needs of a changing society. PMID- 10844922 TI - The hospitalist: a third alternative. PMID- 10844923 TI - Genetic testing for Alzheimer's disease. AB - Genetic factors are important in the development of Alzheimer's disease (AD). Familial AD can result from rare mutations in some genes. Other genes, such as the apolipoprotein E gene (APOE), operate as risk factors for late-onset sporadic AD. On a background of advances in the genetics of AD we suggest a way in which genetic information may be used in the diagnosis of AD. If there is a positive family history of early-onset dementia and the clinical features suggest AD, patients may be tested for presenilin and amyloid precursor protein gene mutations with appropriate pretest and post-test counselling. Predictive testing should be performed under guidelines developed by the World Federation of Neurology and the Human Genetics Society of Australasia. The usefulness of APOE genotyping as an adjunct to conventional diagnostic tests is unknown; data suggest it has low sensitivity and specificity and may have little predictive value in an individual patient. APOE genotyping should not be performed in asymptomatic individuals, except as part of an ethically approved research project; this recommendation is supported by a number of international consensus statements. APOE testing should not be used as a diagnostic test without adequate pretest and post-test counselling, education and support. APOE testing should not be used as a sole diagnostic test in the work-up of patients with AD. Genetic risk factors other than APOE require validation and should not be used routinely, except as part of an ethically approved research protocol. PMID- 10844924 TI - The role of oestrogen in cardiovascular disease: benefit or harm? PMID- 10844925 TI - Iodine status of Australia: look before we leap! PMID- 10844926 TI - Late diagnosis of three cases of transfusion-acquired HIV-1 infection. PMID- 10844927 TI - Patterns of drowning in Australia, 1992-1997. PMID- 10844928 TI - The Clinical Information Access Project. PMID- 10844929 TI - Integrated electronic health records. PMID- 10844930 TI - Internet use by radiation oncology patients: a pilot study. PMID- 10844931 TI - Lack of evidence of risk from low-level radiation. PMID- 10844932 TI - Risk of radiation-induced cancer at low doses and low dose rates. PMID- 10844933 TI - Advanced distal colonic lesions as predictors of advanced lesions in the proximal colon. AB - The characteristics of adenomas found during sigmoidoscopy have been suggested to predict synchronous adenomas in the proximal colon. Our aim was to examine whether the presence and characteristics of distal colonic lesions are associated with the presence and characteristics of lesions in the proximal colon. We performed a university hospital based case-control study with 3,268 consecutive subjects who received colonoscopy between January 1992 and December 1995. Subjects who had a history of colonic polyps, inflammatory bowel disease, intestinal resection, or had a contraindication against biopsies were excluded. Number size, and histologic characteristics of polyps in the distal and proximal colon were recorded. Advanced lesions were defined as neoplastic lesions with 1 or more of the following features: 1) > or = 1 cm diameter, and/or 2) villous histology, and/or 3) severe dysplasia or carcinoma, and/or 4) > or = 3 lesions. We found that 439 patients had neoplastic lesions in the distal colon only (61.3% with advanced lesions), 198 patients had lesions in the proximal colon only (55.1% advanced), and 197 had lesions in both the distal colon (74.6% advanced) and the proximal colon (55.8% advanced). Distal lesions were significantly more often advanced in patients with synchronous proximal lesions compared with patients with lesions in the distal colon only (odds ratio: 1.9; 95% confidence interval [CI]: 1.3-2.8; p < 0.001). The odds ratios for finding any neoplastic lesion in the proximal colon and an advanced proximal lesion, respectively, were 3.7 (2.6-5.3) (p < 0.001) and 2.2 (1.3-3.7) (p < 0.01) when a nonadvanced lesion was found in the distal colon, and 6.8 (5.3-8.7) (p < 0.001) and 6.7 (4.9-9.0) (p < 0.001) when an advanced lesion was found in the distal colon. Logistic regression analysis revealed number of distal polyps and villous histology as independent predictors of advanced lesions in the proximal colon; size and severe dysplasia were not independent predictors. In conclusion, characteristics of neoplastic lesions in the distal colon predict the presence and characteristics of lesions in the proximal colon. PMID- 10844934 TI - Adult Fanconi syndrome secondary to light chain gammopathy. Clinicopathologic heterogeneity and unusual features in 11 patients. AB - Fifty-seven cases of Ig light chain-associated Fanconi syndrome (FS) have been reported so far, mostly as isolated reports. The pioneering work by Maldonado and associates (35), who reviewed the first 17 cases in 1975, led to the unifying concept that patients with FS and Bence Jones proteinuria have a special form of plasma cell dyscrasia characterized by slow progression of the tumor and by prominent crystal formation in proximal tubule cells, in the absence of myeloma casts in the distal tubule. We carefully reappraised these characteristics in a series of 11 patients. Ten renal biopsy specimens were available for electron microscopy, adding to the 15 previously reported cases with ultrastructural studies. Moreover, 10 of the kappa light chains could be entirely or partially sequenced and tested for their resistance to cathepsin B, a lysosomal protease present in proximal tubule cells. Our series showed an unexpected clinicopathologic heterogeneity. Seven patients presented with the typical clinical and pathologic features of FS and low-mass myeloma or monoclonal gammopathy of undetermined significance (MGUS), in keeping with Maldonado et al's description. Crystals in bone marrow cells were detected in patients of this group, only. Three patients who presented with full-blown FS exhibited, however, the characteristic features of myeloma cast nephropathy in the setting of high mass myeloma. One patient of this group also had numerous crystals in proximal tubule cells. The eleventh patient had complete FS with MGUS, but no crystals in proximal tubule cells even after electron microscopy. Contrasting with the clinicopathologic heterogeneity, genetic and biochemical analyses of the light chains showed a striking homogeneity. First, they all were of the kappa type. Second, 8 of 9 belonged to the V kappa I variability subgroup, which indicates that FS light chains are related by the sequence of their variable regions. Third, the 8 V kappa I light chain sequences most likely originated from only 2 germline genes, LCO2/012 and LCO8/018. Fourth, all 5 LCO2/012-derived sequences presented an unusual hydrophobic or nonpolar residue at position 30. These sequence peculiarities may account for unusual physicochemical properties of the light chains including the resistance of their variable domain V kappa to proteolysis by cathepsin B, observed in 7 of 9 patients in our series, while light chains isolated from patients with myeloma cast nephropathy are completely digested. Resistance of V kappa to proteolysis in FS patients can explain the accumulation of the light chain in the endocytotic compartment of the proximal tubule cells, leading to impairment of proximal tubule functions. PMID- 10844935 TI - Chronic granulomatous disease. Report on a national registry of 368 patients. AB - A registry of United States residents with chronic granulomatous disease (CGD) was established in 1993 in order to estimate the minimum incidence of this uncommon primary immunodeficiency disease and characterize its epidemiologic and clinical features. To date, 368 patients have been registered; 259 have the X linked recessive form of CGD, 81 have 1 of the autosomal recessive forms, and in 28 the mode of inheritance is unknown. The minimum estimate of birth rate is between 1/200,000 and 1/250,000 live births for the period 1980-1989. Pneumonia was the most prevalent infection (79% of patients; Aspergillus most prevalent cause), followed by suppurative adenitis (53% of patients; Staphylococcus most prevalent cause), subcutaneous abscess (42% of patients; Staphylococcus most prevalent cause), liver abscess (27% of patients; Staphylococcus most prevalent cause), osteomyelitis (25% of patients; Serratia most prevalent cause), and sepsis (18% of patients; Salmonella most prevalent cause). Fifteen percent of patients had gastric outlet obstruction, 10% urinary tract obstruction, and 17% colitis/enteritis. Ten percent of X-linked recessive kindreds and 3% of autosomal recessive kindreds had family members with lupus. Eighteen percent of patients either were deceased when registered or died after being registered. The most common causes of death were pneumonia and/or sepsis due to Aspergillus (23 patients) or Burkholderia cepacia (12 patients). Patients with the X-linked recessive form of the disease appear to have a more serious clinical phenotype than patients with the autosomal recessive forms of the disease, based on the fact that they are diagnosed significantly earlier (mean, 3.01 years of age versus 7.81 years of age, respectively), have a significantly higher prevalence of perirectal abscess (17% versus 7%), suppurative adenitis (59% versus 32%), bacteremia/fungemia (21% versus 10%), gastric obstruction (19% versus 5%), and urinary tract obstruction (11% versus 3%), and a higher mortality (21.2% versus 8.6%). PMID- 10844937 TI - Doing the right thing for US healthcare. PMID- 10844936 TI - Genetic, biochemical, and clinical features of chronic granulomatous disease. AB - The reduced nicotinamide dinucleotide phosphate (NADPH) oxidase complex allows phagocytes to rapidly convert O2 to superoxide anion which then generates other antimicrobial reactive oxygen intermediates, such as H2O2, hydroxyl anion, and peroxynitrite anion. Chronic granulomatous disease (CGD) results from a defect in any of the 4 subunits of the NADPH oxidase and is characterized by recurrent life threatening bacterial and fungal infections and abnormal tissue granuloma formation. Activation of the NADPH oxidase requires translocation of the cytosolic subunits p47phox (phagocyte oxidase), p67phox, and the low molecular weight GT-Pase Rac, to the membrane-bound flavocytochrome, a heterodimer composed of the heavy chain gp91phox and the light chain p22phox. This complex transfers electrons from NADPH on the cytoplasmic side to O2 on the vacuolar or extracellular side, thereby generating superoxide anion. Activation of the NADPH oxidase requires complex rearrangements between the protein subunits, which are in part mediated by noncovalent binding between src-homology 3 domains (SH3 domains) and proline-rich motifs. Outpatient management of CGD patients relies on the use of prophylactic antibiotics and interferon-gamma. When infection is suspected, aggressive effort to obtain culture material is required. Treatment of infections involves prolonged use of systemic antibiotics, surgical debridement when feasible, and, in severe infections, use of granulocyte transfusions. Mouse knockout models of CGD have been created in which to examine aspects of pathophysiology and therapy. Gene therapy and bone marrow transplantation trials in CGD patients are ongoing and show great promise. PMID- 10844938 TI - Periodontal infection a risk factor for cardiovascular disease? PMID- 10844939 TI - Protecting your system from digital demons. Software vaccines to help you fight off viruses. PMID- 10844940 TI - Evidence-based therapy for type 2 diabetes. The best and worst of times. AB - Type 2 diabetes, which is increasing in prevalence, is a multifaceted disease that presents a challenge to the primary care physician. In this article, Drs Flemmer and Vinik review the significant findings of several important clinical trials and present a method of comprehensive staged management to improve the quality of patient care. PMID- 10844941 TI - Large, patchy skin eruptions after a hiking trip. Erythema chronicum migrans hallmarks Lyme disease. PMID- 10844942 TI - Contributing factors in resistant hypertension. Truly refractory disease is rarely found in a properly conducted workup. AB - Systematic evaluation of each patient with refractory hypertension is necessary to identify conditions or altered mechanisms of hypertension that are amenable to targeted therapy. Issues that should be carefully investigated are appropriateness of the regimen, possible drug interactions, patient compliance, associated conditions (alcohol or recreational drug use, hyperinsulinemia), pseudohypertension, office hypertension, and volume overload. When these problems are eliminated, causes of secondary hypertension should be sought, the most common being coarctation of the aorta, Cushing's syndrome, primary aldosteronism, pheochromocytoma, renovascular disease, thyroid and parathyroid disease, and renal parenchymal disease. In a few cases, a careful hemodynamic and neurohumoral assessment is needed to direct treatment. When the recommended stepwise workup is followed, hypertension that is truly refractory to treatment is a rare finding. PMID- 10844943 TI - Hyponatremia and hypernatremia. A systematic approach to causes and their correction. AB - Disorders of sodium and water metabolism can be approached by following a few basic steps: Thorough history taking and physical examination that focuses on volume assessment and laboratory evaluation that includes serum osmolality, urine osmolality, and urine sodium concentration are usually all that are required for diagnosis. Results of these findings are helpful in guiding therapy. Monitoring serum sodium concentration often to ensure adequate treatment and to avoid potential complications is required in management of both hyponatremia and hypernatremia. PMID- 10844944 TI - Ominous sign of a disfiguring disease. Leprosy (Hansen's disease). PMID- 10844945 TI - Colorectal cancer 2000. Education and screening are essential if outcomes are to improve. AB - Mortality from colorectal cancer is less threatening than it was 10 years ago, but this cancer is still a leading cause of death in the United States and the world. Careful history taking, education about the importance of a high-fiber, low-fat diet and exercise, and regular screening are important steps in improving outcomes. For the busy primary care physician and the equally busy patient, prevention can become a low priority. However, the benefits of early preventive intervention are clearly worth the effort. PMID- 10844946 TI - Today's approach to esophageal cancer. What is the role of the primary care physician? AB - Esophageal cancer is a devastating disease. Patients are most likely to first present to a primary care physician, who should immediately undertake an initial diagnostic and staging evaluation. The approach then should become multidisciplinary, involving surgical and medical oncologists, radiation oncologists, and gastroenterologists. Optimal therapy of esophageal cancer continues to evolve. While multimodality therapy remains controversial, studies are ongoing to determine the best approaches to improve survival. Further studies to define the role of surveillance endoscopy in certain high-risk patients are warranted. PMID- 10844947 TI - Hepatocellular carcinoma. A concise guide to its status and management. AB - Although common worldwide, hepatocellular carcinoma is relatively rare in the United States. However, for unknown reasons, the incidence is rising. Multiple causes exist, but chronic viral hepatitis in the setting of cirrhosis is probably the most common. Despite limitations, AFP measurement and multiple-phase abdominal CT are the most sensitive tests for diagnosis. Surgical resection and liver transplantation are at present the only treatment options that offer potential for long-term survival or cure in limited-stage hepatocellular carcinoma. Otherwise, the prognosis is poor, and 1-year survival is rare. Future efforts should focus on improving detection of early-stage disease and improving preventive measures to reduce viral hepatitis infection, transmission, and progression. PMID- 10844948 TI - Is it premenstrual syndrome? Keys to focused diagnosis, therapies for multiple symptoms. AB - Premenstrual syndrome (PMS) is sometimes facetiously blamed when a woman behaves erratically, when in fact, the cyclic symptoms of true PMS do cause significant difficulties in everyday life and relationships. In this article, Drs Frye and Silverman outline the criteria for diagnosing PMS and premenstrual dysphoric disorder and describe medications, nutritional supplements, and behavior modifications that have proved effective for relief of PMS symptoms. PMID- 10844949 TI - Colorectal cancer. PMID- 10844950 TI - Veterinarians: quiet contributors to nation's policy. PMID- 10844951 TI - AAHA says national commission must be 'catalyst for change'. PMID- 10844952 TI - Pet health insurance: what veterinarians should know. PMID- 10844953 TI - Paratuberculosis regulations changed. PMID- 10844954 TI - AVMA comments on permethrin toxicosis. PMID- 10844955 TI - AVMA backs New Markets for State Inspected Meat Act of 1999. PMID- 10844956 TI - USDA authorizes funding for scrapie eradication. PMID- 10844957 TI - Ethical concerns about communicators. PMID- 10844958 TI - Looking for leaders. PMID- 10844959 TI - Treatment options needed. PMID- 10844960 TI - Recombinant bovine somatotropin and animal welfare. PMID- 10844961 TI - What is your diagnosis? Circumferential foreign body within the soft t tissues of the neck and pronounced tracheal compression. PMID- 10844962 TI - Is there an oversupply of veterinarians? PMID- 10844964 TI - Animal theft. PMID- 10844963 TI - Dietary considerations for dogs and cats with renal disease. PMID- 10844965 TI - Veterinary photodynamic therapy. PMID- 10844966 TI - Current and future trends in demographics of veterinary medicine in California. AB - OBJECTIVE: To evaluate the present and future supply of veterinarians in California, in light of changing trends in animal ownership. DESIGN: Database analysis. SAMPLE POPULATION: Human and animal populations, including populations of veterinarians, throughout the United States. PROCEDURES: Data on animal and human populations were compiled from a number of sources, including the US Census Bureau, American Veterinary Medical Association, State of California Department of Finance, and State of California Veterinary Medical Board. The distribution of veterinarians in California was contrasted with other health professionals in California and with that of veterinarians in other states. Recent changes in veterinary medical demographics in California were quantified and used to develop in-state projections about the supply of veterinarians for the next 20 years. RESULTS: Although California is the most populous of the 50 states, only 7 states had fewer veterinarians per capita. Furthermore, California ranked next to last among states in increase of number of veterinarians between 1990 and 1995. Los Angeles County had the smallest per-capita number of veterinarians among 9 populous California counties. During that period, California had a net gain of only 6 veterinarians who were exclusively or predominantly large-animal or mixed animal practitioners. CONCLUSIONS AND CLINICAL RELEVANCE: If current trends continue, the per-capita number of veterinarians will continue to decrease in California. To maintain the current ratio of 17.8 veterinarians/100,000 people in California in the future, we estimate that an additional 50 veterinarians above the currently predicted increase will be required annually. PMID- 10844967 TI - Analysis of cerebrospinal fluid from dogs and cats after 24 and 48 hours of storage. AB - OBJECTIVE: To compare differential cell counts and cell characteristics of CSF samples analyzed immediately or after storage for 24 and 48 hours at 4 C with and without the addition of autologous serum. DESIGN: Prospective study. ANIMALS: 36 dogs and 6 cats. PROCEDURE: CSF samples were collected from the cerebellomedullary cistern and divided into 250-microliter aliquots. Slides of CSF samples were prepared by use of cytocentrifugation immediately and after 24 and 48 hours of storage with addition of autologous serum (final concentrations, 11 and 29%). Differential cell counts and number of unrecognizable cells were compared among preparations. RESULTS: Significant differences in the differential cell counts were not detected among samples analyzed before or after storage. Although the number of unrecognizable cells increased with storage time, this did not result in a significant effect on cell distribution or diagnosis. Cells in CSF samples stored with 11% serum more closely resembled cells in fresh samples than did cells in samples stored with 29% serum. CONCLUSIONS AND CLINICAL RELEVANCE: CSF samples collected at veterinary clinics remote from a diagnostic laboratory or during nonoperational hours may be preserved through the addition of autologous serum. Evaluation of such samples is likely to result in an accurate diagnosis for at least 48 hours after collection. PMID- 10844968 TI - Prevalence of pituitary tumors among diabetic cats with insulin resistance. AB - OBJECTIVE: To determine prevalence of pituitary tumors, detectable by means of computed tomography or magnetic resonance imaging, in cats with insulin resistance suspected to have acromegaly or hyperadrenocorticism versus cats with well-controlled diabetes mellitus. DESIGN: Case series. ANIMALS: 16 cats with insulin resistance that were also suspected to have acromegaly (n = 12) or pituitary-dependent hyperadrenocorticism (4) and 8 cats with well-controlled diabetes mellitus. PROCEDURE: Computed tomography was performed on all 16 cats with insulin resistance and 2 cats in which diabetes mellitus was well controlled. The remaining 6 cats in which diabetes mellitus was well-controlled underwent magnetic resonance imaging. Images were obtained before and immediately after i.v. administration of contrast medium. RESULTS: Computed tomography revealed a mass in the region of the pituitary gland in all 16 cats with insulin resistance. Maximum width of the masses ranged from 4.4 to 12.7 mm; maximum height ranged from 3.1 to 12.6 mm. Results of computed tomography performed on 2 cats with well-controlled diabetes and magnetic resonance imaging performed on the remaining 6 cats were considered normal. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that cats with insulin resistance suspected to have acromegaly or pituitary-dependent hyperadrenocorticism are likely to have a pituitary mass detectable by means of computed tomography or magnetic resonance imaging. PMID- 10844969 TI - Association between postoperative outcome and results of magnetic resonance imaging and computed tomography in working dogs with degenerative lumbosacral stenosis. AB - OBJECTIVE: To determine whether results of magnetic resonance imaging (MRI) and computed tomography (CT) are associated with postoperative outcome in working dogs with degenerative lumbosacral stenosis. DESIGN: Prospective cohort study. ANIMALS: 12 dogs treated surgically for degenerative lumbosacral stenosis. PROCEDURE: The lumbosacral vertebral column was examined before surgery by use of MRI and CT and after surgery by use of CT. Outcome, based on performance in standardized training exercises, was assessed 6 months after decompressive surgery. Associations between imaging results and postoperative outcome were determined by use of a Fisher exact test and logistic regression. RESULTS: None of the dogs were able to perform their duties before surgery. By 6 months after surgery, 8 of 12 dogs had been returned to full active duty. Nerve tissue compression was effectively localized by use of CT and MRI. Significant associations between results of imaging studies and postoperative outcome were not identified. CONCLUSIONS AND CLINICAL RELEVANCE: Surgical intervention is justified in high-performance working dogs with degenerative lumbosacral stenosis. However, results of imaging studies may be less important than clinical or surgical factors for predicting outcome in affected dogs. PMID- 10844970 TI - Prognostic factors and patterns of treatment failure in dogs with unresectable differentiated thyroid carcinomas treated with megavoltage irradiation. AB - OBJECTIVE: To determine quality and duration of progression-free survival (PFS) time in dogs with unresectable thyroid carcinomas treated with definitive megavoltage irradiation and analyze prognostic factors of PFS and patterns of failure (local recurrence vs metastasis). DESIGN: Prospective clinical trial. ANIMALS: 25 dogs with locally advanced thyroid carcinomas and no evidence of metastasis. PROCEDURE: Dogs were treated with 48 Gy during 4 weeks on an alternate-day schedule of 4 Gy/fraction. RESULTS: Irradiation was safe and effective for treatment of large unresectable thyroid carcinomas. Progression free survival rates were 80% at 1 year and 72% at 3 years. Time to maximum tumor size reduction ranged from 8 to 22 months. Factors affecting PFS were not found. Twenty-eight percent (7/25) of dogs developed metastasis. Dogs with bilateral tumors had 16 times the risk of developing metastases, compared with dogs with a single tumor. Dogs with no evidence of tumor progression had 15 times less risk of developing metastases. Radiation-induced hypothyroidism was suspected in 2 dogs 13 and 29 months after irradiation. CONCLUSIONS AND CLINICAL RELEVANCE: Irradiation is effective for local control of thyroid tumors, despite their slow regression rate. Results provided evidence that local tumor control affects metastatic outcome in dogs with thyroid carcinomas and is a strong basis for the development of new approaches that include irradiation in the management of dogs with advanced thyroid carcinomas. Improvements in local tumor control alone may be insufficient to improve survival times because of the high risk of metastatic spread before an initial diagnosis is made, which warrants initiation of early systemic treatment. PMID- 10844971 TI - Development of glaucoma after cataract surgery in dogs: 220 cases (1987-1998). AB - OBJECTIVE: To examine postoperative ocular hypertension (POH) and other variables as predictors of the risk of developing glaucoma after cataract surgery in dogs. DESIGN: Retrospective study. ANIMALS: 220 dogs that had cataract surgery. PROCEDURE: Medical records of 220 dogs (346 eyes) that had extracapsular cataract removal or phacoemulsification of cataracts were reviewed. With respect to glaucoma development, 8 variables were analyzed, which included development of POH, breed, sex, age at time of surgery, eye (right vs left), phacoemulsification time, intraocular lens (IOL) placement (yes or no), and stage of cataract development. Eyes developed glaucoma within 6 or 12 months of surgery or did not have signs of glaucoma at least 6 or 12 months after cataract surgery. RESULTS: Of 346 eyes, 58 (16.8%) developed glaucoma after surgery. At 6 months, 32 of 206 (15.5%) eyes examined had glaucoma; at 12 months, 44 of 153 (28.8%) eyes examined had glaucoma. Median follow-up time was 5.8 months (range, 0.1 to 48 months). Mixed-breed dogs were at a significantly lower risk for glaucoma, compared with other breeds. Eyes with IOL placement were at a significantly lower risk for glaucoma, compared with eyes without IOL placement. Eyes with hypermature cataracts were at a significantly higher risk for glaucoma, compared with eyes with mature or immature cataracts. CONCLUSIONS AND CLINICAL RELEVANCE: Multiple factors appear to contribute to the onset of glaucoma in dogs after cataract surgery. Complications prohibiting IOL placement during cataract surgery may lead to a high risk of glaucoma development. PMID- 10844972 TI - Risk factors for enterolithiasis among horses in Texas. AB - OBJECTIVE: To identify risk factors for enterolithiasis in horses. DESIGN: Matched case-control study. ANIMALS: 26 horses with enteroliths, 104 horses with other causes of colic that underwent surgery (52 horses, surgical control group) or were treated medically (52 horses, nonsurgical control group). PROCEDURE: Medical records were reviewed for horses with enteroliths and control horses. Information collected included signalment, anamnesis, and findings on physical examination and clinicopathologic testing at admission. Horses with enteroliths and control horses were compared by means of conditional logistic regression to identify factors associated with enterolithiasis. RESULTS: Horses that were fed alfalfa hay, spent < or = 50% of time outdoors, or were Arabian or miniature breeds had an increased risk of developing enteroliths. Horses with enteroliths were more likely to have been hyperbilirubinemic and to have had clinical signs > 12 hours prior to admission. CONCLUSIONS AND CLINICAL RELEVANCE: Breed and diet appear to influence the risk of enterolithiasis; other management factors also may influence development of enteroliths. Duration of clinical signs may be longer and signs may be less severe among horses with enteroliths, compared with horses with other causes of colic. PMID- 10844973 TI - Pythiosis with bone lesions in a pregnant mare. AB - A 9-year-old pregnant mare was referred for evaluation of a nonhealing wound of 8 weeks' duration on the lateral aspect of the left forelimb. A soft tissue mass encircled the proximal two thirds of the metacarpus; radiography revealed a moderate periosteal reaction affecting metacarpal bone i.v. Histologic and immunohistochemical examinations revealed eosinophilic granulomatous inflammation and Pythium sp in the soft tissues. The mare was treated for 12 days with antimicrobials, medicated wound dressings, debridement, and i.v. administration of sodium iodide; radiography revealed progression of the bone lesions. The mare was treated by regional arterial perfusion with miconazole and excision of affected soft tissues and the distal two thirds of metacarpal bone i.v. The mare recovered without complications and gave birth to a healthy foal. Regional perfusion of antifungal agents provides high concentrations in soft and osseous tissues and permits use of low dosages of agents administered by other routes, which reduces cost, adverse effects, and teratogenic effects. PMID- 10844974 TI - Arthroscopic removal of patellar fracture fragments in horses: five cases (1989 1998). AB - OBJECTIVE: To evaluate the use of arthroscopy as the primary method for removal of large patellar fracture fragments. DESIGN: Retrospective study. ANIMALS: 5 performance horses of various breeds with patellar fractures. PROCEDURE: Clinical signs of lameness, external evidence of injury, and radiography were used to diagnose and determine fracture orientation. Arthroscopy of the stifle joint was performed on the affected limb with the horse positioned in dorsal recumbency and under general anesthesia. Progress after surgery was determined by evaluating medical records and via telephone conversations with owners. RESULTS: 4 of 5 horses had fractures of the medial aspect of the patella and 1 horse had a fracture of the lateral aspect of the patella. There were no postoperative complications with the joint or the arthroscopic portal incisions. Recovery periods ranged from 3 to 5 months. All horses recovered completely from surgery, and performed at the same or higher level of competition as before arthroscopy. CONCLUSIONS AND CLINICAL RELEVANCE: Femoropatellar joint arthroscopy is a favorable means by which evaluation of the stifle joint and removal of large fracture fragments can be achieved with negligible postoperative complications. PMID- 10844976 TI - Thermoluminescence (TL) from photosynthetic materials. PMID- 10844975 TI - Lifetime reproductive and financial performance of female swine. AB - OBJECTIVE: To evaluate reproductive and financial performance for commercial swine herds grouped on the basis of pattern of removal of female swine. DESIGN: Cohort study. SAMPLE POPULATION: 25 swine herds. PROCEDURES: Lifetime reproductive productivity was summarized as number of pigs weaned per herd day per mated female and as number of herd days per pig weaned per mated female. Factors associated with these 2 measures were determined by use of linear regression. Financial data from a commercial database were used to estimate maximum number of parities at removal associated with profitability. Sensitivity analysis was used to simulate how variations in daily maintenance cost and value per weaned pig would influence profitability. RESULTS: Mean number of pigs weaned per herd day per mated female was 0.054; mean number of herd days per pig weaned per mated female was 20.2. Both these measures were associated with proportion of nonproductive days during herd life, preweaning mortality rate per litter weaned, mean lifetime number of pigs born alive per litter weaned, and mean lifetime lactation duration. Maximum parity at time of removal associated with profitability ranged from 5 to 8. Daily maintenance costs per female had a greater impact on lifetime profitability than did value per weaned pig. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that lifetime reproductive and financial performance is optimized among swine herds that have higher proportions of high-parity females. PMID- 10844977 TI - Effect of environmental stress on radiation response of Saccharomyces cerevisiae. AB - We have investigated the effect of pH shock and oxidative stress (H2O2 effect) both separately and together on the response of Saccharomyces cerevisiae exposed to UV and gamma radiations for one hour. Exposure to these environmental stresses resulted in S. cerevisiae cells acquiring resistance to UV radiation. Presence of cycloheximide (a known protein synthesis inhibitor) during stress inhibited the acquired UV resistance. The increased UV resistance is apparently mediated through nucleotide excision repair as the stress exposure to rad3 mutants (defective in nucleotide excision repair) do not have any effect on UV response. Both types of stresses used probably follow the same path of induction of radioresistance as the effect of both of them is nonadditive. In the strains used in our study stress exposure does not have any significant effect on gamma radiation response. PMID- 10844978 TI - Inhibition of a protein tyrosine kinase activity in Plasmodium falciparum by chloroquine. AB - The aim of the present study was to establish the importance of phosphorylation events for parasite growth and maturation. Investigations into the cytosolic Plasmodium falciparum protein tyrosine kinase (PTK) activity revealed that there is a stage specific increase in the activity, in the order ring < trophozoite < schizont in both chloroquine sensitive (CQ-S) and chloroquine resistant (CQ-R) strains (p < 0.05). Our data also show that in vivo conversion of the schizont stage to ring stage via release of merozoites is associated with a decrease in PTK activity. Piceatannol, a specific inhibitor of PTK inhibited the activity in both the CQ-S and CQ-R strains of the parasites. The presence of low levels of chloroquine (CQ) inhibited the cytosolic PTK activity in a dose dependent manner (IC50 = 45 mumoles or 23 micrograms/ml) in CQ-S strains. The effect of varying concentration of CQ on the kinetics of peptide phosphorylation reveal that CQ was a competitive inhibitor of PTK with respect to peptide substrate and non competitive with respect to ATP indicating that CQ inhibits PTK activity by binding with protein substrate binding site. These data thus suggests that maturation of malaria parasite may be due to this cellular PTK and its inhibition by CQ could provide a hypothesis to explain its antimalarial activity and efficacy. PMID- 10844979 TI - Pathways of glucose catabolism in procyclic Trypanosoma congolense. AB - Studies of respiration on glucose in procyclic Trypanosoma congolense in the presence of rotenone, antimycin, cyanide, salicylhydroxamic acid and malonate have indicated the presence of NADH dehydrogenase, cytochrome b-c1, cytochrome aa3, trypanosome alternate oxidase and NADH fumarate reductase/succinate dehydrogenase pathway that contributes electrons to coenzyme Q of the respiratory chain. The rotenone sensitive NADH dehydrogenase, the trypanosome alternate oxidase, and cytochrome aa3 accounted for 24.5 +/- 6.5, 36.2 +/- 4.2 and 54.1 +/- 5.5% respectively of the total respiration. Activities of lactate dehydrogenase, NAD(+)-linked malic enzyme and pyruvate kinase were less than 6 nanomoles/min/mg protein suggesting that they play a minor role in energy metabolism of the parasite. Phosphoenolpyruvate carboxykinase, pyruvate dehydrogenase, succinate dehydrogenase, NADP(+)-linked malic enzyme, NADH fumarate reductase, malate dehydrogenase, and alpha-ketoglutarate dehydrogenase and glycerol kinase on the other hand had specific activities greater than 60 nanomoles/min/mg protein. These enzyme activities could account for the production of pyruvate, acetate, succinate and glycerol. The results further show that the amount of glycerol produced was 35-48% of the combined total of pyruvate, acetate and succinate produced. It is apparent that some of the glycerol 3-phosphate produced in glycolysis in the presence of salicylhydroxamic acid is dephosphorylated to form glycerol while the rest is oxidised via cytochrome aa3 to form acetate, succinate and pyruvate. PMID- 10844980 TI - DNA typing and microsatellite associated sequence amplification (MASA) in detecting loss or gain of alleles. AB - Constant exposure of genomic DNA to a variety of damaging agents including radiations poses a major challenge for developing an experimental approach for monitoring such damages. No single approach can be used to address this issue. In the present article, we have provided examples of DNA typing and believe that various approaches may enable to uncover not only loss or gain(s) of the alleles but also reveal subtle changes in the genome. Similarly, our genome analysis of the animal systems provides a method to assess the effects of radiation on domestic animals. PMID- 10844981 TI - In vitro effects of low-level, low-frequency electromagnetic fields on DNA damage in human leucocytes by comet assay. AB - The sources for the effects of electromagnetic fields (EMFs) have been traced to time-varying as well as steady electric and magnetic fields, both at low and high to ultra high frequencies. Of these, the effects of low-frequency (50/60 HZ) magnetic fields, directly related to time-varying currents, are of particular interest as exposure to some fields may be commonly experienced. In the present study, investigations have been carried out at low-level (mT) and low-frequency (50 Hz) electromagnetic fields in healthy human volunteers. Their peripheral blood samples were exposed to 5 doses of electromagnetic fields (2,3,5,7 and 10mT at 50 Hz) and analysed by comet assay. The results were compared to those obtained from unexposed samples from the same subjects. 50 cells per treatment per individual were scored for comet-tail length which is an estimate of DNA damage. Data from observations among males were pooled for each flux density for analysis. At each flux density, with one exception, there was a significant increase in the DNA damage from the control value. When compared with a similar study on females carried out by us earlier, the DNA damage level was significantly higher in the females as compared to the males for each flux density. PMID- 10844982 TI - Effect of low voltage on electrical properties of eye lens. AB - In the present study, the Cole-cole plot of lens tissue has been drawn using AC impedance system (EG and G PARC Model 318) in the frequency range 10 mHz to 10 Hz at low voltage stress. The impedus locus between real part (Z') and imaginary part (Z") of complex impedance of lens was examined. Results showed that the extracellular resistance (Re), distribution factor (alpha) and depressed angle (theta) were significantly varied at experimental low voltages. An attempt has been made to explain the electrical data of voltage-tissue interaction on the basis of solid state biophysics. PMID- 10844983 TI - Microwave absorption in oligomers of ethylene glycol. AB - The dielectric properties of biologically and pharmaceutically important low molecular weight ethylene glycols H(-OCH2CH2-)n -OH (n = 1,2,4,6) were investigated to clarify the effect of chain length on the dielectric properties. The measurement of dielectric constant and dielectric loss was carried out over the frequency range 200 MHz to 20 GHz at temperatures of 25 degrees C to 55 degrees C. It is found that in these molecules microwave dielectric losses are significant. The dispersion behaviour of these molecules can be represented by Cole-Cole equation. The dielectric properties of these homologous ethylene glycols are discussed in terms of the effects of chain length and intermolecular hydrogen bonds regarding the molecular conformations. These wide frequency range dielectric data have also been discussed in view of the suitable selection of the oligomer of ethylene glycol for cosmetic preparations and other pharmaceutical applications with the intention of protection of the skin from weak microwave radiations present in the surrounding environment. These systematic microwave dielectric data with frequency and temperature variation are not available and are provided in this paper. PMID- 10844984 TI - Monte Carlo simulation of photon scattering in biological tissue models. AB - Monte Carlo simulation of photon scattering, with and without abnormal tissue placed at various locations in the rectangular, semi-circular and semi-elliptical tissue models, has been carried out. The absorption coefficient of the tissue considered as abnormal is high and its scattering coefficient low compared to that of the control tissue. The placement of the abnormality at various locations within the models affects the transmission and surface emission of photons at various locations. The scattered photons originating from deeper layers make the maximum contribution at farther distances from the beam entry point. The contribution of various layers to photon scattering provides valuable data on variability of internal composition. Introduction. PMID- 10844985 TI - Effect of amplitude modulated RF radiation on calcium ion efflux and ODC activity in chronically exposed rat brain. AB - The effect of exposing rats to amplitude modulated radiofrequency radiation (112 MHz modulated to 16 Hz) during development and growth has been examined. Wistar rats (35 days old) when exposed at above frequency at the power level 1.0 mW/cm2 (SAR, 0.75 W/kg) for 35 days showed enhanced ornithine decarboxylase activity and Ca2+ efflux in brain indicating potential health hazards due to exposure. PMID- 10844986 TI - Magnetic resonance imaging: bioeffects and safety concerns. AB - Magnetic resonance imaging (MRI) is the state-of-the-art noninvasive imaging modality in clinical diagnosis. During MRI examination, the patient is exposed to three different forms of electromagnetic radiation: (i) a static magnetic field, (ii) gradient magnetic fields, and (iii) radiofrequency (RF) fields. Each of these may cause significant adverse bioeffects if applied at sufficiently high exposure levels. This article describes in some detail the areas of health concern for both the patient and the health practitioner with respect to the use of clinical MRI, in addition to describing the potential bioeffects of electromagnetic radiations used in this sophisticated imaging modality. PMID- 10844987 TI - Effects of extremely low frequency electromagnetic fields on health. AB - This paper gives a brief review of the physical interaction and bio-effects of exposure to extremely low frequency (ELF) electromagnetic fields (EMF) along with guidelines on limits of exposure to 50/60 Hz electric and magnetic fields. PMID- 10844988 TI - Issues in electromagnetic field-biointeractions. AB - Low level electromagnetic fields have been found to produce a variety of biological effects, though the mechanism of such interaction is still not completely understood. Cell membrane of the brain is a critical structure perceiving the action of microwaves, which has received greater attention in the recent past. The interactions of EMF with the living cells are considered as stochastic resonance, cooperative effects, non-equilibrium thermodynamic process and non-linear interactions. The living cells derive the energy from noise and pumps it into the modes of excitation at the driving frequency of an electromagnetic wave which give sufficient amplification of the signal and increase the signal to noise ratio. The non-linear mechanism plays their main role in the process of transmembrane coupling of the signal to the cytoplasm. The criteria for safe exposure limits of electromagnetic field to humans is also discussed. PMID- 10844989 TI - Static magnetic field as biological modifier: a study on temperature dependent influence. AB - Magnetic fields seemingly alter a number of physiological indicators in intact animals and influence cellular metabolism. We have studied the magnetic field effects on the membrane and receptors of the reticulo-endothelial cells of bone marrow which are mainly responsible for the phagocytic activity of nanocolloid particles of human serum albumin tagged with Tc99m. A series of experiments carried out on immobilized mice exposed to a static 1.4 T SMF for 60 min at 27 degrees C or 37 degrees C body temperature showed an increased phagocytic activity at 37 degrees C and decreased activity at 27 degrees C. PMID- 10844990 TI - Effects of low level ionising radiations. PMID- 10844991 TI - Electromagnetic field in and around the campus of Jawaharlal Nehru University, New Delhi. AB - We have surveyed the levels of magnetic field in the open environment around the high-tension lines as well as in the domestic and laboratory environment in the JNU campus. Measurements were made in the direction of most likely access, and from the surface of the appliances. The data so obtained are analyzed and compared with some quoted in the literature. It is hoped that the results will be useful for epidemiological studies in determining the criteria for safe human exposure. PMID- 10844992 TI - A perspective of biological supramolecular electron transfer. AB - Electron transfer is an essential activity in biological systems. The migrating electron originates from water-oxygen in photosynthesis and reverts to dioxygen in respiration. In this cycle two metal porphyrin complexes possessing circular conjugated system and macrocyclic pi-clouds, chlorophyll and heme, play a decisive role in mobilising electrons for travel over biological structures as extraneous electrons. Transport of electrons within proteins (as in cytochromes) and within DNA (during oxidative damage and repair) is known to occur. Initial evaluations did not favour formation of semiconducting pathways of delocalized electrons of the peptide bonds in proteins and of the bases in nucleic acids. Direct measurement of conductivity of bulk material and quantum chemical calculations of their polymeric structures also did not support electron transfer in both proteins and nucleic acids. New experimental approaches have revived interest in the process of charge transfer through DNA duplex. The fluorescence on photo-excitation of Ru-complex was found to be quenched by Rh-complex, when both were tethered to DNA and intercalated in the base stack. Similar experiments showed that damage to G-bases and repair of T-T dimers in DNA can occur by possible long range electron transfer through the base stack. The novelty of this phenomenon prompted the apt name, "chemistry at a distance". Based on experiments with ruthenium modified proteins, intramolecular electron transfer in proteins is now proposed to use pathways that include C-C sigma-bonds and surprisingly hydrogen bonds which remained out of favour for a long time. In support of this, some experimental evidence is now available showing that hydrogen bond-bridges facilitate transfer of electrons between metal-porphyrin complexes. By molecular orbital calculations over 20 years ago we found that "delocalization of an extraneous electron is pronounced when it enters low-lying virtual orbitals of the electronic structures of peptide units linked by hydrogen bonds". This review focuses on supramolecular electron transfer pathways that can emerge on interlinking by hydrogen bonds and metal coordination of some unnoticed structures with pi-clouds in proteins and nucleic acids, potentially useful in catalysis and energy missions. PMID- 10844993 TI - Significance of charge on lysine residue of ovine luteinizing hormone on immunological and biological properties of the hormone. AB - In order to understand the significance of positive charge of lysine residues of ovine luteinizing hormone (oLH) on immunological and biological activity, the epsilon-NH2 group(s) of ovine LH were sequentially modified with 2-iminothiolane (2IT) that preserves the positive charge of the lysine while the overall charge of the hormone remains unchanged. These studies have also been compared with the oLH modified by N-succinimidyl 3-(2 pyridyldithio) propionate (SPDP) and succinimidyl 6-[3-(2-pyridyldithio)propionamido]hexanoate (LC-SPDP) that abolish positive charge of lysine residues. The modification primarily occurs in the alpha-subunit. Sequential modification led to progressive reduction in receptor binding and immunological activities. However, the steroidogenic activity was substantially retained. The immunoreactivity and receptor binding properties of 2IT modified oLH (oLH-2IT) were less affected when compared to SPDP (oLH-SPDP) or LC-SPDP (oLH-LC-SPDP) modified derivatives suggesting that increase in hydrophobic carbon chain in oLH-LC-SPDP molecule resulted in drastic inhibition in immunological and biological properties. But the steroidogenic potential of oLH-2IT, oLH-LC-SPDP or oLH-SPDP was relatively comparable. This suggests that a single -NH2 group modification with 2IT would generate the site in the hormone for conjugation to the toxin/carrier proteins that may retain better immunological and biological activity compared to that of SPDP or LC-SPDP modified oLH. PMID- 10844994 TI - The role of LFA-1, Mac-1, ICAM-1 and Ia in the induction of Th-2 type of immune response in spleen during murine syngeneic pregnancy. AB - Murine syngeneic pregnancy is characterized by the transient splenomegaly at mid gestation. Recent studies from our laboratory have indicated the initiation of T cell dependent B-cell response in the spleen during early pregnancy (Hegde and Nainan 1998). Present studies were carried out to understand the role of cell adhesion and MHC class II (Ia) molecules in the induction of Th-2 type of response in the spleen of pregnant mouse. Immunochemical localization of ICAM-1, LFA-1, Mac-1 and Ia in spleen have been carried out at different stages of pregnancy and formation of cell clusters and natural cell adhesion assay with splenocytes were carried out on day 1 (D1) pregnancy and compared with control. Upregulation of ICAM-1, LFA-1, Mac-1 and Ia was observed during early pregnancy. This coincided with the formation of germinal centers (GC) and Th2 type of interleukins in spleen as reported earlier. Increased expression of cell adhesion and Ia molecules during early pregnancy provides additional evidence for the systemic shift to Th2 type of immune response in syngeneic murine pregnancy. PMID- 10844995 TI - Metal ion specificity in anaesthetic induced increase in the rate of monensin and nigericin mediated H+/M+ exchange across phospholipid vesicular membranes. AB - From a study of the decay of the pH difference across vesicular membranes (delta pH) it has been possible to show that H+ and alkali metal ion (M+) concentration gradients across bilayer membranes (which are responsible for driving important biochemical processes) can be selectively perturbed by anaesthetics such as chloroform and benzyl alcohol by combining them with a suitable exchange ionophore. On adding the anaesthetic to the membrane in an environment containing metal ions M+ = K+, the rate of delta pH decay by H+/M+ exchange increases by a larger factor or by a smaller factor (when compared to that in a membrane environment with M+ = Na+) depending on whether the exchange ionophore chosen is monensin or nigericin. A rational explanation of this "metal ion specificity" can be given using the exchange ionophore mediated ion transport scheme in which the equilibrations at the "interfaces" are fast compared to the "translocation equilibration" between the species in the two layers of the membrane. The following three factors are responsible for the observed "specificity": On adding the anaesthetic (i) translocation rate constants increase, (ii) the concentrations of the M+ bound ionophores increase at the expense of H+ bound ionophores. (iii) Under our experimental conditions the rate determining species are the complexes monensin-K (Mon-K) and nigericin-H (Nig-H) for M+ = K+ whereas they are monensin-H (Mon-H) and nigericin-Na (Nig-Na) for M+ = Na+. Possible anaesthetic induced membrane perturbations contributing to the above mentioned changes in the membrane are (A), the loosening of the membrane structure and (B), an associated increase in the membrane hydration (and membrane dielectric constant). An analysis of the consequent changes in the various transport step shows the following: (a), The anaesthetic induced changes in the translocation rates of electrically charged species are not relevant in the explanation of the observed changes in the delta pH decay rates. (b), Changes in the rates of fast equilibria at the interface contribute to changes in KH and KM. (c), A suggestion made in the literature, that a significant interaction between the dipole moment of the monensin-K complex and the membrane slows down its translocation, is not valid. (d), The ability to explain rationally all the delta pH decay data confirms the validity of the transport scheme used. In our experiments delta pH across the vesicular membrane was created by pH jump coming from a temperature jump. PMID- 10844996 TI - Conformations of hydrophobic peptides in trifluoroethanol, water and in solid state: a circular dichroism and Fourier Transform Infrared study. AB - The conformations of peptides corresponding to KLLIALVLCFLPLAALG have been examined in trifluoroethanol (TFE), aqueous medium by circular dichroism spectroscopy and in the solid state by Fourier Transform Infra Red Spectroscopy (FTIR). The 17-residue parent peptide and peptides corresponding to shorter segments LVLCFLPLAALG and CFLPLAALG showed preference for helical conformation in TFE. Even the shorter hydrophobic peptides corresponding to KLLIA and LVL showed propensity for beta-turn conformations in TFE. However, peptides corresponding to the relatively polar segment FLPLAALG were unordered in TFE. In water, peptides that showed ordered conformation in TFE preferred beta-conformation. In solid state, FTIR spectra indicated that the hydrophobic peptides adopt beta-structures with extensive hydrogen bonded network in the solid-state. The hydrophobic core segment thus appears to dictate the conformational propensity of the peptide. PMID- 10844997 TI - Cloning, expression and purification of the DNA binding domain of RFX protein. AB - The RFX DNA binding domain (DBD) is a novel highly conserved motif belonging to a large number of dimer DNA binding proteins which have diverse regulatory functions in eukaryotic organisms. To characterize this novel motif, a 78mer polypeptide corresponding to the DBD of human hRFX (hrfX1/DBD), a prototypical member of the RFX family has been cloned and overproduced in Escherichia coli. A purification procedure using cation exchange chromatography has also been developed. PMID- 10844998 TI - UDP-galactose 4-epimerase from Kluyveromyces fragilis: equilibrium unfolding studies. AB - UDP-galactose 4-epimerase from yeast (Kluyveromyces fragilis) is a homodimer of total molecular mass 150 kDa having possibly one mole of NAD/dimer acting as a cofactor. The molecule could be dissociated and denatured by 8 M urea at pH 7.0 and could be functionally reconstituted after dilution with buffer having extraneous NAD. The unfolded and refolded equilibrium intermediates of the enzyme between 0-8 M urea have been characterized in terms of catalytic activity, NADH like characteristic coenzyme fluorescence, interaction with extrinsic fluorescence probe 1-anilino 8-naphthelene sulphonic acid (ANS), far UV circular dichroism spectra, fluorescence emission spectra of aromatic residues and subunit dissociation. While denaturation monitored by parameters associated with active site region e.g. inactivation and coenzyme fluorescence, were found to be cooperative having delta G between -8.8 to -4.4 kcals/mole, the overall denaturation process in terms of secondary and tertiary structure was however continuous without having a transition point. At 3 M urea a stable dimeric apoenzyme was formed having 65% of native secondary structure which was dissociated to monomer at 6 M urea with 12% of the said structure. The unfolding and refolding pathways involved identical structures except near the final stage of refolding where catalytic activity reappeared. PMID- 10844999 TI - Inhibition of thymidylate synthase by pergularinine, tylophorinidine and deoxytubulosine. AB - The activity of thymidylate synthase (TS) purified in our laboratory from Lactobacillus leichmannii was inhibited by pergularinine (PGL) and tylophorinidine (TPD) and deoxytubulosine (DTB) isolated from the Indian medicinal plants Pergularia pallida and Alangium lamarckii respectively. Cytotoxicity studies showed that cell growth of L. leichmannii was inhibited (IC50 = 40-45 microM) by all the three alkaloids, the concentrations > 80-90 microM resulting in complete loss of the enzyme activity. Ki values of the enzyme calculated from Lineweaver-Burk and Dixon plots for PGL, TPD and DTB were 10 x 10(-6) M, 9 x 10(-6) M and 7 x 10(-6) M respectively. These are typed as 'non competitive' inhibitors of TS. All the three alkaloids inhibited (IC50 = 50 microM) the elevated TS activity of leukocytes in cancer patients with clinically diagnosed chronic myelocytic leukemia (n = 10), acute lymphocytic leukemia (n = 8) and metastatic solid tumours (n = 3). PMID- 10845000 TI - Immobilization of Amaranthus leaf oxalate oxidase on alkylamine glass. AB - A membrane bound oxalate oxidase from leaves of Amaranthus spionsus has been partially purified and immobilized on alkylamine glass with a yield of 9.2 mg protein/g support. The enzyme retained 99.4% of initial activity of free enzyme after immobilization. There was no change in the optimum pH (3.5) and Vmax but the temperature for maximum activity was slightly decreased (35 degrees C) and energy of activation (Ea) and Km for oxalate were increased after immobilization. The immobilized enzyme preparation was stable for 6 months, when stored in distilled water at 4 degrees C. Presence of Cl- did not affect the activity of immobilized enzyme. PMID- 10845001 TI - [Current morphological classification of chronic liver inflammation: its merits and problems]. PMID- 10845002 TI - [Electron microscopy of viral diseases in the course of chronic viral inflammation of the liver]. PMID- 10845003 TI - [Lymphadenoplasia as a morphological feature of chronic viral inflammation of the liver]. PMID- 10845004 TI - [Steps in the diagnosis of chronic inflammation of the liver]. PMID- 10845005 TI - [The organization of and the equipment in the pathomorphological laboratory and the principles of dealing with the materials of histopathological and cytological research]. PMID- 10845006 TI - Changing assumptions about the mechanism of corneal transplant rejection. PMID- 10845007 TI - Unnatural injuries. PMID- 10845008 TI - Radiotherapy for choroidal neovascularisation of age-related macular degeneration: a fresh perspective. PMID- 10845009 TI - Proton beam irradiation of subfoveal choroidal neovascularisation in age-related macular degeneration. AB - PURPOSE: To assess the safety and potential toxicity of proton beam radiation in the treatment of subfoveal choroidal neovascular membrane (CNVM) due to age related manner degeneration (ARMD) in a prospective, non-randomised study. METHODS: Forty-eight eyes of 46 consecutive patients with subfoveal CNVM due to ARMD, not amenable to laser photocoagulation, were treated prospectively with a single proton beam exposure. Two dose regimens were evaluated: 8 CGE (Cobalt Gray Equivalent) and 14 CGE. Patients were followed for an average of 22.1 months after proton beam treatment. RESULTS: At the 12 month follow-up, 44% of eyes in the 8 CGE group and 75% of the eyes in the 14 CGE group had stabilized or improved visual acuity. Complex size in the 8 CGE group as measured on standard fluorescein angiography (FA), decreased or had no change initially but showed less effect over time, while the eyes treated with 14 CGE maintained decreased leakage over the follow-up period of 12 months. However, 11 eyes in the 14 CGE group experienced radiation retinopathy, with the onset between 3 and 30 months. Seven of these 11 eyes have demonstrated some visual loss but only 1 eye developed severe visual loss at 15 months after proton treatment. CONCLUSIONS: To date, 14 CGE has suggested a favourable influence on visual function and growth inhibition of CNVM. Proton beam irradiation appears to inhibit CNVM growth. The 14 CGE dose regimen appears to have a longer effect of CNVM growth than does 8 CGE, with overall stabilisation of visual function and growth inhibition. Radiation retinopathy has developed over time, but severe visual loss has been limited. On the basis of the incidence of radiation retinopathy, adjustments in the total radiation dosage and/or fractionation of the dosage should be considered. PMID- 10845010 TI - Effects of scleral buckling on the retrobulbar haemodynamic changes. AB - PURPOSE: This study was planned to investigate blood flow changes due to scleral buckling surgery. In addition the effects on these changes of factors related to patient characteristics and operative technique were studied. METHODS: Central retinal artery (CRA) and ophthalmic artery (OA) blood flow velocities were studied with colour Doppler ultrasonography after scleral buckling surgery in 25 patients with unilateral rhegmatogenous retinal detachment. The effects of the patient's age, referral time, aetiological factors, operative technique, cryotherapy width and buckling distance on the haemodynamic changes were noted. RESULTS: Buckling surgery reduces the blood flow velocities in the CRA, but affects OA blood flow less. Encircling is found to be responsible and it is greater in patients with anatomical success. Other factors do not have any significant effect on these changes. CONCLUSIONS: Good anatomical and functional results can be achieved despite these haemodynamic changes. It must be kept in mind that these changes may cause complications in some patients and the surgery must be minimized. PMID- 10845012 TI - Complications of extracapsular cataract surgery in chronic renal failure patients. AB - PURPOSE: In chronic renal failure (CRF) patients the risk of per- and post operative complications in cataract surgery is high. The most frequent complications observed in these patients and the prevalence of cataract were studied. METHODS: Eighty-two CRF patients who had undergone ophthalmic examination between December 1996 and April 1998 at Baskent University, Department of Ophthalmology, were retrospectively analysed. Of 82 patients, 18 eyes of 14 CRF patients between 37 and 79 years of age underwent cataract extraction. Planned extracapsular cataract extraction was performed in all patients and all but one had posterior chamber intraocular lens implantation. In the post-operative period, scrapings of the corneal infiltrates were cultured and smears were examined, and the conjunctiva was swabbed and cultured also. RESULTS: Per-operative hypotony and scleral collapse occurred in 1 patient, and another had haemorrhage in the anterior chamber. Post-operatively, suture infiltration was observed in 10 eyes. These lesions disappeared after topical steroid and antibiotic treatment. CONCLUSIONS: Patients with CRF require special surgical considerations since they face both general surgical problems and others that arise due to their disease. PMID- 10845011 TI - Linkage analysis in an autosomal dominant 'zonular nuclear pulverulent' congenital cataract, mapped to chromosome 13q11-13. AB - PURPOSE: To determine by linkage analysis the chromosomal locus responsible for autosomal dominant congenital cataract (ADCC) in a four-generation Welsh pedigree. METHODS: The family pedigree was traced through two members of the family attending the hospital for treatment of their cataracts. Twenty-five members of the family were examined ophthalmologically and blood was collected with consent for genetic linkage analysis. RESULTS: Fifteen members of this family were known to have bilateral congenital cataracts, of whom 11 had a cataract extraction prior to examination. The youngest member of the pedigree was 5 years old and the oldest was 78 years old. Four children, unoperated at the time of this study, had a phenotypically identical morphology of their zonular pulverulent congenital cataracts. The known loci for congenital cataract were excluded. Significant lod scores for markers in the 13q11-13 region were detected with a Zmax of 3.59 D13S1236 (theta = 0.00). CONCLUSIONS: We report linkage of an ADCC of the zonular pulverulent type to chromosome 13q11-13 in a four-generation family. PMID- 10845013 TI - Vertical diplopia following local anaesthetic cataract surgery: predominantly a left eye problem? AB - PURPOSE: Vertical diplopia is an uncommon but disappointing complication of otherwise successful local anaesthetic cataract surgery. We studied strabismus patterns in a group of such patients to identify the nature and extent of extraocular muscle involvement. METHODS: A retrospective review identified 15 cases of vertical diplopia following local anaesthetic cataract surgery between July 1994 and January 1998. Peribulbar anaesthesia was used in all cases and given by right-handed professionals. RESULTS: All cases had otherwise successful cataract surgery (mean age 80.5 years; median pre-operative VA 6/18; median post operative VA 6/9). The mean level of vertical diplopia was 7.2 prism dioptres (PD) in the primary position (range 2-25 PD). The left inferior rectus (IR) was paretic in 6 cases and restricted in 5 cases. The left superior rectus (SR) was not affected in any of the cases. The right IR was restricted in a single case. The right SR was paretic in 2 cases and restricted in a single case. None of the cases had clinical involvement of the oblique muscles. Eleven of the cases were managed successfully with prisms. Two of the cases required strabismus surgery. CONCLUSIONS: The incidence of left eye extraocular muscle involvement was greater than right eye involvement, although this did not reach statistical significance (73% vs 27%; p = 0.075). This may be due to the more difficult access of right handed individuals giving left eye peribulbar injections with the needle tract being directed more closely to the muscle cone. The IR muscle is more commonly affected than the SR (80% vs 20%; p = 0.019). An equal incidence of paretic and restricted rectus muscle pathology was found in this study (53% vs 47%; p = 0.818). The exact aetiology of muscle injury is unknown but could be due to direct muscle or nerve trauma, anaesthetic toxicity, periocular haemorrhage or a combination of these. PMID- 10845014 TI - Post-operative changes in the capsulorhexis aperture: a prospective, randomised comparison between loop and plate haptic silicone intraocular lenses. AB - PURPOSE: There is disagreement regarding whether the capsulorhexis aperture always decreases or may increase post-operatively. The aim of this study was to settle the controversy concerning loop haptic silicone lenses and to learn more of the dynamics of the capsulorhexis relating to plate haptic lenses. METHODS: We performed a prospective randomised study comparing the post-operative changes in capsulorhexis aperture in two groups of eyes implanted with either plate or loop haptic silicone intraocular lenses. All the surgery was performed, at a teaching hospital in the United Kingdom, by a single surgeon, using a standard technique of phacoemulsification. Patients were reviewed at 2 weeks and 6 months post operatively. Digital retroillumination images of the anterior segment were captured. The area of the capsulorhexis aperture was determined by manually detecting its edge on a computer monitor. RESULTS: Forty-eight cases were randomised. The groups were comparable for demographic variables and mean initial aperture size (p > 0.05). There was an 8.4% mean decrease in aperture size for the loop haptics, contrasting with 4.5% expansion for the plate haptics (p < 0.05). Sixty-five per cent of patients with the plate haptic underwent enlargement of the aperture, contrasting with 25% for the loop haptic lens (p < 0.05). CONCLUSIONS: Silicone lenses with plate haptics undergo expansion of the capsulorhexis aperture more frequently than those with Prolene loop haptics. PMID- 10845015 TI - The phacoemulsification learning curve: per-operative complications in the first 3000 cases of an experienced surgeon. AB - PURPOSE: To assess the per-operative complications occurring during the first 3000 phacoemulsification cases performed by an experienced consultant surgeon. METHODS: A prospective analysis of 3000 consecutive cases performed without supervision between November 1992 and November 1998 was carried out. Data recorded for each case included details of per-operative complications, pre operative best corrected visual acuity, nuclear density, history of previous pars plana vitrectomy, and whether phacoemulsification was performed as part of a phacotrabeculectomy procedure. RESULTS: The overall rate of vitreous loss was 1.3%. Nuclear fragments were lost to the vitreous in 6 cases (0.2%). The initial rate of vitreous loss was 4.0% in the first 300 cases falling to 0.7% in the last 300 cases. Capsulorhexis failure was the commonest per-operative complication observed, but the risk of subsequent posterior capsule rupture fell significantly from 9 of 45 (20.0%) in the first 100 cases to 1 of 49 (2.0%) in the next 2000 cases (p = 0.0061, Fisher's exact test). There was a significant increase in risk with denser cataracts, especially for capsulorhexis failure, rising to over 35% in the densest cases. The increases in posterior capsule rupture and vitreous loss were less dramatic but nonetheless very significant. There was no significant increase in the risk of per-operative complications with phacotrabeculectomy, and no increased risk in patients who had previously undergone pars plana vitrectomy. Posterior capsule rupture occurred in 22 of 612 (3.6%) local anaesthesia cases compared with 31 of 2269 (1.4%) topical anaesthesia cases. Per-operative best corrected visual acuity of 6/9 or better was recorded in 2.0% of the first 1000 cases compared with 13.9% of the last 1000 cases. CONCLUSIONS: (1) Per-operative surgical risks could be reduced to low levels during the learning curve, but complications continued to occur at a low frequency. (2) The risk of per-operative complications was not significantly elevated in previously vitrectomised eyes. (3) Nuclear density correlated significantly with per-operative complication risk. (4) The visual threshold for cataract surgery fell dramatically with increasing experience of phacoemulsification. (5) Topical anaesthesia was not associated with an increased risk of per-operative complications. PMID- 10845016 TI - Corneoscleroplasty with maintenance of the angle in two cases of extensive corneoscleral disease. AB - PURPOSE: We report a 2 year follow-up in two patients after corneoscleroplasty. METHOD: Using lamellar corneoscleral dissection to maintain the drainage angle and its function, we performed a 14 mm allograft corneoscleroplasty in 2 eyes. For surgical treatment in both cases a 9.5 mm corneal button was excised from the recipient after peritomy and scleral lamellar preparation up to 14 mm. A 14 mm donor button was inserted and held in place with multiple Prolene sutures. One eye presented with a large perforating corneal ulcer after herpetic keratitis in a patient with recurrent rheumatoid uveitis associated with rubeosis iridis. The second eye had had a penetrating keratoplasty for keratoconus 30 years previously and presented with decompensating keratoglobus. Immune suppression was performed with systemic cyclosporin A and additional steroids when required. RESULTS: Both patients had a clear graft at the last follow-up visit and visual acuity was improved to a best corrected visual acuity of 0.6. Intraocular pressure in the keratoglobus eye was maintained at 6 mmHg without treatment, whereas the second case required continuing treatment with systemic acetazolamide because of neovascular glaucoma. The anterior chamber angles remained open in both patients. Contact lenses were helpful in the prevention of epithelial irregularities and defects. Both patients had an episode of immunological graft reaction which was reversed by immunosuppressive treatment. Phacoemulsification with intraocular lens implantation, which was performed 2 years after transplantation in the keratoglobus eye, did not affect the graft clarity or cause rejection episodes. CONCLUSION: Our results using corneoscleroplasty have been encouraging in severe destructive corneal disease. PMID- 10845017 TI - Histocompatibility Y antigen compatibility and allograft rejection in corneal transplantation. AB - PURPOSE: To assess the effects of histocompatibility Y (H-Y) antigen matching on the rate of corneal allograft rejection after penetrating keratoplasty (PKP). METHODS: We retrospectively investigated the graft survival rate and rejection free graft survival rate after PKP in 396 eyes. The compatible combinations of H Y antigen included male donors and male recipients (n = 135), female donors and male recipients (n = 107), and female donors and female recipients (n = 60). Incompatible combination was from male donors and female recipients (n = 94). The eyes were classified into two groups--high-risk (168 eyes) and low-risk (228 eyes)--depending on the degree of vascularisation in the recipient corneas or a history of previous allograft rejection. Data were analysed using the Kaplan Meier life table method, the log-rank test and the Cox proportional hazards model. RESULTS: In both the high-risk and low-risk groups, the graft survival and rejection-free graft survival rates were not affected by the H-Y compatibility. The graft survival (p < 0.001) and rejection-free graft survival (p < 0.001) rates were higher in the low-risk group than in the high-risk group. High-risk PKP was associated with greater risk of graft failure (risk ratio, 2.33) and rejection (risk ratio, 2.05) than low-risk PKP. CONCLUSION: H-Y antigen matching does not influence the rate of allograft rejection after PKP. PMID- 10845018 TI - The effects of the topical administration of non-steroidal anti-inflammatory drugs on corneal epithelium and corneal sensitivity in normal subjects. AB - PURPOSE: To study the changes in the corneal epithelium and corneal sensitivity of healthy subjects after the topical administration of non-steroidal anti inflammatory drugs (NSAIDs; diclofenac, indomethacin, flurbiprofen and ketorolac) frequently used in ocular therapy. METHODS: A double-masked parallel clinical study was undertaken on 90 subjects (45 men, 45 women; Caucasian; age 21-46 years, mean +/- SD 27.1 +/- 5 years). The subjects were divided into six groups: group 1 was treated with placebo, group 2 with 0.1% diclofenac, group 3 with 0.1% indomethacin, group 4 with 0.03% flurbiprofen, group 5 with 0.5% ketorolac and group 6 with 0.4% oxybuprocaine. One eye was randomly treated with the study drug and the fellow eye was treated with placebo. The medications were instilled four times, at 5 min intervals. Assessment of the corneal epithelium was carried out by vital fluorescein stain before instillation and 5, 15, 30 and 60 min after instillation of the last drop. Subjective burning sensation was assessed by asking participants to rate burning on a scale from 0 (none) to 3 (severe). After 1 week, assessment of corneal sensitivity was carried out by the Cochet-Bonnet method, repeating the above scheme of instillation and measurement times. RESULTS: None of the study drugs, with the exception of oxybuprocaine, produced evident epithelial damage. All the drugs caused a mean burning sensation greater than the placebo. The diclofenac-treated group showed a statistically significant decrease in corneal sensitivity (p < 0.001) at the measurement carried out 15 min after instillation of the last drop and lasting up to the end of the study, when the corneal anaesthesia was similar to that induced by the topical anaesthetic treatment. No significant changes were demonstrated for the other NSAIDs when compared either with the placebo-treated eyes or with the fellow eyes. CONCLUSIONS: Despite a similar mechanism of action and analgesic activity to the other NSAIDs tested, diclofenac was able to induce a reduction in corneal sensitivity. More studies are needed to determine the mechanism of action responsible for this effect. PMID- 10845019 TI - Anterior segment indocyanine green angiography in scleral inflammation. AB - PURPOSE: To assess the feasibility of using indocyanine green angiography in scleral inflammation; to define the characteristic patterns of the anterior segment vasculature for this anterior segment disease; and to correlate the findings with those of anterior segment fluorescein angiography. METHODS: Anterior segment fluorescein and indocyanine green digital angiography were used to evaluate scleral inflammation in 3 patients with diffuse episcleritis, 2 patients with nodular episcleritis and 5 patients with nodular scleritis. Angiograms from both techniques were assessed based on the time for complete disappearance of the dye and the type of leakage. RESULTS: Both fluorescein and indocyanine green dye appeared in vessels at approximately the same time, but, whereas fluorescein had disappeared completely from vessels by the 70th second, indocyanine green was observed within vessels for up to 23 min. Leakage of fluorescein occurred in all patients with diffuse episcleritis but staining occurred in only 1 patient with nodular scleritis. No leakage of indocyanine green, or staining, occurred in patients with diffuse episcleritis. However, leakage of indocyanine green was apparent in all nodular episcleritis and scleritis patients, staining the nodules in patchy form. CONCLUSION: The longer transit time, as well as leakage and staining patterns due to its protein-binding properties, make indocyanine green angiography a potentially useful technique in the investigation of patients with scleral inflammation, and in distinguishing diffuse from nodular variants. Further studies are necessary to correlate staining patterns with clinical findings. PMID- 10845020 TI - Vitrectomy for vitreous opacification in Fuchs' heterochromic uveitis. AB - PURPOSE: To assess the role of pars plana vitrectomy (PPV) for symptomatic vitreous opacification in a series of patients with Fuchs' heterochromic uveitis (FHU). METHODS: A retrospective review was undertaken of 13 patients with FHU who underwent vitrectomy for vitreous opacification between April 1989 and December 1998. RESULTS: An improvement in visual symptoms was recorded in all patients, 9 of 13 (69%) demonstrating at least a 2 line increase in Snellen visual acuity. All but one patient attained 6/9 or better visual acuity post-operatively. Surgery was uneventful and did not appear to exacerbate any existing intraocular inflammation. CONCLUSION: From this series we conclude that PPV has an important role in the management of patients with FHU who present with symptomatic vitreous opacification. PMID- 10845021 TI - Requirements for optical services in children with microphthalmos, coloboma and microcornea in southern India. AB - PURPOSE: The aim of the study was (1) to determine the need for spectacles in children in Southern India with coloboma, microphthalmos and microcornea, (2) to describe their refractive errors and (3) to assess their needs for low vision aids (LVAs). METHODS: Children with congenital eye anomalies were recruited from special education for the blind, schools for the mentally handicapped, community based rehabilitation programmes and hospital records in Andhra Pradesh, India. All those with at least light perception vision (PL) in one eye and who had navigational vision were refracted. Those whose distance vision in their better eye improved with refraction were prescribed spectacles. Those unable to read N10 were assessed for LVAs for near. Those with distance visual acuity of < 6/18 in the better eye were assessed for telescopes to aid distance vision. RESULTS: Ninety-nine children with coloboma, microcornea or microphthalmos had functional vision. Eight unilateral cases were excluded. Ninety-one bilateral cases were refracted and assessed for LVAs. The vision in 52 children (57%) improved in the better-seeing eye by 1 or more lines of Snellen acuity with spectacles. Spectacles were prescribed most frequently for myopia associated with choroidal coloboma. After refraction, all 19 children with a visual acuity of 6/18 or better could read N10, and 43 of the 72 children (60%) with a visual acuity of < 6/18 to PL with functional vision could read N10 unaided, or with distance correction. A further 6 (8%) reached this level with magnifiers. Thirteen children (18%) were given telescopes. CONCLUSION: Children with congenital anomalies of the eye and functional vision benefit from refraction and low vision services. PMID- 10845022 TI - Antibiotics in the irrigating solutions reduce Staphylococcus epidermidis adherence to intraocular lenses. AB - PURPOSE: To investigate the effect of antibiotics in the irrigating solutions on hydrophobicity, slime production and the adherence of Staphylococcus epidermidis to intraocular lenses (IOLs). METHODS: A standard culture of S. epidermidis was incubated with a control phosphate-buffered saline (PBS) or PBS containing vancomycin (20 micrograms/ml) or gentamicin (8 micrograms/ml) or a combination of gentamicin and vancomycin (8 and 20 micrograms/ml, respectively) for 30, 60 and 120 min at 35 degrees C. The bacteria were harvested by centrifugation, and washed with PBS before incubation with IOLs for 1 h. Adhesion of bacterial cells to IOLs was determined by counting the viable cells attached to the lenses. Slime production on IOLs was measured using safranin staining. Hydrophobicity of the control cultures and cultures treated with antibiotics was assayed on the basis of the hexadecane droplet method. RESULTS: Bacterial exposure to antibiotics produced a time-dependent significant decrease in bacterial hydrophobicity and adherence to IOLs compared with the untreated control cells (p < 0.001). Hydrophobicity showed a significant correlation with adherence (r = 0.89, p < 0.001). Gentamicin was significantly more effective than vancomycin, and the synergistic combination of gentamicin and vancomycin was the most effective in reducing bacterial adherence to IOLs, hydrophobicity and slime production. CONCLUSIONS: The use of antibiotics in the irrigating solutions during cataract surgery may be useful in reducing bacterial adherence to IOLs. Further studies are needed to determine the clinical implications of these findings in reducing the incidence of post-operative endophthalmitis associated with IOL implantation. PMID- 10845023 TI - Topical vitamin E and hydrocortisone acetate treatment after photorefractive keratectomy. AB - PURPOSE: To investigate the effects of topical vitamin E and hydrocortisone acetate treatments on corneal healing response after -10.0 D photorefractive keratectomy (PRK) in rabbits. METHODS: Thirty-three New Zealand white rabbits were divided into four groups and -10 D PRK was performed under in vivo conditions. Following PRK, group 1 (n = 9) received no topical treatment and served as control. Group 2 (n = 8) received 0.1% hydrocortisone acetate ointment twice a day, group 3 (n = 8) received 1% vitamin E ointment and group 4 (n = 8) received both 0.1% hydrocortisone acetate and 1% vitamin E twice a day for a month. At the end of the third month, corneal haze was graded and the corneal hydroxyproline levels were measured, as a crude indicator of new collagen synthesis. Finally corneal samples were examined by transmission electron microscopy. RESULTS: Non-homogeneously distributed strong haze was identified in group 1 which was greater than in the other groups; haze was least in groups 2 and 4. Corneal hydroxyproline levels were found to be significantly lower in groups 2, 3 and 4 compared with the control (Student's t-test, p < 0.05). Histopathologically, the most aggressive wound healing response was detected in group 1. The corneal wound healing response of group 2 was less than that of group 1 and equal to or more than that of group 4. CONCLUSIONS: Deep corneal photoablation induces an aggressive healing response, and topical hydrocortisone acetate reduces this corneal wound healing effectively. The inhibitory effect of topical vitamin E on corneal wound healing seems to be less than that of hydrocortisone acetate, but combined treatment with these two drugs may have an additive effect in controlling corneal wound healing after PRK. PMID- 10845024 TI - Circumscribed choroidal haemangioma in a patient with Sturge Weber syndrome. PMID- 10845025 TI - An unusual presentation of a case of blue rubber bleb naevus syndrome. PMID- 10845026 TI - Mantle cell lymphoma presenting as a choroidal mass: part of the spectrum of uveal lymphoid infiltration. PMID- 10845027 TI - Spontaneous resolution of bilateral macular haemorrhage in a patient with kala azar. PMID- 10845028 TI - Squamous cell carcinoma of the conjunctiva as initial presenting sign in a patient with acquired immunodeficiency syndrome (AIDS) due to human immunodeficiency virus type-2. PMID- 10845029 TI - Scedosporium (Pseudallescheria) fungal infection of a sponge explant. PMID- 10845030 TI - Conservative management of double penetrating ocular injuries. PMID- 10845031 TI - Herpes zoster ophthalmicus presenting as contralateral disc swelling. PMID- 10845032 TI - Endogenous Pseudomonas endophthalmitis in an immunocompetent patient: a case for early diagnosis and treatment. PMID- 10845033 TI - Cortical blindness following pre-eclampsia. PMID- 10845034 TI - Trabeculectomy for central retinal artery occlusion. PMID- 10845035 TI - The use of a vaccine in recurrent corneal graft infection. PMID- 10845036 TI - The future of ORL-HNS and associated specialties series. The future of rhinology, anterior skull base and facial plastic surgery. PMID- 10845037 TI - A systematic approach to interpretation of computed tomography scans prior to surgery of middle ear cholesteatoma. AB - The foundation of mastoid surgery for cholesteatoma has traditionally been a thorough knowledge of the anatomy and familiarity with landmarks, constant alertness to detect unsuspected complications and the experience to tailor the surgery to the pathology encountered. Whilst not indispensable, computed tomography (CT) scanning is a useful adjunct whose potential predictive value is only truly appreciated by skilled interpretation. We present a guide to analysis to maximize the value of pre-operative radiology. PMID- 10845038 TI - Study of surfactant level in cases of primary atrophic rhinitis. AB - The surfactant system of the nose was examined biochemically in control cases and compared to cases of primary atrophic rhinitis. The study group included 25 cases with primary atrophic rhinitis compared to 10 normal volunteers. Biochemical analysis of the nasal aspirate in these cases revealed the presence of phospholipids constituting surfactant with phosphatidylcholine constituting 75.35 per cent of the total phospholipids. Biochemical analysis of the nasal aspirate in cases with primary atrophic rhinitis revealed a significant decrease in the total phospholipids compared to normal cases and also a significant change in the phospholipid profile. Thus significant biochemical changes in the surfactant system of the nose is an evident and early finding in cases of primary atrophic rhinitis. This suggests a possible role for surfactant deficiency in the aetiopathogenesis of cases of primary atrophic rhinitis. PMID- 10845039 TI - A prospective study of nasal disease in adult cystic fibrosis. AB - Twenty-six adult cystic fibrosis patients were studied to compare nasal disease with their laboratory correlates including skin testing, immunoglobulin and Aspergillus fumigatus precipitin levels, saccharin testing and sputum cultures. Six patients were asymptomatic and all of these had negative skin tests, normal IgE levels and negative Aspergillus fumigatus precipitins. Thirteen patients had rhinitis, 12 had positive skin-testing for common allergens, 10 elevated IgE levels and nine positive Aspergillus fumigatus precipitins. Seven patients had polyps, all had normal IgE levels and negative Aspergillus fumigatus precipitins, six had positive skin testing for common allergens. There also appeared to be a relationship between Pseudomonas spp. colonization and positive skin testing. PMID- 10845040 TI - Laryngeal granuloma: characteristics of the covering epithelium. AB - The purpose of the present study was to evaluate the biological behaviour of the marginal epithelium, that proliferates and eventually covers laryngeal granulomas, and to reveal the applicability of the recently re-introduced Ljubljana classification when reporting reactive epithelial hyperplastic lesions. A retrospective clinical and histomorphological analysis was performed on 149 laryngeal granuloma biopsies. Epithelial changes were classified according to the Ljubljana classification into normal epithelium; simple, abnormal, or atypical hyperplasia; and carcinoma in situ. Atrophic epithelium, not evaluated separately in the Ljubljana classification, was additionally assessed. Simple hyperplasia was found in 98 cases (65.8 per cent), abnormal hyperplasia in seven (4.7 per cent), atrophic epithelium in 24 (16.1 per cent), and normal squamous epithelium in 20 (13.4 per cent). Atypical hyperplasia and carcinoma in situ were not observed. The results of our study clearly showed that the proliferation of the covering epithelium mostly in the form of simple hyperplasia, is entirely reactive and therefore reversible. No epithelial hyperplastic lesions were found that were previously described to be associated with an increased risk of malignant alteration, namely atypical hyperplasia and carcinoma in situ. However, since an initial growth of an invasive malignant neoplasm might macroscopically imitate the appearance of laryngeal granuloma, a histological examination in all aetiological forms of laryngeal granulomas is required. By clearly discerning the benign nature of epithelial changes in laryngeal granulomas, the recently re evaluated and further formulated Ljubljana classification may also influence the clinical handling of patients. PMID- 10845041 TI - Superior laryngeal nerve preservation in peri-apical surgery by mobilization of the viscerovertebral angle. AB - Iatrogenic lesions of the superior laryngeal nerve (SLN) are much more common than is generally recognized. Since injury to this nerve may cause transient or even persistent changes either in quality of voice or in deglutition, an attempt should be made to localize and identify the nerve during surgery. This study included 74 patients who underwent surgical dissection near the thyroid apex in the neck. Effective prevention of SLN injury during surgery was achieved by anatomical localization of the nerve in the viscerovertebral angle and its functional identification with the nerve stimulator. Post-operative analysis consisted of subjective interview, rigid laryngoscopy, acoustic analysis, laryngeal videostroboscopy and cricothyroid electromyography. Four patients complained of post-operative voice changes; two were diagnosed as SLN injury (2.4 per cent), one as reflux laryngitis and the fourth as intubation granuloma. Surgical access to the SLN in the periapical area may be achieved through mobilization of the viscerovertebral angle. The use of a nerve stimulator during difficult situations should keep SLN injury at a minimum. PMID- 10845042 TI - Management strategy of mycobacterial cervical lymphadenitis. AB - The objectives of this study were to investigate the typical clinical presentation, diagnosis and treatment of mycobacterial cervical lymphadenitis (MCL). Medical records of 87 patients who were treated for MCL were retrospectively reviewed. Definitive diagnosis of MCL was made when a neck mass persisted for several weeks or months and one or more of the following was obtained: (1) positive mycobacterial cultures from biopsy material; (2) Positive mycobacterial staining of biopsy material; (3) Granulomatous inflammation and caseating necrosis on histopathological examination of biopsy material. Clinical findings were reviewed prior to treatment. The treatment included standard antituberculous medications followed by surgery in which either total excision or selective nodal dissection of the cervical lump was made. Follow-up results are presented. The chief complaint was a cervical mass that was localized mostly to the posterior cervical or submandibular regions. A fistula formation was encountered in 11.5 per cent. All patients recovered from MCL by combined antituberculous drug and surgical treatments. Clinical presentation of the disease and histopathological assessment are important in the diagnosis of MCL as well as in the differential diagnosis of tuberculous and nontuberculous MCL. Utilizing the combined medical and surgical treatment options, both tuberculous and non-tuberculous cervical adenitis can be treated successfully. PMID- 10845043 TI - Defects of the external auditory canal: a new reconstruction technique. AB - This report describes the clinical and radiographic findings together with surgical management of temporomandibular joint contents herniation through the tympanic plate of the external auditory canal. Two patients are reported. A review of the literature is presented, including a brief discussion of the embryological development of the external auditory canal. Excluding infection, trauma or neoplasm, the defect in the tympanic plate of the external auditory canal represents a developmental aberration with failure of the foramen of Huschke to close during development. A pre-auricular approach with insertion and fixation of an onlay polyethylene implant to prevent prolapse of the peri articular tissues into the ear canal is presented and described. PMID- 10845044 TI - Improving the endoscopic view of the hypopharynx with anterior neck traction during the trumpet manoeuvre. AB - The postcricoid subsite is difficult to visualize on flexible laryngopharyngoscopy. The view can be improved with either auto-insufflation manoeuvres or anterior neck skin traction. In this study, the view of the hypopharynx was graded whilst anterior neck skin traction was applied during the trumpet manoeuvre; the latter involves the patient blowing on his finger as if blowing up a balloon. On auto-insufflation alone, the postcricoid site was demonstrated in 22/25 (88 per cent) of cases and the upper oesophageal sphincter (UOS) in two out of 25 (eight per cent). Of the 22 cases in whom only the postcricoid site was demonstrated, subsequent neck skin traction revealed the UOS in eight. Overall, the use of auto-insufflation solely or in combination with traction resulted in UOS visualization in 40 per cent (10/25) of cases. The application of skin traction during trumpeting is easy to perform and should be used routinely. PMID- 10845045 TI - Oculomotor nerve palsy following submucosal diathermy to the inferior turbinates. AB - Submucosal diathermy to the inferior turbinates is a common surgical procedure performed to improve the nasal airway. We present the case of a previously well six-year-old boy who underwent submucosal diathermy and developed the unusual complication of a left oculomotor nerve palsy. The possible aetiology is discussed. PMID- 10845046 TI - Cavernous haemangioma of the nasal bones: an alternative management option. AB - The authors present a case of bilateral cavernous haemangiomas affecting the posterior ends of both inferior turbinates of the nose. The condition was treated by angiographically controlled embolization. Review of the literature back to 1967 has revealed no other report of embolization being used specifically for this condition. All previous treatments have involved surgery; we describe an alternative therapeutic option. PMID- 10845047 TI - Semi-invasive allergic aspergillosis of the paranasal sinuses. AB - Aspergillosis of the nose and paranasal sinuses has classically been divided into four types: allergic, non-invasive, invasive and fulminant. Recent reports have suggested that a semi-invasive form with bone destruction and erosion, but without fungal tissue invasion, may occur. We present a case of allergic non invasive aspergillosis of the paranasal sinuses with associated bone destruction extending into the orbit and anterior cranial fossa, in a non-immunocompromised patient. Surgical debridement combined with a prolonged course of oral itraconazole has resulted in long-term resolution with no evidence of recurrence of disease five years later. PMID- 10845048 TI - De novo laryngeal carcinoma in childhood. AB - Numerous factors contribute towards a late diagnosis of laryngeal malignancy in childhood. These include its rarity, the similarity of its early symptoms to those of other benign, common childhood conditions as well as the relative difficulty encountered during paediatric laryngeal examination. We believe that these cases are of sufficient interest when they occur to warrant reporting since the consequences of late diagnosis in these young patients can be serious. We present a case of an 11-year-old boy with advanced squamous cell carcinoma of the larynx (T3N0M0), who was successfully treated with primary total laryngectomy and bilateral selective neck dissections to avoid the potential additional morbidity of radical radiotherapy. PMID- 10845049 TI - Amyloidosis of Waldeyer's ring and larynx. AB - Amyloidosis of the upper aerodigestive tract is relatively rare. A case of localized amyloidosis involving all components of Waldeyer's ring with added laryngeal involvement is described. This has not been previously reported. A literature review of this conditions is presented. PMID- 10845050 TI - Late scar recurrence in mucoepidermoid carcinoma of base of tongue. AB - Recurrence in mucoepidermoid carcinoma of the salivary gland after 15 years is rare and present difficult therapeutic decisions. A rare case of mucoepidermoid cancer of the base of the tongue that recurred loco-regionally after 20 years and was managed judiciously by a planned combined modality approach is presented. PMID- 10845051 TI - Brown tumour of hyperparathyroidism in the mandible associated with atypical parathyroid adenoma. AB - The brown tumour of hyperparathyroidism is a localized bone tumour and an uncommon manifestation of hyperparathyroidism. A 27-year-old woman presented with a mandibular 8 x 10 cm solid mass diagnosed as central giant cell granuloma. Chemical blood analysis revealed increased serum calcium levels of 12.46 mg/dL and the parathyroid hormone level was 124 pg/dL. The patient underwent surgery with removal of a parathyroid mass. Histologically, this parathyroid tissue was seen to be limited by a fibrous capsule with morphological features consistent with atypical parathyroid adenoma. The mandibular tumour has receded and the patient declined further procedures. This is the first case reported of brown tumour as the primary manifestation of an atypical parathyroid adenoma, a lesion that shares some features with parathyroid carcinoma without the unequivocal properties of malignancy. PMID- 10845052 TI - Submandibular salivary duct cyst mimicking an external laryngocele. AB - The clinical and radiological differential diagnosis of cystic lesions of the submandibular region can be difficult. We report an unusual case of a submandibular salivary duct cyst mimicking an external laryngocele on presentation by appearing to expand on Valsalva manoeuvre, and where computed tomography (CT) scanning was unhelpful in reaching a diagnosis. We present the case, discuss the theories of pathogenesis, and review the literature on the differential diagnosis of cystic lesions in the submandibular region. PMID- 10845053 TI - Cranial neuropathy following curative chemotherapy and radiotherapy for carcinoma of the nasopharynx. AB - Cranial nerve damage following head and neck radiotherapy is an unusual event. Cranial neuropathy following concurrent chemotherapy and radiotherapy is unreported. The authors report a case of a 54-year-old man treated with curative chemotherapy and radiotherapy for a stage III nasopharyngeal carcinoma who developed an unilateral hypoglossal nerve palsy five years after therapy. Follow up examination and magnetic resonance imaging (MRI) show no evidence of recurrent disease. Hypoglossal nerve injury occurring after head and neck radiotherapy is an indirect effect due to progressive soft tissue fibrosis and loss of vascularity. This process develops over years leading to nerve entrapment and permanent damage. Cranial nerve palsies, including damage to the hypoglossal nerve, can develop years after therapy with no evidence of tumour recurrence. Chemotherapy and radiotherapy have improved progression-free and overall survival in advanced nasopharyngeal cancer. As more patients achieve long-term tumour control following chemotherapy and radiotherapy, we must be cognizant of potential late injury to cranial nerves. PMID- 10845054 TI - Excision of nasopharyngeal angiofibroma facilitated by intra-operative 3D-image guidance. AB - The latest 3D-image guidance systems to assist surgeons have greatly improved over earlier models. We describe the use of an optical infra-red system to assist in the removal of a juvenile nasopharyngeal angiofibroma. The specific advantages of this system in pre-operative assessment, intra-operative evaluation and excision of the angiofibroma are discussed. PMID- 10845055 TI - Desmoplastic fibroma of the temporal bone. AB - An unusual case of a desmoplastic fibroma of the temporal bone is presented. Although classified benign, this intraosseous lesion exhibits local aggressiveness and has a high potential for recurrence. This rare condition occurs predominantly in the mandible and in the long bones and is seldom seen in the calvarium. Here we add another case to the previously described eight in the skull and this is the fourth such case reported in the temporal bone. The clinical features, radiology, histopathology and the therapeutic considerations of this lesion in a 72-year-old female are discussed. In addition, a literature review of all the cases affecting the skull bones is presented. PMID- 10845056 TI - Ectopic thymic cyst in the neck. AB - Ectopic thymic tissue in the neck is rarely reported in medical literature. This paper presents the case of a young female, who presented with a soft, fluctuating mass in the left side of her neck. Surgical excision revealed an ectopic thymic cyst. Ectopic thymic tissue may be an infrequent finding, but it should be included in the differential diagnosis of neck masses, especially in children. This case report is accompanied by a short review of the relative literature. PMID- 10845057 TI - Cortical and subcortical contributions to activity-dependent plasticity in primate somatosensory cortex. AB - After manipulations of the periphery that reduce or enhance input to the somatosensory cortex, affected parts of the body representation will contract or expand, often over many millimeters. Various mechanisms, including divergence of preexisting connections, expression of latent synapses, and sprouting of new synapses, have been proposed to explain such phenomena, which probably underlie altered sensory experiences associated with limb amputation and peripheral nerve injury in humans. Putative cortical mechanisms have received the greatest emphasis but there is increasing evidence for substantial reorganization in subcortical structures, including the brainstem and thalamus, that may be of sufficient extent to account for or play a large part in representational plasticity in somatosensory cortex. Recent studies show that divergence of ascending connections is considerable and sufficient to ensure that small alterations in map topography at brainstem and thalamic levels will be amplified in the projection to the cortex. In the long term, slow, deafferentation dependent transneuronal atrophy at brainstem, thalamic, and even cortical levels are operational in promoting reorganizational changes, and the extent to which surviving connections can maintain a map is a key to understanding differences between central and peripheral deafferentation. PMID- 10845058 TI - Microtubule-based transport systems in neurons: the roles of kinesins and dyneins. AB - The large size and extreme polarization of neurons is crucial to their ability to communicate at long distances and to form the complex cellular networks of the nervous system. The size, shape, and compartmentalization of these specialized cells must be generated and supported by the cytoskeletal systems of intracellular transport. One of the major systems is the microtubule-based transport system along which kinesin and dynein motor proteins generate force and drive the traffic of many cellular components. This review describes our current understanding of the functions of kinesins and dyneins and how these motor proteins may be harnessed to generate some of the unique properties of neuronal cells. PMID- 10845059 TI - Apoptosis in neural development and disease. AB - Cell death via apoptosis is a prominent feature in mammalian neural development. Recent studies into the basic mechanism of apoptosis have revealed biochemical pathways that control and execute apoptosis in mammalian cells. Protein factors in these pathways play important roles during development in regulating the balance between neuronal life and death. Additionally, mounting evidence indicates such pathways may also be activated during several neurodegenerative diseases, resulting in improper loss of neurons. PMID- 10845060 TI - Gain of function mutants: ion channels and G protein-coupled receptors. AB - Many ion channels and receptors display striking phenotypes for gain-of-function mutations but milder phenotypes for null mutations. Gain of molecular function can have several mechanistic bases: selectivity changes, gating changes including constitutive activation and slowed inactivation, elimination of a subunit that enhances inactivation, decreased drug sensitivity, changes in regulation or trafficking of the channel, or induction of apoptosis. Decreased firing frequency can occur via increased function of K+ or Cl- channels. Channel mutants also cause gain-of-function syndromes at the cellular and circuit level; of these syndromes, the cardiac long-QT syndromes are explained in a more straightforward way than are the epilepsies. G protein-coupled receptors are also affected by activating mutations. PMID- 10845061 TI - The koniocellular pathway in primate vision. AB - A neurochemically distinct population of koniocellular (K) neurons makes up a third functional channel in primate lateral geniculate nucleus. As part of a general pattern, K neurons form robust layers through the full representation of the visual hemifield. Similar in physiology and connectivity to W cells in cat lateral geniculate nucleus, K cells form three pairs of layers in macaques. The middle pair relays input from short-wavelength cones to the cytochrome-oxidase blobs of primay visual cortex (V1), the dorsal-most pair relays low-acuity visual information to layer I of V1, and the ventral-most pair appears closely tied to the function of the superior colliculus. Throughout each K layer are neurons that innervate extrastriate cortex and that are likely to sustain some visual behaviors in the absence of V1. These data show that several pathways exist from retina to V1 that are likely to process different aspects of the visual scene along lines that may remain parallel well into V1. PMID- 10845062 TI - Emotion circuits in the brain. AB - The field of neuroscience has, after a long period of looking the other way, again embraced emotion as an important research area. Much of the progress has come from studies of fear, and especially fear conditioning. This work has pinpointed the amygdala as an important component of the system involved in the acquisition, storage, and expression of fear memory and has elucidated in detail how stimuli enter, travel through, and exit the amygdala. Some progress has also been made in understanding the cellular and molecular mechanisms that underlie fear conditioning, and recent studies have also shown that the findings from experimental animals apply to the human brain. It is important to remember why this work on emotion succeeded where past efforts failed. It focused on a psychologically well-defined aspect of emotion, avoided vague and poorly defined concepts such as "affect," "hedonic tone," or "emotional feelings," and used a simple and straightforward experimental approach. With so much research being done in this area today, it is important that the mistakes of the past not be made again. It is also time to expand from this foundation into broader aspects of mind and behavior. PMID- 10845063 TI - Dopaminergic modulation of neuronal excitability in the striatum and nucleus accumbens. AB - The striatum and its ventral extension, the nucleus accumbens, are involved in behaviors as diverse as motor planning, drug seeking, and learning. Invariably, these striatally mediated behaviors depend on intact dopaminergic innervation. However, the mechanisms by which dopamine modulates neuronal function in the striatum and nucleus accumbens have been difficult to elucidate. Recent electrophysiological studies have revealed that dopamine alters both voltage dependent conductances and synaptic transmission, resulting in state-dependent modulation of target cells. These studies make clear predictions about how dopamine, particularly via D1 receptor activation, should alter the responsiveness of striatal neurons to extrinsic excitatory synaptic activity. PMID- 10845064 TI - Glutamine repeats and neurodegeneration. AB - A growing number of neurodegenerative diseases have been found to result from the expansion of an unstable trinucleotide repeat. Over the past 6 years, researchers have focused on identifying the mechanism by which the expanded polyglutamine tract renders a protein toxic to a subset of vulnerable neurons. In this review, we summarize the clinicopathologic features of these disorders (spinobulbar muscular atrophy, Huntington disease, and the spinocerebellar ataxias, including dentatorubropallidoluysian atrophy), describe the genes involved and what is known about their products, and discuss the model systems that have lent insight into pathogenesis. The review concludes with a model for pathogenesis that illuminates the unifying features of these polyglutamine disorders. This model may prove relevant to other neurodegenerative disorders as well. PMID- 10845065 TI - Confronting complexity: strategies for understanding the microcircuitry of the retina. AB - The mammalian retina contains upward of 50 distinct functional elements, each carrying out a specific task. Such diversity is not rare in the central nervous system, but the retina is privileged because its physical location, the distinctive morphology of its neurons, the regularity of its architecture, and the accessibility of its inputs and outputs permit a unique variety of experiments. Recent strategies for confronting the retina's complexity attempt to marry genetic approaches to new kinds of anatomical and electrophysiological techniques. PMID- 10845066 TI - Adaptation in hair cells. AB - Hair cells adapt to sustained deflections of the hair bundle via Ca(2+)-dependent negative feedback on the open probability of the mechanosensitive transduction channels. A model posits that adaptation relieves the input to the transduction channels--force applied by elastic tip links between stereocilia--by repositioning the insertions of the links in the stereocilium. The tip link insertion and transduction channel are dragged by myosins moving on the stereocilium's actin core. This model accounts for many aspects of adaptation in hair cells of the frog saccule, where adaptation time constants are tens of milliseconds. Adaptation in hair cells of the turtle cochlea is much faster, possibly reflecting a more direct mechanism such as Ca2+ binding to the transduction channel. Adaptation mechanisms attenuate the transduction current at low frequencies and may be tuned to different corner frequencies according to the stimulus demands of the inner ear organ. Other sites of adaptation in the inner ear include accessory structures, voltage-dependent properties of hair cells, and afferent transmitter release. A remaining challenge is to understand how these processes work together to shape the output of the inner ear to natural stimuli. PMID- 10845067 TI - Mechanisms of visual attention in the human cortex. AB - A typical scene contains many different objects that, because of the limited processing capacity of the visual system, compete for neural representation. The competition among multiple objects in visual cortex can be biased by both bottom up sensory-driven mechanisms and top-down influences, such as selective attention. Functional brain imaging studies reveal that, both in the absence and in the presence of visual stimulation, biasing signals due to selective attention can modulate neural activity in visual cortex in several ways. Although the competition among stimuli for representation is ultimately resolved within visual cortex, the source of top-down biasing signals derives from a network of areas in frontal and parietal cortex. PMID- 10845068 TI - The emergence of modern neuroscience: some implications for neurology and psychiatry. AB - One of the most significant developments in biology in the past half century was the emergence, in the late 1950s and early 1960s, of neuroscience as a distinct discipline. We review here factors that led to the convergence into a common discipline of the traditional fields of neurophysiology, neuroanatomy, neurochemistry, and behavior, and we emphasize the seminal roles played by David McKenzie Rioch, Francis O Schmitt, and especially Stephen W Kuffler in creating neuroscience as we now know it. The application of the techniques of molecular and cellular biology to the study of the nervous system has greatly accelerated our understanding of the mechanisms involved in neuronal signaling, neural development, and the function of the major sensory and motor systems of the brain. The elucidation of the underlying causes of most neurological and psychiatric disorders has proved to be more difficult; but striking progress is now being made in determining the genetic basis of such disorders as Alzheimer's disease, amyotrophic lateral sclerosis, Parkinson's disease, and a number of ion channel and mitochondrial disorders, and a significant start has been made in identifying genetic factors in the etiology of such disorders as manic depressive illness and schizophrenia. These developments presage the emergence in the coming decades of a new nosology, certainly in neurology and perhaps also in psychiatry, based not on symptomatology but on the dysfunction of specific genes, molecules, neuronal organelles and particular neural systems. PMID- 10845069 TI - Plasticity and primary motor cortex. AB - One fundamental function of primary motor cortex (MI) is to control voluntary movements. Recent evidence suggests that this role emerges from distributed networks rather than discrete representations and that in adult mammals these networks are capable of modification. Neuronal recordings and activation patterns revealed with neuroimaging methods have shown considerable plasticity of MI representations and cell properties following pathological or traumatic changes and in relation to everyday experience, including motor-skill learning and cognitive motor actions. The intrinsic horizontal neuronal connections in MI are a strong candidate substrate for map reorganization: They interconnect large regions of MI, they show activity-dependent plasticity, and they modify in association with skill learning. These findings suggest that MI cortex is not simply a static motor control structure. It also contains a dynamic substrate that participates in motor learning and possibly in cognitive events as well. PMID- 10845070 TI - Guanylyl cyclases as a family of putative odorant receptors. AB - Mammals can discriminate among a large number (> 10,000) of unique odorants. The most highly supported explanation for this ability is that olfactory neurons express a large number of seven transmembrane receptors that are not spatially organized at the level of the olfactory epithelium, but whose axonal projections form a distinct pattern within the olfactory bulb. The odor-induced signaling pathway in olfactory neurons includes a Gs-like protein (G(olf)) that activates a specific adenylyl cyclase (type III) isoform, resulting in elevations of cyclic AMP and subsequent activation of a cyclic nucleotide-gated channel. The channel also can be regulated by cyclic GMP. Recently, an olfactory neuron-specific guanylyl cyclase was discovered in rodents, and subsequently a large family of sensory neuronal guanylyl cyclases was identified in nematodes. These guanylyl cyclases are concentrated in the plasma membrane of the dendritic cilia and contain extracellular domains that retain many of the primary sequence characteristics of guanylyl cyclases known to be receptors for various peptides. Thus, the guanylyl cyclases appear to represent a second family of odorant/pheromone receptors. PMID- 10845071 TI - Neural mechanisms of orientation selectivity in the visual cortex. AB - The origin of orientation selectivity in the responses of simple cells in cat visual cortex serves as a model problem for understanding cortical circuitry and computation. The feed-forward model posits that this selectivity arises simply from the arrangement of thalamic inputs to a simple cell. Much evidence, including a number of recent intracellular studies, supports a primary role of the thalamic inputs in determining simple cell response properties, including orientation tuning. This mechanism alone, however, cannot explain the invariance of orientation tuning to changes in stimulus contrast. Simple cells receive push pull inhibition: ON inhibition in OFF subregions and vice versa. Addition of such inhibition to the feed-forward model can account for this contrast invariance, provided the inhibition is sufficiently strong. The predictions of "normalization" and "feedback" models are reviewed and compared with the predictions of this modified feed-forward model and with experimental results. The modified feed-forward and the feedback models ascribe fundamentally different functions to cortical processing. PMID- 10845072 TI - Neuronal coding of prediction errors. AB - Associative learning enables animals to anticipate the occurrence of important outcomes. Learning occurs when the actual outcome differs from the predicted outcome, resulting in a prediction error. Neurons in several brain structures appear to code prediction errors in relation to rewards, punishments, external stimuli, and behavioral reactions. In one form, dopamine neurons, norepinephrine neurons, and nucleus basalis neurons broadcast prediction errors as global reinforcement or teaching signals to large postsynaptic structures. In other cases, error signals are coded by selected neurons in the cerebellum, superior colliculus, frontal eye fields, parietal cortex, striatum, and visual system, where they influence specific subgroups of neurons. Prediction errors can be used in postsynaptic structures for the immediate selection of behavior or for synaptic changes underlying behavioral learning. The coding of prediction errors may represent a basic mode of brain function that may also contribute to the processing of sensory information and the short-term control of behavior. PMID- 10845073 TI - Modular organization of frequency integration in primary auditory cortex. AB - Two fundamental aspects of frequency analysis shape the functional organization of primary auditory cortex. For one, the decomposition of complex sounds into different frequency components is reflected in the tonotopic organization of auditory cortical fields. Second, recent findings suggest that this decomposition is carried out in parallel for a wide range of frequency resolutions by neurons with frequency receptive fields of different sizes (bandwidths). A systematic representation of the range of frequency resolution and, equivalently, spectral integration shapes the functional organization of the iso-frequency domain. Distinct subregions, or "modules," along the iso-frequency domain can be demonstrated with various measures of spectral integration, including pure-tone tuning curves, noise masking, and electrical cochlear stimulation. This modularity in the representation of spectral integration is expressed by intrinsic cortical connections. This organization has implications for our understanding of psychophysical spectral integration measures such as the critical band and general cortical coding strategies. PMID- 10845074 TI - Control of cell divisions in the nervous system: symmetry and asymmetry. AB - The diverse cell types in the nervous system are derived from neural progenitor cells. Neural progenitors can undergo symmetric divisions to expand cell population or asymmetric divisions to generate diverse cell types. Furthermore, neural progenitors must exit the cell cycle in a developmentally regulated manner to allow for terminal differentiation. The patterns of neural progenitor divisions have been characterized in vertebrates and invertebrates. During the course of nervous system development, extrinsic and intrinsic cues dictate the division patterns of neural progenitors by influencing their cell cycle behavior and cellular polarity. The identification in Drosophila of asymmetrically distributed fate determinants, adapter molecules, and polarity organizing molecules that participate in asymmetric neural progenitor divisions should provide points of entry for studying similar asymmetric divisions in vertebrates. PMID- 10845075 TI - Consciousness. AB - Until recently, most neuroscientists did not regard consciousness as a suitable topic for scientific investigation. This reluctance was based on certain philosophical mistakes, primarily the mistake of supposing that the subjectivity of consciousness made it beyond the reach of an objective science. Once we see that consciousness is a biological phenomenon like any other, then it can be investigated neurobiologically. Consciousness is entirely caused by neurobiological processes and is realized in brain structures. The essential trait of consciousness that we need to explain is unified qualitative subjectivity. Consciousness thus differs from other biological phenomena in that it has a subjective or first-person ontology, but this subjective ontology does not prevent us from having an epistemically objective science of consciousness. We need to overcome the philosophical tradition that treats the mental and the physical as two distinct metaphysical realms. Two common approaches to consciousness are those that adopt the building block model, according to which any conscious field is made of its various parts, and the unified field model, according to which we should try to explain the unified character of subjective states of consciousness. These two approaches are discussed and reasons are given for preferring the unified field theory to the building block model. Some relevant research on consciousness involves the subjects of blindsight, the split brain experiments, binocular rivalry, and gestalt switching. PMID- 10845076 TI - The relationship between neuronal survival and regeneration. AB - The ability of peripheral nervous system (PNS) but not central nervous system (CNS) neurons to regenerate their axons is a striking peculiarity of higher vertebrates. Much research has focused on the inhibitory signals produced by CNS glia that thwart regenerating axons. Less attention has been paid to the injury induced loss of trophic stimuli needed to promote the survival and regeneration of axotomized neurons. Could differences in the mechanisms that control CNS and PNS neuronal survival and growth also contribute to the disparity in regenerative capacity? Here we review recent studies concerning the nature of the signals necessary to promote neuronal survival and growth, with an emphasis on their significance to regeneration after CNS injury. PMID- 10845077 TI - Neural representation and the cortical code. AB - The principle function of the central nervous system is to represent and transform information and thereby mediate appropriate decisions and behaviors. The cerebral cortex is one of the primary seats of the internal representations maintained and used in perception, memory, decision making, motor control, and subjective experience, but the basic coding scheme by which this information is carried and transformed by neurons is not yet fully understood. This article defines and reviews how information is represented in the firing rates and temporal patterns of populations of cortical neurons, with a particular emphasis on how this information mediates behavior and experience. PMID- 10845078 TI - Synaptic plasticity and memory: an evaluation of the hypothesis. AB - Changing the strength of connections between neurons is widely assumed to be the mechanism by which memory traces are encoded and stored in the central nervous system. In its most general form, the synaptic plasticity and memory hypothesis states that "activity-dependent synaptic plasticity is induced at appropriate synapses during memory formation and is both necessary and sufficient for the information storage underlying the type of memory mediated by the brain area in which that plasticity is observed." We outline a set of criteria by which this hypothesis can be judged and describe a range of experimental strategies used to investigate it. We review both classical and newly discovered properties of synaptic plasticity and stress the importance of the neural architecture and synaptic learning rules of the network in which it is embedded. The greater part of the article focuses on types of memory mediated by the hippocampus, amygdala, and cortex. We conclude that a wealth of data supports the notion that synaptic plasticity is necessary for learning and memory, but that little data currently supports the notion of sufficiency. PMID- 10845079 TI - Molecular genetics of circadian rhythms in mammals. AB - Recent gene discovery approaches have led to a new era in our understanding of the molecular basis of circadian oscillators in animals. A conserved set of genes in Drosophila and mammals (Clock, Bmal1, Period, and Timeless) provide a molecular framework for the circadian mechanism. These genes define a transcription-translation-based negative autoregulatory feedback loop that comprises the core elements generating circadian rhythmicity. This circadian core provides a focal point for understanding how circadian rhythms arise, how environmental inputs entrain the oscillatory system, and how the circadian system regulates its outputs. The addition of molecular genetic approaches to the existing physiological understanding of the mammalian circadian system provides new opportunities for understanding this basic life process. PMID- 10845080 TI - Parallel pathways for spectral coding in primate retina. AB - The primate retina is an exciting focus in neuroscience, where recent data from molecular genetics, adaptive optics, anatomy, and physiology, together with measures of human visual performance, are converging to provide new insights into the retinal origins of color vision. Trichromatic color vision begins when the image is sampled by short- (S), middle- (M) and long- (L) wavelength-sensitive cone photoreceptors. Diverse retinal cell types combine the cone signals to create separate luminance, red-green, and blue-yellow pathways. Each pathway is associated with distinctive retinal architectures. Thus a blue-yellow pathway originates in a bistratified ganglion cell type and associated interneurons that combine excitation from S cones and inhibition from L and M cones. By contrast, a red-green pathway, in which signals from L and M cones are opposed, is associated with the specialized anatomy of the primate fovea, in which the "midget" ganglion cells receive dominant excitatory input from a single L or M cone. PMID- 10845081 TI - Pain genes?: natural variation and transgenic mutants. AB - Like many other complex biological phenomena, pain is starting to be studied at the level of the gene. Advances in molecular biological technology have allowed the cloning, mapping, and sequencing of genes, and also the ability to disrupt their function entirely (i.e. via transgenic knockouts). With these new tools at hand, pain researchers have begun in earnest the task of defining (a) which of the 70,000-150,000 mammalian genes are involved in the mediation of pain, and (b) which of the pain-relevant genes are polymorphic, contributing to both natural variation in responses and pathology. Although there are only a few known examples in which single gene mutations in humans are associated with pain conditions (e.g. an inherited form of migraine and congenital insensitivity to pain), it is likely that others will be identified. Concurrently, a variety of genes have been implicated in both the transmission and control of "pain" messages in animals. The present review summarizes current progress to these ends, focusing on both transgenic (gene-->behavior) and classical genetic (behavior-->gene) approaches in both humans and laboratory mice. PMID- 10845082 TI - In vivo veritas: probing brain function through the use of quantitative in vivo biochemical techniques. PMID- 10845083 TI - Ventricular fibrillation: mechanisms of initiation and maintenance. AB - Ventricular fibrillation (VF) is the major immediate cause of sudden cardiac death. Traditionally, VF has been defined as turbulent cardiac electrical activity, which implies a large amount of irregularity in the electrical waves that underlie ventricular excitation. During VF, the heart rate is too high (> 550 excitations/minute) to allow adequate pumping of blood. In the electrocardiogram (ECG), ventricular complexes that are ever-changing in frequency, contour, and amplitude characterize VF. This article reviews prevailing theories for the initiation and maintenance of VF, as well as its spatio-temporal organization. Particular attention is given to recent experiments and computer simulations suggesting that VF may be explained in terms of highly periodic three-dimensional rotors that activate the ventricles at exceedingly high frequency. Such rotors may show at least two different behaviors: (a) At one extreme, they may drift throughout the heart at high speeds producing beat-to beat changes in the activation sequence. (b) At the other extreme, rotors may be relatively stationary, activating the ventricles at such high frequencies that the wave fronts emanating from them breakup at varying distances, resulting in complex spatio-temporal patterns of fibrillatory conduction. In either case, the recorded ECG patterns are indistinguishable from VF. The data discussed have paved the way for a better understanding of the mechanisms of VF in the normal, as well as the diseased, human heart. PMID- 10845084 TI - Basic mechanisms of atrial fibrillation--very new insights into very old ideas. AB - Atrial fibrillation (AF) was recognized and studied extensively in the early twentieth century, but many fundamental aspects of the arrhythmia were poorly understood until quite recently. It is now recognized that AF can be initiated by a variety of mechanisms that share the ability to cause extremely rapid, irregular atrial electrical activity. Once initiated, AF causes alterations in atrial electrical properties (electrical remodeling), including both rapid functional changes and slower alterations in ion channel gene expression, which promote the maintenance of AF and facilitate reinitiation of the arrhythmia should it terminate. Electrical remodeling decreases the atrial refractory period in a heterogeneous way, thus decreasing the size and stability of potential functional atrial reentry waves and promoting multiple-circuit reentry. Whatever the initial cause of AF, electrical remodeling is likely to be a final common pathway that ultimately supervenes. Recent advances in understanding ion channel function, regulation, and remodeling at the molecular level have allowed for a much more detailed appreciation of the basic determinants of AF. Improvements in the clinical management of AF will inevitably follow the recent advances in our understanding of its detailed pathophysiology. PMID- 10845085 TI - Ischemic preconditioning: from adenosine receptor to KATP channel. AB - Ischemic preconditioning is a phenomenon whereby exposure of the myocardium to a brief episode of ischemia and reperfusion markedly reduces tissue necrosis induced by a subsequent prolonged ischemia. It is hoped that elucidation of the mechanism for preconditioning will yield therapeutic strategies capable of reducing myocardial infarction. In the rabbit, the brief period of preconditioning ischemia and reperfusion releases adenosine, bradykinin, opioids, and oxygen radicals. The combined effect of the release of these substances on G proteins and the cell's phospholipases induces the translocation and activation of the epsilon isozyme of protein kinase C. Protein kinase C appears to be the first element of a complex kinase cascade that is activated during the prolonged ischemia in preconditioned hearts. Current evidence indicates that this cascade contains at least one tyrosine kinase and ultimately leads to the activation of p38 mitogen-activated protein kinase. p38 Mitogen-activated protein kinase phosphorylates mitogen-activated protein kinase-activated protein kinase 2. Mitogen-activated protein kinase-activated protein kinase 2 phosphorylates HSP27, a 27-kDa heat shock protein that controls actin filament polymerization, and, therefore, affects the integrity of the cytoskeleton. Finally, mitochondrial adenosine 5'-triphosphate-sensitive K+ channels open, and the latter may be the final mediator of protection for ischemic preconditioning. The protective pathway has many built-in redundancies, perhaps creating a safety factor. These redundancies may also explain some of the species-related differences seen in ischemic preconditioning in which one redundant pathway may predominate over another. PMID- 10845086 TI - Sodium-calcium exchange: a molecular perspective. AB - Plasma membrane Na(+)-Ca2+ exchange is an essential component of Ca2+ signaling pathways in several tissues. Activity is especially high in the heart where the exchanger is an important regulator of contractility. An expanding exchanger superfamily includes three mammalian Na(+)-Ca2+ exchanger genes and a number of alternative splicing products. New information indicates that the exchanger protein has nine transmembrane segments. The exchanger, which transports Na+ and Ca2+, is also regulated by these substrates. Some molecular information is available on regulation by Na+ and Ca2+ and by PIP2 and phosphorylation. Altered expression of the exchanger in pathophysiological states may contribute to various cardiac phenotypes. Use of transgenic approaches is beginning to improve our knowledge of exchanger function. PMID- 10845087 TI - The evolutionary physiology of animal flight: paleobiological and present perspectives. AB - Recent geophysical analyses suggest the presence of a late Paleozoic oxygen pulse beginning in the late Devonian and continuing through to the late Carboniferous. During this period, plant terrestrialization and global carbon deposition resulted in a dramatic increase in atmospheric oxygen levels, ultimately yielding concentrations potentially as high as 35% relative to the contemporary value of 21%. Such hyperoxia of the late Paleozoic atmosphere may have physiologically facilitated the initial evolution of insect flight metabolism. Widespread gigantism in late Paleozoic insects and other arthropods is also consistent with enhanced oxygen flux within diffusion-limited tracheal systems. Because total atmospheric pressure increases with increased oxygen partial pressure, concurrently hyperdense conditions would have augmented aerodynamic force production in early forms of flying insects. By the late Permian, evolution of decompositional microbial and fungal communities, together with disequilibrium in rates of carbon deposition, gradually reduced oxygen concentrations to values possibly as low as 15%. The disappearance of giant insects by the end of the Permian is consistent with extinction of these taxa for reasons of asphyxiation on a geological time scale. As with winged insects, the multiple historical origins of vertebrate flight in the late Jurassic and Cretaceous correlate temporally with periods of elevated atmospheric oxygen. Much discussion of flight performance in Archaeopteryx assumes a contemporary atmospheric composition. Elevated oxygen levels in the mid- to late Mesozoic would, however, have facilitated aerodynamic force production and enhanced muscle power output for ancestral birds, as well as for precursors to bats and pterosaurs. PMID- 10845088 TI - Variability in the size, composition, and function of insect flight muscles. AB - In order to fly, insects require flight muscles that constitute at least 12 to 16% of their total mass, and flight performance increases as this percentage increases. However, flight muscles are energetically and materially expensive to build and maintain, and investment in flight muscles constrains other aspects of function, particularly female fecundity. This review examines ways in which insects vary the size of their flight muscles, and how variation in the relative size and composition of flight muscles affects flight performance. Sources of variability in flight muscle size and composition include genetic differences within and between species, individual phenotypic responses to environmental stimuli, and maturational changes that occur before and during the adult stage. Insects have evolved a wide variety of ways to adjust flight muscle size and contractile performance in order to meet demands imposed by variation in life history and ecology. PMID- 10845089 TI - Flight respiration and energetics. AB - We use a comparative approach to examine some of the physiological traits that make flight possible. Comparisons of related fliers and runners suggest that fliers generally have higher aerobic metabolic capacities than runners but that the difference is highly dependent on the taxa studied. The high metabolic rates of fliers relative to runners, especially in insects, are correlated with high locomotory muscle cycle frequencies and low efficiencies of conversion of metabolic power to mechanical power. We examine some factors that produce variation in flight respiration and energetics. Air temperature strongly affects the flight metabolic rate of some insects and birds. Flight speed interacts with flier mass, so that small fliers tend to exhibit a J-shaped power curve and larger fliers a U-shaped power curve. As body size increases, mass-specific aerobic flight metabolism decreases in most studies, but mass-specific power output is constant or increases, leading to an increase in efficiency with size. Intraspecific studies have revealed specific genetically based effects on flight metabolism and power output and multiple ecological correlates of flight capabilities. PMID- 10845090 TI - Mechanisms underlying the cost of living in animals. AB - The cost of living can be measured as an animal's metabolic rate. Basal metabolic rate (BMR) is factorially related to other metabolic rates. Analysis of BMR variation suggests that metabolism is a series of linked processes varying in unison. Membrane processes, such as maintenance of ion gradients, are important costs and components of BMR. Membrane bilayers in metabolically active systems are more polyunsaturated and less monounsaturated than metabolically less-active systems. Such polyunsaturated membranes have been proposed to result in an increased molecular activity of membrane proteins, and in this manner the amount of membrane and its composition can act as a pacemaker for metabolism. The potential importance of membrane acyl composition in metabolic depression, hormonal control of metabolism, the evolution of endothermy, as well as its implications for lifespan and human health, are briefly discussed. PMID- 10845091 TI - Functional consequences of altering myocardial adrenergic receptor signaling. AB - From the ability to successfully manipulate the mouse genome has come important transgenic and gene-targeted knockout models that impact many areas of biomedical research. Genetically engineered mouse models geared toward the study of cardiovascular regulation have recently been described and provide powerful tools to study normal and compromised cardiac physiology. The genetic manipulation of the adrenergic receptor (AR) signaling system in the heart, including its regulation by desensitizing kinases, has shed light on the role of this signaling pathway in the regulation of cardiac contractility. One major finding, supported by several mouse models, is that in vivo contractility can be enhanced via alteration of myocardial AR signaling. Thus genetic manipulation of this critical receptor system in the heart represents a novel therapeutic approach for improving function of the failing heart. PMID- 10845092 TI - Remodeling the cardiac sarcomere using transgenesis. AB - An underpinning of basic physiology and clinical medicine is that specific protein complements underlie cell and organ function. In the heart, contractile protein changes correlating with functional alterations occur during both normal development and the development of numerous pathologies. What has been lacking for the majority of these observations is an extension of correlation to causative proof. More specifically, different congenital heart diseases are characterized by shifts in the motor proteins, and the genetic etiologies of a number of different dilated and hypertrophic cardiomyopathies have been established as residing at loci encoding the contractile proteins. To establish cause, or to understand development of the pathophysiology over an animal's life span, it is necessary to direct the heart to synthesize, in the absence of other pleiotropic changes, the candidate protein. Subsequently one can determine whether or how the protein's presence causes the effects either directly or indirectly. By affecting the heart's protein complement in a defined manner, the potential to establish the function of different proteins and protein isoforms exists. Transgenesis provides a means of stably modifying the mammalian genome. By directing expression of engineered proteins to the heart, cardiac contractile protein profiles can be effectively remodeled and the resultant animal used to study the consequences of a single, genetic manipulation at the molecular, biochemical, cytological, and physiological levels. PMID- 10845093 TI - Genetic dissection of cardiac growth control pathways. AB - Cardiac muscle cells exhibit two related but distinct modes of growth that are highly regulated during development and disease. Cardiac myocytes rapidly proliferate during fetal life but exit the cell cycle irreversibly soon after birth, following which the predominant form of growth shifts from hyperplastic to hypertrophic. Much research has focused on identifying the candidate mitogens, hypertrophic agonists, and signaling pathways that mediate these processes in isolated cells. What drives the proliferative growth of embryonic myocardium in vivo and the mechanisms by which adult cardiac myocytes hypertrophy in vivo are less clear. Efforts to answer these questions have benefited from rapid progress made in techniques to manipulate the murine genome. Complementary technologies for gain- and loss-of-function now permit a mutational analysis of these growth control pathways in vivo in the intact heart. These studies have confirmed the importance of suspected pathways, have implicated unexpected pathways as well, and have led to new paradigms for the control of cardiac growth. PMID- 10845094 TI - Genetically engineered models with alterations in cardiac membrane calcium handling proteins. AB - Regulation of intracellular Ca2+ provides a means by which the strength and duration of cardiac muscle contraction is altered on a beat-to-beat basis. Ca2+ homeostasis is maintained by proteins of the outer cell membrane or sarcolemma and the sarcoplasmic reticulum, which is the major intracellular Ca2+ storage organelle. Recently, genetic engineering techniques designed to induce specific mutations, manipulate expression levels, or change a particular isoform of various membrane Ca(2+)-handling proteins have provided novel approaches in elucidating the physiological role of these gene products in the mammalian heart. This review summarizes findings in murine genetic models with alterations in the expression levels of the sarcolemmal Ca(2+)-ATPase and Na+/Ca2+ exchanger, which move Ca2+ across the cell membrane, and the sarcoplasmic reticulum proteins, which are involved in Ca2+ sequestration (Ca(2+)-ATPase and its regulator, phospholamban), Ca2+ storage (calsequestrin), and Ca2+ release (ryanodine receptor, FK506-binding protein and junctin) during excitation-contraction coupling. Advances in genetic technology, coupled with the development of miniaturized technology to assess cardiac function at multiple levels in the mouse, have added a wealth of new information to our understanding of the functional role of each of these membrane Ca(2+)-handling proteins in cardiac physiology and pathophysiology. Furthermore, these genetic models have provided valuable insights into the compensatory cross-talk mechanisms between the major membrane Ca(2+)-handling proteins in the mammalian heart. PMID- 10845095 TI - Diapause. AB - Embryonic diapause, or delayed implantation as it is sometimes known, is said to occur when the conceptus enters a state of suspended animation at the blastocyst stage of development. Blastocysts may either cease cell division so that their size and cell numbers remain constant, or undergo a period of very slow growth with minimal cell division and expansion. Diapause has independently evolved on many occasions. There are almost 100 mammals in seven different mammalian orders that undergo diapause. In some groups, such as rodents, kangaroos, and mustelids, it is widespread, whereas others such as the Artiodactyla have only a single representative (the roe deer). In each family the characteristics of diapause differ, and the specific controls vary widely from lactational to seasonal, from estrogen to progesterone, or from photoperiod to nutritional. Prolactin is a key hormone controlling the endocrine milieu of diapause in many species, but paradoxically it may act either to stimulate or inhibit growth and activity of the corpus luteum. Whatever the species-specific mechanisms, the ecological result of diapause is one of synchronization: It effectively lengthens the active gestation period, which allows mating to occur and young to be born at times of the year optimal for that species. PMID- 10845096 TI - Multiple endocrine neoplasias. AB - The multiple endocrine neoplasia syndromes form a distinct group of genetic tumor syndromes. They include multiple endocrine neoplasia types 1 and 2, von Hippel Lindau syndrome, neurofibromatosis, and Carney complex. Research over the past decade has identified a molecular basis for each of these syndromes. This knowledge has revolutionized not only the clinical management but also has illuminated the field of human cancer research by the identification of new and important genes critical for regulation of cell growth, differentiation, and death. This review focuses on the structure, physiologic function, and molecular abnormalities of the genes involved in these syndromes. PMID- 10845097 TI - Leptin. AB - The discovery of the adipose-derived hormone leptin has generated enormous interest in the interaction between peripheral signals and brain targets involved in the regulation of feeding and energy balance. Plasma leptin levels correlate with fat stores and respond to changes in energy balance. It was initially proposed that leptin serves a primary role as an anti-obesity hormone, but this role is commonly thwarted by leptin resistance. Leptin also serves as a mediator of the adaptation to fasting, and this role may be the primary function for which the molecule evolved. There is increasing evidence that leptin has systemic effects apart from those related to energy homeostasis, including regulation of neuroendocrine and immune function and a role in development. PMID- 10845098 TI - The mechanism of action of thyroid hormones. AB - Thyroid hormone is essential for normal development, differentiation, and metabolic balance. Thyroid hormone action is mediated by multiple thyroid hormone receptor isoforms derived from two distinct genes. The thyroid hormone receptors belong to a nuclear receptor superfamily that also includes receptors for other small lipophilic hormones. Thyroid hormone receptors function by binding to specific thyroid hormone-responsive sequences in promoters of target genes and by regulating transcription. Thyroid hormone receptors often form heterodimers with retinoid X receptors. Heterodimerization is regulated through distinct mechanisms that together determine the specificity and flexibility of the sequence recognition. Amino-terminal regions appear to modulate thyroid hormone receptor function in an isoform-dependent manner. Unliganded thyroid hormone receptor represses transcription through recruitment of a corepressor complex, which also includes Sin3A and histone deacetylase. Ligand binding alters the conformation of the thyroid hormone receptor in such a way as to release the corepressor complex and recruit a coactivator complex that includes multiple histone acetyltransferases, including a steroid receptor family coactivator, p300/CREB binding protein-associated factor (PCAF), and CREB binding protein (CBP). The existence of histone-modifying activities in the transcriptional regulatory complexes indicates an important role of chromatin structure. Stoichiometric, structural, and sequence-specific rules for coregulator interaction are beginning to be understood, as are aspects of the tissue specificity of hormone action. Moreover, knockout studies suggest that the products of two thyroid hormone receptor genes mediate distinct functions in vivo. The increased understanding of the structure and function of thyroid hormone receptors and their interacting proteins has markedly clarified the molecular mechanisms of thyroid hormone action. PMID- 10845099 TI - Role of CFTR in the colon. AB - In contrast to the airways, the defects in colonic function in cystic fibrosis (CF) patients are closely related to the defect in CFTR. The gastrointestinal phenotype of CF transgenic mice closely resembles the phenotype in CF patients, which clearly indicates the crucial role of CFTR in colonic Cl- secretion and the absence of an effective compensation. In the colon, stimulation of CFTR Cl- channels involves cAMP- or cGMP-dependent phosphorylation. Exocytosis is not involved. Activation of CFTR leads to coactivation of basolateral KVLQT1-type K+ channels and inhibition of luminal Na+ channels (ENaC). In contrast to cultured cells, Ca2+ does not activate luminal Cl- channels in intact enterocytes. It activates basolateral SK4-type K+ channels and luminal K+ channels, which provide additional driving force for Cl- exit. The magnitude of Cl- secretion, however, completely depends on the presence of at least a residual CFTR function in the luminal membrane. These findings have been clearly demonstrated by Ussing chamber experiments in colon epithelium biopsies of CF and normal individuals: Colonic Cl secretion in CF patients is variable and reflects the genotype; a complete defect of CFTR is paralleled by the absence of Cl- secretion and unmasks Ca(2+) regulated K+ channels in the luminal membrane; overabsorption of Na+ in CF reflects the absence of ENaC inhibition by CFTR; and the functional status of CF colon can be mimicked by the complete suppression of cAMP stimulation in enterocytes of healthy individuals. PMID- 10845100 TI - Intracellular Ca2+ and Cl- channel activation in secretory cells. AB - Molecular and functional evidence indicates that a variety of Ca(2+)-dependent chloride (Cl(Ca)) channels are involved in fluid secretion from secretory epithelial cells in different tissues and species. Most Cl(Ca) channels so far characterized have an I- permeability greater than Cl-, and most are sensitive to 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS). Whole-cell Cl(Ca) currents show outward rectification. Single-channel current voltage relationships are linear with conductances ranging from 2 to 30 pS. Some Cl(Ca) channels are blocked by Ca(2+)-calmodulin-dependent protein kinase (CAMKII) inhibitors. Others, such as the Cl(Ca) channels of parotid and submandibular acinar cells, appear to be directly regulated by Ca2+. In native cells, the Cl(Ca) channels are located on the apical plasma membrane and activated by localized mechanisms of Ca2+ release. This positioning allows the Cl(Ca) channel to respond specifically to localized Ca2+ signals that do not invade other regions of the cell. The Cl(Ca) follows the rising phase of the Ca2+ signal, but in the falling phase hysteresis occurs where the Cl(Ca) current decays more rapidly than the underlying Ca2+. The future elucidation of the identity and mechanisms of regulation of Cl(Ca) channels will be critical to our understanding of stimulus secretion coupling. PMID- 10845101 TI - The Na-K-Cl cotransporter of secretory epithelia. AB - The Na-K-Cl cotransporters are a class of ion transport proteins that transport Na, K, and Cl ions into and out of cells in an electrically neutral manner, in most cases with a stoichiometry of 1Na:1K:2Cl. To date, two Na-K-Cl cotransporter isoforms have been identified: NKCC1, which is present in a wide variety of secretory epithelia and non-epithelial cells; and NKCC2, which is present exclusively in the kidney, in the epithelial cells of the thick ascending limb of Henle's loop and of the macula densa. Both NKCC isoforms represent part of a diverse family of cation-chloride cotransport proteins that share a common predicted membrane topology; this family also includes Na-Cl cotransporters and multiple K-Cl cotransporter isoforms. In secretory epithelia, the regulation of NKCC1, which is typically present on the basolateral membrane, is tightly coordinated with that of other transporters, including apical Cl channels, to maintain cell volume and integrity during active salt and fluid secretion. Changes in intracellular [Cl] ([Cl]i) appear to be involved in this regulation of NKCC1, which is directly phosphorylated by an unknown protein kinase in response to various secretagogues as well as reductions in [Cl]i and cell volume. This review focuses on structure-function relationships within NKCC1 and on recent developments pertaining to NKCC1 regulation at cellular and molecular levels. PMID- 10845102 TI - Chloride secretion by the intestinal epithelium: molecular basis and regulatory aspects. AB - Chloride secretion is the major determinant of mucosal hydration throughout the gastrointestinal tract, and chloride transport is also pivotal in the regulation of fluid secretion by organs that drain into the intestine. Moreover, there are pathological consequences if chloride secretion is either reduced or increased such as in cystic fibrosis and secretory diarrhea, respectively. With the molecular cloning of many of the proteins and regulatory factors that make up the chloride secretory mechanism, there have been significant advances in our understanding of this process at the cellular level. Similarly, emerging data have clarified the intercellular relationships that govern the extent of chloride secretion. The goal of our article is to review this area of investigation, with an emphasis on recent developments and their implications for the physiology and pathophysiology of chloride transport. PMID- 10845103 TI - Structure and regulation of amiloride-sensitive sodium channels. AB - Amiloride-sensitive Na+ channels constitute a new class of proteins known as the ENaC-Deg family of ion channels. All members in this family share a common protein structure but differ in their ion selectivity, their affinity for the blocker amiloride, and in their gating mechanisms. These channels are expressed in many tissues of invertebrate and vertebrate organisms where they serve diverse functions varying from Na+ absorption across epithelia to being the receptors for neurotransmitters in the nervous system. Here, we review progress made during the last years in the characterization, regulation, and cloning of new amiloride sensitive Na+ channels. PMID- 10845104 TI - Branching morphogenesis during kidney development. AB - Epithelial tissues such as kidney, lung, and breast arise through branching morphogenesis of a pre-existing epithelial structure. They share common morphological stages and a need for regulation of a similar set of developmental decisions--where to start; when, where, and in which direction to branch; and how many times to branch--decisions requiring regulation of cell proliferation, apoptosis, invasiveness, and cell motility. It is likely that similar molecular mechanisms exist for the epithelial branching program. Here we focus on the development of the collecting system of the kidney, where, from recent data using embryonic organ culture, cell culture models of branching morphogenesis, and targeted gene deletion experiments, the outlines of a working model for branching morphogenesis begin to emerge. Key branching morphogenetic molecules in this model include growth factors, transcription factors, distal effector molecules (such as extracellular matrix proteins, integrins, proteinases and their inhibitors), and genes regulating apoptosis and cell proliferation. PMID- 10845105 TI - Intrarenal dopamine: a key signal in the interactive regulation of sodium metabolism. AB - The kidney regulates sodium metabolism with extraordinary precision and sensitivity. This is accomplished by an intricate interaction between signals from extrarenal and intrarenal sources and between anti-natriuretic and natriuretic factors. Dopamine, produced in renal proximal tubule cells, plays a central role in this interactive network. Natriuretic hormones that are released from extrarenal sources, such as atrial natriuretic peptide, mediate some of their effects via renal dopamine receptors. On the level of the tubules, dopamine acts by opposing the effects of anti-natriuretic factors, such as angiotensin II and alpha-adrenergic receptors. Sodium retention leads to an increase in renal dopamine tonus, and the natriuretic effects of dopamine are more prominent under this condition. Inhibition or down-regulation of dopamine receptors significantly attenuates the natriuretic response to salt loading. Renal dopamine is modulated by the supply of filtered L-DOPA and the metabolism of dopamine via catechol-O methyldopamine. The importance of dopamine as a natriuretic hormone is reflected by its capacity to inhibit the majority of renal tubule sodium transporters. Notably, the activity of Na+, K+ ATPase is inhibited in most tubule segments by dopamine. Recent studies have elucidated many of the signaling pathways for renal dopamine receptors. Novel principles for homologous and heterologous sensitization of dopamine receptors have been detected that may explain some of the interaction between dopamine and other first messengers that modulate renal tubule sodium transport. A broad understanding of the renal dopamine system has become increasingly important, since there is now strong evidence from both clinical and experimental studies that dysregulation of the renal dopamine system plays a role in many forms of multigenetic hypertension. PMID- 10845106 TI - Endothelial signal integration in vascular assembly. AB - Regulated assembly of a highly specialized interconnecting network of vascular endothelial and supportive cells is fundamental to embryonic development and organogenesis, as well as to postnatal tissue repair in metazoans. This review advances an "endotheliocentric" model that defines tasks required of endothelial cells and describes molecular controls that regulate steps in activation, assembly, and maturation of new vessels. In addition to the classical assembly mechanisms--angiogenesis and vasculogenesis--endothelial cells are also recruited into vascular structures from the circulatory system in adult animals and from resident mesenchymally derived progenitors during organogenesis of kidney and other organs. Paracrine signaling cascades regulated by hypoxia initiate a sequentially coordinated series of endothelial responses, including matrix degradation, migration, proliferation, and morphogenetic remodeling. Surface receptors on committed endothelial lineage progenitors transduce cues from extracellular-matrix-associated proteins and cell-cell contact to direct migration, matrix attachment, proliferation, targeting and cell-cell assembly, and vessel maturation. Through their capacity to spatially segregate and temporally integrate a diverse range of extracellular signals, endothelial cells determine their migratory paths, cellular partners, and life-or-death responses to local cues. PMID- 10845107 TI - Mechanisms of guanylin action via cyclic GMP in the kidney. AB - Guanylin, uroguanylin, and lymphoguanylin are small peptides that activate cell surface guanylate cyclase receptors and influence cellular function via intracellular cGMP. Guanylins activate two receptors, GC-C and OK-GC, which are expressed in intestine and/or kidney. Elevation of cGMP in the intestine elicits an increase in electrolyte and water secretion. Activation of renal receptors by uroguanylin stimulates urine flow and excretion of sodium, chloride, and potassium. Intracellular cGMP pathways for guanylins include activation of PKG-II and/or indirect stimulation of PKA-II. The result is activation of CFTR and/or C1C-2 channel proteins to enhance the electrogenic secretion of chloride and bicarbonate. Similar cellular mechanisms may be involved in the renal responses to guanylin peptides. Uroguanylin serves as an intestinal natriuretic hormone in postprandial states, thus linking the digestive and renal organ systems in a novel endocrine axis. Therefore, uroguanylin participates in the complex physiological processes underlying the saliuresis that is elicited by a salty meal. PMID- 10845108 TI - The human language faculty as an organ. AB - Developments in the study of language and cognition give increasing credibility to the view that human knowledge of natural language results from--and is made possible by--a biologically determined capacity specific both to this domain and to our species. The functional properties of this capacity develop along a regular maturational path, such that it seems more appropriate to speak of knowledge of our own language as growing rather than as being learned. That our learning of language results from a specific innate capacity rather than by general mechanisms of induction is supported by the extent to which we can be shown to know things that we could not have learned from observation of any plausible available teaching. The domain-specificity of the language faculty is supported by the many dissociations that can be observed between control of language structure and other cognitive functions. Finally, the species specificity of the human language faculty is supported by the observation that (absent severe pathology) every human child exposed in even limited ways to the triggering experience of linguistic data develops a full, rich capacity that is essentially homogeneous with that of the surrounding community. Efforts to teach human language to other species, however, have uniformly failed. These considerations make it plausible that human language arises in biologically based ways that are quite comparable to those directing other aspects of the structure of the organism. The language organ, in this sense, can be interpreted in a functional sense, and not as implying an anatomical location comparable to that of, say, the kidney. PMID- 10845109 TI - Neural adaptation in the generation of rhythmic behavior. AB - Motor systems can adapt rapidly to changes in external conditions and to switching of internal goals. They can also adapt slowly in response to training, alterations in the mechanics of the system, and any changes in the system resulting from injury. This article reviews the mechanisms underlying short- and long-term adaptation in rhythmic motor systems. The neuronal networks underlying the generation of rhythmic motor patterns (central pattern generators; CPGs) are extremely flexible. Neuromodulators, central commands, and afferent signals all influence the pattern produced by a CPG by altering the cellular and synaptic properties of individual neurons and the coupling between different populations of neurons. This flexibility allows the generation of a variety of motor patterns appropriate for the mechanical requirements of different forms of a behavior. The matching of motor output to mechanical requirements depends on the capacity of pattern-generating networks to adapt to slow changes in body mechanics and persistent errors in performance. Afferent feedback from body and limb proprioceptors likely plays an important role in driving these long-term adaptive processes. PMID- 10845110 TI - Ligand-gated ion channel interactions with cytoskeletal and signaling proteins. AB - In recent years, it has become apparent that ligand-gated ion channels (ionotropic receptors) in the neuronal plasma membrane interact via their cytoplasmic domains with a multitude of intracellular proteins. Different classes of ligand-gated channels associate with distinct sets of intracellular proteins, often through specialized scaffold proteins containing PDZ domains. These specific interactions link the receptor channel to the cortical cytoskeleton and to appropriate signal transduction pathways in the cell. Thus ionotropic receptors are components of extensive protein complexes that are likely involved in the subcellular targeting, cytoskeletal anchoring, and localized clustering of the receptors at specific sites on the neuronal surface. In addition to structural functions, receptor-associated proteins can play important roles as activity modulators or downstream effectors of ligand-gated channels. PMID- 10845111 TI - Insights from mouse models into the molecular basis of neurodegeneration. AB - Thanks largely to cloning the genes for several neurodegenerative diseases over the past decade and the existence of mouse mutants, the molecular basis of neurodegeneration is finally beginning to yield some of its secrets. We discuss what has been learned about the pathogenesis of "triplet repeat" diseases through mouse models for spinocerebellar ataxia types 1 and 3 and Huntington disease, including the roles of nuclear aggregates and protein cleavage. We also discuss the neurologic phenotypes that arise from mutations in neurotransmitter receptors (lurcher mice) and ion channels (weaver, leaner, and tottering mice), drawing parallels between ischemic cell death and the neurodegeneration that occurs in the lurcher mouse. Finally, we discuss common mechanisms of cell death and lessons learned from these mouse models that might have broader relevance to other neurologic disorders. PMID- 10845112 TI - Spatial considerations for stimulus-dependent transcription in neurons. AB - Most neurons have elaborate dendrites as well as an axon emanating from the cell body that form synaptic connections with one or many target cells, which may be located a considerable distance from the cell body. Such complex and impressive morphologies allow some types of neurons to integrate inputs from one to many thousands of pre-synaptic partners and to rapidly propagate electrical signals, often over long distances, to post-synaptic target cells. Much slower, non electrical signals also propagate from dendrites and distal axons to neuronal nuclei that influence survival, growth, and plasticity. The distances between distal dendrites and/or distal axons and cell bodies of neurons can be hundreds of microns to more than one meter. This long-range biochemical signal propagation from distal dendrites and distal axons to neuronal nuclei is entirely unique to neurons. This review is focused on excitatory neurotransmitter signaling from dendritic synapses to neuronal nuclei as well as on retrograde growth factor signaling from distal axons to neuronal nuclei. PMID- 10845113 TI - Lung development and function in preterm infants in the surfactant treatment era. AB - Mortality of infants of < 1-kg birth weight has decreased because of surfactant treatments, antenatal glucocorticoid treatments, and new ventilation strategies. However, many of these infants develop a chronic lung disease characterized by an arrest of lung development and interference with alveolarization. Antenatal glucocorticoids can induce early lung maturation clinically, but new information from transgenic and other experimental models indicates that traditional explanations for glucocorticoid effects on the developing lung are inadequate. These very preterm infants have lungs with small lung gas volumes and delicate lung tissue that are susceptible to injury with the initiation of ventilation and subsequent ventilation. Antenatal proinflammatory exposures are frequent in very preterm infants, and postnatal injury is associated with elevations of proinflammatory cytokines in the lungs. One hypothesis is that proinflammatory cytokines can promote or interfere with lung development as well as promote lung injury. Mechanisms of lung injury being characterized in the adult lung may have unique characteristics in the developing lung. PMID- 10845114 TI - Ventilatory responses to changes in temperature in mammals and other vertebrates. AB - This article reviews the relationship between pulmonary ventilation (VE) and metabolic rate (oxygen consumption) during changes in ambient temperature. The main focus is on mammals, although for comparative purposes the VE responses of ectothermic vertebrates are also discussed. First, the effects of temperature on pulmonary mechanics, chemoreceptors, and airway receptors are summarized. Then we review the main VE responses to cold and warm stimuli and their interaction with exercise, hypoxia, or hypercapnia. In these cases, mammals attempt to maintain both oxygenation and body temperature, although conflicts can arise because of the respiratory heat loss associated with the increase in ventilation. Finally, we consider the VE responses of mammals when body temperature changes, as during torpor, fever, sleep, and hypothermia. In ectotherms, during changes in temperature, VE control becomes part of a general strategy to maintain constant relative alkalinity and ensure a constancy of pH-dependent protein functions (alphastat regulation). In mammals on the other hand, VE control is aimed to balance metabolic needs with homeothermy. Therefore, alphastat regulation in mammals seems to have a low priority, and it may be adopted only in exceptional cases. PMID- 10845115 TI - Role of transcription factors in fetal lung development and surfactant protein gene expression. AB - Branching morphogenesis of the lung and differentiation of specialized cell populations is dependent upon reciprocal interactions between epithelial cells derived from endoderm of embryonic foregut and surrounding mesenchymal cells. These interactions are mediated by elaboration and concerted actions of a variety of growth and differentiation factors binding to specific receptors. Such factors include members of the fibroblast growth factor family, sonic hedgehog, members of the transforming growth factor-beta family, epidermal growth factor, and members of the platelet-derived growth factor family. Hormones that increase cyclic AMP formation, glucocorticoids, and retinoids also play important roles in branching morphogenesis, alveolar development, and cellular differentiation. Expression of the genes encoding these morphogens and their receptors is controlled by a variety of transcription factors that also are highly regulated. Several of these transcription factors serve dual roles as regulators of genes involved in early lung development and in specialized functions of differentiated cells. Targeted null mutations of genes encoding many of these morphogens and transcription factors have provided important insight into their function during lung development. In this chapter, the cellular and molecular mechanisms that control lung development are considered, as well as those that regulate expression of the genes encoding the surfactant proteins. PMID- 10845116 TI - A century of thinking about cell membranes. PMID- 10845117 TI - Unity in diversity: a perspective on the methods, contributions, and future of comparative physiology. PMID- 10845118 TI - One-hundred years of inquiry: the mechanism of glucose absorption in the intestine. PMID- 10845119 TI - Endocrinology: survival as a discipline in the 21st century? PMID- 10845120 TI - The 20th century in respiratory physiology: one view. PMID- 10845121 TI - Toward the new millennium. PMID- 10845122 TI - Commentary on the special topic section on the use of transgenic models. PMID- 10845123 TI - One example is worth a million words. PMID- 10845124 TI - Male gender role and its implications for family medicine. PMID- 10845125 TI - There are other options. PMID- 10845126 TI - Anecdotal evidence "impressive". PMID- 10845127 TI - Family physicians and maternity care. PMID- 10845128 TI - Performing a cesarean section in the dark! PMID- 10845130 TI - Prying questions. PMID- 10845129 TI - Can we use metronidazole during pregnancy and breastfeeding? Putting an end to the controversy. AB - QUESTION: Two of my patients have been diagnosed with giardiasis, and I would like to treat them with the drug of choice for this infection, metronidazole (Flagyl). One of them is 6 weeks pregnant and the other is breastfeeding a 2 month-old infant, and I have received conflicting information about the safety of this drug during pregnancy and breastfeeding. What should I do? ANSWER: Although metronidazole has been on the market for almost 40 years, its use remains controversial during pregnancy and breastfeeding. Recent evidence has shown, however, that this drug is not associated with adverse effects during either pregnancy or breastfeeding. PMID- 10845131 TI - How accurate are they? PMID- 10845132 TI - Practice tips. Treating sexually transmitted diseases. Cooperating with a public health office helps. PMID- 10845133 TI - Are non-surgical treatments for anal fissure effective? PMID- 10845134 TI - One hundred earaches. Family practice case series. AB - OBJECTIVE: To examine whether watchful waiting was an appropriate strategy for patients with earache, when there was no clear indication to prescribe antibiotics at the first visit. DESIGN: Case series of consecutive patients with unilateral earache. SETTING: Rural family practice clinic and walk-in centre. PARTICIPANTS: One hundred patients with unilateral earache. INTERVENTIONS: Patients who clearly needed antibiotic treatment were given it; others were advised about symptom relief and were followed up as necessary. MAIN OUTCOME MEASURES: Whether patients returned to the clinic, and whether antibiotics were subsequently prescribed. RESULTS: Two patients were prescribed antibiotics at the first visit. Of the remaining 98 people, only four returned to the clinic because of earache, and two of these were prescribed antibiotics. Thus, of 100 people with earache, four received antibiotic prescriptions. CONCLUSIONS: Most people who present to primary care physicians with earache do not need antibiotics for what appears to be a mostly self-limiting condition. Both physicians and patients should be educated about this. PMID- 10845135 TI - Workshop to implement the baby-friendly office initiative. Effect on community physicians' offices. AB - OBJECTIVE: To assess the effect of a self-appraisal questionnaire and a workshop for office staff in promoting the baby-friendly office (BFO). DESIGN: A two-times three factorial design with a delayed workshop for one of two groups: an early intervention group who attended a workshop for office staff in October 1997 (n = 23) and a late-intervention group who attended in April 1998 (n = 23). Self appraisals were completed before the workshops by all participants in October 1997, by 37 offices in April 1998, and by 34 offices in October 1998. SETTING: Offices of family physicians and primary care pediatricians in Hamilton Wentworth, Ont. PARTICIPANTS: Staff of 46 offices; 74% (34/46) completed all three assessments. MAIN OUTCOME MEASURES: Degree of change in implementing each of the "10 Steps to Baby-Friendly Office" and overall average BFO score received by each office. RESULTS: Of the 34 offices completing all assessments, none followed all 10 steps. Initial mean score was 4.4 steps (standard deviation 1.4, n = 46). The workshop intervention improved overall mean scores from 4.3 to 5.6 (P < .001, n = 37). Although office staff completed the BFO self-appraisal tool, it alone had no effect on scores. Areas of improvement were noted in providing information to patients and displaying posters to promote breastfeeding. Key steps, such as not advertising breast milk substitutes and not distributing free formula, did not change. CONCLUSION: The workshop effected a modest but positive change in breastfeeding promotion. The change was maintained at 6 and 12 months after the intervention. PMID- 10845136 TI - Experience of fibromyalgia. Qualitative study. AB - OBJECTIVE: To explore illness experiences of patients diagnosed with fibromyalgia. DESIGN: Qualitative method of in-depth interviews. SETTING: Midsize city in Ontario. PARTICIPANTS: Seven patients diagnosed with fibromyalgia. METHOD: Seven in-depth interviews were conducted to explore the illness experience of patients diagnosed with fibromyalgia. All interviews were audiotaped and transcribed verbatim. All interview transcriptions were read independently by the researchers, who then compared and combined their analysis. Final analysis involved examining all interviews collectively, thus permitting relationships between and among central themes to emerge. The analysis strategy used a phenomenologic approach and occurred concurrently rather than sequentially. MAIN FINDINGS: Themes that emerged from the interpretive analysis depict patients' journeys along a continuum from experiencing symptoms, through seeking a diagnosis, to coping with the illness. Experiencing symptoms was composed of four subcategories: pain, a precipitating event, associated symptoms, and modulating factors. Seeking a diagnosis entailed frustration and social isolation. Confirmation of diagnosis brought relief as well as anxiety about the future. After diagnosis, several steps led to creation of adaptive coping strategies, which were influenced by several factors. CONCLUSION: Findings suggest that the conventional medical model fails to address the complex experience of fibromyalgia. Adopting a patient-centred approach is important for helping patients cope with this disease. PMID- 10845137 TI - Diurnal enuresis in childhood. AB - OBJECTIVE: To review the clinical classification of childhood diurnal enuresis, to describe the evaluation process, and to discuss principles of management. QUALITY OF EVIDENCE: An extensive literature review was performed with a MEDLINE search. Articles were selected according to date of publication, clinical relevance, and availability. Recent articles, cohort studies of at least 50 patients, and randomized clinical trials were preferred. Recent editions of classic textbooks were consulted. Evaluation and management activities discussed in this article are supported by original and relevant literature. MAIN MESSAGE: Most causes of childhood diurnal enuresis can be determined by a thorough history coupled with a complete physical examination and urinalysis and culture. Supplementary investigations include ultrasonography of the kidneys and bladder to screen for neurogenic bladder and urethral obstruction. When obstruction, ectopic ureter, or bladder dysfunction is suspected, voiding cystourethrography and urodynamic studies are needed. Evaluation of neurogenic bladder includes magnetic resonance imaging of the spine. Treatment is aimed at correcting poor toilet habits, preventing or treating urinary tract infections, and using appropriate medication. CONCLUSIONS: In most instances, diurnal enuresis in childhood is a benign condition with an easily identifiable cause and an excellent prognosis with time and appropriate treatment. PMID- 10845138 TI - If I shouldn't spank, what should I do? Behavioural techniques for disciplining children. AB - OBJECTIVE: To provide family physicians with a guide for evaluating discipline problems, giving suggestions for parental guidance, and diagnosing problems when discipline guidance fails. QUALITY OF EVIDENCE: A MEDLINE and PsycINFO search from 1990 to the present produced articles reviewing research on aspects of discipline. Case-control studies, expert opinion, and position statements published by the American Academy of Pediatrics and the Canadian Paediatric Society were chosen as a basis for this article. In a special supplement in 1996, a pediatric journal reviewed the controversy of spanking as an effective disciplinary method, with comments by noted researchers and clinicians. Other authors reviewed research evaluating discipline techniques. MAIN MESSAGE: Discipline problems require evaluation of children, parents, and parent-child relationships, including assessment of child development and evaluation of parenting skills and parental stressors. Parents can learn techniques more effective than spanking. Physicians can review discipline strategies and guide parents through difficult situations. Monitoring progress is important, and immediate reassessment of the situation if techniques are failing or referral to a specialist will increase the chances of a successful intervention. CONCLUSION: Discipline problems are complex and require careful assessment. Guiding parents during these types of problems requires close follow up and reevaluation when methods do not have the expected effect. PMID- 10845139 TI - Preparing your patients to travel abroad safely. Part 3: Reducing the risk of malaria and dengue fever. AB - OBJECTIVE: To provide evidence-based recommendations for family physicians advising travelers on how to reduce their risk of malaria and dengue fever. QUALITY OF EVIDENCE: A search of MEDLINE from 1990 to November 1998 found 671 articles; randomized controlled trials and systematic reviews were sought. The Cochrane Collaboration was searched for studies relevant to family physicians; meta-analyses of impregnating bed nets with permethrin were found. Health Canada's evidence-based publications were searched; 10 recommendations based on at least one well-conducted randomized trial were found. MAIN MESSAGE: Good evidence-based advice about the efficacy of mefloquine in chloroquine-resistant areas and for pregnant women and children is available, as is advice on the effectiveness of permethrin-impregnated bed nets. CONCLUSIONS: Family physicians can use evidence-based recommendations to advise their patients on how to prevent malaria. The ways in which patients neglect malaria precautions are well-known. For prevention of both malaria and dengue fever, family physicians should counsel their patients to reduce the risk of being bitten by insects. PMID- 10845140 TI - Pain relief for newborns. PMID- 10845141 TI - Physician shortages. PMID- 10845142 TI - The challenge of caring in a capitalist world. PMID- 10845143 TI - Adequacy of home care and hospital readmission for elderly congestive heart failure patients. AB - Readmission to acute care facilities is a frequent and costly problem among older adults with congestive heart failure (CHF). The study reported in this article tested the hypothesis that adequate home care, operationalized as patient perceived adequacy of formal and informal assistance, is associated with lower readmission to acute care facilities. The study followed 253 elderly (age 65 and older) Medicare patients discharged to their homes after hospitalization for CHF, through structured telephone interviews at two, six, 10, and 14 weeks postdischarge. Study findings point to the importance of home care in reducing the high risk of readmission among elderly patients. The findings raise implications for practice, policy, and research. PMID- 10845144 TI - Husbands caring for wives with dementia: a longitudinal study of continuity and change. AB - The longitudinal study reported in this article examines how the patterns of change over time for husbands who continue to care for their spouses in the community compare with husbands who place their spouses in nursing homes. There were 74 older men enrolled in the study initially; follow-up interviews were conducted with the available husbands who continued to provide care in their own homes or who had placed their spouses in nursing homes after one year. Potential theoretical relevance of the current findings and their implications for health and mental health social workers in terms of assessment, treatment, and program development for older male caregivers is highlighted. PMID- 10845145 TI - African American grandparents raising grandchildren: a national profile of demographic and health characteristics. AB - Social workers are seeing, in health care settings, an increasing number of grandparent caregivers among their clients. A disproportionate number of these are African American. This article compares the demographic and physical and mental health characteristics of African American grandparents who are raising their grandchildren with non-caregiving African American grandparents, using the National Survey of Families and Households (1992-94). Caregivers reported significantly higher levels of limitations in four of five activities of daily living (ADL) and were almost twice as likely as their peers to report clinically relevant levels of depression. PMID- 10845146 TI - Clinical social work and family medicine: a partnership in community service. AB - Traditionally, family physicians have been called on to handle the emotional problems of their patients. Their ability to address these concerns has been inconsistent. In hospital settings, where interdisciplinary teams are common, nonmedical professionals have been able to provide counseling services to patients. But this is a problem in private medical practices. This article presents a model for collaboration between family medical and clinical social work practice and describes a holistic approach for primary care practice. PMID- 10845147 TI - The NIMBY phenomenon: community residents' concerns about housing for deinstitutionalized people. AB - This article reports the findings of a study on community opposition to group homes in Montreal, Canada. This qualitative study set out to explore the underlying dynamics of what happens when a community rejects a group home. With the use of a naturalistic paradigm, three actual incidents of community opposition were studied. Nineteen interviews were conducted with community residents, elected officials, and group home developers. Community residents did not support deinstitutionalization and social integration policies and argued against group homes. The findings of this study, never reported before in previous research, have important implications for social workers and social planners. PMID- 10845148 TI - Progressives and the 2000 elections. PMID- 10845149 TI - Another look at the NIMBY phenomenon. PMID- 10845150 TI - Creating cost-efficient initiatives in social work practice in the cardiac program of an acute care hospital. AB - All three cost-saving initiatives--the creation of a one-page application form to streamline the rehabilitation application process, the use of the resource specialist to assist with applications, and the development of an information package on cardiac rehabilitation--reflect a process whereby a creative idea, generating planning, activities, and follow-up resulted in a measurable effective change in practice. This process truly translated strategy into action (Kaplan, 1996) and is vital to the current rethinking in health care of how best to do our work (Coan, 1994). Because of this process, social workers in the cardiovascular surgical division of the cardiac program are better equipped to respond to the psychosocial needs of a growing cardiac population in a fiscally restrained environment. PMID- 10845151 TI - African American Gerontology Network. PMID- 10845152 TI - Multivariate analysis of prognostic determinants after surgery for renal cell carcinoma at Himeji National Hospital. AB - A clinico-pathological study was performed retrospectively on 62 patients who underwent surgery for renal cell carcinoma between January 1992 and October 1998 at Himeji National Hospital to clarify the prognostic determinants for survival. The median follow-up period was 32 months and the cause-specific survival rates at 1, 3 and 5 years were 86.7, 81.3, 81.3%, respectively. Of the 62 patients, 11 (17.7%) patients died of renal cell carcinoma and 2 (3.2%) patients died of unrelated causes. Of the variables related to survival, presenting symptoms, C reactive protein (CRP), erythrocyte sedimentation rate (ESR), alkaline phosphatase (ALP), tumor size, pathological tumor grade, infiltration pattern, pathological tumor stage, N classification and M classification were significant risk factors for survival by univariate analysis. However, ALP, N classification and M classification were significant for survival as determined by the step-wise procedure and M classification was the most significant factor according to Cox's proportional hazard model analysis. PMID- 10845153 TI - Compensatory renal hypertrophy and changes of renal function following nephrectomy. AB - We studied the changes in the serum creatinine level and the volume of the remaining kidney following nephrectomy using contrast-enhanced compounded tomogram (CT) scans. Twenty-five patients undergoing nephrectomy for renal cell carcinoma without obvious disease in the remaining kidney were carefully followed for a period of at least two years at our hospital. Twelve patients received follow-up CT scans each year after nephrectomy. The ellipsoid volume of the kidney was calculated by measuring the 3-dimensional size on CT scans. The mean relative volume (%) of the remaining kidney increased up to year 3 postoperatively, and the final mean relative volume at varying periods from years 2 to 7 was 120%. Kidneys that were smaller prior to nephrectomy showed a tendency to have a larger final relative volume after nephrectomy, although there was no significant correlation between the kidney volume prior to nephrectomy and at final measurement. The mean serum creatinine level was significantly increased at one year after nephrectomy, but it decreased significantly over time. Therefore, both compensatory renal hypertrophy and improved renal function seemed to be established within several years after nephrectomy. However, the improvement of serum creatinine was delayed compared with the increase of kidney volume. That is, renal plasma flow might be increased early by compensatory renal hypertrophy, followed within a few years by an increase in glomerular filtration and a decrease of serum creatinine. PMID- 10845154 TI - [Transurethral removal of the ureter by the intussusception method in the treatment of renal pelvic and ureteral tumors]. AB - Between August 1986 and December 1998, 19 patients who had renal pelvic and upper ureteral tumors were treated with nephrectomy and transurethral removal of the ureter using the intussusception method. Removal of the ureter failed in 5 patients because of excessive ablasion of the ureter or insufficient electro resection around the ipsilateral ureteral orifice. Excluding those patients, the safety of the operation and the intravesical recurrence were compared with the outcome in 12 patients undergoing partial cystectomy for similar tumors. The mean operating time was not significantly shorter with the intussusception method compared with partial cystectomy (190.4 versus 251.3 minutes), but the mean blood loss was significantly smaller (187.5 versus 460.2 ml) and the intussusception method did not require a blood transfusion. The mean term of hospitalization was 20.3 days for patients treated by the intussusception method which was significantly shorter than that for patient undergoing partial cystectomy (25.4 days). Intravesical recurrence was found in seven patients (50%) treated by the intussusception method and the 1- and 5-year recurrence-free rates were 69.2% and 30.8% respectively. There was no significant difference in the recurrence-free rates between the two surgical techniques. These results suggest that the intussusception method is superior to partial cystectomy in decreasing the operating time, blood loss and term of hospitalization. It can be an attractive option in selected cases, without increasing the risk of intravesical recurrence. PMID- 10845155 TI - [Clinical study of transurethral surgery for benign prostatic hyperplasia using a thick loop (VaporTome)]. AB - From December 1996 to June 1998, we performed transurethral surgery of the prostate using a thick loop, VaporTome developed by Circon ACMI on 23 patients with symptomatic benign prostatic hyperplasia (BPH). The mean international prostate symptom score decreased significantly, from 22.6 at baseline to 9.0, 6.5, 4.9 and 5.3 at 1, 3, 6 and 12 months after operation, respectively. The mean quality of life index also decreased significantly, from 5.5 to 1.8, 1.8, 1.4 and 1.2, respectively. The mean peak flow rate increased significantly, from 6.2 preoperatively to 15.8, 17.1, 16.6 and 17.4 ml/sec at 1, 3, 6 and 12 months after operation, respectively. The mean prostate volume decreased significantly, from 66.9 to 24.8, 17.3, 19.0 and 18.2 ml. The mean residual urine decreased significantly, from 167.9 to 11.4, 9.4, 12.8 and 20.4 ml. As for overall efficacy, the rate of excellent and good cases at 1, 3, 6 and 12 months was 80.9, 89.4, 94.7 and 85.8%, respectively. No serious complications were observed. Our clinical results suggest that transurethral surgery for BPH using VaporTome has several potential advantages including high efficacy, minimal morbidity compared with standard transurethral resection of the prostate and lower cost compared with other minimally invasive procedures. PMID- 10845156 TI - [A case of Sipple syndrome whose bilateral pheochromocytomas were resected separately after a 3-year interval]. AB - A 23-year-old woman who had undergone total thyroidectomy and parabronchial lymphadenectomy at the Department of Otolaryngology in our hospital was suspected to have a thyroid carcinoma. Histological examination revealed a medullary carcinoma of the thyroid and normal parathyroid glands. Because a computed tomographic scan after surgery revealed a tumor of the left adrenal gland, the patient was examined at our department two months after the thyroidectomy, 1994. Endocrinic examinations and 123I-MIBG scintigraphy revealed a left adrenal pheochromocytoma of Sipple syndrome. She then underwent left adrenalectomy. Histological diagnosis was pheochromocytoma of the left adrenal gland. Three years after the surgery, a right adrenal tumor was detected by computed tomography. Although results of endocrine examinations were normal, 131I-MIBG scintigraphy showed a hot spot in the right adrenal gland. Right adrenal pheochromocytoma was diagnosed 3 years after surgery for a contralateral adrenal pheochromocytoma. Although surgery was recommended, the patient consented to right adrenalectomy, 1 year later after marriage and before becoming pregnant. Histological examination of the resected specimen revealed pheochromocytoma of the right adrenal gland. PMID- 10845157 TI - [Treatment of renal cell carcinoma extending into the right atrium with extra corporeal circulation using high-grade hypothermia: a case report]. AB - A 68-year-old woman underwent surgical treatment for renal cell carcinoma associated with tumor thrombus extending into the right atrium. Although the tumor thrombus reached the level of the right atrium, there were no other apparent metastases. Combination therapy with interferon alfa plus tegafur/uracil (UFT) was attempted with the expectation of reducing the tumor thrombus, but there was no change. Successful management was achieved with right radical nephrectomy, right auriculotomy, and partial cavectomy using cardiopulmonary bypass under high-grade hypothermia. After removal of the tumor and thrombus, blood loss was 13,900 ml during the patient's recovery. She had mild heart failure for about two weeks after the operation, but recovered. She was discharged on the 40th day after the operation. Proper preparation for blood transfusion is the key point of this operation. PMID- 10845158 TI - [A case report of extensively calcified renal cell carcinoma]. AB - A 73-year-old woman visited another hospital with complaints of occasional episodes of gross hematuria. A computed tomographic (CT) scan revealed extensive calcification in the left kidney, and she was referred to our hospital for further examinations. The physical examination was unremarkable other than mild back pain on her left side. Blood chemistry was normal. The CT scan revealed a mildly enhanced tumor of 6 cm in diameter accompanied by extensive calcification in the left kidney. Angiography revealed a hypovascular tumor in the left kidney. A left radical nephrectomy was carried out. Gross appearance of the specimen showed extensive calcification from the renal pelvic to the parenchyma of the lower pole of the left kidney. The histopathological diagnosis was renal cell carcinoma, alveolar type, mixed subtype, pT2pN0pM0. PMID- 10845159 TI - [Spontaneous rupture of infected renal cyst: a case report]. AB - A 68-year-old man who had been followed at 1-year intervals for a left giant renal cyst was referred to our hospital for left flank pain and fever elevation. Abdominal computed tomographic scan revealed a giant cystic lesion of the left kidney suspected to be communicating with the urinary tract. Percutaneous puncture of this lesion was performed and the fluid was drained. The fluid was yellowish and cloudy, and E. coli was detected by its culture. Injection of contrast medium showed communication between the cyst and the urinary tract. The patient underwent drainage for a ruptured renal cyst. Rupture of a renal cyst is uncommon, and this is the 16th case reported in Japan. PMID- 10845160 TI - [A case of recurrent spontaneous vesical rupture subsequent to irradiation for uterine cancer]. AB - The patient, a 68-year-old woman, had undergone radical hysterectomy and post operative radiotherapy for uterine cancer in 1974. She was admitted to our hospital complaining of abdominal pain in February, 1996. Contrast-enhanced computed tomographic scan and cystography showed leakage of contrast medium around the bladder. We diagnosed her with spontaneous vesical rupture, and performed conservative therapy. Two months later, she was re-admitted with recurrent vesical rupture. We again performed conservative therapy. We consider that conservative therapy can be indicated for spontaneous vesical rupture with good general condition and no severe urinary tract infection. PMID- 10845161 TI - [Papillary adenocarcinoma of the prostate: report of 3 cases]. AB - We report here 3 cases of papillary adenocarcinoma of the prostate. In all 3 cases, the tumors were discernible on cystourethroscopy and transurethral biopsy established the diagnosis, whereas no significant finding was found on digital rectal examination. Although androgen deprivation therapy was administered in all cases, different surgical procedures were employed according to the stage in each case. In case 1, since the papillary tumor was confined within the prostatic urethra, complete resection was accomplished by transurethral resection (TUR). In case 2, since pelvic lymph nodes metastases were found, local radiation therapy was added. In case 3, since the patient had vesical invasion of tumor total cysto prostatectomy was performed. Papillary adenocarcinoma of the prostate originates from the prostatic duct, resulting in existence at the "central portion" of the prostate gland. Cystourethroscopy and transurethral biopsy is helpful for diagnosis of this disease, whereas rectal digital examination is useless. As a surgical procedure for the primary site, TUR may be efficient for tumors confined within the prostatic urethra, although more extensive surgery may be necessary for those with a more invasive profile. PMID- 10845162 TI - [Urethral injury with unusual clinical course: a case report]. AB - A 20-year-old men fell from a ladder in 1995, striking his perineum strongly, and macroscopic hematuria with painful urination lasted for several days. Subsequently, swelling of the perineum on urination continued to occur. However, he did not seek medical treatment until July 1997, when he consulted our medical institution. A diverticulum-like change was found in the bulbous urethra on urethrocystography, and a tear in the same position on urethroscopy. On September 22, 1997, surgery was performed after constructing a cystostomy, the tear was located using a urethroscope, and closed through a perineal approach. At 14 days after surgery, there was extra-urethral leakage of contrast media on voiding cystourethrography, and at 28 days after surgery, the findings showed improvement. Since then, he has been free of the preoperative symptoms. PMID- 10845163 TI - [Delayed posttraumatic high flow priapism: a case report]. AB - A 21-year-old male complained of persisting erection. A urethral balloon catheter had been for 3 weeks after indwelt urethral injury by a skateboard, and he was hospitalized because of penile erection persisting after removing the catheter. High flow priapism was suspected by intracavernous blood gas study and color Doppler ultrasound study. Selective internal pudendal arteriography revealed a leakage of contrast medium at the base of penis. He was treated with selective embolization of bilateral internal genital arteries using gelatin sponges and achieved detumescence. Normal potency was evident 3 months later by examining nocturnal penile tumescence. PMID- 10845164 TI - [A case of testicular tumor in a patient with Down's syndrome: a case report]. AB - A 40-year-old man with Down's syndrome was admitted to our hospital for complaint of painless, swollen right scrotum. Computed tomography revealed a right testicular tumor and mass of iliac fossa. Right high orchiectomy and extirpation of mass were performed. Histological diagnosis of right testicular tumor was seminoma and the mass was an external iliac lymph node metastasis of seminoma. After operation, radiation therapy was performed. PMID- 10845165 TI - [A case of Down's syndrome associated with testicular seminoma]. AB - Down's syndrome is an inherited disorder caused by trisomy of chromosome 21. In patients with Down's syndrome, an increased risk of leukemia has been observed. Recently, the coincidence of testicular cancer with this syndrome has been also emphasized. We present a case of Down's syndrome associated with testicular seminoma. This is the 19th case of Down's syndrome associated with testicular tumor in Japan. PMID- 10845166 TI - [A case of malignant lymphoma of the epididymis]. AB - A 17-year-old man visited our hospital with the chief complaint of painless swelling of the left scrotal content. An elastic hard mass was palpable at the upper pole of the left testis. Left radical orchiectomy was performed. The tumor originated from the epididymis and did not involve the testis or the spermatic cord. Histologically, the tumor was diagnosed as a malignant lymphoma (non Hodgkin's lymphoma, diffuse mixed cell type, B-cell type). No abnormalities were found in other organs. After establishment of the diagnosis of primary epididymal malignant lymphoma, 3 courses of chemotherapy (adriamycin, vincristine, cyclophosphamide, prednisolone) were performed. No evidence of recurrence or metastasis was found 26 months after surgery. PMID- 10845167 TI - Comprehensive gynecologic endoscopic hospital privileging program. Implementation and assessment. AB - OBJECTIVE: To describe a comprehensive gynecologic endoscopic privileging program at an urban teaching hospital and evaluate its effect on complication rates. STUDY DESIGN: In 1996, a gynecologic endoscopy privileging program was instituted. Initially, experienced surgeons were invited to apply for advanced privileges based on submission of a case list. Afterwards, new applications were approved by proctorship. Since 1995, charts have been reviewed using the following indicators; operating time, estimated blood loss, length of stay, readmission, diagnosis of cancer, reexploration and admission for hysteroscopic fluid overload. Cases were also independently identified when a major vascular or visceral injury occurred. RESULTS: Among the 3,880 gynecologic endoscopic procedures performed during the review period, 2,702 medical records were randomly screened. Following institution of the program, there was no change noted in rates of hysteroscopic fluid overload, readmission, reexploration or unrecognized diagnosis of cancer. However, a decrease was noted in excess blood loss (odds ratio [OR] 0.6, 90% confidence interval [CI] 0.4, 0.9) and operating time greater than four hours (OR 0.6, CI 0.4, 0.9). Length of hospital stay was also reduced in the year following implementation of the privileging process (OR 0.2, CI 0.1, 0.3). Fifty-four cases of visceral or major vascular injury occurred during the three-year period. The risk of visceral injury revealed a trend from 1.9% to 1.0% after institution of the privileging process (OR 0.5, CI 0.3, 1.0). CONCLUSION: Establishment of a comprehensive gynecologic endoscopic hospital privileging program was associated with a reduction in rates of excess blood loss and operating times and a decreasing trend in visceral injuries. PMID- 10845168 TI - Economic and clinical outcomes of microlaparoscopic and standard laparoscopic sterilization. A comparison. AB - OBJECTIVE: To compare micro-laparoscopic surgical sterilization and standard laparoscopic sterilization with respect to cost effectiveness and patient preferences. STUDY DESIGN: A retrospective study of all laparoscopic surgical sterilizations performed under general anesthesia at Johns Hopkins Bayview Medical Center--16 micro-laparoscopies and 34 standard laparoscopies. Cases selected for review were limited to patients undergoing surgical contraception and not requiring additional, concurrent procedures. Laparoscopic surgical sterilization was performed using a double-puncture technique with silicone band application. In each case either a standard, 10-mm laparoscope or a 2-mm micro laparoscope was used, and the procedure was performed under general anesthesia. Postoperative pain management was achieved by nonsteroidal antiinflammatory drugs and/or narcotic analgesia. All cases were performed by residents under faculty supervision. Medical records and hospital billing records were reviewed, and a standardized telephone interview was conducted to assess postoperative quality of life and patient satisfaction. RESULTS: Both techniques were comparable in cost effectiveness. There was no significant difference in operating room time, average operating room costs, average ancillary department costs, instrument and supply costs, or length of stay. Postoperative discomfort was significantly less with microlaparoscopy (P = .05), and patient satisfaction was higher in the microlaparoscopy group. CONCLUSION: Microlaparoscopy and the standard laparoscopic approach for surgical sterilization are associated with similar hospital charges. Postoperative pain and overall patient satisfaction were significantly better with microlaparoscopy than standard laparoscopy. PMID- 10845169 TI - Radiologic diagnosis and surgical treatment of urethral diverticulum in women. A reappraisal of voiding cystourethrography and positive pressure urethrography. AB - OBJECTIVE: To reappraise the optimum radiologic procedure for the diagnosis of urethral diverticula in women and to assess the results of surgery. STUDY DESIGN: From January 1994 to December 1997, 132 urine-continent women with pelvic prolapse and the most characteristic symptoms of urethral diverticulum underwent pelvic repair. In addition to routine urologic evaluations and pelvic examinations, the women underwent voiding cystourethrography (VCUG) and positive pressure urethrography (PPUG) with a double-balloon catheter. The x-ray examination was scheduled as follows. One hundred thirty-two women initially underwent VCUG followed by a PPUG. Another VCUG was subsequently performed on women with a negative PPUG finding. Twelve patients were excluded because they were unable to urinate for the first VCUG; thus, the study group was composed of the remaining 120 patients. Subsequently those patients underwent various pelvic operations, including total abdominal/vaginal hysterectomy, abdominal colposacropexy, Manchester's operation, sacrospinous ligament suspension, diverticulectomy and colporrhaphy. RESULTS: Both VCUG and PPUG were able to demonstrate the diverticula in 20 of the 120 women. Another 13 women showed positive findings on PPUG. Ten of the 87 cases with a negative PPUG had a positive finding in the subsequent VCUG. Forty-two diverticula were detected in 39 patients during pelvic repair. The only complication of urethral diverticulectomy was urethrovaginal fistula, which occurred in 3 of the 39 patients. CONCLUSION: Though VCUG is not as sensitive as PPUG, it still has merits and can be used as a screening test. If VCUG is inconclusive but clinical suspicion persists, particularly when PPUG is doubtful, magnetic resonance imaging should be considered. PMID- 10845170 TI - Sperm cryopreservation with empty human or mouse zona pellucidae. A comparison. AB - OBJECTIVE: To compare empty zona pellucidae (ZP) and human of mouse oocytes for sperm cryopreservation. STUDY DESIGN: ZP was prepared by evacuating the cytoplasm of human or mouse oocytes. The evacuated ZP were injected with spermatozoa from patients with severe oligoasthenozoospermia from normally fertile men. After a freezing-thawing procedure, the spermatozoa were aspirated outside the ZP. According to the different origins of ZP and sperm, the spermatozoa number per ZP, motile sperm number before freezing and after thawing, lost sperm number per ZP after freezing and sperm recovery rate were compared. RESULTS: Spermatozoa number, motile sperm number before freezing and after thawing, lost sperm number per ZP and sperm recovery rate in all groups were comparable. The total mean motile sperm number before freezing and after thawing, mean lost sperm number after thawing and sperm recovery rate were 15.1%, 12%, 1.6% and 80%, respectively. CONCLUSION: ZP is an ideal vehicle for cryopreservation of sperm collected from patients with severe oligoasthenozoospermia. There were no differences between human and mouse ZP for sperm storage. PMID- 10845171 TI - Postoperative adhesion prevention with an oxidized regenerated cellulose adhesion barrier in infertile women. AB - OBJECTIVE: To evaluate the efficacy of an oxidized regenerated cellulose adhesion barrier as an adjuvant in preventing postoperative adhesions in infertile women undergoing reconstructive surgery. STUDY DESIGN: Thirty-eight cases of reconstructive surgery that could be followed up for more than two years (myomectomy 19, cystectomy 5, tuboplasty 10, uteroplasty 4) at the Fujita Health University Hospital were evaluated retrospectively. The barrier (Intercede, Johnson & Johnson) was used to cover the surgical site in 23 of these cases (Intercede group); no adjuvant was used in 15 cases, which represent the surgical control group (Intercede - group), including 23 second-look operation cases (16 in the Intercede and 7 in the control group). Postoperative adhesion prevention and pregnancy rates were estimated. RESULTS: At the second-look operation, six cases (37.5%) in the Intercede + group and six (85.7%) in the Intercede - group had postoperative adhesions. No significant difference was found in either intensity or area covered with adhesions between the two groups. Eighteen cases (78.3%) in the Intercede and seven (46.7%) in the Intercede - group conceived during the follow-up period. CONCLUSION: The use of Intercede significantly reduced the rate of postoperative adhesion formation, with a statistically significant increase in the pregnancy rate as compared to the surgical controls. PMID- 10845172 TI - Clinical and sonographic fetal weight estimates in active labor with ruptured membranes. AB - OBJECTIVE: To determine and compare the accuracy of clinical and sonographic estimates of fetal weight (EFW) in active labor with ruptured membranes. STUDY DESIGN: Clinical and sonographic EFWs were obtained on 107 term patients in active labor, with cervical dilatation of 4 cm or more and ruptured membranes. Accuracy of birth weight was determined by calculating percentage error, absolute percentage error and ratio of estimates within 10% of actual birth weight for all stages of labor. Statistical analysis was by paired t test, Wilcoxon sign test, chi 2 test and Mann-Whitney U test; P < .05 was considered significant. RESULTS: Absolute percentage errors were lower by the sonographic method at all stages. Except for the second stage of labor, the rates of birth weight +/- 10% were higher with the sonographic method than with the clinical method (83.17% vs. 60.75% and 84.9% vs. 63.44%, respectively). Estimations performed in the first stage were more accurate than in the second stage with both methods (absolute error of 7.82 +/- 5.5 vs. 12.38 +/- 4.9 for clinical and 5.44 +/- 5.99 vs. 9.08 +/- 3.19 for sonographic). CONCLUSION: During active labor with ruptured membranes, sonographic EFWs are more accurate than clinical estimations. The accuracy of both methods is reduced during the second stage of labor. PMID- 10845173 TI - Cesarean delivery for suspected fetal distress among preterm parturients. AB - OBJECTIVE: Among preterm parturients (< 37 weeks) who underwent cesarean delivery for suspected fetal distress, to determine the factors associated with decision incision time (DIT) of < or = 30 minutes and to assess if umbilical arterial pH < 7.10 is more common with DIT < or = 30 or > 30 minutes. STUDY DESIGN: The peripartum course of all patients who had cesareans for suspected fetal distress over three years was reviewed. The inclusion criteria were reliable gestational age < 37 weeks and a single indication for cesarean delivery, suspected fetal distress. Twenty antepartum and intrapartum factors were used in a univariate analysis. RESULTS: The mean DIT among the 84 parturients was 30.5 +/- 21.2 minutes, and 63% of patients had surgery started within 30 minutes. The incidence of pH < 7.10 was 20%. Multivariate analysis indicated that the two factors significantly associated with prolonged time to surgery were tachycardia with decreased variability (odds ratio [OR] 5.9, 95% confidence interval [CI] 1.6 21.6) and use of spinal anesthesia (OR 6.2, 95% CI 1.1-35.0). Though none of the 20 variables had significant univariate associations with neonatal acidosis at alpha = .05, those with P < .20 were considered in multiple logistic regression analysis. None of the 20 factors were associated with pH < 7.10, including DIT of > or = 30 minutes (OR 0.26, 95% CI 0.06-1.03). CONCLUSION: DIT is likely to be > 30 minutes if cesarean delivery is due to decreased fetal heart variability or if spinal anesthesia is utilized; neonatal acidosis, however, is not significantly associated with a prolonged interval. PMID- 10845174 TI - Bioavailability of iron in two prenatal multivitamin/multimineral supplements. AB - OBJECTIVE: To determine the iron bioavailability in two popular prenatal multivitamin/multimineral supplement tablets containing 27 mg elemental iron. STUDY DESIGN: Iron absorption during an eight-hour period following ingestion of a multivitamin/multimineral formulation, both fasting and with a standardized meal, was measured in a group of 30 pregnant women (24-32 weeks of gestation) and statistically compared. The prenatal formulations were Stuartnatal Plus and Materna (Wyeth-Ayerst Pharmaceuticals, Philadelphia, Pennsylvania), and each contains 27 mg of elemental iron. A placebo was included in the study for the control group in this crossover, single-blind study. RESULTS: The net iron bioavailability (mean +/- SE) of Stuartnatal Plus and Materna, accounting for diurnal variation, and the iron ingested with the standardized meal was 5.4 +/- 0.4 and 4.6 +/- 0.2 mg, respectively, while fasting and 2.9 +/- 0.4 and 2.7 +/- 0.4 mg, respectively, postprandially. The total amount of iron absorption in the fasting states from both prenatal formulations exceeded the 3 mg of supplemental iron absorption per day recommended by the National Academy of Sciences. CONCLUSION: The results of this study indicate that these two prenatal multivitamin/multimineral formulations provide > 3.0 mg of supplemental iron absorption (fasting) as recommended by the National Academy of Sciences and 2.7 mg of iron absorption above the levels achieved following ingestion of a standard, low-iron test meal. PMID- 10845175 TI - Detection of a human parvovirus intrauterine infection with the polymerase chain reaction. AB - OBJECTIVE: To detect human parvovirus B19 intrauterine infection in pregnancy with the polymerase chain reaction (PCR). STUDY DESIGN: DNA of chorionic villi and amniotic fluid was extracted and the gene of human parvovirus B19 amplified with PCR. RESULTS: The study analyzed 61 specimens of chorionic villi and 26 specimens of amniotic fluid and found two positive specimens of chorionic villi and 1 positive specimen of amniotic fluid. CONCLUSION: The vertical transmission of human parvovirus B19 infection in early pregnancy may be a pathway of intrauterine infection. Chorionic villus sampling in early pregnancy and PCR could be developed as a method of prenatal diagnosis of human parvovirus B19 intrauterine infection. PMID- 10845176 TI - Intraoperative ultrasound guidance for operative hysteroscopy. A prospective study. AB - OBJECTIVE: To evaluate the efficacy and safety of ultrasound (US) guidance as compared to laparoscopic monitoring during operative hysteroscopy. STUDY DESIGN: Prospective, open study including 81 patients undergoing operative hysteroscopy under US guidance for uterine septum and submucous myoma. Clinical and surgical outcomes were compared with those in an historical control group of 45 patients undergoing the same operation under laparoscopic guidance. RESULTS: US guidance proved satisfactory in all patients, and there were no complications due to insufficient visualization of the pelvic structures; in no case was conversion to laparoscopic guidance required. US scanning was most useful in determining the outer limit of the intramural component of submucous partial intramural myoma, allowing complete resection. During metroplasty, US guidance allowed extension of the resection beyond the normal limit conventionally defined by hysteroscopy; none required reintervention. By comparison, in the control group, a second attempt was required because the operation was insufficiently radical in four patients. CONCLUSION: US guidance was used successfully as the only visual aid for hysteroscopic surgery, comparing favorably with laparoscopy in terms of efficacy and safety. PMID- 10845177 TI - Determining risk factors for intrapartum fetal death. AB - OBJECTIVE: To determine the antenatal and intrapartum risk factors for intrapartum fetal death (IPFD). STUDY DESIGN: We analyzed 72,875 singleton deliveries > 500 g between the years 1990 and 1997. In order to find independent factors contributing to IPFD, a multiple logistic regression model was constructed. RESULTS: During the study period there were 64 cases of IPFD. Birth weight was inversely associated with IPFD in the multivariate analysis. Other significant factors in the multivariate analysis were mothers older than 35 years, polyhydramnios, congenital malformations, pathologic presentation, abruptio placentae and cord prolapse. No association was found with maternal diabetes or hypertension. CONCLUSION: IPFD was significantly associated with low birth weight, older maternal age, polyhydramnios, congenital malformations and pathologic presentation. Direct causes were abruptio placentae and cord prolapse. Pregnancies in older women and those complicated by polyhydramnios and pathologic presentation should be carefully evaluated during pregnancy and delivery in order to decrease the incidence of IPFD. PMID- 10845178 TI - Involvement of drugs in sexual assault. AB - OBJECTIVE: To obtain information about the relationship of alcohol and drug usage in victims of sexual assault, including the newly identified "date rape" drugs gamma hydroxybutyrate and flunitrazepam. STUDY DESIGN: Analysis of urine samples with gas chromatography combined with mass spectrometry can identify alcohol and numerous other drugs with a high degree of specificity. This service was offered to rape treatment centers throughout the United States in May 1996; urine samples obtained from sexual assault victims suspected of drug use by history or physical examination were sent for testing at the discretion of the examiner. RESULTS: As of March 1999, a total of 2,003 specimens were analyzed. Nearly two-thirds of the samples contained alcohol and/or drugs; the predominant substances found were alcohol, present in 63%, and marijuana, present in 30%. A substantial subset of the specimens was found to contain other illicit substances, frequently in combination. GHB and flunitrazepam were found in < 3% of the positive samples. Additionally, over the two-year study period, the use of these two drugs appeared to be declining. CONCLUSION: These findings support prior data indicating that alcohol, marijuana and/or other drugs are important risk factors in sexual assault. Continued monitoring of drug use by victims of sex crimes is important, and programs that serve victims should modify protocols to reflect this. PMID- 10845179 TI - Posthysterectomy bleeding after ketorolac tromethamine and spinal anesthesia. A case report. AB - BACKGROUND: Ketorolac is a very useful medication in gynecologic surgery, providing pain relief and less narcotic use. This greatly shortens the recovery time and length of hospital stay. As with other nonsteroidal antiinflammatories, gynecologists are aware of the potential for bleeding problems from ketorolac, yet those bleeding complications have rarely been seen in gynecology and have not been previously reported in the gynecologic literature. A recent case demonstrates the serious complication that can occur when ketorolac inhibits platelet function postoperatively. CASE: A 36-year-old woman began bleeding intraabdominally shortly after an otherwise-unremarkable vaginal hysterectomy done under spinal anesthesia. CONCLUSION: A literature review suggests that this complication is highly associated with nongeneral anesthesia surgery, such as spinal, epidural and conscious sedation. This fact, however, has not been published in the gynecologic literature and is not well known to gynecologists. Pending further elucidation of this problem, avoidance of ketorolac perioperatively with nongeneral anesthesia cases should be considered. PMID- 10845180 TI - Incidental finding of an accessory ovary in a 16-year-old at laparoscopy. A case report. AB - BACKGROUND: Supernumerary and accessory ovaries are rare anomalies. The reported incidence of these conditions is 1:29,000-700,000 gynecologic admissions. Since 1864 there have been only six cases of accessory ovary reported in the literature. Additionally, there have been 26 reported cases of supernumerary ovaries. This case seems to be the seventh reported case of an accessory ovary in over 135 years. It is the third case in a woman under age 18. In 36% of reported cases, associated congenital anomalies have been identified. Defects have included accessory fallopian tube, bifid fallopian tubes, accessory tubal ostium, bicornuate and unicorunate uteri, septate uterus, agenesis of kidney or ureter, bladder diverticulum, accessory adrenal gland and lobulated liver. CASE: A 16 year-old nulligravida complained of persistent left lower quadrant abdominal pain. Abdominal ultrasound confirmed a left ovarian cyst, which was unresponsive to medical management with oral contraceptives. At laparoscopy, a normally placed, right ovary and eutopic left ovary were identified. In addition, an accessory ovary was identified on the posterior surface of the left broad ligament. CONCLUSION: Since accessory ovaries are likely to be asymptomatic, they may be underreported. This condition is associated with a high risk of pelvic and renal anomalies and should lead to further evaluation to allow physicians to provide advice about future reproductive function and management of congenital anomalies. PMID- 10845181 TI - Vulvar arteriovenous hemangioma. A case report. AB - BACKGROUND: Vulvar arteriovenous hemangioma is a rare pathologic lesion that can have a unique clinical presentation. One manifestation of this lesion has not previously been described in the literature. CASE: A painful vulvar mass occurred in a healthy woman, with cyclic perineal discomfort and enlargement of the mass with menses. There was no obvious lesion on the skin. Excisional biopsy and groin exploration revealed a vascular lesion in the subcutaneous layer, consistent with arteriovenous hemangioma. The mass was successfully treated by excision, and the patient was asymptomatic 12 months later. CONCLUSION: Surgical exploration should be considered when a definitive diagnosis of a symptomatic vulvar mass is uncertain. In this case, excision of the mass allowed diagnosis and definitive treatment. PMID- 10845182 TI - Immunologic evaluation of an Escherichia coli O157-infected pregnant woman. A case report. AB - BACKGROUND: To the best of our knowledge, this is the first case of pregnancy complicated by hemorrhagic enterocolitis due to Escherichia coli O157. CASE: A woman with hemorrhagic enterocolitis due to E coli O157 was seen at 32 weeks of gestation. We investigated her immune response to 0157 lipopolysaccharide and to Shiga toxin in the sera and breast milk. CONCLUSION: IgM and IgA to 0157 lipopolysaccharide in the breast milk of this patient might protect her infant after the disappearance of serum IgM. PMID- 10845183 TI - Mutagenicity of bovine lactoferrin in reverse mutation test. AB - The mutagenicity of bovine lactoferrin, which is an iron-binding glycoprotein in milk, was evaluated by the Ames mutagenicity test. A total of 5 test strains including 3 base-pair substitution-type strains, Salmonella typhimurium TA100, TA1535 and Escherichia coli WP2uvrA, and 2 frameshift-type strains, TA98 and TA1537, were used in the test. The test was performed by both the direct method and the metabolic activation method with preincubation applied in each instance. The concentration range of the test solution was 0.16 to 5.00 mg/100 microliters (plate). Results of the test revealed that the number of revertant colonies at each concentration of the test solutions was less than 1.4 times that of the control group. In the test system used, bovine lactoferrin did not exhibit mutagenicity. PMID- 10845184 TI - Large amount of vitamin A has no major effects on thyroidal hormone synthesis in two-stage rat thyroid carcinogenesis model using N-bis(2 hydroxypropyl)nitrosamine and thiourea. AB - In our previous investigation, which focused on two-stage carcinogenicity in the thyroid, rats were administered N-bis(2-hydroxypropyl)nitrosamine (DHPN), followed by thiourea (TU) over an experimental period of 19 weeks. Simultaneous treatment with a high level of vitamin A (VA) enhanced the induction of proliferative lesions that originated from the thyroidal follicular epithelium. To examine whether hormone synthesis in the thyroid could be inhibited by simultaneous treatment with a large amount of VA and TU, all of the rats were initially given a single subcutaneous injection of 2,800 mg DHPN/kg followed by a supply of 0% TU + 0% VA (DHPN only, control group), 0.2% TU in their drinking water (DHPN/TU group), 0.1% VA in their diet (DHPN/VA group), or 0.2% TU + 0.1% VA (DHPN/TU + VA group) during an experimental period of 4 weeks. Results obtained indicate that the iodine uptake and organification, namely iodination of tyrosine residue in thyroglobulin, of the thyroid, were significantly decreased in the DHPN/TU group compared to the DHPN control group. The variation in these values was attributable to the inhibitory effect of TU upon thyroid hormone synthesis. Results obtained from the DHPN/TU + VA and DHPN/TU groups were comparable. Therefore, the possibility that modification of hormone synthesis contributes to the enhancing effect of simultaneous treatment with a large amount of VA on thyroidal tumor induction by TU is considered to be very minimal. PMID- 10845186 TI - Effects of valproic acid on fertility and reproductive organs in male rats. AB - Crj:CD(SD)IGS rats were orally administered valproic acid at doses of 250, 500 or 1000 mg/kg/day for 4, 7 or 10 weeks. At each dose, one group of male rats was euthanized after 4-week dosage (4-week dose group) and the other two were mated with untreated females after 4 (7-week dose group) or 7 (10-week dose group) weeks of treatment with valproic acid and their fertility was evaluated. Females were euthanized on day 14-17 of gestation, and numbers of corpora lutea, implantations and live and dead fetuses were recorded. After 4, 7 or 10 weeks of treatment, males were euthanized, genital organs were weighed, the number of sperm in the cauda epididymis was counted, sperm motion analyzed, and histopathological examination of testes performed. The male rats of the 1000 mg/kg dose group died or were moribund 3 or 4 days after the start of treatment. No effects on fertility of male rats were observed up to the 500 mg/kg 10-week dose group. Treatment for 4 weeks at 500 mg/kg/day decreased epididymis weight. After 7 weeks at 500 mg/kg/day, the weights of epididymis, seminal vesicles and prostate were decreased, and the number of sperm heads per cauda epididymis and percentage of motile sperm were reduced. In the 500 mg/kg 10-week dose group, the weight of testis was decreased. On histopathological examination of the testis, degeneration of seminiferous tubules and loss or exfoliation of spermatids were observed, and the ratio of retention of step 19 spermatids in stage IX-XI was increased in the 500 mg/kg 4-, 7- and 10-week dose groups. These results suggest that analysis of sperm motion and histopathological evaluation of testes are sensitive methods for assessing toxicity of valproic acid on male reproductive organs. PMID- 10845185 TI - Comedogenicity of squalene monohydroperoxide in the skin after topical application. AB - The comedogenicity of squalene peroxides was examined on the rabbit ear skin after topical application of squalene-monohydroperoxide (Sq-OOH), the initial product when squalene was irradiated with UV-A. Since comedogenic products from UV-irradiated squalene were extracted with methanol solution, we isolated Sq-OOH by reverse-phased HPLC with a methanol mobile phase solvent. The degree of comedogenic reaction induced by Sq-OOH was higher than that of well-known comedogenic cosmetic ingredients. Unlike two other mono-peroxides, tert-butyl hydroperoxide and cumene-mono-hydroperoxide, Sq-OOH induced comedo-formation in the rabbit ear skin. However, the comedogenicity of reduced Sq-OOH, squalene hydroxide (Sq-OH) and squalene itself was lower than that of Sq-OOH. These results indicate that Sq-OOH is a potent comedogenic mono-hydroperoxide chemical to rabbit skin. PMID- 10845187 TI - Spontaneous age-related peripheral neuropathy in B6C3F1 mice. AB - Peripheral neuropathy, which accompanies aging, occurs during the long-term rearing of laboratory animals. The present study set out to delineate the clinical and functional features of this neuropathy. A total of 200 B6C3F1 female mice, in groups of 5 to 20 mice, were sacrificed and autopsied each week beginning at 5 weeks and continuing to 130 weeks of age. Examination for histopathologic changes was conducted on the dorsal nerve roots, sciatic nerves, peroneal nerves, tibial nerves, plantar nerves and brachial nerve plexuses. At 90 weeks of age or later, peripheral neuropathy, characterized by axonal degeneration and Schwann cell proliferation, were observed mainly in the sciatic nerves, brachial nerve plexus and peroneal nerves. These spontaneous age-related nerve lesions appeared in all animals by 100 weeks of age in all nerves, and increased with increasing age. The nerve lesions were most prominent in the distal sciatic nerve. The rectal and hind-limb surface temperatures, motor nerve conduction velocity, blood glucose and HbA1C decreased with increasing age. Elevation of sorbitol contents in sciatic nerves and reduction of myo-inositol levels were also detected in 120-week-old mice. However, except for blood glycemic parameters, no correlation with peripheral nerve lesions could be demonstrated. Spontaneous hypoglycemia (< 40 mg/dL) persisted throughout the day in a small percentage (< 5%) of animals aged 80 weeks or more; these animals had extensive lesions in the peripheral nerves and showed decreased plasma levels of HbA1C and frucutosamines and increased plasma levels of ketones. These results suggest that spontaneous peripheral nerve disorders which accompany aging might worsen if spontaneous age-related hypoglycemia is also present. Such age-related changes must be taken into consideration in experimental studies performed on mice of this age. PMID- 10845188 TI - Toxicity and blood concentrations of xylazine and its metabolite, 2,6 dimethylaniline, in rats after single or continuous oral administrations. AB - To cast light on whether the carcinogenic risk of 2,6-dimethylaniline (DMA), a metabolite of xylazine, may increase by ingestion of edible tissues from domestic animals treated with xylazine, the following studies of xylazine and DMA were performed. In Experiment I, male F344 rats received a single oral administration of 150 mg/kg of xylazine hydrochloride. Rats showed symptoms suggesting loss of sensation and pain immediately after the treatment. These signs had disappeared after 3 hr, but the animals died of hydrothorax and pulmonary edema by 9 hr. The plasma concentration of xylazine was 2.88 +/- 0.95 micrograms/ml at 15 min, and then decreased to 0.10 +/- 0.01 microgram/ml at 6 hr. The plasma level of DMA remained at 0.03 to 0.04 microgram/ml during the measurement period. In Experiment II, male F344 rats were fed a diet containing 1000 ppm of xylazine hydrochloride, regarded as the maximum tolerated dose, for 4 weeks. No clear clinical signs were evident and the plasma levels of xylazine and DMA were at the detection limit (0.02 microgram/ml) or less, although follicular cell hypertrophy of the thyroid was observed in all the treated animals. In Experiment III, male F344 rats were fed a diet containing 3000 ppm or 300 ppm of DMA for 4 weeks. Histological changes, such as atrophy of Bowman's gland and irregular arrangement of olfactory epithelial cells, were only observed in the olfactory epithelium of the 3000 ppm group. The plasma levels of DMA were 0.20 to 0.36 microgram/ml in the 3000 ppm group, but under the detection limit in the 300 ppm group. These results suggest that the probability of nasal carcinogenic effects of DNA on consumers via ingestion of edible tissues from food-producing animals treated with xylazine is extremely low, since DMA levels in the blood of rats subjected to continuous administration of high doses of xylazine remained under the detection limit. PMID- 10845189 TI - Neonatal poisonings in middle Anatolia of Turkey: an analysis of 72 cases. AB - In this study, 72 newborn infants who were followed with the diagnosis of poisoning in Erciyes University Faculty of Medicine, Department of Pediatrics, Division of Neonatology, between 1975 and 1997 were evaluated retrospectively. Our purpose was to emphasize the importance of newborn poisoning among general poisoning in childhood. The age of infants ranged from 10 min to 25 days (0.82 +/ 2.81 days). Forty-seven (65.2%) infants were poisoned before or during delivery. Of the 47 infants' mothers, 46 had preeclampsia or eclampsia, and 27 received only magnesium sulfate; nine magnesium sulfate + diazepam; four magnesium sulfate + nifedipine; and the others received various drug combinations. Aside from these, one mother had Addison's disease and she used long-term dexamethasone during her pregnancy. In the newborn period, five (6.9%) infants inhaled organophosphate insecticides; eight (11.1%) ingested corrosive agents (four benzalkonium chloride; three chlorhexidine gluconate + cetrimide and an infant ammonium); four (5.5%) were poisoned by overdose of digoxin; three (4.1%) ingested overdose of phenobarbital; and two (2.7%) received acepromazine maleate. In addition, each infant ingested diphenoxilate HCL + atropine sulfate, pipenzolate bromid and tizanidine HCL. Follow-up period of the infants ranged from 24 hr to 26 days (0.82 +/- 2.81 days). The mortality rate was 17% (12/72). Death was not noted in the infants who were followed with poisoning after delivery. The causes of death were as follows: sepsis in four infants, meningitis, respiratory distress syndrome and necrotizing enterocolitis in two infants each, and the effects of overdose of magnesium sulfate and diazepam in two infants, respectively. In conclusion, we would like to stress that newborn infants whose mothers received magnesium sulfate or another drug during pregnancy or delivery should be closely monitored, and calculation of drug doses should be carefully taught to hospital nurses. When baby-rooms are disinfected with organophosphate insecticides in a hospital or house, infants should be removed from the room for at least 24 hr, and use of drugs should be explained in detail to the mothers. PMID- 10845190 TI - Thyroid hypertrophic effect of semotiadil fumarate, a new calcium antagonist, in rats. AB - We studied the effects of semotiadil fumarate (SF), a new calcium antagonist with a unique benzothiazine structure, on the thyroid gland and liver in rats and compared them with those of another calcium antagonist, nicardipine (NCD), a well known thyroidal hypertrophic agent and microsomal enzyme inducer, phenobarbital (PB), and goitrogen propylthiouracil (PTU). In oral 2-week treatment, SF caused increases in hepatic microsomal protein levels, uridine diphosphate glucuronosyltransferase (UDPGT) activity and an increase in serum thyroid stimulating hormone (TSH) level together with decreases in serum thyroid hormone levels. These results suggest that SF accelerates peripheral disposition of thyroid hormones and subsequently stimulates secretion of TSH from the pituitary gland as a compensatory response. PB and NCD had similar effects on the thyroid gland and the liver. PTU showed obvious thyroid hyperplasia and an increase in relative liver weight. Drastic increase in TSH level was observed in the PTU treated group together with significant decreases in serum thyroid hormone levels and an increase in hepatic UDPGT activity. Histopathologically, PTU depleted the colloids in follicles, suggesting the inhibition of thyroid hormone synthesis. SF, PB and NCD showed thyroidal hyperplasia, but the extent of the change was far more moderate than that induced by PTU. These results indicate that the effect of SF is similar to those of PB and thyroid hypertrophy seen in the oral 2-week treatment with SF, and may be caused by indirectly elevated TSH levels which resulted from the induction of hepatic UDPGT activity. PMID- 10845191 TI - Differentiation of responses to allergenic and irritant compounds in mouse popliteal lymph node assay. AB - The mouse popliteal lymph node assay (PLNA) has been proposed as an immunotoxicological test to predict allergenic chemicals. However, PLN response is also observed in association with non-specific activation induced by some irritants. We, therefore, examined the kinetics of the PLN cellularity indices of primary and secondary responses. Flow cytometric analysis was used to measure the proportions of T and B cells in PLN. Male ICR mice were subcutaneously injected with TNBS (an allergenic compound) or SDS (an irritant compound) in the left hind footpad, and with vehicle in the right one. On day 28 after first injection, mice were injected with 1/10 or 1/100 dose of the initial injection. On day 7 after first injection of TNBS, primary response reached maximal PLN cellularity index (16.5). On day 2 after second injection, secondary response reached maximal PLN cellularity index (13.1). A marked increase in proportion of B cells was observed in the PLN. On the other hand, after first injection of SDS, primary response reached maximal PLN cellularity index (2.8) on day 10, but neither secondary response nor increase in the B cell proportion were observed. These results suggest that the detection of secondary response and the flow cytometric analysis are useful in differentiating responses to allergenic and irritant compounds in the PLNA. PMID- 10845192 TI - The exacerbating effect of insulin-induced hypoglycemia on spontaneous peripheral neuropathy in aged B6C3F1 mice. AB - The effect of insulin-induced hypoglycemia on spontaneous peripheral neuropathy in aged mice was examined. Ninety-five-week-old female B6C3F1 mice were infused subcutaneously for 2 weeks with 40 or 80 IU/kg/day of insulin with a micro osmotic pump. Blood glucose level was decreased during the infusion (4.3-6.8 mmol/L in mice receiving 40 IU/kg/day of insulin or 2.4-5.4 mmol/L in mice receiving 80 IU/kg/day of insulin versus 6.5-7.6 mmol/L in control mice). In histopathological examination, axonal degeneration and/or remyelination were observed in a small number of nerve fibers of control mice. Similar nerve fiber lesions were observed in mice receiving 40 IU/kg/day of insulin, whereas severer lesions with an increase in segmental axonal degeneration of nerve fibers were observed in 4/7 mice receiving 80 IU/kg/day of insulin. These findings suggest that spontaneous peripheral neuropathy in aged mice is exacerbated by sustained hypoglycemia induced by insulin treatment. PMID- 10845193 TI - [X-ray and endoscopic diagnosis of esophageal achalasia]. AB - We evaluated diagnostic X-ray and endoscopic examination findings in 486 patients with esophageal achalasia. Concerning the association between the duration of disease and the X-ray dilatation type, the duration was 1-4 years in more than 50% of patients with the Sp type, a mean of 8.5 years in those with the F type, and frequently more than 10 years in those with the S type. Endoscopy is generally used to determine the presence or absence of abnormal movement. In achalasia, the endoscope can be inserted into the stomach despite resistance at the stenotic site, and the mucosal surface is normal. Squamous cell carcinoma as a complication was observed in 21 patients (4.3%). The carcinoma complication rate was higher with a longer duration of disease and a longer observation period. The mean total course including the postoperative course was 27 years. Long-term and periodic X-ray and endoscopic observation of the disease course is important, and iodine staining is indispensable for early detection of esophageal cancer. PMID- 10845194 TI - [Diagnosis using esophageal manometry and various loading tests]. AB - Two approaches of infusion and microtransducer manipulation are available for esophageal manometry. If esophageal peristalsis and relaxation of the lower esophageal sphincter (LES) are diminished, the diagnosis of achalasia can be made. As compared with the infusion method, the microtransducer method requires no perfusion of water and has no limit on posture, allowing successful measurement in an empty esophagus. Thus this method, which allows measurement after feeding and continuous monitoring for 24 hours, seems to be more physiological than the infusion method. With this method, however, peristasis like contractile waves and relaxation of the LES may be observed in addition to simultaneous contractile waves, even in cases of achalasia. Although methods to observe excessive reactions of the LES often involve a loading test with gastrin or mecolyl in some institutions, a loading test with cerulein is routinely used in our department. In healthy controls, administration of cerulein usually leads to decreased LES pressure, while increased LES pressure is observed in patients with achalasia (paradoxical response). PMID- 10845195 TI - [Motility disorders of the esophagus, excluding achalasia]. AB - Esophageal motor disorders can be primarily focused in the esophagus or secondarily related to a systemic disease. Based on the manometric findings, primary esophageal motor disorders can also be classified into achalasia and spastic disorders, such as diffuse esophageal spasm, nutcracker esophagus, hypertensive lower esophageal sphincter, and nonspecific esophageal motility disorders. PMID- 10845196 TI - [Clinical management of esophageal achalasia]. AB - In this report, we describe the clinical management of esophageal achalasia in Japan. Both balloon dilation and surgical myotomy or myectomy with antireflux procedures are accepted as the standard treatment for esophageal achalasia. In most patients balloon dilation is attempted first. If that is not sufficiently effective, they than undergo surgery. The most important point in treatment is not to repeat the dilation procedure more than twice, because patients who do not respond significantly to the first two dilation procedures are unlikely to improve after subsequent ones. Recently, many physicians and surgeons in Western countries have investigated the effects of botulinum toxin in patients with esophageal achalasia. If the safety of this treatment is confirmed in the near future, it will become a new treatment option for esophageal achalasia. PMID- 10845197 TI - [Esophageal dilation therapy for achalasia of the esophagus]. AB - Esophageal dilation therapy for esophageal achalasia has a 300-year history and remains the first-line therapy because of its low cost, progressively decreasing morbidity and mortality, and possibility of surgery for patients who do not improve. Another advantage is the feasibility of repeated trial even after surgery. The aim of this procedure is to produce a controlled tear in the lower esophageal sphincter muscle. Precise esophagographic, endoscopic, and manometric diagnosis is necessary for successful esophageal dilation therapy. In addition, the exclusion of pseudoachalasia using echogram or CT scan is important. The procedure should be carried out accurately to prevent complications. If perforation occurs, appropriate, timely management is required. PMID- 10845198 TI - [Laparoscopic surgery for esophageal achalasia]. AB - Laparoscopic surgery for esophageal achalasia was first reported by Shimi et al. in 1991. Subsequently the procedure has been performed all over the world and laparoscopic Heller myotomy and Dor fundoplication (Heller and Dor operation) is now thought to be the operation of first choice. It is indicated for patients who are resistant to medical therapy (calcium blocker etc.) or have pneumatic dilatation and those with frequent aspiration at night. As Csendes et al. reported that surgical treatment was better than pneumatic dilatation and as laparoscopic surgery is less invasive, the indications for the laparoscopic Heller and Dor operation can include all achalasia patients except those who respond to medical therapy, do not accept surgery, or cannot tolerate surgery. We successfully performed the laparoscopic Heller and Dor operation on 22 patients, all of whom had an uneventful postoperative course. Manometric evaluation, endoscopic examination, and 24-hour pH monitoring showed good results. There are six important technical points: 1) flexible laparoscopy; 2) pneumoperitoneum; 3) gauze in the abdominal cavity to absorb blood; 4) laparosonic coagulating shears; 5) extracorporeal knot-tying technique; and 6) intracorporeal knot-tying technique. If an experienced surgeon is in charge, the laparoscopic Heller and Dor operation is an ideal, minimally invasive treatment for esophageal achalasia. PMID- 10845199 TI - [Modified Jekler and Lhotka operation in minimized laparotomy for achalasia]. AB - The rationale for surgery in achalasia is to reduce lower esophageal sphincter (LES) pressure and recontruct an antireflux mechanism. Fundopexy as an antireflux procedure after LES myotomy (modified Heller myotomy), called the Jekler and Lhotka method or the Heller and Dor operation, is commonly performed as standard surgery. In our institution, this method has been used since 1982. We have performed the modified Jekler and Lhotha method including long myotomy to confirm complete reduction of LES pressure as measured by intraoperative manometry and fundopexy plus posterior wall fixation to contract the long abdominal esophagus since 1986. Satisfactory results with this procedure have been obtained and reported. Furthermore, we have recently performed this procedure with minilaparotomy. In this review, we describe the modified Jekler and Lhotha method in minimized laparotomy and evaluate this procedure using intraoperative manometry. PMID- 10845200 TI - [Recent advances in the surgical management of esophageal achalasia: modified Girard method and laparoscopic approach]. AB - The standard surgical treatment for esophageal achalasia consists of contraction of the lower esophageal sphincter combined with antireflux repair. This paper describes the techniques of the modified Girard method with a laparoscopic approach. The modified Girard method consists of a long esophagomyotomy, the placement of horizontal transverse sutures to prevent restricture due to reflex, and fundopexy for antireflex repair. We have recently used a laparoscopic approach with the modified Girard method in 10 patients with esophageal achalasia. A total of five trocars were required for the long myotomy in the lower esophagus and repair of the esophagogastric junction. A myotomy of approximately 10-12 cm was made along the lower esophagus and cardia lesion using an electric J-hook. Than transverse sutures were placed on each side of the lower myotomy and the gastric fundus was covered with the upper myotomy with several sutures to diaphlagm and each side of the myotomy lene. Excellent results were achieved, and the patients recovered from surgery more quickly compared with those undergoing an open procedure. However, postoperative manometric findings did not differ between patients who underwent the laparoscopic and open procedures. PMID- 10845201 TI - [Improved gastric patch method for esophagocardioplasty in esophageal achalasia]. AB - We performed esophagocardioplasty with the gastric patch (original GP) as the treatment of first choice in patients with esophageal achalasia. However, postoperative reflux esophagitis occurred in many due to secretory dysfunction of the gastric mucosa implanted in the mediastinum. We therefore developed two modifications of the gastric patch technique (GP II and GP III) which do not involve implantation of the gastric mucosa in the esophagus. Twenty-three patients have undergone the modified GP II procedure in our institution. Postoperative changes in the esophagus were examined fluoroscopically, and it was found that esophageal diameter increased from an average 2.9 +/- 0.6 cm to 5.2 +/ 1.0 cm at an average follow-up of 7.1 months (p < 0.01). All 23 patients experienced amelioration of symptoms, even though some reddening was still present in the lower esophagus, with a maximum follow-up of 8 years. Based on these results, the modified GP procedures are an acceptable open surgery modality for esophagocardioplasty in esophageal achalasia. PMID- 10845202 TI - [Aortocaval fistula with acute right heart failure during surgery: case report]. AB - A 71-year-old man was transferred to our institution with suspected of impending rupture of the abdominal aorta. The clinical manifestations included abdominal pain, pulsatile mass, and abdominal bruit. We made on early diagnosis of aortocaval fistula based on enhanced computed tomography and color Doppler flow imaging. Surgical repair was performed 24 hours after the onset of symptoms. At the introduction of anesthesia, cardiac arrest with high central venous pressure occurred. After resuscitation and rapid clamping of the proximal aorta, surgical closure of the fistula with graft replacement was performed successfully. PMID- 10845203 TI - [Clinical significance of stroke care unit]. PMID- 10845204 TI - [Sequential nerve conduction studies in a patient with ulnar neuropathy at the elbow treated by night athletic supporter]. AB - Ulnar nerve can be stretched with the elbow flexed position. To avoid elbow flexed position in patients with ulnar neuropathy at the elbow we used an athletic elbow supporter. We herein demonstrate a 31-year-old man with right ulnar neuropathy at the elbow whose neuropathy was resolved by using this supporter only at night. He had complained of weakness and paraesthesia in the ulnar side of his right hand. Nerve conduction studies of right ulnar nerve revealed decrease in the amplitude of compound nerve action potentials and a severe motor nerve conduction block with apparent conduction delay around the ulnar groove. A diagnosis of ulnar neuropathy at the elbow was done and we recommended him to wear an athletic elbow supporter at night. Paraesthesia of his right hand improved in a few days after starting this therapy. Three months later paraesthesia was resolved. One year later grip power of his right hand increased to 35 kg from 20 kg, and the conduction block at the elbow completely disappeared. Compound nerve action potentials, recorded at the segment of wrist to above elbow and wrist to finger, were improved equally. These observations suggest that the conduction block at the elbow entrapment site and the distal axonal degeneration gradually recovered together. PMID- 10845205 TI - [Surgical treatment for temporal lobe epilepsy in childhood]. AB - We retrospectively analyzed 5 children (11-15 year) with intractable temporal lobe epilepsy (TLE) who underwent the anterior temporal lobectomy with hippocampectomy. Cases 1-3 had medial TLE (MTLE) with histologically verified hippocampal sclerosis, Case 4 had lateral TLE, and Case 5 had MTLE with old hemorrhagic lesion in the lateral temporal lobe. In Cases 3-5, chronic invasive electrocorticography recording using subdural electrodes was obtained, while in Cases 1 and 2, the epileptogenic region was defined by noninvasive preoperative evaluation. Postoperatively, Cases 1-3 became seizure free. All patients had psychosocial problems after the onset of their epilepsy, which was not improved even after the surgical control of epilepsy. Since most patients had morphological change and perfusional and metabolic disturbance outside the hippocampus at the time of surgery, earlier surgical consideration may be necessary. PMID- 10845206 TI - [Medulloblastoma with neuronal differentiation: a report of five cases]. AB - This report presents a retrospective analysis of 5 patients who were treated for cerebellar medulloblastoma with neuronal differentiation. Four males and 1 female ranged in age from 6 months to 9 years at the time of diagnosis. Total removal of the tumor was achieved in 3 patients, and partial removal in 2. While these tumors were composed of small cells and had regions resembling desmoplastic medulloblastoma, they in part showed neuronal characteristics which included parallel row or linear array arrangements of tumor cells in an eosinophilic fibrillary matrix. Postoperatively, 3 patients received craniospinal radiation therapy, one received local radiation to the primary site, and the remaining one received only systemic chemotherapy. During the follow-up period of 3.8-25.2 years, 4 patients have been in continuous remission with mild to moderate neurological deficits, while the remaining one died 3.9 years after surgery. The clinical and anatomic pathological features of medulloblastomas with neuronal differentiation are reviewed while the therapeutic problems associated with these tumors are also discussed. PMID- 10845207 TI - [A case of chronic toluene intoxication presenting stimulus-sensitive segmental spinal myoclonus]. AB - We reported a 48-year-old male who showed stimulus-sensitive spinal myoclonus due to chronic toluene intoxication. He has been exposed to thinner for more than 30 years as a painter, and occasionally experienced an episode of headache, nausea and dizziness because of acute thinner intoxication. He noted tremor of his hands 10 years ago. He also noticed memory disturbance since the end of 1997. Neurological examination revealed postural tremor of his fingers on the bilateral sides and the left arm. In addition, rhythmic myoclonic jerks were induced in the right upper limb muscles by a tendon tap given on the right brachioradialis muscles. Surface EMG revealed repetitive grouping discharges in those two muscles approximately 100 msec after the tendon tap which continued for about 30-50 msec. A long loop reflex (C-reflex) and giant SEPs were not observed in his right upper limb, and EEG showed no spike. Urinary excretion of N-benzoylglycine, which was a metabolite of toluene was increased (1.17 g/l). Therefore, he was diagnosed as a case of chronic toluene intoxication. His myoclonic jerks were considered to be stimulus-sensitive spinal myoclonus, because they were induced segmentally and because cortical hyperexcitability was not seen. This is the first report to describe the occurrence of stimulus-sensitive spinal myoclonus in the case of chronic toluene intoxication. PMID- 10845208 TI - [Initial positive deflection of the compound muscle action potential in the median nerve conduction studies can be originated from lumbrical muscles in patients with carpal tunnel syndrome]. AB - In motor nerve conduction studies we sometimes encounter a small initial positive deflection (IPD) of the compound muscle action potential (CMAP). This potential represents a volume conduction from nearby muscles other than the objective muscle. We demonstrated recordings of motor nerve conduction studies from two patients with carpal tunnel syndrome (CTS). In patients with CTS IPDs can be recorded from a surface electrode above the abductor pollicis brevis when intense stimuli to the median nerve provoked a stimulus spread to the ulnar nerve. However, without this stimulus spread to the ulnar nerve, IPDs can be observed by contraction of median nerve innervated muscles. In the CTS thenar branch of the median nerve is apt to be more severely damaged than lumbrical branch. In such an occasion volume conduction from the lumbrical muscles is relatively large, which gives rise to the IPD in the CMAP recorded from abductor pollicis brevis. We reported two cases of IDPs originated from lumbrical muscles. The peak latencies were identical between IDP of abductor pollicis brevis recording and negative potential of lumbrical recording. These potentials didn't change by median nerve stimulation at the elbow 3 msec after the ulnar nerve stimulation at the wrist (collision technique). Finally, we repeat that IPDs in the median nerve conduction studies can be originated from not only the stimulus spread to the ulnar nerve but also the median nerve innervated lumbrical muscles in patients with CTS. PMID- 10845209 TI - [A case of multiple cranial neuropathy with positive antinuclear antibody responded to steroid]. AB - A 56-year-old woman was admitted to our hospital because of bilateral ptosis, total ophthalmoplegia, bilateral facial palsy and left hypoglossal nerve palsy. Antinuclear antibody (ANA) showed high titer of 1280 but other data were normal. With oral prednisolone therapy (40 mg/day), the symptoms improved gradually but ANA titer did not show any significant change. We thought multiple cranial neuropathy of the patient was due to autoimmune disease such as collagen disease or cranial nerve type of Guillain-Barre syndrome from her symptoms and positive ANA. In contrast to other reported cases of multiple cranial neuropathy with positive ANA, in our patient ANA titer was high, only motor cranial nerves were impaired, and total ophthalmoplegia was found. Steroid therapy was useful in accordance with other reports. Although our patient has no symptoms related to collagen diseases, a long follow-up of this patient is considered to be necessary. PMID- 10845210 TI - [Radiation induced glioblastoma: a case report]. AB - We report a surgical case of a 54-year-old woman with a radiation induced glioblastoma. At the age of 34, the patient was diagnosed to have a non functioning pituitary adenoma. It was partially removed followed by 50 Gy focal irradiation with a 5 x 5 cm lateral opposed field. Twenty years later, she suffered from rapidly increasing symptoms such as aphasia and right hemiparesis. MRI showed a large mass lesion in the left temporal lobe as well as small mass lesions in the brain stem and the right medial temporal lobe. These lesions situated within the irradiated field. Magnetic resonance spectroscopy revealed relatively high lactate signal and decreased N-acetyl aspartate, choline, creatine and phosphocreatine signals. Increased lactate signal meant anaerobic metabolism that suggested the existence of a rapidly growing malignant tumor. Thus, we planned surgical removal of the left temporal lesion with the diagnosis of a radiation induced malignant glioma. The histological examination revealed a glioblastoma with radiation necrosis. MIB-1 staining index was 65%. Postoperatively, her symptoms improved, but she died from pneumonia 1 month after the surgery. An autopsy was obtained. The lesion of the left temporal lobe was found to have continuity to the lesion in the midbrain, the pons and the right temporal lobe as well. High MIB-1 staining index suggested that a radiation induced glioblastoma had high proliferative potential comparing with a de novo and a secondary glioblastoma. PMID- 10845211 TI - [Usefulness of ultrasonography in a case of spontaneous carotid-cavernous fistula: a case report]. AB - Since spontaneous carotid-cavernous fistula (CCF) is a dural arterio-venous fistula at the cavernous sinus, which is different from traumatic CCF and CCF associated with a ruptured aneurysm at the cavernous internal carotid artery, cerebral angiography is required in order to differentiate these condition. We here report a case of spontaneous CCF, in which a result of ultrasonographic evaluation of cervical arteries well corresponded with that of cerebral angiography. Ultrasonography showed increased blood flow and decreased pulsatility index in the ipsilateral external carotid artery, contralateral internal and external arteries, and these values in all arteries resolved within normal range after the interventional embolization. Ultrasonography is less invasive examination and can be easily performed even in outpatients for observation of spontaneous CCF. PMID- 10845212 TI - [L-dopa effective parkinsonism appeared after shunt revision of the aqueductal stenosis: report of two cases]. AB - We present a two cases of parkinsonism that appeared after shunt revision in obstructive hydrocephalus patients. Case 1 was a 59-year-old man and case 2 was a 32-year-old woman. Parkinsonism appeared in both cases after 3 or 4 shunt revisions within a period of three months. Treatment with bromocriptine, amantadine and L-dops had no effect, but the parkinsonism dramatically improved with administration of L-dopa. It was thought that repeated dilatation and collapse of the third ventricle in a short period of time might have caused concussion of the nigrostriatal pathway, resulting in reversible function loss of the nigrostriatal pathway. PMID- 10845213 TI - [A case of juvenile bow hunter's stroke]. AB - Bow hunter's stroke results from vertebrobasilar insufficiency caused by mechanical occlusion or stenosis of the vertebral artery at the C 1-2 level on head rotation. Commonly it is seen in elder people with cervical spondylosis. Here we reports a case of bow hunter's stroke in a 25-year-old male who complained of visual disturbance and syncope on rotation of the head 90 degrees or more to the left. This problem was frequently seen on driving a car. A cervical x-ray and MRI of the head revealed no abnormal findings such as atlantoaxial dislocation. Angiograms demonstrated obstruction of the right vertebral artery at the C 1-2 level on left rotation of the head. The hemodynamics on the circle of Willis were evaluated and surgical treatment was planned. The posterior fusion involving C 1-2 has long been used to limit atlantoaxial rotational movements. However, it has the serious disadvantage because the range of head motion is severely reduced. Recently decompression of the atlantoaxial portion of the affected vertebral artery has been used, but recurrence of occlusion with head rotation can be seen postoperatively. Since the patient could predict the onset of attack, we managed him conservatively, and no traffic accident reported during this period of observation. Due to several surgical disadvantages, we propose that whenever possible, patients with bow hunter's stroke should be managed conservatively especially in young patients. PMID- 10845214 TI - [A long-term follow-up case of intrahypothalamic type of hypothalamic hamartoma]. PMID- 10845215 TI - [Myxopapillary ependymoma]. PMID- 10845216 TI - [Hyperostosis of the cervical spine presenting as myelopathy: a case report]. PMID- 10845217 TI - Justification and controllable dose. PMID- 10845218 TI - 57Co-EDTA renal imaging in rats. AB - We studied the synthesis of 57Co-EDTA (Ey = 122 keV), its biodistribution in Wistar rats and its blood and urinary elimination compared with that of 51Cr EDTA. We added 6 mumol EDTA diluted in 3-5 ml isotonic phosphate buffer (Na2HPO4) to a commercial 57CoCl2 radioactive tracer solution. The incubation period was 15 min. Quality control was performed using TLC and HPLC. Six healthy Wistar rats underwent 57Co-EDTA renography for 30 min. In one rat, additional TLC and HPLC was performed on blood (one sample only) and urine samples (n = 3) obtained 30 min, 30 min, 2 h and 4 h following injection of 18.5 MBq 57Co-EDTA and 51Cr-EDTA respectively. Radioisotope quantification was done by means of a germanium detector. 57Co was chelated to EDTA at high yield (Kstab = 10E36). No free or protein-bound 57Co was found. The ratio of 51Cr-EDTA to 57Co-EDTA remained constant (P = 0.133, n = 4). 57Co-EDTA was rapidly cleared from the blood pool (heart), and prompt and high target-to-background ratios for both kidneys were obtained (mean = 8.4, range = 7-12). At the end of the acquisition, activity remaining in the body excluding kidney and bladder was 45 +/- 5.2%. No specific activity uptake was noted in any other organ or tissue. We conclude that 57Co EDTA is a promising radioligand for simultaneous clearance and separate renal function estimation. Its preparation is straightforward and, in rats, no free or protein-bound 57Co was found. PMID- 10845219 TI - NORA: a simple and reliable parameter for estimating renal output with or without frusemide challenge. AB - The aim of this study was to evaluate a simple parameter describing renal output, namely NORA (normalized residual activity). We first compared, in a simulated model, different parameters of transit to an ideal standard; we then compared, in a clinical study, NORA and output efficiency. 123I-hippurate, 99Tcm-DTPA and 99Tcm-MAG3 plasma curves, each with two levels of renal clearance, were convoluted by means of different types of simulated retention functions, with different mean transit times. On the reconstructed renograms, several parameters reflecting renal transit were determined and compared with mean transit time. In a second step, in 33 patients, we compared output efficiency and NORA (i.e. the residual renal activity), normalized by the renal activity at 2 min. These two parameters were calculated at the end of the renogram, at the end of the frusemide test and after the micturition phase. In the simulated model, both output efficiency and NORA were only slightly influenced by the level of overall renal function. In the clinical study, a good correlation was found between output efficiency and NORA, whatever part of the study considered (renogram, frusemide test, post-voiding image). NORA is a simple and reliable parameter that allows quantification of renal output; it is almost independent of the level of renal function and can be used whatever the timing of the frusemide injection. PMID- 10845220 TI - Aspirin renography in the diagnosis of renovascular hypertension: a comparative study with captopril renography. AB - Captopril renography is used for the non-invasive diagnosis of renovascular hypertension, but suffers from the drawbacks of lower sensitivity and false positive tests due to a fall in blood pressure. Aspirin renography has been proposed as a useful test for evaluation of unilateral renal artery stenoses of moderate degree. We studied the clinical usefulness of aspirin renography in 12 patients with a clinical suspicion of renovascular hypertension and compared it with captopril renography using 99Tcm-DTPA. The test was considered positive if there were changes in the time-activity curve according to the criteria specified by the American Society of Hypertension Working Group. Four patients with discordant results between captopril and aspirin underwent intra-arterial digital subtraction angiography. In two patients, the renal arteries were normal; captopril was false-positive in both these patients. Bilateral stenosis was noted in the third patient, with captopril being false-negative on the right side with moderate stenosis, whereas aspirin was true-positive. There was unilateral stenosis in the fourth patient; captopril was false-positive on the contralateral side. Our results suggest that aspirin renography is superior to captopril renography in the assessment of patients with a suspicion of both unilateral and bilateral renovascular hypertension. PMID- 10845221 TI - The role of 99Tcm-tetrofosmin in the evaluation of thyroid nodules. AB - Various radionuclides, including 67Ga, 201Tl and 99Tcm-sestamibi, have been used to differentiate benign from malignant thyroid nodules. 99Tcm-tetrofosmin, a lipophilic cationic radiotracer, and 99Tcm-sestamibi have also been reported to accumulate in thyroid tumours. In this study, we evaluated the role of 99Tcm tetrofosmin in the differentiation of malignant from benign thyroid nodules. Seventy-nine patients with solitary non-functioning thyroid nodules were included in the study. Fine-needle aspiration biopsy was performed in all patients. Sixty patients were subsequently operated on and 19 patients refused surgery. After the injection of 370 MBq 99Tcm-tetrofosmin, static images at 5, 30, 60, 120 and 180 min were acquired. Both visual and semi-quantitative analysis was performed. On visual interpretation, the nodules with late retention were classified as positive for malignancy and nodules without late retention were classified as negative for malignancy. In the semi-quantitative analysis, regions of interests were drawn over the nodule and contralateral normal thyroid tissue. The average number of counts was recorded and tumour-to-normal thyroid tissue ratios calculated. Post-operative histology revealed 19 malignant and 41 benign nodules. Of the benign nodules, adenomas behaved similarly to the malignant nodules with late retention of tracer, while adenomatous nodules revealed no late retention on delayed images and could be differentiated from malignant tumours. In the semi quantitative analysis, there was a significant difference in tumour-to-normal tissue ratios for adenomatous nodules and malignant tumours as well as adenomas. We conclude that it is not possible to differentiate between malignant and benign thyroid nodules with 99Tcm-tetrofosmin. However, 99Tcm-tetrofosmin scintigraphy is helpful in selecting nodules that can be cured by surgical intervention. PMID- 10845222 TI - The value of early and double phase 99Tcm-sestamibi scintimammography in the diagnosis of breast cancer. AB - The aim of this study was to assess the additional value of early and double phase scintimammography (SMM) with 99Tcm-sestamibi in the detection of breast cancer following initial evaluation by palpation and mammography. Altogether, 322 women with breast lesions evaluated prospectively by palpation, fine-needle aspiration and mammography were assigned a malignancy risk according to the results. Scintimammography was performed in all patients in the prone breast dependent position. Immediate and delayed views were obtained. Acquisition of immediate tracer uptake was termed 'early phase' SMM, whereas a combination of both immediate and delayed phase images was termed 'double phase' SMM. All patients underwent breast biopsy. Both early phase and double phase SMM detected eight of nine tumours in the low-risk group (88.8% sensitivity). In the uncertain cases (moderate-risk group), early phase SMM detected all malignant tumours, but double phase SMM missed one (92.3% sensitivity). In the high-risk group, early phase SMM missed two breast cancers (94.6% sensitivity) and double phase SMM missed four (89.2% sensitivity). Overall, early phase SMM had a sensitivity of 94.9% and a specificity of 80.2% in detecting breast cancer, whereas double phase SMM had a sensitivity of 89.8% and a specificity of 94.3%. Both methods had 100% sensitivity for tumours larger than 1 cm. In conclusion, SMM detected additional breast cancers following an initial evaluation by palpation, fine-needle aspiration and mammography. Our results suggest that double phase SMM is more specific than early phase SMM, although early phase SMM is more sensitive. Whether the interpretation of SMM results should rely on both early and delayed images, or early images alone, should be based on the relative risk of malignancy of the breast lesion as estimated by the initial evaluation. PMID- 10845223 TI - Radiochemical purity of routinely prepared 99Tcm radiopharmaceuticals: a retrospective study. AB - Radiochemical purity is an important quality parameter for radiopharmaceuticals. In this study, the radiochemical purity of 2090 samples out of 7000 routine preparations of 20 different 99Tcm radiopharmaceuticals was tested using standard methods over a period of more than 7 years. The mean radiochemical purity was 96.92% (standard deviation = 6.71%). Seventy-four preparations failed to meet radiochemical purity limits; that is, 3.54% of all preparations tested or 1.06% of all preparations in the observation period. The reasons for substandard preparations were mainly related to laboratory-specific conditions. The introduction of a dedicated quality control protocol allowed the elimination of many sources of labelling failures and could reduce the number of administered preparations with an insufficient radiochemical purity. We stress the need for quality control in the preparation of radiopharmaceuticals and provide original radiochemical purity values of routinely prepared 99Tcm radiopharmaceuticals. PMID- 10845224 TI - Absorbed dose estimates for 131I-labelled monoclonal antibody therapy in patients with intraperitoneal pseudomyxoma. AB - Seven patients with intraperitoneal pseudomyxoma originating from the appendix (4 cases) and from the ovary (3 cases) were treated with radioimmunotherapy. During the therapy, nine infusions of 3.0-4.2 GBq of 131I-labelled B72.3 monoclonal antibody were administered. We developed three-dimensional dose calculation software that can utilize activity maps based on SPET images to calculate the absorbed dose distribution using point source kernels. The dose calculation program was employed to calculate absorbed doses to various organs. The calculated dose distributions enable us to evaluate the variation in dose within the organs, which is normally not available using approaches based on geometric models. The patient-specific absorbed dose calculations were compared with doses based on a model that uses photon S-factors derived from a standard phantom. The compared doses agreed well on average, but in some organs showed large discrepancies. PMID- 10845225 TI - Time for abstention from caffeine before an adenosine myocardial perfusion scan. AB - Seventy patients undergoing adenosine myocardial perfusion scintigraphy were studied. All patients reported abstention from products containing caffeine in the 12 h prior to the test. Blood samples were drawn prior to initiation of the stress test, and serum caffeine levels were determined using high-performance liquid chromatography. All patients were also asked about their coffee and tea drinking habits. Seventy-four percent of patients had measurable serum caffeine levels (n = 52) ranging from 0.1 to 8.8 mg.l-1. Results were correlated with maximum pulse rate, systolic and diastolic blood pressure changes and clinical symptoms during the test. There was no correlation between coffee or tea drinking habits and serum caffeine levels. A serum caffeine level of 2.9 mg.l-1 was considered a cut-off point for comparing patients. No significant difference was seen in mean maximum change of pulse rate, systolic and diastolic blood pressure between patients with serum caffeine levels > or = 2.9 mg.l-1 and those with lower serum caffeine levels. Of eight patients with serum caffeine levels > or = 2.9 mg.l-1, six had no symptoms (75%). When patients were classified as patients with no symptoms or patients with symptoms (mild, moderate or severe), a significant difference was demonstrated between patients with serum caffeine levels > or = 2.9 mg.l-1 and those with lower levels. This suggests 12 h abstention from caffeine may be insufficient. Whether this translates into false negative perfusion scans should be the subject of a larger study. PMID- 10845226 TI - Umbilical cord blood transplantation in beta-thalassemia major. PMID- 10845227 TI - Prepubertal growth and growth hormone secretion in children after treatment for hematological malignancies, including autologous bone marrow transplantation. AB - Prepubertal growth standards were used to assess growth in 20 children who had undergone autologous bone marrow transplantation (ABMT) as part of their treatment for hematological malignancy. Most of the patients (16 of 20) were transplanted after a relapse of their disease. A negative change in height standard deviation score (H-SDS) was seen only in the group of patients (n = 7) who had received both cranial irradiation therapy (CRT) and 7.5-Gy single fraction total body irradiation (TBI). Height changes in this group were observed from the time of diagnosis. In contrast, the groups of patients conditioned with chemotherapy only (n = 3) or both chemotherapy and TBI, without preceding CRT (n = 10), did not demonstrate a significant loss in H-SDS. Weight related to height demonstrated large individual differences over time. Spontaneous growth hormone (GH) secretion, as measured by a four-point sleep curve, was followed longitudinally and an increasing proportion of patients with low peak levels was seen in all patient groups. In summary, prepubertal growth was suppressed only in patients who received cranial irradiation before ABMT. Despite low GH peak levels, normal prepubertal growth was found in patients with no CRT before ABMT. PMID- 10845229 TI - Combined umbilical cord blood and bone marrow transplantation in the treatment of beta-thalassemia major. AB - The authors report on three children with beta-thalassemia major, class II, III, and III according to the Pesaro classification, with a body weight of 16, 62, and 50 kg, respectively, who received grafts using both umbilical cord blood (UCB) and bone marrow (BM) stem cells from their HLA-matched siblings. The number of UCB nucleated cells collected was 2 x 10(7)/kg, 1.2 x 10(7)/kg, and 2.5 x 10(7)/kg, respectively, and was considered insufficient to secure engraftment. The authors increased the number of hematopoetic progenitors by harvesting BM from the same donors. All 3 patients showed prompt engraftment with neutrophil recovery on days 17, 18, and 17 post-transplant, respectively, and platelet recovery on days 19, 25, and 22 post-transplant, respectively. One patient had remarkably increased HbF of values 31, 19, and 12% at 3, 6, and 12 months post transplant, respectively, which were accompanied by an increase in the G gamma/A gamma ratio, suggesting UCB-derived hematopoetic reconstitution. All patients are alive and transfusion independent 23, 18, and 16 months post-transplant, respectively. For patients with homozygous beta-thalassemia who are at high risk of graft failure, either because of major prior alloimmunization or an insufficient amount of UCB stem cells, combined transplantation with UCB and BM could offer a quick and safe alternative therapy. PMID- 10845228 TI - Thrombocytopenia with absent radii: frequency of marrow megakaryocyte progenitors, proliferative characteristics, and megakaryocyte growth and development factor responsiveness. AB - Congenital thrombocytopenia with absent radii (TAR syndrome) is characterized by defective thrombopoiesis and bleeding in early infancy. To determine the frequency and responsiveness to cytokines of megakaryocyte progenitors (CFU-Meg) in TAR syndrome, the authors studied marrow samples from 3 patients and 6 normal controls, using optimally standardized megakaryocyte growth media incorporating interleukin-3, interleukin-6, stem cell factor, and granulocyte-monocyte colony stimulating factor, with and without pegylated recombinant human megakaryocyte growth and development factor (PEG-rHuMGDF). CFU-Meg was identified with a specific staining system utilizing monoclonal antibodies to glycoprotein IIb/IIIa. Growth of small CFU-Meg colonies (3-20 cells/colony) was observed in all patients in cultures without PEG-rHuMGDF, with a mean frequency of 8 (range 5 12) per 2.25 x 10(5) mononuclear cells plated (control mean 23; range 2-70). Identical cultures of marrow cells from patients and controls with added PEG rHuMGDF produced more colonies per dish (mean 17, range 8-23; control mean 30, range 6-62). Except for 1 case, however, patients' colonies in response to PEG rHuMGDF remained smaller than those of controls. Two patients tested had higher plasma thrombopoietin levels than 6 normal subjects. The findings demonstrate proliferative and PEG-rHuMGDF-responsive megakaryocytic progenitors in TAR syndrome. The modest reduction in frequency of megakaryocyte progenitors and the suboptimal size of colonies in response to PEG-rHuMGDF are compatible with the reported defective signal transduction in the c-mpl pathway in TAR syndrome. PMID- 10845230 TI - Spontaneous apoptosis and retinoic acid receptor incidence in neuroblastomas and peripheral neuroectodermal tumors. AB - Twelve cases of neuroblastoma (NB) (7 boys and 5 girls) and 4 cases of primitive peripheral neuroectodermal tumor (PNET) (3 boys and 1 girl) were investigated for the presence of apoptosis and retinoic acid receptor (RAR) by immunhistochemical method. The apoptotic index in NB was zero or 1% in 8 children and relatively low (2-4.8%) in the other 4 cases, while it was higher (4.1-10.5%) in PNET. The RAR index determined by immunoperoxidase reaction in NB was zero or 3% in 5 cases and 9-34% in 7 children. RAR index in PNET was 16-68% in all the 4 cases. Good correlation (r = .47 according to Pearson-Bravis) was found between the number of RAR and spontaneous apoptosis. These results suggest that the RAR index in untreated NB and PNET shows great individual variation since its determination is necessary for the evaluation of the efficacy of retinoic acid treatment. PMID- 10845231 TI - Hemophagocytosis complicating Kawasaki disease. AB - A 6-year-old boy developed hemophagocytic syndrome during the recurrent course of Kawasaki disease. Despite the appropriate treatment modalities for Kawasaki disease, he developed pancytopenia, marked hepatosplenomegaly, high-grade fever, hyperferritinemia, hypertriglyceremia, and evidence of hemophagocytosis in the liver biopsy. Although the course was stormy, he responded well to a combination therapy of corticosteroids, etoposide VP16, and granulocyte colony-stimulating factor G-CSF. The clinical course and the treatment given were compared with the previous reported cases. PMID- 10845232 TI - Recurrent urinary tract infections and genitourinary tract abnormalities in the Imerslund-Grasbeck syndrome. AB - Two Imerslund-Grasbeck patients who presented with recurrent urinary tract infections and genitourinary abnormalities are described. The patients were evaluated with abdominal ultrasounds, voiding cystourethrograms, and Schilling tests. Each patient had large postvoid residual urine secondary to a motor neurogenic bladder. One had a duplication of the distal urethra manifesting as two meatal openings. There was lack of urinary excretion of radioactive vitamin B12 on Schilling tests in both patients. Patients with Imerslund-Grasbeck syndrome may be predisposed to urinary tract infections because of incomplete bladder emptying. Complete physical and radiological examinations of the genitourinary tract should be performed. PMID- 10845233 TI - Immediate free flap mandibular reconstruction in osteosarcoma of the mandible in childhood. AB - Mandibular osteogenic sarcoma (OS) is a very rare entity in childhood. Adequate surgical rejection with a wide margin of normal tissue is the mainstay of treatment of this site, while the role of adjuvant chemotherapy remains uncertain. A case is presented of a 15 1/2-year-old male with a huge OS of the mandible. The boy underwent surgical resection of the mandible with immediate fibula free flap reconstruction and is alive and free of disease 6 1/2 years following unitial diagnosis. This case suggests that immediate bone reconstitution with vascularized grafts have good functional and morphological results for osteosarcoma of the lower jaw. PMID- 10845234 TI - Primary T-cell lymphoma of the brain in children: a case report and literature review. AB - Described here are the clinical features and results of treatment in a 10-year old Saudi Arabian girl with primary T-cell lymphoma of the central nervous system. At presentation the patient had nystagmus and ataxia. The diagnosis was established by tissue biopsy obtained from the cerebellum. Therapy included cranio-spinal irradiation and combination chemotherapy of a systemic high dose of methotrexate, cytosine, arabinoside, and L-asparaginase. Remission was obtained easily but was interrupted by a local intracranial relapse 57 months after diagnosis (37 months after cessation of therapy; at present the patient is still alive and receiving palliative treatment). This report is warranted because of the rarity of this condition in children. PMID- 10845236 TI - Emergency contraceptive pills. PMID- 10845235 TI - Oral contraceptives--an update. AB - Four decades after introduction of the pill, more women than ever are using it. Today's low-dose oral contraceptives are safer and just as effective as earlier pills. Taken regularly, the pill prevents pregnancy almost without fail. Pill users benefit in other ways, too, such as less anemia and protection from certain cancers. Lower doses have reduced the circulatory disease risks of the pill. PMID- 10845237 TI - Intermediate filaments: molecular architecture, assembly, dynamics and polymorphism. PMID- 10845238 TI - Channel-forming colicins: translocation (and other deviant behaviour) associated with colicin Ia channel gating. PMID- 10845240 TI - Compression therapy for venous leg ulcers. AB - Venous (gravitational) leg ulcers are unsightly, sometimes painful and often difficult to heal. They are associated with incompetence of valves in the deep leg veins and venous hypertension. The main approaches in the management of venous leg ulcers have been to reduce the 'back pressure' in the veins by surgical removal of any varicose veins, postural drainage (elevation of the legs when the patient is lying or seated), and use of compression therapy with bandages, hosiery or intermittent pneumatic compression. In this article, we review the efficacy and discuss correct use of compression therapy. PMID- 10845239 TI - Heart failure drugs: what's new? AB - Heart failure is common, causes major disability and often shortens life. In the past, drugs such as diuretics and digoxin formed the mainstay of treatment. More recently, angiotensin-converting enzyme (ACE) inhibitors have become a standard part of management. New developments in the drug treatment of heart failure include the possible addition of beta-blockers or spironolactone to diuretic and ACE inhibitor therapy. Also, angiotensin-II receptor antagonists have been proposed both as an alternative and as additional therapy to ACE inhibitors. Here, we discuss the place of these new approaches in the treatment of patients with heart failure due to left ventricular dysfunction. PMID- 10845241 TI - Doxepin cream for eczema? AB - One of the more disruptive symptoms of eczema is itching. Traditionally, it is treated with emollients, topical corticosteroids or oral antihistamines. A topical form of the antidepressant doxepin (doxepin 5% cream; Xepin-Bioglan) is now marketed for "the relief of pruritus associated with eczema" in adults and children aged over 12 years. Here, we review the efficacy and safety of this product and assess its role in the management of eczema. PMID- 10845242 TI - The IPA categories "pharyngeal" and "epiglottal": laryngoscopic observations of pharyngeal articulations and larynx height. AB - The phonetic problem is to describe accurately the articulatory mechanism, or mechanisms, responsible for the production of a series of sounds that are presently labelled on the chart of the International Phonetic Association as either pharyngeal or epiglottal. The sounds on which these categories are based are found in the Semitic languages, in the languages of the Caucasus, and in the languages of the Pacific Northwest coast of North America. In order to reconcile a variety of descriptive terms with a logical phonetic taxonomy, auditorily distinguishable parameters are deduced from a naturally occurring variety of sounds, isolated articulatorily, and observed with a fibreoptic laryngoscope to define a cardinal set of articulatory possibilities. Auditory comparisons with database illustrations of the sounds of various languages inform the production of cardinal values in the laryngoscopic study. Voiceless pharyngeals (fricatives) are identified by aryepiglottic fold constriction and a medial aperture. Voiced pharyngeals (approximants) are identified by aryepiglottic fold constriction and a covered glottis. Trilling can occur laterally along the aryepiglottic folds in either voiceless (fricative) or voiced (approximant) mode. A pharyngeal plosive is identified by full occlusion of the aryepiglottic sphincter. "Epiglottal" sounds, which have been described auditorily as "deeper" or "more extreme" than pharyngeals, are associated with either the trilled varieties of the simple fricative or approximant, or the default raised larynx posture of the aryepiglottic sphincter, with radical retraction of the tongue. They are therefore more severely constricted, but not physiologically "deeper" than simple [symbol: see text] or [symbol: see text]. Pharyngeal articulations may also be produced with larynx lowering. Voiceless pharyngeal [symbol: see text] may be accompanied by lowering of the larynx to distinguish it from a larynx-raised [H] variant. Larynx height variations are also found in "tense/lax" register distinctions. PMID- 10845243 TI - Linguistic models of F0 use, physiological models of F0 control, and the issue of "mean response time". AB - This paper evaluates "mean response time" (MRT), a method used in previous studies to relate physiological evidence (recordings of electromyographic activity in the cricothyroid and sternohyoid) to acoustic evidence (fundamental frequency). Rather than averaging over tokens before correlating these signals, we calculated the best response time (RT) for each token, and evaluated the pattern of variability across utterances. Furthermore, rather than correlating over whole utterances, we correlated electromyographic activity (EMG) to fundamental frequency (F0) only over intervals defined in terms of linguistically significant events in the F0 trace, identified using a linguistically motivated model of English intonation. Steep changes in the F0 tended to have better correlation coefficients than shallow ones, which we relate to the physiological model by noting the complex of components contributing to both signal types. Also, the distribution of lead times was easier to interpret when the two tones delimiting the analysis domain had some tight temporal relationship specified by the intonational phonology. Finally, lead times tended to vary as a function of what preceded the target rise or fall. In short, averaging over signals before analysis obscures patterns of variation in the data which may lead to new insights and to new directions for research. PMID- 10845244 TI - Role of vertical larynx movement and cervical lordosis in F0 control. AB - The role of vertical larynx movement in vocal frequency (F0) change has attracted the attention of many researchers. Recently, Hirai, Honda, Fujimoto, and Shimada (1994) proposed a mechanism of F0 control by vertical larynx movement based on the measurement of magnetic resonance images (MRI). In F0 changes, the larynx moves vertically along the cervical spine, which displays anterior convexity (lordosis) at the level of the larynx. Therefore, the vertical larynx movement results in the rotation of the cricoid cartilage and vocal fold tension changes. The present study reexamines the above mechanism based on a qualitative analysis of midsagittal MRI data using three male subjects with evident cervical lordosis. Tracings of the jaw, hyoid bone, laryngeal cartilage, and cervical spine were compared in high and low F0 ranges. In the high F0 range, the hyoid bone moved horizontally while the larynx height remained relatively constant. In the low F0 range, the entire larynx moved vertically, and the cricoid cartilage rotated along the cervical lordosis. These results indicate that the vertical movement of the larynx comprises an effective F0 lowering mechanism, and suggest that the human morphologies of low larynx position and spinal curvature contribute to voluntary use of the vocal function. PMID- 10845245 TI - Stress-induced variation in F2 trajectories as evidence for coproduction in CV syllables. AB - This study explored the extent to which a model of the acoustic consequences of overlapping, sliding consonantal and vocalic gestures was used to account for stress-induced changes in F2 trajectories occurring in test words embedded in a carrier phrase. Three stress conditions were studied including contrastive stress on test words (CS), contrastive stress on the content word preceding test words (U-CS), and non contrastive stress on test words (NS). F2 onset frequency was used to quantify the extent to which adjacent consonantal and vocalic gestures in stop consonant + vowel syllables were coproduced (i.e., overlapped) in the different stress conditions. The predicted relationship between F2 onset frequency and temporal variation in trajectories was also examined within and across stress conditions. In addition, the effects of stress-induced variation in articulatory scaling on F2 onset frequency were studied and factored into the interpretation of the results. The results indicated that F2 onset frequencies tended to differ for stress conditions characterized by large differences in prominence. Regression analyses predicting temporal variation in trajectories from F2 onset frequency accounted for part of the variance within and across stress conditions. Taken together, the results suggested that a model of overlapping, sliding gestures accounts for only some of the stress-induced variability in F2 trajectories in the current study. PMID- 10845246 TI - The impact of genomics on drug discovery. PMID- 10845247 TI - CCK-B antagonists in the control of anxiety and gastric acid secretion. PMID- 10845248 TI - Application of high-throughput screening techniques to drug discovery. PMID- 10845249 TI - Development of neurosteroid-based novel psychotropic drugs. PMID- 10845250 TI - Benzo[b]pyranols and related novel antiepileptic agents. PMID- 10845251 TI - The opioid growth factor, [Met5]-enkephalin, inhibits DNA synthesis during recornification of mouse tail skin. AB - Opioid peptides serve as tonically active negative growth regulators in renewing and regenerating epithelia. To examine the involvement of opioids in renewal of the stratum corneum after tape stripping of tail skin, C57BL/6 J mice were given systemic injections of the potent opioid antagonist, naltrexone (NTX, 20 mg/kg i.p.) following injury. Blockade of opioid-receptor interaction by NTX for 4 h resulted in an elevation of 36-66% in basal cell DNA synthesis measured 24 h after injury. Injection of the endogenous opioid peptide, [Met5]-enkephalin (OGF, 10 mg/kg i.p.) 4 h before termination, suppressed radiolabelled thymidine incorporation in the basal cell layer by 37-46% at 24 h after wounding. The magnitude of the effects on DNA synthesis of OGF, but not NTX, depended on the timing of administration with respect to injury. OGF maximally depressed basal cell labelling (72%) when given 16 h after tape stripping. Concomitant administration of naloxone (10 mg/kg) with OGF blocked the inhibition of DNA synthesis; naloxone alone at the dosage utilized had no effect on cell labelling. Both OGF and its receptor, OGFr, were detected by immunocytochemistry in the basal and suprabasal cell layers, but not the cornified layer of tape stripped and uninjured tail skin. These results indicate: (a) a native opioid peptide and its receptor are expressed in epidermal cells of injured and uninjured mouse tail skin; (b) removal of the stratum corneum by tape stripping does not disrupt the function of the endogenous opioid growth system; (c) the proliferative response to wounding of the tail is tonically inhibited by the receptor-mediated action of an endogenous opioid peptide; and (d) DNA synthesis by basal cells can be elevated by disrupting opioid peptide receptor interactions. PMID- 10845252 TI - Comparison between different cell kinetic variables in human breast cancer. AB - Cell kinetics holds a prominent role among biological factors in predicting clinical outcome and response to treatment in neoplastic patients. Different cell kinetic variables are often considered as valid alternatives to each other, but the limited size of case series analysed in several studies and the lack of simultaneous determinations of all the variables on the same tumours do not justify this conclusion. In the present study, the correlation between [3H]thymidine labelling index ([3H]dT LI), flow cytometric S phase cell fraction (FCM-S) and Ki-67 immunoreactivity (Ki-67/MIB-1) was verified and the type of correlation with the most important clinical, pathological and biological patient and tumour characteristics was investigated in a very large series of breast cancer patients. Ki-67/MIB-1, FCM-S and [3H]dT LI were determined in 609, 526 and 485 patients, respectively, and all three cell proliferation indices were evaluated in parallel on the same tumour in a series of 330 breast cancer patients. All the cell kinetic determinations were performed within the context of National Quality Control Programmes. Very poor correlation coefficients (ranging from 0.37 to 0.18) were observed between the different cell kinetic variables determined in parallel on the same series of breast cancers. Moreover, Ki-67/MIB-1 and FCM-S showed a significant relationship with histological type, grade and tumour size, whereas statistically significant correlations were not observed for [3H]dT LI. In conclusion, the results show that the different cell kinetic variables provide different biological information and cannot be considered as alternatives to each other. PMID- 10845253 TI - An endogenous melanocyte-inhibiting tripeptide pyroGlu-Phe-GlyNH2 delays in vivo growth of monoclonal experimental melanoma. AB - The melanocyte-inhibiting tripeptide (MTP) pyroGlu-Phe-GlyNH2 is present in tissue cultures of non-transformed melanocytes and melanoma cells and influences melanocyte growth in vitro. The objective of the present study was to investigate a possible effect of MTP on the in vivo growth of B16A2, a monoclonal experimental melanoma. The B16A2 clone was established by the limited dilution technique. It has a reduced DNA content and displays slower growth both in vivo and in vitro compared to the parent cell line (B16). B16A2 cells were injected subcutaneously into hairless mice at four sites (300 000 cells in 0.25 ml buffer/site). MTP was given by i.p. injection 3 times a week at two concentrations (1 pmol and 1 nmol/animal). The control animals received the equal volume of solvent. The animals were sacrificed 1 and 2 weeks after tumour transplantation, and all tumours were weighed. One week after transplantation, the animals who received 1 pmol MTP had fewer tumours and a reduced tumour load. Two weeks after the transplantation, the differences between control and treated animals were no longer observed. The results indicate that MTP temporarily delays in vivo tumour growth. PMID- 10845254 TI - Myosin heavy chain degradation during apoptosis in endothelial cells. AB - The cytoskeleton undergoes dramatic changes during apoptosis and many cytoskeletal proteins are known to be degraded during this process. The number of proteases found to be involved in apoptosis is growing but the role of the proteolysis they cause remains poorly understood. This report describes for the first time that myosin heavy chain is cleaved in aortic endothelial cell apoptosis induced either by tumour necrosis factor-alpha or okadaic acid. The cleavage was specific since a well-defined major 97 kDa fragment of myosin heavy chain was produced. The intermediate filament component vimentin was also cleaved into well-defined fragments (31, 28 and 23 kDa). Kinetic studies showed that proteolysis occurred concomitantly with the morphological changes associated with apoptosis, i.e. cellular condensation and fragmentation in apoptotic bodies. These data suggest that the degradation of myosin and vimentin could be involved in the execution of the morphological alterations observed during apoptotic cell death. PMID- 10845255 TI - A subset of cells expressing SV40 large T antigen contain elevated p53 levels and have an altered cell cycle phenotype. AB - Cells transformed by the simian virus 40 (SV40) large T antigen (Tag) contain elevated levels of cellular p53 protein. To quantify this relationship, levels of p53 were measured in NIH 3T3 cells that expressed different concentrations of Tag. Using immunoblotting, average p53 levels were shown to increase linearly with Tag concentrations in these cell lines. Single-cell measurements were also performed using flow cytometry to measure p53 immunofluorescence. Surprisingly, the flow cytometry experiments showed that two distinct cell populations, based on p53 content, were present in cells expressing high levels of Tag. One cell population contained elevated p53 levels. A second population did not contain elevated p53, even though high concentrations of Tag were present in the cells. This latter cell population did not appear to arise because of mutations in either Tag or p53. The two cell populations also had phenotypic differences. In exponentially growing cells, Tag alters the cell cycle distribution (decreases the percentage of G1 phase cells and increases the percentages of S and G2 + M phase cells). This phenotype was maximum in the cell population containing elevated p53. A lesser phenotype was found in the cell population that did not contain elevated p53. These data show, firstly, that cells can express significant levels of Tag and not contain elevated levels of p53 and, secondly, that elevated p53 correlates with the altered cell cycle distribution produced by Tag in growing cells. PMID- 10845256 TI - Epidemiological characteristics of dogs with Lyme borreliosis in the province of Soria (Spain). AB - From the point of view of the human disease, dogs are the most important animal reservoir of Lyme borreliosis; therefore, they are used as 'sentinel animals'. In order to know the epidemiological characteristics of dogs with antibodies against Borrelia burgdorferi, 146 canine sera samples collected during 1993-94 have been studied. The antibody detection was made by an indirect immunofluorescence method and confirmed by a haemagglutination test. Seventeen dogs (11.6%) were seropositive, of which seven were hunting dogs, three were shepherd dogs, five were domestic pets, one was a watchdog and one was a stray dog. Seven dogs had longhair phenotype, 2 medium length type, 4 hard-hair and the other 4 were short haired. Twelve seropositive dogs were males and five were females. All seropositive dogs were more than 1 year old. The dogs with greater seroprevalence were those that harboured ticks more often than the rest. Spring and summer were the seasons when more ticks were observed. The seroprevalence found in dogs was similar to that previously obtained in humans in this area. This finding gave evidence to an intimate association between human and canine seroprevalence. PMID- 10845257 TI - Limited role of rodents as reservoirs of Borrelia burgdorferi sensu lato in Ireland. AB - Despite the presence of Borrelia burgdorferi s.l.-infected Ixodes ricinus ticks in a west of Ireland location, very few small rodents from the same habitats were infected. Most of the infected ticks contained B. garinii or B. valaisiana, which implicates birds as the important reservoir hosts in this location. PMID- 10845258 TI - Occurrence of Borrelia burgdorferi sensu stricto, Borrelia garinii and Borrelia afzelii in the Ixodes ricinus ticks from Eastern Slovakia. AB - A total of 2816 unfed adults nymphs of Ixodes ricinus ticks were collected from vegetation in Kosice (Eastern Slovakia) from 1994 to 1997. Prevalence of Borrelia burgdorferi s. l. in I. ricinus ticks, detected by dark field microscopy, varied and depended upon the year and the habitat of the collected ticks. The lowest prevalence was observed in 1994 (4.8%). During 1995 it increased to 17.2% and during the next two years decreased to 15.5% and 14.2%. The rate of infection varied from 2.1 to 23.3% within 10 examined habitats of the Kosice area. A different value of relative density of ticks was observed in various habitats. It ranged from 9-212 ticks per collecting hour within one flagged area (600 m2) which is 1.5-35.5 ticks per 100 m2. Eight isolates were obtained from the infected ticks. Electrophoresis and immunoblotting with 6 monoclonal antibodies were used for the identification of Borrelia strains. Three tick isolates were identified as B. burgdorferi s. s. and the other three isolates were found to be B. garinii. One strain reacted as a mixed culture of B. burgdorferi s. s., and B. garinii. The strain originated from the Vihorlat Mountains habitat and was detected by PCR-SSCP as B. burgdorferi s. s. with a small amount of B. afzelii. The obtained results emphasize the epidemiological importance not only of B. garinii and B. afzelii but also of B. burgdorferi s. s. in Central Europe. PMID- 10845260 TI - HIV1 infection in extra-European students in Turin (Italy): epidemiologic pattern and risk factors. AB - The authors studied a population of 5634 students from various continents to determine possible correlations between risk factors and seroprevalence of HIV infection. In the African sample the seroprevalence rate was 3.5%; the age range from 19 to 30 years was most affected; prevalence differences between the sexes were slight (3.5% for the males vs 3% for the females); the evaluation of the medical history (STDs, malaria, TB and hospital stays) showed no statistically significant association with HIV-1 infection; the central-eastern Africa significantly appeared the most affected. PMID- 10845259 TI - Pseudomonas spp. complications in patients with HIV disease: an eight-year clinical and microbiological survey. AB - Two hundred and twenty-four episodes of Pseudomonas spp. complications that occurred in 179 consecutive patients with HIV infection were retrospectively reviewed. Pseudomonas spp. organisms were responsible for 11.6% of 1933 episodes of non-mycobacterial bacterial diseases (5.4% of 1072 episodes of sepsis), observed over an 8-year period; 20.7% of patients experienced disease relapses (45 episodes). These complications mostly involved lower airways (66 cases), urinary tract (53 episodes), and blood (34 cases), with Pseudomonas aeruginosa isolated in 161 episodes, and other Pseudomonas spp. in the remaining 63 cases. An advanced HIV disease was frequently present (as expressed by a prior diagnosis of AIDS, a low CD4+ lymphocyte count, and leukopenia-neutropenia). Indwelling intravascular and urinary catheters were often associated with bacteremia and urinary tract involvement, respectively. More than 60% of patients were given antibiotics and/or cotrimoxazole in the month preceding the onset of Pseudomonas spp. disease. Bacterial strains isolated from our HIV-infected patients showed a favorable sensitivity to piperacillin, ceftazidime, imipenem, amikacin, tobramycin, and ciprofloxacin. An adequate antimicrobial treatment led to clinical and microbiological cure in 73.2% of patients at the first episode, and in 22.3% more subjects after one or more relapses. A lethal outcome occurred in only eight patients of 179 (4.5%), suffering from a far advanced HIV disease; P. aeruginosa infection directly contributed to death in four cases (sepsis, and/or pneumonia). Nosocomial disease occurred in 46.4% of the 224 episodes, and was significantly related to a previous diagnosis of AIDS, concurrent neutropenia, the occurrence of sepsis or urinary tract infection, disease relapses, the involvement of non-aeruginosa Pseudomonas spp., and a lethal outcome, compared with community-acquired infection. Our experience (the largest reported to date) confirms that Pseudomonas spp. (including non-aeruginosa Pseudomonas spp. organisms) is responsible for remarkable morbidity and mortality among patients with HIV infection, and may pose relevant problems to clinicians and microbiologists involved in the care of HIV-infected patients. PMID- 10845261 TI - HIV testing and retesting for men and women in Switzerland. AB - This study was conducted to describe voluntary HIV testing in the general population in Switzerland and to estimate yearly HIV test incidence. In 1994, a representative telephone survey of individuals aged 17 to 45 years obtained self reported information on HIV testing. In addition to describing cumulative HIV test incidence, yearly HIV test incidence over time was estimated by a Bayesian hurdle model allowing for the plausible scenario of test consumption differing between first test and subsequent retests. Overall, 33% of the Swiss population (age 17 to 45 years) has been tested at some time for HIV on a voluntary basis (30% men, 36% women). For the time period 1990-1994, the result showed for 35 year-old individuals with supposedly low risk behavior, that 1) annual test incidence (first test or retest) showed a greater increase for men (4.2 to 5.9%) than for women (5.0 to 6.0%); 2) annual first test incidence increased moderately and differed for men and women (2.9 to 3.4% for men, 4.6 to 5.2% for women), and 3) annual retest incidence was twice as high for men (17.6%) as for women (8.6%). In conclusion, a substantial part of the Swiss population has been tested at some stage for HIV on a voluntary basis, and differences exist for testing and retesting between men and women. PMID- 10845262 TI - Surveillance of multidrug-resistant strains of Salmonella enterica serotype Typhimurium in southern Italy in the years 1992-1997. AB - Spread of multidrug-resistant strains of Salmonella enterica serotype Typhimurium (S. typhimurium) is increasingly reported worldwide. The presence of a pattern of resistance to ampicillin, chloramphenicol, streptomycin, sulfonamides and tetracycline (ACSSuT), in some cases associated to trimethoprim and infrequently to quinolones, is of particular concern. This resistance pattern appears to be chromosomally encoded and, in most epidemiological studies, closely related to definitive type 104 (DT104). In southern Italy multidrug-resistant isolates of S. typhimurium had been identified since 1980, but only during 1992 S. typhimurium strains with chromosomally encoded drug resistance were first isolated from domestic animals. One hundred fifty-five isolates--52.5% of the multidrug resistant strains identified in the years 1992-1997--were submitted to phage typing and plasmid profile analysis. Ribotyping was also performed in comparison with a random sample of 150 strains susceptible or resistant to three or less antibiotics identified in the same interval of time. Four ribotypes (RTs)--1, 5, 8, and 48--included approximately 90% of the multiresistant strains, RT8 accounting for 61.2%. Phage type (PT) 193 is the most prevalent phage type. Phage typing and ribotyping suggest that few bacterial clones are involved in spread of multidrug-resistant S. typhimurium strains in southern Italy. PMID- 10845263 TI - Salmonella in slaughter pigs of German origin: an epidemiological study. AB - The Salmonella prevalence in slaughter pigs of German origin was determined in seven abattoirs located in different regions of the country between February and June 1996. A total of 11,942 pigs delivered to the abattoirs in 752 batches, most of them comprised of pigs from individual finishing farms, was investigated by the bacteriological examination of faecal and gut lymph node samples, as well as of surface swabs taken from the carcasses. Salmonellae were isolated from 3.7% of the faecal samples, 3.3% of the lymph nodes and 4.7% of the surface swabs. The estimated overall prevalence of Salmonellae was 6.2% in the slaughter pigs, ranging between 1.9% and 12% in individual abattoirs. In the samples taken from carcasses, the estimated prevalence of Salmonellae reached 10.3%. 648 out of 752 batches could be included in a statistical analysis. No Salmonellae were detected in nearly 70 percent of the batches included in this analysis (n = 648). High Salmonella prevalences of more than 50 percent positive animals were detected only in 13 batches (2.0%). A statistically significant influence of the duration of the transport of slaughter pigs to the abattoirs or the waiting period in the abattoirs prior to slaughter could not be detected. PMID- 10845264 TI - Capture-recapture method to determine the best design of a surveillance system. Application to a thyroid cancer registry. AB - The capture-recapture method is often confronted, when assessing completeness of surveys, to problems of dependence of data sources. The objective of this paper is to discuss the application of capture-recapture methods to choose the optimal combination of sources for a surveillance system. Our approach is based on: (1) using multiple sources, (2) assessing dependence between sources and between pools of dependent sources and other sources, (3) ruling out combinations that yield biased estimates, and (4) choosing the combinations of sources that have the best ratio between precision and applicability. We studied the independence for each pair of sources by computing the capture-recapture odds ratio. We characterized all combinations of sources by their sensitivity, coefficient of variation of the estimated number of cases, and level of resources needed to ascertain cases. Application of the approach is illustrated by data from a survey of thyroid cancer in New Caledonia, where five sources were used to estimate the incidence. The five sources provided 119 cases; the exhaustivity of sources and combinations of sources varied from 27.1 to 99.2%. Determination of dependence revealed ten dependencies out of 22 combinations. Coefficients of variation of the estimated number of cases varied from 0.83 to 27.79. The preferred combination included four sources and had a sensitivity of 97.5 and a coefficient of variation of 0.94. An assessment of dependence, based on simple criteria, can be used to choose the best combination of sources for a registry or a surveillance system. PMID- 10845265 TI - Use of unconventional methods of therapy by cancer patients in Pakistan. AB - Most of the studies related to the use of unconventional methods of therapy by cancer patients have been carried out in the developed countries. This study was conducted to ascertain the frequency, type, and duration of use of unconventional methods of therapy by cancer patients in Pakistan. We also wanted to identify individuals who are most likely to use these methods and to compare the findings with those reported from the developed countries. Between 1 April and 30 May 1994, all patients with histologically proven cancer who visited the oncology unit were interviewed. A printed questionnaire with questions and options was used as an interview guide. Informed consent was obtained. One hundred and ninety one patients were interviewed, on average, for 25 minutes each. Use of unconventional methods of therapy by cancer patients was widespread (54.5% of all patients). The majority (83.7%) were influenced by family members to use these methods. Traditional herbal medicines (70.2%) and homeopathy (64.4%) were the most commonly employed methods. Thirty-six percent of the users employed these methods before receiving any conventional therapy. Only 15% used these methods after conventional therapeutic options had been exhausted. Patients generally perceived these methods as useful, non-toxic and inexpensive. Age, marital status, socio-economic background, education level and status of underlying neoplasm did not influence the frequency of use of unconventional methods. The use, however, was influenced by gender, family size, and type of underlying malignancy. Patients aware of their diagnosis were less likely to use these methods. This study suggests that use of unconventional methods by cancer patients in Pakistan is widespread. Unlike western countries, these methods are often employed before receiving any conventional therapy. This probably results in a significant delay which can be expected to adversely influence the subsequent disease management and survival. Public education, reduction of cost and easy availability of conventional therapy may be helpful in reducing the use of methods which otherwise may have no proven value. PMID- 10845266 TI - Life expectancy as an indicator of environmental health. AB - Whether or not life expectancy at birth is related to the quality of life as expressed by global economical, environmental and nutritional measures is the primarily studied question in this article. Two models, set of independent variables and multivariate analysis was performed. An attempt to estimate the role of studied variables in overall life expectancy was done, too. A descriptive, ecological study design was used. The population of 156 countries have been taken into account, using data from published databases [7, 9]. Access to safe drinking water, per capita gross domestic product, literacy, calories available as percentage of needs and per capita public health expenditures were taken as exposure, and compared with life expectancy at birth. A linear regression model was used to estimate the role of different exposures on life expectancy at birth. A correlation matrix for all variables and life expectancy at birth is presented in the article. Literacy and access to safe drinking water are statistically significant variables (p < 0.001) also after fitting a linear regression model. The correlation coefficient for the linear model was 0.8823 (R2 = 0.7784). Shares of years of life from overall life expectancy attributed to studied variables were 28.06, 9.42, 2.04 and 1.93% for literacy, access to safe drinking water, GDP and calories available as percentage of needs, respectively. PMID- 10845267 TI - Asthma during pregnancy--a population based study. AB - To study delivery outcome in women with asthma, using Swedish health registers. Women with asthma were identified in two ways: by information in interviews performed by midwives at the pregnant woman's first visit to antenatal care, and by linkage between a medical birth register and a hospital discharge register, identifying women who had been hospitalized for asthma and also had a delivery. Births between 1984 and 1995 were studied. An increased risk for preterm birth and low birth weight was seen, possibly co-varying with disease severity. Also a significant increase in pregnancies of more than 41 weeks duration was noticed. An increase in infant death but not in congenital malformations rate was observed. An association with preeclampsia, gestational diabetes, and infant hypoglycemia was verified. Maternal asthma appears to be a risk factor for preterm and postterm births and increases the risk for some pregnancy complications. PMID- 10845268 TI - The economic burden of hepatitis B in Germany. AB - Despite the widespread distribution of Hepatitis B virus (HBV) infection and the ongoing controversy about HBV immunization, surprisingly few published studies examined in detail the economic impact of HBV infection in Europe. Therefore, we investigated a cohort of 180 patients throughout Germany to evaluate the economic burden of HBV-associated disease. In 58 patients with acute and 122 patients with chronic HBV infection, cost-consuming events including direct medical costs and work-loss costs were documented. The direct costs were DM 7702 (95% confidence interval (CI): 5473-9931) for each acute HBV infection and DM 4247 (CI: 1601 6893) per patient-year of chronic HBV infection, with marked differences between different stages of HBV disease. The derived overall costs (1997 price levels) per year were DM 10,018 (CI: 7613-12,421) and DM 4860 (CI: 2185-7536), respectively. Based on crude population-based estimates (30,000 acute and 420,000 chronic HBV cases), we calculated the total HBV-related costs in Germany to exceed DM 1200 millions in 1997 (CI: 924.2-1536.7), with the treatment of patients with chronic active HBV disease as the major cost determinant. Previously published data from Germany probably overestimated the financial impact of acute HBV infection. In summary, our results illustrate the ongoing economic importance of this potentially life-threatening, but preventable disease and support the call for more accurate HBV surveillance and control in Germany. PMID- 10845269 TI - A retrospective study of autochthonous strongyloidiasis in Region Midi-Pyrenees (Southwestern France). AB - In order to assess the existence of autochthonous strongyloidiasis, a retrospective study was carried out in Region Midi-Pyrenees (Southwestern France). Among 63 strongyloidiasis cases diagnosed at the Consultation of the Department of Parasitology, CHU Purpan in Toulouse, 17 patients (27%) were identified as likely harboring an autochthonous infection. The diagnosis was based upon the results of either Baermann's method for stool examination, or indirect fluoroimmunoassay using Strongyloidiasis ratti L3 larvae as antigen. Repeated contact with soil or mud, due to occupation or gardening, was found in 13 patients. Twelve cases lived in the upper basin of the Garonne River. The clinical and laboratory findings were similar to those reported by the literature. No case of larva currens was observed. These results suggest that a permanent transmission of strongyloidiasis possibly exists in Region Midi Pyrenees, requiring further prospective studies. PMID- 10845270 TI - Dietary assessment of an educated young Spanish population using a self administered meal-based food frequency questionnaire. AB - In this study a self-administered meal-based semi-quantitative food frequency questionnaire was developed, whose main aim was to classify individuals by their intake of food groups and nutrients. The respondents (205 final participants) from the target population were recruited from students and staff of the Faculty of Chemistry of the University of Castilla-La Mancha in Ciudad Real (Spain). The validity of the questionnaire was assessed in comparison with a four-day weighed diet record on a subset group of participants (n = 38). Results of the calibration study were similar to those reported in the literature, showing a reasonable correlation between the two methods. To improve dietary habits of the target population, an effective nutritional education programme should stress the importance of increasing consumption of food groups rich in starch and reducing those containing high level of proteins, sugars, fat and cholesterol. PMID- 10845271 TI - Spontaneous pneumothorax related with climatic characteristics in the Valencia area (Spain). AB - Spontaneous pneumothorax (SP) is a common respiratory condition that is easily recognized and treated, though the etiology remains largely unknown. Apart from individual factors, it has been suggested that changes in atmospheric pressure may trigger the onset of SP. Our aim was to examine the association between the onset of episodes of SP with atmospheric pressure changes in the general population of an area with a Mediterranean climate. A prospective survey was conducted to detect all episodes of SP in a region of Valencia (Spain) from January 1994 to January 1996. A total of 69 cases were identified; of these, 62 episodes of SP met the inclusion criteria for this survey. Climatic measurements during the two years studied were obtained from the Spanish National Meteorological Institute, including 6-hour readings of temperature, relative humidity, clear sunlight, atmospheric pressure, visibility, wind speed and rainfall. Associations were evaluated between the daily number of hospital admissions for SP and diurnal changes in atmospheric pressure involving rises and falls of as much as 7 and 10 mbar. In addition, the association of atmospheric pressure changes to time periods of 1 a.m., 7 a.m., 1 p.m. and 7 p.m. was assessed. Drops in atmospheric pressure below the fifth percentile and elevations above the ninety-fifth percentile were regarded as 'unusual'. There was no association between the development of SP and changes in atmospheric pressure, and no correlation was found with repeated exposure to unusual changes of atmospheric pressure. Further research is needed using similar methods to establish comparisons between countries with different climates. PMID- 10845272 TI - Polyamine profiles in tumor, normal tissue of the homologous breast, blood, and urine of breast cancer sufferers. AB - Polyamines are involved in the development of breast cancer. We assayed polyamines in erythrocytes, urines, and breast tissues (tumor tissue and histologically normal breast tissue close to the tumor) of patients with invasive breast cancer (n = 174) and benign breast disease (n = 71, used as controls). Polyamine levels in red blood cells and urine were similar to the polyamine concentrations found in healthy subjects, and thus cannot be used as diagnostic markers of breast cancer. In cancer tissue, polyamines were significantly increased in comparison with the polyamine concentrations in controls, and were correlated to the tumor aggressiveness as evaluated by histological grade and Ki 67 proliferative index. On the other hand, correlation was found between polyamine levels in the tumor and the status of the hormone receptors. In the mammary tissue close to the cancer, polyamines dramatically decreased in comparison with the polyamine levels of tissue samples removed around the histologically proven benign tumors. The changes of the polyamine concentrations in the histologically normal breast tissue in the vicinity of the cancer could play a role in the cancer development and need further studies, especially if polyamines are considered as a potential therapeutic target in breast cancer. PMID- 10845273 TI - T cell suppression as a mechanism for tolerance to MUC1 antigen in MUC1 transgenic mice. AB - C57BL/6 mice transgenic for human MUC1 (MUC1.Tg) have been developed to study immunologic responsiveness to the human MUC1 tumor-associated antigen. In the present studies, MUC1.Tg mice were immunized with purified human MUC1 antigen and irradiated MUC1-positive (MC38/MUC1) tumor cells. Immunization with MUC1 antigen was associated with induction of an anti-MUC1 antibody response and no detectable cytotoxic T cell reactivity. Similar findings were obtained after immunization with irradiated MC38/MUC1 tumor cells. The results also demonstrate that immunization of MUC1.Tg mice with MUC1 is associated with decreased levels of CD8+ T cells. In addition, expression of alphabeta T cell receptors and CD28 were down-regulated on CD8+ T cells as a consequence of MUC1 immunization. These findings support a role for T cell suppression in tolerance to MUC1 antigen in MUC1.Tg mice. PMID- 10845274 TI - Cellular responses of mammary carcinomas to aromatase inhibitors: effects of vorozole. AB - Vorozole (Vz) is a competitive non-steroidal inhibitor of aromatase, which has been used to treat breast cancer in postmenopausal women and in various chemoprevention pre-clinical studies. Recently, we assessed the inhibitory effect of Vz on MNU-induced mammary carcinogenesis (Lubet et al., 1994), as well as on the progression of mammary tumors (Lubet et al., 1998). In this study we evaluated the effects of Vz on tumor growth, serum estradiol, cell proliferation, apoptotic and non-apoptotic cell death to determine whether any of these 'surrogate' markers might reflect the efficacy of various doses of Vz. Vz at doses of 2.5 (Hi), 0.32 (Md), and 0.08 (Lo) mg/kg body weight induced complete (100%), 60%, and 20% regression of mammary tumors, respectively. Vz at Hi and Md doses caused a decrease in serum estradiol within the first two days of treatment, and the estradiol values remained low with additional treatment for 4 and 10 days. When Vz was administered to animals bearing palpable tumors a time and dose-dependent decrease in the proliferating cells (BrdU-L1) was observed. The percentage of apoptotic cells (A1) sharply increased 2 days after initiation of Vz treatment and then decreased followed by an increase in non-apoptotic dead cells. Interestingly even the Lo dose of Vz, which was only moderately effective in suppressing tumor growth, decreased cell proliferation and increased cell death in the peripheral tumor areas at 4 and 10 days after initiation of treatment. The time- and dose-dependent alterations in various cell parameters suggest two different phases of Vz-induced cellular responses: (1) an early phase (2-4 days of treatment) with a sharp increase in apoptotic cells and decrease in proliferating cells, and (2) a later phase (10 days) with disintegration of tumor parenchyma, increase in non-apoptotic dead cells, and decrease in apoptotic cells. The dose-dependent decrease in proliferating cells and increase in apoptotic and non-apoptotic cell death in Vz-treated animals suggest that these biomarkers might be used as potential surrogate endpoints for efficacy in breast cancer chemoprevention and therapy studies with aromatase inhibitors. PMID- 10845275 TI - Anti-tumor immunity induced by interleukin-12 gene therapy in a metastatic model of breast cancer is mediated by natural killer cells. AB - An intrahepatic tumor model for metastatic breast cancer was generated in syngeneic mice by direct inoculation of JC cells, a murine mammary adenocarcinoma cell line. Intratumoral administration of a recombinant adenoviral vector expressing the murine Interleukin-12 (ADV-mIL-12) resulted in significant reduction in the tumor volume compared to control vector. Tumor regression was also evident on histopathologic analysis of the liver, where inflammatory changes as opposed to nuclear atypia predominated after IL-12 vector treatment. There was a significant prolongation in the long term survival of IL-12 treated animals, with complete tumor rejection in 40% of the animals. In vivo depletion studies using specific monoclonal antibodies against the various lymphocyte subsets showed a significant reduction in long term survival after natural killer (NK) cell depletion. This suggests that the NK cell is a critical effector in the antitumor effect mediated by IL-12. These results strongly support the potential role of gene mediated cytokine therapy for the treatment of metastatic breast cancer. PMID- 10845276 TI - Microsatellite instability markers in breast cancer: a review and study showing MSI was not detected at 'BAT 25' and 'BAT 26' microsatellite markers in early onset breast cancer. AB - Microsatellite markers may provide evidence of faulty DNA mismatch repair (MMR) via the detection of microsatellite instability (MSI). The choice of microsatellite markers may impact on the MSI detection rate. In hereditary non polyposis colon cancer (HNPCC), several informative microsatellite markers have been recommended. Two of these, BAT 25 and BAT 26, are quasi-homozygous, enabling analysis of tumour DNA in the absence of paired normal DNA. Sixty-six breast cancer patients under 45 years of age at diagnosis were examined for MSI at BAT 25 and BAT 26. Tumour DNA was extracted from paraffin-embedded tissue. No MSI was detected at the BAT 25 or BAT 26 loci. An additional five microsatellite markers, known to be informative for HNPCC, were examined for MSI in these patients. Apparently-normal profiles were achieved. A tabulated survey of 306 microsatellite markers used to detect MSI in breast cancer revealed that only 35.5% of markers detected MSI at an average rate of 2.9%. The MSI detection rate at the specific HNPCC markers varied from 0% to 10% in breast cancer, with D175250 and TP53 being the HNPCC markers most suitable for analysis of breast cancer. The size of the microsatellite marker's repeat unit did not impact on MSI detection rates. Compiled data from large studies (n > 100) revealed D115988 as the marker with the highest MSI detection rate. Genomic instability pathways of carcinogenesis, characterised by MMR defects and MSI, appear to play a role in the genesis of some breast cancer types. PMID- 10845277 TI - RT-PCR amplification of CK19 mRNA in the blood of breast cancer patients: correlation with established prognostic parameters. AB - We optimized the assay for detection of cytokeratin 19 (CK19) mRNA by the reverse transcriptase-polymerase chain reaction (RT-PCR) in blood as an index of circulating tumor cells in breast cancer patients. The limit of detection of < 1 MCF7 tumor cells per 10(6) peripheral blood leukocytes (PBL) was achieved in mixing experiments. We did not detect CK19 mRNA in control bloods (0/30) or in the blood of patients with benign breast disease (0/15). In blood samples from 109 patients with invasive breast cancer, CK19 mRNA was detected in 7/23 patients with node-negative disease, in 21/58 with node-positive disease, and in 20/28 with distant metastases. There was a significant association (P < 0.01) of CK19 positivity with distant metastatic versus both node-negative and node-positive disease, but not with any other histopathological parameter examined. In a small number of patients with distant metastases, increased intensity of the CK19 RT PCR signal was associated with a reduced survival. PMID- 10845278 TI - Beta-adrenoceptor signaling and its control of cell replication in MDA-MB-231 human breast cancer cells. AB - MDA-MB-231 human breast cancer cells express high beta-adrenoceptor levels, predominantly the beta2 subtype. Receptor stimulation by isoproterenol evoked immediate reductions in DNA synthesis which were blocked completely by propranolol and were of the same magnitude as effects elicited by high concentrations of 8-Br-cAMP. Isoproterenol-induced inhibition of DNA synthesis was maintained throughout several days of exposure, resulting in a decrement in total cell number, and the effects were augmented by cotreatment with dexamethasone; an even greater effect was seen when cAMP breakdown was inhibited by theophylline, with or without addition of isoproterenol. Despite the persistent effect of isoproterenol, receptor downregulation was evident with as little as 1 h of treatment, and over 90% of the receptors were lost within 24 h. Receptor downregulation was paralleled by homologous desensitization of the adenylyl cyclase response to beta-adrenoceptor stimulation. Dexamethasone augmented the effects of isoproterenol on DNA synthesis but did not prevent receptor downregulation or desensitization. These results indicate that beta adrenoceptors are effectively linked, through cAMP, to the termination of cell replication in MDA-MB-231 human breast cancer cells, and that activation of only a small number of receptors is sufficient for a maximal effect. Novel pharmacologic strategies that focus on cell surface receptors operating through adenylyl cyclase may offer opportunities to combat cancers that are unresponsive to hormonal agents, or that have developed multidrug resistance. PMID- 10845279 TI - Eye problems in breast cancer patients treated with tamoxifen. AB - BACKGROUND: Tamoxifen is an oral anti-estrogen used in the treatment of breast cancer and recently approved to reduce the incidence of breast cancer in high risk women. As a large clinical trial of tamoxifen has reported an increased risk of cataract, we conducted a study of women with breast cancer to evaluate the association of tamoxifen with cataracts and other eye problems. METHODS: We attempted to recruit previously interviewed patients who were cases in a population-based case-control study of 2653 women with primary breast cancer diagnosed between 1987 and 1996 at ages 55-72 years in Los Angeles County, California, USA. In November 1997, each case was mailed a questionnaire to ascertain self-reported incidence of eye diseases and Amsler grid test scores. Information from 1297 women aged 57-75 years of age was analyzed. Women reporting treatment with tamoxifen were categorized as standard-term users (4-5 years), short-term users (<4 years), or long-term users (6+ years) and compared to non users. All p-values, relative risks, and confidence intervals for differences in eye problems and grid test results are adjusted for age and stage of disease at diagnosis. FINDINGS: Standard-term and long-term users of tamoxifen more frequently reported developing cataracts than non-users (18.2%, 21.4% vs. 14.8%). The relative risk was 1.40 (95% confidence interval 0.94-2.10) for standard-term users and 1.70 (1.11-2.59) for long-term users. Tamoxifen use was unrelated to frequency of glaucoma or macular degeneration or to Amsler grid test results. INTERPRETATION: Our study suggests that five or more years of tamoxifen use increases risk of cataracts. Healthy women considering tamoxifen use to reduce risk of breast cancer should be advised of the possibility of cataract development. Women choosing such therapy should be diligent about receiving regular ocular exams. PMID- 10845281 TI - EORTC 10941: A phase II study of liarozole in postmenopausal patients with 'chemotherapy-resistant' or 'potentially hormone sensitive' metastatic breast cancer. AB - Liarozole is an imidazole compound that inhibits enzymes involved in steroid hormone aromatisation and retinoid metabolism. The IDBBC branch of the EORTC has performed a series of phase II studies of the agent in four groups of postmenopausal women with metastatic breast cancer. This paper reports the results of the first two groups: 'Chemotherapy Resistant' (unrestricted ER status, 1 or 2 prior chemotherapy regimens, 0-2 prior hormonal therapies) and 'Potentially Hormone Sensitive' (ER positive or unknown, 1 or 2 prior hormonal therapies with a substantial disease free interval or progression free survival, and no history of chemotherapy for metastatic disease). Liarozole was administered at 150-300 mg orally bid. The objective response rate was 12% in the 'Chemotherapy Resistant' group (n = 34), and 22% in the 'Potentially Hormone Sensitive' group (n = 37), with median response durations of 9 and 14 months, respectively. Median time to treatment failure was only 2 months in both groups, due largely to the significant percentage (24%) of patients who ceased treatment following excessive mucocutaneous and gastrointestinal toxicity. This adverse event profile will limit its use in breast cancer. Results of the 'ER negative' and 'Tamoxifen Refractory' groups will be reported in a future paper. PMID- 10845280 TI - Analysis of cathepsin D in human breast cancer: usefulness of the processed 31 kDa active form of the enzyme as a prognostic indicator in node-negative and node positive patients. AB - The relative amounts of the precursor (52 kDa) and processed (31,27 kDa) forms of cathepsin D have been analyzed by Western blotting in biopsied breast tissue cytosols from 134 lesions from invasive breast cancer patients, 24 lesions from patients with ductal carcinoma in situ (DCIS), 227 lesions from benign breast disease patients, and 28 lesions from normal control subjects. The mean relative percentage amount of the 31 kDa form was significantly increased (p < 0.001) in the invasive breast cancer group compared to the other three groups. In addition, the mean relative percentage amount of the 31 kDa form was significantly increased (p < 0.05) in node-positive compared to node-negative breast cancer patients. In the benign breast disease group, patients with proliferative-type disease had a significantly increased (p = 0.02) mean relative percentage amount of the 31 kDa form of cathepsin D compared to patients with nonproliferative-type disease. Invasive breast cancer patients were followed for up to 75 months to determine if the relative percentage amount of the 31 kDa form of cathepsin D was predictive of disease-free and overall survival. Although the amount of the 31 kDa form was not predictive of disease-free survival, patients in the 'high' 31 kDa group (> 18%) were significantly (p < 0.05) more likely to die than patients in the 'low' 31 kDa group (< or = 18%). The 12 patients who died were all node positive and in the high 31 kDa group. It thus appears that the relative amount of the processed, active 31 kDa form of cathepsin D is a useful prognostic indicator, at least in node-positive breast cancer patients. PMID- 10845282 TI - The angiosomes of the head and neck: anatomic study and clinical applications. AB - The angiosome concept was introduced over a decade ago by Taylor and Palmer, whereby the body was considered to be composed anatomically of multiple three dimensional composite blocks of tissue supplied by particular source arteries. Since then, detailed studies of the forearm and leg have been examined by Taylor and his coworkers. This study focuses on another region--the head and neck. Six fresh head and neck cadaver specimens were examined after infusion with a radio opaque lead oxide mixture and correlated with over 24 previous body studies. The vascular anatomy of the skin, superficial musculoaponeurotic system (SMAS), muscles, brain, dura, and bone was examined. Each layer was painstakingly removed, photographed, labeled, and mapped to the respective arteries and veins. A radiologic subtraction technique was used to allow successive layers to be compared. This information was then scanned into a computer, analyzed, color coded, and labeled, thereby producing a three-dimensional study of the head and neck region to identify the respective angiosomes. As in previous detailed examinations of the leg and forearm, the angiosomes were found to be connected usually within tissues, such as muscle, skin, specialized organs or glands, rather than between the tissues. The muscles usually had vessels of two or more angiosomes supplying them and fell into three major groups based on the similarity of their arterial supply. In some areas, the midline anastomoses were rich, especially in the integument of the scalp, forehead, and lips. In other regions, the midline vascular connections were poor, especially in the tongue and palate. No fewer than 13 angiosomes of the head and neck, supplied by the branches of the external carotid, internal carotid, and subclavian arteries, have been defined, mapping their three-dimensional territories in the skin, the deep soft tissues, and the bones. Although most angiosomes spanned between skin and bone, three territories, those of the vertebral, lingual, and ascending pharyngeal vessels, were confined to the deep tissues without cutaneous representation. Finally, this study provides additional data for the surgeon to help plan safer incisions and better reconstructive flap procedures. It also gives information that may help explain the etiology and treatment of head and neck arteriovenous vascular malformations. PMID- 10845283 TI - Analysis of fronto-orbital advancement for Apert, Crouzon, Pfeiffer, and Saethre Chotzen syndromes. AB - The purposes of this study were (1) to document outcome after primary fronto orbital advancement for the four major eponymous craniosynostotic syndromes (Apert, Crouzon, Pfeiffer, and Saethre-Chotzen) and (2) to identify factors that might influence need for primary and secondary fronto-orbital advancement or foreheadplasty. Also tested was the hypothesis that coincident sagittal synostosis could modulate brachycephaly and affect whether a primary or secondary frontal operation was necessary. Data were collected on age and indications for initial operation, type of primary and secondary frontal procedures, and concomitant sagittal synostosis. Patients initially managed by subcranial Le Fort III were included in the study group but excluded from analysis of fronto-orbital advancement. Patients treated by monobloc advancement or Le Fort III osteotomies with frontal grafting or Anderl modification were assessed as having had primary fronto-orbital advancement. Minimum time to follow-up was 5 years. A total of 126 patients met inclusion criteria. Lateral photographs were examined to assess preoperative and postoperative sagittal position of supraorbital rims-to-globes. Frontal re-advancement was indicated if the corneal apex was anterior to the supraorbital rim. Foreheadplasty was indicated for unacceptable frontal contour and normal supraorbital rim-to-globe relationship. Primary correction for frontal retrusion was not required in 4 percent of Apert (1 of 25), 16 percent of Crouzon (7 of 44), 6 percent of Pfeiffer (2 of 31), and 19 percent of Saethre-Chotzen (5 of 26) patients. Of those infants who had a primary fronto-orbital advancement, reoperation for either supraorbital retrusion or frontal deformity was necessary in all 16 Apert patients and in 5 of 19 Crouzon (26 percent), 10 of 26 Pfeiffer (38 percent), and 13 of 20 Saethre-Chotzen (65 percent) patients (p < 0.001). Age at initial fronto-orbital advancement did not influence reoperative rate. No correlation was found between concomitant sagittal synostosis and necessity for primary or secondary frontal correction (p = 0.22). In summary, phenotypic diagnosis was determinant for outcome as defined by need for secondary fronto orbital advancement, foreheadplasty, or both. Apert patients had the highest incidence of reoperation for frontal retrusion or forehead contour. Crouzon and Saethre-Chotzen patients were most likely to express a minor phenotype and not require fronto-orbital correction. Coincident sagittal synostosis did not influence frontal projection, as reflected in need for either primary or secondary frontal advancement. PMID- 10845284 TI - Distraction osteogenesis for obstructive apneas in patients with congenital craniofacial malformations. AB - Infants with congenital craniofacial malformations often have associated severe mandibular hypoplasia causing obstruction of the hypopharynx by retroposition of the base of the tongue into the posterior pharyngeal airway. Management depends on the severity of the airway obstruction. Most cases can be managed by prone positioning until the infant outgrows the problem at 3 to 6 months of age. In more critical cases, monitoring of oxygen saturation, temporary placement of a nasopharyngeal tube, and placement of an endotracheal tube will be useful procedures. Tracheotomy is an effective method for more severe cases, but longstanding tracheotomies result in high morbidity and occasional mortality. Mandibular distraction was performed in seven patients, ranging in age from 1 to 18 months, with critical obstructive apnea secondary to mandibular hypoplasia characterized by an apnea/hypopnea index greater than 20 apneas per hour and oxygen saturation below 80 percent. Two patients were tracheotomized previously. Mandibular lengthening, from 16 to 25 mm on the left side and from 10 to 22 mm on the right, was achieved in 21 to 25 days. Improvement of airway obstruction parameters was measured on polysomnograms and lateral cephalograms. Mandibular lengthening by gradual distraction is a successful method for young patients with severe mandibular hypoplasia causing critical obstructive apneas. Avoidance of tracheotomy or early decannulation in previously tracheotomized patients is a great advantage for patients with congenital craniofacial malformation. PMID- 10845285 TI - A classification system and algorithm for reconstruction of maxillectomy and midfacial defects. AB - Maxillectomy defects become more complex when critical structures such as the orbit, globe, and cranial base are resected, and reconstruction with distant tissues becomes essential. This study reviews all maxillectomy defects reconstructed immediately using pedicled and free flaps to establish (1) a classification system and (2) an algorithm for reconstruction of these complex problems. Over a 5-year period, 60 flaps were used to reconstruct defects classified as the following: type I, limited maxillectomy (n = 7); type II, subtotal maxillectomy (n = 10); type IIIa, total maxillectomy with preservation of the orbital contents (n = 13); type IIIb, total maxillectomy with orbital exenteration (n = 18); and type IV, orbitomaxillectomy (n = 10). Free flaps (45 rectus abdominis and 10 radial forearm) were used in 55 patients (91.7 percent), and the temporalis muscle was transposed in five elderly patients who were not free-flap candidates. Vascularized (radial forearm osteocutaneous) bone flaps were used in four of the 60 patients (6.7 percent) and nonvascularized bone grafts in 17 (28.3 percent). Simultaneous reconstruction of the oral commissure using an Estandler procedure was performed in 10 patients with maxillectomy and through-and-through soft-tissue defects. Free-flap survival was 100 percent, with reexploration in five of 55 patients (9.1 percent) and partial-flap necrosis in one patient. Seven of the 60 patients (11.7 percent) had systemic complications, and four died within 30 days of hospitalization. Fifty patients had more than 6 months of follow-up with a mean time of 27.7 (+/- 15.6) months. Postoperative radiotherapy was administered in 32 of these patients (64.0 percent). Chewing and speech functions were assessed in 36 patients with type II, IIIa, and IIIb defects. A prosthetic denture was fixed in 15 of 36 patients (41.7 percent). Return to an unrestricted diet was seen in 16 patients (44.4 percent), a soft diet in 17 (47.2 percent), and a liquid diet in three (8.3 percent). Speech was assessed as normal in 14 of 36 patients (38.9 percent), near normal in 15 (41.7 percent), intelligible in six (16.7 percent), and unintelligible in one patient (2.8 percent). Globe and periorbital soft-tissue position was assessed in 14 patients with type I and IIIa defects. There were no cases of enophthalmos, and one patient had a mild vertical dystopia. Ectropion was observed in 10 of 14 patients (71.4 percent). Oral competence was considered good in all 10 patients with excision/reconstruction of the oral commissure; however, two patients (20 percent) developed microstomia after receiving radiotherapy. Aesthetic results were evaluated at least 6 months after reconstruction in 50 patients. They were good to excellent in 29 patients (58 percent) for whom cheek skin and lip were not resected, and poor to fair (42 percent) when the external skin or orbital contents were excised. Secondary procedures were required in 16 of 50 patients (32.0 percent). Free-tissue transfer provides the most effective and reliable form of immediate reconstruction for complex maxillectomy defects. The rectus abdominis and radial forearm flaps in combination with immediate bone grafting or as osteocutaneous flaps reliably provide the best aesthetic and functional results. An algorithm based on the type of maxillary resection can be followed to determine the best approach to reconstruction. PMID- 10845287 TI - Histomorphologic changes of hair follicles in human expanded scalp. AB - To obtain information about changes that occur in hair follicles when tissue expansion is performed on the scalp, punch biopsy samples were taken from normal scalp (stage I) and the top of the expander immediately before removal (stage II) and 12 weeks after the second operation (expander removal and flap transposition, stage III) in 10 consecutive patients. We compared histologic and quantitative changes of hair follicles in transverse sections of the expanded scalp and long term changes with those in normal controls using three specimens from each patient. Both the proportion of terminal hair to vellus hair and the proportion of anagen hair to telogen hair were significantly increased during stages II and III (p < 0.05). Perifollicular inflammation and fibrosis were observed during stage II but disappeared during stage III. All these findings imply that tissue expansion at the hair-bearing scalp made the telogen period short, possibly because of active epidermal mitosis. PMID- 10845286 TI - Free anterolateral thigh flap for reconstruction of head and neck defects following cancer ablation. AB - Thirty-seven consecutive free anterolateral thigh flaps in 36 patients were transferred for reconstruction of head and neck defects following cancer ablation between January of 1997 and June of 1998. The success rate was 97 percent (36 of 37), with one flap lost due to a twisted perforator. The anatomic variations and length of the vascular pedicle were investigated to obtain better knowledge of anatomy and to avoid several surgical pitfalls when it is used for head and neck reconstruction. The cutaneous perforators were always found and presented as musculocutaneous or septocutaneous perforators in this series of 37 anterolateral thigh flaps. They were classified into four types according to the perforator derivation and the direction in which it traversed the vastus lateralis muscle. In type I, vertical musculocutaneous perforators from the descending branch of the lateral circumflex femoral artery were found in 56.8 percent of cases (21 of 37), and they were 4.83 +/- 2.04 cm in length. In type II, horizontal musculocutaneous perforators from the transverse branch of the lateral circumflex femoral artery were found in 27.0 percent of cases (10 of 37), and they were 6.77 +/- 3.48 cm in length. In type III, vertical septocutaneous perforators from the descending branch of the lateral circumflex femoral artery were found in 10.8 percent of cases (4 of 37), and they were 3.60 +/- 1.47 cm in length. In type IV, horizontal septocutaneous perforators from the transverse branch of the lateral circumflex femoral artery were found in 5.4 percent of cases (2 of 37). They were 7.75 +/- 1.06 cm in length. The average length of vascular pedicle was 12.01 +/- 1.50 cm, and the arterial diameter was around 2.0 to 2.5 mm; two accompanying veins varied from 1.8 to 3.0 mm and were suitable for anastomosis with the neck vessels. Reconstruction of one-layer defect, external skin or intraoral lining, was carried out in 18 cases, through-and-through defect in 17 cases, and composite mandibular defect in two cases. With increasing knowledge of anatomy and refinements of surgical technique, the anterolateral thigh flap can be harvested safely to reconstruct complicated defects of head and neck following cancer ablation with only minimal donor-site morbidity. PMID- 10845288 TI - Physical and psychosocial symptoms among 88 volunteer subjects compared with patients seeking plastic surgery procedures to the breast. AB - In an investigation of the relationship between macromastia and physical and psychosocial symptoms, 88 female university students, 21 augmentation mammaplasty patients, and 31 breast reduction patients graded somatic and psychosocial symptoms. The intent of the study was to discover which complaints were most common among women presenting for reduction mammaplasty and to determine whether height/weight index and brassiere chest measurement and cup size might affect their symptoms. Both the student group and the augmentation mammaplasty patients differed significantly from the breast reduction patients. Eighty-one percent of the reduction patients complained of neck and back pain. Seventy-seven percent complained of shoulder pain, 58 percent complained of chafing or rash; 45 percent reported significant limitation in their activity; and 52 percent were unhappy with their appearance (p < 0.001 compared with augmentation and student groups). Physical symptoms were related to height/weight index and bra chest and cup sizes in each of the three participating groups. It was found that patients who present for symptom-related reduction mammaplasty have a disease-specific group of physical and psychosocial complaints that are more directly related to large breast size than to being overweight. PMID- 10845289 TI - Effect of smoking on complications in patients undergoing free TRAM flap breast reconstruction. AB - Free pedicled transverse rectus abdominis myocutaneous (TRAM) flap breast reconstruction is often advocated as the procedure of choice for autogenous tissue breast reconstruction in high-risk patients, such as smokers. However, whether use of the free TRAM flap is a desirable option for breast reconstruction in smokers is still unclear. All patients undergoing breast reconstruction with free TRAM flaps at our institution between February of 1989 and May of 1998 were reviewed. Patients were classified as smokers, former smokers (patients who had stopped smoking at least 4 weeks before surgery), and nonsmokers. Flap and donor site complications in the three groups were compared. Information on demographic characteristics, body mass index, and comorbid medical conditions was used to perform multivariate statistical analysis. A total of 936 breast reconstructions with free TRAM flaps were performed in 718 patients (80.9 percent immediate; 23.3 percent bilateral). There were 478 nonsmokers, 150 former smokers, and 90 smokers. Flap complications occurred in 222 (23.7 percent) of 936 flaps. Smokers had a higher incidence of mastectomy flap necrosis than nonsmokers (18.9 percent versus 9.0 percent; p = 0.005). Smokers who underwent immediate reconstruction had a significantly higher incidence of mastectomy skin flap necrosis than did smokers who underwent delayed reconstruction (21.7 percent versus 0 percent; p = 0.039). Donor-site complications occurred in 106 (14.8 percent) of 718 patients. Donor-site complications were more common in smokers than in former smokers (25.6 percent versus 10.0 percent; p = 0.001) or nonsmokers (25.6 percent versus 14.2 percent; p = 0.007). Compared with nonsmokers, smokers had significantly higher rates of abdominal flap necrosis (4.4 percent versus 0.8 percent; p = 0.025) and hernia (6.7 percent versus 2.1 percent; p = 0.016). No significant difference in complication rates was noted between former smokers and nonsmokers. Among smokers, patients with a smoking history of greater than 10 pack-years had a significantly higher overall complication rate compared with patients with a smoking history of 10 or fewer pack-years (55.8 percent versus 23.8 percent; p = 0.049). In summary, free TRAM flap breast reconstruction in smokers was not associated with a significant increase in the rates of vessel thrombosis, flap loss, or fat necrosis compared with rates in nonsmokers. However, smokers were at significantly higher risk for mastectomy skin flap necrosis, abdominal flap necrosis, and hernia compared with nonsmokers. Patients with a smoking history of greater than 10 pack-years were at especially high risk for perioperative complications, suggesting that this should be considered a relative contraindication for free TRAM flap breast reconstruction. Smoking-related complications were significantly reduced when the reconstruction was delayed or when the patient stopped smoking at least 4 weeks before surgery. PMID- 10845290 TI - The value of preoperative Doppler sonography for planning free perforator flaps. AB - The individual perforating vessels have a high degree of anatomical variation, therefore it is desirable to conduct a careful examination of them before undertaking a perforator flap operation. Because locating the vessels beforehand makes performing the operative procedure much easier, the aim of the present study was to assess the value of using simple acoustic Doppler sonography to plan a perforator flap operation. The vessel examinations were carried out before taking 46 free microvascular flaps from either the lower abdominal wall or the buttock for reconstructive breast surgery. The perforating vessels located were marked, and their position relative to the umbilicus or the most cranial point of the rima ani recorded using a coordinate system. In 40 patients, a perforator flap operation (deep inferior epigastric perforator flap, n = 32; superior gluteal artery perforator flap, n = 8) was actually carried out; in six of these patients, a myocutaneous flap was used because of the insufficient availability of perforating vessels. Before the operation, perforating vessels were marked for each patient, with an average of 7.3 for the deep inferior epigastric perforator flap and 6.5 for the superior gluteal artery perforator flap. Out of 286 vessels marked for later perforator flaps, 162 were identified during the operation. A preoperatively marked vessel was used in 37 of 40 patients. In the remaining patients, a vessel was used that had not been previously marked. The vertical and horizontal distance between the perforating vessels identified during the operation and the preoperative marks averaged 0.8 cm. The results show preoperative Doppler sonography to be useful for locating the position of individual perforating vessels, making it much easier to find them during the operation. PMID- 10845291 TI - Reconstruction of large sacral defects following total sacrectomy. AB - Total sacrectomies for cancer ablation often result in extensive defects that are challenging to reconstruct. In an effort to elucidate the criteria to select the most effective reconstructive options, we reviewed our experience with the management of large sacral wound defects. All patients who had a sacral defect reconstruction after a total sacrectomy at our institution between January of 1993 and August of 1998 were reviewed. The size of the defect, the type of reconstruction, postoperative complications, and functional outcome in each patient were assessed. A total of 27 flaps were performed in 25 patients for sacral defect reconstruction after a total sacrectomy. Diagnoses consisted of chordoma (n = 13), giant cell carcinoma (n = 2), sarcoma (n = 5), rectal adenocarcinoma (n = 4), and radiation induced necrosis (n = 1). The size of sacral defects ranged from 18 to 450 cm2 (mean, 189.8 cm2). Ten patients, including five who had preoperative radiation therapy, underwent transpelvic vertical rectus abdominis myocutaneous (VRAM) flap reconstruction for sacral defects with a mean size of 203.3 cm2. Of these, five patients (50 percent) had complications (four minor wound dehiscences and one seroma). Eight patients, including one who had preoperative radiation therapy, underwent bilateral gluteal advancement flap reconstruction for sacral defects with a mean size of 198.0 cm2. They had no complications. Two patients, both of whom had preoperative radiation therapy, underwent gluteal rotation flap reconstruction for sacral defects of 120 cm2 and 144 cm2. Both patients had complications (one partial flap loss and one nonhealing wound requiring a free flap). Three patients, including one who had preoperative radiation therapy, underwent reconstruction with combined gluteal and posterior thigh flaps for sacral defects with a mean size of 246 cm2; two of these patients had partial necrosis of the posterior thigh flaps. Three patients, all of whom had preoperative radiation therapy, underwent free flap reconstruction for sacral defects with a mean size of 144.3 cm2. They had no complications. Our experience suggests that there are three reliable options for the reconstruction of large sacral wound defects: bilateral gluteal advancement flaps, transpelvic rectus myocutaneous flaps, and free flaps. In patients with no preoperative radiation therapy and intact gluteal vessels, the use of bilateral gluteal advancement flaps should be considered. In patients with a history of radiation to the sacral area and in patients whose gluteal vessels have been damaged, the use of the transpelvic VRAM flap should be considered. If the transpelvic VRAM flap cannot be used because of previous abdominal surgery, a free flap should be considered as a last option. PMID- 10845292 TI - Flap perfusion after free musculocutaneous tissue transfer: the impact of postoperative complications. AB - In a previous study, the authors found persistence of pedicle blood flow up to 10 years after uncomplicated free latissimus dorsi transfer. In this study, the impact of postoperative complications (hematoma, thrombosis, infection) and successful surgical revision was tested. Since 1982, more than 1200 free tissue transfers have been performed at the authors' institution (Hannover Medical School). Of these, the authors selected two groups of 30 patients each who had received a free latissimus dorsi transfer to the lower leg without microsurgical nerve coaptation for wound coverage. All patients included in this study were carefully selected for clinical homogeneity, with one difference: group I comprised patients who had no postoperative complications after free latissimus dorsi transfer. Group II included only patients with major postoperative complications after the procedure. All flaps in group II survived after successful surgical revision. The arteries, which nourished the lower leg, were visualized and documented by means of a duplex scanner in both groups. Three different time intervals were chosen for measurements of blood flow: 4 to 6 months (groups I.I and II.I), 4 to 6 years (groups I.II and II.II), and 8 to 10 years (groups I.III and II.III). Quantitative measurements of local flap perfusion in milliliters per minute per 100 g tissue were performed by means of the hydrogen clearance technique. In each patient, a total of nine measurements was performed in three phases: phase A, before closing the vascular pedicle by manual compression (n = 3); phase B, with a closed pedicle (n = 3); and phase C, after releasing the vascular pedicle from manual compression (n = 3). Each measurement took approximately 10 minutes. One hundred percent closure of each pedicle in phase B was confirmed by the duplex scanner. Furthermore, all patients were monitored both clinically and by means of the hydrogen clearance technique during phase B for adequate blood supply to the lower leg. Lower leg perfusion showed no statistical differences for phases A, B, and C in all groups of patients. In group I, no statistical differences in local flap perfusion were encountered for phases A and C. In phase B, however, a statistically significant (p < 0.01) complete extinction of local flap perfusion was registered in all patients of group I at the site of the flap's skin paddle. In group II, however, persistent flap perfusion was registered during phase B in up to 50 percent of cases in one subgroup (II.III). No statistically significant alterations of local blood flow were registered in the surrounding tissue of group II during phases A, B, and C. Patients with thrombosis of the venous anastomosis (n = 7) seemed to have the highest incidence of loss of autonomous blood supply through the vascular pedicle (5 out of 11 cases). No inconstant results were found during the repetitive measurements (n = 3) for each patient in each phase. After uncomplicated free tissue transfer, the flap's intact vascular pedicle seems to play an important role in permanent flap survival up to 10 years after the procedure. Postoperative complications after free tissue transfer with successful surgical revision, especially venous thrombosis of the vascular anastomosis, may lead to loss of vascular flap autonomy over time. PMID- 10845293 TI - The lateral forearm flap as a modification of the lateral arm flap: vascular anatomy and clinical implications. AB - The forearm extension of the lateral arm flap was introduced on the basis of the vascular territory of the posterior radial collateral artery extending beyond the elbow into the forearm. However, there is controversy as to whether the posterior radial collateral artery extends as a single trunk below the elbow or if it terminates more proximally with only a rich vascular plexus extending beyond the elbow. The purpose of this study was to revisit the artery's anatomy in the region of the elbow and to study its distribution in the forearm. Using latex and barium-gelatin injections of the posterior radial collateral artery in ten cadaveric upper limbs, it was observed that terminal branching of the artery occurred 4.5 cm proximal to the lateral epicondyle of the humerus. Distal to the epicondyle, the terminal branches of the posterior radial collateral artery were seen to fan out as finely arborized branches supplying the lateral forearm skin. No single, constant vascular trunk to the forearm skin could be identified. Furthermore, in its distribution toward the periphery, the terminal branches of the posterior radial collateral artery took an increasingly superficial course. Proximal to the epicondyle, the vessels lay deep within the subcutaneous fat, whereas distal to the epicondyle, they were very close to skin. These findings suggest that lateral forearm skin cannot be islanded without risk of vascular disruption and that the distally sited flap should include skin proximal to the epicondyle for safety. PMID- 10845294 TI - Sensory recovery and Meissner corpuscle number after toe-to-hand transplantation. AB - This study investigated the possible relationship between sensory recovery and receptor number after toe-to-hand transplantation. Moving two-point discrimination was measured after a course of sensory re-education. Meissner corpuscle number was then quantified by light microscopic examination of multiple sections of glabrous skin obtained at pulp reduction and from site-matched normal toe and fingertip skin. Meissner corpuscle number per millimeter was 0.94 in normal toe skin (n = 4), and 0.37 after toe-to-hand transplantation (n = 34). A significant correlation existed between moving two-point discrimination and Meissner corpuscle number (r = -0.62; p < 0.001). No significant relationship was found between moving two-point discrimination and patient age or interval from injury to reconstruction. The results suggest that despite sensory re-education, there may be a level of receptor reinnervation below which good sensory recovery may not be obtained. PMID- 10845295 TI - Free nonvascularized composite nail grafts: an institutional experience. AB - Free, nonvascularized composite nail grafts have been reported as a successful method to reconstruct nail deformities due to congenital anomalies or traumatic defects. The authors performed a decade review of their experience with nine patients who had had 10 free, nonvascularized composite nail grafts. Patient demographics, mechanism of injury, timing, site of reconstruction, and amount of nail to be replaced were all recorded. Results of nail growth in reconstructed nails were judged as excellent, good, fair, or poor on the basis of the appearance of the nail. The majority of reconstructed nails had half or more of the nail bed replaced. The 10 cases (mean follow-up of 1.8 years) that were reported had two excellent, three good, two fair, and three poor outcomes. There was no apparent relation between the successful outcome of the procedure and patient age, timing of reconstruction, or amount of nail bed replaced. Although the authors' experience suggests the unpredictable nature of this type of graft, it should be considered for patients who desire nail reconstruction and are not candidates for ablative or vascularized nail complex transfer procedures. PMID- 10845296 TI - Growth restriction of cranial sutures in the fetal lamb causes deformational changes, not craniosynostosis. AB - Newborns with in utero cranial vault molding can present with severe forms of plagiocephaly. Intrauterine constraint has been proposed as one cause for craniosynostosis. The purpose of this experiment was to investigate whether rigid plate fixation across a fetal cranial suture, representing a severe form of growth restriction in utero, would lead to cranial suture fusion in a fetal lamb model. Six fetal lambs at 85 to 95 days gestation (term = 145 days) underwent laparotomy, hysterotomy, fetal coronal scalp incision, and miniplate screw fixation across the right coronal suture in utero. Two unoperated twins and four unoperated age-matched lambs were used as controls (n = 12). Animals were killed at both 4 and 8 weeks postoperatively. Fetal head analysis consisted of gross examination, photography, basilar and lateral radiographs, and three-dimensional computed tomographic scans. Cranial suture analysis consisted of imaging by computed tomographic scan (axial and sagittal cuts) and histology of experimentally plated coronal sutures, contralateral nonplated coronal sutures and twin control coronal sutures. Gross examination, radiographs, and three dimensional computed tomographic analysis of heads with cranial suture plating showed ipsilateral forehead flattening, contralateral forehead bossing, superiorly displaced ipsilateral orbital rim, anterolateral projection of ipsilateral malar eminence, and anterior position of the ipsilateral ear point compared with the contralateral side of the same animal and normal controls. There was no change in nasal root, chin point, or predentition occlusal plane. Although analysis of the plated coronal sutures by computed tomographic scans showed diminished width or even stenosis, the histology revealed narrowed but patent experimental coronal sutures at 4 and 8 weeks. Contralateral, nonplated coronal sutures were not only patent, but widened compared with normal control sutures. This finding may have represented compensatory changes in the contralateral coronal suture caused by growth restriction at the plated suture. These data demonstrate that intrauterine growth restriction across a cranial suture caused by compression plate fixation resulted in deformational skull changes, not craniosynostosis. In addition, these data strongly support a role for in utero positional molding secondary to growth restriction in the maternal pelvis as a cause for nonsynostotic plagiocephaly seen in newborns. PMID- 10845297 TI - Neonatal induction of tolerance to skeletal tissue allografts without immunosuppression. AB - Vascularized allogeneic skeletal tissue transplantation without the need for host immunosuppression would increase reconstructive options for treating congenital and acquired defects. Because the immune system of a fetus or neonate is immature, it may be possible to induce tolerance to allogeneic skeletal tissues by alloantigen injection during this permissive period. Within 12 hours after birth, 17 neonatal Lewis rats were injected through the superficial temporal vein with 3.5 to 5 million Brown Norway bone marrow cells in 0.1 ml normal saline. Ten weeks after the injection, peripheral blood from the Lewis rats was analyzed for the presence of Brown Norway cells to determine hemopoietic chimerism. The Lewis rats then received a heterotopic, vascularized limb tissue transplant (consisting of the knee, the distal femur, the proximal tibia, and the surrounding muscle on a femoral vascular pedicle) from Brown Norway rat donors to determine their tolerance to the allogeneic tissue. A positive control group (n = 6) consisted of syngeneic transplants from Lewis rats into naive Lewis rats to demonstrate survival of transplants. A negative control group (n = 6) consisted of Brown Norway transplants into naive Lewis rats not receiving bone marrow or other immunosuppressive treatment. The animals were assessed for transplant viability 30 days after transplantation using histologic and bone fluorochrome analysis. All the syngeneic controls (Lewis to Lewis) remained viable throughout the experiment, whereas all the Brown Norway to Lewis controls had rejected. Ten of the 17 allografts transplanted into bone marrow recipients were viable at 30 days, with profuse bleeding from the ends of the bone graft and the surrounding graft muscle. The percent of chimerism correlated with survival, with 3.31 percent (SD = 1.9) of peripheral blood, Brown Norway chimerism present in the prolonged survival groups and 0.75 percent (SD = 0.5) of Brown Norway chimerism in the rejected graft group. This study demonstrated prolonged survival of allogeneic skeletal tissue without immunosuppression after early neonatal injection of allogeneic bone marrow in a rat model. PMID- 10845298 TI - Arteriovenous malformation of the forehead, anterior scalp, and nasal dorsum. AB - Management of complex and relentless large arteriovenous malformations with long term control and acceptable aesthetic results can be accomplished. This outcome requires selective intra-arterial embolization, judicious surgical resection, composite reconstruction with free tissue transfer, other ancillary procedures, or both, and careful serial follow-up examinations to rule out recurrent or persistent disease. PMID- 10845299 TI - Total soft-tissue reconstruction of the middle and lower face with multiple simultaneous free flaps in a pediatric patient. AB - A 2-year-old boy sustained a massive facial soft-tissue wound secondary to a dog attack. Essentially all the soft tissues of the face were absent, including innervation and intraoral lining. We describe the reconstruction of this defect with five simultaneous free tissue transfers. To our knowledge, this is the first report of five simultaneous free flaps in any patient. PMID- 10845300 TI - Treatment of hostile midline back wounds: an extreme approach. AB - The basic principles of successful wound closure remain the same: careful preoperative evaluation, removal of underlying nonviable tissue, and well vascularized soft-tissue coverage. Many complex or "hostile" back wound closures also require stabilization of the spine and a two-layered wound closure. The use of long arteriovenous fistulas with free tissue transfer provides an additional weapon for the treatment of these complex wounds. PMID- 10845301 TI - Treatment of extensive sternum keloid scar with surgical removal and immediate reconstruction with skin flaps from a combined mammaplasty. AB - We report the case of a 47-year-old woman with a large keloid scar on the sternum who was submitted to a simultaneous scar removal with bilateral breast-reduction mammaplasty. Breast reduction was performed to reduce local skin tension and to provide a skin flap for the full reconstruction of the scar-removal site. The association of these surgical procedures stands as a viable alternative for the reconstruction of the sternum region, producing less keloid scarring. PMID- 10845302 TI - Double-toe transplantation following temporary insertion of a block of silicone for reconstruction of a traumatic metacarpal defect. AB - The treatment of the mutilated hand with more than one digit missing and a metacarpal defect is challenging. We used double-toe transfer after temporary insertion of a block of silicone to fill the bony defect. PMID- 10845303 TI - Endoscopic approach for the resection of forehead masses. AB - Over the past several years, surgery aided by the endoscope has come into favor for a number of reasons. Because it is minimally invasive surgery, it has less morbidity, thus, reduced postoperative pain and complications. It results in earlier mobilization and shorter hospitalization, and most importantly, it contributes to an improved cosmetic appearance as a result of a shortened incision line concealed within the hairline in most cases. We have proposed an alternative approach to the surgical resection of forehead masses by means of the endoscope, which has proven to be useful not only for diagnosis but also as a therapeutic tool for the removal of forehead lesions. This report described the clinical experience with the removal of forehead masses in four patients. The cases illustrated the feasibility and ease of resecting a variety of forehead masses with excellent cosmetic results. We hope that more plastic surgeons will use the proposed technique and will continue to explore the safe limits of endoscopic plastic surgery. PMID- 10845304 TI - V-Y advancement flap in upper-lip reconstruction. PMID- 10845305 TI - The impact of biomolecular medicine and tissue engineering on plastic surgery in the 21st century. PMID- 10845306 TI - Acute burns. AB - Burn injuries are complex cutaneous traumas cared for by many plastic surgeons. Care is stratified by burn size, depth, and associated injuries. Advances in surgical technique, wound care, and bioengineered skin have resulted in excellent outcomes for most burn survivors. Moderate burn injuries can be treated effectively by an interested and experienced plastic surgeon. PMID- 10845307 TI - Strategic planning in plastic surgery: what it can do for you. PMID- 10845308 TI - Nonsurgical breast enlargement using an external soft-tissue expansion system. AB - Less than 1 percent of the women interested in having larger breasts elect to have surgical augmentation mammaplasty with insertion of breast implants. The purpose of this report is to describe and test the efficacy of a nonsurgical method for breast enlargement that is based on the ability of tissues to grow when subjected to controlled distractive mechanical forces. Seventeen healthy women (aged 18 to 40 years) who were motivated to achieve breast enlargement were enrolled in a single-group study. The participants were asked to wear a brassiere like system that applies a 20-mmHg vacuum distraction force to each breast for 10 to 12 hours/day over a 10-week period. Breast size was measured by three separate methods at regular intervals during and after treatment. Breast tissue water density and architecture were visualized before and after treatment by magnetic resonance imaging scans obtained in the same phase of the menstrual cycle. Twelve subjects completed the study; five withdrawals occurred due to protocol noncompliance. Breast size increased in all women over the 10-week treatment course and peaked at week 10 (final treatment); the average increase per woman was 98 +/- 67 percent over starting size. Partial recoil was seen in the first week after terminating treatment, with no significant further size reduction after up to 30 weeks of follow-up. The stable long-term increase in breast size was 55 percent (range, 15 to 115 percent). Magnetic resonance images showed no edema and confirmed the proportionate enlargement of both adipose and fibroglandular tissue components. A statistically significant decrease in body weight occurred during the course of the study, and scores on the self-esteem questionnaire improved significantly. All participants were very pleased with the outcome and reported that the device was comfortable to wear. No adverse events were recorded during the use of the device or after treatment. We conclude that true breast enlargement can be achieved with the daily use of an appropriately designed external expansion system. This nonsurgical and noninvasive alternative for breast enlargement is effective and well tolerated. PMID- 10845309 TI - Human-derived and new synthetic injectable materials for soft-tissue augmentation: current status and role in cosmetic surgery. AB - The recent development of human-derived and new synthetic filling agents heralds a new era in soft-tissue augmentation. Many of the disadvantages of xenogenic and prior exogenous materials have been overcome with the advent of these autologous, allogeneic, and inert synthetic alternatives. Early reports using human-derived and inert exogenous filling agents have demonstrated good results and prolonged correction. It is too early, however, to assess the long-term efficacy of these agents. Future investigations should include histologic examination after facial implantation to document long-term safety and efficacy. PMID- 10845310 TI - A prospective analysis of patients undergoing silicone breast implant explantation. AB - Despite the lack of a scientifically proven link between silicone implants and disease, many women have chosen to have their implants removed out of concern for their health. Unfortunately, there are few studies in the literature that have investigated the outcome of explanations, and there are no prospective analyses of the effect explantation has on a patient's general health. The goal of this study was to use a prospective database to determine whether there were any preoperative parameters that could be used to predict which patients would be improved following removal of silicone breast implants and to provide a quantifiable measure of that improvement. A total of 38 patients with silicone breast implants underwent operative removal of their breast implants by faculty at the University of Texas Southwestern Medical Center. They were given questionnaires regarding several personal and medical parameters to be completed preoperatively, at 6 weeks postoperatively, and at 6 months postoperatively. In addition, their physicians completed preoperative and postoperative evaluations of the patient's general health status. A control group of 38 patients was established; they were matched with the experimental group with regard to age and other initial parameters. Their responses to questionnaires were then grouped according to standard subscales to evaluate physical functioning, physical role, bodily pain, general health, vitality, social functioning, emotional role, mental health, appearance evaluation, appearance orientation, illness orientation, and body area satisfaction. When compared with the control group, we found that patients who had undergone explantation showed a temporary decrease in musculoskeletal symptoms and bodily pain, as well as an increase in vitality, mental health, and body area satisfaction. Of the experimental group, those who initially indicated a higher number of musculoskeletal symptoms and a higher appearance evaluation were more likely to indicate a significant improvement in general health since explantation. PMID- 10845311 TI - Evaluation of cosmetic results of a randomized trial comparing surgery and radiotherapy in the treatment of basal cell carcinoma of the face. AB - Basal cell carcinoma is the most frequent cutaneous carcinoma, and it is characterized by its local spreading and an exceptional tendency to metastasize. Radical excision or destruction ensures the highest chance of cure. The most frequent site of this tumor is the face, where radical excision is limited by the proximity of essential anatomic structures. The main difficulty is to avoid mutilation and to provide good cosmetic results despite the vicinity of the eyes, the nose, and the mouth. Surgery and radiotherapy are known to provide similar chances of cure, but results concerning cosmetic sequelae are controversial, depending sometimes on the specialty of the physician in charge of the treatment. A randomized trial was performed at the Gustave-Roussy Institute to compare basal cell carcinomas of the face treated either by surgery or by radiotherapy. In summary, a significant advantage was observed in favor of surgery, as has been published elsewhere. Looking at the details of the cosmetic results, we analyzed the specific methodology of the cosmetic evaluation set up to eradicate the usual bias owing to subjective judgments. We looked also to the evolution of the cosmetic results with time. A panel of five judges performed repeated evaluations during the follow-up, and standardized photographs were taken at each visit and rated later by three nonmedical judges. In total, 174 patients were treated by surgery and 173 by radiotherapy; the choice of the treatment was allocated by randomization. Postoperative complications were higher in the radiotherapy group. The final cosmetic results after 4 years of follow-up were rated significantly better with surgery than with radiotherapy (good in 87 percent versus 69 percent according to the patient, 79 percent versus 40 percent according to the dermatologist, and respectively for each of the observers). Evolution of the ratings during the follow-up demonstrated an improvement of the cosmesis after surgery and stable or deteriorated results after radiotherapy. The same trend was observed regardless of the site of the tumor on the face, except for the nose, where the difference--still in favor of the surgery--was not significant. Concordance of all assessments in our study was the main guarantee of reliability of our methodology for cosmetic evaluation. PMID- 10845312 TI - Reduction of lower palpebral bulge by plicating attenuated orbital septa: a technical modification in cosmetic blepharoplasty. AB - Bulging of the lower eyelid is regarded as a sign of aging. "Herniation" of the periorbital fat pads is traditionally regarded as the factor responsible for the change. Excision of fat pads, therefore, has been the mainstay of treatment in reducing the palpebral bulge in cosmetic blepharoplasty. The surgical excision of"excess" and "herniated" fat pads, however, causes problems such as lid ecchymosis, chemosis, lid contour irregularity, ectropion, and retrobulbar hematoma formation. The author proposes that the loss of fat pad support caused by the attenuation of the orbital septa, not herniation of the excess fat pads, is the major factor responsible for the bulge. The author further proposes that the functional integrity of the orbital septum can be restored by plicating the attenuated orbital septa with 5-0 absorbable sutures. This technique of invaginating the protruded fat pad was performed in 138 individuals (276 lower eyelids). The operation was technically simple, and the approach was "tissue friendly." The results obtained, with the exception of a mild degree of puffiness encountered soon after the surgery, were satisfactory. Morbidity was minimal. PMID- 10845313 TI - In search of the ideal nose. AB - The proportions of the so-called ideal nasal shape and the operations designed to achieve this have been the subject of innumerable papers. The vast majority of patients in the authors' practices desire a more refined version of the nose and do not want the so-called nosejob look with ideal proportions. A total of 2930 cosmetic rhinoplasties performed over the past 20 years, based on the philosophy of refining the more unrefined features of the nose according to the patients' wishes, have been reviewed, and almost all of these patients were satisfied with the long-term shape of their nose, even though in many cases these noses would be considered less than ideal by some of the newer criteria. PMID- 10845314 TI - Sonography of nasal tip anatomy and surgical tip refinement. AB - The amorphous or wide nasal tip is the most commonly encountered nasal tip deformity, but little has been done to measure the effect of standard rhinoplasty techniques on nasal tip width. In the clinical routine, nasal tip width and soft tissue cover thickness are estimated by inspection and palpation rather than by measurement. In this study, a B-mode sonograph with a 12-MHz transducer was used in a noncontact mode to measure tip width 0.5 cm occipital to the tip defining point, distance between the alar cartilage domes, and thickness of the soft tissue cover overlying the lower lateral cartilages. These parameters were measured 3 to 8 weeks before and 56 days to 19 months after a transdomal suture tip plasty in 18 patients. The distance between the alar cartilage domes seemed to be an important factor for tip width because interdomal distance, not soft tissue cover thickness, correlated with tip width before surgery (correlation: 0.53). Conversely, the degree of tip refinement correlated with preoperative soft tissue cover thickness (correlation: 0.75), but not with interdomal distance. Ultrasonic imaging of nasal soft tissues may help to assess the effect of different tip refining procedures and other soft-tissue changes after rhinoplasty. PMID- 10845315 TI - Progressive tension sutures: a technique to reduce local complications in abdominoplasty. AB - Abdominoplasty has evolved as a very effective and satisfactory procedure, especially when combined with liposuction and the repair of diastasis recti. However, local complications, including hematoma and seroma formation, flap necrosis, and hypertrophic scars, continue to plague this procedure. The authors present a relatively simple and reproducible technique that allows extensive liposuction in conjunction with abdominoplasty; they think this technique reduces the incidence of local complications. This technique, the use of progressive tension sutures, has been used in their practice for more than 15 years. A retrospective review of 65 consecutive abdominoplasty patients demonstrates a very low local complication rate when compared with historical controls. In this series of both full and modified abdominoplasty patients who were followed for an average of 18 months, the authors had no hematomas, seromas, or skin flap necrosis. PMID- 10845316 TI - Traumatic abdominal wall pseudolipoma following suction-assisted lipectomy. PMID- 10845317 TI - Relevance of the lesser occipital nerve in facial rejuvenation surgery. AB - Nerve injuries are possible during facial rejuvenation surgery. The great auricular nerve has been studied; however, little is known about the lesser occipital nerve and its relevance in facial rejuvenation surgery. To understand the importance of the lesser occipital nerve in a face lift procedure, the specific anatomy of the nerve was studied in the laboratory in 19 hemifaces, with additional nerve observations in the operating room. The course of the lesser occipital nerve, its branches, and the relationship with the surrounding structures were evaluated and recorded. The great auricular nerve was also dissected to compare the two nerve territories. In the majority of the dissections, the lesser occipital nerve supplied the superior ear and the mastoid area, whereas the great auricular nerve innervated the inferior ear and a portion of the preauricular area. The nerves, however, were variable in size and distribution. Five lesser occipital nerves provided the dominant supply to the ear, compensating for a small great auricular nerve contribution. Therefore, injury to the lesser occipital nerve can result in a major sensory deficit of the ear. We also found the lesser occipital nerve to have a subcutaneous course at a proximal and variable level. These nerve branches can be superficial, and therefore postauricular flap dissection can injure the nerve if the flap is dissected at the fascial level. We therefore suggest that the dissection be at a more superficial level to avoid nerve injury. And finally, if SMAS/platysma suspension sutures are placed, we suggest these be done in a vertical-oblique direction along the course of the lesser occipital nerve, because this should minimize the possibility of trapping terminal branches. PMID- 10845318 TI - Suction mammaplasty: the use of suction lipectomy to reduce large breasts. PMID- 10845319 TI - The future is yours, my son. PMID- 10845320 TI - Reversed thinking. PMID- 10845321 TI - Periauricular face lift incisions and the auricular anchor. PMID- 10845322 TI - One size fits all: a surgical technique for correction of all types of prominent ears. PMID- 10845323 TI - Sensorial innervation of the depressor anguli oris flap: from the buccal nerve or the mental nerve? PMID- 10845324 TI - Transplantation of fertilized ova of cleft lip and palate-resistant C57BL/6 strain mice to pseudopregnant cleft lip and palate-prone A/J strain mice. PMID- 10845325 TI - The superficial subciliary cheek lift, a technique for rejuvenating the infraorbital region and nasojugal groove: clinical series of 71 patients. PMID- 10845326 TI - Desmoid tumor of the breast after reconstruction with implant. PMID- 10845327 TI - Hemothorax as an unusual complication of reduction mammaplasty. PMID- 10845328 TI - Reason for lack of acceptance of radical breast reduction. PMID- 10845330 TI - Warning about the use of rectus femoris flaps in patients with previous surgery for congenital hip dislocation. PMID- 10845329 TI - Breast reconstruction with latissimus dorsi flap: improved aesthetic results after transection of its humeral insertion. PMID- 10845331 TI - Circle-to-W flap. PMID- 10845332 TI - The expanded suprapubic area as a skin donor site in the treatment of congenital absence of the vagina. PMID- 10845333 TI - Conception following penile reconstruction. PMID- 10845334 TI - Topical lignocaine gel for split-thickness skin graft donor-site pain management. PMID- 10845335 TI - Simple method of designing a bilobed flap. PMID- 10845336 TI - Obsolete PMID- 10845337 TI - Schematic representations. PMID- 10845338 TI - Should aspirin be discontinued two weeks before elective surgery? PMID- 10845339 TI - Digital photography for the plastic surgeon. PMID- 10845340 TI - The role of the mind and emotions of patient and surgeon in the outcome of surgery. PMID- 10845341 TI - Some thoughts on the future of plastic surgery. PMID- 10845342 TI - A psychodynamic view of psychosomatic medicine. AB - OBJECTIVE: The objective of this article is to assess the relevance of psychodynamic observations and theory for psychosomatic medicine. METHODS: The evolution of the psychodynamic formulation of psychosomatic symptom formation is described in a brief historical review. RESULTS: There are two distinctly different pathways along which stress-induced psychological arousal is transformed into somatic symptoms. CONCLUSIONS: Psychodynamic observations and theory have important implications for psychosomatic research and treatment. PMID- 10845344 TI - Effects of coping on health outcome among women with gastrointestinal disorders. AB - BACKGROUND: Studies have shown that the nature and quality of coping may positively or negatively affect health outcome; however, this relationship has not been well studied among patients with gastrointestinal (GI) disorders. OBJECTIVES: The primary objective was to study the effect of different coping strategies on the health outcome of women with GI disorders and how these coping strategies may modify the effects of education, GI disease type, neuroticism, and abuse severity on health outcome. METHODS: We followed 174 patients in a referral GI clinic for 12 months to assess their health status as a derived variable of daily pain, bed disability days, psychological distress, daily dysfunction, number of visits to physicians, and number of surgeries and procedures. We obtained at baseline their GI diagnosis (functional vs. organic), neuroticism score (NEO Personality Inventory), sexual and/or physical abuse history, and scores on two coping questionnaires. Regressions analyses were used to determine the relative effect of the coping measures on health outcome and their modifying effects on education, GI disease type, neuroticism, and abuse severity. RESULTS: A higher score on the Catastrophizing scale and a lower score on the Self Perceived Ability to Decrease Symptoms scale (Coping Strategies Questionnaire) predicted poor health outcome. Less education, a functional GI diagnosis, a higher neuroticism score, and greater abuse severity also contributed to poor health status. However, the effect of GI disease type and neuroticism on health outcome was significantly reduced by the coping measures. CONCLUSIONS: Maladaptive coping (eg, catastrophizing) and decreased self-perceived ability to decrease symptoms may adversely affect health outcome and may modify the effect of GI disease type and neuroticism on health outcome. PMID- 10845343 TI - Cognitive-behavioral stress management reduces serum cortisol by enhancing benefit finding among women being treated for early stage breast cancer. AB - OBJECTIVE: This study examined the effects of a cognitive-behavioral stress management (CBSM) group intervention on serum cortisol levels in women being treated for stage I or II breast cancer. METHODS: Participants were randomly assigned to undergo a 10-week intervention (N = 24) within 8 weeks after surgery or were placed on a waiting list (N = 10). Cortisol was assessed by means of a radioimmunoassay of blood samples collected at the same time of day just before the start of the intervention and immediately after its completion. The women also reported the degree to which breast cancer had made positive contributions to their lives. RESULTS: Intervention participants showed increased benefit finding and reduced serum cortisol levels, whereas control subjects experienced neither change. Path analysis suggested that the effect of CBSM on cortisol was mediated by increases in benefit finding. CONCLUSIONS: These findings suggest that positive growth enhanced during a time-limited intervention can influence physiological parameters such as cortisol among women with early stage breast cancer. PMID- 10845345 TI - Illness fears in the general population. AB - OBJECTIVES: Because relatively little is known about illness fears, we sought to estimate the prevalence, risk factors, and morbidity associated with such fears in the community. METHODS: We conducted a brief telephone survey of persons aged 40 to 65 years from randomly selected households in the Johnson County, Iowa, area. Respondents were asked whether a series of illness and medical care items made them no more nervous, somewhat more nervous, or much more nervous than other people. Those who reported more discomfort were asked to what extent this interfered with medical care or caused impairment or distress. Information about demographic and health characteristics was also obtained. RESULTS: Five hundred persons, 62% of those contacted, responded to the survey. A factor analysis revealed four fear dimensions: illness/injury, medical care, blood/needle, and aging/death. Five percent of respondents reported much more nervousness in relation to at least four of six illness/injury items, 4% indicated that such fears interfered with their medical care, and 5% reported some negative effect on their life. Similarly, 5% of respondents reported much more nervousness in relation to at least two of four medical care items. Illness/injury fears were somewhat more common in persons with lower income and education and in those with medical conditions. CONCLUSIONS: This survey shows that fears of illness and medical care are common in the general population and indicates that lower socioeconomic status and experience with illness are associated with these fears. The findings also suggest that interference with care occurs among those with the strongest fears. PMID- 10845346 TI - Platelets and psychiatry: lessons learned from old and new studies. AB - Platelets play an important role not only in hemostasis but also in the pathophysiology of coronary artery disease. The complex interactions among the vascular endothelium, platelets, and blood components are one of the most exciting research areas today. This review addresses some fundamentals of platelet physiology and examines why platelets are interesting probes for neurophysiology. Results of current studies suggest that platelets are affected by diverse stressors, including psychological ones, and that platelets offer an interesting vantage point for understanding the neurophysiology of various psychiatric disorders. We also describe how platelets have been used for various types of research, including studies of stress associated with cardiovascular disease and studies of platelets in psychopharmacological research. Finally, we examine some of the psychiatric literature related to platelets; these studies range from case studies from the 1920s to contemporary experimental studies. PMID- 10845347 TI - Social support and salivary cortisol in women with metastatic breast cancer. AB - OBJECTIVE: This study used a cross-sectional design to examine the relationships between social support, both quantity (number of people) and quality (appraisal, belonging, tangible, and self-esteem), and neuroendocrine function (mean and slope of diurnal salivary cortisol) among women with metastatic breast cancer. METHODS: Participants (N = 103) were drawn from a study (N = 125) of the effects of group therapy on emotional adjustment and health in women with metastatic breast cancer. They completed the Interpersonal Support Evaluation List and the Yale Social Support Index and provided saliva samples for assessment of diurnal cortisol levels on each of 3 consecutive days. Diurnal mean levels were calculated using log-transformed cortisol concentrations, and the slope of diurnal cortisol variation was calculated by regression of log-transformed cortisol concentrations on sample collection time. RESULTS: Mean salivary cortisol was negatively related to the Interpersonal Support Evaluation List subscales of appraisal, belonging, and tangible social support. No association was found between quantitative support or the esteem subscale of the Interpersonal Support Evaluation List and mean salivary cortisol. Measures of qualitative and quantitative social support were not associated with the diurnal cortisol slope. CONCLUSIONS: Results show that greater quality of social support is associated with lower cortisol concentrations in women with metastatic breast cancer, which is indicative of healthier neuroendocrine functioning. These results may have clinical implications in the treatment of breast cancer. PMID- 10845348 TI - Do the daily experiences of healthy men and women vary according to occupational prestige and work strain? AB - OBJECTIVE: This study evaluated the influence of occupational prestige and work strain on mood, the occurrence of interpersonal conflict, and ambulatory blood pressure and heart rate. METHODS: Participants were 50 men and 50 women matched for occupational prestige who were healthy and middle-aged and who completed measures of mood and conflict simultaneously with measures of ambulatory blood pressure and heart rate recorded every 30 minutes during waking hours of two workdays and one nonworkday; at the end of each day, overall ratings were made. Work strain was assessed by the Work Section of the Self-Evaluation and Social Support Interview Schedule. Multiple level random regression coefficients analyses were conducted. RESULTS: Men and women with low-prestige occupations experienced more interpersonal conflict, b = -0.03, p = .04, and higher ambulatory heart rate, b = -4.83, p = .004, throughout the three days of the study. Relative to those with low work strain, those reporting high work strain experienced negative emotion, b = -0.41, p < .0001, and boredom, b = -0.17, p < .0004. End of the day ratings of negative mood were more influenced by work strain among men than among women. No effects of occupational prestige or work strain were obtained for ambulatory blood pressure readings after adjustment for physical activity, posture, and location. CONCLUSIONS: Individuals in low prestige occupations experience greater exposure to interpersonal conflict and arousal as indexed by heart rate, which might increase risk for stress-related illnesses often associated with social class. Individuals who report work strain experience negative mood and boredom, both at work and at home. The absence of work effects on ambulatory blood pressure may be due to the participants being healthy. PMID- 10845349 TI - Gender differences in processing information for making self-assessments of health. AB - OBJECTIVE: This study proposes that women's greater inclusiveness of various sources of information when making self-assessed health (SAH) judgments accounts for the finding that SAH is a weaker predictor of mortality in women than in men. METHODS: Data from a sample of 830 elderly residents of a retirement community and a 5-year mortality follow-up study were used to examine the bases for women's and men's reports of negative affect (NA) and judgments of SAH. The degree to which each health-related measure accounts for the SAH-mortality association in each gender group was examined. RESULTS: The findings support two possible explanations for the lower accuracy of SAH as a predictor of mortality among women: 1) In both men and women, NA is associated with poorer SAH, but in men, NA is more closely linked to serious disease in conjunction with other negative life events, whereas in women, NA reflects a wider range of factors not specific to serious disease. 2) Men's SAH judgments reflect mainly serious, life-threatening disease (eg, heart disease), whereas women's SAH judgments reflect both life threatening and non-life-threatening disease (eg, joint diseases). CONCLUSIONS: Women's SAH judgments and NAs are based on a wider range of health-related and non-health-related factors than are men's. This difference can explain gender differences in the accuracy of SAH judgments and may be related to other documented differences in women's physical and mental health and illness behavior. The findings emphasize the need to study the bases of NA and other self evaluations separately for women and men. PMID- 10845351 TI - The role of stressors and psychosocial variables in the stress process: a study of chronic caregiver stress. AB - OBJECTIVE: An investigation was conducted 1) to examine the relative importance of stressor types (ie, daily hassles, caregiving-specific stressors, and life events) on the stress response, 2) to assess the stability of relationships between psychosocial variables and stress over a 6-month period, and 3) to explore how the nature and magnitude of the contributions made by stressors and psychosocial factors to the stress process varied according to the qualitative characteristics of the stress response (ie, anxiety, depression, and stress). METHODS: Fifty spousal caregivers of patients with dementia were recruited and asked to participate in a detailed psychosocial evaluation at 3-month intervals; the evaluation involved measurement of stressor frequency, psychosocial variables, and indices of the stress response (ie, anxiety, depression, and stress). RESULTS: The data revealed that the effects of stressors and psychosocial factors on the stress response were considerable (accounting for 49 63% of the variance in stress response measures). Furthermore, there was some evidence of stability in the effects of the stressor and mediator variables on the stress response. Specifically, the contributions of life events and caregiver difficulties were largely consistent at both 3 and 6 months, and the psychosocial factor of "reactive coping and self-appraisal" influenced all three stress response indices at both 3 and 6 months. CONCLUSIONS: There is some evidence of stability in the effects of stressors and psychosocial variables on the stress process over a 6-month period. However, it would also seem that the nature of the stress process differs according to the qualitative characteristics of the stress response. PMID- 10845350 TI - Videotape preparation of patients before hip replacement surgery reduces stress. AB - OBJECTIVE: Elective surgery represents a considerable source of stress for the patient. Many attempts have been made to prepare patients before surgery with the aim of reducing stress and improving outcome. This study used a novel approach to fulfill this aim by showing a videotape of a patient undergoing total hip replacement surgery, covering the time period from hospital admission to discharge, that strictly keeps to the patient's perspective. METHODS: Before elective total hip replacement surgery, 100 patients were randomly assigned to a control group or a preparation group; the latter group was shown the videotape on the evening before surgery. Anxiety and pain were evaluated daily for 5 days, beginning with the preoperative day, by means of the State-Trait Anxiety Inventory and a visual analog scale. Intraoperative heart rate and blood pressure, as well as postoperative intake of analgesics and sedatives, were recorded. Urinary levels of cortisol, epinephrine, and norepinephrine were determined in 12-hour samples collected at night for 5 nights, beginning with the preoperative night. RESULTS: Compared with the control group, the preparation group showed significantly less anxiety on the morning before surgery and the mornings of the first 2 postoperative days, and significantly fewer of them had an intraoperative systolic blood pressure increase of more than 15%. The pain ratings did not differ significantly between the two groups, but the prepared patients needed less analgesic medication after surgery. Prepared patients had significantly lower cortisol excretion during the preoperative night and the first 2 postoperative nights. Excretion of catecholamines did not differ significantly between groups. CONCLUSIONS: We conclude that use of the videotape decreased anxiety and stress, measured in terms of urinary cortisol excretion and intraoperative systolic blood pressure increase, in patients undergoing hip replacement surgery and prepared them to cope better with postoperative pain. PMID- 10845352 TI - Active music therapy in Parkinson's disease: an integrative method for motor and emotional rehabilitation. AB - BACKGROUND: Modern management of Parkinson's disease (PD) aims to obtain symptom control, to reduce clinical disability, and to improve quality of life. Music acts as a specific stimulus to obtain motor and emotional responses by combining movement and stimulation of different sensory pathways. We explored the efficacy of active music therapy (MT) on motor and emotional functions in patients with PD. METHODS: This prospective, randomized, controlled, single-blinded study lasted 3 months. It consisted of weekly sessions of MT and physical therapy (PT). Thirty-two patients with PD, all stable responders to levodopa and in Hoehn and Yahr stage 2 or 3, were randomly assigned to two groups of 16 patients each. We assessed severity of PD with the Unified Parkinson's Disease Rating Scale, emotional functions with the Happiness Measure, and quality of life using the Parkinson's Disease Quality of Life Questionnaire. MT sessions consisted of choral singing, voice exercise, rhythmic and free body movements, and active music involving collective invention. PT sessions included a series of passive stretching exercises, specific motor tasks, and strategies to improve balance and gait. RESULTS: MT had a significant overall effect on bradykinesia as measured by the Unified Parkinson's Disease Rating Scale (p < .034). Post-MT session findings were consistent with motor improvement, especially in bradykinesia items (p < .0001). Over time, changes on the Happiness Measure confirmed a beneficial effect of MT on emotional functions (p < .0001). Improvements in activities of daily living and in quality of life were also documented in the MT group (p < .0001). PT improved rigidity (p < .0001). CONCLUSIONS: MT is effective on motor, affective, and behavioral functions. We propose active MT as a new method for inclusion in PD rehabilitation programs. PMID- 10845353 TI - Physiological stress responses in Filipino-American immigrant nurses: the effects of residence time, life-style, and job strain. AB - OBJECTIVE: The purpose of this study was to evaluate the relationship between measures of Americanization (the adoption of American life-style and culture) and physiological measures of stress in Filipino-American immigrants. METHODS: Ambulatory blood pressure monitors and timed urine collections were used to evaluate blood pressure and urinary catecholamine excretion across the work, home, and sleep daily settings among 31 healthy, premenopausal, immigrant Filipino-American women employed as nurses or nurse's aides. Migration history and life-style were evaluated from questionnaire responses. Reported job strain, decision latitude, and psychological demand were obtained from the Job Content Questionnaire. RESULTS: Immigrants who had lived longer in the United States had elevated norepinephrine levels in the work and home settings (p < .05), higher diastolic blood pressure during sleep (p < .01), and lower dips in blood pressure during sleep (p < .05). Job strain measures were not related to blood pressure, catecholamine excretion rates, or residence time in the United States. CONCLUSIONS: The results suggest that indicators of stress increase as a function of time since immigration, although this result is not explained by self-reports of identification with Filipino or American life-style or by measures of job strain. PMID- 10845354 TI - Emotions and stress increase respiratory resistance in asthma. AB - OBJECTIVES: Clinical reports suggest that various emotions and types of stress can precipitate asthmatic symptoms, but there is little experimental evidence to substantiate this claim. We studied the impact of different emotional states and stress on respiratory resistance in asthmatic and nonasthmatic individuals. METHODS: Participants (24 asthmatic and 24 nonasthmatic patients) viewed short film sequences selected to induce anxiety, anger, depression, elation, happiness, contentment, or a neutral affective state and completed two stressful tasks, mental arithmetic to induce active coping efforts and viewing of medical slides to induce passive coping efforts. Oscillatory resistance, heart rate, blood pressure, baroreflex sensitivity, skin conductance level, respiration rate and volume, and self-reported affective state were measured throughout the session. RESULTS: Uniform increases in oscillatory resistance were found in all emotional states compared with the neutral state and during mental arithmetic in both groups. Asthmatic patients showed stronger reactions to the medical slides than healthy control subjects, with significant increases in oscillatory resistance, blood pressure, skin conductance level, and minute volume, as well as higher levels of self-reported depression, arousal, and shortness of breath. Changes in oscillatory resistance were inconsistently correlated with other physiological indices. CONCLUSIONS: Various emotional states and stress increase oscillatory resistance largely independently of concurrent increases in autonomic or ventilatory activity. The particular sensitivity of asthmatics to passive coping demand requires additional research. PMID- 10845355 TI - Psychological effects of HAART: a 2-year study. AB - OBJECTIVE: The objectives of this study were to evaluate the psychological consequences of combination antiretroviral treatment in terms of mood, hope, and life satisfaction in men with symptomatic human immunodeficiency virus (HIV) infection or acquired immune deficiency syndrome and to compare those whose health improved with those whose health did not improve. METHODS: One hundred seventy-three HIV+ gay or bisexual men with symptomatic HIV illness (40% nonwhite) were evaluated semiannually in a university-affiliated research program between July 1995 and December 1997. The primary outcome measures were the Structured Clinical Interview for DSM-IV, Beck Depression Inventory, Endicott Quality of Life Enjoyment and Satisfaction Questionnaire, and Beck Hopelessness Scale. RESULTS: Psychological distress in this sample was mild to moderate at baseline. During the first 2 years that highly active antiretroviral therapy became widely available, we observed a statistically significant but clinically modest reduction in distress in the sample as a whole, with significant covariates of CD4 cell count, HIV symptoms, and social support in a mixed-effects model. Rates of clinical depression declined. However, this generalized mental health improvement was not related to individual medical improvement of markers of HIV illness progression; those classified as improved were no more likely than those who remained unimproved to report greater declines in measures of distress and hopelessness. Number of self-reported physical symptoms were directly related to distress levels. CONCLUSIONS: A cohort effect was observed, with overall psychological improvement. Physical symptoms were more strongly related to psychological distress than were laboratory markers. Consequently, those whose CD4 cell count and HIV RNA viral load reflected successful treatment were no more likely than others to be relieved of the psychological burdens of illness. PMID- 10845356 TI - Urinary catecholamines and cortisol in recent-onset posttraumatic stress disorder after motor vehicle accidents. AB - OBJECTIVE: This study examined relationships among stress hormone levels, posttraumatic stress disorder (PTSD) diagnosis and symptoms, and gender shortly after a common civilian trauma. METHODS: Levels of catecholamines and cortisol in 15-hour urine samples were examined in 55 adults who had been in serious motor vehicle accidents and in 22 age-matched control. RESULTS: Catecholamines were related to PTSD diagnosis and symptoms, but only among men: PTSD-symptomatic men who had been in an accident exhibited elevated levels of epinephrine and norepinephrine 1 month after the accident and had higher epinephrine levels 5 months later. Intrusive thoughts predicted catecholamine levels at 1 month, and avoidance of trauma-relevant stimuli was associated with higher epinephrine levels 5 months later. These effects were not significant among women. Urinary cortisol was also elevated among PTSD-symptomatic men, but not women, and only immediately (1 month) after the accident. For men and women, greater emotional numbing predicted a lower cortisol level 6 months after the accident. CONCLUSIONS: These findings were interpreted as limited support for the generalizability of findings in men with chronic, combat-related PTSD and indicate the need for additional research on psychoendocrine assessment of traumatized women and specific dimensions of PTSD symptomatology. PMID- 10845357 TI - Patient-by-treatment context interaction in chronic disease: a conceptual framework for the study of patient adherence. AB - OBJECTIVE: Previous reviews have concluded that there is little or no evidence supporting a predictable association between patient characteristics and regimen adherence in chronic illness. The primary objective of this article is to propose an alternative conceptual framework for the interpretation and design of adherence research. METHODS: The author's research involving adherence among patients with end-stage renal disease is reviewed and used to illustrate the patient-by-context interactive framework. RESULTS: Adherence is most favorable when the patient's characteristic or preferred style of coping with illness and treatment-related experiences is congruent with the contextual features or demands of the particular type of medical intervention the patient is undergoing. Among patients with end-stage renal disease, a more vigilant or active style of coping is associated with more favorable adherence only for patients undergoing home-based dialysis treatment modalities that are highly patient directed. Among patients receiving hospital-based, provider-controlled treatment, a less vigilant or more passive coping style is associated with more favorable dialysis regimen adherence. CONCLUSIONS: The patient-by-context interactive perspective can provide a useful framework for the interpretation and design of adherence research. PMID- 10845358 TI - Significant other responses are associated with fatigue and functional status among patients with chronic fatigue syndrome. AB - OBJECTIVE: The predictive power of partners' responses to illness behavior for illness outcomes was investigated among couples in which one person had chronic fatigue syndrome (CFS). METHODS: One hundred nineteen participants who met case definition criteria for CFS and were living with a significant other (SO) completed self-report measures of relationship satisfaction, responses of their SO to fatigue symptoms, and outcome measures of fatigue and functional status. RESULTS: The results indicated that more frequent solicitous SO responses to illness behavior were predictive of greater fatigue-related severity and bodily pain. Solicitous SO responses to fatigue behavior were particularly influential in the context of a satisfactory relationship. In highly satisfactory relationships, solicitous SO responses were associated with significantly greater fatigue severity and fatigue-related disability than in relationships characterized by low or average satisfaction. CONCLUSIONS: Solicitous SO responses to CFS-related symptoms are associated with poorer patient outcomes, especially in the context of a satisfactory intimate relationship. Because of the cross-sectional nature of the study, the direction of effects cannot be interpreted unambiguously. SOs may be inadvertently positively reinforcing illness-related behavior: Solicitous partners may help the patient more with tasks of daily living, thereby decreasing the patient's activity level, which may lead to deconditioning and disability. Alternatively, patients with more severe symptoms and disability may present more opportunities for concerned SO responses, which again may be heightened in the context of a caring, satisfactory relationship. In either case, the results suggest that additional research on the role of solicitous SO responses is warranted. PMID- 10845359 TI - Stress and coping and behavioral organization. AB - This article presents a new integrative theoretical framework for stress and coping centered on the concept of behavioral organization and the notions that stress disorganizes behavior and coping aims at reorganizing it. Functional analysis of behavioral organization leads to identification of four core domains of control and to a functional classification of types of stressful conditions, types of negative emotions, and types of coping functions linked directly to the control domains. The control domains are represented cognitively by four postulated kinds of beliefs or views people hold with respect to undertaking an action: performance, process, prospect, and profit beliefs. The four basic control beliefs form the core of the present coping belief model of coping. Recognition of the underlying structure of behavior, and of functional categories of stress and coping related to it, can advance theory and research on stress and coping by enriching its psychological content, improving research designs, and leading to reexamination, reinterpretation, and integration of findings existing in the literature that presently are fragmented and unconnected. PMID- 10845360 TI - Sertraline in diabetic neuropathy: response and biology. PMID- 10845361 TI - Adhesion forces measured by atomic force microscopy in humid air AB - Adhesive forces measured with an atomic force microscope under ambient conditions are generally regarded to be dominated by non-surface-specific capillary force. In this study, the nature of the "pull-off" force on a variety of surfaces was investigated as a function of relative humidity. The results indicate that even under the condition where capillary condensation occurs there is chemical specificity in the measured pull-off force. Issues such as tip-surface contact time and surface roughness were ruled out as possible artifacts. A mathematical model of pull-off force as a function of relative humidity is proposed in which the chemical specificity is explained. PMID- 10845362 TI - Imparting biomimetic ion-gating recognition properties to electrodes with a hydrogen-bonding structured core-shell nanoparticle network. AB - This paper presents findings of the creation of biomimetic ion-gating properties with core-shell nanoparticle network architectures. The architectures were formed by hydrogen-bonding linkages via an exchange-cross-linking-precipitation reaction pathway using gold nanoparticles capped with thiolate shell and alkylthiols terminated with carboxylic groups as model building blocks. Such network assemblies have open frameworks in which void space is in the form of a channel or chamber with the nanometer-sized cores defining its size, the geometric arrangement defining its shape, and the shell structures defining its chemical specificity. The formation of the network linkages via head-to-head hydrogen bonded carboxylic terminals and the reversible pH-tuned structural properties between neutral and ionic states were characterized using infrared reflectance spectroscopic technique. The biomimetic ion-gating properties were demonstrated by measuring the pH-tuned network "open-close" responses to charged redox probes. Such redox responses were shown to depend on the degree of protonation deprotonation of carboxylic groups at the interparticle linkages, core sizes of the nanoparticles, and charges of the redox probes. Differences in structural networking, pH-tuning, and electrochemical gating properties were identified between the network films derived from nanoparticles of two different core sizes (2 and 5 nm). The mechanistic correlation of these structural properties was discussed. These findings have added a new pathway to the current approaches to biomimetic molecular recognition via design of core-shell nanoparticle architectures at both nanocrystal and molecular scales. PMID- 10845363 TI - Design and synthesis of a more highly selective ammonium ionophore than nonactin and its application as an ion-sensing component for an ion-selective electrode AB - A novel ammonium ionophore, which exhibits superior NH4+ selectivity compared with that of the natural antibiotic nonactin, was successfully designed and synthesized based on a 19-membered crown compound (TD19C6) having three decalino subunits in the macrocyclic system. This bulky decalino subunit is effective for (1) increasing the structural rigidity of the cyclic compound, (2) introducing the "block-wall effect", which prevents forming a complex with a large ion, and (3) increasing the lipophilicity of the ionophore molecule. In the ammonium ionophore design, the first factor contributes to increasing the NH4+ selectivity relative to smaller ions such as Li+, Na+, or even the closest size, K+, and the second factor increases the NH4+ selectivity over larger ions such as Rb+ and Cs+. The X-ray structural analysis proved that TD19C6 forms a size-fit complexwith NH4+ in its crown ring cavity. As an application of this ionophore, an ion sensor (ion-selective electrode) was prepared, which exhibited NH4+ to K+ and Na+ selectivity of 10 and 3,000 times, respectively. This electrode showed a better performance compared to the electrode based on nonactin, which is the only ammonium ionophore presently used in practical applications. PMID- 10845364 TI - Amperometric sensing of ethylene oxide in the gas phase. AB - The sensing device used is based on a porous Pt electrode, which is supported on an ion-exchange membrane and directly exposed to the gas phase. Under acid conditions, ethylene oxide was found to be oxidized on the platinum oxide surface at +550 mV vs MSE, thus enabling its monitoring via the measurement of the associated current. A detection limit of 15 ppb was obtained, based on a signal to-noise ratio of three, and a linear dynamic range was found up to 100 ppm. The effects of mass transport, humidity, and oxygen on the cell response, as well as the cross sensitivity to other organic vapors and inorganic gases are discussed. PMID- 10845365 TI - Amperometric detection of histamine with a methylamine dehydrogenase polypyrrole based sensor. AB - Methylamine dehydrogenase (MADH) has been immobilized in a polpyrrole (PPy) film on an electrode surface and used as an amine sensor for the determination of primary amines. Its response to histamine has been characterized in detail. The PPy film containing MADH was formed electrochemically on a gold minielectrode (1 mm diameter) in the presence of ferricyanide. The film was then coated with Nafion. This enzyme electrode did not require any additional cofactors and was not sensitive to oxygen. It exhibited a maximum response current to histamine at applied potentials of 0.24-0.33 V and at pH 7.5-8.5. This MADH-PPy sensor exhibited a response time of less than 3 s. The immobilized MADH on the electrode exhibited Michaelis-Menten behavior similar to that of the free enzyme in solution with a Km value of 1.3 mM. This sensor could be used to reliably detect histamine over a concentration range from approximately 25 microM to 4 mM. This is the first example of a biosensor that uses an immobilized enzyme that possesses the tryptophan tryptophylquinone prosthetic group. PMID- 10845366 TI - Detection of short-lived electrogenerated species by Raman microspectrometry AB - Raman microprobe spectrometry has been applied to the characterization of unstable species generated electrochemically at a microelectrode (radius in the 10 microm range). The ability of the spectroelectrochemical method to detect short-lived intermediates is directly related to its capability to probe small volumes. Raman microprobe spectrometry is appropriate for electrochemical applications because it allows the analysis of approximately 1 microm3 of solution. In spectroelectrochemical experiments, such a volume corresponds to a reaction layer of 1 microm thickness. Potentially, this technique can allow the observation of species with lifetimes of the order of 1 ms. To enhance the capabilities of this spectroscopic technique, we utilized it in combination with steady-state voltammetry at a microelectrode, to increase the concentration of unstable intermediates near the electrode surface. To determine the detection limit of this combined technique, we varied the base concentration as a means for varying the lifetime the radical cation electrogenerated from 9,10 dichloroanthracene. Well-resolved resonance Raman spectra were obtained for this radical cation when the lifetime was > or = 0.1 ms. This short time resolution achieved with micro-Raman spectroelectrochemistry makes this technique a powerful tool for the characterization of short-lived intermediates that are generated electrochemically in solution. PMID- 10845367 TI - On-site analysis of arsenic in groundwater using a microfabricated gold ultramicroelectrode array AB - Rapid on-site analysis of arsenic in groundwater was achieved with a small battery-powered unit in conjunction with a microfabricated gold ultramicroelectrode array (Au-UMEA). The sensor, consisting of 564 UME disks with a unique gold surface created by electron beam evaporation, was demonstrated to be highly sensitive to low-ppb As3+ using square wave anodic stripping voltammetry. The influence of the square wave frequency, pulse amplitude, and deposition potential on the arsenic peak stripping current was investigated. Varying those theoretical parameters yielded results surprisingly similar to those for the thin Hg film case. The performance of the Au-UMEA was evaluated for reproducibility and reliability. Three stability tests showed an average relative standard deviation of 2.5% for 15 consecutive runs. Limits of detection were investigated, and 0.05 ppb As3+ could be measured while maintaining a S/N of 3:1. Interference studies were performed in the presence of 50-500 ppb of Cu2+, Hg2+, and Pb2+. On-site analysis of groundwater containing arsenic was performed with a small battery-powered potentiostat. Quantification was done through standard additions, and these results were compared to the standard EPA methodology. PMID- 10845368 TI - Raman spectroscopic imaging markers for fatigue-related microdamage in bovine bone. AB - Raman spectroscopic markers have been determined for fatigue-related microdamage in bovine bone. Microdamage was induced using a cyclic fatigue loading regime. After loading, the specimens were stained en-bloc with basic fuchsin to facilitate damage visualization and differentiate fatigue-induced damage from cracks generated during subsequent histological sectioning. Bone tissue specimens were examined by light microscopy and hyperspectral near-infrared Raman imaging microscopy. Three regions were defined-tissue with no visible damage, tissue with microcracks, and tissue with diffuse damage. Raman transects, lines of 150-200 Raman spectra, were used for initial tissue surveys. Exploratory factor analysis of the transect Raman spectra has identified spectroscopically distinct chemical microstructures of the bone specimens that correlate with damage. In selected regions of damage, full hyperspectral Raman images were obtained with 1.4-microm spatial resolution. In regions of undamaged tissue, the phosphate nu1 band is found at 957 cm(-1), as expected for the carbonated hydroxyapatic bone mineral. However, in regions of visible microdamage, an additional phosphate nu1 band is observed at 963 cm(-1) and interpreted as a more stoichiometric, less carbonated mineral species. Raman imaging confirms the qualitative relationship between the Raman spectral signature of bone mineral and the type of microdamage in bovine bone. Two tentative explanations for the presence of less carbonated phosphate in damaged regions are proposed. PMID- 10845369 TI - Development of an immunochemical technique for the analysis of trichlorophenols using theoretical models. AB - An immunoassay has been developed for trichlorophenol analysis on the basis of theoretical chemistry modeling studies. These data have allowed us to choose the optimum chemical structure of the immunizing hapten according to realistic similarities with the target analyte. The synthesis of this hapten and the subsequent application of an appropriate immunization protocol have lead to the production of polyclonal antibodies against the target analyte. A homologous direct competitive ELISA has been developed that can be carried out in about 1 h. It has a limit of detection of 0.2 +/- 0.06 microg/L (1.01 +/- 0.3 nM) and it has been proven to tolerate a wide range of ionic strengths and pH values. Thus, the assay has acceptable features in samples with ionic strength between 4 and 56 mS/cm and pH values between 5.5 and 9.5. Studies on the selectivity of this immunoassay have demonstrated a high recognition of the corresponding brominated analogues. Other phenolic compounds do not interfere significantly in the analysis of 2,4,6-trichorophenol using this immunochemical technique. The accuracy of the assay has been evaluated using certified and spiked samples. PMID- 10845370 TI - Design and implementation of a new electrodynamic ion funnel. AB - A new electrodynamic (rf) ion funnel has been developed and evaluated for use in the interface regions (at approximately 1-10 Torr) of atmospheric pressure ion sources (e.g., electrospray ionization (ESI) for mass spectrometry). The ion funnel consists of a ring electrode ion guide with decreasing i.d. and with a superimposed dc potential gradient along the ring stack. The thicknesses of the ring electrodes and the spacings between them were reduced to 0.5 mm from 1.59 mm compared to those used for previous designs. The new ion funnel displays a significant improvement in low-mass transmission (m/z >200) and sensitivity compared to previous designs. The transmission efficiencies for electrosprayed peptides and proteins (ranging in mass from 200 to 17,000 Da) were typically 50 60% of total incoming currents from a heated capillary inlet. The transmitted ion currents were a factor of 30-56 greater than those of the standard interface for peptide samples and a factor of 18-22 greater than those for protein samples. The sensitivity gains realized at the MS detector were somewhat lower, possibly due to space charge effects in the octapole ion beam guide following the ion funnel. The improved ion transmission properties result primarily from the use of reduced spacings between ring electrodes. We also show that the ion funnel can be operated in two different modes, one using low-rf-amplitude scans, allowing fragile noncovalent complexes (as well as generally undesired adducts) to be transmitted, and the other using high-rf-amplitude scans, providing greater collisional activation and more effective adduct removal (or the dissociation of lower m/z species). PMID- 10845371 TI - Theoretical maximal precision for mass-to-charge ratio, amplitude, and width measurements in ion-counting mass analyzers AB - A theory previously developed for spectra with detector-limited (i.e., signal independent) Gaussian-distributed noise is applied to calculate the maximal precision with which mass spectral peak parameters (mass-to-charge ratio, amplitude, width) can be determined from a discrete spectrum with source-limited Poisson-distributed noise. The precision depends in a calculable way upon the peak shape, signal-to-noise ratio, and number of data points per peak width. Those dependencies are tested by analysis of simulated data. The theory provides estimates for the precision of a repeated experiment, based on data from a single discrete mass spectrum whose parameters are extracted by a least-squares fit to a specified line shape. The relevance of the predictions to present and potential time-of-flight performance is discussed. PMID- 10845372 TI - Chlorine stable isotopes: a comparison of dual inlet and thermal ionization mass spectrometric measurements AB - Chlorine stable isotope ratios, 37Cl/35Cl, currently are measured using dual inlet and thermal-ionization mass spectrometry. These two different analytical techniques, however, have never been cross calibrated. A set of samples with chlorine stable isotope delta values ranging from -4.4 to +0.3 % relative to standard mean ocean water chloride has been analyzed using both of these techniques. Our data show that both techniques can yield similar results within analytical uncertainty. CsCl thermal ionization data are extremely sensitive to the amount of chlorine being measured and cannot be used to determine absolute ratios without an independent means of correcting for machine-induced mass fractionation. As long as standards and samples are of equivalent size, however, the differences between samples measured by thermal ionization remain constant Dual inlet stable isotope mass spectrometry is suited best for samples of > 10 micromol Cl, yielding chlorine stable isotope data with < or =0.1% reproducibilities (2sigma). Thermal ionization mass spectrometry easily accommodates samples of approximately0.1-0.3 micromol Cl, with achievable uncertainties of < or =0.2% (2sigma). PMID- 10845373 TI - Evaluation of electrospray transport CID for the generation of searchable libraries. AB - This paper reports the preliminary investigation of performance-based standard conditions that have been developed for electrospray ionization mass spectrometry. Using performance-based standard criteria, reproducible spectra can be obtained by CID in the electrospray-transport region and searched using a database created using the same performance criteria. To generate library searchable mass spectra, the instrument was tuned to standard conditions that correspond to low, mid, and high fragmentation energies. The instrument was tuned using ion ratios relative to a given peak in a tune compound for each energy level. The library was evaluated using a set of 22 benzodiazepines. The CID libraries were found to be reproducible, both on the same instrument and on different instruments of the same type. Also, the libraries were found to be independent of flow rate and solvent system. The library was expanded to include 16 sulfonylurea herbicides and tested using spiked water samples. Performance of the library was tested over a concentration range of 2 orders of magnitude using sulfonylurea standards. PMID- 10845374 TI - Zeptomole-sensitivity electrospray ionization--Fourier transform ion cyclotron resonance mass spectrometry of proteins. AB - Methods are being developed for ultrasensitive protein characterization based upon electrospray ionization (ESI) with Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS). The sensitivity of a FTICR mass spectrometer equipped with an ESI source depends on the overall ion transmission, which combines the probability of ionization, transmission efficiency, and ion trapping in the FTICR cell. Our developments implemented in a 3.5 tesla FTICR mass spectrometer include introduction and optimization of a newly designed electrodynamic ion funnel in the ESI interface, improving the ion beam characteristics in a quadrupole-electrostatic ion guide interface, and modification of the electrostatic ion guide. These developments provide a detection limit of approximately 30 zmol (approximately 18,000 molecules) for proteins with molecular weights ranging from 8 to 20 kDa. PMID- 10845375 TI - Simultaneous separation of cationic and anionic proteins using zwitterionic surfactants in capillary electrophoresis. AB - The zwitterionic surfactant Rewoteric AM CAS U forms a dynamic wall coating that prevents the adsorption of cationic proteins as well as suppresses the electroosmotic flow (EOF). Addition of polarizable anions to buffers containing this zwitterionic surfactant increases the once suppressed EOF to values nearing +3 x 10(-4) cm2/(V s). The retention of the EOF allows for the separation of analytes of widely different mobilities and is demonstrated by the simultaneous separation of cationic and anionic proteins. Using a buffer containing optimal amounts of the polarizable anion perchlorate and surfactant CAS U, the proteins lysozyme, ribonuclease A, alpha-chymotrypsinogen A, and myoglobin are separated in less than 15 min. Efficiencies as high as 1.5 million plates/m and recoveries greater than 91% are observed for proteins injected in distilled water. Migration time reproducibility is approximately 1% RSD within 1 day and approximately 3% RSD from day to day. The anionic and cationic proteins can be separated over a pH range of 5.5-9, all yielding good efficiencies. PMID- 10845376 TI - Electroosmotically induced hydraulic pumping on microchips: differential ion transport AB - The theory behind and operation of an electroosmotically induced hydraulic pump for microfluidic devices is reported. This microchip functional element consists of a tee intersection with one inlet channel and two outlet channels. The inlet channel is maintained at high voltage while one outlet channel is kept at ground and the other channel has no electric potential applied. A pressure-induced flow of buffer is created in both outlet channels of the tee by reducing electroosmosis in the ground channel relative to that of the inlet channel. Spatially selective reduction of electroosmosis is accomplished by coating the walls of the ground channel with a viscous polymer. The pump is shown to differentially transport ions down the two outlet channels. This ion discrimination ability of the pump is examined as a function of an analyte's electrophoretic velocity. In addition, we demonstrate that an anion can be rejected from the ground channel and made to flow only into the field-free channel if the electrophoretic velocity of the anion is greater than the pressure generated flow in the ground channel. The velocity threshold at which anion rejection occurs can be selectively tuned by changing the flow resistance in the field-free channel relative to the ground channel. PMID- 10845377 TI - Electroosmotic and pressure-driven flow in open and packed capillaries: velocity distributions and fluid dispersion AB - The flow field dynamics in open and packed segments of capillary columns has been studied by a direct motion encoding of the fluid molecules using pulsed magnetic field gradient nuclear magnetic resonance. This noninvasive method operates within a time window that allows a quantitative discrimination of electroosmotic against pressure-driven flow behavior. The inherent axial fluid flow field dispersion and characteristic length scales of either transport mode are addressed, and the results demonstrate a significant performance advantage of an electrokinetically driven mobile phase in both open-tubular and packed-bed geometries. In contrast to the parabolic velocity profile and its impact on axial dispersion characterizing laminar flow through an open cylindrical capillary, a pluglike velocity distribution of the electroosmotic flow field is revealed in capillary electrophoresis. Here, the variance of the radially averaged, axial displacement probability distributions is quantitatively explained by longitudinal molecular diffusion at the actual buffer temperature, while for Poiseuille flow, the preasymptotic regime to Taylor-Aris dispersion can be shown. Compared to creeping laminar flow through a packed bed, the increased efficiency observed in capillary electrochromatography is related to the superior characteristics of the electroosmotic flow profile over any length scale in the interstitial pore space and to the origin, spatial dimension, and hydrodynamics of the stagnant fluid on the support particles' external surface. Using the Knox equation to analyze the axial plate height data, an eddy dispersion term smaller by a factor of almost 2.5 than in capillary high-performance liquid chromatography is revealed for the electroosmotic flow field in the same column. PMID- 10845378 TI - A multicomponent mobile phase for ion chromatography applied to the separation of anions from the residue of low explosives AB - A multicomponent mobile phase utilizing ion-exchange, ion-exclusion, and ion pairing principles for the rapid isocratic separation of anions in low explosives residue by ion chromatography (IC) has been developed. The notable feature of this system is that an ion-pairing reagent and an ion-exclusion reagent are combined in the same mobile phase. Contrary to expectation, these reagents act independently of each other in solution. The stock mobile-phase composition consisted of boric acid, D-gluconic acid, lithium hydroxide, and glycerol. Tetrapropylammonium hydroxide, an ion-interaction reagent was used to achieve pH 8.5. Acetonitrile (ACN) was added to enhance resolution and octanesulfonic acid, an ion-exclusion reagent, was added to adjust the retention time of perchlorate. Separation of a mixture of anions common to low explosives residue was achieved in less than 16 min using a Waters IC-Pak Anion HR column. Optimization studies were performed by changing the concentration of the ACN and by altering the pH or the type of ion-interaction or -exclusion agents. Simulated case studies were performed using postblast residues from pipe bombs. The results show this method to be a valid and reproducible procedure for forensic casework analysis. The practical significance of this system is that a reduction in the analysis time and an improvement in efficiency of late-eluting peaks can be achieved without resorting to gradient elution techniques. For the analysis of anions detected in explosives residue, the Waters IC-Pak Anion HR column has proven to be a suitable replacement for the Vydac 300IC405 column, which has been discontinued by the manufacturer. PMID- 10845379 TI - Microextraction of nine haloacetic acids in drinking water at microgram per liter levels with electrospray-mass spectrometry of stable association complexes AB - Haloacetic acids are disinfection byproducts of the chlorination of drinking water. This paper presents the analysis of all nine chloro and bromohaloacetic acids (HAA9) at the submicrogram per liter level by microextraction with detection by electrospray-mass spectrometry. The haloacetic acids are extracted from acidified water through a microscale liquid-liquid extraction. Perfluoroheptanoic acid is added to the extracts, and the haloacetic acids are detected with electrospray-mass spectrometry. Confidence in the selective quantification of the haloacetic acids is achieved by observing the stable association complexes that are formed between the haloacetic acids and perfluoroheptanoic acid. The method detection limits for the haloacetic acids are less than 1 microg L(-1), depending on the haloacetic acid. Standard addition is used to quantify the haloacetic acids in several water matrixes. PMID- 10845380 TI - Use of solid-phase microextraction for the quantitative determination of herbicides in soil and water samples. AB - An in-depth study of SPME optimization and application has been made, considering not only aqueous (surface water and groundwater samples) but also the more complex soil samples. Seven herbicides widely used in the area of study have been selected including five triazine herbicides (atrazine, simazine, terbumeton, terbuthylazine, terbutryn), molinate, and bromacil. linearity range was between 0.1 and 10 ng/mL and the repeatability below 10% when applying the optimized SPME procedure to water samples. Reproducibility was found to be lower than 20% at the 1 ng/mL level, and the limits of determination in environmental water samples using GC/MS (SIM mode) were well below 0.1 ng/mL (values ranging from 10 to 60 ng/L). Extraction of selected herbicides from soil was carried out by microwave assisted solvent extraction using methanol in screw-capped vials, leading to recoveries over 80% in spiked soil samples at the 5-200 ng/g level. SPME application over methanolic soil extracts required a 10-fold dilution with distilled water. The recommended procedure was found to be fully applicable for quantitative determination of selected herbicides in soils containing low organic matter content with coefficients of variation below or around 10% and limits of determination ranging from 1 to 10 ng/g. Both procedures were applied to real world surface water and soil samples where several pesticides were detected including atrazine, simazine, terbuthylazine, and molinate. PMID- 10845381 TI - An on-line HPLC method for detection of radical scavenging compounds in complex mixtures. AB - A rapid on-line method for screening of complex mixtures for radical scavenging components was developed using a methanolic solution of 2,2'-diphenyl-1 picrylhydrazyl (DPPH) stable free radical. The HPLC-separated analytes react postcolumn with the DPPH solution, and the induced bleaching is detected as a negative peak by an absorbance detector at 517 nm. An optimized instrumental setup is presented. The method is suitable for both isocratic and gradient HPLC runs with mobile-phase compositions ranging from 10 to 90% organic solvent in water or buffer (pH 3-6). The method is simple, has a broad applicability, and uses common instruments, inexpensive and stable reagents, and a time-saving and nonlaborious experimental protocol. It can also be used for quantitative analysis. The method was applied to several pure natural antioxidants and plant extracts. The limits of detection were 0.33-94 microg/mL, depending on the compound tested. PMID- 10845382 TI - Financial viability, medical technology, and hospital closures. AB - Informed investments in medical technology and information systems are associated with the financial viability of community hospitals. Financially distressed facilities are 3 to 4 years behind proactive hospitals in supporting high-speed data, voice, and image transmissions to physicians in various locations. Impact of the Balanced Budget Act of 1997, fraud and abuse activities, Y2K issues, and lack of information systems support for physicians will result in 800 hospital closures and mergers of distressed hospitals over the next 60 months. These findings are based on the application of an eight-step framework for classifying information systems in health care entities. This framework is validated by survey instruments, site visits, interviews with senior management in 44 health care entities containing 576 hospitals, and judgments on the financial status of the health care entities. PMID- 10845383 TI - Financial management in leading health care systems. AB - To understand better the financial management practices and strategies of modern health care organizations, we conducted interviews with chief financial officers (CFOs) of several leading health care systems. In this introduction, we present an overview of the project and summary responses on corporate financial structures and strategic challenges facing CFOs. PMID- 10845384 TI - Capital investment strategies in health care systems. AB - Capital investment decisions are among the most important decisions made by firms. They determine the firm's capacity for providing services and commit the firm's cash for an extended period of time. Interviews with chief financial officers of leading health care systems reveal capital investment strategies that generally follow the recommendations of modern finance theory. Still, there is substantial variation in capital budgeting techniques, methods of risk adjustment, and the importance of qualitative considerations in investment decision making. There is also variation in delegation of investment decision making to operating units and methods of performance evaluation. Health care systems face the same challenges as other organizations in developing and implementing capital investment strategies that use consistent methods for evaluation of projects that have inconsistent aims and outcomes. PMID- 10845385 TI - Capital structure strategy in health care systems. AB - The capital structures (the relative use of debt and equity to support assets) of leading health care systems are viewed as a strategic component of their financial plans. While not-for-profit hospitals as a group have maintained nearly constant levels of debt over the past decade, investor-owned hospitals and a group of leading health care systems have reduced their relative use of debt. Chief financial officers indicated that in addition to reducing debt because of less favorable reimbursement incentives, there was a focus on maintaining high bond ratings. Debt levels have not been reduced as sharply in these health care systems as they have in investor-owned hospitals, in part due to the use of debt to support investments in financial markets. Because these health care systems do not have easy access to equity, high bond ratings and solid investment earnings are central to their capital structure policies of preserving access to debt markets. PMID- 10845386 TI - Sources of project financing in health care systems. AB - Through discussions with chief financial officers of leading health care systems, insights are offered on preferences for project financing and development efforts. Data from these same systems provide at least anecdotal evidence in support of pecking-order theory. PMID- 10845387 TI - Cash management in health care systems. AB - Cash is one of the most precious assets held by health systems. This article presents results of discussions on cash management and investment policies at a dozen major not-for-profit health systems. Health system data indicate that cash holdings have increased dramatically since 1993, mostly due to investment earnings. Discussions with chief financial officers of these health systems reveal that cash holdings are significant, that decisions about cash balances are strategic, and that most systems aim to increase cash balances to levels that permit access to capital markets on more favorable terms. PMID- 10845388 TI - Improved inpatient billing metrics by physician-directed billing software. AB - Billing may not be prominent on academic physicians' lists of priorities because of other demands and commitments for their time. The authors developed and examined the use of an electronic inpatient billing tool that also facilitated physician rounding responsibilities. The results show that use of an electronic tool was more acceptable to physicians than their original card system, and it significantly decreased charge-processing lag time. PMID- 10845389 TI - Segmenting health maintenance organizations to study productivity and profitability. AB - As the decade ended, health maintenance organizations (HMOs) were increasing in popularity as a means of health care delivery. These groups take many forms, so it is important for the analyst to see if the efficiency and financial results for these different forms vary. The four major forms are profit vs. not-for profit, chain vs. non-chain, group/staff vs. individual practice association (IPA), and federally qualified vs. non-federally qualified. Using a nationwide database of all the HMOs in the United States, the article compares liquidity rates, leverage ratios, profitability ratios, marketing, and per member ratios across the four groups using paired t tests. The two classifications that showed the most differences were group/staff vs. IPA and federally qualified vs. non federally qualified. IPAs have a better liquidity position and lower leverage ratios than group/staff but their administrative costs are higher and the time to receive payments and to pay debts is higher. Non-federally qualified have somewhat higher liquidity ratios and higher profitability ratios. These significant differences in financial outcomes indicate that studies of HMOs should segment different major forms of organizations and study them separately before trying to show the effects of different policies on HMO efficiency and effectiveness. PMID- 10845390 TI - Prognostic value of proliferation index and expression of the RNA component of human telomerase (hTR) in papillary meningiomas. AB - Papillary meningioma is a rare subtype of meningioma that often behaves aggressively. In order to characterize factors that may influence this behavior, we chose to compare MIB-1 labeling index (LI) and telomerase RNA localization (hTR) in papillary meningiomas, meningiomas, and atypical meningiomas. LI is now often used to supplement histologic grade in the evaluation of these lesions. More recent studies indicate that increased expression of hTR is detected in many neoplastic cells, and may play an essential role in cell immortalization. The study group consisted of five papillary meningiomas (and a recurrence in one case), 11 conventional meningiomas, and eight atypical meningiomas. Conventional meningiomas showed either negative or 1 + hTR. Atypical meningiomas showed 1 + hTR. Papillary meningiomas showed the highest hTR (five of six, including recurrence, 2-3+ and one 1+). Generally, the LI was very low for conventional meningiomas (< 2%). The LI of atypical meningiomas ranged from 3-19%, mean 12%, and from 5.5-17.5%, mean 11.75% for papillary meningiomas. LI differentiated between meningiomas, and papillary or atypical meningiomas. hTR further delineated papillary (moderate to high) from atypical meningiomas (low). The combined variable of LI and hTR expression could be a useful independent prognostic indicator in patients with papillary meningioma. PMID- 10845391 TI - Interleukin-6 transduction of a rat T9 glioma clone results in attenuated tumorigenicity and induces glioma immunity in Fischer F344 rats. AB - We transduced a highly tumorigenic T9 clone (T9.F), isolated from the rat T9 glioblastoma cell line, with a retroviral expression vector containing the human IL-6 cDNA and investigated the effects of IL-6 secretion on glioma formation in the syngeneic Fischer rat. Two subclones producing high and low levels (35 and 3.5 ng/10(6) cells/48 h) of IL-6 were identified and were termed T9.F/IL6/hi and T9.F/IL6/lo, respectively. Subcutaneous (SC) injection of 1 x 10(6) parental T9.F cells resulted in 100% tumor formation and progression. When 1 x 10(6) IL-6 secreting T9.F cells were injected SC, a small palpable tumor formed which sometimes regressed. In this regard, no tumors were detected after 30 days in 76% (13/17) of animals injected with T9.F/IL6/hi cells, whereas only 10% (1/10) of the rats injected with T9.F/IL6/lo cells completely rejected their tumors within this time frame. The addition of an IL-6 neutralizing antibody to the T9.F/IL6/hi SC inoculum followed by an intratumoral injection of the IL-6 neutralizing antibody, seven days later, abrogated the anti-tumor effects. Animals that rejected the IL-6 secreting tumors were 100% protected from subsequent intracranial (IC) challenges with the parental T9.F glioma as well as the original T9 glioblastoma; partially protected from an IC challenge with the unrelated, syngeneic RT-2 glioma; but were not protected from an IC challenge with the syngeneic MadB106 adenocarcinoma. When 1 x 10(4) cells were injected in the brain of naive animals, survival time was significantly increased for those rats implanted with T9.F/IL6/hi cells, but not T9.F/IL6/lo cells, as compared to animals implanted with T9.F parental cells (p = 0.003). This study demonstrates that IL-6 secretion attenuates SC and IC glioma growth and SC rejection of IL-6 secreting T9.F cells induces long-term glioma immunity which is effective in the brain. PMID- 10845392 TI - Prognostic value of immunoexpression of the chemoresistance-related proteins in ependymomas: an analysis of 76 cases. AB - Intracranial ependymomas are the third most common primary brain tumor in children. A variety of chemotherapy protocols have been introduced for the treatment of ependymoma although overall these have not contributed to patients outcome. To our knowledge, data on the prognostic value of immunoexpression of the chemoresistance-related proteins (ChRPs) in ependymomas are absent. Seventy six patients with intracranial ependymomas who received combined treatment were studied retrospectively. Tumor specimens were immunohistochemically examined with antibodies to metallothioneins (MT), glutathione S-transferase pi (GST pi) and P glycoprotein (P-GP). The results demonstrated significant preponderance of expression of all the above-mentioned ChRPs for the low-grade tumors. The progression-free survival time was found to be significantly shorter for immunonegative tumors in both tumor grades. Multivariate analysis using a Cox hazard model revealed that recurrence-free survival time is significantly associated with tumor grade, and MT and P-GP expression. Risk of recurrence increased for the high-grade ependymomas (hazard ratio 2.85; P = 0.004), and decreased for the MT-positive tumors (hazard ratio -2.72; P = 0.005) and for the P-GP-positive tumors (hazard ratio -2.02; P = 0.02). The obtained results allow one to conclude that ChRPs expression is closely associated with low-grade ependymomas and immunohistochemical findings may be estimated as a predictor for local tumor progression. PMID- 10845393 TI - Treatment of supratentorial glioblastoma multiforme with radiotherapy and a combination of BCNU and tamoxifen: a phase II study. AB - From May 1990 to November 1994, 70 consecutive patients suffering from glioblastoma multiforme were treated following surgery with conventional radiotherapy and adjuvant IV BCNU administered alone or in combination with tamoxifen. Twenty-five patients received BCNU alone (control group A) while 24 patients also received 40 mg of tamoxifen (TMX) PO daily (group B) and 21 received 100 mg of TMX PO daily (group C). There were no significant differences between the 3 groups concerning age, type of resection and median post-operative Karnofsky performance status (KPS). Blood toxicity over grade II occurred in 33.5% of patients receiving TMX versus 12% of patients treated with BCNU alone (p < 0.05). Deep venous thrombosis complications were observed in 4 patients of each TMX group, whereas they were not observed in the control group (p < 0.04). Median time to tumor progression (MTTP) was 35 weeks in the control group and 27 weeks in both TMX groups B and C. Median survival time (MST) was 56, 66 and 51 weeks, respectively. These results suggest that the addition of TMX to standard treatment of glioblastomas does not affect the time to tumor progression and overall survival but may increase the risk of deep venous thrombosis or nitrosourea-induced blood toxicity. PMID- 10845394 TI - Choroid plexus metastasis from carcinoma of the bladder: case report and review of the literature. AB - We present here a case report of a 40-year old male with adenocarcinoma of the bladder and solitary metastasis to the choroid plexus of the right lateral ventricle. This is the first such report of such a metastasis in association with bladder carcinoma. Systemic metastases frequently occur in patients with carcinoma of the bladder but involvement of central nervous system is relatively uncommon: less than 1% of patients with carcinoma of the bladder present an intracerebral metastasis. In the majority of cases there are either multiple CNS metastases or other distant metastases. A few cases present with solitary metastases to the CNS without evidence of recurrent or disseminated disease. PMID- 10845395 TI - Peripheral neuropathy due to biweekly paclitaxel, epirubicin and cisplatin in patients with advanced ovarian cancer. AB - We assessed the peripheral neuropathic changes induced by biweekly combination chemotherapy including paclitaxel 100-165 mg/m2 (in a 3-h infusion), epirubicin 75 mg/m2 and cisplatin 50 mg/m2 (TEC) in patients with advanced ovarian cancer. Neurologic evaluation, including a standardized questionnaire, bed-side neurological examination, and quantitative determination of vibratory perception thresholds (VPT) and grip strength took place before therapy, after 3 and 6 cycles, and thereafter whenever possible. During chemotherapy all patients received granulocyte colony-stimulating factor from days 2 to 12. Pretreated patients received amifostine two times, before epirubicin and before cisplatin administration. Neuropathic symptoms developed in 11/13 non-pretreated patients and in 7/9 chemotherapy-pretreated patients. Neuropathic signs developed in all patients. Neuropathic symptoms and signs were predominantly sensory in character. VPT changes developed primarily in the feet. According to National Cancer Institute of Canada Common Toxicity Criteria, grade 3 peripheral neuropathy after 6 cycles developed in 1/6 and 2/4 non-pretreated patients who received TEC containing paclitaxel 150 and 165 mg/m2, respectively. We conclude that peripheral neuropathy is dose-limiting in chemonaive patients treated with biweekly TEC combination chemotherapy, at paclitaxel dose level 165 mg/m2 in a 3 h intravenous administration. PMID- 10845397 TI - The problem resident. PMID- 10845396 TI - Antiangiogenic therapy in brain tumor models. AB - The prognosis of patients with malignant brain tumors remains poor despite new developments in neurosurgery, chemotherapy and radiotherapy. Malignant gliomas are highly vascularized, and there is ample evidence that their growth is angiogenesis-dependent. Therefore, new therapeutic approaches often include the inhibition of angiogenesis. In this review, experimental studies of antiangiogenic agents in brain tumor models are summarized. The results of these experiments as well as potential pitfalls in extrapolation to the clinic are discussed. PMID- 10845398 TI - Computer-aided, case-based diagnosis of mammographic regions of interest containing microcalcifications. AB - RATIONALE AND OBJECTIVES: The purpose of this study was to evaluate the effectiveness of a mapped-database diagnostic system in reducing the incidence of benign biopsies and misdiagnosed cancers among mammographic regions of interest (ROIs). MATERIALS AND METHODS: A novel neural network was devised (a) to respond to a query ROI by recommending to biopsy or not to biopsy and (b) to map each ROI in the database as a dot on a computer screen. The network was designed so that clusters in the array of dots help the radiologist to find proved ROIs visually similar to the query ROI. This mapped-database diagnostic system was restricted to ROIs with visible microcalcifications. The neural network was trained with a stored database of 80 biopsy-proved ROIs. RESULTS: Four radiologists acting independently on 100 ROIs recommended biopsies for 18, 15, 28, and 18 benign ROIs and misdiagnosed cancers in 11, 12, 7, and eight ROIs, respectively. Interaction with the mapped-database system reduced the numbers of benign biopsies to 11, eight, 18, and 10 cases and of misdiagnosed cancers to eight, seven, four, and three cases, respectively. Statistical analysis indicated that three radiologists achieved significant improvements at P < or = .02 and the fourth achieved a substantial improvement at P < or = .07. CONCLUSION: By using a mapped database of proved mammographic ROIs containing microcalcifications, radiologists may statistically significantly reduce the numbers of benign biopsies and misdiagnosed cancers. PMID- 10845399 TI - Optimizing parameters for computer-aided diagnosis of microcalcifications at mammography. AB - RATIONALE AND OBJECTIVES: The purpose of this study was to optimize selection of the mammographic features most useful in discriminating benign from malignant clustered microcalcifications. MATERIALS AND METHODS: The computer-aided diagnosis (CAD) system automatically extracted from digitized mammograms 13 quantitative features characterizing microcalcification clusters. Archival cases (n = 134; patient age range, 31-77 years; mean age, 56.8 years) with known histopathologic results (79 malignant, 55 benign) were selected. Three radiologists at three facilities independently analyzed the microcalcifications by using the CAD system. Stepwise discriminant analysis selected the features best discriminating benign from malignant microcalcifications. A classification scheme was constructed on the basis of these optimized features, and its performance was evaluated by using receiver operating characteristic (ROC) analysis. RESULTS: Six of the 13 variables extracted by the CAD system were selected by stepwise determinant analysis for generating the classification scheme, which yielded an ROC curve with an area (Az) of 0.98, specificity of 83.64%, positive predictive value of 89.53%, and accuracy of 91.79% for 98% sensitivity. When patient age was an additional variable, the scheme's performance improved, but this was not statistically significant (Az = 0.98). The ROC curve of the classifier (without age as an additional variable) yielded a high Az of 0.96 for patients younger than 50 years and an even higher (P < .02) Az of 0.99 for those 50 years or older. CONCLUSION: Stepwise discriminant analysis optimized performance of a classification scheme for microcalcifications by selecting six optimized features. Scheme performance was significantly (P < .02) higher for women 50 years or older, but the addition of patient age as a variable did not produce a statistically significant increase in performance. PMID- 10845400 TI - Bootstrap estimation of diagnostic accuracy with patient-clustered data. AB - RATIONALE AND OBJECTIVES: The purpose of this study was to describe a simple bootstrap approach for estimating sensitivity, specificity, and the area under the receiver operating characteristic curve for multisite test outcome data. MATERIALS AND METHODS: The performance of bootstrap estimates was evaluated and compared with that of analytic estimates by using a simulation study. Bootstrapping was demonstrated by using data from a previous study comparing two angiographic methods. RESULTS: Analytic and bootstrap estimates had similar coverage rates for 95% confidence intervals. With many sites per patient, bootstrap estimates had slightly better coverage than analytic estimates. Bootstrap percentile intervals had better coverage than asymptotic normal bootstrap intervals. CONCLUSION: Bootstrapping is a useful method of estimating confidence intervals for the area under the receiver operating characteristic curve, sensitivity, and specificity when data are correlated. PMID- 10845401 TI - A contaminated binormal model for ROC data: Part I. Some interesting examples of binormal degeneracy. AB - RATIONALE AND OBJECTIVES: Receiver operating characteristic (ROC) data with false positive fractions of 0 are often difficult to fit with standard ROC methods and are sometimes discarded. Some extreme examples of such data were analyzed to evaluate the nature of these difficulties. MATERIALS AND METHODS: Rating reports of fracture for single-view ankle radiographs were analyzed with the binormal ROC model and with two ROC models that keep the ROC curve from crossing the chance line. Because fractures were almost never reported that were not present, some views and locations yielded only ROC points with false-positive fractions of 0, while others yielded at least one ROC point with a non-0 false-positive fraction. RESULTS: The models tended to yield ROC areas close to or equal to 1. ROC areas of 1 imply a true-positive fraction close to 1; yet the data contained no such fractions. When all false-positive fractions were 0, the true-positive fraction could be much higher for one view than another for all observers. ROC areas gave little or no hint of these unmistakable differences in performance. CONCLUSION: These data challenge the validity and robustness of current ROC models. A key aspect of ankle fractures is that some may be visible on one view but not at all visible on another. PMID- 10845402 TI - A contaminated binormal model for ROC data: Part II. A formal model. AB - RATIONALE AND OBJECTIVES: A contaminated binormal receiver operating characteristic (ROC) model is proposed to account for ROC data that have very few false-positive reports even though many healthy subjects are sampled. The model assumes that no signal information is captured for a proportion of abnormalities, and that these abnormalities have the same distribution as noise along the latent decision axis. MATERIALS AND METHODS: The authors developed a formal psychophysical model, presented here in detail. They have specified the psychophysical assumptions of the theory, and have provided proofs that include all essential details, from assumptions to implications. With the technical details that are provided, this theory can be implemented with computer programs to fit data. RESULTS: The new model can fit ROC data in which some or all of the ROC points have false-positive fractions of 0 and true-positive fractions of less than 1, without implying that performance is perfect. The resulting ROC curves are always proper, never exhibiting inappropriate chance line crossings. The model predicts that, under certain conditions, a bimodal categorical rating histogram will be observed for the signal distribution. The model predicts a relationship between the mean and standard deviation of the signal distribution and holds that, for expert decision makers, there are situations in which the prevalence and utility matrix preclude operating points in some ROC regions. The model has a straightforward extension to the joint detection and localization ROC curve. CONCLUSION: The contaminated binormal model accounts for ROC data with few or no false-positive reports. PMID- 10845403 TI - A contaminated binormal model for ROC data: Part III. Initial evaluation with detection ROC data. AB - RATIONALE AND OBJECTIVES: The authors' purpose was to evaluate how well the contaminated binormal receiver operating characteristic (ROC) model fits (a) degenerate data for which standard ROC models commonly fail and (b) nondegenerate data from exemplary experiments, for which the standard binormal model should be appropriate. MATERIALS AND METHODS: The authors studied two examples of binormally degenerate data, with and without interior points, and ROC rating data from four experiments in visual psychophysics and radiology. The plots of contaminated binormal ROC curves of the binormal degenerate data were examined. For ROC data with at least one interior point, the new model was compared with conventional models on the basis of likelihood-ratio chi2 statistics (G2). RESULTS: With no interior points, the contaminated binormal model gave results consistent with the fundamental principle underlying ROC analysis, that is, for a fixed false-positive probability, the higher the true-positive probability, the better the diagnostic performance. Contaminated binormal ROC curves go through the empirical ROC points of the degenerate data without crossing the chance line or climbing far above the true-positive fractions of the points. For several model ROC studies, the contaminated binormal model gave smaller G2 results than conventional ROC models, although the differences tended to be small, usually with little difference in ROC area. CONCLUSION: The contaminated binormal model fits binormal degenerate data better than conventional ROC models, and it offers an explanation for the degeneracy. The lower G2 values on some classic, nondegenerate ROC data suggest that contamination may not be limited to degenerate ROC data. PMID- 10845404 TI - The problem resident: the perspective of chief residents. PMID- 10845405 TI - Personal diary: a patient's experience at an academic radiology department. PMID- 10845406 TI - Charles A. Rohrmann, Jr, MD: 2000-2001 Armed Forces Institute of Pathology Distinguished Scientist. PMID- 10845407 TI - National Cancer Institute initiative for development of novel imaging technologies. PMID- 10845408 TI - Ambulatory use of parenteral antibacterials: contemporary perspectives. AB - Outpatient parenteral antimicrobial therapy (OPAT) offers increased patient comfort and convenience in appropriately selected patients who require parenteral antibacterial therapy, as well as opportunity for cost savings. Home-based programmes, with drugs being administered by the patient or the caregiver, have become the norm in the USA. Choice of drugs for OPAT is based on antimicrobial spectrum, dosage regimen, drug stability, toxicity profile, and cost. Over the past decade, availability of sophisticated programmable pumps has allowed a wider range of antimicrobial agents to be used in the ambulatory setting. The most popular antibacterial agents in OPAT programmes in the USA are vancomycin and beta-lactams. PMID- 10845410 TI - Parenteral cephalosporin therapy in ambulatory care: advantages and disadvantages. AB - Outpatient parenteral antibiotic therapy (OPAT) programmes are effective, well tolerated and economically advantageous in carefully selected patient populations. Inclusion criteria for patient selection for OPAT include good clinical appearance and uncomplicated infection. By virtue of their favourable microbiological and pharmacological properties, cephalosporins in general, and ceftriaxone in particular, are the most widely prescribed antibiotics for OPAT worldwide. OPAT was largely created to prolong parenteral therapy following early discharge and has now been extended to community general practice. Indeed, more than 250000 treatments are performed in the US each year for a wide variety of serious infections, with an increase of 100% during the last 5 years. In 1994, an advisory committee was created in Canada to provide guidelines for home intravenous therapy. Of the 3 models that were defined (the visiting nurse model, the infusion centre model and the self-administration model), the OPAT self administration model offers considerable cost savings and is probably largely utilised in a number of countries, such as Italy, where specific models have not been codified. Once the need for parenteral antibiotic therapy has been established, the choice of antibiotic is the second step in the decision-making process. Third generation cephalosporins are characterised by a number of important advantages in the OPAT setting, namely a favourable antibacterial spectrum, tolerability profile and patient compliance, as well as advantageous cost considerations. While the advantages of parenteral cephalosporin therapy in the ambulatory care setting outweigh the disadvantages in terms of cost effectiveness and rapid onset of action, adverse events such as pain at the injection site following intramuscular administration and phlebitis after intravenous infusion should be borne in mind. PMID- 10845409 TI - Pharmacological properties of parenteral cephalosporins: rationale for ambulatory use. AB - Parenteral cephalosporins are among the most frequently used antibiotics in hospital therapy. They are characterised by an extended spectrum of activity against gram-positive and gram-negative bacteria, and some also have good activity against anaerobes. They kill proliferating bacterial cells rapidly, and generally show only a low tendency to select resistant mutants. However, there are cephalosporin compounds which induce cephalosporinases very rapidly in certain microorganisms. Together with other beta-lactam antibiotics, parenteral cephalosporins interfere with bacterial cell wall synthesis by inhibiting peptidoglycan cross-linkage. Because of this specific target, they are nontoxic to mammalian cells, and have a very favourable adverse effect profile. The chemical stability of parenteral cephalosporins in aqueous solution is good. After intravenous injection, high concentrations of these agents are achieved in serum and tissue. Most cephalosporins are eliminated unchanged via the kidney, with a half-life of 1 to 2 hours. But there are also derivatives with a serum half-life of more than 2 and up to 8 hours, allowing 12- or 24-hour dosage intervals. Because of their reliable efficacy and low risk of adverse effects, the parenteral cephalosporins offer a high degree of tolerability even in the setting of outpatient antibiotic therapy. In particular, the derivatives of the third generation are characterised by unique pharmacological properties. PMID- 10845411 TI - Pharmacoeconomic considerations in the ambulatory use of parenteral cephalosporins. AB - It has been clearly documented that outpatient parenteral antibiotic therapy (OPAT) saves money compared with hospital care for patients who need intravenous antimicrobial therapy. The reduced expenses come primarily from savings in facility and hospital staffing costs. In addition to shortening hospital stay, OPAT programmes can be developed so that hospital care is avoided altogether. However, even with the clear potential for savings, to have a successful programme it is necessary to align the interests of the payers, the physicians, the administrators and the patients. The cost of OPAT programmes can also be reduced through patient evaluation and careful selection of the appropriate delivery model, antibiotic, dosage intervals and infusion technology. The fact that antibiotics such as ceftriaxone, the aminoglycosides and vancomycin can be given once daily in the elderly offers particular advantages in terms of convenience as well as cost. In order to achieve cost savings, managed care will increasingly rely on home and outpatient therapy. This pressure will need to be counterbalanced by quality assurance programmes and outcomes measurements. PMID- 10845413 TI - Prognostic factors in retinoblastoma. PMID- 10845414 TI - New developments in neonatology: less severe retinopathy of prematurity? AB - PURPOSE: To determine the effects of surfactant replacement therapy (SRT), high frequency oscillatory ventilation (HFOV), and general improvements in quality of care on the incidence of severe retinopathy of prematurity (ROP). METHODS: Retrospective comparison of the incidence and severity of ROP in two groups of preterm infants admitted to our neonatal intensive care unit (NICU) in two consecutive 5-year periods (1986-1995) and screened for ROP. During the second study period, natural surfactant was introduced in the treatment of respiratory distress syndrome (RDS) and HFOV was used for treatment of respiratory insufficiency of any origin. The effects of these developments and general improvements on the incidence of severe ROP were analyzed with stepwise logistic regression. RESULTS: The overall incidence of ROP from 1986-1990 was not significantly different from the incidence of ROP from 1991-1995. The incidence of severe ROP (ROP stage 3 or greater) was significantly lower in the second period (15.7% versus 6.4%, P=.015). For infants <1000 g, the incidence of overall ROP was increased significantly during the second study period (47.6% versus 60.1 %, P=.045), although the incidence of severe ROP remained unchanged. Only SRT was associated with a decreased risk for severe ROP; HFOV and general improvements in quality of care had no influence on the outcome. In patients with RDS, the incidence of severe ROP decreased significantly during the second period. CONCLUSION: Of the recent new developments in neonatology, only SRT was associated with a decreased risk for severe ROP. PMID- 10845412 TI - Place of parenteral cephalosporins in the ambulatory setting: clinical evidence. AB - During the last decade, 6 parenteral third generation cephalosporins have been introduced into clinical practice. The three most frequently used agents are cefotaxime, ceftazidime and ceftriaxone. Although primarily used in hospitals, these agents are increasingly employed in the ambulatory setting. In particular, ceftriaxone, because of its favourable pharmacokinetic profile allowing once daily administration by a bolus injection, has demonstrated both tolerability and efficacy in the ambulatory setting during extensive worldwide use. Sophisticated parenteral infusion systems enable cephalosporins that require more frequent administration to be delivered in this setting. In noncomparative studies involving a range of patient populations and serious infections (mostly bone, joint and soft tissue, and pneumonia and febrile episodes in neutropenia), these cephalosporins achieved equivalent efficacy and tolerability, and considerable cost savings, since patients were able to receive all or part of their treatment in the home or outpatient setting. However, more comparative studies of ambulatory parenteral therapy in the inpatient setting or ambulatory oral therapy are necessary to further clarify the true cost effectiveness of this type of healthcare delivery. This is increasingly relevant in countries where parenteral antimicrobials are not the 'standard of care' in managing many serious infections. Published experience to date confirms that third generation cephalosporins, particularly ceftriaxone, should have an essential place in the therapeutic formulary of any ambulatory parenteral antibiotic programme. PMID- 10845416 TI - Effect of bipolar cautery on tensile strength of 6-0 polyglactin 910 suture. AB - PURPOSE: To determine the effect of bipolar cautery on 6-0 polyglactin 910 suture. METHODS: Three groups of sutures were tested for maximum tensile strength. The first group (direct cautery) had bipolar cautery applied directly to the suture. The second group (scleral cautery) had bipolar cautery applied directly on top of the sclera, through which the suture was passed one-half scleral thickness. The third group (control) had no application of cautery to the suture. RESULTS: The mean tensile strength for the direct cautery sutures (n=8) was 553 g; for the scleral cautery sutures (n=7), 577 g; and for the control sutures (n=5), 553 g. There was no statistically significant difference in tensile strength between the three groups, with the analysis of variance yielding P=.56. CONCLUSION: Bipolar cautery causes little to no risk of reducing the tensile strength of 6-0 polyglactin 910 suture in the setting of strabismus surgery. PMID- 10845415 TI - Anterior transposition of the superior oblique tendon in the treatment of oculomotor nerve palsy and its influence on postoperative hypertropia. AB - PURPOSE: To determine whether postoperative hypertropia after anterior transposition of the superior oblique tendon without trochleotomy could be avoided with a simplified surgical approach. METHODS: Eight patients with oculomotor nerve palsy (one patient was bilaterally affected) were retrospectively identified as having undergone anterior transposition of the superior oblique tendon without trochleotomy or vertical rectus muscle surgery between March 1992 and September 1998. The superior oblique tendon was cut at the medial border of the superior rectus muscle and placed 1-3.5 mm anterior to the medial insertion of the superior rectus muscle in each of these patients. Resection of the superior oblique tendon was not performed. The lateral rectus muscle was weakened, and no vertical rectus muscles were resected. RESULTS: Preoperative deviations with the uninvolved eye fixating in primary position ranged from 20-90 prism diopters (delta) of exotropia (mean: 49.3 delta) and from 0-20 delta of hypotropia (mean: 11.25 delta). Postoperative horizontal deviations in the primary gaze position ranged from 12 delta of exotropia to 20 delta of esotropia. Six cases were aligned within 10 delta of exotropia or esotropia. Postoperative vertical deviations in the primary gaze position ranged from 2 delta of hypertropia to 8 delta of hypotropia. Six cases were aligned within 2 delta of deviation. Significant postoperative restrictive hypertropia, or new postoperative paradoxical ocular movements, did not occur in any patient. Patients who underwent follow-up >4 months maintained stable eye alignment. CONCLUSION: Transposition of the superior oblique tendon without simultaneous resection or trochleotomy, or additional surgery to the vertical rectus muscle simplifies the surgical technique and eliminates subjective decision making regarding the amount of resection. PMID- 10845417 TI - Sensory deviations subsequent to senile cataract. AB - PURPOSE: To evaluate the effect of extracapsular cataract extraction with posterior chamber intraocular lens (IOL) implantation in patients with sensory deviations subsequent to senile cataract. METHODS: Twenty patients with dense cataract and associated sensory deviations underwent follow-up between April 1996 and April 1998 after extracapsular cataract extraction with posterior chamber IOL implantation in the deviating eye. All patients underwent follow-up for a minimum period of 6 months. RESULTS: Preoperatively, all patients had a visual acuity of PL positive (perception of light present) and PR (projection of rays) accurate in the deviating eye. Two patients had an esodeviation, and 18 patients had an exodeviation. Postoperatively, all patients had a corrected visual acuity of 20/40 or better in the operated eye at 12 weeks. Nineteen patients had ocular alignment within +/-8 prism diopters of orthophoria at 12 weeks. CONCLUSION: Sensory deviations subsequent to senile cataract usually resolve spontaneously after cataract surgery if visual gains are 20/40 or better. The prognosis for binocular vision is good. PMID- 10845418 TI - Combined effect of polytetrafluoroethylene and 5-fluorouracil on delayed adjustable strabismus surgery. AB - PURPOSE: To determine a more reliable method for performing delayed suture adjustment to prevent the formation of postoperative adhesions and delay the time of adjustment. METHODS: Thirty-two rabbit eyes were divided into three groups. After recession of the superior rectus muscle, 5-fluorouracil (5-FU) was administered beneath and over the superior rectus muscle in group 5-FU (12 eyes); polytetrafluoroethylene (PTFE) and 5-FU were administered in group P-f (10 eyes); and PTFE, 5-FU, and Viscoat (chondroitin sulfate, sodium hyaluronate, Alcon Laboratories Inc, Fort Worth, Tex) were administered in group P-fv (10 eyes). Delayed adjustment was performed once on each superior rectus muscle at 1, 2, and 3 weeks postoperatively in group 5-FU and at 2 and 4 weeks postoperatively in groups P-f and P-fv. The length and force for the adjustment and the degree of adhesions were recorded. RESULTS: The use of fluorouracil allowed adjustment to be delayed up to 1 week after surgery in three of four eyes. The combined use of PTFE and 5-FU or addition of Viscoat allowed adjustment to be delayed for up to 4 weeks after surgery in four of five eyes. CONCLUSION: The combined use of PTFE, 5 FU and Viscoat is useful for delaying suture adjustment in rabbit eyes. PMID- 10845419 TI - Retinopathy of prematurity in pentuplets: outcome and risk factors. PMID- 10845420 TI - Autosomal-dominant megalocornea associated with ocular hypertension. PMID- 10845421 TI - Use of steroids after strabismus surgery. PMID- 10845422 TI - Alstrom syndrome with subclinical insulin-resistant diabetes and hepatic dysfunction: a family report. PMID- 10845423 TI - Accelerated development of polyoma tumors and embryonic lethality: different effects of p53 loss on related mouse backgrounds. AB - Molecular evidence linking polyoma virus to p53 inactivation is thus far lacking, setting this highly oncogenic virus apart from other DNA tumor viruses. As a biological test for interaction, we studied the effects of p53 loss on development of virus-induced tumors. The absence of p53 led to more rapid tumor development on two different mouse backgrounds, indicating synergism between p53 loss and oncogenic pathways controlled directly by the virus. No effects of p53 on tumor type or frequency were noted. Polyoma tumor-derived cells in culture retained p53, and most of these showed induction of p21CIP1/WAF1 in response to DNA damage. These results indicate that p53 functions are not directly and fully impaired by the virus in the intact host. On one mouse background, it was discovered that loss of p53 resulted in complete embryonic lethality prior to 11 days of gestation. This lethality could be rescued by inclusion of gene(s) from a 129/SvJ background. PMID- 10845424 TI - Role of p53 in cell cycle regulation and apoptosis following exposure to proteasome inhibitors. AB - In this study, we explored what effect inhibitors of the 26S proteasome have on cell cycle distribution and induction of apoptosis in human skin fibroblasts and colon cancer cells differing in their p53 status. We found that proteasome inhibition resulted in nuclear accumulation of p53. This was surprising because it is thought that the degradation of p53 is mediated by cytoplasmic 26S proteasomes. Nuclear accumulation of p53 was accompanied by the induction of both p21WAF1 mRNA and protein as well as a decrease in cells entering S phase. Interestingly, cells with compromised p53 function showed a marked increase in the proportion of cells in the G2-M phase of the cell cycle and an attenuated induction of apoptosis after proteasome inhibition. Taken together, our results suggest that proteasome inhibition results in nuclear accumulation of p53 and a p53-stimulated induction of both G1 arrest and apoptosis. PMID- 10845425 TI - Calpain inhibitor 1 activates p53-dependent apoptosis in tumor cell lines. AB - Reports suggest a role of calpains in degradation of wild-type p53, which may regulate p53 induction of apoptosis. A calpain inhibitor, n-acetyl-leu-leu norleucinal (calpain inhibitor 1), was assessed for ability to enhance p53 dependent apoptosis in human tumor cell lines with endogenous wild-type p53 and in altered p53 cell lines with the replacement of wild-type p53 by a recombinant adenovirus (rAd-p53). Calpain inhibitor 1 treatment resulted in increased levels of activated p53, increased p21 protein, and activation of caspases. Cell lines with wild-type, but not mutated or null, p53 status arrested in G0/G1 and were sensitive to calpain inhibitor-induced apoptosis. Regardless of endogenous p53 status, calpain inhibitor treatment combined with rAd-p53, but not empty vector virus, enhanced apoptosis in tumor cell lines. These results demonstrate p53 dependent apoptosis induced by a calpain inhibitor and further suggest a role for calpains in the regulation of p53 activity and induction of apoptotic pathways. PMID- 10845426 TI - Genistein regulation of transforming growth factor-alpha, epidermal growth factor (EGF), and EGF receptor expression in the rat uterus and vagina. AB - Epidemiological reports and laboratory data have associated soy and genistein with reduced incidence of uterine, breast, and prostate cancers, cardiovascular disease and osteoporosis, and lower total blood cholesterol. The aim of this study was to investigate the effect of genistein in the uterus and vagina of rats, focusing our attention on the distribution of transforming growth factor (TGF) alpha, epidermal growth factor (EGF), and EGF receptor. A pharmacological dose of genistein (500 microg/g body weight) injected in rats on days 16,18, and 20 postpartum resulted in significant uterine wet weight gain, with hypertrophy of the luminal and glandular epithelium of the uteri, and squamous epithelium of the vagina in 21-day-old animals. At 50 days of age, hypertrophy was no longer evident in the uterus and vagina. Prepubertal genistein treatment resulted in significantly increased EGF immunostaining in individual stromal cells and reduced EGF receptor immunostaining in blood vessels of the uterus. Genistein treated rats had decreased TGF-alpha immunostaining in glandular and luminal epithelium and a slight increase in EGF receptor immunostaining in stromal cells of the uterus. This suggests paracrine interaction between cells elevating the level of EGF ligand in the stroma and the EGF receptor in the luminal and glandular epithelium, resulting in uterine hypertrophy. In the vagina, genistein did not cause significant alterations to the EGF-signaling pathway in 21- and 50 day-old rats. We conclude that pharmacological doses of genistein during the prepubertal period can modulate the EGF-signaling pathway in the uterus and exert a uterotrophic response in a short-term manner. PMID- 10845427 TI - The novel triterpenoid 2-cyano-3,12-dioxoolean-1,9-dien-28-oic acid induces apoptosis of human myeloid leukemia cells by a caspase-8-dependent mechanism. AB - The oleanane triterpenoid 2-cyano-3,12-dioxoolean-1,9-dien-28-oic acid (CDDO) is a multifunctional molecule that induces growth inhibition and differentiation of human myeloid leukemia cells. The present studies demonstrate that CDDO treatment results in apoptosis of U-937 and HL-60 myeloid leukemia cells. Similar to 1-beta D-arabinofuranosylcytosine (ara-C), another agent that inhibits growth and induces apoptosis of these cells, CDDO induced the release of mitochondrial cytochrome c and activation of caspase-3. Overexpression of Bcl-X(L) blocked cytochrome c release, caspase-3 activation, and apoptosis in ara-C-treated cells. By contrast, CDDO-induced release of cytochrome c, and activation of caspase-3 were diminished only in part by Bcl-X(L). In concert with these findings, we demonstrate that CDDO, but not ara-C, activates caspase-8 and thereby caspase-3 by a cytochrome c-independent mechanism. The results also show that CDDO-induced cytochrome c release is mediated by caspase-8-dependent cleavage of Bid. These findings demonstrate that CDDO induces apoptosis of myeloid leukemia cells and that this novel agent activates an apoptotic signaling cascade distinct from that induced by the cytotoxic agent ara-C. PMID- 10845428 TI - Stem cell factor inhibits erythroid differentiation by modulating the activity of G1-cyclin-dependent kinase complexes: a role for p27 in erythroid differentiation coupled G1 arrest. AB - Terminal erythroid differentiation is accompanied by decreased expression of c Kit and decreased proliferation of erythroid progenitor cells. Using a newly established erythroleukemia cell line HB60-5, which proliferates in response to erythropoietin (Epo) and stem cell factor (SCF) and differentiates when stimulated with Epo alone, we characterized several events associated with the cell cycle during erythroid differentiation. Forty-eight h after SCF withdrawal and Epo stimulation, there was strong inhibition of cyclin-dependent kinase (cdk) 4 and cdk6 activities, associated with an increase in the binding of p27 and p15 to cdk6. A significant increase in the binding of p27 to cyclin E- and cyclin A associated cdk2 correlated with the inhibition of these kinases. In addition, the expression of c-Myc and its downstream transcriptional target Cdc25A were found to be down-regulated during Epo-induced terminal differentiation of HB60-5 cells. The loss of Cdc25A was associated with an increase in the phosphotyrosylation of cyclin E-associated cdk2, which may contribute to cell cycle arrest during differentiation. Although overexpression of p27 in HB60-5 cells caused G1 arrest, it did not promote terminal erythroid differentiation. Thus, the cell cycle arrest that involves p27 is part of a broader molecular program during HB60-5 erythroid differentiation. Moreover, we suggest that SCF stimulation of erythroblasts, in addition to inhibiting erythroid differentiation, activates parallel or sequential signals responsible for maintaining cyclin/cdk activity. PMID- 10845429 TI - Nordic consensus report on asthma management. Nordic Asthma Consensus Group. AB - The work with the Nordic consensus report on asthma management started some years ago. The Nordic countries have common socioeconomic conditions. We acknowledge the international as well as other European guidelines providing valuable recommendations. Nevertheless, we felt the need to combine the common Nordic experiences in order to have a local statement of asthma and asthma care, based upon Nordic clinical science and tradition. The work has been rewarding and we acknowledge many valuable contributions from paediatricians, allergologists and lung physicians in all Nordic countries. The response has so far been positive and we feel that the present material reflects the main opinion of Nordic physicians taking care of asthma patients of all ages. However, the asthma and allergy research field is rapidly developing. Thus, this document should merely be regarded as a time-limited contribution to the continuing scientific discussion of this fascinating field. PMID- 10845430 TI - Control of airway smooth muscle tone. I--electrophysiology and contractile mediators. PMID- 10845431 TI - A comparison of the safety and efficacy of moxifloxacin (BAY 12-8039) and cefuroxime axetil in the treatment of acute bacterial sinusitis in adults. The Sinusitis Study Group. AB - The aim of this multicentre, randomized study was to compare the efficacy and safety of moxifloxacin (BAY 12-8039), a new 8-methoxy fluoroquinolone, with that of cefuroxime axetil for the treatment of acute bacterial sinusitis in adults. Diagnosis was made on a range of clinical signs and symptoms combined with radiology and microbiology. A 400 mg dose of moxifloxacin was administered once daily for 7 days to 242 patients and 250 mg twice daily of cefuroxime axetil was administered to 251 patients for 10 days. The clinical success rate at the end of treatment in the evaluable population was significantly higher (96.7%) in the moxifloxacin group (204/211) than in the cefuroxime axetil group (204/225, 90.7%; 95% confidence intervals 1.5%; 10.6%). At follow-up the success rate in the moxifloxacin group was 90.7% and that for the cefuroxime axetil group was 89.2% (95% confidence intervals -4.3%; 5.4%). The predominant pathogens isolated were Streptococcus pneumoniae and Haemophilus influenzae, followed by Moraxella catarrhalis and Staphylococcus aureus. The bacteriological eradication rates were higher for moxifloxacin (94.5%, 103/109) than for cefuroxime axetil (83.5%, 96/115; 95% CI 3.6%; 19.7%). Only one S. pneumoniae infection persisted following moxifloxacin therapy in contrast with three in individuals on cefuroxime axetil. There were slightly more adverse events in the moxifloxacin group than in the cefuroxime axetil group, but there were fewer serious adverse events following moxifloxacin treatment (three vs. eight). The drug was discontinued because of adverse events in 14 moxifloxacin patients and in 11 cefuroxime axetil patients. Overall, in all assessments, moxifloxacin was at least as effective clinically and bacteriologically, and as well tolerated, as cefuroxime axetil in the treatment of acute sinusitis. PMID- 10845432 TI - Evaluation of CD30 as a marker for th2 lymphocytes in bronchoalveolar lavage in interstitial lung diseases. AB - Several studies have been carried out to clarify the relationship between CD30 expression and Th2 lymphocytes, although the results have been controversial. To investigate whether CD30 is a useful marker for Th2 lymphocytes in bronchoalveolar lavage (BAL) in interstitial lung diseases (ILD), we studied six control subjects and 31 patients with ILD (12 with idiopathic pulmonary fibrosis, seven with hypersensitivity pneumonitis, three with chronic eosinophilic pneumonia and nine with sarcoidosis). The levels of interleukin-5 (IL-5) (secreted by Th2 cells), interferon-gamma (IFNgamma) (secreted by Th1 cells) and the expression of CD30 on lymphocytes were determined in BAL fluid. There were no differences in the percentage of CD30+ lymphocytes between controls and patients with ILD (0.8+0.4% vs. 2+/-0.4%). In order to determine the relationship between Th2 cells and CD30 expression, we divided the patients into two groups according to BAL IL-5 levels. Group I consisted of eight patients (three chronic eosinophilic pneumonia, three hypersensitivity pneumonitis, two idiopathic pulmonary fibrosis) with high IL-5 levels (298+/-138 pg ml(-1)). Group II consisted of the remaining 23 ILD patients with normal IL-5 levels (0.9+/-0.6 pg ml(-1)). The percentage of eosinophils in BAL fluid was significantly higher in group I compared with group 11 (34+/-16% vs. 3+/-1%, P < 0.05). A correlation between CD30+ lymphocytes and IL-5 in group 1 was not shown. There were no differences in the number of CD30+ I we found a significant correlation between IL-5 levels and the percentage of eosinophils (r = 0.95, P < 0.0001). Our results suggest that CD30 does not appear to be a useful marker for Th2 lymphocytes in BAL from patients with ILD. PMID- 10845433 TI - Use of transcutaneous oxygen and carbon dioxide tensions for assessing indices of gas exchange during exercise testing. AB - The slow response characteristics of the combined transcutaneous electrode have been viewed as a major disadvantage when compared with other types of non invasive assessment of gas exchange during exercise testing. We have previously shown that by using the highest recommended temperature of 45 degrees C to reduce response times, and combining this with an exercise protocol of gradual work load increments, that this allows changes in arterial blood gases to be closely followed by transcutaneous values. In the present study we have validated the use of a transcutaneous electrode for estimation of alveolar-arterial oxygen gradient (AaO2) and dead space to tidal volume ratio (V(D)/V(T)) during exercise, against values calculated from direct arterial blood gas analysis. One hundred measurements were made in 20 patients with various cardiopulmonary disorders who underwent exercise testing. Exercise testing was performed by bicycle ergometry with a specific protocol involving gradual work load increments at 2 min intervals. Transcutaneous gas tensions were measured by a heated combined O2 and CO2 electrode. Arterial blood was sampled at the midpoint of each stage of exercise and transcutaneous tensions noted at the end of each stage. The mean difference of the AaO2 gradient calculated from blood gas tensions obtained by the two methods was 0.14 kPa. The limits of agreement were -0.26 and 0.63 kPa. The same values for V(D)/V(T) calculated from gas tensions measured by the two methods were: mean difference 0001; limits of agreement -0.0242 and 0.0252. For both these parameters there was an even scatter around the mean value on Bland and Altman analysis. The findings of this study suggest that estimation of parameters of gas exchange using transcutaneous values during exercise testing is reliable, provided the electrode is heated to a slightly higher temperature than usual and the work load increments are gradual, allowing for the latency in the response time of the system. This system allows the assessment of the contribution of ventilation/perfusion inequality to breathlessness on exertion in patients, provided an initial arterial or ear lobe capillary sample is obtained for calibration purposes. This technique is particularly valuable in patients undergoing repeat exercise tests as it circumvents the need for arterial cannulation. PMID- 10845434 TI - Airway obstruction in relation to symptoms in chronic respiratory disease--a nationally representative population study. AB - We examined the severity of airway obstruction and the occurrence of respiratory symptoms in a large, nationally representative population sample and in a subgroup of subjects with chronic bronchitis and/or emphysema to obtain information for developing national prevention and treatment strategies for these diseases. The study population comprised of 7217 randomly selected subjects (aged 30 years and older) who participated in a comprehensive health examination survey. The 'cases' were subjects diagnosed as having chronic bronchitis and/or emphysema. The survey methods comprised of questionnaires, interviews, physical measurements, including spirometry, and clinical examinations. In the whole study population, the age-adjusted prevalence of chronic bronchitis and/or emphysema was 22% among men and 7% among women, whilst clinically relevant airways obstruction (FEV1/FVC%< or = 69) was present in 11% of men and in 5% of women. The occurrence of chronic cough and phlegm production was lowest among the 'cases' with pronounced obstruction (in 68% of men with severe and in 60% of women with moderate obstruction), whereas cold air-associated dyspnoea aggravation showed an inverse relationship, occurring most commonly in men (80%) with severe obstruction. Unexpectedly, half of the bronchitic women had never smoked. We conclude that the occurrence of certain bronchitic symptoms, such as chronic cough and phlegm production and cold air-associated dyspnoea aggravation, may to some degree indicate different stages of the disease. Smoking was not closely associated with airflow limitation in women here. PMID- 10845435 TI - In vivo efficacy of heated and non-heated humidifiers during nasal continuous positive airway pressure (nCPAP)-therapy for obstructive sleep apnoea. AB - Upper airway dryness is a frequent side-effect of nasal continuous positive airway pressure therapy (nCPAP) in obstructive sleep apnoea (OSA). In this situation, heated or non-heated passover humidifiers are often added to the nCPAP therapy. The efficacy of these two modes in terms of increasing the absolute humidity of the inspired air in vivo has so far not been established. The present investigation was therefore designed to compare various heated and non-heated passover humidifiers in terms of the their ability to increase the absolute humidity in the inspired air during nCPAP. In six healthy test individuals, nCPAP therapy at pressures of 5 mbar and 10 mbar was simulated, and the relative humidity and temperature of the air within the tube at the junction between CPAP tube and mask were measured. In each test person, measurements were carried out both with and without the two heated (HC 100, Fischer&Paykel Inc., New Zealand and HumidAire, ResMed Ltd., Australia) and two non-heated (Oasis and Humidifier, both from Respironics Inc., U.S.A.) passover humidifiers under steady-state conditions. The absolute humidity was calculated from the relative humidity and temperature measurements. The mean (SD) absolute humidity (gm(-3)) in the steady state was significantly (P<0.05 higher with each of the humidifiers than that calculated when no humidifier was used. The relevant figures were as follows: no humidifier: 10(-2) (1.8) gm(-3) (at 5 mbar)/9.8 (1.8) gm(-3) (at 10 mbar); Humidifier: 16.4 (0.97)/15.6 (1.26); Oasis: 17.3 (0.97)/ 16.7 (0.93); HC100: 26.5 (1.40)/26.2 (1.23); HumidAire: 31.8 (2.50)/30.9 (2.64). The mean increase in absolute humidity (in gm(-3)) with the aid of the heated humidifiers was 16.3 (5 mbar) gm(-3)/16.4 (10 mbar) gm(-3) with HC100 and 21.6/21.1 with HumidAire, and in both cases was clearly and significantly (P=0.028) higher in comparison with the non-heated humidifiers--6.2/5.8 with Humidifier and 7.2/6.9 with Oasis. In terms of the absolute humidity achieved within the CPAP tube system, the heated humidifiers were clearly superior to the non-heated humidifiers. These results were, however, obtained under laboratory conditions, and therefore cannot be translated unreservedly to the situation represented by long-term CPAP-treatment. Furthermore, it is possible that the smaller humidification capacity of the non heated humidifiers may still suffice to meet the requirements of clinical use in terms of effectively preventing dry airways under CPAP treatment. This point, however, needs further investigation on the basis of long-term clinical studies. PMID- 10845436 TI - Treatment of endobronchial metastases with intraluminal radiotherapy. AB - Metastasis to the lung occurs quite commonly from certain types of extrapulmonary primary carcinoma. Spread to the bronchial lumen is relatively rare. When this does occur, symptoms resembling those of primary bronchial carcinoma are often present, in association with partial or complete obstruction of the bronchial lumen. Palliation of such symptoms is possible with the use of intraluminal radiotherapy (ILT). Between 1990 and 1998, 37 patients with endobronchial metastases were treated using this modality; a single fraction of radiation was delivered by the remote afterloading high dose rate microSelectron system. Data regarding these patients' characteristics and outcome are presented, following a retrospective review of case notes. The commonest symptoms were dyspnoea, cough and haemoptysis; the commonest primary tumour sites were breast, colorectum, oesophagus and kidney. Twenty-four (64.9%) patients had some improvement in symptoms following treatment. Mean overall survival was 280 days, range 9-1145 days. No serious adverse effects occurred. ILT is a relatively simple, safe and effective treatment in the palliation of symptoms due to endobronchial metastases. PMID- 10845437 TI - Accuracy of recorded asthma deaths in Denmark in a 12-months period in 1994/95. AB - Many studies of asthma mortality rely on official registration. The aim of this study was to evaluate the accuracy of death certificates, where asthma was coded as cause of death. In a 12-month period, medical information on all subjects with asthma officially coded as the underlying cause of death in Denmark, was obtained by reviewing hospital records, contacting general practitioners and sometimes close relatives. A panel of four pulmonologists each examined the obtained information and independently assessed the cause of death. Of a total of 218 death certificates, 39 were excluded as the cause of death could not be validated. In 16 (9%) of the subjects death from asthma was judged to be the definite cause of death and in 12 (7%) death from asthma was possible. Of 151 non asthma deaths coded as due to asthma, 109 were judged to have suffered or died from COPD and 14 from heart disease. The accuracy of Danish death certification in asthma deaths is poor, especially in the elderly, where COPD is often classified as asthma. We conclude that the true asthma mortality in Denmark is substantially lower than officially recorded. PMID- 10845438 TI - Effect of early ambroxol treatment on lung functions in mechanically ventilated preterm newborns who subsequently developed a bronchopulmonary dysplasia (BPD). AB - In a randomized trial in 102 preterm newborns with respiratory distress syndrome (RDS) it has been shown that early Ambroxol treatment (30 mg kg(-1) over the first 5 days) significantly reduces the incidence of RDS-associated complications [bronchopulmonary dysplasia (BPD), intraventricular haemorrhage, post-natal acquired pneumonia]. The aim of the present analysis was to investigate the effect of Ambroxol treatment on lung function in newborns who developed BPD. Respiratory function testing (RFT) was performed immediately after extubation and at day 28. Tidal volume (VT) and respiratory frequency (f) were measured during tidal breathing using the deadspace free flow-through technique. The lung mechanic parameter VT/maxPes was determined by measuring the maximal oesophageal pressure changes, maxPes, with a catheter tip pressure transducer. In the placebo group 36/50 infants were extubated within the first 28 days of life and 13/36 (36%) developed BPD. In the Ambroxol group 44/52 were extubated and 9/44 (20%) developed BPD. After extubation, RFT showed (i) no statistically significant difference in the ventilatory parameters of either treatment group, (ii) improved (P<0.05) lung mechanics (VT/maxPes) in Ambroxol group compared to controls (94+/ 27 ml kPa(-1) vs. 8.1+/-2.6 ml kPa(-1)) and (iii) no statistically significant difference in lung function between infants with and without BPD. At day 28 we found (i) no effect of early Ambroxol treatment on lung functions, (ii) significantly (P < 0.05) higher f (58.5+/-11.7 min(-1) vs. 49.7+/-10.1 min(-1)) and significantly (P<0.01) lower V(T) (9.6+/-1.9 ml vs. 12.3+/-2.7 ml) and V(T)/maxPes (8.9+/-2.6 ml kPa(-1)] vs. 12.0+/-2.9 ml kPa(-1)) in infants with BPD compared to infants without and (iii) these differences are not influenced by early Ambroxol treatment. If the process of BPD development is induced, early Ambroxol treatment has no influence on impaired lung function at day 28. PMID- 10845439 TI - Timing of lowest and highest peak expiratory flow in patients with asthma: influence of anti-inflammatory treatment. AB - We sought to determine the optimal time for measuring peak expiratory flow rate (PEF) in patients with mild to moderate asthma, before and after treatment with inhaled beclomethasone dipropionate (BDP). After 2 weeks of observation, BDP (400 microg/d) was given to 22 patients with mild to moderate asthma. The dose of BDP (800-1200 microg/d) was increased every 2 weeks until PEF varied by no more than 20% each day. PEF was measured four times daily: on awakening, around noon, in the evening and at bedtime. Significant (P < 0.05) rhythms were detected by single cosinor analysis in all patients, both during observation and during treatment. Analysis by the population mean-cosinor method showed that the mean mesor was 378.8+/-59.1 lmin(-1), the mean amplitude was 53.9+/-13.4 lmin(-1), and the mean acrophase was at 16:26+/-0:32 before treatment. After treatment, the mean mesor was 528.0+/-61.9 l min(-1), the mean amplitude was 37.6+/-12.2 lmin( 1), and the mean acrophase was at 16:35+/-0:32. The mesor increased significantly (P<0.05), and the amplitude decreased significantly (P<0.05) after treatment. The acrophase did not change. These data indicate that PEF is lowest at 04:30 and highest at 16:30 in patients with mild to moderate asthma, both during observation and during treatment. We conclude that if one needs to assess PEF twice a day, this should ideally be done at 04:30 and 16:30, not only before but also after treatment with BDP. PMID- 10845440 TI - Smoking habits in a cohort of U.K. adolescents. AB - Active smoking is an increasing problem amongst U.K. teenagers. The smoking habits of a cohort of 14-16-year-olds were determined and the association between regular active smoking and domestic and social factors investigated. Current smoking habits of a cohort of 2289 14-16-year-olds were assessed using a simple postal questionnaire. Data concerning potential factors associated with active smoking were collected from questionnaire completed by parents. Nine hundred and sixty-nine (44.8%) children admitted to having smoked at some time, with 562 (30.0%) having smoked in the previous 12 months. Three hundred and six (14.1%) children were regular smokers and 158 (51.6% of regular smokers, 7.3% of total cohort) smoked daily. Age, number of other children in the household, parental smoking, smoking sibling(s) and living in a single parent household were all independently associated with regular smoking. Regular smoking was a significant problem amongst this cohort of teenagers. Living with other smokers, age, household size and living with one parent all predicted a regular smoking habit. PMID- 10845441 TI - Repeatability of the ISAAC video questionnaire and its accuracy against a clinical diagnosis of asthma. AB - The objective of the study was to evaluate the performance of the International Study of Asthma and Allergies in Childhood (ISAAC) video questionnaire in terms of repeatability and accuracy against a clinical diagnosis of asthma achieved according to the National Heart, Lung and Blood Institute (NHLBI) algorithm. Two hundred and forty-one subjects, aged 13-14 years from two secondary schools in Rome, Italy, were enrolled. Video and written ISAAC questionnaires were completed twice, 3 months apart, by 194 and 190 adolescents, respectively. Two months later, 106 subjects were visited by two physicians blinded to the results of questionnaires. Sixteen subjects were classified as having clinical asthma (CA) at the clinical visit, and eight of them as having clinical active asthma (CAA) on the basis of at least one positive outcome of the NHLBI algorithm. The repeatability of video questionnaire was similar to that of the written questionnaire for items on exercise wheeze and nocturnal cough and, to a lesser degree, for items concerning any wheeze in the past. The video questionnaire showed a worse performance than the written questionnaire for items on asthma attack: K-value (95% CL) = 0.59 (0.37-0.80) for video scene no. 5 and K-value (95% CL) = 0.86 (0.74-0.98) for written question no. 6. The overall accuracy of the video questionnaire, estimated as a positive answer to any video scene, was lower in terms of sensitivity than that of any written question when CA was used as a gold standard (0.50 vs. 0.81, P=0.025) and increased with respect to CAA (0.75 vs. 0.87, P = 0.317). The specificity of any video scene was better than that of any written question, independently from the gold standard used. In conclusion, the video questionnaire showed a fairly good accuracy, although slightly lower than that of the written questionnaire and provided sufficiently reliable results. However, samples of subjects from different geographic areas and cultures should be studied in order to conclusively define the performance of the ISAAC video questionnaire. PMID- 10845442 TI - Untreated idiopathic aneurysm of the pulmonary artery; long-term follow-up. AB - A 57-year-old man with an idiopathic aneurysm of the left pulmonary artery is presented. The aneurysm was stable over many decades. No such long-term follow-up of an idiopathic pulmonary aneurysm has been described previously. After 39 years there is no evidence of heart failure or pulmonary hypertension. Because corrective surgery has a variable outcome, and prognosis is suggested to be good in the idiopathic form of pulmonary aneurysm, no surgical correction was proposed. PMID- 10845443 TI - Association of 210Po(210Pb), 239+240Pu and 241Am with different mineral fractions of a beach sand at Seascale, Cumbria, UK AB - The degree of association of 210Po (in secular equilibrium with 210Pb), 239+240Pu and 241Am with iron oxide minerals in beach sand has been assessed using samples collected by panning and selective magnetic extraction techniques. The samples were obtained from Seascale beach (Cumbria), close to the BNF reprocessing plant at Sellafield (UK) and physically and chemically characterised in terms of their grain-size distribution, radionuclide content and magnetic properties. Low frequency (0.46 kHz) susceptibility (X(LF)) and saturation isothermal remanent magnetisation (SIRM) measurements were used to determine the magnetic properties of the bulk samples and the grain size fractions. Samples were also subjected to magnetic extraction, to concentrate the iron oxide minerals, and thence determine their degree of association with the radionuclides of interest. The efficiency of the magnetic extraction technique was estimated from rock magnetic measurements, before and after the extraction procedure. The placer deposits, enriched in iron oxide grains by environmental sorting processes, were both finer grained and more magnetic than the bulk sand. The mean particle diameter of the bulk sand was largely (approximately 75%) greater than 200 microm, whereas for the placer material, > 88% of particles have mean diameters of < 200 microm. Whilst X(LF) and SIRM values of the 100-200 microm placer material were approximately 40-50 fold higher than those of the bulk sand of equivalent grain size, the radionuclide concentrations were approximately 4-13-fold higher. In terms of mass, the percentage of magnetic material extracted from samples of bulk sand (using a powerful, approximately 0.3 T, rare earth magnet), was small (approx. 1 3%). Nevertheless, > 70% of the magnetic signal was associated with these extracts. Concentrations of all three radionuclides in the magnetic extracts were enhanced relative to the weakly magnetic residue. The proportions of 210Po(210Pb), 239+240Pu and 241Am associated with the magnetic extracts were approximately 18%, approximately 12% and approximately 11%, respectively. Therefore, although the strongly magnetic iron oxide fraction contains measurably enhanced levels of these radionuclides, the majority of the alpha-radioactivity appears to be associated with the weakly magnetic residue. PMID- 10845444 TI - Reconstructing historical radionuclide concentrations along the east coast of Ireland using a compartmental model. AB - A mathematical model is presented that simulates the annually averaged transport of radionuclides, originating from the BNFL reprocessing plant at Sellafield, throughout the Irish Sea. The model, CUMBRIA77, represents the processes of radionuclide transport and dispersion in the marine environment and allows predictions of radionuclide concentration in various environmental media, including biota, to be made throughout the whole of the Irish Sea. In this paper we describe the use of the model to reconstruct the historical activity concentrations of 137Cs and 239+240Pu in a variety of environmental media in the western Irish Sea and along the Irish east coast back to 1950. This reconstruction exercise is of interest because only limited measurements of 137Cs and 239+240Pu activity are available prior to the 1980s. The predictions were compared to the available measured data to validate their accuracy. The results of the reconstruction indicate that activity concentrations of 137Cs in the western Irish Sea follow a similar, though slightly delayed and smoothed, profile to the discharges from the Sellafield site, with concentrations at the time of peak discharge (the mid-1970s) being around an order of magnitude higher than those measured in the 1980s and 1990s. By contrast, the concentrations of 239+240Pu at the time of peak discharges were similar to those presently measured. These differences reflect the distinct marine chemistries of the two nuclides, in particular the higher propensity of plutonium to bind to sediments leading to extended transport times. Despite these differences in behaviour the doses to Irish seafood consumers from 137Cs remain significantly higher than those from 239+240Pu. PMID- 10845445 TI - An extended study of heavy metal deposition in Galicia (NW Spain) based on moss analysis. AB - This paper describes the first attempt to determine the levels of heavy metal contamination in Galicia (NW Spain) by analysis of moss species. Samples of Scleropodium purum and Hypnum cupressiforme were collected from 75 sites, in 1995 and 1997, and analysed by atomic absorption spectrophotometry and atomic fluorescence for Al, As, Co, Cr, Cu, Fe, Hg, Ni, Pb and Zn. Comparisons were made between the two moss species used and, for those elements present at different concentrations in the two species, regression lines were made for interspecies calibration. Distribution maps were drawn up for each of the elements according to their concentration in the moss. In certain areas it was possible to relate deposition to the existence of known sources of contamination. The study demonstrates that the highest levels of elements were found in the most heavily industrialised and populated areas. Concentrations of As, Cu, Hg and Ni, recorded during the 1997 sampling were significantly higher than those found in 1995, possibly due to agricultural and industrial activities. PMID- 10845446 TI - Aluminium and iron burdens of aquatic biota in New Zealand streams contaminated by acid mine drainage: effects of trophic level. AB - Concentrations of Al and Fe were determined in samples of filamentous algae, bryophytes and invertebrates from 24 stream sites in North Westland, South Island, New Zealand. Sites were variably contaminated by acid coal mine drainage and ranged in pH from 2.6 to 6.2. Conductivity of stream water ranged from 16 to 944 microS25 cm(-1) and maximum concentrations of total dissolved Al and total Fe measured in two successive years were 35.5 and 32.6 mg l(-1), respectively. Metal burdens of algae and bryophytes were not correlated with pH, conductivity or the concentrations of Al and Fe observed in stream water. Metal concentrations in invertebrates were significantly lower than those in plants (mg per g dry wt.), and were similar in herbivore-detritivores (mainly mayfly larvae) and carnivorous species. No evidence was found for the biomagnification of either metal within aquatic food webs. However, invertebrate species exposed to very high concentrations of Al and Fe varied considerably in body burdens, suggesting that groups of insects differ considerably in their physiological or morphological ability to exclude potentially toxic metals. PMID- 10845447 TI - Calculation and mapping of critical loads of sulfur and nitrogen in Flanders, Belgium. AB - Up to now, critical loads calculations for the Flemish Region were based on European background data of surrounding countries. A first attempt has been made to calculate and map critical loads for forest ecosystems in Flanders using available site-specific information. Values of current deposition were used to calculate and map exceedances. The lowest critical loads for acidification (697 eq ha(-1)year(-1)) occur in the Campine and the north of Limburg where ecosystems largely consist of coniferous forests on poor sandy soils. The dominance of coniferous forest types in the Campine is also responsible for low critical load values for eutrophication (between 536 and 971 eq ha(-1)year(-1)). In 75% of the receptor points that have been considered an exceedance of the critical load for acidification is noted, primarily in areas with high SO2 and NOx depositions, such as the north of the provinces East and West Flanders and Antwerp. The critical load for eutrophication is exceeded in all points considered. Exceedances are particularly high in coniferous forests in West Flanders, and in the north of the provinces of Antwerp and Limburg, where especially NHx depositions amount to high values. Data needed for the calculation of critical loads are still sparse in Flanders, e.g. for: (1) weathering rates of soil minerals; (2) interception and evaporation of forest ecosystems; and (3) uptake of N and basic cations by vegetation. This supplementary information will contribute to a further refining of the calculated critical loads, which constitute indispensable information in developing an emission abatement policy. PMID- 10845448 TI - Measurements of N2O and CH4 from the aerated composting of food waste. AB - Emissions of N2O and CH4 from an aerated composting system were investigated using small-scale simulated reactors. The results show relatively high emissions of N2O at the beginning of composting, in proportion to the application amount of food waste. After 2 days, the N2O emission decreased to 0.53 ppmv on average, near to the background level in the atmosphere (0.45 ppmv). The addition of composted cattle manure increased N2O emissions not only at the beginning of composting, but also during the later period and resulted in two peak emission curves. Good correlation was observed between the N2O concentration at the air outlet and NO2- concentration in waste, suggesting a generation pathway for N2O from NO2- to N2O. Methane was only detected in treatments containing composted cattle manure. The high emission of methane illustrates the involvement of anoxic/anaerobic microorganisms with the addition of composted manure. The result suggests the existence of anoxic or anaerobic microsite inside the waste particles even though ventilation was employed during the composting process. PMID- 10845449 TI - Solid phase partitioning of metals in managed retreat soils: field changes over the first year of tidal inundation. AB - Sequential extraction methods were used to determine the effect of saline inundation on partitioning behaviour of metals in soils at Orplands Managed Retreat site, Essex, UK. A suite of metals (Al, Ca, Cr, Cu, Fe, K, Li, Mn, Ni, Pb and Zn) were analysed in sequential extracts from samples of soil cores extracted immediately before breaching of sea walls in 1995 and 1 year after. Generally, partitioning of most metals was dominated by associations with the residual fraction and an absence of associations with the exchangeable fraction indicating a relatively uncontaminated environment. Changes in partitioning were predominantly limited to the top 8 cm of the soil profile where a decaying mat of vegetation from the pre-inundation surface resulted in high moisture contents and low redox potentials. In this zone, the predominance of metal associations with the residual fraction was replaced by associations with more labile fractions. In the surface layer deposited after breaching, Ca was dominantly associated with the carbonate fraction, which indicates biogenic import from adjacent estuarine sediments. PMID- 10845450 TI - Role of organic acids in detoxification of aluminum in higher plants. AB - Phytotoxicity of aluminum ion (Al3+) is a serious problem limiting crop production on acid soils. Organic acids with Al-chelating ability play an important role in the detoxification of Al both externally and internally. Al is detoxified externally by the secretion of organic acids such as citric, oxalic, and/or malic acids from the roots. The secretion of organic acids is highly specific to Al and the site of secretion is localized to the root apex. The kind of organic acids secreted as well as secretion pattern differ among plant species. There are two patterns of Al-induced secretion of organic acids: In pattern I, there is no discernible delay between the addition of Al and the onset of the release of organic acids. Activation of the anion channel seems to be involved in this pattern; In pattern II, there is a marked lag phase between the addition of Al and the onset of organic acid release. The action of genes related to the metabolism and secretion of organic acids seems to be involved in this pattern. Internal detoxification of Al in Al-accumulating plants is achieved by the formation of Al-organic acid complex. For instance, the complex of Al-citrate (1:1) in hydrangea and Al-oxalate (1:3) in buckwheat has been identified. PMID- 10845451 TI - Involvement of jasmonate- and salicylate-related signaling pathways for the production of specific herbivore-induced volatiles in plants. AB - We compared volatiles from lima bean leaves (Phaseolus lunatus) infested by either beet armyworm (Spodoptera exigua), common armyworm [Mythimna (Pseudaletia) separata], or two-spotted spider mite (Tetranychus urticae). We also analyzed volatiles from the leaves treated with jasmonic acid (JA) and/or methyl salicylate (MeSA). The volatiles induced by aqueous JA treatment were qualitatively and quantitatively similar to those induced by S. exigua or M. separata damage. Furthermore, both S. exigua and aqueous JA treatment induced the expression of the same basic PR genes. In contrast, gaseous MeSA treatment, and aqueous JA treatment followed by gaseous MeSA treatment, induced volatiles that was qualitatively and quantitatively more similar to the T. urticae-induced volatiles than those induced by aqueous JA treatment. In addition, T. urticae damage resulted in the expression of the acidic and basic PR genes that were induced by gaseous MeSA treatment and by aqueous JA treatment, respectively. Based on these data, we suggest that in lima bean leaves, the JA-related signaling pathway is involved in the production of caterpillar-induced volatiles, while both the SA-related signaling pathway and the JA-related signaling pathway are involved in the production of T. urticae-induced volatiles. PMID- 10845452 TI - Possible involvement of phosphoinositide-Ca2+ signaling in the regulation of alpha-amylase expression and germination of rice seed (Oryza sativa L.). AB - We have studied the effects of neomycin, a potent inhibitor of inositol phospholipid-specific phospholipase C (PLC), on the germination of rice seed and the gibberellin-induced expression of alpha-amylase in the aleurone layer and the scutellar tissues. It was shown that, in the absence of exogenous Ca2+, neomycin markedly reduced the germination speed and seedling growth of rice seeds and inhibited the gibberellin-induced expression of alpha-amylase in both secretory tissues. In addition, neomycin decreased the formation of inositol 1,4,5 trisphosphate (IP3) in the gibberellin-treated aleurone layer and the scutellar tissues. However, the inhibitory effects on the germination speed and the expression of alpha-amylase activity were overcome by supplementation of Ca2+. In addition, gibberellin elevated the level of IP3, and ABA prevented the gibberellin-induced formation of IP3, although ABA alone did not alter the IP3 level. The expression of a membrane-bound PLC molecule in rice aleurone layer was shown to be induced by gibberellin, and the gibberellin-induced expression of PLC was markedly delayed by treatment with ABA. These results strongly suggest that the phosphoinositide-Ca2+ signal transduction pathway may play an important role in the gibberellin-induced expression of alpha-amylase molecules closely related to the germination processes of rice seed. PMID- 10845453 TI - Purification and characterization of two sucrose synthase isoforms from Japanese pear fruit. AB - Two isoforms (SS I and SS II) of sucrose synthase (SS; EC 2.54.1.13) were purified from Japanese pear fruit and their properties were compared. SS I mainly appeared in young fruit and SS II mainly in mature fruit. SS I and SS II were purified to the specific activity of 3.37 and 4.26 (units (mg protein)(-1)), respectively. The MW of native and subunit proteins of SS I and SS II were almost the same and both SSs seemed to be a tetramer composed of an 83 kDa polypeptide. However, the ionic charges of the native proteins and the kinetic parameters of SSs were different. Specifically, the Km value for UDP-glucose in SS I was the same as that for UDP, while the Km value for UDP-glucose in SS II was less than that for UDP. SS II easily reacted for sucrose synthesis than sucrose cleavage compared with SS I. Therefore, it is considered that SS I and SS II play different roles in the utilization of carbohydrate in young and mature fruit, respectively. PMID- 10845454 TI - Rice NPH1 homologues, OsNPH1a and OsNPH1b, are differently photoregulated. AB - The members of nonphototropic hypocotyl 1 (NPH1) family of genes in plants are considered to be the blue-light photoreceptors for phototropism. We isolated and characterized two NPH1 homologues from rice named as OsNPH1a and OsNPH1b. The predicted proteins of both OsNPH1 genes include LOV (LOV1 and LOV2) and protein kinase domains which are typically conserved regions in all the members of NPH1 family. Comparison of OsNPH1 apoproteins with those from other plant species revealed a close homology of OsNPHla with Arabidopsis, maize and oat NPH1, while the OsNPH1b appeared to be more closer to the recently found Arabidopsis NPL1. These structural homologies indicate that NPH1 homologues can be grouped into two classes namely "NPH1 type" and "NPL1 type". Northern blot analysis showed that OsNPH1a was strongly expressed in coleoptiles, whereas OsNPH1b was highly expressed in leaves of dark-grown rice seedlings. When the dark-grown seedlings were transferred to the continuous white light, the abundance of the OsNPH1a transcript in coleoptiles rapidly declined to the minimum levels, whereas the OsNPH1b transcript level in leaves gradually increased. These results lead us to conclude that expression of OsNPH1a and OsNPH1b is differently photoregulated in different tissues of rice seedlings. PMID- 10845455 TI - Regulation of cytoplasmic pH under extreme acid conditions in suspension cultured cells of Catharanthus roseus: a possible role of inorganic phosphate. AB - Changes in cytoplasmic pH of suspension-cultured cells of Catharanthus roseus under extreme acid conditions were measured with the pH-dependent fluorescence dye; 2',7'-bis-(2-carboxyethyl)-5 (and-6) carboxyfluorescein ( acetoxymethylester) (BCECF). When cells were treated with 1 mM HCl (pH 3 solution), the cytoplasmic pH first decreased then returned to the original level. Treatment with 10 mM HCl (pH 2 solution) acidified the cytoplasm to a greater extent, and the acidification continued at a constant level throughout the measurement. Treatment with a pH 2 solution resulted in a gradual decrease of the malate content, indicating the operation of biochemical pH regulation mechanism. The pH 2 treatment also caused a sudden decrease of the intracellular level of Pi. The cellular content of total phosphorus did not change during the acidification. The Pi was converted to the organic phosphate form. The ATP level was not increased by the pH 2 treatment, but slightly decreased. The role of Pi, which might be functioning as a regulatory factor of cytoplasmic pH, a non competitive inhibitor of the H+-pumps of both the plasma membrane and tonoplast is discussed. PMID- 10845456 TI - Photosynthetic parameters of leaves of wild type and Cyt b6/f deficient transgenic tobacco studied by CO2 uptake and transmittance at 800 nm. AB - Parallel measurements of CO2 assimilation and 800 nm transmission were carried out on intact leaves of wild type and cytochrome b6/f deficient transgenic tobacco grown at different light intensities and temperatures, with the aim to diagnose rate-limiting processes in photosynthesis and investigate their adaptations to growth conditions. Maximum CO2- and light-saturated photosynthetic rate, mesophyll conductance, assimilatory charge and specific carboxylation efficiency were determined from CO2 fixation measurements and postillumination P700 rereduction time constant was measured from the transient of the 800 nm signal. Results show that growth conditions continue to modulate the expression of genes in transgenic plants, interfering with the antisense modulation, but under all environmental conditions the antisense treatment to decrease Cyt b6/f complexes ensured that the control of electron/proton transport rate by proton backpressure on the PSI donor side was stronger than the control by electron backpressure on the PSI acceptor side. Coordinated control of gene expression and enzyme activation ensures that different parts of the photosynthetic machinery- components of the electron transport chain, ribulose-1,5-bisphosphate carboxylase/oxygenase, enzymes of the sucrose and starch synthesis chains-are synthesized more or less proportionally under different environmental conditions and in case of mild genetic interference. PMID- 10845457 TI - Wound-induced ethylene synthesis and expression and formation of 1 aminocyclopropane-1-carboxylate (ACC) synthase, ACC oxidase, phenylalanine ammonia-lyase, and peroxidase in wounded mesocarp tissue of Cucurbita maxima. AB - 1-Aminocyclopropane-1-carboxylate (ACC) synthase was rapidly induced in mesocarp tissue of Cucurbita maxima after wounding in the cut surface layer in 1 mm thickness (ca. 9 cells) (first layer) in both the enzyme activity and the levels of transcript. This led to a rapid accumulation of ACC and hence ethylene production. In the inside tissue (1-2 mm) (second layer), no significant induction of ACC synthase was observed, which resulted in a low level of ACC, although ethylene was evolved at a much lower rate than the first one. In contrast to ACC synthase, ACC oxidase was induced markedly in both the first and second layers and the development of its activity and the levels of mRNA remained high until later stages. It was considered that wound ethylene was closely associated with the development of ACC oxidase, since 2,5-norbornadiene (NBD), an inhibitor of ethylene action, substantially suppressed it. Phenylalanine ammonia lyase (PAL) greatly increased in activity after wounding similarly to that of ACC synthase, in which increase in PAL activity occurred predominantly in the first layer. Induction of peroxidase activity after wounding had a close correlation in profile with that of ACC oxidase in that marked increases in the activity were observed in both the first and second layers and were strongly suppressed by NBD application. Four peroxidase isozymes were found by PAGE, among which a fraction was newly detected after wounding. PMID- 10845458 TI - A tobacco NtMET1 cDNA encoding a DNA methyltransferase: molecular characterization and abnormal phenotypes of transgenic tobacco plants. AB - A cDNA encoding a DNA methyltransferase, with a predicted polypeptide of 1556 amino acid residues containing all motifs conserved in this enzyme family, was isolated from tobacco plants, and the corresponding gene was designated as NtMET1. RNA blot analysis indicated NtMET1 transcripts to accumulate in dividing tissues of tobacco plants, and they could be detected during the S phase in synchronized dividing BY2 cells. In situ hybridization revealed the transcripts to be localized exclusively in actively proliferating tissues around axillary apical meristem. In order to ascertain physiological roles, transgenic tobacco plants that had the antisense construct were made and examined for phenotypes. Methylation levels of genomic DNA from transgenic plants significantly decreased in comparison with wild-type levels, and distinct phenotypic changes including small leaves, short internodes and abnormal flower morphology were noted. Microscopic observation revealed that leaf structure differed between transgenic and wild-type plants. These results suggest that NtMET1 functions during DNA replication, and that DNA methylation plays an important role in plant morphogenesis. PMID- 10845459 TI - Isolation of the gene for EILP, an elicitor-inducible LRR receptor-like protein, from tobacco by differential display. AB - We screened tobacco genes, which are differentially expressed in response to a fungal elicitor, and have isolated a gene which codes for a leucine-rich repeat (LRR) protein closely related to Cf genes in tomato. The EILP (elicitor inducible LRR protein) gene encodes 95 kDa protein, which consists of a putative membrane spanning region, 28 leucine-rich repeats and some N-linked glycosylation sites, and shows high homology to Cf-2/Cf-5 family genes. Southern blot analysis revealed the presence of some genes homologous to EILP in tobacco, like Cf genes in tomato. The expression of EILP was low at the basal level and increased by treatment with elicitor, implying that EILP is involved in both preexisting and inducible surveillance systems. The expression of EILP was activated by a non pathogen, Pseudomonas syringae pv. glycinea, and in a delayed fashion by the tobacco pathogen P. syringae pv. tabaci, suggesting that the product of EILP may be involved in non-host disease resistance in tobacco. PMID- 10845460 TI - Three Arabidopsis genes encoding proteins with differential activities for cysteine synthase and beta-cyanoalanine synthase. AB - Three cDNA clones encoding putative cysteine synthases (O-acetylserine (thiol) lyase, EC 4.2.99.8) were isolated from Arabidopsis thaliana and designated AtcysC1, AtcysD1 and AtcysD2, respectively. Southern blot analyses suggested that the corresponding genes were present as a single copy, or at most two copies, in the A. thaliana genome. Escherichia coli complementation analyses confirmed that the cDNAs encode cysteine synthase and the corresponding proteins produced in E. coli clearly showed cysteine synthase activity. In addition, AtcysC1 protein showed beta-cyanoalanine synthase (EC 4.4.1.9) activity, but the other two did not. Kinetic analysis suggests that AtcysC1 actually functions as beta cyanoalanine synthase rather than cysteine synthase in vivo. The mRNA accumulation of AtcysC1, AtcysD1 and AtcysD2 differed in various organs, but did not change markedly when A. thaliana seedlings were subjected to various stresses, including nutrient deprivation. In vivo targeting experiments indicated that AtcysD1 and AtcysD2 are cytoplasmic isozymes, and AtcysC1 is a mitochondrial isozyme. PMID- 10845461 TI - A pollen-specific and desiccation-associated transcript in Lilium longiflorum during development and stress. AB - A cDNA clone encoding a desiccation-induced protein (LLA23) has been isolated from the mature pollen of a Lilium longiflorum cDNA library. Sequence analysis revealed significant similarity between the predicted LLA23 polypeptide, particularly at the C-terminal half of the sequence and a group of water deficit/ripening-induced proteins. The expression of LLA23 gene is pollen specific and the transcript accumulates only at the later stage of pollen maturation prior to anthesis. Premature drying of developing pollen confirmed that the accumulation of LLA23 transcripts was associated with desiccation. The LLA23 proteins decreased their levels when pollen/pollen tubes grew in the germination buffer. Treatments of pollen with abscisic acid (ABA) and polyethylene glycol (PEG)-8000 during germination greatly retarded the disappearance of LLA23 proteins and mRNAs. The LLA23 transcripts decreased their levels in pollen tubes grown both in vitro and in vivo, but the disappearance of LLA23 transcripts in tube cells grown in vivo was slower than those grown in vitro. In situ localization using anti-chicken immunoglobulin G conjugated with gold particles confirmed that LLA23 was located in the cytoplasm of pollen grains. The protective function of the desiccation-related proteins in the cytoplasm of pollen grains is proposed. PMID- 10845462 TI - Characterization of cross-links between cellulose microfibrils, and their occurrence during elongation growth in pea epicotyl. AB - The occurrence and chemical nature of the cross-links between cellulose microfibrils in outer epidermal cell walls in Pisum sativum cv. Alaska was investigated by rapid-freezing and deep-etching techniques coupled with chemical and enzymatic treatments. The cell wall in the elongating region of epidermal cells was characterized by the absence of the cross-links, while in the elongated region, the cell wall was characterized by the presence of cross-links. The cross links remained in the cell wall of the elongated region after treatment with SDS electrophoresis sample buffer and treatment with 4% potassium hydroxide. After treatment with endo-1,4-beta-glucanase, which fragments xyloglucan, the cross links were remarkably reduced from the cell wall of the elongated region. The endoglucanase treatment also reduced immunogold labeling of xyloglucan in the cell wall. The endoglucanase hydrolysate from the cell wall fraction of the elongated region gave spots of oligosaccharides in thin layer chromatography, which were identical to the spots of xyloglucan oligosaccharides produced by xyloglucanase from both the cell wall fraction and tamarind xyloglucan. These results indicate that the cross-links are made of xyloglucan. We discussed the possibility of cross-links involved in the control of mechanical properties of the cell wall. PMID- 10845463 TI - Molecular and biochemical characterization of a novel hydroxycinnamoyl-CoA: anthocyanin 3-O-glucoside-6"-O-acyltransferase from Perilla frutescens. AB - We have isolated and characterized a cDNA, PfAT208, encoding hydroxycinnamoyl CoA: anthocyanin 3-O-glucoside-6"-O-acyltransferase (3AT) from Perilla frutescens. The identity of the cDNA was established by determination of the reaction products with recombinant enzyme overexpressed in Escherichia coli. The deduced amino acid sequence has a few regions that are conserved in a CoA dependent acyltransferase family. The recombinant enzyme produced in yeast could utilize cyanidin 3-glucoside and cyanidin 3,5-diglucoside, putative substrates in vivo, as well as other anthocyanins. The inhibitory effects of diethyl pyrocarbonate and N-ethylmaleimide on the recombinant 3AT activities suggest that histidine and cysteine residues are important for their catalytic function. These properties are in common with anthocyanin 5-O-glucoside-6"-O-acyltransferase (5AT). In Northern analysis, a transcript of PfAT208 was detected in the young leaves of perilla red forma. The properties of other cDNAs, gentian GAT106 and petunia PhAT48, isolated during the above cloning procedure are also described. PMID- 10845464 TI - Isolation and characterization of a 60 kDa 2,4-D-binding protein from the shoot apices of peach trees (Prunus persica L.); it is a homologue of protein disulfide isomerase. AB - To obtain a candidate auxin-binding protein (ABP), a soluble 60 kDa protein was isolated from an extract of shoot apices of peach trees (Prunus persica L.) by affinity chromatography on a 2,4-dichlorophenoxyacetic acid (2,4-D)-linked Sepharose4B column. The 60 kDa polypeptide, designated Pp60, was purified as a single band on SDS-PAGE by column chromatography. Its dissociation constant (Kd) for [14C]-2,4-D was calculated to be 3.5 x 10(-5) M. The binding of Pp60 for [14C]-2,4-D was inhibited by naphthalene-1-acetic acid (NAA) and p chlorophenoxyisobutyric acid (PCIB) as well as 2,4-D. Indole-3-acetic acid (IAA) had little effect on the binding. These results suggested that Pp60 is a protein that has an affinity for 2,4-D, NAA, and PCIB in vitro. The partial amino acid sequences of Pp60 showed high homology to those of protein disulfide isomerase (EC 5.3.4.1). Immunoblot analysis demonstrated that Pp60 exists ubiquitously in shoots and leaves. In fruit, expression of Pp60 is restricted at an early stage of development. PMID- 10845465 TI - Changes in the apoplastic pH are involved in regulation of xyloglucan breakdown of azuki bean epicotyls under hypergravity conditions. AB - Hypergravity inhibited elongation growth of azuki bean (Vigna angularis Ohwi et Ohashi) epicotyls by decreasing the mechanical extensibility of cell walls via the increase in the molecular mass of xyloglucans [Soga et al. (1999) Plant Cell Physiol. 40: 581]. Here, we report that the pH value of the apoplastic fluid in epicotyls increased from 5.8 to 6.6 by hypergravity (300 x g) treatment. When the xyloglucan-degrading enzymes extracted from cell walls of the 1 x g control epicotyls were assayed in buffer at pH 6.6 and 5.8, the activity at pH 6.6 was almost half of that at pH 5.8. In addition, when enzymically active cell wall preparations obtained from 1 x g control epicotyls were autolyzed in buffer at pH 5.8 and 6.6 and then xyloglucans were extracted from the autolyzed cell walls, the molecular mass of xyloglucans incubated at pH 5.8 decreased during the autolysis, while that at pH 6.6 did not change. Thus, the xyloglucans were not depolymerized by autolysis at the pH value (6.6) observed in the hypergravity treated epicotyls. These findings suggest that in azuki bean epicotyls, hypergravity decreases the activities of xyloglucan-degrading enzymes by increasing the pH in the apoplastic fluid, which may be involved in the processes of the increase in the molecular mass of xyloglucans, leading to the decrease in the cell wall extensibility. PMID- 10845466 TI - Effects of high-temperature treatments on a thermophilic cyanobacterium Synechococcus vulcanus. AB - Effects of high-temperature treatments on a thermophilic cyanobacterium, Synechococcus vulcanus, were studied, and the following results were obtained. (1) Oxygen evolution and the PSII photochemical reaction were the most sensitive sites and started to be inactivated at temperatures slightly higher than the cultivating temperature. (2) The decrease in the fluorescence Fv value reflected the inactivation of the charge separation reaction of PSII as well as that of the oxygen evolution reaction. (3) The dark fluorescence level, Fo, showed an increase at around 70 degrees C, which was partially reversed by further incubation at 50 degrees C. This increase reflected the inactivation of PSII reaction centers and probably dissociation of phycobilisomes from the PSII reaction center complexes. (4) At higher temperatures, phycobiliproteins disassembled and denatured in a pH-dependent manner, causing a large Fo decrease. (5) Cell membranes became leaky to low-molecular-weight substances at around 72 degrees C. (6) Inhibition of growth of the cells was recognized when the cells were pretreated at temperatures higher than 72 degrees C. Reversibility of the high-temperature effects and relationship between viability of the cells and the degradation of the cell membranes are discussed. PMID- 10845467 TI - Modulation of 14-3-3 protein interactions with target polypeptides by physical and metabolic effectors. AB - The proteins commonly referred to as 14-3-3s have recently come to prominence in the study of protein:protein interactions, having been shown to act as allosteric or steric regulators and possibly scaffolds. The binding of 14-3-3 proteins to the regulatory phosphorylation site of nitrate reductase (NR) was studied in real time by surface plasmon resonance, using primarily an immobilized synthetic phosphopeptide based on spinach NR-Ser543. Both plant and yeast 14-3-3 proteins were shown to bind the immobilized peptide ligand in a Mg2+-stimulated manner. Stimulation resulted from a reduction in KD and an increase in steady-state binding level (Req). As shown previously for plant 14-3-3s, fluorescent probes also indicated that yeast BMH2 interacted directly with cations, which bind and affect surface hydrophobicity. Binding of 14-3-3s to the phosphopeptide ligand occurred in the absence of divalent cations when the pH was reduced below neutral, and the basis for enhanced binding was a reduction in K(D). At pH 7.5 (+Mg2+), AMP inhibited binding of plant 14-3-3s to the NR based peptide ligand. The binding of AMP to 14-3-3s was directly demonstrated by equilibrium dialysis (plant), and from the observation that recombinant plant 14-3-3s have a low, but detectable, AMP phosphatase activity. PMID- 10845468 TI - Oxygen uptake, acidification of medium and nitrate uptake induced by blue light in nitrate-starved Chlorella cells. AB - Blue light-induced oxygen uptake of the colorless mutant of Chlorella kessleri (No. 9.80) was 30-40% higher in the presence of exogenous glycine than in its absence. None of the other amino acids tested had this effect. Moreover, mutant cells in which glutamine synthetase was inhibited by methionine sulphoximine, accumulated approximately 65% more ammonium ions under blue irradiation in the presence of exogenous glycine than in its absence. The protein kinase C inhibitors, staurosporine or K252a, reduced the enhancement of oxygen uptake by approximately 40%. The present results indicate that blue light-dependent deamination of endogenous glycine might be a prerequisite for enhanced oxygen uptake in Chlorella. This blue light-induced oxygen uptake was not influenced by the inhibitors of protein phosphatase, calyculin A or okadaic acid. On the contrary, calyculin A and okadaic acid had a marked effect on the acidification of the suspension medium and nitrate uptake induced by blue light in Chlorella cells. The different responses to the inhibitors of protein kinase and phosphatase suggest the presence of different pathways among the blue light signal transduction operating on oxygen uptake, acidification of the medium and nitrate uptake in Chlorella. PMID- 10845469 TI - Pulmonary embolism: what's wrong with this diagnosis? PMID- 10845470 TI - Nonthrombotic pulmonary emboli. PMID- 10845471 TI - Progress and complexity for the next century. PMID- 10845472 TI - Pitfalls of the vague radiology report. PMID- 10845473 TI - The virtual course: delivery of live and recorded continuing medical education material over the Internet. AB - OBJECTIVE: Our objective was to deliver live and recorded lectures from a continuing medical education course, including the representations of original slides, over the Internet, using streaming audio media. CONCLUSION: The streaming audio media, an emerging technology, not only delivers large lectures over the Internet using commercially available PCs and modems, but also allows review at any time by individuals with access to the Web. An interactive conference of continuing medical education curriculum can thus be delivered and continuing medical education credit earned without leaving home. PMID- 10845474 TI - Practical business aspects of radiology. PMID- 10845475 TI - Double-contrast barium enema for colorectal cancer screening: a review of the issues and a comparison with other screening alternatives. PMID- 10845476 TI - Intestinal perforation from blunt trauma to an inguinal hernia. PMID- 10845477 TI - CT colonography for colorectal cancer screening: lessons from mammography. PMID- 10845478 TI - Comparison of time-efficient CT colonography with two- and three-dimensional colonic evaluation for detecting colorectal polyps. AB - OBJECTIVE: We compared the findings of time-efficient CT colonography with complete two-dimensional (2D) and three-dimensional (3D) CT colonography and conventional colonoscopy in detecting colorectal polyps. SUBJECTS AND METHODS: Forty-two patients undergoing colonoscopy screening were examined with CT colonography before endoscopy. Data were examined following one of two methods. In method 1, axial 2D data sets were examined in a cine mode. If findings were suggestive of abnormality, focal areas were examined with 3D CT colonography. In method 2, data sets were examined exactly as in method 1, and subsequent to that review, data sets were examined with simultaneous 3D "fly-through" CT colonography (surface-rendered images) and multiplanar reformatted images. The time required to examine CT colonography using each technique was recorded and abnormal findings were documented. Results of methods 1 and 2 were compared with findings on colonoscopy. RESULTS: Colonoscopy detected 16 polyps in 13 patients (polyp size, 2-10 mm). Ten polyps measured 5 mm or less, five measured between 6 and 9 mm, and one measured 10 mm or more. Using method 1, two of 10 polyps measuring less than 5 mm, three of five polyps measuring between 6 and 9 mm, and one polyp measuring 10 mm were detected. We noted no false-positive polyps. Average evaluation time was 16 min. With method 2, the same polyps were seen as with method 1. No additional polyps were detected, and the average evaluation time was 40 min. CONCLUSION: Axial 2D CT colonography can be performed quickly and is comparable with complete 2D and 3D CT colonography in detecting colorectal polyps. PMID- 10845479 TI - Comparing MR imaging and CT in the staging of gastric carcinoma. AB - OBJECTIVE: The purpose of this study was to assess the usefulness of breath-hold two-dimensional (2D) fast low-angle shot (FLASH) and T2-weighted turbo spin-echo fast MR imaging compared with helical CT in the staging of gastric carcinoma. SUBJECTS AND METHODS: Thirty patients with gastric carcinoma underwent preoperative MR imaging and helical CT. MR imaging at 1.5 T was performed immediately after the intramuscular injection of scopolamine and the oral administration of water or effervescent granules. Breath-hold 2D FLASH T1 weighted images in all three planes, turbo spin-echo T2-weighted axial images, and gadolinium-enhanced fat-suppressed 2D FLASH axial images were included. Helical CT was performed 60 sec after initiation of i.v. contrast medium injection (2.5-3 ml/sec). Two groups of two radiologists each independently analyzed the MR and helical CT findings, and these results were compared with the pathologic findings. RESULTS: For T staging, MR imaging accuracy was higher than that of helical CT (73.3% and 66.7%, respectively); however, the accuracies of the two methods were not significantly different from each other (McNemar test, p > 0.05). Overstaging was noted in 6.7% of cases with MR imaging and 10% with helical CT. Understaging was noted in 20% of cases with MR imaging and 23.3% with helical CT. For N staging, the accuracies of MR imaging and helical CT were 55% and 58.6%, respectively, with no statistical significance (overstaging, 10% and 6.9%; understaging, 34.5% and 34.5%, respectively). CONCLUSION: MR imaging was comparable to helical CT in the T and N staging of gastric cancer. PMID- 10845480 TI - Imaging of peritoneal inclusion cysts. PMID- 10845481 TI - Early duodenal cancer: detection on double-contrast upper gastrointestinal radiography. PMID- 10845482 TI - Hemodynamics of small hepatic focal nodular hyperplasia: evaluation with single level dynamic CT during hepatic arteriography. AB - OBJECTIVE: We examined the usefulness of single-level dynamic CT during hepatic arteriography to observe the hemodynamics of small hepatic focal nodular hyperplasia. CONCLUSION: Single-level dynamic CT during hepatic arteriography revealed not only centrifugal blood supply through the fibrous stellate scar, but also the drainage to dilated veins in or near the focal nodular hyperplasia nodule and directly to the hepatic sinusoid in the surrounding liver. PMID- 10845483 TI - Atypical inside-out pattern of hepatic hemangiomas. PMID- 10845485 TI - Sonography of an arteriovenous-type hemangioma of the testis. PMID- 10845484 TI - Initial experience with contrast-enhanced sonography of the prostate. AB - OBJECTIVE: We investigated the usefulness of contrast-enhanced sonography to depict vascularity in the prostate and improve the detection of prostatic cancer. SUBJECTS AND METHODS: Twenty-six patients with an elevated prostate-specific antigen level (> or = 4 ng/ml) or an abnormal digital rectal examination were enrolled in a phase II study of an i.v. injected sonographic contrast agent. Continuous gray-scale, intermittent gray-scale, phase inversion gray-scale, and power Doppler sonography of the prostate were performed. Sonographic findings were correlated with sextant biopsy results. RESULTS: After the administration of contrast material, gray-scale and Doppler images revealed visible enhancement (p < 0.05). Using intermittent imaging, we found focal enhancement in two isoechoic tumors that were not visible on baseline images. No definite focal area of enhancement was identified in any patient without cancer. Contrast-enhanced images revealed transient hemorrhage in the biopsy tracts of three patients. CONCLUSION: Enhancement of the prostate can be seen on gray-scale and Doppler sonographic images after the administration of an i.v. contrast agent. Contrast enhanced intermittent sonography of the prostate may be useful for the selective enhancement of malignant prostatic tissue. PMID- 10845486 TI - Time-resolved contrast-enhanced MR angiography of renal artery stenosis: diagnostic accuracy and interobserver variability. AB - OBJECTIVE: The purpose of this study was to evaluate diagnostic accuracy and interobserver variability of time-resolved three-dimensional gadolinium-enhanced MR angiography in the detection of renal artery stenosis in comparison with intraarterial digital subtraction angiography as the standard of reference. SUBJECTS AND METHODS: Forty consecutive patients (age range, 25-81 years; mean, 62.9 +/- 11.9 years) with suspected renal artery stenosis underwent intraarterial digital subtraction angiography and gadolinium-enhanced MR angiography, performed on a 1.5-T system with fast low-angle shot three-dimensional imaging (3.8/1.49 [TR/TE], 25 degrees flip angle, 10-sec acquisition time, and 1.5-mm partition thickness). Three time-resolved phases were obtained in a single breath-hold. Digital subtraction angiography and gadolinium-enhanced MR angiography were evaluated by four observers who studied 80 main renal arteries and 19 accessory vessels to evaluate the degree of stenosis. A stenosis reducing the intraarterial diameter by more than 50% was regarded as hemodynamically significant. Interobserver variability was calculated. RESULTS: Only one gadolinium-enhanced MR angiography study was not of diagnostic quality, as a result of failure of the power injector. All main branches were of diagnostic quality in 38 (97.4%) of the remaining 39 gadolinium-enhanced MR angiography studies. Seventeen (89.5%) of 19 accessory renal arteries were depicted with gadolinium-enhanced MR angiography. The overall sensitivity for significant stenoses was 92.9%. The overall specificity was 83.4%, and the overall accuracy was 85.9%. Interobserver variability of gadolinium-enhanced MR angiography exceeded that of digital subtraction angiography. CONCLUSION: Time-resolved three-dimensional gadolinium enhanced MR angiography is a useful noninvasive method of screening suspected renal artery stenosis because of its easy application, short examination time, and high sensitivity despite of its higher interobserver variability. PMID- 10845487 TI - Hysterosonographically guided endometrial biopsy: technical feasibility. PMID- 10845488 TI - Combined embolization and percutaneous radiofrequency ablation of a solid renal tumor. PMID- 10845489 TI - Sonographic guidance of laparoscopic renal cryoablation. PMID- 10845490 TI - Wegener's granulomatosis presenting as a renal mass. PMID- 10845491 TI - Ultrafast MR imaging of the fetus. AB - OBJECTIVE: We examined the capability of ultrafast single-shot fast spin-echo imaging to assess different fetal organ systems compared with prenatal sonography, using autopsy or postpartum imaging as a standard of reference. SUBJECTS AND METHODS: Thirty women with complicated pregnancies (mean age of gestation, 190 +/- 54 days) underwent T2-weighted ultrafast MR imaging. MR images were analyzed with regard to diagnostic confidence in assessing abnormalities of fetal organ systems, and data were correlated with postpartum findings or necropsy. Results were compared with those of prenatal sonography. RESULTS: Using receiver operating characteristic curve analysis, diagnostic confidence of MR imaging was best for assessing the brain (area under the curve [Az] = 0.96) and spinal canal (Az = 1.0), uteroplacental unit (Az = 0.93), and lungs (Az = 0.91). Results for the heart (Az = 0.63) and extremities (Az = 0.77) were significantly lower than that of other organs (p < 0.001). Diagnostic accuracy increased with gestational age. No statistically significant difference between sonography and MR imaging was found for the detection of abnormality in any organ system. In three fetuses, MR imaging was superior to sonography in characterizing cerebral abnormalities. MR imaging was inferior to sonography in characterizing abnormalities of the heart and extremities. CONCLUSION: Our results indicate that ultrafast MR imaging can be used for in vivo fetal imaging, especially in assessing cerebral abnormalities. However, MR imaging should be restricted to situations in which sonographic findings are ambiguous or impaired. PMID- 10845492 TI - Prenatal MR imaging of congenital diaphragmatic hernia. PMID- 10845493 TI - Abdominal sonography in examination of children with blunt abdominal trauma. AB - OBJECTIVE: The objective of our study was to evaluate abdominal sonography for the detection of fluid and organ injury in children with blunt abdominal trauma. SUBJECTS AND METHODS: Fifty-one consecutive children with blunt abdominal trauma requiring abdominal CT were prospectively examined with sonography. Sonograms and CTs were independently evaluated by two radiologists for fluid and organ injury; CT examinations were considered abnormal if either was identified. Differences in CT interpretation were settled by a third observer. Using CT as the truth standard, we calculated the sensitivity, specificity, and negative predictive value of sonography for both observers. Agreement of the sonographic interpretations was evaluated using kappa statistic. RESULTS: In 33.3% of patients, CT revealed fluid, organ injury, or both. The sensitivity and specificity of sonography when detection of fluid was the sole parameter evaluated was 58.8% and 79.4%, respectively, for observer 1 and 47.1% and 79.4%, respectively, for observer 2. In contrast, the sensitivity and specificity of sonography when detection of both fluid and organ injury was evaluated was 64.7% and 79.4%, respectively, for observer 1 and 70.6% and 70.6%, respectively, for observer 2. The negative predictive value of sonography was 79.4% and 75.0% with evaluation limited to detection of fluid and 81.8% and 82.8% with evaluation of fluid and organ abnormality for observers 1 and 2 , respectively. Agreement was excellent for sonographic identification of fluid (kappa = 0.82) but poor for detection of organ injury (kappa = 0.34). CONCLUSION: The low sensitivity and negative predictive value of sonography when assessing for either fluid alone or fluid and organ injury suggest that a normal screening sonography alone in the setting of blunt abdominal trauma fails to confidently exclude the presence of an intraabdominal injury. PMID- 10845494 TI - Radiography of cervical spine injury in children: are flexion-extension radiographs useful for acute trauma? AB - OBJECTIVE: We assessed the role of cervical spine flexion-extension radiographs in the acute evaluation of pediatric trauma patients. MATERIAL AND METHODS: We performed a retrospective review of all pediatric trauma patients who underwent static cervical spine radiography followed by flexion-extension radiography during a 22-month period. We reviewed the mechanism of injury, physical examination findings, and patient age, and tabulated the results of initial and follow-up imaging studies. RESULTS: Two hundred forty seven children (age range, 1.6-18 years; mean age, 11.5 years) with a history of trauma underwent cervical spine radiography followed by flexion-extension radiography. Static cervical spine radiographs revealed normal findings in 224 patients (91%). Flexion extension radiographs revealed normal findings for all patients with normal findings on cervical spine radiographs. Of 23 children (9%) with abnormal findings on static cervical spine radiographs, seven (30%) had congenital abnormalities visible on flexion-extension radiographs; 10 (43%) had traumatic injuries including fracture, subluxation, or soft-tissue swelling; two (9%) had instability; and six (26%) had questionable abnormalities that were noted on static cervical spine radiographs. In four patients (66%) with abnormal findings on static cervical spine radiographs, flexion-extension radiographs were helpful in ruling out abnormality. CONCLUSION: In children with a history of trauma and normal findings on static cervical spine radiographs, additional flexion extension radiographs are of questionable use. PMID- 10845495 TI - Multislice helical CT to facilitate combined CT of the neck, chest, abdomen, and pelvis in children. PMID- 10845496 TI - MR imaging differentiation of soft-tissue hemangiomas from malignant soft-tissue masses. AB - OBJECTIVE: The purpose of this study was to determine whether MR imaging features can reliably distinguish hemangiomas from malignant soft-tissue masses. MATERIALS AND METHODS: We retrospectively reviewed MR imaging studies of 22 patients with soft-tissue hemangiomas and 22 patients with malignant soft-tissue masses. Images were reviewed and agreement reached by a consensus interpretation of two observers and by an independent observer. Masses were evaluated for signal intensity on T1- and T2-weighted images, for enhancement with gadolinium administration, and for morphology (lobulation, septation, central low-intensity dots). Lesion T2 signal and lesion enhancement with gadolinium administration were also objectively measured using regions of interest and comparison with skeletal muscle. RESULTS: Signal intensity on T1-weighted imaging of hemangiomas and malignant soft-tissue masses was similar. Subjective analysis showed greater T2 signal and gadolinium enhancement in hemangiomas; however, the differences were not statistically significant on objective analysis. Lobulation, septation, and central low-signal-intensity dots were all more common in hemangiomas, with statistical significance achieved; the combination of all three findings was specific for hemangioma. CONCLUSION: Although no single MR imaging feature was diagnostic in this study, analysis of lesion morphology, signal intensity, and enhancement with gadolinium allowed MR imaging differentiation of hemangiomas from malignant soft-tissue masses. PMID- 10845497 TI - Doppler sonography of the anterior ascending cervical arteries of the hip: evaluation of healthy and painful hips in children. AB - OBJECTIVE: We evaluated the Doppler sonography of small feeding arteries to the femoral head in children. SUBJECTS AND METHODS: In a prospective study of 224 hips in 112 patients (mean age, 5 years 11 months), the anterior ascending cervical arteries of the hip were identified with color Doppler sonography. Subsequently, we measured the resistive index (RI) with pulsed Doppler sonography. RESULTS: In 61% (137/224) of hips, a Doppler signal could be obtained. In asymptomatic hips (n = 64), the mean RI was 0.58. In symptomatic hips, the definitive diagnoses and mean values of RI included transient synovitis (n = 31) and 0.92, Perthes' disease (n = 9) and 0.67, and miscellaneous (n = 5) and 0.68. In 28 symptomatic hips, no definite diagnosis could be determined and the complaints spontaneously disappeared during follow-up (mean RI, 0.57). We found no statistically significant difference in the RI of symptomatic versus asymptomatic hips, except in patients with transient synovitis (p < 0.001). In 11 hips with transient synovitis that were reexamined after 4-6 weeks, the RI returned to normal (0.57). The RI in symptomatic hips showed a positive correlation with the amount of effusion (r = 0.69, p < 0.001). In symptomatic and asymptomatic hips, we found no correlation with age (p = 0.9 and 0.1, respectively). CONCLUSION: The deep capsular vessels of the hip joint can be evaluated on Doppler sonography in more than 60% of hips. Also, the RI is age independent and correlates with the amount of effusion. PMID- 10845498 TI - Dynamic MR imaging of the hip in Legg-Calve-Perthes disease: comparison with arthrography. PMID- 10845499 TI - Sonography and CT of pancreatoblastoma in children. AB - OBJECTIVE: We reveal the sonography and CT findings of three children with pancreatoblastoma. CONCLUSION: Pancreatoblastomas are invasive tumors that encase adjacent vessels and infiltrate surrounding organs. Original and atypical findings include metastases to the omentum and lymph nodes of the neck and direct extension of the tumor to the portal vein and its branches. PMID- 10845500 TI - Quantitative comparison of intrabrain diffusion in adults and preterm and term neonates and infants. AB - OBJECTIVE: Quantitative measurements of mean water diffusivity (D(av)) were made in human neonates, infants, and adults to assess changes in brain tissue that occur with maturation. SUBJECTS AND METHODS: Values of D(av) were obtained by calculating the average of the diffusion measurements made with diffusion sensitizing gradients placed along three orthogonal directions. The mean diffusivity, a rotationally invariant determination of apparent diffusion coefficient, was measured in five healthy prematurely born neonates and infants, in 10 healthy term neonates and infants, and in five adults. RESULTS: Values of D(av) were found to decrease with maturation in most parts of the brain. In prematurely born neonates and infants with a postmenstrual age (postgestastional age + postnatal age) under 36 weeks, the average value of D(av) in frontal white matter was 1.90 x 10(-3) mm2 sec(-1). The corresponding value was measured as 1.62 x 10(-3) mm2 sec(-1) in neonates and infants born at term with a postnatal age of no more than 43 days and 0.79 x 10(-3) mm2 sec(-1) in the adult brain. CONCLUSION: Values of D(av) are known to decrease in neonates and young infants in the period immediately after ischemic insult. This decrease and the associated increase in signal intensity seen on diffusion-weighted imaging have been used to monitor ischemic brain injury in neonates and infants. Therefore, the decrease in D(av) that occurs with maturation, which we report in this study, must be considered if quantitative diffusion measurements are used to assess ischemic neonatal brain injury. PMID- 10845501 TI - MR imaging of herpes simplex type 1 encephalitis in infants and young children: a separate pattern of findings. AB - OBJECTIVE: We sought to identify the initial MR findings of herpes simplex encephalitis in infants and young children. CONCLUSION: MR imaging findings of herpes encephalitis in infants and young children appear to differ from those seen in neonates, older children, and adults. Appreciation of this MR imaging pattern coupled with a strong clinical suspicion of herpes helps to ensure the correct diagnosis is made. PMID- 10845502 TI - Wall shear stress and early atherosclerosis: a review. PMID- 10845503 TI - Helical CT angiography of thoracic outlet syndrome: functional anatomy. AB - OBJECTIVE: The objective of this study is to determine the anatomic characteristics of the thoracic outlet in symptomatic patients before and after postural maneuver. SUBJECTS AND METHODS: Seventy-nine symptomatic patients (61 female patients [group 1]; 18 male patients [group 2]; mean age, 38 years) underwent helical CT angiography of the thoracic apices in the neutral position and after a postural maneuver, enabling the evaluation of the functional anatomy of the musculoskeletal and arterial structures of the ipsilateral thoracic outlet. RESULTS: A statistically significant difference was found between the distribution of the distances (maximum and costosubclavian) measured in the neutral position and after postural maneuver in groups 1 and 2. The median value of these distances was smaller after postural maneuver in groups 1 and 2. A statistically significant difference was found between the distribution of the distances (maximum and costosubclavian) measured in patients of group 1 with arterial stenosis and in patients of group 1 without arterial stenosis. A slight indentation of the anterior wall of the subclavian artery when it arches around the anterior scalene muscle was observed in 39 patients (64%) in group 1 and in 11 patients (61%) in group 2 in the neutral position, in 19 patients (31%) in group 1 and in six patients (33%) in group 2 after the postural maneuver. The predominant positional changes of the vascular structures were the posteroanterior displacement of the subclavian vessels observed in groups 1 and 2, the arch made by the subclavian artery above the first rib in 40 patients (66%) in group 1 and nine patients (50%) in group 2, and the posterior displacement of the axillary artery observed in 36 patients (59%) in group 1 and in 12 patients (67%) in group 2. CONCLUSION: Helical CT shows significant narrowing of the costoclavicular space after postural maneuver in symptomatic patients. PMID- 10845504 TI - Radiologic findings of abdominal polyarteritis nodosa. AB - OBJECTIVE: The purpose of this study was to evaluate the radiographic and imaging findings of seven patients with polyarteritis nodosa involving the abdomen. CONCLUSION: Vasculitis should be considered when multiple abdominal organs have abnormal radiologic findings; however, angiography is necessary to specifically diagnose polyarteritis nodosa. PMID- 10845505 TI - Intracavitary urokinase for enhancement of percutaneous abscess drainage: phase II trial. AB - OBJECTIVE: To evaluate the effectiveness of urokinase as an abscess-cavity irrigant during percutaneous abscess drainage. SUBJECTS AND METHODS: In a prospective study, approved by the Food and Drug Administration and the review board at our institution, urokinase and saline were used as abscess-cavity irrigants. In the study group of 42 patients, half the patients were randomly placed into the urokinase group and the other half were placed into the control saline group. Doses used varied with the size of the abscess. Data collected from patient charts were evaluated with standard statistical methods. RESULTS: The results indicate definite benefits of the urokinase treatment. The length of stay (p = 0.0025) and treatment costs (p = 0.0021) were significantly less for the urokinase group. Other clinical parameters, including the febrile course, elevated WBC, and days of drainage, trended in a favorable fashion. CONCLUSION: Urokinase injected intracavitarily is an effective technique for shortening the treatment time and improves the clinical course for patients treated with percutaneous drainage techniques. PMID- 10845506 TI - A simple device for continuous leg extension during CT-guided interventions. PMID- 10845507 TI - Needle localization using a stereotactic table: a reassessment. PMID- 10845508 TI - Contribution of individual projections alone and in combination for radiographic detection of ankle fractures. AB - OBJECTIVE: We wanted to determine whether the standard three-view ankle radiographic series could be replaced by a two-view combination, and if so, which two-view combination (anteroposterior with lateral or mortise with lateral) would be superior. MATERIALS AND METHODS: During a 12-month period, we retrospectively reviewed 556 consecutive ankle radiographic studies consisting of anteroposterior, mortise, and lateral views. One hundred twenty patients with at least one ankle fracture were paired with 140 healthy control subjects. Each image in the three-view examination was separated and sorted by view and studied independently; all images were reviewed by two skeletal radiologists and two orthopedic surgeons. Each radiograph was evaluated for fracture of the medial, lateral, and posterior malleoli and the foot using a five-point confidence rating. Performance of each view and modeled two- and three-view combinations of views was evaluated with modified receiver operating characteristic analysis. RESULTS: The data provide little support for preferring either two-view combination (anteroposterior-lateral or mortise-lateral) for any type of fracture. The three-view combination does detect significantly more fractures than some two-view combinations in some locations, and there is a statistically significant cost in diagnostic accuracy for eliminating the anteroposterior or mortise view. CONCLUSION: Reducing the ankle radiographic series from three to two views would result in a small but significant decrease in the detection of fractures of the ankle and foot. Both two-view combinations are equivalent for fracture detection. PMID- 10845509 TI - Parosteal lipoma of the fibula. PMID- 10845510 TI - Dual X-ray absorptiometry: recognizing image artifacts and pathology. PMID- 10845511 TI - MR imaging and MR arthrography of paraglenoid labral cysts. AB - OBJECTIVE: We investigated the pathophysiology of paraglenoid labral cysts on the basis of MR imaging, MR arthrography, and cyst aspiration. MATERIALS AND METHODS: From 2211 MR imaging examinations, 51 (2.3%) cysts in 46 patients were identified. MR arthrography (n = 5), cystography (n = 1), arthroscopy (n = 17), percutaneous needle aspiration (n = 4), and medical records were also reviewed (n = 46). RESULTS: On MR imaging and arthrography, cysts were best viewed on T2 weighted images. Mean cyst diameter and volume were 2.2 cm and 2.8 cm3, respectively. Fifty-seven percent of cysts were located adjacent to the posterior labrum. On MR imaging and arthroscopy, a labral tear was identified in 27 (53%) and 15 (88%) patients, respectively. Eight cysts that caused compression neuropathy were large (mean size, 3.1 cm; p = 0.04) and located next to the posterior or inferior labrum. In four of five patients, MR arthrograms showed no intraarticular contrast material in the cyst. Cystograms showed no communication with the glenohumeral joint space, and cyst aspiration resulted in temporary symptom relief; however, cysts recurred in three of four patients. CONCLUSION: Most paralabral cysts are associated with labral tears. Paralabral cysts may be difficult to identify on MR arthrography unless a T2-weighted sequence is performed. Direct communication between a cyst and joint space rarely occurs. A posterior or inferior cyst may cause compression neuropathy of the suprascapular or axillary nerve, respectively. Cyst aspiration may result in temporary relief of symptoms, but an untreated labral tear should be suspected if cysts recur. PMID- 10845513 TI - Sonography of Morton's neuromas. AB - OBJECTIVE: The objective is to evaluate the sonographic characteristics of Morton's neuromas and the usefulness of sonography in detecting them. MATERIALS AND METHODS: Thirty intermetatarsal spaces were evaluated with sonography to diagnose Morton's neuromas. The prospective sonographic reports were correlated with surgical and pathologic findings. Each sonogram was then blindly and retrospectively reviewed to characterize mass echotexture, location, size, and continuity with the plantar digital nerve when present. RESULTS: Surgery revealed 27 Morton's neuromas, one synovial cyst with infarcted tissue, one ganglion cyst, and one giant cell tumor of the tendon sheath. The prospective sonographic reports correctly identified neuromas in 85% of the cases. Retrospectively, 79.2% (19/24) of the neuromas were characterized as hypoechoic compared with muscle, whereas 12.5% (3/24) were of mixed echotexture and 8.3% (2/24) were anechoic. One half (50%) of the neuromas were located dorsal to the plantar aspect of the metatarsal heads, and 50% were both dorsal and plantar to this level. No statistical difference in height and width was found between neuromas and nonneuromas; however, nonneuromas were statistically greater in length than neuromas. All 15 masses in which presumed plantar digital nerve continuity with the mass was identified were neuromas. CONCLUSION: Sonography can reveal a Morton's neuroma in 85% of cases. Identification of the presumed plantar digital nerve in continuity with the mass improves diagnostic confidence. The finding of an interdigital mass greater than 20 mm in length should raise suspicion of an abnormality other than a neuroma. PMID- 10845512 TI - Sonography of the glenoid labrum: a cadaveric study with arthroscopic correlation. AB - OBJECTIVE: We assessed the usefulness of sonography in evaluating the glenoid labrum in cadaveric specimens using arthroscopy as a standard of reference. MATERIALS AND METHODS: Eighty labral quadrants in 20 cadaveric shoulders were examined by two musculoskeletal radiologists using 5- to 7-MHz linear and curvilinear transducers. Agreement was reached by consensus. After sonography, arthroscopy was performed by an experienced orthopedic surgeon. Each labral quadrant was classified at the time of sonography and arthroscopy as normal, degenerated, or torn. RESULTS: Concordance between sonography and arthroscopy was 86% (69/80 quadrants). In differentiating abnormal labrum (tear or degeneration) from normal labrum using sonography, sensitivity was 63%, specificity was 98%, positive predictive value was 94%, negative predictive value was 86%, and accuracy was 88%. In differentiating labral tears from other labral conditions (degeneration or normality), sensitivity was 67%, specificity was 99%, positive predictive value was 67%, negative predictive value was 99%, and accuracy was 98%. CONCLUSION: Sonography has a promising role in the evaluation of the glenoid labrum, particularly in excluding labral tears when the labra appear normal on sonography. Further studies are required using normal and symptomatic patients to determine the usefulness of sonography in the diagnosis of labral abnormalities. PMID- 10845514 TI - Neoplasms of the scapula. PMID- 10845515 TI - Analysis of first-pass and delayed contrast-enhancement patterns of dysfunctional myocardium on MR imaging: use in the prediction of myocardial viability. AB - OBJECTIVE: The purpose of the study was to analyze first-pass and delayed contrast-enhancement patterns of dysfunctional myocardial regions on MR imaging after injection of gadopentetate dimeglumine to predict myocardial viability in patients with coronary artery disease. SUBJECTS AND METHODS: Twelve patients with wall motion abnormalities and related coronary artery disease revealed by conventional coronary angiography underwent MR imaging at 1.5-T before and 3 months after revascularization therapy. Short-axis images were acquired using a cine gradient-echo sequence. Each slice was divided into eight segments. Overall, 73 segments with impaired contractility were imaged during the first-pass and 14 +/- 2 min after injection of 0.05-mmol/kg gadopentetate dimeglumine at a flow of 3 ml/sec using a T1-weighted turbo fast low-angle shot sequence. Improved systolic wall thickening 3 months after revascularization served as the criterion of viability. RESULTS: At study entry, 26 dysfunctional segments showed delayed hyperenhancement compared with the adjacent functional segments within the same slice, and 47 did not reveal hyperenhancement. After revascularization, 25 (96%) of the 26 hyperenhanced segments did not recover function, whereas 39 (83%) of the 47 segments without hyperenhancement showed mechanical improvement. Segment related sensitivity and specificity for the correlation of lack of delayed hyperenhancement with myocardial viability were 39 (98%) of 40 and 25 (76%) of 33, respectively. Hypoenhancement during first-pass did not serve as a reliable criterion of viability. CONCLUSION: Evidence of delayed hyperenhancement of dysfunctional myocardium may be used to predict lack of mechanical improvement or nonviability, whereas the lack of hyperenhancement can be correlated with improvement of regional contractility or viability after revascularization. PMID- 10845516 TI - Myocardial viability using MR imaging: is it ready for clinical use? PMID- 10845517 TI - Acute parenchymal lung disease in immunocompetent patients: diagnostic accuracy of high-resolution CT. AB - OBJECTIVE: The purpose of this study was to determine whether acute parenchymal lung diseases can be differentiated on the basis of the pattern and distribution of abnormalities revealed on high-resolution CT. MATERIALS AND METHODS: High resolution CT scans of 90 patients with acute parenchymal lung diseases (19 with bacterial pneumonia, 13 with mycoplasmal pneumonia, 21 with acute interstitial pneumonia, 18 with hypersensitivity pneumonitis, 10 with acute eosinophilic pneumonia, and nine with pulmonary hemorrhage) were independently assessed by two observers who had no knowledge of clinical or pathologic data. The observers recorded abnormalities, their first-choice diagnosis, and their degree of confidence in their first-choice diagnosis. RESULTS: The two observers made a correct first-choice diagnosis in an average of 55 (61%) of 90 cases. Correct first-choice diagnosis was made in 50% of cases of bacterial pneumonia, 62% of mycoplasmal pneumonia, 90% of acute interstitial pneumonia, 72% of hypersensitivity pneumonitis, 30% of acute eosinophilic pneumonia, and 28% of pulmonary hemorrhage. CT findings allowed distinction between infectious and noninfectious causes in 81 (90%) of 90 cases. CONCLUSION: High-resolution CT is helpful in the differential diagnosis of infectious from noninfectious acute parenchymal lung disease. However, high-resolution CT is of limited value in making a specific diagnosis. PMID- 10845518 TI - Sarcoidosis with pulmonary fibrosis: CT patterns and correlation with pulmonary function. AB - OBJECTIVE: The purpose of our study was to identify CT patterns of pulmonary fibrosis in patients with sarcoidosis and to correlate these patterns with pulmonary function tests. MATERIALS AND METHODS: We conducted a retrospective review of CT scans of 80 patients with proven sarcoidosis and evidence of fibrotic changes on chest radiographs. RESULTS: Three main CT patterns were identified: bronchial distortion (n = 38, 47%), mainly central; honeycombing (n = 23, 29%), mainly peripheral; and linear (n = 19, 24%), mainly diffuse. In most cases, a pattern was clearly identified as shown by the good agreement between observers (kappa = 0.87). Nodules were significantly associated with the linear (87%) and distorted (71%) patterns, but not with the honeycomb pattern (35%). The honeycomb pattern was most often associated with restriction and decreased lung diffusing capacity for carbon monoxide. Patients with bronchial distortion had lower expiratory airflow rates. The linear pattern was generally associated with the least functional impairment. CONCLUSION: CT may be a useful tool for defining subgroups of patients with fibrotic pulmonary sarcoidosis. CT reveals three main patterns that may reflect different distributions of fibrotic lesions in the lung with different functional pulmonary impairments. The persistence of active pulmonary lesions suggested by the presence of nodular lesions was often associated with linear and distorted patterns. PMID- 10845519 TI - CT-guided transthoracic needle biopsy using an ipsilateral dependent position. AB - OBJECTIVE: We postulated that the pneumothorax rate of transthoracic needle biopsy might improve with an ipsilateral dependent position of the affected side. We tried to determine the feasibility, effectiveness, and safety of CT-guided biopsy with the patient in this position. SUBJECTS AND METHODS: CT-guided needle biopsy with the patient in an ipsilateral dependent position was performed in 23 patients with 17 lung lesions (15 posterior and two anterior) and six mediastinal lesions. Fine-needle aspiration was used in all patients, and core biopsy was also used in six patients. The technical difficulty of the procedure was classified into three grades compared with a routine transthoracic needle biopsy as follows: grade I, no more difficult; grade II, somewhat more difficult; and grade III, much more difficult. RESULTS: Adequate samples were obtained in 22 (96%) of 23 patients. A small asymptomatic pneumothorax occurred in two patients (8.7%). Difficulty was rated grades I, II, and III in 18 (78%), two (9%), and three (13%) procedures, respectively. Four of the five grades II and III procedures were biopsies of anterior lesions. Traversing the pleura was avoided in three of six mediastinal masses. CONCLUSION: Transthoracic needle biopsy of selected lung and mediastinal lesions using an ipsilateral dependent position is feasible, effective, and safe. The role of this technique for reducing the rate of pneumothorax as a result of the biopsy requires further investigation. PMID- 10845520 TI - Usefulness of the double-wall sign in detecting pneumothorax in patients with giant bullous emphysema. AB - OBJECTIVE: We describe a new sign improving detection of pneumothorax in patients with giant bullous emphysema: air surrounding both sides of the bulla wall (the intrathoracic equivalent of the double-wall sign of pneumoperitoneum). We report the radiographic and CT appearances of the double-wall sign in seven patients with giant bullous emphysema, four of whom had pneumothorax. CONCLUSION: Recognizing the double-wall sign of pneumothorax should aid in the triage of patients with giant bullous emphysema. PMID- 10845521 TI - Breast Imaging Reporting and Data System: inter- and intraobserver variability in feature analysis and final assessment. AB - OBJECTIVE: We sought to evaluate the use of the Breast Imaging Reporting and Data System (BI-RADS) standardized mammography lexicon among and within observers and to distinguish variability in feature analysis from variability in lesion management. MATERIALS AND METHODS: Five experienced mammographers, not specifically trained in BI-RADS, used the lexicon to describe and assess 103 screening mammograms, including 30 (29%) showing cancer, and a subset of 86 mammograms with diagnostic evaluation, including 23 (27%) showing cancer. A subset of 13 screening mammograms (two with malignant findings, 11 with diagnostic evaluation) were rereviewed by each observer 2 months later. Kappa statistics were calculated as measures of agreement beyond chance. RESULTS: After diagnostic evaluation, the interobserver kappa values for describing features were as follows: breast density, 0.43; lesion type, 0.75; mass borders, 0.40; special cases, 0.56; mass density, 0.40; mass shape, 0.28; microcalcification morphology, 0.36; and microcalcification distribution, 0.47. Lesion management was highly variable, with a kappa value for final assessment of 0.37. When we grouped assessments recommending immediate additional evaluation and biopsy (BI RADS categories 0, 4, and 5 combined) versus follow-up (categories 1, 2, and 3 combined), five observers agreed on management for only 47 (55%) of 86 lesions. Intraobserver agreement on management (additional evaluation or biopsy versus follow-up) was seen in 47 (85%) of 55 interpretations, with a kappa value of 0.35 1.0 (mean, 0.60) for final assessment. CONCLUSION: Inter- and intraobserver variability in mammographic interpretation is substantial for both feature analysis and management. Continued development of methods to improve standardization in mammographic interpretation is needed. PMID- 10845522 TI - Comparative value of 99mTc-sestamibi scintimammography and sonography in the diagnostic workup of breast masses. AB - OBJECTIVE: This study was conducted to assess the relative roles of 99mTc sestamibi scintimammography and sonography in the evaluation of breast lesions that are indeterminate or suspicious on mammography or clinical examination. SUBJECTS AND METHODS: Twenty-five patients with 33 biopsy-proven breast lesions underwent both scintimammography and sonography. Lesions were categorized as benign or requiring biopsy on the basis of the absence or presence of a focus of increased activity on scintimammography and the shape, orientation, and echogenicity of the lesion on sonography. RESULTS: Sensitivity and specificity in detecting breast cancer were 92% and 95%, respectively, for scintimammography and 100% and 48%, respectively, for sonography. The higher specificity of scintimammography was statistically significant (p < 0.01). CONCLUSION: Although the overall accuracy of 99mTc-sestamibi scintimammography in the diagnosis of breast cancer was high, it has several disadvantages in comparison with sonography. Scintimammography has a slightly higher false-negative rate for breast cancer, is unable to reveal cysts, is more expensive, takes longer to perform, and involves ionizing radiation. For these reasons, scintimammography with 99mTc-sestamibi is unlikely to either replace sonography or be frequently used in addition to sonography. PMID- 10845523 TI - Imaging of primary actinomycosis of the breast. PMID- 10845524 TI - Rethinking the argument against glucagon for CT colonography. PMID- 10845525 TI - Physiologic changes of the cross-sectional measurements of the spinal canal and intervertebral foramina as a function of body position. PMID- 10845526 TI - Stop compressing, start injecting. PMID- 10845527 TI - Duplicate publication or not? Case 1. PMID- 10845528 TI - Duplicate publication or not? Case 2. PMID- 10845529 TI - MR imaging findings of intramedullary lipomas. PMID- 10845530 TI - Diastematomyelia and epidermoid cyst in the hemicord. PMID- 10845531 TI - Patellar metastasis from a squamous cell carcinoma of the larynx. PMID- 10845532 TI - Tattoo-induced skin burn during MR imaging. PMID- 10845533 TI - Theoretical study of the force field and vibrational assignments of thioacetamide and its deuterated analogues. AB - The harmonic force field of thioacetamide has been obtained by Hartree-Fock level ab initio calculations using the 4-31G(d) basis set. The force constants have been scaled to reproduce the infrared fundamental bands of argon matrix isolated CH3CSNH2 and CD3CSNH2. Normal coordinate calculations based on the scaled force field suggested the co-presence of cis-CH3CSNHD and trans-CH3CSNHD in the infrared spectrum measured for CH3CSND2. The prediction of the fundamental bands was made for CD3CSND2, cis-CD3CSNHD and trans-CD3CSNHD. PMID- 10845534 TI - Study of infrared and mass spectra of N-2-(3'-phthalide-yl)phthalazin-1-one. AB - The infrared and mass spectra of N-2-(3'-phthalide-yl)phthalazin-1-one were studied. The loss of one or two natural CO residues was observed from the title compound and its partial fragment ions. Using the MIKE spectrum of ion m/z 250, its most possible structure was confirmed. Two possible paths of the formation of the title compound were also proposed. PMID- 10845535 TI - Structural and vibrational characterisation of 3-amino-1-propanol a concerted SCF MO ab initio, Raman and infrared (matrix isolation and liquid phase) spectroscopy study. AB - Results obtained for the isolated and liquid 3-amino-1-propanol by a concerted molecular orbital and vibrational spectroscopic approach are reported. The relative energies and both structural and vibrational data of the different conformers of the studied compound were calculated using the extended 6-31G* basis set both at the HF-SCF and MP2 ab initio levels of theory and the theoretical results used to interpret Raman and infrared experimental data. In the gaseous phase and for the molecule isolated in an Argon matrix, monomeric 3 amino-1-propanol exists as a mixture of conformers, the first and second lowest energy forms corresponding to conformers which exhibit an intramolecular OH-N hydrogen bond (forms I and II). On the other hand, in the pure liquid, where intermolecular H-bonding occurs, the monomeric unit within the aggregates assumes a conformation similar to that of the third most stable form found for the isolated molecule situation (form III), which is characterised by having a weak intramolecular NH-O bond. The experimental data obtained for the pure liquid also reveals the presence of monomeric form I in this phase, a result that is in consonance with the strongly stabilizing OH-N intramolecular hydrogen bond that is present in this conformer. PMID- 10845537 TI - 1H and 13C NMR spectral studies of some 4H-3,1-benzoxazin-4-ones and their 2 acylaminobenzoic acid precursors. AB - The 1H and 13C NMR spectra of twelve 4H-3,1-benzoxazine-4-ones and of their acylaminobenzoic acid precursors are presented. Differentiation between these two series of compounds is best achieved through the characteristic J(CH) coupling interactions in the high frequency carbonyl region. Some 4H-pyrido[2,3 d][1,3]oxazin-4-ones have also been studied and some earlier literature assignments revised. PMID- 10845536 TI - Infrared and Raman spectra, conformational stability, barriers to internal rotation, normal-coordinate calculations and vibrational assignments for vinyl silyl bromide. AB - The infrared (3200-30 cm(-1) spectra of gaseous and solid, the Raman spectra (3200-30 cm(-1)) of the liquid and solid vinyl silyl bromide, CH2CHSiH2Br, have been recorded. Additionally, quantitative depolarization values have been obtained. Both the gauche and cis conformers have been identified in the fluid phases but only the gauche conformer remains in the solid. Variable temperature studies from 0 to -87 degrees C of the Raman spectrum of the liquid was carried out. From these data, the enthalpy difference has been determined to be 22 +/- 6 cm(-1) (0.26 +/- 0.08 kJ/mol), with the gauche conformer being the more stable form. The predictions from the ab initio calculations up to MP2/6-311 + + G(2d,2p) basis set favor the gauche as the more stable form. A complete vibrational assignment is proposed for both the gauche and cis conformers based on infrared band contours, relative intensities, depolarization values and group frequencies. The vibrational assignments are supported by normal coordinate calculations utilizing the force constants from ab initio MP2/6-31G(d) calculations and the potential energy terms for the conformer interconversion have been obtained from the same calculations. Complete equilibrium geometries have been determined for both rotamers by ab initio calculations employing a variety of basis sets up to 6-311 + + G(2d,2p) at levels of restricted Hartree Fock (RHF) and/or Moller-Plesset (MP) to second order. The results are discussed and compared to those obtained for some similar molecules. PMID- 10845538 TI - Experimental and theoretical vibrational study of silyl trifluoromethanesulfonate, CF3SO2OSiH3. AB - Infrared and Raman spectra were obtained for liquid silyl trifluoromethanesulfonate, a silylating agent of limited stability. The molecular geometry was optimized by means of density functional theory and Moller-Plesset second order perturbation theory methods, using different basis sets. The optimized structure presents a gauche conformation, similar to that adopted by methyl trifluoromethanesulfonate, which was determined experimentally a short time ago. The wavenumbers for the normal modes of vibration and the corresponding force constants were also calculated, facilitating the interpretation of the vibrational data. The harmonic force constants given by theory were scaled to reproduce adequately the experimental wavenumbers. PMID- 10845539 TI - Absolute band intensities in the nu19/nu23 (530 cm(-1)) and nu7 (777 cm(-1)) bands of acetone ((CH3)2CO) from 232 to 295 K. AB - Absolute band intensities of acetone ((CH3)2CO) in the nu19/nu23 and nu7 band systems near 530 and 777 cm(-1), respectively, were measured at temperatures of 232, 262 and 295 K, using a Fourier transform infrared (FTIR) spectrometer. No evident temperature dependence for the band intensities was observed. The dipole moments and the fundamental band intensities were derived in the harmonic oscillator approximation. The results are useful for the spectroscopic retrieval of acetone concentrations in the upper atmosphere. PMID- 10845540 TI - Photophysical properties of 5H-dibenzo(a,d)cyclohepten-5-one in different solutions. AB - A number of photophysical properties of 5H-dibenzo(a,d)cyclohepten-5-one have been measured for the first time including the prompt and P-type delayed fluorescence, and triplet-triplet absorption. Decay parameters, for the triplet state of this molecule, have also been measured in different quenching conditions. The results are discussed and compared for those of similar molecules. PMID- 10845541 TI - The vibrational spectrum of methyltrioxorhenium(VII). AB - The inelastic neutron scattering and high-resolution FT-Raman spectra of methyltrioxorhenium are reported. Literature assignments of the modes below 1600 cm(-1) are largely confirmed with the exceptions of the methyl torsion and the ReO3 bending modes. The methyl torsion is observed for the first time at 200 cm( 1), while the band at 230 cm(-1) assigned previously to the torsion is shown to be a component of the factor group splitting of the solid. In the FT-Raman spectrum, the bands that derive from the E modes in the isolated molecule are split by the Cs site symmetry of the crystal. This allows an unambiguous assignment of these modes to be made. On this basis, the assignment of the ReO3 bending modes is reversed with the asymmetric mode at 325 cm(-1) and the symmetric mode at 276 cm(-1). PMID- 10845542 TI - Theoretical calculation of the vibrational spectra of cis-cis-cyclooctadienes in the vapour phase. AB - The theoretical infrared spectra of 1,3-cis-cis-cyclooctadiene (1,3-COD) and 1,5 cis-cis-cyclooctadiene (1,5-COD), were obtained by ab initio MO calculations at Hartree-Fock level. The results were compared with the available IR experimental spectra of 1,3- and 1,5-COD. The apparent agreement between theoretical and experimental data allows us to exploit two bands, found only in the case of the theoretical spectrum of 1,4-COD, as a tool for identifying 1,4-COD during its synthesis. PMID- 10845543 TI - IR study of dickite-formamide intercalate, Al2Si2O5(OH)4-H2NCOH. AB - Direct intercalation of formamide (FAM) in dickite occurs spontaneously when samples are treated by ultrason. The X-ray diffraction patterns show that this intercalation increases the d001 spacing from 7.19 to 10.77 A. It is concluded from infrared studies that hydrogen bonds are formed between C=O groups of formamide and inner surface hydroxyls of dickite, indicated by the shift of the hydroxyl bands from 3708, 3654 cm(-1) and 3622 for natural dickite to 3575, 3520, 3450 and 3612 cm(-1) for FAM-intercalated dickite. PMID- 10845544 TI - Vibronic interaction in europium nitrates Eu(NO3)3 x 4SOR2. AB - Luminescence and excitation of luminescence vibronic spectra of europium nitrates Eu(NO3)3 x 4SOR2 containing sulphoxide derivatives were obtained and analysed. Some factors influencing the intensity distribution in vibronic sidebands are discussed. Significant variation of the intensity distribution in antiStokes sidebands of Eu3+ electronic transitions in series of nitrates results from the difference in effective charges on coordinated oxygen atoms of ligands. Another important detail of the vibronic spectra is a redistribution of intensity in the region of 5D0, 5D1-->7F2 transitions of luminescence spectra originated in overlap of different vibronic transitions. Mixing between the 7F2 electronic state of Eu3+ and vibronic satellites of 7F0 electronic state was studied both under conditions of resonance and in case of significant detuning. PMID- 10845545 TI - PM3, AM1, MINDO3 semi-empirical IR spectra simulations for some nitriles of interest for Titan's chemistry. AB - A set of the semi-empirical methods (PM3, AM1, MNDO and MINDO3) has been tested to find the best auxiliary tool for the identification of nitriles by gas chromatography/Fourier transform IR spectroscopy/mass spectrometry, considering five nitriles of interest for Titan's chemistry as test compounds: acetonitrile, acrylonitrile, cyanoacetylene, 2-butynenitrile and dicyanoacetylene. Of the four semi-empirical methods, MNDO can be considered as the most advantageous auxiliary tool for the gas chromatography/Fourier transform IR spectroscopy/mass spectrometry (GC/FTIR/MS) identification of nitriles of interest for Titan's atmospheric chemistry, since (1) the simulated IR spectra best match the experimental (in some cases AM1 gives comparable results); (2) it provides the best linearity between the calculated and experimental frequencies (correlation coefficient of 0.990); a scaling factor of 0.90 can be applied to afford better correspondence between the calculated and experimental wavenumbers. At the same time, none of the methods is able to predict infrared intensities and a spectral intensity pattern. PMID- 10845546 TI - Synthesis and experimental study of through-space hydrogen-fluorine and carbon fluorine spin-spin coupling in 4,5-substituted 1-acetyl-8-fluoronaphthalenes. AB - The synthesis and NMR study (1H, 13C, and 19F) of a complete series of 4,5 substituted 1-acetyl-8-fluoronaphthalenes are reported. This data revealed a 6J(H,F) and a 5J(C,F) through space coupling between the fluorine and the methyl on the acetyl group (1H and 13C). The magnitude of this coupling constant changes depending on the nature of the substituent at C-4, the internuclear distance, and the solvent. PMID- 10845547 TI - Structural chemistry of polycyclic heteroaromatic compounds. Part XI. Photoelectron spectra and electronic structures of tetracyclic hetarenes of the triphenylene type. AB - The UV photoelectron spectra of several tetracyclic heteroaromatic compounds (2 9) which are pi-isoelectronic with triphenylene (1) have been recorded and analysed making use of semiempirical AM1 and PM3 as well as ab initio/DFT B3LYP calculations. In one series of compounds (2-7), the peripheral benzene rings of 1 are successively substituted by thiophene rings that are either [b]- or [c] annellated with the central benzene unit. In 2-7 only marginal shifts are found for most of the IPs of electrons. In the benzotrithiophenes 5-7, a systematic variation is displayed by IP(pi7). Compared to 1, the pi electron system of benzo[c]trithiophene (7) is approximately two times as much destabilized as in the isomers 5 and 6 with [b]annellated thiophene rings. The IP[n(S)] values of the thiophene derivatives 2-7 indicate that these orbitals are clearly destabilized relative to thiophene. The same holds for the n(O) orbital of the furane derivative 9 in comparison with that of furane. In 9, only the higher pi MOs (pi7-pi9) are destabilized whereas the lower levels (pi1-pi4) are stabilized, and those in between (pi5-pi6) remain essentially unshifted. In the pyrrole derivative 8, all pi MOs are substantially destabilized by about 0.5-1.6 eV relative to 1. PMID- 10845548 TI - Vibrational spectroscopy of formamide-intercalated kaolinites. AB - The vibrational spectroscopy of low and high defect kaolinites fully and partially intercalated with formamide have been determined using a combination of X-ray diffraction, DRIFT and Raman spectroscopy. Expansion of the high defect kaolinite to 10.09 A resulted in a decrease in the peak width of the d(001) peak attributed to a decrease in defect structures upon intercalation. Changes in the defect structures of the low defect kaolinite were observed. Additional infrared bands were observed for the formamide intercalated kaolinites at 3629 and 3606 cm(-1). The 3629 cm(-1) band is attributed to the hydroxyl stretching frequency of the inner surface hydroxyl group hydrogen bonded to the carboxyl group of the formamide. The 3606 cm(-1) band is ascribed to water in the interlayer. Concomitant changes are observed in both the hydroxyl deformation modes and in the carboxyl bands. PMID- 10845549 TI - Photophysics of TICT states of 7-diethylamino-4-methyl coumarin dye by energy transfer techniques. AB - Coumarin 1 exhibited dual amplified spontaneous emission (ASE) in certain solvents under nitrogen laser excitation. These emissions are known as normal and anomalous emissions. The anomalous emission corresponds to TICT state and it does not have a corresponding fluorescence peak. Energy transfer techniques have been used to study the photophysics of TICT states and the characteristics of dual ASE bands of the dye coumarin 1. PMID- 10845550 TI - Luminescence behaviour of 5-hydroxyindole in different environments. AB - Steady state fluorescence emission spectroscopic studies along with some lifetime measurements have been performed for 5-hydroxyindole (5HI) in different environments. 5HI merits particular attention, since it is the chromophoric moiety of the non-natural amino acid 5-hydroxytryptophan (5HT), which has come into significant, recent prominence as a novel intrinsic optical probe for protein structure, function and dynamics. Studies in representative homogeneous solvents and solvent-mixtures indicate that unlike other fluorophores of related interest like indole (I) and 7-azaindole (7AI), the fluorescence emission maximum (lambda(em)max) of 5HI is relatively insensitive to solvent polarity. This behaviour suggests the lack of appreciable solvent dipolar relaxation in 5HI, which is consistent with our low temperature (77 K) emission data. Notwithstanding such limitation, fluorescence anisotropy (r) and quenching studies are shown to be effective for exploring changes in the micro-environments of 5HI in sodium bis-(2-ethylhexyl)sulfosuccinate (AOT) reverse micellar assemblies (which serve as a biomembrane mimetic model system) with variation in water/surfactant molar ratio (w0). PMID- 10845551 TI - Fine-structure enhancement--assessment of a simple method to resolve overlapping bands in spectra. AB - A simple mathematical procedure--fine-structure enhancement--has been assessed on its ability to resolve overlapping bands in spectra. Its advantages and limitations have been explored using synthetic and experimental spectra. Fine structure enhancement involves smoothing the original spectrum, multiplying the smoothed spectrum with a weighting factor and subtracting this spectrum from the original spectrum. As a result, the fine-structure of the original spectrum is enhanced in the processed spectrum and bands that overlap in the original spectrum appear as distinct bands in the processed spectrum. To be resolved by fine-structure enhancement, Lorentzian lines have to be separated by more than their quarter width at half maximum, Gaussian lines by more than their half width at half maximum. A comparison of fine-structure enhancement and Fourier self deconvolution shows that Fourier self-deconvolution has in theory a higher potential to resolve overlapping bands. However, this depends crucially on the correct choice of the parameters. In practice, when parameters commonly used are chosen for Fourier self-deconvolution, fine-structure enhancement leads to similar results. This is demonstrated at the example of the infrared absorbance spectrum of the protein papain, where the amide I band components could be resolved similarly with both methods. Thus, fine-structure enhancement seems to be a simple alternative to Fourier self-deconvolution that does not require specialised software. PMID- 10845552 TI - Interaction of three pradimicin derivatives with divalent cations in aqueous solution. AB - This study focused on further analysis of the aggregation behavior of pradimicin derivatives and their interaction with cations in aqueous solution. BMY was compared with two other pradimicin antibiotics (T2 and FB) with the same aglycone moiety but consisting of different substitute groups. The surface tension measurement showed a clear critical micelle concentration at 1-2 mM of the BMY aqueous solution. The role of Zn2+ in replacing the Ca2+ was examined using 1H nuclear magnetic resonance (NMR) method. From changes in the NMR spectrum and precipitability, it was concluded that zinc ion has lower affinity and higher precipitating ability to BMY than the divalent cations of alkaline earth metal. The aggregation behavior of T2 and FB in aqueous solution was also studied using NMR method. The results suggest that the supramolecular behavior of T2 is similar to BMY whether or not Ca2+ ions are present in solution and that there are two binding sites for calcium ions in a T2 molecule. Unlike BMY and T2, the NMR spectrum of FB does not show distinct change upon Ca2+ addition. The interaction of pradimicin antibiotics with divalent metal ions was thought to be related to ionic electronegativity and to the amphoteric property of the antibiotics. PMID- 10845553 TI - bcl-2 and c-erbB-2 proteins are involved in the regulation of VEGF and of thymidine phosphorylase angiogenic activity in non-small-cell lung cancer. AB - Tumour angiogenesis has been recently recognised as one of the most important prognostic factors in lung cancer. Although a variety of angiogenic factors have been identified, the angiogenesis process remains poorly understood. Bcl-2, c erbB-2 and p53 are well-known oncogenes involved in non-small-cell lung cancer pathogenesis. A direct correlation of thymidine phosphorylase (TP) and of vascular endothelial growth factor (VEGF) with intratumoural angiogenesis has been reported. In the present study we investigated the possible regulatory role of bcl-2, c-erB-2 proteins in angiogenesis and in VEGF and TP expression in non small-cell lung cancer. Two hundred sixteen specimens from T1,2-N0,1 staged patients treated with surgery alone were immunohistochemically examined. Bcl-2 and c-erbB-2 were significantly inversely related to each other (P = 0.04) and both were inversely associated with microvessel density (P < 0.02). High TP and VEGF reactivity was statistically related to loss of bcl-2 expression (P < 0.01). A significant co-expression of c-erbB-2 with TP was noted (P = 0.01). However, TP expression was related to high angiogenesis only in cases with absence of c-erB-2 expression (P < 0.0001). c-erbB-2 expression in poorly vascularised tumours was linked with poor outcome (P = 0.03). The present study provides strong evidence that the bcl-2 gene has a suppressive function over genes involved in both angiogenesis (VEGF and TP) and cell migration (c-erbB-2) in NSCLC. TP and c-erbB 2 proteins are significantly, and often simultaneously, expressed in bcl-2 negative cases. However, expression of the c-erbB-2 abolishes the TP-related angiogenic activity. Whether this is a result of a direct activity of the c-erbB 2 protein or a consequence of a c-erbB-2-related immune response remains to be further investigated. PMID- 10845554 TI - Expression of matrix metalloproteinases and their inhibitors in human brain tumors. AB - Sixty human brain tumors, classified according to the New World Health Organization (WHO) classification including, grade I schwannomas, meningiomas and pilocytic astrocytomas, grade II astrocytomas, grade III anaplastic astrocytomas, grade IV glioblastomas, grade III anaplastic oligodendrogliomas and grade IV glioblastomas and lung and melanoma metastases were analyzed for the expression of three matrix metalloproteinases (MMPs), two tissue inhibitors of MMPs (TIMPs) and for MMP activity. Some correlation was found between MMP expression and the degree of malignancy. Western blotting analysis revealed a more uniform pattern of distribution of MMP-2 (gelatinase A) than of MMP-9 (gelatinase B) and MMP-12 (metalloelastase) among tumors. MMP-9 levels were found to be significantly higher in grade III anaplastic astrocytomas and anaplastic oligodendrogliomas than those in grade I schwannomas and meningiomas. Anaplastic astrocytomas and Grade IV glioblastomas expressed significantly higher levels MMP-12 than grade I meningiomas. All sixty tumors showed a similar pattern of activity in zymography, proMMP-9 being the major species detected. Interestingly, TIMP-1 and TIMP-2 expression levels were especially low in tumors of grade II and grade III but significantly higher in tumors of grade I, particularly in schwannomas. Taken together, these data suggest that: 1) a balance between MMPs and TIMPs has an important role to play in human brain tumors; 2) TIMP expression may be valuable markers for tumor malignancy. PMID- 10845555 TI - Growth-associated changes in glutathione content correlate with liver metastatic activity of B16 melanoma cells. AB - B16 melanoma (B16M) was used to study the relationship between glutathione (GSH) metabolism and the metastatic activity of malignant cells. GSH content increased in B16M cells during the initial period of exponential growth in vitro, to reach a maximum of 37 +/- 3 nmol/10(6) cells 12 h after plating, and then gradually decreased to control values (10 +/- 2 nmol/10(6) cells) when cultures approached confluency. On the contrary, glutathione disulphide (GSSG) levels (0.5 +/- 0.2 nmol/10(6) cells) and the rate of glutathione efflux (GSH + GSSG) (2.5 +/- 0.4 nmol/10(6) cells per h) remained constant as B16M grew. Changes in enzyme activities involved in GSH synthesis or the glutathione redox cycle did not explain shifts in the glutathione status (GSH/GSSG). However, two facts contributed to explain why GSH levels changed within B16M cells: a) high intracellular levels of GSH induced a feed-back inhibition of its own synthesis in B16M cells from cultures with low cellular density (LD cells); b) transport of cyst(e)ine, whose availability is the major rate-limiting step for GSH synthesis, was limited by cell-cell contact in cultures with high cellular density (HD cells). Intrasplenic injection of B16M cells with high GSH content (exponentially growing cultures) showed higher metastatic activity in the liver than cells with low GSH content (cells at confluency). However, when low GSH-content cells (HD cells) were incubated in the presence of GSH ester, which rapidly enters the cell and delivers free GSH, their metastatic activity significantly increased. Our results demonstrate that changes in GSH content regulate the metastatic behaviour of B16M cells. PMID- 10845556 TI - Antiangiogenic, antitumoural and antimetastatic effects of two distamycin A derivatives with anti-HIV-1 Tat activity in a Kaposi's sarcoma-like murine model. AB - The antiangiogenic, antitumoural and antimetastatic effects of two novel sulphonic derivatives of distamycin A, PNU145156E and PNU153429, were studied in a Kaposi's sarcoma-like tumour model obtained by injecting nude mice with cells releasing extracellular HIV-Tat protein, derived from a tumour which developed in a BK virus/tat transgenic mouse. Both PNU145156E and PNU153429 were administered intraperitoneally every fourth day for three weeks at doses of 100 or 50 mg/kg of body weight respectively, starting one day after injecting the tumour cells. Both drugs delayed tumour growth in nude mice, preventing neovascularization induced by the Tat protein. PNU153429 also significantly reduced the number and size of spontaneous tumour metastases. Both effects on tumour growth and metastases were augmented by treating simultaneously nude mice with 7.5 mg/kg of body weight of minocycline given per os daily for four weeks starting four days after injecting the tumour cells. Neither acute nor chronic toxic side-effects were observed during the life span of treated nude mice. Due to their antiangiogenic and anti Tat effects, these drugs are promising for the treatment of Kaposi's sarcoma in AIDS patients. PMID- 10845557 TI - Membrane protein glycosylation and CD44 content in the adhesion of human ovarian cancer cells to hyaluronan. AB - The adhesion of tumour cells to the hyaluronan (HA) pericellular coat of mesothelial cells is an important step in the peritoneal spread of ovarian cancer. Previously, we have shown that the cell surface molecule CD44 is involved in this process. Paradoxically, the degree of adhesion does not appear to be related to the amount of CD44 expressed. In order to explain this observation we have examined the in vitro adhesion to HA of four high CD44-expressing ovarian cancer lines in relation to their CD44 spliced variant content and the CD44 glycosylation. Adhesion was measured in multiwell plates coated with different concentrations of HA in order to determine both the avidity and the maximum adhesion. Two lines had high adhesion and two lines had low adhesion. The avidity for HA was different for each line, but in all cases this could be totally blocked by treatment with an anti-CD44 antibody. The standard form of CD44 was the major species detected by RT/PCR in all lines and spliced variants were present in low amounts. Neuraminidase treatment increased the adhesion of the 'low-adhesion' lines at all HA coating concentrations; but only substantially increased the adhesion of the 'high-adhesion' lines at the lower HA coating concentrations. Tunicamycin treatment decreased the adhesion of the 'high adhesion lines' at all HA coating concentrations and only substantially decreased the adhesion of one of the 'low-adhesion' lines when the plates were coated with a low concentration of HA. The adhesion of the remaining 'low-adhesion' line was slightly increased after tunicamycin treatment. It is concluded that glycosylation and not spliced variant content of CD44 affects the adhesive properties of ovarian tumour cells. This conclusion may have important consequences for developing new therapies in ovarian cancer. PMID- 10845558 TI - Genetic diagnosis of micrometastasis based on SCC antigen mRNA in cervical lymph nodes of head and neck cancer. AB - This study is designed to assess gene expression of squamous cell carcinoma antigen (SCCA) mRNA to detect micrometastases in cervical lymph nodes (LNs) of head and neck cancer. We examined the expression of SCCA mRNA in 12 primary tumors and 212 cervical LNs (101 LNs taken from 8 patients with tongue cancer, 71 from 7 patients with gingival cancer, 19 from 2 patients with laryngeal cancer, 9 from 2 patients with pharyngeal cancer, 7 from 1 patient with cancer of the buccal mucosa, and 5 from 1 patient with cancer of floor of the mouth). Detectability of metastatic LNs by nested and single reverse transcriptase polymerase chain reaction (RT-PCR) was compared with semiserial sections (hematoxylin-eosin staining and keratin immunostaining). All primary tumors expressed SCCA mRNA. Of 198 histologically metastasis-negative nodes, SCCA mRNA was detected in 37 (18.7%) by nested PCR. Eleven micrometastatic foci in 9 LNs (4.6%) were discovered by semiserial sectioning. This suggests that SCCA mRNA is a promising tumor marker for detecting the micrometastases in cervical LNs of head and neck cancer. PMID- 10845559 TI - Metastasis-suppressed C8161 melanoma cells arrest in lung but fail to proliferate. AB - The incidence of melanoma continues to increase at a rapid rate. As for most cancers, it is melanoma metastases, rather than the primary malignancy, that is the principal cause of death. We previously showed that the introduction of a normal copy of chromosome 6 into the metastatic human melanoma cell line C8161 suppresses metastasis at a step subsequent to tumor cells entering the bloodstream. To better define the step(s) in metastasis blocked by the addition of chromosome 6 we engineered cells that constitutively express green fluorescent protein (GFP). When these tagged, chromosome 6 hybrid cells were injected intravenously into athymic mice, grossly detectable metastases did not form. However, fluorescence microscopy revealed micro-metastases (single cells or clusters of <10 cells) in the lungs, suggesting that these cells lodged in the lungs but failed to proliferate. Cells isolated from lung up to 60 days post injection grew in culture and/or formed tumors when injected into the skin, indicating that they were still viable, but dormant. This result implies that the gene(s) on chromosome 6 interfere specifically with growth regulatory response in the lung, but not in the skin. Thus, the gene(s) responsible for metastasis suppression represents a new class of metastasis inhibitors acting at the final stages of the metastatic cascade--that is, affecting the ability of the cells to survive and proliferate at a specific secondary site. PMID- 10845560 TI - Is the Fischer 344/CRJ rat a protein-knock-out model for dipeptidyl peptidase IV mediated lung metastasis of breast cancer? AB - Fischer 344/CRJ rats harbor a G633R substitution in dipeptidyl peptidase IV (DPP IV) that leads to retention and degradation of the mutant protein in the endoplasmic reticulum (Tsuji E, Misumi Y, Fujiwara T et al. Biochemistry 1992; 31 (47): 11921-7). However, when these rats were used as a 'protein knock-out' model in further evaluating the previously established role of DPP IV in metastasis, lung colonization of the highly metastatic MTF7 rat breast cancer cell line was reduced by only 33% relative to normal Fischer 344 rats. To examine whether lung endothelia leak expression of mutant DPP IV and whether mutant DPP IV exhibits the same adhesion qualities as wild type DPP IV, detailed immunohistochemical, biochemical, transfection, and FACS analyses were performed to assess the surface expression of mutant DPP IV on lung endothelia and transfected HEK293 cells and adhesion assay to compare the adhesion qualities of wild-type and mutant DPP IV. Both endothelial and transfected HEK293 cells expressed mutant, enzymatically inactive DPP IV on their surfaces, albeit at greatly reduced levels when compared to expression of wild type DPP IV. Purified mutant DPP IV had identical adhesion qualities for lung-metastatic MTF7 cells as wild type DPP IV, and competitive inhibition of MTF7 lung colonization by truncated DPP IV confirmed involvement of mutant DPP IV in lung metastasis of Fischer 344/CRJ rats. Although metastasis appears to be mediated by several, often parallel mechanisms involving multiple tumor and host factors, these data indicate that altered expression of a single component can drastically change the outcome of metastatic disease. PMID- 10845561 TI - Downregulation of the urokinase-type plasminogen activator receptor through inhibition of translation by antisense oligonucleotide suppresses invasion of human glioblastoma cells. AB - We previously showed that downregulation of the urokinase-type plasminogen activator receptor (uPAR) in the SNB19 human glioblastoma cell line by the stable transfection of a plasmid expressing a 300 bp antisense sequence to the 5' end of the uPAR gene produced a decrease in the amount of target mRNA. In a more recent study, we found that adenovirus-mediated transduction (Ad-uPAR) of the same uPAR antisense gene construct in SNB19 cells also downregulated uPAR protein levels. We report here that Ad-uPAR-transfected SNB19 cells produced the same amounts of target uPAR mRNA but significantly less protein by in vitro translation and by in situ [35S] labeling compared to Ad-CMV vector-transfected and mock-transfected cells. This antisense construct also inhibited glioblastoma cell invasion confirming previous results. We conclude that downregulation of uPAR by this antisense gene construct results from inhibition of protein translation. PMID- 10845562 TI - Significance of VLA-4-VCAM-1 interaction and CD44 for transendothelial invasion in a bone marrow metastatic myeloma model. AB - In previous work, we established the B9/BM1 syngeneic murine bone marrow metastasis model. Interleukin (IL)-6-dependent. IL-1-producing B9/BM1 cells, which colonize the vertebral and femoral marrow after i.v. injection, show great similarity in cell surface phenotype to human myeloma cells, especially the expression of 3 adhesion molecules, CD44, VLA-4 and ICAM-1. Here we investigated the function of these adhesion molecules by binding and transendothelial invasion assays using a newly established bone marrow-derived endothelial cell line (BMEC). A combination of monoclonal antibodies against CD44 and VLA-4 significantly inhibited the adherence of B9/BM1 cells to BMEC and anti-CD44 mAb especially blocked B9/BM1 transendothelial invasion of unstimulated BMEC cells. Results of additional experiments, in which the cells were treated with anti-CD44 and hyaluronidase, demonstrated that the interaction of CD44 molecules on B9/BM1 cells with hyaluronan on BMEC cells was a critical factor in both adhesion and transendothelial invasion in this model. However, stimulation of BMEC with TNFalpha resulted in increased invasion by B9/BM1 cells, which was completely suppressed by anti-VCAM-1 mAb, implicating a significant role of this adhesion molecule in this process during inflammation. PMID- 10845564 TI - Mouse models for human disease. PMID- 10845563 TI - Differential modulation of proliferation, matrix metalloproteinase expression and invasion of human head and neck squamous carcinoma cells by c-erbB ligands. AB - Evidence suggests that there is an association between the abnormal expression of members of the c-erbB receptor tyrosine kinase family and poor prognosis in head and neck squamous cell carcinomas (HNSCC). Until now, the relative contributions of different c-erbB ligands to HNSCC progression have not been clearly defined. In this paper we examined the effects of ligands with different c-erbB receptor specificities in terms of their stimulation of HNSCC proliferation, expression of matrix metalloproteinases (MMPs) and invasion. Heregulin-beta1 (HRG-beta1; selective c-erbB3/B4 ligand) was found to stimulate proliferation in the majority of cell lines, whereas epidermal growth factor (EGF; EGFR ligand) and betacellulin (BTC; EGFR/B4 ligand) induced variable responses. All three ligands up-regulated multiple MMPs including collagenases, stromelysins, matrilysin and gelatinase B (MMP-9) but had minimal or no effects on gelatinase A (MMP-2), MT1 MMP and tissue inhibitors of MMPs (TIMPs). MMP-9 mRNA was induced to a higher level than other MMPs, although with slower kinetics. HRG-beta1 was less active than EGF and BTC at the optimal concentration (relative potency of EGF:BTC:HRG = 3:4:1). In vitro invasion through Matrigel was also increased by all three ligands in proportion to their MMP up-regulation. A specific anti-EGFR monoclonal antibody (mAb ICR62) inhibited MMP up-regulation, migration and invasion induced by all three ligands, whereas an anti-c-erbB-2 mAb ICR12 inhibited mitogenic and motogenic responses following ligand stimulation but had no effect on MMP expression. These results suggest that c-erbB ligands may differentially potentiate the invasive phenotype of HNSCC via co-operative induction of cell proliferation, migration and proteolysis. The EGFR signalling pathway appears to be the dominant component controlling the proteolytic and invasive phenotype in HNSCC, whereas the c-erbB-2 signalling pathway is responsible, in part, for the mitogenic and motogenic effects of ligands. PMID- 10845565 TI - Online Mendelian Inheritance in Man (OMIM) as a knowledgebase for human developmental disorders. AB - In this review, we summarize the current genetic information on human developmental disorders found in Online Mendelian Inheritance in Man (OMIM). The OMIM catalogues human phenotypes and genotypes and relevant mouse models. Among the more than 11005 genetic disorders and loci, we found at least 1231 human conditions with known gene mutationsffWe searched for human developmental disorders that present with structural defects during the perinatal period, and identified 162 such entries. We classified these entries by phenotypic features (e.g., skeletal dysplasias, axis and laterality defects, or eye disorders) and by the type of gene mutated (e.g., genes coding for transcription factors, structural proteins, enzymes, or receptors). Thirty-eight entries have allelic variants with gene mutations causing different functional consequences, thereby altering their interactions with modifying genes. Thirty-two entries show genetic heterogeneity due to either functional redundancy of more than one gene or genes that interact in common developmental pathways. Although many different types of genes are mutated in developmental disorders, we found that the disease genes are transcription factors in 49 entries. Mouse models are available for many of the human conditions, with the majority of these mutants being secondary to null mutations. These data allow us to begin to elucidate the complex developmental pathways involved in the molecular pathogenesis of human malformations. PMID- 10845566 TI - Genetic landmarks through philately--Karl Landsteiner: the father of blood grouping. PMID- 10845567 TI - Gender-specific effects of NAT2 and GSTM1 in bladder cancer. AB - One approach for risk assessment of cancer is the evaluation of polymorphic enzymes involved in cancer using molecular tools. Phase II enzymes are involved in the detoxification of several drugs, environmental substances and carcinogenic compounds. Here, we analyzed enzymes for their putative relevance in urinary bladder cancer. The hereditable enzyme polymorphism of arylamine N acetyltransferase 2 (NAT2) and glutathione S-transferase M1 (GSTM1) and T1 (GSTT1) was studied in 157 hospital-based patients and in 223 control subjects. Slow acetylation was not observed to be a significant risk factor of developing bladder cancer (OR: 1.33; 95% CI 0.85-2.09). One genotype responsible for slow acetylation (NAT2*5B/*6A) was observed significantly more frequently in bladder cancer patients compared with control subjects (OR: 1.63; 95% CI 1.03-2.58). Gender-specific effects were observed when patients were divided into subgroups. In male patients, slow acetylators were identified as carrying a significant increased risk of developing bladder cancer, in particular when the genotype NAT2*5B/*6A was combined with the GSTM1 null genotype (OR: 4.39; 95% CI 1.98 9.74). By contrast, the same genotype combination significantly protected female patients from bladder cancer (OR: 0.21; 95% CI 0.06-0.80). PMID- 10845568 TI - Localization of an acromesomelic dysplasia on chromosome 9 by homozygosity mapping. AB - The acromesomelic dysplasias (AMDs) are a group of genetic disorders that primarily affect the middle and distal segments of the extremities. A form of AMD is present on the isolated island of St Helena in the South Atlantic, which has a population of approximately 5500 derived from a number of founder individuals. DNA from four affected individuals and 11 first-degree relatives in four related nuclear families segregating an AMD was collected for gene mapping studies. Six consecutive markers on chromosome 9, spanning an approximately 5 cM region, showed identical homozygosity in all affected individuals, thus identifying a region of homozygosity by descent. Multipoint analysis generated a maximum lod score of Z = 2.85. These data localize the gene for this dysplasia to the pericentromeric region of chromosome 9 where the gene for the Maroteaux form of AMD is situated. The identification of the gene responsible for this disorder may shed further light on the complex processes involved in limb morphogenesis. PMID- 10845569 TI - Mutation and transcription analysis of transthyretin gene in Italian families with hereditary amyloidosis: a putative novel hot spot' in codon 47. AB - Transthyretin gene mutations are associated with autosomal dominant familial amyloidosis. The commonest phenotype in the patients is peripheral neuropathy, but restrictive cardiomyopathy is also a frequent sign. More than 70 different mutations in the gene have been described. Although these mutations are randomly distributed, some hot spots have also been reported notably at position 6, 30, 33, 58, 109, 119 and 122. A few of these codons contain a CpG dinucleotide. We describe an additional 'hot spot' occurring at codon 47, in which we report one novel and two previously described mutations. This codon, however, does not contain a CpG dinucleotide, suggesting that other mechanisms might be responsible for the allelic heterogeneity. All the reported mutations in codon 47 are located in the exon 2 consensus sequence and are potentially involved in splicing. We performed transcription analysis on two livers obtained from transplanted patients carrying the Ala47 mutation. These livers showed a normally spliced message, indicating that this mutation does not affect splicing. PMID- 10845570 TI - Molecular analysis of Y chromosome long arm structural instability in patients with gonadal dysfunction. AB - We have performed cytogenetic and molecular analyses of 45,X mosaics involving structurally abnormal Y chromosomes. Karyotypes were performed by standard cytogenetic methods and, in some cases, by fluorescence in situ hybridization, to distinguish monocentric and dicentric chromosomes. In addition, the deletions of Yq have been mapped using Southern blotting and polymerase chain reaction analysis. This paper provides additional information on the analysis of Y chromosome aberrations, and suggests that the stability of the Y chromosome in these instances is related to the site of the break point on Yq. PMID- 10845571 TI - Association of the R485K polymorphism of the factor V gene with poor response to activated protein C and increased risk of coronary artery disease in the Chinese population. AB - Inherited predisposition to thrombosis contributes to the initiation and progression of coronary artery disease (CAD). The present study was designed to explore the relationship between genetic variation of coagulation factor V and occurrence of CAD. A total of 141 unrelated patients with CAD and 175 healthy controls were analyzed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) for variation detection in all 25 exons of the factor V gene. Among the study subjects, 55 CAD patients and 73 controls were evaluated at random for response to activated protein C (APC) by Coatest APC resistance test. Polymorphisms in exon 4, 10, 13 and 16 of factor V gene were documented [642G-->T(S156), 1628--> A(R485K), 4070A-->G(H1299R) and 5380G A(V1736M), respectively]. The study also identified a novel polymorphism 327A G in exon 2 which did not alter the amino acid residue. Leiden mutation (R506Q) was not detected in any of our 316 subjects. Among the five polymorphisms, the allele frequency of 1628G--> A was significantly different between the CAD patients and the controls (0.36 vs. 0.21, p < 0.05). Subjects homozygous or heterozygous for the A allele of 1628G-->A polymorphism had lower normalized APC ratios than those with the GG genotype in the CAD group (1.16+/-0.13 and 1.18+/-0.23 vs. 1.36+/ 0.33, p <0.05) and in the controls, indicating that A(1628) allele was associated with a poor response to APC. We conclude that the 1628G-->A (R485K) polymorphism of factor V is associated with a poor response to APC and increased risk for CAD. PMID- 10845572 TI - Apolipoprotein B-100 gene Xba I polymorphism and cholesterol gallstone disease. AB - The apolipoprotein (apo) B gene Xba I polymorphism is associated with alterations in serum lipids. Disturbances in serum lipids may be a risk factor for cholesterol gallstone disease. However, the relation between the Xba I polymorphism and cholesterol gallstones is unknown. This study was aimed at characterizing the polymorphism of the apo B gene Xba I in patients with gallbladder stones and the association of Xba I polymorphism with serum lipids. Xba I genotypes were measured by PCR-RFLP, and serum lipids assayed in 190 patients with gallbladder stones and 441 control subjects. The frequency of the X+/- genotype (20.63 vs. 7.94%) and X+ allele (10.79 vs. 3.97%) was significantly higher in the patient group than in the control group. Patients with the X+/- genotype had a significantly higher concentration of total cholesterol, low density lipoprotein (LDL)-cholesterol, and apo B in serum than patients with the X-/- genotype. The X+ allele of the apo B gene is characterized by a higher cholesterol concentration and a higher LDL-cholesterol concentration in serum, and it may be a marker for increased risk of cholesterol gallstone disease. PMID- 10845573 TI - Relation between the insertion/deletion polymorphism in the gene coding for receptor associated protein (RAP) and plasma apolipoprotein AI (apoAI) and high density lipoprotein cholesterol (HDL) levels. PMID- 10845574 TI - Identification of a novel frameshift beta-thalassemia mutation in an Asian Indian. PMID- 10845575 TI - Recent developments in circadian photoreception: more than meets the eye. PMID- 10845576 TI - Human extraocular muscles: unique pattern of myosin heavy chain expression during myotube formation. AB - PURPOSE: To study the myosin heavy chain composition of the human extraocular muscles (EOMs) during development. METHODS: EOMs from human fetuses of 8 to 22 weeks of gestation were studied with immunocytochemistry and gel electrophoresis. Antibodies specific against nine isoforms of myosin heavy chain (MyHC) were used in serial frozen sections. RESULTS: The developing EOMs had a delayed time course of myotube formation and a unique composition and distribution of MyHCs compared with human limb skeletal muscle. The primary myotubes coexpressed two developmental isoforms of MyHCI from the earliest stages. The third developmental MyHCI delineated the future orbital layer at 10 to 12 weeks of gestation. MyHC slow tonic also appeared early, whereas MyHC alpha-cardiac and MyHC-extraocular, important components of adult EOM, were never detected at the gestational ages studied. CONCLUSIONS: The developmental features of the EOMs differed significantly from those reported for limb muscles of the corresponding ages. It is clear that the knowledge of limb muscle development does not fully apply to more specialized muscles, such as the eye muscles. The extreme complexity displayed by the EOMs probably reflects their distinct embryonic origin, innervation, and regulatory program of myogenesis. PMID- 10845577 TI - Immunohistochemical localization of NQO1 in epithelial dysplasia and neoplasia and in donor eyes. AB - PURPOSE: To examine the expression of NAD(P)H:quinone oxidoreductase 1 (NQO1, DT diaphorase), a potential bioactivating enzyme for mitomycin C in corneal and conjunctival epithelial dysplasia and neoplasia and in normal tissues from human donor eyes, by immunohistochemistry. METHODS: Formalin-fixed, paraffin-embedded sections of human donor eyes and tissue sections with histologic diagnoses of corneal and conjunctival epithelial dysplasia and neoplasia from the Eye Pathology Laboratory, University of Colorado Health Sciences Center were analyzed. Detection of NQO1 in tissues was performed using standard immunohistochemical techniques with monoclonal antibodies against NQO1 and immunoperoxidase staining. RESULTS: All 20 tumors stained positive for NQO1. In seven eyes from four donors, positive staining for NQO1 was detected in all epithelial and endothelial layers, in fibroblasts, in all retinal layers except the photoreceptor outer segments, and in the fascicles and arachnoid of the optic nerve. Only minimal staining was detected in the photoreceptor outer segments and the optic nerve pia and dura. Immunostaining was markedly reduced in all tissues in both eyes from donor 5. Genetic analysis confirmed that this individual was homozygous for a polymorphism in NQO1 (NQO1*2). CONCLUSIONS: NQO1 was detected by immunohistochemistry in every examined section of corneal and conjunctival epithelial dysplasia and neoplasia, suggesting that NQO1 may play a role in the bioactivation of mitomycin C in these tumors. However, the presence of NQO1 in the corneal, conjunctival, and ciliary epithelium; the retinas; and the optic nerves of donor eyes may indicate the potential for mitomycin C toxicity, particularly at higher doses. PMID- 10845578 TI - Changes in retinal and choroidal gene expression during development of refractive errors in chicks. AB - PURPOSE: During growth, the retina analyzes the projected image to achieve a close match between eye length and focal length. Because the messengers released by retina and choroid are largely unknown, genes that are differently expressed in response to changes in the retinal image were identified. In addition, because glucagon may be important in the visual control of eye growth, the transcript levels of proglucagon were studied. METHODS: Reverse transcription-polymerase chain reaction differential display was used to identify genes that were differentially expressed in chick eyes that were deprived of sharp vision or treated with positive or negative lenses. Differences were analyzed through sequencing and database searches and confirmed by Northern blot analyses. RESULTS: Combining 40 and 33 arbitrary primers with 3 oligo-dT-primers, approximately 48% and 40% of the retinal and choroidal mRNAs were screened, respectively. Twelve differences were detected in retinal tissue and five in choroidal tissue after 6 to 24 hours of exposure to defocus. Only one of 10 sequenced products could be identified as cytochrome-c oxidase, subunit I. Northern blot analysis confirmed its twofold upregulation after positive lens wear and also changes in four other unknown genes. Finally, it was shown that retinal glucagon mRNA content increased after treatment with positive lenses. CONCLUSIONS: Visual conditions that induce refractive errors produce changes in gene expression in retina and choroid within 1 day. In line with previous immunohistochemical data, it was found that the amount of glucagon mRNA was upregulated during wearing of positive lenses. PMID- 10845579 TI - Distribution of glucocorticoid and mineralocorticoid receptors and 11beta hydroxysteroid dehydrogenases in human and rat ocular tissues. AB - PURPOSE: The administration of glucocorticoids as topical or systemic medications may lead to the development of ocular hypertension through the induction of morphologic and biochemical changes in the trabecular meshwork leading to a reduction in the facility of aqueous outflow. Glucocorticoids exert their physiological effects by binding to and activating glucocorticoid and mineralocorticoid receptors. The activity of glucocorticoids is critically regulated at a prereceptor level by the two isozymes of 11beta-hydroxysteroid dehydrogenase. The purpose of this study was to determine the distribution of glucocorticoid target receptors and the isozymes of 11beta-hydroxysteroid dehydrogenase (11 beta-HSD) that regulate the activity of glucocorticoids at a prereceptor level in human and rat ocular tissues. METHODS: Horizontal sections of normal adult human and rat eyes were cut and hybridized with 35S-labeled cRNA probes specific for the glucocorticoid receptor, mineralocorticoid receptor, and 11beta-HSD types 1 and 2 using in situ hybridization. Immunohistochemical analysis of glucocorticoid and mineralocorticoid receptors using monoclonal antibodies was carried out on rat eye tissue sections. Whole rat eyes were homogenized and the activity of 11beta-HSD types 1 and 2 in the eye assessed as the percentage conversion of tritiated corticosterone to tritiated 11 dehydrocortico-sterone when corticosterone was added to the homogenate. RESULTS: In the rat ocular tissues mRNAs encoding glucocorticoid receptor, mineralocorticoid receptor, and 11beta-HSD types 1 and 2 were detected in nonpigmented ciliary epithelium, trabecular meshwork, corneal epithelium and endothelium, and anterior lens epithelium. Immunohistochemistry confirmed the presence of glucocorticoid and mineralocorticoid receptors at these sites. Activity of both isozymes of 11beta-HSD was demonstrated in homogenized rat eyes (percentage conversion of tritiated corticosterone to 11-dehydrocorticosterone; mean +/- SD, 11beta-HSD 1 = 15% +/- 5.3%, 11beta-HSD 2 = 7.9% +/- 2.8%). In both human and rat eyes, expression of mRNAs encoding glucocorticoid receptor and 11beta-HSD type 1 was high in the trabecular meshwork and lens epithelium, whereas expression of mRNAs encoding the mineralocorticoid receptor and 11beta HSD type 2 was high in nonpigmented ciliary epithelium and corneal epithelium and endothelium. CONCLUSIONS: Glucocorticoid target receptors and the enzymes regulating glucocorticoid activity at these receptors are present in mammalian ocular tissues, which regulate aqueous humor formation and outflow. Alteration in the number or affinity of receptors or in the activity of regulatory enzymes may alter the susceptibility of certain individuals to the effects of glucocorticoids on intraocular pressure. PMID- 10845580 TI - Proteoglycan composition in the human sclera during growth and aging. AB - PURPOSE: Scleral proteoglycans were characterized from human donor eyes aged 2 months to 94 years to identify age-related changes in the synthesis and/or accumulation of these extracellular matrix components. METHODS: Newly synthesized proteoglycans (previously radiolabeled with 35SO4) and total accumulated scleral proteoglycans were extracted with 4 M guanidine hydrochloride and separated by molecular sieve chromatography on a Sepharose CL-4B column. The elution positions of newly synthesized and total accumulated proteoglycans were determined by assaying each fraction for radioactivity and glycosaminoglycans, respectively. Regression analyses were performed on the three major proteoglycan peaks to identify age-related changes in scleral proteoglycan composition. Scleral proteoglycans were further purified by anion-exchange chromatography and characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analyses. RESULTS: Human scleral proteoglycans were apparent as three major peaks after chromatography on Sepharose CL-4B. The two faster eluting peaks contained alternative forms of the cartilage proteoglycan, aggrecan, whereas the third peak contained the small proteoglycans biglycan and decorin. The relative percentage of newly synthesized and total accumulated aggrecan increased approximately two- to sixfold from infancy to 94 years. In contrast, the relative percentage of newly synthesized and total accumulated biglycan and decorin decreased by approximately 25%. Chromatography and Western blot results indicated that the absolute amounts of all three proteoglycans significantly increased in concentration within the sclera from birth to the fourth decade. Beyond the fourth decade, decorin and biglycan decreased in all scleral regions and were present in lowest concentrations by the ninth decade. In contrast, aggrecan, which was present in highest concentration in the posterior sclera, was not significantly reduced with increasing age. CONCLUSIONS: The age-related changes in scleral proteoglycan composition observed in the present study are likely to contribute to the regional alterations in biomechanical properties of the sclera associated with growth and aging. PMID- 10845581 TI - Coexpression of VEGF receptors VEGF-R2 and neuropilin-1 in proliferative diabetic retinopathy. AB - PURPOSE: To elucidate vascular endothelial growth factor (VEGF)-mediated pathogenesis of fibrovascular proliferation in diabetic retinopathy. METHODS: Fibrovascular tissues were obtained at vitrectomy from 22 cases with proliferative diabetic retinopathy. The half-divided tissues were processed for reverse transcription-polymerase chain reaction (RT-PCR) analysis to examine the expression of VEGF isoforms and their receptors. Paraffin sections of the other half were used for immunohistochemistry for CD34, glial fibrillary acidic protein and VEGF, and in situ hybridization for VEGF. RESULTS: RT-PCR analysis demonstrated the expression of VEGF receptors VEGF-R1, VEGF-R2, and neuropilin-1 in 12, 14, and 14 of 22 cases, respectively. Notably, VEGF-R2 and neuropilin-1 were simultaneously expressed in the identical 14 tissues. The isoform VEGF121 was constitutively expressed in all the tissues examined, whereas the expression of VEGF165 was confined to the 7 tissues that also expressed VEGF-R2 and neuropilin-1. The vascular density of fibrovascular tissues evaluated by immunohistochemistry for CD34 was significantly higher in the cases with the expression of VEGF-R2 and neuropilin-1 than in those without their expression (P < 0.01), whereas VEGF-R1 expression had no such relationship with the vascular density. The fibrovascular tissues that expressed VEGF165 together with VEGF-R2 and neuropilin-1 were found in significantly younger patients (P < 0.01). In situ hybridization and immunohistochemical studies demonstrated that glial cells in the fibrovascular tissues express and produce VEGF. CONCLUSIONS: Coexpression of VEGF-R2 and neuropilin-1 is suggested to facilitate fibrovascular proliferation in diabetic retinopathy. PMID- 10845582 TI - Retinal ganglion cells recognized by serum autoantibody against gamma-enolase found in glaucoma patients. AB - PURPOSE: To study pathologic roles of the presence of serum autoantibodies against retinal ganglion cells in patients with glaucoma. METHODS: Serum autoantibody reactions were detected by Western blot analysis using retinal soluble fractions in 79 patients with glaucoma (normal-tension glaucoma [NTG], 23 cases; primary open-angle glaucoma [POAG], 56 cases) and 60 age-matched healthy subjects. Clinical characteristics including visual acuity, visual field, intraocular pressure (IOP), and optic disc features were compared between the serum autoantibody-positive and -negative patients. The retinal autoantigen recognized by patients' sera was identified by a combination of in-gel digestion and Edman sequencing. RESULTS: Western blot analysis revealed that serum autoantibody against retinal 50-kDa antigen was recognized in 20 out of 79 glaucoma patients (25.3%; 14 POAG and 6 NTG patients) and 60 age-matched control subjects (11.7%), respectively. Immunocytochemistry revealed that labeling of the ganglion cell layer (GCL) by IgG from glaucoma patients (POAG: 13/56, 23.2%; NTG: 6/23, 26%) existed at a significantly higher rate than that by IgG from control subjects (2/60, 3.3%; P < 0.05). In POAG, maximum IOP in the serum antibody positive-patients was significantly lower than that in the antibody-negative patients (P < 0.05). However, no statistical differences were observed in visual field loss, disc cupping, and other clinical factors between the antibody positive and -negative groups in POAG and NTG. In-gel digestion of the 50-kDa band in two-dimensional polyacrylamide gels and Edman sequence analysis of the high-performance liquid chromatography-purified peptides identified the 50-kDa protein as gamma-enolase. Injection of the 50-kDa IgG from glaucoma patients or anti-gamma-enolase serum into the vitreous cavity of Lewis rats caused reduction of the b-wave of the electroretinogram and TdT-dUTP terminal nick-end labeling (TUNEL)-positive staining within the GCL. CONCLUSIONS: In the current study, serum autoantibody against 50-kDa protein identified as gamma-enolase in 25% of glaucoma patients. PMID- 10845583 TI - Mikulicz's disease and Sjogren's syndrome. AB - PURPOSE: To characterize lacrimal gland function and lymphocyte infiltration in patients with Mikulicz's disease (MD) and Sjogren's syndrome (SS). METHODS: Four patients with MD and 5 with SS were recruited, on whom were performed Schirmer test I (Schirmer test without anesthesia), Schirmer test with nasal stimulation, and vital staining of the ocular surface. The lacrimal gland was then biopsied and the tissues stained with CD3, CD4, CD8, B220, APO2.7, Fas, and Fas ligand (Fas-L) antibodies. RESULTS: Although regular Schirmer test results in the MD group were less than 10 mm, those with nasal stimulation, 38.1 +/- 3.4 mm, were significantly greater than the SS group. There were minimal ocular surface changes in MD. Morphologic staining with hematoxylin and eosin was identical in both groups, but the acinar cells were stained with APO2.7 only in the SS group. There was strong Fas and Fas-L staining in SS patients but not in those with MD. CONCLUSIONS: Lacrimal gland acinar cells in those with MD maintained their function and were not programmed for cell death. The sicca syndrome was not observed in MD patients. Although the pathology is similar for MD and SS, the difference in acinar cell apoptosis and function can explain clinical differences. PMID- 10845584 TI - Epithelialization of a synthetic polymer in the feline cornea: a preliminary study. AB - PURPOSE: This study examined the potential of a synthetic polymer to support stable epithelial growth when implanted in the feline cornea. METHODS: A perfluoropolyether-based polymer was cast into lenticules that were coated with collagen I and implanted in four feline corneas. Epithelial growth onto the lenticules was monitored clinically for 6 weeks, after which time the animals were killed, and three corneas were evaluated histologically. Immunohistochemistry was used to identify proteins associated with the formation of a basement membrane (laminin) and adhesion complexes (bullous pemphigoid antigen and collagen VII). Electron microscopy was used to examine the tissue polymer interface for evidence of the assembly of these adhesive structures. RESULTS: Postoperative epithelial growth began on days 2 to 3, and lenticules were fully epithelialized by days 5 to 9. Lenticules were clinically well tolerated and histology showed epithelium consisting of multiple layers adherent to the lenticule's surface. Laminin, bullous pemphigoid antigen and collagen VII were identified at the tissue-polymer interface using immunohistochemistry. Ultrastructural examination showed evidence of assembly of these proteins into a recognizable basement membrane and hemidesmosomal plaques. CONCLUSIONS: A perfluoropolyether-based polymer coated with collagen I was implanted in the feline cornea and supported epithelial growth that showed signs of persistent adhesion, both clinically and histologically. This polymer shows potential for ophthalmic applications that require sustained epithelialization. PMID- 10845585 TI - Ascorbic acid content of human corneal epithelium. AB - PURPOSE: To measure the concentration of ascorbic acid in the human corneal epithelium. METHODS: Corneal epithelium was removed from postmortem eyes 4 to 16 hours after death and ascorbate measured by high-performance liquid chromatography. RESULTS: The concentration of ascorbate was 1.33 +/- 0.48 mg/gm wet weight (mean +/- SD), estimated to be 14 times its concentration in the aqueous humor. CONCLUSIONS: Ascorbate can protect the basal layer of the epithelium by absorption of incident ultraviolet radiation. PMID- 10845586 TI - Flow cytometry measurements of DNA content in primary and recurrent pterygia. AB - PURPOSE: To evaluate DNA content and cellular proliferation rates in primary and recurrent pterygia. METHODS: Matched pterygium and superior conjunctiva tissue were obtained in 36 eyes of 36 patients undergoing pterygium excision with conjunctival autografting (24 primary pterygia, 12 recurrent pterygia). Epithelial and fibrovascular layers were separated for analysis. Matched superior conjunctiva obtained at the time of surgery were used as controls. Samples were prepared according to Thompson's method, and flow cytometry was performed with a Becton-Dickinson FACScan. Analysis of histograms and calculations of cell percentages in cell cycle phases were carried out using CellFit software (version 2.0). Mean proliferation indices (MPIs) were compared using the Wilcoxon matched pair signed-rank test. RESULTS: The MPI of pterygium fibrovascular tissue (13.4) was significantly higher than the MPI of pterygium epithelium (3.1; P = 0.0001). The MPI of pterygium fibrovascular tissue was also significantly higher than that of superior conjunctival fibrovascular tissue (6.0; P = 0.0001). There was no difference in MPI values between pterygium epithelium and superior conjunctival epithelium (3.55; P = 0.12). The MPI of fibrovascular tissue from recurrent pterygium (73.75) was significantly higher than the MPI of fibrovascular tissue from primary pterygium (7.3; P = 0.003). CONCLUSIONS: The finding of high levels of cellular proliferation in the subepithelial fibrovascular layer of pterygium confirms that pterygium is a disorder of excessive cellular proliferation and that the fibrovascular layer is the site of cellular proliferation. Markedly raised levels of cellular proliferation in recurrent pterygium tissue suggest a clinical correlation between fibrovascular tissue upregulation and pterygium recurrence after surgery. PMID- 10845587 TI - Hydrogel lens monomer constituents modulate protein sorption. AB - PURPOSE: To examine the effect of hydrogel lens monomer constituents on protein sorption. METHODS: A series of hydroxyethylmethacrylate (HEMA)-based hydrogels with various amounts of methacrylic acid (MAA) or N-vinyl pyrrolidone (NVP) were synthesized. A radiolabel tracer technique was used to measure the amount of protein adsorbed on or penetrating into the hydrogels. Penetration of fluorescence-labeled proteins in the hydrogels was studied by laser scanning confocal microscopy. Single-protein solutions of human serum albumin (HSA) and hen egg lysozyme were studied. RESULTS: Inclusion of the comonomers MAA or NVP in hydrogels resulted in an increase in water content and also had a strong impact on protein sorption. An increase in the amount of MAA in the poly(HEMA-co-MAA) hydrogels increased lysozyme adsorption and penetration but reduced HSA adsorption. However, the amount of protein adsorbed for both HSA and lysozyme increased with the amount of NVP in the poly(HEMA-co-NVP) hydrogels. In contrast to the marked effect of MAA on protein sorption, in particular, on lysozyme sorption, NVP had little influence on protein sorption. When a hydrogel contains both MAA and NVP, MAA has the dominant effect on protein sorption-in particular, on lysozyme sorption. Furthermore, a large difference was observed in the amount of lysozyme adsorbed on the hydrogels that had similar water contents but little variation in adsorption of HSA. CONCLUSIONS: Negatively charged carboxyl groups of the MAA constituent may influence lysozyme sorption in two ways: by electrostatic attraction and by increasing the possibility for the small lysozyme molecule to penetrate the hydrogels. Interactions of the surface lactam groups of NVP with proteins may be attributable to the attraction of proteins to NVP. Water content is not a primary factor in determining protein adsorption. It appears that the monomer constituents, such as MAA or NVP, control protein adsorption. PMID- 10845588 TI - Increased platelet-activating factor receptor gene expression by corneal epithelial wound healing. AB - PURPOSE: Platelet activating factor (PAF) is a potent inflammatory mediator the synthesis of which increases in the cornea after injury. The effects of PAF are mediated by receptors (PAF-R), which are present in target cells. This study was undertaken to investigate the effects of wound healing, PAF, and growth factors on modulating PAF-R mRNA levels in corneal epithelial cells. METHODS: Cultures of rabbit corneal epithelial (RCE), rabbit limbal epithelial (RLE), rabbit corneal fibroblast (RCF), and rabbit corneal endothelial (RCEn) cells, as well as rabbit corneal keratocytes (RCKs) were used. For the in vivo wound-healing experiments, a 7-mm central corneal deepithelialization was performed in anesthetized rabbits. For the in vitro experiments, wounded rabbit corneas were maintained in organ culture. Corneas were stimulated with 120 nM PAF or preincubated with PAF antagonists, cyclohexamide (CHX) or actinomycin D (AcD) before adding PAF. RCE cells were stimulated with transforming growth factor (TGF)-beta1, -beta2, and, beta3, basic fibroblast growth factor (bFGF), keratinocyte growth factor (KGF); and hepatocyte growth factor (HGF). Total RNA was isolated and PAF-R expression evaluated by reverse transcription-polymerase chain reaction (RT-PCR), Northern blot analysis, and quantitative RT-PCR. RESULTS: PAF-R mRNA was expressed in RCE, RLE, and RCEn cells and RCKs, but not in RCFs. After epithelial injury, PAF-R expression increased from 2.5 to 4 times, both in vitro and in vivo. Addition of cPAF further stimulated PAF-R gene expression in epithelium, which was abolished by PAF antagonists. Quantitative RT-PCR revealed that PAF stimulated PAF-R mRNA threefold after injury. The induction of PAF-R by its agonist required previous injury and was inhibited by AcD but not by CHX. Treatment of RCE cells with TGF beta1, -beta2, or -beta3, HGF, and KGF increased mRNA in PAF-R; however, bFGF had no effect. CONCLUSIONS: Corneal injury produces changes in PAF-R mRNA expression. Whereas stroma fibroblastic cells lost the PAF-R gene expression found in keratocytes, corneal epithelial injury upregulated PAF-R mRNA. These results suggest that activation of selective growth factors and increases in PAF synthesis after injury stimulate PAF-R gene transcription and constitute important feedback mechanisms needed to maintain the inflammatory process and regulate epithelial wound healing. PMID- 10845590 TI - Clinical significance of saccade analysis in early active Graves' ophthalmopathy. AB - PURPOSE: To assess whether saccadic eye movements show distinct changes in patients with early active Graves' ophthalmopathy (GO), which could serve as a diagnostic tool for early detection and treatment. METHODS: Each of two prospective studies included 10 patients with early acute GO and 10 age- and sex matched control subjects. In the explorative study (ES) 15 dynamic parameters of saccades were analyzed. In the comparative study (CS) only those parameters were evaluated, which in ES had shown significant differences between patients and controls. Horizontal and vertical saccades of 10 degrees, 20 degrees, and 40 degrees including a fatigue test were recorded binocularly using the induction scleral search coil. RESULTS: The differences of saccadic dynamics between patients and controls were small, whereas intra- and interindividual standard deviations were large. In ES, 7.1% of the parameters showed significant differences at a level of P < or = 0.05. In CS, 2.1% of all parameters revealed repetitive significant differences. Despite statistical significance, individual data did not allow differentiation between patients and healthy individuals due to high standard deviations. CONCLUSIONS: In early active GO no clinically relevant saccadic changes were detected. These findings may be based on adaptation of the central saccadic generator. Inclusion of patients with fibrotic muscle changes due to long-standing disease could explain the contrasting results of previous studies. Consequently, analysis of saccades does not serve as a diagnostic tool during early active GO. PMID- 10845589 TI - Regulation of MMP-9 activity in human tear fluid and corneal epithelial culture supernatant. AB - PURPOSE: To evaluate human corneal epithelial culture supernatant and tear fluid for the presence of activators and inhibitors of matrix metalloproteinase (MMP) 9, MMP-3, and tissue inhibitor of metalloproteinase (TIMP)-1, respectively, and to evaluate the effect of MMP-3 on the activation of MMP-9 in these specimens. METHODS: Unstimulated tear fluid was collected from patients with ocular rosacea and normal control subjects. Levels of MMP-9, MMP-3, and TIMP-1 were determined by enzyme-linked immunosorbent assay (ELISA) and/or immunoblot analysis. Supernatants from primary human corneal epithelial cultures and human tear fluid were incubated with MMP-3. Cultured epithelial cells and their supernatants were also treated with doxycycline before MMP-3 was added. Gelatin zymography was used to identify activated 82-kDa MMP-9. MMP-9 activity was assessed with a commercial MMP-9 activity assay system. RESULTS: MMP-9 and TIMP-1 were detected at significantly higher concentrations in rosacea-affected than in normal tear fluids. MMP-3 was detected exclusively in the tear fluid of patients with ocular rosacea who had corneal epithelial disease. Treatment of the supernatant and tear fluid with MMP-3 resulted in two bands with molecular weights of 92 kDa and 82 kDa, representing pro-MMP-9 and activated MMP-9, respectively. Doxycycline added to the conditioned media did not affect activation of MMP-9 by MMP-3. However, 24 hour treatment of corneal epithelial cultures with doxycycline resulted in a lower concentration and activity of MMP-9 in their supernatants. CONCLUSIONS: MMP 9 and TIMP-1 are produced by the human corneal epithelium and are present in tear fluid. MMP-3 alone is sufficient to activate MMP-9 on the ocular surface. Doxycycline does not directly inhibit this activation by MMP-3, but it decreases MMP-9 activity when added to corneal epithelial cultures. PMID- 10845591 TI - Co-development of VEP motion response and binocular vision in normal infants and infantile esotropes. AB - PURPOSE: To determine the maturational course of nasotemporal asymmetry in infantile esotropia and to define the relationships among the symmetry of the motion visual evoked potential (MVEP), eye alignment, fusion, and stereopsis. METHODS: Sixty healthy term infants and 34 infants with esotropia participated. Nasotemporal MVEP asymmetry was assessed by the presence of a significant F1 response component with an interocular phase difference of approximately 180 degrees and by an amplitude "asymmetry index." Fusion was evaluated using the 4 p.d. base out prism test. Random dot stereoacuity was assessed in infants with forced-choice preferential looking (FPL) using the Infant Random Dot Stereocards. Eye alignment was assessed by the alternate prism and cover or the modified Krimsky test. RESULTS: Normal infants 2 to 3 months of age exhibited marked nasotemporal MVEP asymmetry, which rapidly diminished by 6 to 8 months. Neonates did not exhibit MVEP asymmetry. There was good concordance between fusion and MVEP symmetry and between stereopsis and MVEP symmetry; the concordance between MVEP symmetry and orthoposition of the visual axes was significantly poorer. The same proportion of normal and young esotropic infants showed symmetrical MVEPs. Regardless of the age at surgery, most patients with infantile esotropia had asymmetrical MVEPs after surgery. CONCLUSIONS: These data support a strong link between fusion and MVEP symmetry during both normal maturation and in infantile esotropia. Furthermore, the finding that the youngest infants with esotropia do not differ significantly from normal suggests that the nasotemporal asymmetry found in older patients with infantile esotropia does not represent an arrest of maturation but, rather, a pathologic change of the motion pathways. PMID- 10845592 TI - mRNA in situ hybridization of TIGR/MYOC in human trabecular meshwork. AB - PURPOSE: To determine the distribution of mRNA expression of the trabecular meshwork-induced glucocorticoid response protein/myocilin (TIGR/MYOC) in human trabecular meshwork. METHODS: In situ hybridization using a 1.25-kb probe obtained from reverse transcription-polymerase chain reaction of TIGR/MYOC cDNA was performed to determine the location of cell labeling within the different regions of the meshwork. The effect of dexamethasone on the pattern of labeling was studied in organ cultured meshwork. Trabecular meshwork from three sources was studied: enucleated eyes obtained at autopsy, trabeculectomy specimens obtained during filtration surgery, and meshworks from anterior segments in perfusion organ culture. Hybridization was performed on frozen sections, paraffin sections, and sections from JB-4 plastic-embedded tissue. RESULTS: Labeling for TIGR/MYOC mRNA was present in most trabecular cells of the uveal, corneoscleral, and juxtacanalicular regions but only variably present in the endothelial cells of Schlemm's canal. A similar pattern was found in the trabeculectomy specimens from eyes with primary open-angle or pseudoexfoliative glaucoma. Dexamethasone treatment increased the labeling intensity and number of labeled cells in meshwork, and also the number of labeled endothelial cells of Schlemm's canal. Fresh tissue processed within 12 hours postmortem gave more consistent labeling than older tissue, although some label was found up to 48 hours postmortem. Labeling was found in tissue from all three sources, and with all three embedding techniques; JB-4 sections provided the best morphologic resolution. CONCLUSIONS: In situ hybridization reveals that mRNA expression for TIGR/MYOC is present in most cells in all regions of the meshwork but only variably present in the endothelial cells of Schlemm's canal. Dexamethasone treatment increased the number and intensity of labeled cells, and also increased the number of labeled cells in the endothelial lining of Schlemm's canal. PMID- 10845593 TI - Automated analysis of normal and glaucomatous optic nerve head topography images. AB - PURPOSE: To classify images of optic nerve head (ONH) topography obtained by scanning laser ophthalmoscopy as normal or glaucomatous without prior manual outlining of the optic disc. METHODS: The shape of the ONH was modeled by a smooth two-dimensional surface with a shape described by 10 free parameters. Parameters were adjusted by least-squares fitting to give the best fit of the model to the image. These parameters, plus others derived from the image using the model as a basis, were used to discriminate between normal and abnormal images. The method was tested by applying it to ONH topography images, obtained with the Heidelberg Retina Tomograph, from 100 normal volunteers and 100 patients with glaucomatous visual field damage. RESULTS: Many of the parameters derived from the fits differed significantly between normal and glaucomatous ONH images. They included the degree of surface curvature of the disc region surrounding the cup, the steepness of the cup walls, the goodness-of-fit of the model to the image in the cup region, and measures of cup width and cup depth. The statistics of the parameters were analyzed and were used to construct a classifier that gave the probability, P(G), that each image came from the glaucoma population. Images were classified as abnormal if P(G) > 0.5. The probabilities assigned to each image were in most cases close to 0 (normal) or 1 (abnormal). Eighty-seven percent of the sample was confidently classified with P(G) < 0.3 or P(G) > 0.7. Within this group, the overall classification accuracy was 92%. The overall accuracy of the method (the mean of sensitivity and specificity, which were similar) in the whole sample was 89%. CONCLUSIONS: ONH images can be classified objectively and dependably by an automated procedure that does not require prior manual outlining of disc boundaries. PMID- 10845594 TI - Direct effects of muscarinic agents on the outflow pathways in human eyes. AB - PURPOSE: Recent studies demonstrating the presence of muscarinic receptors and contractile-like cells in the trabecular meshwork tissue and/or cell cultures from human eyes suggest the possibility that there may be a direct effect of muscarinic agonists on outflow facility. The present studies were conducted to determine whether muscarinic agonists could change outflow facility in perfused human ocular anterior segments, which lack an intact ciliary muscle. METHODS: Human eyes were dissected and perfused according to previously described methods. A steady state baseline facility was established for 90 minutes, after which up to four sequential concentrations ranging from 10(-9) to 10(-3) M of pilocarpine, aceclidine, or carbachol were added to the perfusion medium. In other studies, 10(-6) M atropine was perfused alone followed by 10(-7) M carbachol with 10(-6) M atropine, whereas fellow control eyes received carbachol alone. Outflow facility was measured for 60 minutes after each drug addition. The outflow facility measurement in each eye after drug administration was compared with the baseline measurement. RESULTS: Outflow facility increased from baseline facility in eyes treated with pilocarpine, aceclidine, or carbachol at lower concentrations (10( 9) to 10(-6) M) but remained unchanged at higher concentrations (10(-4) to 10(-2) M). The effects of carbachol at 10(-7) M were completely blocked by atropine. CONCLUSIONS: Muscarinic agonists increase outflow facility in human eyes by a direct stimulation of the outflow tissues in the absence of an intact ciliary muscle. This effect is biphasic, occurring at concentrations of 10(-6) M and lower with no effect at higher concentrations. PMID- 10845595 TI - Latrunculins' effects on intraocular pressure, aqueous humor flow, and corneal endothelium. AB - PURPOSE: To determine the effects of latrunculin (LAT)-A or -B on intraocular pressure (IOP), aqueous humor flow (AHF), anterior chamber (AC) protein concentration ([protein]AC), corneal endothelial permeability and morphology, and corneal thickness in living cynomolgus monkeys. METHODS: Topical LAT-A or LAT-B was administered to one eye, and vehicle to the other. IOP was measured by Goldmann tonometry, AHF and corneal endothelium transfer coefficient (ka) by fluorophotometry, [protein]Ac by Lowry assay, corneal endothelial cell morphology by specular microphotography, and corneal thickness by ultrasound pachymetry. RESULTS: LAT-A began to lower IOP at 6 hours and maximally reduced IOP by 4.6 mm Hg at 9 hours. LAT-B lowered IOP within 1 hour and maximally reduced IOP by 3.1 mm Hg at 6 hours. LAT-A increased AHF by 87% for 3 hours and increased ka by 94% over 6 hours; LAT-B increased ka by 39% over 6 hours without affecting AHF. LAT-A increased IV fluorescein entry into the cornea approximately 10 fold, but did not affect IV fluorescein entry into the AC. LAT-A increased [protein]AC by 25% at 2 hours but not 5.5 hours. LAT-B variably and insignificantly increased [protein]AC: at 1 hour but not at 6.5 hours. LAT-A induced extensive corneal endothelial pseudoguttata within 1 hour, with normal cell counts by 7 days. LAT-B increased central corneal thickness maximally by 47 microm at 3.5 hours. CONCLUSIONS: LAT-A and -B significantly reduced IOP and were consistent in their facility-increasing effect, indicating that pharmacologic disorganization of the actin cytoskeleton in the trabecular meshwork by latrunculins may be a useful antiglaucoma strategy. However, effects on corneal endothelium or ciliary epithelium are a potential safety issue. PMID- 10845596 TI - NO/cGMP pathway activation and membrane potential depolarization in pig ciliary epithelium. AB - PURPOSE: To investigate whether in isolated porcine ciliary processes, stimulation of the nitric oxide (NO)-guanylate cyclase (GC)-3',5'-cyclic guanosine monophosphate (cGMP) pathway modulates ciliary epithelial transmembrane potential. METHODS: Changes in transmembrane potential induced by the two NO donors, sodium nitroprusside (SNP; 100 microM) and S-nitroso-N-acetyl penicillamine (SNAP; 100 microM), or by the cGMP-analogue 8-para chlorophenylthioguanosine-3', 5'-cyclic guanosine monophosphate (8-pCPT-cGMP; 100 microM) were measured with microelectrodes in the presence or in the absence of the GC-inhibitor 1-H-(1,2,4)oxadiazole(4,3-alpha)quinoxalin-1-1 (ODQ; 10 microM). The effect of 8-pCPT-cGMP was also assessed in the presence of the anion channel inhibitors niflumic acid (100 microM), diisothiocyanatostilbene-2,2' disulfonic acid (DIDS; 1 mM), anthracene-9-carboxylic acid (9-AC; 1 mM), or the K+ channel blocker tetraethylammonium chloride (TEA; 10 mM). cGMP production was measured by immunoassay. RESULTS: Significant membrane depolarizations (P < 0.05-0.001; n = 5 8) were induced by SNP (6 +/- 1 mV; mean +/- SEM), SNAP (8 +/- 1 mV), or 8-pCPT cGMP (13 +/- 1 mV). In presence of ODQ, the effect of SNP and SNAP were significantly inhibited (-2 +/- 0 mV and 0 +/- 0 mV, respectively; P < 0.05; n = 5-6), but not depolarizations elicited by 8-pCPT-cGMP. These were prevented (P < 0.05-0.01; n = 5) by niflumic acid (1 +/- 1 mV), DIDS (1 +/- 1 mV), or 9-AC (5 +/ 1 mV), but not by TEA (12 +/- 2 mV). The increase in cGMP production induced by SNP (9.5-fold) was inhibited by ODQ (P < 0.001; n = 6). CONCLUSIONS: Activation of the NO-GC-cGMP pathway modulates epithelial transmembrane potential in isolated porcine ciliary processes. PMID- 10845597 TI - Ranking of optic disc variables for detection of glaucomatous optic nerve damage. AB - PURPOSE: To describe optic disc variables assessed by evaluation of clinical optic disc photographs and to compare sensitivity and specificity of these optic disc parameters in identifying patients with ocular hypertension who have nerve fiber layer defects and normal visual fields and patients with visual field defects. METHODS: The study included 500 normal subjects, 132 patients with ocular hypertension with retinal nerve fiber layer defects and normal visual fields (preperimetric glaucoma), and 840 patients with glaucomatous visual field defects. Color stereo optic disc photographs were morphometrically evaluated. RESULTS: Highest diagnostic power for the separation between the normal group and the preperimetric glaucoma group had the vertical cup-to-disc diameter ratio corrected for its dependence on the optic disc size, total neuroretinal rim area, rim-to-disc area ratio corrected for disc size, and cup-to-disc area ratio corrected for disc size. Diagnostic power was lower for rim area in the temporal inferior and temporal superior disc sector, cup area corrected for disc size, and horizontal cup-to-disc diameter ratio corrected for disc size. Less useful for the differentiation between the normal subjects and the preperimetric glaucoma group were size of zones alpha and beta of parapapillary chorioretinal atrophy, and ratios of neuroretinal rim width and rim area comparing various optic disc sectors with each other. CONCLUSIONS: In subjects with ocular hypertension with retinal nerve fiber layer defects and normal conventional achromatic visual fields, the vertical cup-to-disc diameter ratio corrected for optic disc size, total neuroretinal rim area, rim-to-disc area ratio, and cup-to-disc area ratio corrected for disc size are the most valuable optic disc variables for early detection of glaucomatous optic nerve damage. Correction for optic disc size is necessary for optic disc variables directly or indirectly derived from the optic cup. Parapapillary atrophy is less important in the early detection of glaucoma. PMID- 10845598 TI - Scaling the hill of vision: the physiological relationship between light sensitivity and ganglion cell numbers. AB - PURPOSE: Differential light sensitivity (DLS) in white-on-white perimetry is used as a measure of ganglion cell function to estimate the amount of neuronal damage in glaucoma. The physiological relationship between DLS and ganglion cell numbers is poorly understood. Within small retinal areas, brightness information is summated, so that A * L = C, or A = C/L, where A is target area, L is threshold luminance, and C is a constant. In larger illuminated areas, as with a Goldmann size III target in perimetry, summation is incomplete, so that A(k) = C/L, where k is the coefficient of summation, and 0 < k < 1. This study tests the hypothesis that the target area (A) can be represented by the number of underlying ganglion cells (G) to give G(k) = C/L. METHODS: Normative human data for ganglion cell density within 30 degrees of retinal eccentricity were taken from the literature and corrected for lateral displacement of ganglion cells from the fovea to estimate ganglion cell receptive field density (g). The number of ganglion cell receptive fields within a Goldmann size III target (G) was calculated from target area (A) and receptive field density (g) [G = A (g)]. Normative data for DLS in the central 30 degrees (Humphrey 30-2) were taken from the literature. The coefficient summation (k) was measured empirically at each Humphrey 30-2 test point in 8 normal subjects. The relationship between DLS and G was investigated by plotting DLS as decibels (dB) against G and DLS as 1/L (1/Lamberts) against G(k). The physiological relationship was extrapolated to glaucomatous ganglion cell loss by calculating hypothetical cell losses for 3 and 6 dB sensitivity defects at each test point. RESULTS: Spatial summation increased with eccentricity. The relationship between DLS (dB) and G was curvilinear. The relationship between DLS (1/L) and G(k) was linear (r2 = 0.73). The extrapolation to glaucomatous ganglion cell loss indicated that a proportionally greater loss of ganglion cells is required in the central compared with peripheral visual field for equal losses in dB sensitivity. CONCLUSIONS: The number of underlying ganglion cells, adjusted for local spatial summation, is better reflected by the DLS scale of 1/L than by dB. If spatial summation is unchanged in glaucoma, this scale more accurately reflects the amount of neuronal damage. PMID- 10845599 TI - Visual function-specific perimetry for indirect comparison of different ganglion cell populations in glaucoma. AB - PURPOSE: To compare short-wavelength automated perimetry, frequency-doubling technology perimetry, and motion-automated perimetry, each of which assesses different aspects of visual function, in eyes with glaucomatous optic neuropathy and ocular hypertension. METHODS: One hundred thirty-six eyes from 136 subjects were evaluated with all three tests as well as with standard automated perimetry. Fields were not used in the classification of study groups to prevent bias, because the major purpose of the study was to evaluate each field type relative to the others. Seventy-one of the 136 eyes had glaucomatous optic neuropathy, 37 had ocular hypertension, and 28 served as age-matched normal control eyes. Glaucomatous optic neuropathy was defined by assessment of stereophotographs. Criteria were asymmetrical cupping, the presence of rim thinning, notching, excavation, or nerve fiber layer defect. Ocular hypertensive eyes had intraocular pressure of 23 mm Hg or more on at least two occasions and normal-appearing optic disc stereophotographs. Criteria for abnormality on each visual field test were selected to approximate a specificity of 90% in the normal eyes. Thresholds for each of the four tests were compared, to determine the percentage that were abnormal within each patient group and to assess the agreement among test results for abnormality, location, and extent of visual field deficit. RESULTS: Each test identified a subset of the eyes with glaucomatous optic neuropathy as abnormal: 46% with standard perimetry, 61% with short-wavelength automated perimetry, 70% with frequency-doubling perimetry, and 52% with motion-automated perimetry. In the ocular hypertensive eyes, standard perimetry was abnormal in 5%, short wavelength in 22%, frequency doubling in 46%, and motion in 30%. Fifty-four percent (38/71) of eyes with glaucomatous optic neuropathy were normal on standard fields. However, 90% were identified by at least one of the specific visual function tests. Combining tests improved sensitivity with slight reductions in specificity. The agreement in at least one quadrant, when a defect was present with more than one test, was very high at 92% to 97%. More extensive deficits were shown by frequency-doubling perimetry followed by short-wavelength automated perimetry, then motion-automated perimetry, and last, standard perimetry. However, there were significant individual differences in which test of any given pairing was more extensively affected. Only 30% (11/37) of the ocular hypertensive eyes showed no deficits at all compared with 71% (20/28) of the control eyes (P < 0.001). CONCLUSIONS: For detection of functional loss standard visual field testing is not optimum; a combination of two or more tests may improve detection of functional loss in these eyes; in an individual, the same retinal location is damaged, regardless of visual function under test; glaucomatous optic neuropathy identified on stereophotographs may precede currently measurable function loss in some eyes; conversely, function loss with specific tests may precede detection of abnormality by stereophotograph review; and short-wavelength automated perimetry, frequency-doubling perimetry, and motion-automated perimetry continue to show promise as early indicators of function loss in glaucoma. PMID- 10845601 TI - In vitro generation of regulatory CD8+ T cells similar to those found in mice with anterior chamber-associated immune deviation. AB - PURPOSE: When injected intravenously into naive mice, peritoneal exudate cells (PECs) incubated with ovalbumin (OVA) in the presence of transforming growth factor (TGF)-beta2 induce immune deviation similar to that evoked by injection of OVA into the anterior chamber of the eye. Intraocular antigen injection elicits two distinct populations of regulatory T cells that impair delayed hypersensitivity (DH) by two different mechanisms: a CD4+ T cell that suppresses the induction of DH (afferent) and a CD8+ T cell that inhibits DH expression. In an effort to understand the origin and mechanism of action of these regulatory cells, CD8+ T cells from OVA-specific T cell receptor (Tcr) transgenic mice (OT 1) were used. METHODS: CD8+ T cells were harvested from Tcr transgenic OT-1 mice whose Tcr recognize an OVA peptide in the context of the class I major histocompatibility complex molecule Kb. These cells were stimulated in vitro with OVA-pulsed PECs exposed (or not) to TGF-beta2, then analyzed for their capacity to proliferate, to secrete various cytokines, to lyse OVA-expressing target cells, and to regulate bystander T cells in vitro and in vivo. RESULTS: When OVA pulsed PECs were used in vitro as stimulators, responding OT-1 T cells proliferated and preferentially secreted interferon (IFN)-gamma, interleukin (IL) 2, and tumor necrosis factor (TNF)-alpha, rather than IL-4 and IL-10. When the stimulator PECs were pretreated with TGF-beta2 and then pulsed with OVA, responding OT-1 T cells proliferated even more swiftly, but they secreted significantly less IFN-gamma, IL-2, and TNF-alpha, and no IL-4 or IL-10. OT-1 T cells, which constitutively display cytotoxicity toward OVA-expressing target cells, lost this activity when stimulated with OVA-pulsed, TGF-beta2-pretreated PECs. Moreover, OT-1 T cells stimulated in this manner displayed the capacity to inhibit proliferation of OVA-primed T cells exposed to OVA in vitro and to suppress in vivo the expression of OVA-triggered DH. CONCLUSIONS: OVA-pulsed PECs, pretreated with TGF-beta2, coerce naive OVA-specific CD8+ T cells to become efferent regulators of DH similar to the regulatory T cells evoked by intraocular injection of OVA. PMID- 10845600 TI - Glaucoma in primates: cytochrome oxidase reactivity in parvo- and magnocellular pathways. AB - PURPOSE: To evaluate the differential effects of ganglion cell depletion from experimental glaucoma on the relative metabolic activities of neurons in the parvo (P)- and magno (M)-cellular visual pathways of the macaque visual system. METHODS: Monocular experimental glaucoma was induced in monkeys (Macaca mulatta and M. fascicularis) by applying a laser to the trabecular meshwork to increase intraocular pressure (IOP). After other behavioral and electrophysiological studies, the lateral geniculate nuclei (LGNs) and the primary visual cortices were analyzed for functional afference from surviving ganglion cells, indicated by cytochrome oxidase (CO) histochemistry. RESULTS: CO reactivity (COR) indicated a general reduction in neural metabolism with increasing severity of glaucoma. COR in the LGNs was reduced to the same degree in both the P- and M-cellular layers. In layer 4Cbeta of the V1 cortex, the reactivity was always reduced more than in the layer 4Calpha division. CONCLUSIONS: Experimental glaucoma in monkeys reduces visual afference to the central nervous system, thereby reducing the metabolic drive as indicated by COR. The detrimental effect of glaucoma did not appear to be any greater for the M-cell, rather than the P-cell pathway in the LGN or in the visual cortex. Both are affected by the duration and severity of the experimental glaucoma. Overall, the alterations in metabolism of neurons in the parallel visual pathways supplied by the Palpha and Pbeta ganglion cells do not suggest that tests based on the functional properties of one or the other would provide optimal assessment of glaucoma. PMID- 10845602 TI - Reduced leukocyte migration, but normal rolling and arrest, in interleukin-8 receptor homologue knockout mice. AB - PURPOSE: To determine the role of the murine interleukin-8 receptor homologue (mIL-8Rh, neutrophil chemokine CXC receptor 2) in leukocyte migration using intravital microscopy in a standardized model of eye inflammation, endotoxin induced uveitis (EIU). METHODS: Two hundred fifty nanograms of E. coli endotoxin was injected into the vitreous of knockout mIL-8Rh(-/-) (n = 7) mice or heterozygous littermate mIL-8Rh(+/-) controls (n = 7). Intravital microscopic examination of iris microvasculature was performed at baseline and 6 and 24 hours after endotoxin injection. The numbers of rolling (cells/mm2 endothelial surface/min), sticking (cells/mm2 endothelial surface), and infiltrating cells (cells/mm2 iris tissue) were evaluated by digital off-line quantification. RESULTS: The number of infiltrating cells was significantly reduced in mIL-8Rh(-/ ) mice: 406 +/- 77 cells/mm2 at 6 hours and 242 +/- 50 cells/mm2 at 24 hours in mIL-8Rh(+/-) mice versus 14 +/- 4 cells/mm2 at 6 hours and 38 +/- 11 cells/mm2 at 24 hours in mIL-8Rh(-/-) mice (P < 0.001). In contrast, the absence of the IL-8 receptor homologue did not reduce rolling or sticking. CONCLUSIONS: Iris rhodamine angiography allows precise quantification of leukocyte-endothelial dynamics in the absence of surgical trauma. IL-8 and its homologues are known to be potent signals for leukocyte migration. Although IL-8 has previously been implicated in cell adhesion, video imaging in vivo demonstrated that deletion of the IL-8 receptor homologue had minimal effect on rolling or arrest in this model of inflammation. PMID- 10845603 TI - Multiplex polymerase chain reaction for diagnosis of viral and chlamydial keratoconjunctivitis. AB - PURPOSE: To develop a multiplex polymerase chain reaction (PCR) for the detection of adenovirus, herpes simplex virus, and Chlamydia trachomatis in conjunctival swabs. METHODS: Oligonucleotide primers for detection of the 3 agents were combined in one reaction and evaluated for optimal performance using control DNAs of adenovirus type 2, herpes simplex virus, and C. trachomatis plasmid. The multiplex PCR was evaluated prospectively against its corresponding uniplex PCRs, virus isolation, Chlamydia Amplicor PCR, and an immunoassay technique (immune dot blot test) in a total of 805 conjunctival swabs from patients with suspected viral and chlamydial keratoconjunctivitis. RESULTS: The multiplex PCR was as sensitive as uniplex PCRs for the detection of the agents in clinical specimens. In the prospective study, 48 of 49 (98%) clinical specimens were positive for adenovirus by the multiplex PCR compared with 26 of 49 (53%) by adenovirus isolation. For herpes simplex virus detection, the multiplex PCR had a sensitivity of 92% (34/37) compared with 94.5% (35/37) by cell culture. The multiplex PCR produced identical results to the Amplicor PCR (21/21; 100%) compared with 71% (15/21) by the immune dot blot test. CONCLUSIONS: With clinical specimens the multiplex PCR was as sensitive as its respective uniplex PCRs but more sensitive than adenovirus isolation and as sensitive as herpes simplex virus isolation or C. trachomatis Amplicor PCR. It has the potential to replace several diagnostic tests with consequent savings in cost. The test also reduces the risk of misdiagnosis by the clinicians. PMID- 10845604 TI - Recurrent intraocular inflammation in endotoxin-induced uveitis. AB - PURPOSE: Endotoxin-induced uveitis (EIU) in rats and mice peaks 24 hours after endotoxin injection and is commonly assumed to be a monophasic disease. This study examined intraocular inflammation at later time points to determine whether endotoxin injection can induce recurrent intraocular inflammation in strains of mice with high or moderate levels of susceptibility to EIU. METHODS: EIU was elicited in two mouse strains with high (C3H/HeN) and moderate (FVB/N) susceptibility, by means of intraperitoneal injections of Salmonella typhimurium endotoxin. Inflammatory cells in the anterior and posterior segments of the eye were counted by a masked observer on histologic sections of eyes from 1 to 17 days after endotoxin injection. RESULTS: A bimodal distribution of inflammatory cell infiltration was noted in eyes from C3H/HeN mice. As previously reported, inflammation peaked at 24 hours after endotoxin injection. However, a second, more pronounced peak of intraocular inflammation occurred approximately 5 days after endotoxin injection. FVB/N mice had a single peak of intraocular inflammation 4 days after injection. CONCLUSIONS: Endotoxin injection in C3H/HeN elicits recurrent intraocular inflammation. The previously unrecognized second peak of inflammation is more severe than the initial inflammatory disease. Studies on this second inflammatory peak may be useful in determining the pathogenesis of recurrent uveitis in humans. PMID- 10845605 TI - Xenoreactive CD4+ T cells and acute rejection of orthotopic guinea pig corneas in mice. AB - PURPOSE: To explore immunologic issues involved in orthotopic corneal xenotransplantation in a discordant combination using guinea pigs as donors and mice as recipients. METHODS: Two-millimeter-diameter guinea pig corneal buttons were transplanted into 1.5-mm-diameter graft beds on mouse corneas using 12 interrupted sutures. Eyelids were maintained occluded with tarsorrhaphy except at the times of clinical inspection. Grafts were considered to be rejected when the pupil margin was not visible clearly through the graft by slit-lamp microscopy. RESULTS: Guinea pig corneas protected from desiccation by persistent tarsorrhaphy survived indefinitely in the eyes of C.B-17SCID mice but were rejected acutely (but not hyperacutely) in eyes of normal BALB/c and C57BL/6 mice (median survival times, MST, 16 and 10 days, respectively). Graft survival was not extended in mice deficient in micro heavy chain or beta-2 microglobulin genes, slightly extended in mice deficient in the C3 gene (MST of 21 versus 17 days) and greatly extended in mice deficient in the CD4 gene (MST of 26 versus 9 days). Reconstitution of CD4 knock-out (KO) mice with CD4+ T cells promoted acute rejection of corneal xenografts. CONCLUSIONS: Hyperacute rejection does not occur in guinea pig corneal xenografts in mouse eyes, indicating that corneal xenografts are less vulnerable to this type of rejection than other solid tissue xenografts. CD4+ T cells are the primary mediators of acute graft rejection, although complement may contribute in a minor way. Neither antibodies nor CD8+ T cells participate in acute graft rejection. Because guinea pig cornea grafts in eyes of CD4KO mice are rejected in a delayed fashion, other innate and/or adaptive immune effectors must also be able to cause rejection of orthotopic corneal xenografts. PMID- 10845606 TI - Inhibition of FGF-induced alphaA-crystallin promoter activity in lens epithelial explants by TGFbeta. AB - PURPOSE: Fibroblast growth factor (FGF) plays a key role in normal lens biology, and recent studies suggest that transforming growth factor (TGF)-beta is involved in the origin of certain forms of cataract. In the current study, the effects of FGF and TGFbeta on alphaA-crystallin promoter activity were investigated. METHODS: Rat lens epithelial explants were cultured with or without growth factors after transfecting with the firefly luciferase reporter gene driven by either the mouse alphaA-crystallin promoter region or a control simian virus (SV)40 promoter. RESULTS: FGF-2, at a concentration that induced lens fiber differentiation, strongly stimulated alphaA-crystallin promoter activity in explants at 3 to 4 days of culture, whereas SV40 promoter control specimens showed no comparable increase. At lower concentrations of FGF, sufficient to induce cell proliferation but not differentiation, there was only a slight increase in alphaA-crystallin promoter activity. Stimulation of alphaA-crystallin promoter activity induced by the fiber-differentiating concentration of FGF was virtually abolished by as little as 25 pg/ml TGFbeta2, but the onset of fiber specific beta-crystallin accumulation was not prevented at this concentration. Phase-contrast microscopy revealed overt cataractous changes only at concentrations of TGFbeta more than 25 pg/ml. CONCLUSIONS: The stimulation of alphaA-crystallin promoter activity by FGF is consistent with its role in inducing accumulation of crystallins in explants. The blocking effect of TGFbeta on this process, even at a concentration too low to induce obvious pathologic changes, indicates the potential for TGFbeta to disturb alphaA-crystallin gene expression during early fiber differentiation. PMID- 10845607 TI - Overexpression of the transforming growth factor-beta-inducible gene betaig-h3 in anterior polar cataracts. AB - PURPOSE: In anterior polar cataracts and the fibrosis that can occur after cataract surgery, lens epithelial cells synthesize abundant extracellular matrix molecules and transdifferentiate into myofibroblast-like cells. Transforming growth factor (TGF)-beta has been implicated as a key player in these cataractous changes. The purpose of this study was to determine whether the TGF-beta inducible gene h3 (betaig-h3) is expressed in lens epithelial cells from patients with anterior polar cataracts and to test whether betaig-h3 is induced by TGF beta in cultured lens epithelial cells. METHODS: Lens epithelial cells attached to the anterior capsules of human cataractous lenses and noncataractous lenses were examined for the expression of betaig-h3 mRNA and protein using reverse transcription-polymerase chain reaction and immunohistochemical analyses. The effect of TGF-beta on betaig-h3 gene expression was also tested in human lens epithelial B-3 cells using Northern and Western blot analyses. RESULTS: betaig-h3 mRNA was not detected in lens epithelial cells from patients with clear lenses or patients with nuclear cataracts. Significant expression of mRNA for betaig-h3 was observed in lens epithelial cells from patients with anterior polar cataracts. Immunohistochemical analysis using anti-betaig-h3 antiserum indicated that betaig h3 protein was present within the subcapsular plaques of anterior polar cataracts. Treatment of human lens epithelial B-3 cells with TGF-beta1 led to an increase in betaig-h3 mRNA and the secretion of betaig-h3 protein into the culture medium. CONCLUSIONS: betaig-h3 may serve as a marker for anterior polar cataracts in addition to previously known proteins, fibronectin, type I collagen, and alpha-smooth muscle actin. The functions of this protein in lens pathology need to be further investigated. PMID- 10845608 TI - Steroid-induced short term diabetes in chick embryo: reversible effects of insulin on metabolic changes and cataract formation. AB - PURPOSE: To determine the reversible effect of insulin on glucocorticoid (GC) induced cataract formation in relation to systemic metabolic changes in the developing chick embryo. METHODS: Hydrocortisone sodium succinate (HC; 0.25 micromoles) was administered to 15-day-old embryos followed by administration of long-acting recombinant human insulin, 4 and 28 hours later. At the indicated time after HC administration, the incidence of cataractous lenses and any changes in the components of the lenses, liver, and blood were determined. RESULTS: At 48 hours after HC administration, the following observations were made: opacification of lenses; an elevation of glucose and lipids in the blood and lenses; an increase in lipid peroxide (LPO) in the blood, liver, and lenses; a decrease in glutathione (GSH) in the lens and liver (at 24 hours after HC administration); and a depletion of adenosine triphosphate (ATP) in the liver. These changes in response to HC administration were reversed by a double application of insulin. CONCLUSIONS: Insulin antagonizes GC-induced gluconeogenesis, stimulates glycolysis, and ultimately leads to recovery of decreased activity in the citric acid cycle. The restoration of ATP by the recovered citric acid cycle may facilitate de novo synthesis of GSH, which in turn may diminish GC-induced elevation of LPO in the liver. Thus, the metabolic changes in response to HC-accelerated gluconeogenesis in the liver, which can be reversed by insulin, are likely to produce oxidative stress that leads to cataract formation. GC-induced metabolic changes in the liver, which are antagonized by insulin, may relate to production of one of the risk factors for cataract formation. PMID- 10845609 TI - Chloride secretion by bovine ciliary epithelium: a model of aqueous humor formation. AB - PURPOSE: To study the physiological mechanisms of the Cl transport across the bovine ciliary body-epithelium (CBE). METHODS: Fresh isolated bovine CBE was mounted in an Ussing-type chamber. The effects of ion substitution and transport inhibitors on electrical measurements and Cl transport were investigated. RESULTS: The potential difference (PD) across the preparation was 0.55 +/- 0.04 mV and was consistently negative at the aqueous side. The short-circuit current (SCC) and tissue resistance (Rt) were found to be 8.0 +/- 0.7 microA/cm2 and 72 +/- 3 omega/cm2, respectively. Both the PD and the SCC of the bovine CBE were found to depend primarily on the concentration of the Cl bath and to a lesser extent on the Na or HCO3 concentration. At 30 mM Cl, the polarity of the PD and the direction of the SCC were reversed. Reducing the extracellular Na or Cl concentration abolished the net Cl transport into the eye under the short circuited condition. Bilateral bumetanide (0.1 mM), but not 4,4' diisothiocyanatostilbene-2-2'-disulfonic acid (DIDS; 0.1 mM), greatly inhibited the Cl transport. Bumetanide, when applied to either side, inhibited the Cl transport. The effect, however, was three times greater on the stromal side than on the aqueous surface. Bilateral heptanol (3.5 mM) and 5-nitro-2-(3 phenylpropylamino)-benzoic acid (NPPB; 0.1 mM) on the aqueous side also inhibited the Cl transport by 80% and 92%, respectively. CONCLUSIONS: The results suggest that the major Cl influx pathway at the basolateral border in pigmented epithelial cells is through the Na-K-2Cl cotransporter, but not the Cl/HCO3 and Na/H double exchangers. Intercellular gap junctions between the two cell layers and the NPPB-sensitive Cl channels at the basolateral surface in nonpigmented epithelial cells also play a crucial role in regulating the Cl movement across the functional syncytium. PMID- 10845610 TI - Mechanisms mediating substance P-induced contraction in the rat iris in vitro. AB - PURPOSE: To determine some of the mechanisms by which substance P (SP) induces contraction in the isolated rat iris. METHODS: Rings of rat iris were mounted in a 5-ml organ chamber containing Krebs solution at 37 degrees C under basal tension of 75 mg, and isometric tension was recorded. RESULTS: Substance P produced graded contraction in the rat iris, being approximately 40-fold more potent than carbachol. Peptidase inhibitors (captopril, phosphoramidon, thiorphan) did not affect the SP response. The SP contraction was dependent on external Ca2+ by a mechanism resistant to both nifedipine and omega-conotoxin GVIA. Atropine and tetrodotoxin significantly shifted the SP response to the right (three- and fivefold, respectively). Neither phorbol nor genistein altered the SP-induced contraction, whereas staurosporine caused a weak inhibition. Indomethacin, pyrilamine, guanethidine, 8-37 calcitonin gene-related peptide (CGRP) fragment, and NG-nitro-L-arginine methyl ester had no effect on SP response. All the natural tachykinin agonists caused concentration-dependent contraction in rat iris with similar maximal responses. The NK3 selective agonist senktide caused graded contraction, being approximately 150-fold more active than the NK2 selective agonist [beta-ala] NKA. The NK1 selective agonist SP methyl ester induced a small contraction. The NK3 and NK2 antagonists SR 142801 and SR 48968 shifted the SP response to the right. Schilds plots gave pA2 (negative logarithm of the molar concentration of antagonist causing a twofold rightward displacement of the concentration response curves) values of 9.37 and 7.97 and slopes of 0.70 and 1.02, respectively. CONCLUSIONS: Substance P produces a potent contraction in the isolated rat iris that seems to depend on the neural release of acetylcholine by tetrodotoxin-sensitive mechanisms. Its response relies largely on external Ca2+, through mechanisms independent of activation of L- or N type Ca2+ channels, and is probably mediated via activation of NK3 and NK2 receptors. PMID- 10845611 TI - Pressure-induced myogenic responses in isolated bovine retinal arteries. AB - PURPOSE: To investigate whether a pressure-induced myogenic vasoconstriction can be demonstrated in isolated bovine retinal arteries and to determine the cellular mechanisms involved. METHODS: Isolated bovine retinal arteries were mounted on a pressure myograph without flow and exposed to stepwise increases in intraluminal pressure. Changes in internal diameter were monitored continuously using an inverted microscope video system. RESULTS: Bovine retinal arteries showed myogenic tone at pressures higher than 10 mm Hg. This pressure-induced contraction was absent in calcium-free Krebs-Ringer bicarbonate solution. Inhibition of L-type voltage-operated calcium channels with nifedipine (1 microM) suppressed the myogenic contraction. After depolarization of the vascular smooth muscle cells with a K+ 120 mM solution, a pressure-induced contraction was still observed, indicating that besides stimulation of voltage-operated calcium channels, depolarization-independent mechanisms contribute to the pressure induced myogenic vasoconstriction. CONCLUSIONS: Isolated bovine retinal arteries spontaneously contract when exposed to raised intraluminal pressure. This response depends on extracellular calcium and is blocked by nifedipine. In addition, depolarization-independent mechanisms seem to be involved. PMID- 10845612 TI - Enhancement of glucose transport by vascular endothelial growth factor in retinal endothelial cells. AB - PURPOSE: To investigate effects of vascular endothelial growth factor (VEGF) on glucose transport and GLUT1 glucose transporter expression in primary bovine retinal endothelial cell (BREC) cultures. METHODS: Glucose transport in control and VEGF-treated BREC cultures was determined by measurement of [14C]-3-O methylglucose (3MG) uptake. GLUT1 protein and mRNA was determined by Western and Northern blot analyses, respectively. Protein kinase C (PKC) activity was measured in control and VEGF-treated cultures, and glucose transport was determined with and without prior PKC depletion and PKC inhibition. RESULTS: Dose dependent increases in 3MG uptake were seen in the VEGF-treated cultures, with an increase of 69% after a 24-hour exposure to 50 ng/ml VEGF (P < 0.001). Total cellular GLUT1 mRNA or protein, however, was unchanged. Western blot analysis of plasma membrane fractions revealed a 75% increase in plasma membrane GLUT1 in VEGF-treated cultures (P = 0.02), suggesting that the VEGF-stimulated increase in glucose transport was due to a translocation of GLUT1 to the cell membrane. VEGF stimulated a 90% increase in PKC activity in membrane fractions from cultures treated with VEGF, and VEGF-stimulated enhancement of glucose transport was abolished by cellular PKC depletion and by general and PKC beta inhibition. CONCLUSIONS: The present study demonstrates VEGF-mediated enhancement of retinal endothelial cell glucose transport and suggests that this increase is due to PKC beta-mediated translocation of cytosolic GLUT1 to the plasma membrane surface. Upregulation of retinal endothelial cell glucose transport by various factors associated with the development of retinopathy may be responsible for the metabolic derangements observed in the diabetic inner blood-retinal barrier in vivo. PMID- 10845613 TI - Mechanisms of hepatocyte growth factor-induced retinal endothelial cell migration and growth. AB - PURPOSE: Hepatocyte growth factor (HGF), also called scatter factor, stimulates growth and motility in nonocular endothelial cells and smooth muscle cells through its receptor c-Met. Recent reports suggest that HGF is increased in the serum and vitreous of patients with proliferative diabetic retinopathy and that smooth muscle cells and retinal pigment epithelial cells secrete HGF in the eye. However, little is known about HGF's action in the retina. In this study, the activity, expression, and signaling pathways of HGF were investigated in bovine retinal microvascular endothelial cells (BRECs). METHODS: Mitogenic and motogeneic effects of HGF on BRECs were examined using cell counts, thymidine uptake, and migration assays. MAP kinase (MAPK) phosphorylation was examined by Western blot analysis. Protein kinase C (PKC), MAPK, and PI3 kinase involvement were evaluated using selective inhibitors and activity assays. Expression of HGF and c-Met was evaluated by reverse transcription-polymerase chain reaction. RESULTS: HGF and c-Met were both expressed in BRECs. HGF stimulated BREC growth in a time- and dose-dependent manner, observed at HGF concentrations of 5 ng/ml or more and maximal (410%) at 100 ng/ml (P < 0.001). HGF increased BREC migration in a dose-dependent manner with a maximal 3.4-fold increase at 50 ng/ml after 5 hours. HGF induced time- and dose-dependent MAPK phosphorylation, initially evident at 5 minutes (P < 0.001) or 5 ng/ml (P < 0.050) and maximal after 15 minutes (>80-fold, P < 0.001) or 50 ng/ml (>20-fold, P < 0.001), respectively. MAPK phosphorylation was maintained for more than 2 hours. This response was inhibited 31% by 0.1 microm wortmannin and 76% by 30 microm LY294002, another PI3 kinase inhibitor. The non-isoform-selective PKC inhibitor GFX inhibited HGF induced MAPK phosphorylation by only 15% at 5 microm. Combined PKC and PI3 kinase inhibition was additive (P < 0.05). Cell migration was inhibited 30% by wortmannin (P < 0.01) and 32% by GFX (P < 0.05), and again the effect was additive (P < 0.001). HGF-induced BREC growth was suppressed by PI3 kinase, PKC, or MAPK inhibition (all P < 0.01). HGF (50 ng/ml) stimulated PI3 kinase activity 347% (P < 0.001) and PKC activity 37% (P < 0.05). HGF-induced MAPK phosphorylation and mitogenesis were not inhibited by vascular endothelial growth factor (VEGF)-neutralizing antibody. CONCLUSIONS: HGF and its receptor are expressed in BREC, and HGF stimulates both BREC growth and migration at concentrations observed in the human eye with diabetic retinopathy. HGF signaling appears to involve activation of both PKC and PI3 kinase, inducing MAPK phosphorylation that is critical for migration and growth. However, VEGF does not appear to mediate these initial HGF effects. These results indicate that HGF could have a significant role in mediating retinal endothelial cell proliferation and migration in diabetic retinopathy, and they begin to elucidate the signal transduction pathway by which this action may occur. PMID- 10845614 TI - A frequent 1085delC/insGAAG mutation in the RDH5 gene in Japanese patients with fundus albipunctatus. AB - PURPOSE: To identify the frequency of a mutation of the RDH5 gene in Japanese patients with hereditary retinal degeneration and to characterize clinical findings for the patients associated with a 1085delC/insGAAG mutation in the RDH5 gene. METHODS: Mutation screening by single-strand conformation polymorphism was performed on 6 patients with fundus albipunctatus and 150 patients with autosomal recessive retinitis pigmentosa. The DNA fragment that showed abnormal mobility on SSCP was then sequenced. Clinical features were characterized by visual acuity, slit-lamp biomicroscopy, electroretinography, fluorescein angiography, kinetic visual field testing, and dark adaptometry. RESULTS: A novel 1085delC/insGAAG mutation in the RDH5 gene was identified in all 6 patients, from 4 unrelated families with fundus albipunctatus. The ophthalmic findings of each affected member were very similar, which may provide the natural course of the phenotype produced by the 1085delC/insGAAG mutation. CONCLUSIONS: A homozygous1085delC/insGAAG mutation in the RDH5 gene produces fundus albipunctatus in Japanese patients. These findings suggest that this mutation was a founder effect in Japanese patients with fundus albipunctatus. PMID- 10845615 TI - Disease expression of RP1 mutations causing autosomal dominant retinitis pigmentosa. AB - PURPOSE: To determine the disease expression in heterozygotes for mutations in the RP1 gene, a newly identified cause of autosomal dominant retinitis pigmentosa (adRP). METHODS: Screening strategies were used to detect disease-causing mutations in the RP1 gene, and detailed studies of phenotype were performed in a subset of the detected RP1 heterozygotes using electroretinography (ERG), psychophysics, and optical coherence tomography (OCT). RESULTS: Seventeen adRP families had heterozygous RP1 changes. Thirteen families had the Arg677ter mutation, whereas four others had one of the following: Pro658 (1-bp del), Ser747 (1-bp del), Leu762-763 (5-bp del), and Tyr1053 (1-bp del). In Arg677ter RP1 heterozygotes, there was regional retinal variation in disease, with the far peripheral inferonasal retina being most vulnerable; central and superior temporal retinal regions were better preserved. The earliest manifestation of disease was rod dysfunction, detectable as reduced rod ERG photoresponse maximum amplitude, even in heterozygotes with otherwise normal clinical, functional, and OCT cross-sectional retinal imaging results. At disease stages when cone abnormalities were present, there was greater rod than cone dysfunction. Patients with the RP1 frameshift mutations showed similarities in phenotype to those with the Arg677ter mutation. CONCLUSIONS: Earliest disease expression of RP1 gene mutations causing adRP involves primarily rod photoreceptors, and there is a gradient of vulnerability of retinopathy with more pronounced effects in the inferonasal peripheral retina. At other disease stages, cone function is also affected, and severe retina-wide degeneration can occur. The nonpenetrance or minimal disease expression in some Arg677ter mutation-positive heterozygotes suggests important roles for modifier genes or environmental factors in RP1 related disease. PMID- 10845616 TI - Hemangioblastomas of the retina: impact of von Hippel-Lindau disease. AB - PURPOSE: To assess the prevalence of von Hippel-Lindau (VHL) disease and prognosis of vision in patients with retinal hemangioblastomas (HBs). METHODS: Thirty-six consecutive patients with retinal HBs were treated at Helsinki University Hospital between 1974 and 1998. Detailed neurologic, ophthalmologic, and radiologic examinations; pedigree; mutation analyses; and collection of all relevant clinical, imaging, operative, and autopsy data were performed to identify VHL. RESULTS: The median follow-up time was 10 years. No patient was lost to follow-up. There were three patient groups: 1) 11 patients with clinically definite VHL; 2) 10 patients with clinically suspected VHL with more than one retinal HB (5/10) or visceral cysts (5/10), but with no family history, no detected germ-line mutations, and no VHL-related neoplasms; and 3) 15 patients without VHL with a single retinal HB but no other data suggestive of VHL. In the 11 patients with definite VHL, retinal HBs were detected at a median age of 27 years versus 40 years in the 15 non-VHL patients, and 21 of the 22 eyes were affected. Two VHL patients were totally blind at the end of follow-up compared with one legally blind patient with suspected VHL, but none of the non-VHL patients was blind. The clinical appearance of HBs did not differ among the patient groups. CONCLUSIONS: The prevalence of VHL among patients with retinal HBs was 30% to 58% (11-21 of 36). Visual prognosis was more favorable in non-VHL than VHL patients. All patients with retinal HB should undergo thorough VHL exclusion. PMID- 10845617 TI - Methods and limits of digital image compression of retinal images for telemedicine. AB - PURPOSE: To investigate image compression of digital retinal images and the effect of various levels of compression on the quality of the images. METHODS: JPEG (Joint Photographic Experts Group) and Wavelet image compression techniques were applied in five different levels to 11 eyes with subtle retinal abnormalities and to 4 normal eyes. Image quality was assessed by four different methods: calculation of the root mean square (RMS) error between the original and compressed image, determining the level of arteriole branching, identification of retinal abnormalities by experienced observers, and a subjective assessment of overall image quality. To verify the techniques used and findings, a second set of retinal images was assessed by calculation of RMS error and overall image quality. RESULTS: Plots and tabulations of the data as a function of the final image size showed that when the original image size of 1.5 MB was reduced to 29 KB using JPEG compression, there was no serious degradation in quality. The smallest Wavelet compressed images in this study (15 KB) were generally still of acceptable quality. CONCLUSIONS: For situations where digital image transmission time and costs should be minimized, Wavelet image compression to 15 KB is recommended, although there is a slight cost of computational time. Where computational time should be minimized, and to remain compatible with other imaging systems, the use of JPEG compression to 29 KB is an excellent alternative. PMID- 10845618 TI - Significant reduction of the panretinal oxygenation response after 28% supplemental oxygen recovery in experimental ROP. AB - PURPOSE: To test the hypothesis that after supplemental oxygen recovery (SOR) in the newborn rat model of retinopathy of prematurity (ROP) the preretinal neovascular (NV) incidence and severity are decreased and the panretinal oxygenation ability is improved. METHODS: Newborn rats were first raised in either room air (controls) or variable oxygen (50%/10%) for 14 days. The experimental rats were recovered during the next 6 days (until day 20) in either room air (21% O2) or supplemental oxygen (28%). All groups were then exposed to room air for an additional 6 days (until day 26). On day 20, magnetic resonance imaging (MRI) was used to determine the panretinal oxygenation response (deltaPO2, mm Hg) to a carbogen (95% O2/5% CO2) inhalation challenge. On days 20 and 26, the retinas from a different subset of control, room air-recovered, or SOR-recovered animals were analyzed using ADPase stained or fluorescein-labeled dextran infused retinal flatmounts. RESULTS: On day 20, the panretinal deltaPO2 of the room air-recovered group (125 +/- 5 mm Hg, mean +/- SEM, n = 12) was significantly (P < 0.05) lower than that of the control group (179 +/- 6 mm Hg, n = 11). The panretinal deltaPO2 value for the SOR group (87 +/- 5 mm Hg, n = 7) was significantly (P < 0.05) lower than both the room air-recovered group and the control group. The NV incidence and severity were significantly reduced (P < 0.05) in the SOR animals compared with the room air-recovered animals. In contrast, on day 26 (after 6 days in room air), the NV incidence was statistically (P < 0.05) greater in the animals that had been exposed to SOR compared with room air-recovered animals. CONCLUSIONS: After 28% SOR, the expected decrease in NV incidence and severity occurred but with an unexpected decrease in panretinal oxygenation ability. The present data strongly support an association between subnormal panretinal oxygenation ability and increased NV risk in the newborn rat ROP model. MRI appears to be a powerful new approach for quantitatively and noninvasively measuring retinal oxygenation and may be applicable to study other ischemic or ischemia-related retinopathies in addition to ROP, such as diabetic retinopathy, sickle cell retinopathy, macular degeneration, and glaucoma. PMID- 10845619 TI - An abnormal response of retinoblastoma cells (Y-79) to neurotrophins. AB - PURPOSE: To clarify the expression of neurotrophins and their receptors in retinoblastoma (Rb) cells, to elucidate their potential role in the proliferation of neuroectodermal tumor cells, and to establish conditions for Rb cell differentiation. METHODS: The Rb-derived cell line Y-79 was grown in serum-free suspension or monolayer culture. Proliferating and differentiated cells were isolated and submitted to semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) analysis, immunostaining, and flow cytometry. The proliferation rate of the cells was estimated by 5-bromo-2'-deoxyuridine (BrdU) incorporation, and the effects of neurotrophins and laminin on BrdU-incorporation, process outgrowth, or immunostaining were determined. RESULTS: In contrast to previously studied normal retinal precursor cells, Y-79 cells not only express nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), and p75, but also the corresponding high affinity receptors TrkA, TrkB, and TrkC. Proliferation was stimulated by exogenous and endogenous neurotrophin receptor ligands. Inhibition of protein kinase phosphorylation with K252a blocked proliferation and promoted differentiation. The effect of K252a on differentiation was enhanced by the addition of soluble laminin. After 9 days of combined treatment, the fraction of differentiated cells amounted to 30%, differentiation being characterized by improved attachment, neurite outgrowth, expression of NF-68, and a loss of glial fibrillary acidic protein (GFAP) and parvalbumin immunoreactivity. These changes were accompanied by a downregulation of TrkB and TrkC, but not TrkA or p75. Differentiated cells were isolated and further grown in the absence of K252a. However, despite the high level of TrkA expression in differentiated cells, the addition of NGF had no effect on their survival. CONCLUSIONS: A mitogenic action of neurotrophins could contribute to retinal tumor growth. PMID- 10845620 TI - Concurrent downregulation of a glutamate transporter and receptor in glaucoma. AB - PURPOSE: Elevated levels of extracellular glutamate have been implicated in the pathophysiology of neuronal loss in both central nervous system and ophthalmic disorders, including glaucoma. This increase in glutamate may result from a failure of glutamate transporters, which are molecules that ordinarily regulate extracellular glutamate. Elevated glutamate levels can also lead to a perturbation in glutamate receptors. The hypothesis for the current study was that glutamate transporters and/or receptors are altered in human glaucoma. METHODS: Immunohistochemical analyses of human eyes with glaucoma and control eyes were performed to evaluate glutamate receptors and transporters. These molecules were also assayed in rat eyes injected with glial-derived neurotrophic factor (GDNF). RESULTS: Glaucomatous eyes had decreased levels of both the glutamate transporter, excitatory amino acid transporter (EAAT)-1, and the glutamate receptor subunit N-methyl-D-aspartate (NMDA)-R1. Eyes treated with GDNF had elevated levels of both EAAT1 and NMDAR1. CONCLUSIONS: The loss of EAAT1 in glaucoma may account for the elevated level of glutamate found in glaucomatous vitreous and lead to a compensatory downregulation of NMDAR1. Inasmuch as GDNF can increase levels of both EAAT1 and NMDAR1, it may be a useful therapeutic approach to restore homeostatic levels of these in glaucoma. PMID- 10845621 TI - High levels of E-/P-cadherin: correlation with decreased apical polarity of Na/K ATPase in bovine RPE cells in situ. AB - PURPOSE: The adherens junction protein E-cadherin induces a basolateral polarity of Na/K ATPase in most epithelial cells that express it, whereas in retinal pigment epithelium (RPE) cells, Na/K ATPase is largely apical. The purpose of this study was to determine whether the distribution of Na/K ATPase differs in RPE cells in situ, that differ in levels of junctional E-cadherin. METHODS: Bovine RPE cells in situ were immunostained with an E-cadherin antibody (which has some cross-reactivity with the closely related epithelial cadherin P cadherin), and RPE cells with different levels of junctional stain were identified. RPE cells with low and high E-/P-cadherin were costained in various combinations with Na/K ATPase and interacting proteins of the membrane cytoskeleton (ankyrin, fodrin, and actin) and analyzed by confocal imaging. RESULTS: Individual RPE cells within the same monolayer differed in amount of Na/K ATPase, with a lower frequency of high expressing cells in the area centralis. High expressing Na/K ATPase cells were found among cells with both low and high E-/P-cadherin levels. In cells with low E/P-cadherin, Na/K ATPase localized to apical microvilli, whereas in high E-/P-cadherin cells, Na/K ATPase was on basolateral surfaces in addition to microvilli. Actin staining showed that microvillar domains were smaller and that lateral membrane domains were taller in high E-/P-cadherin cells. In high but not low E-/P-cadherin cells, ankyrin and fodrin levels varied among cells, with a subset of cells showing distinctly higher expression. Both ankyrin and fodrin had complex subcellular distribution patterns, although they tended to be enriched basal to rather than apical to the adherens junction. Cells with high Na/K ATPase did not necessarily have commensurately higher levels of ankyrin or fodrin. Where both Na/K ATPase and ankyrin were high, they codistributed weakly in apical microvilli but more prominently on the basal cell surface. CONCLUSIONS: Within the same RPE monolayer, the polarity of Na/K ATPase differs among cells, with a more basal polarity found in cells with high levels of junctional E-/P-cadherin. The increased basal Na/K ATPase was due to a combination of a smaller microvillar domain, a taller lateral domain, and more basolateral staining for Na/K ATPase, perhaps because of an enrichment of a basal ankyrinfodrin membrane cytoskeleton with which Na/K ATPase is known to associate. PMID- 10845622 TI - Synergistic effect of Bcl-2 and BAG-1 on the prevention of photoreceptor cell death. AB - PURPOSE: Ectopic expression of Bcl-2 in photoreceptors of mice with retinal degenerative disease slows progression of the disease. BAG-1 has previously been shown to augment the inhibitory effect of Bcl-2 on programmed cell death in cultured cell systems. This study was designed to determine whether the coexpression of BAG-1 and Bcl-2 in the photoreceptors of mice with an autosomal dominant form of retinitis pigmentosa (RP) would enhance the protective effect provided by Bcl-2 alone. METHODS: An expression vector using the 5' regulatory region of the murine opsin gene was used to target the expression of BAG-1 specifically to photoreceptor cells of mice. The BAG-1 transgenic mice were crossed to Bcl-2 transgenics to obtain animals that coexpress the two transgenes in photoreceptor cells. BAG-1/Bcl-2 animals were then crossed to an RP mouse model (a transgenic line overexpressing the S334ter rhodopsin mutant) to assess the effect of coexpression of BAG-1 and Bcl-2 on retinal degeneration. Morphologic analysis was performed on retinas isolated at various times after birth to monitor disease progression. RESULTS: High levels of BAG-1 expression resulted in retinal degeneration that was not prevented by Bcl-2 expression. However, coexpression of appropriate levels of BAG-1 and Bcl-2 was found to have a profound inhibitory effect on retinal degeneration caused by overexpression of a mutant rhodopsin transgene. Whereas expression of Bcl-2 alone was previously found to delay degeneration of the retina from 2 weeks to approximately 4 weeks of age, coexpression of BAG-1 and Bcl-2 inhibited photoreceptor cell death for as long as 7 to 9 weeks. CONCLUSIONS: The synergistic effect against photoreceptor cell death produced by the coexpression of Bcl-2 and BAG-1 indicates that these proteins can function in concert to prevent cell death. At the correct dosage, coexpression of Bcl-2 and BAG-1 may serve as a potential means to treat retinal degenerative diseases. PMID- 10845623 TI - Adrenomedullin in cultured human retinal pigment epithelial cells. AB - PURPOSE: To determine whether adrenomedullin (ADM), a vasorelaxant peptide is produced and secreted by human retinal pigment epithelial (RPE) cells, whether ADM expression is regulated by inflammatory cytokines and a growth factor, and whether ADM has proliferative effects on these cells. METHODS: Production and secretion of ADM by cultured human RPE cells were examined by Northern blot analysis and radioimmunoassay. Regulation of the ADM expression by basic fibroblast growth factor, interferon (IFN)-gamma, tumor necrosis factor-alpha, interleukin (IL)1beta, or all-trans-retinoic acid was studied. In addition, proliferative effects of ADM on human RPE cells were examined by modified 3-(4,5 dimetylthiazol-2-yl)2,5-diphenyltetrazolium bromide (MTT) assay. RESULTS: ADM mRNA was expressed constitutively in all three human RPE cell lines (F-0202, D407, and ARPE-19) examined. Immunoreactive ADM was detected in the cultured media by radioimmunoassay. Sephadex G-50 column chromatography of the cultured medium showed a single peak eluting in the position of ADM-(1-52). Treatment with IFN-gamma or IL-beta increased ADM mRNA levels and immunoreactive-ADM levels in the medium in dose- and time-dependent manners in ARPE-19 cells. Exogenously added ADM increased the number of F-0202 cells and ARPE-19 cells, and the treatment with ADM antibody or ADM-(22-52) (an ADM antagonist) decreased it. CONCLUSIONS: Human RPE cells produced and secreted ADM. IFN-gamma and IL-1beta induced ADM expression in ARPE-19 cells. Furthermore, ADM stimulated proliferation of RPE cells. These results raise the possibility that ADM is related to the pathophysiology of some inflammatory and proliferative ocular diseases. PMID- 10845624 TI - Glial reactivity, an early feature of diabetic retinopathy. AB - PURPOSE: To characterize early structural gliotic reactions in retinal Muller cells, astrocytes, and microglia in experimentally induced diabetes. METHODS: Rats were rendered diabetic by streptozotocin injection and killed after 2, 4, 12, or 20 weeks. Cell densities were determined in flatmounted retinas or transverse semithin sections. Expression of glial fibrillary acidic protein (GFAP) was localized on frozen sections or flatmounts by immunofluorescence and confocal microscopy, and GFAP content was evaluated by Western blot analysis. Microglial cells were visualized by binding of isolectin B4 or staining with antibodies to phosphotyrosine residues. The integrity of the blood-retinal barrier was assessed by intravenous injection of Evans blue. RESULTS: The density of Muller cells and microglia was significantly increased at 4 weeks of diabetes compared with nondiabetic controls. GFAP expression in Muller cells was not detected at 4 weeks but was prominent at 12 weeks. The number of astrocytes was significantly reduced at 4 weeks in the peripapillary and far peripheral retina. Shape changes of microglial cells indicated functional activation. Leakage of the blood-retinal barrier was observed at 2 weeks of hyperglycemia, the earliest time point investigated. CONCLUSIONS: The leakage of the blood-retinal barrier before glial reactivity suggests that glia are early targets of vascular hyperpermeability. The individual glial cell types react differentially to the diabetic state. Muller cells undergo hyperplasia preceding GFAP expression, and microglial cells are activated, whereas astrocytes regress. This glial behavior may contribute decisively to the onset and development of neuropathy in the diabetic retina. PMID- 10845625 TI - The lipofuscin fluorophore A2E mediates blue light-induced damage to retinal pigmented epithelial cells. AB - PURPOSE: To determine whether the lipofuscin fluorophore A2E participates in blue light-induced damage to retinal pigmented epithelial (RPE) cells. METHODS: Human RPE cells (ARPE-19) accumulated A2E from 10, 50, and 100 microM concentrations in media, the levels of internalized A2E ranging from less than 5 to 64 ng/10(5) cells, as assayed by quantitative high-performance liquid chromatography (HPLC). Restricted zones (0.5-mm diameter spots) of confluent cultures were subsequently exposed to 480 +/- 20-nm (blue) or 545 +/- 1-nm (green) light for 15 to 60 seconds. Phototoxicity was quantified at various periods after exposure by fluorescence staining of the nuclei of membrane-compromised cells, by TdT-dUTP terminal nick-end labeling (TUNEL) of apoptotic cells and by Annexin V labeling for phosphatidylserine exposure. RESULTS: Nonviable cells were located in blue light- exposed zones of A2E-containing RPE cells, whereas cells situated outside the illuminated areas remained viable. As shown by fluorescence labeling of the nuclei of membrane-damaged cells and by the presence of TUNEL-positive cells, the numbers of nonviable cells increased with exposure duration and as a function of the concentration of A2E used to load the cells before illumination. The numbers of blue light-induced TUNEL-positive cells also increased in advance of the increase in labeling of membrane-compromised cells, a finding that, together with Annexin V labeling, indicates an apoptotic form of cell death. Conversely, blue light- exposed RPE cells that did not contain A2E remained viable. In addition, illumination with green light resulted in the appearance of substantially fewer nonviable cells. CONCLUSIONS: These studies implicate A2E as an initiator of blue light-induced apoptosis of RPE cells. PMID- 10845626 TI - Spatiotemporal expression patterns of 6B4 proteoglycan/phosphacan in the developing rat retina. AB - PURPOSE: To investigate expression of 6B4 proteoglycan/phosphacan, the major constituent of chondroitin sulfate proteoglycan and a possible modulator of neural network formation in the developing central nervous system, in developing rat retina. METHODS: Changes in expression and localization of 6B4 proteoglycan in developing rat retina were investigated by reverse transcription-initiated polymerase chain reaction (RT-PCR), immunohistochemistry, and immunoblot analysis. RESULTS: Semiquantitative RT-PCR revealed that mRNA expression of 6B4 proteoglycan in retinas peaked at postnatal day 14 (P14) and then decreased at P42. Immunohistochemical analyses using MAb 6B4, a monoclonal antibody against 6B4 proteoglycan, revealed faint immunoreactivity in the inner aspects of the retina at embryonal day 16 (E16). At birth, weak immunoreactivity was present in the nerve fiber layer (NFL) and inner plexiform layer (IPL). At P7 and P14, the NFL, IPL, and outer plexiform layer (OPL) stained intensely, but the ganglion cell layer (GCL) remained unstained. Between P21 and P42, immunoreactivity in the NFL and IPL weakened slightly. Immunoblot analyses showed a MAb 6B4 immunopositive band in the retinal soluble fraction treated with chondroitinase ABC. The amount of the immunopositive band increased rapidly as retinal development proceeded. Surprisingly, a significant amount of the immunopositive band was present in the retina even before digestion with chondroitinase ABC, indicating that at least part of 6B4 proteoglycan in rat retina exists in a non proteoglycan form. CONCLUSIONS: The existence of 6B4 proteoglycan/phosphacan was thus demonstrated in rat retina, although some biochemical parameters were different from those of the 6B4 proteoglycan seen in brain. PMID- 10845627 TI - Functional assessment of the regional distribution of disease in a cat model of hereditary retinal degeneration. AB - PURPOSE: To establish a method for the recording of multifocal electroretinograms (MF-ERGs) in animals under fundus control using a scanning-laser ophthalmoscope (SLO) and to analyze the spatial distribution of disease in a strain of Abyssinian cats with a recessively inherited rod-cone degeneration (ARCD). METHODS: Four normal and 12 Abyssinian cats at four different clinical stages of ARCD were examined with the RETIscan MF-ERG system using 61 hexagonal elements within a visual field of approximately 30 degrees radius. The stimulus pattern was generated by the green laser beam (515 nm) of a Heidelberg Engineering HRA SLO, whose power was reduced with a Schott long-pass filter allowing for simultaneous infrared fundus imaging. RESULTS: Topographical recordings could be obtained in all animals except one in stage 4. Amplitudes were minimal at the optic disc and had a slight maximum at the area centralis. Implicit times had a tendency to lower values in the central region, most pronounced in progressed stages of ARCD. The clinical stages of ARCD correlated with a successive generalized loss of amplitude and a rise in implicit time. Without a decrease in retinal illuminance, topographical landmarks like the optic disc were no longer detectable, pointing to stray light as a possible cause. CONCLUSIONS: It was demonstrated that topographical MF-ERG recordings can be obtained in an animal model under fundus control using SLO stimulation. The appearance of retinal landmarks was found to be dependent on sufficient attenuation of laser power. Because the changes in ARCD are more patchy than in human retinitis pigmentosa (RP), a generalized loss of function was detected. However, like in RP, the central area was found to retain a better function than the periphery, especially in later stages of the disease. In summary, fundus controlled methods like the one presented will greatly improve the reliability of MF-ERG in future research on glaucoma, transplantation studies, and evaluation of gene therapy. PMID- 10845628 TI - Effect of instructions on conventional automated perimetry. AB - PURPOSE: To investigate the effect of perimetrists' instructions on automated perimetry thresholds. METHODS: Eighteen volunteers in two age groups participated in a series of three test sessions. Each session consisted of a Humphrey Field Analyzer 30-2 test, a questionnaire, and a customized test program using a Humphrey perimeter to construct frequency of seeing (FOS) curves from which thresholds were calculated, and a descriptive measure of response criterion was derived. The FOS curves were obtained at a central and a peripheral test location within the same test session. The three test sessions differed only by the instructions given. The instructions were adapted from those listed in the manufacturer's instruction manual and were designed to influence participants to respond to the stimuli in a conservative, liberal, or neutral manner. RESULTS: For the 30-2 threshold test, a significant difference in mean deviation was found among the three instruction types (P = 0.001) and between the two age groups (P = 0.001). Although differences were small in the younger subjects (2.04 dB), the means for the responses from liberal to conservative differed by 6.57 dB in the older subjects. Thresholds obtained in a peripheral location by the customized threshold test were found to differ significantly between the age groups (younger group mean, 31.0 dB; older group mean, 27.2 dB) and among the instruction types (liberal mean, 30.9 dB; conservative mean, 28.1 dB; and neutral mean, 30.3 dB; P < 0.001). The descriptive measurement of response criterion suggests that a difference in criteria occurred as a result of the instructions in both peripheral and central locations for both age groups (P = 0.0001). In addition, according to self-reports, liberal instructions caused participants to be more likely to respond, whereas the conservative instructions caused them to be more reluctant to respond. CONCLUSIONS: Perimetrists' instructions can significantly affect obtained automated perimetry thresholds. PMID- 10845629 TI - Molecular spin ladders AB - The structural and magnetic characteristics of the recently discovered quantum spin ladders are summarised and the possibilities of supramolecular chemistry for developing such fascinating materials are discussed. The existing examples of molecular spin ladders clearly show that typical supramolecular and crystal engineering criteria, such as pi-pi overlap, S***S and C-H***S interactions and complementarity of size and shape, are useful in the construction of these magnetic quantum systems with intermediate dimensionalities between one dimensional chains and two-dimensional square lattices. PMID- 10845630 TI - On the electrostatics of cell-membrane recognition: from natural antibiotics to rigid push-pull rods. AB - The question why pore-forming, alpha-helical natural antibiotics are not toxic is discussed within the general context of the interaction of electrostatically asymmetric "nanorods" with neutral, anionic, and polarized bilayer membranes. We suggest that simplification of the structural complexity of natural systems will be necessary to ultimately define the involved subtle balance between constructive and destructive electrostatic interactions. PMID- 10845631 TI - Rigid-rod beta-barrels as lipocalin models: probing confined space by carotenoid encapsulation. AB - Herein, we describe the design, synthesis, structure, and function of synthetic, supramolecular beta-barrel models. Assembly of octi(p-phenylene)s with complementary -Lys-Leu-Lys-NH2 and Glu-Leu-Glu-NH2 side chains yielded water soluble rigid-rod beta-barrels of precise length and with flexible diameter. A hydrophobic interior was evidenced by guest encapsulation. Host-guest complexes with planarized, monomeric beta-carotene within tetrameric rigid-rod beta barrels, and disc micellar astaxanthin J-aggregates surrounded by about dodecameric rigid-rod "bicycle tires" were prepared from mixed micelles by dialytic detergent removal. The significance of these findings for future bioorganic chemistry in confined, intratoroidal space is discussed in comparison with pertinent biological examples. PMID- 10845632 TI - Dynamic analyses on induced-fit gaseous guest binding to organic crystals with a quartz-crystal microbalance AB - The inclusion behavior of gaseous guest molecules in a solid apohost, an orthogonal anthracene-bis(resorcinol)tetraol (1), was investigated with a quartz crystal microbalance (QCM). Compound 1 forms crystals composed of molecular sheets bound together by an extensive hydrogen-bonded network. An apohost of 1 was cast onto a QCM and the binding of gaseous guest molecules was followed as a function of time by observing the decrease in the oscillation frequency, which is directly related to the increase in mass. Ethyl acetate and methyl ethyl ketone were significantly included into the apohost, whereas benzene and cyclohexane were simply adsorbed onto the surface of the solid; all these guests have similar vapor pressures at 25 degrees C. On the other hand, a host analogue 2, a tetramethoxy derivative of 1, barely included these guest molecules. The inclusion amount and the rate of inclusion of ethyl acetate or methyl ethyl ketone showed a drastic increase above a threshold concentration of guests in the gas phase. Thus, the structure of the apohost changed cooperatively in order to bind guest molecules above the threshold guest concentration. This cooperativity of the binding behavior was kinetically analyzed. PMID- 10845633 TI - Synthesis of "sugar-rods" with phytohemagglutinin cross-linking properties by using the palladium-catalyzed Sonogashira reaction AB - A palladium-catalyzed Sonogashira reaction has been applied for the syntheses of divalent "sugar-rods" which exhibited excellent lectin cross-linking properties. The procedure, which involves a tetrakis(triphenylphosphine)palladium-catalyzed cross-coupling reaction between an alkyne and a halogen-bearing sp2-carbon in DMF at 60 degrees C, is very efficient and the dimeric or heterobifunctional "sugar rods" (8-13, 15-17) were isolated in 65-100% yields. Dimers 8a and 15a were both shown to form insoluble cross-linked lattices when mixed with the tetrameric plant lectin from Canavalia ensiformis (Concavalin A, Con A). Moreover, the relative inhibitory properties of the synthetic dimannosides were determined by means of the hemagglutination of rabbit erythrocytes, whereby dimer 15a was shown to be 20-fold more potent than monomeric methyl alpha-D-mannopyranoside. PMID- 10845634 TI - Introduction of heteroatom-based substituents into 1,4-dihydropyridines by means of a halogen-mediated, oxidative protocol: diamination, sulfonylation, sulfinylation, bis-sulfanylation, and halo-phosphonylation processes AB - The natural tendency of 1,4-dihydropyridines to undergo "biomimetic" oxidation to afford pyridinium salts can be switched off and, through the use of reagents that interact electrophilically with the enamine moiety present in the heterocyclic system, it is possible to promote alternative oxidations. In this way, efficient regio- and stereocontrolled synthetic methods have been developed that lead to diversely substituted di- and tetrahydropyridines. These include iodoazidation, diamination, bis-sulfonamidation, sulfonylation, sulfinylation, thiocyanation, sulfanylation, bis-sulfanylation, and halo-phosphonylation processes. PMID- 10845635 TI - Polymer-supported carbene complexes of palladium: well-defined, air-stable, recyclable catalysts for the Heck reaction AB - N-Heterocyclic dicarbene chelate complexes of formula [cis CH2N(H)C=C(H)N(R)C2PdX2] (X = Br, I; R = (CH2)nOH; n = 2, 3) have been prepared and structurally characterized (for X= I, n = 2). The complexes were immobilized on a functionalized polystyrene support (Wang resin) through one of the oxygen centres. The complexes efficiently catalyze the Heck reaction of activated and non-activated arylbromides, are recyclable under aerobic conditions and exhibit hardly any leaching, which is in line with our theoretical investigations on ligand dissociation energies related to Pd0 and PdII centres. PMID- 10845636 TI - Construction of alpha-helix peptides with beta-cyclodextrin and dansyl units and their conformational and molecular sensing properties. AB - In order to apply de novo peptide design to molecular sensing, we designed and synthesized a-helical peptides with beta-cyclodextrin (beta-CDx) as a binding site and a dansyl unit (Dns) as a fluorescence sensing site. The conformational and molecular sensing properties of the peptides with beta-CDx and Dns in various positions were investigated. Circular dichroism and fluorescence measurements revealed that beta-CDx and Dns form intramolecular complexes which depend on their positions in the peptides. In the 17 residual peptides named EK3 and EK3R, in which beta-CDx and Dns were introduced at the fourth and the eighth positions (EK3) or at the eighth and the fourth positions (EK3R), Dns was deeply included in the CDx cavity and formed a more stable self-inclusion complex with CDx than in the peptides EK6 and EK6R, in which these moieties were at the eighth and the fifteenth positions or at the fifteenth and the eighth positions, respectively. The stability of the self-inclusion complex between beta-CDx and Dns controlled the a-helix structure as well as the binding and sensing abilities for the exogenous guests. These results demonstrate the usefulness of peptide tertiary structure for arranging CDx and other functional units, and suggest that this approach is important in the development of a new type of CDx-based sensory system. PMID- 10845637 TI - A density-functional theory based study on the 16O/18O-exchange reactions of the prototype iron-oxygen compounds FeO+ and FeOH+ with H2(18)O in the gas phase AB - The mechanism of the degenerate 16O/18O exchange in the reactions of FeO+ and FeOH+ with water is examined by density functional theory. Based on previous experimental work (Chem. Eur. J. 1999, 5, 1176), two possible reaction pathways are investigated for both systems. The first mechanism consists of one (for FeOH+ + H20) or two (for FeO+ + H20) 1,3-hydrogen migrations from one oxygen atom to the other; the iron atom is not directly involved in these OH bond activations. The second route comprises a series of two (for FeOH+ + H20) or four (for FeO+ + H20) 1,2-hydrogen migration steps which involve the intermediate formations of metal-hydrogen bonds. Both mechanisms are evaluated under consideration of the respective low- and high spin potential-energy surfaces. The computational results show a clear preference for the 1,3-routes occurring on the respective high-spin surfaces bypassing the intermediacy of high-valent iron compounds having FeH bonds. PMID- 10845638 TI - Evaluation of the solid state dipole moment and pyroelectric coefficient of phosphangulene by multipolar modeling of X-ray structure factors AB - The electron density distribution of the molecular pyroelectric material phosphangulene has been studied by multipolar modeling of X-ray diffraction data. The "in-crystal" molecular dipole moment has been evaluated to 4.7 D corresponding to a 42% dipole moment enhancement compared with the dipole moment measured in a chloroform solution. It is substantiated that the estimated standard deviation of the dipole moment is about 0.8 D. The standard uncertainty (s.u.) of the derived dipole moment has been derived by splitting the dataset into three independent data-sets. A novel method for obtaining pyroelectric coefficients has been introduced by combining the derived dipole moment with temperature-dependent measurements of the unit cell volume. The derived pyroelectric coefficient of 3.8(7) x 10-6 Cm 2K-1 is in very good agreement with the measured pyroelectric coefficient of p = 3 +/- 1 x 10-6 Cm-2 K-1. This method for obtaining the pyroelectric coefficient uses information from the X-ray diffraction experiment alone and can be applied to much smaller crystals than traditional methods. PMID- 10845639 TI - Synthesis and reactivity of the Tl1 salts of AB - Synthons Tl1[TCNE]*- (1) and Tl12[TCNE]2- (2), for [TCNE]*- and [TCNE]2-, respectively, in metathesis reactions have been quantitatively prepared and characterized. The structure of 1 was solved and refined in a monoclinic unit cell at 27 degrees C [C2/c, a = 12.6966 (12) angstroms, b=7.7599 (7) angstroms, c=15.5041 (15) angstroms, beta = 96.610 (5) degrees , V= 1517.4 (2) angstroms3, Dcalcd = 2.911 gcm-3, Z=8, R1 = 0.0575, omegaR2=0.0701] and exhibits nuCN absorptions at 2,191 (s) and 2,162 (s) cm-1 consistent with metal-bound [TCNE]*-. The structure of 1 consists of a distorted square antiprismatic octacoordinate Tl1 bound to six monodentate [TCNE]*-s with TlN separations ranging from 2.901 to 3.171 angstroms averaging 3.020 angstroms, and one bidentate [TCNE]*- with TlN separations averaging 3.279 angstroms. The TlN bonding is attributed to electrostatic bonding. The [TCNE]*-s form dimerized zigzag chains with intra- and interdimer separations of 2.87 and 3.29 angstroms, respectively. The tight pi [TCNE](2)2- dimer is diamagnetic and has the shortest intradimer [TCNE]*- distance reported. These synthons for [TCNE]*- and [TCNE]2- in metathesis reactions lead to the precipitation of, for example, TlIX (X = Cl, Br, OAc). Reaction of 1 with MnIII(porphyrin)X (X = Cl, OAc) forms the molecule-based magnets of [MnIII(porphyrin)][TCNE] composition, while the reaction of [CrI(C6H6)2]Br and (Me2N)2CC(NMe2)2Cl2, [TDAE]Cl2, with 1 forms [CrI(C6H6)2] [TCNE] and [TDAE][TCNE]2, respectively. The structure of [TDAE][TCNE]2.MeCN was solved and refined in an orthorhombic unit cell at 21 degrees C [I222, a = 10.2332(15), b = 13.341(6), c = 19.907(8) angstroms, V= 2717.7 angstroms3, Z = 4; Dcalcd = 1.216 gcm-3, R=0.083, Romega = 0.104] and exhibits upsilonCN absorptions at 2,193 (m), 2,174 (s), and 2,163 (s) cm-1 consistent with isolated [TCNE](2)2- , in contrast to the aforementioned TlI bound [TCNE](2)2-. The reaction of 2 with [TDAE]Cl2 forms [TDAE]2+[TCNE]2-. PMID- 10845640 TI - Products from the reaction of meso-tetrakis(4-halophenyl)porphinato-manganese(II) and hexacyanobutadiene (HCBD): formation of pi- AB - The reaction of hexacyanobutadiene (HCBD) and meso-tetrakis(4 chlorophenyl)porphinatomanganese(II) pyridine [MnIITClPPpy] (1Cl) leads to two phases of [MnIIITClPPpy][HCBD].PhMe (alpha-2Clpy, beta-2Clpy). Similarly, the reaction of HCBD and tetrakis(4-bromophenyl)porphinatomanganese(II) pyridine [MnIITBrPPpy] (1Br) leads to two products [MnIIITBrPPpy] [HCBD] * PhMe (2 Brpy) and [MnIIITBrPP][HCBD]*2 PhMe (3Br). The structure of dark-green alpha-2Clpy consists of one porphyrin unit with MnIII in a square pyramidal coordination environment axially bound to one pyridine. The cation forms [MnIIITCIPPpy](2)2+ as cofacial dimerized porphyrins. Each [HCBD]*- is nonplanar with a torsion angle of 170.8(4) degrees about the center C-C bound, and forms [HCBD](2)2- dimers in the solid state with sub-van der Waals contacts of 3.325 and 3.340 angstroms. The magnetic data above 10 K obey the Curie-Weiss expression with a theta of -2.5 K, and mueff (300 K) = 4.91 muB as expected for S=2MnIII and S = 0 [HCBD](2)2-. The magnetic data for alpha-2Clpy can be fit with an zero-field-splitting D of -1.45 K. beta-2 Clpy consists of one porphyrin unit with MnIII in a distorted octahedral coordination environment axially bound to py and to a monodentate [HCBD]*- bound via an exo-nitrile. [HCBD]*- is nonplanar with a torsion angle of 169.7(5) degrees about the center C-C bound. The ueff (350 K) is 5.09 muB; however, the magnetic data do not obey the Curie-Weiss expression above 70 K. The low temperature data may be fit with a theta of -5.4 K. The data was modeled to an isolated S = 2 and S = 1/2 dinuclear spin system with J/kappaB = - 90 K. Decomposition of [HCBD]*- to [C4(CN)5O]- was evidenced by the determination of the structure of [MnIIITCIPP][C4(CN)5O] 2PhMe (3ClO). Crystals of 3 Cl-O were prepared by reaction of HCBD and 1Cl in the presence of a drop of water. The molecular structure consists of [HCBD]*-trans-mu-N-2,3-bound to [MnIIITBrPP]+ forming a 1-D coordination polymer of alternating [MnIIITBrPP]+ and [HCBD]*-. The intrachain Mn***Mn distance was 10.675(3) angstroms, with important interchain Mn***Mn distances of 10.832, 11.016, and 14.696 A. The magnetic data were fit to a Curie-Weiss law (10 < T< 290 K) with a theta of -3.5 K, and D = 0.3 K with mueff = 4.97 muB at 300 K. PMID- 10845641 TI - Optically and redox-active ferroceneacetylene polymers and oligomers AB - The palladium-catalyzed Sonogashira reaction can be used to build optically active, oligomeric 1,2,3-substituted ferrocenes up to the tetramer, as well as polymers, by sequential coupling of optically active (ee > 98 %), planar chiral iodoferroceneacetylenes and ferroceneacetylenes. (SFC)-1-Iodoferrocene-2 carbaldehyde (1) was reduced to the alcohol and methylated to give the corresponding methyl ether, which was Sonogashira-coupled with HC(triple bond)CSiEt3, resulting in (RFc)-1-(C(triple bond)CSiEt3)-2-methoxymethylferrocene (4) (79%, three steps). Orthometalation with tBuLi followed by quenching with 1,2 diodoethane gave (RFc)-1-(C(triple bond)CSiEt3)-2-methoxymethyl-3-iodoferrocene (5). Deprotection of the acetylene with nBu4NF resulted in (RFc)-1-ethynyl-2 methoxymethyl-3-iodoferrocene (6), which was Sonogashira-coupled with itself to produce an optically active polymer. Deprotection of 4 with nBu4NF and Sonogashira coupling of the product with 5 resulted in the dinuclear ferrocene 9. Deprotection of 9 and coupling with 5, followed by deprotection of the resulting acetylene 11, gave the trinuclear ferrocene 12. Another such sequence involving 11 and 5 produced a tetranuclear ferrocene 13. To study the electronic communication in such oligomers in more detail, two symmetrical, closely interrelated, trinuclear ferrocenes 18 and 19 were synthesized. The redox potentials of all the ferrocenes and the ferroceneacetylene polymer were determined by cyclic and square-wave voltammetry. All the metallocenes were investigated by UV/Vis spectroscopy. A linear relationship was found between lambdamax and l/n (n=number of ferrocene units in the oligomer). The polymer displayed two redox waves in the cyclic voltammogram, at 0.65 and 0.795 V. The corresponding mixed-valence oligoferrocene cations were synthesized from four ferroceneacetylenes, and their metal-metal charge transfer bands were examined by UV/Vis-NIR. The resonance exchange integrals Had, calculated on the basis of spectral information from the metal - metal charge transfer (MMCT) bands, were between 290 and 552 cm-1. PMID- 10845642 TI - Palladium/phosphite catalyst systems for efficient cross coupling of aryl bromides and chlorides with phenylboronic acid AB - The Suzuki reaction of aryl bromides is efficiently catalyzed by palladium/ phosphite complexes generated in situ. The influence of ligand, base, and different additives is examined. The process tolerates various functional groups and catalyst turnover numbers up to 820,000 are obtained even with deactivated aryl bromides. For the first time it is shown that palladium/phosphite complexes also catalyze efficiently the Suzuki reaction of aryl chlorides. PMID- 10845643 TI - Synthesis and structural characterization of hexa-tert-butyl-hexa-peri hexabenzocoronene, its radical cation salt and its tricarbonylchromium complex AB - The hexa-tert-butyl substituted hexa-peri-hexabenzocoronene was synthesized in an overall yield of 83% from 4-tert-butylphenylacetylene. The key step was the oxidative cyclodehydrogenation of hexa(4-tert-butylphenyl)benzene with anhydrous FeCl3 in CH2Cl2 The high solubility of hexa-tert-butyl-hexa-peri hexabenzocoronene in common organic solvents allowed a comprehensive spectroscopic characterization of this compound in solution. Electrochemical oxidation at -30 degrees C in the presence of tetrabutylammonium hexafluoroarsenate led to the formation of a stable radical cation salt. Reaction of hexa-tert-butyl-hexa-peri-hexabenzocoronene with an excess of tricarbonyl(naphthalene)chromium in THF/dioxane afforded a mixture of mono- and bis-tricarbonylchromium complexes which could be separated by chromatography. The molecular structures of the parent compound, its radical cation salt and its mono tricarbonylchromium complex were determined by X-ray analysis and discussed in detail. Remarkably, the crystal structures of these compounds are mainly dominated by the formation of dimers of the aromatic cores. PMID- 10845644 TI - Catalytic asymmetric hydroboration/amination and alkylamination with rhodium complexes of 1,1'-(2-diarylphosphino-1-naphthyl)isoquinoline AB - Catecholboronate esters formed by asymmetric hydroboration of arylalkenes are not directly converted to amines by reaction with hydroxylamine-O-sulfonic acid. Prior conversion to a trialkylborane by reaction with ZnEt2 or MeMgCl permits a subsequent amination reaction to occur with essentially complete retention of configuration, leading to a range of primary alpha-arylalkylamines in up to 97% enantiomeric excess (ee). Secondary, but not tertiary amines may be formed by a related pathway when in situ generated alkylchloramines are employed as the aminating agent. The catalytic asymmetric hydroboration, beta-alkylation and amination steps may be combined in a single stage. Overall, this provides a practical procedure for the synthesis of enantiomerically enriched arylamines, exemplified inter alia by the synthesis of (S)-1,2,3,4-tetrahydro-1-naphthylamine in 95-97% ee and of (R)-N-(cyclohexyl)-1'-(4-methoxyphenyl)ethylamine in 93% ee. PMID- 10845645 TI - Cationic ruthenium allenylidene complexes as catalysts for ring closing olefin metathesis AB - A series of well accessible cationic ruthenium allenylidene complexes of the general type [(eta6-arene)(R3P)RuCl(=C=CR'2)]+ X- is described which constitute a new class of pre-catalysts for ring closing olefin metathesis reactions (RCM) and provide an unprecedented example for the involvement of metal allenylidenes in catalysis. They effect the cyclization of various functionalized dienes and enynes with good to excellent yields and show a great tolerance towards an array of functional groups. Systematic variations of their basic structural motif have provided insights into the essential parameters responsible for catalytic activity which can be enhanced further by addition of Lewis or Bronsted acids, by irradiation with UV light, or by the adequate choice of the "non-coordinating" counterion X-. The latter turned out to play a particularly important role in determining the rate and selectivity of the reaction. A similarly pronounced influence is exerted by remote substituents on the allenylidene residue which indicates that this ligand (or a ligand derived thereof) may remain attached to the metal throughout the catalytic process. X-ray crystal structures of the catalytically active allenylidene complexes 3b.PF6 and 15.OTf as well as of the chelate complex 10 required for the preparation of the latter catalyst are reported. PMID- 10845646 TI - 17O NMR spectroscopy of sulfolenes (2,5-dihydrothiophene-1,1-dioxides) and sultines (3,6-dihydro-1,2-oxathiin-2-oxides)--experiment and quantum calculations: synthesis of 4,9-dioxo-1,2-oxathiacyclodecane-2-oxide, a new heterocycle AB - The products of hetero-Diels-Alder reactions (sultines) and cheletropic addition reactions (sulfolenes) between 1,3-dienes and sulfur dioxide can be distinguished by their 17O NMR shifts. Experimental data have been collected for derivatives of 3,6-dihydro-1,2-oxathiin-2-oxide and of 2,5-dihydrothiophene-1,1-dioxide. This data was then compared with that calculated by the gauge independent atomic orbital (GIAO) method at the HF/6-31 + G(d,p) and HF/6-311 + G(3df, 2p) levels of theory with geometries optimized by MP2/6-31G(d) calculations. GIAO-MBPT(2) calculations were also performed with the 6-31 + G(d,p) basis set. The adduct between (E)-1-methoxybutadiene and SO2 is sulfolene 3, the ozonolysis of which in SO2 followed by work-up with ethanol provided (2RS,3SR,6SR)-(31), (2RS,3RS,6SR) (32), and (2RS,3RS, 6RS)-2,6-diethoxy-3-methoxy-1,4-oxathiane-4,4-dioxide (33). Single-crystal X-ray diffraction studies are reported for 32 and 33. Ozonolysis of the hetero-Diels-Alder adduct of SO2 with 1,2-dimethylidenecyclohexane produced 4,9-dioxo-1,2-oxathiacyclodecane-2-oxide (34), the first member of a new class of sulfur heterocycles. PMID- 10845647 TI - Dimerization of formic acid--an example of a "noncovalent" reaction mechanism AB - The pulse deposition technique allows selectively the isolation of monomeric or dimeric formic acid in argon matrices at 7 K. Warming of matrices containing the monomer M from 7 K to 40 K results in the decrease of M and formation of a dimer B. This dimer is also labile, and further warming finally produces a second dimer A. By comparison with density functional theory (DFT) calculations and gas phase IR spectra taken from the literature, the latter dimer A was identified as the C2h-symmetrical cyclic dimer. The unstable dimer B was identified as the acyclic Cs-symmetrical dimer. An activation energy of 2.3 kcal mol(-1) was calculated for the B --> A rearrangement at the B3LYP/ 6-311 ++ G(d,p) level of theory, which is in qualitative agreement with the experimental finding of a slow thermal reaction under the conditions of matrix isolation. PMID- 10845648 TI - Catalytic methanol oxidation over copper: observation of reaction-induced nanoscale restructuring by means of in situ time-resolved X-ray absorption spectroscopy AB - The catalytically active copper phase for the partial oxidation of methanol is studied by means of time-resolved extended X-ray absorption fine structure (EXAFS) spectroscopy combined with the detection of the catalytic turnover. It is found that the active form of the copper is a strained nanocrystalline form of the metal. The metal is no longer made up from large crystallites but contains a defect structure in which oxygen is already intercalated. PMID- 10845649 TI - Species of Caligus Muller, 1785 (Copepoda: Caligidae) parasitic on marine fishes of Taiwan. AB - Six species of copepods of the genus Caligus (Caligidae, Siphonostomatoida) parasitic on marine fishes of Taiwan are reported. They are: C. absens n. sp. from Priacanthus blochii and P. macracanthus; C. epinepheli Yamaguti, 1936 from Scolopsis vosmeri; C. kanagurta Pillai, 1961 from Decrapterts kurroides; C. laticaudus Shiino, 1960 from Lutjanus vitta and Parapristiopoma trilineatum; C. nengai Rangnekar, Rangnekar & Murti, 1953 from Triacanthus biaculeatus; and C. rotundigenitalis Yu, 1933 from Drepane punctata, Liza macrolepis and Terapon jarbua. C. distortus Pillai & Natarajan, 1977 is relegated to a synonym of C. nengai and C. multispinosus Shen, 1957 reported by Lin et al. (1994) from the cultured sea bream Acanthopagrus schlegeli is a misidentification for C. rotundigenitalis Yu, 1933. PMID- 10845650 TI - A new species of Aponurus Looss, 1907 (Digenea: Lecithasteridae) in butterflyfishes (Perciformes: Chaetodontidae) from New Caledonia. AB - Aponurus chelebesoi n. sp. is described from Chaetodon auriga, C. citrinellus, C. ephippium, C. flavirostris, C. lineolatus, C. melannotus, C. mertensii, C. pelewensis, C. lunulatus, C. vagabundus, Coradion altivelis, Forcipiger flavissimus, Heniochus acminatus, H. chrysostomus and H. monoceros from the southern coast of New Caledonia. It is distinguished from most species in the genera Aponurus (synonym Brachadena) and Lecithophyllum by its claviform (as opposed to oval to subglobular) vitelline lobes. Three species, A. pyriformis, Lecithophyllum vogeae and Brachadena cheilonis, have similar claviform vitelline lobes, but differ from A. chelebesoi in their tandem testes and the distinct egg size. PMID- 10845651 TI - Criconematidae (Nematoda) from oak forests in two nature reserves in Russia. AB - Seven criconematid species: Ogma menzeli, Criconema annuliferum, C. princeps, Criconemoides pleriannulatus, Mesocriconema solivagum, M. xenoplax and Xenocriconemella macrodora were recovered from soil samples collected from oak forests in the Shulgan-Tash and Voronezh nature reserves in Russia. Examination of C. pleriannulatus by SEM confirmed the absence of submedian lobes and revealed the presence of a small lip annule; juveniles exhibit fringe-like cuticular ornamentation of the annules. Ogma menzeli and C. pleriannulatus are new geographical records for Russia, and all species have already been reported in association with different oak species with the exception of M. solivagum and C. pleriannulatus. Criconemoides mutabilis Eroshenko, 1980 is proposed as ajunior synonym of C. pleriannulatus Ebsary, 1979. PMID- 10845652 TI - Steganoderma szidati n. sp. (Trematoda: Zoogonidae) from Galaxias maculatus (Jenyns) and G. platei Steindachner in Patagonia, Argentina. AB - Steganoderma szidati n. sp. is described from the intestine of two freshwater fishes, Galaxias maculatus and G. platei (Galaxiidae), from Andean lakes in Patagonia, Argentina. This freshwater zoogonid species fits in the genus Steganoderma (sensu Bray, 1987) because of the length of the caeca and the position of the vitelline follicles. The new species is characterised by possessing 6-13 vitelline follicles situated between the anterior margin of the ventral sucker and the posterior margin of the testis. The gonads are in the anterior hindbody and the ovary is anterior to the right testis. The cirrus has two conspicuous spines at its distal end, and the seminal vesicle always exhibits a constriction. The excretory vesicle never reaches the level of the posterior margin of the testes. PMID- 10845653 TI - Surface morphology of Probolocoryphe uca (Sarkisian, 1957) (Digenea: Microphallidae) from Kuwait bay. AB - The surface ultrastructure of Probolocoryphe uca (Digenea: Microphallidae), recovered from a rat experimentally fed on crabs, Nanosesarma minutum (Brachyura: Grapsidae), naturally infected with the metacercariae, was studied by scanning electron microscopy. The flukes were leaf-like, ventrally concave and pyriform or ovoid in outline. The anterior end was modified into a sucker-like organ, comprising a protrusible disc-shaped structure surrounded by single-pointed spines. This organ is probably involved in the attachment and feeding process in a manner similar to the action of the oral suckers. Apart from the sucker-like organ, the entire tegumental surface was covered with triangular spines with multi-pointed tips. Ciliated, dome-shaped papillae were observed, singly or in groups, arranged symmetrically on the sucker-like organ and around the oral and ventral suckers. Kuwait Bay constitutes a new geographical record and the crab N. minutum is a new second intermediate host record for P. uca. PMID- 10845654 TI - Observations on the morphology and life-cycle of Procerovum varium (Onji & Nishio, 1916) (Trematoda: Heterophyidae). AB - The life-cycle of Procerovum varium (Digenea: Haplorchiinae) was studied experimentally and the morphology of stages in the life-cycle has been described and illustrated. Infections with adult flukes were found in the pond heron Ardeola grayii and heavy infections with metacercariae were found, attached to the liver of the fish Oryzias melastigma (Oryziatidae) occurring in a freshwater stream situated in Visakhapatnam, India. The cercariae developing in the snail Thiara tuberculata possessed typical haplorchiine features and were characterised by the presence of numerous cystogenous glands. Early stages of metacercarial development occurred free in the muscles of the fish intermediate host. The larvae reached the liver at 5 days post-infection, encystment commenced 2 days later and 15-day-old metacercariae were found to be infective to chicks, ducks and mice that served as suitable experimental hosts. The adult flukes obtained from natural and experimental infections showed many intraspecific variations, especially in the size and shape of the expulsor which depend on the quantity of sperm it contains. The validity of various species described in the genus and differentiated on the basis of differences in the size of the expulsor has been examined. It is concluded that only three species of the genus, namely P. varium, P. cheni and P. calderoni, are valid. "P. sisonli" of Chen (1949) is confirmed as a synonym of P. varium. P. varium is reported for the first time from India. PMID- 10845655 TI - Wardium canarisi n. sp. (Cestoda: Hymenolepididae) parasite of Arenaria melanocephala (Aves: Charadrii) of Alaska. AB - The authors describe and illustrate Wardium canarisi n. sp. (Cestoda: Hymenolepididae), an intestinal parasite of Arenaria melanocephala (Aves: Charadrii) from Alaska, characterised by a strobila 20-40 mm long, 10 aploparaksoid hooks 19-21 microm long, a short cylindrical cirrus (40 microm) covered with very minute spines (0.2 microm), and a short (8-18 microm) and extremely narrow (1 microm) copulatory vagina. These characters have no equivalent, even approximate, among the 27 species of Wardium parasitic in the Charadrii which are reviewed. The genus Debrosia Spassky, 1987 appears to be justified. PMID- 10845656 TI - Disturbed bile secretion and cytochrome P450 function during the acute state of experimental colitis in rats. AB - BACKGROUND/AIMS: A variety of hepatobiliary abnormalities has been described in patients with inflammatory bowel diseases. However, the pathophysiological mechanisms leading to these liver alterations are poorly understood. The aim of the present study was to investigate parameters of liver function in a trinitrobenzenesulfonic acid (TNB)-induced rat colitis model. METHODS: Glucose output, bile acid secretion, bile acid uptake, and the cytochrome P-450 metabolic capacity during TNB-colitis were studied in the perfused liver model. Furthermore, hepatic bile acid- and glycogen content was measured. To evaluate the inflammatory response in the colon and liver, NF-kappaB/Rel was quantified by electrophoretic mobility shift assays. As an NF-kappaB/Rel regulated gene the inducible NO-synthase (NOS2) was evaluated by Western blot analysis. As possible mediators released from the inflamed colon into the portal vein, endotoxin and the stable metabolite of prostaglandin I2 (6-keto-prostaglandin-F1alpha) were determined. RESULTS: Glucose output, bile acid secretion, bile acid uptake, and cytochrome P-450 metabolic capacity decreased on the first and second day of TNB colitis. Hepatic bile acid content increased at day 14 of colitis. Glycogen content was reduced, most likely due to an inadequate chow intake of these animals. A low level of portal endotoxin was detectable during the first 2 days of colitis. In addition, 6-keto-prostaglandin-F1alpha was clearly increased in portal blood. NF-kappaB/Rel binding activity and inducible NOS2 were strongly positive in the colon during colitis. Although low levels of portal endotoxin were measured during the first 2 days of colitis, no significant NF-kappaB/Rel activity and NOS2 induction were detected in the liver. CONCLUSION: Our results indicate that during the acute state of the TNB-colitis, bile acid secretion and cytochrome P-450 function are disturbed in the absence of distinct inflammatory changes in the liver. PMID- 10845657 TI - Hepatocyte-supported serum-free culture of rat liver sinusoidal endothelial cells. AB - BACKGROUND/AIMS: A major problem in rat liver endothelial cell culture is the rapid loss of cells after 48 h. This study aimed to develop a protocol that allowed easy maintenance and proliferation of sinusoidal endothelial cells in serum-free culture for 5-6 days. METHODS: Cells isolated from adult rat liver by collagenase digestion were purified by centrifugal elutriation and cultured on glutaraldehyde-crosslinked collagen. RESULTS: At high plating densities cells could be maintained serum-free for 6 days in the presence of hydrocortisone and basic fibroblast growth factor; at lower plating densities medium had to be supplemented with additional growth-promoting factors. Conditioned medium of adult rat hepatocytes proved to be the most effective growth stimulus; it increased thymidine incorporation, DNA content and cell number per dish with a half-maximal effect at 20% (v/v). Cell proliferation was also observed with either vascular endothelial growth factor, phorbol ester or conditioned media from FAO or HEPG2 liver cell lines provided the cultures were additionally supplemented with 1% newborn calf serum. Vascular endothelial growth factor was detected in all conditioned media. In the absence of hepatocyte-conditioned medium, 1% serum helped to maintain cultures; it itself exerted a low proliferative effect. Higher serum concentrations (>5%), however, led to cell loss after 48 h. The numerous sieve plates of 6-h-old cells progressively disappeared during culture and were replaced by randomly distributed pores, which later grouped together at cell-cell borders. More than 90% of the cells endocytosed acetylated low-density lipoprotein. CONCLUSIONS: The study shows that cultured hepatocytes secrete growth-promoting substances that stimulate in vitro endothelial cell proliferation in the absence of serum; this effect could be mimicked by the combined addition of vascular endothelial growth factor and 1% serum. The new media formulations should facilitate future research on liver endothelial cells in mono- or coculture. PMID- 10845658 TI - Determination of non-transferrin-bound iron in genetic hemochromatosis using a new HPLC-based method. AB - BACKGROUND/AIMS: Non-transferrin-bound iron may play a major pathogenic role in iron overload diseases due to its high hepatic uptake and potential damaging effect. The aim of this study was to evaluate the relevance of measuring serum non-transferrin-bound iron levels in genetic hemochromatosis using a new high performance liquid chromatography-based method. METHODS: This method includes a presaturation step of transferrin with cobalt(II) in order to avoid secondary deplacement of non-transferrin-bound iron toward transferrin during the assay. Six genetic hemochromatotic patients were followed serially during venesection treatment. RESULTS/CONCLUSIONS: The results indicate: (i) that this new method permits detection of non-transferrin-bound iron when transferrin is not fully saturated, (ii) that non-transferrin-bound iron levels persist almost until the completion of treatment, (iii) that non-transferrin-bound iron levels are well correlated with transferrin saturation for a given patient, and (iv) that despite some individual variations, a transferrin saturation value lower than 35% usually corresponds to the disappearance of non-transferrin-bound iron. PMID- 10845659 TI - Clinical significance of anti-neutrophil cytoplasmic antibodies (ANCA) in autoimmune liver diseases. AB - BACKGROUND/AIMS: The clinical relevance of anti-neutrophil cytoplasmic antibodies (ANCA) in autoimmune liver disease is unclear. Defining the antigenic specificities of ANCA in these diseases may improve their clinical significance. METHODS: We studied the target antigens of ANCA in 88 patients with autoimmune hepatitis, 53 patients with primary biliary cirrhosis, and 55 patients with primary sclerosing cholangitis by indirect immunofluorescence, antigen-specific enzyme-linked immunosorbent assays, and immunodetection on Western blot, using an extract of whole neutrophils as a substrate. We related the data to clinical symptoms of autoimmune liver disease. RESULTS: By indirect immunofluorescence, ANCA were present in 74% of patients with autoimmune hepatitis, 26% of patients with primary biliary cirrhosis, and 60% of patients with primary sclerosing cholangitis. Major antigens were catalase, alpha-enolase, and lactoferrin. The presence of ANCA as detected by indirect immunofluorescence was associated with the occurrence of relapses in autoimmune hepatitis, with decreased liver synthesis function in primary biliary cirrhosis and in primary sclerosing cholangitis, and with increased cholestasis in primary sclerosing cholangitis. ANCA of defined specificities had only limited clinical relevance. CONCLUSIONS: ANCA as detected by indirect immunofluorescence seem associated with a more severe course of autoimmune liver disease. The target antigens for ANCA in these diseases include catalase, alpha-enolase, and lactoferrin. Assessment of the antigenic specificities of ANCA in autoimmune liver disease does not significantly contribute to their clinical significance. PMID- 10845660 TI - Increased intestinal permeability to macromolecules and endotoxemia in patients with chronic alcohol abuse in different stages of alcohol-induced liver disease. AB - BACKGROUND/AIMS: No information is yet available about the influence of alcohol abuse on the translocation of larger molecules (Mr>1200) through the intestinal mucosa in man. The present study aimed to determine the intestinal permeability to macromolecules in patients with chronic alcohol abuse and mild to more advanced stages of liver disease, and to measure the concentration of endotoxins in the plasma, as these compounds derive from the intestinal flora and are suspected to contribute to the development of alcoholic liver disease (ALD). METHODS: The permeability to polyethylene glycol Mr 400, Mr 1500, Mr 4000, and Mr 10,000 and endotoxin plasma concentrations were measured in 54 patients with alcoholic liver disease, 19 of them with cirrhosis, and in 30 non-alcoholic healthy controls. RESULTS: Permeability to polyethylene glycol Mr 400 was found to be unchanged in patients with ALD in comparison to healthy controls, whereas polyethylene glycol Mr 1500 and Mr 4000 were recovered in about twice as high concentrations in the urine of ALD patients (p<0.01). Polyethylene glycol Mr 10,000 was detected significantly less frequently in urine from healthy controls (0/30) than in urine of patients with alcoholic liver disease (20/54, p<0.01). Endotoxin concentrations in the plasma of alcoholics were increased more than 5 fold compared to healthy controls (p<0.01). CONCLUSIONS: The results of this study indicate that alcohol abuse impairs the function of the intestinal barrier, which might enhance the translocation of bacterial toxins, thereby contributing to inflammatory processes in alcoholic liver disease. PMID- 10845661 TI - Screening of subclinical hepatic encephalopathy. AB - BACKGROUND/AIMS: Subclinical hepatic encephalopathy adversely affects daily functioning. The aim of this study was to determine which elements of daily life have predictive value for subclinical hepatic encephalopathy. METHODS: The study was performed in 179 outpatients with liver cirrhosis. Subclinical hepatic encephalopathy was diagnosed using psychometric tests with normal values corrected for age (Number Connection Test A and the Digit Symbol Test) and automated analysis of the electroencephalogram (EEG). Daily functioning was measured with the Sickness Impact Profile (SIP), a quality of life questionnaire, containing 136 statements. Patients with and without SHE were compared for differences in response to all statements by univariate analysis, and subsequently by multivariate analysis of potential discriminating statements. RESULTS: SHE was diagnosed in 48 patients (27%). Thirty-six statements were significantly more often true for patients with subclinical hepatic encephalopathy. Multivariate analysis showed that five statements of the SIP, related to alertness, sleep and rest, fine motor skills and work, have independent predictive power for subclinical hepatic encephalopathy. CONCLUSION: Combining these statements predictive for subclinical hepatic encephalopathy with patient characteristics enables physicians to assess the probability of subclinical hepatic encephalopathy in the individual cirrhotic patient at the bedside or in the outpatient clinic. PMID- 10845662 TI - Increased expression of connective tissue growth factor in fibrotic human liver and in activated hepatic stellate cells. AB - BACKGROUND/AIMS: Connective tissue growth factor is a recently described mitogenic protein implicated in a variety of fibrotic disorders. Connective tissue growth factor may be a downstream mediator of the pro-fibrotic and mitogenic actions of transforming growth factor-beta, promoting extracellular matrix deposition and fibrogenesis. As transforming growth factor-beta is considered important to the pathogenesis of hepatic fibrosis, we examined the possible contribution of connective tissue growth factor to this process. METHODS: Connective tissue growth factor expression was examined in normal and fibrotic human and rat livers using RT-PCR and ribonuclease protection assays, and in primary cultures of rat hepatic stellate cells by Northern and Western blotting. RESULTS: Ribonuclease protection assays demonstrated connective tissue growth factor mRNA was increased 3-5-fold in human fibrotic liver compared with normal. RT-PCR showed this mRNA was increased in carbon-tetrachloride-treated rat liver. Northern analysis showed connective tissue growth factor mRNA was increasingly expressed during progressive activation of cultured rat hepatic stellate cells. Western analysis confirmed that freshly isolated hepatic stellate cells secreted relatively little connective tissue growth factor compared with hepatic stellate cells activated in culture. Hepatic stellate cells stimulated with transforming growth factor-beta showed increased expression of connective tissue growth factor mRNA and protein. CONCLUSIONS: Connective tissue growth factor mRNA is consistently upregulated in human liver cirrhosis of various aetiologies, supporting a role for this growth factor in hepatic fibrogenesis. Our studies suggest that hepatic stellate cells may be an important source of hepatic connective tissue growth factor in vivo, particularly following stimulation with transforming growth factor-beta. PMID- 10845663 TI - A p160ROCK-specific inhibitor, Y-27632, attenuates rat hepatic stellate cell growth. AB - BACKGROUND/AIMS: p160ROCK, a serine/threonine protein kinase, is a direct RhoA target mediating RhoA-induced assembly of focal adhesions and stress fibers. Recently, Rho signaling pathways were reported to play an important role in the activation of rat hepatic stellate cells (HSC). The aim of this study was to investigate the mechanism of action of a p160ROCK-specific inhibitor, Y-27632, on cultured rat HSC. METHODS: HSC were isolated from normal rat livers and cultured on fibronectin-coated dishes. The cell morphology and actin cytoskeleton were studied with phase contrast and fluorescence microscopy, respectively. Immunoblot analysis was used to examine phosphorylation of focal adhesion kinase and extracellular signal-regulated kinase, and the expression of cell cycle associated proteins. HSC proliferation was measured by quantitating the percentage of cells that exhibited nuclear incorporation of 5-bromodeoxyuridine. Type I collagen gene expression and accumulation in HSC culture media were evaluated by Northern blot and enzyme-linked immunosorbent assay, respectively. RESULTS: Y-27632 consistently blocked cell spreading and suppressed RhoA-induced formation of stress fibers in HSC. In addition, Y-27632 inhibited phosphorylation of focal adhesion kinase and extracellular signal-regulated kinase. Cells treated with Y-27632 failed to proliferate, in contrast to untreated spread cells. This shape-dependent block in cell proliferation correlated with a failure to increase cyclin D1 protein level and to down-regulate the cell cycle inhibitor p27. Y 27632 decreased type I collagen gene expression and accumulation in HSC culture media. CONCLUSIONS: Our findings indicate that p160ROCK-mediated actin stress fiber assembly is involved in the pathophysiology of hepatic fibrogenesis and suggest that inhibitors of the RhoA-ROCK pathway might be useful therapeutically in liver fibrogenesis. PMID- 10845664 TI - Induction of cell proliferation arrest and apoptosis in hepatoma cells through adenoviral-mediated transfer of p53 gene. AB - BACKGROUND/AIM: Loss of p53 function is common in hepatocellular carcinoma and is associated with an extremely poor prognosis. The aim of the study was to evaluate the biologic effect of adenoviral-mediated gene transfer of wild-type p53 gene in four hepatoma cell lines with different p53 genetic makeup. METHODS: Recombinant adenovirus expressing wild-type p53 was used. Recombinant adenoviruses with either an empty expression cassette or expressing beta-galactosidase gene served as controls. RESULTS: High-level expression of wild-type p53 was achieved with adenoviral-mediated gene transfer. The expressed p53 protein showed nuclear localization and its expression was associated with an induction of p21 and bax expression. Expression of the p53 gene was associated with inhibition of tumor cell proliferation and induction of apoptosis. Expression of p53 was also associated with an upregulation of CD95 (Apo-1/Fas) gene expression, which may predispose the tumor cells to undergo apoptosis induced by the Fas Ligand/Fas cytolytic pathway. An additional anti-tumor effect, in terms of allowing the replication-defective adenovirus to replicate, was observed in hepatoma cells with homozygous deletion of p53 genes and to a lesser extent, hepatoma cells with mutated p53 genes. CONCLUSIONS: These data showed that adenoviral-mediated gene transfer is effective in delivering p53 gene to tumor cells, and the multiple pathways involved in their antitumor activities. PMID- 10845665 TI - Complement activation induced by ischemia-reperfusion in humans: a study in patients undergoing partial hepatectomy. AB - BACKGROUND/AIM: Activation of the complement system is induced by ischemia reperfusion (I/R) in animal models. Whether I/R also induces complement activation in humans is not known. Here, we investigated complement activation in patients undergoing major liver resection. METHODS: In 11 of 17 patients, the hepatoduodenal ligament was clamped, making the liver transiently ischemic (HEMI+; mean ischemia time, 42 +/- 18 min); 6 patients were operated without clamping (HEMI-). Activation at plasma level (circulating activation products) was studied in blood samples collected prior to surgery and 5, 24 and 48 h thereafter. Parameters analyzed were C4b/c and C3b/c, C4d and C3d, C3a, as well as complexes between complement and C-reactive protein (CRP), which reflect CRP induced complement activation. Activation at tissue level (C3 and C4 fixation) was studied in liver biopsies obtained before and after resection. RESULTS: In plasma, post-operative levels of C4b/c and C3b/c were not different from baseline levels in both groups. Mean plasma levels of C4b/c and C3b/c were significantly decreased at 24 h post-surgery in the HEMI+ group (p=0.02 and p=0.07). At the same time, levels of C4d-CRP and C3d-CRP were significantly increased (p<0.01 for both parameters). At tissue level, activated complement fragments were observed intracellularly in some pericentral hepatocytes. In I/R livers, large numbers of hepatocytes were positively stained for all complement activation products. CONCLUSIONS: Our data show that in situ complement activation via the classical route occurred during liver resection and that ischemia and/or reperfusion may have contributed to activation. Levels of complement activation products in the circulation were low, showing that transient ischemia had no severe influence on systemic complement activation, suggesting a locally contained response. PMID- 10845666 TI - Positron emission tomography scanning in the evaluation of hepatocellular carcinoma. AB - BACKGROUND/AIMS: 18F-fluorodeoxyglucose uptake allows estimation of glucose metabolism by tumor cells using positron emission tomography (PET). We evaluated the role of PET imaging in the diagnosis of hepatocellular carcinoma. METHODS: PET images were collected after intravenous injection of 8-12 mCi of 18F-FDG in 20 patients with hepatocellular carcinoma (HCC). PET tumor activity level was assessed on a scale of 1 to 4 compared to normal liver tissue. The PET score was compared with abdominal computerized tomography (CT) scan results and between tumors of different grades and differentiation. RESULTS: Of the 20 patients studied, 11 (55%) had positive PET scans (PET score: 3 or 4) while nine (45%) were negative (PET score: 1 or 2). CT scan was positive in 18 patients (90%) and negative in two (10%). PET, however, revealed metastases in three patients that were not seen on CT. On pathological review, well-differentiated and low-grade tumors had lower PET scores. Comparison of the well-differentiated with the moderately- and poorly-differentiated tumors revealed a statistically significant difference. No statistical significance was observed between the moderately- and poorly-differentiated tumors or between different tumor grades and PET scores. CONCLUSIONS: The sensitivity of PET in diagnosis of HCC was 55% compared to 90% for CT scanning, although only PET detected some tumors (including distant metastases). Well-differentiated and low tumor grades had lower activity on PET and correspondingly lower PET scores. PET imaging may help assess tumor differentiation and may be useful in the diagnosis and staging and prognostication of HCC as an adjunct to CT. PMID- 10845667 TI - Nitric oxide release and enhancement of lipid peroxidation in regenerating rat liver. AB - BACKGROUND/AIMS: Clarification of the role of lipid peroxidation in the onset of liver proliferation has been hampered by the fact that both higher and lower lipid peroxidation have been reported after two-thirds partial hepatectomy. Recently, it has been shown that nitric oxide might be involved in the control of early responses after partial hepatectomy. We analysed the possible involvement of nitric oxide production in lipid peroxidation levels during liver regeneration. METHODS: Sham-operated, hepatectomised and sham and hepatectomised rats pretreated with two inhibitors of oxide nitric synthesis (aminoguanidine or N(G)-monomethyl-L-arginine) were used throughout. Animals were killed at 1, 3, 5 and 15 h after surgery. Cytosolic superoxide dismutase and microsomal-lysosomal catalase activities were measured. Lipid peroxidation levels were measured as thiobarbituric acid-reactive substances and conjugated dienes. Cytosolic nitrate (a stable metabolic product of nitric oxide) was enzymatically determined. Inducible-type nitric oxide synthase (iNOS) was analysed in hepatic cytosol by immunoblotting. DNA synthesis 24 and 48 h after surgery was assessed by [3H]thymidine incorporation. RESULTS: Increased lipid peroxidation was found in total homogenate, cytosol and microsomes. The hepatic cytosolic content of nitrates increased, reaching the highest values at 5 h posthepatectomy. Aminoguanidine or N(G)-monomethyl-L-arginine pretreatment blocked the rise of nitric oxide production and lipid peroxidation levels and decreased the DNA synthesis. The increase in hepatic iNOS protein expression at 5 h after partial hepatectomy disappeared with aminoguanidine pretreatment. CONCLUSIONS: Our experiments suggest that nitric oxide plays a role in the proliferation mechanism, although it is responsible, at least in part, for the enhanced lipid peroxidation. PMID- 10845668 TI - Increased incidence of HFE C282Y mutations in patients with iron overload and hepatocellular carcinoma developed in non-cirrhotic liver. AB - BACKGROUND/AIMS: Histological and biochemical iron overload has been reported in non-tumoral liver of most patients presenting an hepatocellular carcinoma (HCC) developed in non-cirrhotic liver (NCL). The aim of our study was to investigate HFE mutations in patients with HCC in NCL. METHODS: Thirty-five patients with HCC in NCL were included either retrospectively or prospectively. Clinical data, iron and viral status, and HFE gene mutations were compared between groups with (I+, n = 19) or without histological iron overload (I-, n = 16). RESULTS: Twenty per cent of patients were HBV or HCV positive. Fifty-four per cent had hepatocytic iron overload at histology. Mean hepatic iron concentration was 100.2 +/- 14.6 micromol/g in I+ versus 23.2 +/- 2.1 micromol/g in I- (p<0.001). Among the 19 I+ patients, eight mutations were found: two C282Y/C282Y, three C282Y/WT, two C282Y/H63D and one H63D/H63D. None of these mutations was found in the I- group. There was no significant difference concerning the H63D heterozygous mutation between I+ or I- patients. CONCLUSIONS: In patients with HCC in NCL, HBV and HCV markers are rare (20%), and mild iron overload is frequent (54%). In patients with HCC in NCL and iron overload, C282Y mutations are frequent (36.8% of cases) and significantly increased (p<0.009) compared to HCC in NCL without iron overload; these mutations are mostly heterozygous. H63D heterozygosity is not associated with liver iron overload. Because of the small size of the series, HFE C282Y mutation should be investigated on a larger scale in patients with HCC in NCL with iron overload in order to confirm this association. PMID- 10845669 TI - Enhancement of immune tolerance via induction of NK1.1 positive liver-associated lymphocytes under immunosuppressive conditions. AB - BACKGROUND/AIMS: The liver was previously shown to play a critical role in oral tolerance induction. A subset of liver-associated-lymphocytes expressing NK1.1 marker (NK1.1+ LAL) have killing activities and it has been suggested that they play a role in immune modulation. FK506 is a powerful immunosuppressive agent affecting T-cell differentiation and function. The exact pathway involved in peripheral tolerance induction using this drug remains unknown. The aim of the present study was to determine the interaction between FK506 and NK1.1+ LAL in induction of peripheral immune tolerance in the experimental colitis model. METHODS: Colitis was induced in C57 mice by intracolonic instillation of trinitrobenzenesulfonic acid (TNBS). Mice received five oral doses of colonic proteins extracted from TNBS-colitis colonic wall with and without FK506 treatment. The effect of FK506 treatment on NK1.1+ LAL was tested by cell-sorting and cytotoxicity assay. Colitis was assessed by standard clinical, macroscopic and histologic scores. RESULTS: Both FK506 treatment and oral tolerance induced a significant increase in NK1.1+ LAL number and cytotoxicity function. FK506 treatment enhanced the effect of oral tolerance on amelioration of disease activity. Orally tolerized mice treated with FK506 had no mortality nor increase in body weight, and manifested significant improvement in disease macroscopic and microscopic scores. CONCLUSIONS: This study shows for the first time that immune tolerance induced by both oral administration of an antigen and by FK506 treatment may be mediated via enhancement of NK1.1+ LAL. This subset of lymphocytes may play an immunoregulatory role in immune tolerance induction. PMID- 10845670 TI - Anti-ICAM-1 blockade reduces postsinusoidal WBC adherence following cold ischemia and reperfusion, but does not improve early graft function in rat liver transplantation. AB - BACKGROUND/AIM: The present in vivo study investigated the impact of a monoclonal antibody directed against the intercellular adhesion molecule-1 (ICAM-1) on initial microvascular reperfusion injury after liver transplantation. METHODS: Orthotopic, syngeneic liver transplantation including arterial reconstruction was performed in male Lewis rats after 24 h graft storage in University of Wisconsin (UW) solution at 4 degrees C. Animals received either an anti-ICAM-1 antibody (n=7), an IgG1 control antibody (n=8) or saline only (n=7). Hepatic microvascular alterations during the initial 90 min of reperfusion were assessed using intravital fluorescence microscopy. Early graft dysfunction was determined by analysis of bile flow. RESULTS: After treatment with anti-ICAM-1 mAb, hepatic microvascular perfusion was found improved when compared with that of IgG1- and saline-treated controls. In addition, anti-ICAM-1 mAb effectively reduced the number of permanently adherent white blood cells in postsinusoidal venules (284.4+/-59.1 mm(-2) vs IgG1: 371.9+/-26.7 mm(-2) and saline: 431.8+/-46.4 mm( 2); p<0.05). In contrast, the number of stagnant white blood cells in sinusoids was higher (p<0.05) in liver grafts with blocked ICAM-1 (320.6+/-17.2 mm(-2)) compared with that of IgG1- (215.2+/-11.1 mm(-2)) and saline-treated controls (226.4+/-14.0 mm(-2)). Measurement of hepatic uptake of fluorescent-labeled latex particles did not reveal significant differences in phagocytic activity. Finally, bile flow also did not differ between the three groups studied. CONCLUSION: Together these results indicate that ICAM-1 is involved in the process that mediates white blood cells adherence in postsinusoidal venules, whereas in hepatic sinusoids other mechanisms apart from ICAM-1-mediated white blood cells adherence seem to be fundamental for posttransplant white blood cells accumulation. Our data further suggest that white blood cells adherence in postsinusoidal venules via ICAM-1 does not make a major contribution to the pathogenesis of early cold ischemia/reperfusion injury after liver transplantation. PMID- 10845671 TI - Olone modulates the therapeutic effect of interferon to eliminate preferentially the hepatitis B virus precore stop mutant. AB - BACKGROUND/AIMS: The aim of this study was to understand the changes in the proportion of hepatitis B virus precore stop mutant during the course of prednisolone primed interferon (IFN) therapy. METHODS: Three groups of patients were included: patients receiving prednisolone-primed IFN treatment (Group I, n=31), IFN treatment only (Group II, n=29), and placebo (Group III, n=25). The proportion of precore stop mutant was measured by a quantitative amplification created restriction site method. RESULTS: Distinct patterns of the progression of the proportion of mutant were found among these three groups. A steady increase in the proportion of mutant was observed only in Group III patients. In Group II patients, the presence of a higher percentage of mutant (> 25%) immediately before IFN treatment was predictive for the subsequent clearance of hepatitis B e antigen (HBeAg) (p<0.01), but not for complete anti-viral response (p>0.05). Prednisolone pretreatment resulted in an increase in the proportion of mutant in patients with initially low percentages (< or = 25%) of mutant. During the period of IFN treatment, both the relative and absolute amount of the precore stop mutant decreased significantly in Group I patients who cleared HBeAg. The presence of such a decrease in this group of patients was predictive for both HBeAg clearance and complete anti-viral response. CONCLUSIONS: Our data suggest that prednisolone serves as a modulator to enhance elimination of precore stop mutant by IFN, which advocates the benefit of corticosteroid pretreatment in an area where the precore mutants are prevalent. PMID- 10845672 TI - Human leukocyte antigen DRB1 1302 protects against bile duct damage and portal lymphocyte infiltration in patients with chronic hepatitis C. AB - BACKGROUND/AIMS: To confirm the immune reaction of hosts in chronic hepatitis C, we examined the association of human leukocyte antigen (HLA) DR with the histopathological outcome including bile duct damage and steatosis, which are characteristic of hepatitis C virus (HCV) infection. METHODS: One hundred and fifty-five patients with chronic HCV infection were examined. The pathological appearance of liver biopsy specimens was evaluated by both Knodell's histological activity index and examination of bile duct damage and steatosis. HLA DRB1 was determined by the polymerase chain reaction sequence-specific oligonucleotide probe method. RESULTS: HLA DRB1 1302 was found with significantly higher frequency in patients without than with bile duct damage (34.8% vs. 4.7%, p=0.0001, p corrected by Bonferroni's inequality method=0.002). It was also found more frequently in patients without marked portal lymphocyte infiltration (28.6% vs. 7.7%, p=0.0015, p corrected by Bonferroni's method=0.03). HLA DRB1 1101 was found more frequently in patients without than with piecemeal necrosis (p=0.004). In contrast, the frequency of HLA DRB1 1502 tended to be higher in patients with than without piecemeal necrosis and marked portal lymphocyte infiltration (p=0.015 and p=0.03, respectively). HLA DRB1 1201 and 0802 were seen more frequently in bile duct damage-negative (p=0.02) and piecemeal necrosis-negative patients (p=0.03), respectively. Interestingly, serum HCV levels of HLA DRB1 1302 positive patients were significantly higher than those of 1302-negative patients (mean: 7.7 Meq/ml vs. 3.1 Meq/ml, p=0.0007). CONCLUSION: These findings suggest that some histopathological changes in chronically HCV-infected livers could be caused by the host's immune reaction regulated by HLA DR. PMID- 10845673 TI - Utility of early testing for HCV viremia as predictive factor of sustained response during interferon or interferon plus ribavirin treatment. AB - BACKGROUND/AIM: To evaluate the utility of early testing for hepatitis C viremia as a predictor of treatment outcome during interferon or combination therapy. METHODS: We studied 184 patients with chronic hepatitis C who received interferon and were monitored for HCV RNA. Sixty-two patients received interferon alone for 12 months and 122 patients, who were still HCV RNA positive at 2 months, received an additional 12-month course of interferon and ribavirin combination therapy. RESULTS: Using this strategy, sustained response occurred in a total of 34 patients (18.5%). Independent variables associated with sustained response were HCV genotype (p=0.06), viral load < or = 5.1 logs/ml (p= 0.005) and negative HCV RNA at 1 month (p<0.0001) in the interferon group, and female sex (p=0.04), genotype (p=0.03), viral load < or = 5.5 logs/ml (p=0.01), normal ALT (p=0.001) and decline in viral load > or = 1.2 logs/ml after 2 months of interferon monotherapy (p<0.001) and negative viremia at 5 months of ribavirin onset (p<0.0001) in the combination therapy group. Persistence of viremia at 1 month of interferon monotherapy and at 5 months of combination therapy were the strongest predictors of non-response (negative predictive value of 100% and 99%, respectively). CONCLUSIONS: Qualitative assessment of HCV RNA during treatment is the strongest predictor of sustained response during interferon or combination therapy for chronic hepatitis C. PMID- 10845674 TI - Decrease of wild-type and precore mutant duck hepatitis B virus replication during lamivudine treatment in white Pekin ducks infected with the viruses. AB - BACKGROUND/AIMS: Lamivudine, an antiviral agent, has been used in the treatment of chronic hepatitis B, but little is known about its effect on intrahepatic replication of hepatitis B virus. We investigated the effect of lamivudine on the replication of wild-type and precore mutant duck hepatitis B virus (DHBV) in the liver and serum of DHBV carrier ducks. METHODS: Chronic carrier ducks with either wild-type or precore mutant DHBV were treated for 2 weeks with either low-dose (20 mg/kg, p.o., b.i.d.) or high-dose lamivudine (100 mg/kg, p.o., b.i.d.) or were untreated. Serum levels of DHBV DNA were examined serially by slot-blot hybridization. A second group of chronic carrier ducks was treated for 12 weeks with lamivudine (100 mg/kg, p.o., b.i.d.) or was untreated. The amount of DHBV DNA in serum and its various replicative intermediates in the liver were serially examined by slot-, Southern, and Northern blot methods. RESULTS: In the 2-week treatment study, concentration of DHBV DNA in serum treated with low- and high dose lamivudine was reduced to 10.8% and 1.1% of the control level in wild-type DHBV carriers, and to 2.3% and 0.48% in precore mutant DHBV carriers, respectively. In the 12-week treatment study, concentration of DHBV DNA in serum at the end of treatment was reduced to <0.65% and <5.36% in wild-type and precore mutant DHBV carriers, respectively. Southern and Northern blot analyses revealed that the various replicative forms of DHBV DNA in the liver were decreased in all treated ducks, but, covalently closed circular DNA and RNA intermediates tended to remain unchanged. CONCLUSIONS: Our results showed that lamivudine could reduce both wild-type and precore mutant DHBV levels in the liver through inhibition of the reverse transcription step, but complete elimination of the viruses from liver is difficult even by relatively long-term lamivudine monotherapy, suggesting a need for some additional therapy to obtain complete clearance. PMID- 10845675 TI - "What's in a name?" Improving the care of cirrhotics. PMID- 10845676 TI - Iron, friend or foe? "Freedom" makes the difference. PMID- 10845677 TI - Portal vein thrombosis in adults: pathophysiology, pathogenesis and management. PMID- 10845678 TI - Images in hepatology. Periportal lymph edema in a patient with acute hepatitis A. PMID- 10845679 TI - Triggering of acute alcoholic hepatitis by alpha-interferon therapy? PMID- 10845680 TI - Fatal fulminant hepatic failure associated with benzbromarone. PMID- 10845681 TI - The formation and mechanism of multimerization in a freeze-dried peptide. AB - Atrial natriuretic peptide (ANP) is a peptide with 25 amino acid residues (hANP 4 28) and one intra-chain disulfide bond. We used the size-exclusion chromatography (SEC), sodium dodecyl sulfate in polyacrylamide gel electrophoresis (SDS-PAGE), and reversed-phase liquid chromatography with electrospray mass spectrometry (LC MS) methods to examine the freeze-dried products of ANP to determine the types, sizes, and amounts of the multimer formation in different stability protocols (by varying the conditions with lyophilization cycles, excipients, storage temperatures, and times). Under the non-annealing lyophilization cycle or lyophilization with high concentration of bulking agent (mannitol), multimer formation increased with increasing storage times. Two kinds of multimers were observed; the major portion is reducible and the minor portion is non-reducible. The reducible multimers are disulfide-linked multimers as determined by LC-MS. The non-reducible multimer was mainly a dimer, possibly linked by a covalent bond between the side chain of tyrosine in one molecule and the dehydroalanine intermediate in another molecule, based on the evidences of the mass of the non reducible dimer along with the elution position in SEC, and the change of the UV spectrum in the aromatic region. The analysis of degradants suggests that the mechanism start from an beta-elimination of disulfide linkage to form a free thiolate ion (HS-) and a dehydroalanine-type peptide intermediate. The HS- then catalyzed ANP to form the disulfide-linked multimers. The dehydroalanine-type ANP intermediate then reacted with another ANP molecule to form a non-disulfide linked dimer through reaction with the side chain of tyrosine. These results suggest that the source of multimer formation be initiated by phase transition (from amorphous to crystalline phase) either in the freeze-dried process or during storage. That phase change may induce a drastic change in pH and moisture to damage the peptide. The detailed mechanism and the kinetics of ANP multimerization are discussed. The formation of the multimers was diminished by using the thermal treatment (the annealing step) with a proper ratio of mannitol to ANP peptide in the lyophilization, and/or increase of the acetate buffer concentration in the formulation. PMID- 10845682 TI - The enhancing effect of soybean-derived sterylglucoside and beta-sitosterol beta D-glucoside on nasal absorption in rabbits. AB - The aim of this study was to elucidate the efficiency of soybean-derived sterylglucoside (SG) and its main component beta-sitosterol beta-D-glucoside (Sit G), as nasal absorption enhancers. Nasal administration of verapamil with SG and Sit-G showed the higher bioavailabilities (60.4 and 90.7%, respectively) than that with lactose (39.8%). It was clear that SG and Sit-G promoted the absorption of verapamil through nasal mucosa. To elucidate the mechanism, we measured the calcein leakage from liposomes by incubation with SG, Sit-G, oleic acid, soybean derived sterol, and beta-sitosterol to investigate transcellular absorption and measured the changes in intracellular Ca2+ concentrations ([Ca2+]i) by Sit-G to analyze paracellular absorption. The large amount of calcein leakage induced by enhancers was consistent with an enhancement of bioavailability of verapamil and insulin following nasal administration (oleic acid < SG < Sit-G). Moreover, Sit-G increased [Ca2+]i in the medium containing Ca2+, but not in Ca2+ free medium. This result suggested that Sit-G increases the fluidity of the mucosal membrane and facilitates Ca2+ influx from extracellular sources. In conclusion, a possible explanation for SG and Sit-G to promote drug absorption, is that they may affect both paracellular pathway and transcellular pathways caused by pertubation of lipid. PMID- 10845683 TI - Novel therapeutic nano-particles (lipocores): trapping poorly water soluble compounds. AB - The development of stable spherical lipid-coated drug particles that are termed 'lipocores' is reported here. Unlike conventional lipid-based particles (i.e. liposomes, emulsions, micelles), these particles are comprised solely of a core of a poorly water soluble drug surrounded by polyethyleneglycol conjugated lipid (PEG-lipid) and are formed via a 'kinetic' trapping process. These lipocore particles were made with the acyl chain of 16 carbon length (C16) acyl-chain derivatives of paclitaxel or vinblastine and with the polyene antifungal hamycin. Formation of the particles occurred regardless of the type of PEG-phospholipid used (i.e. acyl chain length, chain saturation, and polymer length) and could also be formed with the negatively charged lipid N-glutaryl-dioleoyl phosphatidylethanolamine (DOPE-GA). Images from both freeze-fracture electron microscopy and electron cryo-microscopy revealed solid spherical structures with no internal lamellae for the PEG-lipid particles made with the C16 derivatives of paclitaxel (BrC16-T) or vinblastine (C16-Vin). From a solute distribution study of lipocores made with BrC16-T and distearoyl-phosphatidylethanolamine-PEG2000 (DSPE-PEG2000), the particles were found to have no measurable aqueous captured volume. Fluorescence anisotropy and order parameter measurements revealed the core material of these particles to be highly immobilized. The mole ratio of BrC16-T:lipid in the lipocores was typically > 90: < 10 and as high as 98:2, and the refrigerated lipocores were stable for several months. BrC16-T/DSPE-PEG2000 lipocores of 50-100 nm particle size were far less toxic than paclitaxel (Taxol) after intraperitoneally (i.p.) or intravenously (i.v.) administration in mice and were active against i.p. and subcutaneously (s.c.) planted human (OvCar3) ovarian carcinoma grown in SCID mice. It is believed the high drug:lipid ratio, the stability, and therapeutic efficacy of these novel particles make them a paradigm for delivery of poorly water soluble drugs and/or their hydrophobic derivatives. PMID- 10845684 TI - Development of a more rapid, reduced serum culture system for Caco-2 monolayers and application to the biopharmaceutics classification system. AB - The objectives were: (1) to develop a more rapid, reduced serum culture system for Caco-2 monolayers, relative to the traditional 21-day, 10% fetal bovine serum (FBS) system; and (2) to determine the biopharmaceutical drug classification of an oral therapeutic agent using this new system. Caco-2 cells were grown in the six well format on polycarbonate filters, in medium containing 2% iron supplemented calf serum (sCS) and a combination of growth factors and hormones. After 4 days in culture, permeabilities of three marker compounds (metoprolol, mannitol, and taurocholate) across monolayers were determined, and compared to permeabilities from the traditional 21-day, 10% FBS system, using cells at similar passage number. Cell morphology, degree of cell differentiation, and the presence of two efflux pumps were assessed. The 2% sCS model was also used to classify the permeability of an oral therapeutic agent as high or low. No difference in permeability was observed for metoprolol transport (P = 0.38) between the two culture methods, and the values obtained were independent of passage number of the cells. Mannitol permeability was about 2-fold higher from the 2% sCS system, as compared to the 10% FBS system. Taurocholate permeability was low indicating the 2% sCS culture at 4 days lacked this particular active transporter capability. Electron micrographs of cells grown in the 2% sCS system at 4 days revealed the presence of microvilli and tight junctions. P-glycoprotein and an efflux pump for furosemide were functionally present. The 2% sCS system indicated the oral therapeutic agent as highly permeable, which agreed with the 10% FBS system. This new system provides a rapid, accurate, and economical option for passive permeability determination, and appears to be applicable to the proposed Biopharmaceutics Classification System (BCS). PMID- 10845685 TI - Improved bioavailability of vitamin E with a self emulsifying formulation. AB - A single dose study was conducted to evaluate the bioavailability of a novel self emulsifying vitamin E preparation, in comparison with that of a commercial product, Natopherol, available as soft gelatin capsules under fasted condition. The self-emulsifying preparation achieved a faster rate and higher extent of absorption. A statistically significant difference was observed between the values of the two preparations in the parameters AUC, Cmax and Tmax. Moreover, the 90% confidence interval of the logarithmic transformed AUC values of the self emulsifying preparation over those of the soft gelatin capsule product was found to be between 2.1 and 4.1, suggesting an increase in bioavailability of between 210 and 410%. As for Cmax, the 90% confidence interval was between 2.1 and 3.0. However, no statistically significant difference was observed between the t(1/2) values estimated from the plasma concentration versus time data of the two preparations. The values are also comparable to those reported in the literature. PMID- 10845686 TI - Macromolecular prodrugs. VIII. Synthesis of polymer-gemfibrozil conjugates. AB - Gemfibrozil is covalently linked to two similar polymers: poly[alpha,beta-(N-2 hydroxyethyl-DL-aspartamide)] and poly[alpha,beta-(N-3-hydroxypropyl-DL aspartamide)]. The synthesised polymer drug conjugates differ in average molecular mass, type of covalent bonding, length of spacer, drug-loading and solubility. PMID- 10845687 TI - The mechanical properties of compacts of microcrystalline cellulose and silicified microcrystalline cellulose. AB - The mechanical properties of compacts of unlubricated microcrystalline cellulose and silicified microcrystalline cellulose were evaluated using the diametric tensile test. The results suggested that, under comparable testing conditions, compacts of silicified microcrystalline cellulose exhibited greater strength than those of microcrystalline cellulose. In addition to enhanced strength, silicified microcrystalline cellulose compacts exhibited greater stiffness and required considerably more energy for tensile failure to occur. Comparison of the data with that obtained for a dry blend of silicon dioxide/microcrystalline cellulose suggested that the functionality benefits of silicification were not due to a simple composite material model. PMID- 10845688 TI - Liposomes encapsulating polymeric chitosan based vesicles--a vesicle in vesicle system for drug delivery. AB - Drug delivery systems comprising vesicles prepared from one amphiphile encapsulating vesicles prepared from a second amphiphile have not been prepared previously due to a tendency of the bilayer components of the different vesicles to mix during preparation. Recently we have developed polymeric vesicles using the new polymer-palmitoyl glycol chitosan and cholesterol in a 2:1 weight ratio. These polymeric vesicles have now been encapsulated within egg phosphatidylcholine (egg PC), cholesterol (2:1 weight ratio) liposomes yielding a vesicle in vesicle system. The vesicle in vesicle system was visualised by freeze fracture electron microscopy. The mixing of the different bilayer components was studied by monitoring the excimer fluorescence of pyrene-labelled polymeric vesicles after their encapsulation within egg PC liposomes or hexadecyl diglycerol ether niosomes. A minimum degree of lipid mixing was observed with the polymeric vesicle-egg PC liposome system when compared to the polymeric vesicle hexadecyl diglycerol ether niosome system. The polymeric vesicle-egg PC vesicle in vesicle system was shown to retard the release of encapsulated solutes. 28% of 5(6)-carboxyfluorescein (CF) encapsulated in the polymeric vesicle compartment of the vesicle in vesicle system was released after 4 h compared to the release of 62% of encapsulated CF from plain polymeric vesicles within the same time period. PMID- 10845689 TI - The effect of spacers on the delivery of metered dose aerosols of nedocromil sodium and disodium cromoglycate. AB - The effect of the spacers (Fisonair, Breath-A-Tech, Volumatic and Nebuhaler) on the in vitro aerosol characteristics of two propellant-driven metered dose inhalers (MDIs), Tilade (nedocromil sodium) and Intal (disodium cromoglycate), was studied. The measurement was carried out on a Marple-Miller impactor operating at 30 l/min. Five actuations were collected for the drug assay. The results showed that Tilade (label claim 2 mg active per actuation) and Intal (label claim 5 mg active per actuation) generated aerosols with a fine particle mass (FPM, i.e. mass of particles 5 microm in the aerosol) of 0.34 mg (S.D. 0.01, n = 4) and 0.02 mg (S.D. 0.01, n = 4) per actuation, respectively. For both inhalers, large volume spacers increased (Fisonair > Nebuhaler > Volumatic) while small volume spacer (Breath-A-Tech) decreased the FPM. The FPM (per actuation) for Tilade with Fisonair, Nebuhaler, Volumatic and Breath-A-Tech was 0.52 (0.03), 0.45 (0.03), 0.41 (0.04) and 0.09 (0.04) mg, respectively, while for Intal the corresponding values were 0.41 (0.02), 0.32 (0.04), 0.28 (0.03) and 0.08 (0.01) mg. Thus, the fine particle mass can be either increased or decreased, depending on the spacer selected. In addition, all spacers significantly reduced the coarse particle (> or = 10 microm) mass, with Fisonair, Breath-A-Tech, Nebuhaler and Volumatic producing only 7.6, 0.4, 5.2 and 2.6, respectively of that from Tilade alone and 15.6, 0.7, 5.4 and 4.1%, respectively of that from Intal alone. The general trends for Tilade and Intal were similar but not quantitatively identical. The proper choice of spacers is therefore important for the optimal delivery of Tilade and Intal. PMID- 10845690 TI - The influence of carrier morphology on drug delivery by dry powder inhalers. AB - Alpha-lactose monohydrate was prepared to have different morphological features but with similar particle size. The crystal shape and surface smoothness of lactose were quantified by a number of shape descriptors and these were supported qualitatively by the visual examination of scanning electron (SE) micrographs of the crystals. All batches of lactose were subjected to a similar history of processing before blending separately with micronised salbutamol sulphate (SS) in a ratio of 67.5:1, w/w, using similar procedures. In vitro deposition of SS from these formulations was investigated after aerosolisation of the formulations at 60 l min(-1) via the Rotahaler and the Cyclohaler into a twin stage liquid impinger. The formulations prepared using the different batches of lactose produced different deposition profiles of SS. The fine particle (< 6.4 microm) fraction (FPF) of aerosolised SS varied from 12.6 +/- 2.4 to 25.6 +/- 1.5% after aerosolisation from the Cyclohaler whilst it changed from 15.0 +/- 2.2 to 24.4 +/ 0.8% after aerosolisation from the Rotahaler. The fine particle dose (FPD) and dispersibility of SS followed a similar trend to the change in the FPF of the drug. No significant difference (ANOVA P > 0.05) was observed for the deposition profiles of SS after aerosolisation from the Rotahaler and the Cyclohaler. The FPF and dispersibility of SS increased with either the surface smoothness (P < 0.01) or elongation ratio (P < 0.01) of lactose crystals. The t-ratio values of FPF and dispersibility of SS generated by changes in the surface smoothness were similar to those resulting from changes in elongation ratio. Increasing either the surface smoothness or the elongation ratio of lactose crystals will increase the potentially respirable fraction of SS from dry powder formulations for inhalation. PMID- 10845691 TI - Transdermal iontophoretic delivery of salmon calcitonin. AB - Electrically enhanced transdermal delivery of salmon calcitonin could be useful for chronic treatment of postmenopausal osteoporosis and other clinical indications as a superior alternative to parenteral delivery. Calcitonin (50 microg/ml) formulation was prepared in citrate buffer (pH 4.0). Epidermis separated from human cadaver skin was used. Most iontophoresis studies were done at a current density of 0.5 m A cm2. Silver/silver-chloride electrodes were used and calcitonin was found to be best delivered under the anode. The relationship between calcitonin flux and current density during iontophoresis was linear. Passive flux was zero. Flux increased with increasing concentration up to 250 microg/ml but then it levels off. Thus, transdermal delivery of salmon calcitonin may be accomplished to achieve therapeutic levels. PMID- 10845692 TI - Release of mifepristone from biodegradable matrices: experimental and theoretical evaluations. AB - Diffusion of mifepristone in poly [(D,L) lactide-co-glycolide)] films was studied by release experiments. Five 50/50 copolymers of increasing molecular weights were used. The degradation effects were shown by gel permeation chromatography (GPC). Release kinetics show the effect of copolymer molecular weights on diffusion and degradation properties of loaded films. A new theoretical model for drug release from a biodegradable matrix was proposed with two assumptions: correlation of the diffusion coefficient with the polymer molecular weight and existence of a first order degradation kinetic. Higuchi's equation is verified at early time and the diffusion coefficient in the non-degraded polymer can be measured. The degradation constant is determined at long time and is compared with the results of GPC. PMID- 10845693 TI - A study of the adhesive-skin interface: correlation between adhesion and passage of a drug. AB - The phenomena taking place at the patch-skin interface, in particular the adhesion of a pressure sensitive adhesive (PSA) film loaded with drug and the partition of the drug between the patch and the skin were correlated. The kinetics of adhesion as well as those of drug passage were studied in detail. Adhesion data were collected from peel test either on skin or on a polymer model. Passage of the drug was studied in a simple system composed of PSA film stuck on skin. In some experiments the film was left on the skin throughout the experiment; in others, it was periodically removed and stuck on again to keep the adhesion force constant during the whole of the experiment. We observed a rapid increase of the drug content in the skin until a plateau was reached. One adhesion for the whole experiment or several adhesions gave a similar curve. The main difference was the rate of increase of skin drug content and the value of the plateau. Different hypotheses concerning the relationship between the adhesion of this PSA and changes in the flux of drug have been put forward. However, it is difficult to extrapolate from this model to the in vivo situation because of variation both between individuals and with time. PMID- 10845694 TI - Influence of cyclodextrin complexation on the in vitro permeation and skin metabolism of dexamethasone. AB - The influence of complexation of a model drug, dexamethasone acetate (DMA), with beta-cyclodextrin (beta-CyD) and hydroxypropyl-beta-cyclodextrin (HP-beta-CyD) on the in vitro permeation through hairless mouse skin and on skin metabolism have been investigated. Complexation with CyDs increased the amount of DMA permeated in the order of 2.0 and 3.0 times for beta-CyD and HP-beta-CyD, respectively. The partition coefficient, between stratum corneum and buffer (K(SC/buffer)), for DMA decreased when the drug was an inclusion complex, being greatest for DMA/HP-beta CyD complex. Complexation protected the drug against skin metabolism. The increase of skin permeation and stability of the model drug in the skin suggest that the complexation with beta-CyD and HP-beta-CyD is a rational way to improve the physical-chemical properties of drugs for use in transdermal delivery systems. PMID- 10845695 TI - Parameters influencing the antigen release from spray-dried poly(DL-lactide) microparticles. AB - Microparticles were produced by spray-drying from a high molecular weight polylactide (PLA R207) for the development of long-lasting controlled release systems of vaccines, which may be designed to obviate the need for booster doses. The current investigation considered the effect of both technological parameters (inlet air temperature and spray rate of feed) and polymeric solutions (polymer concentration and nature of organic solvents) on characteristics of microparticles (morphology, size and antigen loading) containing a water-soluble model antigen (bovine serum albumin, BSA). Following parameters chosen, microparticles were characterized by a mean size from 3.08 +/- 0.06 to 9.43 +/- 0.26 microm and a BSA loading from 2.45 +/- 0.13 to 18.20 +/- 2.25% (w/w). The BSA release rate from microparticles varied from 11.17 +/- 2.20 to 92.60 +/- 3.46% in 24 h. The modification of the inlet temperature, the spray-rate of feed or the use of a mixture of dichloromethane/chloroform (DCM/CFM) instead of DCM alone resulted in the modification of the BSA burst release. This burst release was followed by a BSA release rate slower for microparticles with a low BSA loading. Moreover, the increase of the R207 concentration resulted in a decrease of the BSA release rate while the burst release was not modified. SDS-PAGE electrophoresis and isoelectric focusing analyses of the BSA released from microparticles confirmed the preservation of its physicochemical characteristics. Together, results showed that the spray-dried microparticles loaded with hydrophilic antigen could be used as a potential delivery system for the long lasting controlled release of vaccines. PMID- 10845696 TI - Role of the tertiary and quaternary structures in the stability of dimeric copper, zinc superoxide dismutases. AB - The equilibrium unfolding process of human Cu,Zn superoxide dismutase has been quantitatively monitored through circular dichroism and fluorescence spectroscopy as a function of increasing guanidinium hydrochloride concentration. The process occurs through the formation of a monomeric intermediate species following a three-state transition equilibrium. Comparison with the stability of the prokaryotic Cu,Zn SOD from P. leiognathi shows that the eukaryotic enzyme is more stable than the prokaryotic enzyme by approximately 3 kcal/mol. This difference is due to the monomer-to-unfolded equilibrium, while the dimer-to-monomer equilibrium is comparable for the two enzymes despite their different intersubunit interactions. These results are confirmed by the unfolding of the copper-depleted derivatives. The Cu,Zn superoxide dismutase represents a good example of how evolution has found two independent quaternary assemblies maintaining the same dimer stability. PMID- 10845697 TI - Solution structure of an N-capping peptide from the N-terminal leucine-repeat region of hepatitis delta antigen. AB - Hepatitis delta virus (HDV) is a satellite virus of the hepatitis B virus (HBV) which provides the surface antigen for the viral coat. The RNA genome of HDV encodes two proteins, the small delta antigen and the large delta antigen, which differ only with the latter having an additional 19 amino acids at the C terminus. Previously, we have shown that dAg24-50, a synthetic peptide corresponding to residues 24-50 of the N-terminal leucine-repeat region of hepatitis delta antigen, binds to the viral RNA and forms an alpha-helical conformation in TFE-containing solution. However, it exhibited low alpha-helicity (less than 5%) in the absence of TFE. In order to obtain biologically active delta antigen peptides with higher structural stability in solution, an N-capping 21-residue polypeptide corresponding to residues 24-38 of hepatitis delta antigen (dAg(Cap24-38am)) was synthesized and, surprisingly, its solution structure was found to be a stable alpha-helix (64%) by circular dichroism and 1H NMR techniques. Moreover, the structure of the capping box shows the characteristic L shaped bend perpendicular to the helix axis. This structural knowledge provides a molecular basis for understanding the role of the N-terminal leucine-repeat region of hepatitis delta antigen and has a significant potential for the development of diagnostic and therapeutic methods for HDV. PMID- 10845698 TI - Role of Ser-65 in the activity of alpha-galactosidase A: characterization of a point mutation (S65T) detected in a patient with Fabry disease. AB - Fabry disease is a genetic disorder caused by deficient activity of alpha galactosidase A (alpha-Gal A). Recent gene analysis of a Fabry patient revealed a point mutation (S65T) resulting in a significant decrease of enzyme activity (Chen, C.-H., et al. (1998) Hum. Mutat. 11, 328-330). In order to evaluate the role of Ser-65 in the alpha-Gal A activity and the molecular mechanism of its deficient enzyme activity in mammalian cells, we prepared gene products of S65T, S65A, and E66D mutations of alpha-Gal A by using an expression system with baculovirus/insect cells and characterized the kinetic and physical properties of those purified enzymes. The Km values of mutant enzymes were 3.5 (S65T), 3.4 (S65A), and 2.3 mM (E66D), using 4-methylumbelliferyl alpha-D-galactoside as a substrate, and the Vmax values were 2.7 x 10(6) (S65T), 3.4 x 10(6) (S65A), and 2.5 x 10(6) units/mg (E66D), respectively, which were similar to those of the normal enzyme (Km, 2.3 mM; Vmax, 2.3 x 10(6) units/mg). The in vitro stability of mutant enzymes at neutral pH was significantly reduced (S65T, 4% of normal; S65A, 29%; E66D, 54%). The intracellular alpha-Gal A activities of S65T, S65A, and E66D in COS1 cells transfected with corresponding plasmid DNAs were markedly lower than the normal enzyme activity (9, 26, and 68% of normal, respectively). However, intracellular enzyme activities were enhanced to 34% (S65T), 44% (S65A), and 80% (E66D) of normal, respectively, by cultivation of the cells with 20 microM 1-deoxygalactonojirimycin (a potent inhibitor of alpha-Gal A) for 24 h. These results suggest that Ser-65 is responsible for the stability of alpha-Gal A but not for the enzyme function. PMID- 10845699 TI - cDNA cloning of brevinase, a heterogeneous two-chain fibrinolytic enzyme from Agkistrodon blomhoffii brevicaudus snake venom, by serial hybridization polymerase chain reaction. AB - Brevinase is a heterogeneous two-chain fibrinolytic enzyme, different from all of the known single-chain enzymes. A cDNA encoding brevinase was cloned from the venom gland of Agkistrodon blomhoffii brevicaudus by serial hybridization-PCR. Serial hybridization-PCR effectively amplified the complete cDNA of brevinase from the mixture of closely related transcripts. The cDNA sequence of 744 nucleotides was determined. The cDNA sequence included an open reading frame of 233 amino acids composed of an A chain (77 residues) and a B chain (156 residues). The deduced amino acid sequence included a potential N-glycosylation site (N54-X-S56), O-glycosylation site (Ser179), and RGD sequence. Brevinase included a unique Arg77 residue at the C-terminus of the A chain, distinguishing it from all of the compared homologous single-chain proteases. It could be assumed that a posttranslational cleavage site is located between Arg77 and Asn78. Based on the sequence similarity to those of the venom proteases, we could deduce that the critical catalytic residues are His40, Asn78, Asp85, and Ser179 and that the six potential disulfide bonds are Cys7-Cys138, Cys26-Cys41, Cys73 Cys231, Cys117-Cys185, Cys149-Cys164, and Cys175-Cys200. Despite the conservation of critical sequences, the phylogenetic tree showed that two-chain brevinase might be evolved separately from the homologous single-chain proteases. PMID- 10845700 TI - Cleavage of vimentin by different retroviral proteases. AB - Proteases (PRs) of retroviruses cleave viral polyproteins into their mature structural proteins and replication enzymes. Besides this essential role in the replication cycle of retroviruses, PRs also cleave a variety of host cell proteins. We have analyzed the in vitro cleavage of mouse vimentin by proteases of human immunodeficiency virus type 1 (HIV-1) and type 2 (HIV-2), bovine leukemia virus (BLV), Mason-Pfizer monkey virus (M-PMV), myeloblastosis associated virus (MAV), and two active-site mutants of MAV PR. Retroviral proteases display significant differences in specificity requirements. Here, we show a comparison of substrate specificities of several retroviral proteases on vimentin as a substrate. Vimentin was cleaved by all the proteases at different sites and with different rates. The results show that the physiologically important cellular protein vimentin can be degraded by different retroviral proteases. PMID- 10845701 TI - Changes in glycosylation of human bile-salt-stimulated lipase during lactation. AB - Bile-salt-stimulated lipase (BSSL) is an enzyme in human milk, which is important for the fat digestion in the newborn infant. BSSL is highly glycosylated and includes one site for N-glycosylation and several sites for O-glycosylation. BSSL has previously been found to express Lewis a, Lewis b, and Lewis x carbohydrate antigens. In this study, glycosylation of BSSL was studied at different times during lactation. BSSL was purified from milk collected individually from four donors at several different times during the first 6 months of lactation. The BSSL glycans were characterized through monosaccharide analysis, high-pH anion exchange chromatography, matrix-assisted laser desorption-ionization mass spectrometry, and ELISA. Both total carbohydrate content and relative amount of sialic acid were higher in BSSL from the first lactation month as compared to BSSL from milk collected later in lactation. BSSL from the first lactation month also showed a different composition of sialylated O-linked glycans and the N linked oligosaccharides consisted of lower amounts of fucosylated structures compared to later in lactation. We also found a gradual increase in the expression of the carbohydrate epitope Lewis x on BSSL throughout the lactation period. This study shows that glycosylation of BSSL is dependent on blood group phenotype of the donor and changes substantially during the lactation period. PMID- 10845702 TI - Distribution and inducibility by 3-methylcholanthrene of family 1 UDP glucuronosyltransferases in the rat gastrointestinal tract. AB - UDP-Glucuronosyltransferases (UGT) catalyze the glucuronidation of a broad spectrum of endobiotic and xenobiotic substrates. The resulting glucuronides are more hydrophilic, facilitating renal and biliary excretion. Apart from hepatic glucuronidation, high rates of gastrointestinal glucuronidation have been observed. The aim of this study was to characterize the expression of family 1 UGTs (UGT1A) in liver, kidney, and all parts of the rat gastrointestinal tract by reverse transcription polymerase reaction (RT-PCR), Northern blot, and xenobiotic induction experiments. RT-PCR experiments were performed with primers specific for all known rat UGT1A mRNAs. UGT1A1, UGT1A6, and UGT1A7 were expressed in liver, kidney, and the gastrointestinal tract. UGT1A5 transcripts were detected in liver, but not in kidney or gastrointestinal tissue. In contrast, UGT1A2 and UGT1A3 were not expressed in liver or kidney, but were detected in intestine. Low levels of UGT1A3 were detectable in duodenum and jejunum. UGT1A2 was abundantly expressed in the small intestine; expression levels in the stomach and the large intestine were low. Quantitative evaluation of RNA levels by Northern blot revealed expression in gradients, with highest UGT1A mRNA levels in duodenum and decreasing levels in the small and large intestine. Only UGT1A6 was expressed at high levels in the rectum. Rats treated with 3-methylcholanthrene (3-MC) displayed a 10-fold induction of hepatic UGT1A6 and UGT1A7 mRNAs. In gastric tissues and in intestine, induction was 4-fold and 2-fold, respectively. In contrast to the constitutive expression of UGT1A7 in kidney, UGT1A6 was inducible in the liver. Effects of 3-MC on UGT1A1 expression revealed downregulation in the liver and highly variable effects in duodenum and stomach. This study demonstrates tissue-specific expression and tissue-specific induction patterns in rat liver, kidney, and gastrointestinal tract, which may represent the physiological basis of tissue-specific glucuronidation in rats. PMID- 10845703 TI - The quantitative oxidation of methionine to methionine sulfoxide by peroxynitrite. AB - Both peroxynitrous acid and peroxynitrite react with methionine, k(acid) = (1.7 +/- 0.1) x 10(3) M(-1) s(-1) and k(anion) = 8.6 +/- 0.2 M(-1) s(-1), respectively, and with N-acetylmethionine k(acid) = (2.8 +/- 0.1) x 10(3) M(-1) s(-1) and k(anion) = 10.0 +/- 0.1 M(-1) s(-1), respectively, to form sulfoxides. In contrast to the results of Pryor et al. (1994, Proc. Natl. Acad. Sci. USA 91, 11173-11177), a linear correlation between k(obs) and [met] was obtained. Surprisingly, for every two sulfoxides and nitrites formed, one peroxynitrite is converted to nitrate. Thus, methionine also catalyzes the isomerization of peroxynitrite to nitrate. Neither the pH nor the concentration of methionine affected the distribution of the yields of nitrite, nitrate, and methionine sulfoxide, which were the only products detected. No products other than nitrite, nitrate, and methioninesulfoxide could be detected. The reactions of methionine and N-acetylmethionine with peroxynitrous acid and peroxynitrite are simple bimolecular reactions that do not involve an activated form of peroxynitrous acid or of peroxynitrite. Nitrite, produced together with methionine sulfoxide, or present as a contamination in the peroxynitrite preparation, is not innocuous, but oxidizes methionine by one electron, which leads to the formation of methional and ethylene. PMID- 10845705 TI - A free cysteine residue at the dimer interface decreases conformational stability of Xenopus laevis copper,zinc superoxide dismutase. AB - The two Cu,Zn superoxide dismutases from the amphibian Xenopus laevis (denoted XSODA and XSODB) display different heat sensitivities, XSODA being more thermolabile than XSODB. In this study, we have investigated the contribution of a free cysteine residue located close to the subunit interface of XSODA to its lower thermal stability. We have found that mutation of residue Cys 150 to Ala in XSODA makes the thermal stability of this enzyme comparable to that of the wild type XSODB isoenzyme, while the introduction of a cysteine residue in the same position of XSODB renders this enzyme variant much more heat-sensitive. Differential scanning calorimetry experiments showed that XSODA has a melting temperature about 8.5 degrees C lower than that of XSODB. On the contrary, the melting temperature of XSODACys150Ala is very close to that of XSODB, while the melting temperature of XSODBSer150Cys is even lower than that of wild-type XSODA. These data indicate that the free cysteine residue present in XSODA affects not only the reversibility of unfolding of the enzyme but also its conformational stability. We suggest that the large effect of the Cys 150 residue on XSODA stability might be due to incorrect disulfide bond formation or disulfide bond interchange during heat-induced unfolding rather than to alteration of the interaction between the enzyme subunits. PMID- 10845704 TI - The mouse vitamin D receptor is mainly expressed through an Sp1-driven promoter in vivo. AB - The availability of the mouse vitamin D receptor (mVDR) gene has allowed a characterization of a TATA-less promoter containing a cluster of four Sp1 sites named Sp1-1, Sp1-2, Sp1-3, and Sp1-4 (F. Jehan and H. F. DeLuca, 1997, Proc. Natl. Acad. Sci. USA 94, 10138-10143). By means of primer extension analysis, S1 nuclease mapping and ribonuclease protection assay, the start site has been deduced, as has the existence of other minor transcription start sites. Initiation of transcription at the major site is located 4 bp upstream of the 5' end of the mVDR cDNA sequence and very close to the putative Sp1 sites. A second minor promoter might exist between exon 1 and exon 2 of the mVDR gene. The nucleotide sequence of the Sp1 region is well conserved between the mouse, the human, and the chicken VDR genes, suggesting an important role for these Sp1 sites. Gel shift analysis of the four Sp1 sites of the mVDR promoter has confirmed specific binding complexes to Sp1-1, Sp1-2, and Sp1-4 (Sp1-3 rather binds an unknown complex that is unable to bind the canonical Sp1 GGGGCGGGGC). Deletion or mutation of all the Sp1 sites eliminates promoter activity. However, mutation or deletion of individual Sp1 sites did not dramatically change the promoter activity, except for mutation of Sp1-3 that increases promoter activity. We, therefore, conclude that the mVDR promoter is controlled by the Sp1 sites and is the main VDR promoter in intestine and kidney. PMID- 10845706 TI - A snake venom inhibitor to muscarinic acetylcholine receptor (mAChR): isolation and interaction with cloned human mAChR. AB - An inhibitor to the muscarinic acetylcholine receptor (mAChR) was purified from the venom of Crotalus atrox (western diamondback rattlesnake). The inhibitor was found to be a 30-kDa homodimer protein with phospholipase A2 activity. In order to determine the subtype selectivity of the purified inhibitor, the inhibitory effect on the binding of two orthosteric antagonists, [3H]quinuclidinyl benzilate ([3H]QNB) and [3H]N-methylscopolamine methyl chloride ([3H]NMS), to five subtypes of cloned human mAChR was tested. The purified inhibitor reduced the binding of [3H]QNB and/or [3H]NMS to all subtypes of the mAChR while showing the highest inhibitory effect on the M5 subtype. The Kd values of the receptors for the antagonists were increased in the presence of the inhibitor; however, the Bmax values were not changed. The effects of the purified inhibitor on the dissociation of [3H]NMS from the receptors were also investigated. Dissociation of the antagonist was remarkably slowed down by addition of the inhibitor. These findings may suggest an allosteric action of the purified inhibitor. In addition, the present study indicates that the presence of mAChR inhibitors is quite common in snake venoms. PMID- 10845707 TI - Effects of nitric oxide on the copper-responsive transcription factor Ace1 in Saccharomyces cerevisiae: cytotoxic and cytoprotective actions of nitric oxide. AB - Previous studies indicate that nitric oxide (NO) can serve as a regulator/disrupter of metal-metabolizing systems in cells and, indeed, this function may represent an important physiological and/or pathophysiological role for NO. In order to address possible mechanisms of this aspect of NO biology, the effect of NO on copper metabolism and toxicity in the yeast Saccharomyces cerevisiae was examined. Exposure of S. cerevisiae to NO resulted in an alteration of the activity of the copper-responsive transcription factor Acel. Low concentrations of the NO donor DEA/NO were found to slightly enhance copper mediated activation of Acel. Since Acel regulates the expression of genes responsible for the protection of S. cerevisiae from metal toxicity, the effect of NO on the toxicity of copper toward S. cerevisiae was also examined. Interestingly, low concentrations of NO were also found to protect S. cerevisiae against the toxicity of copper. The effect of NO at high concentrations was, however, opposite. High concentrations of DEA/NO inhibited copper-mediated Acel activity. Correspondingly, high concentrations of DEA/NO (1 mM) dramatically enhanced copper toxicity. An intermediate concentration of DEA/NO (0.5 mM) exhibited a dual effect, enhancing toxicity at lower copper concentrations (<0.5 mM) and protecting at higher (> or =0.5 mM) copper concentrations. Thus, it is proposed that the ability of NO to both protect against (at low concentrations) and enhance (at high concentration) copper toxicity in S. cerevisiae is, at least partially, a result of its effect on Acel. The results of this study have implications for the role of NO as a mediator of metal metabolism. PMID- 10845708 TI - Ripple phases induced by alpha-tocopherol in saturated diacylphosphatidylcholines. AB - The effect of alpha-tocopherol on the structure and phase behavior of dilauroyl-, dimyristoyl-, dipalmitoyl-, and distearoyl-phosphatidylcholines was examined using X-ray diffraction and freeze-fracture electron microscopic methods. A ripple phase was observed in all of the mixtures at temperatures well below the pretransition temperature of the corresponding pure phospholipid. Freeze-fracture studies indicated that with proportion of alpha-tocopherol less than 5 mol% a ripple phase with large periodicity (50-150 nm) predominated and with about 10 mol% alpha-tocopherol a ripple phase of periodicity about 16 nm was formed. With more than 10 mol% alpha-tocopherol planar bilayers tended to be formed. Partial phase diagrams of mixed aqueous dispersions of saturated phosphatidylcholines and alpha-tocopherol over temperature ranges about the gel to liquid-crystal phase boundary have been constructed. Alpha-tocopherol-enriched domains form ripple phases that coexist with regions of lamellar gel phase of the pure phospholipid in mixtures containing less than 10 mol% alpha-tocopherol. The presence of increasing amounts of alpha-tocopherol in the phospholipid causes an increase in the proportion of ripple phase at the expense of pure phospholipid bilayer indicating that the alpha-tocopherol-enriched domains might possess a defined stoichiometry of the two constituents. PMID- 10845709 TI - A biophysical study of the interaction of the lipopeptide antibiotic iturin A with aqueous phospholipid bilayers. AB - Iturin A is a lipopeptide extracted from the culture media of Bacillus subtilis which shows a strong antifungal action. The interaction of iturin A with multilamellar vesicles of dimyristoylphosphatidylcholine (DMPC) induced structures which did not sediment during centrifugation. Electron microscopy after negative staining showed that, at 30 mol%, iturin A/DMPC vesicles were visible but smaller than those formed by pure DMPC. Thermograms of DMPC/iturinA obtained after differential scanning calorimetry, at low concentrations of iturin A, were interpreted as indicating the presence of two laterally separated phases, one formed by pure phospholipid and the other by lipopeptide-phospholipid complexes, these two separated phases being already detected even at low concentrations such as 2 mol%. Fluorescence quenching experiments showed that the D-Tyr residue of the lipopeptide was fully accessible to the aqueous medium, indicating that the polar part of iturin A is located outside of the membrane hydrophobic palisade. It was concluded that the membrane barrier properties are likely to be damaged in the area where the lipid complexes are accumulated, due to structural fluctuations, and this may be one of the bases of its biological activity. Iturin-A was also able to greatly destabilize dielaidoylphosphatidylethanolamine (DEPE) membranes in the fluid form, producing a new structure which had a poor correlation in X-ray diffraction, and in 31P NMR spectroscopy gave rise to a spectrum containing a double isotropic signal. Iturin A was shown to induce DEPE to adopt phases other than H(II) inverted hexagonal, underlining that this lipopeptide is capable of modifying the curvature of the membrane, which may also be important in explaining the tendency of iturin A to create small vesicles and which may be another of the bases of its biological activity. PMID- 10845710 TI - The amino-terminal region of the long-chain fatty acid transport protein FadL contains an externally exposed domain required for bacteriophage T2 binding. AB - The fatty acid transport protein FadL from Escherichia coli is predicted to be rich in beta-structure and span the outer membrane multiple times to form a long chain fatty acid specific channel. Proteolysis of FadL within whole cells, total membranes, and isolated outer membranes identified two trypsin-sensitive sites, both predicted to be in externally exposed loops of FadL. Amino acid sequence analysis of the proteolytic fragments determined that the first followed R93 and yielded a peptide beginning with 94S-L-K-A-D-N-I-A-P-T-A104 while the second followed R384 and yielded a peptide beginning with 385S-I-S-I-P-D-Q-D-R-F-W395. Proteolysis using trypsin eliminated the bacteriophage T2 binding activity associated with FadL, suggesting the T2 binding domain within FadL requires elements within one of these extracellular loops. A peptide corresponding to the amino-terminal region of FadL (FadL28-160) was purified and shown to inactivate bacteriophage T2 in a concentration-dependent manner, supporting the hypothesis that the amino-proximal extracellular loop of the protein confers T2 binding activity. Using an artificial neural network (NN) topology prediction method in combination with Gibbs motif sampling, a predicted topology of FadL within the outer membrane was developed. According to this model, FadL spans the outer membrane 20 times as antiparallel beta-strands. The 20 antiparallel beta-strands are presumed to form a beta-barrel specific for long-chain fatty acids. On the basis of our previous studies evaluating the function of FadL using site-specific mutagenesis of the fadL gene, proteolysis of FadL within outer membranes, and studies using the FadL28-160 peptide, the predicted extracellular regions between beta-strands 1 and 2 and beta-strands 3 and 4 are expected to contribute to a domain of the protein required for long-chain fatty acid and bacteriophage T2 binding. The first trypsin-sensitive site (R93) lies between predicted beta strands 3 and 4 while the second (R384) is between beta-strands 17 and 18. The trypsin-resistant region of FadL is predicted to contain 13 antiparallel beta strands and contribute to the long-chain fatty acid specific channel. PMID- 10845711 TI - Carbonic anhydrase III: the phosphatase activity is extrinsic. AB - The carbonic anhydrases reversibly hydrate carbon dioxide to yield bicarbonate and hydrogen ion. They have a variety of physiological functions, although the specific roles of each of the 10 known isozymes are unclear. Carbonic anhydrase isozyme III is particularly rich in skeletal muscle and adipocytes, and it is unique among the isozymes in also exhibiting phosphatase activity. Previously published studies provided evidence that the phosphatase activity was intrinsic to carbonic anhydrase III, that it had specificity for tyrosine phosphate, and that activity was regulated by reversible glutathionylation of cysteine186. To study the mechanism of this phosphatase, we cloned and expressed the rat liver carbonic anhydrase III. The purified recombinant had the same specific activity as the carbonic anhydrase purified from rat liver, but it had virtually no phosphatase activity. We attempted to identify an activator of the phosphatase in rat liver and found a protein of approximately 14 kDa, the amount of which correlated with the phosphatase activity of the carbonic anhydrase III fractions. It was identified as liver fatty acid binding protein, which was then purified to test for activity as an activator of the phosphatase and for protein-protein interaction, but neither binding nor activation could be demonstrated. Immunoprecipitation experiments established that carbonic anhydrase III could be separated from the phosphatase activity. Finally, adding additional purification steps completely separated the phosphatase activity from the carbonic anhydrase activity. We conclude that the phosphatase activity previously considered to be intrinsic to carbonic anhydrase III is actually extrinsic. Thus, this isozyme exhibits only the carbon dioxide hydratase and esterase activities characteristic of the other mammalian isozymes, and the phosphatase previously shown to be activated by glutathionylation is not carbonic anhydrase III. PMID- 10845712 TI - Solubilization, partial purification, and characterization of a fatty aldehyde decarbonylase from a higher plant, Pisum sativum. AB - Enzymatic decarbonylation of fatty aldehydes generates hydrocarbons. The particulate enzyme that catalyzes the decarbonylation has not been solubilized and purified from any organism but a green alga. Here we report the solubilization, purification, and partial characterization of the decarbonylase from a higher plant. Decarbonylase from a particulate preparation from pea (Pisum sativum) leaves, enriched in decarbonylase, was solubilized with beta-octyl glucoside and partially purified. SDS-PAGE showed a major protein band at 67 kDa. Rabbit antibodies raised against this protein specifically cross-reacted with the 67-kDa protein in solubilized microsomal preparations; anti-ribulose bisphosphate carboxylase cross-reacted only with the 49-kDa large subunit of the carboxylase, but not with any protein near 67 kDa, showing the absence of any contamination from cross-linked small-large subunit of the carboxylase found in the green algal enzyme preparation. Anti-67-kDa protein antibodies inhibited decarbonylation catalyzed by the enzyme preparations, showing that this protein represents the decarbonylase. Decarbonylase activity of the purified enzyme required phospholipids for activity; phosphatidylcholine was the preferred lipid although phosphatidylserine and phosphatidylethanolamine could substitute less effectively. Half-maximal activity was observed at 40 microM octadecanal. The purified enzyme produced alkane and CO and was inhibited by O2, NADPH, and DTE. Metal ion chelators severely inhibited the enzyme and Cu2+ fully restored the enzyme activity. Purified enzyme preparations consistently showed the presence of Cu, and copper protoporphyrin IX catalyzed decarbonylation. These results suggest that this higher plant enzyme probably is a Cu enzyme unlike the green algal enzyme that was found to have Co. PMID- 10845713 TI - Tyrosine nitration of c-SRC tyrosine kinase in human pancreatic ductal adenocarcinoma. AB - During pancreatic tumorigenesis, the equilibrium between cell survival and cell death is altered, allowing aggressive neoplasia and resistance to radiation and chemotherapy. Local oxidative stress is one mechanism regulating programmed cell death and growth and may contribute to both tumor progression and suppression. Our recent in situ immunohistochemical studies demonstrated that levels of total nitrotyrosine, a footprint of the reactive nitrogen species peroxynitrite, are elevated in human pancreatic ductal adenocarcinomas. In this study, quantitative HPLC-EC techniques demonstrated a 21- to 97-fold increase in the overall levels of nitrotyrosine of human pancreatic tumor extracts compared to normal pancreatic extracts. Western blot analysis of human pancreatic tumor extracts showed that tyrosine nitration was restricted to a few specific proteins. Immunoprecipitation coupled with Western analysis identified c-Src tyrosine kinase as a target of both tyrosine nitration and tyrosine phosphorylation. Peroxynitrite treatment of human pancreatic carcinoma cells in vitro resulted in increased tyrosine nitration and tyrosine phosphorylation of c-Src kinase, increased (>2-fold) c-Src kinase activity, and increased association between c-Src kinase and its downstream substrate cortactin. Collectively, these observations suggest that peroxynitrite-mediated tyrosine nitration and tyrosine phosphorylation of c-Src kinase may lead to enhanced tyrosine kinase signaling observed during pancreatic ductal adenocarcinoma growth and metastasis. PMID- 10845714 TI - III. Instantaneous inhibition by compound 48/80 of tissue factor-initiated extrinsic coagulation is mediated by the downregulation of factor VII activation. AB - Our previous study has demonstrated a unique biological function of compound 48/80 (48/80) in the downregulation of monocytic tissue factor (TF)-initiated hypercoagulation in response to bacterial endotoxin (lipopolysaccharide, LPS) [A. J. Chu et al. (1999) Biochim. Biophys. Acta 1472, 386-395]. The inhibition was not due to the blockade of LPS cell signaling as evidenced by the unaffected LPS induced TF synthesis. In the present study, we investigate the direct inhibitory action of 48/80 on the extrinsic coagulation cascade. TF-initiated coagulation was assayed by a single-stage clotting assay. Chromogenic assays dissected the extrinsic pathway to measure the activities of FVII, FX, and prothrombin by monitoring the hydrolyses of nitroaniline-conjugated substrates, identifying the inhibitory site(s). We report that 48/80 in vitro instantaneously inhibited rabbit brain thromboplastin (rbTF)-initiated coagulation in a dose-dependent manner. 48/80 preferentially inhibited FVII activation without any detectable effect on FVIIa, FXa, and thrombin activities. Neither FX activation nor prothrombin activation was affected. The significant inhibition on FVII activation was found to be noncompetitive with a fourfold reduction in the apparent Vmax of FVIIa formation from 7.1 to 1.7 nM/min, while the apparent Km (approximately 365 nM) remained unaffected. Western blotting analysis further confirmed that FVIIa formation derived from FVII was significantly diminished by 48/80, which was accompanied by blocked FVII binding to rbTF. In conclusion, 48/80 readily blocked FVII binding to rbTF, leading to diminished FVII activation and FVIIa formation. As a result, TF-initiated extrinsic coagulation was downregulated. PMID- 10845715 TI - Limited proteolysis of branching enzyme from Escherichia coli. AB - Branching enzyme is involved in determining the structure of starch and glycogen. It catalyzes the formation of branch points by cleavage and transfer of alpha-1,4 glucan chains to alpha-1,6 branch points. Branching enzyme belongs to the amylolytic family of enzymes containing four conserved regions in a central (alpha/beta)8-barrel. Limited proteolysis of the branching enzyme from Escherichia coli (84 kDa) by proteinase K produced a truncated protein of 70-kDa, which still retained 40-60% of branching activity, depending on the type of assay used. Amino acid sequencing showed that the 70-kDa protein lacked 111 or 113 residues at the amino terminal, whereas the carboxy terminal was still intact. We purified this truncated enzyme to homogeneity and analyzed its properties. The enzyme had a three- to fourfold lower catalytic efficiency than the native enzyme, whereas the substrate specificity was unaltered. Furthermore, a branching enzyme with 112 residues deleted at the amino terminal was constructed by recombinant technology and found to have properties identical to those of the proteolyzed enzyme. PMID- 10845716 TI - Identification, characterization, and immunolocalization of a nucleoside triphosphate diphosphohydrolase in pig liver. AB - Different isoforms of nucleoside triphosphate diphosphohydrolases (NTPDases; EC 3.6.1.5), also identified as ATP diphosphohydrolases, have been previously described in mammalian tissues. We report here the biochemical characterization of NTPDases in the pig liver. Optimum pH of catalysis is more acidic for this enzyme than for NTPDases (neutral or alkaline pH) found in other mammalian tissues. It is less sensitive to bile salts than the bovine spleen NTPDase. Calculated Km values for ATP and ADP (31 and 21 microM, respectively) are slightly higher than those reported for the latter enzyme. Electrophoretograms of these enzymes also show different migration patterns. Western blots with Ringo, an antibody that recognizes the different isoforms of mammalian NTPDases, show a small but reproducible difference in estimated molecular masses (75 kDa for liver vs 78 kDa for spleen NTPDase). A second antibody, generated against a different sequence of NTPDase I, does not recognize the liver enzyme, thereby indicating some differences in primary structure. Immunolocalization produced a strong signal on hepatocytes, epithelial cells of the bile duct system, and vascular cells. Immunoreactivity was variable among hepatocytes of different lobules and among hepatocytes within a given lobule. In general, those located in the perilobular zone were more reactive than those located in the central zone and in the periphery of the centrolobular vein. PMID- 10845717 TI - A tale of two trials: selectively replicating herpesviruses for brain tumors. PMID- 10845718 TI - Anti-inflammatory action of type I interferons deduced from mice expressing interferon beta. AB - Type I interferons (IFN) are widely used for the therapeutic treatment of viral infections, tumor growth and various chronic diseases such as multiple sclerosis. Antagonism between type I IFNs and IFN-gamma has been described in cells of the immune system, in particular in the activation of macrophages. To study the systemic effects of type I IFNs we used transgenic mice carrying a human IFN-beta (hIFN-beta) gene under the control of the rat insulin I promoter. These animals expressed high levels of hIFN-beta in beta-pancreatic cells, and the ability of the macrophages to respond to pro-inflammatory stimuli was analyzed. Transgenic mice exhibited an increased extravasation of cells to the peritoneal cavity after eliciting with thioglycollate broth. The expression of the inducible form of nitric oxide synthase and cyclooxygenase-2, two enzymes involved in inflammation, was impaired in transgenic animals challenged with lipopolysaccharide and IFN gamma. Analysis of the mechanisms leading to this attenuated inflammatory response showed a decrease in the serum levels of TNF-alpha and an inhibition of the activation of the transcription factor NF-KB in various tissues. These results indicate that systemic administration of IFN-beta might influence the response to pro-inflammatory stimuli, in particular through the antagonism of IFN gamma signaling. PMID- 10845719 TI - Electrically mediated plasmid DNA delivery to hepatocellular carcinomas in vivo. AB - Gene therapy by direct delivery of plasmid DNA has several advantages over viral gene transfer, but plasmid delivery is less efficient. In vivo electroporation has been used to enhance delivery of chemotherapeutic agents to tumors in both animal and human studies. Recently, this delivery technique has been extended to large molecules such as plasmid DNA. Here, the successful delivery of plasmids encoding reporter genes to rat hepatocellular carcinomas by in vivo electroporation is demonstrated. PMID- 10845720 TI - Efficiency of transgenic T cell generation from gene-marked cultured human CD34+ cord blood cells is determined by their maturity and the cytokines present in the culture medium. AB - Success of gene therapy for diseases affecting the T cell lineage depends on the thymic repopulation by genetically engineered hematopoietic progenitor cells (HPC). Although it has been shown that retrovirally transduced HPC can repopulate the thymus, little information is available on the effect of the culture protocol. Moreover, for expansion of the number of HPC, cytokine supplemented culture is needed. Here, we transduced purified human umbilical cord blood (CB) CD34+ cells in cultures supplemented with various combinations of the cytokines thrombopoietin (TPO), stem cell factor (SCF), flt3/flk-2 ligand (FL), interleukin 3 (IL-3) and IL-6, and investigated thymus-repopulating ability of gene-marked HPC in vitro. Irrespective of the cytokine cocktail used, transduced CD34+CD38- CB cells, expressing the marker green fluorescent protein (GFP) encoded by the MFG-GFP retrovirus, have both superior proliferative and thymus-repopulating potential compared with transduced CD34+CD38+ CB cells. Effectively transduced GFP+CD34+CD38- HPC, cultured for 3 or 17 days, more readily generated T cells than GFP- HPC from the same culture. The reverse was true in the case of CD34+CD38+ HPC cultures. Finally, our results indicate that the number of GFP+ T cell progenitors actually increased during culture of CD34+CD38- HPC, in a magnitude that is determined by the cytokine cocktail used during culture. PMID- 10845721 TI - Tumor cells expressing membrane-bound form of IL-4 induce antitumor immunity. AB - Local cytokine concentrations are required for inhibition of tumor growth with less toxic side-effects. However, genetically engineered tumor cells secreting cytokines still induce toxicity and activate bystander cells. To circumvent such problems, membrane-bound forms of IL-4 (IL-4m) were expressed on MethA fibrosarcoma tumor cells. Chimeric forms of IL-4 with the type I transmembrane protein CD4 or type II transmembrane protein TNF were designed to express IL-4 in opposite orientations on the tumor cell surface. The IL-4m on tumor clones was able to support cell growth of the IL-4 dependent cytotoxic cell line (CT.4S) and the Th2 cell clone (D10). Furthermore, the IL-4m tumor clones stimulated proliferation of 2C TCR transgenic spleen cells which are responsive to Ld MHC class I molecules. Expression of the IL-4/TNF chimeric protein on MethA cells elicited antitumor immunity and protected from MethA tumor challenge. The proposed tumor vaccine may serve as an effective gene therapy method to avoid the toxicity of recombinant cytokines and bulk bystander leukocyte stimulation encountered in conventional cytokine gene therapy. PMID- 10845722 TI - Intravesical liposome-mediated interleukin-2 gene therapy in orthotopic murine bladder cancer model. AB - Using a novel orthotopic MBT-2 murine bladder tumor model, we evaluated the feasibility of intravesical gene therapy utilizing a cationic liposome, DMRIE/DOPE. Superficial bladder tumors were consistently established by intravesical instillation of 5x10(5) MBT-2 cells in syngeneic C3H female mice. In situ gene transfer to bladder tumors was accomplished via intravesical instillation of plasmid DNA/DMRIE/DOPE lipoplex. Beta-Galactosidase (beta-gal) gene expression was preferentially evident in bladder tumors and was present for at least 7 days after a single 30 min in situ transfection. Murine interleukin-2 (IL-2) gene was used for treatment of 3-day-old pre-established bladder tumors. Forty percent of animals treated with IL-2 gene were completely free of tumors by 60 days following the initial tumor implantation, while all control groups treated with beta-gal gene died. Those animals initially cured of pre-established tumors were completely resistant to a subsequent tumor re-challenge and their splenocyte-derived cytotoxic T lymphocytes were shown to be specific to MBT-2 cells, indicating that immunological memory against MBT-2 tumors was elicited by the treatment. These results demonstrate the possibility of an effective clinical application of this in situ intravesical IL-2 gene delivery system to high-risk superficial bladder tumors, obviating a need for tumor procurement and ex vivo gene transfer. PMID- 10845723 TI - Antisense ATM gene therapy: a strategy to increase the radiosensitivity of human tumors. AB - Atm, the gene mutated in ataxia-telangiectasia (AT) patients, is an essential component of the signal transduction pathway that responds to DNA damage due to ionizing radiation (IR). We attenuated ATM protein expression in human glioblastoma cells by expressing antisense RNA to a functional domain of the atm gene. While ATM expression decreased, constitutive expression of p53 and p21 increased. Irradiated ATM-attenuated cells failed to induce p53, demonstrated radioresistant DNA synthesis, and increased radiosensitivity. Antisense-ATM gene therapy in conjunction with radiation therapy may provide a novel strategy for the treatment of cancer. PMID- 10845724 TI - Toxicity evaluation of replication-competent herpes simplex virus (ICP 34.5 null mutant 1716) in patients with recurrent malignant glioma. AB - The herpes simplex virus (HSV) ICP34.5 null mutant 1716 replicates selectively in actively dividing cells and has been proposed as a potential treatment for cancer, particularly brain tumours. We present a clinical study to evaluate the safety of 1716 in patients with relapsed malignant glioma. Following intratumoural inoculation of doses up to 10(5) p.f.u., there was no induction of encephalitis, no adverse clinical symptoms, and no reactivation of latent HSV. Of nine patients treated, four are currently alive and well 14-24 months after 1716 administration. This study demonstrates the feasibility of using replication competent HSV in human therapy. PMID- 10845725 TI - Conditionally replicating herpes simplex virus mutant, G207 for the treatment of malignant glioma: results of a phase I trial. AB - G207 is a conditionally replicating derivative of herpes simplex virus (HSV) type 1 strain F engineered with deletions of both gamma(1)34.5 loci and a lacZ insertion disabling the UL39 gene. We have demonstrated the efficacy of G207 in treating malignant glial tumors in athymic mice, as well as the safety of intracerebral G207 inoculation in mice and in Aotus nancymai. We sought to determine the safety of G207 inoculation into cerebral malignant glial tumors in humans. Criteria for inclusion into this dose-escalation study were the diagnosis of histologically proven malignant glioma, Karnofsky score > or = 70, recurrence despite surgery and radiation therapy, and an enhancing lesion greater than 1 cm in diameter. Serial magnetic resonance images were obtained for volumetric analysis. The trial commenced at a dose of 10(6) plaque forming units (p.f.u.) inoculated at a single enhancing site and was completed when the 21st patient was inoculated with 3x10(9) p.f.u. at five sites. While adverse events were noted in some patients, no toxicity or serious adverse events could unequivocally be ascribed to G207. No patient developed HSV encephalitis. We found radiographic and neuropathologic evidence suggestive of anti-tumor activity and long-term presence of viral DNA in some cases. PMID- 10845726 TI - Adenoviral vector which delivers FasL-GFP fusion protein regulated by the tet inducible expression system. AB - Fas ligand (FasL) is a member of the tumor necrosis family and when bound to its receptor, Fas, induces apoptosis. It plays important roles in immune response, degenerative and lymphoproliferative diseases, development and tumorigenesis. It is also involved in generation of immune privilege sites in the eye and testis. Harnessing the power of this molecule is expected to lead to a powerful chemotherapeutic. We describe the construction and characterization of replication-deficient adenoviral vectors that express a fusion of murine FasL and green fluorescent protein (GFP). FasL-GFP retains full activity of wild-type FasL, at the same time allowing for easy visualization and quantification in both living and fixed cells. The fusion protein is under the control of a tetracycline regulated gene expression system. Tight control of expression is achieved by creating a novel 'double recombinant' Ad vector, in which the tet-responsive element and the transactivator element are built into the opposite ends of the same vector to avoid enhancer interference. Expression can be conveniently regulated by tetracycline or its derivatives in a dose-dependent manner. The vector was able to deliver FasL-GFP gene to cells in vitro efficiently, and the expression level and function of the fusion protein was modulated by the concentration of doxycycline. This regulation allows us to produce high titers of the vector by inhibiting FasL expression in an apoptosis-resistant cell line. Induction of apoptosis was demonstrated in all cell lines tested. These results indicate that our vector is a potentially valuable tool for FasL-based gene therapy of cancer and for the study of FasL/Fas-mediated apoptosis and immune privilege. PMID- 10845727 TI - Efficient gene transfer to hematopoietic progenitor cells using SV40-derived vectors. AB - We used recombinant SV40 (rSV40)-derived vectors to deliver transgenes to human and simian hematopoietic progenitor cells in culture, and in vivo after transduction ex vivo. rSV40 are highly efficient vectors that are made in very high titers. They infect almost all cells, whether resting or dividing. Two rSV40s were used: SV(HBS), carrying hepatitis B surface antigen as a marker; and SV(Aw) carrying IN#33, a single chain Fv antibody against HIV-1 integrase. CD34+ cells derived from human fetal bone marrow (HFBM) and rhesus macaque bone marrow were transduced once with SV(HBS) without selection. On average 60% of colonies derived from transduced CD34+ cells carried and expressed HBsAg, as assessed by PCR and immunochemistry. Transgene carriage persisted following differentiation of transduced rhesus CD34+ cells into T lymphocytes. In an effort to increase the percentage of gene-marked cells, three sequential treatments of CD34+ cells were done using SV(Aw), without selection. Two weeks later, >95% of colonies expressed IN#33. Unselected SV(Aw)-transduced CD34+ cells from HFBM were transplanted into sublethally irradiated SCID mice. Bone marrow harvested 3 months later showed that >50% of bone marrow cells expressed IN#33. This is comparable with the percentage of human cells in these animals' bone marrow as judged by immunostaining for human CD45. The stability and longevity of transduction in this setting suggests that rSV40 vectors integrate into the cellular genome. This possibility was supported by finding that PCR of genomic DNA using primer pairs with one cellular and one viral primer yielded PCR products only in transduced, but not control, cells. These PCR products hybridized with an SV40 DNA fragment. Thus, rSV40 vectors transduce normal human and primate bone marrow progenitor cells effectively without selection, and maintain transgene expression in vivo following reimplantation. Such high efficiency transduction may be useful in treating diseases of CD34+ cells and their derivatives. PMID- 10845728 TI - Increasing endothelial cell specific expression by the use of heterologous hypoxic and cytokine-inducible enhancers. AB - One of the current challenges in gene therapy is to construct a vector that will target specific tissues. Targeting expression to endothelium is of particular interest in the treatment of several pathologies. We have shown previously that defined regions of the E-selectin and KDR promoters confer endothelial cell specific expression following retroviral delivery. However, the levels of expression were low. In an attempt to increase expression but to preserve the tissue specificity we have examined hypoxic and cytokine-inducible enhancer elements in combination with the KDR and E-selectin promoters. Both enhancers should be active in the tumour environment, boosting expression and giving additional specificity of gene expression in the tumour endothelium. The hypoxia response element (HRE) of the murine phosphoglycerate kinase-1 (PGK-1) promoter was used as a hypoxic enhancer and the tandem-binding site for NFKB from the murine vascular cell adhesion molecule-1 (VCAM-1) promoter as a cytokine inducible enhancer. The HRE conferred hypoxia inducibility to the KDR and E selectin promoters. Endothelial specificity of expression was retained with the KDR but not the E-selectin promoter. The NFKB-binding site conferred responsiveness to TNF-alpha to the KDR promoter, however the level of induction was less than that achieved with the HRE. Retrovirus combining both enhancer elements transferred inducibility by hypoxia and TNF-alpha, and reached the highest expression levels upon stimulation. These results confirm that heterologous enhancer elements may operate on a single endothelial cell specific promoter. These findings make the use of inducible enhancers a promising strategy for increasing tissue specific gene expression. PMID- 10845729 TI - Oxcarbazepine: a review. PMID- 10845730 TI - A multicentre, double-blind, randomized, parallel group study to evaluate the tolerability and efficacy of two oral doses of levetiracetam, 2000 mg daily and 4000 mg daily, without titration in patients with refractory epilepsy. AB - The aim of this study was to determine the tolerability and efficacy of two oral regimens of levetiracetam, 1000 mg and 2000 mg twice daily, as add-on treatment without titration in patients with refractory epilepsy. After a 1- to 4-week baseline, 119 patients were randomized to receive levetiracetam 2000 mg daily, 4000 mg daily, or placebo for a 24-week double-blind period, then levetiracetam 4000 mg daily in a 24-week open-label phase. Somnolence was the most common reason for discontinuation, and along with asthenia, occurred more frequently with levetiracetam than placebo. Responder rates were higher with levetiracetam 2000 mg and 4000 mg daily (48.1% [P < 0.05] and 28.6% [NS], respectively) than placebo (16.1%). In the open-label phase, the overall responder rate was 43.0%. Switching from placebo to levetiracetam increased the overall responder rate from 16.7% to 44.0%. No such increase was observed with patients initiated on levetiracetam 2000 mg daily. Levetiracetam initiated at doses of 2000 mg or 4000 mg daily without titration is well-tolerated and effective as add-on therapy in patients with partial and/or generalized seizures. The higher dose may be related to an increased incidence of somnolence and is not necessarily more effective than the lower dose. PMID- 10845731 TI - The contributing factors to medical cost of epilepsy: an estimation based on a French prospective cohort study of patients with newly diagnosed epileptic seizures (the CAROLE study). Active Coordination of the Longitudinal Observational Network in Epilepsy. AB - Epilepsy is a common neurological condition with significant resource implications. An estimation was performed in France of the direct medical cost of patients presented with newly diagnosed seizures and followed during the first two years after diagnosis. This estimation was based on the service utilization data collected from French prospective cohort study (CAROLE: 1942 patients enrolled). Costs were estimated in a societal perspective in 1998 value. The impact on the costs of different factors like age, aetiologic categorization and severity of seizures (type and number of seizures), and treatment by anti epileptic drugs (AEDs) was analysed. The mean annual direct epilepsy-related costs per patient were estimated to be 14 305 F and 3766 F for the first and the second year of follow-up respectively, 68% and 40% of the costs were devoted to inpatient care. Costs during the first year were highly sensitive to aetiologic categorization of seizures at inclusion and to other clinical parameters. Second year costs had a much lower variance and were sensitive to frequency of seizures and the fact of being treated or not by AEDs. Our data emphasize the importance of seizure control as means of reducing the costs of epilepsy especially during the first year of follow-up. PMID- 10845732 TI - Review on phytotherapy in epilepsy. PMID- 10845733 TI - Photosensitivity in juvenile myoclonic epilepsy. AB - Photosensitivity is reported to occur in approximately 40% of patients with juvenile myoclonic epilepsy. Our experience suggests that the prevalence is higher and may be related to both the duration of intermittent photic stimulation and also the age at which the procedure is undertaken. A two-year retrospective review of all EEGs was undertaken on all children attending a paediatric EEG department to identify those with juvenile myoclonic epilepsy. Photosensitivity was defined as a generalized spike or spike-wave paroxysm occurring at least twice during intermittent photic stimulation. Sixty-one children with a diagnosis of juvenile myoclonic epilepsy with a median age of 13 (range 7-16) years were identified, 55 (90%) of whom were photosensitive. Eighteen of these 55 patients showed photosensitivity only after four minutes of continuous photic stimulation. The prevalence of photosensitivity in juvenile myoclonic epilepsy is likely to be higher than previously reported. When a diagnosis of juvenile myoclonic epilepsy is being considered, the initial diagnostic EEG should include intermittent photic stimulation for up to five minutes, or less if the patient shows evidence of photosensitivity. The identification of photosensitivity may have important management implications. PMID- 10845734 TI - Long-term open multicentre, add-on trial of vigabatrin in adult resistant partial epilepsy. The Canadian Vigabatrin Study Group. AB - Vigabatrin (VGB) has been shown in a number of clinical trials with varying designs to be effective and well-tolerated as both add-on therapy and monotherapy in epilepsy with partial seizures with or without secondary generalization as well as in infantile spasms. The present study is an open, long-term (1 year) extension of a randomized double-blind placebo-controlled multicentre Canadian trial of VGB in resistant partial adult epilepsy. The present study was designed to examine the safety and long-term efficacy of VGB. Completers of the preceding double-blind study had their dose of VGB titrated to 4 g/day over 3 weeks. Patients were evaluated every 2-4 weeks and at week 14 were allowed to continue only if they achieved a 50% seizure reduction compared with pre-VGB baseline. In addition to neurological and physical examinations, safety was assessed by a cognitive psychosocial test battery, visual and somatosensory evoked potentials and MRI scans. Ninety-seven of 100 eligible patients entered the study, 53 of whom completed the 52 weeks. Fifty-eight percent of the patients had a greater than 50% seizure reduction in seizures vs. pre-VGB baseline. Seizure reductions of 56% and 45%, respectively, were seen in the VGB and placebo groups from the preceding study. Fifty-four percent of patients were judged by the investigators to have experienced at least a moderate therapeutic effect. Discontinuations were 29% for lack of efficacy and 12% for adverse effects. There was a mean weight gain of 3.7 +/- 0.2 kg by end of study. Neurologica/psychiatric side effects were the most common reason for withdrawal including three behavioral reactions attributed to the drug which required temporary hospitalization. There were no abnormalities on laboratory or special tests and there was a tendency for improvement on most tests of cognitive function and mood. Vigabatrin, as an add on agent, is well-tolerated and can be of long-term benefit in a substantial proportion of patients with intractable partial epilepsy. PMID- 10845735 TI - Case-control and qualitative study of attrition in a community epilepsy programme in rural India. AB - Dropout from epilepsy programmes is a serious problem in developing countries and has not been systematically studied before. We set up a community-based programme for children with epilepsy in rural India. The aim of this study was to assess reasons for dropout. We assessed medical and sociodemographic variables for their effect on dropout at 12 months using an unmatched case-control design on 32 cases and 62 controls. We also interviewed the parents of 32 children who dropped out of treatment, using a topic schedule. Two-thirds of the dropouts occurred within the first 6 months of treatment. Severely impaired children were more likely to drop out (odds ratio 4.60, 95% CI: 1.0-21.0) and families who had tried AEDs before were less likely to do so (odds ratio 0.12, 95% CI: 0.015-0.88). Denial of diagnosis, access problems and symptom resolution were the other main reasons underlying attrition. Active ascertainment methods should be reconsidered in community programmes. Very poor families without a male head or with long journey times are at high risk of dropout. People with severe impairments need appropriate integrated rehabilitation. PMID- 10845736 TI - The role of patient companions in long-term video-EEG monitoring. AB - In developing countries it is difficult to have full-time dedicated nurses in Epilepsy Monitoring Units (EMU). Our one-bed EMU is within the Neurology Service and is adequately staffed during daytime working hours only. So we created a new model where the patient's companion was asked to press a nurse call button, allowing the examination of the patient by the nurse. In this study we aimed to understand how patient companions behaved and which factors influenced their behaviour. Patients were allowed to choose a single companion who were educated by the specialist monitoring nurse according to a protocol. Only the first recorded seizures of the patients were included in the study. The seizures were reviewed from the video-cassette recordings and the behaviour of the companions was scored according to the results of the following three questions: (1) when was the seizure noticed?; (2) was the nurse call button pushed?; and (3) did the companion prevent the recording of the seizure by the camera? The companions were grouped according to the following criteria; age, sex, level of education, type of relationship. The scores were compared for each criterion separately. The behaviours of the 50 companions (34F, 16M; age: 25-72) were studied. When statistically compared for age, sex and level of education, there were no significant differences between different groups. However, the mean score of the 47 companions who were immediate family members (3.72) was greater than those three who were not (1.66) In one-bed EMUs, patient companions who are family members can help nurses in the early detection of seizures. PMID- 10845737 TI - Dietary practices and use of the ketogenic diet in the UK. AB - With the introduction of Internet communications, parental interest has increased in the use of the ketogenic diet for epilepsy. It was decided to audit current practice in the use of the ketogenic diet in the UK. All paediatric dietitians who were members of the Paediatric Group of the British Dietetic Association were surveyed by a postal questionnaire. There was a 51% response rate. Twenty-two hospitals (17%) used the ketogenic diet with 101 patients being treated. Fifty nine percent used the traditional 4:1 (four parts fat : one part carbohydrate, one part protein) classical ketogenic diet and 41% used the medium chain triglyceride diet (60% MCT fat). The age of patients ranged from 1 to over 11 years. There were wide variations in its application, with 66% of hospitals initiating the diet in hospital and 33% at home. The dietary energy administered varied from 60 to 90 kcal/kg/day, and there was no consistent policy on vitamin and mineral supplementation. Twenty-five patients continued to follow the diet after 12 months. Therefore, the ketogenic diet is commonly used for the treatment of intractable epilepsy in the UK. Further research work is needed on its nutritional safety and application. PMID- 10845738 TI - Efficacy of lamotrigine in institutionalized, developmentally disabled patients with epilepsy: a retrospective evaluation. AB - The paper evaluates the efficacy of the newer anticonvulsant lamotrigine in a developmentally disabled patient population. A retrospective evaluation was done at two institutional centres to assess adjunctive lamotrigine (Lamictal) efficacy in a developmentally disabled population. Mean seizure frequency was compared between a 2-month pre-lamotrigine baseline period and a 2-month treatment period. A 3-month lamotrigine titration phase occurred between baseline and treatment periods. Seizure frequency data was obtained from standardized, daily seizure records. Adverse effect data was obtained from medical and nursing notes. An intent to treat analysis was performed. Data were analysed using Student's t-test for paired data. We evaluated 44 centre residents (25 male, 19 female, average age 33 +/- 11 years). Mean lamotrigine dose was 272 +/- 133 mg per day. A significant reduction in seizure frequency was noted. Seizure frequency (all seizures) was 10.1 +/- 11.2 during the baseline period vs. 5.8 +/- 7.9 seizures per month during the treatment period (P = 0.002). Thirty-two percent of patients (n = 14) had greater than a 75% reduction in seizure frequency. Twenty-three percent of patients (n = 10) had a 50-74% seizure reduction. Twenty-five percent of patients (n = 11) had less than a 50% reduction in seizures, while 20% (n = 9) had an increase in seizures. A significant reduction of 48% in generalized seizures (9.5 +/- 11.6 vs. 4.9 +/- 6.5 seizures per month, P = 0.013) was noted. Reductions in partial seizure frequency of 48% (7.9 +/- 10 vs. 4 +/- 6.6 seizures per month, P = 0.16) as well as in mixed-type seizures (19.9 +/- 9.3 was vs. 15 +/- 12.1 seizures per month, P = 0.11) were also seen; however, these changes did not reach significance. Overall, lamotrigine was well tolerated by the subject population. Adjunctive treatment with lamotrigine appears to be an efficacious and well-tolerated treatment for seizures in a significant percentage of developmentally disabled patients with epilepsy. PMID- 10845739 TI - Efficacy and tolerability of topiramate in childhood and adolescent epilepsy: a clinical experience. AB - A 3-year retrospective review was undertaken of the use of topiramate in 51 children aged 3-16 years with partial and generalized epilepsies who attended a tertiary referral epilepsy centre in a large children's hospital. The mean follow up period was 19 months (range 6-33 months). Twenty-six children (51%) were still receiving topiramate at the time of their last review. Fifteen children (29%) showed a greater than 50% reduction in their seizure frequency and four children (8%) became seizure free, three on topiramate monotherapy. The drug appeared to be most effective in children with moderate learning difficulties with 75% showing an improvement in seizure control compared with 25% of children with normal educational functioning. Topiramate was withdrawn in 25 patients. The reasons for withdrawal included adverse effects in 20, lack of effect in three and worsening of seizures in two patients. Adverse side effects were reported in 57% of the 51 patients. The majority of the side effects were related to behavioural and cognitive difficulties, with less-common side effects including anorexia, weight loss and headaches. PMID- 10845740 TI - Closed head injury resulting in paradoxical improvement of a seizure disorder. AB - People with epilepsy are prone to head injuries and these traumas are usually considered adverse events. We report the case of a 31-year-old woman who sustained a closed head injury as a result of a seizure. This accidental trauma resulted in a 1-year remission of her previously intractable complex partial seizures. The paradoxical improvement of a seizure disorder can occur as a result of a closed head injury. We postulate it simulated ablative epilepsy surgery. PMID- 10845741 TI - Symptomatic generalized epilepsy associated with an inverted duplication of chromosome 15. AB - An inverted duplication of chromosome 15 (inv dup[15] chromosome) is the most common supernumerary marker chromosome in humans. Inv dup(15) chromosomes are commonly associated with mental retardation, epilepsy, behavioral problems and structural malformations. Though epilepsies associated with inv dup(15) chromosomes are often intractable, there have been very few reports regarding the seizure manifestations or types. We report a patient with severe mental retardation and intractable epilepsy, associated with an inv dup(15) chromosome. The seizures recorded with EEG-VTR monitoring were axial and generalized tonic seizures, and our case was diagnosed as symptomatic generalized epilepsy. Molecular and cytogenetic analysis showed an inv dup(15) chromosome containing the Prader-Willi syndrome/Angelman syndrome region mapped within bands 15q 11 q13. PMID- 10845742 TI - Prolonged post-ictal confusion as a manifestation of continuous complex partial status epilepticus: a depth EEG study. AB - We report a peculiar depth-EEG recording of prolonged post-ictal confusion which proved to be continuous complex partial status epilepticus. A 33 year old male with intractable medial temporal lobe epilepsy exhibited this ictal EEG recording. After repetitive habitual complex partial seizures, and an ensuing short lucid interval with intact memory and full communicability, the patient became more and more unresponsive and, finally, even cataleptic. Concurrent with this change in responsiveness, an EEG revealed a gradual and steady increase of ictal EEG activity. Immediately after intravenous diazepam infusion, this ictal EEG activity was suppressed and the patient began to move. This case confirms that a paradoxical excitation can occur after clustered complex partial seizures, instead of the well-known neuronal exhaustion. PMID- 10845743 TI - Estimates of smoking-attributable deaths at ages 15-54, motherless or fatherless youths, and resulting Social Security costs in the United States in 1994. AB - BACKGROUND: Deaths of parents often harm their children, taxpayers, and society, for decades. So we estimated the smoking-attributable (SA) counts and percentages (SA%) of U.S. 1994 deaths at child-rearing ages; youths (ages <18) left motherless or fatherless; and resulting Social Security Survivors Insurance taxes. DESIGN: U.S. 1994 age/sex/education-specific total and SA death counts were estimated using death certificate data and standard CDC SAMMEC methods (with added injury mortality), respectively. We separately summed (a) total and (b) SA age/sex/education-specific death counts times their average number of youths per adult (cumulative fertility, adjusted for infant mortality). We then multiplied the SA and total bereft youth counts by their average duration of Survivors Insurance, and calculated the SA cost of youth Survivors Insurance. RESULTS: In 1994, smoking caused an estimated 44,000 male and 19,000 female U.S. deaths at ages 15-54, leaving 31,000 fatherless and 12,000 motherless youths. On December 31, 1994, the SA prevalences [count (SA%)] of fatherless or motherless youths were an estimated 220,000 (17%) and 86,000 (16%), respectively. Resulting Survivors Insurance costs were about $1.4 (sensitivity range: $0.58-3.7) billion in 1994. CONCLUSIONS: Smoking causes many U.S. deaths at ages 15-54, youth bereavements, and Survivors Insurance costs. Reductions in smoking may greatly reduce those deaths, bereavements, and taxpayer and societal costs. PMID- 10845744 TI - Asthma. A message from Preventive Medicine and your physician. PMID- 10845745 TI - Determinants of infant feeding method in relation to risk factors for breast cancer. AB - BACKGROUND: Breastfeeding is considered to be an important factor for maternal and children's health. However, the epidemiological findings related to the effect of breastfeeding on women's health, especially with respect to breast cancer development, are inconsistent. Determinants of infant feeding method may contribute to the inconsistency. METHODS: A total of 24,769 women aged 40-64 in Miyagi Prefecture, Japan, responded to a self-administered questionnaire survey in 1990. Using the data obtained from 22,085 parous women, we calculated odds ratios (ORs) for the choice of "breastfeeding only" during reproductive period. RESULTS: Late age at menarche (> or = 16 years, OR = 1.57) and high body mass index (BMI) at 20 years of age (> or = 24, OR = 1.31) were associated with the choice of breastfeeding only. Late age at birth of first child (> or = 28 years, OR = 0.29), history of breast cancer in mother (OR = 0.68), and high educational level (more than a high school education, OR = 0.53) reduced the possibility of choosing breastfeeding only. CONCLUSION: The results indicate that the choice of infant feeding method is associated with several breast cancer risk factors. Based on this finding, we should construct appropriate breast cancer risk models for parous women and investigate the changes in the effects of breastfeeding and other breast cancer risk factors among these risk models. Especially in a risk model controlling for breastfeeding, the effects of other breast cancer risk factors should be reevaluated. Through comparisons among different risk models, we may find the best-fitted risk model and identify the true effect of breastfeeding. PMID- 10845746 TI - Changes in intensity of physical exercise as predictors of depressive symptoms among older adults: an eight-year follow-up. AB - BACKGROUND: Epidemiological research suggests that regular physical activity may be associated with reduced depressive symptoms. The present study examines the predictive value of physical exercise in relation to depressive symptoms among samples of adults aged 65+ during an 8-year period. METHODS: The subjects (N = 663) who participated both at the baseline (1988) and the follow-up (1996) interviews were selected for the analyses. The dependent variable depressive symptoms was assessed by the Finnish modified version of Beck's 13-item depression scale. The independent variable was the intensity of physical exercise. RESULTS: The intensity of physical exercise decreased among the older men and women. Those who had reduced their intensity of physical exercise during the 8 years reported more depressive symptoms at the follow-up than those who had remained active or increased their physical activity. Depressive symptoms were predicted by the intensity of baseline physical exercise, earlier depressive symptoms, older age, gender, having three or more chronic somatic conditions, and difficulties in performing ADL activities. CONCLUSIONS: Age-related decrease in the intensity of physical exercise increases the risk of depressive symptoms among older adults. This calls for effective measures in maintaining and supporting an adequate level of physical exercise among the aging population. PMID- 10845747 TI - Weight-control behaviors among adults and adolescents: associations with dietary intake. AB - BACKGROUND: This study aimed to assess the prevalence of weight-control behaviors and their associations with overall dietary intake among adults and adolescents. METHODS: Participants included 3,832 adults and 459 adolescents from four regions of the United States. Cross-sectional data were collected on energy and nutrient intake, weight-control behaviors, body mass index (BMI), and sociodemographics. RESULTS: Current weight-control behaviors were reported by 52.7% of the study population (adult women, 56.7%; adult men, 50.3%; adolescent girls, 44.0%; adolescent boys, 36.8%). Weight-control behaviors were consistently and positively associated with socioeconomic status among adults, but not among adolescents. Among "dieters," unhealthy practices were reported by 22.7% of adult women, 21.3% of adult men, 30.4% of adolescent girls, and 18.5% of adolescent boys. Adults trying to control their weight reported healthier nutrient intakes than those not trying to control their weight, in particular when moderate weight control methods were employed. Among adolescents, there were fewer differences across dieting status and these were not suggestive of healthier intakes among dieters than nondieters. CONCLUSIONS: Weight-control behaviors are reported by a large percentage of the population. Weight-control behaviors tend to be healthier among adults than among adolescents, in terms of the types of behaviors used and their impact on nutrient intakes. Obesity prevention interventions should emphasize the importance of using healthy weight-control practices. PMID- 10845748 TI - Characteristics of participants who stop smoking and sustain abstinence for 1 and 5 years in the Lung Health Study. AB - BACKGROUND: This study describes baseline and Year 1 predictors of abstinence from smoking for the 3,523 intervention participants who had complete annual 5 year follow-up data in the Lung Health Study (LHS). METHODS: The LHS enrolled 5,887 smokers, aged 35 to 60 years, of whom 3,923 were offered a cessation intervention. Of these, 22% achieved biochemically verified abstinence for 5 years. Logistic regressions were performed. The first outcome variable was abstinence from smoking at 1 year. Then for those who were quit at 1 year, the outcome variable was 5 years of sustained abstinence. RESULTS: All participants who were not using nicotine gum after 1 year in the study were more likely to sustain cessation over 5 years than were gum users at year 1 (OR ranged from 0.31 to 0.44 for four age- and sex-specific groups). Baseline number of previous quit attempts was negatively associated with 5-year quitting success among younger and older men (OR = 0.82 and 0.83). Older participants who were less likely to associate smoking with emotional coping had higher abstinence rates at 5 years of follow-up (OR = 0.89 and 0.84). CONCLUSIONS: Different mechanisms may be responsible for achieving cessation in age/gender groups. These results have implications for planning successful interventions. PMID- 10845749 TI - Age gradient in the cost-effectiveness of bicycle helmets. AB - OBJECTIVES: This study analyzed the reduction in risk of head injuries associated with use of bicycle helmets among persons ages 3 to 70 and the cost-effectiveness of helmet use based on this estimated risk reduction. METHODS: To derive our cost effectiveness estimates, we combined injury incidence data gathered through a detailed and comprehensive injury registration system in Norway, acute medical treatment cost information for the Norwegian health service, and information reported in the scientific literature regarding the health protective effects of helmet use. The analysis included all cases of head injuries reported through the registration system from 1990 through 1996. We performed an age-stratified analysis to determine the incidence of bicycle-related head injuries, the 5-year reduction in absolute risk of injury, the number needed to treat, and the cost effectiveness of helmet use. To test the robustness of the findings to parameter assumptions, we performed sensitivity analysis. RESULTS: The risk of head injury was highest among children aged 5 to 16. The greatest reduction in absolute risk of head injury, 1.0 to 1.4% over 5 years estimated helmet lifetime, occurred among children who started using a helmet between the ages of 3 and 13. Estimates indicate that it would cost approximately U.S. $2,200 in bicycle helmet expenses to prevent any one upper head injury in children ages 3-13. In contrast, it would cost U.S. $10,000-25,000 to avoid a single injury among adults. CONCLUSIONS: Bicycle safety helmets appear to be several times more cost-effective for children than adults, primarily because of the higher risk of head injury among children. Programs aiming to increase helmet use should consider the differences in injury risk and cost-effectiveness among different age groups and target their efforts accordingly. PMID- 10845750 TI - Health-promoting behaviors among adults with type 2 diabetes: findings from the Health and Retirement Study. AB - BACKGROUND: Type 2 diabetes has an enormous impact on the health care system and individuals. Dietary habits, exercise, weight management, and smoking status are critical to management and prevention of complications. This study describes the prevalence of these behaviors and their change over time in a national sample of adults with type 2 diabetes. The relationships between behavior change and sociodemographic and health status measures are explored. METHODS: Data are from the first and third longitudinal waves of the Health and Retirement Study. Surveys were conducted face-to-face or via telephone in 1992 and 1996. RESULTS: The sample consisted of 733 persons with type 2 diabetes, ages 50-62. The most common behaviors were being on a special diet (79.6%), and not smoking (76.6%). Sixty-six percent were engaged in some physical activity, and 58.4% were trying to lose weight. Reports of being on a special diet, trying to lose weight, and exercising all diminished over time. CONCLUSIONS: The prevalence of these behaviors is disappointing. Worse, they declined over the 4-year period. A better understanding of factors contributing to a person's decision to begin or discontinue health-promoting behaviors is needed to plan effective supportive or preemptive interventions. PMID- 10845751 TI - Differences in food patterns at breakfast by sociodemographic characteristics among a nationally representative sample of adults in the United States. AB - BACKGROUND: Eating breakfast is considered an important determinant of a healthy lifestyle. This study explores the different food patterns of breakfast for adults aged 18-65 in the United States. METHODS: Data are from the 1994-1996 Continuing Survey of Food Intake by Individuals, n = 15,641. Dietary assessment method used was the 24-h recall. Nutrient differences among the breakfast food patterns as well as the sociodemographic characteristics of individuals following each eating pattern are examined. RESULTS: The primary food patterns were based on consumption of eggs (15.3% of adults), ready-to-eat cereals (17.4%), bread (21.7%), cooked cereal (4.4%), fruit and fruit juice (5.5%), and coffee, soft drinks, and high-fat desserts (15.1%). Seventeen and three-tenths percent of the adults skipped breakfast. These food patterns provide remarkably different nutrient profiles adjusting for energy intake. The egg pattern is highest in total fat, lowest in fiber density, and low in iron and calcium density. In contrast, the ready-to-eat cereal pattern is high in fiber, highest in calcium density, and very low in fat. Breakfast food patterns differ markedly by various sociodemographic factors such as gender, ethnicity, and educational level. CONCLUSION: Different segments of our population consume different types of foods at breakfast, contributing to differences in their nutrient intakes. PMID- 10845752 TI - No evidence for multiple-drug prophylaxis for tuberculosis compared with isoniazid alone in Southeast Asian refugees and migrants: completion and compliance are major determinants of effectiveness. AB - BACKGROUND: The use of multiple-drug prophylaxis for tuberculosis (TB) has not been shown to be more effective than prophylaxis with isoniazid alone. The boundary between inactive pulmonary TB (class 4 TB) and culture-negative "active" pulmonary TB (class 3 TB) is often unclear, as is the intention to treat such patients as a preventive measure or as a curative measure. METHODS: We compared the effectiveness of single drug preventive therapy with isoniazid to the effectiveness of multiple drug preventive therapy for patients with asymptomatic, inactive TB, in a retrospective cohort study of 984 Southeast (SE) Asian migrants and refugees who received prophylaxis between 1978 and 1980. RESULTS: The rate of TB developing in this cohort was 122 per 100,000 person-years. There was no significant difference in development of TB between people who received isoniazid only and those who received multiple drugs. The only significant predictor of TB was noncompletion of prophylaxis [relative risk (RR) = 62, 95% confidence interval (CI) = 20-194]. Subgroup analysis on people who had completed therapy showed noncompliance as a significant predictor of TB (RR = 16, 95% CI = 1.4 179). The risk of noncompletion (RR = 4.7, 95% CI = 2.37-9.39, P < 0.0001) and noncompliance (RR = 2.2, 95% CI = 1.03-4.7, P = 0.03) was higher for patients who received multiple drugs compared with isoniazid alone. Multiple-drug therapy cost 30 times more than isoniazid alone. CONCLUSIONS: We did not find evidence in support of the empirical practice of giving multiple drugs for prevention of TB. This practice is also more costly and more likely to result in noncompliance and adverse drug reactions. PMID- 10845753 TI - Sociocultural variables in youth access to tobacco: replication 5 years later. AB - BACKGROUND: A prior study presented the only systematic investigation of the role of sociocultural variables in youth access to tobacco. White, black, and Latino girls and boys attempted to purchase cigarettes in the same 72 stores at the same time of day. Results revealed significantly greater sales to girls than to boys and to minorities than to whites. Before concluding that sociocultural variables must be addressed in merchant intervention programs designed to reduce youth access to tobacco, this study must be replicated, particularly in light of the significant decreases in youth access in the past 5 years. This article presents that replication. METHOD: The stores used in the prior study were selected, and 12 white, black, and Latino girls and boys attempted to purchase cigarettes in those stores at the same time of day. Results Youths' access rate in 1999 (12.7%) was significantly lower than in the prior (1993-1995) study (41%). No effect for minors' gender was found, but the ethnicity effect again emerged: Black and Latino youth were 2.5 times more likely to be sold cigarettes than their white counterparts. CONCLUSIONS: Multiple sociocultural variables affect youth access to tobacco when access rates are high, but only youth ethnicity plays a role when access rates are low. Merchant interventions designed to reduce youth access to tobacco must address ethnic issues. PMID- 10845754 TI - What does glycated hemoglobin measure? PMID- 10845755 TI - Respiratory network function in the isolated brainstem-spinal cord of newborn rats. AB - The in vitro brainstem-spinal cord preparation of newborn rats is an established model for the analysis of respiratory network functions. Respiratory activity is generated by interneurons, bilaterally distributed in the ventrolateral medulla. In particular non-NMDA type glutamate receptors constitute excitatory synaptic connectivity between respiratory neurons. Respiratory activity is modulated by a diversity of neuroactive substances such as serotonin, adenosine or norepinephrine. Cl(-)-mediated IPSPs provide a characteristic pattern of membrane potential fluctuations and elevation of the interstitial concentration of (endogenous) GABA or glycine leads to hyperpolarisation-related suppression of respiratory activity. Respiratory rhythm is not blocked upon inhibition of IPSPs with bicuculline, strychnine and saclofen. This indicates that GABA- and glycine mediated mutual synaptic inhibition is not crucial for in vitro respiratory activity. The primary oscillatory activity is generated by neurons of a respiratory rhythm generator. In these cells, a set of intrinsic conductances such as P-type Ca2+ channels, persistent Na+ channels and G(i/o) protein-coupled K+ conductances mediates conditional bursting. The respiratory rhythm generator shapes the activity of an inspiratory pattern generator that provides the motor output recorded from cranial and spinal nerve rootlets in the preparation. Burst activity appears to be maintained by an excitatory drive due to tonic synaptic activity in concert with chemostimulation by H+. Evoked anoxia leads to a sustained decrease of respiratory frequency, related to K+ channel-mediated hyperpolarisation, whereas opiates or prostaglandins cause longlasting apnea due to a fall of cellular cAMP. The latter observations show that this in vitro model is also suited for analysis of clinically relevant disturbances of respiratory network function. PMID- 10845756 TI - Astrocyte line SVG-TH grafted in a rat model of Parkinson's disease. AB - The present review describes gene transfer into the brain using extraneuronal cells with an ex vivo approach. The mild immunological reactions in the central nervous system to grafts provided the rationale and empirical basis for brain transplantation, to replace dying cells, of potential clinical relevance. Fetal human astrocytes were genetically engineered to express tyrosine hydroxylase, the rate-limiting enzyme for the synthesis of catecholamines. These cells were also found to produce constitutively and secrete GDNF and interleukins. Therefore, these cells may prove as a drug-delivery system for the treatment of neurological degenerative conditions such as Parkinson's disease (PD). The field of neuronal reconstruction has reached a critical threshold and there is a need to evaluate the variables that will become critical as the field matures. One of the needs is to characterize the neurochemical alterations in the microenvironment in the context of grafted-host connectivity. This review discusses the functional effects of the pharmacologically-active construct, which consists of astrocytes producing L-DOPA and GDNF. The striatum in PD that lacks the dopaminergic projection from the substantia nigra metabolizes and releases dopamine differently from normal tissue and may react to different factors released by the grafted cells. Moreover, neurochemicals of the host tissue may effect grafted cells as well. An understanding of the way in which these neurochemicals are abnormal in PD and their role in the grafted brain is critical to the improvement of reconstructive strategies using cellular therapeutic strategies. PMID- 10845757 TI - Trophic effects of purines in neurons and glial cells. AB - In addition to their well known roles within cells, purine nucleotides such as adenosine 5' triphosphate (ATP) and guanosine 5' triphosphate (GTP), nucleosides such as adenosine and guanosine and bases, such as adenine and guanine and their metabolic products xanthine and hypoxanthine are released into the extracellular space where they act as intercellular signaling molecules. In the nervous system they mediate both immediate effects, such as neurotransmission, and trophic effects which induce changes in cell metabolism, structure and function and therefore have a longer time course. Some trophic effects of purines are mediated via purinergic cell surface receptors, whereas others require uptake of purines by the target cells. Purine nucleosides and nucleotides, especially guanosine, ATP and GTP stimulate incorporation of [3H]thymidine into DNA of astrocytes and microglia and concomitant mitosis in vitro. High concentrations of adenosine also induce apoptosis, through both activation of cell-surface A3 receptors and through a mechanism requiring uptake into the cells. Extracellular purines also stimulate the synthesis and release of protein trophic factors by astrocytes, including bFGF (basic fibroblast growth factor), nerve growth factor (NGF), neurotrophin-3, ciliary neurotrophic factor and S-100beta protein. In vivo infusion into brain of adenosine analogs stimulates reactive gliosis. Purine nucleosides and nucleotides also stimulate the differentiation and process outgrowth from various neurons including primary cultures of hippocampal neurons and pheochromocytoma cells. A tonic release of ATP from neurons, its hydrolysis by ecto-nucleotidases and subsequent re-uptake by axons appears crucial for normal axonal growth. Guanosine and GTP, through apparently different mechanisms, are also potent stimulators of axonal growth in vitro. In vivo the extracellular concentration of purines depends on a balance between the release of purines from cells and their re-uptake and extracellular metabolism. Purine nucleosides and nucleotides are released from neurons by exocytosis and from both neurons and glia by non-exocytotic mechanisms. Nucleosides are principally released through the equilibratory nucleoside transmembrane transporters whereas nucleotides may be transported through the ATP binding cassette family of proteins, including the multidrug resistance protein. The extracellular purine nucleotides are rapidly metabolized by ectonucleotidases. Adenosine is deaminated by adenosine deaminase (ADA) and guanosine is converted to guanine and deaminated by guanase. Nucleosides are also removed from the extracellular space into neurons and glia by transporter systems. Large quantities of purines, particularly guanosine and, to a lesser extent adenosine, are released extracellularly following ischemia or trauma. Thus purines are likely to exert trophic effects in vivo following trauma. The extracellular purine nucleotide GTP enhances the tonic release of adenine nucleotides, whereas the nucleoside guanosine stimulates tonic release of adenosine and its metabolic products. The trophic effects of guanosine and GTP may depend on this process. Guanosine is likely to be an important trophic effector in vivo because high concentrations remain extracellularly for up to a week after focal brain injury. Purine derivatives are now in clinical trials in humans as memory-enhancing agents in Alzheimer's disease. Two of these, propentofylline and AIT-082, are trophic effectors in animals, increasing production of neurotrophic factors in brain and spinal cord. Likely more clinical uses for purine derivatives will be found; purines interact at the level of signal-transduction pathways with other transmitters, for example, glutamate. They can beneficially modify the actions of these other transmitters. PMID- 10845758 TI - The selective vulnerability of striatopallidal neurons. AB - The different types of striatal neuron show a range of vulnerabilities to a variety of insults. This can be clearly seen in Huntington's disease where a well mapped pattern of pathological events occurs. Medium spiny projection (MSP) neurons are the first striatal cells to be affected as the disease progresses whilst interneurons, in particular the NADPH diaphorase positive ones, are spared even in the late stages of the disease. The MSP neurons themselves are also differentially affected. The death of MSP neurons in the patch compartment of the striatum precedes that in the matrix compartment and the MSP neurons of the dorsomedial caudate nucleus degenerate before those in the ventral lateral putamen. The enkephalin positive striatopallidal MSP neurons are also more vulnerable than the substance P/dynorphin MSP neurons. We review the potential causes of this selective vulnerability of striatopallidal neurons and discuss the roles of endogenous glutamate, nitric oxide and calcium binding proteins. It is concluded that MSP neurons in general are especially susceptible to disruptions of cellular respiration due to the enormous amount of energy they expend on maintaining unusually high transmembrane potentials. We go on to consider a subpopulation of enkephalinergic striatopallidal neurons in the rat which are particularly vulnerable. This subpopulation of neurons readily undergo apoptosis in response to experimental manipulations which affect dopamine and/or corticosteroid levels. We speculate that the cellular mechanisms underlying this cell death may also operate in degenerative disorders such as Huntington's disease thereby imposing an additional level of selectivity on the pattern of degeneration. The possible contribution of the selective death of striatopallidal neurons to a number of clinically important psychiatric conditions including obsessive compulsive disorders and Tourette's syndrome is also discussed. PMID- 10845759 TI - Sympathetic pathways and adrenergic innervation of the penis. AB - The sympathetic nervous system is important for penile function: it mediates detumescence and may contribute to the maintenance of the penis in a non-erect state. The sympathetic preganglionic neurons are found in the intermediolateral gray matter of the spinal cord. Postganglionic neurons are located to the sympathetic chain ganglia, the inferior mesenteric, hypogastric and pelvic ganglia, and possibly to ganglia near the target organ. Sympathetic fibres can be found in the pelvic, cavernous, and pudendal nerves. Stimulation of the sympathetic pathways to the penis may, however, also produce erection. It has been suggested that the suprasacral vasodilator pathway is a sympathetic cholinergic pathway, operating through cholinergic neurons in the pelvic plexus. In the penis, there is a rich sympathetic, adrenergic innervation of the corpus cavernosum (CC) and the vasculature, and in particular of the helicine arteries. Sympathetic, adrenergic nerves also contain neuropeptide Y. Parasympathetic cholinergic nerves, which mediate CC relaxation and erection, contain not only acetylcholine, but also vasoactive intestinal polypeptide, nitric oxide synthase, and probably other mediators and/or mediator-synthesizing enzymes. Activation of sympathetic adrenergic nerves causes release of noradrenaline, acting on alpha adrenoceptors in the trabecular smooth muscle of the CC and in penile vessels. The role of interactions between different transmitters and mediators, released from nerves or generated locally, in the regulation of contraction and relaxation of CC and penile vessels, needs further study. PMID- 10845760 TI - Central noradrenergic control of penile erection. AB - Penile erection is completely dependent on commands from the central nervous system. Spinal centers controlling penile erection are located in the thoracolumbar and lumbosacral spinal cord. These centers are activated by information from the periphery and supraspinal nuclei so as to elicit penile erection in a variety of physiological contexts. A small number of nuclei including the locus coeruleus located in the pons sends noradrenergic fibers to the forebrain and spinal cord, including those areas controlling penile erection. Recent morphological techniques such as in situ hybridization and autoradiography using radioligand binding permit investigation of the brain and spinal pathways utilizing alpha adrenoceptor subtypes. Furthermore, pharmacological experiments suggest a modulatory role for noradrenaline in the control of penile erection either in the brain or in the spinal cord. The most robust evidence is that central inhibition of alpha-2 adrenoceptors facilitates sexual function. Taken together, the data propose new directions in the physiological exploration of penile erection and the therapeutic approach of erectile dysfunction. PMID- 10845761 TI - Regulation of pre-synaptic alpha adrenergic activity in the corpus cavernosum. AB - The neurotransmitters and vasoactive substances regulating tone in the smooth muscle of the penile arteries/arterioles and the trabeculae of the corpora cavernosa are critical mediators of the state of penile erection. Contemporary research reveals a coordinated, intricate interplay between the pathways of vasorelaxation and vasoconstriction representing a most efficient physiological mechanism to initiate and maintain penile erection. This paper will focus on the role of the adrenergic constrictor pathways in penile erection and, more specifically, on the pre-junctional adrenergic mechanisms that regulate smooth muscle constriction. All neurogenic constrictor responses are related to the release of norepinephrine from adrenergic nerves that act on post-junctional alpha-1 and pre-junctional and post-junctional alpha-2 receptor subtypes. Based on the current state of knowledge, there are at least three pre-junctional mechanisms regulating penile smooth muscle tone. First, norepinephrine release from the adrenergic nerves binds to the pre-junctional alpha-2 adrenoceptor on the adrenergic nerves and negatively regulates norepinephrine release. Blockade of this reaction by selective alpha-2 receptor antagonists (e.g. yohimbine or delequamine) will enhance norepinephrine release. Second, norepinephrine release from the adrenergic nerves binds to the pre-junctional alpha-2 adrenoceptor on the non-adrenergic, non-cholinergic (NANC) nerves and inhibits nitric oxide synthesis and release. Blockade of this reaction by selective alpha-2 receptor antagonists (e.g. yohimbine or delequamine) will enhance nitric oxide release, facilitating erection. Finally, cholinergic nerves pre-junctionally inhibit norepinephrine release from the adrenergic nerve and stimulate the NANC nerve to increase nitric oxide synthesis and release. These observations indicate that different neurotransmitters regulate the adrenergic neurotransmission pathway. Based on the above concepts for pre-junctional and post-junctional regulation of smooth muscle tone, the most efficacious strategy to reduce adrenergic activity and facilitate penile erection is to combine alpha-1 and alpha-2 adrenergic receptor antagonists. In this case, any enhancement of norepinephrine release is of little importance because the alpha-1 receptor antagonist will impede this vasoconstrictor response. This will also enhance the release of nitric oxide, which increases smooth muscle relaxation and decreases contraction resulting in penile erection. PMID- 10845762 TI - Alpha-adrenergic receptor blockade by phentolamine increases the efficacy of vasodilators in penile corpus cavernosum. AB - Penile trabecular smooth muscle tone, a major determinant of erectile function, is highly regulated by numerous inter- and intracellular pathways. The interaction between pathways mediating contraction and relaxation has not been studied in detail. To this end, we investigated the functional effects of alpha adrenergic receptor blockade with phentolamine and its interaction with vasodilators (sildenafil, vasoactive intestinal polypeptide (VIP) and PGE1) that elevate cyclic nucleotides on penile cavernosal smooth muscle contractility. In organ bath preparations of cavernosal tissue strips contracted with phenylephrine, phentolamine significantly enhanced relaxation induced by sildenafil, VIP and PGE1. Sildenafil, VIP or PGE1 also significantly enhanced relaxation induced by phentolamine in cavernosal tissue strips contracted with phenylephrine. To study the effects of alpha adrenergic receptor blockade and modification of cyclic nucleotide metabolism during active neurogenic input, cavernosal tissue strips in organ bath preparations were contracted with the non adrenergic agonist endothelin-1 and subjected to electrical field stimulation (EFS) in the absence or presence of phentolamine and/or sildenafil. EFS (5-40Hz) typically caused biphasic relaxation and contraction responses. Phentolamine alone enhanced relaxation and reduced or prevented contraction to EFS. Sildenafil enhanced relaxation to EFS at lower frequencies (< or = 5 Hz). The combination of phentolamine and sildenafil enhanced EFS-induced relaxation at all frequencies tested. EFS, in the presence of 10 nM phentolamine and 30 nM sildenafil, produced enhanced relaxation responses which were quantitatively similar to those obtained in the presence of 50 nM sildenafil alone. Thus, blockade of alpha-adrenergic receptors with phentolamine increases the efficacy of cyclic nucleotide-dependent vasodilators. Furthermore, phentolamine potentiates relaxation and attenuates contraction in response to endogenous neurotransmitters which are released during EFS. These findings suggest that antagonism of alpha-adrenergic signaling enables other independent relaxatory pathways to predominate within penile trabecular smooth muscle. PMID- 10845763 TI - Abanoquil, a new alpha-1 adrenoceptor antagonist. In vitro and in vivo effect on erectile tissue. AB - Using an organ bath model with porcine cavernosal tissue strips and an in vivo monkey model we demonstrated that abanoquil, a novel alpha adrenoceptor antagonist, is able to relax contracted tissue strips and induce erectile response when injected intracorporally. The erectile response in the monkeys was not dose-related and compared to the effect of papaverine injections, abanoquil induced a lower level of tumescence and rigidity. Hence, abanoquil might be useful as a facilitator of erection in the pharmacological treatment of erectile dysfunction. PMID- 10845764 TI - Cyclic AMP regulates mRNA expression of alpha-1d and alpha-2a adrenergic receptors in cultured human corpus cavernosum smooth muscle cells. AB - While the physiological effects of contractile (e.g. norepinephrine) and relaxatory (e.g. PGE1, forskolin) agents on corpus cavernosum smooth muscle tone have been characterized, the regulation of alpha adrenergic receptor mRNA expression in erectile tissue remains to be investigated. The goal of this study was to investigate the modulation of alpha-1 and alpha-2 adrenergic receptor mRNA expression in cultured human corpus cavernosum smooth muscle cells in response to increased intracellular cAMP induced by prostaglandin E1 and forskolin. Human corpus cavernosum smooth muscle cells were incubated for 24 h with or without PGE1 (5.7 microM), forskolin (10 microM) or an admixture of both. Total RNA was prepared from the cultures. Expression of alpha-1d adrenergic receptor, alpha-2a adrenergic receptor and m2 muscarinic acetylcholine receptor was determined by RNase protection assays. Loading was normalized by RNase protection of the housekeeping gene, cyclophilin A. The relative abundance of mRNAs was quantitated by scanning densitometry. Treatment of human corpus cavernosum smooth muscle cells with PGE1 or forskolin resulted in decreased mRNA expression of alpha-1d and alpha-2a adrenergic receptors and m2 muscarinic acetylcholine receptor when compared to untreated cells. Combinations of PGE1 and forskolin produced a more pronounced decrease in mRNA than either agent alone. PGE1 and forskolin increased intracellular levels of cAMP in human corpus cavernosum smooth muscle cells and combinations of both agents produced a more pronounced increase in cAMP synthesis. These results suggest that cAMP modulates the expression of alpha adrenergic receptors, one of the principal contractile receptor systems in the corpora cavernosa. These observations further support the concept that erectile function is a balance between contractile and relaxatory processes, which in turn regulate structure and function of the corpora cavernosa. PMID- 10845765 TI - Role of alpha adrenergic receptors in erectile function. AB - Penile erection is a complex physiological process in which the regulation of penile hemodynamics depends on signal input from central and peripheral nervous systems, and on the balance and interplay between several local physiological mediators (neurotransmitters, vasoactive agents and endocrine factors). A role for the sympathetic nervous system in attaining or maintaining penile flaccidity and detumescence is well recognized. However, the exact mechanisms of alpha adrenergic receptor mediated erectile function remain poorly defined. The objective of this review is to summarize data presented in the literature and from our laboratory on alpha-adrenergic receptors, and to discuss the conceptual framework by which the alpha-adrenergic receptor pathway modulates penile corpus cavernosum smooth muscle contractility. We will integrate the current state of knowledge of penile erection into one framework encompassing the biochemical and physiological pathways responsible for trabecular smooth muscle relaxation (erection) and contraction (detumescence). We will focus our discussion on local control mechanisms of alpha-adrenergic receptors and explore the following topics: (1) functional activity of alpha-1 and alpha-2 adrenergic receptors in the physiology of human penile erection; (2) identification, classification and characterization of alpha-1 and alpha-2 adrenergic receptor subtypes in human penile erectile tissue; (3) molecular mechanism of action of alpha-1 and alpha-2 adrenergic receptors in human penile erectile tissue; (4) blockade of alpha-1 and alpha-2 adrenergic receptor action by selective and non-selective alpha-1 and alpha-2 adrenergic receptor antagonists (blockers); (5) physiologic (functional) antagonism of alpha-1 and alpha-2 adrenergic receptor activity by vasodilators mediating smooth muscle relaxation; and (6) key areas of consensus, as well as critical gaps in the existing scientific knowledge concerning the rationale and the potential use of alpha-blockers in erectile function. PMID- 10845766 TI - Effects of alpha-2 blockade on sexual response: experimental studies with delequamine (RS15385). AB - The role of alpha-2 receptors and alpha-2 antagonists in central and peripheral mechanisms of sexual response are discussed. It is concluded that the predominant role of the alpha-2 antagonist centrally is to increase arousal which in certain circumstances, is sexual. It is further concluded that the predominant role of the alpha-2 antagonist peripherally is to modulate (block) the norepinephrine (NE)-induced contractility in the smooth muscle of the penis. How the central arousal mechanisms are specifically linked to sexual response is not understood. Experimental studies with a selective alpha-2 antagonist, delequamine, are briefly reviewed and their complex results discussed. Evidence from sleep studies was consistent with delequamine having both central excitatory and inhibitory effects, dependent on dosage. The possibility that men with psychogenic erectile dysfunction might have increased central alpha-2 tone was considered. The apparent loss of responsiveness to the alpha-2 antagonist in older dysfunctional men was discussed. More questions are raised than answered; further research is needed in this area. PMID- 10845767 TI - Yohimbine in erectile dysfunction: the facts. AB - Yohimbine, a pharmacologically well-characterized alpha-2-adrenoceptor antagonist with activity in the central and peripheral nervous system, has been used for over a century in the treatment of erectile dysfunction. In-depth, systematic studies in animals have shown that the drug has a remarkable positive effect on sexual performance. Meta-analyses of the few controlled, randomized human studies have consistently shown an advantage of yohimbine over placebo. Despite such a long history and encouraging activity, the drug has not yet been subjected to scientifically rigorous human clinical trials. Although relevant basic pharmacological and animal research information has been available for over 15 y, recent studies were designed with a lack of insight and complete disregard of those fundamental studies. Currently, dose-response investigations are not available, alternative routes of administration (i.e. sublingual) have not been investigated, nor has continuous versus 'on-demand' administration been explored. Synergistic activity with other drugs was last studied nearly four decades ago. Assessment of various populations was carried out in very limited cohorts and only in most general terms. In short, properly designed trials in humans have not been done. Why? Yohimbine is an old drug. As such it does not enjoy patent protection or commercial viability. Until molecular/formulation changes can be brought about (as recently happened with two other agents: phentolamine and apomorphine), serious investigations of yohimbine will remain in limbo. It could be that the nay sayers are right and yohimbine, indeed, lacks clinical activity as a treatment for men with erectile dysfunction. As long as it remains an orphan drug, we will never know. PMID- 10845768 TI - Oral phentolamine: an alpha-1, alpha-2 adrenergic antagonist for the treatment of erectile dysfunction. AB - Phentolamine mesylate is an alpha-1 and alpha-2 selective adrenergic receptor antagonist which has undergone clinical trials for erectile dysfunction treatment. Biochemical and physiological studies in human erectile tissue have revealed a high affinity of phentolamine for alpha-1 and alpha-2 adrenergic receptors. Based on pharmacokinetic studies, it is suggested that 30-40 min following oral ingestion of 40 or 80 mg of phentolamine (Vasomax), the mean plasma phentolamine concentrations are sufficient to occupy the alpha-1 and -2 adrenergic receptors in erectile tissue and thereby result in inhibition of adrenergic-mediated physiologic activity. In large multi-center, placebo controlled pivotal phase III clinical trials, the mean change in the erectile function domain of the International Index of Erectile Function scores (Questions 1-5 and 15) from screening to the end of treatment was significantly higher following use of active drug (40 mg and 80 mg) compared to placebo. Three to four times as many patients receiving phentolamine reported being satisfied or very satisfied compared with those receiving placebo. At doses of 40 mg and 80 mg respectively, 55% and 59% of men were able to achieve vaginal penetration with 51% and 53% achieving penetration on 75% of attempts. The correction of erectile dysfunction or improvement to a less severe category of dysfunction was experienced by 53% of men with the 80 mg dose and 40% with the 40 mg dose of phentolamine. All trends of response were the same regardless of any concomitant medication. There were no severe adverse events. At 40 mg, 7.7% experienced rhinitis and fewer than 3.1% experienced any other side effect of treatment. Phentolamine is safe, well tolerated and efficacious for the treatment of erectile dysfunction. PMID- 10845769 TI - Comparison of oral and intracavernosal vasoactive agents in penile erection. AB - This discussion summarizes the potential differences in physiologically and pharmacologically induced erections and highlights the possible differences in the pathways facilitated by oral versus intracavernosal agents in penile erection. Oral agents act in conjunction with sexual stimulation either increasing corporal smooth muscle relaxation or attenuating smooth muscle contraction. However, their efficacy is dependent on sexual stimulation. Intracavernosal administration of phosphodiesterase type 5 inhibitor (sildenafil) or short-acting alpha adrenergic receptor antagonist (phentolamine), in the absence of sexual stimulation, does not initiate penile erection. In contrast, intracavernosal administration of PGE1 or papaverine induces erection independent of sexual stimulation. Thus, oral agents are not direct mediators of smooth muscle relaxation and act to facilitate relaxation in response to sexual stimulation, while intracavernosal agents directly mediate smooth muscle relaxation, independent of sexual stimulation. Although, considerable advances have been made in elucidating the physiology and pharmacology of erectile function, details of the signal transduction pathways affected by these agents in the penile corpus cavernosum are yet to be fully investigated. PMID- 10845770 TI - Diversity of expression of the sensory neuron-specific TTX-resistant voltage gated sodium ion channels SNS and SNS2. AB - The differential distribution of two tetrodotoxin resistant (TTXr) voltage-gated sodium channels SNS (PN3) and SNS2 (NaN) in rat primary sensory neurons has been investigated. Both channels are sensory neuron specific with SNS2 restricted entirely to those small dorsal root ganglion (DRG) cells with unmyelinated axons (C-fibers). SNS, in contrast, is expressed both in small C-fiber DRG cells and in 10% of cells with myelinated axons (A-fibers). All SNS expressing A-fiber cells are Trk-A positive and many express the vanilloid-like receptor VRL1. About half of C-fiber DRG neurons express either SNS or SNS2, and in most, the channels are colocalized. SNS and SNS2 are found both in NGF-responsive and GDNF-responsive C fibers and many of these cells also express the capsaicin receptor VR1. A very small proportion of small DRG cells express either only SNS or only SNS2. At least four different classes of A- and C-fiber DRG neurons exist, therefore, with respect to expression of these sodium channels. PMID- 10845771 TI - Myelin basic protein gene dosage effects in the PNS. AB - Myelin basic protein (MBP) plays an essential adhesive role in the formation of compact myelin in the central nervous system (CNS), but not in the peripheral nervous system (PNS). Morphologic data suggest that MBP controls the number of cytoplasmic channels or Schmidt-Lanterman incisures (SLI) present in PNS myelin. The levels of connexin-32 (Cx32) and myelin-associated glycoprotein (MAG), two components of the incisures, are inversely proportional to the levels of MBP in sciatic nerves of mice affected by the shiverer (shi) mutation, while protein zero (P0) and peripheral membrane protein 22 (PMP22), two structural components of compact myelin, remain constant. The levels of P0, PMP22, Cx32, and MAG mRNA do not vary in relationship to the levels of MBP. This indicates that MBP exerts its effect on Cx32 and MAG at a posttranscriptional level and suggests a new function for MBP in regulating gene expression in the PNS. PMID- 10845772 TI - Developmental regulation of amyloid precursor protein at the neuromuscular junction in mouse skeletal muscle. AB - Amyloid precursor protein (APP), associated with Alzheimer's disease plaques, is known to be present in synapses of the brain and in the adult neuromuscular junction (NMJ). In the present study we examined protein and gene expression of APP during the development of mouse skeletal muscle. Using immunocytochemical approaches, we found that APP is first detected in myotube cytoplasm at embryonic day 16 and becomes progressively concentrated at the NMJ beginning at birth until adulthood. The colocalization between APP and acetylcholine receptors at the NMJ is only partial at birth, but becomes complete upon reaching adulthood. We observed that all APP isoforms, including the Kunitz-containing (protease inhibitor or KPI) forms, are up-regulated from birth to postnatal day 5 and then decreased to reach the low levels observed in the adult. This suggests the involvement of APP during the events which lead to a mature mono-innervated synapse. A 92-kDa band, characteristic of a cleaved APP695 isoform and not due to a new muscle-specific alternative spliced form, was observed from postnatal day 15 following completion of polyneuronal synapse elimination. Taken together, these data suggest that skeletal muscle APP may well play a role in the differentiation of skeletal muscle and in the formation and maturation of NMJs. PMID- 10845773 TI - Involvement of caspase 3 in apoptotic death of cortical neurons evoked by DNA damage. AB - Previous reports have shown that DNA-damage-evoked death of embryonic cortical neurons is delayed by general caspase inhibitors and is accompanied by an increase in DEVD-AFC cleavage activity. We show here that this cleavage activity is lacking in camptothecin-treated caspase 3-deficient neurons. Moreover, we report that death of camptothecin-treated caspase 3-deficient neurons cultured from E16 embryos is delayed and that no significant increase in survival is observed with cotreatment with the general caspase inhibitor BAF. These results indicate that caspase-dependent death of camptothecin-treated cortical neurons requires caspase 3 activity. The delay in death is accompanied by impairment of DNA fragmentation. However, Bax-dependent cytochrome c release still occurs in camptothecin-treated caspase 3-deficient cortical neurons. Accordingly, we hypothesize that the delayed death which occurs in the absence of caspase 3 activity may be due to mitochondrial dysfunction. Finally, we show that the delay in death observed with E16 caspase 3-deficient neurons does not occur in neurons cultured from E19 embryos. This suggests that the requirement for caspase 3 in death of neurons evoked by DNA damage may differ depending upon the developmental state of the cell. PMID- 10845774 TI - N-acetyllactosamine and the CT carbohydrate antigen mediate agrin-dependent activation of MuSK and acetylcholine receptor clustering in skeletal muscle. AB - Galbeta1,3GalNAc and Galbeta1,4GIcNAc are the subterminal saccharide structures present on the CT carbohydrate antigen GalNAcbeta1,4[NeuAcalpha2,3]-Galbeta1 (3GalNAc or 4GIcNAc)-R, which is localized at the mammalian neuromuscular junction. Here we show that Galbeta1,3GalNAc, Galbeta1,4GIcNAc, and the CT carbohydrate antigen affect postsynaptic assembly in cultured muscle cells. Treatment of C2C12 myotubes with benzyl-O-alpha-GalNAc or neuraminidase increased peanut agglutinin (PNA) expression and AChR clustering. Induction of AChR clustering was blocked by PNA and by muscle agrin. Addition of Galbeta1,4GIcNAc or Galbeta1,3GalNAc increased AChR clustering in myotubes and muscle-specific kinase (MUSK) autophosphorylation in vitro, while NeuAcalpha2,3Galbeta1,4GIcNAc and Galbeta1,4GIc did not. Neural agrin activated MuSK in vitro if the lactosamine-containing mucin domain was present, and this activation was blocked in large part by Galbeta1,3GalNAc and Galbeta1,4GIcNAc. Agrin fragments and MuSK bound to these disaccharides with differing specificities. Overexpression of the CT carbohydrate antigen also increased AChR clustering and MuSK autophosphorylation in the presence of neural agrin. These data suggest a model in which different portions of the CT carbohydrate structure contribute to agrin dependent signal transduction. PMID- 10845775 TI - Neurofibromin negatively regulates neurotrophin signaling through p21ras in embryonic sensory neurons. AB - Embryonic sensory and sympathetic neurons that lack neurofibromin, the protein product of the neurofibromatosis type 1 (Nfl) gene, survive and extend neurites in the absence of neurotrophins. To determine whether neurofibromin negatively regulates neurotrophin signaling through its interaction with p21ras, we used Fab antibody fragments to block Ras function in DRG, trigeminal, nodose, and SCG neurons isolated from Nfl(-/-) and wild-type mouse embryos. We show that introduction of anti-Ras Fab fragments significantly reduces the ability of neurofibromin-deficient neurons to survive in the absence of neurotrophins. Moreover, addition of H-ras protein enhances the survival of Nfl(-/-), but not wild-type, DRG neurons. Our results are consistent with a major role for neurofibromin in modulating Trk signaling through p21ras during neuronal development. PMID- 10845776 TI - Synaptobrevin 2 is palmitoylated in synaptic vesicles prepared from adult, but not from embryonic brain. AB - Neuronal SNARE-proteins such as synaptobrevin, SNAP 25, and synaptotagmin are key players during neurosecretion. So far palmitoylation of SNAP-25 and synaptotagmin 1 have been described in vivo. Here we have analyzed palmitoylation of the SNARE proteins synaptobrevin 2 and synaptotagmin in vitro using synaptosomal and synaptic vesicle preparations from rat brain. Labeling of synaptic vesicles prepared from adult brain with [3H]palmitate revealed synaptobrevin 2 besides synaptotagmin 1 as major palmitoylated proteins. [3H]Palmitoylation of synaptobrevin 2 was resistant to chloroform/methanol extraction, but sensitive to reducing agents indicating a covalent fatty acid bond to cysteine residues. Palmitoylation of synaptobrevin 2 was also confirmed using endogenous synaptobrevin 2 present in PC-12 cells and synaptobrevin 2 expressed with a vacciniavirus system in Cos cells. In contrast to the situation seen with membrane preparations obtained from adult brain, synaptic vesicles prepared from embryonic rat brain did not support [3H]palmitoylation of synaptobrevin and synaptotagmin. These results suggest, that both synaptobrevin 2 and synaptotagmin were efficiently palmitoylated from mature synaptic vesicles. However, at least one component of the palmitoylation machinery is developmentally upregulated. PMID- 10845777 TI - How lead exposure relates to temporal changes in IQ, violent crime, and unwed pregnancy. AB - This study compares changes in children's blood lead levels in the United States with subsequent changes in IQ, based on norm comparisons for the Cognitive Abilities Test (CogAT) given to representative national samples of children in 1984 and 1992. The CogAT norm comparisons indicate shifts in IQ levels consistent with the blood lead to IQ relationship reported by an earlier study and population shifts in average blood lead for children under age 6 between 1976 and 1991. The CogAT norm comparisons also support studies indicating that the IQ to blood lead slope may increase at lower blood lead levels. Furthermore, long-term trends in population exposure to gasoline lead were found to be remarkably consistent with subsequent changes in violent crime and unwed pregnancy. Long term trends in paint and gasoline lead exposure are also strongly associated with subsequent trends in murder rates going back to 1900. The findings on violent crime and unwed pregnancy are consistent with published data describing the relationship between IQ and social behavior. The findings with respect to violent crime are also consistent with studies indicating that children with higher bone lead tend to display more aggressive and delinquent behavior. This analysis demonstrates that widespread exposure to lead is likely to have profound implications for a wide array of socially undesirable outcomes. PMID- 10845778 TI - Male and female rate differences in carpal tunnel syndrome injuries: personal attributes or job tasks? AB - Carpal tunnel syndrome (CTS) exacts a significant toll on the health and productivity of American workers. In 1996, 29,937 workers lost time from work due to CTS. Half of these were out for more than 25 days, compared to a median of 5 days for all injuries/illnesses. There are striking gender disparities in CTS rates. Overall, three times more women suffer from CTS than men. While some authors have emphasized the role of gender attributes in this risk disparity, the multifactoral causes of CTS, and the sex segregation of women into jobs with high risk tasks, may be obscuring the work-related contributions to CTS risk. We argue here that men and women doing the same work tasks will have similar rates of CTS. To examine this premise, we used Bureau of Labor Statistics injury counts (numerator) and census data from the Current Population Survey (denominator) to determine injury rates of CTS for both men and women in six high-risk occupations: (1) assembler, (2) laborer-nonconstruction, (3) packaging and filling machine operators, (4) janitors and cleaners, (5) butchers and meat cutters, and (6) data entry keyers. Variable job tasks exist within five of the six high-risk occupational titles. Among those five, the male to female (M:F) risk rate ratio ranged from 0.29 to 0.50. However, the sixth occupational title, data entry keyers, which requires a single physical task, had a risk rate ratio of 1.06. This suggests that an equal risk between genders exists when the occupational tasks (exposure) are truly similar. Job task analysis unmasks potential biases that may wrongly attribute disproportionate CTS rates to gender attributes. This focus on gender attributes fails all workers because preventive interventions in the workplace are deferred. It fails women disproportionately because they are overrepresented in jobs at high risk for CTS. PMID- 10845779 TI - Female lung cancer mortality and sex ratios at birth near a petroleum refinery plant. AB - This study was conducted to assess whether female mortality from lung cancer is associated with residence in communities adjacent to a petroleum refinery plant and whether petroleum air pollution could affect the sex ratios of births. The Kaohsiung Refinery of the Chinese Petroleum Corp. is the oldest oil refinery in Taiwan and is located between the Tso-Ying and the Nan-Tzu municipalities. Standardized mortality ratios (SMRs) for female lung cancer and sex ratios of births were calculated for each municipality for the years 1971-1996. Cumulative sum techniques were used to detect the occurrence of changes in the SMRs. The study results show that mortality from female lung cancer rose gradually about 30 to 37 years after the operation of a petroleum refinery plant began. However, the association between exposure to the petroleum air pollution and abnormal sex ratios at birth was not significant. PMID- 10845780 TI - Semi-occupational exposure to lead: a case study of child and adolescent street vendors in Istanbul. AB - This study analyzed lead levels in scalp hair of child and adolescent street vendors in Istanbul. The mean lead in hair concentration found among children exposed to intensive motor traffic, 11.82 +/- 6.89 microg/g, was over five times higher than that in the control group, 2.70 +/- 2.30 microg/g. Vendors from a location with higher traffic density had significantly higher lead in hair levels, than vendors from a location with lower traffic density (14.18+/- 5.33 and 9.86 +/- 7.54 microg/g, respectively). The results pointed to the dose response effect between traffic intensity and lead in hair concentration. Despite low lead content in gasoline, an intensive exposure to motor traffic may be a nonnegligible source of lead and poses a considerable health risk for individuals who work in particularly busy locations. PMID- 10845781 TI - Decrease of PCDD/F levels in human blood--trend analysis for the German population, 1991-1996. AB - More than 500 whole blood samples of normal subjects from Germany collected in 1991-1996 have been analyzed for polychlorinated dibenzo-p-dioxins (PCDD) and dibenzofurans (PCDF) by capillary gas chromatography/high-resolution mass spectrometry. Over the examined time period a continuous decrease of the PCDD/F concentrations in human blood was observed. The mean levels found were about 42.7 pg I-TEq/g (lipid basis) in 1991 and 20.7 pg I-TEq/g (liquid basis) in 1996 [median: 40.8 and 19.2]. A reduction to about half was found for most congeners. Each 1-year subset of the entire collective shows a positive correlation of the PCDD/F blood levels with age for most of the congeners, the sum values, and the calculated toxicity equivalents. For statistical evaluation a multiplicative model was used: Concentration in blood = A x ageB. The correlation is mostly pronounced for lower chlorinated PCDD and for 2,3,4,7,8-PentaCDF. PMID- 10845782 TI - Accumulation of cadmium, zinc, and copper in maternal blood and developmental placental tissue: differences between Finland, Estonia, and St. Petersburg. AB - Cadmium, zinc, and copper from placental tissue and blood samples at the first trimester (n = 64) and at term (n = 152) were analyzed; the welfare of newborns and placental 7-ethoxycoumarin O-deethylase (ECOD) activities in vitro were determined. The study material was collected from Finland, Estonia, and Russia. The results demonstrate that Cd starts to accumulate in the placenta during the first trimester and that Zn and Cu contents were significantly higher at the first trimester than at term. Among nonsmokers a negative correlation was found between placental Cu content and birth weight of neonates. Among smokers a positive correlation between placental Zn content and birth weight and ECOD activity was found. The birth weights correlated inversely with the length of time the mothers smoked. The highest Cd concentrations were detected in the samples collected from St. Petersburg. The data demonstrate an inverse accumulation of Zn and Cd throughout the pregnancy in the placenta and maternal blood samples. Zn may act as a positive marker or even an enzymatic enhancement for the human placental vital functions. Smoking, parity, age, and especially the place of residence affect the Cd, Zn, and Cu contents and ratios in placenta and mother's blood. PMID- 10845783 TI - A model for predicting the frequency of high pesticide exposure events in the Agricultural Health Study. AB - The frequency of self-reported high pesticide exposure events (HPEE) has been recorded in the NCI/EPA/NIEHS Agricultural Health Study (AHS). Fourteen percent (14%) of the enrolled applicators responding reported "an incident or experience while using any pesticide which caused an unusually high exposure." These data show, as expected, that the probability of a report of an HPEE increases with the cumulative number of days of pesticide application reported by the applicator. We have developed a three-parameter model that predicts the risk odds ratio (OR) of an HPEE as a function of the number of days that pesticides are applied. These events are costly in terms of resulting health-care visits, lost time from work, and potential risk for cancer and other chronic diseases. We propose that failure to carefully follow all the pesticide manufacturer's label requirements, inexperience, and random events (i.e., breaking hose) are the three factors responsible for the events observed. Special precautions for new or infrequent users of pesticides are indicated. PMID- 10845784 TI - The effects of chlorothalonil on oyster hemocyte activation: phagocytosis, reduced pyridine nucleotides, and reactive oxygen species production. AB - The production of reactive oxygen species (ROS) by a putative NADPH oxidase-like enzyme system is thought to contribute to antimicrobial activity in oyster hemocytes. NADPH oxidase in vertebrate phagocytes generates superoxide anion from molecular oxygen and NADPH, which is then converted to additional ROS, including H2O2 and HOCl. The fungicide chlorothalonil (TCIN) is a thiol-reactive compound that binds to protein sulfhydryl groups, which can result in enzyme inactivation. NADPH oxidase, containing several sulfhydryl groups, is a potential target of TCIN. Previous studies have demonstrated that in vitro exposure of fish (Morone saxatilus) macrophages to TCIN (10-500 microg/L) suppressed immunostimulated ROS and baseline NAD[P]H concentration but did not inhibit phagocytosis; the production of NADPH in stimulated cells was decreased only at the highest concentration. In this study, we evaluated the effects of TCIN (10-500 microg/L) on oyster hemocyte functions. As with striped bass macrophages, in vitro exposure to TCIN suppressed hemocyte ROS production in a dose-dependent manner, but did not affect phagocytosis. In contrast to the striped bass data, baseline NAD[P]H concentration was relatively unaffected and immunostimulated NAD[P]H production was marginally suppressed at the higher exposure concentrations. Despite these minor differences, these results suggest that TCIN may also be inhibiting an NAD[P]H oxidase-like enzyme in oyster hemocytes. PMID- 10845785 TI - Adsorption equilibria and overloaded band profiles of basic drugs in a reversed phase system. AB - Single-solute adsorption equilibrium isotherms of three basic drugs: buspirone, doxepin and diltiazem were determined by frontal analysis in a reversed-phase system composed of an octadecylsilica packing material and a buffered mobile phase containing acetonitrile. The adsorption data were fitted to the bi-Langmuir model. Within the framework of this model, the adsorption of the drugs is assumed to occur on two distinct kinds of sites with different average adsorption energies. The data are consistent with the assumption that the low energy sites account for the hydrophobic interactions between the solutes and the chemically bonded alkyl chains and the high energy sites account for the ion-exchange interactions between the residual active silanols and the protonated bases. Multisolute, overloaded band profiles were also measured for the three binaries and for mixtures of the three drugs. Theoretical band profiles were calculated using the equilibrium dispersive model and the ideal adsorbed solution theory model which uses the parameters determined from the correlation of the single solute adsorption data. Good agreement was found between the experimental and calculated overloaded band profiles. PMID- 10845786 TI - Protein retention in ion-exchange chromatography: effect of net charge and charge distribution. AB - The charge regulated slab model is used to evaluate the salt dependence of the retention of Staphylococcal nuclease A and its mutants in cation-exchange chromatography. An important feature of this work is that the net charge of the proteins is varied in two different ways: (a) by changing the eluent pH so that the charges are created by protonation and (b) by point mutation at position 116. Since the structure of Staphylococcal nuclease and the mutants are known, the pH dependence of retention data of the different mutants gives detailed insights into the retention mechanism. Experimental results show that the salt dependence of retention is affected more strongly by changes of the eluent pH than by point mutations. This implies that the amino acid in position 116 has only a moderately strong interaction with the stationary phase surface and that a patch on one side of the protein surface is mainly responsible for the electrostatic interaction with the surface. PMID- 10845787 TI - Characterization of C18-bonded liquid chromatographic stationary phases by Raman spectroscopy: the effect of mobile phase composition. AB - Raman spectroscopy is used to examine the effect of mobile phase composition on the orientation of octadecyl-bonded silica-based reversed-phase liquid chromatographic (RPLC) stationary phase ligands. The effect of ligand bonding density is also investigated. The present experimental set-up utilizes a direct, noninvasive, on-column approach to examine the solvent dependent conformational behavior of the bonded ligands under flow-rate and back pressure conditions similar to those used during conventional RPLC measurements. Neat, single component, mobile phase solvents including water, acetonitrile, methanol and chloroform are used to investigate the hypothesized collapsing and extension of stationary phase ligands with changes in mobile phase composition. No evidence of phase collapse was observed upon changing the mobile phase composition from an organic to an aqueous content. Also, Raman spectroscopic measurements allowed the differentiation between associated and free acetonitrile solvent. PMID- 10845788 TI - Characterization of C18-bonded liquid chromatographic stationary phases by Raman spectroscopy: the effect of temperature. AB - This study represents the first time that both the mobile phase composition and the temperature are simultaneously controlled to examine silica-bonded octadecylsilyl (C18) ligands spectroscopically at typical liquid chromatographic (LC) mobile phase flow-rates and back-pressures. Raman spectroscopy is used to characterize the behavior of the C18 bonded ligands equilibrated at temperatures from 45 to 2 degrees C in neat, single-component, mobile phase solvents including: water, acetonitrile, methanol, and chloroform. In addition, the effect of stationary phase ligand bonding density is examined by using two different monomeric reversed-phase liquid chromatographic (RPLC) stationary phases, a 2.34 and a 3.52 micromol m(-2) Microporasil C18 stationary phase, under identical conditions. The direct, on-column, spectroscopic analysis used in this study allows direct evaluation of the temperature-dependent behavior of the bonded C18 ligands. The temperature-dependent ordering of the stationary phase ligands is examined to determine if the ligands undergo a phase transition from a less ordered "liquid-like" state at higher temperatures to a more-ordered "solid-like" state at lower temperatures. A discrete phase transition was not observed, but rather a continual ordering as temperature was lowered. PMID- 10845789 TI - Separation of aromatic isomers on cyclophane-bonded stationary phases. AB - A cyclophane (CP66)-bonded silica gel stationary phase (CP66-SP) was prepared and the retention of water-insoluble hydrophobic compounds on it was investigated in comparison with that on the CP44-bonded stationary phase (CP44-SP) reported previously. Like CP44-SP, it retained aromatic compounds more strongly than the corresponding alicyclic compounds, as was expected by the cavity size of the cyclophane. The CP66-SP also showed isomer-selectivity for monosubstituted and disubstituted naphthalenes, but its selectivity was perfectly reversed to that of the CP44-SP. On the CP66-SP, isomers having methyl and ethyl groups at beta position were eluted prior to those having groups at alpha-position, whereas on the CP44-SP beta-substituted naphthalenes were retained more strongly than alpha substituted ones. Isomers of three- and four-ring aromatic compounds were also separated on these cyclophane-bonded stationary phases. The retention order on the CP66-SP was almost opposite to that on the CP44-SP; on the CP66-SP, the retention order was phenanthrene > anthracene, and chrysene > 1,2-benzanthracene > 2,3-benzanthracene, whereas on the CP44-SP, anthracene > phenanthrene, and 2,3 benzanthracene > chrysene > 1,2-benzanthracene. The retention mechanism of aromatic compounds is discussed on the basis of the structure of the cyclophane involved complex. PMID- 10845790 TI - Retention versatility of silica-supported comb-shaped crystalline and non crystalline phases in high-performance liquid chromatography. AB - Silica-supported comb-shaped polymer (Sil-ODA18) provided a specific selectivity towards aromatic and non-aromatic solutes. Selectivity of Sil-ODA18 and ODS (polymeric and monomeric) columns towards polycyclic aromatic hydrocarbons and geometric isomers have been discussed in respect of molecular shapes and sizes. The retention versatility of this phase arises from the orientation change on transition from the ordered crystalline to the less ordered non-crystalline state. Selectivity in the crystalline and non-crystalline regions of Sil-ODA18 is similar to that of polymeric and monomeric ODS phases, respectively. Sil-ODA18 column also exhibited the characteristic temperature dependency of liquid crystalline phases, i.e., an increase in retention with increasing temperature on transition from crystalline to isotropic state. PMID- 10845791 TI - Alkyltrimethylammonium surfactant-mediated extractions: characterization of surfactant-rich and aqueous layers, and extraction performance. AB - Most surfactants employed for extraction purposes contain strongly absorbing chromophores, and therefore cannot be used with the ultraviolet-visible HPLC detector because of the high background created. Alkyltrimethylammonium surfactants, which do not have strongly absorbing chromophores, have shown promise as an extractant compatible with HPLC-ultraviolet-visible detection. In our extraction procedure, alkyltrimethylammonium surfactants are added to a sample containing organic analytes in distilled water. Sodium chloride is next added, then the entire sample is shaken. Before centrifugation, 1-octanol is added to aid in the two phase formation of surfactant-rich and aqueous phases. In this paper, we present the results of our studies on the extraction behavior of an alkyltrimethylammonium surfactant technique using various organic compounds as test probes. Specifically studied are the extraction behavior of organic bases, isomers of varying polarity and a zwitterionic species that has different charges at various pH values. Results from multiple extractions to obtain quantitative recovery of analytes is also presented. The composition of each phase is elucidated through the interpretation of data obtained from thermogravimetric and carbon, hydrogen and nitrogen (CHN) instrumentation. PMID- 10845792 TI - Study of the adsorption behavior of the enantiomers of 1-phenyl-1-propanol on a cellulose-based chiral stationary phase. AB - Using single-step frontal analysis, we measured single-component and competitive adsorption isotherm data for the two enantiomers of 1-phenyl-1-propanol (PP). These experimental data were fitted to several competitive bi-Langmuir models (with 8, 6, 5 and 4 parameters) and to the competitive Langmuir model. The latter model accounted well for the behavior of both PP enantiomers on Chiracel OB (cellulose tribenzoate coated on silica gel). The parameters obtained were used in numerical calculations to predict the band profiles of the two single components and of their mixtures under overloaded conditions. The equilibrium dispersive model provides satisfactory results, with minor differences between the calculated and the experimental profiles. These differences became negligible when a more complex kinetic model was used, with a concentration-dependent rate coefficient. PMID- 10845793 TI - Adsorption behavior and prediction of the band profiles of the enantiomers of 3 chloro-1-phenyl-1-propanol. Influence of the mass transfer kinetics. AB - The single-component and competitive adsorption isotherms of the enantiomers of 3 chloro-1-phenyl-1-propanol were measured by frontal analysis. The stationary phase was a cellulose tribenzoate coated on silica, the mobile phase an n-hexane ethyl acetate (95:5) solution. The adsorption data measured fitted well to the Langmuir isotherm model. The band profiles of single components and of their mixtures were calculated using the equilibrium-dispersive model. These profiles were found to match quite satisfactorily the experimental band profiles. However, the agreement between calculated and experimental band profiles was significantly improved when a more complex model taking into account the mass transfer kinetics was used. The mass transfer rate coefficients, k(f), for both single components were determined by using the transport-dispersive model of chromatography. The coefficients obtained were used to predict the band profiles of mixtures of the two enantiomers to good agreement. PMID- 10845794 TI - Determination of trace cadmium in environmental water samples using ion interaction reversed-phase liquid chromatography with fluorescence detection. AB - An ion-interaction reversed-phase liquid chromatographic method has been developed for the determination of cadmium at low microg/l concentrations in environmental water samples. Cadmium and other matrix metals were separated through on-column complexation with 8-hydroxyquinoline sulphonate, using an octadecylsilica column and a mobile phase containing 15% acetonitrile, 10-13 mM tetrabutylammonium hydroxide, 5 mM 8-hydroxyquinoline 5-sulphonic acid and 10 mM acetic acid-acetate buffer (pH 4.8-5.4). Under the above conditions Cd(II) could be easily resolved from excess concentrations of matrix metals and could be detected at concentrations as low as 2 microg/l using fluorescence detection at 500 nm (based upon a 100-microl injection). The method showed a slightly curved detector response over the range of interest [up to 1 mg/l Cd(II)] and was successfully applied to the determination of trace Cd(II) in water samples containing large excesses of Mg(II) and Zn(II) and other matrix metals. PMID- 10845795 TI - Application of electron-attachment reactions to enhance selectivity of electron capture detector for nitroaromatic explosives. AB - The differences in the extent of electron-attachment reactions between thermal electrons and selected classes of organic molecules with high electron affinities were investigated. The investigations showed that interactions of thermal electrons with nitroaromatic compounds lead to the formation of neutral products with very low electron affinities. By contrast, a number of other analytes with high electron affinities such as polyhalogenated organic compounds, lead to products with high electron affinities. This difference was exploited to differentiate between nitroaromatic and polychlorinated organic compounds with a tandem arrangement consisting of two electron-capture detectors connected in series with an electron-attachment reactor. PMID- 10845796 TI - Dynamic microwave-assisted extraction. AB - An apparatus for extraction of solid matrices has been constructed which utilizes a microwave technique for heating in a dynamic mode. During the extraction, fresh solvent is continuously pumped through the extraction cell, which is maintained at a slight overpressure in order to keep the solvent in a liquid state. The extraction efficiency, which can be easily monitored, has been investigated in a factorial design and validated for polycyclic aromatic hydrocarbons in a reference sediment sample (EC-1). Important parameters were found to be temperature and duration of extraction. Flow-rate had no significant first-order effect on the recovery, but interaction effects with flow-rate were found to be significant. The dynamic microwave-assisted extraction apparatus was demonstrated to yield recoveries equivalent to Soxhlet extraction, but in a much shorter time. Each extraction of EC-1 typically takes 40 min. PMID- 10845797 TI - Determination of chlorophenoxy acid herbicides in water by in situ esterification followed by in-vial liquid-liquid extraction combined with large-volume on-column injection and gas chromatography-mass spectrometry. AB - A new approach for rapidly analysing chlorophenoxy acid herbicides in water is presented. The chlorinated acids are derivatised with dimethyl sulphate in the water sample itself (800 microl) and, next, the methyl esters are extracted with 800 microl of n-hexane. A 200-microl volume of the extract is injected into the GC-MS system. The miniaturisation of both the methylation and extraction steps could be implemented because of the use of large-volume on-column injection and mass spectrometric detection. The optimisation of the methylation reaction for the simultaneous determination of (3,6-dichloro-2-methoxy)benzoic acid, (2-methyl 4-chlorophenoxy)- and (2,4-dichlorophenoxy)acetic acids, (+/-)-2-(4-chloro-2 methylphenoxy)- and 2-(2,4-dichlorophenoxy)propanoic acids and 4-(4-chloro-2 methylphenoxy)- and 4-(2,4-dichlorophenoxy)butyric acids showed that tetrabutylammonium salts act as catalysts. Addition of sodium hydroxide was required to obtain quantitative reaction yields for 4-(4-chloro-2-methylphenoxy)- and 4-(2,4-dichlorophenoxy)butyric acids. The methylation-cum-extraction procedure takes only 3 min per sample for a batch of seven samples. Linear calibration plots were obtained for the complete procedure and the limits of detection were of 10-60 ng/l with a signal-to-noise ratio (S/N) of 6. Relative standard deviations ranged from 8 to 15% (n=7) for analyte concentrations of 0.5 microg/l in surface water. PMID- 10845798 TI - Utilization of fat retainers in supercritical fluid extraction for the selective extraction of polychlorinated biphenyls from a model fat sample. AB - Two common fat retainers used in supercritical fluid extraction--basic alumina and the silica based adsorbent Florisil--were investigated using lard fat as model material. With a fat retainer in the extraction cell it was possible to obtain fat-free time windows. Activation by heating did not influence the length of the time windows, while deactivation of the retainers with 10% water (w/w) drastically decreased the fat retaining capabilities. The influence of modifier addition was also investigated. Finally, a method was developed, where basic alumina was utilized to selectively extract polychlorinated biphenyls (PCBs) from a model fat sample, containing PCBs, triglycerides and phospholipids. The PCBs could be quantitatively extracted in a totally fat-free time window. PMID- 10845799 TI - Modeling the velocity field of the electroosmotic flow in charged capillaries and in capillary columns packed with charged particles: interstitial and intraparticle velocities in capillary electrochromatography systems. AB - Mass transfer systems based on electrokinetic phenomena (i.e., capillary electrochromatography (CEC)) have shown practical potential in becoming powerful separation methods for the biotechnology and pharmaceutical industries. A mathematical model has been constructed and solved to describe quantitatively the profiles of the electrostatic potential, pressure, and velocity of the electroosmotic flow (EOF) in charged cylindrical capillaries and in capillary columns packed with charged particles. The results obtained from model simulations (i) provide significant physical insight and understanding with regard to the velocity profile of the EOF in capillary columns packed with charged porous particles which represent systems employed in CEC, (ii) provide the physical explanation for the experimental results which indicate that the velocity of the EOF in capillary columns packed with charged porous particles is a very weak function (it is almost independent) of the diameter of the particles, and (iii) indicate that the intraparticle velocity, nu(p,i), of the EOF can be greater than zero. The intraparticle Peclet number, Pe(int rap), for lysozyme was found to be greater than unity and this intraparticle convective mass transfer mechanism could contribute significantly, if the appropriate chemistry is employed in the mobile liquid phase and in the charged porous particles, in (a) decreasing the intraparticle mass transfer resistance, (b) decreasing the dispersive mass transfer effects, and (c) increasing the intraparticle mass transfer rates so that high column efficiency and resolution can be obtained. Furthermore, the results from model simulations indicate that for a given operationally permissible value of the applied electric potential difference per unit length, Ex, high values for the average velocity of the EOF can be obtained if (1) the zeta potential, zeta(p), at the surface of the particles packed in the column has a large negative magnitude, (2) the value of the viscosity, mu, of the mobile liquid phase is low, (3) the magnitude of the dielectric constant, epsilon, of the mobile liquid phase is reasonably large, and (4) the combination of the values of the concentration, C(infinity), of the electrolyte and of the dielectric constant, epsilon, provide a thin double layer. The theoretical results for the velocity of the EOF obtained from the solution of the model presented in this work were compared with the experimental values of the velocity of the EOF obtained from a fused-silica column packed with charged porous silica C8 particles. Systems with four different particle diameters and three different concentrations of the electrolyte were considered, and the magnitude of the electric field was varied widely. The agreement between theory and experiment was found to be good. PMID- 10845800 TI - Chiral separation of homocysteine by derivatization with 4-aminosulfonyl-7-fluoro 2,1,3-benzoxadiazole followed by capillary electrophoresis using gamma cyclodextrin. AB - Homocysteine was derivatized with 4-aminosulfonyl-7-fluoro-2,1,3-benzoxadiazole (ABD-F) to form an inclusion complex with cyclodextrin and to facilitate UV detection. ABD-homocysteine showed interaction with beta- and gamma-cyclodextrin in capillary electrophoresis at pH 2.25 as indicated by the decreased migration time. However, chiral separation of D,L-ABD-homocysteine was observed using gamma CD only. Optimal separation was obtained at pH 2.25, 50 mM gamma-CD concentration, and 20 kV applied voltage. L-ABD-Homocysteine migrated faster than the D-isomer as demonstrated by a spiking experiment using dithiothreitol-reduced L-homocystine. PMID- 10845801 TI - Influence of ring opening-closure equilibrium of oxanine, a novel damaged nucleobase, on migration behavior in capillary electrophoresis. AB - Oxanine (Oxa) is a novel damaged nucleobase which is generated from guanine by HNO2 or NO. As a fundamental study for detection of Oxa formed in vivo, capillary zone electrophoresis (CZE) and micellar electrokinetic chromatography (MEKC) have been tested by changing the pH of the running buffer. At pH 7, CZE did not separate Oxa from seven other nucleobases, and MEKC separated Oxa but their peaks migrated close together. In both the techniques, an extreme peak broadening occurred for Oxa around pH 9 and a good peak separation was achieved at pH 12. The behavior of the Oxa peak is discussed in relation to the unique multistep acid-base equilibria of Oxa. PMID- 10845802 TI - Analysis of baclofen by capillary electrophoresis with laser-induced fluorescence detection. AB - A new analytical method for baclofen (4-amino-3-p-chlorophenylbutyric acid) based on capillary electrophoretic separation and laser-induced fluorescence detection has been developed. Naphthalene-2,3-dicarboxaldehyde was used for precolumn derivatization of the non-fluorescent drug. Optimal separation and detection were obtained with an electrophoretic buffer of 50 mM sodium borate (pH 9.5) and a He Cd laser (excitation at 442 nm, emission at 500 nm). Linearity (r > or = 0.99) over three orders of magnitude was generally obtained and the concentration limit of detection was in the nanomolar level. Coupled with a simple cleanup procedure, the method was successfully applied to the analysis of baclofen in human plasma. Recovery of spiked baclofen in plasma was 98%. The relative standard deviation values on peak size and migration time were 7.9% and 0.4%, respectively. The limit of detection of baclofen in plasma was 10 ng/ml. PMID- 10845803 TI - Is ICI 182,780 an antiprogestin in addition to being an antiestrogen? AB - The pure antiestrogen ICI 182,780 has been shown to have antiprogestin activity in reporter gene constructs. Cell lines, naturally devoid of progesterone receptors (PR) were transfected with either the A or B forms of the human PR and a luciferase construct driven by a progesterone-response element (PRE). Because this system is an artificial one, our purpose was to determine whether these observations could be made in a human breast cancer cell line, naturally containing PR. We further evaluated the dose-response of ICI 182,780 and RU-486 (mifepristone) on PR and estrogen receptors (ER) in the presence of either progesterone, norgestrel or estradiol. These effects were measured using immunoassays for prostate-specific antigen (PSA) and human glandular kallikrein (hK2) and pS2. We found that ICI 182,780 blocked progesterone-stimulated PSA and hK2 production 100% at 10(-5) M, which decreased significantly by 10-6 M. This inhibition did not occur when norgestrel was the progestin used. RU-486 showed 100% blockade for both progestins at all concentrations used. We concluded that the antiprogestin activity of ICI 182,780 exists for progesterone only. This weak antiprogestin activity may be unlikely to have significant clinical implications. PMID- 10845804 TI - Framing of outcome and probability of recurrence: breast cancer patients' choice of adjuvant chemotherapy (ACT) in hypothetical patient scenarios. AB - PURPOSE: To examine the effects of framing of outcome and probabilities of cancer occurrence on the treatment preference which breast cancer patients indicate for hypothetical patient scenarios. METHODS: A modified version of the Decision Board Instrument (Levine et al. 1992) was administered to 35 breast cancer patients with past ACT experience. Patients expressed their choice regarding ACT for six scenarios which were characterized by either negative or positive framing of outcome and by one of the three levels of probability of recurrence (high, medium, low). RESULTS: The framing had no influence on ACT choices over all three probability levels. The majority chose ACT for high and medium risk and one third switched from ACT to No ACT in the low-risk condition. This switch was statistically significant. CONCLUSION: Hypothetical treatment decisions against ACT occur only when the probability of recurrence is low and the benefit of ACT is small. This finding for patients with past experience of ACT is similar to those reported for other oncological patient groups still in treatment. PMID- 10845805 TI - Progression in MCF-7 breast cancer cell tumorigenicity: compared effect of FGF-3 and FGF-4. AB - The transforming properties of fibroblast growth factor 3 (FGF-3) were investigated in MCF7 breast cancer cells and compared to those of FGF-4, a known oncogenic product. The short form of fgf-3 and the fgf-4 sequences were each introduced with retroviral vectors and the proteins were only detected in the cytoplasm of the infected cells, as expected. In vitro, cells producing FGF-3 (MCF7.fgf-3) and FGF-4 (MCF7.fgf-4) displayed an amount of estrogen receptors decreased to around 45% of the control value. However, MCF7.fgf-3 cell proliferation remained responsive to estradiol supply. The sensitivity of the MCF7.fgf-4 cells, if existant, was masked by the important mitogenic action exerted by FGF-4. In vivo, the MCF7.fgf-3 and MCF7.fgf-4 cells gave rise to tumors under conditions in which the control cells were not tumorigenic. Supplementing the mice with estrogen had the paradoxical effect of totally suppressing the start of the FGF-3 as well as the FGF-4 tumors. Tumorigenicity in the presence of matrigel was similar for MCF7.fgf-3 and control cells and was increased by estrogen supplementation. Once started, the MCF7.fgf-4 tumors grew with a characteristic high rate. Remarkably, FGF-4 but not FGF-3, stimulated the secretion of vascular endothelial growth factor (VEGF165) without altering the steady-state level of its mRNA, suggesting a possible regulation of VEGF synthesis at the translational level in MCF7 cells. The increased VEGF secretion is probably involved in the more aggressive phenotype of the MCF7.fgf-4 cells while a decreased dependence upon micro-environmental factors might be part of the increased tumorigenic potential of the MCF7.fgf-3 cells. PMID- 10845806 TI - Resection margin status in lumpectomy specimens of infiltrating lobular carcinoma. AB - AIMS: To study the status of resection margins in specimens from patients with infiltrating lobular carcinoma (ILC) treated with lumpectomy. MATERIALS AND METHODS: Sixty-six consecutive cases of ILC were compared with the same number of consecutive cases of infiltrating ductal carcinoma (IDC). All cases were treated with lumpectomy. RESULTS: ILCs were divided into 42 cases of typical ILC, 15 variants of ILC (alveolar or solid types) and 9 cases of mixed ILC and IDC. These groups were associated with positive or close resection margins in 22 (52%), 5 (33%) and 3 (33%) cases, respectively. For the group of IDC with partial mastectomies, matched for patient's age and tumor size, positive or close resection margins were observed in 26%. ILCs, measuring less than 2 cm in greatest diameter and having low nuclear grade, had rates of positive or close margins comparable with those of IDC. Typical ILCs, measuring more than 2 cm in diameter, had rates of positive or close margins of 70%. All cases with a positive extensive intraductal component had positive margins. Furthermore, in all types of ILC, tumors with a high nuclear grade tended to be associated with a high rate of positive margins. CONCLUSIONS: The status of resection margins in lumpectomy specimens for infiltrating lobular carcinoma is related to the extensive intraductal component status, tumor size and grade, and the presence of variants of ILC or mixed ILC and IDC. Most of these factors can be determined preoperatively by mammography and histopathological evaluation of breast core biopsies, therefore, aiding in planning the surgical strategy of mastectomy. PMID- 10845808 TI - Correlation of three-dimensional magnetic resonance imaging with precise histopathological map concerning carcinoma extension in the breast. AB - This study was initiated to clarify the ability of magnetic resonance imaging (MRI) in defining breast carcinoma extension by comparing MRI to detailed histopathological analysis. Mastectomy (n = 14) or quadrantectomy (n = 44) specimens were sub-serially sectioned and mapped in detail in 58 breast cancer patients. Morphologically, we classified the lesions utilizing MRI into three patterns in relation to their histology. Numerically, we assessed the maximum distance of carcinoma extension using MRI, mammography, and ultrasonography (US). Linear regression was calculated for each of the three imaging measurements versus histopathological measurements. Three imaging patterns were observed by MRI, (1) localized (n = 30), (2) segmentally extended (n = 19), and (3) irregularly extended (n = 5). The localized pattern showed a distinct focal mass, but in 10 cases, microscopic ductal carcinoma in situ (DCIS), or invasive lobular carcinoma, which were not depicted by MRI, existed. The segmentally extended pattern showed diffuse enhancement along duct-lobular segments, forming a 'cone' shape. Histologically, pure (n = 4) or predominant (n = 10) DCIS was distributed segmentally. The irregularly extended pattern showed thick branches extending out from the index tumor which were histologically revealed to be stromal invasion of ductal carcinoma. From the results of linear regressions, MRI was the most accurate modality in histologically measuring the extent of the cancer. When cases were limited to patients who were classified into segmentally or irregularly extended pattern by MRI (n = 24), MRI was more accurate than mammography and US, even if they were combined (P < 0.05). MRI may provide additional information concerning carcinoma extension prior to surgery, especially in patients classified into 'extended patterns' by MRI. PMID- 10845807 TI - Detection of microsatellite alterations in nipple discharge accompanied by breast cancer. AB - Nipple discharge in breast cancer cases was examined loss of heterozygosity (LOH). DNA samples were extracted from both supernatant and cell pellet components of the discharge, and examined for LOH at microsatellite markers, D11S1818, D11S2000, D16S402, D16S504, D16S518, D17S520, and D17S786. At least one LOH was found in either the supernatant or cell pellet in seven out of 10 patients (70%). Five of seven samples, which were cytologically negative, were LOH positive, and only one case, which was cytologically positive, showed no LOH on the markers examined. All three samples, which were judged 'negative' by CEA measurement (<400 ng/ml), were LOH positive. This method could be a useful novel diagnostic modality for nonpalpable breast cancer with nipple discharge. PMID- 10845809 TI - Randomized phase II study of FEC day 1 + 8 and FEC day 1 in patients with advanced breast cancer. AB - BACKGROUND: Dose-intensive chemotherapy regimens without stem cell support have not resulted in an improved survival compared to standard dose regimens in patients with metastatic breast cancer. Combinations of an anthracycline, cyclophosphamide and 5 fluorouracil are still standard in such patients. The aim of this study was to investigate the two different schedules of epirubicin in a standard dose FEC regimen with respect to response and toxicity. MATERIALS AND METHODS: Patients were randomly assigned to receive a day 1 + 8 schedule (5FU and CTX 500mg/m2 day 1, epirubicin 40 mg/m2 day 1 and 8) or a day 1 schedule (5FU, CTX 500 mg/m2 and epirubicin 80 mg/m2 day 1), q day 21, both given without hematopoietic growth factors. A total of 104 eligible patients were analyzed, 52 in each arm. RESULTS AND CONCLUSIONS: A significantly higher relative dose intensity was found for the day 1 schedule compared to the day 1 + 8 schedule. Although the trial was not set up to reliably detect a difference in response rate, this difference in relative dose-intensity in favour of the day 1 schedule does not suggest any improvement in response rate or duration of response for the day 1 schedule. Myelosuppression was severe in the day 1 + 8 schedule. We conclude that a day 1 + 8 FEC schedule has no advantage over a day 1 FEC schedule without hematopoietic growth factors in patients with metastatic breast cancer. PMID- 10845810 TI - Genotoxic effects of the novel mixed antiestrogen FC-1271a in comparison to tamoxifen and toremifene. AB - Tamoxifen has been used for the treatment of breast cancer since the 1970s, but is considered a carcinogen because it has been linked to liver cancer in rats and an increased risk of endometrial cancer in patients. In rats, DNA adducts appear to be responsible for carcinogenesis, but their contribution to carcinogenesis in humans is not clear. FC-1271a and toremifene are mixed antiestrogens similar to tamoxifen. In order to compare the genotoxicity of these different triphenylethylenes, we treated mice for 28 days with 50 mg/kg of either tamoxifen, toremifene, FC- 1271 a or vehicle control. DNA from liver and uterus was assayed by standard 32P-postlabeling and thin layer chromatography for the presence of DNA adducts. Two methods of drug administration (oral and subcutaneous) and two strains of mice were compared and the plasma and tissue concentrations of the drugs and three metabolites of tamoxifen and toremifene were determined. Regardless of the conditions, only tamoxifen-treated mice showed DNA adducts in the liver. Adduct levels did not correlate with drug or metabolite levels and adducts were present even when drug was not detectable. Mice were also treated orally with either 50, 100, or 200 mg/kg of drug for 7 days. Again, adducts were found only in liver tissue of mice treated with tamoxifen, and adduct levels were dose-dependent. In conclusion, the chlorinated triphenylethylene FC-1271a did not cause DNA adducts under various conditions in mice, suggesting a low carcinogenic potential. PMID- 10845811 TI - Antagonists of growth hormone-releasing hormone arrest the growth of MDA-MB-468 estrogen-independent human breast cancers in nude mice. AB - Since antagonists of growth hormone-releasing hormone (GH-RH) inhibit proliferation of various tumors, in this study we investigated the effects of GH RH antagonists MZ-5-156 or JV-1-36 on growth of estrogen-independent MDA-MB-468 human breast cancers xenografted into nude mice. Both GH-RH antagonists administered at a dose of 20 microg/day induced regression of some and growth arrest of other tumors, while control tumors continued to grow. After 5 weeks of therapy with MZ-5-156 or JV-1-36, final volume and weight of MDA-MB-468 tumors were significantly decreased (all p values < 0.001) and serum IGF-I levels as well as tumor IGF-I mRNA expression were reduced as compared with controls. High affinity binding sites for IGF-I were detected by the ligand binding method. Gene expression of human IGF-I receptors, as measured by the RT-PCR, was not significantly different in control and treated MDA-MB-468 tumors. In cell culture, IGF-I did not stimulate, GH-RH slightly stimulated, while MZ-5-156 and JV-1-36 inhibited proliferation of MDA-MB-468 cells known to possess defective insulin and IGF-I receptor signaling. The expression of mRNA for human GH-RH was found in five of 8 tumors treated with GH-RH antagonists, and in one of the five control tumors. These results suggest that GH-RH antagonists inhibit MDA-MB-468 breast cancers possibly through mechanisms involving interference with locally produced GH-RH. PMID- 10845812 TI - Differences in immunoreactivity of estrogen receptor (ER) in tamoxifen-sensitive and -resistant breast carcinomas: preclinical and first clinical investigations. AB - Inherited or acquired tamoxifen resistance is a major constraint in the endocrinological treatment of breast carcinomas. We developed an enzyme immunoassay that discriminates between tamoxifen-sensitive and -resistant tumors. The procedure was established and standardized using two xenografted breast carcinomas--3366 (highly sensitive to tamoxifen) and 3366/TAM (acquired tamoxifen resistance). The latter model was developed by treatment of 3366 tumor-bearing nude mice during serial passaging over 3 years with tamoxifen. Both lines were estrogen receptor (ER) positive (101 or 82 fmol/mg protein), and revealed no differences in the nucleotide sequences of the hormone binding domain of the ER protein. However, while in the sensitive tumors an upregulation of ER levels was registered after estradiol treatment of tumor bearing nude mice, the ER expression in the resistant line remained unchanged. The tamoxifen sensitive and resistant breast carcinoma 3366 differed, additionally, in their immunoreactivity of ER to mAB H222. While an incubation with estradiol or tamoxifen of immobilized ER prepared from cytosols of the sensitive tumors 3366 led to a significant increase in immunoreactivity, samples of resistant tumors failed in the exposition of additional immunologically reactive epitopes. These results were the basis for the development of an assay for determination of the tamoxifen response in patients. Our retrospective results with 38 breast tumors from a tumor bank indicated that patients with an increase of immunoreactivity of ER more rarely had a recurrence while under going tamoxifen therapy compared with patients expecting no increase. However, the data indicate interesting changes occurring with the ER of tam-resistant tumors that are to be explained by further mutational or protein-chemical analysis. PMID- 10845813 TI - Sonographic evaluation of renal failure. AB - Sonography is a critical component of the evaluation of both acute and chronic renal failure; however, most nephrologists have a limited knowledge of this procedure. The acoustic properties, limited spectrum of pathological changes, and ease of visualization of the kidneys, coupled with the safety, simplicity, and low cost of sonography, make it the modality of choice for renal imaging. This review discusses the basics of sonography as they apply to the kidney and describes the findings encountered in the more common causes of renal failure. Although many sonographic findings are nonspecific, their diagnostic use is greatly enhanced by a familiarity with the clinical presentation and a thorough understanding of renal pathophysiological characteristics. Therefore, nephrologists should be knowledgeable about renal sonography and participate in its interpretation. PMID- 10845814 TI - High prevalence of alcoholism in dialysis patients. AB - Alcoholism is one of the most common psychosocial disorders, affecting approximately 10% of the general population. The impact of alcoholism on the care of patients with other medical illnesses has not been addressed in many of these populations, including patients with end-stage renal disease (ESRD) undergoing hemodialysis. We set out to determine the prevalence of alcoholism in an urban hemodialysis population and ascertain whether alcoholism had an effect on compliance in this population. One hundred sixty-three urban hemodialysis patients were screened using the Michigan Alcoholism Screening Test (MAST), a 25 item questionnaire that has been validated in multiple trials. Forty-five patients (27.6%) scored 5 or greater on the MAST. The MAST-positive subjects were younger (age, 55 +/- 15 years versus 64 +/- 13 years) and tended to be men (58% versus 43%). There was no significant difference in the incidence of diabetic kidney disease; however, there were significantly more human immunodeficiency virus (HIV)-positive patients in the MAST-positive group. The dietary compliance measures of predialysis potassium or phosphorus levels did not differ between the two groups. A trend toward lower serum albumin level was evident in the men in the MAST-positive group (3.75 +/- 0.57 versus 3.91 +/- 0.30 g/dL; P = 0.0212). In conclusion, there is a high prevalence of alcoholism in the urban dialysis population. Alcoholic patients with ESRD are younger and tend to be men. HIV positive patients with ESRD have a high prevalence of concomitant alcoholism. Compliance indicators of predialysis potassium and phosphorus levels are not affected. However, nutritional status, measured by serum albumin level, tends to be poorer in the alcoholic group. PMID- 10845815 TI - Survival after acute myocardial infarction in patients with end-stage renal disease: results from the cooperative cardiovascular project. AB - Cardiovascular disease (CVD) is the most common cause of death in patients with end-stage renal disease (ESRD). The optimal management strategy in this population is unknown. We studied 640 patients with ESRD and acute myocardial infarction during 1994 to 1995 as part of the Health Care Financing Administration's Cooperative Cardiovascular Project. The majority of patients were treated with medical therapy alone, 46 patients (7%) were treated with percutaneous transluminal coronary angioplasty (PTCA), and 29 patients (5%) underwent coronary artery bypass grafting (CABG). Patient characteristics and comorbid conditions were similar among the three groups. The overall 1-year mortality rate was 53%. Advanced age, low or high body mass index, history of peripheral vascular disease or stroke, the inability to walk independently, and several indicators of cardiac dysfunction were associated with an increased relative risk (RR) for death. Survival curves differed significantly by treatment modality, with 1-year survival rates of 45%, 54%, and 69% in the medical therapy alone, PTCA, and CABG groups, respectively (P = 0.03). After adjustment for confounding variables, the RR for death was less (but not significantly so) in the CABG group (RR, 0.6; 95% confidence interval, 0.3 to 1.1). There are no randomized clinical trial data to guide therapy of CVD in patients with ESRD. On the basis of these and other available data, CABG may be the optimal therapy for CVD in ESRD. In light of the exceptionally poor outcomes observed for patients treated with medical therapy alone, it may be premature to dismiss PTCA as a therapeutic option in this population. PMID- 10845816 TI - Potential novel predictors of mortality in end-stage renal disease patients with sleep disorders. AB - Patients with end-stage renal disease (ESRD) have an annual mortality rate exceeding 20%, although some survive many years. The ESRD population has a high incidence of sleep disorders, including sleep apnea and periodic limb movements in sleep (PLMS). Sleep disorders result in sleep deprivation, which can negatively affect immune function and cardiovascular-related outcomes, common causes of death in patients with ESRD. This study examined predictors of mortality in patients with ESRD with sleep problems. Twenty-nine consecutive patients with ESRD reporting disrupted sleep or daytime sleepiness were studied by all-night polysomnography. All patients were followed up until death, transplantation, or study termination. Among the variables studied, including such previously reported predictors as serum albumin level, urea reduction ratio, and hematocrit, only the PLMS index (PLMSI), arousing PLMSI (APLMSI), and total number of arousals per hour of sleep significantly predicted mortality. The 20 month survival rate with a PLMSI less than 20 was greater than 90% versus 50% for a PLMSI of 20 or greater (exact log-rank, P = 0.007). For the deceased versus survivor groups, mean PLMSI was 119.1 versus 19.8 (P = 0.01) and APLMSI was 48.1 versus 7.8 (P = 0.00006), with a mean survival of 10.3 versus greater than 25.5 months, respectively (P = 0.001). Median survival of patients with a PLMSI greater than 80 was only 6 months. PLMSI, APLMSI, and total arousals per hour of sleep were strongly associated with mortality in patients with ESRD with sleep disorders independent of other factors and may be novel predictors of near-term mortality. PMID- 10845817 TI - An analysis of the increased demands placed on dialysis health care team members by functionally dependent hemodialysis patients. AB - A team of health care providers is integral to the care of chronic hemodialysis patients and includes nephrologists, social workers, dietitians, and nurses. Increasingly, the chronic hemodialysis population is composed of older patients with multiple comorbid conditions and reduced functional independence. The demands placed on social workers, nurses, and dietitians caring for the increasingly fragile chronic hemodialysis population have not been examined. We compared the interventions required by social workers, dietitians, and nurses caring for two demographically matched chronic hemodialysis patient groups undergoing dialysis in two outpatient units over a 6-month period to examine the demands imposed by these functionally dependent patients. Patients underwent dialysis in either a step-down unit or an ambulatory unit. Patients undergoing dialysis in the step-down unit had more coronary artery disease (6 of 12 patients [50%] versus 1 of 12 patients; P < 0.025) and peripheral vascular disease (6 of 12 versus 0 patients; P < 0. 004). Mean urea reduction ratio, hematocrit, and serum albumin values, as well as number of hospitalizations and mean days hospitalized for the 6-month study period, were not different between the groups. Patients undergoing dialysis in the step-down unit were more likely to have lower scores on activities of daily living (11 +/- 5 versus 15 +/- 3; P < 0.02), live in a nursing home (58% versus 8%; P < 0.01), be nonambulatory (66% versus 0%; P < 0. 01), and have a catheter as permanent dialysis access (66% versus 9%; P < 0.004). Significantly more social worker and dietitian time in hours per week were provided to the patients in the step-down unit (social workers, 259 versus 201 h/wk; P < 0.001; dietitians, 115 versus 96 h/wk; P < 0.001). Similarly, dialysis treatments requiring nursing interventions (treatments with hypotension, 36% versus 13%; obtaining blood cultures, 7% versus 2%; administering intravenous medications, 9% versus 2%; communicating with other health care providers, 3% versus 0.1%; and non-dialysis-related interventions, 5% versus 0.5%; all P < 0.005) were more common in the patients in the step-down unit. We conclude that increased dialysis provider care is required by patients who are functionally dependent and have increased comorbid conditions. The increased demands this fragile patient population places on dialysis providers must be recognized, examined more closely, and reimbursed appropriately. PMID- 10845818 TI - Spurious metabolic acidosis in hemodialysis patients. AB - Metabolic acidosis with an increased anion gap (AG) is frequently seen among patients with end-stage renal failure that is corrected to a variable degree by chronic hemodialysis. The degree of acidosis is generally interpreted from the concentration of total carbon dioxide (tCO(2)) in blood drawn from the vascular access used for dialysis. As with many dialysis units in the United States, our laboratory studies for outpatients are performed in a central laboratory several hundred miles away and must be shipped there by air freight. We observed a consistent and clinically important difference between the tCO(2) content of samples reported from the central laboratory compared with results reported from a local university hospital chemistry laboratory. The central laboratory readings were always lower, resulting in an increase in the AG. Delays in centrifugation of the blood to separate the clot from the serum and in the initiation of analysis led to an increase in the lactate content of the samples. That increase, however, was insufficient to explain the difference in tCO(2) levels. These data suggest that something happens to the samples in transit to cause an artifactual reduction of the tCO(2) level. For many dialysis patients, the severity of their acidosis may be falsely represented by the tCO(2) content of blood samples reported from central laboratories. PMID- 10845819 TI - Gender difference in the mean age at the induction of hemodialysis in patients with autosomal dominant polycystic kidney disease. AB - Male patients with autosomal dominant polycystic kidney disease (ADPKD) begin hemodialysis earlier than female patients. The rate of progression of many other renal diseases is also faster in men than women. In this study, gender difference in ADPKD was compared with that in other diseases, such as chronic glomerulonephritis, diabetic nephropathy, and nephrosclerosis, using the data obtained from an annual statistical survey of the Japanese Society for Dialysis Therapy. The male-female ratio in ADPKD (n = 8,176) was 1.17:1 and closer to 1.0:1 than the other diseases. Men with ADPKD started hemodialysis therapy 1.3 years earlier than women (male age, 55.9 +/- 12.4 years versus female age, 57.2 +/- 11.5 years), and the age difference was less than that in other diseases. These results suggest that the prognosis in women with ADPKD is relatively worse than that in men with ADPKD or that women are not well protected against the progression of this disease compared with other renal diseases. In conclusion, men with ADPKD are introduced to hemodialysis therapy earlier than women; however, the age difference was small compared with other common renal diseases. PMID- 10845820 TI - ACE inhibitors do not induce recombinant human erythropoietin resistance in hemodialysis patients. AB - Angiotensin-converting enzyme (ACE) inhibitors may exacerbate anemia in patients with chronic renal failure, as well as in dialysis patients. To better answer this question, a prospective, crossover study was conducted to evaluate the effect of ACE inhibitors on recombinant human erythropoietin (rHuEPO) requirements in hemodialysis patients. Patients administered an ACE inhibitor when entering the study remained on this drug for the initial 4 months and were then switched to another antihypertensive agent for 4 more months. Patients not initially administered an ACE inhibitor were switched to lisinopril at 4 months. rHuEPO doses were adjusted using a sliding scale based on weekly laboratory hematocrit values. The inclusion criteria were met by 51 patients undergoing dialysis. Demographics were as follows: 61% were women, 64% were black, 46% had diabetes, average age was 53.2 +/- 13.3 years, and time on hemodialysis was 38.0 +/- 44.5 months. Thirty-three patients completed the study. Hematocrit averaged 32.7% +/- 1.9% while on ACE inhibitor therapy and 33.1% +/- 2.1% off ACE inhibitor therapy (P = 0.217). There was no difference in rHuEPO dose per treatment during each period (3,500 +/- 1,549 U on ACE inhibitor therapy versus 3,312 +/- 1,492 U off ACE inhibitor therapy; P = 0.300). No significant differences were found in degree of blood pressure control or various clinical and laboratory parameters that might be associated with rHuEPO resistance between the two periods. Similarly, no differences were found in hospitalization days, duration of infections, or transfusion requirements. These findings suggest that ACE inhibitors do not contribute to rHuEPO resistance in hemodialysis patients. PMID- 10845821 TI - Hospital-acquired infections among chronic hemodialysis patients. AB - The epidemiological characteristics of nosocomial infections among patients requiring chronic hemodialysis, a high-risk and rapidly growing population, have not been fully elucidated. During a 30-month cohort study, rates of bloodstream infections (BSIs), urinary tract infections (UTIs), pneumonia, and diarrhea caused by Clostridium difficile and the distribution of pathogens among hospitalized chronic hemodialysis patients were compared with hospitalized patients not requiring chronic hemodialysis. To identify risk factors for developing a nosocomial infection among chronic hemodialysis patients, a matched case-control study was performed. A total of 1,557 nosocomial infections were detected during 1,317 of 68,361 admissions (2%). Of these, 47 nosocomial infections occurred in chronic hemodialysis patients during 31 of 578 admissions (5%). Nosocomial infections were significantly more frequent among the chronic hemodialysis group (9.1/1,000 patient-days) compared with the non-chronic hemodialysis group (3. 8/1,000 patient-days; relative risk [RR], 2.4; 95% confidence interval [CI], 1.8 to 3.2; P < 0.001). UTIs were the most common nosocomial infections among chronic hemodialysis patients, accounting for 47% of all infections in this population. UTIs were significantly more common among chronic hemodialysis patients (4.2/1, 000 patient-days) compared with non-chronic hemodialysis patients (0.7/1,000 patient-days; RR, 6.2; 95% CI, 3.8 to 9.5; P < 0.001). Among chronic hemodialysis patients, Candida spp and enterococci were the most common pathogens in contrast to coagulase-negative staphylococci and Staphylococcus aureus among patients not requiring hemodialysis. Using conditional logistic regression, a greater index of comorbidity was significantly associated with nosocomial infections among the chronic hemodialysis population (odds ratio, 3. 6; 95% CI, 1.2 to 10.7; P = 0.02). Chronic hemodialysis patients are at a substantially greater risk for developing a nosocomial infection compared with other hospitalized patients. PMID- 10845822 TI - Accuracy of decrease in blood flow in predicting hemodialysis graft thrombosis. AB - We recently showed that a single low graft blood-flow measurement (Qa) does not accurately predict graft thrombosis. In this study, we prospectively determined whether percentage of decrease in Qa (DeltaQa) or adjustment of Qa for mean arterial pressure (Qa/MAP; Delta(Qa/MAP)) provides greater predictive accuracy than a single Qa. We monitored 83 grafts from 80 patients for thrombosis over periods up to 12 months. Qa (by ultrasound dilution) and MAP were measured monthly during the study. Receiver operating characteristic curves were used to determine whether Qa, DeltaQa, Qa/MAP, or Delta(Qa/MAP) provided the combination of high sensitivity (>80%) and low false-positive rate (FPR; <20%) needed for clinical use. This level of predictive accuracy requires an area under the curve (AUC) of approximately 0.90. We analyzed the four predictors by a number of criteria and found that all AUCs were less than 0.90 and adjustment for MAP reduced the AUC. In predicting thrombosis within 1 month, for example, AUCs for Qa and net DeltaQa (over 3 months) were 0.84 and 0.82, respectively, whereas AUCs for Qa/MAP and net Delta(Qa/MAP) were 0.78 and 0.75, respectively. At a sensitivity of 80%, FPRs for all predictors were at least 30%. Thus, a high sensitivity always required a high FPR. These results show that DeltaQa and adjustment for MAP are not more accurate than a single low Qa in predicting thrombosis. None of these predictors provide enough predictive accuracy to be the sole criterion for clinical decision making. A successful monitoring and intervention program will likely require the inclusion of other predictors that, together with Qa, may provide the needed accuracy. PMID- 10845823 TI - Intradialytic and interdialytic effects of treatment with 1.25 and 1. 75 Mmol/L of calcium dialysate on arterial compliance in patients on hemodialysis. AB - Arterial compliance (AC) is an important determinant of vascular structure, and abnormalities of AC can greatly affect the cardiovascular system. Given the vasoconstrictive properties of increased levels of serum ionized calcium (iCa), we investigated the way that dialysate calcium level can influence AC in the hemodialysis (HD) population. In a crossover randomized design, 19 dialysis patients undergoing regular bicarbonate HD (three times weekly) underwent two cycles of four successive HD sessions each with a low (LdCa; 1.25 mmol/L) and high dialysate calcium concentration (HdCa; 1.75 mmol/L). At the fourth session of each cycle, iCa level and hemodynamic parameters (systolic blood pressure [SBP], diastolic blood pressure, mean arterial pressure [MAP], pulse pressure [PP], heart rate, and AC) were measured pre-HD and post-HD. AC was measured noninvasively at the brachial artery by arterial pulse waveform analysis. The dialysate calcium level was a significant determinant of both pre-HD (r = 0.335; P < 0.05) and post-HD iCa level (r = 0.767; P < 0.001). Pre-HD AC increased significantly (P < 0.05) by 0.01+/- 0.02 mL/mm Hg (7% +/- 19%) on switching from HdCa to LdCa treatment. Multiple regression analysis showed that both pre-HD PP and iCa level were major inverse determinants of pre-HD AC in both the LdCa (R(2) = 0.65; P < 0.001) and HdCa (R(2) = 0.51; P < 0.01) treatment groups. AC increased by 32% (P < 0.01) and 37% (P < 0.05) during LdCa and HdCa dialysis, respectively. Intradialytic changes in AC were inversely correlated with changes in SBP and PP. In the HdCa group, changes in iCa level related significantly to MAP (r = 0.464; P < 0.05). The results show that changes in AC during HD are mainly mediated through concurrent changes of systemic hemodynamics, which are largely affected by dialysate calcium level through parallel changes in iCa level. Interdialytically, a significant, blood pressure-independent, inverse relationship between AC and iCa level exists. Therefore, HD with LdCa, by reducing the incidence of HD-induced hypercalcemia, may have a beneficial role in preventing the ongoing reduction of AC in HD patients and thus improving cardiovascular prognosis. PMID- 10845824 TI - Pattern of noncompliance with dialysis exchanges in peritoneal dialysis patients. AB - The purpose of the study is to evaluate the pattern of noncompliance in peritoneal dialysis (PD) patients using home visit supply inventories. Ninety-two patients were enrolled at the start of dialysis. Noncompliance, defined as performance of less than 90% of prescribed exchanges, was found in 30% of patients during the first 6 months of PD. Patients who were noncompliant with prescribed exchanges at the start of PD had greater rates of death (P = 0.03), transfer to hemodialysis secondary to uremia (P < 0.05), hospitalization (P < 0.001), and days hospitalized (P < 0.001) compared with compliant patients. Delivered Kt/V was 18% less in noncompliant compared with compliant patients (2.1 versus 2.57; P = 0.007). Serial evaluations of compliance in 53 patients showed that 72% were consistently compliant, 2% were consistently noncompliant, 15% were noncompliant at the beginning of PD but became compliant at follow-up, and 11% were intermittently noncompliant. The likelihood of future compliance in a patient compliant at the first home visit was 88%. Patients who were independent with their dialysis exchanges were more likely to be noncompliant (27%) than patients dependent on someone else to perform their dialysis (8%; P = 0.05). Serial 24 hour creatinine excretion was not a useful method to determine compliance. We recommend a home visit during the first 6 months of PD to determine compliance. Those found compliant probably do not need repeated evaluations, whereas noncompliant patients should be reevaluated in a few months. Involving another person in the dialysis might relieve some of the burden on patients who may be initially unable to cope with home dialysis. Identification of noncompliant patients and awareness of risk factors should reduce noncompliance and improve patient outcomes. PMID- 10845825 TI - Tenckhoff catheters prove superior to cook catheters in pediatric acute peritoneal dialysis. AB - Peritoneal dialysis (PD) is the most common form of renal replacement therapy in infants and young children with acute renal failure (ARF). The two most commonly used catheters for performing acute PD are the Cook catheter (CC), placed at the bedside, and the surgically placed Tenckhoff catheter (TC). In the present study, we compared the complications and survival rates of the two catheters. The records of 59 children (age, 1 day to 16.7 years) who underwent PD for ARF from March 1989 through June 1999 in our hospital were reviewed. The initial (primary) catheter was a TC in 22 patients and a CC in 37 patients. The age of the patients who received a primary TC (2.8 +/- 4.5 years) was no different than the age of those with a primary CC (1.4 +/- 2.0 years; P = not significant). The duration of use (mean +/- SD) of TCs (16.5 +/- 14.2 days) was significantly greater than the duration of CC use (4.9 +/- 4.2 days; P < 0.001). Only two patients with a TC (9%) developed complications, whereas 18 patients with a CC (49%) developed complications, 13 of whom required catheter replacement (P < 0.01). Thirty-five patients (59%) recovered renal function after undergoing dialysis for 11.5 +/- 8.0 days. Twenty-three of those patients (66%) required dialysis for more than 5 days. Only 4 patients with a primary CC had successful completion of dialysis without catheter-associated complications compared with 15 patients with a primary TC. Kaplan-Meier survival analysis showed that by day 6 of dialysis, only 46% of primary CCs were functioning without complications compared with 90% of TCs that were free of complications. We conclude that the use of a CC is associated with significantly more complications than a TC, and nearly one half of the CCs are likely to be nonfunctional beyond 5 days of dialysis, at a time when two thirds of the patients are still expected to be undergoing dialysis. Therefore, when possible, a TC should be the catheter of choice when initiating acute PD in children. In those patients for whom a CC is chosen as the initial catheter, an elective change to a TC should be considered once dialysis is expected to extend beyond 5 days. PMID- 10845826 TI - Neutrophil beta(2)-microglobulin and lactoferrin content in renal failure patients. AB - Multiple dysfunctions of polymorphonuclear leukocytes (PMNLs) contribute significantly to the increased morbidity and mortality among patients with end stage renal disease. In the present study, we measured the PMNL content of beta(2)-microglobulin (beta(2)m) and lactoferrin in different states of renal insufficiency and after kidney transplantation. PMNLs were lysed ultrasonically and, after centrifugation, both proteins were assayed in the supernatant by enzyme-linked immunosorbent assay technique. Despite marked differences in plasma beta(2)m levels, no significant difference in PMNL content of beta(2)m and lactoferrin could be shown among the groups analyzed. There was also no correlation between plasma beta(2)m level and PMNL beta(2)m content. In control subjects, as well as in renal allograft recipients with a well-functioning graft, PMNL beta(2)m level correlated positively with PMNL lactoferrin level (pooled data, r = 0.55; P < 0.001; n = 55). Both proteins are considered to colocalize in peroxidase-negative PMNL granules. However, no correlation was found in the azotemic and uremic patient groups. Standard immunofluorescence staining of control PMNLs showed a cytoplasmic granular distribution of both granule proteins. However, in PMNLs of uremic patients, lactoferrin shifted to a perinuclear localization. PMNLs obtained from uremic individuals failed to elicit an increase in lactoferrin release after stimulation with the chemotactic peptide f-Met-Leu-Phe compared with PMNLs obtained from healthy volunteers. These data indicate abnormalities in uremic patients of PMNL granule lactoferrin content and release that are reversible after successful renal transplantation. PMID- 10845827 TI - Mild renal failure is associated with adverse outcome after cardiac valve surgery. AB - The present study was performed to ascertain whether the presence of mild renal failure (defined as a serum creatinine concentration of 1. 5 to 3.0 mg/dL) is an independent risk factor for adverse outcome after cardiac valve surgery. An extensive set of preoperative and postoperative data was collected in 834 prospectively evaluated patients undergoing cardiac valve surgery at 14 Veterans Affairs Medical Centers. Univariate and multivariable analyses were performed to determine whether an independent association of mild renal dysfunction with adverse outcomes was present. Patients with mild renal failure had significantly greater 30-day mortality rates (P = 0.001; 16% versus 6%) and frequency of postoperative bleeding (P = 0.023; 16% versus 8%), respiratory complications (P = 0.02, 29% versus 16%), and cardiac complications (P = 0.002; 18% versus 7%) than patients with normal renal function (serum creatinine <1.5 mg/dL) when controlling for multiple other variables. The presence of a serum creatinine concentration of 1.5 to 3.0 mg/dL is significantly and independently associated with adverse outcomes after cardiac valve surgery. PMID- 10845828 TI - Long-term results with cyclosporine monotherapy in renal transplant patients: a multivariate analysis of risk factors. AB - There is little information on the long-term outcome of patients initially assigned to cyclosporine (CsA) monotherapy and requiring the addition of steroid therapy during follow-up. The aim of this report is to describe our experience with 143 first renal transplant recipients (120 cadaver transplants, 23 living donor transplants) randomized to receive CsA monotherapy as a treatment arm of three consecutive controlled clinical trials. Median follow-up was 86 months. Thirty-four percent of the patients remained on the original CsA monotherapy, whereas the remaining 66% required the addition of steroid therapy. Cumulative patient and graft survivals at 11 years were 0.89 (95% confidence interval [CI], 0.83 to 0.95) and 0.62 (95% CI, 0.52 to 0.72), respectively. The 11-year graft survival for converted patients was 0.53 (95% CI, 0.39 to 0.67). Cumulative graft half-life was 19.9 +/- 3.47 (SE) years. According to the Cox model, variables at transplantation that correlated with a lower 11-year graft survival were yearly increases in age (relative risk [RR], 1. 04; P = 0.039), monthly increases in hemodialysis duration (RR, 1.01; P = 0.029), no blood transfusion before transplantation (RR, 1.99; P = 0.043), CsA administration in a double daily dose (RR, 2.35; P = 0.008), and a cadaver donor transplant (RR, 4.76; P = 0.039). Multivariate analysis of time-dependent variables showed that delayed graft function recovery (RR, 2.20; P = 0.019) and the need to add steroid and/or azathioprine therapy (RR, 5.28; P = 0.000) were also correlated with a lower graft survival. Patients who added steroid therapy developed infections (P < 0.001), cataracts (P < 0.001), cardiovascular complications (P = 0.004), and arterial hypertension (P = 0.024) more frequently than patients remaining on CsA monotherapy. Patients administered CsA in a single daily dose received significantly less CsA over the years (P = 0.0042) than patients administered CsA in two divided doses. They also showed a trend toward greater creatinine clearance levels, although not statistically significant. In conclusion, this analysis showed that in patients assigned to CsA therapy alone, good long-term patient and graft survival probabilities can be obtained. In approximately one third of the patients, the use of steroids could be avoided for up to 11 years, and these patients had a better long-term outcome than those who required the addition of steroid therapy. Finally, in patients administered CsA in a single daily dose, the possibility of reducing CsA dosage probably led to better intrarenal hemodynamics with improving creatinine clearances. PMID- 10845829 TI - Glomerular filtration rate in severely overweight normotensive humans. AB - The study examined whether indexing glomerular filtration rate (GFR) for body surface area is appropriate for people who are severely overweight. Twenty normotensive adult men who were severely overweight but without microalbuminuria were enrolled into this study. The control group consisted of 20 healthy subjects matched for age, sex, and height. GFR was determined by measuring insulin with the continuous-infusion method. The clearance of endogenous creatinine was also measured after two daily urine collections. Renal plasma flow (RPF) was measured by p-aminohippurate clearance using the continuous-infusion method. Lean body weight was measured by impedentiometry. Adjusting for body surface area (in 1.73 m(2)) caused a significant reduction in GFR (P < 0.0001) in overweight humans (84.1 +/- 2.32 versus 109.6 +/- 3.07 mL/min/1.73 m(2)). The difference disappeared when GFR/height criteria were adopted. No difference between obese and healthy controls occurred after adjusting for lean body weight. Data for creatinine clearance paralleled those with insulin clearance; a significant reduction (P < 0.001) occurred after indexing for basal surface area, which disappeared after correction for height, as well as for lean body weight. PMID- 10845830 TI - Nephrotic proteinuria has No net effect on total body protein synthesis: measurements with (13)C valine. AB - In nephrotic syndrome, significant amounts of plasma proteins, mostly of hepatic origin, are lost in urine. Total hepatic protein synthesis increases, suggesting that other protein pools must be conserved to maintain steady state. This can be accomplished either by decreased amino acid oxidation or decreased protein synthesis in other organs to replace lost liver-derived proteins. To determine the effect of nephrotic syndrome on total-body protein metabolism, we compared whole-body valine use in seven nephrotic patients and five controls using a primed continuous infusion of [1-(13)C]-valine, with additional priming of NaH(13)CO(3). Plasma [(13)C]-valine, (13)C alpha ketoisovaleric acid, and the expired (13)CO(2) enrichments were used to assess whole-body valine flux, valine oxidation, and nonoxidative valine disposal (NOVD). The valine flux into the blood compartment (97.7 +/- 3.0 versus 95.3 +/- 3.3 micromol/kg/h), oxidation of valine (19.4 +/- 1.9 versus 21.2 +/- 2. 8 micromol/kg/h), and NOVD (78.3 +/- 2.5 versus 74.2 +/- 2.7 micromol/kg/h) were not statistically different in patients compared with controls. Valine oxidation correlated positively with urinary urea excretion (r = 0.70; P = 0.01) in all subjects. Compared with control subjects who have similar urinary urea excretion, nephrotic subjects do not compensate for urinary loss of protein by decreased amino acid oxidation or decreased nonoxidative valine disposal. Previous studies have shown that synthesis of several hepatic proteins increases when subjects are fed the same dietary regime, whereas the present study shows that total-body protein synthesis does not increase. This would imply reduced synthesis of nonhepatic protein pools. PMID- 10845831 TI - Chronic proteinuric nephropathies: outcomes and response to treatment in a prospective cohort of 352 patients with different patterns of renal injury. AB - The Ramipril Efficacy in Nephropathy (REIN) study found that angiotensin converting enzyme (ACE) inhibitors effectively decreased proteinuria, glomerular filtration rate (GFR) decline (DeltaGFR), and incidence of end-stage renal disease (ESRD) in patients with proteinuric chronic nephropathies. In this study, we prospectively investigated the main clinical determinants of progression and response to treatment in the 352 patients enrolled into the REIN study. Mean DeltaGFR (0.56 +/- 0.05 [SEM] versus 0.21 +/- 0.05 mL/min/1.73 m(2)/mo; P = 0.0001) and incidence of ESRD (30% and 10%; P = 0.0001) were more than twice that in patients with proteinuria of 2 g/24 h or greater of protein compared with those with protein less than 2 g/24 h (relative risk [RR], 4.07; 95% confidence interval [CI], 2.20 to 7.52), as well as in patients with hypertension compared with normotension (mean DeltaGFR, 0.48 +/- 0. 05 versus 0.22 +/- 0.05 mL/min/1.73 m(2)/mon; P = 0.0006; ESRD, 25% versus 10%; P = 0.004; RR, 3.18; 95% CI, 1.38 to 7.32). Hypertension at study entry (P = 0.038), greater mean blood pressure on follow-up (P = 0.002), and urinary protein excretion rate (P = 0.0001) were independent predictors of faster DeltaGFR. DeltaGFR was approximately twofold faster in patients with type 2 diabetes than in those with primary glomerular disease (P = 0.002; including immunoglobulin A [IgA] nephropathy, P = 0.009); nephrosclerosis (P = 0.03), adult polycystic kidney disease (APKD), or chronic interstitial nephritis (P = 0.006). Diabetes at study entry (P = 0. 02) and greater mean blood pressure (P = 0.0001) and urinary protein excretion rate (P = 0.0001) on follow-up were independent predictors of faster DeltaGFR. After correction for baseline covariates, diabetes was also associated with an increased risk for progression to ESRD (RR, 2.39; 95% CI, 1.01 to 5.68; P < 0.05). At multivariate analyses, ramipril significantly decreased DeltaGFR (regression coefficient,-0.23 +/- 0.11 [SEM]; P = 0.036) and ESRD (RR, 2.08; 95% CI, 1.21 to 3.57; P = 0.008) in patients with baseline proteinuria of 2 g/24 h or greater of protein, and the renoprotective effect increased for increasing levels of proteinuria. Ramipril decreased DeltaGFR to a similar extent in normotensive and hypertensive patients (-0.14 +/- 0.11 versus -0.14 +/- 0.09) and significantly limited ESRD in hypertensive patients (RR, 2.03; 95% CI, 1.26 to 3. 26; P = 0.004). DeltaGFR was decreased by 42% in primary glomerular disease (P = 0.017), by 35% in IgA nephropathy, and by 37% in nephrosclerosis, but was not improved in type 2 diabetes, APKD, or interstitial nephritis. At multivariate analyses, ramipril significantly slowed DeltaGFR (-0.24 +/-0.08; P = 0.004) and progression to ESRD (RR, 2.32; 95% CI, 1.36 to 3.96; P = 0.002) in patients without diabetes, but not in patients with diabetes, who tended to have a faster DeltaGFR (+0.62 +/- 0.44) on ramipril therapy. In summary, patients with proteinuria of 2 g/24 h or greater of protein, preexisting hypertension, or type 2 diabetes were faster progressors. Greater blood pressure and degree of proteinuria were the strongest determinants of faster GFR decline. The renoprotective effect of ramipril was similar in patients with normotension and hypertension. Hypertensive patients and those with proteinuria of 2 g/24 h or greater of protein, primary glomerular disease, or nephrosclerosis gained the most from ACE inhibitor treatment. During the study period, those with proteinuria less than 2 g/24 h of protein, type 2 diabetes, or polycystic kidney disease did not benefit by treatment to an appreciable extent. PMID- 10845832 TI - Parvovirus B19 DNA in kidney tissue of patients with focal segmental glomerulosclerosis. AB - Focal segmental glomerulosclerosis (FSGS) represents a clinicopathological syndrome with diverse causes. We examined the possibility that some cases of FSGS are associated with parvovirus B19 infection. We studied renal biopsy tissue from 40 patients, including those with idiopathic FSGS, collapsing FSGS, membranous nephropathy, and minimal change disease, as well as normal renal tissue removed at the time of nephrectomy from 4 patients. DNA was extracted from frozen blocks of kidney tissue and amplified using nested polymerase chain reaction. Parvovirus B19 DNA was amplified from 8 of 10 patients with idiopathic FSGS, 9 of 10 patients with collapsing FSGS, 6 of 10 patients with membranous nephropathy, 5 of 10 patients with minimal change disease, and 2 of 4 cancer nephrectomy samples. The prevalence of parvovirus B19 DNA was greater among patients with idiopathic FSGS and collapsing FSGS compared with patients with other diagnoses (P = 0.05). In situ hybridization studies using digoxigenin-labeled DNA probes failed to detect parvovirus B19 nucleic acid in any of the kidney tissue samples. These results suggest that parvovirus B19 DNA is commonly found in the kidneys of patients with a range of renal diagnoses, possibly representing latent DNA from past infection. The failure to localize parvovirus B19 nucleic acid within kidney argues against ongoing, high-level viral replication. Nevertheless, the increased prevalence of B19 DNA in patients with idiopathic FSGS and collapsing FSGS could indicate a pathogenic role for the virus in the cause of FSGS in certain patients. PMID- 10845833 TI - Protection of alpha(3) integrin-mediated podocyte shape by superoxide dismutase in the puromycin aminonucleoside nephrosis rat. AB - Because reactive oxygen species (ROS) are involved in the development of puromycin aminonucleoside nephrosis (PAN), we examined whether superoxide dismutase (SOD) could ameliorate this condition. Phosphatidyl choline-bound SOD (PC-SOD) has higher affinity for the cell membrane than recombinant human SOD (rhSOD). In this study, PC-SOD had a longer half-life in the circulation and also higher affinity to renal fractions (glomerulus, brush border, and tubulus) than rhSOD. PAN was induced in rats with single injections of puromycin aminonucleoside. Rats were divided into four groups: group P, PAN rats without treatment; group PC-T and group rh-T, PAN rats treated with 30,000 U/kg PC-SOD and rhSOD, respectively; and group C, normal controls. The effect of PC-SOD versus rhSOD on PAN was evaluated by morphological podocyte changes (podocyte density along the GBM) and alpha(3) integrin expression at days 4 and 10. Proteinuria was measured over time until day 14. Distribution and quantitation of alpha(3) integrin were studied by confocal laser scan microscopy. On day 4, glomerular ROS was measured by chemiluminescence without stimulation. PC-SOD decreased proteinuria to the control level, but rhSOD only decreased proteinuria by 31%. PC-SOD significantly improved podocyte density (P < 0.05 versus group P). Total alpha(3) integrin expression decreased in the P and rh-T groups at day 4 and then had recovered by day 10, but the polarity of the site of expression did not recover. PC-T preserved both the amount and polarity of integrin expression on days 4 and 10. PC-SOD significantly suppressed ROS generation in PAN (P < 0.05). These findings suggest that alpha(3) integrin regulates glomerular permeability by maintaining podocyte shape and adhesion, which is disrupted by ROS overproduction. PMID- 10845834 TI - Splenectomy may improve the glomerulopathy of type II mixed cryoglobulinemia. AB - Many patients with type II mixed cryoglobulinemia have been shown to be infected with hapatitis C virus (HCV). Therefore, interferon-alfa has become the first choice of treatment for patients with HCV-associated cryoglobulinemia. However, the disease often relapses after the discontinuation of interferon therapy. The long-term effect of interferon therapy is controversial. Therefore, a more effective therapy needs to be developed. A 62-year-old Japanese woman was admitted to our hospital for the examination of abnormal liver function tests, severe edema, and purpura in her lower extremities. Glomerulopathy secondary to HCV-related cryoglobulinemia was suspected. Her serum creatinine was increased to 2.1 mg/dL. Interferon therapy was considered initially. However, because of pancytopenia caused by liver cirrhosis and splenomegaly, splenectomy was performed in February 1997, before the start of interferon therapy. Renal biopsy specimen taken at the time of the splenectomy showed typical cryoglobulinemic glomerulonephritis. Gradually, after surgery, the patient's thrombocytopenia and anemia improved, her proteinuria and hematuria were decreased, her cryocrit dropped from 15% to 5%, the Ccr increased from 21.1 mL/min to 48.8 mL/min, and the purpura in her lower extremities disappeared. A repeat renal biopsy performed in May 1998 showed marked histological improvement. Splenectomy is not widely accepted as a treatment for cryoglobulinemia. Our case suggests the possibility that the monoclonal-IgM component of the type II cryoglobulin may be formed in the spleen. In conclusion, splenectomy may be an effective therapy for cryoglobulinemia in patients with HCV-positive liver cirrhosis and pancytopenia secondary to splenomegaly. PMID- 10845836 TI - Remission of nephrotic syndrome after removal of localized Castleman's disease. AB - Renal complications of Castleman's disease are uncommon. Among the various renal disorders, including mesangial proliferative glomerulonephritis, membranous glomerulonephritis, and minimal change disease, nephrotic syndrome attributable to renal amyloidosis is very rarely reported. We report a case of mixed type of localized Castleman's disease complicated with nephrotic syndrome. Renal biopsy was performed. The deposition of AA amyloidosis was shown. After the removal of two mesenteric lymphoid masses, the proteinuria was gradually decreased and disappeared. Renal biopsy was repeated after 14 months, and, despite complete remission of nephrotic syndrome, no regression in amyloid deposition was found. PMID- 10845835 TI - "Pauci-Immune" proliferative and necrotizing glomerulonephritis with thrombotic microangiopathy in patients with systemic lupus erythematosus and lupus-like syndrome. AB - In the glomerulonephritides of systemic lupus erythematosus (SLE), the number of subendothelial deposits, when present, generally corresponds to the degree of light microscopic glomerular hypercellularity; only very rarely are no or few such deposits present in cases of focal (WHO class III) or diffuse (WHO class IV) proliferative lupus nephritis. We have recently encountered five cases of active diffuse proliferative glomerlonephritis with no subendothelial and few or no mesangial deposits and thrombotic microangiopathy (TMA) in four patients with SLE and one patient with lupus-like syndrome. Three of the five patients were tested for circulating lupus anticoagulants or anticardiolipin antibodies, and two were positive. All five patients tested negatively for antineutrophil cytoplasmic antibodies (ANCA). Three patients responded to steroid and cyclophosphamide treatment, although one of them died of acute bacterial bronchopneumonia. One patient was lost to follow-up. We conclude that "pauci-immune" proliferative lupus nephritis is rare and should be treated as proliferative lupus nephritis with a proportionate number of subendothelial deposits. The negative ANCA suggests that these cases do not represent incidental ANCA-associated pauci immune necrotizing and crescentic glomerulonephritis in patients with SLE. Of particular interest is that, in patients with SLE, if associated with TMA, an active proliferative necrotizing glomerulonephritis may be present even in the absence of significant glomerular immune complex deposition. PMID- 10845837 TI - Diabetic muscle infarction in patients on dialysis. AB - Spontaneous muscle infarction in diabetic patients is a rare condition that usually occurs in those with advanced diabetic complications. There is a high prevalence of diabetic nephropathy and varying degrees of renal failure. Both type 1 and type 2 diabetics are at risk. The mean age at presentation is approximately 40 years, and both sexes are affected. The cause is uncertain but appears to be attributable to diabetic microangiopathy, with thickening of walls of small arteries and fibrinoid occlusion. There is necrosis of all elements of the muscle, with polymorphonuclear or mononuclear cellular infiltration and a varying but often limited degree of regeneration, depending on the age of the lesion. The presentation is usually acute, with pain and swelling localized to the thigh in most instances. Systemic signs such as pyrexia are infrequent. Laboratory tests (such as white cell count and creatinine kinase) and plain radiographs are not helpful, although the erythrocyte sedimentation rate is often elevated. The diagnosis, in the appropriate setting, is strongly suggested by magnetic resonance imaging, which shows increased signal intensity and asymmetry of the muscle on T2-weighted scanning as well as fluid in the tissue planes. Management consists of resting the muscle, analgesics, and gradual mobilization. Recurrence is common and may be seen in more than 50% of the patients. PMID- 10845838 TI - Poor long-term survival of dialysis patients after acute myocardial infarction: bad treatment or bad disease? PMID- 10845839 TI - Go to bed and get a good night's sleep: you need your rest! PMID- 10845840 TI - Penny wise and bicarbonate foolish. PMID- 10845841 TI - Re-evaluation of risks associated with hyperphosphatemia and hyperparathyroidism in dialysis patients: recommendations for a change in management. AB - Hyperphosphatemia is a predictable consequence of chronic renal failure and is present in most patients on dialysis. Traditionally, the risk associated with elevated serum phosphorus has focused on its impact on renal osteodystrophy. A growing body of evidence, however, suggests that abnormalities in serum phosphorus, calcium-phosphorus product (CaxP), and parathyroid hormone (PTH) levels are resulting in vascular and visceral calcification, thereby contributing to the substantially increased risk of cardiovascular death in this population. In this analysis, we review in detail the literature that describes these associations. We show that the current treatment paradigm for serum phosphorus and secondary hyperparathyroidism is ineffective for a large segment of dialysis patients. Currently, 60% of hemodialysis patients have phosphorus greater than 5.5 mg/dL, and 40% have CaxP greater than 60 mg(2)/dL(2). It is our belief that prevention of uremic calcification, cardiac death, and vascular disease should assume primary importance when evaluating the risks associated with elevated levels of phosphorus, CaxP, and PTH. We recommend that target levels should become 9.2 to 9.6 mg/dL for calcium, 2.5 to 5.5 mg/dL for phosphorus, less than 55 mg(2)/dL(2) for CaxP product, and 100 to 200 pg/mL for intact PTH. PMID- 10845842 TI - Pancreas transplants for diabetic nephropathy: a time for reassessment. PMID- 10845844 TI - Chronic transplant glomerulopathy PMID- 10845843 TI - A 37-year-old woman with systemic lupus erythematosus and acute allograft failure. PMID- 10845845 TI - Continuing medical education exercise, june 2000 PMID- 10845846 TI - Ochrobactrum anthropi bacteremia in a patient on hemodialysis. AB - Although newer tunneled dialysis catheters offer improved capacity for blood flow and efficiency of dialysis, catheter-associated bacteremia remains an extremely important complication of this access strategy. This is a report of a case of catheter-associated bacteremia with Ochrobactrum anthropi, a water-borne gram negative rod with an unusual pattern of antibiotic resistance. Given the organism's hydrophilic property and the frequency of catheter use in debilitated individuals with end-stage renal disease, Ochrobactrum anthropi infection should be considered in the differential diagnosis of a hemodialysis patient with unexplained fever. PMID- 10845847 TI - Acute glomerulonephritis after human parvovirus B19 infection. AB - We evaluated clinical and histological characteristics of four adult patients who presented with acute glomerulonephritic syndrome with serological confirmation of recent HPB19 infection. All patients had generalized edema with urinary abnormalities. Body weight gain ranged from 3 to 10 kg. Three of the patients had contact with erythema infectiosum simultaneously with or within 10 days before development of flu-like symptoms. Two patients had an erythematous rash, and one patient had lower-extremity purpura. Joint pain was present in three of the patients. All patients had proteinuria and hematuria. Renal functions were normal except in one patient who had a serum creatinine of 3.2 mg/dL. Three of the patients had hypocomplementemia. All renal biopsy specimens were characterized by glomerular leukocyte infiltration and endothelial cell swelling. Mesangiolysis was seen in three of the patients. C3 was deposited in a coarse granular pattern along the capillary walls in all cases. Electron microscopic examination showed marked expansion of the subendothelial space of glomerular capillaries in all patients. Subendothelial electron-dense deposits were present in all patients. Immunohistochemical analysis using monoclonal anti-HPB19 antibody showed that one of the four patients had positive staining in the glomeruli. DNA extracted from renal biopsy specimens contained HPB19 DNA, as shown by polymerase chain reaction (PCR) analysis in all patients. PCR amplification of the renal DNA generated a 104-bp product, which hybridized to an HPB19-specific probe. No control group subjects contained HPB19 DNA as determined by PCR. This circumstantial evidence indicates that HPB19 infection may be one of the causes of acute glomerulonephritis in normal individuals. PMID- 10845848 TI - On the adaptation of renal cells to hypertonicity PMID- 10845849 TI - Quest for fire: seeking the source of pathogenic oxygen radicals in atherosclerosis. AB - There is an accumulating body evidence that atherosclerosis is either caused by or accompanied by oxidative events in the vessel wall. These oxidative events have been implicated in proatherogenic modification of proteins, alteration of gene expression, promotion of inflammation, remodeling of vessels, and perturbations of vascular tone. This body of literature has led to a dogma that oxidation is a prerequisite for the atherosclerotic process. In particular, oxidation of lipoproteins by activated macrophages in the subintimal space has been postulated to be an important early step in the atherosclerotic process. PMID- 10845850 TI - Adenoviral vectors and gene transfer to the blood vessel wall. PMID- 10845851 TI - Infection and atherosclerosis: potential roles of pathogen burden and molecular mimicry. AB - Infection has been implicated as a cause of atherosclerosis since the first half of the 19th century. Over the years, sporadic publications have appeared in the literature reflecting a persistent but relatively low level of research activity in this area. In the last decade, however, publications relating to this topic have increased markedly. And very recently, new epidemiological and mechanistic data relating infection to several different diseases, including atherosclerosis, have appeared, stimulating the emergence of important paradigm shifts in how we think about the causes of chronic disease. The following article reviews some of these newer concepts as they relate to a possible role of infection in atherosclerosis. PMID- 10845852 TI - Oxidized lipoproteins and infectious agents: are they in collusion to accelerate atherogenesis? PMID- 10845853 TI - A, B, C...gamma! PMID- 10845854 TI - Viewpoint: are studies in genetically altered mice out of control? AB - Because the use of transgenic and gene-targeted models has increased in popularity, the number of reports describing unpredictable phenotypic effects caused by variation in the genetic background used to generate or propagate these models has steadily increased. There are now many examples in which animals containing the same exact genetic manipulation exhibit profoundly different phenotypes when present on diverse genetic backgrounds, demonstrating that genes unrelated, per se, to the ones being targeted can play a significant role in the observed phenotype. Herein, I will discuss (1) the source of genetic variability in mutant mouse models, (2) the appropriateness of using inbred mice as controls, and (3) strategies to help minimize genetic variation between experimental and control mice. PMID- 10845855 TI - Interactions of oxidants with vascular signaling systems. AB - Individual reactive oxygen species (ROS) and oxidation products of NO interact with vascular signaling mechanisms in ways that appear to have fundamental roles in the control of vascular physiological and pathophysiological function. The activities of ROS-producing systems (including various NADPH and NADH oxidases, xanthine oxidase, and NO synthase) in endothelium and/or vascular smooth muscle are controlled by receptor activation, oxygen tension, metabolic processes, and physiological forces associated with blood pressure and flow. This review focuses on how the chemical properties and metabolic sensing interactions of individual ROS (including superoxide anion, hydrogen peroxide, and peroxynitrite) interact with cellular regulatory systems to produce vascular responses. These species appear to often function through producing selective alterations in individual heme or thiol redox-regulated systems (including guanylate cyclase, cyclooxygenase, mitochondrial electron transport, and tyrosine phosphatases) to initiate physiological responses through signaling pathways that control phospholipases, protein kinases, ion channels, contractile proteins, and gene expression. PMID- 10845856 TI - Development of endothelial cell lines from embryonic stem cells: A tool for studying genetically manipulated endothelial cells in vitro. AB - Totipotent embryonic stem cells can be induced to differentiate to endothelium in vitro. This may be a useful tool for obtaining cultures of genetically manipulated endothelial cells because embryonic stem cells are relatively easy to transfect and are commonly used for gene inactivation experiments in mice. However, embryonic stem cell-derived endothelial cells could not be easily separated from embryoid bodies and maintained in culture. In this study, we describe the isolation and characterization of immortalized endothelial cell lines obtained from embryonic stem cells differentiated in vitro. The cell lines were analyzed for expression of endothelial cell markers, including growth factor receptors and adhesion molecules, and compared with endothelial cells obtained from the yolk sac, the embryo proper, or the heart microcirculation of the adult. We propose that this approach may be useful for obtaining endothelial cells carrying gene mutations that are lethal at very early stages of development. PMID- 10845857 TI - Second-generation adenoviral vectors do not prevent rapid loss of transgene expression and vector DNA from the arterial wall. AB - The utility of adenoviral vectors for arterial gene transfer is limited by the brevity of their expression and by inflammatory host responses. As a step toward circumventing these difficulties, we used a rabbit model of in vivo arterial gene transfer to test 3 second-generation vectors: a vector containing a temperature sensitive mutation in the E2A region, a vector deleted of E2A, and a vector that expresses the immunomodulatory 19-kDa glycoprotein (gp19k) from adenovirus 2. Compared with similar first-generation vectors, the second-generation vectors did not significantly prolong beta-galactosidase transgene expression or decrease inflammation in the artery wall. Although cyclophosphamide ablated the immune and inflammatory responses to adenovirus infusion, it only marginally prolonged transgene expression (94% drop in expression between 3 and 14 days). In experiments performed with "null" adenoviral vectors (no transgene), loss of vector DNA from the arterial wall was also rapid (>99% decrease between 1 hour and 14 days), unrelated to dose, and only marginally blunted by cyclophosphamide. Thus, the early loss of transgene expression after adenoviral arterial gene transfer is due primarily to loss of vector DNA, is not correlated with the presence of local vascular inflammation, and cannot be prevented by use of E2A defective viruses, expression of gp19k, or cyclophosphamide-mediated immunosuppression. Adenovirus-induced vascular inflammation can be prevented by cyclophosphamide treatment or by lowering the dose of infused virus. However, stabilization of adenovirus-mediated transgene expression in the arterial wall is a more elusive goal and will require novel approaches that prevent the early loss of vector DNA. PMID- 10845858 TI - Adenovirus-mediated human tissue kallikrein gene delivery inhibits neointima formation induced by interruption of blood flow in mice. AB - Tissue kallikrein cleaves kininogen to produce vasoactive kinin peptides. Binding of kinins to bradykinin B(2) receptors on vascular endothelial cells stimulates the release of nitric oxide and prostacyclin, thus activating the cGMP and cAMP pathways. In this study, we evaluated the effects of adenovirus-mediated human tissue kallikrein gene (Ad.CMV-cHK) delivery in a mouse model of arterial remodeling induced by permanent alteration in shear stress conditions. Mice underwent ligature of the left common carotid artery and were injected intravenously with saline or 1.8 x 10(9) plaque-forming units of Ad.CMV-cHK or control virus (Ad.CMV-LacZ). Fourteen days after surgery, morphometric analysis revealed that Ad. CMV-cHK reduced neointima formation by 52% (P<0.05) compared with Ad. CMV-LacZ. Expression of human tissue kallikrein (HK) mRNA was detected in mouse carotid artery, aorta, kidney, heart, and liver, and recombinant HK was present in the urine and plasma of mice receiving HK gene. Kallikrein gene transfer resulted in increases in urinary kinin, cGMP, and cAMP levels. The protective action of Ad. CMV-cHK on neointima formation was significantly reduced (P<0.05) in mice with knockout of the kinin B(2) receptor gene compared with wild type control mice (J129Sv mice). In contrast, the effect of Ad. CMV-cHK was amplified (P<0.05) in transgenic mice overexpressing human B(2) receptor compared with wild-type control mice (c57/Bl6 mice). Thus, the inhibitory effect of recombinant kallikrein on structural alterations caused by the interruption of blood flow appears to be mediated by the B(2) receptor. These results provide new insight into the role of the tissue kallikrein-kinin system in vascular remodeling and suggest the application of HK gene therapy to treat restenosis and atherosclerosis. PMID- 10845859 TI - Smooth muscle cell matrix metalloproteinase production is stimulated via alpha(v)beta(3) integrin. AB - This study tests the hypothesis that alpha(v)beta(3) integrin receptors play a critical role in smooth muscle cell (SMC) migration after arterial injury and facilitate migration through the upregulation of matrix metalloproteinase (MMP) activity. We showed that beta(3) integrin mRNA was upregulated by SMCs in the balloon-injured rat carotid artery in coincidence with MMP-1 expression and early SMC migration. Treatment with the beta(3) integrin-blocking antibody F11 significantly decreased SMC migration into the intima at 4 days after injury, from 110.8+/-30.8 cells/mm(2) in control rats to 10.29+/-7.03 cells/mm(2) in F11 treated rats (P=0.008). By contrast, there was no effect on medial SMC proliferation or on medial SMC number in the carotid artery at 4 days. In vitro, we found that human newborn SMCs produced MMP-1 but that adult SMCs did not. This was possibly due to the fact that newborn SMCs expressed alpha(v)beta(3) integrin receptors, whereas adult SMCs did not. Stimulation of newborn (alpha(v)beta(3)+) SMCs with osteopontin, a matrix ligand for alpha(v)beta(3), increased MMP-1 production from 114.4+/-35.8 ng/mL at 0 nmol/L osteopontin to 232.5+/-57.5 ng/mL at 100 nmol/L osteopontin. Finally, we showed that stimulation of newborn SMCs with platelet-derived growth factor-BB and osteopontin together increased the SMC production of MMP-9. Thus, our results support the hypothesis that SMC alpha(v)beta(3) integrin receptors play an important role in regulating migration by stimulating SMC MMP production. PMID- 10845860 TI - Enhanced H(2)O(2)-induced cytotoxicity in "epithelioid" smooth muscle cells: implications for neointimal regression. AB - Vascular smooth muscle cells (SMCs) are phenotypically diverse. Although most medial SMCs can be classified as "fusiform," others are of the "epithelioid" phenotype. Proliferation and apoptosis of epithelioid SMCs may contribute importantly to neointimal formation and regression, respectively. Because reactive oxygen species (ROS) are increased in vascular injury and can induce apoptosis of SMCs, we compared the effects of ROS on epithelioid and fusiform SMCs. Epithelioid and fusiform SMC lines were clonally isolated from rat aortic media and studied under similar conditions and passage numbers. H(2)O(2) produced dose- and time-dependent cytotoxicity that was enhanced in epithelioid compared with fusiform cells. After 24-hour exposure to 50 micromol/L H(2)O(2), epithelioid cell numbers were reduced by 34+/-5% versus a 3+/-5% (P<0.05) reduction in fusiform cell numbers. Similar results were obtained whether H(2)O(2) was administered to growth-arrested or growing cells or when epithelioid and fusiform cells were exposed to extracellular O(2)(.-). To investigate whether apoptosis contributed to enhanced ROS-induced cytotoxicity in epithelioid SMCs, terminal deoxyribonucleotidyl transferase (TDT)-mediated dUTP-digoxigenin nick end labeling (TUNEL) staining was performed. The incidence of TUNEL positivity was 5-fold increased in epithelioid versus fusiform SMCs after treatment with 50 micromol/L H(2)O(2) (19+/-1% epithelioid versus 5+/-1% fusiform, P<0.05). Enhanced H(2)O(2)-induced apoptosis in epithelioid SMCs was confirmed by DNA laddering. Furthermore, when balloon-injured aortas were exposed to H(2)O(2) ex vivo, enhanced apoptosis was observed in neointimal compared with medial SMCs. These results suggest that epithelioid SMCs exhibit enhanced sensitivity to ROS induced apoptosis, which may play an important role in neointimal regression. PMID- 10845861 TI - Increased hyaluronan and hyaluronidase production and hyaluronan degradation in injured aorta of insulin-resistant rats. AB - Diabetic patients have a greater incidence of restenosis, which has been shown to be related to exaggerated intimal hyperplasia. Hyaluronan (HA) has been shown to be closely involved in arterial smooth muscle cell proliferation and migration, which provoke intimal hyperplasia after balloon catheter injury. Our aim was to determine the effect of fructose feeding, which produces certain characteristics of non-insulin-dependent diabetes (ie, insulin resistance, hyperinsulinemia, and hypertriglyceridemia), on production of HA and hyaluronidase and degradation of HA in rat aorta. Treated rats received fructose (25% in tap water) 12 weeks before balloon catheter injury and 14 days afterward. Fructose-fed rats had hyperinsulinemia and hypertriglyceridemia. Injury increased intima-media wet weight (7.5%) and DNA content (20%) in control rats. This increase was significantly greater in fructose-fed rats (22% for wet weight and 34% for DNA content) and was associated with greater HA and hyaluronidase production (123% and 41%, respectively) than in control rats (49% and 7%, respectively). Determination of HA molecular mass showed that balloon catheter injury increased the number of HA fragments in the aorta of control rats. Normal aorta of fructose fed rats contained more HA fragments than that of control rats. Injury to the aorta of fructose-fed rats increased HA fragments and induced the appearance of a very-high-molecular-mass (>2000 kDa) HA. In conclusion, fructose treatment, which induced hyperinsulinemia and hypertriglyceridemia, increased HA and hyaluronidase production and HA degradation in injured aorta. This finding suggests that HA, which has been shown to play a crucial role in proliferation and migration of arterial smooth muscle cells, may be involved in the promotional effect of long term fructose feeding on arterial wall reaction to injury. PMID- 10845862 TI - Alpha(2)-antiplasmin gene deficiency in mice does not affect neointima formation after vascular injury. AB - The hypothesis that alpha(2)-antiplasmin (alpha(2)-AP), the main physiological plasmin inhibitor, plays a role in neointima formation was tested with use of a vascular injury model in wild-type (alpha(2)-AP(+/+)) and alpha(2)-AP-deficient (alpha(2)-AP(-/-)) mice. The neointimal and medial areas were similar 1 to 3 weeks after electric injury of the femoral artery in alpha(2)-AP(+/+) and alpha(2)-AP(-/-) mice, resulting in comparable intima/media ratios (eg, 0.43+/ 0.12 and 0.42+/-0.11 2 weeks after injury). Nuclear cell counts in cross sectional areas of the intima of the injured region were also comparable in arteries from alpha(2)-AP(+/+) and alpha(2)-AP(-/-) mice (78+/-19 and 69+/-8). Fibrin deposition was not significantly different in arteries of both genotypes 1 day after injury, and no mural thrombosis was detected 1 week after injury. Fibrinolytic activity in femoral arterial sections, as monitored by fibrin zymography, was higher in alpha(2)-AP(-/-) mice 1 week after injury (P<0.001) but was comparable in both genotypes 2 and 3 weeks after injury. Staining for elastin did not reveal significant degradation of the internal elastica lamina in either genotype. Immunocytochemical analysis revealed a comparable distribution pattern of alpha-actin-positive smooth muscle cells in both genotypes. These findings indicate that the endogenous fibrinolytic system of alpha(2)-AP(+/+) mice is capable of preventing fibrin deposition after vascular injury and suggest that alpha(2)-AP does not play a major role in smooth muscle cell migration and neointima formation in vivo. PMID- 10845863 TI - Peripheral vascular endothelial dysfunction and apoptosis in old monkeys. AB - To determine the effects of aging on vasoactivity in a primate model (Macaca fascicularis), 13 young male monkeys (aged 7.1+/-0.4 years) and 9 old male monkeys (aged 19.8+/-0.6 years) were chronically instrumented for measurement of left ventricular and aortic pressures and cardiac output. Total cholesterol, triglyceride, and fasting blood sugar levels were not different between the 2 groups. There were no significant differences in baseline mean aortic pressure and total peripheral resistance (TPR) in the young monkeys versus the old monkeys. TPR fell less (P<0.05) with acetylcholine (1 microg/kg) in old monkeys ( 25+/-1%) than in young monkeys (-34+/-2%), whereas decreases in TPR with sodium nitroprusside were similar in old and young monkeys. There was no evidence of atherosclerosis, but apoptosis of endothelial cells was enhanced (P<0.05) in the aortas and femoral arteries, but not in the media, of the old monkeys. There was a relationship (r=0.62, P=0.013) between the incidence of terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL)-positive endothelial cells and endothelial cell density in the femoral artery. The reduced endothelial cell density was also correlated (r=0.82, P<0.01) with depressed TPR responses to acetylcholine. Thus, vascular endothelial dysfunction was present in old monkeys without evidence of atherosclerosis, which may be due to endothelial apoptosis and reduced endothelial cell density. PMID- 10845864 TI - Phenotypic modulation by fibronectin enhances the angiotensin II-generating system in cultured vascular smooth muscle cells. AB - We previously demonstrated that homogeneous cultures of vascular smooth muscle cells (VSMCs) from spontaneously hypertensive rats produce angiotensin II (Ang II) in response to increases in the levels of angiotensinogen, cathepsin D, and angiotensin-converting enzyme (ACE). The change of VSMCs from the contractile to the synthetic phenotype increased the amount of synthetic organelles, resulting in the production of proteases and growth factors. To evaluate the contribution of the synthetic phenotype to the generation of Ang II, we examined the effect of fibronectin (FN), which reportedly induces the synthetic phenotype, on the Ang II generating system in VSMCs. Cultured VSMCs from Wistar-Kyoto rats were incubated with an active fragment of FN, Arg-Gly-Asp-Ser, for 24, 48, or 72 hours after synchronization of the cell cycle with 0. 2% calf serum for 48 hours. Immunofluorescence and protein levels of alpha-smooth muscle (SM) actin and expression of SM22alpha mRNA, apparent in the contractile phenotype, were suppressed by FN, whereas expression of matrix Gla mRNA and osteopontin mRNA and protein, apparent in the synthetic phenotype, was increased. FN (1 to 1000 microg/mL) dose-dependently increased DNA synthesis in the VSMCs, which was inhibited by the Ang II type 1 receptor antagonist CV-11974. Ang II-like immunoreactivity as determined by radioimmunoassay was significantly increased in conditioned medium from the VSMCs. In addition, mRNA for the Ang II-generating proteases cathepsin D and ACE was increased by FN. Expression of transforming growth factor-beta1, platelet-derived growth factor A-chain, and basic fibroblast growth factor mRNAs was also increased by FN. These results indicate that the changes accompanying the alteration to the synthetic phenotype in homogeneous cultures of VSMCs increase expression of proteases such as cathepsin D and ACE, which then produce Ang II, and that these changes increase expression of growth factors that then induce growth of VSMCs. PMID- 10845865 TI - Nitric oxide enhances expression and shedding of tumor necrosis factor receptor I (p55) in endothelial cells. AB - The biological actions of tumor necrosis factor-alpha (TNF-alpha) are mediated by 2 distinct receptors, TNF-RI (p55) and TNF-RII (p75). The extracellular domains of both receptors are shed in soluble form (sTNF-RI and sTNF-RII). The soluble receptors are involved in regulating TNF-alpha activities and may have therapeutic potential as TNF-neutralizing agents. However, it remains unclear as to what kind of physiological molecule can regulate TNF receptors. Nitric oxide (NO) mediates a variety of biological and pathophysiological functions. We hypothesized that NO may modulate the expression and shedding of TNF-RI. An NO donor, diethylamine/NO complex (NOC 5), increased sTNF-RI in the supernatants of ECV304, a human umbilical vein cell line, in a dose-dependent manner. TNF-RI mRNA in these cells was upregulated by NOC 5. 8-Br-cGMP and peroxynitrate had no effect on sTNF-RI release. Genistein and herbimycin A, inhibitors of tyrosine kinase, inhibited sTNF-RI release. Herbimycin A inhibited the levels of TNF-RI mRNA enhanced by NOC 5, which downregulated the surface expression of TNF-RI, indicating that NO is also involved in the shedding process of TNF-RI. The shedding of TNF-RI was abolished by a synthetic inhibitor of matrix metalloproteinase, KB-R8301. In conclusion, NO enhanced the release of sTNF-RI from endothelial cells by a cGMP-independent mechanism. Dual pathways suggested for NO-induced sTNF-RI release include (1) enhanced expression of TNF-RI, at least partially, by a tyrosine kinase-dependent mechanism and (2) increased shedding of TNF-RI by a type of metalloproteinase. PMID- 10845866 TI - Vascular protection: A novel nonangiogenic cardiovascular role for vascular endothelial growth factor. AB - There is widespread interest in the use of the angiogenic cytokine, vascular endothelial growth factor (VEGF), for the treatment of cardiovascular disease. The main paradigm for VEGF cardiovascular therapy is the stimulation of "therapeutic angiogenesis" in ischemic myocardial and peripheral vascular limb disease. In this review, approaches to VEGF therapy based on the therapeutic angiogenesis model are critically assessed, and the alternative mechanism of vascular protection is advanced. Vascular protection is defined as the VEGF induced enhancement of endothelial functions that mediate the inhibition of vascular smooth muscle cell proliferation, enhanced endothelial cell survival, suppression of thrombosis, and anti-inflammatory effects. VEGF-induced synthesis of NO and prostacyclin are both likely to be key mediators of VEGF-dependent vascular protection. Investigation into vascular protection should help us to gain insight into the underlying mechanisms of the cardiovascular actions of VEGF and should prove valuable in the development of novel therapeutic approaches based on local VEGF gene delivery. PMID- 10845867 TI - Lipoteichoic acid induces delayed protection in the rat heart: A comparison with endotoxin. AB - Classic ischemic preconditioning transiently (30 to 120 minutes) protects the myocardium against subsequent lethal ischemia/reperfusion injury. After dissipation of this acute protection, a second window of protection (SWOP) appears 12 to 24 hours later; this SWOP lasts up to 3 days. Several triggers induce a SWOP, including brief repetitive cycles of coronary artery occlusion, rapid ventricular pacing, stimulation of adenosine A(1) receptors, and administration of wall fragments of Gram-negative bacteria, such as lipopolysaccharide (LPS). The aim of this study was to investigate whether lipoteichoic acid (LTA), a cell wall fragment of Gram-positive bacteria, can induce a SWOP in a rat model of left anterior descending coronary artery (LAD) occlusion (25 minutes) and reperfusion (2 hours). Thus, 166 male Wistar rats were pretreated (2 to 24 hours) with saline, LTA (1 mg/kg IP), or LPS (1 mg/kg IP) and subjected to LAD occlusion/reperfusion. Pretreatment with LTA or LPS for 16 hours led to a substantial, approximately 65%, reduction in infarct size and a reduction in the release of cardiac troponin T into the plasma. The dose of LTA used had no toxic effect (on any of the parameters studied), whereas the same dose of LPS caused a time-dependent activation of the coagulation system and liver injury. By use of RNase protection assays, it was determined that LPS caused a time-dependent induction of tumor necrosis factor-alpha, interleukin 1beta, and manganese superoxide dismutase mRNA content in the heart, whereas LTA failed to induce manganese superoxide dismutase. LPS also caused an upregulation of the expression of intercellular adhesion molecule-1 and P-selectin, whereas LTA downregulated these molecules and attenuated the accumulation of polymorphonuclear granulocytes caused by myocardial ischemia/reperfusion. This study demonstrates for the first time that pretreatment with LTA at 8 to 24 hours before myocardial ischemia significantly reduces (1) infarct size, (2) cardiac troponin T, and (3) the histological signs of tissue injury in rats subjected to LAD occlusion and reperfusion. The mechanism(s) underlying the observed cardioprotective effects of LTA warrants further investigation but is likely to be related to its ability to inhibit the interactions between the coronary vascular endothelium and polymorphonuclear granulocytes. Therefore, LTA represents a novel and promising agent capable of enhancing myocardial tolerance to ischemia/reperfusion injury. PMID- 10845868 TI - Impaired superoxide production due to a deficiency in phagocyte NADPH oxidase fails to inhibit atherosclerosis in mice. AB - Superoxide, the reduced form of molecular oxygen, has been implicated in the genesis of vascular disease. One potential mechanism involves oxidation of low density lipoprotein into an atherogenic particle. A second involves reaction with nitric oxide to generate peroxynitrite, a highly oxidizing intermediate. A third involves regulation of signal transduction in artery wall cells. One well characterized pathway for superoxide production resides in macrophages, the cellular hallmark of the early atherosclerotic lesion. Macrophages contain a membrane-bound NADPH oxidase that reduces oxygen to superoxide. In the current studies, we used mice that are deficient in the gp91-phox subunit of the NADPH oxidase-a model of chronic granulomatous disease (CGD)-to explore the role of superoxide in atherosclerotic vascular disease. Wild-type and CGD mice on the C57BL/6 background received a high-fat diet for 20 weeks to induce hypercholesterolemia. At the end of this period, the 2 strains of mice had comparable plasma lipid levels, and their atherosclerotic lesions were similar in size. We also crossed CGD mice with apolipoprotein E-deficient (apoE-/-) mice to generate spontaneously hypercholesterolemic animals that lacked functional NADPH oxidase. After 24 weeks, the CGD-apoE-/- animals had lower plasma cholesterol and triglyceride levels than did the apoE-/- animals, but there was no difference in the extent of atherosclerotic plaque. Our findings suggest that superoxide generated by the NADPH oxidase of phagocytes does not promote atherosclerosis in mice with either diet-induced or genetic forms of hypercholesterolemia. PMID- 10845869 TI - Infection and inflammation induce LDL oxidation in vivo. AB - Epidemiological studies have shown an increased incidence of coronary artery disease in patients with chronic infections and inflammatory disorders. Because oxidative modification of lipoproteins plays a major role in atherosclerosis, the present study was designed to test the hypothesis that the host response to infection and inflammation induces lipoprotein oxidation in vivo. Lipoprotein oxidation was measured in 3 distinct models of infection and inflammation. Syrian hamsters were injected with bacterial lipopolysaccharide (LPS), zymosan, or turpentine to mimic acute infection, acute systemic inflammation, and acute localized inflammation, respectively. Levels of oxidized fatty acids in serum and lipoprotein fractions were measured by determining levels of conjugated dienes, thiobarbituric acid-reactive substances, and lipid hydroperoxides. Our results demonstrate a significant increase in conjugated dienes and thiobarbituric acid reactive substances in serum in all 3 models. Moreover, LPS and zymosan produced a 4-fold to 6-fold increase in conjugated diene and lipid hydroperoxide levels in LDL fraction. LPS also produced a 17-fold increase in LDL content of lysophosphatidylcholine that is formed during the oxidative modification of LDL. Finally, LDL isolated from animals treated with LPS was significantly more susceptible to ex vivo oxidation with copper than LDL isolated from saline treated animals, and a 3-fold decrease occurred in the lag phase of oxidation. These results demonstrate that the host response to infection and inflammation increases oxidized lipids in serum and induces LDL oxidation in vivo. Increased LDL oxidation during infection and inflammation may promote atherogenesis and could be a mechanism for increased incidence of coronary artery disease in patients with chronic infections and inflammatory disorders. PMID- 10845870 TI - Expression of mouse acute-phase (SAA1.1) and constitutive (SAA4) serum amyloid A isotypes: influence on lipoprotein profiles. AB - The serum amyloid A (SAA) family of proteins consists of inducible acute-phase members and a constitutive member that are minor apolipoproteins of normal high density lipoprotein (HDL). During inflammation, HDL cholesterol and apolipoprotein A-I (apoA-I) protein are decreased, and these changes are thought to be partly related to the increase in acute-phase SAA proteins that associate with the HDL particle to become the major apolipoprotein species. To determine the specific role of SAA in the alteration of HDL in the absence of a generalized acute-phase response, acute-phase Saa1.1 transgene expression was directed via an inducible mouse metallothionein promoter. Elevated levels of SAA1.1 (28+/-9 mg/dL) comparable to a moderate acute-phase response were achieved over a 5-day period. SAA association with the HDL particles at this concentration did not significantly alter the apoA-I or HDL cholesterol levels or change the lipoprotein profiles in the transgenic mice compared with wild-type mice. In addition, we used adenoviral vectors to increase the SAA expression to levels seen in a major acute-phase response. Injection of adenovirus expressing the mouse SAA1.1 protein resulted in high-level expression (72+/-8 mg/dL) but did not alter apoA-I levels. However, the SAA associated with the HDL particle gave rise to significantly larger HDL particles ( approximately 10%). Adenoviral expression of the constitutive SAA4 protein resulted in an increase in HDL size ( approximately 10%) and an increase in very low density lipoprotein levels (20 fold) and triglyceride levels (1.7-fold). These studies suggest that increases in acute-phase SAA proteins alone are insufficient to alter HDL cholesterol or apoA I levels during inflammation. A role for constitutive SAA4 in HDL-very low density lipoprotein interactions should be considered. PMID- 10845871 TI - Cafestol increases serum cholesterol levels in apolipoprotein E*3-Leiden transgenic mice by suppression of bile acid synthesis. AB - Cafestol, a diterpene present in unfiltered coffee, potently increases serum cholesterol levels in humans. So far, no suitable animal model has been found to study the biochemical background of this effect. We determined the effect of cafestol on serum cholesterol and triglycerides in different mouse strains and subsequently studied its mechanism of action in apolipoprotein (apo) E*3-Leiden transgenic mice. ApoE*3-Leiden, heterozygous low density lipoprotein-receptor (LDLR+/-) knockout, or wild-type (WT) C57BL/6 mice were fed a high- (0.05% wt/wt) or a low- (0.01% wt/wt) cafestol diet or a placebo diet for 8 weeks. Standardized to energy intake, these amounts are equal to 40, 8, or 0 cups of unfiltered coffee per 10 MJ per day in humans. In apoE*3-Leiden mice, serum cholesterol was statistically significantly increased by 33% on the low- and by 61% on the high cafestol diet. In LDLR+/- and WT mice, the increases were 20% and 24%, respectively, on the low-cafestol diet and 55% and 46%, respectively, on the high cafestol diet. These increases were mainly due to a rise in very low density lipoprotein (VLDL) and intermediate density lipoprotein cholesterol in all 3 mouse strains. To investigate the mechanism of this effect, apoE*3-Leiden mice were fed a high-cafestol or a placebo diet for 3 weeks. Cafestol suppressed enzyme activity and mRNA levels of cholesterol 7alpha-hydroxylase by 57% and 58%, respectively. mRNA levels of enzymes involved in the alternate pathway of bile acid synthesis, ie, sterol 27-hydroxylase and oxysterol 7alpha-hydroxylase, were reduced by 32% and 48%, respectively. The total fecal bile acid output was decreased by 41%. Cafestol did not affect hepatic free and esterified cholesterol, but it decreased LDLR mRNA levels by 37%. The VLDL apoB and triglyceride production rates, as measured after Triton injection, were 2-fold decreased by cafestol, indicating that the number of particles secreted had declined and that there was no change in the amount of triglycerides present in the VLDL particle during cafestol treatment. However, the VLDL particles contained a 4-times higher amount of cholesteryl esters, resulting in a net 2 fold increased secretion of cholesteryl esters. The decrease in triglyceride production was the result of a reduction in hepatic triglyceride content by 52%. In conclusion, cafestol increases serum cholesterol levels in apoE*3-Leiden mice by suppression of the major regulatory enzymes in the bile acid synthesis pathways, leading to decreased LDLR mRNA levels and increased secretion of hepatic cholesterol esters. We suggest that suppression of bile acid synthesis may provide an explanation for the cholesterol-raising effect of cafestol in humans. PMID- 10845872 TI - Plasma concentration of asymmetric dimethylarginine, an endogenous inhibitor of nitric oxide synthase, is elevated in monkeys with hyperhomocyst(e)inemia or hypercholesterolemia. AB - Hyperhomocyst(e)inemia is associated with endothelial dysfunction. Mechanisms responsible for endothelial dysfunction in hyperhomocyst(e)inemia may involve impaired bioavailability of endothelium-dependent nitric oxide. We tested the hypothesis that hyperhomocyst(e)inemia is associated with an elevated plasma concentration of asymmetric dimethylarginine (ADMA), an endogenous inhibitor of nitric oxide synthase. One group of adult cynomolgus monkeys was fed either a control or hyperhomocyst(e)inemic diet for 4 weeks in a randomized crossover design. The second group was fed an atherogenic diet that produces both hyperhomocyst(e)inemia and hypercholesterolemia for 17 months, followed by an atherogenic diet supplemented with B vitamins for 6 months to decrease plasma homocyst(e)ine concentration. Human endothelial cells were used to study the effects of methionine and homocysteine in the presence or absence of B vitamins or the methylation inhibitor S-adenosylhomocysteine on the formation of ADMA and its inactive stereoisomer, symmetric dimethylarginine. The hyperhomocyst(e)inemic diet produced 2- to 3-fold increases in plasma levels of homocyst(e)ine and ADMA (both P<0.05). The atherogenic diet also produced elevated plasma levels of homocyst(e)ine and ADMA (both P<0. 05). Supplementation of the atherogenic diet with B vitamins decreased the plasma levels of homocyst(e)ine but did not affect the plasma levels of ADMA or endothelial function. There was a strong correlation between plasma ADMA and homocyst(e)ine and a strong inverse correlation between ADMA and carotid artery relaxation to acetylcholine. ADMA release by cultured endothelial cells was significantly increased in the presence of methionine or homocysteine. This effect was blocked by S-adenosylhomocysteine but not by B vitamins. We conclude that plasma levels of ADMA are elevated in hyperhomocyst(e)inemia. Because ADMA acts as a competitive inhibitor of endothelial nitric oxide synthase, these findings suggest a novel mechanism for impaired endothelial function in hyperhomocyst(e)inemia. PMID- 10845873 TI - Interferon-gamma induces downregulation of Tangier disease gene (ATP-binding cassette transporter 1) in macrophage-derived foam cells. AB - Cholesterol efflux is a fundamental process that serves to mitigate cholesterol accumulation and macrophage foam cell formation. Recently, we reported that cholesterol efflux to high density lipoprotein subfraction 3 was reduced by interferon-gamma (IFN-gamma) and that this decrease was associated with an increase in acyl coenzyme A:cholesterol acyltransferase (ACAT) expression. In the present study, although treatment of murine peritoneal macrophages with IFN-gamma resulted in a 2-fold decrease in HDL-mediated cholesterol efflux, efflux to lipid free apolipoprotein A-I was reduced >4-fold and approached basal levels. This decrease was associated with a 3- to 4-fold reduction in ATP-binding-cassette transporter 1 (ABC1) mRNA content, the gene responsible for the defect in Tangier disease. Consistent with the reduction in cholesterol and phospholipid efflux in Tangier fibroblasts, downregulation of ABC1 expression by IFN-gamma also resulted in reduced phosphatidylcholine and sphingomyelin efflux to apolipoprotein A-I. Whereas foam cells had a 3-fold increase in ABC1 mRNA, the decrease in ABC1 message levels by IFN-gamma was observed in foam cells and control macrophages. This effect of IFN-gamma was independent of general macrophage activation (inasmuch as similar changes were not detected with granulocyte-macrophage colony stimulating factor) and was not observed with other ABC transporters (inasmuch as the expression of the transporter in antigen processing was upregulated 4-fold in these same cells). Therefore, by decreasing cholesterol efflux through pathways that include the upregulation of ACAT and the downregulation of ABC1, IFN-gamma can shift the equilibrium between macrophages and foam cells and thus impact the progression of an atherosclerotic lesion. PMID- 10845874 TI - LDL receptor-related protein mediates uptake of aggregated LDL in human vascular smooth muscle cells. AB - Foam cell formation is a key event in the onset and progression of atherosclerotic lesions. We have previously reported that internalization of aggregated low density lipoproteins (agLDLs) by vascular smooth muscle cells (VSMCs) produces cholesteryl ester (CE) accumulation in these cells. The aim of this study was to analyze whether the low density lipoprotein receptor-related protein (LRP) mediates the uptake of agLDL by VSMCs. First, immunocytochemistry and fluorescence microscopic analysis with the use of anti-LRP antibodies indicated that there was a high expression of LRP in VSMCs. Confocal microscopic analysis with the use of agLDLs labeled with fluorochrome 1,1'-dioctadecyl 3,3,3', 3'-tetramethylindocarbocyanine and anti-LRP antibodies showed the colocalization of agLDL and LRP. The second approach was to analyze the effect of LRP ligands on agLDL internalization; lactoferrin strongly inhibited CE accumulation from agLDLs (85.0+/-5.7% at 25 microg/mL) by impairing agLDL binding. Coincubation of agLDL with anti-LRP antibodies decreased in a dose dependent manner agLDL-derived CE accumulation (from 20% at 12.5 microg/mL to 80% at 50 microg/mL). The third approach was to evaluate whether antisense LRP oligodeoxynucleotides were able to block agLDL internalization. Treatment of VSMCs with 5 micromol/L antisense LRP oligodeoxynucleotides reduced agLDL-derived CE accumulation by 84+/-2%. In conclusion, these results from immunologic, biochemical, and molecular interventions demonstrate that LRP mediates the binding and internalization of agLDL in human VSMCs. Because LRP is highly expressed in VSMCs and the uptake of 1 LDL aggregate amounts to the deposition of several hundreds of LDL particles, the uptake of agLDL through LRP could have a crucial role for lipid deposition in VSMCs. PMID- 10845875 TI - Differential effect of National Cholesterol Education Program (NCEP) Step II diet on HDL cholesterol, its subfractions, and apoprotein A-I levels in hypercholesterolemic women and men after 1 year: the beFIT Study. AB - We previously reported that high density lipoprotein cholesterol (HDL-C) decreases more in hypercholesterolemic (HC) women than in HC men ingesting an National Cholesterol Education Program (NCEP) Step II diet for 6 months. We examined these subjects to determine whether the differential HDL-C reduction persists after 12 months and whether it is associated with decreased HDL(2)-C and apoprotein A-I. Subjects were screened from an industrial workforce and were defined as HC if 2 low density lipoprotein cholesterol measurements were >/=75th percentile or defined as combined hyperlipidemic (CHL) if triglycerides were also >/=75th percentile. The subjects were then taught the NCEP Step II diet in 8 weekly classes and counseled quarterly. Seventy-three HC and 92 CHL women (mean ages 43 and 44 years, respectively) and 112 HC and 106 CHL men (ages 45 and 41 years, respectively) were studied. All groups reported similar total fat (24% to 26% kcal) and saturated fat (7.1% to 7.9% kcal) intakes at 1 year. HDL-C decreased 7.6% in HC women (P<0.01), exceeding the nonsignificant 1.3% decrease in HC men (P=0.000). HDL(2)-C decreased 16.7% in HC women (P<0.01) compared with the nonsignificant 0.5% increase in HC men (P=0.000). In CHL women and men, HDL-C decreased 3.5% and 3.9% (both P<0.01); HDL(2)-C decreased more in women (7.1%, P<0.01) than in men (4.3%, a nonsignificant difference). Apoprotein A-I decreased significantly (5.3%, P<0.01) in HC women only. Plasma triglycerides were unchanged. Low density lipoprotein cholesterol and weight changes were not different among the 4 groups. HDL-C, HDL(2)-C, and apoprotein A-I levels decreased more in HC women than in HC men after following the NCEP Step II diet for 1 year, continuing a trend observed with HDL-C at 6 months. The total HDL-C and HDL(2)-C reductions narrow the baseline differences between men and women by 50%. Whether this reduction impacts women's protection from cardiovascular disease deserves future study. Nonetheless, the results point to sex-based differences in intrahepatic glucose and fatty acid metabolism linked to alterations in HDL formation and removal. PMID- 10845876 TI - High levels of nonesterified fatty acids are associated with increased familial risk of cardiovascular disease. AB - To address the question of whether elevated concentrations of nonesterified fatty acids (NEFAs) are associated with an increased risk of cardiovascular disease, we measured NEFA concentrations in 140 diabetic and 343 nondiabetic unrelated Swedish subjects with a family history of type 2 diabetes and related the findings to history of cardiovascular disease in their parents. Parents of nondiabetic offspring belonging to the quartile of highest NEFA concentrations had a higher risk of myocardial infarction (35% versus 16%, P<0.01) and stroke (45% versus 16%, P<0.0005) than did parents of offspring from the lowest NEFA quartile. In a multiple logistic regression analysis, a high NEFA concentration in offspring was significantly associated with myocardial infarction and stroke in their parents. No such relationship was observed between diabetic offspring and their parents. Assuming that the same relationship between NEFA concentrations and cardiovascular disease is seen in the offspring and their parents, the findings suggest that elevated NEFA concentration is a risk factor for cardiovascular disease and could be pathogenically involved in the atherosclerotic process. PMID- 10845877 TI - Plasma concentrations of a novel, adipose-specific protein, adiponectin, in type 2 diabetic patients. AB - Adiponectin is a novel, adipose-specific protein abundantly present in the circulation, and it has antiatherogenic properties. We analyzed the plasma adiponectin concentrations in age- and body mass index (BMI)-matched nondiabetic and type 2 diabetic subjects with and without coronary artery disease (CAD). Plasma levels of adiponectin in the diabetic subjects without CAD were lower than those in nondiabetic subjects (6.6+/-0.4 versus 7.9+/-0.5 microg/mL in men, 7.6+/ 0.7 versus 11.7+/-1.0 microg/mL in women; P<0.001). The plasma adiponectin concentrations of diabetic patients with CAD were lower than those of diabetic patients without CAD (4.0+/-0.4 versus 6.6+/-0.4 microg/mL, P<0.001 in men; 6.3+/ 0.8 versus 7.6+/-0. 7 microg/mL in women). In contrast, plasma levels of leptin did not differ between diabetic patients with and without CAD. The presence of microangiopathy did not affect the plasma adiponectin levels in diabetic patients. Significant, univariate, inverse correlations were observed between adiponectin levels and fasting plasma insulin (r=-0.18, P<0.01) and glucose (r= 0.26, P<0.001) levels. In multivariate analysis, plasma insulin did not independently affect the plasma adiponectin levels. BMI, serum triglyceride concentration, and the presence of diabetes or CAD remained significantly related to plasma adiponectin concentrations. Weight reduction significantly elevated plasma adiponectin levels in the diabetic subjects as well as the nondiabetic subjects. These results suggest that the decreased plasma adiponectin concentrations in diabetes may be an indicator of macroangiopathy. PMID- 10845878 TI - Genetic variation in human stromelysin gene promoter and common carotid geometry in healthy male subjects. AB - A common variant in the promoter of the human stromelysin gene, causing reduced enzyme expression, has been associated with the progression of coronary atherosclerosis. On the other hand, increased stromelysin activity may promote plaque rupture. The present study was undertaken to investigate the relationship between the genetic variation in the human stromelysin gene promoter and common carotid geometry. Forty-two healthy male subjects without major coronary heart disease risk factors were investigated. The polymorphism in the stromelysin gene promoter was studied through polymerase chain reaction amplification with the use of mutagenic primers. Age, blood pressure, lipids, glucose, viscosity, and body mass index were similar in homozygotes for the 5A allele (5A/5A), heterozygotes (5A/6A), and homozygotes for the 6A allele (6A/6A). Serum matrix metalloproteinase-3 levels did not differ significantly among genotypes. Common carotid diameters and intima-media thickness, measured by noninvasive ultrasonography, were significantly larger in 6A/6A subjects (for respective 6A/6A, 5A/6A, and 5A/5A subjects, diameter at the R wave was 0.63+/-0.09, 0.55+/ 0.06, and 0.53+/-0.04 cm [mean+/-SD], P<0.005 by ANOVA; intima-media thickness was 765+/-116, 670+/-116, and 630+/-92 microm [mean+/-SD], P<0.05 by ANOVA). Wall shear stress, calculated as blood velocityxblood viscosity/internal diameter, was significantly lower in 6A/6A subjects (for respective 6A/6A, 5A/6A, and 5A/5A subjects, mean wall shear stress was 10.4+/-2.9, 13.5+/-3.5, and 12.6+/-1.9 dyne/cm(2) [mean+/-SD], P<0.05 by ANOVA). The results demonstrate that the gene polymorphism in the promoter region of stromelysin is associated with structural and functional characteristics of the common carotid artery in healthy male subjects without major risk factors for atherosclerosis. Individuals with the 6A/6A genotype (associated with lower enzyme activity) show a triad of events, namely, increased wall thickness, enlarged arterial lumen, and local reduction of wall shear stress, which might predispose them to atherosclerotic plaque localization. PMID- 10845879 TI - Cardiovascular effects of droloxifene, a new selective estrogen receptor modulator, in healthy postmenopausal women. AB - Selective estrogen receptor modulators, like tamoxifen and related compounds, have mixed estrogen agonistic/antagonistic effects. Tamoxifen may confer significant cardiovascular benefits without the estrogen-associated risks of endometrial and breast cancer. Droloxifene, a structural analogue of tamoxifen, has estrogen agonistic effects on bone and antagonistic effects on endometrial and breast tissue. Its cardiovascular effects in women are unknown. We enrolled 24 healthy postmenopausal women in a randomized, double-blind, 2-period crossover trial comparing the effects of droloxifene (60 mg/d) with conjugated estrogen (0.625 mg/d). Plasma lipids, coagulation and fibrinolytic factors, and brachial flow-mediated vasodilator responses were measured at the beginning and end of each treatment period. Droloxifene and estrogen resulted in 16.6% and 12.0% reductions, respectively, in low density lipoprotein cholesterol (P<0.001) and 13.2% and 9.5% reductions, respectively, in lipoprotein(a) (P<0.05). In contrast, estrogen, but not droloxifene, increased high density lipoprotein (18.5%, P<0.001). Droloxifene also reduced fibrinogen by 17.8% versus a 7.3% reduction with estrogen (P=0.004) but produced no estrogen-like changes in plasminogen, plasminogen activator inhibitor-1, or tissue plasminogen activator. Droloxifene and estrogen produced 36.4% and 27.3% increases, respectively, in flow-mediated vasodilation (percent change from baseline, P<0.05 for both). Droloxifene has estrogen agonistic properties regarding low density lipoprotein and lipoprotein(a) metabolism, certain coagulation factors, and endothelium-dependent vasodilation but, unlike estrogen, has no effect on high density lipoprotein/triglyceride metabolism and the fibrinolytic cascade. It remains unknown whether droloxifene can confer a true cardiovascular benefit. PMID- 10845880 TI - Physiological concentration of 17beta-estradiol retards the progression of severe atherosclerosis induced by a high-cholesterol diet plus balloon catheter injury: role of NO. AB - The molecular mechanisms of the antiatherosclerotic effects of estrogen are not yet known. We evaluated the effects of 17beta-estradiol (E(2)) on high cholesterol diet- (HCD; standard diet and 1% cholesterol) and balloon injury induced atherosclerosis in female New Zealand White rabbits. The abdominal aortas of 40 oophorectomized (Groups 1 through 5) and 8 nonoophorectomized (Group 6) rabbits were injured by balloon catheter, and the animals were then divided into the following groups and treated for 10 weeks: Group 1, standard diet; Group 2, standard diet plus a moderate dose of E(2) (100 microg x kg(-1) x d(-1)); Group 3, HCD; Group 4, HCD plus a moderate dose of E(2); Group 5, HCD plus a low dose of E(2) (20 microg x kg(-1) x d(-1)); and Group 6, HCD in nonoophorectomized rabbits. After the treatment phase, plasma E(2) was increased up to 282.2+/-45.5 pg/mL in Group 2, 263.0+/-41.5 pg/mL in Group 4, 87. 9+/-18.8 pg/mL in Group 5, and 45.6+/-7.3 pg/mL in Group 6. HCD-mediated increases in plasma lipid levels were not changed by E(2) treatment, whereas E(2) decreased the aortic intimal thickening in Group 2 animals compared with those in Group 1 and reduced atherosclerosis in the thoracic and abdominal aortas of Group 4, 5, and 6 rabbits compared with those in Group 3. E(2) restored the impaired abdominal aortic endothelium-dependent relaxation of balloon-injured and HCD-supplemented rabbits, and E(2) increased basal nitric oxide (NO) release. The basal NO-releasing effect showed a significant, inverse relation with the severity of atherosclerosis. Plasma E(2) concentration also showed a significant, inverse relation with atherosclerotic area. In conclusion, physiological concentrations of E(2) can retard the progression of severe atherosclerosis and stabilize atheromas induced by HCD and balloon injury. The retardation may be partially mediated by endothelial NO function in vessels treated with E(2). PMID- 10845881 TI - Common carotid intima-media thickness predicts occurrence of carotid atherosclerotic plaques: longitudinal results from the Aging Vascular Study (EVA) study. AB - The role of the increase in the common carotid artery (CCA) intima-media wall thickness (IMT) in the atherosclerotic process is questionable. This longitudinal study examined the predictive value of CCA-IMT measured at baseline examination (at sites free of plaques) on the occurrence of atherosclerotic plaques in the extracranial carotid arteries during 4 years of follow-up study in a sample of 1010 subjects aged 59 to 71 years. Ultrasound examinations were performed at baseline and 2 years and 4 years later. The occurrence of carotid plaques during follow-up was defined as the appearance of >/=1 plaque in previously normal carotid segments and/or the appearance of new plaques in the carotid segments that previously had plaques. Carotid plaque occurrence was observed in 185 subjects (18.3%). Age- and sex- adjusted odds ratios of carotid plaque occurrence were 2.66 (95% CI 1.58 to 4.46, P<0.001) in subjects having intermediate baseline CCA-IMT values (quartiles 2 and 3) and 3.67 (CI 2.09 to 6.44, P<0.001) in those having the highest baseline CCA-IMT values (quartile 4) compared with those having the lowest baseline CCA-IMT values (quartile 1). Multivariate adjustment for major cardiovascular risk factors did not alter the results. These findings were observed for men and women as well as for subjects with and without carotid plaques at baseline. This 4-year longitudinal study shows that CCA-IMT predicts carotid plaque occurrence in a large sample of relatively old subjects. It extends the findings obtained from cross-sectional studies and suggests that increased intima-media thickness might occur in an earlier phase of the atherosclerotic process. PMID- 10845882 TI - Quantification in situ of crystalline cholesterol and calcium phosphate hydroxyapatite in human atherosclerotic plaques by solid-state magic angle spinning NMR. AB - Because of renewed interest in the progression, stabilization, and regression of atherosclerotic plaques, it has become important to develop methods for characterizing structural features of plaques in situ and noninvasively. We present a nondestructive method for ex vivo quantification of 2 solid-phase components of plaques: crystalline cholesterol and calcium phosphate salts. Magic angle spinning (MAS) nuclear magnetic resonance (NMR) spectra of human carotid endarterectomy plaques revealed (13)C resonances of crystalline cholesterol monohydrate and a (31)P resonance of calcium phosphate hydroxyapatite (CPH). The spectra were obtained under conditions in which there was little or no interference from other chemical components and were suitable for quantification in situ of the crystalline cholesterol and CPH. Carotid atherosclerotic plaques showed a wide variation in their crystalline cholesterol content. The calculated molar ratio of liquid-crystalline cholesterol to phospholipid ranged from 1.1 to 1.7, demonstrating different capabilities of the phospholipids to reduce crystallization of cholesterol. The spectral properties of the phosphate groups in CPH in carotid plaques were identical to those of CPH in bone. (31)P MAS NMR is a simple, rapid method for quantification of calcium phosphate salts in tissue without extraction and time-consuming chemical analysis. Crystalline phases in intact atherosclerotic plaques (ex vivo) can be quantified accurately by solid state (13)C and (31)P MAS NMR spectroscopy. PMID- 10845883 TI - Hyperbaric oxygen reduces the progression and accelerates the regression of atherosclerosis in rabbits. AB - We studied the effect of hyperbaric oxygen (HBO) treatment on the extent of diet induced accumulation of lipid oxidation products in rabbit plasma and tissues, on plasma paraoxonase activity, and on the extent of progression and regression of atherosclerotic lesions in the rabbit aorta. HBO treatment of cholesterol-fed rabbits dramatically reduces the development of arterial lesions despite having little or no effect on plasma or individual lipoprotein cholesterol concentrations. Compared with no treatment in cholesterol-fed animals, HBO treatment also substantially reduces the accumulation of lipid oxidation products (conjugated dienes, trienes, and thiobarbituric acid-reactive substances) in plasma, in the low density lipoprotein and high density lipoprotein fractions of plasma, in the liver, and in the aortic tissues. In addition, HBO treatment prevents the decrease in plasma paraoxonase activity observed in rabbits fed cholesterol-rich diets. Similarly, in regression studies, HBO treatment has no effect on the rate of plasma (or lipoprotein) cholesterol decline but significantly accelerates aortic lesion regression compared with no treatment. Direct measures of aortic cholesterol content support these morphological observations. On the basis of these results, we conclude that repeated, but relatively short, exposure to HBO induces an antioxidant defense mechanism(s) that is responsible for retarding the development or accelerating the regression of atherosclerotic lesions. PMID- 10845884 TI - Are retinal arteriolar abnormalities related to atherosclerosis?: The Atherosclerosis Risk in Communities Study. AB - The objective of this study was to describe associations of retinal arteriolar abnormalities with clinical and subclinical manifestations of atherosclerosis and a broad group of risk factors for vascular disease. A biracial population of 8772 adults (aged 48 to 72 years) living in 4 communities was examined from 1993 to 1995 were studied for that purpose. Retinal arteriovenous nicking and focal arteriolar narrowing were determined by light-box grading of a 45 degrees fundus photograph by use of a standardized protocol. Diameters of arterioles and venules were measured in digitized photographs, and a summary arteriolar-to-venular ratio was derived as an index of generalized arteriolar narrowing. Focal arteriolar narrowing was associated only with hypertension. Generalized arteriolar narrowing was associated with carotid plaque but not with any other evidence of atherosclerosis, either clinical (cardiovascular disease or stroke) or subclinical (carotid or popliteal artery thickness or lower limb obstructive disease), or with plasma cholesterol. It was also associated with smoking, with inflammatory markers (white blood cell count, fibrinogen, and reduced albumin), and with the triglyceride and high density lipoprotein cholesterol changes associated with inflammation. Arteriovenous nicking was inconsistently associated with subclinical atherosclerosis. It was not associated with cardiovascular disease, stroke, or plasma cholesterol. Arteriovenous nicking was associated with markers of inflammation and vascular endothelial dysfunction (von Willebrand factor and factor VIII). Arteriolar narrowing and nicking appear to be related to hypertension and inflammatory factors. Nicking may also be related to endothelial dysfunction. Results suggest that these arteriolar changes are pathologically distinct from atherosclerosis. Including their measurement in population studies may permit evaluation of the independent contribution of arteriolar disease to various ischemic diseases of the heart, brain, and other organs. PMID- 10845885 TI - Inhibition of platelet integrin alpha(IIb)beta(3) by peptides that interfere with protein kinases and the beta(3) tail. AB - alpha-Thrombin stimulation of human platelets initiates inside-out signaling to integrin alpha(IIb)beta(3) (glycoprotein IIb/IIIa), resulting in the exposure of ligand binding sites. In the present study, the regulation of alpha(IIb)beta(3) via protein kinases was investigated in platelets permeabilized with streptolysin O by introducing peptides that interfere with these enzymes and with possible regulatory domains in the cytosolic tail of the beta(3) subunit. Compared with intact platelets, the permeabilized platelets preserved >80% of the aggregation, secretion, and alpha(IIb)beta(3) ligand binding capacity. The peptide YIYGSFK, a substrate for Src kinases, inhibited alpha-thrombin-induced ligand binding to alpha(IIb)beta(3), but a reversed peptide with Y-->F substitutions (KFSGFIF) had no effect. Ligand binding to alpha(IIb)beta(3) was also inhibited by the peptide RKRCLRRL, which binds irreversibly to the catalytic domain of protein kinase C. Peptides corresponding to parts of the protein C inhibitor and beta(2) glycoprotein I were used as negative controls and failed to interfere with ligand binding. Possible target domains for protein kinases are present in the cytoplasmic tail of the beta(3) subunit. The LLITIHDR peptide, matching the membrane-proximal domain of beta(3) (residues 717 to 724), had no effect, but NNPLYKEA (residues 743 to 750), EATSTFTN (residues 749 to 756), and TNITYRGT (residues 755 to 762), which mimicked overlapping domains of the carboxy-terminal part of beta(3), reduced alpha-thrombin-induced ligand binding by 60+/-4%, 97+/ 1%, and 97+/-2% (n=3) at 500 micromol/L peptide, respectively. These observations indicate that Src kinases and protein kinase C take part in inside-out signaling to integrin alpha(IIb)beta(3) and identify target domains in beta(3) that contribute to the regulation of this integrin. PMID- 10845886 TI - Platelet thrombus formation on collagen at high shear rates is mediated by von Willebrand factor-glycoprotein Ib interaction and inhibited by von Willebrand factor-glycoprotein IIb/IIIa interaction. AB - We studied the role of von Willebrand Factor (vWF) in platelet thrombus formation in flowing blood by using a perfusion system and mutant forms of vWF lacking either interaction with glycoprotein Ib (GpIb) or with glycoprotein IIb/IIIa (alphaIIb-beta3). These mutants were added to the blood of patients with severe von Willebrand's disease (vWD) or to normal blood reconstituted with a human albumin solution instead of plasma. This blood was then perfused over collagen type III spray-coated on a glass surface and preincubated for 2 hours with 20 microg/mL plasma vWF. In this way, the adhesion step was mediated by the preincubated plasma vWF bound to collagen type III, whereas thrombus formation was mediated by mutant vWF added to the perfusate. Thrombus formation was absent at all 3 shear rates studied (300, 800, and 2600 s(-1)) when DeltaA1-vWF, lacking interaction with GpIb, was added to the perfusate, indicating the importance of GpIb-vWF interaction for thrombus formation. The interaction of vWF and GpIb is currently thought to be possible under physiological conditions in which the conformation of vWF has been changed by adsorption to a surface. Our results regarding the role of GpIb-vWF interaction in thrombus formation suggest that a second mechanism may operate by which a change may occur in GpIb on the surface of adhered platelets either by activation of the molecule or as a consequence of shear stress. Increased thrombus formation was observed when the Arg-Gly-Gly-Ser vWF, which does not interact with alphaIIb-beta3, was added to vWD blood and perfused at 2600 s(-1). This increase was not observed in vWD blood at lower shear rates or after addition of Arg-Gly-Gly-Ser-vWF to reconstituted normal blood. Thrombus formation at a high shear rate was largest when either vWF or fibrinogen was present as a single ligand for alphaIIb-beta3 at a high shear rate. When both were present, thrombus formation was decreased. We postulate that thrombus formation is less efficient because of incomplete bridge formation when vWF and fibrinogen are both present as ligands for alphaIIb-beta3. PMID- 10845887 TI - Progressive and transient expression of tissue plasminogen activator during fetal development. AB - In previous studies of the role of tissue plasminogen activator (tPA) in the lung inflammatory response, we observed that tPA expression was present exclusively in the small arteries and arterioles within the lung and absent from the capillaries, veins, and large pulmonary arteries. To define more completely the expression pattern of tPA, we evaluated the distribution of this protein during prenatal and postnatal development. tPA was first observed in the rat fetus at day 13 in the large arteries of both the thoracic and cranial cavities, including the dorsal aortas and pulmonary arteries in the former and the internal carotid and middle cerebral arteries in the latter. By day 15, tPA was no longer detectable in the aortas but appeared throughout the pulmonary, subclavian, vertebral, and basilar arteries. At day 17, tPA had disappeared from the subclavian artery and the proximal portion of the vertebral artery but was found in the smaller arterial branches of these 2 large vessels. By the end of gestation, tPA had also disappeared from the main pulmonary arteries but remained in the branches at the hilus of the lung. At birth, tPA was concentrated in the endothelia of arteries within the pia mater, the basilar and superficial cerebral arteries, and the lung arterial system. As the animals reached maturity, tPA disappeared from the larger cerebral arteries and their cortical branches but continued to be expressed in the vessels of the pia mater and lung. This study indicates that tPA expression is a dynamic process that responds to a changing arterial environment during vascular development. PMID- 10845888 TI - Hypertriglyceridemic VLDL downregulates tissue plasminogen activator gene transcription through cis-repressive region(s) in the tissue plasminogen activator promoter in cultured human endothelial cells. AB - The relationship between tissue plasminogen activator (tPA) levels and the potential regulation by hypertriglyceridemic very low density lipoprotein (HTG VLDL) was examined in a human umbilical vein endothelial cell (HUVEC) culture model system. HUVEC cultures were incubated in the absence/presence of HTG-VLDL or normal (NTG)-VLDL (0 to 50 microg/mL) at 37 degrees C for various times (0 to 24 hours), followed by analyses of tPA antigen (ELISA), mRNA (reverse transcription-polymerase chain reaction), endothelial cell surface-localized plasmin generation assays, and nuclear transcription run-on assays. Secreted tPA antigen levels decreased approximately 53% (3.3+/-0.14 versus 6.97+/-0.42 microg/mL) and mRNA levels decreased approximately 70% in HTG-VLDL-treated HUVECs compared with NTG-VLDL-treated and culture medium control cells. Decreased tPA antigen and mRNA expression was associated with a concomitant approximately 98% decrease in tPA-mediated plasmin generation in HTG-VLDL-treated HUVEC cultures. Nuclear transcription run-on assays demonstrated that HTG-VLDL decreased tPA gene transcription approximately 73% (tPA mRNA/GAPDH mRNA) in cultured HUVECs. To identify and localize the repressive element(s) in the tPA promoter responsive to HTG-VLDL, a tPA promoter/luciferase construct (ptPA222/luc) was generated. HUVECs transiently transfected with this construct were incubated in the absence/presence of HTG-VLDL or NTG-VLDL (20 microg/mL). HTG-VLDL decreased promoter activity approximately 52% to 57% in the ptPA222/luc-transfected cells compared with NTG-VLDL-treated or buffer control cells. These results indicate that the 2.2-kb fragment of the promoter and 5' flanking region of the tPA gene contains the repressive sequences that direct the transcriptional downregulation of the tPA promoter. Data from these studies suggest that the repression of tPA gene expression by HTG-VLDL may contribute to the impaired fibrinolysis often associated with hypertriglyceridemia. PMID- 10845889 TI - Role of cytokines in the regulation of plasminogen activator inhibitor-1 expression and secretion in newly differentiated subcutaneous human adipocytes. AB - Elevated levels of plasminogen activator inhibitor-1 (PAI-1) are characteristic for obesity and are associated with increased risk of thromboembolic complications. PAI-1 recently was reported to be expressed and secreted by human adipocytes, but little is known about regulation of PAI-1 in human adipose tissue. Therefore, we examined the effects of selected cytokines present in adipose tissue on expression and secretion of PAI-1 in in vitro, differentiated subcutaneous human adipocytes in primary culture. Transforming growth factor beta1 (TGF-beta1) increased PAI-1 secretion in a dose- and time-dependent manner. PAI-1 protein increased by 3.2-fold and PAI-1 mRNA by 1.9-fold after a 6-hour exposure to 400 pmol/L TGF-beta1. This effect is probably mediated by TGF-beta1 type 2 and 3 receptors, which were found to be expressed in cultured human adipocytes. Moreover, TNF-alpha and interkeukin-1beta (IL-1beta) also exerted a stimulatory effect on PAI-1 release and increased PAI-1 mRNA levels. As assessed by a semiquantitative reverse transcription-polymerase chain reaction technique, TGF-beta1 mRNA is expressed by differentiation of human preadipocytes and is moderately upregulated by TNF-alpha and IL-1beta. In conclusion, our results clearly indicate that TGF-beta1 is a potent inducer of PAI-1 production in subcutaneous human adipocytes. In addition, data suggest that TNF-alpha and IL 1beta also have stimulatory effects on PAI-1 protein secretion and may contribute to the elevated PAI-1 levels observed in obesity. PMID- 10845890 TI - Impact of antioxidants and HDL on glycated LDL-induced generation of fibrinolytic regulators from vascular endothelial cells. AB - Hyperglycemia and dyslipoproteinemia are biochemical markers of diabetes mellitus (DM). Elevated levels of plasminogen activator inhibitor-1 (PAI-1) with and without reduction of tissue plasminogen activator (tPA) in plasma have been frequently found in patients with DM. Our previous studies indicated that glycation enhances low density lipoprotein (LDL)-induced production of PAI-1 and further decreases tPA generation in vascular endothelial cells (ECs). The present study demonstrated that treatment with antioxidants, butylated hydroxytoluene or vitamin E, blocked native LDL- and glycated LDL-induced changes in PAI-1 and tPA generation in ECs. Native or glycated high density lipoprotein (HDL) did not significantly alter tPA generation in ECs. Glycated but not native HDL (>/=100 microg/mL) moderately increased PAI-1 release from ECs. Cotreatment with native or glycated HDL inhibited LDL-induced or glycated LDL-induced changes in PAI-1 and tPA generation in ECs. The abundance of conjugated dienes was increased in glycated or EC-modified LDL. Treatment with butylated hydroxytoluene, vitamin E, or HDL reduced the abundance of conjugated dienes in glycated or EC-modified LDL. The effects of antioxidants and HDL on LDL-induced or its glycated LDL-induced changes in the generation of PAI-1 and tPA were also found in cultured human coronary artery ECs. The findings of the present study suggest that antioxidants and HDL may attenuate native LDL- or glycated LDL-induced changes in the generation of fibrinolytic regulators from vascular ECs, which possibly results from their inhibition on the lipid peroxidation of LDL particles. Treatment with antioxidants or hypolipidemic agents potentially improves fibrinolytic activity and reduces thrombotic tendencies in patients with DM. PMID- 10845891 TI - When is atherosclerosis not atherosclerosis? PMID- 10845892 TI - Megakaryocyte-targeted synthesis of the integrin beta(3)-subunit results in the phenotypic correction of Glanzmann thrombasthenia. AB - Glanzmann thrombasthenia is an inherited bleeding disorder characterized by qualitative or quantitative defects of the platelet-specific integrin, alphaIIbbeta(3). As a result, alphaIIbbeta(3) cannot be activated and cannot bind to fibrinogen, leading to a loss of platelet aggregation. Thrombasthenia is clinically characterized by mucocutaneous hemorrhage with episodes of intracranial and gastrointestinal bleeding. To develop methods for gene therapy of Glanzmann thrombasthenia, a murine leukemia virus (MuLV)-derived vector, 889Pl(A2)beta(3), was transduced into peripheral blood CD34(+) cells from 2 patients with thrombasthenia with defects in the beta(3) gene. The human alphaIIb promoter was used in this vector to drive megakaryocyte-targeted expression of the wild-type beta(3) subunit. Proviral DNA and alphaIIbbeta(3) biosynthesis were detected after in vitro differentiation of transduced thrombasthenic CD34(+) cells with megakaryocyte growth and development factor. Flow cytometric analysis of transduced patient samples indicated that 19% of megakaryocyte progeny expressed alphaIIbbeta(3) on the surface at 34% of normal receptor levels. Treatment of transduced megakaryocytes with a combination of agonists including epinephrine and the thrombin receptor-activating peptide induced the alphaIIbbeta(3) complex to form an activated conformation capable of binding fibrinogen as measured by PAC-1 antibody binding. Transduced cells retracted a fibrin clot in vitro similar to megakaryocytes derived from a normal nonthrombasthenic individual. These results demonstrate ex vivo phenotypic correction of Glanzmann thrombasthenia and support the potential use of hematopoietic CD34(+) cells as targets for alphaIIb promoter-driven MuLV vectors for gene therapy of platelet disorders. (Blood. 2000;95:3645-3651) PMID- 10845893 TI - Primary and secondary cutaneous CD30(+) lymphoproliferative disorders: a report from the Dutch Cutaneous Lymphoma Group on the long-term follow-up data of 219 patients and guidelines for diagnosis and treatment. AB - To evaluate our diagnostic and therapeutic guidelines, clinical and long-term follow-up data of 219 patients with primary or secondary cutaneous CD30(+) lymphoproliferative disorders were evaluated. The study group included 118 patients with lymphomatoid papulosis (LyP; group 1), 79 patients with primary cutaneous CD30(+) large T-cell lymphoma (LTCL; group 2), 11 patients with CD30(+) LTCL and skin and regional lymph node involvement (group 3), and 11 patients with secondary cutaneous CD30(+) LTCL (group 4). Patients with LyP often did not receive any specific treatment, whereas most patients with primary cutaneous CD30(+) LTCL were treated with radiotherapy or excision. All patients with skin limited disease from groups 1 and 2 who were treated with multiagent chemotherapy had 1 or more skin relapses. The calculated risk for systemic disease within 10 years of diagnosis was 4% for group 1, 16% for group 2, and 20% for group 3 (after initial therapy). Disease-related 5-year-survival rates were 100% (group 1), 96% (group 2), 91% (group 3), and 24% (group 4), respectively. The results confirm the favorable prognoses of these primary cutaneous CD30(+) lymphoproliferative disorders and underscore that LyP and primary cutaneous CD30(+) lymphomas are closely related conditions. They also indicate that CD30(+) LTCL on the skin and in 1 draining lymph node station has a good prognosis similar to that for primary cutaneous CD30(+) LTCL without concurrent lymph node involvement. Multiagent chemotherapy is only indicated for patients with full blown or developing extracutaneous disease; it is never or rarely indicated for patients with skin-limited CD30(+) lymphomas. (Blood. 2000;95:3653-3661) PMID- 10845894 TI - RHD gene deletion occurred in the Rhesus box. AB - The Rh blood group antigens derive from 2 genes, RHD and RHCE, that are located at chromosomal position 1p34.1-1p36 (chromosome 1, short arm, region 3, band 4, subband 1, through band 6). In whites, a cde haplotype with a deletion of the whole RHD gene occurs with a frequency of approximately 40%. The relative position of the 2 RH genes and the location of the RHD deletion was previously unknown. A model has been developed for the RH locus using RHD- and RHCE-related nucleotide sequences deposited in nucleotide sequence databases along with polymerase chain reaction (PCR) and nucleotide sequencing. The open reading frames of both RH genes had opposite orientations. The 3' ends of the genes faced each other and were separated by about 30 000 base pair (bp) that contained the SMP1 gene. The RHD gene was flanked by 2 DNA segments, dubbed Rhesus boxes, with a length of approximately 9000 bp, 98.6% homology, and identical orientation. The Rhesus box contained the RHD deletion occurring within a stretch of 1463 bp of identity. PCR with sequence-specific priming (PCR-SSP) and PCR with restriction fragment length polymorphism (PCR-RFLP) were used for specific detection of the RHD deletion. The molecular structure of the RH gene locus explains the mechanisms for generating RHD/RHCE hybrid alleles and the RHD deletion. Specific detection of the RHD(-) genotype is now possible. (Blood. 2000;95:3662-3668) PMID- 10845895 TI - Vena caval filters: a comprehensive review. AB - Hematologists are often asked to treat patients with venous thromboembolic disease. Although anticoagulation remains the primary therapy for venous thromboembolism, vena caval filters are an important alternative when anticoagulants are contraindicated. To assess the evidence supporting the utility of these devices, a comprehensive review of the English language literature was performed. Except for one randomized trial, the vena caval filter literature consists of case series or consecutive case series. The mean duration of follow up for each of the 5 filter types varies from 6 to 18 months. All are about equally effective in the prevention of pulmonary embolism (2.6%-3.8%). Deep venous thrombosis (6%-32%) and inferior vena cava thrombosis (3.6%-11.2%) after filter placement vary widely among different filter types primarily because of differences in outcome assessment. Thrombosis at the insertion site is a common complication of filter placement (23%-36%). In view of the absence of randomized comparisons, no filter can be designated as superior in safety or efficacy. Vena caval filters represent a potentially important but poorly evaluated therapeutic modality in the prevention of pulmonary emboli. Randomized trials are necessary to establish the appropriate place for vena caval filters in the treatment of venous thromboembolic disease. (Blood. 2000;95:3669-3677) PMID- 10845896 TI - High levels of factor IX increase the risk of venous thrombosis. AB - Elevated plasma levels of factor VIII (> 150 IU/dL) are an important risk factor for deep vein thrombosis (DVT). Factor VIII is the cofactor of factor IXa in the activation of factor X. The risk of thrombosis in individuals with an elevated factor IX level is unknown. This study investigated the role of elevated factor IX levels in the development of DVT. We compared 426 patients with a first objectively diagnosed episode of DVT with 473 population controls. This study was part of a large population-based case-control study on risk factors for venous thrombosis, the Leiden Thrombophilia Study (LETS). Using the 90th percentile measured in control subjects (P(90) = 129 U/dL) as a cutoff point for factor IX levels, we found a 2- to 3-fold increased risk for individuals who have factor IX levels above 129 U/dL compared with individuals having factor IX levels below this cutoff point. This risk was not affected by adjustment for possible confounders (age, sex, oral contraceptive use, and high levels of factor VIII, XI, and vitamin K-dependent proteins). After exclusion of individuals with known genetic disorders, we still found an odds ratio (OR) of 2.5 (95% confidence interval [CI]: 1.6-3.9). The risk was higher in women (OR: 2.6, CI: 1.6-4.3) than in men (OR: 1.9, CI: 1.0-3.6) and appeared highest in the group of premenopausal women not using oral contraceptives (OR: 12.4, CI: 3.3-47.2). These results show that an elevated level of factor IX is a common risk factor for DVT. (Blood. 2000;95:3678-3682) PMID- 10845897 TI - Chronic graft versus host disease is associated with long-term risk for pneumococcal infections in recipients of bone marrow transplants. AB - Incidences of and risk factors for Streptococcus pneumoniae sepsis (SPS) after hematopoietic stem cell transplantation were analyzed in 1329 patients treated at a single center between 1973 and 1997. SPS developed in 31 patients a median of 10 months after transplantation (range, 3 to 187 months). The infection was fatal in 7 patients. The probability of SPS developing at 5 and 10 years was 4% and 6%, respectively. Age, sex, diagnosis, and graft versus host disease (GVHD) prophylaxis did not influence the development of SPS. Allogeneic transplantation (10-year probability, 7% vs 3% for nonallogeneic transplants; P =.03) and chronic GVHD (10-year probability, 14% vs 4%; P =.002) were associated with significantly higher risk for SPS. All the episodes of SPS were seen in patients who had undergone allograft or total body irradiation (TBI) (31 of 1202 vs 0 of 127; P =.07). Eight patients were taking regular penicillin prophylaxis at the time of SPS, whereas 23 were not taking any prophylaxis. None of the 7 patients with fatal infections was taking prophylaxis for Pneumococcus. Pneumococcal bacteremia was associated with higher incidences of mortality (6 of 15 vs 1 of 16; P =.04). We conclude that there is a significant long-term risk for pneumococcal infection in patients who have undergone allograft transplantation, especially those with chronic GVHD. Patients who have undergone autograft transplantation after TBI containing regimens also appear to be at increased risk. These patients should receive lifelong pneumococcus prophylaxis. Consistent with increasing resistance to penicillin, penicillin prophylaxis does not universally prevent SPS, though it may protect against fatal infections. Further studies are required to determine the optimum prophylactic strategy in patients at risk. (Blood. 2000;95:3683-3686) PMID- 10845898 TI - Quality of life-adjusted survival analysis of high-dose therapy with autologous bone marrow transplantation versus sequential chemotherapy for patients with aggressive lymphoma in first complete remission. Groupe d'Etude les Lymphomes de l'Adulte (GELA). AB - Evaluating high-dose therapy (HDT) with autologous stem cell transplantation (ASCT) in term of both duration and quality of life (QOL) presents major interests for patients with non-Hodgkin lymphoma. The quality-adjusted time without symptom and toxicity (Q-TWiST) methodology was applied to the LNH87-2 trial comparing HDT with ASCT versus sequential chemotherapy in 541 patients in first complete remission (CR). Overall survival (OS) and disease-free survival (DFS) curves were used to estimate duration of 4 health states: acute short-term toxicity (Tox1), secondary toxicity (Tox2), time without symptom and toxicity (TWiST), and relapse (Rel). Areas under survival curves (AUC) were retrospectively weighted according to QOL coefficients. HDT increased, but not significantly, TWiST (+2. 4 months in AUC, P =.17) and decreased Rel (-3 months, P <.01). Survival estimates did not differ between the 2 treatments (AUC 47.7 months for OS, 39.7 months for DFS). High-risk patients treated by HDT versus chemotherapy had a significant benefit in DFS (AUC 28.8 versus 24.9 months, P <.01) but not in OS (AUC 37.3 versus 36 months, P =.27). Sensitivity analysis, performed by varying QOL coefficients, demonstrated significant quality-adjusted survival gain in high-risk patients treated by HDT. In low-risk patients, a diagram provided an aid to clinical decision-making. This analysis supports the use of HDT in these patients with adverse prognostic factors in the first CR, even after adjusting for QOL using the Q-TWiST method. (Blood. 2000;95:3687-3692) PMID- 10845899 TI - A combination of anti-CD3 and anti-CD7 ricin A-immunotoxins for the in vivo treatment of acute graft versus host disease. AB - This study evaluated the anti-graft versus host disease (GVHD) potential of a combination of immunotoxins (IT), consisting of a murine CD3 (SPV-T3a) and CD7 (WT1) monoclonal antibody both conjugated to deglycosylated ricin A. In vitro efficacy data demonstrated that these IT act synergistically, resulting in an approximately 99% elimination of activated T cells at 10(-8 )mol/L (about 1.8 microg/mL). Because most natural killer (NK) cells are CD7(+), NK activity was inhibited as well. Apart from the killing mediated by ricin A, binding of SPV-T3a by itself impaired in vitro cytotoxic T-cell cytotoxicity. Flow cytometric analysis revealed that this was due to both modulation of the CD3/T-cell receptor complex and activation-induced cell death. These results warranted evaluation of the IT combination in patients with refractory acute GVHD in an ongoing pilot study. So far, 4 patients have been treated with 3 to 4 infusions of 2 or 4 mg/m(2) IT combination, administered intravenously at 48-hour intervals. The T(1/2) was 6.7 hours, and peak serum levels ranged from 258 to 3210 ng/mL. Drug associated side effects were restricted to limited edema, fever, and a modest rise of creatine kinase levels. One patient developed low-titer antibodies against ricin A. Infusions were associated with an immediate drop of circulating T cells, followed by a more gradual but continuing elimination of T/NK cells. One patient mounted an extensive CD8 T-cell response directly after treatment, not accompanied with aggravating GVHD. Two patients showed nearly complete remission of GVHD, despite unresponsiveness to the extensive pretreatment. These findings justify further investigation of the IT combination for treatment of diseases mediated by T cells. (Blood. 2000;95:3693-3701) PMID- 10845900 TI - Blood stem cells compared with bone marrow as a source of hematopoietic cells for allogeneic transplantation. IBMTR Histocompatibility and Stem Cell Sources Working Committee and the European Group for Blood and Marrow Transplantation (EBMT). AB - Peripheral blood cells are increasingly used in place of bone marrow as a source of hematopoietic stem cells for allogeneic transplantation. The relative efficacy of these 2 approaches is unknown. This retrospective multivariate analysis compared results of 288 HLA-identical sibling blood stem cell transplantations with results of 536 HLA-identical sibling bone marrow transplantations. No transplants were T-cell depleted. Median follow-up was 12 months, and analyses focused on 1-year outcomes. Recipients of blood stem cell transplants had more rapid recovery of neutrophils to at least 0.5 x 10(9)/L (median time to recovery, 14 days, compared with 19 days for marrow transplants; P <.001) and of platelets to at least 20 x 10(9)/L (median time, 18 days, compared with 25 days for marrow transplants; P <.001). There was no significant difference in the incidence of grades II to IV acute graft versus host disease (GVHD). The incidence of chronic GVHD was significantly higher after blood stem cell transplantation (1-year probability [95% confidence interval], 65% [56%-72%] compared with 53% [47%-59%]; P =.02) Relapse incidence in the 2 transplant groups did not differ significantly. Treatment-related mortality rates were lower and leukemia-free survival rates were higher with blood stem cell transplants in patients with advanced leukemia (acute leukemia in second remission or chronic myelogenous leukemia in accelerated phase) but not in early leukemia (acute leukemia in first remission or chronic myelogenous leukemia in chronic phase). The median time from transplantation to hospital discharge was 23 days after blood stem cell transplantation and 28 days after bone marrow transplantation (P =.003). Further study with longer follow-up is necessary to definitively establish the role of blood stem cells for allogeneic transplantation, especially in patients with good risk disease. (Blood. 2000;95:3702-3709) PMID- 10845901 TI - Hematopoietic stem cells with controllable tEpoR transgenes have a competitive advantage in bone marrow transplantation. AB - In a previous study, it was found that a truncated erythropoietin receptor transgene (tEpoR tg) enables multilineage hematopoietic progenitor amplification after treatment with erythropoietin (epo) in vitro and in vivo. This study used competitive bone marrow (BM) repopulation to show that tEpoR tg facilitates transplantation by hematopoietic stem cells (HSC). Individual multilineage colonies, committed myeloid progenitor colonies, and lymphoid colonies (pre-B colony-forming units) were grown from the marrow of animals 6 months after they received a 50/50 mixture of transgene and wild-type BM cells. In epo-treated recipients, the transgene-bearing cells significantly outcompeted the wild-type cells (84%-100% versus 16%-0%, respectively). In recipients treated with phosphate-buffered saline, the repopulation was minimally different from the donor mixture (49%-64% transgene versus 51%-36% wild-type). The epo-induced repopulation advantage is maintained in secondary transplants. In addition, neither accelerated HSC depletion nor uncontrollable proliferation occurred during epo-stimulated serial transplants of transgene-containing BM. Thus, the tEpoR tg functions in a benign fashion in HSC and allows for a significant and controllable repopulation advantage in vivo without excessive HSC depletion relative to wild-type BM. (Blood. 2000;95:3710-3715) PMID- 10845902 TI - TNF-related apoptosis-inducing ligand (TRAIL) as a negative regulator of normal human erythropoiesis. AB - The impact of tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) on normal hematopoietic development was investigated using adult peripheral blood CD34(+) hematopoietic progenitor cells, induced to differentiate along the erythroid, megakaryocytic, granulocytic, and monocytic lineages by the addition of specific cytokine cocktails. TRAIL selectively reduced the number of erythroblasts, showing intermediate levels of glycophorin A (glycophorin A(interm)) surface expression, which appeared in liquid cultures supplemented with stem cell factor + interleukin 3 + erythropoietin at days 7-10. However, neither immature (day 4) glycophorin A(dim) erythroid cells nor mature (day 14) glycophorin A(bright) erythroblasts were sensitive to TRAIL-mediated apoptosis. Moreover, pre-exposure to TRAIL significantly decreased the number and size of erythroid colonies in semisolid assays. These adverse effects of TRAIL were selective for erythropoiesis, as TRAIL did not significantly influence the survival of cells differentiating along the megakaryocytic, granulocytic, or monocytic lineages. Furthermore, TRAIL was detected by Western blot analysis in lysates obtained from normal bone marrow mononuclear cells. These findings indicate that TRAIL acts in a lineage- and stage of differentiation-specific manner, as a negative regulator of normal erythropoiesis. (Blood. 2000;95:3716 3724) PMID- 10845903 TI - "Emergency" granulopoiesis in G-CSF-deficient mice in response to Candida albicans infection. AB - Granulocyte colony-stimulating factor (G-CSF) is a glycoprotein believed to play an important role in regulating granulopoiesis both at steady state and during an "emergency" situation. Generation of G-CSF and G-CSF receptor-deficient mice by gene targeting has demonstrated unequivocally the importance of G-CSF in the regulation of baseline granulopoiesis. This study attempted to define the physiologic role of G-CSF during an emergency situation by challenging a cohort of wild-type and G-CSF-deficient mice with Candida albicans. Interestingly, after infection, G-CSF-deficient mice developed an absolute neutrophilia that was observed both in blood and bone marrow. In addition, 3 days after Candida infection increased numbers of granulocyte-macrophage (GM) and macrophage (M) progenitors were observed in the bone marrow of G-CSF-deficient mice. Of the cytokines surveyed, interleukin (IL)-6 levels in serum were elevated; interestingly, levels of IL-6 were higher and more sustained in G-CSF-deficient mice infected with C albicans than similarly infected wild-type mice. Despite the higher levels of serum IL-6, this cytokine is dispensable for the observed neutrophilia because candida-infected IL-6-deficient mice, or mice simultaneously deficient in G-CSF and IL-6, developed neutrophilia. Similarly, mice lacking both G-CSF and GM-CSF developed absolute neutrophilia and had elevated numbers of GM and M progenitors in the bone marrow; thus, G-CSF and GM-CSF are dispensable for promoting the emergency response to candidal infection. (Blood. 2000;95:3725 3733) PMID- 10845904 TI - Sp1 and C/EBP are necessary to activate the lactoferrin gene promoter during myeloid differentiation. AB - In this study, we sought to identify factors responsible for the positive modulation of lactoferrin (LF), a neutrophil-specific, secondary-granule protein gene. Initial reporter gene transfection assays indicated that the first 89 base pairs of the LF promoter are capable of directing myeloid-specific LF gene expression. The presence of a C/EBP site flanked by 2 Sp1 sites within this segment of the LF promoter prompted us to investigate the possible role of these sites in LF expression. Cotransfection studies of LF-89luc plasmid with increasing concentrations of a C/EBPalpha expression vector in myeloid cells resulted in a linear transactivation of luciferase reporter activity. Electrophoretic mobility shift assays found that the C/EBP site is recognized by C/EBPalpha and that both LF Sp1 binding sites bind the Sp1 transcription factor specifically in myeloid cells. Mutation of either Sp1 site markedly reduced activity of the LF-89luc plasmid in myeloid cells, and neither Sp1 mutant plasmid was transactivated by a C/EBPalpha expression plasmid to the same extent as wild type LF-89luc. We also transfected LF-89luc into Drosophila Schneider cells, which do not express endogenous Sp1, and demonstrated up-regulation of luciferase activity in response to a cotransfected Sp1 expression plasmid, as well as to a C/EBPalpha expression plasmid. Furthermore, cotransfection of LF-89luc plasmid simultaneously with C/EBPalpha and Sp1 expression plasmids resulted in an increase in luciferase activity greater than that induced by either factor alone. Taken together, these observations indicate a functional interaction between C/EBP and Sp1 in mediating LF expression. (Blood. 2000;95:3734-3741) PMID- 10845905 TI - Interferon gamma delays apoptosis of mature erythroid progenitor cells in the absence of erythropoietin. AB - Based on the hypothesis that interferon gamma (IFN-gamma) may have stimulating effects on survival of hematopoietic progenitor cells, we examined the effect of IFN-gamma on apoptosis of mature erythroid colony-forming cells (ECFCs) derived from human peripheral blood obtained from normal, healthy volunteers. When the cells were cultured in the presence of IFN-gamma, even without erythropoietin (EPO), the viability of the cells was maintained for at least 36 hours. When apoptosis of ECFCs was assessed by flow cytometric analysis', using annexin V, IFN-gamma reduced the extent of apoptosis of the cells, as well as EPO. DNA fragmentation of ECFCs was also reduced by IFN-gamma. In cells cultured with IFN gamma alone, expression of Bcl-x was detected but the level of expression decreased gradually during incubation for 36 hours, and the expression level was lower than incubation with EPO. Fas expression and activation of downstream caspases were assessed by flow cytometric analysis or fluorometric protease assay. IFN-gamma induced Fas expression of the cells without the activation of caspase8 or caspase3 during 16 hours of incubation, while deprivation of EPO induced expression of Fas and the activation of both caspase8 and caspase3. We propose that IFN-gamma produces a stimulating signal for the survival of mature erythroid progenitor cells by reducing apoptosis through a mechanism other than modulating Fas and one related to the expression of Bcl-x. (Blood. 2000;95:3742 3749) PMID- 10845906 TI - Capture of cytokine-responsive genes (NACA and RBM3) using a gene trap approach. AB - We have developed a gene trap approach to select specific cytokine receptor/ligand responsive genes in the cell line TF-1. This cell line exhibits a dependency on granulocyte-macrophage colony-stimulating factor (GM-CSF) or interleukin-3 (IL-3) and responds to interleukin-5 (IL-5). In an attempt to detect genes modulated by one of these factors, cells were infected with the Rosabetageo retrovirus in the presence of GM-CSF, IL-3, or IL-5 and clones were selected for retroviral integration on the basis of G418 resistance. Housekeeping and cytokine-regulated trapped genes were then differentiated on the basis of G418 resistance versus sensitivity in the presence of the different cytokines. To determine the reliability of this screen, DNA sequences upstream of the proviral integration site were identified by 5' rapid amplification of DNA ends polymerase chain reaction (RACE PCR) from selected GM-CSF-treated and -infected clones. Comparison of the sequences with those in the Genbank database revealed that 2 sequences correspond to known genes: NACA and RBM3. NACA was recently defined as a coactivator of c-jun-mediated transcription factors in osteoblasts, and RBM3 as a protein from the heterogeneous nuclear ribonucleoprotein family. Data from transcriptional analysis of these 2 genes in TF-1 cells showed a specific up regulation by GM-CSF. Both transcripts were also found to be up-regulated in purified CD34(+) cells, suggesting their involvement in proliferative processes during hematopoiesis. Interestingly, down-regulation was observed during monocytic differentiation of TF-1 cells, suggesting their extinction could contribute to monocytic lineage development. This study demonstrates that this gene trap approach is a useful method for identifying novel, specific cytokine responsive genes that are involved in the regulation of hematopoiesis. (Blood. 2000;95:3750-3757) PMID- 10845907 TI - CD40-ligand stimulates myelopoiesis by regulating flt3-ligand and thrombopoietin production in bone marrow stromal cells. AB - CD40 ligand (CD40L)/CD40 interactions play a central role in T-cell-dependent B cell activation as previously shown by in vitro studies, the phenotype of CD40L knockout mice and the defective expression of CD40L in patients who have X-linked immunodeficiency with hyper-IgM. The distribution of CD40 in cells other than of myeloid and lymphoid lineages has suggested additional functions for this receptor/ligand couple. Here we show that CD40L stimulates myelopoiesis with a noticeable effect on megakaryocytopoiesis in cocultures of hematopoietic progenitor cells and bone marrow stromal cells. These results suggest a mechanism by which T-cell or platelet-associated or soluble CD40L may regulate myelopoiesis. (Blood. 2000;95:3758-3764) PMID- 10845908 TI - Constitutive Stat3, Tyr705, and Ser727 phosphorylation in acute myeloid leukemia cells caused by the autocrine secretion of interleukin-6. AB - To explore the activation patterns of signal transducer and activator of transcription 3 (Stat3) in acute myeloid leukemia (AML), we examined whether the phosphorylation of tyrosine705 (Tyr705) and serine727 (Ser727) residues was abnormally regulated in cells from patients with AML. In 5 of 20 (25%) patients with AML, Stat3 was constitutively phosphorylated on Tyr705 and Ser727, which were not further up-regulated by treatment with IL-6. Furthermore, Stat3 was constitutively bound to the IRE response element in these cells as determined by electrophoretic mobility shift assay, and stimulation with IL-6 did not result in increased DNA binding. Interestingly, AML cells with constitutive Stat3 activation also secreted high levels of IL-6 protein. Treating these AML cells with anti-IL-6 resulted in restored IL-6-inducible Stat3 phosphorylation on both Tyr705 and Ser727 with low or undetectable basal phosphorylation levels in unstimulated cells. In contrast, treatment with anti-IL-1 did not result in altered Stat3 phosphorylation patterns. The constitutive IL-6 expression was associated with elevated levels of suppressor of cytokine signaling-1 (SOCS-1) and SOCS-3 mRNA expression, which were not down-regulated by anti-IL-6. These data indicate that the constitutive Stat3 activation in the investigated AML blasts is caused by high IL-6 secretion levels, thus stimulating the Jak/Stat pathway in an autocrine manner, a paracrine manner, or both. (Blood. 2000;95:3765 3770) PMID- 10845909 TI - Chemoprotective effects of KF41399, a derivative of carbazole compounds, on nimustine-induced thrombocytopenia. AB - We examined the chemoprotective effects of KF41399, a novel derivative of carbazole compounds, on severe thrombocytopenia induced by nimustine (ACNU, 45 mg/kg administered for 2 consecutive days intravenously) in mice. Administration schedule studies revealed that pretreatment of mice with KF41399 was necessary to improve thrombocytopenia. Oral administration of KF41399 ameliorated thrombocytopenia induced by ACNU and accelerated the rate of platelet recovery in a dose-dependent fashion. In addition, KF41399 pretreatment improved the decrease in body weight and spleen weight and in the colony-forming activity of bone marrow mononuclear cells (MNC). Oral administration of KF41399 to normal mice induced G(0)/G(1)-phase accumulation of MNC as well as hematopoietic progenitor cells (lineage negative cells [Lin(-)]) and reduced the colony-forming activity of MNC. In Lin(-) cells derived from KF41399-treated mice, up-regulation of Bcl-2 and down-regulation of cyclin E and cyclin A proteins were observed. In the same cells, a decrease in the phosphorylated form of Rb protein and an increase in the p130 protein were observed without changes in the protein level of cell cycle dependent kinase 2 (Cdk2), Cdk4, and Cdk6. More important, KF41399 did not affect the antitumor activity of ACNU against mouse Sarcoma180 and human lung cancer LC 6. However, 25-mg/kg KF41399 treatment reduced the antitumor activity of ACNU against human lung cancer Lu-65, and 5 mg/kg KF41399 caused a slight reduction of the antitumor activity of ACNU without inducing thrombocytopenia. These results suggest that KF41399 might be useful as a chemoprotective agent to improve chemotherapy-induced thrombocytopenia and types of other toxicity. (Blood. 2000;95:3771-3780) PMID- 10845910 TI - Activated protein C reduces ischemia/reperfusion-induced renal injury in rats by inhibiting leukocyte activation. AB - We examined whether activated protein C (APC) reduces ischemia/reperfusion (I/R) induced renal injury by inhibiting leukocyte activation. In a rat model, intravenous administration of APC markedly reduced I/R-induced renal dysfunction and histological changes, whereas intravenous administration of dansyl glutamylglycylarginyl chloromethyl ketone-treated factor Xa (DEGR-FXa; active site-blocked factor Xa), heparin or diisopropyl fluorophosphate-treated APC (DIP APC; inactive derivative of ARC) had no effect. Furthermore, APC significantly inhibited the I/R-induced decrease in renal tissue blood flow and the increase in the vascular permeability, whereas neither DEGR-FXa, heparin, nor DIP-APC produced such effects. Renal I/R-induced increases in plasma levels of fibrin degradation products were significantly inhibited by APC, DEGR-FXa, and heparin. These observations suggest that APC reduces I/R-induced renal injury independently of its anticoagulant effects but in a manner dependent on its serine protease activity. Renal levels of tumor necrosis factor-alpha (TNF alpha), rat interleukin-8, and myeloperoxidase were significantly increased after renal I/R. These increases were significantly inhibited by APC but not by DEGR FXa, heparin, or DIP-APC. Leukocytopenia produced effects similar to those of APC. These findings strongly suggest that APC protects against I/R-induced renal injury not by inhibiting coagulation abnormalities but by inhibiting activation of leukocytes that play an important role in I/R-induced renal injury. Inhibition of leukocyte activation by APC could be explained by the inhibitory activity of TNF-alpha. (Blood. 2000;95:3781-3787) PMID- 10845911 TI - Cleaved high molecular weight kininogen binds directly to the integrin CD11b/CD18 (Mac-1) and blocks adhesion to fibrinogen and ICAM-1. AB - High molecular weight kininogen (HK) and its cleaved form (HKa) have been shown to bind to neutrophils. Based on studies using monoclonal antibodies (mAbs), we postulated that CD11b/CD18 (Mac-1) might be the receptor on the neutrophils for binding to HK/HKa. However, the direct interaction of HK/HKa and Mac-1 had not been demonstrated. We therefore transfected HEK 293 cells with human Mac-1. Cell binding assays using fluorescein isothiocyanate-labeled HKa showed increased binding to the Mac-1 transfected cells compared with the control transfected cells. The binding was specific because unlabeled HKa, Mac-1-specific antibody, and fibrinogen can inhibit the binding of biotin-HKa to Mac-1 transfected cells. HKa bound to Mac-1 transfected cells (20 000 molecules/cell) with a K(d) = 62 nmol/L. To demonstrate directly the formation of a complex between HKa and Mac-1, we examined the interaction of HKa and purified Mac-1 in a cell-free system using an IAsys resonant mirror optical biosensor. The association and dissociation rate constants (k(on) and k(off), respectively) were determined, and they yielded a dissociation constant (K(d)) of 3.2x10(-9) mol/L. The functional significance of direct interaction of HKa to Mac-1 was investigated by examining the effect of HKa on cellular adhesion to fibrinogen and intercellular adhesion molecule-1 (ICAM-1), molecules abundant in the injured vessel wall. HKa blocked the adhesion of Mac-1 transfected cells to fibrinogen and ICAM-1 in a dose-dependent manner. Thus, HKa may interrupt Mac-1-mediated cell-extracellular matrix and cell-cell adhesive interactions and may therefore influence the recruitment of circulating neutrophils/monocytes to sites of vessel injury. (Blood. 2000;95:3788-3795) PMID- 10845912 TI - Effect of recombinant von Willebrand factor reproducing type 2B or type 2M mutations on shear-induced platelet aggregation. AB - The aim was to better understand the function of von Willebrand factor (vWF) A1 domain in shear-induced platelet aggregation (SIPA), at low (200) and high shear rate (4000 seconds(-1)) generated by a Couette viscometer. We report on 9 fully multimerized recombinant vWFs (rvWFs) expressing type 2M or type 2B von Willebrand disease (vWD) mutations, characterized respectively by a decreased or increased binding of vWF to GPIb in the presence of ristocetin. We expressed 4 type 2M (-G561A, -E596K, -R611H, and -I662F) and 5 type 2B (rvWF-M540MM, -V551F, V553M, -R578Q, and -L697V). SIPA was strongly impaired in all type 2M rvWFs at 200 and 4000 seconds(-1). Decreased aggregation was correlated with ristocetin binding to platelets. In contrast, a distinct effect of botrocetin was observed, since type 2M rvWFs (-G561A, -E596K, and -I662F) were able to bind to platelets to the same extent as wild type rvWF (rvWF-WT). Interestingly, SIPA at 200 and 4000 seconds(-1) confirmed the gain-of-function phenotype of the 5 type 2B rvWFs. Our data indicated a consistent increase of SIPA at both low and high shear rates, reaching 95% of total platelets, whereas SIPA did not exceed 40% in the presence of rvWF-WT. Aggregation was completely inhibited by monoclonal antibody 6D1 directed to GPIb, underlining the importance of vWF-GPIb interaction in type 2B rvWF. Impaired SIPA of type 2M rvWF could account for the hemorrhagic syndrome observed in type 2M vWD. Increased SIPA of type 2B rvWF could be responsible for unstable aggregates and explain the fluctuant thrombocytopenia of type 2B vWD. (Blood. 2000;95:3796-3803) PMID- 10845913 TI - Synergistic induction of HLA class I expression by RelA and CIITA. AB - The major histocompatibility complex (MHC) class I genes are induced synergistically by interferons (IFN) and tumor necrosis factor (TNF), a response thought to involve the cooperative action of Rel/NF-kB and interferon regulatory factor (IRF) transcription factors. The IFN-gamma-inducible class II transcriptional activator (CIITA) has recently been shown to transactivate MHC class I as well as class II genes, and this investigation shows that CIITA synergizes strongly with RelA to stimulate HLA class I expression. The functional interaction of CIITA and RelA requires both promoter elements and the upstream Rel binding site and is not seen with a class II reporter. The promoter elements necessary for CIITA action are also required for induction by IFN-alpha. HLA-A and HLA-B loci respond differentially to IFNs, and we identify locus-specific differences in critical promoter elements in addition to known polymorphisms in the Rel and IRF binding sites. The HLA-A promoter is transactivated relatively poorly by CIITA and does not interact detectably with CREB proteins implicated in CIITA recruitment, but the synergism with RelA can compensate for this weakness. The present findings illustrate that multiple transcription factors cooperate to regulate class I expression and that their relative importance differs according to the locus and cell type examined. (Blood. 2000;95:3804-3808) PMID- 10845914 TI - Secretion of bioactive interleukin-1beta by dendritic cells is modulated by interaction with antigen specific T cells. AB - The role of interleukin-1beta (IL-1beta) as a regulator of the immune response, although extensively investigated, is still debated. We then studied the expression of IL-1beta by human dendritic cells (DCs), the professional antigen presenting cells, and its modulation during immune reactions in vitro. Our results show that, on maturation or tetanus toxoid presentation to specific CD4(+) CD40L(+) T lymphocytes, DCs begin to accumulate IL-1beta precursor (pro-IL 1beta) but do not secrete bioactive IL-1beta. In contrast, interaction with alloreactive T cells results in both stimulation of pro-IL-1beta synthesis and secretion of processed isoforms of the cytokine, that display biologic activity. Both CD4(+) and CD8(+) subsets of allospecific T lymphocytes are required: CD4(+) T cells drive the synthesis of pro-IL-1beta through CD40 engagement but have no effects on pro-IL-1beta processing; CD8(+) T cells, unable to induce synthesis of pro-IL-1beta per se, are responsible for the generation of mature IL-1beta by pro IL-1beta-producing DCs. Interleukin-1beta-converting enzyme (ICE) inhibitors do not prevent the recovery of IL-1beta bioactivity after allorecognition, indicating that allospecific CD8(+) T cells may induce the release of bioactive IL-1beta via mechanism(s) other than ICE activation. Altogether, these findings suggest that CD4(+) and CD8(+) T-lymphocyte subsets have distinct roles in the induction of IL-1beta secretion by DCs and support the hypothesis that IL-1beta plays a role in cell-mediated immune responses. (Blood. 2000;95:3809-3815) PMID- 10845915 TI - Selective disruption of interleukin 4 autocrine-regulated loop by a tyrosine kinase inhibitor restricts activity of T-helper 2 cells. AB - Interleukin (IL) 4 is a potent immunomodulatory cytokine secreted by T-helper 2 (Th2) cells and Th2 mast cells that promotes the commitment of cells. However, unregulated production and release of IL-4 can exacerbate allergic reactions and increase susceptibility to infectious organisms and viruses. Here, we present evidence that AG-490, a Janus tyrosine kinase (JAK) 2-JAK3 inhibitor, effectively blocked IL-4 gene expression and secretion in the Th2 cell line D10 that was not occurring after anti-CD3 antibody stimulation, whereas AG-490 had no inhibitory effect on production of other Th2 cytokines or cytokines synthesized by the corresponding Th1 cell line clone 29. AG-490 potently inhibited IL-4-mediated proliferation of both D10 and the IL-4-dependent cell line CT.4S. Moreover, AG 490 markedly inhibited IL-4 activation of JAK3 and blocked the downstream activation of signal transducer and activator of transcription 6, as judged by tyrosine phosphorylation, DNA binding, and transcription assays. In contrast, AG 490 did not affect tumor necrosis factor alpha activation of NF-kappaB at similar concentrations of drug. These data suggest that tyrosine kinase inhibitors that inhibit JAK3 may have previously unrecognized and selective clinical potential as immunotherapeutic drugs to treat Th2-mediated diseases driven by IL-4. (Blood. 2000;95:3816-3822) PMID- 10845916 TI - LPS induces apoptosis in macrophages mostly through the autocrine production of TNF-alpha. AB - The deleterious effects of lipopolysaccharide (LPS) during endotoxic shock are associated with the secretion of tumor necrosis factor (TNF) and the production of nitric oxide (NO), both predominantly released by tissue macrophages. We analyzed the mechanism by which LPS induces apoptosis in bone marrow-derived macrophages (BMDM). LPS-induced apoptosis reached a plateau at about 6 hours of stimulation, whereas the production of NO by the inducible NO-synthase (iNOS) required between 12 and 24 hours. Furthermore, LPS-induced early apoptosis was only moderately reduced in the presence of an inhibitor of iNOS or when using macrophages from iNOS -/-mice. In contrast, early apoptosis was paralleled by the rapid secretion of TNF and was almost absent in macrophages from mice deficient for one (p55) or both (p55 and p75) TNF-receptors. During the late phase of apoptosis (12-24 hours) NO significantly contributed to the death of macrophages even in the absence of TNF-receptor signaling. NO-mediated cell death, but not apoptosis induced by TNF, correlated with the induction of p53 and Bax genes. Thus, LPS-induced apoptosis results from 2 independent mechanisms: first and predominantly, through the autocrine secretion of TNF-alpha (early apoptotic events), and second, through the production of NO (late phase of apoptosis). (Blood. 2000;95:3823-3831) PMID- 10845917 TI - Increased apoptosis of peripheral blood T cells following allogeneic hematopoietic cell transplantation. AB - Lymphopenia and immune deficiency are significant problems following allogeneic hematopoietic cell transplantation (HCT). It is largely assumed that delayed immune reconstruction is due to a profound decrease in thymus-dependent lymphopoiesis, especially in older patients, but apoptosis is also known to play a significant role in lymphocyte homeostasis. Peripheral T cells from patients who received HCT were studied for evidence of increased cell death. Spontaneous apoptosis was measured in CD3(+) T cells following a 24-hour incubation using 7 amino-actinomycin D in conjunction with the dual staining of cell surface antigens. Apoptosis was significantly greater among CD3(+) T cells taken from patients 19-23 days after transplantation (30.4% +/- 12.5%, P <.05), and 1 year after transplantation (9.7% +/- 2.8%, P <.05) compared with healthy controls (4.0% +/- 1.5%). Increased apoptosis occurred preferentially in HLA (human leukocyte antigen)-DR positive cells and in both CD3(+)/CD4(+) and CD3(+)/CD8(+) T-cell subsets, while CD56(+)/CD3(-) natural killer cells were relatively resistant to apoptosis. The extent of CD4(+) T-cell apoptosis was greater in patients with grade II-IV acute graft-versus-host disease (GVHD) (33. 9% +/- 11.3%) compared with grade 0-I GVHD (14.6 +/- 6.5%, P <.05). T-cell apoptosis was also greater in patients who received transplantations from HLA-mismatched donors (39.5% +/- 10.4%, P <. 05) or HLA-matched unrelated donors (32.1% +/- 11.4%, P <.05) compared with patients who received transplantations from HLA-identical siblings (19.6% +/- 6.7%). The intensity of apoptosis among CD4(+) T cells was significantly correlated with a lower CD4(+) T-cell count. Together, these observations suggest that activation of T cells in vivo, presumably by alloantigens, predisposes the cells to spontaneous apoptosis, and this phenomenon is associated with lymphopenia. Activation-induced T-cell apoptosis may contribute to delayed immune reconstitution following HCT. (Blood. 2000;95:3832 3839) PMID- 10845918 TI - Synergistic effects of in vivo depletion of Ly-49A and Ly-49G2 natural killer cell subsets in the rejection of H2(b) bone marrow cell allografts. AB - Subsets of murine natural killer (NK) cells exist that express the Ly-49 family of molecules that recognize different major histocompatibility complex (MHC) determinants. Bone marrow transplantation studies were performed to examine the in vivo functions of 2 of these subsets. Subsets of Ly-49A and Ly-49G2 NK share specificity for the same MHC class 1 ligand, D(d), binding of which results in an inhibitory signal to the NK cell but allows them to lyse H2(b) targets in vitro. We therefore examined the ability of these subsets to reject H2(b) bone marrow cell allografts in lethally irradiated mice. Surprisingly, depletion of Ly-49A(+) NK cells in BALB/c or B10.D2 mice (both H2(d)) had no effect on the rejection of H2(b) BMC. However, Ly-49A depletion did partially abrogate the ability of B10.BR (H2(k)) mice to reject H2(b) allografts. Although depletion of either Ly-49A(+) or Ly-49G2(+) NK cells alone had no effect on the ability of B10.D2 mice to reject H2(b) BMC, depletion of both subsets dramatically and synergistically abrogated rejection. Studies with various B10 congenic mice and their F(1) hybrids indicate that this synergy between Ly49A and Ly4G2 depletion occurs in every instance. Thus, Ly-49A(+) NK cells appear to play a role in the rejection H2(b) bone marrow allografts, but, in most strains of mice studied, Ly-49G2(+) NK cells must also be eliminated. The putative roles of these NK cell subsets in clinical transplantation remains to be elucidated. (Blood. 2000;95:3840-3844) PMID- 10845919 TI - Lineage-specific regulation of the murine RAG-2 promoter: GATA-3 in T cells and Pax-5 in B cells. AB - Recombination activating gene-1 (RAG-1) and RAG-2 are expressed in lymphoid cells undergoing the antigen receptor gene rearrangement. A study of the regulation of the mouse RAG-2 promoter showed that the lymphocyte-specific promoter activity is conferred 80 nucleotide (nt) upstream of RAG-2. Using an electrophoretic mobility shift assay, it was shown that a B-cell-specific transcription protein, Pax-5, and a T-cell-specific transcription protein, GATA-3, bind to the -80 to -17 nt region in B cells and T cells, respectively. Mutation of the RAG-2 promoter for Pax-5- and GATA-3-binding sites results in the reduction of promoter activity in B cells and T cells. These results indicate that distinct DNA binding proteins, Pax-5 and GATA-3, may regulate the murine RAG-2 promoter in B and T lineage cells, respectively. (Blood. 2000;95:3845-3852) PMID- 10845920 TI - CD40 activation does not protect chronic lymphocytic leukemia B cells from apoptosis induced by cytotoxic T lymphocytes. AB - Cytotoxic T lymphocytes (CTLs) can kill target cells by the granule/exocytosis pathway or the Fas-mediated apoptosis pathway. The sensitivity of chronic lymphocytic leukemia (CLL) B cells to CTL-mediated apoptosis before and after CD40 activation was examined. Resting or CD40-activated CLL cells were found to be equally sensitive to class I-restricted CTL-mediated killing. Despite expressing CD95, the CD40-activated CLL target cells were found to be resistant to apoptosis induced by CH11, an IgM CD95 monoclonal antibody (mAb). Consistent with this, inhibitors of caspases, which are involved in the Fas-induced apoptotic pathway (eg, N-carbobenzoxy-Val-Ala-Asp fluoromethyl ketone [z-VAD fmk]), were unable to block destruction of CLL target cells by CTL. In addition, preincubation of the effector T cells with the anti-Fas ligand mAb NOK-2 failed to inhibit their subsequent ability to kill CLL target cells. On the other hand, CTL activity was blocked by inhibitors of the granule exocytosis pathway such as ethylene-glyco-tetra-acetic acid or concanamycin A. These results indicate that CD40 activation does not impair the sensitivity of CLL cells to Fas-independent CTL-mediated apoptosis. (Blood. 2000;95:3853-3858) PMID- 10845921 TI - A(2A) receptor dependent and A(2A) receptor independent effects of extracellular adenosine on murine thymocytes in conditions of adenosine deaminase deficiency. AB - Adenosine deaminase (ADA) deficiency causes severe combined immunodeficiency (SCID) and is accompanied by T-cell depletion and accumulation of both intracellular and extracellular adenosine (extAdo) and deoxyadenosine. To better understand the causes of T-cell depletion in vivo and to discriminate between extracellular and intracellular effects of exogenously added adenosine in vitro, we investigated mechanisms of 2 different effects of adenosine on murine thymocytes. These effects of adenosine include direct induction of apoptosis in about 6% to 15% thymocytes and inhibition of T-cell receptor (TCR)-induced activation of the majority of thymocytes with inhibited ADA. A(2A) adenosine receptors, but not A(2B), A(1), or A(3) receptors, are shown to be mostly responsible for extAdo-triggered signaling (cyclic adenosine monophosphate [cAMP] accumulation) in murine thymocytes and this prompted studies of the effects of extAdo on thymocytes from A(2A)R gene-deficient mice. It is found that direct apoptotic effects of extAdo on CD4(+)CD8(+) double positive (DP) thymocytes are completely accounted for by signaling through A(2A)R, with no contribution of intracellular lymphotoxicity or of compensating A(2B)Rs because only A(2A)R +/+, but not A(2A)R -/- thymocytes were susceptible to apoptotic effects of extAdo. Studies of the effects of cAMP-raising agents support observations of extAdo/A(2A)R/cAMP-triggered apoptosis in DP thymocytes. Unexpectedly, the extAdo strongly inhibited TCR-triggered activation of both A(2A)R +/+ and A(2A)R -/- thymocytes in the presence of ADA inhibitors. This was confirmed with thymocytes from ADA gene-deficient mice, suggesting the existence of A(2A)R-independent effects of extAdo on thymocytes. The presented data raises questions about the identity and functional role of A(2A)R-expressing thymocytes in T-cell differentiation and of the role of TCR-antagonizing effects of extAdo in conditions of ADA SCID. (Blood. 2000;95:3859-3867) PMID- 10845922 TI - Engagement of CD11b and CD11c beta2 integrin by antibodies or soluble CD23 induces IL-1beta production on primary human monocytes through mitogen-activated protein kinase-dependent pathways. AB - beta2 integrins are involved in the recruitment of leukocytes to inflammatory sites and in cellular activation. We demonstrate that ligation of CD11b (Mac-1, CR3) or CD11c (p150, CR4) alpha chains of beta2 integrins by mAbs or soluble chimeric CD23 (sCD23) on human freshly isolated monocytes rapidly stimulates high levels of interleukin-1beta production. This induction takes place at the transcriptional level and is regulated by members of the mitogen-activated protein kinase (MAPK) family. Indeed, stimulation of monocytes through engagement of CD11b or CD11c results in the phosphorylation and activation of ERK1, ERK2, and p38/SAPK2 MAP kinases. U0126, a potent inhibitor of the upstream activator of ERK1/2, ie, MEK1/2, suppresses IL-1beta messenger RNA (mRNA) expression in a dose dependent fashion, showing the implication of this pathway in the transcriptional control of IL-1beta production. On the other hand, inhibition of p38 by SB203580 indicates that this MAPK is involved in the control of IL-1beta production at both transcriptional and translational levels. Together these data demonstrate that ligation of CD11b and CD11c beta2 integrins by mAbs or sCD23 fusion proteins triggers the activation of 2 distinct MAPK signaling pathways that cooperate in controlling IL-1beta synthesis at different levels. (Blood. 2000;95:3868-3877) PMID- 10845923 TI - Salivary gland mucosa-associated lymphoid tissue lymphoma immunoglobulin V(H) genes show frequent use of V1-69 with distinctive CDR3 features. AB - Salivary gland mucosa associated lymphoid tissue (MALT) type lymphomas are B-cell neoplasms that develop out of a reactive infiltrate, often associated with Sjogren's syndrome. Previous reports from our laboratory involving 10 patients suggested these lymphomas expressed a restricted immunoglobulin (Ig) V(H) gene repertoire with over use of V1-69 gene segments. To better determine the frequency of V1-69 use and whether there may also be selection for CDR3 structures, we sequenced the V(H) genes from 15 additional cases. Over half of the potentially functional V(H) genes (8 of 14) used a V(H)1 family V1-69 gene segment, whereas the other cases used different gene segments from the V(H)1 (V1 46), V(H)3 (V3-7, V3-11, V3-30.3, V3-30.5), and V(H)4 (V4-39) families. The 8 V1 69 V(H) genes used 5 different D segments in various reading frames, but all used a J4 joining segment. The V1-69 CDR3s showed remarkable similarities in lengths (12-14 amino acids) and stretches of 2 to 3 amino acids between the V-D and D-J junctions. They did not resemble CDR3s typical of V1-69 chronic lymphocytic leukemias. This study extends our earlier work in establishing that salivary gland MALT lymphomas represent a highly selected B-cell population. Frequent use of V1-69 appears to differ from MALT lymphomas that develop at other sites. The high degree of CDR3 similarity among the V1-69 cases suggests that different salivary gland lymphomas may bind similar, if not identical epitopes. Although the antigen specificities are presently unknown, similar characteristic CDR3 sequences are often seen with V1-69 encoded antibodies that have anti-IgG or rheumatoid factor activity. (Blood. 2000;95:3878-3884) PMID- 10845924 TI - BCL10 gene mutation in lymphoma. AB - BCL10 is directly involved in t(1;14)(p22;q32) of mucosa-associated lymphoid tissue (MALT) lymphoma. Wild-type BCL10 promoted apoptosis and suppressed malignant transformation in vitro, whereas truncated mutants lost the pro apoptotic activity and exhibited gain of function enhancement of transformation. We studied 220 lymphomas for genomic BCL10 mutation by polymerase chain reaction single-strand conformational polymorphism and DNA sequencing. Nineteen mutations were found in 13 lymphoma specimens, as follows: 8 of 120 (6.7%) mucosa associated lymphoid tissue (MALT) lymphomas, 4 of 42 (9.5%) follicular lymphomas, and 1 of 23 (4.3%) diffuse large B-cell lymphomas. No mutations were found in 14 mantle cell lymphomas or 21 T-cell lymphomas. High-grade MALT lymphoma tended to show a slightly higher mutation frequency (2 of 25, 8%) than low-grade MALT tumor (6 of 95, 6.3%). Among low-grade gastric MALT lymphoma, mutations were found in 3 of 11 tumors that did not respond to Helicobacter pylori eradication therapy, but none were found in 22 tumors that regressed completely after H pylori eradication. All 14 potentially pathogenic mutations were distributed in the carboxyl terminal domain of BCL10. Deletion accounted for 10 of these mutations; 10 of 14 mutations caused truncated forms of BCL10. Western blot analysis of a mutant case confirmed the presence of truncated BCL10 products of anticipated size. Our results suggest that BCL10 mutation may play a pathogenic role in B cell lymphoma development, particularly in aggressive and antibiotic unresponsive MALT lymphomas, and may further implicate the biologic importance of the carboxyl terminal of the molecule. (Blood. 2000;95:3885-3890) PMID- 10845925 TI - TEL-JAK2 transgenic mice develop T-cell leukemia. AB - We previously reported a fusion between TEL and JAK2 in a t(9;12)(p24;p13) chromosomal translocation in childhood acute T-cell leukemia. This fusion gene encodes a TEL-JAK2 chimeric protein in which the 336 amino-terminal residues of TEL, including its specific self-association domain, are fused to the kinase domain of JAK2. TEL-JAK2 exhibits constitutive activation of its tyrosine kinase activity which, in turn, confers growth factor-independent proliferation to the interleukin-3-dependent Ba/F3 hematopoietic cell line. To elucidate the properties of TEL-JAK2 in primary cells and to create an animal model for TEL JAK2-induced leukemia, we generated transgenic mice in which the TEL-JAK2 complementary DNA was placed under the transcriptional control of the EmuSRalpha enhancer/promoter. TEL-JAK2 founder mice and their transgenic progeny developed fatal leukemia at 4 to 22 weeks of age. Selective amplification of CD8-positive T cells was observed in blood, lymph nodes, thymus, spleen, and bone marrow. Expression of a tyrosine-phosphorylated TEL-JAK2 protein and activation of STAT1 and STAT5 (signal transducer and activator of transcription) were detected in leukemic tissues. TEL-JAK2 diseased mice also displayed invasion of nonhematopoietic organs, including liver, brain, lung, and kidney, by leukemic T cells. Leukemic organs of founder and transgenic progeny contained a monoclonal/oligoclonal T-cell population as analyzed by the rearrangement of the TCRbeta locus. Transplantation of TEL-JAK2 leukemic cells in nude mice confirmed their invasive nature. We conclude that the TEL-JAK2 fusion is an oncogene in vivo and that its expression in lymphoid cells results in the preferential expansion of CD8-positive T cells. (Blood. 2000;95:3891-3899) PMID- 10845926 TI - Biologic response of B lymphoma cells to anti-CD20 monoclonal antibody rituximab in vitro: CD55 and CD59 regulate complement-mediated cell lysis. AB - The chimeric anti-CD20 MAb rituximab has recently become a treatment of choice for low-grade or follicular non-Hodgkin's lymphomas (FL) with a response rate of about 50%. In this report, we have investigated the mechanism of action of rituximab on 4 FL and 1 Burkitt's lymphoma (BL) cell lines, 3 fresh FL samples and normal B cells in vitro. Rituximab efficiently blocks the proliferation of normal B cells, but not that of the lymphoma lines. We did not detect significant apoptosis of the cell lines in response to rituximab alone. All cell lines were targets of antibody-dependent cellular cytotoxicity (ADCC). On the other hand, human complement-mediated lysis was highly variable between cell lines, ranging from 100% lysis to complete resistance. Investigation of the role of the complement inhibitors CD35, CD46, CD55, and CD59 showed that CD55, and to a lesser extent CD59, are important regulators of complement-mediated cytotoxicity (CDC) in FL cell lines as well as in fresh cases of FL: Blocking CD55 and/or CD59 function with specific antibodies significantly increased CDC in FL cells. We conclude that CDC and ADCC are major mechanisms of action of rituximab on B-cell lymphomas and that a heterogeneous susceptibility of different lymphoma cells to complement may be at least in part responsible for the heterogeneity of the response of different patients to rituximab in vivo. Furthermore, we suggest that the relative levels of CD55 and CD59 may become useful markers to predict the clinical response. (Blood. 2000;95:3900-3908) PMID- 10845927 TI - Ki-4(scFv)-ETA', a new recombinant anti-CD30 immunotoxin with highly specific cytotoxic activity against disseminated Hodgkin tumors in SCID mice. AB - The human lymphocyte activation marker CD30 is highly overexpressed on Hodgkin/Reed-Sternberg cells and represents an ideal target for selective immunotherapy. We used the murine anti-CD30 hybridoma Ki-4 to construct a new recombinant immunotoxin (rIT) for possible clinical use in patients with CD30(+) lymphoma. Hybridoma V genes were polymerase chain reaction-amplified, assembled, cloned, and expressed as a mini-library for display on filamentous phage. Functional Ki-4 scFv obtained by selection of binding phage on the CD30 expressing Hodgkin lymphoma cell line L540cy was inserted into the bacterial expression vector pBM1.1 and fused to a deletion mutant of Pseudomonas exotoxin A (ETA'). Periplasmically expressed Ki-4(scFv)-ETA' demonstrated specific activity against a variety of CD30(+) lymphoma cells as assessed by different in vitro assays. To evaluate in vivo antitumor activity, severe combined immunodeficient mice challenged with human lymphoma cell lines were treated with the immunotoxin. The blood distribution time t(1/2)alpha of Ki-4(scFv)-ETA' was 19 minutes, and its serum elimination time t(1/2)alpha was 193 minutes. A single intravenous injection of 40 microg rIT 1 day after tumor inoculation rendered 90% of the mice tumor free, extending the mean survival time to more than 200 days compared with 38.1 days in the phosphate-buffered saline control group (P <.001). This new rIT is a promising candidate for further clinical evaluation in patients with Hodgkin lymphoma or other CD30(+) malignancies. (Blood. 2000;95:3909-3914) PMID- 10845928 TI - Constitutive activation of transcription factor AP-1 in primary adult T-cell leukemia cells. AB - Human T-cell leukemia virus type-I (HTLV-I) is the etiologic agent of adult T cell leukemia (ATL). This study examined the status of the oncogenic transcription factor AP-1 in leukemic cells freshly isolated from patients with ATL. Leukemic cells from peripheral blood of all patients with ATL exhibited constitutive AP-1 DNA binding activity, whereas mononuclear cells from normal individuals did not. In agreement with previous studies, HTLV-I transforming protein, Tax, was found to stimulate the DNA binding activity of AP-1 in a T-cell line. However, HTLV-I genes, including Tax, were not significantly expressed in leukemic cells freshly obtained from patients with ATL. Moreover, all T-cell lines derived from leukemic cells of patients with ATL also displayed constitutive AP-1 DNA binding activity, but expressed little Tax protein. Thus, leukemic cells of patients with ATL appear to have Tax-independent mechanisms that induce AP-1 activity, both in vivo and in vitro. In antibody supershift experiments, AP-1 in fresh leukemic cells and ATL-derived cell lines were found to contain JunD. Consistently, all primary ATL cells and ATL-derived cell lines expressed high levels of JunD messenger RNA. Our results suggest that AP-1 is activated in leukemic cells of patients with ATL through a Tax-independent mechanism and this may play a role in the deregulated phenotypes of ATL leukemic cells. (Blood. 2000;95:3915-3921) PMID- 10845929 TI - Primary cutaneous follicle center cell lymphoma with follicular growth pattern. AB - Cutaneous B-cell infiltrates showing a prominent follicular growth pattern with germinal centers are thought by some authors to represent either marginal zone lymphomas with reactive germinal centers or pseudolymphomas. To establish whether a true primary cutaneous follicular lymphoma exists, we studied biopsies from 15 patients with skin lesions characterized histopathologically by the presence of B cell infiltrates with follicular pattern. Staging investigations, including bone marrow biopsy, were negative in all patients. All were negative for bcl-2 protein expression and did not present the t(14;18). In all biopsy specimens neoplastic follicles showed 1 or more morphologic or immunophenotypic criteria of malignancy (presence of a reduced mantle zone, absence of tingible body macrophages, reduced proliferation rate). In 9 specimens a monoclonal rearrangement of J(H) genes could be detected by polymerase chain reaction analysis. After laser beam microdissection, a band of the same length could be observed in 6 probes from different follicles from the same specimen, indicating the presence of the same monoclonal population of follicle center cells. Follow-up examinations in all patients revealed no evidence of extracutaneous spread (mean follow-up, 48.7 months). Our study demonstrates that primary cutaneous follicular lymphoma represents a distinct entity of the cutaneous B-cell lymphomas. (Blood. 2000;95:3922-3928) PMID- 10845930 TI - Liposomal Bcl-2 antisense oligonucleotides enhance proliferation, sensitize acute myeloid leukemia to cytosine-arabinoside, and induce apoptosis independent of other antiapoptotic proteins. AB - The antiapoptotic proteins, Bcl-2 and Bcl-X(L), are expressed in most cases of acute myeloid leukemia (AML) and may contribute to drug resistance in AML. We tested the hypothesis that down-regulation of Bcl-2 alone by antisense oligodeoxynucleotides (Bcl-2-AS) induces apoptosis, even in the presence of other antiapoptotic genes. We tested Bcl-2-AS in myeloid leukemic HL-60 cells, in Bcl-2 and Bcl-X(L) overexpressing HL-60-DOX cells, and in primary AML samples. Down regulation of Bcl-2 by Bcl-2-AS reduced the viability of HL-60 cells and, less effectively, HL-60-DOX cells and increased ara-C cytotoxicity in both cell lines. Incubation of primary AML blasts with Bcl-2-AS decreased Bcl-2 expression in CD34(+) blast cells after induction of apoptosis and enhancement of ara-C cytotoxicity in 11 of 19 primary AML samples. In 8 samples in which Bcl-2-AS did not induce apoptosis, baseline Bcl-2 levels were found to be strikingly high. The expression of other antiapoptotic proteins (Bcl-X(L), Bag-1, A1, and Mcl-1) did not prevent Bcl-2-AS-induced apoptosis. Bcl-2-AS also inhibited colony formation of AML progenitor cells. Low concentrations of Bcl-2-AS induced significant increases in S-phase cells (P =.04). Results establish Bcl-2 as a critical target for AS strategies in AML in which the baseline levels predict response to Bcl-2 AS. Bcl-2 exerts both antiapoptotic and antiproliferative functions in AML. Because early normal hematopoietic stem cells do not express Bcl-2, Bcl-2-AS therapy should be highly selective for AML cells. (Blood. 2000;95:3929-3938) PMID- 10845932 TI - Differential expression of a novel C-terminally truncated splice form of SMAD5 in hematopoietic stem cells and leukemia. AB - SMADs are evolutionarily conserved transducers of the differentiation and growth arrest signals from the transforming growth factor/BMP (TGF/BMP) family of ligands. Upon receptor activation, the ligand-restricted SMADs(1-35) are phosphorylated in the C-terminal MH2 domain and recruit the common subunit SMAD4/DPC-4 gene to the nucleus to mediate target gene expression. Frequent inactivating mutations of SMAD4, or less common somatic mutations of SMAD2 seen in solid tumors, suggest that these genes have a suppressor function. However, there have been no identified mutations of SMAD5, although the gene localizes to the critical region of loss in chromosome 5q31.1 (chromosome 5, long arm, region 3, band 1, subband 1) in myelodysplasia (MDS) and acute myelogenous leukemia (AML). A ubiquitously expressed novel isoform, SMAD5beta, encodes a 351 amino acid protein with a truncated MH2 domain and a unique C-terminal tail of 18 amino acids, which may be the functional equivalent of inactivating mutations. The levels of SMAD5beta transcripts are higher in the undifferentiated CD34(+) hematopoietic stem cells than in the terminally differentiated peripheral blood leukocytes, thereby implicating the beta form in stem cell homeostasis. Yeast 2 hybrid interaction assays reveal the lack of physical interactions between SMAD5beta and SMAD5 or SMAD4. The expression of SMAD5beta may represent a novel mechanism to protect pluripotent stem cells and malignant cells from the growth inhibitory and differentiation signals of BMPs. (Blood. 2000;95:3945-3950) PMID- 10845931 TI - p53 stabilization and functional impairment in the absence of genetic mutation or the alteration of the p14(ARF)-MDM2 loop in ex vivo and cultured adult T-cell leukemia/lymphoma cells. AB - Human T-cell lymphotropic virus type I (HTLV-I) transforms T cells in vitro, and the viral transactivator Tax functionally impairs the tumor suppressor p53 protein, which is also stabilized in HTLV-I-infected T cells. Thus, the functional impairment of p53 is essential to maintain the viral-induced proliferation of CD4+ mature T cells. However, in the CD4+ leukemic cells of patients with adult T-cell leukemia/lymphoma (ATLL), the viral transactivator does not appear to be expressed, and p53 mutations have been found only in a fraction of patients. We sought to investigate whether p53 function is impaired, in ex vivo samples from patients with ATLL, in the absence of genetic mutations. Here we demonstrate that the p53 protein is stabilized also in ex vivo ATLL samples (10 of 10 studied) and that at least in 2 patients p53 stabilization was not associated with genetic mutation. Furthermore, the assessment of p53 function after ionizing radiation of ATLL cells indicated an abnormal induction of the p53 responsive genes GADD45 and p21(WAF1) in 7 of 7 patients. In 2 of 2 patients, p53 regulation of cell-cycle progression appeared to be impaired as well. Because p53 is part of a regulatory loop that also involves MDM2 and p14(ARF), the status of the latter proteins was also assessed in cultured or fresh ATLL cells. The p97 MDM2 protein was not detected by Western blot analysis in established HTLV-I infected T-cell lines or ex vivo ATLL cell lysates. However, the MDM2 protein could be easily detected after treatment of cells with the specific proteasome inhibitor lactacystin, suggesting a normal regulation of the p53-MDM2 regulating loop. Similarly, p14(ARF) did not appear to be aberrantly expressed in ex vivo ATLL cells nor in any of the established HTLV-I-infected T-cell lines studied. Thus, p53 stabilization in HTLV-I infection occurs in the absence of genetic mutation and alteration of the physiologic degradation pathway of p53. (Blood. 2000;95:3939-3944) PMID- 10845933 TI - Estradiol-stimulated nitric oxide release in human granulocytes is dependent on intracellular calcium transients: evidence of a cell surface estrogen receptor. AB - We tested the hypothesis that estrogen acutely stimulates constitutive nitric oxide synthase activity in human granulocytes by acting on a cell surface estrogen receptor (ER). The release of nitric oxide was measured in real time with an amperometric probe. Exposure of granulocytes to 17beta-estradiol stimulated NO release within seconds in a concentration-dependent manner. The NO release was also stimulated by 17beta-estradiol conjugated to bovine serum albumin (E(2)-BSA), which suggests mediation by a cell surface receptor. Tamoxifen, an ER inhibitor, antagonized the action of both 17beta-estradiol and E(2)-BSA, whereas ICI 182,780, an inhibitor of the nuclear ER, had no effect. Using dual emission microfluorometry in a calcium-free medium, the 17beta estradiol-stimulated release of NO from granulocytes was shown to be dependent on intracellular calcium ([Ca(2+)]i) transients in a tamoxifen-sensitive process. Exposure to BAPTA-AM (1,2bis-(-aminophenoxy)ethans-N,N,N', N'-tetraacetic acid tetra(acetoxyymethyl) ester), a [Ca(2+)]i chelator, reduced [Ca(2+)]i in response to E(2)-BSA, and depleting [Ca(2+)]i stores abolished the effect of 17beta estradiol on NO release. Confocal photomicrographs using E(2)-BSA-FITC (fluorescein isothiocyanate) revealed cell membrane reactivity. Estrogen stimulated NO release had an immunosuppressive effect, and it initiated granulocyte rounding and loss of adherence in a tamoxifen-sensitive manner. Finally, using reverse transcriptase-polymerase chain reaction, human neutrophil granulocytes expressed ERalpha but not ERbeta, suggesting that ERalpha may be the membrane receptor for 17beta-estradiol. The study demonstrated that a physiological dose of estrogen down-regulates granulocyte activity by acutely stimulating NO release via the activation of a cell surface ER which is coupled to increases in [Ca(2+)]i. (Blood. 2000;95:3951-3958) PMID- 10845934 TI - Constitutive activation of the MAPK pathway mediates v-fes-induced mitogenesis in murine macrophages. AB - Fes is a nonreceptor tyrosine kinase expressed at the highest level in macrophages. We previously showed that the overexpression of c-fes in murine macrophages of the BAC-1.2F5 cell line renders these cells independent of macrophage colony-stimulating factor (MCSF) for survival and proliferation, although no direct relationship could be established between tyrosine phosphorylated substrates of Fes- and MCSF receptor-dependent signaling and mitogenesis. In this study, we investigated whether the mitogen-activated protein kinase (MAPK) pathway is involved in the growth factor-independent growth of v fes-overexpressing macrophages. We found a constitutively increased phosphorylation of extracellularly regulated kinase (ERK) in v-fes-overexpressing macrophages as compared with mock-infected cells. This finding was associated with activation of mitogen/extracellular signal-regulated kinase (MEK) and with constitutive localization of ERK in the nucleus. Treatment of v-fes overexpressing cells with the MEK-specific inhibitor PD98059 markedly reduced cell growth, hyperphosphorylation, and nuclear localization of ERK, indicating that the MAPK pathway mediates the mitogenic effect of v-fes. (Blood. 2000;95:3959-3963) PMID- 10845935 TI - Hepatocyte growth factor-stimulated invasiveness of monocytes. AB - Hepatocyte growth factor (HGF) is a pluripotent cytokine with mitogenic, motogenic, and morphogenic activity for mainly epithelial and endothelial target cells. We previously demonstrated that the specific HGF receptor, MET, is induced in stimulated peripheral blood monocytes. In this study, we analyzed the functional consequences of MET activation in primary cultures of peripheral blood monocytes from healthy donors. After stimulation of MET-expressing monocytes with recombinant HGF, the gene-expression profile of peripheral blood mononuclear cells and monocytes was significantly modulated, especially with regard to genes involved in cell movement. After stimulation of primary cultured monocytes with HGF, invasion assays showed a significantly increased matrigel invasion rate that was completely abolished by neutralizing antibodies to HGF. The HGF-activated invasiveness and the altered gene-expression profile suggest a proinflammatory role for HGF stimulation of monocytes and support the hypothesis that the HGF/MET signaling system plays an important part in the activation of the nonspecific cellular inflammatory response. (Blood. 2000;95:3964-3969) PMID- 10845936 TI - Posttranscriptional cell cycle-dependent regulation of human FANCC expression. AB - The Fanconi Anemia (FA) Group C complementation group gene (FANCC) encodes a protein, FANCC, with a predicted M(r) of 63,000 daltons. FANCC is found in both the cytoplasmic and the nuclear compartments and interacts with certain other FA complementation group proteins as well as with non-FA proteins. Despite intensive investigation, the biologic roles of FANCC and of the other cloned FA gene products (FANCA and FANCG) remain unknown. As an approach to understanding FANCC function, we have studied the molecular regulation of FANCC expression. We found that although FANCC mRNA levels are constant throughout the cell cycle, FANCC is expressed in a cell cycle-dependent manner, with the lowest levels seen in cells synchronized at the G1/S boundary and the highest levels in the M-phase. Cell cycle-dependent regulation occurred despite deletion of the 5' and 3' FANCC untranslated regions, indicating that information in the FANCC coding sequence is sufficient to mediate cell cycle-dependent regulation. Moreover, inhibitors of proteasome function blocked the observed regulation. We conclude that FANCC expression is controlled by posttranscriptional mechanisms that are proteasome dependent. Recent work has demonstrated that the functional activity of FA proteins requires the physical interaction of at least FANCA, FANCC, and FANCG, and possibly of other FA and non-FA proteins. Our observation of dynamic control of FANCC expression by the proteasome has important implications for understanding the molecular regulation of the multiprotein complex. (Blood. 2000;95:3970-3977) PMID- 10845937 TI - Mild spherocytic hereditary elliptocytosis and altered levels of alpha- and gamma adducins in beta-adducin-deficient mice. AB - The membrane skeleton, a dynamic network of proteins associated with the plasma membrane, determines the shape and mechanical properties of erythrocytes. Deficiencies or defects in membrane skeletal proteins are associated with inherited disorders of erythrocyte morphology and function. Adducin is one of the proteins localized at the spectrin-actin junction of the membrane skeleton. In this work we show that deficiency of beta-adducin produces an 80% decrease of alpha-adducin and a fourfold up-regulation of gamma-adducin in erythrocytes. beta Adducin or any other isoform generated by translation of abnormally spliced messenger RNAs could not be detected by our antibodies either in ghosts or in cytoplasm of -/- erythrocytes. Actin levels were diminished in mutant mice, suggesting alterations in the actin-spectrin junctional complexes due to the absence of adducin. Elliptocytes, ovalocytes, and occasionally spherocytes were found in the blood film of -/- mice. Hematological values showed an increase in reticulocyte counts and mean corpuscular hemoglobin concentration, decreased mean corpuscular volume and hematocrit, and normal erythrocyte counts that, associated to splenomegaly, indicate that the mice suffer from mild anemia with compensated hemolysis. These modifications are due to a loss of membrane surface and dehydration that result in an increase in the osmotic fragility of red blood cells. The marked alteration in osmotic fragility together with the predominant presence of elliptocytes is reminiscent of the human disorder called spherocytic hereditary elliptocytosis. Our results suggest that the amount of adducin remaining in the mutant animals (presumably alphagamma adducin) could be functional and might account for the mild phenotype. (Blood. 2000;95:3978-3985) PMID- 10845938 TI - Detection of active hepatitis C virus and hepatitis G virus/GB virus C replication in bone marrow in human subjects. AB - We have analyzed the presence of hepatitis C virus (HCV) and hepatitis G virus (HGV) sequences in bone marrow and serum samples from 48 patients of a hematologic outpatient clinic. HCV RNA was detected in 18 (38%) and 15 (31%) and HGV RNA was detected in 6 (13%) and 9 (19%) of serum and bone marrow samples, respectively. In 3 patients, HGV RNA was detectable in bone marrow but not in the serum; 2 of these patients were negative for the presence of specific antibodies. Using a highly strand-specific Tth-based reverse transcriptase-polymerase chain reaction (RT-PCR), the presence of HCV RNA and HGV RNA negative strand was demonstrated in 4 and 5 bone marrow samples, respectively. Our study shows that HCV and HGV can replicate in bone marrow; in the case of HGV, analysis of serum may underestimate the true prevalence of infection. (Blood. 2000;95:3986-3989) PMID- 10845939 TI - Longitudinal monitoring of immune reconstitution by CDR3 size spectratyping after T-cell-depleted allogeneic bone marrow transplant and the effect of donor lymphocyte infusions on T-cell repertoire. AB - Delayed immune reconstitution after allogeneic bone marrow transplantation (BMT) with associated infection is a major cause of morbidity and mortality. We used third complementarity region (CDR3) size spectratyping as a tool for monitoring T cell repertoire reconstitution in 19 patients over a median time of 40 months after T-cell-depleted allogeneic BMT for chronic myeloid leukemia (CML). Furthermore, the effect of donor lymphocyte infusions (DLI) for the treatment of relapse in 18 of the 19 patients was analyzed. All BMT recipients had irregular spectratypes in the first 3- to -6 months after transplant. These evolved to more normal patterns by 12 months after transplant and continued to improve thereafter. In approximately a third of the patients, it took 2 to 3 years for all spectratypes to normalize, whereas in the other two thirds, some abnormal spectratypes persisted even after several years. In 9 patients, there was no immediate change in the CDR3 size profiles after DLI. In 3 patients, spectratypes improved slightly after DLI, whereas in 6 patients, spectratypes became more restricted and irregular. Overall, T-cell spectratypes in BMT patients were characterized by instability over time and in patients with graft-versus-host disease (GVHD), this was even more exaggerated. Several factors, such as pre-BMT conditioning, T-cell depletion of the donor marrow, loss of thymic function in adults, exposure to infectious agents, GVHD, and immunosuppressive treatment, are likely contributors to the delay in T-cell-repertoire reconstitution. (Blood. 2000;95:3990-3995) PMID- 10845940 TI - T-cell depletion of bone marrow transplants for leukemia from donors other than HLA-identical siblings: advantage of T-cell antibodies with narrow specificities. AB - T-cell depletion of donor marrow decreases graft-versus-host disease resulting from transplants from unrelated and human leukocyte antigen (HLA)-mismatched related donors. However, there are diverse strategies for T-cell-depleted transplantation, and it is uncertain whether any improve leukemia-free survival (LFS). To compare strategies for T-cell-depleted alternative donor transplants and to compare T-cell depleted with non-T-cell-depleted transplants, we studied 870 patients with leukemia who received T-cell-depleted transplants from unrelated or HLA-mismatched related donors from 1982 to 1994. Outcomes were compared with those of 998 non-T-cell-depleted transplants. We compared LFS using different strategies for T-cell-depleted transplantation considering T-cell depletion technique, intensity of pretransplant conditioning, and posttransplant immune suppression using proportional hazards regression to adjust for other prognostic variables. Five categories of T-cell depletion techniques were considered: narrow-specificity antibodies, broad-specificity antibodies, Campath antibodies, elutriation, and lectins. Strategies resulting in similar LFS were pooled to compare T-cell-depleted with non-T-cell-depleted transplants. Recipients of transplants T-cell depleted by narrow-specificity antibodies had lower treatment failure risk (higher LFS) than recipients of transplants T-cell depleted by other techniques. Compared with non-T-cell-depleted transplants (5 year probability +/- 95% confidence interval [CI] of LFS, 31% +/- 4%), 5-year LFS was 29% +/- 5% (P = NS) after transplants T-cell depleted by narrow-specificity antibodies and 16% +/- 4% (P <.0001) after transplants T-cell depleted by other techniques. After alternative donor transplantation, T-cell depletion of donor marrow by narrow-specificity antibodies resulted in LFS rates that were higher than those for transplants T-cell depleted using other techniques but similar to those for non-T-cell-depleted transplants. (Blood. 2000;95:3996-4003) PMID- 10845941 TI - Megakaryocyte dense granule components are sorted in multivesicular bodies. AB - Recent studies suggest that multivesicular bodies are an intermediate stage in the formation of alpha-granules. In contrast, the kinetics and mode of appearance of dense granules during megakaryocytic maturation has remained poorly understood. Immunoelectron microscopy was used to monitor the appearance of dense granular markers (granulophysin and serotonin) on cryosections of human megakaryocytes (MKs) cultured from CD34(+) precursors. The monitoring was done on days 8 and 13 of culture. The data suggest that dense granules appear in immature MKs early during their maturation, concomitantly with alpha-granule formation. In MKs of intermediary maturation stage, granulophysin was mainly localized within dense granules and multivesicular bodies (MVBs), which were also labeled for serotonin. This study provides evidence that granulophysin is a dense granule marker in human MKs and that MVBs are an intermediary stage of dense granule maturation and probably constitute a sorting compartment between alpha-granules and dense granules. (Blood. 2000;95:4004-4007) PMID- 10845942 TI - Results of high-dose therapy for 1000 patients with multiple myeloma: durable complete remissions and superior survival in the absence of chromosome 13 abnormalities. AB - High-dose therapy (HDT) has increased complete remission (CR) rates and survival in multiple myeloma (MM). We now report on continuous CR (CCR) and associated prognostic factors in 1000 consecutive patients receiving melphalan-based tandem HDT. Five-year CCR was 52% among 112 CR patients without chromosome 13 (triangle up13) abnormalities and with beta-2-microglobulin 60% of neurons tested (11 of 18). Static (10- to 30-s) MC significantly (P < 0.05) increased discharge in four cells, two of which were also responsive to rhythmic MC. Rhythmic and sustained muscle stretch increased discharge (P < 0.05) in three of eight neurons tested. These data indicate that nucleus tractus solitarius neurons receive input from low- and high-threshold cutaneous mechanoreceptors, respond to capsaicin delivered into the hindlimb arterial supply, lack thermal sensitivity, and respond to activation of mechanosensitive as well as metabosensitive endings in skeletal muscle. PMID- 10846020 TI - Effects of swimming exercise on red blood cell rheology in trained and untrained rats. AB - Red blood cell (RBC) mechanical properties were investigated after swimming exercise in trained and untrained rats. A group of rats was trained for 6 wk (60 min swimming, daily), and another group was kept sedentary. Blood samples were obtained either within 5 min or 24 h after 60 min swimming in both groups. In the untrained rats, the RBC aggregation index decreased to 2.60 +/- 0.4 immediately after exercise from a control value of 6.73 +/- 0.18 (P < 0.01), whereas it increased to 13.13 +/- 0.66 after 24 h (P < 0.01). RBC transit time through 5 microm pores increased to 3.53 +/- 0.16 ms within 5 min after the exercise from a control value of 2.19 +/- 0. 07 ms (P < 0.005). A very significant enhancement (166%) in RBC lipid peroxidation was detected only after 24 h. In the trained group, the alterations in all these parameters were attenuated; there was a slight, transient impairment in RBC deformability (transit time = 2.64 +/- 0.13 ms), and lipid peroxidation was found to be unchanged. These findings suggest that training can significantly limit the hemorheological alterations related to a given bout of exercise. Whether this effect is secondary to the training induced reduction in the degree of metabolic and/or hormonal perturbation remains to be determined. PMID- 10846021 TI - Thoracoabdominal asynchrony failed to grade airway obstructions in foals. AB - Respiratory inductive plethysmography (RIP) can be used to obtain a valid measure of tidal volume in humans. This device also compares the contributions to ventilation of the thorax and abdomen. Although thoracoabdominal asynchrony is a prominent clinical feature for patients with airway obstruction, the accuracy of the RIP device to assess the severity of obstruction is unclear. This study analyzes how well RIP variables reflect the degree of a fixed external inspiratory plus expiratory resistive load in foals. Foals were employed because the species and age group are commonly afflicted with respiratory disease. Eight conscious, sedated (xylazine 1.25 mg/kg body wt) foals were subjected to randomly ordered resistive loads at the airway opening and, on a separate day, to histamine aerosol challenge. During resistive loading, phase angle changed significantly, as did phase relation (P < or = 0.05). However, no significant correlation was found between the degree of change in resistive load and the degree to which phase angle or relation was altered (r(s) = 0.41 and 0.25, respectively). In addition, neither phase angle nor relation changed significantly with histamine challenge. We conclude that, although RIP variables changed markedly with fixed upper airway resistive loading, the degree to which they changed was erratic and therefore not useful for grading these obstructions. Furthermore, RIP variables were insensitive measures of histamine-induced bronchoconstriction. PMID- 10846022 TI - Force-frequency relationship and potentiation in mammalian skeletal muscle. AB - Repetitive activation of a skeletal muscle results in potentiation of the twitch contractile response. Incompletely fused tetanic contractions similar to those evoked by voluntary activation may also be potentiated by prior activity. We aimed to investigate the role of stimulation frequency on the enhancement of unfused isometric contractions in rat medial gastrocnemius muscles in situ. Muscles set at optimal length were stimulated via the sciatic nerve with 50 micros duration supramaximal pulses. Trials consisted of 8 s of repetitive trains [5 pulses (quintuplets) 2 times per second or 2 pulses (doublets) 5 times per second] at 20, 40, 50, 60, 70, and 80 Hz. These stimulation frequencies represent a range over which voluntary activation would be expected to occur. When the frequency of stimulation was 20, 50, or 70 Hz, the peak active force (highest tension during a contraction - rest tension) of doublet contractions increased from 2.2 +/- 0.2, 4.1 +/- 0.4, and 4.3 +/- 0.5 to 3.1 +/- 0.3, 5.6 +/- 0.4, and 6.1 +/- 0.7 N, respectively. Corresponding measurements for quintuplet contractions increased from 2.2 +/- 0.2, 6.1 +/- 0.5, and 8.7 +/- 0.7 to 3.2 +/- 0.3, 7.3 +/- 0.6, and 9.0 +/- 0.7 N, respectively. Initial peak active force values were 27 +/- 1 and 61.5 +/- 5% of the maximal (tetanic) force for doublet and quintuplet contractions, respectively, at 80 Hz. With doublets, peak active force increased at all stimulation frequencies. With quintuplets, peak active force increased significantly for frequencies up to 60 Hz. Twitch enhancement at the end of the 8 s of repetitive stimulation was the same regardless of the pattern of stimulation during the 8 s, and twitch peak active force returned to prestimulation values by 5 min. These experiments confirm that activity-dependent potentiation is evident during repeated, incompletely fused tetanic contractions over a broad range of frequencies. This observation suggests that, during voluntary motor unit recruitment, derecruitment or decreased firing frequency would be necessary to achieve a fixed (submaximal) target force during repeated isometric contractions over this time period. PMID- 10846023 TI - Effects of resistance exercise combined with moderate vascular occlusion on muscular function in humans. AB - Acute and long-term effects of resistance exercise combined with vascular occlusion on muscular function were investigated. Changes in integrated electromyogram with respect to time (iEMG), vascular resistive index, and plasma lactate concentration were measured in five men either during or after elbow flexion exercises with the proximal end of the arm occluded at 0-100 mmHg. The mean iEMG, postexercise hyperemia, and plasma lactate concentration were all elevated with the increase in occlusion pressure at a low-intensity exercise, whereas they were unchanged with the increase in occlusion pressure at high intensity exercise. To investigate the long-term effects of low-intensity exercise with occlusion, older women (n = 24) were subjected to a 16-wk exercise training for elbow flexor muscles, in which low-intensity [ approximately 50-30% one repetition maximum (1 RM)] exercise with occlusion at approximately 110 mmHg (LIO), low-intensity exercise without occlusion (LI), and high- to medium intensity ( approximately 80-50% 1 RM) exercise without occlusion (HI) were performed. Percent increases in both cross-sectional area and isokinetic strength of elbow flexor muscles after LIO were larger than those after LI (P < 0.05) and similar to those after HI. The results suggest that resistance exercise at an intensity even lower than 50% 1 RM is effective in inducing muscular hypertrophy and concomitant increase in strength when combined with vascular occlusion. PMID- 10846024 TI - Cerebral and intestinal perfusion and metabolism in normocythemic hyperviscous hypoxic newborn pigs. AB - We studied the effects of hypoxia on cerebral cortical and intestinal perfusion and metabolism in normocythemic hyperviscous newborn pigs. Seven pigs were made hyperviscous by an injection of cryoprecipitate, increasing viscosity from 5.8 +/ 0.9 to 9.0 +/- 1. 2 (SD) cycles/s. Six normoviscous pigs received 0.9% NaCl. Reducing the inspired O(2) decreased the arterial O(2) content (Ca(O(2))) from 9.5 +/- 1.6 to 3.6 +/- 1.3 ml O(2)/100 ml. Increases in brain and decreases in gastrointestinal blood flow at the lower Ca(O(2)) values were similar between the groups. During hypoxia, blood flow to stomach, distal intestinal mucosa, and large intestines was lower (-50, -23, and -28%, respectively) in the hyperviscous than normoviscous group. At the lower Ca(O(2)) values, cerebral cortical vascular resistance decreased in both groups and intestinal vascular resistance increased (+257%) in the hyperviscous but not in the normoviscous group. During hypoxia, systemic oxygen delivery decreased, extraction increased, and uptake did not change; cerebral cortical O(2) delivery, extraction, and uptake did not change; and intestinal O(2) delivery decreased, extraction increased, and uptake did not change in both groups. Our study demonstrated that 1) during hypoxia, increases in systemic O(2) extraction compensated for decreases in delivery and systemic uptake did not change; vasodilation sustained cerebral cortical O(2) delivery and preserved metabolism; increases in intestinal oxygen extraction offset decreases in delivery and uptake was preserved; and 2) nonpolycythemic hyperviscosity did not have a major influence on cardiovascular or metabolic responses to hypoxia, except for modest effects on intestinal resistance and perfusion to certain gastrointestinal regions. We conclude that, under normocythemic conditions, a moderate increase in viscosity does not have a major impact on hemodynamic or metabolic adjustments to hypoxia in newborn pigs. PMID- 10846025 TI - Effects of exercise on insulin distribution and action in testosterone-treated oophorectomized female rats. AB - Administration of testosterone (T) to oophorectomized (Ovx) female rats is followed by severe insulin resistance, localized to postreceptor cellular events in the muscle. In this study, intervention by exercise was introduced to examine whether circulatory adaptations are involved in insulin resistance. Two groups of Ovx rats were studied: one group was given T (Ovx+T); another group had free access to running wheels (Ovx+T+Ex). In addition, one control group (sham operated) was studied. Insulin sensitivity was measured with the euglycemic hyperinsulinemic clamp technique (submaximal) for 150 min. Muscle interstitial glucose and insulin concentrations were measured by microdialysis. The measurements showed that, in Ovx+T rats, the onset of insulin action was significantly (P < 0.05) slower during the first 95 min of the clamp compared with that in Ovx+T+Ex and controls. Muscle interstitial concentrations of insulin but not glucose were lower in both Ovx+T and Ovx+T+Ex rats than in controls throughout the clamp. It was concluded that physical exercise prevented the slow onset of insulin action in Ovx+T rats without changing the distribution time of muscle interstitial insulin. The results indicate that hyperandrogenicity is characterized by delayed muscle insulin action. Physical exercise reverses these defects without any beneficial effect on muscle interstitial insulin concentrations. PMID- 10846026 TI - Fatiguing contractions of tongue protrudor and retractor muscles: influence of systemic hypoxia. AB - The influence of systemic hypoxia on the endurance performance of tongue protrudor and retractor muscles was examined in anesthetized, ventilated rats. Tongue protrudor (genioglossus) or retractor (hyoglossus and styloglossus) muscles were activated via medial or lateral XII nerve branch stimulation (0.1-ms pulse; 40 Hz; 330-ms trains; 1 train/s). Maximal evoked potentials (M waves) of genioglossus and hyoglossus were monitored with electromyography. Fatigue tests were performed under normoxic and hypoxic (arterial PO(2) = 50 +/- 1 Torr) conditions in separate animals. The fatigue index (FI; %initial force) after 5 min of normoxic stimulation was 85 +/- 6 and 79 +/- 7% for tongue protrudor and retractor muscles, respectively; these values were significantly lower during hypoxia (protrudor FI = 52 +/- 10, retractor FI = 18 +/- 6%; P < 0.05). Protrudor and retractor muscle M-wave amplitude declined over the course of the hypoxic fatigue test but did not change during normoxia (P < 0.05). We conclude that hypoxia attenuates tongue protrudor and retractor muscle endurance performance; potential mechanisms include neuromuscular transmission failure and/or diminished sarcolemmal excitability. PMID- 10846027 TI - Postactivation potentiation, fiber type, and twitch contraction time in human knee extensor muscles. AB - In small mammals, muscles with shorter twitch contraction times and a predominance of fast-twitch, type II fibers exhibit greater posttetanic twitch force potentiation than muscles with longer twitch contraction times and a predominance of slow-twitch, type I fibers. In humans, the correlation between potentiation and fiber-type distribution has not been found consistently. In the present study, postactivation potentiation (PAP) was induced in the knee extensors of 20 young men by a 10-s maximum voluntary isometric contraction (MVC). Maximal twitch contractions of the knee extensors were evoked before and after the MVC. A negative correlation (r = -0. 73, P < 0.001) was found between PAP and pre-MVC twitch time to peak torque (TPT). The four men with the highest (HPAP, 104 +/- 11%) and lowest (LPAP, 43 +/- 7%) PAP values (P < 0.0001) underwent needle biopsies of vastus lateralis. HPAP had a greater percentage of type II fibers (72 +/- 9 vs. 39 +/- 7%, P < 0.001) and shorter pre-MVC twitch TPT (61 +/- 12 vs. 86 +/- 7 ms, P < 0.05) than LPAP. These data indicate that, similar to the muscles of small mammals, human muscles with shorter twitch contraction times and a higher percentage of type II fibers exhibit greater PAP. PMID- 10846028 TI - Near-maximal fractional oxygen extraction by active skeletal muscle in patients with chronic heart failure. AB - Systemic oxygen uptake and deep femoral vein oxygen content were determined at peak exercise in 53 patients with chronic heart failure with impaired systolic function (mean left ventricular ejection fraction 0.18; n = 41) or preserved systolic function (mean left ventricular ejection fraction 0.70; n = 12) and in 6 age-matched sedentary normal subjects. At peak exercise, deep femoral vein oxygen content in heart failure patients with impaired systolic function and preserved systolic function were similar, both significantly lower than that of normal subjects (2.5 +/- 0.1, 2.9 +/- 0.2, and 5.0 +/- 0.1 ml/100 ml, respectively; P < 0.05). Deep femoral venous oxygen content was lower in patients with the greater impairment of aerobic capacity, regardless of the underlying systolic function (r = 0.72, P < 0.01). Fractional oxygen extraction in the skeletal muscle at peak exercise is enhanced in patients with chronic heart failure when compared with normal subjects, in proportion to the degree of aerobic impairment. PMID- 10846029 TI - Analysis of the mechanisms of expiratory asynchrony in pressure support ventilation: a mathematical approach. AB - A mathematical model was developed to analyze the mechanisms of expiratory asynchrony during pressure support ventilation (PSV). Solving the model revealed several results. 1) Ratio of the flow at the end of patient neural inspiration to peak inspiratory flow (VTI/V(peak)) during PSV is determined by the ratio of time constant of the respiratory system (tau) to patient neural inspiratory time (TI) and the ratio of the set pressure support (Pps) level to maximal inspiratory muscle pressure (Pmus max). 2) VTI/V(peak) is affected more by tau/TI than by Pps/Pmus max. VTI/V(peak) increases in a sigmoidal relationship to tau/TI. An increase in Pps/Pmus max slightly shifts the VTI/V(peak)-tau/TI curve to the right, i.e., VTI/V(peak) becomes lower as Pps/Pmus max increases at the same tau/TI. 3) Under the selected adult respiratory mechanics, VTI/V(peak) ranges from 1 to 85% and has an excellent linear correlation with tau/TI. 4) In mechanical ventilators, single fixed levels of the flow termination criterion will always have chances of both synchronized termination and asynchronized termination, depending on patient mechanics. An increase in tau/TI causes more delayed and less premature termination opportunities. An increase in Pps/Pmus max narrows the synchronized zone, making inspiratory termination predisposed to be in asynchrony. Increasing the expiratory trigger sensitivity of a ventilator shifts the synchronized zone to the right, causing less delayed and more premature termination. Automation of expiratory trigger sensitivity in future mechanical ventilators may also be possible. In conclusion, our model provides a useful tool to analyze the mechanisms of expiratory asynchrony in PSV. PMID- 10846030 TI - Dietary carbohydrate, muscle glycogen content, and endurance performance in well trained women. AB - This study examined the ability of well-trained eumenorrheic women to increase muscle glycogen content and endurance performance in response to a high carbohydrate diet (HCD; approximately 78% carbohydrate) compared with a moderate carbohydrate diet (MD; approximately 48% carbohydrate) when tested during the luteal phase of the menstrual cycle. Six women cycled to exhaustion at approximately 80% maximal oxygen uptake (VO(2 max)) after each of the randomly assigned diet and exercise-tapering regimens. A biopsy was taken from the vastus lateralis before and after exercise in each trial. Preexercise muscle glycogen content was high after the MD (625.2 +/- 50.1 mmol/kg dry muscle) and 13% greater after the HCD (709.0 +/- 44.8 mmol/kg dry muscle). Postexercise muscle glycogen was low after both trials (MD, 91.4 +/- 34.5; HCD, 80.3 +/- 19.5 mmol/kg dry muscle), and net glycogen utilization during exercise was greater after the HCD. The subjects also cycled longer at approximately 80% VO(2 max) after the HCD vs. MD (115:31 +/- 10:47 vs. 106:35 +/- 8:36 min:s, respectively). In conclusion, aerobically trained women increased muscle glycogen content in response to a high dietary carbohydrate intake during the luteal phase of the menstrual cycle, but the magnitude was smaller than previously observed in men. The increase in muscle glycogen, and possibly liver glycogen, after the HCD was associated with increased cycling performance to volitional exhaustion at approximately 80% VO(2 max). PMID- 10846031 TI - Linkage of hiccup with heartbeat. AB - We explored a possible link between the cardiac cycle and the timing of recurrent hiccups in 10 patients with chronic, intractable hiccups. Recordings made during daytime naps in a sleep laboratory included sleep state; electrocardiogram; and respiration by means of a thermistor to detect airflow, bands around the rib cage and abdomen to assess expansion, and a bipolar surface electrode electromyogram over parasternal intercostal muscles. Hiccups could be detected on the abdominal bands and the parasternal electromyogram. The time of occurrence of each hiccup and each R wave in a continuous tracing of 100 or more hiccups were recorded and analyzed together with semiquantitive estimates of the phase of hiccup respiration. Whereas the hiccup rate ranged from approximately one-third to one eighth of heart rate and was more variable than heart rate, hiccups showed a tendency, stronger in some subjects than others, to occur in midsystole. Variation in R-wave-R-wave (R-R) interval in association with hiccups was found in five patients. In three of these patients, hiccups were synchronized with respiration so that the cyclic change in R-R interval posthiccup could be explained as sinus arrhythmia, but, in two patients, the hiccups were not synchronized with respiration, so that hiccups are most likely responsible for the variation in heart rate. Also, the variation of R-R interval with hiccups suggests that there is some phasic autonomic efferent activity associated with hiccups. PMID- 10846032 TI - Effects of activation frequency on dynamic performance of human fresh and fatigued muscles. AB - The force-frequency relationship for an individual muscle depends on the fatigue state, the length at which it is activated, and the muscle's activation history. The relationship among stimulation frequency and dynamic (nonisometric) muscle performance measurements (e.g., excursion, work, peak power, and average power) has not been reported. The purpose of this study was to identify the relationship between stimulation frequency and dynamic performance measurements for fresh and fatigued muscles. Constant-frequency and catchlike-inducing trains (CFT and CIT, respectively) were tested. When fresh, interpulse intervals of 40-50 ms [20-25 pulses/s (pps)] produced maximum performance for CFTs. For CITs, maximum performance occurred at interpulse intervals of 50-60 ms ( approximately 16-20 pps). Generally, CFTs produced slightly greater performance than did CITs. When fatigued, however, CITs produced greater performance than did CFTs. Maximum performance for CFTs occurred at interpulse intervals of 20-40 ms (25-50 pps) and at 30-50 ms (20-33 pps) for CITs. Enhancement of performance by CITs when fatigued may be due to less susceptibility to impairments in excitation contraction coupling and greater ability to maintain rates of rise of force than CFTs. PMID- 10846033 TI - Treadmill exercise training blunts suppression of splenic natural killer cell cytolysis after footshock. AB - This study extended to treadmill exercise training our prior report (Dishman RK, Warren JM, Youngstedt SD, Yoo H, Bunnell BN, Mougey EH, Meyerhoff JL, Jaso Friedmann L, and Evans DL. J Appl Physiol 78: 1547-1554, 1995) that activity wheel running abolished the suppression of footshock-induced natural killer (NK) cell cytolysis. Twenty-four male Fischer 344 rats were assigned to one of three groups (n = 8, all groups): 1) a home-cage control group, 2) a sedentary treatment group, or 3) a treadmill-running group (0 degrees incline, 25 m/min, 35 min/day, 6 days/wk). After 6 wk, the treadmill and sedentary groups received 2 days of footshock. Splenic NK cytotoxicity was determined by standard 4-h (51)Cr release assay. Percentages of lymphocytes were determined by flow cytometry. Plasma levels of ACTH, corticosterone, and prolactin concentration were measured by radioimmunoassay. After footshock, percentage of lysis relative to home-cage controls was 40% and 80% for sedentary and treadmill-trained animals, respectively (P < 0.05). Our results indicate that the protective effect of chronic exercise on innate cellular immunity in the Fischer 344 male rat is not restricted to activity wheel running, nor is it explained by elevations in basal NK activity, increased percentages of splenic NK and cytotoxic T cells, or increased plasma levels of ACTH, corticosterone, and prolactin. PMID- 10846034 TI - High-impact exercise and growing bone: relation between high strain rates and enhanced bone formation. AB - We investigated whether high-impact drop jumps could increase bone formation in the middiaphyseal tarsometatarsus of growing rooster. Roosters were designated as sedentary controls (n = 10) or jumpers (n = 10). Jumpers performed 200 drop jumps per day for 3 wk. The mechanical milieu of the tarsometatarsus was quantified via in vivo strain gauges. Indexes of bone formation and mechanical parameters were determined in each of twelve 30 degrees sectors subdividing the middiaphyseal cortex. Compared with baseline walking, drop jumping produced large peak strain rates (+740%) in the presence of moderately increased peak strain magnitudes (+30%) and unaltered strain distributions. Bone formation rates were significantly increased by jump training at periosteal (+40%) and endocortical surfaces (+370%). Strain rate was significantly correlated with the specific sites of increased formation rates at endocortical but not at periosteal surfaces. Previously, treadmill running did not enhance bone growth in this model. Comparing the mechanical milieus produced by running and drop jumps revealed that jumping significantly elevated only peak strain rates. This further emphasized the sensitivity of immature bone to high strain rates. PMID- 10846035 TI - Basal lung mechanics and airway and pulmonary vascular responsiveness in different inbred mouse strains. AB - Little is known about interstrain variations in baseline lung functions or smooth muscle contractility in murine lungs. We therefore examined basal lung mechanics and airway, as well as vascular reactivity to methacholine, thromboxane (using U 46619), and endothelin-1 (ET-1), A/J, AKR, BALB/c, C3H/HeN, C57BL/6, and SCID mice. All experiments were performed with isolated perfused mouse lungs. Except AKR mice (which were excluded from further analysis), all other strains showed stable pulmonary compliance, pulmonary resistance, and pulmonary arterial pressure within a control period of 45 min. Among these strains, C3H/HeN mice exhibited higher dynamic pulmonary compliance and lower pulmonary resistance, whereas SCID mice had higher baseline pulmonary resistance than the other strains. Concentration-response experiments with methacholine showed a lower airway reactivity for C57BL/6 mice compared with the other strains. Perfusion with 1 microM U-46619 or 100 nM ET-1 revealed a similar pattern: the agonist inducible broncho- and vasoconstriction was lower in C57BL/6 mice than in all other strains, whereas it tended to be higher in SCID mice. The present study demonstrates a correlation between airway and vascular responsiveness in all tested strains. SCID mice are hyperreactive, whereas C57BL/6 mice are hyporeactive, to smooth muscle constrictors. Lung mechanics, as well as airway and vascular responsiveness, appear to be genetically controlled. PMID- 10846036 TI - Effects of exercise and n-3 fatty acids on postprandial lipemia. AB - Because n-3 fatty acid ingestion and aerobic exercise each has been associated with diminished postprandial lipemia (PPL), the purpose of this study was to evaluate the effect of a combination of these two factors on PPL. Sedentary men underwent a standard dietary preparation, including a 12-h fast before each trial. Six subjects performed a control trial (fat meal, 100 g fat) and an n-3 fatty acid trial (fat meal after 3 wk of n-3 fatty acid supplementation at 4 g/day). In a parallel experiment, six different subjects underwent a control trial and n-3 fatty acid supplementation + 60 min of exercise before ingestion of the fat meal. Supplementation with n-3 fatty acid significantly decreased baseline triglyceride (TG) concentrations but did not significantly affect PPL. The combination of n-3 fatty acid and exercise had no effect on the postprandial TG response. The present study suggests that n-3 fatty acid supplementation lowers resting TG concentrations but inhibits the beneficial effect of aerobic exercise on the postprandial TG response. PMID- 10846037 TI - Evaluation of the cerebral hemodynamic response to rhythmic handgrip. AB - The response of the cerebral circulation to exercise has been studied with transcranial Doppler ultrasound (TCD) because this modality provides continuous measurements of blood velocity and is well suited for the exercise environment. The use of TCD as an index of cerebral blood flow, however, requires the assumption that the diameter of the insonated vessel is constant. Here, we examine this assumption for rhythmic handgrip using a spectral index designed to measure trends in vessel flow. Nineteen normal subjects were studied during 5 min of volitional maximum rhythmic right handgrip at 1 Hz. TCD velocities from both middle arteries (left and right), blood pressure, and end-tidal PCO(2) were recorded every 10 s. A spectral weighted sum was also calculated as a flow index (FI). Averages were computed from the last 2 min of handgrip. Relative changes in velocity, FI, and pressure were calculated. The validity of FI was tested by comparing the change in diameter derived from equations relating flow and diameter. Mean blood pressure increased 23.8 +/- 17.8% (SD), and velocity increased 13.3 +/- 9.8% (left) and 9.6 +/- 8.3% (right). Although the mean change in FI was small [2.0 +/- 18. 2% (left) and 4.7 +/- 29.7% (right)], the variation was high: some subjects showed a significant increase in FI and others a significant decrease. Diameter estimates from two equations relating flow and luminal area were not significantly different. Decreases in FI were associated with estimated diameter decreases of 10%. Our data suggest that the cerebral blood flow (CBF) response to rhythmic handgrip is heterogeneous and that middle cerebral artery flow can decrease in some subjects, in agreement with prior studies using the Kety-Schmidt technique. We speculate that the velocity increase is due to sympathetically mediated vasoconstriction rather than a ubiquitous flow increase. Our data suggest that the use of ordinary TCD velocities to interpret the CBF response during exercise may be invalid. PMID- 10846038 TI - EP(2) receptor mediates bronchodilation by PGE(2) in mice. AB - PGE(2) is an important cyclooxygenase product that modulates airway inflammatory and smooth muscle responses. Signal transduction is mediated by four EP receptor subtypes that cause distinct effects on cell metabolism. To determine the role of EP(2) receptor activation, we produced a mouse lacking the EP(2) receptor by targeted gene disruption. The effect of aerosolized PGE(2) and other agonists was measured using barometric plethysmography and by measurements of lung resistance in mechanically ventilated mice. Inhalation of PGE(2) inhibited methacholine responses in wild-type but not in mice lacking the EP(2) receptor [EP(2)(-/-)]. After airway constriction was induced by methacholine aerosol, PGE(2) reduced the airway constriction enhanced pause in wild-type mice (from 0.88 +/- 0.15 to 0.55 +/- 0.06) but increased it in EP(2)(-/-) mice (from 0.73 +/- 0. 08 to 1.27 +/- 0.19). Similar results were obtained in mechanically ventilated mice. These data indicate that the EP(2) receptor mediates the bronchodilation effect of PGE(2). PMID- 10846040 TI - Aortic input impedance in infants and children. AB - Flow and pressure measurements were performed in the ascending aortas of six pediatric patients ranging in age from 1 to 4 yr and in weight from 7.2 to 16.4 kg. From these measurements, input impedance was calculated. It was found that total vascular resistance decreased with increasing patient weight and was approximately one to three times higher than those of adults. Conductance per unit weight was relatively constant but was approximately three times higher than for adults. Strong inertial character was observed in the impedance of four of the six patients. Among a three-element and two four-element lumped-parameter models, the model with characteristic aortic resistor (R(c)) and inertance in series followed by parallel peripheral resistor (R(p)) and compliance fitted the data best. R(p) decreased with increasing patient weight and was one to three times higher than in adults, and R(c) decreased with increasing patient weight and was 2 to 15 times higher. The R(p)-to-R(c) ratio differed significantly between infants and children vs. adults. The results suggested that R(p) developed more rapidly with patient weight than did R(c). Compliance values increased with increasing patient weight and were 3 to 16 times lower than adult values. PMID- 10846039 TI - Activation of AMP-activated protein kinase increases mitochondrial enzymes in skeletal muscle. AB - Muscle contraction causes an increase in activity of 5'-AMP-activated protein kinase (AMPK). This study was designed to determine whether chronic chemical activation of AMPK will increase mitochondrial enzymes, GLUT-4, and hexokinase in different types of skeletal muscle of resting rats. In acute studies, rats were subcutaneously injected with either 5-aminoimidazole-4-carboxamide-1-beta-D ribofuranoside (AICAR; 1 mg/g body wt) in 0.9% NaCl or with 0.9% NaCl alone and were then anesthetized for collection and freezing of tissues. AMPK activity increased in the superficial, white region of the quadriceps and in soleus muscles but not in the deep, red region of the quadriceps muscle. Acetyl-CoA carboxylase (ACC) activity, a target for AMPK, decreased in all three muscle types in response to AICAR injection but was lowest in the white quadriceps. In rats given daily, 1 mg/g body wt, subcutaneous injections of AICAR for 4 wk, activities of citrate synthase, succinate dehydrogenase, and malate dehydrogenase were increased in white quadriceps and soleus but not in red quadriceps. Cytochrome c and delta-aminolevulinic acid synthase levels were increased in white, but not red, quadriceps. Carnitine palmitoyl-transferase and hydroxy-acyl CoA dehydrogenase were not significantly increased. Hexokinase was markedly increased in all three muscles, and GLUT-4 was increased in red and white quadriceps. These results suggest that chronic AMPK activation may mediate the effects of muscle contraction on some, but not all, biochemical adaptations of muscle to endurance exercise training. PMID- 10846042 TI - Effects of length oscillation on the subsequent force development in swine tracheal smooth muscle. AB - It has been shown that deep inspiration (DI) taken before application of bronchoconstricting stimuli causes a reduction in the subsequent bronchoconstriction; a fast DI has a greater inhibitory effect than a slow DI. We hypothesize that periodic length changes imposed on a relaxed airway smooth muscle (ASM) would attenuate subsequent bronchoconstriction by disrupting the organization of the contractile apparatus, and this could be an important mechanism for the observed bronchoprotective effect of DI and tidal breathing. Length oscillations of different amplitude, frequency, and duration were applied to a relaxed muscle. The effects of such perturbations on force development were then assessed. Results show that oscillations reduce the subsequent force generation and that the magnitude of force reduction is proportional to amplitude and duration of the length oscillation. After the oscillation, isometric force recovered to the preoscillation level in a series of isometric contractions, and the rate of recovery was facilitated by frequent stimulation. The in vitro behavior of ASM found in this study could account for the observed temporary reduction in bronchoconstriction subsequent to a DI. PMID- 10846041 TI - Transient enhancement of GLUT-4 levels in rat epitrochlearis muscle after exercise training. AB - The purpose of the present study was to examine the effect of detraining on the glucose transport system after short-term swim training (5 days), long-term swim training (5 wk), and treadmill run training (5 wk). Skeletal muscles were isolated from female Wistar rats at 24 or 48 h posttraining. SST produces a 48% increase in GLUT-4 mRNA, a 30% increase in GLUT-4 protein, and a 60% increase in insulin-stimulated glucose transport activity at 24 h posttraining but not at 48 h posttraining. Similar to SST, long-term swim training produces a 60% increase in GLUT-4 mRNA and a 30% increase in GLUT-4 protein content at 24 h posttraining but not at 48 h posttraining. Finally, treadmill run training produces a transient 35% increase in GLUT-4 protein content that is completely reversed at 48 h after the last bout of exercise. These results demonstrate that the increase in GLUT-4 mRNA and GLUT-4 protein occurs during the first week of exercise training and is rapidly lost after training cessation. We believe that the transient enhancement in GLUT-4 protein after exercise training is due to a short GLUT-4 half-life, a process that is primarily regulated by pretranslational mechanisms. PMID- 10846043 TI - Regional body composition changes in women after 6 months of periodized physical training. AB - Data are lacking regarding regional morphological changes among women after prolonged physical training. This study employed dual-energy X-ray absorptiometry to assess changes in whole body and regional (i.e., trunk, legs, arms) fat mass, lean mass, and bone mineral content body composition adaptations in 31 healthy women pre-, mid-, and post-6 mo of periodized physical training. These results were compared with those of 1) a control group of women who had not undergone the training program and were assessed pre- and post-6 mo and 2) a group of 18 men that was tested only once. Additionally, magnetic resonance imaging was used to assess changes in muscle morphology of the thigh in a subset of 11 members of the training group. Physical training consisted of a combination of aerobic and resistance exercise in which the subjects engaged for 5 days/wk for 24 wk. Overall, the training group experienced a 2.2% decrease, a 10% decrease, and a 2.2% increase for body mass, fat mass, and soft tissue lean mass, respectively. No changes in bone mineral content were detected. The women had less of their soft tissue lean mass distributed in their arms than did the men, both before and after the women were trained. Novel to this study were the striking differences in the responses in the tissue composition of the arms (31% loss in fat mass but no change in lean mass) compared with the legs (5.5% gain in lean mass but no change in fat mass). There was a 12% fat loss in the trunk with no change in soft tissue lean mass. Dual-energy X-ray absorptiometry and magnetic resonance imaging fat mass measurements showed good agreement (r = 0. 72-0.92); their lean mass measurements were similar as well, showing approximately 5.5% increases in leg lean tissue. These findings show the importance of considering regional body composition changes, rather than whole body changes alone when assessing the effects of a periodized physical training program. PMID- 10846044 TI - A three-dimensional model of the human pulmonary acinus. AB - A three-dimensional (3-D) model of the human pulmonary acinus, a gas exchange unit, is constructed with a labyrinthine algorithm generating branching ducts that fill a given space completely. Branching down to the third respiratory bronchioles is generated with the proposed algorithm. A subacinus, a region supplied by the last respiratory bronchiole, is approximated to be a set of cubic cells with a side dimension of 0.5 mm. The labyrinthine algorithm is used to determine a pathway through all cells only once, except at branching points with the smallest path lengths. In choosing each step of a pathway, random variables are used. Resulting labyrinths have equal mean path lengths and equal surface areas of inner walls. An alveolus can be generated by attaching alveolar septa, 0.25 mm long and 0.1 mm wide, to the inner walls. Total alveolar surface area and numbers of alveolar ducts, alveolar sacs, and alveoli in our 3-D acinar model are in good accordance with those reported in the literature. PMID- 10846045 TI - Exercise alters fractal dimension and spatial correlation of pulmonary blood flow in the horse. AB - We determined the changes in fractal dimensions and spatial correlations of regional pulmonary blood flow with increasing exercise in race horses (n = 4) by using 15-microm fluorescent microspheres. Fluorescence was measured to quantitate regional blood to 1.3-cm(3) samples (n = 1,621-2,503). Perfusion distributions were characterized with fractal dimensions (a measure of spatial variability) and spatial correlations. On average, the fractal dimension decreased with exercise (trot 1.216 to gallop 1.173; P < 0. 05) despite a variable fractal dimension at rest. Spatial correlation of flow to neighboring pieces increased with exercise (trot 0.57 +/- 0.074 to gallop 0.73 +/- 0.051) and was inversely correlated with fractal dimension, indicating better spatial correlation as blood flow distribution becomes more uniform. This is the first study to document a change in fractal dimension as a result of increasing pulmonary blood flow. Spatial differences in response to vasoregulatory mediators may play a role in this phenomenon. PMID- 10846046 TI - Imaging obstructed ventilation with NMR using inert fluorinated gases. AB - We partially obstructed the left bronchi of rats and imaged an inert insoluble gas, SF(6), in the lungs with NMR using a technique that clearly differentiates obstructed and normal ventilation. When the inhaled fraction of O(2) is high, SF(6) concentrates dramatically in regions of the lung with low ventilation-to perfusion ratios (VA/Q); therefore, these regions are brighter in an image than where VA/Q values are normal or high. A second image, made when the inhaled fraction of O(2) is low, serves as a reference because the SF(6) fraction is nearly uniform, regardless of VA/Q. The quotient of the first and second images displays the low-VA/Q regions and is corrected for other causes of brightness variation. The technique may provide sufficient quantification of VA/Q to be a useful research tool. The noise in the quotient image is described by the probability density function for the quotient of two normal random variables. When the signal-to-noise ratio of the denominator image is >10, the signal-to noise ratio of the quotient image is similar to that of the parent images and decreases with pixel value. PMID- 10846047 TI - Oxygen sensing by the carotid body chemoreceptors. AB - Carotid bodies are sensory organs that detect changes in arterial blood oxygen, and the ensuing reflexes are critical for maintaining homeostasis during hypoxemia. During the past decade, tremendous progress has been made toward understanding the cellular mechanisms underlying oxygen sensing at the carotid body. The purpose of this minireview is to highlight some recent concepts on sensory transduction and transmission at the carotid body. A bulk of evidence suggests that glomus (type I) cells are the initial site of transduction and that they release transmitters in response to hypoxia, which causes depolarization of nearby afferent nerve endings, leading to an increase in sensory discharge. There are two main hypotheses to explain the transduction process that triggers transmitter release. One hypothesis assumes that a biochemical event associated with a heme protein triggers the transduction cascade. The other hypothesis suggests that a K(+) channel protein is the oxygen sensor and that inhibition of this channel by hypoxia leading to depolarization is a seminal event in transduction. Although there is body of evidence supporting and questioning each of these, this review will try to point out that the truth lies somewhere in an interrelation between the two. Several transmitters have been identified in glomus cells, and they are released in response to hypoxia. However, their precise roles in sensory transmission remain uncertain. It is hoped that future studies involving transgenic animals with targeted disruption of genes encoding transmitters and their receptors may resolve some of the key issues surrounding the sensory transmission at the carotid body. Further studies are necessary to identify whether a single sensor or multiple oxygen sensors are needed for the transduction process. PMID- 10846048 TI - Developmental aspects of oxygen sensing by the carotid body. AB - The carotid body chemoreceptors, the major hypoxia sensory organs for the respiratory system, undergo a significant increase in their hypoxia responsiveness in the postnatal period. This is manifest by a higher level of afferent nerve activity for a given level of arterial oxygen tension. The mechanism for the enhanced sensitivity is unresolved, but most work has focused on the glomus cell, a secretory cell apposed to the afferent nerve ending and believed to be the site of hypoxia transduction. The glomus cell secretory response to hypoxia increases postnatally, and this is correlated with an enhanced calcium rise in response to hypoxia and an increase in oxygen-sensitive potassium currents. These changes are sensitive to the level of hypoxia in the postnatal period, and significant impairment of organ function is observed with postnatal hypoxia as well as postnatal hyperoxia. Although many questions remain, especially with regard to the coupling of glomus cells to nerve endings, the use of cellular and molecular techniques should offer resolution in the near future. PMID- 10846049 TI - Pulmonary expression of early growth response-1: biphasic time course and effect of oxygen concentration. AB - Hypoxia induces complex adaptive responses. In this report, induction of early growth response-1 (Egr-1) transcripts in lungs of mice subjected to hypoxia is shown to be dose and time dependent. Within 30 min of hypoxia, Egr-1 transcripts were approximately 20-fold elevated in 6% oxygen, approximately 5.2-fold increased by 10% oxygen, and returned to the normoxic baseline by 12% oxygen. Time course studies up to 48 h showed a biphasic profile with an initial steep rise in Egr-1 transcripts after 0.5 h of hypoxia and a second elevation beginning after 20-24 h. Hypoxic induction of Egr-1 was paralleled by enhanced expression of the downstream target gene tissue factor. Egr-1 and tissue factor antigen were visualized in bronchial and vascular smooth muscle and in alveolar macrophages. Egr-1 has the capacity to modulate expression of genes involved in the remodeling of the extracellular matrix and properties of smooth muscle, thus possibly contributing to the pulmonary response to chronic hypoxia. PMID- 10846050 TI - Genetic determinants of acute hypoxic ventilation: patterns of inheritance in mice. AB - Acutely lowering ambient O(2) tension increases ventilation in many mammalian species, including humans and mice. Inheritance patterns among kinships and between mouse strains suggest that a robust genetic influence determines individual hypoxic ventilatory responses (HVR). Here, we tested specific genetic hypotheses to describe the inheritance patterns of HVR phenotypes among two inbred mouse strains and their segregant and nonsegregant progeny. Using whole body plethysmography, we assessed the magnitude and pattern of ventilation in C3H/HeJ (C3) and C57BL/6J (B6) progenitor strains at baseline and during acute (3 5 min) hypoxic [mild hypercapnic hypoxia, inspired O(2) fraction (FI(O(2))) = 0.10] and normoxic (mild hypercapnic normoxia, FI(O(2)) = 0.21) inspirate challenges in mild hypercapnia (inspired CO(2) fraction = 0.03). First- and second-filial generations and two backcross progeny were also studied to assess response distributions of HVR phenotypes relative to the parental strains. Although the minute ventilation (VE) during hypoxia was comparable between the parental strains, breathing frequency (f) and tidal volume were significantly different; C3 mice demonstrated a slow, deep HVR relative to a rapid, shallow phenotype of B6 mice. The HVR profile in B6C3F(1)/J mice suggested that this offspring class represented a third phenotype, distinguishable from the parental strains. The distribution of HVR among backcross and intercross offspring suggested that the inheritance patterns for f and VE during mild hypercapnic hypoxia are consistent with models that incorporate two genetic determinants. These results further suggest that the quantitative genetic expression of alleles derived from C3 and B6 parental strains interact to significantly attenuate individual HVR in the first- and second-filial generations. In conclusion, the genetic control of HVR in this model was shown to exhibit a relatively simple genetic basis in terms of respiratory timing characteristics. PMID- 10846051 TI - Hypoxic excitation in neurons cultured from the rostral ventrolateral medulla of the neonatal rat. AB - Neurons within cardiorespiratory regions of the rostral ventrolateral medulla (RVLM) have been shown to be excited by local hypoxia. To determine the electrophysiological properties of these excitatory responses to hypoxia, we developed a primary dissociated cell culture system to examine the intrinsic response of RVLM neurons to hypoxia. Neonatal rat neurons plated on medullary astrocyte monolayers were studied using the whole cell perforated patch-clamp technique. Sodium cyanide (NaCN, 0.5-10 mM) was used, and membrane potential (V(m)), firing frequency, and input resistance were examined. In 11 of 19 neurons, NaCN produced a V(m) depolarization, an increase in firing frequency, and a decrease in input resistance, suggesting the opening of a cation channel. The hypoxic depolarization had a linear dose response and was dependent on baseline V(m), with a greater response at more hyperpolarized V(m). In 8 of 19 neurons, NaCN produced a V(m) hyperpolarization, decrease in firing frequency, and variable changes in input resistance. The V(m) hyperpolarization exhibited an all-or-none dose response and was independent of baseline V(m). These differential responses to NaCN were retained after synaptic blockade with low Ca(2+)-high Mg(2+) or TTX. Thus hypoxic excitation 1) is maintained in cell culture, 2) is an intrinsic response, and 3) is likely due to the increase in a cation current. These hypoxia-excited neurons are likely candidates to function as central oxygen sensors. PMID- 10846052 TI - Suppression of acute viremia by short-term postexposure prophylaxis of simian/human immunodeficiency virus SHIV-RT-infected monkeys with a novel reverse transcriptase inhibitor (GW420867) allows for development of potent antiviral immune responses resulting in efficient containment of infection. AB - A nonnucleoside reverse transcriptase (RT) inhibitor, GW420867, was tested for postexposure prophylaxis (PEP) in rhesus macaques experimentally infected with 100 50% tissue culture infective doses of a chimeric simian/human immunodeficiency virus (SHIV) containing the RT gene of HIV-1 (SHIV-RT). Animals were either mock treated, or treated for 4 weeks starting at 8 or 24 h postinfection (p.i.) with GW420867. While such therapy led to undetectable plasma viremia in three of six monkeys, a transient plasma viremia was noted in the other three treated animals at 2 to 4 weeks following cessation of therapy. Following this transient viremia all drug-treated animals showed low or undetectable levels of plasma viremia up to the last sample examined at 90 weeks p.i. Despite low and/or undetectable viremia, virus-specific cytotoxic T lymphocyte and viral Env-specific proliferative responses were seen in the peripheral blood mononuclear cells of both mock- and drug-treated animals as early as 3 weeks p.i. Such virus-specific cellular responses, however, were better maintained in the drug-treated animals than the mock-treated animals. In contrast to the virus-specific cellular response, the magnitude and kinetics of virus specific humoral responses appeared to correlate with the detection of viremia. These data support the view that a short-term PEP with GW420867 permits the generation and maintenance of long-lasting virus-specific cell-mediated immune responses while markedly reducing viral loads to undetectable levels for a prolonged period of time (90 weeks) and leads to long-term disease protection. This model provides a unique means to define mechanisms and correlates of disease protection. PMID- 10846053 TI - Feline leukemia virus envelope sequences that affect T-cell tropism and syncytium formation are not part of known receptor-binding domains. AB - The envelope protein is a primary pathogenic determinant for T-cell-tropic feline leukemia virus (FeLV) variants, the best studied of which is the immunodeficiency inducing virus, 61C. We have previously demonstrated that T-cell-tropic, cytopathic, and syncytium-inducing viruses evolve in cats infected with a relatively avirulent, transmissible form of FeLV, 61E. The envelope gene of an 81T variant, which encoded scattered single-amino-acid changes throughout the envelope as well as a 4-amino-acid insertion in the C-terminal half of the surface unit (SU) of envelope, was sufficient to confer the T-cell-tropic, cytopathic phenotype (J. L. Rohn, M. S. Moser, S. R. Gwynn, D. N. Baldwin, and J. Overbaugh, J. Virol. 72:2686-2696, 1998). In the present study, we examined the role of the 4-amino-acid insertion in determining viral replication and tropism of FeLV-81T. The 4-amino-acid insertion was found to be functionally equivalent to a 6-amino-acid insertion at an identical location in the 61C variant. However, viruses expressing a chimeric 61E/81T SU, containing the insertion together with the N terminus of 61E SU, were found to be replication defective and were impaired in the processing of the envelope precursor into the functional SU and transmembrane (TM) proteins. In approximately 10% of cultured feline T cells (3201) transfected with the 61E/81T envelope chimeras and maintained over time, replication-competent tissue culture-adapted variants were isolated. Compensatory mutations in the SU of the tissue culture-adapted viruses were identified at positions 7 and 375, and each was shown to restore envelope protein processing when combined with the C-terminal 81T insertion. Unexpectedly, these viruses displayed different phenotypes in feline T cells: the virus with a change from glutamine to proline at position 7 acquired a T-cell-tropic, cytopathic phenotype, whereas the virus with a change from valine to leucine at position 375 had slower replication kinetics and caused no cytopathic effects. Given the differences in the replication properties of these viruses, it is noteworthy that the insertion as well as the two single-amino-acid changes all occur outside of predicted FeLV receptor-binding domains. PMID- 10846054 TI - Asynchronous accumulation of lettuce infectious yellows virus RNAs 1 and 2 and identification of an RNA 1 trans enhancer of RNA 2 accumulation. AB - Time course and mutational analyses were used to examine the accumulation in protoplasts of progeny RNAs of the bipartite Crinivirus, Lettuce infectious yellow virus (LIYV; family Closteroviridae). Hybridization analyses showed that simultaneous inoculation of LIYV RNAs 1 and 2 resulted in asynchronous accumulation of progeny LIYV RNAs. LIYV RNA 1 progeny genomic and subgenomic RNAs could be detected in protoplasts as early as 12 h postinoculation (p.i.) and accumulated to high levels by 24 h p.i. The LIYV RNA 1 open reading frame 2 (ORF 2) subgenomic RNA was the most abundant of all LIYV RNAs detected. In contrast, RNA 2 progeny were not readily detected until ca. 36 h p.i. Mutational analyses showed that in-frame stop codons introduced into five of seven RNA 2 ORFs did not affect accumulation of progeny LIYV RNA 1 or RNA 2, confirming that RNA 2 does not encode proteins necessary for LIYV RNA replication. Mutational analyses also supported that LIYV RNA 1 encodes proteins necessary for replication of LIYV RNAs 1 and 2. A mutation introduced into the LIYV RNA 1 region encoding the overlapping ORF 1B and ORF 2 was lethal. However, mutations introduced into only LIYV RNA 1 ORF 2 resulted in accumulation of progeny RNA 1 near or equal to wild type RNA 1. In contrast, the RNA 1 ORF 2 mutants did not efficiently support the trans accumulation of LIYV RNA 2. Three distinct RNA 1 ORF 2 mutants were analyzed and all exhibited a similar phenotype for progeny LIYV RNA accumulation. These data suggest that the LIYV RNA 1 ORF 2 encodes a trans enhancer for RNA 2 accumulation. PMID- 10846055 TI - In vivo induction of a high-avidity, high-frequency cytotoxic T-lymphocyte response is associated with antiviral protective immunity. AB - Many approaches are currently being developed to deliver exogenous antigen into the major histocompatibility complex class I-restricted antigen pathway, leading to in vivo priming of CD8(+) cytotoxic T cells. One attractive possibility consists of targeting the antigen to phagocytic or macropinocytic antigen presenting cells. In this study, we demonstrate that strong CD8(+) class I restricted cytotoxic responses are induced upon intraperitoneal immunization of mice with different peptides, characterized as CD8(+) T-cell epitopes, bound to 1 microm synthetic latex microspheres and injected in the absence of adjuvant. The cytotoxic response induced against a lymphocytic choriomeningitis virus (LCMV) peptide linked to these microspheres was compared to the cytotoxic T-lymphocyte (CTL) response obtained upon immunization with the nonreplicative porcine parvovirus-like particles (PPV:VLP) carrying the same peptide (PPV:VLP-LCMV) previously described (C. Sedlik, M. F. Saron, J. Sarraseca, I. Casal, and C. Leclerc, Proc. Natl. Acad. Sci. USA 94:7503-7508, 1997). We show that the induction of specific CTL activity by peptides bound to microspheres requires CD4(+) T-cell help in contrast to the CTL response obtained with the peptide delivered by viral pseudoparticles. Furthermore, PPV:VLP are 100-fold more efficient than microspheres in generating a strong CTL response characterized by a high frequency of specific T cells of high avidity. Moreover, PPV:VLP-LCMV are able to protect mice against a lethal LCMV challenge whereas microspheres carrying the LCMV epitope fail to confer such protection. This study demonstrates the crucial involvement of the frequency and avidity of CTLs in conferring antiviral protective immunity and highlights the importance of considering these parameters when developing new vaccine strategies. PMID- 10846056 TI - The long terminal repeat of Jaagsiekte sheep retrovirus is preferentially active in differentiated epithelial cells of the lungs. AB - Jaagsiekte sheep retrovirus (JSRV) is the etiologic agent of a contagious bronchioloalveolar carcinoma of sheep known as sheep pulmonary adenomatosis (SPA; ovine pulmonary carcinoma). JSRV is unique among retroviruses because it transforms the alveolar type II cells and the nonciliated bronchiolar cells (Clara cells) of the lungs; these cells are where JSRV is specifically expressed in both naturally and experimentally SPA-affected sheep. In this study, we investigated the cell specificity of JSRV expression. By transient-transfection assays of 23 different cell lines with a reporter plasmid driven by the JSRV long terminal repeat (LTR), pJS21-luc, we found that the JSRV LTR is preferentially active in cell lines derived from type II pneumocytes and Clara cells (MLE-15 and mtCC1-2 mouse cell lines). Reporter assays using progressive 5' deletions of pJS21-luc allowed us to establish that the JSRV enhancers are able to activate the JSRV proximal promoter in MLE-15 and mtCC1-2 cells, but they have very low activity in mouse cells of other lineages (e.g., NIH 3T3). The JSRV enhancers are able to activate heterologous promoters in both MLE-15 and 3T3 cells, although optimal activity is achieved in MLE-15 cells only with the homologous JSRV promoter. Thus, JSRV cell-specific LTR activity appears to result from an interaction between the enhancer elements and the JSRV proximal promoter elements. By mutation analysis, we established that an upstream NF-kappaB-like element appears to be responsible for approximately 50% of the JSRV LTR transcriptional activity in MLE-15 cells. Electrophoretic mobility shift assays showed evidence of a factor(s) that binds to this sequence. Antibody supershift experiments indicated that the factor(s) is not related to NF-kappaB component p50 or p52. This factor also appeared to be present in cells that do not support a high level of JSRV expression. Finally the JSRV(21) LTR contains putative enhancer binding motifs for transcription factors such as hepatocyte nuclear factor 3 (HNF-3) that are involved in lung-specific gene expression. Cotransfection experiments demonstrated that exogenous HNF-3 is able to enhance the expression of pJS21-luc in NIH 3T3 cells, which normally show minimal enhancer activity for the JSRV LTR. PMID- 10846057 TI - Association of RNA polymerase complexes of the parasitic protozoan Cryptosporidium parvum with virus-like particles: heterogeneous system. AB - RNA polymerase complexes were purified from Cryptosporidium parvum, a parasitic protozoan known to infect many species of mammals including humans. Western blot analysis revealed the association of the complexes with two different proteins, encoded by large and small segments of viral double-stranded RNAs. Each complex was found to contain only double-stranded RNA, both double- and single-stranded RNA, or only single-stranded RNA. Maximum RNA-dependent RNA polymerase activity was observed within the complexes containing both double- and single-stranded RNAs. These complexes possessed both transcriptase and replicase polymerase activities. Virus-like particles with a diameter of 31 nm were copurified with RNA polymerase complexes, and buoyant density and polymerase studies suggest that C. parvum harbors a putative double-stranded RNA virus which separately encapsidates the large and small RNA segments. The mechanism of replication and other characteristics of this virus are similar to those of the viruses of the family Partitiviridae, previously identified only in fungi and plants. PMID- 10846058 TI - Differential pathogenicity of two feline leukemia virus subgroup A molecular clones, pFRA and pF6A. AB - F6A, a molecular clone of subgroup A feline leukemia virus (FeLV) is considered to be highly infectious but weakly pathogenic. In recent studies with a closely related subgroup A molecular clone, FRA, we demonstrated high pathogenicity and a strong propensity to undergo recombination with endogenous FeLV (enFeLV), leading to a high frequency of transition from subgroup A to A/B. The present study was undertaken to identify mechanisms of FeLV pathogenesis that might become evident by comparing the two closely related molecular clones. F6A was shown to have an infectivity similar to that of FRA when delivered as a provirus. Virus load and antibody responses were also similar, although F6A-infected cats consistently carried higher virus loads than FRA-infected cats. However, F6A-infected cats were slower to undergo de novo recombination with enFeLV and showed slower progression to disease than FRA-infected cats. Tumors collected from nine pF6A- or pFRA-inoculated cats expressed lymphocyte markers for T cells (seven tumors) and B cells (one tumor), and non-T/B cells (one tumor). One cat with an A-to-A/C conversion developed erythrocyte hypoplasia. Genomic mapping of recombinants from pF6A- and pFRA-inoculated cats revealed similar crossover sites, suggesting that the genomic makeup of the recombinants did not contribute to increased progression to neoplastic disease. From these studies, the mechanism most likely to account for the pathologic differences between F6A and FRA is the lower propensity for F6A to undergo de novo recombination with enFeLV in vivo. A lower recombination rate is predicted to slow the transition from subgroup A to A/B and slow the progression to disease. PMID- 10846059 TI - Enhanced binding of antibodies to neutralization epitopes following thermal and chemical inactivation of human immunodeficiency virus type 1. AB - Inactivation of viral particles is the basis for several vaccines currently in use. Initial attempts to use simian immunodeficiency virus to model a killed human immunodeficiency virus type 1 (HIV-1) vaccine were unsuccessful, and limited subsequent effort has been directed toward a systematic study of the requirements for a protective killed HIV-1 vaccine. Recent insights into HIV-1 virion and glycoprotein structure and neutralization epitopes led us to revisit whether inactivated HIV-1 particles could serve as the basis for an HIV-1 vaccine. Our results indicate that relatively simple processes involving thermal and chemical inactivation can inactivate HIV-1 by at least 7 logs. For some HIV-1 strains, significant amounts of envelope glycoproteins are retained in high molecular-weight fractions. Importantly, we demonstrate retention of each of three conformation-dependent neutralization epitopes. Moreover, reactivity of monoclonal antibodies directed toward these epitopes is increased following treatment, suggesting greater exposure of the epitopes. In contrast, treatment of free envelope under the same conditions leads only to decreased antibody recognition. These inactivated virions can also be presented by human dendritic cells to direct a cell-mediated immune response in vitro. These data indicate that a systematic study of HIV-1 inactivation, gp120 retention, and epitope reactivity with conformation-specific neutralizing antibodies can provide important insights for the development of an effective killed HIV-1 vaccine. PMID- 10846061 TI - The E4-6/7 protein functionally compensates for the loss of E1A expression in adenovirus infection. AB - The E1A gene products are required and sufficient for activation of adenovirus gene expression in cultured cells. The E4-6/7 gene product induces the binding of the cellular transcription factor E2F to the viral E2a promoter region. The induction of E2F binding to the E2a promoter in vitro is directly correlated with transcriptional activation of the E2a promoter in vivo. The E2 region encodes the viral replication proteins, yet adenoviruses lacking E4-6/7 function demonstrate no defective phenotype in infected cells. Here we show that the E4-6/7 protein can functionally compensate for E1A expression in virus infection. In the absence of the E1A gene products, expression of the E4-6/7 protein is sufficient to displace retinoblastoma protein family members from E2Fs, activate expression of early region 2 via induction of E2F DNA binding to the E2a promoter region, and significantly enhance replication of an E1A-defective adenovirus. These results have implications in the regulation of viral gene expression and for the development of recombinant adenovirus vectors. PMID- 10846060 TI - Transforming growth factor beta 1 stimulates expression of the Epstein-Barr virus BZLF1 immediate-early gene product ZEBRA by an indirect mechanism which requires the MAPK kinase pathway. AB - Disruption of Epstein-Barr virus (EBV) latency is mediated by ZEBRA, the protein product of the immediate-early EBV gene, BZLF1. In vitro, phorbol 12-myristate 13 acetate (PMA), a potent activator of protein kinase C (PKC), induces reactivation of EBV. However, the physiological stimuli responsible for the disruption of viral latency are not well characterized. Transforming growth factor beta 1 (TGF beta1) has also been shown to trigger the reactivation of EBV in Burkitt lymphoma cell lines; however, the effect of TGF-beta1 on ZEBRA expression has not been reported. To further understand this phenomenon, we have investigated the effect of TGF-beta1 on ZEBRA expression. Our results indicate that the treatment of different EBV-positive Burkitt's lymphoma cell lines with TGF-beta1 induces a time-dependent activation of BZLF1 transcription with a corresponding increase in the production of the protein ZEBRA. TGF-beta1 has been shown to exert its effects through a wide range of intracellular routes; in the present study, we have explored these pathways. Transient expression of Smad proteins on their own had no effect on ZEBRA expression. A specific inhibitor of p38 mitogen-activated protein kinase (MAPK), SB203580, did not affect TGF-beta1-induced ZEBRA expression, whereas treatment with the MAPK/ERK kinase inhibitors, PD98059 and U0126, dramatically decreased this induction. This suggests that TGF-beta1 effect on BZLF1 expression requires the MAPK pathway. However, in Raji and B95-8 cells additional routes can be used, as (i) the inhibition of ZEBRA induction by PD98059 or U0126 was incomplete, whereas these inhibitors completely abolished PMA-induced ZEBRA expression, (ii) TGF-beta1 induction of ZEBRA expression occurs in PKC-depleted cells, (iii) in Raji and in B95-8 cells, the effect of TGF-beta1 and PMA are additive. Transient transfection of the EBV-negative B-cell line DG75 with a BZLF1 promoter-fusion construct (Zp-CAT) showed that under conditions where the BZLF1 promoter is activated by PMA treatment, TGF-beta1 had no significant effect on the expression of the chloramphenicol acetyltransferase gene. Furthermore, TGF-beta1 induction of BZLF1 transcripts is dependent on de novo protein synthesis, which suggests that TGF-beta1 induces BZLF1 expression by an indirect mechanism. PMID- 10846062 TI - A stem-loop motif formed by the immediate 5' terminus of the bovine viral diarrhea virus genome modulates translation as well as replication of the viral RNA. AB - Bovine viral diarrhea virus (BVDV), a Pestivirus member of the Flaviviridae family, has a positive-stranded RNA genome which consists of a single open reading frame (ORF) and untranslated regions (UTRs) at the 5' and 3' ends. The 5' UTR harbors extensive RNA structure motifs; most of them were shown to contribute to an internal ribosomal entry site (IRES), which mediates cap-independent translation of the ORF. The extreme 5'-terminal region of the BVDV genome had so far been believed not to be required for IRES function. By structure probing techniques, we initially verified the existence of a computer-predicted stem-loop motif at the 5' end of the viral genome (hairpin Ia) as well as at the 3' end of the complementary negative-strand replication intermediate [termed hairpin Ia ( )]. While the stem of this structure is mainly constituted of nucleotides that are conserved among pestiviruses, the loop region is predominantly composed of variable residues. Taking a reverse genetics approach to a subgenomic BVDV replicon RNA (DI9c) which could be equally employed in a translation as well as replication assay system based on BHK-21 cells, we obtained the following results. (i) Proper folding of the Ia stem was found to be crucial for efficient translation. Thus, in the context of an authentic replication-competent viral RNA, the 5'-terminal motif operates apparently as an integral functional part of the ribosome entry. (ii) An intact loop structure and a stretch of nucleotide residues that constitute a portion of the stem of the Ia or the Ia (-) motif, respectively, were defined to represent important determinants of the RNA replication pathway. (iii) Formation of the stem structure of the Ia (-) motif was determined to be not critical for RNA replication. In summary, our findings affirmed that the 5'-terminal region of the BVDV genome encodes a bifunctional secondary structure motif which may enable the viral RNA to switch from the translation to the replicative cycle and vice versa. PMID- 10846063 TI - The intracytoplasmic domain of the Env transmembrane protein is a locus for attenuation of simian immunodeficiency virus SIVmac in rhesus macaques. AB - The human and simian immunodeficiency virus (HIV-1 and SIVmac) transmembrane proteins contain unusually long intracytoplasmic domains (ICD-TM). These domains are suggested to play a role in envelope fusogenicity, interaction with the viral matrix protein during assembly, viral infectivity, binding of intracellular calmodulin, disruption of membranes, and induction of apoptosis. Here we describe a novel mutant virus, SIVmac-M4, containing multiple mutations in the coding region for the ICD-TM of pathogenic molecular clone SIVmac239. Parental SIVmac239 Nef+ produces high-level persistent viremia and simian AIDS in both juvenile and newborn rhesus macaques. The ICD-TM region of SIVmac-M4 contains three stop codons, a +1 frameshift, and mutation of three highly conserved, charged residues in the conserved C-terminal alpha-helix referred to as lentivirus lytic peptide 1 (LLP-1). Overlapping reading frames for tat, rev, and nef are not affected by these changes. In this study, four juvenile macaques received SIVmac-M4 by intravenous injection. Plasma viremia, as measured by branched-DNA (bDNA) assay, reached a peak at 2 weeks postinoculation but dropped to below detectable levels by 12 weeks. At over 1.5 years postinoculation, all four juvenile macaques remain healthy and asymptomatic. In a subsequent experiment, four neonatal rhesus macaques were given SIVmac-M4 intravenously. These animals exhibited high levels of viremia in the acute phase (2 weeks postinoculation) but are showing a relatively low viral load in the chronic phase of infection, with no clinical signs of disease for 1 year. These findings demonstrated that the intracytoplasmic domain of the transmembrane Env (Env-TM) is a locus for attenuation in rhesus macaques. PMID- 10846064 TI - Kinetic analysis of human immunodeficiency virus type 1 assembly reveals the presence of sequential intermediates. AB - The assembly and budding of lentiviruses, such as human immunodeficiency virus type 1 (HIV-1), are mediated by the Gag protein precursor, but the molecular details of these processes remain poorly defined. In this study, we have combined pulse-chase techniques with density gradient centrifugation to identify, isolate, and characterize sequential kinetic intermediates in the lentivirus assembly process. We show that newly synthesized HIV-1 Gag rapidly forms cytoplasmic protein complexes that are resistant to detergent treatment, sensitive to protease digestion, and degraded intracellularly. A subpopulation of newly synthesized Gag binds membranes within 5 to 10 min and over several hours assembles into membrane-bound complexes of increasing size and/or density that can be resolved on Optiprep density gradients. These complexes likely represent assembly intermediates because they are not observed with assembly-defective Gag mutants and can be chased into extracellular viruslike particles. At steady state, nearly all of the Gag is present as membrane-bound complexes in various stages of assembly. The identification of sequential assembly intermediates provides the first demonstration that HIV-1 particle assembly proceeds via an ordered process. Assembly intermediates should serve as attractive targets for the design of antiviral agents that interfere with the process of particle production. PMID- 10846065 TI - Human parechovirus 1 utilizes integrins alphavbeta3 and alphavbeta1 as receptors. AB - Human parechovirus 1 (HPEV1) displays an arginine-glycine-aspartic acid (RGD) motif in the VP1 capsid protein, suggesting integrins as candidate receptors for HPEV1. A panel of monoclonal antibodies (MAbs) specific for integrins alphavbeta3, alphavbeta1, and alphavbeta5, which have the ability to recognize the RGD motif, and also a MAb specific for integrin alpha2beta1, an integrin that does not recognize the RGD motif, were tested on A549 cells. Our results showed that integrin alphav-specific MAb reduced infectivity by 85%. To specify which alphav integrins the virus utilizes, we tested MAbs specific to integrins alphavbeta3 and alphavbeta1 which reduced infectivity significantly, while a MAb specific for integrin alphavbeta5, as well as the MAb specific for alpha2beta1, showed no reduction. When a combination of MAbs specific for integrins alphavbeta3 and alphavbeta1 were used, virus infectivity was almost completely inhibited; this shows that integrins alphavbeta3 and alphavbeta1 are utilized by the virus. We therefore proceeded to test whether alphav integrins' natural ligands fibronectin and vitronectin had an effect on HPEV1 infectivity. We found that vitronectin reduced significantly HPEV1 infectivity, whereas a combination of vitronectin and fibronectin abolished infection. To verify the use of integrins alphavbeta3 and alphavbeta1 as HPEV1 receptors, CHO cells transfected and expressing either integrin alphavbeta3 or integrin alphavbeta1 were used. It was shown that the virus could successfully infect these cells. However, in immunoprecipitation experiments using HPEV1 virions and allowing the virus to bind to solubilized A549 cell extract, we isolated and confirmed by Western blotting the alphavbeta3 heterodimer. In conclusion, we found that HPEV1 utilises both integrin alphavbeta3 and alphavbeta1 as receptors; however, in cells that express both integrins, HPEV1 may preferentially bind integrin alphavbeta3. PMID- 10846066 TI - Analysis of cellular factors that mediate nuclear export of RNAs bearing the Mason-Pfizer monkey virus constitutive transport element. AB - There is now convincing evidence that the human Tap protein plays a critical role in mediating the nuclear export of mRNAs that contain the Mason-Pfizer monkey virus constitutive transport element (CTE) and significant evidence that Tap also participates in global poly(A)(+) RNA export. Previously, we had mapped carboxy terminal sequences in Tap that serve as an essential nucleocytoplasmic shuttling domain, while others had defined an overlapping Tap sequence that can bind to the FG repeat domains of certain nucleoporins. Here, we demonstrate that these two biological activities are functionally correlated. Specifically, mutations in Tap that block nucleoporin binding also block both nucleocytoplasmic shuttling and the Tap-dependent nuclear export of CTE-containing RNAs. In contrast, mutations that do not inhibit nucleoporin binding also fail to affect Tap shuttling. Together, these data indicate that Tap belongs to a novel class of RNA export factors that can target bound RNA molecules directly to the nuclear pore without the assistance of an importin beta-like cofactor. In addition to nucleoporins, Tap has also been proposed to interact with a cellular cofactor termed p15. Although we were able to confirm that Tap can indeed bind p15 specifically both in vivo and in vitro, a mutation in Tap that blocked p15 binding only modestly inhibited CTE-dependent nuclear RNA export. However, p15 did significantly enhance the affinity of Tap for the CTE in vitro and readily formed a ternary complex with Tap on the CTE. This result suggests that p15 may play a significant role in the recruitment of the Tap nuclear export factor to target RNA molecules in vivo. PMID- 10846067 TI - AMF-1/Gps2 binds p300 and enhances its interaction with papillomavirus E2 proteins. AB - The cellular protein AMF-1 (Gps2) positively modulates gene expression by the papillomavirus E2 protein (D. E. Breiding et al., Mol. Cell. Biol. 17:7208-7219, 1997). We show here that AMF-1 also binds the transcriptional coactivator p300 in vitro and in vivo. E2 interacted weakly with p300. These observations led to a model in which AMF-1 recruits p300 into a complex with E2. Cotransfection of AMF 1 or p300 stimulated levels of E2-dependent transcription, while cotransfection of both AMF-1 and p300 showed an additive effect. The functional significance of p300 recruitment for E2 transactivation was evidenced by repression of E2 activated transcription by adenovirus E1A, which inhibits both coactivator and acetylase activities of p300. Antibodies to AMF-1 or E2 immunoprecipitated histone acetylase activity from cell lysates. Western blotting using antibody against acetyl-lysine failed to detect acetylation of AMF-1 or E2 in complex with p300. These results suggest that AMF-1 facilitates the recruitment of p300 and its histone acetylase activity into complexes with E2 and represents a novel mechanism of transcriptional activation. PMID- 10846068 TI - The Cys(3)-His(1) motif of the respiratory syncytial virus M2-1 protein is essential for protein function. AB - The M2 gene of respiratory syncytial (RS) virus has two open reading frames (ORFs). ORF1 encodes a 22-kDa protein termed M2-1. The M2-1 protein contains a Cys(3)-His(1) motif (C-X(7)-C-X(5)-C-X(3)-H) near the amino terminus. This motif is conserved in all human, bovine, and ovine strains of RS virus. A similar motif found in the mammalian transcription factor Nup475 has been shown to bind zinc. The M2-1 protein of human RS virus functions as a transcription factor which increases polymerase processivity, and it enhances readthrough of intergenic junctions during RS virus transcription, thereby acting as a transcription antiterminator. The M2-1 protein also interacts with the nucleocapsid protein. We examined the effects of mutations of cysteine and histidine residues predicted to coordinate zinc in the Cys(3)-His(1) motif on transcription antitermination and N protein binding. We found that mutating the predicted zinc-coordinating residues, the cysteine residues at amino acid positions 7 and 15 and the histidine residue at position 25, prevented M2-1 from enhancing transcriptional readthrough. In contrast, mutations of amino acids within this motif not predicted to coordinate zinc had no effect. Mutations of the predicted zinc-coordinating residues in the Cys(3)-His(1) motif also prevented M2-1 from interacting with the nucleocapsid protein. One mutation of a noncoordinating residue in the motif which did not affect readthrough during transcription, E10G, prevented interaction with the nucleocapsid protein. This suggests that M2-1 does not require interaction with the nucleocapsid protein in order to function during transcription. Analysis of the M2-1 protein in reducing sodium dodecyl sulfate-polyacrylamide gels revealed two major forms distinguished by their mobilities. The slower migrating form was shown to be phosphorylated, whereas the faster migrating form was not. Mutations in the Cys(3)-His(1) motif caused a change in distribution of the M2-1 protein from the slower to the faster migrating form. The data presented here show that the Cys(3)-His(1) motif of M2-1 is essential for maintaining the functional integrity of the protein. PMID- 10846069 TI - Role of NH(2)- and COOH-terminal domains of the P protein of human parainfluenza virus type 3 in transcription and replication. AB - The phosphoproteins (P proteins) of paramyxoviruses play a central role in transcription and replication of the viruses by forming the RNA polymerase complex L-P and encapsidation complex (N-P) with nucleocapsid protein (N) and binding to N protein-encapsidated genome RNA template (N-RNA template). We have analyzed the human parainfluenza virus type 3 (HPIV3) P protein and deletion mutants thereof in an in vitro transcription and in vivo replication system. The in vitro system utilizes purified N-RNA template and cell extract containing L and P proteins coexpressed via plasmids using a recombinant vaccinia virus expression system. The in vivo system takes advantage of minigenome replication, which measures luciferase reporter gene expression from HPIV3 minigenomes by viral proteins in a recombinant vaccinia virus expression system. These studies revealed that the C-terminal 20-amino-acid region of P is absolutely required for transcription in vitro and luciferase expression in vivo, suggesting its critical role in viral RNA synthesis. The N-terminal 40-amino-acid region, on the other hand, is essential for luciferase expression but dispensable for transcription in vitro. Consistent with these findings, the C-terminal domain is required for binding of P protein to the N-RNA template involved in both transcription and replication, whereas the N-terminal domain is required for the formation of soluble N-P complex involved in encapsidation of nascent RNA chains during replication. Coimmunoprecipitation analysis showed that the P protein forms a stable homooligomer (perhaps a trimer) that is present in L-P and N-P complexes in the higher oligomeric forms (at least a pentamer). Interestingly, coexpression of a large excess of N- or C-terminally deleted P with wild-type P had no effect on minigenome replication in vivo, notwithstanding the formation of heterooligomeric complexes. These data indicate that P protein with a deleted terminal domain can function normally within the P heterooligomeric complex to carry out transcription and replication in vivo. PMID- 10846071 TI - Optimization of a weak 3' splice site counteracts the function of a bovine papillomavirus type 1 exonic splicing suppressor in vitro and in vivo. AB - Alternative splicing is a critical component of the early to late switch in papillomavirus gene expression. In bovine papillomavirus type 1 (BPV-1), a switch in 3' splice site utilization from an early 3' splice site at nucleotide (nt) 3225 to a late-specific 3' splice site at nt 3605 is essential for expression of the major capsid (L1) mRNA. Three viral splicing elements have recently been identified between the two alternative 3' splice sites and have been shown to play an important role in this regulation. A bipartite element lies approximately 30 nt downstream of the nt 3225 3' splice site and consists of an exonic splicing enhancer (ESE), SE1, followed immediately by a pyrimidine-rich exonic splicing suppressor (ESS). A second ESE (SE2) is located approximately 125 nt downstream of the ESS. We have previously demonstrated that the ESS inhibits use of the suboptimal nt 3225 3' splice site in vitro through binding of cellular splicing factors. However, these in vitro studies did not address the role of the ESS in the regulation of alternative splicing. In the present study, we have analyzed the role of the ESS in the alternative splicing of a BPV-1 late pre-mRNA in vivo. Mutation or deletion of just the ESS did not significantly change the normal splicing pattern where the nt 3225 3' splice site is already used predominantly. However, a pre-mRNA containing mutations in SE2 is spliced predominantly using the nt 3605 3' splice site. In this context, mutation of the ESS restored preferential use of the nt 3225 3' splice site, indicating that the ESS also functions as a splicing suppressor in vivo. Moreover, optimization of the suboptimal nt 3225 3' splice site counteracted the in vivo function of the ESS and led to preferential selection of the nt 3225 3' splice site even in pre-mRNAs with SE2 mutations. In vitro splicing assays also showed that the ESS is unable to suppress splicing of a pre-mRNA with an optimized nt 3225 3' splice site. These data confirm that the function of the ESS requires a suboptimal upstream 3' splice site. A surprising finding of our study is the observation that SE1 can stimulate both the first and the second steps of splicing. PMID- 10846070 TI - Additive effect of neutralizing antibody and antiviral drug treatment in preventing virus escape and persistence. AB - Poorly cytopathic or noncytopathic viruses can escape immune surveillance and establish a chronic infection. Here we exploited the strategy of combining antiviral drug treatment with the induction of a neutralizing antibody response to avoid the appearance of neutralization-resistant virus variants. Despite the fact that H25 immunoglobulin transgenic mice infected with lymphocytic choriomeningitis virus mounted an early neutralizing antibody response, the virus escaped from neutralization and persisted. After ribavirin treatment of H25 transgenic mice, the appearance of neutralization-resistant virus was prevented and virus was cleared. Thus, the combination of virus-neutralizing antibodies and chemotherapy efficiently controlled the infection, whereas each defense line alone did not. Similar additive effects may be unexpectedly efficient and beneficial in humans after infections with persistent viruses such as hepatitis C virus and hepatitis B virus and possibly human immunodeficiency virus. PMID- 10846072 TI - The core of the respiratory syncytial virus fusion protein is a trimeric coiled coil. AB - Entry into the host cell by enveloped viruses is mediated by fusion (F) or transmembrane glycoproteins. Many of these proteins share a fold comprising a trimer of antiparallel coiled-coil heterodimers, where the heterodimers are formed by two discontinuous heptad repeat motifs within the proteolytically processed chain. The F protein of human respiratory syncytial virus (RSV; the major cause of lower respiratory tract infections in infants) contains two corresponding regions that are predicted to form coiled coils (HR1 and HR2), together with a third predicted heptad repeat (HR3) located in a nonhomologous position. In order to probe the structures of these three domains and ascertain the nature of the interactions between them, we have studied the isolated HR1, HR2, and HR3 domains of RSV F by using a range of biophysical techniques, including circular dichroism, nuclear magnetic resonance spectroscopy, and sedimentation equilibrium. HR1 forms a symmetrical, trimeric coiled coil in solution (K(3) approximately 2.2 x 10(11) M(-2)) which interacts with HR2 to form a 3:3 hexamer. The HR1-HR2 interaction domains have been mapped using limited proteolysis, reversed-phase high-performance liquid chromatography, and electrospray-mass spectrometry. HR2 in isolation exists as a largely unstructured monomer, although it exhibits a tendency to form aggregates with beta-sheet-like characteristics. Only a small increase in alpha-helical content was observed upon the formation of the hexamer. This suggests that the RSV F glycoprotein contains a domain that closely resembles the core structure of the simian parainfluenza virus 5 fusion protein (K. A. Baker, R. E. Dutch, R. A. Lamb, and T. S. Jardetzky, Mol. Cell 3:309-319, 1999). Finally, HR3 forms weak alpha-helical homodimers that do not appear to interact with HR1, HR2, or the HR1-HR2 complex. The results of these studies support the idea that viral fusion proteins have a common core architecture. PMID- 10846073 TI - Structural, functional, and genetic comparisons of Epstein-Barr virus nuclear antigen 3A, 3B, and 3C homologues encoded by the rhesus lymphocryptovirus. AB - EBNA-3A, -3B, and -3C are three latent infection nuclear proteins important for Epstein-Barr virus (EBV)-induced B-cell immortalization and the immune response to EBV infection. All three are hypothesized to function as transcriptional transactivators, but little is known about their precise mechanism of action or their role in EBV pathogenesis. We have cloned and studied the three EBNA-3 homologues from a closely related lymphocryptovirus (LCV) which naturally infects rhesus monkeys. The rhesus LCV EBNA-3A, -3B, and -3C homologues have 37, 40, and 36% amino acid identity with the EBV genes, respectively. Function, as measured by in vitro assays, also appears to be conserved with the EBV genes, since the rhesus LCV EBNA-3s can interact with the transcription factor RBP-Jkappa and the rhesus LCV EBNA-3C encodes a Q/P-rich domain with transcriptional activation properties. In order to better understand the relationship between these EBV and rhesus LCV latent infection genes, we asked if the rhesus LCV EBNA-3 locus could be recombined into the EBV genome and if it could substitute for the EBV EBNA-3s when assayed for human B-cell immortalization. Recombination between the EBV genome and rhesus LCV DNA was reasonably efficient. However, these studies suggest that the rhesus LCV EBNA-3 locus was not completely interchangeable with the EBV EBNA-3 locus for B-cell immortalization and that at least one determinant of the species restriction for LCV-induced B-cell immortalization maps to the EBNA-3 locus. The overall conservation of EBNA-3 structure and function between EBV and rhesus LCV indicates that rhesus LCV infection of rhesus monkeys can provide an important animal model for studying the role of the EBNA-3 genes in LCV pathogenesis. PMID- 10846074 TI - Binding of hepatitis C virus E2 glycoprotein to CD81 does not correlate with species permissiveness to infection. AB - Hepatitis C virus (HCV) glycoprotein E2 binds to human cells by interacting with the CD81 molecule, which has been proposed to be the viral receptor. A correlation between binding to CD81 and species permissiveness to HCV infection has also been reported. We have determined the sequence of CD81 from the tamarin, a primate species known to be refractory to HCV infection. Tamarin CD81 (t-CD81) differs from the human molecule at 5 amino acid positions (155, 163, 169, 180, and 196) within the large extracellular loop (LEL), where the binding site for E2 has been located. Soluble recombinant forms of human CD81 (h-CD81), t-CD81, and African green monkey CD81 (agm-CD81) LEL molecules were analyzed by enzyme-linked immunosorbent assay for binding to E2 glycoprotein. Both h-CD81 and t-CD81 molecules were able to bind E2. Competition experiments showed that the two receptors cross-compete and that the t-CD81 binds with stronger affinity than the human molecule. Recently, h-CD81 residue 186 has been characterized as the critical residue involved in the interaction with E2. Recombinant CD81 mutant proteins were expressed to test whether human and tamarin receptors interacted with E2 in a comparable manner. Mutation of residue 186 (F186L) dramatically reduced the binding capability of t-CD81, a result that has already been demonstrated for the human receptor, whereas the opposite mutation (L186F) in agm CD81 resulted in a neat gain of binding activity. Finally, the in vitro data were confirmed by detection of E2 binding to cotton-top tamarin (Saguinus oedipus) cell line B95-8 expressing endogenous CD81. These results indicate that the binding of E2 to CD81 is not predictive of an infection-producing interaction between HCV and host cells. PMID- 10846075 TI - Reovirus protein sigmaNS binds in multiple copies to single-stranded RNA and shares properties with single-stranded DNA binding proteins. AB - Reovirus nonstructural protein sigmaNS interacts with reovirus plus-strand RNAs in infected cells, but little is known about the nature of those interactions or their roles in viral replication. In this study, a recombinant form of sigmaNS was analyzed for in vitro binding to nucleic acids using gel mobility shift assays. Multiple units of sigmaNS bound to single-stranded RNA molecules with positive cooperativity and with each unit covering about 25 nucleotides at saturation. The sigmaNS protein did not bind preferentially to reovirus RNA over nonreovirus RNA in competition experiments but did bind preferentially to single stranded over double-stranded nucleic acids and with a slight preference for RNA over DNA. In addition, sigmaNS bound to single-stranded RNA to which a 19-base DNA oligonucleotide was hybridized at either end or near the middle. When present in saturative amounts, sigmaNS displaced this oligonucleotide from the partial duplex. The strand displacement activity did not require ATP hydrolysis and was inhibited by MgCl(2), distinguishing it from a classical ATP-dependent helicase. These properties of sigmaNS are similar to those of single-stranded DNA binding proteins that are known to participate in genomic DNA replication, suggesting a related role for sigmaNS in replication of the reovirus RNA genome. PMID- 10846076 TI - Characterization of the zinc binding activity of the rubella virus nonstructural protease. AB - The rubella virus (RUB) nonstructural (NS) protein (NSP) ORF encodes a protease that cleaves the NSP precursor (240 kDa) at a single site to produce two products. A cleavage site mutation was introduced into a RUB infectious cDNA clone and found to be lethal, demonstrating that cleavage of the NSP precursor is necessary for RUB replication. Based on computer alignments, the RUB NS protease was predicted to be a papain-like cysteine protease (PCP) with the residues Cys1152 and His1273 as the catalytic dyad; however, the RUB NS protease was recently found to require divalent cations such as Zn, Co, and Cd for activity (X. Liu, S. L. Ropp, R. J. Jackson, and T. K. Frey, J. Virol. 72:4463-4466, 1998). To analyze the function of metal cation binding in protease activity, Zn binding studies were performed using the minimal NS protease domain within the NSP ORF. When expressed as a maltose binding protein (MBP) fusion protein by bacteria, the NS protease exhibited activity both in the bacteria and in vitro following purification when denatured and refolded in the presence of Zn. Atomic absorption analysis detected 1.6 mol of Zn bound per mol of protein refolded in this manner. Expression of individual domains within the protease as MBP fusions and analysis by a Zn(65) binding assay revealed two Zn binding domains: one located at a predicted metal binding motif beginning at Cys1175 and the other one close to the cleavage site. Mutagenesis studies showed that Cys1175 and Cys1178 in the first domain and Cys1227 and His1273, the His in the predicted catalytic site, in the second domain are essential for zinc binding. All of these residues are also necessary for the protease activity, as were several other Cys residues not involved in Zn binding. Far-UV circular dichroism (CD) analysis of the MBP-NS protease fusion protein showed that the protease domain contained a large amount of alpha-helical structure, which is consistent with the results of secondary structural prediction. Both far-UV-CD and fluorescence studies suggested that Zn did not exert a major effect on the overall structure of the fusion protein. Finally, protease inhibitor assays found that the protease activity can be blocked by both metal ion chelators and the metalloprotease inhibitor captopril. In conjunction with the finding that the previously predicted catalytic site, His1273, is essential for zinc binding, this suggests that the RUB NS protease is actually a novel virus metalloprotease rather than a PCP. PMID- 10846077 TI - Optimized viral dose and transient immunosuppression enable herpes simplex virus ICP0-null mutants To establish wild-type levels of latency in vivo. AB - The reduced efficiency with which herpes simplex virus type 1 (HSV-1) mutants establish latent infections in vivo has been a fundamental obstacle in efforts to determine the roles of individual viral genes in HSV-1 reactivation. For example, in the absence of the "nonessential" viral immediate-early protein, ICP0, HSV-1 is severely impaired in its ability to (i) replicate at the site of inoculation and (ii) establish latency in neurons of the peripheral nervous system. The mouse ocular model of HSV latency was used in the present study to determine if the conditions of infection can be manipulated such that replication-impaired, ICP0 null mutants establish wild-type levels of latency, as measured by viral genome loads in latently infected trigeminal ganglia (TG). To this end, the effects of inoculum size and transient immunosuppression on the levels of acute replication in mouse eyes and of viral DNA in latently infected TG were examined. Following inoculation of mice with 2 x 10(3), 2 x 10(4), 2 x 10(5), or 2 x 10(6) PFU/eye, wild-type virus replicated in mouse eyes and established latency in TG with similar efficiencies at all four doses. In contrast, increasing the inoculum size of the ICP0-null mutants n212 and 7134 from 2 x 10(5) to 2 x 10(6) PFU/eye significantly decreased the levels of infectious virus detected in the tear films of mice from days 4 to 9 postinfection. In an attempt to establish the biological basis for this finding, the effect of viral dose on the induction of the host proinflammatory response was examined. Quantitative reverse transcription-PCR demonstrated that increasing the inoculum of 7134 from 2 x 10(4) to 2 x 10(6) PFU/eye significantly increased the expression of proinflammatory (interleukin 6), cell adhesion (intercellular adhesion molecule 1), and phagocyte-associated (CD11b) genes in mouse eyes 24 h postinfection. Furthermore, transient immunosuppression of mice with cyclophosphamide, but not cyclosporin A, significantly enhanced both the levels of acute n212 and 7134 replication in the eye and the levels of mutant viral genomes present in latently infected TG in a dose-dependent manner. Thus, the results of this study demonstrate that acute replication in the eye and the number of ICP0-null mutant genomes in latently infected TG can be increased to wild-type levels for both n212 and 7134 by (i) optimization of inoculum size and (ii) transient immunosuppression with cyclophosphamide. PMID- 10846078 TI - Immune responses and viral replication in long-term inapparent carrier ponies inoculated with equine infectious anemia virus. AB - Persistent infection of equids by equine infectious anemia virus (EIAV) is typically characterized by a progression during the first year postinfection from chronic disease with recurring disease cycles to a long-term asymptomatic infection that is maintained indefinitely. The goal of the current study was to perform a comprehensive longitudinal analysis of the course of virus infection and development of host immunity in experimentally infected horses as they progressed from chronic disease to long-term inapparent carriage. We previously described the evolution of EIAV genomic quasispecies (C. Leroux, C. J. Issel, and R. C. Montelaro, J. Virol. 71:9627-9639, 1997) and host immune responses (S. A. Hammond, S. J. Cook, D. L. Lichtenstein, C. J. Issel, and R. C. Montelaro, J. Virol. 71:3840-3852, 1997) in four experimentally infected ponies during sequential disease episodes associated with chronic disease during the first 10 months postinfection. In the current study, we extended the studies of these experimentally infected ponies to 3 years postinfection to characterize the levels of virus replication and development of host immune responses associated with the progression from chronic disease to long-term inapparent infection. The results of these studies revealed over a 10(3)-fold difference in the steady state levels of plasma viral RNA detected during long-term inapparent infection that correlated with the severity of chronic disease, indicating different levels of control of virus replication during long-term inapparent infections. Detailed analyses of antibody and cellular immune responses in all four ponies over the 3 year course of infection revealed a similar evolution during the first year postinfection of robust humoral and cellular immunity that then remained relatively constant during long-term inapparent infection. These observations indicate that immune parameters that have previously been correlated with EIAV vaccine protection fail to provide reliable immune correlates of control of virus replication or clinical outcome in experimental infections. Thus, these data emphasize the differences between immunity to virus exposure and immune control of an established viral infection and further emphasize the need to develop and evaluate novel immunoassays to define reliable immune correlates to vaccine and infection immunity, respectively. PMID- 10846079 TI - Cellular and viral specificities of human immunodeficiency virus type 1 vif protein. AB - The vif gene of human immunodeficiency virus type 1 (HIV-1) greatly enhances the infectivity of HIV-1 virions that are released from cells classified as nonpermissive (e.g., lymphocytes, macrophages, and H9 leukemic T cells) but is irrelevant in permissive cells (e.g., HeLa or COS cells). Recently, it was reported that vif expression in nonpermissive cells dramatically increases infectivity not only of HIV-1 but also of other enveloped viruses, including murine leukemia viruses (MLVs). This was surprising in part because MLVs and other murine retroviruses lack vif genes yet replicate efficiently in T lymphocytes. To investigate these issues, we first developed improved methods for producing substantial quantities of HIV-1 virions with vif deletions from healthy H9 cells. These virions had approximately the same amounts of major core proteins and envelope glycoproteins as the control wild-type virions but were only approximately 1% as infectious. We then produced H9 cells that contained wild type or vif deletion HIV-gpt proviruses, which lack a functional env gene. After superinfection with either xenotropic or amphotropic MLVs, these cells released HIV-gpt virions pseudotyped with an MLV envelope plus replication-competent MLV. Interestingly, the pseudotyped HIV-gpt (vif deletion) virions were noninfectious, whereas the MLV virions simultaneously released from the same H9 cells were fully infectious. These results strongly suggest that the Vif protein functions in a manner that is both cell specific and at least substantially specific for HIV-1 and related lentiviruses. In addition, these results confirm that vif deletion HIV-1 virions from nonpermissive cells are blocked at a postpenetration stage of the infection pathway. PMID- 10846080 TI - A positive-strand RNA virus with three very different subgenomic RNA promoters. AB - Numerous RNA viruses generate subgenomic mRNAs (sgRNAs) for expression of their 3'-proximal genes. A major step in control of viral gene expression is the regulation of sgRNA synthesis by specific promoter elements. We used barley yellow dwarf virus (BYDV) as a model system to study transcriptional control in a virus with multiple sgRNAs. BYDV generates three sgRNAs during infection. The sgRNA1 promoter has been mapped previously to a 98-nucleotide (nt) region which forms two stem-loop structures. It was determined that sgRNA1 is not required for BYDV RNA replication in oat protoplasts. In this study, we show that neither sgRNA2 nor sgRNA3 is required for BYDV RNA replication. The promoters for sgRNA2 and sgRNA3 synthesis were mapped by using deletion mutagenesis. The minimal sgRNA2 promoter is approximately 143 nt long (nt 4810 to 4952) and is located immediately downstream of the putative sgRNA2 start site (nt 4809). The minimal sgRNA3 core promoter is 44 nt long (nt 5345 to 5388), with most of the sequence located downstream of sgRNA3 start site (nt 5348). For both promoters, additional sequences upstream of the start site enhanced sgRNA promoter activity. These promoters contrast to the sgRNA1 promoter, in which almost all of the promoter is located upstream of the transcription initiation site. Comparison of RNA sequences and computer-predicted secondary structures revealed little or no homology between the three sgRNA promoter elements. Thus, a small RNA virus with multiple sgRNAs can have very different subgenomic promoters, which implies a complex system for promoter recognition and regulation of subgenomic RNA synthesis. PMID- 10846082 TI - Functional analysis of the genomic and antigenomic promoters of human respiratory syncytial virus. AB - The promoters involved in transcription and RNA replication by respiratory syncytial virus (RSV) were examined by using a plasmid-based minireplicon system. The 3' ends of the genome and antigenome, which, respectively, contain the 44 nucleotide (nt) leader (Le) and 155-nt trailer-complement (TrC) regions, should each contain a promoter for RNA replication. The 3' genome end also should have the promoter for transcription. Substitution for the Le with various lengths of TrC demonstrated that the 3'-terminal 36 nt of TrC are sufficient for extensive (but not maximal) replication and that when juxtaposed with a transcription gene start (GS) signal, this sequence was also able to direct transcription. It was also shown that the region of Le immediately preceding the GS signal of the first gene could be deleted with either no effect or with a slight decrease in transcription initiation. Thus, the TrC is competent to direct transcription even though it does not do so in nature, and the partial sequence identity it shares with the 3' end of the genome likely represents the important elements of a conserved promoter active in both replication and transcription. Increasing the length of the introduced TrC sequence incrementally to 147 nt resulted in a fourfold increase in replication and a nearly complete inhibition of transcription. These two effects were unrelated, implying that transcription and replication are not interconvertible processes mediated by a common polymerase, but rather are independent processes. The increase in replication was specific to the TrC sequence, implying the presence of a nonessential, replication-enhancing cis-acting element. In contrast, the inhibitory effect on transcription was due solely to the altered spacing between the 3' end of the genome and GS signal, which implies that the transcriptase recognizes the first GS signal as a promoter element. Neither the enhancement of replication nor the inhibition of transcription was due to increased base-pairing potential between the 3' and 5' ends. The relative strengths of the Le and TrC promoters for directing RNA synthesis were compared and found to be very similar. Thus, these findings highlighted a high degree of functional similarity between the RSV antigenomic and genomic promoters, but provided a further distinction between promoter requirements for transcription and replication. PMID- 10846081 TI - Enhancement of primary and secondary cellular immune responses against human immunodeficiency virus type 1 gag by using DNA expression vectors that target Gag antigen to the secretory pathway. AB - In this study, we have investigated the influence of antigen targeting after DNA vaccination upon the induction of cellular immune responses against human immunodeficiency virus type 1 (HIV-1) Gag. In addition to the standard version of HIV-1 Gag, we constructed Gag expression vectors that encode a secreted (Sc-Gag) and a cytoplasmic (Cy-Gag) Gag molecule. Although all three HIV-1 Gag expression vectors induced detectable humoral and cellular immune responses, after intramuscular injection the DNA vector encoding the Sc-Gag generated the highest primary cytotoxic T-lymphocyte (CTL) and T-helper responses. Mice immunized with one of the HIV-1 Gag DNA vectors (but not with the control vector pcDNA3. 1) developed a protective immune response against infection with recombinant vaccinia virus expressing HIV-1 Gag, and this response persisted for 125 days. The magnitude of the protection correlated with the levels of Gag-specific ex vivo CTL activity and the number of CD8(+) T cells producing gamma interferon. The DNA vector encoding the Sc-Gag induced higher levels of protection and greater secondary CTL responses than did the DNA vector encoding Cy-Gag. PMID- 10846083 TI - Balanced hemagglutinin and neuraminidase activities are critical for efficient replication of influenza A virus. AB - The SD0 mutant of influenza virus A/WSN/33 (WSN), characterized by a 24-amino acid deletion in the neuraminidase (NA) stalk, does not grow in embryonated chicken eggs because of defective NA function. Continuous passage of SD0 in eggs yielded 10 independent clones that replicated efficiently. Characterization of these egg-adapted viruses showed that five of the viruses contained insertions in the NA gene from the PB1, PB2, or NP gene, in the region linking the transmembrane and catalytic head domains, demonstrating that recombination of influenza viral RNA segments occurs relatively frequently. The other five viruses did not contain insertions in this region but displayed decreased binding affinity toward sialylglycoconjugates, compared with the binding properties of the parental virus. Sequence analysis of one of the latter viruses revealed mutations in the hemagglutinin (HA) gene, at sites in close proximity to the sialic acid receptor-binding pocket. These mutations appear to compensate for reduced NA function due to stalk deletions. Thus, balanced HA-NA functions are necessary for efficient influenza virus replication. PMID- 10846085 TI - Proteasome-mediated degradation of the papillomavirus E2-TA protein is regulated by phosphorylation and can modulate viral genome copy number. AB - The bovine papillomavirus E2 proteins regulate viral transcription, replication, and episomal genome maintenance. We have previously mapped the major phosphorylation sites of the E2 proteins to serine residues 298 and 301 and shown that mutation of serine residue 301 to alanine leads to a dramatic (10- to 20 fold) increase in viral DNA copy number. In this study we analyzed how phosphorylation regulates E2 protein function. S301 is located in a PEST sequence; these sequences are often found in proteins with a short half-life and can be regulated by phosphorylation. We show here that the E2 protein is ubiquitinated and degraded by the proteasome. Mutation of serine 301 to alanine increases the half-life of E2 from approximately 50 min to 160 min. Furthermore, the A301 E2 protein shows greatly reduced ubiquitination and degradation by the proteasome. These results suggest that the E2 protein level is regulated by phosphorylation, which in turn determines viral episomal copy number. PMID- 10846084 TI - Interactions between brain endothelial cells and human T-cell leukemia virus type 1-infected lymphocytes: mechanisms of viral entry into the central nervous system. AB - Human T-cell leukemia virus type 1 (HTLV-1) is associated with a variety of clinical manifestations, including tropical spastic paraparesis or HTLV-1 associated myelopathy (TSP/HAM). Viral detection in the central nervous system (CNS) of TSP/HAM patients demonstrates the ability of HTLV-1 to cross the blood brain barrier (BBB). To investigate viral entry into the CNS, rat brain capillary endothelial cells were exposed to human lymphocytes chronically infected by HTLV 1 (MT2), to lymphocytes isolated from a seropositive patient, or to a control lymphoblastoid cell line (CEM). An enhanced adhesion to and migration through brain endothelial cells in vitro was observed with HTLV-1-infected lymphocytes. HTLV-1-infected lymphocytes also induced a twofold increase in the paracellular permeability of the endothelial monolayer. These effects were associated with an increased production of tumor necrosis factor alpha by HTLV-1-infected lymphocytes in the presence of brain endothelial cells. Ultrastructural analysis showed that contact between endothelial cells and HTLV-1-infected lymphocytes resulted in a massive and rapid budding of virions from lymphocytes, followed by their internalization into vesicles by brain endothelial cells and apparent release onto the basolateral side, suggesting that viral particles may cross the BBB using the transcytotic pathway. Our study also demonstrates that cell-cell fusion occurs between HTLV-1-infected lymphocytes and brain endothelial cells, with the latter being susceptible to transient HTLV-1 infection. These aspects may help us to understand the pathogenic mechanisms associated with neurological diseases induced by HTLV-1 infection. PMID- 10846086 TI - Binding of Tat to TAR and recruitment of positive transcription elongation factor b occur independently in bovine immunodeficiency virus. AB - Transcriptional transactivators (Tat) from many lentiviruses interact with their cognate transactivation response RNA structures (TAR) to increase rates of elongation rather than initiation of transcription. For several of them, the complex of Tat and a species-specific cyclin T1 must be formed before the binding to TAR can occur with high affinity and specificity. In sharp contrast, Tat from the bovine immunodeficiency virus (BIV) binds to its TAR without the help of the cyclin T1. This binding depends on the upper stem and 5' bulge, but not the central loop in TAR. Moreover, cyclins T1 from different species can mediate effects of this Tat in cells. Unlike the situation with other lentiviruses, Tat transactivation can be rescued simply by linking a heterologous promoter to TAR in permissive cells. Thus, lentiviruses have evolved different strategies to recruit Tat and the positive transcription elongation factor b to their promoters, and interactions between Tat and TAR are independent from those between Tat and the cyclin T1 in BIV. PMID- 10846087 TI - Exacerbation of autoantibody-mediated hemolytic anemia by viral infection. AB - Strong enhancement of the pathogenicity of an antierythrocyte monoclonal antibody was observed after infection of mice with lactate dehydrogenase-elevating virus. While injection of the antierythrocyte antibody alone induced only moderate anemia, concomitant infection with this virus, which is harmless in most normal mice, led to a dramatic drop in the hematocrit and to death of infected animals. In vitro and in vivo analyses showed a dramatic increase in the ability of macrophages from infected mice to phagocytose antibody-coated erythrocytes. These results indicate that viruses can trigger the onset of autoimmune disease by enhancing the pathogenicity of autoantibodies. They may explain how unrelated viruses could be implicated in the etiology of autoantibody-mediated autoimmune diseases. PMID- 10846088 TI - Retargeting the coxsackievirus and adenovirus receptor to the apical surface of polarized epithelial cells reveals the glycocalyx as a barrier to adenovirus mediated gene transfer. AB - Lumenal delivery of adenovirus vectors (AdV) results in inefficient gene transfer to human airway epithelium. The human coxsackievirus and adenovirus receptor (hCAR) was detected by immunofluorescence selectively at the basolateral surfaces of freshly excised human airway epithelial cells, suggesting that the absence of apical hCAR constitutes a barrier to adenovirus-mediated gene delivery in vivo. In transfected polarized Madin-Darby canine kidney cells, wild-type hCAR was expressed selectively at the basolateral membrane, whereas hCAR lacking the transmembrane and/or cytoplasmic domains was expressed on both the basolateral and apical membranes. Cells expressing apical hCAR still were not efficiently transduced by AdV applied to the apical surface. However, after the cells were treated with agents that remove components of the apical surface glycocalyx, AdV transduction occurred. These results indicate that adenovirus can infect via receptors located at the apical cell membrane but that the glycocalyx impedes interaction of AdV with apical receptors. PMID- 10846089 TI - A carboxy-terminally truncated form of the human immunodeficiency virus type 1 Vpr protein induces apoptosis via G(1) cell cycle arrest. AB - Viral protein R (Vpr) of human immunodeficiency virus type 1 inhibits cell proliferation by arresting the cell cycle at the G(2) phase and inducing to apoptosis after G(2) arrest. We have reported previously that C81, a carboxy terminally truncated form of Vpr, interferes with cell proliferation via a novel pathway that is distinct from G(2) arrest. However, the mechanism of this effect of C81 is unknown. We demonstrate here that C81 can induce apoptosis via G(1) arrest of the cell cycle. Immunostaining for various markers of stages of the cell cycle and flow cytometry analysis of DNA content showed that most HeLa cells that had been transiently transfected with a C81 expression vector were arrested at the G(1) phase and not at the G(2) or S phase of the cell cycle. Staining for annexin V, which binds phosphatidylserine on the plasma membrane, as an early indicator of apoptosis and measurement of the activity of caspase-3, a signaling molecule in apoptotic pathways, indicated that C81 is a strong inducer of apoptosis. Expression of C81 induced the condensation, fragmentation, and clumping of chromatin that are typical of apoptosis. Furthermore, the kinetics of the C81-induced G(1) arrest were closely correlated with changes in the number of annexin V-positive cells and the activity of caspase-3. Replacement of Ile or Leu residues by Pro at positions 60, 67, 74, and 81 within the leucine zipper-like domain of C81 revealed that Ile60, Leu67, and Ile74 play important roles both in the C81-induced G(1) arrest and in apoptosis. Thus, it appears that C81 induces apoptosis through pathways that are identical to those utilized for G(1) arrest of the cell cycle. It has been reported that Ile60, Leu67, and Ile74 also play an important role in the C81-induced suppression of growth. These results suggest that the suppression of growth induced by C81 result in apoptosis that is independent of G(2) arrest of the cell cycle. PMID- 10846090 TI - Epstein-Barr virus EB2 protein exports unspliced RNA via a Crm-1-independent pathway. AB - Human herpesviruses encode posttranscriptional activators that are believed to up regulate viral replication by facilitating early and late gene expression. We have reported previously that the Epstein-Barr virus protein EB2 (also called M or SM) promotes nuclear export of RNAs that are poor substrates for spliceosome assembly, an effect that closely resembles the human immunodeficiency virus type 1 Rev-dependent nuclear export of unspliced viral RNA. Here we present experimental data showing that EB2 efficiently promotes the nuclear export of unspliced RNA expressed from a Rev reporter construct. Site-directed mutagenesis as well as domain swapping experiments indicate that a leucine-rich region found in the EB2 protein, which matches the consensus sequence for the leucine-rich nuclear export signal, is not a nuclear export signal per se. Accordingly, leptomycin B (LMB), a specific Crm-1 inhibitor, impairs Rev- but not EB2 dependent nuclear export of unspliced RNA. Moreover, EB2 nucleocytoplasmic shuttling visualized by a heterokaryon assay is, unlike Rev shuttling, not affected by LMB. We also show that overexpression of an N-terminal deletion mutant of Nup214/can, a major nucleoporin of the nuclear pore complex involved in several aspects of nuclear transport, blocks both Rev- and EB2-dependent nuclear export of RNA. These results strongly suggest that EB2 nuclear export of unspliced RNA is mediated by a Crm-1-independent pathway. PMID- 10846091 TI - Gene gun-mediated DNA immunization primes development of mucosal immunity against bovine herpesvirus 1 in cattle. AB - Vaccination by a mucosal route is an excellent approach to the control of mucosally acquired infections. Several reports on rodents suggest that DNA vaccines can be used to achieve mucosal immunity when applied to mucosal tissues. However, with the exception of one study with pigs and another with horses, there is no information on mucosal DNA immunization of the natural host. In this study, the potential of inducing mucosal immunity in cattle by immunization with a DNA vaccine was demonstrated. Cattle were immunized with a plasmid encoding bovine herpesvirus 1 (BHV-1) glycoprotein B, which was delivered with a gene gun either intradermally or intravulvomucosally. Intravulvomucosal DNA immunization induced strong cellular immune responses and primed humoral immune responses. This was evident after BHV-1 challenge when high levels of both immunoglobulin G (IgG) and IgA were detected. Intradermal delivery resulted in lower levels of immunity than mucosal immunization. To determine whether the differences between the immune responses induced by intravulvomucosal and intradermal immunizations might be due to the efficacy of antigen presentation, the distributions of antigen and Langerhans cells in the skin and mucosa were compared. After intravulvomucosal delivery, antigen was expressed early and throughout the mucosa, but after intradermal administration, antigen expression occurred later and superficially in the skin. Furthermore, Langerhans cells were widely distributed in the mucosal epithelium but found primarily in the basal layers of the epidermis of the skin. Collectively, these observations may account for the stronger immune response induced by mucosal administration. PMID- 10846092 TI - Simian immunodeficiency virus rapidly penetrates the cervicovaginal mucosa after intravaginal inoculation and infects intraepithelial dendritic cells. AB - Despite recent insights into mucosal human immunodeficiency virus (HIV) transmission, the route used by primate lentiviruses to traverse the stratified squamous epithelium of mucosal surfaces remains undefined. To determine if dendritic cells (DC) are used by primate lentiviruses to traverse the epithelial barrier of the genital tract, rhesus macaques were intravaginally exposed to cell free simian immunodeficiency virus SIVmac251. We examined formalin-fixed tissues and HLA-DR(+)-enriched cell suspensions to identify the cells containing SIV RNA in the genital tract and draining lymph nodes within the first 24 h of infection. Using SIV-specific fluorescent in situ hybridization combined with immunofluorescent antibody labeling of lineage-specific cell markers, numerous SIV RNA(+) DC were documented in cell suspensions from the vaginal epithelium 18 h after vaginal inoculation. In addition, we determined the minimum time that the SIV inoculum must remain in contact with the genital mucosa for the virus to move from the vaginal lumen into the mucosa. We now show that SIV enters the vaginal mucosa within 60 min of intravaginal exposure, infecting primarily intraepithelial DC and that SIV-infected cells are located in draining lymph nodes within 18 h of intravaginal SIV exposure. The speed with which primate lentiviruses penetrate mucosal surfaces, infect DC, and disseminate to draining lymph nodes poses a serious challenge to HIV vaccine development. PMID- 10846093 TI - Expression of human herpesvirus 6B rep within infected cells and binding of its gene product to the TATA-binding protein in vitro and in vivo. AB - We have characterized the human herpesvirus 6B (HHV-6B) rep gene, which is a homologue of the adeno-associated virus type 2 rep and is unique in the herpesvirus family. Three transcripts, 9.0, 5.0, and 2. 7 kb (the major transcript), were detected by Northern blotting using an HHV-6B rep probe under late conditions. We investigated the expression kinetics of the rep gene using cycloheximide (CHX) and phosphonoformic acid (PFA), which are inhibitors of protein synthesis and viral DNA synthesis, respectively. The 5.2-kb transcript was mainly detected in the absence of protein biosynthesis upon infection, and none of the 9.0-, 5.0-, and 2.7-kb transcripts detected under the late conditions were detected in the presence of CHX and PFA. Sequences obtained from a cDNA library showed that the 5.0- and 2.7-kb transcripts were spliced from two and three exons, respectively, and the 2.7-kb transcript was more abundant. Immunohistochemistry using an antibody raised against the HHV-6 rep gene product (REP) revealed that REP was mainly present in the nucleus of MT-4 cells within 24 h after infection with HHV-6B. Using pull-down assays, coimmunoprecipitation, and a mammalian two hybrid system, we showed that HHV-6 REP binds to a transcription factor, human TATA-binding protein, through its N-terminal region. PMID- 10846094 TI - Depletion of lymphocytes and diminished cytokine production in mice infected with a highly virulent influenza A (H5N1) virus isolated from humans. AB - Previously, we observed that several virulent influenza A (H5N1) viruses which caused severe or fatal disease in humans were also lethal in BALB/c mice following dissemination of the virus to solid organs, including the brain. In contrast, one particular human H5N1 virus was nonlethal in mice and showed no evidence of systemic spread. To compare H5N1 viruses of varying pathogenicity for their ability to alter the mammalian immune system, mice were infected with either influenza A/Hong Kong/483/97 (HK/483) (lethal) or A/Hong Kong/486/97 (HK/486) (nonlethal) virus and monitored for lymphocyte depletion in the blood, lungs, and lymphoid tissue. Intranasal infection with HK/483 resulted in a significant decrease in the total number of circulating leukocytes evident as early as day 2 postinfection. Differential blood counts demonstrated up to an 80% drop in lymphocytes by day 4 postinfection. In contrast, nonlethal HK/486 infected mice displayed only a transient drop of lymphocytes during the infectious period. Analysis of lung and lymphoid tissue from HK/483-infected mice demonstrated a reduction in the number of CD4(+) and CD8(+) T cells and reduced synthesis of the cytokines interleukin-1beta and gamma interferon and the chemokine macrophage inflammatory protein compared with HK/486-infected mice. In contrast, the cytokine and chemokine levels were increased in the brains of mice infected with HK/483 but not HK/486. Evidence of apoptosis in the spleen and lung of HK/483-infected mice was detected in situ, suggesting a mechanism for lymphocyte destruction. These results suggest that destructive effects on the immune system may be one factor that contributes to the pathogenesis of H5N1 viruses in mammalian hosts. PMID- 10846095 TI - The role of CD8(+) T cells and major histocompatibility complex class I expression in the central nervous system of mice infected with neurovirulent Sindbis virus. AB - Little is known about the role of CD8(+) T cells infiltrating the neural parenchyma during encephalitis induced by neurovirulent Sindbis virus (NSV). NSV preferentially infects neurons in the mouse brain and spinal cord; however, it is generally accepted that neurons can express few if any major histocompatibility complex (MHC) class I molecules. We evaluated the possible roles and interactions of CD8(+) T cells during NSV encephalitis and demonstrated that MHC class I antigen (H2K/D) was expressed on endothelial cells, inflammatory cells, and ependymal cells after intracerebral inoculation of NSV. No immunoreactivity was observed in neurons. On the other hand, in situ hybridization with probes for MHC class I heavy chain, beta2 microglobulin, and TAP1 and TAP2 mRNAs revealed increased expression in a majority of neurons, as well as in inflammatory cells, endothelial cells, and ependymal cells in the central nervous system of infected mice. NSV-infected neurons may fail to express MHC class I molecules due to a posttranscriptional block or may express only nonclassical MHC class I genes. To better understand the role CD8(+) T cells play during fatal encephalitis induced by NSV, mice lacking functional CD8(+) T cells were studied. The presence or absence of CD8 did not alter outcome, but absence of beta2 microglobulin improved survival. Interestingly, the intracellular levels of viral RNA decreased more rapidly in immunocompetent mice than in mice without functional CD8(+) T cells. These observations suggest that CD8(+) T cells may act indirectly, possibly via cytokines, to contribute to the clearance of viral RNA in neurons. PMID- 10846096 TI - Identification of the lymantria dispar nucleopolyhedrovirus envelope fusion protein provides evidence for a phylogenetic division of the Baculoviridae. AB - The complete genome sequences of a number of diverse members of the Baculoviridae including both nucleopolyhedroviruses (NPVs) and granuloviruses (GVs) revealed that they lack a homolog of GP64, the envelope fusion protein of the budded form of Autographa californica multinucleocapsid NPV (AcMNPV) and its close relatives. Computer-assisted analyses of the genome of one of these viruses, Lymantria dispar MNPV (LdMNPV), revealed a single open reading frame (ld130) whose product had the predicted properties of a membrane protein. Characterization of the localization of the products of the full-length ld130 gene and of an ld130 enhanced green fluorescent protein gene (egfp) fusion using both immunofluorescence and fluorescence microscopy revealed that LD130 accumulates at the plasma membranes of cells infected with LdMNPV or transfected with ld130 egfp. In addition, cells transfected with either ld130 or ld130-egfp or infected with wild-type virus undergo membrane fusion at pH 5. Western blot analyses indicate that LD130 is present in infected cells as an 83-kDa protein and is also present in budded virions as a protein doublet containing bands of 81 and 83 kDa. Tunicamycin treatment of infected cells resulted in an immunoreactive band of about 72 kDa, indicating that LD130 is N-glycosylated. Whereas the distribution of gp64 appears to be confined to a relatively closely related group of NPVs, homologs of ld130 are present in a diverse number of both NPVs and GVs. This suggests that LD130 may be the primordial baculovirus envelope fusion protein. PMID- 10846097 TI - Dense bodies of human cytomegalovirus induce both humoral and cellular immune responses in the absence of viral gene expression. AB - Infection of fibroblast cell cultures with human cytomegalovirus (HCMV) leads to the production of significant amounts of defective enveloped particles, termed dense bodies (DB). These noninfectious structures contain major antigenic determinants which are responsible for induction of both the humoral and the cellular immune response against HCMV. We tested the hypothesis that, by virtue of their unique antigenic and structural properties, DB could induce a significant immune response in the absence of infectious virus. Mice were immunized with gradient-purified DB, which were either left untreated or subjected to sequential rounds of sonication and freeze-thawing to prevent cellular entry. Titers of neutralizing antibodies induced by DB were in a range comparable to levels present in convalescent human sera. The virus-neutralizing antibody response was surprisingly durable, with neutralizing antibodies detected 12 months following primary immunization. The HCMV-specific major histocompatibility complex class I-restricted cytolytic T-cell (CTL) response was assayed using mice transgenic for the human HLA-A2 molecule. Immunization with DB led to high levels of HCMV-specific CTL in the absence of de novo viral protein synthesis. Maximal total cytolytic activity in mice immunized with DB was nearly as efficient as the cytolytic activity induced by a standard immunization with murine cytomegalovirus. Furthermore, DB induced a typical T-helper 1 (Th1) dominated immune response in mice, as determined by cytokine and immunoglobulin G isotype analysis. Induction of humoral and cellular immune responses was achieved without the concomitant use of adjuvant. We thus propose that DB can serve as a basis for the future development of a recombinant nonreplicating vaccine against HCMV. Finally, such particles could be engineered for efficient delivery of antigens from other pathogens to the immune system. PMID- 10846098 TI - Tumor-specific, replication-competent adenovirus vectors overexpressing the adenovirus death protein. AB - We have constructed two novel adenovirus (Ad) replication-competent vectors, named KD1 and KD3, that may have use in anticancer therapy. The vectors have two key features. First, they markedly overexpress the Ad death protein (ADP), an Ad nuclear membrane glycoprotein required at late stages of infection for efficient cell lysis and release of Ad from cells. Overexpression of ADP was achieved by deleting the E3 region and reinserting the adp gene. Because ADP is overexpressed, KD1 and KD3 are expected to spread more rapidly and effectively through tumors. Second, KD1 and KD3 have two E1A mutations (from the mutant dl1101/1107) that prevent efficient replication in nondividing cells but allow replication in dividing cancer cells. These E1A mutations preclude binding of E1A proteins to p300 and pRB. As a result, the virus should not be able to drive cells from G(0) to S phase and therefore should not be able to replicate in normal tissues. We show that KD1 and KD3 do not replicate well in quiescent HEL 299 cells or in primary human bronchial epithelial cells, small airway epithelial cells, or endothelial cells; however, they replicate well in proliferating HEL 299 cells and human A549 lung carcinoma cells. In cultured A549 cells, KD1 and KD3 lyse cells and spread from cell to cell more rapidly than their control virus, dl1101/1107, or wild-type Ad. They are also more efficient than dl1101/1107 or wild-type Ad in complementing the spread from cell to cell of an E1(-) E3(-) replication-defective vector expressing beta-galactosidase. A549 cells form rapidly growing solid tumors when injected into the hind flanks of immunodeficient nude mice; however, when A549 cells were infected with 10(-4) PFU of KD3/cell prior to injection into mice, tumor formation was nearly completely suppressed. When established A549 tumors in nude mice were examined, tumors injected with buffer grew 13.3-fold over 5 weeks, tumors injected with dl1101/1107 grew 8-fold, and tumors injected with KD1 or KD3 grew 2.6-fold. Hep 3B tumors injected with buffer grew 12-fold over 3.5 weeks, whereas tumors injected with KD1 or KD3 grew 4-fold. We conclude that KD1 and KD3 show promise as anticancer therapeutics. PMID- 10846099 TI - Differences between C57BL/6 and BALB/cBy mice in mortality and virus replication after intranasal infection with neuroadapted Sindbis virus. AB - Neuroadapted Sindbis virus (NSV), given intranasally, caused fatal encephalitis in 100% of adult C57BL/6 mice and 0% of BALB/cBy mice. Most C57BL/6 mice developed severe kyphoscoliosis followed by hind-limb paralysis, while BALB/cBy mice did not. In situ hybridization for detecting NSV RNA and immunohistochemistry for detecting NSV antigen indicated that virus delivered by this route infected neurons of the olfactory region and spread caudally without infection of ependymal cells. Virus antigen was more abundant and infectious virus increased more rapidly and reached higher levels in C57BL/6 mice than in BALB/cBy mice. Surprisingly, infectious virus was cleared faster in C57BL/6 mice, and this was associated with more rapid production of neutralizing antibody. However, viral RNA was cleared more slowly in C57BL/6 mice. In both mouse strains, more infectious virus was present in the lumbar spinal cord than in the cervical spinal cord. These data suggest that genetic susceptibility to fatal NSV encephalomyelitis is determined at least in part by the efficiency of viral replication and spread in the central nervous system. The differences identified in this study provide possible phenotypes for mapping genetic loci involved in susceptibility. PMID- 10846100 TI - The C-terminal proline-rich tail of human immunodeficiency virus type 2 Vpx is necessary for nuclear localization of the viral preintegration complex in nondividing cells. AB - Human immunodeficiency virus type 2 (HIV-2), like other lentiviruses, is capable of infecting nondividing T cells and macrophages. The present work shows that in HIV-2-infected cells, Vpx is necessary for efficient nuclear import of the preintegration complex. In agreement with this finding, the subcellular localization of a GFP-Vpx fusion protein was found to be predominantly nuclear. However, deletion of the proline-rich C-terminal 11 residues of Vpx resulted in a shift of the fusion protein to the cytoplasm. Furthermore, the same deletion in the context of the provirus resulted in a decrease in nuclear import of the preintegration complex and attenuated replication in macrophages. PMID- 10846101 TI - Interaction of human immunodeficiency virus type 2 Vpx and invariant chain. AB - Vpx is a virion-associated protein of human immunodeficiency virus type 2 (HIV-2) and simian immunodeficiency viruses. The yeast two-hybrid system was used to identify invariant chain (Ii) as a cellular protein that interacts with HIV-2 Vpx. Vpx-Ii interaction was confirmed in cell-free reactions using bacterially expressed glutathione S-transferase fusion proteins and by coimmunoprecipitation in transfected and infected cells. In chronically infected cells expressing Vpx, Ii levels were markedly decreased, presumably due to enhanced degradation. These findings suggest that Vpx may disrupt major histocompatibility complex class II antigen presentation. PMID- 10846102 TI - Molecular characterization of an avian astrovirus. AB - Astroviruses are known to cause enteric disease in several animal species, including turkeys. However, only human astroviruses have been well characterized at the nucleotide level. Herein we report the nucleotide sequence, genomic organization, and predicted amino acid sequence of a turkey astrovirus isolated from poults with an emerging enteric disease. PMID- 10846104 TI - Recognition by human monoclonal antibodies of free and complexed peptides representing the prefusogenic and fusogenic forms of human immunodeficiency virus type 1 gp41. AB - Human immunodeficiency virus type 1 (HIV-1) entry into target cells appears to be triggered when two heptad repeat regions in the ectodomain of gp41 associate, converting the prefusogenic form of gp41 to a fusogenic form. Peptides from these two heptad repeat regions, designated N51 and C43, form a coiled coil consisting of an alpha-helical trimer of heterodimers which approximates the core of the fusogenic form of gp41. To understand the antigenic structures of gp41 in these two configurations, and to examine the specificity of anti-gp41 antibodies produced by HIV-1-infected individuals, human anti-gp41 monoclonal antibodies (MAbs) were tested for their reactivity against N51, C43, and the complex formed by these peptides. Of 11 MAbs, 7 reacted with the complex but with neither of the parent peptides. These MAbs reacted optimally with the N51-C43 complex prepared at a 1:1 ratio and appeared to recognize the fusogenic form of gp41 in which the two heptad repeat regions are associated to form the coiled coil. The existence of antibodies from HIV-infected humans that exclusively recognize the N51-C43 complex constitutes the first proof that the coiled-coil conformation of gp41 exists in vivo and is immunogenic. Two of the 11 MAbs were specific for the hydrophilic loop region of gp41 and failed to react with either peptide alone or with the peptide complex, while the remaining 2 MAbs reacted with peptide C43. One of these two latter MAbs, 98-6, also reacted well with the equimolar N51-C43 complex, while reactivity with C43 by the other MAb, 2F5, was inhibited by even small amounts of N51, suggesting that the interaction of these peptides occludes or disrupts the epitope recognized by MAb 2F5. MAbs 98-6 and 2F5 are also unusual among the MAbs tested in their ability to neutralize multiple primary HIV isolates, although 2F5 displays more broad and potent activity. The data suggest that anti-gp41 neutralizing activity is associated with specificity for a region in C43 which participates in complex formation with N51. PMID- 10846103 TI - Variability and immunogenicity of caprine arthritis-encephalitis virus surface glycoprotein. AB - The complete surface glycoprotein (SU) nucleotide sequences of three French isolates of caprine arthritis-encephalitis virus (CAEV) were determined and compared with those of previously described isolates: three American isolates and one French isolate. Phylogenetic analyses revealed the existence of four distinct and roughly equidistant evolutionary CAEV subtypes. Four conserved and five variable domains were identified in the SU. The fine specificities of antibodies produced against these domains during natural infection were examined using a pepscan analysis. Nine immunogenic segments were delineated throughout the conserved and variable domains of SU, two of them corresponding to conserved immunodominant epitopes. Antigenic determinants which may be involved in the immunopathogenic process induced by CAEV were identified. These results also provide sensitive and specific antigen peptides for the serological detection and differentiation of CAEV and visna/maedi virus infections. PMID- 10846105 TI - Resistance to Friend murine leukemia virus infection conferred by the Fv-4 gene is recessive but appears dominant from the effect of the immune system. AB - Fv-4 is a mouse gene that dominantly confers resistance to infection with Friend murine leukemia virus (F-MuLV) (S. Suzuki, Jpn. J. Exp. Med. 45:473-478, 1975). However, the resistance caused by Fv-4 is recessive in nude mice, which suggests that immunological effects play important roles in this resistance in vivo (K. Higo, Y. Kubo, Y. Iwatani, T. Ono, M. Maeda, H. Hiai, T. Masuda, K. Kuribayashi, F. Zhang, T. Lamin, A. Adachi, and A. Ishimoto, J. Virol. 71:750-754, 1997). To determine the immunological effect on the resistance in vivo, we infected immunologically immature newborn mice homozygous (Fv-4(r/r)) and heterozygous (Fv 4(r/-)) for Fv-4. Although the Fv-4(r/r) mice showed complete resistance to F MuLV whether infected neonatally or as adolescents, the Fv-4(r/-) mice showed high sensitivity to viral proliferation and disease induction when infected as newborns but complete resistance when infected as adolescent mice. To confirm the immunological effect on the resistance in adolescent mice with the Fv-4(r/r) and Fv-4(r/-) genotypes, we examined the effect of an immunosuppressant drug, FK506, on the resistance. The mice with the Fv-4(r/r) genotype treated with FK506 still showed resistance, but the mice with the Fv-4(r/-) genotype became highly sensitive to F-MuLV infection. Flow cytometric analysis to detect the Fv-4 gene product showed that the Fv-4 gene product was expressed on the cells from newborn and adolescent mice. The Fv-4 gene product was also detected on the cells from the FK506-treated mice as well as on those from untreated mice. However, a quantitative difference in the gene product between the cells with the Fv-4(r/r) and Fv-4(r/-) genotypes was detected by indirect staining for flow cytometry. These results show that the resistance to F-MuLV infection conferred by the Fv-4 gene is originally recessive, but it looks dominant in adolescent mice mainly because of the effect of the immune system. PMID- 10846106 TI - Isolation of human immunodeficiency virus type 1 cores: retention of Vpr in the absence of p6(gag). AB - Mature human immunodeficiency virus type 1 (HIV-1) virions contain a typically cone-shaped core that encases the viral genome. In this study, we established conditions which allowed the efficient isolation of morphologically intact HIV-1 cores from virions. The isolated cores consisted mostly of cones which appeared uniformly capped at both ends but were heterogeneous with respect to the shape of the broad cap as well as the dimensions and angle of the cone. Vpr, a nonstructural virion component implicated in the nuclear import of the viral genome, was recovered in core preparations of HIV-1 and simian immunodeficiency viruses from African green monkeys. Unexpectedly, p6(gag), a structural protein required for the incorporation of Vpr, was absent from HIV-1 core preparations. Taken together, our results indicate that the incorporation of Vpr into the virion core is a conserved feature of primate lentiviruses and that the interactions required for the uptake of Vpr into assembling particles differ from those which confine Vpr within the core. PMID- 10846107 TI - Influenza virus NS1 protein counteracts PKR-mediated inhibition of replication. AB - The availability of an influenza virus NS1 gene knockout virus (delNS1 virus) allowed us to establish the significance of the biological relationship between the influenza virus NS1 protein and double-stranded-RNA-activated protein kinase (PKR) in the life cycle and pathogenicity of influenza virus. Our results show that the lack of functional PKR permits the delNS1 virus to replicate in otherwise nonpermissive hosts, suggesting that the major function of the influenza virus NS1 protein is to counteract or prevent the PKR-mediated antiviral response. PMID- 10846108 TI - Kaposi's sarcoma-associated herpesvirus open reading frame 50/Rta protein activates the entire viral lytic cycle in the HH-B2 primary effusion lymphoma cell line. AB - Rta, the gene product of Kaposi's sarcoma-associated herpesvirus (KSHV) encoded mainly in open reading frame 50 (ORF50), is capable of activating expression of viral lytic cycle genes. What was not demonstrated in previous studies was whether KSHV Rta was competent to initiate the entire viral lytic life cycle including lytic viral DNA replication, late-gene expression with appropriate kinetics, and virus release. In HH-B2, a newly established primary effusion lymphoma (PEL) cell line, KSHV ORF50 behaved as an immediate-early gene and autostimulated its own expression. Expression of late genes, ORF65, and K8.1 induced by KSHV Rta was eliminated by phosphonoacetic acid, an inhibitor of viral DNA polymerase. Transfection of KSHV Rta increased the production of encapsidated DNase-resistant viral DNA from HH-B2 cells. Thus, introduction of an ORF50 expression plasmid is sufficient to drive the lytic cycle to completion in cultured PEL cells. PMID- 10846109 TI - DNA sequence motifs which direct adeno-associated virus site-specific integration in a model system. AB - The DNA sequence motifs which direct adeno-associated virus type 2 site-specific integration are being investigated using a shuttle vector, propagated as a stable episome in cultured cell lines, as the target for integration. Previously, we reported that the minimum episomal targeting elements comprise a 16-bp binding motif (Rep binding site [RBS]) for a viral regulatory protein (Rep) separated by a short DNA spacer from a sequence (terminal resolution site [TRS]) that can serve as a substrate for Rep-mediated nicking activity (R. M. Linden, P. Ward, C. Giraud, E. Winocour, and K. I. Berns, Proc. Natl. Acad. Sci. USA 93:11288-11294, 1996; R. M. Linden, E. Winocour, and K. I. Berns, Proc. Natl. Acad. Sci. USA 93:7966-7972, 1996). We now report that episomal integration depends upon both the sequence and the position of the spacer DNA separating the RBS and TRS motifs. The spacer thus constitutes a third element required for site-specific episomal integration. PMID- 10846110 TI - Coreceptor-dependent inhibition of the cell fusion activity of simian immunodeficiency virus Env proteins. AB - The cytoplasmic tail (R peptide) sequence is able to regulate the fusion activity of the murine leukemia virus (MuLV) envelope (Env) protein. We have previously shown that this sequence exerts a profound inhibitory effect on the fusion activity of simian immunodeficiency virus (SIV)-MuLV chimeric Env proteins which contain the extracellular and transmembrane domains of the SIV Env protein. Recent studies have shown that SIV can utilize several alternative cellular coreceptors for its fusion and entry into the cell. We have investigated the fusion activity of SIV and SIV-MuLV chimeric Env proteins using cells that express different coreceptors. HeLa cells were transfected with plasmid constructs that carry the SIV or SIV-MuLV chimeric Env protein genes and were overlaid with either CEMx174 cells or Ghost Gpr15 cells, which express the Gpr15 coreceptor for SIV, or Ghost CCR5 cells, which express CCR5, an alternate coreceptor for SIV. The R-peptide sequence in the SIV-MuLV chimeric proteins was found to inhibit the fusion with CEMx174 cells or Ghost Gpr15 cells. However, a significant level of fusion was still observed when HeLa cells expressing the chimeric Env proteins were cocultivated with Ghost CCR5 cells. These results show that the R-peptide sequence exerts differential effects on the fusion activity of SIV Env proteins using target cells that express alternative coreceptors. PMID- 10846111 TI - Genetic analysis of a poliovirus/hepatitis C virus (HCV) chimera: interaction between the poliovirus cloverleaf and a sequence in the HCV 5' nontranslated region results in a replication phenotype. AB - Internal ribosomal entry sites (IRESs) can function in foreign viral genomes or in artificial dicistronic mRNAs. We describe an interaction between the wild-type hepatitis C virus (HCV)-specific sequence and the poliovirus (PV) 5'-terminal cloverleaf in a PV/HCV chimeric virus (containing the HCV IRES), resulting in a replication phenotype. Either a point mutation at nucleotide (nt) 29 or a deletion up to nt 40 in the HCV 5' nontranslated region relieved the replication block, yielding PV/HCV variants replicating to high titers. Fortuitous yet crippling interactions between an IRES and surrounding heterologous RNA must be considered when IRES-based dicistronic expression vectors are being constructed. PMID- 10846113 TI - Endodontics today--a turn-of-the-century reflection. PMID- 10846114 TI - Ferrari temporomandibular disorder instrument. PMID- 10846115 TI - In reply: PMID- 10846112 TI - Respiratory syncytial virus G and/or SH glycoproteins modify CC and CXC chemokine mRNA expression in the BALB/c mouse. AB - Chemokine mRNA expression by pulmonary leukocytes following infection of BALB/c mice with two strains of respiratory syncytial virus (RSV) and one strain of parainfluenza virus type 3 (PIV-3) was determined. The results suggest that RSV G and/or SH proteins inhibit early MIP-1alpha, MIP-1beta, MIP-2, MCP-1, and IP-10 mRNA expression. TCA-3 mRNA expression was found to be increased during PIV-3 infection. PMID- 10846116 TI - Schizophrenia and halitosis. PMID- 10846117 TI - Dislocation of the temporomandibular joint. PMID- 10846118 TI - Swelling of the nasolabial area. PMID- 10846119 TI - Use of keloid skin as an autograft for earlobe reconstruction after excision. PMID- 10846120 TI - Loaded hydroxylapatite-coated implants and uncoated titanium-threaded implants in distracted dog alveolar ridges. AB - AIMS: The aim of this study was to determine the response of alveolar bone after it was augmented vertically with distraction osteogenesis, implanted with hydroxylapatite (HA)-coated implants and noncoated titanium-threaded implants, and subsequently loaded for 1 year. METHODS: Eight dogs each had 4 implants placed horizontally into an edentulous mandibular quadrant. After integration, a distraction osteogenesis device was fabricated in the laboratory. An osteotomy was made to allow the crest of the alveolar ridge to be distracted vertically. After 10 mm of vertical distraction, the distraction devices were stabilized with light cured resin. After bone fill of the distraction gap was radiographically confirmed in all dogs at 10 weeks, 2 implants were placed into the ridges. Four dogs had threaded titanium implants placed, and 4 dogs had threaded HA-coated implants placed, with 1 implant in the distracted bone and 1 implant in adjacent nondistracted bone, for both groups. After 4 months for implant integration, bridges were fabricated and secured to the implants with screws. Crestal bone levels were evaluated by radiographs through 1 year of function. Animals were killed after 1 year of loading for histologic evaluation. RESULTS: The vertical ridge augmentation averaged 8.8 +/- 1.0 mm after 10 weeks of healing after distraction. Analysis of variance indicated a significantly greater change from baseline for HA-coated implants and for distracted bone sites. Histologic examination showed that bone had formed between the distracted segments creating an augmented ridge. The average thickness of the labial cortex in the distraction gap was significantly thinner than the lingual cortex in distracted bone or the lingual and labial nondistracted cortical bone. The presence of a dental implant did not significantly affect cortical bone thickness. Serial sections showed that implants remained integrated and functional without soft tissue inflammation. CONCLUSION: Dental implants placed into alveolar ridges augmented with the technique of distraction osteogenesis were functional for the length of this study. PMID- 10846121 TI - Effect of arthrocentesis and sodium hyaluronate injection on nitrite, nitrate, and thiobarbituric acid-reactive substance levels in the synovial fluid. AB - OBJECTIVE: To evaluate the effect of arthrocentesis and sodium hyaluronate (SH) injections on nitrite, nitrate, and thiobarbituric acid-reactive substance (TBA RS) levels in temporomandibular joint internal derangements. STUDY DESIGN: Arthrocentesis was performed on 10 patients, and 15 patients received a supplemental injection of SH after arthrocentesis. All these patients received an SH injection 15 days after the first intervention. The synovial fluid samples were obtained before arthrocentesis on the first appointment and before the SH injection 15 days later. Nitrite and nitrate levels were measured with a highly sensitive and specific chemiluminescence detection method, and the concentration of lipid peroxidation products was assessed by means of the thiobarbituric acid reaction. RESULTS: Symptomatic improvement was seen in both groups. Nitrite, nitrate, and TBA-RS levels only decreased significantly (P <.05) with a supplemental SH injection after arthrocentesis. CONCLUSIONS: Intra-articular injections of SH may reduce nitrite, nitrate, and TBA-RS levels that play a role in the pathogenesis of various temporomandibular disorders. PMID- 10846122 TI - A clinical comparison of pain perception to the Wand and a traditional syringe. AB - OBJECTIVES: This study compared the pain response of a group of 40 volunteers to the Wand with the response to syringe injections. STUDY DESIGN: Of 240 total injections given, 120 were with the Wand system, and 120 were with the traditional aspirating syringe. Three injections were evaluated: injections to the middle superior alveolar (MSA) of the maxillary right first premolar and the maxillary left first premolar; palatal injections of the maxillary right first premolar and the maxillary left first premolar; and inferior alveolar nerve injection (IAN) of both the right and the left sides. Each volunteer received 6 injections, 3 on one side with the Wand and 3 on the opposite side with the syringe (control). All injections were given by the primary investigator without the use of a topical anesthetic. Pain perception levels were recorded with a 4 point visual analog scale: no pain, mild pain, moderate pain, and severe pain. The results were evaluated statistically by using an unpaired t test. RESULTS: The results showed injections with the syringe were statistically more painful than injections with the Wand in 4 of 18 evaluations: MSA of the maxillary left first premolar, female volunteers responses to MSA of the maxillary left first premolar, IAN left side injections, and male responses to IAN left injections (P =.01, P =.05, P =.05 and P =.05, respectively). CONCLUSIONS: The Wand generally seemed to provide less painful injections; however, the mean ratings of pain were mostly mild pain for both injections. Therefore, the clinical significance of the results should be interpreted with caution. The operator technique and tactile skill in syringe injections and site of injection (right or left) could be important factors that were not evaluated in this study. PMID- 10846123 TI - Minor salivary gland secretion in the elderly. AB - OBJECTIVE: Very little information exists on minor salivary gland output (MSGO) in health and disease. We investigated the minor salivary gland output of elderly subjects with different medical backgrounds and the relationship of that output with life practices, demographic variables, complaints of dry mouth, medication usage, functional status, medical treatments, and dental and medical variables. STUDY DESIGN: MSGO was measured with a special moisture-detecting paper on the lip, cheek, and palate, which comprised the total MSGO. A parsimonious linear regression model was developed to determine which of the many measured variables influenced the MSGO. RESULTS: There were significant differences in MSGO between the recruitment sites, with the non-Veteran Affairs (VA) Independent Living participants having significantly lower MSGO than the VA groups. A secretion rate for MSGO based on median values from all subjects of <4.6 microL/min was categorized as being low, whereas a secretion rate for MSGO >/=4.6 microL/min was categorized as high. A regression model was used to examine variables that could be associated with MSGO variability. The results show that being older, a woman, a current smoker, and having a complaint of dry mouth were associated with lower MSGO rates. CONCLUSIONS: An association between subjective complaints of dry mouth and a decrease in MSGO measured objectively was demonstrated in this elderly population. Furthermore, age and sex effects on MSGO were also demonstrated. PMID- 10846124 TI - Salivary gland disease in human immunodeficiency virus-positive women from the WIHS study. Women's Interagency HIV Study. AB - OBJECTIVE: To determine the prevalence of enlargement, tenderness, and absence of saliva on palpation as indicators of salivary gland disease in women who are human immunodeficiency virus (HIV)-positive. STUDY DESIGN: The study subjects are participants in the Women's Interagency HIV Study (WIHS), a multicenter study examining HIV-seropositive women and at-risk HIV-seronegative women. A total of 576 HIV-positive women and 152 HIV-negative women were examined at their baseline oral visit for clinical markers of salivary gland disease. Viral load levels, CD4 counts, and CD8 counts were obtained as part of the related core study. RESULTS: HIV-positive women had higher rates of salivary gland enlargement (4.3%), tenderness (6.9%), and absence of saliva on palpation (26.6%) compared with HIV negative women, who had rates of 1.3%, 4.6%, and 13.2%, respectively. Absence of saliva was significantly different (P =. 001) between the 2 groups. When 2 of the 3 clinical findings were combined, comparisons between the HIV-positive women and HIV-negative women became significant at the P <.05 level for every combination, except for enlargement/tenderness for the submandibular/sublingual gland. For the HIV-positive women, the viral load was significantly related to enlargement (P =.019) and enlargement/absence of saliva on palpation (P =.037) for the parotids and enlargement (P =.046), absence of saliva (P =.043), and enlargement/absence of saliva (P =.022) for the submandibular/sublingual glands. Significant linear trends were found for increasing viral load and enlargement (P =.013) and enlargement/tenderness (P =.024) for the submandibular/sublingual glands. Significance was present for submandibular/sublingual absence of saliva and tenderness/absence of saliva for CD4 and CD8 medians. CONCLUSIONS: Serostatus is related to salivary gland disease as assessed by glandular enlargement, tenderness, and absence of saliva on palpation. Furthermore, our findings indicate that a multidimensional approach to gland assessment may provide a more complete and perhaps more adequate description of glandular involvement with HIV infection. PMID- 10846125 TI - Salivary epidermal growth factor levels decrease in patients receiving radiation therapy to the head and neck. AB - OBJECTIVE: The objective of this study was to assess changes in salivary epidermal growth factor (EGF) in patients receiving radiation therapy to the head and neck and to determine whether salivary EGF levels correlate with the severity of radiation-induced oral mucositis. STUDY DESIGN: Thirteen patients and 18 control subjects were enrolled in the study. Saliva was collected before, during (weekly), and after radiation therapy. Salivary total protein (TP) and EGF concentrations were measured and correlated with the severity of oral mucositis. The variability in normalized EGF (ngEGF/mgTP) values and mucositis scores were analyzed with analysis of covariance, and the adjusted correlation coefficient was calculated. RESULTS: EGF levels decreased (P =.004), whereas TP levels increased over time (P =.039). A strong correlation was seen with decreasing normalized EGF values and more severe mucositis (P =. 0001). CONCLUSION: A strong negative correlation between normalized EGF and mucositis severity suggests a possible role for EGF in the progression of radiation-induced mucosal breakdown. PMID- 10846126 TI - Oral lichenoid lesions after hepatitis B vaccination. AB - The association of mucocutaneous lichen planus and chronic liver disease is widely recognized. The hepatitis B and C viruses have been implicated as being important in this association, although their exact role remains unclear. Recently, lichenoid lesions of the skin after a hepatitis B vaccination have also been reported. In this case, a woman of Southeast Asian origin had lichenoid lesions affecting the oral mucous membranes develop after she was vaccinated against hepatitis B. The lesions appeared 3 weeks after the administration of the third dose of the vaccine and persisted for about 1 year. As the use of the hepatitis B vaccine becomes more widespread, more such cases can be expected to be encountered. PMID- 10846127 TI - Chronic sclerosing sialadenitis of the submandibular and parotid glands: a report of a case and review of the literature. AB - Chronic sclerosing sialadenitis (also known as Kuttner tumor) is a chronic inflammatory condition of the salivary glands, first described by Kuttner in 1896. Clinically, the disease cannot be distinguished from a true neoplasm. The submandibular gland is affected more commonly than any other salivary gland. This report is of a case of widespread swelling of the salivary glands in which histologic features of chronic sclerosing sialadenitis were seen in the submandibular and parotid glands. The etiology, pathogenesis, and differential diagnosis of this disease and the clinical outcome of this case are discussed and presented. PMID- 10846128 TI - The role of radiotherapy for patients with adenoid cystic carcinoma of the salivary gland. AB - OBJECTIVE: Adenoid cystic carcinoma (ACC) is a rare malignant tumor occurring in the salivary gland that has some characteristic features, including slow growth, extremely diffuse invasion, and a high incidence of distant metastasis. The indication of radiotherapy (RT) for ACC is controversial. The aim of this study was to examine the usefulness of RT for patients with ACC. STUDY DESIGN: The study group was composed of 17 patients (18 lesions) with ACC of the salivary gland who underwent RT. They were divided into 3 groups: (1) those who underwent RT alone for a primary tumor, (2) those who underwent RT alone for a recurrent tumor after surgery, and (3) those who underwent postoperative RT because of a histologically positive surgical margin. The clinical course of each patient was examined retrospectively. RESULTS: Of the 11 patients undergoing RT alone for a primary or recurrent tumor, 5 showed complete remission of the tumor, and 3 were free from local recurrence for a long period. Seven patients who underwent postoperative RT seemed to show better local control than did those who did not undergo postoperative RT, although there were no significant differences. CONCLUSIONS: We find RT to be an effective treatment procedure, especially for those who had an inoperable, advanced tumor or who had distant metastasis. Postoperative RT was recommended for those who had a histologically positive surgical margin. PMID- 10846129 TI - Epithelioid hemangioendothelioma of the parotid salivary gland. AB - Epithelioid hemangioendothelioma is a borderline neoplasm characterized by proliferation of endothelial cells with epithelioid morphology. The tumor, in terms of histology and behavior, occupies an intermediate position between hemangioma and conventional angiosarcoma. It is encountered in a wide variety of sites, such as soft tissues and skin, visceral organs, and bone. This article describes a slowly growing, painful tumor that developed in the parotid gland of a 48-year-old white woman and was treated with a superficial parotidectomy. Microscopically, the lesion consisted of short strands, cords, or small clusters of epithelioid vacuolated cells that exhibited strong immunoreactivity for endothelial cell markers (CD 31, CD 34, and factor VIII-related antigen). Previous reports have presented epithelioid hemangioendotheliomas in the head and neck region but, to our knowledge, this is the first reported case in the parotid salivary gland. PMID- 10846130 TI - Prevalence and extent of long oval canals in the apical third. AB - OBJECTIVE: The aim of this study was to investigate the canal diameters in the apical roots of human teeth to determine prevalence and extent of long oval canals. STUDY DESIGN: This investigation was carried out on 180 extracted human teeth, 20 for each tooth group. Each root was horizontally sectioned at 1, 2, 3, 4, and 5 mm from the apex. Canal diameters were measured with a measuring microscope. RESULTS: In 293 (25%) of the 1181 cross sections investigated, a long oval canal (the long canal diameter was at least 2 times the short canal diameter) was identified. In some tooth groups, the percentage of long oval canals exceeded 50%. In most cases, the long diameter decreased apically; that is, the canal tended toward a rounder cross section. A wide range of diameters existed in all canals. CONCLUSION: Long oval canal is common in the apical 5 mm in human teeth. Many long and narrow oval canals would be impossible to instrument completely without perforating or significantly weakening the roots. Care should be taken in cleaning, shaping, and obturating these oval canals. PMID- 10846131 TI - Checkerboard DNA-DNA hybridization analysis of endodontic infections. AB - OBJECTIVE: The purpose of this investigation was to examine the microbiota of infected root canals by using a molecular genetic method. STUDY DESIGN: The presence and levels of 42 bacterial species were determined in 28 root canal samples by using whole genomic DNA probes and checkerboard DNA-DNA hybridization. To confirm the presence of bacterial DNA in clinical samples, a polymerase chain reaction with an ubiquitous bacterial primer was undertaken. RESULTS: The results of the checkerboard DNA-DNA hybridization analysis showed that 22 of the 42 DNA probes tested were reactive with 1 or more samples. The number of bacterial species in the root canal samples ranged from 1 to 17 (mean, 4.7). Seventeen of the 28 root canal samples were positive for at least 1 DNA probe. The most prevalent species found were as follows: Bacteroides forsythus (39. 3% of the cases); Haemophilus aphrophilus (25%); Corynebacterium matruchotii (21.4%); Porphyromonas gingivalis (17.9%); and Treponema denticola (17.9%). CONCLUSIONS: The microbiologic data of the present investigation indicated that molecular genetic methods can provide significant additional knowledge regarding the endodontic microbiota by detecting bacterial species that are difficult or impossible to culture. In addition, our findings support the current concept that endodontic infections are mixed infections of polymicrobial etiology. PMID- 10846132 TI - Effect of segmental Le Fort I osteotomy on maxillary tooth type-related pulpal blood-flow characteristics. AB - Laser Doppler flowmetry (LDF) is a noninvasive method to assess pulpal blood-flow (PBF). Osteotomies may have segment-related losses of pulpal sensibility. OBJECTIVES: To determine the effect of segmental Le Fort I osteotomy on tooth type related PBF values. MATERIALS AND METHODS: In 12 volunteers, maxillary incisors, canines, and premolars were investigated bilaterally by LDF to assess local PBF values before and after surgery. Perfusion units (PU) were in 3 sessions, on the day before surgery and at 4 and 56 days after osteotomy. RESULTS: Measurements before surgery were significantly higher than at 4 days after surgery for the canine (P <.01) and for the overall PBF values (P <.01). At 4 days assessment, PBF values of tooth types adjacent to vertical osteotomy cuts showed a significant decrease for the lateral incisors (P <.05), canines, and first premolars (P <.05), with no significant differences between the preoperative and postoperative values for tooth types not adjacent to vertical osteotomy cuts (P <.05). CONCLUSIONS: Segmental Le Fort I osteotomy induced a short-term and long-term decrease in maxillary PBF values of tooth types adjacent to vertical osteotomy cuts. PMID- 10846133 TI - The validation of 3D spiral CT-based measurements of simulated maxillofacial neoplasms. AB - OBJECTIVE: The purpose of this study was to determine the precision and accuracy of 3-dimensional spiral computed tomography-based linear measurements of neoplasms associated with the mandible. STUDY DESIGN: Four cadaver heads, each with 2 simulated tumors made of clay, containing contrast medium, and positioned medial to the mandible, were examined by means of a subsecond spiral computed tomography unit. The computed tomography data were transferred to a computer workstation and analyzed through use of 3-dimensional reconstructed images. Linear measurements of the length, width, and depth of the simulated tumors were made by 2 observers, twice each. The soft tissues were then removed and the same measurements made by means of calipers. RESULTS: There were no statistically significant differences between the 3-dimensional computed tomography and physical measurements (P >.05). The mean difference was found to be less than 0.4 mm. CONCLUSIONS: Spiral computed tomography imaging allows for precise and accurate 3-dimensional computed tomography-based measurements for neoplastic lesions in the mandible. PMID- 10846134 TI - Effect of interference of metallic objects on interpretation of T1-weighted magnetic resonance images in the maxillofacial region. AB - PURPOSE: Metallic objects produce artifacts on magnetic resonance (MR) images. However, studies on typical dental materials in this area are scant. This study was conducted to describe and measure the magnitude of such artifacts. MATERIALS AND METHODS: Samples of various dental materials (dental gold, amalgam, stainless steel, titanium, silver-palladium, and vitallium) were embedded in bovine muscle and then subjected to T1-weighted MR imaging. The materials were of the typical morphology and composition encountered in routine maxillofacial practice. The MR artifacts were measured and compared with the original specimen measurements. RESULTS: All metallic objects were found to produce artifacts and to interfere with the interpretation of MR images. Artifacts were most pronounced in the central plane of the object. Gold produced the greatest artifact, and amalgam produced the least. CONCLUSION: Because metals commonly used in the maxillofacial region all produce artifacts on MR images, avoidance measures should be used to minimize the effect of these artifacts. PMID- 10846135 TI - Evaluation of tuned-aperture computed tomography depth discrimination for image series acquired variously with linear horizontal, linear vertical, and conical beam projection arrays. AB - OBJECTIVE: To compare depth discrimination by using tuned-aperture computed tomography (TACT) variously with linear horizontal, linear vertical, combined linear horizontal and linear vertical, and conical beam projection arrays. STUDY DESIGN: The first test object was a metallic mesh angled at 30 degrees to the surface of a computed dental radiography size No. 1 x-ray sensor. The second test object was a dry human mandible. The sensor was mounted on an optical bench constructed to permit free and precise geometrical settings for the horizontal and vertical angulations of the x-ray beam. The extent of blurring of horizontal and vertical wires in each TACT reconstructed image slice was observed for each of the tested beam projection arrays. RESULTS: With a linear horizontal beam projection array, it was not possible to determine the depth of structures parallel to the horizontal dimension, such as the mandibular canal. With a linear vertical beam projection, it was not possible to determine the depth of structures parallel to the vertical dimension. A conical array of beam projections was best suited to the task of depth discrimination of objects in all planes. The best second alternative was a combination of linear vertical and linear horizontal projections. CONCLUSIONS: Beam projection geometry is important for the accurate depth discrimination of TACT reconstructed images. A conical beam projection array is ideal. PMID- 10846137 TI - Good publication practice PMID- 10846136 TI - Analysis of sensitivity and specificity of a new digital subtraction system: an in vitro study. AB - OBJECTIVE: The purpose of this study was to compare a new digital subtraction system with conventional radiograph images for the detection of periapical and periodontal bone lesions. STUDY DESIGN: Periapical and periodontal bone lesions were simulated with cortical bone chips of varying sizes placed on a human dry mandible. Radiographic film images were acquired from varying projections and were subsequently digitized, registered, and subtracted. Four clinicians evaluated the subtracted images, and sensitivity and specificity were calculated. RESULTS: The mean sensitivity and specificity of the Diagnostic Subtraction Radiography system for detecting bone lesions of all sizes with varying projection geometry were 87.90% and 85.23%, respectively. The corresponding results for conventional radiograph images were 47.54% and 97.38%. The difference in sensitivity was statistically significant, whereas the difference in specificity was not. CONCLUSIONS: These results indicate that, even when radiographs are taken from disparate projection geometries, the Diagnostic Subtraction Radiography system is capable of excellent discrimination between healthy and disease states in this in vitro model. PMID- 10846138 TI - The COPE report 1999. Guidelines On good publication practice PMID- 10846139 TI - Who needs a gatekeeper? Patients' views of the role of the primary care physician. AB - BACKGROUND: The primary care physician serving as a 'gatekeeper' can make judicious decisions about the appropriate use of medical services, and thereby contribute to containing costs while improving the quality of care. However, in Israel, sick funds competing for members have not adopted this model for fear of endangering their competitive stance. The purpose of this study was to examine, for the first time, the stated preferences and actual behaviour of a national sample of members of the four Israeli sick funds regarding self-referral to specialists, and to identify the characteristics of patients who prefer the gatekeeper model. METHODS: Data were derived from a national telephone survey carried out in 1997. A random representative sample of 1084 of all adult sick fund members were interviewed, with a response rate of 81%. Bivariate analysis was conducted using over all chi-square tests, and multivariate analysis was performed using logistic regression models. RESULTS: A third of all respondents prefer self-referral to a specialist, 40% prefer their family physician to act as gatekeeper and 19% prefer the physician to co-ordinate care but to refer themselves to a specialist. Independent variables predicting preference for the gatekeeper model are: living in the periphery, sick fund membership, low level of education, being male, fair or poor health status, having a permanent family physician and being satisfied with the professional level of the family physician. A significant correlation was found between practising self-referral and preference for self-referral. CONCLUSIONS.: The findings indicate the importance of surveying patients' attitudes as an input in policy formulation. The study identified specific population groups which prefer the gatekeeper model, and explored the advantages of a flexible model of gatekeeping. PMID- 10846140 TI - Do doctors pay attention to the religious beliefs of their patients? A survey amongst Dutch GPs. AB - BACKGROUND: Patients' religious beliefs can offer support at times of illness and disease. Therefore religious beliefs of patients are important in doctor-patient interaction. OBJECTIVE: To assess to what extent GPs pay attention to religious beliefs of patients in their daily work. METHODS: A postal questionnaire was sent to 120 GPs. The questionnaire consisted of five clusters of items with precoded Likert-scale answer categories related to several clinical situations. RESULTS: Response rate was 72% (n = 87). Upon registration in the practice, 16% of the GPs paid attention to the religious beliefs of patients, while in situations concerning end-of-life decisions like terminal illness or requests for euthanasia most GPs pay attention to religious beliefs of patients (79%). In general GPs brought up in Protestant families tend to pay more attention to religious beliefs of patients than GPs with a Catholic background (65% vs 36%; 95% CI 5-51) and Protestant GPs pay more attention to these aspects than Catholic GPs (81% vs 47%; 95% CI 5-63). CONCLUSIONS: Most GPs tend to pay attention to religion when their medical possibilities in patient care come to an end. GPs and trainees might be conscious of these aspects in patient management. Since most GPs are familiar just with Western religions, the increasing number of non-Western religious denominations might have consequences for patient care in general practitioners' work. PMID- 10846141 TI - Demographic characteristics and primary health care utilization patterns of strictly orthodox Jewish and non-Jewish patients. AB - BACKGROUND: The importance of providing health care services that are acceptable to different cultural groups is widely acknowledged. Strictly orthodox Jewish communities have particular health care needs that reflect their religious teaching and beliefs. OBJECTIVE: To describe the demographic characteristics and health care usage patterns of the strictly orthodox Jewish population of Gateshead. METHODS: Registration and claims data were used in combination with encounter data from computerized and manual practice records. Jewish patients were identified and comparisons made between Jewish and non-Jewish populations registered at the same practices. RESULTS: The orthodox Jewish population was predominantly young (69% aged under 20). The birth rate in orthodox Jewish women aged 20-44 was much higher (294 per 1000) than non-Jewish women. Rates of uptake of cervical screening and childhood immunizations were significantly lower in the orthodox Jewish population. Uptake of breast screening and attendance at diabetic clinics did not differ significantly. The average number of consultations and home visits per annum was higher in Jewish than in non-Jewish patients. CONCLUSIONS: The demographic and health care utilization patterns of orthodox Jewish and non-Jewish patients in Gateshead are different. There are implications for the provision of primary care services, particularly with regard to preventative health care. PMID- 10846142 TI - Comparison of patients' preferences and evaluations regarding aspects of general practice care. AB - BACKGROUND: Although patients' views on health care are perceived to be crucial, insight into the different constructs capturing these views remains limited. OBJECTIVE: The aim of this study was to determine the relationship between patients' preferences and their evaluations of general practice care. METHODS: Patients visiting five rural practices in The Netherlands were asked to complete a questionnaire measuring either their evaluations or their preferences on 44 aspects of general practice care. After at least 3 weeks, those patients who had answered the evaluation questionnaire received the questionnaire measuring their preferences, and vice versa. RESULTS: A total of 449 patients answered both questionnaires (response 70%). The longer the period after the consultation, the lower was the mean percentage of all 44 aspects rated as 'good' in the evaluation questionnaire (P = 0.006) and the higher was the mean percentage of all 44 aspects rated as 'very important' in the preference questionnaire (P = 0.046). The Spearman rank order correlation between the ranking of patients' evaluations and patients' preferences was 0.34, a low although significant correlation (P = 0.024), i.e. the two rank orders do not resemble each other very much. CONCLUSIONS: Patients clearly distinguished their preferences from their evaluations of general practice care. Aspects of general practice care, whether important or not, can be evaluated positively or negatively. Patients' preferences and patients' evaluations are, however, both influenced by the length of the time elapsed since the consultation. PMID- 10846144 TI - Frequent attenders in general practice: an attempt to reduce attendance. AB - BACKGROUND: 'Frequent attenders' in general practice are known to include patients with a variety of problems. Most studies of frequent attenders have not assessed the impact of providing GPs with detailed summaries of the clinical records of these patients on consultation rates. Good medical records are associated with good care. If it is not relatively easy or quick for GPs to ascertain which chronic illness or psychosocial problems the patient has from the records, it will be difficult to manage the patient proactively. OBJECTIVE: The aim of this study was to investigate the impact on the consultation rate of providing a detailed and accessible summary of patients' problems including physical, social and psychological data based on information already recorded in the patients' records. METHOD: A prospective controlled study was made of frequent attenders at one UK general practice comprising four full-time GPs. A total of 104 frequent attenders were identified by examining the lists of patients attending the surgery and by including the names of patients identified by GPs from memory. The final study groups were patients who consulted a GP 11 times or more in a year. The sample was divided into two groups. In both groups, the clinical notes were summarized for data relating to physical, social and psychological problems. In the intervention group, summaries were displayed prominently in the notes which were marked with a distinctive label. GPs were asked to read and initial the available summaries in these notes. In the control group, summaries were not included in the notes and the notes were not highlighted in any way. The consultation rate of these 104 patients was recorded for 5 months after the availability of the summaries in the notes. RESULTS: Summarizing the notes in the style described in this study failed to reduce the consultation rate of the identified frequent attenders (Kruskal-Wallis H = 2.75, P = 0.1) Furthermore, in the intervention group, patients for whom there was evidence that the summaries had been consulted by their GP (24; 46%) continued to attend as frequently as those whose summaries were not endorsed as consulted (mean attendance 4.8 consultations compared with 4.2 in 5 months). CONCLUSION: Frequent attenders often have multiple problems, but prominently displayed summaries of their history for use by GPs during consultations do not reduce the frequency of consultations. PMID- 10846143 TI - Measuring community-oriented attitudes towards medical practice. AB - BACKGROUND: The measurement of attitude and attitudinal changes regarding community-oriented primary care (COPC) and the community-oriented principles of family medicine from the College of Family Physicians of Canada was a key component of this study involving family medicine residents. The Department of Family and Community Medicine at the Toronto Hospital initiated a new COPC curriculum in July 1997 for its first-year residents which was designed to teach the principles of family medicine which are community oriented. OBJECTIVE: This study was developed to provide an analysis and summary of the attitude and attitudinal changes of residents exposed to the programme and those of two cohort groups who were not exposed. METHODS: A quasi-experimental design was used. A 20 item questionnaire was administered pre- and post-intervention. Qualitative data were also collected from focus group sessions with the residents exposed to the programme. RESULTS: The questionnaire was found to have good reliability, with an alpha coefficient of 0.8. No significant differences were observed between the study and control groups pre- and post-intervention. Within the study group, two items from the questionnaire yielded significant differences (P < 0.05). These items dealt with lack of funding and impracticality issues of applying COPC in medical practice. They were also the prevalent themes generated from the focus group session analysis. CONCLUSIONS: The qualitative data corroborated the findings of the survey. These findings have helped in the evolution of the curriculum. Longitudinal studies to measure attitudes and the practice of COPC and community-oriented principles of family medicine after residency are recommended. PMID- 10846145 TI - Defaulters in general practice: who are they and what can be done about them? AB - BACKGROUND: The study of patients in primary care settings who default on their appointment has been based largely on short-term surveys in individual health centres. OBJECTIVE: As part of a wider research project into the potential of practice computer appointment systems as a data source, we wanted to explore the aggregate pattern of default. METHOD: Comprehensive computer appointment data from nine general practices for 1 or 2 years were analysed to explore the pattern of defaulted appointments for doctors and practice nurses. RESULTS: Around 6.5% of all appointments ended in a default. Default rates were found to be highest amongst young adults and, at a practice level, to be highly correlated with deprivation level. About two-thirds of those who defaulted only did it once during the year. A small core of patients defaulted frequently, but only a quarter of these repeated their behaviour in the following year. CONCLUSIONS: The discussion suggests that strategies based on educating or punishing defaulters in order to change their behaviour may be of limited effectiveness. PMID- 10846146 TI - Adherence to the guidelines of a regional formulary. AB - BACKGROUND: Pharmacotherapeutical guidelines, called formularies, have been developed to facilitate effective, efficient and cost-conscious prescribing. Monitoring adherence to such guidelines may be a reasonable way of assessing prescribing practices. OBJECTIVE: The aim of this study was to assess how strictly the GPs participating in our department's registration network adhere to the guidelines of the regional formulary, and which indications and drugs the GPs used. METHODS: This is a descriptive study, concerning 1000 consecutive prescriptions from each of the 17 participating GPs. The third edition of the Groningen formulary (GFIII), published in 1995, was used. If the drug prescribed was advised in the formulary, we considered it to be global adherence. If the indication was mentioned in the formulary, and the drug prescribed was advised for that indication in the formulary, it was considered to be specific adherence. Both the medications prescribed and the health problems registered by the GPs, but not mentioned in the GFIII, were analysed. RESULTS: The 17 000 prescriptions chosen for analysis formed approximately 25% of all prescriptions written by the GPs in 1 year. The indications for only 24 prescriptions (0. 14%) were missing. Among the 17 GPs, the number of different drugs prescribed varied between 167 and 219 per 1000 prescriptions. The global adherence varied from 76 to 89% among the GPs, and the specific adherence varied from 55 to 71%. Of the 17 000 prescriptions, 11 457 (67%) concerned indications mentioned in the GFIII. Prescriptions for indications not mentioned in the GFIII contained 4353 (78.5%) drugs advised in the formulary. Of the 251 medications mentioned in the GFIII, only 15 (6%) were not prescribed. DISCUSSION: The GPs in our study were neither representative, nor were they chosen at random. Their patient population was comparable in age, sex and insurance status. These findings are an example of what level of adherence is obtainable. The formulary covered approximately two thirds of the indications registered by GPs, and did not contain many unnecessary medications (6%). PMID- 10846147 TI - The development of polypharmacy. A longitudinal study. AB - BACKGROUND: To date, only a few studies have been carried out on the development and progress of polypharmacy in relation to morbidity in general practices in The Netherlands. OBJECTIVE: The aim of this study was to investigate the relationship between an increase in long-term drug use and the incidence and severity of some chronic diseases, particularly in the elderly. METHODS: Data on medication and morbidity of 1544 elderly people were collected for the period 1994-1997 from three family practices in the medication and morbidity Registration Network of Groningen (RNG) in the northern part of The Netherlands. Polypharmacy is defined as the long-term simultaneous use of two or more drugs; long-term is defined as >240 days in a year. We looked for differences in incidences of some chronic diseases in those subgroups of the elderly in whom multiple long-term drug use respectively increased, stayed constant or did not exist. Polypharmacy at the end of the period was predicted using regression analysis. RESULTS: Polypharmacy occurred in 42% of the elderly at the end of 1997, with major polypharmacy (>5 drugs) in only 4%. The average number of drugs used long-term increased from 1.3 to 1.8 in 4 years. Predictors for the increase of polypharmacy were the number of drugs at the start, age, diabetes, coronary ischaemic diseases and use of medication without a clear indication (P < 0.005). The average number of diseases also increased, especially in the elderly who showed the greatest increase in long-term drug use; however, there was no significant difference from the groups with a slow or no increase in drug use. DISCUSSION: Polypharmacy showed a slow increase over 4 years: almost 20% of the elderly developed polypharmacy, i.e. going from no drugs or one drug to two or more drugs. Polypharmacy develops mainly in elderly patients who already use several drugs, who are known to suffer from cardiovascular diseases, diabetes or stomach symptoms, those who often take drugs (especially sedatives/hypnotics) without clear indication and those who develop hypertension or atrial fibrillation over time. PMID- 10846149 TI - Mammography screening for breast cancer in women under 50 years PMID- 10846148 TI - Hospitals do not inform GPs about medication that should be monitored. AB - BACKGROUND: General practitioners are now asked to prescribe drugs that, due to possible risks and side effects, had previously been prescribed almost exclusively at hospital. OBJECTIVE: To assess the quality of hospital letters as the key communication between hospitals and GPs. METHOD: Hospital letters examined using a predetermined protocol. RESULTS: Of 224 patients identified who were taking drugs that required regular monitoring, 173 were commenced in hospital. Fewer than one in five (30; 17%) hospital letters indicated that there was a risk associated with the drug or that it should be routinely monitored. Monitoring frequency was identified on only 14 occasions and the majority of letters (129; 74. 6%) did not state who was to be responsible for ongoing monitoring (either GP or hospital). Information was slow to arrive at the practice and, in 12% of cases, the hospital letter had not arrived within 14 days of commencement of medication. CONCLUSION: The information provided in hospital letters is insufficient to allow GPs to put structures in place to monitor drug therapy. PMID- 10846151 TI - Single amino acid (arginine) restriction: growth and death of cultured HeLa and human diploid fibroblasts. AB - Requirements for arginine are different from leucine for the growth of HeLa cells in monolayer and suspension culture. Cells grow increasingly more slowly as arginine levels fall below millimolar. Most cells died at 10(-5) M in static cultures, but could be sustained in perfused cultures, but at 10(-6) M neither perfusion nor increased volume in static cultures compensated. Cell died within 3 4 days in 10(-6) M in the same manner as those in complete arginine deprivation, i.e. considerably faster than with leucine deprivation. Arginine restriction produced by arginase or arginine decarboxylase addition to culture medium gave similar results. Citrulline substituted for arginine, but ornithine and polyamines did not. Arginine was depleted 3-4 times faster from the medium than other amino acids, <5% being consumed in protein synthesis, and arginine released by protein turnover was less efficiently reutilised than leucine. Deprivation reduced protein and DNA syntheses, greatly extended S-phase and protracted the cell cycle in HeLa cells for more than leucine deprivation. The inability of the cells avoid reinitiation of S-phase resulted in their proliferative impetus driving them into an late cycle (premitotic) death. In contrast, normal human diploid fibroblasts reached quiescence with little delay and survived for >11 days. Arginine deprivation is discussed as a selectively means of tumour cell destruction. PMID- 10846153 TI - Neuronal plasticity: increasing the gain in pain. AB - We describe those sensations that are unpleasant, intense, or distressing as painful. Pain is not homogeneous, however, and comprises three categories: physiological, inflammatory, and neuropathic pain. Multiple mechanisms contribute, each of which is subject to or an expression of neural plasticity-the capacity of neurons to change their function, chemical profile, or structure. Here, we develop a conceptual framework for the contribution of plasticity in primary sensory and dorsal horn neurons to the pathogenesis of pain, identifying distinct forms of plasticity, which we term activation, modulation, and modification, that by increasing gain, elicit pain hypersensitivity. PMID- 10846154 TI - Psychological and neural mechanisms of the affective dimension of pain. AB - The affective dimension of pain is made up of feelings of unpleasantness and emotions associated with future implications, termed secondary affect. Experimental and clinical studies show serial interactions between pain sensation intensity, pain unpleasantness, and secondary affect. These pain dimensions and their interactions relate to a central network of brain structures that processes nociceptive information both in parallel and in series. Spinal pathways to limbic structures and medial thalamic nuclei provide direct inputs to brain areas involved in affect. Another source is from spinal pathways to somatosensory thalamic and cortical areas and then through a cortico-limbic pathway. The latter integrates nociceptive input with contextual information and memory to provide cognitive mediation of pain affect. Both direct and cortico-limbic pathways converge on the same anterior cingulate cortical and subcortical structures whose function may be to establish emotional valence and response priorities. PMID- 10846155 TI - Timing the ancestor of the HIV-1 pandemic strains. AB - HIV-1 sequences were analyzed to estimate the timing of the ancestral sequence of the main group of HIV-1, the strains responsible for the AIDS pandemic. Using parallel supercomputers and assuming a constant rate of evolution, we applied maximum-likelihood phylogenetic methods to unprecedented amounts of data for this calculation. We validated our approach by correctly estimating the timing of two historically documented points. Using a comprehensive full-length envelope sequence alignment, we estimated the date of the last common ancestor of the main group of HIV-1 to be 1931 (1915-41). Analysis of a gag gene alignment, subregions of envelope including additional sequences, and a method that relaxed the assumption of a strict molecular clock also supported these results. PMID- 10846157 TI - Forming supramolecular networks from nanoscale rods in binary, phase-separating mixtures AB - Simulations show that when low-volume fractions of nanoscale rods are immersed in a binary, phase-separating blend, the rods self-assemble into needle-like, percolating networks. The interconnected network arises through the dynamic interplay of phase-separation between the fluids, through preferential adsorption of the minority component onto the mobile rods, and through rod-rod repulsion. Such cooperative effects provide a means of manipulating the motion of nanoscopic objects and directing their association into supramolecular structures. Increasing the rod concentration beyond the effective percolation threshold drives the system to self-assemble into a lamellar morphology, with layers of wetted rods alternating with layers of the majority-component fluid. This approach can potentially yield organic/inorganic composites that are ordered on nanometer scales and exhibit electrical or structural integrity. PMID- 10846156 TI - Kinesin superfamily motor protein KIF17 and mLin-10 in NMDA receptor-containing vesicle transport. AB - Experiments with vesicles containing N-methyl-D-aspartate (NMDA) receptor 2B (NR2B subunit) show that they are transported along microtubules by KIF17, a neuron-specific molecular motor in neuronal dendrites. Selective transport is accomplished by direct interaction of the KIF17 tail with a PDZ domain of mLin-10 (Mint1/X11), which is a constituent of a large protein complex including mLin-2 (CASK), mLin-7 (MALS/Velis), and the NR2B subunit. This interaction, specific for a neurotransmitter receptor critically important for plasticity in the postsynaptic terminal, may be a regulatory point for synaptic plasticity and neuronal morphogenesis. PMID- 10846158 TI - Real-space imaging of two-dimensional antiferromagnetism on the atomic scale AB - A two-dimensional antiferromagnetic structure within a pseudomorphic monolayer film of chemically identical manganese atoms on tungsten(110) was observed with atomic resolution by spin-polarized scanning tunneling microscopy at 16 kelvin. A magnetic superstructure changes the translational symmetry of the surface lattice with respect to the chemical unit cell. It is shown, with the aid of first principles calculations, that as a result of this, spin-polarized tunneling electrons give rise to an image corresponding to the magnetic superstructure and not to the chemical unit cell. These investigations demonstrate a powerful technique for the understanding of complicated magnetic configurations of nanomagnets and thin films engineered from ferromagnetic and antiferromagnetic materials used for magnetoelectronics. PMID- 10846159 TI - A nanoplotter with both parallel and serial writing capabilities AB - The development of an eight-pen nanoplotter capable of doing parallel dip-pen nanolithography (DPN) is reported. Because line width and patterning speed in DPN are independent of contact force, only one of the tips in the parallel writing mode (the "imaging" tip) has a feedback system to monitor tip position and to write the pattern; all other tips reproduce what occurs at the imaging tip in a passive fashion. Proof-of-concept experiments that demonstrate eight-pen parallel writing, ink and rinsing wells, and "molecular corralling" via a nanoplotter generated structure are reported. PMID- 10846160 TI - Electronic structure of mott insulators studied by inelastic X-ray scattering AB - The electronic structure of Mott insulators continues to be a major unsolved problem in physics despite more than 50 years of research. Well-developed momentum-resolved spectroscopies such as photoemission or neutron scattering cannot probe the full Mott gap. High-resolution resonant inelastic x-ray scattering revealed dispersive charge excitations across the Mott gap in a high critical temperature parent cuprate (Ca(2)CuO(2)Cl(2)), shedding light on the anisotropy of the Mott gap. These charge excitations across the Mott gap can be described within the framework of the Hubbard model. PMID- 10846161 TI - Millennial-scale instability of the antarctic ice sheet during the last glaciation AB - Records of ice-rafted detritus (IRD) concentration in deep-sea cores from the southeast Atlantic Ocean reveal millennial-scale pulses of IRD delivery between 20,000 and 74,000 years ago. Prominent IRD layers correlate across the Polar Frontal Zone, suggesting episodes of Antarctic Ice Sheet instability. Carbon isotopes (delta(13)C) of benthic foraminifers, a proxy of deepwater circulation, reveal that South Atlantic IRD events coincided with strong increases in North Atlantic Deep Water (NADW) production and inferred warming (interstadials) in the high-latitude North Atlantic. Sea level rise or increased NADW production associated with strong interstadials may have resulted in destabilization of grounded ice shelves and possible surging in the Weddell Sea region of West Antarctica. PMID- 10846162 TI - Extinct (129)I in halite from a primitive meteorite: evidence for evaporite formation in the early solar system AB - Halite crystals from the Zag H3-6 chondrite contain essentially pure (monoisotopic) xenon-129 ((129)Xe) produced in the early history of the solar system by the decay of short-lived iodine-129 ((129)I) (half-life = 15.7 million years). Correlated release of (129)Xe and (128)Xe, produced artificially from (127)I by neutron irradiation, corresponds to an initial ((129)I/(127)I) ratio of (1.35 +/- 0.05) x 10(-4), close to the most primitive early solar system value. If the (129)Xe was produced by in situ decay, then the halite formed from an aqueous fluid within 2 million years of the oldest known solar system minerals. PMID- 10846163 TI - Atomic structure of PDE4: insights into phosphodiesterase mechanism and specificity. AB - Cyclic nucleotides are second messengers that are essential in vision, muscle contraction, neurotransmission, exocytosis, cell growth, and differentiation. These molecules are degraded by a family of enzymes known as phosphodiesterases, which serve a critical function by regulating the intracellular concentration of cyclic nucleotides. We have determined the three-dimensional structure of the catalytic domain of phosphodiesterase 4B2B to 1.77 angstrom resolution. The active site has been identified and contains a cluster of two metal atoms. The structure suggests the mechanism of action and basis for specificity and will provide a framework for structure-assisted drug design for members of the phosphodiesterase family. PMID- 10846164 TI - Signaling specificity by Frizzled receptors in Drosophila. AB - Wnt-Frizzled (Fz) signaling pathways play recurring important roles during the development and homeostasis of vertebrates and invertebrates. Fz receptors can signal through beta-catenin-dependent and -independent pathways. In Drosophila, Fz and Fz2 are redundant receptors for Wg. In addition, Fz conveys signals through a distinct pathway to organize planar polarization of epithelial structures. We demonstrate that the cytoplasmic sequences of Fz2 and Fz preferentially activate the beta-catenin and planar polarity cascade, respectively. Both receptors activate either pathway, but with different efficiencies. Intrinsic differences in signaling efficiency in closely related receptors might be a general mechanism for generating signaling specificity in vivo. PMID- 10846165 TI - Economic incentives for rain forest conservation across scales. AB - Globally, tropical deforestation releases 20 to 30% of anthropogenic greenhouse gases. Conserving forests could reduce emissions, but the cost-effectiveness of this mechanism for mitigation depends on the associated opportunity costs. We estimated these costs from local, national, and global perspectives using a case study from Madagascar. Conservation generated significant benefits over logging and agriculture locally and globally. Nationally, however, financial benefits from industrial logging were larger than conservation benefits. Such differing economic signals across scales may exacerbate tropical deforestation. The Kyoto Protocol could potentially overcome this obstacle to conservation by creating markets for protection of tropical forests to mitigate climate change. PMID- 10846166 TI - mGluR1 in cerebellar Purkinje cells essential for long-term depression, synapse elimination, and motor coordination. AB - Targeted deletion of metabotropic glutamate receptor-subtype 1 (mGluR1) gene can cause defects in development and function in the cerebellum. We introduced the mGluR1alpha transgene into mGluR1-null mutant [mGluR1 (-/-)] mice with a Purkinje cell (PC)-specific promoter. mGluR1-rescue mice showed normal cerebellar long term depression and regression of multiple climbing fiber innervation, events significantly impaired in mGluR1 (-/-) mice. The impaired motor coordination was rescued by this transgene, in a dose-dependent manner. We propose that mGluR1 in PCs is a key molecule for normal synapse formation, synaptic plasticity, and motor control in the cerebellum. PMID- 10846167 TI - Dissociating the role of the dorsolateral prefrontal and anterior cingulate cortex in cognitive control. AB - Theories of the regulation of cognition suggest a system with two necessary components: one to implement control and another to monitor performance and signal when adjustments in control are needed. Event-related functional magnetic resonance imaging and a task-switching version of the Stroop task were used to examine whether these components of cognitive control have distinct neural bases in the human brain. A double dissociation was found. During task preparation, the left dorsolateral prefrontal cortex (Brodmann's area 9) was more active for color naming than for word reading, consistent with a role in the implementation of control. In contrast, the anterior cingulate cortex (Brodmann's areas 24 and 32) was more active when responding to incongruent stimuli, consistent with a role in performance monitoring. PMID- 10846168 TI - Importin beta mediates nuclear translocation of Smad 3. AB - Smad proteins are intracellular mediators of transforming growth factor-beta (TGF beta) and related cytokines. Although ligand-induced nuclear translocation of Smad proteins is clearly established, the pathway mediating this import is yet to be determined. We previously identified a nuclear localization signal (NLS) in the N-terminal region of Smad 3, the major Smad protein involved in TGF-beta signal transduction. This basic motif (Lys(40-)Lys-Leu-Lys-Lys(44)), conserved among all the pathway-specific Smad proteins, is required for Smad 3 nuclear import in response to ligand. Here we studied the nuclear import pathway of Smad 3 mediated by this NLS. We demonstrate that the isolated Smad 3 MH1 domain displays significant specific binding to importin beta, which is diminished or eliminated by mutations in the NLS. Full-size Smad 3 exhibits weak but specific binding to importin beta, which is enhanced after phosphorylation by the type I TGF-beta receptor. In contrast, no interaction was observed between importin alpha and Smad 3 or its MH1 domain, indicating that nuclear translocation of Smad proteins may occur through direct binding to importin beta. We propose that activation of all of the pathway-specific Smad proteins (Smads 1, 2, 3, 5, 8, and 9) exposes the conserved NLS motif, which then binds directly to importin beta and triggers nuclear translocation. PMID- 10846169 TI - Glutathione depletion in PC12 results in selective inhibition of mitochondrial complex I activity. Implications for Parkinson's disease. AB - Oxidative stress appears to play an important role in degeneration of dopaminergic neurons of the substantia nigra (SN) associated with Parkinson's disease (PD). The SN of early PD patients have dramatically decreased levels of the thiol tripeptide glutathione (GSH). GSH plays multiple roles in the nervous system both as an antioxidant and a redox modulator. We have generated dopaminergic PC12 cell lines in which levels of GSH can be inducibly down regulated via doxycycline induction of antisense messages against both the heavy and light subunits of gamma-glutamyl-cysteine synthetase, the rate-limiting enzyme in glutathione synthesis. Down-regulation of glutamyl-cysteine synthetase results in reduction in mitochondrial GSH levels, increased oxidative stress, and decreased mitochondrial function. Interestingly, decreases in mitochondrial activities in GSH-depleted PC12 cells appears to be because of a selective inhibition of complex I activity as a result of thiol oxidation. These results suggest that the early observed GSH losses in the SN may be directly responsible for the noted decreases in complex I activity and the subsequent mitochondrial dysfunction, which ultimately leads to dopaminergic cell death associated with PD. PMID- 10846170 TI - HDAC1, a histone deacetylase, forms a complex with Hus1 and Rad9, two G2/M checkpoint Rad proteins. AB - HDAC1 is a member of the histone deacetylase family, which plays an important role in modulating the eukaryotic chromatin structure. Numerous studies have demonstrated its involvement in transcription and in tumorigenesis. To better understand the functions and regulation of HDAC1, a yeast two-hybrid screening approach was chosen to identify novel interactions involving HDAC1. Human HDAC1 was found to interact specifically in yeast, mammalian cells, and in vitro with the human Hus1 gene product, whose Schizosaccharomyces pombe homolog has been implicated in G(2)/M checkpoint control. Both HDAC1 and Hus1 proteins localize to the nuclei. Furthermore, HDAC1 and Hus1 were found to exist in a complex with Rad9, a known Hus1-interacting factor. In addition, bioinformatics analysis of the protein sequences of Hus1, Rad1, and Rad9, three checkpoint Rad proteins that form a complex, revealed that they all contain a putative proliferating cell nuclear antigen (PCNA) fold, raising the possibility that these factors may bind to DNA in a PCNA-like ring structure. The results reported in this study strongly suggest a novel pathway involving HDAC1 in G(2)/M checkpoint control through the interaction with a functional Rad complex that may utilize a PCNA-like structure. Therefore, physically and functionally similar apparatus may function during G(2)/M checkpoint and DNA replication. PMID- 10846171 TI - Role of activating region 1 of Escherichia coli FNR protein in transcription activation at class II promoters. AB - FNR is an Escherichia coli transcription factor that activates gene expression in response to anaerobiosis at a large number of promoters by making direct contacts with RNA polymerase. At class II FNR-dependent promoters, where the DNA site for FNR overlaps the -35 element, activating region 1 of FNR is proposed to interact with the C-terminal domain of the RNA polymerase alpha-subunit. Using a model class II FNR-dependent promoter, FF(-41.5), we have performed in vivo and in vitro experiments to investigate the role of this interaction. Our results show that FNR, carrying substitutions in activating region 1, is compromised in its ability to promote open complex formation and thus to activate transcription. Abortive initiation assays were used to assess the contribution of activating region 1 of FNR to open complex formation. A new method for the purification of the FNR protein is also described. PMID- 10846172 TI - Aromatic residues and neighboring Arg414 in the (6R)-5,6,7, 8-tetrahydro-L biopterin binding site of full-length neuronal nitric-oxide synthase are crucial in catalysis and heme reduction with NADPH. AB - Nitric-oxide synthase (NOS) requires the cofactor, (6R)-5,6,7, 8 tetrahydrobiopterin (H4B), for catalytic activity. The crystal structures of NOSs indicate that H4B is surrounded by aromatic residues. We have mutated the conserved aromatic acids, Trp(676), Trp(678), Phe(691), His(692), and Tyr(706), together with the neighboring Arg(414) residue within the H4B binding region of full-length neuronal NOS. The W676L, W678L, and F691L mutants had no NO formation activity and had very low heme reduction rates (<0.02 min(-1)) with NADPH. Thus, it appears that Trp(676), Trp(678), and Phe(691) are important to retain the appropriate active site conformation for H4B/l-Arg binding and/or electron transfer to the heme from NADPH. The mutation of Tyr(706) to Leu and Phe decreased the activity down to 13 and 29%, respectively, of that of the wild type together with a dramatically increased EC(50) value for H4B (30-40-fold of wild type). The Tyr(706) phenol group interacts with the heme propionate and Arg(414) amine via hydrogen bonds. The mutation of Arg(414) to Leu and Glu resulted in the total loss of NO formation activity and of the heme reduction with NADPH. Thus, hydrogen bond networks consisting of the heme carboxylate, Tyr(706), and Arg(414) are crucial in stabilizing the appropriate conformation(s) of the heme active site for H4B/l-Arg binding and/or efficient electron transfer to occur. PMID- 10846173 TI - Inhibition of translocation of nascent apolipoprotein B across the endoplasmic reticulum membrane is associated with selective inhibition of the synthesis of apolipoprotein B. AB - In HepG2 cells, inhibition of apolipoprotein B100 (apoB) translocation across the endoplasmic reticulum by an microsomal triglyceride transfer protein (MTP) inhibitor (CP-10447) in the presence of N-acetyl-leucinyl-norleucinal, a proteasomal inhibitor, results in accumulation of newly synthesized apoB in the translocation channel. Here we demonstrated that such accumulation led to a specific reduction of apoB synthesis. ApoB mRNA levels remained unchanged, but we observed reduced rates of elongation of nascent apoB in puromycin-synchronized cells pretreated with MTP inhibitor. This observation was consistent with a longer half-ribosome transit time for the synthesis of apoB in MTP-inhibited cells. Initiation of translation of apoB mRNA was not impaired by MTP inhibition. Overall, these findings suggest that translocation arrest of apoB in the endoplasmic reticulum channel can exert a selective and negative effect on the synthesis of apoB at the stage of elongation. PMID- 10846174 TI - Disruption of the CED-9.CED-4 complex by EGL-1 is a critical step for programmed cell death in Caenorhabditis elegans. AB - In the nematode Caenorhabditis elegans, the apoptotic machinery is composed of four basic elements: the caspase CED-3, the Apaf-1 homologue CED-4, and the Bcl-2 family members CED-9 and EGL-1. The ced-9(n1950) gain-of-function mutation prevents most, if not all, somatic cell deaths in C. elegans. It encodes a CED-9 protein with a glycine-to-glutamate substitution at position 169, which is located within the highly conserved Bcl-2 homology 1 domain. We performed biochemical analyses with the CED-9G169E protein to gain insight into the mechanism of programmed cell death. We find that CED-9G169E retains the ability to bind both EGL-1 and CED-4, although its affinity for EGL-1 is reduced. In contrast to the behavior of wild-type CED-9, the interaction between CED-9G169E and CED-4 is not disrupted by expression of EGL-1. Furthermore, CED-4 and CED 9G169E co-localizes with EGL-1 to the mitochondria in mammalian cells, and expression of EGL-1 does not induce translocation of CED-4 to the cytosol. Finally, the ability of EGL-1 to promote apoptosis is impaired by the replacement of wild-type CED-9 with CED-9G169E, and this effect is correlated with the inability of EGL-1 to induce the displacement of CED-4 from the CED-9.CED-4 complex. These studies suggest that the release of CED-4 from the CED-9.CED-4 complex is a necessary step for induction of programmed cell death in C. elegans. PMID- 10846175 TI - Insulin receptor substrate-1, p70S6K, and cell size in transformation and differentiation of hemopoietic cells. AB - After an initial burst of cell proliferation, the type 1 insulin-like growth factor receptor (IGF-IR) induces granulocytic differentiation of 32D IGF-IR cells, an interleukin-3-dependent murine hemopoietic cell line devoid of insulin receptor substrate-1 (IRS-1). The combined expression of the IGF-IR and IRS-1 (32D IGF-IR/IRS-1 cells) inhibits IGF-I-mediated differentiation, and causes malignant transformation of 32D cells. Because of the role of IRS-1 in changing the fate of 32D IGF-IR cells from differentiation (and subsequent cell death) to malignant transformation, we have looked for differences in IGF-IR signaling between 32D IGF-IR and 32D IGF-IR/IRS-1 cells. In this report, we have focused on p70(S6K), which is activated by the IRS-1 pathway. We find that the ectopic expression of IRS-1 and the inhibition of differentiation correlated with a sustained activation of p70(S6K) and an increase in cell size. Phosphorylation in vivo of threonine 389 and, to a lesser extent, of threonine 421/serine 424 of p70(S6K) seemed to be a requirement for inhibition of differentiation. A role of IRS-1 and p70(S6K) in the alternative between transformation or differentiation of 32D IGF-IR cells was confirmed by findings that inhibition of p70(S6K) activation or IRS-1 signaling, by rapamycin or okadaic acid, induced differentiation of 32D IGF-IR/IRS-1 cells. We have also found that the expression of myeloperoxidase mRNA (a marker of differentiation, which sharply increases in 32D IGF-IR cells), does not increase in 32D IGF-IR/IRS-1 cells, suggesting that the expression of IRS-1 in 32D IGF-IR cells causes the extinction of the differentiation program initiated by the IGF-IR, while leaving intact its proliferation program. PMID- 10846176 TI - Nitric oxide down-regulates MKP-3 mRNA levels: involvement in endothelial cell protection from apoptosis. AB - MAP kinase-dependent phosphorylation processes have been shown to interfere with the degradation of the antiapoptotic protein Bcl-2. The cytosolic MAP kinase phosphatase MAP kinase phosphatase-3 (MKP-3) induces apoptosis of endothelial cells in response to tumor necrosis factor alpha (TNFalpha) via dephosphorylation of the MAP kinase ERK1/2, leading to Bcl-2 proteolysis. Here we report that the endothelial cell survival factor nitric oxide (NO) down-regulated MKP-3 by destabilization of MKP-3 mRNA. This effect of NO was paralleled by a decrease in MKP-3 protein levels. Moreover, ERK1/2 was found to be protected against TNFalpha induced dephosphorylation by coincubation of endothelial cells with the NO donor. Subsequently, both the decrease in Bcl-2 protein levels and the mitochondrial release of cytochrome c in response to TNFalpha were largely prevented by exogenous NO. In cells overexpressing MKP-3, no differences in phosphatase activity in the presence or absence of NO were found, excluding potential posttranslational modifications of MKP-3 protein by NO. These data demonstrate that upstream of the S-nitrosylation of caspase-3, NO exerts additional antiapoptotic effects in endothelial cells, which rely on the down-regulation of MKP-3 mRNA. PMID- 10846177 TI - Human Cdc7-related kinase complex. In vitro phosphorylation of MCM by concerted actions of Cdks and Cdc7 and that of a criticial threonine residue of Cdc7 bY Cdks. AB - huCdc7 encodes a catalytic subunit for Saccharomyces cerevisae Cdc7-related kinase complex of human. ASK, whose expression is cell cycle-regulated, binds and activates huCdc7 kinase in a cell cycle-dependent manner (Kumagai, H., Sato, N., Yamada, M., Mahony, D. , Seghezzi, W., Lees, E., Arai, K., and Masai, H. (1999) Mol. Cell. Biol. 19, 5083-5095). We have expressed huCdc7 complexed with ASK regulatory subunit using the insect cell expression system. To facilitate purification of the kinase complex, glutathione S-transferase (GST) was fused to huCdc7 and GST-huCdc7-ASK complex was purified. GST-huCdc7 protein is inert as a kinase on its own, and phosphorylation absolutely depends on the presence of the ASK subunit. It autophosphorylates both subunits in vitro and phosphorylates a number of replication proteins to different extents. Among them, MCM2 protein, either in a free form or in a MCM2-4-6-7 complex, serves as an excellent substrate for huCdc7-ASK kinase complex in vitro. MCM4 and MCM6 are also phosphorylated by huCdc7 albeit to less extent. MCM2 and -4 in the MCM2-4-6-7 complex are phosphorylated by Cdks as well, and prior phosphorylation of the MCM2 4-6-7 complex by Cdks facilitates phosphorylation of MCM2 by huCdc7, suggesting collaboration between Cdks and Cdc7 in phosphorylation of MCM for initiation of S phase. huCdc7 and ASK proteins can also be phosphorylated by Cdks in vitro. Among four possible Cdk phosphorylation sites of huCdc7, replacement of Thr-376, corresponding to the activating threonine of Cdk, with alanine (T376A mutant) dramatically reduces kinase activity, indicative of kinase activation by phosphorylation of this residue. In vitro, Cdk2-Cyclin E, Cdk2-Cyclin A, and Cdc2 Cyclin B, but not Cdk4-Cyclin D1, phosphorylates the Thr-376 residue of huCdc7, suggesting possible regulation of huCdc7 by Cdks. PMID- 10846178 TI - Requirement of V-ATPase for ovulation and embryogenesis in Caenorhabditis elegans. AB - Immunofluorescence analysis indicated that VHA-11, the C subunit of Caenorhabditis elegans V-ATPase, was localized in dot-like structures around the nuclei of early embryonic cells and was also detected in embryonic intestinal cells after comma stage. Vital staining with acridine orange showed that the intestinal cells had acidic compartments generated by V-ATPase, consistent with the intracellular localization of VHA-11. RNA interference could efficiently silence vha-11 gene expression: introduction of vha-11 double strand RNA led to embryonic lethality. Worms injected with the vha-11 double strand RNA produced embryos that became lethal. The development of embryos was arrested at various stages. However, their numbers gradually decreased, and the worms eventually became sterile due to the failure of ovulation. Similar results were obtained for RNA interference of the V-ATPase proteolipid genes. These results suggest that V ATPases, and thus inside-acidic organelles, are required for ovulation and embryogenesis. PMID- 10846179 TI - Interaction between transmembrane domains five and six of the alpha -factor receptor. AB - The alpha-factor pheromone receptor (STE2) activates a G protein signal pathway that induces conjugation of the yeast Saccharomyces cerevisiae. Previous studies implicated the third intracellular loop of this receptor in G protein activation. Therefore, the roles of transmembrane domains five and six (TMD5 and -6) that bracket the third intracellular loop were analyzed by scanning mutagenesis in which each residue was substituted with cysteine. Out of 42 mutants examined, four constitutive mutants and two strong loss-of-function mutants were identified. Double mutants combining Cys substitutions in TMD5 and TMD6 gave a broader range of phenotypes. Interestingly, a V223C mutation in TMD5 caused constitutive activity when combined with the L247C, L248C, or S251C mutations in TMD6. Also, the L226C mutation in TMD5 caused constitutive activity when combined with either the M250C or S251C mutations in TMD6. The residues affected by these mutations are predicted to fall on one side of their respective helices, suggesting that they may interact. In support of this, cysteines substituted at position 223 in TMD5 and position 247 in TMD6 formed a disulfide bond, providing the first direct evidence of an interaction between these transmembrane domains in the alpha-factor receptor. Altogether, these results identify an important region of interaction between conserved hydrophobic regions at the base of TMD5 and TMD6 that is required for the proper regulation of receptor signaling. PMID- 10846180 TI - Binding sites of leukocyte beta 2 integrins (LFA-1, Mac-1) on the human ICAM-4/LW blood group protein. AB - The red cell ICAM-4/LW blood group glycoprotein, which belongs to the family of intercellular adhesion molecules (ICAMs), has been reported to interact with CD11a/CD18 (LFA-1) and CD11b/CD18 (Mac-1) beta(2) integrins. To better define the basis of the ICAM-4/beta(2) integrin interaction, we have generated wild-type, domain-deleted and mutated recombinant chimeric ICAM-4-Fc proteins and analyzed their interaction in a cellular adhesion assay with LFA-1 and Mac-1 L-cell stable transfectants. We found that monoclonal antibodies against CD11a, CD11b, CD18, or LW(ab) block adhesion of transfectant L-cells to immobilized ICAM-4-Fc protein and that the ICAM-4/beta(2) integrin interaction was highly sensitive to the presence of the divalent cations Ca(2+) and Mg(2+). Deletion of individual Ig domains D1 or D2 of the extracellular part of ICAM-4 showed that LFA-1 binds to the first Ig-like domain, whereas the Mac-1 binding site encompassed both the first and the second Ig-like domains. Based on the crystal structure of ICAM-2, we propose a model for the Ig-like domains D1 and D2 of ICAM-4. Accordingly, by site-directed mutagenesis of 22 amino acid positions spread out on all faces of the ICAM-4 molecule, we identified four exposed residues, Leu(80), Trp(93), and Arg(97) on the CFG face and Trp(77) on the E-F loop of domain D1 that may contact LFA-1 as part of the binding site. However, the single and double mutants R52E and T91Q on the CFG face of domain D1, which correspond to the key residues Glu(34) and Gln(73) for ICAM-1 binding to LFA-1, had no effect on LFA-1 binding. In contrast, all mutants on the CFG face of domain D1 and residues Glu(151) and Thr(154) in the C'-E loop of the domain D2 seem to play a dominant role in Mac-1 binding. These data suggest that the binding site for LFA-1 on ICAM-4 overlaps but is distinct from the Mac-1 binding site. PMID- 10846182 TI - NMR studies of active N-terminal peptides of stromal cell-derived factor-1. Structural basis for receptor binding. AB - Stromal cell-derived factor 1 (SDF-1), a member of the CXC chemokine family, is the only chemokine to bind to the receptor CXCR4. This receptor is also a co receptor for syncytia-inducing forms of HIV in CD4(+) cells. In addition, SDF-1 is responsible for attracting mature lymphocytes to the bone marrow and can therefore contribute to host versus graft rejection in bone marrow transplantation. Clearly, by manipulating SDF-1 activity, we could find a possible anti-viral AIDS treatment and aid in bone marrow transplantation. SDF-1 binds to CXCR4 primarily via the N terminus, which appears flexible in the recently determined three-dimensional structure of SDF-1. Strikingly, short N terminal SDF-1 peptides have been shown to have significant SDF-1 activity. By using NMR, we have determined the major conformation of the N terminus of SDF-1 in a 17-mer (residues 1-17 of SDF-1) and a 9-mer dimer (residues 1-9 of SDF-1 linked by a disulfide bond at residue 9). Residues 5-8 and 11-14 form similar structures that can be characterized as a beta-turn of the beta-alphaR type. These structural motifs are likely to be interconverting with other states, but the major conformation may be important for recognition in receptor binding. These results suggest for the first time that there may be a link between structuring of short N-terminal chemokine peptides and their ability to activate their receptor. These studies will act as a starting point for synthesizing non peptide analogs that act as CXCR4 antagonists. PMID- 10846181 TI - Correlating a protein structure with function of a bacterial mechanosensitive channel. AB - MscL, a mechanosensitive channel found in many bacteria, protects cells from hypotonic shock by reducing intracellular pressure through release of cytoplasmic osmolytes. First isolated from Escherichia coli, this protein has served as a model for how a protein senses and responds to membrane tension. Recently the structure of a functionally uncharacterized MscL homologue from Mycobacterium tuberculosis was solved by x-ray diffraction to a resolution of 3.5 A. Here we demonstrate that the protein forms a functional MscL-like mechanosensitive channel in E. coli membranes and azolectin proteoliposomes. Furthermore, we show that M. tuberculosis MscL crystals, when re-solubilized and reconstituted, yield wild-type channel currents in patch clamp, demonstrating that the protein does not irreversibly change conformation upon crystallization. Finally, we apply functional clues acquired from the E. coli MscL to the M. tuberculosis channel and show a mechanistic correlation between these channels. However, the inability of the M. tuberculosis channel to gate at physiological membrane tensions, demonstrated by in vivo E. coli expression and in vitro reconstitution, suggests that the membrane environment or other additional factors influence the gating of this channel. PMID- 10846184 TI - Src-mediated tyrosine phosphorylation of NR2 subunits of N-methyl-D-aspartate receptors protects from calpain-mediated truncation of their C-terminal domains. AB - Src-mediated tyrosine phosphorylation of N-methyl-d-aspartate receptor subunits has been shown to modify the functional properties of N-methyl-d-aspartate receptors. Moreover, calpain-mediated truncation of N-methyl-d-aspartate receptor subunits has been found to alter the structure of the receptors. In the present study, we first used immunoprecipitation with a variety of antibodies against N methyl-d-aspartate receptor subunits and anti-phosphotyrosine antibodies to show that tyrosine-phosphorylated subunits of N-methyl-d-aspartate receptor are protected against calpain-mediated truncation of their C-terminal domains. A GST fusion protein containing the C-terminal domain of NR2A was used to identify the calpain cutting sites in the C-terminal domain. One site was identified at residues 1278-1279, corresponding to one of the preferred calpain truncation sites. This site is adjacent to a consensus sequence for Src-mediated tyrosine phosphorylation, and Src-mediated tyrosine phosphorylation of the GST-NR2A C terminal fusion protein also inhibited calpain-mediated truncation of the fusion protein. We propose that phosphorylation of NR2 subunits and the resulting inhibition of calpain-mediated truncation of their C-terminal domains provide for the stabilization of the N-methyl-d-aspartate receptors in postsynaptic structures. PMID- 10846185 TI - Immunological quantitation and localization of ACAT-1 and ACAT-2 in human liver and small intestine. AB - By using specific anti-ACAT-1 antibodies in immunodepletion studies, we previously found that ACAT-1, a 50-kDa protein, plays a major catalytic role in the adult human liver, adrenal glands, macrophages, and kidneys but not in the intestine. Acyl-coenzyme A:cholesterol acyltransferase (ACAT) activity in the intestine may be largely derived from a different ACAT protein. To test this hypothesis, we produced specific polyclonal anti-ACAT-2 antibodies that quantitatively immunodepleted human ACAT-2, a 46-kDa protein expressed in Chinese hamster ovary cells. In hepatocyte-like HepG2 cells, ACAT-1 comprises 85-90% of the total ACAT activity, with the remainder attributed to ACAT-2. In adult intestines, most of the ACAT activity can be immunodepleted by anti-ACAT-2. ACAT 1 and ACAT-2 do not form hetero-oligomeric complexes. In differentiating intestinal enterocyte-like Caco-2 cells, ACAT-2 protein content increases by 5-10 fold in 6 days, whereas ACAT-1 protein content remains relatively constant. In the small intestine, ACAT-2 is concentrated at the apices of the villi, whereas ACAT-1 is uniformly distributed along the villus-crypt axis. In the human liver, ACAT-1 is present in both fetal and adult hepatocytes. In contrast, ACAT-2 is evident in fetal but not adult hepatocytes. Our results collectively suggest that in humans, ACAT-2 performs significant catalytic roles in the fetal liver and in intestinal enterocytes. PMID- 10846186 TI - Epidermal growth factor receptor transactivation mediates substance P-induced mitogenic responses in U-373 MG cells. AB - Ligand-induced activation of G protein-coupled receptors is emerging as an important pathway leading to the activation of certain receptors with intrinsic tyrosine kinase activity, such as the epidermal growth factor receptor (EGFR). Substance P (SP) exerts many effects via activation of its G protein-coupled receptor (neurokinin-1, NK-1). SP participates in acute inflammation and activates key proteins involved in mitogenic pathways, such mitogen-activated protein kinases (MAPKs), stimulating DNA synthesis. We tested the hypothesis that SP-induced MAPK activation and DNA synthesis require activation of the EGFR. In U 373 MG cells, which express functional NK-1, SP induced tyrosine phosphorylation of several proteins including EGFR. SP induced formation of an activated EGFR complex containing the adapter proteins SHC and Grb2, but not c-Src. SP activated the MAPK pathway as shown by increased Erk2 kinase activity. SP induced Erk2 activation, and DNA synthesis was inhibited in cells transfected with a dominant negative EGFR plasmid lacking kinase activity, as well as in cells treated with a specific EGFR inhibitor. In addition, pertussis toxin, an inhibitor of Galpha(iota) protein subunits, prevented SP-induced EGFR transactivation and subsequent DNA synthesis. Our results implicate EGFR as an essential regulator in SP/NK-1-induced activation of the MAPK pathway and cell proliferation in U-373 MG cells, and these events are mediated by a pertussis toxin-sensitive Galpha protein. We suggest that this mechanism by which SP controls cell proliferation is an important pathway in tissue restoration and healing. PMID- 10846188 TI - Work and health: a call for action PMID- 10846187 TI - Mutant presenilin 2 transgenic mice. A large increase in the levels of Abeta 42 is presumably associated with the low density membrane domain that contains decreased levels of glycerophospholipids and sphingomyelin. AB - The N141I mutation in presenilin (PS) 2 is tightly linked with a form of autosomal dominant familial Alzheimer's disease in the Volga German families. We previously reported that mouse brains harboring mutant PS2 contained increased levels of amyloid beta protein (Abeta) 42 in the Tris-saline-soluble fraction (Oyama, F., Sawamura, N., Kobayashi, K., Morishima-Kawashima, M., Kuramochi, T., Ito, M., Tomita, T., Maruyama, K., Saido, T. C., Iwatsubo, T., Capell, A., Walter, J., Grunberg, J., Ueyama, Y., Haass, C. and Ihara, Y. (1998) J. Neurochem. 71, 313-322). Here, using a new extraction protocol, we quantitated the Abeta40 and Abeta42 levels in the Tris-saline-insoluble fraction. The insoluble Abeta levels were found to be higher than the soluble Abeta levels, and the insoluble Abeta42 levels were markedly increased in mutant PS2 transgenic mice. To investigate the origin of the insoluble Abeta42, we prepared the detergent-insoluble, low density membrane fraction. This fraction from two independent lines of mutant PS2 transgenic mice contained remarkably increased levels of Abeta42 and significantly low levels of glycerophospholipids and sphingomyelin. This unexpected finding suggests that a large increase in the levels of Abeta42 in mutant PS2 mice is presumably induced through alterations of the lipid composition in the low density membrane domain in the brain. PMID- 10846189 TI - Working conditions and sickness absence: the need for action oriented research. PMID- 10846190 TI - Effect of change in the psychosocial work environment on sickness absence: a seven year follow up of initially healthy employees. AB - STUDY OBJECTIVE: To investigate the impact of changes in psychosocial work environment on subsequent sickness absence. DESIGN: Analysis of questionnaire and sickness absence data collected in three time periods: 1990-1991, before the recession; 1993, worst slump during the recession; and 1993-1997, a period after changes. SETTING: Raisio, a town in south western Finland, during and after a period of economic decline. PARTICIPANTS: 530 municipal employees (138 men, 392 women) working during 1990-1997 who had no medically certified sick leaves in 1991. Mean length of follow up was 6.7 years. MAIN RESULTS: After adjustment for the pre-recession levels, the changes in the job characteristics of the workers during the recession predicted their subsequent sick leaves. Lowered job control caused a 1.30 (95% CI = 1.19, 1.41) times higher risk of sick leave than an increase in job control. The corresponding figures in relation to decreased social support and increased job demands were 1.30 (95% CI = 1.20, 1.41) and 1.10 (95% CI = 1.03, 1.17), respectively. In some cases there was an interaction with socioeconomic status, changes in the job characteristics being stronger predictors of sick leaves for employees with a high income than for the others. The highest risks of sick leave (ranging from 1.40 to 1.90) were associated with combined effects related to poor levels of and negative changes in job control, job demands and social support. CONCLUSION: Negative changes in psychosocial work environment have adverse effects on the health of employees. Those working in an unfavourable psychosocial environment before changes are at greatest risk. PMID- 10846191 TI - How do types of employment relate to health indicators? Findings from the second European survey on working conditions. AB - STUDY OBJECTIVE: To investigate the associations of various types of employment with six self reported health indicators, taking into account the part played by demographic variables, individual working conditions and four ecological indicators at the country level. DESIGN: Cross sectional survey (structured interview) of a sample of the active population of 15 European countries aged 15 years or over. Main independent variables were nine types of employment categorised as follows: small employers, full and part time permanent employees, full and part time fixed term employees, full and part time sole traders and full and part time temporary contracts. Main outcome measures were three self reported health related outcomes (job satisfaction, health related absenteeism, and stress) and three self reported health problems (overall fatigue, backache, and muscular pains). Logistic regression and multilevel models were used in the analyses. SETTING: 15 countries of the European Union. PARTICIPANTS: 15 146 employed persons aged 15 or over. MAIN RESULTS: Precarious employment was consistently and positively associated with job dissatisfaction but negatively associated with absenteeism and stress (as compared with full time permanent workers). Fatigue, backache and muscular pains also tended to be positively associated with precarious employment, particularly with full time precarious employment. Small employers reported high percentages of stress and fatigue, but absenteeism was relatively low. Sole traders generally reported high percentages of all outcomes, except for absenteeism, which was low. For each type of employment (except temporary contracts), full time workers tended to report worse health outcomes than part time workers. Patterns were generally consistent across countries. Associations persisted after adjustment for individual level working conditions and were not modified by country level variables. CONCLUSIONS: This study is the first to examine the relations between various types of employment and six health related indicators for all 15 member states of the European Union. Suggestive patterns worthy of further exploration have been found. Standardised definitions of types of underemployment and health related outcomes, more potent epidemiological designs and the inclusion of socioeconomic information (for example, social security systems, incapacity benefit schemes) at the regional level are proposed for inclusion in further research. PMID- 10846192 TI - Sick but yet at work. An empirical study of sickness presenteeism. AB - STUDY OBJECTIVE: The study is an empirical investigation of sickness presenteeism in relation to occupation, irreplaceability, ill health, sickness absenteeism, personal income, and slimmed down organisation. DESIGN: Cross sectional design. SETTING: Swedish workforce. PARTICIPANTS: The study group comprised a stratified subsample of 3801 employed persons working at the time of the survey, interviewed by telephone in conjunction with Statistics Sweden's labour market surveys of August and September 1997. The response rate was 87 per cent. MAIN RESULTS: A third of the persons in the total material reported that they had gone to work two or more times during the preceding year despite the feeling that, in the light of their perceived state of health, they should have taken sick leave. The highest presenteeism is largely to be found in the care and welfare and education sectors (nursing and midwifery professionals, registered nurses, nursing home aides, compulsory school teachers and preschool/primary educationalists. All these groups work in sectors that have faced personnel cutbacks during the 1990s). The risk ratio (odds ratio (OR)) for sickness presenteeism in the group that has to re-do work remaining after a period of absence through sickness is 2.29 (95% CI 1.79, 2.93). High proportions of persons with upper back/neck pain and fatigue/slightly depressed are among those with high presenteeism (p< 0.001). Occupational groups with high sickness presenteeism show high sickness absenteeism (rho = 0. 38; p<.01) and the hypothesis on level of pay and sickness presenteeism is also supported (rho = -0.22; p<0.01). CONCLUSIONS: Members of occupational groups whose everyday tasks are to provide care or welfare services, or teach or instruct, have a substantially increased risk of being at work when sick. The link between difficulties in replacement or finding a stand in and sickness presenteeism is confirmed by study results. The categories with high sickness presenteeism experience symptoms more often than those without presenteeism. The most common combination is low monthly income, high sickness absenteeism and high sickness presenteeism. PMID- 10846193 TI - Impact of socioeconomic status on coronary mortality in people with symptoms, electrocardiographic abnormalities, both or neither: the original Whitehall study 25 year follow up. AB - OBJECTIVES: To determine the impact of socioeconomic status (SES) on coronary heart disease (CHD) mortality in people with and without prevalent CHD at baseline. DESIGN: Cohort study with 25 year follow up; prevalent CHD was defined by Q, ST or T wave electrocardiographic (ECG) abnormalities or symptoms (defined by the Rose chest pain questionnaire and self reported doctor diagnosis) or both. SES was defined by four civil service employment grades. SETTING: London. PARTICIPANTS: 17 907 male civil servants aged 40-69 years. MAIN OUTCOME MEASURES: CHD mortality (n=2695 deaths). RESULTS: The lowest versus highest employment grade was associated with increased CHD mortality (age adjusted hazard ratio 1.56 (95% CI 1.2, 2.1)), prevalence of symptoms and, among symptomatic participants only, the prevalence of Q, ST or T abnormalities. Thirty one per cent of CHD deaths occurred in participants with prevalent CHD at baseline. Among participants without Q, ST or T abnormality employment grade was associated with CHD mortality; the hazard ratios (lowest v highest grade) adjusted for age, systolic and diastolic blood pressure were 1.72 (95% CI 1.4, 2.1) for asymptomatic and 1.52 (95% CI 1.1, 2.1) for symptomatic participants; among participants with Q, ST or T abnormality the corresponding hazard ratios were 1.46 (95% CI 0.7, 2.9) and 1.14 (95% CI 0.6, 2.0) respectively. CONCLUSIONS: SES was inversely associated with CHD mortality in civil servants with and without prevalent CHD at baseline. Further distinguishing the relative contribution of SES to the initiation and progression of CHD requires repeated measures studies of pre-clinical and clinical measures of CHD. PMID- 10846194 TI - Socioeconomic status and injury mortality: individual and neighbourhood determinants. AB - STUDY OBJECTIVE: This study examined both individual and neighbourhood correlates of injury mortality to better understand the contribution of socioeconomic status to cause specific injury mortality. Of particular interest was whether neighbourhood effects remained after adjusting for individual demographic characteristics and socioeconomic status. DESIGN: Census tract data (measuring small area socioeconomic status, racial concentration, residential stability, urbanisation, and family structure) was merged with the National Health Interview Survey (NHIS) and a file that links the respondents to subsequent follow up of vital status and cause of death data. Cox proportional hazards models were specified to determine individual and neighbourhood effects on homicide, suicide, motor vehicle deaths, and other external causes. Variances are adjusted for the clustered sample design of the NHIS. SETTING: United States, 1987-1994, with follow up to the end of 1995. PARTICIPANTS: From a sample of 472 364 persons ages 18-64, there were 1195 injury related deaths over the follow up period. MAIN RESULTS: Individual level effects were generally robust to the inclusion of neighbourhood level variables in the models. Neighbourhood characteristics had independent effects on the outcome even after adjustment for individual variability. For example, there was approximately a twofold increased risk of homicide associated with living in a neighborhood characterised by low socioeconomic status, after adjusting for individual demographic and socioeconomic characteristics. CONCLUSIONS: Social inequalities in injury mortality exist for both persons and places. Policies or interventions aimed at preventing or controlling injuries should take into account not only the socioeconomic characteristics of people but also of the places in which they live. PMID- 10846195 TI - Mortality differences by parental social class from childhood to adulthood. AB - STUDY OBJECTIVE: To examine mortality differences by parental social class and cause of death from age 5 to age 34. DESIGN: Register-based follow up study based on census records for 1985 and 1990 linked with death records for the period 1987 95. SETTING AND SUBJECTS: The study covers all males and females in non-manual and manual classes in Finland aged 5-34 years in 1987-95 (8135 deaths). Parental social class is defined on the basis of the occupation of the head of household at the time the child was 0-14 years. MAIN OUTCOME MEASURES: All cause mortality, mortality from diseases, mortality from accidents and violence, and alcohol related mortality during the period 1987-95. MAIN RESULTS: At ages 5-14 there is no systematic gradient in mortality by parental social class. Both absolute and relative differences increase with age. The relative rate of male all cause mortality among manual class descendants at ages 25-29 compared with that of upper non-manual class descendants is 1.60 (95% CI 1.37, 1.86). At ages 30-34 the relative rate among males is 1.95 (95 % CI 1.58, 2.42) and among females 1.47 (95% CI 1. 03, 2.10). Among males alcohol related causes of death account for 70% of the excess mortality of sons of manual class parents compared with sons of upper non-manual class parents at ages 25-34. At ages 25-34, both among females and males, the contribution of diseases to the mortality difference increases. CONCLUSIONS: Parental social class has an impact on mortality after childhood mainly through health related behaviours and lifestyles up to age 34. PMID- 10846196 TI - Health conditions and residential concentration of poverty: a study in Rio de Janeiro, Brazil. AB - STUDY OBJECTIVES: To establish the geographical relation of health conditions to socioeconomic status in the city of Rio de Janeiro, Brazil. DESIGN: All reported deaths in the municipality of Rio de Janeiro, from 1987 to 1995, obtained from the Mortality Information System, were considered in the study. The 24 "administrative regions" that compose the city were used as the geographical units. A geographical information system (GIS) was used to link mortality data and population census data, and allowed the authors to establish the geographical pattern of the health indicators considered in this study: "infant mortality rate"; "standardised mortality rate"; "life expectancy" and "homicide rate". Information on location of low income communities (slums) was also provided by the GIS. A varimax rotation principal component analysis combined information on socioeconomic conditions and provided a two dimension basis to assess contextual variation. MAIN RESULTS: The 24 administrative regions were aggregated into three different clusters, identified as relevant to reflect the socioeconomic variation. Almost all health indicator thematic maps showed the same socioeconomic stratification pattern. The worst health situation was found in the cluster composed of the harbour area and northern vicinity, precisely in the sector where the highest concentration of slum residents are present. This sector of the city exhibited an extremely high homicide rate and a seven year lower life expectancy than the remainder of the city. The sector that concentrates affluence, composed of the geographical units located along the coast, showed the best health situation. Intermediate health conditions were found in the west area, which also has poor living standards but low concentration of slums. CONCLUSIONS: The findings suggest that social and organisation characteristics of low income communities may have a relevant role in understanding health variations. Local health and other social programmes specifically targeting these communities are recommended. PMID- 10846197 TI - Exploring health preferences in sociodemographic and health related groups through the paired comparison of the items of the Nottingham health profile. AB - BACKGROUND: Preference weighted measures of health related quality of life are necessary for cost effectiveness calculations involving quality of life adjustment. There are conflicting data about the influence of factors such as sociodemographic and health related variables on health preferences. STUDY OBJECTIVE: The relative values attached to the items of the Spanish version of the Nottingham Health Profile (NHP) were assessed to make comparisons across social and health subgroups. DESIGN AND PARTICIPANTS: Preference values were obtained in sets of 250 to 253 persons (total n=1258) using the method of paired comparisons after all possible pairs of NHP items had been presented to respondents for judgement of severity. chi(2) Tests and Spearman's correlations among item ranks were calculated. MAIN RESULTS: Findings show that preferences elicited with the method of paired comparisons are consistent and independent of the sample from which they are obtained (mean correlation coefficients across subgroups range from 0.87 to 0.96). Conclusion-The evaluation of health did not seem to be related to sociodemographic variables (gender, age, social class) or to the health status of the respondents, suggesting that health preferences are stable across different populations. PMID- 10846198 TI - Validity of self reported utilisation of primary health care services in an urban population in Spain. AB - STUDY OBJECTIVE: To assess the validity and factors related with the validity of self reported numbers of visits to a primary health care centre, in comparison with the recorded number. DESIGN: Cross sectional study. SETTING: The urban area served by the Zaidin-Sur Primary Health Care Centre (Granada, Spain). PARTICIPANTS: Two population samples (236 high users and 420 normal users) who were seen at the centre from 1985 to 1991 were interviewed in 1993. MAIN RESULTS: A net tendency to overreport the actual number of visits was observed. Absolute concordance between self reported and recorded utilisation decreased as time interval lengthened, although this mainly reflected the increase in maximum variability both with time interval length and with the number of recorded visits. Corrected Spearman rho coefficients obtained between the number of self reported and recorded visits ranged from 0.602 for the two weeks before the interview to 0.678 for the year before. Regression slopes of self reported utilisation upon recorded utilisation did not change between periods. In multiple regression analyses the actual number of visits was the main factor associated with both underreporting and overreporting. Older age was also significantly associated with underreporting. Poor health status and high satisfaction with health care were significantly associated with overreporting. CONCLUSIONS: There was a substantial degree of inaccuracy in self reported utilisation, with a net tendency to overreport the number of visits. In relative terms, however, accuracy of self reports did not seem to decrease appreciably as the recall time lengthened. To compare the accuracy of different measures, it is important to take into account the maximum variability of each one. Otherwise, contradictory results may be obtained. PMID- 10846199 TI - Emergence and preservation of a chronically sick building. AB - STUDY OBJECTIVE: To investigate the merits of case studies as complementary methodological approaches in the study of the sick building syndrome. SETTING AND PARTICIPANTS: A Swedish office building with longstanding health problems, and its inhabitants. DESIGN: This paper is a case study based both on historical and present, quantitative as well as qualitative, documentary material, produced over the years by distinct parties, and on semi-structured interviews. RESULTS: Long drawn conflictive processes within the building were identified. It was revealed that the organisation for dealing with environmental problems was split, and ineffective with poor patterns of communication. It was suggested that this generated a situation of chronic stress leading to the persistence of symptoms. CONCLUSIONS: By their capacity to identify internal processes within building contexts, case study methodology can contribute to a better understanding and management of sick building syndrome. The results of this study suggest that psychosocial factors, among them organisational structures and communication patterns, should be given close attention. PMID- 10846200 TI - Soft tissue injury resulting from falling predicts a future major falling injury in the home dwelling elderly. PMID- 10846201 TI - DNA uptake sequences in Helicobacter pylori. PMID- 10846202 TI - Authors' reply PMID- 10846203 TI - PCR-based detection of mobile genetic elements in total community DNA. PMID- 10846204 TI - Teichoic acids in bacterial coaggregation. PMID- 10846205 TI - Cyanide inhibits respiration yet stimulates aerobic growth of Zymomonas mobilis. AB - Potassium cyanide at submillimolar concentrations (20-500 microM) inhibited the high respiration rates of aerobic cultures of Zymomonas mobilis but, remarkably, stimulated culture growth. In batch culture, after an extended lag phase, exponential growth persisted longer, resulting in higher biomass densities. In aerobic chemostat cultures, elevated biomass concentration was observed in the presence of cyanide. This growth stimulation effect is attributed to decreased production of the inhibitory metabolite acetaldehyde at lowered respiration rates, when more reducing equivalents are channelled to alcohol dehydrogenase. Growth in the presence of cyanide did not alter the membrane cytochrome content. In non-growing cyanide-preincubated cells, with ethanol as the respiratory substrate, cyanide increased ATP levels; in such cells, a large part of the cyanide-sensitive respiration was inhibited within a few seconds after ethanol addition, while inhibition of the rest of respiration took several minutes. The more cyanide-sensitive respiration was apparently energy-nongenerating, and was absent in membrane preparations. Pelleting of membranes from cell-free extracts produced 'soluble' fractions in which a b-type haem was detectable by reduced minus oxidized difference spectroscopy. The function of the Z. mobilis respiratory chain in cell growth and respiratory protection, and the possible physiological role of aerobic generation of inhibitory metabolites, are discussed. PMID- 10846206 TI - Quantitative detection of Streptococcus pneumoniae cells harbouring single or multiple copies of the gene encoding the green fluorescent protein. AB - A modified gfp gene from Aequorea victoria, encoding a variant of the green fluorescent protein (GFP), was subcloned into the mobilizable plasmid pMV158. gfp was placed under the control of the inducible P(M) promoter of the Streptococcus pneumoniae gene malM, cloned in plasmid pLS70. The P(M) promoter is regulated by the product of the pneumococcal malR gene, which is inactivated by growing the cells in maltose-containing media. By homologous recombination, the P(M)-gfp construction was integrated into the host chromosome in a single copy. In both conditions (single and multiple copies), the pneumococcal cells were able to express GFP in an inducible or constitutive form, depending on whether the S. pneumoniae strain harboured a wild-type or a mutant malR gene. Quantification of the levels of GFP expressed by cultures supplemented with sucrose or maltose as carbon sources was feasible by fluorescence spectroscopy. Phase-contrast and fluorescence microscopy allowed pneumococcal cells expressing GFP in mixed cultures to be distinguished from those not carrying the gfp gene. PMID- 10846207 TI - Comparison of conserved structural and regulatory domains within divergent 16S rRNA-23S rRNA spacer sequences of cyanobacteria. AB - PCR amplification of the internal transcribed spacer (ITS) between the 16S rRNA and 23S rRNA genes of the cyanobacterium NOSTOC: PCC 7120 gave three products. Two represented true ITS regions of different sizes, while the third was a heteroduplex. The longer spacer (ITS-L) contained 512 nucleotides and carried tRNA(Ile) and tRNA(Ala) genes, separated by a large stem-loop structure (V2) composed of short tandemly repeated repetitive sequences. Both tRNA genes, and the 5' half of the intervening stem, were absent from the shorter spacer (ITS-S), of length 283 nucleotides, which was otherwise almost completely identical to ITS L. The two spacer regions of NOSTOC: PCC 7120 were aligned to published ITS sequences of cyanobacteria, the cyanelle of Cyanophora paradoxa and Escherichia coli. Although the ITS regions of cyanobacteria vary in length from 283 to 545 nucleotides and contain either both tRNA(Ile) and tRNA(Ala) genes, only the tRNA(Ile) gene, or neither, there is no correlation between ITS size and coding capacity for tRNAs. Putative secondary structures were determined for the deduced transcripts of the rrn operons of several cyanobacteria and were compared to that of E. coli. Highly conserved motifs important for folding and for maturation of the rRNA transcripts were identified, and regions homologous to bacterial antiterminators (box B-box A) were located. The conserved and variable regions of the cyanobacterial ITS are potential targets of PCR primers and oligonucleotide probes for detection and identification of cyanobacteria at different taxonomic levels. PMID- 10846208 TI - Relative abundance of Archaea and Bacteria along a thermal gradient of a shallow water hydrothermal vent quantified by rRNA slot-blot hybridization. AB - Slot-blot hybridization of rRNA with domain-specific oligonucleotide probes targeting the 16S rRNA of Archaea and Bacteria was utilized to assess the relative abundance of these domains along a thermal gradient at a shallow submarine hydrothermal vent near Milos Island (Greece). The highest prokaryotic rRNA concentrations (defined as the sum of bacterial and archaeal rRNA) were found in the uppermost sediment surface (0-20 mm), decreasing strongly with depth. This indicates that the microbial activity was mainly occurring in the surface layer of this hydrothermal vent. Furthermore, rRNA concentrations were higher in regions closer to the vent, suggesting that the hydrothermal activity stimulated microbial activity. Archaea seemed to be a minor component of the microbial community at this vent site, even in the zones with higher temperatures. Bacteria made up at least 78% (mean 95%) of the prokaryotic rRNA. However, along the steepest temperature gradient, the proportion of archaeal rRNA increased. Nevertheless, even in the hottest sediment layer where a quantification was possible (in situ temperature 82 degrees C) archaeal rRNA made up only 11.9% of the prokaryotic rRNA. This suggests that Archaea were generally of minor importance at this vent site and were probably restricted to a narrow niche. The factors that allow Bacteria to dominate in a high temperature environment that was once believed to be the realm of Archaea remain elusive. PMID- 10846209 TI - Phylogenetic and physiological diversity of Arthrobacter strains isolated from unconsolidated subsurface sediments. AB - Forty strains of Gram-positive, aerobic, heterotrophic bacteria isolated from saturated subsurface lacustrine, paleosol and fluvial sediments at the US Department of Energy's Hanford Site in south central Washington State were characterized by phylogenetic analysis of 16S rRNA gene sequences and by determination of selected morphological, physiological and biochemical traits. Phylogenetic analyses of 16S rDNA sequences from subsurface isolates in the context of similar sequences from previously described bacterial species indicated that 38 of the subsurface strains were most closely related to Arthrobacter: The other two strains appeared to be most closely related to Kocuria. The subsurface isolates fell into seven phylogenetically coherent and distinct clusters, indicating that there was a significant degree of diversity among them. Additional diversity was detected by analysis of cellular fatty acids and physiological traits. The general morphological, physiological and biochemical traits of the subsurface strains were consistent with those of Arthrobacter, Micrococcus and genera recently separated from Micrococcus, such as Kocuria. Some of the subsurface strains were phylogenetically closely related to certain species of Arthrobacter. (16S rDNA sequence similarities >99%). However, most of the subsurface isolates did not cluster with previously established species in phylogenetic analyses of 16S rRNA gene sequences or with hierarchical cluster analysis of cellular fatty acid profiles. Moreover, many of the subsurface isolates that were most closely related to Arthrobacter. also differed from all established species of that genus in several of their specific physiological characteristics. Most of the subsurface isolates, then, are likely to be novel strains or species of Arthrobacter. PMID- 10846210 TI - Gliding mutants of Mycoplasma mobile: relationships between motility and cell morphology, cell adhesion and microcolony formation. AB - The present study characterizes gliding motility mutants of Mycoplasma mobile which were obtained by UV irradiation. They were identified by their abnormal colony shapes in 0.1% agar medium, showing a reduced number of satellite colonies compared to the wild-type. A total of ten mutants were isolated based on their colony phenotype. Using dark-field and electron microscopy, two classes of mutants, group I and group II, were defined. Cells of group I mutants had irregular, flexible and sometimes elongated head-like structures and showed a tendency to aggregate. Neither binding to glass nor gliding motility was observed in these mutants. Cells of group II mutants were rather spherical in shape, with the long axis reduced to 80% and the short axis enlarged to 120% of that of wild type cells, respectively. Their gliding speed was 20% faster than that of wild type cells. Three of the ten mutants remained unclassified. Mutant m6 had a reduced binding activity to glass and a reduced gliding motility with 50% of the speed of the wild-type strain. The ability of wild-type and mutant colonies to adsorb erythrocytes was found to correlate with the binding activity required for gliding, indicating that mycoplasma gliding depends on cytadherence-associated components. Finally, the ability to form microcolonies on surfaces was shown to correlate with the gliding activity, suggesting a certain role of gliding motility in the parasitic life-cycle of mycoplasmas. PMID- 10846211 TI - Identification of a novel gene, fimV, involved in twitching motility in Pseudomonas aeruginosa. AB - Transposon mutagenesis was used to identify a new locus required for twitching motility in Pseudomonas aeruginosa. Four Tn5-B21 mutants which lacked twitching motility and a fifth which exhibited impaired motility were found to map to the same KPN:I restriction fragment at approximately 40 min on the P. aeruginosa genome. Cloning and sequencing studies showed that all five transposon insertions occurred within the same 2.8 kb ORF, which was termed fimV. The product of this gene has a putative peptidoglycan-binding domain, predicted transmembrane domains, a highly acidic C terminus and anomalous electrophoretic migration, indicating unusual primary or secondary structure. The P. aeruginosa genome also possesses a paralogue of fimV. Homologues of fimV were also found in the sequenced genomes of the other type-IV-fimbriated bacteria Neisseria gonorrhoeae, Neisseria meningitidis, Legionella pneumophila and Vibrio cholerae, but not in those of other bacteria which lack type IV fimbriae. A fimV homologue was also found in the genome sequence of Shewanella putrefaciens, along with many other homologues of type IV fimbrial genes, indicating that this bacterium is also likely to produce type IV fimbriae. Wild-type twitching motility was restored to fimV mutants by complementation in a dosage-dependent manner. Overexpression of fimV resulted in an unusual phenotype where the cells were massively elongated and migrated in large convoys at the periphery of the colony. It is suggested that FimV may be involved in remodelling of the peptidoglycan layer to enable assembly of the type IV fimbrial structure and machinery. PMID- 10846212 TI - Intimin from enteropathogenic Escherichia coli mediates remodelling of the eukaryotic cell surface. AB - Adhesion to cultured epithelial cells by enteropathogenic Escherichia coli (EPEC) is associated with extensive rearrangement of the host cell cytoskeleton. Evidence has been presented that EPEC adhesion is associated with activation of signal transduction pathways leading to production of a characteristic histopathological feature known as the attaching and effacing (A/E) lesion. A/E lesion formation requires intimin, an EPEC adhesion molecule and several EPEC secreted proteins (EspA, B, D and Tir) involved in cell signalling and protein translocation. In this study it is shown that HEp-2 cells respond during the early stages of infection with two wild-type EPEC strains (B171 and E2348/69) by producing microvillus-like processes (MLP) at the site of initial bacterial adherence. Intimin appears to play a key role in MLP elongation. At later stages of infection with these wild-type EPEC strains, when A/E lesions have formed, the MLP were reduced in number and length to appear as at time zero, and the cell surface in the vicinity of bacterial clusters appeared unaffected. In contrast, infection with EspA- or EspB-negative, but intimin-positive, EPEC strains (UMD872 and UMD864, respectively) resulted in enhanced MLP proliferation and formation of 'cage-like' structures engulfing the bacteria. Inoculating HEp-2 cells with intimin-coated latex spheres induced similar 'cage-like' structures. Caco-2 cells did not show intimin-induced microvillus elongation in response to EPEC infection, although microvillus effacement and reduction in number occurred. Similar phenomena appeared on B171 and E2348/69 infection of paediatric intestine using in vitro organ culture, i.e. elongated microvilli were seen in association with small colonies and at the periphery of large localized colonies, along with evidence of microvillus breakdown and debris in the colony centre. These results show that intimin activates signal transduction pathways involved in the remodelling of the eukaryotic cell surface, probably via binding to a receptor encoded by the host cell. PMID- 10846213 TI - Identification of novel loci involved in entry by Legionella pneumophila. AB - Legionella pneumophila is primarily an intracellular pathogen during infection; thus, the mechanisms of entry into host cells are likely to be important for pathogenesis. Several L. pneumophila mutants that display an enhanced-entry (Enh) phenotype were isolated by selecting for bacteria that enter host cells at a higher frequency than wild-type. In the course of characterizing the genetic basis of one of these mutants, C3, a strategy was developed for the isolation of laboratory-media-repressed virulence determinants from L. pneumophila. Screens for dominant mutations using a genomic DNA library from C3 resulted in the isolation of three cosmids that confer an Enh phenotype to wild-type L. pneumophila. Transposon mutagenesis of these cosmids allowed identification of three loci that affect entry. Analysis of the putative proteins encoded by these loci, designated rtxA and enhC, demonstrated similarity to repeats in the structural toxin protein and the secreted Sel-1 protein from Caenorhabditis elegans, respectively. L. pneumophila rtxA and enhC mutants display significantly reduced entry into host cells, compared to wild-type bacteria. The phenotype that the cosmids containing these loci confer is most likely due to elevated expression resulting from their presence on multicopy vectors. The use of increased gene copy number to overexpress genes that are normally repressed under laboratory growth conditions is generally applicable to the isolation of virulence determinants from L. pneumophila and other bacterial pathogens. PMID- 10846214 TI - Streptococcus equi with truncated M-proteins isolated from outwardly healthy horses. AB - The M-protein genes of Streptococcus equi isolated from 17 outwardly healthy horses after 4 strangles outbreaks had ended, including a quarantined animal, were compared with those of S. equi isolates from 167 active cases of strangles across 4 countries. The healthy horses included 16 persistent S. equi carriers, at least one from each of the four outbreaks. These carriers, despite being outwardly healthy, had empyema of the guttural pouch(es), an enlargement of the equine Eustachian tube. A persistent carrier from two of these outbreaks, the quarantined animal and a healthy animal with normal guttural pouches, from which S. equi was isolated only once, were colonized by variant S. equi with truncated M-protein genes (24% of outwardly healthy animals with S. equi). The truncated M protein genes had in-frame deletions in slightly different positions between the signal sequence and the central repeat region, equivalent to approximately 20% of the mature expressed protein. Immunoblotting with antibody to recombinant M protein confirmed that the variants expressed a truncated form of the M-protein. In contrast to the outwardly healthy S. equi carriers, only 1/167 of S. equi isolates from strangles cases possessed a truncated M-protein gene (<1%; Fisher's exact test, P=0.0002). Compared with isolates from healthy horses with a truncated M-protein, much more of the N terminus of the truncated M-protein was retained in the variant S. equi from a strangles case. Variant S. equi from outwardly healthy animals were more susceptible to phagocytosis by neutrophils in vitro than typical isolates. This is the first report of detection of S. equi with a truncated M-protein. The distribution of the variants between strangles cases and carriers suggests that the 80% of the M-protein retained in the variants may contribute to colonization whilst the deleted portion of the gene may be needed for full virulence. PMID- 10846215 TI - Tripartite haemolysin BL: isolation and characterization of two distinct homologous sets of components from a single Bacillus cereus isolate. AB - Haemolysin BL (HBL), a three-component enterotoxic/necrotizing/vascular permeability toxin, is a likely virulence factor of Bacillus cereus diarrhoeal food poisoning and necrotic infections. This paper describes the isolation of two distinct homologous sets of all three HBL components from a single B. cereus isolate, MGBC 145. The proteins of one set (designated HBL, consisting of B, L(1) and L(2)), were about 87-100% identical in N-terminal amino acid sequences to their respective prototype components from strain F837/76, and the proteins of the homologous set (HBL(a), consisting of B(a), L(1a) and L(2a)) were all about 62-65% identical. Only the latter homologues differed immunochemically and physicochemically from the prototypes. HBL and HBL(a) exhibited similar haemolytic and vascular permeability potencies, and the homologues could be interchanged freely. There were no notable differences in activity between the L component homologues. However, components B and B(a) were significantly different. Both were secreted as monomers, but unlike B, B(a) was isolated as a relatively inactive complex that could be reactivated with urea. When B(a) was substituted for B in gel-diffusion assays the distinct discontinuous haemolysis pattern typical of the presence of B did not occur. In suspension assays, excess B inhibited the haemolysis of B-primed cells by L(1) (as previously described), but not that of B(a)-primed cells. Excess B(a) had the opposite effect and enhanced lysis of B(a)-primed cells, but not that of B-primed cells. These differences reveal details about how the toxin components interact on target cell membranes. The authors' observations indicate that HBL represents a new family of multicomponent toxins that was generated by a process of gene and operon duplication that occurred either intracellularly or by horizontal transfer, and raise the possibility of the existence of other related toxins in the genetically diverse B. cereus taxonomic group. PMID- 10846216 TI - Restriction fragment differential display of pediocin-resistant Listeria monocytogenes 412 mutants shows consistent overexpression of a putative beta glucoside-specific PTS system. AB - Pediocin PA-1, which is a bacteriocin produced by lactic acid bacteria, has potential as a biopreservative of food. However, such use may lead to the development of resistance in the target organism. Gene expression in two independent pediocin-resistant mutants of Listeria monocytogenes 412 was compared to the original isolate by restriction fragment differential display PCR (RFDD PCR). This method amplifies cDNA restriction fragments under stringent PCR conditions, enabled by the use of specific primers complementary to ligated adaptor sequences. RFDD-PCR was very well suited for analysis of listerial gene expression, giving reproducible PCR product profiles. Three gene fragments having increased expression in both resistant mutants were identified. All three had homology to components of beta-glucoside-specific phosphoenolpyruvate-dependent phosphotransferase systems (PTS), one fragment having homology to enzyme II permeases, and the two others to phospho-beta-glucosidases. Overexpression of the putative PTS system was consistently observed in 10 additional pediocin-resistant mutants, isolated at different pH, salt content and temperature. The results suggest that RFDD-PCR is a strong approach for the analysis of prokaryotic gene expression and that the putative beta-glucoside-specific PTS system is involved in mediating pediocin resistance. PMID- 10846217 TI - Three multidomain esterases from the cellulolytic rumen anaerobe Ruminococcus flavefaciens 17 that carry divergent dockerin sequences. AB - Three enzymes carrying esterase domains have been identified in the rumen cellulolytic anaerobe Ruminococcus flavefaciens 17. The newly characterized CesA gene product (768 amino acids) includes an N-terminal acetylesterase domain and an unidentified C-terminal domain, while the previously characterized XynB enzyme (781 amino acids) includes an internal acetylesterase domain in addition to its N terminal xylanase catalytic domain. A third gene, xynE, is predicted to encode a multidomain enzyme of 792 amino acids including a family 11 xylanase domain and a C-terminal esterase domain. The esterase domains from CesA and XynB share significant sequence identity (44%) and belong to carbohydrate esterase family 3; both domains are shown here to be capable of deacetylating acetylated xylans, but no evidence was found for ferulic acid esterase activity. The esterase domain of XynE, however, shares 42% amino acid identity with a family 1 phenolic acid esterase domain identified from Clostridum thermocellum XynZ. XynB, XynE and CesA all contain dockerin-like regions in addition to their catalytic domains, suggesting that these enzymes form part of a cellulosome-like multienzyme complex. The dockerin sequences of CesA and XynE differ significantly from those previously described in R. flavefaciens polysaccharidases, including XynB, suggesting that they might represent distinct dockerin specificities. PMID- 10846218 TI - Structure-function analysis of NADPH:nitrate reductase from Aspergillus nidulans: analysis of altered pyridine nucleotide specificity in vivo. AB - Nitrate reductase (NaR) catalyses the reduction of nitrate to nitrite via a two electron transfer. In fungi, the electron donor for NaR is NADPH whereas plants can have two enzymes, NADH:NaR and a bispecific NAD(P)H:NaR. PCR mutagenesis was employed to introduce mutations into the niaD gene of Aspergillus nidulans in order to identify residues involved in co-enzyme specificity. The niaD3000 mutation (NiaD T813D, K814Q) altered co-enzyme specificity: the new enzyme had high levels of NADH:NaR activity in vitro, whilst all NADPH-associated activity was lost. However, strains carrying this mutation did not grow on nitrate. Enzyme assays suggested that this was not due to inhibition of the mutant enzyme by NADPH. All revertants of the niaD3000 mutants had restored NADPH activity and lost NADH activity. Sequence analysis of these revertants showed that they all contained a single amino acid change at Asp-813, suggesting that this position is crucial to co-enzyme specificity. Further studies have shown that the mutant enzyme was not protected from deactivation by either co-factor in cell-free extracts (unlike the wild-type), and that induction of the glucose-6-phosphate dehydrogenase occurred independently of NADPH levels. These data highlight the importance of functional tests in vivo under physiological conditions. PMID- 10846219 TI - Expression of the nifA gene of Herbaspirillum seropedicae: role of the NtrC and NifA binding sites and of the -24/-12 promoter element. AB - The nifA promoter of Herbaspirillum seropedicae contains potential NtrC, NifA and IHF binding sites together with a -12/-24 sigma(N)-dependent promoter. This region has now been investigated by deletion mutagenesis for the effect of NtrC and NifA on the expression of a nifA::lacZ fusion. A 5' end to the RNA was identified at position 641, 12 bp downstream from the -12/-24 promoter. Footprinting experiments showed that the G residues at positions -26 and -9 are hypermethylated, and that the region from -10 to +10 is partially melted under nitrogen-fixing conditions, confirming that this is the active nifA promoter. In H. seropedicae nifA expression from the sigma(N)-dependent promoter is repressed by fixed nitrogen but not by oxygen and is probably activated by the NtrC protein. NifA protein is apparently not essential for nifA expression but it can still bind the NifA upstream activating sequence. PMID- 10846220 TI - Identification of a second lipase gene, gehD, in Staphylococcus epidermidis: comparison of sequence with those of other staphylococcal lipases. AB - The identification and molecular characterization of a previously unidentified lipase, gehD, from the human cutaneous commensal Staphylococcus epidermidis is reported. A lipase-GehC-deficient but otherwise isogenic mutant of S. epidermidis 9 was constructed by allele replacement. However, the mutant was found to retain 50% of the wild-type lipase activity in liquid culture. Rescreening of a genomic library revealed the presence of a second lipase gene, gehD, which was subsequently mapped and sequenced. In common with other staphylococcal lipases, GehD appeared to be translated as a 650-700 amino acid precursor which is processed post-translationally to an extracellular mature lipase of 360 amino acids with a size of approximately 45 kDa. Comparison of the amino acid sequence of GehD with those of other staphylococcal lipases revealed a high level of conservation between the mature lipase domains of different species. By hybridization studies, both gehC and gehD genes were found to be present in S. epidermidis isolates from both clinical and non-clinical backgrounds, but neither hybridized to DNA isolated from other staphylococcal strains. Construction of a phylogenetic tree and calculation of amino acid sequence homologies between mature lipases, however, suggested that the lipases of S. epidermidis may be more closely related to those of Staphylococcus aureus than to each other. PMID- 10846222 TI - The Fusobacterium nucleatum porin FomA possesses the general topology of the non specific porins. AB - FomA is a major non-specific porin of Fusobacterium nucleatum with no sequence similarity to other known porins. According to the topology model, the protein consists of 16 transmembrane beta-strands, connected by eight surface-exposed loops and seven periplasmic turns. In this study, the insertion mutagenesis approach was applied to probe the topology model. A Semliki Forest Virus (SFV) epitope was successfully inserted at 11 different sites of the FomA protein and a 6-aa insertion was successfully inserted at two different sites. Correct folding of the mutant proteins and proper incorporation into the outer membrane were assessed by heat modifiability and by an in vivo porin activity assay. Immunofluorescence microscopy analysis of intact cells, using mAbs directed against the inserted SFV epitope, revealed that three of the eight putative extracellular loops are indeed surface-exposed. Trypsin accessibility experiments confirmed the cell surface exposure of two additional loops. The results support the proposed topology model, showing that FomA possesses the general beta-barrel topology of the non-specific porins, with the interesting exception that the third loop does not seem to fulfil the role of a constriction loop. PMID- 10846221 TI - The Pseudomonas aeruginosa hscA gene encodes Hsc66, a DnaK homologue. AB - Under heat-stress conditions bacteria induce, among other heat-shock proteins, the Hsp70 molecular chaperone (DnaK), which is involved in protein stabilization. It has been shown in Escherichia coli that an Hsp70 homologue called Hsc66, which is widespread in bacteria, functions as a chaperone in vitro. This paper reports the isolation of a Pseudomonas aeruginosa W51D mutant (W51M22) by insertion of the mini-Tn5-Hg transposon, which was unable to grow on ethanol and other short chain alcohols as sole source of carbon. The transposon insertion in this mutant was shown to be located in the hscA gene encoding Hsc66. The inability of mutant W51M22 to use ethanol was complemented by the E. coli hscBA-fdx operon. The authors characterized the transcriptional arrangement of hscA, showing that it forms part of an operon with the upstream hscB gene, and that it is also expressed from its own promoter. These results are compatible with the P. aeruginosa Hsc66 protein being a functional molecular chaperone involved in the stabilization, in the presence of ethanol, of some proteins required for bacterial growth on short-chain alcohols. PMID- 10846223 TI - ctsR of Lactococcus lactis encodes a negative regulator of clp gene expression. AB - Bacteria undergo a complex programme of differential gene expression in response to stress. In Bacillus subtilis, it was recently shown that CtsR, a negative transcriptional regulator, mediates stress-induced expression of components of the Clp protease complex. In this study, a gene was identified in the Gram positive bacterium Lactococcus lactis that encodes a 17 kDa product with 38% identity to the CtsR protein of B. subtilis. By Northern analyses it was found that in a L. lactis strain carrying a large internal deletion of ctsR, including the region encoding a putative helix-turn-helix motif, the amounts of clpC, clpP, clpB and clpE mRNAs were increased 3-8-fold compared to those present in wild type L. lactis MG1363. In another ctsR mutant strain in which only one-third of CtsR was deleted, leaving the putative DNA-binding domain and the C-terminal 29 amino acids intact, only minor derepression of clp gene expression was observed and, furthermore, all the clp genes were still induced by heat. These results indicate that the amino acids of CtsR involved in temperature sensing are located either close to the DNA-binding domain or in the C-terminal part of the protein. Thus, in L. lactis in addition to B. subtilis, CtsR is a key regulator of heat shock-induced gene expression, suggesting that the presence of CtsR-homologous DNA-binding sites observed in many Gram-positive bacteria reflects functional heat-shock regulatory systems. PMID- 10846224 TI - The dnrO gene encodes a DNA-binding protein that regulates daunorubicin production in Streptomyces peucetius by controlling expression of the dnrN pseudo response regulator gene. AB - The dnrO gene is located adjacent to and divergently transcribed from the response regulator gene, dnrN, that activates the transcription of the dnrI gene, which in turn activates transcription of the daunorubicin biosynthesis genes in Streptomyces peucetius. Gene disruption and replacement of dnrO produced the dnrO::aphII mutant strain and resulted in the complete loss of daunorubicin biosynthesis. Suppression of the dnrO::aphII mutation by the introduction of dnrN or dnrI on a plasmid suggested that DnrO is required for the transcription of dnrN, whose product is known to be required for dnrI expression. These conclusions were supported by the effects of the dnrO mutation on expression of dnrO, dnrN and dnrI, as viewed by melC fusions to each of these regulatory genes. DnrO was overexpressed in Escherichia coli and the cell-free extract was used to conduct mobility shift DNA-binding assays. The results showed that DnrO binds specifically to the overlapping dnrN/dnrO(p1) promoter region. Thus, DnrO may regulate the expression of both the dnrN and dnrO genes. PMID- 10846225 TI - Genetic linkage of the vanB2 gene cluster to Tn5382 in vancomycin-resistant enterococci and characterization of two novel insertion sequences. AB - VanB-type vancomycin resistance is encoded by the vanB gene cluster, which disseminates by horizontal gene transfer and clonal spread of vancomycin resistant enterococci (VRE). Genetic linkage of the vanB gene cluster to transposon Tn5382 and the insertion sequences IS16 and IS256-like has previously been shown. In this study linkage of defined vanB gene cluster subtypes to these elements was examined. All the vanB2 subtype strains studied (n=14) revealed co hybridization of vanB and Tn5382, whereas the strains of vanB1 (n=8) and vanB3 (n=1) subtypes were Tn5382 negative. Conjugative cotransfer of the vanB2 gene cluster and Tn5382 was demonstrated for two strains. DNA sequencing of the vanX(B)-ORFC region in vanB2 strains confirmed that the vanB2 gene cluster is an integral part of Tn5382. No general pattern of linkage was observed with regard to IS16 and IS256-like. Two novel insertion sequences were identified in specific vanB2 subtype strains. (i) A 1611 bp element (ISEnfa110) was detected in the left flank of Tn5382. Its insertion site, lack of terminal inverted and direct repeats, and two conserved motifs in its putative transposase all conform to the conventions of the IS110 family. (ii) A 787 bp element (ISEnfa200) was detected in the vanS(B)-vanY(B) intergenic region. Its ORF encoded a putative protein with 60-70% identity to transposases of the IS200 family. No further copies of ISEnfa110 were found by colony hybridization of 181 enterococcal isolates, whereas ISEnfa200 was found in four additional vanB2 strains from the USA. The five strains had identical ISEnfa200 element insertion sites, and Tn5382 was located downstream from a pbp5 gene conferring high-level ampicillin resistance. These isolates showed related PFGE patterns, suggesting possible clonal spread of a VRE strain harbouring a Tn5382-vanB2-ISEnfa200 element linked to a pbp5 gene conferring ampicillin resistance. PMID- 10846226 TI - Characterization of transposon Tn1549, conferring VanB-type resistance in Enterococcus spp. AB - Transfer of VanB-type resistance to glycopeptides among enterococci has been reported to be associated with the movement of large chromosomal genetic elements or of plasmids. The authors report the characterization of the 34 kb transposon Tn1549 borne by a plasmid related to pAD1 and conferring vancomycin resistance in clinical isolates of Enterococcus spp. Tn1549 contained 30 ORFs and appeared to be organized like the Tn916 family of conjugative transposons into three functional regions: (i) the right end, implicated in the excision-integration process; (ii) the central part, in which the vanB2 operon replaces the tet(M) gene; and (iii) the left extremity, in which eight of the 18 ORFs could be implicated in the conjugative transfer. PMID- 10846227 TI - Moods PMID- 10846228 TI - [Quality control....more and always]. PMID- 10846229 TI - [Medical biology needs a boost]. PMID- 10846230 TI - [The reality at stake in the point of care approach: methodology of the future, a momentary fashionable procedure or just a basic swindle?]. PMID- 10846231 TI - [The wonderful history of neonatal screening]. AB - The concept of neonatal screening for metabolic disorders was launched by Robert Guthrie in 1963. A filter paper blood sample and a cheap and rapid analytical technique were factors that paved the road to success. Mass screening for phenylketonuria soon started to spread. Various additional screening procedures for other metabolic or endocrine conditions were subsequently developed. Most of developed countries have set up systematic neonatal screening programs and million of babies are screened each year. This review presents the main milestones of the fantastic history of neonatal screening but also the questions risen by many temptations to extend the screening to other disorders which not necessarily fulfil classic criteria for systematic search. Ethical and health economical aspects will be the most important issues for the development of neonatal screening in the next millennium. PMID- 10846232 TI - [Hereditary diseases of erythrocyte membrane: from clinical aspects to underlying genetical and molecular mechanisms]. AB - During the past years many studies on erythrocyte membrane structure and function have allowed for exponential development of this field. Inborn errors of red cell membrane include a variety of disorders characterized by alterations of the membrane skeleton or other membrane proteins. Most recently, many studies have been published using methods from molecular biology allowing for clarification of genotype phenotype relationship. This review outlines in details clinical aspects and organisational consequences of defects molecules of membrane proteins and skeleton and their contributions to the pathophysiology of hemolytic anaemia. PMID- 10846233 TI - [Drug resistance in Mycobacterium tuberculosis: diagnostic methods]. AB - The increasing frequency of multidrug-resistant strains of M. tuberculosis becomes dramatic in industrialized countries as well as in developing countries, particularly among patients infected with human immunodeficiency virus. It needs to formulate rapid strategies for diagnosing multidrug-resistant tuberculosis. For these new drug resistance, novel detection methods are developed in order to identify the resistant strains and to undertake efficacious antituberculosis therapies more rapidly. The phenotypic methods are based on the measurement of the microbial growth on nutritional supplement with antimicrobial agents; however, these proportional methods, such as the method in solid medium, the Bactec radiometric method or the MGIT method (mycobacterial growth indicator tube), are time consuming and give results in 5 to 21 days. In contrast, the genotypic tests, using knowledge of the genes involved in the resistance, reduce the time to detection of resistance from weeks to days. After amplification of the segment of the gene encoding the drug target by PCR, these methods are based on the identification of the different mutations conferring the antimicrobial resistance in M. tuberculosis. These methods are applied with success for detection of rifampicin resistance, conferring by mutations in a defined region of the rpoB gene for 99% of cases; on the contrary, results are less for other antituberculous drugs because of the insufficiency of knowledge of the molecular basis of drug resistance. PMID- 10846234 TI - [Carbohydrate deficient transferrin: what's new 20 years later?]. AB - The clinical diagnosis of alcohol abuse has frequently to be supported by biological data. These data help the practitioner to establish the diagnosis and to monitor the alcohol withdrawal and possible relapses. They also can help to demonstrate to the patient the existence of a denied alcohol abuse. CDT determination has been proposed for more than twenty years, but commercial kits are available since only 1990. A consensus has to be established concerning a reference method and the definition of the transferrin isoforms that should be included in the CDT. CDT is nevertheless considered in many studies as the best marker of alcohol abuse now available. Its sensitivity is at least equal to that of gamma glutamyltransferase (GGT), and its specificity remains very high even in the presence of other pathologies connected or not to the alcohol abuse. CDT is, despite the lack of standardisation and the cost of its determination, a basic marker of alcohol abuse more particularly in the circumstances of lack of efficiency of the other markers. PMID- 10846235 TI - [Diagnosis of malaria in non-endemic countries : value, limitations and complementarity of existing methods]. AB - The biological diagnosis of malaria is urgent to avoid rapid and fatal outcome. Every year in France, 5,000 imported malaria cases are observed. Thin stained blood smear microscopical examination remains the reference method of diagnosis; however its performance is linked to the professional competence of the biologists. Thus easier methods have been developed (QBC test). Some of them, limited to the diagnosis of malaria due to Plasmodium falciparum do not require highly skilled personal to perform or interpret (antigen detection on dipsticks, tests Parasight or cardboard, ICT Malaria Pf), but limitations and errors occurred. These different tests must be complementary methods of traditional diagnosis. In association with microscopical examinations, they provide rapid and efficient diagnosis of malaria in non-endemic areas. Relying on our experience, the best association is: QBC + thin blood smear and depending of results antigen detection (ParaSight F, ICT Malaria Pf). PMID- 10846236 TI - [Heparin-induced thrombocytopenia]. AB - Heparin-induced thrombocytopenia is a threatening complication of heparin treatment. The physio-pathological mechanism is the production of antibodies, the most frequent target of which is the complex heparin-platelet factor 4. These antibodies may activate the coagulation and lead to venous or arterial thromboembolic manifestations. Clinical features as well as functional and immunological tests are used for the diagnosis. The treatment consists in discontinuing heparin administration and in setting up an alternative treatment for which two drugs are indicated in France: Orgaran and Refludan. PMID- 10846237 TI - [Measurement of carotenoids by high pressure liquid chromatography: from difficulties to solutions]. AB - The measurement of serum carotenoids by HPLC has been largely improved during the last 10 years. However these techniques still require much time and skills, and direct application of published methods is rarely satisfying. We report here the difficulties that we met to transfer some HPLC methods described in the literature to our laboratories. We propose some solution to overcome the problems that we have encountered, our experience will perhaps help out other biologists. We reported also some results obtained in healthy populations. PMID- 10846238 TI - [Use of standard for quantitation of adhesion of polynuclear neutrophils by flow cytometry]. AB - Adherence receptors are essential for heterotypic (endothelial cell, platelet) polymorphonuclear neutrophil (PMN) interaction. Determination of their expression level give information about activation state and functionality of PMN. Use of flow cytometry associated with an immunolabeling standard, represented by beads coated by a determined amount of immunoglobulins (Qifikit, Dako), allows analysis of specific antibody binding capacity and gives information about antigen density. Using this methodology, the exploration of surface adherence receptors, L-selectin (CD62L) and b2-integrins (CD11a-c/CD18) from PMN unstimulated and incubated with pro-inflammatory stimuli, formyl-methionyl-leucyl-phenylalanine (fMLP) and tumor necrosis factor a (TNFa), allows, on the one hand, the establishment of basal expression values on resting PNN and on the other hand, the study of PNN reactivity. This method of quantification can be applied to clinical studies as adherence receptor deficiency syndromes or inflammatory, infectious and vascular diseases. PMID- 10846239 TI - [Epidemiology of neurodegenerative disorders : the example of dementia]. PMID- 10846240 TI - [Natural and molecular history of Alzheimer disease]. PMID- 10846241 TI - [Triplet expansion diseases]. PMID- 10846242 TI - [Cerebrospinal fluid in neurodegenerative dementia pathologies]. PMID- 10846243 TI - [Effect of hemolysis on determination and electrophoresis of serum proteins]. PMID- 10846244 TI - [Accuracy of antibiograms performed directly from urine incubation bottles of the Sebac system for diagnosing bacteriuria]. PMID- 10846245 TI - [Detection of oligoclonal IgG bands in in cerebrospinal fluid by immunofixation after electrophoretic migration in the automated Hydrasys sebia system]]. PMID- 10846246 TI - [Clinical enzymology in the 2000s: moving towards greater effectiveness for the doctor and the patient]. PMID- 10846247 TI - [Can biologists help to interpret biological findings?]. PMID- 10846248 TI - Photodermatology: progress, problems and prospects. AB - Photodermatology is a sub-specialty of photobiology. As such it includes all aspects of photobiology related to the skin ranging from sun exposure and its consequences (both short term and long term) to the therapeutic effects derived from exposure to natural or artificial radiation. In this review the terms photodermatology and photomedicine are used in a somewhat interchangeable fashion, although the former is really a portion of the latter. Four international journals are the primary sources for information - Photochemistry & Photobiology; Journal of Photochemistry & Photobiology (pt B); Photodermatology, Photomedicine & Photoimmunology; Journal of Investigative Dermatology. This review is structured so as to report on recent progress, existing problem areas and prospects for advances in the near future. PMID- 10846249 TI - Desmosomes and their autoimmune pathologies. AB - Desmosomes guarantee the integrity of the epidermis, by functioning both as an adhesive complex and as a cell-surface attachment site for the keratin intermediate filaments of the cytoskeleton. Considerable progress has been made in our knowledge of desmosomes and their components. The structure and function of many of the desmosomal molecules have been determined, and a number of the molecular interactions between desmosomal proteins have been elucidated. Desmosomal proteins are major antigens in pemphigus. Each type of pemphigus has its own antigenic targets, but in the last few years it has been shown that certain autoantibody populations are not restricted to just one form of pemphigus. The production of autoantibodies against multiple intracellular and extracellular desmosomal proteins, whose pathogenic role remains to be elucidated, suggests an overlapping distribution of antibody specificities among different forms of pemphigus. PMID- 10846250 TI - Surgical treatment of CHILD nevus. AB - We report a young girl with an unusual manifestation of CHILD syndrome in whom skin lesions showed involvement of the right side of her neck as well as symmetrically distributed ptychotropic involvement of the large body folds. Excision resulted in improvement and finally healing of skin lesions within the submammary folds, where breast reduction was also performed, whereas excision of axillary lesions and subsequent grafting with split skin turned out to be unsuccessful. PMID- 10846251 TI - Type 2 segmental manifestation of Hailey-Hailey disease: poor therapeutic response to dermabrasion is due to severe involvement of adnexal structures. AB - In autosomal dominant skin conditions, two different types of segmental manifestation can be distinguished. Type 1 represents heterozygosity for a postzygotic mutation, resulting in a degree of severity similar to that of the nonmosaic phenotype. Type 2 reflects loss of heterozygosity and shows an excessively pronounced involvement superimposed on the ordinary nonsegmental phenotype. We describe the clinical, histopathological and therapeutic aspects of the first case of type 2 segmental manifestation of Hailey-Hailey disease (HHD). A 24-year-old woman with a family history of HHD comprising four generations, presented with lesions of erythema and blistering arranged in a unilateral pattern following the lines of Blaschko. The disorder was first noted at the age of 3 months. At the age of 24 years, additional scattered symmetrical lesions involving the axillary and inguinal folds were noted. Histopathological examination of the severely involved linear skin areas revealed pronounced acantholysis within the deep adnexal structures, whereas clinically unaffected skin showed the typical histopathological features of the heterozygous phenotype with suprabasal clefting and acantholysis sparing the adnexae. Dermabrasion was performed in the areas of segmental involvement. During a follow-up period of one year, no recurrence was noted, but 18 months after dermabrasion a recurrence was present in the left submammary and left perianal regions. This therapeutic resistance to dermabrasion may be explained by the presence of acantholysis within the adnexal structures of the skin as found in type 2 segmental HHD. PMID- 10846252 TI - Efficacy and safety study of two zinc gluconate regimens in the treatment of inflammatory acne. AB - This double-blind study was conducted on 67 patients with inflammatory acne who received one of two zinc gluconate regimens (Rubozinc) for three months. One was a constant-dose regimen and the other included an initial three-week loading dose, but both regimens provided the same cumulative dose at three months. The primary assessment criteria was the change with respect to baseline in the total number of superficial inflammatory lesions (papules and pustules). The two treatment groups were not statistically significantly different, with respect to this criteria, after three, five, seven or thirteen weeks of treatment. Therefore, the regimen that included a loading-dose provided no additional benefit. The results of this study are in favor of the conventional therapeutic regimen of two capsules daily for three months, as defined in the marketing authorization. PMID- 10846253 TI - Serum adenosine deaminase levels in patients with psoriasis: a prospective case control study. AB - Adenosine deaminase (ADA) activity is a nonspecific marker of T cell activation. T cell activation is thought to play an important role in the pathogenesis of psoriasis. Our purpose was to assess the significance of serum ADA activity in psoriasis and its relevance to disease activity. ADA activity was determined with an enzymatic method in 25 patients with psoriasis and in 15 healthy subjects. These measurements were repeated for 10 patients after either PUVA or cyclosporin A treatments. Disease activity was estimated by the PASI scoring system. Serum ADA level was significantly elevated in patients with psoriasis compared to healthy subjects (p<0.05). There was a significant decrease in the ADA levels after treatment compared to pretreatment values in the same patients (p<0.05). There was no correlation between ADA levels and PASI scores. These results support the evidence that T cell activation is involved in the pathogenesis of psoriasis and that ADA may be valuable in the assessment of disease activity in psoriasis. PMID- 10846254 TI - Functional and morphological studies of photodamaged skin on the hands of middle aged Japanese golfers. AB - The skin of golfers' hands provides a suitable model to study the effect of chronic sun exposure, because one of their hands is exposed to the outer environment, especially sunlight, while the other one is always protected by a glove during play. Our purpose was to find out the influence of photodamage on the properties of the skin surface of middle-aged Japanese by using non-invasive methods. We measured hydration state, and water barrier function of the stratum corneum (SC) and the color of the skin of the dorsum of the hands. In a separate study, we evaluated the skin surface contour by using replicas taken from the skin in a slightly stretched or relaxed position. We found a significant decrease in hydration of the skin surface of the exposed skin as compared to that of the protected skin, whereas no such difference was found with transepidermal water loss, a parameter for water barrier function of the SC. Luminance of skin color was also reduced in the sun-exposed skin. Replica analysis revealed that large wrinkles developing in a relaxed position were more prominent on the exposed than on the protected skin, while fine furrows noted in a slightly stretched position were shallower on the former than the latter. The data obtained indicate that the chronically exposed skin of golfers' hands shows morphological and functional changes resulting from long time exposure to the outer environment especially sunlight. Furthermore, bioengineering non-invasive methods are found to be useful to detect early photodamage of the skin in a more quantitative fashion which is rather difficult to demonstrate clinically. PMID- 10846255 TI - The dermoscopic (7FFM) versus the clinical (ABCDE) diagnosis of small diameter melanoma. AB - The diagnosis of small diameter melanoma, that is early melanoma, is particularly difficult. For this reason we decided to evaluate the improvement given from our diagnostic dermoscopic method 7FFM to the clinical diagnosis, ABCDE rule, of small diameter melanoma. A retrospective study evaluating the clinical and the dermoscopic slides of 76 small diameter melanomas observed from January 1 1993 to December 31 1998, and of 524 small melanocytic nevi consecutively observed from September 1 1997 to September 30 1999, has been undertaken. The sensitivity and the specificity of the ABCDE rule and of our diagnostic dermoscopic method 7FFM in the diagnosis of small diameter melanoma have been calculated. The difference of diagnostic power between the two methods has been calculated with chi square test. The sensitivity and the specificity of the ABCDE rule in the diagnosis of small melanomas were 47.3% and 56%, while the sensitivity and the specificity of our method 7FFM were 68.8% and 86%. The difference of diagnostic power between the two methods was statistically significant: P<0.01 for both sensitivity and specificity. The sensitivity of the two methods together was 81.5% while the specificity of the two methods together was 50.6%. Our results show that our diagnostic dermoscopic method 7FFM improves both sensitivity and specificity in the clinical diagnosis of small diameter melanomas. Anyway the clinical and the dermoscopic diagnosis are not mutually exclusive as the best sensitivity is obtained with the two methods together. PMID- 10846256 TI - Drug related clinical pattern in fixed drug eruption. AB - Fixed drug eruption (FDE) represents a frequent type of drug eruption in Turkey. The aim of this open study is to analyze the clinical features with special emphasize on drug related pattern in our case series. Sixty-four cases with established FDE by oral provocation were clinically evaluated. Cotrimoxazole, a combination of sulfamethoxazole and trimethoprim, was the most common offender for FDE (75%), followed by naproxen sodium (12.5%), dipyrone (9.5%), dimenhydrinate (1.5%) and paracetamol (1.5%). Sensitivity to more than one drug was not observed. Cotrimoxazole-induced FDE was mainly located on male genitalia. Naproxen predominantly affected lips and face whereas dipyrone mainly caused FDE on trunk and extremities. Statistical analysis revealed a significant difference only for dipyrone versus cotrimoxazole over trunk and extremities (p = 0.03). Familial occurrence, symmetrical and asymmetrical nonpigmenting FDE, linear FDE, solitary plaque on the cheek, and "wandering" FDE were unusual findings of cotrimoxazole-induced FDE. Cotrimoxazole was the leading etiological agent in our series. Cotrimoxazole-induced FDE had some rarely or previously unreported features, but a significant relation between drugs and involved areas or clinical pattern could not be established. PMID- 10846257 TI - Cutaneous side effects induced by indinavir. AB - HIV-protease inhibitors demonstrated such high efficacy in short-term studies that they have been approved by the FDA, even though possible toxicity still needs further investigation. In the period between January 1997 and August 1998, 101 patients, staying at San Patrignano Medical Centre (Italy), received an HIV protease inhibitor (indinavir) plus two nucleoside reverse transcriptase inhibitors (NRTI's) selected from the following: AZT, didanosine, zalcitabine, lamivudine or stavudine. Seventy-three patients were male, 28 female and their ages ranged from 25 to 60 years, with an average of 34. At the end of the study, 84 patients were suitable for evaluation, as the other 17 dropped out for various reasons. Forty-eight patients (57.1%) developed cheilitis, 34 (40.5%) experienced diffuse cutaneous dryness and pruritus, 10 (11.9%) developed asteatotic dermatitis on the trunk, arms and thighs and another 10 (11.9%) complained of scalp defluvium. A severe alopecia was observed in only 1 patient (1.2%), while 6 reported that their body hair had become fairer, thinner and shed considerably. Multiple pyogenic granulomas were observed in the toenails of 5 patients (5. 9%). Softening of the nail plate was noted in 5 subjects as well. A peripheral lipodystrophy syndrome was noted in 12 patients (14.3%). Among these, one patient only developed a "buffalo hump" and another had diffused lipomatosis. The temporal relationship between the taking of indinavir and the onset of such cutaneous effects was striking. This was confirmed by the regression of symptoms in those patients who later discontinued indinavir. The emerging side effects of protease inhibitors require a multidisciplinary team for adequate diagnosis and treatment. Cutaneous toxicity involving the patient's own body image has a peculiar influence on compliance to the treatment and the patient's quality of life. PMID- 10846258 TI - The combined occurrence of macular amyloidosis and prurigo nodularis. AB - The association of prurigo nodularis (PN) and macular amyloidosis (MA) has not been reported before. Although pruritus related frictional trauma is a well-known cause of PN, its role in the development of MA has always been questioned. We herein report two cases with chronic liver disease and iron deficiency who concomitantly developed MA and PN lesions. Pruritus was the preceding factor and both lesions were confined to scratched areas. The association of two otherwise uncommon dermatoses in pruritic patients and their characteristic distribution might indicate an important role for pruritus-induced scratching in the pathogenesis of MA, too. PMID- 10846259 TI - Angiosarcoma of the scalp and face: failure of an interferon alpha treatment. AB - Angiosarcoma of the scalp and face is a rare malignant endothelial tumor arising in elderly people. Treatment is disappointing and prognosis remains poor. We report two cases of angiosarcoma of the scalp and face relapsing after classical therapy with surgery and radiotherapy and treated with interferon alpha. The tolerance was poor and the disease progressed 2 and 5 months after the onset of the treatment. When used alone, interferon alpha does not appear as an effective treatment in this kind of angiosarcoma. PMID- 10846260 TI - Recurrent follicular and lichenoid papules of sarcoidosis. AB - We describe a 62-year-old Japanese woman who exhibited recurrent follicular lichenoid lesions of sarcoidosis. The skin lesions appeared all over the surface of the body without any other organ involvement, and spontaneously regressed within 1 year. Two years later, the lesions recurred on the trunk, especially in the intertriginous areas such as the submammary areas and in fatty folds of the abdomen, and on the thighs and upper arms, with the appearance of uveitis of the right eye. Histological examinations showed that small epithelioid cell granulomas were localized in the perifollicular areas. The present case was characterized by a unique distribution of follicular lichenoid eruptions, which are rare manifestations of sarcoidosis and recurrence of the lesions. PMID- 10846261 TI - Diffuse large B-cell lymphoma associated with skin, muscle and cranial nerve involvement. AB - The present case, a 75-year-old man with extranodal B-cell lymphoma showed facial hemiplegia, paresthesia and cutaneous manifestations. He was initially diagnosed as having a facial paralysis of unknown etiology. One month after the original diagnosis, erythematous indurated plaques developed on his left cheek and nose. A skin biopsy from the plaque on his cheek showed dense infiltrates of large lymphocytes with irregularly shaped nuclei and prominent nucleoli in the dermis and subcutaneous tissue. The lymphocytes were positive for L26 and CD79a. A diagnosis of diffuse large B-cell lymphoma was made. A muscle biopsy from facial muscle in the area of the erythematous plaque showed massive destruction of the muscle tissues by the lymphomatous infiltrates. Furthermore, electrodiagnostic study showed peripheral cranial nerve palsies, involving the left facial and trigeminal nerves. We conclude that diffuse large B-cell lymphoma may develop symptoms such as facial hemiplegia and paresthesia prior to cutaneous manifestations. Diffuse large B-cell lymphoma must be considered as one of the important causes of palsies of cranial nerves at the peripheral level. PMID- 10846262 TI - An unusual cutaneous T cell lymphoma presenting as leonine facies. AB - We report an unusual case of a cutaneous T cell lymphoma with the clinical picture of leonine facies as the only skin symptom appearing during the first years of the disease. Small atypical lymphocytes with partly pleomorphic, partly indented cerebriform nuclei are present in the facial skin as well as in the peripheral blood and in the bone marrow. The lymphoma shows an indolent clinical behaviour without rapid progression of the disease. It shares features with both pleomorphic small/medium-sized T cell lymphoma and mycosis fungoides, but cannot be classified according to the criteria of the European Organization for Research and Treatment of Cancer (EORTC) Classification for Primary Cutaneous Lymphomas. At present the lymphoma has to be ranged among the group of so-called unspecified peripheral T cell lymphomas according to the proposal of the International Lymphoma Study Group. PMID- 10846264 TI - Manuel francisco allende AB - It was on the occasion of my first visit to San Francisco that I got to know Manuel Frank Allende. He was born in Uruguay in 1918 but spent time in Paris where he worked as a hospital registrar shortly before the war. In 1940, he saw the occupation of France by the Germans. He passed over the Spanish border, was briefly interned and then set off back to the United States. He had a brief "stay" at Ellis Island opposite Manhattan, before obtaining a resident's permit enabling him to continue his speciality, dermatology. He then left for California, his wife, Katharine's home state. Manuel Frank was very open-minded, benefiting from the Spanish, French and American cultures, a "man of the world" before the term had been invented. We soon got on well, he liked skiing, which he had practised in Alpe d'Huez before the war. I visited him in San Francisco where he lived in a house dominating the Bay near the Golden Gate. He drew and painted the wonderful view he had from his windows. He died suddenly on the slopes in the Rocky Mountains. Katharine still lives in San Francisco where she has long been a member of the governing body of the French hospital. Manuel Frank left a marvellous collection of drawings which illustrate well the kind of man he was. PMID- 10846263 TI - Current understanding of androgenetic alopecia. Part I: etiopathogenesis. AB - Androgenetic alopecia (AGA) is the most common type of hair loss in men and women. This continuous process results in a type of alopecia that follows a definite pattern in those individuals who are genetically predisposed. At present the predisposing genes are unknown but the relatively strong concordance of the degree of baldness in fathers and sons is not consistent with a simple Mendelian trait and a polygenic basis is therefore most likely. AGA can be defined as a DHT dependent process with continuous miniaturization of sensitive HF. Today we do not understand the molecular steps involved in androgen-dependent beard growth versus androgen-dependent hair loss in AGA. However, recent experimental and clinical advances enable us to explain some pathogenetic steps leading to androgenetic hair loss. Among other steroidogenic isoenzymes such as 17b- and 3b- hydroxysteroid dehydrogenases, the type 2 5a-reductase within the dermal papilla plays a central role by the intrafollicular conversion of T to DHT. PMID- 10846265 TI - Epidemiology of dental trauma: a review of the literature. AB - This article reviews recent reports describing the aetiology of dental trauma from national and international studies as well as the different classifications currently used to report dental injuries. It also discusses possible preventive measures to reduce the increasing frequency of dental trauma. Reported studies demonstrate that males tend to experience more dental trauma in the permanent dentition than females, however, there does not appear to be a difference between the sexes in the primary dentition. Accidents within and around the home were the major sources of injury to the primary dentition, while accidents at home and school accounted for most of the injuries to the permanent dentition. The most frequent type of injury was a simple crown fracture of the maxillary central incisors in the permanent dentition while injuries to the periodontal tissues were more common in the primary dentition. When preventive measures are being planned, knowledge of aetiology is important. More prospective studies from representative populations are required to understand the complexities of dental trauma epidemiology and to allow implementation of preventive strategies to reduce the increasing frequency of dental trauma. PMID- 10846266 TI - Assessing caries risk in children. AB - With the dramatic improvements in the oral health of children in Australia that have occurred over the past two decades, the option of, and the need for, targeted prevention of dental caries for those at high risk has become apparent. Since caries is of multifactorial aetiology, the clinical outcome varies depending on which factor, or combination of factors, is prominent in a particular individual; this may be related to both age and stage of life. Tests for caries risk can assist in prediction, but clinical signs and history are as important in assessing the main cause(s) of caries in an individual. In studies involving several factors, past caries experience (especially of the first permanent molar) continues to be the best predictor of future caries in children. Despite their ready availability, tests in the form of commercial kits are still expensive; no one test is an adequate predictor of caries risk, and the specificity and sensitivity of the tests are not reliably diagnostic for an individual. There is a need for regional longitudinal risk assessment studies in which potential risk factors are identified before the onset of caries in order to maximize predictive power and then validated against subsequent caries. Caries activity may not be able to be predicted in a population with low disease prevalence. Any risk assessment strategy must be followed by appropriate preventive interventions. PMID- 10846267 TI - Indigenous Australian dental health: a brief review of caries experience. AB - The indigenous community in Australia is an at risk population for oral diseases such as dental caries. The majority of communities are isolated and dental services in these areas are limited. Oral hygiene standards are poor and this combined with a diet rich in refined carbohydrates has led to high incidences of dental caries. In addition, diabetes, which is related to obesity (and a diet high in sugar and fat) has been linked to increases in oral disease. Caries prevalence was found to be low in areas where fluoridation levels in the water were high. The fact that the fluoride supplementation appears to improve oral health to a significant degree suggests that implementation of fluoride treatment programmes for school children and, where viable, fluoridation of water sources would be appropriate. In addition, dental education programmes should receive high priority. As with the rest of the community, these preventive measures will result in less need for emergency dental treatment in the future, better oral health for the community and reduced financial burden on the State. It is under these circumstances that oral health planners and providers must, in consultation with the relevant community representatives, develop appropriate mechanisms to address the needs of this group. The development of strategies that integrate with the plethora of general health strategies currently being implemented is just one means of achieving improved oral health outcomes for indigenous Australians. PMID- 10846268 TI - The effect on energy absorption of hard inserts in laminated EVA mouthguards. AB - One of the suggestions for using laminated mouthguards is the inclusion of hard inserts to improve mouthguard performance. However, there is a paucity of published material on the use of such inserts and this study was designed to investigate this theory. Hard layers of ethylene vinyl acetate (EVA) were included in laminated mouthguard sheets which were then subject to repeated impacts with an impact rig. Hard inserts resulted in reduced energy absorption when compared with a control sheet of the same material and approximate thickness but without the hard inserts. Additionally, the further the hard inserts were located from the impact surface, the least reduction there was on energy absorption. PMID- 10846269 TI - Studies of dental root surface caries. 1: Comparison of natural and artificial root caries lesions. AB - Qualitative polarized light microscopy and cross-sectional microhardness testing profiles were used to compare artificial root caries lesions with those naturally present in extracted human teeth. The artificial lesions were produced by immersion of roots with exposed windows in chemical buffers of acetate and lactate with no added enzymes or bacteria. The three buffer systems used produced lesions with the range of histological characteristics of natural lesions. These included a 'body' or 'surface' zone of the lesion when viewed in water, and a 'frontal' zone of reverse birefringence when viewed in quinoline. The 'frontal' zone was noted to first develop after three days of immersion in acetate buffer. Advanced natural lesions exhibited a wide, diffuse, advancing front, which was similar to that produced by 28-60 day lesions in acetate. Microhardness profiles of natural lesions could all be matched by one or more of the artificial lesions. The demineralizing process was seen to reduce mineral content across the lesion from 45 per cent in normal dentine to between 15 and 25 per cent. PMID- 10846270 TI - A prospective study of cantilever resin-bonded bridges: an initial report. AB - A prospective study was undertaken in a dental teaching hospital to compare the clinical performance of cantilever resin-bonded bridges with a fixed-fixed design for the replacement of a maxillary permanent incisor. A total of 25 anterior resin-bonded bridges was placed in 24 patients. The prostheses were evaluated over 14 to 45 months. During the evaluation period only one fixed-fixed type debonded, which was subsequently rebonded successfully. There was no debond in the cantilever group. The short-term, initial findings showed that cantilever resin-bonded bridges performed as successfully as their fixed-fixed counterparts and were considered to be an acceptable form of restoration. PMID- 10846271 TI - Caries experience among publicly-funded dental patients in Australia, 1995-96: type of care and geographic location. AB - Health card holders are a financially disadvantaged group and are the target population eligible for publicly-funded dental care. While their health status is generally worse compared with other Australians, there is also considerable variation among card holders. The aims of this study were to describe the oral health status of publicly-funded dental patients by type of care, geographic location and age, and to compare trends over time against other Australian studies. Patients were sampled randomly, based on date of birth, by State/Territory dental services in 1995-96. Dentists recorded oral health measures at the initial visit of a course of care using written instructions, but there was no formal calibration. The 6109 patients sampled were weighted in proportion to the numbers of publicly-funded dental patients for each State/Territory. Multiple linear regression analysis indicated that caries experience measured by the DMFT index increased across older age groups (p < 0.05). For rural compared with urban patients, mean numbers of decayed and filled teeth tended to be higher. For emergency compared with non-emergency care, mean numbers of decayed and missing teeth were higher, and filled teeth lower. The findings of this monitoring survey document high levels of previous disease and treatment and indicate variation between subgroups of users of publicly-funded dental care. This included an uneven geographic distribution of oral health and disease, and variation in unmet treatment needs by type of course of care. Temporal comparisons indicate publicly-funded patients have experienced the population trend towards lower levels of tooth loss over time but have higher levels of untreated decayed teeth compared with the general population. PMID- 10846272 TI - Maximizing response rates to a survey of dentists: a randomized trial. AB - Dentists currently practising within the Central Sydney Area were surveyed (n = 179). A randomized controlled trial was conducted to evaluate the effect of an advance telephone prompt (intervention group) compared with an advance letter prompt only (control group) in maximizing the response rate to a self administered questionnaire. While the overall response rate was 83 per cent, the final response rate (89 per cent) from dentists in the intervention group was significantly higher than that from dentists in the control group (78 per cent) (chi 2 = 4.14, df = 1, p = 0.04). Advance telephone prompts are effective in maximizing the response rates from dentists and are recommended in future surveys of this professional group. PMID- 10846273 TI - Facial and perioral molluscum contagiosum as a manifestation of HIV infection. AB - Molluscum contagiosum (MC) is a self-limiting viral disease of the skin and the mucous membranes. Facial and perioral MC is seen with increasing frequency in human immunodeficiency (HIV) infection, particularly in HIV infected homosexual men. The purpose of this study was to describe clinical observations of facial and perioral MC in HIV infected patients. One hundred and eighty HIV-positive individuals (160 males and 20 females) were examined over a period of five years. Fifty-eight were homosexual men and 19 were bisexual men. Fifty-one of 180 patients at the time of the first examination had CD+4 count < 200 cells/mm3 and another 63 presented loss of CD+4 cells in this level, during this period. Three HIV infected patients (two homosexual and one bisexual) were affected with facial and perioral MC. At the time of MC diagnosis the CD+4 count was less than 200 cells/mm3 for all three patients. One patient died nine months after MC diagnosis and the other two are still alive. It is remarkable that in this study no clinical lesions were observed on other sites of the skin. PMID- 10846274 TI - A cautionary tale. Case report. AB - A healthy 26 year old female underwent unsuccessful surgery for an incorrectly diagnosed sebaceous cyst in the cheek. Eventually the problem was traced to a non vital tooth 24. When the tooth was treated endodontically, the situation resolved in one week. PMID- 10846275 TI - Surgery combined with copper wire implantation in the management of cavernous orofacial haemangiomas. AB - This paper describes the management of a case of an extensive orofacial haemangioma by surgery combined with copper wire implantation. This technique of copper wire implantation in the management of orofacial haemangiomas was carried out for the first time in Jamaica by the authors. The use of percutaneous copper needles to induce therapeutic coagulation in cavernous haemangiomas, however, is not new, and this has been the basis of the use of copper wire. PMID- 10846276 TI - Temporomandibular joint arthroscopy: authors' reply. PMID- 10846277 TI - Temporomandibular joint arthrocentesis versus arthroscopy. PMID- 10846278 TI - Role of Australian dental schools. PMID- 10846279 TI - [Smoking cessation]. PMID- 10846281 TI - Cerebral thrombosis and vasculitis: an uncommon complication of ulcerative colitis. AB - Cerebral thrombotic disease is a rare and nearly always fatal complication of ulcerative colitis. It is associated with a necrotizing vasculitis. We report a fatal case with a confusing neurologic picture arising from this complication. Autopsy revealed necrosis and hemorrhages affecting both cortical grey and white matter. Microscopic examination showed thrombosis of small and medium size vessels associated with hemorrhages and a necrotizing angiitis. Ulcerations, hemorrhages, pseudopolyps, and cryptic abscesses were found in the rectosigmoid region of the colon compatible with active ulcerative colitis. A sudden neurologic deficit in a patient with ulcerative colitis should direct attention to the consideration of a cerebral thrombotic event and the possibility of an associated cerebral vasculitis. Diagnosis may be strongly suggested by MRI or arteriography, but it may require confirmation by biopsy of the brain parenchyma and leptomeninges. A hypercoagulable state has been associated with the thrombosis. Anticoagulation has yielded successful results in some patients with cerebral thrombosis but the risk of massive intracranial and gastrointestinal bleeding preclude to establish clear indications. Neurologic improvement has been obtained with the use of steroids and cyclophosphamide. PMID- 10846280 TI - Primary and secondary prevention of cardiovascular disease in the elderly. AB - Cardiovascular disease is the commonest cause of death and hospitalization in patients 65 years of age or older. The main offenders among the cardiovascular disorders in this age are CHD and hypertension. Many non-pharmacologic and pharmacologic measures in the middle-aged persons have confirmed for many years the effectiveness of primary and secondary prevention. Multiple intervention trials in the recent years have also demonstrated the effectiveness of these measures in patients 65 years of age and older. A summary of the main primary and secondary non-pharmacologic and pharmacologic measures that have been proven to be effective and useful in elderly patients has been presented with particular attention to hypertension and CHD. It also has been demonstrated that elderly patients have the capacity to follow the instructions of their physicians and that, as younger patients, they respond to these measures. PMID- 10846282 TI - Simultaneous occurrence of Hodgkin's disease and chronic lymphocyte leukemia: a unique presentation. AB - Chronic lymphocytic leukemia (CLL) is a chronic, low-grade hematologic malignancy that can transform to a large cell non-Hodgkin's lymphoma (Richter's syndrome), which is associated with an unfavorable prognosis. A distinct Hodgkin's disease subgroup of lymphomatous CLL transformation has been well characterized. We describe a patient presenting with simultaneous manifestations of CLL and Hodgkin's disease. This patient is unique, presenting with separate sites of involvement of each disease within the same organ, in this case the bone marrow. The morphologic and immunohistochemical findings at diagnosis are correlated with the findings of the postmortem examination. PMID- 10846283 TI - [Smoking cessation]. PMID- 10846284 TI - Attitudes toward euthanasia, assisted suicide and termination of life-sustaining treatment of Puerto Rican medical students, medical residents, and faculty. AB - OBJECTIVE: To elicit the opinion of Puerto Rican medical students, residents and internal medicine faculty as to the appropriateness of euthanasia and physician assisted suicide and end-of-life management. DESIGN: Survey using a 16-item questionnaire answered within a two-month period in the fall of 1996. SETTING: Rounds or faculty meetings at teaching hospitals located in the north, south and southwest of the island of Puerto Rico. PARTICIPANTS: There were 424 participants. The questionnaires of 279 medical students, 75 medical residents, and 35 internal medicine faculty members were analyzed. Thirty-five questionnaires, which were incomplete or answered by non-Puerto Rican participants, were excluded. MAIN OUTCOMES MEASURES: Frequency of support of active euthanasia, physician-assisted suicide, withholding or withdrawing life sustaining treatment with informed consent was determined. Whether it was ethical to prescribe full doses of drugs needed to alleviate pain even if it would hasten death, or agree to limit or restrict resources for the terminally ill was also determined. RESULTS: Forty per cent of the students, 33% of the residents, and 20% of the faculty supported euthanasia. If physician-assisted suicide were legalized, 50 per cent of the students, 43 per cent of the residents and 45 percent of the faculty would not be opposed to it. Sixty-eight per cent of the students, 67 per cent of the residents and 88 per cent of the faculty would support withholding or withdrawing life-sustaining treatment for dying patients with informed consent. Seventy-nine per cent of residents, 80 per cent of the faculty but only 54 per cent of medical students would prescribe full doses of drugs needed to alleviate pain in dying patients even if they would hasten death. Thirty-six per cent of the residents and faculty would agree to limit the use of medical resources for the terminally ill but only sixteen per cent of medical students would do so. CONCLUSIONS: The acceptance of euthanasia was inversely proportional to the clinical experience of the respondents: 40 per cent among students but only 20 per cent by the faculty. Withholding and withdrawing of life sustaining treatment was most acceptable to the faculty (88 per cent) but it was also favored by most of the students and residents (68 and 67 per cent respectively). Eighty per cent of the faculty, 79 per cent of the residents, but only 50 per cent of the students considered that prescribing full doses of drugs to alleviate pain if they knew it would hasten death, was ethical. The medical profession should take notice of evolving concepts in end-of-life management. PMID- 10846285 TI - Suicide, adolescents and Puerto Rico. AB - Suicide is a multifactoral phenomena. This article reviews the recent literature and attempts to identify those factors which have particular relevance for Puerto Rican adolescents. Risk factors that correlate highly with the Puerto Rican experience include homosexuality, due to the hostility that the person may experience, depression, gender, prevalence of psychiatric disorders, lack of social integration and social skills, military experience, cultural and religious factors, alcoholism, substance abuse and unemployment/poverty. The literature reviewed indicates that the Puerto Rican adolescent male is in a high risk group for suicide and that the risk increases with age, sexual preference, dysfunction in the family and substance abuse. PMID- 10846286 TI - Adolescent suicide: a review of the literature. AB - This article reviews the literature on the risk factors related to teen suicide in the United States and Puerto Rico. Findings indicate the interplay of multifactors including depression, homosexuality--due to the hostility that is often experienced by the person--, sexual abuse, lack of coping, social and problem-solving skills stemming from family dysfunction, feelings of isolation and helplessness, contagion, gender differences, alcohol and drug abuse, psychiatric disorders, biological factors, as well as natural disasters. Included in this report are some statistics on the prevalence of suicide among teens and in the military. PMID- 10846287 TI - [Determination of psychometric properties of the Geriatric Depression Scale (Yesavage & Brink, 1983) in a sample of aged Puerto Ricans]. PMID- 10846289 TI - Electrocardiogram of the month. Rosenbaum's syndrome. PMID- 10846288 TI - [Development of the Test of Attitudes and Knowledge concerning Old Age: a pilot project on its applicability in Puerto Rico]. PMID- 10846290 TI - Sentinel surveillance of campylobacter in England and Wales. PMID- 10846291 TI - Meningococcal disease associated with the Haj--update. PMID- 10846292 TI - The consequences of welfare reform for women's health: issues of concern for community health nursing. AB - Community health nurses confront the health effects of public policies on a daily basis, particularly with respect to low-income populations. Despite this, the effects of public policies are often attributed to problems that are more proximal in the environment than the broad-ranging scope of "public policy." This article argues the need for a broader perspective of environment that encompasses public policies, and provides an example of how specific aspects (time limits and the Medicaid eligibility process) of the Personal Responsibility and Work Opportunity Reconciliation Act (PRWORA) of 1996 may have harmful consequences for the health of low-income women. Finally, implications for community health nursing practice and the need for further nursing research are reviewed. PMID- 10846293 TI - Chart it once: innovation in public health documentation. AB - The Seattle-King County Department of Public Health implemented a revision in the charting for its Family Support programs with the goal of streamlining the entire charting process, reducing redundancy, improving quality, and improving productivity. This article applies diffusion of innovations theory to the process of changing documentation. PMID- 10846294 TI - Promoting helmet use among children. AB - Despite the proven efficacy of bicycle helmets, helmet use among children remains low. Helmet use among children riding bicycles significantly reduces head injuries and death. A variety of promotional programs to increase helmet use in children have demonstrated limited success. This article reviews a variety of approaches to increase helmet use among children. The PRECEDE-PROCEED model is applied and recommendations for community health nurses to promote helmet use among children are provided. PMID- 10846295 TI - Increasing access to prenatal care: an evaluation of minority health coalitions' early pregnancy project. AB - Early prenatal care utilization is a problem for racial and ethnic minority women in Indiana. Minority health coalitions in Indiana developed early pregnancy care coordination projects to address this need. To evaluate project outcomes, birth certificates of infants born to project mothers were matched with birth certificates of infants whose mothers did not receive care coordination. Results showed that project mothers were significantly more likely to start prenatal care earlier, had better prenatal care utilization as measured by the Kessner Index (Kessner, Singer, Kalk, & Schlesinger, 1973), and had better adequacy of care as measured by the APNCU Index (Kotelchuck, 1994) than the noncare-coordinated mothers. Although the project was successful in meeting its primary objective, specific recommendations are made for community health nurses to partner with the coalitions in improving the content of prenatal care, decreasing sociocultural barriers to care, and assisting in the evaluation of outcomes of future projects. PMID- 10846296 TI - Assessment of older urban drivers by nurse practitioners. AB - Thirty-five older urban adults who had forfeited a driver's license and 25 nurse practitioners (NPs) who provided health care to them participated in a qualitative study to (a) explore the perception of NPs regarding their role in assessing the driving abilities of older adults, (b) describe the ways in which NPs assist their clients to cope with the loss of a driver's license, and (c) delineate the NPs influence on the elders' decision to forfeit their license. Findings suggest that most NPs had concerns about their older clients' driving abilities. However, assessment of parameters related to driving safety was not their primary objective. An NPs recommendation to forfeit a license was important to the older driver's decision to do so. Assisting clients with the consequences of no longer driving was difficult for a variety of reasons. PMID- 10846297 TI - Health promotion and senior women with limited incomes. AB - Health promotion is increasingly being recognized as making an important contribution to the well-being of Canada's seniors. Most research relating to this topic, however, has focused on middle-income senior men and women. An exploratory study using ethnographic methods was conducted to explore and describe the health promotion experience of senior women living on limited incomes. Interviews with a total of 11 urban senior women living on limited incomes were analyzed. A major finding of this study was that the women utilized a wide variety of "ways of living" that are presented in the model, Health Promotion as Self Nurturance. Health promotion was perceived to be influenced by living on a limited income by most participants; however, 3 of the participants believed that their health status and income level were unrelated. Findings are discussed and implications for community health nurses are offered. PMID- 10846298 TI - Assessing child maltreatment. PMID- 10846299 TI - Bipolar disorder in ADHD children grown up. PMID- 10846300 TI - Inhalant dependence and withdrawal symptoms. PMID- 10846301 TI - Carbamazepine treatment of bipolar disorder in an adolescent with cerebral palsy. PMID- 10846302 TI - A review of tobacco smoking in adolescents: treatment implications. AB - OBJECTIVE: To review current data on the tobacco epidemic in adolescents that impact treatment decisions. METHOD: Epidemiological and pharmacological data, risk factors, characteristics of nicotine use in adolescents, and treatment intervention reports from the literature are discussed. RESULTS: Of students in grades 9 to 12, 42.7% have used tobacco; 75% of teenage smokers will smoke as adults. Environmental and biological factors influence adolescent smoking, including sociodevelopmental aspects of adolescence, psychiatric history, genetic background, ethnic and gender characteristics, drug effects, and regulatory factors. Criteria for nicotine dependence are currently based on the experience with adult smokers. Overall, smoking cessation treatment for adolescents has been disappointing because of low participation, high attrition, and low quit rates. CONCLUSION: Characterization of nicotine dependence and further assessment of the safety and efficacy of pharmacological treatment interventions in adolescents are needed given the formidable challenge of the tobacco epidemic in adolescents. PMID- 10846303 TI - A screening instrument for autism at 18 months of age: a 6-year follow-up study. AB - OBJECTIVES: A population of 16,235 children aged 18 months was screened using the Checklist for Autism in Toddlers (CHAT) to identify childhood autism (CA). Two further screening procedures were conducted at age 3 and 5 years. The population was followed up at age 7 years in order to establish the sensitivity, specificity, and positive predictive value of the instrument. METHOD: A brief checklist assessing joint attention and pretend play behaviors was administered by primary health care practitioners when the children were 18 months old. Follow up methods included screening through parents and health practitioners and checking medical and educational records. RESULTS: Nineteen cases of CA were successfully identified by the CHAT at 18 months. At follow-up a total of 50 cases of CA were identified via all surveillance methods. Thus, the CHAT has a sensitivity of 38% and a specificity of 98% for identifying CA. The positive predictive value of the instrument was maximized by concentration on the highest risk group. Repeated screening 1 month later increased the positive predictive value to 75% for identification of CA but reduced the sensitivity to 20%, although the specificity was close to 100%. The screen also identified cases of pervasive developmental disorder as well as children with language and other developmental disorders. CONCLUSIONS: The CHAT can be used to identify cases of autism and related pervasive developmental disorders at 18 months of age. It is emphasized that the CHAT is not a diagnostic instrument but can identify potential cases of autism spectrum disorders for a full diagnostic assessment. PMID- 10846304 TI - Attachment disorder behavior following early severe deprivation: extension and longitudinal follow-up. English and Romanian Adoptees Study Team. AB - OBJECTIVE: To examine attachment disturbances and disorder in a sample of children adopted into the U.K. following severe early privation and in a comparison sample of nondeprived, within-country, early-placed adoptees. METHOD: The subjects, 165 children adopted from Romania and 52 adoptees from the U.K., were assessed at age 6 years; longitudinal data (at ages 4 and 6 years) were available on the 111 Romanian adoptees placed in U.K. homes before 24 months of age and on all U.K. adoptees. Information on attachment disorder was derived from a semistructured interview with the parent; in addition, data on children's cognitive and social development were assessed using standardized assessments. RESULTS: Analyses revealed a close association between duration of deprivation and severity of attachment disorder behaviors. In addition, attachment disorder behaviors were correlated with attentional and conduct problems and cognitive level but nonetheless appeared to index a distinct set of symptoms/behaviors. Finally, there was marked stability in individual differences in attachment disorder behaviors and little evidence of a mean decrease over this 2-year period. CONCLUSIONS: The findings offer construct validation for the attachment disorder construct and highlight clinical and conceptual questions that require further research. PMID- 10846305 TI - Effect size of lithium, divalproex sodium, and carbamazepine in children and adolescents with bipolar disorder. AB - OBJECTIVE: To develop effect sizes for 3 mood stabilizers--lithium, divalproex sodium, and carbamazepine--for the acute-phase treatment of bipolar I or II disorder, mixed or manic episode, in children and adolescents aged 8 to 18 years. METHOD: Forty-two outpatients with a mean age of 11.4 years (20 with bipolar I disorder and 22 with bipolar II disorder) were randomly assigned to 6 weeks of open treatment with either lithium, divalproex sodium, or carbamazepine. The primary efficacy measures were the weekly Clinical Global Impression Improvement scores and the Young Mania Rating Scale (Y-MRS). RESULTS: Using a > or = 50% change from baseline to exit in the Y-MRS scores to define response, the effect size was 1.63 for divalproex sodium, 1.06 for lithium, and 1.00 for carbamazepine. Using this same response measure with the intent-to-treat sample, the response rates were as follows: sodium divalproex, 53%; lithium, 38%; and carbamazepine, 38% (chi 2(2) = 0.85, p = .60). All 3 mood stabilizers were well tolerated, and no serious adverse effects were seen. CONCLUSIONS: Divalproex sodium, lithium, and carbamazepine all showed a large effect size in the open treatment of children and adolescents with bipolar I or II disorder in a mixed or manic episode. PMID- 10846306 TI - Cognitive-behavioral group therapy for social phobia in female adolescents: results of a pilot study. AB - OBJECTIVES: To examine the efficacy of cognitive-behavioral group therapy for adolescents (CBGT-A) in females with social phobia and the effect of this treatment on the risk for major depression. METHOD: Female adolescents with social phobia (N = 35) were randomly assigned to treatment (n = 12) or no treatment (n = 23) groups. Assessments were conducted at baseline, after treatment, and at a 1-year follow-up. RESULTS: Eleven subjects completed treatment. Sixteen weeks of treatment produced a significant improvement in interference and reduction in symptoms of social anxiety. There was a significant reduction in the number of subjects meeting DSM-IV criteria for social phobia in the CBGT-A versus the untreated group; however, at the 1-year follow-up there were no significant differences by treatment condition. There was also suggestive evidence that treatment of social phobia lowers the risk for relapse of major depression among those with a history of major depression. Combining social phobia and major depression as the outcome produced more robust treatment effects in the 1-year follow-up. CONCLUSIONS: This pilot study provides evidence for a moderate short-term effect of CBGT-A for treating female adolescents suffering from social phobia and indicates that treatment of social phobia may result in a reduction of major depression. PMID- 10846307 TI - The experience journal: a computer-based intervention for families facing congenital heart disease. AB - OBJECTIVE: To test the feasibility and safety of a computer-based application designed to facilitate the healthy coping of children and their families who must contend with significant congenital heart disease (CHD). The application, called the Experience Journal (EJ), is a psychoeducational intervention based on a narrative model involving the sharing of personal stories about an illness. METHOD: Testing was conducted in 2 phases. In phase 1, 9 parents of children with CHD and 1 adult with CHD were asked to use the EJ. After utilization, semistructured interviews assessed EJ usability and safety. In phase 2, 40 mothers of children with CHD used the EJ during a hospitalization. Assessment of feasibility and safety was measured through the use of semistructured interviews prior to EJ utilization and 2 to 4 weeks after hospital discharge. RESULTS: Results revealed that the EJ was safe and useful for decreasing social isolation, increasing understanding of familial feelings about cardiac illness, and fostering positive reactions in mothers. CONCLUSIONS: Computer-based interventions that present psychoeducational and medical information closely connected to "one's own story" may open up new possibilities for families facing pediatric illnesses. PMID- 10846308 TI - The altering of reported experiences. AB - OBJECTIVES: The unreliability of human memory is well documented in the literature, yet psychiatrists and other mental health care professionals rely on patient self-report in history-taking. This study provides new evidence from a longitudinal study of autobiographical memory and discusses implications for the development and implementation of appropriate treatment plans and goals. METHOD: Seventy-three mentally healthy 14-year-old males were studied in 1962. Sixty seven of these subjects were reinterviewed face-to-face at age 48. Questions concerning areas of family relationships, home environment, dating, sexuality, religion, parental discipline, and general activities were asked in both interviews. RESULTS: Significant differences were found between adult memories of adolescence and what was actually reported during adolescence. Accurate memory was generally no better than expected by chance. CONCLUSIONS: If the accurate memory of one's past is not better than chance in the mentally healthy individual, even more care probably should be taken in obtaining accurate historical information in the medically, psychologically, or otherwise health compromised individual. It would be more constructive to treat recollections as existential reconstructions. PMID- 10846309 TI - Maternal characteristics and toddler temperament in infantile anorexia. AB - OBJECTIVE: To explore the association between specific maternal characteristics, maternal perceptions of toddler temperament, and infantile anorexia. METHOD: Three groups of toddlers (aged 12-37 months) participated in this study: toddlers with infantile anorexia (n = 34), picky eaters (n = 34), and healthy eaters (n = 34). Mothers completed questionnaires that assessed their own eating attitudes, marital satisfaction, and their toddlers' temperament, and an interview that explored their attachment representations. Mothers and toddlers were videotaped during a feeding session, and toddlers were weighed and measured. RESULTS: Temperament ratings differentiated between infantile anorexics and healthy eaters (p < .0001), with infantile anorexics receiving higher difficulty, irregularity, negativity, dependence, and unstoppable ratings. Mothers of anorexics showed greater attachment insecurity than mothers of healthy eaters (p < .05), but they demonstrated neither overt eating pathology nor less marital satisfaction than the other groups. Thirty-nine percent of variance in feeding conflict was explained by toddlers' diagnoses, temperament ratings, and maternal characteristics. Twenty-one percent of variance in toddlers' weights was explained by temperament ratings and feeding conflict. CONCLUSION: Maternal characteristics and perceptions of their toddlers' temperament characteristics should be addressed in treatment for infantile anorexia. PMID- 10846310 TI - Does actigraphy differentiate ADHD subtypes in a clinical research setting? AB - OBJECTIVE: To compare subtypes of attention-deficit/hyperactivity disorder (ADHD) (predominantly inattentive and combined types) and a comparison group on an objective measure of activity level (actigraphy). METHOD: Actigraphs were worn by 64 children (49 boys, 15 girls) during a full-day clinical diagnostic assessment; 20 subjects had a diagnosis of ADHD predominantly inattentive type, 22 had ADHD combined type, and 22 were non-ADHD controls. Mean actigraph scores were calculated for two 2-hour intervals, comprising, respectively, a psychometric evaluation in the morning and the completion of a speech and language assessment and research measures in the afternoon. RESULTS: There were no significant group differences in activity level in the morning session. During the afternoon session, children with ADHD were significantly more active than controls, but there were no differences between ADHD subtypes. CONCLUSIONS: These data partially support specifications in the DSM-IV regarding hyperactivity in ADHD; however, they also indicate that situational and/or temporal factors may affect the degree to which hyperactivity is expressed. Furthermore, the findings contradict specifications in the DSM-IV that suggest that children with ADHD combined type should be more hyperactive than children with ADHD predominantly inattentive type. PMID- 10846311 TI - Major depression and dysthymia in children and adolescents: discriminant validity and differential consequences in a community sample. AB - OBJECTIVES: To evaluate evidence, in a community sample, for discriminant validity between major depression (MDD) and dysthymia (Dy) in children and adolescents and to examine differential consequences of the 2 disorders for functioning. METHOD: The National Institute of Mental Health (NIMH) Methods for the Epidemiology of Child and Adolescent Mental Disorders (MECA) study consists of probability samples of youths. Data for this study are derived from interviews with 1,285 complete parent-youth pairs aged 9 to 17 years from 4 geographic areas in the United States. Youths with MDD were contrasted with those with Dy and those with both (MDD-Dy) on the NIMH Diagnostic Interview Schedule for Children, Non-Clinician Children's Global Assessment Scale, Columbia Impairment Scale, and the Service Utilization and Risk Factors Module. RESULTS: Groups with MDD, Dy, or MDD-Dy did not differ on sociodemographic, clinical, or family and life event variables. Youths with combined MDD-Dy were significantly less competent and more impaired than youths with either disorder alone. CONCLUSIONS: The findings do not provide support for the differentiation of MDD and Dy but strongly suggest the importance of addressing the needs of youths who meet criteria for both MDD and Dy because this combination is likely to be both serious and disruptive of normal developmental processes. PMID- 10846312 TI - Thought disorder in childhood schizophrenia: replication and update of concept. AB - OBJECTIVES: To examine whether measures of thought disorder differentiated schizophrenic from normal children and to examine the relationship of these measures with developmental and cognitive factors. METHOD: The speech samples of 88 schizophrenic and 190 normal children, aged 9 to 13 years, were coded with the Kiddie Formal Thought Disorder Rating Scale and Halliday and Hassan's analysis of cohesion. RESULTS: Above and beyond differences in mental age, gender, and neuroleptic status, the patients had significantly more formal thought disorder (FTD) and cohesive deficits than the normal children matched by mental age. The younger schizophrenic and normal children had significantly more thought disorder than the older children with these diagnoses. Combined FTD and cohesion scores correctly identified 76% of schizophrenic and 88% of normal children with little variability across mental age. The thought disorder measures generated 2 independent components: FTD and cohesion. CONCLUSIONS: Thought disorder measures that include both FTD and cohesion provide a quantitative diagnostic tool of childhood-onset schizophrenia. PMID- 10846313 TI - Childhood schizophrenia: responsiveness to questions during conversation. AB - OBJECTIVES: This study characterized further the communicative deficits associated with childhood-onset schizophrenia. It examined the use of speech functions that involve responses to Yes/No and Wh- questions in children with schizophrenia and normal children during conversation. It also ascertained the relationship of these speech functions with cognition and thought disorder. METHOD: Speech function variables, formal thought disorder, and cohesion were coded in 32 schizophrenic and 34 normal children, aged 5.6 to 12.4 years, from speech samples elicited with the Story Game. RESULTS: The schizophrenic children were significantly more impaired in the use of speech functions than the normal children. Other than the association of a subset of the speech functions with distractibility and loose associations, the speech function measures were unrelated to cognitive and thought disorder measures. CONCLUSIONS: Speech function analysis detects communication deficits not captured by thought disorder measures in children with schizophrenia. PMID- 10846314 TI - Case study: negative reinforcement and behavioral management of conversion disorder. AB - Behavioral interventions have been advocated for conversion disorder, but controlled trials are lacking. The authors report the case of a 12-year-old boy with conversion disorder after 3 months of persistent right arm pain and immobility whose symptoms rapidly resolved after an outpatient behavioral intervention using negative reinforcement. The importance of careful assessment, frank discussion of the diagnosis, patient and family psychoeducation, and a rehabilitative mindset are emphasized. Negative reinforcement may be a powerful tool in the management of pediatric conversion disorder, with the potential to reduce parental anxiety and prevent unnecessary physical assessments and interventions. PMID- 10846315 TI - Brief psychodynamic psychotherapy with young children. PMID- 10846316 TI - Quasi-experimental designs. PMID- 10846317 TI - Genetics of childhood disorders: XV. Prader-Willi syndrome: genes, brain, and behavior. PMID- 10846318 TI - Improving early options for abortion: it's time to stop waiting. PMID- 10846319 TI - The development of mifepristone: a pharmaceutical drama in three acts. AB - This paper reviews the various steps in development of mifepristone in Europe and in the United States. It is a personal account by the person who was in charge of its worldwide development and has participated in all the major decisions on the product. This paper presents a personal perspective on the development of mifepristone, explaining why this revolutionary medication is still not available in most of the world, including the United States. Briefly, the author's close and continuous involvement with the medical, regulatory, and commercial aspects of mifepristone development for more than 15 years provides a unique insider's view. The development of mifepristone has been so fraught with drama that I present it here as a theatrical piece. Nonetheless, the story has been told as objectively as possible. PMID- 10846320 TI - Mode of action of medical methods of abortion. AB - Several pharmacologic agents capable of inducing abortion have become available in the last 20 years. Prostaglandins cause powerful contractions of the uterus that lead eventually to expulsion of the fetal or embryonic tissue, but their effects on smooth muscle elsewhere in the body lead to such troublesome side effects as vomiting and diarrhea. Antimetabolites such as methotrexate inhibit DNA synthesis in rapidly dividing cells such as the trophoblast, resulting in release of prostaglandins and cytokines, extravasation of blood into the decidua, uterine contractions, and expulsion of embryonic or fetal tissue. Drugs that inhibit the synthesis of progesterone (epostane) or block its receptor (mifepristone) reverse the dominant influence of progesterone during pregnancy. As a result, a cascade of events is initiated, including influx of leukocytes and red blood cells into the decidua, release of prostaglandins and cytokines, and uterine contractions. Addition of uterotonic agents such as prostaglandins results in powerful uterine contractions, which supplement those induced by the withdrawal of progesterone. Because these methods reproduce many of the same physiological changes, clinical management of medical abortion is similar to that of spontaneous abortion. These methods provide a useful alternative to surgical abortion in early pregnancy. PMID- 10846321 TI - Medical abortion alternatives to mifepristone. AB - Social and political constraints have prohibited the use of mifepristone in most countries. As a result, researchers, clinicians, and women throughout the world have pushed for the development of alternative medical abortion regimens. Methotrexate (both oral and intramuscular) combined with misoprostol has been under investigation since 1993, with complete abortion rates similar to mifepristone regimens for pregnancies up to 49 days' gestation. Thousands of women have had safe medical abortions in their own homes using these medications. The overall abortion process using methotrexate and misoprostol takes longer, however, with 20% to 30% of women waiting up to five weeks for the abortion to occur. More recently, misoprostol-only regimens have been proposed for medical termination of pregnancy. The efficacy of these misoprostol regimens has been reported in the same range as those combining mifepristone or methotrexate with misoprostol, although other investigators have been unable to confirm these results. Additionally, a few small studies have tested tamoxifen combined with misoprostol, with mixed results. Continued research is necessary to ensure that reported efficacy rates for all proven medical abortion regimens are true for various populations of women. PMID- 10846322 TI - Research on regimens for early medical abortion. AB - Until recently the medical abortion regimen of mifepristone followed by a suitable prostaglandin analogue two days later has been available in only a few countries. Limited access to the drug has hampered research and forced investigators without access to seek alternatives. Where mifepristone has not been available, investigators have tested repeated doses of the prostaglandin misoprostol alone or in combination with methotrexate. However, these regimens cannot compete in efficacy, safety, and convenience with the mifepristone prostaglandin regimen. The development of the mifepristone-prostaglandin regimen has continued since approval, and a more effective and better tolerated medical method than was available ten years ago can now be offered to women. Several randomized double-blind studies have demonstrated that the dose of mifepristone can be lowered to 200 mg, one-third of the original dose, without compromising efficacy. Misoprostol has now largely replaced other prostaglandin analogues in the regimen, and this has made medical abortion simpler and more cost effective. Although women prefer oral administration of drugs, recent evidence suggests that to achieve sufficient efficacy, misoprostol has to be administered vaginally in women beyond 49 days' gestation. This medical abortion regimen has proved to be acceptable to women and safe when provided under proper conditions. PMID- 10846323 TI - A comparison of the Abortion Rights Mobilization and Population Council trials. AB - We compared the published results of the 1994-1995 Population Council (PC) trial to those from the 1996-1997 Abortion Rights Mobilization (ARM) trial to determine whether 200 mg mifepristone with 800 micrograms vaginal misoprostol is more effective and has fewer side effects than 600 mg mifepristone with 400 micrograms oral misoprostol. The complete medical abortion rate was lower in the PC trial than the ARM trial: 92% compared with 97% up to 49 days LMP (p < 0.05) and 83% versus 96% from 50 to 56 days LMP (p < 0.05). Nausea and vomiting were reported more frequently in the PC trial. The overall acceptability of the procedure was lower in the PC trial (88%) than in the ARM trial (94%), (p < 0.05). Mifepristone can be reduced from 600 to 200 mg when followed by vaginal misoprostol without loss of efficacy. Vaginal misoprostol extends the efficacy to 56 days LMP and is associated with less nausea and vomiting. Home use of misoprostol is safe and acceptable to women and decreases the number of required visits from three to two in most cases. PMID- 10846324 TI - Bleeding patterns after early abortion with mifepristone and misoprostol or manual vacuum aspiration. AB - OBJECTIVE: To prospectively determine patterns and predictors of bleeding after early abortion with mifepristone and misoprostol or manual vacuum aspiration (MVA). METHODS: Women fewer than 63 days from the last menstrual period and desiring termination of pregnancy were offered medical abortion with 200 mg oral mifepristone and 800 micrograms oral misoprostol or MVA under local anesthesia. Women completed a bleeding diary during the six weeks after abortion. Telephone calls were used to encourage diary completion and determine contraception use. RESULTS: 212 women were enrolled, 80% of whom completed diaries. Mean days of bleeding was higher in the medical (14 days) than MVA (9 days) group, but days of spotting (about 10) was similar in both groups. Increasing gestational age predicted more bleeding or spotting days after medical, but not MVA, abortion. Prolonged bleeding was not rare and was more common in the medical than the surgical abortion group. Combined oral contraceptive (COC) users and non-COC users had similar bleeding patterns after medical or surgical abortion. CONCLUSION: Bleeding after early medical abortion persists longer than after MVA and continues longer than previously appreciated regardless of method. Oral contraceptive use does not decrease bleeding after early medical or surgical abortion. PMID- 10846325 TI - Providing medical abortion: legal issues of relevance to providers. AB - As early medical abortion becomes more widely used and available in the United States, providers of women's health care are questioning whether, and in what way, existing abortion restrictions apply to medical abortion. Many of these laws, virtually all of which were written before early medical abortion was widely used in this country, make little sense in the context of medical abortion. Nonetheless, most abortion restrictions are broadly written and could be interpreted by state officials to apply to providers of medical abortion. This paper identifies and briefly describes common types of abortion restrictions, including physician-only laws, targeted regulation of abortion providers, fetal tissue examination and disposal laws, parental involvement requirements, and mandatory waiting periods, and explains the extent to which these types of requirements may be enforced against providers of medical abortion. In addition, because some abortion restrictions are irrational or impose significant and unwarranted burdens on women's access when applied to medical abortion, they may be vulnerable to legal challenge. We also review possible legal efforts to invalidate these laws, as well as legislative or regulatory changes that can be sought in order to make medical abortion truly accessible to women in this country. PMID- 10846326 TI - Medical abortion and the potential for new abortion providers: a cautionary tale. AB - The prochoice movement has expressed much hope that medical abortion will increase the pool of abortion providers because it does not require surgical training and does not need to be performed in specialized settings. By offering a detailed case study of the events surrounding a methotrexate abortion in a rural community health center by a family practice physician who had not previously performed surgical abortion, this article suggests that the diffusion of medical abortion to new providers and new settings may be more complex than some have suggested. The article illustrates that abortion is not only a medical procedure, but also a social process involving relations with large, often unfriendly bureaucracies as well as with the local community in which the abortion takes place. While still retaining optimism that medical abortion will ultimately increase the number of providers and hence increase access to abortion, the author believes that such a change will be considerably more gradual than commonly thought. PMID- 10846327 TI - Are US health professionals likely to prescribe mifepristone or methotrexate? AB - OBJECTIVE: To determine US health professionals' knowledge, attitudes, and practices regarding mifepristone and methotrexate as abortifacients. METHODS: In 1997, we conducted a nationally representative telephone survey of US obstetrician/gynecologists (OBs), family practice physicians (FPs), nurse practitioners (NPs), and physician assistants (PAs). RESULTS: Most health professionals surveyed considered themselves at least "somewhat familiar" with mifepristone or methotrexate, but few reported being "very familiar" with the methods. OBs were most likely to report being familiar with mifepristone (79%), followed by NPs/PAs (73%), and FPs (62%). FPs and NPs/PAs reported being less familiar with methotrexate than with mifepristone. If mifepristone were approved by the Food Drug Administration (FDA), 54% of OBs said they would be "very" or "somewhat" likely to prescribe the drug for abortion, including 35% of OBs who do not perform surgical abortions currently. About half of FPs and NPs/PAs were interested in offering mifepristone if approved by the FDA. Few health professionals reported ever having prescribed methotrexate for abortion. CONCLUSION: The providers most likely to express interest in offering medical alternatives were OBs who reported they "ever perform surgical abortions." Many health professionals who said they do not perform surgical abortions also said they would be interested in prescribing mifepristone, indicating a potential expansion of the number of health professionals who will provide abortion services. Medical protocol and legal issues may inhibit or slow expansion of the pool of providers offering medical abortion. PMID- 10846328 TI - Experiences and satisfaction with providing methotrexate-induced abortions among US providers. AB - OBJECTIVE: This study examined the acceptability of and experiences with methotrexate as an abortifacient among health care providers in the United States. METHODS: Seventy-six telephone interviews were conducted with providers (28% physicians, 13% midlevel providers, 29% administrators, and 30% counselors/other medical staff) of methotrexate-induced abortions during May and June of 1997. We examined provider satisfaction and several key operational issues relative to medical abortion, including: time spent with patients and staffing, training, and space needs. RESULTS: The majority of providers agreed that overall, more time was spent with methotrexate than with surgical abortion patients. Most agreed that the number of staff needed was the same for both methods, and that methotrexate required less office space. There were differing views on whether methotrexate required more, an equal amount of, or less training than surgical. Overall, providers were satisfied with methotrexate as an abortifacient. CONCLUSION: These data suggest that as knowledge of the method spreads, more providers will add this technique to their services. PMID- 10846329 TI - The counseling component of medical abortion. AB - OBJECTIVE: To investigate the specifics of counseling for medical abortion. METHODS: We used both self-administered surveys and semi-structured interviews to collect information from 77 medical abortion providers working in 56 sites. Providers were asked about staff training needs, the essential factors to be stressed in counseling, the time involved, and how counseling for medical abortion compares to counseling for surgical. RESULTS: Providers reported that counseling for medical abortion took longer than for surgical and that the time medical abortion takes to complete was the most important element to convey. The majority of respondents considered direct experience with patients to be the most effective form of training. CONCLUSION: This article presents some practical information and strategies for providers and clinic administrators interested in adding medical abortion to their services. PMID- 10846330 TI - Advanced practice clinicians and medical abortion: increasing access to care. AB - Early abortion is one of the safest and most common surgical procedures in the United States, yet it carries a significant stigma for both women and health care providers. Due to a number of factors, including political maneuvering and antichoice violence, the availability of physicians trained and willing to provide abortion has decreased dramatically over the past decades, thereby compounding the obstacles to early termination of pregnancy. At the same time, roles for nurse-midwives, nurse practitioners, and physician assistants have expanded in primary care as well as in specialty practice. Medical abortion is generating renewed interest in abortion care and is potentially more accessible to providers of primary care, women's health, and family planning services. Its provision by qualified advanced practice clinicians working with their physician colleagues promises to significantly improve the delivery of abortion services to women. PMID- 10846331 TI - In-depth interviews with medical abortion clients: thoughts on the method and home administration of misoprostol. AB - OBJECTIVE: To document women's experiences with a home-use mifepristone misoprostol medical abortion regimen. METHODS: We conducted in-depth interviews (n = 22) with women who received abortions in the New York City clinic of the ongoing Abortion Rights Mobilization clinical trial. Women received 200 micrograms mifepristone in the clinic and were randomized to self-administer 800 micrograms misoprostol vaginally at home either 24, 48, or 72 hours later. One week later, after their follow-up visits, women were asked to describe their experiences in their own words. RESULTS: The overwhelming majority of women found the home-use regimen acceptable. Nearly all said that they would not have preferred to return to the clinic for misoprostol, and many stated that the home use option was the principal appeal of medical abortion. Women described the medical procedure as "natural," private, and noninvasive, and, when describing their physical experiences, noted that the side effects were more tolerable in the comfort of their homes with someone familiar nearby for support. Additionally, most women felt prepared for the experience they encountered and competent in assessing any problems that arose. CONCLUSION: Home use of misoprostol affords women more flexibility, privacy, and control in their abortions, ultimately allowing them to seek their own comfort level. PMID- 10846332 TI - Instituting medical abortion services: changes in outcome and acceptability related to provider experience. AB - A prospective case series carried out at 34 Planned Parenthood sites studied the safety, efficacy, and acceptability of medical abortion with methotrexate and misoprostol in 1973 women. Women with pregnancies of 26 to 49 gestational days were given methotrexate followed by vaginal misoprostol. Eighty-one percent of women had documented complete medical abortions; abortion was not confirmed by examination in 6%, and 13% had documented suction curettage. The rate of suction curettage decreased with site experience, from 17% during the first 20 procedures to 10% at sites that had performed more than 50. Sites that had previous experience with either methotrexate or mifepristone medical abortion had a rate of 9% after they had performed at least 50 procedures. Exit interviews with 755 of the 902 women having abortions in the first year inquired about the level of comfort with the abortion and its overall acceptability. Women's satisfaction with the side effects did not directly correlate with site experience. Overall satisfaction with the abortion experience was related to whether the women had complete medical abortions or suction curettage. PMID- 10846333 TI - Is medical abortion acceptable to all American women: the impact of sociodemographic characteristics on the acceptability of mifepristone-misoprostol abortion. AB - OBJECTIVE: To determine whether mifepristone-misoprostol medical abortion is more acceptable to some American women than to others. METHODS: Using previously reported acceptability data from a large US multicenter study (n = 2121), we conducted a more detailed analysis to test whether characteristics such as race/ethnicity, education, age, gestational age, and geographic location affect acceptability. RESULTS: In the United States, Asian women were more than twice as likely as other women to choose this method because they believed it was safer, while white women were twice as likely to select it because they considered it more natural. More educated women were likely to select this method to show support for greater choice and were more pleased about being able to avoid surgery. Particular features of the protocol also appealed to different women. White and African-American women as well as women with more education would feel significantly more comfortable taking one or both of the drugs at home than they would in the clinic. Nearly all groups of women were equally satisfied with this method and found it highly acceptable. The data showed surprisingly few differences among women's overall satisfaction level, their willingness to choose the method again, or to recommend it to others. CONCLUSION: Mifepristone misoprostol medical abortion has widespread appeal to a broad range of women, but different women choose and prefer this method for different reasons. PMID- 10846334 TI - The knowledge, acceptability, and use of misoprostol for self-induced medical abortion in an urban US population. AB - OBJECTIVE: To assess the knowledge, acceptability, and use of misoprostol as an abortifacient in a primarily Latina population in the United States. METHODS: This was a cross-sectional study of 610 women who used one of three obstetrics/gynecology clinics in New York City. Participants answered a self administered questionnaire about their demographic and obstetrical characteristics; patterns of contraceptive use, including emergency contraception; general acceptability of abortion; perceptions of accessibility to abortion; prevalence of misoprostol use; and knowledge, attitudes, and availability of misoprostol. RESULTS: A substantial proportion (37%) of respondents admitted familiarity with the use of misoprostol as an abortifacient. The proportion of women who reported personal use of misoprostol (5%, 29/610) exceeded that reported in a large Brazilian prenatal care population (2.2%, 133/6102). Those who had taken misoprostol were more likely to have had abortions (p < 0.01), to be foreign born (p < 0.01), and to have known someone else who had used the drug (p < 0.01). Although the majority of women surveyed (73%) were Medicaid recipients, only half were aware that Medicaid covers abortion in New York State. CONCLUSION: The results of this descriptive study indicate a need to increase awareness of pre- and post-conception family planning methods and to remove barriers to access to these methods. Further research is essential to elucidate the knowledge and unsupervised use of misoprostol in other US populations. PMID- 10846335 TI - Opening a door to safe abortion: international perspectives on medical abortifacient use. AB - International experience compels us to revisit how we define and assess the safety and efficacy of medical abortifacients such as misoprostol. In some countries where safe abortion is neither accessible nor legal, even unsupervised, off-protocol use of misoprostol can provide women with a means to safely terminate pregnancy. This is due primarily to misoprostol-induced uterine contractions that cause bleeding, which in turn provides access to existing reasonable quality health services that would otherwise be unavailable. Several studies have suggested that an increase in the underground use of misoprostol in Brazil has already reduced serious complications from unsafe abortion. Thus, the availability of medical abortifacients combined with strengthened postabortion care services can legitimately be considered a public health success in countries in which safe abortion services do not exist and law reform is unlikely. PMID- 10846336 TI - Misoprostol alone--a new method of medical abortion? AB - Misoprostol shows promise for early termination of pregnancy. The drug is widely available, inexpensive, and easy to administer. Current evidence on the efficacy of potential regimens for early abortion, though, is hard to decipher and often contradictory. Research on a safe, effective, acceptable regimen of misoprostol alone for early abortion is underway. Available data show a wide range in efficacy of various regimens, and additional research is needed to understand this variability. A framework for additional research on misoprostol alone for medical abortion is presented with a focus on the need to evaluate benefits and risks of new regimens in the context of women's options for termination of pregnancy. We discuss the potential usefulness of a regimen with less efficacy than approved medical abortifacients. Research is needed as well on women's ability to self-diagnose incomplete abortion or ongoing pregnancy and the potential for misoprostol to be used as treatment for incomplete abortion in order to evaluate the benefits and risks of misoprostol-alone regimens. PMID- 10846337 TI - Early medical abortion in India: three studies and their implications for abortion services. AB - Although legal in India, abortion is frequently performed under unsafe or undesirable conditions. Moreover, the advancements required to make surgical abortion safe in India appear insurmountable in the near future. Because it requires a less extensive infrastructure than surgical abortion, medical abortion offers great potential for improving abortion access and safety now. To examine the feasibility of introducing medical abortion and to assess its potential as an alternative to surgical abortion, we conducted three separate studies on the use of 600 mg mifepristone and 400 micrograms oral misoprostol for medical abortion. Study 1 focused on the safety, efficacy, and feasibility of the standard French, three-visit protocol and was conducted in urban research centers in China, Cuba, and India. Study 2 liberalized the protocol to collect information from women using the method under more "real life" conditions in urban family planning clinics in India. Lastly, study 3 extended the trial to rural Indian villages to examine feasibility in settings typical of where the majority of the population resides. In all three settings in India mifepristone-misoprostol proved to be not only feasible, but safe and acceptable as well. With some changes to current protocols, medical abortion could now be safely phased into the existing health care infrastructure in India. Yet, medical abortion will bring its own set of service delivery challenges to address. PMID- 10846338 TI - Medical termination of early pregnancy: the Swedish experience. AB - Abortion has been legal up to the 22nd week of gestation in Sweden since 1975. Most women terminate their pregnancies early; 96% before the 12th week and 75% in less than 63 days. Termination of early pregnancy (up to 63 days' gestation) using mifepristone followed by gemeprost was approved in Sweden in 1992. Use of the medical method has slowly but steadily increased since then. Currently about 40% of the women eligible for the procedure choose it. Both the surgical (vacuum aspiration) and the medical methods are regarded as effective and safe. The medical method will not replace vacuum aspiration, but be an alternative to it. PMID- 10846339 TI - Medical abortion in China. AB - This paper reviews the literature on early medical abortion in China. Mifepristone was approved for termination of pregnancy up to 49 days' gestation in China in 1986. The commonly accepted regimens are 150 mg mifepristone administered in several small doses or a single 200 mg dose followed by 0.6 mg oral misoprostol. Complete abortion rates are 90% to 97%. Ultrasound is strongly recommended to rule out ectopic pregnancy. Patients remain in the clinic for four to six hours after misoprostol. If the fetal sac is not expelled during the four hour observation, the second dose of 0.4 mg misoprostol is given in many of the clinics. Rates of emergency curettage ranged from less than 1% to 4%, and rates of blood transfusion were less than 1%. The major complication of concern is undiagnosed ectopic pregnancy. The Chinese experience has shown that medical abortion is well accepted as an alternative to surgical intervention when provided under medical supervision and administrative monitoring. PMID- 10846340 TI - Abortion reporting in the era of medical procedures: why is it important? AB - Food and Drug Administration approval of mifepristone and the subsequent widespread use of medical abortion will change the patterns and practice of abortion services in this country. Accurate monitoring of new, nonsurgical abortion techniques will be critical as this change takes place. Providers will want to know which women will be the predominant users of medical versus surgical methods. If medical procedures are used widely, the national trend in gestational age at the time of abortion is likely to change because medical procedures are usually performed early (at less than eight weeks' gestation). New and existing abortion providers must be informed about the reporting statutes and requirements in their areas. Information and instructions for reporting abortions are available from the vital statistics offices in each state health department and the health departments of New York City and the District of Columbia. Ongoing comprehensive monitoring of legal induced abortion is needed in all states to determine the number of procedures performed, the characteristics of women who obtain them, and the evolving trends in procedures. PMID- 10846341 TI - There are just too many words.... PMID- 10846342 TI - Patient safety--the right prescription. PMID- 10846343 TI - Adequate physician documentation can help prevent payment errors. Doctors, hospital coders can partner to reduce mistakes. PMID- 10846344 TI - Diagnosis of ovarian vein thrombosis. PMID- 10846345 TI - The use of instant medical history in a rural clinic. Case study of the use of computers in an Arkansas physician's office. AB - This study evaluated the acceptance of using computers to take a medical history by rural Arkansas patients. Sex, age, race, education, previous computer experience and owning a computer were used as variables. Patients were asked a series of questions to rate their comfort level with using a computer to take their medical history. Comfort ratings ranged from 30 to 45, with a mean of 36.8 (SEM = 0.67). Neither sex, race, age, education, owning a personal computer, nor prior computer experience had a significant effect on the comfort rating. This study helps alleviate one of the concerns--patient acceptance--about the increasing use of computers in practicing medicine. PMID- 10846346 TI - Analysis of an SMN gene-like DNA fragment. AB - Part of a survival motor neuron (SMN) gene-like DNA fragment has been identified. This DNA fragment was accidentally isolated from cDNA by RT-PCR using primers specific for the region between exon 3 and 6 of the SMN gene. This fragment was used as a probe to hybridize the mRNA from several tissues, but we have been unable to detect any transcript of this SMN-like gene in these tissues. Thus, we have inferred this SMN gene-like fragment was a genomic product contaminant that was amplified in the reaction. Sequencing analysis of this fragment, which contains several stop codons, revealed a 74.6% nucleotide homology with the SMN gene. From these results, we believe that this DNA fragment is not a mutated form of SMN gene. Rather, it is an SMN-like pseudogene, which is variably present even in normal individuals. PMID- 10846347 TI - Monitoring of the levodopa concentration-response relationship in Parkinson's disease. AB - Motor fluctuations and abnormal involuntary movements are common complications encountered in advanced Parkinson's disease (PD) patients with long-term levodopa therapy. Monitoring of plasma levodopa concentrations and clinical effects has been reported to benefit the management of these complications. However, to our knowledge, there is no data available in Taiwan concerning the correlation between the plasma levodopa levels and motor fluctuations. In this study, we developed the laboratory methodology for plasma levodopa determination by using the aluminum extraction procedure and HPLC-ED. Serial blood samples and motor scores were obtained from 7 PD patients, and the correlation between plasma levodopa levels and motor responses were studied individually. In three patients with wearing-off phenomenon, plasma levodopa concentrations are compatible with the clinical "on" and "off" states. In the other four patients with complex fluctuating responses, their levodopa dosages were adjusted by the results of monitoring. Better motor responses were achieved by optimization of the levodopa pharmacokinetics in these patients. Our preliminary data suggest that simultaneous monitoring of plasma levodopa concentrations and clinical effects might be helpful to improve the therapeutic strategy in some of the parkinsonian patients with fluctuating responses to levodopa. PMID- 10846348 TI - Interleukin-8 secretion by cultured oral epidermoid carcinoma cells induced with nicotine and/or arecoline treatments. AB - Interleukin-8 (IL-8) is one of the multifunctional cytokines that can play a role on immune and inflammatory activities. Other in vitro observations indicated that IL-8 is a growth factor for keratinocytes. However, as the role of IL-8 in oral cancer cells is unclear, this study is thus designed to examine IL-8 secretion in cultured oral epidermoid carcinoma KB CCL17 cells treated with nicotine and/or arecoline. The cultures were treated with nicotine (1 or 100 microM) and arecoline (1 or 100 microM), alone or both, for 72 hrs. Enzyme-linked immunosorbent assay (ELISA) was used to examine IL-8 concentrations in culture supernatants. A repeated measure analysis of variance was used to identify differences among the treatments. Nicotine and arecoline, single or combined treatment, increased IL-8 secretion in KB CCL17 cells. When monoclonal 1 microgram/ml of antibody was added against IL-1 alpha or IL-1 beta in the treatment, IL-8 concentration significantly decreased compared with the non-added one. Exposure of cells to antibody against IL-1 alpha or IL-1 beta showed no significant increase in cell growth as compared with the control (medium alone). However, incubation of cells for 72 hrs in the presence of nicotine and/or arecoline treatments and antibody against IL-1 alpha or IL-1 beta significantly increased cell growth as compared with the antibody free one. It was concluded that IL-8 secretion by KB CCL17 cells may be partially mediated by IL-1 which could inhibit the KB CCL17 cell growth. Thus, IL-8 may be a vital participant in the cascade of interacting cytokines during smoking and areca quid chewing, inducing inflammation in oral cancer. PMID- 10846349 TI - The interference of uptake of thallium-201 in cultured rat myocardial cells with existence of potassium related pharmaceuticals--a preliminary report. AB - Thallium-201 myocardial perfusion imaging is wildly used to detect and assess the extent of jeopardized myocardial ischemia in the coronary artery disease and the viability of myocardium post infarction. In recent years, there has been a great deal of pharmacological development of blockers and openers of potassium channel. In this study, we will discuss the interference of uptake of thallium-201 ion in cultured neonatal rat myocytes with existence of a variety of pharmacological agents. The cultures of neonatal rat myocardial cells were incubated with different agents such as potassium chloride, sodium-potassium ATPase pump inhibitor (ouabain), cesium compound, variable potassium channel blockers (4 AP, TEA and glibenclamide) and their openers (minoxidil, and cromakalim). The radioactivity of intracellular thallium-201 that could enter rat myocardial cells was detected by gamma counter sixty minutes after thallium-201 was added. In this study we found that thallium and potassium ions behave in an analogous manner in cultured rat myocardial cells. Both 2.5 mM and 5 mM concentration of extracellular potassium ion significantly result in reduction of thallium-201 ion influx in rat myocardial cells. 0.5 mM ouabain, an inhibitor of sodium-potassium ATPase pump, reduced about 40% of influx of thallium-201 ion in cultured rat myocardial cells via active transport. Combination of both potassium ion and ouabain inhibit most of thallium-201 ions influx in myocardial cells, but it is not completely inhibited. Cesium, a potassium antagonist, also interferes with the uptake of thallium-201 in cultured rat myocytes in our study. The most interesting finding in our investigation is that potassium channel blockers such as TEA and glibenclamide, inhibit the influx of thallium-201 in myocytes. However, potassium channel openers have no overt effect on influx of thallium-201 in cultured rat myocytes. We indirectly observe about 60% of influx of thallium 201 ion into cultured rat myocardial cells via active sodium-potassium ATPase pump. Potassium, cesium and potassium channel blockers, such as TEA and glibenclamide, inhibited the different percentage of influx of thallium-201 in cultured rat myocardial cells in this study. PMID- 10846350 TI - An abnormal cholesterol profile in young adults with normocholesterolemic cerebral ischemia. AB - An abnormal cholesterol fraction can still be able to provoke cascades of lipidic atherogenesis even when the serum TC level is within normal range (< 200 mg%). However, there is a shortage of convincing data concerning cerebral atherogenesis in young Asians who have a different diet habit and living style from those in western countries. In this study, we examined the lipoprotein-cholesterol profile in young Taiwanese patients with noncardiac cerebral ischemia (NCCI) whose serum TC level was < 200 mg% and 200-250 mg%. The results showed a decrease of HDLC and an increase of VLDLC in patients with TC < 200 mg%, but only a decrease of HDLC in patients with TC = 200-250 mg%. The cholesterol fraction metabolism is obviously perplexed in NCCI subjects. These findings were not related to their associated risk factors. Accordingly, a derangement of cholesterol fraction with normal serum TC level can also incite lipidic cerebral atherogenesis in young Taiwanese adults. Therefore, a detailed evaluation of cholesterol profile should be born in mind in young eastern NCCI patients despite of a normal serum TC level. Tailored measure of diet and living should be modified to prevent lipidic atherogenesis in our society in future. PMID- 10846351 TI - Stressors of living with HIV/AIDS: patients' perspectives. AB - With the prospect of increasing numbers of patients with human immunodeficiency virus (HIV) and acquired immunodeficiency syndrome (AIDS), healthcare professionals (professionals) need to be better informed about patients' perspectives in order to provide efficacious, compassionate, and efficient care to patients. In Taiwan, the biomedical research and healthcare professionals' perspectives have dominated research related to HIV/AIDS. The purpose of the study was to investigate the stressors experienced by patients which have, thus far, been under-explored. This study used a phenomenological approach to gain insight into the patients' experience. In-depth, open-ended interviews were used to collect data. Interviews were tape recorded and transcribed verbatim to maintain data integrity and to reduce perceptual bias. The open coding procedure was used to analyze the interview transcripts. Study participants were 14 male patients with an average age of 35.71 year-old, who had known their positive HIV status for an average of 24.9 months. Patients experienced multiple stressors because of having the life threatening illness of HIV/AIDS. Their stressors were identified as follows: being confronted with the diagnosis, fear of disclosure, unpredictability of their physical condition, hypervigilance toward physical symptoms, emotional upheaval, and constrained life planning. The findings of this study form the basis for professionals to design services particularly addressed to patients' needs, for example, informed testing, and pre- and post-counseling. Further efforts could focus on developing systemic intervention programs to help patients better cope with the challenges of living with HIV/AIDS. PMID- 10846352 TI - A controlled study of postpartum depression in adult women. AB - The main purpose of the study was to examine the hypothesis that the postpartum period represents a time of increased risk for depression. The other psychosocial variables on stress, social support and self-esteem were also measured. Adult women, ranging in age from 22 to 45 years, comprised two samples: 148 postnatal women (22 to 44 years) and 148 controls (22 to 45 years). Five sets of instruments were used to collect data: the Demographic Data Form, the Perceived Stress Scale, the Interpersonal Support Evaluation List, the Coopersmith's Self Esteem Inventory, and the Beck Depression Inventory. Although the postnatal group tended to have a higher rate of depression, the difference was not statistically significant. No significant differences in stress, social support, self-esteem or depression were found between these two groups. However, postnatal women reported significantly higher somatic symptoms of depression than controls. Results of the stepwise multiple regression indicated that the best subset to predict postpartum depression was self-esteem, stress, postnatal complication and work status. The best subset to predict depression of controls included self-esteem, social support, socioeconomic status and stress. Our data indicate that the psychosocial health status of postnatal women is not significantly different from the controls, although the postnatal women complain more about the loss of bodily functions. The possible explanations deserve further research. PMID- 10846353 TI - Otogenic brain abscess--a case report. AB - Brain abscess is one of the life-threatening complications of otitis media. Mortality and morbidity have decreased with the advent of antibiotic therapy. More frequently encountered in cases of acute otitis media in the preantibiotic era, in recent years otogenic brain abscess was noticed almost only in patients of chronic otitis media with cholesteatoma. A case of brain abscess in a 49-year old female was initially diagnosed as a headache. A high resolution computed tomography (HRCT) scan of the temporal bones later revealed that there were two abscesses over the right side temporal lobe. A modified radical mastoidectomy was performed. Cultures of the middle ear cholesteatoma later grew Pseudomonas aeruginosa and Strenotrophomonas maltophilia. Antibiotic therapy was carried on for three months postoperatively. The patient improved but retained a conductive hearing loss. PMID- 10846354 TI - [Three-channeled aortic dissection: selection of surgery based on the images]. AB - Three patients with 3-channeled dissection were operated upon. Images of the dissection were enlargement of the false lumens, compression of the true lumen by enlarged false lumens and visceral arteries of false lumen origin. These prevent the use of cardiopulmonary bypass (CPB) and cause malperfusion of the viscera. Three-channeled dissecion is easy to rupture for its peculiar anatomy and total repair of the thoraco-abdominal aorta is mandatory. Fenestration brings functional recovery of malperfused viscera and enables the patients to be placed on CPB for total repair. Two patients underwent infrarenal and descending aorta fenestration followed by the total repair of thoraco-abdominal aorta successfully. A third patient has been placed on the strict CT follow-up following the infrarenal fenestration. PMID- 10846355 TI - [An autopsy case of thymic carcinoma producing various tumor markers and the examination of 222 autopsy cases of thymic malignant tumor in Japan]. AB - The autopsy of a 76-year-old Japanese female patient, which revealed thymic carcinoma with various tumor markers such as NSE, CYFRA, and CA-125, is presented. The patient died from hepatic failure because the liver was overtaken by the tumors. At autopsy, the thymic carcinoma was found to have metastased only in the liver. From microscopical analyses and electron microscopical findings, we diagnosed poorly differenciated squamous cell carcinoma of thymic origin. In the histochemical analyses, the tumor cells were positively stained in CA 125, CA 19 9, EMA, NSE, AE 1, AE 3, CEA, S-100, glimerius and Bcl-2. These date suggest that the tumor cells produced various tumor markers. In 222 autopsy cases of thymic malignant tumor observed in Japan over a period of 4 years, the dominant pathohistological image was squamous cell carcinoma. It is interesting that the greatest number of combined malignant tumors with thymic malignancies were thyroid papillary carcinomas. PMID- 10846356 TI - [A case of saccular aneurysm of the coronary artery post PTCA]. AB - We experienced a 48-year-old man who was diagnosed as having a saccular aneurym of the coronary artery by CAG after an otherwise uneventful percutaneous transluminal coronary angioplasty (PTCA). At the time of operation, the aneurysm, measured 15 mm, and arose from the 3rd segment of the right coronary artery. Aneurysmal formation after successful PTCA is not rare and is related to the overinflating balloon. It is necessary to evaluate the coronary artery after PTCA by IVUS. There were no symptoms after the operation and he was discharged at 21th postoperative day. PMID- 10846357 TI - [A case of repeated surgical intervention for pulmonary venous obstruction after repair of total anomalous pulmonary venous connection]. AB - A 3-month-old boy showed pulmonary venous obstruction after repair of total anomalous pulmonary venous connection. He was treated with a novel method of stureless in situ pericardium repair. Six weeks after this operation, he showed recurrence of venous obstruction and the same procedure was performed. Another six weeks after this operation, stenosis of the left pulmonary vein necessitated the same procedure again. Although he suffered from pneumonia and died of sepsis, this procedure revealed effectiveness for at least three months. This report showed clinical course after repeated stureless in situ pericardium repair. More experience will be necessary to evaluate the effectiveness of this procedure. PMID- 10846358 TI - [Surgical resection of lung cancer in a patient with von Willebrand's disease]. AB - Von Willebrand's disease (vWD) is one of the most common inherited hemorrhagic disorders, and there have been few reports on major thoracic surgery performed in a patient with the disease. We report a vWD patient who underwent of surgical resection of lung cancer. A 67-year-old gentleman, was diagnosed to have vWD when he was an infant, but he needed no medical treatment. Abnormal shadow in the lung was detected on a mass screening chest X-ray. We performed open biopsy, which proved the shadow to be large cell cancer, and performed right upper lobectomy and lymph node dissection. We used Contact-F, monitoring factor VIII, vWD factor and factor VIII-related antigen, during and after the operation, with a successful result. PMID- 10846359 TI - [Experimental study of calcification of the xenopericardia]. AB - We studied the calcification of four kinds of xenopericardium used in cardiac surgery. Group 1: bovine Pericardium (Tissueguard. Biovascular, inc), Group 2: bovine pericardium (No-React. Shelhigh, inc), Group 3: porcine pericardium (Rigg. Polystan, inc), Group 4: equine pericardium (Xenomedica. Baxter-Edwards, inc). Each pericardium was implanted in abdominal subcutaneous pouches of rats. The mean calcium content of each Group after eight weeks of subcutaneous implantation were 0.67 +/- 0.15 mg/g, 0.51 +/- 0.15 mg/g, 161 +/- 18 mg/g and 173 +/- 18 mg/g in Group 1, Group 2, Group 3 and Group 4, respectively. There was no significant difference between Group 1 and 2, or between Group 3 and 4. On the other hand, there were significant differences between Group 1, 2 and Group 3, 4 (p < 0.001). The deposition of calcium and inflammatory changes were markedly observed microscopically in Group 3 and 4, but in Group 1 and 2, they were only slightly observed. CONCLUSION: Two kinds of bovine pericardium are superior to both porcine and equine pericardium in the point of less calcium deposition and inflammatory cellular response after implantation. PMID- 10846360 TI - [One advice to the decision of graft size for ascending and total arch graft replacement (in a case of differential diameters in ascending aorta and descending aorta)]. AB - We carried out the surgery of thoracic aortic aneurysm in fifty-eight patients from June 1994 to February 1999 (including aortic dissection in twenty-six patients). The mean size of grafts were 28.1 mm in ascending graft replacement, 25.8 mm in both ascending and arch graft replacement and 23.8 mm in descending graft replacement. The grafts for ascending aortic aneurysm were significantly larger than those for descending aortic aneurysm. In two of twenty-six patients undergoing both ascending and arch graft replacement, different size of grafts were used for ascending replacement and for arch replacement with satisfactory results in terms of bleeding from the anastomotic sites. Case 1; A 45-year-female with aortitis syndrome and aortic regurgitation due to annuloaorticectasia and thoracic aortic aneurysm underwent simultaneous aortic root replacement with composite graft (25 mm St. Jude Medical valve and 28 mm Hemashield graft) and total arch replacement (30 mm Hemashield graft with two side branches). Case 2; A 64-year-female was diagnosed as chronic type II dissecting aneurysm combined with acute type I aortic dissection. Ascending aorta was replaced with a 26 mm Hemashield graft, and the aortic arch was replaced with a 24 mm Hemashield graft with three side branches. PMID- 10846362 TI - [The management of aortic root replacement using the Top-Hat/Gelweave composite graft]. AB - Composite graft replacement of the aortic root has become a routine procedure for annuloaortic ectasia (AAE) and aortic valve insufficiency (AR) with aortic dissection and the results have improved. We treated six cases of aortic root reconstruction using the Carrel patch method in 1998. The Top-Hat/Gelweave Composite graft fit together well and the procedure is technically similar to standard valve replacement. Upon measuring the valve size a Gelweave graft 1 mm larger than the valve size should be selected. There were no incidence of hemorrhage or postoperative hemolysis. Further long-term follow-up is necessary. PMID- 10846361 TI - [Open heart surgery in three patients receiving dialysis for more than 20 years]. AB - In Japan, the number of chronic hemodialysis (HD) patients is increasing, and there are 7,000 cases with a more than 20 year history of HD. At our institute, we have experienced 135 cases of open heart surgery in patients on HD, including 92 isolated CABG cases and 43 other open heart surgery cases. However, open heart surgery for patients with a more than 20 year history on HD is rare. Open heart surgery on HD patients is rather difficult, since perioperative management can be complicated and special care must be taken for prophylaxis of infection, fluid and electrolyte management and anticoagulation therapy. Many complications have been published in HD patients. At our institute, 3 cases of open heart surgery in patients with more than a 20 year history of HD have been performed. These included triple CABG, double CABG + AVR and double CABG in a post-kidney transplantation patient. They were discharged uneventfully with angiographically patent grafts. It is concluded that for CABG in patients on HD, aggressive use of arterial conduits for revascularization is recommended. For patients with a transplanted kidney, careful management against dysfunction and rejection is necessary. PMID- 10846363 TI - [Surgery for lung cancer in octogenarians]. AB - Twenty-five octogenarians with lung cancer underwent resection: 15 lobectomies, 3 segmentectomies, and 7 wedge resections. There were two operative deaths (8%) and one hospital death (4%). The complication rate was 26%. All complications occurred in cases with lobectomies. The cumulative 5-year survival rate was 47.4%, 57% for patients for curative and relative non-curative surgery, and 0% for absolute non-curative surgery. It is considered that limited operation for lung cancer in octogenarians is acceptable. PMID- 10846364 TI - [A case of chronic localized thoracoabdominal aortic dissecting aneurysm]. AB - A rare case of chronic localized thoracoabdominal aortic dissecting aneurysm was surgically treated. The patient was a 76-year-old woman. She was reffered to our hospital because of the abnormality of the chest roentogenogram. Chest CT and aortography revealed localized thoracoabdominal dissecting aneurysm 70 mm in diameter. Patch closure of intimal tear was carried out under partial cardiopulmonary femoro-femoral bypass. The patient discharged on the 23rd post operative day without any complications. PMID- 10846365 TI - [A surgical case of atrial septal defect with myotonic dystrophy]. AB - A case report is presented of a 16-year-old woman with myotonic dystrophy, who required open-heart surgery for an atrial septal defect. We successfully repaired an atrial septal defect using normothermic cardiopulmonary bypass and electrically maintained ventricular fibrillation. Anesthesia using fentanyl, vecronium and isoflurane was uneventful and there were no postoperative complications. Anesthetic agents must be selected with care, and ventilation requires meticulous attention. PMID- 10846366 TI - [A case of ascending aortic aneurysm with intrathoracic goiter]. AB - A case of the successful operation for ascending aortic aneurysm and intrathoracic goiter was described. A 71-year-old woman was admitted to our hospital for the treatment of ascending aortic aneurysm disclosed by the examination in another hospital. Preoperative chest CT showed ascending aortic aneurysm as large as 8 cm in diameter, and an upper mediastinal tumor compressing the main bronchus. The tumor was continuous with the right lobe of the thyroid. The excision of the tumor was performed through median sternotomy with cervical collar incison. After that, the replacement of the ascending aorta was carried out under cardiopulmonary bypass and hypothermic circulatory arrest. The histological examination of the resected specimen revealed adenomatous goiter without malignancy. PMID- 10846367 TI - [A case of successful reoperation for distal aortic arch pseudoaneurysm after replacement of descending aorta]. AB - The patient was a 69-year-old man who had undergone replacement of descending aorta under close clump technique. Hoarseness occurred 8 years after the operation. Computed tomography and aortography revealed a saccular aneurysm of the distal aortic arch. During surgery, the pseudoaneurysm originated from a intimal defect close to the proximal anastomotic site of the graft. Total aortic arch replacement was successfully performed under retrograde and selective cerebral perfusion. The postoperative course has been uneventful. PMID- 10846368 TI - [Modified extended septoplasty for left ventricular outflow tract obstruction after repair of double-outlet right ventricle report of a case]. AB - A three-year-old boy with residual VSD and LVOTO after repair of double outlet right ventricle underwent modified extended septoplasty successfully. Preoperative left ventriculography showed a residual VSD and LVOTO. The peak systolic pressure gradient between the left ventricle and aorta was 38 mmHg. Operation aimd not only at relief of the obstruction, but also at streamlining of the LVOT by removal of protrusions. We modified the extended septoplasty reported by Belli et al in 1996. The postoperative course was uneventful and patient was discharged at 15 postoperative day. The most recent echocargiogram revealed no residual LVOTO. PMID- 10846369 TI - [A case of pulmonary actinomycosis radiologically mimicking a lung cancer]. AB - A 59-year-old male clerk consulted in general practitioner due to cough and hemoptysis. A mass shadow was pointed out in the left upper lung field on a chest radiograph. Patient was referred to our hospital for further treatment. Any definitive daiagnosis could not be made after examinations including sputum culture, cytology and TBLB. Because a lung cancer was strongly suspected, an exploratory thoracotomy was performed. Actinomyces was detected by pathological study of excised specimen, with no evidence of cancer. ABPC was administered for two months postoperatively. The patient is doing well without recurrence of actinomycosis 2.5 years after the surgery. Pulmonary actinomycosis presenting a mass shadow on a radiograph may mimick a pulmonary tumor, especially a lung cancer. Pulmonary actinomycosis should be considered in a differential diagnosis of pulmonary lesion thought to be malignant. PMID- 10846370 TI - [A case of pulmonary adenocarcinoma of arising from the wall of a giant bulla which disappeared spontaneously]. AB - In a 53-year-old man, giant bulla was detected on the left side on chest X-ray and mass shadow was demonstrated near the bullae (20 x 10 mm). CT scan did not show a mass shadow and the shadow was diagnosed as scar formed by the bulla wall. Three months later, the bulla had disappeared spontaneously, but an abnormal shadow at the apex of left lung was pointed out. Chest CT findings suggested lung cancer arising from the wall of the collapsed giant bullae and partial resection of the left upper lobe was performed. The postoperative pathological diagnosis was moderately differentiated adenocarcicoma (12 x 5 x 5 mm) arising from pulmonary bullae. It must be noted that occult cancer may exsist at the wall of giant bullae, the surgical procedure involving complete resection of the bullae wall must be selected. PMID- 10846371 TI - [A surgical case of adult Bochdalek hernia assisted by thoracoscopic surgery]. AB - A case of adult Bochdalek hernia in a naturally healthy 49-year-old woman is described. She was seen at our hospital because of cough lasting for about one month. The chest X-ray film revealed a gastrointestinal gas image in the left thoracic cavity. In addition, the CT scan and upper gastrointestinal series showed prolapse of the stomach, small intestine and a part of transverse colon. Operation was performed through a mini-thoracotomy using a thoracoscope assistedly. No hernia sac was observed and the diaphragmatic defect was 6 x 5 cm in size. After the organs was restored, the defect was closed with continuous over-and-over suture and the diaphragm was enhanced by PROLEN MESH. PMID- 10846372 TI - Violence risk assessment and risk communication: the effects of using actual cases, providing instruction, and employing probability versus frequency formats. AB - This article describes studies designed to inform policy makers and practitioners about factors influencing the validity of violence risk assessment and risk communication. Forensic psychologists and psychiatrists were shown case summaries of patients hospitalized with mental disorder and were asked to judge the likelihood that the patient would harm someone within six months after discharge from the hospital. They also judged whether the patient posed a high risk, medium risk, or low risk of harming someone after discharge. Studies 1 and 2 replicated, with real case summaries as stimuli, the response-scale effects found by Slovic and Monahan (1995). Providing clinicians with response scales allowing more discriminability among smaller probabilities led patients to be judged as posing lower probabilities of committing harmful acts. This format effect was not eliminated by having clinicians judge relative frequencies rather than probabilities or by providing them with instruction in how to make these types of judgments. In addition, frequency scales led to lower mean likelihood judgments than did probability scales, but, at any given level of likelihood, a patient was judged as posing higher risk if that likelihood was derived from a frequency scale (e.g., 10 out of 100) than if it was derived from a probability scale (e.g., 10%). Similarly, communicating a patient's dangerousness as a relative frequency (e.g., 2 out of 10) led to much higher perceived risk than did communicating a comparable probability (e.g., 20%). The different reactions to probability and frequency formats appear to be attributable to the more frightening images evoked by frequencies. Implications for risk assessment and risk communication are discussed. PMID- 10846373 TI - Perceived conditions of confinement: a national evaluation of juvenile boot camps and traditional facilities. AB - In a national study of juvenile correctional facilities, the perceived environment of 22 juvenile boot camps was compared to the perceived environment of 22 traditional facilities. Self-report surveys completed by 4,121 juveniles recorded information on demographics, risk factors, and perceptions of the facility's environment. Compared to juveniles in traditional correctional facilities, boot camp residents consistently perceived the environment as significantly more controlled, active, and structured, and as having less danger from other residents. Boot camp juveniles also perceived the environment as providing more therapeutic and transitional programming. Overall, from the perspective of the juveniles, boot camps appear to provide a more positive environment conducive to effective rehabilitation considering almost all of the conditions measured. A major concern is that in both types of facilities, juveniles perceived themselves to occasionally be in danger from staff (rated as rarely to sometimes). PMID- 10846374 TI - The law concerning the conduct of lineups in England and Wales: how well does it satisfy the recommendations of the American Psychology-Law Society? AB - Four rules minimize the likelihood of a false conviction resulting from the misidentification of a suspect from a lineup: (1) The person conducting the lineup should not know which member of the lineup is the suspect. (2) The eyewitness should be warned that the criminal might not be present. (3) Foils should be selected based on the eyewitness's verbal description of the criminal. (4) Confidence should be recorded at the time of identification. In this paper the relevant law relating to lineups in England and Wales is outlined and the extent to which they satisfy the four rules is reviewed. It is concluded that the way in which lineups are conducted in England and Wales would, with minor modifications, satisfy the four rules, and this demonstrates that the rules can be applied practically. PMID- 10846375 TI - Assessing pretrial publicity effects: integrating content analytic results. AB - When a case has received pretrial publicity which has the capacity to bias potential jurors in the trial venue, a change of venue is one means of attempting to ensure that the defendant receives a fair trial. Content analysis of the pretrial publicity surrounding a case can provide the court with important information to consider when determining whether prejudice in the relevant community is too great for the defendant to receive a fair trial. This paper presents an approach to content analysis of pretrial publicity that draws upon both legal commentary and past empirical social science research. It is a systematic approach that could be employed by both the prosecution and defense when presenting arguments to the court about whether a change of venue should be granted. Information gleaned from content analysis of the publicity surrounding a specific case fills the gap between information provided by experimental research which has examined pretrial publicity effects and public opinion polls concerning the public's perception of the defendant in a particular case. Results from a content analysis can serve to validate public opinion survey data gathered from the same locales. To exemplify this content analytic approach, a content analysis conducted by the authors in preparation for the change of venue hearing in the case of Timothy McVeigh is presented. PMID- 10846376 TI - Discrimination and instructional comprehension: guided discretion, racial bias, and the death penalty. AB - This study links two previously unrelated lines of research: the lack of comprehension of capital penalty-phase jury instructions and discriminatory death sentencing. Jury-eligible subjects were randomly assigned to view one of four versions of a simulated capital penalty trial in which the race of defendant (Black or White) and the race of victim (Black or White) were varied orthogonally. Dependent measures included a sentencing verdict (life without the possibility of parole or the death penalty), ratings of penalty phase evidence, and a test of instructional comprehension. Results indicated that instructional comprehension was poor overall and that, although Black defendants were treated only slightly more punitively than White defendants in general, discriminatory effects were concentrated among participants whose comprehension was poorest. In addition, the use of penalty phase evidence differed as a function of race of defendant and whether the participant sentenced the defendant to life or death. The study suggest that racially biased and capricious death sentencing may be in part caused or exacerbated by the inability to comprehend penalty phase instructions. PMID- 10846377 TI - Permitting jury discussions during trial: impact of the Arizona reform. AB - A field experiment tested the effect of an Arizona civil jury reform that allows jurors to discuss evidence among themselves during the trial. Judges, jurors, attorneys, and litigants completed questionnaires in trials randomly assigned to either a Trial Discussions condition, in which jurors were permitted to discuss the evidence during trial, or a No Discussions condition, in which jurors were prohibited from discussing the evidence during trial according to traditional admonitions. Judicial agreement with jury verdicts did not differ between conditions. Permitting jurors to discuss the evidence did affect the degree of certainty that jurors reported about their preferences at the start of jury deliberations, the level of conflict on the jury, and the likelihood of reaching unanimity. PMID- 10846378 TI - [Effects of chlorpromazine on the skeletal muscle--a study using skinned single fibers of the guinea pig]. AB - Chlorpromazine (CPZ) was reported to influence the functions of the sarcoplasmic reticulum (SR). To determine the action sites of CPZ on calcium functions in skeletal muscle, the calcium-induced calcium release (CICR) rates from SR, Ca2+ uptake rates into the SR, and Ca2+ sensitivity to the contractile system were examined using the single skinned fiber. CPZ enhanced slightly rates of CICR in a dose-dependent manner. The Ca2+ uptake rates into the SR were significantly attenuated at the concentrations of 10 and 30 microM of CPZ. CPZ in a concentration below 10 microM did not affect Ca2+ uptake rates. This agent did not enhance Ca2+ sensitivity of the contractile system. Although CPZ was suggested to increase Ca2+ release from the SR by Takagi, this observation might have resulted from either the increase of Ca2+ release or the inhibition of Ca2+ uptake since Ca2+ might be pumped up to the SR during the release of Ca2+ in his experimental conditions. Endo's method enabled us to analyze these functions independently. CPZ was found to act mainly on the inhibition of Ca2+ uptake and not to increase Ca2+ release from the SR in a concentration of CPZ (10 microM). These results suggest that CPZ may induce the increase of intracellular Ca2+ concentration. CPZ may be safely employed to MH susceptible patients, since CPZ at a clinical concentration showed no influence on Ca2+ related functions of the skeletal muscle. PMID- 10846379 TI - [Clinical features and outcomes of mechanically ventilated patients with acute respiratory failure during treatment with immunosuppressive drugs]. AB - We examined clinical features and outcomes of mechanically ventilated patient (n = 11) retrospectively who had developed acute respiratory failure during treatments with immunosuppressive drugs. The mean APACHE II score was 22.6, and the mean lung injury score was 2.9. In eight patients chest X-ray and computed tomography showed interstitial pneumonia. Fungus and/or cytomegalovirus were isolated most often from patients with interstitial pneumonia. Observed mortality (72.7%) was significantly (P < 0.001) higher than predicted mortality (45.1%) in the APACHE II score. Patients, who were complicated with septic shock caused by fungus infection, showed poor mortality. These results suggest that the fungus and cytomegalovirus infections might be associated with poor prognosis in patients with acute respiratory failure during treatment with immunosuppressive drugs. PMID- 10846380 TI - [Midazolam-atropine lollipop for pediatric premedication]. AB - Strawberry flavored sweet lollipop containing midazolam (5 mg) and atropine (0.25 mg) was evaluated for the premedication of pediatric patients as judged by sedative and anti-anxious effects and gastric fluid volume and acidity. The subjects of this prospective, double blind, random and controlled study were 175 children aged between 6 months and 10 years. They were divided into three groups: group A patients receiving the lollipop containing only 5 mg midazolam (n = 78), group B patients receiving the lollipop containing only 0.25 mg atropine (n = 21), and group C patients receiving the lollipop containing 5 mg midazolam and 0.25 mg atropine (n = 76). The plasma midazolam concentration was measured in ten children in group A. The sedative and anti-anxious effects were scaled with four levels. The children receiving the lollipop containing midazolam (group A and C) showed a better level of the sedative and anti-anxious state. The volume of gastric fluid of group A was more than those of group B and C. The pH of group A gastric fluid was also higher than those of other groups. The correlation equation of the plasma midazolam concentration (y ng.ml-1) against time (t min) was y = 149 x e(-t/64), (r = 0.775). These results suggest that midazolam atropine lollipop is one of the favorable choices as the premedication for pediatric patients. PMID- 10846381 TI - [The compound A concentration in a low-flow anesthesia circuit using the new CO2 absorbent SPHERASORB]. AB - CO2 absorbents convert sevoflurane to fluoromethyl-2,2-difluoro-1 (trifluoromethyl) vinyl ether (compound A), whose toxicity in rats raises concern regarding the safety of sevoflurane in a low-flow system. SPHERASORB is a new CO2 absorbent which does not contain KOH. However, the reaction between SPHERASORB and sevoflurane has not been examined. We compared compound A concentration in a model circuit using SPHERASORB and the commonly used Sodasorb II. The anesthesia circuit was circulated with fresh gas flow at a rate of 1 l.min-1 containing 2% sevoflurane. Compound A concentration was measured hourly and the temperature of CO2 absorbent was monitored. The maximum concentration of compound A in the circuit was 12.2 +/- 1.4 ppm for SPHERASORB and 18.6 +/- 0.4 ppm for Sodasorb II (P < 0.05). The maximum temperature of SPHERASORB was lower than that of Sodasorb II (P < 0.05). SPHERASORB can reduce compound A formation, compared to Sodasorb II. PMID- 10846382 TI - [Anesthetic management in pediatric interventional cardiology]. AB - Interventional cardiology (n = 108) and cardiac angiography (n = 481) procedures in pediatrics performed between January 1996 and December 1998 were reviewed. Means (SD) of duration of anesthesia for interventional cardiology and cardiac angiography were 245 (130) and 152 (48) minutes, respectively (P < 0.001). Incidences of long operations requiring over 6 hours of anesthesia were 12 and 0%, respectively (P < 0.001). Incidences of hemodynamic derangements were 17 and 2.9%, and catheter-related complications were 7.4 and 0.83%, respectively (P < 0.001). Incidences of ICU admission were 7.4 and 0.62% those of emergency surgery were 4.6 and 0%, and those of blood transfusion were 4.6 and 0%, respectively (P < 0.001). Incidences of cardiopulmonary resuscitation, and cardioversion were also higher in interventional cardiology (P < 0.05), and all these emergency cases were rescued successfully. The risk of cardiac angiography is higher compared with general surgery, and the risk of interventional cardiology is higher than cardiac angiography in pediatrics. This study reconfirms that anesthesiologists should play an active role in care of pediatric patients undergoing high-risk procedures outside the operating room. PMID- 10846383 TI - [Monitoring of motor evoked potentials during insertion of iliosacral screws for reconstruction of pelvic fracture in two patients]. AB - Monitoring of myogenic motor evoked potentials (MEPs) induced by transcranial electrical stimulation has become a promising tool for intraoperative monitoring. We described 2 patients who had developed significant decrease in MEP during the insertion of iliosacral screws for reconstruction of pelvic fractures. In both patients, MEPs were successfully obtained prior to the insertion under general anesthesia and partial neuromuscular blockade (propofol, ketamine, fentanyl, and nitrous oxide in oxygen: vecuronium), but reduced in association with the insertion. In one patient, they were restored by the re-insertion of screw and no new neurological deficits were observed postoperatively. However, in another patient, the decrease was not normalized and he suffered from paresis of the lower extremities after the surgery. We consider that intraoperative changes in MEPs could precisely predict postoperative motor function. PMID- 10846384 TI - [Anesthetic management for placement of tracheobronchial stents in patients with airway stenosis]. AB - We report the anesthetic management for stents placement in patients with tracheobronchial stenosis. The subjects were 6 patients with lung cancer and one patient with tracheal invasion of esophageal cancer. Anesthesia was induced with propofol, fentanyl and vecuronium, and maintained with propofol and vecuronium. After intubation, tracheostomy was performed. The patients were kept apnic during insertion of stents. Three patients had dynamic stents inserted from tracheostomy site and one orally. Three patients had Dumon stents inserted orally, but the procedure in one patient was cancelled because her stent could not be placed at appropriate position. We recommend the anesthetic management through the tracheostomy site for the placement of Dumon tubes or dynamic stents. PMID- 10846385 TI - [Intraoperative management of heart transplantation in Japan--report of two cases]. AB - We experienced intraoperative anesthetic management of two cases of heart transplantation in Japan. Both patients were in the end stage of cardiac failure due to dilated cardiomyopathy. One patient had had implantation of left ventricular assist system, and another patient had had implantation of automated cardioveter defibrillator. Transesophageal echocardiography was useful for the monitoring of cardiac function during the operation. Anti-arrythmic therapy including heart pacing and protection of right heart failure are important for the circulatory management of heart transplantation. The anesthesiologist is needed not only for the management of respiration and circulation but also for the prevention of infection and control of the time schedule. PMID- 10846386 TI - [Utility of Ringer's acetate solution as an intraoperative fluid during cardiovascular surgery with cardiopulmonary bypass]. AB - This study aimed to clarify the difference in the effects of Ringer's acetate (AR) and Ringer's lactate (LR) administration during cardiovascular surgery with cardiopulmonary bypass. We evaluated their effects on intra and postoperative metabolism, liver functions, blood gas and hemodynamic states. Twenty patients were divided into two groups; AR group (n = 10) and LR group (n = 10). Intraoperative serum D-lactate levels in LR group were significantly higher than those in AR group from the beginning of the operation to awakening. Serum acetate levels showed no increase in both groups. The arterial ketone body ratio (AKBR) in AR group was higher than that in LR group, but the difference was not significant. Serum glutamic pyruvic transaminase (GPT) and alkaline phosphatase (ALP) levels in LR group were significantly higher than those in AR group from the induction of the anesthesia. It has been reported that acetate has a greater vasodialatory effect than lactate. However, our findings indicate no significant difference in hemodynamics between the two groups. These results suggest that AR may be more useful than LR during cardiovascular surgery with cardiopulmonary bypass. PMID- 10846387 TI - [Intraoperative hemodilutional autotransfusion using a closed circuit for patients of Jehovah's Witness]. AB - We conducted hemodilutional autotransfusion using a closed circuit combined with a cell washing reinfusing system (Cell Saver) for two surgical patients of Jehovah's Witness. One was a 12 yr-old boy for extirpation of the teratoma in the anterior mediastinum and another was a 44 yr-old woman for left total hip replacement. The patients and their relatives had consented to the use of blood substitutes, hemodilutional autotransfusion using a closed circuit and Cell Saver. We devised a closed circuit system for hemodilutional autotransfusion combined with Cell Saver, in which two pumps for blood transfusion were used; one was for drawing blood from the femoral or the internal jugular vein and the other for returning blood to the peripheral vein. Blood volume in a bag interposed in the closed circuit was easily controlled by adjusting the speed of each pump. Blood collected from the surgical field by Cell Saver was also led to the bag. Acid citrate dextrose solution was infused into the closed circuit from the site close to the blood drawing. Both of our surgical patients were safely managed without homologus blood transfusion, although there remained some problems concerning the use of anticoagulants. PMID- 10846388 TI - [Changes of latency and amplitude of short latency somatosensory evoked potentials during slow induction of anesthesia with sevoflurane]. AB - In the present study, changes of latency and amplitude of short latency somatosensory evoked potentials (SSEPs) were evaluated continuously during slow induction of anesthesia from sevoflurane awake to deep levels of anesthesia in eight scheduled surgical patients. Not consistent with other previous investigations, the latency of N20 was significantly shortened with sevoflurane after 25 minutes from the beginning of inhalation compared with the awake control levels. No increase of latency was observed. The amplitudes of N20 were decreased with sevoflurane anesthesia in relation to duration of anesthesia. These results suggest that sevoflurane alone might have no marked pharmacological properties to change the latency of SSEPs even in the deep level of anesthesia. PMID- 10846389 TI - [Relationship between changes in estimated and measured blood concentrations of propofol and serum albumin concentration during propofol anesthesia: effects of intraoperative bleeding (a case report)]. AB - We investigated the effect of intraoperative bleeding on the changes in estimated and measured blood concentrations of propofol, and the relationship between anesthetic effect of propofol and serum albumin concentration in a patient undergoing prostatectomy during propofol/epidural anesthesia. The rate of propofol infusion was titrated to keep the BIS value about 50. The estimated blood concentration of propofol calculated by ConGrase, a program for on-line simulation of blood propofol concentration, was monitored during anesthesia. Blood samples were obtained at nine points during surgery and the blood concentrations of propofol and albumin were measured postoperatively. When the amount of bleeding reached about 2000 mg, the difference between estimated and measured blood concentrations of propofol did not increase. As the serum albumin concentration decreased, the concentration of propofol to keep bispectral index about 50 decreased. This study suggests that the change in serum albumin concentration following intraoperative bleeding is an important factor in the anesthetic effect of propofol. PMID- 10846391 TI - [Anesthetic management of a patient with a huge ovarian tumor]. AB - We managed a patient with a huge ovarian tumor (15 kg). The patient was a 50-year old woman and could not take the supine position because of the tumor causing respiratory embarrassment. An epidural catheter was inserted 3 cm cephalad via the Th 11-12 interspace in the right lateral position. Three milliliters of 1% mepivacaine was injected epidurally for test dose and produced hypesthesia of Th 9-12 five min after the injection. Additional 3 ml of 1% mepivacaine was injected epidurally, which widened the hypesthesia to Th 5-L 2. Intra-arterial pressure was monitored continuously. Under epidural anesthesia without sedation, 11,000 ml of fluid was suctioned slowly from the cyst in 20 min, during which time remarkable hemodynamic derangement did not occur. The patient was turned into the supine position and the trachea was intubated. Laparotomy was performed under general anesthesia. During the surgery, respiratory and hemodynamic conditions were stable. On the following day, chest radiography demonstrated an abnormal shadow in the lower lobe of the right lung. It disappeared the next day without any treatment. Anesthetic management of patients with huge abdominal tumor is also discussed. PMID- 10846390 TI - [Coronary artery spasm during vascular surgery]. AB - A 56-year-old male with arteriosclerosis obliterans was scheduled for aorto biiliac artery bypass graft surgery. He had episodes of chest pain lasting for several minutes. Preoperative electrocardiogram (ECG) and myocardiac scintigram showed no abnormal findings. Anesthesia was induced with thiopental 225 mg, fentanyl 100 micrograms and tracheal intubation was facilitated with vecuronium 10 mg. Anesthesia was maintained with O2 2 l.min-1, N2O 4 l.min-1, sevoflurane 0.5-1% and epidural injection of 1% mepivacaine 6-10 ml.hr-1. Nitroglycerin was infused at a rate of 0.125 microgram.kg-1.min-1. During surgery, a transient elevation of the ST segment occurred three times in lead II of the ECG. For the first episode with decrease in blood pressure at declamping the external iliac artery, administration of phenyrephrine 100 micrograms improved ST segment elevation in ECG. For the second event with a slight decrease in blood pressure, an increased dose of nitroglycerin decreased ST segment elevation. The third incident with increased blood pressure and heart rate was alleviated by decreasing the dose of dopamine. Postoperative ECG and serum creatine kinase level were within normal limits. These three episodes of ST segment elevation might be due to coronary spasms induced by decreased blood pressure or increased blood pressure and heart rate. PMID- 10846392 TI - [Dyspnea attack due to hysteria after general anesthesia]. AB - A 30-year-old female was scheduled for an expander insertion of the breast under local anesthesia. Thirty minutes after infiltration anesthesia with lidocaine and bupivacaine mixture, she suffered from dyspnea. She was intubated and transferred to our hospital. As her vital signs were stable and consciousness was clear, she was extubated in the emergency room. However, she was reintubated at night and ventilated mechanically for two days. Three months later, breast expander insertion was performed under general anesthesia. After extubation, dyspnea attack occurred and midazolam was injected. Seven months later, the reconstruction of TRAM flap was performed under general anesthesia and continuous subcutaneous injection of morphine was used for the postoperative analgesia. After extubation, she was sedated deeply and dyspnea attack did not occur. A month later, she was scheduled for the debridment and the resuture. Then, dyspnea attack occurred in the ward at night. The apnea monitor was attached to her in recovery room after extubation following the operation of debridment and resuture. Dyspnea attack appeared and was diminished with midazolam injection. We diagnosed her as hysteria with CMI and MMPI psychologic tests. PMID- 10846393 TI - [An automated electronic anesthesia record using a hospital LAN (local area network)]. AB - We have developed an automated electronic anesthesia record system using a hospital LAN. As the number of monitors we can use in the operating room is increasing, it is impossible to record all physiologic parameters in a handwritten anesthesia record. Physiologic parameters are recorded every 10 seconds from the anesthesia monitor. An operation ordering system by a hospital LAN has been completed and the patient's data are stored in a host computer, and we can use its data for the automated electronic anesthesia record preoperatively. The advantages of the automated electronic anesthesia record are continuous high quality, more data collection than the handwritten anesthesia record, and the electronic database. During a critical period, the anesthesiologist is too busy to plot physiologic parameters but the automated electronic anesthesia record is reliable and accurate. Disadvantage of the automated electronic anesthesia record is some practice required to input clinical events such as drug administration. The handwritten anesthesia record is easy to use and economical. Ergonomic problems still remain to be solved for wider acceptance of the automated electronic anesthesia record in clinical practice. At the end of the operation, intraoperative data are sent to a host computer and the anesthesia record is printed. We can use this database for clinical research and retrospective case reviews. The implementation of the automated electronic anesthesia record in anesthesia practice will improve quality of patient care. PMID- 10846394 TI - [Sitting position in the neurosurgery: the results of a questionnaire sent to neurosurgeons of medical colleges]. AB - We analyzed questionnaire on the sitting position among neurosurgeons. We sent questionnaire to 80 medical colleges in Japan and obtained response from 61 universities (76.3%). The sitting position surgery was performed in 7 institutions (11.5%) and was not in other 54 (88.5%). The first reason for not adopting the sitting position was "other position was good enough to perform the surgery" (44 institutions, 81%). This reason exceeded "objection from the anesthesiologist" (17 institutions, 31%). Twenty institutions (33%) adopted or wished to perform the surgery in sitting position, indicating that neurosurgeons still are interest in performing the sitting position surgery. But 6 institutions adopting the sitting position predicted that the sitting position would diminish in the future, and the sitting position surgery would decrease in number in Japan. PMID- 10846395 TI - [Surfactant proteins A and D as biomarkers of disease activity in diffuse interstitial pneumonia]. AB - We evaluated the clinical significance of surfactant proteins A (SP-A) and D (SP D) as useful markers of disease activity in patients with diffuse interstitial pneumonia. Serum concentrations of SP-A and SP-D were measured by the sandwich ELISA method. The serum levels of SP-A and SP-D in patients with diffuse interstitial pneumonia (IIP, CVD-IP, HP, Ra-IP) were significantly higher than the levels in healthy controls, and showed high positive rates. IIP patients characterized by a predominantly ground-glass opacity (GGO) pattern on high resolution computed tomograms had significantly higher concentrations of serum SP A. Elevated SP-D levels reflected the extent not only of GGO but also of parenchymal collapse opacity (PCO). It is likely that the mechanisms behind the elevation of SP-A and SP-D do not correlate with pathologic changes in IIP. Serum SP-A and SP-D levels obtained at the time of initial evaluation from 9 patients who died after less than 3 years of follow-up were significantly higher than in patients with survival rates of more than 3 years. Serum SP-A and SP-D may be useful biomarkers of disease activity in patients with IIP. PMID- 10846396 TI - [Effectiveness of domiciliary noninvasive positive pressure ventilation in improving blood gas levels and the performance of daily activities. NIPPV Study Group]. AB - Noninvasive positive pressure ventilation (NIPPV) is widely used in the domiciliary treatment of chronic ventilatory failure. We conducted a multicenter, prospective study of the effects of NIPPV on blood gas levels, subjective symptoms, and the performance of daily activities. Fifty-one hospitalized patients with chronic ventilatory failure were enrolled in the study. Using NIPPV, 44 patients were able to live at home for more than 1 month, and 43 for more than 3 months. PaO2 and PaCO2 were 69.1 +/- 13.1 Torr (n = 25) and 70.8 +/- 11.0 Torr (n = 27) before NIPPV and 77.9 +/- 12.5 Torr (n = 26) and 61.0 +/- 11.2 Torr (n = 27) after 1 month of domiciliary treatment, respectively. Blood gas values were significantly improved after NIPPV. Patients experienced alleviation of all subjective symptoms. Dyspnea when wearing clothes, walking on flat levels, or climbing stairs also subsided, and was significantly reduced during daily activities other than eating among those patients who needed any help in performing their activities prior to the initiation of NIPPV. We concluded that NIPPV is an effective and convenient means of domiciliary ventilation that can help patients achieve greater independence in daily life. NIPPV can be expected to play a central role in home respiratory care for patients with hypercapnic respiratory failure. PMID- 10846397 TI - [Effects of inhaled corticosteroid in elderly patients with chronic obstructive pulmonary disease]. AB - We examined the efficacy of inhaled corticosteroid (beclomethazone dipropionate: BDP) in elderly patients with chronic obstructive pulmonary disease (COPD). Eighty-six patients with COPD were divided into 3 groups: COPD treated without BDP (group 1, n = 26), COPD treated with BDP (group 2, n = 25), and BDP-treated COPD with asthmatic symptoms (group 3, n = 35). Pulmonary function test findings, symptoms, and prognosis for the 3 groups were retrospectively compared. No significant differences in yearly decline of FEV1.0 were observed. Of the patients treated with inhaled corticosteroid (groups 2 and 3), 30% exhibited improved FEV1.0. Of these patients, monthly decline of FEV1.0 correlated negatively with bronchial reversibility in FEV1.0 before and after inhalation of salbutamol (delta FEV1.0) (r = -0.28, p = 0.03). Cough and sputum scores were significantly improved in groups 2 and 3 (p < 0.01). Fewer admissions and episodes of acute exacerbation were noted in groups 2 and 3 than in group 1. From these results, we concluded that inhaled corticosteroid was effective in elderly COPD patients with bronchial reversibility and airway symptoms. PMID- 10846398 TI - [Clonal rearrangement of intratumoral T-cell receptor beta-chain gene in two patients with thymoma accompanied by pure red cell aplasia]. AB - We encountered two cases of thymoma accompanied by pure red cell aplasia and demonstrating clonal rearrangement of the T-cell receptor beta-chain gene (TCR beta) in lymphocytes. Patient 1 was a 55-year-old man and Patient 2 was a 43-year old woman. Both had severe anemia and mediastinal tumors. Bone marrow aspiration was performed and pure red cell aplasia diagnosed. Thymoma was the presumptive diagnosis for the mediastinal tumors, and extended thymectomy was performed. The post-operative diagnosis was invasive thymoma (spindle-cell type) in Patient 1 and non-invasive thymoma (mixed lympho-epithelial type) in Patient 2. The cell compositions (%) obtained with T-cell surface marker analysis were as follows: [table: see text] Southern blot analysis disclosed clonal rearrangement of TCR beta genes in thymoma thymocytes from both patients. PMID- 10846399 TI - [Pulmonary actinomycosis presenting as a mass lesion on chest X-ray film]. AB - A 61-year-old man presented with fever, productive cough, and occasional blood streaked sputum. Chest X-ray films disclosed a poorly defined mass in the right middle lung field. A transbronchial lung biopsy specimen showed epithelial changes indicative of lung cancer, and a right lower lobectomy was performed. Because the resected specimen contained a cavity filled with colonies of actinomycetes, pulmonary actinomycosis was diagnosed. The cavity was surrounded by inflammatory infiltrations and fibrosis with occasional atypical epithelial changes suggestive of benign hyperplasia. Although pulmonary actinomycosis is uncommon today, it deserves attention because of the potential difficulty in differentially diagnosing it from lung cancer. PMID- 10846400 TI - [Sjogren's syndrome with malignant lymphoma, interstitial pneumonia, and pulmonary hypertension]. AB - A 62-year-old woman was admitted to our hospital for further examination of cough and dyspnea on exertion. She had suffered xerostomia and xerophthalmia for 7 years. Physical examinations showed purpura on the lower extremities, Raynaud's phenomenon, and swelling of the parotid glands. Laboratory data disclosed thrombocytopenia, hypergammaglobulinemia, and high titers of anti-nuclear antibody, anti SS-A, and anti SS-B antibodies. Sicca symptoms, a positive Schirmer's tear test, and laboratory findings together yielded a diagnosis of Sjogren's syndrome. Chest X-P and computed tomographic films demonstrated diffuse reticulonodular shadows and multiple nodules in the left lower lung. Echocardiography and right cardiac catheterization revealed enlargement of the right ventricle, tricuspid regurgitation, and elevated pulmonary arterial pressure. Histologic findings from an open-lung biopsy specimen revealed accumulation of inflammatory cells and fibrosis around broncho-vascular bundles, and diffuse large cell lymphoma. Despite a favorable response to chemotherapy, the patient died of right ventricular failure. PMID- 10846401 TI - [Rheumatoid arthritis-associated bronchiolitis successfully treated with erythromycin]. AB - A 52-year-old man with a 4-year history of rheumatoid arthritis, and who had an episode of suspected BOOP in early 1994, was admitted to our hospital because of cough and fever. A chest X-ray film on admission showed small patchy infiltrates, and a computed tomographic (CT) scan showed centrilobular nodules and patchy infiltrates with thickened broncho-vascular bundles in both lungs. Transbronchial and thoracoscopic lung biopsies disclosed the coexistence of interstitial pneumonia with BOOP pattern, follicular bronchiolitis, and diffuse panbronchiolitis-like purulent and obliterative bronchiolitis. Due to findings of chronic sinusitis, the patient was treated with erythromycin for 8 weeks, and the abnormal CT shadows regressed. This was an interesting case of various pulmonary lesions associated with rheumatoid arthritis, and successfully treated with erythromycin. PMID- 10846402 TI - [Myeloperoxidase-antineutrophil cytoplasmic antibody positive alveolar hemorrhage during propylthiouracil therapy for hyperthyroidism]. AB - A 62-year-old woman had been treated with propylthiouracil(PTU) for hyperthyroidism. Because bloody sputum, dyspnea, and severe hypoxemia developed, the patient was admitted to our hospital. Chest X-ray and chest computed tomographic (CT) films disclosed diffuse infiltrative shadows in both lung fields. Bronchoalveolar lavage revealed abundant hemosiderin-laden macrophages. Alveolar hemorrhage associated with myeloperoxidase-antineutrophil cytoplasmic antibody (MPO-ANCA) positive vasculitis syndrome was diagnosed because of the high serum level of MPO-ANCA. After the initiation of steroid therapy and termination of PTU, the infiltrative shadows in both lung fields disappeared, PaO2 improved, and MPO-ANCA decreased. There have been some reports of MPO-ANCA positive vasculitis syndrome developing during PTU therapy, but most were concerned with renal disease. We concluded that PTU and similar agents should be given consideration as one of the possible causes of MPO-ANCA-induced alveolar hemorrhage. PMID- 10846403 TI - [Pulmonary involvement in acute febrile neutrophilic dermatosis (Sweet's syndrome)]. AB - We encountered a 55-year-old man with pulmonary involvement in acute febrile neutrophilic dermatosis (Sweet's syndrome). He had been treated with steroids for Sweet's syndrome for 2 years, and on September 17, 1998 presented with a cough and a fever of 38.9 degrees C. Physical examination revealed fine crackles at the bases of both lungs. Chest radiography and computed tomography demonstrated reticular and nodular infiltrates in both lungs. Treatment with a variety of antibiotic agents and an antifungal agent was not effective. Sputum culture was sterile and bronchial washings were negative for infectious pathogens. Transbronchial biopsy revealed a mild chronic interstitial infiltrate and an inflammatory exudate in bronchiolo-alveolar tissue. The pulmonary lesions and cutaneous lesions were resolved by intradermal injections of triamcinolone acetonide in addition to oral prednisolone. Although the apparent neutrophilic infiltrates cited by earlier reports were not observed in transbronchial biopsy specimens, the clinical course in this case suggested that our patient had Sweet's syndrome with pulmonary involvement. PMID- 10846404 TI - [Two cases of multiple primary cancer involving the lung with old pulmonary tuberculosis]. AB - We reported 2 relatively rare cases of multiple primary cancer including lung cancer accompanied by old pulmonary tuberculosis. Patient 1 was a 62-year-old man admitted to our hospital for further evaluation of an infiltrative shadow on chest X-ray films, and a cervical tumor noted 10 years earlier and thought to be thyroid cancer. A Transbronchial lung biopsy (TBLB) specimen disclosed poorly differentiated squamous cell carcinoma. A right upper lobectomy and thyroidectomy were performed. Histopathologic findings showed a neoplastic lesion adjacent to caseous necrosis with formation of granuloma consistent with tuberculosis. Also, the cervical tumor was considered to be a metastatic lymph node from thyroid papillary carcinoma. Patient 2 was a 73-year-old man with a 14-year history of treatment for transitional cell carcinoma of urinary bladder, who had been admitted to our hospital for further evaluation because of a nodular shadow observed on chest X-ray films. TBLB specimens disclosed adenocarcinoma. A right upper lobectomy was performed. Histopathologic findings revealed a neoplastic tumorlet in the same lobe. No detectable increases in serum TNF-alpha, IL-1 beta or IFN-gamma were observed in either patient. Phytohemagglutinin- and concanavalin-A-stimulated lymphocyte proliferation decreased in Patient 1. These findings suggested that the immunocompromised status of patients with cancer in addition to old pulmonary tuberculosis may contribute to the development of lung cancer. PMID- 10846405 TI - [Microscopic polyangitis with pleuritis as the only pulmonary complication]. AB - A 75-year-old woman was admitted to our hospital because of high fever and appetite loss. A chest roentgenogram and computed tomographic scans revealed pleural effusion without obvious infitrative or interstitial shadows in both lung fields. Laboratory data showed microhematuria, proteinuria, and telescoped sediment with a moderate increase in C-reactive protein, suggestive of acute glomerulonephritis. Because infectious pleuritis, was initially suspected, the patient was treated with antibiotics. However, her general condition deteriorated, and the right pleural effusion increased. Levels of myeloperoxidase specific anti-neutrophil cytoplasmic antibody (MPO-ANCA) in serum and pleural effusion were markedly elevated, yielding a conclusive diagnosis of MPO-ANCA related vasculitis, especially microscopic polyangitis (MPA). The Patient was immediately treated treated with prednisolone, cyclophosphamide, and plasma exchange. Several weeks later, her general condition dramatically improved, and the level of MPO-ANCA in serum markedly decreased. In addition, the pleural effusion completely disappeared. Unfortunately, the patient eventually died of opportunistic infections (MRSA-pneumonia and Aspergillus-pneumonia) 6 months after admission. This was a unique case of MPA associated with pleuritis without interstitial pneumonia or alveolar hemorrhage. PMID- 10846406 TI - [An adult case of cervico-mediastinal tuberculous lymphadenitis]. AB - We reported a case of cervico-mediastinal tuberculous lymphadenitis followed by the development of pulmonary lesions and recurrent laryngeal nerve palsy. A 48 year-old man was admitted to our hospital due to fever, dry cough, and loss of body weight. He had no medical history of tuberculosis or other significant diseases. On admission, a right cervical tumor was palpable and chest X-ray films revealed widened superior mediastinum. Chest computed tomography showed multiple swollen mediastinal lymph nodes, including multiple low-density areas and contrast medium-enhanced septa and margins. Gallium-67 scintigraphy demonstrated abnormal uptake in the right cervix and mediastinum. A PPD skin test was strongly positive and ribosomal RNA of tubercle bacilli was detected in aspirated gastric juice. Although anti-tuberculous chemotherapy was initiated, fever and cough persisted, and hoarseness due to left recurrent laryngeal nerve palsy developed. One month later, chest X-ray films showed abnormal infiltration in the left upper lung field. The patient was sero-negative for HIV. To confirm the diagnosis, right supraclavicular lymph node biopsy was performed. Microscopic examination of the biopsy specimen revealed acid-fast bacilli and granuloma with central caseous necrosis surrounded by Langhans' giant cells and epithelioid cells. After 9 months of extended anti-tuberculous chemotherapy, the cervical and mediastinal masses receded and the abnormal chest X-ray shadows disappeared. Because the incidence of tuberculosis in Japan is gradually increasing among young people as well as the elderly, the differential diagnosis of this disease will become more necessary. PMID- 10846407 TI - [Neurally mediated syncope in association with small cell lung carcinoma]. AB - A 66-year-old woman was admitted to our hospital because of frequent syncopal episodes and for treatment of small cell lung carcinoma. Neurally mediated syncope was diagnosed by the head-up tilt test, which evoked early severe hypotension (after 12 min at the 80-degree tilt position). Treatment of carcinoma by chemotherapy and radiotherapy promptly eliminated the syncopal episodes. This was an unusual case of neurally mediated syncope associated with small cell lung carcinoma. PMID- 10846408 TI - [Small adenocarcinoma of the lung under long-term observation]. AB - A 68-year-old man presented with symptoms of common cold at our hospital on March 8, 1995. Ground glass opacity (GGO) was detected in the right S 4 a by high resolution computed tomography (HRCT) during a routine examination. Except for mild traction of the nearest pleura, the lesion itself demonstrated little change on HRCT images for 2 years. HRCT images disclosed slight enlargement of the lesion over a long-term period of follow-up observation. Exactly 25 months after the first examination, HRCT scans demonstrated an area of centralized dense concentration. Right middle lobectomy was performed 30 months after the lesion was first detected. Pathologic findings from the resected specimen revealed Noguchi's small adenocarcinoma of the lung (type B classification: localized bronchioloalveolar carcinoma with collapse of alveolar structure). The lesion was scanned 8 times with HRCT during the follow-up period. The change to GGO with centralized density and spicula was observed in detail, and thought to be evidence of a progression from type A to type B. We concluded that a set of fixed settings should be utilized for repeated HRCT examinations of small peripheral lesions exhibiting GGO. PMID- 10846409 TI - [Clinical problems of NSAID-associated ulcer]. PMID- 10846410 TI - [Exocrine pancreatic zymogen activation]. PMID- 10846411 TI - [Clinical evaluation in oral lichen planus with chronic hepatitis C: the role of interferon treatment]. AB - Hepatitis C virus (HCV) infection induces a variety of extrahepatic manifestations such as oral lichen planus (OLP). To clarify the role of HCV in the development of OLP, we investigated the occurrence of OLP in patients with chronic hepatitis C treated with interferon (IFN). Of 275 patients with chronic hepatitis C, 6 developed OLP during the IFN treatment. However, OLP developed in none of 230 patients with chronic hepatitis C who did not undergo the IFN therapy. The IFN treatment in chronic hepatitis C patients developed OLP significantly, as compared with the non-treated group (p < 0.05). 4 of 6 patients who developed OLP during the IFN treatment had a complete response with normalization of ALT levels and undetectable HCV RNA after the treatment. There were no significant correlations between the effect of the IFN treatment and outcome of OLP. Furthermore, 3 of the 6 patients developed OLP, when serum HCV RNA became negative. These results suggest that direct viral factors may not be important in the pathogenesis of OLP in patients with chronic hepatitis C. Immunological changes caused by IFN may play a role in the development of OLP associated with HCV infection. PMID- 10846412 TI - [Two cases of ocular damage associated with proton pump inhibitors]. PMID- 10846413 TI - [A case of preoperatively diagnosed signet ring cell carcinoma of the appendix vermiformis]. PMID- 10846414 TI - [Two cases of transcatheter therapy to hepatocellular carcinoma supplied by the right internal mammary artery]. PMID- 10846415 TI - [A case of infected liver cyst perforating into the omental sac]. PMID- 10846416 TI - [A case of intrabile duct hepatocellular carcinoma presenting a stone-like appearance in the common bile duct on an endoscopic retrograde cholangiogram]. PMID- 10846417 TI - [A case of a giant solid cystic tumor of the pancreas with lymph nodes metastases]. PMID- 10846418 TI - [A case of pancreatic pseudocyst intruded into the left lobe of the liver]. PMID- 10846419 TI - [A case of pancreatic pseudocyst penetrating into the spleen associated with primary hyperparathyroidism]. PMID- 10846420 TI - [A case of anisakiasis of the rectum treated under colonoscopy]. PMID- 10846421 TI - [Hemostasis in liver diseases]. AB - Liver plays central role in the synthesis and metabolism of the pro- and anticoagulant enzymes of blood coagulation. Acute or chronic liver failure frequently result in different bleeding phenomena. Thrombocytopenia due to hypersplenism and lack of haemopoietic factors seems to be common, but in spite of thrombocytopenia and more or less platelet malfunction thrombocytopenic bleeding is usually less prominent feature. Bleeding esophageal varices, clotting abnormalities with peritonejugular shunts may pose many difficulties in clinical practice. Transjugular intrahepatic shunt (TIPS) has been a major step forward treating refractory esophageal bleeding or ascites, however to keep the stent patent seems to be still unresolved. Along with the bleeding tendency or symptoms concommittant venous thromboembolic events are may not be considered as rare events in cirrhosis. Special problems are also coupled with ascites, which contains an almost full inventory of coagulation proteins, also with the medical and interventional therapy of Budd-Chiari syndrome, and other veno-occlusive conditions with or without transplantation. PMID- 10846422 TI - [Use of positive suggestions in medical practice: experiences in the intensive care unit]. AB - The stressful states (e.g. the state of illness, sickness, acute or chronic pain, comatose and perioperative states) can be considered as psychological states in which we are more susceptible to all suggestions, including the deliberate or involuntary, direct or implicit ones. Under these conditions whether conscious of unconscious, peoples' thinking processes change, become literal, direct and somewhat paranoid, overly sensitive for not only to the direct, but to the implicit, inner meanings of communicative messages, as well. Research evidence shows that even the comatose people may maintain contact with the environment, therefore the conversation around and other communicative effects may serve as a special suggestion. These messages can be (unconsciously) interpreted by the patients following different logic than it would have been processed in the waking state. Treating people who are in these special stressful states one should carefully analyze all of the meaning layers of his/her communication, to make the helpful, positive suggestions more effective, and avoid the negative, destructive ones. The possible negative suggestions are those words, labels, and actions, that may be proper from practical, economical or organisational point of view, but we do not take their implicit suggestive meanings into considerations. The paper gives a detailed overview about the role of suggestions used during the management of a patient in intensive care situations, and some guidelines are outlined to make our (possible) negative suggestions to be helpful and positive. PMID- 10846423 TI - [Experiences with josamycin therapy in dermatologic diseases]. AB - Josamycin belongs to the new generation of macrolide antibiotics. Josamycin treatment was applied to various dermatological diseases (acne, pyoderma) and sexually transmitted bacteriological infections. During the examinations in ambulance treatment, 26 patients out of a total of 55 became completely healthy, 27 got better and in case of 2 persons, their conditions did not change. 4 of the sick people reported of side effects. According to the author, josamycin could be a favourable choice in certain dermatological pathographies and sexually transmitted infections because of its effectiveness and good tolerance. PMID- 10846425 TI - [Rhabdoid meningioma: a potentially aggressive new variant]. AB - Rhabdoid meningioma is a recently recognized clinicopathologic entity characterized histologically by cytoplasmic aggregates of intermediate filaments, and clinically by the propensity of such tumors to pursue an aggressive course. The authors report on clinical, radiologic and pathologic findings in three cases of rhabdoid meningioma identified in a retrospective surgical series of 204 meningothelial tumors. Patients included two females, aged 39 and 55 years, and a 54-year-old male. In the first two cases the tumors were located on the right and left lesser sphenoid wing, respectively; in the third case, the right cerebellopontine angle was affected. All three neoplasms evolved on a background on transitional meningioma and were conspicuous for dis-cohesive tumor cells and suppression of syncytical architecture. Immunohistochemistry and ultrastructural examination confirmed the meningothelial origin of inclusion-bearing rhabdoid cells. Although none of the tumors showed evidence of histologic anaplasia and Ki 67 labeling indices remained inferior to 2%, infiltrative growth into adjacent brain was noted in all three cases. On follow-up ranging from 8 months to 6 years, the patients remained either disease-free or alive with nonprogressive residual tumor. On account of their clinical behavior, well-differentiated rhabdoid meningiomas will be accommodated in the category of atypical meningiomas (WHO grade II). Their pathogenesis is likely to involve disrupted cytoskeletal integration of cell motility and proliferation, of which the rhabdoid phenotype may possibly represent a morphologic correlate. PMID- 10846426 TI - [Comments about the minimally invasive lumbar sympathectomy]. PMID- 10846424 TI - [Clinical experience with rhenium-188 HEDP therapy for metastatic bone pain]. AB - Rhenium-188 hydroxyethylidene diphosphonate (Re-188 HEDP) is a new radiopharmaceutical for treatment of metastatic bone pain. Re-188 is a generator produced radionuclide emitting high energy beta and gamma rays and having a relative short physical half-life makes it of especially interesting for therapeutic purpose. Seven patients (pts) with multiple painful bone metastases were treated with Re-188 HEDP. Five pts with prostate cancer and 2 pts with breast cancer received a fixed activity of 3000 MBq of Re-188 HEDP intravenously in two steps. Complete blood counts were determined, blood chemistry examinations and urine-analysis were performed before and 1, 2, 3, 4, 6, 8, 12 weeks following the treatment. A visual analogue score, a verbal rating scale, the Spitzer index and the Karnofsky score were used to assess pain and performance status. Three hours after Re-188 HEDP administration at 1 m from the anterior mid-trunk of the pts gamma and at the patient body surface beta-radiation dose measurements were made, together with urine radioactivity measurements. Three pts become pain-free, 2 pts exhibited partial pain improvement and 1 patient gave no response to the Re 188 HEDP therapy. In 1 patient due to central nervous system metastasis the modification of the pain intensity could not be evaluated. Three pts displayed a flare reaction within 1 week after the treatment. Transient decreases in platelet and white blood cell counts were observed. There were no significant changes in the liver and renal functions. Radiation dose rate values of 6.3 +/- 1.0 microSv/h for gamma, and of 183 +/- 40 s-1 for beta-radiation were found. 25-32% of the administered dose was eliminated via the urinary tract in the first three hours. The preliminary data suggests that Re-188 HEDP is an effective radiopharmaceutical in treatment for metastatic bone pain. An administered activity of 3000 MBq can bring about a pain reduction without causing any clinically significant bone marrow toxicity. PMID- 10846427 TI - [Current diagnostic methods for papillary carcinoma of the thyroid]. PMID- 10846428 TI - [Alcohol consumption and risk of stroke]. PMID- 10846429 TI - [The first complete sequence of a human chromosome]. PMID- 10846430 TI - [Evaluation of clinical competence: to describe or to reconstruct?]. PMID- 10846431 TI - [Is a renal biopsy really necessary in current clinical practice?]. PMID- 10846432 TI - [Leptin and obesity: is the use of this hormone the solution to this illness?]. PMID- 10846433 TI - [New horizons in influenza prevention]. PMID- 10846434 TI - Assessment of residual disease in acute leukemia by means of polymerase chain reaction. AB - Along a 5-year period in a single institution, specific molecular markers were prospectively looked for in consecutive patients with acute leukemia, by means of polymerase chain reaction (PCR): In patients with acute lymphoblastic leukemia (ALL), the BCR/ABL and TEL-AML1 fusion transcripts as well as clonotypic immunoglobulin gene rearrangements were investigated, whereas in patients with acute myelogenous leukemia (AML) the PML-RAR alpha, AML1-ETO and CBF beta-MYH11 fusion proteins were assessed. Specific molecular markers were identified in 15/75 patients: Four with ALL (three with clonotypic IgG rearrangements and one with BCR/ABL) and 11 with AML (nine with the PML/RAR alpha fusion protein--M3 AML , and two with the AML1/ETO fusion protein--M2 AML-). During follow-up periods ranging from 1 to 60 months, seven patients cleared the residual disease assessed by PCR (RD-PCR), whereas eight patients had either persistence of RD-PCR or a molecular relapse. For patients without or with RD-PCR, the 30-month survival (SV) was 86% and 14%, respectively, whereas median SV was > 60 and two months, also respectively (p < 0.01). Six of eight patients with detectable RD-PCR died, all of them within three months after the detection of the RD-PCR, whereas two of the patients that relapsed were rescued with treatment and entered a second molecular remission. Two of the three molecular relapses were detected without an overt morphological relapse. It is concluded that PCR is a valuable method for assessing residual disease and that early diagnosis of relapses may lead into effective salvage treatment in some instances. PMID- 10846435 TI - [Percutaneous kidney biopsy, analysis of 26 years: complication rate and risk factors; comment]. AB - Percutaneous renal biopsy is an invasive procedure that can result in major and minor complications. The objective of this study was to know the frequency and type of complications in relation with this procedure, as well as the efficacy to obtain enough material for diagnosis. METHODS: Retrospective study. We review the charts of patients to whom a percutaneous renal biopsy of native kidneys was done between January 1970 and March 1996. The following data were obtained: age, gender, clinical and histopathological diagnosis, complications associated with the procedure (minor: hematuria, local infections, hematoma; major: transfusions, severe infections, surgery, nephrectomy, arteriography, embolism and death). RESULTS: We analyzed 1,005 renal biopsies in 840 patients, mean age 37.7 +/- 13.1 years, 67% female. There were no complications in 88.8% (893 biopsies), minor complications in 8.65% (87 biopsies) and only in 2.4% of the procedures major complications. We divided the cases in two groups: percutaneous renal biopsy without complications (n = 893, 89%) and with complications (n = 112, 11%). The most frequent complications were hematuria (91 cases, 9.1%) and perirenal hematoma (29 cases, 2.7%). In these cases transfusion was required in 2.4% (26). Infectious complications were: urosepsis in 7 cases (0.7%), bacteremia, sepsis and perirenal abscesses (1 case each, 0.1%). One patient died because of multiple complications (0.1%). We observed greater risk of major complications on patients in those who biopsy was done because of acute renal failure (OR 4.03, p < 0.003). DISCUSSION: In our experience percutaneous renal biopsy is a low risk procedure. Most complications are minor and without clinical repercussion. There must be a strict selection criteria of the patients to whom percutaneous renal biopsy is going to be done because of the risk of severe complications. PMID- 10846436 TI - [Clinical competence in systemic arterial hypertension of undergraduate students at 2 schools of medicine]. AB - OBJECTIVES: A) To develop and validate a questionnaire for exploring clinical competence of medical students in Hypertensive Vascular Disease. B) To know and compare clinical competence among medical students of two medical schools, who are about to be caring for hypertensive patients. DESIGN: Cross-sectional study. METHODS: The questionnaire was developed with actual clinical cases and 150 items for exploring clinical abilities; experts, medical attendants in the Mexican Social Security Institute validated it. The questionnaire had to be answered by students from two medical universities about to graduate. RESULTS: We obtained a Kuder Richardson coefficient of 0.89. There were significant differences between students when we compared total scores, especially in three of the nine indicators explored. In the group 1, 94% of its students has clinical competence qualifying as low and very low, and; 87% of group 2 has clinical competence qualifying as intermediate or low. When we explored the items corresponding to treatment there is no significant difference and almost all the students are in clinical competence low and very low or as if they hadn't studied medicine. CONCLUSIONS: We consider that the questionnaire was valid and reliable. The clinical competence of the students included from universities that were included in this study, is deficient in resolving the clinical cases presented in the questionnaire. Therefore, it is probably that they will not give adequate clinical care to patients with hypertension, so it is important to improve the clinical competence of the future general practitioners, at least during their year of social service. PMID- 10846437 TI - [High-risk eating behavior in Mexican adolescents. Data on the student population in the Federal District]. AB - Data of the Drug and Alcohol Prevalence in Student Population of Mexico City Survey 1997 were used to achieve knowledge about the presence of risk eating behaviors, characteristic of eating disorders in the student population of Mexico city (N = 10,173). The sample was conformed by boys (47.9%) and girls (52.1%), with a mean age of 14.5 years (SD = 1.8). The identification of the differences between boys and girls, and age groups in two occurrence categories of risk eating behavior (ever and two or more times per week) were among the main objectives. An eleven items scale with three answering options about eating behavior during the last three months was used. The BMI-P distributed normally, with greater percentage of overweight, than underweight in both sexes. A greater percentage of risk eating behaviors were found among girls, excluding hard exercising and binge eating, as well as similar percentages of laxative and enema use in both sexes. The analysis held by age groups for risk eating behaviors showed significant differences in binge eating, self induced vomiting, pills and enemas use among boys, while girls showed differences in almost all of these practices, being the group of 18 and 19 years where the greater percentages were found. PMID- 10846438 TI - [Estimation of gasometric values at different altitudes above sea level in Mexico]. AB - We calculated reference values for arterial blood gases at different altitudes in Mexico assuming that sea level PaCO2 is 40 Torr, and in Mexico City (2.24 km. above the sea level) is 31.13 Torr, average of reported reference values. With the previous two points, it is possible to calculate a linear regression: PaCO2 = 40-3.96(altitude in km.). The equation is very similar to that calculated from reports in alveolar gas in North-Americans (Fitzgerald < 5 km): PACO2 = 39.3 3.11(altitude in km), and from subjects acclimatized to acute altitude exposure (< 5 km): PACO2 = 38.3-2.5 (altitude in km). It is also similar to a alinear equation that can be calculated assuming that hyperventilation in permanent habitants of moderate altitudes is inversely proportional to inspired molar concentration of O2: PaCO2 = PIO2/3.74. On the other hand, the equation is very different than that obtained from Andean natives (Hurtado): PaCO2 = 40.4 1.35(altitude in km). The proposed linear equation for Mexico gives very similar results (< 2 Torr difference) than a complex curvilinear equation by Morris et al. appropriate only up to 2.3 km. Evidence from acute exposure to altitude (acclimatized) and in North-Americans (alveolar gas) supports a reasonably accurate linear relationship up to 4 km. and also that the increase in ventilation in response to moderate altitudes in adult permanent residents is inversely proportional to molar concentration of O2. PAO2 was calculated with alveolar gas equation and resting the P(A-a)O2 we obtained PaO2. In conclusion, according to reference values in Mexico City, PaCO2 decreases about 4 Torr per km of altitude above the sea level. The decrease is similar to that reported in North-Americans and in acute exposure to altitude (acclimatized), but much less than that reported in native Peruvians. Ventilation is inversely proportional to the molar concentration of O2 at least up to an altitude where SaO2 is at or above 90%. PMID- 10846439 TI - Superoxide dismutase and Naproxen in the very late phase of carrageenan induced edema in rats. AB - BACKGROUND: Rat hind-foot carrageenan induced edema (CIE) is a widely used model to evaluate anti-inflammatory drugs. It shows two well-defined phases, however a third not fully characterized phase has been observed. Superoxide dismutase (SOD) is a free radical-scavenger with anti-inflammatory activity. In our country there is not a specifically designed instrument to evaluate edema in CIE. METHODS: Edema was induced by intraplantar injection of carraageenan. Fours groups were evaluated: placebo, two different doses of SOD, and Naproxen. Edema was evaluated by a specially designed mercury plethismograph. Intensity of the inflammatory reaction was determined during the classical early and late phases and during the third very late phase. RESULTS: All treatments, but higher dose of SOD, showed good anti-inflammatory activity throughout early, late, and very late phases. Naproxen was more effective than SOD during first and second phase, however this difference disappeared during the third phase. In terms of equimolar basis the enzyme appeared 1,800 time more potent than Naproxen. Mercury plethysmometer performance was fine. CONCLUSIONS: The anti-inflammatory activity of SOD and Naproxen is extended until the very late phase in the CIE model. In this bioassay, this long lasting activity of the enzyme should be ascribed to a mechanism supplementary to its free radical-scavenger property. SOD may be indeed an alternative treatment in inflammatory diseases. PMID- 10846440 TI - [Smoking among Mexican physician. A comparative analysis with smokers who are not physicians]. AB - OBJECTIVES: To determine prevalence, addiction knowledge and attitude on tobacco smoking in a group of smoking physicians (MF) and to compare these variables with smoking non-physicians (FNM) and non-smoking physicians (MNF) from the National Institutes of Health in Mexico (Insalud). MATERIAL AND METHODS: The results of a questionnaire among the three groups were compared. RESULTS: The prevalence of MF (22%) was significantly lower than in FNM (28%), (OR = 0.72, CI = 0.61-0.85). No significant differences regarding addiction and attitudes were found between them. The MNF had better knowledge and attitudes and agreed that their Institute should be a non-smoking area. CONCLUSION: Prevalence of smoking is lower among physicians than among FNM and the similarities between them suggest that addiction can provoke them and that a program for tobacco control is required. PMID- 10846441 TI - [Emergency contraception: a simple, safe, effective and economical method for preventing undesired pregnancy]. AB - In the following article, the most recent knowledge on emergency contraception (EC) is reviewed. EC is defined as those contraceptive methods that may be used to prevent an unwanted pregnancy up to 3 days after unprotected intercourse, contraceptive failure or rape. In case of non-hormonal methods (IUD), the time window for pregnancy prevention goes up to 5 days after intercourse. The different regimens now available, hormonal and non-hormonal methods, indications, contraceptive effectiveness, side effects and safety profile, possible mechanisms of action and counseling strategies will be reviewed. The potential benefits on reproductive health of wide-spread knowledge and easy, non-restrictive access to this methodology are emphasized. An extensive list of recent references is enclosed. PMID- 10846442 TI - [Limitations of the diagnostic criteria for type 2 diabetes and glucose intolerance]. AB - Conflicting results have been published during the past three decades regarding the cut points for the diagnosis of diabetes and glucose intolerance. Two major consensus changed the diagnostic values; the last revision occurred in 1997. After the publication of the new criteria, the controversy grew. New evidence was published demonstrating that the new cut points were inappropriate. The purpose of this review paper is to summarize the currently available information useful to evaluate the sensibility and specificity of the diagnostic criteria and to discuss the difficulties to define properly the cut-points for the diagnosis of diabetes and glucose intolerance. PMID- 10846443 TI - [Portal hypertension. Current status of multidisciplinary management]. PMID- 10846444 TI - [Importance of biotin metabolism]. AB - Biotin is a water soluble enzyme cofactor that belongs to the vitamin B complex. In humans, biotin is involved in important metabolic pathways such as gluconeogenesis, fatty acid synthesis, and amino acid catabolism by acting a as prosthetic group for pyruvate carboxylase, propionyl-CoA carboxylase, beta methylcrotinyl-CoA carboxylase, and acetyl-CoA carboxylase. Carboxylases are synthesized as apo-carboxylases without biotin and the active form is produced by their covalent binding of biotin to the epsilon-amino group of a lysine residue of the apocarboxylases. This reaction is catalyzed by the holo-carboxylase synthetase. The last step in the degradation of carboxylases, the cleavage of the biotinyl moiety from the epsilon-amino group lysine residues, is catalyzed by biotinidase and results in the release of free biotin, which can be recycled. Biotin regulates the catabolic enzyme propionyl-CoA carboxylase at the posttranscriptional level whereas the holo-carboxylase synthetase is regulated at the transcriptional level. Aside from its role in the regulation of gene expression of carboxylases, biotin has been implicated in the induction of the receptor for the asialoglycoprotein, glycolytic enzymes and of egg yolk biotin binding proteins. Biotin deficiency in humans is extremely rare and is generally associated with prolonged parenteral nutrition, the consumption of large quantities of avidin, usually in the form of raw eggs, severe malnutrition and, inherited metabolic disorders. In humans, there are autosomal recessive disorders of biotin metabolism that result from the disruption of the activity of biotinidase or holo-carboxylase synthetase. PMID- 10846445 TI - [Surgical management of complicated portal hypertension]. PMID- 10846446 TI - [Medical technology and health. A lot of crunching but few nuts?]. AB - The present work pretend to describe the development achieved in the field of health technology assessment and to propose a set of criteria to evaluate them, with the intention that it will lead to the development of health programs and policy with a tendency to maximize effectiveness, efficiency and quality, within a frame of increasing needs and scarce technical and economic resources for health care. For this work, a comprehensive review was carried out about the background of health technology adoption in our country, and the context by which to evaluate the accessibility and utilization of the health technology. In the section for technology assessment, it is propose a method to evaluate based upon the natural history of disease, that is, the traditional form of health assessment is restricted to a cross section in time (vertical evaluation), and in this study the authors propose a model with a horizontal approach, that should offer as its main virtues the development of guidelines with regards to prevention, diagnosis and treatment, in addition to cost containment, in support to a more effective and higher quality medical practice. PMID- 10846447 TI - [Effect of chloroquine treatment of porphyria cutanea tarda on the associated liver disease]. PMID- 10846448 TI - [Function of TIE2/angiopoietin in the angiogenesis and hematopoiesis]. PMID- 10846449 TI - [Prognostic factors in chemotherapies for acute myeloid leukemia]. PMID- 10846450 TI - [Anti-phospholipid antibody related immune thrombosis]. PMID- 10846451 TI - [Autoantigens in autoimmune hemolytic anemia]. PMID- 10846452 TI - [Changing concept of conventional therapy in chronic myelogenous leukemia]. PMID- 10846453 TI - [Monitoring of Ph chromosome by fluorescence in situ hybridization in chronic myeloid leukemia]. PMID- 10846454 TI - [Interferon-alpha therapy in chronic myelogenous leukemia]. PMID- 10846455 TI - [Extramedullary lymphoproliferative disorders in a patient with Ph-positive chronic myelogenous leukemia]. PMID- 10846456 TI - [Stem cell transplantation for chronic myelogenous leukemia]. PMID- 10846457 TI - [Donor leukocyte transfusion for relapsed chronic myeloid leukemia after allogeneic bone marrow transplantation]. PMID- 10846458 TI - [Vaccination of chronic myelogenous leukemia patients with peptide-pulsed dendritic cells]. PMID- 10846459 TI - [Retrospective study of acute myelogenous leukemia in 83 elderly patients: clinical and biological characteristics]. AB - In order to characterize clinical and biological characteristics of elderly patients with acute myelogenous leukemia (AML), we retrospectively analysed 83 elderly patients aged 60 years or more and, as a control, 114 younger patients aged 15 to 59 years who were admitted to our hospital between August 1984 and January 1998. There was a significantly higher incidence of preceding myelodysplastic syndromes in the elderly patients. They also had a significantly higher incidence of unfavorable cytogenetic abnormalities (loss or partial deletion of chromosome 5 or 7) and a significantly lower incidence of favorable cytogenetic abnormalities, such as t(15:17), t(8:21), or inv(16). With regard to FAB subtypes in de novo AML, the incidence of M3 subtype was significantly lower in the elderly group. Myeloperoxidase positivity of AML cells in the elderly group was lower than that in the younger group. Laboratory data at presentation disclosed a lower peripheral leukemic cell count, a higher fibrinogen level, a lower serum protein level, and a higher serum creatinine level in the elderly group. They also had poorer performance status and more frequent concomitant diseases at presentation, including liver diseases, heart diseases, or documented infections. It was concluded that elderly AML patients 60 years or older had a higher incidence of poor prognostic factors compared to younger patients. PMID- 10846461 TI - [Pulmonary complications after allogeneic hematopoietic stem cell transplantation]. AB - We investigated the occurrence of pulmonary complications in patients who underwent allogeneic hematopoietic stem cell transplantation at our institution. Pulmonary complications were observed in 12 out of 60 patients. Interstitial pneumonia developed in 12 cases: 7 idiopathic, 2 cytomegalovirus-associated, 1 P. carinii, 1 HSV, and 1 HHV-6-associated. HSV- and HHV-6-associated pneumonias were exhibited 100 days after transplantation. PCR analysis was diagnostically useful for detection of viral DNA in bronchial alveolar lavage fluid. Respiratory disease with airway obstruction was observed in 4 patients with chronic graft versus-host disease, and all 4 had a history of interstitial pneumonia. Three patients died of respiratory failure. Mycobacicrium avium complex was detected in 2. Exacerbation of respiratory failure may be associated with mycobacterial infection. PMID- 10846460 TI - [Retrospective study of acute myelogenous leukemia in elderly patients: treatment and outcome of 83 consecutive patients]. AB - We retrospectively analyzed treatments and outcomes for 83 acute myelogenous leukemia (AML) patients aged 60 years or more (median age 71) admitted to our hospital between August 1984 and January 1998. Complete remission was achieved in 36% of 78 patients who received anti-leukemic therapy, and median overall survival was 227 days. In addition to abnormal karyotypes involving chromosome 5 or 7, administration of less than 120 mg/m2/course of daunorubicin (DNR) during the initial treatment phase was an unfavorable prognostic factor for both CR and survival. Only 41% of all patients received 120 mg/m2/course of DNR or more, and had a significantly higher CR rate (56%) and longer survival, with a median of 389 days. It was suggested that intensive chemotherapy was effective for selected elderly AML patients who were relatively younger and had good performance status, although the number of such patients was limited in our study. PMID- 10846462 TI - [Evaluation of hematopoiesis by granulocyte elastase after hematopoietic stem cell transplantation]. AB - To evaluate hematopoietic restoration after hematopoietic stem cell transplantation (HSCT), we sequentially monitored the post-HSCT level of granulocyte elastase (GE), a sensitive parameter of qualitative and quantitative changes in granulopoiesis. We compared it with routinely used hematopoietic recovery indices, such as leukocyte and reticulocyte count. We also compared levels of GE in the clinical administration of different colony stimulating factors (granulocyte-colony stimulating factor (G-CSF) versus macrophage-colony stimulating factor (M-CSF) and in different types of hematopoietic stem cell transplantation (allogeneic bone marrow transplantation (BMT) versus autologus peripheral blood stem cell transplantation (PBSCT)). Days to first increase of GE after HSCT were 3.0 and 2.3 days earlier than increase of leukocytes in allo-BMT and auto-PBSCT, respectively. Recovery of highly fluorescent reticulocytes and monocytes were later than recovery of GE. These results indicated that granulopoiesis after transplantation started before the increase in peripheral leucocyte count, and that GE was the earliest indicator of hematopoietic recovery. On the basis of GE level, M-CSF had the same stimulating effect on granulopoiesis as G-CSF. The nadir of GE in PBSCT was significantly higher than that in BMT, indicating continuous granulopoiesis in PBSCT. From these results, we concluded that measurements of GE can be used for the clinical evaluation of myelosuppression by different conditioning regimens as well as of granulopoiesis induced by various cytokines. PMID- 10846463 TI - [Reactivation of hepatitis B virus after autologous peripheral blood stem cell transplantation in patients with positive hepatitis B surface antibodies]. AB - Hepatitis B virus (HBV) reactivation in patients with positive hepatitis B surface antibody (HBsAb) has been reported in some cases of allogenic bone marrow transplantation, acquired immunodeficiency syndrome (AIDS), and organ transplantation. However, to our knowledge, no reports have been made on the frequency and risk factors involved in HBV reactivation after autologous peripheral blood stem cell transplantation (APBSCT). Forty seven patients who underwent APBSCT were retrospectively analyzed. Three patients who were HBsAb positive before APBSCT contracted post-transplant HBV acute hepatitis. All 3 patients had multiple myeloma. HBV DNA could not be demonstrated in preserved samples of transfused blood. Therefore, we speculated that reactivation of latent HBV had occurred. The 3 patients with HBV hepatitis had relatively lower titers of pre-transplant HBsAb, and the total dose of steroids they received after APBSCT was significantly higher than for other patients who did not experience post-transplant HBV reactivation. There were no significant differences in pre transplant hepatitis B core antibody (HBcAb) titer or total post-transplant blood transfusion volume. Our study suggested that immunocompromised states, especially those induced by high-dose steroid therapy, may allow the reactivation of HBV after APBSCT, even in patients who had HBsAb before APBSCT. PMID- 10846464 TI - [Burkitt's lymphoma occurring as a primary lymphomatous effusion]. AB - A 39-year-old man was admitted with massive ascites. Specimens of ascitic fluid contained numerous cells with a FAB-L3 appearance, and small noncleaved cell lymphoma morphology. These cells expressed CD10, CD19, CD20, CD38, CD45, HLA-DR, and IgM antigens, and were positive for IgM and c-myc protein in cytoplasmic immunostaining tests. Clonal rearrangements of IgH and c-myc genes were detected by Southern blot analysis. No mass lesions were found by physical examination, and systemic computed photography did not reveal enlargement of lymph nodes, spleen, or liver. Bone marrow aspiration showed no infiltration of malignant cells. Ga scintigraphy indicated hot lesions only in the abdomen. These findings suggested that Burkitt's lymphoma had developed in the peritoneal cavity as a primary lymphomatous effusion. Chemotherapy with methotrexate, cyclophosphamide, vincristine, doxorubicin, etoposide, and dexamethasone was effective, and the patient has been free from the disease for 1 year since completion of consolidation treatment with autologous peripheral blood stem cell transplantation. PMID- 10846465 TI - [Undifferentiated blastic cell crisis of chronic myelogenous leukemia with myeloblastic tumor in the skin]. AB - A 54-year-old female, who had been treated for 4 years in the chronic phase of chronic myelogenous leukemia (CML) was admitted for management of a CML blastic crisis. Blast cells showed strong positive expression of CD7 and HLA-DR, and weakly expressed CD2, CD5 and CD10, as well. The cells were peroxidase negative in peripheral blood and bone marrow. An undifferentiated blastic crisis was diagnosed and she was treated with Interferon-alpha and VP(vincristine 2 mg/week; prednisolone 30 mg/day). A 5-7 mm in diameter tumor in the skin of the anterior right chest appeared one week after VP therapy. The tumor consisted of blasts which were CD13, CD33 and peroxidase positive, unlike the peripheral undifferentiated blasts. This is a rare case of mixed blast crisis with an increase in undifferentiated blasts in peripheral blood and bone marrow, and myeloblastic tumor formation in the skin. PMID- 10846466 TI - [High molecular-weight Bence Jones protein deposits in the kidney of a patient with plasma cell dyscrasia]. AB - A 70-year-old man was admitted to our hospital because of edema of the face and legs, proteinuria, and hypoproteinemia. We made a diagnosis of nephrotic syndrome. Immunoelectrophoresis of the patient's serum and urine disclosed a small amount of kappa type Bence-Jones protein (BJP) in the fast-gamma area. A renal biopsy showed nodular expansion of mesangium with deposition of space kappa light chain in the mesangial area. Light chain deposition disease was diagnosed. Western blotting analysis disclosed that the patient's BJP molecules had a molecular mass of 66 KDa, and were composed of two kappa chains of 33 KDa. Bone marrow aspiration revealed dysplastic plasma cells, and Western blotting analysis of an extract of these cells detected BJP-kappa with a molecular mass of 33 Kda. This BJP was larger than normal, indicating the possibility of abnormal structure. Structural abnormalities may be responsible for tissue precipitation of the kappa light chain. PMID- 10846467 TI - [Transformation of severe aplastic anemia to myelodysplastic syndrome with monosomy 7 in a patient who achieved transfusion independence after immunosuppressive therapy]. AB - A 72-year-old woman was given a diagnosis of severe aplastic anemia, and treated with anabolic steroid and cyclosporin A starting in October 1996. Because this treatment was ineffective, anti-thymocyte globulin (ATG) therapy was started in September 1997. In May 1998, chromosome analysis revealed transformation to myelodysplastic syndrome (MDS), refractory anemia with excess of blasts with monosomy 7 in 60% of metaphase cells. The patient showed gradual hematologic improvement and became transfusion independent. Despite progression to acute myeloid leukemia (FAB-M6) with monosomy 7 in 100% of metaphase cells in December 1998, the hemoglobin level recovered to 13.2 g/dl. In May 1999 the blasts increased rapidly and transformation to acute myelomonocytic leukemia (FAB-M4) was diagnosed. The patient was treated with low-dose Ara-C and aclarubicin with no improvement and died in August 1999. This case demonstrated the transformation of severe aplastic anemia to acute myeloid leukemia via MDS with monosomy 7 associated with transfusion independence after immunosuppressive therapy. These findings suggested a close relationship between aplastic anemia and hypoplastic MDS and the possibility of hematologic improvement based on the growth advantage of abnormal clones. PMID- 10846468 TI - [Successful treatment of relapsed and refractory acute promyelocytic leukemia with arsenic trioxide (As2O3)]. AB - It has been shown that arsenic trioxide (As2O3) may induce hematologic remissions in patients with acute promyelocytic leukemia (APL) refractory to all-trans retinoic acid (ATRA). We reported on a patient with ATRA and drug-resistant APL that was successfully treated with As2O3. The patient had been given a diagnosis of typical APL and was treated with ATRA and chemotherapy for 12 months. He achieved complete remission (CR), but leukemia relapsed with 43% APL cells in the bone marrow in the 16th month of treatment. ATRA and cytarabine plus daunorubicin were administered; however, the APL cells in the bone marrow increased to 97.2%. As2O3 was initiated intravenously, and bone marrow showed a decrease of APL cells (6.7%) and a partial differentiation after 9 days. The patient received idarubicin (IDA) and steroid pulse because of the development of ATRA-like syndrome, and achieved CR 37 days after the initiation of As2O3. He received an additional 2 courses of As2O3 with IDA, and is in CR. These results demonstrated the therapeutic efficacy of As2O3 in treating ATRA and drug-resistant APL. PMID- 10846469 TI - [Molecular mechanism of neuronal survival and regeneration after injury]. PMID- 10846470 TI - [The origin and evolution of chloroplasts]. PMID- 10846471 TI - [The mechanism of vancomycin-resistant in Staphylococcus aureus]. PMID- 10846472 TI - [Molecular biology of enzymes degrading pollutants: diversity and convergent evolution observed on extradiol dioxygenases]. PMID- 10846473 TI - [The cellular target proteins of adenovirus gene products]. PMID- 10846474 TI - [Information from the complete genome sequence of Chlamydia trachomatis: an obligate intracellular pathogen of humans]. PMID- 10846475 TI - [The complete genome sequence of Rickettsia prowazekii, the etiological agent of epidemic typhus: significance as pathogen and symbiont]. PMID- 10846476 TI - [Tissue engineering: cooperation of molecular biology and polymer science]. PMID- 10846477 TI - [High magnetic-field high-pressure NMR facility at Kobe University]. PMID- 10846478 TI - [Mass spectrometry of a protein (3): Ion trap mass spectrometer and its application]. PMID- 10846479 TI - [Scientific pedagogical activity at the St. Petersburg ENT and Speech Research Institute]. PMID- 10846480 TI - [Rehabilitation and assessment of aural-oral speech development in children with cochlear implants]. AB - Updated models of cochlear implants provide good speech audibility and thus complete rehabilitation of children who have lost hearing after learning speech. All the children who lost hearing before learning speech can hear sounds of normal loudness and orient in sound media by means of cochlear implant. However, they need long-term audio-vocal rehabilitation the results of which depend on the age of the child's operation and hi(her) individual traits. Cochlear implants in children aged under 3 years are most perspective. Russian language methodology including 7 tests and 2 questionnaires is described. It is intended for assessment of audio-vocal development in children with cochlear implants and results of audio-vocal rehabilitation as well as of effectiveness of using cochlear implant in children over 2 years of age. Establishment of centers for cochlear implantation and introduction of cochlear implantation state program are recommended. PMID- 10846481 TI - [The use of computer electronystagmography in the assessment of optokinetic nystagmic reactions]. AB - A novel technique of computer-assisted electronystagmography (CENG) has been used in 90 patients with vertigo. Two variants of optokinetic nystagmus (cortical and subcortical) were analysed. Registration of optokinetic nystagmus (OKN) was carried out using original devices for CENG. Right- and left-directed nystagmic reactions and their quantitation were performed according to specially devised registering and analysing programs. In one-sided vestibular disorders cases of cross-over OKN asymmetry prevailed in affection of the cerebellopontine angle (neurinoma of the VIII nerve) compared to labyrinthine disturbances (one-sided Meniere's disease). In 90% of patients with vertebrobasillar deficiency OKN asymmetry was found even in a straight position of the head. OKN asymmetry was less pronounced or disappeared in the OKN test performed with the head turned right or left by 90 degrees. PMID- 10846482 TI - [Adrenergic innervation of the middle and internal ear in man]. AB - Adrenergic nervous structures (ANS) in morphofunctional formations of human middle and internal ear were studied on the autopsy material obtained from 5 victims of accidents free of any ear disease. The samples were treated histochemically with glyoxylic acid. Non-varicose dilated sympathetic nerve fibers (single and in bundles, non-myelinized, intensively luminescent, twisted and direct) were detected in vertical nerve sections, in spiral and vestibular ganglia, in chorda tympani. A thick net of adrenergic nerve fibers (ANF) was noted around the labyrinthine artery. ANF were absent long along the walls of the utriculus and semicircular canals, in the modiolus vessels, spiral ligament, vascular stripe. ANF were most numerous on the osseous spiral lamina. Near the Corti's organ ANF are thicker and often varicose. On the vertical sections of m. stapedius ANF with varicose parts locate in the connective tissue perimysial layers and in the endomysium directly between the muscular cells. At the site of muscle transition into the tendon and in tendon as such no ANF were found. Two sets of ANF were noted in the middle ear mucosa: perivascular and unrelated to blood vessels. PMID- 10846483 TI - [Experience in the early diagnosis and habilitation of children with hearing loss in St. Petersburg]. AB - By estimation of the specialists, the present system of early detection of hearing problems in children provides registration of hypoacusis only in 40% of hypoacusis infants. To improve the situation, it is necessary to update the screening system, to integrate efforts of various specialists, to perfect information support, training of specialists, provide modern hearing aids, organize rehabilitation groups including both normal children and those with hearing and speech problems. PMID- 10846484 TI - [Specific aspects of the reflection of signal frequency structure in the peripheral compartment of acoustic analyzer in health state and disease]. AB - How frequency structure of the signals is reflected at the exit of the model of acoustic system peripheral compartment was investigated. The system incorporated mechanisms of local feedback by microphone and overall potentials. Defects in the work of the feedback mechanisms lead to substantial deformation of sound reflection at the model exit. Format structure of the sounds is not reflected. This deteriorates perception. To restore the reflection of the format structure, previous reconstruction is proposed consisting in enhanced degree of contrast of the frequency signal structure. PMID- 10846485 TI - [Morpho-functional status of the pharyngeal and palatine tonsils in children with regional lymphadenitis]. AB - The authors give clinicomorphological analysis of 100 cases of chronic tonsillitis in young children who had marked regional lymphadenitis in the absence of clinical manifestations of tonsillitis. General and local immunity were examined in these children. Lifetime assessment was made of functional tonsil condition by cytological and microbiological examination of the content of deep lacune parts. Removed palatine tonsils were examined histologically, immunohistochemically and ultrastructurally. Uncommon location of the upper tonsillar pole impairs drainage function of the lacunes, attenuation of lymphoepithelial symbiosis, functional activity of the affected part of the tonsils which give rise to pathological processes in the regional lymph nodes. Impairment of lymph nodes becomes leading in clinical course of the disease. PMID- 10846486 TI - [Adrenergic innervation of nasal polyps and pharyngeal tonsil in children]. AB - Vegetative nervous structures in adenoids and nasal polyps were studied histochemically. Functional parameters of the vegetative nervous system were evaluated in children with hyperplasia of the pharyngeal tonsil. Polyps are shown to have adrenergic nervous structures. 80% of tonsillar hyperplasia children exhibited vegetative dystonia. Adrenergic innervation of adenoids removed surgically is described. PMID- 10846487 TI - [Ultrastructure of nasal and paranasal mucosa in patients with chronic polypous rhinosinusitis]. AB - Electron microscopic study of operative samples of nasal and paranasal mucosa from patients with chronic polypous rhinosinusitis has discovered changes in the mucosa typical for inflammation and ultrastructural changes. Epithelial layer underwent destruction. Adequate function of the remaining ciliary cells is doubtful even in minimal changes of their ciliary apparatus. The absence of the specialized contacts between the epithelyocytes indicates dysfunction of the selective ionic permeability. There was overgrowth of the reticular fibers of the basal membrane. Plasma cells exhibited marked dilation of the cysterns of the rough endoplasmic reticulum probably because of active accumulation of immunoglobulins or their precursors. PMID- 10846488 TI - [Sphenoid sinuses in preschool children in health and disease]. AB - The study of computer and MR tomograms of the paranasal sinuses in preschool children provided information of pneumatization of the sphenoid sinuses, their shape and size. 17 children had clinical and x-ray picture of sphenoiditis. Therapeutic measures in acute sphenoiditis in children are proposed. PMID- 10846489 TI - [Present day diagnosis of voice disorders in children]. AB - Videoendostroboscopy (VESS) was employed for diagnosis of vocal dysfunction in 369 children aged 6 to 16 years. Effects of flexible and hard endoscopes were compared. Vibratory cycle was analyzed in 55 healthy children of different age. Diagnosis and differential diagnosis of soft and hard nodules, physiological and long-term mutation, chronic laryngitis are described as well as a videoendostroboscopic picture in functional dysphonias, vocal grooves and polyps. VESS is a modern, informative and safe diagnostic procedure. PMID- 10846490 TI - [Effect of recombinant interleukin-1 beta on microbial flora of the middle ear in patients with chronic purulent otitis media]. AB - Betaleukin was given to 60 patients with various forms of otitis media purulenta chronica (OMPC). Symptoms of the purulent exacerbation were relieved in 43.3% of the patients, the clinical course improved in 18.3%. No response was achieved in 40% of the treated patients. Betaleukin proved highly effective in management of exacerbations of uncomplicated OMPC though it has no direct antimicrobial activity. PMID- 10846491 TI - [Functional status of the autonomic nervous system in patients with polyps of the vocal cords]. AB - Assessment of functional state of the autonomic nervous system (tonicity, reactivity, support of the cardiovascular system) was made in 34 patients aged 18 64 years admitted to the ENT Research Institute for removal of laryngeal tumors. All the patients were diagnosed to have polyps of the vocal folds. Comprehensive studies suggest the conclusion on morphofunctional changes in the vocal folds which combine with adaptation-trophic disorders in the autonomic nervous system. PMID- 10846492 TI - [Band filtration of the speech signal: speech perception in health status and in sensorineural hearing loss]. AB - We made band speech filtration by dividing the spectrum into the frequency bands 50, 100, 200, 500 and 1000 Hz with following removal of the even or odd bands. We found that audibility of speech in all the examinees with normal hearing was 100% irrespective of the band width and composition of the speech signal (a total of the even or odd bands). Audibility of speech composed of odd bands 100 Hz width improved by 4 to 16% in patients with neurosensory hypoacusis. PMID- 10846493 TI - [Observation of congenital middle ear malformation in a 14-year-old child]. PMID- 10846494 TI - [Chordoma of the temporal bone]. PMID- 10846496 TI - Time for a national agenda to improve the health of urban populations. PMID- 10846495 TI - [The role of mucous membranes in ENT defense from potentially pathogenic antigenic factors]. PMID- 10846497 TI - Getting political: racism and urban health. PMID- 10846498 TI - The neglected epidemic and the surgeon general's report: a call to action for better oral health. PMID- 10846499 TI - Water quality laws and waterborne diseases: Cryptosporidium and other emerging pathogens. AB - Waterborne diseases, such as cryptosporidiosis, cause many cases of serious illness in the United States annually. Water quality is regulated by a complex system of federal and state legal provisions and agencies, which has been poorly studied. The authors surveyed state and territorial agencies responsible for water quality about their laws, regulations, policies, and practices related to water quality and surveillance of cryptosporidiosis related to drinking water. In this commentary they review the development and current status of federal drinking water regulations, identify conflicts or gaps in legal authority between federal agencies and state and territorial agencies, and describe court-imposed limitations on federal authority with regard to regulation of water quality. Recommendations are made for government actions that would increase the efficiency of efforts to ensure water quality; protect watersheds; strengthen waterborne disease surveillance; and protect the health of vulnerable populations. PMID- 10846500 TI - Small taxes on soft drinks and snack foods to promote health. AB - Health officials often wish to sponsor nutrition and other health promotion programs but are hampered by lack of funding. One source of funding is suggested by the fact that 18 states and 1 major city levy special taxes on soft drinks, candy, chewing gum, or snack foods. The tax rates may be too small to affect sales, but in some jurisdictions, the revenues generated are substantial. Nationally, about $1 billion is raised annually from these taxes. The authors propose that state and local governments levy taxes on foods of low nutritional value and use the revenues to fund health promotion programs. PMID- 10846501 TI - Community, service, and policy strategies to improve health care access in the changing urban environment. AB - Urban communities continue to face formidable historic challenges to improving public health. However, reinvestment initiatives, changing demographics, and growth in urban areas are creating changes that offer new opportunities for improving health while requiring that health systems be adapted to residents' health needs. This commentary suggests that health care improvement in metropolitan areas will require setting local, state, and national agendas around 3 priorities. First, health care must reorient around powerful population dynamics, in particular, cultural diversity, growing numbers of elderly, those in welfare-workplace transition, and those unable to negotiate an increasingly complex health system. Second, communities and governments must assess the consequences of health professional shortages, safety net provider closures and conversions, and new marketplace pressures in terms of their effects on access to care for vulnerable urban populations; they must also weigh the potential value of emerging models for improving those populations' care. Finally, governments at all levels should use their influence through accreditation, standards, tobacco settlements, and other financing streams to educate and guide urban providers in directions that respond to urban communities' health care needs. PMID- 10846502 TI - Urban issues in health promotion strategies. AB - The powerful influence of behavioral choices on health status is well established. The implications and challenges for urban populations are formidable. Understanding urban environments will better prepare health promotion professionals to deal effectively with the forces affecting health-related behaviors. In thinking about urban health promotion in the United States, researchers often distinguish between 2 frameworks; one contending with urbanization, which affects most of us, and another contending with inner-city environments, where many of the deepest needs are. Urbanization confers both benefits and liabilities, but the single greatest challenge for health promotion may lie in reestablishing positive social connections. In contrast, 2 key features of the inner-city environment may be the negative ecological forces within neighborhoods and the lack of control over one's fate. Too often, prescriptions for the inner city stereotype its problems and ignore its strengths. For the inner city, important foundation stones for the future include ways to build on these strengths through positive connections and increased community control through coalition building. PMID- 10846504 TI - Breath of Life: stories of asthma from an exhibition at the National Library of Medicine. PMID- 10846503 TI - To mitigate, resist, or undo: addressing structural influences on the health of urban populations. AB - Young to middle-aged residents of impoverished urban areas suffer extra-ordinary rates of excess mortality, to which deaths from chronic disease contribute heavily. Understanding of urban health disadvantages and attempts to reverse them will be incomplete if the structural factors that produced modern minority ghettos in central cities are not taken into account. Dynamic conceptions of the role of race/ethnicity in producing health inequalities must encompass (1) social relationship between majority and minority populations that privilege the majority population and (2) the autonomous institutions within minority populations that members develop and sustain to mitigate, resist, or undo the adverse effects of discrimination. Broad social and economic policies that intensify poverty or undermine autonomous protections can reap dire consequences for health. Following from this structural analysis and previous research, guiding principles for action and suggestions for continued research are proposed. Without taking poverty and race/ethnicity into account, public health professionals who hope to redress the health problems of urban life risk exaggerating the returns that can be expected of public health campaigns or overlooking important approaches for mounting successful interventions. PMID- 10846505 TI - Dr Louis T. Wright and the NAACP: pioneers in hospital racial integration. AB - Louis Tompkins Wright, the son of a man born into slavery, was an outstanding African American surgeon who devoted his life to the racial integration of health care in the United States. Despite the fact that both his father and stepfather were physicians, despite his innate intellectual gifts and disciplined character, Wright experienced discrimination throughout his life and career. This experience led him to fight for the rights of African Americans, both health care professionals and patients. In addition to making numerous contributions in the fields of surgery and infectious disease, Wright held leadership positions in the National Association for the Advancement of Colored People for more than 20 years, leaving a legacy of equity for African Americans in medical education and in health care. PMID- 10846506 TI - For the welfare of children: the origins of the relationship between US public health workers and pediatricians. AB - The majority of children living in the United States today enjoy excellent health and access to health care. This was not always so; before the late 19th century, the field of pediatric medicine scarcely existed, and the combination of harsh and unsanitary living conditions in the urban areas where most immigrants settled, infectious diseases, and improper handling of milk was particularly deadly for infants and children. This article discusses the relationship between pediatric medicine and the broader children's health and public health movements in the United States in the early decades of the 20th century. That relationship resulted in 3 developments that had a profound impact on children's health: the establishment of dispensaries and milk stations that served impoverished children, campaigns to educate parents about illness prevention and child rearing, and the medical inspection of public schools and schoolchildren. Today, American children face both new threats to health and the reemergence of infectious diseases that were once thought conquered. Pediatricians and public health professionals must work together in the same spirit of social activism and community responsibility to meet these challenges. PMID- 10846507 TI - State welfare reform policies and declines in health insurance. AB - OBJECTIVES: This study sought to determine whether there is a relationship between state policies on Temporary Assistance to Needy Families (TANF), declines in both TANF and Medicaid caseloads, and the rise in the number of uninsured. METHODS: Extant data sources of state TANF policies, TANF and Medicaid participation, and uninsurance rates were analyzed, with the state as the unit of analysis. The independent variables included state TANF policies that directly address receipt of benefits or relate to health; dependent variables included changes in state TANF enrollment, Medicaid enrollment, and health insurance status since the enactment of the law. RESULTS: In the bivariate analysis, declines in Medicaid were associated with sanction for work noncompliance, lack of a child care guarantee, and strategies to deter TANF enrollment; this last factor was also associated with increased uninsurance. In the multivariate analysis, lack of a child care guarantee and deterrent strategies predicted TANF declines; deterrent strategies predicted Medicaid decline and uninsurance increases. CONCLUSIONS: This analysis suggests that policies deterring TANF enrollment may contribute to declines in Medicaid and increased uninsurance. To maintain health insurance for the poor, policymakers should consider revising policies that deter TANF enrollment. PMID- 10846508 TI - Gender differences in health care access indicators in an urban, low-income community. AB - OBJECTIVES: This study examined factors associated with gender differences in health insurance coverage and having a usual source of medical care. METHODS: In person interviews were conducted with a community sample of 695 residents of Central Harlem, New York City. Predictors of the 2 outcome variables and the interaction of key variables with gender were analyzed via logistic regression. RESULTS: No strong patterns emerged to explain gender differentials in having insurance coverage and having a usual provider. However, women employed full time had increased odds of insurance coverage, whereas employment had no similar effect among men. Public assistance evidenced a strong relationship with insurance coverage among both men and women. Socioeconomic factors and health insurance were important independent predictors of having a usual source of health care for men but had little effect among women. CONCLUSIONS: Expanding the availability of both public insurance and affordable private coverage for men living in low-income communities is an important means of reducing gender disparities in access to health care. Public assistance is an important means of enabling access to health care for men as well as women. PMID- 10846509 TI - Health insurance coverage of immigrants living in the United States: differences by citizenship status and country of origin. AB - OBJECTIVES: This study examined health insurance coverage among immigrants who are not US citizens and among individuals from the 16 countries with the largest number of immigrants living in the United States. METHODS: We analyzed data from the 1998 Current Population Survey, using logistic regression to standardize rates of employer-sponsored coverage by country of origin. RESULTS: In 1997, 16.7 million immigrants were not US citizens. Among non-citizens, 43% of children and 12% of elders lacked health insurance, compared with 14% of non-immigrant children and 1% of non-immigrant elders. Approximately 50% of non-citizen full time workers had employer-sponsored coverage, compared with 81% of non-immigrant full-time workers. Immigrants from Guatemala, Mexico, El Salvador, Haiti, Korea, and Vietnam were the most likely to be uninsured. Among immigrants who worked full-time, sociodemographic and employment characteristics accounted for most of the variation in employer health insurance. For Central American immigrants, legal status may play a role in high un-insurance rates. CONCLUSIONS: Immigrants who are not US citizens are much less likely to receive employer-sponsored health insurance or government coverage; 44% are uninsured. Ongoing debates on health insurance reform and efforts to improve coverage will need to focus attention on this group. PMID- 10846510 TI - What people really know about their health insurance: a comparison of information obtained from individuals and their insurers. AB - OBJECTIVES: This study determined the validity of self-reported data on selected health insurance characteristics. METHODS: We obtained telephone survey data on the presence of health insurance, source of insurance, length of time insured, and type of insurance (managed care or fee-for-service) from a random sample of 351 adults in 3 Wisconsin counties and compared findings with data from respondents' health insurers. RESULTS: More than 97% of the respondents correctly reported that they were currently insured. For source of insurance among persons aged 18 to 64 years, sensitivity was high for those covered through private health insurance (93.8%) but low for those covered through public insurance (6.7%). Only 33.1% of the respondents accurately categorized length of enrollment in their current plan. Overall estimates for managed care enrollment were similar for the 2 sources, but individual validity was low: 84.2% of those in fee-for service believed that they were in managed care. CONCLUSIONS: Information obtained from the general population about whether they have health insurance is valid, but self-reported data on source of insurance, length of time insured, and type of insurance are suspect and should be used cautiously. PMID- 10846511 TI - Asthma and Latino cultures: different prevalence reported among groups sharing the same environment. AB - OBJECTIVES: This 1999 study measured asthma prevalence among Latinos of different cultural traditions who live on the same streets and in the same buildings. METHODS: Health promoters from El Puente in North Brooklyn, New York City, surveyed 3015 people in 946 households, asking standard asthma prevalence questions. RESULTS: Some 46% of households identified themselves as Dominican, 42% as Puerto Rican, 6% as other Latino, and 6% as other. Reported asthma period prevalence was 5.3% (93 of 1749) among Dominicans and other Latinos, compared with 13.2% (147 of 1115) among Puerto Ricans (odds ratio = 0.37; 95% confidence interval = 0.28, 0.49), a difference not explained by location (cluster or building), household size, use of home remedies, educational attainment, or country where education was completed. Differences were least detectable among 13 to 24-year-olds of both sexes and sharpest among women aged 45 years and older and girls from birth to 12 years. CONCLUSIONS: Further research on gene environment interactions is needed among Puerto Ricans and Dominicans, but asthma's associations with low income and unhealthy environment, which more recent immigrants seem better able to withstand, should not be overlooked. PMID- 10846512 TI - The effect of a needle exchange program on numbers of discarded needles: a 2-year follow-up. AB - OBJECTIVES: This study estimates the quantity and geographic distribution of discarded needles on the streets of Baltimore, Md, during the 2 years after a needle exchange program opened. METHODS: Thirty-two city blocks were randomly sampled. Counts were taken of the number of syringes, drug vials, and bottles before the needle exchange program opened and then at 6 periodic intervals for 2 years after the program opened. Nonparametric and generalized estimating equation models were used to examine change over time. RESULTS: Two years after the needle exchange program opened, there was a significant decline in the overall quantity of discarded needles relative to that of drug vials and bottles (background trash). The block mean of number of needles per 100 trash items was 2.42 before the program opened and 1.30 2 years later (mean within-block change = -0.028, P < .05). There was no difference in the number of discarded needles by distance from the program site. CONCLUSIONS: These data suggest that this needle exchange program did not increase the number of distribution of discarded needles. PMID- 10846513 TI - Helping women quit smoking: results of a community intervention program. AB - OBJECTIVES: This intervention was implemented to reduce the prevalence of cigarette smoking among women. METHODS: We used community organization approaches to create coalitions and task forces to develop and implement a multicomponent intervention in 2 counties in Vermont and New Hampshire, with a special focus on providing support to help women quit smoking. Evaluation was by pre-intervention and post-intervention random-digit-dialed telephone surveys in the intervention counties and the 2 matched comparison counties. RESULTS: In the intervention counties, compared with the comparison counties, the odds of a woman being a smoker after 4 years of program activities were 0.88 (95% confidence interval = 0.78, 1.00) (P = .02, 1-tailed); women smokers' perceptions of community norms about women smoking were significantly more negative (P = .002, 1-tailed); and the quit rate in the past 5 years was significantly greater (25.4% vs 21.4%; P = .02, 1-tailed). Quit rates were significantly higher in the intervention counties among younger women (aged 18 to 44 years); among women with household annual incomes of $25,000 or less; and among heavier smokers (those who smoked 25 or more cigarettes daily). CONCLUSIONS: In these rural counties, community participation in planning and implementing interventions was accompanied by favorable changes in women's smoking behavior. PMID- 10846514 TI - Dietary supplements and physical exercise affecting bone and body composition in frail elderly persons. AB - OBJECTIVES: This study determined the effect of enriched foods and all-around physical exercise on bone and body composition in frail elderly persons. METHODS: A 17-week randomized, controlled intervention trial, following a 2 x 2 factorial design--(1) enriched foods, (2) exercise, (3) both, or (4) neither--was performed in 143 frail elderly persons (aged 78.6 +/- 5.6 years). Foods were enriched with multiple micronutrients; exercises focused on skill training, including strength, endurance, coordination, and flexibility. Main outcome parameters were bone and body composition. RESULTS: Exercise preserved lean mass (mean difference between exercisers and non-exercisers: 0.5 kg +/- 1.2 kg; P < .02). Groups receiving enriched food had slightly increased bone mineral density (+0.4%), bone mass (+0.6%), and bone calcium (+0.6%) compared with groups receiving non-enriched foods, in whom small decreases of 0.1%, 0.2%, and 0.4%, respectively, were found. These groups differed in bone mineral density (0.006 +/- 0.020 g/cm2; P = .08), total bone mass (19 +/- g; P = .04), and bone calcium (8 +/- 21 g; P = .03). CONCLUSIONS: Foods containing a physiologic dose of micronutrients slightly increased bone density, mass, and calcium, whereas moderately intense exercise preserved lean body mass in frail elderly persons. PMID- 10846515 TI - Mobility impairments and use of screening and preventive services. AB - OBJECTIVES: Primary care for people with disabilities often concentrates on underlying debilitating disorders to the exclusion of preventive health concerns. This study examined use of screening and preventive services among adults with mobility problems (difficulty walking, climbing stairs, or standing for extended periods). METHODS: The responses of non-institutionalized adults to the 1994 National Health Interview Survey, including the disability and Healthy People 2000 supplements, were analyzed. Multivariable logistic regressions predicted service use on the basis of mobility level, demographic characteristics, and indicators of health care access. RESULTS: Ten percent of the sample reported some mobility impairment; 3% experienced major problems. People with mobility problems were as likely as others to receive pneumonia and influenza immunizations but were less likely to receive other services. Adjusted odds ratios for women with major mobility difficulties were 0.6 (95% confidence interval [CI] = 0.4, 0.9) for the Papanicolaou test and 0.7 (95% CI = 0.5, 0.9) for mammography. CONCLUSIONS: More attention should be paid to screening and preventive services for people with mobility difficulties. Shortened appointment times, physically inaccessible care sites, and inadequate equipment could further compromise preventive care for this population. PMID- 10846516 TI - The association between greater continuity of care and timely measles-mumps rubella vaccination. AB - OBJECTIVES: This study assessed whether greater continuity of care is associated with timely administration of measles-mumps-rubella (MMR) vaccination. METHODS: We studied 11,233 patients continuously enrolled in Group Health Cooperative (GHC) from birth to 15 months. We used a preestablished index to quantify continuity of care based on the number of primary care providers in relation to the number of clinic visits. MMR vaccination status at 15 months was assessed with automated immunization data systems at GHC. RESULTS: In a logistic regression model, both medium continuity (odds ratio [OR] = 1.20, 95% confidence interval [CI] = 1.08, 1.33) and high continuity (OR = 1.36, 95% CI = 1.22, 1.52) were associated with increased likelihood of being immunized by 15 months compared with patients in the lowest tercile of continuity of care. CONCLUSION: Greater continuity of care is associated with more timely immunization. PMID- 10846517 TI - Menopausal hormones and breast cancer in a biracial population. AB - OBJECTIVES: This study examined the association between menopausal hormones and breast cancer in a biracial population. METHODS: Logistic regression was used to calculate odds ratios for breast cancer associated with hormone use among 397 cases and 425 controls, all menopausal women. RESULTS: Odds ratios for ever use of hormones were 0.8 (95% confidence interval [CI] = 0.5, 1.2) for White women and 0.7 (95% CI = 0.4, 1.2) for Black women. Risk was not increased with longer duration of use or more recent use. CONCLUSIONS: Breast cancer risk was not increased among White or Black women who used menopausal hormones, despite patterns of use varying considerably between races. PMID- 10846518 TI - Acute hazards to young children from residential pesticide exposures. AB - OBJECTIVES: This study assessed acute hazards to young children from pesticides toxic enough to require child-resistant packaging. METHODS: The names of pesticides meeting acute toxicity criteria were ascertained from the Environmental Protection Agency. Poison Control Center reports identified children younger than 6 years who were exposed to these pesticides. Toxicity category, medical outcome, sex, and age were examined. RESULTS: A higher proportion of children with exposure to the more toxic products had serious medical outcomes. Children 2 years and younger were the predominant age group exposed. CONCLUSIONS: Protective measures--substituting less lethal pesticides, reducing the concentration of the active ingredients, and improving packaging and storage--are recommended. PMID- 10846519 TI - Prevalence of home pregnancy testing among adolescents. AB - OBJECTIVES: This study estimated the prevalence of home pregnancy testing among adolescents. METHODS: A survey was administered in 11 urban clinics to 600 females aged 13 to 19 years. RESULTS: The prevalence of home pregnancy test use was 34% among 474 sexually experienced youths; 77% of the users had received at least 1 negative pregnancy test result, and 48% took no further action for confirmation. Compared with those who had never used such tests, users were older, younger at sexual debut, less likely to consistently use effective birth control, and more likely to have ever been pregnant. CONCLUSIONS: Health care clinics are important sources for pregnancy prevention, but clinics may have limited opportunity to intervene with some youths who use home pregnancy tests. PMID- 10846520 TI - Effects of after-school physical activity on fitness, fatness, and cognitive self perceptions: a pilot study among urban, minority adolescent girls. PMID- 10846521 TI - A reply to Sullivan's reanalysis of managed care plan performance since 1980. PMID- 10846522 TI - Misperceptions of "objective measurements"? PMID- 10846523 TI - [Embracing change]. PMID- 10846524 TI - [A current approach to the fight against cancer: "Specialty cancerology". The example of gynecological and breast oncology in regional hospital centers]. AB - The early diagnosis is the more efficient means to improve the overall survival of patients with cancer. The modern methods of diagnosis developed by specialists of cancer arising in a particular site, and by radiologists and pathologists, enable to more often diagnose precancerous lesions and cancer at an early stage. This small lesions can be cured by limited and conservative treatment. However, the diagnosis, the accurate evaluation and optimal treatment of these lesions require a perfect and specific knowledge of the pathology of a particular organ. According to the experience of gynecology and breast oncology, the author recommends guidelines for the diagnosis and the primary treatments of cancer proceeded in public hospitals. Patients care is managed in unit, including medical staff sub-specialized in oncology of a specific organ. This structure warrants the best quality of the diagnosis means and of the medical care at a lowest cost. PMID- 10846525 TI - [The organization of cancerology]. PMID- 10846526 TI - Analytical methods for evaluation on whole cells of human papillomavirus infection. AB - Analytical methods for evaluation on whole cells of human papillomavirus infection. Human papillomavirus (HPV) infection is currently identified by the presence of viral DNA using molecular biology. As in situ hybridization is valuable for HPV-DNA detection mainly with non-isotopic probes, we evaluated the sensitivity of various techniques, using as models three cell lines containing different copy numbers of HPV DNA/cell (CaSki with 600 copies of HPV 16, SiHa with 1-2 copies of HPV 16, HeLa with 10-50 copies of HPV 18). Epifluorescence microscopy and flow cytometry allowed detection of 600 copies in CaSki cells; in addition, cell fixation was found to influence the fluorescent intensity. Several procedures were assayed to increase the sensitivity of in situ hybridization. The use of biotinylated HPV-16 oligonucleotides as probes was not effective, because only CaSki cells were positive. After amplification of HPV-16 or -18 DNA sequences with polymerase chain reaction (PCR) on whole cells in suspension and hybridization with plasmid probes, fluorescent hybridization spots were found in CaSki and HeLa cells by both epifluorescence microscopy and flow cytometry. The various procedures applied for revelation of DNA-DNA hybrids (use of phycoerythrin or cyanine instead of fluorescein, Pinkel's 3-step amplified system of fluorescein) did not enhance the sensitivity of in situ hybridization. HPV DNA was very effectively detected by cell examination under a laser-scanning confocal microscope, since 1-2 copies of HPV 16 were observed in SiHa cells without previous PCR amplification. Thus, the efficacy of in situ hybridization for HPV detection may be conditioned by different factors. Laser-scanning microscopy represents an alternative to the use of PCR amplification. These techniques are potentially useful to study single genes. PMID- 10846527 TI - Flow cytometric evaluation of multidrug-resistance circumvention. AB - Flow cytometry, through the measurement of P-gp expression and function, is an important tool in the characterization of multidrug resistant cell lines used in pharmacology. Furthermore, as a complement to classical in vitro tests, this technique allows rapid evaluation of the capacity of new modulator agents to reverse the resistance to cytotoxic drugs and contributes a deeper insight into the mechanism of action of these new molecules. PMID- 10846528 TI - Immunophenotyping of acute leukemia by flow cytometry: closing the black holes of morphology. AB - The heterogeneity of disease conditions in acute leukemia (AL) requires the most discriminative procedures for a differential diagnostic classification. Selected cases of AL, representative of lineage fidelity, multilineage phenotype and phenotypic switch, are reviewed to document that the diversity of the leukemic cells (lineage involved, maturation stage, aberrant markers), although not always characterizable by morphological differences, is reflected by a variety of immunophenotypes which may be efficiently characterized by flow cytometry to obtain a precise diagnosis. PMID- 10846529 TI - Clinical flow cytometric DNA analysis of human solid tumors. AB - Flow cytometry is widely used to analyze nuclear DNA content. Its clinical application in oncology is becoming increasingly important for detecting abnormalities in DNA content (DNA aneuploidy) of cells from solid tumors and for monitoring cell-growth kinetics in tumor cells (S-phase estimation), information which may be relevant for prognosis and evaluation of therapy. The risks of false DNA ploidy determination and inaccurate S-phase estimation are discussed. A methodological approach, criteria for histogram interpretation and criteria for histogram rejection are proposed in order to avoid these artifactual results. PMID- 10846530 TI - [CD44 expression: a new prognostic factor in neuroblastoma]. AB - CD44 gene products are potential markers of aggressiveness in different tumor models, a result which prompted us to study clinical neuroblastoma (NB) specimens. CD44 expression was determined by immunostaining of 52 NB with a monoclonal antibody (J173) directed against an epitope common to all CD44 isoforms. All tumors were from patients (pts) with newly diagnosed NB treated with standardized protocols. They were classified according to international criteria [11]. CD44 immunoreactivity was detected in 37 tumors (71%). CD44 was expressed in 100% of favorable NB stages (1, 2 or 4S), but only 50% of advanced NB (stages 3 and 4) (p = 0.0001), suggesting that the absence rather that the overexpression of CD44 is a signal of tumor aggressiveness. The cumulative event free survival was significantly longer in pts with CD44-positive tumors as compared to pts with CD44-negative tumors (p < 10(-5)). More importantly, progression-free survival was also significantly higher in CD44-positive pts within the high-risk group (p < 0.01). In univariate analyses, we tested the prognostic value of tumor expression of CD44 in comparison with tumor stage, age, tumor histology and presence or absence of N-myc proto-oncogene amplification. All five measures had significant prognostic value. The expression of CD44 and the absence of N-myc amplification were the most powerful predictors of a favorable clinical outcome. In a multivariate analysis of these measures, CD44 expression and tumor stage were the only independent prognostic factors for the prediction of patient survival. NB is the first clinical model described in which tumor aggressiveness correlates with a repression rather than a stimulation of CD44 expression. We recommend the use of CD44 as an additional biological marker in the initial staging of neuroblastoma. PMID- 10846531 TI - [Initial locoregional chemotherapy in recurrent and locoregional advanced or inoperable stages of pelvic uterine and anorectal cancers]. AB - The authors report their 12 years of experience of intra arterial chemotherapy in pelvic recurrences and inoperable advanced stages of uterine carcinoma, rectal cancer and anal cancer. In squamous cell cancers the drug associations were mitomycin C, bleomycin, fluorouracil and folinic acid and cisplatin. In adenocarcinoma the same protocol contained no bleomycin. Drugs were infused for a 48 hours period in continuous infusion. The dosages were the same than in the intravenous regimens. Twenty patients with pelvic recurrences were included in this retrospective study: six were uterine cancers, fourteen were colo rectal cancers and two had advanced stage uterine cancer. Pain decreased in 10/14 patients with ano-rectal cancer pre sacral recurrence. Partial response was observed in 12 patients. Complete secondary surgical resection was possible in 4/14 rectal cancers and 6/6 uterine cancer recurrences. Chemotherapy induced a pathological complete response in 4/6 uterine cervix carcinoma recurrences. These observations led to perform pelvic intra arterial chemotherapy as first line treatment of locally-advanced inoperable pelvic tumors: 11 uterine cancers and five ano-rectal cancers. The objective were: tumor reduction before radiotherapy or surgery, tumor sterilization, decrease tumor volume for better radiation dosimetry, increase the chance of organ-preservation. The observation of tumor reduction in this small number of patients does not allow to draw definite conclusions. However the introduction of intra arterial pelvic chemotherapy as first line treatment of inoperable pelvic cancer warrants further studies. PMID- 10846532 TI - [Methotrexate pneumonitis arising during the treatment of non-Hodgkin's lymphomas with the m-BACOD protocol]. AB - We retrospectively studied 32 patients treated with the m-BACOD regimen in a single institution between January 1988 and December 1991. After four to seven courses, four patients presented severe acute pneumonitis (PaO2 < 55 mmHg in room air), with diffuse bilateral interstitial syndrome. Broncho-alveolar lavage displayed increased lymphocyte count (> 45%) with inversion of CD4/CD8 in two cases and no evidence of parasitic, bacterial or viral infection. All patients received methyl-prednisolone (0.5 to 1 mg/kg/d x 1 week) with both complete clinical and radiological recovery within a week. The m-BACOD regimen was continued without bleomycine for four patients and without bleomycine plus methotrexate for two patients, until the completion of eight courses, without recurrence of pneumonitis. Drug-exclusion decisions were made empirically because the exact nature of the pneumonitis was not recognized at the time of diagnostic. Because of the regular administration in the m-BACOD regimen, methotrexate leads to an increased risk of pneumonitis. We concluded that the use of the m-BACOD regimen should henceforth be discontinued. PMID- 10846533 TI - [Carcinomas of the nasopharynx in children. Analysis of 50 cases]. AB - From 1980 to 1990, 50 children were treated for nasopharyngeal carcinoma in the Department of Oncology at the hospital Center Ibn Rochd in Casablanca. There were 34 boys and 16 girls with a median age of 12.3. UCNT were observed in 70% of cases. Fourty six percent were stage T3-T4 and 70% were N2-N3 disease. All children received irradiation therapy and 16 of them have had a neo-adjuvant chemotherapy. Thirty seven children were evaluable, the immediate response was complete in 26/37 cases. Recurrence of disease was noted for four children and metastatic for four others children. Two children have both recurrence and metastatic disease. The overall survival was 40.5% at 3 years. Disease free survival was 37.8% at 3 years. PMID- 10846534 TI - Surveillance-only policy in clinical stage-I non-seminomatous germ-cell tumors of the testis. AB - With the advanced imaging techniques, the sensitive assays for tumor markers and the curability of small-volume metastatic disease with cis-platinum-based chemotherapy, surveillance has gained popularity in clinical stage-I non seminomatous germ-cell tumors. This study reports our experience on 58 patients who have been included in a surveillance protocol. Patients with normal tumor marker levels following surgery, no evidence of metastases on the CT scans of the abdomen and the chest and no residual tumor at the surgical margin were followed. Relapsing patients were treated with cis-platinum-based combination chemotherapy. Of the 58 patients, 17 relapsed (29.3%) in a period of 2-18 months (median 5 months). None of the relapsing patients later presented with evidence of disease in the follow-up period 14-79 months (median 39 months) after chemotherapy. Prognostic factors were evaluated by univariate analysis and the data for risk factors, such as the presence of embryonal carcinoma, absence of yolk-sac elements and scrotal violation, were evaluated by multivariate analysis. Statistical analysis showed that none of the risk factors were significantly different in predicting the relapse. Of the 6 relapsing patients with preorchiectomy elevated tumor-marker levels, 4 had shown a slower decline in tumor-marker levels than expected and they all relapsed with elevated levels of the same tumor markers. A surveillance-only policy actually seems to be a safe and logical approach if the patients are properly selected and cooperate fully. PMID- 10846535 TI - [Adenocarcinoma of rete testis. Apropos of a case and review of the literature]. AB - We report a case of localized adenocarcinoma of rete testis in a 55 year-old man. Treatment associated radical orchidectomy and retroperitoneal lymph node dissection. At last follow up, the patient was well, without any evidence of recurrence, 26 months after surgery. The literature and the treatment modalities are reviewed. PMID- 10846536 TI - [Taxoids: structural and experimental properties]. AB - Paclitaxel (Taxol) and docetaxel (Taxotere) are the first representatives of a new class of antitumor compounds. These two taxoids are clinically active against breast, ovarian and lung cancers. Taxoids are highly complex diterpenoids form natural origin. Preclinical and clinical developments have been made possible after a long and sustained chemical effort: paclitaxel is extracted from the barks of the Pacific yew tree Taxus brevifolia whereas docetaxel is prepared by hemisynthesis starting from 10-deacetyl-baccatin III, an inactive precursor found in the needles of the European yew tree Taxus baccata. These two drugs are active in various in vitro and in vivo preclinical models (cell lines, cloning of human tumor stem cells, murine grafted tumors, human xenografts). Taxoids constitute a new class of antimitotic agents different from vinca-alkaloids: on the one hand, paclitaxel and docetaxel can be considered as inhibitors of the reaction of depolymerization of microtubules into tubulin; on the other hand, vinca-alkaloids inhibit reaction of polymerization of tubulin into microtubules. An active program of medicine chemistry is done in various pharmaceutical and academic Institutions with two objectives: knowledge of structure-activity relationships and selection of new candidates for clinical trials. PMID- 10846537 TI - [Pharmacology of camptothecin and its derivatives]. AB - 20 (S) Camptothecin was discovered in the early 60's as a result of the intensive screening of natural products by the NCI. Camptothecin lactone was poorly water soluble and was administered as the sodium salt in phase I trials. Despite some encouraging responses in early studies, continued evaluation of this compound revealed severe and unpredictable toxicities such as haemorrhagic cystitis and diarrhoea. The reversible opening of the lactone ring of a camptothecin is pH dependent and yields a ring-opened carboxylate form which has greatly reduced activity in vivo and in vitro. Under physiological conditions, the carboxylate form predominates, but the exact position of this equilibrium in vivo also depends on other factors such as protein-binding and differential metabolism and elimination. The site of action of camptothecin is a complex formed by the nuclear enzyme topoisomerase I and DNA, which represents a novel target for cancer chemotherapy. The principal role of topoisomerase I is the relaxation of DNA required for transcription and replication. The transient covalent complexes formed by the linking of the enzyme and the 3' extremity of a nicked DNA strand are stabilised in the presence of camptothecin and involved in collisions with replication forks. The ensuing arrest of the fork is accompanied by the generation of permanent double-strand breaks which are thought to be responsible for the antiproliferative properties of camptothecin. The acquisition of resistance to camptothecin in cell culture appears, in general, to be due to a reduction in content and activity of topo-isomerase I. Single-point mutations of the gene of the enzyme have been detected in a number of these resistant variants. Camptothecin appears to be a poor substrate of P-glycoprotein and its intracellular accumulation is not appreciably reduced in cells expressing the multidrug-resistant phenotype. Several water soluble and active derivatives of camptothecin have been synthetized of which CPT-11 and topotecan are the most advanced in clinical trials. These compounds represent two different approaches to the problem of the poor water solubility of camptothecin lactone. CPT-11 is a soluble prodrug which is converted in vivo to the highly active SN-38, whereas topotecan itself is water-soluble due to the presence of a tertiary amine substitution which is charged at physiological pH. These two compounds present different pharmacological properties in the clinical setting. PMID- 10846538 TI - [Education of cancer patients]. AB - Increasing patients' information is currently evolving toward real education, mainly concerning chronic diseases. Cancer patients could also benefit from this evolution of medical care. Rationale for such education certainly includes theoretical endpoints (patient's autonomy) but also practical considerations that will improve treatment quality (compliance), tolerance and further rehabilitation. Many patients may benefit from this education during complex and long-term treatments. Furthermore, sequelae and handicaps will be either reduced or better tolerated. This education is provided by practitioners and other care providers. That implies improved specific training and further need for written and video-taped documents. Such an evolution should further lead to proper evaluation and cost-benefit analyses in order to ensure real improvement of both treatment quality and patient's experience. PMID- 10846539 TI - [Autocrine stimulation of cell proliferation in colonic carcinoma: a biological curiosity or a new therapeutic target?]. AB - Chemotherapy has little efficacy in the treatment of advanced colorectal carcinoma. Biological investigation has made evident several autocrine stimulation loops; the best documented one involves epidermal growth factor (EGF): this growth factor stimulates cell proliferation and cell secretion of proteolytic enzymes. Suramin and somatostatin are able to disrupt these loops of stimulation. Clinical studies performed with octreotide, a somatostatin analogue, and suramin have been unsuccessful until now. PMID- 10846540 TI - [Treatment of metastatic kidney neoplasms with a new interleukin 2 protocol: The experience of the Gustave-Roussy Institute]. AB - Treatment of metastatic renal cell carcinoma with interleukin 2 (IL2) remains controversial despite the authorization from the French government for IL2 with the West schedule in this disease. We report herein the study of the Institute Gustave-Roussy of 73 patients, who received from 1989 to 1991 a new schedule of high dose IL2. Seventy three patients received high dose IL2 according to the following schedule: IL2 by continuous infusion at 24.10(6) IU/m2/d, on 2 consecutive days per week, during 5 weeks. This treatment was associated in the first 33 patients with gamma interferon at a dose of 5.10(6) IU/m2/d subcutaneously the days of IL2 infusion, during the 5 weeks of therapy. Immunotherapy was further continued in responding patients, either as an association of IL2 and LANAK (lymphokine-activated natural killer) cells, or as IL2 alone. Finally, when possible, surgery was performed on residual masses. Twenty five percent of objective responses (PR + CR) have been observed. Moreover, 12.3% CR has been obtained after the overall therapy. The global mean survival is 15 months, with a mean survival of 8, 18 and 24+ months depending on the status of the disease (progressive, stable or responding) after initial treatment with IL2. Tolerance of this schedule was good with an actual received dose of 90% of the planned doses, and patients could leave the hospital within 2 hours after the end of IL2 in 87% of the cycles. No toxic death was observed. Among the parameters observed for correlation with the clinical response, only performance status and level of sTNF-alpha R were significantly associated with the response. PMID- 10846541 TI - [Clinical importance of the determination of the hydrophobic forms of serum carcinoembryonic antigen (CEA) in metastatic cancer of the colon]. AB - Thirty-four patients with metastatic colon cancer were treated with 5 fluorouracil and folinic acid. The follow-up of disease was evaluated by tomodensitometric CT-scan analysis and by serum CEA determination. In addition, a study of the different CEA molecular forms separated by Triton X114 partitioning, immunoprecipitation and immunoblotting was completed. Concerning the aqueous phase, no relationship appeared between the pattern of CEA species and the outcome of chemotherapy. Opposingly, the analysis of the hydrophobic phase gave results closely correlated to chemotherapeutic response. In 19/34 patients, the hydrophobic CEA forms were absent or weakly expressed; out of these patients, 16/19 underwent a successful response to chemotherapy regimen. Opposingly, all of the remaining 15 patients expressing high levels of hydrophobic CEA species were non-responders. The present study thus gives new means for predicting the outcome of 5 fluorouracil-folinic acid chemotherapy by screening the molecular CEA forms expressed in the serum of patients with metastatic colon cancer. PMID- 10846542 TI - [Does the professional image of the social worker in oncology correspond to the current challenges of the job? Apropos of a survey of patients and care professionals]. AB - Care professionals (physicians and nurses), administrative staff and hospitalised patients were interviewed to find out the necessary qualities for social workers in oncology, as well as the roles and tasks that they must fulfil. On the one hand, the results of this study show that the needs of the interested parties are indeed met by social service from an empirical perspective of psychosocial support and expert advice. On the other hand, they bring to light the fact that the current status of the social worker is hardly recognized. Confronted with new occupational challenges, the social worker should rethink his/her profession and reevaluate his/her practice. Furthermore, he/she should develop professional links intra- and extramural and participate in social research programs whose aim is to improve the rehabilitation of patients and their families. PMID- 10846543 TI - Functional nerve growth factor receptor in von Recklinghausen neurofibromatosis: an immunocytochemical and short-term culture study. AB - Immunocytochemistry reveals 75 kDa low affinity type nerve growth factor receptor (NGFR) on the cell membrane of human neurofibroma cells of von Recklinghausen disease in vivo and in vitro. NGF-immunoreactivity is detected in the primary and cultured tumor cells. Growth augmentation of cultured neurofibroma cells by exogenous NGF is also confirmed. Phosphotyrosine-immunoreactivity is demonstrated by immunocytochemistry in the in vivo and in vitro neurofibroma cells suggesting possible phosphorylation of tyrosine residue in the NGFR or a cellular protein downstream of signal transduction through the ligand receptor system. These results indicate human neurofibroma cells possess functional NGFR and the growth is potentiated through the NGF-NGFR system in the paracrine and/or autocrine fashion. PMID- 10846544 TI - Expression of bone morphogenetic proteins in human osteogenic and epithelial tumor cells. AB - In addition to osteosarcomas some epithelial tumors also show bone induction, which is supposedly due to bone morphogenetic proteins (BMPs). However, the types of BMPs expressed in different kinds of tumors have not been studied extensively. In this study five human osteosarcoma and six human carcinoma cell lines were examined to investigate the bone induction and BMP mRNA expression. The cultured cells were transplanted in the subcutaneous tissue of athymic nude mice with or without diffusion chambers. Reverse transcription-polymerase chain reaction (RT PCR) was used for the detection of BMP mRNAs. All osteosarcoma cell lines expressing all BMPs (BMP-1 to 7) examined, successfully formed bone or osteoid tissue irrespective of the use of a diffusion chamber. Bone formation was induced by diffusion chamber in an osteosarcoma cell line OST and a gastric adenocarcinoma cell line MKN45. OST cells lacked BMP-3 and 7, and MKN45 cells lacked BMP-3, 5 and 6. Our data suggest that BMP-3, 5, 6 and 7 are not always essential for bone induction in tumor cells. PMID- 10846545 TI - The immunophenotype of perivascular cells in the human brain. AB - The immunophenotype of perivascular cells (PC) in temporal lobe tissues obtained at autopsy in 48 patients (aged 41-88 years) was characterized using light and electron microscopic immunocytochemistry with a variety of antibodies. In all cases studied, PC bearing CD11c (Ki-M1P) and CD68 (KP1) were distributed throughout the temporal cortex. In addition to Ki-M1P and KP1, the monoclonal antibodies against major histocompatibility complex (MHC) class II antigen (Ag) (LN-3, CR3.43), anti-leucocyte common antigen (LCA), LN-5, Mac 387 were all found in PC with variable immunoreactivity. In contrast, LN-1 and OPD4 were not found in PC, although the former showed nearly constant staining of resting microglia. Semiquantitative analysis disclosed differences in the numbers of cells labeled with the markers in the 21 normal brains (Ki-M1P > KP1 >> LCA, LN-3, LN-5 >> Mac 387). Ultrastructurally, immunoreactivity for Ki-M1P, KP1, and LN-3 was observed in PC with cytoplasm containing dense lysosomal bodies. In brains from patients with Alzheimer's type dementia, PC were seen in the wall of beta-amyloid protein positive small vessels. However, there was no definite alteration of antigenicity in PC from AD brains compared with those from normal brains. The immunophenotype of PC was similar to that of macrophages, which were observed in the perivascular spaces and the leptomeninges in some normal and diseased brains. In contrast with PC, however, macrophages showed high incidence of labeling for some macrophage markers LN-5 and Mac 387. These findings demonstrate that PC may be a normal constituent of the adult human brain with a variable expression of monocyte/macrophages markers and MHC class II Ag and that PC could be distinguished from resting microglia by their morphology and by their immunophenotype. PMID- 10846546 TI - Immunohistochemical study of hepatic angiomyolipoma. AB - An immunohistochemical study was performed on nine hepatic angiomyolipomas (AML) found in eight patients. Histologically, the tumors were fundamentally composed of the three heterogeneous tissue components of blood vessels, smooth muscle cells (SMC), and fat cells, although the proportions and distributions were quite variable from tumor to tumor and from area to area in the same tumor. Additionally, cellular pleomorphism and atypia with occasional bizarre giant cells were found in the SMC component. This histologic feature might lead to a mistaken diagnosis of malignant neoplasm, and pathologists should therefore be aware of the broad histologic spectrum of hepatic AML. However, the immunostaining patterns were basically the same in all nine tumors. All tumor components were negative for epithelial membrane antigen (EMA) and for cytokeratin. The spindle-shaped SMC component of the tumor was occasionally positive for vimentin, desmin and alpha-smooth muscle actin, whereas epithelioid SMC were negative for all three. Both the epithelioid and spindle-shaped SMC were occasionally positive for S-100 and neuron-specific enolase. All types of SMC in the tumor, whether spindle, epithelioid, intermediate or pleomorphic SMC, were strongly positive for HMB-45, a melanoma-specific monoclonal antibody. Fat cells were occasionally positive for S-100. Endothelial cells were positive for factor VIII-associated antigen. Among hepatic tumors HMB-45 reactivity is, so far as we know, found exclusively in the SMC of AML, and the HMB-45 reactivity of a hepatic tumor is thus clearly an important piece of information in the diagnosis of AML. PMID- 10846547 TI - Apoptosis of colon cancers assessed by in situ DNA nick end-labeling method. AB - Apoptosis in colon cancers was investigated using in situ DNA nick end-labeling. Seventy-six colon cancer cases were divided according to histologically defined malignant grading, degree of invasion and other pathologic indicators. The apoptotic labeling index (ALI) was highest in cases with invasion limited to the submucosa, the difference as compared to other stages being statistically significant, while no correlation was noted between ALI and histological type. ALI significantly decreased with lymph node involvement of cancers. In contrast, there appeared to be no link with vessel invasion. ALI was significantly higher in lesions smaller than 2 cm in diameter. These results indicate that apoptosis in colon cancers is more frequent at a relatively early stage when the lesions are small, and suggest that the apoptotic phenomenon may play some role in regulating the size and progression of such tumors. PMID- 10846548 TI - Alteration of the p53 gene of lung carcinomas with sarcomatous transformation (spindle cell carcinoma): analysis of four cases. AB - Lung carcinoma with sarcomatous transformation (LCST) is highly aggressive and characterized by local invasion and/or distant metastasis, which leads to a shorter survival than ordinary lung carcinomas. Therefore, to elucidate whether the malignant potential of the spindle cell element in LCST is associated with the alteration of the p53 gene, four cases were examined by analyses of overexpression of the p53 oncoprotein, mutation of the p53 gene and loss-of heterozygosity (LOH) at chromosome 17p. In two cases overexpression of the p53 oncoprotein of the spindle cell component showed a higher degree of staining than that of the carcinoma component; LOH was identified in both carcinoma and sarcomatous components in one case, while in contrast, another case showed LOH in the sarcomatous component only. Mutations were clearly detected in two cases; one showed a CTT to CGT transversion in codon 194 of exon 6 in both components, whereas the other showed a CTG to CAG transversion in codon 265 of exon 8 in the sarcomatous component only. On the basis of these observations, it suggested that the sarcomatous component shows a higher frequency of p53 gene abnormalities in comparison to the carcinoma component. These results also suggested that the acquisition of malignant potential in the sarcomatous component, or the morphological alteration of carcinoma cells, is correlated with abnormalities associated with the p53 gene. PMID- 10846549 TI - Overexpression of p53, c-erbB-2 and epidermal growth factor receptor in human breast carcinomas. AB - Overexpression of p53 protein, epidermal growth factor receptor (EGF-R), and c erbB-2 protein was assessed by immunohistochemical staining of formalin-fixed, paraffin-embedded tissue from 64 invasive breast tumors. The correlation between abnormal expression of each protein and various disease parameters, including lymph node metastasis and histopathologic type and grade was analyzed. Despite the previous proposal, no significant correlation was found between lymph node metastases and overexpression of each gene in the primary tumors. In addition, some metastatic lesions did not always exhibit overexpression, even if it was evident in the primary tumors. Overexpression of c-erbB-2 protein correlated well with Bloom's histological grading. p53 expression was detected most often in tumors with hyperchromatism and more frequent mitosis. Overexpression of c-erbB-2 protein occurred more frequently in p53-positive tumors. The results indicate that abnormal expression of p53 protein causes genetic instability in the early stage of tumor development, resulting in subsequent overexpression of other oncogenes. PMID- 10846550 TI - Biphasic intra-abdominal desmoplastic small cell tumor in a patient with proximal spinal muscular atrophy. AB - A case is reported of intra-abdominal desmoplastic small cell tumor (IDSCT) with biphasic histologic features in a patient with proximal spinal muscular atrophy. The tumor was composed of small epithelial cell nests with spindle cell sarcomatous areas. Both areas were surrounded by a desmoplastic stroma. Immunohistochemical studies revealed reactivity for low molecular weight cytokeratin, epithelial membrane antigen, vimentin, desmin and Leu-7 in both areas. Electron microscopic examination demonstrated paranuclear aggregates of intermediate filaments, zonula adherens and basement membrane-like material in the epithelial cells, while spindle cells in the tumor had fewer intracytoplasmic organelles. However, intermediate or transitional forms of both types of tumor cells were frequently observed. Although IDSCT are known to express multi phenotypes immunohistochemically, attention should be paid to the broad spectrum of cell morphology in these tumors. PMID- 10846551 TI - Mammary hamartoma: immunohistochemical study of two adenolipomas and one variant with cartilage, smooth muscle and myoepithelial proliferation. AB - Three cases of mammary hamartoma were investigated immunohistochemically and are described. Case 1 was a 42 year old woman with an elastic hard tumor, 1.5 cm in diameter, in her left breast. Case 2 was a 49 year old woman with a semisoft tumor, 5 x 2 cm, in her right breast. Case 3 was a 47 year old woman with a hard tumor, 5 cm in diameter, in her left breast. In each case, mammography and ultrasonography revealed a benign-looking, well-circumscribed mass without calcification. Histologically, the tumors were composed of adipose tissue, mammary glands, and fibrous and/or fibromuscular tissue. The tumor in case 3 also contained small islands of hyaline cartilage. Immunohistochemical analysis was performed, and epithelial and mesenchymal components were discretely and differentially immunostained except that the smooth muscle component seemed to be derived from myoepithelial cells. Cartilage formation might be the result of metaplasia, and 'metaplastic variant of the mammary hamartoma' or 'choristoma' may be an appropriate term for cartilage-containing mammary hamartoma. Using proliferating cell nuclear antigen (PCNA)-immunostaining, we observed that each component of the tumors had an individual growth rate. This finding may reflect one aspect of the biological characteristics of hamartoma. PMID- 10846552 TI - Nodular pulmonary amyloidosis associated with asbestos exposure. AB - A 71 year old man was admitted for the purpose of diagnosis of a right solitary pulmonary nodule. The size of the nodule was 18 x 18 mm in diameter 2 years ago, but it has become large, 25 x 25 mm in diameter. The nodule was resected by thoracotomy. Microscopically, eosinophilic amorphous, acellular substances were surrounded by inflammatory infiltrates. It stained with Congo red stain and showed green birefringence with polarizing microscopy. Amyloid fibrils were observed electron microscopically. Asbestos bodies were observed in the lung parenchyma around the nodule. This case shows that a nodule in nodular pulmonary amyloidosis can grow gradually and suggests the possibility of asbestos fibers as one of the etiologic factors in nodular pulmonary amyloidosis. PMID- 10846553 TI - A case of malignant transformation in thoracic vertebral hemangioma following repetitive irradiation and extraction. AB - We report a rare case of thoracic vertebral hemangioma which developed into angiosarcoma during the course of repetitive operations and irradiation. A 44 year old female was operated on for hemangioma of the first thoracic vertebra. The diagnosis of hemangioma was confirmed histopathologically with the specimen from the first operation. The tumor developed multiple lesions later in the clinical course after the first operation, these lesions were removed in four consecutive operations and each histological diagnosis was that of hemangioma. Throughout the period of these operations, the patient was treated with steroid, and with radiotherapy simultaneously. The patient underwent the fifth operation for the recurrence of the tumor on 26 March 1990, and the histopathological diagnosis was not hemangioma but hemangiosarcoma which was considered a malignant transformation. The tumor cells immunohistochemically revealed positive staining with UEA-I, Factor-VIII, as the tumor immunohistochemically showed a vascular endothelioid nature. PMID- 10846554 TI - Ossifying fibromyxoid tumor of the retroperitoneum. AB - A case of ossifying fibromyxoid tumor (OFMT) in an unusual site, the retroperitoneum, in a 71 year old Japanese man is reported. The well demarcated tumor contained a large amount of mature bony tissue within the tumor. The tumor cells were round to spindle-shaped, and showed a high cellularity in some areas and were loosely arranged in other areas. Although the majority of tumor cells expressed vimentin, neuron specific enolase and Leu-7, there were no tumor cells positive for S-100 protein, myogenic or epithelial markers. We would like to emphasize that tumors classified as OFMT contain a wide spectrum of histologic features and heterogeneity. PMID- 10846555 TI - Fibrolamellar hepatocellular carcinoma: a report of a resected case with an electron microscopic and flow cytometric analysis [corrected]. AB - A resected case of fibrolamellar (FLC) and hepatocellular (HCC) combined carcinoma arising in a non-cirrhotic liver of a 29 year old female is reported, including results of the preoperative percutaneous aspiration biopsy, which suggested FLC, and postoperative electron microscopic and flow cytometric analysis. Sections of the resected massive tumor of the left lobe of the liver showed hepatocellular carcinoma accompanying the fibrolamellar carcinoma element which was composed of tumor cells with eosinophilic granular cytoplasm and unique cytoplasmic vacuoles (pale bodies). Lamellar fibrosis was present in the stroma, while no macroscopic central scar was demonstrated. Electron microscopy showed typical features of FLC and flow cytometric DNA analysis indicated diploid DNA pattern with a low proliferation rate. A common HCC element with trabecular structure also existed at the periphery of the tumor. No apparent etiologic agent for the development of hepatic neoplasm was indicated in the history of this patient. She had been without recurrence for about 3 years after extended left lobectomy, when local recurrence was revealed. The recurrence has been treated with chemoembolization and percutaneous ethanol infusion several times up till the present. This case reconfirms the importance of the pathological diagnosis of FLC to promote surgical intervention. PMID- 10846556 TI - Co-expression of TNF alpha and IL-1 beta in human acute pulmonary fibrotic diseases: an immunohistochemical analysis. AB - To clarify the involvement of TNF alpha and IL-1 beta in the initiation of fibrotic lung diseases, their localization was examined by immunohistochemistry. Fibrotic lung diseases observed were classified into acute and old pulmonary fibrotic changes. The acute fibrotic changes included adult respiratory distress syndrome, acute interstitial pneumonia and idiopathic pulmonary fibrosis with acute exacerbation. Acute pulmonary fibrotic changes histopathologically corresponded to a mixture of the exudative and proliferative phases of diffuse alveolar damage. Both TNF alpha and IL-1 beta were positive in the alveolar macrophages and proliferating type II pneumocytes in acute fibrotic changes. In contrast, positive cells for TNF alpha and IL-1 beta were sparse in the areas of old fibrotic change and in the normal tissue. These findings suggest that TNF alpha and IL-1 beta play an important role in the initiation of pulmonary fibrotic responses and in the architectural remodeling, irrespective of the etiology of fibrotic lung diseases. PMID- 10846557 TI - Basement membrane thickening in the placentae from diabetic women. AB - A light microscopy study was carried out on 48 placentae. Seventeen placentae were obtained from non-diabetic mothers while the other 31 placentae were from both women with controlled diabetes and women who had an abnormality of the glucose tolerance test. All the women delivered at 38-40 weeks of gestation. Placentae from diabetic patients showed immaturity of the villi, hypertrophy of the capillaries and thickening of the basement membrane of the trophoblastic villi (3.2 +/- 0.35 microns) and the amniotic membrane (1.8 +/- 0.3 microns). Focal fibrinoid necrosis, an increase in the number of Hofbauer cells and dilatation of villi capillaries were also commonly observed in placentae from diabetic mothers, and the normal cuboidal cells lining the amniotic membrane tended to become tall columnar (17.6 +/- 6.3 microns) with distally located nuclei. Similar findings were observed in patients who had a potentially abnormal glucose tolerance test, which suggests the possibility of primary lesion in origin. Therefore, control of hyperglycemia may only partially prevent the development of placental abnormalities. PMID- 10846558 TI - Cell renewal and functional morphology of human lactating breast: presentation of a new mode of cell death ('magentosis') characterized by nuclear periodic acid Schiff reactivity. AB - Cell kinetics and functional morphology of the human lactating breast were analyzed using 21 paraffin-embedded specimens of the lactating breast obtained by biopsy or surgery and two of the 'resting' breast. Four types (I-IV) of lactating lobules were categorized histologically, and were well correlated with their functional status demonstrated by immunohistochemistry for epithelial membrane antigen, beta-casein, lipase, lactoferrin, secretory component and IgA. Type I corresponded to a pre-lactating state, type II to an actively lactating state, type III to an early stage of regression, and type IV to an advanced stage of regression. Cell proliferation monitored by Ki-67 (MIB-1) immunostaining was at the highest level in type I lobules followed by type IV, while the labeling indices were low in types II and III. Apoptosis demonstrated by terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate (dUTP) biotin nick end-labeling (TUNEL) method showed labeling profiles comparable with the proliferative activity. Holocrine-type desquamative cells with intact appearing nuclei were frequently recognized in type II lobules. In type III, a peculiar form of cell death, designated 'magentosis', was demonstrated. These degenerative cells occasionally shedding into the acinar lumen possessed homogeneous pale, TUNEL-negative nuclei with diastase-resistant periodic acid Schiff (PAS) reactivity. 'Magentosis' specifically identified in an early phase of involution of the human lactating breast might represent a unique mode of cell death distinguishable from apoptosis and necrosis. PMID- 10846559 TI - 'Magentosis' in human lactating breast: a mode of cell death accumulating single stranded DNA stretches or breaks. AB - Pale and homogeneous-looking nuclei of degenerative acinar cells selectively seen in an early regression stage of the human lactating breast were periodic acid Schiff (PAS)-reactive. In our preceding paper, this peculiar morphologic feature was designated as 'magentosis'. The present paper was aimed at histochemically clarifying the nature of the 'magentotic' nuclei. The diffuse PAS reactivity was not influenced by pretreatments with alpha-amylase, DNase, RNase, proteinase K, nor by hydrochloric acid or heating. The nuclei were negative for acid mucosubstances and secretory glycoproteins, and were unreactive with a variety of lectins. In contrast, the presence of single-stranded DNA stretches or breaks was proven. The 'magentotic' nuclei in non-heated paraffin sections were hybridized with a heat-denatured DNA probe for human DNA consensus sequences and were focally immunoreactive with an antibody to single-stranded DNA. The terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) method turned to be positive after digestion by mung bean nuclease, a single-stranded DNA-specific enzyme. The 'magentotic' nuclei were further clearly labeled by the in situ nick translation method. The nucleoli were devoid of reactivity for both the PAS and single-stranded DNA signals. We propose that 'magentosis' represents a unique mode of cell death, distinct from apoptosis and necrosis or oncosis. The PAS reactivity in the 'magentotic' nuclei may be correlated with the occurrence of single-stranded stretches or breaks in the DNA chain. PMID- 10846560 TI - The relationship between the gene mutation of p53 and the protein expression of p53 and Ki-67 in non-Hodgkin's lymphomas. AB - The relationship between the mutation of the p53 gene and the expression of the p53 protein and the Ki-67 antigen has been investigated in 115 cases with non Hodgkin's lymphoma, using the immunohistochemical double staining technique, single-strand conformational polymorphism and DNA sequencing methods. Eighteen cases showed more than 10% of p53+ cells and the others showed a few p53+ cells presented sporadically. Alterations in the p53 gene were detected in six cases with B cell type, consisting of five cases with point mutation and one case with point mutation and 15 base pairs deletion. These six cases showed a high percentage of p53+ cells and five cases revealed that the percentage of p53+ cells was higher than that of Ki-67+ cells (p53+ cells > Ki-67+ cells). Excluding the six cases with mutation of the p53 gene, all cases revealed that the percentage of p53+ cells was less than that of Ki-67+ cells (p53+ cells < Ki-67+ cells). Moreover, there was a positive correlation between expression of the p53 protein and of the Ki-67 antigen in histologic types of B cell lymphomas and of T cell lymphomas, respectively, except in small non-cleaved (Burkitt's) and lymphoblastic types. Therefore, sporadic cases showing p53+ cells > Ki-67+ cells revealed alteration of the p53 gene, and expressed abnormal p53 protein (mutant form). Most cases showing p53+ cells < Ki-67+ cells expressed normal p53 protein (wild type), and may reflect the rapid proliferation rate. PMID- 10846561 TI - Proliferating cell nuclear antigen expression in Wilms' tumor: a comparative study of pre- and post-chemotherapy specimens. AB - A total of 36 Wilms' tumors were immunohistochemically analyzed for proliferative (proliferating cell nuclear antigen-labeling) index (PI) with special reference to the effect of preoperative chemotherapy on the proliferative potential of tumors and differences in PI according to the histologic components (epithelial, blastemal and stromal). The cases were divided into two groups according to the status of chemotherapy at the time of surgical removal of the tumors. In group I (n = 12, cases with pre-operative chemotherapy), the mean PI +/- s.d. values of epithelial, blastemal and stromal types were 19.5 (n = 1), 10.1 +/- 5.28 (n = 8), and 16.2 +/- 13.49 (n = 3), respectively, and in group II (n = 24, cases without preoperative chemotherapy), 45.2 +/- 15.41 (n = 3), 28.5 +/- 16.92 (n = 10), and 23.2 +/- 4.45 (n = 2), respectively. The cases with mixed histology in group II (n = 9) had comparable PI values for each histologic component. The mean PI +/- s.d. was 12.1 +/- 7.76 in group I and 31.9 +/- 15.95 in group II. The PI value of group II was significantly higher than that of group I (P < 0.001). Alterations in the PI value following chemotherapy implied that the interpretation of tumor histology following chemotherapy should be more prudent because alteration of the PI, which reflects the proliferative potential of tumor cells, suggests modified biology of viable cells in pretreated cases as a consequence of chemotherapy. PMID- 10846562 TI - Correlation of two argyrophilic nucleolar organizer region counting methods with the Ki-67 labeling index in uterine smooth muscle cell tumors. AB - The argyrophilic nucleolar organizer regions (AgNOR) are silver stained granules that are thought to correlate with cell proliferation activity. Two AgNOR counting methods: the mean AgNOR count (mAgNOR, the mean number of AgNOR granules in 100 cells) and the AgNOR proliferative index (pAgNOR, the percentage of cells exhibiting five or more AgNOR granules per nuclei) have been proposed. In this study, the two counting methods were applied to 58 cases of normal uterine corpus and uterine corpus tumors and were compared with the Ki-67 labeling index (Ki-67 LI) using MIB-1 monoclonal antibody and other histopathological criteria. Notable differences in the number of AgNOR and the Ki-67 LI were observed between benign and malignant smooth muscle tissue. Histopathologic features are well correlated to the proliferative activity of tumors. Although the most reliable method of predicting malignant potential cannot be determined, the methods outlined by this study are thought to be highly useful in assessing proliferative activities. PMID- 10846563 TI - Expression of classical protein kinase C subspecies in non-neoplastic lymphocytes and non-Hodgkin's lymphomas: an immunohistochemical study. AB - It is generally accepted that phosphorylation plays a pivotal role in the cellular response of cell differentiation and proliferation. Immunohistochemical expression of classical protein kinase C (cPKC) subspecies (alpha, beta and gamma) in eight reactive lymphoid tissues, three normal spleens and 149 non Hodgkin's lymphomas was examined. cPKC beta was observed primarily in the mantle zone B cells, but appeared as very faint staining in Ki-67 positive proliferated B cells in the germinal centers of secondary lymph follicles. In contrast to the reactive state, high levels of cPKC subspecies were recognized in the majority of 149 cases of non-Hodgkin's lymphoma, including those thought to have arisen from germinal center cells such as follicular lymphoma. The expression of cPKC alpha was found in higher frequency in T cell lymphomas than B cell lymphomas (P < 0.01) by the Chi-squared test. High levels of cPKC alpha were present only in high grade or highly aggressive lymphomas, showing the highest incidence in the small non-cleaved cell type, according to the International Working Formulation and National Cancer Institute (P < 0.01). cPKC gamma was not detected in normal lymphoid cells and was expressed in only four cases of non-Hodgkin's lymphomas. It is presumed that cPKC alpha and beta have a relationship to cell activation and proliferation of lymphoid cells of reactive and neoplastic states. It might be considered that the expression of cPKC alpha may have a relationship with aggressiveness in non-Hodgkin's lymphomas. PMID- 10846564 TI - Immunophenotypical comparison of Gaucher's and pseudo-Gaucher cells. AB - An immunohistochemical study on bone marrow biopsies and spleens of patients with Gaucher's disease and chronic myeloid leukemia was performed to investigate the immunophenotype of Gaucher's cells and pseudo-Gaucher cells. A panel of antibodies was used which were reactive on paraffin-embedded tissues and directed against different hematopoietic lineage cells. Gaucher's cells and pseudo-Gaucher cells expressed a very similar immunophenotype and displayed an intense reaction for the monocytic antibodies tested, thus confirming their common origin and that they belong to the same system. The expression of HLA-DR antigens was much stronger in Gaucher's than in pseudo-Gaucher cells. This last finding, together with other serological data, suggests that Gaucher's cells could not behave simply as phagocytic cells but might play an active role in the chronic stimulation of the immune system that sometimes occurs in patients with Gaucher's disease. PMID- 10846565 TI - Cervical intra-epithelial neoplasia (CIN) III of the uterine cervix in a young female associated with AIDS. AB - A case of cervical intraepithelial neoplasia (CIN) III implying severe dysplasia or carcinoma in situ of the uterine cervix in a 24 year old Japanese female patient with acquired immunodeficiency syndrome (AIDS) is reported. Autopsy revealed marked systemic atrophy of lymph nodes, Pneumocystis carinii pneumonia, pulmonary aspergillosis, acute pancreatitis and CIN III of the portio vaginalis uteri. The human papillomavirus (HPV) genome was detected in sections of the CIN III by polymerase chain reaction. This is the first autopsy report of cervical neoplasia in a patient with AIDS in Japan, and the pathogenesis of CIN III in this case was associated with HPV infection. Gynecological examination should be mandatory for female patients with human immunodeficiency virus infection. PMID- 10846566 TI - Crystalglobulin-induced vasculopathy accompanying ischemic intestinal lesions of a patient with myeloma. AB - A 37 year old man with multiple myeloma (IgG lambda) developed perforation of the small intestine and had gangrene in his feet. In the resected small intestine and below-knee amputated feet, intravascular deposition of crystals, immunohistochemically positive for IgG and a lambda light chain, were found. Although paraproteins produced in the course of plasma cell dyscrasia may crystallize and cause systemic occlusive vasculopathy, this is the first case with intestinal involvement. Because cryoglobulinemia was absent in this patient, crystals may be derived from the myeloma protein which sometimes crystallizes in the blood without the influence of low temperature. PMID- 10846567 TI - Participation of endothelial cells in the development of glomerulosclerosis: a study on murine serum sickness nephritis with mitomycin C. AB - To investigate the participation of endothelial cells in glomerulosclerosis, the study was performed in serum sickness nephritis (SSN) with administration of mitomycin C (MMC). SSN was induced in 8 week male Fisher rats by sensitizing them with albumin, chicken egg (EA). Then MMC (0.5 mg/kg bodyweight) was injected daily for 3 days and they were killed at 1, 2, 4 and 6 week intervals. Significant mesangial expansion and sclerosis were observed in the experimental mixed SSN-MMC group in comparison to the SSN or MMC control group from 1 week to 6 weeks (P < 0.05). Moreover at 1 week, double contour appearance of the glomerular capillary wall, basement membrane splitting and disruption were observed light microscopically in the mixed SSN-MMC group. Electron microscopy revealed peripheral capillary basement membrane disruption with huge subepithelial, mesangial osmiophilic deposits and epithelial foot process effacement. At 6 weeks, disappearance of the endothelial cell fenestration and subepithelial basement membrane-like material formation were observed in the MMC group. Based on these results, it is suggested that MMC induced assault on the glomerular endothelial cell produces prominent glomerular capillary basement membrane disruption at the early phase of SSN, resulting in the accumulation of huge subepithelial and mesangial deposits, mesangial cell proliferation, production of the extracelluar matrix component and initiation of glomerulosclerosis. PMID- 10846568 TI - Utility of pancreatic digestive enzyme immunohistochemistry in the differential diagnosis of hepatocellular carcinoma, cholangiocarcinoma and metastatic adenocarcinoma of the liver. AB - To test the diagnostic utility of pancreatic digestive enzyme immunohistochemistry in liver cancers, the expression of three pancreatic digestive enzymes (trypsinogen, chymotrypsinogen and pancreatic lipase) was investigated in cholangiocarcinoma (CC) (n = 42), hepatocellular carcinoma (HCC) (n = 35), combined HCC-CC (n = 11) and metastatic adenocarcinoma (MA) of the liver (n = 34; 4 gastric cancer, 5 pancreatic cancer and 25 colon cancer). In CC, 15 (36%) expressed one or more of these enzymes, while the remaining 27 (64%) did not express any enzymes. In MA, 13 (38%) expressed one or more of these enzymes, while the remaining 21 (62%) did not express any enzymes. Expression of trypsinogen, chymotrypsinogen and lipase was noted in 15 CC (36%), 11 CC (25%) and 15 CC (36%), respectively, and in 9 MA (26%), 6 MA (18%) and 13 MA (38%), respectively. There was no significant difference in the positive ratio of each enzyme between CC and MA. In positive cases, the enzymes were expressed with a cytoplasmic granular pattern. In MA, there was no significant difference in the positive ratio of the enzymes among the primary sites. In contrast to CC and MA, these enzymes were not expressed in any cases of HCC and combined HCC-CC. These data suggest that pancreatic digestive enzyme immunohistochemistry may be useful for differential diagnosis between HCC and CC or MA as well as between combined HCC-CC and CC or MA, but it is not useful for differential diagnosis between CC and MA. A positive reaction for these enzymes is indicative of CC or MA and is against the diagnosis of HCC or combined HCC-CC, and a negative reaction is noncontributory to the differential diagnosis. PMID- 10846569 TI - Myocardial ischemia due to vascular systemic amyloidosis: a quantitative analysis of autopsy findings on stenosis of the intramural coronary arteries. AB - A case is reported of a 65 year old man who suffered myocardial ischemia resulting from extensive stenosis of the intramural coronary arteries secondary to systemic vascular involvement by primary amyloidosis. In the myocardium, there were multiple fibrotic foci scattered mainly in the subendocardial region of the ventricle. Intramural coronary arteries were stenotic or occlusive due to amyloid induced luminal narrowing, but there was no significant stenosis of the epicardial coronary arteries. Quantitative analysis of amyloid deposits in the intramural coronary arteries demonstrated that occlusive arteries were predominant in the surrounding area of myocardial fibrosis, and the extent of coronary stenosis by amyloid deposition was significantly more severe than in hearts of the five control patients who had coronary amyloidosis without myocardial fibrosis. These results indicate that myocardial fibrosis originates from coronary ischemia due to vascular amyloid deposition. This is the first time that the relationship between myocardial lesions and coronary amyloid deposition has been elucidated using histopathologic quantitative analysis. PMID- 10846570 TI - Comparison of proliferative activities and metastases between two subtypes classified at the deeply infiltrating sites of colorectal moderately differentiated adenocarcinomas. AB - Twenty-eight moderately differentiated adenocarcinomas invading beyond the muscularis propria of the colorectum were subclassified as 13 moderate- and 15 poor-subtype tumors based on the histology at the deeply infiltrating sites. Moderately differentiated cancer cells were correlated with liver metastasis and p53 immunoreactivity. Poorly differentiated cancer cells were correlated with lymph node metastases but not to p53 immunoreactivity. The proliferative cell nuclear antigen (PCNA) labeling index (LI), Ki-67 LI and agyrophilic nuclear organizer regions (AgNOR) values determined for the poorly differentiated cancer cells in the poor-subtype tumors were significantly lower than those of moderately differentiated cancer cells in the moderate-subtype tumors. In cells from tumors classified as poor-subtype, poor differentiation was associated with decreased PCNA LI levels, but with unchanged Ki-67 LI and AgNOR values. These results indicate that colorectal adenocarcinoma cells that are histologically subclassified as moderately differentiated have different proliferative and metastatic activities from cancer cells that are poorly differentiated. Moderately differentiated cancer cells are associated with hematogenous metastasis to liver and high proliferative activity, and loss of tubular formation of cancer cells may be fundamentally related to lymph node metastasis and infiltrative growth. PMID- 10846571 TI - Argyrophilic nucleolar organizer regions in colorectal malignancy: application of typing by their density. AB - Colorectal neoplasms obtained at colonoscopy were examined by argyrophilic nucleolar organizer region (AgNOR) staining to evaluate the usefulness of AgNOR as a discriminant for malignancy. AgNOR dots were divided into two kinds: 'structures' (larger and less-densely stained) corresponding to the nucleolus, and 'units' (smaller and densely stained) presumed to be true AgNOR within the structure. The number of structures per nucleus did not differ between the adenoma and carcinoma groups, whereas the number of units per nucleus showed a significant difference. However there were several cases showing an overlap between the adenoma and carcinoma groups, leading to a difficulty in deciding whether any given case was benign or malignant. Three types of AgNOR patterns were categorized based on the ratio of units to structure. Type I was defined as the unit being indistinguishable from the structure, Type II as each structure having one to five units, and Type III as at least one structure having six or more units, irrespective of total number of units per nucleus. The colorectal lesions in which more than half of the neoplastic cells showed Type III coincided well with carcinomas histologically diagnosed, with the exception of adenomas with severe atypia. Labeling index of proliferating cell nuclear antigen (PCNA LI) differed between the adenoma and carcinoma groups with a considerable extent of overlap, and correlated to some extent with the AgNOR values. These results showed that the AgNOR staining was useful for determining malignancy and its usefulness appeared superior to PCNA LI. PMID- 10846572 TI - Collagenous colitis in a Japanese patient. AB - A case of collagenous colitis is reported. A 48 year old female who had been complaining of mild diarrhea had been under medication for a gastric ulcer. Colonoscopy revealed almost normal appearance of the colonic mucosa except for one hyperplastic polyp of the cecum. Specimens of the ascending, transverse and descending colon showed a distinctively thickened collagen band beneath the surface epithelium, 10-20 microns thick, which was irregularly distributed, even within the same specimen. In some areas, the thickened collagen band was found around the upper part of the pits. Periodic acid-Schiff (PAS), Azan staining and silver impregnation were positive for this thickened collagen band. Immunohistochemically, the thickened collagen band was weakly positive for collagen type III, but negative for collagen types I and IV. Plasma cells, lymphocytes and eosinophils were observed in the lamina propria in addition to intraepithelial lymphocytic infiltration. Capillaries were increased in the thickened collagen band. The arrangement of surface epithelial cells was irregular. Crypts were not distorted. Edema, diffuse or extensive fibrosis and congestion were not found. Through these findings the patient was diagnosed as having collagenous colitis. Many cases of this had been reported in western countries, but very few in Japan. The Japanese literature was reviewed for cases of collagenous colitis and it was found that only two cases had been presented. PMID- 10846573 TI - Metastasis from small cell carcinoma of the lung producing acute appendicitis. AB - A case of acute gangrenous appendicitis with perforation caused by metastatic small cell carcinoma of the lung in a 65 year old man is reported. The manifestation of appendicitis occurred more than 4 years after the diagnosis of the bronchogenic carcinoma. With longer survival of patients with disseminated tumors it is probable that new manifestations of those malignancies will be discovered. Acute appendicitis due to metastasis from a distant neoplasm should be considered in the differential diagnosis of right lower abdominal pain in the oncology patient. PMID- 10846574 TI - Angiomyolipoma mimicking true lipoma of the liver: report of two cases. AB - Hepatic angiomyolipoma (AML) is very rare and only about 80 cases have been reported. The tumor is fundamentally heterogeneously composed of the three tissue components of blood vessels, smooth muscle cells (SMC), and fat cells. Two cases of hepatic AML are reported here, both of which are histologically composed predominantly of a fat cell element and resembled true lipoma (lipomatous AML). However, careful examination of both tumors revealed the presence of a small amount of epithelioid SMC, especially around blood vessels. Immunohistochemical study using monoclonal antibody for melanoma (HMB-45) clearly revealed a small amount of HMB-45-positive SMC around the blood vessels and scattered in the diffuse fat cell growth in both tumors. Since no liver tissue components or primary liver tumors are reactive with HMB-45 except AML cells, the presence of HMB-45-positive cells within the tumor clearly established the diagnosis of hepatic AML. Any fatty tumor or focal fatty lesion of the liver that superficially resemble true lipomas should be tested for the presence of HMB-45 positive SMC in the tumor to differentiate it from AML. PMID- 10846575 TI - Nodular breast filariasis: diagnosis by fine needle aspiration. AB - Bancroftian filariasis is a tropical disease transmitted by mosquitoes that most often causes lymphadenitis. This article reports on a breast nodule due to Wuchereria bancrofti in a woman living in New Caledonia. The diagnosis was done by fine needle aspiration, which showed several microfilariae and avoided surgical treatment. PMID- 10846576 TI - Endometrial stromal sarcoma of the uterus with rhabdoid features. AB - A case of endometrial stromal sarcoma of the uterus with rhabdoid features, occurring in a 57 year old woman is reported. Electron microscopy and immunohistochemistry revealed that the rhabdoid cells contained intermediate filaments which were positive for vimentin, cytokeratin, alpha-smooth muscle actin, and muscle specific actin, but not for myoglobin and desmin. This indicated that the tumor in this case differed somewhat from the three rhabdoid tumors and an endometrial stromal sarcoma with rhabdoid differentiation previously reported and that, therefore, these tumors were heterogeneous. PMID- 10846577 TI - Phosphaturic mesenchymal tumor, mixed connective tissue variant (oncogenic osteomalacia). AB - A case of tumor-induced phosphaturic osteomalacia in a 54 year old man is reported. The patient was admitted because of progressive muscle spasms with pain and weakness in the bilateral thighs. Laboratory data showed hypophosphatemia, decreased tubular resorption of phosphate (TRP), a low 1,25-dihydroxyvitamin D level, and a high serum alkaline phosphatase level. Radiologic examinations revealed multiple lesions of osteomalacia in the ribs, and a small mass in the lower posterior mediastinum. After removal of the tumor, clinical symptoms disappeared and hypophosphatemia, decreased TRP, and the 1,25-dihydroxyvitamin D level were corrected. Microscopical examination revealed that the tumor was composed of mature adipose tissues, osseous tissues, and primitive stromal zones including osteoclast-like giant cells, non-mineralized woven bone, and various sized blood vessels. Patho-physiologic observations suggested that the tumor secreted some humoral substances inhibiting 25-hydroxyvitamin D-1 alpha hydroxylase activity, renal phosphate resorption, and parathyroid hormone production. PMID- 10846578 TI - [Continuing medical education and the professional career]. AB - The great scientific and technical advances in medicine make it necessary that the professional competence of the doctor needs to be brought up to date every now and then by the use of a continuing medical education (CME). The complexity and diversity of the professional subjects and the different areas of specialization give us reasons why the education of the doctor to-day should not be limited to the years of studying for a degree at university but that this education should be prolonged to include successive stages of specialized education and of the CME. The way of doing the latter (CME) is varied by means of different types of activities and it is recommended that it should be of a voluntary nature, through the personal motivation of the doctor, although he should also be spurred on because of different types of "prizes", as, for instance, social recognition, prestige, promotion to higher posts, etc. The organization of the activities of the CME should look to the recognition of the quality of the work by means of the corresponding accreditations according to uniform criteria established by a national central commission which should also establish the credits necessary for obtaining periodically the certificate for the bringing up to date of professional competence. This certificate, which should be gained in periods of 3 or 4 years duration, constitutes the basis, along with other elements (works of investigation, publications, years of tests, etc.) for the professional career of the doctor, the possible structure and development of which is shown in this presentation. PMID- 10846579 TI - [My interpretation as a pediatrician of "Las Meninas"]. AB - Las Meninas from Velazquez is one of the most important pictures in the world nevertheless, there is not an interpretation out of doubt of his meaning, in spite of all the studies written about it. After an original study of the history of Las Meninas, the persons painted, what are they doing, the Spanish culture on the time in what was painted and the historical life of the Infanta Margarita, the central personage of the picture, the author arrives to the conclusion about the possibility that she had a precocious puberty. PMID- 10846580 TI - [Why has menopause become a public health problem?]. PMID- 10846581 TI - [Hormone replacement treatment and breast cancer]. PMID- 10846582 TI - [The heme oxygenase system and its physiopathology]. AB - The HENE-OX system (isoforms 1, 2 and 3) is the key enzyme in the oxygen dependent HEME catabolism pathway and cleavage to HEME into biliverdin IX alfa, iron and carbon-monoxide (CO). The biliverdin reductase catalyzes biliverdin into bilirubin unconjugated what is converted in bilirubin conjugated, mono and biglucuronized for the activity of hepatic Uridindifosfo-glucuronil-transferase (UDFGT). The hyperbilirubinemias unconjugated can produce kernicterus. This should be treated with metalloporphyrins (Zn-Proto IX, Zn-Mesoporphyrin, Sn-Proto IX, Sn-Mesoporphyrin). Relations between HEM-OX/CO and NOS/NO systems. The induction of the HEME-OX system is a protective cellular mechanism against oxidative damage. Studies about the relations of the HEME-OX system with functions of various tissues (liver, kidney, cardiovascular and respiratory and hematological system, etc.). PMID- 10846583 TI - [The psychosocial roots of racism and xenophobia]. AB - The concept of race is only a human particularity. In spite of this, the racism, as a construction of imagination or a paranocial delusion or an ethnic attitude, is spread among the narcissist personalities and the environment dominated by nationalist or religious fanaticism and anyway generates collective violence. There is also a psychosocial racism almost always actual in the common human relations, appeared in form of positive attitudes towards similar others and negative attitudes towards different others. The best defence against these both phenomena are the attitudes of tolerance, understanding and liking. PMID- 10846584 TI - [Corino Andrade disease]. AB - Familial amyloidotic polyneuropathy (FAP) or Corino Andrade disease, is a very frequent amyloidosis but little known. We summarized the epidemiologic and clinical aspects of the Majorcan patients diagnosed since 1976, which constitute the fifth worldwide endemic focus of this degenerative disease, and we reported the scientific advances on FAP which are the basis to control this redoubtable degenerative process: easy diagnosis of patients and asymptomatic carriers, etiopathogenetic treatment and genetic counseling to reach the eradication. FAP must be better known because of the progressive incidence in Spain and varied symptomatology, to avoid confusions with other neurologic or extra-neurologic diseases. PMID- 10846585 TI - Increased incidence of indinavir nephrolithiasis in patients with hepatitis B or C virus infection. AB - MATERIALS AND METHODS: A HIV-1 patient database was scanned in March 1998, and 750 patients were identified who had received HAART including indinavir. Of these, 28 cases had nephrolithiasis; and 85 asymptomatic indinavir-treated patients were randomly selected as controls. The characteristics of cases and controls were compared by analysis of variance for quantitative parameters and by Fisher's exact test for classes. RESULTS: We observed a significant increase in the incidence of nephrolithiasis in patients co-infected with HIV-1 and either hepatitis C virus (HCV) (HCV RNA-positive) or hepatitis B virus (HBV) (HBs antigen-positive) (odds ratio and 95% confidence intervals: 2.8 and 1.1-7.7), whereas no significant differences were demonstrated between cases and controls with regard to age (42.4 +/- 8.0 versus 39.8 +/- 9.8 years), sex (male patients 70.4 versus 74.1%), duration of HIV-1 infection (8.6 +/- 3.1 versus 7.7 +/- 4.0 years), duration of indinavir treatment (16.1 +/- 5.8 versus 14.1 +/- 5.4 months), AST increase > or = 1.25 of normal (29.6 versus 25.9%), or ALT increase > or = 1.25 of normal (33.3 versus 22.4%). In co-infected patients, ALT increase (> or = 1.25 of normal), but not AST increase, at the time of indinavir initiation was statistically related to the occurrence of nephrolithiasis. CONCLUSIONS: We found a significant increase of nephrolithiasis incidence in patients co-infected with HIV-1 and HCV or HBV, which suggests that underlying multifactorial hepatic damage may limit liver catabolism of indinavir, and consequently increase its renal excretion and the risk of nephrolithiasis. Caution is therefore advised when initiating indinavir treatment in HIV patients with evidence of HBV or HCV infection. PMID- 10846586 TI - Prevalence of transmitted nucleoside analogue-resistant HIV-1 strains and pre existing mutations in pol reverse transcriptase and protease region: outcome after treatment in recently infected individuals. AB - We retrospectively studied 38 Italian recently HIV-1-infected subjects who seroconverted from 1994 to 1997 to investigate: (i) the prevalence of nucleoside reverse transcriptase inhibitors (NRTI)-related mutations at primary infection; (ii) the proportion of naturally occurring mutations in reverse transcriptase (RT) and protease regions of patients naive for non-nucleoside RT inhibitors (NNRTIs) and protease inhibitors (PIs); (iii) the drug-susceptibility to NRTIs and PIs in subjects with NRTI- and/or PI-related mutations; and (iv) the outcome of seroconverters treated with various NRTIs or NRTI/PI regimens. Baseline HIV-1 plasma viraemia and absolute CD4 count at baseline could not be used to distinguish patients with NRTI- and/or PI-related pre-existing mutations from those with wild-type virus (P = 0.693 and P = 0.542, respectively). The frequency of zidovudine-related mutations was 21% in the study period. The response to treatment was not significantly different in subjects with or without genotypic zidovudine-related mutations at primary infection (P = 0.744 for HIV-1 RNA and P = 0.102 for CD4 cells). Some natural variation (2.6%) was present within regions 98-108 and 179-190 of RT involved in NNRTI resistance. The high natural polymorphism in the protease region present in our patients was similar to that reported by others. In our study some PI-associated substitutions, thought to be compensatory in protease enzymatic function, could confer intermediate to high PI resistance. As discrepancies between genotypic and phenotypic results may exist in recent seroconverters, our data suggest that the role of transmitted NRTI- and PI-resistant variants remain to be fully elucidated in vivo. PMID- 10846587 TI - Persisting HIV-1 replication triggered by acute hepatitis A virus infection. AB - We report the case of two patients in whom acute hepatitis A was associated with a marked and prolonged increase in human immunodeficiency virus type 1 (HIV-1) viral load. Although in one patient the rise in HIV-1 RNA might also have been related to the interruption of antiretroviral therapy, we also observed a similar pattern in the other patient who had a stable undetectable plasma viraemia prior to acute hepatitis and never received treatment with anti-retrovirals. Our observation supports the hypothesis that immune activation that is induced by acute hepatitis A virus (HAV) infection may trigger HIV-1 replication. This highlights the importance of maintaining antiretroviral therapy throughout the acute phase of hepatitis A and of preventing HAV infection through active immunization. PMID- 10846588 TI - Possible fatal interaction between protease inhibitors and methamphetamine. PMID- 10846589 TI - Antiretroviral resistance testing comes of age. PMID- 10846590 TI - Principles of HIV resistance testing and overview of assay performance characteristics. AB - HIV drug resistance testing is becoming an integral part of antiretroviral drug development and of patient management. The parameters that characterize the performance of both phenotypic and genotypic assays and the validation of this performance are essential to their proper use for these applications. Several principles and biological factors impacting drug resistance testing are summarized. Considerations regarding quality control and validation of the performance of genotypic and phenotypic assays are also addressed. PMID- 10846591 TI - Transmission and prevalence of HIV resistance among treatment-naive subjects. AB - Recent reports demonstrate the transmission of drug-resistant HIV-1 variants to newly infected individuals, although estimates of the prevalence of drug resistance among populations from different geographic regions are highly varied. The interpretation and comparison of available study results are confounded by the lack of consensus regarding the nomenclature and reporting of antiviral resistance. This report re-evaluates previously presented and published data using uniform criteria for genotypic and phenotypic drug resistance. Treatment inexperienced and recently or chronically infected populations are reviewed. The prevalence of transmitted drug resistance ranges from 1% to 11% among recently infected persons using these criteria. Programmes to monitor and characterize drug resistance among newly infected persons and their source partners are essential to evaluate the selection processes that influence the transmission of certain genetic variants of HIV. Temporal trends in the prevalence of transmitted drug resistance among diverse populations are necessary to evaluate the potential need for selective and generalized drug resistance screening programmes among newly infected, treatment-naive patients. PMID- 10846592 TI - The relation between baseline HIV drug resistance and response to antiretroviral therapy: re-analysis of retrospective and prospective studies using a standardized data analysis plan. AB - To assess the relation between resistance to antiretroviral drugs for treatment of HIV-1 infection and virological response to therapy, results from 12 different studies were re-analysed according to a standard data analysis plan. These studies included nine clinical trials and three observational cohorts. The primary end-point in our analyses was virological failure by week 24. Baseline factors that were investigated as predictors of virological failure were plasma HIV-1 RNA, the number and type of new antiretroviral drugs in the regimen, and viral susceptibility to the drugs in the regimen, determined by genotyping or phenotyping methods. These analyses confirmed the importance of both genotypic and phenotypic drug resistance as predictors of virological failure, whether these factors were analysed separately or adjusted for other baseline confounding factors. In most of the re-analysed studies, the odds of virological failure were reduced by about twofold for each additional drug in the regimen to which the patient's virus was sensitive by genotyping methods, and by about two- to threefold for each additional drug that was sensitive by phenotyping. PMID- 10846593 TI - HIV drug susceptibility and treatment response to mega-HAART regimen in patients from the Frankfurt HIV cohort. AB - OBJECTIVE: To assess the relationship between viral susceptibility at baseline and virological response in human immunodeficiency virus (HIV)-infected patients treated with multi-drug salvage regimens after multiple previous treatment failures. DESIGN: Retrospective analysis of 50 patients from the Frankfurt HIV cohort who had received treatment with a minimum of six drugs, and for whom a sample for baseline viral phenotyping was available. METHODS: Viral drug susceptibility was measured retrospectively from stored samples using the Antivirogram, a recombinant virus assay based method. Virological response was defined as a viral load of < 400 copies/ml at week 24. For analysis of treatment response, drop-outs were dealt with in two ways, either as failures (DAF) or censored (DAC). Several logistical regression models were applied to identify predictors of response, including baseline virus load, number of new drugs and phenotypic sensitivity scores. RESULTS: At baseline, drug resistance was extensive: 96% of patients had viruses resistant to at least one drug class and 32% had viruses resistant to all three drug classes. In the DAF analysis, 39 patients experienced virological failure. In the DAC analysis, eight were censored and 31 patients experienced virological failure. In multivariate models that adjust for baseline viral load, the number of new drugs and total phenotypic sensitivity scores, the baseline viral load and phenotypic sensitivity score remained significantly associated with virological outcome, whereas in those adjusted for baseline viral load, the number of new drugs, NRTI phenotypic sensitivity score and PI phenotypic sensitivity score, only the latter remained significantly associated with virological outcome. Both the DAF and DAC analyses produced similar results. In all models used, virological failure was shown to be significantly associated with baseline viral load and phenotypic sensitivity score. CONCLUSIONS: In this retrospective analysis based on a small number of patients, viral drug susceptibility at baseline was strongly associated with virological outcome at 24 weeks, independent of covariates such as baseline viral load and treatment history. Baseline viral load also maintained a significant, independent association with virological outcome in most models. PMID- 10846594 TI - Frequency of antiretroviral drug resistance mutations in HIV-1 strains from patients failing triple drug regimens. The Terry Beirn Community Programs for Clinical Research on AIDS. AB - The frequency of protease and reverse transcriptase (RT) gene mutations was determined in HIV-1 strains from 153 patients entering the CPCRA 046 (GART) study who were failing triple-drug regimens consisting of one protease inhibitor (PI) and two RT inhibitors. Population-based sequence analyses showed that nearly all patients had similar RT gene mutations regardless of prior drug exposure, although the M184V mutation was significantly less prevalent in patients not recently treated with lamivudine. Whilst typical inhibitor-specific ('signature') protease gene mutations were found in patients failing their first PI, these mutations were significantly less likely to be found in patients exposed to two or more PIs. Protease gene mutations associated with multi-PI resistance were more likely to be observed in patients treated with more than one PI. These results suggest sequential treatment with PIs select for a relatively limited number of protease gene mutations that likely originated during early PI therapy. These protease gene mutations and a similarly limited set of RT gene mutations appear to be responsible for treatment failure in antiretroviral therapy. PMID- 10846595 TI - Persisting long-term benefit of genotype-guided treatment for HIV-infected patients failing HAART. The Viradapt Study: week 48 follow-up. AB - OBJECTIVE: We report the 12 months follow-up of the patients who participated in the Viradapt study. METHODS: A total of 108 HIV-infected patients failing antiretroviral (ARV) therapy (HIV RNA > 10,000 copies/ml, therapy > 6 months with nucleoside reverse transcriptase inhibitors, > 3 months with protease inhibitors (PIs) were randomized into two arms: standard of care in the control arm, and treatment according to the resistance mutations in the protease and reverse transcriptase genes in the study arm. After the first 6 months of the randomized study, open-label, genotype-guided treatment was offered in both arms. A multivariate analysis was performed to assess the predictive factors of treatment success (HIV RNA < 200 copies/ml). RESULTS: The two arms were comparable in terms of risk factors, age, sex, previous treatments, CD4 cell count and log10 HIV-1 RNA at baseline. At week 24, an interim combined analysis showed a statistically significant difference in the drop in viral load at months 3 and 6 (P = 0.015, repeated measures analysis of variance) in favour of the genotype group. Patients in both arms were then offered open-label genotyping. Genotype analysis was performed every 3 months, and treatment changes could accordingly be made. As some of the patients in the control arm had already progressed to months 9 or 12, only 69% (30/43) of these patients received genotype-guided treatment changes. In the genotype arm, the mean drop in HIV RNA of 1.15 log10 copies/ml, obtained at month 6, persisted at months 9 and 12 (1.15 log10 copies/ml +/- 0.17). In the control arm, an additional drop in HIV RNA to 0.98 log10 +/- 0.22 copies/ml was observed by month 12. In control patients receiving open-label genotype, the percentage of patients with HIV-1 RNA levels below detection limit (200 copies/ml) rose from 14% at month 6 to 30.5% at month 12. This percentage in the study arm remained stable at 31.3% and 30% at months 9 and 12, respectively. Genotype-guided therapy, primary protease mutations and PI plasma concentrations were significantly correlated with virological success. CONCLUSION: In this heavily pretreated patient population, genotype-guided therapy resulted in a sustained reduction in HIV RNA of greater than one log10 throughout a 1 year follow-up period. Performance of genotype-guided therapy may have contributed to the additional viral load reduction seen in patients in the control group who received open-label genotyping after the 6 months point. Multivariate analysis showed that the presence of primary protease mutations, performance of genotype guided treatment changes and PI plasma concentrations independently affected virological response. PMID- 10846596 TI - Limits of resistance testing. AB - Interpretation of antiretroviral testing is complicated by many factors, including those related to HIV biology and genetics, as well as the intricacies of drug selection pressure in vivo. These complex factors can limit the usefulness of resistance testing. However, knowledge about these issues can help to avoid misinterpreting resistance test results and thereby help clinicians to use resistance testing to individualize antiretroviral drug choices. PMID- 10846597 TI - The role of resistance testing in clinical trial design and product labelling: a regulatory perspective. AB - Assays that attempt to characterize HIV susceptibility or resistance are among the latest technologies that are likely to impact HIV clinical trial design, antiretroviral drug development and patient management. However, at present the Food and Drug Administration (FDA) have yet to approve any phenotypic or genotypic HIV resistance assay and the role of resistance testing in clinical management of patients and in drug development is ill defined. In November 1999, the Division of Antiviral Drug Products at the FDA convened a meeting of its advisory committee to consider the available information about HIV resistance testing, and to generate some recommendations about how these assays could be utilized in antiretroviral drug development. In addition, the committee was presented with several hypothetical regulatory scenarios in order to illustrate how HIV resistance testing might be incorporated in antiretroviral drug development and drug labelling. In this article, we discuss the regulatory history of resistance testing in antimicrobial drug development, the current use of resistance testing for antiretrovirals, as well as a summary of the hypothetical scenarios that were presented to the committee and the discussion of the committee members regarding those scenarios. PMID- 10846598 TI - [Inhalation or oral therapy in the treatment of asthma]. PMID- 10846599 TI - [Features of patients with bronchial asthma seen at the emergency department of a referral hospital in a semirural area]. AB - Bronchial asthma is a chronic disease whose prevalence and severity is increasing. Appropriate treatment of exacerbation seems to affect the subsequent course of disease. OBJECTIVES: To assess the extent of application of guidelines for treating exacerbations of asthma and to describe the clinical and epidemiological characteristics of patients. PATIENTS AND METHODS: All patients presenting with exacerbations to the emergency room of a referral hospital in a semi-rural area of Catalonia were enrolled consecutively. We recorded symptoms, function and treatment variables during the emergency and when the patient was stable, at which time we also administered the Asthma Quality of Life Questionnaire (AQLQ). RESULTS: Forty-seven percent of patients resided in rural villages. Seventy-one percent lived in the area served by our hospital. Of the remaining patients, 57% came from municipalities in the Baix Emporda area, 29% in Alt Emporda area and 14% in La Selva area. Seventy-one percent went to the hospital directly from home, 66% of them were in the province of Girona; 75% of those homes were within urban areas. Of the 29% who had been sent to the emergency room by a physician, 71% were from a rural area. The 119 asthma exacerbations treated (114 patients, 71 females, 42 +/- 23 years) accounted for 0.3% of all emergency room visits. The exacerbation was considered severe in 31%, moderate in 47% and mild in all remaining cases. Eighty-eight percent of patients had experienced an exacerbation before. Symptoms were often present before the attack, but nearly half the patients were receiving relatively inadequate medication. Peak expiratory flow and oxygen saturation were recorded in 70% of cases and both variables improved after initial treatment (250 +/- 97 to 349 +/- 92 l/min, p < 0.001; and 92 +/- 7 to 96 +/- 2%, p < 0.01, respectively). All patients received inhaled salbutamol, 44% inhaled corticosteroids and 73% intravenous corticosteroids. A quarter of the patients were admitted to the ward (0.4% of all admissions) and 4% were admitted to the intensive care unit. Patients stayed 8.8 +/- 1.4 h in the emergency room. When patients were stable, asthma was severe in 14%, moderate in 42%, mild but persistent in 27%, and occasional in 17%. The total score on the AQLQ was 13.8 +/- 11.1, with mood being the dimension with the highest score (5.1 +/- 4.9). This score, along with social restriction, were lowest in the group of patients with chronic airflow obstruction. CONCLUSIONS: Nearly half the patients were from rural villages most of which were located outside the zone served directly by our hospital. The preference for specialized medical attention would explain the percentage of patients seen who did not belong to the assigned area. Difficulty of access to the hospital from certain rural areas would account for the greater number of rural patients who had been referred by a physician. Clinical management and monitoring of asthmatic patients with exacerbation could be improved by greater diffusion and application of guidelines. It would be interesting to incorporate come sort of questionnaire on quality of life while taking a patient's history. PMID- 10846600 TI - [Validation of half-night polysomnography for sleep apnea/hypoapnea syndrome]. AB - A diagnosis of sleep apnea/hypopnea syndrome (SAHS) is based on clinical signs and nighttime polysomnograms. Brief polysomnography has been proposed as an alternative to all-night recording. OBJECTIVES: The aim of this study was to determine whether a polysomnograms obtained during the first half of the night is sufficient for establishing a diagnosis of SAHS and to determine the correlation between polysomnographic variables recorded during the first four hours (half the study time) with those recorded over the full eight hours (full study time), as well as to determine diagnostic agreement. DESIGN: Thirty-five patients suspected of having SAHS were studied prospectively. Baseline polysomnograms were scored blindly by two independent observers following standard methods. A diagnosis of SAHS was made according to guidelines of the Spanish Society of Pneumology and Chest Surgery. During the first half of the night and up to the end of each recording period we gathered neurophysiological and respiratory variables and diagnostic impressions. RESULTS: The correlation between variables (sleep stage, overall AHI, REM-AHI, non-REM-AHI and sleep efficiency) recorded in the first half of the night and throughout the night was significant (p < 0.05) by both Pearson's correlation coefficient (r) and by the intraclass correlation coefficient (ICC). In 33 of 35 patients (94.3%) diagnostic agreement was achieved (95% CI 80.84-99.30); when SAHS was severe, agreement was 100%. CONCLUSION: Based on these results, we conclude that for patients with a diagnosis of severe SAHS during the first half of the night, data recorded during the second half can be considered supplementary. PMID- 10846601 TI - [Respiratory disease in young people and smoking]. AB - OBJECTIVE: To assess the prevalence of respiratory disease among adolescents and young adults and to determine whether disease is more common among smokers. POPULATION AND METHOD: Four hundred twenty subjects (45.3% female and 54.7% male) between 14 and 21 years of age were studied. The subjects were enrolled in public and private schools in Tenerife (Spain). Each subject filled in an epidemiological questionnaire as recommended by the American Thoracic Society (ATS). RESULTS: We found that 26.5% were habitual smokers, 11.0% were occasional smokers, 3.6% were ex-smokers and 45.6% were passive smokers. Only 13.4% reported not being exposed to tobacco smoke. Respiratory diseases included recurrent upper respiratory tract infections (14.1%), lower respiratory tract infections (16%), allergy (32.5%) and asthma (10.5%), bronchospasm with exercise (25.8%) and chronic coughing (8.6%). The incidences of the following diseases were higher in young smokers: upper respiratory tract infections (p = 0.001), lower respiratory tract infections (p = 0.004), bronchospasm with exercise (p = 0.02) and chronic cough (p = 0.0001). Young smokers also lost more school days per year due to respiratory disease (p = 0.01). The rate of lower respiratory tract infections per year was higher among young smokers than among those who were unexposed (p = 0.004). CONCLUSION: Smoking, which leads to a higher incidence of respiratory disease, has a negative impact on adolescents. PMID- 10846602 TI - [Tuberculosis management unit: 7-year experience]. AB - Between January 1992 and December 1998 we collected clinical, epidemiological and treatment data on all patients diagnosed of tuberculosis in our specialized unit. Five hundred sixty-seven patients (70% male and 30% female) were studied prospectively. The rate of new cases increased until 1995 and decreased during the last three years of study. Mean patient age was 38.8 years, with nearly 64% of patients under 45 years of age. Predisposing disease, mainly chronic alcoholism, was present in 36%. Fifteen percent belonged to a high-risk social group (6.5% were drug addicts and 6.3% lived inside secure institutions). The mean time elapsing from the appearance of symptoms until referral to our service was 80.4 days and the most common clinical picture at presentation was general unwellness with cough and expectoration (46%) followed by hemoptysis (18%). Cavitation was visible in 48.5% of x-rays, while alveolar infiltrates were seen in 33%, pleural effusion in 12% and lymph node involvement in 10%. Adult tuberculosis was diagnosed in 80% of cases, 10% were reactivations and 9% were primary. Bacteriological diagnosis was available for 85%. Therapy usually involved six months with hydrazide, rifampicin and pyrazinamide (81%). Therapy was generally well-tolerated, although analyses revealed some anomalies, such as transaminase alteration (18%) and hyperuricemia (19%). Therapy was changed because of toxicity in only 2.6%. Follow-up after therapy was strict and the rate of successful cure was 97.5%. We conclude that diagnosis was not prompt enough and believe that knowledge of epidemiological, clinical and evolutionary data, as well as monitoring of real rates of cure of treated patients justifies the existence of specialized centers for managing tuberculosis. PMID- 10846603 TI - [Estimating forced expiratory volume in one second based on breath holding in healthy subjects]. AB - Cross sectional study to evaluate a linear correlation model to calculate forced expiratory volume in one second (FEV1) on the basis of breath-holding time (BHT) in healthy subjects aged 16 years or older. BHT was defined as the maximum time in seconds that a person can voluntarily hold his or her breath measured between the end of a deep breath until he or she begins to exhale in the third of three consecutive maneuvers. To calculate FEV1 on the basis of BHT (FEV1-BHT) the general straight-line equation (y = a + bx) was applied, with x being Logn of BHT and y being FEV1. The a and b values had been previously estimated by gender using multiple linear regression in which FEV1 measured spirometrically was the dependent variable and the age and Logn of BHT were the independent variables. One hundred ninety-seven subjects [97 women (49.2%) and 100 men (50.8%)] were enrolled. Mean FEV1 measured by spirometry (FEV1-Sp) and FEV1-BHT were similar. The correlation (R2) between FEV1 and BHT for women and men was 0.585 (95% CI 0.213-0.753) and 0.702 (95% CI 0.630-1.496), respectively. The ratio FEV1 BHT:FEV1-Sp was higher than 80% for 96 women (98.96%) and 98 men (98.0%). FEV1 can be reliably estimated using BHT. PMID- 10846604 TI - [Respiratory muscle structure in healthy subjects and changes in patients with chronic obstructive pulmonary disease]. PMID- 10846605 TI - [Study and diagnosis of chronic cough in adults]. PMID- 10846606 TI - [Mediastinal hydatid cysts: 8 cases]. AB - Hydatidosis in humans is a parasitic disease produced by cystic growth of the larvae of the tapeworm Echinococcus granulosus, affecting mainly the liver and lung. Larvae are rarely present in the mediastinum, although approximately 100 cases have been described in the literature. We report eight cases of hydatid cysts of the mediastinum (HCM) treated surgically over a period of 21 years. The incidence was similar in males and females and ages ranged from 10 to 74 years. Symptoms depend on size, location and involvement of neighboring structures in HCM. The most serious complication is cyst rupture with consequent transfer of hydatid material to the blood, possibly causing anaphylactic shock and even death. Currently, sonography, computed tomography and magnetic resonance images facilitate diagnosis. Treatment involves excision of the cyst and peri-cystic tissue. CHM should be suspected when mediastinal cysts are found in countries where the incidence of hydatidosis is high. PMID- 10846608 TI - [Quinolone toxicity]. PMID- 10846607 TI - [Primary ciliary dyskinesia. A new phenotypic variant]. AB - Primary ciliary dyskinesia (PCD) is a genetic disorder characterized by the inability of ciliated structures to beat effectively. Clinical course includes recurrent sinus and ear infections, chronic or recurrent bronchitis and infertility in men. Although several phenotypes have been described, lung function deterioration secondary to bronchiectasis becomes severe only rarely. That upper airway tract infections go unnoticed has not been reported. We report a case of PCD characterized by immotile sperm, severe obstructive respiratory disorder that required a sequential double lung transplant with the absence of recurrent sinus and ear infections. PMID- 10846609 TI - [Pulmonary lymphangioleiomyomatosis associated with epilepsy. A step further toward tuberous sclerosis?]. PMID- 10846610 TI - [Bilateral tuberculous pleural effusion and HIV infection]. PMID- 10846611 TI - [Toxic neutropenia caused by antitubercular drugs]. PMID- 10846612 TI - [Spontaneous pneumomediastinum in a cocaine user]. PMID- 10846613 TI - Effects of a nitric oxide synthase inhibitor on pressor response to angiotensin II in conscious rats. AB - OBJECTIVE: We sought to characterize the effects of NG-monomethyl-L-arginine (L NMMA) and L-arginine (L-Arg) on the pressor response to the infusion of angiotensin II in rats. METHODS: L-NMMA and L-Arg were infused intraperitoneally into rats at a constant rate by means of an osmotic minipump. The L-NMMA group received an infusion of L-NMMA (3 mg/d) daily for 13 d, whereas the L-NMMA plus L Arg group received L-NMMA (3 mg/d) daily for 4 d, followed by L-NMMA plus L-Arg (12 mg/d) daily for 9 d. Sham operated rats served as controls. The animals were anesthetized on day 13, and catheters were placed into the femoral artery and vein. After the animals had recovered from the anesthesia, the pressor response to intravenous bolus doses of angiotensin II (50, 100, 200, and 400 ng/kg) were determined after recovery from anesthesia. RESULTS: While the baseline mean arterial blood pressure was not affected by L-NMMA, with or without L-Arg, the pressor response to angiotensin II in the L-NMMA group was significantly increased as compared with that in the control group, at doses of 50, 100, and 200 ng/kg. The response of the L-NMMA plus L-Arg group did not differ significantly from that of the control group. CONCLUSION: Results indicate that the infusion of a nitric oxide synthase inhibitor, at a dose insufficient to produce hypertension, increases the pressor response to angiotensin II. PMID- 10846614 TI - Aldehyde induced hypertension in rats: prevention by N-acetyl cysteine. AB - Methylglyoxal, a highly reactive endogenous aldehyde is formed in the tissue of humans and animals as an intermediate of glucose and fructose metabolism. N acetyl cysteine (NAC), an analogue of the dietary amino acid cysteine, binds aldehydes thus preventing their damaging effect on physiological proteins. We measured systolic blood pressure (SBP), platelet [Ca2+]i, circulating nitric oxide levels, tissue aldehyde conjugates and renal vascular changes in chronic methyglyoxal treated Wistar-Kyoto (WKY) rats and examined the effect of NAC in the diet on these parameters. Animals, age seven weeks, were divided into three groups of six animals each and were treated as follows: WKY-control (chow diet and normal drinking water); WKY-methylglyoxal (chow diet and methyglyoxal in drinking water); WKY-methyglyoxal + NAC (1.5% NAC in diet and methylglyoxal in drinking water) for the next 18 weeks. Methylgyoxal in drinking water was given at a concentration of 0.2% during weeks 0-5; 0.4%, weeks 6-10; and 0.8%, weeks 11 18. After 18 weeks systolic blood pressure, platelet [Ca2+]i and kidney aldehyde conjugates were significantly higher and serum nitric oxide levels lower in methylglyoxal treated rats. Methylglyoxal treated rats also showed smooth muscle cell hyperplasia in the small artery and arterioles of the kidney. N-acetyl cysteine, an aldehyde binding thiol compound, prevented these changes. PMID- 10846615 TI - Effect of aspirin on the contractility of aortic smooth muscle and the course of blood pressure development in male spontaneously hypertensive rats. AB - The effects of acetylsalicylic acid (ASA) on aortic smooth muscle contractility were studied in aortic rings of male SHR and WKY rats. The rats were administered two intraperitoneal injections of 10 mg/kg of ASA per week for ten weeks. Blood pressure of each rat was monitored twice weekly prior to the i.p. injections. Twenty four hours after the last injection the aortic smooth muscles were evaluated for generation of active tension in response to KCl, Phenylephrine (PE), Clonidine and Norepinephrine (NE). In another set of experiments calcium conductance was evaluated in the presence or absence of endothelium both in ASA treated and non treated animals. We report that aortic rings from ASA-treated SHR animals were more responsive to contractile agents than rings from non-treated SHR male rats. Also, the Ca2+ conductance in vitro was enhanced appreciably in SHR aortic rings denuded of their monolayer of endothelium in response to ASA treatment. No decrease in systolic blood pressure was observed in response to ASA treatment in SHR male rats. These results suggest that acetylsalicylic acid not only may modulate aortic smooth muscle contractility through the metabolites of arachidonic acid but may repair to a great extent the hypertension associated plasma membrane permeability defect of vascular myocytes. PMID- 10846616 TI - Comparative genomics of chalcone synthase and Myb genes in the grass family. AB - Most plant genes occur as members of multigene families where new copies arise through duplication. Duplicate genes that do not confer an adaptive advantage to the plant are expected to rapidly erode into pseudogenes owing to the accumulation of transpositions, insertion/deletion mutations and nucleotide changes. Nonfunctional copies will drift to fixation within a few million years and ultimately erode beyond recognition. Duplicate genes that are retained over longer periods of evolutionary time must be positively selected based on some adaptive advantage conferred on the plant species. We explore the dynamics of the recruitment of new duplicate genes for chalcone synthase, the enzyme that catalyzes the first committed step of flavonoid biosynthesis, and for the myb family of transcriptional activators. Our analyses show that new chs genes are recruited into the genome of grasses at a rate of one new copy every 15 to 25 million years. In contrast, the myb gene family is much older and many duplicate copies appear to predate the separation of the angiosperm lineage from other seed plants. The general pattern suggests a rapid adaptive proliferation of new chs genes but a more ancient elaboration of regulatory gene functions. Our analyses also reveal accelerated rates of protein evolution following gene duplication and evidence is presented for interlocus exchange among duplicate gene loci. PMID- 10846617 TI - Molecular phylogeny of East Asian moles inferred from the sequence variation of the mitochondrial cytochrome b gene. AB - Taxonomic analysis has previously revealed that the species of moles that inhabit Japan are characterized by exceptional species richness and a high level of endemism. Here, we focused on the evolutionary history of the four Japanese mole species of the genera Euroscapter and Mogera, examining mitochondrial cytochrome b (cyt b) gene sequences and comparing them with those of continental Mogera wogura (Korean and Russian populations), M. insularis from Taiwan, and Talpa europaea and T. altaica from the western and central Eurasian continent, respectively. Our data support the idea that in a radiation center somewhere on the Eurasian continent, a parental stock evolved to modern mole-like morph and radiated several times intermittently during the course of the evolution, spreading its branches to other peripheral geographic domains at each stage of the radiation. Under this hypothesis, the four lineages of Japanese mole species, E. mizura, M. tokudae, M. imaizumii, and M. wogura, could be explained to have immigrated to Japan in this order. Mogera wogura and M. imaizumii showed substantial amounts of geographic variation and somewhat complicated distributions of the cyt b gene types. These intraspecific variations are likely to be associated with the expansion processes of moles in the Japanese Islands during the Pleistocene glacial ages. PMID- 10846618 TI - Determination of the complete nucleotide sequence and haplotypes in the D-loop region of the mitochondrial genome in the oriental white stork, Ciconia boyciana. AB - The complete nucleotide sequence of the mitochondrial genome of the Oriental white stork, Ciconia boyciana, has been determined from captive storks by a novel method incorporating Long PCR and shotgun sequencing. 13 protein-coding genes, two ribosomal RNA genes and 22 transfer RNA genes were identified as in other vertebrate mitochondrial genomes. The position and direction of the NADH6 and tRNA-Glu genes were the same as previously reported for avian mitochondrial genomes. A 71 bp direct repeat and long CAAA repeat sequences were found at the 3' end of the D-loop region, together with SCB-1, SCB-2, SCB-3, and three TAS sequences. Direct sequencing of the PCR fragments in the D-loop region in 26 captive Oriental white storks originating from Japan, China, and Russia revealed nucleotide differences at 18 sites along 1,248 bp, and a total of nine haplotypes have been identified. It was found that one pair of individuals in the Japanese captive breeding program were of the same haplotype, suggesting that they were caught from the same nest. The pair has since been dissolved in consideration of the possibility of inbreeding depression. PMID- 10846619 TI - An improved integration replacement/disruption method for mutagenesis of yeast essential genes. AB - We improved the integration replacement/disruption method (Shortle, D., Novic, P., and Botstein, D. Proc. Natl. Acad. Sci. USA 81: 4889-4893, 1984) for isolating mutants in any of essential genes of the yeast Saccharomyces cerevisiae by integrating mutagenized DNA into the wild type gene of interest. We adopted this method to isolate temperature-sensitive mutants of the MPC1 gene encoding the YLL031C ORF. To facilitate integration of the mutagenic plasmid at a site near the 5' end of the ORF, a BamHI site was created at 300 bp downstream of the 5' end of the truncated ORF to be mutagenized. The MPC1 gene was disrupted in the wild type haploid strain by integrating a 5'-truncated derivative of the gene with mutations induced by in vitro mutagenesis. Transformants thus obtained were subjected for diagnosis of conditional lethality by replica-plating onto an appropriate selection medium to detect mutants. A primary mutant isolated by this method reverted in a high frequency due to a tandem repeat created by mutagenic integration. We deviced a method to obtain a stable temperature-sensitive strain by disrupting the tandem duplication. Two stable temperature-sensitive mutants thus obtained were found to be remedial either with 1 M sorbitol or with 0.1 M Mg2+ and to be sensitive to local anestheticum, tetracaine, at 25 degrees C. PMID- 10846620 TI - A site specific increase in recombination in Drosophila ananassae. AB - The e65 pi; bri ru stock of Drosophila ananassae produced an extremely high rate of recombination in males when made heterozygous with any one of the wild type stocks. We analyzed and characterized the genetic factors which caused this phenomenon. We show that the second chromosome of the e65 pi; bri ru stock carries an enhancer of male recombination. The enhancer, En(2)-ep, is located between Om(2C) and Arc. The enhancement of meiotic recombination both in males and females was also observed at the specific region between Om(2C) and Arc on 2L. The magnitude of increased recombination was 30-40 fold in males and 13-30 fold in females. The relation between the hotspot of recombination in both sexes and the enhancer of male recombination is discussed. PMID- 10846621 TI - A cold-responsive wheat (Triticum aestivum L.) gene wcor14 identified in a winter hardy cultivar 'Mironovska 808'. AB - A cDNA library was constructed from a cold-acclimated winter-hardy common wheat (Triticum aestivum L.) cultivar 'Mironovska 808'. Using this library and a cold- and light-responsive barley cDNA clone cor14b as a probe, cDNAs of a homologous wheat gene wcor14 were isolated. Two identical cDNAs designated as wcor14a had an open reading frame encoding an acidic (pI = 4.71) and hydrophobic polypeptide with 140 amino acids (MW = 13.5 kDa). The deduced WCOR14a polypeptide showed 70% identity with the barley chloroplast-imported COR14b and had a nearly identical N terminal, putative chloroplast transit peptide of 51 amino acid residues. Another cDNA clone wcor14b was assumed to encode a polypeptide WCORb which had 5 substitutions and a frame shift in the C-terminal region as compared with WCOR14a. RACE PCR, genomic PCR and Southern blot analyses suggested that wcor14 and its related sequences constitute a small multigene family with and without an intron in the hexaploid wheat genome. Northern blot analysis showed that transcripts of wcor14 accumulated within 3-6 hours of cold acclimation at 4 degrees C and the level reached a maximum at day 3. The transcripts became non detectable within 3 hours after de-acclimation at room temperature. Contrary to the barley cor14b, a similar level of wcor14 transcripts was detected under the continuous darkness. Neither treatment with NaCl, ABA nor dehydration induced its expression. Based on these results we conclude that wcor14 is a wheat orthologue of the barley cor14b and specifically induced by low temperature. PMID- 10846622 TI - Beyond IQ: broad-based measurement of individual success potential or "emotional intelligence". AB - Individual-difference correlates of life success were studied using 302 participants aged 17-46 years (107 men, 195 women) and 31 individual-difference measures. Factor 1 (Relaxed Temperament) included Trait Pleasure-Displeasure (+), Anxiety (-), Depression (-), Optimism (+), Self-Esteem (+), Covert Index of Employee Reliability (+), and Functional Flexibility (-). Positively loading scales on Factor 2 (Arousable Temperament) were Trait Arousability, Emotional Empathy, Emotional Thinking, and Affiliative Tendency. Factor 3 (Disciplined Goal Orientation) included Delay of Gratification (+), Impulsivity (-), Procrastination (-), Patience (+), Integrity (+), Adaptive Coping (+), and Intelligence (+). Positively loading scales on Factor 4 (Dominant Temperament) were Trait Dominance-Submissiveness, Social Competence, Achieving Tendency, and Self-Actualization. Factors 1, 3, 4, and intelligence exhibited positive relations with all peer-rated criterion measures of life success. Trait Arousability, representing Factor 2, correlated negatively with relationship, physical, work, and overall success. Physical attractiveness correlated positively with all success measures except emotional success. When success measures were regressed against intelligence and personality scales or factors, intelligence did not account for variance beyond that explained by personality. PMID- 10846623 TI - Toward the differentiation of trust in supervisor and trust in organization. AB - Trust in supervisor and trust in organization are argued to be distinct but related constructs, each with its own set of antecedents and outcomes. Empirical field results supported the proposition. Although trust in supervisor and trust in organization were positively and significantly correlated, trust in supervisor was more strongly associated with proximal variables (ability, benevolence, and integrity of supervisor), whereas trust in organization was more strongly correlated with global variables (perceived organizational support and justice). This conclusion held despite the inclusion of proximal variables in the regression on trust in organization and the inclusion of global variables in the regression on trust in supervisor. In addition to the differential antecedents of trust in supervisor and trust in organization, the outcomes for both variables were different. Trust in supervisor was related to increased innovative behavior and satisfaction with supervisor, and trust in organization was related to higher organizational commitment and lower intention to leave. Therefore, the authors provide clear preliminary data on the distinctiveness of trust in supervisor and trust in organization. One implication of this set of results is that organizations should adopt a more holistic approach in building trust, which can be achieved by focusing on the various constituents of the organization and the various levels (e.g., the supervisor level and the organizational level). PMID- 10846624 TI - Impact of Human Genome Project on biological sciences. PMID- 10846625 TI - Comparative evaluation of different rat models with co-existing diabetes-mellitus and hypertension. AB - We have evaluated the suitability of different rat models for the study of effects of antihypertensives on cardiovascular and metabolic complications of diabetes mellitus and hypertension. IDDM was induced in Wistar and spontaneously hypertensive (SH) rats by single tail vein injection of STZ (45 mg/kg, i.v.). Neonatal STZ-diabetes (nSTZ) was induced by administering STZ, 70 mg/kg (i.p.) to 5 day old Wistar rat pups. DOCA-hypertension was induced in Wistar and STZ diabetic rats using deoxycorticosterone acetate (DOCA, 5 mg/kg, s.c.) and NaCl (2%) in drinking water. Intravenous injection of STZ produced cardinal signs of diabetes mellitus including hyperglycemia, loss of body weight, polyphagia and polydipsia. STZ-diabetic rats also showed hyperlipidemia and hypoinsulinemia. STZ treated rats developed hypertension and bradycardia. nSTZ rats were found to have mild hyperglycemia and were hypertensive and hyperinsulinemic. The OGTT and ITT revealed that nSTZ rats are insulin resistant. SH rats were also found to be hyperinsulinemic and hypertensive. Although, these rats were found to be insulin resistant, they did not demonstrate hyperglycemia. DOCA-treated STZ-diabetic rats were found to have milder hyperglycemia when compared to STZ-diabetic rats not treated with DOCA. Although, DOCA treatment was not found to alter serum levels of glucose and insulin, results of OGTT revealed enhanced glucose disposal in DOCA-treated Wistar rats, suggesting that DOCA probably produces some effect on glucose homeostasis in rats. The present data also suggest that STZ-diabetic rat may be considered a suitable model for IDDM. On the other hand, nSTZ and SH rats were hyperinsulinemic and insulin resistant and may be used as models to study insulin sensitivity. DOCA-hypertensive rat may not be a suitable model for studying the effects of various drug interventions on glucose homeostasis and insulin sensitivity as DOCA itself appears to influence these factors. PMID- 10846626 TI - Neuro-immunomodulation by dorsolateral hippocampus--role of macrophages,T and B cells. AB - In dorsolateral hippocampal lesioned, Sham and control animals, the total number of splenocytes was determined and macrophages, B cells and T cells were isolated and their percentage distribution in total splenocytes were determined. The leukocyte migration inhibition was studied in these groups using different fractions of splenocytes namely whole splenocytes, macrophage removed fraction, and nylon wool eluted T cell population. The same groups were also studied after an antigenic challenge. The results indicate alterations in the macrophages, adherent cell population as well as T cell population in lesioned and Sham animals and also in their immunized groups. These altered cell sub-populations may be responsible for the altered migration. PMID- 10846627 TI - Neuromuscular blocking effects of an alkaloidal extract from Inula royleana: contractile and electrical studies on amphibian skeletal muscle in vitro. AB - The neuromuscular blocking properties of an alkaloidal extract from the root of Inula royleana have been investigated in vitro using a combination of mechanical and electrophysiological approaches. Neurogenic twitches of the frog sartorius were profoundly inhibited by concentrations of the extract > or = 20 micrograms/ml, being reduced to 50% of control amplitude in approximately 90 s at a concentration of > or = 20 micrograms/ml. They were partially reversed by neostigmine (6 micrograms/ml), and by prolonged washout of the extract. Muscle surface action potentials, recorded with extracellular electrodes, also declined rapidly in amplitude in the presence of the extract. Direct muscle stimulation during inhibition by the extract elicited contractions and action potentials whose magnitudes were similar to control responses. Resting membrane potentials, and the intracellular input impedance of the skeletal muscle cells, were not significantly changed by the alkaloids. These results indicate that the extract has significant neuromuscular blocking activity of a partially or slowly reversible nature. The block appears to be exerted at the postjunctional end plate nicotine receptors, thus offering promise for the identification of novel cholinergic receptor antagonist(s). PMID- 10846628 TI - Possible role of histamine receptors in the central regulation of immune responses. AB - The present study was designed to delineate the role of H1- and H2-histamine receptors in the neuro-immune regulation in rats. The effects of H1- and H2 receptor antagonists on humoral and cell-mediated immune (HI and CMI) responses were investigated after intraperitoneal (i.p.) and intra-cerebroventricular (i.c.v.) administration. HI response was assayed by anti-sheep red blood cell (SRBC) antibody titre in presence and absence of 2-mercaptoethanol (2-ME). The CMI responses were evaluated by delayed type hypersensitivity (DTH) reaction (in vivo), i.e., measurement of footpad thickness, and lymphokine activity such as leucocyte migration inhibition (LMI) test (in vitro). On i.p. administration, both H1- (pheniramine and astemizole) and H2-receptor antagonists (ranitidine and cimetidine) were observed to produce significant enhancement of anti-SRBC antibody response. However, only H2- and not H1-receptor blockers were observed to stimulate CMI response significantly. When administered by icv route, only H2 receptor antagonists caused a statistically significant increase in both HI and CMI responses, while the H1-receptor blockers failed to modify the same. Thus, H2 receptors appear to play a major role in the histaminergic mechanisms involved in immunomodulation both at the level of immunocompetent cells active in the peripheral tissues as well as through the central nervous system structures involved in the central regulation of neuro-immune interaction. PMID- 10846629 TI - Lipid peroxidation and antioxidant enzymes in isoproterenol induced oxidative stress in rat erythrocytes. AB - Isoproterenol, upon oxidation, produces quinones which react with oxygen to produce superoxide anions (O2.-) and H2O2. In the present study, isoproterenol was administered to rats in two doses so as to evaluate its beta adrenergic and toxicological action in terms of lipid peroxidation (LPO) and antioxidant enzymes in erythrocytes. Isoproterenol (30 mg/100 g body wt.) was administered to rats and the animals were followed up to 7 days after administration. Some of these animals were treated with a second dose of isoproterenol 24 h after the first dose and the animals were followed up to 12 h. The result showed increased lipid peroxidation (LPO) and superoxide dismutase (SOD) activity in erythrocytes in response to isoproterenol. Catalase (CAT) activity in erythrocytes decreased with isoproterenol between day 2-7 as compared to control. The second injection of isoproterenol showed increased CAT activity in erythrocytes which decreased at 12 h as compared to control. The erythrocyte GSH content and glutathione-S transferase (GST) activity decreased with isoproterenol treatment as compared to control. However, erythrocyte GSH content as well as GST activity both recovered towards control with time. Elevated serum lactate dehydrogenase (LDH), creatine phosphokinase (CPK) and glutamate oxaloacetate transaminase (GOT) activity was observed after isoproterenol treatment. The results show increased LPO and altered antioxidant system in erythrocytes in response to isoproterenol induced oxidative stress. PMID- 10846630 TI - A study of dyslipidemia and platelet adhesiveness in non-insulin dependent diabetes mellitus. AB - The present study included 50 controls (age 34-64 years) and 50 NIDDM subjects (age 32-72 years) from the diabetic clinic of Government Medical College, Nagpur. It was undertaken with the aim of investigating obesity indices (i.e. body mass index, skin fold thickness, waist hip ratio and % fat in the body); lipid profile (including serum total cholesterol, triglyceride, VLDL, LDL and HDL-cholesterol) levels and platelet adhesiveness in both the groups. On comparison, plasma glucose levels were higher in NIDDM (P > 0.05); obesity indices, cholesterol, triglyceride, VLDL, LDL and platelet adhesiveness index were higher, and HDL levels low in NIDDM group as compared to controls (P < 0.01). Obesity, dyslipidemia and increased platelet adhesiveness are interconnected and make diabetics more susceptible to arterial disease with increased risk of vascular episodes. PMID- 10846631 TI - A study of electroencephalogram in meditators. AB - Electroencephalographic patterns were studied in 30 normal healthy individuals practicing meditation and compared with 10 normal healthy controls not practicing meditation. In this study, we found prominent alpha wave activity and increase it its voltage in meditators as compared to controls. Meditators had significantly more alpha rhythm as compared to control group. Percentage of alpha waves were higher in persons performing meditation with good coherence which suggested good homogenicity, uniformity and increased orderliness of brain. PMID- 10846632 TI - Basal metabolic rate and body composition in elderly Indian males. AB - Aging is associated with a decline in basal metabolic rate (BMR) and total energy expenditure. The extent to which changes in fat free mass (FFM) alone, explains the reduction in BMR is still unresolved. In this study, we documented changes in body composition and evaluated its contribution to measured BMR in young and elderly Indian subjects. We compared 16 healthy elderly male subjects in the age group of 60-75 years with 16 Body Mass Index (kg/m2) matched controls in the age group of 18-25 years. Body composition measurements were determined by using multiple skinfolds and bio-impedence method. BMR measured by indirect calorimetry. The % fat and fat free mass was significantly different between two groups. There were significant difference in BMR between elderly and control group (elderly, 5.91 +/- 0.54 vs control; 7.08 +/- 0.65 MJ/day, P < 0.001), which disappeared when corrected for FFM. This suggests that the age related decrease in the BMR is related to the absolute decrease in the FFM in the elderly without any change in the metabolic activity per kg FFM. PMID- 10846633 TI - Effect of mild irritant on gastric mucosal offensive and defensive factors. AB - The effect of hypotonic medium (Distilled water: DW) and hypertonic saline (HS: 5% NaCl) compared to control normal saline (NS) was studied on gastric ulcer induced by aspirin, 6 h cold restraint stress, ethanol, and pylorus ligation in rats. DW did not afford any protection while HS showed significant ulcer protective effects in all gastric ulcer models studied. The cytoprotective effect of HS seemed to be not only due to its effect on gastric acid secretion but also its effect on mucosal defensive factors like enhanced mucin secretion and decreased cell shedding. As determined by radioimmunoassay, DW did not produce any change in the accumulation of PGE and PGI2, while HS increased them significantly in the human gastric mucosal incubates compared to NS. However, in the incubates of human colonic mucosa, both DW and HS showed a significant increase in PGE with a tendency to increase in PGI2 accumulation. PMID- 10846634 TI - Study of serum lipid and lipoprotein in pre-eclampsia with special reference to parity. AB - A cross sectional study was done to determine serum lipid and lipoprotein cholesterol in pre-eclamptic women in their third trimester, taking normal pregnant women in third trimester as controls. The values were compared in patients of different parity. It was observed that serum triglycerides (TG), cholesterol (Chol), LDL-c, VLDL-c, phospholipids (PL) and total lipids (TL) were significantly raised, while HDL-c, was significantly lower in pre-eclampsia compared to normal pregnancy. TG and VLDL-c were found to be increased significantly with parity. PMID- 10846635 TI - Effect of yoga training on maze learning. AB - The performance in a maze learning task was assessed in adults of either sex (n = 31) before and after 30 days of yoga training and in an age and gender matched control group of subjects who did not receive training in yoga. Subjects were blind folded and used the dominant hand to trace the path in a wooden pencil maze. At each assessment, subjects were given 5 trials, without a gap between them. Performance was based on the time taken to complete the maze and the number of blind alleys taken. The time and error scores of Trial 1 were significantly less after yoga (two-factor ANOVA, Tukey test). Repeating trials significantly decreased time scores at Trial 5 versus Trial 1, for both groups on Day 1 and for the control group on Day 30. Hence the yoga group showed improved performance in maze tracing at retest 30 days later, which may be related to this group being faster learners and also the effect of yoga itself. Yoga training did not influence maze learning, based on the performance in 5 repeat trials. PMID- 10846636 TI - Effect of yogic practices on subjective well being. AB - Forty eight healthy volunteers who participated in the practice of yoga over a period of 4 months were assessed on Subjective Well Being Inventory (SUBI) before and after the course in order to evaluate the effect of practice of yoga on subjective feelings of well-being and quality of life. A significant improvement in 9 of the 11 factors of SUBI was observed at the end of 4 months, in these participants. The paper thus, reiterates the beneficial effects of regular practice of yoga on subjective well being. PMID- 10846637 TI - Effect of selected yogic practices on the management of hypertension. AB - On the basis of medical officers diagnosis, thirty three (N = 33) hypertensives, aged 35-65 years, from Govt. General Hospital, Pondicherry, were examined with four variables viz, systolic and diastolic blood pressure, pulse rate and body weight. The subjects were randomly assigned into three groups. The exp. group-I underwent selected yoga practices, exp. group-II received medical treatment by the physician of the said hospital and the control group did not participate in any of the treatment stimuli. Yoga imparted in the morning and in the evening with 1 hr/session. day-1 for a total period of 11-weeks. Medical treatment comprised drug intake every day for the whole experimental period. The result of pre-post test with ANCOVA revealed that both the treatment stimuli (i.e., yoga and drug) were effective in controlling the variables of hypertension. PMID- 10846638 TI - Effect of sojourn at altitude of 3,500 m on auditory evoked potential in man. AB - Auditory evoked potentials were studied on 27 male subjects to evaluate the effect of sojourn at high altitude (HA). They were initially studied at Delhi (260 m) for their auditory evoked potentials and were then flown to an altitude of 3,500 m. During their stay at HA the same parameter was tested twice, once on the second day of arrival at HA and another on 21st day of stay. Then they were flown back to sea level (SL) and were retested on the third day of return. The results showed a significant delay in the peak latency of wave I of both the right and left ears on the second day of arrival at HA with a persistent delay in wave V even after 3 days of return to SL. The results suggest that HA stress caused a delay in sensory conduction at the cochlear level during the first week of induction, which was normalised during further stay. PMID- 10846639 TI - Comparative study of antioxidant potential of tea with and without additives. AB - Oxidative damage is one of the many mechanism leading to chronic diseases. Therefore interest is growing in the protection afforded by antioxidant nutrients against free radical reactions. More recently, the attention has shifted to polyphenols. Polyphenols are secondary plant metabolites occurring widely in plant food. They possess outstanding antioxidant properties, suggesting a possible protective role in man. Tea (Camellia sinensis) is a widely consumed beverage throughout the world containing polyphenols more than 35% of its dry weight. In the present work we have investigated the effect of tea without milk, tea with milk and lemon tea on the serum lipid peroxidation level (as a parameter of free radical generation). The results show that there were significant decrease in serum lipid peroxidation (Malonaldehyde) level half hour after ingestion of lemon tea and tea without milk which tends to normalize with increase in time. This decrease is much significant in case of lemon tea than tea without milk after half hour or one hour. Hence the interpretation is, tea without milk is a good source of antioxidant and addition of lemon to tea increases its antioxidant potential. PMID- 10846640 TI - A study of cardio-pulmonary efficiency in different categories of runners. AB - Selected short distance runners, middle distance runners and long distance runners were subjected to graded exercise on a treadmill. The maximum aerobic power (VO2 max) and other indices related to oxygen transport system viz. heart rate, ventilation volume, breathing reserve, dyspnoeic index, O2 pulse and RQ were recorded at respective VO2 max work loads, and the values were compared. Long distance runners and middle distance runners showed a significantly higher VO2 max than the short distance runners when VO2 max was expressed per unit of body weight. Among the endurance runners, long distance runners had a significantly lower resting pulse rate as well as the maximum heart rate during work than the middle distance runners. On comparison, Ventilation Volume, Breathing reserve, Dyspnoeic index, O2 pulse and RQ at VO2 max work loads do not differ significantly among different categories of runners. PMID- 10846641 TI - Serum phosphate in acute myocardial infarction. AB - The aim of this study is to measure phosphate levels in AMI, compare and analyse its relation with left ventricular (LV) dysfunction and mortality. Serum phosphate was measured by kinetic assay method in 40 patients with acute myocardial infarction (AMI). Echocardiographic LV function was assessed in all and the patients were followed up for 30 days. Hypophosphatemia (< 2.5 mg/dl) was observed in 27% of AMI patients (11/40). These patients formed group 1 of our study. The rest 73% patients (29/40) with normal phosphate levels formed group 2. Mean Phosphate level in group 1 was 1.96 mg/dl (range 1.2-2.37) and mean ejection fraction (EF) was .35 (range .25-.50, p value < .001). Mean phosphate in group 2 was 3.693 (range 2.6-6.00) and mean EF was .53 (range .38-.65, p value < .001). In hospital mortality of the group 1 was 28% (3/11) while in group 2 was 6.8% (2/29). We conclude hypophosphatemia in AMI is associated with LV dysfunction which results in increased 30 day mortality. PMID- 10846642 TI - Stimulation of iron uptake and Hb synthesis in iron deficient reticulocytes. AB - The effect of inhibitors and intermediates of heme synthesis, inhibitor of globin synthesis, and some iron proteins on in vitro iron uptake and haemoglobin synthesis by reticulocytes of iron deficient subjects was investigated in this study. Lead, INH, ALA, mesoprophyrin, ferritin and albumin substantially increased iron uptake by iron deficient reticulocytes, while cycloheximide and glycine depressed it. The results showed that it is possible to stimulate iron uptake and Hb synthesis in iron deficiency by substances other than iron; the most effective and remarkable of them was ferritin. PMID- 10846643 TI - Antinociceptive effect of gabapentin in rats. PMID- 10846645 TI - Antihypertensive treatment for the neurological patient: a nursing challenge. AB - The neuroscience nurse's role in hypertensive management for patients with neurological injury can be challenging. This is especially true for patients whose cerebral autoregulation is affected by chronic hypertension or a cerebral insult. Hypertensive management involves more than the mere administration of medications. The nurse is responsible for monitoring the effects of drugs, lowering the blood pressure to a safe level and observing for any neurological deficits which may ensue from cerebral hypoperfusion. The nurse must collaboratively be able to determine if the patient's hypertensive episodes are caused by ineffective antihypertensives or a new cerebral insult. Knowledge of hypertension management helps maintain adequate cerebral perfusion and ultimate neurological functioning of the patient. PMID- 10846644 TI - Managing alcohol withdrawal in the acutely ill hospitalized adult. AB - Managing individuals with acute illness who are at high risk for alcohol withdrawal presents multiple challenges to the treatment teams caring for them. Following realization that management of this group was often characterized by severe withdrawal symptoms (delirium tremens, seizures and the need for leather restraints), a Task Force developed protocols to guide care. Its principal goal was to avoid cardiorespiratory and neurologic morbidities associated with severe withdrawal. The first 441 episodes of care treated after protocol implementation are described in this report. There were no instances of oversedation requiring pharmacological reversal or intubation, few individuals suffered seizures outside of the emergency department and the use of leather restraints declined dramatically. Outstanding issues arising from analysis include the necessity of subjecting the symptom severity instrument to rigorous psychometric study and reconsideration of the appropriateness of a symptom-triggered approach in treating this population. Our experience suggests that use of a nonprescriptive approach by educated and motivated nursing and medical staffs can reduce serious morbidity in this at-risk population. PMID- 10846646 TI - Eating patterns in women with multiple sclerosis. AB - For persons with multiple sclerosis (MS), good nutrition has the potential to enhance quality of life and reduce the risk of lifestyle diseases and secondary conditions. The purpose of this study was to describe nutritional intake of women with MS and to determine what factors influenced their intake. Based on 3-day food diary data, participants had an inadequate intake (< 10% lower than recommended) of carbohydrates, dietary fiber, vitamin E, calcium and zinc. In contrast, they consumed saturated fat, protein, vitamin A, vitamin C, folate and iron at higher levels than recommended. Age was significantly related to nutritional intake with older women consuming fewer calories, total fat and saturated fat. Despite evidence supporting the importance of following recommended nutritional guidelines, nutritional intake for participants in this study was not optimum in many important areas. This study supports the need for interventions to promote good nutrition among persons with MS. PMID- 10846647 TI - Emotional distress in critically-injured patients three months after a potentially life-threatening accident. AB - A case-control study was carried out to determine why some critically-injured patients remained emotionally distressed while other critically-injured patients were found to be minimally distressed three months after a potentially life threatening accidental injury. Cognitive processing and meaning theories suggest that psychological adjustment following a traumatizing event depends on the successful integration of the event into current or modified cognitive schema and the restoration of a sense of meaning in life. These cognitive processes may be facilitated by coping strategies that share concerns, mobilize support and reframe disturbing elements. The Global Severity Index was used to identify critically-injured patients as either emotionally distressed or minimally distressed. Bradburn's Psychological Well-Being Scale, Impact of Events Scale, the Meaning-of-Illness Questionnaire and Family Crisis-Oriented Personal Evaluation Scale were used to collect cross-sectional data from 51 critically injured patients during face-to-face interviews 8-12 weeks after the accident. Twenty-seven emotionally distressed patients and 24 minimally distressed patients were compared on the variables of psychological well-being, cognitive processing efforts, restoring a sense of meaning in life and use of coping strategies. Descriptive statistics, ANOVA and MANOVA were used to test for significant differences between the two groups. The findings showed that emotionally distressed patients scored significantly higher on efforts to cognitively process what happened, share concerns and mobilize support. They also had difficulty acknowledging the negative effects of the accident on the self and a poorer sense of psychological well-being. In contrast, minimally distressed patients evidenced a significantly greater ability to acknowledge the negative effects on the self and a higher level of psychological well-being. The potential differences between the two groups in the magnitude of the relationships of meaning, cognitive processing and coping strategies on psychological well-being were assessed by hierarchical regression equations. Psychological well-being of minimally distressed patients was characterized by a significantly higher perception of their capabilities and strengths and by an ability to acknowledge a change in their relations with family and friends. PMID- 10846648 TI - Improving the convenience of home-based interferon beta-1a therapy for multiple sclerosis. AB - Subcutaneous interferon beta-1a (Rebif) therapy has been recognized as a significant advance in the treatment of relapsing-remitting multiple sclerosis (MS). The drug is supplied as a solution in a ready-to-use prefilled syringe. Given the chronic nature of the disease, a convenient and simple drug delivery for self-administration improves patient management. Home-based therapy requires comprehensive knowledge of multiple sclerosis and a training and support program to ensure maximum convenience and independence for the patient. These result in high compliance with therapy and optimum response to treatment. PMID- 10846649 TI - Venous thromboembolism: risk factors in the craniotomy patient population. AB - PURPOSE: The purpose of the study was to determine risk factors for the development of venous thromboembolism in the craniotomy population. Four hundred forty-three patients who under went craniotomy surgery were enrolled in the study. Patients were assessed daily during their postoperative recovery and at the 6-week return postoperative visit for signs and symptoms of thromboembolism. Those not returning for a checkup received a call which asked about the occurrence of signs and symptoms. Chart audits were completed to gather data on risk factors and thromboembolism characteristics. Risk factors included demographic, clinical history, lab, medications, intraoperative and postoperative variables. Deep venous thrombosis and pulmonary embolus symptomatology were also evaluated. Forty-four patients (10%) developed a thrombosis during hospitalization or during the 6 week recovery. The diagnosis was confirmed by duplex scan or venogram. Multiple vessels were the most frequent site. Only 22.7% of the patients had symptoms. Significant risk factors included: presence of leg weakness (p = 0.05), longer LOS (p = 0.0001), more postoperative ICU days (p = 0.001), more preoperative ICU days (p = 0.01), longer recovery time in PACU (p = 0.03), and a longer postoperative period to initiate all activities, ROM, chair, BRP, and ambulating in hall (p = < 0.01). Venous thromboembolism increases morbidity, mortality and health care costs. A risk profile can assist nurses in targeting patients for additional preventive interventions. PMID- 10846650 TI - Diaphragmatic cramp as a possible cause of noncardiac chest pain and referred mandibular pain. AB - The initial assumption that sudden acute chest pain may be of cardiac origin is justifiable, but when this proves not to be the case the patient is left with little explanation of the cause. It is suggested here that diaphragmatic cramp may be a cause of some undiagnosed noncardiac chest pains associated with mandibular referred pain. The phrenic nerve provides both motor and sensory innervation to the diaphragm, while the trigeminal nerve carries sensation from the mandibular teeth. Both nerves originate in separate nuclei close together in the lower medulla. Interconnections between these nuclei and others higher up in the brain may provide one explanation for this problem. PMID- 10846651 TI - [Prevalence of eating disorders in a representative sample of female adolescents from Navarra (Spain)]. AB - BACKGROUND: In spite of the social concern about eating disorders (ED), information of its prevalence in Spain is limited. Further studies in communities are needed to estimate the frequency of this problem in representative samples of the population. SUBJECTS AND METHODS: To estimate the prevalence of anorexia nervosa (AN), bulimia nervosa (BN), and partial syndromes of the "non specified eating disorders" (NOSED) type, a representative sample of the female population of Navarre was selected using a random multistage sampling scheme. A sample of 2862 participants aged 12 to 21 was studied. The screening procedure was performed using the Eating Attitudes Test (EAT) (values over 30 as the cutoff point) and each diagnosis was confirmed using a semi-structured interview performed by a psychiatry according to DSM-IV criteria. RESULTS: The overall prevalence of ED in this female population was 4.1% (95% CI: 3.45-4.95). The disorder specific prevalences being NOSED 3.1% (95% CI: 2.5-3.8), BN 0.8 (95% CI: 0.5-1.2) and AN 0.3 (95% CI: 0.1-0.6). CONCLUSIONS: Our results show a slight lower prevalence than what other Spanish authors have reported, although we found higher prevalences of incomplete syndromes and suggest that a high social burden does exist and primary prevention strategies are needed. PMID- 10846652 TI - [Diagnostic performance of serum thyroglobin and iodide-131 Na scan according to the type of recurrence in differentiated thyroid carcinoma]. AB - BACKGROUND: There is no consensus about the best follow-up protocol in differentiated thyroid carcinoma (DTC). The aim of this study is to evaluate the usefulness and prognostic value of serum thyroglobulin (Tg) and the iodide-131 whole body scan (WBS) in DTC recurrences as a whole and according to the type of recurrence (local recurrence or distant metastases). PATIENTS AND METHODS: Thirty four patients with recurrent DTC recruited in our institution over 15 years, with a minimum 5 years of follow-up and without either distant metastases at time of diagnosis nor detectable anti-Tg antibodies were included in the study. All patients were submitted to total or near-total thyroidectomy and 131INa ablation of postsurgical thyroid remnants. The follow-up included Tg measurement and WBS performed in hypothyroid state. RESULTS: Serum Tg was increased in the 67.4% of the patients and the WBS was positive in the 82.3%. In the isolated local recurrences the sensitivity of WBS was higher than Tg measurement (93.7 vs 43.7%; p < 0.05), but patients with positive Tg had a worse prognostic. By contrast, in patients with distant metastases the sensitivity of Tg was higher than WBS (83.3 vs 58.3%; p = NS). In 14 patients (41.2%) the results of WBS and Tg were in disagreement. In these cases a worse prognosis was observed when Tg was positive and WBS negative. CONCLUSIONS: The sensitivity of Tg and WBS is different depending on the type of recurrence. Therefore, both tests complement each other and it is not recommended to omit one of them in the follow-up of DTC. PMID- 10846653 TI - [Disability from low back pain in Spain]. AB - BACKGROUND: Nonspecific low back pain (LBP) is very common and the economic impact of absenteeism has not been completely established in Spain. MATERIAL AND METHOD: We have valued LBP and the cost of worker's compensation in Spain with the available information between 1993-1997. RESULTS: The mean LBP was 55,388 per year (SD 9,230.1). The transitory LBP disability was 21.9 days, with a range between 19.7 and 24.2 days. The total cost of worker's compensation LBP claims was 11,000 million pesetas/year, and the mean cost by worker was 209,666 pesetas/year. CONCLUSIONS: The worker's compensation LBP claims was higher in Spain that in the European Union. PMID- 10846655 TI - [New indications of permanent cardiac pace maker in the year 2000]. PMID- 10846654 TI - [Analysis criteria's influence on incidence and factors associated tuberculin conversion in health care workers]. AB - BACKGROUND: To know from two different criteria, CDC's and Spanish national consensus (NC), the tuberculin conversion rate and the factors associated to this phenomenon. PATIENTS AND METHODS: A retrospective cohort study was designed over 475 health care workers. Risk factors were identified by using a Cox's proportional hazards model for each criteria. RESULTS: Not being vaccinated with BCG appears to be a risk factor in NC model 5.37 (CI 95%: 2.21-13.00) and does not in CDC. There was a difference of 45% between both incidence density rates. CONCLUSIONS: We did not find a total concordance between the results from the two models. PMID- 10846656 TI - [Recommendations for the medical treatment of severe cranioencephalic trauma. Working group of Intensive Neurology of the Catalan Association of Intensive Health Care (Neuro-ACMI)]. PMID- 10846657 TI - [Current status of the treatment of hepatic metastases of colorectal carcinoma]. PMID- 10846658 TI - [Thrombopoietin: its discovery and clinical perspectives]. PMID- 10846659 TI - [Inhaled glucocorticoids and their effects on growth in children]. PMID- 10846660 TI - [Pacemaker malfunction as a cause of intractable hiccup]. PMID- 10846661 TI - [Alpha-hemolytic streptococci as a cause of purpura fulminans]. PMID- 10846662 TI - [Usefulness of bone densitometry in the evaluation of primary hyperparathyroidism]. AB - BACKGROUND: In recent years, the clinical profile of primary hyperparathyroidism (PH) is predominantly characterized by mild or asymptomatic forms. In this context, the effects of the PH on bone metabolism reach a growing importance in the adoption of therapeutic decisions. PATIENTS AND METHODS: 116 patients with PH were studied, 95 women (25 premenopausal, 70 postmenopausal) and 21 males. In all cases parathyroidectomy criteria were evaluated. Bone mineral density (BMD) was determined in 71 patients in lumbar spine (LS) and femoral neck by dual-X-Ray absorptiometry (DXA) and the influence of this measurement in the decision of surgical treatment was analyzed. RESULTS: The patients with PH showed a significant reduction of BMD at all sites (p < 0.001) and 71.8% met osteoporosis densitometric criteria. The most frequent parathyroidectomy criteria was the presence of specific clinical manifestations (51.7%) followed by decrease of the BMD in LS (49.3%). The probability of meeting criteria for surgical treatment was significantly higher in patients in which BMD was measured (odds ratio: 3.09 [1.03-9.22]; p = 0.036). CONCLUSIONS: In its current presentation, PH presents a significant decrease of bone mass. The systematic performance of bone densitometry has a decisive influence in its appropriate management. PMID- 10846663 TI - [Orthostatic hypotension in the aged and its association with antihypertensive treatment]. AB - BACKGROUND: To assess the prevalence of orthostatic hypotension (OH) in an elderly population and to evaluate the influence of hypertension control and of the type of antihypertensive drug used. METHODS: 2,700 elderly patients (> 65 years) from the EPICARDIAN study were included. Blood pressure (BP) was measured twice with the patient seated and once again 1 to 3 minutes afterwards with the patient standing. Hypertension is defined as BP > or = 140/90 mmHg or if the patient received antihypertensive treatment. Orthostatic hypotension was defined as a fall of SBP > or = 20 mmHg and/or DBP > or = 10 mmHg 1 to 3 minutes after the postural change. RESULTS: 71.6% of the patients were classified as hypertensive and 57.1% were receiving pharmacological treatment. The overall prevalence of OH was 6.8% among hypertensive patients, 8.1% and 3.4% in normotensives patients (p < 0.05). No significant difference was found in the prevalence of OH between those receiving (7.7%) or not (8.6%) antihypertensive medication. No association was found either between the prevalence of OH and the type or number of antihypertensive drugs used. 17.6% of the patients reached hypertension optimal control (SBP < 140 and DBP < 90 mmHg), the prevalence of OH in these patients was 5.3%, among the uncontrolled the prevalence was 8.9% (p < 0.05). The adjusted OR was 1.91 (1.1-3.3) for the prevalence of OH in the uncontrolled patients versus the controlled. CONCLUSIONS: The prevalence of OH in the elderly is not very high, being greater among hypertensive than normotensive patients. No association was found between the prevalence of OH and the number of drugs received. Reaching hypertension control reduces the prevalence of OH. PMID- 10846664 TI - [Incidence of tuberculosis in Spain: results of the Multicenter Project of Tuberculosis Research (MPTR). Work Group of the MPTR]. AB - BACKGROUND: The objective of this project was to gain a better understanding of the incidence of all forms of tuberculosis and to describe the clinical epidemiological characteristics of tuberculosis cases in Spain. METHODS: After active search through 14 registers, we identified cases of tuberculosis within 13 Spanish Autonomous Communities with populations that represented almost 67% of the total population of Spain. The study period was between May 1996-April 1997. RESULTS: The global incidence of all forms of tuberculosis was 38.51 cases per 100,000 inhabitants with some variations between the communities (range: 16.20/100,000-70.75/100,000). A positive smear in the sputum was discovered for 36% of all cases and 49.3% of the cases of pulmonary or miliary tuberculosis. This represented a incidence rate of 13.83 per 100,000 inhabitants (range 4.55/100,000-28.10/100,000). Men presented higher rates of incidence than women (52.70/100,000 versus 24.87/100,000). Concerning age groups, 25-34 years and 75 or more years were the most affected (61.35/100,000 and 59.35/100,000, respectively)). The most frequent sites of the disease were pulmonary, pleural and lymphatic (69.7%, 11.2% and 8.8% respectively) followed by disseminated forms (6.6%). Tuberculosis patients infected with HIV represented 17.7% of the study (range: 7%-27.2%) resulting in a incidence rate of AIDS/tuberculosis of 6.80 per 100,000 inhabitants (range: 2.33/100,000-8.23/100,000). CONCLUSIONS: The results of the Multicentric Project for Tuberculosis Research (MPTR) confirm the importance of tuberculosis in Spain and shows an important interaction between tuberculosis and HIV. PMID- 10846665 TI - [Football, television and emergency services]. AB - BACKGROUND: To know the influence of televised football on the use of emergency department (ED). PATIENTS AND METHODS: We assessed the number, demographic characteristics and acuity of patients attended during the broadcast of football matches played by FC Barcelona during Champions' League (n = 12), and they were compared with days without televised football (n = 12). RESULTS: Televised football was associated with a decrease in visits to ED (-18%; p = 0.002). Such a decrease was observed for all ED units, but only for traumatology unit reached statistical significance (-28%; p = 0.006). Decay of ED visits were mainly due to a decrease of low-acuity consults (-30%; p = 0.04). CONCLUSION: There is a significant decrease on ED use associated with televised football. PMID- 10846666 TI - [Bone densitometry, 2000]. PMID- 10846667 TI - [Epidemiology of eating disorders]. PMID- 10846668 TI - [Results of the RALES trial: good news for patients with heart failure and ... for health administrations]. PMID- 10846669 TI - [Peripartum cardiomyopathy]. PMID- 10846670 TI - [Waldenstrom's disease with osteolytic lesions, renal failure and amyloidosis]. PMID- 10846671 TI - [Pyomyositis caused by Nocardia asteroides in a patient with kidney transplant]. PMID- 10846672 TI - [Central nervous system involvement in tuberculosis]. PMID- 10846673 TI - [Prevalence and factors associated to the presence of fatty liver in apparently healthy adult men]. AB - BACKGROUND: Alcohol intake in one of the factors associated with fatty liver, although its contribution as well as other factors have not been completely established. Therefore the aim of this study was to assess the prevalence and associated factors for fatty liver diagnosed by ultrasonography. SUBJECTS AND METHODS: 1,801 presumably healthy male workers (age range 18-60 years). A complete physical and laboratory investigations, including HBsAg and anti-HCV antibodies, a detailed interview on alcohol intake, and an abdominal ultrasound examination were performed in all cases. Diagnosis of fatty liver was based on defined ultrasonographic criteria. RESULTS: Eighty eight cases were excluded because of the HBsAg or anti-HCV positivity or incomplete ultrasonography. Among the remaining 1,713 cases, 236 (13.8%; 12.2-15.4) had fatty liver. Logistic regression analysis disclosed age (RR: 1.04; CI 95%; 1.03-1.05), ethanol intake > 40 g/d (2.19; 1.81-2.65), gamma-glutamyl-transferase > 40 U/l (3.51; 2.95-4.18), body mass index > 30 (3.87; 3.22-4.66) and glycemia > 120 mg/dl (2.69; 1.85-3.90) as the risk factors for fatty liver. Fatty liver was present in 8.8% of cases who did not have obesity, diabetes or hypercholesterolemia. When the subjects with obesity, hyperglycemia or hypercholesterolemia were excluded, regression analysis confirmed age, ethanol intake and gamma-glutamyl-transferase as independent factors associated with fatty liver. CONCLUSIONS: Age, alcohol intake, obesity, and increased serum levels of glucose, cholesterol and gammaglutamyl transferase are the main factors associated with fatty liver in presumably healthy adult men. PMID- 10846674 TI - [Neuropsychological pattern of cognitive impairment in Alzheimer type dementia and vascular dementia]. AB - BACKGROUND: Cummings et al, 1987, hypothesized that Alzheimer dementia type (ADT) patients would present poorer performances than vascular dementia (VD) patients on the neuropsychological tests that mainly involve cortical neurological structures, and that VD would perform worse on tests that mainly involve subcortical structures. The main purpose of the research was to identify a cognitive impairment pattern that discriminates the type of dementia in the initial stages. METHOD: Two groups of patients, one affected by mild ADT (n = 30) and the other by multi-infarct dementia (VD, n = 30) were given a neuropsychological battery. The battery was composed by Temporal Orientation (Benton et al.); Vocabulary, Similarities, Digits, Coding and Kohs (WAIS) of Wechsler; the Colour-Form Test of Weigh; the Trail Making Test (A and B) (Halstead-Reitan Neuropsychological Battery); Tapping (McQuarrie); Logical Memory, Visual Memory and Paired Association (Wechsler Memory Scale) of Wechsler; Delayed Memory (Russell). The two groups were similar in age and socio-cultural features. The z-score and its statistical significance on the Mann-Whitney test were made and we performed an exploratory discriminant analysis to the classification. RESULTS: In general, results were poorer in the ADT group. But we detected no significant differences in the tests, although some test (Immediate Visual Memory and Kohs' Blocks) almost reached significance. The discriminant analysis reached a classification of the 67% of the subjects into the ADT group and the 70% of the subjects into the VD group. CONCLUSIONS: In the initial stages of dementia it is difficult to differentiate between a cortical pattern of cognitive impairment in ADT and a subcortical pattern in VD, a distinction that the other researchers have reported. When complex tests were used the performance depended on the coordination of multiple related systems. These findings are in agreement with the holistic models of higher mental functions. PMID- 10846675 TI - [Usefulness of Helicobacter pylori antigen detection in stools in the diagnosis of infection and confirming eradication after treatment]. AB - BACKGROUND: A new immunoassay to detect H. pylori antigen in stool (HpSA) has been developed. We started this study to know the sensitivity and specificity of this test as diagnostic tool of H. pylori infection and eradication control. PATIENTS AND METHODS: Forty patients were recruited to study H. pylori infection. At endoscopy, biopsy samples were taken for culture, histology and urease test. Stool specimens were tested by HpSA and serum sample for serology. Patients were defined as H. pylori positive if histology, urease test or culture were positive. Forty-two patients treated with omeprazole based triple therapy were screened 8 weeks after treatment for eradication control using urea breath test, HpSa and serology. Patients with UBT negative were defined as eradicated. RESULTS: As diagnostic tool: 34 out of 40 patients were infected (85%). HpSA was positive in 31 out of 34 patients, and achieve 3 false negative and 1 false positive (sensitivity: 91%, specificity: 84%). IgG anti-H. pylori was positive in 31 out of 34 infected patients, with 3 false positive and 3 false negative (sensitivity 91%, specificity 50%). In eradication control (n = 42), HpSA was negative in 33/38 successfully eradicated patients, and positive in all four non-eradicated patients. Five patients eradicated showed positive HpSA (specificity 87%, positive predictive value: 87%, negative predictive value: 100%). IgG serology was negative in only a third of eradicated patients. CONCLUSIONS: The stool assay was an accurate tool for diagnosis of H. pylori infection and eradication control. PMID- 10846676 TI - [Fatty liver]. PMID- 10846677 TI - [Cortical and subcortical dementia: where is the difference?]. PMID- 10846678 TI - [Antitumor immunology in the year 2000 and the new immunosuppressive therapy]. PMID- 10846679 TI - [Deep venous thrombosis: present and future]. PMID- 10846680 TI - [Myasthenia gravis, thymoma and intestinal pseudoobstruction]. PMID- 10846681 TI - [Hepatitis and nevirapine]. PMID- 10846682 TI - [Unrelated donor bone marrow transplantation in a girl with thalassemia major]. PMID- 10846683 TI - [Angioedema and urticarial reaction induced by valsartan]. PMID- 10846684 TI - [Undiagnosed diabetes and impaired glucose metabolism on high risk Spanish population. The IGT study]. AB - BACKGROUND: To estimate the prevalence of undiagnosed diabetes and impaired glucose metabolism on high risk Spanish population, contrasting phenotypic features according to both sets of criteria, World Health Organization (WHO) and American Diabetes Association (ADA). SUBJECTS AND METHODS: Cross-sectional, multicentre (10 health-care centres, 230,000 inhabitants), selective risk-factor study. WHO diagnoses (normal, impaired glucose tolerance [IGT] and diabetes) were based on the 2 h plasma glucose (2hPG) following a 75 g oral glucose tolerance test (OGTT) and ADA diagnoses according the fasting plasma glucose (FPG) (normal, impaired FPG and diabetes). Prevalence, diagnostic overlap index and 22 clinical and biological parameters were compared. RESULTS: The study included 580 subjects (330 women), mean age 58.1 years and body mass index 31.2 kg/m2, 292 (50.3%) with only one risk factor and 288 (49.7%) with two or more risk factors. WHO diagnoses were: 291 (50.2%) normal glucose tolerance (95% CI: 46.2-54.2%), 157 (27.1%) IGT (23.5-30.7%) and 132 (22.7%) diabetes (19.3-26.1%). ADA diagnoses were: 355 (61.2%) normal FPG (57.2-65.2%), 146 (25.2%) IFG (21.7-28.7%) and 79 (13.6%) diabetes (10.9-16.3%). The prevalence of diabetes decreased 9.1% (from -11.4 to 6.8%). The diagnostic overlap was 33.5% for diabetes and 19.3% for impaired glucose homeostasis (IGT-FPG). Omitting the OGTT half the diabetic subjects with 2hPG > or = 200 mg/dl (11.1 mM) but FPG < 126 mg/dl (7 mM) would remain undiagnosed. By changing screening criteria (FPG instead of 2hPG) the risk related to traditional factors such as age or a first-degree relative with diabetes would reduce. CONCLUSIONS: Applying ADA criteria, the high prevalence of undiagnosed glucose abnormalities would decrease. These results suggest that it is strongly advisable the continued use of the 2hPG for diabetes screening on high risk Spanish population. PMID- 10846686 TI - [Outcome after spontaneous subarachnoid hemorrhage. Relationship between admission glycemia and clinical status on admission and on hospital discharge]. AB - BACKGROUND: To determine the relationship between the glycemia on hospital admission after subarachnoid haemorrhage (SAH) and neurological score. PATIENTS AND METHODS: On 44 patients we recorded the glycemia value on admission, coma Glasgow scale (CGS) and Hunt and Hess scale grade (HH), on admission, on discharge and six months later. RESULTS: The patients with HH IV-V and GCS between 3 and 8 had glycemia values higher than patients with HH I-III and GCS between 9 and 15. The patients with HH IV-V on discharge had glycemia values higher than patients with HH I-III. CONCLUSIONS: Glycemia value on admission is related with the acute injury severity and represents a prognosis factor in spontaneous subarachnoid haemorrhage outcome. PMID- 10846685 TI - [Comparative analysis of articles published by Spanish authors (1993-1997) in biomedical journals with high impact factor]. AB - BACKGROUND: To identify the Spanish scientific production amongst different areas of clinical knowledge, and to compare it with those of five other European Union countries. METHOD: Review of MEDLINE data base, for the period 1993-1997. Search limited to four journals, selected, for 10 different medical specialties (Cardiology, Endocrinology, Infectious Diseases, Gastroenterology-Hepatology, Haematology, Nephrology, Pneumology, Neurology, Oncology, Rheumatology). Articles published by authors from Germany, France, Italy, The Netherlands, Sweden and Spain. Journals included in the Internal Medicine subject classification were independently analysed. Data were also related with several econometric indexes. RESULTS: A total of 1,763 original articles published by Spanish authors were identified in the journal's sample over the analysed period (2.08 articles per 100 all published articles). Spain contributes to the total achieved by the six European countries analysed with 9.07 articles per 100 published articles. Gastroenterology-Hepatology was the medical specially which has more articles published by Spanish authors (total: 338 articles; 4.15 articles/100 published articles); and Oncology the one with less articles published (1.26 articles/100 published articles). The mean IF value per journal by article is highest for Gastroenterology-Hepatology (4.86 FI/article) and lowest for Pneumology (2.42 FI/article). Spain is the last amongst all six European countries analyzed in Endocrinology, Oncology and Haematology, and second to last in all others except for Gastroenterology-Hepatology (4th place). Mean cost for each article produced by Spanish authors in the analyzed sample was 0.49 US $ according the health expenditures per capita, and 0.07 US $ according the R+D expenditures per capita. Data from the independent analysis of Internal Medicine journals also showed that Gastroenterology and Hepatology is the subspecialty with a higher number of papers published in those journals. CONCLUSIONS: All efforts devoted to improve the quality of Spanish biomedical research, specially in clinical research, had produced positive, but uneven, results, measured by the number and impact factor of original articles published in top ranked biomedical journals. The overall distribution of high impact factor scientific production by specialties is poor when compared to the European Union countries included in the analysis. Those results showed several improvement opportunities. Besides increasing the overall budget for R+D, its is likely that the time has come for backing the highest quality Spanish biomedical research, the one that offers greater and better chances for achieving scientifically valid results, and is published in high impact factor biomedical journals. PMID- 10846687 TI - [Home parenteral nutrition. A six-year combined program (adult and pediatric patients)]. AB - BACKGROUND: To describe the outcomes of an adult-pediatric home parenteral nutrition (HPN) program. PATIENTS AND METHODS: Retrospective protocol between 1993 and 1999. RESULTS: Sixteen adults (average 45.7 years) and eight children (3.1 years) were included in the program. Mean length of parenteral nutrition was 507 (SD: 624) and 155 (SD: 129) days respectively. Total follow-up time was 8,119 days for adults and 1,242 for children. Cancer was the main diagnosis in adults and intractable diarrhea in children. Central venous catheter related infections were the most usual complication (0.63 and 1.2 episodes/patient/year). There were no deceases due to the HPN in the period of study. CONCLUSIONS: HPN is an effective and safe technique, although prevalence and incidence in Spain are low. PMID- 10846688 TI - [Glycemia: multi-faceted analytical magnitude]. PMID- 10846689 TI - [Parenteral nutrition y palliative treatment]. PMID- 10846690 TI - [The role of tumor necrosis factor in the control of fat reserve and obesity]. PMID- 10846691 TI - [Lumbar pain, fever, cutaneous lesions and progressing renal insufficiency in a 44-year-old woman with antecedents of recurrent arteriovenous thrombosis]. PMID- 10846692 TI - [Myocardial ischemia in a female patient treated with donepezil]. PMID- 10846693 TI - [Prevalence of HIV infection and risk behavior among drug users]. PMID- 10846694 TI - [Osteoarticular brucellosis: diagnostic use of immunocapture agglutination assay]. PMID- 10846695 TI - [Effectiveness of omeprazole, clarithromycin and amoxycillin therapy to eradicate Helicobacter pylori in patients with active peptic ulcer. Preliminary results of GENPY study]. AB - BACKGROUND: To compare the effectiveness of two length of the same therapy to eradicate Helicobacter pylori in patients with non complicated active peptic ulcer. PATIENTS AND METHODS: 223 patients visited at primary health centres. A randomised controlled trial was carried out to compare the results of 6 or 7 days of treatment with clarithromycin 500 mg, amoxicillin 1 g and omeprazole 20 mg, b.i.d. All patients received omeprazole for other 14 days. RESULTS: 108 patients were randomised to the 6 days group and 115 to the 7 days one. Both groups were comparable with respect to basic characteristics except for sex: the proportion of men was higher in the 6 days group (p = 0.04). 91.4% of the patients received all the treatment correctly. By intention to treat analysis, eradication rates were 76.9% in the 6 days group and 77.4% in the seven days group (IC 95% of the difference from -10.5 to 11.6). There were not statistically significant differences in eradication rates in relation to age, sex, ulcer location, recurrence or tobacco consumption. CONCLUSIONS: Although eradication rates are lower than expected, in this study there are no differences between 6 or 7 days of therapy. PMID- 10846696 TI - [Prevalence of malnutrition and morphofunctional alterations of the pancreas in asymptomatic chronic alcoholic patients]. AB - BACKGROUND: The aim of this study was to assess the prevalence of morphofunctional alterations of the pancreas and the possible association with the nutritional status of chronic alcoholic subjects in the initial phase of detoxification and without symptoms of pancreopathy. PATIENTS AND METHODS: 105 alcoholic patients with neither diagnosis nor clinical symptoms of pancreopathy were studied, from detoxification units for alcoholics with over 30 days abstinence. The nutritional status was evaluated by means of clinico anthropometric parameters (body mass index, nutritional risk index, degree and type of malnutrition) and biological parameters (albumin, transferrin, and total lymphocyte count). The pancreas was morphologically examined by means of plain X rays, ultrasonography and/or computer tomography, applying the Cambridge criteria; also functionally (Pancreolauryl test and fats in stools). RESULTS: The average (SD) consumption of alcohol was 195 (76) g/day (range 60-450), for 16 (8) years (3-40). Nutritional abnormalities were observed between 12.3% and 28% in the clinico-anthropometric parameters, and between 2% and 32% according to the biological parameters. These abnormalities were only light or moderate. The morphologic study showed abnormalities in 3.5% of the cases. Abnormalities in the exocrine function were observed in 26% of the cases, of which one third showed steathorrhea. A significant association between the nutritional condition and the scores of the tests of exocrine function was observed, especially in those cases with steathorrea. CONCLUSION: A large number of asymptomatic alcoholic subjects present a subclinical pancreopathy and signs of malnutrition. Both facts may be related and justify a pancreatic examination with more sensitive techniques in alcoholic individuals with malnutrition. PMID- 10846697 TI - [Cancer mortality in Spain, 1955-1994]. AB - BACKGROUND: To analyze the mortality from the eight main cancer sites in men and women from Spain between 1955-1994. MATERIAL AND METHODS: Age-standardized mortality rates were computed. RESULTS: In men, an increase in lung cancer mortality as well as in the other 8 sites was registered, except for stomach cancer. In women, breast, ovary, and pancreas cancer mortality continue to increase. CONCLUSIONS: Cancer mortality in Spain has dismal trends. This fact implies the need of strong interventions. PMID- 10846698 TI - [Study of bone mineral density in postmenopausal women with rheumatoid arthritis treated with low dose glucocorticoids]. AB - BACKGROUND: To study bone mineral density (BMD) in a group of postmenopausal women with rheumatoid arthritis (RA) treated with low doses of corticosteroids. PATIENTS AND METHODS: One hundred and eleven patients were included. Mean age (SD) was 63.8 (8.8) years, mean duration of postmenopausal period was 16.4 (10.1) years and the mean disease duration was 12.5 (8.2) years. RESULTS: A significant reduction of lumbar BMD (p < 0.05) and femoral BMD (p < 0.0001) was observed. The prevalence of osteoporosis was of 47%. CONCLUSIONS: The study supports, in the Spanish population, that postmenopausal women with RA treated with low doses of corticosteroids, have low BMD. We consider that the prevalence of osteoporosis in these patients is high. PMID- 10846699 TI - [Osteoporosis in rheumatoid arthritis]. PMID- 10846700 TI - [Hyperimmunoglobulinemia D and periodic fever syndrome. A phenotypical analysis of a Spanish family]. AB - BACKGROUND: Hyperimmunoglobulinemia D and periodic fever syndrome (HIDS) is a disorder diagnosed with low frequency, that produces a very prolonged and recurrent fever with other symptoms and analytical markers of inflammation. Its origin seems to be hereditary with a recessive autosomic pattern, but its pathogenic mechanisms are unclear. The aim of this study is to analyse the clinical characteristics and serum levels of immunoglobulins a Spanish family with HIDS. METHODS: We describe a young woman diagnosed with HIDS and investigate the other memberships of her family (parents and 5 brothers) by clinical interview, physical examination, hematological and biochemical analyses and measurements of IgG, IgA, IgM, IgE, IgD and the kappa/lambda ratio of light chains. Moreover, we also determine the IgD in a control group of 35 healthy blood donors. RESULTS: One male brother of the index case also showed a clinical picture of HIDS. The serum IgD levels were increased (above 100 U/ml) in both and in other two sisters without symptoms and were normal in the rest of the family. With only one exception, all individuals of the control group showed a normal IgD level and this was not associated with sex or age. The other immunoglobulins were normal in the family. In spite of the different treatments tested in the index case, only glucocorticoids aborted her fever attacks. CONCLUSIONS: In HIDS the clinical picture and the high IgD levels are both transmitted with a recessive autosomic pattern, but these are not necessarily associated in the same memberships of the family. Its diagnosis is difficult and there is not effective and long-term safe treatment. PMID- 10846701 TI - [Total quality management and evidence based medicine]. PMID- 10846702 TI - [Waiting time in medicine]. PMID- 10846703 TI - [The syndrome of growth hormone deficiency in adults: current criteria for the diagnosis and treatment]. PMID- 10846704 TI - [Q fever endocarditis. Torpid long-term outcome]. PMID- 10846705 TI - [Erectile dysfunction and proton pump inhibitors]. PMID- 10846706 TI - [Primary prevention of cardiovascular events by acetylsalicylic acid in patients with hypertension in Spain]. PMID- 10846707 TI - [Atypical antipsychotics]. PMID- 10846708 TI - [Current assessment of new/atypical neuroleptic agents]. PMID- 10846709 TI - [New i.e. atypical neuroleptic agents for negative symptoms of schizophrenia: results and methodological problems of evaluation]. AB - The results of controlled studies of the efficacy of the new atypical neuroleptics in treating negative symptoms show that these antipsychotics have a more pronounced effect on negative symptoms in acute schizophrenic patients than the classical neuroleptics. Supplementary complex statistical analyses substantiate that the increased efficacy of the atypical neuroleptics in treating negative symptoms can only partially be explained by indirect effects of better extrapyramidal tolerability, better effects on productive psychotic symptoms, etc. Instead, it is due largely to the stronger direct effect of these atypical neuroleptics. Clinical studies to evaluate their efficacy in chronic schizophrenic patients with stable, predominantly negative symptoms are still mostly lacking. First results support the presumption that atypical neuroleptics have a direct effect. Parallel to the evaluation of the new atypical neuroleptics, important progress has been made in the methodology of clinical studies in this area. PMID- 10846711 TI - [External quality assurance of inpatient treatment in schizophrenia. results of a multicenter study]. AB - Due to legal regulations, external quality assurance is mandatory in Germany. Supported by the German Health Ministry (BMG), we present the results of a multicenter study in four hospitals with different structures on 1042 inpatients with the tracer diagnosis of schizophrenia (ICD 10). We defined disease-specific indicators of structure, process, and outcome quality, developed an assessment instrument, and implemented a feedback system for quality comparison. The resulting quality profiles are useful as a starting point for internal quality management. PMID- 10846710 TI - [Phosphorus 31 magnetic resonance spectroscopy in schizophrenia research. Pathophysiology of cerebral metabolism of high-energy phosphate and membrane phospholipids]. AB - 31Phosphorus nuclear magnetic resonance spectroscopy (31P-MRS) has gained much interest in schizophrenia research in the last years, since it allows noninvasive measurement of high energy phosphates and phospholipids of the human brain in vivo. Thus, several studies have reported cerebral metabolic differences between patients and healthy controls as well as on lateralization effects and influences of epidemiological and psychopathological factors. This review gives a survey of the potential of 31P-MRS in schizophrenia research and summarizes and comments on the results of preceding studies. The discussion covers the reduction of phospholipids in patients in the context of the membrane phospholipid hypotheses, the question of an energetic hypometabolism in schizophrenics, and the influence of neuroleptic medication. PMID- 10846712 TI - [Diminution of hand writing area and D2-dopamine receptor blockade. Results from treatment with typical and atypical neuroleptics]. AB - Beside the typical extrapyramidal motor symptoms such as rigidity, tremor, and dyskinesia, a reduction in handwriting area may occur under neuroleptic therapy. To date, the nature of the relationship between a reduction in handwriting area and striatal D2 dopamine receptor occupancy has remained unclear, and it is not known whether such a reduction also occurs under treatment with atypical neuroleptic drugs. In 23 schizophrenic patients treated with haloperidol, haloperidol decanoate, risperidone, and clozapine, the handwriting are was examined using a planimetric computer program. 123I-iodobenzamide (IBZM) single photon emission tomography (SPET) was used to measure the D2 dopamine receptor occupancy. A statistically significant correlation was found between a reduction in handwriting area and D2 dopamine receptor occupancy (r = 0.86; P < 0.0001). The curve derived from the plotted data resulted in a hyperbolic function. The regression was present regardless of whether the patients were treated with typical or atypical neuroleptic drugs. PMID- 10846713 TI - [Treatment of paraphilia and sexually aggressive impulsive behavior with the LHRH agonist leuprolide acetate]. AB - Up to now there are no published results of therapy of paraphilia (Pedophilia, Sadism) and sexual aggressive impulsiveness with LHRH-(luteinizing hormone releasing hormone) Agonists in the Germanspeaking countries. In this report we describe 11 patients which were treated with the LHRH-Agonist Leuprolide Acetate in a period of 12 months. The patients showed no tendency of sexual aggressive behaviour and reported an evident reduction of penile erection, ejaculation, masturbation, sexual deviant impulsiveness and fantasies. One patient died from suicide. In combination with other treatments LHRH-Agonists seem to be a very promising alternative to cyproterone acetate and its possible carcinogene effects. PMID- 10846714 TI - [Differential diagnosis, course and outcome of postpartum psychoses. A catamnestic investigation]. AB - In a polydiagnostic study, a systematically recruited collective of 34 women with a first-episode postpartum psychosis was reexamined after a period of 6-26 years (averaging 12.6 years) in order to establish lifetime-diagnoses according to ICD 10 and Leonhard's classification, and to determine course and outcome. According to ICD-10, unipolar depressive disorders (32%) and acute polymorphous psychotic disorders (28%) represented the most frequent diagnoses. Applying Leonhard's classification revealed a marked predominance of cycloid psychoses (62%) with the subform of motility psychosis being the most frequent diagnosis (38%). Schizophrenias occurred rarely according to both classifications. Investigating the long-term course, we found in 59% multiphasic disorders. The mean number of episodes per patient was 2.5 (range 2-6) with a mean duration of 9.8 weeks (SD = 5.2). 6 patients (18%) had undergone a monophasic course, in 4 cases (12%) the course was not determinable. 17 women (50%) had 19 further deliveries during the follow-up period. The frequency of relapses in connection with a further delivery was 47%. Administering the Strauss-Carpenter-Outcome-Scale revealed a favourable outcome with a mean value of 14.1 (SD = 2.83) for our total sample. Only 4 patients (12%) had never recovered fully since the onset of the illness. Our findings suggest that cycloid psychoses, in particular motility psychoses, account for the majority of postpartum psychoses, and do not support the hypothesis of a nosological independence of postpartum psychoses. They provide further evidence of a favourable prognosis of severe postpartum psychiatric disorder despite a relatively high rate of non-puerperal and especially puerperal relapses. PMID- 10846715 TI - [Investigation of episode-characteristics of unipolar psychotic depression]. AB - The present study examines episode characteristics in unipolar psychotic depression. From a sample of unipolar endogenous depressed inpatients, patients with a psychotic index episode (n = 19) were compared to nonpsychotic patients (n = 86) with regard to case history, characteristics of the inpatient episode, residual symptoms at discharge from hospital and course of illness up to seven months after discharge. Psychotic depressed patients displayed more severe observer-rated depressive symptoms at admission and were more likely to have attempted suicide prior to admission. In the post-discharge short-term course of depression, these patients showed a more pronounced symptom homogeneity in the extreme ranges, which occurred by stable remissions or by prolonged symptomatology in need of treatment. These findings, together with the observation of higher stabilities of symptom scores in the psychotically depressed, emphasize the prognostic significance of symptomatology at discharge to the post-discharge episode course in these patients. PMID- 10846716 TI - [Valproic acid in prophylaxis of bipolar disorder. A case of valproate-induced encephalopathy]. AB - A 28-year-old patient with a 5-year history of bipolar disorder developed signs of encephalopathy 2 weeks after the addition of valproic acid to his treatment regimen of doxepine, risperidone, and biperidene. The clinical signs were drowsiness, ataxic gait, asterixis, and a generalized epileptic seizure. Discontinuation of valproic acid gradually resulted in complete remission of these symptoms. Valproate encephalopathy has been described mainly in patients receiving anticonvulsant polytherapy. This complication might become more prevalent in psychiatric pharmacotherapy due to the increasing use of valproic acid. PMID- 10846717 TI - [Acute psychiatric symptoms as the initial manifestation of HIV-infection: differential diagnosis, therapy and medico-legal issues]. AB - The prevalence of psychiatric symptoms and disorders in HIV-infected patients is high. The differential diagnosis includes psychoreactive disorders, acute psychiatric symptoms of an HIV-associated encephalopathy, and symptomatic psychotic illnesses due to secondary neurologic manifestations such as opportunistic central nervous infections and intracerebral lymphoma. Clinical aspects and psychopathological findings are not sufficient for differential diagnosis and identification of primarily psychiatric disorders. Secondary neurologic manifestations causing a symptomatic psychosis must be excluded as soon as possible by brain imaging (CT, MRI) and analysis of the cerebrospinal fluid. In emergency situations, however, German law imposes strict regulations, especially in the case of sectioned patients. These medical and medicolegal questions are illustrated by case reports and propositions for an effective strategy are made. PMID- 10846718 TI - [Kufs-disease; a rare cause of early-onset dementia]. AB - The case of a 35-year-old man with progressive dementia from the age of 17 is presented. Clinical examination showed mild extrapyramidal and cerebellar signs and rare myoclonus. Neuropsychological evaluation disclosed severe cognitive deficits. Magnetic resonance imaging (MRI) revealed moderate generalized atrophy with abnormal iron deposition in the basal ganglia. Positron emission tomography (PET) with 18-fluorodeoxyglucose (18-FDG) demonstrated clear temporoparietal hypometabolism. The clinical symptoms and course are typical for the rare adult type of neuronal ceroid lipofuscinoses (Kufs' disease). The diagnosis is supported by the electron microscope detection of an abnormal accumulation of lipid vacuoles and lipofuscin in the eccrine sweat glands and the rectal ganglia cells. PMID- 10846719 TI - [Follow-up study after enteral manganese poisoning: clinical, laboratory and neuroradiological findings]. AB - Manganese intoxication is an unusual, severe form of intoxication. This report deals with a patient now 80 years old who accidentally ingested a solution of potassium permanganate for a period of at least 4 weeks 14 years ago. Since then, the patient suffers from a mild parkinsonian syndrome and distally accentuated polyneuropathies. Psychiatric disorders, especially demential or depressive symptoms, were not observed. Manganese analysis of his hair still shows a clear increase in manganese concentration. The MRI of his brain showed no pathological changes, in particular none of those often described with symmetric signal elevation in T1 in the area of the basal ganglia. In this study, we present clinical, laboratory, and neuroradiological findings. Unusual in this case with a short exposition is the long duration and clinical improvement without L-dopa treatment. PMID- 10846720 TI - [The state of psychiatry in Ukraine. Psychiatric hospitals in Kiev and Shitomir]. AB - The situation of psychiatric patients in the Ukraine is determined by the financial and economic difficulties of this country. In the larger cities, so called polyclinics take care of ambulant psychiatric patients. However, no other social psychiatric institutions exist. Here, the modalities of hospital psychiatric treatment are discussed using the psychiatric hospitals in Kiev and Shitomir as examples. Undertreatment is also a problem, because only a few psychopharmacological drugs are produced in the Ukraine. Import of new psychopharmacological agents is hampered by economical problems. Improving the availability of psychopharmacological drugs and the establishment of a social psychiatric network are major aims for psychiatrists working in Ukraine. PMID- 10846722 TI - [In Process Citation] PMID- 10846721 TI - [Which German psychiatric institution was first in building a psychiatric hospital? Comments on the title picture and text of Nervenarzt 1999, issue 70]. PMID- 10846723 TI - [Adjuvant whole body acupuncture in depression]. PMID- 10846724 TI - [Treatment of dissociative symptoms in borderline personality disorder with naltrexone: supplementary comments]. PMID- 10846725 TI - We want to partner in improving the oral health of children. The Academy and the American Board of Pediatric Dentistry. PMID- 10846727 TI - Erbium YAG laser--a surprising clinical finding. PMID- 10846726 TI - The Wand revisited. PMID- 10846729 TI - Ferric sulfate pulpotomy in primary molars: a retrospective study. AB - PURPOSE: This investigation collected clinical and radiographic data from a retrospective chart review of patients receiving ferric sulfate pulpotomies with a sub-base of zinc oxide eugenol in a clinical practice over a five-year period. METHODS AND MATERIALS: Clinical and radiographic data were available for 242 primary molars in 171 children with follow-up times ranging from 4-57 mos (mean = 19 mos). RESULTS: The overall radiographic success rate ranged from 74-80%. The frequency of normal appearing pulps decreased over time. The most frequently observed pulpal responses were calcific metamorphosis (6-33%) and internal resorption (7-18%). Overall clinical success was 99%. Only 9 of the 242 teeth were extracted due to radiographic and/or clinical failure. A survival analysis demonstrated that the overall cumulative probability of survival remained high over time with a cumulative survival of over 90% after 3 years. CONCLUSIONS: The overall success rates in this study are lower than those reported previously in the literature for ferric sulfate pulpotomies, but are comparable with those reported for 1:5 dilution, 5-min formocresol pulpotomies. PMID- 10846728 TI - Non-nutritive sucking behaviors in preschool children: a longitudinal study. AB - PURPOSE: Prolonged duration of non-nutritive sucking behaviors may have consequences in regard to the developing orofacial structures and occlusion. Little is known as to why some children have prolonged sucking habits beyond the first 2 to 3 years of life. This paper reports on non-nutritive sucking patterns among a large cohort of healthy children from birth to 36 months of age and older, and identifies factors predictive of prolonged non-nutritive sucking habits. METHODS: Over 600 children were followed from birth to at least 36 months of age using mailed questionnaires sent when children reached the ages of 6 weeks, and 3, 6, 9, 12, 16, and 24 months, and then yearly thereafter. Parents answered questions concerning non-nutritive sucking behaviors including use of pacifier and digit sucking. The study categorized children who maintained habits to 36 months of age or older as having prolonged habits, and using multivariate analyses, compared them to children without prolonged habits on various sociodemographic variables. RESULTS: The study found that for over 20% of the children, a non-nutritive sucking habit was prolonged to 36 months of age or older. Factors associated with prolonged sucking habits included older maternal age, higher maternal education level, and having no older siblings. CONCLUSIONS: Identifying factors related to prolonged non-nutritive sucking habits may be important in developing and targeting recommendations regarding such behaviors in an effort to prevent malocclusions that result from prolonged sucking habits. PMID- 10846730 TI - Masticatory function in patients with juvenile rheumatoid arthritis. AB - PURPOSE: Children with Juvenile Rheumatoid Arthritis (JRA) rarely report temporomandibular joint (TMJ) pain, which may be due to pain avoidance mechanisms resulting in compromised masticatory function. This study examined the relationship between self-report measures of pain and dysfunction and measures of chewing performance in 44 JRA children and 34 normal controls. METHODS: The children were divided into three groups: Group 1, JRA children with temporomandibular joint disorder (TMD); Group 2, JRA children without TMD; Group 3, normal control children without TMD. Both visual and analog scales of jaw pain, ability to chew, and quality of life were administered before and after chewing tasks. Children chewed standardized portions of an artificial food for 20 cycles and expectorated the particles into a cup. This process was repeated five times. Median particle size and a broadness of particle distribution index were measured. Also, the number of chewing cycles prior to the child's first swallow for a cube of carrot was recorded. RESULTS: The broadness of particle distribution index was greater for Group 1 (P < 0.001) and Group 2 (P < 0.03) than for Group 3 with no difference in number of chews for carrot mastication among groups. Group 1 reported more pain and dysfunction before the chewing tasks than Groups 2 or 3 (P < 0.05). Interestingly, only Group 3 reported increased pain and decreased ability to chew after chewing tasks (P < 0.02). CONCLUSION: Children with JRA compromise their masticatory function as a pain avoidance mechanism. Such findings may have profound implications with regard to the nutritional status for these children. PMID- 10846731 TI - Dental caries in HIV-infected children versus household peers: two-year findings. AB - PURPOSE: This report will present a two-year comparison of the incidence and baseline prevalence of dental caries found in both the primary and permanent dentition among a cohort of HIV-infected children as compared to household peer control subjects who were not HIV-infected. METHODS: The subjects in this report were from an initial cohort of 171 children (104 HIV positive and 67 HIV negative), who were participants in the Children's Hospital AIDS Program in Newark, New Jersey, from 1993-1995. This two year analysis reports the findings on the children who completed baseline through Year 02 examinations (N = 121), aged 2-15 years old (68 HIV positive, 53 HIV negative). RESULTS: While the DMFS incidence at Year 02 among the 6-11 year old control subjects was 17% higher than that of the HIV-infected cases (2.1 vs. 1.8, respectively) this same incidence was eight-fold higher for the control subjects among the 12-15 year olds (e.g., 8.1 vs. 1.0, respectively). The mean cumulative dmfs score to date for HIV infected cases was higher than for the control subjects for both the 2-5 year olds and the 6-11 year olds, (11.0 vs. 7.0) and (10.0 vs. 4.0, P = .02), respectively. In all three age groups, HIV-infected cases had a greater number of primary teeth and fewer number of permanent teeth than the control subjects (P < .01). CONCLUSION: Given that HIV-infected cases had lower DMFS scores and higher dmfs scores than their household peer controls, the fewer mean number of permanent teeth among the HIV-infected cases suggests that this delayed tooth eruption pattern in permanent teeth contributed to the lower DMFS scores seen in the HIV-infected cases. PMID- 10846732 TI - Oral soft tissue manifestations in HIV-positive vs. HIV-negative children from an inner city population: a two-year observational study. AB - PURPOSE: Data accrued after two years of longitudinal observation of oral soft tissue lesions in a cohort of HIV-infected children and comparisons to a group of uninfected controls is presented. SUBJECTS AND METHODS: One hundred and four HIV positive subjects were enrolled from an inner city pediatric HIV clinic and HIV negative household peers served as control. Oral exams were performed at six month intervals while laboratory data of interest were obtained from the children's medical records. RESULTS: HIV-positive children had significantly more oral soft tissue lesions than their HIV-negative peers. In particular, the prevalence of candidiasis, linear gingival erythema and median rhomboid glossitis were high. However, oral lesions were not good predictors of mortality and only candidiasis was associated with a low CD4 count. CONCLUSIONS: Oral soft tissue lesions were common among HIV-positive children. While candidiasis was correlated with advanced disease, oral lesions were not good predictors of mortality. PMID- 10846733 TI - Reports of pain by children undergoing rapid palatal expansion. AB - PURPOSE: This study described and quantified the prevalence, timing, and intensity of pain during the expansion phase of rapid palatal expansion (RPE) in children and investigated whether pain was related to age, sex, or rate of expansion. METHODS: Ninety-seven children, 38 males and 59 females, between the ages of 5 to 13 years (median 7.7 years) undergoing RPE procedures with the Hyrax, Dentaurum, Newtown, PA, appliance were surveyed. The appliance was expanded with either one or two turns (1/4 mm/turn) per day based on the provider's preference. The child's pain response was measured no more than 5 minutes after each turn for the entire period of expansion using both the Facial Pain Scale and the Color Analog Scale. RESULTS: Ninety-eight percent of the children reported at least some pain during RPE. The highest levels of pain were reported during the first 10 turns with the greatest intensity during the first 6 turns and a steadily decreasing amount of pain thereafter. Pain medication was taken after 7% of the expansion turns in the study with the majority of children taking the medication during the first 6 turns. Forty-eight percent of the children took pain medication at least once during the expansion phase of RPE. There was no difference in either reported pain or use of pain medication based on age, sex, or stage of dentition. During the first 10 turns, children whose rate of expansion was two turns/day were more likely to report pain and take pain medication than children whose rate of expansion was one turn/day, thereafter there were no differences. CONCLUSIONS: The vast majority of children undergoing the active phase of rapid palatal expansion with a Hyrax appliance report pain. The pain generally occurs during the initial phase of expansion and diminishes thereafter, with two turns/day resulting in reports of pain greater than those expanding only once/day. PMID- 10846734 TI - Alternate inexpensive pediatric dental chair. PMID- 10846735 TI - Clinical guidelines for treatment of impalement injuries of the oropharynx in children. PMID- 10846736 TI - Peripheral giant cell granuloma: a potentially aggressive lesion in children. AB - A slowly enlarging gingival mass with a reddish-purple surface is observed in a school-age boy. The lesion was first noted 3 months ago during a routine oral examination but recently it has increased in size and interferes with eating. A periapical radiograph demonstrated focal loss of the alveolar crestal bone in the mandibular incisor region. The diagnosis of peripheral giant cell granuloma, a benign reactive gingival lesion, is confirmed by histopathologic examination. Early detection and excision of this hyperplastic nodule is important to minimize potential dentoalveolar complications. PMID- 10846737 TI - Identification of Candida dubliniensis in a study of HIV-seropositive pediatric dental patients. AB - PURPOSE: The combination of an immature immune system and suppressed cellular immunity in children with HIV infections provides optimal conditions for rapid disease progression. As a result, pediatric AIDS has become a major epidemiological challenge. Oral fungal colonization remains one of the most common opportunistic infections observed in both adult and pediatric HIV infected patients. Although Candida albicans is the most frequently isolated opportunistic fungal species, a recently characterized Candida species, C. dubliniensis, has gained considerable attention due to its almost exclusive association with HIV seropositive individuals. The purpose of this study was to prospectively screen for the presence of C. dubliniensis among pediatric HIV+ patients. METHODS: Oral samples taken from twenty-seven children were cultured for the presence of yeast. All positive yeast isolates obtained were screened for the presence of C. dubliniensis by use of tests for germ tube and chlamydospore production, detection of inability to grow at 45 degrees C, by colony color on CHROMagar Candida medium, coaggregation with Fusobacterium nucleatum ATCC 49256 and by the results of sugar assimilation testing with the API 20C AUX yeast identification system. RESULTS: Among the 27 patients tested, 3 patients were found to harbor C. dubliniensis, one of which also grew C. glabrata; 12 patients were colonized with C. albicans, while the remaining 12 patients were negative for yeast. Identification of the three C. dubliniensis isolates was genetically confirmed by electrophoretic karyotyping. All three C. dubliniensis isolates were found to be susceptible to fluconazole (MIC < or = 0.25 ug/ml). CONCLUSIONS: These results confirm the presence of this novel species in a dental pediatric HIV seropositive population and support the need for further investigation into the prevalence and pathogenesis of C. dubliniensis. PMID- 10846738 TI - Nitric oxide in plants. To NO or not to NO. AB - The current knowledge on the occurrence and activity of NO in plants is reviewed. The multiplicity of nitrogen monoxide species and implications for differentiated reactivity are indicated. Possible sources of NO are evaluated, and the evidence for the presence of nitric oxide synthase in plants is summarised. The regulatory role of NO. in plant development and in plant interactions with microorganisms, involving an interplay with other molecules, like ethylene or reactive oxygen species is demonstrated. Finally, some other suggestions on potential functions of NO. in plants are indicated. PMID- 10846739 TI - Purification and characterisation of vanadium haloperoxidases from the brown alga Pelvetia canaliculata. AB - Two enzymes characterised as iodoperoxidases (PcI and PcII), with vanadium dependent activity, have been purified from the brown alga Pelvetia canaliculata (L.) Decne et Thur. (Fucaceae, Phaeophyceae), collected in the Northern Portuguese coast, at Viana do Castelo. The relative molecular masses were 166 kDa for PcI and 416 kDa for PcII, as determined by gel filtration. SDS-PAGE shows that PcI has just one band corresponding to a subunit of 66 kDa, while PcII shows four bands (66, 72, 157 and 280 kDa). The following kinetic parameters have been determined from a steady-state analysis of the oxidation of iodide by H2O2: PcI, pHopt = 6.0, KM(I-) = 2.1 mM, KM(H2O2) = 110 microM, Ki(I-) = 127 mM; and PcII, pHopt = 6.5, KM(I-) = 2.4 mM, KM(H2O2) = 20 microM and Ki(I-) = 69 mM. These iodoperoxidases are thermostable, as also observed for vanadium bromo- and chloroperoxidases. PMID- 10846740 TI - The configuration of methyl jasmonate affects paclitaxel and baccatin III production in Taxus cells. AB - All stereoisomers of methyl jasmonate (MJA) were prepared, and their effects on cell yield and promotion of paclitaxel (Taxol) and baccatin III production investigated in cell suspension cultures of Taxus media. (3R,7S)-MJA showed the strongest cell growth inhibition, followed by (3R,7R)-MJA. In contrast, (3S,7R)- and (3S,7S)-MJA had very low inhibitory effects, indicating that this inhibition depends largely on the (3R)-configuration. In terms of the promotion of paclitaxel and baccatin III production, (3R,7R)-MJA had the highest activity. Although it showed considerable activity at low concentration, at higher concentrations the activity was decreased due to strong inhibition of cell growth. Interestingly, paclitaxel and baccatin III contents increased even at a high (3S,7R)-MJA concentration, whereas the other isomers had the opposite effects. These findings are interpreted to suggest that the optimum configuration is (3R,7R), the (3R)-configuration not being indispensable, and that the (7R) configuration is suitable for the promotion of paclitaxel and baccatin III production. PMID- 10846741 TI - Differential expression of four sweet potato peroxidase genes in response to abscisic acid and ethephon. AB - Expression of four peroxidase (POD) genes, three anionic PODs (swpa1, swpa2 and swpa3), and one neutral POD (swpn1) isolated from suspension cultures of sweet potato (Ipomoea batatas) were analyzed by measuring the accumulation of transcripts in suspension cultured cells and leaves of sweet potato in response to the stress-related plant hormones abscisic acid (ABA) and ethephon (an ethylene generating chemical). The four genes responded differently to ABA (0.1 mM) and ethephon (0.1 mM) in cultured cells and leaves. In suspension cultures, ABA reduced the expression levels of swpa1, swpa2, and swpn1, but did not affect the level of swpa3. Ethephon strongly increased expression levels of swpa3 and swpn1, and slightly increased the level of swpa1. The expression level of swpa2 was reduced. Expression levels in intact leaves, however, were significantly changed by this treatment. Expression of the swpa1 and swpa2 genes was induced 15 min after ABA treatment, followed by a decrease to a basal level after 3 h. A strong re-expression occurred after 12 h. Expression of the swpa3 and swpn1 genes occurred from 3 to 24 h after treatment. All four genes were differentially expressed 12 h after ethephon treatment. The swpa2 gene was strongly expressed immediately after ethephon treatment. The results indicate that each POD gene is differentially regulated by ABA and ethylene in whole plants and in cultured cells in vitro. PMID- 10846742 TI - Microbial hydroxylation/functionalization of terpenoid synthons derived from communic acids. AB - Incubation of a communic acid-derived synthon with Cunninghamella elegans quantitatively affords 1 beta, 3 beta- and 7 beta- monohydroxylated derivatives. PMID- 10846743 TI - Insecticidal piperidine alkaloid from Microcos paniculata stem bark. AB - The stem bark of Microcos paniculata contained a new alkaloid, N-Methyl-6 beta (deca-1',3',5'-trienyl)-3 beta-methoxy-2 beta-methylpiperidine, which showed good insecticidal activity against Aedes aegypti second instar larvae. PMID- 10846744 TI - The elicitin secreted by Phytophthora palmivora, a rubber tree pathogen. AB - Palmivorein, a new member of the elicitin family, was purified from the culture filtrate of Phytophthora palmivora isolated from the rubber tree, Hevea brasiliensis. The elicitin was obtained by ammonium sulfate precipitation and further purified using ion-exchange and gel filtration. The molecular weight, isoelectric point, amino acid composition and N-terminal sequences of this molecule are reported and compared to other known elicitins. Palmivorein, as determined by SDS-PAGE, is a small protein of M(r) ca. 10,000. It is classified as an alpha-elicitin according to its acidic pI and the valine residue at position 13. Like other elicitins, the P. palmivora elicitin causes tissue necrosis on tested tobacco leaves. It also causes severe wilting and necrosis of Hevea tissue, and leaves of the susceptible rubber clone (with respect to P. palmivora) are much more sensitive to this elicitin than those that are resistant. PMID- 10846745 TI - Biotransformation of cadinane sesquiterpenes by Beauveria bassiana ATCC 7159. AB - Incubation of cadina-4,10(15)-dien-3-one with Beauveria bassiana ATCC 7159 has resulted in the production of nine novel sesquiterpenes. These metabolites were identified as (4S)-cadin-10(15)-en-3-one, (4S)-3 alpha-hydroxycadin-10(15)-ene, (4R)-3 alpha-hydroxycadin-10(15)-ene, (4S)-3 beta-hydroxycadin-10(15)-ene, (4S)-3 beta-hydroxycadina-10(15),12(14)-diene, (4S)-13-hydroxycadin-10(15)-en-3-one, (4S)-12-hydroxycadin-10(15)-en-3-one, (4R)-3 beta, 14-dihydroxycadin-10(15)-ene and 3 alpha-hydroxycadina-4,10(15)-diene. The allylic alcohol 3 alpha hydroxycadina-4,10(15)-diene was also biotransformed to afford cadina-4,10(15) dien-3-one, (4S)-cadin-10(15)-en-3-one and (4S)-12-hydroxycadin-10(15)-en-3-one. The insecticidal potential and phytotoxicity of the isolated metabolites have been evaluated. PMID- 10846746 TI - Naturally durable heartwood: evidence for a proposed dual defensive function of the extractives. AB - We previously proposed that extractives in highly durable heartwood may protect wood against fungal colonization and subsequent degradation by dual mechanisms: the extractives have some fungicidal activity and are also free radical scavengers (antioxidants). In short-term laboratory decay tests using two different wood species and decay fungi, the antioxidant 2,6-dimethyl-di-tert butyl-4-methylphenol (BHT) alone had little or no preservative effect. In contrast, the combination of BHT with different organic commercial biocides always showed an increase in efficacy compared to the organic biocide alone. Consequently, we conclude that the combination of a commercial antioxidant and biocide is synergistic. This implies that extractives may protect wood by more than simply being fungicidal. PMID- 10846747 TI - Antimicrobial intermediates of the general phenylpropanoid and lignin specific pathways. AB - The minimum inhibitory concentration (MIC) of the major intermediates of the general phenylpropanoid and lignin specific pathways of plants were determined employing a range of yeasts and bacteria. Of the three main classes of compounds tested, the hydroxycinnamaldehydes were the most effective, possessing notable antifungal and antibacterial activity. Determination of the minimum killing concentration (MKC) of the hydroxycinnamaldehydes revealed MIC/MKC ratios suggesting these compounds to be fungicidal, but not bactericidal, in their mode of action. In contrast, the hydroxycinnamic acids and hydroxycinnamyl alcohols possessed little antimicrobial activity, with the exception of the hydroxycinnamic acids, which were antibacterial. PMID- 10846748 TI - The cyanobacterium Nodularia PCC 7804, of freshwater origin, produces [L Har2]nodularin. AB - The cyanobacterium Nodularia PCC 7804, an axenic, non gas-vacuolate strain from a freshwater source, produces several metabolites with cyanobacterial hepatotoxin characteristics. The most abundant is a cyclic pentapeptide, [L Homoarginine2]nodularin. [L-Har2]nodularin is of similar toxicity, in terms of bioassay in vivo, and the inhibition of protein phosphatase-1 in vitro to nodularin, which was present in lesser amounts in the cultures. PMID- 10846749 TI - Oligostilbenoids in stem bark of Vatica rassak. AB - Three resveratrol oligomers, vaticanols. A, B and C, as well as three known stilbenoids, resveratrol, piceid and epsilon-viniferin were isolated from the stem bark of Vatica rassak (Dipterocarpaceae). Their structures were determined by the analysis of NMR spectral data including the application of 2D methods. PMID- 10846750 TI - African Cucurbita pepo L.: properties of seed and variability in fatty acid composition of seed oil. AB - Pumpkin (Cucurbita pepo) seeds are used locally in Eritrea to treat tapeworm. Seeds were found to be rich in oil (approximately 35%), protein (38%), alpha tocoferols (3 mg/100 g) and carbohydrate content (approximately 37%). The physico chemical properties and fatty acid composition of the seed oil were examined. The four dominant fatty acids found are: palmitic C16:0 (13.3%), stearic C18:0 (8.0%), oleic C18:1 (29.0%) and linoleic C18:2 (47.0%). The oil contains an appreciable amount of unsaturated fatty acids (78.0%) and found to be a rich source of linoleic acid (47.0%). Within the three localities of the study, variations exist in seed properties and the fatty acid composition of the oil. PMID- 10846751 TI - Triterpene saponins from Randia formosa. AB - Seven new triterpenoid saponins, randiasaponins I (1), II (2), III (3), IV (4), V (5), VI (6) and VII (7) as well as two known ones, ilexoside XXVII (8) and ilexoside XXXVII (9), were isolated from the methanolic extract of the leaves of Randia formosa. The structures of the new saponins were established as 3-O-alpha L-arabinopyranosyl-3 beta,19 alpha,23-trihydroxyursa-12,20(30)-dien-28-oic acid 28-beta-D-glucopyranosyl ester (1), 3-O-beta-D-glucopyranosyl-(1-->3)-alpha-L arabinopyranosyl rotundic acid (2), 3-O-beta-D-glucopyranosyl-(1-->3)-alpha-L arabinopyranosyl pomolic acid 28-beta-D-glucopyranosyl ester (3), 3-O-alpha-L rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranosyl pomolic acid 28-beta-D glucopyranosyl ester (4), 3-O-alpha-L-rhamnopyranosyl-(1-->2)-alpha-L arabinopyranosyl siaresinolic acid 28-beta-D-glucopyranosyl ester (5), 3-O-alpha L-arabinopyranosyl ilexosapogenin A 28-beta-D-glucopyranosyl ester (6), and 3-O beta-D-glucopyranosyl ilexosapogenin A 28-beta-D-glucopyranosyl ester (7), based on spectral and chemical evidence. Besides the saponins, two common flavonoids kaempferol 3-O-rutinoside and rutin were also isolated. PMID- 10846752 TI - Contribution of each caffeoyl residue of the pigment molecule of gentiodelphin to blue color development. AB - To clarify the function of each caffeoyl residue in the diacylated anthocyanin gentiodelphin, a pigment from the blue flower of Gentiana makinoi, two mono deacyl derivatives were compared for both color development and stability. In neutral solution, 3,5-di-O-beta-D-glucopyranosyl-3'-O-(6-O-caffeoyl-beta-D- glucopyranosyl)delphinidin was both bluer and more stable than 3,3'-di-O-beta-D glucopyranosyl-5-O-(6-O-caffeoyl-beta-D- glucopyranosyl)delphinidin. Conformational analysis of each derivative under acidic conditions revealed only the 3'-O-caffeoylglucopyranosyl derivative to demonstrate intramolecular stacking. Additionally, the acyl residue in the B-ring contributed more to blue color development than that in the A-ring. PMID- 10846753 TI - Alkaloids and triterpenoids from Ammocharis coranica (Amaryllidaceae). AB - The bulbs of Ammocharis coranica yielded eight alkaloids: lycorine, acetylcaranine and crinamine, which have been reported previously from A. coranica, 1-O-acetyllycorine, hippadine, 6 alpha-hydroxypowelline and hamayne, which have been reported from other members of the Amaryllidaceae, 1-O-acetyl-9-O demethylpuviine, which has not been described previously, and the known cycloartane compounds: 24-methylenecycloartan-3 beta-ol, cycloeucalenol, cycloeucalenone and also 24-methylenepollinastanone, which has not been described previously. PMID- 10846754 TI - Gum arabic glycoprotein contains glycomodules of both extensin and arabinogalactan-glycoproteins. AB - Gum arabic glycoprotein (GAGP) is a large molecular weight, hydroxyproline-rich arabinogalactan-protein (AGP) component of gum arabic. GAGP has a simple, highly biased amino acid composition indicating a repetitive polypeptide backbone. Previous work (Qi, W., Fong, C., Lamport, D.T.A., 1991. Plant Physiology 96, 848), suggested small (approximately 11 residue) repetitive peptide motifs each with three Hyp-arabinoside attachment sites and a single Hyp-arabinogalactan polysaccharide attachment site. We tested that hypothesis by sequence analysis of the GAGP polypeptide after HF-deglycosylation. A family of closely related peptides confirmed the presence of a repetitive 19-residue consensus motif. However, the motif: Ser-Hyp-Hyp-Hyp-Thr-Leu-Ser-Hyp-Ser- Hyp-Thr-Hyp-Thr-Hyp-Hyp Leu-Gly-Pro-His, was about twice the size anticipated. Thus, judging by Hyp glycoside profiles of GAGP, the consensus motif contained six Hyp-arabinosides rather than three and two Hyp-polysaccharides rather than one. We inferred the glycosylation sites based on the Hyp contiguity hypothesis which predicts arabinosides on contiguous Hyp residues and arabinogalactan polysaccharides on clustered non-contiguous Hyp residues, i.e. the GAGP motif would consist of arabinosylated contiguous Hyp blocks flanking two central Hyp-polysaccharides. We predict this rigidifies the glycoprotein, enhances the overall symmetry of the glycopeptide motif, and may explain some of the remarkable properties of gum arabic. PMID- 10846755 TI - New isoflavones from Ceiba pentandra. AB - Two new isoflavones, pentandrin (1) and pentandrin glucoside (2), were isolated from the stem barks of Ceiba pentandra along with beta-sitosterol and its 3-O beta-D-glucopyranoside, which was isolated for the first time from this plant. The structures of these compounds were elucidated with the help of spectroscopic techniques, while the structure of 1 was unambiguously confirmed by single crystal X-ray diffraction studies. PMID- 10846756 TI - Functional MR imaging of the craniocervical junction. Correlation with alar ligaments and occipito-atlantoaxial joint morphology: a study in 50 asymptomatic subjects. AB - PURPOSE: Whiplash injuries are frequent in industrialized countries. The acute and chronic symptoms following such injuries are incompletely understood and objective clinical or imaging findings are rare. Several authors have suspected that rear end collisions occurring when the head is rotated may result in tears of the alar ligaments. There has been experimental proof that a torn alar ligament increases the rotation of the C0/C1 and C1/C2 segments to the contralateral side. Functional cross sectional imaging has therefore been proposed to diagnose injuries of the alar ligaments. So far, published data on normal ranges of rotation in an asymptomatic population have been sparse. The aim of this study was to determine by MR imaging the normal range of rotation in the first three cervical segments and their relation to the morphology of the alar ligaments and the occipito-atlantoaxial joints. MATERIAL AND METHODS: Functional MR imaging of the craniocervical junction in maximum active left and right head rotation was performed in 50 healthy volunteers with a mean age of 29.8 years (31 men, 19 women, range 19-47 years). Measurements were independently performed by two musculoskeletal radiologists to assess interobserver error. The results were correlated with gender and age, with morphological findings in the occipito atlantoaxial joints (i.e. joint symmetry, joint effusions, dens position), and with the form, course and symmetry of the alar ligaments. RESULTS: The mean range of rotation for the C0/C1 joint was 2.7 degrees (standard deviation [SD] 3.3 degrees)/3.3 degrees (SD 3.6 degrees) (right/left) and at the C1/C2 level 38 degrees (SD 6.5 degrees)/37.8 degrees (SD 6.4 degrees). The mean differences in left/right rotation were: C0/C1 3.5 degrees (SD 2.8 degrees) and C1/C2 6.3 degrees (SD 4.4 degrees). No correlation was found between segmental rotation and morphological characteristics of the craniocervical joints or ligament structures. CONCLUSION: There is wide variation of segmental motion in the upper cervical spine. Differences in right-to-left rotation are frequently encountered in an asymptomatic population. Therefore, these measurements are unsuitable for indirect diagnosis of soft tissue lesions after whiplash injury and should not be used as a basis for treatment guidelines. PMID- 10846757 TI - Continuous monitoring of liver oxygenation with near infrared spectroscopy during naso-gastric tube feeding in neonates. AB - Near infrared spectroscopy (NIRS) is a non-invasive method of estimating the haemoglobin concentration changes in certain tissues. It is frequently used to monitor oxygenation of the brain in neonates. At present it is not clear whether near infrared spectroscopy of other organs (e.g. the liver as a corresponding site in the splanchnic region, which reacts very sensitively to haemodynamic instability) provides reliable values on their tissue oxygenation. The aim of the study was to test near infrared spectroscopy by measuring known physiologic changes in tissue oxygenation of the liver in newborn infants during and after feeding via a naso-gastric tube. The test-retest variability of such measurements was also determined. On 28 occasions in 25 infants we measured the tissue oxygenation index (TOI) of the liver and the brain continuously before, during and 30 minutes after feeding via a gastric tube. Simultaneously we measured arterial oxygen saturation (SaO2), heart rate (HR) and mean arterial blood pressure (MAP). In 10 other newborn infants we performed a test-retest analysis of the liver tissue oxygenation index to estimate the variability in repeated intra-individual measurements. The tissue oxygenation index of the liver increased significantly from 56.7 +/- 7.5% before to 60.3 +/- 5.6% after feeding (p < 0.005), and remained unchanged for the next 30 minutes. The tissue oxygenation index of the brain (62.1 +/- 9.7%), SaO2 (94.4 +/- 7.1%), heart rate (145 +/- 17.3 min-1) and mean arterial blood pressure (52.8 +/- 10.2 mm Hg) did not change significantly. The test-retest variability for intra-individual measurements was 2.7 +/- 2.1%. After bolus feeding the tissue oxygenation index of the liver increased as expected. This indicates that near infrared spectroscopy is suitable for monitoring changes in tissue oxygenation of the liver in newborn infants. PMID- 10846758 TI - [Acute myocardial infarction: time delay from onset of pain to hospital presentation and thrombolysis]. AB - BACKGROUND: The efficacy of thrombolytic therapy in patients with myocardial infarction depends largely on the delay to initiation of treatment. METHODS: From February 1996 to May 1997 we prospectively studied 175 patients hospitalised for acute chest pain and suspected coronary heart disease. We determined the delays between onset of pain and calling for medical help, hospital arrival and thrombolysis. RESULTS: The median delay until hospital presentation was 3.1 hours, with no difference between male and female or younger and older patients. 35% of the patients arrived at the hospital within 2 hours, 38% within 2-6 hours and 27% after more than 6 hours. Most of the delay was caused by the patients (median 2.0 hours). Emergency doctors contributed 35 minutes, and transport to the hospital took 31 minutes. 99 patients (57%) were diagnosed with acute myocardial infarction. 53 (54%) fulfilled the criteria for thrombolysis and were so treated. The median overall delay from onset of pain to thrombolysis was 2.9 hours. 77% received treatment within 6 hours, and 23% within 6-12 hours after onset of pain. The median delay inside the hospital (door to needle) was 40 minutes. CONCLUSIONS: 75% of patients with acute chest pain arrived at the hospital within 6 hours of onset of symptoms. Most of the delay was caused by the patients themselves, a factor which needs to be improved by better education. Moreover, optimising in-hospital processes should further shorten the delay to thrombolysis. PMID- 10846759 TI - [Non-surgical closure of patent ductus arteriosus: acute and long-term results]. AB - INTRODUCTION: Closure of a patent ductus arteriosus (PDA) even with small shunt volumes in asymptomatic patients is recommended because of the risk of endocarditis (1.5% per year) and the potential development of congestive heart failure or pulmonary hypertension. METHODS: 16 patients (9 adults [5 men/4 women] and 7 children [3 boys/4 girls]) underwent transcatheter closure of a PDA. The intervention was performed using the Rashkind umbrella in 12 cases, the Amplatzer duct occluder in 2 cases, a coil in 2 cases, and the Sideris self-adjusting device in one case. Echocardiographic controls were carried out one day, 6 months, 12 months and 24 months after the intervention. RESULTS: PDA closure was complete immediately following the intervention in 9 of 16 patients (56%). At follow-up (mean 19 months) complete closure was observed in 14 of 16 patients (87%). All residual shunts were trivial. The complete closure rate in adults was 100% and in children 71%. Complications such as device embolism, endarteritis and haemolysis did not occur. CONCLUSION: Non-surgical closure of a PDA is a safe and effective technique. PMID- 10846760 TI - [Liver changes in telangiectasia hemorrhagica hereditaria (Osler disease)]. PMID- 10846761 TI - [Stage-based management of bronchial carcinoma: the surgeon's point of view]. AB - Surgery is the treatment of choice for non-small cell lung carcinoma (NSCLC) stage I and II. However, adjuvant or neoadjuvant chemotherapy may prove to be of benefit for stages IB and II. Treatment of non-small cell lung carcinoma stage IIIA remains controversial. Recently, neoadjuvant chemotherapy has shown excellent results in patients with metastases to mediastinal lymph nodes discovered at mediastinoscopy and, therefore, seems to be the treatment of choice for these patients. Patients with non-small cell lung carcinoma stage IIIB and IV may benefit from surgery in rare and extremely well selected cases. PMID- 10846762 TI - [Recent development in the treatment of lung cancer: from the point of view of the medical oncologist]. AB - After a short introduction to recent developments in the field of tobacco control, the non-surgical treatment of small cell and non-small cell lung cancer is summarised with special reference to practical aspects. The promising results obtained with newer cytostatic agents are reviewed. In small cell lung cancer, modern chemotherapy prolongs survival five-fold to an average of one year and 5 10% of all patients achieve long-term disease-free survival. In non-small cell lung cancer combination chemotherapy prolongs median survival and improves the quality of life. Newer cytostatic agents such as the taxanes, vinorelbine and gemcitabine are highly active. These agents are currently incorporated into new, highly active combination chemotherapies. Whether these newer polychemotherapies will bring a significant improvement awaits the results of future randomised trials. PMID- 10846763 TI - [The mucociliary system of the lung--role of surfactants]. AB - Many pollution particles enter the organism via the lung. In the lung, on a surface of 140 m2, the blood is separated from the air by a tissue barrier of only 1/1000 mm. The conducting airways (trachea, bronchi, bronchioli) are a very effective aerodynamic filter for inhaled particles. The mucociliary transport system functions like a self-cleaning mechanism within the filter. Inhaled particles and particles deposited in the lungs play a crucial aetiological and therapeutic role. The discussion in health policy on the relationship between the increase in air pollution and lung damage is of great importance at the present time. Epidemiological studies of recent years have shown very clearly that there is a correlation between morbidity and mortality as a consequence of respiratory and cardiogenic problems and the concentration of PM10 particles in ambient air. So far, however, this correlation has not been explained. The intrathoracic airways are coated by a respiratory epithelium. This has an irregular coating of viscous liquid, consisting of a low viscous sol phase and a high viscous gel phase. It seems, however, that those phases are not clearly distinguishable. The gel phase is moved towards the pharynx by the metachronal ciliary beat transporting the particles out of the lungs. Furthermore, at the air-liquid interface, there exists a continuous surfactant film which reduces the surface tension as is the case in the alveoli. When particles are deposited on the airway wall, that is, on the surfactant film, they are wetted by surface forces and displaced into the liquid phases. Thus, the surfaces of the particles are probably changed by the surfactant or by surfactant components. Many of these particles are transported in the liquid (gel phase) towards the pharynx (mucociliary transport), whereas some of them remain in close association with the epithelium (sol phase). Such particles remain in the airways for days or even weeks. They are either phagocytised by macrophages and carried off via the airways or taken up by dendritic cells and transported into the tissue from where they reach the lymph nodes via lymph drainage and are presented to the T lymphocytes. The displacement of particles into the liquid phases, caused by the surfactant, can be considered as the initial step in a complex cascade of defence processes in the lungs. The surface of the particles is probably modified by surfactant or surfactant components. These modified particles may be directed to that clearance pathway which is most beneficial for our health, that is, out of the lungs or into the lymphatic glands, where an immune reaction can be triggered. We therefore consider surfactant to be a primary immune barrier. PMID- 10846764 TI - Function and ultrastructure of cilia in primary ciliary dyskinesia. AB - Primary ciliary dyskinesia (PCD) is a heterogeneous disease with impaired mucociliary transport leading to upper and lower respiratory disorders, hearing impairment and male infertility. Primary ciliary dyskinesia can only be diagnosed by clinical features together with functional and structural analysis of the cilia. To prevent bronchiectasis with marked reduced quality of life, early diagnosis is essential. For this purpose we compared our experience over 10 years with the literature. Our concept consists of a thorough interdisciplinary examination of the patient to rule out other underlying pathologies such as allergy, cystic fibrosis, immune deficiencies, and alpha-1-antitrypsin deficiency on the basis of their clinical features. Thereafter, mucosal biopsies from 27 patients were investigated. In 10 patients (37%) primary ciliary dyskinesia was diagnosed with the help of functional and ultrastructural analysis. 9 patients displayed no or impaired ciliary motility and a high percentage showed ultrastructural defects such as dynein arm deficiency, radial spoke defects and translocation of peripheral microtubular doublets with absent central microtubules. We suggest that investigation of mucosal biopsies for primary ciliary dyskinesia diagnosis without clinical preselection is indicated in patients with "situs inversus" suffering from chronic and/or recurrent airway infections, in patients with neonatal respiratory distress syndrome of "unknown" cause (i.e. after ruling out the other well known causes) with "situs inversus". PMID- 10846765 TI - [Clinical features of primary ciliary dyskinesia]. AB - Primary ciliary dyskinesia is a recessively inherited group of disorders with abnormal ciliary activity leading to disturbed mucociliary clearance. Clinical manifestations as early as the first year of life are recurrent rhinitis, otitis media, sinusitis and lower respiratory tract infections. Another typical presentation is situs inversus. Biopsy of the ciliated mucosa in the nose or bronchi is required for study by vital microscopy and electronmicroscopy to confirm the diagnosis. Early diagnosis is important for initiation of rigorous treatment involving physiotherapy, inhalation with beta 2-mimetics and prompt antibiotic treatment to prevent irreversible damage such as bronchiectasis. Compared with cystic fibrosis the prognosis is better, because older children can compensate the absent mucociliary clearance with always-functioning cough clearance. PMID- 10846766 TI - [Mucous clearing respiratory-physiotherapy in pediatric pneumology]. AB - Chest physiotherapy for mucus clearance may be seen as a therapeutic application of respiratory physiology; the basic mechanical principles do not differ between the treatment of paediatric and adult patients. However, children present with a developmentally defined, age-specific situation regarding respiratory structure and function that calls for a specialised therapeutic approach. Inspiration dilates bronchi and thus brings air behind obstructing mucus plugs. Newborns and infants have the special problem of an overly compliant, instable thorax and an easily obstructed upper airway; hence they require special help with lung volume management. Forced expirations mobilise and transport secretions by expiratory airflow in combination with a wave of moving choke points. Early in life, low stability of airways threatens to hamper this mechanism by complete closure; this calls for special therapeutic assistance. Positioning regionally modifies ventilatory excursions and is thus used to target gas-liquid pumping. However, lateral decubitus position does not affect ventilation in children in the same way as in adults. Disease-inflicted changes of structure and function call for further modification of the therapeutic approach. Finally, the psychology of therapist-patient interaction differs substantially between different age groups. PMID- 10846767 TI - [A rare case of deep bone and pelvic vein thrombosis or: when the lower cavity vein fails]. PMID- 10846768 TI - Is less intensive fetal surveillance of dichorionic twin gestations justified? PMID- 10846769 TI - Cervical morphology in pregnancy, bacterial vaginosis and the risk of preterm delivery. PMID- 10846770 TI - Fetuses and infants with congenital urinary system anomalies: correlation between prenatal ultrasound and postmortem findings. AB - OBJECTIVE: Detection of congenital urinary system anomalies is an important part of the prenatal ultrasound examination. The present study compares prenatal ultrasonographic findings and postmortem examinations of fetuses and infants with renal and urinary tract anomalies. DESIGN: Criteria for inclusion were an ultrasound examination at the National Center for Fetal Medicine (Trondheim, Norway) and autopsy performed during the period 1985-94. Results from the ultrasound examination and autopsy regarding urinary system anomalies were categorized according to the degree of concordance. RESULTS: Urinary system anomalies were found in 112 (27%) of 408 fetuses with congenital anomalies. The renal and/or urinary tract anomaly was the principal reason for induced abortion or cause of death in 50 cases (45%). In 97 (87%) of the 112 cases there was full agreement between the ultrasound observations and the autopsy findings. In five cases the autopsy revealed minor findings not mentioned in the ultrasound report. The main diagnosis was thus correct in 102 cases (91%). In four cases major autopsy findings had not been found by ultrasound examination; in another four, none of the autopsy findings were suspected by ultrasound, and in two, minor ultrasound findings were not confirmed at autopsy. CONCLUSIONS: The accordance between ultrasound diagnoses and postmortem examinations proved to be satisfactory. The close co-operation between ultrasonographers and perinatal pathologists is mutually beneficial. In addition to complementing prenatal diagnosis, postmortem examination is of vital importance for the quality control of ultrasonography in fetal diagnosis and plays an important role in genetic counseling. PMID- 10846771 TI - Striking findings concerning the variability in the measurement of the fetal renal collecting system. AB - OBJECTIVE: Using strict thresholds of normality, investigators have reported associations between fetal pyelectasis and aneuploidy, obstructive uropathy, postnatal pyeloplasty and vesicoureteral reflux. Although evidence continues to mount regarding the importance of fetal pyelectasis, little is known of the variability of collecting system measurements. To investigate the short-term variability and its relationship to bladder dilatation, the following study was conducted. DESIGN: During May and June 1996, 20 mid- and late-trimester patients with varying degrees of fetal pyelectasis were recruited into this study. Each consented to undergo periodic (every 15 min for 2 h) ultrasound measurements of the fetal renal collecting systems and bladder. To account for anatomic and technical variability in measurement, we used both the sum of the transverse and anterior-posterior (AP) measurements of the collecting systems, and AP measurement alone for comparison. The variability of the collecting system measurements was assessed based upon gestational age, magnitude of pyelectasis and fetal bladder size. RESULTS: We investigated 38 renal units in 20 patients (eight in the second trimester and 12 in the third). The mean(SE) AP diameter of all kidneys was 5.89(2.49) mm (range 15 (2-17) mm), and the sum of transverse and AP renal collecting system diameters was 13.91(5.73) mm (range 26 (4-31) mm). These data were normally distributed. When assessing the variability in individual kidneys over time, we found the mean variation (minimum to maximum) for the sum of the AP and transverse measurement to be 7.61(4.26) mm and for the AP measurement alone to be 3.80(2.49) mm. No relationship was found between variability of dilatation, magnitude of dilatation or fetal bladder size. Cyclic dilatation of the fetal bladder was observed in all cases. The mean time from maximal to minimal dilatation was 20 min (1.34 observations; range 12-30 min). CONCLUSIONS: We found the size of the fetal renal collecting system to be highly variable over the course of a 2-h period. Seventy per cent of cases (14 of 20) had both normal (< 4 mm) and abnormal values (> or = 4 mm) during the 2-h study period. Significant caution should be used when considering the implications of renal collecting system dilatation based upon a single AP measurement. PMID- 10846772 TI - Fetal compensatory renal hypertrophy with a unilateral functioning kidney. AB - OBJECTIVE: To evaluate the possibility of fetal compensatory renal hypertrophy with a unilateral functioning kidney. METHODS: A nomogram of renal length was established from 254 normal fetuses. Renal length was also obtained in 14 fetuses with unilateral renal agenesis and 22 fetuses with a unilateral multicystic kidney. RESULTS: Compensatory renal hypertrophy, defined as a renal length > 95th percentile for gestational age, occurred in 16 of 36 cases (44.4%). CONCLUSIONS: Compensatory renal hypertrophy is detectable in utero and may occur as early as 22 weeks' gestation. PMID- 10846773 TI - Functional urinary tract obstruction developing in fetuses with isolated gastroschisis. AB - OBJECTIVE: To evaluate the frequency and natural history of urinary tract abnormalities developing in fetuses presenting with initially isolated gastroschisis. METHODS: Serial ultrasounds were performed prospectively on fetuses identified by our prenatal diagnosis program as having a gastroschisis. When abnormalities in the urinary tract were identified prenatally, newborns were evaluated by a pediatric urologist. RESULTS: Over a 1-year period four out of 12 fetuses with gastroschisis developed deformations of the urinary tract. In three fetuses the bladder herniated through the abdominal wall defect. Two also had upper tract dilatation. A fourth fetus developed bilateral hydronephrosis with a normally situated bladder. Once the gastroschisis was repaired none of the newborns had evidence of structural obstruction of the urinary tract, however, hydronephrosis with or without reflux persisted for several months. CONCLUSIONS: Deformations of the fetal urinary tract can develop secondary to gastroschisis. They do not appear to represent separate malformations and evaluation with fetal karyotyping may not be indicated. When hydronephrosis is present ongoing urologic evaluation of the neonate is indicated. PMID- 10846774 TI - Combined ultrasound biometry, serum markers and age for Down syndrome risk estimation. AB - OBJECTIVE: To compare Down syndrome screening efficiency of the standard serum triple analyte screen to that of a four-component screen consisting of ultrasound biometry and serum markers in the second trimester. METHODS: The Down syndrome screening efficiency of the triple screen, i.e. alpha-fetoprotein (AFP), unconjugated estriol (E3), hCG and maternal age, was compared with the four marker algorithm, i.e. humerus length, nuchal thickness, AFP and hCG plus maternal age. A quadrivariate Gaussian algorithm was used to calculate individual Down syndrome odds. Receiver operating characteristic (ROC) curves plotting sensitivity against false-positive rate were constructed for each algorithm and the areas under the curves were compared to determine which was superior. Sensitivity and false-positive rates at different Down syndrome risk thresholds were also compared. RESULTS: There were 46 cases of Down syndrome (1.9%) with 2391 normal singleton pregnancies in a referral population in which triple screen, fetal biometry and karyotype had been done. The gestational age range for the study was 14-24 completed weeks. The median maternal age for the study group was 35.0 years (14.0-46.0 years). The areas (SE) under the ROC curves were 0.75(0.04) and 0.93(0.02) for the standard triple and the four-marker screen, respectively (P < 0.001). At a 10% false-positive rate, detection was 45.7% for the triple and 80.4% for the four-marker screen. CONCLUSIONS: A new algorithm combining humerus length and nuchal thickness measurement with serum AFP, hCG and maternal age substantially improved Down syndrome screening efficiency compared with the traditional triple screen. The model appears promising and should be evaluated in an independent data set. PMID- 10846775 TI - Middle cerebral artery peak systolic velocity in the prediction of fetal anemia. AB - OBJECTIVE: The fetal middle cerebral artery peak systolic velocity (MCA PSV) has been suggested as a potential test to predict the fetal hematocrit level. We tested the hypothesis that a low fetal hematocrit is associated with an increase in MCA PSV in a prospective study of normal and alloimmunized pregnancies. METHODS: Fetal hematocrit and MCA PSV were obtained in 26 alloimmunized fetuses, immediately before their first fetal blood transfusions between 15 and 35 weeks. Results were compared with the MCA PSVs from 170 control fetuses not at risk of alloimmune anemia between 13 and 37 weeks. RESULTS: In control fetuses, PSV varied with gestation (PSV = 0.56 - 0.032 GA + 0.00086 GA2, where GA is gestational age; R2 = 0.41). The correlation between PSV and hematocrit Z scores (Pearson correlation coefficient r = -0.69) was highly significant (P = 0.0001). Using a PSV > 1 SD, the sensitivity of the test in predicting a fetal hematocrit < 2 SD below the mean was only 64% but the specificity was 100%. However, the sensitivity of the test in predicting a fetal hematocrit < 3 SD and < 4 SD rose to 73% and 83%, while the specificity was still good (93% and 80% respectively). CONCLUSIONS: MCA PSV and fetal hematocrit are highly significantly correlated. The sensitivity of the test was good and the high positive predictive value indicates that the presence of a raised PSV (defined as > 1 SD) is a strong indicator of fetal anemia. In conclusion, MCA PSV is a useful test in clinical practice for the detection of fetal anemia. PMID- 10846776 TI - Fetal cerebral blood flow redistribution in late gestation: identification of compromise in small fetuses with normal umbilical artery Doppler. AB - OBJECTIVE: To evaluate the role of middle cerebral artery Doppler in small fetuses during the late third trimester. DESIGN: Prospective observational study of structurally normal fetuses with an estimated fetal weight < 5th percentile for gestation. Perinatal outcome was determined using a structured datasheet sent to each referring obstetrician. SUBJECTS: Structurally normal fetuses at 35 or more weeks of gestation referred during a 2-year period to the fetal growth clinic of a regional fetal medicine unit in North London. Fetuses with aneuploidy and/or major structural abnormalities were excluded. METHODS: Umbilical artery and middle cerebral artery (MCA) Doppler waveforms were recorded and considered abnormal if above 95th or below 5th percentiles, respectively. Amniotic fluid was considered reduced if the maximum vertical cord-free pool was < 2 cm. The placenta was considered mature if the Grannum grade was II or III. The head circumference (HC)/abdominal circumference (AC) ratio was considered abnormal if > 95th percentile for gestation. Fetal growth, amniotic fluid, biophysical profile score and umbilical artery Doppler were used to advise the referring obstetrician about fetal well-being and he/she independently decided both the timing and mode of delivery. RESULTS: Forty-seven fetuses fulfilled the entry criteria. Thirty-four (72%) demonstrated normal umbilical artery Doppler waveforms. Sixteen (34%) demonstrated middle cerebral artery redistribution, of which nine (56%) had normal umbilical artery Doppler waveforms. MCA blood flow redistribution was associated with an increased incidence of cesarean delivery and need for neonatal admission. Of all gray-scale parameters, an elevated HC/AC ratio has the strongest association with MCA blood flow redistribution (15/16 vs. 1/31; P < 0.01). CONCLUSIONS: MCA Doppler may be a useful tool to assess the health of small fetuses in the late third trimester. Redistribution may occur in the presence of normal umbilical artery Doppler and should be suspected when the HC/AC ratio is elevated. PMID- 10846777 TI - Blood flow velocity waveforms of the fetal pulmonary artery and the ductus arteriosus: reference ranges from 13 weeks to term. AB - OBJECTIVE: To establish reference ranges for blood flow velocity waveforms of the fetal main pulmonary artery and the ductus arteriosus during the second and third trimesters of pregnancy. METHODS: A prospective cross-sectional echocardiographic study was performed in 222 normal fetuses from 13 to 41 weeks of gestation using high resolution/color Doppler ultrasound equipment. RESULTS: Gestational age specific reference ranges are given for peak velocity across the pulmonary valve, velocities in the ductus arteriosus, peak systolic velocity in the ductus arteriosus-to-peak velocity across the pulmonary valve ratio, ductal systolic velocity-to-diastolic velocity ratio, ductal resistance index and ductal pulsatility index. In the ductus arteriosus, increasing fetal heart rate was significantly associated with increasing end-diastolic velocity and decreasing systolic-to-diastolic ratio, decreasing resistance index, and decreasing pulsatility index. CONCLUSIONS: Based on a prospective study in more than 200 normal fetuses, the data provide gestational age specific reference ranges for blood flow velocity waveforms of the fetal pulmonary artery and the ductus arteriosus. The reference ranges may be helpful in prenatal diagnosis of cardiac malformations and ductal obstruction from 13 to 41 weeks of gestation. PMID- 10846778 TI - Reference ranges for two-dimensional echocardiographic examination of the fetal ductus arteriosus. AB - OBJECTIVES: To establish reference ranges for 2D-echocardiographic examination of the fetal ductus arteriosus and its relationship to the main pulmonary artery and the aorta. METHODS: A prospective cross-sectional echocardiographic study was performed in 222 normal fetuses from 13 to 41 weeks of gestation using high resolution/color Doppler ultrasound equipment. RESULTS: Gestational age-specific reference ranges are given for the diameter of the pulmonary valve anulus, diameter of the ductus arteriosus at its beginning, middle, and end, ductal length, ductal diameter-to-pulmonary valve anulus diameter ratio, and the spatial relationship of the ductus arteriosus to the main pulmonary artery and to the aorta. CONCLUSIONS: The presented data derived from a study group of 222 normal fetuses provide in-vivo insights into the morphology of the ductus arteriosus and its relationship to the adjacent vessels. The reference ranges may be helpful in prenatal diagnosis of cardiac malformations and abnormalities of the ductus arteriosus, such as obstruction or aneurysm from 13 to 41 weeks of gestation. PMID- 10846779 TI - Fetal aortic arch measurements between 14 and 38 weeks' gestation: in-utero ultrasonographic study. AB - OBJECTIVE: To establish in-utero reference ranges for fetal transverse aortic arch diameter (TAD) and distal aortic isthmus diameter (DAID) using high resolution ultrasound techniques. DESIGN: A prospective, cross-sectional study was performed on 125 normal singleton pregnancies between 14 and 38 weeks' gestation. Transverse and diameter and DAID were measured by transvaginal ultrasonography until 17 weeks' gestation, and by abdominal ultrasound between 18 and 38 weeks' gestation. RESULTS: Transverse arch diameter as a function of gestational age was expressed by the regression equation TAD = -1.17 + 0.169 X GA, and DAID = -1.39 + 0.189 X GA; TAD and DAID are transverse aortic and distal aortic isthmus diameters expressed in millimeters and GA is gestational age in weeks. The correlation r = 0.924 and 0.938 was found to be highly statistically significant (P < 0.001) for TAD and DAID. The normal mean of TAD and DAID per week and the 95% prediction limits were also defined. CONCLUSIONS: The normative data established by us may be helpful in the prenatal diagnosis of congenital heart defects, including abnormal growth of the aortic arch. PMID- 10846780 TI - Prenatal diagnosis of abnormalities of the fetal venous system. AB - OBJECTIVE: To present our experience in the prenatal diagnosis of anomalies of fetal veins using high-resolution color Doppler ultrasound. DESIGN: An observational study of 16 fetuses with abnormalities of the umbilical, portal, hepatic and caval venous system being diagnosed at the Division of Prenatal Diagnosis and Therapy (Bonn, Germany) over the past 5 years. The abnormality of the venous system, the underlying embryologic disorder and the outcome of the pregnancy are presented and compared with the literature. RESULTS: In group A, eight fetuses had an abnormal course of the umbilical vein with a patent (n = 3) or absent (n = 5) ductus venosus. No portal veins and absent or abnormal hepatic veins were visualized by color Doppler sonography. Six fetuses (75%) did not have an associated malformation and have survived. Two pregnancies with fetal hydrops due to a small heart and to Turner's syndrome were terminated or ended in fetal demise. In group B, seven of eight fetuses with an abnormal caval system had a situs ambiguus or an atrial isomerism. A cardiac defect was detected in six cases (86%). These six pregnancies ended in four terminations of pregnancy and two infant deaths due to the severity of the congenital cardiac defect. One child with a normal heart and a child with an isolated abnormal course of the lower inferior vena cava are developing well. CONCLUSIONS: In a targeted fetal scan the course of the umbilical vein, ductus venosus, the portal and hepatic veins and inferior vena cava should be carefully examined using color Doppler. Any suspicious finding should be followed by a detailed assessment of the specificity of this abnormality taking into consideration the embryologic development of the fetal venous system together with the associated malformations. PMID- 10846781 TI - Morphology assessed by transvaginal ultrasonography differs in patients in preterm labor with vs. without bacterial vaginosis. AB - OBJECTIVE: To determine whether cervical morphology in preterm labor patients differs in the presence or absence of bacterial vaginosis. DESIGN: Observational study. SUBJECTS: One hundred and twelve consecutive patients with objectively confirmed preterm labor admitted to a tertiary care centre were included in the study. Patients with placenta previa, active uterine bleeding or indication for an immediate delivery (e.g. severe pre-eclampsia or suspected fetal asphyxia), or severe fetal anomalies were excluded. METHODS: Transvaginal ultrasonography was used to measure cervical length and internal os width. Bacterial vaginosis was diagnosed by Gram stain of a vaginal smear. RESULTS: A total of 36 patients (32%) had bacterial vaginosis. Cervical length in this group was shorter than in patients with normal flora (mean 20.4 +/- 7.2 mm vs. 26.4 +/- 6.7 mm; P = 0.0002), and more patients with bacterial vaginosis had a dilated internal cervical os > or = 5 mm (67% vs. 30%, P = 0.001). There were no significant differences, however, in preterm delivery rate and birth weight between the two groups; the overall preterm delivery rate was 40%. A cervical length < 25 mm was predictive of preterm delivery (P = 0.001, RR 4.2, 95% CI 1.8-9.7). CONCLUSIONS: These data suggest that cervical change in preterm labor is more pronounced in patients with bacterial vaginosis but without a concomitant increase in the risk for preterm delivery. Despite this association, the cervical length measured by transvaginal ultrasonography alone is a useful predictor of preterm delivery, independent of the presence or absence of bacterial vaginosis. PMID- 10846782 TI - Preoperative diagnosis of the primary fallopian tube carcinoma by three dimensional static and power Doppler sonography. AB - OBJECTIVE: To investigate whether three-dimensional static and power Doppler ultrasound improves the diagnosis of primary Fallopian tube carcinoma. METHODS: During a 2-year period five cases of primary Fallopian tube carcinoma were selected from a cohort of 520 patients with a previous scan suggestive of an adnexal tumor. RESULTS: Tubal malignancy occurred in patients between 49 and 64 years, with presenting symptoms such as pain, vaginal bleeding and leukorrhea. CA 125 was elevated in three cases of tubal carcinoma with stages II and III, while in two patients with stage I, CA 125 was within the normal limits. Two dimensional ultrasound demonstrated sausage shaped cystic masses with papillary projections in two patients and a complex adnexal mass in one patient. Three dimensional ultrasound revealed sausage shaped cystic and/or complex masses with papillary projections in all five cases of tubal malignancy. In one patient preoperative 3-D ultrasound correctly predicted bilateral tumors, while 2-D transvaginal sonography found only unilateral changes. Additional 3-D power Doppler examination depicted vascular geometry typical for malignant tumor vessels such as arteriovenous shunts, microaneurysms, tumoral lakes, blind ends and dichotomous branching in each of the cases with Fallopian tube carcinoma. CONCLUSIONS: Three-dimensional ultrasound allows precise depiction of tubal wall irregularities such as papillary protrusions and pseudosepta. Improved understanding of anatomical relationships may aid in distinguishing ovarian from tubal pathology. Multiple sections of the tubal sausage like structures enable determination of local tumor spread and capsule infiltration. Study of the vascular architecture in cases of Fallopian tube malignancy is further enhanced using 3-D power Doppler imaging. PMID- 10846783 TI - Early prenatal sonographic diagnosis of congenital hypophosphatasia. AB - A pregnant woman of 14 weeks' gestation was sonographically examined due to large for-dates uterine size. The ultrasound examination showed poor ossification of all bony structures. All limbs were shortened with no evidence of fractures. The echodensity approximated that of the surrounding organs. No acoustic shadowing was observed. Based on these sonographic findings, skeletal dysplasia and short limb dwarfism were diagnosed, the most likely condition being congenital hypophosphatasia. Early cordocentesis was successfully performed at 15 weeks' gestation to determine fetal alkaline phosphatase concentration. This was undetectable. The prenatal diagnosis of congenital hypophosphatasia was made. After counselling, the woman decided to opt for termination of pregnancy which was performed vaginally. Post-abortion findings confirmed the prenatal diagnosis. To our knowledge, this is the earliest sonographic diagnosis of this condition reported. PMID- 10846784 TI - Prenatal diagnosis of thrombocytopenia-absent-radius (TAR) syndrome. AB - The prenatal diagnosis of thrombocytopenia-absent-radius (TAR) syndrome using ultrasound and cordocentesis in the 16th week of gestation is established. The sonographic findings detected in this case included bilateral absence of the radius and club hands with normal thumbs and metacarpals. Because of a high index of suspicion for the syndrome, cordocentesis for fetal blood analysis was performed. Thrombocytopenia, with a platelet count of 14,000/mm3, was identified. The pregnancy was electively terminated and subsequent findings confirmed the sonographic diagnosis. This report, to our knowledge, is one of a very limited number of cases published in the literature, in which the prenatal diagnosis of TAR syndrome was made. PMID- 10846785 TI - Prenatal diagnosis of total arhinia. AB - Complete or total arhinia, in which there is absence of the soft tissue of the nose, is extremely rare. The embryological origin of the defect is thought to be maldevelopment of the paired nasal placodes. Available neonatal case descriptions have reported the frequent coexistence of other facial anomalies. This report is the first to describe the antenatal diagnosis of total arhinia confirmed after delivery. Sonographic views of the fetal profile, showing an absence of the nose led to consideration of the diagnosis. PMID- 10846786 TI - In search for the 'third point'. PMID- 10846787 TI - Prenatal diagnosis of fetal microcephaly: contribution of power Doppler. PMID- 10846788 TI - Results of expectant management of women early pregnancy and unknown location. PMID- 10846789 TI - Re: The clinical significance of fetal isolated cerebral borderline ventriculomegaly: report of 31 cases and review of the literature. Ultrasound Obstet Gynecol 1999; 14(5):320-6. PMID- 10846790 TI - Picture of the month. Antenatal diagnosis of placenta percreta using gray-scale ultrasonography, color and pulsed Doppler imaging. PMID- 10846791 TI - Application of derivative UV spectrophotometry for the determination of ciprofloxacin, norfloxacin and ofloxacin in tablets. AB - The first-, second-, third- and fourth-order derivative spectrophotometric methods, by using the "peak-zero" (P-0) and "peak-peak" (P-P) techniques of measurement have been developed for the determination of ciprofloxacin hydrochloride, norfloxacin and ofloxacin in tablets. The calibration curves are linear within the concentration range of 2.0-12.0 micrograms ml-1 for ciprofloxacin hydrochloride, 1.0-10.0 micrograms ml-1 for norfloxacin and 2.5 15.0 micrograms ml-1 for ofloxacin. The procedure is simple, rapid and the results are reliable. PMID- 10846792 TI - TLC fractionation and visualization of selected phenolic compounds applied as drugs. AB - Sixteen visualizing agents, thirteen of which were a group of alkacemetric indicators were used for the detection of selected drugs (being phenolic compounds): adrenaline, dopamine, phenylephrine, metaraminol, fenoterol, and bithionol. Visualizing effects for the drugs investigated after their TLC separation were estimated on silicagel (A), mixture silica gel/kieselguhr (B), and on polyamide (C). The best separations and the most positive visualising effects were obtained on adsorbent A, and the least on absorbent C. On adsorbent A, the most profitable detectability equal to 100 ng, was obtained also for adrenaline, dopamine and fenoterol with the application of basic solution of bromocresol green and brilliant cresyl blue as visualizing agents. PMID- 10846793 TI - Densitometric determination of impurities in pharmaceuticals. Part VI. Determination of 4,4-bis[4-(p-chlorophenyl)- 4-hydroxypiperidino]butyrophenone in haloperidol. AB - A chromatographic and densitometic method for identification and quantitative determination of 4,4-bis[4-(p-chlorophenyl)-4-hydroxypiperidino]butyrophenone as an impurity in haloperidol pharmaceutical has been developed. The HPTLC plates and chloroform-methanol-ammonium hydroxide 25% (90:9:1) were used for chromatographic separation as stationary and mobile phases respectively. Detection has been carried out in UV at lambda = 350 nm. The determination could be made directly without preliminary component separation by extraction. Based on the statistical analysis of obtained results, it was found that the new method is accurate and repeatable. PMID- 10846794 TI - Redox methods validation of paracetamol and ascorbic acid in pharmaceutical preparations. AB - The contents of active substances were determined in a preparation TP-4 (tablets) containing paracetamol, ascorbic acid, caffeine and phenylephrine hydrochloride. For the determination of paracetamol and ascorbic acid a non-specific (cerometric and titration of 2,6-dichloroindophenol) method based on a redox reaction was used. Validation of the methods, performed on model mixtures, proved those methods to be accurate, precise, reproducible and linear within the range from 50% to 150% of the amount declared in the preparation. The content of paracetamol and ascorbic acid in TP-4, Thompayrin, Panadol Extra, Ring N, Polopiryna C, Efferalgan Vitamin C and Vitaminum C 0.2 satisfies the FP V demands (+/- 10% of the declared amount). PMID- 10846795 TI - Comparative dissolution testing of paracetamol commercial tablet dosage forms. AB - Dissolution can best be described as a tool that can provide valuable information about the availability of a drug product. In this study, nine different paracetamol tablet dosage forms available on the Turkish Drug Market have been investigated and physical controls were realized. Paddle and rotating basket apparatus methods were applied to all the formulations. In order to evaluate the dissolution rates, five different kinetics have been studied and the best fitting kinetics was found to be the Hixson-Crowell kinetics. It was found that all the preparations are in accordance with the Pharmacopeia standards. PMID- 10846796 TI - Physico-chemical properties and hydrolysis of oxprenolol octanoyl and benzoyl esters. AB - The kinetics of hydrolysis of octanoyl ester of oxprenolol (O-OXP) and benzoyl ester of oxprenolol (Benz-OXP) has been investigated in aqueous solution at 310 K over the pH range 0.42-9.5. The decomposition was followed by UV spectral method. At the pH range 0.42 to 9.5, hydrolysis of oxprenolol esters (E-OXP) consists of hydrolysis of BH+ molecules catalyzed by hydrogen ions, spontaneous hydrolysis of BH+ molecules and hydrolysis of BH+ and B molecules catalyzed by hydroxide ions. Various buffer substances were found to exhibit general acid and base catalysis of the degradation. PMID- 10846797 TI - Effects of iron and copper ions on the inducibility of hydrogen peroxide in E. coli. AB - Inducing capacities of hydrogen peroxide in SOS chromotest were investigated in the absence and in the presence of iron and copper ions. Strain PQ37 of Escherichia coli and two strains with altered protection against oxidative damage (PQ300 and OG400) were employed. The latter two strains were more sensitive to the hydrogen peroxide inducing activity than strain PQ37. At the concentration of 150 nmol/ml and 300 nmol/ml, iron ions induced SOS response. At the concentration range from 30 nmol/ml to 300 nmol/ml iron enhanced the effects produced by H2O2. Results with copper were negative. PMID- 10846798 TI - Synthesis and study of the antischistosomal potency and induced biological parameters of a new 2-palmitoyl analogue of the universal antihelminthic praziquantel. AB - 2-Palmitoyl[1,2,3,6,7,11b]hexahydro-4H-pyrazino[2-la]isoquinoline-4-one [III] a highly lipophilic analogue of the universal antihelminthic PRAZIQUANTEL [I] was rationally multi-stepwise synthesized and antischistosomally and biochemically screened. The 2-palmitoyl conjugation was hypothesized to be an antischistosomal adjuvant (Tween 40 mimicry), to the reported crucial pyrazino-isoquinoline moiety. On a constant weight doses bases of I and III (500 mg/kg mouse body weight), the activity of III was found to be approximately 70% of I (mice infected with S. mansoni cercariae) and with satisfactory toxicological and biochemical profile (mice liver and kidney functions). Equivalent molar weight assay (M 1:1.4 for I and III, respectively), which could further plead in favour of the potency of III, was not yet tested. The analogue III, which favourable incorporates the human metabolically and physiologically compatible, palmitic acid segment, seems to be an antischistosomally promising candidate for more integrated studies. PMID- 10846799 TI - Investigation of diazepam lipospheres based on Witepsol and lecithin intended for oral or rectal delivery. AB - Diazepam was incorporated in lipospheres prepared by high pressure homogenization of melted Witepsol (10%) dispersed in aqueous lecithin (2.4%). Diazepam content was 0.4% and more than 98% of the dose was found to be encapsulated in the lipospheres. Although the initial mean particle size was 0.3 micron, the liposhperes agglomerated during storage and this phenomenon was not eliminated by increasing lecithin concentration to 4% or incorporation of oleic acid (0.1%) and co-surfactants, polysorbate 80 (0.5%) or poloxamer (up to 6%). The formulation was not able to mask effectively bitter taste of diazepam, even when lipids of higher melting temperature, namely glyceryl tripalmitate or stearic acid, were introduced. PMID- 10846801 TI - Qualitative and quantitative analysis of phenolic acids in Asclepias syriaca L. AB - TLC and HPLC was applied to qualitative and quantitative determination of phenolic acids free and liberated by acid and alkaline hydrolysis in common milkweed. The presence of phenolic acids, namely p-hydroxybenzoic, p-coumaric, protocatechuic and caffeic acids, was confirmed in the leaves and flowers. Moreover, the flowers contained gallic acid, and the leaves contained alpha resorcylic, vanilic and chlorogenic acids. Conjugated forms of phenolic acids predominate in the plant. They hydrolyse mainly to coffeic, p-coumaric, ferulic and p-hydroxybenzoic acids. PMID- 10846800 TI - Iridoids and phenylethanoids of Verbena bipinnatifida nutt. AB - Iridoids and phenylethanoids were separated from Verbena bipinnatifida Nutt. (Verbenaceae). The compounds identified are pulchelloside I and II--iridoids and martynoside, verbascoside-phenylethanoids. The structures of all the compounds isolated were elucidated by classical and spectroscopic methods. PMID- 10846802 TI - Influence of alpha-escin on skeletal changes in ovariectomized rats. AB - The aim of the present study is to investigate the influence of alpha-escin (35 mg/kg per os daily; administered for 4 weeks) on the development of osteopenia caused by bilateral ovariectomy in 3-month-old female Wistar rats. The experiments were carried out on four groups of animals: I (C)--control sham operated rats, II (OVX)--ovariectomized rats, III (E)--sham operated rats which were administered alpha-escin, IV (OVX + E)--ovariectomized rats which were given alpha-escin. In all groups body weight growth, masses, length and tibia diameter were determined as well as histological specimens of right femur and tibia slices were used for histometric measurements including: the transverse cross-section area of the tibial shaft cortex, transverse cross-section area of the tibial marrow cavity, periosteal osteoid width, endosteal osteoid width, periosteal thickness growth in the tibia, endosteal thickness growth in the tibia, epiphyseal and metaphyseal trabeculae thickness in the femur and epiphyseal cartilage width. Bilateral ovariectomy in matured female rats caused osteopenic skeletal changes. alpha-Escin (35 mg/kg per os daily) administered to the ovariectomized rats for the following 28 days decreased only a little the development of osteopenic skeletal changes caused by bilateral ovariectomy. PMID- 10846803 TI - Methylation of 4-hydroxycoumarin with diazomethane. AB - 4-Hydroxycoumarin was methylated with diazomethane. A mixture of 4 methoxycoumarin and 2-methoxychromone was separated chromatographically. PMID- 10846804 TI - Dysfunctional meaning of posttraumatic intrusions in chronic PTSD. AB - The paper suggests that the negative idiosyncratic meaning of posttraumatic intrusions (e.g., 'I am going crazy') and cognitive strategies intended to control the intrusions play a major role in maintaining posttraumatic stress disorder. Two studies of 159 and 138 motor vehicle accidents survivors showed that the dysfunctional meaning of intrusions explained a proportion of the variance of the intrusion-related distress, strategies used to end the intrusions, and PTSD severity that was not explained by intrusion frequency, accident severity, or by general catastrophic thoughts when anxious. Rumination, thought suppression, and distraction when having intrusions showed substantial correlations with PTSD severity, as did avoidance of reminders of the accident. The results have implications for the treatment of chronic PTSD. PMID- 10846805 TI - Evoking analogue subtypes of panic attacks in a nonclinical population using carbon dioxide-enriched air. AB - The increasing recognition that panic attacks are heterogeneous phenomena necessitates better and more objective criteria to define and examine what constitutes a panic attack. The central aim of the present study was to classify subtypes of panic attacks (i.e. prototypic, cognitive, and non-fearful) in a nonclinical sample (N = 96) based on the concordance/discordance between subjective and physiological responding to multiple inhalations of 20 and 13% CO2 enriched air. Results show that a substantial proportion of this nonclinical sample (55.2%) responded to the CO2 challenge in a manner consistent with clinical and research definitions of different subtypes of panic attacks. The implications of this dimensional approach for discriminating subtypes of panic in the laboratory are discussed as a means to better understand the phenomenology and nature of panic attacks. PMID- 10846806 TI - Changes in the magnitude of the eyeblink startle response during habituation of sexual arousal. AB - Modulation of the startle response was used to examine emotional processing of sexual stimulation across trials within a session. Eyeblink startle was elicited by a probe (burst of intense white noise) presented intermittently while men were viewing an erotic film segment. Repeated display of the film segment resulted in a progressive decrease in sexual arousal. Habituation of sexual arousal was accompanied by a reduction over trials in the extent the men felt absorbed when viewing the erotic stimulus and by an increase over trials in the magnitude of the eyeblink startle response. Replacing the familiar stimulus by a novel erotic stimulus increased in sexual arousal and absorption and reduced startle (novelty effect), while dishabituation was evident for all three response measures when the familiar stimulus was reintroduced. This pattern of results indicates that with repeated presentation an erotic stimulus is experienced not only as less sexually arousing but also as less appetitive and absorbing. The question of whether habituation of sexual arousal is mediated by changes in attentional and affective processing over trials is discussed, as are clinical contexts in which eyeblink startle can be used in studying aspects of sexual functioning. PMID- 10846807 TI - Danger expectancies, self-efficacy and insight in spider phobia. AB - In the current edition of the Diagnostic and Statistical Manual of Mental Disorders (DSM-IV; American Psychiatric Association, (1994) Diagnostic and statistical manual of mental disorders (4th ed.) Washington, DC: author) phobic adults and adolescents are said to "recognize that the phobia is excessive or unreasonable" given the actual danger posed by the feared situation. The present study examined perceptions of danger in 15 spider phobic subjects and a matched set of controls before, during and after a spider-avoidance test. When detached from the phobic stimulus, phobic subjects: (1) gave higher estimates of the probability of being bitten than controls did; (2) gave higher estimates of the injuries that would result from being bitten and (3) in line with these first two findings, believed their high levels of anticipated anxiety were more reasonable and appropriate to the demands of the situation than controls did. These findings are inconsistent with the prevailing notion that when detached from the phobic situation patients can accurately evaluate the danger of potential phobic encounters. Instead, the findings suggest that phobic individuals, whether detached or in the presence of the feared object, have relatively limited insight into the irrationality of their fears. In examining the mediation of phobic phenomena, both self-efficacy and danger estimates remained significantly related to the anxiety and avoidance experienced in the spider-avoidance task. Further research designed to experimentally establish the likely causal roles of these two constructs is warranted. PMID- 10846808 TI - Recurrent images and early memories in social phobia. AB - A recent model [Clark, D. M. & Wells, A. (1995). A cognitive model of social phobia. In R. Heimberg, M. Liebowitz, D. A. Hope & F. R. Schneier (Eds.), Social phobia: diagnosis, assessment and treatment (pp. 69-93). New York: Guildford Press] suggests that a distorted image of one's public self lies at the heart of social phobia. A previous study of spontaneous imagery [Hackmann, A., Surawy, C. & Clark, D. M. (1998) Seeing yourself through others' eyes: a study of spontaneously occurring images in social phobia. Behavioural and Cognitive Psychotherapy, 26, 3-12] confirmed that patients with social phobia frequently report experiencing negative, distorted, observer-perspective images when in anxiety provoking social situations. In the present study, 22 patients with social phobia were given a semistructured interview which aimed to further explore the nature of social phobic imagery. All participants were able to identify negative spontaneous images that were recurrent in the sense that their content appeared to be relatively stable over time and across different feared social situations. Most recurrent images involved several sensory modalities. Most recurrent images were linked to memories of adverse social events that clustered in time around the onset of the disorder. Taken together, the results suggest that in patients with social phobia, early unpleasant experiences may lead to the development of excessively negative images of their social selves that are repeatedly activated in subsequent social situations and fail to update in the light of subsequent, more favourable experiences. Implications of the findings for the understanding and treatment of social phobia are discussed. PMID- 10846809 TI - Post-event processing in social anxiety. AB - A psychometric study was conducted in order to collect basic information about post-event processing in social anxiety. It was found that such processing occurs commonly after an anxiety-evoking or embarrassing social event and post-event processing scores were significantly correlated (r = 0.40) with social anxiety. The recollections of the social event tended to be recurrent and intrusive, interfering with concentration. Post-event processing was associated with the avoidance of similar social situations. The results are discussed in terms of the Clark and Wells model of social phobia. PMID- 10846810 TI - Gender differences in susceptibility to posttraumatic stress disorder. AB - We examined data from a community survey of trauma exposure and DSM-IV PTSD in Winnipeg, Canada to explore factors associated with the higher rate of PTSD in women than men. Women were found to be at significantly increased risk for PTSD following exposure to serious trauma (odds ratio approximately 5), even when sexual trauma--which predominates in women--was excluded (odds ratio approximately 3). Adjusting for gender differences in the number of lifetime traumata, or in the likelihood of the trauma being associated with particular reactions to or consequences of the event (i.e. thinking that one would be killed or seriously injured; sustaining a serious physical injury; seeing someone else seriously injured or killed) did not result in a lessening of the PTSD risk in women. Women were found to be at increased risk for PTSD following nonsexual assaultive violence (e.g. mugging or other physical attack) but not following non assaultive trauma (e.g. fire, witnessing injury to others). Understanding the basis of (and parameters for) this increased susceptibility to PTSD in women compared to men following trauma exposure should be a priority for future traumatic stress research. PMID- 10846811 TI - [Porcine stress syndrome and PSE meat: clinical symptoms, pathogenesis, etiology and animal rights aspects]. AB - A review is given about the clinical symptoms, pathogenesis and aetiology of the porcine stress syndrome, furthermore aspects of animal welfare are discussed. The current breeding programmes of pig industry in Germany in many cases include animals with a mutation of the ryanodine-receptor (RYR-1)-gene--homozygous or heterozygous. This situation is the result of an intensive breeding of pigs during the last decades with the intention of increased lean carcass content and corresponding proceeds. The homozygous pigs are more stress susceptible (porcine stress syndrome) and produce meat of poor quality (PSE), which is also the case to some extend in heterozygous animals. The clinical symptoms of this muscle disease are characterised by a deficit of oxygen and a rapid glycolysis accompanied by a production of lactic acid and acidosis primarily in II B white muscle fibres. There is no doubt that a very close causal relation exists between the mutation of the RYR-1 and the porcine stress syndrome as well as the poor meat quality. The present knowledge of this disease, the genetic background, the physiology and pathophysiology of the mutation of the RYR-1 leads to the imperative conclusion to eliminate this mutated RYR-1 by selection of healthy pigs, which has been done successfully in other countries with important pig production. This conclusion is also supported by simple economic reasons because fertility, reproduction and daily weight gain are significantly reduced in stress susceptible pigs. Furthermore, it should be emphasised that regular breeding with the mutated RYR-1 is also a matter of animal welfare. The evident correlation between the mutated RYR-1 and the porcine stress syndrome, which includes degeneration of the muscle, pain and even life threatening malignant hyperthermia, can easily lead to the accusation in the public that diseased animals are used for pig meat production. Consequently, the authors would like to urge the breeding companies and the responsible authorities to discuss the problem with the intention to finish the current breeding programmes using animals with the mutated RYR-1 within a reasonable period of time. PMID- 10846812 TI - [Results of the German investigation on the EU project "Salmonella in Pork (Salinpork)"--1. Investigations in the farms]. AB - Reports of human salmonellosis caused by the consumption of pork and the introduction of control and surveillance programmes in different countries of the European Union were the reason for carrying out an international study under the title "Salmonella in Pork (Salinpork)": Six different EU-countries were involved in this study, which was supported by the Commission of the European Community and which was carried out over a period of April 1996 to April 1999. The aim of the investigation presented was to determine the prevalence of Salmonella in fattening, breeding and farrow-to-feeder herds as well as the determination of risk factors for the introduction of Salmonella into the farm (Part 1). In addition, sources of contamination of pork should be detected by taking samples of the product and the environment in the slaughterhouse (Part 2). In Germany, the investigation into Salmonella infections of 60 fattening, 20 breeding and 20 farrow-to-feeder herds were carried out in Schleswig-Holstein. The investigation included bacteriological examinations of feed and faecal samples for sero- and phagetyping and serological examinations by using the Danish Mix-ELISA. From 2,947 serological investigated fattening pigs were 7.3% (n = 213) positive, from 797 breeding sows 9.2% (n = 73) were serological positive and 4.5% (n = 18) of the investigated sows (N = 399) in farrow-to-feeder herds were serological positive. Altogether, 28.3% of the fattening, 50.0% of the breeding and 15.0% of the farrow-to-feeder herds were serological positive. A questionnaire was used to capture data about management, hygiene measures, feeding systems and the occurrence of diseases in the herd. After statistical analysis the common risk factor of fattening herds and sow herds was the use of pelleted feed. But in a control study with 17 different fattening herds the result could not be proven. Other factors which can influence the occurrence of Salmonella infections were discussed. PMID- 10846813 TI - [Epizootiology of fowls adenoviruses]. AB - Fowl adenoviruses of the serotype 4 from Germany were characterised by restriction enzyme analysis in comparison to isolates from Asia, South America and the FAV4 reference strain KR5. Only strain Da60 which was isolated from a psittacine aviary was identical with the reference strain KR5. None of the isolates was identical with the highly pathogenic strains from India and Ecuador. One-day-old chicks were infected orally and intramuscularly with the reference strain KR5, the psittacine isolate Da60 and isolate K1013 from Ecuador. Whereas no mortality was seen with the two strains KR5 and Da60, the mortality with K1013 was 100%. The main pathological signs were a swollen liver with necrosis and a lymphocyte depletion with a loss of the follicle structure. To investigate a second subject of avian adenovirus epizootiology several FAVs were characterized serologically and with PCR which was combined with the digestion of the PCR products. Including the reference strains, both methods were compared. It was shown that the digestion of the PCR products allows a clear attribution to a specific serotype, which underlines the usefulness of this method for diagnostic purposes. PMID- 10846814 TI - [Is there a predictable correlation between time of birth, birth and the puerperal period and clinical and hormonal findings in horses?]. AB - The investigations were done on 108 mares (78 warm-blooded horses, 25 cold blooded horses and 5 trotters). To pursue the question in the title, progesterone and estradiol values in jugular vein-blood were examined starting from 30 days before the calculated date of parturition until 30 days after that event. In parallel to that Ca/Mg ratio was measured by means of a semiquantitative test strip in the milk. Clinical signs of the birth, parturition itself and the post partum period were recorded. In respect to the P4 and E2 values it was that the P4:E2 ratio changed towards parturition in favour to E2 which was interpreted as a sign for the beginning of labour contractions as well as a signal to the ovarian hormonal activity and feed-back regulation. The "predict of the foal test" by measuring Ca/Mg ratio is helpful only in combination with clinical findings. This test can be used for the decision to induce parturition. Concerning the colour and viscosity of the milk, it was observed to change from grey-aqueous 48 h a.p. through grey-brownish 24 h a.p. to white-yellow and viscous 12 h a.p. The results show that heavy mares more often retain the fetal membranes. They often bear lighter offspring with equally lighter fetal membranes, and the surfaces of the microvilli of the microcotyledonas seem to be smaller. Beside this it is concluded that during pregnancy disturbances in the fetomaternal relations probably exist as a result of a deficiency in placental development that finally results in retention secundinarum. PMID- 10846815 TI - Parvovirus B19 infection and its significance in pregnancy. AB - Parvovirus B19 causes prolonged epidemics of erythema infectiosum, particularly in primary school-aged children. Infection causes clinically significant anaemia in individuals with high red cell turnover, including the fetus. Approximately 40% of women of childbearing age are susceptible, and annual seroconversion rates vary from 1.5% during endemic periods to 10-15% during epidemics. Infection occurs in around 50% of susceptible women exposed at home and 20-30% following occupational exposure (for example, at a primary school). Maternal infection in the first half of pregnancy is associated with 10% excess fetal loss and hydrops fetalis in 3% of cases (of which up to 60% resolve spontaneously or with appropriate management). No congenital abnormalities or long-term sequelae have been attributed to parvovirus B19 infection. The overall risk of serious adverse outcome from occupational exposure to parvovirus B19 infection during pregnancy is low (excess early fetal loss in 2-6/1,000 pregnancies and fetal death from hydrops in 2-5/10,000 pregnancies). It is not recommended that susceptible pregnant women be excluded routinely from working with children during epidemics. PMID- 10846816 TI - Disease activity in Victoria. PMID- 10846817 TI - Detection of the exotic mosquito Culex gelidus in the Northern Territory. PMID- 10846818 TI - Communicable diseases surveillance. PMID- 10846819 TI - [Study of the inclination angle between the edentulous upper and lower alveolar ridge in the sagittal plane]. AB - Angle of inclination between the upper and lower edentulous alveolar ridge in the sagittal plane was examined on 72 patients (55 woman and 17 men) in centric occlusion. Photos were taken by a special camera (Polaroid MACRO 5 SLR) of the functional casts mounted in average value articulators, on a special squared film, from the left and right side. The angle between the lines connecting mesially the midline of the alveolar ridge, distally the highest point of the tuber alveolar maxillae and the line drown through the stop-lines, respectively was measured on both sides with 1 degree precision. Distally open angle (max. 7 degrees) was found in 11% of the examine group. The alveolar ridge lines were parallel in only 3% of the cases. Mesially open angle (max. 20 degrees) was found at 86%. Statistical analysis has shown, that there is a significant difference between men and women as well as the left and right side. PMID- 10846820 TI - [Use of the TRI AUTO ZX instrument in clinical endodontics]. AB - The aim of this study was to test the clinical reliability and applicability of TRI AUTO ZX (TAZX) instrument for working length determination and shaping as well as widening of root canals. Thirty teeth were treated by the means of TAZX instruments. A close correlation was found between working length data obtained by TAZX instrument and by conventional radiography. Using TAZX no failure (file separation, apical foramen transposition, etc.) was observed. According to our experiences TRAZX instrument seems to be a valuable tool for endodontic procedures. PMID- 10846821 TI - [Surgical repair of pharyngocutaneous defects resulting from actinomycosis]. AB - The pharyngo-cutan defect is a frequent complication of oropharynx and larynx operations. The not frequent serious actinomycotic inflammation of the neck, and in its treatment the repeated surgical incision was in the background of the defect. The authors review the possibilities of closing these defects, and they describe the method used. PMID- 10846822 TI - [Biomechanical study on bone grafts harvested from cadavers]. AB - Today the technique of free revascularized bone grafts in reconstruction surgery of the head and neck has been well developed. The grafts can be harvested from different parts of the body. No data have been found regarding biomechanics of different grafts. The authors examined 18 samples of cadaver iliac crest, radius, lateral edge and spine of scapula and fibula, respectively, and 6 samples of mandible. The study pointed out that the stiffness of the fibula approached those of mandible. The stiffness of the other bone grafts were found much smaller. PMID- 10846823 TI - mi-transcription factor as a regulator of mast cell differentiation. AB - Masts cells are progeny of the hematopoietic stem cell. For the differentiation of mast cells, a transcription factor encoded by the mouse mi locus (MITF) plays an important role. The expression of many genes encoding proteins that are essential for the function of mast cells is regulated by MITF. Because various mutant mice are available at the mi locus and because cultured mast cells are easily obtained from the spleen of these mutant mice, this system may be a good model for studying the regulation of hematopoietic cell differentiation by a transcription factor. PMID- 10846824 TI - Ex vivo expansion of megakaryocytic cells. AB - The use of platelet transfusion to ensure the recovery of thrombopoiesis in patients constitutes high-cost support. The identification and cloning of recombinant human thrombopoietin (TPO) and the development of efficient methods of purification of hematopoietic stem cells and progenitor cells have ameliorated the development of strategies of ex vivo expansion of megakaryocyte (MK) progenitor cells and mature MKs. Synergistic combinations of cytokines including TPO, interleukin (IL)-1, IL-3, IL-11, stem cell factor, and FLT-3 ligand induce the ex vivo expansion of colony-forming unit-MK progenitors and MKs from cytokine mobilized peripheral blood cells, bone marrow, and cord blood CD34+ cells. Depending on the various culture conditions, i.e., combinations of growth factors, initial concentration of CD34+, serum or serum-free cultures, and/or oxygen tensions, the expansion-fold of MKs and their progenitor cells vary greatly. The clinical applications of the reinfusion of ex vivo-generated MK cells have been investigated successfully in cancer patients following high-dose chemotherapy. This review reports the latest information concerning ex vivo expansion of MKs and the current status of clinical trials. PMID- 10846825 TI - Prognostic and predictive significance of p53 mutation in aggressive B-cell lymphoma. AB - p53 protein plays a central role in cellular responses, including cell-cycle arrest and cell death in response to DNA damage such as that caused by many chemotherapeutic agents and radiation used in cancer therapy. p53 dysfunction can induce abnormal cell growth, increased cell survival, genetic instability, and drug resistance. p53 mutations occur in approximately half of human cancers. They have been found in about 20% of patients with aggressive B-cell lymphoma and are associated with poor treatment outcome and poor prognosis. The purpose of this review is to provide an outline of the biological functions of p53 and assess the role of p53 in clinical settings. PMID- 10846826 TI - Hemoglobin Pitie-Salpetriere [beta 34 (B16) Val-->Phe] showing erythrocytosis and mild hemolysis in a Japanese man. AB - We report the first case of Hemoglobin Pitie-Salpetriere (Hb P-S) identified among the Japanese population. The patient was a 33-year-old man referred to us because of severe erythrocytosis and mild hemolysis. DEAE high-performance liquid chromatography showed an abnormal broad peak around Hb A2 peak. Isoelectrofocusing detected abnormal Hb at the position of the Hb F band, and the content of abnormal Hb was estimated at about 25%. An instability test according to the isopropanol precipitation method was positive, and the beta/alpha ratio of biosynthesized globin was slightly reduced. Structural analyses demonstrated the substitution of phenylalanine for valine at beta 34, which was also confirmed by DNA sequencing; that is a single base substitution of GTC-->TTC at codon 34 of beta chain. From these findings, the abnormal Hb was identified as being a high oxygen-affinity variant, Hb P-S (beta 34 [B16] Val-->Phe). Hb P-S was detected in the patient's mother but not in his father, suggesting that the inheritance pattern is autosomal dominant. It was suggested that the slightly unstable state of Hb P-S caused by the looseness of alpha 1 beta 1 contact could result in mild hemolysis. PMID- 10846827 TI - A novel nonphagocytic mechanism of erythrocyte destruction involving direct cell mediated cytotoxicity. AB - The number of erythrocytes fell when co-cultured with cell preparations derived from mouse spleen, thymus, bone marrow, or peritoneal exudate (PE) cells. Erythrocyte-depletion activities (EDA) of different leukocyte preparations were in the order PE > spleen > thymus > bone marrow. Adherent, nonadherent, T depleted, and T-enriched cell subpopulations had comparable EDA. Spleen cells from athymic nude mice, however, lacked significant EDA. In addition, EDA was boosted by Concanavalin A (Con A) but not by lipopolysaccharide, indicating that T cells may play a crucial role in inducing EDA in spleen cells. Paraformaldehyde fixed spleen or PE cells, as well as membrane preparations isolated from spleen cells, efficiently lysed erythrocytes. Erythrocyte ghost membranes inhibited erythrocyte lysis by control or paraformaldehyde-fixed spleen cells. Treatment with hamster anti-mouse Fas or anti-mouse tumor necrosis factor receptor (TNFR) antibody could opsonize erythrocytes for faster depletion by spleen cells, suggesting an expression of Fas and TNFR on erythrocytes. TNF alpha could lyse erythrocytes in a dose-dependent fashion. Additionally, enhanced spleen cell EDA induced in response to succenyl Con-A could be blocked by anti-TNF alpha antibodies. Our results provide evidence for a direct cell-mediated cytotoxicity (CMC) of erythrocytes by leukocytes. A role of molecules of Fas and the TNF family in CMC of erythrocytes has also been suggested. Further work is needed to understand if, and to what extent, CMC of erythrocytes contributes to erythrocyte destruction in vivo and to determine its patho-physiological significance. PMID- 10846828 TI - Low-dose cytarabine and aclarubicin in combination with granulocyte colony stimulating factor (CAG regimen) for previously treated patients with relapsed or primary resistant acute myelogenous leukemia (AML) and previously untreated elderly patients with AML, secondary AML, and refractory anemia with excess blasts in transformation. AB - We used the CAG regimen (low-dose cytarabine [10 mg/m2 per 12 hours, days 1-14], aclarubicin [14 mg/m2 per day, days 1-4], and granulocyte colony-stimulating factor [200 micrograms/m2 per day, days 1-14]) for the treatment of patients with primary resistant acute myelogenous leukemia (AML) and previously untreated elderly patients with AML, secondary AML, and refractory anemia with excess blasts in transformation (RAEB-T) in addition to relapsed AML. Forty-three of 69 (62%) patients achieved complete remission (CR), including 29 of 35 (83%) patients with relapsed AML, 1 of 8 patients with primary resistant AML, 5 of 8 elderly patients with previously untreated AML, and 8 of 18 patients with previously untreated secondary AML or RAEB-T. Ten of 22 (45%) patients > or = 65 years old achieved CR. The patients who achieved CR received at least 1 course of modified CAG therapy as the first consolidation therapy, followed by various second consolidation and intensification therapies. The median disease-free survival and overall survival were 8 and 15 months, respectively, for relapsed AML; 11 and 8 months for the elderly patients; and 8 and 17 months for secondary AML and RAEB-T. Myelosuppression was mild to moderate, and other than fever, severe nonhematologic toxicity was rare. CAG as the induction therapy seems promising for the treatment of various categories of poor-prognosis AML. PMID- 10846830 TI - Low proportion of G0-phase cells during induction chemotherapy correlates with subsequent remission in acute myeloid leukemia. AB - Kinetic resistance is assumed to be one of the main mechanisms of drug resistance in acute myeloid leukemia (AML), but the relationship between cell cycle status at diagnosis and achievement of complete remission (CR) is controversial. Based on the possibility that the cell cycle data after starting induction chemotherapy are more important than the pretreatment data, we used 3-color flow cytometry to examine the cell cycle (G0, G1, S, and G2/M phases) of AML cells on days 0, 5, and 9 of the first induction chemotherapy in 20 patients. Cell cycle data at these 3 time points were compared in the patients who achieved CR (CR cases) and in the patients who had persistent leukemia (non-CR cases) after the induction chemotherapy. In the CR cases, there was a tendency for the percentages of G0 phase AML cells on days 5 and 9 to be smaller than that on day 0, while the opposite tendency was observed in the non-CR cases. When cell cycle data were compared between the CR and non-CR cases, the percentage of G0-phase AML cells on day 9 differed significantly (CR cases 6.9% +/- 10.9%, non-CR cases 50.1% +/- 38.4%; P = .0024). This significance remained when the patients' AML subtype was taken into consideration. None of the other cell cycle data at each time point or the hematologic parameters, which may be related to CR achievement, showed differences between the CR and non-CR cases. We emphasize the importance of cell cycle analysis after initiating therapy and suggest that such analysis can identify refractory AML subjects. The identified subjects may be candidates for clinical trials of cell cycle modulators. PMID- 10846829 TI - MLL gene rearrangement in t(9;11) acute myelogenous leukemia with minimal myeloid differentiation (FAB subtype M0). AB - A 25-year-old man was diagnosed with acute myelogenous leukemia (AML), French American-British (FAB) subtype M0, based on cytochemical and flow cytometric findings. Cytogenetic analysis revealed the chromosome translocations t(9;11)(p22;q23), and MLL gene rearrangement was identified by Southern blotting. In adult AML, MLL gene rearrangement was initially reported in FAB M4 and M5 cases, and recently in M1 and M2 cases, but was rare in M0 or M3 cases. Because the sensitivity of detecting MLL gene rearrangement by cytogenetic analysis is extremely low compared with Southern blotting analysis, the MLL gene may be involved in substantial numbers of adult AML cases, regardless of FAB subtype. PMID- 10846831 TI - Fatal hepatic failure caused by chemotherapy-induced reactivation of hepatitis B virus in a patient with hematologic malignancy. AB - A patient with hematologic malignancy and hepatitis B virus (HBV) infection received chemotherapy containing a glucocorticoid. The patient developed severe hepatitis after chemotherapy and, despite achieving complete remission of the malignancy, died of hepatic failure. We carried out a retrospective study of changes in the serological markers of HBV in this patient. Both serum hepatitis B surface antigen (HBsAg) and hepatitis B surface antibody (HBsAb) were negative on admission. During the course of chemotherapy, HBsAg gradually became positive, but no liver dysfunction was apparent until after completion of the chemotherapy. The patient showed no initial evidence of being a latent HBV carrier. Therefore, we believe that screening for HBsAg is insufficient for detecting latent HBV carriers, and that investigation for hepatitis B core antibody (HBcAb) is essential. PMID- 10846832 TI - An adult patient with hypersensitivity to mosquito bites developing mantle cell lymphoma. AB - Hypersensitivity to mosquito bites (HMB) has been known to occur exclusively in the first 2 decades of life and is frequently associated with Epstein-Barr virus (EBV) infection and lymphoproliferative diseases. We report here the first adult patient with HMB, a 61-year-old Japanese man who developed mantle cell lymphoma. EBV was detected in the lymph node by polymerase chain reaction and by in situ hybridization. Serum levels of interleukin (IL)-4, IL-6, and IL-10 were markedly increased, and the T-helper cell (Th)1/Th2 balance determined by intracellular cytokine levels was polarized to Th2. These findings suggest that the Th1/Th2 imbalance could partly be involved in the pathogenesis of HMB. PMID- 10846833 TI - Fatal Epstein-Barr virus-associated hemophagocytic lymphohistiocytosis with clonal karyotype abnormality. AB - We report a case of Epstein-Barr virus (EBV)-associated hemophagocytic lymphohistiocytosis (HLH) with clonal karyotype abnormality. A 5-year-old boy was admitted to our hospital with persistent high-grade fever, hepatomegaly, and pancytopenia. Laboratory data disclosed a coagulation abnormality and severe liver damage. Clonal proliferation of EBV-infected cells was detected in the bone marrow by Southern hybridization, and bone marrow cells exhibited clonal chromosomal abnormality. Although the patient was treated with immunochemotherapy according to the HLH94 protocol, the disease recurred during the induction therapy, and the patient died of disseminated intravascular coagulopathy. Considering this aggressive and fatal clinical course, it is important to take intensive therapeutic measures if karyotype abnormality is noted in the treatment of EBV-HLH patients. PMID- 10846834 TI - Surveillance of the serum Candida antigen titer for initiation of antifungal therapy after post-remission chemotherapy in patients with acute leukemia. AB - The early diagnostic efficacy of serial Candida antigen detection by the assay kit CAND-TEC (a latex particle agglutination test) was evaluated in 12 episodes in 10 patients with acute leukemia after post-remission chemotherapy. To determine the timing to initiate antifungal chemotherapy, we performed the CAND TEC assay serially in each patient. When the patients revealed febrile neutropenia after antileukemic chemotherapy and the Candida antigen titer was increased compared to that measured before the antileukemic chemotherapy (even if the increased titer was at a lower level, e.g., from negative to 1:1 positive or from 1:1 to 1:2), azole antifungal agents (fluconazole or miconazole) were administered intravenously. In 9 (81.8%) of the 11 evaluable cases, the antifungal chemotherapy was effective and the titers decreased to less than or equal to the previous titers in all cases. In 2 cases, the antifungal chemotherapy was not effective, and the titers did not decrease. These results suggest that serial Candida antigen detection provides a useful method in the early diagnosis of invasive candidiasis in febrile neutropenia and in determining the timing of the initiation of early antifungal chemotherapy. This method might also be useful in preventing the excess use of antifungal agents; thus preventing the proliferation of azole-resistant Candida infection. PMID- 10846835 TI - Changes in microbial flora in neutropenic patients with hematological disorders after the Hanshin-Awaji earthquake. AB - The earthquake that struck the Hanshin-Awaji district on January 17, 1995, was one of the most destructive to occur in Japan. We analyzed the changes in microorganisms isolated from patients with hematological disorders in relation to this earthquake. We reviewed a total of 4137 microbial cultures obtained over 1 year periods before and after the earthquake. There were 123 neutropenic patients admitted during the study period (54 before the earthquake and 69 afterwards). No significant differences in clinical characteristics and underlying diseases were found between the 2 groups. Polynomial analysis revealed a significant increase in the isolation of Candida albicans, Candida glabrata, and Pseudomonas aeruginosa from sputum and stool cultures after the earthquake. These microorganisms increased markedly in patients from severe earthquake-damaged areas. In neutropenic patients with hematological disorders, microbial isolates changed significantly after the Hanshin-Awaji earthquake. These changes may have been related to physical and emotional stress, as well as to the deterioration of environmental factors such as food, air, and water. PMID- 10846836 TI - Chronic graft-versus-host disease in children and adolescents after bone marrow transplantation from HLA-matched donors. AB - We analyzed 98 pediatric patients who underwent bone marrow transplantation (BMT) from serologically HLA-matched related donors (RD) or unrelated donors (UD) at our institute to clarify the actual status of chronic graft-versus-host disease (cGVHD). There were 36 evaluable cases of RD-BMT and 35 of UD-BMT. cGVHD was observed in 8 RD-BMT cases (22.2%) and in 23 UD-BMT cases (65.7%). In the RD-BMT cases, the limited and extensive types of cGVHD were observed in 4 cases each, whereas in the UD-BMT cases, the limited type was seen in 11 cases and the extensive type in 12. Prior acute GVHD was observed in 6 RD-BMT cases and in 18 UD-BMT cases. Two RD-BMT patients with extensive type cGVHD died of relapse and cytomegalovirus infection, and 4 UD-BMT patients died because of bronchiolitis obliterans, fungal infection, liver failure, and multiple organ failure, respectively. The incidence of cGVHD in these pediatric patients was as high as that in adult patients when UD-BMT was performed. Some UD-BMT patients required long-term immunosuppressive therapy after BMT. These findings suggest that cGVHD is a serious problem in pediatric UD-BMT. Therefore, intensive prophylaxis and treatment of GVHD must always be performed after UD-BMT. PMID- 10846837 TI - Quality of life in adult patients after stem cell transplantation. AB - Many articles pertaining to quality of life (QOL) following stem cell transplantation have been published in the US and western Europe. However, since the actions of health insurance systems and overall cultural aspects are strongly associated with QOL, investigations into QOL should be carried out within all countries. Therefore, we have investigated the QOL of adult patients following stem cell transplantation at 31 hospitals in Japan. The survivors, who were surveyed by mail questionnaire, were 20 years or older at the time of this study. The underlying diseases were acute lymphoblastic leukemia, acute myelogenous leukemia, chronic myelogenous leukemia, non-Hodgkin's lymphoma, Hodgkin's disease, myelodysplastic syndrome, and multiple myeloma. Median age at the time of the study was 36 years, and median interval after transplantation was 35.3 months. Of 383 patients surveyed, 282 (73.6%) responded to the questionnaire. One hundred and ninety-two patients were treated with an allogeneic-related transplantation, 52 with allogeneic-unrelated, and 38 with an autologous transplantation. Our data revealed that the length of time since transplantation and the diagnosis of chronic GVHD were associated with QOL. When unrelated and related transplantation recipients were compared, ratings on relief from pain, stability in weight, and confidence in dealing with daily life were lower among unrelated transplantation patients. PMID- 10846838 TI - Adult T-cell leukemia successfully treated with allogeneic bone marrow transplantation. AB - Adult T-cell leukemia (ATL) is associated with human T-cell leukemia virus type 1 (HTLV-1) and is known to be a refractory disease of highly poor prognosis. We describe a case of ATL treated with allogeneic bone marrow transplantation (allo BMT). The allo-BMT successfully induced complete remission in the patient. Currently, at 24 months post BMT, there has been no evidence of minimal residual disease (MRD) detected by polymerase chain reaction (PCR) assay for the T-cell receptor gamma chain gene. By contrast, PCR analysis demonstrated the reappearance of the cells harboring the integrations of the HTLV-1 proviral DNA 9 months after the BMT. These findings may imply a reversion to the carrier state rather than the recurrence of the leukemia from the MRD. The clinical consequence of our case illustrates that allo-BMT is an effective therapy, at least for achieving longer disease-free survival in ATL. PMID- 10846839 TI - Developing our understanding of the hematopoietic system. AB - The symposium entitled "Developing Our Understanding of the Hematopoietic System" was held in the 61st annual meeting of the Japanese Society of Hematology. Six speakers presented their interesting original study. PMID- 10846840 TI - Health for all in the 21st century a policy framework for the WHO European region. AB - The HEALTH 21 policy for the European Region of WHO provides the framework for taking up the challenges of achieving better health by applying the best strategies that have emerged from the Region's collective experience during the past 10-15 years. The arguments contained within this new policy demonstrate the essential relationship between health, poverty and social cohesion. The policy adopted by the WHO Regional Committee for Europe sets targets and outlines strategies that countries, organisation and civil society can use to turn national policies into practical operational programmes at local level through the European Region. PMID- 10846841 TI - Problems in diagnosing occupational allergy to flour: results of allergologic screening in apprentice bakers. AB - Baker's respiratory allergy is reported as one of the most common forms of occupational allergy in many countries. At the first stage of a three-year study of risk factors of occupational allergy we investigated the symptoms reported, and the results of skin prick tests (SPT) to common and flour allergens in 461 current apprentice bakers from three different regions of Poland before starting occupational exposure. The occurrence of cough was reported by 14 subjects (3%), dyspnoea by 4 (0.87%), rhinitis by 18 (3.9%), conjunctivitis by 12 (2.6%) and skin symptoms by 12 subjects (2.6%). Generally, 48 subjects (10.41%) reported at least one symptom, which might suggest the existence of allergic disease. Positive SPT to at least one allergen was found in 99 subjects (21.5%), including 82 subjects (17.78%) with positive SPT to common allergens, 13 subjects (2.81%) to flour allergens and 4 subjects (0.88%) sensitised only to Dermatophagoides farinae. The statistical analysis revealed significant correlation between positive SPT to common allergens and reported rhinitis and conjunctivitis, and between positive SPT to occupational allergens and reported dyspnoea. Moreover, a significant correlation between positive SPT to D. pteronyssinus and D. farinae was found. In our opinion, the results of SPT should be very carefully examined, when diagnosing occupational allergy, as in some apprentice bakers positive results of SPT to flour allergens are found before vocational training. In all apprentice bakers, SPT to common and occupational allergens should be performed before starting occupational exposure. It would ensure the exclusion of subjects already sensitised to occupational allergens. Positive SPT to Dermatophagoides farinae has a very limited value in recognising baker's allergy. PMID- 10846842 TI - Asthma due to kerosene exposure: three case reports. AB - Three families comprising 6 adults and 3 children experienced prolonged exposure to kerosene vapour as a result of accidental domestic oil storage tank spills in 3 separate incidents. All were previously well but within 6 to 8 weeks, 3 children and one adult developed asthma which has persisted for more than 2 years. The 5 remaining adults developed upper and lower respiratory tract symptoms (but not asthma) and symptoms of conjunctival irritation. Levels of kerosene vapour were measured in all rooms in one household and were within accepted toxicological limits. Kerosene vapour is not considered toxic. Asthma caused by kerosene vapour is previously undescribed. PMID- 10846843 TI - The different effects of black and white Vietnamese Aodai folk costumes on rectal temperature and heart rate in women walking intermittently in hot and sunny environment. AB - The present study was undertaken to investigate whether there is any difference in the rectal temperature and heart rate between women wearing black or white Aodai folk costume and walking intermittently in natural sunlight. The experiment was performed in the field outside buildings in Hanoi, Vietnam, between May and June, 1998. The only difference was in the colour (black or white) of clothing other physical characteristics, like materials, thickness, weight and so on, were nearly identical. Air temperature was around 39 degrees C, globe temperature around 52 degrees C and sun radiation reached 1010 W.m-2. Eight young female students (aged 20 years) participated in the experiment. They sat quietly on a chair inside the building for the first 30 min. Then they walked for 20 min at their ordinary walking speed in the sun, and rested for 10 min in the shade. This schedule was repeated three times. The main results are summarised as follows: (1) Rectal temperature was significantly lower in the black than in the white Aodai; (2) Clothing microclimate temperature at frontal chest level was also significantly lower in the black Aodai; (3) Heart rate was significantly lower in the black than in the white Aodai; (4) Clothing surface and inside temperatures measured at frontal chest level were significantly higher in the black than in the white Aodai. These results strongly suggest that the black Aodai could reduce heat strain more effectively than the white one. The physiological mechanism may result from more effective ventilation between skin and clothing in the black Aodai, due to higher temperature inside and outside this garment. PMID- 10846844 TI - Occupational exposure to static, ELF, VF and VLF magnetic fields and immune parameters. AB - Despite the important role of the immune system in defending the body against infections and cancer, very few investigations have been undertaken to study possible effects of electromagnetic fields on human immunity. The aim of the present study was to examine the effect of occupational exposure on hospital personnel operating magnetic resonance tomographs and on industrial workers at induction heaters. In both categories of workplaces, magnetic flux densities exceeding Austrian exposure standards have been registered. Because of the complexity and high redundancy of the immune system, an extensive range of assay systems was applied: relative and absolute numbers of lymphocytic subsets were counted, the proliferative activity of T and B cells determined, the production of interleukin 2, interferon gamma and tumour necrosis factor alpha analysed, serum immunoglobulins evaluated, as well as non-specific immunity of monocytes and granulocytes measured by their oxidative burst. The number of natural killer cells and oxidative burst in monocytes showed statistically significant differences in workers at induction heaters and controls. The observed effect on oxidative burst was counteracted by a higher number of active cells in the exposed group, indicating normal non-specific immunity. The high number of natural killer cells, observed in some of the study subjects working at induction heaters, was reconfirmed in another investigation and deserves a further follow up. PMID- 10846845 TI - Impact of new highways on road deaths: a case study in risk assessment. AB - Epidemiologists have generally avoided to assess risk for road deaths from high speed highways. We examined the validity of the claim that the Trans-Israel Highway, a six-lane 320 km toll road with higher design speed, and raised speed limits (120 kph), will reduce road deaths. We used models showing that death tolls vary with the fourth power of rise in driving speed. Risk assessments was derived from estimates of increase in the highway-induced traffic, the impact of higher speed limits (from 110 to 120 kph) and the so-called spillover effect from speed habituation. We predict a large rise in the number of killed or injured, even if the death risks per vkm is low on the Highway itself. With the Trans Israel Highway, death tolls--some 550 fatalities per year in 1995, could rise to as high as 900-1000 per year in 2010. Congestion produced by induced traffic will partially offset these effects. By contrast, death tolls from alternative strategies based on sustainable transportation policies could be reduced to less than 300 deaths per year. Risk assessment based on explicitly defined assumptions predicts high death tolls from the nationwide impact of raised speed on the Highway and its connecting roads. There is a need for new frameworks which impose the Code of Helsinki type requirements for the assessment and authorization of social decisions with adverse public health impacts. PMID- 10846846 TI - Concentrations of anaesthetic gases in hospital operating theatres. AB - Occupational exposure to anaesthetic gases (halothane, forane and nitrous oxide) was assessed in hospitals located in Lodz and its satellite towns. Individual dosimetry and stationary sampling methods were employed. The samples of air from workplaces were analysed by gas chromatography with mass detection or flow ionisation (halothane, forane) and by infra-red spectroscopy method (nitrous oxide). The concentrations of halothane and accompanying substances (ethanol, isopropanol and diethyl ether) indicate that Polish OELs were met in the majority of the hospitals. As Polish hygiene standards for forane and nitrous oxide are no available, the concentration values were compared with Swedish and German OELs. The comparison revealed that forane concentrations did not exceed Swedish OEL but nitrous oxide did exceed German maximum allowable levels. PMID- 10846847 TI - Changes in electrocortical arousal following acute trimethylbenzene administration in rats. AB - The purpose of this investigation was to compare the neurotoxic potential of trimethylbenzene (TMB) isomers (the solvents) with that of benzene derivatives with a smaller number of methyl groups (toluene). The experiments were performed on WAG/Rij rats with EEG recording electrodes implanted in the fronto-parietal cortex. The solvents, toluene or TMB isomers: 1,3,5-TMB (mesitylene), 1,2,3-TMB (hemimellitene) or 1,2,4-TMB (pseudocumene), were diluted with olive oil and administered intragastrically via gavage at an acute dose of 0.002, 0.008, or 0.032 mol/kg. The electrocortical activity was recorded for 20 min before, and for 60 min after the solvent administration. The electrocorticograms were analysed with respect to the number and duration of the high-voltage spindles (HVS), a form of activity sensitive to the arousal level. In case of each solvent the observed effect--inhibition of the HVS activity--was dose-related. However, the effect produced by TMB isomers was in each case less pronounced than that of toluene. Among TMBs, pseudocumene displayed the least significant effect, and the efficacy of two other TMB isomers was similar. PMID- 10846848 TI - Robert Graves and the origins of Irish medical journalism. PMID- 10846849 TI - A cure for cancer? Dealing with minimal residual disease. PMID- 10846850 TI - Lessons from sudden infant death syndrome. PMID- 10846851 TI - Pregnancy in Irish renal transplant recipients in the cyclosporine era. AB - BACKGROUND: The effect of renal transplantation on pregnancy in Irish women not receiving CyA has been reported previously. AIM: To examine all pregnancies occurring in Irish female renal transplant recipients since the introduction of CyA. METHODS: Using a community based approach, we identified 29 pregnancies in 19 women, aged between 16 and 45, mean age 30.3 years. RESULTS: These pregnancies ended in four miscarriages (13%), two intra-uterine deaths (6.9%) and 23 live births (79.3%). Of these live births, 73.9% were premature (< or = 36 weeks) and 65.2% were of low birth weight (< 2500 g). Admission to the neonatal intensive care was necessary in 61%, and two babies (8.7%) died in the neonatal period. Mean gestational age was 34 weeks, and mean birth weight was 2190 g. There was no change in graft function during pregnancy, with a small rise in serum creatinine post-partum (+9.64 mumol/L). The renal graft failed in three women (15.8%) by the end of the follow-up period. Compared with the precyclosporine era, the live birth rate was higher (79.3% versus 58%) with a trend towards lower birth weight and shorter gestation. CONCLUSION: Renal transplantation with CyA use is not a contraindication to pregnancy, but it is associated with increased risk, especially when the serum creatinine is > 175 mumol/L. PMID- 10846852 TI - The provision of a semen cryopreservation service for male cancer sufferers in the Republic of Ireland. AB - BACKGROUND: Chemotherapy, radiotherapy and surgery for cancer all carry risks of sterility. Semen cryopreservation would allow procreational ability to be preserved. METHODS: A human-assisted reproduction unit was set up in the Rotunda Hospital where semen was cryopreserved and stored for use. RESULTS: Semen was frozen for procreational potential during 1998 from 58 males about to have oncology therapy likely to render them sterile. The planning for this service and the modus operandi now in place are described. While cryopreservation was not unsuccessful in any case, substandard samples were obtained from 14 men. Pre freeze viral tests tested positive for CMV in six patients. One pregnancy is already on-going utilising IVF with thawed semen. One patient's death has been notified and the frozen semen disposed of. CONCLUSION: The service is, at present, outside public purse provision. It is hoped that in the near future this will change, as the deficit between charges and costs cannot be sustained in house forever. PMID- 10846853 TI - Initial experiences of a testicular sperm extraction programme for assisted reproduction in Ireland. AB - BACKGROUND: The technique aspirating spermatozoa from the testis is a new development in male infertility treatment. It is appropriate for infertile couples where the male has azoospermia but is still producing live motile spermatozoa in the testes. AIM: To describe the initial experiences of a testicular sperm extraction programme (TESE) coupled with intracytoplasmic sperm injection (ICSI) in 18 men during 1998. METHODS: Spermatozoa were obtained by direct aspiration from the testes using a 16 gauge needle with cannula and negative suction under local anaesthetic. All samples obtained were to be cryopreserved for use in a subsequent ICSI cycle. RESULTS: All five men with congenital bilateral absence of the vas deferens to be carriers of cystic fibrosis gene mutations. No gene deletions were found in their wives. No other cyto or molecular genetic abnormalities were otherwise found. Twenty one procedures were carried out. The mean number of aspirations was 1.72. Eleven samples from 10 men had sperm suitable for ICSI post-freeze. Post-procedure pain was the universal side-effect. Eight couples had a single attempt at ICSI, two couples two each. Fertilisation rate was 71.4%. Two pregnancies were achieved. CONCLUSION: TESE may give hope in selected cases of azoospermia of fathering a child without the involvement of a third party. PMID- 10846854 TI - Hospital-based stroke care in Ireland: results from one regional register. AB - BACKGROUND: Most patients with acute stroke are admitted to hospital. If stroke services in this country are to be improved, we need accurate and reliable information about the types of stroke patients being admitted, their present management and outcome. AIMS: To examine the demography, severity, level of investigation, length of stay, mortality and discharge location of prospectively identified consecutive stroke admissions to three general hospitals in South East Dublin. RESULTS: Three hundred and twenty nine consecutive stroke admissions to three general hospitals in South East Dublin were registered using the European Stroke Database over 50 weeks. The mean age was 73.3 years, whilst 20.1% patients were under 65 years. Prior to admission, 90% of patients were community dwelling with 14.9% of patients being dependent in activities of daily living. 22.4% of patients had some depression in level of consciousness on admission. The overall mortality rate was 26.1% whilst 136 (41.3%) were discharged home, 50 (15.2%) went to institutional care and 45 (13.7%) went to non general hospitals secondary rehabilitation units. The mean length of stay was 31.3 days. The combined poor outcome measure (mortality plus percentage of patients discharged to institutional care), was lower in one hospital compared to the other two hospitals (29.3% versus 44.65%, p > or = 0.05) probably reflecting case mix. Stroke accounted for 4.2% of all bed days in the major general hospital in this area. The overall CT scan rate was 84.5%, with 18.2% of CT scans showing a haemorrhagic component and two patients (0.8%) having brain tumours. Carotid doppler examinations were carried out in 37% of patients. CONCLUSION: The results demonstrate the high mortality and prolonged hospital stay for stroke patients in this area and emphasise the need for co-ordinated stroke care and regular audit to ensure most effective use of hospital resources. PMID- 10846855 TI - Evaluation of an alternative model of anticoagulant care. AB - BACKGROUND: Alternative models for the delivery of anticoagulant care are required in view of an estimated increase of 20% per annum in patients requiring treatment, and an inability for the present service to deal with this increase. This led to a restructuring of the anticoagulant service, which included the appointment of a nurse specialist and the implementation of a computer-assisted warfarin dosing system. AIM: To assess the impact of changes in the delivery of anticoagulant care in one unit. METHODS: A previous consultant-led delivery service of anticoagulant care was compared with the current nurse-led service. Two six month periods was compared. The end points of the study were INR control, appointment intervals and clinic size. A patient questionnaire was completed to assess patient satisfaction and education. RESULTS: In two years, attendance at walk-in clinics increased by 43% but the number attending anticoagulant clinics decreased by 50%. There was a parallel increase in the number of patients presenting on a flexible 'walk-in' basis for phlebotomy with postal dosing with 76% of patients now being managed on a flexible postal system. Anticoagulant control and reattendance intervals compare favourably, the percentage of patients attending at the maximal reattendance interval of eight weeks has increased from 3% to 15%. Patient knowledge and satisfaction scores were high on the questionnaire. Financially, this model of care is more cost effective than employing further doctors. CONCLUSION: The introduction of a nurse specialist managed service has allowed us to accommodate a 21% annual increase in patient numbers while improving the overall quality, efficiency and cost-effectiveness of the service and patient care. PMID- 10846856 TI - Feasibility of day case varicose vein surgery in a district general hospital. AB - BACKGROUND: Day case surgery may reduce inpatient bed requirements, and varicose surgery may be an ideal operation to test the functioning of a day surgery service. AIMS: To evaluate retrospectively the feasibility of day case varicose vein surgery in all-comers, and to identify the risk factors for admission. METHODS: Over a three-year period from July 1995 to July 1998, all patients requiring varicose vein surgery had their procedure performed as a day case. A standard technique of sapheno-femoral ligation with below-knee stripping of the long saphenous vein and multiple stab avulsions of varicosities was performed. All limbs were dressed with wool in crepe bandage and were reviewed post operatively at six weeks. RESULTS: Five hundred and forty two patients underwent varicose vein surgery, of whom 26% had bilateral varicose veins, 88% had primary varicose veins and 22% had recurrent disease. The procedure was performed under general anaesthesia in 86% of cases and under spinal anaesthesia in the rest. There was no peri-operative mortality. Ten patients (1.9%) developed post operative complications. Seventy two per cent of patients were discharged on the day of surgery, a further 25% required admission for one night due to minor anaesthetic complications and patient preference. Four per cent required admission for more than 24 hours. The need for overnight admission was associated with the age of the patients (p < 0.0001), bilateral varicose vein surgery (p < 0.005) and the use of spinal anaesthesia (p < 0.01). CONCLUSION: Although varicose vein surgery is safe, acceptable and cost-effective as a day case procedure, if attempted in all-comers, overnight admission will be required in a significant proportion. Pre-operative selection is advised. PMID- 10846857 TI - Sudden infant death syndrome--are parents getting the message? AB - BACKGROUND: Factors that place an infant at increased risk of sudden infant death include the prone sleep position, overheating and parental smoking, while practices such as bottle-feeding, co-sleeping and the use of pacifiers remain controversial. Major publicity campaigns have been undertaken, which have included the distribution of printed material and extensive media coverage. AIMS: To examine if Irish parents follow the currently recommended childcare guidelines to reduce the risk of sudden infant death and to examine factors that may have impact on their acceptance. METHODS: A random selection of 197 infants from the Birth Register of the Eastern Health Board. Parents were interviewed and a semi structured survey questionnaire was completed. RESULTS: Forty one per cent of infants are still placed on their side to sleep, an inherently unstable position. First time parents are more likely to place their infants on their backs. Over 60% of infants are exposed to one or more adults smoking in the home despite parental knowledge of its association with sudden infant death syndrome (SIDS). Sixty eight per cent of infants are overwrapped at night and parental understanding of what constitutes overwrapping is poor. Thirteen per cent of infants regularly co-sleep with their parents and 20% of these parents smoke. Pacifier use is common. CONCLUSION: Future programmes should target first time parents, should provide clear information regarding appropriate infant thermal environment, and should ensure regular updating of medical personnel so that they can instruct families on best current practice. Smoking remains a significant health issue with an impact on sudden infant death. PMID- 10846858 TI - Inner-city hospital closures in Dublin: implications for general practice. AB - BACKGROUND: The Meath, Adelaide and National Children's Hospitals provided more than 300,000 ambulatory care episodes in 1996, half to residents of the adjacent inner-city postal districts. With the closure of the group in 1998, alternative care arrangements must now be provided. AIMS: This paper examines the socio economic and primary care characteristics of the communities most affected. METHODS: Data on the 13 District Electoral Divisions surrounding the hospitals are drawn from a range of published sources. RESULTS: The populations affected include 52% in social classes 5 to 7, have an average Standardised Mortality Ratio of 122, have high levels of GMS eligibility and include special care issues such as drug dependency. A total of 38 GPs cover the area. Other markers of deprivation are summarised. CONCLUSION: The areas most affected by the closures have existing high levels of deprivation and social and medical need. Initiatives to respond to the closures must address these needs. PMID- 10846859 TI - The pattern of perforating eye injuries in Ireland. AB - BACKGROUND: Almost all perforating eye injuries present to dedicated eye units. Data from eye units are therefore reliable indicators of the incidence of perforating injuries. AIMS: To establish the aetiology and visual outcome of perforating eye injuries presenting to one unit over a twelve-month period. METHODS: Patients with eye injury were identified from the operating room register. Case-notes were reviewed and eye examinations were performed to determine current level of vision. RESULTS: A total of 37 perforating eye injuries were operated on in the 12-month period from January to December 1995. Thirty one patients (83.8%) were male. The mean age was 33.9 years. The chief causes were work related or do-it-yourself (DIY) in 32.4%, miscellaneous accidents in 27.0%, assault in 16.2%, road traffic accidents in 10.8%. Twenty eyes (54.1%) were blind with acuity < 6/60 at their most recent review. CONCLUSION: Work-related and DIY injuries are the most common causes of eye perforation. Many of these injuries are easily preventable with appropriate eye protection. The proportion of injuries resulting from road traffic accidents was low. Injuries due to assault are becoming more prevalent and carry a poor visual prognosis. PMID- 10846860 TI - Apolipoprotein E gene polymorphisms and serum cholesterol in healthy Irish adults: a proposed genetic marker for coronary artery disease risk. AB - BACKGROUND: The apolipoprotein (Apo) E gene, and thus its gene product, plays a central and pervasive role in lipid metabolism by serving as a ligand for lipoprotein receptors. Polymorphisms of this gene have been associated with variation in lipid phenotypes in some Caucasian and Asian populations, but not in others. No such study has been carried out in a resident Irish population. AIM: A study was designed to examine the relationship between serum cholesterols and Apo E genotype in a cohort of healthy Irish adults. METHODS: One hundred healthy Irish adults, aged 19-65 years, were recruited from the Cork City area. Two fasting blood samples were collected from each subject. One was assayed for serum cholesterols--total and low-density lipoprotein (LDL), and high-density lipoprotein (HDL)--while the other sample was used for isolation of genomic DNA and determination of Apo E genotype. RESULTS: While the E2 (12%) was the least prevalent, E3 was the most prevalent Apo E genotype (66%) in this group of healthy Irish adults. A significant Apo E gene-dosage effect was evident, whereby individuals with the Apo E2 genotype had a lower level of total cholesterol, E3 had intermediate levels, and E4 had a higher level. Moreover, those with the Apo E4 genotype had a significantly higher level of LDL cholesterol compared to E2 or E3 genotypes. There was no significant difference in mean serum adjusted HDL cholesterol levels between the three Apo E genotypes. CONCLUSION: These findings suggest that healthy Irish adults with the Apo E4 genotype have higher serum total and LDL-cholesterol levels than those with E2 or E3 Apo E genotypes and therefore may have a higher risk of atherosclerotic coronary artery disease and coronary heart disease in later life. PMID- 10846861 TI - Circadian rhythm of granulocyte-macrophage colony-stimulating factor in normal subjects and neutropenic hospitalised patients. AB - BACKGROUND: Granulocyte-macrophage colony-stimulating factor (GM-CSF), one of the haemopoietic growth factors, has rarely been detected in human serum. It has, therefore, been suggested that a paracrine model can explain its behaviour where the substance is produced and acts locally. An alternative explanation might be due to blood sampling time with GM-CSF concentrations undetectable at the nadir of secretion. HYPOTHESIS: We hypothesised that endogenous production of GM-CSF in humans is subject to diurnal rhythm. METHODS: Blood samples were obtained from 17 healthy individuals and 17 neutropenic hospitalised patients with haematological malignancies on myelosuppressive therapy at 6, 12, 18 and 24 hours. In the neutropenic patients, samples were collected at the nadir of the neutrophil count (ANC < 0.2 x 109/L). Serum was assayed for GM-CSF levels using an enzyme-linked immunosorbent assay method. RESULTS: There were significant differences in the mean levels of GM-CSF within the two groups (P < 0.001). In normal subjects, peak GM-CSF levels were reached at six hours (mean = 10.1 pg/ml). Peak levels were reached in hospitalised neutropenic patients at 18 hours (mean = 13.7 pg/ml). The difference between the peak GM-CSF levels in the two groups was not significant (P = 0.11). On factorial design analysis, there was a significant interaction between the time of blood collection and the subject groups (P < 0.001). CONCLUSIONS: Our data are consistent with a diurnal secretion pattern for GM-CSF in both normal and neutropenic patients. As this finding might have practical implications, including timing of administration of GM-CSF in neutropenic patients, further studies are suggested. PMID- 10846862 TI - A comparison of papaveretum-promethazine with morphine-ondansetron for patient controlled analgesia. AB - BACKGROUND: Patient-controlled analgesia (PCA) with intravenous morphine is commonplace. Antiemetics are often added to reduce side-effects. In our institution an unusual combination of papaveretum-promethazine is in use. AIMS: We set out to compare this combination with a more commonly used one (morphine ondansetron) by auditing the records of our pain-control team assessing analgesia and control of side-effects. METHODS: The records of two groups of 100 consecutive patients were studied. Pain scores were recorded 24 hours post operatively on a scale of zero to ten. Also recorded were sedation scores, pruritus, emesis, and usage of breakthrough analgesia. The groups contained the work of a variety of anaesthetists and surgeons, though the majority were orthopaedic cases. All results were recorded by one pain-control nurse, using the same question, over a nine-month period. RESULTS: Sedation scores were identical, while side-effects were few and similar in both groups. The use of breakthrough analgesia was also similar. However, the median pain-score of zero in the papaveretum group was significantly lower than that of two in the morphine group, (p < 0.001, Mann-Whitney U). CONCLUSION: Papaveretum-promethazine appeared to be effective for PCA. This combination is also much less expensive than morphine ondansetron. A previously described synergistic interaction between promethazine and opiates may be a factor in its success. Further evaluation of these drugs in PCA is warranted. PMID- 10846864 TI - Pharmacoeconomics in Ireland. Concepts and terminology. PMID- 10846865 TI - Successful management of penetrating cardiac trauma in a regional hospital. PMID- 10846863 TI - The effects of blood on rapid urease testing for Helicobacter pylori in mucosal biopsies from the gastric antrum. AB - BACKGROUND: While the eradication of Helicobacter pylori in patients with bleeding peptic ulcer disease (PUD) decreases the rate of ulcer re-bleeding, the sensitivity of the rapid urease test (RUT) for H. pylori diagnosis is lower in this setting. The aim of this study was therefore to determine if exposing a gastric biopsy specimen to blood before its use in the RUT (CLOtest) could account for these findings. METHODS: In patients undergoing endoscopy for the evaluation of dyspepsia gastric mucosal biopsies were obtained for H. pylori diagnosis (RUT, microbiology, and histology). Mucosal biopsies from each patient were also exposed to blood for 15, 30 and 45 minutes before use in the RUT. RESULTS: Using a combination of diagnostic tests (histology, microbiology and routine CLOtest) as the 'gold standard', the sensitivity, specificity, positive predictive value and negative predictive value of the CLOtest remained above 90% despite prior exposure of the gastric biopsy specimen to blood, and these values were not significantly different from the performance characteristics of the CLOtest processed in a routine manner. CONCLUSION: The exposure of gastric mucosal biopsy specimens to blood alone is not the explanation for the reduced sensitivity of the RUT in patients with bleeding peptic ulcers. PMID- 10846866 TI - Endometriosis in a scarless abdominal wall with underlying umbilical hernia. PMID- 10846867 TI - Bernardino Ramazzini and his treatise of the diseases of tradesmen. PMID- 10846868 TI - Pens and needles: the physician as writer. PMID- 10846869 TI - Mild cognitive impairment: transition between aging and Alzheimer's disease. AB - The concept of the boundary between normal aging and early Alzheimer's disease is a focus of a great deal of research in the field of aging and dementia. Presumably there is a continuum of function between normality and the earliest signs of Alzheimer's disease. This transitional condition has been labeled mild cognitive impairment. Mild cognitive impairment refers to individuals who have a memory impairment greater than what one would expect for age, yet general cognitive function is preserved. Similarly activities of daily living are normal. However, the memory function of these individuals is abnormal for age and education. These subjects do not meet criteria for Alzheimer's disease. When mild cognitive impairment subjects are followed longitudinally, they tend to convert to clinically probable Alzheimer's disease as a rate of 10-15% per year. This is in contrast to normal elderly subjects who will develop Alzheimer's disease at a rate of 1-2% per year. Certain predictor variables are available to determine which subjects are more likely to progress at a rapid rate. Mild cognitive impairment is an important topic for research in aging and dementia and has also become the subject of several multicenter treatment trials. PMID- 10846870 TI - [Sleep and respiratory disorders in myotonic dystrophy of Steinert]. AB - BACKGROUND: It has been hypothesized that hypersomnia and sleep related respiratory impairment are both central in origin in myotonic dystrophy. OBJECTIVE: To describe by means of video-polysomnographic recordings the central origin of the sleep respiratory disorders. PATIENTS AND METHODS: We studied 11 patients, 6 men and 5 women (mean age 42.7 years) with myotonic dystrophy. A moderate to severe ventilatory impairment of a primarily restrictive type was seen in all patients, three of them after the first episode of respiratory insufficiency. The patients were evaluated in order to determine their body mass index and presence of sleep-related complaints. Video-polysomnographic recordings (EEG, EOG, EKG, submental and tibialis anterior EMGs, respiration and Sa02) and pulmonary function tests were performed in each patient. Identical recordings were repeated in six cases, which were to undergo non-invasive bi-level ventilation (BiPAP) in order to adjust the inspiratory and expiratory pressures and the machine mode. RESULTS: We found slight hypopnea and apnea, predominantly of a central type, in stage 1 and REM sleep and alveolar hypoventilation in all patients. Sleep was disrupted and the efficiency index was very low. In three patients HLA typing showed a positive DQ6 haplotype. Six patients were treated with n-BiPAP. CONCLUSION: Nasal-BIPAP should be considered as an alternative in ventilatory support during sleep in these patients and video-polysomnography as a valid method of evaluating the ideal time to start treatment. PMID- 10846871 TI - [Drop attacks in patients with partial epilepsy]. AB - BACKGROUND: It is known the presence of sudden falls or epileptic drop-attacks (DA) in patients with partial epilepsy. OBJECTIVE: To review the clinical and electroencephalographic manifestations in our patients with partial epilepsy and DA. PATIENTS AND METHODS: Fifteen patients (9 males/6 females) over 18 years with partial epilepsy and epileptic falls were selected. RESULTS: The mean age was 39 years (24-56 years). The mean age at seizure onset were 13 years (8 months-49 years) for partial seizures and 26 years (2-54 years) for DA. Secondary generalized or not, all patients had complex partial seizures, associated with simple partial seizures in five (34%). All were treated with politherapy, but a good control was not achieved. EEG recordings showed frontal focus in 7, temporal focus in 8, secondary bilateral synchrony in 9, and increase of electroencephalographic abnormalities during sleep in 9. Cognitive and emotional disorders were observed in 8 and 6 patients, respectively. Nine patients suffered from status epilepticus. The causal lesions were connatal encephalopathy in 8 and criptogenic in the other 7. The main consequence of DA was recurrent craneal trauma in 9 patients. CONCLUSIONS: The presence of DA is considered an ominous change in the evolution of a partial epilepsy. It's associated with mental deterioration and emotional disturbances and with drug resistance. PMID- 10846872 TI - [Paraneoplastic neurological syndromes: from the diagnosis of exclusion to the use of immunological and molecular markers]. AB - For many years paraneoplastic neurological syndromes have been identified through the exclusion of other neurological complications in patients with cancer. The discovery that many paraneoplastic syndromes are associated with immunological reactions allows a specific and comprehensive definition of these disorders, which often can be promptly recognized by the serological detection of antineuronal antibodies. In a significant number of paraneoplastic syndromes the genes coding for the target onconeuronal antigens have been cloned and their functions are being elucidated. This study reviews the pathogenic mechanisms of paraneoplastic neurological syndromes and addresses the most frequently asked questions regarding their diagnosis and treatment. PMID- 10846873 TI - [Dural arteriovenous fistula of the brain]. PMID- 10846874 TI - [Posttraumatic intention tremor. Report of a case with a lesion in the mesencephalic area]. AB - We present a child who suffered severe coma because of an automovile accident remaining two months in coma with brain stem dysfunction and one and a half additional months until he was able to recognize people. After recovering, he showed severe kinetic tremor in the upper right extremity and discrete right hemiparesis as well as a moderate clinical cerebellar disfunction. Magnetic resonance (MR) study, performed 1 and 3 years after the coma, showed normal images of the cerebellum and thalami, while it disclosed signal changes in the mesencephalic region next to both red nuclei, being more severe in the right side. It is suggested that the kinetic tremor can be related with location of the lesion near red nucleus. PMID- 10846875 TI - [Autosomal recessive chorea-acanthocytosis linked to 9q21]. AB - A 34-year-old male, son of consanguineous parents, had a progressive neurological illness characterized by seizures, tics, choreic movements and mood changes. Acanthocytosis was present in blood. The level of creatine kinase was elevated. Normobetalipoproteinemia was noted. No KX group changes of McLeod syndrome were found. Serial neuroimaging studies demonstrated progressive caudate atrophy. Muscular biopsy confirmed the existence of non-specific myopathy. Genetic study demonstrated homozigosity for the 9q21 region. PMID- 10846876 TI - [Calcified cerebral metastases. Study of two cases and review of literature]. AB - Although the most frequent cause of intracranial neoplasms are metastases, they very rarely calcify; classic histopathological series have shown the incidence to be approximately 1%. We have found 54 reported cases of intracranial calcified metastasis, the most frequent primary tumour reported being pulmonary adenocarcinoma (13 cases) and adenocarcinoma of gastrointestinal tract and adenocarcinoma of breast (8 cases each). Some reports have shown a longer survival in patients with this kind of image on cerebral scans. We present two patients with intracranial calcified metastasis due to dissemination in both cases of pulmonary adenocarcinomas. The survival under treatment in both patients was similar to that of patients with no calcified metastasis on brain scans. So, facing calcified intracranial lesions, metastatic origin should be considered in the differential diagnosis, pulmonary tumours being the first to be ruled out. PMID- 10846877 TI - [Papillary fibroelastoma of the heart: a rare case of stroke]. PMID- 10846878 TI - [Deficiency neuropathies]. PMID- 10846879 TI - [Campylobacter coli-related meningitis in an adult male]. PMID- 10846880 TI - [Intracranial hypertension in a patient with pseudomigraine with pleocytosis]. PMID- 10846881 TI - [McArdle disease: a disease with high allele heterogeneity with different physiopathologic mechanism]. PMID- 10846882 TI - [Semont maneuver and vestibular rehabilitation exercises in the treatment of benign paroxysmal postural vertigo. A comparative study]. AB - BACKGROUND: Benign paraoxysmal postural vertigo (BPPV) is one of the most frequent causes of peripheral dizziness. Treatment based on vestibular rehabilitation exercises (VRE) is effective in 90% of the cases in an interval of 3 to 4 weeks. This treatment however is often abandoned by the patient. The only therapeutic maneuvers (based on otolyte release) are equally effective but present a high medium term recurrence. AIM: To compare the efficacy of the Semont maneuver (SM), the VRE and the combination of both in the treatment of BPPV at three months of follow up. MATERIAL AND METHODS: Forty patients with the clinical and electronystagmographic diagnosis of BPPV were divided into three groups: 1. SM, 2. VRE and 3. SM + VRE. The results were compared in regard to the signs and symptoms observed in the three groups of patients at 15 days, one month and at three months of treatment. RESULTS: The SM was effective in 80% of the patients at 15 days of treatment versus 45% of those receiving only VRE. The third group of patients, receiving the combined treatment, demonstrated a cure in 100% of the cases when evaluated at three months while only 66% of the SM group were found to be asymptomatic at the same time period (p < 0.05). CONCLUSIONS: The SM is easy to perform, rapid and effective in the short term, however has high recurrence. The VRE are effective in the long term based on the patient persistence. Combination of the two treatment leads to symptom remission in 100% of the patients at three months of treatment. PMID- 10846883 TI - Brainstem lesions decrease heart rate variability. AB - OBJECTIVE: Brainstem is an essential region in the regulation of the autonomic cardiovascular functions. Some authors have reported that medulla oblongata is very important in this regulation, but probably is not the unique because the sympathetic cardiovascular centers are spread in the brainstem. Since some years ago, we are able to measure heart rate variability which is a useful tool for assessing quantitatively both sympathetic and parasympathetic autonomic effects on the heart. The objective is to evaluate the effects of isolated brainstem lesion (IBL) on sympathetic and parasympathetic cardiovascular regulatory functions. PATIENTS AND METHODS: We analyzed the heart rate variability in time and frequency domains in 14 IBL patients and 25 age and sex matched healthy control subjects, measured from 24-h electrocardiogram. RESULTS: Low frequency power (0.001) and low frequency/high frequency power (0.05) were significantly higher in control subjects independently, all together or in subgroups (medulla oblongata affected or not). High frequency power (0.05) were higher too in control subjects. Variability in time domain (0.001 and 0.01) was lower when the medulla oblongata was affected. CONCLUSIONS: These results confirm that brainstem lesions cause cardiovascular autonomic dysregulation overall when the medulla oblongata is affected. The importance of this region in parasympathetic and sympathetic cardiovascular functions is showed. However, pontine and mesencephalic lesions seem to provoke a decrease only in sympathetic regulation. PMID- 10846884 TI - Drug treatment of HIV associated neuropsychiatric syndromes. AB - Psychotropic drugs are frequently used to treat the wide range of neuropsychiatric syndromes that patients infected with the human immunodeficiency virus (HIV) may develop. In order to administer these agents properly, physicians should take into account, among other factors, that: the central nervous system (CNS) of these patients is often impaired; they tend to suffer from one or more physical disorders; and they may be taking various other medications. The present paper reviews the clinical features and the general guidelines for administering neuroleptics, antidepressants, psychostimulants, benzodiazepines, opiates, lithium and carbamazepine in this group of patients, based on the literature and the authors' own clinical experience. PMID- 10846885 TI - [Anomalies of the common oculomotor nerve in ophthalmoplegic migraine]. PMID- 10846886 TI - Whipple's disease with isolated central nervous system symptomatology diagnosed by molecular identification of Tropheryma whippelii in peripheral blood. AB - We report a new case of Whipple's disease (WD) confined to the central nervous system. The patient presented with ataxia, ophthalmoplegia, hypersomnia, hemiparesis and generalized myorhythmia. The diagnosis was confirmed by identification of specific sequences of the causal agent of WD, the actinobacteria Tropheryma whippelii (TW), by PCR of DNA extracted from peripheral blood. An epidemiological survey of TW in patients with dementia suggests that WD is an uncommon cause of dementia in our population. Molecular methods may allow rapid identification of TW in peripheral fluids, and non-invasive diagnosis of this disorder. PMID- 10846887 TI - [Recurrent familial brachial plexopathy as the only clinical expression of neuropathy with susceptibility to pressure]. AB - We report a family with hereditary neuropathy with liability to pressure palsies (HNPP) and chromosome 17p11.2 deletion. This family exhibits a peculiar phenotype consisting in recurrent brachial plexopathy episodes. This phenotype has to be distinguished from hereditary neuralgic amyotrophy on clinical grounds. Although the incidence of brachial plexopathy on HNPP is relatively high it is unusual as the sole symptom of the disease. It is noteworthy that in the six published families with this peculiar phenotype most of the acute episodes became evident after sleep. A greater liability of the plexus and a greater vulnerability to mechanical factors during sleep hours are the suggested mechanisms to explain this rare clinical onset. Recurrent painless brachial plexopathy when associated to generalized conduction abnormalities should suggest a HNPP. PMID- 10846888 TI - Hypoglossal schwannoma: an uncommon cause of twelfth-nerve palsy. AB - Hypoglossal schwannoma is an uncommon cause of 12th nerve palsy. We here report a pathologically proven case of left 12th nerve schwannoma in a 56-year-old woman who presented with progressive left tongue hemiatrophy and in whom cranial MRI disclosed a mass near the foramen jugular. Surgery showed a mass involving the hypoglossal nerve and the pathological examination was diagnostic of schwannoma. PMID- 10846889 TI - [Comment on the article "Cerebral arteriopathy with subcortical infarctions and leucoencephalopathy with dominant autosomal inheritance (CADASIL). Clinical study and morphology"]. PMID- 10846890 TI - [Observation on the classification of primary headaches]. PMID- 10846891 TI - [Diagnosis of temporary arteritis and training in headaches in primary health care]. PMID- 10846892 TI - [Cough syncope with transient neurological deficit: case report]. PMID- 10846893 TI - [Training in dietetics and nutrition from the point of view of the primary care physician]. AB - BACKGROUND AND GOALS: Professionals in primary health care have to face a large number of patients with pathologies arising out of nutritional and dietary disorders as well as cope with society's growing interest in such issues. For this reason, we have attempted to assess the extent of the knowledge in questions of nutrition and dietetics that primary health care doctors feel they have received in comparison with what they might have considered necessary, as well as assess the capacity that these doctors feel they have to cope with clinical situations requiring a knowledge of nutrition. SCOPE AND SUBJECTS: 250 doctors working in Primary Health Care and belonging to the Tarragona Province Medical Association. ACTION: Participants received a self-administered questionnaire in which they had to: a) answer closed questions on their sense of clinical ability to handle dietary and nutritional problems; b) give a score for the importance that 62 previously-defined subjects should have in their general medical training; and c) give a score for the attention paid to these topics during their undergraduate training. RESULTS: 36 doctors (age: 38.6 +/- 10 years) with an average accumulated experience in medicine of 12.5 +/- 9.2 years replied to the survey. Respondents reported that 42.5 +/- 25% of their patients required nutritional or dietary action and only 28 +/- 24% receive the same. As for their undergraduate training in nutrition, 19.4% of them considered it to be non existent and 58.4% described it as insufficient. Respondents reported little ability to handle different clinical situations, particularly involving the identification of patients requiring secondary vitamin therapy or nutritional support and the prescription of low lactose diets. Except for the area of biochemistry and nutritional physiology, respondents reported a great discrepancy between the attention that should have been given during their undergraduate years to the subjects proposed and the training they really received, particularly in questions of clinical dietetics, diet and the prevention of disease as well as nutritional support in disease. CONCLUSIONS: The undergraduate training in questions of dietetics and nutrition is clearly seen to be scant according to primary health care physicians. This might explain in part the insufficient clinical capacity that these professionals feel they have in such matters and the high percentage of patients who do not receive the nutritional and dietary attention these doctors feel is necessary. PMID- 10846894 TI - Dietary intake in adolescents from south-east Spain and its relationship with physical activity. AB - Dietary intake (energy, macronutrients and micronutrients) and its possible relationship with levels of physical activity were studied in an adolescent group consisting of 142 boys and 197 girls aged between 14-18 years, from the region of Murcia. A continuous, 7 day dietetic register was used, and the population was stratified by physical activity (low, medium and high) and by sex. Results show an excess of energy in the diets of the male group, concurring with tricipital fold measurements, which were higher than the mean value for the Spanish adolescent population. A significant negative relationship was found (P < 0.05) between levels of physical activity and body fat content in the female group. Lipid excess and carbohydrates deficit were the principal dietetic imbalances found. Micronutrient deficits were observed in vitamin E, iron, zinc and magnesium. Although the study does not reveal significant differences in the alimentary habits of active and sedentary adolescents, the beneficial effects of exercise can be considered to partly compensate the dietetic imbalances found. PMID- 10846895 TI - [Dietary-therapeutic product supply in unitary doses at a university hospital: effect on consumption and costs]. AB - GOALS: To analyze the consumption and costs of the Enteral Nutrition products (diets and dietary supplements) dispensed from the Nutrition unit in a 735-bed general hospital during two six-month periods, before and after the implementation of a unit-dose (UD) distribution system. MATERIALS AND METHODS: The defined daily dose (DDD) methodology was used. A DDD of 1,500 Kcal/day was defined for complete diets in adults, with 1,000 Kcal/day for complete diets in children, 1,200 Kcal/day for "start-up" diets, 500 Kcal/day for normocaloric supplements and 600 Kcal/day for hypercaloric supplements and 450 Kcal for very low caloire diets. RESULTS: The total consumption in the hospital fell from 10.21 DDD per 100 stays/day (DED in its Spanish acronym), representing generated costs of 8,640,130 pesetas between November, 1997, and April, 1998, to 6.25 DED in the same period for 1998-99, with costs amounting to 6,674,775 pesetas (-23%). In the services handled using UD, the total consumption declined from 9.3 DED between November, 1997, and April, 1998, to 5.2 DED in the same period for 1998-99. The consumption of standard/special diets and dietary supplements went from 1.38/0.76/7.16 DED to 1.48/0.53/3.19 DED during the two periods under study, with an accompanying drop in costs of -1,806,598 pesetas (-30%). In percentage terms, the standard diets varied from 26% to 41% (of the total cost generated in the services with UD), special diets from 28% to 22% and dietary supplements from 54% to 37%. In those services where the UD was not applied (ICU and reanimation), the total consumption of diets remained stable with a slight drop from 25.3 DED to 24.3 DED. CONCLUSIONS: The unit dose system has improved the prescription of therapeutic dietary products in our hospital and has reduced the costs, mainly through the notable decrease in the dispensation of supplements. PMID- 10846896 TI - [Quality of the home parenteral nutrition program: 14 years of experience at a general university hospital]. AB - The quality of home parenteral nutrition (NPD in its Spanish acronym) depends on the frequency and type of complication associated with NPD treatment and the likelihood of survival. The present study assesses the quality of the NPD programme in place in our hospital in terms of survival, infections and mechanical complications. A retrospective study was carried out into the clinical follow-up data of all the patients (n = 24) included in our NPD programme since its start in 1985 until 1998 (14 years). An estimate is made for: a) the annual index of infectious complications (IAC in its Spanish acronym), b) the annual index of mechanical complications (MAC in its Spanish acronym) and c) the likelihood of survival by means of the Kaplan-Meier method. The quality specifications adopted are those of the literature reflecting the current provision of NPD programmes and the survival values of patients undergoing dialysis for chronic kidney failure. The most frequent pathology in our context is benign (70.8%), distributed as follows: small bowel syndrome of ischaemic origin (45.8%), small bowel syndrome of non-ischaemic origin (12.5%) and idiopathic intestinal pseudo-obstruction (12.5%). The patients with benign pathologies present a higher survival rate than patients with neoplastic disease (95% in the fifth year of treatment versus 45% at twenty months), with a statistically significant difference. The annual index of infectious complications is 0.6 (median value of the 14 years studied). Similarly, the annual indices of obstructions and thromboses are 0.11 and 0.0095, respectively. In our opinion, the quality of the NPD programme in place at our hospital is highly satisfactory because both the survival rate and the annual indices of mechanical and infectious complications are acceptable with regard to the programmes in place in the international sphere. In addition, in terms of survival, NPD seems slightly more effective than dialysis for chronic kidney disease. PMID- 10846897 TI - [Prescription and costs of artificial nutrition before and after a new utilization protocol]. AB - In order to improve the quality of prescription and reduce expenditure, the artificial nutrition department has designed a new usage protocol: justification sheet, table of indications for parenteral nutrition (PN), nutritional assessment, etc. The Pharmacy Department is in charge of the follow-up of this programme and maintaining the database derived from it. Comparing the data for the year before (1996) and after (1998) implementation of this programme, we can see that as a result 9 patients/day switched from total parenteral nutrition by central administration to hypocaloric peripheral nutrition, with the prevalence of the former dropping to 1.4% of admissions and 2% of stays. The share of enteral nutrition (30.6%) and hypocaloric peripheral nutrition (27.8%) increases in the overall expenditure on artificial nutrition, with a reduction in the total parenteral nutrition (41.6%). The costs of parenteral nutrition per admission and stay are reduced by 39.3% and 33.5%, respectively, to 1,625 and 183 pesetas. This represents a saving of 16.5 million pesetas. The expenditure on PN elements declines by two percentage points to 2.2% of pharmaceutical expenditure. As an additional benefit, it is possible to identify each of the clinical situations which give rise to this indication and to allocate costs by diagnosis and department. Together with the containment of expenditure, it has been possible to increase the participation of everyone in the rational use of clinical nutrition thus highlighting its interdisciplinary nature. PMID- 10846898 TI - [Assessment of dietary intake in institutionalized elderly patients with dysphagia]. AB - Elderly institutionalised patients are at risk from the nutritional point of view. This fact is even more important in those with swallowing difficulties. To avoid the immediate consequences an inadequate nutrition has in older people's quality of life and prognosis is vital to achieve an adequate diet with a strict control of their food and fluid intake. PMID- 10846899 TI - [The tissue specificity of the protective action of cytoplasmic factors on the membrane-bound system of Ca2+ transport in the sarcoplasmic reticulum of the heart and the skeletal muscles]. AB - Whether soluble cytoplasmic factors (SCF) can protect the membrane-associated Ca2+ transport system in sarcoplasmic reticulum (SR) was studied in controls, stress and adaptation to it. Ca-transport was damaged in stress much more in the skeletal muscles compared to the heart. Initial performance of Ca-pump in the skeletal muscles was decreased by 43%, in the heart--no change. After stress, thermoresistance of the enzyme in the skeletal muscles was 3-5 times less than in the myocardium. The enzyme reaction on adaptation and stress in both tissues is the same--a significant rise in Ca-pump thermoresistance. When preadapted animals are exposed to stress, deceleration of the initial speed and resistance of the enzyme in the myocardium are fully prevented while in the skeletal muscles Ca pump resistance is higher than in the controls. In the heart, SCF have a protective effect on thermoresistance. In the skeletal muscles, SCF protect initial speed of the enzyme in the control, in stress and adaptation to it. A cross-over protective SCF (from adapted animals) effect was found on the stress impaired membrane system of Ca-transport. Adapted SCF elevate 2.5-fold thermoresistance of the enzyme. Correlation between cytoplasmic and membrane protective mechanisms in stress and adaptation to it is considered. PMID- 10846900 TI - [Nitric oxide and lipid peroxidation as factors in endogenous intoxication in emergency states]. AB - K coefficient which is an integral indicator of the unbalance in the system lipid peroxidation--antioxidant system (LPO-AOS) and content of nitrite, nitric oxide end product, were followed up in 31 patients with concomitant brain injury, in 24 patients with burn injury and 72 patients in hypertensive crisis. Coefficient K was estimated by serum levels of dienic conjugates, malonic dialdehyde, lipid peroxidation degree, alpha-tocopherol, ceruloplasmin on the disease day 1, 3, 7 and 14. Nitrite serum levels rose by day 14 in all the patients and were the highest in hypertensive crisis patients. Coefficient K was significantly elevated in all the patients compared to controls being maximal in patients with concomitant brain injury. Thus, activation of LPO and biosynthesis of nitric oxide contribute much to endogenic intoxication in patients on critical care. PMID- 10846901 TI - [The weightlessness disease problem]. PMID- 10846902 TI - [Carnosine stabilization of the normal erythrocyte membranes and in experimental diabetes]. AB - The effect of carnosine on red blood cell membranes was studied in the in vitro and in vivo experiments. Carnosine (70-720 microM) is shown to protect healthy donors' red blood cells from acidic hemolysis increasing population of stable red blood cells and postponing time to maximal rate of hemolysis. It was demonstrated on the model of streptozotocin-induced diabetes in rats that carnosine treatment (50 mg/kg/day during 7 days, per os) prevents a decrease in the hemolytic stability of red blood cells in diabetic animals. The findings suggest usefulness of carnosine as a possible treatment of diabetes patients. PMID- 10846903 TI - [The effect of laser hemotherapy on the acid glycosaminoglycan content of the liver and blood plasma in rats in the dynamics of an alcoholic coma]. AB - The aim of the study was determination of the content of acid glycosaminoglicans (GAG) in hepatic tissue and blood plasma of rats in the course of alcoholic coma and in response to laser irradiation of venous blood. The alcoholic intoxication was achieved by introduction of 40% ethanol through a metal probe into the stomach. The content of acid GAG was defined according to Diche. Alcoholic coma caused reduction of GAG levels in hepatic tissue and an increase in blood plasma which were correlated with the severity of neurological state. Laser irradiation of venous blood promoted a trend to normalization of GAG content in the investigated tissues. A relative reduction in GAG plasma levels after laser hemotherapy indicates effectiveness of the conducted therapy. PMID- 10846904 TI - [The dependence of the connective tissue reaction to stress on typological body characteristics]. AB - The experiment on 52 Wistar rats was made to study dependence of connective tissue reactions on typological properties of the body. The type of the reaction is closely related with the degree of changes in connective tissue structures of the digestive organs (periodontium, stomach) and the lungs. Acute stress stimulates degradation of the noncollagen proteins of the connective tissue main substance. PMID- 10846905 TI - [The anti-inflammatory activity of intal and beta-carotene in a model of experimental granulomatous lung inflammation]. AB - A-25 sephadex-induced granulomatous inflammation of the lungs in rats was treated with beta-carotene and intal in inhalations. Both drugs showed antiinflammatory activity reducing the area of alveolitis and emphysema in the lungs, number of neutrophils and lymphocytes in the bronchoalveolar fluid. The experimental data allow to recommend further trial of intal and beta-carotene in granulomatous pulmonary diseases. PMID- 10846906 TI - [The excretion of ascorbic acid and its metabolites in uremia and hemodialysis]. AB - Renal excretion of ascorbic, dehydroascorbic and diketogulonic acids in uremia and relevant loss in hemodialysis are measured in comparison with those in patients with uremic syndrome (prior to hemodialysis) and in healthy subjects (control). Renal elimination of ascorbic acid was higher while of dehydroascorbic acid lower vs control. Elimination of diketogulonic acid was similar to control. In a session of hemodialysis, the organism loses 132.0 +/- 13.6 mg of ascorbic, 132.0 +/- 10.0 mg of dehydroascorbic and 204.0 +/- 9.0 mg of diketogulonic acid. 48-hour urinary losses of the patients reached 8.4 +/- 1.4, 19.6 +/- 1.1, 75.6 +/ 1.5 mg, respectively. Compared to control, hemodialysis patients lose the above acids 24.3, 2.7 and 4.6 times more. PMID- 10846907 TI - [The stress phenomenon. Emotional stress and its role in pathology]. PMID- 10846908 TI - [Perioperative arterial hypertension. A pending problem]. PMID- 10846909 TI - [In Process Citation] PMID- 10846910 TI - [Cardiovascular response to tracheal intubation in patients with intracranial tumor. Comparative study between urapidil and lidocaine]. AB - OBJECTIVES: To compare the effects of urapidil and lidocaine on cardiovascular response to laryngoscopy and tracheal intubation in patients with brain tumors. PATIENTS AND METHODS: Prospective, randomized double-blind study of 40 ASA II-III patients undergoing elective surgery for supratentorial tumor resection. The patients were assigned to two groups of 20 to receive an intravenous bolus of 1.5 mg/kg of lidocaine or 0.4 mg/kg urapidil before laryngoscopy and intubation. Anesthetic induction was performed with 0.03 mg/kg midazolam, 3 micrograms/kg of fentanyl, 5 mg/kg of thiopental and 0.2 mg/kg of vecuronium. Anesthesia was maintained with N2O/O2 (60%/40%) and isoflurane (0.5% expired). The following variables were recorded: mean blood pressure, heart rate and arterial oxygen saturation (SpO2) at baseline and 1, 2 and 3 min after induction and at 1, 2, 3, 4, 5 and 10 min after laryngoscopy and tracheal intubation. RESULTS: The two groups were comparable with respect to age, sex, weight, height and baseline hemodynamics. No statistically significant differences in hemodynamic variables were found between the two groups. Mean blood pressure in the postintubation period stayed near baseline and heart rate increased significantly after laryngoscopy and tracheal intubation in both groups. SpO2 decreased 7 min after administration of urapidil and stayed lower throughout the study period and was statistically different from SpO2 in the lidocaine group. All changes described were within clinically normal ranges. CONCLUSIONS: In patients undergoing neurosurgery to remove a supratentorial tumor, both lidocaine and urapidil reduce the pressor response to laryngoscopy and tracheal intubation at the doses given in this study. However, neither was able to prevent the increase in heart rate. PMID- 10846911 TI - [Study of presurgical anxiety in urologic, gynecologic, and ophthalmologic surgery as a function of the administration or non-administration of anxiolytic premedication]. AB - OBJECTIVE: Anxiety commonly appears before surgery, triggering a set of events that can be described as a general stress response. We measured changes in preoperative stress in relation to premedication or not with diazepam in patients undergoing urologic, gynecologic and ophthalmologic surgery. PATIENTS AND METHOD: We enrolled 87 patients undergoing elective surgery in urology (n = 30), gynecology (n = 30) and ophthalmology (n = 27). Randomly, the patients were premedicated with diazepam, placebo or nothing. During the preoperative visit the patient was administered a test to determine the level of anxiety and blood pressure, heart rate and potassium plasma concentration (K+) the night before surgery and moments before entering the operating theater. RESULTS: No significant differences in anxiety between patients undergoing different types of surgery were detected in the operating theater; however, at that time, anxiety was found to be significantly less among patients premedicated with diazepam than among those given placebo. Preoperative anxiety was significantly less in ophthalmology patients and in those premedicated with diazepam. Both systolic and diastolic blood pressure increased significantly among urologic and ophthalmologic surgery patients and in those who were not premedicated with diazepam regardless of type of surgery. Heart rate did not change significantly. Potassemia decreased significantly in all patients. CONCLUSION: The discrepancy between the psychological and somatic response to the stress conditions studied may be the result of the calming effect of the preoperative visit by the anesthesiologist and to premedication with a tranquilizer. However, neither the preoperative visit nor premedication seem to completely block the stress response. PMID- 10846912 TI - [Direct effect of anesthetic inducers on the electromyographic signals of the adductor pollicis]. AB - OBJECTIVES: To analyze the effect of thiopental, propofol, etomidate and midazolam on electromyographic study of the adductor pollicis before muscle relaxation. PATIENTS AND METHOD: Prospective study of 96 adults ASA I-II patients (gynecologic surgery) by electromyographic (Reflexograph Datex) recording of the adductor pollicis every 20 seconds with train-of-four (TOF) stimuli. After giving informed consent, the patients were randomized for type of induction agent. We studied changes in T1 (amplitude of the first response to TOF) and the TOF-ratio (T4/T1). The induction agent (thiopental 5 mg/kg), propofol 3 mg/kg, midazolam 0.2 mg/kg or etomidate 0.3 mg/kg) was given by slow 30 s injection 100 s after calibration of the Relaxograph. Changes in T1 and the TOF-ratio were recorded until 240 s. RESULTS: The groups were statistically homogeneous. In the thiopental group, T1 readings at 20, 40, 60 and 80 s (control times) were statistically different from the readings at 180, 220 and 240 s (after administering thiopental). T1 was significantly different at 120 and 200 s in the etomidate group. CONCLUSIONS: Thiopental and etomidate induce slight muscle relaxation at the expense of a reduction in T1 without changing the TOF-ratio, an effect that might be useful for rapid induction. Propofol and midazolam do not affect the electromyogram of the adductor pollicis. Thus, propofol is the anesthetic induction agent of choice for clinical studies on non-depolarizing muscle relaxants given that it changes neither T1 nor the TOF-ratio. PMID- 10846913 TI - [Ketorolac versus tramadol: comparative study of analgesic efficacy in the postoperative pain in abdominal hysterectomy]. AB - OBJECTIVE: To compare the analgesic efficacy of tramadol to that of ketorolac trometamol administered intravenously and at fixed times over the 24 hours after abdominal hysterectomy. PATIENTS AND METHODS: This controlled, double blind, randomized clinical trial enrolled 76 women undergoing abdominal hysterectomies. Two treatment groups were formed: the TRA (Tramadol) group received 100 mg and the KET (Ketorolac) group 30 mg administered every 6 hours intravenously. Patients were ASA I-II and aged 35 to 65 years old. Patients were excluded from the study if hysterectomy was performed because of a tumor, or if there was a history of bleeding dyscrasias, of gastric or duodenal ulcers or of allergy to drugs in the study or if other analgesics had been used within 15 days of surgery. Analgesic efficacy was assessed using a visual analog scale (VAS) and a verbal response scale (VRS). Need for top-up analgesia was recorded, as were the number of patients withdrawing before the end of the study and the side effects attributable to treatment. RESULTS: The mean VAS score throughout the study was 3.6 for the TRA group and 4.4 for the KET group (non-significant, p = 0.05). Likewise, VRS scores were similar. In the first 12 h after surgery, VAS scores in the TRA group were statistically lower than those of the KET group (p < 0.05). Nine patients abandoned the study before it ended: 3 in group TRA and 1 in the KET group. Three withdrew in the TRA group (2 because of vomiting and 1 for administrative reasons). Six left the KET group (4 for uncontrolled pain, 1 for impossible intubation and 1 for administrative reasons. In the TRA group, 38% experienced vomiting whereas only 8% did so in the KET group. No serious postoperative complications were recorded. CONCLUSIONS: During the first 12 hours following surgery, a 100 mg dose of tramadol has been shown to provide more effective pain relief than 30 mg of ketorolac administered intravenously every 6 hours. The only drawback to administering the drug intravenously was the high incidence of postoperative vomiting. PMID- 10846914 TI - [Usefulness of meperidine in anesthesiology]. AB - Meperidine was the first synthetic opioid agent. It acts mainly as an antagonist of mu (#m) receptors and has an analgesic potency ten times greater than that of morphine. Like other opioid drugs, meperidine causes nausea, vomiting, urinary retention and respiratory depression; a point of difference, however, is that it acts on nerve fibers and has properties similar to those of local anesthetics. It has therefore been used as an alternative to other opioids for anesthetic blockade. We review the indications and contraindications of meperidine administered by different routes. For pain, epidural administration has proven to be a good alternative to intravenous administration and epidural meperidine has been combined with local anesthetics using lower doses of both drugs and producing fewer side effects. Intradural meperidine has been used as the sole anesthetic agent in various types of surgery, its principal advantage being that it provides long-lasting postoperative analgesia. Spinal meperidine has the advantage over morphine of a lower incidence of respiratory depression, particularly late-occurring depression. An intravenous route has been used for treating moderate to severe pain, for regional anesthesia, for premedication and for analgesia during anesthesia. Meperidine's action on kappa receptors has meant that it is considered the most effective drug for treating postanesthetic trembling. Although meperidine has been used effectively to treat non-surgical pain, mainly from colic, this review focuses on its usefulness in the perioperative period. PMID- 10846915 TI - [Hyperinsulinemic hypoglycemia in children. Anesthetic treatment]. PMID- 10846916 TI - [Anesthesia recovery with sevoflurane in children]. PMID- 10846917 TI - [General anesthesia with laryngeal mask and remifentanil for remodelling and corrective osteosynthesis of neuropathic foot in a case of type I Charcot-Marie Tooth disease]. PMID- 10846918 TI - [A new case of tracheal injury following difficult intubation]. PMID- 10846919 TI - [Anesthesia in a case of type I Arnold-Chiari malformation]. PMID- 10846920 TI - [Anesthetic treatment of a patient diagnosed with exercise-induced anaphylaxis and cholinergic urticaria]. PMID- 10846921 TI - [Gas embolism in orthopedic surgery of the leg]. PMID- 10846922 TI - [Enantiomers and local anesthetics]. PMID- 10846923 TI - [Influence of inhaled glucocorticoids on bone mineral density and bone metabolism]. AB - Inhaled glucocorticoids (IGs) are today the first-line treatment for bronchial asthma. The systemic effects of inhaled glucocorticoids, such as suppressing the hypothalamic-pituitary-adrenal axis, are generally less than those with oral glucocorticoids. However, there is a long-term risk of adverse effects on bone. The objective of this piece was to review the published data on the effects of IGs on bone metabolism markers and bone mineral density in adults and in pediatric patients. The reviewed studies do not provide uniform results. Nevertheless, in general they suggest that IGs can affect metabolism and bone mineral density, especially: 1) when high doses are administered (more than 400 micrograms/day in children and more than 800 micrograms/day in adults), 2) in pediatric patients, in whom growth in stature can also be affected, 3) in patients whose intake of calcium and vitamin D is inadequate, and 4) in postmenopausal women not undergoing hormone replacement therapy. In general, at therapeutically equivalent doses, beclomethasone has a greater deleterious effect on bone than does budesonide, which in turn has more of an effect than does fluticasone. In addition to the obvious precaution of using the lowest effective dose, other proposed preventive measures include: 1) adequate instruction on the use of aerosols, 2) the use of large volume spacer devices, 3) rinsing the mouth after administering IGs, and 4) dietary adjustments or supplements in order to ensure an adequate intake of calcium and vitamin D. PMID- 10846924 TI - Vaccine-associated paralytic poliomyelitis in Brazil, 1989-1995. AB - At the present time, the only poliovirus-caused poliomyelitis cases reported in Brazil and other countries of the Americas are of vaccine etiology. It is important for epidemiological surveillance and immunization programs to evaluate the epidemiological profile of cases of vaccine-associated paralytic poliomyelitis (VAPP) in order to establish criteria for case definition and vaccination strategies. To research VAPP in Brazil, 30 cases diagnosed and classified as such by the Ministry of Health between 1989 and 1995 were submitted to a descriptive study of clinical, laboratory, and epidemiological data. In addition, the risk of occurrence of VAPP was estimated in relation to determinants based on a cohort of 3,656 persons with acute flaccid paralysis. Among individuals who had received oral polio vaccine (OPV) from 4 to 40 days before the onset of paralysis, we found a relative risk of 8.88 (95% CI: 4.37 18.03) for VAPP as compared with persons who had not been vaccinated during the same time interval. For individuals who developed VAPP in the period following national vaccination days, the estimated relative risk was 2.94 (95% CI: 1.44 6.00). For the first dose of OPV administered to the general population the estimated risk was 1 case of VAPP for every 2.39 million doses; for total doses of OPV the risk was 1 case in 13.03 million doses. A major share of VAPP cases were related to children affected by prodromes (fever and gastrointestinal signs and/or symptoms), isolation of vaccine poliovirus type 2, paralysis of the lower limbs, and a mean age of 1 year. PMID- 10846925 TI - [Fecal antigens of Fasciola hepatica potentially useful in the diagnosis of fascioliasis]. AB - This study describes which antigens of Fasciola hepatica are present in the feces of patients with chronic fascioliasis and in the feces of rats infected experimentally with F. hepatica metacercariae. Using a Western blot assay technique with hyperimmune serum obtained from excretory-secretory antigens of adult F. hepatica, we found in the patients' feces antigens of possible diagnostic interest, with molecular weights of 14, 19, 20, 23, 25, 32, 46, 51, and 62 kilodaltons (kDa). In addition, we showed that the peptides of 14, 20, 23, and 51 kDa are also recognized by the majority of the sera from chronic patients. We used affinity chromatography to purify the antigens present in the feces of rats that had been infected for 6 to 12 weeks, using ES78 monoclonal antibody bound to CNBr-activated Sepharose 4B. Through that approach, we identified six polypeptides, of 11, 14, 26, 32, 47, and 51 kDa; three more polypeptides, of 17, 24, and 66 kDa, could only be identified in the feces of rats that had been infected for 10 to 12 weeks. Our results suggest that these polypeptides could be antigens common to both parasitic stages. This is particularly true for the polypeptides of 14, 24, 26, and 51 kDa, because they reacted with the immune sera, the human sera, and the ES78 monoclonal antibody. These polypeptides could be important markers for acute and chronic fascioliasis. PMID- 10846926 TI - [Tuberculosis recurrence and its risk factors]. AB - The persistence of tuberculosis bacilli in patients who are cured, thus causing recurrence, is an important issue. This case-control study investigated individual and institutional risk factors for relapse by analyzing independent variables related to the patient, the use of antituberculosis drugs, and the service delivered at health care institutions; 56 cases and 105 controls were interviewed. Recurrence was defined as a new tuberculosis episode after the patient had been successfully treated. Controls were selected from among patients who were treated and cured of pulmonary tuberculosis and who did not experience a relapse. Regression models were proposed to control confounding factors or effect modifiers. The variables identified as risk factors for relapse were those related to erratic patient behavior (missing medical appointments and therefore not picking up the medication, not taking the medication, taking the wrong dosage), age, and stress from life events; adverse reactions to antituberculosis drugs; and problems in the organization of health care services that resulted in patients receiving insufficient dosages or amounts of antituberculosis drugs. Receiving information regarding treatment duration provided protection against recurrence. The knowledge regarding these risk factors should result in more intensive follow-up and in more use of directly observed treatment of tuberculosis in order to prevent relapse. PMID- 10846927 TI - Urinary fluoride excretion by children 4-6 years old in a south Texas community. AB - This study evaluated urinary fluoride excretion by school children 4-6 years old who were living in a south Texas rural community that had concentrations of fluoride in drinking water supplies generally around the optimal level. We took supervised collections of urine samples in the morning and afternoon at school, and parents of the participating students collected nocturnal samples. We recorded the beginning and end times of the three collection periods and then determined the urinary volume and urinary flow for each of the periods. We measured urinary fluoride concentrations and calculated the urinary excretion rate per hour. The children had breakfast and lunch provided at the school, where the drinking water contained 1.0-1.3 milligrams/liter (mg/L) fluoride. Fluoride concentrations in the tested household water supplies, from wells, ranged from 0.1 to 3.2 mg/L fluoride. The children's average urinary fluoride concentrations found for the day were similar to those for the night, with means ranging from 1.26 mg/L to 1.42 mg/L. Average excretion was 36.4 micrograms/h in the morning, 45.6 micrograms/h in the afternoon, and 17.5 micrograms/h at night. The lower nocturnal excretion rates are easily explained by low urinary flow at night. Based on the 15 hours of urine collected, the extrapolated 24-hour fluoride excretion was 749 micrograms. In conjunction with similar studies, the data from this study will help in developing upper limits for urinary fluoride excretion that are appropriate for avoiding unsightly fluorosis while providing optimal protection against dental decay. PMID- 10846928 TI - [Analysis of participation in surveys in 5 countries: the importance for public health research]. AB - This study compares participation rates and reasons for nonresponse in surveys conducted in five countries of Latin America and the Caribbean. The objective of the surveys was to measure the prevalence of risk behaviors affecting the transmission of human immunodeficiency virus. The surveys were based on probability samples of the population of both sexes between 15 and 49 years old, except in Mexico, where only men were included. Proportions of three components of participation were estimated: residences interviewed, interviewed residences with eligible persons, and eligible persons who completed the interview. In addition, an overall index that combined the three components was calculated. The overall response rate ranged from 35.6% in Mexico to 81.4% in Chile. The component with the greatest variability was the participation of eligible persons, which ranged from 50% in Mexico to 95% in Cuba. These values were lower than what had been expected, especially among men, and will serve to guide future surveys, since rejection rates higher than the ones expected in the protocol should be considered. The results make it possible to infer the validity of the prevalence estimates for the various observed risk behaviors. The results also establish a benchmark to calculate the sample size in future surveys and to improve research methodology. PMID- 10846929 TI - [Validation of diagnostic scales of family functioning for use in primary health care services]. AB - Care provided to patients in primary health services should include both a physical and a psychological assessment. In many cases treatment success depends on understanding individuals and their interactions, especially within the family; it is important for health professionals to be trained in this area. We investigated the validity of three well-known diagnostic scales of family functioning from the United States of America (FACES III, Beavers-Timberlawn, and GARF) for use in our setting in Porto Alegre, Brazil. The three scales were used with 31 families and compared to the results from clinical evaluations (semistructured family interviews), which were considered the gold standard. FACES III was also used with a sample of 102 families in the community. The Beavers-Timberlawn and GARF scales were strongly and positively correlated with the clinical evaluations. In contrast, our results suggest that FACES III is not a useful screening tool in primary care to detect the risk of psychiatric disorders. The Beavers-Timberlawn and GARF instruments showed great usefulness for formulating and classifying diagnoses of family functioning. PMID- 10846930 TI - [Mineral composition of selected tissues and organs of white-tailed sea eagle]. AB - The analysis of the concentrations of K, S, P, Mg, Na, Ca, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, As, Rb, Sr, Y, Ag, Cd, In, Cs, La, Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb, Lu, Hg, Tl, Pb, Bi, Th and U in tissues and organs of white-tailed sea eagle collected dead in Poland has revealed that the metal of risk is lead, and to a somewhat degree also mercury. An intoxication of white-tailed sea eagles with lead is due to ingestion of lead pellets from the waterfowl injured or killed by the hunters, which than become recaptured by the birds. In the case of mercury a source of elevated concentrations of that element in tissues and organs of some white-tailed sea eagles examined is their food (waterfowl and fish) originating from a coast of the Baltic Sea and the Firth of Szczecin. PMID- 10846931 TI - [The contamination of white-tailed sea eagles with organochlorines in Poland]. AB - The residues of PCBs, DDTs, HCHs, HCBz, PCBz, CHLs, aldrin, dieldrin, endrin, isodrin, endosulfan 1, endosulfan 2, mirex, TCPM-H and TCPM-OH were determined in breast muscles and an egg of white-tailed sea eagles collected in Poland in 1991 1995. The method of measurement was capillary gas chromatography and low resolution mass spectrometry (HRGC/LRMS) after a non-destructive extraction, clean-up and fractionation of the sample. Only aldrin, endrin, endosulfan 1 and endosulfan 2 were absent in birds and egg examined. Some of the adult white tailed sea eagles collected dead from the coastal area of the Baltic Sea still remain relatively high contaminated with organochlorines, and the concentrations of PCBs and DDTs in those birds ranged between 2300-2600 and 490-2000 micrograms/g lipids, respectively. In dead egg concentration of PCBs was 390 micrograms/g lipids (25 micrograms/g wet weight), while of DDTs 270 micrograms/g lipids (18 micrograms/g w.w.). PMID- 10846932 TI - [Polychlorinated biphenyls and organochlorine pesticides in soils in Poland]. AB - The concentrations of organochlorine pesticides such as DDTs, HCHs, CHLs, HCBz and polychlorinated biphenyls (PCBs) were determined in 59 soil samples collected in Poland in 1990-1994 to understand the scale of contamination, spatial distribution and sources of these compounds. Identification and quantification of organochlorines were carried out using HRGC-ECD (63Ni) and HRGC-LRMS. All soil samples were contaminated by DDTs, PCBs and HCHs, while HCBz and CHLs were absent in a few samples. DDTs dominated as organochlorine residues in Polish soil and on the concentration of other insecticides was two orders of magnitude lower. Percentage composition of DDTs (about 60% of p,p'-DDT) indicates a very slow metabolism of DDT in Polish soils. Spatial distribution of HCHs in Poland is similar to distribution of DDTs, while HCBz and CHLs are rather evenly distributed. PCBs concentrations in soil of the Former Soviet Union Army Base are 10 times higher then in urban soils, and 100 times higher then in cultivated and forests soils. PMID- 10846933 TI - [Content of nitrates and nitrites in raw mild in different regions of the country]. AB - The content of nitrates and nitrites in raw milk in different regions of the country was studied. The results obtained dependent both on the factory from which the samples origin and on the seasons of the year. In 37% of samples there was insignificant increase in the level of nitrates, and in 31.9 of nitrites. PMID- 10846934 TI - [Content of nitrates and glycoalcaloids in mature tubers of Polish potato]. AB - Nitrates and glycoalcaloids (TGA) content in mature tubers of 29 potato table cultivars of different earliness were examined during 1992-1998. Excess of nitrates (more than 250 mg NaNO3/kg of product--the upper limit for food safety in Poland) was found in 24 tuber samples what makes 16% of total number (154) tested samples. Samples with the highest amounts of nitrates belonged mainly to very early and early cultivars. TGA concentrations lower than the upper limit TGA for food safety (200 mg/kg of product) were estimated in all 129 tested tuber samples. Only in 8 samples TGA content was between 100 and 140 mg/kg of product what could cause bitter taste. PMID- 10846935 TI - [Contents of radon 222Rn in drinking water of Sweradow Zdroj and Czerniawa Zdroj]. AB - Radon-222 concentration in surface water, wells and tap water in Swieradow Zdroj and in Czerniawa Zdroj has been quantitative determined. The measurements were performed using the alpha liquid scintillation counting method. Radon arithmetic mean for water of individual wells in Swieradow Zdroj was found to be 438.5 Bq/l within the range between 42 Bq/ and 1095 Bq/l. The appropriate mean value for water of individual wells Czerniawa Zdroj was 165 Bq/l within the range from 25.8 Bq/l and 402.4 Bq/l. Waterworks in Czerniawa Zdroj is supplied with the surface water in which the radon concentration is low (about 2 Bq/l). Water works in Swieradow Zdroj is supplied with the mixed water consisting from the surface water, which main characteristic is low radon concentration (below 11 Bq/l), and from artesian well and mine gallery water with the radon concentration from 321 Bq/l to 464 Bq/l in it. This water is mixed in various ratios and therefore the radon concentration in it was within 4 Bq/l and 79 Bq/l. PMID- 10846936 TI - [Campylobacter in surface waters for communal purposes in Bydgoszcz region]. AB - The aim of our work was to determine the frequency of Campylobacter occurrence in surface water taken from lakes and rivers. For this purpose the membrane filters technique in microaerophylic conditions and Columbia Agar with the antibiotics addition as the culture medium were used. The genera of Campylobacter were determined. At the same time the purity of water's samples was estimated. The total number of bacteria as well as the number of Coli spp. were determined. The results indicate that 33% of water samples were contaminated with Campylobacter (predominated C. coli and C. jejuni). The percentage increased with the degree of contamination with Coli. PMID- 10846937 TI - [The sorption of polycyclic aromatic hydrocarbons in Zegrzynskie Lake]. AB - The sorption of polycyclic aromatic hydrocarbons in aquatic sediments from Zegrzynskie Lake was examined. Batch experiment was performed in order to determine sorption efficiency in different kinds of sediments from Zegrzynskie Lake. Five polycyclic aromatic hydrocarbons (phenanthrene, fluoranthene, pyrene, chrysene benzo[a]pyrene) were chosen to this experiment and sorption process was examined on seven sediments of different properties. Chosen hydrocarbons are of different structure of molecule and different chemical and physical properties. Concentrations of polycyclic aromatic hydrocarbons in aquatic sediments and in water phase were measured in the following order: extraction with dichloromethane, concentration on rotary evaporator, silica gel clean up, n hexane elution, concentration on rotary evaporator and in vials, GC/MS analysis. Chemical composition of aquatic sediments were examined using methods for sewage sludge and soils analysis. In every sediment concentrations of PAHs, organic matter and organic: carbon, phosphorus, nitrogen and sulphur were measured. Also fractional analysis of sediments was made. Isotherms of sorption were measured for these sediments and compounds. Equations of these isotherms were performed and were used in order to find relationships between sorption efficiency and sediments composition. Depending on sediment properties and composition different concentrations of polycyclic aromatic hydrocarbons were found in solid phase. Sediments of high quantities of organic matter and small particles were the best sorbents for PAHs. Fluoranthene, pyrene and chrysene were efficiently sorbed in sediments of high concentration of organic matter. And efficiency of phenanhrene and benzo[a]pyrene sorption were better in sediments with high quantity of organic sulphur. PMID- 10846938 TI - [Application of the chromatographic-spectrophotometric method for the analytical control of l-ascorbic acid in pharmaceutical and plant material]. AB - The conditions of chromatographic separation allowing the isolation of L-ascorbic acid from its products of decomposition and other accompanying substances in the determined pharmaceutic preparation and plant materials were established. The isolated L-ascorbic acid was signed by the extractive--spectrophotometric method using coupled redox-complexation reactions with iron(III), 1, 10 phenantroline and bromophenol blue system. The analytical procedure allowing the evaluation of durability of vitamin C in multivitamin "Vitaral" preparate was described. The procedure was used also for the evaluation of the loss of L-ascorbic acid in the parsley and lovage in the process of drying and storage. PMID- 10846939 TI - [Current nutrition supporting programs of the population in the USA]. AB - In this work main programs of nutrition-supporting in the USA were presented as a part of national and local social and health policy. Following programs were reviewed namely: FSP (The Food Stamp Program), NSLP (The National School Lunch Program), SBP (School Breakfast Program), SMP (The Special Milk Program), WIC Program (The Special Supplemental Food For Women, Infants and Children), SFSP (The Summer Food Service Program). The level of monthly income per person (no more than 130% of poverty line) is the condition for participation in every program. Some programs (e.g. FSP) are addressed to all citizens, who are fulfilling income criteria, whereas other (e.g. WIC, SBP) are available to selected population groups only. Assistance within these programs may occur in various forms (e.g. food stamps for family, possibilities in using of low cost meals, free of cost milk for school children). Authors suggest to utilize some of US experience in nutrition-supporting programs also in Poland. PMID- 10846940 TI - [Oxidizing agents in hair cosmetic products and methods of their identification and determination]. AB - Hydrogen peroxide and oxidazing agents are used in hair lightening products, in oxidants of permanent wave products, hair dyeing products and toothpastes. In accordance with the Cosmetic Directive of the European Union (Council Directive of 27 July 1976 r. Annex III p. 12) maximum permitted concentration in the finished above mentioned cosmetic products is 12% in hair care preparations, 4% in skin-care preparations, 2% in nail hardening. The method identification and determination hydrogen peroxide and oxidazing agents is very useful for control cosmetic products on market. PMID- 10846941 TI - [Different possibilities and methods of realization of physical education in medium size primary schools]. AB - During spring '98 data about conditions and realization of physical education were based on a representative, national sample of 480 medium size (8-23 classes) primary schools. Information on sport rooms and areas, size of groups, teachers' qualification, place and type of lessons was collected. Only 35% of schools had a gymnasium. But nevertheless, a programme with 3 physical education classes a week was usually realized. Most of classes took place on sport areas (90% of schools had them). Up to 6% of classes was carried out on the corridor or in another inadequate place. The number of students taking part in training was often too large. Among younger children the subjects of classes were usually sport plays and games, while among elder students--they were usually ball games. The majority of teachers lacked physical education qualifications. Professional physical educationers worked usually in large urban schools that had a gymnasium, while unqualified teachers usually carried out physical education lessons in little rural schools that lacked a gymnasium. The data shows, that the lack of sport equipment was usually connected with low qualifications of the teachers. PMID- 10846942 TI - Trauma rounds. PMID- 10846943 TI - Tennessee Tar Wars. PMID- 10846944 TI - From research to policy: UT Center seeks to make a difference. PMID- 10846945 TI - Confusion in the ranks. PMID- 10846946 TI - A rational approach to the management of carpal tunnel syndrome in the workplace. AB - Carpal tunnel syndrome (CTS) occurring in the working population continues to be a frequent cause of unpleasant patient symptoms, loss of worker productivity, and substantial expense to employers. The cause of CTS in these patients is often unknown, but is frequently attributed to work activities. Current scientific literature suggests that CTS is usually a multifactorial process, with both patient and environmental contributors that should be considered in determining work-relatedness. Outcome following carpal tunnel surgery is dependent on a positive patient attitude, physician encouragement, and employer cooperation. Determinations of permanent impairment and work restrictions should be made by physicians experienced in caring for upper extremity conditions, and should be based on objective evidence of anatomic dysfunction. Legislative efforts regarding Workers' Compensation law should be undertaken to encourage patients towards a faster and more-complete recovery. PMID- 10846947 TI - An uncommon case of green urine. PMID- 10846949 TI - Tennessee legislation on safer medical devices. PMID- 10846948 TI - Development of thyroid follicular adenoma on simvastatin therapy. AB - HMG-CoA reductase inhibitors (lovastatin, simvastatin, fluvastatin, pravastatin) constitute a potent class of cholesterol-lowering agents, which are increasingly being used these days for primary and secondary prevention of atherosclerotic heart disease. Despite having good overall safety and efficacy profiles, these medications can still cause significant adverse effects including transient elevation of hepatic transaminases, myopathy, and rhabdomyolysis. Preclinical studies have demonstrated a potential of neoplasia in rats. However in clinical trials HMG-CoA reductase inhibitors have not been found to be neoplastic in humans. The dosage used in humans is also significantly lower and therefore it is expected to have a good safety margin. But this may not be entirely true considering the mechanism of neoplastic transformation, which is thought to be different in humans as compared to other species. We report a patient, who developed follicular adenoma with prominent Hurthle cell changes after being on simvastatin for three months but not during one year of pravastatin therapy. In elderly female patients with hyperlipidemia requiring pharmacologic treatment, especially those with a prior history of multinodular goiter, one should consider using an agent which has not been shown to cause thyroid tumors even in animal models. Patients should continue to be followed with frequent periodic thyroid palpation in addition to the usual biochemical monitoring required while on these agents. PMID- 10846950 TI - [Primary reconstruction in surgery for breast cancer?]. PMID- 10846951 TI - [Depression among the elderly. Focus on the diagnosis]. PMID- 10846952 TI - [Abnormal fetal ultrasound findings after maternal chickenpox infection]. AB - Chicken pox infection in the first and early in the second trimester can lead to intrauterine infection and a 2% risk of developing congenital varicella syndrome (CVS). CVS is characterized by one or more of the following malformations: hypoplasia of a limb, scarring of the skin, microcephaly, cataract, microophthalmia, cerebellar dysplasia, gastrointestinal malformations and urogenital malformations. Most of these malformations can be seen by ultrasound. Foetal viral infection can also lead to intrauterine growth retardation, polyhydramnios and hydrops foetalis. This paper is a review of the literature with the focus on ultrasound findings in CVS. The options for performing prenatal diagnostic tests and the consequences of maternal varicella zoster (chickenpox) for prenatal care are discussed. PMID- 10846953 TI - [Primary breast reconstruction in connection with mastectomy for breast cancer. Indication, procedure and immediate surgical results]. AB - Mastectomy and immediate reconstruction of 122 breasts were performed in 109 patients in close collaboration between plastic surgeons and general surgeons. In 56 patients reconstruction was performed using tissue expanders including 13 bilateral operations, 29 patients had a latissimus dorsi myocutaneous flap and 24 a free transverse rectus abdominis myocutaneous flap. There were 27 postoperative local complications in 122 reconstructions (22%), in five the reconstruction was lost. Only patients clinically in stage I were considered for reconstruction. After histopathological staging 27 patients received systemic treatment and 10 local radiotherapy as well. There was no complication during systemic therapy related to reconstruction. In 10 cases local radiotherapy was performed in full, with a delay of four weeks in one patient and a need for correction of the radiation field during treatment in one patient. PMID- 10846954 TI - [Transfusion-related hepatitis C. The Danish "look back" survey]. AB - This study accumulated results of the HCV lookback in Denmark and described the morbidity of the infected recipients. Donor records were identified for at least ten years back, and recipients still alive were tested for hepatitis C. Those with positive results were referred for clinical evaluation. A total of 150 Danish anti-HCV positive donors had donated blood to 1018 recipients of whom 288 (29%) were still alive. Because of age, malignancy or other severe diseases 118 (41%) of these were not contacted. Of 157 recipients screened for HCV, 128 (82%) were anti-HCV positive and 88 (56%) were HCV-RNA positive. Among the HCV-RNA positive recipients symptoms were present in 38% (25/66 reported), elevated ALT was found in 53% (41/77 tested) and cirrhosis was found in 11% (6/54 biopsied). Treatment with interferon-alpha was initiated in 23 patients, corresponding to 26% of HCV-RNA positive recipients. PMID- 10846955 TI - [Transportation of patients with acute myocardial infarction]. AB - The aim of this study was to describe the results and experiences from a local hospital concerning transports with patients with acute myocardial infarction to primary PTCA. Our material includes 27 patients who had primary PTCA. There were no complications during the transports. None of the patients had hypotension or arrythmia requiring treatment during the transports. One patient had ventricular fibrillation before the transport. None of the patients died during the transports. Two patients died in the period following PTCA. The transport delay was 1 hour and 40 minutes on average. Future randomized studies like DANAMI 2 will show if PTCA with delayed treatment, or immediate thrombolysis, give better results for the patient. PMID- 10846956 TI - [Epoxy in occupational environment--an old acquaintance in a new dress]. AB - This article describes a cluster of epoxy related contact dermatitis in a glassfibre reinforcement plant, using mostly preimpregnated epoxylaminate (Pre Preg), but also fluid epoxy-products. An occupational medical and dermatological examination revealed nine of 26 cases as allergic contact eczema, 14 as toxic epoxyrelated eczema and three cases of other non-occupationally related skin diseases. The plant uses seven different epoxyresins with 15 epoxyhardeners. Preventive measures were gradually improved, but even then it was difficult to rule the production into safe procedures with this potent allergen used in large scale production of windmill wings. PMID- 10846958 TI - [Chronic Chagas disease--an echo from youth]. AB - American trypanosomiasis (Chagas' disease) is uncommon in Europe, but occurs frequently in South and Central America where it causes major problems. A case is presented of a 57 year old woman born in Venezuela who showed signs of chronic Chagas' disease after living 32 years in Denmark. The epidemiology, modes of transmission, clinical manifestations, diagnosis and treatment of Chagas' disease are described. PMID- 10846957 TI - [Giant cell arteritis and scalp necrosis]. AB - We present a case of a 75-year old woman, who developed scalp necrosis as a result of giant cell arteritis in the temporal arteries. This is a very rare, but serious complication of temporal arteritis, which implies an increased risk of visual loss and other catastrophic sequelae. Healing of the ulceration was achieved after cortico-steroid therapy supplemented with split skin transplantation. PMID- 10846959 TI - [Negative bone scintigraphy in a child with acute osteomyelitis]. AB - Diagnosis of acute osteomyelitis in childhood is conventionally established by a combination of clinical symptoms and results of inflammation parameters in blood and blood culture, plain radiography, bone scintigraphy and ultrasound. High sensitivity of bone scintigraphy enables it to be positive in the majority of cases, especially when clinical symptoms and blood tests are inconclusive. We present a case of a nine month old girl with an established diagnosis of vertebral osteomyelitis. Her two bone scintigraphies were negative despite obvious destruction of vertebra L5 and S1, documented by plain radiography and magnetic resonance imaging (MRI). MRI should be used when conventional examinations are inconclusive. PMID- 10846960 TI - [Alibi of the researcher. A reflection on publications and lists of publications]. PMID- 10846961 TI - [Picture of the month. Mastocytosis]. PMID- 10846962 TI - [Spontaneous course of obsessive-compulsive condition (OCD)]. PMID- 10846963 TI - [Fear of anesthesia]. PMID- 10846964 TI - [Are younger physicians satisfied with their education?]. PMID- 10846965 TI - [Venous leg ulcers]. PMID- 10846966 TI - [Memory and pain]. PMID- 10846967 TI - Urinary incontinence. Impact on working women. AB - Until recently, the impact of urinary incontinence (UI) on working women, a population generally characterized as healthy, has not been the focus of research. Women employed full time at a large university center participated in a cross sectional survey about UI. Of the 1,113 women surveyed, age 18 and older, 21% (n = 232) reported UI at least monthly. Incontinent women were significantly older and had a higher body mass index than continent women. Using disposable products, limiting fluids, avoiding caffeinated beverages, using voiding schedules, and keeping extra clothes or underwear were strategies used to manage UI at work. Responses to an open ended question related to the impact of UI on working life included: interference with sleep and resulting fatigue at work, embarrassment, alteration of concentration, and emotional distress. Implications for nurses are discussed in relation to assessment, education, and management of UI in the occupational setting. PMID- 10846969 TI - Creating a successful occupational health and safety program. Using workers' perceptions. AB - Workers' perceptions of risk directly influence the behavioral choices they make. This directly impacts the type of safety precautions taken on the job. It is imperative that perceptions be considered when planning and implementing a worksite health and safety program. Occupational and environmental health nurses are in the perfect position to ensure workers' perceptions are considered, thus reducing worksite injury and illness and increasing company profits. PMID- 10846968 TI - Video technology. Use in nursing research. AB - Video technology is becoming more popular as a research tool because it has unique features that capture accurately and comprehensively the nature of nursing phenomena. Video technology is used extensively in nursing as an educational tool or intervention, a means of monitoring quality assurance standards, and as tool to collect research data. Videotaping is useful because is provides continuous multi-media, multisensory information about the subject and its context. Credibility for video data is based on the question of to what extent do the data or results adequately represent true behavior. The three main issues in video research that impact the credibility of data are observer bias, participant reactivity to knowledge of being videotaped, and maintenance of consistent data quality. Not all threats to credibility can be overcome. The key is to acknowledge insurmountable problems, to consider their limitations in data analysis, and to evaluate and report their effects. PMID- 10846970 TI - Legal aspects of work related stress in nursing. Exploring the issues. AB - Nurses are under a great deal of distress related to a variety of work stressors. Work related stress jeopardizes the mental and physical well being of nurses, as well as the quality of care provided for clients. The Occupational Safety and Health Act of 1970 holds employers responsible for the mental and physical well being of their employees. However, the regulations are not always straightforward concerning compensation claims for mental disabilities due to work stress. Recovery of claims is possible if standards (i.e., personal injury, causal relationship between injury and employment) are met. Requirements for recovery of physical/mental and mental/physical claims are agreed on by most states, whereas mental/mental claims are controversial among states and courts. Nurses can monitor and influence legislation to create requirements to encourage employers to develop preventive approaches to reducing work induced stress. PMID- 10846971 TI - Starting a business as a nurse consultant: practical considerations. AB - Nursing experience translates well to self employment in the occupational and environmental health field. However, nurses must conduct a self assessment to determine whether owning a business is a good fit for their personality and work style. Exploring which services to offer is the next step in starting a business and consists of determining not only which service to offer (e.g., writing policies or protocols, providing clinical services) but also which type of consultation model to use (i.e., purchase of expertise, doctor/client, process consultation). Every business must have a plan. A business plan is essential to starting a business. It solidifies the entrepreneur's focus, lays the foundation for the business, provides a tool for evaluating success, and is a strong tool for soliciting financial support. The name of the business is an important consideration because it is often the first contact the customers have with the occupational health nurse consultant. The name must be descriptive, appropriate, and memorable. PMID- 10846972 TI - Developing a strategic plan for the new millennium. PMID- 10846973 TI - Postexposure prophylaxis for HIV exposed health care workers. PMID- 10846974 TI - AHNA explores new direction. PMID- 10846975 TI - Working out stress. PMID- 10846977 TI - Getting started with e-mail--it's "easy" and "exciting". PMID- 10846978 TI - Financial empowerment: the holistic nurse entrepreneur. PMID- 10846976 TI - The world of personal and professional development. A different path to healing wisdom. PMID- 10846979 TI - To refer or not to refer. PMID- 10846981 TI - Clear seeing ahead. PMID- 10846980 TI - Manifesting our visions. PMID- 10846982 TI - The holistic nurse entrepreneur: the last taboo. PMID- 10846983 TI - Natural cycles. PMID- 10846984 TI - Hospital and home. AB - This paper addresses the hospital/community interface as an emerging context of health care practice. As a consequence of industry reforms health service managers are looking to the community space as a location for delivery of acute health care. This focus on the community is sharpened by the promise of cost savings and enhanced by the seemingly limitless potential of biomedical technology. The paper argues that the interface of hospital and community is a conceptual space where two different types of health services meet, bringing with them different cultural practices and expectations. The 'hospital in the home' programs that structure health care at this interface provide the delivery of acute nursing and medical care and the accoutrements of this care in the community, the neighbourhood, the home. Consequently, the home is becoming the new site for high technology 'hospital' care. This domestication of illness technology is contrasted with the notion of home as a place of sanctuary, familiarity and belonging. PMID- 10846985 TI - Revitalize to survive. AB - The important topic of the future of nursing directs members of the profession to consider that future critically. This paper advocates a future-building rather than a future-entering approach as nursing encounters the third millennium. That view is prompted by candid statements, gathered through a national survey of Australian nurse academics, which indicate that the term 'nursing in the future' could be an oxymoron. Discussion reports on and reflects upon deficits to systems of support for scholarly nursing, a current state of exhaustion amongst members of the profession and the perceived existence of inappropriate attitudes to professional responsibilities. The voices of study participants give rich suggestions for the way forward to professional revitalisation. Recommendations include adoption of new paradigms for nursing's identity and creation of means by which the voices of all nurses may be encouraged to participate in discourse concerning nursing's future in health care provision. PMID- 10846986 TI - Specialisation in nursing. A survey of reactions of Victorian nurses to the national review of specialist nurse education. AB - This article reports on the opinions of specialist nurses practising in Victoria toward recommendations proposed in the National Review of Specialist Nurse Education (Specialist Review) (Russell et al 1997). Seventy-five nurses undertaking a range of specialist nursing courses offered by three educational providers (a university, major teaching hospital and professional organisation) completed a survey questionnaire to establish the extent to which they agreed with the major recommendations from the Specialist Review. Results showed strong endorsement for most recommendations but with less agreement about issues of minimum credentials and future provisions. Comparison of responses across age, experience, employment and professional education subgroups identified significant differences of opinions on some recommendations. Findings from this study are discussed in relation to the Specialist Review and to implications for the future development of specialist nursing education in Australia. PMID- 10846987 TI - Programmatic research. A desirable (or despotic?) nursing strategy for the future. AB - Programmatic research is a planned and purposeful strategy in the development of a nursing discipline. We discuss the case made for programmatic research by international scholars as a determinant of scholarship and professional advancement, as well as issues about whether it should be knowledge or methods driven. As an example, the development of a clinical program involving the establishment of 'nursing beds' in the United Kingdom is described, together with a published critique of the program. While the literature portrays an overwhelmingly positive outlook for programmatic research, there are some cautionary tales to be told. We address these by way of the literature and personal experience--especially relating to tensions created by fixed research agendas, inability of researchers to follow their own research interests and funding difficulties. Potential disadvantages of programmatic research are also included. We conclude with suggestions as to how research students, early career researchers, and experienced researchers might become involved with programmatic research in positive ways. PMID- 10846988 TI - Residential aged care--accreditation and infection control. AB - As we approach the new millennium, many changes are becoming evident in all areas of health care. In particular, the accreditation of residential aged care facilities (RAC), already begun, is becoming a major agent of change both for the way in which RAC facilities operate as well as being a requirement for continuing Commonwealth funding. PMID- 10846989 TI - Consent to treatment and refusal of treatment by minors. PMID- 10846990 TI - Goods and services tax. PMID- 10846991 TI - The origin of the Roman Christian hospital. PMID- 10846992 TI - Ethics in practice. PMID- 10846993 TI - Establishing a cardiac surgery recovery unit within the post anaesthesia care unit. AB - At the Austin & Repatriation Medical Centre (A&RMC) an innovative approach was taken to improve the reliability of ICU bed availability for elective cardiac surgery patients. A cardiac surgery recovery unit (CSRU), a dedicated three bed area, was established within the Post Anaesthesia Care Unit (PACU) so as to deliver care to a group of patients that had previously been provided in the Intensive Care Unit (ICU). Derived benefits arising from this initiative include a reduction in the number of cardiac surgery cancellations for want of ICU beds, reduced length of stay of patients in the CSRU and in the Medical Centre, and an improvement in the level of knowledge and skills of PACU nurses in relation to caring for the specific needs of acutely ill patients. PMID- 10846995 TI - Ethics in clinical nursing supervision--an introduction to the theory and practice of different supervision models. AB - In clinical nursing supervision there are many complex ethical problems. This paper gives an answer to the question of what different clinical supervision models are available, explored from an ethical point of view. Four tentative ethically oriented supervision models, more or less related to the clinical supervision models, were found: decision-making oriented, case-oriented, principle-oriented and value-oriented. Questions of ethical issues and dilemmas related to the clinical supervision process have been addressed specifically to nurse clinical supervisors. It is interesting to note that the three dilemmas found in this study are related to the relationship between supervisor and supervisee. Clinical nursing supervision is an ongoing systematic process that encourages and supports both improved professional and improved administrative practice. In this process the nurse clinical supervisor is committed in her/his role to compromise and to take responsibility for the supervisee's professional development. PMID- 10846994 TI - Evaluation of a model of collaboration: academic and clinician partnership in the development and implementation of undergraduate teaching. AB - The difficulty nursing students experience in making the transition from the university to clinical context is attributed to the gap between theory and practice, and education and service. Collaboration between academics and clinicians in the provision of undergraduate education is considered to be a strategy for overcoming these problems. A project team consisting of four academics and six clinicians collaboratively developed and implemented an acute care topic, in the third year of a pre-registration nursing course at the School of Nursing, Flinders University. A review of the literature did not provide examples of collaborative models relevant to undergraduate teaching. The subsequent model, devised by the project team, focused on technical, cultural and interpersonal aspects of collaborative teaching. The model was evaluated by illuminative research methods. Participants in the project included academics (n = 4), clinicians (n = 6) and students (n = 104). Student questionnaires and academic and clinician interviews were used to inform evaluation procedures. The qualitative data was coded and analysed based upon the definitions and characteristics of collaboration described by Henneman et al (1995). Study findings demonstrated that the model was effective in facilitating collaborative relationships necessary for the successful development and implementation of reality-based learning for students. PMID- 10846996 TI - Does the nursing diagnosis form the basis for patient care? AB - There is a growing level of activity supporting the development of a taxonomy for nursing diagnoses, interventions, and outcomes. While this development is occurring at a theoretical level, more work is necessary at a clinical level in order to determine how these concepts are being operationalized; more specifically, to establish whether there is a link between assessment, diagnosis and interventions documented in the patient's medical record. Using a retrospective audit this research reviewed 304 patient records. The study data revealed that the nursing diagnoses documented were not always supported by patient assessment information. It also illustrated inconsistent links between the documented nursing diagnosis and interventions. These findings challenge the a priori support for nursing diagnosis forming the basis for patient care, as it exposes clinical application weaknesses. Given the International Council of Nurses move to develop and evaluate an International Classification of Nursing Practice, these weaknesses need to be considered in any evaluation. PMID- 10846997 TI - Muriel Knox Doherty--her story. PMID- 10846998 TI - Influenza. AB - In Australia in April this year, there were 25 cases of influenza A and 8 cases of influenza B reported. Most of these, 11 of type A and all 8 type B occurred in Western Australia. In May, 33 type A and two type B were reported. Of these, 25 type A were in South Australia and the two type B were in Victoria. In addition, two type H3N2, an epidemic strain, were reported in Victoria. Most recently, in June 1999 36 type A (14 in Victoria and 12 in Western Australia) plus one case of H1N1 9 (also an epidemic strain) was reported in Victoria and 10 type B (seven in Western Australia and three in Victoria) were reported. This brings the numbers for the year to 263 type A and 48 type B. Although it was anticipated that this year could see the arrival of an influenza epidemic in Australia, this has not yet occurred. PMID- 10846999 TI - World Wide Web: mental health. PMID- 10847000 TI - Lighten the load. PMID- 10847001 TI - Roman military hospitals. PMID- 10847002 TI - Celebrations and challenges. PMID- 10847003 TI - Conference round up. PMID- 10847004 TI - The Paddington rail disaster. PMID- 10847005 TI - Identification and management of frostbite injuries. PMID- 10847006 TI - Acid and alkali injury. PMID- 10847007 TI - Comparing the use of ketamine and midazolam in emergency settings. PMID- 10847008 TI - Photographs and memories. PMID- 10847009 TI - We've caught up in more ways than one. PMID- 10847010 TI - Emergency care in Kosovo. PMID- 10847011 TI - Antipsychotic drug overdose. PMID- 10847012 TI - Photographs and mementos. The emergency nurse's role following sudden infant death. PMID- 10847014 TI - No trolley waits at all. PMID- 10847013 TI - Nurse practitioner autonomy in a clinical setting. AB - The ENPs who participated in this study were providing services to a group of patients similar in age and gender to those using emergency services at a national level. The ENPs were employed in a variety of settings and provided health care to patients with non-urgent through emergent conditions. They ordered less diagnostic tests for patients than providers in a national study, performed various invasive and therapeutic procedures as well as diagnostic procedures, and referred all patients except those transferred to the morgue or who left against medical advice for follow-up care. The ENPs had a slightly higher percentage (94 per cent vs. 84.4 per cent) of patients discharged home than providers in a national study. PMID- 10847015 TI - Telemedicine--the technology and its applications. PMID- 10847016 TI - The therapy of touch. PMID- 10847017 TI - Plant poisoning. PMID- 10847018 TI - The emerging role of the nurse practitioner in A&E. PMID- 10847019 TI - Role modelling as a teaching method. PMID- 10847020 TI - Maltreatment of children. AB - Child maltreatment is comprised of four major categories. These include physical abuse, physical neglect, sexual abuse, and emotional abuse. Theory development has evolved to explanatory models. Research has provided a great deal of insight into the recognition of child maltreatment, the factors that place children at risk for maltreatment, and the factors that place a caregiver at risk for becoming abusive. The emotional effects of child maltreatment on the child include shame, aggression, delinquency, criminology, depression, and symptoms consistent with posttraumatic stress disorder. Theoretical frameworks for child maltreatment intervention both with the child and the perpetrator have been developed. Theory testing and development should continue to provide direction for further intervention research. PMID- 10847021 TI - A comparative study of health, developmental, and behavioral factors in preschool children of battered and nonbattered women. PMID- 10847022 TI - The health care provider's role in the disclosure of sexual abuse: the medical interview as the gateway to disclosure. PMID- 10847023 TI - Integrating a neurobiological systems approach into child neglect and abuse theory and practice. PMID- 10847024 TI - Identification and treatment of child physical abuse through medical and mental collaborations. PMID- 10847025 TI - Rethinking relationships between divorced mothers and their children. PMID- 10847026 TI - Review, critique, and guidelines for the use of herbs and homeopathy. PMID- 10847027 TI - Surfactant lavage for meconium aspiration syndrome: a pilot study. PMID- 10847028 TI - Children's responses to immunizations: lullabies as distraction. PMID- 10847029 TI - A survey of skin care practices for premature low-birth-weight infants. PMID- 10847030 TI - New developments in the treatment of HIV disease: an overview. PMID- 10847032 TI - To be thin is in. Or is it? Recognizing and measuring adolescent eating disorders. PMID- 10847031 TI - Medicaid and managed care changes. Has care for children really improved? PMID- 10847034 TI - Report of the ASHA National Injury and Violence Prevention Task Force. An executive summary PMID- 10847033 TI - Emergency Medical Services for Children. PMID- 10847036 TI - Improve quality of staff, equipment, patient flow. PMID- 10847035 TI - The ophthalmology patient's pain. PMID- 10847038 TI - Echography (ultrasound) procedures for the Collaborative Ocular Melanoma Study (COMS), Report no. 12, Part I. PMID- 10847037 TI - Can pupillometers prevent potential problems? PMID- 10847039 TI - Digital imaging: new views of retinal pathologies. PMID- 10847040 TI - Diagnostic test & instrumentation. PMID- 10847041 TI - Creating stereograms for near point of convergence exercises. AB - Pencil push-ups, for near point of convergence exercises, are a basic treatment for symptoms related to near work. Utilizing the computer to generate training stereograms increases the patient's interest and compliance with NPC exercises and assures a better treatment result. PMID- 10847043 TI - Contact lenses meet cosmetic needs of patients. PMID- 10847042 TI - Treatment of epibulbar limbal dermoids. PMID- 10847044 TI - Recycling in ophthalmology. PMID- 10847045 TI - How to 'surf' for Medicare billing and coding information: use Internet and print sources. PMID- 10847046 TI - New glaucoma technology looks beyond visual ability to the site of damage. PMID- 10847047 TI - Living with macular degeneration: creative strategies used by older women. PMID- 10847048 TI - Iris and pupil evaluation: key considerations. PMID- 10847049 TI - Echography (Ultrasound) Procedures for the Collaborative Ocular Melanoma Study (COMS), Report no. 12, Part II. PMID- 10847051 TI - Multifocal IOLs: delivering realistic expectations to cataract patients. PMID- 10847052 TI - ASCRS survey: reasons for IOL explantation vary by lens type. PMID- 10847050 TI - Contact lenses for the presbyopic patient. PMID- 10847053 TI - Case study: health behaviors of a young woman with multiple roles. PMID- 10847054 TI - Pain: clinical validation with postoperative heart surgery patients. AB - PROBLEM: To estimate the content validity of the nursing diagnosis of pain in postoperative heart surgery patients. METHODS: Observation of and interviews with postoperative heart surgery patients (N = 80), 40 experiencing pain and 40 without pain. FINDINGS: The group experiencing pain had statistically different results from the group without pain, with a higher frequency in the following 19 defining characteristics: verbal report of pain, discomfort, fear of reinjury, sleep disturbance, guarding behavior, distraction behavior, irritability, restlessness, facial mask of pain, increased heart rate, immobility, anxiety, loss of appetite, self-focus, withdrawal, impaired thought process, unusual posture, increased blood pressure, and changes in respiratory patterns. CONCLUSIONS: These defining characteristics support the idea that pain is a complex phenomenon of clinical interest to nursing that needs better understanding. Studies using the same defining characteristics in other sample groups of patients with acute and chronic pain might be useful in the refinement of this nursing diagnosis. PMID- 10847055 TI - Fecal impaction: a review. AB - TOPIC: Fecal impaction, a review of the literature. PURPOSE: To review the definition, diagnosis, causes, signs and symptoms, and treatment of fecal impaction. SOURCES: Published literature. CONCLUSIONS: While a great deal has been published on fecal impaction, there is limited research. Clarification of when constipation changes to impaction is ambiguous. Causes of fecal impaction are multiple, and people can be affected at all ages. Signs and symptoms of and problems associated with fecal impaction are varied. Research-based treatment of fecal impaction is very limited, and nonresearch based treatment varies. There is a need for additional research to clarify terminology, diagnosis, and interventions. PMID- 10847056 TI - Physical activity deficit: a proposed nursing diagnosis. AB - TOPIC: Physical inactivity is a major public health problem. Many of the leading causes of death and disability are associated with physical inactivity. The literature is filled with documentation about the importance of living physically active lifestyles. Healthcare professionals have started to incorporate health promotion into their practices, but there is still a large amount of work to do. PURPOSE: To discuss the label, definition, defining characteristics, and related factors of the proposed nursing diagnosis "physical activity deficit." This diagnosis has been submitted for consideration to the North American Nursing Diagnosis Association (NANDA). SOURCES: Existing empirical literature, professional journals, government documents, clinical handbook, text books, a conference statement, and a U.S. Surgeon General's report. CONCLUSIONS: Nurses are in an ideal position to lead when it comes to health promotion. Because physical inactivity is a modifiable risk factor for many diseases and disabilities, it is essential that it is included in nursing science. The diagnosis "physical activity deficit" would allow nurses to take a role in teaching the hazards of physical inactivity, explaining the benefits of activity, and encouraging regular exercise programs. PMID- 10847057 TI - Clarification of the NANDA/NDEC relationship. PMID- 10847058 TI - Keeping secrets no more. PMID- 10847059 TI - Clarifications, please. PMID- 10847060 TI - Flexibility: a concept analysis. AB - Flexibility is a quality that is deemed essential for nursing as the healthcare environment escalates into greater complexity. The word "flexibility" appears in nursing literature addressing the need to prepare for the next millennium, yet the concept of flexibility is rarely defined. Other disciplines, such as engineering, have struggled for years and continue to struggle to achieve conceptual clarity with regard to flexibility. A concept analysis using Rodgers' evolutionary method was undertaken to understand the meaning of flexibility. PMID- 10847061 TI - Videotaped focus groups: transforming a therapeutic strategy into a research tool. AB - Focus groups have become a popular method of collecting data for research projects. The addition of using a videotaped approach has broad implications for researchers, as it allows constant replay of the session. Although on the surface this approach seems straightforward and easy to use, the authors caution that this is not an approach for the novice. Investigators planning to use a focus group method must be well versed in group process. There is an unpredictable nature to the focus group process, and researchers need to prepare for the inevitable mishaps that lead to lost opportunities for data collection. PMID- 10847062 TI - Thirty-seven months as prisoners of war. 1945. PMID- 10847063 TI - Is nursing an academic discipline? PMID- 10847064 TI - My story: evolving a faculty-student practice in a homeless shelter. PMID- 10847065 TI - Changing a suprapubic catheter--2. PMID- 10847066 TI - The turnover of critical care nursing staff costs the NHS 5.5 m pounds a year. PMID- 10847067 TI - Critical to success. PMID- 10847068 TI - Mismatch of resources. PMID- 10847069 TI - Dead men's shoes. PMID- 10847070 TI - In the red. PMID- 10847071 TI - Let battle commence. PMID- 10847072 TI - If services are sensitive to users' needs, they are moving in the right direction. PMID- 10847073 TI - Hit and miss deal needs a rethink. PMID- 10847074 TI - The only way is up. PMID- 10847075 TI - Taken to task. PMID- 10847076 TI - A view to a skill. PMID- 10847077 TI - Self-regulation: apply within. PMID- 10847078 TI - Reaching Rabaraba. PMID- 10847079 TI - Backstage drama. PMID- 10847080 TI - Telephone banking. PMID- 10847081 TI - Web rings are an easy way of accessing relevant information quickly. PMID- 10847082 TI - The heart, Part Four: Basic cardiac arrhythmias. PMID- 10847083 TI - Pneumonia: still a deadly disease. PMID- 10847084 TI - Understanding cystic fibrosis, improving life expectancy. PMID- 10847085 TI - Not bad, just misunderstood. PMID- 10847086 TI - Bullets and bombs, cuts and crashes. PMID- 10847087 TI - Wound care. Concentrated care. PMID- 10847088 TI - Wound care. Cleansing rites and wrongs. PMID- 10847089 TI - [Evaluation by HIV/AIDS specialists of risk of HIV infection in university students, based on different sex practices]. AB - To detect the beliefs concerning the risk levels related to several sexual practices/habits and the HIV infection among undergraduate students and to compare their opinion with the AIDS experts were the objectives of this study. A 25-item questionnaire (Likert Probability Scale) about sexual practices/habits were answered by undergraduate students enrolled in the courses of nursing, medical school, pharmacy-biochemistry, psychology. Based on Factorial analysis and by using the Varimax Rotation System, 25 patients were distributed in 7 factors, and 5 patients were excluded. From the 20 patients, 5 were reunited in 2 factors and studied in this paper. Factor X reunited the patients 1 (vaginal intercourse with condom) and 2 (anal sex with condom). In factor Y the patients included were: 3 (heterosexual intercourse), 4 (vaginal intercourse without condom) and 5 (anal intercourse without condom). In 80% of the 5 patients, it was observed that the students have knowledge similar to the experts"s evidences But, continuing education is necessary to these people, as human beings and as future health care givers to the HIV infected/AIDS patients. PMID- 10847090 TI - [Evolution of the discipline "Administration Applied to Nursing" at the USP Nursing School 1980-1995]. AB - With the objective to describe the evolution of the discipline "Administration Applied to Nursing", in the period from 1980 to 1995, courses' plans texts write by the academic staff responsible for their elaboration and execution were studied. Texts of the items "Objectives and Content" were analyzed under the point of view of Phenomenology, being submitted to the process of analysis of speech defined by Martins and Bicudo. The result of the procedures used in the analysis, allowed the identification of six categories of thinking: Nursing Management, Ideological and Politic Bases for Nursing Management, Comunication as an Instrument to Managing, Management of Physics Resources, Management of Material Resources and Management of Human Resources. The modification in the courses' plans observed during the studied period, evidenced two important moments--one at the end of the eighties, were the reformulation of the ideological basis which the discipline basis was adapted and another at the end of the first five years of the nineties, with curricular reorganization. The identification of this course allowed the follow-up and comprehension of the development of the discipline, providing resources for analysis of the correlated disciplines of the same discipline in other contexts and experiences. PMID- 10847091 TI - [Special attention to women's health: significance and implications]. AB - The text is a reflexion about the political implications and practices related to the concept of integral attention of woman's health, in its several interpretations, for reference on the women multi-dimensionality. PMID- 10847092 TI - [Guiding patients' family members through an intensive care unit: difficulties of lack of systematization?]. AB - The purpose of this study was to analyse the problematic situation experienced by nurses at the moment of guiding the relatives of patients in an ICU. Therefore, seven (7) nurses who worked in this unit, which belonged to a general, public and training hospital in the city of Sao Paulo, were interviewed. The results have shown that the period of guiding brings anxiety and stress to the nurses, originated by the critical patient's condition, as well by the deficiency of conduts and in the systematization of this activity in the unit. Based on the results it was proposed the creation of na assistance plan. That includes the guiding to the family and the elaboration of a written instrument of orientation. PMID- 10847093 TI - [Research on child health: the life story method and mothers' oral statement]. AB - Children morbimortality rates in Brazil provide some indications about the life patterns of certain social groups, considered as risk groups, due to their vulnerability to a number of health impairments, but they do no display a clear picture of their quality of life. The present study aims to describe the steps undertaken by the authors to construct a qualitative research study on maternal living conditions and lifestyle, discussing certain aspects related to the study's design, such as, the definition of the research problem, the theoretical framework and data collection, particularly, the use of oral reports by mothers and the advantages and difficulties of this technique. Such narratives make it possible for researchers to come to know the needs, worries and strategies mothers use to care for their children's health, as well as, expose some wider social determinants of their life patterns. The knowledge thus generated is important to orientate the organization of policies and social practices to be developed by the health sector. PMID- 10847094 TI - [The right to information and the autonomy of the hospitalized elderly]. AB - It was carried out a study with the aim to analyse the autonomy of hospitalised elderly, based on the comprehension about their rights of information and the informed consent on proposals of diagnosis and therapeutics. The results showed patient's lack of information, dissatisfaction on the degree of information. It was also verified that the patient's family frequently acts as information intermediary between the health team and the patient. Therefore, autonomy process of decision making was compromised, as well as the informed consent of the elderly. PMID- 10847096 TI - [Living with cleft lip and palate patients: experience of a nurse]. AB - The purpose of this study was to understand the life of nurses in assisting the porter of cleft lip and palate In order to so we decided for a qualitative approach in the methodology of phenomenology. We interviewed nine subjects who took part of this study and their interviews were guided by this question: "How is it like to watch over/take care of patients with cleft lip and or palate?", The themes that emerged revealed the essence of the phenomenon in the nurse's perspective searching with the careful meanings to contribute to the improvement of assistance quality, thus offering subsidies to the development of a continuous educational program to the nursing team (staff). PMID- 10847095 TI - [Profile of HIV-positive women hospitalized in a public maternity ward in Rio de Janeiro]. AB - AIDS has been scaring the world population for the last decades. The writers of this article decided to investigate and research into the high incidente of women HIV+ hospitalized in a public maternity in the district of Rio de Janeiro, showing the profile of pregnant women from 1987 to 1996. After a thorough analysis of a total of 102 cases this number was eventually broken down showing that the majority of the female population are single, between the ages of 19 and 33 years, have low schooling, mainly housewives and were sexually contaminated. The results show that the affected groups lack the knowledge as to how they could protect themselves against contamination. It is, therefore, of vital importance to take the necessary measures to help prevent contamination and to focus on health education. PMID- 10847097 TI - [Suffering and pleasure experienced by nurses working at an intensive care unit of a university hospital]. AB - The purpose of this study is to grasp and to understand the nurses' social representations concerning their work in intensive care unit and the ways of expressing feelings of suffering and pleasure. Six nurses of the intensive care unit of medical, surgical and pediatric clinics were interviewed. The analysis of the social representations made possible the apprehension of the symbolic dimension of the work in the psycho-social (individual), social-dynamic (group) and institutional perspective. We verified that the work in intensive care unit brings pleasure to nurses despite of intense emotional constraint. This qualitative study allowed na in-depth reflection on the issue and made evident the need to deepen research. However, other investigations have to be carried out to deepen the subject on the subjective and symbolic dimension to subsidize the administration of human resources in nursing. PMID- 10847098 TI - [Bibliographic review of children's accidents]. AB - The study's proposal was to evidence a bibliography review about children' accidents. The bibliography and documental researches were used as methodology. The data made clear that children accidents are the bigger public health problem in England. In the United States, in 1989, had happened about 2,700 deaths as accident results in children under 14 years old. In Brazil has been registered high index of attendance at pediatric emergencies that involves home accidents. It has been concluded that these cases have been increasing and they need special attention and preventive approach. PMID- 10847099 TI - [Do we comfort? We deal with human beings without being aware of our humaneness]. AB - Reflections are intended to propose a confrontation between patients and families' thoughts, expectations, and needs regarding comfort and the usual behaviors adopted by health care agents during their interactions the clients. If comfort as viewed by patients is seen as a humanistic practice, which professional attitudes can become an obstacle to comfort? Reflections are also associated to factors that determine professional behaviors and the identification of the paths for a more humanistic practice. PMID- 10847101 TI - [Preliminary study of the relationship between the cardiopulmonary arrest time and its consequences in trauma patients]. AB - The proposal of this research was to obtain parameters to start or maintain cardiopulmonary resuscitation (CPR) in victims of trauma. The duration of the cardiac arrest and the CPR of the survivors was described, as well as the cerebral performance and the mortality of these victims 24, 48 and 72 hours after these events had happened. With the results of this characterization the relation between duration of cardiac arrest time, CPR and mortality were described. Data for this report were collected in Hospital das Clinicas da Faculdade de Medicina da Universidade de Sao Paulo emergency department. A big amount of the victims (93.4%) presents severe trauma and main cause of death was brain injury. Survival at 72 hours after CPR was 10%. The assessment, during the 72 hour period, of the survivors from cardiac arrest of traumatic cause has shown bad cerebral performance of those victims in that period of time. The survivor after the first episode of CPR was strongly related to cardiac arrest time when compared with CPR time. The time of cardiac arrest < or = 4 minutes and CPR < or = 20 minutes was related to survival more than 72 hours. PMID- 10847100 TI - [The patients' perception during their stay in the intensive care unit]. AB - The purpose of this study is to identify the patients's perception during their stay in the ICU. The sample was composed by ten patients, who had gone to cardiac surgery. They received psychological assistance only after their discharge from the ICU. The data was obtained throw a qualitative approach, using a content analysis. The results suggest that the patients have a stereotyped view about the ICU, linked with the idea of suffering and death; the nurses play an important role during fragility movements, physical and emotional dependence; the pain, by its subjective nature, individually and emotionally, is inevitable, because it is related to procedure and usually it is associated to physical suffering. PMID- 10847102 TI - [Making something more: a nursing student's experiences in nursing care within the family]. AB - This study sought to understand how nursing students experience nursing care in the family context. Phenomenology was used to interpret data collected using interviews with the students. Four themes were uncovered and MAKING SOMETHING MORE emerged as the phenomenon who integrated the motivational situation perceived by the students, their feelings, their actions, and their reflections about the lived experience. PMID- 10847103 TI - [Experiences of a mother of a Down syndrome child]. AB - The present thesis was developed based on "The Social Representations Theory" and its purpose was to understand the mother's representation of the Down syndrome child. The subjects were nine mothers of Down syndrome patients between the ages of six and twelve, at a Sao Paulo specialized facility. The study material was obtained through semi-structured and individual interview, and examined by means of content analysis, particularly the thematic analysis. The results pointed to a maternal representation of the child with a predominance of negative components. Based on that perception the mother experienced ambivalent feelings and behaved in a overprotective way. PMID- 10847104 TI - [Nature and severity of injuries in traffic accident victims]. AB - The objective of this retrospective study is to characterize the nature and severity of injuries of hospitalized traffic accident victims using the "Abbreviated Injury Scale" (AIS). Two-hundred and twenty such patients in a trauma reference hospital in Sao Paulo, Brazil were assessed. One-hundred and eleven of them were pedestrians, eighty-three vehicular passengers and twenty-six motorcyclists. The most common injuries were of the limbs, pelvic girdle and head/neck. Injury severity in all these patients was AIS = 3. Two-thirds of the forty-five victims who died were pedestrians. PMID- 10847105 TI - [Effect of pedagogic methods on planning of nursing education: bibliographic review]. AB - This paper looks historically and critically at the pedagogical and conceptual models using by nurses during five cycles of Brazilian History in nursing education. This article examines the patterns of nursing education while this historical time and suggests to include pedagogic models based on problem solving in nursing health practice. PMID- 10847106 TI - [The process of making laws]. AB - This paper presents a simplified summary of the process to make laws. It starts from the juridical concepts, requirements needed, hierarchy of the Brazilian laws, the need to follow proper steps through channels and committees within the National Congress, quorum required until its final approval with the presidential sanction, when a project of law becomes an enacted law. The purpose is to help nurses and other interested people to participate in the development of their profession through laws which better fulfill the professional interests of the group. PMID- 10847107 TI - [Requirements of academic standards in nursing schools: perspectives]. AB - The article presents perspectives to admission and development in the nursing teaching profession. PMID- 10847108 TI - [Attempts at innovations in the practice of teaching and practicing psychiatric nursing]. AB - In the context of recent changes that are taking place in the field of mental health care in Brazil, the University has the responsibility to redimensioning human resources qualification of mental health workers. At the Mental Health Care and Educational Program of the University of Sao Paulo and Sao Paulo Health Office (Sao Paulo/Brazil), an experience still in course, the nursing students attending to psychiatric nurse training evaluated that theoretical-practical education dealing with health care issue in the field of Psychosocial Rehabilitation purpose, at the same time that directly interferes in the qualification of practices, simultaneously develops the students attitudes and knowledge that qualify them to perform coherent practices in accordance with rehabilitation models of care. PMID- 10847109 TI - [Expectations of nursing students from their initial hospital training experiences]. AB - The first experience in hospital training may create tensions and anxieties. These feelings interfere negatively in the teaching-learning process. The aim of this paper was to investigate the expectations of Nursing Fundamentals II students regarding their initial hospital training. Second-year students of the Nursing Course offered by Universidade Estadual de Maringa (state of Parana, Brazil) were chosen as subjects of this investigation. The students revealed a state of anxiety when confronting technical procedures, relationship with patients and evolution. By reflecting on these aspects instructor the may increase the teaching-learning efficiency and humanize the professional training. PMID- 10847110 TI - [Technical levels at the hospital]. PMID- 10847111 TI - [Pharmaceutical satellite branches: for a more efficient supplying of care units. Interview by Eric Charles]. PMID- 10847112 TI - [Sterilization: a priority unit within the pharmacy service]. PMID- 10847113 TI - [The moveable hospital computer: medical record at the bedside]. PMID- 10847114 TI - Dofetilide for atrial fibrillation. PMID- 10847115 TI - Oral contraceptives. PMID- 10847116 TI - [Confidence intervals and their relevance for the interpretation of results. Audit of the journal "Strahlentherapie und Onkology"]. AB - BACKGROUND: The statistical quality of the contributions to "Strahlentherapie und Onkologie" is assessed, aiming for improvement of the journal and consequently its impact factor. MATERIAL AND METHODS: All 181 articles published during 1998 and 1999 in the categories "review", "original contribution", and "short communication" were analysed concerning the appropriate use of confidence intervals. RESULT: Forty-four publications were excluded from analysis, because they did not contain quantitative data or because the quotation of a confidence interval would not have been meaningful for other reasons. Of the remaining 137 publications only 27 presented all relevant results with clearly defined and correctly interpreted confidence intervals. This corresponds to a fraction of 20% (95% CI: 13-28%). CONCLUSION: Authors, peer reviewers, and editors could contribute to improve the quality of the journal by setting value on the documentation of confidence intervals. PMID- 10847117 TI - [Carbon ion irradiation of skull base tumors at GSI. First clinical results and future perspectives]. AB - BACKGROUND: Radiobiological and physical examinations suggest clinical advantages of heavy ion irradiation. We report the results of 23 women and 22 men (median age 48 years) with skull base tumors irradiated with carbon ion beams at the Gesellschaft fur Schwerionenforschung (GSI), Darmstadt, from December 1997 until September 1999. PATIENTS AND METHODS: The study included patients with chordomas (17), chondrosarcomas (10) and other skull base tumors (Table 1). It is the first time that the intensity-controlled rasterscan-technique and the application of positron-emission tomography (PET) for quality assurance was used. All patients had computed tomography for three-dimensional-treatment planning (Figure 1). Patients with chordomas and chondrosarcomas underwent fractionated carbon ion irradiation in 20 consecutive days (median total dose 60 GyE). Other histologies were treated with a carbon ion boost of 15 to 18 GyE delivered to the macroscopic tumor after fractionated stereotactic radiotherapy (median total dose 63 GyE). RESULTS: Mean follow-up was 9 months. Irradiation was well tolerated by all patients. Partial tumor remission was seen in 7 patients (15.5%) (Figure 2). One year local control rate was 94%. One patient (2.2%) deceased. No severe toxicity and no local recurrence within the treated volume were observed. CONCLUSION: Clinical effectiveness and technical feasibility of this therapy modality could clearly be demonstrated in our study. To evaluate the clinical relevance of the different beam modalities studies with larger patient numbers are necessary. To continue our project a new heavy ion accelerator exclusively for clinical use is planned to be constructed in Heidelberg. PMID- 10847118 TI - [First experiences with a noninvasive patient set-up system for radiotherapy of the prostate]. AB - PURPOSE: Highly conformal radiotherapy techniques require precise patient positioning. We report our first experience with a new cast system for fixation of the pelvis during stereotactically guided intensity modulated radiotherapy (IMRT) of the prostate with respect to positioning accuracy of the prostate. MATERIAL AND METHODS: The immobilization device consists of a custom-made wrap around body cast that extends from the abdomen to the thighs and a separate head mask, both made from Scotchcast, and attaches to a frame for extracranial stereotaxy. Sixteen CT-studies (> or = 25 slices, thickness: 3 mm) of 2 patients who were immobilized for IMRT of prostate tumors were evaluated with respect to set-up accuracy of bony structures and the prostate itself. CT-studies were performed immediately before or after a treatment fraction. Deviations of bony landmarks and anatomical landmarks inside the planning target volume were measured in all 3 dimensions. RESULTS: Mean patient movements of 0.15 +/- 0.3 mm (latero-lateral), 0.9 +/- 1 mm (anterior-posterior), 1 +/- 1 mm (tranversal vectorial error) and < 3 mm slice thickness (craniocaudal) were recorded using bony landmarks and 0.9 +/- 0.9 mm (latero-lateral), 1.8 +/- 1.5 mm (anterior posterior), 2.2 +/- 1.5 mm (transversal vectorial error) and < 3 mm (craniocaudal) using the confines of, or landmarks within the prostate. Standard deviations of absolute positioning error as an often used metric for positioning accuracy ranged between 0.3 and 1.7 mm in the transversal plane. The worst case transversal vectorial deviation for the prostate was 4.4 mm. Figure 4 summarizes the set-up accuracy of bony landmarks and the prostate. CONCLUSION: The presented combination of a body cast and head mask system in a rigid stereotactic body frame ensures reliable noninvasive patient fixation for fractionated extracranial stereotactic radiotherapy. It provides precise and reliable positioning of the prostate and meets the requirements for highly conformal radiotherapy such as IMRT. No further improvement of repositioning can be achieved with external immobilization devices since the positioning error of the target relative to the skeleton exceeds the accuracy of the positioning of the skeleton itself. PMID- 10847119 TI - Dose measurements in the build-up region for the photon beams from Clinac-1800 dual energy medical linear accelerator. AB - AIM: Since the skin dose becomes the limiting factor while deciding the tumorcidal dose, the detailed analysis of dose distribution in the build-up region is necessary for high-energy photon beams. In this study the beam characteristics affecting the build-up and skin dose for 6- and 18-MV photons are analyzed. MATERIALS AND METHODS: Measurements were made with 6- and 18-MV photons using a PTW parallel-plate ionization chamber (B 23344-036) and a RDM-1F electrometer. Build-up ionization measurements were made with the chamber fitted into a 25 x 25 x 25 cm polystyrene phantom with a fixed SSD of 100 cm. The entrance and build-up dose measurements were made with a polycarbonate and a mesh type metallic shielding tray and a 45 degrees wedge. Exit dose measurements were carried out for the graphite patient supporting assembly table top, 1.0 cm thick piece of wood and the 1.0 cm thick patient supporting perspex base frame for head and neck treatments. RESULTS: It was observed that the dmax decreased slightly with field size as with other accelerators. For both photon energies the surface dose was observed to increase with increase in field size. It was also noticed that the dose in the build-up region increases slightly when the polycarbonate secondary blocking tray is introduced with the increase in surface dose. The data show that the tray perturbation factor (TPF) at surface decreases steadily with tray-surface distance for both photon beams for all field sizes. It was noted that the TPF was more when the polycarbonate tray was introduced at shorter tray surface distances for both energies. At tray-surface distances above 60 cm the TPF almost remained close to unity for 6-MV photons for all field sizes, whereas the continuous decrease in TPF could be noted for 18-MV photon beams even after the TPF reached unity. CONCLUSION: The increase in surface dose with field size for both photon energies is due to the electron scattering from the intervening materials. The use of wedge filters absorbs low-energy scattered electrons significantly and hence, the relative surface dose (RSD) is always less than unity. The increase in dose enhancement percentage with graphite compared to perspex supporting assembly indicates that the electron backscatter is proportional to the atomic number of the medium. PMID- 10847120 TI - Chromosomal radiosensitivity, cancer predisposition and response to radiotherapy. AB - AIM: This paper briefly summarizes the research on this topic, undertaken in the Department of Cancer Genetics, Paterson Institute for Cancer Research, Manchester, England, over the previous 6 years. PATIENTS AND METHOD: Patients with the recessively-inherited disease, ataxia-telangiectasia (A-T), who are cancer-prone and suffer severe reactions after radiotherapy, also have highly radiosensitive cells, particularly when chromosome damage is used as the measure of radiosensitivity. Enhanced chromosomal radiosensitivity is also a feature of many other cancer-prone disorders. We have investigated the possible role of such radiosensitivity as a marker of cancer predisposition and response to radiotherapy in the general population. RESULTS: We found that 42% (57/135) of breast cancer patients exhibit chromosomal radiosensitivity when lymphocytes are irradiated in the G2 phase of the cell cycle, compared with 6% (6/105) of healthy controls (Figure 1). These figures are much higher than the estimated frequencies of carriers of the ataxia-telangiectasia gene (heterozygotes) amongst breast cancer patients (< 5%) and controls (0.5%). We have also obtained evidence of heritability of G2 sensitivity by studying relatives of breast cancer cases (Figures 2 and 3). The pattern of inheritance is relatively simple and attributable to 1 or 2 genes segregating in each family (Figure 4). In a prospective study of 123 breast cancer patients, 9 (7%) had severe acute reactions to radiotherapy and their mean G2 sensitivity was significantly greater (p = 0.001) than that of the remaining patients (Figure 5). In 16 patients with adverse acute reactions we found no mutations of the ataxia-telangiectasia gene (ATM). Using another chromosomal assay (micronucleus induction in G0 lymphocytes) we found that the mean radiosensitivity of patients with severe late reactions was higher than that of normal reactors. For example, 8 patients with severe fibrosis were more sensitive (p = 0.055) than 39 patients with a normal response (Figure 6). However, the discriminatory power of these chromosomal assays is too low for them to be used alone in a clinical setting. CONCLUSION: Our results provide good evidence that genes other than ATM, that confer chromosomal radiosensitivity, are involved in low penetrance predisposition to breast cancer in a high proportion of cases and contribute to adverse reactions after radiotherapy. PMID- 10847121 TI - [A simple method for transposing MRI information to simulation films. Technical note]. AB - BACKGROUND: The use of MRI information in the treatment planning process is still expensive and time consuming. Electro-optical devices or specially designed post processing software in here the risk of picture distortion. PATIENTS AND METHODS: Between June 1998 and June 1999 in 48 patients with brain tumors and 11 patients with carcinoma of the prostate CT treatment planning and MRI were performed with identical patient positioning using the same devices as in the simulation. The transposition of organ and tumor volumes between MRI and simulation film takes place by overlying both on the brightening screen by using a grid. MRI and simulation films must be produced using an identical magnification factor. RESULTS: For 9 patients with brain tumors and 2 patients with a carcinoma of the prostate the transposition of MRI information on the simulation films showed the necessity of a modification of the shielding or the treatment portals. CONCLUSIONS: The simple method of direct transposition of MRI structures into simulation films allows to accomplish a verification of the portals and conformation which result of the treatment planning process. Moreover an individualized shielding can also directly be realized. PMID- 10847122 TI - [Comparison of fluorouracil + folinic acid, fluorouracil + levamisole and fluorouracil + folinic acid + levamisole in Dukes' B and C colon cancer: NSABP Trial C-04]. PMID- 10847123 TI - [Plasma and salivary pharmacokinetics of 5-fluorouracil]. PMID- 10847124 TI - [Influence of psychological reactions to survival in breast cancer: a population based cohort study]. PMID- 10847125 TI - [Effect of treatment delay on survival in patients with breast cancer: a systematic review]. PMID- 10847126 TI - Hepatitis C viral quasispecies. AB - Analysing significant numbers of cDNA clones of the hepatitis C virus (HCV) from single isolates provides unquestionable proof that the viral genome cannot be defined by a single sequence, but rather by a population of variant sequences closely related to one another. This way of organizing the genetic information is referred to as quasispecies. Throughout HCV infection, the number and composition of the variants in the viral population keeps changing owing to environmental influences, resulting in a virus that is constantly redefining itself both genetically and phenotypically. Therefore, the virus has often been investigated in population terms. Many clinical studies have tried to unravel, through the parameters that characterize the HCV quasispecies, prognostic markers of the disease and its response to treatment. Other investigations have focused on discovering how the virus and host interact during chronic infection. The consensus sequence, the rate of fixation of mutations and the complexity of the viral population are useful parameters for describing the viral population behaviour and its interaction with the host. In addition to sequencing, several other methods, based on electrophoretic mobility, have been used to study these parameters, such as temperature gradient-gel electrophoresis, single-strand conformation polymorphism and gel-shift analysis. The viral region examined, the source of clinical specimen, as well as the methodology employed, will be decisive in interpreting the information obtained. PMID- 10847127 TI - Gene therapy of viral hepatitis and hepatocellular carcinoma. AB - Gene therapy represents an attractive approach to treat a great variety of diseases, both inherited and acquired, and it is moving slowly from a proof-of principle phase to a wide application in most medical fields. Liver cancer and viral hepatitis are natural targets for this new therapeutic alternative due to the lack of success of conventional antitumoral and antiviral treatments and the ominous prognosis related with liver tumours. Gene therapy for viral hepatitis is aimed to boost the patient immune response against viral antigens or to make cells resistant to infection by blocking the viral life cycle. Gene transfer techniques applied to the treatment of hepatocellular carcinoma include drug sensitization by suicide genes, genetic immunotherapy, normal tissue protection by transfer of the multidrug resistance gene, replacement of tumour suppressor genes, inhibition of oncogenes and modifications of the biology of the tumour (antiangiogenesis). However, major advances in our understanding of the regulation of gene expression, design of the expression cassettes and development of more efficient gene transfer vectors are mandatory before gene therapy can become a widely used therapeutic modality. PMID- 10847128 TI - Global surveillance and control of hepatitis C. Report of a WHO Consultation organized in collaboration with the Viral Hepatitis Prevention Board, Antwerp, Belgium. AB - Hepatitis C is a global health problem caused by infection with the hepatitis C virus. Although representative prevalence data are not available from many countries, available data indicate that approximately 3% of the world's population is infected with HCV. It is estimated that as many as 170 million persons world-wide may be infected with HCV. In many countries, the exact magnitude of the problem and the relative contribution of the various routes of transmission have not been defined with population-based studies. Wherever possible such studies should be performed to enable countries to estimate the burden of hepatitis C disease, to prioritize their preventative measures and to make the most appropriate use of available resources. To assess hepatitis C on a global scale, the World Health Organization (WHO) organized a consultation of international experts, in order to review the public health aspects related to hepatitis C infection and to make recommendations for its prevention and control. PMID- 10847129 TI - A prospective study of transfusion-transmitted virus transmission by blood transfusion. AB - Recently, a new single-stranded DNA virus (TT virus, TTV) has been isolated and related to post-transfusion hepatitis. The aim of this study was to investigate the prevalence of TTV in blood donors and blood recipients, and the incidence of TTV transmission by blood transfusion. TTV DNA and serum markers of hepatitis B virus (HBV) and hepatitis C virus (HCV), were examined in 130 blood recipients, and the presence of TTV was studied in their 340 corresponding blood donors. The prevalence of TTV infection was 10.6% (36/340) in donors and 8.5% (11/130) in blood recipients, before transfusion. Eighteen subjects (15.1%) were found to be TTV positive, after transfusion, in the 119 blood recipients without TTV before transfusion; at least one of the corresponding donors was TTV positive. There were 46 subjects with post-transfusion hepatitis virus infection, 45 with HCV infection (including seven co-infected with TTV) and two with HBV infection (including one co-infected with HCV and one co-infected with TTV). The recipient with TTV and HBV co-infection and three of the seven patients with TTV and HCV infection had alanine aminotransferase (ALT) levels higher than 90 Ul-1, but only two of the 10 isolated TTV infections had a mild ALT elevation. These results show that prevalence of TTV was high in blood donors and hospitalized patients, and isolated TTV infection is not related to significant ALT elevation. PMID- 10847130 TI - Prevalence of hepatitis viruses in an anti-human immunodeficiency virus-positive population from Argentina. A multicentre study. AB - The objectives of this study were to investigate the prevalence of infections with hepatotrophic viruses in an anti-human immunodeficiency virus (HIV)-positive population from Buenos Aires and to compare it among the main risk groups for HIV infection. Four hundred and eighty-four consecutive patients attending the HIV outpatients clinic were studied: 359 men and 125 women, median age 29 years (range 16-67 years); 35.5% had presented acquired immune deficiency syndrome (AIDS)-defining conditions. Two hundred and thirty-four patients were intravenous drug users (IVDU), 99 had homosexual and 142 heterosexual preference, seven had received blood transfusions and two had no risk factors. Hepatitis B surface antigen (HBsAg), and antibodies to hepatitis B core antigen (HBcAb) and to hepatitis C virus (anti-HCV) were investigated in all patients; antibodies to HBsAg (HBsAb) and IgG antibodies to hepatitis D virus (anti-HDV) in all HBcAb positive patients; hepatitis B e antigen and antibodies to HBeAg (HBeAg) in all HBsAg-positive patients; IgG antibodies to hepatitis A virus (anti-HAV) in the first 307 patients; and IgG antibodies to hepatitis E virus (anti-HEV) in the first 91 patients. As control groups, contemporary voluntary blood donors were studied for prevalence of HAV, HBV, HCV and HEV. The percentages of HBcAb, HBsAg, anti-HCV and anti-HEV (58.5, 14.5, 58.5 and 6.6%, respectively) were significantly higher in anti-HIV-positive patients than in control groups (3.2, 0.5, 1.0 and 1.8%, respectively) (P = 0.000). The prevalence of HBcAb was significantly higher in IVDU (72.6%) than in heterosexuals (33.8%) (P = 0.0001) and in homosexuals (59.6%) (P = 0.0189). The percentage of HBsAg was significantly higher in IVDU (19.2%) than in heterosexuals (6.3%) (P = 0.0004). Anti-HCV was significantly higher in IVDU (92.3%) than in homosexuals (14.1%) and in heterosexuals (33.1%) (P = 0.000 in both cases). The prevalence of anti-HDV was relatively low (1.9%). There was no difference in the percentage of anti-HAV between HIV-positive and negative subjects. In conclusion, there is a high prevalence of HBV and HCV infections in HIV-positive patients from our area. Drug use is the main route of transmission, but prevalence of HCV in patients with, probably, sexually acquired HIV infection is also higher than in the control group. The increased prevalence of HEV infection in HIV-positive individuals is another provocative finding that warrants further study. PMID- 10847131 TI - An outbreak of enterically transmitted non-A, non-E viral hepatitis. AB - Patients with isolated serum transaminase elevations of unknown cause are common in China. An outbreak of such disease took place in a technicians' school during 1996. To define the epidemic and determine the etiology, a study was carried out, which included investigation of epidemiological, clinical and histological features. The symptoms of this disease were mild. The major clinical feature was transaminase elevation, and all serum markers of known hepatitis viruses were negative. Although the course of disease in most patients was self-limiting, in a few it was prolonged and relapsed. Histological findings were mild portal hepatitis or non-specific reactive hepatitis. The disease first appeared in 1994, and this outbreak occurred after October 1996. A total of 381 people were affected and the prevalence was as high as 60.7%. Casual contact and small-scale food transmission were considered to be risk factors for infection and the epidemic was under control 2 months later following the introduction of preventive measures for gastroenteric infection. Viral genomic fragments from the so-called transfusion-transmitted virus (TTV) were detected in acute-phase sera and stool samples collected 2 weeks before onset. Therefore, this disease outbreak might be another form of enterically transmitted viral hepatitis, not related to hepatitis A and E. PMID- 10847132 TI - Serum levels of hepatitis C virus RNA predict non-response to interferon therapy: comparison of two commercial assays. AB - We compared two commercial assays for quantification of serum hepatitis C virus (HCV) RNA to investigate whether pretreatment levels of serum HCV RNA could predict the outcome of interferon (IFN) therapy. The Amplicor HCV Monitor test is based on a single, combined reverse transcription and amplification reaction carried out by the Tth DNA polymerase using specific primers for the 5' untranslated (UTR) region. The Quantiplex HCV RNA 2.0 assay is based on specific hybridization of viral RNA by synthetic oligonucleotides complementary to the 5' UTR and core regions of the genome, allowing equal quantification of the six major genotypes. Receiver-operating characteristic (ROC) analysis was employed to identify the best cut-off value (predicting patients who were non-responsive to treatment) with corresponding sensitivity and specificity values. Logistic regression analysis was performed using these cut-off values. We studied 133 consecutive patients with chronic hepatitis C enrolled in a prospecsustained responders was 5322 copies ml-1 by the Monitor test and less than 0.2 million equivalents ml-1 (MEq ml-1) by the Quantiplex assay; for the 115 non responders/relapsers, the median viraemia was 83,125 copies ml-1 and 1.128 MEq ml 1 for the Monitor test and Quantiplex assay, respectively. Spearman's rank test gave a correlation of 0.63 between assays. The best predicting cut-off values were 22,134 copies ml-1 for the Monitor test and 0.330 MEq ml-1 for the Quantiplex assay; their respective sensitivities and specificities were 72% and 75% for Monitor and 67% and 83% for Quantiplex. By logistic regression analysis, the age and gender-adjusted odds ratios of high vs low HCV RNA levels, defining the risk of non-response, were 10.6 (CI 3.1-35.7) for Monitor and 14.3 (CI 4.3 47.3) for Quantiplex. The two assays had comparable sensitivity for serum HCV RNA but they identified different predictive cut-offs for non-response to therapy. PMID- 10847133 TI - Failed adoptive immunity transfer: reactivation or reinfection? AB - A 26-year-old female bone marrow transplant (BMT) recipient was hepatitis B surface antigen (HBsAg) and hepatitis B e antibody (HBeAb) positive. The donor, her human leucocyte antigen (HLA)-compatible sister, was HBsAg negative but hepatitis B surface antibody (HBsAb) and hepatitis B core antibody (HBcAb) positive. Twelve weeks post-BMT the patient became HBsAg negative, as determined using a monoclonal antibody-based assay. At 16 weeks post-BMT, HBsAg became undetectable by monoclonal and polyclonal immunoassay with seroconversion to HBsAb; however, at 24 weeks post-BMT the patient again became HBsAg positive. Both the recipient and the donor were retrospectively tested by hepatitis B virus (HBV) polymerase chain reaction (PCR) and found to be positive. The recipient displayed variants at amino acids 4 and 47 of the surface (S) gene prior to BMT. These mutations were not detected 32 weeks post-BMT when the S gene sequence was identical to that of an adr prototype. The donor was found to have four unique amino acid substitutions at positions 30, 98, 101 and 210 of the S gene. However, in vitro-expressed HBsAg from the donor was detected by commercial kits and an immunofluorescence assay, indicating that antigenic alteration did not explain HBsAg negativity. This donor highlights the value of PCR as the gold standard test for current HBV infection. It also demonstrates that discordance between two commercial HBsAg assays may not always be caused by antigenic variants. The second episode of hepatitis may theoretically have been caused by reactivation, selection of an escape mutant by HBsAb, reinfection or recombination. We suggest it was reactivation because none of the donor variants was seen in the recipient post-BMT. PMID- 10847134 TI - [The proarrhythmogenic activity of non-anti-arrhythmia drugs. Is treatment with antihistamines and cisapride safe?]. AB - The non-antiarrhythmic drugs, which possess antiarrhythmic properties could induce dangerous, potentially fatal arrhythmias--extrasystoles, ventricular tachycardia, sudden cardiac arrest. The arrhythmogenic properties are due to block of the potassium channels of the cells and are realized by prolongation of the QT interval on ECG. Accelerating mechanisms are the bradycardia and the hypokalemia. Such drugs are the H1 blockers--astemisol (hismanal) and terfenadine, the prokinetic cisaprid (prepulsid, propulsid, coordinax) and the macrolides. These preparations should be carefully prescribed and not combined with each other, as well as, with antiarrhythmics and blockers of the cytochrome oxidase system (antifungal antibiotics, metronidazole, cyprofloxacin, antidepressants). During their use the patients have to be followed up for changes in QT, bradycardia, arrhythmia, hypokalemia. PMID- 10847135 TI - [The extrahepatic manifestations in hepatitis C virus (HCV) infection]. AB - The hepatitis C virus (HCV) infection could affect not only the liver, but also other tissues, organs and systems. The number of the reported in the literature extrahepatic lesions by HCV incessantly increases. A reliable association between the infections by HCV and the mixed cryoglobulinaemias, membrano-proliferative glomerulonephritis and porphyria cutanea tarda is confirmed. The participation of HCV in the pathogenesis of some diseases of the thyroid gland, the lymphocytic sialadenitis, lichen planus, diabetes mellitus, thrombocytopenia, antiphospholipid syndrome, etc., is assumed. The extrahepatic lesions by HCV are probably connected with the participation of the immune system, but they may be as well due to the replicating virus in the affected tissues, organs and systems. The pathogenetic mechanisms of the extrahepatic and autoimmune manifestations of the infection with HCV are not elucidated, which poses difficult therapeutic problems regarding the choice of interferon and/or corticosteroid hormones. PMID- 10847136 TI - [Successful prophylaxis in a pregnancy with thrombophilic states]. AB - It has been observed that the presence of antiphospholipid antibodies (aPL) is associated with a high incidence of fetal loss and this is thought to--a result from the vasculitis of placental circulation and subsequent thrombosis. A 38-year old woman with second fetal loss had received oral anticoagulant after pulmonary embolism. APL were examined--anticardiolipin antibodies (aCA) were found. The third pregnancy was established and sintrom was stopped and replaced by fraxiparine 0.3 ml/24 h up to the end of the pregnancy even 20 days after delivery of a healthy baby. PMID- 10847137 TI - [The treatment of reflux esophagitis in patients with a Helicobacter infection of the gastric mucosa]. AB - Among the pathogenetic mechanisms for the occurrence of gastroesophageal reflux disease, Helicobacter pylori infection is indicated. The conclusions are absolutely opposite. The object of our work was to assume ex juvantibus to what extent the eradication of the Helicobacter pylori infection would accelerate the healing of patients with reflux oesophagitis and would reduce the number of relapses. To investigation were submitted 42 patients with reflux oesophagitis with confirmed Helicobacter pylori infection, classified according to Savary Miller. A group of 22 patients was treated 10 days with triple drug combination of omeprazole, amoxillin and metronidazol with the purpose of eradication of the infection, after which they continued with ranitidin up to 30 days, and a second group of 20 patients treated only with ranitidine for 30 days. The subjective complaints, endoscopic finding and present infection were followed up. A considerably higher number of recovered patients after eradication of the Helicobacter infection was established and the number of relapses for the six month period of observation was considerably reduced. PMID- 10847139 TI - [Deafness as an autoimmune phenomenon in a female patient with ulcerative colitis and autoimmune hepatitis]. AB - In the last years it was confirmed that deafness could occur as an autoimmune phenomenon in patients with ulcerative colitis. The object of the work is to demonstrate by means of retrospective analysis the occurrence of deafness in a female patient with ulcerative colitis, appearing in early childhood with underestimated clinical signs and confirmed after clinical manifestation of autoimmune hepatitis. The notion of deafness as an autoimmune phenomenon in ulcerous colitis could allow its timely treatment, restoration and preservation of hearing. PMID- 10847138 TI - [The prevention of ulcer relapses in patients with an ulcerative anamnesis in clinical and endoscopic remission following the eradication of Helicobacter infection of the gastric mucosa]. AB - The contemporary scientific consensus recommends treatment of the Helicobacter pylori infection in patients with history of peptic ulcer independently of the stage of activity. The object of the study was to confirm the necessity for treatment of Helicobacter pylori infection in patients with duodenal peptic ulcer in stage of clinical and endoscopic remission. To investigation were submitted 55 patients divided into in two groups--30 patients treated with triple medicinal combination of metronidasol 2 x 20 mg, amoxycillin 2 x 1000 mg and metronidasol 3 x 500 mg for 7 days. The second group of 25 patients was without medicinal therapy. The observation continued for 12 months. The patients with eradicated Helicobacter pylori infection underwent significantly less relapses, while the quality of their life considerably improved. PMID- 10847140 TI - [In Process Citation] AB - The monograph embraces high number of Bulgarian, Russian, English, American, German, etc. literature sources by most distinguished surgeons--scientists and lecturers, as well as proper long- year operative, scientific and tuition experience. The author treats 93 topics of the abdominal surgery. They are presented in accessible manner with emphasis laid on the most essential, necessary for the favourable outcome of the operation. Therapeutic results are reported, comparable to the best, published by internationally renowned authors. Many single and group illustrations--drawings and photos--are applied. The problems of abdominal surgery are discussed in accessible manner for physicians (specialisilising surgery, internal diseases, anaesthesiology, urology and orthopaedics) and scientific workers, as well as for medical nurses, rehabilitators and kinesitherapeutists. The trend emerges for organ-sparing reconstructive operations, etio-pathogenetically expedient, with biologic adaption of the patients, but also extended blockoperations and simultaneous operations in synchronic diseases. PMID- 10847141 TI - [The Index Pharmacorum Bulgaricus and its database--an automated drug information system]. AB - Authors represent an unique in its structure and comprehensiveness drug information system (DIS) produced by a team of professionals in different branches of science, taking them many years of creative efforts. The basic principles on which the information has been processed and classified, as well as the respective advantages and good possibilities for the usage of this information product have been explained. The printed version of DIS is represented, too. It is the 9-volume drug guide Index Pharmacorum Bulgaricus (IPB). This reference book has found a very good acceptance among the authorities in different fields of the medicine due to the completeness, accuracy and actuality of the pharmacological information included. PMID- 10847142 TI - [Tuberculosis and mycobacterioses--current problems]. AB - Some quantitative and qualitative changes, occurring in the tuberculous endemic during the last years are being analyzed. The analysis of the indices for the dynamics of the tuberculous endemic during the last years in our country reveals that the control of tuberculosis should take in consideration the following facts: 1) the increase of the morbidity will continue; 2) the increase of the morbidity disease is on behalf of the pulmonary tuberculosis. The causes in our country, which play role in the increase of the morbidity, are clarified. To detailed consideration are submitted the diagnostic problems, which are posed by the mycobacterioses in general and by the mycobacterioses and tuberculosis in HIV infection and AIDS. PMID- 10847143 TI - [Latex allergy]. AB - An avalanche-like increase of the cases with LA during the last decades is being noticed. The pathogenesis of LA is considered, the main latex allergens are characterized and their sources are presented in details. The risk factors and risk groups for the occurrence of LA are described, as well as the clinical presentation of the disease in its different expressions. The diagnostic methods for confirmation of LA are enumerated and evaluated. The significance of prevention is emphasized. PMID- 10847144 TI - [Secondary systemic amyloidosis A combined with primary glomerulonephritis and systemic diseases]. AB - Amyloidosis is characterized by organic dysfunction as a result of deposition of amyloid substance in the walls of the small blood vessels and extracellularly in different organs. The involvement of the kidneys in systemic amyloidoses AL and AA has irreversible evolution to renal failure. The object of the study was to determine the prevalence of the secondary (reactive) systemic amyloidosis AA in combination with primary glomerulonephritis (PGN) and lupus nephropathy (SLE) and to create diagnostic approach for its early detection. The prevalence of amyloidosis among the renal biopsies in the Department of Nephrology by the Chair of Internal Diseases for the period 1981-1988, retrospectively, is 4% (in 11 out of 268 biopsies). For the period 1989-1996, prospectively, by directed quest, amyloid was found in 35 out of 269 renal biopsies (11%). For differentiation of AA preliminary treatment of the histologic material with KMnO4 was used. In 20 cases amyloidosis appeared as independent finding in the renal tissue, while in 15 it was combined with histologic picture of immune nephropathies: in 11 with primary glomerulonephritis (7% out of 155 PGN) and in 4 with systemic lupus erythematodes (11% out of 31 SLE). The combination of PGN with AA was almost always associated with chronic infections. It was most often observed in diffuse membranous GN or FSGSH. Our studies demonstrate increased prevalence of amyloidosis among the renal biopsies during the last years, which could be due to directed quest, but it could be a real fact, too. We suggest staining for amyloid in all renal biopsies, as well as its directed quest in mucosae of the gastro intestinal tract and by aspiration of the abdominal subcutaneous fatty tissue in the patients with primary GN or systemic diseases. PMID- 10847145 TI - [The systemic and local immune responses in patients with alcoholic liver cirrhosis depending on hepatitis C viral infection (HCV)]. AB - The systemic and local immune response was studied in patients with alcoholic liver cirrhosis and the significance of the combined infection with HCV. To investigation were submitted 23 patients (16 males and 7 females) aged between 29 and 61 years with alcoholic liver cirrhosis. Of them 14 were anti-HCV(+) and 9 anti-HCV(-). As controls were used 36 clinically healthy individuals, matched by sex and age to the patients. The flow cytometric analysis of the lymphocyte (Ly) populations from the peripheral venous blood and of cells from liver aspirate obtained by blind liver biopsy according to Menghini, was performed with FacsTAR (Becton Dickinson). In the anti-HCV(-) patients, as compared to the controls (patients/controls) the Ly subpopulations were increased: CD3+/mm3:2010 +/- 738/1440 +/- 388; CD4+/mm3:1350 +/- 441/991 +/- 442; IL-2R+/mm3:133 +/- 78.5/31 +/- 20. In the anti-HVC(+) patients we established increased IL-2R+/mm3: 170 +/- 126 as compared with the controls and anti-HCV(-) patients. The suppressor/cytotoxic (CD8+) Ly with their suppressor (CD8+CD11b+) and cytotoxic (CD8+CD11b-) subpopulations and natural killers (CD16+) had a tendency to diminution in the anti-HCV(+) patients. In both examined groups the B (CD19+) Ly were non-significantly increased. The flow cytometric analysis of the cells from the liver specimen in 9 patients of whom 3 anti-HCV(-) and 6 anti-HCV(+) revealed that CD3+ on the average were 32.8% +/- 20.4% (from 9.2% to 65.1%); CD4+ were 21.1% +/- 7.4% (from 12.0% to 34.5%); CD8+ 22.6% +/- 11.8% (from 4.7% to 39.8%) and their values were higher in the anti-HCV(+) patients; the correlation CD4+/CD8+ = 1/1.09 +/- 0.6; CD16+ were 12.9% +/- 10.1% (from 1.9% to 34.8%); CD19+ varied from 3.2% to 27.8%; monocytes (CD14+) were 7.69% +/- 5.65 (from 2.0% to 15.8%) from the cells of the aspirate and their percentage contents was higher in the anti-HCV(+) patients. The results of out study revealed that in patients with alcoholic liver cirrhosis changes in the cell immune response were also observed and that they were more marked in infection with HCV. PMID- 10847146 TI - [Arterial hypertension and obesity--a dangerous combination]. AB - The combination of obesity with arterial hypertension is frequent finding in clinical practice. In 70% of the males and 61% of the females the high blood pressure is directly connected with obesity. The assumed mechanisms by which obesity leads to arterial hypertension are: insuline resistance; genetic factors (hypothesis for the sparing gene); correlations leptin-neuropeptide Y; fatty tissue as origin of local pressor and depressor humoral factors. The arterial hypertension in obesity is salt-sensible, associated with increased intraglomerular pressure, microalbuminuria and increased risk for cardiovascular complications. The reduction of the body weight is the principal nonmedical mean for treatment of the arterial hypertension. Of the antihypertensive drugs those which are neutral with respect to the carbohydrat and fat metabolism are preferred inhibitors of the converting enzyme, calcium antagonists, selective alpha-1 blockers, central alpha-2 agonist. PMID- 10847147 TI - [Cellular immune deficiency in patients with chronic glomerulonephritis]. AB - In our previous work we found quantitative changes in peripheral blood lymphocyte subpopulations (decrease in total T lymphocytes, increase in T suppressor cells and monocytes) in patients with idiopathic chronic glomerulonephritis (CGN). The aim of this study was the functional state of the immune response (lymphocyte proliferation and cytokine secretion in vitro) in order to characterize the cellular immune defects and their changes under immunomodulatory therapy. We studied 34 patients with active CGN, divided in groups treated by IVIG or combined corticosteroid and immunosuppressive therapy. We found decreased proliferative ability of PBMNC to mitogen as well as to antigen. There were increased basal production of TNF and sIL-2R and lack of increase of LPS-induced IL-1 in vitro. These deviations suggest a deficiency in cellular immune response in patients with chronic GN which was influenced by immunomodulatory therapy alongside with beneficial clinical effect. PMID- 10847148 TI - [Pain. Headache. The therapeutic possibilities]. AB - Pain is a disagreeable subjective sensation. It is an emotional experience, related to actual or potential damage of the tissues and is a phenomenon, which is perceived unconsciously. As a frequently encountered suffering, it has definite social and economic significance. According to the pathogenesis, the pain also could be somatogenic, neurogenic and psychogenic. It has different site of occurrence: somatic (internal or external) and visceral. It evolves acutely or chronically. Clinically it is expressed as headache--primary (migraine or of straining type) and secondary (symptom of a definite illness). The cause of the pain could be inflammation (of the teeth, back pain, distortion, etc.), elevated temperature, etc. The convulsive pains could be related to menstruation. The modern treatment of the pain is carried out with nonsteroid antiinflammatory agents, which possess marked to different degree analgetic, temperature lowering and antiinflammatory action (aspirin, paracetamol, ibuprofen, etc.). In pain and particularly in headache the preparation Tomapyrin tabl. (Boehringer Ingelheim) is effective, which contains 250 mg acetylsalicylic acid, 200 mg paracetamol and 50 mg coffein. The alkaloid coffein stimulates the CNS, strengthens the striated musculature and displays marked vasoactive effect. The redistribution of the blood mass explains its effect in migraine. For exerting influence upon the pain relaxation programs have been created, physical therapy and training for mastering the stress are carried out. PMID- 10847149 TI - The water quality of UK rivers entering the North Sea. PMID- 10847150 TI - River water quality in the Humber catchment: an introduction using GIS-based mapping and analysis AB - The regional water quality of the Humber catchment was mapped for key inorganic chemical determinands using a GIS system and an extensive Environment Agency and LOIS monitoring database. The resultant maps revealed the major factors affecting the general characteristics of regional water quality. Sewage inputs from industrial and domestic sources account for the high concentration of many determinands in urban areas. The concentrations of particulate components increase in tidal zones because of sediment trapping and tidal re-suspension effects. Some determinands also exhibit localized high concentrations related to coal mine drainage, soil pollution caused by past ore mining, bedrock geology, the agricultural use of fertilizers and the ingression of seawater into the estuary. PMID- 10847151 TI - Pollution regimes and variability in river water quality across the Humber catchment: interrogation and mapping of an extensive and highly heterogeneous spatial dataset AB - Water quality inter-relationships based on an extensive Environment Agency database are used to identify seven key types of clean and polluted riverine environments within the Humber catchment. These types were selected on the basis of linear separations between average concentrations of ammonium, chloride, dissolved nickel and chromium and acid-available particulate nickel and chromium. However, a more detailed analysis of determinand and flow relationships at each type locality revealed complex patterns due to the highly variable nature of pollutant sources at the local scale. The information presented contrasts systems at a larger scale, where simpler integrated features are observed, and which can be represented by two-component mixing models. Scale and hydrology seem to be important features in determining the simplicity or complexity of determinand inter-relationships, and the implications of these findings are discussed. PMID- 10847152 TI - A new approach to deriving 'best-estimate' chemical fluxes for rivers draining the LOIS study area AB - A central objective of the LOIS Community Research Programme was to provide estimates of the land-ocean flux of chemical species from the LOIS study area. In the absence of detailed information on the concentration of most of the dissolved and particulate-associated chemical constituents, it has proved necessary to make use of the relatively infrequent samples collected in the UK Harmonised Monitoring Scheme, in order to estimate chemical fluxes for the LOIS rivers. In these circumstances, standard procedures may provide unreliable results, and a new approach has been developed to derive improved estimates of chemical flux from the LOIS study area. This approach is based on the generation of detailed synthetic time series of concentration, which are used to estimate the likely errors associated with different flux calculation procedures, especially those resulting from differences in the timing of chemographs and hydrographs during storm events. The approach was applied on a river by river and determinand by determinand basis to identify and to employ the best methods for computing fluxes of 45 chemical determinands from the LOIS study area during the period 1992-1996. PMID- 10847153 TI - Riverine inputs of major ions and trace elements to the tidal reaches of the River Tweed, UK. AB - This paper examines spatial and temporal variability in freshwater inputs of trace elements and major ions to the tidal reaches of the River Tweed. The relationships between concentrations of major ions and trace elements (in dissolved and acid-available particulate forms) and flow are examined, and a simple two-component endmember mixing analysis performed to link river water chemistry to catchment sources, and to identify whether within-river processes modify concentrations to produce non-conservative behaviour. The results indicate that most dissolved major ions and trace elements behave conservatively in the lower reaches of the River Tweed, the variability in concentrations being dominated by hydrology and the existence of different high-flow and low-flow endmember runoff chemistries. This suggests that the variability in concentration of most dissolved trace elements and major ions in the lower Tweed can be modelled using simple mixing relationships. However, the relationships between pH, Ca, Mg and Gran alkalinity show pronounced non-conservative behaviour, indicating the importance of understanding within-river process for modelling these determinants. The non-conservative behaviour identified is related to biological controls and the resultant diurnal variations in pH which promote daytime removal of Ca from solution by precipitation of calcium carbonate (and corresponding removal of Mg by co-precipitation) during extreme low-flow conditions and algal blooms during the summer. PMID- 10847154 TI - Epilithic and planktonic leucine aminopeptidase activity and leucine assimilation along the River Tweed, Scottish Borders. AB - The microbial organic-nitrogen transformation variables, extracellular leucine aminopeptidase activity, and leucine assimilation, were determined at stone surfaces and in river water at six sites along 138 km of the River Tweed and also in a major tributary, the River Teviot. Sampling was on 3 days, representative of spring, summer and autumn. The variables, except for epilithic leucine assimilation, all showed significant downstream increase on at least 2 of the sampling days. Their values tended to be lowest in spring (early April). Aminopeptidase activity, leucine assimilation, microbial abundance, and pattern of downstream increase in the Tweed were shown to be broadly similar to in the River Swale, northern England. Microbial organic-nitrogen transformations and abundance in the Teviot tended to equal or exceed those in the lower Tweed, probably because of enrichment of the tributary by sewage-works discharges. PMID- 10847155 TI - Suspended sediment in the Rivers Tweed and Teviot AB - Suspended solids concentrations were monitored over a 3-year period from April 1994 to March 1997 at two sites on the River Tweed and one on the Teviot, a tributary of the Tweed. Values of the median suspended solids concentration ranged between 6.6 and 8.0 mg l(-1) for the three sites. The relationship between suspended solids concentration and flow varied both between years and by month, showing the influence of sediment supply and diatom growth. Estimates of the flux of suspended solids from the Tweed basin at the tidal limit ranged from 38,000 to 89,800 t year(-1), depending on the year. Converted into a yield, these values are at the lower end of the range of both published estimates and those for other LOIS rivers. PMID- 10847156 TI - Seasonal variability of salinity, temperature, turbidity and suspended chlorophyll in the Tweed Estuary. AB - Results are presented from a campaign of measurements that were undertaken to examine seasonal variability in physical and chemical fluxes and processes within the Tweed Estuary during the period September 1996-August 1997. The study utilised monthly surveys, each of approximately 1 week duration. This article interprets a subset of the salinity, temperature, turbidity [suspended particulate matter (SPM) levels] and chlorophyll a data. Measurements discussed here were obtained throughout the estuary during high-speed transects that covered the region between the tidal river and the coastal zone. Longitudinal distributions of surface salinity depended strongly on freshwater runoff. During high runoff the surface salinity was low and the freshwater-saltwater interface (FSI) was located close to the mouth. The reverse was true at times of low runoff. Salinity stratification was generally strong. During the surveys, river runoff temperatures ranged from approximately 2 to 18 degrees C and coastal waters (approximately 33 salinity) from approximately 6 to 15 degrees C. Turbidity was low throughout the campaign (SPM < 30 mg l(-1)). Because of rapid flushing times (one or two tides), turbidity tended to mix conservatively between river and coastal waters. Higher coastal turbidity was associated with stronger wind events, and higher fluvial turbidity with spate events. Suspended chlorophyll a levels were usually low throughout the estuary (typically < 2 microg l(-1)) and showed large spatial variability. Because of the rapid flushing of the estuary, it is hypothesised that it was not possible for several algal cell divisions to occur before algae were flushed to the coastal zone. A 'bloom' occurred during the May 1997 survey, when chlorophyll a levels reached 14 microg l(-1). Higher chlorophyll a concentrations at that time occurred at very low salinities, indicating that these waters and algae were largely fluvially derived, and may have resulted from increasing springtime solar irradiation. PMID- 10847158 TI - Dredging the Tyne: an institutional perspective on process management AB - The increase in the size of steam vessels, and consequent need for the considerable enlargement of navigation channels, meant the years at the turn of the 19-20th centuries were of pivotal importance in harbour construction. This paper draws on the surviving archives for the Tyne estuary in north-east England in illustrating the inter-relatedness of both the physical and political processes, by which such adaptation and improvement were achieved. PMID- 10847157 TI - Distributions and seasonal variability of pH and alkalinity in the Tweed Estuary, UK. AB - Monthly axial profiles of the Tweed Estuary were carried out between July 1996 and August 1997 as part of the UK Natural Environment Research Council Land Ocean Interaction Study. During these surveys, master variables and a range of chemical constituents, including pH and total alkalinity, were measured between the mouth of the estuary, at Berwick-upon-Tweed, and the freshwater tidal reach, at Union Bridge, approximately 10 km up-estuary. Alkalinity and pH showed clear seasonal variations in values within the lower salinity region of the estuary, and these could be related largely to changes in river flows, and hence to variations in the ratios of the source waters. River waters at high flows were dominated by poorly buffered surficial runoff, and pH and alkalinity values were low; conversely at low flows, groundwaters rich in weathered bedrock ions dominated the river water, and pH and alkalinity values were significantly higher. Biological production and respiration also had an effect on this seasonal variability, although this was secondary. The behaviour of pH and alkalinity within the estuary appeared generally conservative, although apparent non conservative distributions were observed in the freshwater tidal reach. In a number of cases this behaviour could be attributed to biological activity. Estimated values of the excess partial pressures of CO2 within the fresh-water tidal reach were low (< 4), reflecting the relatively pristine character of the Tweed. During the spring and summer, CO2 was significantly undersaturated with respect to the atmosphere; extensive production by bed-anchored macroalgae was probably the reason for this, although the effects of water column algae cannot be discounted. In winter, production was minimal and respiration more important. As a result, partial pressures within the water column increased to a maximum of approximately four times atmospheric. Only infrequently was the CO2 in equilibrium between the river and atmosphere, suggesting that the tidal reach of the Tweed is a dynamic environment with respect to carbon cycling processes. PMID- 10847159 TI - The water quality of the River Wear, north-east England AB - A 1-year detailed study of water quality in the River Wear in combination with longer (decadal) term Environment Agency data (Harmonised Monitoring Scheme) show the influences of historic lead-zinc and coal mining and sewage inputs. The water quality for many determinands, such as sodium, chloride, boron, nitrate, and soluble reactive phosphorus, varies seasonally due to changing flow conditions. For most dissolved determinands, concentrations decrease with increasing flow in response to dilution of point and diffuse sources by rainfall. However, concentrations increase with increasing flow for dissolved organic carbon, aluminium, lead, iron, yttrium, and the lanthanides and actinides. This increase probably reflects two processes. Firstly, trace element-enriched runoff occurs from the acidic moorland areas of the catchment when it wets up. Secondly, at high flows, increased production of transition metals bearing microparticulate material occurs; these pass through conventional filters used to separate dissolved from particulate materials. For the particulate components, iron, manganese, aluminium, and some trace transition metals are present above the analytical detection limits and concentrations increase with increasing flow in response to increasing suspended sediment levels. However, particulate metal concentrations are relatively low for the Wear compared to the other eastern UK rivers, even though historic lead-zinc mining activity in the upper portions of the Wear catchment has led to extensive spoil contamination of the land. This feature probably reflects the lack of a significant flood plain area, where trace contaminants can accumulate within the sediments. The importance of inputs of pumped water from former coalmines is highlighted; increased manganese and dissolved carbon dioxide concentrations and reduced pH result. Examination of the Harmonised Monitoring Scheme data indicate no clear long-term changes in water quality for all but one determinand, although analysis is hampered by changes in sampling frequency for all determinands and changes in detection limits for certain trace metals. The only clear long-term change is a reduction in the micro organic insecticide, aldrin. Nonetheless, the long-term data, when plotted as a function of month, follow the same pattern as the LOIS data. For aldrin, the higher values during the early period of record occurred in the latter half of each year, presumably as a consequence of seasonal application and enhanced leaching during the autumn. PMID- 10847160 TI - Identifying sources of dissolved organic carbon on the River Swale, Yorkshire AB - A dissolved organic carbon (DOC) sampling study was carried out on the River Swale in Yorkshire as part of the NERC LOIS programme. Loads of DOC were calculated from the river and its tributaries. For the River Swale calculated loads varied between 1900 and 9000 kg/day and for the tributaries between 1 and 8500 kg/day. No relationship was found between the amount of organic carbon in the soil of the catchments and the DOC loads, perhaps because of the preceding dry summer. DOC loads were also estimated for sewage point sources and ranged between 7 and 287 kg/day. Some of the catchments draining to the River Swale respond with high DOC loads during high flow conditions, others do not. Some catchments are dominated by sewage point sources during low flows and diffuse sources during high flows. A mass balance for DOC on the River Swale was calculated, however, loads from inputs did not balance with outputs because of a lack of data and errors inherent in some of the approaches used. Despite this, results indicate in-stream losses of approximately 20%. A highly detailed measurement study of a stretch of river is recommended to identify all the sources and sinks of DOC, the relative size of their contribution and how they respond in differing conditions. PMID- 10847161 TI - Downstream change in leucine aminopeptidase activity and leucine assimilation by epilithic microbiota along the River Swale, northern England. AB - Parallel determinations of epilithic extracellular leucine aminopeptidase activity and leucine assimilation were made at five sites along 112 km of the River Swale and also in two tributaries, the River Wiske and Cod Beck. Epilithic leucine aminopeptidase activity along the Swale increased with distance downstream; this increase was gradual, rather than stepwise in response to specific sewage-works outfalls. Epilithic leucine assimilation, in contrast, did not consistently increase along the river. Epilithic leucine aminopeptidase activity and leucine assimilation were both potentially controlled by epilithic microbial variables (bacterial abundance and chlorophyll a) while leucine aminopeptidase activity was also strongly related to water-quality variables, especially temperature, pH and conductivity. Epilithic leucine aminopeptidase activity and leucine assimilation were coupled, but the magnitude of aminopeptidase activity was always substantially greater than that of leucine assimilation. Arguments are presented, however, which suggest that this did not necessarily indicate the constant availability of excess leucine, and by inference amino-acid nitrogen, to epilithic bacteria. Values of epilithic leucine aminopeptidase activity and leucine assimilation, expressed relative to rates in overlying water, suggested that most activity and assimilation was epilithic rather than planktonic, although the planktonic contribution was proportionately greater at the deeper, more downstream, sites. In the tributaries, River Wiske and Cod Beck, values of epilithic leucine aminopeptidase activity and epilithic microbial abundance, as well as those of many water-quality variables, resembled values in the middle and lower Swale. Thus, these tributaries were essentially lowland, enriched watercourses being very different from the headstreams of the main river. PMID- 10847162 TI - The particle size characteristics of fluvial suspended sediment in the Humber and Tweed catchments, UK AB - This paper presents information on the absolute (chemically-dispersed) particle size characteristics of the suspended sediment transported by rivers in the Humber and Tweed basins during the period 1994-1998. For most of the rivers, > 95% of the suspended sediment load at the time of sampling was < 63 microm (i.e. silt- and clay-sized material) and < 5% was > 63 microm (i.e. sand-sized material). The < 2 microm fraction (i.e. clay-sized material) typically ranged between 15 and 25%. The average median (d50) particle size for the individual rivers ranged between 4.1 and 13.5 microm. Generally, the particle size characteristics of suspended sediment transported in the two basins were similar. There were, however, noticeable spatial variations in the particle size composition of suspended sediment within the study basins, which reflected the particle size of the sediment sources and their spatial variation, and the selectivity of the sediment mobilization and delivery processes. When particle size parameters were plotted against discharge, there were no significant relationships, although there was some evidence of trends varying between sites. The lack of significant relationships with discharge reflects the fact that sediment particle size is largely supply-controlled, rather than a function of flow and hydraulics. When particle size variations were examined during individual storm events, there was evidence of a pulse of coarse sediment on the rising limb of the hydrograph. This may reflect the remobilization of coarse channel bed sediment as flow velocity and shear stress increase. Finer sediment was transported subsequently during the hydrograph peak and on the falling limb. The findings reported have important implications for understanding and modelling suspended sediment, and associated contaminant, dynamics in river basins. PMID- 10847163 TI - The influence of environmental factors on movements of lowland-river fish in the Yorkshire Ouse system. AB - The influence of several environmental parameters on movements of fish within the Yorkshire Ouse system, north-east England, is considered. Automated monitoring of fish with passive integrated transponding (PIT) tags, at the entrance to a fish pass on the lower Derwent, was used to examine factors influencing upstream migration of fish between May and August 1998. Overall, 85% of records were from the cyprinids, chub Leuciscus cephalus, dace Leuciscus leuciscus and roach Rutilus rutilus, which are the dominant species in the lower Derwent. Daily numbers of PIT-tagged fish entering the pass in summer were significantly and positively correlated with daylength, but not significantly correlated with mean daily temperature or mean daily flow. There were significant variations in the diel pattern of PIT records between species. Adult chub mostly approached the fish pass at night, while other species entered over a wider range of times, and stocked juvenile cyprinids entered mostly during daytime. Analyses of movements of radio-tracked barbel from the lower Nidd showed that downstream displacements associated with high flow in summer were often followed by homing to the original residence area, but that in autumn displacements were significantly more frequent and homing was significantly less frequent, resulting in a tendency for barbel to move downstream. It is concluded that an appreciation of the effects of environmental parameters on movements by different fish species is important in understanding the causation of spatial variations in distribution of fish in lowland rivers. PMID- 10847164 TI - Concentrations of suspended particulate organic carbon in the tidal Yorkshire Ouse River and Humber Estuary. AB - Data are presented for particulate organic carbon (POC) and particulate nitrogen (PN) concentrations in the Humber Estuary and tidal River Ouse Estuary. The POC data were derived from approximately monthly surveys and are consistent with data reported for suspended particulate matter (SPM) in the non-tidal River Ouse (the freshwater river) and with SPM, or bed sediments, in estuarine ecosystems such as the Mississippi, Delaware, San Francisco Bay, Tolo Harbour, the Vellar Estuary and Cochin Backwater, as well as the Loire, Gironde, Ems and Tamar Estuaries. Relative to the dry weight of SPM, the Humber-averaged organic carbon and nitrogen percentages during the year February 1995-March 1996 were 2.6 +/- 0.6% (mean and S.D.) and 0.21 +/- 0.04%, respectively. The ratio of Humber-averaged POC to Humber-averaged PN was 13 +/- 3. Higher POC levels were observed near the Humber's mouth and in the adjacent coastal zone during 'bloom' conditions, and in the upper estuarine reaches during large, winter and springtime freshwater inflows. At these times of high runoff, the POC content of SPM increased progressively up-estuary from the coastal zone to the tidal River Ouse. When inflows became very low, during late spring to early autumn of 1995, both the freshwater-saltwater interface (FSI) and the strengthening turbidity maximum (TM) moved further up-estuary and the POC content of SPM in the upper reaches of the Ouse became lower compared with that immediately down-estuary. This led to a poorly defined POC maximum near the confluence of the Humber, Ouse and Trent, before POC eventually decreased again towards the coastal zone. The lower POC contents in the upper estuarine reaches of the tidal Ouse may have been partly due to POC respiration by heterotrophic bacteria attached to SPM within the TM, consistent with the severe oxygen depletion observed there during high turbidity, summertime spring tides. PMID- 10847165 TI - Chlorobenzenes in rivers draining industrial catchments AB - Eleven chlorobenzenes (out of a total of 12 in the congener series) were monitored weekly on four industrialized rivers (Aire, Calder, Don and Trent) of the Southern Humber Catchment in whole water samples. 1,2- and 1,4 dichlorobenzene were present at relatively high levels on both the Aire and Calder, having mean concentrations of approximately 30 ng/l. They were both at lower concentrations on the Don and Trent, although the 1,4-isomer dominated. All other chlorobenzenes monitored were routinely found on all the rivers, with the exception of hexachlorobenzene, which was only regularly detected on the Trent. Again, the rivers fell into two classes with respect to their total chlorobenzene concentrations, with the Aire and Calder being more polluted. The higher levels of chlorobenzenes (excluding hexachlorobenzene which was used widely as a agricultural pesticide) on the Aire and Calder, and the dominance of the 1,4 dichlorobenzene congener (accounting for 60-70% of sigma chlorobenzenes) on the Don and Trent, indicated that the Aire and Calder were predominately contaminated with chlorobenzenes through industrial sources, while the Don and Trent were mainly contaminated through domestic sources (1,4-dichlorobenzene is widely used as a toilet deodorant). 1,4-Dichlorobenzene dominated flux, with the Aire, Don and Trent exporting 52.5 kg/year into the Humber estuary, followed by the 1,2 dichlorobenzene at 38.8 kg/year. Sigma chlorobenzenes exported to the Humber was 133 kg/year. This is the first study to calculate chlorobenzene fluxes to the North Sea from a UK catchment. PMID- 10847166 TI - Spatial and temporal regulation of the pesticide dieldrin within industrial catchments AB - The river catchments of south Yorkshire support a very high density of wool processing industries. Dieldrin was once used as a moth proofing agent, as a sheep dip, and as a pesticide to protect wool fleeces during storage and transport, all of which caused pollution of these catchments due to textile processing. Weekly sampling of four of these rivers revealed two classes of dieldrin contamination: the Aire and Calder (the rivers which support very high concentrations of wool processing industries) had higher concentrations (averaging approximately 3 ng/l) than the Don and Trent (approximately 1 ng/l). The average flux of dieldrin from these rivers into the Humber estuary was 9.8 g/day, with the Aire (of which the Calder is a tributary) and the Trent contributing almost equally, with a smaller contribution from the Don. The Trent has the highest average flow, explaining its large contribution to dieldrin flux. Less detailed sampling of rivers from the north Humber catchment which drain predominantly rural areas had dieldrin concentrations similar to the heavily industrialized southern catchment rivers. This suggests that dieldrin from agronomic and domestic usage may be more persistent than the pollution caused by textile processing industries. Evidence is presented to suggest that the principle dieldrin sources to the Humber catchments are sewage treatment plants, and that the dieldrin sources are in rapid equilibrium with the water column. PMID- 10847167 TI - Removal of linear alkylbenzene sulfonate from a small Yorkshire stream: contribution to GREAT-ER project #7. AB - An in-stream removal experiment has been carried out in Red Beck, a small stream which receives effluent from Shibden Head Sewage Treatment Works. This trickling filter works serves a purely domestic population of 9408 but is scheduled to be closed, and the flows diverted to another works, as part of Yorkshire Water's continuing capital investment programme. An anionic detergent, linear alkylbenzene sulfonate (LAS), boron, and standard water quality parameters have been measured at seven sites downstream of the effluent discharge point. Time of travel has been measured by detection of a fluorescent dye added to the effluent sampling chamber, and the increase in flow as the river proceeds through the catchment has been determined from current flow measurements, and from boron dilution data. Assuming a first order removal mechanism, the overall half-life for LAS removal is just over 2 h (2 h 14 min). Faster removal takes place in the upper portion of the stream, and removal over the last five sampling points is somewhat slower, with a half-life of 2 h 40 min. This removal may comprise both primary biodegradation and the deposition of suspended matter to which the surfactant has been adsorbed. There was no significant difference in the removal half-lives of the individual alkyl chain length homologues. PMID- 10847168 TI - Novel techniques for characterizing complex water use patterns within a network based statistical hydrological model AB - Information on the magnitude and variability of flow regimes at the river reach scale is a central component of most aspects of water resource and water quality management. Within the UK, river stretches with permanent gauging stations represent less than one percent of the total number of river stretches mapped at a scale of 1:50,000 and fewer than 20% of gauged catchments can be regarded as having natural flow regimes. This paper is the second of two papers that describe the development and application of hydrological models for estimating the variability and magnitude of natural and artificially influenced flow regimes at ungauged sites. The development of the models for estimating statistical descriptions of both the natural and artificially influenced flow regimes at ungauged river reaches is described by Young et al. (Young AR, Gustard A, Bullock A, Sekulin AE, Croker KM. Sci Total Environ 2000: this issue). This paper describes a pragmatic approach for applying the model to a complex river basin and characterizing the impacts of water use in the basin through example. The basin selected for the application contains the Aire and the Calder catchments within south Yorkshire. PMID- 10847169 TI - A river network based hydrological model for predicting natural and influenced flow statistics at ungauged sites: Micro LOW FLOWS AB - Information on the magnitude and variability of flow regimes at the river reach scale is a central component of most aspects of water resource and water quality management. Within the UK the number of river reaches that are gauged represent less than 1% of the total number of river stretches mapped at a scale of 1:50,000. The paper is the first of two papers that describe the development and application of procedures within Micro LOW FLOWS for estimating the variability and magnitude of flow regimes at ungauged sites as represented by flow duration and mean flow statistics. This paper describes the development of the models whilst Young et al. (this volume) describes a pragmatic approach for applying the model to a river basin with complex water use patterns. PMID- 10847170 TI - The use of measured boron concentration data from the GREAT-ER UK validation study (1996-1998) to generate predicted regional boron concentrations AB - The concentrations of boron in the Aire and Calder which have been measured in order to validate the GREAT-ER model have been used to derive a predicted environmental concentration (PEC) for boron in the Aire catchment in Yorkshire. The data have been obtained from 33 sites, with monthly sampling over a 2-year period. The 90th percentile of the means of measured concentrations (242 microg l(-1)) is higher than the 90th percentile predicted by the GREAT-ER model (186 microg l(-1)), if river stretches are normalised by their flow volumes, and unpolluted headwaters are included in the catchment averaging procedure. However, if the river stretches are normalised by the length of the respective stretch, and unpolluted headwaters are included, a lower regional PEC of 28 microg l(-1) is obtained. If normalisation by stretch length is appropriate for regional risk assessment, then the 90th percentile of the means of the measured data will overpredict the regional boron concentration in a catchment such as the Aire, with a low natural level of boron in the headwaters. Monitoring data for boron in the effluents from sewage treatment works in the Aire catchment, and for boron in the Went and Rother catchments, with a higher background boron level, are also given. PMID- 10847171 TI - Patterns in trace element chemistry in the freshwater tidal reaches of the River Trent AB - The major, minor and trace element chemistries at two freshwater tidal sites on the River Trent are described and compared with the non-tidal river, to demonstrate the variability in chemical processes in a major chemically and hydrologically active part of the river system, which is often overlooked in freshwater hydrochemical studies. The study shows a chemical gradient along which concentrations increase downstream for suspended sediments and acid-available particulate (AAP; > 0.45 microm) trace element fractions and also the 'dissolved' (< 0.45 microm) fractions of those trace elements with low-solubility phases. The study highlights the importance of tidally induced suspended-sediment dynamics and the generation of microparticulates for trace element transport within the tidal reaches. The microparticulate and particulate fractions are associated with both anthropogenic and lithogenous sources, and the concentrations of these two fractions co-vary according to a two-component mixing model involving riverine and tidal endmembers, with the tidal endmember exhibiting a strong lithogenous component. In many cases the AAP trace element concentrations are highly linearly correlated. At the downstream tidal site, suspended sediment concentrations were particularly high and ranged from 44 to over 24,000 mg/l as a result of tidally induced sediment mobilisation. As a consequence of this, particulate metal concentrations are especially high, and here, for the first time within the east coast studies of UK rivers, particulate fractions were measurable for trace elements such as boron and molybdenum, previously reported above detection limits only in dissolved form. It is demonstrated that the microparticulate components are not simply related to suspended sediment concentrations or to the bulk composition of the AAP fraction. Rather, microparticulate generation is related to a more complex pattern, probably linked to hydrodynamic factors involving sediment resuspension and microparticulate transport mechanisms. PMID- 10847172 TI - The water quality of the River Trent: from the lower non-tidal reaches to the freshwater tidal zone AB - The water quality of a major river system, the River Trent, which flows into the Humber estuary and drains large centres of population and industry in central England, is examined in terms of trace element, nutrient, major-ion, suspended sediment and carbon concentrations and set within the context of the transition from non-tidal to the tidal freshwater reaches. Detailed investigation of the water quality variability in the non-tidal river Trent has revealed mainly simple patterns in dissolved chemical concentrations, controlled largely by hydrology and the mixing of baseflow and stormflow endmember chemistries. However, silicon, carbon and nitrate show much more complex behaviour, as concentrations of these chemical determinands are regulated by in-stream biological processes. Major increases in concentrations of suspended solids and acid-available particulate (AAP) trace elements are shown to be linked to tidal mobilization of sediment and the location of the 'turbidity maximum'. Reductions in the concentrations of AAP trace elements per gram of suspended sediment suggest the mobilization of a different sediment type with a lower trace element content, which may result from: (i) a secondary sediment source; or (ii) mobilization of coarser-grained sediment under the stronger tidal hydrodynamic forces. Increased correlations for AAP-trace element interrelationships downstream suggest increasing simplicity of mixing relationships. PMID- 10847173 TI - A spatial study of denitrification potential of sediments in Belfast and Strangford Loughs and its significance. AB - The C2H2 inhibition technique was employed to study seasonal denitrification potential rates in sediment slurries from tidal and subtidal sites in Belfast and Strangford Loughs, Northern Ireland. A comparison of denitrification rates obtained from this method with those obtained from the 15N-gas flux method generally showed good agreement. Depth profiles measured up to 1 m showed that denitrification decreased with depth, with highest values in the 0-5-cm fraction. For the Belfast Lough tidal system a multiple regression model was developed which explained 83% of the variation in denitrification potential. The independent variables were water content, sediment temperature, total oxidizable N in porewater and total organic N. The highest rate of denitrification potential, 2100 micromol N m(-2) h(-1), was found in areas where there was a high anthropogenic input of nutrients. Denitrification in sediments in both loughs can play a potentially significant role in removal of NO3- from the overlying water. In Belfast Lough the overall denitrification potential rate matched the external NO3-N inputs, whilst in Strangford Lough it exceeded it by sixfold, which suggests a potential to remove future additional anthropogenic inputs to the Lough. PMID- 10847174 TI - Solid-solution metal partitioning in the Humber rivers: application of WHAM and SCAMP AB - The solid-solution partitioning of five trace metals (Co, Ni, Cu, Zn and Pb) in four LOIS rivers has been modelled using a predictive chemical speciation code (WHAM-SCAMP). Observed (log K(D,obs)) and predicted (log K(D,pred)) log K(D) values were similar for Zn. For Co and Ni, the log K(D,pred) values were typically greater than the log K(D,obs) values, while for Cu and Pb the reverse was seen. Removing modelled competition by Ca and Mg for binding sites on particulate organic matter increased the log K(D,pred) values for all the metals except Pb, and gave better agreement between the observations and the predictions for Co, Ni and Zn. Modelling solution iron as iron oxide particles decreased the log K(D,pred) values for Pb. The Cu predictions were very sensitive to the metal binding strength of dissolved and particulate organic matter. Overall, the model shows promise for the prediction of solid-solution metal partitioning in aquatic systems. The modelling exercise has identified the following uncertainties: the extent of Ca and Mg competition for binding sites, the chemical nature of the measured particulate metal, the efficacy of the solid-solution separation method and the strength of copper binding to organic matter. PMID- 10847175 TI - Suspended chlorophyll in the River Nene, a small nutrient-rich river in eastern England: long-term and spatial trends. AB - The distribution of suspended algae was investigated in a 69-km length of a small lowland river in the UK, the Nene, using chlorophyll as a measure of biomass. Variations in chlorophyll data collected between 1975 and 1996 by water management organisations at a downstream site (km 91.7) were evaluated against a range of physical and chemical variables. Interpretation was aided by additional sampling at eight sites on the main river (including km 91.7) and three tributaries between 1994 and 1996. Significant inter-year variation was evident in all data and was most pronounced in discharge. The latter half of the 22-year period had significantly higher temperature and light and significantly lower ammonia concentrations. Discharge, temperature and light were significant predictors of chlorophyll concentration, particularly between January and June, and spring chlorophyll maxima ranged from 106 to 276 microg l(-1). Summer chlorophyll concentrations were generally low relative to spring peaks. Some summers, however, had very low chlorophyll concentrations (average < 10 microg l( 1)) which were independent of the controlling factors at other times and inter year variation in submerged macrophyte abundance is proposed as a causal factor for their occurrence. Spring chlorophyll peaks occurred earlier and had smaller amplitude at downstream sites compared to those further upstream. Average spring chlorophyll concentration (April-June) increased significantly between km 22.4 and 43.9, thereafter remaining high to km 91.7. Spatial trends were attributed to changes in channel morphology, retention time and longitudinal variations in velocity and temperature. PMID- 10847176 TI - The water quality of the Great Ouse AB - The results of a 1-year detailed water quality study of the Great Ouse are presented. Being recharged to an important degree from groundwater sources within the carbonate bearing bedrock, the waters are enriched in base cations and they are of relatively high alkalinity (approx. 4000 microEq./l) and pH (approximately 8). Many chemical constituents such as sodium, chloride, boron, nitrate and soluble reactive phosphorus vary over time in response to changing flow conditions. For most determinands, concentrations decrease with increasing flow in response to dilution of point and groundwater sources by rainfall. However, for barium and nitrate, concentrations increase with flow and this is indicative of increased runoff from the agriculturally impacted soils. While this feature is certainly expected for nitrate due to fertiliser application, the pattern for barium is at face value unexpected as it would normally be expected to behave like other divalent base cations such as calcium, coming from aquifer leaching. However, it seems that agricultural disturbance of the land can also lead to enhanced runoff: a feature now becoming apparent in several UK studies. The concentrations of micro-organic herbicides vary over time in response to the different times of application and only isoproturon shows a clear response to varying flow conditions. With regards to biological processes, this is manifest by changes in pH, dissolved carbon dioxide and silica. At most times of the year the waters are oversaturated with respect to atmospheric CO2 (EpCO2) by a factor of approximately 6 and at these times pH is approximately 7.7 and silica concentrations are approximately 4 mg-Si/l. However, in the early spring period pH increases to values over 8, EpCO2 declines to about the atmospheric levels and silica declines to approximately 1 mg-Si/l. This change probably reflects the dynamic diatom blooms and decays common in this river. PMID- 10847177 TI - The water quality of the River Thames at a rural site downstream of Oxford AB - Water quality information is presented for the River Thames 34 km downstream of the market town of Oxford in Oxfordshire to provide an overview of the hydrochemical functioning of a major agriculturally impacted river entering the North Sea. The data, which cover the period from the spring of 1997 to the spring of 1999, relate to three types of data. These types are: (1) weekly spot sampling for determination of major, minor and trace elements, pH, alkalinity and herbicides; (2) tri-weekly spot sampling for pH, alkalinity and dissolved silicon; and (3) continuous measurements of pH and dissolved oxygen. Calcium and bicarbonate provide, respectively, the dominant cation and anion in solution and their compositions remain relatively constant through time, irrespective of flow levels. In contrast, many determinands show seasonally related fluctuations. Concentrations for most of the major anions, sodium, potassium as well as soluble reactive phosphorus and several soluble trace elements such as boron, antimony, arsenic and molybdenum decrease as flow increases. A reverse pattern is observed for nitrate, some herbicides and trace elements associated with particulate phases: concentrations increase with increasing flow. These patterns reflect the influence of: (1) a calcium carbonate rich groundwater system which provides the main stream flow component; (2) dilution of point source pollutant inputs associated with sewage and possibly light industry at high flows for several major, nutrient and trace elements; (3) enhanced nitrate and herbicide runoff from agricultural land at high flows; (4) enhanced microparticulate trace metal levels associated with increased suspended sediment loads at high flows; and (5) biological processes which affect pH, dissolved silicon, dissolved carbon dioxide and dissolved oxygen levels. An examination of soluble reactive phosphorus (SRP) and boron relationships reveals a reduction in concentrations for SRP associated possibly with phosphorus removal from a major sewage treatment works on an upstream tributary of the Thames, the Thame. PMID- 10847178 TI - The water quality of a tributary of the Thames, the Pang, southern England. AB - The water quality of a tributary of the Thames, the Pang, draining a rural part of the Thames basin is described and related to the influences of inputs from farming activity and groundwater from the underlying chalk aquifer as modified by biological within-stream processes. The groundwater inputs ensures that the waters are calcium and bicarbonate bearing and have relatively uniform concentrations. Agricultural inputs result in enhanced levels of nutrients, nitrate and soluble reactive phosphorus (SRP) in particular. For nitrate, the concentrations are higher during the winter months due to increased surface runoff. In contrast, SRP shows a more erratic behaviour with higher concentrations occurring during the early storm hydrograph rise following summer baseflow recession. Within the stream, biological activity results in: (a) marked diurnal fluctuations in the dissolved levels of carbon dioxide and oxygen; (b) a strong seasonal pattern in chlorophyll a levels; and (c) dissolved silica concentration reductions during the early spring period. Carbon dioxide levels are particularly high in the groundwater (typically approximately 60 times the atmospheric value) as observed at a spring discharge. However, within the stream, considerable degassing occurs although values remain an order of magnitude above atmospheric pressure. The findings are discussed in the context of the water quality functioning of agriculturally and sewage impacted southern eastern UK rivers. For example, the work shows that unlike for riverine systems with point sewage discharges or limited groundwater storage, there is a very poor link between chemical concentrations and flow even for components such as SRP and boron which are often connected to sewage discharges. PMID- 10847179 TI - The water quality of the River Kennet: initial observations on a lowland chalk stream impacted by sewage inputs and phosphorus remediation. AB - The water quality of seven sites on the upper reaches of the River Kennet round the market town of Marlborough is described and related to the introduction of phosphorus treatment of effluent from Marlborough sewage treatment works (STW). The River Kennet is mainly groundwater-fed from a Cretaceous chalk aquifer and hence the river water is calcium- and bicarbonate-bearing and has a relatively constant composition of many major water quality determinants. In-stream biological activity gives rise to marked diurnal fluctuations in pH (of approx. 0.8 units). Dissolved carbon dioxide and dissolved oxygen also show marked diurnal fluctuations. Dissolved carbon dioxide varies from approximately 10 to 70 times atmospheric pressure, indicating net release of carbon dioxide and the dominance of heterotrophic (respiratory) processes over autotrophic processes (photosynthesis). Much of the excess carbon dioxide is probably associated with carbon dioxide laden groundwater inputs and the relatively short within-stream residence times ensures only limited degassing to the atmosphere. Diurnal fluctuations in dissolved oxygen vary from approximately 20% to 200% saturation. For both dissolved carbon dioxide and dissolved oxygen, the amplitude of fluctuations is much lower during the winter period, when biological activity is at its lowest. The concentrations of soluble reactive phosphorus (SRP), total phosphorus (TP) and boron increase markedly just downstream of the sewage works as a result of this point source input. These concentrations slowly decline further downstream as additional groundwater inputs dilute the effluent further. The introduction of chemical treatment of sewage effluent for phosphorus reduction at Marlborough STW resulted in a marked decrease in within-river SRP and TP concentrations to levels approximately the same as those upstream of the STW. A comparison of SRP and boron concentrations reveals a reduction in in stream SRP concentrations by approximately 75% following effluent treatment. In terms of within-river processes controlling in-stream phosphorus concentrations, previous studies have indicated that one potentially important mechanism within calcium bicarbonate bearing rivers may be related to co-precipitation of phosphorus with calcium carbonate (calcite). The present study shows that the waters are oversaturated with respect to calcium carbonate, that no equilibrium conditions exist and that phosphorus removal has led to undetectable changes in calcium carbonate oversaturation. Hence, it is concluded that the primary changes in phosphorus levels within the river is directly associated with changing point source contributions from the STW and physical dilution within the river. However (1) the results relate to only the first year of study and subsequent differences may become apparent and (2) reactions between the water column and plant and bottom sediment interfaces may be important in regulating phosphorus fluxes within the river. The results presented in this paper mark a pilot phase of a longer-term initiative and this paper provides a background setting. The paper discusses the longer-term objectives and important gaps in knowledge of the system that requires further address. PMID- 10847180 TI - Temporal and small-scale spatial variations of dissolved oxygen in the Rivers Thames, Pang and Kennet, UK. AB - Small-scale spatial dissolved oxygen concentrations were measured on three rivers by taking a number of transects along a river reach. Temporal (diurnal) variations were measured by use of continuous (15-min) monitors located at the mid-transect of the spatial observations. Spatial variation of dissolved oxygen concentrations were found to be relatively small (approx. +/- 10% dissolved oxygen) and biologically insignificant compared to diurnal changes under conditions of high productivity (approx. +/- 60% dissolved oxygen). The diurnal curves were analysed in order to estimate values of reaeration, photosynthesis and respiration on a daily basis for almost 3 months using process-based analytical techniques (the Delta and Hornberger and Kelly methods). The River Kennet was shown to be the most productive river followed by the River Pang and then the River Thames. The patterns of reaeration, photosynthesis and respiration and the behaviour of the three rivers are discussed. PMID- 10847181 TI - Phosphate measurement in natural waters: two examples of analytical problems associated with silica interference using phosphomolybdic acid methodologies AB - Assessment of phosphate species for bicarbonate-bearing lowland rivers of the Thames valley, England and acidic Welsh upland streams (the Plynlimon area, mid Wales) reveal problems of silica interference when employing standard automated 'phosphomolybdic acid' colorimetric methodologies. The interference occurs under a combination of two conditions: (1) when the reagents and samples are heated to speed up the rate of formation of phosphomolybdic acid; and (2) when the strength of a sulfuric acid reagent is too low. While this paper alerts researchers to potential analytical problems with silica interference within phosphomolybdic acid methodologies to determine soluble reactive phosphorus (SRP) concentrations, it cannot detail whether or not there is a general problem. This lack of specificity occurs because there are a variety of phosphomolybdic acid methodologies available which vary in their reagent recipes and it is not clear which methods will or will not suffer from silica interference. Changing the sulfuric acid reagent strength by a factor of two overcame the problem, in this case, with regards to the determination of soluble reactive phosphorus. However, even here, the method may not be applicable to very high alkalinity waters owing to their potential for reducing the acidity of the analyte. With regard to total dissolved phosphorus (TDP) and total phosphorus (TP) measurements, the importance of undertaking the analysis under ambient conditions is clearly shown. There are many small variants on standard phosphomolybdate methods for determining SRP, TDP and TP in use and it is not always clear which methods will or will not show silica interference for particular water types. It is therefore recommended that individual laboratories check their methodologies for silica interference using phosphate-free solutions with similar silica and alkalinity ranges for the waters being assayed. PMID- 10847182 TI - Characterising phosphorus loss in surface and subsurface hydrological pathways AB - The magnitude and composition of the phosphorus (P) load transported in surface and subsurface hydrological pathways from a grassland catchment depends on the discharge capacity of the flow route and the frequency with which the pathway operates. Surface runoff is an important pathway for P loss, but this pathway is spatially limited and temporarily confined to high magnitude, high intensity rainfall events. High P concentrations (mean: 1.1 mg TP l(-1)) were recorded, with most P transported in the dissolved fraction. Preferential flow pathways, particularly soil macropores and field drains, are important contributors to the overall P load; most P is transported in the particulate fraction and associated with organic or colloidal P forms. High P concentrations (mean: 1.2 mg TP l(-1)) were recorded in macropore flow in the upper 0-15 cm of a grassland soil, and generally declined with increasing soil depth. On average, P concentrations in drainflow were over six times greater in stormflow compared to baseflow. Stormflow P losses in drainflow were predominantly in the particulate fraction; significant correlation (P < 0.01) was recorded with suspended sediment concentrations in drainflow. Phosphorus concentrations in groundwater were low (< 0.2 mg TP l(-1) at 150 cm), although this pathway may contribute to stream flow for the majority of the year. PMID- 10847183 TI - Assessing the potential impacts of various climate change scenarios on the hydrological regime of the River Kennet at Theale, Berkshire, south-central England, UK: an application and evaluation of the new semi-distributed model, INCA AB - A new semi-distributed integrated nitrogen in catchments (INCA) model was used to attempt to assess the potential impacts of several recent Hadley Centre climate change scenarios on the hydrological flow regime of the entire River Kennet catchment to Theale, south-central England, UK. The climatically and hydrologically anomalous period 1985-1995 was used for baseline data in an attempt to: (1) represent any possible future climatic or hydrological variability not available from scenario use alone; and (2) attain maximum possible model calibration validity under future climates by simulating extremes of within-year hydrological variability. Substantial reductions in total annual runoff occurred, with an average reduction of 18.97%. Summer and late autumn soil moisture deficits (SMDs) increased in intensity, and were also found to persist for longer periods into autumn and (occasionally) winter. A generally enhanced hydrological regime of the River Kennet was simulated, with increased seasonality overall. A greater percentage of flow was observed to occur in spring and (occasionally) winter. Month-to-month variability of flow was discovered to be greater than annual changes. An average reduction in minimum annual flows of 46.03% occurred. Implications for catchment ecology and water resource requirements are briefly discussed. An evaluation of the new INCA model's performance as a tool for climate change impacts assessment is made. PMID- 10847184 TI - Conclusions to the special issue of Science of the Total Environment concerning 'The water quality of UK rivers entering the North Sea'. AB - This paper presents an overview of the water quality functioning of eastern UK rivers draining into the North Sea. It summarises the results of three special issues of Science of the Total Environment and related work and complements a companion bibliography (Neal and Turner, 2000, this volume). The rivers are described in straightforward broad terms, and the reader is guided via the bibliography to the special volumes for the detailed findings. The water quality is shown to be highly variable over space and time. The changes over the past 300 years reflect first the transition from a rural society to the major impact of the industrial revolution and subsequently to one of the worlds' most successful environmental cleanups. The present water quality reflects the relative impacts of diffuse agricultural sources, urban and industrial point source discharges together with background inputs associated with geology: all these can be modified by within-river processes associated with physical mixing, biology and inorganic reactions. The work is set within the context of water quality, modelling and flux outputs to the North Sea and contemporary and future environmental research and management needs. A range of environmental impact studies is included. The detailed information is now available on a major and easily accessible database. PMID- 10847185 TI - A bibliography for the conclusions to the special issue of Science of the Total Environment concerning 'The water quality of UK rivers entering the North Sea'. Sci. Tot. Environ., this volume. AB - In this document, a bibliography for the conclusions to the special issue of Science of the Total Environment concerning 'The water quality of UK rivers entering the North Sea'. Sci. Tot. Environ., this volume, is presented. It comprises a list of the papers presented in five special issues in learned journals, three in Science of the Total Environment (this volume and volumes 194/195, 1997 and 210/211, 1998) as well as Hydrological Processes (1999, vol 13: Special Issue, River Basin Sediment Dynamics, eds. Anderson, M.G., Peters, N.E. and Walling, D.E.) and Marine Pollution Bulletin [1999, vol 37(3-7) Special Issue: Flux of Materials between Rivers and Coastal Waters, eds. Stebbing, A.R.D., Huntley, D. and Leeks, G.J.L.]. The bibliography comprises six main sections with subsections for specific water quality issues. These are as follows: Section 1 deals with the introduction to LOIS and contained in this section is background material over the nature of the programme, the study area, remit, water quality issues and international context. Section 2 provides the main meat to the bibliography and it splits into Section 2.1 historical and Section 2.2 current water quality issues. Within Section 2.2 there are subsections dealing with specific current aspects of water quality (Section 2.2.1, major ions; Section 2.2.2, pH, dissolved carbon dioxide and dissolved oxygen; Section 2.2.3 nutrients; Section 2.2.4, trace elements; Section 2.2.5 micro-organics; Section 2.2.6 sediments; Section 2.2.7 biology). Section 3 deals with hydrology and climate variability, Section 4, modelling, Section 5 load estimation and Section 6 conclusions. A full listing of the LOIS programme is available from the LOIS programme office, CCMS, Plymouth Laboratory, Prospect Place, West Hoe, Plymouth, PL1 3DH, UK (e-mail lois@ccms.ac.uk). PMID- 10847186 TI - A summary of river water quality data collected within the Land-Ocean Interaction Study: core data for eastern UK rivers draining to the North Sea AB - A numerical summary of the water quality of rivers draining into the North Sea from the eastern UK is presented using core information collected within the Land Ocean Interaction Study (LOIS) and a companion study by the Institute of Hydrology. The analysis is based on weekly monitoring for periods from 1993 to 1999 for major, minor, nutrient, trace and other water quality determinand chemistry. The data cover rivers ranging from the rural Tweed in southeastern Scotland, to the urban and industrially impacted Wear and Humber rivers in the north and central England and two agriculturally impacted rivers in the south of England (Great Ouse and Thames). Within the analysis, monthly averaged concentrations are plotted to show the seasonality. The summary provides specific information on the water quality of UK rivers which is of use in developing European and global initiatives for assessing pollutant inputs to estuarine, coastal and open-sea environments. PMID- 10847187 TI - Electrical impedance tomography (EIT) in applications related to lung and ventilation: a review of experimental and clinical activities. AB - This review article is a summary of the publications dealing with the pulmonary applications of electrical impedance tomography (EIT). Original papers on EIT lung imaging published over 15 years are analysed and several aspects of the performed EIT measurements summarized. Information on the type of the EIT device and electrodes used, the studied transverse thoracic planes, the data acquisition rate, the number of studied animals, normal subjects or patients, the kind of lung pathology, the performed ventilatory manoeuvres and other interventions, as well as the applied reference techniques, is given. The type of the generated pulmonary EIT images and the quantitative analysis of the EIT data are described. Finally, the major results achieved are presented, followed by an analysis of the perspectives of EIT in clinical applications. A comparative analysis of the EIT hardware and the quality of the evaluation tools was not performed. PMID- 10847188 TI - Non-invasive assessment of fluid volume status in the interstitium after haemodialysis. AB - During dialysis excess fluid is removed from uraemic patients. The excess fluid is mainly located in the skin and subcutaneous tissues. In this study we wished, with two noninvasive techniques, the IM (impression method) and BIA (bioimpedance analysis), to study what mechanical (IM) and electrical cellular membrane (BIA) effects the fluid withdrawal has on these tissues. The IM measures the resistive force of the tissues when mechanically compressed. From the force curve two parameters are calculated, the F(0), indicative of interstitial tissue pressure and the FT corresponding to the translocation of tissue fluid (interstitial movable water). The BIA phase angle shift (phi), i.e. geometrical angular transformation of the ratio between reactance and resistance, which has been associated with cellular membrane function, was used as a measurement of electrical cellular membrane effects. Twenty patients were studied before and after haemodialysis measuring the F(0), FT and phi. Theresults showed that the patients lost a median of 3.7 kg during the haemodialysis. F(0) increased until after dialysis, but did not reach significant values, whereas FT increased significantly after dialysis, p < 0.001, as compared with before. After a peak at one hour postdialysis the FT value returned to predialysis values at four hours after termination of dialysis. Also phi increased from before to after dialysis, p < 0.001, but already after one hour it returned to predialysis values. It is common knowledge that dialysis alters the dynamics of fluid in the interstitium of the skin and subcutis. We conclude that the impression method is sensitive enough to detect and chronicle these changes. Furthermore, with the BIA, (phase angle) signs of changes in the electrical properties of the tissues, possibly reflecting cellular membrane function, could be detected. PMID- 10847189 TI - Tracing air flow and diagnosing hypopnoeas in normal subjects. AB - Hypopnoea is a type of sleep-related breathing disorder (SRBD), and the apnoea plus hypopnoea index (AHI) is usually computed to diagnose this condition. We introduce a new method to diagnose flow with internal thermistors located on the same sensors as we use to diagnose obstructive segments in patients with sleep related breathing disorders. The aim of this study is to investigate whether internal thermistors are reliable for diagnosis of hypopnoeas. Fifteen volunteers participated in a prospective comparative study in healthy subjects without SRBD. We simultaneously measured minute ventilation by a pneumotachograph and indirectly air flow by internal thermistors in awake subjects was manually analysed from both the pneumotachograph and the internal thermistors. We found a close agreement between the different methods for percentage reduction in air flow both for the hypopnoea and the apnoea data. The mean difference between the percentage reduction in flow from normal breathing to hypopnoea measured by the pneumotachograph and the internal thermistors lying supine was 3.8% (SD 7.4). In the lateral position the corresponding figures were 1.0% and 4.6. This study in awake, normal subjects indicates that internal thermistors are as reliable as the pneumotachograph in diagnosing hypopnoeas and we believe that the reliability of this monitoring method is adequate for clinical use. PMID- 10847190 TI - Time-frequency analysis of heart rate variability using short-time fourier analysis. AB - This study was done to introduce new parameters derived by time frequency analysis of heart rate variability data. Four simulation experiments were carried out to compare the short-time Fourier transform (STFT) analysis method to the traditional overall spectral analysis method. Sinusoidal signals were generated with identical total power in the high-frequency band, but varying time-frequency and time-amplitude information. The STFT method was also applied to heart rate variability data from the stages of normal human sleep. Data analysis included computation of the power in the high-frequency band by overall spectral analysis. The instability coefficients (ICs) of the frequency and power in the high frequency band were derived by STFT analysis. The ICs derived by the STFT method were able to describe time-frequency and time-amplitude variations in sinusoidal signals which contained identical total power in a specified frequency range. The ICs of the frequency and power were able to differentiate variations in vagal activity between the stages of human sleep and waking. The ICs represent new parameters derived by the STFT method, and allow the detection and quantification of short-lasting time-frequency and time-amplitude variations that remain obscured by overall spectral analysis. PMID- 10847191 TI - Effect of increasing blood flow on distribution of pulmonary transit times in man. AB - The architecture of the vascular network is a major determinant of the distribution of transit times in the organ. Changes in pulmonary blood volumes, pulmonary transit times and the heterogeneity (coefficient of variation) of pulmonary transit times in man were measured using the first-pass radiotracer technique both at rest and during muscular exercise. Dichotomously branching and dispersive flow models of the lung vasculature were applied. The results showed that total pulmonary blood volume increased by 29%, total transit time decreased by 55% and the heterogeneity of pulmonary transit times decreased by 31% when blood flow increased threefold. All changes obeyed power law functions. The changes were greatest when blood flow increased from baseline to intermediate levels. Further increases in total blood flow were associated with decreases in transit times but not with more homogeneous distribution of transit times. The simulated results suggest that the transit time through the arterial vessels and the large veins have a negligible heterogeneity. Most of the variability of transit times through the lungs is due to the heterogeneity of the transit times through the capillaries and venules. PMID- 10847192 TI - Measurement of extracellular fluid volume in the neonate using multiple frequency bio-impedance analysis. AB - Bioelectrical impedance analysis may be useful for cot-side monitoring of fluid balance in the neonatal intensive care unit (NICU). However the presence of cardio-respiratory monitoring equipment, non-ideal electrode placement and inability to obtain accurate crown-heel measurements may interfere with the ability to obtain reliable impedance data in this setting. This study aimed to investigate the effects of these factors on impedance analysis and to develop a prediction equation for extracellular fluid volume in the neonate. The study found that cardio respiratory monitoring had no significant effect on multi frequency impedance measurements. Placement of current delivering electrodes on the ventral rather than dorsal surfaces improved separation of current and voltage electrodes but did not alter impedance results. Contralateral measurements were not significantly different to ipsilateral measurements. In 24 infants, extracellular fluid volume was measured using corrected bromide space (CBS) and simultaneous impedance analysis was performed. There was good correlation between CBS and the impedance quotient FF2/Ro where F is foot length and R0 is resistance at zero frequency. The study concludes that despite many potential difficulties associated with impedance analysis in the NICU, reliable measurements of impedance can be obtained and further work to validate prediction equations for ECF is warranted. PMID- 10847193 TI - Standardized limb abduction for bioimpedance measurements using position restraints. AB - Bioelectrical impedance analysis (BIA) measurements are widely used in the assessment of body hydration. However, there are many variables which can influence the impedance values obtained and their individual significance is not entirely clear. One such variable is limb position and it is possible to standardize this either by invoking subject cooperation or by the use of some form of mechanical constraint. This study evaluates these two approaches. BIA measurements were made on five healthy male volunteers both with the positioners and without. Additional variables including room temperature, prandial status, exercise, alcohol intake, bed surface for the test, and bladder status were controlled. The mean percentage difference between impedance values with and without positioners was (-0.37 +/- 0.69)%. The mean difference between occasions with and without mechanical fixing was (2.02 +/- 2.5)% and (2.58 +/- 3)% respectively. None of these differences is statistically significant. It was concluded that reproducibility of BIA measurements obtained with active subject cooperation in limb position is not significantly improved if mechanical positioners are used. This may not apply where subject cooperation is poor e.g. in very sick or very young people or in repeated measurements in different days. PMID- 10847194 TI - A direct comparison of wet, dry and insulating bioelectric recording electrodes. AB - Alternatives to conventional wet electrode types are keenly sought for biomedical use and physiological research, especially when prolonged recording of biosignals is demanded. This paper describes a quantitative comparison of three types of bioelectrode (wet, dry and insulating) based on tests involving electrode impedance, static interference and motion artefact induced by various means. Data were collected simultaneously, and in the same physical environment for all electrode types. Results indicate that in many situations the performance of dry and insulating electrodes compares favourably with wet electrodes. The influence of non-stationary electric fields on shielded dry and insulating electrode types was compared to wet types. It was observed that interference experienced by dry and insulating electrode types was 40 dB and 34 dB less than that experienced by wet electrode types. Similarly, the effect of motion artefact on dry and insulating electrodes was compared to wet types. Artefact levels for dry and insulating electrodes were significantly higher than those for wet types at the beginning of trials conducted. By the end of the trial periods artefact levels for dry and insulating types were lower than wet electrodes by an average of 8.2 dB and 6.8 dB respectively. The reservations expressed in other studies regarding the viability of dry and insulating electrodes for reliable sensing of biosignals are not supported by the work described here. PMID- 10847195 TI - Determination of stroke volume by means of electrical impedance tomography. AB - ECG-gated electrical impedance tomography (EIT) is a non-invasive imaging technique, developed to monitor blood volume changes. This study is the first in comparing this non-invasive technique in measuring stroke volume with established techniques. The objective of this study was to validate EIT variables derived from the EIT images with paired obtained stroke volume measurements by thermodilution and MRI. After right cardiac catheterization, EIT measurements were performed in 25 patients. Regression analysis was used to analyse the relation between the EIT results and stroke volume determined by thermodilution. From the regression line an equation was derived to estimate stroke volume (in ml) by EIT. A strong correlation was found between EIT and stroke volume measured by the thermodilution method (r = 0.86). In a group of 11 healthy subjects this equation was validated to MRI. The mean and standard deviation of the difference between EIT and MRI was 0.7 ml and 5.4 ml respectively. These data indicate that EIT is a valid and reproducible method for the assessment of stroke volume. PMID- 10847196 TI - Measurement of high resolution ECG QT interval during controlled euglycaemia and hypoglycaemia. AB - During hypoglycaemia, typically there is a change in the surface ECG characterized by a flattened and prolonged T wave, often accompanied by a fused U wave. The QT interval is a useful parameter for quantifying the ECG morphology. However, reliable measurement of QT is not straightforward, particularly for hypoglycaemic ECG morphology. The objective of this study was to compare the ability of two methods of QT measurement to distinguish between ECGs recorded during euglycaemia and hypoglycaemia. The first method involves manually setting the intersection of the isoelectric line and the T wave or, where this is not possible, the nadir between the T and U wave. The second method is semi-automatic and fits a tangent to the point of maximum gradient on the downward slope of the T wave. Two independent observers used both methods to measure the QT for high resolution ECG data recorded during a study of 17 non-diabetic subjects undergoing controlled euglycaemia and hypoglycaemia. Using the mean results of the two observers, the mean +/- SD increase in heart rate corrected QT, QTc, for ECGs recorded during euglycaemia and hypoglycaemia was 32 +/- 25 ms for the non tangent method and 60 +/- 24 ms for the tangent method. Therefore, the tangent method provides greater distinction between ECGs recorded during euglycaemia and hypoglycaemia than the non-tangent method. A potential clinical application could be the non-invasive detection of impending hypoglycaemia at night, which would be of significant benefit to adults and young children with diabetes. PMID- 10847197 TI - Heart rate and blood pressure variability in normal subjects compared with data from beat-to-beat models developed from de Boer's model of the cardiovascular system. AB - The objective of this study was to assess the ability of de Boer's model of the cardiovascular system to reproduce the heart rate and blood pressure variability observed in a range of normal subjects, and to make modifications to improve its performance. ECG, blood pressure and chest wall movement were recorded from 12 normal human subjects during controlled breathing. For each beat, systolic pressure, diastolic pressure, arterial time constant and RR interval were extracted. RR interval and systolic pressure spectral power in low and high frequency bands and the baroreflex sensitivity index, alpha, were then determined. For each subject, mean values were input to the model and the beat-to beat output compared with the actual data for that subject. Finally, the effects of reducing the influence of baroreflex on peripheral vascular resistance and of providing separate sympathetic and vagal baroreflex sensitivities were assessed. Simulations resulted in data which were qualitatively similar to those of each subject's recording. With the modifications, the log ratio of simulated to real data improved from 7.2 to 1.5 (p = 0.003) for low frequency RR, from 0.27 to 0.55 (p = 0.011) for high frequency RR and from 8.5 to 0.9 (p = 0.003) for low frequency systolic pressure. We conclude that de Boer's model reproduces many of the characteristics of heart rate and blood pressure variability, and our modifications to baroreflex sensitivities and the feedback effect on peripheral resistance resulted in significant improvements. PMID- 10847198 TI - Spectral analysis of left ventricular area variability as a tool to improve the understanding of cardiac autonomic control. AB - Spectral analysis of the fluctuations in heart rate (HR) or blood pressure (BP) has been extensively used as a tool for the noninvasive assessment of autonomic control of the heart. The recently developed echocardiographic acoustic quantification allows noninvasive continuous measurement of the left ventricular cross-sectional area (LVA) signal. In this study, we investigated whether the LVA signal, and more specifically its fluctuations, can be reliably subjected to spectral analysis, and whether the results of such analysis may improve the understanding of the cardiovascular control mechanisms. Our results show that the general pattern of power spectra of LVA fluctuations, as well as their reproducibility, is similar to the power spectra of HR and BP fluctuations. Analysis of LVA signals obtained in normal subjects at rest as well as under vagal blockade and under held respiration, and in patients with known autonomic dysfunction, showed significant differences between groups and states. The effects of age, related to the reduction in parasympathetic activity, were not evident in the spectral content of the LVA and BP signals. The high frequency LVA fluctuations are mainly of mechanical origin, since they were eliminated by breath-holding. We observed an increase in the high frequency LVA fluctuations under vagal blockade, indicating that under normal (control) conditions, these high frequency fluctuations are attenuated by parasympathetic activity. The enhancement in high frequency fluctuations in LVA observed in diabetic patients can thus be attributed to reduced parasympathetic activity. The analysis of LVA variability may be used as a tool for basic research and, possibly, as a quantitative clinical measure for specific disease states. PMID- 10847199 TI - Characteristics of laser Doppler perfusion imaging in vitro and in vivo. AB - Traditional laser Doppler flowmetry (LDF) employs continuous recording of perfusion at one point with time. In order to eliminate the large spatial and temporal fluctuations that occur in the microcirculation, laser Doppler perfusion imaging (LDI) integrates flow readings over a large area. This paper describes a number of experiments to identify some of the characteristics of the LDI, its relationship to flow and no-flow conditions and to compare it with LDF. We undertook experiments to establish the effect of scanner head height, avascular skin thickness and haematocrit on LDI output. We also investigated the contribution of the biological zero signal (the signal obtained from skin when flow is arrested) to the LDI output. LDI output increased with scanner height in vitro and in vivo. Increasing avascular skin thickness reduced the LDI output although linear output characteristics with flow were maintained over the flow range studied. Increasing the haematocrit resulted in a loss of linearity of output with flow at lower velocities. The biological zero signal contributes a similar proportion of the output signal in LDF and LDI. We have presented a series of experiments that will contribute to the understanding of the characteristics of laser Doppler perfusion imaging, its comparison to laser Doppler flowmetry and its relationship to flow and no flow situations. However, our experiments were restricted to one machine, and may not necessarily be applicable to other instruments. PMID- 10847200 TI - Cryptic genetic variation in parasitic protozoa. PMID- 10847201 TI - Application of PCR to the identification of dermatophyte fungi. AB - Infection of the keratinised tissues (skin, hair and nails) in man and animals by keratinophilic fungi (dermatophytes) results in dermatophytosis (also known as tinea or ringworm). As conventional laboratory procedures for the identification of dermatophytes are either slow or lack specificity, improved diagnostic methods are required. The application of nucleic acid amplification technology has made rapid and precise identification of dermatophytes possible. Recent studies have shown that when one of the four random primers (OPAA11, OPD18, OPAA17 and OPU15) was used in arbitrarily primed PCR (AP-PCR), up to 20 of the 25 dermatophyte species or subspecies under investigation could be distinguished on the basis of characteristic band patterns detected in agarose gel electrophoresis. A combination of two random primers (OPD18 and OPAA17) used in separate reaction tubes identified 23 of the 25 dermatophyte species or subspecies examined. AP-PCR provides a rapid and practical tool for identification of dermatophyte isolates that is independent of morphological and biochemical characteristics and thus enhances laboratory diagnosis of dermatophytosis. PMID- 10847202 TI - Relationship between asymptomatic carriage of Streptococcus pyogenes and the ability of the strains to adhere to and be internalised by cultured epithelial cells. AB - This study was undertaken to determine whether the ability of group A streptococci to persist in the throat following antibiotic therapy corresponded with their capacity to adhere to and be internalised by epithelial cells. The study employed a HEp-2 cell model to examine the adherence and internalisation capacities of 42 strains (13 from asymptomatic patients with bacteriological eradication failure and 29 from patients with bacterial eradication). The adherence and internalisation efficiencies of strains from symptomless carriers were significantly higher. The average adherence efficiency of the carriers' strains was 53 (SEM 6)% versus 35 (SEM 5)% in control strains. The average internalisation efficiency of the carriers' strains was 13.4 (SEM 4)% compared with 4.4 (SE 1.6)% in the control group. The results are in agreement with the hypothesis that, in a significant number of cases, streptococcal internalisation might contribute to eradication failure and persistent throat carriage. PMID- 10847203 TI - Comparative study of the invasiveness of Salmonella serotypes Typhimurium, Choleraesuis and Dublin for Caco-2 cells, HEp-2 cells and rabbit ileal epithelia. AB - Patterns of invasiveness of Salmonella serotypes Typhimurium, Choleraesuis and Dublin in Caco-2 cells (without centrifugation) were compared with previously published studies of the rabbit ileal invasion assay (RIIA) and (where relevant) a HEp-2 cell invasion assay. Optimal conditions for the use of Caco-2 cell monolayers in bacterial invasion assays were defined. Centrifuge-assisted attachment of bacteria to cells was not used routinely as this increased the invasiveness of known hypo-invasive strains and detachment of Caco-2 cells. Inocula with too high bacterial numbers resulted in rapid acidification of media and detachment of the monolayers. The invasiveness of Typhimurium strains TML, WAKE, WII8, LT7, SL1027 and M206 in Caco-2 cells reflected that seen in the RIIA. The invasiveness of Choleraesuis strain A50 was similar to that in the RIIA except that bacteria grown at 37 degrees C and used without storage at 4 degrees C were slightly more invasive than those grown at 37 degrees C and stored at 4 degrees C before use. Dublin strain 3246 showed no apparent temperature-regulated invasiveness in Caco-2 cells, in contrast to the results observed in the RIIA. Dublin strain 3246 did not cleave tight junctions in the Caco-2 cell monolayer as it did in rabbit ileal epithelia both in vitro and in vivo. Three TnphoA insertion LPS mutants of Typhimurium TML were uniformly hypo-invasive in both Caco-2 cells and the RIIA; in contrast, they were differentially invasive in HEp 2 cells. Three smooth TnphoA insertion mutants of Typhimurium TML (invH, invG and pagC) were hypo-invasive in both the Caco-2 and HEp-2 cell invasion assays but not in the RIIA. PMID- 10847204 TI - Accumulation of polyphosphate granules in Helicobacter pylori cells under anaerobic conditions. AB - Helicobacter pylori is known to transform to coccoid forms which might be involved in faecal-oral transmission. When the bacteria enter the intestine, they encounter anaerobiosis that is unfavourable for growth. The effect of anaerobiosis was investigated to determine whether H. pylori is viable under such conditions. H. pylori in the late logarithmic growth phase transformed from spiral to coccoid forms when transferred to and incubated anaerobically in fresh medium. Acridine orange staining indicated that the viability of coccoid forms was significantly reduced, but still measurable even at day 5 or 7 of anaerobic culture. The cells retained low but significant levels of the major sigma factor RpoD at day 5 or 7 of anaerobic culture. The cellular structures of coccoid forms contained polyphosphate granules at day 1 and even at day 7 when incubated anaerobically, whereas only a few granules were observed under micro-aerobic conditions. Poor formation of polyphosphate granules in micro-aerobic cultures correlated particularly well with lower levels of acridine orange staining. These results suggest that acridine orange-positive anaerobic coccoid forms are viable to a certain extent and that polyphosphate may support this viability. PMID- 10847205 TI - Association of gastric epithelial apoptosis with the ability of Helicobacter pylori to induce a neutrophil oxidative burst. AB - Both polymorphonuclear cell infiltration and increased epithelial apoptosis are seen in gastric mucosa in the presence of Helicobacter pylori infection. This study examined the association between bacterial ability to stimulate an oxidative burst in neutrophils and epithelial apoptosis. Biopsy specimens were obtained from 15 patients to detect apoptotic cells by the TUNEL method. H. pylori strains isolated from corresponding stomach biopsy samples were tested for the ability to stimulate an oxidative burst in human neutrophils. Neutrophils were isolated from healthy subjects without H. pylori infection and the oxidative burst was measured by flow cytometry with dichlorofluorescein diacetate. Stimulation with H. pylori increased both the percentage of activated cells and fluorescence intensity. There was a significant positive correlation between the number of epithelial apoptotic cells and fluorescence intensity. Increased neutrophil oxidative burst stimulated by H. pylori may play a role in enhanced gastric mucosal DNA damage and consequent atrophic gastritis and gastric cancer. PMID- 10847206 TI - Identification of cdtB homologues and cytolethal distending toxin activity in enterohepatic Helicobacter spp. AB - A bacterial toxin that causes progressive distension and death of Chinese hamster ovary (CHO) cells and HeLa cells, termed cytolethal distending toxin (Cdt), has been identified in several diarrhoeagenic bacteria, including Campylobacter spp. (C. jejuni and Cq coli), some pathogenic strains of Escherichia coli and Shigella spp. Genes encoding this toxin were identified as a cluster of three adjacent genes cdtA, cdtB and cdtC. Homologues of cdtB from five species of enterohepatic helicobacters (Helicobacter hepaticus, H. bilis, H. canis and two novel Helicobacter spp. isolated from mice and woodchuck, respectively) were identified by means of degenerative PCR primers, cloned and sequenced. The similarities of these partial cdtB nucleotide sequences from these Helicobacter spp. to those of cdtB genes known to be present in other bacteria were: C. jejuni, 58.3-64.8%; E. coli, 52.3-57.8%, Haemophilus ducreyi, 53.4-58.4% and Actinobacillus actinomycetemcomitans, 52.7-58.1%. Bacterial lysates from four of these helicobacters caused characteristic cytolethal distension of HeLa cells. Cdt caused cell cycle arrest at G2/M phase as measured by flow cytometry. The results demonstrated the presence of a toxin in these Helicobacter spp. belonging to the family of Cdt. PMID- 10847207 TI - Protection of BALB/c mice against experimental Helicobacter pylori infection by oral immunisation with H pylori heparan sulphate-binding proteins coupled to cholera toxin beta-subunit. AB - The presence of Helicobacter pylori in the gastroduodenal mucosae is associated with chronic active gastritis, peptic ulcers and gastric cancers such as adenocarcinoma and low-grade gastric B-cell lymphoma. In response to the presence of antibiotic-resistant strains, the use of vaccines to combat this infection has become an attractive alternative. The present study used a murine model of infection by a mouse-adapted H. pylori strain to determine whether infection in BALB/c mice can be successfully eradicated by intragastric vaccination with H. pylori heparan sulphate-binding proteins (HSBP) covalently coupled to the beta subunit of cholera toxin (CTB). It was shown that vaccination confers protection against exposure of BALB/c mice to the pathogen, as revealed by microbiological, histopathological and molecular methods. PMID- 10847208 TI - Characterisation and selection of a Lactobacillus species to re-colonise the vagina of women with recurrent bacterial vaginosis. AB - This paper reports the results of characterising and selecting a strain of Lactobacillus for potential use as a probiotic in regenerating the vaginal flora of women with recurrent episodes of bacterial vaginosis (BV). BV is a condition characterised by a depletion of vaginal lactobacilli accompanied by an overgrowth of a mixed vaginal flora of aerobic, anaerobic and micro-aerophilic species in very large numbers. BV has been associated with various gynaecological and obstetric complications and has an extremely high recurrence rate, due in part to the failure to establish a normal vaginal flora after antimicrobial therapy. A total of 60 vaginal isolates of lactobacilli was assessed for characteristics considered important for vaginal re-colonisation. The characteristics studied were the in-vitro inhibitory activity of the lactobacilli against bacterial species isolated from women with recurrent BV, acid production after growth of the lactobacilli in liquid culture, production of hydrogen peroxide (H2O2) and adhesiveness of the lactobacilli to exfoliated vaginal epithelial cells (VEC). Four strains of lactobacilli, L. acidophilus (61701 and 61880), L. crispatus (55730) and L. delbrueckii subsp. delbrueckii (65407), demonstrated the greatest inhibitory activity against the BV-associated bacterial species. Two of these isolates (55730 and 61880) produced H2O2. All four isolates produced a highly acidic environment after growth in liquid medium (pH <4). Only one of these (strain 61701) was strongly adherent to VEC (>100 bacteria/VEC). A further isolate (L. acidophilus 48101) did not demonstrate maximum inhibitory activity against BV-associated bacteria, but was found to be a strong producer of H2O2 and was also highly adherent to VEC. Isolates 61701 and 48101 could be candidates for use as probiotics for vaginal re-colonisation. PMID- 10847209 TI - Efficacy of influenza vaccine in the elderly in welfare nursing homes: reduction in risks of mortality and morbidity during an influenza A (H3N2) epidemic. AB - The effect of influenza vaccination on the occurrence and severity of influenza virus infection in a population residing in nursing homes for the elderly was studied during an influenza A (H3N2) epidemic in Japan. Of 22,462 individuals living in 301 welfare nursing homes, 10,739 received either one dose (2027 subjects) or two doses (8712 subjects) of inactivated, subunit trivalent influenza vaccine. During the period Nov. 1998 to March 1999, there were 950 cases of influenza infection diagnosed clinically, with virus isolation or serology. There were statistically significantly fewer cases of influenza, hospital admissions due to severe infection and deaths due to influenza in the vaccinated cohort (256 cases, 32 hospital admissions, 1 death) than in the unvaccinated controls (694 cases, 150 hospital admissions, 5 deaths; reduction rates 59.8%, 76.9% and 79.1% respectively). Vaccination was almost equally effective in those who received one dose of vaccine and those who received two doses. No serious adverse reactions to vaccination were recorded. Thus influenza vaccination is safe and effective in this population, and should be an integral part of the routine care of persons aged > or =65 years residing in nursing homes. PMID- 10847210 TI - Prolonged perturbations of tumour necrosis factor-alpha and interferon-gamma in mice inoculated with Clostridium piliforme. AB - Clostridium piliforme is an obligate intracellular bacterium that causes enterohepatic disease in many animal species. C. piliforme infections are commonly subclinical in laboratory rats and mice, and little is known about host regulation of disease or of the effects of C. piliforme infections on investigations that use subclinically infected animals. To assess host regulation of subclinical C. piliforme infections and the effects of those infections on laboratory mice, the expression of the pro-inflammatory cytokines tumour necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) was evaluated at 0, 1, 3, 7, 14 and 28 days after inoculation with C. piliforme. Subclinical infection was induced in weanling C. piliforme-susceptible DBA/2 or -resistant C57BL/6 mice with either a toxic or a non-toxic C. piliforme strain. Hepatic lesions and bacteria were demonstrated histologically in both mouse strains for 14 days after inoculation with the toxigenic bacterial strain, but were never demonstrated histologically following inoculation with the non-toxigenic strain. Hepatic TNF alpha and IFN-gamma mRNA and serum protein levels were similarly elevated in both mouse strains 1 day after inoculation with both C. piliforme strains, as evaluated by reverse transcription PCR and enzyme-linked immunosorbent assays, respectively. Elevation of IFN-gamma persisted for 14 days after inoculation; TNF alpha remained elevated at 28 days after inoculation. PMID- 10847211 TI - Genotypic variations of Shiga toxin-converting phages from enterohaemorrhagic Escherichia coli O157: H7 isolates. AB - Pulsed-field gel electrophoresis (PFGE) analysis revealed that enterohaemorrhagic Escherichia coli (EHEC) O157:H7 strains had considerable variations in their genomes. This study investigated whether or not the molecular profile of Shiga toxin (Stx) 1- and Stx2-converting phages isolated from EHEC O157:H7 strains, derived from various sources in the USA and Japan, corresponded to the variations of host strains' genotypes as determined by PFGE. A total of 51 Stx-converting phages including 12 Stx1-converting phages and 37 Stx2-converting phages was isolated from seven USA isolates and 20 Japanese isolates. The average Dice coefficient values showed 44% similarity between phage DNAs in Stx2-converting phages digested with SmaI and 55% in Stx1-converting phages digested with HindIII, indicating considerable variation among phage DNA. In particular, restriction fragment length polymorphism (RFLP) patterns of Stx2-converting phage DNA varied according to the PFGE type of their host strain, which suggests that the phage genomes have altered their genotypic characteristics with those of host genomes. However, there are several exceptions: the RFLP patterns of some Stx2 converting phages were quite similar irrespective of the different genotypes of the host strains, indicating that horizontal transfer of Stx2-converting phage may also occur under some circumstances. PMID- 10847212 TI - Molecular genotyping of Candida species with special respect to Candida (Torulopsis) glabrata strains by arbitrarily primed PCR. AB - A set of 46 epidemiologically related or unrelated Candida (Torulopsis) glabrata isolates from four different medical centres in Germany and Hungary, and the type strain of this species, were genetically typed by arbitrarily primed PCR (AP PCR). The resulting band patterns of C. glabrata strains were compared with those of other species of the genus Candida including C. albicans, C. guilliermondii, C. kefyr, C. parapsilosis, C. tropicalis and C. krusei. After preliminary trials of various reaction parameters and control experiments to test the reproducibility of this method, it was found that consistently reproducible amplification patterns were obtained only when rigorously optimised and standardised reaction conditions were employed. Discriminatory abilities were studied with 29 generated 10-mer oligonucleotides of different G+C content. Typing of clinical isolates with the optimised AP-PCR protocol was then performed with the primer 50-1, with a G+C content of 50%. Sufficiently discriminatory polymorphisms were observed among the band patterns of the Candida species included. The gel electrophoresis patterns of each species showed an adequate similarity. Variations in minor bands were characteristic for comparison at the isolate level. Only three AP-PCR genotypes were identified among the clinical isolates of C. glabrata tested. Two of these genotypes were closely related and appeared to be widespread within German and Hungarian isolates. The third genotype of C. glabrata showed a distinct band pattern. With optimised, validated and standardised assay conditions, the feasibility, sensitivity and rapidity of AP-PCR may offer a discriminatory method for genotyping of yeasts in epidemiological studies, as well as in the control of nosocomial infections. PMID- 10847213 TI - Drug information systems, early warning, and new drug trends: can drug monitoring systems become more sensitive to emerging trends in drug consumption? AB - Drug Information Systems (DIS) are called upon to provide an early warning of emerging trends in drug use. However, little theoretical attention is directed toward exploring conceptual issues in this area. In this paper a typology of existing DIS is offered. Among the features that distinguish DIS are their structure (human network or organization systems) and the range of information sources used. Indicators of drug use can be placed on a continuum of sensitivity ranging from leading edge indicators to lagged indicators. Sensitivity implies volatility as sensitive indicators also react to fluctuations that do not become trends. DIS conventionally are largely reliant upon lagged indicators. What is required are DIS that combine a critical information processing function with the ongoing systematic collection of data from a range of data sources. PMID- 10847214 TI - Response effects due to bystander presence in CASI and paper-and-pencil surveys of drug use and alcohol use. AB - In this study we investigated the influence of bystanders on self-administered interviews asking about the use of alcohol and illicit drugs. Interview participants were adolescents and young adults living in urban and suburban areas of the United States. Participants were assigned randomly to either a computerized or a paper-and-pencil self-administered interview. Results show that the impact of bystanders during the interview varies according to the identity of the bystander, age of the person interviewed, and the mode of interview. When a parent was present during the interview, survey participants were less likely to report the use of alcohol and marijuana. The influence of parents was stronger for adolescents than for young adults. The use of computer-assisted self administered interviewing, compared to interviews with paper-and-pencil forms, reduced the effects due to the presence of parents during the interview. The presence of siblings during the interview had a small, negative effect on reports of using alcohol or illicit drugs. Among married or cohabiting respondents, the presence of the husband, wife, or live-in partner had no influence on reports of alcohol use or drug use. PMID- 10847215 TI - The impact of T-ACASI interviewing on reported drug use among men who have sex with men. AB - Measurements of drug use and other illicit or stigmatized behaviors are subject to nontrivial underreporting biases. During in-person surveys, respondents are more likely to report such behaviors when interviewed using techniques that maximize interviewee privacy, e.g., use of paper SAQs and audio-CASI rather than questioning by human interviewers. Until recently, respondents in telephone surveys could not be offered similar privacy. A new technology, telephone audio computer-assisted self-interviewing (T-ACASI) overcomes this limitation of telephone surveys by allowing respondents to respond to a computer. A randomized experimental test of T-ACASI was embedded in the Urban Men's Health Study (UMHS). UMHS surveyed a probability sample of 2,881 men from four United States cities and who reported having sex with men. Respondents interviewed using T-ACASI reported a higher prevalence of drug use and drug-related behaviors than respondents interviewed by human interviewers. However, survey respondents were more likely to break off an interview when the interview was conducted by a T ACASI computer rather than by a human interviewer. PMID- 10847216 TI - Survey estimates of drug-use trends in urban communities: general principles and cautionary examples. AB - Surveys to depict substance abuse rates and monitor trends in specific areas have become increasingly important policy tools. Yet, as illustrated by two national multiwave surveys, using small sample survey data and making longitudinal comparisons is fraught with interpretative problems. In the case of the metropolitan area "oversample" of the National Household Survey on Drug Abuse, for example, interpreting apparent declines in drug use has to take account of the devastating effects of Hurricane Andrew in the Miami Metropolitan area. In the case of a 41-community survey sponsored by the Robert Wood Johnson Foundation to evaluate substance abuse prevention, the difficulty is how to interpret small differences in drug use, which seem to follow no reasonable pattern with respect to treatment or comparison sites. Inferences from such surveys are confounded with statistical anomalies and unforeseen events. They are limited by the sample size. In part, the solution to these problems is to use other survey and nonsurvey data to validate their conclusions and to note their limitations. PMID- 10847217 TI - Age, period, and cohort effects in marijuana and alcohol incidence: United States females and males, 1961-1990. AB - Using multiple classification models applied to self-report data on initiation of drug use from nine National Household Surveys on Drug Abuse conducted between 1982 and 1995, this paper shows that the directions of change in period and cohort effects were similar for marijuana and alcohol and for males and females. Period effects--indicative and societal tolerance or support for drug use during 5-year periods between 1961 and 1990--declined between the early 1970s and late 1980s, while cohort effects--indicative of early experiences of birth cohorts favoring drug use--increased. One interpretation is that trends in incidence were determined by two opposing vectors of social forces: Beginning in the 1970s, changes in social policies, values, and drug markets--as reflected in period effects--increasingly acted to reduce incidence, while changes in conditions of childhood socialization--as reflected in cohort effects--increasingly facilitated or encouraged incidence. Especially for marijuana, the increase in cohort effects is larger among females, which gives rise to gender convergence--approximately equal male and female incidence rates for both drugs--by the late 1980s. An innovative method of the paper is the adjustment of incidence rates for reporting bias. PMID- 10847218 TI - Trends in substance use among US military personnel: the impact of changing demographic composition. AB - This paper examines the impact of the military population's changing demographic composition on observed changes in substance use by military personnel. Cross sectional data are drawn from the six Department of Defense Worldwide Surveys of Substance Abuse and Health Behaviors Among Military Personnel conducted in 1980, 1982, 1985, 1988, 1992, and 1995. Using the method of direct standardization to adjust for changes in demographic composition, trends in substance use are examined. Changes in heavy alcohol use were more affected by demographic changes in the military population than were changes in illicit drug use or cigarette use. Findings indicate that changes in demographic composition may need to be controlled when assessing trends from cross-sectional surveys. PMID- 10847219 TI - Nonresponse and selection bias in treatment follow-up studies. AB - Large-scale studies of addiction treatment employ two stages: select providers, then select patients to follow-up. Nonresponse due to noncooperation of providers and problems of locating and recruiting patients may bias the results. We review selection and attrition biases in previous work and in four major United States treatment studies in the 1990s: DATOS (N = 10,100 clients, 96 units), NTIES (N = 6,593/71), SROS (N = 3,047/99), and CALDATA (N = 3,045/86). We develop a standard approach, break down sampling and attrition rates, and discuss differences in client, program, and methodology factors. We conclude with some methodological recommendations for future follow-up studies of addiction treatment. PMID- 10847221 TI - The use of the case-crossover design in studying illicit drug use. AB - The case-crossover design was developed to study time-varying exposures that cause transient excess risk of acute health events. It is a variant of case control and subject-as-own-control research designs, involving use of information about exposure history of each case to estimate the transient effect. This kind of self-control design can help to reduce sampling bias otherwise introduced in the selection of controls, as well as confounding bias that might be derived from enduring individual characteristics, especially personality traits and other long standing inherited or acquired vulnerabilities. When the subject is used as his or her own control, these personal vulnerabilities are matched. In this paper we discuss strengths and weaknesses of the case-crossover design and suggest applications of the case-crossover design in epidemiologic studies on suspected hazards of illicit drug use, and in studies of drug use and co-occurring psychiatric disturbances. We conclude that the case-crossover design can play a useful role, but it discloses a need to secure fine-grained measurements in epidemiologic research on psychiatric comorbidity. As explained in the paper, we also believe the case-crossover method may be of use to criminologists who study the drugs-crime nexus, to services researchers and clinicians who seek to understand treatment entry and compliance behavior, and to etiologists interested in polydrug use. PMID- 10847220 TI - Inconsistencies over time in young adolescents' self-reports of substance use and sexual intercourse. AB - When surveyed twice within a 6-month period, to what extent do young adolescents give inconsistent answers to questions about sexual behavior and substance use (cigarettes, alcohol, marijuana, inhalants, cocaine)? Data were collected from 1,575 urban African American and Hispanic students during fall and spring of 7th grade. For each behavior examined, less than 2% of the sample gave inconsistent answers within a survey and less than 7.5% did so over time. Retraction of baseline answers at follow-up was greater for rarer and more socially undesirable behaviors (e.g., cocaine versus cigarette use). Over-time inconsistencies were associated with lower reading comprehension and an overall reduction in risk behaviors at follow-up. PMID- 10847222 TI - Use of alternating logistic regression in studies of drug-use clustering. AB - This article describes the alternating logistic regression (ALR) method, and places this method in the context of other statistical approaches to the analysis of complex survey data, including the conditional form of logistic regression with matching on neighborhood characteristics. Unlike conditional logistic regression, the ALR method provides for an explicit estimation of the magnitude of clustering of drug use within neighborhoods and within subgroups of the neighborhood defined by male-female or age indicators, with and without covariate adjustments. The application of these ALR methods is illustrated with estimates for the magnitude of clustering of daily marijuana use and weekly marijuana use within neighborhoods of the United States, based on data from the National Household Survey on Drug Abuse samples from 1990 through 1996. PMID- 10847223 TI - Racial/ethnic and educational differences in the estimated odds of recent nitrite use among adult household residents in the United States: an illustration of matching and conditional logistic regression. AB - This article compares estimates of the relative odds of nitrite use obtained from weighted unconditional logistic regression with estimates obtained from conditional logistic regression after post-stratification and matching of cases with controls by neighborhood of residence. We illustrate these methods by comparing the odds associated with nitrite use among adults of four racial/ethnic groups, with and without a high school education. We used aggregated data from the 1994-B through 1996 National Household Survey on Drug Abuse (NHSDA). Difference between the methods and implications for analysis and inference are discussed. PMID- 10847224 TI - The reproductive endocrine consequences of anorexia nervosa. PMID- 10847225 TI - Fertility following radical, conservative-surgical or medical treatment for tubal pregnancy: a population-based study. AB - OBJECTIVE: To investigate the factors influencing the choice of treatment for ectopic pregnancy and to compare the subsequent fertility rates of radical, conservative-surgical or medical treatments. DESIGN: Population-based study of 835 ectopic pregnancies registered between 1992 and 1996. SETTING: Auvergne Ectopic Pregnancy Register (France). SAMPLE: Four hundred and seventy-six women with tubal ectopic pregnancy who were not using contraception at the time of conception. Subsequent fertility was studied for the 291 women who attempted to conceive again. METHODS: Comparison of reproductive outcomes according to ectopic pregnancy treatment. Cumulative fertility curves were calculated by the Kaplan Meier estimator and compared by log rank test for univariate analysis and by Cox regression to take into account confounding variables. MAIN OUTCOME MEASURE: Recurrence and fertility rates after ectopic pregnancy. RESULTS: The first treatment given was 'radical' for 178 women (37%), 'conservative-surgical' for 262 (55%), and 'medical' for 35 (8%). The treatment failed in 1% for radical treatment, 5% for conservative-surgical treatment, and 36% for medical treatment. The two-year cumulative rate of recurrence was 27% with no significant difference between treatments. For women with previous infertility factors (in particular diseased contralateral tube), the treatments differed significantly, with the rate of intrauterine pregnancy lower for radical treatment and higher for medical treatment than for conservative-surgical treatment. For women with no infertility factor, there was no significant difference between treatments. CONCLUSIONS: These results should be confirmed in a controlled trial. The results of this study provide the elements necessary to plan such a trial. PMID- 10847226 TI - A study to evaluate serum and urinary hormone levels following short and long term administration of two regimens of progesterone cream in postmenopausal women. AB - OBJECTIVE: To determine the pharmacokinetics of a progesterone cream following short and long term dermal administration. DESIGN: Single-centre, randomised, multiple-dose, open-label study. SETTING: Reproductive Medicine Trust, London. POPULATION: Twenty-four healthy postmenopausal women aged between 40 and 65 years were recruited through an advertisement in a local newspaper. METHODS: The women were randomly allocated to progesterone cream 40 mg daily or 20 mg, twice daily, for 42 days. MAIN OUTCOME MEASURES: The concentration of progesterone in the serum was measured on days 1 and 42 before the morning dose, and at 2, 4, 6, 12 and 24 hours after the morning dose. Serum follicle stimulating hormone, oestradiol, testosterone and urinary pregnanediol-3-glucuronide were also measured on days 1 and 42. RESULTS: Three subjects dropped out before using the cream and two more dropped out after the first treatment leaving a reportable sample of 19 women. There was a rise in the mean progesterone concentration at each sampling time between days 1 and 42. There was evidence of a rise in pregnanediol-3-glucuronide over the course of the study. There was no change in follicle stimulating hormone, oestradiol or testosterone. There was no difference between the two regimens. CONCLUSIONS: Transdermal progesterone (40 mg) per day for 42 days causes a small increase in serum progesterone concentration, although there is wide variation. Whether such levels are of clinical benefit remains to be seen. PMID- 10847227 TI - Should tissue from pregnancy termination and uterine evacuation routinely be examined histologically? AB - OBJECTIVE: To assess the value of routine histological examination of tissue samples collected at termination of pregnancy in the first trimester and emergency surgical uterine evacuation. SETTING: The gynaecological department of a teaching hospital. DESIGN: Prospective study of women attending the gynaecological department in a 12-month period. PARTICIPANTS: All women undergoing a therapeutic first trimester medical or surgical abortion or an emergency surgical evacuation of a failed pregnancy, suspected incomplete spontaneous miscarriage or incomplete induced abortion. MAIN OUTCOME: Association of pre-operative clinical diagnosis and the post-operative histological result. RESULTS: Of 1,576 women studied, the histological report confirmed that products of conception were obtained in 1,465 (93%); in two women (0.13%) molar changes were reported confirming the preoperative diagnosis by ultrasound. Products of conception were not confirmed in the tissue specimens in 0.5% medical terminations, 5% surgical terminations, 10% evacuations following a previous evacuation, 12% evacuations for a failed pregnancy, and 19% evacuations for an incomplete miscarriage. In 87 women (6%), decidua was reported; two of these women had undergone an evacuation for an ultrasound diagnosis of spontaneous miscarriage, but in both a tubal ectopic pregnancy was subsequently diagnosed. CONCLUSION: There did not appear to be any obvious benefit from routine histological examination of tissue removed at termination of pregnancy or emergency uterine evacuation. The histological result was sometimes not consistent with the pre-operative diagnosis and may result in unnecessary further investigation and treatment unless due consideration is given to the clinical presentation. PMID- 10847228 TI - Social and behavioural influences on the uptake of hormone replacement therapy among younger women. AB - OBJECTIVE: To describe the social and behavioural influences on the uptake of hormone replacement therapy before the age of 50. DESIGN: Nationally representative birth cohort study with detailed hormone replacement therapy histories and prospective data on health, social and behavioural factors collected throughout life. SETTING: England, Scotland and Wales. POPULATION: General population sample of 1,572 women followed to the age of 50 years. MAIN OUTCOME MEASURE: Age at first hormone replacement therapy use RESULTS: By the age of 50 years, 45% of women had tried hormone replacement therapy and one third were current users. Over two-fifths of users had tried more than one preparation and over one quarter had episodic use. For the vast majority, prescribing conformed to current guidelines. More educated women took hormone replacement therapy for long term prevention compared with their less educated peers. Hysterectomy increased the chances of taking hormone replacement therapy, particularly where an oophorectomy had also been performed, and was associated with longer and more continuous use. Results of a Cox's proportional hazards model showed that the age at which first hormone replacement therapy is used by women who have not had a hysterectomy was influenced by previous contact with health services for menstrual disorders, previous use of oral contraception and cigarettes, past reporting of health problems and low social class. The relation between smoking and low social class with early use of hormone replacement therapy may be due to their association with early menopause. CONCLUSIONS: The trend over the last two decades towards greater use of hormone replacement therapy has continued unabated for younger women. So far, hormone replacement therapy users in this generation have had less healthy lifestyles and social advantages than nonusers, in contrast to many older mainly American studies based on earlier generations. This may have long term implications for health and health care as the postwar baby boom generation ages. PMID- 10847229 TI - A population study of birthweight and the risk of caesarean section: Scotland 1980-1996. AB - OBJECTIVES: 1. To describe the relation between birthweight and risk of emergency caesarean section at term; 2. to determine whether the relation between birthweight and caesarean section differed between male and female babies; and 3. to determine what proportion of the increased rates of caesarean section could be related to greater birthweights. DESIGN: Retrospective population study. SETTING: Data collected from Scottish Morbidity Record 1980 to 1996. POPULATION: All first singleton deliveries by emergency caesarean section and non-elective vaginal birth of live babies at 40 weeks of gestation (n = 120,854). MAIN OUTCOME MEASURE: Delivery by emergency caesarean section. RESULTS: There was a U-shaped relation between birthweight and the risk of caesarean section, with the lowest risk associated with weights in the range 3,000-3,500 g. Overall, male, were more likely to be delivered by caesarean section (relative risk = 1.2, 95% CI 1.2 1.3). The association between male sex and increased risk of caesarean section persisted after adjusting for birthweight, but only males weighing < 4,000 g were at increased risk of caesarean section compared with similarly sized females. Between 1980 and 1996, there were linear increases in the rate of caesarean section (from 7.1% to 10.7%, r2 = 0.8, P < 0.001) and median birthweight (from 3,360 g in 1980 to 3,420 g in 1996, r2 = 0.8, P < 0.001). The population attributable fraction of caesarean sections related to year of delivery 1981-1996 was not significantly altered by adjusting for birthweight (22.3% vs 21.6%). CONCLUSIONS: There is no evidence to suggest that increasing birthweights have contributed to increasing rates of caesarean section in Scotland between 1980 and 1996 among singleton first births non-electively delivered at 40 weeks of gestation. PMID- 10847230 TI - The effect of smoking on pre-eclampsia in twin pregnancy. AB - OBJECTIVE: To assess the effect of smoking on the incidence of pre-eclampsia and on perinatal outcome in twin pregnancy. DESIGN: Retrospective study using Aberdeen Maternity and Neonatal Databank. SETTING: Grampian, Orkney and Shetland. POPULATION: All 1,575 twin pregnancies delivered in the years 1969-1971 and 1976 1997 (when smoking data were available). METHODS: In all twin pregnancies the effects of smoking on pre-eclampsia were analysed by parity and gestation at delivery. The effect of smoking upon late miscarriage and perinatal outcome was analysed without subdivision. MAIN OUTCOME MEASURES: Pre-eclampsia and perinatal outcome. RESULTS: Primiparae had significantly higher rates of pre-eclampsia than multiparae and were delivered significantly earlier. The incidence of pre eclampsia in smokers was significantly lower only in multiparae. Length of gestation was significantly shorter in multiparous smokers. The effect of smoking on pre-eclampsia appeared to be direct in multiparae but possibly indirect in primiparae (by causing earlier delivery). Smokers had a higher late miscarriage rate than nonsmokers. CONCLUSIONS: In twin pregnancy the apparent protective effect of smoking against pre-eclampsia is significant only in multiparae, suggesting that in primiparae the smoking effect is overwhelmed by the other reasons for the development of pre-eclampsia. Smoking in twin pregnancy is not recommended due to the worse fetal outcome rates. PMID- 10847231 TI - The relationship between intrauterine growth restriction and preterm delivery: an empirical approach using data from a European case-control study. AB - OBJECTIVE: To test whether being small for gestational age, defined as having a birthweight less than the 10th centile of intrauterine growth references, is a risk factor for preterm delivery for singleton live births. DESIGN: A case control study. SETTING: Maternity hospitals in 16 European countries. SAMPLE: Four thousand and seven hundred preterm infants between 22 and 36 completed weeks of gestation and 6,460 control infants between 37 and 40 weeks of gestation. METHODS: Newborn babies are identified as being small for gestational age using customized reference standards derived from models of fetal growth. The impact of being small for gestational age on preterm delivery is estimated using logistic regression. MAIN OUTCOME MEASURE: Spontaneous or induced preterm delivery. RESULTS: Being small for gestational age is significantly associated with preterm birth, although the magnitude of this association differs greatly by type of delivery and gestational age. Over 40% of induced preterm births for reasons other than the premature rupture of membranes are small for gestational age compared with 10.7% of control infants (OR 6.41). For spontaneous or premature rupture of membranes related preterm births, the association is also significant, but weaker (OR 1.51). The relationship between growth restriction and preterm delivery is strongest for preterm births before 34 weeks of gestation. CONCLUSIONS: These findings highlight the phenomenon of abnormal fetal growth in all premature infants and, in particular, infants delivered by medical decision for reasons other than premature rupture of membranes. The observed association between being small for gestational age and preterm delivery among spontaneous preterm births merits further attention because the causal mechanisms are not well understood. PMID- 10847232 TI - Nifedipine or prazosin as a second agent to control early severe hypertension in pregnancy: a randomised controlled trial. AB - OBJECTIVE: To determine whether nifedipine or prazosin is the more appropriate second-line antihypertensive agent in pregnancy. DESIGN: Randomised controlled trial. SETTING: Tygerberg Hospital, a tertiary referral centre. POPULATION: Women with early, severe pre-eclampsia or hypertension in pregnancy, whose blood pressure could not be adequately controlled by methyldopa 2 g/day, but were otherwise stable. METHODS: Nifedipine or prazosin were given and increased as necessary in a stepwise fashion. Once the maximum dose was reached, the other drug was added in a crossover pattern. Failure to control blood pressure, or the onset of maternal/fetal complications were indications for delivery. Patients reaching a minimum gestation of 34 weeks without complications were delivered electively. MAIN OUTCOME MEASURES: Antenatal days gained; major maternal complications and perinatal survival. RESULTS: Days gained on the second antihypertensive agent did not differ significantly (P = 0.9), while more days were gained using nifedipine as the crossover 'third agent' (P = 0.01). In the nifedipine group better renal function was recorded, but more cases with isolated low platelet counts occurred. More cases of pulmonary oedema as well as more nonviable mid-trimester and third trimester intrauterine deaths occurred in the prazosin group. CONCLUSION: Nifedipine and prazosin as second agents allowed comparable amounts of time to be gained, although this changed when used as crossover third-line agents. The efficacy and safety of nifedipine in this study are consistent with the results of other studies. A greater number of intrauterine deaths occurred in the prazosin group. PMID- 10847233 TI - Detection of fetal Rhesus D and sex using fetal DNA from maternal plasma by multiplex polymerase chain reaction. AB - OBJECTIVE: To test the sensitivity, specificity and reproducibility using fetal DNA obtained from plasma of pregnant women by polymerase chain reaction for the simultaneous detection of both fetal sex and Rhesus D genotype. METHOD: Blood samples were obtained from 22 Rhesus D negative pregnant women about to undergo an invasive procedure. DNA was extracted from the plasma fraction and analysed by a multiplex nested polymerase chain reaction using Y chromosome-and Rhesus D specific primers. The results of this experimental procedure were compared with those obtained from the analysis performed on material gained by the invasive procedure. RESULTS: The sensitivity of the plasma polymerase chain reaction-based method was surprisingly high, with both fetal genotypes being correctly determined in almost 100% of the cases examined. In only one instance was a false positive result for the detection of Rhesus D recorded, which on subsequent analysis was negative. CONCLUSIONS: The ease and rapidity with which the plasma polymerase chain reaction-based method can be performed makes it a promising method for the analysis of multiple fetal loci, such as Rhesus D and sex. PMID- 10847234 TI - The transplacental transfer of the macrolide antibiotics erythromycin, roxithromycin and azithromycin. AB - OBJECTIVE: To investigate the transplacental transfer of the macrolide antibiotics erythromycin, roxithromycin and azithromycin. METHODS: Twenty-one term placentas were obtained with maternal consent immediately after delivery and a two-hour nonrecirculating perfusion of a single placental cotyledon was performed. Erythromycin (2 microg/mL), roxithromycin (2 microg/mL) and azithromycin (0.3 microg/mL) were infused to the maternal inflow at a constant rate, with antipyrine as a reference compound, and their appearance in the fetal circulation was followed. Drug concentrations were measured by high performance liquid chromatography for 120 min. RESULTS: The mean transplacental transfers (TPT(SS)) for erythromycin, roxithromycin and azithromycin were 3.0%, 4.3% and 2.6%, respectively, calculated as the ratio between the steady state concentrations in fetal venous and maternal arterial sides. Similar results were obtained when the TPT was calculated as the absolute amount of drug transferred across the placenta during 2-hour perfusion (TPT(A)). No significant differences were found among the three macrolides in TPT(SS) (P = 0.39) or TPT(A) (P = 0.35). The TPT(SS) of erythromycin, roxithromycin and azithromycin were 41%, 35% and 32% of the freely diffusable reference compound antipyrine, respectively. Steady state was reached in 60 minutes in each perfusion indicating sufficient perfusion time. CONCLUSION: The limited transplacental transfer of erythromycin, roxithromycin and azithromycin suggests compromised efficacy in the treatment of fetal infections. On the other hand, the placenta seems to produce an effective barrier reducing the fetal exposure when these three macrolides are used to treat maternal infections. PMID- 10847235 TI - Dyslipoproteinaemia in postmenopausal women with a history of eclampsia. AB - OBJECTIVE: To test the hypothesis that postmenopausal women with a history of eclampsia manifest a more high risk lipid profile than postmenopausal women with a history of normal pregnancy. SETTING: The Department of Obstetrics and Gynaecology, National University Hospital, Reykjavik, Iceland, and the Magee Womens Research Institute, Pittsburgh, Pennsylvania, USA. PARTICIPANTS: Thirty Icelandic women with a history of eclampsia, aged between 50 and 67 years at the time of re-examination (cases) were individually matched for current age, and for age and parity at index pregnancy, to 30 unrelated Icelandic women with a history of normal pregnancy (controls). METHODS: The participating women completed a health and family history questionnaire and underwent a physical examination. Fasting plasma low density lipoprotein diameter, serum lipids, insulin, and glucose were measured. RESULTS: Mean low density lipoprotein size was significantly smaller and apolipoprotein B concentration was higher in women with prior eclampsia. The percentage of cases receiving blood pressure medication (33%) was significantly greater than controls (6.7%). Thirteen cases had had hypertensive complications in at least one other pregnancy (recurrent subgroup); postmenopausally, these women displayed significantly increased diastolic blood pressures, smaller-sized low density lipoprotein, increased apolipoprotein B, decreased high density lipoprotein2 (HDL2) cholesterol, and increased total cholesterol: HDL cholesterol ratio compared with their controls. Fourteen cases were normotensive in all other pregnancies (nonrecurrent); these showed no differences from their controls. CONCLUSIONS: Dyslipoproteinaemia is more prevalent among postmenopausal women with prior eclampsia, especially with recurrent hypertension in pregnancy, than in postmenopausal women with prior normal pregnancies. PMID- 10847236 TI - Hyperhomocysteinemia and other thrombotic risk factors in women with placental vasculopathy. AB - OBJECTIVE: To investigate coagulation inhibitors and abnormalities of the homocysteine metabolism, which are related to an increased thrombotic risk, as risk factors for placental vasculopathy. DESIGN: A case-control study comparing nonpregnant women with an obstetric history of placental vasculopathy (study group) with nonpregnant women (control group) matched for age and occupation. SETTING: Obstetric outpatient clinic in the University Hospital Nijmegen. SAMPLE: One hundred and one women in the study group and 92 women in a control group. METHODS: Determinations in blood samples of homocysteine concentrations; the occurrence of 677 C-->T mutation in the methylenetetrahydrofolate reductase (MTHFR) gene; protein C activities; activated protein C resistance ratios; concentrations of free protein S antigen; antithrombin III activities; and the occurrence of factor V Leiden mutation. RESULTS: Increased risk for placental vasculopathy was found in the study group with elevated homocysteine (odds ratio 2.28, 95% CI 1.18-4.39), MTHFR mutation (odds ratio 3.29, 95% CI 1.03-10.5), decreased activated protein C resistance ratio (odds ratio 2.46, 95% CI 1.06 5.72) and protein C (odds ratio 2.01, 95% CI 1.11-3.65). Any combination of two risk factors in the same individual resulted in a 3.40 (95% CI 1.80-6.42) higher relative risk for placental vasculopathy; combinations of three risk factors in a 6.83 (95% CI 1.52-30.7) higher risk. CONCLUSIONS: The thrombotic risk factors decreased levels of activated protein C resistance ratios and protein C, elevated homocysteine and the MTHFR 677 C-->T mutation are likely risk factors for placental vasculopathy. Combinations of these risk factors in one individual resulted in synergistic increase of risk. PMID- 10847237 TI - Nifedipine administered in pregnancy: effect on the development of children at 18 months. AB - The safety of use of the calcium channel blocker nifedipine in pregnancy as it affects child development has not been well evaluated. We report the results, with regard to the safety for children of use of nifedipine in pregnancy, on children followed up at 18 months of age born from women recruited in a study comparing routine treatment with nifedipine compared with no treatment. PMID- 10847238 TI - Raised maternal serum inhibin A concentration at 10 to 14 weeks of gestation is associated with pre-eclampsia. AB - Maternal serum inhibin A and free beta human chorionic gonadotrophin (beta-hCG) were measured in 759 chromosomally normal, pregnant women at 10-14 weeks of gestation. There were nine who subsequently developed pre-eclampsia and in these women the maternal serum inhibin A concentration was significantly higher than in the normotensive controls. PMID- 10847239 TI - Maternal thyroid disease: a risk factor for newborn encephalopathy in term infants. AB - Two previously published studies of term newborn encephalopathy showed that maternal thyroid disease to be a risk factor. From these studies we identified 13 case and three control mothers with thyroid disease and investigated them further. The majority of affected case mothers had idiopathic or autoimmune hypothyroidism. Compared with control mothers, case mothers had fewer thyroid function tests in pregnancy, were more likely to remain on the same dose of medication throughout pregnancy and to have experienced other pregnancy complications. The association between maternal thyroid disease and encephalopathy may be the result of a series of different causal pathways, some of which are suggested by our data. PMID- 10847240 TI - Placenta increta: evaluation of radiological investigations and therapeutic options of conservative management. PMID- 10847241 TI - Rupture of the uterus after 800 micrograms misoprostol given vaginally for termination of pregnancy. PMID- 10847242 TI - Vomiting and hyponatraemia in pregnancy. PMID- 10847243 TI - Pregnancy-dependent Cushing's syndrome in three pregnancies. PMID- 10847244 TI - Leptin concentrations in maternal serum and cord blood: relationship to maternal anthropometry and fetal growth. PMID- 10847245 TI - Reducing perinatal and maternal mortality in the world: major challenges. PMID- 10847246 TI - Is an ultrasound assessment of gestational age at the first antenatal visit of value? A randomised clinical trial. PMID- 10847247 TI - The patient's response to medical futility. PMID- 10847248 TI - Elder abuse and neglect. Autopsy Committee of the College of American Pathologists. PMID- 10847249 TI - American Thyroid Association guidelines for detection of thyroid dysfunction. AB - OBJECTIVE: To define the optimal approach to identify patients with thyroid dysfunction. PARTICIPANTS: The 8-member Standards of Care Committee of the American Thyroid Association prepared a draft, which was reviewed by the association's 780 members, 50 of whom responded with suggested revisions. EVIDENCE: Relevant published studies were identified through MEDLINE and the association membership's personal resources. CONSENSUS PROCESS: Consensus was reached at group meetings. The first draft was prepared by a single author (P.W.L.) after group discussion. Suggested revisions were incorporated after consideration by the committee. CONCLUSIONS: The American Thyroid Association recommends that adults be screened for thyroid dysfunction by measurement of the serum thyrotropin concentration, beginning at age 35 years and every 5 years thereafter. The indication for screening is particularly compelling in women, but it can also be justified in men as a relatively cost-effective measure in the context of the periodic health examination. Individuals with symptoms and signs potentially attributable to thyroid dysfunction and those with risk factors for its development may require more frequent serum thyrotropin testing. PMID- 10847250 TI - Update on antiplatelet therapy for stroke prevention. AB - The high rates of mortality and long-term disability associated with ischemic stroke, coupled with its prevalence, necessitate good, long-term preventive strategies. Risk-factor management is effective for individuals with preclinical and clinical cerebrovascular disease. Patients suffering from a transient ischemic attack or stroke are particularly vulnerable to subsequent stroke. Most of these individuals are candidates for antiplatelet treatment to prevent a recurrence. Available antiplatelet therapies include aspirin, ticlopidine, and clopidogrel. The combination of low-dose aspirin plus extended-release dipyridamole has been shown to offer safe, effective antiplatelet therapy for appropriate patients. In the second European Stroke Prevention Study, the combination was found to be significantly more effective than either drug alone, at the cost of relatively few treatment-related adverse effects. This combination is currently recommended as one of the first-line treatments for stroke prevention after first transient ischemic attack or stroke. PMID- 10847251 TI - Hypertension and diabetes: new therapeutic options. AB - The treatment of high-risk hypertensive patients with diabetes presents clinicians with challenges and opportunities. The coexistence of hypertension and diabetes dramatically and synergistically increases the risk of microvascular and macrovascular complications. Perhaps most important among these is the increased risk of cardiovascular events in this patient population, an observation that can be best appreciated by the increased number of deaths attributed to cardiovascular-related diseases in diabetic patients aged 45 to 65 years. Consequently, aggressive therapy in this population offers the promise of significantly reducing excess cardiovascular deaths. Despite this opportunity for reducing mortality in these high-risk patients, several challenges to treatment remain. While aggressive blood pressure reduction has been documented to reduce the rate of events in these patients, questions remain as to the level to which blood pressure should be reduced. The recent guidelines from the Sixth Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure emphasized the importance of treating patients with hypertension and diabetes as if they already have target organ damage. Low blood pressure targets of 130/85 mm Hg, with an optimal goal of 120/80 mm Hg, can reduce the risk of events in hypertensive patients with diabetes, regardless of the pharmacological means used. However, there are physiologic and clinical rationale for renin angiotensin system blockade, with angiotensin-converting enzyme inhibition as the preferential therapy in these patients. In this regard, preliminary data with the new class of angiotensin II receptor blockers suggest that these agents may offer benefits equivalent to those observed with angiotensin-converting enzyme inhibitors while offering better tolerance. PMID- 10847252 TI - The attitudes of patients with advanced AIDS toward use of the medical futility rationale in decisions to forego mechanical ventilation. AB - BACKGROUND: The medical futility rationale asserts that physicians need not offer their patients therapies that have zero or close to zero probability of success. The rationale is controversial, but it is used in practice. OBJECTIVE: To examine the attitudes of patients with advanced acquired immunodeficiency syndrome (AIDS) toward the medical futility rationale as it might apply to their medical care. METHODS: We conducted a cross-sectional study with interviewer-administered questionnaires. Fifty-seven patients with advanced AIDS (C3 stage AIDS and a CD4 cell count <100/microL) were recruited from academic and private clinics. MAIN OUTCOME MEASURE: Whether patients believe it is acceptable for physicians to withhold mechanical ventilation, without offering it, if physicians determine the intervention is futile. RESULTS: Sixty-one percent of patients (n=35) believed that it was definitely acceptable for their physician to not offer mechanical ventilation if the physician judged this intervention to be futile, and 26% (n= 15) believed this was probably acceptable. Less than 10% of patients (n= 5) said not offering therapy judged futile was definitely not acceptable. Patients who were less likely to prefer mechanical ventilation in different hypothetical health states were significantly more likely to accept decisions on the basis of futility (P=.003). Health-related quality of life, patient satisfaction with medical care, and patient-clinician communication about end-of-life care were not associated with attitudes toward medical futility. CONCLUSIONS: Although the majority of patients with advanced AIDS accept the medical futility rationale, a substantial minority do not. Acceptance of this rationale was associated with wanting less life-sustaining treatment. Physicians invoking the medical futility rationale and hospitals using policies incorporating the medical futility rationale should take into account this diversity in the attitudes toward medical futility. PMID- 10847253 TI - Indications for hospitalization of patients with hyperkalemia. AB - BACKGROUND: Although the methods for the appropriate management of patients with hyperkalemia are well established, no criteria for hospital admission of patients with this common electrolyte disorder have been promulgated. OBJECTIVES: To examine the current practices regarding hospitalization of patients with hyperkalemia and to consider appropriate criteria for admission. PATIENTS AND METHODS: We evaluated a consecutive series of patients hospitalized for hyperkalemia and excluded patients who developed hyperkalemia after admission. For comparison, we selected a series of patients with a similar degree of hyperkalemia who were treated as outpatients. Hyperkalemia was classified as minimal, moderate, or severe. The causes of hyperkalemia were identified, and the therapeutic maneuvers used were ascertained. Although the study did not have the power to determine the relative safety of the 2 therapeutic approaches, we compared the outcomes of the 2 groups of patients. RESULTS: The inpatient group consisted of 11 patients who were admitted for the treatment of hyperkalemia, and we identified 12 patients who received outpatient therapy for hyperkalemia. The patients in the 2 treatment groups were similar with respect to age and the values of serum urea nitrogen, creatinine, and potassium prior to the identification of hyperkalemia. The mean +/-SD potassium concentrations at baseline were 5.4+/-0.7 mmol/L in the inpatients and 5.5+/-0.5 mmol/L in the outpatients. The mean +/-SD potassium concentration in the inpatients was 6.7+/ 0.8 mmol/L at the time of hospital admission, compared with 6.7+/-0.5 mmol/L in the outpatients at the time that hyperkalemia occurred. Similar proportions of both groups (6 of 11 inpatients and 7 of 12 outpatients) had moderate or severe hyperkalemia. CONCLUSIONS: Patients admitted to the hospital were clinically indistinguishable from patients treated as outpatients. The justification for the decision to admit patients to the hospital or to treat them as outpatients was often not evident. We suggest criteria for hospitalization, which include severe hyperkalemia (> or =8.0 mmol/L, with changes other than peaked T waves on the electrocardiogram), acute worsening of renal function, and supervening medical problems. PMID- 10847255 TI - All-cause mortality associated with physical activity during leisure time, work, sports, and cycling to work. AB - BACKGROUND: Physical activity is associated with low mortality in men, but little is known about the association in women, different age groups, and everyday activity. OBJECTIVE: To evaluate the relationship between levels of physical activity during work, leisure time, cycling to work, and sports participation and all-cause mortality. DESIGN: Prospective study to assess different types of physical activity associated with risk of mortality during follow-up after the subsequent examination. Mean follow-up from examination was 14.5 years. SETTING: Copenhagen University Hospital, Copenhagen, Denmark. PARTICIPANTS: Participants were 13,375 women and 17,265 men, 20 to 93 years of age, who were randomly selected. Physical activity was assessed by self-report, and health status, including blood pressure, total cholesterol level, triglyceride levels, body mass index, smoking, and educational level, was evaluated. MAIN OUTCOME MEASURE: All cause mortality. RESULTS: A total of 2,881 women and 5,668 men died. Compared with the sedentary, age- and sex-adjusted mortality rates in leisure time physical activity groups 2 to 4 were 0.68 (95% confidence interval, 0.64-0.71), 0.61 (95% confidence interval, 0.57-0.66), and 0.53 (95% confidence interval, 0.41-0.68), respectively, with no difference between sexes and age groups. Within the moderately and highly active persons, sports participants experienced only half the mortality of nonparticipants. Bicycling to work decreased risk of mortality in approximately 40% after multivariate adjustment, including leisure time physical activity. CONCLUSIONS: Leisure time physical activity was inversely associated with all-cause mortality in both men and women in all age groups. Benefit was found from moderate leisure time physical activity, with further benefit from sports activity and bicycling as transportation. PMID- 10847254 TI - Prospective study of the outcomes of ambulatory patients with excessive warfarin anticoagulation. AB - BACKGROUND: Warfarin sodium therapy is highly effective in preventing thromboembolism. Its major toxic effect is hemorrhage, the risk of which increases with the international normalized ratio (INR). Data on the rate of major hemorrhage and the rate of INR decay after an episode of excessive anticoagulation therapy would help guide management of elevated INRs in the outpatient setting. METHODS: We prospectively followed up outpatients in an anticoagulant therapy unit from April 24, 1995, through March 1, 1996. Study patients had to be taking warfarin for longer than 1 month and have an INR target range of 2.0 to 3.0. Consecutive outpatients with an INR greater than 6.0 were identified and compared with a randomly selected concurrent set of patients whose INR was in the target range. Major hemorrhage was defined as fatal, intracranial, or requiring hospitalization and transfusion of at least 2 U of blood. RESULTS: One hundred fourteen patients with INRs greater than 6.0 were identified and compared with 268 patients with INRs in the target range. None of the patients had clinically apparent bleeding at the time of the INR measurement, and none received phytonadione (vitamin K1). Patients did not differ significantly in age, sex, indication, or duration of warfarin therapy. Ten patients with an INR greater than 6.0 (8.8%; 95% confidence interval, 4.3%-15.5%) sought medical attention for abnormal bleeding, and 5 of these experienced a major hemorrhage during 14-day follow-up (4.4%; 95% confidence interval, 1.4%-9.9%) compared with none of the patients with an in-range INR (P<.001). Thirty-three percent of patients with INRs greater than 6.0 had INRs less than 4.0 within 24 hours, 55% within 48 hours, 73% within 72 hours, and nearly 90% within 96 hours of temporary discontinuation of warfarin therapy. CONCLUSIONS: Outpatients with INRs greater than 6.0 face a significant short-term risk of major hemorrhage. Randomized trials are needed to determine the net benefit of preventive treatment with phytonadione. PMID- 10847256 TI - The bleeding risk and natural history of idiopathic thrombocytopenic purpura in patients with persistent low platelet counts. AB - BACKGROUND: No firm data are available on the natural history of idiopathic thrombocytopenic purpura (ITP) or on mortality rates or frequency of major bleeding episodes associated with this condition. The disease is thought to have a relatively benign course, despite the frequent occurrence of very low platelet counts. This prevailing conception often guides therapeutic decisions. OBJECTIVE: To estimate the bleeding risk of ITP involving persistent low platelet counts (<30 x 10(9)/L) and its impact on prognosis. DESIGN: Age-adjusted bleeding risk was derived from a pooled analysis of ITP clinical series based on a systematic literature search. The risk estimate was incorporated into a Markov model to determine its impact on prognosis. RESULTS: Seventeen case series complied with inclusion criteria, including 1,817 patients with ITP. There were 49 cases of fatal hemorrhage over an estimated 1,258 to 3,023 patient-years at risk. The rate of fatal hemorrhage before age adjustment was estimated at between 0.0162 and 0.0389 cases per patient-year. Age-adjusted rates were 0.004, 0.012, and 0.130 cases per patient-year for age groups younger than 40, 40 to 60, and older than 60 years, respectively. Predicted 5-year mortality rates ranged from 2.2% for patients younger than 40 years to 47.8% for those older than 60 years. A 30-year old woman remaining thrombocytopenic due to ITP was predicted to lose 20.4 years (14.9 quality-adjusted life years) of her potential life expectancy. At age 70, predicted loss was 9.4 years (5.0 quality-adjusted life years). CONCLUSIONS: Idiopathic thrombocytopenic purpura with persistent low platelet counts carries a grave prognosis. Therefore, an active therapeutic approach in the clinical management of affected patients should be considered. In view of the significant potential implications of the model results, we call for initiating a well designed prospective inception cohort study of patients with ITP. PMID- 10847257 TI - Postmenopausal estrogen and estrogen-progestin use and 2-year rate of cognitive change in a cohort of older Japanese American women: The Kame Project. AB - BACKGROUND: The relation between estrogen and cognition among postmenopausal women remains controversial. Also uncertain is whether the proposed association varies between women taking unopposed estrogen and those taking estrogen combined with progestin. OBJECTIVE: To determine whether unopposed estrogen and combined estrogen-progestin use were associated with the rate of cognitive change in a cohort of older, Japanese American, postmenopausal women. METHODS: A prospective observational study in a population-based cohort of older Japanese Americans (aged > or =65 years) living in King County, Washington. Cognitive performance was measured in 837 women at baseline (1992-1994) and 2-year follow-up (1994 1997) examinations using the 100-point Cognitive Abilities Screening Instrument (CASI). Least squares means general linear models were used to estimate the 2 year rate of cognitive change according to categories of postmenopausal estrogen use. RESULTS: Approximately half of this cohort (n=455) had never used estrogen at any time since menopause, 186 were past users, 132 were current unopposed estrogen users, and 64 were current estrogen-progestin users. The majority of current estrogen users were taking conjugated estrogens, and all women receiving combined therapy were taking medroxyprogesterone acetate. After adjusting for age, education, language spoken at the interview, surgical menopause, and baseline CASI score, women who had never used postmenopausal estrogen improved slightly on the CASI scale (mean adjusted change, 0.79; SEM, 0.19). This change was significantly greater for current unopposed estrogen users (mean adjusted change, 1.68; SEM, 0.36; P=.04) and significantly worse for current estrogen progestin users (mean adjusted change, -0.41; SEM, 0.50; P =.02) compared with never users. The improvement observed in past users (mean adjusted change, 1.12; SEM, 0.29) was intermediate between the changes for never users and current unopposed estrogen users and not significantly greater than that for never users (P=.35). CONCLUSIONS: Our findings support a modest beneficial association between current unopposed estrogen use and the rate of cognitive change. We also observed a modest detrimental association between current estrogen-progestin use and the rate of cognitive change. The clinical significance of these modest differences, however, is uncertain. Data from large, long-term randomized trials are required before applying this information to the clinical setting. PMID- 10847258 TI - Headache in mild-to-moderate hypertension and its reduction by irbesartan therapy. AB - BACKGROUND: Although it is generally acknowledged to be a problem in severe hypertension, headache has not been consistently associated with mild-to-moderate hypertension. PATIENTS AND METHODS: In 7 randomized, double-blind, placebo controlled trials, which included 2,673 patients with mild-to-moderate hypertension (defined as seated diastolic blood pressure of 95-110 mm Hg), patients were randomized to receive once-daily treatment with irbesartan, an angiotensin II receptor blocker (n= 1,987), or placebo (n=686). The data were pooled and analyzed retrospectively to determine whether the level of hypertension was associated with headache and whether antihypertensive therapy reduced the incidence of headache. RESULTS: Factors found to be predictive of headache incidence were diastolic blood pressure, sex (female), and age (<50 years). In comparison with placebo, the use of irbesartan was associated with a significant reduction in the incidence of headache (P=.003). CONCLUSIONS: These data suggest that mild-to-moderate hypertension is not asymptomatic and that the incidence of headache can be reduced by antihypertensive treatment with a favorable adverse effect profile. PMID- 10847259 TI - Physical and sexual abuse in women infected with the human immunodeficiency virus: increased illness and health care utilization. AB - BACKGROUND: A history of physical or sexual abuse is associated with increased health care utilization and possibly an increase in illness based largely on self reported data. OBJECTIVE: To examine whether victimization experience is associated with increases in documented medical disease and health care utilization among women infected with the human immunodeficiency virus (HIV). METHODS: We interviewed and reviewed medical record data of 50 women seeking initial primary care for HIV infection at 2 urban hospitals. Women with and without a history of physical and sexual abuse were compared. Using multiple regression analysis, 2 periods were examined: entire life before study entry and the subsequent 2 years. The characteristics examined included episodic disease, chronic disease, sexually transmitted disease, chronic pain syndrome, opportunistic infections, obstetrical history, and number of injuries, surgical procedures, hospitalizations, ambulatory care visits, and emergency department visits. RESULTS: Evidence of physical or sexual abuse was found in 34 (68%) of the 50 women, of whom 16 (32%) did not disclose during the interview that they were abused. At entry, the rates of episodic disease, chronic pain syndrome, and sexually transmitted disease were greater among those women with histories of abuse. At 2 years, episodic disease, chronic disease, injuries, emergency department visits, and hospitalizations were all more likely in abused women. CONCLUSION: Physical and sexual abuse are common and associated with increased medical disease and health care utilization among HIV-infected women. PMID- 10847260 TI - Screening for hepatitis C virus in a health maintenance organization. AB - BACKGROUND: Chronic infection with hepatitis C virus (HCV) is a major public health problem and is associated with over 10,000 deaths a year in the United States. In its early stages, HCV tends to be asymptomatic and can be detected only through screening. OBJECTIVES: To develop and validate a database risk algorithm for HCV infection using electronic data at HealthPartners, a health maintenance organization (HMO) in Minnesota. A secondary objective was to evaluate the benefit of screening health care workers for HCV. METHODS: A database risk algorithm was developed using diagnostic and procedure codes in the administrative database to identify at-risk enrollees. One thousand three hundred eighty enrollees (an at-risk sample and a control sample) and 502 health care workers participated in anonymous screening. Both descriptive statistics and logistic regression were used to examine the frequency of HCV infection, associations with risk factors, self-selection factors in participation, and concordance between the database risk algorithm and the risk profile questionnaire. RESULTS: Eleven enrollees tested positive for HCV, 9 from the at risk sample and 2 from the control sample. All health care workers tested negative for HCV. Both lifestyle and medical risk factors were associated with positive test results for HCV. Enrollees with alcohol-drug diagnoses were less likely to participate in screening. A substantial proportion of enrollees with risk factors was identified either by the database risk algorithm or the risk profile questionnaire, but not by both. CONCLUSION: While the frequency of HCV infection was lower than previous estimates for the US population, the strong correlation with risk factors suggests that using the database risk algorithm for screening is a useful approach. Managed care plans with suitable data on their enrollee populations are in a key position to serve an important public health role in detecting asymptomatic patients who are infected with HCV. PMID- 10847261 TI - The efficacy of computer-tailored smoking cessation material as a supplement to nicotine polacrilex gum therapy. AB - BACKGROUND: Standard, generic self-help materials have been largely ineffective as behavioral treatments for smoking cessation. In contrast, self-help programs tailored to the needs of specific smokers have shown promise in facilitating quitting. OBJECTIVE: To evaluate the incremental efficacy of the Committed Quitters Program (CQP), a set of computer-tailored materials offered to purchasers of nicotine polacrilex gum, compared with a briefuntailored user's guide and audiotape, both as supplements to nicotine replacement therapy. METHODS: We conducted a randomized, open-label trial with 3 parallel arms. Subjects were smokers who purchased 2- or 4-mg nicotine polacrilex gum and called the CQP toll-free enrollment line. Three thousand six hundred twenty-seven subjects consented to participate in 1 of 3 study arms: (1) those receiving the CQP materials (CQP group, n= 1,217), (2) those receiving CQP materials and an outbound telephone call (CQP + C group, n= 1,207); and (3) those receiving no supplemental intervention beyond the user's guide and audiotape that were prepackaged with the nicotine polacrilex gum (UG group, n= 1,203). Twenty-eight day continuous abstinence rates were assessed by telephone interviews at 6 weeks and 10-week continuous rates at 12 weeks into treatment. RESULTS: Abstinence rates among respondents at the 6- and 12-week assessments were significantly higher for the CQP (36.2% and 27.6%) and CQP + C (35.5% and 27.3%) groups compared with the UG group (24.7% and 17.7%) at both intervals. The quit rates for the CQP and CQP + C groups were almost identical. CONCLUSIONS: The CQP proved to be an effective behavioral treatment, enhancing quit rates over and above nicotine replacement therapy and a brief untailored written guide and audiotape. PMID- 10847262 TI - Obstructive lung disease and low lung function in adults in the United States: data from the National Health and Nutrition Examination Survey, 1988-1994. AB - BACKGROUND: Obstructive lung disease (OLD) is an important cause of morbidity and mortality in the US adult population. Potentially treatable mild cases of OLD often go undetected. This analysis determines the national estimates of reported OLD and low lung function in the US adult population. METHODS: We examined data from the Third National Health and Nutrition Examination Survey (NHANES III), a multistage probability representative sample of the US population. A total of 20,050 US adults participated in NHANES III from 1988 to 1994. Our main outcome measures were low lung function (a condition determined to be present if the forced expiratory volume in 1 second-forced vital capacity ratio was less than 0.7 and the forced expiratory volume in 1 second was less than 80% of the predicted value), a physician diagnosis of OLD (chronic bronchitis, asthma, or emphysema), and respiratory symptoms. RESULTS: Overall a mean (SE) of 6.8% (0.3%) of the population had low lung function, and 8.5% (0.3%) of the population reported OLD. Obstructive lung disease (age-adjusted to study population) was currently reported among 12.5% (0.7%) of current smokers, 9.4% (0.6%) of former smokers, 3.1% (1.1%) of pipe or cigar smokers, and 5.8% (0.4%) of never smokers. Surprisingly, 63.3% (0.2%) of the subjects with documented low lung function had no prior or current reported diagnosis of any OLD. CONCLUSIONS: This study demonstrates that OLD is present in a substantive number of US adults. In addition, many US adults have low lung function but no reported OLD diagnosis, which may indicate the presence of undiagnosed lung disease. PMID- 10847263 TI - Why don't patients and physicians talk about end-of-life care? Barriers to communication for patients with acquired immunodeficiency syndrome and their primary care clinicians. AB - BACKGROUND: Patients with chronic and terminal disease frequently do not talk to their physicians about end-of-life care. Interventions to improve this communication have generally been unsuccessful, suggesting that important barriers to this communication must exist. OBJECTIVES: To determine the barriers to and facilitators of patient-clinician communication about end-of-life care and to identify barriers and facilitators that are more common among those patients who are least likely to discuss end-of-life care: minorities and injection drug users. METHODS: We conducted a prospective study of 57 patients with advanced acquired immunodeficiency syndrome and their primary care clinicians who were recruited from university and private clinics. Barriers to and facilitators of end-of-life communication were identified from a prior qualitative study and assessed for frequency and importance and for an association with the occurrence and quality of end-of-life communication. RESULTS: Clinicians identified more barriers than patients. Barriers identified by patients and clinicians fell into 3 categories of potential interventions: education about end-of-life care, counseling to help address end-of-life concerns, and health care system changes to facilitate patient-clinician communication. Although none of the patient identified barriers was associated with the occurrence of communication, 2 clinician-identified barriers were associated with less communication: "the patient has not been very sick yet" and "the patient isn't ready to talk about end-of-life care." Nonwhite patients were more likely to identify the following 2 barriers than white patients: "I feel that if I talk about death, it could bring death closer" and "I don't like to talk about the care I want if I get very sick." CONCLUSIONS: The diversity of barriers and facilitators relevant to patients with acquired immunodeficiency syndrome and their clinicians suggests that interventions to improve communication about end-of-life care must be focused on individual needs and must involve counseling interventions and health system changes in addition to education. Clinician barriers are more common and more strongly associated with the occurrence of end-of-life communication than patient barriers, suggesting that clinicians are an important target group for improving this communication. PMID- 10847264 TI - Evidence-based reduction of heart failure events with the involvement of pharmacists. PMID- 10847265 TI - Pneumococcal vaccination of the elderly: do we need another trial? PMID- 10847266 TI - Adverse drug reactions in the elderly: need for dedicated databases. PMID- 10847267 TI - We must save the art of medicine. PMID- 10847268 TI - Alendronate and nonsteroidal anti-inflammatory drug interaction safety is not established. PMID- 10847269 TI - Faculty and agora. PMID- 10847270 TI - A medical emancipation proclamation. PMID- 10847271 TI - Faculty should remain agoraphobic. PMID- 10847272 TI - HM-no! PMID- 10847273 TI - Managed care vs universal health insurance: whose whips are gentler? PMID- 10847274 TI - Valacyclovir hydrochloride therapy and thrombotic thrombocytopenic purpura in an HIV-infected patient. PMID- 10847275 TI - The will to live, suicide, and euthanasia. PMID- 10847276 TI - Magnetic resonance imaging in primary lymphoma of the spleen. PMID- 10847277 TI - Public perception of the dental degree. PMID- 10847278 TI - Distraction osteogenesis of the ascending ramus for mandibular hypoplasia using extraoral or intraoral devices: a report of 8 cases. AB - PURPOSE: This report presents the results of distraction osteogenesis using unidirectional extraoral and intraoral devices in 8 patients with different grades of vertical mandibular ramus hypoplasia. PATIENTS AND METHODS: Eight patients with hypoplastic mandibles underwent unilateral lengthening of the ascending ramus using unidirectional extraoral or intraoral devices. Intraoral mandibular distraction was performed on 5 patients with deficiencies of the vertical ramus up to 24 mm. External devices were used in 3 patients with more severe hypoplasias. An intraoral osteotomy was performed, and progressive distraction at rates of 0.5 mm/12 hours was initiated after 5 days. Once the desired length was reached, the device was maintained in place for 8 to 12 weeks. Three-dimensional computed tomography scans were taken in all the patients to plan the procedure and to compare the changes postoperatively. RESULTS: Successful distraction osteogenesis was achieved in all patients. The amount of mandibular lengthening ranged from 17 to 32 mm. Complications with the external devices such as rotation of the proximal bony fragment (2 cases) and loosening of the external screws at the end of the consolidation period (1 case) were observed. CONCLUSIONS: The results suggest that the intraoral device can be used as the method of choice for distraction osteogenesis of the ascending ramus of the mandible in patients with large deficiencies. Preoperative and postoperative 3-dimensional computed tomographic scans are essential in treatment planning. PMID- 10847279 TI - Management of the trochlea of the superior oblique muscle in the repair of orbital roof trauma. AB - PURPOSE: This study evaluated whether reattachment of the trochlea is necessary to assure normal postoperative extraocular muscle movements after orbital roof exploration and trochlea detachment. PATIENTS AND METHODS: This was a retrospective review of 889 operative records of one surgeon practicing at a level I trauma center between January 1, 1992 and December 31, 1998. Three hundred twenty-four of these cases were trauma related, 97 of which involved the upper facial third or upper midface. From this group, 15 patients required orbital roof exploration with trochlea detachment. Age, gender, cause of injury, form of craniofacial injury, method of repair, materials used, date of first notation of intact extraocular movement, and date of last follow-up were recorded and analyzed. RESULTS: From among the 15 patients requiring orbital roof exploration and repair, 14 (93%) were male, with a mean age of 34.7 +/- 15.2 years. Eight (53%) were involved in motor vehicle accidents, 3 (20%) in motorcycle accidents, and 4 (27%) in high-energy impacts. Most (95%) had injuries that included the frontal sinus, the naso-orbital-ethmoid region (60%), and the orbital rims (60%). After bitemporal flap reflection, careful subperiosteal dissection, supraorbital and supratrochlear nerve repositioning, and trochlea detachment, simple reapproximation of the orbital soft tissues to the reconstructed orbit resulted in satisfactory extraocular muscle movements in all cases. CONCLUSIONS: When careful subperiosteal dissection is used, simple reapproximation of the soft tissues adjacent to the reconstructed orbital roof, without reattachment of the trochlea, is all that was necessary to assure satisfactory extraocular muscle movements postoperatively. PMID- 10847280 TI - The team concept in mandibular reconstruction after ablative oncologic surgery. AB - PURPOSE: This study evaluated the efficiency and cost-effectiveness of using a reconstructive team for mandibular reconstruction. METHODS: An outcome-based retrospective review of 64 patients who had undergone microvascular fibular reconstruction of the mandible was performed. Operating room time, use of blood products, intraoperative fluid replacement, and hospital and intensive care unit stay were evaluated. Patients were divided into 2 groups on a chronologic basis. Group 1 was the first 34 consecutive patients and group 2 was the next 30 consecutive patients. RESULTS: Group 2 had a significantly shorter operating room time (P < .0001), hospital stay (P = .012), and used significantly less blood (P = .002) and colloid (P = .044) than group 1, resulting in significant cost savings, ($5,061.47/patient). Analysis showed no differences between groups demographically or for wound complications. CONCLUSIONS: Experience and the development of a team concept significantly decreased the cost for mandibular reconstruction with free fibula flaps without increasing wound complications. PMID- 10847281 TI - Investigation of infectious organisms causing pericoronitis of the mandibular third molar. AB - PURPOSE: The purpose of the study was to identify the most frequently encountered pyogenic organisms involved in pericoronitis to permit more targeted antibiotic therapy. PATIENTS AND METHODS: Pericoronal pockets of mandibular third molars from 37 patients showing symptoms of acute, severe pericoronitis were sampled and subjected to microbiologic analysis, including primary evaluation by phase contrast microscopy. To avoid overgrowth with faster-growing, less fastidious organisms, specimens were cultured on a wide variety of selective media (supporting growth of fastidious bacteria, protozoa, and fungi). RESULTS: Microscopic examination indicated spirochetes in 55% and fusiform bacteria in 84% of the samples. A total of 441 microorganisms were isolated and identified from the 37 cultured samples. Besides obligate anaerobic bacteria, including various Actinomyces and Prevotella species, a predominantly facultative anaerobic microflora was cultivated, that is, Streptococcus milleri group (78% of samples), Stomatococcus mucilaginosus (71%), and Rothia dentocariosa (57%). CONCLUSION: It was concluded that the Streptococci milleri group bacteria, well-known for their ability to cause suppurative infections, are most likely involved in the pathogenesis of acute severe pericoronitis of the lower third molar. PMID- 10847282 TI - Relationship between the extent of fracture and the degree of enophthalmos in isolated blowout fractures of the medial orbital wall. AB - PURPOSE: This study investigated the relationship between the extent of fracture and enophthalmos in blowout fractures of the medial orbital wall. PATIENTS AND METHODS: Nine patients with isolated blowout fractures of the medial orbital wall, confirmed by computed tomography scans, were evaluated. The area of fracture and the volume of herniated orbital tissue were determined from computed tomography scans using simple linear measurements. Each of the calculated values for area and volume were compared with the degree of enophthalmos to determine whether there was any significant relationship between them. RESULTS: Enophthalmos increased proportionally as the area of fracture or the volume of herniated orbital tissue increased (P < .05). The area of fracture and the volume of herniated orbital tissue associated with 2 mm of enophthalmos were 1.9 cm2 and 0.9 mL, respectively, as calculated from the regression curve. CONCLUSION: Enophthalmos of 2 mm or more, which is a frequent indication for surgery, can be expected when the area of fracture is 1.9 cm2 or more, or the volume of herniated orbital tissue is 0.9 mL or more. PMID- 10847283 TI - Single-dose vicoprofen compared with acetaminophen with codeine and placebo in patients with acute postoperative pain after third molar extractions. AB - PURPOSE: The purpose of this double-blind, randomized study was to compare the efficacy and safety of a single dose of the following medications: 2 tablets of Vicoprofen (ibuprofen 200 mg/hydrocodone 7.5 mg; Knoll Pharmaceutical Co, Mount Olive, NJ), 2 tablets ofp6 acetaminophen with codeine phosphate (acetaminophen 300 mg/codeine 30 mg), and 2 tablets of placebo in the management of moderate to severe postoperative dental pain after surgical extraction of at least one impacted mandibular third molar. PATIENTS AND METHODS: One hundred twenty-five patients (75 women, 50 men) participated in the study. The time of first perceptible pain relief and meaningful pain relief were measured using a stopwatch technique. Pain intensity and pain relief scores were recorded using standard verbal descriptors at 0.5, 1, 1.5, 2, 2.5, 3, 4, 5, 6, 7, and 8 hours after dosing. At the conclusion of the study, patients completed a global evaluation for the effectiveness of the study medication. RESULTS: Both active treatments were superior to placebo for all analgesic measures. Pain relief scores were significantly better for Vicoprofen than placebo throughout the study and significantly better than for acetaminophen with codeine from 2 through 8 hours after dosing. The duration of analgesia (time to remedication) was significantly longer for Vicoprofen (median, 5.50 hours) compared with acetaminophen with codeine (median, 3.03 hours) and placebo (median, 1.00 hours). Mean global evaluation for Vicoprofen was significantly better than for placebo and acetaminophen with codeine. Overall, there were no significant differences in the adverse event profile among the 3 treatment groups. CONCLUSIONS: Vicoprofen was found to be an effective postoperative analgesic medication in the management of acute postoperative dental pain. Its total analgesic effect, duration of analgesia, and global evaluation were superior to acetaminophen with codeine and placebo in this study model. PMID- 10847284 TI - Sensory impairment after resection of the mandible: a case report of 10 cases. AB - PURPOSE: In patients who have undergone mandibular resection for benign tumors involving the inferior alveolar nerve, some recovery of the neurosensory impairment in the lower lip and chin region may occur. This study investigated the extent of the residual neurosensory impairment to the lower lip and chin on the resected side. PATIENTS AND METHODS: Ten patients who underwent resection of ameloblastoma of the mandible involving the inferior alveolar nerve but not crossing the midline were evaluated by questionnaire and neurosensory testing for the extent of residual neurosensory impairment. RESULTS: Neurosensory tests showed that there was moderate recovery, especially in the sensation of light touch and temperature. Two-point discrimination and brush directional discrimination showed the poorest recovery. CONCLUSION: All patients suffered some degree of neurosensory deficit, but there was definitely some recovery, especially in patients younger than 16 years. Most of the patients had adapted well and had minimal physical and social problems as a result of the neurosensory deficit. PMID- 10847285 TI - Primary non-Hodgkin's lymphoma of the jaws: immunohistochemical and genetic review of 10 cases. AB - PURPOSE: Non-Hodgkin's lymphoma (NHL) comprises a group of malignant lymphoproliferative diseases characterized by clonal expansion of lymphocytes at various levels of ontogenetic development. The aim of this study was to review the immunohistochemical and cytogenetic features of 10 cases of NLH of the jaws to determine their respective derivation. PATIENTS AND METHODS: Histopathologic and immunohistochemical review of 10 cases of large-cell lymphomas of the jaws, together with Southern blot analysis of 2 of the cases, was performed and results compared with the findings in the literature. RESULTS: In the 10 cases studied, the average age of onset of the NHL was 51 years, with a male-to-female ratio of 3:2. Tumefaction was the first clinical sign of disease. Eight of 10 cases were high-grade, large-cell NHLs, centroblastic type. Two cases were high-grade, large cell NHL, immunoblastic type. CONCLUSION: The immunohistochemical and Southern blot data remain the principal laboratory aids in the diagnosis and characterization of NHL, and they provide critical information for guiding clinicians to the appropriate treatment protocol for these malignancies. PMID- 10847286 TI - Measurement of orbital volume by a 3-dimensional software program: an experimental study. AB - PURPOSE: This study reports a simple method with high accuracy for determining orbital volume from computed tomography (CT) scans. MATERIALS AND METHODS: The volume of 20 orbits was evaluated in 20 dry skulls by use of a 3-dimensional software program in General Electric High-Speed Advantage CT/I (Milwaukee, WI), and compared with the volume obtained by direct measurement using the water displacement method. Accuracy of volume measurement by this software program was assessed statistically by paired samples t-test. RESULTS: The mean volume was found to be 28.37 mL +/- 2.15 by direct impression and 28.41 mL +/- 2.09 by the software program. Volume difference between the 2 methods averaged 0.93 +/- 1.08 mL for each orbit (P < .01). The correlation between the techniques was found to be high (r = 0.887, P < .01). There was no significant volume discrepancy between the 2 methods. CONCLUSION: Measurement by the technique described is an easy and accurate method of assessing the volume of the orbit. PMID- 10847287 TI - An anatomic study of the lingual nerve in the third molar region. AB - PURPOSE: The aim of this study was to determine the location of the lingual nerve in the lower third molar region. PATIENTS AND METHODS: In this study, 669 nerves from 430 fresh cadavers were examined. Measurements on each cadaver were made using a micrometer caliper to determine the horizontal and vertical position of the lingual nerve in the lower third molar region. RESULTS: In 94 cases (14.05%), the nerve was above the lingual crest, and in 1 case (0.15%), the nerve was in the retromolar pad region. In the remaining 574 cases (85.80%), the mean horizontal and vertical distances of the nerve to the lingual plate and the lingual crest 2.06 +/- 1.10 mm (range, 0.00 to 3.20 mm) and 3.01 +/- 0.42 mm (range, 1.70 to 4.00 mm), respectively. In 149 cases (22.27%), the nerve was in direct contact with the lingual plate of the alveolar process. CONCLUSIONS: This study confirms the relatively unsafe position of the lingual nerve in relation to some oral and maxillofacial surgery procedures. PMID- 10847288 TI - Hair transplantation: a review and technique presentation. PMID- 10847289 TI - A survey of resident selection procedures in oral and maxillofacial surgery. AB - PURPOSE: This study was conducted to analyze the current procedures used in oral and maxillofacial surgery resident selection, to compare these selection procedures with those used 2 decades ago, to determine whether any differences exist in the selection procedures between 4-year certificate programs and programs that offer formal medical education, and to provide criteria to assist in the counseling of dental students on the application process for oral and maxillofacial surgery residencies. SUBJECTS AND METHODS: Questionnaires were sent to the 106 oral and maxillofacial surgery graduate training programs accredited by the American Dental Association. To provide for a more direct comparison to the study completed in 1976, the current questionnaire was developed, using the original survey as a model. It was divided into 5 sections: general information, information obtained from the formal application and letters of recommendation, the interview, the decision process, and a retrospective view of past decisions. The results were tabulated and the Pearson chi-square test was used to determine statistical significance when comparing the 4-year certificate programs to the programs that offer formal medical education. RESULTS: Seventy-one responses (75.5%) from nonmilitary programs were returned and analyzed. Thirty-nine responses represented dual-degree (MD) programs. Factors that were considered very important when judging a candidate's written application included dental school class rank (76.1%), dental school basic science grades (70.4%), and dental school clinical grades (63.4%). Dual-degree programs placed a greater emphasis on predental basic science grades (P < .01) and dental national board scores (P < .05). When asked about prior resident selection, 86.7% of the respondents said they would select 80% of their former residents again. In addition, 89.9% of the respondents were satisfied with their current selection process. CONCLUSIONS: The procedures used to select oral and maxillofacial surgery residents are relatively constant among programs. Although the dual-degree and 4-year certificate programs use the same criteria for resident selection, the dual-degree programs place greater emphasis on predental academic performance and on the results of the national dental boards. Criteria used 22 years ago to select residents are still applicable, but there has been a shift in the importance of some variables. PMID- 10847290 TI - Facial infection caused by Mycobacterium avium-intracellulare. PMID- 10847291 TI - Pseudoaneurysm after a routine transbuccal approach for bone screw placement. PMID- 10847292 TI - Infiltrating angiolipoma of the cheek: a case report and a review of the literature. PMID- 10847293 TI - Juvenile xanthogranuloma of the lip: case report and literature review. PMID- 10847294 TI - Synchronous occurrence of 2 histologically distinct parotid neoplasms. PMID- 10847295 TI - Partial facial nerve paralysis resulting from an infected mandibular third molar. PMID- 10847296 TI - Ultrastructural and biophysical studies of the bone tissue surrounding a hydroxyapatite-coated implant: case report. PMID- 10847297 TI - Epithelial-myoepithelial carcinoma of the parotid gland with multiple distant metastases: a case report. PMID- 10847298 TI - Statistical clarification. PMID- 10847299 TI - Dental spending increased. PMID- 10847300 TI - Suboptimal hemoglobin levels: do they impact patients and their therapy? Introduction. PMID- 10847301 TI - Hemoglobin level and anemia in radiation oncology: prognostic impact and therapeutic implications. PMID- 10847302 TI - Is anemia a problem for European cancer patients and treating oncologists? PMID- 10847303 TI - Optimal hemoglobin levels for cancer patients. PMID- 10847305 TI - The 7 habits of highly effective psychopharmacologists, part 2: begin with the end in mind. PMID- 10847304 TI - Suboptimal hemoglobin levels: do they impact patients and their therapy? Audience responses. PMID- 10847306 TI - Clozapine for treatment-refractory schizophrenia, schizoaffective disorder, and psychotic bipolar disorder: a 24-month naturalistic study. AB - BACKGROUND: The aim of this study was to evaluate the 24-month response to clozapine in patients with schizophrenia, schizoaffective disorder, or psychotic bipolar disorder. METHOD: Ninety-one psychotic patients with a principal DSM-III R diagnosis of schizophrenia (N = 31), schizoaffective disorder (N = 26), or bipolar disorder with psychotic features (N = 34) were treated naturalistically with clozapine at flexible dosages over a 24-month period. Improvement was assessed by the 18-item Brief Psychiatric Rating Scale and the Clinical Global Impressions-Severity of Illness scale. RESULTS: All patients showed significant improvement 24 months from intake (p < .001). Such an improvement was significantly greater among patients with schizoaffective disorder or bipolar disorder than in patients with schizophrenia (p < .05). The presence of suicidal ideation at intake predicted greater improvement at endpoint. CONCLUSION: Clozapine appears to be effective and relatively well tolerated in acute and long term treatment of patients with psychotic bipolar disorder or schizoaffective disorder who have not responded to conventional pharmacotherapies. PMID- 10847307 TI - The safety of olanzapine compared with other antipsychotic drugs: results of an observational prospective study in patients with schizophrenia (EFESO Study). Pharmacoepidemiologic Study of Olanzapine in Schizophrenia. AB - INTRODUCTION: Results of controlled clinical trials should be confirmed through safety and effectiveness studies in nonselected patient cohorts treated according to routine clinical practice. METHOD: Outpatients with schizophrenia (ICD-10 criteria) entered this prospective, naturalistic study when they received a new prescription for an antipsychotic drug. Treatment assignment was based on purely clinical criteria, as the study did not include any experimental intervention. Safety was evaluated through the collection of spontaneous adverse events and a specific questionnaire for extrapyramidal symptoms. Global clinical status was measured through the Clinical Global Impressions-Severity (CGI-S) and the Global Assessment of Functioning (GAF) scales. RESULTS: From the 2967 patients included, 2128 patients were treated with olanzapine as monotherapy or combined with other drugs (olanzapine group), and 821 were treated with other antipsychotic drugs as monotherapy or combined with other drugs (control group). There were no statistical differences between treatment groups at baseline regarding age, gender, disease duration, or severity of symptoms. Olanzapine was well tolerated and effective in this study. Overall incidence of adverse events was significantly lower in the olanzapine group compared with the control group (p < .001). Somnolence and weight gain were significantly more frequent in the olanzapine group, and akathisia, dystonia, extrapyramidal syndrome, hypertonia, hypokinesia, and tremor were significantly higher in the control group. Clinical improvement at endpoint, measured through the mean change in the CGI-S and the GAF, was significantly higher in the olanzapine group compared with the control group (p = .004). CONCLUSION: These results show that olanzapine is safe and effective in nonselected schizophrenic outpatients and are consistent with the efficacy and safety profile that olanzapine has shown in previous controlled clinical trials. PMID- 10847308 TI - Patients with schizophrenia at risk for excessive antipsychotic dosing. AB - BACKGROUND: Patient Outcomes Research Team treatment recommendations were used to investigate the relationship between patient characteristics and higher-than recommended dosages (> 1000 chlorpromazine equivalents [CPZe]) at discharge. METHOD: Inpatients who met the DSM-IV criteria for schizophrenia or schizoaffective disorder were recruited from 4 general hospitals. For those patients (N = 293) prescribed antipsychotics at discharge, chi-square tests and multiple regression analyses were used to assess the relationship between demographics, admission characteristics, comorbid diagnoses, and antipsychotic dosages. The relationship between clinical symptoms and antipsychotic dosage at discharge was also examined. RESULTS: Antipsychotic dosages conformed to treatment guidelines for approximately 65% of patients; 21% received doses in excess of recommended levels. African American patients and those with a history of psychiatric hospitalization were more likely to be prescribed discharge antipsychotic doses greater than 1000 CPZe. Hospital differences in antipsychotic management were also observed. Regression analyses indicated that higher-than recommended dosages found among African American patients could not be explained by differences in symptom levels at discharge. Patients with more thought disorder were also more likely to be prescribed antipsychotic dosages in excess of the recommended range. Compared with oral administration, depot administration increased the risk of excess dosage by a factor of 30. Controlling for method of administration reduced the impact of race to nonsignificance. CONCLUSION: These results replicate earlier findings that minority individuals are more likely to be prescribed dosages in excess of the recommended range and suggest that this pattern is due to higher use of depot injection in African American patients. Further research should examine how patient characteristics and institutional factors influence medication use. PMID- 10847309 TI - Personality changes in adult subjects with major depressive disorder or obsessive compulsive disorder treated with paroxetine. AB - BACKGROUND: Human and animal studies point to 3 dimensions of personality that change during pharmacotherapy with a selective serotonin reuptake inhibitor (SSRI). Specifically, harm avoidance has been found to decrease, social dominance has been found to increase, and hostility in social situations has been found to decrease with SSRI treatment. We sought to determine personality changes in subjects with either major depressive disorder (MDD) or obsessive-compulsive disorder (OCD) treated with paroxetine. We also sought to determine whether or not these personality changes were associated with disease state (MDD vs. OCD) or treatment response (responders vs. nonresponders). METHOD: Thirty-seven subjects diagnosed with either MDD or OCD (according to DSM-IV criteria) completed the Cattell 16 Personality Factor Inventory (16-PF) before and after treatment with paroxetine. Treatment response was defined as a Clinical Global Impressions Improvement rating of "much" or "very much" improved and a drop in Hamilton Rating Scale for Depression score of at least 50% for MDD or Yale-Brown Obsessive Compulsive Scale score of at least 30% for OCD. RESULTS: No significant differences were found between subjects with MDD and OCD in personality change with treatment. In the whole group, treatment responders had a greater decrease than nonresponders in 16-PF factors relating to harm avoidance. An increase in social dominance factors and a decrease in factors relating to hostility in social situations were found, but these changes were not significantly different between responders and nonresponders. CONCLUSION: These findings indicate that certain personality dimensions change with SSRI treatment and that some of these changes are independent of clinical treatment response. PMID- 10847310 TI - Mirtazapine substitution in SSRI-induced sexual dysfunction. AB - BACKGROUND: Sexual side effects are a common and bothersome reaction to selective serotonin reuptake inhibitors (SSRIs), frequently leading to cessation of treatment. Mirtazapine, an alpha2-adrenoceptor and serotonin-2/3 receptor antagonist, appears to cause few sexual problems. METHOD: Nineteen patients (12 women and 7 men), with SSRI-induced sexual dysfunction who were in remission from major depressive disorder (total Hamilton Rating Scale for Depression [HAM-D] score < or = 10), were switched to open-label mirtazapine for up to 6 weeks. Mirtazapine was titrated from 7.5 mg to 45 mg daily, as tolerated. Sexual functioning was measured weekly with the Arizona Sexual Experiences Scale (ASEX), and depression was measured weekly with the HAM-D. RESULTS: Eleven patients (58%) had a return of normal sexual functioning (mean +/- SD ASEX score = 12+/-3), and another 2 (11%) reported significant improvement in sexual functioning (mean ASEX score reduced from 24+/-1 to 20+/-0). All nineteen patients maintained their antidepressant response (HAM-D score after 6 weeks of mirtazapine = 6+/-3). The most commonly reported side effects (using moderate/severe rating on a symptom checklist) were initial sedation (N = 3), irritability (N = 6), and muscle soreness and stiffness (N = 3). Weight gain of 10 to 20 lb (4.5-9 kg) was seen in 3 patients (2 women and 1 man). CONCLUSION: Mirtazapine is an effective antidepressant for many patients experiencing SSRI-induced sexual dysfunction. PMID- 10847311 TI - Group therapy for patients with bipolar disorder and substance dependence: results of a pilot study. AB - BACKGROUND: The authors' goal was to pilot test a newly developed manual-based group psychotherapy, called Integrated Group Therapy (IGT), for patients with bipolar disorder and substance dependence. METHOD: In this open trial, patients with DSM-IV bipolar disorder and substance dependence (N = 45) were recruited in sequential blocks to receive either group therapy (N = 21) or 6 monthly assessments, but no experimental treatment (N = 24). RESULTS: When compared with patients who did not receive group therapy, patients who received IGT had significantly better outcomes on the Addiction Severity Index drug composite score (p < .03), percentage of months abstinent (p < .01), and likelihood of achieving 2 (p < .002) or 3 (p < .004) consecutive abstinent months. CONCLUSION: IGT is a promising treatment for patients with bipolar disorder and substance dependence, who have traditionally had poor outcomes. It is unclear, however, how much of the improvement among the group therapy patients is attributable to the specific content of the treatment. A study comparing this treatment with another active psychotherapy treatment is warranted. PMID- 10847312 TI - Treatment of binge-eating disorder with topiramate: a clinical case series. AB - BACKGROUND: Reduced appetite and weight loss were found in clinical trials of topiramate for epilepsy. Binge-eating disorder is characterized by recurrent episodes of binge eating that are not associated with regular use of inappropriate compensatory behaviors. Overweight and obesity may be common complications. To explore the effectiveness and tolerability of topiramate in binge-eating disorder, we describe the response of 13 consecutive outpatients with binge-eating disorder to naturalistic, open-label treatment with topiramate. METHOD: The response of 13 female outpatients with binge-eating disorder by DSM IV criteria to naturalistic, open-label treatment with topiramate (100-1400 mg/day) was reviewed. Response of binge-eating disorder symptoms was clinically assessed as none, mild, moderate, marked, or remission. Weight and side effects were also evaluated. RESULTS: All 13 patients had comorbid Axis I psychiatric disorders along with binge-eating disorder and were receiving psychotropic medications at the time of topiramate administration. After beginning topiramate treatment, 9 patients displayed a moderate or better response of binge-eating disorder symptoms that was maintained for periods ranging from 3 to 30 months (mean +/- SD = 18.7+/-8.0 months). Two other patients displayed moderate or marked responses that subsequently diminished. The remaining 2 patients had a mild or no response. The mean +/- SD weight of the 13 patients decreased from 99.3+/-26.4 kg to 87.5+/-20.4 kg (z = -2.4, df = 1, p = .02), but only 7 patients lost > or = 5 kg of weight. The mean topiramate treatment dose was 492.3+/-467.8 mg/day for all 13 patients. The mean topiramate dose was higher in patients who lost > or = 5 kg than in patients who lost < 5 kg. Also, topiramate dose correlated significantly with weight loss (p < .01). In general, topiramate was well tolerated, with neurologic side effects the most common. Of 3 patients who discontinued topiramate because of side effects, 2 resumed the drug at a later date without significant recurrence of these effects. CONCLUSION: Topiramate may be an effective treatment for binge-eating disorder. Controlled studies of topiramate in binge-eating disorder appear warranted. PMID- 10847313 TI - Melatonin improves sleep quality of patients with chronic schizophrenia. AB - BACKGROUND: Accumulating evidence indicates decreased melatonin levels in patients with schizophrenia. Insomnia, mainly difficulty in falling asleep at night, is commonly reported in this population. Association of insomnia with low or abnormal melatonin rhythms has been repeatedly documented. Melatonin is an endogenous sleep promoter in humans. We hypothesized that insomnia in patients with schizophrenia may be partially due to diminished melatonin output. In this study, we measured melatonin output in patients with chronic schizophrenia and assessed the effects of melatonin replacement on their sleep quality. METHOD: In a randomized, double-blind, cross-over, clinically based trial, 19 patients with DSM-IV schizophrenia who were treated with the normal treatment regimen were given melatonin (2 mg, controlled release) or placebo for 2 treatment periods of 3 weeks each with 1 week washout between treatment periods (7 weeks total). For measuring endogenous melatonin production, urine was collected from each patient every 3 hours between 9:00 p.m. and 9:00 a.m. Actigraphy was performed for 3 consecutive nights at the end of each period. Activity- and rest-derived sleep parameters were compared for the whole population with treatment arm as the intervening variable. A separate analysis was performed for patients subgrouped into high versus low sleep efficiency. RESULTS: All patients had low melatonin output. Melatonin replacement significantly improved rest-derived sleep efficiency compared with placebo (83.5% vs. 78.2%, p = .038) in this population. Improvement of sleep efficiency was significantly greater (p < .0014) in low efficiency (80% vs. 67%) than high-efficiency sleepers (88% vs. 90%). In addition, during melatonin therapy, tendencies toward shortened sleep latency (by 40 minutes, p < .056) and increased sleep duration (by 45 minutes, p < .078) were observed in low- but not high-efficiency sleepers. CONCLUSION: Melatonin improves sleep efficiency in patients with schizophrenia whose sleep quality is low. PMID- 10847314 TI - Modafinil augmentation of antidepressant treatment in depression. AB - BACKGROUND: Despite a relative lack of controlled data, stimulants are often used to augment antidepressant treatment in patients who have had only a partial response to first-line therapy. Modafinil is a novel psychostimulant that has shown efficacy in, and was recently marketed for, treating excessive daytime sleepiness associated with narcolepsy. The mechanism of action of modafinil is unknown, but, unlike other stimulants, the drug is highly selective for the central nervous system, has little effect on dopaminergic activity in the striatum, and appears to have a lower abuse potential. METHOD: In this retrospective case series, we describe 7 patients with DSM-IV depression (4 with major depression and 3 with bipolar depression) for whom we used modafinil to augment a partial or nonresponse to an antidepressant. The Hamilton Rating Scale for Depression was administered as part of routine clinical practice prior to treatment and at each subsequent visit. RESULTS: At doses of 100 to 200 mg/day, all 7 patients achieved full or partial remission, generally within 1 to 2 weeks. All patients had some residual tiredness or fatigue prior to starting modafinil, and this symptom was particularly responsive to augmentation. Side effects were minimal and did not lead to discontinuation of the drug in any of the patients. CONCLUSION: Modafinil appears to be a drug with promise as an augmenter of antidepressants, especially in patients with residual tiredness or fatigue. It is a particularly attractive alternative to other stimulants because of its low abuse potential and Schedule IV status. PMID- 10847315 TI - Medication continuation and compliance: a comparison of patients treated with clozapine and haloperidol. AB - BACKGROUND: This study compares medication continuation and regimen compliance with the atypical antipsychotic medication clozapine versus the conventional antipsychotic haloperidol. METHOD: Data from a 15-site double-blind, randomized clinical trial (N = 423) were used to compare patients with DSM-III-R schizophrenia assigned to clozapine or haloperidol in terms of duration of participation while taking the randomly assigned study drug (continuation) and the proportion of prescribed pills that were taken (compliance). Multiple regression analysis was used to determine the relationship of baseline characteristics and measures of clinical change to continuation for the entire sample and for patients assigned to each medication. RESULTS: Patients assigned to clozapine continued taking the study drug for a mean of 35.5 weeks as compared with only 27.2 among patients assigned to haloperidol (F = 4.45, df = 1,422; p = .0001). No differences were found between the groups in the proportion of prescribed pills that were returned at any timepoint. Among patients assigned to haloperidol, poorer continuation was associated with being older and greater continuation with receiving public support. Among patients on clozapine treatment, continuation was poorer among African American patients and greater among patients who showed reduced clinical symptoms and akathisia. Continuation with clozapine was greater even after adjusting for these factors. CONCLUSION: Continuation with medication is greater with clozapine than haloperidol and is partly explained by greater symptom improvement and reduced side effects. No differences were found in regimen compliance. PMID- 10847316 TI - Need for a new framework to understand the mechanism of all antipsychotics. PMID- 10847317 TI - Atypical antipsychotics for treatment of mixed depression and anxiety. PMID- 10847318 TI - Gender and bipolar illness. AB - BACKGROUND: For major depression and schizophrenia, gender differences have been reported in symptom expression and course of illness. Gender differences in bipolar disorder are becoming increasingly apparent, but have been less studied. Research data on these differences will help determine whether gender is important in influencing illness variables such as course, symptom expression, and likelihood of comorbidity. METHOD: Charts of 131 patients (63 women and 68 men) with a DSM-IV diagnosis of bipolar disorder admitted to the University of California Los Angeles Mood Disorders Program over a 3-year period were reviewed to gather data on demographic variables and course of illness and to assess differences in the illness across genders. RESULTS: No significant gender differences were found in the rate of bipolar I or bipolar II diagnoses, although women were overrepresented in the latter category. Also, no significant gender differences emerged in age at onset, number of depressive or manic episodes, and number of hospitalizations for depression. Women, however, had been hospitalized significantly more often than men for mania. Further, whereas bipolar men were significantly more likely than bipolar women to have a comorbid substance use disorder, women with bipolar disorder had 4 times the rate of alcohol use disorders and 7 times the rate of other substance use disorders than reported in women from community-derived samples. CONCLUSION: For bipolar disorder, course of illness variables such as age at onset and number of affective episodes of each polarity do not seem to differ across genders. Women, however, may be more likely than men to be hospitalized for manic episodes. While both men and women with the illness have high rates of comorbidity with alcohol and other substance use disorders, women with bipolar disorder are at a particularly high risk for comorbidity with these conditions. PMID- 10847319 TI - Estrogen replacement therapy: implications for postmenopausal women with end stage renal disease. AB - Little information is available about either the potential beneficial or harmful effects of estrogen replacement therapy in postmenopausal women with end-stage renal disease. Although evidence supports a role for estrogen replacement therapy in postmenopausal women in the prevention of cardiovascular disease and bone loss, possible improvement in cognitive function, and the relief of menopausal symptoms, these conclusions may not be applicable to patients with end-stage renal disease, since these studies have generally excluded such women. This issue is of considerable importance since cardiovascular causes account for more than 50% of the all-cause mortality in patients with end-stage renal disease. However, estrogen replacement therapy may also have untoward effects in patients with the disease, including an increased risk of dialysis access thrombosis and potentially worsening coronary artery disease in postmenopausal patients. Furthermore, dosing of estrogens needs to be done carefully since renal excretion is important for the elimination of estrogen metabolites. Low dose or alternate day dosing in addition to monitoring estrogen levels may be warranted when prescribing estrogen replacement therapy to women with end-stage renal disease. In this review, it is our objective to analyze the evidence published in the literature so far and to weigh the risks and benefits of estrogen therapy in postmenopausal women with end-stage renal disease. PMID- 10847320 TI - Towards targeted treatment of glomerulonephritis. PMID- 10847321 TI - Growth factor pathways in proliferative glomerulonephritis. AB - Growth factor production, glomerular cell proliferation and glomerular extracellular matrix expansion are prominent features of mesangioproliferative and crescentic glomerulonephritides. Recent studies have provided additional insights into the mechanisms of mesangial cell proliferation and interaction with extracellular matrix, and have focused on selective interruption of relevant mediators of injury. Studies into mitogenic signalling and cell cycle regulation in glomerular epithelial cells support a pathogenetic role for growth factors and glomerular epithelial cell proliferation in focal segmental glomerulosclerosis, and possibly membranous nephropathy. A better appreciation of growth factor pathways may lead to new treatment strategies for human glomerulopathies. PMID- 10847322 TI - Chemokine blockade as a therapy for renal disease. AB - Chemokines have been verified as inflammatory mediators of experimental nephritis by using anti-chemokine antibodies, chemokine receptor antagonists or targeted chemokine gene disruption. In human disease, chemokines are expressed in the kidney in association with inflammatory infiltrates. Chemokine blockade is thus a potentially novel approach to therapy of inflammatory renal disease. Several anti chemokine strategies are under development. Effective use of anti-chemokine therapy for renal disease will require a precise understanding of the unique roles that individual chemokines have in renal pathology, and the genetic factors that regulate chemokine expression. PMID- 10847323 TI - Regulatory peptides and their antagonists in nephropathies. AB - Many peptides influence renal function and structure in physiological and pathophysiological situations. Bioactive peptides that regulate renal function and structure encompass various substances including vasopeptides, growth factors, cytokines and peptide hormones. We highlight some novel concepts indicating that the vasoactive peptides angiotensin II and endothelin-1 play a major role in the progression of renal disease. These effects may be amplified by reduced concentration of counteracting natriuretic peptides. In addition, recent evidence suggests that peptides such as leptin, previously not considered to exert any renal effects, may be involved in renal pathophysiology under certain conditions. One of the most imperative tasks in nephrology is to develop innovative strategies to slow the progression of chronic renal disease. Interference with the renal action of bioactive peptides will certainly be part of this strategy. PMID- 10847324 TI - Pathology of progressive nephropathies. AB - The role of cell cycle regulatory proteins in progression is elucidated. Human renal biopsy data show amelioration and even regression of structural injury with interventions. Data implicate new mechanisms of the renin-aldosterone-angiotensin system in progressive injuries, including immune modulation and a direct effect of aldosterone on progression. New approaches and interventions that target these fibrotic responses show promising results. PMID- 10847325 TI - Advances in renal development. AB - Branching morphogenesis, mesenchymal cell condensation and mesenchymal-to epithelial conversion are key steps in kidney development and morphogenesis of several other organs. Review articles describing our current knowledge of the genetic and molecular regulation of these processes have been published. Therefore, this review focuses on the important question of cell lineage specification during kidney development. We describe how new insights are being made into the specification of kidney cells in the intermediate mesoderm, as well as specification of cells that will form the renal mesenchyme, ureteric duct, nephron tubule epithelium and renal vasculature. PMID- 10847326 TI - Prospective use of DNA microarrays for evaluating renal function and disease. AB - At the forefront of the revolution in human genomics is DNA microarray technology, which evaluates expression levels or genotypes of thousands of genes simultaneously, by means of miniaturization and parallel processing. Furthermore, advances in bioinformatics will result in the creation of large databases, which will require complex software programming for structural analysis. Over the next decade, DNA microarrays, combined with sophisticated informatics and genomic databases, will provide molecular fingerprints of disease processes and prognoses. This review provides an update on DNA microarray technology and its application to renal diseases. PMID- 10847327 TI - Molecular genetics of human hypertension. AB - The year 1999 saw considerable activity in the area of hypertension-related molecular genetics. Several new monogenic hypertensive disorders, as well as a monogenic form of hypotension, were elucidated. Molecular genetics has made significant inroads in explaining basic mechanisms of magnesium homeostasis. Linkage strategies have been applied in family studies, sib-pair analyses, and twin studies. More stringent criteria for association studies have been formulated. The 11 beta-hydroxysteroid dehydrogenase gene, the prostacyclin synthase gene, genes coding for variants in G proteins, and adrenergic receptor genes have received particular attention. On the horizon are better phenotyped patient and subject collectives, expanded genotyping with the availability of a 300,000 genome-wide single-nucleotide polymorphism map, multigenic studies in the form of metabolic control analyses, and new bioinformatic strategies including neural networks. PMID- 10847328 TI - Effects of lifestyle modification on treatment and prevention of hypertension. AB - Recently published observational epidemiologic studies and clinical trials have provided additional evidence that weight loss, dietary sodium reduction, moderation of alcohol consumption, physical activity, and potassium supplementation reduce blood pressure. Lifestyle modification should be recommended for the prevention and treatment of hypertension in the general population. PMID- 10847329 TI - Genes and renal disease. AB - The identification of genetic linkage between polymorphic markers and common kidney diseases, including focal and segmental glomerulosclerosis and diabetic nephropathy, clearly demonstrates that inherited factors contribute to renal failure susceptibility. These breakthroughs reveal the powerful contribution that molecular genetic techniques can make in the search for inherited factors that initiate renal failure and lead to its progression. Additionally, the environmental factors predisposing to nephropathy will be more readily detectable when evaluated in genetically similar populations. This manuscript reviews the developments in genetic epidemiology and molecular genetics of chronic renal failure. PMID- 10847330 TI - Hypertension in end-stage renal disease patients. AB - The prevalence of hypertension is extremely high in end-stage renal disease, and is a probable contributor to the epidemic of cardiovascular disease in end-stage renal disease. However, the paucity of prospective, randomized clinical trials makes it difficult to precisely define treatment strategies. Therefore, at present time the guidelines developed by the National Kidney Foundation's Cardiovascular Disease Task Force should be followed. PMID- 10847331 TI - Large multicentre hypertension trials. AB - The earlier large multicentre trials in hypertensive patients addressed questions of whether mild to moderate hypertension should be treated and whether similar approaches would be effective in elderly hypertensive patients or those with isolated systolic hypertension. The research focus of recent trials has now shifted to how rather than whether such patients should be treated. Trials such as the Hypertension Optimal Treatment study attempted to discern optimal targets for long-term blood pressure control. Although unsuccessful in this primary aim, they have established the safety of aggressive blood pressure lowering to diastolic targets of less than 80 mmHg as well as the safety and efficacy of a calcium entry blocker as a first line antihypertensive approach. The Captopril Prevention Project study and Swedish Trial in Old Patients with Hypertension-2 trial focussed on whether there might be specific antiatherosclerotic advantages of the newer agents (angiotensin converting enzyme inhibitors and calcium entry blockers) over conventional therapy in comparison studies with beta blockers and diuretics. Similar efficacy for cardiovascular outcomes appears to be emerging for each of the major classes of drugs with the degree of blood pressure lowering of prime importance in cardiovascular disease prevention. PMID- 10847332 TI - Bibliography. Current world literature. Renal immunology and pathology. PMID- 10847333 TI - Bibliography. Current world literature. Epidemiology and prevention. PMID- 10847334 TI - Molecular mechanisms of microtubular organelle assembly in Tetrahymena. AB - Thanks to recent technological advances, the ciliate Tetrahymena thermophila has emerged as an attractive model organism for studies on the assembly of microtubular organelles in a single cell. Tetrahymena assembles 17 types of distinct microtubules, which are localized in cilia, cell cortex, nuclei, and the endoplasm. These diverse microtubules have distinct morphologies, stabilities, and associations with specific Microtubule-Associated Proteins. For example, kinesin-111, a microtubular motor protein, is required for assembly of cilia and is preferentially targeted to microtubules of actively assembled, immature cilia. It is unlikely that the unique properties of individual microtubules are derived from the utilization of diverse tubulin genes, because Tetrahymena expresses only a single isotype of alpha- and two isotypes of 1-tubulin. However, Tetrahymena tubulins are modified secondarily by a host of posttranslational mechanisms. Each microtubule organelle type displays a unique set of secondary tubulin modifications. The results of systematic in vivo mutational analyses of modification sites indicate a divergence in significance among post-translational mechanisms affecting either alpha- or beta-tubulin. Both acetylation and polyglycylation of alpha-tubulin are not essential and their complete elimination does not change the cell's phenotype in an appreciable way. However, the multiple polyglycylation sites on 1-tubulin are essential for survival, and their partial elimination dramatically affects cell motility, growth and morphology. Thus, both high-precision targeting of molecular motors to individual organelles as well as organelle-specific tubulin modifications contribute to the creation of diverse microtubules in a single cytoplasm of Tetrahymena. PMID- 10847335 TI - The role of unconventional myosins in Dictyostelium endocytosis. AB - Dictyostelium discoideum is a simple eukaryote amenable to detailed molecular studies of the endocytic processes phagocytosis and macropinocytosis. Both the actin cytoskeleton and associated myosin motors are well-described and a range of mutants are now available that enable characterization of the role of the cytoskeleton in a range of cellular functions. Molecular genetic studies have uncovered roles for two different classes of Dictyostelium unconventional myosins in endocytosis. The class I myosins contribute to both macropinocytosis and phagocytosis by playing a general role in controlling actin-dependent manipulations of the actin-rich cortex. A class VII myosin has been shown to be important for phagocytosis. This brief review summarizes what is known about the role of these different myosins in both fluid and particle uptake in this system. PMID- 10847336 TI - Is Leishmania (Viannia) peruviana a distinct species? A MLEE/RAPD evolutionary genetics answer. AB - A set of 38 Leishmania stocks from the Andean valleys of Peru was characterized by both Multilocus Enzyme Electrophoresis (MLEE) and Random Amplified Polymorphic DNA (RAPD). Data were analyzed in terms of taxonomy and evolutionary genetics. Synapomorphic MLEE and RAPD characters, clear-cut clustering, and strong agreement between the phylogenies inferred from either MLEE or RAPD supported the view that Leishmania (Viannia) peruviana and Leishmania (Viannia) braziliensis correspond to two closely related, but distinct monophyletic lines (clades) and can therefore be considered as "discrete typing units" (DTUs). The question whether the L. (V.) peruxviana DTU deserves species status is dependent upon the desirability of it, in terms of epidemiological and medical relevance. A previous Orthogonal Field Alternating Gel Electrophoresis (OFAGE) analysis of the same L. (V.) peruviana isolates was published by Dujardin et al. (1995b). The data from the different markers (i.e. MLEE, RAPD and OFAGE) were compared by population genetics analysis. RAPD and OFAGE provided divergent results, since RAPD showed a strong linkage disequilibrium whereas OFAGE revealed no apparent departure from panmictic expectation. MLEE showed no linkage disequilibrium. Nevertheless, contrary to OFAGE, this is most probably explainable by the limited variability revealed by this marker in L. (V.) peruviana (statistical type II error). RAPD data were consistent with the hypothesis that the present L. (V.) peruviana sample displays a basically clonal population structure with limited or no genetic exchange. Disagreement between RAPD and OFAGE can be explained either by accumulation of chromosomal rearrangements due to amplification/deletion of repeated sequences, or by pseudo-recombinational events. PMID- 10847337 TI - Ultrastructural and biochemical characterization of promastigote and cystic forms of Leptomonas wallacei n. sp. isolated from the intestine of its natural host Oncopeltus fasciatus (Hemiptera: Lygaeidae). AB - Promastigote forms of a trypanosomatid were isolated from the third and fourth ventricles of the midgut and from the hindgut of the phytophagous hemipteran Oncopeltus fasciatus. Some individuals had adhered to its anterior region, close to the flagellar pocket, or to the flagellum up to four rounded aflagellated forms known as straphangers cysts. Scanning electron microscopy revealed that the flagellated forms presented a twisted cell body and a long flagellum, and the cysts, smaller than the parental promastigote, had a nascent flagellum. Transmission electron microscopy showed that promastigotes were typical, while cystic forms were ovoid dense cells devoid of a cyst wall, but presenting a cell coat, a special subpellicular region limited by a membrane unit, and a condensed cytoplasm. The kinetoplast-DNA fibrils appeared as dense spots and the condensed chromatin was arranged in a labyrinthic structure. Desmosome-like structures, observed in the region of adhesion of the precystic forms to the parental promastigote, could explain how cysts remain attached to the mother cell during the encystation process. Release of membranes from the surface of promastigotes and cysts seems to be correlated with the condensation of the cytoplasm during encystment. Morphological and isozyme analyses indicated that this trypanosomatid belongs to the genus Leptomonas. The molecular karyotype of this isolate was compared with that of a strain of Leptomonas oncopelti obtained from Oncopeltus varicolor by contour-clamped homogeneous electric field (CHEF) electrophoresis and revealed similar DNA banding patterns between 2,200-825 Kb, but not in lower bands (825-225 Kb). This suggested that the isolate from O. fasciatus and that from O. varicolor were not identical. Based on our findings we are describing Leptomonas wallacei n. sp. for our isolate from O. fasciatus. PMID- 10847338 TI - The effects of elevated temperatures and various time-temperature combinations on the development of Brachiola (Nosema) algerae N. Comb. in mammalian cell culture. AB - Nosema algerae Vavra and Undeen 1970, a microsporidian known to cause infection in mosquitoes, develops in mammalian cell cultures at 24-35 degrees C and in the tails and footpads of athymic mice. More recently it has been reported to grow at 38 degrees C in human cell culture. The present study is a two-part temperature/development examination. The first part examines the development of N. algerae in rabbit kidney cell culture at 29 degrees C, which permits the formation of functional spores within 72 h, and compares the effect of elevated temperatures (36.0, 36.5, 37 degrees C) on parasite development. At these elevated temperatures, N. algerae infects but undergoes only one or two proliferative divisions, with no evidence of sporogony by 72 h post-inoculation. During this time, however, the host cells continue to divide resulting in fewer infected cells over time and giving the appearance of a diminished parasitemia. Additionally, at 37 degrees C some organisms degenerate/hibernate by 72 h while others remain viable/active. It is not until 96 h that the parasites appear in large clusters of proliferative stages in the few host cells that are infected. By 120 h post-inoculation, proliferative cells, sporoblasts, and early spores are observed. These results suggest that elevated temperatures impede proliferation rates and the onset of sporogony. The second part of this study evaluates developmental changes in N. algerae when incubation temperatures and times are varied during parasite growth, resulting in abnormal parasite morphology. These abnormalities were not present when parasites were grown at constant temperature (29-37 degrees C). This report demonstrates that N. algerae can successfully develop at high temperatures (37 degrees C), justifying its taxonomic relocation to the genus Brachiola. PMID- 10847339 TI - High numbers of naked amoebae in the planktonic waters of a mangrove stand in southern Florida, USA. AB - This is the first study to examine the abundance of naked amoebae in the water column of a mangrove stand. A total of 37 different morphotypes was noted and at least 13 of these are probably new species. Over a one-year sampling interval, amoebae averaged 35,400 cells liter(-1) (range 2,000-104,000) by an indirect enrichment cultivation method. Densities in the upper end of this range are the highest ever reported for any planktonic habitat. Variation between samples was related to the quantity of suspended aggregates (flocs) in the water column emphasizing that amoebae are usually floc-associated. The study also showed that it is essential to disrupt floc material prior to withdrawing sample aliquots for the indirect counting method since several amoebae can occupy the interstices of aggregates. There is concern that indirect enumeration methods that require organisms to be cultured in the laboratory seriously underestimate the true count. A direct counting method using acridine orange staining and epifluorescence microscopy was attempted to assess the possible magnitude of the error associated with indirect counting. While this direct method had limitations, notably the difficulty of unambiguously differentiating between small amoebae and nanoflagellates, the results suggested that the indirect method gave estimates that were close to the true count (within a factor of two). Mangrove waters are rich in heterotrophic protozoa (approximately 3 x 10 liter( 1)) and while the heterotrophic flagellates are by far the dominant group, naked amoebae outnumber ciliates some 20-fold. The ecological consequences of high numbers of amoebae, particularly the common small forms less than 10 microm in length, need to be examined for these important coastal sites. PMID- 10847340 TI - Analysis of the conserved cysteine periodicity of Paramecium variable surface antigens. AB - The major surface antigens expressed by free-living and parasitic protozoa commonly contain repeating cysteine motifs. Despite the common occurrence of these repeats their functional significance remains largely unexplored. In this paper we investigate the conserved cysteine repeats within the variable surface antigens of Paramecium tetraurelia. We show that deletion of 2 entire repeating units or portions of repeats near the N-terminus does not prevent expression of the A51 variable surface antigen. Alteration of a single cysteine to serine residue also has no effect on A51 expression. In contrast, deletions near the C terminus of the protein have identified a small segment within the repeats that is required for expression on the surface. The required region contains a number of conserved amino acid residues, yet site-directed mutagenesis of two residues (serine and threonine to alanine) did not prevent expression. These studies demonstrate the feasibility of using deletion analysis to identify regions critical for the expression of cysteine-rich surface antigens. The relationship of these results to the structure and expression of cysteine-rich surface proteins in other protozoa is discussed. PMID- 10847341 TI - Phylogenetic identification of hypermastigotes, Pseudotrichonympha, Spirotrichonympha, Holomastigotoides, and parabasalian symbionts in the hindgut of termites. AB - The phylogenetic diversity of parabasalian flagellates was examined based on the sequences of small subunit ribosomal RNA genes amplified directly from the mixed population of flagellates in the hindgut of lower termites. In total, 33 representative sequences of parabasalids were recovered from eight termite species. Fluorescent-labeled oligonucleotide probes specific for certain sequences were designed and used for the in situ identification of parabasalian species by whole-cell hybridization. The hypermastigotes, Pseudotrichonympha grassii, Spirotrichonympha leidyi, and Holomastigotoides mirabile in the hindgut of Coptotermes formosanus, and Spirotrichonympha sp. and Trichonympha spp. in Hodotermopsis sjoestedti were identified. In the phylogenetic tree constructed, the sequences from the termites were dispersed within the groups of known members of parabasalids, reflecting the presence of diverse parabasalids in the hindgut of termites. There were three paraphyletic lineages of hypermastigotes represented by Pseudotrichonympha, Trichonympha, and Spirotrichonympha, in agreement with the morphology-based taxonomic groups. The analysis of the tree root suggested that the Pseudotrichonympha group is the most probable ancient lineage of parabasalids and that the Trichonympha group is the secondly deep branching lineage. The Spirotrichonympha group and the Trichomonadida may have emerged later. PMID- 10847342 TI - Characterization of Sarcocystis falcatula isolates from the Argentinian opossum, Didelphis albiventris. AB - Two isolates of Sarcocystis falcatula were obtained from the lungs of budgerigars (Melopsittacus undulatus) fed sporocysts from two naturally-infected South American opossums (Didelphis albiventris). The two isolates were designated SF-1A and SF-2A. Both isolates induced fatal infections in budgerigars. Both isolates underwent schizogony in African green monkey kidney cells. The structure of schizonts in the lungs of budgerigars was more variable than that observed in cell culture. The two isolates were identified as S. falcatula by the two species specific Hinf 1 restriction fragments dervied from digestion of a PCR amplification using primers JNB33/JNB54. Thus, the South American opossum, D. albiventris, is a definitive host for S. falcatula. PMID- 10847343 TI - Ultrastructural distribution of poly (A)+ RNA during Trypanosoma cruzi cardiomyocyte interaction in vitro: a quantitative analysis of the total mRNA content by in situ hybridization. AB - Ultrastructural in situ hybridization was used to visualize the spatial distribution of poly (A)+ RNA and quantitate its relative amount within different cellular compartments of cardiomyocytes after T. cruzi infection. The amount of poly (A)+ RNA remained about the same up to 24 h post-infection. In contrast, its content was reduced 65% after 72 h of interaction, showing a marked decrease in the cell cytoplasm. This decline in poly (A)+ RNA level in host cell cytoplasm was concomitant with intracellular proliferation of T. cruzi amastigotes. Thus, T. cruzi may affect host cell cytoplasmic mRNA stability, associated with the parasite's intracellular multiplication. PMID- 10847344 TI - Trypanosoma cruzi infection affects actin mRNA regulation in heart muscle cells. AB - We have previously described alterations in the cytoskeletal organization of heart muscle cells (HMC) infected with Trypanosoma cruzi in vitro. Our aim was to investigate whether these changes also affect the regulation of the actin mRNAs during HMC differentiation. Northern blot analysis revealed that alpha-cardiac actin mRNA levels increased during cell differentiation while beta-actin mRNA levels declined. Nonmuscle cells displayed beta-actin mRNA signal localized at the cell periphery, while alpha-cardiac actin mRNA had a perinuclear distribution in myocytes. Trypanosoma cruzi-infected cells showed 50% reduction in alpha cardiac actin mRNA expression after 72 h of infection. In contrast, beta-actin mRNA levels increased approximately 79% after 48 h of infection. In addition, in situ beta-actin mRNA was delocalized from the periphery into the perinuclear region. These observations support the hypothesis that Trypanosoma cruzi affects actin mRNA regulation and localization through its effect on the cytoskeleton of heart muscle cells. PMID- 10847345 TI - Phylogenetic relationships of Pleistophora-like microsporidia based on small subunit ribosomal DNA sequences and implications for the source of trachipleistophora hominis infections. AB - The microsporidian Trachipleistophora hominis was isolated in vitro from the skeletal muscle of an AIDS patient. Since its discovery several more cases of myositis due to Trachipleistophora have been diagnosed but the source of infection is unknown. Morphologically, T. hominis most closely resembles Pleistophora and Vavraia, which undergo polysporous sporogony in sporophorous vesicles, but differs from these genera in the mode of formation of sporoblasts and in the morphology of the sporophorous vesicles. Alignment and analyses of the small subunit ribosomal DNA sequences of T. hominis and several other polysporoblastic genera indicated that its closest phylogenetic relationships were with species of the genera Pleistophora and Vavraia, in line with morphological predictions. The type species of the latter two genera are Pleistophora typicalis and Vavraia culicis; these are parasites of fish and mosquitoes, respectively. These results suggest two possible routes and sources of infection to AIDS patients, these being perorally by ingestion of inadequately cooked fish or crustaceans or percutaneously during a bloodmeal taken by a haematophagous insect. Support for an insect source has been provided by recent detection of a microsporidium from mosquitoes in human corneal tissue. PMID- 10847346 TI - Phylogenetic analysis of the small subunit ribosomal RNA of Marteilia refringens validates the existence of phylum Paramyxea (Desportes and Perkins, 1990). AB - Marteilia refringens is recognized as one of the most significant pathogens of bivalve molluscs. The nucleotide sequence of the small subunit ribosomal RNA gene of Marteilia refringens is used to elucidate the phylogenetic position of the phylum Paramyxea. Genomic DNA was extracted from sporangia of Marteilia, purified from infected blue mussels, Mytilus edulis, and flat oysters, Ostrea edulis. The sequences obtained from Marteilia species purified from both oysters and mussels were identical. The sequence identity was confirmed by in situ hybridization using a DNA probe targeted to a variable region of the ribosomal DNA. The small subunit ribosomal RNA gene sequence of M. refringens is very different from all known sequences of eukaryotic organisms, including those of myxosporeans and haplosporeans. Therefore, the phylum Paramyxea should continue to be recognized as an independent eukaryotic phylum. PMID- 10847347 TI - Inter-laboratory comparison of the CD-1 neonatal mouse logistic dose-response model for Cryptosporidium parvum oocysts. AB - cryptosporidium parvum oocyst viability can be determined by vital dyes, in vitro excystation, and cell culture; however, neonatal mouse infectivity assays are the reference method. Unfortunately, there have been few efforts to standardize methods for infectivity assays thus casting a veil of uncertainty over the significance and comparability of results. In order to address this issue, two laboratories proficient in measuring oocyst infectivity conducted independent dose titration studies with neonatal CD-1 mice using standardized protocols and a well-characterized isolate of Cryptosporidium parvum. The resulting independent logistic dose-response models derived by regression analysis were compared with each other and with a published model. The comparisons showed these dose-response functions to be reproducible under standardized conditions. It is important to standardize mouse strain, age of mice at inoculation and necropsy, oocyst isolate, and age of oocysts. However, other factors, including methods used to detect infectivity and to count oocyst doses, appear less critical. Adopting a standardized assay for oocyst infectivity will provide both a basis for comparing data from various oocyst disinfection studies and a suitable platform for evaluating new or existing in vitro viability surrogates such as excystation, vital dyes or cell culture. PMID- 10847348 TI - Fluorescent in situ detection of Encephalitozoon hellem spores with a 6 carboxyfluorescein-labeled ribosomal RNA-targeted oligonucleotide probe. AB - A fluorescent in situ hybridization assay has been developed for the detection of the human-pathogenic microsporidian, Encephalitozoon hellem in water samples using epifluorescence microscopy. The assay employs a 19-nucleotide species specific 6-carboxyfluorescein-labeled oligonucleotide probe, HEL878F, designed to be complementary to the nucleic acid sequence 878-896, a highly variable segment of the 16S ribosomal RNA of E. hellem spores. The specificity of this probe for its ribosomal RNA target site was confirmed using RNA degradation, ribosomal RNA target site competition, and nucleotide base mismatch control probe assays. Furthermore, the specificity of the HEL878F oligonucleotide probe for E. hellem spores was established when it was evaluated on spores from all three species of the genus Encephalitozoon that had been seeded in reagent water and environmental water concentrates. The specificity of the HEL878F oligonucleotide probe was further corroborated when tested on algae, bacteria, and protozoa commonly found in environmental water. The study demonstrates the applicability of a fluorescent in situ hybridization assay using a species-specific fluorescent-labeled oligonucleotide probe for the detection of E. hellem spores in water samples. PMID- 10847349 TI - Descriptions and phylogenetic systematics of Myxobolus spp. from cyprinids in Algonquin Park, Ontario. AB - Eight species of Myxobolus were collected from four species of cyprinids in Algonquin Park, Ontario. On the basis of spore morphology, five of these species are described as new and two are redescribed. The evolutionary relationships among these eight species were studied using partial small subunit ribosomal DNA (ssu-rDNA) sequence data. The resulting cladograms, which were highly resolved and with strongly supported relationships, allowed for the evaluation of spore morphology, host specificity, and tissue tropism, criteria traditionally used in species identification. These criteria, recently criticized for creating artificial rather than natural taxonomic groupings, were evaluated for their reliability in the systematics of the species examined. The data showed that distantly related species often infect the same host and tissue, and that closely related species often occur in different hosts. Morphologically similar species are more closely related to each other and the taxonomy based on spore morphology is consistent with the relationships depicted in the phylogenies. These results suggest that spore morphology is better than host specificity and tissue tropism as a species character, as well as for determining evolutionary relationships among the species of Myxobolus examined. PMID- 10847350 TI - Sequence analysis of the Rhop-3 gene of Plasmodium yoelii. AB - The 110 kDa/Rhop-3 rhoptry protein of Plasmodium falciparum is non-covalently associated with two other proteins, the 140 kDa Rhop-1 and the 130 kDa Rhop-2. cDNAs encoding Rhop-3 from Plasmodium yoelii were isolated using rhoptry-specific antisera from Plasmodium falciparum, P. yoelii, and Plasmodium chabaudi. The cDNAs encoded peptides with partial homology to the C-terminal region (residues 541-861) of P. falciparum Rhop-3. Core regions of homology to the P. falciparum gene will be useful in determining the biological role of Rhop-3 and its potential as a vaccine candidate for malaria. PMID- 10847351 TI - New facility picture archiving and communication system implementation strategy. AB - Strategies for deployment of picture archiving and communications systems (PACS) in new hospitals often involve the establishment of initial PACS operations. Such a strategy is flawed in the sense that the clinical and radiological users must adapt to PACS operations, while being faced with several other new facility learning curves. This increases the complexity and risk of the radiological services. A strategy of implementing PACS operations in the old facility and performing a zero-downtime transition into the new facility offers several advantages to this method. The successful undertaking of such a project will support not only the physical movement of the existing PACS, but the accomplishment of other re-engineering goals associated with the new hospital. This report will describe the strategy used in two successful transitions of PACS into newly constructed hospitals. PMID- 10847352 TI - How to effectively use consultants. AB - Apart from a few pioneering medical centers, most medical organizations lack the internal knowledge and technical expertise to develop a comprehensive digital imaging business plan, create medical imaging architectures, select vendor(s), and implement picture archiving and communication systems (PACS) and other information systems. Many of these processes are only done once (eg, vendor selection and initial implementation) and developing such one-time expertise in house is wasteful and inefficient. It is highly unlikely that an inexperienced hospital-based group undertaking their first and only PACS project will be able to match the experience and depth of a well-organized consulting group with prior experience. Organizations therefore are looking to consulting groups to fill their internal resource gaps. This report presents the benefits of engaging consultants, guidelines for selecting a consulting group, and insight on effectively using the consultants. The presentation is based on actual experiences using consultants for planning, selecting, and implementing a PACS at Children's Hospital Medical Center, Cincinnati. PMID- 10847353 TI - How to satisfy both clinical and information technology goals in designing a successful picture archiving and communication system. AB - Designing and operating a PACS system requires an integrated focus to maintain peak performance of the system from an information technology (IT) perspective and to ensure that all clinical and financial requirements are met. An IT-based picture archiving and communication system (PACS) manager is in the best position to satisfy these sometime conflicting audiences. This report will describe how an institution moving towards PACS can unite radiologists, hospital administrators, and information systems (IS)/IT specialists into one cohesive team to ensure the highest levels of success with their future PACS. There are several keys to success: (1) Designing and selecting PACS requires a dedicated team, with representatives from radiology, as well as IS/IT and administration. (2) Each group needs to thoroughly outline their specific needs, so that the final PACS solution is relevant from all perspectives. This needs assessment needs to be made before issuing a request for proposal (RFP) and interviewing vendors. (3) The team needs to be small to be effective. Each group should have one or at most two representatives that collect input from, and report to, a group of his or her peers. (4) Plans need to be made to determine how to integrate current and future hospital information systems (HIS), in order to ensure a smooth pathway to the electronic medical record. (5) All team members should agree on the overall objectives for PACS and participate in its design and installation. (6) Each team member is charged with motivating, and helping to educate, his or her peers. (7) Training should be tailored to the needs of each audience. Explain how each staff member benefits from the PACS. Training should be ongoing to accommodate the addition of new system features and new users. This report will describe the importance of recognizing PACS as being an IT system with a clinical focus. The importance of designing goals of the PACS system from various perspectives, including clinical, technical, and financial, will be addressed. More importantly, this presentation will highlight the benefits a medical institution will receive if the various groups can work together, while at the same time outlining some pitfalls they can expect to encounter if the groups take an adversarial approach. PMID- 10847354 TI - User acceptability--a critical success factor for picture archiving and communication system implementation. AB - The Department of Diagnostic Imaging at the Hospital for Sick Children (HSC), Toronto, implemented a picture archiving and communication system (PACS) during the last year. This report describes our experience from the point of view of user acceptability. Based on objective data, the following key success factors were identified: user involvement in PACS planning, training, technical support, and rollout of pilot projects. Although technical factors are critical and must be addressed, the main conclusion of our study is that other nontechnical factors need to be recognized and resolved. Recognition of the importance of these factors to user acceptance and clear communication and consultation will help reduce negative user attitudes and increase the chance of a successful PACS implementation. PMID- 10847356 TI - The filmless radiology reading room: a survey of established picture archiving and communication system sites. AB - The purpose of this study was to survey radiologists experienced in soft-copy diagnosis using computer workstations about their current reading room environment, their impressions of the efficacy of their reading room design, and their recommendations based on their experience for improvement of the soft-copy reading environment. Surveys were obtained from radiologists at seven sites representing three major picture archiving and communication system (PACS) vendors throughout the world that have had extensive experience with soft-copy interpretation of radiology studies. The radiologists filled out a detailed survey, which was designed to assess their current reading room environment and to provide them with the opportunity to make suggestions about improvement of the PACS reading rooms. The survey data were entered into a database and results were correlated with multiple parameters, including experience with PACS, types of modalities interpreted on the system, and number of years of experience in radiology. The factors judged to be most important in promoting radiologist productivity were room lighting, monitor number, and monitor brightness. Almost all of the radiologists indicated that their lighting source was from overhead rather than indirect or portable light sources. Approximately half indicated they had the capability of dimming the brightness of the overhead lighting. Most radiologists indicated that they were able to adjust room temperature but that they did not have individual temperature controls at their workstations. The radiologists indicated that the most troublesome sources of noise included background noise, other radiologists, and clinicians much more than noise from computer monitors, technologists, or patients. Most radiologists did not have chairs that could recline or arm rests. Most did have wheels and the capability to swivel, both of which were judged important. The majority of chairs also had lumbar support, which was also seen to be important. Radiologists commonly adjusted room lighting and their reading chair, but rarely adjusted room temperature or monitor brightness. The median number of hours spent at the workstation before taken a "break" was 1.5. Common recommendations to improve the room layout included compartmentalization of the reading room and availability of the hospital/radiology information system at each workstation. The survey data suggest several areas of potential improvement based on radiologists' experience. Optimization of soft-copy reading room design is likely to result in decreased fatigue and increased productivity. PMID- 10847355 TI - New aspects of image distribution and workflow in radiology. AB - The progressive use of digital image-generating devices and digital communication technology in clinical and practice environments implies changes in radiological workflow and asks for adequate quality assurance in the whole process of radiology report preparation. This improvement potential has to be rigorously reinvestigated with regard to up-to-date procedures and the full exploitation of supporting technologies like linguistic analysis, help desk and trouble ticket systems, competitive allocation algorithms, time-and-event monitoring, and intelligent agents. These approaches are to be evaluated in combination with business process analysis and shall help to reduce turnaround times for radiology reports while maintaining or even increasing quality-assurance levels. PMID- 10847357 TI - Use of three-dimensional spiral computed tomography imaging for staging and surgical planning of head and neck cancer. AB - We compare four different three-dimensional (3D) reconstruction methods of spiral computed tomography (CT) data for head and neck cancer to establish the method best suited for specific uses, eg, staging of lymph nodes and viewing of spatial relationships between the tumor, fascial spaces, adjacent soft tissues, and others structures. We evaluated a series of 10 patients (six men and four women), aged 32 to 60 years. Of these, five were histologically diagnosed with squamous cell carcinoma, two with lymphoma, one with thyroid cancer, one with Kikuchi's disease or necrotizing lymphadenitis, and one with esthesioneuroblastoma. All scans were obtained using high-resolution spiral CT (General Electric Medical Systems, Milwaukee, WI). The collimations used were 3 mm and 5 mm, matrix 512 x 512, and reconstruction interval not more than 3 mm. Scanning was performed from the skull base to the aortic arch. Iodinated contrast medium was injected so that the blood vessels were clearly differentiated from nodes. Different techniques of three-dimensional reconstruction were employed, including shaded surface display (SSD), multiplanar reconstructions (MPR), maximum intensity projection (MIP), 3D volume rendering (VR), and combined techniques. The reconstructions were performed in a variety of planes, including sagittal, coronal, and oblique views. In our series of selected patients, the technique of 3D VR showed potential advantages over other techniques. The MIP technique was useful in analyzing the patency of vessels and to exclude thrombus, compression, or displacement by tumor. The use of combined techniques such as SSD and MPR, accurately demonstrated the levels of lymph nodes and the relationship between the tumor projection of interest and various anatomic structures. In conclusion, 3D reconstruction of CT data is useful in the localization and staging of neck tumors and assists in surgical planning and radiation treatment. PMID- 10847358 TI - Image compression and chest radiograph interpretation: image perception comparison between uncompressed chest radiographs and chest radiographs stored using 10:1 JPEG compression. AB - We have assessed the effect of 10:1 lossy (JPEG) compression on six board certified radiologists' ability to detect three commonly seen abnormalities on chest radiographs. The study radiographs included 150 chest radiographs with one of four diagnoses: normal (n = 101), pulmonary nodule (n = 19), interstitial lung disease (n = 19), and pneumothorax (n = 11). Before compression, these images were printed on laser film and interpreted in a blinded fashion by six radiologists. Following an 8-week interval, the images were reinterpreted on an image display workstation after undergoing 10:1 lossy compression. The results for the compressed images were compared with those of the uncompressed images using receiver operating characteristic (ROC) analyses. For five of six readers, the diagnostic accuracy was higher for the uncompressed images than for the compressed images, but the difference was not significant (P > .1111). Combined readings for the uncompressed images were also more accurate when compared with the compressed images, but this difference was also not significant (P = .1430). The sensitivity, specificity, and accuracy values were 81.5%, 89.2%, and 86.7% for the compressed images, respectively, as compared with 78.9%, 94.5%, and 89.3% for the uncompressed images. There was no correlation between the readers' accuracy and their experience with soft-copy interpretation; the extent of radiographic interpretation experience had no correlation with overall interpretation accuracy. In conclusion, five of six radiologists had a higher diagnostic accuracy when interpreting uncompressed chest radiographs versus the same images modified by 10:1 lossy compression, but this difference was not statistically significant. PMID- 10847359 TI - Shape analysis of hippocampal surface structure in patients with unilateral mesial temporal sclerosis. AB - Structural hippocampal magnetic resonance (MR) imaging-based analysis is helpful in the diagnosis and treatment of mesial temporal epileptic seizures. Computational anatomic techniques provide a framework for objective assessment of three-dimensional hippocampal structure. We applied a previously validated technique of deformation-based hippocampal segmentations in 20 subjects with documented unilateral mesial temporal sclerosis (MTS) and temporal lobe epilepsy. Using composite images, we then measured shape differences between the epileptogenic, smaller hippocampus, and contralateral hippocampus. Final shape differences were projected on the contralateral "normal" side. We calculated results for the left MTS group (10 patients) and right MTS group (10 patients) separately. Both groups showed similar regions of maximal inward deformation in the affected hippocampus, which were the medial and lateral aspect of the head, and posterior aspect of the tail. These results suggest that there are specific three-dimensional patterns of volume loss in patients with mesial temporal epilepsy. PMID- 10847360 TI - Computed radiography printing problems: a quantitative, observer-independent solution. AB - Even though facilities using computed radiography (CR) operate in an electronic environment, the production of hard-copy films is still necessary during the transition period, as well as for particular needs following complete implementation. We have implemented a quantitative technique to match the response of printed CR film with that of previous screen/film combinations. A stepwedge is radiographed using the conventional system. The same stepwedge is then radiographed (same geometry and technique) using the CR system. Following processing and printing, the plot of optical density versus step for the CR system is compared with that of the screen/film system. Adjustments are made to the printing parameters until the response curves are identical. All other translation tables in the system are set to be linear. This has proven to be a valuable technique for us and provides CR printed image quality that is equivalent to that of our previous screen/film combinations. PMID- 10847361 TI - Radiologist's clinical information review workstation interfaced with digital dictation system. AB - Efficient access to information systems integrated into the radiologist's interpretation workflow will result in a more informed radiologist, with an enhanced capability to render an accurate interpretation. We describe our implementation of radStation, a radiologist's clinical information review workstation that combines a digital dictation station with a clinical information display. radStation uses client software distributed to the radiologist's workstation and central server software, both running Windows NT (Microsoft, Redmond, WA). The client system has integrated digital dictation software. The bar-code microphone (Boomerang, Dictaphone Corp, Stratford, CT) also serves as a computer input device forwarding the procedure's accession number to the server software. This initiates multiple queries to available legacy databases, including the radiology information system (RIS), laboratory information system, clinic notes, hospital discharge, and operative report system. The three-tier architecture then returns the clinical results to the radStation client for display. At the conclusion of the dictation, the digital voice file is transferred to the dictation server and the client notifies the RIS to update the examination status. The system is efficient in its information retrieval, with queries displayed in about 1 second. The radStation client requires less than 5 minutes of radiologist training in its operation, given that its control interface integrates with the well-learned dictation process. The telephone-based dictation system, which this new system replaced, remains available as a back-up system in the event of an unexpected digital dictation system failure. This system is well accepted and valued by the radiologists. The system interface is quickly mastered. The system does not interrupt dictation workflow with the display of all information initiated with examination bar-coding. This system's features could become an accepted model as a standard tool for radiologists. PMID- 10847362 TI - Enhancing the expressiveness of structured reporting systems. AB - The overall goal of this research is to build a structured reporting system that reduces the cost, delays, and inconvenience associated with conventional dictation and speech recognition systems. We have implemented such a structured reporting system for radiology that replaces current dictation and transcription processes by allowing radiologists and other imaging professionals to select imaging findings from a medical lexicon. The system uses an imaging-specific information model, called a "description set,' to organize selected terms in a relational database. Unique features of the knowledge representation that enhance its expressiveness include its ability to codify uncertainty about an imaging observation and to represent explicitly the logical relationships among imaging findings. In addition, the system does not require the user to fill in "blanks' in a static text template. Instead, it allows entry of terms in arbitrary order and uses automated text-generation techniques to create a text report that referring physicians are accustomed to receiving. In parallel, the system also produces a multimedia report that the referring physician can use as a quick reference. Unlike the results of conventional dictation or speech recognition, each finding is coded in a relational database for later information processing. Thus, the structured report database can be used to index images by content, to provide real-time decision support, to enhance radiologists' performance, to conduct exploratory clinical research, and to transmit imaging report data to computer-based patient record systems. PMID- 10847363 TI - Continuing professional development for radiologists using the Web as a delivery vehicle. PMID- 10847364 TI - American Board of Radiology computer test center. AB - In 1997, the American Board of Radiology (ABR) determined to develop a computer based examination and to create a test center for administration of computer based examinations. In implementation of its plan, the Board has developed a flexible examination platform, well-adapted to the graphics needs of an image based examination, and at the same time, compatible with test centers being developed by other medical specialty boards in terms of hardware, software, and candidate surroundings. A test center for secure proctored examination of up to 33 candidates has been created at the Board's headquarters in Tucson, AZ. The decision of the ABR to employ computer-based testing as a part of its recertification process represents an important step of significance to the entire field of radiology, embracing methods that are rapidly becoming integral to the practice of radiology in the acquisition, display, and management of diagnostic imaging information. PMID- 10847365 TI - Common object request broker architecture (CORBA)-based security services for the virtual radiology environment. AB - The US Army Great Plains Regional Medical Command (GPRMC) has a requirement to conform to Department of Defense (DoD) and Army security policies for the Virtual Radiology Environment (VRE) Project. Within the DoD, security policy is defined as the set of laws, rules, and practices that regulate how an organization manages, protects, and distributes sensitive information. Security policy in the DoD is described by the Trusted Computer System Evaluation Criteria (TCSEC), Army Regulation (AR) 380-19, Defense Information Infrastructure Common Operating Environment (DII COE), Military Health Services System Automated Information Systems Security Policy Manual, and National Computer Security Center-TG-005, "Trusted Network Interpretation." These documents were used to develop a security policy that defines information protection requirements that are made with respect to those laws, rules, and practices that are required to protect the information stored and processed in the VRE Project. The goal of the security policy is to provide for a C2-level of information protection while also satisfying the functional needs of the GPRMC's user community. This report summarizes the security policy for the VRE and defines the CORBA security services that satisfy the policy. In the VRE, the information to be protected is embedded into three major information components: (1) Patient information consists of Digital Imaging and Communications in Medicine (DICOM)-formatted fields. The patient information resides in the digital imaging network picture archiving and communication system (DIN-PACS) networks in the database archive systems and includes (a) patient demographics; (b) patient images from x-ray, computed tomography (CT), magnetic resonance imaging (MRI), and ultrasound (US); and (c) prior patient images and related patient history. (2) Meta-Manager information to be protected consists of several data objects. This information is distributed to the Meta-Manager nodes and includes (a) radiologist schedules; (b) modality worklists; (c) routed case information; (d) DIN-PACS and Composite Health Care system (CHCS) messages, and Meta-Manager administrative and security information; and (e) patient case information. (3) Access control and communications security is required in the VRE to control who uses the VRE and Meta-Manager facilities and to secure the messages between VRE components. The CORBA Security Service Specification version 1.5 is designed to allow up to TCSEC's B2-level security for distributed objects. The CORBA Security Service Specification defines the functionality of several security features: identification and authentication, authorization and access control, security auditing, communication security, nonrepudiation, and security administration. This report describes the enhanced security features for the VRE and their implementation using commercial CORBA Security Service software products. PMID- 10847366 TI - A centralized dose calculation system for radiation therapy. AB - Centralization of treatment planning in a radiation therapy department is a realistic strategy to achieve an integrated and quality-controlled planning system, especially for institutions with numerous affiliations. The rapid evolution of computer hardware and software technology makes this a distinct possibility. However, the procedure of three-dimensional treatment planning involves a number of steps, such as: (1) input of patient computed tomography (CT) images and contour information; (2) interactions with local devices such as a film digitizer; and (3) output of beam information to be integrated with the record and verify the system. A full-fledged realization of the web-based centralized three-dimensional treatment planning system will require an extensive commercial development effort. We have developed and incorporated a web-based Timer/Monitor Unit (MU) program as a first step towards the full implementation of a centralized treatment planning system. The software application was developed in JAVA language. It uses the internet server and client technology. With one server that can handle multiple threads, it is a simple process to access the application anywhere on the network with an internet browser. Both the essential data needed for the calculation and the results are stored on the server, which centralizes the maintenance of the software and the storage of patient information. PMID- 10847367 TI - Relevant priors prefetching algorithm performance for a picture archiving and communication system. AB - Proper prefetching of relevant prior examinations from a picture archiving and communication system (PACS) archive, when a patient is scheduled for a new imaging study, and sending the historic images to the display station where the new examination is expected to be routed and subsequently read out, can greatly facilitate interpretation and review, as well as enhance radiology departmental workflow and PACS performance. In practice, it has proven extremely difficult to implement an automatic prefetch as successful as the experienced fileroom clerk. An algorithm based on defined metagroup categories for examination type mnemonics has been designed and implemented as one possible solution to the prefetch problem. The metagroups such as gastrointestinal (GI) tract, abdomen, chest, etc, can represent, in a small number of categories, the several hundreds of examination types performed by a typical radiology department. These metagroups can be defined in a table of examination mnemonics that maps a particular mnemonic to a metagroup or groups, and vice versa. This table is used to effect the prefetch rules of relevance. A given examination may relate to several prefetch categories, and preferences are easily configurable for a particular site. The prefetch algorithm metatable was implemented in database structured query language (SQL) using a many-to-many fetch category strategy. Algorithm performance was measured by analyzing the appropriateness of the priors fetched based on the examination type of the current study. Fetched relevant priors, missed relevant priors, fetched priors that were not relevant to the current examination, and priors not fetched that were not relevant were used to calculate sensitivity and specificity for the prefetch method. The time required for real time requesting of priors not previously prefetched was also measured. The sensitivity of the prefetch algorithm was determined to be 98.3% and the specificity 100%. Time required for on-demand requesting of priors was 9.5 minutes on average, although this time varied based on age of the prior examination and on the time of day and database traffic. A prefetch algorithm based on metatable examination mnemonic categories can pull the most appropriate relevant priors, reduce the number of missed relevant priors, and therefore reduce the time involved for the manual task of on-demand requests of priors. Network and database traffic can be reduced as well by decreasing the number of priors selected from the archive and subsequently transmitted to the display stations, through elimination of transactions on examinations not relevant to the current study. PMID- 10847368 TI - Impact of digital radiography on clinical workflow. AB - It is commonly accepted that digital radiography (DR) improves workflow and patient throughput compared with traditional film radiography or computed radiography (CR). DR eliminates the film development step and the time to acquire the image from a CR reader. In addition, the wide dynamic range of DR is such that the technologist can perform the quality-control (QC) step directly at the modality in a few seconds, rather than having to transport the newly acquired image to a centralized QC station for review. Furthermore, additional workflow efficiencies can be achieved with DR by employing tight radiology information system (RIS) integration. In the DR imaging environment, this provides for patient demographic information to be automatically downloaded from the RIS to populate the DR Digital Imaging and Communications in Medicine (DICOM) image header. To learn more about this workflow efficiency improvement, we performed a comparative study of workflow steps under three different conditions: traditional film/screen x-ray, DR without RIS integration (ie, manual entry of patient demographics), and DR with RIS integration. This study was performed at the Cleveland Clinic Foundation (Cleveland, OH) using a newly acquired amorphous silicon flat-panel DR system from Canon Medical Systems (Irvine, CA). Our data show that DR without RIS results in substantial workflow savings over traditional film/screen practice. There is an additional 30% reduction in total examination time using DR with RIS integration. PMID- 10847369 TI - Transmission failure rate for computed tomography examinations in a filmless imaging department. AB - The purpose of the study was to determine the frequency and causes of unsuccessful computed tomography (CT) transmissions in a filmless imaging department and to determine the added efficiency gains provided by the sequential addition of modality worklist software and a major network upgrade. Prospective data on CT transmission error rates were recorded over an 18-month period. During the study interval, modality worklist functionality was added, followed by a network upgrade. Failed transmissions were categorized as to the source of the error (human v technical), and the specific problem encountered. Prior to the introduction of modality worklist software, the initial CT transmission failure rate was 7.6%, which was primarily the result of human error (69%), in the form of data entry error. Upon the introduction of modality worklist software, the transmission failure rate decreased to 3.5%, with human error accounting for only 16% of all failed transmissions. The subsequent addition of a network upgrade from shared Ethernet to switched Ethernet further reduced the transmission failure rate to 2.0%, which was believed to be the result of a reduction in the number of network collisions. Other sources of failed transmission occur at the levels of the CT scanner (network interface card), picture archiving and communication system (PACS)/hospital information system (HIS) interface, and modality gateway. When planning the transmission from film-based to filmless operation, one should consider various hardware, software, and infrastructural requirements to ensure successful PACS implementation. Software upgrades, in the form of modality worklist software, serve to improve technologist productivity by minimizing data entry error. Infrastructural changes, in the form of network upgrading, ensure proper dissemination of electronic data with decreased frequency of network collisions. Collectively, these improvements lead to enhanced transmission of digital images, resulting in productivity gains within the filmless CT department. PMID- 10847370 TI - Challenges associated with interfacing computed tomography to a picture archiving and communication system at the Baltimore Veterans Affairs Medical Center--a historical perspective. AB - The interfacing of digital image acquisition modalities to the picture archiving and communication system (PACS) plays a major part in the conversion from a traditional film-based radiology practice to one that relies almost entirely on soft-copy reading. The Baltimore Veterans Affairs Medical Center (VAMC) is one of the first filmless hospitals in the world. Since 1993, it has used computed tomography (CT) scanners connected to a commercial PACS to provide digitized patient images for filmless reading. Over the years, the evolution of Digital Imaging and Communications in Medicine (DICOM) standards, advances in networking technologies, and enhancements in PACS and hospital information system (HIS) software have greatly improved this system's robustness and patient/study identification accuracy. The result has been a major increase in productivity. PMID- 10847371 TI - Modality interfacing: the impact of a relay station. AB - We evaluated the effect of a deploying a relay station on demographic discrepancies, image segmentation for routing, quality control (QC), and technologist workflow in a distributed architecture type picture archiving and communication system (PACS) environment. A currently existing PACS environment for computed tomography (CT) was evaluated before and after the implementation of a relay station for demographic error-rate and correct study routing to the workstations. Assessment of the technologists' perceptions with respect to numerous workflow factors was performed with a questionnaire. Statistical analysis was performed using a chi-square test. The demographic error rate for CT examinations was nearly abolished with relay station deployment (14.0% pre-Relay v 0.55% post-Relay, P < .001, chi2). The technologists' perception was favorable, with a substantial majority indicating that a positive impact is made on correcting demographic errors (90%), facilitating QC (67%), and ensuring proper routing (77%). A majority also felt the user interface was intuitive (93.3%) and preferred relay (90%) over film handling but that training should be provided both by didactic sessions and "hands on" time with a trainer. The times to perform tasks were favorable for the relay station (1 to 5 minutes) versus film production and handling (2 to 15 minutes). In conclusion, the relay station prospectively eliminates demographic errors, effectively segments images from the same study routing them to different workstations, and can be seamlessly integrated into the technologists' current workflow. This can be scalable and a lower cost solution as opposed to deploying dedicated PACS QC workstations. PMID- 10847372 TI - Cable modem access to picture archiving and communication system images using a web browser over the Internet. AB - This presentation describes our experiences using a web-based viewing software and a browser to view our picture archiving and communication system (PACS) images at a remote site with cable modem-internet communications. Our testing shows that using a cable modem to access our radiology webserver produces acceptable transmission speeds to remote sites. The average time-to-display (TTD) for 16 computed tomography (CT) images on the web-based intranet system in our hospital was 7 to 8 seconds. Using a cable modem and comparable equipment at a remote site, the average TTD is 16 seconds over the internet. The TTD does not significantly change during various hours of the day. Security for our hospital based PACS is provided by a firewall. Access through the firewall is accomplished using virtual private network (VPN) software, a secure ID, and encryption. We have found that this is a viable method for after-hours subspecialty radiology consultation. PMID- 10847373 TI - Radiology workflow and patient volume: effect of picture archiving and communication systems on technologists and radiologists. AB - This study was performed to evaluate the changes in workflow and efficiency in various clinical settings in the radiology department after the introduction of a picture archiving and communication system (PACS). Time and motion data were collected when conventional image management was used, and again after the introduction of a PACS. Changes in the elapsed time from examination request until the image dispatch to the radiologist, and from dispatch until report dictation, were evaluated. The relationship between patient volume and throughput was evaluated. The time from examination request until dispatch was significantly longer after the introduction of PACS for examinations taken on patients from the emergency department (ED) (pre-PACS, 20 minutes; post-PACS, 25 minutes; P < .0001), and for examinations taken on patients in the medical intensive care unit (MICU) (pre-PACS, 34 minutes; post-PACS, 42 minutes; P < .0001). The interval from image dispatch until report dictation shortened significantly after the introduction of PACS in the ED (pre-PACS, 38 minutes; post-PACS, 23 minutes; P < .0001) and in the outpatient department (OPD) (pre-PACS, 38 minutes; post-PACS, 20 minutes; P < .0001). Simple least squares regression showed a significant relationship between daily patient volume and the daily median time until report dictation (F = 43.42, P < .001). PACS slowed technologists by prolonging the quality-control procedure. Radiologist workflow was shortened or not affected. Efficiency is dependent on patient volume, and workflow improvements are due to a shift from batch to on-line reading that is enabled by the ability of PACS to route enough examinations to keep radiologists fully occupied. PMID- 10847374 TI - Integration of radiologist peer review into clinical review workstation. AB - Professional peer review of random prior radiologist's interpretations is mandated by the Joint Commission on Accreditation of Healthcare Organizations (JCAHO). The JCAHO expects documentation of 5% rate of random peer-review cases. Countless hours are spent in departments fulfilling these requirements. The integration of the peer-review process into the radiologist's interpretation workflow was expected to increase the percentage of documented peer review, yet decrease the time and effort for this documentation. radStation clinical review workstations are deployed at every reading station. When a requisition is bar coded, radStation retrieves the patient's clinical information and automatically displays the prior comparison report. If the radiologist agrees with the prior report, a single click on a "quality assurance' agree box documents the agreement. In the case of a discordance, an additional dialog box automatically appears and the radiologist enters the reason for disagreement and then submits the case as a discrepancy. The system holds the discordance for 3 to 5 working days, then notifies the original radiologist via E-mail that a prior interpretation has been submitted for peer review, lists the submitted discrepancy reason, and provides a link to display the discordant report. The peer-review database is separate from the existing radiology information system (RIS). At the end of every month, summary reports of all peer-review activity are generated automatically. Initial benchmarks of our deployed system anticipate documentation of long-term random peer-review rate at greater than 50% of interpreted cases. The system enhances the peer-review process by integrating it with the normal interpretation workflow. The time to complete peer review using radStation is less than 1 second per normal case and less than 60 seconds for a discordant case. The E-mail notification system is fully automated, eliminating the need for secretarial involvement in the data collection. This system has completely replaced a manual paper-based system. The integration of peer review directly into the radiologist's interpretation workstation greatly enhances the capability to easily exceed JCAHO standards. The overall increase in peer-review documentation should continue to improve the ability to document a consistent high quality of patient care. PMID- 10847375 TI - Do picture archiving and communication systems improve report turnaround times? AB - Radiology departments are beginning to embrace new technologies to decrease operating budgets and improve services. One of these technologies is the picture archiving and communication system (PACS). PACS, through immediate availability of images to the radiologist, promises to decrease turnaround times of reports to the clinician. The purpose of this study was to determine if this technology actually decreases the time for referring clinicians to receive reports generated by the radiologist. The time to provide a preliminary report by a resident and time to finalize this report by a board-certified radiologist was retrospectively obtained for 6,022 abdominal and pelvic computed tomography (CT) scans over two 1 year periods from March 1, 1997 to March 1, 1998 and from March 1, 1998 to March 1, 1999. During the first year, interpretation was conducted using hard-copy film and during the second using PACS. In both 1-year periods, MedSpeak voice recognition software (IBM, White Plains, NY) was employed for dictation. The average time for a preliminary report for a abdominal and pelvic CT, dictated by a resident or fellow, to be available in alphanumeric form on the hospital information system using hard-copy film was 3.73 days. The installation of a PACS system decreased this turnaround time to 0.56 days, representing an 85.0% improvement. The time to availability of final reports, ie, signed by board certified staff radiologists, was 5.49 days in the hard-copy interpretation subset and 5.97 days in the PACS subset. The addition of PACS into an academic gastrointestinal radiology division improves availability of alphanumeric preliminary reports of abdominal and pelvic CTs on the hospital information system (HIS), dictated by a resident or fellow, by 85.0%. There was no impact with a PACS on the time to final sign reports by a staff board certified radiologist as signing patterns remained relatively constant over the two interpretation formats. PMID- 10847376 TI - Adoption of alternative financing strategies to increase the diffusion of picture archiving and communication systems into the radiology marketplace. AB - The objective of the study was to evaluate current marketplace conditions and strategies employed by major picture archiving and communication systems (PACS) vendors in the creation of alternative financing strategies, to enhance the diffusion of filmless imaging. Data were collected from the major PACS vendors in the forms of survey questionnaires and review of existing leases. Topics evaluated in the survey included current financing options available, foreseeable changes in PACS financing, role of third-party financiers, and creation of risk sharing arrangements. Generic leases were also reviewed evaluating the presence or absence of several key variables including technology obsolescence protection, hardware/software upgrades, end-of-term options, determination of fair market value, functionality/acceptance testing, uptime guarantees, and workflow management consulting. Eight of the 10 PACS vendors surveyed participated in the data collection. The vast majority of current PACS implementations (60% to 90%) occur through direct purchase, with conventional leasing (operating or capital) accounting for only 5% to 30% of PACS installations. The majority of respondents view fee-for-lease arrangements and other forms of risk sharing as increasing importance for future PACS financing. The specific targets for such risk-sharing arrangements consist of small hospital and privately owned imaging centers. Leases currently offered range in duration from 3 to 5 years and frequently offer technology obsolescence protection with upgrades, multiple end-of-term options, and some form of acceptance testing. A number of important variables frequently omitted from leases include uptime guarantees, flexibility in changing financing or vendors, and incorporation of expected productivity/operational efficiency gains. As vendors strive to increase the penetration of PACS into the radiology marketplace, there will be a shift from conventional financing (loan or purchase) to leasing. Fee-for-use leasing and other forms of risk sharing have the greatest potential in smaller hospitals, which do not have the financial resources to pursue conventional financing options. Potential PACS customers must be cautious when entering into these alternative financing strategies, to ensure that appropriate safeguards are incorporated, in order to minimize downside risk. PMID- 10847377 TI - Using technology assessment as the picture archiving and communication system spreads outside radiology to the enterprise. AB - Picture archiving and communication systems (PACS) are being implemented within radiology departments, and many facilities are entering the next stage of PACS use by deploying PACS to departments outside of radiology and to other facilities located at a distance. Many PACS vendors and department administrators have based cost-justification analyses on the anticipated savings from expanding PACS to these areas. However, many of these cost-savings analyses can be highly suspect in their assumptions and findings. Technology assessment (TA) at the hospital/health system level is an organized, systematic approach to examining the efficacy of a technology in relation to the health system's mission and clinical needs. It can be an organized and unifying approach to aid in the distribution of limited capital resources. As extra-radiology PACS deployment is a costly endeavor, TA may be used to plan for PACS implementation throughout the enterprise. In many organizations, PACS is thought of as a radiology domain as its first uses were centered on this image-producing service. Now, as PACS technology spreads to other service areas, such as cardiology, dermatology, pathology, orthopedics, obstetrics, etc, the need to incorporate other viewpoints in a system-based PACS is necessary to avoid having independent PACS that may duplicate archives and may not communicate with each other. How to meet the diverse PACS needs of clinical services can be a challenging task; a TA program has been demonstrated to effectively handle the clinical needs, demands, and timeframes of PACS planning and support throughout hospitals and health systems. A hospital-based TA program can assist health care organizations to present PACS as a system-wide need and program rather than a radiology-based program gobbling up the capital budget. Submitting PACS to the TA review process can identify essential elements in planning and help avoid many of the pitfalls of PACS implementation and operations. Thorough cost and/or return on investment analyses, phasing decisions, workflow re-engineering, and outcomes assessment programs are a few of the issues that a TA program can address to help in the transition to a complete electronic image environment. The TA process includes clinician selection, evaluation criteria and their selection for technologies under review, a policy for review/authorization/denial, and measurement of expected outcomes. PMID- 10847378 TI - A new needle-crystalline computed radiography detector. AB - The most successful digital radiography detectors to date have been storage phosphor plates used in computed radiography (CR). The detector is cheap, has good producibility, and is robust. Direct radiography (DR) systems are being developed based on flat-panel technology. Better image quality is claimed for some DR systems. On the other hand, DR detectors have low producibility and robustness, and a high price. A new CR detector is being developed at Agfa that combines the advantages of CR and DR. It is a storage phosphor plate made up of needle-shaped crystals. The phosphor efficiently converts absorbed x-ray quanta into photostimulable centers for efficient read out. It has a large dynamic range and its emission is efficiently detected with both photomultiplier tube (PMT) and charge coupled device (CCD). It is shown that CR systems based on the new detector offer image quality that matches that of the best DR systems. PMID- 10847379 TI - Quantitative methods in phase-contrast x-ray imaging. AB - A new method for extracting quantitative information from phase-contrast x-ray images obtained with microfocus x-ray sources is presented. The proposed technique allows rapid noninvasive characterization of the internal structure of thick optically opaque organic samples. The method does not generally involve any sample preparation and does not need any x-ray optical elements (such as monochromators, zone plates, or interferometers). As a consequence, samples can be imaged in vivo or in vitro, and the images are free from optical aberrations. While alternative techniques of x-ray phase-contrast imaging usually require expensive synchrotron radiation sources, our method can be implemented with conventional, albeit microfocus, x-ray tubes, which greatly enhances its practicality. In the present work, we develop the theoretical framework, perform numerical simulations, and present the first experimental results, demonstrating the viability of the proposed approach. We believe that this method should find wide-ranging applications in clinical radiology and medical research. PMID- 10847380 TI - Extraction of microcalcifications in digital mammograms using regional watershed. AB - In this report, a novel technique is proposed for computer-aided automatic extraction of microcalcifications in a digital mammogram. First, the microcalcifications are detected by morphological filtering, followed by entropy based thresholding. Next, the microcalcifications are segmented by computing regional watershed. The proposed automatic technique is designed to serve as a visual aid to radiologists. Its efficacy is demonstrated through experimental results. PMID- 10847381 TI - A multiscale algorithm for segmenting calcifications from high-resolution mammographic specimen radiographs. AB - We have developed a multiscale algorithm for segmenting breast calcifications from high-resolution specimen radiographs. The algorithm was evaluated using 152 mammographic regions of interest digitized at a 15-microm spatial resolution. The true-positive detection rate was approximately 97.4% with 0.67 false-positives per image, and the segmentation error of individual calcification particles was approximately 5%. The performance of the algorithm is highly satisfactory. PMID- 10847382 TI - The use of digital imaging and communications in medicine (DICOM) in the integration of imaging into the electronic patient record at the Department of Veterans Affairs. AB - The US Department of Veterans Affairs (VA) is using the Digital Imaging and Communications in Medicine (DICOM) standard to integrate image data objects from multiple systems for use across the health care enterprise. DICOM uses a structured representation of image data and a communication mechanism that allows the VA to easily acquire images from multiple sources and store them directly into the online patient record. The VA can obtain both radiology and nonradiology images using DICOM, and can display them on low-cost clinician's color workstations throughout the medical center. High-resolution gray-scale diagnostic quality multimonitor workstations with specialized viewing software can be used for reading radiology images. The VA's DICOM capabilities can interface six different commercial picture archiving and communication systems (PACS) and more than 20 different image acquisition modalities. The VA is advancing its use of DICOM beyond radiology. New color imaging applications for gastrointestinal endoscopy and ophthalmology using DICOM are under development. These are the first DICOM offerings for the vendors, who are planning to support the recently passed DICOM Visible Light and Structured Reporting service classes. Implementing these in VistA is a challenge because of the different workflow and software support for these disciplines within the VA hospital information system (HIS) environment. PMID- 10847383 TI - Can radiologic images be incorporated into the electronic patient record? AB - As radiology makes advances toward filmlessness, all of medicine is headed, just as rapidly, toward paperless transmission of patient information. While there are obvious advantages to this electronic approach, and several standards to conform to for the transmission of textual (Health Level 7 [HL-7]) and image (Digital Imaging and Communications in Medicine [DICOM]) data, it is the integration of these two data sets that is clinically essential and yet poorly defined. This report defines an approach for, and the successful implementation of, the integration of radiologic image data with textual data contained within the electronic patient record (EPR) through the use of standard internet protocols. Incorporation of medical images in the EPR has proven to be critical to the successful deployment of picture archiving and communications systems (PACS) and the reduction of film consumption at Massachusetts General Hospital (MGH). Since the installation of the first internet-based Image Data Repository (IDR) at MGH in 1995, the system has adequately served to meet the needs of clinical requests by both radiology-only browser users and users of the EPR. It has drastically reduced the need for film and provided concurrent display of images and text throughout the institution and beyond. PMID- 10847384 TI - Implementing a Java-based image and report distribution system in a non-picture archiving and communication system environment. AB - The benefits and pitfalls of implementing a Java-based system to distribute results and images to referring physicians are addressed. The basic requirements for and barriers to implementing this system in a non-picture archiving and communication system (PACS) environment will also be discussed. The majority of radiology information systems (RIS) and hospital information systems (HIS) currently only distribute the text data for radiology examinations. This is generally adequate for low-acuity exams in a relatively healthy patient; however, many clinicians prefer to review images so they can correlate the reported findings with the image data, as well as review the exam themselves. A web-based solution eliminates the need for specialized review software and/or hardware at each review site. In addition, there is no need for support personnel to travel to each site to set up and upgrade software. PMID- 10847385 TI - The web browser as a unifying agent for radiology and the health care enterprise. AB - A variety of information systems in the health care enterprise are used to store patient information. The hospital information system (HIS), the picture archiving and communications system (PACS), the radiology information systems (RIS), and patient records, for example, are often supported by separate and distinct systems. A referring physician reviewing a radiologist's report often does not have convenient access to the original radiologic images. A radiologist interpreting a radiologic examination may not have convenient access to clinical information generated outside of the radiology department. Clearly, convenient and quick access to all relevant patient information is the ideal in delivering quality patient care. Recently, the web browser has become increasingly well accepted as the "universal client" for viewing a wide variety of information. We believe that the web browser may provide the universal "window" to the variety of distinct information sources required by the physician, and may provide unification of these information sources through hypertext links. We specifically looked at the feasibility of using the web browser to view scanned patient charts. A total of 5,200 patient charts (131,000 pages) were scanned into a structured query language (SQL) database. Access to these records was made available on the hospital local area network (LAN), and on the Internet over the worldwide web. Comparison measurements were made of the time required to access patient records using a dedicated viewing application and using a web browser. PMID- 10847386 TI - Performance and function of a desktop viewer at Mayo Clinic Scottsdale. AB - A clinical viewing system was integrated with the Mayo Clinic Scottsdale picture archiving and communication system (PACS) for providing images and the report as part of the electronic medical record (EMR). Key attributes of the viewer include a single user log-on, an integrated patient centric EMR image access for all ordered examinations, prefetching of the most recent prior examination of the same modality, and the ability to provide comparison of current and past exams at the same time on the display. Other functions included preset windows, measurement tools, and multiformat display. Images for the prior 12 months are stored on the clinical server and are viewable in less than a second. Images available on the desktop include all computed radiography (CR), chest, magnetic resonance images (MRI), computed tomography (CT), ultrasound (U/S), nuclear, angiographic, gastrointestinal (GI) digital spots, and portable C-arm digital spots. Ad hoc queries of examinations from PACS are possible for those patients whose image may not be on the clinical server, but whose images reside on the PACS archive (10TB). Clinician satisfaction was reported to be high, especially for those staff heavily dependent on timely access to images, as well as those having heavy film usage. The desktop viewer is used for resident access to images. It is also useful for teaching conferences with large-screen projection without film. We report on the measurements of functionality, reliability, and speed of image display with this application. PMID- 10847387 TI - The implementation of speech recognition in an electronic radiology practice. AB - For both efficiency and economic reasons, our practice (200,000 examinations) has converted all remote dictation to speech recognition transcription (PowerScribe, L & H, Burlington, MA). The design criteria included complete automation to the existing radiology information system (RIS), with full RIS capabilities immediately available following dictation. All dictations for computed tomography, magnetic resonance imaging, ultrasound, and nuclear medicine were converted from remote transcription to speech recognition over a 2-week period (following a 4-week installation phase and 8 days of training). The average turnaround time for these reports decreased from approximately 2 hours to less than 1 minute. Reports are then sent to the institutional Electronic Medical Record and are available throughout all facilities in a nominal 2 minutes. Speech recognition rates were surprisingly high, although certain phrases caused consistent difficulties and certain staff required retraining. This presents our analysis of both successful and problematic areas during our design and implementation, as well as statistical performance analyses. PMID- 10847388 TI - Active matrix liquid crystal displays for clinical imaging: comparison with cathode ray tube displays. AB - Fifteen large-area, flat-panel displays used for clinical image review were evaluated for image quality and compared with 30 comparably sized cathode ray tube (CRT) monitors. Measurements were of image display patterns by Video Electronic Standards Association (VESA) and a commercial product. Field measurements were made of: maximum and minimum luminance, ambient lighting, characteristic curve (gamma), point shape and size, high-contrast resolution, uniformity, and distortion. Assessments were made of pixel defects, latent image patterns, ghosting artifacts, and viewing angle luminance. Also, a questionnaire was generated for users of the flat-panel and CRT units. Seventeen respondents indicated no preference for either flat panel or CRT. Results show these flat panels to have higher luminance (mean, 177.7 cd/m2); larger number of just noticeable differences (JNDs; n = 555), higher gamma, comparable uniformity, and warm-up time. CRTs had less angle viewing dependence and far fewer artifacts (ghosting and latent images). Our questionnaire showed active matrix liquid crystal displays (AMLCD) to be fully acceptable for clinical image viewing. Furthermore, the statistical results show that further testing for new AMLCDs of this type is unwarranted. PMID- 10847389 TI - Contrast-detail characteristic evaluations of several display devices. AB - The contrast-detail characteristic of a display system is a powerful tool for evaluating displayed image quality. It takes into account the physical properties of the display, the psychophysical aspects of the observer, and the viewing conditions. It is a more sensitive measurement of the displayed image quality than a simple Society of Motion Picture and Television (SMPTE) pattern. Yet, it is relatively simple to measure and requires no special equipment or analysis tools. In this presentation, the results of the evaluation of several cathode ray tube (CRT) monitors and a digital projector will be presented. Contrast-detail characteristics of these display devices were measured under various gamma and display settings. The results show excellent intraobserver and interobserver variance (<1 step on the grayscale). Extraneous light, such as room lighting, affects the contrast threshold more severely at low background levels more than at high background. Gamma settings on graphics adapters affect the shapes of the contrast-detail curve for all display types. Gamma settings of approximately 2.0 result in a better contrast threshold for both high and low background brightness. The results show complex differences in contrast-detail characteristics for different display types. The digital projector display not only has significantly worse performance than CRT monitors, but also is affected more by extraneous light. High-brightness monitors with optimal monitor and graphics adapter settings have better performance than color or low-brightness monitors. However, under some settings, the performance of high-brightness monitors is not always better at all object sizes and background levels. PMID- 10847390 TI - Simulation of disaster recovery of a picture archiving and communications system using off-site hierarchal storage management. AB - The purpose of this communication is to report on the testing of the disaster recovery capability of our hierarchical storage management (HSM) system. Disaster recovery implementation is a requirement of every mission-critical information technology project. Picture archiving and communications systems (PACS) certainly falls into this category, even though the counterpart, conventional film archive, has no protection against fire, for example. We have implemented a method for hierarchical storage with wavelet technology that maximizes on-site case storage (using lossy compression), retains bit-preserved image data for legal purposes, provides an off-site backup (lossless bit-preserving wavelet transform), and provides for disaster recovery. Recovery from a natural (earthquake and subsequent fire) or technical (system crash and data loss) disaster was simulated by attempting to restore from the off-site image and database backup to clean core PACS components. The only existing loaded software was the operating system. The database application was reloaded locally, and then the database contents and image store were loaded from the off-site component of the HSM system. The following measurements were analyzed: (1) the ability to recover all data; (2) the integrity of the recovered database and image data; (3) the time to recover the database relative to the number of studies and age of the archive, as well as bandwidth between the local and remote site; and (4) the time to recover image data relative to compression ratio, number of studies, number of images, and time depth of the archive. This HSM system, which maximizes on-site storage, maintains a legal record, and provides off-site backup, also facilitates disaster recovery for a PACS. PMID- 10847391 TI - "Permanent" records: experience with data migration in radiology information system and picture archiving and communication system replacement. AB - In the replacement of both a radiology information system (RIS) and a picture archiving and communication system (PACS) archive, data were migrated from the prior system to the new system. We report on the process, the time and resources required, and the fidelity of data transfer. We find that for two PACS archives, both organized according to the Digital Imaging and Communications in Medicine (DICOM) information model, data may be transferred with full fidelity, but the time required for transfer is significant. Transfer from off-line backup media was found to be faster than transfer from our robotic tape library. In contrast, the RIS replacement required extensive labor to translate prior data between dissimilar information models, and some data were inevitably lost in the translation. Standards for RIS information models are needed to promote the migration of data without loss of content. PMID- 10847392 TI - Demystifying the hospital information system/radiology information system integration process. AB - Most organizations planning to implement picture archiving and communications systems (PACS) are aware of the need to integrate the hospital information system (HIS) and radiology information system (RIS) with the PACS, yet few are acutely aware of the challenges associated with this requirement. This report highlights the results of collaborative efforts between Children's Hospital Medical Center Cincinnati (CHMC) applications specialists with expertise in the HIS and CHMC information system, radiology staff familiar with the enterprise and radiology workflow and data flow requirements; and General Electric integration engineers familiar with the SMS HIS and RIS, and GE PACS. CHMC received Board approval, including full funding of the entire PACS project, in October 1998. An aggressive time frame for installation was established, as CHMC's PACS leadership committed to the selection, design, and implementation of PACS and computed radiography (CR) within 18 to 20 months. CHMC selected GE (Milwaukee, WI) as its PACS vendor in July 1999, and began its implementation in November 1999. We will present the four-stage integration process undertaken at CHMC: (1) planning the integration effort, (2) designing the Interface, (3) building the interface, and (4) testing the Interface. PMID- 10847393 TI - Upgrading legacy systems for the Integrating the Healthcare Enterprise (IHE) initiative. AB - As technology vendors have adopted standardized communication protocols, including Digital Imaging and Communications in Medicine (DICOM) and Health Level 7 (HL7), interconnectivity between various devices has been simplified. The recent Integrating the Healthcare Enterprise (IHE) initiative will further promote the use of standards for interconnectivity. Until these standards are universally accepted, we must live in a transitional world where some components will communicate without any modification, while others require upgrades to allow them to meet the new standards. In designing and implementing the network at University of California Los Angeles (UCLA) Medical Center, some integration problems were found that are common to the industry. Creating departmental workflow with only a limited number of acquisition devices supporting the DICOM worklist was the initial problem addressed. Although many manufacturers provide this function for their new scanners, upgrading existing equipment is often cost prohibitive. To ensure the quality of the demographic information in the image data and the workflow of the system, third-party worklist components were required to upgrade the legacy acquisition devices. These worklist components provided a standards-compliant facade on top of the legacy equipment, allowing seamless integration with the remainder of the network. To support the distribution of worklist information and the feedback of procedure status, a bidirectional HL7/DICOM protocol bridge was required. Although many radiology information system (RIS) manufacturers will be providing native DICOM capabilities in future product releases, the majority of current RIS installations have no DICOM functionality. Similar to the legacy scanners, the HL7/DICOM bridge provided a DICOM-compliant facade to the non-DICOM RIS. The additional use of web-based technology for worklist display further extended flexibility of this facade. We have demonstrated standards-compliant facade technology allowing legacy components to operate seamlessly in an IHE environment at a fraction of the cost of upgrading to new equipment. PMID- 10847394 TI - An evaluation of JPEG and JPEG 2000 irreversible compression algorithms applied to neurologic computed tomography and magnetic resonance images. Joint Photographic Experts Group. AB - We performed visual comparison of 200 head magnetic resonance (MR) and 200 head computed tomography (CT) images compressed at two levels using standard Joint Photographic Experts Group (JPEG) irreversible compression and a preliminary version of the JPEG 2000 irreversible algorithm. Blinded evaluations by neuroradiologists compared original versus either JPEG or JPEG 2000. We found that this version of JPEG 2000 did not perform as well as the current JPEG for head CTs, but for MR images, JPEG 2000 performed as well or better. Around 7:1 compression ratio seemed to be a conservative point where there was no perceptible difference. PMID- 10847395 TI - An investigation of the effects of mammographic acquisition parameters on a semiautomated quantitative measure of breast cancer risk. AB - The aim of this work was to investigate the effect of mammographic acquisition parameter variations on the estimation of percent density (PD) produced by a particular semiautomated algorithm. The PD algorithm requires the user to specify a threshold pixel value segmenting breast tissue of greater and lesser density. A whole breast specimen was imaged using a variety of acquisition techniques, and the image data were processed as prescribed by the PD algorithm. PD estimates for all possible values of the user threshold were calculated for all the images. The image data were normalized so that PD varied between 30% and 80% over a fixed threshold range of 23, and a PD value of 50% was obtained for a threshold value of 195. PD differences between all the images and a baseline standard mammographic acquisition technique were calculated. We also estimated PD differences caused by small (3%) variations in operator selection of the threshold value. We found that the largest differences in PD involved changes in the density control of the mammography unit, and changes in the detector (either film type or computed radiography). The maximum PD differences due to technique were all less than 10%, with root-mean-square (RMS) variations less than 4%. PD differences due to operator variation were 24% (maximum) and 6.1% (RMS). These findings suggest that PD differences due to mammographic technique will be considerably less than those inherent to the technique, due to operator variation. All of these estimates are likely larger than differences seen in practice since optimization of the threshold by the operator was not considered in this analysis. PMID- 10847396 TI - Analyzing the impact of wavelet compression on radioisotope image quality. PMID- 10847397 TI - Clinical evaluation of irreversible compression of ultrasound images using the JPEG algorithm at approximately 9:1. Joint Photographic Expert's Group. PMID- 10847398 TI - Lesion characterization using vessel permeability map to new blood pool contrast agent calculated from dynamic magnetic resonance images. AB - Use of contrast enhancement in conjunction with magnetic resonance (MR) imaging provides a means to evaluate tissue function, as well as morphology. Moreover, physiologic properties derived from kinetic analysis of dynamic contrast-enhanced data can improve the specificity of MR examinations. In this study, quantitative analysis of microvascular characteristics based on dynamic MR imaging were performed both for malignant and benign lesions using two types of contrast agents (CAs). A new MR macromolecular contrast medium (MMCM), 24 gadolinium tetraazacyclododecanetetraacetic acid (DOTA)-dendrimer, was found to have a greater ability to distinguish benign from malignant lesions. When a blood pool agent was used, permeability differences in the two types of lesions were the most significant findings among all parameters considered. PMID- 10847399 TI - Coregistration of brain single-positron emission computed tomography and magnetic resonance images using anatomical features. AB - The present report describes a computer tool for the coregistration of single positron emission computed tomography (SPECT) and magnetic resonance (MR) images to aid the diagnosis. Two types of images are used for some neurologic diseases: one of them anatomic (magnetic resonance) and the other metabolic (SPECT), with the specialist being required to make a mental integration of the examinations. This procedure can be improved by using a computer tool that might permit the presentation of this information in a single image. The coregistration is performed on the basis of pairs of points positioned by a specialist according to the structures present in the images and the least squares error is calculated between them using Euclidean distance. Coinciding planes and section thickness are selected for the two modalities and the SPECT image is processed so as to have the same spatial resolution as the resonance image. PMID- 10847400 TI - Impact of digital radiography on clinical workflow and patient satisfaction. AB - Compared to traditional film radiography, digital radiography is believed to improve workflow and patient throughput. Digital radiography permits the technologist to immediately view the quality of the film directly at the modality. Additional workflow improvements, therefore, should be achieved with the integration of the radiology information system (RIS). To learn more about this proposed efficacy, a study was performed at The Cleveland Clinic Foundation (Cleveland, OH) comparing timings in three groups: traditional film radiography; digital radiography; and digital radiography with RIS integration. Our data validated a timesaving of digital radiography over traditional or standard films and an even greater timesaving in a digital radiography/RIS environment. PMID- 10847401 TI - Security model for picture archiving and communication systems. AB - The modern information revolution has facilitated a metamorphosis of health care delivery wrought with the challenges of securing patient sensitive data. To accommodate this reality, Congress passed the Health Insurance Portability and Accountability Act (HIPAA). While final guidance has not fully been resolved at this time, it is up to the health care community to develop and implement comprehensive security strategies founded on procedural, hardware and software solutions in preparation for future controls. The Virtual Radiology Environment (VRE) Project, a landmark US Army picture archiving and communications system (PACS) implemented across 10 geographically dispersed medical facilities, has addressed that challenge by planning for the secure transmission of medical images and reports over their local (LAN) and wide area network (WAN) infrastructure. Their model, which is transferable to general PACS implementations, encompasses a strategy of application risk and dataflow identification, data auditing, security policy definition, and procedural controls. When combined with hardware and software solutions that are both non performance limiting and scalable, the comprehensive approach will not only sufficiently address the current security requirements, but also accommodate the natural evolution of the enterprise security model. PMID- 10847402 TI - Effects of a large-scale deployment of soft-copy devices for picture archiving and communication systems viewing. AB - A study was developed to investigate several aspects of deployment of stand-alone computers and a web-based server for displaying digital images. The costs associated with installation, upgrades, training, and maintenance of both systems were documented, as well as patterns of usage and preferences of physicians for the two types of systems. A clinical archive was created to store relevant images with a web-based front end. Users were classified according to their imaging requirements and were given access to images from either the diagnostic archive (full study sets) or the clinical archive. The range of functionality of the web based system was significantly lower than that of the stand-alone system; however, the costs associated with the administration of the stand-alone systems were far higher than those associated with a web server. A clinical archive was created to store relevant images with a web-based front end. This study illustrates the different approaches to distribution of images to clinicians with the costs of administration of each system. PMID- 10847403 TI - Electronic imaging system implementation at Mayo Clinic Rochester: downtime procedures and communication plans. AB - Electronic imaging system implementation is a challenging process and requires: (1) thorough procedure documentation and communication plans; (2) systematic evaluation of processes by all involved personnel groups; and (3) anticipation of potential issues and documentation of new problems as they occur. Our department is continuously improving procedures and communications as opportunities arise. A downtime procedure manual and a basic communication plan has given us a foundation to build upon and will help us to maintain appropriate levels of patient care throughout electronic system failures. PMID- 10847404 TI - Picture archiving and communication systems project management using web-based tools. AB - As the technology of picture archiving and communications systems (PACS) improves and implementation becomes more widespread, the project management of deploying substantially large, multiple-facility systems becomes an integral part of success. A successful deployment requires project support from the initial planning and surveying to the final acceptance, even encompassing support during active use of the PACS. The sharing of information between project stakeholders of a PACS implementation can be daunting at times, but with the flexibility of the worldwide web, this aspect can be eased. This report speaks to the tools and usability of the worldwide web to disseminate project management information for planning, implementation, and support of any PACS implementation--anywhere. This sharing of knowledge prepares the end user for what will be available for them when the complete systems is in place, allowing for a smoother migration to PACS. PMID- 10847405 TI - Evaluation of the accuracy of a continuous speech recognition software system in radiology. PMID- 10847406 TI - Implementation and day-to-day usage of a client-server-based radiology information system. AB - Implementation of a second-generation radiology information system (RIS) requires attention to many issues, including work flow, system design, training, consideration for emerging technologies, and support. This presentation covers the issues, and solutions, involved in implementation and subsequent day-to-day usage of a client-server-based RIS tightly integrated with a hospital information system (HIS) using "thin client" software to limit hardware requirements for the client computers. The software and hardware implementation had to be designed for potential enterprise-wide scalability of the RIS for a system involving 11 hospitals. Issues arose regarding initial increased turnaround times, errors in importation of historical data, and problems with implementation of interfaces between the RIS and the billing system and between the RIS and the digital dictation system. Network errors and difficulties in using a thin client implementation had to be overcome. Hierarchical training was implemented, as support was switched from an "on-site" support group to a central Information System Division with responsibility to the entire enterprise. PMID- 10847407 TI - Medical image transmission on low-speed networks. PMID- 10847408 TI - Magnetic resonance imaging deformation-based segmentation of the hippocampus in patients with mesial temporal sclerosis and temporal lobe epilepsy. AB - We compared manual and automated segmentations of the hippocampus in patients with mesial temporal sclerosis. This comparison showed good precision of the deformation-based automated segmentations. PMID- 10847409 TI - Vesselize: a system for analysis and visualization of magnetic resonance angiography. PMID- 10847410 TI - Detection of microcalcifications by means of multiscale methods and statistical techniques. AB - The detection of clustered microcalcifications can help the radiologist to detect early breast cancer. Microcalcifications exhibit some important characteristics, such as small size and high luminosity. Use of a computer-aided diagnosis (CAD) method can prevent them being overlooked. In this report, a multiresolution analysis is performed based on a multilevel wavelet transformation. Decomposition produces sub-band images which become visible only as details of the different scales. Thereafter, all the images will be combined in a final image, in order to obtain an image that contains all the interest details at the scale where microcalcifications tend to appear. Once the image, called detail image, is obtained, it is necessary to determine which details correspond with microcalcifications. Statistical analysis of the histogram permits classification of the zones likely to contain microcalcifications. Applying this statistical techniques over the whole image and representing the results in a two-dimensional map, clustered microcalcification regions are clearly distinguishable. PMID- 10847411 TI - Optimization of viewing conditions and phantom image quality evaluations on GE DMR and full-field digital mammography system. PMID- 10847412 TI - Planning factors for developing an enterprise-wide picture archiving and communication system maintenance program. AB - Picture archiving and communication system (PACS) maintenance on an individual site basis has historically been a complex and costly challenge. With the advent of enterprise-wide PACS projects such as the Virtual Radiology Environment (VRE) project, the challenge of a maintenance program with even more complexities has presented itself. The approach of the project management team for the VRE project is not one of reactive maintenance, but one of highly proactive planning and negotiations, in hopes of capitalizing on the economies of scale of an enterprise wide PACS maintenance program. A proactive maintenance program is one aspect of life-cycle management. As with any capital acquisition, life-cycle management may be used to manage the specific project aspects related to PACS. The purpose of an enterprise-wide warranty and maintenance life-cycle management approach is to maintain PACS at its maximum operational efficiency and utilization levels through a flexible, shared, yet symbiotic relationship between local, regional, and vendor resources. These goals include providing maximum operational performance levels on a local, regional, and enterprise basis, while maintaining acceptable costs and resource utilization levels. This goal must be achieved without negatively impacting point of care activities, regardless of changes to the clinical business environment. PMID- 10847413 TI - Reconstruction of electroncephalogram brain maps by incorporating the blind source separation concept. AB - Electroencephalogram (EEG) brain maps provide useful and reliable neurodiagnostic information. Accurate reconstruction of the data requires an efficient separation of the electrode signals. Although autoregressive (AR) spectrum estimation highly refines the signals, it cannot remove the effect of adjacent electrode signals. This report describes an efficient blind signal separation (BSS) method. The algorithm identifies the coefficients of an adaptive FIR filter by minimization of a cost function in terms of the corresponding fourth-order cumulants. Applying this method, the quality of the results is far superior to the traditional methods. PMID- 10847414 TI - Secure, web-accessible call rosters for academic radiology departments. AB - Traditionally, radiology department call rosters have been posted via paper and bulletin boards. Frequently, changes to these lists are made by multiple people independently, but often not synchronized, resulting in confusion among the house staff and technical staff as to who is on call and when. In addition, multiple and disparate copies exist in different sections of the department, and changes made would not be propagated to all the schedules. To eliminate such difficulties, a paperless call scheduling application was developed. Our call scheduling program allowed Java-enabled web access to a database by designated personnel from each radiology section who have privileges to make the necessary changes. Once a person made a change, everyone accessing the database would see the modification. This eliminates the chaos resulting from people swapping shifts at the last minute and not having the time to record or broadcast the change. Furthermore, all changes to the database were logged. Users are given a log-in name and password and can only edit their section; however, all personnel have access to all sections' schedules. Our applet was written in Java 2 using the latest technology in database access. We access our Interbase database through the DataExpress and DB Swing (Borland, Scotts Valley, CA) components. The result is secure access to the call rosters via the web. There are many advantages to the web-enabled access, mainly the ability for people to make changes and have the changes recorded and propagated in a single virtual location and available to all who need to know. PMID- 10847415 TI - Starting to think digitally: evaluation of web browser-based presentation of images at a Rad Path conference in a traditional film-based environment. AB - Images have traditionally been projected from film to a large-screen television at the weekly Rad Path conference at University of Medicine and Dentistry of New Jersey-New Jersey Medical School. An alternative system was developed to digitally transfer computed tomography (CT) scans and magnetic resonance images (MRI) into a standard PC format, and present them at the conference in a hypertext markup language (HTML) document via a laptop computer and liquid crystal display (LCD) projector. Resident and attending physicians were given a questionnaire about the differences between the two modes of presentation. Their responses are presented here. PMID- 10847416 TI - Virtual anatomy and movement of lower extremities using virtual reality modeling language. AB - In medical imaging, the use of three-dimensional image is increasing for both educational and diagnostic purposes. With the advent of techniques for browsing a three-dimensional object, it became possible to display three-dimensional images on a personal computer via the Internet. This report describes the construction of a three-dimensional virtual model of human lower extremities that was linked with the data acquired from gait analysis in order to perform three-dimensional gait analysis. PMID- 10847417 TI - Reduced platelet serotonin content in rabbits with dietary hypercholesterolemia. AB - Serotonin [5-hydroxytryptamine (5-HT)] has been implicated in platelet activation and vasoconstriction, two processes that contribute to arterial thrombosis in atherosclerotic diseases. In the present study, Japanese White rabbits fed 1% cholesterol for 5 weeks were used to investigate the response of hypercholesterolemic vascular arteries and platelets to 5-HT. Contractions of the thoracic aorta induced by 5-HT were comparable between the cholesterol-fed group and the age-matched control group. However, acetylcholine-induced vasodilation in arteries preconstricted with 5-HT was moderately but significantly attenuated in the cholesterol-fed rabbits. Platelet aggregation responses to 5-HT (0.1-3 micromol/l) in combination with epinephrine (5 micromol/l), adenosine diphosphate (ADP) (0.3-10 micromol/l), 9,11-dideoxy-9alpha,11alpha-methanoepoxy-prostaglandin F2alpha (U-46619) (1-30 micromol/l) or collagen (3 microg/ml) were significantly enhanced in cholesterol-fed rabbits. In contrast, platelet 5-HT content determined with a high-performance liquid chromatography-electrochemical detector (HPLC-ECD) was significantly decreased in cholesterol-fed rabbits. These results suggest a possible association among the endothelial dysfunction, platelet aggregation and platelet 5-HT content in rabbits with dietary hypercholesterolemia. PMID- 10847418 TI - The A20210 allele in the prothrombin gene enhances the risk of venous thrombosis in carriers of inherited protein S deficiency. AB - Sixteen families with inherited protein S deficiency and venous thromboembolism (VT) were screened for the presence of factor V (FV) Leiden mutation and for the G20210A allele in the prothrombin gene. While FV Leiden was not detected in any of the families, protein S deficiency and prothrombin mutation were present in five families. To assess the risk of VT in carriers of the combined defects, a total of 92 members of the 16 families, including propositi, were examined. Thirty subjects were normal, 40 showed protein S deficiency, 10 the prothrombin mutation and 12 showed both abnormalities. When index cases were excluded, thrombosis history were present in 40.7% of protein S-deficient patients, 75% of patients with combined abnormality, one out of the 10 (10%) with prothrombin mutation and only one (3.3%) of the normal subjects. Relatives with combined defects showed the highest incidence rate of VT in comparison with normal relatives (rate ratio = 32.4), those with protein S deficiency an intermediate degree (rate ratio = 15.7), and G20210A relatives the lowest (rate ratio = 3.4). Relatives with combined defects had an increased risk of VT in comparison with relatives with protein S deficiency (incidence rate ratio 2.1; 95% confidence interval, 0.7-5.41; P = 0.1). In conclusion, the presence of the prothrombin mutation seems to increase the risk of VT carriers of protein S deficiency, although additional families are required to fully estimate the magnitude of risk. PMID- 10847419 TI - A comparative study of functional assays for tissue factor pathway inhibitor using normal plasma and clinical samples. AB - Tissue factor pathway inhibitor (TFPI) is a Kunitz-type inhibitor that regulates the initiation of tissue factor-mediated coagulation. Recent reports in the literature have described variable results using different methodologies for TFPI measurement. In this study, we used one clotting and two amidolytic methodologies to assess TFPI functional levels. The study groups included normal healthy donors as well as patients with acute hepatitis, diabetes, coronary artery bypass graft operations, deep vein thrombosis, and prior to and during heparin therapy. The aims were to compare the results obtained in normal plasma using different assay systems, to compare TFPI levels in a range of clinical samples, including those previously not determined using a clotting methodology, and to report TFPI levels in patient groups previously not investigated. The results clearly demonstrate poor correlation between functional TFPI values using the different methodologies, highlighting the requirement for standardization. PMID- 10847420 TI - More porous fibrin gel structure obtained by interaction with Lys-plasminogen than with Glu-plasminogen. AB - The effect of Glu1- and Lys78-plasminogen on the assembly and structure of fibrin gels was studied in purified fibrinogen-thrombin system and in plasminogen-free plasma, using turbidity, liquid permeation and three-dimensional (3D) confocal laser microscopy methods. In the purified fibrinogen system using the turbidity method, the final optical density of the fibrin gels increased with increasing concentrations of Lys-plasminogen. The fiber mass/length ratio mu increased with increasing concentrations of both Glu1- and Lys78-plasminogen, the effect of Lys78-plasminogen being much stronger. The permeability coefficient (Ks) analyzed with the permeation method revealed that fibrin gels formed in the presence of Lys78-plasminogen were more permeable (porous) than the control gels. The effect on the gel structure was inhibited by the fibrinolytic inhibitor epsilon aminocaproic acid. The same results were obtained in plasma milieu for both mu and Ks as in the purified system, i.e. the gels became more porous with increasing concentrations of Lys78-plasminogen. 3D microscopy pictures of the gels verified the findings. PMID- 10847421 TI - Dose-dependent release of endogenous tissue factor pathway inhibitor by different low molecular weight heparins. AB - Tissue factor pathway inhibitor (TFPI) is released to circulating blood after intravenous and subcutaneous injections of heparins, and may thus contribute to the antithrombotic effect of heparins. A previous study suggested different abilities of various low molecular weight heparins (LMWH) to release endogenous TFPI, but the dose-response relationship was not determined. In the present study, the dose-response relationship for escalating doses of two LMWHs, dalteparin and enoxaparin, on the release of endogenous TFPI was investigated. Six healthy male participants were given 50, 100 and 200 U/kg dalteparin and 0.5, 1.0 and 2.0 mg/kg enoxaparin as a single subcutaneous injection. The study was a randomized, cross-over design with a 1-week wash-out period between each injection. Peak free TFPI antigen and TFPI activity were detected after only 1 h, whereas anti-activated factor X (anti-FXa) and anti-activated factor II (anti FIIa) activities were detected after 2-6 h. Putative therapeutic equivalent doses of dalteparin and enoxaparin gave similar release of endogenous TFPI, but dissimilar effects on anti-FXa and anti-FIIa activities. PMID- 10847422 TI - Influence of exogenous factor VIIa on thrombin generation in cord plasma of full term and pre-term newborns. AB - Factor (F) VIIa has been used to treat adults and children with a variety of bleeding disorders. The results from these studies cannot be extrapolated to newborns because their hemostatic system differs significantly from adults, which may influence the effects of FVIIa on thrombin (IIa) generation. We compared the effects of FVIIa concentrates on IIa generation in plasmas from adults, full-term newborns and pre-term newborns. Defibrinated plasma (using arvin) from adults, or umbilical cords from full-term or pre-term deliveries was supplemented with FVIIa (Novo Nordisk, Bagsvaerd, Denmark), mixed with dilute thromboplastin reagent, and the resultant reaction mixture subsampled periodically into ethylenediamine tetraacetic acid, followed by measurement of total IIa activity (S-2238). Thrombin-alpha2 macroglobulin complexes, determined as residual activity after neutralization with heparin and antithrombin, were subtracted from total IIa to give free IIa. Prothrombin (FII) and inhibitor complexes were measured by enzyme linked immunosorbent assays. Addition of FVIIa caused a reduction in the lag phase for the appearance of free IIa and consumption of FII, which was more pronounced in newborn plasma. There was no increase in peak IIa levels regardless of the amount of FVIIa added. Final inhibitor complex concentrations were increased in plasmas from adults compared with newborns, likely reflecting higher plasma concentrations of FII in adults. Generation of IIa was more rapid in pre term plasma compared with that in adult and full-term cord plasmas due to increased endogenous tissue factor (TF). In summary, FVIIa enhanced IIa generation in plasma from different age groups, with the effect being more pronounced in plasma from pre-term newborns, possibly due to increased levels of plasma TF. PMID- 10847423 TI - The effect of alcohol ingestion on the exercise-induced changes in fibrin and fibrinogen degradation products in man. AB - The present study examined the influence of ingesting a moderate dose of alcohol on plasminogen activator activity (t-PA), plasma fibrinogen (Fb), total degradation products (TDP) and the degradation products of fibrin (FbDP) and fibrinogen (FgDP) at rest and in response to exercise. Eleven male subjects performed two separate experimental trials at an exercise intensity corresponding to 70% maximal oxygen consumption for 35 min. Prior to trials, subjects were either given 0.5 g/kg alcohol in orange-flavoured drink or an equal volume of non caloric non-alcoholic drink 45 min before exercise. Comparison of the levels of t PA, Fb, TDP, FbDP, and FgDP at rest, before and 45 min after the ingestion of alcohol revealed no significant differences between alcohol and control experiments. Exercise resulted in a marked increase in t-PA, TDP, and FgDP, with no appreciable change in FbDP. Although plasma fibrinogen level showed significant decrease post-exercise when subjects ingested alcohol, this difference was small and its biological significance is questionable. While t-PA level increased similarly in response to exercise during alcohol and control trials, a significantly higher response of TDP was found during the control trial compared with alcohol trial. It was concluded that exercise with and without alcohol ingestion is followed by a substantial increase in t-PA, which coincided with an increase in TDP. The increase in TDP was mainly due to an increase in FgDP, but not to FbDP. These findings support the hypothesis that a significant fibrinogenolysis occurs in response to exercise, and moderate intoxication with alcohol prior to exercise reduced this response. PMID- 10847424 TI - Optimization of routine plasma homocysteine monitoring. AB - An elevated plasma homocysteine (Hcy) level is now considered to be an important risk factor in arterial and venous thromboembolic events. As a result of this relatively recent finding, there has been a dramatic increase in the number of requests for Hcy measurement. In our laboratory, this demand has been met by employing an automated immunoassay and improving the pre-analytical handling of blood samples. An automated fluorescent polarization immunoassay (FPIA) gave similar results to a reference high-pressure chromatographic (HPLC) method (r2 = 0.98, enzyme immunoassay = 0.998 HPLC - 0.3) and excellent between-run reproducibility (coefficient of variation <3%). The new assay also required less specialized technical input, and improved the sample throughput two-fold. Pre analytical stability of plasma Hcy concentrations in blood samples is crucial to the accuracy of Hcy monitoring. This stability was improved 10-fold by adopting the anticoagulant acidic citrate instead of ethylenediamine tetraacetic acid for Hcy screening by FPIA. Acidic citrate dramatically inhibits time-related plasma contamination by red-cell Hcy, resulting in improved accuracy and a reduced number of 'spoiled' specimen discards. PMID- 10847425 TI - The use of thrombus precursor protein, D-dimer, prothrombin fragment 1.2, and thrombin antithrombin in the exclusion of proximal deep vein thrombosis and pulmonary embolism. AB - We examined various nonSTAT commercially available coagulation activation markers in an attempt to help diagnose or exclude the often subtle clinical presentations of proximal deep vein thrombosis (PDVT) and pulmonary embolism (PE). Fifty-five patients presenting to the Emergency Department were completely assessed. Eleven patients were diagnosed with PDVT, six patients were diagnosed with PE, and three patients were diagnosed with both PDVT and PE. Thrombus precursor protein (TpP) excluded the diagnosis in 19 of the 35 patients negative for PDVT and/or PE, D Dimer in 15 patients, prothrombin fragment 1.2 in 17 patients, and thrombin antithrombin (TAT) in 14 patients. Both the TpP and TAT enzyme-linked immunosorbent assay (ELISA) tests had 100% sensitivity and negative predictive value for evaluating PDVT and/or PE. The TpP ELISA had the highest specificity (54%) of all four markers studied. PMID- 10847426 TI - Clinical and laboratory expression of associated thrombophilic conditions (homozygous/heterozygous factor V Leiden mutation and heterozygous prothrombin variant 20210A) in an Italian family. AB - Prothrombin variant 20210A is maintained to be a mild risk factor for venous thromboembolism (VTE). The association of this defect with other inherited thrombophilic conditions may result in an increased risk of developing VTE. In this article, a family is described in which prothrombin variant was associated with either homozygous or heterozygous factor V Leiden (FV Leiden) mutation. All family members except the proband were asymptomatic despite the presence and the severity of the underlying genetic defect(s). The proband, who carried homozygous FV Leiden mutation and heterozygous prothrombin variant, experienced recurrent VTE during pregnancies, whereas one brother, with the same defect, was asymptomatic. Mean prothrombin antigen and activity levels were higher in carriers of the prothrombin variant as compared with noncarriers. Thrombin generation was assessed in family members, in carriers of prothrombin variant or homozygous FV Leiden mutation and in a control group. Most of the family members presented with increased prothrombin fragment 1+2 levels possibly because of the presence of the FV Leiden mutation. Although it is conceivable that the co inheritance of prothrombin variant and FV Leiden mutation may increase the risk of VTE, patients with these combined defects may remain asymptomatic. It is likely that acquired triggering conditions play a major role in determining VTE in carriers of a mild genetic predisposition. This has to be taken into account when recommendation for thromboprophylaxis is given. PMID- 10847427 TI - Textilinins from Pseudonaja textilis textilis. Characterization of two plasmin inhibitors that reduce bleeding in an animal model. AB - The incidence of vein-graft occlusion associated with myocardial infarction and thrombosis following the use of the plasmin inhibitor, aprotinin, to reduce blood loss during vascular surgery has prompted the isolation of an alternative kinetically distinct inhibitor of plasmin from the venom of Pseudonaja textilis. This inhibitor has been called textilinin (Txln) and two distinct forms have been isolated from the Brown-snake venom (molecular weight, 6688 and 6692). A comparison of plasmin inhibitor constants for aprotinin and the Txlns 1 and 2 indicated that the former bound very tightly (inhibitor constant, Ki approximately 10(-11) mol/l), while both of the latter bound less tightly (Ki approximately 10(-9) mol/l). Homogeneity of Txlns 1 and 2 was confirmed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and mass spectrometry. A sequence difference of six amino acids was observed between the two forms of Txln. Txln 1 and 2 showed, respectively, 45 and 43% homology with aprotinin, while there was 58 and 55% homology, respectively, with a plasmin inhibitor from the venom of eastern Taipan, Oxyuranus scutellatus. Both Txlns have six cysteines, like other inhibitors of this group, and homology was determined by alignment of these cysteines. Both have been shown to reduce blood loss by about 60% in a murine tail vein bleeding model. It is proposed that the kinetic profiles of Txln 1 and 2 for plasmin allow the arrest of haemorrhage without the possible threat of thrombosis. PMID- 10847428 TI - The effect of plasmapheresis on the serum activity level of dalteparin: a case report. AB - The effect of plasmapheresis on the anticoagulant properties of the low molecular weight heparins has never been studied. We had the opportunity to study this effect in a woman who was receiving dalteparin for the treatment of a pulmonary embolus and required plasmapheresis. Over the course of 8 days, five courses of plasmapheresis were performed. Anti-Xa activity was measured pre- and post dalteparin administration both during and between the first three courses of plasmapheresis. Comparing the rate of change between pairs of anti-Xa activity, the absorption rate was found to have decreased by over 50%. The decay rate, as compared with a historical control, was found to have increased by over 480%. These results suggest that the level of anticoagulation of dalteparin, as monitored through an anti-Xa assay, is reduced by plasmapheresis. PMID- 10847429 TI - East meets West: acute liver failure in the global village. PMID- 10847430 TI - Helicobacter pylori and dyspepsia: still an unresolved controversy? PMID- 10847431 TI - Acute hepatic failure in India: a perspective from the East. AB - Acute hepatic failure (AHF) in India almost always presents with encephalopathy within 4 weeks of the onset of acute hepatitis. Further subclassification of AHF into hyperacute, acute and subacute forms may not be necessary in this geographical area, where the rapidity of onset of encephalopathy does not seem to influence survival. Viral hepatitis is the cause in approximately 95-100% of patients, who therefore constitute a more homogeneous population than AHF patients in the West. In India, hepatitis E (HEV) and hepatitis B (HBV) viruses are the most important causes of AHF; approximately 60% of cases are caused by to these viruses. Hepatitis B virus core mutants are very important agents in cases where hepatitis B results in AHF in this country. Half of the patients with AHF admitted to our centre are female, one-quarter of whom are pregnant. Therefore, pregnant females who contract viral hepatitis constitute a high-risk group for the development of AHF. However, the outcome of AHF in this group is similar to that in non-pregnant women and men. No association with any particular virus has been identified among sporadic cases of AHF. In our centre, approximately one third of AHF patients survive with aggressive conservative therapy, whereas two thirds of deaths occur within 72 h of hospitalization. Cerebral oedema and sepsis are the major fatal complications. Both fungal and gram-negative bacteria are major causes of sepsis. Among patients with AHF, despite the presence of sepsis, its overt clinical features (i.e. fever, leucocytosis) may be absent and objective documentation of the presence of sepsis in such patients is achieved by repeated culture of various body fluids. It should be possible to develop simple, clinical prognostic markers for AHF in this geographical region, in order to identify patients suitable for liver transplantation. PMID- 10847432 TI - Acute hepatic failure: a Western perspective. AB - Acute hepatic failure (AHF) is an uncommon, devastating syndrome, which results in death or the need for liver transplantation in more than 50% of cases. While AHF has numerous causes, most cases are due to viral hepatitis and drug toxicity or idiosyncratic reactions. A significant group with indeterminate causation remains, despite careful investigation. In many of these cases a viral aetiology is suspected, although yet not proven. Major differences exist in the aetiology of AHF between the West and Eastern countries. A wider range of aetiologies exists in the West. Common causes include acetaminophen toxicity and idiosyncratic drug reactions, while viral hepatitis is less frequent. Hepatitis E infection is rarely seen in Western countries in contrast to its high prevalence in the East. The mainstay of AHF management is supportive care in an intensive care unit. Liver transplantation is now the standard of care in many Western liver units for individuals who have a less than 20% probability of survival. Lack of availability of donor livers at short notice remains a significant problem. Methods of liver support used while waiting for a donor liver or for the native liver to regenerate include bioartificial livers, extracorporeal liver assist devices, extracorporeal whole organ perfusion (human and transgenic pig) and hepatocyte transplantation. The effectiveness of these methods remains unproven and awaits controlled clinical trials. Both transplantation and liver support methods require specialized units and expensive and complicated equipment. Further research is necessary to identify modalities of therapy that would be effective as well as widely accessible. PMID- 10847433 TI - Probiotics in infective diarrhoea and inflammatory bowel diseases. AB - Bacteria are present throughout the gastrointestinal tract, but their pattern and concentration vary greatly. Probiotics are living organisms that supply beneficial health effects to the host. So far the beneficial effects of probiotics have been shown, almost exclusively, under poorly defined experimental conditions. There are little convincing data from well-designed, double-blind controlled trials supporting health-promoting effects. The use of probiotics to treat gastrointestinal infections has produced contrasting results. Apart from information on rotavirus infection in children, there is no convincing evidence from controlled studies on the efficacy of probiotics in the prevention or treatment of infective diarrhoea. However, experimental and clinical studies suggest that there are potential therapeutic roles for probiotics in the treatment of inflammatory bowel diseases. This review focuses on the available data concerning the mechanisms of action of probiotics, and on the results from clinical studies using probiotics to treat infective diarrhoea and inflammatory bowel disease. PMID- 10847434 TI - Metronidazole-resistant Helicobacter pylori in an urban Asian population. AB - BACKGROUND: Helicobacter pylori eradication is the mainstay in the treatment of H. pylori-associated peptic ulcer disease. Metronidazole is an important component in most eradication regimens. However, the presence of metronidazole resistant H. pylori adversely affects the efficacy of such regimens. We aimed to study the prevalence of metronidazole resistance in our population, and the factors associated with its presence. METHODS AND RESULTS: From September 1993 to September 1996, 459 H. pylori isolates were collected and analysed. The overall resistance rate was 62.7%. The rate was significantly higher among women compared with men (P < 0.05). When the results were analysed according to each year, there was a significant increase in the resistance rate from 50.5% in the first year to 72.7% in the third year (P = 0.0039). CONCLUSIONS: There was a significant rise in the prevalence of metronidazole resistance over a 3-year period. The presence of metronidazole resistance adversely affects the eradication rates of nitroimidazole-based regimens. Hence, in a population with a high prevalence of metronidazole resistance, the use of non-nitroimidazole-based therapy may be more efficacious. This information on resistance rates is important for the empirical choice of antibiotic against H. pylori in a population. PMID- 10847435 TI - Clinical presentation of Helicobacter pylori-positive and -negative functional dyspepsia. AB - BACKGROUND: A questionnaire was used to record the clinical presentation of functional dyspepsia in relation to Helicobacter pylori infection in a consecutive series of patients sent for upper gastrointestinal endoscopy. Only patients without macroscopic abnormalities in their oesophagus, stomach and duodenum were included. METHODS: The study questionnaire included two questions related to daily life, and the calculation of a symptom score. Biopsy specimens were taken from all patients for histological and microbiological examination, and immunoglobulin G antibodies were also determined. RESULTS: Two hundred and twenty-two patients were H. pylori positive and 182 patients were H. pylori negative. Loss of weight was significantly more common in the H. pylori positive group (P<0.001). Patients with H. pylori infection had a significantly higher overall symptom score compared with H. pylori-negative subjects (P<0.05). In addition, the severity of epigastric and nocturnal pain, heartburn, retrosternal heartburn, and vomiting was significantly higher in H. pylori-positive functional dyspeptic patients, and the influence on daily life and activities was significantly worse. CONCLUSIONS: The combination of retrosternal pain, weight loss, food intolerance and the absence of halitosis signified a 64% accuracy in predicting H. pylori infection. It is not possible to differentiate between H. pylori-positive and H. pylori-negative functional dyspeptics on the basis of clinical presentation and the number of complaints. However, overall symptom score and severity of several symptoms was significantly higher in the H. pylori positive group. PMID- 10847436 TI - Gastroallergic anisakiasis: findings in 22 patients. AB - BACKGROUND AND AIMS: Ingestion of Anisakidae larvae in raw seafood may cause anisakiasis. However, despite the high level of consumption of seafood in Spain, only a few cases of anisakiasis have been reported until now. Anisakis simplex can cause allergic reactions in sensitized patients as a result of its parasitism in the gastrointestinal tract. The purpose of this study was to analyse the clinical findings in 22 patients with gastroallergic anisakiasis. METHODS: Patients with allergic and/or gastric symptoms after seafood ingestion were evaluated in the emergency room of the La Paz General University Hospital. Skin testing for Anisakis simplex and tests on the implicated seafood were performed and amounts of serum-specific immunoglobulin E were assessed. A gastroscopy was performed in those patients with severe allergic or/and persistent gastric symptoms after ingestion of raw or undercooked seafood. RESULTS: Twenty-two patients were diagnosed with gastroallergic anisakiasis in 1 year. Most patients presented to the emergency room of our hospital with allergic symptoms. Gastric symptoms were usually moderate. Gastroscopy revealed local mucosal oedema and gastric erosion at the point of fixation. Two or more worms were detected in three patients. The mean time of latency of allergic symptoms was 5 h, while the mean time for gastric symptoms was 3 h. CONCLUSION: Anisakis simplex parasitism was the causative agent of allergic and gastric symptoms. Gastroallergic anisakiasis appears to be a relatively common disease, that may have been underdiagnosed. PMID- 10847437 TI - Surgery for gastric carcinoma in patients more than 85 years of age. AB - BACKGROUND: The incidence of gastric carcinoma is increasing in elderly patients. It has not been determined whether surgery improves the quality of life or prolongs survival in patients older than 85 years. This study was designed to evaluate surgery as a treatment option in patients more than 85 years of age. METHODS: The records of 18 patients aged 85 to 91 years (mean 87.3 years), who underwent surgery for gastric carcinoma between 1983 and 1997 were analysed. RESULTS: Three patients had multiple lesions. A total of 21 lesions were examined. Operative procedures included distal gastrectomy (n = 12), total gastrectomy (n = 4), proximal gastrectomy (n = 1), and laparoscopic wedge resection (n = 1). Perigastric lymphadenectomy was performed in 15 patients. No lymph node dissection was performed in three patients. Postoperative complications, including delirium, respiratory dysfunction, cardiac dysfunction, anastomotic leakage, bleeding, and ileus, occurred in 11 patients. There were two hospital deaths in patients who underwent emergency surgery. Survival was neither shortened nor prolonged by surgery as determined by life table analysis. CONCLUSIONS: Surgical treatment should not be avoided based solely on the age of the patient, and quality of life in this population may be improved by surgery. Careful patient selection should include an assessment of the will to live. PMID- 10847438 TI - Decreased trabecular bone mineral density in newly diagnosed inflammatory bowel disease patients in Korea. AB - BACKGROUND: Decreased bone mineral density (BMD) is common in Western patients with inflammatory bowel disease (IBD). However, BMD has never been studied in Asia where the demographic and socio-economic status are different from the West. The aim of this study was to investigate the prevalence and mechanisms of osteopenia in newly diagnosed Korean patients with IBD. METHODS: We studied 14 patients with Crohn's disease (CD) and 25 patients with ulcerative colitis (UC), all of whom had never been treated with corticosteroids. Bone mineral density was measured in the lumbar spine and the femoral neck by dual energy X-ray absorptiometry. Biochemical parameters including serum osteocalcin, parathyroid hormone, plasma inactive and active vitamin D, and urinary deoxypyridinoline were measured. RESULTS: The BMD Z score at the lumbar spine was lower both in CD and in UC patients, but there was no significant difference between the two groups. There was no significant difference in nutritional status or biochemical parameters of bone metabolism between patients with a normal BMD and those with a decreased BMD. CONCLUSIONS: Low BMD at the lumbar spine is common in newly diagnosed Korean patients with IBD, a result which is similar to Western studies. The mechanism for low bone mass remains undetermined; however, nutritional status and hormonal parameters of bone metabolism, and ethnic differences are not likely to be an important factor in the pathogenesis of this bone loss. PMID- 10847439 TI - Identification of a pre-S2 mutant in hepatocytes expressing a novel marginal pattern of surface antigen in advanced diseases of chronic hepatitis B virus infection. AB - BACKGROUND AND AIMS: The expression of hepatitis B viral (HBV) antigens in liver tissue reflects the replicative status of chronic HBV infection. We have previously recognized a novel marginal pattern of hepatitis B surface antigen (HBsAg) in hepatocytes, which usually clusters in groups and emerges at the late non-replicative phase. This study was designed to investigate whether the marginal-type HBsAg represented the gene product of a specific HBV-surface mutant. METHODS: Microdissection of cirrhotic nodules homogeneously expressing marginal HBsAg was performed on two of 12 resected livers from HBsAg-seropositive patients with hepatocellular carcinoma. The gene presumably encoding marginal HBsAg was polymerase chain reaction (PCR)-cloned, sequenced and analysed. In vitro transfection and expression of the cloned surface mutant plasmids were performed on the Huh7 cell line to illustrate intrahepatic HBsAg expression. RESULTS: Immunohistochemical staining revealed that the marginal HBsAg was positive for pre-S1 and thus contained large surface proteins. The PCR cloning and sequencing of the genes presumably encoding marginal-type HBsAg in both cases revealed the same deletion at the 5' terminus (nt 2-55) of pre-S2. A point mutation on the small-surface (S) antigen was also found in one case. The pre-S2 deletion sequence and the mutation sites of the S gene coincide with human lymphocyte antigen-restricted T- and/or B-cell epitopes. In vitro transfection of the mutant plasmid revealed a blot-like retention or accumulation of HBsAg in the cytoplasm or at the periphery of hepatocytes, accompanied by a decreased secretion of HBsAg in the culture supernatant, mimicking intrahepatic expression. CONCLUSION: A natural pre-S2 deletion mutant was identified in hepatocytes expressing a novel marginal pattern of HBsAg, which probably contains mutant, large, surface proteins. The biological significance of the pre-S2 deletion mutant should be interesting in view of the clustering proliferation of hepatocytes expressing marginal HBsAg. PMID- 10847440 TI - Enhancement of poly-adenosine diphosphate-ribosylation in human hepatocellular carcinoma. AB - BACKGROUND: Poly-adenosine diphosphate (ADP)-ribosylation, catalysed by poly(ADP ribose) polymerase (PARP), is a post-translational modification of nuclear proteins and is involved in a wide range of biological processes including DNA repair, cell proliferation and malignant transformation. Alteration of this reaction in human hepatocellular carcinoma (HCC) is of interest, but has not yet been explored. The aim of this study was to evaluate poly-ADP-ribosylation and to compare the expression of PARP in HCC and adjacent non-tumour tissues. METHODS: Tumorous and adjacent non-tumorous tissues were obtained from five consecutive patients with HCC during surgery for tumour resection. Tissue homogenates were subjected to ADP-ribosylation with [32P]-nicotinamide adenine dinucleotide. The ADP-ribosylated proteins were separated by sodium dodecylsulfate-polyacrylamide gel electrophoresis, followed by autoradiography. Expression of PARP was also evaluated by western blotting. RESULTS: Several proteins were ADP-ribosylated in human HCC tissues. Notably, the radiolabelling of a 116-kDa protein was remarkably greater than that in adjacent non-tumorous tissues (86.5 +/- 35.2 arbitrary units by densitometry vs 12.2 +/- 9.9, mean +/- SD, n = 5, P < 0.02). The radiolabelling of the 116-kDa protein was decreased in the presence of PARP inhibitors in a concentration-dependent manner. Immunoblot analyses revealed that the radiolabelled protein was PARP and that its expression was significantly greater in HCC than in adjacent non-tumorous tissues (333 +/- 204% of non tumorous tissue, P < 0.05). CONCLUSIONS: We found that poly-ADP-ribosylation and PARP expression were significantly increased in human HCC compared with those in adjacent non-tumorous tissues in surgically obtained specimens. PMID- 10847441 TI - Gastrointestinal metastasis in hepatocellular carcinoma: radiological and endoscopic studies of 11 cases. AB - BACKGROUND: From October 1990 to January 1999, 11 of 2237 hepatoma patients at our hospital had gastrointestinal tract metastasis. We describe the radiological and endoscopic features, clinical course and prognoses of those patients. METHODS: All patients were male. Six patients were hepatitis B carriers, and cirrhosis was noted in eight patients on admission. We reviewed all available radiological, endoscopic and pathological features. RESULTS: The commonest clinical presentation was frank gastrointestinal bleeding. Histological proof of gastrointestinal involvement was seen in six patients. Endoscopic features included ulcerative tumours mimicking advanced gastric carcinoma (43%) and submucosal tumours (29%). The sites of organ involvement were stomach (five), duodenum (two), colon (three) and duodenum and colon (one). Direct invasion by a contiguous neoplasm was the major route of gastrointestinal tract metastasis. Portal vein thrombosis may be the key point of haematogenous spread to other sites. CONCLUSIONS: The prognosis in these patients was extremely poor. Almost all patients died within 5 months if no further aggressive management was performed. Surgical intervention may be the optimal choice for palliative treatment of HCC with gastrointestinal tract involvement. PMID- 10847443 TI - Budd-Chiari syndrome: our experience of 71 patients. AB - BACKGROUND: Hepatic venous outflow obstruction (Budd-Chiari syndrome) is frequently encountered as a cause of portal hypertension at our centre. METHODS AND RESULTS: We studied the clinical presentation, therapeutic modalities and outcome of 71 patients with hepatic venous outflow obstruction between 1992 and 1997. Twenty-seven patients presented with acute disease, while 44 had chronic presentation. Abdominal pain, distension, jaundice and upper gastrointestinal bleeding were the commonest presenting symptoms. The majority of patients had distended veins, hepatomegaly, splenomegaly, ascites and ankle oedema. The diagnosis was made on the basis of inferior vena cavography/functional hepatography and pulsed Doppler ultrasonography and/or liver biopsy in 39 patients and pulsed Doppler ultrasonography and/or liver biopsy in 32 patients. Pulsed Doppler ultrasonography accurately detected the site of the block in 31 of 39 patients (79.4%). The obstruction was in the hepatic vein in 20 patients, in the inferior vena cava in 10, and in both in 41 patients. Aetiologically, four had pregnancy-related disease, four tumour-related, three hypercoagulable states, 18 inferior vena cava membranes and 42 were idiopathic. Of 30 patients in whom liver biopsy was carried out, eight had centrizonal congestion and necrosis, 13 had mixed features and nine had established cirrhosis. Seven patients underwent a shunt operation and surgical membranotomy was carried out in one. Three patients (4.2%) died in the hospital. CONCLUSIONS: Hepatic venous outflow obstruction is a common problem; patients present with abdominal pain, distension, jaundice, distended veins, ascites and ankle oedema. Chronic presentation is more frequent. Pulsed Doppler ultrasound, venography and liver biopsy are very helpful in diagnosis. PMID- 10847442 TI - Liposome-encapsulated OK-432 specifically and sustainedly induces hepatic natural killer cells and intermediate T cell receptor cells. AB - BACKGROUND: OK-432 is a biological response modifier used in Japan to augment host immunity and is known to increase the host antitumour response. By using liposomes, which are vesicles made from phospholipids that have a structure resembling the cell membrane, we encapsulated OK-432. METHODS AND RESULTS: Encapsulated OK-432 was injected into the tail veins of mice, and its effect was compared with that of unencapsulated OK-432 given intravenously. In mice that received either form of OK-432, both the number of natural killer (NK) and intermediate T cell receptor (intTCR) cells (intrahepatic T cells generated by extrathymic differentiation) increased markedly in the liver, with the peak level occurring 3 days after administration. Both forms of OK-432 also increased cytotoxic activity against Yac-1 cells. The increase in numbers of cells and in cytotoxic activity in the liver persisted for longer in mice that received encapsulated OK-432 than in animals that received unencapsulated OK-432. CONCLUSIONS: Because it has been shown that both NK and intTCR cells play an important role in tumour immunity, an increase in the number of such cells can be considered likely to have an increased antitumour effect. Encapsulated OK-432 elicited liver-specific augmentation of cytotoxic activity and the effect was more persistent than that produced by OK-432 given in the conventional form; therefore, it may be useful for the treatment of tumours, particularly those arising in the liver. PMID- 10847444 TI - School absenteeism after upper gastrointestinal endoscopy in children. AB - BACKGROUND AND AIMS: To examine the functional impact of upper gastrointestinal endoscopy as a day procedure, particularly in relation to subsequent school attendance. METHODS: Symptoms and morbidities were prospectively recorded from school-aged children during observation in hospital and for 3 days at home after endoscopy by using a structured questionnaire. Reasons for school absence were identified. RESULTS: Sixty children (31 boys, 29 girls) were enrolled in the study (mean age 10.6 +/- 2.8 years, range 6.1-16.2 years). Following the procedure, symptoms were reported at home in 68.3% (same day), 56.7% (day 1) and 20% (day 2).The commonest symptoms were sore throat, tiredness and dizziness. Twenty-nine children (48.3%) did not attend school on the day following the procedure but most (26 of 29) had returned to school by the second day. The main reason for their absence was residual physical discomfort related to the procedure (55.2%). CONCLUSIONS: Persisting physical discomfort and school absenteeism are common following upper gastrointestinal endoscopy in children. PMID- 10847445 TI - Images of interest. Hepatobiliary and pancreatic: computed tomography in liver disease. PMID- 10847446 TI - Images of interest. Gastrointestinal: what's in a hernia? PMID- 10847447 TI - Sustained response to combination therapy in a patient with chronic hepatitis C and thrombocytopenia secondary to alpha-interferon. AB - Recent data suggest that hepatitis C viral (HCV) infection may induce a significant autoimmune reaction to platelets, but the mechanism is unknown. Many patients with chronic hepatitis C, in fact, have high levels of platelet associated immunoglobulin G (PAIgG) and HCV-RNA is present in the platelets of 100% of those patients with thrombocytopenia and high PAIgG levels. Hepatitis C virus infection has been associated with the development of thrombocytopenic purpura, sometimes triggered during interferon (IFN) therapy. In such cases, the treatment of the underlying disease is a difficult problem to solve. We report the case of a patient with chronic hepatitis C, who developed life-threatening thrombocytopenic purpura after a prolonged course of IFN-alpha2b over a 4-year period. Treatment with anti-immunoglobulin gammaglobulin (Polyglobin; Quimica Farmaceutica Bayer, Barcelona, Spain) had a transient effect on the platelet count, but prolonged therapy with prednisone was necessary for definitive relief of the haematological complication. Two years later, the patient was treated with combined therapy, including ribavirin (1200 mg/day) and IFN-alpha2b (5 mU, t.i.w.) for 12 months. This therapy induced a sustained response, both biochemical and virological, without haematological complications. This observation suggests that ribavirin may be of benefit in the treatment of immune mediated thrombocytopenia in patients with chronic hepatitis C, preventing the harmful effect of IFN-alpha but also allowing both drugs to be combined so as to increase the probability of sustained remission of the liver disease. PMID- 10847448 TI - Wilson's disease with superimposed autoimmune features: report of two cases and review. AB - We describe two females, 15 and 23 years old, respectively, who presented with classical features of Wilson's disease (WD) and several features of autoimmune hepatitis (AIH). The first patient was initially diagnosed as AIH and treated with prednisolone which caused clinical improvement, with an increase of serum albumin from 22 to 30 g/L, and a decrease of aspartate aminotransferase from 103 to 47 U/L. Subsequent diagnosis of WD and introduction of penicillamine gave excellent improvement and complete normalization of liver function tests. The second patient, at first also diagnosed as having AIH, was treated with steroids and azathioprine with initial improvement, but subsequent deterioration. The diagnosis of WD was made 2 years after initial diagnosis of AIH, as the patient reached end-stage liver disease and required a transplant. Therefore, d penicillamine treatment was not attempted. We conclude that, in patients with AIH, a thorough screening for WD is necessary, particularly when the response to steroid therapy is poor. Conversely, in patients suffering from WD with superimposed features of AIH, a combination of steroids and penicillamine may be of benefit. PMID- 10847449 TI - National plan against cancer in France. PMID- 10847450 TI - "Allegre, resign!" demand demonstrators. PMID- 10847451 TI - Lancet against overoptimism about cancer. PMID- 10847452 TI - Does the French government forget public hospitals? PMID- 10847453 TI - Republican presidential candidates capitalize on alternative medicine. PMID- 10847454 TI - Artificial neural networks: a new model for assessing prognostic factors. PMID- 10847455 TI - Downstream molecular determinants of response to 5-fluorouracil and antifolate thymidylate synthase inhibitors. AB - Thymidylate synthase (TS) is an essential enzyme for the de novo synthesis of thymidylate and subsequently DNA synthesis. TS has been used as a target for cancer chemotherapy in the development of fluoropyrimidines such as 5 fluorouracil (5-FU) and 5-fluorodeoxyuridine and of novel folate-based TS inhibitors such as ZD1694 (Tomudex, Raltitrexed), ZD9331, LY231514 (ALIMTA, Pemetrexed), AG337 (Thymitaq, Nolatrexed) and AG331. Although TS has been considered as a target for chemotherapy, the precise mechanism by which TS inhibition leads to cell death is still not completely resolved. TS inhibition results in depletion of dTTP, an essential precursor for DNA, and an increase in dUTP. This results in the so-called thymine-less death due to misincorporation of dUTP into DNA; its excision, catalysed by uracil-DNA glycosylase, results in DNA damage. Both this imbalance in dTTP/dUTP and DNA damage can result in induction of downstream events, leading to apoptosis. On the other hand a specific interaction exists between oncogenes and TS, by binding of TS protein to the p53 and c-myc RNA, while wt p53 can also inhibit TS promotor activity. TS inhibition by either 5-FU or antifolates can also result in a depression of TS protein mediated inhibition of TS mRNA translation leading to induction of more TS protein synthesis, and p53 protein may further deregulate this process. These complex indirect and direct interactions between oncogenes and TS may have as yet unclear clinical implications, since most data are based on in vitro or in vivo studies and some results are contradictive. In some preliminary clinical studies evidence was postulated for a combined prognostic role for TS and p53. This knowledge should be used to design clinical studies with the aim to deliver effective treatment to potentially sensitive patients both in the adjuvant setting and in advanced stage disease. PMID- 10847456 TI - P53 determination alongside classical prognostic factors in node-negative breast cancer: an evaluation at more than 10-year follow-up. AB - BACKGROUND: There is heterogeneity of methods and conflicting results concerning the prognostic value of p53 in node-negative breast cancer. The clinical value of a quantitative method for measuring tumoralp53 content still needs to be evaluated. PATIENTS AND METHODS: A long-term retrospective study was conducted on 297 node-negative patients with a median follow-up greater than 10 years (11 years, 101-172 months). Classic prognostic factors were considered including age, tumor size, histoprognostic grade and estradiol (ER) and progesterone receptors (PR). In addition, the value of p53 determination (immunoluminometric assay in tumor cytosol) was assessed for this long follow-up period. RESULTS: p53 concentrations were significantly linked to the histological grade (P = 0.001), to tumor size (P = 0.02) and ER status (P = 0.01). Higher p53 tumoral concentrations were found in tumors with large size, pejorative histological grade and negative ER status. In contrast, p53 tumoral concentrations were not influenced by menopausal or PR status. Multivariate Cox analysis demonstrates that tumor size was the only significant predictor of disease-free survival (P = 0.049) with a risk factor at 1.38. As regards specific survival, univariate Cox analysis indicates that p53 taken as a continuous variable is a significant predictor (P = 0.024) together with histological grade, tumor size and ER status. In a multivariate Cox analysis there were two significant and independent variables for predicting overall survival: tumor size (P = 0.031) and, ER status (P = 0.015) with the highest risk factor (RR = 2.14). CONCLUSIONS: The present investigation points out that the prognostic power of p53 tumor determination evaluated at more than 10 years median survival is not higher than the well recognized classic prognostic factors in node-negative breast cancer. The present data highlight the need to assess the prognostic value of potentially new biological factors in node-negative breast cancer on cohorts of patients followed over periods in excess of 10 years. PMID- 10847457 TI - Response criteria for NHL: importance of 'normal' lymph node size and correlations with response rates. AB - BACKGROUND: Oncologic literature cites many different definitions of critical response measurements. PATIENTS AND METHODS: Response criteria (RC) for non Hodgkin's lymphoma (NHL) were developed by lymphoma experts, endorsed by international lymphoma clinicians, and applied to a 166-patient rituximab (Rituxan, MabThera) trial by a third-party, blinded panel of NHL experts (LEXCOR). Retrospectively, we analyzed this data using variations of the original RC and comparing with recently published RC. RESULTS: The definition of a 'normal' lymph node affected the complete response (CR) rate (< or = 1.0 x 1.0 cm, 6%; < or = 1.5 x 1.5 cm, 18%; < or = 2.0 x 2.0 cm, 28%); overall response rate (ORR) was not affected. CR rates increased progressively without > or = 28 days response confirmation: 12% vs. 6% (< or = 1.0 x 1.0 cm), 26% vs. 18% (< or = 1.5 x 1.5 cm), and 36% vs. 28% (< or = 2.0 x 2.0 cm). CR rate and duration of response (DR) were unaffected when only the six largest, rather than all lesions, were measured. When the new RC were applied, CR rate (32%) was higher and DR (13.9 months) and time to progression (15.6 months) were shorter in complete responders. CONCLUSIONS: Standard RC must be consistently and rigorously applied for accurate comparisons between studies. PMID- 10847458 TI - A phase II study of paclitaxel in chemonaive patients with recurrent high-grade glioma. AB - BACKGROUND: The prognosis of malignant gliomas remains poor. In recurrent disease, chemotherapy can be considered. PATIENTS AND METHODS: In this phase II study we determined the anti-tumour efficacy of paclitaxel 200 mg/m2 in a three hour intravenous infusion every three weeks in chemonaive patients with recurrent high-grade glioma in terms of response, survival, and quality of life. RESULTS: In 17 patients (14 glioblastoma multiforme, 3 anaplastic astrocytoma) 69 paclitaxel cycles were administered. Partial or complete responses were not observed. Stable disease for four to six months was observed in five patients (29%). Median time to progression and median survival were two and 10 months, respectively. Toxicity due to paclitaxel was as to be expected and minor in most cases. Quality of life and mood estimates appeared rather stable over time. CONCLUSIONS: We conclude that three-weekly 200 mg/m2 paclitaxel chemotherapy for patients with recurrent high-grade gliomas did not lead to major complications or adverse effects on quality of life and mood. However, this therapy is of only very limited value in terms of response and survival in such patients. PMID- 10847459 TI - A multicenter phase II study of a five-day regimen of oral 5-fluorouracil plus eniluracil with or without leucovorin in patients with metastatic colorectal cancer. AB - PURPOSE: To evaluate the safety and efficacy of a five-day regimen of oral 5 fluorouracil (5-FU) plus eniluracil (776C85) in patients with metastatic colorectal cancer (CRC). PATIENTS AND METHODS: Seventy-five patients with metastatic CRC that was previously untreated or refractory to 5-FU-leucovorin (LV) were enrolled and divided into two strata based upon their treatment history. Twenty-four had not previously received chemotherapy or had received adjuvant chemotherapy that ended > 6 months prior to enrollment on study (previously untreated stratum). Fifty-one patients had disease refractory to intravenous (i.v.) 5-FU-LV (previously treated stratum). All patients received seven consecutive daily doses of eniluracil (20 mg/day) with once daily oral 5-FU given on days 2-6, repeated every four weeks. One-half of the patients in each stratum also received 50 mg/day oral LV on days 2-6. The 5-FU dose was 25 mg/m2 when administered without LV and 20 mg/m2 when administered with LV. RESULTS: Partial response (PR) was noted in 2 of 12 patients receiving eniluracil-5-FU and in 3 of 12 patients receiving eniluracil-5-FU-LV in the previously untreated stratum. No responses were observed in the refractory disease stratum, however, 15 patients (30%) demonstrated stable disease over 2-18+ courses of therapy. Non hematologic toxicities were mild; only 7% of patients experienced grade 3 diarrhea. Myelosuppression was frequent and dose limiting. Neutropenic sepsis was reported in 13.5% of patients. CONCLUSIONS: Eniluracil with 5-FU administered orally with or without LV on a five-day schedule is active and well tolerated when given as primary therapy to patients with metastatic CRC. PMID- 10847460 TI - A phase I trial of docetaxel and gemcitabine in patients with advanced cancer. AB - BACKGROUND: Docetaxel and gemcitabine are active in a broad range of malignancies. The objective of this phase I trial was to determine the maximally tolerated doses of the combination of docetaxel and gemcitabine. PATIENTS AND METHODS: Patients with advanced cancer, WHO performance status 0-2, who had received up to one prior chemotherapy regimen were treated with gemcitabine on days 1 and 8 and docetaxel on day 8 repeated every 21 days. Prophylactic ciprofloxacin was commenced on day 11 of each cycle and continued until the neutrophil count reached 1.0 x 10(9)/l. G-CSF was not administered. Dose levels studied were docetaxel/gemcitabine: 60/800, 60/1000, 75/1000, 75/1200, 85/1200 and 100/1200 mg/m2. RESULTS: Thirty-nine patients were entered and all were assessable for toxicity. The highest administered dose level was 100 mg/m2 docetaxel and 1200 mg/m2 gemcitabine with dose limiting toxicities of febrile neutropenia, grade 4 neutropenia > or = 7 days, grade 4 thrombocytopenia, grade 3 stomatitis and/or grade 3 fatigue in three out of six patients. Treatment was well tolerated (40 cycles) in the 10 patients treated at the recommended dose level (85/1200) with only a single episode of febrile neutropenia and grade 3 or 4 non-hematologic toxicity was infrequent. There was no significant pulmonary toxicity. Responses were seen in a range of malignancies including non-small-cell lung cancer. CONCLUSIONS: The recommended dose level of 85 mg/m2 docetaxel and 1200 mg/m2 gemcitabine has a favourable toxicity profile and is suitable for further investigation in phase II trials. This non-platinum containing regimen warrants further investigation as a potential alternative to platinum containing regimens in non-small-cell lung cancer and other malignancies. PMID- 10847461 TI - High-dose chemotherapy and autologous haematopoietic support in poor risk non seminomatous germ-cell tumours: an effective first-line therapy with minimal toxicity. AB - BACKGROUND: The prognosis of patients with high-risk germ-cell cancer is poor. The toxicity and efficacy of first-line high-dose chemotherapy (HDCT) with stem cell support was evaluated, following induction chemotherapy with BEP. PATIENTS AND METHODS: Twenty patients with poor prognosis non seminomatous germ-cell tumour by the International Consensus prognostic criteria received induction with BEP followed by one cycle of HDCT (CEC) given with carboplatin (1800 mg/m2), etoposide (1800 mg/m2), and cyclophosphamide (140 mg/kg). Of the above 20 patients only 3 received a second cycle of HDCT. Peripheral blood stem cells were infused on day 0. RESULTS: Twenty patients were assessable for toxicity and response. After a median follow-up of 27 months 15 patients (75%) are alive, 12 (60%) are disease free and 3 (15%) are alive with disease. Median survival has not been reached and overall survival at four years is 66% with a durable complete response rate of 50%. There were no deaths or cases of severe toxicity. Median time to a granulocyte count > 500/microl and platelets > 20,000/microl was 10 and 12 days respectively. Five patients have died from progressive disease 5 35 months after HDCT. CONCLUSIONS: These results support the case of first-line HDCT. The excellent toxicity profile of BEP/CEC and the two-year overall survival of 78% are encouraging and support further the ongoing randomised US intergroup study evaluating high-dose CEC after BEP. PMID- 10847462 TI - Front-line treatment of advanced non-small-cell lung cancer with MTA (LY231514, pemetrexed disodium, ALIMTA) and cisplatin: a multicenter phase II trial. AB - BACKGROUND: To evaluate the activity of MTA plus cisplatin in chemotherapy-naive patients with non-small cell lung cancer (NSCLC). PATIENTS AND METHODS: Thirty six chemotherapy-naive patients with NSCLC received 500 mg/m2 MTA plus 75 mg/m2 cisplatin every 21 days, with 4 mg dexamethasone orally twice daily on the day before, of, and after MTA administration. RESULTS: Median age was 58 years. WHO performance status was 0-2. Eighteen patients each had stage IIIB and IV disease. Seventeen patients each had squamous-cell and adenocarcinoma; two had undifferentiated disease. Fourteen patients (39%; 95% confidence interval: 23% 57%) showed partial response; seventeen (47%) had stable disease. Median survival was 10.9 months. Twenty-one patients (59%) experienced grade 3 or 4 granulocytopenia without fever or infection. Five (14%) and six (17%) patients experienced grade 3 anemia and grade 3 or 4 thrombocytopenia, respectively. Nonhematological toxicities included grade 3 nausea in two patients (6%), and grade 3 and 4 diarrhea in one patient (3%) each. One patient each experienced grade 4 ALT and grade 3 bilirubin and AST elevations. CONCLUSIONS: MTA plus cisplatin is well tolerated and active against NSCLC. Further studies of this combination are warranted. PMID- 10847463 TI - Clinical, hematological and cytogenetic characteristics of atypical chronic myeloid leukemia. AB - BACKGROUND: Atypical chronic myeloid leukemia (aCML) is an infrequent chronic myeloproliferative disorder characterized by leukocytosis, absence of Philadelphia chromosome or BCR-ABL rearrangement, and marked myeloid dysplasia. Some cases have an absolute monocytosis but can be distinguished from chronic myelomonocytic leukemia (CMML) by the presence of a higher percentage (> 15%) of circulating immature granulocytes. PATIENTS AND METHODS: In a series of 11 patients with a diagnosis of aCML according to the FAB proposals we have analyzed the most relevant clinical, hematological and cytogenetic characteristics. RESULTS: The median age was 65 years (16-84). All but one case showed, at time of diagnosis, leukocytosis (median WBC was 36 x 10(9)/l), 55% had moderate anemia and 36% had thrombocytopenia. Most cases had marked dysplasia, particularly in the granulocytic lineage (82% of the cases), and all cases showed bone marrow red hypoplasia. Cytogenetic abnormalities were present in 9 out of the 11 patients. Trisomy 8 was observed in three cases and other clonal chromosomal abnormalities included deletions of 5q, 13q, 17p, 12q, and 11q as well as a t(6;8)(p23;q22) translocation. Fluorescence in situ hybridization (FISH) studies failed to demonstrate ETV-6 gene involvement. The median survival time from diagnosis was only 14 months (range 3-56 months). CONCLUSIONS: These data suggest that aCML is a rare disease which is characterized by leukocytosis, with dysgranulopoiesis, BM erythroid hypoplasia, chromosomal, though not recurrent, abnormalities and poor prognosis. PMID- 10847464 TI - Phase II study of concurrent chemotherapy and radiotherapy for unresectable stage III non-small-cell lung cancer: long-term follow-up results. Japan Clinical Oncology Group Protocol 8902. AB - BACKGROUND: Although chemoradiotherapy is standard treatment for unresectable stage III non-small-cell lung cancer (NSCLC), few long-term survival data exist. PATIENTS AND METHODS: Between October 1989 and December 1991, 74 patients with histologically or cytologically proven NSCLC, unresectable stage IIIA or IIIB, were entered into this study. Seventy patients were eligible and evaluable for response, toxicity, and survival analysis. Chemotherapy consisted of cisplatin (100 mg/m2 on days 1, 29, and 57) and vindesine (3 mg/m2 on days 1, 8, 29, 36, 57, and 64). Thoracic radiotherapy was administered for two weeks (2 Gy given 10 times, five fractions per week), and after a 14-day rest period, the previous schedule of radiotherapy was repeated for two weeks. A 10-Gy to 20-Gy dose of radiotherapy was administered during the third cycle of chemotherapy. RESULTS: Of the 70 evaluable patients, 1 (1.4%) had a complete response (CR) and 51 (72.9%) had a partial response (PR). The median survival time was 14.8 months, and the five-year survival rate was 14.8%. The major toxicity was leukopenia (> or = grade 3, 93%). Other toxicities > or = grade 3 included anemia (34%), nausea/vomiting (27%), alopecia (7%), thrombocytopenia (4%), and serum creatinine elevation (1%). Treatment related death occurred in two patients (2.8%). One patient died of pneumonia and pneumothorax, and the other of hemoptysis. CONCLUSIONS: Concurrent chemotherapy and radiotherapy has the potential to provide long-term survival with acceptable toxicities. PMID- 10847465 TI - A comparison of methods currently used in clinical practice to estimate familial breast cancer risks. AB - BACKGROUND: With the identification of genes predisposing to hereditary breast cancer, the accurate and consistent estimation of a woman's risk of developing breast cancer based on her family history is of paramount importance if national service guidelines are to be developed. PATIENTS AND METHODS: The residual lifetime risk of developing breast cancer was estimated for 200 women attending a breast cancer genetic assessment clinic by three different methods currently in use in the UK. Risks were computed on the basis of the Cancer and Steroid Hormone (CASH) study data and were classified as 'low/moderate' (<20%) or 'high' (>20%). These risk categories are representative of those currently used to allocate surveillance and genetic testing. Risks were then compared to estimates derived by other methods used in current clinical practice, including those of Houlston and Murday. RESULTS: The CASH data-based method ascribed 27% to the high risk category, as compared to 53% for the combined Houlston and Murday methods. A method based on the number of affected relatives alone ascribed only 14% to the high risk category. Overall, 108 (54%) women were placed in the same risk category by all three methods. CONCLUSIONS: This study demonstrates that there is a significant degree of variability between methods currently used to estimate breast cancer risk which has serious implications for individual patient management, service provision and multicentre studies evaluating the benefits of genetic testing for breast cancer susceptibility. PMID- 10847466 TI - Phase I study with weekly cisplatin-paclitaxel and concurrent radiotherapy in patients with carcinoma of the cervix uteri. AB - BACKGROUND: Cisplatin and paclitaxel are active in cervical cancer and both are able to potentiate the effects of radiotherapy. In this study we evaluated the maximum-tolerated dose (MTD) of paclitaxel in combination with a fixed dose of cisplatin when given weekly concurrently with pelvic radiotherapy to patients with carcinoma of the cervix uteri. PATIENTS AND METHODS: Eighteen patients with cervical cancer were enrolled in this study. Cisplatin (30 mg/m2) and paclitaxel (starting dose 40 mg/m2; 5 mg/m2 escalation per level) were given on day 1 of radiotherapy and then weekly for six times. Radiotherapy was given to the pelvis with a four-field box technique for five days each week. Patients received 65 Gy in 1.8 Gy fractions. Cohorts of three patients were enrolled at each level and three further patients were included if one or two dose-limiting severe adverse events (SAE) were recorded. SAE was defined as grade 3 or 4 nonhematologic toxicity, excluding nausea or vomiting and alopecia, grade 4 neutropenia or thrombocytopenia, and prolonged (> 1 week) neutropenia or thrombocytopenia. RESULTS: Four levels were studied (paclitaxel 40, 45, 50, 55 mg/m2) with three, five, four and six patients enrolled, respectively. The MTD of paclitaxel was found at 50 mg/m2/wk and cisplatin 30 mg/m2/wk. Diarrhea was the dose-limiting toxicity. Thirteen patients were evaluable for response: seven complete and five partial responses were obtained with an overall response rate of 92.3%. CONCLUSIONS: The MTD of paclitaxel is 50 mg/m2/wk when associated to cisplatin 30 mg/m2/wk and concurrent pelvic radiotherapy. Diarrhea is the dose limiting side effect. Preliminary data suggest that concurrent chemoradiotherapy with paclitaxel and cisplatin could be a very active treatment for patients with locally advanced carcinoma of the cervix. PMID- 10847467 TI - Oxaliplatin and raltitrexed combined with leucovorin-modulated 5-fluorouracil i.v. bolus every two weeks: a dose finding study in advanced previously treated colorectal carcinoma. Southern Italy Cooperative Oncology Group. AB - PURPOSE: To determine the maximum tolerated dose of oxaliplatin (L-OHP) given as a two-hour infusion followed by raltitrexed (Tomudex [TOM]) administered as a 15 min infusion on day 1, and bolus 5-fluorouracil (5-FU) modulated by a fixed dose of levo-folinic acid (LFA) 250 mg/m2 on day 2, recycling every two weeks, and to have preliminary evidence of activity of this combination in pretreated advanced colorectal cancer patients. PATIENTS AND METHODS: Fifty-two patients with advanced colorectal carcinoma previously treated with one (25 cases) or two or more lines of chemotherapy, including irinotecan (26 cases), and/or modulated 5 FU (40 cases) entered this study. Starting doses of L-OHP, TOM, and 5-FU were 85, 2.5 and 750 mg/m2, respectively. RESULTS: Seven dose levels were tested. Neutropenia was the main dose limiting toxicity of the dose escalation (8 of 13 cases). The recommended doses were 130 mg/m2 of L-OHP, and 3.0 mg/m2 of TOM on day 1, followed by 250 mg/m2 of LFA, and 1050 mg/m2 of 5-FU on day 2, every two weeks. Severe diarrhoea and stomatitis were rarely reported. Most patients complained of mild peripheral sensitive aeurotoxicity, which was related to the cumulative dose of L-OHP. Twelve patients were considered as having a major responses (one complete), and an additional eight patients showed a minor response; the median time to treatment failure was twenty-four weeks. CONCLUSIONS: With this regimen it is possible to give full doses of all three cytotoxic drugs every two weeks. Its activity and its manageable toxicity profile deserve further evaluation in pretreated advanced colorectal cancer patients. PMID- 10847468 TI - Psychosocial predictors of survival: metastatic breast cancer. AB - BACKGROUND: Research interest in psychosocial predictors of the onset and course of cancer has been active since the 1950s. Recently we reported associations between psychological factors and survival in patients with metastatic melanoma. We now report a replication of this study in a sample of women with metastatic breast cancer. PATIENTS AND METHODS: Ninety-nine patients with metastatic breast cancer completed questionnaires measuring cognitive appraisal of threat, coping, psychological adjustment, perceived aim of treatment, social support and quality of life, approximately four months after diagnosis. Survival was measured from date of study entry to date of death or censored at the date of last follow-up for surviving patients. RESULTS: In a multivariate analysis, four factors independently predicted outcome. Patients with metastases in the liver, lung or pleura survived for a shorter duration (P < 0.001); older patients (P < 0.001) and those with a better appetite (P < 0.05) also lived for a shorter time. Patients who minimised the impact of cancer survived longer (a median of 29.1 vs. 23.9 months after study entry, P < 0.01). CONCLUSIONS: Minimisation was also significantly associated with outcome in patients with metastatic melanoma who participated in an identically designed study, reported elsewhere. This suggests that minimisation may have a general impact on cancer progression and deserves closer scrutiny in other cancers. PMID- 10847469 TI - Discrepancies in Epstein-Barr virus association at presentation and relapse of classical Hodgkin's disease: impact on pathogenesis. AB - BACKGROUND: Though Epstein-Barr virus (EBV) has been linked to classical Hodgkin's disease (cHD), only a proportion of cHD cases show EBV-association. While there has been evidence to suggest a crucial role for EBV in the early phase of cHD evolution, we are unclear whether continued presence of EBV is essential for the maintenance of the neoplasm. We have addressed this issue by investigating the EBV-association in paired samples of cHD obtained at presentation and relapse. MATERIALS AND METHODS: We have analysed lymph node biopsy samples from 23 cases of cHD at presentation and relapse. In situ hybridization with EBER-1 probe and immunostaining with anti latent membrane protein-1 (LMP-1) antibody was performed on the paraffin sections. PCR for EBNA 3C gene was performed for EBV subtype analysis on the DNA extracted from paraffin sections. RESULTS: EBV-association was documented in both the presentation and relapse samples of 14 cases. One case showed loss of EBV-association at relapse. PCR analysable DNA was obtained from both presentation and relapse samples in four of the EBV-associated cases. The EBNA 3C amplimer corresponded to type A strain of EBV in all the samples. CONCLUSION: Loss of EBV-association between presentation and relapse seen in one case implies a hit and run mechanism in EBV induced lymphomagenesis. Though EBV may be essential early in the evolution of cHD, it may not be required for maintenance of the neoplastic clone. Our study also brings out a speculation that a proportion of EBV-negative cHD could represent loss of EBV in the tumour prior to clinical presentation. Alternatively, an unidentified lymphotropic virus other than EBV might play a pathogenetic role in EBV-negative cHD. PMID- 10847470 TI - Gemcitabine-induced atrial fibrillation: a hitherto unreported manifestation of drug toxicity. AB - BACKGROUND: Gemcitabine is an antimetabolic drug for solid tumors. Although its pharmacokinetics as well as its side-effects are well known, paroxysmal atrial fibrillation associated to the administration of this drug has not yet been described. PATIENTS AND METHODS: We describe the case of a 78-year-old man with pancreatic adenocarcinoma who presented repeated paroxysmal atrial fibrillation episodes 18-24 hours after every gemcitabine infusion which resolved with antiarrhythmic drugs. This clinical history was positive for a remote brief episode of atrial fibrillation, which resolved spontaneously, and the patient had no predisposing factors for supraventricular arrhythmias (systemic hypertension, diabetes or coronary artery disease). RESULTS: Cardiac work-up revealed only a mild mitral-valve prolapse and complete right bundle branch block. During the arrhythmia episodes no other precipitating factors were reported. The close temporal relationship of the arrhythmia to drug administration and the recurrence of arrhythmia upon rechallenge allowed to hypothesize an intrinsic pro-arrhythmic effect of gemcitabine or its metabolite 2',2'-difluorodeoxyuridine. CONCLUSIONS: The occurrence of atrial fibrillation during the administration of gemcitabine may be considered as a cardiac arrhythmia drug-related toxicity. This side-effect of gemcitabine infusion is a previously unreported sign of drug toxicity; therefore, a high level of awareness to this problem is warranted when this drug is administered. PMID- 10847471 TI - Ten-year survival in advanced malignant melanoma following treatment with interferon and vindesine. AB - A 31-year-old man with malignant melanoma of his right popliteal fossa was treated in 1987 with surgical excision followed by local radiotherapy. Eight months later, he presented with recurrence in the right inguinal lymph nodes, which were resected and followed by radiotherapy to the groin. Ten months later, he developed liver metastases and was treated with vindesine (12 months) and interferon-alpha-2a (30 months) resulting in complete remission which has been maintained for over 10 years. This interesting case report, with brief review of literature, is presented here. PMID- 10847472 TI - Low doses interferon-alpha in the treatment of high-risk cutaneous melanoma. Melanoma Cooperative Group. PMID- 10847473 TI - Long-term octreotide treatment of metastatic carcinoid tumor. AB - The optimal dosage of somatostatin analogs for the long-term control of carcinoid tumors has not yet been established. Receptor alterations induced during long term treatment with somatostatin analogs have lead to consecutive drug dosage increases in order to control carcinoid disease. In this report, we describe the rapid and effective control of tumor in a patient with metastatic carcinoid treated for nine years with a single daily dose of octreotide based on tumor marker levels. Tumoral somatostatin receptor (sst) subtype analysis by RT-PCR amplification showed the expression of sst2 subtype only. We suggest that a single daily dose of octreotide strictly related to tumor marker secretion, could have played a role in the effective long-term therapy by avoiding the phenomenon of somatostatin receptor desensitisation. Furthermore, the exclusive presence of sst2 subtype supports the high affinity of octreotide to tumoral cells. PMID- 10847475 TI - Immunohemolytic anemia following oxaliplatin administration. PMID- 10847474 TI - Gemcitabine and vinorelbine in pretreated advanced breast cancer: a pilot study. AB - PURPOSE: Gemcitabine (GEM) and vinorelbine (VNR) are both active against advanced breast cancer (ABC), being able to induce a median ORR of 25% and 40%, respectively. Because of their different mechanism of action and good tolerability, the combination of GEM and VNR has been tested in ABC. PATIENTS AND METHODS: Twenty-nine ABC patients pretreated with anthracycline-taxane were treated with GEM 1000 mg/m2 on day 1, 8, 15, and VNR 25 mg/m2 on day 1 and 8 every twenty-eight days. Analysis of toxicity pattern, response rate, TTP and OS were carried out. RESULTS: Twenty-nine patients were enrolled into the trial. The ORR was 48% (95% CI: 29-67): a CR was observed in three patients (10%; 95% CI: 2 27), while eleven patients (38%; 95 CI: 21-58) achieved PR, eight (28%) had a SD, and seven (24%) progressed. Toxicity was mainly hematological and included: grade 3 leukopenia in 48% of cases without episodes of neutropenic fever, grade 3-4 thrombocytopenia in 10%, and grade 2 anemia in 7%. Non-hematological toxicities were mild and rather infrequent. CONCLUSIONS: The GEM-VNR combination seems to be active in pretreated ABC with an acceptable toxicity pattern, and may well reppresent an interesting therapeutic choice after anthracycline/taxane regimens. PMID- 10847476 TI - Liver transplantation for primary sclerosing cholangitis. AB - Primary sclerosing cholangitis (PSC) is a chronic cholestatic liver disease of unknown aetiology that is progressive in most symptomatic patients, advancing toward cirrhosis and liver failure. Liver transplantation is the only therapeutic option for patients with end stage liver disease resulting from this disorder. The results of transplantation for PSC are excellent with one-year survival rates of 90-97% and five-year survival rates of 80-85%, but are closely related to pre transplant Child-Pugh stage. Recurrence of PSC after liver transplantation is common, occurring in up to 20% of patients, but it appears to have little effect on patient survival, as survival of patients with recurrent PSC is similar to that of those without evidence of recurrence. Cholangiocarcinoma is a catastrophic complication of PSC and as yet no reliable screening method exists. The results of liver transplantation for patients with clinically apparent cholangiocarcinoma are extremely poor, however in patients in whom a microscopic tumour is detected in the explanted liver, survival is similar to those transplanted with PSC without cholangiocarcinoma. Activity of inflammatory bowel disease (IBD) appears to be more severe after transplantation, especially in units where steroid immunosuppression is withdrawn early. Colon cancer appears within the first few years after transplantation in approximately 7% of patients with IBD who are transplanted for PSC. Annual colonoscopy in this population seems prudent. PMID- 10847477 TI - Expression of gap junction protein connexin 32 in chronic liver diseases. AB - BACKGROUND: Gap junctions contain intercellular channels through which contacting cells communicate directly. The expression of connexin 32, a major gap junction protein in the liver, during the progression of chronic liver diseases has not yet been clarified. METHODS: Immunohistochemical staining was performed using anti-connexin 32 antibody on 6 specimens of normal human liver, 7 of chronic viral hepatitis, and 7 of liver cirrhosis. RESULTS: The number of gap junction plaques in chronic viral hepatitis and liver cirrhosis was significantly smaller than that in normal liver (10350+/-2180 and 7550+/-3040 vs 22560+/-3700 spots/mm2, p<0.01). The number of gap junction plaques tended to be lower in liver cirrhosis than in chronic viral hepatitis. CONCLUSION: These results suggest that in chronic liver diseases impaired intercellular communication between hepatocytes occurs. PMID- 10847478 TI - Growth inhibition of liver metastases by the anti-angiogenic drug TNP-470. AB - OBJECTIVE: This study was undertaken in order to assess the efficacy of a potent angiogenesis inhibitor, TNP-470, on tumor growth in a syngeneic rodent model of liver metastases from colorectal cancer. BACKGROUND: New blood vessel formation is a prerequisite for primary and metastatic tumor growth. TNP-470, a synthetic derivative of fumagillin when subcutaneously transplanted into nude mice, inhibits endothelial cell proliferation and migration, as well as the growth of various human cancers. However, the antitumor effect of this drug has not been studied in models reproducing a natural metastatic environment. Since the liver provides an extensive vascular bed for secondary tumor growth, an anti-angiogenic strategy may therefore be less efficient for treating hepatic metastases than primary tumors. METHODS: 10(7) DHD K12 colon carcinoma cells were injected intrasplenically into syngeneic BD IX rats to produce diffuse liver metastases. TNP-470 (30 mg/kg/day) was administered on alternate days starting 4 days after tumor implantation. The animals were sacrificed after 4 weeks and their livers were processed for histologic examination. In both the treatment and control groups (n=7), tumor volume was determined using a computerized analytical system, and tumor microvessel density was measured by immunostaining with anti-von Willebrand Factor monoclonal antibody. RESULTS: In vitro, TNP-470 demonstrated a direct toxicity towards the DHD K12 cell line with an IC50 of 0.1 microg/ml. Metastases were present in all animals from both groups. Liver weight (15.2 g vs 11.7 g, p=0.01), and tumor volume (1218 mm3 vs 406 mm3, p=0.03) were significantly reduced in the TNP-470 group compared to the control group. Tumor microvessel density was not statistically different between the two groups (67 vs 63 microvessels/x200 field, p=0.41). CONCLUSION: TNP-470 inhibits the growth of liver metastases in a syngeneic rat model of colorectal cancer. The mechanism responsible for this effect remains unclear, but may involve a combination of anti-angiogenic and direct cytotoxic effects. PMID- 10847479 TI - Acute obstructive cholangiopathy in interleukin-6 deficient mice: compensation by leukemia inhibitory factor (LIF) suggests importance of gp-130 signaling in the ductular reaction. AB - AIM: The hypothesis that interleukin-6-IL-6/gp130 signaling is involved in liver and biliary epithelial cell (BEC) biology and growth control was tested by subjecting homozygous IL-6 deficient mice (IL-6-/-) and wild type (IL-6+/+) littermate controls to bile duct ligation (BDL). MATERIALS AND METHODS: During the first week after BDL, the two groups were compared with respect to routine liver injury tests, liver histology, BEC and hepatocyte DNA synthesis, together with the expression of mRNA and protein of IL-6 as well as related growth factors, and their receptors. RESULTS: During the first week after BDL, there was marked upregulation of IL-6 mRNA and protein in the IL-6+/+ mice only in the vicinity of the biliary tree; whereas, biliary/peri-biliary IL-6R, HGF and met mRNA and protein increased in both groups. IL-6, HGF mRNA and protein localized to periductal inflammatory cells and stellate cells, while met and IL-6R protein were upregulated in the BEC and, to a lesser extent, in hepatocytes. This occurred during maximal proliferation of the BEC. Despite the absence of IL-6 in the IL-6-/- mice, there were only mildly phenotypic differences between the two groups, and no differences in mortality. Compared to IL-6+/+ controls, IL-6-/- mice showed slightly less BEC proliferation, a trend toward more liver injury, and significantly higher total serum bilirubin (TB) levels, suggestive of impaired biliary tree integrity. These changes were associated with slightly less HGF mRNA and protein expression in the IL-6-/- mice, but the differences were not significant. Leukemia inhibitory factor (LIF), another gp-130 ligand, also showed marked peri-biliary upregulation after BDL in both groups, and also induced BEC DNA synthesis, in vitro. CONCLUSIONS: The mild phenotypical differences between IL-6+/+ and IL-6-/- mice in the acute response to BDL is most likely attributable to the redundancy of the gp-130 signaling system. However, the long-term response to BDL results in a distinct phenotype in the IL-6-/- mice, marked by a relentless rise in serum total bilirubin and an inability to maintain compensatory increase in liver mass. PMID- 10847480 TI - Iron enhances hepatitis C virus replication in cultured human hepatocytes. AB - BACKGROUND: Iron overload in the presence of increasing concentrations of iron is one of the indicators of poor response to interferon therapy in chronic hepatitis C. In order to analyze the effect of iron on hepatitis C virus (HCV) replication, we measured replication in an HCV-infected cell line. METHODS AND RESULTS: Cells from a non-neoplastic HCV-infected human hepatocyte line (PH5CH8) susceptible to HCV infection and supportive of HCV replication were used in this study. The replication of HCV RNA was measured by reverse transcription-nested polymerase chain reaction (RT-nested PCR). PH5CH8 cell viability was measured by 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. PH5CH8 cells were incubated with 0, 1, 10, 50, and 100 microM of FeSO4 at 37 degrees C with 5% CO2. Forty-eight hours after iron supplementation, the quantity of HCV RNA in the cells incubated in 50 and 100 microM of FeSO4 was approximately ten times that of the cells with no iron supplementation. Similar changes were observed beginning at 12 h from supplementation with FeSO4 and continued for at least 72 h after supplementation. MTT assay indicated that iron did not have cytotoxic effects on the PH5CH8 cells. CONCLUSION: Iron enhances HCV replication in a hepatocyte cell line. The results suggest that iron deposition in hepatocytes could facilitate HCV infection in the liver. PMID- 10847481 TI - Fas ligand expressing mononuclear cells around intrahepatic bile ducts co-express CD68 in primary biliary cirrhosis. AB - AIMS/BACKGROUND: Apoptosis, including the Fas system, has been implicated in progressive bile duct loss in primary biliary cirrhosis (PBC). In this study, we attempted to analyze Fas ligand (FasL) expressing mononuclear cells infiltrating in the portal tracts of PBC. METHODS: We immunohistochemically assessed co expression of leukocyte markers and FasL on infiltrating mononuclear cells in 18 patients with PBC. Twenty-five patients with chronic hepatitis C (CH-C) were used as controls. RESULTS: In PBC, FasL expressing cells were scattered in the portal tracts, and some were accentuated around the damaged bile ducts. In addition, these cells co-expressed CD68 (71%), a marker of monocytes, but not UCHL-1, CD3 and CD57, markers of activated T cells and natural killer cells. By contrast, in CH-C, the biliocentric pattern of FasL expression was not evident, and about half of FasL expressing cells (42-56%) co-expressed UCHL-1, CD3 or CD57. CD14, a receptor for bacterial products such as lipopolysaccharides, was also detected on a proportion of FasL expressing mononuclear cells around the damaged bile ducts in PBC. CONCLUSION: The results suggest that in PBC, FasL expressing CD68+ monocytes are at least partly involved in apoptotic bile duct loss mediated by the Fas system, and a surface molecule, CD14, participates in this process. PMID- 10847482 TI - Genomic variations in precore and cytotoxic T lymphocyte regions in chronic hepatitis B in relationship to interferon responsiveness. AB - AIMS: Treatment of chronic hepatitis B patients with interferon (IFN) alpha results in a sustained loss of viral replication in as many as 20-40% of cases. The resistance to antiviral treatment by some viruses is due to positive selection of variants harboring mutations that confer resistance. An HBV variant with a stop codon in the precore region (A1896 HBV) has been reported to be associated with the response to IFN therapy. Recently variations in amino acid residues 21 and 27 of the core protein that serve as either partial agonists or antagonists to cytotoxic T-lymphocyte (CTL) function have been demonstrated to affect response to IFN therapy. In this study, we investigated whether these genomic variations in precore and CTL regions can affect response to IFN therapy. PATIENTS: Twenty-three patients with chronic hepatitis B were treated with IFN alpha. Of these 23 patients, 6 were responders (defined as showing seroconversion to anti-HBe, loss of serum HBV DNA and normalization of serum aminotransferase levels), and the remaining 17 were non-responders. METHOD: A total of 75 serum samples (3 serial samples per patient collected before, at the end of therapy, and 6 months after therapy) were tested for core promoter and precore wildtype and mutant viral levels and for sequence in the CTL region. RESULTS: There were no significant differences in precore wildtype and mutant viral levels before therapy between responders and non-responders, and both viral levels significantly decreased equally when measured at the end of therapy. Approximately 90% of non-responders with HLA-A2 had no amino acid substitutions in the CTL region before therapy. IFN therapy did not induce any specific mutations in this region. CONCLUSION: These findings suggest that the precore wildtype and mutant had similar sensitivity to IFN, and that genomic variation in the CTL region does not appear to be associated with response to IFN therapy. PMID- 10847483 TI - Telomerase activity correlates with cell cycle regulators in human hepatocellular carcinoma. AB - AIMS/BACKGROUND: Mutation in cell cycle genes is the most common genetic change in malignant tumor cells. Telomerase activation, considered as essential in the immortality of cancer cells, is found in most cancers, where there may be an association with an active cell cycle. METHODS: In this study study we used the TRAP assay to determine telomerase activity in liver tumor specimens from 25 cases of hepatocellular carcinoma (HCCs) as well as in corresponding non cancerous liver tissue in each patient. The expression of cyclin D1, cdk2, and cdk4 protein was also examined by Western blot. RESULTS: Twenty-one of the 25 cases of HCC were found to have increased telomerase activity, whereas only five out of the 25 non-cancerous liver samples were found to have weak telomerase activity. Telomerase activity was not found to be related to tumor size, HBsAg, HBeAg, anti-HCV, transaminase, or alpha-fetoprotein serum titer. Furthermore, three out of the 25 cases of HCC showed cyclin D1 overexpression, whereas 15 of the 23 cases of HCC showed decreased cyclin D1 expression. Down regulation of cyclin D1, cdk2, cdk4 protein correlated with telomerase activity (p<0.004, p<0.013, and p<0.001 respectively). CONCLUSION: The results indicate that genetic defects in HCC facilitate the reactivation of telomerase activity, a process which may be dependent on cyclin D1 with its cyclin dependent kinase (cdk) partner defect. PMID- 10847484 TI - Mast cell subpopulations in chronic inflammatory hepatobiliary diseases. AB - AIM/BACKGROUND: In various hepatobiliary diseases mast cells have been found to be associated with fibrogenesis. However, mast cell subpopulations have not been investigated in the human liver in normal subjects or in disease. Human mast cells are categorized into mast cells positive for tryptase (MC(T)) only and mast cells positive for both tryptase and chymase (MC(TC)). METHODS: In this study we investigated mast cell subpopulations (MC(T) and MC(TC)) by double immunostaining for mast cell tryptase and chymase as well as by a computer-aided quantitative morphometry in 13 normal livers and in 193 liver tissue specimens comprising of primary biliary cirrhosis (n=43), autoimmune hepatitis (n=11), chronic hepatitis B (n=37), chronic hepatitis C (n=41), alcoholic liver disease (n=40) and hepatolithiasis (n=21). RESULTS: The densities of MC(T) and MC(TC) per 1 mm2 stroma were low in normal livers but high in chronic liver diseases, and correlated positively with the degree of fibrosis. The percentages of MC(T) and MC(TC) subpopulations were 25% and 75%, respectively. The percentage was almost the same in normal livers and various hepatobliliary diseases, as well as between less fibrotic cases and more fibrotic cases in liver diseases. CONCLUSIONS: These results suggest that MC(T) and MC(TC) subpopulations in healthy and diseased livers do not change during liver fibrosis of any etiology. PMID- 10847486 TI - Cloning of the rat ADAMTS-1 gene and its down regulation in endothelial cells in cirrhotic rats. AB - AIMS: This study was undertaken in order to identify genes which are regulated during the process of liver fibrosis. METHODS: The differential display method and RNA from rat endothelial cells before and after induction of cirrhosis were used. RESULTS: A 496 bp fragment, which was down regulated in liver endothelial cells from a cirrhotic animal, was cloned. The cloned fragment showed a 95% homology with the newly cloned mouse ADAMTS-1 gene (a disintegrin and metalloproteinase with thrombospondin motifs), which is implicated in inflammation. The fragment was found to span the 3' of exon 6, the whole exon 7 and the 5' of exon 8. Sequencing of the entire coding region of the rat gene showed a 94% homology at the nucleic acid level and 96% homology at the amino acid level. The sequences responsible for the function of the protein were conserved. Northern blot analysis, using the cloned fragment as a probe, confirmed the finding that the gene was down-regulated in endothelial cells derived from livers of cirrhotic animals. In situ PCR analysis localised the ADAMTS-1 gene in the liver endothelial cells from normal animals. CONCLUSIONS: Regulation of the expression of genes which belong to the metalloproteinase family in liver endothelial cells might be important in the development of liver cirrhosis. PMID- 10847485 TI - Characterization of hepatocyte-derived mitogenic activity on hepatic stellate cells. AB - BACKGROUND/AIMS: Hepatic stellate cells (HSC) are located in close proximity to hepatocytes in Disse's space. Hepatocyte derived factors have earlier been implicated in the paracrine regulation of HSC proliferation. The aim of the present study was to further characterize this mitogenic activity of the parenchymal cell conditioned medium (PCcM). METHODS: Primary rat HSC were cultured for 4 days. DNA synthesis was measured by 3H-thymidine incorporation. TGFbeta1 immunoreactivity was quantified by ELISA. PCcM was obtained from hepatocytes cultured in medium without serum or hormones for two days. RESULTS: Incubation of 4-day-old HSC on plastic surface with PCcM for 2 days increased DNA synthesis, while no effect was seen in HSC cultured on Matrigel. Heat-, acid-, and protease-treatment of PCcM abolished its stimulatory effect. Size fractionations with spin columns indicated that the stimulatory effect was contained in the fractions of a molecular size between 30 and 100 kD. The addition of LY 294002, a phosphatidylinositol 3-kinase (PI3-K) inhibitor, dose dependently inhibited the PCcM induced increase in DNA synthesis to about 9% of the control values. The specific MAP kinase (MAPK) inhibitor, PD 98059 only suppressed the PCcM induced DNA synthesis to 35% of control cultures at the highest dose (10 microM). DNA content in the cultures was not affected by either blocker. HSC seemed to produce immunoreactive TGFbeta1. However, addition of latency-associated peptide (LAP), a potent TGFbeta1 blocker, stimulated DNA synthesis to a much less extent than PCcM. CONCLUSIONS: The factor(s) that stimulate DNA synthesis in HSC from hepatocytes are most likely protein(s) with a molecular size between 30-100 kD. These factor(s) rely more on PI3-K than on MAPK for their mitogenic effect and are probably not acting via TGFbeta1 inhibition. PMID- 10847487 TI - Expression of cyclin D1, cyclin E, cdk4 and loss of heterozygosity of 8p, 13q, 17p in hepatocellular carcinoma: comparison study of childhood and adult hepatocellular carcinoma. AB - AIMS/BACKGROUND: In hepatitis B virus-related hepatocellular carcinoma (HCC), at least 20-40 years of continuous necro-inflammation is necessary for the hepato carcinogenesis to occur. However, HCC in childhood shows an unusually short latent period and rapid progression. In our previous report, mutation of c-met was found only in childhood HCC, but not in adult HCC. In order to determine the specific biological tumorous features of childhood HCC, a comparison study of childhood and adult HCC was performed. METHODS: Eighteen cases of HBV positive HCC (nine children and nine adults aged more than 40 years) were selected. The expression of G1 phase regulatory proteins (cyclin D1, cyclin E and cdk4) was studied using immunohistochemical methods. Loss of heterozygosity (LOH) on chromosomal arms 8p, 13q and 17p was analyzed. RESULTS: Cyclin D1 expression was significantly lower in childhood HCC than in adult HCC (cases of cyclin D1 expression under 3+: childhood 5/9 vs. adult 1/9, p=0.046). No difference in cyclin E and cdk4 expression was found between childhood and adult HCC. LOH frequency on 13q was relatively higher in childhood than in adult HCC (66.7% vs. 22.2%, p=0.058). LOH frequency on 8p and 17p was 44.4% and 33.3% in childhood HCC and 44.4% and 75% in adult HCC with no statistical significance between the two groups. CONCLUSION: Our data suggest that childhood HCC may be less dependent on cyclin D1 protein for tumor growth and progression than adult HCC, and that the LOH on 13q may be an important feature of childhood HCC. PMID- 10847488 TI - Continuous infusion of prostaglandin E1 via the superior mesenteric artery can prevent hepatic injury in hepatic artery interruption through passive portal oxygenation. AB - AIMS/BACKGROUND: Hepatic artery interruption (HAI) causes severe ischemic liver damage, especially following hepatopancreatobiliary surgery. In order to inhibit a decrease in oxygen delivery after HAI, continuous infusion of PGE1 via the superior mesenteric artery (SMA) was administered in pigs and changes in hepatic blood flow and oxygen delivery were investigated. Furthermore, its effectiveness in the prevention of liver injury was evaluated by histology and serum enzyme levels. METHODS: Animals were subjected to HAI without PGE1 infusion (control group n=6) and to continuous infusion of PGE1 (0.02 microg/kg/min) into the SMA (PGE1 group n=6). RESULTS AND CONCLUSION: PGE1 infusion via the SMA not only increased the portal blood flow but also elevated the oxygen content of the portal blood. The reduction in oxygen delivery to the liver was 50% in the control group, and only 13% in the PGE1 group. Serum aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) levels 24 h after HAI in the control group were 3415+/-1283 IU/L and 9839+/-2959 respectively while in the PGE1 group they were 939+/-426 IU/L and 5510+/-1545 IU/L respectively. Histological examination showed massive necrosis in the control group at 72 h but only focal liver cell necrosis in the PGE1 group. Based on this finding and the fact that this treatment can be performed easily and safely, continuous infusion of PGE1 via the SMA may be a useful intervention to prevent severe liver damage after hepatic artery interruption. PMID- 10847489 TI - Hepatocellular carcinoma presenting with right supraclavicular lymph node metastasis and superior mediastinal syndrome. PMID- 10847490 TI - Management of hepatocellular adenoma during pregnancy. PMID- 10847491 TI - Metachronous viral hepatitis. PMID- 10847492 TI - Optical coherence tomography for retinal thickness measurement in diabetic patients without clinically significant macular edema. AB - BACKGROUND AND OBJECTIVE: To evaluate the potential of optical coherence tomography (OCT) as a screening method for retinal thickness measurements in diabetic patients. PATIENTS AND METHODS: We used a previously described pattern of six 5 mm OCT scans through the center of fixation in 45 diabetic patients without clinically significant macular edema: 22 patients (group 1) had no diabetic retinopathy (ETDRS classification); 18 (group 2) had nonproliferative retinopathy; 5 patients with peripheral neovascularization did not enter statistical analysis; 25 normal healthy subjects were used as a control group. Retinal thickness was measured at five locations in each scan: in the fovea, at the foveal rim, and outside the macula. Measurements were identified in nonaligned images and taken from raw data A-scans. Locations were grouped into hemispheres, quadrants and rings, and mean values tested for statistically significant differences using Mann-Whitney U-Wilcoxon rank sum W test. RESULTS: Differences in retinal thickness were found to be significant in the macula (controls vs group 2 P = 0.0266), at the foveal rim (controls vs group 1 and 2: P = 0.0386 and P = 0.0193), in the nasal and superior hemisphere (controls vs group 2: P = 0.0251 and P = 0.0187), and in the superior nasal quadrant (controls vs group 1 and group 1 vs group 2: P = 0.0022 and P = 0.0462). CONCLUSIONS: Significant differences of retinal thickness between patients with diabetic retinopathy and normals can be detected by OCT even in the absence of clinically significant macular edema. Significant differences between diabetic patients with and without retinopathy are most likely to be found in the superior nasal quadrant. PMID- 10847493 TI - Optical coherence tomography findings in diabetic macular edema before and after vitrectomy. AB - BACKGROUND AND OBJECTIVE: To assess the role of optical coherence tomography (OCT) in the evaluation and follow-up after vitrectomy for diabetic macular edema. MATERIALS AND METHODS: The 18 eyes of 12 patients affected by presumed diabetic vitreous-induced macular edema underwent biomicroscopy with a Goldmann contact lens, fluorescein angiography, and OCT. RESULTS: OCT revealed two patterns of edema. The first group (15 eyes) was characterized by widespread thickening of the neurosensory retina with an increased nonhomogeneous reflectivity of the inner retinal layers; cystoid-like spaces of absent or reduced reflectivity in the neurosensory retina were also present. In the second group (3 eyes), a cystoid macular edema with a dome-shaped foveal profile because of a markedly increased retinal thickness in the foveal region was observed. The disappearance of the physiologic foveal profile was always seen. Biomicroscopy revealed an increased reflex of the inner limiting membrane in the first group and minimal alterations in the second one. Four patients (7 eyes) underwent vitrectomy with posterior hyaloid removal. In 5 eyes, the patients experienced a visual improvement greater than two Snellen lines. The restoration of the normal foveal profile and the reduction of the retinal thickness on the OCT were evident in all cases one month after surgery. CONCLUSION: OCT appears to be a useful tool in the diagnosis and management of diabetic macular edema and in the monitoring of the morphological changes after vitrectomy. PMID- 10847494 TI - Optical coherence tomography after laser photocoagulation for clinically significant macular edema. AB - BACKGROUND AND OBJECTIVE: To demonstrate the utility of optical coherence tomography (OCT) for documenting an early response to laser photocoagulation in clinically significant macular edema (CSME) secondary to diabetes. PATIENTS AND METHODS: Five eyes of four patients were selected for review based on the diagnosis of CSME. All eyes had a clinical diagnosis of CSME based on slit lamp biomicroscopy. All eyes underwent focal/grid laser photocoagulation to areas of retinal thickening detected by OCT and clinical exam. Pre and post-treatment optical coherence tomograms were obtained for all patients. RESULTS: All eyes selected for review had an early positive response to focal laser photocoagulation. OCT was useful for demonstrating areas of retinal thickening prior to laser treatment. Serial macular maps demonstrated the resolution of retinal thickening after laser photocoagulation in all eyes. CONCLUSION: OCT is a useful tool for evaluating and documenting CSME both before and after focal/grid laser photocoagulation. OCT is capable of detecting an early positive response to photocoagulation for macular edema. PMID- 10847495 TI - Retina tomography after vitrectomy for macular edema of central retinal vein occlusion. AB - PURPOSE AND OBJECTIVE: To report retina tomography after vitrectomy for macular edema in central retinal vein occlusion (CRVO). PATIENTS AND METHODS: Five patients with macular edema caused by CRVO were examined using optical coherence tomography (OCT) through their clinical courses. RESULTS: The retinal thickness through the fixation was reduced in all 5 eyes. Preoperatively, the retina thickness at the foveola was more than 500 microm in three eyes and more than 1000 microm in 2 eyes. The retina thickness was reduced to 311+80 microm within two weeks on average. Visual acuity was improved by two or more lines in 3 of 5 eyes. CONCLUSION: Vitrectomy is worthy of consideration when macular edemas are prolonged in patients of CRVO. OCT is a useful instrument for management and treatment of macular edema. PMID- 10847496 TI - Dome-shaped detachment of premacular vitreous cortex in macular hole development. AB - PURPOSE: To clarify the role of the posterior vitreous cortex in macular hole development, we evaluated the follow-up findings of clinical observations and optical coherence tomography (OCT) images of macular holes in early stage. METHODS: The vitreoretinal tomography was examined and followed in eyes with stage 1 and 2 macular holes with OCT. RESULTS: The posterior cortical vitreous was identified in 12 of 17 eyes with early-stage macular holes by OCT (71%). In these eyes, the detached posterior vitreous appeared to be merged to the fovea and the disc. The vitreoretinal separations showed typically dome-shaped. A long scan along the papillomacular axis also delineated the posterior vitreous face as dome-shaped perifoveal vitreous detachment with adhesion to the fovea and disc. Repetitive OCT examinations clearly demonstrated the course of development of the macular hole. CONCLUSIONS: The dome-shaped vitreoretinal separation seen with OCT gives rise to an idea that the posterior vitreous cortex may not be taut but slack, and would not cause a continuous tractional force even in early-stage macular holes. This finding brings us an idea that the tangential traction at the fovea may be generated by fluid movements, rather than by contraction of the cortical vitreous. PMID- 10847497 TI - Optical coherence tomographic findings of idiopathic polypoidal choroidal vasculopathy. AB - BACKGROUND AND OBJECTIVE: To identify the histological level of abnormal vessels associated with idiopathic polypoidal choroidal vasculopathy (IPCV), we examined IPCV with Optical Coherence Tomography (OCT). PATIENTS AND METHODS: Fourteen patients diagnosed with IPCV were examined with Indocyanine green (ICG) angiography and OCT. RESULTS: ICG angiography demonstrated branching vascular networks with polypoidal dilatations at the terminals beneath the retinal pigment epithelium (RPE). OCT showed dome-like elevation of the RPE, and moderate reflex or nodular appearance were seen beneath the RPE. CONCLUSION: The abnormal vessel associated with IPCV is supposed to be choroidal neovascularization with polypoidal dilatations at the terminals between Bruch's membrane and RPE. We consider that this disease is a peculiar form of age-related macular degeneration. PMID- 10847498 TI - Retinal thickness immediately after cataract surgery measured by optical coherence tomography. AB - BACKGROUND AND OBJECTIVE: To evaluate changes in retinal thickness immediately after cataract surgery. PATIENTS AND METHODS: Retinal thickness before and 0.5 hours after uneventful cataract surgery was determined in 10 patients at 15 different points at the posterior pole by optical coherence tomography (OCT). RESULTS: The mean retinal thickness at the posterior pole was 251 microm (+/-30 microm) before cataract surgery and 249 microm (+/-25 microm) after cataract surgery. No statistical difference between pre- and postoperatively values could be evaluated (P = 0.8). CONCLUSIONS: The results indicate that uncomplicated cataract surgery does not influence retinal thickness immediately postoperatively in eyes without ocular pathologies. PMID- 10847499 TI - First experimental and clinical results with transscleral optical coherence tomography. AB - BACKGROUND AND OBJECTIVE: To evaluate the potentials of optical coherence tomagraphy (OCT) using long wavelength to penetrate highly scattering tissues of the eye and visualize the anterior chamber angle and the ciliary body. METHODS: OCT images were generated by an experimental prototype in enucleated porcine eyes using as light source a superluminiscent diode with a wavelength of 1310 nm and a scan frequency of 60 Hz. The number of lateral scans was variable in a range from 100 to 400. RESULTS: Infrared OCT was able to penetrate the sclera. The anterior chamber angle could be visualized completely and the ciliary body could be identified. However, it was not possible to penetrate the highly reflective iris pigment epithelium. CONCLUSION: The use of infrared OCT allows penetration of the sclera, thus, providing complete visualization of the anterior chamber angle and limited demonstration of the ciliary body. Because of its higher resolution, it may represent an interesting noninvasive alternative to ultrasound biomicroscopy. PMID- 10847500 TI - Grayscale and proportion-corrected optical coherence tomography images. AB - BACKGROUND AND OBJECTIVE: The commercially available optical coherence tomography (OCT) scanner displays images in a pre-set window regardless of the projected scan length on the retinal surface. The aim of this study was to demonstrate the true dimensions of proportion-corrected OCT images and the additional information present in grayscale images. MATERIALS AND METHODS: OCT raw data were exported to an IBM-compatible PC and processed to show grayscale and proportion-corrected images using an automated software of our own design. RESULTS: Eyes with cystoid macular edema and retinal pigment detachment were analyzed. Grayscale images showed a finer gradation of signal reflectance. Scan lengths of 2, 4, 6, and 8 mm on the retinal surface showed different qualitative appearances using proportion corrected software from the printed or on-screen images. CONCLUSIONS: Grayscale OCT images can be used to demonstrate additional information not present in false color images. The disparity between the standard OCT image format and proportion corrected images emphasizes the need for quantitative rather than qualitative evaluation of retinal dimensions and internal reflectance. PMID- 10847501 TI - Unilateral macular retinoschisis with stellate foveal appearance in two females with myopia. AB - Stellate foveal retinoschisis is considered the hallmark of X-linked retinoschisis. We describe the finding of unilateral retinoschisis with stellate foveal appearance in 2 females with myopia who had no evidence of familial disease. Optical coherence tomography (OCT) and fluorescein angiography were obtained in both cases. Neither patient had a family history of low vision, night blindness, or retinal detachment. Visual acuity in the affected eyes was 20/50 and 20/25, respectively. Both fellow eyes had normal fundi, except for mild myopic changes. With OCT, widespread retinal splitting was detected at the posterior pole in the affected eyes, whereas the fellow eyes were normal. In one case, OCT showed that foveal retinoschisis represented the macular involvement of a flat inferior retinoschisis, although this was not clinically apparent. Macular retinoschisis with stellate foveal appearance may rarely be associated with pathologic myopia. OCT was useful to establish the true extension of these macular changes. PMID- 10847502 TI - Reattachment of retina and retinoschisis in pit-macular syndrome by surgically induced vitreous detachment and gas tamponade. AB - The origin of the subretinal fluid in pit-macular syndrome is unknown. Using optical coherence tomography, the authors observed that an optic disk pit is not a true pit but a cystic cavity covered with a superficial layer of the optic disk in a patient with retinal detachment and retinoschisis. The cyst was connected to the intraretinal space of the retina, which was split in several layers, or the retinoschisis in the papillomacular area. Vitrectomy and gas tamponade were performed to treat the patient. During surgically-induced vitreous detachment, strong vitreoretinal attachment at the disk margin and the fovea was observed. After surgery, the cystic space at the optic disk disappeared and the retinal detachment and retinoschisis reattached. Vitreous traction may play a role to introduce the fluid from the optic cyst to the subretinal space through the superficially split retina. PMID- 10847503 TI - OCT in successful surgery of retinal detachment associated with optic nerve head pit. AB - The authors describe the course of a patient with retinal detachment associated with optic nerve head pit for whom optical coherence tomography (OCT) was performed before and after vitrectomy and gas tamponade. An 18-year-old woman presented with macular elevation and a lamellar macular hole associated with optic nerve head pit. OCT showed that the retinal elevation consisted of both separation of the inner and outer retinal layers and detachment of the outer layers from the retinal pigment epithelium. After vitreous surgery and postoperative intraocular gas tamponade, the macular elevation gradually resolved, and OCT showed that improvement in central vision corresponded with flattening of the outer layer detachment in the fovea. OCT is a useful tool for monitoring the therapeutic effect of vitrectomy and gas tamponade for retinal detachment associated with optic nerve head pit. PMID- 10847504 TI - Retinal detachment and retinoschisis detected by optical coherence tomography in a myopic eye with a macular hole. AB - The authors describe a myopic patient with a full-thickness macular hole that was accompanied by both retinal detachment and retinoschisis. A 51-year-old woman presented with a localized retinal elevation of three disc diameters around the macular hole in a myopic eye. Optical coherence tomography (OCT) showed features of both retinal detachment and retinoschisis. After vitreous surgery and postoperative intraocular gas tamponade, the macular elevation resolved, and separation of the neurosensory retina from the retinal pigment epithelium band disappeared on the OCT images. OCT is a useful tool for examining macular conditions associated with a macular hole in myopic patients. PMID- 10847505 TI - Confirmation of persistent closure of surgically repaired macular hole in subsequent retinal detachment by optical coherence tomography. AB - Idiopathic macular holes have shown response to surgical intervention with vitrectomy, presumably because of glial cell proliferation in the reapproximated tissue. We describe a patient with postoperative closure of a macular hole who subsequently developed a retinal detachment involving the macular area. Despite the detachment, the macular hole remained closed as evidenced by biomicroscopy and optical coherence tomography (OCT). OCT provides accurate documentation and assessment of pre- and postoperative conditions in macular hole surgery. PMID- 10847506 TI - Spontaneous closure of macular hole after posterior vitreous detachment. AB - Macular holes can be treated with surgically-induced vitreous detachment and gas tamponade. The authors report a case of a macular hole that closed spontaneously after the development of posterior vitreous detachment (PVD). Optical coherence tomography (OCT) revealed a small full-thickness macular hole with perifoveal cystic changes in a 25-year-old female with a central scotoma at her initial visit. There was no evidence of PVD. Five months later, PVD was observed and the macular hole disappeared. OCT performed three months later showed macular hole closure and resolution of the perifoveal cystic changes. The physiologic pit was restored in the fovea. OCT is useful to detect and monitor the morphologic changes of a macular hole. PMID- 10847507 TI - Occult retinal pigment epithelial detachment in hyperviscosity syndrome. AB - We document and evaluate a serous retinal detachment in a patient with hyperviscocity syndrome. Optical coherence tomographic images of the serous retinal detachment in a patient with hyperviscocity syndrome were correlated with slit-lamp biomicroscopic findings, fundus photographs, fluorescein angiograms, and indocyanine green angiograms. Fluorescein angiography demonstrated venous and capillary bed abnormalities but no leakage or pooling of fluorescein corresponding to the retinal pigment epithelial detachment (PED) beneath the serous retinal detachment. Indocyanine green angiogram disclosed a delay of intrachoroidal circulation. Optical coherence tomography (OCT) revealed a large retinal pigment epithelial detachment beneath the serous retinal detachment. The occult retinal PED beneath the neurosensory retinal detachment was detected only by OCT in a patient with hyperviscosity syndrome. We suggest that gamma globulin, which is the hyperviscosity material, accumulated in the subretinal pigment epithelial space and blocked the leakage or pooling of fluorescein corresponding to the retinal pigment epithelial detachment. PMID- 10847508 TI - Optical coherence tomography images of retinitis pigmentosa. AB - The retinas of 3 patients with typical retinitis pigmentosa (RP) were evaluated by optical coherence tomography (OCT), fluorescein angiography (FA), and fundus photography (FP). OCT showed that retinal thickness was decreased in the areas of the retina affected by RP, and nearly normal in the unaffected macular area. Reflectivity was partially decreased in the affected retinal areas and pigment masses were hyper-reflective. Reflectivity of the macular area was nearly normal except in the fovea, where it was decreased because of edema. Cystic spaces of the fovea were evident in Patient 1. PMID- 10847509 TI - A bibliography of optical coherence tomography basic science and ophthalmic applications. PMID- 10847510 TI - Reg Cooper. PMID- 10847511 TI - Hypothesis based research: the maturation of orthopaedic science. PMID- 10847512 TI - The role of ultrasonography in thromboembolic disease management in the orthopaedic patient. AB - Deep venous thrombosis (DVT) is a well-recognized contributor to increased morbidity and mortality following trauma and elective musculoskeletal procedures. Ultrasound has become a popular noninvasive modality for use in the detection of symptomatic DVT. However, its use as a screening tool in asymptomatic or postoperative patients has been questioned. The reliability of ultrasound rests mainly in the ability of the technicians performing the exam. Ultrasound has been shown to be less reliable in identifying asymptomatic calf thrombi; in institutions where ultrasound DVT surveillance is not performed routinely, the technique suffers from inadequate sensitivity to be utilized for routine screening purposes. Recognition of patients at high risk for DVT, along with an understanding of the limitations of ultrasound, will allow for appropriate clinical application of this modality. PMID- 10847513 TI - Does pulsed low intensity ultrasound allow early return to normal activities when treating stress fractures? A review of one tarsal navicular and eight tibial stress fractures. AB - We sought to evaluate the efficacy of daily pulsed low intensity ultrasound (LIUS) with early return to activities for the treatment of lower extremity stress fractures. Eight patients (2 males, 6 females) with radiographic and bone scan confirmed tibial stress fractures participated in this study. Additionally, a case report of a tarsal navicular stress fracture is described. All patients except one were involved in athletics. Prior to the study, subjects completed a 5 question, 10 cm visual analog scale (VAS) regarding pain level (10 = extreme pain, 1 = no pain) and were assessed for functional performance. Subjects received 20-minute LIUS treatments 5 times a week for 4 weeks. Subjects maintained all functional activities during the treatment period. Seven patients with posterior-medial stress fractures participated without a brace. Subjects were re-tested after 4 weeks of treatment. Mann-Whitney U tests (VAS data) and paired t-tests (functional tests) assessed statistical significance (p<0.05). Although the intensity of practice was diminished in some instances, no time off from competitive sports was prescribed for the patients with the tibial stress fractures. The patient with the anterior tibial stress fracture underwent tibial intramedullary nailing at the conclusion of a season of play. In this uncontrolled experience, treatment of tibial stress fractures with daily pulsed LIUS was effective in pain relief and early return to vigorous activity without bracing for the patients with posterior-medial stress fractures. PMID- 10847514 TI - Long-term follow-up of persistent humeral shaft non-unions treated with tricortical bone grafting and compression plating. AB - Three percent of all fractures occur in the humeral shaft. A very high union rate is achieved with nonoperative treatment. When non-unions occur, however, they are often very difficult to treat, and often require multiple procedures to achieve union. Even with multiple procedures, true pseudoarthroses have only a 59% union rate10. We conducted a retrospective study of ten patients having persistent non union of the humeral shaft, three of whom had a true pseudoarthrosis. An patients were treated with a compression plate and a tricortical iliac crest bone graft anchored rigidly across the fracture site with two screws applied at 90 degrees to the plate. A solid union was achieved in all ten (100%) patients. PMID- 10847515 TI - External fixation of open humerus fractures. AB - Fifteen patients with open shaft of humerus fractures were treated with a monolateral external fixator. Nine patients presented with nerve palsies. Two radial nerves were disrupted and required grafting. Of the seven others, six spontaneously recovered and one brachial plexus partially improved. All fractures healed. The average duration of external fixation was 21 weeks. Four patients required additional procedures prior to healing (external fixator reapplication 2, plating and bone grafting-2). Two of these four experienced breakage of 4.5 mm external fixation pins. Eight patients developed pin tract infections, which all resolved with local care and antibiotics. Thirteen patients were contacted at an average of 63 months after injury. Eleven reported they were satisfied with their result, nine had no functional limits, and eight reported no pain. PMID- 10847516 TI - Osteoporosis: the increasing role of the orthopaedist. AB - Osteoporosis is an ever-increasing problem as our population ages. However, it is also to a large extent a preventable problem. The orthopaedist now has the ability to determine bone mass, the rate of turnover, and the fracture risk. Skeletal bone mass can be evaluated with DXA; the rate of bone resorption can be determined by assessment of collagen-degradation urinary products; and the weight status, fracture history, and history of smoking can be used to predict the fracture risk in individual patients. The orthopaedic physician also needs to take an active role in advising their younger patients about achieving peak bone mass. All individuals should follow a program that includes adequate calcium replacement, 400 to 800 units of vitamin D, appropriate exercise, avoidance of significant weight loss, and cessation of smoking. At menopause, women should evaluate their risk factors and consider the use of estrogen not only for its skeletal benefits but also for its nonosseous effects. In patients with contraindications or an aversion to hormone therapy, bone densitometry should be performed to determine risks before expensive nonhormonal treatment is initiated. Additional studies such as measurement of collagen degradation products will help establish whether the patient's resorptive rate is high or stable. If the bone mass is 2.5 SDs below normal peak or if there is an increase in resorption, use of either estrogen, bisphosphontes, or calcitonin may be appropriate. If there is evidence of low-turnover osteoporosis with decreased osteoblast formation, sodium fluoride should be considered. Two thirds of the cost of osteoporosis in the United States is due to hip fractures. The orthopaedist is the primary physician who comes in contact with these fracture patients. It is therefore his or her responsibility to become knowledgeable about the treatment and prevention of osteoporosis. The bisphosphonates, hormones, and calcitonin provide predictable restoration of bone mass and significantly decrease the rate of osteoporotic fractures. PMID- 10847518 TI - Collagen fibril diameter distributions in rabbit anterior cruciate and medial collateral ligaments: changes with maturation. AB - This study presents morphometric analyses of the collagen fibril diameters of rabbit anterior cruciate and medial collateral knee ligaments of New Zealand White rabbits (young, age two months and adult, age thirty-six to forty months). Measurements were made from transmission electron micrographs of transverse ligament sections of approximately 50,000x magnification. Statistically significant differences in the mean fibril diameters were found between the anterior cruciate and medial collateral ligaments of the thirty-six to forty month old animals (.069 +/- .005, .092 +/- .016 mm, p < .1); however, no statistical significance was found for differences between these ligaments in two month old animals (.077 +/- .006, .082 +/- .009, p > .1). These data support the idea that known differences in fibril distributions of adult rabbit anterior cruciate and medial collateral ligaments develop with maturation, and may reflect both the cellular environment in which the fibrocytes of these ligaments are subject to, as well as the developmental genetic program of these cell populations. PMID- 10847517 TI - Pseudoachondroplastic dysplasia: an Iowa review from human to mouse. AB - Lamellar inclusions of the rough endoplasmic reticulum in growth plate chondrocytes, first identified (1972) in the Department of Orthopaedic Surgery, University of Iowa, has become the cytochemical hallmark for the pseudoachondroplastic dysplasia (PSACH) phenotype, linking an endoplasmic reticulum storage disorder with the osteochondrodysplasia. Since this original observation, great advances have been made, leading to the molecular understanding of this altered longitudinal bone growth anomaly. A PSACH canine model suggested that abatement of cumulative vertical growth of growth plate chondrocytes seen in PSACH results from (1) altered extracellular matrix constraints for horizontal growth and (2) uncoupling of endochondral and perichondral growth that causes metaphyseal flaring. PSACH, an autosomal dominant disease, is linked to mutation of the cartilage oligomeric matrix protein (COMP) gene. Amino acid substitutions, deletions, or additions is proposed to alter COMP structure that cause its retention in the rough endoplasmic reticulum of growth plate chondrocytes, leading to (1) compositional and structural change of the extracellular matrix, and (2) altered cellular proliferation and volume expansion. Normal growth and development occurs in COMP gene knockout mice that do not synthesis COMP, demonstrating that a mutant COMP, not absence of COMP, is required for the PSACH phenotype. The mechanism by which mutant COMP induces a PSACH phenotype remains to be elucidated. At the University of Iowa a cell culture system has been developed whereby mutant COMP transgenes are introduced into chondrocytes and the expressed product COMP is retained in the endoplasmic reticulum. This readily manipulated system makes it possible to decipher systematically the system's cellular secretory processing pathway, in order to clarify the mechanism(s) by which the mutant COMP is retained within the endoplasmic reticulum. Concurrent with this is the development of transgenic mice expressing the mutant COMP used in the cell culture system. This will make it possible to establish that expression of a human PSACH-linked mutant COMP will produce a PSACH phenotype. A PSACH animal model will provide a means to characterize the mechanism of altered longitudinal bone growth and to test gene therapy approaches for correcting the anomaly. PMID- 10847519 TI - Effect of intermittent administration of parathyroid hormone on fracture healing in ovariectomized rats. AB - To evaluate the potential use as a therapeutic agent for osteoporotic fractures, we examined the effects of intermittent administration of parathyroid hormone on fracture healing in ovariectomized rats. At three months post-ovariectomy, bilateral tibial shaft fractures were induced and stabilized by intramedullary nailing with Kirschnerwires. Saline, 17-estradiol, or recombinant human PTH(1-84) was given once a day for 30 consecutive days during fracture healing. Fracture healing was assessed by morphometric and mechanical analysis of fracture callus. Intermittent parathyroid hormone administration increased the morphometric and mechanical parameters in a dose-dependent manner. 17-estradiol, a bone-resorption inhibiting agent, did not offer advantage in terms of fracture healing in ovariectomized rats. Our findings suggest that intermittent parathyroid hormone administration may benefit osteoporosis and fracture. PMID- 10847520 TI - Hip osteotomy arthroplasty: ten-year follow-up. AB - We previously reported the initial success of combined osteotomy and arthroplasty of the hip for arthritis with femoral deformity. This technique has gained acceptance. We now report, for the first time, the ten year clinical and radiographic results with histology of 2 specimen. The osteotomies healed and the proximal femoral segment remained viable. One of three patients is symptom free without subsequent operative treatment. One of three patients had revision for acetabular loosening at eight years and biopsy of the proximal femur showed the proximal femoral segment to be viable. One of three patients had loosening of a macrofit bipolar prosthesis which required revision to total hip replacement at five years. Histology revealed viability of the proximal femur. All three patients are doing well at ten year follow-up. Based on the results of this study and current knowledge, the technique of osteotomy and arthroplasty for hip arthritis associated with femoral deformity is effective when combined with current techniques of ingrowth femoral component of total hip arthroplasty. PMID- 10847522 TI - Arthroscopic Mumford procedure variation of technique. AB - Fifty-seven patients had arthroscopic Mumford procedures for acromioclavicular pain non-responsive to conservative treatment. Thirty-nine of these patients had concomitant rotator cuff repairs. All had significant improvement of their distal clavicular pain. Neither the amount nor the completeness of distal clavicle resection affected the results. Arthroscopic distal clavicle resection is a safe and effective method of alleviating acromioclavicular pain. PMID- 10847521 TI - Posterior cruciate ligament function following total knee arthroplasty: the effect of joint line elevation. AB - One of the most commonly cited reasons for retaining the posterior cruciate ligament (PCL) during total knee arthroplasty is to preserve femoral rollback and theoretically improve extensor mechanism efficiency (lengthening the moment arm). This study was undertaken to assess PCL function in this regard and to delineate the effects of joint line elevation that can be manipulated intraoperatively by the surgeon. The anterior movement of tibiofemoral contact following PCL resection at flexion angles 60 degrees demonstrated the beneficial effect of the PCL on extensor function. This anterior translation and the concomitant increases in quadriceps tendon load and patellofemoral contact pressures were consistently observed. This study demonstrated that small changes of the joint line position significantly influenced PCL strain and knee kinematics. In order to preserve the desired functions that would be lost with an overly lax PCL and to avoid the potential adverse effects of an overly tight PCL (posterior edge loading and increased tibiofemoral contact), the surgeon should make every effort to restore the preoperative joint line. If this is not possible, consideration should be given to posterior cruciate recession or use of a posterior cruciate substituting design. PMID- 10847523 TI - The variable presentation and natural history of Langerhans cell histiocytosis. AB - Langerhans cell histiocytosis is not a well defined or predictable disease. Instead, it is a spectrum of disorders of unknown etiology that vary widely in presentation and natural history, but have in common the proliferation of histiocytic cells and infiltration of these cells into normal tissues. Although the lesions of Langerhans cell histiocytosis consist primarily of histiocytes, eosinophils are a prominent feature in some lesions. Lesions may develop in any tissue, but bone, skin and lymph nodes are the most commonly affected, and more than 75% of patients have skeletal lesions. Bone lesions caused by Langerhans cell histiocytosis vary from focal sharply defined areas of bone lysis to diffuse osteopenia and can resemble lesions caused by a wide variety of metabolic, infectious and neoplastic diseases. The natural history varies from a benign disorder that resolves spontaneously to a progressive fatal disease. In general, the younger the individual at the time of onset of the disease, the poorer the prognosis and the more extensive the disease. Treatment may include surgery, chemotherapy and radiation therapy, depending on the extent and severity of the disease. PMID- 10847524 TI - A brief history of workers' compensation. PMID- 10847526 TI - The diagnosis of the os trigonum syndrome with a fluoroscopically controlled injection of local anesthetic. AB - PURPOSE: To report the results of excision of the os trigonum using a fluoroscopically controlled injection of local anesthetic to diagnose the os trigonum syndrome. DESIGN AND PATIENTS: Os trigonum syndrome is a recognized cause of pain in the posterior aspect of the foot and ankle. The symptoms and physical findings, however, are often nonspecific and difficult to differentiate from other causes of posterior ankle pain. We report four patients with persistent posterolateral ankle pain despite prolonged nonoperative treatment. An os trigonal syndrome was diagnosed by a positive response to a fluoroscopically guided local anesthetic injection in the region of synchondrosis between the os trigonum and the posterior talus. RESULTS: All four patients underwent excision of the os trigonum with complete resolution of symptoms and return to full activity. CONCLUSIONS: Fluoroscopically controlled injection can help confirm the suspected diagnosis of an os trigonum syndrome and may have positive predictive value regarding the outcome of excisional surgery. PMID- 10847527 TI - Entrapment of the index flexor digitorum profundus tendon after fracture of both forearm bones in a child. AB - Entrapment of the index FDP tendon in a radius fracture callus occurred after fracture of both forearm bones in a 4-year-old boy. Surgical release of the FDP tendon, three months after fracture, resulted in normal index finger motion. This clinical problem can be avoided by a detailed physical examination of children with forearm fractures, verifying full passive range-of-motion of the hand after cast immobilization. Prompt supervised active range-of-motion should be done to prevent adhesions at the fracture site. PMID- 10847525 TI - Contested claims in carpal tunnel surgery: outcome study of worker's compensation factors. AB - We retrospectively analyzed the importance of factors relating to worker's compensation for 273 wrists in 211 consecutive patients who underwent primary carpal tunnel release. Patients were divided into three groups: non-work related, worker's compensation-uncontested, and worker's compensation-contested. Contested claims were those in which the worker's compensation carrier denied authorization for surgery, and in which such authorization was given following intervention by a plaintiff's attorney. RESULTS: there were no statistically significant differences in postoperative return of grip strength and in postoperative return to work intervals in comparing groups I and II. However, the contested worker's compensation patients were much less likely (and much slower) to return to light duty and to return to full duty work. Return of grip strength was slower and less complete in this group as well. Within worker's compensation, a contested claim portends a poorer prognosis. Uncontested worker's compensation claimants have nearly as good a prognosis as non-compensation patients. PMID- 10847528 TI - Subchondral metastasis: report of five cases. AB - Subchondral metastasis is a rare occurrence and poses a diagnostic dilemma as initial films may show a lytic lesion in the subchondral region often misinterpreted as being benign. We present five cases of subchondral metastasis as well as a review of the literature. In our cases, we present subchondral metastasis in the elbow, shoulder, and hip joints. All patients had pain over the affected joint and most presented with a lytic lesion in the subchondral bone. Three patients have died since presentation and two are doing well at last follow up visit. Subchondral metastasis is a rare entity, but it should be included in the differential of a lytic lesion in the subchondral bone. PMID- 10847529 TI - Glomus tumor of the finger tip and MRI appearance. PMID- 10847530 TI - Case report: paraarticular soft-tissue osteoma of the hip. AB - A case of paraarticular soft-tissue osteoma of the hip is presented. The patient is a 30-year-old white male with a two year history of progressive left hip pain. Plain film and cross-sectional imaging in conjunction with pathologic correlation are used to make the diagnosis. The lesion lacks the typical zoning pattern of myositis ossificans, shows no direct communication with native bone, and is extraarticular in location as opposed to synovial osteochondromatosis. Soft tissue osteomas most commonly occur around the knee, the foot, and the ankle. Soft tissue osteomas are rare tumors and this case is unusual in that it occurs around the hip. PMID- 10847531 TI - Anticalculus agents. AB - Most dentifrices and some mouthrinses now contain 1 or more active anticalculus agents to achieve so-called tartar control. The aim of this article is to review the research associated with the development of anticalculus agents and to evaluate in some detail, the efficacy of those agents which are currently available in dentifrice products. PMID- 10847532 TI - Diamond-coated sonic tips are more efficient for open debridement of molar furcations. A comparative manikin study. AB - BACKGROUND/AIMS: The aim of this study was to compare the efficacy of open debridement of class II and III furcations by a new diamond-coated sonic instrument tip (Sonicparo) with a conventional sonic insert (Sonicrecall) and hand instruments (Gracey curettes). METHOD: Open debridement was simulated using a manikin head and standardised plastic teeth. The efficacy of debridement was examined by 3 parameters: residual root surface marker, 'tooth' weight loss and time taken for debridement. The teeth used were 1st and 2nd upper and lower molars. Upper left and lower right molars were chosen to represent class II furcation involvements and upper right and lower left molars class III furcation involvements. Each tooth was debrided 4 times with each instrument for both class of furcations. RESULTS: The results of this study showed significantly less residual marker area comparing Sonicparo with Gracey curettes (mean difference 2.52 mm2, p<0.001). However, no significant differences in marker removal were observed between Sonicrecall inserts and the other instruments. Sonicparo inserts also took significantly less time to debride the furcation than Gracey curettes (mean difference 98.3 s, p<0.001) or Sonicrecall instruments (mean difference 74.3 s, p<0.001). However no significant differences were found between instruments concerning weight loss of the plastic teeth. CONCLUSIONS: It was concluded from this in-vitro study that the Sonicparo insert was a more efficient instrument for open molar furcation debridement than Sonicrecall and hand instruments (Gracey curettes) when considering both marker removal and debridement time. PMID- 10847533 TI - Community periodontal index of treatment needs and prevalence of periodontal conditions. AB - BACKGROUND/AIMS: In 1977, the World Health Organization (WHO) proposed a new index, the community periodontal index of treatment needs (CPITN) to evaluate the periodontal treatment needs of populations. The aim of this study is to compare different approaches of recording and presenting the CPITN. METHODS: A sample of 2110 subjects aged 35-44 years were examined between September 1994 and July 1995, throughout the province of Quebec, Canada. For each tooth (3rd molars excluded), the presence of bleeding and calculus, the level of epithelial attachment, and the depth of periodontal pockets were measured. Periodontal pocket depths were measured from the edge of the free gingiva, at 2 sites (mesiovestibular and vestibular), as well as all around the tooth. RESULTS: Only 8.5% of adults had at least one tooth with a 6 mm or deeper periodontal pocket when probing on 2 sites, whereas if probing is done all around the tooth, this percentage is 2.5x higher (21.4%). The partial recording of pocket depths (10 index teeth recommended by WHO, or 2 quadrants chosen at random) resulted in an underestimation of the prevalence of subjects with at least one tooth with a periodontal pocket (CPITN score 3 and 4). Among subjects with at least one tooth with a 6 mm or deeper periodontal pocket, 12% were not detected with the 10 index teeth recording, and 25% go undetected with the measure on 2 quadrants. Finally, using the % of subjects with periodontal pockets overestimates the prevalence of deep pockets compared with using sextants. Indeed, close to 30.0% of sextants have no treatment needs, whereas only 5.2% of subjects are in this category. Similarly, 7.7% of sextants have at least one tooth with a 6 mm or deeper periodontal pocket, yet there are 3x more subjects in this category (21.4%). PMID- 10847534 TI - Increased amounts of laminin in GCF from untreated patients with periodontitis. AB - BACKGROUND/AIMS: Our aim was to compare the levels of laminin and interleukin-8 (IL-8) in GCF from inflamed shallow (GP) and deep pockets (PP) in patients with periodontitis to these levels in GCF from inflamed shallow pockets (GG) in subjects with gingivitis alone. METHOD: Lactoferrin was used as a marker for the number of neutrophils in the sites sampled. The periodontitis group consisted of 13 subjects, having at least 6 sites with a pocket depth > or = 5 mm. The healthy control group consisted of 12 subjects with no clinical signs of periodontal destruction. GCF was collected with paper strips and the volume was measured immediately after sampling. Laminin, lactoferrin and IL-8 were measured using specific antibodies with an ELISA. RESULTS: The total amount of laminin was significantly higher in PP than in GG, but no significant difference was seen between GP and PP. The concentration and the total amounts of lactoferrin were similar in the three groups. The ratio between laminin and lactoferrin was higher in the samples from the patient, suggesting that neutrophils in patients are more activated and degrades more of the membrane per cell. The total amounts of IL-8 were very similar in the 3 groups, while the concentration also tended to be higher in the GP. CONCLUSIONS: Our study showed higher amounts of laminin in GCF from patients with periodontitis suggesting the presence of hyperactive neutrophils during the transmigration process through the endothelium/epithelium. PMID- 10847535 TI - Periodontal status by CPITN, and associated variables in an Israeli permanent force military population. AB - AIMS: The objectives were to determine periodontal treatment needs using the CPITN index, of Israeli permanent force military personnel. METHODS: The study population consisted of 1300 military personnel aged 25-44 years (mean age 33.8 +/- 5.4), who attended obligatory routine medical and dental examinations. Clinical examinations were conducted by 3 calibrated examiners, employing flat dental mirrors, the specially designed WHO periodontal probes (FDI probes) and following CPITN criteria. The frequency distributions were studied with regard to age groups, gender and education, as well as differences in the severity of the disease. Also, the mean number of sextants affected per person by age was assessed. RESULTS: Only 1.19% of the subjects demonstrated healthy periodontal tissue. Shallow pockets were similarly found among all age groups, and the number of persons with deep pockets increased with age. Deep pockets were found almost 3 x more among males (18.66%) in comparison with females (6.19%). Persons with higher education (> 12 years), had less deep pockets and bleeding than individuals with less than 12 years of education (p<0.05). CONCLUSIONS: Compared with data from other countries reported by the WHO, this periodontal status indicates a relatively high level of treatment needs. This survey provides a substantial contribution to the national oral health data bank concerning the adult working population in Israel. PMID- 10847536 TI - Healing following GTR treatment of bone defects distal to mandibular 2nd molars after surgical removal of impacted 3rd molars. AB - AIM: The purpose of this study was to study the healing, following guided tissue regeneration (GTR) treatment, of bone defects distal to mandibular 2nd molars (M2s) after surgical removal of impacted mesioangularly or horizontally inclined third molars (M3s) in patients > or = 25 years. METHOD: 20 patients with bilateral soft tissue impacted M3s were included in the split-mouth study. The 2 sites to be treated in each patient were randomised before the 1st operation as to which would undergo the test procedure and which would be the control site. After surgical removal of M3 at test sites, a resorbable polylactic acid (PLA) barrier was attached to M2 to cover the post-surgical bone defect. The flap was then replaced and sutured to cover the barrier. Control sites underwent the same procedure, as did the test sites, with the exception that no barrier was placed. The clinical examinations performed were oral hygiene pre- and 12 months postoperatively and probing pocket depth 12 months postoperatively. The alveolar bone level (ABL) at the distal surface of the M2, as determined from radiographs taken at suture removal and 12 months postsurgery, was chosen to be the primary response variable. RESULTS: Most bone defects showed healing up to 10%-20% of the tooth length at both test and control sites. 2 test and 2 control sites showed no improvement in the bone level. The mean values of bone healing registered in mm from the cemento-enamel junction (CEJ) were 2.6 +/- 2.19 SD and 3.0 +/- 2.20 SD for test and control sites, respectively. Different factors affecting the healing result are discussed. PMID- 10847537 TI - Healing following GTR treatment of intrabony defects distal to mandibular 2nd molars using resorbable and non-resorbable barriers. AB - AIMS: The objectives of the present, randomised clinical trial were (i) to evaluate the healing of periodontal intrabony defects at the distal aspect of mandibular 2nd molars using a resorbable polylactic acid (PLA) barrier and a non resorbable polytetrafluoroethylene (e-PTFE) barrier and (ii) to compare the therapeutic effect of the bioresorbable versus the non-resorbable barrier. METHOD: 19 patients with intrabony defects distal to mandibular 2nd molars > or = 4 mm (on radiographs) were included in the study. The defects all remained 5 years after surgical removal of impacted 3rd molars. Following flap elevation and defect debridement, the defects were randomly covered with, either a resorbable PLA or a non-resorbable e-PTFE barrier. Flaps were repositioned and sutured to completely cover the barriers. Treatment was evaluated clinically after 1 year by measurements of probing depth (PD), probing attachment level (PAL), and probing bone level (PBL) and radiographically by measurements of bone levels on computer digitised images of radiographs taken immediately before and 1 year postsurgery. RESULTS: Both treatments resulted in significant PD reduction, PAL gain, and bone fill. The total PD reduction was 5.3 +/- 1.9 mm for the PLA treated sites and 3.7 +/- 1.7 mm for the e-PTFE treated sites (p<0.05). The corresponding values for PAL gain were 4.7 +/- 0.7 mm and 3.6 +/- 1.7 mm (p<0.05) and for PBL gain 5.1 +/- 1.2 and 3.3 +/- 2.0 mm (p<0.05). Radiographic bone fill averaged 3.4 +/- 1.2 for the PLA and 2.0 +/- 1.6 mm for the e-PTFE barriers (p<0.05). Radiographic bone level measurements were significantly smaller than the corresponding clinical measurements, indicating that radiographs tend to underestimate bone fill. CONCLUSIONS: GTR treatment of deep intrabony defects distal to mandibular second molars using resorbable PLA barriers resulted in significant PD reduction, PAL gain and bone fill at least equivalent to the results obtained using non resorbable e-PTFE barriers. PMID- 10847538 TI - Relationships between radiographic alveolar bone height and probing attachment level: data from healthy post-menopausal women. AB - OBJECTIVE: To determine the cross-sectional and longitudinal relationships between radiographic alveolar bone height and probing attachment level in a population of healthy postmenopausal women. MATERIALS AND METHODS: The 81 patients in this report were part of a prospective estrogen replacement interventional study. All patients were in good oral health at entry and received annual oral prophylaxis as part of the study. Standard probing measurements were made with a pressure sensitive probe at 6 sites on each tooth. Vertical bitewing radiographs were digitized, and 6 linear measurements corresponding to probing site measurements were made from the cementoenamel junction to the alveolar crest. These procedures were performed at baseline and at annual intervals for three years. Data were analyzed both by site and by patient. RESULTS: Moderately strong correlations were found between cross-sectional measurements of probing attachment height and radiographic bone height. Correlations were stronger for patient data (r=0.44-0.61) than for site data (r=0.19-0.36). No relationships were found between longitudinal changes in alveolar bone height and attachment level in either the site data (r=-0.10-0.04). or the patient data (r=-0.005 0.10). CONCLUSION: Healthy patients may experience sporadic, temporary changes in attachment level or alveolar bone height which are resolved without one affecting the other. PMID- 10847539 TI - The effect of local delivery of PDGF-BB on attachment of human periodontal ligament fibroblasts to periodontitis-affected root surfaces--in vitro. AB - BACKGROUND/AIMS: The purpose of this study was to determine at what concentration does platelet-derived growth factor-BB (PDGF-BB) provide for optimal stimulation of human periodontal ligament fibroblasts (PDL) to adhere to periodontitis affected root surfaces. METHOD: 80 root dentine specimens were prepared from extracted periodontally diseased teeth obtained from patients ranging in age between 35 to 60 years. The root dentine specimens were associated with the subgingival area opposing the periodontal pocket for each extracted tooth. 10 healthy root dentine specimens were obtained from teeth extracted for orthodontic reasons and served as controls. The specimens were distributed into 9 groups (10 specimens in each). In group 1, PDL fibroblasts were cultured on the specimen surface of a diseased treated control. In group 2, PDL fibroblasts were cultured on the specimen surface of a healthy control. In groups 3 to 9, PDL fibroblasts were cultured on a pre-treated specimen surface with concentrations ranging from 5, 10, 20, 50, 100, 200 and 300 ng/ml PDGF-BB, respectively. After 24 h incubation, the media were removed, specimens were fixed, processed for SEM viewing and photographed at 750x. Fibroblast adherence was measured by counting number of cells within a standard test area and cell morphology was scored. RESULTS: Findings suggest dentine specimens pretreated with 5, 10 and 20 ng/ml PDGF-BB were not significantly different in number of adherent cells from the diseased treated control. However, at concentrations of 50, 100, 200 and 300 ng/ml, a highly significant increase in number of adherent fibroblasts was detected when compared to the diseased treated control. At these concentrations, the cell morphology was comparable to that of the healthy control. CONCLUSIONS: PDGF-BB in concentrations equal to or greater than 50 ng/ml demonstrates a significant stimulation of PDL cells adherence to periodontal diseased root surfaces. Since the higher concentrations resulted in similar effects as obtained by 50 ng/ml, it may therefore be considered that this concentration provides for optimal stimulation of human periodontal ligament fibroblasts (PDL) to adhere to periodontitis-affected root surfaces. PMID- 10847540 TI - Periodontal treatment of rapid progressive periodontitis in 2 siblings with Papillon-Lefevre syndrome: 15-year follow-up. AB - AIMS: This paper reports the treatment of the periodontal component of the Papillon-Lefevre syndrome in 2 siblings (case A, born 1974; case B, born 1976). METHOD: The initial treatment, in 1982, consisted of extraction of all primary teeth, scaling and rootplaning of the erupted permanent teeth and systemic antibiotic therapy. During 15 years, continuous and intensive periodontal treatment consisted of chlorhexidine 0.2% rinses, bi-weekly professional prophylaxis, scaling and rootplaning or surgery if indicated. Systemic antibiotics often accompanied mechanical therapy after bacteriological analysis. RESULTS: In case A, a favourable number of permanent teeth could be maintained, but in case B, all permanent teeth were lost in spite of the intensive treatment. Darkfield microscopy at different intervals revealed high numbers of spirochetes and motile rods in both siblings. Only in case A were they temporarily reduced to zero after scaling and rootplaning combined with metronidazole. Anaerobic cultering revealed high numbers of Actinobacillus actinomycetemcomitans (A.a) in both patients. In 1994, 2 years after combined amoxicillin/metronidazole therapy, no A.a could be detected in case A. In case B, A.a could still be detected and was found to be resistant to metronidazole. One year after extraction of all permanent teeth, could no A.a be detected in case B. CONCLUSION: Intensive periodontal treatment combined with antibiotic therapy was not able to prevent complete tooth loss in case B. In case A, the treatment was more effective, resulting in preserving a number of permanent teeth in a stable clinical situation. In these 2 cases, no attempt was made to create an edentulous period between the periodontally-diseased mixed dentition and the eruption of the remaining teeth, which may have contributed to treatment failure. PMID- 10847541 TI - Diffuse but unilateral gingival enlargement associated with von Recklinghausen neurofibromatosis: a case report. AB - BACKGROUND: An 8-year old girl was referred for diagnosis and treatment to the Department of Periodontology Oral Surgery of the University Hospital of Liege with an unusual clinical situation: a major, non-inflammatory, diffuse but unilateral enlargement of the interproximal, marginal and attached gingiva around all teeth of the right side of both the upper maxilla and mandible, whereas the alveolar process of the left side of upper and lower arches appeared strictly normal. METHOD: The clinical examination showed delayed eruption of some permanent teeth in the 1st and 4th quadrants. Except for its asymmetric occurrence, this gingival enlargement strongly resembled phenytoin-induced enlargement or gingival fibromatosis. This unilateral expression was evocative of a vascular or neurologic pathology. Several large "cafe-au-lait" spots were found disseminated on the body. Several selective surgical removals of thick gingival caps impairing the eruption of some permanent teeth were performed, and the removed tissues were histologically analyzed. RESULTS: Because of the presence of the large "cafe-au-lait" spots, a clinical diagnosis of Von Recklinghausen's disease was given and later confirmed several times by the histological analysis of the gingival biopsies. Now, 6 years later, this gingival enlargement due to the development of intra-gingival neurofibromas is stable and all permanent teeth have had a normal eruption, but alveolar bone growth has been partly impaired by the presence of the tumor. CONCLUSIONS: The present case of unilateral diffuse hyperplasia is a unique clinical expression of neurofibromatosis type I, a slowly evolving neurodermic dysplasia. PMID- 10847542 TI - Matrix metalloproteinase-8 levels and elastase activities in gingival crevicular fluid from chronic adult periodontitis patients. AB - AIM: To make an initial assessment of the periodontal diagnostic potential of immunoreactive matrix metalloproteinase-8 (MMP-8) in gingival crevicular fluid (GCF) by comparison with elastase activity which has previously been associated with disease severity and progression. METHODS: GCF was collected from molar and premolar sites of 16 chronic adult periodontitis patients before treatment and 13 of this group 2 weeks after scaling and root planing. Samples were analysed for MMP-8 by immunofluorometric assay and for elastase activity with a fluorogenic substrate. RESULTS: Mean patient clinical parameters and GCF enzyme totals both decreased significantly after treatment. Total MMP-8 levels and elastase activities generally correlated significantly with gingival and bleeding indices. For GCF concentrations, only MMP-8 showed a significant fall after treatment, and some significant correlations with clinical parameters. Amounts of the 2 enzymes correlated significantly with each other. CONCLUSIONS: Similarities between MMP-8 and elastase probably reflect the fact that both enzymes are associated mainly with neutrophils: MMP-8 levels may have fallen more after treatment because the assay, unlike that for elastase, would most likely not have detected much enzyme bound to alpha-macroglobulin. The immunoassay for MMP-8 is more specific and convenient than functional collagenase assays, and might be suitable for monitoring the periodontal condition. PMID- 10847543 TI - Effect of an essential oil-containing antiseptic mouthrinse on induction of platelet aggregation by oral bacteria in vitro. AB - BACKGROUND: With an increasing body of data suggesting an association between periodontitis and cardiovascular disease, studies have been conducted to elucidate potential mechanisms by which oral bacteria might exert systemic effects. 2 oral bacteria, Streptococcus sanguis and Porphyromonas gingivalis, have been shown to induce platelet aggregation in vitro. This study was conducted to determine the effect of treatment with an essential oil mouthrinse (Listerine Antiseptic) on the platelet-aggregating activity of these organisms. METHOD: Bacteria were grown under standard culture conditions. S. sanguis ATCC strain 10556 was exposed for 3 min to the essential oil mouthrinse at either full strength or a 1:1 dilution, while P. gingivalis FDC strain 381 was exposed to the essential oil mouthrinse at a 1:10 dilution. Positive control cells were treated with Hanks balanced salt solution (HBSS). Aggregation was measured using a recording platelet aggregometer. The assay of each organism in its respective mouthrinse dilution(s) or HBSS was repeated 5 times. RESULTS: In all cases, the HBSS-treated organisms induced platelet aggregation, with mean(+/-S.E.) lag times of 12.30 (+/-1.36) min and 11.36 (+/-0.58) min for P. gingivalis and S. sanguis, respectively. In contrast, treatment with the essential oil mouthrinse completely inhibited the platelet aggregating activity of P. gingivalis and of S. sanguis exposed to the 1:1 mouthrinse dilution in all assays; the aggregating activity of S. sanguis treated with full-strength mouthrinse was completely inhibited in 4 of 5 assays, and inhibited by 75% in the 5th, for a mean inhibition of 95 +/- 1.5%. CONCLUSION: This study provides additional evidence that the essential oil mouthrinse can interfere with bacterial cell surface-associated activities which may have clinical relevance. PMID- 10847544 TI - Histopathological features of flexural melanocytic nevi: a study of 40 cases. AB - Melanocytic nevi in certain locations such as the genital and acral sites may have atypical histologic features simulating melanoma. We studied the microscopic findings of 40 melanocytic nevi of flexural sites (axilla, umbilicus, inguinal creases, pubis, scrotum and perianal area) to verify if flexural nevi show distinctive features similar to melanocytic nevi of the genital skin. The patients were young (mean age 20 years), the lesions were mostly removed for cosmetic reasons and we are not aware of any deaths or complications related to the removed nevi. We found that 22 (55.5%) out of 40 flexural nevi had "a nested and dyshesive pattern" similar to the melanocytic nevi of genital skin. This pattern was characterized by the confluence of enlarged nests with variation in size, shape and position at the dermo-epidermal junction and by the diminished cohesion of melanocytes. Dermatopathologists should pay attention to the "nested and dyshesive pattern" of flexural nevi that may mimick histologic changes of melanoma. PMID- 10847545 TI - Epithelioid blue nevus: a rare variant of blue nevus not always associated with the Carney complex. AB - Epithelioid blue nevus is a rare variant of blue nevus that has been recently described in patients with Carney complex. Some of the patients with Carney complex have multiple epithelioid blue nevi and a familial history of similar lesions is often recorded. Epithelioid blue nevus consists of an intradermal melanocytic nevus composed of polygonal epithelioid cells laden with melanin. Neoplastic cells show no maturation at the base of the lesion and, in contrast with the usual stromal changes in blue nevi, epithelioid blue nevus exhibits no fibrosis of the dermis. We have studied three cases of epithelioid blue nevus in three patients with no evidence of Carney complex. The lesions were solitary and there was no family history of similar lesions. Therefore, epithelioid blue nevus is a distinctive variant of blue nevus that may also appear as a sporadic lesion and is not always associated with Carney complex. PMID- 10847546 TI - Long-term preservation of direct immunofluorescence staining in slides stored at room temperature. AB - To determine whether reproducible results from direct immunofluorescent tissue staining could be obtained after storing the slides at room temperature. We examined the original slides of 22 cases noted to be positive for direct immunofluorescence. Diagnoses include pemphigus, pemphigoid, lupus, dermatitis herpetiformis, lichen planus, and vasculitis. These specimens, initially evaluated during the period January 1997 to September 1998, were prepared with a standard immunofluorescence staining technique, and then a permanent aqueous mounting medium was added. All specimens were stored at room temperature in vertical slide holding trays. We focused on the presence and relative intensity of the immunofluorescence staining, as well as the final diagnosis. We then compared our readings to that of the original reports. Twenty of the 22 cases studied (91%) were read as having the same diagnosis as the initial immunopathology report. Seventeen of the 22 cases (77%) were found to have the identical or slightly less fluorescence intensity. The original reports in 3 of the cases did not comment on the original intensity of fluorescence. Thus, a comparison of fluorescence preservation could not be made. In 2 of the cases, the quality of tissue preservation was poor, and though fluorescent staining was noted, we were unable to render a diagnosis. Our results suggest that direct immunofluorescent studies, using a permanent aqueous mounting medium, can be stored over long periods of time at room temperature without significant degradation of staining. PMID- 10847547 TI - Detection of TCR-gamma gene rearrangements in early mycosis fungoides by non radioactive PCR-SSCP. AB - Early mycosis fungoides (MF) can mimic numerous benign inflammatory dermatoses on routine histological examination. In this study, a recently developed non radioactive polymerase chain reaction-single strand conformational polymorphism (PCR-SSCP) technique was used to assess T-cell clonality in paraffin-embedded skin biopsies clinically and pathologically suspicious for early MF. Non radioactive PCR-SSCP is a simple, sensitive, reproducible and rapid procedure requiring minimal instrumentation. DNA was extracted from 22 skin biopsies of 20 patients with suspected patch stage MF and 15 skin biopsies of inflammatory dermatoses. V gamma1-8, V gamma9, V gamma10, V gamma11 and J gamma1/J gamma2 consensus primers were used for T-cell receptor (TCR)-gamma gene rearrangement amplification. PCR products were analyzed by non-radioactive SSCP. Clonal TCR gamma gene rearrangements were detected in 17 of 22 (77%) suspected MF specimens. Clonal SSCP banding patterns were different among individual patients. In addition, identical banded patterns were demonstrated in serial skin biopsies from the same patient. No dominant T-cell clones were found in the inflammatory dermatoses studied. Therefore, non-radioactive PCR-SSCP is a useful molecular diagnostic tool for assessment of T-cell clonality in paraffin-embedded specimens suspicious for early MF. The SSCP imprint of PCR products is specific for each TCR-gamma gene rearrangement, and may be used to evaluate concurrent/recurrent disease in individual patients. PMID- 10847548 TI - Expression of the pro-apoptotic caspase 3/CPP32 in cutaneous basal and squamous cell carcinomas. AB - Apoptosis, a process of programmed cell death, plays an important role in normal and pathologic tissue homeostasis. The apoptotic cascade is triggered by caspases, a family of cystein proteases, among which caspase 3 (CPP32) seems to be the most directly related to apoptosis. Until now, few, if any, data exist on the role of CPP32 in skin tumors. We studied by immunohistochemistry, Western blot and reverse transcriptase-polymerase chain reaction (RT-PCR) the expression of CPP32 in specimens of normal human skin, basal and squamous cell carcinomas. CPP32 (both at the protein and mRNA level) was detected within epidermal and adnexal keratinocytes. CPP32 was also expressed in tumor masses of squamous cell carcinomas and more weakly in basal cell carcinomas; no correlation was found between CPP32 expression and depth of tumor invasion. CPP32 was occasionally expressed in tumor-infiltrating lymphocytes and peritumor endothelial cells. The expression of CPP32 in normal skin and in tumors arising from it suggests that this enzyme is involved in their homeostasis; its precise functional significance awaits further investigation. PMID- 10847550 TI - Lymphoma- and leukemia-associated cutaneous atypical CD30+ T-cell reactions. AB - Cutaneous CD30+ lymphoid infiltrates appear cytologically atypical and occasionally may be misinterpreted as recurrent disease when they occur in patients treated for other primary hematologic malignancies. We recently encountered two such cases and present our findings. One patient with B-cell lymphoma and another with myeloid leukemia developed cutaneous eruptions after chemotherapy displaying highly atypical perivascular lymphoid cells on histology that mimicked recurrent disease. In both cases, the lymphocytes were CD30+ T cells by immunohistochemistry. The skin lesions spontaneously resolved and have not recurred. Because one case was initially misinterpreted as recurrent leukemia, we conclude that close clinical correlation and immunophenotypic confirmation should be done for atypical cutaneous lymphoid infiltrates in patients with primary hematologic malignancies. We discuss the differential diagnosis of atypical CD30+ infiltrates in this setting, which include recurrent lymphoma or myeloid leukemia, primary cutaneous anaplastic large cell lymphoma (ALCL), lymphomatoid papulosis (LyP), carbamazepine-induced CD30+ pseudolymphoma, viral infection and an atypical eruption of lymphocyte recovery. PMID- 10847549 TI - Topoisomerase II-alpha expression in melanocytic nevi and malignant melanoma. AB - Malignant melanoma (MM) is considered to be a chemotherapy-refractory tumor. New anti-cancer drugs (e.g. etoposide) that target DNA topoisomerases (e.g. topoisomerase II-alpha (topo IIalpha)) show activity against a wide variety of solid tumors. In this study, we investigated the frequency and rate of labeling for topo IIalpha in 163 MMs (primary and metastatic) and 67 melanocytic nevi to determine whether topo IIalpha expression is elevated in MM. Primary MM exhibited significantly more frequent topo IIalpha expression compared to benign nevi (86% vs. 56%, p=0.0001). The rate of topo IIalpha labeling in dysplastic melanocytic nevi, radial growth phase MM, vertical growth phase MM and metastatic MM revealed significant differences amongst groups and a positive covariance with advancing stage (means: 0.3, 0.5, 5, and 8 '+' cells/hpf, respectively; r=0.3, all p < or = 0.02). Topo IIalpha labeling significantly correlated with increasing mitotic activity, depth of invasion and Clark's level, diminishing tumor infiltrating lymphocytes, and poor outcome (all p < or = 0.01) in primary MM. For metastatic MM, a minority (30%) exhibited marked elevation of topo IIalpha expression. These findings indicate topo IIalpha as a potential therapeutic target and marker for MM. Immunohistochemical analysis of disseminated MM may allow for correlation with clinical response and enable selection of candidates sensitive for specific chemotherapy. PMID- 10847551 TI - Giant cell lichenoid dermatitis within herpes zoster scars in a bone marrow recipient. AB - Cutaneous lesions arising in herpes zoster (HZ) scars are rare. We report a 34 year-old woman with acute lymphoblastic leukemia underwent allogenic bone marrow transplant (BMT). Ten days after the BMT, she developed clusters of vesicles over the right neck, scapula, shoulder and chest. She was treated with intravenous acyclovir and foscarnet. One month after the vesiculous episode of HZ she showed 5 mm to 2 cm clustered flat violaceous lichenoid papules and confluent plaques within the HZ scars. Histopathologic examination revealed a inflammatory infiltrate present in the papillary dermis with granulomatous aggregated formed by histiocytes, multinucleated giant cells and lymphocytes. She was treated with topic steroids with significant improvement. Pathologic findings are similar to those of an unusual lichenoid reaction named "giant cell lichenoid dermatitis". We present the first reported case of giant cell lichenoid dermatitis at the sites of HZ scars. PMID- 10847553 TI - Cutaneous metastases as initial manifestation of dedifferentiated chondrosarcoma of bone. An autopsy case with review of the literature. AB - Metastases of chondrosarcoma to the skin are uncommon. We report a case of dedifferentiated chondrosarcoma that manifested as cutaneous metastases and had an outcome of three weeks. A 69-year-old male presented with two cutaneous nodules, one in the chest and other in the inguinal area. The punch biopsy of the latter showed a poorly differentiated mesenchymal metastatic tumor. Shortly before death, an X-ray revealed a proximal epiphyseal lesion in the right humerus, radiographically interpreted as chondrosarcoma. The autopsy showed this lesion to be a dedifferentiated chondrosarcoma whose nonchondroid mesenchymal part was akin, histologically and immunohistochemically, to the cutaneous metastases. While ten previous reports of chondrosarcoma metastatic to the skin are known, we believe that this is the first case to report the cutaneous metastases of the dedifferentiated variety. Furthermore, skin metastasis preceding the diagnosis of chondrosarcoma has not been previously reported. The fact that one part of this kind of tumor can be highly undifferentiated or, else, differentiated along lines not usually reminiscent of bone tumors, can make the diagnosis of such cases extremely difficult. Most chondrosarcomas metastatic to the skin arise in bones of the extremities, including the hand. The most common type of tumor is conventional chondrosarcoma. These metastases can be either single or multiple with a slight predilection for the head and neck region. Most patients die in a mean time of 6 months after the appearance of cutaneous metastases. PMID- 10847552 TI - Multinucleate cell angiohistiocytoma: report of two cases with no evidence of human herpesvirus-8 infection. AB - Multinucleate cell angiohistiocytoma (MCA) is a vascular tumor of unknown pathogenesis. Possible misinterpretation of this disorder with Kaposi's sarcoma (KS), a human herpesvirus-8 (HHV-8)-associated tumor, prompted us to look for this virus in two women with MCA. None of the multiple skin specimens obtained from both our patients produced amplified HHV-8 DNA. Using a cell culture methodology similar to that used for KS, we established cell cultures from MCA lesions. While KS spindle cells are known to exhibit in vitro invasive properties and can be grown up to more than 20 passages, the MCA-derived cells were short lived and were not able to traverse basement membranes. Taken together, our data support the hypothesis that MCA is not a neoplasm but a benign vascular proliferation which is clearly distinguishable from KS. PMID- 10847554 TI - Chronic glycine treatment desensitizes the behavioral response to 1 aminocyclopropanecarboxylic acid (ACPC), a partial agonist at the strychnine insensitive glycine site of the NMDA receptor complex. AB - Chronic treatment with 1-aminocyclopropanecarboxylic acid (ACPC) but not with dizocilpine or imipramine produces desensitization to the behavioral response in ACPC challenge in the forced swim test (forced swim test). The mechanism by which ACPC produces this effect is unclear and may depend upon either its functional antagonist or its agonist properties at the NMDA receptor. We now report that chronic treatment with glycine or ACPC desensitizes the behavioral effect of challenge with ACPC in the forced swim test. The desensitization of the acute effects of ACPC cannot be explained by the presence of residual glycine because 24 h after the last of 14 daily glycine injections (i.e. the time of forced swim test) cortical and hippocampal glycine concentrations were unchanged. Likewise, the affinity of glycine to displace specific [3H]5,7-DCKA binding to glycine sites of the NMDA receptor complex was unchanged by chronic glycine administration. These results support the hypothesis that antidepressants produce adaptation of the NMDA receptor complex by mechanisms other than simply increasing synaptic glycine concentrations. Moreover, these results indicate that the behavioral adaptation in the forced swim test induced by chronic treatment with ACPC results from its agonist properties. PMID- 10847555 TI - Adult treatment with methamphetamine transiently decreases dentate granule cell proliferation in the gerbil hippocampus. AB - The objective of the present study was to examine whether acute treatment with the recreational drug methamphetamine influences adult granule cell proliferation in the dentate gyrus of the hippocampus. For that purpose, at the age of postnatal day 90 adult male gerbils (Meriones unguiculatus) received a single dose of either methamphetamine (25 mg/kg; i.p.) or saline. Proliferation of granule cells was identified by in-vivo labeling with 5-bromo-2'-desoxyuridine (BrdU) which was applied either simultaneously with methamphetamine or 36 h after administration of the drug. BrdU-labeled granule cell nuclei were identified in consecutive horizontal slices along the mid-septotemporal axis of the hippocampus and light-microscopically quantified 7 days after the BrdU-labeling. It was found that in both saline- and methamphetamine-treated animals there was a highly significant spatial septotemporal gradient in granule cell proliferation with numbers of BrdU-labeled cells gradually declining from the septal towards the temporal pole. The acute treatment with methamphetamine suppressed granule cell proliferation by about 28% and the septotemporal gradient of mitotic activity became significantly attenuated. It was further found that 36 h after the drug challenge granule cell proliferation rates had been restored almost to the control values along the whole septotemporal axis of the hippocampus. The present results are discussed with regard to (1) pharmacological regulation of neurogenesis in the hippocampus and (2) probable clues they may provide for both understanding the biological correlates of psychotic disorders and evolution of future concepts in neuropharmacological intervention. PMID- 10847556 TI - Inhibitory effect of a brain derived peptide preparation on the Ca++-dependent protease, calpain. AB - Overactivated calpain might be a key factor in destruction of cytoskeletal proteins involved in the pathophysiology of ischemia and disorders like Alzheimer's disease. Therapeutic effects imply the possible interference of Cerebrolysin (Ebewe Arzneimittel, Austria) with these molecular events. In this work several in vitro methods have been applied to investigate the interaction between Cerebrolysin and calpain [Enzyme Commission (EC) number: 3.4.22.17]. A conventional caseinolytic assay beside two flourimetric assays using a synthetic peptide substrate and a fluorescence labelled cytoskeletal protein [microtubule associated protein 2 labelled with 5-([4,6-dichlorotriazin-2-yl]amino) fluorescein (MAP2-DTAF)] respectively for a highly sensitive fluorimetric calpain activity assay were applied for kinetic analysis. The caseinolytic assay showed that the drug inhibits both mu- and m-calpain and to a significantly lower extent also trypsin [Enzyme Commission (EC) number: 3.4.21.1] and papain [Enzyme commission (EC) number: 3.4.22.6]. Dialysis experiments revealed Cerebrolysin mediated calpain inhibition to be reversible. Kinetic analysis exhibited a non competitive, or tight-binding competitive, mode of inhibition. This latter mode, substantiated by serial dilution experiments, and the likely existence of calpastatin in a brain derivative suggests the occurrence of calpastatin fragments or calpastatin-like fragments in Cerebrolysin. The clearly competitive inhibition of trypsin by the drug indicates distinct mechanisms and active components against different proteases. PMID- 10847558 TI - Serum levels of coenzyme Q10 in patients with Parkinson's disease. AB - We compared serum levels of coenzyme Q10 and the coenzyme Q10/cholesterol ratio in 33 patients with Parkinson's disease (PD) and 31 matched controls. The mean serum coenzyme Q10 levels did not differ significantly between the 2 study groups. Coenzyme Q10 levels were not correlated with age, age at onset, duration of the disease, scores of the Unified Parkinson Disease Rating Scale (UPDRS) or the Hoehn and Yahr staging in the PD group. The coenzyme Q10/cholesterol ratio had a significant correlation (although low) with duration of the disease (r = 0.46), total UPDRS score (r = -0.39), motor examination of the UPDRS (r = 0.45). These values were not influenced significantly by therapy with levodopa or dopamine agonists. The normality of serum coenzyme Q10 and coenzyme Q10/cholesterol ratio suggest that these values are not related with the risk for PD. PMID- 10847557 TI - Pramipexole attenuates the dopaminergic cell loss induced by intraventricular 6 hydroxydopamine. AB - The D3 preferring dopamine agonist pramipexole has been shown to attenuate the cell loss induced by levodopa in vitro. Pramipexole was herein evaluated in the 6 hydroxydopamine lesion model to determine its in vivo effect. Rats were treated with pramipexole or saline before and after an intracerebroventricular 6 hydroxydopamine injection. In the preliminary study, 6-hydroxydopamine produced a 68% reduction in striatal dopamine and a 62% loss in tyrosine hydroxylase immunoreactive (THir) cell counts in the substantia nigra. Pramipexole treated animals exhibited a 29% and a 27% reduction in striatal dopamine and THir cell counts, respectively. THir cell counts and striatal dopamine were significantly correlated. In the stereological study, 6-hydroxydopamine reduced THir cell counts by 47% in saline treated animals and 26% in pramipexole treated animals. These data demonstrate that pramipexole attenuates the biochemical and THir cell changes normally produced by 6-hydroxydopamine consistent with its neuroprotective actions in vitro. PMID- 10847559 TI - L-glutamate, L-arginine and L-citrulline levels in cerebrospinal fluid of Parkinson's disease, multiple system atrophy, and Alzheimer's disease patients. AB - Alterations in neuronal nitric oxide (NO) production may play a role in the pathophysiology of Parkinson's disease (PD) Alzheimer's disease (AD), and multiple system atrophy (MSA). The biosynthesis of NO is dependent on the availability of L-arginine, the substrate for NO-synthase (NOS), and on L glutamate, which stimulates NO synthesis via the NMDA receptor. In this process L citrulline is formed. We measured the levels of these amino acids in cerebrospinal fluid (CSF) of 108 PD patients, 12 AD patients, 15 MSA patients and 21 healthy subjects. A slight but statistically significant elevation of CSF L citrulline was found in MSA patients, while CSF L-glutamate was found to be significantly decreased in AD patients. We found no significant changes in L arginine levels. Although the relation between the CSF levels of these amino acids and neuronal NO production is still unclear, our findings suggest that AD is associated with a decrease in NO synthesis. PMID- 10847560 TI - Effects of fetal septal grafts on memory and learning performance with hippocampal acetylcholine and choline metabolism in fimbria transected rats. AB - Female Sprague-Dawley rats underwent aspirative lesion of the fimbria to produce septohippocampal disconnection. Two weeks after the lesion surgery, fetal septal grafts prepared from ventral forebrain of 13-15 days old fetuses of the same outbred strain were placed into the lesion cavity (grafted group). Three months after grafting, all rats were tested for spontaneous motor activity (SMA), step through passive avoidance (STPA) and in Morris' water maze (MWM). Six months after grafting, both basal and stimulated acetylcholine (ACh) and choline (Ch) release and their tissue levels were measured in ipsilateral hippocampal slices. Septohippocampal disconnection caused a significant impairment in Morris' water maze tasks, but did not alter spontaneous motor activity and step through passive avoidance. Fimbrial lesion, moreover, also declined both stimulated ACh release and tissue ACh levels in hippocampal slices. While lesion-induced change in Morris' water maze was ameliorated partially, declines in both stimulated ACh release and tissue ACh levels were raised to the control levels by fetal septal graft placed into the lesion cavity. These data show that grafted cholinergic neurons can work biochemically which may not result with a complete behavioral amelioration which is, in fact something more complex. PMID- 10847561 TI - An assessment of the antioxidant and the antiamyloidogenic properties of melatonin: implications for Alzheimer's disease. AB - This review summarizes recent advancements in our understanding of the potential role of the amyloid beta protein in Alzheimer's disease. It also discusses the significance of amyloid beta in initiating the generation of partially reduced oxygen species and points out their role in damaging essential macromolecules in the CNS which leads to neuronal dysfunction and loss. Recently acquired experimental data links these destructive oxidative processes with some neurodegenerative aspects of Alzheimer's disease. The experimental findings related to the free radical scavenging and antioxidative properties of melatonin are tabulated and its efficacy and the likely mechanisms involved in its ability to reduce neuronal damage mediated by oxygen-based reactive species in experimental models of Alzheimer's disease are summarized. Besides the direct scavenging properties and indirect antioxidant actions of melatonin, its ability to protect neurons probably also stems from its antiamyloidogenic properties. Melatonin is also unique because of the ease with which it passes through the blood-brain barrier. PMID- 10847562 TI - Serum levels of coenzyme Q10 in patients with Alzheimer's disease. AB - We compared serum levels of coenzyme Q10 and the coenzyme Q10/cholesterol ratio in 44 patients with Alzheimer's disease (AD), 17 patients with vascular dementia (VD), and 21 matched controls. The mean serum coenzyme Q10 and cholesterol levels and the coenzyme Q10/cholesterol ratio of patients with AD or VD did not differ significantly from those of controls. Coenzyme Q10 levels and coenzyme Q10/cholesterol ratio of AD or VD patients were not correlated with age, age at onset, duration of the disease or scores of the MiniMental State Examination. These results suggest that these values are not related with the risk for AD or VD. PMID- 10847563 TI - The anxiolytic effect of chronic inositol depends on the baseline level of anxiety. AB - Inositol, a precursor for membrane phosphoinositides involved in signal transduction, has been found to be clinically effective in a number of psychiatric disorders and to reverse behavioural effects of lithium. To gain insight into the mechanism of action of inositol, it is critical to establish its efficacy in animal models. Following the initial report by Cohen et al. (1997b) that inositol was anxiolytic in the elevated plus maze model of anxiety, the effect of chronic intraperitoneal and chronic dietary inositol administration in rats was tested in four experiments. There was a significant increase in closed arm and total arm entries following chronic injection of inositol, but no effect of inositol when it was given chronically in rat chow. Because the first 2 experiments suggested that the mode of drug administration affected the control levels of anxiety (open arm entries and time in open arms) in control groups, the effect of chronic dietary inositol was tested in rats that were exposed to a mild and a more severe form of stress. Chronic saline injections elevated anxiety in the plus maze, which was only marginally affected by chronic dietary inositol. Following 3 weeks administration of 5% dietary inositol rats were pre-exposed to a cat. There was a clear increase in number of entries into open arms, suggesting an anxiolytic effect of inositol. PMID- 10847564 TI - Compression friction and not proper fitting of the prosthesis. PMID- 10847565 TI - Syme's level amputation as ankle disarticulation. PMID- 10847566 TI - Creativity, collaboration, and cooperation. PMID- 10847567 TI - A preliminary investigation of pelvic obliquity patterns during gait in persons with transtibial and transfemoral amputation. AB - Differences in pelvic obliquity between small groups of persons with unilateral lower limb amputation and subjects without amputation were analyzed. Kinematic walking data were collected as six males with transtibial amputation and three males with transfemoral amputation walked over a range of speeds. The pelvic obliquity patterns and amplitudes from the groups with amputation were compared to normal data. Results showed that smaller peak-to-peak amplitudes of pelvic obliquity were associated with higher amputation levels. Pelvic drop during early prosthetic-limb stance tended to be smaller than during early sound-limb stance. Most of the subjects with amputation exhibited an obliquity pattern in which the hip on the prosthetic side was raised above the stance-side hip during prosthetic swing phase, indicative of a compensatory action known as hip-hiking. The subjects with transfemoral amputation exhibited this hip-hiking pattern during sound-limb swing phase as well. Results from this study suggest that further investigation is required to determine those limitations of current prosthetic technology that adversely affect pelvic obliquity in the gait of persons with amputation, and to determine if significant benefit can be realized by restoring a normal pattern of pelvic obliquity to the gait of persons with amputation. PMID- 10847568 TI - An analysis of the input-output properties of neuroprosthetic hand grasps. AB - We measured the input-output properties of the hand grasps of 14 individuals with tetraplegia at the C5/C6 level who had received an implanted upper limb neuroprosthesis. The data provide a quantitative description of grasp-opening and grasp-force control with neuroprosthetic hand grasp systems. Static properties were estimated by slowly ramping the command (input) from 0 to 100%. A hand-held sensor monitored the outputs: grasp force and grasp opening. Trials were performed at different wrist positions, with two different-sized objects being held, and with both grasp modes (lateral and palmar grasps). Larger forces were produced when grasping larger objects, and greater opening was achieved with the wrist in flexion. Although active grasp force increased with wrist extension, it was not significant statistically. Lateral grasp produced larger forces than the palmar grasp. The command range can be divided into a portion that controls grasp opening and a portion that controls grasp force. The portion controlling force increased with spacer size, but did not depend significantly on grasp mode or wrist position. The force-command relationships were more linear than the position-command relationships. Grasp opening decreased significantly over a one year period, while no significant change in grasp force was observed. These quantitative descriptions of neuroprosthetic hand grasps under varying conditions provide useful information about output capabilities that can be used to gauge the effectiveness of different control schemes and to design future control systems. PMID- 10847569 TI - Trends in lower limb amputation in the Veterans Health Administration, 1989-1998. AB - OBJECTIVE: To assess trends in lower limb amputation performed in Veterans Health Administration (VHA) facilities. METHODS: All lower limb amputations recorded in the Patient Treatment File for 1989-1998 were analyzed using the hospital discharge as the unit of analysis. Age-specific rates were calculated using the VHA user-population as the denominator. Frequency tables and linear, logistic, and Poisson regression were used respectively to assess trends in amputation numbers, reoperation rates, and age-specific amputation rates. RESULTS: Between 1989-1998, there were 60,324 discharges with amputation in VHA facilities. Over 99.9% of these were in men and constitute 10 percent of all US male amputations. The major indications were diabetes (62.9%) and peripheral vascular disease alone (23.6%). The age-specific rates of major amputation in the VHA are higher than US rates of major amputation. VHA rates of major and minor amputation declined an average of 5% each year, while the number of diabetes-associated amputations remained the same. CONCLUSION: The number and age-specific rates of amputations decreased over 10 years despite an increase in the number of veterans using VHA care. PMID- 10847570 TI - Assessment of the diabetic foot using spiral computed tomography imaging and plantar pressure measurements: a technical report. AB - Persons with diabetes mellitus (DM) and peripheral neuropathy are at high risk for skin breakdown due to unnoticed excessive pressures to the plantar foot during walking. We developed methods that combined spiral x-ray computed tomography (SXCT) imaging and plantar pressure analysis to quantify internal foot structure and external pressure during plantar loading. Methods were tested using a subject with DM who had a plantar ulcer, and a healthy control. SXCT measurements were within 2 mm of truth and SXCT plantar recordings were within 6.5% of walking trials. Hammer toe deformity (second toe), severe atrophy of the intrinsic muscles and less contact area during plantar loading, and a peak plantar pressure three times greater at the site of the ulcer were measured in the diabetic foot as compared with the healthy control. This preliminary investigation suggests that these methods are accurate for structural and pressure measurements of diabetic and healthy feet. PMID- 10847571 TI - Altered motor control and spasticity after spinal cord injury: subjective and objective assessment. AB - This study of measures of spasticity, or altered motor control, compares the clinically used Ashworth scale with a method based on surface electromyographic (sEMG) recordings called brain motor control assessment (BMCA) in a group of 97 subjects with spinal cord injury (SCI) and varying levels of motor dysfunction. In this paper, we describe how sEMG-derived scores relate to the severity of spasticity as judged clinically. When sEMG data from passive movements from the BMCA were analyzed by Ashworth category, we found that when the sEMG data were averaged for a limb, there was a significant difference between scores for those with Ashworth 0 vs. 2 and 3, and 1 vs. 2 and 3 (p<0.001), but not between 0 and 1. Analysis of the individual muscle scores improved the discrimination between Ashworth categories. Superiority of sEMG data over Ashworth category as an objective quantification of altered motor control ("spasticity") is argued. PMID- 10847572 TI - A practical EMG-based human-computer interface for users with motor disabilities. AB - In line with the mission of the Assistive Technology Act of 1998 (ATA), this study proposes an integrated assistive real-time system which "affirms that technology is a valuable tool that can be used to improve the lives of people with disabilities." An assistive technology device is defined by the ATA as "any item, piece of equipment, or product system, whether acquired commercially, modified, or customized, that is used to increase, maintain, or improve the functional capabilities of individuals with disabilities." The purpose of this study is to design and develop an alternate input device that can be used even by individuals with severe motor disabilities. This real-time system design utilizes electromyographic (EMG) biosignals from cranial muscles and electroencephalographic (EEG) biosignals from the cerebrum's occipital lobe, which are transformed into controls for two-dimensional (2-D) cursor movement, the left-click (Enter) command, and an ON/OFF switch for the cursor-control functions. This HCI system classifies biosignals into "mouse" functions by applying amplitude thresholds and performing power spectral density (PSD) estimations on discrete windows of data. Spectral power summations are aggregated over several frequency bands between 8 and 500 Hz and then compared to produce the correct classification. The result is an affordable DSP-based system that, when combined with an on-screen keyboard, enables the user to fully operate a computer without using any extremities. PMID- 10847573 TI - Compelled weightbearing in persons with hemiparesis following stroke: the effect of a lift insert and goal-directed balance exercise. AB - The hypotheses have been tested that 1) symmetry of weightbearing in persons who have sustained a stroke could be improved by the addition of a lift to the shoe on the non-paretic lower limb and 2) compelled weightbearing resulting from the addition of a lift in conjunction with targeted exercise helps to overcome the learned disuse of the paretic limb. Weightbearing on the paretic side was measured in eight persons with hemiparesis during quiet standing and in conditions of compelled weight shift. Compelled weight shifts were applied with special lifts to the shoe on the non-paretic limb of the subjects. An increase in symmetrical weightbearing was recorded in conditions of compelled weight shifts: 10-mm lift provided the best symmetry of bipedal standing. We suggest that improved symmetry of bipedal standing obtained with the lift of the non-paretic limb would help in overcoming the learned disuse of the affected limb. Pre- and post-test results of a person with hemiparesis who was wearing a shoe lift on the non-paretic limb during a 6-week physical therapy program showed statistically significant improvement of walking speed, stride length, and weightbearing. Such findings support the idea of using compelled weightbearing via lifting and targeted exercise during treatment. PMID- 10847574 TI - Effects of thoraco-lumbar electric sensory stimulation on knee extensor spasticity of persons who survived cerebrovascular accident (CVA). AB - Spasticity is mostly due to an excess of impulses to alpha motor neurons partly resulting from a change of interneuron activity. Low threshold afferent has been reported to change the activity in interneuron. The purpose of this study is to investigate the effects of surface spinal paravertebral stimulation on knee extensor spasticity. Ten survivors of stroke, with knee extensor spasticity, received electric stimulation for five 45-minute periods through surface electrodes applied to the skin in the twelfth thoracic and first lumbar areas. The electric stimulations had an amplitude modulated alternating current (AC), with a carrier frequency of 2500 Hz, modulated to "beats" frequency of 20 Hz. Stimulation amplitude was raised to elicit sensory stimulation. The pre- and post treatment evaluation included the modified Ashworth scale, active torque during controlled knee movements at various velocities, and electromyographic (EMG) activity during the torque measurements. Our results indicate that nine of ten subjects demonstrated a decrease in the modified Ashworth scale post-treatment. The EMG activity of the spastic quadriceps during active knee flexion was decreased post-treatment as compared with the value before treatment. The active torque value of knee flexion or extension at 30 degrees, 60 degrees, or 90 degrees/sec of angular velocity did not change significantly post-treatment. A trend of increasing spastic quadriceps EMG activity with respect to the angular velocity during an active knee flexion was established, with Ashworth scale considered. The level of EMG activity is higher when the Ashworth scale is higher. According to our results, the surface paravertebral sensory stimulation was effective in reducing quadriceps muscle spasticity of the subjects. Both the modified Ashworth scale and the EMG activity of spastic quadriceps during eccentric contraction are suggested as sensitive tools for measuring spasticity of persons who survived cerebrovascular accident (CVA). PMID- 10847575 TI - A technique for quantifying the response of seated individuals to dynamic perturbations. AB - A technique is presented for monitoring the seated postural stability and control of human subjects. Estimates are made of the locations of the subject's center of pressure (CP(S)) and projection of the center of mass (CM(NP)) from moment balance equations using measured force and acceleration data. The CP(S) and CM(NP) indices describe the stability of the subject, independent of the chair, even in the presence of perturbations. The measurement system was evaluated for both rigid objects and human subjects situated in a wheelchair undergoing displacement. Estimated CM(NP) was within +/-5 mm of the actual value for static loads. For human subjects, the average correlation coefficient between the estimated CM(NP) signal and that computed from video data was 0.90; however, transient overestimation of displacement was seen during subject acceleration. The technique could help to better assess seated stability in dynamic environments, such as those experienced by wheelchair users in motor vehicles. PMID- 10847576 TI - Wheelchair rider risk in motor vehicles: a technical note. AB - A better understanding of the risk involved in riding different sizes and types of motor vehicles is required to make informed decisions regarding a reasonable level of protection for wheelchair riders. Wheelchair rider accident information that can be used to estimate risk is quite limited. This paper reviewed the resources available, including the National Electronic Injury Surveillance System database. Motor vehicle accident data for the general public were analyzed in order to better characterize wheelchair rider risk. Using the National Safety Council annual transportation mode fatality rates and the (inverse) relationship of vehicle mass and occupant fatality rate, fatality rates for vehicles that transport wheelchair riders (minivans, vans, paratransit vans, and small and large buses) were estimated. Despite the large margins of error that must be assumed for accident data and the conclusions drawn from it, the available information suggests that 1) the majority of wheelchair rider injuries could be prevented by providing protection for abrupt vehicle maneuvers; 2) the type, size, and mass of the vehicle have a substantial effect on the fatality rate, although this effect decreases for heavier (<3,000 kg) vehicles; and 3) wheelchair riders who cannot properly use tiedown and occupant restraint systems or who are frail would face a lower risk of injury if transported in larger vehicles. PMID- 10847577 TI - Measuring the effectiveness of bioptic telescopes for persons with central vision loss. AB - PURPOSE: 1) To evaluate a vision rehabilitation program aimed at training persons with central vision loss to use a bioptic telescope for improving life skills, including driving and 2) to compare the outcomes of subjects who are given bioptic telescopes and training, with subjects who are prescribed telescopic lenses without training. METHODS: Twenty-five subjects ranging in age from 16 to 78 years were included in the study. Each subject was randomized to one of three groups: Group 1 received bioptic telescopes and training during the first approximately 3-month-long period of the approximately 6-month-long study; Group 2 received lenses and training during the second approximately 3-month-long period of the study; and Group 3 received the lenses for approximately 3 months without any training. An assessment battery consisting of clinical vision tests, functional tasks evaluated by an orientation and mobility specialist, driving skills evaluated by a kinesiotherapist specializing in driver's education, and psychophysical measures was administered to Groups 1 and 2 at baseline, and at approximately 3 and 6 months, and to Group 3 at baseline and at approximately 3 months. The tasks were categorized into 6 major functional categories: Recognition, Mobility, Peripheral Identification, Scanning, Tracking, and Visual Memory. Training consisted of 5 weeks of laboratory-based training focusing on skills within these 6 categories, and 8 weeks of on-road driving training. RESULTS: There was significant improvement in all task categories with use of the telescopes. There was improvement in all task groups with training, though a significant difference between the trained and untrained groups existed only in the Recognition, Peripheral Identification, and Scanning Categories, but not in Mobility, Tracking, or Visual Memory. When the tasks involving driving-related skills were analyzed separately, training also had a significant effect. CONCLUSION: There was significant improvement in visual skills with the use of a bioptic telescope. This improvement was greater with training in the use of the lenses in a number of visual skills categories including driving-related skills. PMID- 10847578 TI - The Timed Get-up-and-Go test revisited: measurement of the component tasks. AB - The "Timed Get-up-and-Go" (TGUG) test measures the overall time to complete a series of functionally important tasks. In the "Expanded Timed Get-up-and-Go" (ETGUG) test, times for the component tasks are measured using a multimemory stopwatch. Results from the ETGUG test were compared to those from the TGUG test on three groups of subjects: nonimpaired young, nonimpaired elderly, and elderly subjects at risk of falling. Significant differences were found between the two control groups and the at-risk group for all components of the test. Walking speed was the only measurement found to be significantly different between the young and elderly controls. The ETGUG test is a sensitive and objective assessment of function that requires minimal equipment, training, or expense. It better isolates functional deficits, thereby aiding the clinician in devising prevention strategies and guiding both treatment and further testing. PMID- 10847579 TI - Insulin and the beta3-adrenoceptor differentially regulate uncoupling protein-1 expression. AB - Cross-talk between insulin and the adrenergic system is important in the regulation of energy homeostasis. In cultured, differentiated mouse brown adipocytes, beta3-adrenergic stimulation induced a 4.5-fold increase in uncoupling protein-1 (UCP-1) expression, which was diminished by 25% in the presence of insulin. Beta3-adrenergic stimulation also activated mitogen activated protein (MAP) kinase by 3.5-fold and caused a decrease in basal phosphoinositide (PI) 3-kinase activity detected in p110gamma- and Gbeta-subunit immunoprecipitates in a time-dependent manner, whereas insulin stimulated p110alpha- and phosphotyrosine-associated PI 3-kinase activity. Inhibition of MAP kinase or PI 3-kinase potentiated the beta3-adrenergic effect on UCP-1 expression, both alone and in the presence of insulin. Thus, insulin inhibits beta3-adrenergic stimulation of UCP-1, and both MAP kinase and PI 3-kinase are negative regulatory elements in the beta3-adrenergic control of UCP-1 expression. Cross-talk between the adrenergic and insulin signaling systems and impaired regulation of UCP-1 might contribute to the development of a reduced energy balance, resulting in obesity and insulin resistance. PMID- 10847580 TI - Polypyrimidine tract-binding protein-associated splicing factor is a negative regulator of transcriptional activity of the porcine p450scc insulin-like growth factor response element. AB - The porcine P-450 cholesterol side-chain cleavage enzyme gene (P450scc) contains a 30-bp region [insulin-like growth factor response element (IGFRE)] that mediates insulin-like growth factor I (IGF-I)-stimulated gene expression and binds Sp1. In this study, we showed that polypyrimidine tract-binding protein (PTB)-associated splicing factor (PSF), an RNA-binding component of spliceosomes, binds to the IGFRE. Southwestern analysis with an IGFRE oligonucleotide showed that a protein (from Sp1-immunodepleted HeLa extract) fractionated on SDS-PAGE at 100 kDa. Microsequence analysis of 100-kDa band HeLa proteins detected PSF. DNA affinity chromatography, using an IGFRE mutant oligonucleotide that does not bind Sp1, isolated a protein that immunoreacted with PSF antibody. Deoxyribonuclease I (DNase I) footprint analysis showed recombinant PSF binds 5' of the Sp1-binding GC box of the IGFRE, and mutant oligonucleotides further delineated this region to a palindrome, CTGAGTC. Functional analysis of these mutants by transfection experiments in a cell line overexpressing the IGF-I receptor (NWTb3) found that an inability to bind PSF significantly increased the IGFRE transcriptional activity, while retaining responsiveness to IGF-I. Moreover, transfection of expression vectors for Sp1 and PSF in porcine granulosa cells found that Sp1 expression stimulated IGFRE transcriptional activity while PSF inhibited activity even with coexpression of Sp1. In conclusion, we identified PSF as an independent, inhibitory regulator of the transcriptional activity of the porcine P450scc IGFRE. PMID- 10847581 TI - A rapamycin-sensitive pathway down-regulates insulin signaling via phosphorylation and proteasomal degradation of insulin receptor substrate-1. AB - Insulin receptor substrate-1 (IRS-1) is a major substrate of the insulin receptor and acts as a docking protein for Src homology 2 domain containing signaling molecules that mediate many of the pleiotropic actions of insulin. Insulin stimulation elicits serine/threonine phosphorylation of IRS-1, which produces a mobility shift on SDS-PAGE, followed by degradation of IRS-1 after prolonged stimulation. We investigated the molecular mechanisms and the functional consequences of these phenomena in 3T3-L1 adipocytes. PI 3-kinase inhibitors or rapamycin, but not the MEK inhibitor, blocked both the insulin-induced electrophoretic mobility shift and degradation of IRS-1. Adenovirus-mediated expression of a membrane-targeted form of the p110 subunit of phosphatidylinositol (PI) 3-kinase (p110CAAX) induced a mobility shift and degradation of IRS-1, both of which were inhibited by rapamycin. Lactacystin, a specific proteasome inhibitor, inhibited insulin-induced degradation of IRS-1 without any effect on its electrophoretic mobility. Inhibition of the mobility shift did not significantly affect tyrosine phosphorylation of IRS-1 or downstream insulin signaling. In contrast, blockade of IRS-1 degradation resulted in sustained activation of Akt, p70 S6 kinase, and mitogen-activated protein (MAP) kinase during prolonged insulin treatment. These results indicate that insulin-induced serine/threonine phosphorylation and degradation of IRS-1 are mediated by a rapamycin-sensitive pathway, which is downstream of PI 3-kinase and independent of ras/MAP kinase. The pathway leads to degradation of IRS-1 by the proteasome, which plays a major role in down-regulation of certain insulin actions during prolonged stimulation. PMID- 10847582 TI - Functional trans-inactivation of insulin receptor kinase by growth-inhibitory angiotensin II AT2 receptor. AB - The present study demonstrates negative intracellular cross-talk between angiotensin II type 2 (AT2) and insulin receptors. AT2 receptor stimulation leads to inhibition of insulin-induced extracellular signal-regulated protein kinase (ERK2) activity and cell proliferation in transfected Chinese hamster ovary (CHO hAT2) cells. We show that AT2 receptor interferes at the initial step of insulin signaling cascade, by impairing tyrosine phosphorylation of the insulin receptor (IR) beta-chain. AT2-mediated inhibition of IR phosphorylation is insensitive to pertussis toxin and is also detected in neuroblastoma N1E-115 and pancreatic acinar AR42J cells that express endogenous receptors. We present evidence that AT2 receptor inhibits the autophosphorylating tyrosine kinase activity of IR, with no significant effect on insulin binding properties. AT2-mediated inactivation of IR does not mainly involve tyrosine dephosphorylation by vanadate sensitive tyrosine phosphatases nor serine/threonine phosphorylation by protein kinase C. As a consequence of IR inactivation, AT2 receptor inhibits tyrosine phosphorylation of insulin receptor substrate-1 (IRS-1) and signal-regulatory protein (SIRPalpha1) and prevents subsequent association of both IRS-1 and SIRPalpha1 with Src homology 2 (SH2)-containing tyrosine phosphatase SHP-2. Our results thus demonstrate functional trans-inactivation of IR kinase by G protein coupled AT2 receptor, illustrating a novel mode of negative communication between two families of membrane receptors. PMID- 10847583 TI - The critical role of Shc in insulin-like growth factor-I-mediated mitogenesis and differentiation in 3T3-L1 preadipocytes. AB - Insulin-like growth factor-I (IGF-I) stimulates mitogenesis in proliferating preadipocytes, but when cells reach confluence and become growth arrested, IGF-I stimulates differentiation into adipocytes. IGF-I induces signaling pathways that involve IGF-I receptor-mediated tyrosine phosphorylation of Shc and insulin receptor substrate 1 (IRS-1). Either of these adaptor proteins can lead to activation of the three-kinase cascade ending in activation of the extracellular signal-regulated kinase 1 and -2 (ERK-1 and -2) mitogen-activated protein kinases (MAPKs). Several lines of evidence suggest that activation of MAPK inhibits 3T3 L1 preadipocyte differentiation. We have shown that IGF-I stimulation of MAPK activity is lost as 3T3-L1 preadipocytes begin to differentiate. This change in MAPK signaling coincides with loss of IGF-I-mediated Shc, but not IRS-1, tyrosine phosphorylation. We hypothesized that down-regulation of MAPK via loss of proximal signaling through Shc is an early component in the IGF-I switch from mitogenesis to differentiation in 3T3-L1 preadipocytes. Treatment of subconfluent cells with the MEK inhibitor PD098059 inhibited both IGF-I-activation of MAPK as well as 3H-thymidine incorporation. PD098059, in the presence of differentiation inducing media, accelerated differentiation in subconfluent cells as measured by expression of adipocyte protein-2 (aP-2), peroxisome proliferator-activated receptor gamma (PPARgamma) and lipoprotein lipase (LPL). Transient transfection of subconfluent cells with Shc-Y317F, a dominant-negative mutant, attenuated IGF I-mediated MAPK activation, inhibited DNA synthesis, and accelerated expression of differentiation markers aP-2, PPARgamma, and LPL. We conclude that signaling through Shc to MAPK plays a critical role in mediating IGF-I-stimulated 3T3-L1 mitogenesis. Our results suggest that loss of the ability of IGF-I to activate Shc signaling to MAPK may be an early component of adipogenesis in 3T3-L1 cells. PMID- 10847584 TI - PDX-1 and cell-cell contact act in synergy to promote delta-cell development in a human pancreatic endocrine precursor cell line. AB - Cell lines from the fetal and adult pancreas that were developed by retroviral transfer of the SV40T and ras(val12) oncogenes lose insulin expression but retain extremely low levels of somatostatin and glucagon mRNA. In contrast to expanded populations of primary human islet cells, none of them express the homeodomain transcription factor PDX-1. When that factor was expressed in the cell lines by retroviral-mediated gene transfer, one of the cell lines, TRM-6, derived from human fetal islets, exhibited a 10- to 100-fold increase in somatostatin gene expression. This is the first report of induction of the endogenous somatostatin gene by PDX-1. Promotion of cell-cell contact by aggregation of TRM-6/PDX-1 into islet-like clusters produced a further 10- to 100-fold increase in somatostatin mRNA, to a level similar to that of freshly isolated islets, which resulted in production of somatostatin protein. Thus, we demonstrate here that signals induced by cell-cell contact act in synergy with PDX-1 to up-regulate the endogenous somatostatin promoter in an immortalized cell line from human fetal islets. This system provides a powerful model for studying human islet cell development and, particularly, the role of cell-cell contact in the differentiation process. PMID- 10847585 TI - Different subcellular localization and phosphoinositides binding of insulin receptor substrate protein pleckstrin homology domains. AB - Insulin evokes diverse biological effects through receptor-mediated tyrosine phosphorylation of the insulin receptor substrate (IRS) proteins. Here, we show that, in vitro, the IRS-1, -2 and -3 pleckstrin homology (PH) domains bind with different specificities to the 3-phosphorylated phosphoinositides. In fact, the IRS-1 PH domain binds preferentially to phosphatidylinositol 3,4,5-trisphosphate (PtdIns-3,4,5-P3), the IRS-2 PH domain to phosphatidylinositol 3,4-bisphosphate (PtdIns-3,4-P2), and the IRS-3 PH domain to phosphatidylinositol 3-phosphate. When expressed in NIH-IR fibroblasts and L6 myocytes, the IRS-1 and -2 PH domains tagged with green fluorescent protein (GFP) are localized exclusively in the cytoplasm. Stimulation with insulin causes a translocation of the GFP-IRS-1 and 2 PH domains to the plasma membrane within 3-5 min. This translocation is blocked by the phosphatidylinositol 3-kinase (PI 3-K) inhibitors, wortmannin and LY294002, suggesting that this event is PI 3-K dependent. Interestingly, platelet derived growth factor (PDGF) did not induce translocation of the IRS-1 and -2 PH domains to the plasma membrane, indicating the existence of specificity for insulin. In contrast, the GFP-IRS-3 PH domain is constitutively localized to the plasma membrane. These results reveal a differential regulation of the IRS PH domains and a novel positive feedback loop in which PI 3-K functions as both an upstream regulator and a downstream effector of IRS-1 and -2 signaling. PMID- 10847586 TI - Molecular cloning, expression, and characterization of rat homolog of human AP 2alpha that stimulates neuropeptide Y transcription activity in response to nerve growth factor. AB - Neuropeptide Y (NPY) plays an important role in the central regulation of neuronal activity, endocrine and sexual behavior, and food intake. Although transcription activity of the NPY gene in PC12 cells is regulated by a number of agents such as nerve growth factor (NGF), the mechanism responsible for the NGF elicited increase in the transcription of the NPY gene remains to be explored. In this study, we isolated and characterized a nuclear protein that is bound to NGF response elements (NGFRE) that lie between nucleotide -87 and -33 of the rat NPY promoter gene. This nuclear protein is identical to the rat homolog of human transcription factor AP-2alpha. We further demonstrated that rat AP-2a promotes efficient NPY transcription activity in response to NGF. Finally, we provide direct evidence that the mice lacking transcription factor AP-2alpha exhibit reduced expression of NPY mRNA compared with wild-type mice, further supporting the hypothesis that AP-2alpha is an important transcription factor in regulating NPY transcription activity. PMID- 10847587 TI - Hypoxia-inducible factor-1 (HIF-1) up-regulates adrenomedullin expression in human tumor cell lines during oxygen deprivation: a possible promotion mechanism of carcinogenesis. AB - Little is known about the molecular mechanisms that control adrenomedullin (AM) production in human cancers. We demonstrate here that the expression of AM mRNA in a variety of human tumor cell lines is highly induced in a time-dependent manner by reduced oxygen tension (1% O2) or exposure to hypoxia mimetics such as desferrioxamine mesylate (DFX) or CoCl2. This AM expression seems to be under hypoxia-inducible factor-1 (HIF-1) transcriptional regulation, since HIF-1alpha and HIF-1beta knockout mouse cell lines had an ablated or greatly reduced hypoxia AM mRNA induction. Similarly, inhibition or enhancement of HIF-1 activity in human tumor cells showed an analogous modulation of AM mRNA. Under hypoxic conditions, immunohistochemical analysis of tumor cell lines revealed elevated levels of AM and HIF-1alpha as compared with normoxia, and we also found an increase of immunoreactive AM in the conditioned medium of tumor cells analyzed by RIA. AM mRNA stabilization was shown to be partially responsible for the hypoxic up-regulated expression of AM. In addition, we have identified several putative hypoxia response elements (HREs) in the human AM gene, and reporter studies with selected HREs were capable of enhancing luciferase expression after exposure to DFX. Furthermore, transient coexpression of HIF-1alpha resulted in an augmented transactivation of the reporter gene after DFX treatment. Given that most solid human tumors have focal hypoxic areas and that AM functions as a mitogen, angiogenic factor, and apoptosis-survival factor, our findings implicate the HIF-1/AM link as a possible promotion mechanism of carcinogenesis. PMID- 10847588 TI - TASK (TWIK-related acid-sensitive K+ channel) is expressed in glomerulosa cells of rat adrenal cortex and inhibited by angiotensin II. AB - The present study was conducted to explore the possible contribution of a recently described leak K+ channel, TASK (TWIK-related acid-sensitive K+ channel), to the high resting K+ conductance of adrenal glomerulosa cells. Northern blot analysis showed the strongest TASK message in adrenal glomerulosa (capsular) tissue among the examined tissues including heart and brain. Single cell PCR demonstrated TASK expression in glomerulosa cells. In patch-clamp experiments performed on isolated glomerulosa cells the inward current at -100 mV in 30 mM [K+] (reflecting mainly potassium conductance) was pH sensitive (17+/-2% reduction when the pH changed from 7.4 to 6.7). In Xenopus oocytes injected with mRNA prepared from adrenal glomerulosa tissue the expressed K+ current at -100 mV was virtually insensitive to tetraethylammonium (3 mM) and 4-aminopyridine (3 mM). Ba2+ (300 microM) and Cs+ (3 mM) induced voltage-dependent block. Lidocaine (1 mM) and extracellular acidification from pH 7.5 to 6.7 inhibited the current (by 28% and 16%, respectively). This inhibitory profile is similar (although it is not identical) to that of TASK expressed by injecting its cRNA. In oocytes injected with adrenal glomerulosa mRNA, TASK antisense oligonucleotide reduced significantly the expression of K+ current at -100 mV, while the sense oligonucleotide failed to have inhibitory effect. Application of angiotensin II (10 nM) both in isolated glomerulosa cells and in oocytes injected with adrenal glomerulosa mRNA inhibited the K+ current at -100 mV. Similarly, in oocytes coexpressing TASK and ATla angiotensin II receptor, angiotensin II inhibited the TASK current. These data together indicate that TASK contributes to the generation of high resting potassium permeability of glomerulosa cells, and this background K+ channel may be a target of hormonal regulation. PMID- 10847589 TI - Novel role for the nuclear phosphoprotein SET in transcriptional activation of P450c17 and initiation of neurosteroidogenesis. AB - Neurosteroids are important endogenous regulators of gamma-aminobutryic acid (GABA(A)) and N-methyl-D-aspartate (NMDA) receptors and also influence neuronal morphology and function. Neurosteroids are produced in the brain using many of the same enzymes found in the adrenal and gonad. The crucial enzyme for the synthesis of DHEA (dehydroepiandrosterone) in the brain is cytochrome P450c17. The transcriptional strategy for the expression of P450c17 is clearly different in the brain from that in the adrenal or gonad. We previously characterized a novel transcriptional regulator from Leydig MA-10 cells, termed StF-IT-1, that binds at bases -447/-399 of the rat P450c17 promoter, along with the known transcription factors COUP-TF (chicken ovalbumin upstream promoter transcription factor), NGF-IB (nerve growth factor inducible protein B), and SF-1 (steroidogenic factor-1). We have now purified and sequenced this protein from immature porcine testes, identifying it as the nuclear phosphoprotein SET; a role for SET in transcription was not established previously. Binding of bacterially expressed human and rat SET to the DNA site at -418/-399 of the rat P450c17 gene transactivates P450c17 in neuronal and in testicular Leydig cells. We also found SET expressed in human NT2 neuronal precursor cells, implicating a role in neurosteroidogenesis. Immunocytochemistry and in situ hybridization in the mouse fetus show that the ontogeny and distribution of SET in the developing nervous system are consistent with SET being crucial for initiating P450c17 transcription. SET's developmental pattern of expression suggests it may participate in the early ontogenesis of the nervous, as well as the skeletal and hematopoietic, systems. These studies delineate an important new factor in the transcriptional regulation of P450c17 and consequently, in the production of DHEA and sex steroids. PMID- 10847590 TI - Characterization of transactivational property and coactivator mediation of rat mineralocorticoid receptor activation function-1 (AF-1). AB - The autonomous activation function-2 (AF-2) in the mineralocorticoid receptor (MR) E/F domain is known to play a major role in the ligand-induced transactivation function of MR; however, it remained unclear about the transactivation function of its A/B domain. We therefore tried to characterize the MR A/B domain as the AF-1 and further studied the actions of known coactivators for AF-2 in the E/F ligand-binding domain in the function of the MR A/B domain. Deletion analyses of rat and human MRs revealed that the A/B domains harbor a transactivation function acting as AF-1. The MR mutant (E959Q) with a point mutation in helix 12, which causes a complete loss of MR AF-2 activity, still retained ligand-induced transactivation function, indicating a significant role for AF-1 in the full activity of the ligand-induced MR function. Among the coactivators tested to potentiate the MR AF-2, TIF2 and p300 potentiated the MR AF-1 through two different core regions [amino acids (a.a.) 1-169, a.a. 451-603] and exhibited functional interactions with the MR A/B domain in the cultured cells. However, such interactions were undetectable in a yeast and in an in vitro glutathione-S-transferase pull-down assay, indicating that the functional interaction of TIF2 and p300 with the MR A/B domain to support the MR AF-1 activity require some unknown nuclear factor(s) or a proper modification of the A/B domain in the cells. PMID- 10847591 TI - The nuclear corepressors recognize distinct nuclear receptor complexes. AB - The thyroid hormone receptor (TR) and retinoic acid receptor (RAR) isoforms have the capacity to silence gene expression in the absence of their ligands on target response elements. This active repression is mediated by the ability of the corepressors, nuclear receptor corepressor (NCoR) and silencing mediator of retinoid and thyroid hormone receptors (SMRT), to recruit a complex containing histone deacetylase activity. Interestingly, NCoR and SMRT share significant differences in the their two nuclear receptor-interacting domains (IDs), suggesting that they may recruit receptors with different affinities. In addition, the role of the receptor complex bound to a response element has not been fully evaluated in its ability to recruit separate corepressors. We demonstrate in this report that the proximal ID in NCoR and SMRT, which share only 23% homology, allows preferential recognition of nuclear receptors, such that TR prefers to recruit NCoR, and RAR prefers to recruit SMRT, to DNA response elements. However, mutations in the TR found in the syndromes of resistance to thyroid hormone can change the corepressor recruited by changing the complex (homodimer or heterodimer) formed on the TRE. These results demonstrate that the corepressor complex recruited can be both nuclear receptor- and receptor complex specific. PMID- 10847592 TI - Activating protein-1, nuclear factor-kappaB, and serum response factor as novel target molecules of the cancer-amplified transcription coactivator ASC-2. AB - ASC-2 was recently discovered as a cancer-amplified transcription coactivator molecule of nuclear receptors, which interacts with multifunctional transcription integrators steroid receptor coactivator-1 (SRC-1) and CREB-binding protein (CBP)/p300. Herein, we report the identification of three mitogenic transcription factors as novel target molecules of ASC-2. First, the C-terminal transactivation domain of serum response factor (SRF) was identified among a series of ASC-2 interacting proteins from the yeast two-hybrid screening. Second, ASC-2 specifically interacted with the activating protein-1 (AP-1) components c-Jun and c-Fos as well as the nuclear factor-kappaB (NFkappaB) components p50 and p65, as demonstrated by the glutathione S-transferase pull-down assays as well as the yeast two-hybrid tests. In cotransfection of mammalian cells, ASC-2 potentiated transactivations by SRF, AP-1, and NFkappaB in a dose-dependent manner, either alone or in conjunction with SRC-1 and p300. In addition, ASC-2 efficiently relieved the previously described transrepression between nuclear receptors and either AP-1 or NFkappaB. Overall, these results suggest that the nuclear receptor coactivator ASC-2 also mediates transactivations by SRF, AP-1, and NFkappaB, which may contribute to the putative, ASC-2-mediated tumorigenesis. PMID- 10847593 TI - The C-terminal tail of the rat lutropin/choriogonadotropin (CG) receptor independently modulates human (h)CG-induced internalization of the cell surface receptor and the lysosomal targeting of the internalized hCG-receptor complex. AB - The analysis of 21 progressive truncations of the C-terminal tail of the rat LH/CG receptor (rLHR) revealed the presence of a region delineated by residues 628-649 that, when removed, enhanced the degradation of the internalized human (h)CG. The analysis of these truncations also revealed the presence of a region delineated by residues 624-631 that, when removed, enhanced the rate of internalization of hCG. Since there is little overlap between these two regions, we conclude that the structural features of the rLHR that mediate internalization and degradation of the internalized hormone are different. Detailed analyses of cells expressing a truncation at Y637 (designated rLHR-t637) showed that the enhanced degradation of hCG observed in the these cells is due to an increase in the rate of transfer of the internalized hCG-rLHR complex from the endosomes to the lysosomes rather than to the enhanced dissociation of the hCG-rLHR complex in the lysosomes. PMID- 10847594 TI - Early commitment of embryonic neocortical cells to develop area-specific thalamic connections. AB - In this study we examined the thalamic connectivity developed by grafts of embryonic (E16) parietal or occipital cortex placed homo- or heterotopically into the neocortex of newborn rats. We also examined the cytoarchitectonic organization developed by the grafts. Our findings indicate that E16 parietal cortex grafted into the parietal cortex of newborn recipients develops reciprocal connections with the host thalamic ventrobasal complex (VB) but does not establish connections with the host dorsal lateral geniculate nucleus (DLG). When implanted into the occipital cortex, these grafts are still able to establish connections with the VB. In contrast, E16 occipital cortex grafted into the parietal cortex establishes only a few connections with the VB. These grafts are, however, able to develop a substantial system of connections with the host DLG. At 16 days of embryonic age, graft cells are committed to establish thalamic connections appropriate to their tangential locus of origin. In addition, our results show that E16 parietal or occipital cortical cells do not possess the capacity to differentiate and maintain barrel organization even though they are grafted into the terminal field of developing VB axons. PMID- 10847595 TI - A new anatomical landmark for reliable identification of human area V5/MT: a quantitative analysis of sulcal patterning. AB - The location of human area V5 (or MT) has been correlated with the intersection of the ascending limb of the inferior temporal sulcus (ALITS) and the lateral occipital sulcus (LO). This study was undertaken to attempt a replication and quantification of these observations using functional magnetic resonance imaging. V5 was significantly activated in 19 hemispheres with alternating, low contrast, random checkerboard patterns. We confirmed the stereotaxic location of V5 and were able to describe a fairly consistent sulcal pattern in the parieto-temporo occipital cortex. V5 was usually (95%) buried within a sulcus, most commonly within the inferior temporal sulcus (ITS) (11%), the ascending limb of the ITS (ALITS) (53%) and the posterior continuation of the ITS (26%). The average distance from V5 of two identified anatomical landmarks of V5, the junctions of the LO and the ALITS, and the ITS and ALITS, were both 1 cm. However, the LO ALITS junction often had to be determined by interpolation (47%), and was not always present even with interpolation (21%). In contrast, the ITS-ALITS junction was always present and V5 was usually (90%) located in a sulcus intersecting with this junction, making it a more reliable landmark for localizing V5 with respect to gross morphological features on individual cortical surfaces. PMID- 10847596 TI - One-year age changes in MRI brain volumes in older adults. AB - Longitudinal studies indicate that declines in cognition and memory accelerate after age 70 years. The neuroanatomic and neurophysiologic underpinnings of cognitive change are unclear, as there is little information on longitudinal brain changes. We are conducting a longitudinal neuroimaging study of nondemented older participants in the Baltimore Longitudinal Study of Aging. This report focuses on age and sex differences in brain structure measured by magnetic resonance imaging during the first two annual evaluations. Cross-sectional results from 116 participants aged 59-85 years reveal significantly larger ventricular volumes and smaller gray and white matter volumes in older compared with younger participants and in men compared with women. Regional brain volumes show that the effects of age and sex are not uniform across brain regions. Age differences are greatest for the parietal region. Sex differences tend to be larger for frontal and temporal than parietal and occipital regions. Longitudinal analysis demonstrates an increase of 1526 mm(3) in ventricular volume over 1 year, but no detectable change in total or regional brain volumes. Definition of the pattern and rate of longitudinal brain changes will facilitate the detection of pathological brain changes, which may be predictors of dementia. PMID- 10847597 TI - Matching two imagined clocks: the functional anatomy of spatial analysis in the absence of visual stimulation. AB - Do spatial operations on mental images and those on visually presented material share the same neural substrate? We used the high spatial resolution of functional magnetic resonance imaging to determine whether areas in the parietal lobe that have been implicated in the spatial transformation of visual percepts are also activated during the generation and spatial analysis of imagined objects. Using a behaviourally controlled mental imagery paradigm, which did not involve any visual stimulation, we found robust activation in posterior parietal cortex in both hemispheres. We could thus identify the subset of spatial analysis related activity that is involved in spatial operations on mental images in the absence of external visual input. This result clarifies the nature of top-down processes in the dorsal stream of the human cerebral cortex and provides evidence for a specific convergence of the pathways of imagery and visual perception within the parietal lobes. PMID- 10847598 TI - Long-term depression in freely moving rats is dependent upon strain variation, induction protocol and behavioral state. AB - Hippocampal long-term depression (LTD) comprises a persistent decrease in synaptic transmission which is induced by repeated low-frequency stimulation (LFS). Although LTD has been widely demonstrated in the CA1 region in vitro, very few positive reports of LTD in vivo have occurred. In this study, the conditions under which homosynaptic LTD occurs in the CA1 region of freely moving rats was investigated. Three rat strains were studied: Wistar, Sprague-Dawley and Hooded Lister. Whereas Wistar and Sprague-Dawley rats expressed optimal LTD following 1 Hz LFS, Hooded Lister rats showed no LTD when tested in an LFS range of 1-10 Hz. Exposure to marked stress transiently enhanced LTD obtained in Wistar and Sprague Dawley rats, but did not facilitate LTD induction in the LTD-resistant strain. It was possible to induce long-term potentiation with high-frequency stimulation, although the profile of LTP was different in each strain. These data suggest that the expression of LTD varies according to the strain of rat used and is tightly dependent upon stimulation frequency. In addition the behavioral state of the animal may influence LTD expression. These data may explain, in part, the conflicting reports with regard to the inducibility of hippocampal LTD in vivo. PMID- 10847599 TI - Differential effects of discrete subarea-specific lesions of the rat medial prefrontal cortex on amphetamine- and cocaine-induced behavioural sensitization. AB - The medial prefrontal cortex (mPFC) of the rat is thought to be important for the initiation of behavioural sensitization. Since the mPFC is not a homogenous structure, we attempted to systematically examine the contribution of the different subareas - infralimbic (il), prelimbic (pl), anterior cingulate (cg) - of the mPFC to the induction of sensitization by selectively lesioning these areas or the whole mPFC with quinolinic acid (45 nmol in 0.5 microl). During an initial habituation session only il or whole mPFC lesions reduced spontaneous activity. Lesioned and sham-lesioned animals were then treated every other day with either saline, DL-amphetamine (3 mg/kg), or cocaine (20 mg/kg) for 2 weeks in their home cages and were then challenged with either DL-amphetamine (1.5 mg/kg) or cocaine (10 mg/kg) after 1 day and 2 weeks of withdrawal. None of the lesions affected the development of amphetamine-induced sensitization in any way, as assessed by several behavioural parameters including locomotion and sniffing. In contrast, cocaine-induced sensitization was significantly attenuated by pl and whole mPFC lesions, while il and cg lesions were without effect. These results show a double dissociation of the role of the mPFC in behavioural sensitization. The mPFC seems to be important only for cocaine- but not for amphetamine-induced sensitization, and only the pl area appears to be of relevance for cocaine induced sensitization. It is suggested that these differences are due to differences in the pharmacological interaction of cocaine and amphetamine with the mesocortical dopamine system, and to the particular anatomical connections of each of the mPFC subregions. PMID- 10847600 TI - The distribution of NADPH diaphorase and nitric oxide synthetase (NOS) in relation to the functional compartments of areas V1 and V2 of primate visual cortex. AB - The primary visual cortex (V1) of primates receives visual signals from cells in the koniocellular (K), magnocellular (M) and parvocellular (P) layers of the lateral geniculate nucleus (LGN). The functional role of the K pathway is unknown, but one proposal is that it modulates visual activity locally via release of nitric oxide (NO). One goal of this study was to examine the distribution of nitric oxide synthetase (NOS), the enzyme that produces NO, using immunocytochemistry for brain NOS (bNOS) or histochemistry for nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase activity in the V1 target cells of the K pathway and within the LGN itself. A second goal was to examine bNOS and NADPH diaphorase activity within proposed functional compartments in the second visual area (V2). We examined the LGN, V1 and V2 in squirrel monkeys, owl monkeys and bushbabies. In V1 and V2, we found that dense neuropil staining for NADPH diaphorase mirrored the pattern of high metabolic activity shown with cytochrome oxidase (CO) staining but did not necessarily mirror the pattern of immunolabeling seen with antibodies against NOS. The smooth stellate cells stained for NADPH diaphorase or bNOS were sparse and did not colocalize with LGN recipient zones in V1 or with the CO compartments in V2. LGN cells projecting to V1, including K, M and P cells, were negative for bNOS and NADPH diaphorase. Therefore, high levels of NOS are not limited to the K pathway. Instead, dense NOS activity is present in interneurons and within the neuropil of V1 and V2 that exhibit high metabolic demand. PMID- 10847601 TI - Human temporal lobe activation by speech and nonspeech sounds. AB - Functional organization of the lateral temporal cortex in humans is not well understood. We recorded blood oxygenation signals from the temporal lobes of normal volunteers using functional magnetic resonance imaging during stimulation with unstructured noise, frequency-modulated (FM) tones, reversed speech, pseudowords and words. For all conditions, subjects performed a material nonspecific detection response when a train of stimuli began or ceased. Dorsal areas surrounding Heschl's gyrus bilaterally, particularly the planum temporale and dorsolateral superior temporal gyrus, were more strongly activated by FM tones than by noise, suggesting a role in processing simple temporally encoded auditory information. Distinct from these dorsolateral areas, regions centered in the superior temporal sulcus bilaterally were more activated by speech stimuli than by FM tones. Identical results were obtained in this region using words, pseudowords and reversed speech, suggesting that the speech-tones activation difference is due to acoustic rather than linguistic factors. In contrast, previous comparisons between word and nonword speech sounds showed left lateralized activation differences in more ventral temporal and temporoparietal regions that are likely involved in processing lexical-semantic or syntactic information associated with words. The results indicate functional subdivision of the human lateral temporal cortex and provide a preliminary framework for understanding the cortical processing of speech sounds. PMID- 10847602 TI - Enhanced excitability of the human visual cortex induced by short-term light deprivation. AB - Long-term deprivation of visual input for several days or weeks leads to marked changes in the excitability and function of the occipital cortex. The time course of these changes is poorly understood. In this study, we addressed the question whether a short period of light deprivation (minutes to a few hours) can elicit such changes in humans. Noninvasive transcranial magnetic stimulation (TMS) of the human occipital cortex can evoke the perception of flashes or spots of light (phosphenes). To assess changes in visual cortex excitability following light deprivation, we measured the minimum intensity of stimulation required to elicit phosphenes (phosphene threshold) and the number of phosphenes elicited by different TMS stimulus intensities (stimulus-response curves). A reduced phosphene threshold was detected 45 min after the onset of light deprivation and persisted for the entire deprivation period (180 min). Following re-exposure to light, phosphene thresholds returned to predeprivation values over 120 min. Stimulus-response curves were significantly enhanced in association with this intervention. In a second experiment, we studied the effects of light deprivation on functional magnetic resonance imaging (fMRI) signals elicited by photic stimulation. fMRI results showed increased visual cortex activation after 60 min of light deprivation that persisted following 30 min of re-exposure to light. Our results demonstrated a substantial increase in visual cortex excitability. These changes may underlie behavioral gains reported in humans and animals associated with light deprivation. PMID- 10847603 TI - Determination of low-abundant metabolites in plant extracts by NAD(P)H fluorescence with a microtiter plate reader. AB - This article describes a method for the enzymatic detection of low-abundant metabolic intermediates in plant extracts via NAD(P)H fluorescence using a microtiter plate reader. The detection of changes in NAD(P)H fluorescence (excitation 340 nm, emission 465 nm) exhibits a high signal-to-noise ratio and is as sensitive (> or = 20 pmol per well) as absorbance measurements with dual wavelength photometers. Since up to 96 reactions can be initiated, monitored, and evaluated simultaneously, this method might be suitable for high-throughput screening programs on metabolite profiles. However, in contrast to absorbance measurements, fluorescence detection of NAD(P)H yields relative data, which can be impaired by the quench characteristics and the basic fluorescence of the extracts. Hence, extensive calibration is required to gain reproducible results. Calibration of the assay system was performed using leaf or root material (equivalent to 2-35 mg of fresh weight per well) extracted with perchloric acid, chloroform/water/methanol, or hot ethanol. Extraction with perchloric acid was found to be superior for metabolite quantification. Examples of the kinetics of individual metabolite determinations are presented and the contents of 3 phosphoglycerate, hexose phosphates, triose phosphates, pyruvate, and phosphoenolpyruvate in illuminated and darkened spinach leaves as well as leaf rosettes of Arabidopsis thaliana and leaf segments of the inducible crassulacean acid metabolism plant Mesembryanthemum crystallinum were measured via NAD(P)H fluorescence and, where possible, compared to reported data determined with dual wavelength photometers. PMID- 10847604 TI - Determination of urate crystal formation using flow cytometry and microarea X-ray diffractometry. AB - Gout is known to be induced by monosodium urate (MSU) crystals. The formation of MSU crystals is the first step of gouty inflammation. Detecting the early stage of crystallization accurately is considered to be important in understanding the mechanism of gouty arthritis. In this study, we employed flow cytometry (FCM) to detect small amounts of crystals produced in a supersaturated solution of uric acid. FCM was sensitive and crystals were determined at 2 h after the beginning of reaction. Gamma-globulin accelerated the formation rate time-dependently and dose-dependently. Low levels of lactic acid (less than 1.0 mg/ml) did not affect the formation rate but lactic acid of 2.0 mg/ml enhanced the formation of urate crystals. The crystals obtained with 2.0 mg/ml of lactic acid were analyzed with a microarea X-ray diffractometer and were shown to be a mixture of MSU and uric acid. FCM is a very useful method to determine the formation of crystals. Furthermore, analysis with a microarea X-ray diffractometer can provide detailed information about crystal composition. PMID- 10847605 TI - N-hydroxysuccinimidyl fluorescein-O-acetate as a fluorescent derivatizing reagent for catecholamines in liquid chromatography. AB - A new amine-reactive derivatizing reagent, N-hydroxysuccinimidyl fluorescein-O acetate (SIFA), was developed for catecholamine (CA) analysis in liquid chromatography. The reactivity of this reagent with the CAs norepinephrine (NE), epinephrine (E), and dopamine (DA) was investigated in detail. In aqueous methanol containing 32 mmol/L pH 9.0 H3BO3-Na2B4O7 buffer, SIFA reacted with NE, E, and DA under mild conditions. The derivatives were separated in 20 min on a C18 column with a mobile phase of methanol/water (38:62, v/v) containing 10 mmol/L pH 5.0 H3cit-Na2HPO4 buffer. At lambda(ex)/lambda(em) = 490/516 nm, the detection limits were 3.2, 12, and 56 fmol, respectively, with a signal-to-noise ratio of 3, which were comparable to those using 1,2-diphenylethylenediamine as the derivatizing reagent for CA analysis. Amino acids, aliphatic amines, and alcohols had no obvious interference with the determination. The proposed method has been applied to the determination of CAs in human urine, with recoveries of 95.3-103.9%. PMID- 10847606 TI - Atomic force microscopy of DNA molecules stretched by spin-coating technique. AB - We have developed an effective approach to stretching DNA molecules with the flow of fluid generated by spin coating. Well-stretched A DNA molecules were observed using atomic force microscopy. Substrate properties sensitively affected the stretching behavior of DNA. Our experimental findings revealed that a mica surface treated with crystal violet, a cationic dye molecule, is suitable to the spin-coating procedure for stretching DNA. Moreover, compared with relaxed DNA, we observed reduced height of the stretched DNA, which was attributed mainly to elongation force applied to the DNA molecules from the fluid flow and strong adhesion force between DNA and the substrate. This simple and effective method for preparing stretched DNA could be useful in physically mapping genomic DNA in a high throughput. PMID- 10847607 TI - Light-up probes: thiazole orange-conjugated peptide nucleic acid for detection of target nucleic acid in homogeneous solution. AB - We have constructed light-up probes for nucleic acid detection. The light-up probe is a peptide nucleic acid (PNA) oligonucleotide to which the asymmetric cyanine dye thiazole orange (TO) is tethered. It combines the excellent hybridization properties of PNA and the large fluorescence enhancement of TO upon binding to DNA. When the PNA hybridizes to target DNA, the dye binds and becomes fluorescent. Free probes have low fluorescence, which may increase almost 50-fold upon hybridization to complementary nucleic acid. This makes the light-up probes particularly suitable for homogeneous hybridization assays, where separation of the bound and free probe is not necessary. We find that the fluorescence enhancement upon hybridization varies among different probes, which is mainly due to variations in free probe fluorescence. For eight probes studied the fluorescence quantum yield at 25 degrees C in the unbound state ranged from 0.0015 to 0.08 and seemed to depend mainly on the PNA sequence. The binding of the light-up probes to target DNA is highly sequence specific and a single mismatch in a 10-mer target sequence was readily identified. PMID- 10847608 TI - A high-performance liquid chromatographic method to measure sphingosine 1 phosphate and related compounds from sphingosine kinase assays and other biological samples. AB - Sphingosine 1-phosphate is an intermediate of sphingosine catabolism as well as a potent signaling compound. Conditions were established for the extraction and analysis of sphingosine 1-phosphate and other sphingoid base 1-phosphates from in vitro sphingosine kinase assays and other biological samples. The sphingoid base 1-phosphates were extracted in high yield (85%) using small C-18 reverse-phase columns (LiChroprep RP-18). After the extracts were treated with 0.1 N KOH to remove glycerolipids, the sphingoid base 1-phosphates were converted to fluorescent o-phthalaldehyde derivatives that were separated by HPLC using C-18 columns with a mobile phase of methanol:10 mM potassium phosphate (pH 7.2):1 M tetrabutylammonium dihydrogen phosphate (in water) (83:16:1, v/v/v). The o phthalaldehyde derivative of sphingosine 1-phosphate was reasonably stable (t(1/2) > or = 18 h) when EDTA was present and could be detected in picomole amounts. The HPLC retention time of the sphingoid base 1-phosphates could be shifted by adjusting the mobile phase to pH 5.5, which is useful in separating overlapping compounds (such as sphingosine 1-phosphate and 4-D hydroxysphinganine) and in confirming the identity of sphingoid base 1-phosphates in biological samples. The extraction procedure and HPLC method facilitated assays of sphingosine kinase with different sphingoid bases as substrates and/or inhibitors and enabled the quantitation of sphingoid base 1-phosphates in human plasma, serum, and platelets as well as in strains of Saccharomyces cerevisae with mutations in sphingolipid metabolism. PMID- 10847609 TI - Observation of tetrahydrofolylpolyglutamic acid in bacteria cells by matrix assisted laser desorption/ionization mass spectrometry. AB - Tetrahydrofolylpolyglutamic acid in whole bacteria cells and cell lysates is analyzed by matrix-assisted laser desorption/ionization mass spectrometry. The speed, mass information, and tolerance to impurities of this technique make it ideal for monitoring the glutamation levels of folic acid in biological systems. Folylpolyglutamic acid is observed in a few strains of E. coli and two species of Staphylococcus bacteria. The effects of growth time, growth media, and the addition of methotrexate, a dihydrofolate reductase inhibitor, are also studied. PMID- 10847610 TI - Quantitation of inhibition of DNA methylation of the retinoic acid receptor beta gene by 5-Aza-2'-deoxycytidine in tumor cells using a single-nucleotide primer extension assay. AB - The expression of several cancer-related genes has been reported to be silenced by DNA methylation of their promoter region. 5-Aza-2'-deoxycytidine (5-AZA-CdR), a potent and specific inhibitor of DNA methylation, can reactivate the in vitro expression of these genes. In future clinical trials in tumor therapy with 5-AZA CdR a method to quantitate its inhibition of methylation of specific tumor suppressor genes would provide important data for the analysis of the therapeutic efficacy of this analogue. We have modified the methylation-sensitive single nucleotide primer extension assay reported by Gonzalgo and Jones (Nucleic Acids Res. 25, 2529-2531, 1997). Genomic DNA was treated with bisulfite and a fragment of the promoter region of the human retinoic acid receptor beta (RARbeta) gene, a tumor suppressor gene, was amplified using seminested PCR. Using two different primers we quantitated the inhibition of methylation produced by 5-AZA-CdR at two specific CpG sites in the RARbeta promoter in a human colon and a breast carcinoma cell line. The results obtained with the modified assay show a precise and reproducible quantitation of inhibition of DNA methylation produced by 5-AZA CdR in tumor cells. PMID- 10847611 TI - High-throughput screening of enzyme inhibitors: automatic determination of tight binding inhibition constants. AB - Determination of tight-binding inhibition constants by nonlinear least-squares regression requires sufficiently good initial estimates of the best-fit values. Normally an initial estimate of the inhibition constant must be provided by the investigator. This paper describes an automatic procedure for the estimation of tight-binding inhibition constants directly from dose-response data. Because the procedure does not require human intervention, it was incorporated into an algorithm for high-throughput screening of enzyme inhibitors. A suitable computer program is available electronically (http://www.biokin.com). Representative experimental data are shown for the inhibition of human mast-cell tryptase. PMID- 10847612 TI - Unique substrate specificities of two adjacent glutamine residues in EAQQIVM for transglutaminase: identification and characterization of the reaction products by electrospray ionization tandem mass spectrometry. AB - Reversed-phase HPLC (RP-HPLC) and electrospray ionization tandem mass spectrometry (ESI-MS/MS) were used to characterize the transglutaminase (TGase) catalyzed dual modification of a peptide (EAQQIVM, named FibN) with monodansylcadaverine (MDC). The synthesized FibN peptide, which was derived from the N-terminal sequence of fibronectin, was used as the substrate for a guinea pig liver TGase (G-TGase). The time course of incorporation of MDC into FibN, detected by RP-HPLC, indicated two separate fluorescent product peaks. ESI-MS analysis of the isolated fractions indicated that products represented MDC incorporated FibN molecules in molar ratios of 1:1 ((MDC)-FibN) and 2:1 ((MDC)2 FibN). A sequence analysis of MDC-FibN, using ESI-MS/MS, showed that the first modified residue in FibN was mainly Gln3. The kinetic analysis of MDC incorporation suggested that dual incorporation would occur by mainly one route. A one-dimensional 1H NMR comparison of MDC-FibN and unmodified FibN suggested that the first incorporation of MDC at Gln3 altered the substrate reactivity of the Gln4 residue in FibN for the G-TGase-catalyzed reaction. Thus, a detailed analysis of the peptide products using RP-HPLC and ESI-MS/MS should provide a powerful tool for exploring the mechanism of the substrate requirements of TGases. PMID- 10847614 TI - Bioluminescence detection of proteolytic bond cleavage by using recombinant aequorin. AB - Detection of proteolytic bond cleavage was achieved by taking advantage of the bioluminescence emission generated by the photoprotein aequorin. A genetically engineered HIV-1 protease substrate was coupled with a cysteine-free mutant of aequorin by employing the polymerase chain reaction to produce a fusion protein that incorporates an optimum natural protease cleavage site. The fusion protein was immobilized on a solid phase and employed as the substrate for the HIV-1 protease. Proteolytic bond cleavage was detected by a decrease in the bioluminescence generated by the aequorin fusion protein on the solid phase. A dose-response curve for HIV-1 protease was constructed by relating the decrease in bioluminescence signal with varying amounts of the protease. The system was also used to evaluate two competitive and one noncompetitive inhibitor of the HIV 1 protease. Among the advantages of this assay is that by using recombinant methods a complete bioluminescently labeled protease recognition site can be designed and produced. The assay yields very sensitive detection limits, which are inherent to bioluminescence-based methods. An application of this system may be in the high-throughput screening of biopharmaceutical drugs that are potential inhibitors of a target protease. PMID- 10847613 TI - A method for measuring disulfide reduction by cultured mammalian cells: relative contributions of glutathione-dependent and glutathione-independent mechanisms. AB - A method is described for measuring bioreduction of hydroxyethyl disulfide (HEDS) or alpha-lipoate by human A549 lung, MCF7 mammary, and DU145 prostate carcinomas as well as rodent tumor cells in vitro. Reduction of HEDS or alpha-lipoate was measured by removing aliquots of the glucose-containing media and measuring the reduced thiol with DTNB (Ellman's reagent). Addition of DTNB to cells followed by disulfide addition directly measures the formation of newly reduced thiol. A549 cells exhibit the highest capacity to reduce alpha-lipoate, while Q7 rat hepatoma cells show the highest rate of HEDS reduction. Millimolar quantities of reduced thiol are produced for both substrates. Oxidized dithiothreitol and cystamine were reduced to a lesser degree. DTNB, glutathione disulfide, and cystine were only marginally reduced by the cell cultures. Glucose-6-phosphate deficient CHO cells (E89) do not reduce alpha-lipoate and reduce HEDS at a much slower rate compared to wild-type CHO-K1 cells. Depletion of glutathione prevents the reduction of HEDS. The depletion of glutathione inhibited reduction of alpha lipoate by 25% and HEDS by 50% in A549 cells, while GSH depletion did not inhibit alpha-lipoate reduction in Q7 cells but completely blocked HEDS reduction. These data suggest that the relative participation of the thioltransferase (glutaredoxin) and thioredoxin systems in overall cellular disulfide reduction is cell line specific. The effects of various inhibitors of the thiol-disulfide oxidoreductase enzymes (1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU), arsenite, and phenylarsine oxide) support this conclusion. PMID- 10847615 TI - Fluorometric determination of deoxyribonuclease I activity with PicoGreen. AB - A rapid and sensitive assay for the detection of deoxyribonuclease I (DNase I) activity is described. This method is based on the ability of PicoGreen dye to enhance its fluorescence when bound to double-stranded DNA. In the standard assay, reaction mixtures containing the DNase I sample and 0.2 microg of the substrate DNA were prepared in a fluorescence microtiter plate and incubated at 37 degrees C. At the end of the reaction, the diluted PicoGreen reagent was added to each well and fluorescence intensity was measured with a fluorescence plate reader. By this assay, it was possible to determine precisely as little as 5 pg of DNase I within an hour. Moreover, using a small amount of the substrate DNA, the method was shown to be suitable for the sensitive detection of DNase I inhibitor activity. PMID- 10847616 TI - Determination of phosphorylation levels of tyrosine hydroxylase by electrospray mass spectrometry. AB - A novel approach has been developed to quantify the extent of phosphorylation of tyrosine hydroxylase (TH). The strategy consists of a chemical cleavage and characterization of the products using electrospray mass spectrometry (ESMS). The chemical cleavage involves selective hydrolysis of the aspartyl-peptide bond. Of the peptides formed, an 8-kDa NH2-terminus fragment is found to accurately duplicate the phosphorylation of TH using standard mixtures of TH-P/TH. The calibration yields a straight line with an R2 of 0.996, which is valid within the 10-90% range. The ESMS protocol has been used to determine the extent of phosphorylation of TH in the presence of CaM-PKII. The experimental conditions were designed to produce low levels of phosphorylation. Nevertheless, the ESMS analysis yielded single, double, and nonphosphorylation forms of TH. With respect to in vivo measurements, this ESMS protocol may be a generic procedure for determining the extent of phosphorylation of proteins. PMID- 10847618 TI - Amino acid analysis by capillary electrophoresis after phenylthiocarbamylation. AB - Capillary electrophoresis using SDS in phosphate buffer provides high resolution and short separation time for peptide and protein hydrolysate amino acids after derivatization with phenylisothiocyanate. The phenylthiocarbamyl derivatives are quantified in the picomole and femtomole range at signal-to-noise ratios better than 3:1 (for 50 fmol) and with a linearity correlation coefficient averaging 0.9938. The migration time and peak area variabilities were on average 1.1 and 2.7%, respectively. Complete separation of all the 18 amino acids normally found in polypeptide hydrolysates is achieved in less than 30 min using 75-microm capillaries while 50-microm capillaries require less than 15 min. Analysis of peptide and protein hydrolysates in the range 10-600 residues revealed excellent agreement with the known compositions at sensitivities better by large factors than the corresponding HPLC methodology (about 20-fold) and conventional ninhydrin-based analysis (about 1000-fold). PMID- 10847617 TI - Chromatographic and immunochemical approaches to the analysis of the HIV protease inhibitor saquinavir in plasma. AB - The development of the HIV protease inhibitor saquinavir (Ro 31-8959) required a range of analytical methods for its measurement in biological fluids. This paper describes the development of isocratic, reverse-phase HPLC/UV methods for the routine measurement of plasma levels of the drug together with a more sensitive radioimmunoassay. The performance of the two assays is compared with that of an HPLC/MS/MS method previously published and has been shown to be satisfactory, with coefficients of variation of calibration standards and quality control samples within the usual outside limits of +/- 15%. The HPLC/UV method can be routinely applied for concentrations down to 10-20 ng/ml and a lower limit of quantification of 1 ng/ml from 1 ml of human plasma is possible. The radioimmunoassay was developed for the specific measurement of saquinavir concentrations in human, HIV-positive plasma samples and has a lower limit of quantification of 0.5-1.0 ng/ml. Some preliminary findings suggested that it might not be specific in rat plasma and no attempts have been made to quantify any nonclinical samples with this technique. If still greater sensitivity is required, recourse can be made to the HPLC/MS/MS assay. PMID- 10847619 TI - A ganglioside-based assay for cholera toxin using an array biosensor. AB - A rapid assay for cholera toxin (CT) has been developed using a fluorescence based biosensor. This sensor was capable of analyzing six samples simultaneously for CT in 20 min with few manipulations required by the operator. The biochemical assays utilized a ganglioside-"capture" format: ganglioside GM1, utilized for capture of analyte, was immobilized in discrete locations on the surface of the optical waveguide. Binding of CT to immobilized GM1 was demonstrated with direct assays (using fluorescently labeled CT) and "sandwich" immunoassays (using fluorescently labeled tracer antibodies). Limits of detection for CT were 200 ng/ml in direct assays and 40 ng/ml and 1 microg/ml in sandwich-type assays performed using rabbit and goat tracer antibodies. Binding of CT to other glycolipid capture reagents was also observed. While significant CT binding was observed to loci patterned with GD1b, Gb3, and Gb4, CT did not bind significantly to immobilized GT1b at the concentrations tested. This is the first description of such a non-antibody-based recognition system in a multi-specific planar array sensor. PMID- 10847620 TI - Distinguishing the early-onset/persistent and adolescence-onset antisocial behavior types: from birth to 16 years. AB - Moffitt's theory regarding two types of adolescent antisocial behavior was investigated using a prospective, longitudinal study of normal and abnormal development in a primarily low socioeconomic status, ethnically diverse sample. Results supported the presence of an early-onset/persistent (EOP) group and an adolescence-onset (AO) group. Groups were most reliably and significantly distinguished by indices of socioemotional history within the first 3 years, but no significant differences were found on early measures of temperament or neuropsychological functioning. EOPs scored significantly lower than other groups on measures of neuropsychological functioning only during late childhood and adolescence, suggesting that the declines in verbal functioning that have been so reliably found in this and other samples of early-starting antisocial adolescents are progressive and consequent to adverse experience. In adolescence, AOs were significantly more likely to report high levels of internalizing symptoms and life stress, suggesting that AO antisocial behavior is not a benign phenomenon. Implications of these findings for etiologic theories of adolescent antisocial behavior are discussed. PMID- 10847621 TI - The development of attachment in new relationships: single subject analyses for 10 foster infants. AB - This paper presents single-subject analyses of newly developing attachment relationships in 10 foster infant-caregiver dyads. Using a diary methodology, at least 2 months of daily data were provided by foster parents on infants' attachment behaviors. Foster infant attachment was also assessed using the Strange Situation. Foster mother state of mind regarding attachment was measured using the Adult Attachment Interview. For eight infants, diary data revealed predominant patterns of attachment behavior emerging within 2 months of placement. In most cases, diary data predicted Strange Situation classifications. Both Strange Situation and diary data indicated that the three children placed in foster care before 12 months of age with foster parents having primary or secondary autonomous states of mind were classified as having secure attachments. The five children placed after 12 months of age showed predominantly insecure attachment behavior in the diary and were classified as insecure in the Strange Situation. Contingency analyses of behavioral sequences reported in the diary revealed that foster parents tended to complement their foster childrens' attachment behaviors. PMID- 10847622 TI - Severe maternal psychopathology and infant-mother attachment. AB - Eighty-two mother-infant dyads, comprising women with psychiatric disorder and individually matched controls, were followed up over the children's 1st year of life. The mothers with mental illness consisted of two subgroups: first, 25 severely mentally ill mothers who had been admitted to a psychiatric unit with their infants; and second, 16 mothers from a community sample meeting research diagnostic criteria for unipolar, nonpsychotic depression. With the exception of six dyads in the in-patient group, observations were made of the mother-infant interaction and the quality of the infant-mother attachment relationship at 12 months. The nature and course of the mothers' illness was also documented. Although few residual symptoms of maternal mental illness were detected at 1 year postpartum, interactional disturbances were evident among the case group dyads. A strong association was revealed between infant-mother attachment quality and maternal diagnosis; a manic episode of illness in the postpartum period was related to security in the attachment relationship, and psychotic or nonpsychotic depression was related to insecurity. Concurrent patterns of mother-infant interaction provided support for this finding. PMID- 10847623 TI - Conversational responsiveness in specific language impairment: evidence of disproportionate pragmatic difficulties in a subset of children. AB - Eighteen children with specific language impairment (SLI), from 6 to 8 years of age, were compared with 9 control children matched on age and nonverbal ability (CA controls) and with 9 younger control children of comparable language level (LA controls). Half of the SLI group were rated on a teacher checklist as having pragmatic difficulties: these were referred to as the pragmatic language impairment (PLI) group; the remainder were the typical (SLI-T) group. Children's responses to adult soliciting utterances were compared. All children usually responded to conversational solicitations, but children in the PLI group were more likely than control children to give no response, and they also made very little use of nonverbal responses, such as nodding. Nonverbal responding was closely related to the quality of children's responses. Children who failed to use nonverbal responses also had a relatively high level of pragmatically inappropriate responses that were not readily accounted for in terms of limited grammar or vocabulary. This study lends support to the notion that there is a subset of the language-impaired population who have broader communicative impairments, extending beyond basic difficulties in mastering language form, reflecting difficulty in responding to and expressing communicative intents. The analytic methods developed for this project have promise for the study of pragmatic difficulties in other clinical groups. PMID- 10847624 TI - School-aged children's vulnerability to depressive symptomatology: the role of attachment security, maternal depressive symptomatology, and economic risk. AB - The influences of three risk factors (insecure attachment, maternal depressive symptoms, and economic risk) on children's depressive symptomatology were examined. Subjects were 85 children from a range of income levels; all subjects were between 7 and 9 years of age. Children's depressive symptomatology was assessed with the Dimensions of Depression Profile for Children and Adolescents. Results indicated that children at high economic risk were more likely to exhibit depressive symptomatology than were children at low economic risk. Security of attachment was significantly related to children's depressive symptomatology. In addition, a significant relation was noted between children's depressive symptomatology and the depressive symptomatology of their mothers. Contrary to expectations, no significant relation was found between maternal depressive symptomatology and security of attachment. A multiple regression analysis revealed that security of attachment, maternal depressive symptomatology, and economic risk accounted for 47% of the variability in children's depression scores. Secure attachment served as a buffer; economic risk was associated with depressive symptoms only among insecurely attached children. PMID- 10847625 TI - Toward an interpersonal life-stress model of depression: the developmental context of stress generation. AB - The validity of a developmentally based life-stress model of depression was evaluated in 88 clinic-referred youngsters. The model focused on (a) the role of child-environment transactions, (b) the specificity of stress-psychopathology relations, and (c) the consideration of both episodic and chronic stress. Semistructured diagnostic and life-stress interviews were administered to youngsters and their parents. As predicted, in the total sample child depression was associated with interpersonal episodic and chronic stress, whereas externalizing disorder was associated with noninterpersonal episodic and chronic stress. However, the pattern of results differed somewhat in boys and girls. Youngsters with comorbid depression and externalizing disorder tended to experience the highest stress levels. Support was obtained for a stress generation model of depression, wherein children precipitate stressful events and circumstances. In fact, stress that was in part dependent on children's contribution distinguished best among diagnostic groups, whereas independent stress had little discriminative power. Results suggest that life-stress research may benefit from the application of transactional models of developmental psychopathology, which consider how children participate in the construction of stressful environments. PMID- 10847627 TI - Molecular regulation of the human inducible nitric oxide synthase (iNOS) gene. AB - In critically ill patients, the human inducible nitric oxide synthase (iNOS) gene is expressed in nearly every organ and has been shown to be associated with the refractory hypotension of septic and hemorrhagic shock. The molecular regulation of iNOS expression is complex and occurs at multiple sites in the gene expression pathway. Work in our laboratory has demonstrated that a combination of cytokines synergistically activate iNOS expression, which has resulted in the cloning of the first human iNOS gene from cytokine-stimulated hepatocytes. In addition, we have demonstrated that iNOS expression is transcriptionally regulated and that the functional promoter elements are located upstream of -4.7 kilobases (kb) within a unique enhancer region containing four functional nuclear factor kappa B elements. These results contrast markedly with the murine iNOS promoter, where only 1.0 kb of 5'-flanking sequence is required for lipopolysaccharide and cytokine responsiveness. Furthermore, numerous mechanisms have evolved to down regulate iNOS expression. By elucidating these mechanisms, therapeutic strategies to govern iNOS expression may be developed. PMID- 10847626 TI - Relational Psychotherapy Mothers' Group: a developmentally informed intervention for at-risk mothers. AB - The Relational Psychotherapy Mothers' Group (RPMG), a developmentally informed, supportive psychotherapy designed to serve heroin-addicted mothers with children up to 16 years of age, aims at addressing psychosocial vulnerabilities, and facilitating optimal parenting, among at-risk mothers. We present preliminary evidence on the efficacy of RPMG as an "add on" treatment in comparison with standard methadone counseling alone. At the end of the 24-week treatment period, mothers receiving RPMG plus standard methadone counseling demonstrated lower levels of risk for child maltreatment, greater involvement with their children, and more positive psychosocial adjustment than women who received methadone counseling alone. Children of RPMG participants also reflected fewer problems in multiple areas. At 6 months posttreatment, RPMG recipients continued to be at a relative advantage, although the magnitude of group differences was often attenuated. Notably, urinalyses indicated that RPMG mothers showed greater improvements in levels of opioid use over time than comparison mothers. PMID- 10847628 TI - Anticytokine therapies for acute inflammation and the systemic inflammatory response syndrome: IL-10 and ischemia/reperfusion injury as a new paradigm. AB - The results of recent anticytokine trials for sepsis syndrome have been disappointing. Several Phase II and Phase III clinical trials have shown a modest benefit in various subsets of patients; however, there has been no reported benefit in the primary endpoint of 28-day all-cause mortality. The failure of these trials is clearly multifactorial, and causes include the overall complexity of the inflammatory response, heterogeneity of the patient populations, absence of a hypercytokine response at the time of drug treatment, and the relatively short half-life of the administered drugs. The failure of anticytokine therapies may represent inadequate application of the treatment modality rather than any inherent weakness of the treatment itself. We have recently initiated a Phase I clinical trial examining the role of the anti-inflammatory cytokine IL-10 during surgical repair of a thoracoabdominal aortic aneurysm. This study may overcome some of the-design limitations of previous anticytokine trials in sepsis, and serve as a paradigm for future anticytokine therapy trials. Although the incidence of thoracoabdominal aortic aneurysms is relatively low, the patient population is homogeneous and the surgical injury associated with its repair reproducible. Additionally, postoperative mortality and morbidity rates are significant. Most importantly, the operative repair is associated with an obligatory visceral ischemia and reperfusion injury that appears to be associated with a proinflammatory cytokine response and postoperative organ dysfunction. IL 10 is a pleuripotent anti-inflammatory cytokine that both inhibits TNFalpha and IL-1 synthesis, and antagonizes their actions through upregulation of cytokine antagonists. Furthermore, IL-10 administration has been associated with only minimal adverse side effects during Phase I and Phase II trials. PMID- 10847629 TI - Interleukin-6 signal transduction in human intestinal epithelial cells. AB - The gut is an important source of inflammatory cytokines, but there is scant information on the mechanisms of cytokine action in gut epithelium. We hypothesized that in human Caco-2 cells, IL-6 acts directly through stimulation of Stat phosphorylation and that bacterial lipopolysaccharide (LPS) causes Stat activation indirectly because of its ability to cause the autocrine secretion and action of interleukin (IL)-6. Stat1, Stat5a, and Stat5b, but not Stat3, were detected in Caco-2 cells. DNA-binding activity corresponding to activated Stat5 was stimulated in a biphasic manner by IL-6, with a transient early phase, followed by sustained activation between 8 and 48 h. LPS also stimulated Stat5 like binding, but there was no early phase of activation. Functional tests of Stat5 activation showed that IL-6 stimulated Stat5-dependent reporter gene transcription but had no effect on Stat1-dependent transcription. LPS did not stimulate Stat-dependent transcription, nor did it alter the transcriptional response to IL-6. Tyrosine phosphorylation of both Stat5a and Stat5b was induced by IL-6. We infer from these data that IL-6 acts on intestinal epithelia through a Stat5-mediated transcriptional mechanism, whereas LPS does not induce gene expression through autocrine activation of enterocyte Stat signaling. These data provide a basis for testing the in vivo regulation of gut signaling and the interaction of gut reticuloendothelial cells with epithelial signal transduction. PMID- 10847630 TI - Induced nitric oxide inhibits IL-6-induced stat3 activation and type II acute phase mRNA expression. AB - Inducible nitric oxide synthase (iNOS) can be coexpressed with acute phase reactants in hepatocytes; however, it is unknown if NO can regulate the acute phase response. We tested the hypothesis that iNOS-derived nitric oxide (NO) attenuates the acute phase response by inhibiting IL-6-enhanced Stat3 DNA-binding activity and type II acute phase mRNA expression. iNOS was overexpressed in cultured rat hepatocytes via transduction with a replication defective adenovirus containing cDNA for human iNOS (AdiNOS), and Stat3 DNA-binding activity was determined by electrophoretic mobility shift assay (EMSA). EMSAs demonstrated that AdiNOS inhibits IL-6-induced Stat3 activation and that this inhibition is reversible in the presence of the NOS inhibitor N(G)-monomethyl-L-arginine (L NMA). The induction of beta-fibrinogen mRNA by IL-6, a Stat3 dependent process, is attenuated in AdiNOS-transduced cells and partially reversed by L-NMA. Thus, iNOS overexpression suppresses IL-6-induced Stat3 activation and type II acute phase mRNA expression in cultured hepatocytes. This suppression may represent a mechanism by which NO down-regulates the acute phase response. PMID- 10847631 TI - Inhibition of LPS-induced NFkappaB activation by a glucan ligand involves down regulation of IKKbeta kinase activity and altered phosphorylation and degradation of IkappaBalpha. AB - Growing evidence supports the role of transcription factor activation in the pathophysiology of inflammatory disorders, sepsis, ARDS, SIRS, and shock. Kinase mediated phosphorylation of IkappaBalpha is a crucial step in the NFkappaB activation pathway. We investigated IKBalpha phosphorylation in murine liver and lung extracts after cecal ligation and puncture (CLP) in the presence and absence of a glucan ligand. ICR mice were subjected to CLP. Unoperated and sham-operated mice served as the controls. Glucan phosphate (50 mg/kg) was administered 1 h before or 15 min after CLP. CLP increased hepatic and pulmonary levels of phospho IkappaBalpha by 48-192%. Pre- or post-treatment with glucan phosphate decreased (P < 0.05) tissue phospho-IkappaBalpha levels in CLP mice. Phospho-IkappaBalpha in the glucan-CLP group were not significantly different from the unoperated controls. To investigate mechanisms we examined IKKbeta kinase activity, IkappaBalpha phosphorylation and degradation, and NFkappaB activity in a murine macrophage cell line, J774a.1, treated with LPS (1 microg/mL) and/or glucan phosphate (1 microg/mL) for up to 120 min. The glucan ligand blunted LPS-induced IKKbeta kinase activity, phosphorylation and degradation of IkappaBalpha, and NFkappaB nuclear binding activity. The data indicate that one mechanism by which (1-->3)-beta-D-glucan may alter the response to endotoxin or polymicrobial sepsis involves modulation of IKK3 kinase activity with subsequent decreases in IkappaBalpha phosphorylation and NFkappaB activation. PMID- 10847632 TI - The Candida albicans INT1 gene facilitates cecal colonization in endotoxin treated mice. AB - Increased intestinal colonization with Candida albicans is believed to be a major predisposing factor to systemic candidiasis. Previous evidence has implicated the C. albicans INT1 gene in hyphal development, epithelial adherence, and mouse virulence. The effect of INT1 on mouse cecal colonization was measured using a parent strain (CAF2, INT1/INT1), an int1 deletion homozygote (CAG3, int1/int1), and a heterozygous reintegrant (CAG5, int1/int1 + INT1). Forty-eight hours after oral inoculation of 10(7) C. albicans into normal mice, only low numbers of each strain were recovered from the cecal flora. In mice pretreated with oral bacitracin/streptomycin, cecal colonization of each C. albicans strain was increased compared to the corresponding strain inoculated into untreated mice, with the CAF2 parent strain greater (P < 0.01) than the two mutant strains, and with the heterozygous and homozygous mutants not different from each other. In mice pretreated with parenteral lipopolysaccharide (LPS), in addition to oral antibiotics, numbers of cecal CAF2, CAG5, and CAG3 were increased (P < 0.01) compared to the corresponding strain inoculated into mice treated with antibiotics alone. In LPS-treated mice, numbers of cecal C. albicans CAF2 (INT1/INT1) were greater (P < 0.05) than C. albicans CAG3 (int1/int1). Thus, parenteral LPS had an additive effect on C. albicans cecal colonization in antibiotic-treated mice, and the presence of two functional copies of the INT1 gene appeared to facilitate colonization in both antibiotic-treated mice and in mice treated with antibiotics plus parenteral endotoxin. PMID- 10847633 TI - Effects of MCI-154, a calcium sensitizer, on cardiac dysfunction in endotoxic shock in rabbits. AB - This study was designed to observe the effects of MCI-154, a calcium sensitizer, on cardiac dysfunction after endotoxic shock in rabbits. Ten hours after the rabbits were given injection of 1.0 mg/kg endotoxin (Escherichia coli, O111:B4) via marginal ear veins, 0.1 mg/kg MCI-154 was injected intravenously and then 50 mL/kg normal saline (NS) + 0.1 mg/kg MCI-154 was infused continuously at a rate of 0.7 mL/min. During this process, the parameters of cardiac function were measured. It was found that 10 h after the endotoxin injection, heart rate (HR) was increased significantly while the mean arterial blood pressure (MAP), left ventricular systolic pressure (LVSP), isovolumetric pressure (IP), myocardial contractility (MC), and the area of p-dp/dt(max) vector loop (Lo) all were markedly decreased. Treatment with 50 mL/kg NS alone had slight effects on these parameters. On the contrary, LVSP, IP, MC, and Lo all were increased significantly while HR was not obviously changed and left ventricular end diastolic pressure (LVEDP) was reduced remarkably following MCI-154 administration in endotoxic shock rabbits. The parameters of myocardial contractility were improved nearly to the values in sham shock group and were markedly higher than that in NS alone-treated group. It can be concluded that MCI 154 can exert significant therapeutic effects on cardiac dysfunction after endotoxic shock, for it improves cardiac function, dilates peripheral blood vessels, and slightly affects HR. PMID- 10847634 TI - Chelerythrine chloride induces rapid polymorphonuclear leukocyte apoptosis through activation of caspase-3. AB - Polymorphonuclear leukocytes (PMN) play a primary role in the initiation and propagation of inflammatory responses. PMN apoptosis is a major mechanism associated with the resolution of inflammatory reactions. Understanding mechanisms associated with PMN apoptosis will be of critical value in the development of novel pharmacological treatment strategies for local and/or systemic inflammatory disorders. The present study demonstrates that chelerythrine chloride induces human PMN to undergo rapid and synchronous progression into the apoptotic process via a PKC-independent mechanism. The appearance of the morphological features of apoptosis in chelerythrine-treated PMN is preceded by a significant upregulation in caspase-3 activity. GM-CSF (a cytokine that protects PMN in several models of PMN apoptosis) does not protect PMN from chelerythrine chloride-induced apoptosis. PMID- 10847635 TI - Effects of prolonged partial liquid ventilation, high frequency ventilation and conventional ventilation on gas exchange and lung pathology in newborn surfactant depleted piglets. AB - Partial liquid ventilation (PLV) improves oxygenation in various animal models of respiratory insufficiency. The aim of this study was to compare the effects of conventional ventilation (CV), high frequency oscillatory ventilation (HFOV), and PLV combined with CV or HFOV on gas exchange and histopathology. Thirty anaesthetised newborn piglets (mean weight 1.94 kg, age 1-3 days) were randomized in five groups of six animals: CV, CV + surfactant (S), HFOV+S, PLV/CV, and PLV/HFOV. Thirty min after lung injury had been induced with repeated saline lavage, specific ventilatory treatment was initiated. Three animals of the CV group died within the 24 h study period, whereas none died in any of the other groups. The oxygenation index (OI) and the PaO2/FIO2 ratio improved significantly within 30 min in all groups, but not in the CV group. After 24 h all oxygenation parameters were better in the PLV groups than in CV or CV+S (P < 0.05). No differences in gas exchange were noted between HFOV+S and PLV/CV. The combination of PLV with HFOV led to an increased PaO2/FIO2 ratio when compared with PLV/CV and with HFOV+S (P < 0.05). All PLV treated animals had significantly less lung injury in the upper and lower lobes compared with gas-ventilated animals by histologic semi-quantitative lung injury score (P < 0.01) and in the lower lobes by morphometry (P < 0.001). In conclusion, HFOV+S and PLV either with CV or HFOV are effective techniques to provide adequate gas exchange in S-deficient lungs compared with CV with and without S. However, lung injury was significantly improved in both PLV treated groups compared with HFOV+S and the CV groups. PMID- 10847636 TI - The cardiovascular hemodynamics and leukotriene kinetics during prostacyclin and anti-prostacyclin antibody infusions in septic shock. AB - This study evaluated whether or not prostacyclin (PGI2) was necessary or sufficient by itself in a pathophysiologic concentration to mediate the cardiovascular dysfunction of septic shock. Anesthetized adult swine received anesthesia only (ANESTHESIA CONTROL, n = 6); graded Aeromonas hydrophila, 10(10)/mL, infusion at 0.2 mL/kg/h that increased to 4.0 mL/kg/h over 3 h (SEPTIC SHOCK CONTROL, n = 6); pathophysiologic prostacyclin infusion to match septic shock control plasma levels without bacteremia (PGI2 INFUSION, n = 6), or graded Aeromonas hydrophila plus anti-prostacyclin antibody infusion (ANTI-PGI2-Ab INFUSION, n = 5). This graded porcine bacteremia model was 100% lethal after 4 h. Cardiovascular hemodynamics, arterial blood gases, and plasma levels of arachidonate metabolites were measured at baseline and hourly over a 4-h period. The results showed that PGI2 was not a necessary mediator of impaired cardiovascular hemodynamics in graded bacteremia, as anti-PGI2 antibody infusion did not improve the cardiac index, systemic vascular resistance, or peripheral oxygen balance in septic animals. Also, PGI2 was not sufficient alone to cause the cardiovascular dysfunction of sepsis, as pathophysiologic infusion of PGI2 did not reproduce such changes in normal animals. PGI2 blockade during bacteremia significantly increased LTC4D4E4, and LTB4 whereas PGI2 infusion suppressed LTC4D4E4 concentration, suggesting that endogenous PGI2 may blunt leukotriene release during septic shock. These results indicate a complex dynamic equilibrium among prostacyclin and leukotrienes in septic shock. PMID- 10847637 TI - High doses of intravenous immunoglobulin G enhance Kupffer cell phagocytic function during the late phase of sepsis and endotoxemia in rats. AB - The effect of intravenous immunoglobuln G (ivIG) on the hepatic microvascular inflammatory response during the late phase of sepsis and endotoxemia in rats was studied by using in vivo microscopy. One hour after administration of a clinically relevant dose of ivIG (0.5 g/kg body weight, Sandoglobulin), rats were subjected to polymicrobial sepsis induced by cecal ligation and puncture (CLP) or were injected intravenously with lipopolysaccharide (LPS, 0.1 mg/kg body weight). Twenty-four hours after CLP or LPS, the number of leukocytes adhering to the sinusoidal wall was increased 11.0-fold in CLP-treated animals and 5.6-fold in LPS-treated animals, respectively, compared with the controls. Concomitantly, the numbers of swollen sinusoidal endothelial cells were increased 4.2-fold and 3.2 fold. The number of perfused sinusoids was decreased by 35% and by 24%. These responses were minimized by pretreatment with high doses of ivIG. Kupffer cell phagocytic activity in the periportal sinusoids in CLP-treated animals was decreased by 41%, whereas that in the centrilobular sinusoids in LPS-treated animals was increased by 72%. IvIG significantly elevated this activity in both CLP- and LPS-treated animals and the number of ED2-positive Kupffer cells in tissue sections. The results suggest that ivIG limits the hepatic microvascular inflammatory response during the late phase of sepsis and endotoxemia by affecting Kupffer cell function. PMID- 10847638 TI - Increased extra domain-A containing fibronectin and hepatic dysfunction during septic response: an in vivo and in vitro study. AB - A massive inflammatory reaction resulting from systemic cytokine release is the common pathway underlying sepsis or multiple organ dysfunction. The role of extra domain sequence A-containing fibronectin (EDA+FN) formation during the septic response is not known. The present study investigates the role of EDA+FN during the septic response under in vitro and in vivo conditions. The direct effects of interleukin-1, interleukin-6, and tumor necrosis factor-alpha on EDA+FN production were evaluated in primary cultured human hepatocytes and fibroblasts. Serial plasma EDA+FN levels were measured using an enzyme-linked immunosorbent assay in 24 patients who developed postoperative sepsis following general abdominal surgery of which there were 17 survivors and 7 non-survivors. EDA+FN secretion was significantly increased in cultured hepatocytes but not fibroblasts at 24 and 48 h following exposure to IL-1 compared to controls. In the clinical setting plasma EDA+FN levels in non-survivors were significantly higher than in survivors. Moreover, the EDA+FN levels were correlated closely with liver function tests. EDA+FN levels may represent a specific marker of vascular injury or systemic inflammatory response syndrome that is associated with an adverse clinical outcome. PMID- 10847640 TI - Neurology: reflections on the past millennium as we enter the next. PMID- 10847639 TI - Shock at the millennium. I. Walter B. Cannon and Alfred Blalock. AB - Present management of shock derives, in part, from the classic investigations of Walter B. Cannon and Alfred Blalock. The intersections of their professional lives as recorded in the professional literature and in personal correspondence suggest that Blalock's pivotal studies of experimental shock were fueled, at least in part, by Cannon's inability to resolve the inconsistencies of the then popular toxic theory of shock. Cannon appears to have substantially shaped Blalock's thought and work, initially as authority and competitor and later as colleague and friend. Blalock's experimental proof that injury precipitated obligatory locoregional fluid losses, the effects of which could be ameliorated by vigorous restoration of plasma volume, became a cornerstone of shock theory and therapy. PMID- 10847641 TI - Echoplanar magnetic resonance imaging in acute stroke. AB - Echoplanar magnetic resonance imaging (EPI) enables rapid, non-invasive imaging and analysis of cerebral pathophysiology in acute stroke. It represents an important clinical advance over computed tomography (CT) and conventional magnetic resonance (MR) scanning. It can rapidly delineate infarcted cerebral tissue and distinguish acute from chronic stroke. In addition, EPI has the potential to quickly determine the presence and degree of potentially viable brain tissue in the ischaemic penumbra. Thrombolysis is thought to reperfuse the penumbra and hence reduce infarct size. The thrombolytic agent tissue plasminogen activator (t-PA) improves outcome in ischaemic stroke when administered within the first 3 hours of onset. However, there is a significant risk of haemorrhage, and the time window for benefit may well exceed 3 hours in some patients. Hence, by facilitating diagnosis of 'at-risk' tissue in the ischaemic penumbra, a major clinical role of EPI may well become the rational selection of patients for acute interventional stroke therapy. PMID- 10847642 TI - Aetiological considerations on some conditions in the borderlands of neurology: chronic fatigue syndrome, pan allergy syndrome and repetitive strain injury--a personal view. PMID- 10847643 TI - Reorganisation of the visual cortex in callosal agenesis and colpocephaly. AB - Structural defects involving eloquent regions of the cerebral cortex may be accompanied by abnormal localisation of function. Using functional magnetic resonance imaging (fMRI), we studied the organisation of the visual cortex in a patient with callosal agenesis and colpocephaly, whose visual acuity and binocular visual fields were normal. The stimulus used was a moving grating confined to one hemifield, on a background of moving dots. In addition to activation patterns elicited by stimulation of each hemifield in the patient, the activation pattern was compared to that seen in six normal volunteers. fMRI demonstrated large scale reorganisation of visual cortical areas in the left hemisphere, and fewer activation foci were observed in both occipital lobes when compared with normal subjects. PMID- 10847644 TI - Multiple sclerosis in Australia: prognostic factors. AB - In order to determine the influence of age of onset, sex, onset symptoms, clinical course and interval from onset to first relapse on the subsequent outcome of multiple sclerosis (MS), data from 2934 cases of MS documented in a large population based study undertaken in Australia have been analysed. Disability on prevalence day (30 June 1981) was defined on the Kurtzke disability scale as mild (DSS 0-3), moderate (DSS 4-6) and severe (DSS 7-9). Prognostic factors associated with mild vs moderate/severe, and moderate vs severe disability on prevalence day were identified by logistic regression analysis. A worse prognosis was significantly associated with older age of onset, progressive disease course, onset symptoms that were multiple, pyramidal or cerebellar, and a short interval between onset and first relapse. PMID- 10847645 TI - Spinal cord gliomas: management and outcome with reference to adjuvant therapy. AB - The authors review their experience with 19 consecutive cases with either astrocytic tumour (glioblastoma multiforme one, anaplastic astrocytoma one, astrocytoma 4, pilocytic astrocytoma 4) or ependymoma (10 tumours in 9 patients) of the spinal cord who were treated during the period from 1982 to 1996. The patients included 10 male and 9 female patients with a median age of 38 years. The main tumour locations included the cervicomedullary region 5 the cervical cord (8), the thoracic cord (5) and one each in the thoracolumbar region and conus medullaris. While a total removal of the tumour was achieved in 8 out of 10 ependymomas, the initial treatment for astrocytic tumours was a partial resection in 5, and biopsy in the remaining 5. As adjuvant treatment, 8 patients received radiation therapy and 2 received chemotherapy. Two patients with an astrocytic tumour received chemotherapy only, while the remaining 9 received neither radiation therapy nor chemotherapy initially. After these treatments, 6 out of the 8 patients with low grade astrocytoma have remained alive for 1.3-12.6 years, while 2 patients with high grade astrocytic tumours died within 15 months following surgery. Eight out of 9 patients with an ependymoma have remained alive for 3.0-12.3 years, while one committed suicide 2 years after surgery. As a result, 14 patients are still alive; half of them are accompanied by a mild neurological dysfunction, while the remaining one has a moderate deficit. The postoperative results and the rationale for surgery is discussed, and an approach for utilising adjuvant therapy for high grade tumours is also suggested. PMID- 10847646 TI - Vascular structure of arteriovenous malformations. AB - Cerebral arteriovenous malformations (AVMs) are classified angiographically into two types: the arteriovenous fistula (AVF) type and the plexiform type. However, the differences in vascular structure of these two types have not been clarified. The purpose of the present study is to elucidate the vascular structure of plexiform AVMs and to discuss the clinical significance of this classification of AVMs. Specimens of AVMs resected in 8 cases and identified by cerebral angiography as plexiform AVMs were examined. Immediately after their removal, microdissection of the terminal arterial feeder, the nidus, and the venous drainer was performed under a microscope. A histological examination of each element was then conducted. Microdissection of a portion of the vascular mass that formed the nidus made it possible to separate individual vessels of the mass from each other. Many of these individual vessels connected with the feeder on one side, while the other side anastomosed with the drainer, thus exhibiting the morphology of an AVF. From our examination of the AVMs in the present study, we inferred that the plexiform type is fundamentally a conglomeration of AVFs. It is therefore suggested that the vascular structure of this type of AVM is not fundamentally different from that of the AVF type. PMID- 10847647 TI - Physiological localisation in functional neurosurgery for movement disorders: a simple approach. AB - Controversy exists between anatomical methods and single cell recording as the preferred approach in target localisation in functional neurosurgery for movement disorders. The controversy centres on accuracy as compared to practicality. We describe a mapping technique of semi-microstimulation utilising threshold measurements which has been used in 66 procedures in 50 subjects. We compared the accuracy of anatomical localisation with the final chosen target using the above technique. We also compared the benefit, the side effects and the surgical complication rate with published data on single cell recording and anatomical localisation. The mean difference in 3-dimensional space between the anatomical target and the physiological target was 6.85 mm (P < 0.0001). A good response was obtained in 80% of procedures. Mortality was 1.5%. The surgical complication rate was 1.5%. Mild side effects, serious side effects, transient side effects and permanent side effects were evident in 4.5%, 10.6%, 6.1% and 9.1% of procedures. These figures compared better than anatomical studies and similar to single cell recording studies. It is concluded that this approach provides both accuracy and simplicity and is recommended as a compromise to the currently available methods. PMID- 10847648 TI - Sudden unexpected death in epilepsy in Victoria. AB - Sudden unexpected death in epilepsy (SUDEP) refers to sudden unexpected death in patients with epilepsy in whom autopsy fails to reveal an anatomic or toxicological cause of death. The purpose of this study was to examine associated factors and mechanisms relating to SUDEP in Victoria. The study was a retrospective study based on data from questionnaires completed by treating doctors and coronial files including police reports of death, autopsy and toxicology reports. The deaths were of people with epilepsy in Victoria that were referred to the coroner between 1991 and 1997. There were 15,751 coronial autopsies of which 357 had epilepsy and 50 (14%) were SUDEPs. The SUDEP rate was approximately 1 per 3000 epileptics per year. This study suggested the following associations: young age, tonic-clonic seizures, seizure frequency greater than 10/year, duration of epilepsy greater than 10 years, mental retardation, psychiatric disease and alcohol abuse. Antiepileptic drug (AED) compliance was rated by treating doctors as good in 24 cases. One or more postmortem AED drug levels was subtherapeutic in 30 of 50 cases. Only 5 were receiving psychotropic drugs; only 1 of these was receiving more than one of these drugs. A history of recent unusually stressful life event was present in only 4 cases. At least 11 showed evidence of terminal seizure, and the majority of events occurred in sleep. These observations support the hypothesis that seizures are the mechanism of many cases of SUDEP. The associations observed were largely in agreement with previous studies. However, seizure frequency was greater and duration of epilepsy greater than most previous studies. The role of factors such as AED compliance, psychotropic drug prescription and recent unusually stressful life event is less clear. This highlights the need for case-control studies of risk factors for SUDEP. PMID- 10847649 TI - Huntington's disease: neurological assessment of potential gene carriers presenting for predictive DNA testing. AB - One hundred and fifty-six potential gene carriers who were 50% 'at risk' of inheriting the Huntington's disease (HD) mutation, and who presented for predictive testing, underwent neurological assessment before their gene status had been determined. The association between pre-gene result symptoms and minimal neurological signs (insufficient for diagnosis in their own right) and subsequent gene status was determined. Of these, 38% tested positive for the HD mutation. Fifty-one individuals had minor neurological signs. After exclusions, 61% of gene positive patients had minor neurological signs, whereas only 8% testing gene negative had signs. Minimal chorea observed in the toes and feet with the subject supine, and the patient being stressed by a mental task carried 96% specificity and 86% positive predictive value for gene-positive status. Neurological symptoms did not distinguish gene status, but behavioural and cognitive symptoms were more often reported by the gene-positive group. Although an 'at-risk' individual may receive a gene-positive result, neurological examination remains the most accessible, reliable and cost effective means of determining clinical disease onset. PMID- 10847650 TI - An investigation of the cytotoxicity of the morpholino anthracycline MX2 against glioma cells in vitro. AB - MX2 is a novel morpholino anthracycline reported to have lower systemic toxicity than other anthracyclines. It has similar antitumour activity to adriamycin and is cytotoxic towards multi-drug resistant cells and anthracycline sensitive sublines of human and murine tumour cells. In this study MX2 showed a marked cytocidal effect compared to M2, the most cytotoxically active metabolite, and the nitrosourea, BCNU, when 30 ng/ml of each drug was added to separate flasks of C6 glioma cells grown in monolayer. The colony formation of C6 glioma cells was markedly inhibited by MX2 in a dose dependent manner. The LD50 values for MX2, M2 and BCNU were 10.5 ng/ml, 15.8 ng/ml and 465 ng/ml respectively. MX2 is likely to be bound to the main plasma protein, albumin, and can also interact with the plasma lipoproteins, particularly high density lipoprotein. The results in this study strongly support the further investigation of MX2 as a potential chemotherapeutic agent against brain tumours. PMID- 10847652 TI - Computer generated microsurgical anatomy of the supraclinoid portion of the internal carotid artery. AB - In this article we demonstrate a computer graphics model to represent the microsurgical anatomy of the supraclinoid portion of the internal carotid artery (ICA). The source of the input data is a variety of publications showing the detailed anatomy of the area. A computer graphics model of the supraclinoid portion of the ICA and relevant structures including perforators and cranial nerves was made under full colour shading using a Z-buffer algorithm. Complicated microstructures including perforators or cranial nerves are depicted in detail, and can be rotated and viewed from any direction, and thus allows understanding of the three-dimensional relations of the area. Using our method, it may be possible to represent more complicated models of various anatomies, and this methodological tool would be useful for teaching surgical microanatomy and pathology and in pre-surgical operative planning and preparation of the surgeon, introducing a new style navigational system for open microneurosurgery. PMID- 10847651 TI - Chronic effects of an aminosteroid on microdialytically measured parameters after experimental middle cerebral artery occlusion in the rat. AB - The effects of the neuroprotective aminosteroid U74006F (tirilazad mesylate, Freedox) were monitored microdialytically in rat cortex during three 4h periods beginning immediately, 25h and 49h after permanent middle cerebral artery occlusion. Either U74006F or vehicle only was administered 15 min, 2h, 6h, 12h and 24h after operation. The dialysate was analysed for on-line pH, ascorbic acid, uric acid, glucose and lactate. The efficacy of post-ischaemic treatment was shown by: a) lesion volume 53h after operation was significantly smaller in U74006F-treated animals; b) microdialytic findings were very similar to those found previously with pre-ischaemic drug application (reduction in release of ascorbic acid, uric acid and lactate, increased pH); c) an effect of U74006F on lactate release could still be seen on days 2 and 3; and d) increases in uric acid on days 2 and 3, possibly reflecting delayed cell death, were smaller in aminosteroid treated animals. PMID- 10847653 TI - A SPECT study of the anatomy of transient global amnesia. AB - This report describes single photon emission tomography (SPECT) evidence of transient, relative hypoperfusion of the right basal ganglia and left temporal lobe in a 60-year-old man studied during the acute phase of classical transient global amnesia (TGA). The findings are interpreted in the light of previous reports indicating that involvement of the right hemisphere or both cerebral hemispheres consistently predicts severe retrograde amnesia, in addition to the typical loss of anterograde memory observed during TGA. These deficits involving the right hemisphere and left temporal lobe may reflect disruption of circuits mediating episodic memory retrieval and memory encoding, respectively. PMID- 10847654 TI - HELLP syndrome with haemaglobin vasospasm. AB - The syndrome of haemolysis, elevated liver enzymes and low platelets (HELLP syndrome) is a life threatening, severe complication of pre-eclampsia with typical laboratory findings. An unusual case of a 36-year-old woman with HELLP syndrome and the initial complication of intracerebral haemorrhage is presented. The diagnosis of HELLP syndrome was confirmed by elevated liver enzymes, low platelets, increased total bilirubin and increased lactate dehydrogenase. The intracranial haematoma was removed with good neurological recovery ensuing. However, this case was complicated by cerebral vasospasm on the eleventh day, confirmed by cerebral angiography and computer tomographic findings. The patient died from brain swelling. Possible vasospam should be considered during the treatment of patients with HELLP syndrome. PMID- 10847655 TI - Isolated superior division oculomotor palsy in a child with spontaneous recovery. AB - A 10 year old boy with a superior division palsy of the left oculomotor nerve is reported. He had a flu-like illness 1 week before the onset. The computed tomography and magnetic resonance imaging scans were normal. Laboratory data for evaluation of infection, diabetes mellitus and myasthenia gravis were normal. The symptoms spontaneously disappeared after 2 months. The course of the illness in conjunction with the negative laboratory findings made this case an example of partial paralysis of the third nerve related to viral infection. In the literature, only several cases with isolated divisional palsy of the oculomotor nerve were found after a viral infection. Inferior division palsy has been reported in five children. Superior division palsy has been published in only two cases. Divisional palsy is more common among children and resolves spontaneously. This rare but important clinical entity is one of the differential diagnoses in oculomotor nerve palsies, particularly in children, which are neuroradiologically undiagnosed. It occurs after a viral infection and may affect a superior or inferior division alone. PMID- 10847656 TI - Spinal dural arteriovenous fistula with perimesencephalic subarachnoid haemorrhage. AB - A case is reported of a 66 year old woman presenting with perimesencephalic subarachnoid haemorrhage (SAH) which was caused by a spinal dural arteriovenous fistula at the C1 level. The fistula drained into the venous system of the posterior cranial fossa through a perimedullary vein. The bleeding was thought to result from venous hypertension induced by the fistula. This case may support the hypothesis that perimesencephalic non-aneurysmal SAH can be ascribed to venous bleeding and that venous hypertension is the key to its pathology. PMID- 10847657 TI - Arachnoiditis ossificans and syringomyelia: a unique case report. AB - A 62-year-old male presented with progressive quadriparesis. Magnetic resonance imaging of the spine revealed a spinal cord syrinx but failed to detect extensive arachnoiditis ossificans noted on insertion of a syringopleural shunt. A postoperative computed tomography scan clearly demonstrated the abnormality and its extent. We present a rare case of syringomyelia resulting from spinal arachnoiditis ossificans and review the relevant literature. PMID- 10847658 TI - Metastatic atypical meningioma: case report and review of the literature. AB - Extracranial metastasis of an intracranial meningioma is rare. We discuss the clinical, radiological and histopathological presentation of an elderly man with pulmonary metastases from a recurrent meningioma of atypical histology, and review the literature pertaining to this phenomenon. PMID- 10847659 TI - Nitrous oxide myelopathy in an abuser of whipped cream bulbs. AB - A 23 year old man presented with a severe posterior column myelopathy related to prolonged nitrous oxide abuse obtained from whipped cream bulbs. The site of pathology was identified by magnetic resonance imaging (MRI) and somatosensory evoked potentials. The mechanism of toxicity involves inactivation of vitamin B12 dependent enzymes. Appropriate treatment with methionine and vitamin B12 was instituted quickly with good neurological outcome. There are major concerns regarding the availability of nitrous oxide in supermarkets. PMID- 10847660 TI - In favour of lumbar fusion. PMID- 10847661 TI - Lumbar fusion--the case against. PMID- 10847662 TI - From Scandinavian Journal of Social Medicine to Scandinavian Journal of Public Health: a change of name but not of vision. PMID- 10847663 TI - Need for medical care and social services. AB - Predicting the need for care in the coming decades requires direct measurement of need in birth cohorts, allowing for differentiation with respect to age, generation effects, and the effect of period of time. The method of choice is a simulation of regular contacts with the services, implicit criteria of need for service, and a panel approach with measurement at three points in time. PMID- 10847664 TI - Vocational rehabilitation of the socially disadvantaged long-term sick: inter organizational co-operation between welfare state agencies. AB - Vocational rehabilitation targeted to the socially disadvantaged long-term sick requires that the client keep in touch with a number of welfare state agencies, all of which have different regulations, conflicting goals and various types of benefits. This is an arduous and time-consuming task for clients with medical, social and labour market problems. Many of these clients run the risk of ending up in a no-man's land or being endlessly circulated between agencies because their problems do not correspond to the profile of the typical client. Both government and welfare workers see institutional co-operation between welfare state agencies as the remedy to such problems. This article, which is based on interviews with participants in fourteen cooperating projects, focuses on difficulties and opportunities experienced in such co-operation. It is concluded that such cooperation, when initiated in local settings and supported by local players, is a way of rejuvenating the existing Swedish model. PMID- 10847665 TI - The importance of self-efficacy in cardiovascular risk factor change. AB - SUBJECTS AND DESIGN: The study was based on the total patient sample (n = 110) of a randomized controlled trial comparing two intervention methods advising cardiovascular high-risk men of lifestyle changes in general practice. Behaviour and risk factor changes during the one-year intervention study were analysed using multiple regression and logistic regression analyses with the above mentioned independent variables. SETTING: Twenty-two general practice centres in the county of Hordaland, western Norway. RESULTS: Self-efficacy of increased physical exercise was the only variable significantly related to exercise change. Age and self-efficacy were statistical significant predictors of smoking cessation success. None of the independent variables was statistically significantly related to blood pressure or cholesterol change. Educational level related negatively, although statistically insignificantly, with total risk change. CONCLUSION: The study confirms the importance of self-efficacy in both human behaviour and motivation for behaviour change. OBJECTIVES: The objectives of the study were to explore the impact of possible predictors for cardiovascular risk behaviour change, predictors such as education, age, self-efficacy, doctors' interpersonal skills, and number of appointments. PMID- 10847666 TI - Physicians' legal defensiveness and knowledge of medical law: comparing Denmark and the USA. AB - The impact of legal factors on medical treatment decisions for dying patients has been studied in the USA for years. However, it is unknown how legal factors may affect similar medical decisions in other countries. This exploratory study compared responses between two populations of physicians, from Denmark (n = 62) and the USA (n = 301), who regularly treat terminally ill patients in tertiary care medical centers. We investigated whether Danish and US physicians differed significantly in their attitudes about the influence of law on treatment decisions for terminally ill patients. The Danish physicians demonstrated significantly better knowledge of medical law relevant to end-of-life treatment than did US physicians. The Danish sample also reported significantly lower levels of legal defensiveness than the US sample. These findings are consistent with our previous research showing that, among US physicians, legal defensiveness and knowledge of medical law are inversely related. PMID- 10847667 TI - Swedish physicians' inclination to prescribe benzodiazepines: differences between regions and characteristics of the prescriber. AB - There are substantial differences in the sales of benzodiazepines between Swedish regions. In order to determine whether there are consistent differences in the inclination of Swedish doctors to prescribe benzodiazepines depending on region and characteristics of the prescriber, a questionnaire was sent to a stratified random sample of physicians (116 family practitioners, 78 psychiatrists, and 96 private practitioners). The questionnaire contained six case studies in which benzodiazepine treatment was suggested and two case studies concerning change of benzodiazepine dosage. There were significant differences (p < 0.05) in the inclination to prescribe benzodiazepines depending on both region and the physician's age in two cases and the physician's speciality and gender in one case each. Psychiatrists and female physicians were, in general, less inclined to prescribe benzodiazepines. In spite of higher sales of benzodiazepines in Goteborg, physicians there had a lower inclination to prescribe benzodiazepines compared with physicians working in regions with lower prescription rates. PMID- 10847668 TI - Women's alcohol dependence and abuse: the relation to social network and leisure time. AB - The aim of this study was to analyse the occurrence of alcohol dependence and abuse in relation to variables reflecting social network and leisure time, social class, education, occupation and family conditions. The study was based on the second wave of interviews performed within the longitudinal study of Women and Alcohol in Goteborg. In this present study we analysed 416 face-to-face interviews performed in 1995 96. Women with only one or no friends for support had higher rates of alcohol dependence and abuse than did women with more friends. Women with high rates of alcohol dependence or abuse did not take part in cultural events as much as women with low rates of alcohol dependence and abuse. Women with homemaking skills and gardening as leisure time interests had lower prevalences of alcohol dependence and abuse. In general, women with diagnoses of alcohol dependence and abuse in this population-based sample were not alone and without a social network. However, their pattern of activity differed slightly from those without such diagnoses. The association between women with alcohol dependence and abuse and leisure time activities is probably circular: a poor social network and low participation in social activities increase the risk of alcohol dependence and abuse, and alcohol dependence and abuse lead to low participation in social activities. PMID- 10847669 TI - Health-related quality of life in patients with diastrophic dysplasia. AB - Diastrophic dysplasia (DD), a congenital skeletal dysplasia, is characterized by short, disproportionate stature, multiple severe spinal and joint deformities, and normal mental status. The health-related quality of life (HRQOL) of patients with DD was measured by a standardized fifteen-dimensional (15D) method, that includes a questionnaire and a valuation task. Eighteen patients (5 males and 13 females) with a mean age of 23 years (range 17-31 years), representing half of the Finnish patients with DD in this age group, completed the questionnaire and the valuation task. The data obtained were compared with those of 273 age- and sex-matched controls. A 15D profile was drawn, and the average importance weight of each dimension and the average within-dimension level values were calculated for both groups. The average 15D score, describing the overall HRQOL, was derived for both groups using the evaluations of each group. The health profiles of the groups differed significantly in several dimensions. When the health levels within the dimensions were concerned, the controls rated all values deviating from "normal" lower than did the patients. The average 15D scores of the patients with DD were significantly lower than those of the controls. The 15D scores were higher in both groups when the evaluations of the patients with DD were used. The study showed that, although the overall HRQOL of the patients with DD was inferior to that of the controls, the patients showed greater adaptability to deviations in health status than did the controls. PMID- 10847670 TI - Socioeconomic differences in health and well-being of children and adolescents in Iceland. AB - STUDY OBJECTIVE: To assess differences in health, healthcare use and well-being of children according to their socioeconomic situation. DESIGN: Part of a larger cross-sectional survey on health and well-being of children and their parents in the Nordic countries. SETTING: Iceland. PARTICIPANTS: A questionnaire was sent to the parents of a nationally representative sample of 3,007 school children aged two to seventeen years. MAIN RESULTS: The SES indicators used were education and occupation of both parents and the disposable income of the family. Logistic regression models were used for the analysis. Children of lower SES were found to have worse health and well-being than those of higher SES. Children of lower SES appeared to use doctor's services to the same degree as children of higher SES, especially after differences in ill health were taken into account. CONCLUSION: The association between SES and health status and well-being in adulthood can already be detected in childhood, even in an egalitarian country with a homogeneous population. PMID- 10847671 TI - Drug-related mortality in Denmark 1970-93. AB - We analysed drug-related mortality in Denmark with respect to secular trends, gender, and regional variations, for the period 1970-93, for all deaths from poisoning and among drug addicts. The study was based on the Register of Causes of Death in Denmark and included 6,229 drug-related deaths, defined by specific combinations of manner of death, underlying cause of death, and contributory cause of death. The main outcome measure is age-specific mortality rate. A total of 63% of the drug-related deaths were registered as unnatural deaths. During the period studied, mortality increased for men in the 25 49 year age group and for women in all age groups over 25 years of age. For both men and women, the youngest birth cohorts from the mid-1950s and 1960s suffered much higher mortality than those born before 1950; however, the three youngest birth cohorts had almost the same mortality. During the entire period, mortality in the capital, Copenhagen, was much higher than in the provinces, but in the last years, a more favourable trend has been seen in Copenhagen. PMID- 10847672 TI - Pregnancy-related violent deaths. PMID- 10847673 TI - Are male and female smokers at equal risk of smoking-related cancer: evidence from a Swedish prospective study. AB - This study examines sex differences in the relative risks of lung cancer and other smoking-related cancers (i.e. cancers of the upper respiratory tract, oesophagus, pancreas, bladder, and renal pelvis). Data on smoking habits in 1963 from a random sample of 56,000 men and women were linked with information on new cases of cancer for 1964-89. Compared with people who have never smoked, the relative risks of lung cancer at different levels of pack-years completed in 1963 (>5, 6-15, 16-25 and 25 + pack-years) were 1.6, 4.4, 14.2, and 17.9 for men, and 2.1, 6.3, 10.3, and 16.5 for women. The corresponding relative risks of other smoking-related cancers were 1.8, 3.0 5.4, and 6.4 for men, and 2.0, 3.1, 5.0, and 6.5 for women. These results suggest that men and women have similar relative risks of smoking-related cancers at different levels of smoking. PMID- 10847674 TI - A scoring system for subjective health complaints (SHC). AB - The aim of this study is to present a complete scoring system for subjective health complaints (SHC) as they are experienced by the lay population. The scoring system records the complaints, and does not map attributions or medical diagnoses. In all, 1,219 subjects (323 men, 896 women) from various occupations were tested with a scoring system, the SHC inventory, previously referred to as the Ursin Health Inventory (UHI). The SHC consists of 29 questions concerning severity and duration of subjective somatic and psychological complaints. The SHC inventory yields scores on single items and a total number of health complaints categorized into five factors: musculoskeletal pain (alpha=0.74), pseudoneurology (alpha=0.73), gastrointestinal problems (alpha=0.62), allergy (alpha=0.58) and flu (alpha=0.67). The SHC inventory is a systematic, easy, and reliable way to score subjective health complaints. The prevalence of these complaints is high, and should be considered before conclusions are reached about new diseases and new attributions of environmental hazards. PMID- 10847675 TI - Validity of two questions on alcohol use in a health survey questionnaire. AB - The aim of this study was to investigate whether consumers of high and low levels of alcohol could be identified by two questions about alcohol use in a postal questionnaire survey. A sample of 2,300 persons aged 18 64 years from Stockholm county were sent a masked postal questionnaire comprising 30 questions about their health and functioning. Two questions concerned their alcohol consumption. One year later the subjects underwent a psychiatric health examination, which included an assessment of their alcohol use. The two questions about alcohol consumption identified high alcohol consumers with a relatively high sensitivity and specificity, of 64% and 87%, respectively, and thus are useful for identifying high alcohol consumers in health surveys using questionnaires. PMID- 10847676 TI - What is a population-based registry? PMID- 10847677 TI - Double-stranded RNA as a template for gene silencing. PMID- 10847678 TI - Orchestral maneuvers in the axon: trio and the control of axon guidance. PMID- 10847679 TI - Cell cycle-dependent translation initiation: IRES elements prevail. PMID- 10847680 TI - Functional and genomic analyses reveal an essential coordination between the unfolded protein response and ER-associated degradation. AB - The unfolded protein response (UPR) regulates gene expression in response to stress in the endoplasmic reticulum (ER). We determined the transcriptional scope of the UPR using DNA microarrays. Rather than regulating only ER-resident chaperones and phospholipid biosynthesis, as anticipated from earlier work, the UPR affects multiple ER and secretory pathway functions. Studies of UPR targets engaged in ER-associated protein degradation (ERAD) reveal an intimate coordination between these responses: efficient ERAD requires an intact UPR, and UPR induction increases ERAD capacity. Conversely, loss of ERAD leads to constitutive UPR induction. Finally, simultaneous loss of ERAD and the UPR greatly decreases cell viability. Thus, the UPR and ERAD are dynamic responses required for the coordinated disposal of misfolded proteins even in the absence of acute stress. PMID- 10847681 TI - Structure of the ERM protein moesin reveals the FERM domain fold masked by an extended actin binding tail domain. AB - The ezrin-radixin-moesin (ERM) protein family link actin filaments of cell surface structures to the plasma membrane, using a C-terminal F-actin binding segment and an N-terminal FERM domain, a common membrane binding module. ERM proteins are regulated by an intramolecular association of the FERM and C terminal tail domains that masks their binding sites. The crystal structure of a dormant moesin FERM/tail complex reveals that the FERM domain has three compact lobes including an integrated PTB/PH/ EVH1 fold, with the C-terminal segment bound as an extended peptide masking a large surface of the FERM domain. This extended binding mode suggests a novel mechanism for how different signals could produce varying levels of activation. Sequence conservation suggests a similar regulation of the tumor suppressor merlin. PMID- 10847683 TI - Trio combines with dock to regulate Pak activity during photoreceptor axon pathfinding in Drosophila. AB - Correct pathfinding by Drosophila photoreceptor axons requires recruitment of p21 activated kinase (Pak) to the membrane by the SH2-SH3 adaptor Dock. Here, we identify the guanine nucleotide exchange factor (GEF) Trio as another essential component in photoreceptor axon guidance. Regulated exchange activity of one of the two Trio GEF domains is critical for accurate pathfinding. This GEF domain activates Rac, which in turn activates Pak. Mutations in trio result in projection defects similar to those observed in both Pak and dock mutants, and trio interacts genetically with Rac, Pak, and dock. These data define a signaling pathway from Trio to Rac to Pak that links guidance receptors to the growth cone cytoskeleton. We propose that distinct signals transduced via Trio and Dock act combinatorially to activate Pak in spatially restricted domains within the growth cone, thereby controlling the direction of axon extension. PMID- 10847682 TI - act up controls actin polymerization to alter cell shape and restrict Hedgehog signaling in the Drosophila eye disc. AB - Cells in the morphogenetic furrow of the Drosophila eye disc undergo a striking shape change immediately prior to their neuronal differentiation. We have isolated mutations in a novel gene, act up (acu), that is required for this shape change. acu encodes a homolog of yeast cyclase-associated protein, which sequesters monomeric actin; we show that acu is required to prevent actin filament polymerization in the eye disc. In contrast, profilin promotes actin filament polymerization, acting epistatically to acu. However, both acu and profilin are required to prevent premature Hedgehog-induced photoreceptor differentiation ahead of the morphogenetic furrow. These findings suggest that dynamic changes in actin filaments alter cell shape to control the movement of signals that coordinate a wave of differentiation. PMID- 10847684 TI - Single active site catalysis of the successive phosphoryl transfer steps by DNA transposases: insights from phosphorothioate stereoselectivity. AB - The transposase family of proteins mediate DNA transposition or retroviral DNA integration via multistep phosphoryl transfer reactions. For Tn10 and phage Mu, a single active site of one transposase protomer catalyzes the successive transposition reaction steps. We examined phosphorothioate stereoselectivity at the scissile position for all four reaction steps catalyzed by the Tn10 transposase. The results suggest that the first three steps required for double strand cutting at the transposon end proceed as a succession of pseudo-reverse reaction steps while the 3' end of the transposon remains bound to the same side of the active site. However, the mode of substrate binding to the active site changes for the cut transposon 3' end to target DNA strand joining. The phosphorothioate stereoselectivity of the corresponding steps of phage Mu transposition and HIV DNA integration matches that of Tn10 reaction, indicating a common mode of substrate-active site interactions for this class of DNA transposition reactions. PMID- 10847685 TI - A chromodomain protein, Swi6, performs imprinting functions in fission yeast during mitosis and meiosis. AB - Inheritance of stable states of gene expression is essential for cellular differentiation. In fission yeast, an epigenetic imprint marking the mating-type (mat2/3) region contributes to inheritance of the silenced state, but the nature of the imprint is not known. We show that a chromodomain-containing Swi6 protein is a dosage-critical component involved in imprinting the mat locus. Transient overexpression of Swi6 alters the epigenetic imprint at the mat2/3 region and heritably converts the expressed state to the silenced state. The establishment and maintenance of the imprint are tightly coupled to the recruitment and the persistence of Swi6 at the mat2/3 region during mitosis as well as meiosis. Remarkably, Swi6 remains bound to the mat2/3 interval throughout the cell cycle and itself seems to be a component of the imprint. Our analyses suggest that the unit of inheritance at the mat2/3 locus comprises the DNA plus the associated Swi6 protein complex. PMID- 10847686 TI - ZEITLUPE encodes a novel clock-associated PAS protein from Arabidopsis. AB - We have conducted genetic screens for period length mutants in Arabidopsis using a transgenic bioluminescence phenotype. This screen identified mutations at a locus, ZEITLUPE (ZTL), that lengthen the free-running period of clock-controlled gene transcription and cell expansion, and alter the timing of the daylength dependent transition from vegetative to floral development. Map-based cloning of ZTL identified a novel 609 amino acid polypeptide consisting of an amino-terminal PAS domain, an F box and six carboxy-terminal kelch repeats. The PAS region is highly similar to the PAS domain of the Arabidopsis blue-light receptor NPH1, and the Neurospora circadian-associated protein WHITE COLLAR-1 (WC-1). The striking fluence rate-dependent effect of the ztl mutations suggests that ZTL plays a primary role in the photocontrol of circadian period in higher plants. PMID- 10847688 TI - The potassium salt of p-nitrocatechol sulfate. PMID- 10847687 TI - FKF1, a clock-controlled gene that regulates the transition to flowering in Arabidopsis. AB - Plant reproduction requires precise control of flowering in response to environmental cues. We isolated a late-flowering Arabidopsis mutant, fkf1, that is rescued by vemalization or gibberellin treatment. We positionally cloned FKF1, which encodes a novel protein with a PAS domain similar to the flavin-binding region of certain photoreceptors, an F box characteristic of proteins that direct ubiquitin-mediated degradation, and six kelch repeats predicted to fold into a beta propeller. FKF1 mRNA levels oscillate with a circadian rhythm, and deletion of FKF1 alters the waveform of rhythmic expression of two clock-controlled genes, implicating FKF1 in modulating the Arabidopsis circadian clock. PMID- 10847689 TI - Myocardial acute and chronic histological modifications induced by cryoablation. PMID- 10847690 TI - Alleged misconduct in breast cancer trial. PMID- 10847691 TI - Testicular cancer gene located. PMID- 10847692 TI - HRT 'reduces mammogram sensitivity'. PMID- 10847693 TI - Radiotherapy in non-invasive breast cancer. PMID- 10847694 TI - Mathematicians model cancer growth. PMID- 10847695 TI - Managing psychological morbidity in cancer patients. PMID- 10847696 TI - New guidelines to evaluate tumour response. PMID- 10847697 TI - Cancer charter signed in Paris. PMID- 10847698 TI - Exercise 'decreases breast cancer risk'. PMID- 10847700 TI - Cancer genome project to start. PMID- 10847699 TI - Young breast cancer patients 'should be offered chemotherapy'. PMID- 10847701 TI - Ovarian cancer treatment 'still ignored'. PMID- 10847702 TI - Low-grade gastric MALT lymphoma as a second malignancy in chronic lymphocytic leukaemia. PMID- 10847703 TI - Circadian variation of granulocyte colony-stimulating factor levels in man. PMID- 10847704 TI - Angiogenic growth factors and endostatin in non-Hodgkin's lymphoma. PMID- 10847705 TI - Ala-10 mutations in the factor IX propeptide and haemorrhage in a patient treated with warfarin. PMID- 10847706 TI - Conjunctival tumour as the primary manifestation of infectious mononucleosis in a 12 year old girl. PMID- 10847707 TI - Ultrasound biomicroscopy in the diagnosis of a foreign body simulating iris melanoma. PMID- 10847708 TI - Optical coherence tomography findings in early solar retinopathy. PMID- 10847709 TI - Granulomatous uveitis, CREST syndrome, and primary biliary cirrhosis. PMID- 10847710 TI - Vitreous basket sign in dislocation of the lens. PMID- 10847711 TI - Combined central retinal vein occlusion and cilioretinal artery occlusion in a patient on hormone replacement therapy. PMID- 10847712 TI - Histological examination of the pigment epithelium-Bruch membrane-chorio capillaris complex after macular translocation. PMID- 10847713 TI - Optic neuropathy as the presenting feature of HIV infection: recovery of vision with highly active antiretroviral therapy. PMID- 10847714 TI - With regard to the relative merits of contemporary measurements and historical calculated fields in the Swedish Childhood Cancer Study. PMID- 10847715 TI - The 4th Annual Symposium on Radiotherapy to Reduce Restenosis. La Jolla, California, USA. January 13-15, 2000. Abstracts. PMID- 10847716 TI - Radiation therapy in exudative age-related macular degeneration. AB - It has been suggested that ionizing radiation at doses relatively safe to the optic nerve and retina exert an inhibitory and occlusive effect on the endothelial proliferation of choroidal neovascularization (CNV) associated with age-related macular degeneration (AMD). The encouraging results of early studies in preservation or improvement of visual acuity and regression of the CNV gave rise to many clinical trials in different centers. Disparate radiation doses, dose fractions, type and rate of radiation administration have been used to determine the efficacy of radiotherapy in AMD. Conflicting treatment responses have been reported by different centers. Some studies provided evidence of beneficial treatment outcome in AMD, and others could not show any efficacy of ionizing radiation in the visual and morphological evolution of the disease. Data from the literature and our experience indicate that radiotherapy can be effective in regressing the leakage of the CNV in AMD. However, despite treatment visual deterioration continues and new CNV lesions develop. The observation of morphological progression in the disease process might be related to an unfavorable effect of radiation on the pathogenesis of AMD. PMID- 10847717 TI - [Record no. 22: Beta-catenin]. PMID- 10847718 TI - [Record no. 23: hMLH1]. PMID- 10847719 TI - [Cancer of the uterine cervix: combination of radiotherapy and cisplatin chemotherapy (chemoradiotherapy) is superior to radiotherapy alone in local advanced stages]. PMID- 10847720 TI - [The Blm(-/-) mice: an stage towards the understanding the molecular mechanisms at play in Bloom syndrome]. PMID- 10847721 TI - [c-Myc: an iron oncogene]. PMID- 10847722 TI - [Could a deficit in the expression of the ATM gene explain its role in the incidence of cancer?]. PMID- 10847723 TI - [The Wnt1 oncogene: from mice mammary tumors to human tumors (continuation)]. PMID- 10847724 TI - [When hMLH1 is implicated in cancers other than HNPCC]. PMID- 10847725 TI - [MEN-1-jun D: an association of benefactors?]. PMID- 10847727 TI - International Conference on Periodontal Research. Rochester, New York, August 1 3, 1969. Research Reports. PMID- 10847726 TI - Second International Conference on Periodontal Research. Scahticon, Aarhus, Denmark, August 2-4, 1972. Research Reports. PMID- 10847728 TI - Electron microscopic study of chronic desquamative gingivitis. PMID- 10847729 TI - Commentary: delay in stopping treatment can become unreasonable and unfair. PMID- 10847730 TI - Fallout from the Shipman case. Existing safeguards against secret homicide are defective and have been weakened. PMID- 10847731 TI - Fallout from the Shipman case. A system is already in place, but the powers of medical referees in cremation must be made truly effective. PMID- 10847732 TI - Fallout from the Shipman case. Death registers in general practice would be a means of preventing malpractice and murder. PMID- 10847733 TI - Fallout from the Shipman case. Publication of mortality data for individual GPs will keep focus on potential to do harm. PMID- 10847734 TI - Fallout from the Shipman case. Group practice safeguards patients. PMID- 10847735 TI - Fallout from the Shipman case. Having any social elite has dangers, but beware the pitfalls in regulating professionals. PMID- 10847736 TI - Fallout from the Shipman case. Requirement that general practice records be on government forms for legal reasons needs to be updated. PMID- 10847737 TI - Fallout from the Shipman case. Hasty political decision on serious professional misconduct may restrict GMC's regulatory abilities. PMID- 10847738 TI - Fallout from the Shipman case. Shipman's family should not have to face penury as well as everything else. PMID- 10847739 TI - Has the sun protection factor had its day? Sunscreen users need education in application technique. PMID- 10847740 TI - The ethics of unlinked anonymous testing. Testing need not stop. PMID- 10847741 TI - Language, fairness, and the MRCGP examination. What is important is who will make a good doctor. PMID- 10847742 TI - More on the facelift flap. PMID- 10847743 TI - Regarding current issues in dermatologic office based surgery. PMID- 10847744 TI - A simple technique for the correction of excessively projectile nipples. PMID- 10847745 TI - Special issue: Electron microscopy of aperiodic materials. PMID- 10847746 TI - [Slovak Surgical Society and the 1st Surgical Clinic of FN and LFUK in Bratislava. The 52nd Kostlivy Surgery Seminar. Collection of work presented on modern surgery and infection. Bratislava 1998]. PMID- 10847747 TI - [Bacterial resistance to antibiotics in surgery: summary of the antibiotic era]. PMID- 10847748 TI - [Antibiotic prophylaxis in surgery]. AB - Infection remains a serious complication after surgical produces. The main risk factors for developing infection are: 1. endogenous-host related, 2. exogenous produce related, 4. environmental-related ones. Systemic antibiotic prophylaxis significantly reduces the incidence of potentially serious infective complications. It is indicated in procedures with incidence of infective complications more as 5%, in clean contaminated wounds and also in produces, in which infection has fatal consequences (vascular surgery, heart surgery, traumatology). The decision to use antibiotic prophylaxis depends upon the operation to be performed, the findings at operations, the general health of the patient and pharmacological and antibacterial properties of the agent. Timing of the first dose (administration not more as 1 hour preoperatively) and duration not more as 24 hours are very important. We use the second generations of cephalosporins (cefuroxim and after antibiotic rotation cefamandol) in antibiotic prophylaxis obligatory in vascular surgery, pacemaker implantation, traumatology and in colorectal surgery (there in combination with metronidasol) with mean infection rate in clean surgical procedures from 0.5 to 1.5%. Complications after antibiotic prophylaxis are very rare. However antibiotic prophylaxis can not compensate the correction of medical problems preoperatively and the meticulous surgical technique. PMID- 10847749 TI - [Infectious complications after laparoscopic procedures on the biliary tract]. PMID- 10847750 TI - [Early infectious complications in gallbladder surgery then and now]. PMID- 10847751 TI - ["One shot" antibiotic prophylaxis in abdominal surgery]. PMID- 10847752 TI - [Inflammatory complications in reconstructive surgery after esophagectomy]. AB - The authors evaluated in their retrospective study the portion of inflammatory complications on morbidity and mortality at restorative operations after oesophagectomies, performed at Ist Department of Surgery of the Teaching Hospital of Commenius University in Bratislava in the year of 1975-1998. The inflammatory complications still remain a serious problem after oesophagectomy with a great portion on postoperative mortality. PMID- 10847753 TI - [Current problems with infections in thoracic surgery]. PMID- 10847754 TI - [Prevention and treatment of prosthesis-related sepsis in vascular surgery]. AB - BACKGROUND: The first synthetic vascular prosthesis have been implanted 47 years ago. 5 years after have been referred the first aortoenteric fisula in connection with the synthetic vascular prosthesis implantation. MAIN PURPOSE AND STARTING POINTS (OBJECTIVES): The incidence of the infection and septic postoperative complications after vascular surgery was relatively high in the past. Several authors note a reduction in the incidence with experience. Another change has been widely noted recently concerning the incidence of various strains of bacteria isolated from the infected tissue and infected prostheses. The most common strain at present is St. epidermidis, known in the past as a saprophyte. There have been also changed the methods of diagnosis and the management policy. The authors of the present paper has been tried to summarize the knowledge in this field on the background of their own experience. They have summarized the clinical classification, bacteriology and the pathophysiologic mechanism of the infection, the most common clinical features, basic diagnostic possibilities, prophylactic and therapeutic policy in the postoperative septic complications, as well. PMID- 10847755 TI - [Infected arteriovenous fistulae for hemodialysis]. AB - Angioaccess procedures for hemodialysis over a 12-year period were retrospectively reviewed to ascertain the frequency of infectious complications. A total of 571 angioaccess procedures were performed including 88 e polytetrafluoroethylene (GORE) grafts. Early infectious complications occurred only in 5 autogenous fistulae. Late infectious complications occurred in 2.3% autogenous fistulae and in 7.95% ePTFE grafts. Infected arteriovenous fistula can be the cause of further complications (thrombosis, bleeding, aneurysm, sepsis) and demands surgical repair. PMID- 10847756 TI - [Use of sulperazone in acute surgical conditions]. AB - In the period between February 2, 1988 and October 30, 1998, the investigators administered Sulperazon to 20 patients at the Surgery Department 1, University Hospital Olomouc, in whom they expected good clinical effect (i.e., high concentration in bill, effect on respiratory tract pathogens). Sulperazon was administered in 12 cases as a drug of first choice, in 8 cases as a drug of second choice. In all cases, relief from clinical symptoms of infection was observed within 5 days of treatment and in 83% of cases eradication of previously cultivated pathogen was achieved. Sulperazon was well tolerated and no side effects or impoivment of laboratory test were observed. In 4 cases, prolonged administration was complicated by candida superinfection. Sulperazon proved to be effective in surgery in treatment of infectious complications of bile duct and pancreas diseases and in thoracic cavity pathology. PMID- 10847757 TI - [[Antibiotic prophylaxis in extensive reconstruction of the esophagus in carcinoma]. PMID- 10847758 TI - [Late infections of aortic prostheses]. AB - The authors present their experience in late infection of aortoilliacofemoral reconstruction by means of the Dacron vascular prosthesis. They found such complication in 8 of 620 (1.5%) reconstructions in that region in period 1988 1998. There is essential position of scintigraphy (Le 99mTc-HMPAO) and CT in diagnostic of location and extent of an affection. Conservative approach and local surgery were not permanently successful. The alternative approach is radical removing of the whole graft. The alternative of aortofemoral reconstruction is axillobifemoral bypass using ePTFE graft in case of an infection of the bifurcation graft. PMID- 10847759 TI - Current views on surgical treatment of pancreatic cancer. PMID- 10847760 TI - Reflux symptoms: can we sort them out? PMID- 10847761 TI - Bioluminescent fusion conjugates and bioluminescent immunoassays: 1988-1998. PMID- 10847763 TI - [Cell adhesion molecules and autoimmune nervous system diseases, with special reference to beta 1 integrin molecule and its signal molecule Cas-L]. PMID- 10847762 TI - Time-resolved fluorescence: 1996-1998. PMID- 10847764 TI - Chemokine receptors and other surface molecules preferentially associated with human Th1 or Th2 cells. PMID- 10847765 TI - Reprogramming transcription during the differentiation of precursor CD4+ T cells into effector Th1 and Th2 cells. PMID- 10847766 TI - Primed and replicating but uncommitted T helper precursor cells show kinetics of differentiation and commitment similar to those of naive T helper cells. PMID- 10847767 TI - New insight into the mechanisms underlying Th2 cell development and susceptibility to Leishmania major in BALB/c mice. PMID- 10847768 TI - Influence of host and parasite factors on the innate immune response and Th2 stability following infection with Leishmania major. PMID- 10847769 TI - Non-MHC-linked genetic polymorphism controls selective development of Th2 but not inhibition of Th1 cells by soluble antigen administration. PMID- 10847770 TI - Loci influencing development of Th responses. Identification from in vitro analysis. PMID- 10847771 TI - Genetic control of T helper subset differentiation in Leishmania major infection of mice. PMID- 10847772 TI - Roles of Nramp1, HLA, and a gene(s) in allelic association with IL-4, in determining T helper subset differentiation. PMID- 10847773 TI - Report on the International Symposium on Vaccinology, November 18-20, Paris, France, organized by the Academie des Sciences and the Marcel Merieux Foundation. PMID- 10847774 TI - [Comment on R. Smektala et al.: Femoral neck fracture: analysis of results of external quality assurance]. PMID- 10847775 TI - [Future oriented clinical research: gene therapy]. PMID- 10847776 TI - [Help for diabetic patients--the discovery of insulin and early hormone research in the Deutsche Medizinische Wochenschrift]. PMID- 10847777 TI - [The problems in the general theory of work activity (on the 275th anniversary of the Russian Academy of Sciences)]. PMID- 10847778 TI - Geriatrics Educational Conference and Exposition. PMID- 10847779 TI - Balance reciprocal translocation t(1;16)(q12;q11) in two generations of a family. PMID- 10847780 TI - Local cytokine production and acute coronary events. PMID- 10847781 TI - Vineberg revisited. Long-term survival more than two decades after direct surgical myocardial revascularization. AB - A 65-year old patient was referred to our institution for a diagnostic catheterization. 23 years before, a direct surgical myocardial procedure using the Vineberg technique was performed. Currently, the angiogram shows patent left and right internal mammary arteries implanted directly into the myocardium and connecting with the native circulation through collaterals. As the native coronary artery tree shows very severe three vessel disease, it is hypothesized that the major contribution of myocardial perfusion comes from the implanted vessels. This is the first case to show a long-term success of the Vineberg operation, with persistence of myocardial perfusion through newly formed vasculature. PMID- 10847782 TI - Atrial flutter resulting from right atrial compression by a hepatic hydatid cyst. PMID- 10847783 TI - The effect of adrenal hormones on the circulation. PMID- 10847784 TI - No apparent myocardial damage following 20,680 joules DC countershocks. PMID- 10847785 TI - A case of Holt-Oram Syndrome (Heart-Hand Syndrome) PMID- 10847786 TI - Pain management: a synopsis. PMID- 10847787 TI - Nomenclature for factors of the HLA system, update January 2000. PMID- 10847788 TI - Regarding Bentel et al. IJROBP 1999;44:1017-1025. PMID- 10847789 TI - Is the prostrate alpha/beta ratio of 1.5 from Brenner & Hall a modeling artifact. PMID- 10847790 TI - Regarding Chan et al. IJROBP 1999;45:265-270. PMID- 10847791 TI - Delayed toxidromes. PMID- 10847792 TI - Mitral valve prolapse versus panic disorder in patients with chest pain. PMID- 10847794 TI - Transverse myelopathy, a rare complication of mixed connective tissue disease: comparison with SLE related transverse myelopathy. PMID- 10847793 TI - No association between the APOE epsilon4 allele and outcome and susceptibility in primary progressive multiple sclerosis. PMID- 10847795 TI - Plasmapheresis in multifocal motor neuropathy: a case report. PMID- 10847796 TI - Ethambutol related headache in HIV infection: sensitisation to normal CSF pressure? A case report. PMID- 10847797 TI - Transcranial Doppler sonography in acute stroke. PMID- 10847798 TI - Giant cell arteritis of the cervical radicular vessels presenting with diaphragmatic weakness. PMID- 10847799 TI - Unidirectional dyslexia in a polyglot. PMID- 10847800 TI - Dyslexia in Hebrew. PMID- 10847801 TI - Kenneth K. Carroll (1923-1998). PMID- 10847802 TI - Zinc and Health: Current Status and Future Directions. Proceedings of a workshop. Bethesda, Maryland, USA. November 4-5, 1998. PMID- 10847803 TI - A piece of my mind. Who cares about Tarawa? PMID- 10847804 TI - From the Centers for Disease Control and Prevention. Serogroup W-135 meningococcal disease among travelers returning from Saudi Arabia--United States, 2000. PMID- 10847805 TI - From the Centers for Disease Control and Prevention. Surveillance for adverse effects associated with anthrax vaccination--US Department of Defense, 1998-2000. PMID- 10847806 TI - From the Centers for Disease Control and Prevention. Prevalence of leisure-time physical activity among overweight adults--United States, 1998. PMID- 10847807 TI - From the Centers for Disease Control and Prevention. Progress toward global poliomyelitis eradication, 1999. PMID- 10847808 TI - JAMA Patient Page. Violence in the media. PMID- 10847809 TI - [Hypoglycemic agents to improve insulin resistance]. PMID- 10847810 TI - [Infectious diseases]. PMID- 10847811 TI - [Guideline for radionuclide detection from sentinel lymph nodes. Working Group for the Preparation of Guidelines for the Internal Use of RI]. PMID- 10847812 TI - [Autochthonous malaria in Guadeloupe: two cases recorded in 1963 and 1965]. PMID- 10847813 TI - [The French Antilles cannot be considered a malaria risk destination]. PMID- 10847814 TI - Brain tumours and mobile phones? PMID- 10847815 TI - Chapter 5. Hospitalizations due to injury: inpatient medical records data. PMID- 10847816 TI - Chapter 7. Defense and Veterans Head Injury Program: a model injury registry. PMID- 10847817 TI - Chapter 9. Conclusions and recommendations of the DoD Injury Surveillance and Prevention Work Group. PMID- 10847818 TI - Chapter 1. Process of injury data acquisition and reporting. PMID- 10847819 TI - Chapter 8. Total Army injury and health outcomes database: a model comprehensive research database. PMID- 10847820 TI - Chapter 2. Deaths due to injuries: casualty office data. PMID- 10847821 TI - Chapter 4. Disabilities related to the musculoskeletal system: Physical Evaluation Board Data. PMID- 10847822 TI - Chapter 3. Fatal and nonfatal accidents/mishaps: safety center data. PMID- 10847823 TI - Chapter 6. Injuries treated in outpatient clinics: surverys and research data. PMID- 10847824 TI - Re: "A new vision of collective bargaining". PMID- 10847825 TI - Re: "Telephone triage and consultation: an emerging role for nurses". PMID- 10847826 TI - Re: "Telephone triage and consultation: an emerging role for nurses". PMID- 10847827 TI - N-acetylcysteine for chronic obstructive pulmonary disease? PMID- 10847828 TI - Do twitchy airways accelerate the bad effects of smoking? PMID- 10847829 TI - A response to 'A rationale for staged teaching of basic life support'. PMID- 10847830 TI - Transeosophageal echocardiograph assessment of mitral valve position during prolonged cardiopulmonary resuscitation and in moderate hypothermia. PMID- 10847831 TI - [Planning of a continuing education program in community health for the elderly]. PMID- 10847832 TI - Fate and transport of agricultural chemicals in the Mississippi River Basin. Proceedings of a conference. Charleston, South Carolina, USA. 8-12 March 1999. PMID- 10847833 TI - Reversibility of HIV drug resistance. PMID- 10847834 TI - Dioxin draft sparks controversy. PMID- 10847835 TI - New clues to how the TB bacillus persists. PMID- 10847836 TI - Biomedical ethics. HHS plans to overhaul clinical research rules. PMID- 10847837 TI - Air pollution. Three Asian nations launch joint study. PMID- 10847838 TI - Hypertension. A DASH of data in the salt debate. PMID- 10847839 TI - Virology. The boom in biosafety labs. PMID- 10847841 TI - European science. A blow to Austria's scientific revival. PMID- 10847840 TI - Public health. Aluminum is put on trial as a vaccine booster. PMID- 10847842 TI - NAS report under scrutiny. PMID- 10847843 TI - Snake origins. PMID- 10847845 TI - Immortality, anyone? PMID- 10847844 TI - Immortality, anyone? PMID- 10847846 TI - Immortality, anyone? PMID- 10847847 TI - Report clarification. PMID- 10847848 TI - Profile: Donald Kennedy--the next Editor-in-Chief of Science. PMID- 10847849 TI - Ecology. Receding forest edges and vanishing reserves. PMID- 10847850 TI - Structural biology. A chloride pump at atomic resolution. PMID- 10847851 TI - Interfering with gene expression. PMID- 10847852 TI - Matching the transcription machinery to the right DNA. PMID- 10847853 TI - Transposable elements: friends, foes, or merely fellow travelers? PMID- 10847854 TI - Cell biology for a new millennium. PMID- 10847855 TI - [The conference dedicated to the 70th anniversary of the Department of Medical Service Organization and Tactics of the Military Medical Academy]. PMID- 10847856 TI - [Dissertations defended at the end of 1999]. PMID- 10847857 TI - Leipzig 2000. Abstracts of the International Symposium and 45th Meeting of the German Society for Neuropathology and Neuroanatomy, 29 March-1 April 2000, Leipzig, Germany. PMID- 10847858 TI - Proceedings of the 3rd Tannin Conference. In honor of Professor Edwin Haslam. Bend, Oregon, USA. July 20-25, 1998. PMID- 10847859 TI - Catechin and procyanidin levels in French wines: contribution to dietary intake. PMID- 10847860 TI - Beverly Malone on healthcare quality, policy, and nursing. Interview by Mary Lou Brunell and Jacqueline F. Byers. PMID- 10847861 TI - Root cause analysis in response to a "near miss". AB - Root cause analysis is a technique used in determining the real cause of an event. The Sentinel Event Policy of the Joint Commission on Accreditation of Healthcare Organizations requires a root cause analysis of a sentinel event but not a "near miss." However, root cause analysis is an important process for preventing another near miss from the same or similar cause. This article reviews the background of basic root cause analysis and provides an example of the process. Changes in policy and procedures for patient-controlled analgesia therapy, as a result of the root cause analysis, are described. PMID- 10847862 TI - Testing the reliability and validity of a measure of safety climate. AB - The lack of compliance with universal precautions (UP) is well documented across a wide variety of healthcare professions and has been reported both before and after the enactment of the Occupational Safety and Health Administration's Bloodborne Pathogens Standard. Gershon, Karkashian, and Felknor (1994) found that several factors correlated significantly with healthcare workers' lack of compliance with UP, including a measure of organizational safety climate (e.g., the employees' perception of their organizational culture and practices regarding safety). We conducted a secondary analysis using data from a cross-sectional survey of a convenience sample of 1,746 healthcare workers at risk of occupational exposure to bloodborne pathogens to assess the validity and reliability of Gershon's measure of safety climate. Findings revealed no relationship between safety climate and employees' gender, age, education, tenure in position, profession, hours worked per day, perceived risk, attitude toward risk, and training. An association was demonstrated between safety climate and (1) healthcare worker compliance with UP and (2) the availability of personal protective equipment, providing support for the construct validity of this measure of safety climate. These findings could be used by occupational health professionals to assess employees' perceptions of the safety culture and practices in the workplace and to guide the institution's risk management efforts in association with U.P. PMID- 10847863 TI - Computerized interdisciplinary assessment. AB - Systems and technology that support the collection and dissemination of healthcare information improve service levels, enhance quality, and improve continuity of care. The delivery of patient care is changing rapidly because sophisticated clinical information systems are providing fast, organized access to large amounts of patient information (Ornstein et al., 1997). The Carl T. Hayden Veterans Affairs Medical Center in Phoenix developed and implemented a computerized process for interdisciplinary assessment and treatment of patients. This was accomplished within the framework of a recent reorganization into service lines and the initiation of a computerized medical record system. Traditional frameworks for change management often include a methodical picture of the required steps or phases of the change process. The rapid rate of change, both technologically and in the healthcare system, however, defies the application of some aspects of traditional change theory. A model for change that considers realistic change strategies, including good planning and identification of barriers, was used to facilitate the move to computerized interdisciplinary assessment (Daly, Button, Prophet, Clarke, & Androwich, 1997). PMID- 10847864 TI - HHS secretary and Surgeon General launch Healthy People 2010. PMID- 10847865 TI - Continuous quality improvement efforts decrease postoperative ileus rates. AB - This article describes a continuous quality improvement team's efforts to reduce the number of patients at risk for developing a postoperative ileus. This study was limited to patients who underwent a surgical procedure that did not involve the bowel. Patients who developed an ileus after the surgical procedure (study group) were compared with patients who did not develop an ileus after the same procedure (control group). Principal factors associated with a postoperative ileus included a low serum albumin level, excessive use of narcotics, and a history of a previous gastrointestinal condition. Practice changes were made as a result of these findings. A follow-up chart review revealed a decrease in the postoperative ileus rate. PMID- 10847867 TI - Reducing errors in medicine. PMID- 10847866 TI - Developing a clinical violence prevention and intervention plan for psychiatric mental healthcare settings. AB - Each year thousands of patients and staff are assaulted in psychiatric mental health settings. Because there are many reasons for such assaults, employers should develop a comprehensive approach to focus on this issue. A proposed format for a clinical violence prevention and intervention plan is presented as a clinical management tool to address patient assaults in psychiatric mental health settings, along with the rationale, use, and components of such a plan. PMID- 10847868 TI - Learning from quality systems in different countries: quality improvement across borders? PMID- 10847869 TI - Assessing good quality dental care. PMID- 10847870 TI - The quality of general dental care: public and users' perceptions. AB - BACKGROUND: Systematic evidence about how the public and users perceive and experience the quality of general dental care is in short supply, particularly in light of the recent changes in the general dental service. The study reported here attempted to fill this gap. OBJECTIVES: To identify the criteria the public and users adopt in evaluating the quality of general dental care, and to identify the extent and nature of perceived concerns with general dental care. DESIGN: Postal questionnaires were sent to random samples of adults living in an inner city area (I) and semi-rural area (R) in southern England. Fifty six per cent (1499) in area R and 48% (1388) in area I completed the questionnaire after four mailings. Follow up face-to-face interviews were done with a purposive subsample (n = 50) of responders from the postal survey. MAIN MEASURES: Public/user views about quality of dental care were measured through groups of questions about the importance of and satisfaction with different aspects of dental care (access/availability including cost; facilities; technical skills; and interpersonal care) and a scale (Dentsat) measuring general satisfaction was constructed from questions on different aspects of care. RESULTS: Evaluation of quality of general dental care from the users' point of view hinges on perceived technical skills, particularly pain management. Major dissatisfaction stems from concerns about costs of dental care and privatisation. CONCLUSIONS: The criteria adopted by the public/users to assess general dental care are similar to other areas of health care, apart from the priority placed on technical skills and pain management. However, the major source of decline in satisfaction with the quality of general dental care is the barrier to access created by the rising cost of dental care and the increasing involvement of dentists in private practice. This evidence suggests that the public and users find the drift towards private practice and away from NHS practice a major source of concern. PMID- 10847871 TI - Measuring patients' attitudes to care across the primary/secondary interface: the development of the patient career diary. AB - BACKGROUND: A growing number of new ways of organising services across the primary/secondary interface are being introduced and evaluated. The principal motive for such reorganisation is to improve the efficiency of health care. However, unless the impact of the new services on patients is investigated and taken into account, it is possible that patients' reactions could be negative, a factor that could lead to unexpected consequences in the use and costs of services. OBJECTIVE: To develop a measure of patients' attitudes towards care across the interface between primary and secondary care. DESIGN: Generation of questions to be included in the measure from a qualitative study of patients' experiences of care across the interface; administration of pilot versions of the measure to samples of patients referred to secondary care; refinement of questions guided by analysis of response patterns, principal components analysis and internal consistency; administration of the final version of the patient career diary in complete form retrospectively to patients referred to secondary care, and one section alone to patients attending outpatient departments for follow up appointments. Face validity was assessed by analysis of open comments in a sample of 50 diaries, and review of the diary by 34 health professionals. Construct validity was assessed by investigation of levels of correlation between components of each section of the diary and the components of the healthcare section overall. SETTING: In the final field test, patients were attending various hospital services, including cardiology, dermatology, neurology, gynaecology, general surgery, general medicine, ophthalmology, trauma and orthopaedics, and gastroenterology. RESULTS: The final version of the diary included 109 questions in seven sections: general practitioner (GP) visits and referral, other GP visits, first outpatient visit, other outpatient visits, inpatient stay and discharge, care after discharge, and care overall. Response rates were poor for retrospective completion of the entire diary, but excellent when a section was given separately. Principal components analysis confirmed that components relating to issues identified as important to patients in the qualitative study had been included in the diary. Levels of internal consistency were good, and comments of patients and health professionals supported validity. CONCLUSION: The patient career diary is a valid and reliable measure of patients' attitudes to care across the interface. It should be given in sections to ensure adequate response rates, and is suitable for use in the evaluation or quality of patterns of care across the interface. In future, the impact on patients of new ways of organising services across the interface should be investigated by use of measures such as the patient career diary. PMID- 10847872 TI - Practice visits as a tool in quality improvement: mutual visits and feedback by peers compared with visits and feedback by non-physician observers. AB - OBJECTIVE: To evaluate and compare the effects of two programmes of assessment of practice management in a practice visit: mutual visits and feedback by peers compared with visits and feedback by non-physician observers. DESIGN: Prospective, randomised intervention study, with follow up after one year. SETTING: General practices in the Netherlands in 1993 and 1994. SUBJECTS: A total of 90 general practitioners (GPs) in 68 practices; follow up after one year comprised 81 GPs in 62 practices. MAIN MEASURES: Scores on indicators and dimensions of practice management in the visit instrument to assess practice management and organisation (a validated Dutch method to assess practice management in a practice visit). Change was defined as the difference in score between the first visit and the visit after one year on 208 indicators and on 33 dimensions of practice management. RESULTS: Data of 44 mutual visits by peers were compared with data of 46 visits by non-physician observers. After a year both programmes showed improvements on many aspects of practice management, but different aspects changed in each of the two programmes. After mutual practice visits, GPs scored significantly higher on content of the doctor's bag, on collaboration with colleagues, on collaboration with other care providers, and on accessibility of patient information than after a visit by a non-physician observer. The visits by non-physician observers resulted in a higher score on extent of use of records and on assessment on outcome and year report. CONCLUSION: Change after mutual practice visits and feedback by peers is more marked than after a visit and feedback by a non-physician observer. PMID- 10847873 TI - Practice visits as a tool in quality improvement: acceptance and feasibility. AB - OBJECTIVE: To evaluate the feasibility and acceptance of (a) two programmes of assessment of practice management in a practice visit: mutual practice visits and feedback by peers versus visits and feedback by non-physician observers and (2) the practice visit method used in these programmes (the visit instrument to assess practice management and organisation (VIP)--a validated Dutch tool). DESIGN: Prospective, randomised intervention study with the two programmes, follow up after one year. General practitioners (GPs) were visited after each programme and after the revisits by non-physician observers a year later. SETTING: General practices in the Netherlands in 1993 and 1994. SUBJECTS: A total of 90 GPs in 68 practices. At follow up after 1 year there were 81 GPs in 62 practices. MAIN MEASURES: Scores (mainly five point scales) for questions on appreciation and acceptance; after the follow up visit a year later, scores for questions on feasibility and practicality of the improved procedure and feedback report. RESULTS: Data of 44 mutual visits by peers were compared with data of 46 visits by non-physician observers. A visit by a non-physician observer was appreciated significantly more. After the practice visit at one year follow up, the participants reported to have appreciated the visit and the feedback report and to prefer feedback of a non-physician observer to that of a peer. Participants' reports on the procedure and the presentation of the feedback provided clues for the improvement of visit procedures. CONCLUSIONS: A practice visit and feedback by a non-physician observer is more appreciated than a visit and feedback by a colleague. A practice visit with the VIP by a non-physician observer is a simple, easy, and well accepted method for assessing practice management. PMID- 10847874 TI - Judging journalism: how should the quality of news reporting about clinical interventions be assessed and improved? PMID- 10847875 TI - A framework for effective management of change in clinical practice: dissemination and implementation of clinical practice guidelines. AB - Theories from social and behavioural science can make an important contribution to the process of developing a conceptual framework for improving use of clinical practice guidelines and clinician performance. A conceptual framework for guideline dissemination and implementation is presented which draws on relevant concepts from diffusion of innovation theory, the transtheoretical model of behaviour change, health education theory, social influence theory, and social ecology, as well as evidence from systematic literature reviews on the effectiveness of various behaviour change strategies. The framework emphasises the need for preimplementation assessment of (a) readiness of clinicians to adopt guidelines into practice, (b) barriers to change as experienced by clinicians, and (c) the level at which interventions should be targeted. It also incorporates the need for multifaceted interventions, identifies the type of barriers which will be addressed by each strategy, and develops the concept of progression through stages of guideline adoption by clinicians, with the use of appropriately targeted support strategies. The potential value of the model is that it may enable those involved in the process of guideline dissemination and implementation to direct strategies to target groups more effectively. Clearly, the effectiveness and utility of the model in facilitating guideline dissemination and implementation requires validation by further empirical research. Until such research is available, it provides a theoretical framework that may assist in the selection of appropriate guideline dissemination and implementation strategies. PMID- 10847877 TI - Improvement through inspection? The development of the new Commission for Health Improvement in England and Wales. PMID- 10847876 TI - Clinical governance: bridging the gap between managerial and clinical approaches to quality of care. AB - Clinical governance has been introduced as a new approach to quality improvement in the UK national health service. This article maps clinical governance against a discussion of the four main approaches to measuring and improving quality of care: quality assessment, quality assurance, clinical audit, and quality improvement (including continuous quality improvement). Quality assessment underpins each approach. Whereas clinical audit has, in general, been professionally led, managers have driven quality improvement initiatives. Quality assurance approaches have been perceived to be externally driven by managers or to involve professional inspection. It is discussed how clinical governance seeks to bridge these approaches. Clinical governance allows clinicians in the UK to lead a comprehensive strategy to improve quality within provider organisations, although with an expectation of greatly increased external accountability. Clinical governance aims to bring together managerial, organisational, and clinical approaches to improving quality of care. If successful, it will define a new type of professionalism for the next century. Failure by the professions to seize the opportunity is likely to result in increasingly detailed external control of clinical activity in the UK, as has occurred in some other countries. PMID- 10847879 TI - Patient care: what drives us to change? PMID- 10847880 TI - The learning organisation: a necessary setting for improving care? PMID- 10847881 TI - The increasing importance of patient surveys. PMID- 10847878 TI - What type of filling? Best practice in dental restorations. PMID- 10847882 TI - Perceptions of good medical practice in the NHS: a survey of senior health professionals. AB - OBJECTIVES: To categorize senior health professionals' experience with poor medical practice in hospitals and in general practice, to describe perceptions which senior NHS staff have of good medical practice, and to describe how problems of poor medical practice are currently managed. DESIGN: A postal questionnaire survey. The questionnaire sought perceptions of good medical practice, asked participants to characterise deviations from good practice, and to describe experience with managing poor performance at the time of the introduction of the General Medical Council (GMC) performance procedures. SETTING: A range of NHS settings in the UK: hospital trusts, health authorities/boards, local medical committees, community health councils. SUBJECTS: Senior health professionals involved in the management of medical professional performance. MAIN MEASURES: Perceptions of what constitutes good medical practice. RESULTS: Most respondents considered that persistent problems related to clinical practice (diagnosis, management, and outcome and prescribing) would require local management and, possibly, referral to the GMC performance procedures. Informal mechanisms, including informal discussion, education, training, and work shifting, were the most usual means of handling a doctor whose performance was poor. Many took a less serious view of deficiencies in performance on manner and attitude and communication, although consultation skills rather than technical skills comprised the greatest number of complaints about doctors. CONCLUSIONS: Senior NHS professionals seem reluctant to consider persistently poor consultation skills in the same critical light as they do persistently poor technical practice. These attitudes may need to change with the implementation of clinical governance and updated guidance from the GMC on what constitutes good medical practice. PMID- 10847884 TI - Measurement of patient perceptions of pain and disability in relation to total hip replacement: the place of the Oxford hip score in mixed methods. AB - OBJECTIVES: To describe the practical difficulties experienced by patients when completing the Oxford hip score, and to highlight the need to reconsider aspects of its structure and conceptual base. DESIGN: Qualitative study incorporating the Oxford hip score in semi-structured interviews with patients before and four months after their operation. SETTING: Two hospitals in the North of England. SUBJECTS: Osteoarthritic patients undergoing primary elective total hip replacement. RESULTS: Use of the Oxford hip score provided quantitative data on disability in the sample, particularly about pain and immobility. It also facilitated the collection of qualitative data, serving as a useful starting point for interviews and as a prompt for indepth discussion. Concerns about the clarity, coverage, and content validity of the score were identified, however, raising questions about the measure's conceptual base. CONCLUSION: The Oxford hip score was found to be a useful precursor to the semi-structured interviews. However, deficiencies in instruction and lack of clarity in purpose have implications for its ongoing development and future application, both in this type of study and other, more general, contexts. PMID- 10847883 TI - Development and validation of a core outcome measure for palliative care: the palliative care outcome scale. Palliative Care Core Audit Project Advisory Group. AB - OBJECTIVES: To develop an outcome measure for patients with advanced cancer and their families which would cover more than either physical symptoms or quality of life related questions. To validate the measure in various specialist and non specialist palliative care settings throughout the UK. DESIGN: A systematic literature review of measures appropriate for use in palliative care settings was conducted. In conjunction with a multidisciplinary project advisory group, questions were chosen for inclusion into the scale based on whether they measured aspects of physical, psychological, or spiritual domains pertinent to palliative care, and whether similar items had shown to be valid as part of another measure. A staff completed version was developed to facilitate data collection on all patients throughout their care, and a patient completed version was designed to enable the patient to contribute to the assessment of their outcomes when possible. A full validation study was conducted to evaluate construct validity, internal consistency, responsiveness to change over time, and test-retest reliability. Assessments were timed. SETTING: Eight centres in England and Scotland providing palliative care, including inpatient care, outpatient care, day care, home care, and primary care. PATIENTS: A total of 450 patients entered care during the study period. Staff collected data routinely on patients in care long enough to be assessed (n = 337). Of these, 262 were eligible for patient participation; 148 (33%) went on to complete a questionnaire. MAIN MEASURES: The Palliative Care Outcome Scale (POS), the European Organisation for Research on Cancer Treatment, and the Support Team Assessment Schedule. RESULTS: The POS consists of two almost identical measures, one of which is completed by staff, the other by patients. Agreement between staff and patient ratings was found to be acceptable for eight out of 10 items at the first assessment. The measure demonstrated construct validity (Spearman rho = 0.43 to 0.80). Test/re-test reliability was acceptable for seven items. Internal consistency was good (Cronbach's alpha = 0.65 (patients), 0.70 (staff)). Change over time was shown, but did not reach statistical significance. The questionnaire did not take more than 10 minutes to complete by staff or patients. CONCLUSION: The POS has acceptable validity and reliability. It can be used to assess prospectively palliative care for patients with advanced cancer. PMID- 10847885 TI - Is the emergency readmission rate a valid outcome indicator? AB - OBJECTIVES: The principal aim was to determine whether the emergency readmission rate varies between medical specialties, and to identify whether differences in emergency readmission rates between hospital trusts can be reduced by standardising for specialty. Possible factors influencing emergency readmission were also investigated, including frequency of previous admission and cause of readmission. DESIGN: Emergency readmission rates were obtained from the Scottish Morbidity Record scheme (SMR1) using record linkage, standardised for age and sex. Rates throughout Scotland were analysed by specialty, and rates for general medicine compared among teaching hospital trusts. Cause of emergency readmission was determined from hospital records in a random sample (177 patients). SETTING: Medical specialties throughout Scotland. SUBJECTS: All patients readmitted as an emergency within 28 days of discharge (October 1990 to September 1994). RESULTS: Emergency readmissions varied markedly between medical specialties, with highest rates in nephrology (24.2%, 95% CI 23.5 to 24.8) and haematology (20.4%, 95% CI 19.9 to 20.9), and the lowest in homeopathy (2.2%, 95% CI 1.6 to 2.7) and metabolic diseases (3.5%, 95% CI 2.4 to 4.5). The largest number of emergency readmissions was in general medicine, accounting for 63% of the total. Restricting emergency readmission rates to general medicine significantly altered previous rates. In the year preceding the emergency readmission, 59% of all patients had been admitted to hospital at least once, and most emergency readmissions (73.3%) resulted from a chronic underlying condition. CONCLUSIONS: Significant variations in emergency readmission rates occurred between medical specialties, suggesting that differences between hospital trusts are influenced by differences in specialties and thus case mix. The majority of emergency readmissions occurred in patients with an underlying chronic condition, and many had a history of multiple previous hospital admissions. The emergency readmission rate is therefore unlikely to be a valid outcome indicator reflecting quality of care until routine data are available for standardisation by case mix. PMID- 10847886 TI - A team quality improvement sequence for complex problems. AB - To solve complex quality problems teams need to follow a systematic sequence of inquiry and action. In this article a practical description of a team quality improvement sequence (TQIS) is given based on the experience of the more successful teams in the Norwegian total quality management experiment. There are nine phases in the sequence and teams have the flexibility to choose the best quality methods for completing each phase. The strengths of the framework are in ensuring that personnel time is used cost effectively and that changes are made which result in measurable improvement. One limitation is that the framework has not been as widely tested as FOCUS-PDCA (find, organise, clarify, understand, select-plan, do, check, act) and other frameworks to which the TQIS framework is compared. It is proposed that if team projects are to be the main vehicle for quality improvement, then their work must be made more cost effective. The article aims to stimulate research into the conditions necessary for different quality teams to be successful in health care, and draws on the research to propose a "risk of team failure index" to improve the management of such teams. PMID- 10847887 TI - Quality of life as an instrument for need assessment and outcome assessment of health care in chronic patients. PMID- 10847888 TI - Changing clinical practice: views about the management of adult asthma. AB - A case study of clinical practice in adult asthma is presented. The case is part of a larger project, funded by the North Thames NHS Executive Research and Development Programme, that sought to explore the part played by clinicians in the implementation of research and development into practice in two areas: adult asthma and glue ear in children. The first case of glue ear in children was reported in a previous issue of this journal (Quality in Health Care 1999;8:99 107). Background information from secondary sources on the condition, treatment, and organisation and location of care is followed by an account of the results of semistructured interviews with 159 clinicians. The findings are reported in two sections: clinical management and the organisation of care, and clinicians' accounts of what, why, and how they introduce changes into their practice. The way clinicians talk about their learning, their expressed views on acceptable practice, and their willingness to change were shown to be informed by construction of legitimate and sufficient evidence, respected colleagues, and accumulated individual experience. There was little open acknowledgment of the influence of organisational factors in influencing practice. To investigate whether relationships between task performance and organisational arrangements found in other sectors apply to UK health, more robust measures by which performance can be evaluated are needed. PMID- 10847889 TI - Management of sickness absence: a quality improvement study from Slovenia. AB - PROBLEM: A need to improve the communication system between general practitioners (GPs) and the national health insurance institute's (NHII) committee of experts for the referral and approval of sickness leave for patients. DESIGN: A structured low cost quality improvement method for motivating GPs to change their current practice was developed. BACKGROUND AND SETTING: The study was done in Kranj health district in Slovenia. GPs and members of the committee of experts identified potential problems using a cause and effect diagram. The study period for baseline data collection was from November 1996 to December 1996, and the re evaluation took place in May 1997. All GPs in Kranj health district (n = 78) took part. Data were collected on 443 patients referred by GPs to the NHII committee during the first phase of the study and 590 patients during the re-evaluation phase. KEY MEASURES FOR IMPROVEMENT: Reducing the number of cases reported by members of the committee of experts as causing problems after the intervention. Feedback to GPs about the success of the process. STRATEGIES FOR CHANGE: A combination of methods was used: posted feedback, a guideline on record keeping, and a guideline, called AID (analysis of incidental deviations from expected service--in Slovene: analiza izjemnih dogodkov), on processing medical documentation. EFFECTS OF CHANGE: An overall drop was observed in the number of cases that caused problems (from 44% to 26%, p < 0.001). The most common problem at baseline (19.4% of the problems) was the seventh most common at the re evaluation, then contributing only 9.2% of total problems (p = 0.02). LESSONS LEARNT: The results support a quality improvement philosophy that empowers "owners" of the process to be the key resource in managing change, and they show the importance of the inner motivation of those involved. Despite working in a country undergoing transition, medical professionals were still willing to improve their performance. Nevertheless, structures and funding are needed to foster quality improvement initiatives and implement national policy on quality in health care. PMID- 10847891 TI - The battle of pork rind hill. PMID- 10847892 TI - Drugged-out toddlers. A new study documents an alarming increase in behavior altering medication for preschoolers. PMID- 10847893 TI - Decoding the human body. PMID- 10847894 TI - A revolution in medicine. PMID- 10847895 TI - The gold rush. PMID- 10847890 TI - Management of gynaecological cancers. PMID- 10847896 TI - It's not 'all in the genes'. The environment you grow up in is as important as your DNA in determining the person you ultimately become. PMID- 10847897 TI - AIDS crushes a continent. Every minute 11 people worldwide are infected with HIV. Ten are in sub-Saharan Africa. PMID- 10847898 TI - Senator Wayne Allard's crusade: returning power to the people. Interview by Val J. Halamandaris. PMID- 10847899 TI - Health in the balance--from home care to earth care. PMID- 10847900 TI - Demography is destiny: forecasting a bright future for home care and hospice. PMID- 10847901 TI - Working together for the future of home care. PMID- 10847902 TI - Quo vadis home care: charting the course for the future. PMID- 10847903 TI - Faces of caring: a tribute to Deepak Chopra, MD. PMID- 10847904 TI - Strengthening our role as healers of the body and the spirit. PMID- 10847905 TI - Healing the soul: the challenge for home care in the 21st century. PMID- 10847906 TI - Congressman Steven T. Kuykendall's mission: keeping America strong and serving the public interest. Interview by Val J. Halamandaris. PMID- 10847907 TI - A conversation with Mike Vance. PMID- 10847908 TI - The doctor as a hospice patient. PMID- 10847909 TI - Family caregivers key to expansion in private-pay market. PMID- 10847910 TI - 20 reasons home care pays. PMID- 10847911 TI - Dr. Gerald Jampolsky puts forth forgiveness as key to good health. PMID- 10847912 TI - Health information networks: enabling care management in IDSs. AB - To be successful, IDSs require streamlined, information integration across the continuum of care. Having this capability enables IDSs to manage care and costs efficiently, provide physicians with enterprisewide access to eligibility information, and respond to the communication and quality demands of patients. For most IDSs, the quickest and most cost-effective way to achieve overall information-system integration is to implement a health information network (HIN) based on Internet technologies. A HIN enables IDS providers and staff to access requisite administrative and clinical information from throughout the enterprise using a Web browser. PMID- 10847913 TI - Provider terminations: strategies for risk management. AB - Managed care organizations should protect themselves from possible legal action by providers whom they have terminated, whether because of professional incompetence or for reasons related to the organization's business goals. Risk management begins by being aware of state and Federal laws regarding contracts, peer review, restraint of trade, discrimination, and retaliation against a provider. Managed care organizations should adopt a set of criteria for providers' professional competence as well as a list of business objectives that might necessitate the deselection of a provider. Because some courts have upheld a provider's right to a "fair procedure," managed care organizations also should consider offering providers an opportunity to respond to the termination decision. PMID- 10847914 TI - Developing a value-added Web site. AB - Once a healthcare organization has decided to establish a Web site on the Internet, it must next determine its implementation strategy, based on a full understanding of the goals of the site and the range of Web content and service offerings available in the marketplace. Although some organizations may choose to develop and maintain a Web site using exclusively in-house resources, most healthcare organizations will find that they can minimize the costs associated with this effort by making judicious use of outsourcing services. Whichever approach is used, it is important that financial managers charged with allocating resources for Web-site development and maintenance understand the implications, including relative financial impact, of key issues and options. PMID- 10847915 TI - Privacy and security compliance in the E-healthcare marketplace. AB - Complying with security and privacy regulations proposed by HHS in response to the Health Insurance Portability and Accountability Act (HIPAA) will require healthcare managers to address both internal and external business interactions and initiatives. The proposed regulations mandate certain procedures regarding administration, physical safeguards, technical security for data integrity and confidentiality, and technical security against unauthorized access. In particular, the proposed regulations require organizations to contractually ensure that vendors adhere to the regulations. Healthcare organizations also must implement training procedures for staff members who have contact with protected health information and designate a privacy officer to guard against improper disclosure of such information. Documented policies for organizational decision making are vital to an organization's efforts to implement procedures for compliance with the regulations. PMID- 10847916 TI - HIPAA brings new requirements, new opportunities. AB - The passage of the Health Insurance Portability and Accountability Act of 1996 (HIPAA) brought with it the need for Federal rules to implement the act's simplification and cost-reduction efforts. HHS has published proposed rules related to security for the electronic transmission of health information, privacy of individually identifiable health information, transactions and code sets, and national provider and employer identifiers. Additional proposed rules will be published this year for claims attachments and health plan identifiers. Although HIPAA does not require providers to conduct business electronically, the new standards give providers the opportunity to reduce healthcare administrative costs significantly and undertake electronic commerce efficiently and cost effectively. PMID- 10847917 TI - FASB Statement No. 136 clarifies transfers of assets. AB - FASB Statement of Financial Accounting Standards No. 136, Transfers of Assets to a Not-for-Profit Organization or Charitable Trust That Raises or Holds Contributions for Others, provides guidance and establishes accounting standards for the transfer of assets from donors to not-for-profit organizations that may then transfer those same assets to a beneficiary organization. Recipient organizations that accept financial assets from a donor and agree to use those assets on behalf of a specified unaffiliated beneficiary or transfer those assets, the return on investment of those assets, or both to that beneficiary must recognize the assets received from the donor and recognize the assets' fair value as a liability to the beneficiary. The statement describes circumstances in which a transfer of assets to a recipient organization is accounted for as an asset and corresponding liability of the recipient organization, and as an asset and donation revenue by the beneficiary organization because the transfer is irrevocable. PMID- 10847918 TI - Rescuing the hospital-sponsored group practice. AB - Underperforming hospital-sponsored group practices can achieve reasonable financial performance without sacrificing morale among the group's physicians and office staff if six key elements for success can be implemented. These key elements involve formalizing physician leadership and governance, clearly allocating risk, communicating expectations regarding pay for performance, providing meaningful information, keeping overhead expenses under control, and putting an experienced management team in place. Implementing these six elements can help struggling hospital-sponsored group practices achieve a successful turnaround. PMID- 10847919 TI - Antikickback versus stark: what's the difference? PMID- 10847920 TI - The hidden competencies of healthcare: why self-esteem, accountability, and professionalism may affect hospital customer satisfaction scores. AB - Data from 103 for-profit, nonprofit, and government-owned hospitals, spread across about half of the United States clearly show that there are common elements and several core competencies in all hospitals, some probably driven by JCAHO accreditation standards, but others coming from universal experience stemming from the changes in healthcare. The common competencies that are not, in my opinion, driven directly by the JCAHO standards include professionalism, accountability, self-esteem, customer service/focus, communication, information management/using data in decision making, and teamwork. There are several possible connections among the core competencies that suggest that the effects of accountability and possibly self-esteem on such outcomes as patient satisfaction and quality of care should be the subject of more research in healthcare settings. There are, however, several possible interventions to increase the core competency base of any hospital, which can be applied without this research. Executives and managers who attempt to measure and change these common competencies through selection, assessment, organizational system change, or reward and compensation systems will change the competence base of their workforce in critical areas needed in the future healthcare economy. Using a competence model incorporating these competencies may change the culture of the organization toward that which will be needed for survival in the twenty-first century. PMID- 10847921 TI - Risk management and quality improvement: together at last--Part 1. PMID- 10847922 TI - Face-off with death: developing educational programs for dealing with death. PMID- 10847923 TI - Stalking the killer in the corridor--hospitals strive to reduce infection as antibiotic resistance grows. PMID- 10847924 TI - Caregiving from a distance. PMID- 10847925 TI - Disability in older adults: policy implications. PMID- 10847926 TI - Risk management and quality improvement: together at last--Part 2. PMID- 10847927 TI - Nursing unions: the prime time for organizing is now. PMID- 10847928 TI - Distance learning for hospital managers. AB - Telemedicine technology enabled this class to meet. The Chapel Hill instructor could not have traveled to Scotland Neck for the classes, and the class members could not have taken time away from their jobs to travel to Chapel Hill. The technology allowed the participants to fit the classes into their schedules. For the group of managers at this small, isolated hospital, the experience of participating in a management class with an expert was a positive one. They were introduced to standard management practices, learned new skills, and formed a support group/team onsite. The students felt close to the leader, yet the physical distance made her an outsider in a way that encouraged frankness. The technology seemed to foster the best of both worlds--intimacy, yet physical distance and, thus, safety. These new managers were able to take part in a course that taught tangible skills for improving their job performance and, more important, afforded access to resources outside of Halifax County. They were able to step away from their daily routine and interact with outsiders and each other in new ways, without the stress and expense of travel. The results of this pilot study indicate that distance learning is feasible for courses of this kind. Staff burnout and turnover are chronic problems in rural facilities, with isolation contributing to job dissatisfaction. Distance learning offers exciting possibilities for addressing these problems in healthcare settings across the country. PMID- 10847929 TI - Can medicine and chiropractic practice side-by-side? Implications for healthcare delivery. PMID- 10847930 TI - Life-cycle preferences over consumption and health: when is cost-effectiveness analysis equivalent to cost-benefit analysis? AB - This paper studies life-cycle preferences over consumption and health status. We show that cost-effectiveness analysis is consistent with cost-benefit analysis if the lifetime utility function is additive over time, multiplicative in the utility of consumption and the utility of health status, and if the utility of consumption is constant over time. We derive the conditions under which the lifetime utility function takes this form, both under expected utility theory and under rank-dependent utility theory, which is currently the most important nonexpected utility theory. If cost-effectiveness analysis is consistent with cost-benefit analysis, it is possible to derive tractable expressions for the willingness to pay for quality-adjusted life-years (QALYs). The willingness to pay for QALYs depends on wealth, remaining life expectancy, health status, and the possibilities for intertemporal substitution of consumption. PMID- 10847931 TI - Premium subsidies for health insurance: excessive coverage vs. adverse selection. AB - The tax subsidy for employment-related health insurance can lead to excessive coverage and excessive spending on medical care. Yet, the potential also exists for adverse selection to result in the opposite problem-insufficient coverage and underconsumption of medical care. This paper uses the model of Rothschild and Stiglitz (R-S) to show that a simple linear premium subsidy can correct market failure due to adverse selection. The optimal linear subsidy balances welfare losses from excessive coverage against welfare gains from reduced adverse selection. Indeed, a capped premium subsidy may mitigate adverse selection without creating incentives for excessive coverage. PMID- 10847932 TI - Optimal social health insurance with supplementary private insurance. AB - This paper investigates the structure of a National Health Service in which there is compulsory social insurance covering a package of essentials, a given part of individuals' health expenditure, and supplementary private policy topping up the remaining services. The latter insurance contract provides for a co-payment by patients, limiting the so-called "third-party payer" effect. Thus, an individual's health expenditure is divided into three parts: the first covered by social insurance, the second by a private policy and the third out-of-pocket. Such mixed system design has received increasing attention in recent years and has been adopted by several industrialized countries. The conditions for optimal rates of social insurance coverage and of private coinsurance are analysed and discussed. The optimality requirements refer to efficiency as well as equity concerns. PMID- 10847933 TI - Estimating the quality of care in hospitals using instrumental variables. AB - Mortality rates are a widely used measure of hospital quality. A central problem with this measure is selection bias: simply put, severely ill patients may choose high quality hospitals. We control for severity of illness with an instrumental variables (IV) framework using geographic location data. We use IV to examine the quality of pneumonia care in Southern California from 1989 to 1994. We find that the IV quality estimates are markedly different from traditional GLS estimates, and that IV reveals different determinants of quality. Econometric tests suggest that the IV model is appropriately specified, that the GLS model is inconsistent. PMID- 10847934 TI - The complementarity of teen smoking and drinking. AB - Teen drinkers are over twice as likely as abstainers to smoke cigarettes. This empirical study provides evidence of a robust complementarity between these health behaviors by exploiting the "cross-price" effects. The results indicate that the movement away from minimum legal drinking ages of 18 reduced teen smoking participation by 3 to 5%. The corresponding instrumental variable estimates suggest that teen drinking roughly doubles the mean probability of smoking participation. Similarly, higher cigarette taxes and reductions in teen smoking are associated with a lower prevalence of teen drinking. However, the results which rely on cigarette taxes for identification are estimated imprecisely. PMID- 10847935 TI - The price elasticity of opium in Taiwan, 1914-1942. AB - Between 1895 and 1945, the Japanese colonial government virtually eliminated opium use in Taiwan by licensing and treating existing users, prohibiting sales to others, and raising the price. We evaluate these policies using a two-part model to describe the fraction of the population using opium and consumption among users, and the rational addiction model by Becker et al. (1991). We confirm that opium is addictive and find no evidence supporting the rational addiction hypothesis. Demand is price-elastic with estimated short- and long-run demand elasticities of -0.48 and -1.38. These results have implications for control of other addictive substances. PMID- 10847936 TI - The economics of moral hazard revisited. AB - Pauly's (1968) [Pauly, M., 1968. The economics of moral hazard, Comment, American Economic Review 58, 531-537.] analysis of the welfare loss from insurance assumes that medical care consumption is not determined by income, but recent studies suggest it is. This study argues that (1) Pauly's analysis overstates the welfare loss because it includes the effect of income on consumption, (2) the relevant income effect is derived from income transfer from the healthy to the ill that occur when the probability of illness is less than 1, and (3) the welfare loss can be considered the transaction cost of insurance. PMID- 10847937 TI - Preserving the soul of medicine and physicians: a talk with David Whyte. Interview by Richard L. Reece. AB - Richard L. Reece, MD, interviewed David Whyte, a poet, consultant, and author of The Heart Aroused, on December 15, 1999, to discuss preserving the soul of physicians in corporate America. David describes the soul as "a measure of our belonging in the world. When there is little sense of belonging, there is very little sense of soul." In the workplace, he thinks about whether "people have a sense of belonging to the particular work or the organization." He talks about life in the upper world of the workplace and life in the dark subterranean caves where the soul lives. The soul is where people's true creativity and imagination resides ... and by inviting it into the workplace, organizations and employees can become more successful, innovative, and adaptable. In corporate settings, he uses poetry to bring an understanding of the process of change, helping clients to understand individual and organizational creativity to transform the workplace. The poetry can teach and touch those places that the corporate language cannot speak to. PMID- 10847938 TI - Business leadership as a spiritual discipline. AB - What motivates organizational leaders in their search for spirituality? They seek to integrate their inner journey with their day-to-day professional roles. This article describes how a course in spirituality for executives has provided tools to analyze and clarify intentions, avoid the traps of excessive greed and power, and make decisions that are both compassionate and effective. Andre L. Delbecq, DBA, the Thomas J. and Kathleen L. McCarthy Professor at the Leavey School of Business at Santa Clara University in California, offers seminars in spirituality for organizational leadership through the MBA program and the Center for Executive Development. Delbecq is the first to admit his surprise at the number of executives who have repeatedly asked for courses in spirituality. He talks about how his seminars have helped CEOs and other top executives achieve greater effectiveness in leading organizations. PMID- 10847939 TI - Providing health care to human beings trapped in the poverty culture. AB - The culture of poverty impacts everything patients in this socioeconomic group think and do. If what poor patients say does not sit well with the way we think, that doesn't mean they are wrong. Physicians have to adjust their mental model and think in different cultural terms. The author recently completed his thirtieth year of a career dedicated to providing health care to people living in poverty. He shares seven concepts important in building a mental model that will enable physicians to successfully provide health care to this patient population: (1) Poverty is the number one health problem; (2) we see same diseases as everyone else; (3) patients are trapped in the poverty culture; (4) patients' behavior is often manipulative; (5) compliance is a unique challenge; (6) patients have limited resources; and (7) the ultimate contributors to poverty are unwanted adolescent pregnancy and substance abuse. These concepts can help physicians to be more effective in providing health care to patients living in poverty. They can help them understand what is happening, so that their experience might be fulfilling rather than demoralizing. PMID- 10847940 TI - Managed care's Achilles heel: ethical immaturity. AB - How can physician executives determine the prevailing values in the managed care arena? What are the consequences when values statements are ignored during decision-making? These questions can be answered using a process called ethical reasoning, which is different and more productive than making moral judgments, such as "is managed care good or bad?" Failing to include ethical reasoning in executive offices and boardrooms is a form of ethical immaturity. It fuels public suspicion that managed care's goal may be maximizing profit at all costs, as opposed to seeking reasonable profit through provision of dependable and accessible health care services. One outcome of ethical reasoning is rediscovering the basic truth that running one's business on competitive rather than altruistic principles is ethical whenever greater efficiencies and economic growth enlarge the size of the pie for everyone. Reasonable self-interest is a perfectly acceptable reason to act ethically. The time has come for physician executives to develop a basic understanding of pragmatic ethics, and to appreciate the value of adding ethical reasoning to the decision-making process. PMID- 10847941 TI - The consumer and provider: pillars of the new health care system. AB - Managed care has suffered a public backlash, with complaints increasing across the nation from unhappy patients. The physician community despises the current system and is wrestling for control of clinical decision-making. A health care system that is disliked by the public and is despised by the physician community can never succeed. No health care system or reform is possible without willing or even enthusiastic physician participation because only they can control costs, quality of care, and consumer satisfaction. A successful health care system recognizes that only providers can control quality of care and costs--and will create appropriate incentives that allow physicians to do so without losing the public's trust. The author advocates a new system, where consumers choose provider organizations based on disease expertise and purchase insurance through Internet accessible brokers. Provider organizations assume economic risk and have the detailed know-how to treat a specific disease spectrum better and cheaper. Consumers purchase this new "product" in a competitive market and are the principal benefactors of this market-driven, unmanaged care system. PMID- 10847942 TI - The consumer choice model: a humane reconstruction of the U.S. health care system. AB - "Consumer choice," "defined contribution health programs," "voucher systems," and "health marts" are variations on a theme: employees buying their own health care. This new approach to health care purchasing, which is designed to minimize the role of employers, is being proposed by an array of economists and by both Republican and Democratic legislators as the best way to address the nation's health care ills. Although enabling national legislation is unlikely to pass soon, the debate will nevertheless change the face of health care in America. The prospect is reminiscent of the debate over "Clinton Care" in 1993--although legislation was never passed, managed care rapidly came to dominate the U.S. health care system. As this reform takes hold, beneficiaries will make their own health plan selections but will have more responsibility and may bear more cost. Providers will have to adapt to new, customer-driven requirements for performance, accountability, and communications but will also find opportunities in a marketplace that they will have a major role in shaping. Physicians, health plans, and insurers should understand how these proposals will transform their role in health care. PMID- 10847943 TI - Overworked? Lighten your load. AB - Physician executives are suffering from creeping task migration--putting in too many hours for few appreciable results. Controlling this increasing workload has become one of the most important issues in health care, brought on by too few people trying to do too much. The only way to lessen the workload is to take charge, analyze, and act. Bosses won't care about the details. Several suggestions are presented to help physician executives downsize their workload while not reducing their output: (1) Compare your priorities with your boss's; (2) lighten up on the perfectionism; (3) change expectations; (4) look for "orphans" to cut--those projects that nobody is invested in; (5) don't target symbolic events; and (6) use logic to drive change. If you spend the next few months getting rid of the ineffective, inefficient, and redundant, you'll be ahead of the game. This is an ongoing task, and much easier done every three or four months than once a year. PMID- 10847944 TI - Can you take your soul to work? AB - David White in The Heart Aroused: Poetry and the Preservation of Soul in Corporate America explores ways for professionals to take their souls to work, instead of checking them at the door. "We simply spend too much time and have too much psychic and emotional energy invested in the workplace for us to declare it a spiritual desert bereft of life-giving water." Several ideas are presented to help physician executives preserve their souls in an increasingly corporatized U.S. health care system: (1) Figure out what you are meant to do as your life's calling; (2) know what you think and want; (3) share some of what you think at work, while being careful to not lose your job unless you choose to; (4) be a trustworthy listener and find one; (5) get yourself outside; (6) pay attention to your physical space; and (7) develop some new hobbies or refresh old ones. "One of the disciplines of building a rich soul life seems to be the simple act, on a daily basis, of remembering what is most important to us." PMID- 10847945 TI - Spirituality and the physician executive. AB - The "s" word can now be spoken without flinching in health care organizations. Spirituality is becoming a common topic in management conferences around the world. Many U.S. corporations are recognizing the role of spirituality in creating a new humanistic capitalism that manages beyond the bottom line. Spirituality refers to a broad set of principles that transcend all religions. It is the relationship between yourself and something larger, such as the good of your patient or the welfare of the community. Spirituality means being in right relationship to all that is and understanding the mutual interdependence of all living beings. Physician executives should be primary proponents of spirituality in their organizations by: Modeling the power of spirituality in their own lives; integrating spiritual methodologies into clinical practice; fostering an integrative approach to patient care; encouraging the organization to tithe its profits for unmet community health needs; supporting collaborative efforts to improve the health of the community; and creating healing environments. PMID- 10847947 TI - Human perception of digital displays. PMID- 10847946 TI - Addressing medical errors: the key to a safer health care system. AB - A recent report on patient safety by the Institute of Medicine's Committee on Quality of Health Care in America noted that there are at least 44,000 patient deaths from medical errors each year, placing them as the eighth leading cause of death in the United States. They occur in every aspect of the practice of medicine. Some result in adverse events that harm patients. Can an organized effort to reduce medical errors be effective? Other complex industries have been successful in reducing errors and improving quality. The IOM report argues that the medical community must do the same to ensure a higher quality of care. Both the Clinton administration and Congress have expressed concerns about the frequency of medical errors, as has organized medicine. These findings raise significant policy questions for physician executives' charges with ensuring patient safety. PMID- 10847948 TI - Hospital-wide PACS need tighter data security. PMID- 10847949 TI - Internet challenges PACS as best way to distribute images. PMID- 10847950 TI - Voice recognition saves time, money, but rads resist it. PMID- 10847951 TI - Feds scrutinize supervision of studies done at imaging centers. PMID- 10847952 TI - Perfusion education in the United States at the turn of the century. AB - The challenges facing institutions charged with the delivery of health care have also affected the delivery of education in the health sciences. In the 30 years since the establishment of a process for formalized perfusion education, significant changes have shaped how the fundamentals of perfusion sciences are learned. The establishment and maintenance of a profession can only be secured through the creation of standards that guide educational facilities in the delivery of formalized instruction in a discipline. Perfusion education programs continue to meet these standards but are doing so at a time where resources continue to dwindle and quantitative assessment of manpower issues are fuzzy, at best. The peak of perfusion education programs occurred in 1994, and 5 years later only 25 programs were still accredited by the Committee on Accreditation of Allied Health Programs (CAAHEP). In the past decade, only one new education program has sought accreditation from CAAHEP. There were 41% fewer graduates in the United States in 1998 than in 1992, which follows a 6-year trend in declining numbers of individuals entering the field of perfusion. Individual programs are challenged by the reordering of university and community hospital structures, which often results in critical reviews of resource allocation to perfusion programs. The health of the perfusion profession remains deeply tied to the success of perfusion education programs. Likewise, the health of these programs can only be assured by means of continued solicitation of support and guidance from practitioners who serve as stewards of perfusion technology. PMID- 10847953 TI - Differential expression of inducible nitric oxide synthase in septic shock. AB - During cardiopulmonary bypass (CPB), the septic patient has markedly decreased peripheral vascular resistance as a consequence of endotoxin release from microorganisms. This decrease in peripheral vascular resistance is the result of endotoxin-induced nitric oxide (NO) produced by inducible nitric oxide synthase (iNOS) and endothelial nitric oxide synthase (eNOS). iNOS and eNOS are responsible for the synthesis of NO because of various stimuli, including the bacterial endotoxin, lipopolysaccharide (LPS). We tested the hypothesis that a differential expression of iNOS among human endothelial cells and murine macrophage is dependent upon exposure to endotoxin and various pro-inflammatory cytokines. Using a human endothelial cell line, ECV-304 and murine macrophage cell line, RAW 264.7, we quantified the expression of iNOS with specific FITC conjugated antibodies using fluorescence activated cell sorter (FACS) and NO production with a Bioxytech nitric oxide spectrophotometric assay. This in vitro septic model utilized LPS supported with species-specific interferon-gamma, interleukin-1 beta, and tumor necrosis factor-alpha. The cell type were stimulated for 8 hours with combinations of the cytokines mentioned. The FACS data demonstrated a significant stimulus-dependent increase in iNOS expression among the macrophage groups; however, the stimulated endothelial cells showed no significant change in iNOS expression. The nitric oxide production data demonstrated significant increases in NO production among macrophage stimulated groups; whereas, endothelial stimulated groups exhibit no significant change. We conclude that NO secreted during septic shock is the result of activated macrophage, not the endothelium. The clinical relevance is that the more severe the infectious process, the lower the PVR may be during CPB because of increased NO production from activated macrophage. PMID- 10847954 TI - Investigations into the sterility of manually assembled extracorporeal circuits with vented reservoirs. AB - This study was designed to investigate the ability of an extracorporeal circuit (ECC) with a vented hard shell reservoir to remain sterile for a period of 72 h under dry conditions. The study was conducted in three phases. In Phase One: Two previously published methods for detecting contamination of the ECC were compared. A group of positive controls was collected by contaminating identical circuits with a known level of Enterobacter cloacae (ATTC: 13047) before initiating a regimen of "sample-dilute-sample" culturing. Negative controls for this phase were conducted by randomly sampling 1 L per manufacturer's lot of lactated ringers with each detection method. Culture results suggest that large volume filtration, but not small aliquot sampling, is sensitive to extremely low levels of contamination. No growth was detected in any negative control samples. In Phase Two: 19 ECC consisting of a membrane oxygenator, vented hardshell reservoir, arterial filter, and PVC tubing were removed from their sterile packages, assembled, and left unprotected in the moderate traffic environment of a research laboratory. The circuits were then primed with Lactated Ringer's solution. The prime solution was sampled for aerobic contamination by large volume filtration. None of the 19 samples detected contamination. In Phase Three: 43 ECC identical to the Phase Two circuits were assembled and left unprotected in the substerile pump room. The circuits were then primed, circulated, and cultured as in Phase Two. One of the 43 samples was discarded because of a recognized break in aseptic technique during sample collection. None of the remaining samples detected contamination. Mathematical calculations of binomial probabilities suggest that the chance of an open ECC developing a detectable level of contamination within 72 h of its dry assembly is insignificant. PMID- 10847955 TI - Activated clotting time (ACT) testing: analysis of reproducibility. AB - Activated Clotting Time (ACT) has been the standard for monitoring heparin anticoagulation in cardiac surgery for three decades. Although a 10% coefficient of variation (CV) is the referenced standard for the test, no recent reports of precision are available. The precision of Hemochron FTCA510 (celite) and KACT (kaolin) ACT test tubes was evaluated using a retrospective analysis of results from both laboratory studies and routine clinical usage. Laboratory studies of reproducibility included analysis of the CV from repetitive testing using multiple lots of ACTs. Substrates used included 40 consecutive lots of control plasma and freshly heparinized donor blood. Across the lots of control plasma, the celite ACT yielded an average CV of 5.4% for the normal control level and 4.0% in the abnormal control level (range 3.6-9.7% and 2.7-6.3%, respectively). The KACT showed similar performance for the normal (mean = 4.5%, range 2.2-7.8%) and abnormal (mean = 3.8%, range 2.0-10.0%). These values, significantly less than 10%, reflect the combined variability of both the ACT tests and the lyophilized, single use vial, control material. Fresh whole blood samples exhibited improved ACT precision when compared to this artificial substrate. CVs for the celite ACT range from 0.6-6.0% at one unit heparin/ml blood to 2.4-11.6% at 5 units/ml where clotting times exceed 650 sec. The KACT showed even lower CVs at all heparin levels, with values of 2.4-7.0%. Clinical evaluations included samples (N = 56) collected from cardiac surgery patients with celite ACT values ranging to 744 sec. Duplicate values differed by an average of 7.5 sec or 1.8%. There was only one clinically significant difference in paired values; a 376 sec paired with a 406 sec, 400 sec being the clinical target time. This retrospective data analysis demonstrates that Hemochron ACT variability is significantly less than 10%. PMID- 10847956 TI - Quantitative evaluation of heparin-coated versus non-heparin-coated bypass circuits during cardiopulmonary bypass. AB - The extracorporealization of blood activates various elements of the fibrinolytic, coagulation, and complement systems. It is theorized that advancements in biocompatibility ameliorate many of the changes leading to improved patient management. The purpose of this study was to determine if heparin-coated circuit (HCC) utilization during cardiopulmonary bypass enhances patient outcomes in a cost-effective manner. A search of the English medical literature was completed to identify all clinical, prospective, randomized trials comparing HCC and non-HCC in patients undergoing coronary artery bypass grafting or valvular surgery. Twenty-six papers consisting of a sample size of 1515 patients were identified and included in the study parameters. The study distinguished between Duraflo II and Carmeda coating techniques and matched papers with different heparin loading doses, as well as use of a heparin-coated cardiotomy. Study parameters were matched for all papers and analyzed according to the availability of data. Statistically significant benefits of HCC were found in postoperative blood loss, time in the ICU, end bypass C3a, time to extubation, end bypass lactoferrin, and end platelet count, but not with respect to postoperative chest tube drainage, red blood cell transfusions, and end bypass TAT complex, D-dimers, and BTG. Data comparing the use of coated or uncoated cardiotomy utilization failed to demonstrate a benefit to heparin coating. Several immunological variables were ameliorated when Carmeda HCC was utilized, although data were insufficient to establish a cost-benefit analysis. In conclusion, heparin-coated circuitry provided statistically better results when compared to noncoated circuitry. PMID- 10847957 TI - Heparin-bonded circuit with low systemic anticoagulation in a patient with heparin-induced thrombocytopenia: a case report. AB - Heparin-induced thrombocytopenia (HIT) in patients undergoing open heart surgery has been reported with increasing frequency. Several strategies have been suggested to approach this difficult problem. However, the syndrome is still associated with significant morbidity and mortality. We describe an 82-year-old male with HIT who underwent coronary artery bypass grafting utilizing a heparin bonded cardiopulmonary bypass circuit with very low systemic anticoagulation. Only one unit of packed red blood cells was transfused. The patient recovered uneventfully. This strategy is safe and effective, and, therefore, should be considered in patients with HIT. PMID- 10847958 TI - Avoiding hyperoxemia at the start of cardiopulmonary bypass while optimizing gas flow and temperature. AB - There seems to be a wide range of practice in relation to the optimum oxygen setting before, and at the start of, cardiopulmonary bypass. Even manufacturers of blood oxygenators vary in their suggestions for this phase of extracorporeal circulation. Most of these suggestions are based on peak performance, Association for the Advancement of Medical Instrumentation (AAMI) standards, experience, and legal considerations. Therefore, suggested gas:blood flow ratios will vary from no gas flow at the start of bypass, to a ratio setting of 1:1. On the other hand, suggested inspired oxygen concentrations will generally vary between 0.80 to 1.0 at the start of cardiopulmonary bypass. In regard to perfusate temperatures before going on bypass, there are no clearly defined standards other than those of clinical preference. The manufacturer of the oxygenator used in this study clearly states in the operating instructions that gas flow should be proportional to blood flow at the start of bypass, and gas flow should be turned off when there is no fluid flow through the oxygenator. The presence of hyperoxic perfusates and wide patient/perfusate temperature gradients at the start of bypass has been suspected in the appearance of gaseous microemboli during this critical period. Hyperoxemia during the bypass period is also implicated in the introduction of oxygen free radicals and nitric oxide into the hypoxic myocardium during cardioplegia delivery. Presented here are the results of a randomized clinical study involving 39 adult patients undergoing cardiopulmonary bypass for the surgical treatment of coronary artery disease. All patients were randomly selected into five groupings. The first group had 1 L of gas flow through the perfusate before bypass, and bypass was then started with an FIO2 of 0.80. The second two groups had no gas flow through the perfusate prior to bypass and a starting FIO2 of 0.21. Groups 4 and 5 had 1 L of gas flowing through the perfusate and a starting FIO2 of 0.21. Results indicate that gas flow through Normosol R/Albumin perfusates will prevent the acidosis that is found in this solution when the system is previously flushed with carbon dioxide. Also, suggested high FIO2 settings will produce hyperoxic perfusates at the start of cardiopulmonary bypass. However, the use of an FIO2 of 0.21 at the start of bypass will produce normoxemic conditions that are both safe and reliable for the conduct of initiating cardiopulmonary bypass. PMID- 10847959 TI - A ten-year clinical follow-up study of prosthetic rehabilitation of the edentulous lower jaw with endosteal dental implants. AB - This paper reports on 10 years of clinical experience with implant prosthodontic treatment of total edentulism. During this period, of the 735 implants that were inserted, only 19 were lost, a 97.4% success rate. The authors believe that evaluating the success of implant treatment requires examining the health of the peripilastrium with periodontologic methods. The indices the authors used for such an evaluation were plaque accumulation, crevicular fluid flow rate, and sulcus bleeding; crevicular fluid flow proved the most satisfactory for estimating the health of peri-implant tissues. The results demonstrate that peri implant-tissue health is similar to that of the tissues surrounding nontreated natural teeth and better than that of the tissues surrounding natural teeth wearing crowns. PMID- 10847960 TI - Implant-prosthetic rehabilitation after segmental mandibulectomy and bone grafting. AB - A free vascularized fibula (6 cases) and iliac bone (11 cases) graft were used to reconstruct a mandibular discontinuity defect resulting from cancer surgery in 17 patients. Fifty-two osseointegrated DenTi implants were secondarily placed into the grafted bone, and all the patients underwent prosthetic rehabilitation. (DenTi is an endosseous dental implant system invented and manufactured by Denti System Ltd., Szentes, Hungary.) To date, the maximum follow-up time after prosthetic treatment is 5 years, the minimum is 2 years. During this time, only one of the 52 inserted implants was lost, and that occurred a few days after insertion. Clinical signs of peri-implantitis were observed in only 2 of the fixed prosthetic restorations. Long-term maintenance is essential if such patients are to achieve successful prosthetic restorations and, thus, enhanced lives. PMID- 10847961 TI - Horseshoe Le Fort I osteotomy: an augmentation technique for the severely atrophied maxilla--an eight-year follow-up. AB - Twenty-three patients (19 female, 4 male; 37-69 years old; mean age, 52 years) underwent horseshoe Le Fort I osteotomy from 1989 to 1997; 178 iMZ implants were placed either simultaneously by horseshoe Le Fort I osteotomy (76 implants) or in a second operation 6-12 months later (102 implants). Removable overdentures were made. Twenty-two patients wear an implant-borne overdenture; 1 patient refused an additional operation for placing implants and received a conventional, mucosally supported, upper-jaw prosthesis. The anteroposterior relationship satisfactorily improved in all patients, allowing for class I occlusion. All patients were satisfied with the aesthetic results, denture retention, speech, and mastication. Nevertheless, we lost 8 implants in 1 patient. We placed 178 endosteal implants in 22 patients; 8 were lost in 1 patient, and 7 more implants have been lost in 5 patients. PMID- 10847962 TI - Histomorphologic findings on human bone samples six months after bone augmentation of the maxillary sinus with Algipore. AB - Sinus grafting, a popular and standard treatment for maxillary atrophy, uses a variety of grafting materials. In this study, specimens obtained 6 months after sinus grafting with Algipore were evaluated under light microscopy and showed osseoformation, xenograft degradation, and bone ingrowth into particles. Osteoblastic cells were embedded in the intracorpuscular bone matrix, which indicated that xenograft particles are an osseoconductive scaffold and stimulate matrix deposition. Acute inflammatory responses after insertion of Algipore did not occur. Particles were degraded during physiologic bone remodeling, and newly formed bone gradually replaced resorbed biomaterial. PMID- 10847963 TI - Long-term results with autogenous onlay grafts in maxillary and mandibular atrophy. AB - This paper describes onlay graft procedures and discusses the technical considerations involved in one- or two-stage implant placement. The results of a 5-year clinical experience with such procedures are reported. Intra- and extraoral donor sites were used to harvest cortico-cancellous grafts: 25 patients were treated with chin monocortical grafts, 8 patients with monocortical hip grafts, and 18 patients with bicortical hip grafts, depending on the size of the defect and the location of the graft. The monocortical grafts were used in the maxilla, and the bicortical grafts in the mandible. Four months after the surgery, a reentry procedure was performed to place implants in the patients with monocortical grafts, while the implants in the mandibular onlays were placed simultaneously with bone augmentation. CT (computer tomography) and Panorax comparisons were made 24 hours postoperatively; after 4 months for the monocortical grafts only; and after 1, 3, and 5 years to evaluate the bone resorption. The results obtained with the onlay grafts suggest that this is the method of choice for solving different kinds of alveolar and basal jaw bone reabsorption. Either a monocortical bone or a bicortical graft (type 1 or type 2 bone atrophy, respectively) with the osseointegrated fixtures-bone graft combination were found to guarantee acceptable reabsorption of a graft during 5 years and prevented failure of the prosthetic rehabilitation. In monocortical grafts, delayed fixture placement by 4-6 months is the most predictable; the one stage procedure is preferred for the bicortical hip graft in the mandible. PMID- 10847964 TI - 2D and 3D computer tomography and sinus graft analysis with various graft materials. AB - Sinus grafting has been a routine surgical procedure since the 1970s. Computer tomography (e.g., SIM/Plant) is available for planning and simulation, but it has not been used to control the success and the long-term results of the planned surgery. The aim of this work was to apply a prospective examination method to determine (a) the extent to which the success of sinus grafting can be followed through 2D (two-dimensional) and 3D (three-dimensional) CT (computer tomography) reconstruction, (b) when choosing this method is worthwhile, and (c) how to visualize incorporating and transforming various graft materials. Twelve patients received 21 sinus graftings, with immediate implantation. Four 2D and 3D CT reconstructions were performed: (1) preoperatively, (2) 1-2 weeks postoperatively, (3) before loading, and (4) 1 year after completing prosthodontic treatment. The sinus grafts were composed of a mixture of HTR Bioplant polymer and autograft, or from a mixture of Algipore (HA, hydroxyapatite) and autograft. In the bilateral cases, both sinus graft mixtures were used, one on each side. The posterior-anterior, the lateral, and the superior-inferior views provided the most useful information; the usefulness of serial images is very limited. Individual 2D and 3D CT reconstructions are proposed, however, in cases of extensive bone atrophy, if complications are expected, if complications arise during surgery, if healing is protracted, or if the documentation is important for other reasons. The long-term results of the two graft materials could be followed up extremely well. Both materials proved outstandingly suitable for sinus grafting. PMID- 10847965 TI - GTR with three different types of membranes in the treatment of intrabony periodontal defects: three-year results in sixty consecutive cases. AB - The aim of the present study was to evaluate the clinical results at 3 years following treating intrabony periodontal defects with different nonresorbable and bioabsorbable membrane barriers. Sixty intrabony periodontal defects were treated according to the principles of guided tissue regeneration (GTR). Twenty pockets were treated with Gore Resolut, a bioabsorbable membrane; 20 were treated with Gore-Tex, a titanium-reinforced membrane; and 20 with nonresorbable Gore-Tex membrane (all manufactured by Gore Regenerative Technologies, Flagstaff, AZ). The therapeutic results were evaluated by assessing probing pocket depth (PPD), recession of the gingival margin (GR), and clinical attachment level (CAL) at baseline, at 1 and at 3 years after therapy. The postoperative phase was uneventful in all cases. At 1 year after surgery, the results showed a mean PPD reduction from 9.42 mm to 3.35 mm (p < 0.0001) with Resolut; from 10.30 mm to 4.00 with titanium-reinforced Gore-Tex (p < 0.0001); and from 8.40 mm to 3.73 mm (p < 0.0001) with Gore-Tex membranes. The mean GR increased from 1.92 mm to 3.70 mm (p < 0.001) with Resolut; from 0.47 mm to 2.85 mm (p < 0.0001) with titanium reinforced Gore-Tex; and from 0.73 mm to 2.15 mm (p < 0.0001) with Gore-Tex membranes. The mean CAL changed from 11.35 mm to 6.92 mm (p < 0.001) with Resolut; from 10.78 mm to 6.85 mm (p < 0.0001) with titanium-reinforced Gore-Tex; and from 9.13 mm to 5.87 mm (p < 0.0001) with Gore-Tex membranes. The clinical results at 3 years were not significantly different when compared with the 1-year results (p > 0.05). No significant differences existed between the mean changes in PPD, GR, and CAL in the three different test groups. Furthermore, one tooth scheduled for extraction for periodontal and prosthodontic reasons was treated with Resolut. Histological analysis 6 months after treatment demonstrated the neoformation of a connective tissue attachment and of new alveolar bone. This additional evidence thus proved that treatment with bioabsorbable membranes according to GTR principles delivers not only clinical improvement, but also histological periodontal regeneration. PMID- 10847966 TI - A new advanced surface modification technique--titanium oxide ceramic surface implants: the background and long-term results. AB - This paper discusses the background of advanced surface modification technologies and presents a new technique that forms a titanium oxide ceramic coating with relatively long-term clinical use. Three general techniques are used to modify surfaces: adding or removing material, or changing material already present. Surface properties can also be changed through laser or electron beam thermal treatment without adding or removing material. The new technique outlined in this paper describes producing a corrosion-resistant, 2000 A to 2500 A thick, coherent crystalline oxide ceramic layer on the surface of titanium implants. The layer is grown electrochemically from the bulk of the metal and modified by heat treatment. Compared with implants covered with other coatings, the improved properties of such oxide ceramic-coated implants include the advantages of higher external hardness and greater adherence between the titanium and the ceramic oxide coating. The virtually perfect insulation between the tissue and the metal prevents metal allergy. The coated implants were subjected to various physical and chemical tests and electron microscopy for a qualitative characterization. Finally, these implants (plates, screws for maxillofacial osteosynthesis, and dental root implants) were followed in surgical practice for 10 years. Tests demonstrated and the acquired experience confirmed the good properties of the titanium oxide ceramic-coated implants. PMID- 10847967 TI - Migration of polyethylene particles around stable implants in an animal model. AB - The aim of this study was to test the hypothesis that a tight seal between bone and implant will eliminate the avenue of particle migration around stable implants. Three types of implants were used in rabbits (polished press-fit Ti-6Al 4V or plasma-sprayed hydroxyapatite [HA]-coated Ti-6Al-4V) or doughy stage polymethyl methacrylate (PMMA). Implants were placed in the condylar notch. Each animal received an intra-articular injection of high density polyethylene (PE) particles (10(8) in 0.4 mL; mean size 4.7 microns) at 4 and 6 weeks postoperatively. Eight weeks postoperatively, peri-implant tissues were examined for PE particles and osteolysis. In all cases, intracellular PE particles were seen at the bone-implant interface and within marrow. No osteolysis was observed. Bone apposition was determined by computerized image analysis. There was no significant difference in the percentage of bone apposition (+/- SD) among the three groups of implants: Ti-6Al-4V (68% +/- 19%), HA-coated Ti-6Al-4V (70% +/- 10%), and PMMA (59% +/- 12%). These results indicate that a polished Ti-6Al-4V surface is as effective as PMMA or HA coating in limiting migration of PE particles around stable osseointegrated implants in rabbits. PMID- 10847968 TI - Solubility changes in shelf-aged ultra-high molecular weight polyethylene acetabular liners. AB - Crosslinking of extruded, air-packaged, irradiated, shelf-aged (10 consecutive years) polyethylene acetabular liners was measured versus time and material location by using a hot xylene extraction protocol. Insolubility (crosslinking) of new polyethylene liners was 87%, but decreased to 45% after 10 years of shelf aging. This degradation is similar to that observed from retrieved (aged in vivo) acetabular cups. Crosslinking varied with depth in the aged liners and with radial location in the unaged liners. Given that crosslinking improves polyethylene's wear resistance, crosslinking degradation of polyethylene orthopaedic components must be controlled to improve the long-term clinical performance of total joint implants. PMID- 10847969 TI - A new method of determining the J-integral fracture toughness of very tough polymers: application to ultra high molecular weight polyethylene. AB - The thrust of the present work was the development of a new modification to the ASTM E813-89 method for determining the J-integral fracture toughness, JIc, of ductile polymers. The advantages and limitations of the modification, vis a vis the ASTM E813-89 and the hysteresis methods, are presented. The modification was then applied to investigate the effect of sterilization method on the JIc of ultra-high-molecular-weight polyethylene (UHMWPE) specimens that were subsequently aged (heated, in a cylindrical pressure vessel, in oxygen gas at 507 kPa and 70 degrees C for 14 d). Relative to the unsterilized material, it was found that gamma-irradiating the polymer leads to a significant drop in its JIc, while sterilization using either ethylene oxide or gas plasma produced no effect on its JIc. The clinical implications of the present results for the in vivo performance of UHMWPE counterfaces, and, hence, hip and knee arthroplasties, are fully discussed. PMID- 10847970 TI - Interfacial strength between molded UHMWPE and PMMA-MMA monomer treated UHMWPE. AB - To solve aseptic loosening of cemented acetabular cup and tibial plateaus of hips and knee joint prostheses in total joint replacement, adhesion between the polymeric prostheses made of ultra-high molecular weight polyethylene (UHMWPE) and bone cement made of polymethyl-methacrylate (PMMA) has been investigated. Previous studies using xylene, PMMA, and MMA monomer solution to treat bulk surfaces of UHMWPE resulted in 11.5 +/- 0.65 MPa as the maximum interfacial strength between the treated UHMWPE and bone cement. The present study eliminates the use of xylene; instead, UHMWPE powders were treated with MMA monomer and PMMA MMA solution, dried, and then compression-molded with untreated UHMWPE powders. The interfacial tensile strengths were measured and the best molding conditions investigated by varying temperature, pressure, and time. Temperature was the predominant factor when compared to that of pressure or time. The maximum interfacial strength between the treated UHMWPE and untreated UHMWPE was 16.3 +/- 0.94 MPa with the molding conditions of 166.5 degrees C, 38.7 MPa, and 1 h. Scanning electron microscopic (SEM) pictures show the coating of the nodules and fibrils of UHMWPE powder with PMMA. Pulled out aggregated nodular space and coated PMMA after fracture could be seen easily from PMMA-MMA treated UHMWPE powder. PMID- 10847971 TI - Pathology of the bone-implant interfaces. AB - Joint replacement surgery has a wide clinical application as a successful technique. However, the release of biomaterial in particulate form from various implant components has been implicated as a cause of two major clinical complications: 1) bone lysis with or without aseptic loosening, and 2) dissemination of wear particles to distant sites with adverse local or systemic cellular responses. This review focuses on the analysis of the clinical material obtained at the time of revision operations and its value in identifying the pathological processes taking place within the bone-implant microenvironment. Important issues discussed include the incidence of infection, characterization of wear particles, particle-cell interactions, local histopathological changes that lead to the formation of erosive inflammatory lesions next to the bone, sensitivity reactions, tumor formation, and the induction of inflammatory factors and cytokines that can influence the rate of bone resorption versus bone formation. PMID- 10847972 TI - Usefulness of dental implants in maxillofacial reconstruction. AB - Fabricating maxillofacial prosthesis can be challenging. Placement of implants can have a dramatic effect on the stability and retention of the prosthesis in patients. This article provides clinical retrospective analysis of osseointegrated implants used for maxillofacial reconstruction. Patient charts and radiographs were reviewed to determine implant status, stability of prosthesis, and masticatory and speech function. Of the 104 implants placed, there were 4 implant failures. The overall survival rate for implants in this patient population was 96.1%. The stability of maxillofacial prostheses demonstrated significant improvement after anchorage to implants. Implant-borne maxillofacial prostheses are better stabilized and retained than non-implant borne prostheses, providing an improved quality of life to patients requiring prosthesis rehabilitation of maxillofacial defects. PMID- 10847973 TI - Innovative techniques in arthroscopic meniscal repair. AB - Advanced techniques for arthroscopic repair of meniscal tears offer new ways to repair the meniscus and reduce the rate of long-term arthrosis. The new all inside technique reduces complications, morbidity, and surgical time as compared to the inside-out or outside-in methods of repair. Application of biomaterial technology to new implants used for all-inside arthroscopy show early promising clinical results. The purpose of this report is to describe new biomaterials and techniques being used successfully in arthroscopic surgery on the meniscus. PMID- 10847974 TI - Modern concepts in arthroscopic Bankart repair. AB - The purpose of this report is to review recent advances in Bankart repair that allow it to be performed arthroscopically. Metallic implants in the form of staples, screws, or rivets were used initially, but problems with loosening of implants has prompted development of alternative devices and techniques. A modified form of the transglenoid suturing technique used in open Bankart repair has been used successfully but is associated with an increased risk of soft tissue complications. The use of absorbable tacks avoids some of the complications associated with this approach, but the ability to advance the capsule to reduce the volume is limited. Recently, the use of suture anchors has been described for arthroscopic Bankart lesion repair. However, the ability of these anchoring systems to advance and reduce the capsular volume is limited, and the arthroscopic techniques are technically demanding. One particularly innovative suture anchor system uses a bioabsorbable suture anchor with an integrated suture passing system. This self-contained implant allows multiple passes with suture through the capsule, followed by attachment to the glenoid with the bioabsorbable anchor. This procedure allows the capsule to be advanced by passing suture through before it is anchored to the glenoid, thereby reducing capsular volume to help restore shoulder stability. PMID- 10847975 TI - Rotatory motion of a cemented acetabular component caused by aseptic loosening and dissociation of the polyethylene liner: a case report. AB - A 63-year-old woman without rheumatoid arthritis who had undergone cemented total hip arthroplasty developed coxalgia 1 year later. Radiography showed a clear zone at the cement-implant interface and rotatory motion of the acetabular component, so revision surgery was performed 4.5 years after the first operation. Aseptic loosening was noted and 7 spacers outside the acetabular component were broken and buried under the cement mantle. We consider that the rotatory motion may be caused by circumferential loosening of the acetabular component stemming from incomplete implant fixation and also dissociation of the ultra-high-molecular weight polyethylene (UHMWPE) liner from the metal base plate. PMID- 10847976 TI - Macroporous calcium phosphate ceramic: a prospective study of 106 cases in lumbar spinal fusion. AB - Macroporous biphasic calcium phosphate (MBCP, Triosite) is well known for its safety, absence of allergenicity, and excellent bone-bonding capacity, and it has been widely used as a bone graft substitute in orthopaedic, ENT, and dental surgery. This study investigates the clinical performance of this synthetic porous ceramic in a series of 106 patients, mainly with degenerative spine aetiologies (95/106) and with a minimum follow-up of 2 years. All patients were treated with posterior correction involving the semi-rigid New Orleans instrumentation. Spinal fusion was always performed using MBCP granules mixed with autogenous bone chips and bone marrow obtained from the local spine. Fusion of the spine was confirmed for 100 patients, and 6 non-unions were observed (3 resulting from primary spondylolisthesis). This study shows that MBCP provides suitable results in spinal fusion involving a semi-rigid instrumentation. Because the indication of degenerative spine is not very favorable to fusion, this technique appears to be a good alternative to autografts and could decrease patient morbidity resulting from iliac bone grafting. PMID- 10847977 TI - Distraction osteogenesis of the midface: a new implantable distraction device. AB - This report describes the use of a new implantable device for distraction osteogenesis of the maxilla in treatment of midface hypoplasia seen in craniofacial syndromes. The technique of distraction osteogenesis is a recent advance in plastic surgery for treatment of bone growth disorders. The device is totally implantable except for the activating pin and can provide up to 30 mm of total advancement. We will describe our surgical technique and distraction protocol, illustrated by a case report of the device successfully used in a child with Crouzon syndrome. PMID- 10847978 TI - Fostering empowering participation in prevention programs for disadvantaged children and families: lessons from ten demonstration sites. AB - Empowering participation is relevant to many prevention programs. Yet many attempts at empowerment proceed without any clear understanding of the nature of empowerment or the practical requirements of an empowering process. This paper draws on the extensive program-development research for two Canadian prevention projects for disadvantaged children and families: the Parent Mutual Aid Organizations in Child Welfare Project and the Better Beginnings. Better Futures Prevention Project. The lessons learned in these projects about fostering empowering participation are summarized. The paper emphasizes the necessity to respect the demanding and complex requirements of successful empowering participation as well as the need to balance empowering participation with other priorities in prevention programs and projects. PMID- 10847979 TI - [Psychosocial adjustment to asthma and juvenile diabetes mellitus in school-age children]. AB - The main purpose of this study was to compare psychological and social problems among chronically ill school-aged children to those observed in a group of healthy children of the same age. On the basis of the data collected, it was also possible to compare self-perception and social functioning problems between the 2 groups of children, as well as to identify the contributing factors explaining the observed differences in psychosocial adaptation between these 2 groups. Three measures were used to assess psychological and social problems. Mental health problems were measured by the Dominique (Valla, Bergeron, Berube, Gaudet, & Saint Georges, 1994). Self-Perception and Social Functioning were assessed by the Self Perception Profile (Harter, 1985) and the Rating Scale of Child's Actual Behavior (Harter, 1985). Information on characteristics of the child and his or her family was also collected. Sixty children aged 6 to 11 and diagnosed as having juvenile diabetes or asthma, 59 control children, and the children's parents participated in this study. A chi-square test and multivariate analysis of variance were used to compare the 2 groups of children. Results indicate that children with diabetes and asthma do not present more psychological and social problems than other children of the same age. Results also suggest that age is the only variable indicating a higher proportion of mental health problems among chronically ill children. Results are discussed and compared to those of previous studies from which the conceptual frame-work of this study was developed. PMID- 10847980 TI - From housing to homes: a review of the literature on housing approaches for psychiatric consumer/survivors. AB - In this paper, we review the literature on housing for psychiatric consumer/survivors since the publication of the Nelson and Smith Fowler (1987) review more than a decade ago. First, we review research and propose a definition to contrast key features of three approaches to housing: (a) custodial, (b) supportive, and (c) supported. Second, we examine studies of the relationships between the characteristics of housing and adaptational outcomes for residents (e.g., personal empowerment). Third, we review studies which have examined out comes for residents for these three different housing approaches. We conclude by critically reflecting on the values and research of the different approaches to housing, to make recommendations for future policy and planning, practice, and research. PMID- 10847981 TI - An evaluation of a community-based vocational rehabilitation program for adults with psychiatric disabilities. AB - The purpose of this research is to examine the effectiveness of a project aimed at the vocational rehabilitation of individuals suffering from chronic psychiatric disabilities. Gastown Vocational Services (GVS) is a specialized vocational rehabilitation program, under the auspices of Greater Vancouver Mental Health Service Society. The project consisted of three distinct phases and utilized a gradual, step-by-step rehabilitative approach to achieve vocational success. The first phase of the GVS project included comprehensive vocational assessment and work-readiness skill training. Participants in this phase met in small groups for three hours, three times a week for a 12-week period. The second phase involved supported work-experience placements in the community. These placements were two to five months in duration. The final phase included assistance in seeking employment, job re-training, or educational programs. Assessment measures were taken before participants began the program, immediately after the 12-week job preparation program, and at six-month follow-up. Seventy three individuals participated in the training program over a two-year period. Their progress was compared to 18 individuals comprising a Waiting List Control group. The results showed significant improvement in the Intervention group on measures of assertiveness, work behaviour, depression, income, and employment status. No changes were evident in the Waiting List Control group. PMID- 10847982 TI - [Intervention with adjudicated drug-dependent patients: professionals and judicial professionals]. AB - Many studies indicate that a large number of addicts commit crimes and, conversely, many offenders are addicts. Individuals who fall into both these categories show clear bio-psychosocial problems. With few resources at their disposal, professionals from both the Department of Health and Social Services and the Department of Public Security feel uncomfortable trying to help these individuals. This study presents the results of 6 focus groups on the types of services that should be offered to addict-offenders. Results indicate that proper assessment of the addiction and the addict-offender's motivation to undertake rehabilitation is essential. Another crucial element is an open channel of communication among the various professionals involved in the case management and treatment of these individuals. Finally, a good reference system is needed. However, the authors insist on the caution that must prevail when control and help are provided together in a concerted effort. PMID- 10847983 TI - Standards for batterer intervention programs in Canada: a history and review. AB - This article reports on the growing trend of setting standards for intervention with batterers. All existing Canadian standards for batterer intervention programs (BIPs) are examined, including a discussion on how they came into effect and any controversy associated with their development. A brief background of the battering intervention field and the growth of BIP standards in North America is provided, followed by a description of the types and content of Canadian standards. Compliance with standards were mandatory in two regions and voluntary in three others. Two more regions had standards in a draft stage of development. Elements of standards typically included directives on: general program goals; the protocol and procedures programs should follow, including counselling format; and program staff ethics and qualifications. The article concludes with a discussion of the findings and suggestions for improving standards. PMID- 10847984 TI - Three perspectives on the rehabilitation needs of consumers. AB - Empowering individuals with serious mental health problems includes providing services which they themselves identify as needs. This paper reports on a study which examined needs for rehabilitation services from the perspectives of consumers with chronic mental health problems, their significant others, and case managers. In most instances far greater needs for services were expressed by consumers and significant others than by case managers, indicating that case managers pay insufficient attention to what consumers want. Professional control and power imbalance between case managers and consumers likely contribute to this situation. Study findings have been used to initiate change at the organizational level and at the level of individual professionals. PMID- 10847985 TI - [Contribution of individual characteristics to learning by children with first exposure to a prevention of abuse program]. AB - This study explores the contribution of personal characteristics in the knowledge and skills gains of children taking part in the child abuse prevention program ESPACE. The program is an adaptation of the widely implemented American Child Assault Prevention Program (CAP). A group of 107 fourth-grade children completed a knowledge questionnaire and a video vignette measure designed to evaluate preventive skills in abusive and potentially abusive situations. Children were also invited to complete questionnaires evaluating individual characteristics such as self-esteem, locus of control and self-efficacy. The results highlight the importance of certain individual variables such as the child's gender, self efficacy and perception of athletic competence in the acquisition of sexual abuse preventive skills. PMID- 10847986 TI - Tough choices: women, abusive partners, and the ecology of decision-making. AB - In a qualitative study, 35 women who had lived with violent partners were interviewed regarding their experience making decisions about their relationships to their abusers. This paper explores their stories, integrating the common themes with reports from other studies. The insights of the women participating in this and other research support an ecological understanding of the tough choices they face--a framework which respects the role of environmental factors (demands and resources) in helping or constraining choices at the same time as it recognizes the importance of powerful mediating factors (beliefs, values, and a sense of personal efficacy). PMID- 10847987 TI - Children's needs in shelters: mother's perceptions. AB - The needs of children who witness violence directed at their mothers have only recently gained attention. While shelter services to address children's responses are increasingly evident, there is still much we do not understand about their plight, particularly from the perspective of their mothers. This study used focus group methodology to identify mothers' perceptions of their children's needs while they were in shelters. Five focus group sessions were held with 27 mothers who had experienced violence in their homes and had used shelter services within the previous two-year period. Data were analyzed to identify six dominant categories of need and 12 supporting thematic categories. The dominant categories included: the counselling needs of the children; day-to-day living needs, such as maintaining family integrity and normality; recreational needs; the needs of children with respect to parenting; and finally, aspects of shelter support for mothers, and the impact of the shelter environment on children. The implications of the findings for program development are discussed. PMID- 10847988 TI - [Evaluation of a program designed to develop social competence in kindergarten children]. AB - This paper presents an evaluation of the implementation and impact of a program promoting social competence among kindergarten children. This program was conducted experimentally in 8 schools (10 classrooms, n = 165) in Montreal; 5 classrooms (n = 74) from 5 other schools formed the control group. Analysis of the impact reveals significant gains in self-esteem and conflict resolution skills in the experimental group compared to the control group. However, there was no significant change in pro-sociality and social withdrawal. Finally, an unexpected result was found in the area of boys' aggression. Analysis of the implementation indicates that teachers found the program's activities easy to use and adapt. In addition, great diversity in the utilization of these activities was observed. These results lead to areas of discussion concerning: (a) the links between gains realized by children and the emphasis teachers place on some aspects of the program; (b) the development of social competence among preschoolers; (c) the evaluation of the this type of program. PMID- 10847989 TI - DMEC. Disability Management Employer Coalition. PMID- 10847990 TI - Therapists have future wage trends in their own hands. PMID- 10847991 TI - Components of a cardiac rehab program. PMID- 10847992 TI - Preventing and treating bone loss. PMID- 10847993 TI - Preventing fraud and abuse. PMID- 10847994 TI - The managed care competitor. PMID- 10847995 TI - Settling down to business. PMID- 10847996 TI - Relief arrives for rehab providers. PMID- 10847997 TI - Polio survivors return to battle. PMID- 10847998 TI - What gets measured gets done. PMID- 10847999 TI - The healing properties of water. PMID- 10848000 TI - A Polish approach to CP. PMID- 10848001 TI - Riding out the storm ahead. PMID- 10848002 TI - Raising the standards. PMID- 10848003 TI - Congressional testimony highlights: potential and barriers. PMID- 10848004 TI - New videoconferencing technologies and applications. PMID- 10848005 TI - Health records on the Internet: a harbinger of massive changes in healthcare. PMID- 10848006 TI - Sleepless in Edinburgh. Getting new products certified in the U.S. and in Europe. PMID- 10848007 TI - Review of standalone telemedicine software. PMID- 10848009 TI - Telemedicine in federal prisons. PMID- 10848008 TI - Clinical data repository: one stop shopping for data. PMID- 10848010 TI - House calls through the Web.... PMID- 10848011 TI - Two prison telemedicine programs: California and Texas. PMID- 10848012 TI - Videoconferencing 2000. PMID- 10848013 TI - Telemedicine helps with Hurricane Floyd disaster relief. PMID- 10848014 TI - Online health information triggers legal repercussions. PMID- 10848015 TI - Embracing mobile personal video. PMID- 10848016 TI - Smart card telemedicine. PMID- 10848017 TI - Application of telemedicine in home health services in Korea. PMID- 10848018 TI - Remote access: the last mile is getting shorter. PMID- 10848019 TI - China telemedicine review. PMID- 10848020 TI - A cash cow for rural telemedicine. PMID- 10848021 TI - Annual survey. Teleradiology service providers. PMID- 10848022 TI - Office of inspector general issues second telemedicine opinion. PMID- 10848023 TI - The embedded future of telemedicine. PMID- 10848025 TI - Simplification: standards for privacy of health information. The implications for telemedicine systems. PMID- 10848024 TI - And now ... e-prevention. PMID- 10848026 TI - Kaiser Permanente Tele-Home Health Research Project. AB - In summary, the quality of care and satisfaction measures of the two systems are remarkable for what they are not--that is, not different. They suggest no difference between a culturally accepted model of care, with roots extending deep into human experience, both personal and traditional, and one based on a radical departure therefrom--care in the physical absence of a carer. They suggest, they do not indicate, because the study was not designed as a direct comparison. Numbers were small. Other flaws are evident. But Barb Johnston and her colleagues have opened the door for larger groups with deeper pockets to prove to conclusion what they have suggested--that telecare works. They have succeeded in providing telecare with what it has hitherto lacked: a solid suggestion of equivalence with the established system, and perhaps for markedly less cost. And they have done so in the face of difficulties and barriers which may have prevented them from achieving even better results. The study is a notable achievement, and its authors and Kaiser Permanente deserve the gratitude of the entire telemedicine community. PMID- 10848027 TI - Telehealth and traumatic brain injury: creative community-based care. PMID- 10848028 TI - Hope for the 21st century: health is a multilateral interest. PMID- 10848029 TI - Predicting the potential for risky behavior among those "too young" to drink as the result of appealing advertising. AB - A survey of 273 children in Washington state used a predrinking behavior index as a behavioral outcome to assess media effects on precursors to drinking among children for whom alcohol consumption is not yet occurring. It also examined age trends in relevant beliefs and behaviors. Perceptions of advertising desirability, the extent to which it seemed appealing, increased steadily from third to ninth grade, whereas identification with portrayals, the degree to which individuals wanted to emulate portrayals, leveled off after sixth grade. Expectancies, positive social benefits perceived to be associated with drinking alcohol, also increased with age, particularly between sixth and ninth grade. When demographics and grade level were controlled, desirability predicted identification, and both predicted expectancies, which is consistent with media decision-making theory. Expectancies correlated with alcohol predrinking behavior, and expectancies predicted risky behavior, with demographics and grade level controlled. Predrinking behavior and reported risky behavior were correlated. The results provide cross-sectional support for the view that beliefs and desires developing by third grade prime children for future decisions regarding substance use. PMID- 10848030 TI - Does knowledge make a difference? The association between knowledge about genes and attitudes toward gene tests. AB - A survey was conducted (n = 1,216) to evaluate the level of knowledge about basic facts of genetics and attitudes toward gene tests among the Finnish population. Our results show that a majority of the Finnish population generally approved of gene tests. Well over half of the respondents believed that genetic testing will have positive consequences, but many also had worries related to the development of gene tests. We showed that there was an association between knowledge and attitudes, but better knowledge did not simply lead to unambiguous acceptance. Among those with the highest level of knowledge there was in many respects both more enthusiasm and more skepticism than among those with the lowest level of knowledge. Those with a low level of knowledge had more difficulties in taking a stance toward attitude statements. In general, steps should be taken, first, to increase citizens' capabilities to seek and understand information about genetics and specific diseases when faced with a need for it. Second, steps should be taken to provide citizens with tools to perceive and discuss potential ethical and social problems, which development of gene technology, genetic testing, and screening bring along. Sites for these enterprises could be the biology school curriculum, genetic counseling, media, as well as health education and health communication in general. PMID- 10848031 TI - Social cognitive factors associated with mother-adolescent communication about sex. AB - To better understand why some mothers talk to their children about sex and others do not, we examined the role of two social cognitive variables--self-efficacy and outcome expectancies--in explaining sex-based communication. The present study was part of a larger study to test the efficacy of two HIV prevention programs for mothers and their adolescents. Mothers and their adolescents were recruited from a large community organization that serves youth who live in disadvantaged circumstances. The sample for the present study included 486 mothers who averaged 38.4 years of age (SD = 6.73). The majority were African American (97.7%), not married (66.7%), and had a high school degree (89.5%). Their adolescents ranged in age from 11 through 14 years of age and most were male (61.3%). The results of the analysis revealed that mothers who expressed higher levels of self-efficacy and more favorable outcomes associated with talking to their children about sex were more likely to do so. In a regression analysis, we learned that the mother's degree of efficacy beliefs, along with her expected outcomes associated with talking about sex, the importance of religious beliefs to her, and the age and sex of her adolescents were important factors associated with talking with them about sex. PMID- 10848032 TI - Weaving meanings together in an AIDS residence: communicative practices, perceived health outcomes, and the symbolic construction of community. AB - This study examines the role of communicative practices in the symbolic construction of health and community among residents living in a residential facility for people with AIDS. Questionnaire results show that two types of communicative practices--governance/support and everyday/special--are significantly related to residents' perceived physical and emotional health outcomes, their perceptions of the residence as a community (as opposed to a facility or institution), and their satisfaction with living there. Everyday/special practices reveal the communicative means by which perceived health is socially constructed, while governance/support practices reveal the importance of meta-communication for massaging the issues and tensions that pervade community life, and both sets of communicative practices help explain members' perceptions of the residence as a community and their satisfaction with living there. The study provides additional evidence regarding the constitutive nature of communication as a connecting thread that weaves together individual and collective meanings to help create and sustain such concepts as health and community. PMID- 10848033 TI - Response to DeJong and Wallack's article: a critical perspective on the drug Czar's anti-drug media campaign. PMID- 10848034 TI - The drug Czar's anti-drug media campaign: continuing concerns. AB - In reviewing the anti-drug media campaign organized by the Office of National Drug Control Policy (ONDCP), we expressed concern about both its strategic direction and execution (Journal of Health Communication, 1999, 4:2 155-160). The response from ONDCP's director, General Barry R. McCaffrey, still leaves us concerned about the campaign's chances for success. We review ONDCP's strong opposition to including alcohol in the campaign, its continued reliance on fear appeals, and its relative inattention to building support for environmental prevention strategies. We voice again our concerns about ONDCP's underutilization of an expert review committee of behavioral scientists. PMID- 10848035 TI - Adolescents' perceptions of print cigarette advertising: a case for counteradvertising. AB - Cigarette smoking is a major preventable cause of serious chronic disease. The majority of smokers begin smoking during their adolescent years. Print cigarette advertising plays an influential role in encouraging young people to smoke. Eight hundred and forty-three middle/junior high-school-aged students from a large midwestern metropolitan area participated in this study of adolescents' recognition of and belief in messages associated with print cigarette advertising. Results indicated no significant difference between smokers and nonsmokers in their recognition of messages associated with print cigarette advertising. However, smokers and nonsmokers differed significantly in the degree to which they believed the messages communicated by print cigarette advertising. Consistent with social cognitive theory, students who smoke at least occasionally were more likely to believe messages conveyed by print cigarette ads than were students who had never smoked. Implications of these findings for research and practice are discussed, including specification of guidelines for effective counteradvertising. PMID- 10848036 TI - How physician organizations are responding to managed care. AB - Despite a rash of troubles in 1998, physician organizations--formed in response to managed care plans--can thrive if they are locally owned, physician-run and rationally sized, said panelists at a recent roundtable organized by the Center for Studying Health System Change (HSC). The panelists also noted the market's slow progress toward global capitation as a way of compensating physician organizations, and the generally weak state of information systems required to support the goals behind capitation: accountability, efficiency and quality. This Issue Brief reports on governance, physician-hospital relationships, capital needs, compensation and other developments covered at the roundtable. PMID- 10848037 TI - Bilateral prophylactic mastectomy in women with an increased risk of breast cancer. PMID- 10848038 TI - Hyperbaric oxygen therapy for wound healing. PMID- 10848039 TI - Salivary estriol for the assessment of spontaneous preterm labor. PMID- 10848040 TI - Salvage high-dose chemotherapy with allogeneic stem-cell support for relapse following high dose chemotherapy with autologous stem-cell support for solid tumors. PMID- 10848041 TI - Serum antibodies for the diagnosis of inflammatory bowel disease: ANCA for ulcerative colitis and ASCA for Crohn's disease. PMID- 10848042 TI - Small bowel transplants in adults and multivisceral transplants in adults and children. PMID- 10848043 TI - Thalidomide: new preparation. For well-defined indications. AB - (1) Thalidomide is highly teratogenic. It can also cause irreversible neuropathy. (2) In type II lepra reactions evidence of efficacy at a dose of 400 mg/day is based on a few relatively old comparative trials and above all a review of more than 4,500 cases. Most patients feel a benefit within 24-48 hours after beginning the treatment, but most also require lengthy treatment at a reduced dose (25-100 mg/day) to avoid relapses. (3) Two placebo-controlled trials involving a total of 130 patients have shown the efficacy of thalidomide in the treatment of severe recurrent aphthous disease. (4) In Jessner-Kanoff disease, a mild condition, the efficacy of thalidomide has been shown in a single placebo controlled trial. (5) In chronic lupus erythematosus, non comparative trials have shown remission from skin lesions in patients resistant to previous treatments. (6) In the curative treatment of chronic graft-versus-host disease in bone marrow graft recipients, thalidomide has been assessed only in non comparative trials involving a few dozens of patients. PMID- 10848044 TI - Sildenafil and erectile dysfunction: new preparation. Helpful. AB - (1) The clinical file is bulky, including 14 double-blind, placebo-controlled trials conducted in normal conditions of use. (2) Sildenafil effectively treats the symptoms of erection disorders, whatever their origin (psychogenic or organic, especially spinal injuries). Efficacy seems slightly lower in case of diabetes or total prostatectomy. (3) Overall, sildenafil allows 80 to 90% of patients to have an erection adequate for sexual intercourse, but only one in two sexual acts on average were considered "satisfactory" by the clinical trial investigators. (4) Sildenafil does not affect sexual desire. It has not been studied in men without erection disorders. The results of ongoing trials in women are not yet known. (5) The adverse effects of sildenafil seem infrequent and generally mild. However, the risk of sudden arterial hypotension if the drug is combined with nitrate derivatives calls for careful safety monitoring. This combination is contraindicated. (6) In our opinion sildenafil should not be prescribed to anyone with cardiovascular risk factors or a history of heart problems. PMID- 10848045 TI - Latanoprost: new preparation. Antiglaucoma eye drops that can change the colour of the iris. AB - (1) Latanoprost, a prostaglandin F2 alpha analogue, is an antiglaucoma drug. (2) The clinical file is fairly thorough. (3) Dose-finding studies show that the optimal daily dose is a single drop of 0.005% solution, preferably in the evening. Two drops a day are less effective than one drop a day. (4) The local hypotensive action of latanoprost persists in the long term (current follow-up one year). (5) Four double-blind trials have compared 0.005% latanoprost eye drops and 0.5% timolol eye drops. In three trials the effect of latanoprost on intraocular pressure was statistically stronger than that of timolol. However, the difference in mean intraocular pressure was less than 2 mm Hg between the two treatments, and the clinical relevance of such a difference is not known. (6) Latanoprost has not been compared with the other available antiglaucoma eye drops. (7) Various trials have shown that intraocular pressure is statistically lower when latanoprost is combined with another antiglaucoma eye drop preparation (timolol, pilocarpine or dipivefrine) than during monotherapy. The additive action of latanoprost eye drops when combined with oral acetazolamide has been established more soundly in a comparative double-blind trial. (8) In approximately 30% of cases (especially patients with non homogeneous eye colour), latanoprost eye drops can cause permanent darkening of the iris. Patients must be warned of this risk before beginning treatment. PMID- 10848046 TI - Biogenetic glucagon: new preparation. Four times more expensive for the same service. AB - (1) Human biogenetic glucagon has replaced traditional glucagon of animal origin. It is the same substance but is obtained through a different manufacturing process. (2) The file on the efficacy and safety of biogenetic glucagon is limited. It suggests nevertheless that the hyperglycaemic effect of biogenetic glucagon is equivalent to that of animal glucagon and that the underlying biotechnology does not introduce new risks. (3) The main indication for glucagon is in the treatment of severe hypoglycaemia in insulin-treated diabetic patients. In this indication glucagon is usually administered by persons with no medical training (relatives) who must be taught when and how to use the drug appropriately. (4) Glucagon has been used for more than 25 years in the treatment of acute betablocker intoxication. Despite the absence of randomised trials most authors consider it superior to beta-adrenergic agonists in severe cases. PMID- 10848047 TI - Somatropin in adults: new preparation. No benefit. AB - (1) Growth hormone deficiency in adults is a poorly defined disease, and its impact in terms of morbidity and mortality is not known. (2) The assessment files on the three somatropin ranges are based on results for nearly 600 adult patients included in placebo-controlled trials. (3) While clinical trials are relatively numerous, their quality is mediocre. Only methodological trickery enabled the authors to elicit significant results using surrogate end points; no benefit of somatropin on tangible clinical criteria has been proven. (4) At the beginning of treatment somatropin frequently has adverse effects linked to water and sodium retention; the risk of severe adverse effects has not been ruled out (diabetogenic or carcinogenic effects). (5) Treatment necessitates chronic daily injections. PMID- 10848048 TI - Benazepril and the kidney: new indication. No better than the other ACE inhibitors. AB - (1) The action of benazepril on the progression of renal failure in non diabetic patients has been assessed in a comparative placebo-controlled trial involving nearly 600 patients, most of whom were hypertensive. (2) This trial showed a statistically significant slowing of renal failure on benazepril, but the possibility of bias casts doubts on this conclusion; moreover, excess deaths occurred in the benazepril group. (3) Currently, the slowing of the progression of renal failure is not a valid basis for choosing between angiotensin-converting enzyme inhibitors for non diabetic hypertensive patients with renal failure. PMID- 10848049 TI - Drug interactions: 1998 update. AB - (1) The 1998 edition of the Drug Interactions section of the French data sheet compendium (Dictionnaire Vidal) includes welcome updates. (2) Increasingly numerous interactions involve tramadol, clarithromycin, oral anticoagulants, antiepileptics, theophylline and various psychotropic agents. (3) An entire section is now devoted to the numerous potassium-raising drugs. PMID- 10848050 TI - Isotretinoin and skin lesions during wax epilation. AB - (1) Several cases of sometimes extensive skin lesions caused by epidermal stripping during wax epilation have been observed in women on oral isotretinoin. (2) Patients taking isotretinoin must be warned that their skin may be fragile and that they should thus avoid wax epilation. PMID- 10848051 TI - Bromide intoxication. AB - (1) A possible cause of asymptomatic "hyperchloraemia" or hyperchloraemia associated with neurological disorders. (2) Bromide-based preparations must not be prescribed or dispensed. PMID- 10848052 TI - Loss of the epileptic aura on lamotrigine? AB - Some epileptic patients, who used to experience auras, have been injured during unexpected seizures while on lamotrigine. PMID- 10848053 TI - Melatonin: interesting, but not miraculous. AB - (1) In the United States melatonin is just a dietary supplement, but in Europe its status varies from country to country and also over time. It is illegal in some European member states but tolerated or authorised as a drug or dietary product elsewhere. Melatonin, a hormone secreted by the pineal gland, has been on the front cover of magazines throughout the world for its claimed effects on ageing, cancer and many other health problems, opening up a vast potential market. (2) Only its use in jet lag, sleep disorders and advanced cancer has been tested clinically (albeit scantily). (3) Melatonin seems to alleviate jet lag symptoms, but that could be linked to its moderate hypnotic effect. (4) The use of melatonin to treat major insomnia cannot be envisaged until its long-term safety has been proven. With this proviso, and if efficacy is confirmed in sufficiently large comparative trials, melatonin could prove useful for treating major sleep disorders in some patients, especially blind people and those with severe neurological disabilities. (5) According to open trials conducted by a single team, melatonin, alone or combined with interleukin-2, could slightly lengthen the survival of patients with some advanced cancers, but even partial tumour remissions are rare. (6) All other "indications" are based on simplistic hypotheses or purely commercial considerations. PMID- 10848054 TI - 19th French pharmacovigilance meeting. Health professionals and consumers are inadequately informed. AB - (1) Drug excipients can have adverse effects independently of the active substance. This should be borne in mind when a patient suffers recurrent adverse effects with drugs of different classes. (2) Elderly patients are particularly at risk of adverse effects when they take a large number of drugs or when the prescriber fails to estimate creatinine clearance. (3) At the end of pregnancy drugs taken by the mother can have adverse effects on the fetus and neonate, such as renal impairment with nonsteroidal antiinflammatory drugs, and a withdrawal syndrome with high-dose buprenorphine. (4) The incidence of Reye's syndrome in France is now comparable to that in countries where paediatric use of aspirin is highly restricted. (5) The lack of information on adverse drug effects aimed at the public and health professionals is worrying. Undernotification of adverse effects by health professionals needs to be addressed. PMID- 10848055 TI - The Erice declaration: on communicating drug safety information. PMID- 10848056 TI - Carvedilol: new preparation. Encouraging but inadequate data. AB - (1) Carvedilol is the first betablocker to obtain approval in France for the treatment of heart failure. (2) The improvement in haemodynamic parameters was initially documented in three methodologically sound studies involving a total of 156 patients, with follow-up limited to 16 weeks. (3) A placebo-controlled, double-blind trial involving 1,094 patients showed a beneficial impact on overall mortality of 4.6% in absolute terms after a median follow-up of 6.5 months. This benefit was not found in another trial involving 415 patients followed for 19 months on average. Results for symptom-based criteria are conflicting. (4) When treatment is introduced very gradually, adverse effects seem to be minor (malaise) and infrequent. (5) The place of carvedilol in the treatment of heart failure is not yet precisely documented. PMID- 10848057 TI - Zidovudine + lamivudine: new preparation. Fewer tablets required. AB - (1) Combivir is a fixed-dose combination of two HIV reverse transcriptase inhibitors: zidovudine (300 mg) + lamivudine (150 mg). (2) A meta-analysis of four trials shows that this combination is more effective clinically than zidovudine monotherapy. One trial also shows that clinical efficacy is even better with the triple combination of the protease inhibitor indinavir with zidovudine + lamivudine. (3) The risk of anaemia and neutropaenia necessitates blood cell monitoring during treatment. (4) Compared with lamivudine and zidovudine taken separately, Combivir does not reduce the frequency of doses (two a day), but the daily number of tablets falls from four to two, provided that the patient does not require dose adjustment. PMID- 10848058 TI - Zolmitriptan: new product. Similar to sumatriptan. AB - (1) Zolmitriptan is an antimigraine drug similar to sumatriptan. (2) The clinical file mainly comprises placebo-controlled dose-finding studies recommending an optimal oral dose of 2.5 mg. (3) Zolmitriptan has been compared with sumatriptan in a trial that showed no difference in efficacy. In particular, the recurrence rate of headache after initial relief was not lower on zolmitriptan than on sumatriptan. (4) The safety profile of zolmitriptan is similar to that of sumatriptan. The contraindications relating to a history of cardiovascular disease must be respected because of the vasoconstrictive effect of the drug. (5) Zolmitriptan has the same drug interactions as sumatriptan. Moreover, zolmitriptan should not be used during migraine attacks by patients on propranolol. PMID- 10848059 TI - Clofazimine: new products. Major antileprous drug. AB - (1) Clinical trials have shown that clofazimine monotherapy is effective. To avoid the emergence of resistance, however, the WHO reference treatment since 1982 has been the rifampicin + dapsone + clofazimine combination. (2) Epidemiological studies show that this polychemotherapy regimen is effective, but we do not yet know if treatment should be continued until skin smears become negative, or whether a 2-year course suffices. (3) Adding clofazimine to the polychemotherapy regimen partly prevents and also lessens the intensity of lepra reactions. However, in the absence of comparative trials, we do not know the part played by clofazimine in the efficacy of the WHO-recommended three-drug regimen. (4) The main adverse effects of clofazimine arise from its accumulation in the skin and eyes, where it frequently causes hyperpigmentation that resolves very slowly on treatment cessation. PMID- 10848060 TI - Rivastigmine: new product. No better than donepezil. AB - (1) Rivastigmine is the third anticholinesterase to be approved for treating Alzheimer's disease, after tacrine and donepezil. (2) The clinical evaluation dossier comprises only 3 double-blind, placebo-controlled trials of good methodological quality. (3) Rivastigmine showed some effect on the scores of validated scales, but the clinical relevance of this is doubtful. Only 4% of patients are likely to benefit from rivastigmine 6-12 mg daily. Clinical outcome after 6 months of treatment is not known. Note that these trials were not published in detail. (4) Cholinergic reactions have been reported. Note that for donepezil, no adverse effects on the liver have been observed. PMID- 10848061 TI - Iloprost: new indication. Not adequately assessed. AB - (1) Iloprost, a vasodilatory prostacyclin analogue administered by infusion, is indicated for second-line therapy in patients with severe ischaemia of the lower limbs, when surgical revascularisation fails or is contraindicated. (2) In thromboangiitis obliterans the clinical file on iloprost has remained inadequate since the product was first released. (3) A meta-analysis of 6 clinical trials giving conflicting results in patients with stage III or IV lower-limb arterial disease favoured iloprost. But the results of this meta-analysis are uninterpretable because of methodological biases. It is not known what effects iloprost has in the short term (on pain and skin damage) or in the long term (on the risk of amputation). (4) The iloprost dose must be adjusted individually according to adverse effects linked to vasodilation. PMID- 10848062 TI - Welcome withdrawal of amineptine in France. PMID- 10848063 TI - Hepatitis B vaccine and demyelination (cont'd). AB - (1) New data on adverse events potentially linked to vaccination against hepatitis B in France were made public in October 1998. (2) The incidence of spontaneous notifications of new cases of central demyelination during the period 1990-1997 was about 1 case per 100,000 hepatitis B vaccinees. (3) Preliminary results of two case-control studies show no statistically significant increase in the risk of central demyelination after hepatitis B vaccination. (4) A risk of neurological complications linked to vaccination against hepatitis B cannot be ruled out, but, at worst, must be very small. PMID- 10848064 TI - Fat distribution and HIV protease inhibitors. AB - (1) Disturbing changes in body fat distribution started to be described in 1997 in HIV-infected patients treated with antiretroviral drugs, with fat accumulation at various levels of the trunk, and thinning of the limbs, buttocks and face. (2) The vast majority of cases occurred in patients on HIV protease inhibitors. (3) The long-term clinical consequences are unknown. PMID- 10848065 TI - Tolcapone withdrawn. PMID- 10848066 TI - Male infertility due to anabolic steroids. AB - (1) The anabolic steroids used by some sportsmen can cause azoospermia, which usually resolves when the drugs are stopped. (2) Infertile sportsmen should be asked if they use anabolic steroids. PMID- 10848067 TI - The choice of HIV protease inhibitor: indinavir is currently the best option. AB - (1) Recent consensus recommendations agree that first-line treatment of HIV infection should consist of a three-drug regimen combining a protease inhibitor and two nucleoside reverse transcriptase inhibitors. Some recommendations specifically advise against using the current formulation of saquinavir, but none express a preference for one of the other three protease inhibitors currently marketed in France (indinavir, nelfinavir and ritonavir). (2) These HIV protease inhibitors have established efficacy on viral load and the CD4+ lymphocyte count. Saquinavir may have lower virological efficacy. (3) The clinical efficacy of three-drug regimens containing indinavir or saquinavir is well demonstrated in patients at an advanced stage of HIV disease. (4) The risk of viral resistance is not currently a factor in choosing a HIV protease inhibitors. (5) Several epidemiological studies have compared the risk of adverse effects on three-drug regimens including indinavir, ritonavir or saquinavir. In these studies saquinavir was the best-tolerated drug and ritonavir the worst-tolerated. (6) Ritonavir interacts with many drugs. The poor bioavailability of the current saquinavir formulation also leads to risk of interactions. (7) Treatment constraints differ from one protease inhibitor to another, and these must be taken into account case by case. (8) The daily cost of treatment is not currently an important factor in choosing among the various preparations. (9) Taking into account efficacy, adverse effects, interactions and treatment constraints, the combination of indinavir with two nucleoside reverse transcriptase inhibitors currently seems to be the best choice for the largest number of patients. (10) If problems of compliance arise, nelfinavir can be an alternative to indinavir. (11) In patients at an advanced stage of HIV disease who comply well with their treatment, saquinavir can also be an alternative to indinavir. PMID- 10848068 TI - Prevention of diarrhoea in young children: abundant experience in developing countries. AB - (1) The prevention of diarrhoea in young children has been the subject of many intervention studies in developing countries, and their combined results identify the most effective measures. (2) While it lasts, maternal breast-feeding reduces the overall mortality and mortality linked to diarrhoea. (3) In vitamin A deficient children, supplementation reduces overall mortality and deaths specifically due to diarrhoea. (4) Maternal education on weaning can improve children's nutritional status and may thereby reduce mortality related to diarrhoea. (5) The promotion of personal and domestic hygiene reduces morbidity due to diarrhoea. (6) Facilitating access to drinking water and sanitation is a very effective way of reducing overall childhood mortality. PMID- 10848069 TI - Shaking the Blues: Blue Cross and Blue Shield of South Carolina tailors its fulfillment program to target better prospects and pad the bottom line. PMID- 10848070 TI - A chicken (nugget?) in every pot: what's at stake in the budget debate. PMID- 10848071 TI - CHIP and Medicaid outreach and enrollment: a hands-on look at marketing and applications. PMID- 10848072 TI - States want insurance coverage for cancer screening. PMID- 10848073 TI - Shocks that save lives. PMID- 10848075 TI - Home medical equipment receipt in a home care appropriate population. AB - OBJECTIVES: This study examines home medical equipment (HME) receipt for 1,040 veterans considered appropriate for home health services. METHODS: HME receipt was monitored for 12 months using the Department of Veterans Affairs' Prosthetics database. RESULTS: Eighty-three percent received at least one item; averaging 7.4 items (SD = 6.8). The most common items included commodes/bath benches (9%), canes/walkers (7%), safety equipment (7%), liquid oxygen (6%), and wheelchairs (6%). Two functional status variables, home care use and race, correctly classified 69% of HME recipients. Logistic regressions were run for specific equipment; c-indices ranged from .64 to .75. Age, race, income, functional status, risk of hospital readmission, and home care use were significant predictors. DISCUSSION: HME accounted for $4.5 billion in sales (16% of total) for medical products in 1996. As the HME market continues to expand, the characteristics of HME recipients are necessary to project future HME needs in a growing, elderly population. PMID- 10848074 TI - The importance of genetic and environmental effects for self-reported health symptoms: a 30-year follow-up considering survival and selection effects. AB - OBJECTIVES: This study delineated the nature of individual differences in self reported health status across a 30-year period. Potential survival and selection effects on mean levels, variances, and genetic and environmental sources of variance were evaluated. METHODS: Self-reported health status in 1963 was evaluated in 5,229 pairs of twins from the Swedish Twin Registry and in a subsample of 351 pairs surviving to the age of 80 years in 1993. Structural equation modeling evaluated genetic and environmental contributions to total variance and change in variance. RESULTS: For men but not women, the genetic and environmental influences on health symptoms differed between survivors and nonsurvivors. Total variance increased, reflecting an increase in environmental variance, across the 30 years for both genders. Genetic variance decreased longitudinally for men. DISCUSSION: The increase in variation from the mid-50s to the mid-80s appears to be due to an accumulation in environmental variation. There are gender differences that deserve further exploration. PMID- 10848076 TI - Primary and secondary control-enhancing strategies: implications for health in later life. AB - OBJECTIVES: The major goal of this article was to assess the link between control enhancing strategies and health in an older population. In particular, the use of primary-control strategies, which involve modifying the environment (e.g., actively persisting) and compensatory secondary-control strategies, which involve modifying the self (e.g., expecting less of oneself) was studied. METHODS: Participants (n = 241) in a large-scale longitudinal study were interviewed to assess their use of strategies and their health. RESULTS: Health (physical and perceived) was found to vary for those using primary- and compensatory secondary control strategies; however, the nature of this variation depended on age. DISCUSSION: The findings may indicate that primary-control strategies have positive health implications for the young-old but that these same strategies become detrimental to health in late life. The findings could further suggest that compensatory secondary-control strategies become increasingly more adaptive in late life. PMID- 10848077 TI - Clients speak: participatory evaluation of a nonconfrontational addictions treatment program for older adults. AB - OBJECTIVES: To describe the important aspects of the Lifestyle Enrichment for Senior Adults (LESA) program from the clients' perspective and why these aspects were important to them. METHODS: The study used a participatory approach. Researchers collaborated with counselors and clients to develop an evaluation that (a) respected the experiences and knowledge of people in the program and (b) provided an accurate assessment of the important aspects of the program from the clients' perspective. One-to-one interviews were conducted with 52 clients. RESULTS: Clients' relationships with counselors, the positive emotions experienced during individual and group counseling, and the program's flexibility were of key importance. Allowing clients to choose their level of involvement and the general acceptance of drinkers also contributed to perceived benefits of the program. DISCUSSION: The benefits of client-centered addictions therapy warrant closer examination. Senior adults' motivation for behavior change may be highly sensitive to the relational aspects of the treatment approach. PMID- 10848079 TI - Medicare competitive pricing: lessons being learned in Phoenix and Kansas City. PMID- 10848078 TI - The relationship between self-rated health and health status among coronary artery patients. AB - OBJECTIVES: This study examined the descriptive relationship of self-rated health (SRH) with various psychosocial measures, sociodemographic variables, coronary artery disease (CAD) diagnostic/clinical measures, and medically abstracted comorbidities. METHODS: The sample was 2,855 individuals from the Mediators of Social Support (MOSS) study who had at least 75% narrowing in more than one vessel, as indicated by a cardiac catheterization. RESULTS: After adjusting for sociodemographic factors, individuals who rated their health as poor/fair had significantly worse performance on all psychosocial measures and were more likely to be female, non-White, and of a lower socioeconomic status than those who rated their health as being good or better. There were few differences on SRH across various diagnostic/clinical measures of health. DISCUSSION: A single item measure of SRH may be useful; the generalizability of the item must be considered. In this sample of CAD patients, SRH was related more to psychosocial factors than to clinical and disease indicators. PMID- 10848080 TI - It's your heart. PMID- 10848081 TI - From pill-counting to patient care: pharmacists' standard of care in negligence law. PMID- 10848082 TI - External counterpulsation for treatment of chronic stable angina pectoris. PMID- 10848083 TI - Hyperbaric oxygen therapy for wound healing--part II. PMID- 10848084 TI - Hyperbaric oxygen therapy for wound healing--part III. PMID- 10848085 TI - Ultrasonography of the heel for diagnosing osteoporosis and selecting patients for pharmacologic treatment. PMID- 10848086 TI - Uterine artery embolization for treatment of symptomatic uterine fibroids. PMID- 10848087 TI - Rejecting the logic of confinement: care relationships and the mentally disabled under tort law. PMID- 10848088 TI - The two-track model of bereavement: overview, retrospect, and prospect. AB - Sustaining meaning in life, maintaining emotional well-being, and balancing attachments to the living and the deceased are features associated with the psychological (and often private) impact of loss. In the Two-Track Model of Bereavement, the loss process is conceptualized along two distinct but interactive axes that attend to these overt and covert aspects of the response to loss. Whereas Track I is concerned with biopsychosocial functioning in the wake of loss, it is Track II that focuses on the bereaved's ongoing emotional attachment and relationship to the deceased. The contributions of the model to theory and research can serve to clarify our thinking about bereavement as a process resonating throughout one's life. Initially, research and clinical findings from bereaved parents are presented to illustrate the Two-Track Model and its contribution to the deepening of our understanding of loss throughout the life cycle. The contributions of the model to clinical practice are then considered for their ability to clarify our thinking and interventions. Two clinical cases illustrate situations where a predominant focus is on one or the other of these tracks. Ultimately, the Two-Track Model of Bereavement's use extends to the organization and clarification of theory, research, and clinical work. PMID- 10848089 TI - Deathbed scene narratives: a construct and linguistic analysis. AB - The present study explored temporal and self/other dimensions of death attitudes among young adults. One hundred seventy-two undergraduate students were randomly assigned to 1 of 4 conditions: writing about one's death (age unspecified), one's death as a young adult, or the death of another (age unspecified), or the death of other as a young adult. Results indicated that the death of another, but not of oneself, was associated with more realistic considerations of death (e.g., pain, negative emotions). In addition, participants who wrote about death at an early age were less likely to describe their own deaths with negative emotion or to discuss physiological aspects of death and were less likely to express romanticized notions of death. Implications of these results for death anxiety research are discussed. PMID- 10848090 TI - Parental abduction and the push for reconciliation following a life-threatening illness: two case studies. AB - Life-threatening illness may prompt attempts at reconciliation, even when there have been significant rifts between family members. Two case studies of families where there has been a parental abduction of a child are used to illustrate some of the issues that may emerge when there have been serious rifts and an illness. These cases are taken from a larger longitudinal study of families where one parent has kidnapped a child and gone into hiding for a significant period of time. In the first case, it is the parent who recovered the missing child who is ill and seeking greater connection between her child and the parent who abducted. In the second case, the children wanted connection with the ill father who abducted them. In both cases, it is possible that such attempts at reconciliation may not have served the children well. Implications for practitioners dealing with abduction or life-threatening illnesses are included. PMID- 10848091 TI - The efficiency of managed care "patient protection" laws: incomplete contracts, bounded rationality, and market failure. PMID- 10848092 TI - Reproduction in not a major life activity: implications for HIV infection as a per se disability under the Americans with Disabilities Act. PMID- 10848093 TI - Not disabled enough: the ADA's "major life activity" definition of disability. AB - Michelle Friedland argues in this note that the Americans with Disabilities Act fails to adequately distinguish between the separate goals of preventing pure discrimination and providing affirmative accommodation. The Act's conflation of these two different objectives, and its reliance on a single definition of disability for both, hinders its effectiveness in improving the status of individuals with disabilities in the employment setting. To illustrate this, she points to the counterintuitive results reached in recent court decisions. Friedland further traces the legislative origins of the Act's definition of disability and the ambiguity it leaves as to Congress's goals for the Act's employment provisions. She posits three possible goals the Act might be designed to achieve and recommends basic reforms for accomplishing each. Her ultimate conclusion is that provisions dealing with accommodation and discrimination need to be divided so that each can have its own definition of disability. In addition, she believes funding mechanisms for providing accommodation should be altered to ameliorate inequalities in burdens borne by employers and to avoid improper incentives to circumvent the Act. PMID- 10848094 TI - 'Premium' plasma: isn't 'regular' blood safe enough? PMID- 10848095 TI - Relief for the Rx blues: long waits at the drugstore? Incomplete advice? High prices? We prescribe this report. PMID- 10848097 TI - Alcohol: the whole truth. Seven half-truths about drinking, exposed. PMID- 10848096 TI - Brewed, bottled, or powdered: testing the health power of tea. PMID- 10848098 TI - Making medicines affordable. A number of states have for years helped older Americans buy needed prescription drugs. PMID- 10848099 TI - Death, life, and uncertainty: allocating the risk of error in the decision to terminate life support. PMID- 10848100 TI - Regulating through information: disclosure laws and American health care. AB - Efforts to reform the American health care system through direct government action have failed repeatedly. Nonetheless, an alternative strategy has emerged from these experiences: requiring insurance organizations and health care providers to disclose information to the public. In this Article, Professor Sage assesses the justifications for this type of regulation and its prospects. In particular, he identifies and analyzes four distinct rationales for disclosure. He finds that the most commonly articulated goal of mandatory disclosure laws- improving the efficiency of private purchasing decisions by giving purchasers complete information about price and quality--is the most complicated operationally. The other justifications--which he respectively terms the agency, performance, and democratic rationales--hold greater promise, but make different, sometimes conflicting assumptions about the sources and uses of information. These insights have implications not only for health care, but also for other regulated practices and industries. PMID- 10848101 TI - Princeton Insurance Co. v. Chunmuang--does the court's result signal a need for legislative reform broadening malpractice insurance coverage for the benefit of victims of physician sexual abuse? PMID- 10848102 TI - Introductory remarks by the Honorable Justice Stewart G. Pollock at Seton Hall University School of Law. AB - This address is based upon a presentation given at Seton Hall University School of Law's Seventh Annual Health Law Symposium on February 12, 1999. PMID- 10848103 TI - Prescription drugs, alternative designs, and the Restatement (Third): preliminary reflections. AB - This Article is based upon a presentation given at Seton Hall University School of Law's Seventh Annual Health Law Symposium on February 12, 1999. PMID- 10848104 TI - The continuing search for proper perspective: whose reasonableness should be at issue in a prescription product design defect analysis? AB - This Article is based on a presentation given at Seton Hall University School of Law's Seventh Annual Health Law Symposium on February 12, 1999. PMID- 10848105 TI - Manufacturers' liability for drugs and medical devices under the Restatement (Third) of Torts: Products Liability. AB - This Article is based on a presentation given at Seton Hall University School of Law's Seventh Annual Health Law Symposium on February 12, 1999. PMID- 10848106 TI - Remarks by the Honorable John E. Keefe at Seton Hall University School of Law. AB - These remarks are based upon a presentation given at Seton Hall University School of Law's Seventh Annual Health Law Symposium on February 12, 1999. PMID- 10848107 TI - Compared to what? Instructing the jury on product defect under the Products Liability Act and the Restatement (Third) of Torts: Products Liability. AB - This Article is based on a presentation given at Seton Hall University School of Law's Seventh Annual Health Law Symposium on February 12, 1999. PMID- 10848108 TI - A "safer mousetrap": the evolution of a model charge for alternative safer design cases. AB - This address is based upon a presentation given at Seton Hall University School of Law's Seventh Annual Health Law Symposium on February 12, 1999. PMID- 10848109 TI - Health care fraud and abuse data collection program: reporting of final adverse actions. Office of Inspector General (OIG), HHS. Final rule. AB - This final rule establishes a new CFR part to implement the statutory requirements of section 1128E of the Social Security Act, as added by section 221(a) of the Health Insurance Portability and Accountability Act (HIPAA) of 1996. Section 221(a) of HIPAA specifically directs the Secretary to establish a national health care fraud and abuse data collection program for the reporting and disclosing of certain final adverse actions taken against health care providers, suppliers and practitioners, and to maintain a data base of final adverse actions taken against health care providers, suppliers and practitioners. PMID- 10848110 TI - Forecasting the future. PMID- 10848111 TI - The ABCs of PBMs. PMID- 10848112 TI - Is the Clean Air Act unconstitutional? PMID- 10848113 TI - Scintimammography (also known as "mammoscintigraphy" or "breast scintigraphy"). PMID- 10848114 TI - Cranial orthosis for plagiocephaly without synostosis. PMID- 10848115 TI - FDG positron emission tomography in colorectal cancer. PMID- 10848116 TI - FDG positron emission tomography in lymphoma. PMID- 10848117 TI - FDG positron emission tomography in melanoma. PMID- 10848118 TI - FDG positron emission tomography in pancreatic cancer. PMID- 10848119 TI - High-dose chemotherapy with autologous stem cell support in the treatment of chronic lymphocytic leukemia or small lymphocytic lymphoma. PMID- 10848120 TI - In utero fetal surgery for sacrococcygeal teratoma. PMID- 10848121 TI - Monitoring of bone density to assess active treatment of osteoporosis. PMID- 10848122 TI - Redefining the right to reproduce: asserting infertility as a disability under the Americans with Disabilities Act. PMID- 10848123 TI - Law making for baby making: an interpretive approach to the determination of legal parentage. AB - This article proposes a new model for analyzing legal issues arising from technological conception and uses it to develop rules to govern the legal parentage of technologically conceived children. Professor Garrison shows that most commentators on technological conception have employed a "top-down" methodology, deriving rules for specific cases from an abstract global principle such as reproductive autonomy, freedom of contract, or anticommodification. Professor Garrison critiques these and several other approaches, showing that they offer little concrete guidance in many cases, risk the introduction of discordant values into the law of parentage, and fail to capture all of the values that have traditionally guided parentage determination. In their place, she proposes an "interpretive methodology" which, by relying heavily on current rules governing parentage determination in other contexts, would assimilate technological conception within the broader law of parental obligation. Professor Garrison argues that cases of sexual and technological conception should be governed by similar rules because, despite mechanical differences between these two reproductive methods, there are no significant differences in the parent-child relationships that they produce. She demonstrates that the interpretive approach can cabin rule-making disagreements, and that it can generate comprehensive parentage rules that are based on uniform policy goals and that ensure consistent treatment of parent-child relationships. PMID- 10848124 TI - Urinary incontinence and risk of death among community-living elderly people: results from the National Survey on Self-Care and Aging. AB - OBJECTIVES: The authors explored whether urinary incontinence (UI) is associated with higher mortality and, if so, whether adjustment for demographics, health, and functional status diminishes the association. METHODS: The Cox proportional hazards model was used with a nationally representative sample of community living elderly people (N = 3,485). RESULTS: In an analysis adjusted for age, gender, education, and probability sampling design, participants with mild to moderate UI (hazard ratio [HR] = 1.51; 95% confidence interval [CI] = 1.09-2.08) and severe UI (HR = 1.54; 95% CI = 1.03-2.29) experienced a higher risk of death. Adjustment for health and functional status diminished the association of UI and risk of death (mild to moderate UI, HR = 1.20, 95% CI = 0.85-1.68; severe UI, HR = 0.91, 95% CI = 0.59-1.39). DISCUSSION: The relationship between UI and mortality in older adults can largely be understood on the basis of increased frailty in incontinent individuals. PMID- 10848125 TI - Quantitative genetic analysis of biobehavioral markers of aging in Swedish studies of adult twins. AB - OBJECTIVES: This article will examine the generalizability of markers of aging across samples and cultures and the genetic and environmental influences on them. METHODS: As part of two studies, 18 demographic, cognitive, physiological, personality, and behavioral variables were available from 459 twin pairs ranging in age from 40 to 90 years. Twelve markers of aging were identified using stepwise regression. Factor analysis of the markers produced four factors: general knowledge, fluid abilities, cardiovascular functioning, and well-being. RESULTS: Analysis of twin similarity for the factors suggested that genetic and environmental influences varied greatly. Significant age differences in heritability were found for three of the four factors. DISCUSSION: Results indicate one aging theory cannot account for changes in all markers of aging. Aging of various systems occurs as a result of different combinations of genetic and environmental influences. PMID- 10848126 TI - The effect of race and health-related factors on naming and memory. The MacArthur Studies of Successful Aging. AB - OBJECTIVES: The purpose of the analyses was to examine the impact of health related variables on race differences in neuropsychological functioning (Boston Naming Task). METHODS: Using cross-sectional data from the MacArthur Successful Aging Study, the authors examined the relationship of demographic characteristics, health status, health habits, physical functioning, and speed of performance to naming and incidental recall of items from the Boston Naming Task. Participants were 1,175 healthy African American and European American older persons 70 to 79 years old. RESULTS: Regression analyses indicated that although race differences persisted for confrontational naming after controlling for demographic and health factors, there was no effect due to race for incidental recall scores or for savings scores for recall. DISCUSSION: The racial differences found in test performance may reflect differences in cultural appropriateness of the material rather than differences in ability. PMID- 10848127 TI - Effects of caregiving, gender, and race on the health, mutuality, and social supports of older couples. AB - OBJECTIVES: The purpose of this article is to report caregiving, gender, and race effects on the physical health, social supports, and mutuality of older couples. METHODS: The study involves cross-sectional results from a longitudinal study of 234 older couples, 118 in caregiving situations and 116 comparison couples. Spouses were interviewed in their homes. RESULTS: Spousal older caregivers are in poorer physical health than noncaregivers but have no greater numbers of social supports to help them in the caregiving role. Also, African Americans are in poorer physical health than Caucasians; yet, as caregivers, they have no greater numbers of social supports than Caucasian caregivers. Although females have more social supports than males, female caregivers have more negative feelings about their husbands than male caregivers have about their wives. DISCUSSION: Males and African Americans are in potentially more severe caregiving situations than their female and Caucasian counterparts. Results of this study are compared to Wallsten (1999), which found that African Americans and males had more favorable psychological indicators of stress. The two studies complement each other but open the question of response bias as a factor. PMID- 10848128 TI - Perceptions of health and use of ambulatory care: differences between Korean and White elderly. AB - This study identifies whether culturally based differences in perceptions of health resulted in differences in ambulatory care use among the elderly. METHODS. The authors conducted stratified Poisson regressions on data from a 1992 survey of older Koreans and Whites in Los Angeles County. The models included measures of demographics, health, functioning, income, insurance, social support, and culture (perceptions of health or other beliefs). RESULTS. Descriptive findings showed older Koreans had more ambulatory physician visits, poorer functioning, and poorer perceptions of their health than Whites. Multivariate findings showed that positive perceptions of health independently reduced office visits for both Koreans and Whites, but the effect was significantly smaller for Koreans. Other cultural differences also affected use. DISCUSSION. The findings highlight differences between older Koreans and Whites' responses to physical and socioeconomic conditions and the importance of cultural sensitivity in the health care delivery system. PMID- 10848129 TI - Physician connectivity: electronic prescribing. PMID- 10848130 TI - Human cloning: is the reach of FDA authority too far a stretch? PMID- 10848131 TI - Stuffed! That lingering 'cold' could be sinusitis. PMID- 10848132 TI - Workers comp: falling down on the job. PMID- 10848133 TI - Are you taking too much medicine? The standard recommended starting dose may not be the best choice for you. PMID- 10848134 TI - Spiritual aspects of loss among partners of men with AIDS: postbereavement follow up. AB - This article is a follow-up study of bereaved caregiving male partners of men with AIDS (T.A. Richards & S. Folkman, 1997). The earlier study examined spiritual beliefs, experiences, and practices reported in interviews with 125 caregivers conducted 2 and 4 weeks following bereavement. This follow-up study reports qualitative and quantitative data from 70 members of the earlier cohort, collected 3 to 4 years later, regarding the presence of spiritual phenomena. Spirituality increased or deepened in 77% of the entire cohort. An ongoing relationship with the deceased partner was reported by 70% of the cohort. The use of spirituality as coping appeared to decline as the bereaved moved further in time from the loss. Instead, spirituality was identified as a personal governing influence that provided value and direction to the individual. In general, relationships between the expression of spirituality and mood, coping, and physical health symptoms were not statistically significant owing to small samples, but there were medium effect sizes. PMID- 10848135 TI - Changes in parents' mental distress after the violent death of an adolescent or young adult child: a longitudinal prospective analysis. AB - This study examined changes in bereaved parents' mental distress following the violent deaths of their 12- to 28-year-old children. A community-based sample of 171 bereaved mothers and 90 fathers was recruited by a review of medical examiner records. Data were collected 4, 12, and 24 months post-death. Repeated measures analysis of variance showed significant reductions in 8 of 10 measures of mental distress among mothers and 4 of 10 for fathers, with the most change for both genders occurring between 4 and 12 months post-death. During the 2nd year of bereavement, mothers' symptoms continued to decline, whereas fathers, who started out with less distress than mothers, reported slight increases in 5 of 10 symptom domains. Nonetheless, 2 years after the deaths, mothers' mental distress scores were up to 5 times higher than those of "typical" U.S. women and fathers' scores were up to 4 times higher than "typical" U.S. men. Of the 7 intervening variables examined, higher scores on self-esteem and self-efficacy predicted lower distress for both mothers and fathers 4, 12, and 24 months post-death. Repressive coping was predictive of distress among fathers. It was concluded that violent death bereavement has sustained, distressing consequences on parents of children who die as a result of accidents, homicides, and suicide. PMID- 10848136 TI - Together in spirit: keeping our relationships alive when loved ones die. AB - The premise of this article is that it can be adaptive to maintain bonds with loved ones who have died. The article describes 5 useful strategies for connecting with the deceased, which include recognizing the ways in which the deceased leave an imprint upon our lives, deriving a sense of meaning from the deceased's life, using objects that symbolically link us to persons who have died, identifying ceremonial opportunities for including the deceased in our lives, and reviewing the deceased's life story. Following discussion of these strategies, implications for counseling and outreach are briefly addressed. Finally, discussion focuses on how acknowledging our bonds with the deceased has the potential to aid us in confronting our own mortality. PMID- 10848137 TI - Advanced lumbar puncture. PMID- 10848138 TI - RNFA. PMID- 10848139 TI - Physical and psychosocial correlates of hormone replacement therapy with chronically stressed postmenopausal women. AB - OBJECTIVES: This study investigated the relationship between hormone replacement therapy (HRT) use and physical and psychosocial functioning in a sample of chronically stressed older women. METHODS: Participants (N = 94) were postmenopausal women caring for a parent or spouse with dementia. Caregivers were evaluated using both psychological and physical health indices. RESULTS: Caregivers receiving HRT reported lower levels of hostility and were less likely to report unpleasant interactions with members of their support network; these findings were unrelated to demographic variables, caregiving-related indices, or health behaviors. HRT recipients had lower 10-year risk scores for cardiovascular disease, but there were no group differences in stroke risk. The largest effects on hostility, depression, heart and stroke risk were observed in caregivers receiving a combination of estradiol and progesterone. DISCUSSION: HRT for postmenopausal women may promote positive outcomes for some, but not for all, affective states and cardiovascular variables vulnerable to chronic life stressors. PMID- 10848140 TI - Postacute care following stroke or hip fracture: single services and combinations used by Medicare beneficiaries (1987-1992). AB - OBJECTIVE: To describe the use of postacute services alone or in combination following a hospitalization for a hip fracture or stroke by Medicare beneficiaries who were relatively well and living in the community prior to the index event. METHODS: Health-service use histories were constructed using Medicare claims. Patients in the study represented all subjects from a 1% sample of Medicare beneficiaries who were age 70 years or older at the time of the index hospitalization. RESULTS: From 1987 to 1992, the proportion of patients receiving any postacute care and those receiving combinations of care increased. For example, there was a doubling of the proportion of patients with either condition using sequences of rehabilitation with home health or SNF and home health. Within 1 year of the hospitalization, 42.6% of patients with stroke and 35.0% post-hip fracture had been rehospitalized. DISCUSSION: Resource use and assessment of patient outcomes should be examined across the continuum of postacute care and in the months beyond to examine the relative effectiveness of different combinations of care. PMID- 10848141 TI - A Danish population-based twin study on general health in the elderly. AB - OBJECTIVES: To study the relative influence of genetic and environmental factors on self-rated health and hospitalization patterns in the elderly. METHODS: A survey among all 3,099 Danish twins ages 75 years and older identified in the Danish Twin Registry. An interview was conducted with 77% of the twins. The number of hospitalizations in the previous 18 years was obtained through register linkage, thereby obtaining health information on 96% of the study population, including all nonresponders. RESULTS: Structural equation modeling suggested that approximately a quarter of the variation in the liability to self-reported health and the number of hospitalizations could be attributed to genetic factors. The remaining variation was most likely due to nonfamilial environment. Analyses of the hospitalization patterns of proxy responders and nonresponders suggest that the estimates of the genetic influence on health outcomes in the study are conservative. DISCUSSION: The present study indicates that variation in general health among the elderly is partly explained by genetic factors. PMID- 10848142 TI - Nonfatal agricultural injuries among Colorado older male farmers. AB - OBJECTIVES: This study described characteristics and potential risk factors for nonfatal agricultural injuries among older male farmers age 60 years and older in Colorado. METHODS: A telephone survey was conducted among 113 older male farmers between 1993 and 1995. Agricultural injuries and potential risk factors were evaluated. RESULTS: A total of 27 injuries were sustained. The leading external cause of injuries was livestock, and most of the injuries affected lower and upper extremities of the body. Injured farmers were more likely to be persons who had gone deeply into debt or who were using prescription medications. DISCUSSION: Older farmers are at risk of injuries. The study results support the need to consider physical limitations of aging in developing prevention programs in agriculture. PMID- 10848143 TI - Intervention effects on dementia caregiving interaction: a stress-adaptation modeling approach. AB - OBJECTIVES: Our purpose was to evaluate the adequacy of a stress adaptation framework for guiding intervention research on caregivers and patients coping with Alzheimer's disease, and to test the effect of a cognitive stimulation intervention as an interactive outcome. METHODS: Using a repeated measures design, 87 caregiver-patient dyads were randomized to one of three conditions: active cognitive stimulation, passive stimulation, or control. Assessments occurred at preintervention, postintervention (3 months), and 9 months. RESULTS: The LISREL model was entirely satisfactory by the chi-square goodness-of-fit criteria. However, the coefficients associated with the paths between the mediating concepts and the dyadic interaction differed significantly at 3 months and 9 months. The intervention group caregivers were shown to be more satisfied with their interaction with the impaired member. DISCUSSION: The improvement in caregiver satisfaction was attributed to an attenuation of the behavioral stressor effects through increased use of a problem-focused coping strategy, namely, positive reappraisal of the stressful situation. PMID- 10848144 TI - Clinical experience with Goal Attainment Scaling in geriatric care. AB - OBJECTIVES: This article presents a qualitative evaluation of the utility of Goal Attainment Scaling (GAS) in geriatric care. GAS is an individualized outcome measure particularly suited for clients with multiple complex problems, such as are commonly served by geriatric programs. METHODS: Semistructured interviews were completed with 24 clinicians in four hospital or community-based geriatric services where GAS has been used. An inductive analysis was used to identify themes. RESULTS: GAS was described as resulting in shorter lengths of stay, more focused care, and improved team functioning. GAS was perceived as having greater use for functional (versus medical) goals, when clinicians had more direct control over treatments, when patients were involved in goal-setting, and when goals were set by an interdisciplinary team. DISCUSSION: GAS has affected both care practices and patient outcomes. This study illustrates the clinical impacts an outcome measure can have, as well as the use of qualitative methods for this type of research. PMID- 10848145 TI - Immune globulin and the shortage of plasma derivatives. PMID- 10848146 TI - TEC findings on the use of IVIg. PMID- 10848147 TI - Sentinel lymph node biopsy for breast cancer. PMID- 10848148 TI - Legal drugs? Not without legal reform: the impact of drug legalization on employers under current theories of enterprise liability. PMID- 10848149 TI - The State Attorney Generals' tobacco suits: equitable remedies. PMID- 10848150 TI - Health care reform for the twenty-first century: the need for a federal and state partnership. PMID- 10848151 TI - The dual process model of coping with bereavement: rationale and description. AB - There are shortcomings in traditional theorizing about effective ways of coping with bereavement, most notably, with respect to the so-called "grief work hypothesis." Criticisms include imprecise definition, failure to represent dynamic processing that is characteristic of grieving, lack of empirical evidence and validation across cultures and historical periods, and a limited focus on intrapersonal processes and on health outcomes. Therefore, a revised model of coping with bereavement, the dual process model, is proposed. This model identifies two types of stressors, loss- and restoration-oriented, and a dynamic, regulatory coping process of oscillation, whereby the grieving individual at times confronts, at other times avoids, the different tasks of grieving. This model proposes that adaptive coping is composed of confrontation--avoidance of loss and restoration stressors. It also argues the need for dosage of grieving, that is, the need to take respite from dealing with either of these stressors, as an integral part of adaptive coping. Empirical research to support this conceptualization is discussed, and the model's relevance to the examination of complicated grief, analysis of subgroup phenomena, as well as interpersonal coping processes, is described. PMID- 10848152 TI - Barriers to accessing hospice services before a late terminal stage. AB - Although hospice care has been available in the United States for over 25 years, the trend of late terminal access persists. Quality assessments and interventions that will continue through a year of bereavement are based on early interactions and the formation of caring and trusting bonds. A qualitative research study was conducted to identify and describe barriers to timely access as perceived by those involved in hospice care: caregivers, hospice staff and volunteers, and referring physicians. Major findings include persistent knowledge deficits including misconceptions and lack of awareness of components and skill associated with hospice care: reluctance to disclose, discuss, or acknowledge patient terminality, and access of hospice care being perceived as giving up on the patient. PMID- 10848153 TI - The role of the mental health professional in helping significant others of persons who are assisted in death. AB - The involvement of mental health professionals with people who are considering assisted death due to a terminal illness is beginning to be discussed more frequently in the professional literature; however, one neglected area is working with the significant others of the person who is suffering. This article covers ways both before and after the death that counselors can help significant others cope with the dying process, the decision to request assisted death, and the death itself. PMID- 10848155 TI - The effect of disabilities on exits from AFDC. AB - The Personal Responsibility and Work Opportunity Reconciliation Act (PRWORA) of 1996 replaces AFDC, the largest means-tested cast assistance program for low income families, with the Temporary Assistance for Needy Families (TANF) block grant. Unlike AFDC, assistance under TANF is limited to five years in a lifetime, and states are required to move families from the assistance rolls into jobs. But not all adult welfare recipients can easily move to work because either they themselves are disabled or they have a child with disabilities requiring special care. This article examines the extent and impact of disability among families on AFDC to gain insight into the potential impact of changes under TANF. Using data from the 1990 Survey of Income and Program Participation (SIPP), we find that in nearly 30 percent of the families on AFDC either the mother or child has a disability. Furthermore, we find that having a disability significantly lowers the probability that a woman leaves AFDC for work but not for other reasons, such as a change in living arrangements. Finally, we find little evidence that having a child with a disability affects the probability of leaving AFDC for any reason. PMID- 10848154 TI - The relationship of attachment theory and perinatal loss. AB - Perinatal loss has recently received attention in the literature that presents it as different from other types of losses. Perinatal loss, or the loss of an infant due to miscarriage, stillbirth, or neonatal death, is a significant problem that may be best understood when viewed through the framework of attachment theory. Recent advances in medical technology, including prenatal diagnostic procedures and resulting decisions have influenced issues of both perinatal attachment and loss, and have provided challenges for the clinician. This article presents a review of current theories and research on attachment and perinatal loss, and discusses how knowledge gained from this research may be integrated into clinical practice. PMID- 10848156 TI - Financial incentives for childhood immunization. PMID- 10848157 TI - Synthetic risks, risk potency, and carcinogen regulation. AB - This article analyzes a comprehensive sample of over 350 chemicals tested for carcinogenicity to assess the determinants of the probability of regulation. Controlling for differences in the risk potency and noncancer risks, synthetic chemicals have a significantly higher probability of regulation overall: this is due to the greater likelihood of U.S. Food and Drug Administration (FDA) regulation. Measures of risk potency increase the probability of regulation by the U.S. Environmental Protection Agency (EPA), have a somewhat weaker positive effect on regulation by the U.S. Occupational Safety and Health Administration (OSHA), and decrease the likelihood of regulation by the FDA. The overall regulatory pattern is one in which the FDA targets synthetic chemicals and chemicals that pose relatively minor cancer risk. The EPA particularly performed more sensibly than many critics have suggested. PMID- 10848158 TI - Assessing alternative drug control regimes. AB - The debate over alternative regimes for currently illicit psychoactive substances focuses on polar alternatives: harsh prohibition and sweeping legalization. This study presents an array of alternatives that lies between these extremes. The current debate lacks an explicit and inclusive framework for making comparative judgments. In this study, we sketch out such a framework, as a reminder of possible policy levers and their costs and benefits that might otherwise be neglected or go unrecognized. The framework identifies a range of pharmacological and economic characteristics of substances, potential harms and their bearers, and the sources of those harms, including drug use, trafficking, law enforcement, and illegal status per se. The framework highlights the difficulty of making objective, rigorous comparisons among regimes, but we believe that it can serve a useful heuristic role in promoting more constructive debate and identifying fruitful questions for research. PMID- 10848159 TI - Abandonment of residential housing and the abatement of lead-based paint hazards. PMID- 10848160 TI - Assessing Medicaid managed care in eastern state. AB - This case examines the expanding role of managed care programs in improving health care for the poor while controlling runaway health care costs. The case asks what the commissioner of health in a large eastern state should do to effectively monitor Medicaid managed care programs in her state. The commissioner faces intense pressures for cost containment and strong, but not universal, support for the managed care solution to health care cost problems. The commissioner is herself concerned that the cost savings attributed to managed care may not be real and that the unintended effects on health care may be adverse. Her immediate challenge is to determine what kinds of data she should require service providers to submit to her agency so that she may effectively monitor managed care programs for health care quality, provide positive feedback to health care providers, and establish politically credible program oversight. PMID- 10848161 TI - Ambulatory blood pressure monitoring for diagnosing hypertension in patients with elevated office blood pressure. PMID- 10848162 TI - Apolipoprotein E epsilon 4 allele (APOE epsilon 4) and Alzheimer's disease: role of genetic testing for diagnosis and risk assessment. PMID- 10848163 TI - Becaplermin for wound healing. PMID- 10848164 TI - Intracoronary radiation to prevent restenosis after percutaneous transluminal coronary angioplasty with and without stent placement. PMID- 10848165 TI - Lipid apheresis in the treatment of patients with severe, refractory hypercholesterolemia. PMID- 10848166 TI - Lung volume reduction surgery for severe emphysema. PMID- 10848167 TI - Off-label uses of trastuzumab. PMID- 10848168 TI - Phototherapy for the treatment of seasonal affective disorder. PMID- 10848169 TI - Total purchasing in primary care: three case studies. AB - Total purchasing in primary care (TPPC) was piloted in the mid-1990's, first by four "pioneer sites", and later by over 80 first and second waves of "pilot sites". Separate evaluations of three of the pioneers show that they faced challenges similar to those experienced by the pilot sites, namely: the need to develop organisations which were effective both internally and, in their relationships with other health-care agencies, externally; and the difficulties encountered when attempting to change the behaviour of local acute care providers. Although GP fund holding and/or TPPC have had some successes in influencing the pattern of delivery for some elective and community services, the challenge of reducing hospital admissions and lengths of stay remains formidable. PMID- 10848170 TI - Sources of career disadvantage in nursing. A study of NHS Wales. AB - Despite the numerical predominance of women in nursing there is a marked concentration of women, especially those working part-time, in the lower echelons of the profession. The paper presents survey data and interview material from a study of qualified nurses in NHS Wales. By controlling for differences in education and experience in nursing work, it was found that comparable groups of female nurses received unequal employment opportunities. Women with dependent children were primarily located in the lower nurse grades irrespective of their qualifications and experience. Much of this was associated with inflexible working practices, and the low status of part-time work. Occupational downgrading for female returners was also a significant barrier to career advancement. However, these problems were not recognised by management. Management failed to evaluate the mechanics of their human resource policies in terms which matter to many nurses, in particular with regard to the management of diversity. PMID- 10848171 TI - Integrated care management: applying control theory to networks. AB - Delivery of integrated care by interorganizational networks attracts much attention in Europe. Such care is required to meet the demands of multi-problem patients. Many efforts are made to establish networks. Often, established networks do not deliver integrated care. Managers must understand the background of this problem, in order to deal with it. The issue addressed here concerns behaviour control in networks of autonomous care-providing organizations. So far, publications have focused on behaviour control in single organisations. Based on empirical data we argue that, due to an essential distinction between networks and single organizations, behaviour control in the former should be approached differently. In addition, we discuss the implications of our findings for the management of integrated care delivery. PMID- 10848172 TI - Limits to employee empowerment in the UK NHS. Locality managers in a community health trust. AB - The concern of this paper lies with empowerment of middle managers in a community health trust. It considers the impact of traditional structures and cultures upon the level of decision which middle managers are empowered to make. The conclusion reached is one which suggests that the level of discretion available to middle managers was one of making task-oriented decisions rather than decisions about strategic change. However, even this was compromised by financial constraints and by "best practices" imposed by the personnel function. PMID- 10848173 TI - Identifying the management development needs of senior executives in Iran's teaching hospitals. AB - This research paper reports the findings of the first comprehensive survey of senior executives in Iran's teaching hospitals. It is based on an analysis identifying the continuing professional development (CPD) needs of the total population of the two senior levels of teaching hospitals management-presidents of physician-managers and administrative-managers. Four key areas of management knowledge were selected as the focus of the need identification: operational; financial; human resource; and organization change. The findings reveal a pent up demand for introductory level, formal knowledge in all four areas of management theory, to complement and extend the practical experience they have acquired in managing the complex environment of teaching hospitals. The paper goes on to propose a curriculum design and overall framework of provision to meet these genuinely felt CPD needs. PMID- 10848175 TI - Spotlight. Interview by Sarah Powell. AB - Professor Brain Edwards, Professor of Health Care Development at the University of Sheffield, UK talks to Sarah Powell about changes in emphasis in health-care service management, inequalities of access to health care, the concept of mutuality, and the challenges of the future. PMID- 10848174 TI - Best practice organizational effectiveness in NHS Trusts. Leeds Teaching Hospitals NHS Trust. Case study. AB - Measuring organisational effectiveness in a health-care delivery context is quite a challenging task. Although there are numerous performance assessment models, audit tools and managerial diagnostic tools, they all, however, tend to fall short in their attempts to scrutinize how health-care organizations deploy their capabilities to deliver optimum quality in service provision and what performance levels they achieved as a result of their approach. The project reported here attempted to address these issues, reflecting the experience of Leeds Teaching Hospitals, one of a series of Trusts whose approach to organizational effectiveness was closely examined. PMID- 10848176 TI - Clinical governance: principles into practice. AB - This paper describes the early development of clinical governance in an acute NHS Trust. Three Trust-wide workshops and 17 individual departmental workshops were held between 1998-1999. The discussions in these workshops were used to define the key founding principles of clinical governance and the operational structure. The philosophy behind clinical governance, to improve quality of services, was recognised as being part of mainstream trust business, not an optional add-on. The authors found that teamwork and multidisciplinary collaboration are essential components of future quality improvement. Effective leadership skills need to be supported and developed, with responsibilities shared between a core group within each department rather than one individual. Contributions should be recognised and rewarded. Collaboration with primary care and involvement of patients are prerequisites. Specific objectives should be agreed by each department and used to monitor progress. More effective use of existing resources (staff, time, IT and training) can be made. PMID- 10848177 TI - A devastated continent. AIDS has only begun to take its deadly toll on southern Africa. PMID- 10848179 TI - A cure that may cost us ourselves. Society: one of the pioneers of human genetic engineering predicts that within 30 years, there will be a gene-based therapy for most diseases. But he fears the profound dangers of his own work. PMID- 10848178 TI - Rewiring your gray matter. The brain: you can teach an old brain new tricks. Neuroplasticity promises to give a whole new meaning to 'changing your mind'. PMID- 10848180 TI - A pig may someday save your life. Biotech: scientists are racing to turn oinkers into organ donors. The effort could bring huge benefits, but it carries huge risks. PMID- 10848181 TI - The war on disease goes miniature. Nanomedicine: drugs and cancer tests, cell by cell. PMID- 10848182 TI - Flirting with strange ideas. Mbeki reaches out to a controversial AIDS researcher. PMID- 10848183 TI - Alzheimer's. Unlocking the mystery. The longer we live, the more likely we are to contract this devastating disease. But recent discoveries are bringing scientists closer than ever to a cure. PMID- 10848184 TI - Coping with the darkness. Revolutionary new approaches in providing care are helping people with Alzheimer's stay active and feel productive. PMID- 10848185 TI - My father's battle with Alzheimer's. You start with a 600-acre ranch and wind up in the den. What I've learned from his struggle. PMID- 10848186 TI - The U.S. Medicaid reform: the current state and policy implications for low income population. AB - Medicaid is the health care program that is financed jointly by the federal and state governments. Many states are seeking ways to contain the increased budgetary strain that has resulted from the increases in Medicaid spending. For many states, managed care has been viewed as the means to hold down costs for some of the population served by Medicaid. This article examines the origins and status of Medicaid and the options available to extend coverage to the low-income population. PMID- 10848187 TI - Screening workers for genetic hypersusceptibility: potential ethical, legal, and social implications from the Human Genome Project. AB - One of the potential outcomes of the Human Genome Project will be the ability to identify individuals who are at increased risk of adverse health effects following exposure to hazardous substances in the workplace because of genetic hypersusceptibility. The ability to identify such individuals is likely to lead to the inclusion of genetic screening in worker protection programs. This technology and its applications will have a number of potential ethical, legal, and social implications. In this commentary, the authors examine five broad topics relating to the use of screening for genetic hypersusceptibility in the workplace: (1) issues of risk; (2) the rationale and legal basis for screening; (3) the privacy concerns of workers; (4) the confidentiality of test results; and (5) potential discrimination. The authors close by suggesting some guidelines for developing policies regarding genetic screening. PMID- 10848188 TI - Phases of burnout among Irish professionals. PMID- 10848189 TI - Burnout in Taipei municipal government. PMID- 10848190 TI - A minimalist test of the phase model: pattern survivability in a small Israeli population. PMID- 10848191 TI - Perspectives on psychological burnout, VII, Part 4: Replications in overseas populations--a symposium. PMID- 10848192 TI - Burnout in Malaysia: a note. PMID- 10848193 TI - Burnout in New Zealand: as good as gold. PMID- 10848194 TI - Phases of burnout in Belize. PMID- 10848195 TI - Clinical capital equipment acquisition: decision-making at the executive level. AB - Capital investment in the United States health care industry is one of the components of the system that has historically fueled cost increases and complicated the quality/cost/access conundrum. This article is concerned with capital development focused on clinical equipment acquisition in acute care hospitals. The research findings suggest that decision-makers are abandoning two approaches to decision-making known as Quantitative Decision-Making and Mixed Scanning in favor of two models known as Rational Decision-Making and Political Decision-Making. While this is the case, a method of decision-making known as Idea Sets (developed and labeled by James March as Garbage Can Decision-Making) is the most widely used approach. Equipment acquisition criteria used by the decision-makers are shifting away from concerns of enhancing existing clinical programs and/or adding new clinical programs. Acquisition criteria are shifting toward procuring equipment which will decrease institutional expense, improve organizational efficiency, and galvanize operational effectiveness. In addition to the technical findings of the research, further insights about ourselves and our colleagues are gleaned. Accordingly, we understand more completely the human dynamics surrounding decisions and thus are able to dialogue more richly about issues. Meaningful dialogue such as this will have the dual benefit of advancing teamwork and facilitating decision-making. PMID- 10848196 TI - Phases of burnout among Israeli nurses. PMID- 10848197 TI - Using relationship marketing to develop and sustain nurse loyalty: a case of a rural health care institution. AB - The prosperity of a health care organization is contingent on its ability to compete for and retain a high quality staff of "loyal" nurses. Although the benefits of maintaining a loyal nursing staff are obvious, turnover in the health care industry is dangerously high. One solution for reducing turnover is to develop and sustain a loyal nursing staff. The purpose of this article is to apply customer-oriented marketing theories and practices to better understand how strong nurse-provider relationships can be developed and maintained over time. The authors first examine relationship marketing literature as it applies to nurse relationship and management issues. Second, a framework for conceptualizing internal marketing efforts devoted to enhancing nursing staff satisfaction and retention in tested. Finally, strategies for practicing relationship marketing will be provided. PMID- 10848198 TI - I learned my bedside manner the hard way. PMID- 10848199 TI - Don't get stuck by thorny family situations. PMID- 10848200 TI - Hospital peer review is a kangaroo court. PMID- 10848201 TI - The word I couldn't bring myself to use. PMID- 10848202 TI - Okay, doc, let's talk price. PMID- 10848203 TI - Why we need code auditing. PMID- 10848205 TI - Get a medical license? It's easier to get a gun. PMID- 10848204 TI - Your patient thought you said what? PMID- 10848206 TI - Should these doctors have let their colleague die? PMID- 10848207 TI - From specialist to generalist: nothing comes easy. PMID- 10848208 TI - How many of your patients have returned from the dead? PMID- 10848209 TI - Biotech battle royale. Two firms fight over billion-dollar drug. PMID- 10848210 TI - The cells of immortality. Scientists are tinkering with natural limits on life span. Is that a good idea? PMID- 10848211 TI - A pain in the neck, and then some. The debate over an ergonomics standard. PMID- 10848212 TI - Mimicking mom's milk. A formula for smarts. PMID- 10848213 TI - Bibliography. Current world literature. Glaucoma. PMID- 10848214 TI - Heredity in primary open-angle glaucoma. AB - The past years have seen considerable progress in the characterization of hereditary factors in primary open-angle glaucoma. Epidemiologic studies strengthened our knowledge of the hereditary factors in this multifactorial disease. Several loci in the human genome have been described, which segregate with different glaucoma phenotypes. Mutations of the MYOC/TIGR (myocilin/trabecular meshwork inducible glucocorticoid response) gene on chromosome 1q account for most, but probably not all, cases of glaucoma linked to chromosome 1q, and other additional pathologic factors may be implicated. The properties of the normal myocilin protein point to a crucial role in the regulation of intraocular pressure. However, in spite of the knowledge obtained so far, routinely performed genetic screening of patients at risk for primary open-angle glaucoma is not yet clinically useful. PMID- 10848215 TI - Neuroprotection: a new treatment modality for glaucoma? AB - It is now commonly accepted that glaucoma is a neurodegenerative disease of the optic nerve. Thus, at any given time, there are neurons that, though still viable, are vulnerable to the hostile extracellular milieu and are therefore amenable to neuroprotective therapy. Neuroprotection refers to any intervention, either external to the optic nerve or internally, that will lead to an intracellular change in the balance between survival and death signals in favor of survival. Several potential sites and modalities for such intervention may exist. When designing neuroprotective therapy, ways must be sought to recruit the physiologic self-repair mechanisms awakened by the primary or secondary risk factors. These mechanisms appear to be insufficiently effective when in their natural state, but they may be simulated or boosted by appropriate therapeutic compounds or cells. PMID- 10848216 TI - The mechanism of action of prostaglandins on uveoscleral outflow. AB - It is generally accepted that prostaglandins (PGs) lower intraocular pressure by increasing uveoscleral outflow. The growing use of PGs to lower intraocular pressure has led to increased interest in the uveoscleral outflow. Uveoscleral outflow passes through extracellular spaces within the ciliary muscle and then through the suprachoroidal space to the posterior pole of the eye. Recent studies indicate that this reflects a direct effect of PGs on specific ciliary muscle prostanoid receptors. Activation of these receptors stimulates several linked responses, including cAMP formation and induction of c-Fos and c-Jun expression. These signals lead to increased biosynthesis of matrix metalloproteinases, a family of neutral proteinases that can cleave extracellular matrix molecules. These matrix metalloproteinases may initiate the alteration of collagens in the ciliary muscle to increase spaces among ciliary muscle fibers, thereby reducing hydraulic resistance in the uveoscleral outflow pathway. PMID- 10848217 TI - The optic nerve head in normal-tension glaucoma. AB - There is controversy over the definition, appearance, and characteristics of the optic nerve head in normal-tension glaucoma (NTG). Optic disk size is greater in eyes with NTG than in those with primary open-angle glaucoma. However, in an intraindividual bilateral comparison, the eye with the larger optic disk showed neither more marked nor less pronounced glaucomatous optic nerve damage. Optic disk hemorrhage and peripapillary atrophy have been reported to be more frequent in patients with NTG. Nonuse of calcium channel blockers, peripapillary atrophy, and disk hemorrhage were statistically significantly associated with visual field loss progression in NTG. However, there is a possibility that a high IOP may stop disk hemorrhage relatively early. Histopathologic optic nerve head changes correlated with the clinical appearance of the optic nerve head, which is comparable in NTG and primary open-angle glaucoma. However, as novel findings, serum antibodies to retinal proteins and retinal immunoglobulin deposition in the ganglion cells were observed, and the level of serum autoantibodies to optic nerve head glycosaminoglycans was higher in patients with NTG than in patients with primary open-angle glaucoma. PMID- 10848218 TI - Wound healing modulation after glaucoma surgery. AB - Recent developments in molecular and cell biology have made a major impact on our understanding of the wound healing process and its modification. In this article, the spectrum of therapies that are either currently available or have potential application as modulators of the scarring response following glaucoma surgery are reviewed. Refinement of existing antimetabolite regimens can improve surgical results, especially in carefully selected patient groups. However, the most promising new approach appears to be using molecular-based therapies, such as fully human neutralizing monoclonal antibodies, designed to target specific molecules in the scarring response. Such strategies ultimately offer the potential of safer, more specific, focal, and titratable treatment, with far reaching clinical applications. PMID- 10848219 TI - Endoscopic surgery in glaucoma management. AB - Endoscopically-controlled intraocular surgery has been evolving rapidly since the introduction of thinner endoprobes with increasing resolving power and higher image quality. Currently, anti-glaucomatous surgery is also beginning to take advantage of this exciting new technology. This article discusses the two main approaches to glaucoma management using microendoscopic techniques-decreasing aqueous humor production by means of laser endoscopic cyclophotocoagulation and increasing outflow facility through endoscopically-controlled selective trabecular surgery. Besides presenting clinical results, emphasis is placed on different surgeons' accounts of intraoperative endoscopic maneuverability and the spectrum of complications that may be attributed to the endoscopic nature of the surgical intervention. PMID- 10848220 TI - Quantitative assessment of the anterior segment using ultrasound biomicroscopy. AB - The development of ocular imaging devices has progressed rapidly during the past 10 years. Ultrasound biomicroscopy has revolutionized the evaluation of the anterior segment of the eye. The qualitative information gathered using this technology has contributed to our understanding of the pathophysiology of angle closure glaucoma, pigmentary glaucoma, and a variety of other anterior segment disorders. However, the area of quantitative analysis of ultrasound biomicroscopic images remains largely to be developed. This review describes the role of ultrasound biomicroscopy in the measurement of the anatomic structures and their configurations within the anterior segment. Included are previously published and established methods as well as methods in their early stages of development. Application of quantitative image analysis techniques should yield significant information about mechanisms of appositional angle closure, dynamic functions of the iris, accommodation, and presbyopia. PMID- 10848221 TI - Psychophysical examination of paracentral defects in glaucoma. AB - More than 50% of ganglion cells may be damaged before visual field loss is measurable by conventional methods in primary open angle glaucoma. There is general agreement on the need to improve early diagnosis of visual field loss in primary open angle glaucoma. In this article, new techniques that enlist measurement of paracentral regions are discussed, and the ability of each method to detect visual field loss prior to perimetric loss is described. PMID- 10848222 TI - Is there a role for the immune system in glaucomatous optic neuropathy? AB - Glaucoma and immunity are not traditionally perceived as being causally related. Recently, however, compelling observations have provided insight into a potential role for the immune system in the development of glaucomatous optic neuropathy. In this article, it is proposed that the role of the immune system in glaucoma is two-fold. In some patients, there is evidence that an autoimmune mechanism may be responsible for eliciting damage to the optic nerve, resulting in glaucomatous injury. Autoimmune damage to the optic nerve may occur directly by autoantibodies, or indirectly by way of a "mimicked" autoimmune response to a sensitizing antigen which, in turn, injuries retinal ganglion cells. We suggest that autoimmune-mediated glaucoma injury occurs most often, but not exclusively, in patients in whom the intraocular pressure has never been found to be elevated. A second role of the immune system in glaucoma is likely one of surveillance, in which signal pathways of the immune system regulate cell death in response to conditions that stress retinal neurons in glaucoma. These might include mechanical stress from high intraocular pressure, ischemia, excessive excitatory amino acids, or toxic products resulting from excessive nitric oxide synthase production in either neurons or glial fibers that surround the optic nerve as it exits the eye. In these cases, we propose that the immune system acts as an "arbiter" to help determine whether a neuronal cell will ultimately survive, or succumb to, those stressors that are perceived as injurious. It is conceivable that such surveillance and cell death regulation by the immune system is important in determining the fate of retinal neurons in both the more common "high-pressure" forms of glaucoma, such as primary open-angle glaucoma, and in cases in which the intraocular pressure appears within normal range. PMID- 10848223 TI - Nonpenetrating filtering surgery in glaucoma. AB - Since 1967, trabeculectomy has been the standard drainage operation for most cases of medically uncontrolled glaucoma. It is associated with significant risk of hypotony, hyphema, choroidal effusion or hemorrhage, and bleb failure. Toward the goal of a safer and more reproducible glaucoma filtering surgery, nonperforating filtering surgeries are now performed in primary open-angle glaucoma, normal-pressure glaucoma, and most cases of secondary glaucoma. The site of highest resistance to outflow, the juxtacanalicular trabecular meshwork, is bypassed with avoidance of penetration into the anterior chamber. Different techniques of nonpenetrating filtering surgeries are discussed here. PMID- 10848224 TI - Neuroprotection: is it already applicable to glaucoma therapy? AB - Many categories of both natural and synthetic compounds have been reported to have neuroprotective activity. These include not only antioxidants, N-methyl-D aspartate receptor antagonists, inhibitors of glutamate release, calcium channel blockers, polyamine antagonists, and nitric oxide synthase inhibitors, but cannabinoids, aspirin, melatonin, and vitamin B-12. The lack of availability of specific neuroprotectant compounds in the United States and the lack of clinical trials examining the benefits of neuroprotective agents for glaucoma currently limit the use of these agents. This article provides a short overview of the concept of neuroprotection as it applies to glaucoma and suggests the possibility of neuroprotective activity that might be provided by compounds that are presently easily available. PMID- 10848225 TI - The value of high-pass resolution perimetry in glaucoma. AB - High-pass resolution perimetry (HRP) is a perimetric technique that relies on low contrast, high-spatial-frequency-filtered, ring-shaped test targets. It was devised as a quick and user-friendly technique that reflects the status of the retino-corneal pathway. Since its introduction in 1987, HRP has undergone substantial investigation, primarily in cross-sectional studies on patients with glaucoma, though it is also used in patients with neuro-ophthalmologic disorders. HRP has a shorter test time compared with conventional perimetry. Currently, evidence suggests that HRP is a viable test for the evaluation of the glaucomatous visual field. PMID- 10848226 TI - Collagen degradation in glaucoma: will it gain a therapeutic value? AB - The extracellular matrix is in a constant state of turnover, and several studies suggest that this homeostasis is out of balance in open-angle glaucoma. Recent evidence suggests that matrix metalloproteinases, which are the enzymes primarily responsible for degradation, play a role in numerous modern glaucoma therapies, including topical prostaglandin analogues, topical steroids, and argon laser trabeculoplasty. Additionally, direct and indirect regulation of this system has been shown to increase aqueous humor outflow facility. It is possible that therapies directed at modulating specific enzymes represent the next generation of glaucoma therapy. PMID- 10848227 TI - Latanoprost and cystoid macular edema: is there a causal relation? AB - Published reports of the occurrence of cystoid macular edema (CME) in eyes being treated with latanoprost have led to concern regarding a possible causal relation between the two. Review of all published cases (28 eyes in 25 patients), plus another case reported here for the first time, indicates that all eyes had independent risk for development of CME, so that definitive conclusions about a causal relation cannot be established. In addition, controlled clinical trials and experimental studies with latanoprost have given no indication that latanoprost causes clinical CME. Pharmacokinetic considerations indicate that the concentration of latanoprost expected in the posterior segment of the eye is too low to have a pharmacologic effect, and latanoprost is not known to exhibit vasoactive or inflammatory properties. Nevertheless, reports of a possible association between CME and latanoprost use must be given serious consideration, and in eyes that are at risk for CME, an increased level of surveillance for its development is recommended. PMID- 10848361 TI - Exploring the cost of business. PMID- 10848362 TI - Cost analysis of excimer laser coronary angioplasty versus rotational atherectomy. PMID- 10848363 TI - LVADs--a new era in patient care. AB - Clearly, a challenge exists in treating end stage congestive heart failure patients. Drug therapy is not effective and too few donor hearts prevent transplantation from being the therapy of choice. The only alternative that is developing is the use of LVAD technology. Patients can benefit with an improved quality of life as well as increased longevity. Physicians can benefit by providing their patients with an effective alternative with the potential of reversing the disease process. Costs are significant; however they are comparable in comparison to other treatment arms. With increased use, costs will reduce as the result of developing the efficient use of this technology. PMID- 10848364 TI - Policing the electronic frontier: an introduction to E-health legal issues. PMID- 10848365 TI - Duties of a doctor: UK doctors and good medical practice. AB - OBJECTIVE: To assess the responses of UK doctors to the General Medical Council's (GMC) Good Medical Practice and the Duties of a Doctor, and to the GMC's performance procedures for which they provide the professional underpinning. DESIGN: Questionnaire study of a representative sample of UK doctors. SUBJECTS: 794 UK doctors, stratified by year of qualification, sex, place of qualification (UK v non-UK), and type of practice (hospital v general practice) of whom 591/759 (78%) replied to the questionnaire (35 undelivered). MAIN OUTCOME MEASURES: A specially written questionnaire asking about awareness of Good Medical Practice, agreement with Duties of a Doctor, amount heard about the performance procedures, changes in own practice, awareness of cases perhaps requiring performance procedures, and attitudes to the performance procedures. Background measures of stress (General Health Questionnaire, GHQ-12), burnout, responses to uncertainty, and social desirability. RESULTS: Most doctors were aware of Good Medical Practice, had heard the performance procedures being discussed or had received information about them, and agreed with the stated duties of a doctor, although some items to do with doctor-patient communication and attitudes were more controversial. Nearly half of the doctors had made or were contemplating some change in their practice because of the performance procedures; a third of doctors had come across a case in the previous two years in their own professional practice that they thought might merit the performance procedures. Attitudes towards the performance procedures were variable. On the positive side, 60% or more of doctors saw them as reassuring the general public, making it necessary for doctors to report deficient performance in their colleagues, did not think they would impair morale, were not principally window dressing, and were not only appropriate for problems of technical competence. On the negative side, 60% or more of doctors thought the performance procedures were not well understood by most doctors, were a reason for more defensive practice, and could not be used for problems of attitude. Few differences were found among older and younger doctors, hospital doctors, or general practitioners, or UK and non-UK graduates, although some differences were present. CONCLUSIONS: Most doctors working in the UK are aware of Good Medical Practice and the performance procedures, and are in broad sympathy with Duties of a Doctor. Many attitudes expressed by doctors are not positive, however, and provide areas where the GMC in particular may wish to encourage further discussion and awareness. The present results provide a good baseline for assessing changes as the performance procedures become active and cases come before the GMC over the next few years. PMID- 10848366 TI - Improving surgical care: looking beyond individual competence. PMID- 10848368 TI - Monitoring quality of audit in obstetrics and gynaecology. AB - OBJECTIVE: To develop a questionnaire to assess audit activity and to use it to evaluate systematically the quality of audit in obstetrics and gynaecology within NHS hospitals in the UK. DESIGN: Retrospective review of 212 consecutive questionnaires completed at hospital recognition committee visits for training accreditation, between 1 January 1993 and 31 August 1998, validated against hospital trust annual audit reports. MAIN MEASURES: Use of seven quality criteria developed within the Royal College of Obstetricians and Gynaecologists clinical audit unit and also assessment of support for audit and participation in regional and national audit. Results were compared between 1993/4 (n = 72), 1995/6 (n = 72), and 1997/8 (n = 68) for evidence of improvement. RESULTS: After modifications to the questionnaire the version used from 1993 proved to be a satisfactory tool with minimal need for subsequent change. The results showed that there has been a significant improvement in the quality of obstetric and gynaecology audit with time (p < 0.0001) with 36 (53%) of departments in the previous two year period meeting all seven criteria. Similarly by this stage, 60 (88%) of departments had reached the stage of re-audit and 55 (81%) had conducted patient satisfaction surveys, both of these having significantly improved with time. Critical incident monitoring also became used more widely with time. Validation of topics audited was possible for 45% of hospitals where trust annual audit reports were available and these showed a high level of correlation. CONCLUSIONS: It has proved possible to conduct an audit of audit using the current system of hospital recognition visits for training accreditation. This has shown a great variety in the depth and breadth of audit that is being undertaken within individual obstetric and gynaecology departments. Since 1993 there has been an improvement in the quality of audit programmes undertaken, in particular in the number of hospitals carrying out critical incident monitoring, patient satisfaction surveys, and re-audit. This should be associated with improvements in staff training and in patient care. PMID- 10848367 TI - Reviewing audit: barriers and facilitating factors for effective clinical audit. AB - OBJECTIVE: To review the literature on the benefits and disadvantages of clinical and medical audit, and to assess the main facilitators and barriers to conducting the audit process. DESIGN: A comprehensive literature review was undertaken through a thorough review of Medline and CINAHL databases using the keywords of "audit", "audit of audits", and "evaluation of audits" and a handsearch of the indexes of relevant journals for key papers. RESULTS: Findings from 93 publications were reviewed. These ranged from single case studies of individual audit projects through retrospective reviews of departmental audit programmes to studies of interface projects between primary and secondary care. The studies reviewed incorporated the experiences of a wide variety of clinicians, from medical consultants to professionals allied to medicine and from those involved in unidisciplinary and multidisciplinary ventures. Perceived benefits of audit included improved communication among colleagues and other professional groups, improved patient care, increased professional satisfaction, and better administration. Some disadvantages of audit were perceived as diminished clinical ownership, fear of litigation, hierarchical and territorial suspicions, and professional isolation. The main barriers to clinical audit can be classified under five main headings. These are lack of resources, lack of expertise or advice in project design and analysis, problems between groups and group members, lack of an overall plan for audit, and organisational impediments. Key facilitating factors to audit were also identified: they included modern medical records systems, effective training, dedicated staff, protected time, structured programmes, and a shared dialogue between purchasers and providers. CONCLUSIONS: Clinical audit can be a valuable assistance to any programme which aims to improve the quality of health care and its delivery. Yet without a coherent strategy aimed at nurturing effective audits, valuable opportunities will be lost. Paying careful attention to the professional attitudes highlighted in this review may help audit to deliver on some of its promise. PMID- 10848369 TI - A question sheet to encourage written consultation questions. AB - PROBLEM: Interviews with parents and children attending a hospital paediatric neurology clinic indicated they had difficulties in asking questions during consultations. AIM: To set up a process to enable parents and children to get the information they wanted. BACKGROUND AND SETTING: Two paediatric neurology clinics in separate hospitals in Greater Manchester, UK with a similar client group run by one consultant. DESIGN: Various styles of question sheets were evaluated. The one that was chosen asked patients to write down questions and hand these to the doctor at the beginning of the consultation. Question sheets were given to all patients attending one clinic over a 13 week period. STRATEGIES FOR CHANGE: Use of sheets: number of patients taking or refusing a sheet, with reasons for refusal, were recorded. Doctors noted those who handed questions sheets to them Satisfaction with sheets: patients completed a short feedback form after the consultation Effect on consultations: evaluated through interviews with the doctors. EFFECTS OF CHANGE: In total, 66 (41%) of the 162 patients offered the sheet declined: 14 had already prepared questions; eight being seen for the first time felt they did not know what to ask. Seventeen had used the sheet on a previous visit and did not need it again; 19 gave no reason; the rest said they had no questions. Seventy six (47%) patients produced a sheet in the consultation. Of those using the sheet, 64 (84%) liked it and 61 (80%) found it useful. Fifty two (68%) wished to use it at future consultations. The doctors reported that through questions articulated on the sheets many issues, fears, and misunderstandings emerged which otherwise would not have been identified. Concerns about increasing consultation time and clinical disruption did not materialize. In contrast, doctors reported patients to be taking more initiative and control, particularly on subsequent visits. None of these changes was noted in the comparison clinic. LESSONS LEARNT: An attractive, clear question sheet proved a simple but effective intervention in the consultation. Parents felt empowered to take control. The approach may have wider applicability, but implementation requires staff training and support to ensure its continuing use; this ensures medical staff adjust to a new consultation format, and that clinic nurses see the value of the sheets and continue to provide them. PMID- 10848370 TI - Adverse events in health care: issues in measurement. PMID- 10848371 TI - Public disclosure of performance data: learning from the US experience. PMID- 10848372 TI - Disseminating healthcare information: getting the message across. PMID- 10848373 TI - Development of an audit instrument for nursing care plans in the patient record. AB - OBJECTIVES: To develop, validate, and test the reliability of an audit instrument that measures the extent to which patient records describe important aspects of nursing care. MATERIAL: Twenty records from each of three hospital wards were collected and audited. The auditors were registered nurses with a knowledge of nursing documentation in accordance with the VIPS model--a model designed to structure nursing documentation. (VIPS is an acronym formed from the Swedish words for wellbeing, integrity, prevention, and security.) METHODS: An audit instrument was developed by determining specific criteria to be met. The audit questions were aimed at revealing the content of the patient for nursing assessment, nursing diagnosis, planned interventions, and outcome. Each of the 60 records was reviewed by the three auditors independently and the reliability of the instrument was tested by calculating the inter-rater reliability coefficient. Content validity was tested by using an expert panel and calculating the content validity ratio. The criterion related validity was estimated by the correlation between the score of the Cat-ch-Ing instrument and the score of an earlier developed and used audit instrument. The results were then tested by using Pearson's correlation coefficient. RESULTS: The new audit instrument, named Cat ch-Ing, consists of 17 questions designed to judge the nursing documentation. Both quantity and quality variables are judged on a rating scale from zero to three, with a maximum score of 80. The inter-rater reliability coefficients were 0.98, 0.98, and 0.92, respectively for each group of 20 records, the content validity ratio ranged between 0.20 and 1.0 and the criterion related validity showed a significant correlation of r = 0.68 (p < 0.0001, 95% CI 0.57 to 0.76) between the two audit instruments. CONCLUSION: The Cat-ch-Ing instrument has proved to be a valid and reliable audit instrument for nursing records when the VIPS model is used as the basis of the documentation. PMID- 10848374 TI - Quality improvement around the world: how much we can learn from each other. AB - The USA National Forum on Quality Improvement in Health Care--organised by the Institute of Healthcare Improvement (Boston USA)--attracts many people from outside North America. At the 1999 meeting 20 countries were represented. A session on "Quality improvement around the world" was included in the pre conference programme to bring together people working in many countries to explore and compare their experiences in a programme of short presentations (table 1). This article draws together some of the themes that emerged from the presentations and from the discussion. PMID- 10848375 TI - Drug treatments for schizophrenia. PMID- 10848377 TI - Figuring out the fruit fly. A step closer to reading the human genome. PMID- 10848376 TI - A framework for effective management of change in clinical practice. PMID- 10848378 TI - Midair attacks. What airlines do when passengers need help with medical emergencies. PMID- 10848379 TI - How to get to your golden years. Health: boomers still have time to choose between decrepitude and a vigorous old age. But time is running out. PMID- 10848380 TI - Looking beyond Viagra. The 30 million American men with impotence problems may soon have two oral treatments to choose from. How do they compare? PMID- 10848381 TI - Does my child need Ritalin? Stimulants are still the most effective treatment for ADHD. The challenge is to use them wisely. PMID- 10848382 TI - The patients' perspective on clinical governance. PMID- 10848383 TI - Incomplete excision of basal cell carcinomas: an audit. AB - The Plastic Surgery Department undertook an audit of the rate of excision of basal cell carcinomas (BCCs) at St George's Hospital. The aim of the audit was to determine the rate of incomplete excision of BCCs excised by the department over a six-month period and compare it with national audit figures. The rate of incomplete excision was 13.73 per cent or seven out of 51 BCCs, which compared favourably with the rates quoted in the literature. No change in the management of basal cell carcinomas was made. PMID- 10848384 TI - Evaluation of an orthopaedic screening service in primary care. AB - Historically, provision of orthopaedic services has been hospital based with GPs referring patients for specialist opinion. Growing demands on the service have led to new initiatives to reduce waiting times. One such initiative has been the introduction of orthopaedic assistants, usually physiotherapists, working with an extended scope of practice who see patients after referral to secondary care and determine the patients' ongoing management. Studies to date have examined the effect of an orthopaedic assistant working alongside a consultant in the hospital environment. This study describes the impact on the management of the orthopaedic caseload in one general practice resulting from "screening" prior to referral to secondary care by a physiotherapist with an extended scope of practice. It demonstrates the successful management of the majority of patients within primary care. PMID- 10848385 TI - The relationship between research and service development: an illustrative example of a pilot study introducing complementary therapies into primary care. AB - In this paper the commissioners of an evaluation and the researchers jointly review the relationship between research and service development at a local level in an evaluated health authority pilot project to introduce complementary therapies into primary care. The article discusses the importance of organisational arrangements between the research and the service development, focusing on the close working relationship between researchers and stakeholders in the research and corresponding service development. The relationship between the research and service development was not linear and the benefits were not based solely on outcomes of the treatment but also on the ways the evaluation gave insight into how the pilot service was delivered. Factors such as personal commitment to the project and close working relationships by all concerned are important. These elements are rarely emphasised but have to be taken into account if evidence-based health care is to achieve its potential. PMID- 10848386 TI - Is day case tonsillectomy suitable for children in Cumbria? AB - Day case tonsillectomy has been advocated as a means of reducing health-care costs associated with in-patient care. The authors studied 74 consecutive children undergoing conventional overnight stay tonsillectomy using a parental questionnaire and a retrospective case note review. Medical exclusion criteria for day surgery were present in 21 per cent, and social exclusion criteria in 82 per cent. Overall, only 16 per cent of children satisfied all the criteria for suitability. On the day of surgery, 29 per cent of children were reported as having poorly controlled pain and 31 per cent as having nausea and vomiting, such that unplanned admission would have been likely had they been done as day cases. In total 63 per cent of parents were unhappy or very unhappy with the possibility of same day discharge. The authors do not plan to introduce day case tonsillectomy in Cumbria. PMID- 10848387 TI - The care pathway: a tool to enhance clinical governance. AB - A care pathway defines in detail the individual components of treatment for a group of patients. A well-written pathway can lead to consistent care of the highest quality. There are both educational and audit advantages to the approach. The authors detail the means by which groups are selected and the care pathways written. An example of the benefits that can accrue comes from their experience at Ashford Hospital of running a pathway for patients having prostate surgery. The pathways are appreciated by patients, nurses, doctors and managers. Care is improved, costs contained and clinical governance enhanced. PMID- 10848388 TI - The Internet: tomorrow's portal to non-traditional health care services. AB - It is not at all surprising that people are not enamored of hospitals, as they are almost always associated with negatives such as pain, loss of mobility, and high cost. On a less emotional level, hospitals are perceived as big businesses, and consumers are getting inured to a consistently decreasing level of caring and service, particularly from "service" businesses. Many people are going to the Internet for health information due to the belief that today's doctor-patient relationship lacks an attention to detail and personal touch that was there in the past. The Internet is empowering consumers. If it continues this way, consumers will rely on physicians less and shop for alternatives more. In this article, the author makes the following points: (1) Consumer attitudes about traditional health care providers are changing significantly as their desire for more involvement in and control over the management of their health increases. This desire increases their need for information. (2) The Internet is proving to be the information source consumers need. Anyone with a modem-equipped personal computer and a telephone line can access health care and scientific information on Web sites ranging from Medline and the Merck Manual to Ask Dr. Dean and Dr. Ruth. (3) There are alternatives to traditional providers and methods, and consumers can not only find them on the Internet, but also get information that is stated objectively and non-technically. PMID- 10848389 TI - The medical records professionals' view of the electronic medical record. AB - The migration of Kaiser-Permanente (KP), a large managed care organization, to the management of health care information in a paperless environment is presented from the viewpoint of Health Information Management (HIM) professionals. The author conducted an informal survey of these professionals and presents a synopsis of their recommendations in the context of KP's efforts to develop an enterprise Electronic Medical Record. Their consensus is that physicians are primarily information managers and that successful health care organizations will be those who most effectively manage their information. PMID- 10848390 TI - Toward an integrated computerized patient record. AB - Developing a comprehensive electronic medical record system to serve ambulatory care providers in a large health care enterprise requires significant time and resources. One approach to achieving this system is to devise a series of short term, workable solutions until a complete system is designed and implemented. The initial solution introduced a basic (mini) medical record system that provided an automated problem/summary sheet and decentralization of ambulatory-based medical records. The next step was to partner with an information system vendor committed to continued development of the long-term system capable of supporting the health care organization well into the future. PMID- 10848391 TI - Computerized medical record war story: practical experiences of computer implementation at the front. AB - Computerized medical records will be the norm for all medical practices and soon. This article is an actual account of the successful implementation of a computerized medical records documenting and coding solution at a cardiology practice. Computerizing a medical office is a continuing and evolving process that involves setting-up, training and rethinking. The computer network IS the patient chart. The physicians and staff must be in the information loop and use the information tool, the computer, to record and retrieve medical information immediately and effectively. Medicine is just now on the threshold of realizing the immense benefit that the computer will bring to it. PMID- 10848392 TI - Making EMRs really work: the Southeast Health Center experience. AB - While electronic medical records (EMRs) have long held the promise of improved efficiency and quality, they have not yet made significant inroads into ambulatory care. One reason may be attempted EMR implementation in "old paradigm" clinic models. Southeast Health Center (SEHC) in Indianapolis, Indiana, began by reengineering its clinic procedures; only then did staff look for a computerized record. Because the new clinic model meshed nicely with the design of a relatively new EMR, a joint project was undertaken to test this EMR in SEHC's unique environment. This article details the reengineering, EMR implementation, project dynamics, results, and lessons learned. PMID- 10848393 TI - Computer-based ambulatory information systems: recent developments. AB - This article updates the author's earlier review of some of the major computer based ambulatory information systems and the literature evaluating their costs, benefits, effect on quality of care, and physician acceptance. The evidence suggests that computer-based information systems can increase access to clinical information, improve physician performance, enhance quality of care, and facilitate outcomes research review. In addition to presenting health information networks and clinical decision support systems such as reminder systems, drug ordering systems, and medical care management systems, the article describes applications of telemedicine and Web-based systems. It also discusses barriers to the widespread use of computer-based ambulatory information systems. PMID- 10848394 TI - Is the physician office the wild, wild west of health care? AB - Elective, office-based surgery has captured the interest of consumers and, more recently, the attention of state health care regulatory agencies. In most states today, patients can undergo cosmetic surgery, liposuction, endoscopy, colonoscopy, microlaparoscopy, and various other procedures requiring sedation or anesthesia in physician offices even though no regulatory safeguards that would ordinarily benefit patients in accredited or licensed facilities exist. Media accounts of deaths and serious injuries associated with liposuction and anesthesia performed in physician offices resulted in legislative and regulatory initiates, such as those in California and New Jersey. Increased regulatory oversight, changes in patterns of reimbursement, and greater consumer awareness of safety and quality-of-care issues should aid in reducing the risks of office based surgery. PMID- 10848395 TI - Walking wounded: the status of ambulatory care in East Timor. PMID- 10848396 TI - Digital health care--the convergence of health care and the Internet. AB - The author believes that interactive media (the Internet and the World Wide Web) and associated applications used to access those media (portals, browsers, specialized Web-based applications) will result in a substantial, positive, and measurable impact on medical care faster than any previous information technology or communications tool. Acknowledging the dynamic environment, the author classifies "pure" digital health care companies into three business service areas: content, connectivity, and commerce. Companies offering these services are attempting to tap into a host of different markets within the health care industry including providers, payers, pharmaceutical and medical products companies, employers, distributors, and consumers. As the fastest growing medium in history, and given the unique nature of health care information and the tremendous demand for content among industry professionals and consumers, the Internet offers a more robust and targeted direct marketing opportunity than traditional media. From the medical consumer's standpoint (i.e., the patient) the author sees the Internet as performing five critical functions: (1) Disseminate information, (2) Aid informed decision making, (3) Promote health, (4) Provide a means for information exchange and support--the community concept, and (5) Increase self-care and manage demand for health services, lowering direct medical costs. The author firmly submits the Web will provide overall benefits to the health care economy as health information consumers manage their own health problems that might not directly benefit from an encounter with a health professional. Marrying the Internet to other interactive technologies, including voice recognition systems and telephone-based triage lines among others, holds the promise of reducing unnecessary medical services. PMID- 10848397 TI - The bottom-line case for making house calls. PMID- 10848398 TI - Being Dr. Nice Guy can backfire. PMID- 10848399 TI - Let's stop giving alternative medicine a free ride. Interview by Mark Crane. PMID- 10848400 TI - Welcoming the menopausal woman to your practice. PMID- 10848401 TI - Your duty of care when on call to the ER. PMID- 10848402 TI - Do doctors give HMO patients a fair shake? PMID- 10848403 TI - Treating pain: can doctors put their fears to rest? PMID- 10848404 TI - Pick the team, and write the game plan. PMID- 10848405 TI - My patient's car is his castle. PMID- 10848406 TI - Folk medicine deserves our respect. PMID- 10848407 TI - Health spending is soaring. What's so bad about that? PMID- 10848408 TI - The legal perils in caring for the elderly. PMID- 10848409 TI - Doctors and the Web. Help your patients surf the Net safely. AB - The Internet promises to touch every aspect of a physician's professional life, from patient relations to access to clinical studies, from billing to patient records, from marketing to e-mail. To help you make sense of what may be the most profound force in medical practice today, we're kicking off a new series with this article on helping patients navigate the Internet. Future installments, which will run in our first issue of every month, will look at such topics as online patient charts; Web-based electronic medical records; services that electronically connect doctors with health plans, hospitals, and other providers; and online supply purchasing. PMID- 10848410 TI - Could the doctors heal our baby's heart? PMID- 10848411 TI - How the animal rights zealots threaten medical progress. PMID- 10848412 TI - Malpractice: who's on the hook when a hospitalist discharges your patients? PMID- 10848413 TI - Managed care holdouts: how are they doing now? PMID- 10848414 TI - HMNo: the name of this practice says it all. PMID- 10848415 TI - Patients helped me keep my promise to the poor. PMID- 10848416 TI - Taking a picture of a mind gone awhirl. New imaging method bolsters ADHD diagnosis. PMID- 10848417 TI - Of genes, grain, and grocers. The risks and realities of engineered crops. PMID- 10848418 TI - No blackout required. After years of decline, the number of births goes up. PMID- 10848419 TI - Your medical school may be suffering from ill health. PMID- 10848420 TI - What should CT's role be in pediatric appendicitis? PMID- 10848421 TI - A look at skipped findings that turn out to be cancer. PMID- 10848422 TI - Low-dose CT screening reduces lung cancer deaths. PMID- 10848423 TI - International studies make case for high mammo volume. PMID- 10848424 TI - Don't forget about training in optimizing value of new technology. PMID- 10848425 TI - New indications give PET a lift after struggle over regulatory and reimbursement issues. PMID- 10848426 TI - MRI a standout for evaluation of abused knee joints. PMID- 10848427 TI - MR and PET assist in workup of patients with epilepsy. PMID- 10848428 TI - The Internet-linked laboratory: fundamentally changing the delivery of laboratory information and results. PMID- 10848429 TI - The pathologist: prime candidate for principal investigator in the deconstruction of disease. PMID- 10848430 TI - Graduate education in clinical systems: distance learning in an emerging field. PMID- 10848431 TI - An automated, rapid ELISA for the determination of D-dimer in the evaluation of venous thrombosis. PMID- 10848432 TI - Performance evaluation of a hematology blood counter with five-part leukocyte differential capability. AB - The summarized data and other studies to date indicate that performance of the Pentra 60 is comparable to other hematology analyzers for all CBC and 5-Diff parameters. Comparative studies indicate good correlation for all the reportable CBC parameters, lymphocyte counts, and neutrophil counts with 24 hr stability on blood samples. Good intermethod correlations on monocyte and eosinophil counts were observed. Only basophil counts showed poor intermethod correlations, but this is expected on a statistical basis, and the counts are similar to those reported for other hematology analyzer performance studies. The combination of the MDSS and stepper motor fluidic system allows for low-volume blood sampling, compact size, and low operational noise level. The Pentra 60 is well suited for physician's office laboratories, medical clinics, small- or medium-size hospitals with less than 100 CBCs per day, and larger hospitals or reference laboratories that need back-up for a high-end automated hematology analyzer. PMID- 10848433 TI - Using automated prenatal screening assays in the clinical laboratory. PMID- 10848434 TI - Evaluating test performance criteria: concepts and practices. Part 4: The assignment of performance limits and the probability of total error compliance. PMID- 10848435 TI - A point-of-care analyzer for blood gases, Chem 7, Hct, and Hgb. AB - A single POC analyzer can satisfy the entire test menu needs of clinicians in a wide variety of settings, including OR, ED, ICU, and NICU. A single POC platform provides complete diagnostic information, reduces turnaround time to less than 2 min, reduces blood loss, and streamlines the performance of POC testing. For the laboratory, a single platform simplifies implementation and management of POC testing while increasing the tasks of meeting regulatory compliance. PMID- 10848437 TI - A rapid, small, semiautomated nephelometer. PMID- 10848436 TI - Dynamic management opportunities in diagnostic/clinical services. PMID- 10848438 TI - Recent developments in humanized MAbs for immunotherapy: a personal tribute to Dr. Martin Lockwood. PMID- 10848439 TI - Increased anticoagulant osmolality improves separation of leukocytes from red blood cells (RBC). AB - BACKGROUND: The bottom-and-top (BAT) procedure separates the buffy coat (BC) from plasma and red blood cells (RBC). The contents of mononuclear cells (MNC) remaining in the RBC are about 1 x 10(6) cells/unit, whereas the granulocytes are removed less effectively, 500-800 x 10(6) or more remaining in the RBC unit. The aim was to improve the separation efficacy by collecting the blood in an hyperosmolar anticoagulant, followed by BAT separation. It was expected that the red cells would shrink, thereby increasing their density, while the granulocytes would not change volume and density. STUDY DESIGN AND METHODS: 18 donors were included in the study, 12 in the test group and 6 in the control group. CPD-SAGM bags were used, with a modification of the anticoagulant by removal of 20-ml CPD from all units and addition of 20-ml mannitol (test group) or 20 ml of isotonic saline (control group). The collected blood units were cooled on butanediol plates for 2-4 h, then centrifuged and separated into components. The levels of leukocytes in the whole blood, the BC and the RBC were determined by flow cytometry gated for intact CD45+ cells. A number of other tests were performed during 42-day storage. RESULTS: The plasma yield was slightly higher in the test group than in the control group (ns). The contents of leukocytes (CD 45+ intact cells) in the RBC units were 32 +/- 20 x 10(6) in the test group and 573 +/- 241 x 10(6) in the control group. The numbers of MNC were 1.2 +/- 0.6 x 10(6) and 2.6 +/- 1.8 x 10(6), respectively. The RBC 2,3-DPG concentration was slightly better maintained in the test group at day 7 of refrigerated storage (p = 0.0027), but most other tested parameters showed no difference during 42-day storage. It was possible to prepare platelet concentrates with good yield using the pooled-BC method. CONCLUSION: This study indicates that considerable improvement in the BAT procedure can be obtained if the anticoagulant is made hypertonic by the addition of mannitol. This is achieved without altering the already low levels of MNC and keeping the same quality. PMID- 10848440 TI - Cascade filtration for TTP: an effective alternative to plasma exchange with cryodepleted plasma. AB - TTP remains enigmatic both in terms of etiology and management. The most recent approach is aggressive plasma exchange (PE) employing cryopoor plasma for replacement, based on the pathogenetic relevance given to exceedingly large Von Willebrand (VWF) multimers in the determination of the syndrome with normalization during remission. PE with fresh frozen plasma (FFP) is better than FFP infusion as shown by a recent Canadian study, supporting the theory that to treat TTP an offending circulating agent needs to be removed from the patient's plasma in contrast to the hypothesis that a missing factor is to be given along with FFP. A more recent hypothesis is supported by the results of studies published by the end of 1998 [Moake J, Chintagumpala M, Turner N et al. Blood 1994;84:490-97; Moake J, McPherson PD. Am J Med 1989;87: 3-9N] which would show that TTP is mediated by auto-antibodies to VWF-cleaving protease, or is the result of deficiency of the protease ascribed to abnormalities in its production, function or survival. Plasmapheresis without plasma infusion is relatively ineffective perhaps because it does not increase the protease activity. Cascade filtration (CF) is the autologous counterpart of plasmapheresis. It has been used by our group since 1980 to remove from patients plasma macromolecules such as VWF, fibrinogen, LDL and circulatory immune complexes (CIC). After secondary filtration, the autologous plasma has a composition which is very similar to that of allogeneic plasma after cryoprecipitation, a product which used in the management of TTP. Based on this knowledge, in 1994 we began to use CF in the treatment of TTP patients. In the beginning (7 patients) CF was combined with a decreasing number of conventional PEs using allogeneic plasma for substitution. Lately only CF with some plasma supplementation has been used in the last 9 cases. From a clinical point of view our 16 patients achieved remission after a number of treatments (11 +/- 7) that compares sufficiently well with those required by our historical control group of 47 cases (14 +/- 13). Of course the patient's exposure to allogenic plasma was significantly lower for patients in the CF only group (1.4 +/- 1.2 plasma U/session) compared to the PE + CF group (4.4 +/- 2.3 plasma U/session) or for the controls treated by PE only (10.8 +/- 4.6 plasma U/session). There were no deaths in the CF or PE + CF groups and no untoward effect was observed. On the contrary there were 5 deaths (1 on the day of presentation) in the PE group, and 1 HBV and 2 HCV infections as well as 4 severe allergic reactions to plasma proteins (or passive antibody infusion). We conclude that CF is presently the best treatment to offer to patients suffering from sporadic TTP and that CF may contribute to expanding the knowledge of the pathogenetic mechanisms of this uncommon multisystem disorder. PMID- 10848441 TI - Improvement of visual acuity in patients suffering from diabetic retinopathy after membrane differential filtration: a pilot study. AB - BACKGROUND: Membrane differential filtration is an extracorporeal treatment procedure which eliminates high molecular weight proteins and lipids from the blood. This pilot study was initiated to investigate the short-term effects on blood rheology and visual function in patients suffering from diabetic retinopathy. METHODS: Six patients with non-proliferative diabetic retinopathy, clinically significant macular edema (five patients, nine eyes) and inactive proliferative diabetic retinopathy (one patient, one eye) underwent a single treatment with a hollow fiber secondary plasma filter. The main parameter measured was visual acuity prior to and after treatment. Biochemical and rheological parameters (whole blood and plasma viscosity, and erythrocyte aggregation) were also measured. The mean follow-up was 25 (range 4-90) days after treatment. RESULTS: The mean improvement of visual acuity was 1.4 lines (SD 0.8 lines, p = 0.002) 24 h after therapy. This remained stable during the follow up period. The rheological measures were significantly lowered. A significant reduction of total protein, fibrinogen, IgG, IgM, IgA, alpha-2-macroglobulin, total cholesterol, LDL and HDL was found. CONCLUSION: This study demonstrates the rheological impact of membrane differential filtration. It was shown that rheological changes correlated with clinical improvement in patients suffering from diabetic retinopathy with clinically significant macular edema. PMID- 10848442 TI - The correlation between erythropoiesis and thrombopoiesis as an index for pre operative autologous blood donation. AB - We have previously reported that the platelet count and an increase in platelet number reflect individual erythropoietic capacity in pre-operative autologous blood donation (PABD). We have examined the correlation between erythropoiesis and thrombopoiesis by quantitative in vitro determination of thrombopoietin (TPO), erythropoietin (EPO), and interleukin-6 (IL-6) in patients with PABD. A sequential increase in platelet count with donation could not be explained by an increase in TPO. TPO showed a tendency to be inversely related to the pre donation platelet count, and to be related to the pre-donation hemoglobin level. There was an inverse relationship between the TPO and EPO levels. As seen with these results, a high erythropoietic state induces restraint of thrombopoiesis, and a low erythropoietic state induces an increase in thrombopoiesis. These effects modulate EPO and TPO via negative feedback. These results provide some practical important information for performing autologous blood donation. Further studies are needed to elucidate the details of these correlations. PMID- 10848443 TI - What's happening? Current status of high dose chemotherapy in breast cancer. AB - To date, no definite answers are available to the initial question, as to whether or not HDCT and ASCT can improve the quantity or quality of life. Completing the available prospective, randomized phase III studies; using HDCT earlier in the course of treatment, applying drugs that are active against breast cancer, comparing the experimental treatment to standard therapy, and using appropriate sample size to detect clinically meaningful and statistically significant differences are all paramount requirements to answer the question successfully. Within the next few years, a number of European [13] and two Canadian studies [6,14] that are currently accruing patients, will contribute important information regarding the role of HDCT and ABMT for breast cancer. PMID- 10848444 TI - What's happening? Protein A columns: current concepts and recent advances. PMID- 10848445 TI - 12th Congress of the European Society of Apheresis. Abstracts. PMID- 10848446 TI - Setting sail on the US Chronic Care. PMID- 10848447 TI - Maximizing quality of life. PMID- 10848448 TI - Thinking outside the box. PMID- 10848449 TI - Promoting the healing process. PMID- 10848450 TI - Using a team approach. PMID- 10848451 TI - Widening the scope. PMID- 10848452 TI - Practice guidelines. PMID- 10848453 TI - Maintaining mobility. PMID- 10848454 TI - Adaptable living. PMID- 10848455 TI - Filling a need. PMID- 10848456 TI - For women only. PMID- 10848457 TI - Evaluating WIC. AB - The Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) has enjoyed extraordinary bipartisan support stemming from the widespread belief that research studies have proven that WIC "works." Although some studies suggest real dietary and health improvements, the greatest benefits only apply to WIC's prenatal program (just a small part of the total program). Even here, weaknesses in the research render the findings highly uncertain. The three most significant weaknesses are (1) selection bias, (2) simultaneity bias, and (3) lack of generalizeability. The resulting uncertainty places WIC's possible impacts on infant mortality, prematurity, and birthweight on a range from zero to substantial. For infants, children, and postpartum and breastfeeding mothers, the only impacts seem to be small to modest effects on anemia and nutrient intake. This paper does not argue that WIC's weaknesses justify abandoning or even cutting the program. On the contrary, there should be a sustained effort to make the program more effective. This effort should start with a policy debate about WIC's role and impacts, coupled with a grant of greater flexibility to state and local WIC agencies to open the program to innovation and experimentation. To increase WIC's positive impacts, we propose a series of possible reforms, each to be thoroughly evaluated. PMID- 10848458 TI - Correlation bias in U.S. census adjustment. AB - One form of error that can affect census adjustments is correlation bias, reflecting people who are doubly missing--from the census and from the adjusted counts as well. This article presents a method for estimating the total national number of doubly-missing people and their distribution by race and sex. Application to the 1990 U.S. census adjustment leads to an estimate of 3 million doubly-missing people. Correlation bias is likely to be a serious problem for census adjustment in 2000. The methods of this article are well suited for measuring its magnitude. PMID- 10848459 TI - A randomized field trial in Oakland, California. AB - This study evaluates the impact of a civil remedy program (Beat Health) on drug and disorder problems under experimental field trial conditions. Using calls for service data, the authors find statistically significant differences between the control and experimental groups, when drug problem calls prior to the start of the intervention were compared to drug calls during a 12-month follow-up period. The Beat Health sites, especially the residential sites, improved relative to the control sites. By contrast, the control treatment (patrol response) led to significant increases in drug problems, particularly at the commercial properties included in this study. The authors also examined the spatial influences of the Beat Health and patrol responses in catchment areas surrounding each of the 100 study sites. The results show some improvement in the experimental residential sites. The authors found, however, a possible displacement of drug problems in and around both the commercial experimental and control sites. They show that the displacement effect is most notable in the commercial control sites. PMID- 10848460 TI - A case of provider angst. PMID- 10848461 TI - Making it in the new millennium. PMID- 10848462 TI - Ground rules for online services. PMID- 10848463 TI - Unlocking the secrets of the alcoholic brain. PMID- 10848464 TI - Treatment planning goes national. PMID- 10848465 TI - The power of the TC. PMID- 10848467 TI - Charter's collapse leaves field short. PMID- 10848466 TI - Accountability in the public AOD system. National Association of State and Drug Abuse Directors. PMID- 10848468 TI - What would it take to get you to confess? PMID- 10848469 TI - The comeback plague. Once nearly conquered, TB is rebounding in menacing new strains. PMID- 10848470 TI - Five little piggies going to market. Will clones be a source of organs for humans? PMID- 10848471 TI - Some kids' bizarre compulsions may get their start from a common germ. PMID- 10848472 TI - Before you get lasered, squint at the fine print. PMID- 10848473 TI - Drug trials are clamoring for kids, but scrutinize the study before signing up. PMID- 10848474 TI - Smoking is the leading preventable cause of death and disability in the United States. PMID- 10848475 TI - Effective use of mass media to prevent cigarette smoking. PMID- 10848476 TI - Florida's "truth" campaign: a counter-marketing, anti-tobacco media campaign. AB - The "truth" campaign was created to change youth attitudes about tobacco and to reduce teen tobacco use throughout Florida by using youth-driven advertising, public relations, and advocacy. Results of the campaign include a 92 percent brand awareness rate among teens, a 15 percent rise in teens who agree with key attitudinal statements about smoking, a 19.4 percent decline in smoking among middle school students, and a 8.0 percent decline among high school students. States committed to results-oriented youth anti-tobacco campaigns should look to Florida's "truth" campaign as a model that effectively places youth at the helm of anti-tobacco efforts. PMID- 10848477 TI - Expert opinions on optimal enforcement of minimum purchase age laws for tobacco. AB - A questionnaire on how youth access laws should be enforced was sent to 20 experts who had administered and/or evaluated a youth access enforcement program. Respondents agreed on the need for a high level of retail compliance, checkers representative of the community, checks at least twice per year, a graduated penalty structure with license revocation, and bans on self-service and vending machines. Respondents indicated the need for research on the effects of ID use, frequency of checks, penalty structures, and the effects on smoking rates of youth access policies alone and in conjunction with other tobacco control policies. PMID- 10848478 TI - The impact of untruthful age reporting during tobacco compliance checks. AB - A pilot study consisting of 76 compliance checks was conducted in October 1998 in Chautauqua County, New York, by a 15-year-old white female. Half of the merchants were randomized to be checked under a standard protocol whereby the minor was instructed to answer truthfully when asked about her age. The remaining stores were checked under a new protocol that permitted the minor to state that she was 18 years old if asked about her age. It was found that untruthful age reporting during compliance checks has little impact on the overall compliance rate. PMID- 10848479 TI - Planning and initiation of the ONDCP National Youth Anti-Drug Media Campaign. AB - An alarming upsurge in substance use among American youth in the past decade prompted the Office of National Drug Control Policy (ONDCP) to organize the multifaceted, primary prevention National Youth Anti-Drug Media Campaign. The campaign focuses on adolescents (especially ages 11-13 years), but also targets adolescents' parents and other influential adults. The campaign's main goal is to educate and enable American youth to reject illegal drugs. The purpose of this article is to describe the origin of campaign objectives, processes for the development of specific advertising messages to fulfill these objectives, and how the ONDCP experience could provide a model for other health-oriented media campaigns. PMID- 10848480 TI - A content analysis of television advertising for the Massachusetts Tobacco Control Program media campaign, 1993-1996. AB - Television advertising for the Massachusetts Tobacco Control Program (MTCP) was launched in 1993. The campaign's first three years were examined against an analytical framework based in behavioral change and communications theory. The most common focus in the 49 advertisements was tobacco industry practices, followed by health consequences for smokers and smoking prevention. Only one smoking prevention advertisement was targeted to children younger that age 13. Only four advertisements mentioned policy actions to reduce youth access or curtail exposure to environmental tobacco smoke. Only two advertisements listed the telephone number for the MTCP's smoking cessation program. The television campaign was uncoordinated with the MTCP's community-level programs. PMID- 10848481 TI - Adult and youth response to the Massachusetts anti-tobacco television campaign. AB - This article is a response to DeJong and Hoffman's critique of the Massachusetts anti-tobacco television advertisements. It presents data on the recall and perceived effectiveness of the advertisements by a representative sample of adults and youth, and summarizes a previously published analysis of the impact of exposure to the advertisements on progression to regular smoking among youth. These data indicate that the campaign has achieved high levels of penetration into the population, that the advertisements are seen by the public to be effective, and that high levels of reported exposure are associated with reductions in teen smoking. PMID- 10848482 TI - Development of a low-budget tobacco prevention media campaign. AB - Utah joined the ranks of other states that have passed a tobacco tax to fund a tobacco control program, including a media campaign. However, unlike other states working with multimillion dollar campaigns, $250,000 was allocated toward an anti tobacco media campaign. Formative research guided the strategic development of the media campaign. PMID- 10848483 TI - The industry manipulation attitudes of smokers and nonsmokers. AB - A cross-sectional random sample was obtained of Florida youth between the ages of 12-17. Data were collected through a telephone survey after obtaining parent and child consent. Industry manipulation attitudes of three groups (self-identified nonsmokers who did not use cigarettes in the past 30 days, self-identified nonsmokers who used cigarettes in the past 30 days, and self-identified smokers who used cigarettes in the past 30 days) were compared. Constraints resulting from the method of data collection resulted in a conservative estimate of the strength of the association between industry manipulation attitudes and smoking behavior. PMID- 10848484 TI - Developing an ATOD prevention campaign for Asian and Pacific Islanders: some considerations. AB - This article provides an overview of the Asian and Pacific Islander (API) populations in the United States as it relates to developing targeted alcohol, tobacco, and other drug (ATOD) prevention media. Although APIs represent several ethnic subgroups, many API communities face the same risk factors including pressure on youth to achieve, immigration and acculturation stressors, shame and denial in addressing substance abuse, and racism and discrimination. The authors recommend that media prevention campaigns targeting API communities emphasize cultural and family strengths and the active involvement of parents. The article concludes with an introduction to eight API communities in the United States. PMID- 10848485 TI - Background and principles of an African American targeted national anti-drug campaign. AB - This article provides a review of key African American community values drawn from the literature. Central to this article is how these values can be incorporated into anti-drug messages and materials targeted to African Americans as part of a national five-year campaign being implemented by the White House Office of National Drug Control Policy and the Partnership for a Drug-Free America. The campaign is the first national campaign to base messages on behavioral science research and theories and subsequently uses research about targeted audiences to design prevention messages that are culturally relevant. Specific implications for the format and content of messages directed to African Americans are discussed. PMID- 10848486 TI - Strategies for an effective youth counter-marketing program: recommendations from commercial marketing experts. AB - Intensive and sustained efforts to "counter-market" tobacco among teenagers are necessary to negate the "friendly familiarity" created by tobacco advertising and to communicate the true health and social costs of tobacco use. Counter-marketing campaigns should: highlight a tobacco-free lifestyle as the majority lifestyle of diverse and interesting individuals; explain the dangers of tobacco in a personal, emotional way; offer youth empowerment and control; use multiple voices, strategies, and executions; offer constructive alternatives to tobacco use; and portray smoking as unacceptable and undesirable for everyone. Counter marketing activities should work in concert with other interventions to alter social norms regarding tobacco. PMID- 10848487 TI - Developing a theory-based anti-drug communication campaign for Hispanic children and parents. AB - Recent research suggests some alarming substance abuse trends among Hispanic/Latino adolescents. Children in this age group are at a particularly vulnerable juncture in their lives, and early prevention efforts are warranted. For a national anti-drug campaign targeting this young Hispanic/Latino audience and their parents, the White House Office of National Drug Control Policy and the Partnership for a Drug-Free America are incorporating culturally appropriate messages based on behavioral science research and theories. The purpose of this article is to provide information and recommendations on relevant factors and influences in the Hispanic/Latino culture that should be addressed in designing and implementing this campaign. PMID- 10848488 TI - Planning an effective anti-smoking mass media campaign targeting adolescents. AB - This article addresses the following issues: Can an anti-smoking campaign that depends largely on mass media vehicles effectively reduce adolescent tobacco use? Why is an integrated campaign recommended and what are the steps in designing such a campaign? How should the campaign be evaluated? Specific topics include recommended campaign expenditures, target audience identification, selection of persuasive message content, executional (stylistic) considerations, media buying decisions, the use of focus group research and advertising copy-testing research, and outcome evaluations. It is concluded that comprehensive strategic planning and extensive research at all phases of the campaign are essential to success. PMID- 10848489 TI - Gauging the effects of mass media policies: what do we need to know? AB - This article explores the components of an effective mass media policy and the role of mass media campaigns as part of a comprehensive set of tobacco control strategies. First, the main findings of the empirical literature are briefly reviewed. Then a framework for understanding mass media policies is developed showing the influence of concurrent tobacco control policies and the interrelationship between the two. A model will be presented that draws on concepts and research from the advertising and marketing literature, as well as from public health studies. A discussion of limitations of the extant literature and issues that warrant further examination will conclude. PMID- 10848490 TI - Axonal Charcot-Marie-Tooth disease and the neurofilament light gene (NF-L) PMID- 10848491 TI - Premature ovarian failure among fragile X premutation carriers: parent-of-origin effect? PMID- 10848493 TI - Data mining applied to linkage disequilibrium mapping. AB - We introduce a new method for linkage disequilibrium mapping: haplotype pattern mining (HPM). The method, inspired by data mining methods, is based on discovery of recurrent patterns. We define a class of useful haplotype patterns in genetic case-control data and use the algorithm for finding disease-associated haplotypes. The haplotypes are ordered by their strength of association with the phenotype, and all haplotypes exceeding a given threshold level are used for prediction of disease susceptibility-gene location. The method is model-free, in the sense that it does not require (and is unable to utilize) any assumptions about the inheritance model of the disease. The statistical model is nonparametric. The haplotypes are allowed to contain gaps, which improves the method's robustness to mutations and to missing and erroneous data. Experimental studies with simulated microsatellite and SNP data show that the method has good localization power in data sets with large degrees of phenocopies and with lots of missing and erroneous data. The power of HPM is roughly identical for marker maps at a density of 3 single-nucleotide polymorphisms/cM or 1 microsatellite/cM. The capacity to handle high proportions of phenocopies makes the method promising for complex disease mapping. An example of correct disease susceptibility-gene localization with HPM is given with real marker data from families from the United Kingdom affected by type 1 diabetes. The method is extendable to include environmental covariates or phenotype measurements or to find several genes simultaneously. PMID- 10848492 TI - Two genetic loci regulate T cell-dependent islet inflammation and drive autoimmune diabetes pathogenesis. AB - Insulin-dependent diabetes mellitus (IDDM) is a polygenic disease caused by progressive autoimmune infiltration (insulitis) of the pancreatic islets of Langerhan, culminating in the destruction of insulin-producing beta cells. Genome scans of families with diabetes suggest that multiple loci make incremental contributions to disease susceptibility. However, only the IDDM1 locus is well characterized, at a molecular and functional level, as alleleic variants of the major histocompatibility complex (MHC) class II HLA-DQB1, DRB1, and DPB1 genes that mediate antigen presentation to T cells. In the nonobese diabetic (NOD) mouse model, the Idd1 locus was shown to be the orthologous MHC gene I-Ab. Inheritance of susceptibility alleles at IDDM1/Idd1 is insufficient for disease development in humans and NOD mice. However, the identities and functions of the remaining diabetes loci (Idd2-Idd19 in NOD mice) are largely undefined. A crucial limitation in previous genetic linkage studies of this disease has been reliance on a single complex phenotype-diabetes that displays low penetrance and is of limited utility for high-resolution genetic mapping. Using the NOD model, we have identified an early step in diabetes pathogenesis that behaves as a highly penetrant trait. We report that NOD-derived alleles at both the Idd5 and Idd13 loci regulate a T lymphocyte-dependent progression from a benign to a destructive stage of insulitis. Human chromosomal regions orthologous to the Idd5 and -13 intervals are also linked to diabetes risk, suggesting that conserved genes encoded at these loci are central regulators of disease pathogenesis. These data are the first to reveal a role for individual non-MHC Idd loci in a specific, critical step in diabetes pathogenesis-T cell recruitment to islet lesions driving destructive inflammation. Importantly, identification of intermediate phenotypes in complex disease pathogenesis provides the tools required to progress toward gene identification at these loci. PMID- 10848495 TI - No evidence for parent of origin influencing premature ovarian failure in fragile X premutation carriers. PMID- 10848494 TI - Mapping of a new locus for autosomal recessive demyelinating Charcot-Marie-Tooth disease to 19q13.1-13.3 in a large consanguineous Lebanese family: exclusion of MAG as a candidate gene. AB - Autosomal recessive Charcot-Marie-Tooth disease (CMT) type 4 (CMT4) is a complex group of demyelinating hereditary motor and sensory neuropathies presenting genetic heterogeneity. Five different subtypes that correspond to six different chromosomal locations have been described. We hereby report a large inbred Lebanese family affected with autosomal recessive CMT4, in whom we have excluded linkage to the already-known loci. The results of a genomewide search demonstrated linkage to a locus on chromosome 19q13.1-13.3, over an 8.5-cM interval between markers D19S220 and D19S412. A maximum pairwise LOD score of 5.37 for marker D19S420, at recombination fraction [theta].00, and a multipoint LOD score of 10.3 for marker D19S881, at straight theta = .00, strongly supported linkage to this locus. Clinical features and the results of histopathologic studies confirm that the disease affecting this family constitutes a previously unknown demyelinating autosomal recessive CMT subtype known as "CMT4F." The myelin-associated glycoprotein (MAG) gene, located on 19q13.1 and specifically expressed in the CNS and the peripheral nervous system, was ruled out as being the gene responsible for this form of CMT. PMID- 10848496 TI - Premature ovarian failure is associated with maternally and paternally inherited premutation in Brazilian families with fragile X. PMID- 10848498 TI - Inflated false-positive rates in Hardy-Weinberg and linkage-equilibrium tests are due to sampling on the basis of rare familial phenotypes in finite populations. PMID- 10848499 TI - QTL fine mapping, in extreme samples of finite populations, for complex traits with familial correlation due to polygenes. PMID- 10848501 TI - Interpretation of linkage data for a Huntington-like disorder mapping to 4p15.3. PMID- 10848503 TI - Mechanisms regulating hypoxic respiratory depression during fetal and postnatal life. AB - Selected topics in the respiratory response to acute hypoxia in the fetus and newborn are reviewed. Peripheral chemoreceptors acting through ionotrophic glutamate receptors play an important role in affecting the initial augmentation phase. Whether fall off in peripheral chemoreceptor activity contributes to the secondary depressive phase remains controversial. A number of approaches including permanent electrolytic and reversible cooling lesions, Fos protein activation, and double-labeling immunohistochemistry has converged to show that an area in and around the locus ceruleus in the rostral pons affects the central depression. There is evidence that this is mediated by catecholamines acting at alpha(2)-adrenergic receptors. Tonic activity in early expiratory (postinspiratory) neurons may contribute to hypoxia-induced apneic episodes in the fetus and newborn. Desensitization of alpha-amino-3-hydroxy-5-methylisoxazole 4-proprionic acid receptors has been demonstrated in respiratory-related neurons both in vivo and in vitro. The role that this process might play in the depressive phase of the hypoxic ventilatory response has not been established. In vitro experiments with isolated brain stem-spinal cord preparations or transverse brain stem slices usually involve anoxia, whereas whole animal experiments use 8 15% O(2). Therefore, caution must be exercised in attempting to construct a unifying framework from these two approaches. PMID- 10848504 TI - Leptin, but not immune function, is linked to reproductive responsiveness to photoperiod. AB - Energetic demands are high while energy availability is minimum during winter. To cope with this energetic bottleneck, animals exhibit numerous energy-conserving adaptations during winter, including changes in immune and reproductive functions. A majority of individual rodents within a population inhibits reproductive function (responders) as winter approaches. A substantial proportion of small rodents within a species, however, fails to inhibit reproduction (nonresponders) during winter in the field or in the laboratory when maintained in winter-simulated day lengths. In contrast, immune function is bolstered by short day lengths in some species. The specific mechanisms that link reproductive and immune functions remain unspecified. Leptin is a hormone produced by adipose tissue, and several studies suggest that leptin modulates reproductive and immune functions. The present study sought to determine if photoperiodic alterations in reproductive function and leptin concentrations are linked to photoperiod modulated changes in immune function. Siberian hamsters (Phodopus sungorus) were housed in either long (LD 16:8) or short (LD 8:16) day lengths for 9 wk. After 9 wk, blood samples were collected during the middle of the light and dark phase to assess leptin concentrations. One week later, animals were injected with keyhole limpet hemocyanin to evaluate humoral immunity. Body mass, body fat content, and serum leptin concentrations were correlated with reproductive responsiveness to photoperiod; short-day animals with regressed gonads exhibited a reduction in these measures, whereas short-day nonresponders resembled long-day animals. In contrast, immune function was influenced by photoperiod but not reproductive status. Taken together, these data suggest that humoral immune function in Siberian hamsters is independent of photoperiod-mediated changes in leptin concentrations. PMID- 10848505 TI - Antipyretic effect of arginine vasotocin in toads. AB - Arginine vasotocin (AVT) is a nonmammalian analog of the mammalian hormone arginine vasopressin (AVP). These peptides are known for their antidiuretic and pressor effects. More recently, AVP has been recognized as an important antipyretic molecule in mammals. However, no information exists about the role of AVT in febrile ectotherms. We tested the hypothesis that AVT is an antipyretic molecule in the toad Bufo paracnemis. Toads equipped with a temperature probe were placed in a thermal gradient, and preferred body temperature was recorded continuously. A behavioral fever was observed after lipopolysaccharide (LPS) was injected systemically (200 microg/kg). Systemically injected AVT (300 pmol/kg) alone caused no significant change in body temperature, but abolished LPS-induced fever. Moreover, a smaller dose of AVT (10 pmol/kg), which did not affect LPS induced fever when injected peripherally, abolished fever when injected intracerebroventricularly. We therefore conclude that AVT plays an antipyretic role in the central nervous system, by means of behavior, in an ectotherm, a fact consistent with the notion that AVT/AVP elicits antipyresis by reducing the thermoregulatory set point. PMID- 10848506 TI - Inhibition of cyclooxygenase-2 prevents inflammation-mediated preterm labor in the mouse. AB - Prostaglandins (PGs) have proven important during parturition, but inhibition of PG production treating preterm labor (PTL) results in significant maternal and fetal side effects. We hypothesize that specific inhibition of either cyclooxygenase (COX)-1 or -2 may result in separation of therapeutic and toxic effects. We demonstrate that COX-2, but not COX-1, is induced during inflammation mediated PTL caused by lipopolysaccharide (LPS) administration. A two- to threefold increase in uterine and ovarian PG concentrations coincides with this induction of COX-2. The COX-2-selective inhibitor SC-236 proved effective in stopping preterm delivery and the increases in PGs. The COX-1-selective inhibitor SC-560 also attenuated uterine and ovarian PG production after LPS but did not inhibit PTL as efficiently as SC-236. COX-1-deficient mice, which show delay in the onset of term labor, exhibited no delay in onset of PTL after LPS. These findings suggest that the mechanisms for initiation of inflammation-mediated PTL and term labor differ and that selective COX-2 inhibition may provide a means of stopping inflammation-induced PTL in humans. PMID- 10848507 TI - Energy intake-independent modulation of triglyceride metabolism by glucocorticoids in the rat. AB - This study aimed to dissociate the peripheral effects of adrenalectomy (ADX) on triglyceride (TG) metabolism from those it exerts centrally on energy intake and to determine the impact of diet composition therein. Rats were fed either rodent chow or a diet high in sucrose and fat (HSF) and were adrenalectomized or left intact and pair fed to the ADX animals. Liver TG content, an index of hepatic TG production, was not affected by ADX, but was increased twofold by the HSF diet. ADX decreased the rate of hepatic TG secretion by 41% in chow-fed but not in HSF fed animals. Triglyceridemia and postheparin plasma lipase activities remained largely unchanged by treatments. ADX decreased insulinemia fivefold in chow-fed rats, but less so in HSF-fed animals. Likewise, subcutaneous and visceral adipose depots were 40-60% smaller in ADX than in intact pair-fed rats given chow, but the effect of ADX was dampened by consumption of the HSF diet. Although smaller, adipose tissues of ADX rats maintained a higher activity of lipoprotein lipase (LPL) than those of intact pair-fed rats, whereas muscle LPL was decreased. The study confirms that in the presence of reduced energy intake, corticosterone contributes to the maintenance of adipose stores and that the consequences of its absence tend to be attenuated when a high-energy diet is fed. The study further shows that, contrary to ad libitum feeding conditions, most determinants of TG metabolism, such as hepatic TG stores, triglyceridemia, postheparin plasma LPL, and adipose tissue LPL, are minimally affected by glucocorticoids when consumption of a high-energy diet is restricted, suggesting that glucocorticoids affect TG metabolism mostly indirectly through their central action on ingestive behavior. PMID- 10848508 TI - Baroreceptor afferent discharge in the pregnant rat. AB - Pregnancy is associated with blunted reflex responses to cardiac and arterial baroreceptor stimulation. We tested the hypothesis that arterial baroreceptor afferent discharge is attenuated in response to a pressure stimulus in pregnant rats. Multifiber aortic depressor nerve activity (ADNA), mean arterial pressure (MAP), and heart rate were measured in anesthetized (pentobarbital sodium, 35 mg/kg ip) late-pregnant and virgin rats in response to increases ?phenylephrine (PE), 1.5-24 microg. kg(-1). min(-1) and 1-16 microg/kg and decreases ?sodium nitroprusside (SNP), 5-80 microg. kg(-1). min(-1) and 0.05-16 microg/kg in MAP. Resting MAP was lower in pregnant rats, but changes in MAP were similar to those in virgin rats during both PE and SNP administration. ADNA was significantly attenuated in pregnant animals during both PE and SNP infusions (P < 0.05) due to a more rapid adaptation to the pressure stimulus. Bolus drug administration evoked similar changes in MAP and ADNA in both groups; however, the maximum decrease in ADNA was achieved at the lowest dose of SNP in pregnant rats. Thus baroreceptor afferent discharge is attenuated in pregnant rats, and this involves a more rapid adaptation to a pressure stimulus. PMID- 10848510 TI - Correlation structure of end-expiratory lung volume in anesthetized rats with intact upper airway. AB - The correlation structure of breath-to-breath fluctuations of end-expiratory lung volume (EEV) was studied in anesthetized rats with intact airways subjected to positive and negative transrespiratory pressure (i.e., PTRP and NTRP, correspondingly). The Hurst exponent, H, was estimated from EEV fluctuations using modified dispersional analysis. We found that H for EEV was 0.5362 +/- 0.0763 and 0.6403 +/- 0.0561 with PTRP and NTRP, respectively (mean +/- SD). Both H were significantly different from those obtained after random shuffling of the original time series. Also, H with NTRP was significantly greater than that with PTRP (P = 0.029). We conclude that in rats breathing through the upper airway, a positive long-term correlation is present in EEV that is different between PTRP and NTRP. PMID- 10848509 TI - Increased central AT(1)-receptor activation, not systemic vasopressin, sustains hypertension in ANP knockout mice. AB - We tested the hypothesis that hypertension in atrial natriuretic peptide (ANP) knockout mice is caused in part by disinhibition of angiotensin II-mediated vasopressin release. Inactin-anesthetized F(2) homozygous ANP gene-disrupted mice (-/-) and wild-type (+/+) littermates were surgically prepared for carotid arterial blood pressure measurement (ABP) and background intravenous injection of physiological saline or vasopressin V(1)-receptor antagonist (Manning compound, 10 ng/g body wt) and subsequent intracerebroventricular (left lateral ventricle) injection of saline (5 microl) or ANP (0.5 microg) or angiotensin II AT(1) receptor antagonist losartan (10 microg). Only (-/-) showed significant decrease in ABP after intracerebroventricular ANP or losartan. Both showed significant hypotension after intravenous V(1) antagonist, but there was no difference between their responses. We conclude that 1) vasopressin contributes equally to ABP maintenance in ANP-disrupted mice and wild-type controls; 2) permanently elevated ABP in ANP knockouts is associated with increased central nervous angiotensin II AT(1)-receptor activation; 3) disinhibition of central nervous angiotensin II AT(1) receptors in ANP-deficient animals does not lead to a significant increase in the importance of vasopressin as a mechanism for blood pressure maintenance. PMID- 10848511 TI - Ontogeny of neuronal nitric oxide synthase, NOS I, in the developing porcine kidney. AB - To determine if the developing kidney differs from the adult in the expression of the neuronal nitric oxide synthase, NOS I, these experiments measured mRNA gene expression by RNase protection assay and protein content by Western blot of NOS I in piglets at ages newborn and 3, 7, 10, 14, and 21 days and adult pigs. Whole kidney NOS I mRNA was greatest at birth and decreased progressively during renal maturation to adult levels. NOS I protein content paralleled this developmental pattern. Cortical NOS I protein was equivalent in newborn and 14-day-old piglets and was greater at both ages than the adult. Medullary NOS I protein was relatively greater than cortical in both immature ages and decreased from a peak at birth to adult levels. We conclude the following. 1) During postnatal maturation, renal NOS I mRNA and protein content show a pattern that is developmentally regulated. 2) This developmental pattern of NOS I after birth may, in part, contribute to the enhanced functional role of NO during renal maturation. PMID- 10848512 TI - Developmental changes in respiratory, febrile, and cardiovascular responses to PGE(2) in newborn lambs. AB - PGE(2) has centrally mediated respiratory, febrile, and cardiovascular effects that markedly differ between fetal and adult life. We hypothesized that the transition from fetal to adult responses to PGE(2) occurs in the newborn period. Thus effects of an intracarotid infusion of PGE(2) (3 microg/min for 60 min) were determined in unanesthetized newborn lambs at 5, 10, and 15 days after birth. At 5 days, PGE(2) reduced central CO(2) sensitivity, reduced lung ventilation due to a decrease in breathing frequency, and induced hypercapnia. By 15 days, these effects of PGE(2) had waned significantly. In contrast, phasic (expiratory) thyroarytenoid muscle electromyogram activity, number of short apneas, and incidence of Biot periodic breathing were similarly increased at all three ages. PGE(2) induced a sustained fever at 10 and 15 days. Heart rate and mean arterial blood pressure were unchanged in contrast to marked increases observed by others in adults. Results showed that the transition from fetal to adult respiratory and febrile responses to PGE(2) occurs in early postnatal life, whereas adult cardiovascular responses develop later in life in sheep. PMID- 10848513 TI - Central oxytocin modulates exercise-induced tachycardia. AB - We have shown that vasopressinergic projections to dorsal brain stem are activated during exercise and facilitate exercise tachycardia in both trained (T) and sedentary (S) rats (Dufloth DL, Morris M, and Michelini LC. Am J Physiol Regulatory Integrative Comp Physiol 273: R1271-R1282, 1997). In the present study, we investigated whether oxytocinergic projections to the nucleus of the solitary tract (NTS)-dorsal motor nucleus of the vagus (DMV) complex (NTS/DMV) are involved in the differential heart rate (HR) response to exercise in T and S rats. Arterial pressure (AP) and HR responses to dynamic exercise (0.4-1.4 km/h) were compared in S and T pretreated with vehicle (saline), oxytocin (OT; 20 pmol/200 nl) or OT-receptor antagonist (OT(ant); 20 pmol/200 nl) into the NTS/DMV. OT content in specific brain regions and plasma were measured in separate S and T groups at rest and immediately after exercise. Exercise increased OT content in dorsal (4.5-fold) and ventral brain stem (2.7-fold) and spinal cord (3.4-fold) only in T rats. No significant changes were observed in neurosecretory regions or medial eminence and posterior pituitary, but plasma levels of T rats were reduced immediately after exercise. Blockade of NTS/DMV OT receptors did not change basal mean AP (MAP) and HR or the MAP response to exercise. However, OT(ant) potentiated exercise-induced tachycardia (average increase of 26%) only in the T group. Pretreatment with exogenous OT in the NTS/DMV blunted the tachycardic response both in S and T rats without changing the MAP response. Administration of OT-receptor antagonist or OT into the fourth cerebral ventricle had no effect on the cardiovascular response to dynamic exercise. Taken together, the results suggest that oxytocinergic projections to the NTS/DMV are stimulated when T rats exercise and that OT released at this level acts on OT receptors to restrain exercise-induced tachycardia. PMID- 10848514 TI - Endogenous tachykinins cause bradycardia by stimulating cholinergic neurons in the isolated guinea pig heart. AB - The purpose of this study was to determine if endogenous tachykinins can cause bradycardia in the isolated perfused guinea pig heart through stimulation of cholinergic neurons. Capsaicin was used to stimulate release of tachykinins and calcitonin gene-related peptide (CGRP) from cardiac afferents. A bolus injection of 100 nmol capsaicin increased heart rate by 26 +/- 7% from a baseline of 257 +/ 14 beats/min (n = 6, P < 0.01). This positive chronotropic response was converted to a minor bradycardic effect in hearts with 1 microM CGRP-(8-37) present to block CGRP receptors. The negative chronotropic response to capsaicin was markedly potentiated in another group of hearts with the further addition of 0.5 microM neostigmine to inhibit cholinesterases. In this group, capsaicin decreased heart rate by 30 +/- 10% from a baseline of 214 +/- 6 beats/min (n = 8, P < 0.05). This large bradycardic response to capsaicin was inhibited by 1) infusion of neurokinin A to desensitize tachykinin receptors or 2) treatment with 1 microM atropine to block muscarinic receptors. The latter observations implicate tachykinins and acetylcholine, respectively, as mediators of the bradycardia. These findings support the hypothesis that endogenous tachykinins could mediate axon reflexes to stimulate cholinergic neurons of the intrinsic cardiac ganglia. PMID- 10848515 TI - Circadian rhythm of plasma sodium is disrupted in spontaneously hypertensive rats fed a high-NaCl diet. AB - High-NaCl diets elevate arterial pressure in NaCl-sensitive individuals, and increases in plasma sodium may trigger this effect. The present study tests the hypotheses that 1) plasma sodium displays a circadian rhythm in rats, 2) the plasma sodium rhythm is disturbed in spontaneously hypertensive rats (SHR), and 3) excess dietary NaCl elevates plasma sodium concentration in SHR. The results demonstrate that plasma sodium has a circadian rhythm that is inversely related to the circadian rhythm of arterial pressure. Although the plasma sodium rhythms of SHR and control rats are nearly identical, the plasma sodium concentrations are significantly higher in SHR throughout the 24-h cycle. Maintenance on a high NaCl diet increases plasma sodium concentration similarly in both SHR and control rats, but it blunts the plasma sodium rhythm only in SHR. These results demonstrate that in rats, plasma sodium has a circadian rhythm and that high-NaCl diets increase plasma sodium concentration. PMID- 10848516 TI - Cyclooxygenase-2 inhibitors constrict the fetal lamb ductus arteriosus both in vitro and in vivo. AB - Nonselective cyclooxygenase (COX) inhibitors are potent tocolytic agents; however, they also have adverse fetal effects such as constriction of the fetal ductus arteriosus. Recently, selective COX-2 inhibitors have been used in the management of preterm labor in the hope of avoiding fetal complications. However, both COX-1 and -2 are expressed by cells of the ductus arteriosus. We used fetal lambs (0.88 gestation) to assess the ability of selective COX-2 inhibitors celecoxib and NS398 to affect the ductus arteriosus. Both selective COX-2 inhibitors decreased PGE(2) and 6ketoPGF(1alpha) production in vitro; both inhibitors constricted the isolated ductus in vitro. The nonselective COX-1/COX-2 inhibitor indomethacin produced a further reduction in PG release and an additional increase in ductus tension in vitro. We used a prodrug of celecoxib to achieve 1.4 +/- 0.6 microg/ml, mean +/- standard deviation, of the active drug in vivo. This concentration of celecoxib produced both an increase in pressure gradient and resistance across the ductus; celecoxib also decreased fetal plasma concentrations of PGE(2) and 6ketoPGF(1alpha). Indomethacin (0.7 +/- 0.2 microg/ml) produced a significantly greater fall in ductus blood flow than celecoxib and tended to have a greater effect on ductus resistence in vivo. We conclude that caution should be used when recommending COX-2 inhibitors for use in pregnant women, because COX-2 appears to play a significant role in maintaining patency of the fetal ductus arteriosus. PMID- 10848517 TI - Characterization of L-arginine transporters in rat renal inner medullary collecting duct. AB - Previous work from our laboratory has demonstrated that the inner medullary collecting duct (IMCD) expresses a large amount of nitric oxide synthase (NOS) activity. The present study was designed to characterize the transport of NOS substrate, L-arginine, in a suspension of bulk-isolated IMCD cells from the Sprague-Dawley rat kidney. Biochemical transport studies demonstrated an L arginine transport system in IMCD cells that was saturable and Na(+) independent (n = 6). L-Arginine uptake by IMCD cells was inhibited by the cationic amino acids L-lysine, L-homoarginine, and L-ornithine (10 mmol/l each) and unaffected by the neutral amino acids L-leucine, L-serine, and L-glutamine. Both L-ornithine (n = 6) and L-lysine (n = 6) inhibited NOS enzymatic activity in a dose-dependent manner in IMCD cells, supporting the important role of L-arginine transport for NO production by this tubular segment. Furthermore, RT-PCR of microdissected IMCD confirmed the presence of cationic amino acid transporter CAT1 mRNA, whereas CAT2A, CAT2B, and CAT3 were not detected. These results indicate that L-arginine uptake by IMCD cells occurs via system y(+), is encoded by CAT1, and may participate in the regulation of NO production in this renal segment. PMID- 10848518 TI - Inhibitory effect of gurmarin on palatal taste responses to amino acids in the rat. AB - Gurmarin (10 microg/ml), a protein extracted from Gymnema sylvestre, depressed significantly (40-50%) the phasic taste responses to sugars (sucrose, fructose, lactose, and maltose) and saccharin sodium recorded from the greater superficial petrosal nerve (GSP) innervating palatal taste buds in the rat. However, no significant effect of gurmarin was observed for taste responses to NaCl, HCl, and quinine hydrochloride. Phasic responses to D-amino acids that taste sweet to humans (His, Asn, Phe, Gln) were also depressed, but gurmarin treatment was without significant effect on taste responses to D-Trp and D-Ala, six L-amino acids (His, Asn, Phe, Gln, Trp, and Ala), and two basic amino acid HCl salts (Arg and Lys). With the exception of D-Trp, these inhibitory effects of gurmarin on GSP taste responses were related to the rat's preference for these substances. PMID- 10848519 TI - Effects of leptin and cholecystokinin in rats with a null mutation of the leptin receptor Lepr(fak). AB - The Koletsky ("corpulent) obese rat is homozygous for an autosomal recessive mutation of the leptin receptor (Lepr) that results in hyperphagia, obesity, and hyperlipidemia. Unlike the Lepr mutation that characterizes the fatty Zucker rat (Lepr(fa)), the Koletsky mutation (Lepr(fak)) is null. Because the Lepr(fak) mutation is null, exogenous leptin should have no effect on body weight or food intake in fa(k)/fa(k) rats. We confirmed that prediction: murine leptin, administered into the third ventricle for 5 consecutive days, did not affect daily food intake or body weight in fa(k)/fa(k) rats but produced dose-related inhibitions of food intake and body weight in +/+ and +/fa(k) rats. Although fa(k)/fa(k) rats did not respond to leptin, their response to CCK-8 (4 microg/kg ip) injected before 30-min test meals of 10% sucrose was not different from that of +/+ or +/fa(k) rats. These results demonstrate that the fa(k)/fa(k) rat is a good model in which to analyze the controls of food intake, energy expenditure, and energy storage in the absence of leptin effects. PMID- 10848520 TI - Voltage-dependent calcium channels in ventricular cells of rainbow trout: effect of temperature changes in vitro. AB - Voltage-dependent calcium channels (VDCC) in ventricular myocytes from rainbow trout (Oncorhynchus mykiss) were investigated in vitro using the perforated patch clamp technique, which maintains the integrity of the intracellular milieu. First, we characterized the current using barium as the charge carrier and established the doses of various pharmacological agents to use these agents in additional studies. Second, we examined the current at several physiological temperatures to determine temperature dependency. The calcium currents at 10 degrees C (acclimation temperature) were identified as L-type calcium currents based on their kinetic behavior and response to various calcium channel agonists and antagonists. Myocytes were chilled (4 degrees C) and warmed (18 and 22 degrees C), and the response of VDCC to varying temperatures was observed. There was no significant dependency of the current amplitude and kinetics on temperature. Amplitude decreased 25-36% at 4 degrees C (Q(10) approximately 1.89) and increased 18% at 18 degrees C (Q(10) approximately 1.23) in control, Bay K8644 (Bay K)-, and forskolin-enhanced currents. The inactivation rates (tau(i)) did not demonstrate a temperature sensitivity for the VDCC (Q(10) 1.23-1. 92); Bay K treatment, however, increased temperature sensitivity of tau(i) between 10 and 18 degrees C (Q(10) 3.98). The low Q(10) values for VDCC are consistent with a minimal temperature sensitivity of trout myocytes between 4 and 22 degrees C. This low-temperature dependency may provide an important role for sarcolemmal calcium channels in adaptation to varying environmental temperatures in trout. PMID- 10848521 TI - Effects of temperature and calcium availability on ventricular myocardium from rainbow trout. AB - We studied the mechanical and electrophysiological properties of ventricular myocardium from rainbow trout (Oncorhynchus mykiss) in vitro at 4, 10, and 18 degrees C from fish acclimated at 10 degrees C. Temperature alone did not significantly alter the contractile force of the myocardium, but the time to peak tension and time to 80% relaxation were prolonged at 4 degrees C and shortened at 18 degrees C. The duration of the action potential was also prolonged at 4 degrees C and progressively shortened at higher temperatures. An alteration of the stimulation frequency did not affect contraction amplitude at any temperature. Calcium influx via L-type calcium channels was increased by raising extracellular calcium concentration (?Ca(2+)(o)) or including Bay K 8644 (Bay K) and isoproterenol in the bathing medium. These treatments significantly enhanced the contractile force at all temperatures. Calcium channel blockers had a reverse negative inotropic effect. Unexpectedly, the duration of the action potential at 10 degrees C was shortened as ?Ca(2+)(o) increased. However, Bay K prolonged the plateau phase at 4 degrees C. Caffeine, which promotes the release of sarcoplasmic reticulum (SR) calcium, increased contractile force eightfold at all three temperatures, but the SR blocker ryanodine was only inhibitory at 4 degrees C. Our results suggest that contractile force in ventricular myocardium from Oncorhynchus mykiss is primarily regulated by sarcolemmal calcium influx and that ventricular contractility is maintained during exposure to a wide range of temperatures. PMID- 10848522 TI - Effects of thyroid hormone receptor gene disruption on myosin isoform expression in mouse skeletal muscles. AB - Skeletal muscle is known to be a target for the active metabolite of thyroid hormone, i.e., 3,5,3'-triiodothyronine (T(3)). T(3) acts by repressing or activating genes coding for different myosin heavy chain (MHC) isoforms via T(3) receptors (TRs). The diverse function of T(3) is presumed to be mediated by TR alpha(1) and TR-beta, but the function of specific TRs in regulating MHC isoform expression has remained undefined. In this study, TR-deficient mice were used to expand our knowledge of the mechanisms by which T(3) regulates the expression of specific MHC isoforms via distinct TRs. In fast-twitch extensor digitorum longus (EDL) muscle, TR-alpha(1)-, TR-beta-, or TR-alpha(1)beta-deficient mice showed a small but statistically significant decrease (P < 0.05) of type IIB MHC content and an increased number of type I fibers. In the slow-twitch soleus, the beta/slow MHC (type I) isoform was significantly (P < 0. 001) upregulated in the TR-deficient mice, but this effect was highly dependent on the type of receptor deleted. The lack of TR-beta had no significant effect on the expression of MHC isoforms. An increase (P < 0.05) of type I MHC was observed in the TR-alpha(1) deficient muscle. A dramatic overexpression (P < 0.001) of the slow type I MHC and a corresponding downregulation of the fast type IIA MHC (P < 0.001) was observed in TR-alpha(1)beta-deficient mice. The muscle- and fiber-specific differences in MHC isoform expression in the TR-alpha(1)beta-deficient mice resembled the MHC isoform transitions reported in hypothyroid animals, i.e., a mild MHC transition in the EDL, a dramatic but not complete upregulation of the beta/slow MHC isoform in the soleus, and a variable response to TR deficiency in different soleus muscle fibers. Thus the consequences on muscle are similar in the absence of thyroid hormone or absence of thyroid hormone receptors, indicating that TR-alpha(1) and TR-beta together mediate the known actions of T(3). However, it remains unknown how thyroid hormone exerts muscle- and muscle fiber-specific effects in its action. Finally, although developmental MHC transitions were not studied specifically in this study, the absence of embryonic and fetal MHC isoforms in the TR-deficient mice indicates that ultimately the transition to the adult MHC isoforms is not solely mediated by TRs. PMID- 10848523 TI - Hormonal regulation of expression of ileal bile acid binding protein in suckling rats. AB - Ileal bile acid binding protein (IBABP) is a cytosolic protein believed to be involved in the absorption of conjugated bile acids. In rodents this protein and its mRNA have been shown to increase markedly during the third postnatal week. Because this period of ontogeny is characterized by increasing circulating concentrations of glucocorticoids and thyroxine, the goal of our study was to investigate the role of these hormones in IBABP expression in the developing rat. Administration of various doses of dexamethasone (Dex) during the second postnatal week caused a robust induction of IBABP mRNA and protein. Plateau levels of IBABP mRNA occurred at a Dex dose of 0.1 microg/g body wt, which is within the physiological range. IBABP mRNA was not appreciably induced until 24 h after treatment, suggesting that glucocorticoids influence IBABP either through a delayed primary or a secondary response mechanism. The regional pattern of IBABP mRNA elicited by Dex mimicked that seen during normal development, with appearance in distal ileum preceding proximal ileum. Thyroxine injections did not result in a significant increase of IBABP mRNA, and synergism between Dex and thyroxine was not observed. Taken together, our data suggest that maturation of IBABP expression is influenced by glucocorticoids but not by thyroxine. PMID- 10848524 TI - Bone and shell contribution to lactic acid buffering of submerged turtles Chrysemys picta bellii at 3 degrees C. AB - To evaluate shell and bone buffering of lactic acid during acidosis at 3 degrees C, turtles were submerged in anoxic or aerated water and tested at intervals for blood acid-base status and plasma ions and for bone and shell percent water, percent ash, and concentrations of lactate, Ca(2+), Mg(2+), P(i), Na(+), and K(+). After 125 days, plasma lactate concentration rose from 1.6 +/- 0.2 mM (mean +/- SE) to 155.2 +/- 10.8 mM in the anoxic group but only to 25.2 +/- 6.4 mM in the aerated group. The acid-base state of the normoxic animals was stable after 25 days of submergence. Plasma calcium concentration (?Ca(2+)) rose during anoxia from 3.2 +/- 0.2 to 46.0 +/- 0.6 mM and ?Mg(2+) from 2.7 +/- 0.2 to 12.2 +/- 0.6 mM. Both shell and bone accumulated lactate to concentrations of 135.6 +/- 35.2 and 163.6 +/- 5.1 mmol/kg wet wt, respectively, after 125 days anoxia. Shell and bone ?Na(+) both fell during anoxia but the fate of this Na(+) is uncertain because plasma ?Na(+) also fell. No other shell ions changed significantly in concentration, although the concentrations of both bone calcium and bone potassium changed significantly. Control shell water (27.8 +/- 0.6%) was less than bone water (33.6 +/- 1.1%), but neither changed during submergence. Shell ash (44.7 +/- 0.8%) remained unchanged, but bone ash (41.0 +/- 1.0%) fell significantly. We conclude that bone, as well as shell, accumulate lactate when plasma lactate is elevated, and that both export sodium carbonate, as well as calcium and magnesium carbonates, to supplement ECF buffering. PMID- 10848526 TI - Metabolic inhibitors synergistically decrease hepatic energy status and increase food intake. AB - Previous studies indicate that administration of the metabolic inhibitor, 2,5 anhydro-D-mannitol (2,5-AM) or methyl palmoxirate (MP), induces feeding behavior in rats by lowering hepatic energy status. Combined treatment with these agents synergistically increases food intake. The present study was designed to investigate whether combined treatment also has a synergistic effect on hepatic energy status. Rats treated with both inhibitors increased feeding behavior compared with the controls, whereas those treated with 2, 5-AM or MP alone did not. Although 2,5-AM alone lowered hepatic ATP content regardless of MP treatment, only the combination resulted in decreases in hepatic ATP/ADP ratio and phosphorylation potential. MP treatment did not affect the uptake of 2,5-AM into liver. These results suggest that a reduction in hepatic energy status is the common triggering signal for eating behavior induced by 2,5-AM and MP and provide additional evidence for an integrated metabolic control of food intake. PMID- 10848525 TI - GnRH stimulates LH release directly via inositol phosphate and indirectly via cAMP in African catfish. AB - In African catfish, two gonadotropin-releasing hormone (GnRH) peptides have been identified: chicken GnRH (cGnRH)-II and catfish GnRH (cfGnRH). The GnRH receptors on pituitary cells producing gonadotropic hormone signal through inositol phosphate (IP) elevation followed by increases in intracellular calcium concentration (?Ca(2+)(i)). In primary pituitary cell cultures of male African catfish, both cGnRH-II and cfGnRH dose dependently elevated IP accumulation, ?Ca(2+)(i), and the release of the luteinizing hormone (LH)-like gonadotropin. In all cases, cGnRH-II was more potent than cfGnRH. The GnRH-stimulated LH release was not associated with elevated cAMP levels, and forskolin-induced cAMP elevation had no effect on LH release. With the use of pituitary tissue fragments, however, cAMP was elevated by GnRH, and forskolin was able to stimulate LH secretion. Incubating these fragments with antibodies against cfGnRH abolished the forskolin-induced LH release but did not compromise the forskolin induced cAMP elevation. This suggests that cfGnRH-containing nerve terminals are present in pituitary tissue fragments and release cfGnRH via cAMP signaling on GnRH stimulation, whereas the GnRH receptors on gonadotrophs use IP/?Ca(2+)(i) to stimulate the release of LH. PMID- 10848527 TI - Arterial pulse pressure and vasopressin release during graded water immersion in humans. AB - Previous results indicate that arterial pulse pressure modulates release of arginine vasopressin (AVP) in humans. The hypothesis was therefore tested that an increase in arterial pulse pressure is the stimulus for suppression of AVP release during central blood volume expansion by water immersion. A two-step immersion model (n = 8) to the xiphoid process and neck, respectively, was used to attain two different levels of augmented cardiac distension. Left atrial diameter (echocardiography) increased from 28 +/- 1 to 34 +/- 1 mm (P < 0.05) during immersion to the xiphoid process and more so (P < 0.05), to 36 +/- 1 mm, during immersion to the neck. During immersion to the xiphoid process, arterial pulse pressure (invasively measured in a brachial artery) increased (P < 0.05) from 44 +/- 1 to 51 +/- 2 mmHg and to the same extent from 42 +/- 1 to 52 +/- 2 mmHg during immersion to the neck. Mean arterial pressure was unchanged during immersion to the xiphoid process and increased during immersion to the neck by 7 +/- 1 mmHg (P < 0.05). Arterial plasma AVP decreased from 2.5 +/- 0.7 to 1.8 +/- 0.5 pg/ml (P < 0. 05) during immersion to the xiphoid process and significantly more so (P < 0.05), to 1.4 +/- 0.5 pg/ml, during immersion to the neck. In conclusion, other factors besides the increase in arterial pulse pressure must have participated in the graded suppression of AVP release, comparing immersion to the xiphoid process with immersion to the neck. We suggest that when arterial pulse pressure is increased, graded distension of cardiopulmonary receptors modulate AVP release. PMID- 10848528 TI - Neural control of intestinal ion transport and paracellular permeability is altered by nutritional status. AB - This study examined the effect of fasting on the neural control of ion transport and paracellular permeability in piglet jejunum. Muscle-stripped tissues from fed or 48-h fasted piglets were mounted in Ussing chambers. Neural blockade with tetrodotoxin (TTX) or antagonists of muscarinic or nicotinic receptors caused reductions in basal short-circuit current that were approximately threefold greater in fasted piglets. The TTX-induced reduction in short-circuit current in fasted piglets was due to a decrease in residual ion flux and was abolished in the absence of HCO(-)(3). Intestinal paracellular permeability, as indicated by tissue conductance (G(t)) and fluxes of inulin and mannitol, was significantly increased by fasting. TTX increased inulin flux and G(t) in fed but not fasted piglets. In fasted piglets, carbachol reduced G(t) by 29% and mannitol flux by 27% but had no effect on these parameters in the fed state. We conclude that fasting enhances enteric neural control of basal ion transport and increases paracellular permeability in piglet jejunum. Tonic release of enteric neurotransmitters regulates paracellular permeability in the fed state, and cholinergic stimulation restores fasting-induced elevations in paracellular permeability to fed levels. PMID- 10848529 TI - Subthreshold aortic nerve inputs to neurons in nucleus of the solitary tract. AB - Subthreshold aortic nerve (AN) inputs to neurons receiving a monosynaptic AN evoked input (MSNs: respond to each of two AN stimuli separated by 5 ms) and neurons receiving a polysynaptic AN input (PSNs) in the nucleus of the solitary tract (NTS) were identified in anesthetized rats. In extracellular recordings from 24 MSNs and 49 PSNs, 12% of MSNs and 29% of PSNs only responded to AN stimulation during the application of excitatory amino acids. In intracellular recordings from 24 MSNs and 22 PSNs, 12% of MSNs and 14% of PSNs responded to AN stimulation with excitatory postsynaptic potentials that did not evoke action potential discharge. Reductions in arterial pressure produced minimal changes in the spontaneous discharge of suprathreshold AN-evoked neurons, suggesting that these neurons receive excitatory inputs from nonbaroreceptor sources. The results suggest that some baroreflex-related NTS neurons exist in a "reserve state and can be changed to an active state or vice versa. This will change the number of neurons involved in baroreflex circuits and provides a novel mechanism for regulating baroreflex function independently of alterations in peripheral afferent input. PMID- 10848530 TI - Effects of right atrial distension on the activity of magnocellular neurons in the supraoptic nucleus. AB - A small balloon placed at the junction of the superior vena cava and right atrium was used to stimulate cardiac volume receptors in pentobarbital sodium anesthetized male rats. Extracellular recordings were obtained from antidromically identified vasopressinergic and oxytocinergic neurosecretory cells of the supraoptic nucleus. Cells were considered sensitive to the stimulus if balloon inflation resulted in a 30% change in firing frequency. Balloon inflation that did not stretch the caval-atrial junction had no significant effect on vasopressin neurons (n = 51, P > 0.05). Stretch of the caval-atrial junction decreased the firing activity in 64 of 83 putative vasopressin neurons (P < 0.01 compared with control). Stretch of the caval-atrial junction influenced the firing activity of only 3 of 26 antidromically activated oxytocinergic neurons, an effect not statistically different from control (P > 0. 05). When bilateral vagotomy was performed while recording from vasopressin neurons (n = 5), sensitivity to stretch of the caval-atrial junction was eliminated. Cardiac receptors located at the junction of the superior vena cava and right atrium may be important in regulating the activity of vasopressinergic but not oxytocinergic neurons of the supraoptic nucleus. PMID- 10848531 TI - Defenselike patterns of spinal sympathetic outflow involving the 10-Hz and cardiac-related rhythms. AB - Frequency- and time-domain analyses were used to compare the effects of stimulation of the defense region of the midbrain periaqueductal gray (PAG) on the 10-Hz and cardiac-related discharges of sympathetic nerves with different cardiovascular targets. In baroreceptor-denervated cats anesthetized with urethan, PAG stimulation at frequencies equal to or higher (up to 25 Hz) than that of the free-running 10-Hz rhythm produced an immediate and sustained decrease in vertebral sympathetic nerve (VN) 10-Hz activity but increased the 10 Hz discharges of the inferior cardiac (CN) and renal (RN) nerves. In baroreceptor innervated cats, VN cardiac-related activity was initially unchanged by high frequency (25-Hz) PAG stimulation, or it increased along with that in the CN and RN. Later, during high-frequency PAG stimulation, when the rise in blood pressure approached its peak, VN cardiac-related activity usually was reduced below control level. At this time, the increases in CN and RN cardiac-related discharges were largely sustained. The cardiac-related discharges of the three nerves were unaffected by PAG stimulation at frequencies just below or just above that of the heartbeat. We conclude that the defenselike pattern of spinal sympathetic outflow involving the 10-Hz rhythm is different in mechanism and character from that involving the cardiac-related rhythm. PMID- 10848532 TI - NPY-leptin: opposing effects on appetitive and consummatory ingestive behavior and sexual behavior. AB - Many studies have indicated that neuropeptide Y (NPY) stimulates and leptin inhibits food intake. In line with this, intracerebroventricular injection of NPY (10 microg) stimulated and leptin (10 microg) inhibited intake of a sucrose solution when female rats were required to obtain the solution from a bottle. However, NPY inhibited and leptin stimulated intake if the solution was infused intraorally. Thus NPY stimulates the responses used to obtain food but inhibits those used to consume food, and leptin has the opposite effects. To test the specificity of these responses the sexual behavior of male rats was examined. NPY treated males showed minor deficits in sexual behavior but chose to ingest a sucrose solution rather than copulate with a female if offered the choice. By contrast, leptin-treated males ingested little sucrose and displayed an increase in ejaculatory frequency if given the same choice. It is suggested that NPY is not merely an orexigenic peptide, but one that directs attention toward food. Similarly, leptin may not be an anorexic peptide, but one that diverts attention away from food toward alternate stimuli. PMID- 10848533 TI - Arterial baroreflex regulation of regional vascular conductance at rest and during exercise. AB - We tested the hypothesis that dynamic exercise resets the operating point and attenuates the spontaneous gain of the arterial baroreflex regulation of mesenteric and hindlimb vascular conductance in hypertensive rats. Eleven adult male spontaneously hypertensive rats were chronically instrumented with left carotid arterial catheters and Doppler ultrasonic flow probes around the superior mesenteric and left common iliac arteries. After the rats recovered, arterial baroreflex function was examined by recording reflex changes in conductance in response to spontaneous changes in mean arterial pressure before exercise and during steady-state treadmill running at 6 and 18 m/min. Dynamic exercise reduced the spontaneous baroreflex gain of mesenteric conductance (by 51 and 36%) and maximum mesenteric conductance (by 24 and 32%) at 6 and 18 m/min, respectively. In sharp contrast, dynamic exercise increased the spontaneous maximum iliac conductance (by 32 and 47%) without changing the spontaneous gain. Sinoaortic denervation eliminated the relationship between mean arterial pressure and conductance by reducing the mesenteric (92%) and iliac (68%) vascular conductance gain. These results demonstrate that dynamic exercise has differential effects on the regulation of mesenteric and iliac vascular conductance in hypertensive rats. PMID- 10848534 TI - A projection from the ventral tegmental area to the periaqueductal gray involved in cardiovascular regulation. AB - Experiments were done in alpha-chloralose-anesthetized rats to determine a pathway mediating the cardiovascular depressor responses elicited from stimulation of the ventral tegmental area (VTA). The magnitude of the depressor responses elicited by glutamate stimulation (0.1 M/30 nl) of the VTA was examined after neuronal block produced by microinjections of lidocaine into ascending fiber bundles leaving the VTA to innervate the forebrain and thalamus. Bilateral microinjections of 1 microl of 4% lidocaine in the medial forebrain bundle (n = 6) and in the periventricular fibers of the midbrain (n = 5) did not attenuate the depressor response from stimulation of the VTA. Experiments were done using the anterograde tracer biotinylated dextran amine to identify descending projections from the VTA to cardiovascular centers in the brain stem. Examination of the nucleus of the solitary tract, ventrolateral medulla, and A5 catecholaminergic cell group revealed few or no fibers or terminals. Occasional fibers and some terminals were observed in the nucleus of raphe magnus, parabrachial nucleus, and locus ceruleus. A very dense bilateral projection was found to the ventrolateral periaqueductal gray (PAGvl) and dorsal raphe nucleus adjacent to the PAGvl. Bilateral injections of 4% lidocaine (n = 4) or 10 mM cobalt chloride (n = 5) into the PAGvl region attenuated the depressor responses elicited by stimulation of the VTA by approximately 50%. These experiments indicate that the depressor responses elicited from activation of the VTA are mediated in part by a pathway to a cardiovascular depressor area located in the PAGvl. PMID- 10848535 TI - Mitogenic whey extract stimulates wound repair activity in vitro and promotes healing of rat incisional wounds. AB - The ability of single growth factors to promote healing of normal and compromised wounds has been well described, but wound healing is a process requiring the coordinated action of multiple growth factors. Only the synergistic effect on wound healing of combinations containing at most two individual growth factors has been reported. We sought to assess the ability of a novel milk-derived growth factor-enriched preparation ?mitogenic bovine whey extract (MBWE), which contains six known growth factors, to promote repair processes in organotypic in vitro models and incisional wounds in vivo. MBWE stimulated the contraction of fibroblast-populated collagen lattices in a dose-dependent fashion and promoted the closure of excisional wounds in embryonic day 17 fetal rat skin. Application of MBWE increased incisional wound strength in normal animals on days 3, 5, 7, and 10 and reversed the decrease in wound strength observed following steroid treatment. Wound histology showed increased fibroblast numbers in wounds from normal and steroid-compromised animals. These data suggest the mixture of factors present in bovine milk exerts a direct action on the cells of cutaneous wound repair to enhance both normal and compromised healing. PMID- 10848536 TI - Is the spring quality of muscle plastic? AB - During locomotion, major muscle groups are often activated cyclically. This alternate stretch-shorten pattern of activity could enable muscle to function as a spring, storing and recovering elastic recoil potential energy. Because the ability to store and recover elastic recoil energy could profoundly affect the energetics of locomotion, one might expect this to be an adaptable feature of skeletal muscle. This study tests the hypothesis that chronic eccentric (Ecc) training results in a change in the spring properties of skeletal muscle. Nine female Sprague-Dawley rats underwent chronic Ecc training for 8 wk on a motorized treadmill. The spring properties of muscle were characterized by both active and passive lengthening force productions. A single "spring constant (Deltaforce/Deltalength) from the passive length-tension curves was calculated for each muscle. Results from measurements on long heads of triceps brachii muscle indicate that the trained group produced significantly more passive lengthening force (P = 0.0001) as well as more active lengthening force (P = 0.0001) at all lengths of muscle stretch. In addition, the spring constants were significantly different between the Ecc (1.71 N/mm) and the control (1.31 N/mm) groups. A stiffer spring is capable of storing more energy per unit length stretched, which is of functional importance during locomotion. PMID- 10848537 TI - Hypercapnia-induced cerebral and ocular vasodilation is not altered by glibenclamide in humans. AB - Carbon dioxide is an important regulator of vascular tone. Glibenclamide, an inhibitor of ATP-sensitive potassium channel (K(ATP)) activation, significantly blunts vasodilation in response to hypercapnic acidosis in animals. We investigated whether glibenclamide also alters the cerebral and ocular vasodilator response to hypercapnia in humans. Ten healthy male subjects were studied in a controlled, randomized, double-blind two-way crossover study under normoxic and hypercapnic conditions. Glibenclamide (5 mg po) or insulin (0.3 mU. kg(-1). min(-1) iv) were administered with glucose to achieve comparable plasma insulin levels. In control experiments, five healthy volunteers received glibenclamide (5 mg) or nicorandil (40 mg) or glibenclamide and nicorandil in a randomized, three-way crossover study. Mean blood flow velocity and resistive index in the middle cerebral artery (MCA) and in the ophthalmic artery (OA) were measured with Doppler sonography. Pulsatile choroidal blood flow was assessed with laser interferometric measurement of fundus pulsation. Forearm blood flow was measured with venous occlusion plethysmography. Hypercapnia increased ocular fundus pulsation amplitude by +18.2-22.3% (P < 0. 001) and mean flow velocity in the MCA by +27.4-33.3% (P < 0.001), but not in the OA (2.1-6.5%, P = 0.2). Forearm blood flow increased by 78.2% vs. baseline (P = 0.041) after nicorandil administration. Glibenclamide did not alter hypercapnia-induced changes in cerebral or ocular hemodynamics and did not affect systemic hemodynamics or forearm blood flow but significantly increased glucose utilization and blunted the nicorandil-induced vasodilation in the forearm. This suggests that hypercapnia-induced changes in the vascular beds under study are not mediated by activation of K(ATP) channels in humans. PMID- 10848538 TI - Hepatic versus gallbladder bile composition: in vivo transport physiology of the gallbladder in rainbow trout. AB - Ion and water transport across the teleost Oncorhynchus mykiss gallbladder were studied in vivo by comparing flow and composition of hepatic bile, collected by chronic catheter, to volume and composition of terminally collected gallbladder bile. Differences in composition were comparable with those of other vertebrates, whereas bile flow (75 microl. kg(-1). h(-1)) was below values reported for endothermic vertebrates. The gallbladder concentrates bile acids five- to sevenfold and exhibits higher net Cl(-) than Na(+) transport in vivo, in contrast to the 1:1 transport ratio from gallbladders under saline/saline conditions. Transepithelial potential (TEP) in the presence of bile, at the apical surface, was -13 mV (bile side negative) but +1.5 mV in the presence of saline. Bile acid in the apical saline reversed the TEP, presumably by a Donnan effect. We propose that ion transport across the gallbladder in vivo involves backflux of Na(+) from blood to bile resulting in higher net Cl(-) than Na(+) flux. This Na(+) backflux is driven by a bile side negative TEP and low Na(+) activity in bile due to the complexing effects of bile acids. PMID- 10848539 TI - Evidence for an intrarenal renin-angiotensin system in the rainbow trout, Oncorhynchus mykiss. AB - Physiological and molecular approaches were used to investigate the existence of an intrarenal renin-angiotensin system (RAS) in rainbow trout. Inhibition of angiotensin-converting enzyme by captopril (5 x 10(-4 )M) rapidly decreased vascular resistance of the trunk of the trout, perfused at 19 mmHg, resulting in an increased perfusate flow rate and a decreased intrarenal dorsal aortic pressure. A profound diuresis occurred in the in situ perfused kidney and reflected both increased glomerular filtration rates and decreased water reabsorption (osmolyte reabsorption was unchanged). Renal and vascular parameters recovered once captopril treatment was stopped. Diuretic and vascular effects of captopril on the in situ trout kidney concur with an inhibition of known vasoconstrictor and antidiuretic actions of angiotensin II. However, at a higher perfusion pressure (28 mmHg), captopril had no effect on intrarenal aortic pressure or perfusate and urine flow rates, suggesting that the trout intrarenal RAS is activated by low perfusion pressures/flows. Existence of the renal RAS in trout was further supported by evidence for angiotensinogen gene expression in kidney as well as liver. PMID- 10848540 TI - Cooperation of muscle and cutaneous afferents in the feedback of contraction to peroneal motoneurons. AB - Peroneal motoneurons were recorded intracellularly in anesthetized cats during sustained submaximal contractions of peroneus brevis muscle (PB) elicited by repetitive electrical stimulation of motor axons in the distal portion of cut ventral root filaments. Mechanical stimulation of the territory innervated by the superficial peroneal nerve (SP) was applied during contraction to assess the influence of afferents from this territory on the contraction-induced excitation of motoneurons. In 21 peroneal motoneurons in which PB contraction evoked excitatory potentials, a stimulation engaging mechanoreceptors located in the skin around toes was found to either enhance (in 12 motoneurons) or reduce (in 9 motoneurons) the contraction-induced excitatory potentials. Among positive effects, six showed simple summation of the responses to each individual stimulus, suggesting a convergence of afferent pathways on motoneurons. In six other motoneurons, complex interactions were observed, as may result from convergence at a premotoneuronal level. Among negative effects, a single instance was observed of inhibitory facilitation, as may result from convergence of cutaneous and muscular, possibly Ib, afferents on inhibitory interneurons. Several pathways, mediating either facilitory or inhibitory influences, are available for cooperation of muscle and cutaneous input, allowing flexibility of motoneuron activation in different tasks. PMID- 10848541 TI - Effects on peroneal motoneurons of cutaneous afferents activated by mechanical or electrical stimulations. AB - The postsynaptic potentials elicited in peroneal motoneurons by either mechanical stimulation of cutaneous areas innervated by the superficial peroneal nerve (SP) or repetitive electrical stimulation of SP were compared in anesthetized cats. After denervation of the foot sparing only the territory of SP terminal branches, reproducible mechanical stimulations were applied by pressure on the plantar surface of the toes via a plastic disk attached to a servo-length device, causing a mild compression of toes. This stimulus evoked small but consistent postsynaptic potentials in every peroneal motoneuron. Weak stimuli elicited only excitatory postsynaptic potentials (EPSPs), whereas increase in stimulation strength allowed distinction of three patterns of response. In about one half of the sample, mechanical stimulation or trains of 20/s electric pulses at strengths up to six times the threshold of the most excitable fibers in the nerve evoked only EPSPs. Responses to electrical stimulation appeared with 3-7 ms central latencies, suggesting oligosynaptic pathways. In another, smaller fraction of the sample, inhibitory postsynaptic potentials (IPSPs) appeared with an increase of stimulation strength, and the last fraction showed a mixed pattern of excitation and inhibition. In 24 of 32 motoneurons where electrical and mechanical effects could be compared, the responses were similar, and in 6 others, they changed from pure excitation on mechanical stimulation to mixed on electrical stimulation. With both kinds of stimulation, stronger stimulations were required to evoke inhibitory postsynaptic potentials (IPSPs), which appeared at longer central latencies than EPSPs, indicating longer interneuronal pathways. The similarity of responses to mechanical and electrical stimulation in a majority of peroneal motoneurons suggests that the effects of commonly used electrical stimulation are good predictors of the responses of peroneal motoneurons to natural skin stimulation. The different types of responses to cutaneous afferents from SP territory reflect a complex connectivity allowing modulations of cutaneous reflex responses in various postures and gaits. PMID- 10848542 TI - Light-evoked excitatory synaptic currents of X-type retinal ganglion cells. AB - The excitatory amino acid receptor (EAAR) types involved in the generation of light-evoked excitatory postsynaptic currents (EPSCs) were examined in X-type retinal ganglion cells. Using isolated and sliced preparations of cat and ferret retina, the light-evoked EPSCs of X cells were isolated by adding picrotoxin and strychnine to the bath to remove synaptic inhibition. N-methyl-D-aspartate (NMDA) receptors contribute significantly to the light-evoked EPSCs of ON- and OFF-X cells at many different holding potentials. An NMDA receptor contribution to the EPSCs was observable when retinal synaptic inhibition was either normally present or pharmacologically blocked. NMDA receptors formed 80% of the peak light-evoked EPSC at a holding potential of -40 mV; however, even at -80 mV, 20% of the light evoked EPSC was NMDA-mediated. An alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptor-mediated component to the light-evoked EPSCs predominated at a holding potential of -80 mV. The light-evoked EPSC was blocked by the AMPA receptor-selective antagonist GYKI52466 (50-100 microM). The AMPA receptor-mediated EPSC component had a linear current-voltage relation. AMPA receptors form the main non-NMDA EAAR current on both ON- and OFF- X ganglion cell dendrites. When synaptic transmission was blocked by the addition of Cd(2+) to the Ringer, application of kainate directly to ganglion cells evoked excitatory currents that were strongly blocked by GYKI52466. Experiments using selective EAAR modulators showed the AMPA receptor-selective modulator cyclothiazide potentiated glutamate-evoked currents on X cells, while the kainate receptor-selective modulator concanavalin A (ConA) had no effect on kainate evoked currents. Whereas the present study confirms the general notion that AMPA EAAR-mediated currents are transient and NMDA receptor-mediated currents are sustained, current-voltage relations of the light-evoked EPSC at different time points showed the contributions of these two receptor types significantly overlap. Both NMDA and AMPA EAARs can transmit transient and sustained visual signals in X ganglion cells, suggesting that much signal shaping occurs presynaptically in bipolar cells. PMID- 10848543 TI - Reaching during virtual rotation: context specific compensations for expected coriolis forces. AB - Subjects who are in an enclosed chamber rotating at constant velocity feel physically stationary but make errors when pointing to targets. Reaching paths and endpoints are deviated in the direction of the transient inertial Coriolis forces generated by their arm movements. By contrast, reaching movements made during natural, voluntary torso rotation seem to be accurate, and subjects are unaware of the Coriolis forces generated by their movements. This pattern suggests that the motor plan for reaching movements uses a representation of body motion to prepare compensations for impending self-generated accelerative loads on the arm. If so, stationary subjects who are experiencing illusory self rotation should make reaching errors when pointing to a target. These errors should be in the direction opposite the Coriolis accelerations their arm movements would generate if they were actually rotating. To determine whether such compensations exist, we had subjects in four experiments make visually open loop reaches to targets while they were experiencing compelling illusory self rotation and displacement induced by rotation of a complex, natural visual scene. The paths and endpoints of their initial reaching movements were significantly displaced leftward during counterclockwise illusory rotary displacement and rightward during clockwise illusory self-displacement. Subjects reached in a curvilinear path to the wrong place. These reaching errors were opposite in direction to the Coriolis forces that would have been generated by their arm movements during actual torso rotation. The magnitude of path curvature and endpoint errors increased as the speed of illusory self-rotation increased. In successive reaches, movement paths became straighter and endpoints more accurate despite the absence of visual error feedback or tactile feedback about target location. When subjects were again presented a stationary scene, their initial reaches were indistinguishable from pre-exposure baseline, indicating a total absence of aftereffects. These experiments demonstrate that the nervous system automatically compensates in a context-specific fashion for the Coriolis forces associated with reaching movements. PMID- 10848544 TI - Saccades from torsional offset positions back to listing's plane. AB - Rapid eye movements include saccades and quick phases of nystagmus and may have components around all three axes of ocular rotation: horizontal, vertical, and torsional. In this study, we recorded horizontal, vertical, and torsional eye movements in normal subjects with their heads upright and stationary. We asked how the eyes are brought back to Listing's plane after they are displaced from it. We found that torsional offsets, induced with a rotating optokinetic disk oriented perpendicular to the subject's straight ahead, were corrected during both horizontal and vertical voluntary saccades. Thus three-dimensional errors are synchronously reduced during saccades. The speed of the torsional correction was much faster than could be accounted for by passive mechanical forces. During vertical saccades, the peak torsional velocity decreased and the time of peak torsional velocity was delayed, as the amplitude of vertical saccades increased. In contrast, there was no consistent reduction of torsional velocity or change in time of peak torsional velocity with an increase in the amplitude of horizontal saccades. These findings suggest that 1) the correction of stimulus-induced torsion is neurally commanded and 2) there is cross-coupling between the torsional and vertical but not between the torsional and horizontal saccade generating systems. This latter dichotomy may reflect the fact that vertical and torsional rapid eye movements are generated by common premotor circuits located in the rostral interstitial nucleus of the medial longitudinal fasciculus (riMLF). When horizontal or vertical saccade duration was relatively short, the torsional offset was not completely corrected during the primary saccade, indicating that although the saccade itself is three-dimensional, saccade duration is determined by the error in the horizontal or the vertical, but not by the error in the torsional component. PMID- 10848545 TI - Nicotinic EPSCs in intact rat ganglia feature depression except if evoked during intermittent postsynaptic depolarization. AB - The involvement of the postsynaptic membrane potential level in controlling synaptic strength at the ganglionic synapse was studied by recording nicotinic fast synaptic currents (EPSCs) from neurons in the intact, mature rat superior cervical ganglion, using the two-electrode voltage-clamp technique. EPSCs were evoked by 0.05-Hz supramaximal stimulation of the preganglionic sympathetic trunk over long periods; their peak amplitude (or synaptic charge transfer) over time appeared to depend on the potential level of the neuronal membrane where the nicotinic receptors are embedded. EPSC amplitude remained constant (n = 6) only if ACh was released within repeated depolarizing steps of the postganglionic neuron, which constantly varied between -50 and -20 mV in consecutive 10-mV steps, whereas it decreased progressively by 45% (n = 9) within 14 min when the sympathetic neuron was held at constant membrane potential. Synaptic channel activation, channel ionic permeation and depolarization of the membrane in which the nicotinic receptor is localized must occur simultaneously to maintain constant synaptic strength at the ganglionic synapse during low-rate stimulation (0.03-1 Hz). Different posttetanic (20 Hz for 10 s) behaviors were observed depending on the mode of previous stimulation. In the neuron maintained at constant holding potential during low-rate stimulation, the depressed EPSC showed posttetanic potentiation, recovering approximately 23% of the mean pretetanic values (n = 10). The maximum effect was immediate in 40% of the neurons tested and developed over a 3- to 6-min period in the others; thereafter potentiation vanished within 40 min of 0.05-Hz stimulation. In contrast, no statistically significant synaptic potentiation was observed when EPSC amplitudes were kept constant by repeated -50/-20-mV command cycles (n = 12). It is suggested that, under these conditions, posttetanic potentiation could represent an attempt at recovering the synaptic strength lost during inappropriate functioning of the ganglionic synapse. PMID- 10848546 TI - Three-dimensional vestibuloocular reflex of the monkey: optimal retinal image stabilization versus listing's law. AB - If the rotational vestibuloocular reflex (VOR) were to achieve optimal retinal image stabilization during head rotations in three-dimensional space, it must turn the eye around the same axis as the head, with equal velocity but in the opposite direction. This optimal VOR strategy implies that the position of the eye in the orbit must not affect the VOR. However, if the VOR were to follow Listing's law, then the slow-phase eye rotation axis should tilt as a function of current eye position. We trained animals to fixate visual targets placed straight ahead or 20 degrees up, down, left or right while being oscillated in yaw, pitch, and roll at 0.5-4 Hz, either with or without a full-field visual background. Our main result was that the visually assisted VOR of normal monkeys invariantly rotated the eye around the same axis as the head during yaw, pitch, and roll (optimal VOR). In the absence of a visual background, eccentric eye positions evoked small axis tilts of slow phases in normal animals. Under the same visual condition, a prominent effect of eye position was found during roll but not during pitch or yaw in animals with low torsional and vertical gains following plugging of the vertical semicircular canals. This result was in accordance with a model incorporating a specific compromise between an optimal VOR and a VOR that perfectly obeys Listing's law. We conclude that the visually assisted VOR of the normal monkey optimally stabilizes foveal as well as peripheral retinal images. The finding of optimal VOR performance challenges a dominant role of plant mechanics and supports the notion of noncommutative operations in the oculomotor control system. PMID- 10848547 TI - Bovine versus rat adrenal chromaffin cells: big differences in BK potassium channel properties. AB - Both bovine and rat adrenal chromaffin cells have served as pioneering model systems in cellular neurophysiology, including in the study of large conductance calcium- and voltage-dependent K(+) (BK) channels. We now report that while BK channels dominate the outward current profile of both species, specific gating properties vary widely across cell populations, and the distributions of these properties differ dramatically between species. Although BK channels were first described in bovine chromaffin cells, rapidly inactivating ones were discovered in rat chromaffin cells. We report that bovine cells can also exhibit inactivating BK channels with varying properties similar to those in rat cells. However, a much smaller proportion of bovine cells exhibit inactivating BK current, the proportion of the total current that inactivates is usually smaller, and the rate of inactivation is often much slower. Other gating features differ as well; the voltage dependence of channel activation is much more positive for bovine cells, and their rates of activation and deactivation are faster and slower, respectively. Modeling studies suggest that channel heterogeneity is consistent with varying tetrameric combinations of inactivation-competent versus incompetent subunits. The results suggest that chromaffin BK channel functional nuances represent an important level for evolutionary tailoring of autonomic stress responses. PMID- 10848548 TI - Cannabinoids modulate synaptic strength and plasticity at glutamatergic synapses of rat prefrontal cortex pyramidal neurons. AB - Cannabinoids receptors have been reported to modulate synaptic transmission in many structures of the CNS, but yet little is known about their role in the prefrontal cortex where type I cannabinoid receptor (CB-1) are expressed. In this study, we tested first the acute effects of selective agonists and antagonist of CB-1 on glutamatergic excitatory postsynaptic currents (EPSCs) in slices of rat prefrontal cortex (PFC). EPSCs were evoked in patch-clamped layer V pyramidal cells by stimulation of layer V afferents. Monosynaptic EPSCs were strongly depressed by bath application (1 microM) of the cannabinoid receptors agonists WIN55212-2 (-50.4 +/- 8.8%) and CP55940 (-42.4 +/- 10.9%). The CB-1 antagonist SR141716A reversed these effects. Unexpectedly, SR141716A alone produced a significant increase of glutamatergic synaptic transmission (+46.9 +/- 11.2%), which could be partly reversed by WIN55212-2. In the presence of strontium in the bath, the frequency but not the amplitude of asynchronous synaptic events evoked in layer V pyramidal cells by stimulating layer V afferents, was markedly decreased (-54.2 +/- 8%), indicating a presynaptic site of action of cannabinoids at these synapses. Tetanic stimulation (100 pulses at 100 Hz, 4 trains) induced in control condition, no changes (n = 7/18), long-term depression (LTD; n = 6/18), or long-term potentiation (LTP; n = 5/18) of monosynaptic EPSCs evoked by stimulation of layer V afferents. When tetanus was applied in the presence of WIN 55,212-2 or SR141716-A (1 microM) in the bath, the proportion of "nonplastic" cells were not significantly changed (n = 7/15 in both cases). For the plastic ones (n = 8 in both cases), WIN 55,212-2 strongly favored LTD (n = 7/8) at the apparent expense of LTP (n = 1/8), whereas the opposite effect was observed with SR141716-A (7/8 LTP; 1/8 LTD). These results demonstrate that cannabinoids influence glutamatergic synaptic transmission and plasticity in the PFC of rodent. PMID- 10848549 TI - Prolonged synaptic integration in perirhinal cortical neurons. AB - Layer II/III of rat perirhinal cortex (PR) contains numerous late-spiking (LS) pyramidal neurons. When injected with a depolarizing current step, these LS cells typically delay spiking for one or more seconds from the onset of the current step and then sustain firing for the duration of the step. This pattern of delayed and sustained firing suggested a specific computational role for LS cells in temporal learning. This hypothesis predicts and requires that some layer II/III neurons should also exhibit delayed and sustained spiking in response to a train of excitatory synaptic inputs. Here we tested this prediction using visually guided, whole cell recordings from rat PR brain slices. Most LS cells (19 of 26) exhibited delayed spiking to synaptic stimulation (>1 s latency from the train onset), and the majority of these cells (13 of 19) also showed sustained firing that persisted for the duration of the synaptic train (5-10 s duration). Delayed and sustained firing in response to long synaptic trains has not been previously reported in vertebrate neurons. The data are consistent with our model that a circuit containing late spiking neurons can be used for encoding long time intervals during associative learning. PMID- 10848550 TI - Anabolic steroids induce region- and subunit-specific rapid modulation of GABA(A) receptor-mediated currents in the rat forebrain. AB - Anabolic-androgenic steroids (AAS) have become significant drugs of abuse in recent years with the highest increase reported in adolescent girls. In spite of the increased use of AAS, the CNS effects of these steroids are poorly understood. We report that in prepubertal female rats, three commonly abused AAS, 17alpha-methyltestosterone, stanozolol, and nandrolone, induced rapid and reversible modulation of GABAergic currents in neurons of two brain regions known to be critical for the expression of reproductive behaviors: the ventromedial nucleus of the hypothalamus (VMN) and the medial preoptic area (mPOA). All three AAS significantly enhanced peak synaptic current amplitudes and prolonged synaptic current decays in neurons of the VMN. Conversely all three AAS significantly diminished peak current amplitudes of synaptic currents from neurons of the mPOA. The endogenous neuroactive steroids, 3alpha-hydroxy-5alpha pregnan-20-one and 5alpha-androstane-3alpha,17beta-diol, potentiated currents in the VMN as did the AAS. In contrast to the negative modulation induced by AAS in the mPOA, the endogenous steroids potentiated responses in this region. To determine the concentration response relationships, modulation by the AAS, 17alpha-methyltestosterone (17alpha-meT), was assessed for currents evoked by ultrafast perfusion of brief pulses of GABA to acutely isolated neurons. Half maximal effects on currents elicited by 1 mM GABA were elicited by submicromolar concentrations of AAS for neurons from both brain regions. In addition, the efficacy of 10(-5) to 10(-2) M GABA was significantly increased by 1 microM 17alpha-meT. Previous studies have demonstrated a striking dichotomy in receptor composition between the VMN and the mPOA with regard to gamma subunit expression. To determine if the preferential expression of gamma(2) subunit-containing receptors in the VMN and of gamma(1) subunit-containing receptors in the mPOA could account for the region-specific effects of AAS in the two regions, responses elicited by ultrafast perfusion of GABA to human embryonic kidney 293 cells transfected with alpha(2), beta(3), and gamma(2) or alpha(2), beta(3), and gamma(1) subunit cDNAs were analyzed. As with native VMN neurons, positive modulation of GABA responses was elicited for alpha(2)beta(3)gamma(2) recombinant receptors, while negative modulation was induced at alpha(2)beta(3)gamma(1) receptors as in the mPOA. Our data demonstrate that AAS in doses believed to occur in steroid abusers can induce significant modulation of GABAergic transmission in brain regions essential for neuroendocrine function. In addition, the effects of these steroids can vary significantly between brain regions in a manner that appears to depend on the subunit composition of GABA(A) receptors expressed. PMID- 10848551 TI - Linear to supralinear summation of AMPA-mediated EPSPs in neocortical pyramidal neurons. AB - It has been hypothesized that voltage-sensitive conductances present on the dendrites of neurons can influence summation of excitatory postsynaptic potentials (EPSPs) and hence affect how neurons compile information. Greater than linear summation of EPSPs has been postulated to facilitate coincidence detection by cortical neurons. This study examined whether the summation of subthreshold AMPA-mediated EPSPs generated on layer V neocortical pyramidal neurons in vitro was linear and if any nonlinearities could be attributed to dendritic conductances. Evoked EPSPs (1-12 mV) were recorded somatically by means of intracellular sharp electrodes in the presence of 100 microM amino-5 phosphonopentanoic acid (AP-5) and 3 microM bicuculline. Two independent EPSPs were evoked by a stimulating electrode in layer I and another in layers III-V. The areas of stimulation were isolated from each other by a horizontal cut below layer I. By subtracting the algebraic sum of the individual EPSPs from the evoked response when both EPSPs were evoked simultaneously, we determined that they summed linearly to supralinearly. Supralinear summation was more likely when the soma was hyperpolarized by DC current injection. Summation was predominantly linear when postsynaptic conductances (i.e., Na(+) and Ca(2+)) were blocked with intracellular QX-314. The supralinear summation of EPSPs (without QX-314) decreased as the time between inputs was increased from 0 to 30 ms. To determine the role of dendrites in nonlinear summation, we substituted a current pulse (simulated EPSP) delivered at the soma for either or both of the evoked EPSPs. Simulated EPSPs combined with either an evoked EPSP or another simulated EPSP showed significantly less supralinear summation than two evoked EPSPs, indicating that the dendritic conductances were largely responsible for the observed supralinear summation. PMID- 10848552 TI - Multi-joint coordination during walking and foothold searching in the Blaberus cockroach. I. Kinematics and electromyograms. AB - Cockroaches were induced to walk or search for a foothold while they were tethered above a glass plate made slick with microtome oil. We combined kinematic analysis of leg joint movements with electromyographic (EMG) recordings from leg extensor muscles during tethered walking and searching to characterize these behaviors. The tethered preparation provides technical advantages for multi-joint kinematic and neural analysis. However, the behavioral relevance of the tethered preparation is an important issue. To address this issue, we evaluated the effects of tethering the animals by comparing kinematic parameters of tethered walking with similar data collected previously from cockroaches walking freely on a treadmill at the same speeds. No significant differences between tethered and treadmill walking were found for most joint kinematic parameters. In contrast, comparison of tethered walking and searching showed that the two behaviors can be distinguished by analysis of kinematics and electrical data. We combined analysis of joint kinematics and electromyograms to examine the change in multi-joint coordination during walking and searching. During searching, middle leg joints extended during swing rather than stance (i.e., walking) and the coordination of movements and extensor motor neuron activity at the coxa-trochanteral and femur tibia joints differed significantly during walking and searching. We also found that the pattern of myographic activity in the middle leg during searching was similar to that in the front legs during walking. PMID- 10848553 TI - Multi-joint coordination during walking and foothold searching in the Blaberus cockroach. II. Extensor motor neuron pattern. AB - In a previous study, we combined joint kinematics and electromyograms (EMGs) to examine the change in the phase relationship of two principal leg joints during walking and searching. In this study, we recorded intracellularly from motor neurons in semi-intact behaving animals to examine mechanisms coordinating extension at these leg joints. In particular, we examined the change in the phase of the coxa-trochanter (CTr) and femur-tibia (FT) joint extension during walking and searching. In doing so, we discovered marked similarities in the activity of CTr and FT joint extensor motor neurons at the onset of extension during searching and at the end of stance during walking. The data suggest that the same interneurons may be involved in coordinating the CTr and FT extensor motor neurons during walking and searching. Previous studies in stick insects have suggested that extensor motor neuron activity during the stance phase of walking results from an increase in tonic excitation of the neuron leading to spiking that is periodically interrupted by centrally generated inhibition. However, the CTr and FT extensor motor neuron activity during walking consists of characteristic phasic modulations in motor neuron frequency within each step cycle. The phasic increases and decreases in extensor EMG frequency during stance are associated with kinematic events (i.e., foot set-down and joint cycle transitions) during walking. Sensory feedback associated with these events might be responsible for phasic modulation of the extensor motor neuron frequency. However, our data rule out the possibility that sensory cues resulting from foot set-down are responsible for a decline in CTr extensor activity that is characteristic of the Blaberus step cycle. Our data also suggest that both phasic excitation and inhibition contribute to extensor motor neuron activity during the stance phase of walking. PMID- 10848554 TI - Contralateral movement and extensor force generation alter flexion phase muscle coordination in pedaling. AB - The importance of bilateral sensorimotor signals in coordination of locomotion has been demonstrated in animals but is difficult to ascertain in humans due to confounding effects of mechanical transmission of forces between the legs (i.e., mechanical interleg coupling). In a previous pedaling study, by eliminating mechanical interleg coupling, we showed that muscle coordination of a unipedal task can be shaped by interlimb sensorimotor pathways. Interlimb neural pathways were shown to alter pedaling coordination as subjects pedaling unilaterally exhibited increased flexion-phase muscle activity compared with bilateral pedaling even though the task mechanics performed by the pedaling leg(s) in the unilateral and bilateral pedaling tasks were identical. To further examine the relationship between contralateral sensorimotor state and ipsilateral flexion phase muscle coordination during pedaling, subjects in this study pedaled with one leg while the contralateral leg either generated an extensor force or relaxed as a servomotor either held that leg stationary or moved it in antiphase with the pedaling leg. In the presence of contralateral extensor force generation, muscle activity in the pedaling leg during limb flexion was reduced. Integrated electromyographic activity of the pedaling-leg hamstring muscles (biceps femoris and semimembranosus) during flexion decreased by 25-30%, regardless of either the amplitude of force generated by the nonpedaling leg or whether the leg was stationary or moving. In contrast, rectus femoris and tibialis anterior activity during flexion decreased only when the contralateral leg generated high rhythmic force concomitant with leg movement. The results are consistent with a contralateral feedforward mechanism triggering flexion-phase hamstrings activity and a contralateral feedback mechanism modulating rectus femoris and tibialis anterior activity during flexion. Because only muscles that contribute to flexion as a secondary function were observed, it is impossible to know whether the modulatory effect also acts on primary, unifunctional, limb flexors or is specific to multifunctional muscles contributing to flexion. The influence of contralateral extensor-phase sensorimotor signals on ipsilateral flexion may reflect bilateral coupling of gain control mechanisms. More generally, these interlimb neural mechanisms may coordinate activity between muscles that perform antagonistic functions on opposite sides of the body. Because pedaling and walking share biomechanical and neuronal control features, these mechanisms may be operational in walking as well as pedaling. PMID- 10848555 TI - Dynorphin exerts both postsynaptic and presynaptic effects in the Globus pallidus of the rat. AB - The opioids contained in striato-pallidal axons are thought to play a significant role in motor control. We examined post- and presynaptic effects of the kappa (kappa)-receptor agonist dynorphin A (1-13) (DYN13) on the globus pallidus (GP) neurons in rat brain slice preparations using the whole cell recording method. DYN13 hyperpolarized and decreased the input resistance of approximately one quarter of neurons examined. All of these DYN13-sensitive neurons had medium sized somata, large aspiny dendrites and generated repetitive firing without strong accommodation. The hyperpolarization was blocked by barium and was independent of TTX and intracellular chloride levels. The hyperpolarization was also selectively blocked by the kappa-antagonist nor-binaltorphimine dihydrochloride but not by the mu- or delta-antagonists. These data suggested that DYN13 activates barium-sensitive potassium currents in some GP neurons. Low- and high-intensity stimulation of the neostriatum (Str) evoked long- and short latency GABAergic responses, respectively. Previous data suggested that the long- and the short-latency responses were due to activation of the striato-pallidal axons and the local collaterals of pallido-striatal axons, respectively. DYN13 diminished the amplitude of both the short- and long-latency GABAergic responses in all the neurons tested. The effects of DYN13 on GABAergic postsynaptic responses were also selectively blocked by a kappa-antagonist. To investigate whether the effects were pre- or postsynaptic, the effects of DYN13 on spontaneous inhibitory postsynaptic potentials (IPSPs) and TTX-independent miniature-inhibitory postsynaptic currents (IPSCs) were examined. DYN13 decreased the frequency, but not the amplitude, of spontaneous IPSCs and calcium-dependent miniature-IPSCs. However, DYN13 did not alter the cadmium-insensitive miniature IPSCs. These results suggested that DYN13 suppressed GABA release from presynaptic terminals. This possibility was tested using a paired-stimulation test. DYN13 reduced the probability of evoking IPSCs to the first stimulation and greatly increased the success probability to the second stimulus. The amplitude of successfully evoked IPSCs was not changed with DYN13. DYN13 did not affect the excitatory postsynaptic potentials (EPSPs) or the response to iontophoretically applied GABA and glutamate. Together, these results suggest that DYN released from striato-pallidal axons controls the activity of GP neurons 1) by directly hyperpolarizing a population of neurons and 2) by presynaptically inhibiting GABA release from striato-pallidal and intrapallidal terminals. PMID- 10848556 TI - Suppression of hindlimb inputs to S-I forelimb-stump representation of rats with neonatal forelimb removal: GABA receptor blockade and single-cell responses. AB - Neonatal forelimb removal in rats results in the development of inappropriate hindlimb inputs in the forelimb-stump representation of primary somatosensory cortex (S-I) that are revealed when GABA(A) and GABA(B) receptor activity are blocked. Experiments carried out to date have not made clear what information is being suppressed at the level of individual neurons. In this study, three potential ways in which GABA-mediated inhibition could suppress hindlimb expression in the S-I stump representation were evaluated: silencing S-I neurons with dual stump and hindlimb receptive fields, silencing neurons with receptive fields restricted to the hindlimb alone, and/or selective silencing of hindlimb inputs to neurons that normally express a stump receptive field only. These possibilities were tested using single-unit recording techniques to evaluate the receptive fields of S-I forelimb-stump neurons before, during, and after blockade of GABA receptors with bicuculline methiodide (for GABA(A)) and saclofen (for GABA(B)). Recordings were also made from normal rats for comparison. Of 92 neurons recorded from the S-I stump representation of neonatally amputated rats, only 2.2% had receptive fields that included the hindlimb prior to GABA receptor blockade. During GABA receptor blockade, 54.3% of these cells became responsive to the hindlimb, and in all but two cases, these same neurons also expressed a stump receptive field. Most of these cells (82.0%) expressed only stump receptive fields prior to GABA receptor blockade. In 71 neurons recorded from normal rats, only 5 became responsive to the hindlimb during GABA receptor blockade. GABA receptor blockade of cortical neurons, in both normal and neonatally amputated rats, resulted in significant enlargements of receptive fields as well as the emergence of receptive fields for neurons that were normally unresponsive. GABA receptor blockade also resulted in increases in both the spontaneous activity and response magnitudes of these neurons. These data support the conclusion that GABA mechanisms generally act to specifically suppress hindlimb inputs to S-I forelimb stump neurons that normally express a receptive field on the forelimb stump only. PMID- 10848557 TI - Mechanisms involved in tetanus-induced potentiation of fast IPSCs in rat hippocampal CA1 neurons. AB - In the present study, possible mechanisms involved in the tetanus-induced potentiation of gamma-aminobutyric acid-A (GABA-A) receptor-mediated inhibitory postsynaptic currents (IPSCs) were investigated using the whole cell voltage clamp technique on CA1 neurons in rat hippocampal slices. Stimulations (100 Hz) of the stratum radiatum, while voltage-clamping the membrane potential of neurons, induces a long-term potentiation (LTP) of evoked fast IPSCs while increasing the number but not the amplitude of spontaneous IPSCs (sIPSCs). The potentiation of fast IPSCs was input specific. During the period of IPSC potentiation, postsynaptic responses produced by 4,5,6,7-tetrahydroisoxazolo[5,4 c]pyridin-3-ol hydrochloride and baclofen, GABA-A and GABA-B agonists respectively, were not significantly different from control. CGP 36742, a GABA-B antagonist, blocked the induction of tetanus-induced potentiation of evoked and spontaneous IPSCs, while GTPgammaS, an activator of G proteins, substitution for GTP in the postsynaptic recording electrode did not occlude potentiation. Since GABA-B receptors work through G proteins, our results suggest that pre- but not postsynaptic GABA-B receptors are involved in the potentiation of fast IPSCs. A tetanus delivered when GABA-A responses were completely blocked by bicuculline suggests that GABA-A receptor activation during tetanus is not essential for the induction of potentiation. Rp-cAMPs, an antagonist of protein kinase A (PKA) activation, blocks the induction of potentiation of fast IPSCs. Forskolin, an activator of PKA, increases baseline evoked IPSCs as well as the number of sIPSCs, and a tetanic stimulation during this enhancement uncovers a long-term depression of the evoked IPSC. Sulfhydryl alkylating agents, N-ethylmaleimide and p-chloromercuribenzoic acid, which have been found to presynaptically increase GABA release and have been suggested to have effects on proteins involved in transmitter release processes occurring in nerve terminals, occlude tetanus induced potentiation of evoked and spontaneous IPSCs. Taken together our results suggest that LTP of IPSCs originates from a presynaptic site and that GABA-B receptor activation, cyclic AMP/PKA activation and sulfhydryl-alkylation are involved. Plasticity of IPSCs as observed in this study would have significant implications for network behavior in the hippocampus. PMID- 10848558 TI - Modeling attractor deformation in the rodent head-direction system. AB - We present a model of the head-direction circuit in the rat that improves on earlier models in several respects. First, it provides an account of some of the unique characteristics of head-direction (HD) cell firing in the lateral mammillary nucleus and the anterior thalamus. Second, the model functions without making physiologically unrealistic assumptions. In particular, it implements attractor dynamics in postsubiculum and lateral mammillary nucleus without directionally tuned inhibitory neurons, which have never been observed in vivo, and it integrates angular velocity without the use of multiplicative synapses. The model allows us to examine the relationships among three HD areas and various properties of their representations. A surprising result is that certain combinations of purported HD cell properties are mutually incompatible, suggesting that the lateral mammillary nucleus may not be the primary source of head direction input to anterior thalamic HD cells. PMID- 10848559 TI - Blink-perturbed saccades in monkey. I. Behavioral analysis. AB - Saccadic eye movements are thought to be influenced by blinking through premotor interactions, but it is still unclear how. The present paper describes the properties of blink-associated eye movements and quantifies the effect of reflex blinks on the latencies, metrics, and kinematics of saccades in the monkey. In particular, it is examined to what extent the saccadic system accounts for blink related perturbations of the saccade trajectory. Trigeminal reflex blinks were elicited near the onset of visually evoked saccades by means of air puffs directed on the eye. Reflex blinks were also evoked during a straight-ahead fixation task. Eye and eyelid movements were measured with the magnetic-induction technique. The data show that saccade latencies were reduced substantially when reflex blinks were evoked prior to the impending visual saccades as if these saccades were triggered by the blink. The evoked blinks also caused profound spatial-temporal perturbations of the saccades. Deflections of the saccade trajectory, usually upward, extended up to approximately 15 degrees. Saccade peak velocities were reduced, and a two- to threefold increase in saccade duration was typically observed. In general, these perturbations were largely compensated in saccade mid-flight, despite the absence of visual feedback, yielding near-normal endpoint accuracies. Further analysis revealed that blink-perturbed saccades could not be described as a linear superposition of a pure blink-associated eye movement and an unperturbed saccade. When evoked during straight-ahead fixation, blinks were accompanied by initially upward and slightly abducting eye rotations of approximately 2-15 degrees. Back and forth wiggles of the eye were frequently seen; but in many cases the return movement was incomplete. Rather than drifting back to its starting position, the eye then maintained its eccentric orbital position until a downward corrective saccade toward the fixation spot followed. Blink-associated eye movements were quite rapid, albeit slower than saccades, and the velocity-amplitude-duration characteristics of the initial excursions as well as the return movements were approximately linear. These data strongly support the idea that blinks interfere with the saccade premotor circuit, presumably upstream from the neural eye-position integrator. They also indicated that a neural mechanism, rather than passive elastic restoring forces within the oculomotor plant, underlies the compensatory behavior. The tight latency coupling between saccades and blinks is consistent with an inhibition of omnipause neurons by the blink system, suggesting that the observed changes in saccade kinematics arise elsewhere in the saccadic premotor system. PMID- 10848560 TI - Blink-perturbed saccades in monkey. II. Superior colliculus activity. AB - Trigeminal reflex blinks evoked near the onset of a saccade cause profound spatial-temporal perturbations of the saccade that are typically compensated in mid-flight. This paper investigates the influence of reflex blinks on the discharge properties of saccade-related burst neurons (SRBNs) in intermediate and deep layers of the monkey superior colliculus (SC). Twenty-nine SRBNs, recorded in three monkeys, were tested in the blink-perturbation paradigm. We report that the air puff stimuli, used to elicit blinks, resulted in a short-latency ( approximately 10 ms) transient suppression of saccade-related SRBN activity. Shortly after this suppression (within 10-30 ms), all neurons resumed their activity, and their burst discharge then continued until the perturbed saccade ended near the extinguished target. This was found regardless whether the compensatory movement was into the cell's movement field or not. In the limited number of trials where no compensation occurred, the neurons typically stopped firing well before the end of the eye movement. Several aspects of the saccade related activity could be further quantified for 25 SRBNs. It appeared that 1) the increase in duration of the high-frequency burst was well correlated with the (two- to threefold) increase in duration of the perturbed movement. 2) The number of spikes in the burst for control and perturbed saccades was quite similar. On average, the number of spikes increased only 14%, whereas the mean firing rate in the burst decreased by 52%. 3) An identical number of spikes were obtained between control and perturbed responses when burst and postsaccadic activity were both included in the spike count. 4) The decrease of the mean firing rate in the burst was well correlated with the decrease in the velocity of perturbed saccades. 5) Monotonic relations between instantaneous firing rate and dynamic motor error were obtained for control responses but not for perturbed responses. And 6) the high-frequency burst of SRBNs with short-lead and long-lead presaccadic activity (also referred to as burst and buildup neurons, respectively) showed very similar features. Our findings show that blinking interacts with the saccade premotor system already at the level of the SC. The data also indicate that a neural mechanism, rather than passive elastic restoring forces within the oculomotor plant, underlies the compensation for blink-related perturbations. We propose that these interactions occur downstream from the motor SC and that the latter may encode the desired displacement vector of the eyes by sending an approximately fixed number of spikes to the brainstem saccadic burst generator. PMID- 10848561 TI - Properties of a calcium-activated K(+) current on interneurons in the developing rat hippocampus. AB - Calcium-activated potassium currents have an essential role in regulating excitability in a variety of neurons. Although it is well established that mature CA1 pyramidal neurons possess a Ca(2+)-activated K(+) conductance (I(K(Ca))) with early and late components, modulation by various endogenous neurotransmitters, and sensitivity to K(+) channel toxins, the properties of I(K(Ca)) on hippocampal interneurons (or immature CA1 pyramidal neurons) are relatively unknown. To address this problem, whole-cell voltage-clamp recordings were made from visually identified interneurons in stratum lacunosum-moleculare (L-M) and CA1 pyramidal cells in hippocampal slices from immature rats (P3-P25). A biphasic calcium activated K(+) tail current was elicited following a brief depolarization from the holding potential (-50 mV). Analysis of the kinetic properties of I(K(Ca)) suggests that an early current component differs between these two cell types. An early I(K(Ca)) with a large peak current amplitude (200.8 +/- 13.2 pA, mean +/- SE), slow time constant of decay (70.9 +/- 3.3 ms), and relatively rapid time to peak (within 15 ms) was observed on L-M interneurons (n = 88), whereas an early I(K(Ca)) with a small peak current amplitude (112.5 +/- 7.3 pA), a fast time constant of decay (39.4 +/- 1.6 ms), and a slower time-to-peak (within 26 ms) was observed on CA1 pyramidal neurons (n = 85). Removal of extracellular calcium or addition of inorganic Ca(2+) channel blockers (cadmium, nickel, or cobalt) was used to demonstrate the calcium dependence of these currents. Addition of norepinephrine, carbachol, and a variety of channel toxins (apamin, iberiotoxin, verruculogen, paxilline, penitrem A, and charybdotoxin) were used to further distinguish between I(K(Ca)) on these two hippocampal cell types. Verruculogen (100 nM), carbachol (100 microM), apamin (100 nM), TEA (1 mM), and iberiotoxin (50 nM) significantly reduced early I(K(Ca)) on CA1 pyramidal neurons; early I(K(Ca)) on L-M interneurons was inhibited by apamin and TEA. Combined with previous work showing that the firing properties of hippocampal interneurons and pyramidal cells differ, our kinetic and pharmacological data provide strong support for the hypothesis that different types of Ca(2+)-activated K(+) current are present on these two cell types. PMID- 10848562 TI - Differential fall in ATP accounts for effects of temperature on hypoxic damage in rat hippocampal slices. AB - Intracellular recordings, ATP and cytosolic calcium measurements from CA1 pyramidal cells in rat hippocampal slices were used to examine the mechanisms by which temperature alters hypoxic damage. Hypothermia (34 degrees C) preserved ATP (1.7 vs. 0.8 nM/mg) and improved electrophysiologic recovery of the CA1 neurons after hypoxia; 58% of the neurons subjected to 10 min of hypoxia (34 degrees C) recovered their resting and action potentials, while none of the neurons at 37 degrees C recovered. Increasing the glucose concentration from 4 to 6 mM during normothermic hypoxia improved ATP (1.3 vs. 0.8 nM/mg) and mimicked the effects of hypothermia; 67% of the neurons recovered their resting and action potentials. Hypothermia attenuated the membrane potential changes and the increase in intracellular Ca(2+) (212 vs. 384 nM) induced by hypoxia. Changing the glucose concentration in the artificial cerebrospinal fluid primarily affects ATP levels during hypoxia. Decreasing the glucose concentration from 4 to 2 mM during hypothermic hypoxia worsened ATP, cytosolic Ca(2+), and electrophysiologic recovery. Ten percent of the neurons subjected to 4 min of hypoxia at 40 degrees C recovered their resting and action potentials; this compared with 60% of the neurons subjected to 4 min of normothermic hypoxia. None of the neurons subjected to 10 min of hypoxia at 40 degrees C recovered their resting and action potentials. Hyperthermia (40 degrees C) worsens the electrophysiologic changes and induced a greater increase in intracellular Ca(2+) (538 vs. 384 nM) during hypoxia. Increasing the glucose concentration from 4 to 8 mM during 10 min of hyperthermic hypoxia improved ATP (1.4 vs. 0.6 nM/mg), Ca(2+) (267 vs. 538 nM), and electrophysiologic recovery (90 vs. 0%). Our results indicate that the changes in electrophysiologic recovery with temperature are primarily due to changes in ATP and that the changes in depolarization and Ca(2+) are secondary to these ATP changes. Both primary and secondary changes are important for explaining the improved electrophysiologic recovery with hypothermia. PMID- 10848563 TI - I4AA-Sensitive chloride current contributes to the center light responses of bipolar cells in the tiger salamander retina. AB - Light-evoked currents in depolarizing and hyperpolarizing bipolar cells (DBCs and HBCs) were recorded under voltage-clamp conditions in living retinal slices of the larval tiger salamander. Responses to illumination at the center of the DBCs' and HBCs' receptive fields were mediated by two postsynaptic currents: DeltaI(C), a glutamate-gated cation current with a reversal potential near 0 mV, and DeltaI(Cl), a chloride current with a reversal potential near -60 mV. In DBCs DeltaI(C) was suppressed by L-2-amino-4-phosphonobutyric acid (L-AP4), and in HBCs it was suppressed by 6,7-dinitroquinoxaline-2,3-dione (DNQX). In both DBCs and HBCs DeltaI(Cl) was suppressed by imidazole-4-acetic acid (I4AA), a GABA receptor agonist and GABA(C) receptor antagonist. In all DBCs and HBCs examined, 10 microM I4AA eliminated DeltaI(Cl) and the light-evoked current became predominately mediated by DeltaI(C). The addition of 20 microM L-AP4 to the DBCs or 50 microM DNQX to HBCs completely abolished DeltaI(C). Focal application of glutamate at the inner plexiform layer elicited chloride currents in bipolar cells by depolarizing amacrine cells that release GABA at synapses on bipolar cell axon terminals, and such glutamate-induced chloride currents in DBCs and HBCs could be reversibly blocked by 10 microM I4AA. These experiments suggest that the light-evoked, I4AA-sensitive chloride currents (DeltaI(Cl)) in DBCs and HBCs are mediated by narrow field GABAergic amacrine cells that activate GABA(C) receptors on bipolar cell axon terminals. Picrotoxin (200 microM) or (1,2,5,6 tetrahydropyridine-4yl) methyphosphinic acid (TPMPA) (2 other GABA(C) receptor antagonists) did not block (but enhanced and broadened) the light-evoked DeltaI(Cl), although they decreased the chloride current induced by puff application of GABA or glutamate. The light response of narrow field amacrine cells were not affected by I4AA, but were substantially enhanced and broadened by picrotoxin. These results suggest that there are at least two types of GABA(C) receptors in bipolar cells: one exhibits stronger I4AA sensitivity than the other, but both can be partially blocked by picrotoxin. The GABA receptors in narrow field amacrine cells are I4AA insensitive and picrotoxin sensitive. The light-evoked DeltaI(Cl) in bipolar cells are mediated by the more strongly I4AA sensitive GABA(C) receptors. Picrotoxin, although acting as a partial GABA(C) receptor antagonist in bipolar cells, does not suppress DeltaI(Cl) because its presynaptic effects on amacrine cell light responses override its antagonistic postsynaptic actions. PMID- 10848564 TI - Manual tracking in two dimensions. AB - Manual tracking was studied by asking subjects to follow, with their finger, a target moving on a touch-sensitive video monitor. The target initially moved in a straight line at a constant speed and then, at a random point in time, made one abrupt change in direction. The results were approximated with a simple model according to which, after a reaction time, the hand moved in a straight line to intercept the target. Both the direction of hand motion and its peak speed could be predicted by assuming a constant time to intercept. This simple model was able to account for results obtained over a broad range of target speeds as well as the results of experiments in which both the speed and the direction of the target changed simultaneously. The results of an experiment in which the target acceleration was nonzero suggested that the error signals used during tracking are related to both speed and direction but poorly (if at all) to target acceleration. Finally, in an experiment in which target velocity remained constant along one axis but the perpendicular component underwent a step change, tracking along both axes was perturbed. This last finding demonstrates that tracking in two dimensions cannot be decomposed into its Cartesian components. However, an analytical model in a hand-centered frame of reference in which speed and direction are the controlled variables could account for much of the data. PMID- 10848565 TI - Development of potassium conductances in perinatal rat phrenic motoneurons. AB - Prior to the inception of inspiratory synaptic drive transmission from medullary respiratory centers, rat phrenic motoneurons (PMNs) have action potential and repetitive firing characteristics typical of immature embryonic motoneurons. During the period spanning from when respiratory bulbospinal and segmental afferent synaptic connections are formed at embryonic day 17 (E17) through to birth (gestational period is approximately 21 days), a pronounced transformation of PMN electrophysiological properties occurs. In this study, we test the hypothesis that the elaboration of action potential afterpotentials and the resulting changes in repetitive firing properties are due in large part to developmental changes in PMN potassium conductances. Ionic conductances were measured via whole cell patch recordings using a cervical slice-phrenic nerve preparation isolated from perinatal rats. Voltage- and current-clamp recordings revealed that PMNs expressed outward rectifier (I(KV)) and A-type potassium currents that regulated PMN action potential and repetitive firing properties throughout the perinatal period. There was an age-dependent leftward shift in the activation voltage and a decrease in the time-to-peak of I(KV) during the period from E16 through to birth. The most dramatic change during the perinatal period was the increase in calcium-activated potassium currents after the inception of inspiratory drive transmission at E17. Block of the maxi-type calcium-dependent potassium conductance caused a significant increase in action potential duration and a suppression of the fast afterhyperpolarizing potential. Block of the small conductance calcium-dependent potassium channels resulted in a marked suppression of the medium afterhyperpolarizing potential and an increase in the repetitive firing frequency. In conclusion, the increase in calcium-mediated potassium conductances are in large part responsible for the marked transformation in action potential shape and firing properties of PMNs from the time between the inception of fetal respiratory drive transmission and birth. PMID- 10848566 TI - Polarized synaptic interactions between intercalated neurons of the amygdala. AB - The intercalated (ITC) cell masses are small GABAergic cell clusters interposed between the basolateral (BL) complex and central (CE) nucleus of the amygdala. ITC cells receive excitatory afferents from the BL complex and generate feed forward inhibition in the CE nucleus. Recently it was shown that ITC cells could gate impulse traffic between the BL complex and CE nucleus in a spatiotemporally differentiated manner. In addition, it was hypothesized that lateromedial inhibitory interactions between different ITC cell clusters played a critical role in this respect. Given the potential importance of such conditional computations, the present study aimed to characterize the connectivity existing among ITC cells. To this end, whole cell recordings of ITC neurons were obtained under visual guidance in coronal slices of the guinea pig amygdala. Electrical stimuli applied in the BL complex primarily elicited excitatory responses when they were applied at the same lateromedial level or more medially than the recorded ITC cells. As the stimulation site was moved laterally, the character of the response shifted toward inhibition. Both bicuculline and non-N-methyl-D aspartate receptor antagonists abolished this BL-evoked inhibition, suggesting that it was not mediated by BL inhibitory cells projecting to ITC neurons. In keeping with this, local glutamate injections in and around the ITC clusters revealed that the most effective site to inhibit ITC cells were ITC clusters located laterally with respect to the recorded one. The activation of more medial ITC clusters evoked much smaller responses. Thus, connections between ITC clusters tend to run in a lateromedial direction. To identify the source of these directionally polarized synaptic interactions, the morphological features of ITC cells were analyzed by intracellular injection of Neurobiotin. This analysis revealed that the dendritic tree and axonal arbor of ITC cells are asymmetric in the lateromedial plane. In particular, their laterally directed dendrites were longer than the medial ones, whereas their laterally directed axon collaterals were shorter than the medial ones. It is concluded that the morphological asymmetry of ITC cells accounts for the directional polarization of inter-ITC connections. The significance of these findings for the gating of information transfer from the BL complex to the CE nucleus is discussed. PMID- 10848567 TI - Extracellular pH responses in CA1 and the dentate gyrus during electrical stimulation, seizure discharges, and spreading depression. AB - Since neuronal excitability is sensitive to changes in extracellular pH and there is regional diversity in the changes in extracellular pH during neuronal activity, we examined the activity-dependent extracellular pH changes in the CA1 region and the dentate gyrus. In vivo, in the CA1 region, recurrent epileptiform activity induced by stimulus trains, bicuculline, and kainic acid resulted in biphasic pH shifts, consisting of an initial extracellular alkalinization followed by a slower acidification. In vitro, stimulus trains also evoked biphasic pH shifts in the CA1 region. However, in CA1, seizure activity in vitro induced in the absence of synaptic transmission, by perfusing with 0 Ca(2+)/5 mM K(+) medium, was only associated with extracellular acidification. In the dentate gyrus in vivo, seizure activity induced by stimulation to the angular bundle or by injection of either bicuculline or kainic acid was only associated with extracellular acidification. In vitro, stimulus trains evoked only acidification. In the dentate gyrus in vitro, recurrent epileptiform activity induced in the absence of synaptic transmission by perfusion with 0 Ca(2+)/8 mM K(+) medium was associated with extracellular acidification. To test whether glial cell depolarization plays a role in the regulation of the extracellular pH, slices were perfused with 1 mM barium. Barium increased the amplitude of the initial alkalinization in CA1 and caused the appearance of alkalinization in the dentate gyrus. In both CA1 and the dentate gyrus in vitro, spreading depression was associated with biphasic pH shifts. These results demonstrate that activity dependent extracellular pH shifts differ between CA1 and dentate gyrus both in vivo and in vitro. The differences in pH fluctuations with neuronal activity might be a marker for the basis of the regional differences in seizure susceptibility between CA1 and the dentate gyrus. PMID- 10848568 TI - Task-related modulation of visual cortex. AB - We performed a series of experiments to quantify the effects of task performance on cortical activity in early visual areas. Functional magnetic resonance imaging (fMRI) was used to measure cortical activity in several cortical visual areas including primary visual cortex (V1) and the MT complex (MT+) as subjects performed a variety of threshold-level visual psychophysical tasks. Performing speed, direction, and contrast discrimination tasks produced strong modulations of cortical activity. For example, one experiment tested for selective modulations of MT+ activity as subjects alternated between performing contrast and speed discrimination tasks. MT+ responses modulated in phase with the periods of time during which subjects performed the speed discrimination task; that is, MT+ activity was higher during speed discrimination than during contrast discrimination. Task-related modulations were consistent across repeated measurements in each subject; however, significant individual differences were observed between subjects. Together, the results suggest 1) that specific changes in the cognitive/behavioral state of a subject can exert selective and reliable modulations of cortical activity in early visual cortex, even in V1; 2) that there are significant individual differences in these modulations; and 3) that visual areas and pathways that are highly sensitive to small changes in a given stimulus feature (such as contrast or speed) are selectively modulated during discrimination judgments on that feature. Increasing the gain of the relevant neuronal signals in this way may improve their signal-to-noise to help optimize task performance. PMID- 10848569 TI - Spinal cholinergic neurons activated during locomotion: localization and electrophysiological characterization. AB - The objective of the present study was to determine the location of the cholinergic neurons activated in the spinal cord of decerebrate cats during fictive locomotion. Locomotion was induced by stimulation of the mesencephalic locomotor region (MLR). After bouts of locomotion during a 7-9 h period, the animals were perfused and the L(3)-S(1) spinal cord segments removed. Cats in the control group were subjected to the same surgical procedures but no locomotor task. The tissues were sectioned and then stained by immunohistochemical methods for detection of the c-fos protein and choline acetyltransferase (ChAT) enzyme. The resultant c-fos labeling in the lumbar spinal cord was similar to that induced by fictive locomotion in the cat. ChAT-positive cells also clearly exhibited fictive locomotion induced c-fos labeling. Double labeling with c-fos and ChAT was observed in cells within ventral lamina VII, VIII, and possibly IX. Most of them were concentrated in the medial portion of lamina VII close to lamina X, similar in location to the partition and central canal cells found by Barber and collaborators. The number of ChAT and c-fos-labeled neurons was increased following fictive locomotion and was greatest in the intermediate gray, compared with dorsal and ventral regions. The results are consistent with the suggestion that cholinergic interneurons in the lumbar spinal cord are involved in the production of fictive locomotion. Cells in the regions positive for double labeled cells were targeted for electrophysiological study during locomotion, intracellular filling, and subsequent processing for ChAT immunohistochemistry. Three cells identified in this way were vigorously active during locomotion in phase with ipsilateral extension, and they projected to the contralateral side of the spinal cord. Thus a new population of spinal cord cells can be defined: cholinergic partition cells with commissural projections that are active during the extension phase of locomotion. PMID- 10848570 TI - Inaudible high-frequency sounds affect brain activity: hypersonic effect. AB - Although it is generally accepted that humans cannot perceive sounds in the frequency range above 20 kHz, the question of whether the existence of such "inaudible" high-frequency components may affect the acoustic perception of audible sounds remains unanswered. In this study, we used noninvasive physiological measurements of brain responses to provide evidence that sounds containing high-frequency components (HFCs) above the audible range significantly affect the brain activity of listeners. We used the gamelan music of Bali, which is extremely rich in HFCs with a nonstationary structure, as a natural sound source, dividing it into two components: an audible low-frequency component (LFC) below 22 kHz and an HFC above 22 kHz. Brain electrical activity and regional cerebral blood flow (rCBF) were measured as markers of neuronal activity while subjects were exposed to sounds with various combinations of LFCs and HFCs. None of the subjects recognized the HFC as sound when it was presented alone. Nevertheless, the power spectra of the alpha frequency range of the spontaneous electroencephalogram (alpha-EEG) recorded from the occipital region increased with statistical significance when the subjects were exposed to sound containing both an HFC and an LFC, compared with an otherwise identical sound from which the HFC was removed (i.e., LFC alone). In contrast, compared with the baseline, no enhancement of alpha-EEG was evident when either an HFC or an LFC was presented separately. Positron emission tomography measurements revealed that, when an HFC and an LFC were presented together, the rCBF in the brain stem and the left thalamus increased significantly compared with a sound lacking the HFC above 22 kHz but that was otherwise identical. Simultaneous EEG measurements showed that the power of occipital alpha-EEGs correlated significantly with the rCBF in the left thalamus. Psychological evaluation indicated that the subjects felt the sound containing an HFC to be more pleasant than the same sound lacking an HFC. These results suggest the existence of a previously unrecognized response to complex sound containing particular types of high frequencies above the audible range. We term this phenomenon the "hypersonic effect." PMID- 10848571 TI - Vestibular signals in the parasolitary nucleus. AB - Vestibular primary afferents project to secondary vestibular neurons located in the vestibular complex. Vestibular primary afferents also project to the uvula nodulus of the cerebellum where they terminate on granule cells. In this report we describe the physiological properties of neurons in a "new" vestibular nucleus, the parasolitary nucleus (Psol). This nucleus consists of 2,300 GABAergic neurons that project onto the ipsilateral inferior olive (beta-nucleus and dorsomedial cell column) as well as the nucleus reticularis gigantocellularis. These olivary neurons are the exclusive source of vestibularly modulated climbing fiber inputs to the cerebellum. We recorded the activity of Psol neurons during natural vestibular stimulation in anesthetized rabbits. The rabbits were placed in a three-axis rate table at the center of a large sphere, permitting vestibular and optokinetic stimulation. We recorded from 74 neurons in the Psol and from 23 neurons in the regions bordering Psol. The activity of 72/74 Psol neurons and 4/23 non-Psol neurons was modulated by vestibular stimulation in either the pitch or roll planes but not the horizontal plane. Psol neurons responded in phase with ipsilateral side-down head position or velocity during sinusoidal stimulation. Approximately 80% of the recorded Psol neurons responded to static roll-tilt. The optimal response planes of evoked vestibular responses were inferred from measurement of null planes. Optimal response planes usually were aligned with the anatomical orientation of one of the two ipsilateral vertical semicircular canals. The frequency dependence of null plane measurements indicated a convergence of vestibular information from otoliths and semicircular canals. None of the recorded neurons evinced optokinetic sensitivity. These results are consistent with the view that Psol neurons provide the vestibular signals to the inferior olive that eventually reached the cerebellum in the form of modulated climbing fiber discharges. These signals provide information about spatial orientation about the longitudinal axis. PMID- 10848572 TI - Endomorphin-1 and endomorphin-2 modulate responses of trigeminal neurons evoked by N-methyl-D-aspartic acid and somatosensory stimuli. AB - The present study investigated the modulation of N-methyl-D-aspartate (NMDA) evoked and peripheral cutaneous stimulus-evoked responses of trigeminal neurons by endomorphins, endogenous ligands for the mu-opioid receptor. Effects of endomorphins, administered microiontophoretically, were tested on the responses of nociceptive neurons recorded in the superficial and deeper dorsal horn of the medulla (trigeminal nucleus caudalis) in anesthetized rats. Endomorphin-1 and endomorphin-2 predominantly reduced the NMDA-evoked responses, producing an inhibitory effect of 54.1 +/- 2.96% (mean +/- SE; n = 34, P < 0.001) in 92% (34/37) of neurons and 63.6 +/- 3.61% (n = 32, P < 0.001) in 91% (32/35) of neurons, respectively. The inhibitory effect of endomorphins was modality specific; noxious stimulus-evoked responses were reduced more than nonnoxious stimulus-evoked responses. Naloxone applied at iontophoretic current that blocked the inhibitory effect of [D-Ala(2), N-Me-Phe(4), Gly(5)-ol]-enkephalin, reduced the peak inhibitory effect of endomorphins on the NMDA- and natural stimulus evoked responses. We suggest that endomorphins by acting at micro-opioid receptor selectively modulate noxious stimulus-evoked responses in the medullary dorsal horn. PMID- 10848573 TI - Human anterior cingulate cortex neurons modulated by attention-demanding tasks. AB - Recent imaging studies have implicated the anterior cingulate cortex (ACC) in various cognitive functions, including attention. However, until now, there was no evidence for changes in neuronal activity of individual ACC neurons during performance of tasks that require attention and effortful thought. We hypothesized these neurons must exist in the human ACC. In this study, we present electrophysiological data from microelectrode single neuron recordings in the human ACC of neuronal modulation during attention-demanding tasks in 19% of 36 neurons tested. These findings provide the first direct evidence of an influence of a cognitive state on the spontaneous neuronal activity of human ACC neurons. PMID- 10848574 TI - Onset of cross-modal synthesis in the neonatal superior colliculus is gated by the development of cortical influences. AB - Many neurons in the superior colliculus (SC) are able to integrate combinations of visual, auditory, and somatosensory stimuli, thereby markedly affecting the vigor of their responses to external stimuli. However, this capacity for multisensory integration is not inborn. Rather, it appears comparatively late in postnatal development and is not expressed until the SC passes through several distinct developmental stages. As shown here, the final stage in this sequence is one in which a region of association cortex establishes functional control over the SC, thus enabling the multisensory integrative capabilities of its target SC neurons. The first example of this corticotectal input was seen at postnatal day 28. For any individual SC neuron, the onset of corticotectal influences appeared to be abrupt. Because this event occurred at very different times for different SC neurons, a period of 3-4 postnatal months was required before the adult-like condition was achieved. The protracted postnatal period required for the maturation of these corticotectal influences corresponded closely with estimates of the peak period of cortical plasticity, raising the possibility that the genesis of these corticotectal influences, and hence the onset of SC multisensory integration, occurs only after the cortex is capable of exerting experience dependent control over SC neurons. PMID- 10848575 TI - Testing cyclin specificity in the exit from mitosis. AB - Cyclical inactivation of B-type cyclins has been proposed to be required for alternating DNA replication and mitosis. Destruction box-dependent Clb5p degradation is strongly increased in mitotic cells, and constitutive overexpression of Clb5p lacking the destruction box resulted in rapid accumulation of inviable cells, frequently multiply budded, with DNA contents ranging from unreplicated to apparently fully replicated. Loss of viability correlated with retention of nuclear Clb5p at the time of nuclear division. CLB2 Deltadb overexpression that was quantitatively comparable to CLB5-Deltadb overexpression with respect to Clb protein production and Clb-associated kinase activity resulted in a distinct phenotype: reversible mitotic arrest with uniformly replicated DNA. Simultaneous overexpression of CLB2-Deltadb and CLB5 Deltadb overexpressers similarly resulted in a uniform arrest with replicated DNA, and this arrest was significantly more reversible than that observed with CLB5-Deltadb overexpression alone. These results suggest that Clb2p and not Clb5p can efficiently block mitotic completion. We speculate that CLB5-Deltadb overexpression may be lethal, because persistence of high nuclear Clb5p associated kinase throughout mitosis leads to failure to load origins of replication, thus preventing DNA replication in the succeeding cell cycle. PMID- 10848577 TI - Stat1 as a component of tumor necrosis factor alpha receptor 1-TRADD signaling complex to inhibit NF-kappaB activation. AB - Activated tumor necrosis factor alpha (TNF-alpha) receptor 1 (TNFR1) recruits TNFR1-associated death domain protein (TRADD), which in turn triggers two opposite signaling pathways leading to caspase activation for apoptosis induction and NF-kappaB activation for antiapoptosis gene upregulation. Here we show that Stat1 is involved in the TNFR1-TRADD signaling complex, as determined by employing a novel antibody array screening method. In HeLa cells, Stat1 was associated with TNFR1 and this association was increased with TNF-alpha treatment. TNFR1 signaling factors TRADD and Fas-associated death domain protein (FADD) were also found to interact with Stat1 in a TNF-alpha-dependent process. Our in vitro recombinant protein-protein interaction studies demonstrated that Stat1 could directly interact with TNFR1 and TRADD but not with FADD. Interaction between Stat1 and receptor-interacting protein (RIP) or TNFR-associated factor 2 (TRAF2) was not detected. Examination of Stat1-deficient cells showed an apparent increase in TNF-alpha-induced TRADD-RIP and TRADD-TRAF2 complex formation, while interaction between TRADD and FADD was unaffected. As a consequence, TNF-alpha mediated I-kappaB degradation and NF-kappaB activation were markedly enhanced in Stat1-deficient cells, whereas overexpression of Stat1 in 293T cells blocked NF kappaB activation by TNF-alpha. Thus, Stat1 acts as a TNFR1-signaling molecule to suppress NF-kappaB activation. PMID- 10848576 TI - Mapping of atypical protein kinase C within the nerve growth factor signaling cascade: relationship to differentiation and survival of PC12 cells. AB - The pathway by which atypical protein kinase C (aPKC) contributes to nerve growth factor (NGF) signaling is poorly understood. We previously reported that in PC12 cells NGF-induced activation of mitogen-activated protein kinase (MAPK) occurs independently of classical and nonclassical PKC isoforms, whereas aPKC isoforms were shown to be required for NGF-induced differentiation. NGF-induced activation of PKC-iota was observed to be dependent on phosphatidylinositol 3-kinase (PI3K) and led to coassociation of PKC-iota with Ras and Src. Expression of dominant negative mutants of either Src (DN2) or Ras (Asn-17) impaired activation of PKC iota by NGF. At the level of Raf-1, neither PKC-iota nor PI3 kinase was required for activation; however, PKC-iota could weakly activate MEK. Inhibitors of PKC iota activity and PI3K had no effect on NGF-induced MAPK or p38 activation but reduced NGF-stimulated c-Jun N-terminal kinase activity. Src, PI3K, and PKC-iota were likewise required for NGF-induced NF-kappaB activation and cell survival, whereas Ras was not required for either survival or NF-kappaB activation but was required for differentiation. IKK existed as a complex with PKC-iota, Src and IkappaB. Consistent with a role for Src in regulating NF-kappaB activation, an absence of Src activity impaired recruitment of PKC-iota into an IKK complex and markedly impaired NGF-induced translocation of p65/NF-kappaB to the nucleus. These findings reveal that in PC12 cells, aPKCs comprise a molecular switch to regulate differentiation and survival responses coupled downstream to NF-kappaB. On the basis of these findings, Src emerges as a critical upstream regulator of both PKC-iota and the NF-kappaB pathway. PMID- 10848578 TI - Sugar control of the plant cell cycle: differential regulation of Arabidopsis D type cyclin gene expression. AB - In most plants, sucrose is the major transported carbon source. Carbon source availability in the form of sucrose is likely to be a major determinant of cell division, and mechanisms must exist for sensing sugar levels and mediating appropriate control of the cell cycle. We show that sugar availability plays a major role during the G(1) phase by controlling the expression of CycD cyclins in Arabidopsis. CycD2 mRNA levels increase within 30 min of the addition of sucrose; CycD3 is induced after 4 h. This corresponds to induction of CycD2 expression early in G(1) and CycD3 expression in late G(1) near the S-phase boundary. CycD2 and CycD3 induction is independent both of progression to a specific point in the cell cycle and of protein synthesis. Protein kinase activity of CycD2- and CycD3 containing cyclin-dependent kinases is consistent with the observed regulation of their mRNA levels. CycD2 and CycD3 therefore act as direct mediators of the presence of sugar in cell cycle commitment. CycD3, but not CycD2, expression responds to hormones, for which we show that the presence of sugars is required. Finally, protein phosphatases are shown to be involved in regulating CycD2 and CycD3 induction. We propose that control of CycD2 and CycD3 by sucrose forms part of cell cycle control in response to cellular carbohydrate status. PMID- 10848579 TI - Processing of intron-encoded box C/D small nucleolar RNAs lacking a 5',3' terminal stem structure. AB - The C and D box-containing (box C/D) small nucleolar RNAs (snoRNAs) function in the nucleolytic processing and 2'-O-methylation of precursor rRNA. In vertebrates, most box C/D snoRNAs are processed from debranched pre-mRNA introns by exonucleolytic activities. Elements directing accurate snoRNA excision are located within the snoRNA itself; they comprise the conserved C and D boxes and an adjoining 5',3'-terminal stem. Although the terminal stem has been demonstrated to be essential for snoRNA accumulation, many snoRNAs lack a terminal helix. To identify the cis-acting elements supporting the accumulation of intron-encoded box C/D snoRNAs devoid of a terminal stem, we have investigated the in vivo processing of the human U46 snoRNA and an artificial snoRNA from the human beta-globin pre-mRNA. We demonstrate that internal and/or external stem structures located within the snoRNA or in the intronic flanking sequences support the accumulation of mammalian box C/D snoRNAs lacking a canonical terminal stem. In the intronic precursor RNA, transiently formed external and/or stable internal base-pairing interactions fold the C and D boxes together and therefore facilitate the binding of snoRNP proteins. Since the external intronic stems are degraded during snoRNA processing, we propose that the C and D boxes alone can provide metabolic stability for the mature snoRNA. PMID- 10848580 TI - PKR stimulates NF-kappaB irrespective of its kinase function by interacting with the IkappaB kinase complex. AB - The interferon (IFN)-induced double-stranded RNA-activated protein kinase PKR mediates inhibition of protein synthesis through phosphorylation of the alpha subunit of eukaryotic initiation factor 2 (eIF2alpha) and is also involved in the induction of the IFN gene through the activation of the transcription factor NF kappaB. NF-kappaB is retained in the cytoplasm through binding to its inhibitor IkappaBalpha. The critical step in NF-kappaB activation is the phosphorylation of IkappaBalpha by the IkappaB kinase (IKK) complex. This activity releases NF kappaB from IkappaBalpha and allows its translocation to the nucleus. Here, we have studied the ability of PKR to activate NF-kappaB in a reporter assay and have shown for the first time that two catalytically inactive PKR mutants, PKR/KR296 and a deletion mutant (PKR/Del42) which lacks the potential eIF2alpha binding domain, can also activate NF-kappaB. This result indicated that NF-kappaB activation by PKR does not require its kinase activity and that it is independent of the PKR-eIF2alpha relationship. Transfection of either wild-type PKR or catalytically inactive PKR in PKR(0/0) mouse embryo fibroblasts resulted in the activation of the IKK complex. By using a glutathione S-transferase pull-down assay, we showed that PKR interacts with the IKKbeta subunit of the IKK complex. This interaction apparently does not require the integrity of the IKK complex, as it was found to occur with extracts from cells deficient in the NF-kappaB essential modulator, one of the components of the IKK complex. Therefore, our results reveal a novel pathway by which PKR can modulate the NF-kappaB signaling pathway without using its kinase activity. PMID- 10848581 TI - Involvement of the MKK6-p38gamma cascade in gamma-radiation-induced cell cycle arrest. AB - The p38 group of kinases belongs to the mitogen-activated protein (MAP) kinase superfamily with structural and functional characteristics distinguishable from those of the ERK, JNK (SAPK), and BMK (ERK5) kinases. Although there is a high degree of similarity among members of the p38 group in terms of structure and activation, each member appears to have a unique function. Here we show that activation of p38gamma (also known as ERK6 or SAPK3), but not the other p38 isoforms, is required for gamma-irradiation-induced G(2) arrest. Activation of the MKK6-p38gamma cascade is sufficient to induce G(2) arrest in cells, and expression of dominant negative alleles of MKK6 or p38gamma allows cells to escape the DNA damage-induce G(2) delay. Activation of p38gamma is dependent on ATM and leads to activation of Cds1 (also known as Chk2). These data suggest a model in which activation of ATM by gamma irradiation leads to the activation of MKK6, p38gamma, and Cds1 and that activation of both MKK6 and p38gamma is essential for the proper regulation of the G(2) checkpoint in mammalian cells. PMID- 10848582 TI - Disruption of mouse SNM1 causes increased sensitivity to the DNA interstrand cross-linking agent mitomycin C. AB - DNA interstrand cross-links (ICLs) represent lethal DNA damage, because they block transcription, replication, and segregation of DNA. Because of their genotoxicity, agents inducing ICLs are often used in antitumor therapy. The repair of ICLs is complex and involves proteins belonging to nucleotide excision, recombination, and translesion DNA repair pathways in Escherichia coli, Saccharomyces cerevisiae, and mammals. We cloned and analyzed mammalian homologs of the S. cerevisiae gene SNM1 (PSO2), which is specifically involved in ICL repair. Human Snm1, a nuclear protein, was ubiquitously expressed at a very low level. We generated mouse SNM1(-/-) embryonic stem cells and showed that these cells were sensitive to mitomycin C. In contrast to S. cerevisiae snm1 mutants, they were not significantly sensitive to other ICL agents, probably due to redundancy in mammalian ICL repair and the existence of other SNM1 homologs. The sensitivity to mitomycin C was complemented by transfection of the human SNM1 cDNA and by targeting of a genomic cDNA-murine SNM1 fusion construct to the disrupted locus. We also generated mice deficient for murine SNM1. They were viable and fertile and showed no major abnormalities. However, they were sensitive to mitomycin C. The ICL sensitivity of the mammalian SNM1 mutant suggests that SNM1 function and, by implication, ICL repair are at least partially conserved between S. cerevisiae and mammals. PMID- 10848583 TI - RanGTP-binding protein NXT1 facilitates nuclear export of different classes of RNA in vitro. AB - To better characterize the mechanisms responsible for RNA export from the nucleus, we developed an in vitro assay based on the use of permeabilized HeLa cells. This new assay supports nuclear export of U1 snRNA, tRNA, and mRNA in an energy- and Xenopus extract-dependent manner. U1 snRNA export requires a 5' monomethylated cap structure, the nuclear export signal receptor CRM1, and the small GTPase Ran. In contrast, mRNA export does not require the participation of CRM1. We show here that NXT1, an NTF2-related protein that binds directly to RanGTP, strongly stimulates export of U1 snRNA, tRNA, and mRNA. The ability of NXT1 to promote export is dependent on its capacity to bind RanGTP. These results support the emerging view that NXT1 is a general export factor, functioning on both CRM1-dependent and CRM1-independent pathways of RNA export. PMID- 10848584 TI - A GG nucleotide sequence of the 3' untranslated region of amyloid precursor protein mRNA plays a key role in the regulation of translation and the binding of proteins. AB - The alternative polyadenylation of the mRNA encoding the amyloid precursor protein (APP) involved in Alzheimer's disease generates two molecules, with the first of these containing 258 additional nucleotides in the 3' untranslated region (3'UTR). We have previously shown that these 258 nucleotides increase the translation of APP mRNA injected in Xenopus oocytes (5). Here, we demonstrate that this mechanism occurs in CHO cells as well. We also present evidence that the 3'UTR containing 8 nucleotides more than the short 3'UTR allows the recovery of an efficiency of translation similar to that of the long 3'UTR. Moreover, the two guanine residues located at the 3' ends of these 8 nucleotides play a key role in the translational control. Using gel retardation mobility shift assay, we show that proteins from Xenopus oocytes, CHO cells, and human brain specifically bind to the short 3'UTR but not to the long one. The two guanine residues involved in the translational control inhibit this specific binding by 65%. These results indicate that there is a correlation between the binding of proteins to the 3'UTR of APP mRNA and the efficiency of mRNA translation, and that a GG motif controls both binding of proteins and translation. PMID- 10848585 TI - Involvement of p21(Waf1/Cip1) in protein kinase C alpha-induced cell cycle progression. AB - Protein kinase C (PKC) plays an important role in the regulation of glioma growth; however, the identity of the specific isoform and mechanism by which PKC fulfills this function remain unknown. In this study, we demonstrate that PKC activation in glioma cells increased their progression through the cell cycle. Of the six PKC isoforms that were present in glioma cells, PKC alpha was both necessary and sufficient to promote cell cycle progression when stimulated with phorbol 12-myristate 13-acetate. Also, decreased PKC alpha expression resulted in a marked decrease in cell proliferation. The only cell cycle-regulatory molecule whose expression was rapidly altered and increased by PKC alpha activity was the cyclin-cyclin-dependent kinase (CDK) inhibitor p21(Waf1/Cip1). Coimmunoprecipitation studies revealed that p21(Waf1/Cip1) upregulation was accompanied by an incorporation of p21(Waf1/Cip1) into various cyclin-CDK complexes and that the kinase activity of these complexes was increased, thus resulting in cell cycle progression. Furthermore, depletion of p21(Waf1/Cip1) by antisense strategy attenuated the PKC-induced cell cycle progression. These results suggest that PKC alpha activity controls glioma cell cycle progression through the upregulation of p21(Waf1/Cip1), which facilitates active cyclin-CDK complex formation. PMID- 10848586 TI - Upf1p control of nonsense mRNA translation is regulated by Nmd2p and Upf3p. AB - Upf1p, Nmd2p, and Upf3p regulate the degradation of yeast mRNAs that contain premature translation termination codons. These proteins also appear to regulate the fidelity of termination, allowing translational suppression in their absence. Here, we have devised a novel quantitative assay for translational suppression, based on a nonsense allele of the CAN1 gene (can1-100), and used it to determine the regulatory roles of the UPF/NMD gene products. Deletion of UPF1, NMD2, or UPF3 stabilized the can1-100 transcript and promoted can1-100 nonsense suppression. Changes in mRNA levels were not the basis of suppression, however, since deletion of DCP1 or XRN1 or high-copy-number can1-100 expression in wild type cells caused an increase in mRNA abundance similar to that obtained in upf/nmd cells but did not result in comparable suppression. can1-100 suppression was highest in cells harboring a deletion of UPF1, and overexpression of UPF1 in cells with individual or multiple upf/nmd mutations lowered the level of nonsense suppression without affecting the abundance of the can1-100 mRNA. Our findings indicate that Nmd2p and Upf3p regulate Upf1p activity and that Upf1p plays a critical role in promoting termination fidelity that is independent of its role in regulating mRNA decay. Consistent with these relationships, Upf1p, Nmd2p, and Upf3p were shown to be present at 1, 600, 160, and 80 molecules per cell, levels that underscored the importance of Upf1p but minimized the likelihood that these proteins were associated with all ribosomes or that they functioned as a stoichiometric complex. PMID- 10848587 TI - Eap1p, a novel eukaryotic translation initiation factor 4E-associated protein in Saccharomyces cerevisiae. AB - Ribosome binding to eukaryotic mRNA is a multistep process which is mediated by the cap structure [m(7)G(5')ppp(5')N, where N is any nucleotide] present at the 5' termini of all cellular (with the exception of organellar) mRNAs. The heterotrimeric complex, eukaryotic initiation factor 4F (eIF4F), interacts directly with the cap structure via the eIF4E subunit and functions to assemble a ribosomal initiation complex on the mRNA. In mammalian cells, eIF4E activity is regulated in part by three related translational repressors (4E-BPs), which bind to eIF4E directly and preclude the assembly of eIF4F. No structural counterpart to 4E-BPs exists in the budding yeast, Saccharomyces cerevisiae. However, a functional homolog (named p20) has been described which blocks cap-dependent translation by a mechanism analogous to that of 4E-BPs. We report here on the characterization of a novel yeast eIF4E-associated protein (Eap1p) which can also regulate translation through binding to eIF4E. Eap1p shares limited homology to p20 in a region which contains the canonical eIF4E-binding motif. Deletion of this domain or point mutation abolishes the interaction of Eap1p with eIF4E. Eap1p competes with eIF4G (the large subunit of the cap-binding complex, eIF4F) and p20 for binding to eIF4E in vivo and inhibits cap-dependent translation in vitro. Targeted disruption of the EAP1 gene results in a temperature-sensitive phenotype and also confers partial resistance to growth inhibition by rapamycin. These data indicate that Eap1p plays a role in cell growth and implicates this protein in the TOR signaling cascade of S. cerevisiae. PMID- 10848588 TI - Hsl1p, a Swe1p inhibitor, is degraded via the anaphase-promoting complex. AB - Ubiquitination and subsequent degradation of critical cell cycle regulators is a key mechanism exploited by the cell to ensure an irreversible progression of cell cycle events. The anaphase-promoting complex (APC) is a ubiquitin ligase that targets proteins for degradation by the 26S proteasome. Here we identify the Hsl1p protein kinase as an APC substrate that interacts with Cdc20p and Cdh1p, proteins that mediate APC ubiquitination of protein substrates. Hsl1p is absent in G(1), accumulates as cells begin to bud, and disappears in late mitosis. Hsl1p is stabilized by mutations in CDH1 and CDC23, both of which result in compromised APC activity. Unlike Hsl1p, Gin4p and Kcc4p, protein kinases that have sequence homology to Hsl1p, were stable in G(1)-arrested cells containing active APC. Mutation of a destruction box motif within Hsl1p (Hsl1p(db-mut)) stabilized Hsl1p. Interestingly, this mutation also disrupted the Hsl1p-Cdc20p interaction and reduced the association between Hsl1p and Cdh1p in coimmunoprecipitation studies. These findings suggest that the destruction box motif is required for Cdc20p and, to a lesser extent, for Cdh1p to target Hsl1p to the APC for ubiquitination. Hsl1p has been previously shown to inhibit Swe1p, a protein kinase that negatively regulates the cyclin-dependent kinase Cdc28p, by promoting Swe1p degradation via SCF(Met30) in a bud morphogenesis checkpoint. Results of the present work indicate that Hsl1p is degraded in an APC-dependent manner and suggest a link between the SCF (Skp1-cullin-F box) and APC-proteolytic systems that may help to coordinate the proper progression of cell cycle events. PMID- 10848589 TI - Estrogen opposes the apoptotic effects of bone morphogenetic protein 7 on tissue remodeling. AB - Interactions between estrogen and growth factor signaling pathways at the level of gene expression play important roles in the function of reproductive tissues. For example, estrogen regulates transforming growth factor beta (TGFbeta) in the uterus during the proliferative phase of the mammalian reproductive cycle. Bone morphogenetic protein 7 (BMP-7), a member of the TGFbeta superfamily, is also involved in the development and function of reproductive tissues. However, relatively few studies have addressed the expression of BMP-7 in reproductive tissues, and the role of BMP-7 remains unclear. As part of an ongoing effort to understand how estrogen represses gene expression and to study its interactions with other signaling pathways, chick BMP-7 (cBMP-7) was cloned. cBMP-7 mRNA levels are repressed threefold within 8 h following estrogen treatment in the chick oviduct, an extremely estrogen-responsive reproductive tissue. This regulation occurs at the transcriptional level. Estrogen has a protective role in many tissues, and withdrawal from estrogen often leads to tissue regression; however, the mechanisms mediating regression of the oviduct remain unknown. Terminal transferase-mediated end-labeling and DNA laddering assays demonstrated that regression of the oviduct during estrogen withdrawal involves apoptosis, which is a novel observation. cBMP-7 mRNA levels during estrogen withdrawal increase concurrently with the apoptotic index of the oviduct. Furthermore, addition of purified BMP-7 induces apoptosis in primary oviduct cells. This report demonstrates that the function of BMP-7 in the oviduct involves the induction of apoptosis and that estrogen plays an important role in opposing this function.